PMID- 9846616 TI - Systematic relationships of some members of the genera Oesophagostomum and Chabertia (Nematoda: Chabertiidae) based on ribosomal DNA sequence data. AB - The present study characterised seven species of the Chabertiidae (Nematoda: Strongyloidea) belonging to either the subfamily Oesophagostominae (Oesophagostomum radiatum, Oesophagostomum venulosum, Oesophagostomum dentatum, Oesophagostomum quadrispinulatum, Oesophagostomum columbianum, Oesophagostomum bifurcum) or to the subfamily Chabertiinae (Chabertia ovina) by their second internal transcribed spacer rDNA sequence, assessed the extent of intraspecific variation and interspecific differences in the sequence, and inferred the phylogenetic relationship of C. ovina with respect to members of the Oesophagostominae. In both the phenetic and cladistic analyses of the sequence data, Chabertia was nested within Oesophagostomum, suggesting either that the species examined represent members of the same genus, or alternatively, that Oesophagostomum may represent more than one genus. PMID- 9846617 TI - A DNA-based demonstration of a three-host life-cycle for the Bivesiculidae (Platyhelminthes: Digenea) AB - Immature bivesiculid trematodes collected from the intestine of Thlalassoma lunare (Labridae) are shown to be morphologically consistent with adults of Bivesicula claviformis from Epinephelus fasciatus (Serranidae). In addition, the immature bivesiculids have the same sequence for the second internal transcribed spacer of the ribosomal DNA. Comparison with three other species of Bivesiculidae showed differences of between 23% and 30%. These results show that bivesiculids may have three-host life-cycles in addition to the two-host life-cycles that have been demonstrated previously. The three-host life-cycle enables bivesiculids to infect large carnivorous fishes. PMID- 9846618 TI - Genetic resistance to gastrointestinal nematode parasites in Polish long-wool sheep. AB - A study was undertaken from 1991 to 1994 on a farm in southern Poland to evaluate the genetic parameters of resistance to gastrointestinal nematodes. The predominant species were Teladorsagia circumcincta and Haemonchus contortus. A total of 32 sires were evaluated, around 15 per year. Faecal egg counts were measured twice during the 4-month grazing season for lambs (total 659 lambs) and three times for their mothers (total 327 ewes). Infection levels were high during the first 2 years and low during the last 2 years. Using an animal model, the heritability of log10(epg+25) increased from 0.20 in August to 0.33 in September for lambs, and from 0.18 in May to 0.25 in September for ewes. The repeatability of ewe faecal egg count between years was 0.25. A genetic correlation of 0.58 was found between faecal egg count in ewes and in 6-7-month-old lambs. A negative genetic correlation (-0.61) was estimated between faecal egg count in September and daily weight gain of lambs from 70 days of age to the end of grazing season (7 months of age). The results confirm the feasibility of genetic selection of sheep for resistance to nematode parasites in an environment where T. circumcincta and H. contortus are the dominant species. PMID- 9846619 TI - The uterine epithelium of Gyrodactylus kobayashii (Monogenea: Gyrodactylidae): ultrastructure of basal matrices, cytoplasmic membranes and the birth plug, and comparison with other reproductive epithelia. AB - Ultrastructural details of reproductive epithelia in the viviparous monogenean Gyrodactylus kobayashii are described. Specimens of G. kobayashii were fixed for transmission electron microscopy in glutaraldehyde in sodium cacodylate buffer followed by either 1% aqueous osmium tetroxide or 1% aqueous osmium tetroxide reduced with 1.5% potassium ferricyanide. All reproductive epithelia are underlain by a fibrillar basal matrix. The uterine basal matrix is electron opaque after potassium ferricyanide reduced osmium tetroxide fixation suggesting the presence of carbohydrate-containing materials. With potassium ferricyanide reduced osmium tetroxide fixation, two prominent membrane systems were distinguished in the uterine epithelium. Basal invaginations are short infoldings of the basal membrane. The basal invaginations are common in other reproductive epithelia and tegument and probably enhance transport of materials by these epithelia. Laminated membranes are membrane stacks, resembling endoplasmic reticulum stacks. These membranes were abundant at the apical membrane. The birth plug is a solid cytoplasmic layer, lacking a lumen, and rich in cytoplasmic vesicles. This layer connects the tegument and the uterine epithelium. PMID- 9846620 TI - Evolution of parasites' fecundity. PMID- 9846621 TI - Lp82 calpain during rat lens maturation and cataract formation. AB - PURPOSE: To measure changes in levels of Lp82 during maturation and selenite cataract formation in rat lens. Lp82 is a lens-specific, calcium-activated isozyme from the calpain family of cysteine proteases (EC 34.22.17). METHODS: Competitive RT-PCR was used to assess Lp82 and m-calpain mRNA concentrations. Immunoblotting and ELISA after DEAE chromatography measured Lp82 and m-calpain protein levels. Casein zymography assessed proteolytic activities in regions and whole lenses from maturing rats. RESULTS: Levels of Lp82 mRNA, protein, and caseinolytic activity decreased more rapidly during maturation of rat lens than for m-calpain. Unexpectedly, the water-insoluble fraction of rat lens contained enzymatically active Lp82. Selenite injection also caused major loss of Lp82 protein during cataract formation. CONCLUSIONS: Lp82 is a proteolytic enzyme likely functioning in early lens development and maturation. The rapid loss of Lp82 activity during lens maturation is probably caused by three factors: autodegradation associated with the proteolysis of soluble and insoluble proteins occurring in the rat lens nucleus, association of Lp82 with the lens insoluble fraction, and loss of Lp82 mRNA. Lp82 may function early in lens maturation along with m-calpain, which then is predominant in the latter stages of maturation. Proteolysis in selenite cataract is partially caused by over-activation of Lp82. PMID- 9846622 TI - Comparison of rabbit and human corneas stored in Optisol-GS: changes in stromal sodium. AB - PURPOSE: To evaluate the hydration, and the levels of free, total and bound sodium in fresh rabbit corneal stromas and also those preserved for up to 21 days in Optisol-GS. The effect of epithelial removal on stromal sodium and hydration parameters was also evaluated. Trends in stromal hydration and sodium environment were compared to results we previously obtained using human eyes stored under identical conditions. METHODS: Stromal hydration was evaluated thermogravimetrically. A sodium-specific electrode and an atomic absorption spectrophotometer were used to determine the amounts of free and total stromal sodium, respectively. In one cornea of each pair, the epithelium was removed prior to placement in the storage media. After 3, 7, 14 or 21 days at 4 degrees C, corneas were removed from the Optisol-GS, at which time sodium and hydration measurements were obtained. RESULTS: With an intact epithelium, the hydration of the rabbit stromas was elevated significantly at each day of storage compared to fresh corneas. Free and total sodium levels of rabbit stromas did not differ statistically from fresh values, however the bound sodium values did increase during storage. In the absence of the epithelium, the stromal hydration and sodium content (free, total and bound) were significantly elevated and the increase was much greater than in corneas stored with an intact epithelium. These findings differ from those we measured previously using human tissue. CONCLUSIONS: Rabbit corneas responded differently from human corneas to storage in Optisol-GS. The hydration levels increased to a greater level in rabbit than human corneas under both storage conditions. The trends in amounts of both free and total sodium were similar between the species, although the absolute amounts differed. The largest discrepancy was observed in the amount of bound sodium, with the rabbit corneas experiencing large increases not documented in the human tissue. These results suggest that direct comparisons of stromal hydration and ionic environment between the species should be approached with caution. PMID- 9846623 TI - Ultrastructural examination of corneal epithelium of spontaneously obese, hyperglycemic rats. AB - PURPOSE: Otsuka Long-Evans Tokushima fatty (OLETF) rats spontaneously become obese and hyperglycemic with age. We investigated whether the development of hyperglycemia would alter the ultrastructure of the corneal epithelium. METHODS: Scanning and transmission electron microscopy (SEM and TEM) were used to examine the morphology of corneal epithelial cells. Fourteen OLETF rats were evaluated, and 9 Long-Evans Tokushima Otsuka (LETO) rats were used as control. Non hyperglycemic OLETF rats served as controls. RESULTS: SEM showed exfoliative changes in the surface of the central corneal epithelium of the hyperglycemic OLETF rats. These superficial epithelial cells were irregular in shape as compared to polygonal shapes of those of LETO and non-hyperglycemic OLETF rats. The mean anterior surface area of individual superficial epithelial cells was significantly smaller in the hyperglycemic OLETF than that of the LETO or the non hyperglycemic OLETF rats. Central protrusion(s) could be found in some of the superficial cells of all rats examined, although this phenomenon was more common in the hyperglycemic rats than in the non-hyperglycemic rats. TEM revealed that there were numerous cytoplasmic vacuoles and wide intercellular spaces in the central corneal epithelium of the hyperglycemic OLETF rats, but not in the non hyperglycemic rats. CONCLUSIONS: The development of spontaneous hyperglycemia in OLETF rats alters the ultrastructure of the corneal epithelium. The alterations included abnormalities of the corneal epithelial surface observed by SEM and the presence of intracellular vacuoles and enlarged intercellular spaces detected by TEM. PMID- 9846624 TI - In vivo tear-film thickness determination and implications for tear-film stability. AB - PURPOSE: Previous measurements of tear-film thickness in vivo are limited and cannot be easily applied in a clinical setting. A novel technique to measure tear film thickness indirectly is introduced here, requiring only a slit lamp, video camera, and computer. A recent fluid mechanical theory relates tear-film thickness h, to the tear meniscus radius R, tear surface tension or, tear viscosity mu, and upper lid velocity U. This theory yields the result that h/R = 2.12 (microU/sigma)2/3. All parameters except h/R are taken as known physical constants, and R was measured for each subject, allowing the above equation to establish h. Tear-film breakup was also evaluated and correlated with tear-film thickness. METHODS: A clinical study was performed in which aqueous tear-film thickness was determined for 45 subjects, including 24 non-lens subjects, 15 hydrogel contact lens wearers, and 6 RGP lens wearers. R was measured by instilling fluorescein dye in the form of an eyedrop and videotaping the tear meniscus in profile. Tear-film breakup was videotaped through the ocular port of the slit lamp and evaluated based on a severity scale. RESULTS: Aqueous tear-film measurements are in the same range as literature values, with most measured values falling between 6 and 12 microm. Average tear-film thicknesses for non lens, hydrogel, and RGP subjects are 10.4, 6.5, and 5.8 microm, respectively. Tear-film breakup is most severe in subjects with thin tear films, especially in contact-lens wearers. CONCLUSIONS: Tear-film thickness is an important parameter that varies among individuals. These variations correlate with differences in tear-film stability. PMID- 9846625 TI - Superoxide dismutases in retinal degeneration of WBN/Kob rat. AB - PURPOSE: To examine the relationship between retinal degeneration and superoxide dismutase (SOD) in the degenerative retina of the WBN/Kob rat. METHODS: The retinas of 4-week-old and 10-month-old WBN/ Kob rats were examined with immunohistochemistry and immunoblotting. Wistar Kyoto rats were used as controls. RESULTS: Retinal degeneration began 4 weeks after birth. Four-week-old WBN/Kob rats showed no manganese superoxide dismutase (Mn-SOD) immunoreactivity in the photoreceptor inner segments or copper-zinc superoxide dismutase (CuZn-SOD) immunoreactivity in the outer nuclear layer or photoreceptor inner segments. Control 4-week-old Wistar Kyoto rats showed positive immunoreactivities at these sites. CONCLUSIONS: The retinal degeneration of WBN/Kob rats begins in the outer retina. The lack of SODs in the outer retina may contribute to retinal degeneration in the WBN/Kob rats. PMID- 9846626 TI - Purification of gamma-crystallin from human lenses by acetone precipitation method. AB - PURPOSE: The aim of this study was to develop a new purification method for human lens gamma-crystallin by utilizing its unique property of remaining soluble during acetone precipitation of water soluble (WS) proteins. METHODS: The WS protein fractions from lenses of donors of different ages were precipitated with 50% acetone (v/v) and the supernatant and precipitated protein fractions were collected following centrifugation. Among lens crystallins, gamma-crystallin remained soluble (recovered in the supernatant following centrifugation) while other crystallins were precipitated. To determine the recovery of maximal levels of gamma-crystallin as soluble protein during acetone precipitation, the WS proteins were precipitated under different conditions, and both supernatant and precipitated fractions were quantified for proteins and analyzed by size exclusion chromatographic and Western blot methods. Based on these results, a three-step purification procedure for gamma-crystallin was developed which consisted of acetone precipitation followed by preparative isoelectric focusing (IEF) and size-exclusion HPLC of the soluble fraction. RESULTS: During precipitation of WS proteins by 50% (v/v) acetone, only gamma-crystallin remained soluble. The identity of gamma-crystallin was based on its Mr of 20 kDa on SDS PAGE, co-elution with lens homogenate gamma-crystallin during a size-exclusion Agarose chromatography, immunoreactivity with anti-gamma-crystallin antibody on a Western blot and an overlap of its partial N-terminal sequence with gammaC crystallin. A three-step procedure, as described above, provided a highly purified preparation of gammaC-crystallin from the WS protein fraction. The three step procedure was also utilized to recover a highly purified human lens recombinant gammaD-crystallin preparation from E. coli lysate. CONCLUSIONS: The unique property of human lens gamma-crystallin of remaining soluble during acetone precipitation can be utilized to purify this crystallin by a three-step procedure. This procedure is also applicable in the purification of recombinant gammaD-crystallin from E. coli lysate. PMID- 9846627 TI - Defensin gene expression in the cornea. AB - PURPOSE: To determine whether defensin genes are expressed in human corneas and bovine corneal keratocytes. METHODS: In situ hybridization and immunohistochemistry were used to localize defensin mRNA and protein in normal and diseased human corneas. Cultured bovine keratocytes were stimulated with IL 1alpha or TNFalpha to determine whether defensin mRNA production occurred. Reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify defensin cDNA from cytokine-induced keratocytes, and Southern blots were used to verify the specificity of RT-PCR amplification products. RESULTS: Defensin mRNA and protein were not detected in normal human corneal stroma, but were readily detectable in the corneal stroma in cases of rejected transplants and postinfectious keratitis. IL-1alpha was a potent inducer of defensin gene expression in keratocytes, which began 12 h after challenge and peaked at 18 to 24 h. TNFalpha weakly induced defensin mRNA in keratocytes at about 18 h. Southern blots of the RT-PCR products probed with an oligonucleotide complementary to internal sequences of defensin demonstrated the appropriately sized products (198 bp) specific for defensin. CONCLUSIONS: This report demonstrates the presence of defensin in the human cornea and the capacity of corneal keratocytes to produce defensin mRNA in response to IL-1alpha and TNFalpha. Release of defensins by keratocytes in response to cytokines elaborated in corneal inflammation may contribute to the host defense response in microbial keratitis. PMID- 9846628 TI - Expression of vascular endothelial growth factor and its receptor, KDR, following retinal ischemia-reperfusion injury in the rat. AB - PURPOSE: There is considerable evidence that vascular endothelial growth factor (VEGF) mediates ocular neovascularization in retinal vascular diseases. We investigated the time-dependent changes in the expression of VEGF and its receptor KDR/ Flk in a transient retinal ischemia-reperfusion injury model. METHODS: Transient retinal ischemia was induced by increasing the intraocular pressure in albino rats eyes for 45 min. In situ hybridization was used to identify the retinal cells synthesizing VEGF mRNA and KDR mRNA at various times following reperfusion. Immunohistochemical analysis was also carried out to detect VEGF immunoreactivity. RESULTS: In the control, non-ischemic retinas, signals for VEGF mRNA and KDR mRNA were observed in the cells of the ganglion cell layer. Immunoreactivity to VEGF was also found in the nerve fiber layer, the ganglion cell layer, and the retinal pigment epithelial (RPE) cell layer. Immediately and 6 h after reperfusion, VEGF and KDR mRNA expression was markedly decreased, but recovered by 24 h to the levels observed in normal retinas. Immunoreactivity for VEGF was also decreased immediately and 6 h after reperfusion, and was detected in the endothelial cells of the retinal vessels after 24 h. Immunoreactivity to VEGF recovered by 48 h after reperfusion. CONCLUSIONS: The hybridization pattern of VEGF and KDR mRNA in the ganglion cell layer strongly suggests that the ganglion cells are the major source of this growth factor. The decrease of VEGF mRNA, KDR/Flk mRNA and VEGF protein levels after ischemia and recovery after reperfusion suggest that transient hypoxia might mediate short-term down-regulation of VEGF and KDR mRNA. PMID- 9846629 TI - Glutathione S-transferase of goat lens: evidence for expression of only class mu isoenzymes. AB - PURPOSE: It has been suggested that glutathione S-transferases (GSTs) of the mammalian ocular lens detoxify various toxicants and cataractogens; therefore, it provides protection against cataractogenesis. In this study the GST isoenzymes of goat lens have been purified and characterized. METHODS: The goat lens homogenate was subjected to affinity chromatography over GSH-linked epoxy activated Sepharose 6B. The isoenzymes of the GST were separated by chromatofocusing on a Mono-P column using FPLC. RESULTS: Two GST isoenzymes, GST 7.2 and GST 6.6, were purified to apparent homogeneity from goat lens. GST 7.2 appeared to be a heterodimer of Mr. 26,500 and Mr. 25,000 subunits, the GST 6.6 showed a homodimeric subunit structure of Mr. 25,000. Western blotting and the N-terminal region amino acid sequence analysis indicated that both isoenzymes of goat lens belong to the mu-class GST. CONCLUSION: Although they share varying degrees of structural correlation, the two isoenzymes of goat lens seem to be the products of two distinct genes. The isoenzyme expression pattern of GST in goat lens is similar to bovine lens, which also contains two isoenzymes belonging only to GST mu. PMID- 9846630 TI - Binding, gating, affinity and efficacy: the interpretation of structure-activity relationships for agonists and of the effects of mutating receptors. PMID- 9846631 TI - Evidence that [Phe1 psi(CH2-NH)Gly2]nociceptin-(1-13)-NH2, a peripheral ORL-1 receptor antagonist, acts as an agonist in the rat spinal cord. AB - [Phe1 psi(CH2-NH)Gly2]nociceptin-(1-13)-NH2, a pseudopeptide analogue of nociceptin is an antagonist in peripheral assays. Here, using in vivo electrophysiological recordings of dorsal horn neurones, [Phe1 psi(CH2 NH)Gly2]nociceptin-(1-13)-NH2 appears to have agonist activity after spinal administration. The noxious evoked activity of the neurones was inhibited by [Phe1 psi(CH2-NH)Gly2]nociceptin-(1-13)-NH2, which was as potent as nociceptin itself. PMID- 9846632 TI - EB1089, a synthetic analogue of vitamin D, induces apoptosis in breast cancer cells in vivo and in vitro. AB - 1. Effects of the synthetic vitamin D analogue EB1089 on indices of apoptosis in cultured human breast cancer cells and in nitrosomethylurea-induced rat mammary tumours in vivo were investigated. 2. At a dose of 0.5 microg kg(-1) body weight, EB1089 caused significant inhibition of tumour progression over the 28 day treatment period in the absence of a significant increase in serum calcium concentration. Higher doses of EB1089 (1 and 2.5 microg kg(-1)) produced substantial regression of the experimental tumours which was accompanied by a striking change in the histological appearance of tumours consistent with induction of tumour cell death. 3. Fragmentation of genomic DNA is a characteristic feature of apoptosis. With the terminal transferase (TdT) assay, 3' DNA breaks indicative of DNA fragmentation were detected histochemically in mammary tumour cells from animals treated with EB1089 (2.5 microg kg(-1)) for 14 days. 4. Effects of the vitamin D analogue on induction of apoptosis were examined in vitro using the MCF-7 human breast cancer cell line. Using the TUNEL method, positive nuclear staining indicative of DNA fragmentation was detected in cells treated for 4 days with 10 nM EB1089. Apoptosis was also quantitated using a cell death ELISA which revealed a time and dose dependent induction of apoptosis by EB1089. 5. The effects of EB1089 on the expression of two oncoproteins which may regulate apoptosis, bcl-2 and bax were examined by Western analysis. In MCF-7 cell cultures treated with 1,25(OH)2D3 or EB1089 (1 x 10(-8) M), bcl-2 protein levels were decreased in a time-dependent manner relative to control levels. In contrast bax protein was not markedly regulated by these compounds. Densitometric analyses indicate that the vitamin D compounds lower the bcl-2/bax ratio favouring increased susceptibility of MCF-7 cells to undergo apoptosis. 6. These results suggest that the synthetic vitamin D analogue EB1089 may promote tumour regression by inducing active cell death. PMID- 9846633 TI - Characterization of the endothelin receptor subtype mediating epithelium-derived relaxant nitric oxide release from guinea-pig trachea. AB - 1. The endothelin (ET) receptor subtype that mediates niric oxide (NO)-dependent airway relaxation in tracheal tube preparations precontracted with carbachol and pretreated with indomethacin was investigated. The release of NO induced by ET from guinea-pig trachea using a recently developed porphyrinic microsensor was also measured. 2. ET-1 (1 pM-100 nM) contracted tracheal tube preparations pretreated with the NO-synthase inhibitor, L-NMMA, and relaxed, in an epithelium dependent manner, preparations pretreated with the inactive enantiomer D-NMMA. The effect of L-NMMA was reversed by L-Arg, but not by D-Arg. 3. The selective ET(B) receptor agonists, IRL 1620 or sarafotoxin S6c, both (1 pM-100 nM) contracted tracheal tube preparations in a similar manner either after treatment with D-NMMA or with L-NMMA. In the presence of the ET(A) receptor antagonist, FR139317 (10 microM), ET-1 administration resulted in a contraction that was similar after either L-NMMA or D-NMMA. In the presence of the ET(B) receptor antagonist, BQ788 (1 microM), ET-1 relaxed and contracted tracheas pretreated with D-NMMA and L-NMMA, respectively. 4. Exposure of tracheal segments to ET-1 (1 1000 nM) caused a concentration-dependent increase in NO release that was reduced by L-NMMA. IRL1620 (1 microM) did not cause any significant NO release. FR139317 (10 microM), but not, BQ788 (1 microM), inhibited the NO release induced by ET-1. 5. These results demonstrate that in the isolated guinea-pig trachea activation of ET(B) receptors results in a contractile response, whereas activation of ET(A) receptors cause both a contraction, and an epithelium-dependent relaxation that is mediated by NO release. PMID- 9846634 TI - Effects of propafenone and 5-hydroxy-propafenone on hKv1.5 channels. AB - 1. The goal of this study was to analyse the effects of propafenone and its major metabolite, 5-hydroxy-propafenone, on a human cardiac K+ channel (hKv1.5) stably expressed in Ltk- cells and using the whole-cell configuration of the patch-clamp technique. 2. Propafenone and 5-hydroxy-propafenone inhibited in a concentration dependent manner the hKv1.5 current with K(D) values of 4.4+/-0.3 microM and 9.2+/-1.6 microM, respectively. 3. Block induced by both drugs was voltage dependent consistent with a value of electrical distance (referenced to the cytoplasmic side) of 0.17+/-0.55 (n=10) and 0.16+/-0.81 (n=16). 4. The apparent association (k) and dissociation (l) rate constants for propafenone were (8.9+/ 0.9) x 10(6) M(-1) s(-1) and 39.5+/-4.2 s(-1), respectively. For 5-hydroxy propafenone these values averaged (2.3+/-0.3) x 10(6) M(-1) s(-1) and 21.4+/-3.1 s(-1), respectively. 5. Both drugs reduced the tail current amplitude recorded at -40 mV after 250 ms depolarizing pulses to +60 mV, and slowed the deactivation time course resulting in a 'crossover' phenomenon when the tail currents recorded under control conditions and in the presence of each drug were superimposed. 6. Both compounds induced a small but statistically significant use-dependent block when trains of depolarizations at frequencies between 0.5 and 3 Hz were applied. 7. These results indicate that propafenone and its metabolite block hKv1.5 channels in a concentration-, voltage-, time- and use-dependent manner and the concentrations needed to observe these effects are in the therapeutical range. PMID- 9846635 TI - Weaning-induced development of delta-opioid receptors in rat brain: differential effects of guanine nucleotides and sodium upon ligand-receptor recognition. AB - 1. We have previously shown that weaning at day 21 increases delta-opioid receptor binding in the brain at day 25, which might be due to stimulation of the development of a delta-opioid receptor subtype or activation of G-protein coupling processes. 2. We have addressed the possibility that weaning stimulates coupling of the delta-receptor by homogenate binding studies with four agonist and one antagonist radioligand in the presence of a GTP analogue and Na+ in brain tissue from weaned and non-weaned animals. 3. Saturation studies with three agonist ligands ([3H]-deltorphin I, [3H]-S-Atc-Ile(5,6)deltorphin I and [3H]-R Atc-Ile(5,6)deltorphin II) showed higher levels of maximal binding in brains from 25-day weaned than in brains from non-weaned rats. The magnitude of the effects of GMPPNP and Na+ in decreasing this binding was ligand dependent and in each case was significantly more marked in brains from weaned animals. GMPPNP and Na+ were completely without effect on Bmax for, [3H]-S-Atc-Ile(5,6)deltorphin I and [3H]-R-Atc-Ile(5,6)deltorphin II in brains from non-weaned rats. 4. [3H] Ile(5,6)deltorphin II and [3H]-naltrindole showed no differences in labelling between weaned and non-weaned groups and both groups responded similarly to the effects of GMPPNP and Na+ treatment. 5. GMPPNP and Na+ had small effects on binding affinity (K(D)) for some of the agonist radioligands which were similar in both weaned and non-weaned groups. 6. Weaning induced increases in binding of delta-receptors in 25-day rats can be explained in terms of the way delta-agonist radioligands recognize the receptor environment. PMID- 9846636 TI - Excitatory motor and electrical effects produced by tachykinins in the human and guinea-pig isolated ureter and guinea-pig renal pelvis. AB - 1. In isolated tissue experiments, neurokinin A (NKA) produced concentration dependent contraction of human and guinea-pig ureter (pD2 = 6.7 and 7.2, respectively); an effect greatly reduced (>80% inhibition) by the tachykinin NK2 receptor-selective antagonist MEN 11420 (0.1 microM). The tachykinin NK1 and NK3 receptor agonists septide and senktide, respectively, were ineffective. 2. Electrical field stimulation (EFS) of the guinea-pig isolated renal pelvis produced an inotropic response blocked by MEN 11420 (0.01-1 microM). In the same preparation MEN 11420 (0.1 microM) blocked (apparent pK(B) = 8.2) the potentiation of spontaneous motor activity produced by the NK2 receptor-selective agonist [betaAla8]NKA(4-10). 3. In sucrose-gap experiments, EFS evoked action potentials (APs) accompanied by phasic contractions of human and guinea-pig ureter, which were unaffected by tetrodotoxin or MEN 11420 (3 microM), but were blocked by nifedipine (1-10 microM). NKA (1-3 microM) produced a slow membrane depolarization with superimposed APs and a tonic contraction with superimposed phasic contractions. NKA prolonged the duration of EFS-evoked APs and potentiated the accompanying contractions. MEN 11420 completely prevented the responses to NKA in both the human and guinea-pig ureter. 4. Nifedipine (1-10 microM) suppressed the NKA-evoked APs and phasic contractions in both human and guinea pig ureter, and slightly reduced the membrane depolarization induced by NKA. A tonic-type contraction of the human ureter in response to NKA persisted in the presence of nifedipine. 5. In conclusion, tachykinins produce smooth muscle excitation in both human and guinea-pig ureter by stimulating receptors of the NK2 type only. NK2 receptor activation depolarizes the membrane to trigger the firing of APs from latent pacemakers. PMID- 9846637 TI - Pharmacodynamic interaction between phenytoin and sodium valproate changes seizure thresholds and pattern. AB - 1. In this study we used cortical stimulation to assess the effects of phenytoin (PHT), sodium valproate (VPA), and their interaction on total motor seizure and on the constituent elements of the seizure. 2. PHT (40 mg kg(-1)) was administered as an intravenous bolus infusion to animals receiving either a continuous infusion of VPA or saline. VPA plasma concentration was maintained at levels that produced no detectable anticonvulsant effect. 3. Analysis of ictal components (eyes closure, jerk, gasp, forelimb, clonus, and hindlimb tonus) and their durations revealed both qualitative and quantitative differences in drug effects. 4. The anticonvulsant effect is represented by the increase in the duration of the stimulation required to reach a given seizure threshold. PHT significantly increased the duration of the stimulation and of the motor seizure. This increase was greatly enhanced by VPA. In addition, ictal component analysis revealed that the combination of PHT and VPA causes the reduction of a specific seizure component (JERK). 5. Neither the free fraction of PHT nor the biophase equilibration kinetics changes in the presence of VPA. It is concluded that the synergism may be due to a pharmacodynamic rather than a pharmacokinetic interaction. PMID- 9846638 TI - The action of calcium channel blockers on recombinant L-type calcium channel alpha1-subunits. AB - 1. CHO cells expressing the alpha(1C-a) subunit (cardiac isoform) and the alpha(1C-b) subunit (vascular isoform) of the voltage-dependent L-type Ca2+ channel were used to investigate whether tissue selectivity of Ca2+ channel blockers could be related to different affinities for alpha1C isoforms. 2. Inward current evoked by the transfected alpha1 subunit was recorded by the patch-clamp technique in the whole-cell configuration. 3. Neutral dihydropyridines (nifedipine, nisoldipine, (+)-PN200-110) were more potent inhibitors of alpha(1C )b-subunit than of alpha(1C-a)-subunit. This difference was more marked at a holding potential of -100 mV than at -50 mV. SDZ 207-180 (an ionized dihydropyridine) exhibited the same potency on the two isoforms. 4. Pinaverium (ionized non-dihydropyridine derivative) was 2 and 4 fold more potent on alpha(1C a) than on alpha(1C-b) subunit at Vh of -100 mV and -50 mV, respectively. Effects of verapamil were identical on the two isoforms at both voltages. 5. [3H]-(+)-PN 200-110 binding experiments showed that neutral dihydropyridines had a higher affinity for the alpha(1C-b) than for the alpha(1C-a) subunit. SDZ 207-180 had the same affinity for the two isoforms and pinaverium had a higher affinity for the alpha(1C-a) subunit than for the alpha(1C-b) subunit. 6. These results indicate marked differences among Ca2+ channel blockers in their selectivity for the alpha(1C-a) and alpha(1C-b) subunits of the Ca2+ channel. PMID- 9846639 TI - Learning impairments induced by glutamate blockade using dizocilpine (MK-801) in monkeys. AB - 1. This study investigated the effects of dizocilpine (MK-801) on learning ability in a non-human primate. Acquisition and reversal learning of visual discrimination tasks and acquisition of visuo-spatial discrimination tasks were assessed in marmosets using the Wisconsin General Test Apparatus. Dizocilpine impaired acquisition of visuo-spatial (conditional) tasks requiring spatial responses to coloured objects, and perceptually difficult visual discrimination tasks in which stimulus objects are painted black. Dizocilpine did not, however, impair either acquisition or reversal of a simple visual discrimination task using easily discriminated, coloured objects. 2. Motor effects of dizocilpine treatment, which have been seen in other primates, were examined by observation of the marmosets in their home cages, using both an automated locomotor activity monitor and 'blind', subjective counting of the number of abnormal movements in a given time period. Locomotor activity, assessed using the automated monitor, was not significantly affected at any of the doses tested. Incoordination, assessed by human observation of abnormal movements, was significantly increased only at a dose of 30 microg kg(-1) i.m., which was twice the highest dose used to assess the effects of dizocilpine on cognition. 3. We have, therefore, found an effect of dizocilpine on acquisition and reversal of some types of cognitive task, at a dose which does not cause significant motor effects. This demonstration of a cognitive deficit associated with glutamatergic blockade in a primate may be useful in understanding the contribution of glutamatergic dysfunction to cognitive decline in neurodegenerative disease, especially Alzheimer's disease. PMID- 9846640 TI - Effect of endothelin-1 (1-31) on extracellular signal-regulated kinase and proliferation of human coronary artery smooth muscle cells. AB - 1. We have previously found that human chymase cleaves big endothelins (ETs) at the Tyr31-Gly32 bond and produces 31-amino acid ETs (1-31), without any further degradation products. In this study, we investigated the effect of synthetic ET-1 (1-31) on the proliferation of cultured human coronary artery smooth muscle cells (HCASMCs). 2. ET-1 (1-31) increased [3H]-thymidine incorporation and cell numbers to a similar extent as ET-1 at 100 nM. This ET-1 (1-31)-induced [3H]-thymidine uptake was not affected by phosphoramidon, an inhibitor of ET-converting enzyme. It was, however, inhibited by BQ123, an endothelin ET(A) receptor antagonist, but not by BQ788, an endothelin ET(B) receptor antagonist. 3. By using an in-gel kinase assay, we demonstrated that ET-1 (1-31) activated extracellular signal regulated kinase 1/2 (ERK1/2) in a concentration-dependent manner (100 pM to 1 microM) in HCASMCs. ET-1 (1-31)-induced ERK1/2 activation was inhibited by BQ123, but not by BQ788 and phosphoramidon. Inhibition of protein kinase C (PKC) and ERK kinase also caused a reduction of ET-1 (1-31)-induced ERK1/2 activation, whereas tyrosine kinase inhibition had little effect. 4. Gel-mobility shift analysis revealed that the ERK1/2 activation was followed by an increase in transcription factor activator protein-1 DNA binding activity in HCASMCs. 5. Our results strongly suggest that ET-1 (1-31) itself stimulates HCASMC proliferation probably through endothelin ET(A) or ET(A)-like receptors. The underlining mechanism of cell growth by ET-1 (1-31) may be explained in part by PKC-dependent ERK1/2 activation. Since human chymase has been proposed to play a role in atherosclerosis, ET-1 (1-31) may be one of the mediators. PMID- 9846641 TI - Dual pathway for angiotensin II formation in human internal mammary arteries. AB - 1. Angiotensin converting enzyme (ACE) is thought to be the main enzyme to convert antiotensin I to the vasoactive angiotensin II. Recently, in the human heart, it was found that the majority of angiotensin II formation was due to another enzyme, identified as human heart chymase. In the human vasculature however, the predominance of either ACE or non-ACE conversion of angiotensin I remains unclear. 2. To study the effects of ACE- and chymase-inhibition on angiotensin II formation in human arteries, segments of internal mammary arteries were obtained from 37 patients who underwent coronary bypass surgery. 3. Organ bath experiments showed that 100 microM captopril inhibited slightly the response to angiotensin I (pD2 from 7.09+/-0.11-6.79+/-0.10, P<0.001), while 100 microM captopril nearly abolished the response to [pro10] angiotensin I, a selective substrate for ACE, and the maximum contraction was reduced from 83+/-19%-23+/-17% of the control response (P=0.01). A significant decrease of the pD2 of angiotensin I similar to captopril was observed in the presence of 50 microM chymostatin (pD2 from 7.36+/-0.13-6.99+/-0.15, P<0.039), without influencing the maximum response. In the presence of both inhibitors, effects were much more pronounced than either inhibitor alone, and a 300 times higher dose was needed to yield a significant contraction response to angiotensin I. 4 These results indicate the presence of an ACE and a non-ACE angiontensin II forming pathway in human internal mammary arteries. PMID- 9846642 TI - Effects of chronic application of propranolol on beta-adrenergic signal transduction in heart ventricles from myopathic BIO TO2 and control hamsters. AB - 1. In human congestive heart failure beta-adrenoceptor antagonists improve exercise tolerance and cardiac contractility. These beneficial effects are thought to reflect an up-regulation of cardiac beta-adrenoceptors, involving mainly the beta1-subtype. In the present study we evaluated the functional contribution of beta-adrenoceptor subtypes to stimulation of adenylyl cyclase in an animal model of dilated cardiomyopathy, and compared the effects of treatment with propranolol on cardiac beta-adrenergic signal transduction in myopathic and control hamsters. 2. Cardiomyopathic BIO TO2 hamsters and BIO F1B controls aged 270 days were used. In the treatment study, hamsters received drinking water with or without propranolol 40 mg kg(-1) d(-1) for 4 weeks prior to sacrifice. Density and subtype distribution of beta-adrenoceptors were determined in radioligand binding studies. Functional contributions of beta-adrenoceptors were evaluated by subtype-selective stimulation of adenylyl cyclase. Cardiac G-protein content was determined by immunoblotting. 3. Compared to BIO F1B controls, myopathic hamsters showed increases in cardiac total beta- and beta2-adrenoceptor density, G(s alpha) and G(i alpha) content. In BIO TO2 ventricles, beta1-adrenoceptors were almost completely uncoupled from adenylyl cyclase stimulation despite an unchanged density. Treatment of hamsters with propranolol resulted in increased density of beta1-adrenoceptors in both strains, but had no effect on their functional efficacy. Moreover, beta2-adrenergic stimulation of adenylyl cyclase was even reduced in propranolol-treated animals, which could not be explained by changes in cardiac G-protein content. 4. Cardiomyopathic BIO TO2 hamsters showed functional uncoupling of cardiac beta1-adrenoceptors, which could not be normalized by propranolol and, therefore, is unlikely to be solely due to agonist dependent desensitization. The paradoxical reduction in beta2-adrenergic efficiency in propranolol-treated myopathic and control hamsters deserves further investigation. PMID- 9846643 TI - Nicotine administration stimulates the in vivo N-methyl-D-aspartate receptor/nitric oxide/cyclic GMP pathway in rat hippocampus through glutamate release. AB - 1. The in vivo effects of nicotine on the nitric oxide (NO) synthase/cyclic GMP pathway of the adult rat hippocampus have been investigated by monitoring the levels of extracellular cyclic GMP during microdialysis in conscious unrestrained animals. 2. Intraperitoneal (i.p.) administration of nicotine caused elevation of cyclic GMP levels which was prevented by mecamylamine. The effect of nicotine was abolished by local infusion of the NO synthase inhibitor N(G)-nitro-L-arginine (L NOARG) or by the soluble guanylyl cyclase blocker 1H-[1,2,4]oxadiazolo[4.3 a]quinoxaline-1-one (ODQ). 3. Local administration of the NMDA receptor antagonists cis-4-(phosphonomethyl)-2-piperidinecarboxylic acid (CGS19755) and dizocilpine (MK-801) inhibited by about 60% the nicotine-induced elevation of cyclic GMP. Nicotine was able to stimulate cyclic GMP outflow also when administered directly into the hippocampus; the effect was sensitive to mecamylamine, L-NOARG, ODQ or MK-801. 4. Nicotine, either administered i.p. or infused locally, produced augmentation of glutamate and aspartate extracellular levels, whereas the outflows of gamma-aminobutyric acid (GABA) and glycine remained unaffected. Following local administration of high concentrations of nicotine, animals displayed symptoms of mild excitation (sniffing, increased motor and exploratory activity) during the first 20-40 min of infusion, followed by wet dog shake episodes; these behavioural effects were prevented by mecamylamine or MK-801, but not by L-NOARG or by ODQ. 5. It is concluded that (a) nicotine stimulates the production of NO and cyclic GMP in the hippocampus; (b) this occurs, at least in part, through release of glutamate/aspartate and activation of NMDA receptors. Modulation of the NMDA receptor/NO synthase/cyclic GMP pathway may be involved in the cognitive activities of nicotine. PMID- 9846644 TI - Potentiation of adenosine A1 receptor-mediated inositol phospholipid hydrolysis by tyrosine kinase inhibitors in CHO cells. AB - 1. The effect of protein tyrosine kinase inhibitors on human adenosine A1 receptor-mediated [3H]-inositol phosphate ([3H]-IP) accumulation has been studied in transfected Chinese hamster ovary cells (CHO-A1) cells. 2. In agreement with our previous studies the selective adenosine A1 receptor agonist N6 cyclopentyladenosine (CPA) stimulated the accumulation of [3H]-IPs in CHO-A1 cells. Pre-treatment with the broad spectrum tyrosine kinase inhibitor genistein (100 microM; 30 min) potentiated the responses elicited by 1 microM (199+/-17% of control CPA response) and 10 microM CPA (234+/-15%). Similarly, tyrphostin A47 (100 microM) potentiated the accumulation of [3H]-IPs elicited by 1 microM CPA (280+/-32%). 3. Genistein (EC50 = 13.7+/-1.2 microM) and tyrphostin A47 (EC50 = 10.4+/-3.9 microM) potentiated the [3H]-IP response to 1 microM CPA in a concentration-dependent manner. 4. Pre-incubation with the inactive analogues of genistein and tyrphostin A47, daidzein (100 microM; 30 min) and tyrphostin A1 (100 microM; 30 min), respectively, had no significant effect on the accumulation of [3H]-IPs elicited by 1 microM CPA. 5. Genistein (100 microM) had no significant effect on the accumulation of [3H]-IPs produced by the endogenous thrombin receptor (1 u ml(-1); 100+/-10% of control response). In contrast, tyrphostin A47 produced a small augmentation of the thrombin [3H]-IP response (148+/-13%). 6. Genistein (100 microM) had no effect on the [3H]-IP response produced by activation of the endogenous Gq-protein coupled CCK(A) receptor with the sulphated C-terminal octapeptide of cholecystokinin (1 microM CCK-8; 96+/-6% of control). In contrast, tyrphostin A47 (100 microM) caused a small but significant increase in the response to 1 microM CCK-8 (113+/-3% of control). 7. The phosphatidylinositol 3-kinase inhibitor LY 294002 (30 microM) and the MAP kinase kinase inhibitor PD 98059 (50 microM) had no significant effect on the [3H]-IP responses produced by 1 microM CPA and 1 microM CCK-8. 8. These observations suggest that a tyrosine kinase-dependent pathway may be involved in the regulation of human adenosine A1 receptor mediated [3H]-IP responses in CHO A1 cells. PMID- 9846645 TI - On the inhibition of voltage activated calcium currents in rat cortical neurones by the neuroprotective agent 619C89. AB - 1. The lamotrigine analogue 619C89, utilised to reduce postischaemic and posttraumatic neuronal injury, has been shown to inhibit sodium channels and cloned N-type calcium channels. To verify whether this neuroprotective agent also blocked native calcium channels, we have tested its action in cortical pyramidal neurones, acutely isolated from the adult rat brain. 2. 619C89 inhibited more than 90% of the high voltage-activated calcium currents recorded in the whole cell configuration. The response was relatively slow in onset (30-60 s), recovered incompletely (96%), but showed no consistent desensitization. 3. This inhibitory effect was not selective for any calcium channel subtype, being largely unaffected by omega-conotoxin-GVIA, omega-agatoxin-IVA, omega-conotoxin MVIIC and dihydropyridine antagonists. 4. Saturating responses to 619C89 were detected for concentrations > or = 50 microM. Dose-response curves revealed that 619C89 have an approximately 8 microM binding site. 5. The effect of 619C89 was dependent on the divalent concentrations in that its potency was reduced on increase of the charge carrier up to 20 mM barium. Since the lamotrigine analogue shifted to the right the dose-dependence of the cadmium block, the 619C89 mediated inhibition of calcium currents seemed to rely on a direct interaction with the channel pore. Functional implications are discussed. PMID- 9846646 TI - Functional response of the rat kidney to inhibition of nitric oxide synthesis: role of cytochrome p450-derived arachidonate metabolites. AB - 1. We tested the hypothesis that nitric oxide (NO) exerts a tonic inhibitory influence on cytochrome P450 (CYP450)-dependent metabolism of arachidonic acid (AA). 2. N(omega)-nitro-L-Arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase (NOS), increased mean blood pressure (MBP), from 91+/-6 to 137+/-5 mmHg, renal vascular resistance (RVR), from 9.9+/-0.6 to 27.4+/-2.5 mmHg ml(-1) min(-1), and reduced renal blood flow (RBF), from 9.8+/-0.7 to 6.5+/-0.6 ml min( 1)) and GFR from 1.2+/-0.2 to 0.6+/-0.2 ml 100 g(-1) min(-1)) accompanied by diuresis (UV, 1.7+/-0.3 to 4.3+/-0.8 microl 100 g(-1) min (-1)), and natriuresis (U(Na)V, 0.36+/-0.04 to 1.25+/-0.032 micromol 100 g(-1) min(-1)). 3. 12, 12 dibromododec-enoic acid (DBDD), an inhibitor of omega hydroxylase, blunted L-NAME induced changes in MBP, RVR, UV and U(Na)V by 63+/-8, 70+/-5, 45+/-8 and 42+/-9%, respectively, and fully reversed the reduction in GFR by L-NAME. Clotrimazole, an inhibitor of the epoxygenase pathway of CYP450-dependent AA metabolism, was without effect. 4. BMS182874 (5-dimethylamino)-N-(3,4-dimethyl-5-isoxazolyl)-1 naphthalenesulfo namide), an endothelin (ET)A receptor antagonist, also blunted the increases in MBP and RVR and the diuresis/natriuresis elicited by L-NAME without affecting GFR. 5. Indomethacin blunted L-NAME-induced increases in RVR, UV and U(Na)V. BMS180291 (1S-(1alpha,2alpha,3alpha,4alpha)]-2-[[3-[4-[(++ +pentylamino)carbonyl]-2-oxazolyl]-7-oxabicyclo[2.2.1]hept-2-yl ]methyl]benzenepropanoic acid), an endoperoxide receptor antagonist, attenuated the pressor and renal haemodynamic but not the renal tubular effects of L-NAME. 6. In conclusion, the renal functional effects of the CYP450-derived mediator(s) expressed after inhibition of NOS with L-NAME were prevented by inhibiting either CYP450 omega hydroxylase or cyclooxygenase or by antagonizing either ET(A) or endoperoxide receptors. 20-hydroxyeicosatetraenoic acid (20-HETE) fulfils the salient properties of this mediator. PMID- 9846647 TI - The mechanism of the nitric oxide-mediated enhancement of tert-butylhydroperoxide induced DNA single strand breakage. AB - 1. Caffeine (Cf) enhances the DNA cleavage induced by tert-butylhydroperoxide (tB OOH) in U937 cells via a mechanism involving Ca2+-dependent mitochondrial formation of DNA-damaging species (Guidarelli et al., 1997b). Nitric oxide (NO) is not involved in this process since U937 cells do not express the constitutive nitric oxide synthase (cNOS). 2. Treatment with the NO donors S-nitroso-N-acetyl penicillamine (SNAP, 10 microM), or S-nitrosoglutathione (GSNO, 300 microM), however, potentiated the DNA strand scission induced by 200 microM tB-OOH. The DNA lesions generated by tB-OOH alone, or combined with SNAP, were repaired with superimposable kinetics and were insensitive to anti-oxidants and peroxynitrite scavengers but suppressed by iron chelators. 3. SNAP or GSNO did not cause mitochondrial Ca2+ accumulation but their enhancing effects on the tB-OOH-induced DNA strand scission were prevented by ruthenium red, an inhibitor of the calcium uniporter of mitochondria. Furthermore, the enhancing effects of both SNAP and GSNO were identical to and not additive with those promoted by the Ca2+ mobilizing agents Cf or ATP. 4. The SNAP- or GSNO-mediated enhancement of the tB OOH-induced DNA cleavage was abolished by the respiratory chain inhibitors rotenone and myxothiazol and was not apparent in respiration-deficient cells. 5. It is concluded that, in cells which do not express the enzyme cNOS, exogenous NO enhances the accumulation of DNA single strand breaks induced by tB-OOH via a mechanism involving inhibition of complex III. PMID- 9846648 TI - Examination of somatostatin involvement in the inhibitory action of GIP, GLP-1, amylin and adrenomedullin on gastric acid release using a new SRIF antagonist analogue. AB - 1. The effect of a new type 2 selective somatostatin (SRIF) receptor antagonist (DC-41-33) on somatostatin-induced inhibition of pentagastrin-stimulated gastric acid secretion in conscious, chronic gastric fistula equipped rats was studied. 2. Infused intravenously, DC-41-33 dose-dependently inhibits SRIF-induced inhibition of pentagastrin-stimulated gastric acid secretion with an IC50 of 31.6+/-1.2 nmol kg(-1) versus 10 nmol kg(-1) SRIF and blocks the inhibitory effects of SRIF when simultaneously co-infused. Its effectiveness provides additional evidence that SRIF-inhibition of gastric acid release is a SRIF type 2 receptor-mediated process. 3. DC-41-33 is able to completely reverse the inhibitory effect of glucose-dependent insulinotropic polypeptides, GIP and GIP (1-30)NH2, and glucagon-like polypeptide, GLP-1(7-36)NH2, on pentagastrin stimulated gastric acid secretion thus confirming that they exert these effects through stimulation of endogenous SRIF release. 4. DC-41-33 only partially blocks potent amylin and adrenomedullin-induced inhibition of gastric acid secretion, therefore suggesting that somatostatin may not function as a primary mediator in the action of these peptides. 5. Our results indicate that DC-41-33, is a potent in vivo inhibitor of exogenous and endogenous SRIF in rats. It represents a new class of SRIF analogues which should eventually provide excellent tools for further evaluating the many physiological roles of SRIF and its five receptor subtypes. PMID- 9846649 TI - Interaction of neuromuscular blocking drugs with recombinant human m1-m5 muscarinic receptors expressed in Chinese hamster ovary cells. AB - 1. Neuromuscular blocking drugs (NMBD's) are known to produce cardiovascular side effects manifesting as brady/tachycardias. In this study we have examined the interaction of a range of steroidal NMBD's with recombinant human m1-m5 muscarinic receptors expressed in Chinese hamster ovary cells. Our main hypothesis is that NMBD's may interact with m2 (cardiac) muscarinic receptors. 2. All binding studies were performed with cell membranes prepared from CHO m1-m5 cells in 1 ml volumes of 20 mM HEPES, 1 mM MgCl2 at pH 7.4 for 1 h. Muscarinic receptors were labelled with [3H]-NMS and displacement studies were performed with pancuronium, vecuronium, pipecuronium, rocuronium and gallamine. In addition a range of muscarinic receptor subtype selective reference compounds were included. In order to determine the nature of any interaction the effects of pancuronium, rocuronium and vecuronium on methacholine inhibition of forskolin stimulated cyclic AMP formation in CHO m2 cells was examined. Cyclic AMP formation was assessed in whole cells using a radioreceptor assay. All data are mean +/- s.e.mean (n > or = 5). 3. The binding of [3H]-NMS was dose-dependent and saturable in all cells tested. Bmax and Kd values in m1-m5 cells were 2242+/-75, 165+/-13, 1877+/-33, 458+/-30, 127+/-2 fmol mg(-1) protein and 0.11+/-0.02, 0.15+/-0.01, 0.12+/-0.01, 0.12+/-0.01, 0.22+/-0.01 nM respectively. 4. The binding of [3H]-NMS was displaced dose dependently (pK50) by pirenzepine in CHO m1 membranes (7.97+/-0.04), methoctramine in CHO m2 membranes (8.55+/-0.1), 4 diphenylacetoxy-N-methyl piperidine methiodide (4-DAMP) in CHO m3 membranes (9.38+/-0.03), tropicamide in CHO m4 membranes (6.98+/-0.01). 4-DAMP, pirenzepine, tropicamide and methoctramine displaced [3H]NMS in CHO m5 membranes with pK50 values of 9.20+/-0.14, 6.59+/-0.04, 6.89+/-0.05 and 7.22+/-0.01 respectively. These data confirm homogenous subtype expression in CHO m1-m5 cells. 5. [3H]NMS binding was displaced dose-dependently (pK50) by pancuronium (m1, 6.43+/-0.12; m2, 7.68+/-0.02; m3, 6.53+/-0.06; m4, 6.56+/-0.03; m5, 5.79+/ 0.10), vecuronium (m1, 6.14+/-0.04; m2, 6.90+/-0.05; m3, 6.17+/-0.04; m4, 7.31+/ 0.02; m5, 6.20+/-0.07), pipecuronium (m1, 6.34+/-0.11; m2, 6.58+/-0.03; m3, 5.94+/-0.01; m4, 6.60+/-0.06; m5, 4.80+/-0.03), rocuronium (m1, 5.42+/-0.01; m2, 5.40+/-0.02; m3, 4.34+/-0.02; m4, 5.02+/-0.04; m5, 5.10+/-0.03) and gallamine (m1, 6.83+/-0.05; m2, 7.67+/-0.04; m3, 6.06+/-0.06; m4, 6.20+/-0.03; m5, 5.34+/ 0.03). 6. Cyclic AMP formation was inhibited dose dependently by methacholine in CHO m2 cells pEC50 for control and pancuronium (300 nM) treated cells were 6.18+/ 0.34 and 3.57+/-0.36 respectively. Methacholine dose-response curves in the absence and presence of rocuronium (1 microM) and vecuronium (1 microM) did not differ significantly. Pancuronium, vecuronium and rocuronium did not inhibit cyclic AMP formation alone indicating no agonist activity. 7. With the exception of rocuronium there was a significant interaction with m2 muscarinic receptors with all NMBD's at clinically achievable concentrations suggesting that the brady/tachycardias associated with these agents may result from an interaction with cardiac muscarinic receptors. Furthermore pancuronium at clinically achievable concentrations antagonised methacholine inhibition of cyclic AMP formation in CHO m2 cells further suggesting that the tachycardia produced by this agent results from muscarinic antagonism. The mechanism of the bradycardia produced by vecuronium is unclear. PMID- 9846650 TI - Characterization of protein serine/threonine phosphatase activities in human lung mast cells and basophils. AB - 1. The serine/threonine protein phosphatase (PP) inhibitors, okadaic acid and calyculin, attenuated the IgE-mediated release of histamine from human lung mast cells (HLMC) and basophils in a dose-dependent manner whereas an alternative PP inhibitor, microcystin, was ineffective. Calyculin was more potent than okadaic acid in both cell types. The concentration required to inhibit by 50% (IC50) the release of histamine was 15 (HLMC) and 50 nM (basophils) for calyculin and 200 (HLMC) and 300 nM (basophils) for okadaic acid. 2. Lysates of purified HLMC and basophils dephosphorylated radiolabelled glycogen phosphorylase, a substrate for both PP1 and PP2A. The PP activity in lysates of both cell types was inhibited in a dose-dependent fashion by the PP inhibitors with the following rank order of activity, calyculin (approximate IC50; 0.02-0.1 nM) > or = microcystin (0.1 nM) > okadaic acid (70 nM). 3. The PP1-selective inhibitor, inhibitor-2 (I-2), attenuated the dephosphorylation of glycogen phosphorylase in lysates of both HLMC and basophils. I-2 (20 nM) inhibited the glycogen phosphorylase PP activity by 71+/-3% and 49+/-13% in HLMC and basophil extracts, respectively. There were, approximately, 6 fold greater levels of I-2-sensitive activity in HLMC than in basophils. Qualitatively similar results were obtained with an alternative PP1 selective inhibitor, inhibitor-1 (I-1). 4. Lysates derived from HLMC and basophils dephosphorylated radiolabelled casein which is a PP2A-restricted substrate. HLMC lysates contained, approximately, 2.5 fold higher levels of casein PP activity than basophil lysates. 5. These data indicate that HLMC and basophils both contain PP1 and PP2A. The data suggest that, on a per cell basis, HLMC have higher levels of both PP1 and PP2A. Moreover, the ratio of PP1 to PP2A is higher in HLMC than in basophils. PMID- 9846651 TI - Expression of cyclo-oxygenase-2 in human airway smooth muscle is associated with profound reductions in cell growth. AB - 1. It is now accepted that uncontrolled proliferation of human airway smooth muscle (HASM) cells contributes, in many cases, to the chronic stages of asthma. However, the physiological and pathophysiological processes regulating cell growth and division in the airway are not clear. We have recently shown that the immediate early gene, cyclo-oxygenase-2, is induced by cytokines in HASM cells. Since cyclo-oxygenase metabolites, such as prostaglandin (PG) E2 have been shown to modulate HASM cell growth, we have investigated any autocrine action of endogenously released cyclo-oxygenase-1/2 products on the proliferative responses in these cells. 2. HASM cells were cultured from healthy tissue obtained at lung or heart/lung transplantation. HASM cell proliferation was measured by [3H] methyl thymidine uptake by cells and by cell counts. Cyclo-oxygenase-2 expression was measured by Western blot analysis and activity measured by the release of PGE2, by radioimmunoasay. 3. HASM cells proliferated in response to foetal calf serum, a response that was greatly inhibited when cyclo-oxygenase-2 was induced with either interleukin-1beta plus tumour necrosis factor-alpha or interleukin 1beta, tumour necrosis factor alpha plus interferon gamma (each at 10 ng ml(-1)). The inhibitory effect of cytokines on HASM cell proliferation was reversed in a concentration dependent manner by either the mixed cyclo-oxygenase-1/-2 inhibitor, indomethacin or the selective cyclo-oxygenase-2 inhibitor, L-745,337 (each at 10 microM). 4. PGE2 or the stable analogue of prostacyclin, cicaprost concentration-dependently (0.1 pmol to 1 microM) inhibited serum induced proliferation of HASM cells. By contrast, the TP receptor agonist, U46619 stimulated proliferation of HASM cells when cells were cultured without but not with serum. Other cyclo-oxygenase products, PGD2, PGF2alpha had no effect on cellular proliferation at concentrations up to 1 microM. 5. These observations illustrate a profound inhibitory effect of cyclo-oxygenase-2 induction on HASM cell proliferation, possibly via IP or EP receptor activation. Cyclo-oxygenase-2 induction has, thus far, been associated with the pro-inflammatory responses of plasma exudation and oedema formation and is assumed to be an enzyme worthy of selective inhibition in many disease states. However, our observations suggest that cyclo-oxygenase-2 can have an anti-inflammatory, anti-proliferative function in the airways. These observations may have importance in the use and development of therapies for airway disease such as asthma. PMID- 9846652 TI - Kinetics of leukocyte-induced changes in endothelial barrier function. AB - 1. Extravasation of polymorphonuclear leukocytes (PMN) and associated plasma leakage are key events in the inflammatory process. The kinetics of PMN-induced changes in endothelial barrier function were studied by means of confluent monolayers of bovine aorta or human umbilical vein endothelial cells (EC), cultured on permeable membranes and mounted in a two-compartment diffusion chamber. The model permitted continuous measurement of transendothelial electrical resistance (TEER), and analysis of protein efflux and PMN migration across the EC monolayer. 2. Transendothelial chemotactic stimulation (fMLP or LTB4) of PMN resting on EC in the upper compartment induced a prompt decline in TEER, followed by an increase in protein flux and transmigration of PMN. Adding the chemoattractant together with PMN in the upper compartment provoked adhesion of PMN, fall in TEER and increase in protein permeability, but no transmigration of PMN, whereas inhibition of PMN adhesion to EC by pretreatment with anti-CD18 mAb prevented all responses to chemotactic stimulation. 3. Chemoattractant induced adhesion of PMN to the EC monolayer induced a rapid rise in EC cytosolic free Ca2+, similar to that obtained by direct stimulation of EC with histamine, indicating an active response of EC to PMN activation and adhesion. 4. In summary, continuous recording of transendothelial electrical resistance in the in vitro model described permits rapid and sensitive analysis of leukocyte activation-induced effects on EC barrier function. The kinetics and specificity of the EC and PMN responses to chemoattractant stimulation suggest that activated PMN, via adhesion-dependent events, have a direct effect on EC junctional integrity independent of whether transmigration occurs or not. PMID- 9846653 TI - On the future of the speciality of Obstetrics and Gynecology. A Dutch view. PMID- 9846654 TI - Abnormal Doppler velocities in the umbilical artery. PMID- 9846655 TI - Etiology and prevention of pulmonary complications following beta-mimetic mediated tocolysis. AB - OBJECTIVES: This study documents biological (haematocrit variations) and therapeutic parameters (salbutamol doses, volumes perfused) in two groups tocolysed with salbutamol, one with and the other without APO in order to define the risk factors linked to APO and to establish a standard protocol of management. STUDY DESIGN: This retrospective study includes data from 68 intravenous salbutamol tocolysis with four resulting APOs, carried out between January 1st, 1993 and December 31st, 1995. RESULTS: There was an excessive level of salbutamol administered over 48 h in the complicated APO-group (122.5+/-52 mg) opposed to the non-APO group (44.9 21 mg) as well as an overload of perfused solute (3.1+/-1.11) versus (1.9+/-1.11). Blood hemodilution was demonstrated in the APO group with a decrease of haematocrit by over 10% between the admission and the control value. No other risk factor was found. CONCLUSION: Tocolysis should be administered at the lowest possible perfusion rate with incremental doses as long as the heart rate stays under 120 beats/min and stopped after 48 h. Administration of maximal 11 of solute perfused/day is recommended. For the patient's follow-up we estimate daily input and output fluid to avoid hydric overload, and a daily control of haematocrit whose variation must be less than 10%. PMID- 9846656 TI - Is conservative treatment of HELLP syndrome safe? AB - HELLP syndrome is associated with a high rate of maternal and perinatal morbidity and mortality, and often leads to immediate fetal extraction. However, this condition may occur very early in pregnancy and conservative approaches have been recently proposed. The limits of this approach are discussed with two cases of conservative management of HELLP syndrome complicated by abruptio placentae. PMID- 9846657 TI - Triptorelin acetate administration in early pregnancy: case reports and review of the literature. AB - When using a long protocol with cycle day 23 gonadotrophin-releasing hormone agonists (GnRH-a) administration, an elevated estradiol level or a missed period 10-14 days after initiating pituitary downregulation should alert the physician to the possibility of a pregnancy. We report 4 pregnancies occurring during pituitary downregulation with Triptorelin acetate in 366 in-vitro fertilization (IVF) cycles resulting in 3 deliveries of 4 normal neonates at term and 1 first trimester abortion. This supports published data reporting a 1% spontaneous pregnancy incidence in women undergoing pituitary desensitization GnRH-a during the luteal phase prior to planned IVF treatment, a 15.9% abortion rate and a 1.7% malformation rate. Our cases together with other published data suggest that pregnancy outcome is not adversely affected by GnRH-a administration during the luteal phase of the conception cycle. However, long term follow-up of these babies is still lacking and the number of reported cases is too small adequately to rule out the possibility of any detrimental effect related GnRH-a administration in pregnancy. PMID- 9846658 TI - Estonian medical birth registry 1992-1994: association of risk factors with perinatal mortality. AB - OBJECTIVE: To investigate the association of different maternal sociodemographic characteristics and infant sex with perinatal mortality among primiparas and multiparas. STUDY DESIGN: Analysis of routine data from the Estonian Medical Birth Registry covering the whole of Estonia. A total of 47 358 infants (including stillborns) with a birth weight 1000 g or more from 1992 through 1994 were studied. Perinatal mortality rate, crude odds ratio (OR) and adjusted OR (calculated by a logistic regression model) were used to evaluate the association. OR values were adjusted for maternal age at delivery, maternal ethnicity, educational level, residence, marital status, smoking status, history of previous abortion and infant sex. RESULTS: The perinatal mortality rate was 12.2 per 1000 total births among primiparous and 14.3 among multiparous women. The highest adjusted ORs of perinatal deaths were found in older (35 years and over) primiparas (1.78; 95% confidence interval (CI 0.88-3.57)) and multiparas (1.81; 95% CI 1.29-2.55), in unmarried (single) primiparas (1.59; 95% CI 1.14 2.20) and multiparas (1.98; 95% CI 1.29-3.05), in smoking primiparas (1.69; 95% CI 1.09-2.63) and multiparas (1.51; 95% CI 1.02-2.25), and in multiparas with unknown smoking status (1.98; 95% CI 1.18-3.33). CONCLUSION: The study provides further evidence that perinatal mortality is positively associated with increased maternal age, unmarried (single) status and smoking. PMID- 9846659 TI - Placental ratio in pregnancies at different risk for intrauterine growth. AB - OBJECTIVE: We have analyzed the placental/birthweight ratio in women at increased risk of intrauterine growth retardation and pregnancy-induced hypertension and in women with pregnancy 'complicated' by these conditions. STUDY DESIGN: A total of 89 women with small gestational age (SGA) infants, 355 with appropriate gestational age infants (200 in the uncomplicated pregnancy group) and 28 with large for gestational age (LGA) infants were considered. RESULTS AND CONCLUSION: The mean placental weight showed a significant increase from the SGA to the LGA in the two groups. The placental ratio tended to increase from the LGA group to the SGA one both in infants of women with uncomplicated pregnancy and with pregnancy complicated by intrauterine growth retardation or pregnancy-induced hypertension; these findings were statistically significant. PMID- 9846660 TI - Delayed interval delivery and survival of the two fetuses after second trimester loss of one triplet. AB - Due to the implementation of assisted reproduction techniques, the incidence of multiple pregnancies associated with fetal and neonatal complications has significantly increased. A woman in the 24th week of a triplet pregnancy came to the hospital because of premature rupture of membranes of one amniotic sack and she had a miscarriage of one of the fetuses the same day. After confirmation of the viability of the two fetuses, she was kept under observation with antibiotic therapy only. The woman gave live birth to these remaining fetuses in her 32nd week of pregnancy. The outcome of this case demonstrates that watchful expectancy may be a feasible alternative to invasive intervention. The aim of this report is to add to the currently very limited literature of an expectant (conservative) policy with regard to pregnancy outcome after the early loss of a fetus from a multiple pregnancy. PMID- 9846661 TI - Fetal cardiotocography and acid-base status during cesarean section. AB - OBJECTIVE: To assess the fetal well-being during cesarean section, in relation to the previous fetal condition. DESIGN: Observational study. SETTING: University Hospital. PATIENTS PARTICIPANTS: 204 women undergoing cesarean section (203 intrapartum cesarean sections): 177 with general anesthesia and 27 with spinal anesthesia. MATERIAL AND METHODS: We compared the fetal heart records of the last 30 min before cesarean section (during the first stage of labor) with those during cesarean section. Scalp blood analysis 30 min before the beginning of anesthesia induction was compared with umbilical artery analysis at delivery. MAIN OUTCOME MEASURES: Cardiotocography. Acid-base analysis. Apgar scores. RESULTS: During cesarean section there was a reduction in uterine activity, an increase in silent tracings and a decrease in late decelerations. Umbilical artery pH was lower than scalp pH (7.23+/-0.06 vs 7.30+/-0.06). Oxygen saturation was also lower (14.43+/-8.58% vs 18.99+/-8.4%). The values of pCO2 and of base deficit were higher. During cesarean section low values of modified Fischer scores were associated with low pH values of umbilical artery and low Apgar scores. CONCLUSION: Silent tracings appearing during cesarean section usually do not indicate fetal distress. Poor intracesarean fetal heart tracings were associated with worse indicators of neonatal well-being. Although umbilical pH were lower than scalp values, when the correction described in the literature was applied, the difference was of little clinical relevance. It is concluded that anesthesic, pharmacological and surgical events have slight repercussion in fetal well being. However, in a few cases fetal heart monitoring during cesarean section could detect otherwise undiagnosed cases of transient acidemia or depression in the fetus. PMID- 9846662 TI - Meconium stained amniotic fluid in very low risk pregnancies at term gestation. AB - OBJECTIVE: To determine the prevalence and clinical significance of meconium stained amniotic fluid (MSAF) in a low risk population at term gestation and to investigate whether MSAF is a predictor for intrapartum and neonatal morbidity. METHODS: A very low risk population including 37 085 consecutive deliveries at term composed the study population. A cross-sectional study was conducted and two groups of patients were identified according to the presence (n=6164) or absence (n=30921) of meconium in the amniotic fluid at delivery and the outcomes of the two groups compared. RESULTS: The prevalence of MSAF was 16.6%. The incidence of cesarean section (5.6% vs 2.3% P<0.01), instrumental deliveries (3.2% vs 1.8% P<0.01), fetal distress (6.5% vs. 2.1% P<0.01), clinical chorioamnionitis (0.2% vs. 0.1% P<0.01), post-partum infection (0.5% vs. 0.2% P<0.01), 1-minute Apgar score <3 (1.9% vs. 1.1% P<0.01), small for gestational age (7.4% vs. 6.4% P<0.01). was significantly higher in the MSAF compared with the clear amniotic fluid group. Intrapartum and neonatal mortality in this low risk population was significantly higher in the MSAF group (1.7/1000) compared with women with clear AF (0.3/1000). CONCLUSIONS: MSAF in a low risk population at term gestation is a predictor for adverse perinatal outcome and peripartum complications. PMID- 9846663 TI - Single intrauterine fetal death (fetus papyraceus) due to uterine trauma in a twin pregnancy. AB - Fetus papyraceus is a rare condition. We describe the intrauterine fetal death with subsequent fetus papyraceus of one twin due to blunt maternal trauma in the second trimester of pregnancy. Blunt maternal trauma should raise the clinical suspicion of fetal death and dictate further observation; in a twin pregnancy, the possibility of a single fetal death should be investigated. PMID- 9846664 TI - Declining perinatal mortality in Finland between 1987 and 1994: contribution of different subgroups. AB - OBJECTIVE: To examine the extent to which the decline in perinatal mortality is attributable to some subgroups, especially to certain birthweight or gestation groups. STUDY DESIGN: A register study using the Finnish Medical Birth Register for years 1987 to 1994. RESULTS: Of the overall reduction in perinatal mortality from 8.8 to 6.7 per 1000 births, 78% was due to stillbirths, compared with 22% due to early neonatal deaths. The decline in mortality among infants who weighed under 1500 g at birth was the major contributor (62%) to the overall reduction in perinatal mortality. The largest decline in mortality in the stillbirth group occurred among those weighing < 1000 g, while for early neonatal deaths the group most affected weighed 1000-1499 g. A similar pattern emerged when the gestation week groups were examined. CONCLUSION: The decline in perinatal mortality is attributable to stillbirths of very low birthweight. The most likely explanations for this result are the improved antenatal and neonatal care and the wider use of malformation screening. PMID- 9846665 TI - Risk factors associated with preterm (<37+0 weeks) and early preterm birth (<32+0 weeks): univariate and multivariate analysis of 106 345 singleton births from the 1994 statewide perinatal survey of Bavaria. AB - OBJECTIVE: The study was conducted to identify medical, obstetrical and social risk factors associated with early preterm births (<32+0 gestational weeks). STUDY DESIGN: The Statewide Perinatal Survey of Bavaria is a collection of perinatal data from all Bavarian maternity units using a uniform numbered questionnaire. Data on 106345 singleton births from the 1994 Survey were analysed using univariate and multivariate logistic regression analysis. RESULTS: In the multivariate analysis, early preterm birth was associated with premature rupture of the membranes (odds ratio (OR) 1.6, 95% confidence interval (CI) 1.37-1.86), treatment for infertility (OR 1.7, 95% CI 1.19-2.34), previous induced abortion (OR 1.8, 95% CI 1.57-2.13), maternal age >35 years (OR 1.8, 95% CI 1.47-2.16), premature cervical dilatation (OR 2.3, 95% CI 1.86-2.94), a history of stillbirth (OR 3.2, 95% CI 2.13-4.83), a history of preterm birth (OR 3.3, 95% CI 2.45 4.48), maternal age <18 years (OR 3.4, 95% CI 2.03-5.61), malpresentation (OR 3.9, 95% CI 3.10-4.93), preeclampsia (OR 4.0, 95% CI 3.20-4.94), uterine bleeding (OR 5.0, 95% CI 4.08-6.02), preterm labour (OR 7.0, 95% CI 5.94-8.22), and chorioamnionitis (OR 22.3, 95% CI 17.40-28.66). CONCLUSION: These data identify a subgroup of women at an increased risk for early preterm birth and may benefit from an intensified prenatal care. Risk factors related to the obstetrical history, genital infections, preeclampsia and maternal age are the most relevant for early preterm birth. PMID- 9846666 TI - Extremely rare type of conjoined twins: cephalothoracopagus deradelphus. AB - Occurrence of conjoined twins (CT) with a chromosomal abnormality is a very rare event. The case presented is that of a cephalothoracopagus Deradelphus twin with one cerebrum and two brain stems joined at the cranial end of the midbrain. This rare condition and its diagnosis and management are discussed with regard to counselling and obstetrical care. PMID- 9846667 TI - Extracorporeal membrane oxygenation--a last-resort therapy for newborns with congenital diaphragmatic hernia or other causes of severe respiratory failure. PMID- 9846668 TI - Pure lipoma of the uterus in association with endometrial carcinoma. AB - Pure lipoma of the uterus is a very rare entity, with few cases described in the English literature. We report the case of a 71-year-old woman, with pure lipoma of the uterus and coexistent endometrial carcinoma and discuss the possible relationship between these pathologic entities. PMID- 9846669 TI - Vesico-vaginal fistula in Jordan. AB - In order to study the aetiology of vesico-vaginal fistula in Jordan, and assess the outcome of surgical treatment, 132 cases of acquired vesico-vaginal fistulae treated at Al-Bashir Hospital, and Jordan University Hospital, Amman, Jordan, during the period between 1972 and 1996 were reviewed. Forty-five cases (30%) had one or more previous failed repair outside these hospitals. Successful repair of 108 cases was achieved (82%), 60 cases of them from the first attempt (45%) and, 48 other cases after repeated attempts (36%). In 24 cases, a total of 72 repeated operations failed. Obstetric fistulae in Jordan are decreasing due to improvement and expansion of maternity care. Although a patient with neglected fistula is a social problem, this is not the case in Jordan because of the strong family ties. Nevertheless, they present a great challenge to the gynaecologist. Referral to a centre experienced in this type of surgery certainly improves the success rate, and diminish the suffering of these patients. PMID- 9846670 TI - Pre-operative ultrasonographic evaluation of nongravid, enlarged uteri- correlation with bimanual examination. AB - OBJECTIVE: To correlate the size of the nongravid uterus measured by ultrasound and bimanual examination before hysterectomy, with the actual size of the specimens. PATIENTS AND METHODS: Uterine size of 49 consecutive patients, who underwent elective hysterectomy, was assessed by bimanual pelvic examination, preoperative ultrasonographic evaluation and actual postoperative size. RESULTS: All preoperative ultrasonographic uterine dimensions significantly correlated with the corresponding actual dimensions of the uterine specimens. Furthermore, ultrasonographic uterine length had the best correlation with uterine size estimation by bimanual examination. A formula was established in an attempt to calculate clinical uterine size by gestational week, using preoperative ultrasonographic dimensions. CONCLUSIONS: By using the aforementioned formula, ultrasonographic uterine dimensions can be extrapolated to uterine size in terms of gestational week, and therefore, be applicable to traditional clinical practice. PMID- 9846671 TI - Prognostic factors of the uterine cervix adenocarcinoma. AB - BACKGROUND: Prognosis factors for adenocarcinoma of the uterine cervix after primary treatment are poorly established. METHODS: A retrospective study of 45 cases of adenocarcinoma of the cervix with a follow-up of 96 months on average was performed. The primary treatment consisted in combined radical surgery and radiotherapy for stage I-II patients while patients with advanced disease were treated by radiotherapy. In case of poor prognosis factors, they were given chemotherapy. Survival rates were established and prognosis factors influencing survival and recurrences were studied. RESULTS: Fifteen women remained alive without evolutive disease. FIGO stage and pelvic node involvement were the most important parameters influencing overall survival. Local failures (27%, average period of 30 months) were unpredictable and led to a dramatic outcome. Histological grade and pelvic node status were significant predictive factors for metastatic recurrence (40%, average period of 29 months). CONCLUSIONS: Local recurrence and metastatic dissemination of cervical adenocarcinoma after primary treatment prove to be rapidly fatal although life expectancy can be prolonged with adjuvant treatment of the recurrence. In the event of aggressive tumors with high histological grade and pelvic node involvement, an attempt to assess adjuvant systemic chemotherapy could be useful. PMID- 9846672 TI - Pudendal canal syndrome as a cause of vulvodynia and its treatment by pudendal nerve decompression. AB - Notwithstanding many established causes of vulvodynia there still remains an idiopathic group with unknown etiology and variable results of treatment. We present 11 women with idiopathic vulvodynia in whom the etiology could be defined and who were successfully treated. Age varied from 28-53 years. The vulvar pain was associated with stress urinary incontinence in 6/11 patients and all had constipation. Perineal and vulvar hypoesthesia occurred in 6, weak anal reflex in 7 and diminished EMG activity of the external anal sphincter in 3, of the external urethral sphincter in 6 and of the levator ani muscle in 11. There was significant increase (P<0.05) of the pudendal nerve terminal motor latency (PNTML) in all. The motor and sensory change as well as the increased PNTML point to pudendal canal syndrome. Pudendal nerve block, as a diagnostic and therapeutic test, effected temporary pain relief. Pudendal nerve decompression was performed. The inferior rectal nerve was exposed through a para-anal incision, and followed to the pudendal nerve in the pudendal canal. Pudendal canal fasciotomy was done to release the pudendal nerve in the ischiorectal fossa. Vulvar pain disappeared in 9/11 women and stress urinary incontinence in 4/6. Anal reflex was normalized in 5/7 women, and vulvar and perineal hypoesthesia in 4/6. The EMG activity of the external urethral sphincter improved in 4/6, of the external anal sphincter in 2/3 and of the levator ani in 9/11 women. The PNTML was normalized in 9/11 women. In conclusion, pudendal nerve decompression effected relief and improvement in the sensory and motor manifestations of the pudendal nerve in 9/11 women. Two women did not improve due probably to an irreversible damage of the pudendal nerve, or to incomplete pudendal nerve decompression. PMID- 9846673 TI - Tamoxifen and proliferation of vaginal and cervical epithelium in postmenopausal women with breast cancer. AB - OBJECTIVE: The estrogenic effect of tamoxifen on vaginal and cervical epithelium in postmenopausal women with breast cancer is evaluated. STUDY DESIGN: The tamoxifen group consisted of 92 postmenopausal breast cancer patients, the control group I of 30 postmenopausal women with breast cancer receiving no endocrine therapy and the control group II of 40 postmenopausal women without primary breast cancer taking no hormones. We determined the maturation index and the incidence of endocervical cell hyperplasia and metaplasia in cervical and vaginal smears. RESULTS: The maturation index increased under tamoxifen within the first 24 months from 0.4011 before taking tamoxifen (n=56) to 0.6039 (n=138, P<0.0001). The maturation index in the group treated with tamoxifen was statistically significantly higher (P<0.0001) than in the control groups (control group I: 0.3975, P<0.0001; control group II: 0.4102, P<0.0001). Under therapy with tamoxifen endocervical cell hyperplasia (P=0.00156) and metaplasia (P=0.00123) appeared significantly more often. CONCLUSION: An apparent increase not only of the incidence of endocervical cell hyperplasia and metaplasia but also of the maturation of the vaginal epithelium caused by the estrogenic effect of tamoxifen could be demonstrated. PMID- 9846674 TI - Genital tuberculosis in a HIV infected woman: a case report. AB - The incidence of HIV-associated tuberculosis is increasing worldwide, especially in developing countries. HIV infected patients rapidly develop clinically significant disease, respond poorly to complete treatment and present with extrapulmonary tuberculosis. Although a relative increase of genital tuberculosis would be expected, this has not been reported. Probably, tuberculous systemic disease is diagnosed earlier, before genital tuberculosis occur. The present study is a report of case of a young African female patient, who was admitted with symptoms of acute pelvic inflammatory disease due to genital tuberculosis and proved to be HIV infected. The patient was managed by intravenous antibiotic administration, but since no clinical or laboratory improvement was achieved, a laparotomy and salpingooophorectomy was performed. Histopathology revealed tuberculosis and after that the patient proved to be HIV infected. Further investigation did not reveal pulmonary or other extragenital manifestation of tuberculosis. The only manifestation of HIV infection and genital tuberculosis was the symptoms of an acute pelvic inflammatory disease, which is extremely rare. PMID- 9846675 TI - The frequency of bacterial and yeast infection in women with different grades of cervical intraepithelial neoplasia (CIN). AB - OBJECTIVE: To determine bacterial and yeast infection of the uterine cervix in women with different grades of cervical intraepithelial neoplasia (CIN). STUDY DESIGN: 578 patients with CIN were included in this study. In order to determine the presence of bacterial and yeast infection, a cervical swab was obtained before conization of the uterine cervix. After surgery and the definitive histology report, the frequency of bacterial and yeast infection in different grades of CIN was calculated. RESULTS: Among 578 patients with CIN, bacterial or yeast infection was present in 379 (65.6%) patients. In patients with CIN 1, infection was present in 20 (71.4%), in CIN 2 in 106 (69.7%) and in CIN 3 in 252 (63.3%) cases. The differences in the frequency of infection among all three groups are not significant. CONCLUSION: In patients with CIN bacterial and yeast infection of the uterine cervix is very common. Its occurrence does not depend on the grade of CIN. PMID- 9846676 TI - Effects of tibolone on the breast. AB - OBJECTIVE: to evaluate the effect of hormone replacement therapy and tibolone on the breast. STUDY DESIGN: prospective, controlled, randomized study. SETTING: Outpatient Menopause Clinic of the Second University of Naples. PARTICIPANTS: forty four women in spontaneous menopause without any risk factor for breast cancer were randomly allocated to three groups: 15 patients (group A) were treated with transdermal oestrogens 50 microg, 2 patches/week for 3 weeks per month, plus acetate nomegestrolo per os 5 mg/die for 12 days per cycle, 17 patients (group B) were treated with tibolone 2.5 mg/die. Twelve patients not given any medication represented the control group (group C). METHODS: at the time of recruitment and after at least 12 months of therapy the patients were subjected to a questionnaire aimed at quantifying the slight, moderate or severe presence of the tension/mastodynia symptoms and to a mammographic test to assess the parenchymal pattern according to a quantitative method: type 1 (less than 25% of mammary gland covered by dense tissue), type 2 (from 25% to 75% of total glandular area covered by dense tissue), type 3 (more than 75% of mammary parenchyma covered by dense tissue). Statistical analysis was carried out by means of Fisher's exact test. RESULTS: after at least 12 months of treatment in Group A 5 out of 15 patients (33%) showed a trend of increase in mammographic density not statistically significant (P=0.22) when compared with group B in which one patient showed a swift from type 1 to type 2 and another from type 2 to type 3. The analysis of tension/mastodynia symptoms revealed a significantly difference between the two groups (P=0.02): in group A mastodynia appeared in three previously asymptomatic women and increased in five women, with a total increase in the symptomatology in 8 out of 15 patients (53.3%), in group B only in one case (5%) mastodynia turned from slight to moderate. CONCLUSION: in postmenopausal women oestroprogestogenic replacement therapy may be associated with an increase in mammographic density and with the onset or increase in mastodynia. On the contrary tibolone does not seem to affect normostructured mammas and may be considered a first-rate replacement therapy in case of mammas showing particular density or benign mastopathies. PMID- 9846677 TI - Violence against women: the physician's role. AB - Violence against women is one reflection of the unequal power relationship between men and women in societies. Reflections of this inequality include marriage at a very young age, lack of information or choice about fertility control and forced pregnancy within marriage. The different forms of violence against women are: domestic violence and rape, genital mutilation or, gender based violence by police and security forces, gender-based violence against women during armed conflict, gender-based violence against women refugees and asylum seekers, violence associated with prostitution and pornography, violence in the workplace, including sexual harassment. Violence against women is condemned, whether it occurs in a societal setting or a domestic setting. It is not a private or family matter. The FIGO Committee for the Study of Ethical Aspects of Human Reproduction released statements to physicians treating women on this issue. Physicians are ethically obliged to inform themselves about the manifestations of violence and recognize cases, to treat the physical and psychological results of violence, to affirm to their patients that violent acts toward them are not acceptable and to advocate for social infrastructures to provide women the choice of seeking secure refuge and ongoing counselling. PMID- 9846678 TI - The sperm deformity and the sperm multiple anomalies indexes in patients who underwent unilateral orchectomy and preventive radiotherapy. AB - Two new indices, the sperm deformity and the sperm multiple anomalies index, were assessed in 16 male patients who underwent unilateral orchectomy owing to seminoma and in preventive radiotherapy of paraortic and of the iliac lymph glands homolateral to the testis excised. The aim was to assess the value of these two morphological sperm parameters in distinguishing the semen between the above mentioned group of men from the semen of men with other causes of infertility. It was found that: (1) the values of the sperm deformity index and those of the sperm multiple anomalies index were not significantly different between men who underwent unilateral orchectomy and preventive radiotherapy and the men with other causes of subfertility and (2) the values of the above indices were significantly higher in the semen specimens of men who underwent unilateral orchectomy and preventive radiotherapy and in the semen specimens of men with other causes of subfertility than in the semen specimens of fertile men. It is concluded that testicular response, as far as sperm morphology is concerned, is the same irrespective of the cause exerting a negative effect on spermatogenesis. PMID- 9846679 TI - The effect of the duration of GnRH-agonist down regulation before ovarian stimulation on the biological and clinical outcome after intracytoplasmic sperm injection. AB - The object of this study was to compare the biological outcome (oocyte maturity, fertilization, cleavage) and the clinical outcome after a 'long' (15-24 days) and a 'long-long' (25-40 days) protocol of GnRH-agonist administration for intracytoplasmic sperm injection. Group A consisted of 51 patients with a 15-24 day down regulation period and Group B consisted of 35 patients with a 25-40-day down regulation period, all of which entered ICSI due to severe male factor infertility. Duration and amount of gonadotropin stimulation, serum E2 on the day of hCG administration, number of oocytes retrieved, oocyte maturity, fertilization rate, cleavage rate and pregnancy outcome were comparable for the two groups of patients. Therefore, a flexible period of pituitary desensitization can be employed, allowing us to simplify planning for patients and for the medical staff without affecting the outcome of the trial. PMID- 9846680 TI - Timed intercourse after intrauterine insemination for treatment of infertility. AB - OBJECTIVE: To compare the pregnancy rates, between intrauterine insemination (IUI) followed by timed intercourse and IUI only for treatment of the infertile couples. STUDY DESIGN: A prospective study of two different protocols of intrauterine insemination in two hundred and one infertile couples with a normal spermiogram was carried out. Of these, 101 couples were treated with IUI alone and 100 couples had both IUI and timed intercourse within a 12-18 h period. The pregnancy rates were compared between groups. RESULTS: The characteristics of the two groups were similar in terms of the mean age, as well as the duration and causes of infertility. The cycle characteristics following follicular stimulation were also similar between two groups. The pregnancy rate per cycle increased with increasing numbers of total motile sperm per insemination in the IUI alone group (P=0.045). Timed intercourse increased pregnancy rate in patients with lower motile sperm number (<40x10(6)) (27.7% versus 10.5%, P=0.023), but not in patients with higher sperm number (> or =40x10(6)) (25.7% versus 22.7%, P=0.671). CONCLUSIONS: In IUI with low number of motile sperm inseminated, timed intercourse significantly increases the pregnancy rates over IUI alone in infertile couples with a normal sperminogram. This alternative treatment appears to be a practical, simple, and inexpensive addition that improves the pregnancy rate in patients receiving ovulation induction and intrauterine insemination program. PMID- 9846681 TI - Placental teratoma. A case report. AB - We present a rare case of a placental teratoma found within the fetal membranes of an otherwise normal placenta. Lack of a recognizable umbilical cord and no recognizable skeletal development are used as differential diagnostic criteria for a placental teratoma. In our case, an umbilical cord was absent; nevertheless, the tumor contained two vascular channels covered only by fetal membranes without Wharton's jelly. PMID- 9846682 TI - Laparoscopic surgery for heterotopic pregnancies: a case report and a brief review. AB - A heterotopic pregnancy is in effect a multiple pregnancy with one or more intrauterine pregnancies coexisting with an ectopic pregnancy and is rarely spontaneous. With the increasing popularity of ovulation induction performed during assisted reproductive techniques, it will not be surprising to observe that this phenomenon has increased significantly. However, diagnosis is often delayed because of its rarity and difficulty. We report a case of a woman with a viable intrauterine pregnancy who had a complication of ovarian hyper-stimulation syndrome secondary to ovulation induction following in vitro fertilization and embryo transfer, but who, during hospitalization, presented with clinically progressive abdominal pain. An unruptured ectopic pregnancy of the right fallopian tube was diagnosed accidentally by laparoscopy and laparoscopic salpingectomy was immediately performed. Post-operative follow-up revealed that the intrauterine pregnancy continued normally. She delivered a normal female baby at 38 weeks of gestation. The promising neonatal outcome might suggest that laparoscopy might be safely performed to aid differential diagnosis in an uncertain condition during pregnancy: therefore, laparoscopic surgery might be an appropriate method to manage some carefully selected patients with HP. A brief review of the published literature on the role of laparoscopy in the diagnosis and management of heterotopic pregnancy is given. PMID- 9846683 TI - Hyperemesis in late pregnancy--should we think of cancer? A case report. AB - Gastric cancer is unusual during pregnancy. The diagnosis may be delayed because specific symptoms are similar to typical pregnancy associated complaints. Our therapeutic management with palliative chemotherapy and later gastrectomy differs from other known cases, where surgical resection has been the treatment of choice. Surgery appears to have no influence on the prognosis of gastric cancer patients with hepatic metastases. PMID- 9846684 TI - Double maternal seroconversion to cytomegalovirus and Toxoplasma gondii. AB - BACKGROUND: We report the first case of a double maternal seroconversion for Toxoplasma gondii (TG) and cytomegalovirus (CMV) diagnosed during pregnancy. CASE: One case is reported of a female patient referred for seroconversion in response to TG in the 27th week of gestation. A search for foetal involvement revealed signs of non-specific foetal infection without any TG-related lesions. Tests were carried out for another foeto-maternal infectious disease and maternal seroconversion in response to CMV was discovered with virus in the amniotic fluid. The foetus developed hydrocephalus and intracranial calcifications and the pregnancy was terminated at the parents' request. CMV-induced multiple organ involvement without any signs of Toxoplasma gondii-related involvement were noted in the foetus. CONCLUSIONS: This case indicates that a search should be made for another infectious disease likely to involve the foetus when non-specific signs of infection in the foetus are present, even though maternal seroconversion has been recognized. PMID- 9846685 TI - Gunshot uterine rupture: a case report. AB - A case report of gunshot uterine rupture from which the mother and fetus survived is presented for the first time. Management was guided by pre-operative ultrasound diagnosis of traumatic anhydramnios. PMID- 9846686 TI - Hereditary thrombophilia in a family with three independent protein S and C mutations. A cause of adverse perinatal outcome. AB - A non-related couple with two independent protein S and a protein C mutation had two of their three children suffering from severe thrombosis resulting in neonatal death of the firstborn. With prenatal testing an accurate prediction of phenotype was possible for the second child but the third infant was more severely affected than had been predicted from the genotype. PMID- 9846687 TI - Transient fetal cerebral dysfunction after road traffic accident. A case report. AB - We present a case at 30 weeks gestation with transient abnormal fetal cerebral function after a road traffic accident, with loss of fetal movements and abnormal FHR tracings for almost 1 week. The pregnancy had after this incident a normal course and a healthy baby boy was born at term. We suggest the mechanism to be related to fetal hypoxia after excessive maternal psychological stress. PMID- 9846688 TI - High-level IL-12 production by human dendritic cells requires two signals. AB - IL-12 is a key cytokine in the development of Th1 responses. IL-12 production by antigen-presenting cells (APC) can be induced by the interaction between CD40 on the APC and CD40 ligand (CD40L) expressed on T cells after activation. Our previous study indicated that in dendritic cells (DC), the only APC that can activate naive T(h) cells efficiently, the mere CD40 engagement is insufficient to induce IL-12 production. The aim of the present study was to dissect the conditions for efficient IL-12 production by DC further. Using populations of naive and memory Th cells, recombinant CD40L, neutralizing and blocking antibodies, and by determining IFN-gamma production and CD40L expression levels, we here show that T cell-induced IL-12 production by DC results from the action of two signals, mediated by CD40L and IFN-gamma, and that the inability of naive T(h) cells to induce IL-12 production resides in their inability to produce IFN (gamma). Other factors than CD40L and IFN-gamma can provide the required signals for IL-12 production by DC, as either factor could be replaced by lipopolysaccharide (LPS). The two-signal requirement proved unique for the production of IL-12, since either CD40 engagement or LPS was sufficient for the efficient production of tumor necrosis factor-alpha, IL-8 and the p40 subunit of IL-12, and may be considered as a safety mechanism for optimal control of potentially harmful T(h)1 responses. PMID- 9846689 TI - Rapid synthesis of IFN-gamma by T cells in skin may play a pivotal role in the human skin immune system. AB - This study has shown that T lymphocytes with alphabeta TCR present in normal human skin possess unique characteristics which may allow them to fulfil a unique role in skin. It disclosed that unlike T cells from blood, they are devoid of proliferative capacity in response to any ligand including lectins and phorbol esters, and show a diminished Ca2+ flux in response to phytohemagglutinin (PHA). However, despite this apparent lack of reactivity, these cells respond to stimulation by synthesis of IFN-gamma. Furthermore, this response by T cells from the skin differs from that of T cells in blood in its tempo. Whereas IFN-gamma is first detectable in peripheral blood lymphocytes (PBL) 42 h following stimulation, T cells from skin produce IFN-gamma 2.5 h after activation, or 17 times faster. Whereas synthesis persists in PBL for several days, it only lasts in skin T cells for 90 min. This difference in the kinetics is a consequence of differences in the regulation of IFN-gamma synthesis in PBL and skin-derived T cells. Thus, unlike PBL, T cells in intact unimmunized skin or ex vivo express mRNA for IFN-gamma, transcribed in a constitutive manner. Activation of these cells turns on translation of IFN-gamma synthesis directly and at the same time turns off further transcription of the IFN-gamma gene: cessation of IFN-gamma synthesis coincides with the disappearance of IFN-gamma mRNA within 4 h of activation. These observations suggest that this pre-formed mRNA may be solely responsible for the early synthesis of IFN-gamma in skin, determining both the amount and the duration of its synthesis. We propose that these cells may represent the antigen-specific component of a first-line host defence system in human skin. The reported absence of delayed-type hypersensitivity reactivity in patients with a selective deficit of skin T cells supports this conclusion. PMID- 9846690 TI - Expression of a type II collagen-specific TCR transgene accelerates the onset of arthritis in mice. AB - Animal models of autoimmune diseases have been instrumental in advancing our understanding of autoimmunity in humans. Collagen-induced arthritis in mice is an autoimmune disease model of rheumatoid arthritis, which is MHC class II restricted and CD4 T cell dependent. To better understand the fundamental role of T cells in arthritis, we have generated a transgenic mouse carrying the rearranged Valpha11.1 and Vbeta8.2 TCR chain genes isolated from a type II collagen (CII)-specific T cell hybridoma. Cell surface analysis indicated that Vbeta8.2 chain was expressed on the surface of nearly all peripheral T cells. Analysis of T cell subsets in transgenic mice revealed a profound skewing in peripheral T cells towards the CD4 population. Although peripheral T cells were not tolerant to CII and responded to CII stimulation in vitro, transgenic mice did not develop spontaneous arthritis. However, a rapid onset of arthritis with severe clinical signs was detected in transgenic mice after immunization with CII in complete Freund's adjuvant. Histological analysis of inflamed joints showed a great resemblance to arthritic joints in man. This unique transgenic mouse model provides valuable insights into the mechanism of arthritis and into potential specific immune interventions. PMID- 9846691 TI - Precocious expression of T cell functional response genes in vivo in primitive thymocytes before T lineage commitment. AB - The genes encoding effector molecules of mature T cells, IL-2, perforin and IL-4, were found to be expressed in vivo in the most primitive subsets of thymocytes of adult mice. These subsets have previously been identified by their cell surface markers and by their expression of other T lineage-associated genes. While IL-2, perforin and IL-4 are expressed in distinct patterns, all three are expressed before the induction of RAG-1 and pre-TCR alpha mRNA expression, and are confined to subsets of cells that apparently have not yet undergone commitment to the T lineage. Thus, expression of T cell response genes appears to be one of the earliest markers of lymphocyte differentiation. Activation events marked by CD69 induction occur in these early cell types, but the response gene expression by these cells is separable from CD69 expression. IL-2 and perforin are induced again much later in thymocyte development, during TCR-dependent repertoire selection. At those stages, IL-2 protein and RNA levels per cell are higher, but the fraction of cells expressing IL-2 appears to be much lower than in the most immature stages. In addition, a striking feature of the immature populations is the robust IL-2 expression by presumptive immature NK cells. These findings are discussed in terms of the developmental origins of lineage specificity in T cell response gene regulation. PMID- 9846692 TI - The induction of transplantation tolerance by intrathymic (i.t.) delivery of alloantigen: a critical relationship between i.t. deletion, thymic export of new T cells and the timing of transplantation. AB - Intrathymic (i.t.) injection of donor alloantigens has proved to be an effective strategy for the induction of tolerance. However, the mechanisms by which tolerance is induced and maintained after transplantation remain unclear. In this report we show that tolerance to donor cardiac allografts can be induced across a MHC class I difference by i.t. injection of donor splenocytes and transient T cell depletion. Furthermore, using H-2K(b)-specific TCR transgenic mice (BM3), we demonstrate that prolonged deletion of donor-reactive thymocytes was essential to induce tolerance by i.t. injection and this was dependent upon donor cells persisting in the thymus. Examination of the kinetics of thymic export following i.t. injection revealed that prolonged deletion of thymocytes was required to delay export of new T cells to the periphery until the time of transplantation. Importantly, after transplantation donor cell persistence in the thymus and i.t. deletion were no longer necessary to maintain tolerance. The graft itself or cells from the graft was responsible for maintaining tolerance at this stage. These findings reveal that multiple mechanisms are responsible for the induction and maintenance phases of tolerance to alloantigens in vivo after i.t. delivery, and that a complex inter-relationship between donor cell persistence in the thymus, i.t. deletion, thymic export of T cells and the timing of transplantation is involved. PMID- 9846693 TI - Both CD80 and CD86 co-stimulatory molecules regulate allergic pulmonary inflammation. AB - We examined the roles of CD80 (B7-1) and CD86 (B7-2) in a model of allergic pulmonary inflammation and airway hyper-responsiveness (AHR) by selectively inhibiting either CD80 or CD86. Inhibition of co-stimulation by either CD80 or CD86 affected multiple parameters of the allergic response. Specifically, blockade of either CD80 or CD86 in ovalbumin-sensitized and challenged mice resulted in reduced expression of IL-2Ralpha (CD25) on CD4+ T lymphocytes, decreased airway eosinophilia, lower serum IgE production and diminished AHR. Importantly, blockade of CD80 and CD86 inhibited production of IL-4 and IL-2, and enhanced IFN-gamma production. Our observations support a role for both CD80- and CD86-mediated co-stimulation in development of allergic pulmonary inflammation. PMID- 9846694 TI - Fibronectin-binding protein I of Streptococcus pyogenes promotes T cell independent proliferation of murine B lymphocytes and enhances the expression of MHC class II molecules on antigen-presenting cells. AB - We have previously shown that fibronectin-binding protein I (SfbI) of Streptococcus pyogenes can act as an adjuvant for mucosal-delivered antigens (Medina, E., Talay, S. R., Chhatwal, G. S. and Guzman, C. A. 1998. Eur. J. Immunol. 28:1069). To characterize the underlying mechanism of the adjuvancity, we investigated the in vitro stimulating activity of SfbI. The SfbI protein promoted a dose-dependent proliferation of mouse spleen cells. Studies performed using cellular subpopulations showed that proliferation involved B cells and was T cell- and macrophage-independent. SfbI also induced lg production by B cells in a T cell-independent manner. The kinetics of lg isotype accumulation in supernatant fluids and the analysis of Ig-secreting cells suggested that SfbI stimulates B cells expressing different Ig isotypes rather than promoting the isotype switching of single subpopulations. Experiments performed with recombinant proteins encompassing different functional domains of SfbI showed that the fibronectin-binding repeats were responsible for B cell activation. The sera from mice immunized by the intranasal route with SfbI did not react with either double-stranded DNA, cardiolipin or collagen. Interestingly, stimulation with Sfbl also resulted in the up-regulation of MHC class 11 molecules expression by B cells and macrophages. The elucidation of the underlying molecular events to the immunomodulatory effect exerted by SfbI will facilitate the exploitation of the potential of this molecule for the generation of mucosal vaccines. PMID- 9846695 TI - Cell lines transfected with the TAP inhibitor ICP47 allow testing peptide binding to a variety of HLA class I molecules. AB - The immediate early protein ICP47 of the Herpes simplex virus is known to block the human transporter associated with antigen processing (TAP), thereby creating a TAP-deficient phenotype in any human cell transfected with the corresponding cDNA. Exploiting this inhibitory activity, we constructed a selection of human cell lines each co-expressing one of the cDNAs of human leukocyte antigen (HLA) class I alleles HLA-A*1101, A24, A*3101, A*6601, B8 and B*1516, and the cDNA encoding the ICP47 molecule. The cell lines generated showed diminished HLA class I surface expression and the inhibition of the TAP function was confirmed in peptide translocation assays. The addition of specific exogenous peptide ligands restored the expression of the corresponding HLA class I molecules. Thus, the ICP47 transfectants provide us with a tool to closely examine peptide-HLA class I interactions, to confirm HLA class I ligand motifs and to test peptides predicted to bind. PMID- 9846696 TI - B cell receptor-induced apoptosis in primary transitional murine B cells: signaling requirements and modulation by T cell help. AB - Self-reactive immature B cells may be eliminated in the bone marrow (BM) after B cell receptor (BCR) engagement in a process known as negative selection. Immature B cells emigrating from the BM, the so-called transitional cells, remain sensitive to negative selection and are likely to be important targets of tolerance towards peripheral antigens. Transitional cells are deleted through apoptosis after BCR cross-linking in vitro. Using anti-Ig as a surrogate antigen, we determined the signaling requirements for the induction of apoptosis in transitional cells. Treatment with anti-Ig for only 20 min causes most cells to be apoptotic 16 h later. Furthermore, apoptosis of transitional cells is induced with low doses of anti-Ig while mature cell proliferation requires extended culture at 30-fold higher concentrations. For both populations of B cells, total surface Ig expression is equivalent, therefore indicating that the threshold of BCR signaling required to elicit these responses is different. T cell help can modulate B cell tolerance. However, specific help may not be available when apoptosis is triggered by a peripheral antigen. The opportunity to reverse apoptosis of transitional cells is surprisingly long. Even 8 h after anti-Ig treatment, IL-4 or anti-CD40 antibody can block apoptosis. The upper time limit of protection is concurrent with irreversibility of apoptosis as measured by DNA fragmentation. These findings indicate that B cell negative selection is more easily triggered than activation, and that the induction of apoptosis and its reversal by T cell help can be events that occur in distinct microenvironments. PMID- 9846697 TI - An expressed neo(r) cassette provides required functions of the 1gamma2b exon for class switching. AB - Germline CH transcripts initiate from a promoter upstream of a non-coding I exon, proceed through the switch (S) region and terminate downstream of the associated CH exons. To elucidate the role of germline transcription in Ig heavy chain class switch recombination (CSR), we used gene targeting in embryonic stem (ES) cells to replace most of the Igamma2b exon from immediately 3' of the majority of transcription initiation sites to beyond its donor splice site with a PGK neo(r)gene inserted in the same transcriptional orientation as the endogenous unit. The mutation was introduced into both alleles of ES cell lines (referred to as gamma2-b(N/N)) and the neo(r) gene was deleted (referred to as Igamma2b-/-) by the loxP/Cre method. These mutations were assayed for effects on CSR in B cells derived via RAG-2-deficient blastocyst complementation. Igamma2b-/- B cells lacked ability to switch to IgG2b both in vivo and in vitro, and, correspondingly, showed no germline transcription through the Igamma2b exon, Sgamma2b or the Cgamma2b region. In contrast, Igamma2b(N/N) B cells switched at normal levels to IgG2b and showed substantial transcription through the Sgamma2b and Cgamma2b regions. Taken together, these results show that the Igamma2b sequences, per se, are not necessary for mediating CSR since a transcribed PGK neo(r) gene can replace its function. However, the deleted portion of the Igamma2b exon and splice donor site apparently contain sequences necessary for efficient germline gene transcription and thus for CSR to IgG2b. PMID- 9846698 TI - The characterization of murine BCMA gene defines it as a new member of the tumor necrosis factor receptor superfamily. AB - The BCMA gene is a new gene discovered by the molecular analysis of a t(4;16) translocation, characteristic of a human T cell lymphoma. It has no significant similarity with any known protein or motif, so that its function was unknown. This report describes the cloning of murine BCMA cDNA and its genomic counterpart. The mouse gene is organized into three exons, like the human gene, and lies in murine chromosome 16, in the 16B3 band, the counterpart of the human chromosome 16p13 band, where the human gene lies. Murine BCMA cDNA encodes a 185 amino acids protein (184 residues for the human), has a potential central transmembrane segment like the human protein and is 62% identical to it. The murine BCMA mRNA is found mainly in lymphoid tissues, as is human BCMA mRNA. Alignment of the murine and human BCMA protein sequences revealed a conserved motif of six cysteines in the N-terminal part, which strongly suggests that the BCMA protein belongs to the tumor necrosis factor receptor (TNFR) superfamily. Human BCMA is the first member of the TNFR family to be implicated in a chromosomal translocation. PMID- 9846699 TI - Survival of long-lived plasma cells is independent of antigen. AB - Recent studies have shown that persistent specific antibody titer is provided by long-lived plasma cells (PC) which constitute a new kind of 'memory-providing cells'. In the present study, we examine the role of antigen for the long-term survival of PC and the maintenance of specific serum antibody titers. Using a novel cytometric technology, to identify and isolate antigen-specific PC, we analyzed long-lived PC of BALB/c mice, during their development (between day 1 and 10) after secondary immunization with ovalbumin (OVA) and in the phase of the established immune reaction. Most if not all OVA-specific PC were generated within a few days after immunization. Within approximately 3 weeks, they matured, as indicated by down-regulation of expression of MHC class II. These PC are long lived and located in spleen and bone marrow. Upon adoptive transfer, OVA-specific PC from bone marrow, but not memory B cells, conferred specific and long-lasting antibody titers to antigen-free IgH syngeneic recipients. In response to antigenic challenge, new OVA-specific antibody-secreting cells were generated from transferred memory B cells. Antibody secretion by long-lived PC was not affected. Our results confirm that persistent antibody titers are provided by long-lived PC, independent of memory B cells and demonstrate that this humoral memory is inert to antigen. PMID- 9846700 TI - MHC class II and invariant chain biosynthesis and transport during maturation of human precursor dendritic cells. AB - Dendritic cells (DC) are highly potent activators of the immune response. The precise mechanisms that give rise to the DC phenotype are not known. To investigate the mechanisms that contribute to the generation of the DC phenotype, precursor DC were freshly isolated from human blood and allowed to mature in vitro. These matured DC showed the phenotypical and functional characteristics of DC. Analysis of the MHC class II and invariant chain (li) biosynthesis revealed that upon maturation, class II synthesis was induced whereas li synthesis was significantly up-regulated. In mature DC, despite the presence of large amounts of li, export of MHC class II molecules from the endoplasmic reticulum was incomplete, up to 4 h after biosynthesis. Thus, MHC class II-li synthesis and transport in DC is highly regulated during maturation of DC. Analysis of the regulatory mechanisms may contribute to a better understanding of antigen presenting capacities during the differentiation of DC. PMID- 9846701 TI - Conserved sequence motifs of CDR3 loops of TCR specific for two major epitopes of the grass pollen allergen PhI p 1. AB - We analyzed the cDNA sequence data of complementarity determining regions (CDR3) of epitope mapped alphabeta TCR of T cell clones (TCC). The TCC were specific for the major timothy grass (Phleum pratense) pollen allergen Phl p 1 and were derived from the peripheral blood of seven unrelated grass pollen-allergic individuals. Each TCR recognized one of two immunodominant T cell epitopes, PP73 or PP103, of Phl p 1. Although a diversity of recombined V and J segments was observed, amino acid motifs as long as five residues were conserved among CDR3 loops of TCR from TCC of different atopic individuals specific for the same peptide. The conserved sequences could comprise as much as 60% of the CDR3. All amino acid residues of the motifs of the CDR3beta and most of the CDR3alpha of all TCR used in this study were encoded by randomly added nucleotides. This indicates that they were specifically selected for by the peptide bound to the MHC class II molecule. For one selected patient, a larger number of TCC, specific for PP103, was analyzed. The TCR repertoire was limited to three different TCR. The same MHC class II molecule, DRB1*1301, was identified to present PP103 to each of the three TCR. PMID- 9846702 TI - Activation-induced apoptosis of mature T cells is dependent upon the level of surface TCR but not on the presence of the CD3 zeta ITAM. AB - Activation-induced cell death (AICD) occurs primarily in recently activated T cells after a second TCR triggering. Since a threshold in the activation status may be critical for AICD, it is likely that the CD3 ITAM, docking sites for tyrosine kinases, regulate AICD. A 'threshold model' for AICD was tested by using two targeted mutant mouse strains lacking either the zeta chain (CD3zeta-/-) or the ITAM of the zeta chain (CD3zeta-/-:Tgzetadelta67-150). Although the T cells from the CD3zeta-/- mice express extremely low levels of surface TCR, a subpopulation (approximately 18%) of activated T cells could be induced to express TCR/FcepsilonRI gamma by using a powerful polyclonal activation protocol. These activated TCR/FcRI gamma T cells were capable of undergoing AICD, but its induction required 10 times as much anti-CD3epsilon mAb as that required for AICD of wild-type T cells. Thus, the intensity of AICD correlated with the level of CD3 expression and was less efficient with activated, CD3zeta(-/-)-derived T cells. By contrast, AICD of T cells from the CD3zeta-/-:Tgzetadelta67-150 mice could be induced with low doses of anti-CD3epsilon mAb and the extent of AICD was comparable to T cells from wild-type mice. The AICD induced in T cells from CD3-/ , CD3zeta-/-:Tgzetadelta67-150 and normal controls was specifically inhibited by Fas-Ig fusion proteins. Our data support the 'threshold model' of AICD by demonstrating that AICD is controlled by the strength of T cell activation. PMID- 9846703 TI - Prevention of antigen-induced microtubule organizing center reorientation in cytotoxic T cells by modulation of protein kinase C activity. AB - Lysis of target cells (TC) by cytotoxic T lymphocytes (CTL) is achieved by directional exocytosis of cytolytic molecules-perforin and granzymes. They are stored within lytic granules which can be readily released following antigenic stimulation. Secretion of lytic molecules appears to be controlled by protein kinase C (PKC) activity, since specific modulators of PKC activity abolish the lysis of TC. We have examined the effect of PKC modulation on some of the earliest events in the perforin/granzyme-mediated cytotoxicity. De novo synthesis of perforin mRNA, required for the refilling of granules and sustained cytotoxicity, seems to be unaltered in the presence of PKC modulators. Immunofluorescent studies of CTL-TC conjugates revealed that PKC modulation impairs reorientation of the microtubule organizing center toward the contact point with the TC, which accounts for the specific direction of lytic granules exocytosis. Thus, it appears that PKC regulates exocytosis of lytic granules by governing microtubule reorganization, one of the initial steps in perforin/granzyme-mediated cytotoxicity. PMID- 9846704 TI - Defective IL-2-mediated IL-2 receptor alpha chain expression in Stat3-deficient T lymphocytes. AB - Stat3, a member of signal transducers and activators of transcription (STAT), is activated by a variety of cytokines. Recently, mice lacking Stat3 specifically in T cells have been generated and shown to be defective in IL-6-induced proliferation due to the impairment in IL-6-mediated prevention of apoptosis. In the present study, we show that Stat3-deficient T cells are partially defective in IL-2-induced proliferation. Stat3-deficient T cells show impaired IL-2 mediated IL-2 receptor (IL-2R) alpha chain expression. When Stat3-deficient T cells are stimulated with high-dose IL-2, these T cells express IL-2Ralpha and proliferate to similar extents as wild-type T cells. These demonstrate that Stat3 activation is required for efficient T cell proliferation by IL-2 through IL 2Ralpha induction. Taken together, these findings demonstrate that Stat3 activation in T cells is responsible for IL-2- and IL-6-induced proliferation through distinct mechanisms. PMID- 9846705 TI - Use of antibiotics during pregnancy. AB - OBJECTIVE: To describe the occurrence and distribution of antibiotic treatments, in addition their indications in control pregnant women in the population-based large dataset of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1996. RESULTS: Of 38 151 control pregnant women who delivered later newborn infants without congenital abnormality, 6554 (17.2%) were treated by antibiotics. Most women (14.5%) had penicillin, while 1.2% and 0.7% of pregnant women were treated by cephalosporins and tetracyclines, respectively. More than 100 pregnant women used the following antibiotics: ampicillin (6.9%), penamecillin (5.9%), cefalexin (1.0%), phenoxymethylpenicillin (0.6%), oxytetracycline (0.5%), erythromycin (0.45%), benzylpenicillin-procain (0.4%) and benzylpenicillin+benzylpenicillin-procain (0.3%). Different antibiotics had different indications for treatment. The mean birth weight was significantly lower in the treated group compared to the untreated group. PRACTICAL IMPLICATIONS: Different antibiotics have different chemical structures and indications for treatment. Therefore it is not appropriate to evaluate their teratogenic potential of combined antibiotic groups. There may be many interactions between underlying maternal diseases, other drug uses, further confounding factors and antibiotics studied, thus adequate controls are needed to estimate the adjusted teratogenic odds-risk ratios. European countries have different spectrum of antibiotic use. It would be necessary to know these baseline data of different populations. The anxiety and fear created by the notion that nearly all drugs cause congenital abnormalities may be more harmful than some proven human teratogenic drugs themselves. Thus a better risk-benefit estimation for antibiotic uses during pregnancy is an urgent and important task. PMID- 9846706 TI - Meconium stained amniotic fluid in preterm delivery is an independent risk factor for perinatal complications. AB - OBJECTIVE: To determine the prevalence and clinical significance of meconium stained amniotic fluid (MSAF) in women with preterm delivery. STUDY DESIGN: The study population consisted of consecutive patients who arrived with intact membranes and delivered preterm, singleton neonates at the Soroka Medical Center between 1 January 1985 and 31 December 1995. Only vertex presentation was included. Antepartum death was excluded from the study. Patients were classified according to the color of amniotic fluid into two groups: MSAF and clear amniotic fluid. Maternal puerperal complications were defined in our study as the presence of at least one of the next variables: clinical chorioamnionitis; major puerperal infection including endometritis, cesarean section or postpartum hemorrhage. Perinatal complications were defined in our study as: (1) intrapartum death (IPD) or postpartum death (PPD); (2) one or more of the following: 1-min Apgar score <3, 5-min Apgar score <7 or small for gestational age. Rates of perinatal complications were assessed at: (1) 24-27 weeks; (2) 28-31 weeks; (3) 32-36 weeks. Logistic regression was used to investigate the relationship of MSAF to perinatal complications and maternal morbidity in a multivariate model. RESULTS: During the study period, a total of 96 566 deliveries occurred in our institution and 4872 (5.0%) deliveries were preterm. Among the women delivering preterm meeting eligibility criteria, 276 (5.7%) women had intrapartum MSAF. A higher rate of IPD and PPD was observed only between 32 and 36 weeks' gestation in patients with MSAF in comparison with patients with clear amniotic fluid [6.1% (14/230) vs. 2.1% (85/4045), respectively, P=0.0001]. A statistically significant higher rate of perinatal complications was found between 28 and 31 weeks' gestation, and even a higher rate was noted between 32 and 36 weeks' gestation in the MSAF group in comparison with patients with clear amniotic fluid [51% (18/35) vs. 27.2% (93/341), respectively, P=0.003; 20% (46/230) vs. 9.8% (396/4045), respectively, P=0.0004]. CONCLUSIONS: (1) MSAF is an independent risk factor for perinatal complications in preterm deliveries (OR=1.73, CI: 1.057-2.43, P=0.001; OR=2.35, CI:1.34-4.12, P=0.002, respectively). (2) MSAF was not found to be an independent risk factor for maternal morbidity. PMID- 9846707 TI - A randomized comparison of vaginal prostaglandin E2 with oxytocin plus amniotomy for induction of labour in women with intermediately ripe cervices. AB - OBJECTIVE: To compare the effects of oxytocin and amniotomy or vaginal prostaglandin E2 (PGE2) for induction of labour. STUDY DESIGN: We conducted a randomized clinical trial. Eligible for the trial were women with normal pregnancy, parity 0-3, with intact membranes, >40 weeks of gestation documented by ultrasound examination before 20 weeks gestation, observed in a network of 13 general and teaching hospitals in Italy. Inclusion criteria were cervical Bishop's score 5-7, less than six uterine contractions per hour, single pregnancy, cephalic presentation, no history of cesarean section and uterine surgery. Eligible women were randomly assigned by phone to oxytocin plus amniotomy (163 women) or vaginal PGE2 2 mg, two doses at 6-h intervals (157 women). RESULTS: Overall, 50 women (15.6%) delivered by cesarean section, 22 (13.5%) randomized to oxytocin, and 28 (17.8%) randomized to PGE2 (not significant). Twelve hours after randomization, induction had failed in 26 women of the 163 randomized to oxytocin plus amniotomy (21.6%) and 34 out of the 157 randomized to PGE2 (15.9%): the difference was not significant. Neonatal outcome was similar in the two groups. CONCLUSIONS: This study did not find marked differences in labour and neonatal outcome between women randomized to oxytocin plus amniotomy or vaginal PGE2. A shorter induction delivery interval in the group receiving amniotomy and oxytocin after PGE2 priming was observed. PMID- 9846708 TI - Peripartum complications in grand multiparous women: para 6-9 versus para > or =10. AB - OBJECTIVE: To assess the importance of birth order and advanced maternal age on maternal and peripartum complications. STUDY DESIGN: The study population consisted of 12 296 multiparous women (six deliveries or more) with singleton gestation. Patients were classified into two groups according to the birth order: grand multiparous (between six and nine deliveries) and huge multiparous (ten or more deliveries). Peripartum complication was defined when at least one of the following conditions occurred: massive hemorrhage, uterine rupture, abruptio placentae, dysfunctional labor or malpresentations. Logistic regression analysis was used to evaluate the relationship between birth order and maternal age and peripartum complications. RESULTS: Among the study population, 9587 (78%) were grand multiparous and the remaining 2709 were huge multiparous women. The rate of peripartum complications was higher in huge multiparous than in grand multiparous women: malpresentation (6.2% versus 5%, P<.005), massive hemorrhage (0.7% versus 0.4%, P<.001) and dysfunctional labor (6.4% versus 3.5%, P<.001). Huge multiparous women also had a higher rate of the following complications than grand multiparous women: cesarean section (14.4% versus 10.4%, P<.01), chronic hypertension (7.9% versus 3%, P<.001), severe pregnancy induced hypertension (2.6% versus 1.1%, P<.01), diabetes class A (10.7% versus 7.5%, P<.005), diabetes class B-R (4.3% versus 2%, P<.01) congenital anomalies (3.3% versus 2.6%, P<.05) and large for gestational age infant, (17% versus 12.4%, P<.01). When adjusted for maternal age, high birth order remained strongly associated with the occurrence of peripartum complications. CONCLUSIONS: Huge multiparity was associated with a higher rate of maternal and peripartum complications than grand multiparity. Higher birth order remained an independent risk factor for peripartum complications after adjustment for maternal age. PMID- 9846709 TI - Recombinant human erythropoietin treatment of postpartum anemia. Preliminary results. AB - OBJECTIVES: The aim of this study was to investigate the efficacy of recombinant human erythropoietin (rHuEpo) in postpartum anemia. STUDY DESIGN: At the University Hospital of Ioannina, rHuEpo was administrated subcutaneously to twenty anemic women (hemoglobin [Hb]<10 g/dl), for 15 days following delivery; all were given iron and folic acid per os. Twenty other women (the control group) with postpartum anemia (Hb<10 g/dl), received only iron and folic acid. The Mann Whitney U-test was used for the comparison of hematological indices between the two groups, on days 1, 3, 5, 10, 15 and 40 postdelivery. RESULTS: On day 3, reticulocyte counts were significantly higher in the women who received rHuEpo, as compared to the controls (P<0.05). The mean Hb value increased to >2 g/dl in the group undergoing rHuEpo therapy as compared to 0.7 g/dl in the control group on day 5 (P<0.05). Furthermore, two women in the control group required blood transfusions, while no transfusions were required by the rHuEpo group. CONCLUSIONS: rHuEpo administration is useful for a more rapid amelioration of hematological indices in women with postpartum anemia. Further, the dose given in this study was not associated with significant side-effects. PMID- 9846710 TI - Prediction of patients with higher order multifetal pregnancy at risk for postpartum pulmonary edema. AB - OBJECTIVE: This retrospective study aims to verify the factors for the development of maternal pulmonary edema in higher order multifetal pregnancy. STUDY DESIGN: We analyzed medical profiles of a total of 13 triplet, quadruplet and quintuplet pregnancies for the years 1992 through 1997. Some treatments were applied in attempts to promote these multifetal pregnancies. All underwent cesarean section, two of which developed pulmonary edema within a few hours of delivery. There had been no evidence for the development of pulmonary edema antepartum. RESULTS: In the patients affected by pulmonary edema, postoperative values of PaO2/FIO2<250 mmHg showed close association to a value perioperative fluid loading index (FLI)>0; the index consists of an intraoperative fluid balance and preoperative infusion volume within 24 h prior to surgery. Two patients with postoperative pulmonary edema had a perioperative FLI>0, whereas the others had values 0 may have a much higher risk for postoperative pulmonary edema, suggesting the predictive role of the perioperative FLI value. PMID- 9846711 TI - Laparoscopic gonadectomy in male pseudohermaphrodites. AB - OBJECTIVE: To study patients of male pseudohermaphroditism and establish the laparoscopic approach for gonadectomy in these cases. STUDY DESIGN: Seven phenotypic females with XY karyotype were evaluated through a diagnostic protocol which included clinical, cytogenetic, hormonal, endoscopic and histologic evaluation. The gonads were then removed by laparoscopic surgery. RESULTS: The seven patients included three patients of pure gonadal dysgenesis, two patients of testicular feminization and one patient each of mixed gonadal dysgenesis and dysgenetic male pseudohermaphroditism. Two of the seven patients (28.57%) had gonadal neoplasias on histopathology-one dysgerminoma and one occult seminoma. In all of these patients, removal of the gonads was accomplished laparoscopically. No complications occurred during any of the surgeries. CONCLUSION: Due to the reduced morbidity, shorter hospital stay and safety, laparoscopic gonadectomy can be considered the treatment of choice for the removal of gonads in male pseudohermaphrodites in the hands of experienced laparoscopic surgeons. PMID- 9846712 TI - Depressive symptoms are often unrecognized in gynaecological practice. AB - We performed a study on depressive symptoms in gynaecological practice in Belgium, identifying depressive symptoms according to both practitioners' opinions and the scores on a scale developed to detect depression in general medical settings. According to the 170 participating gynaecologists, 12% of the 2174 women they interviewed had symptoms of depression. However, 35% had two or more positive answers on the Depression Scale, and 19% had four or more positive answers. Our results suggest that depressive symptoms are often not identified in clinical gynaecological practice. PMID- 9846713 TI - Surgical treatment of ovarian dermoid cysts. AB - OBJECTIVE: To evaluate the efficacy of laparoscopic ovarian cystectomy and to compare the surgical course, post-surgical course and particularly post-surgical pain of the laparoscopic and laparotomic methods. STUDY DESIGN: We conducted a surgical study on dermoid cysts at the Gynecology Department of Siena University between 1 January 1992 and 31 December 1996. The selected cases were randomized into two groups based on surgical approach: via laparotomy (n=22) or laparoscopy (n=22). Surgical times, estimated blood loss, post-surgical pain, time in hospital, speed of recovery and complications were compared. RESULTS: Mean blood loss was significantly less for laparoscopy (58.64+/-30.17 ml versus 103.84+/ 38.45 ml, P<0.05). Mean hospitalization was 6.32+/-1.09 days for laparotomy and 3.18+/-0.39 days for laparoscopy (P<0.05). Post-surgical pain was significantly less in laparoscopy patients (P<0.05). The laparoscopic technique had fewer post surgical complications. CONCLUSIONS: The laparoscopic approach had many advantages. Laparoscopy should be the elective treatment for women with dermoid cysts, because it has many advantages for the patient and lower costs for the national health system. PMID- 9846714 TI - High prevalence of cervical intra-epithelial neoplasia in women treated for pelvic inflammatory disease. AB - Because human papillomavirus (HPV) is sexually transmitted, as are pelvic inflammatory disease (PID) agents, the authors investigated whether cervical intra-epithelial neoplasia (CIN) are more frequent in women under treatment for PID. The study involved 298 patients hospitalized for PID, in whom CIN were investigated by smears and colposcopy. CIN were diagnosed in 42 patients, i.e. in 14% of patients: 21 low-grade CIN and 21 high-grade CIN, including one case of early-stage microinvasion. These figures are to be compared to the 0.5-4% of pre cancerous lesions found in the general population. Screening smears are frequently inaccurate and direct colposcopy appeared preferable. No clinical study of this type has been published before, but certain authors have mentioned a high CIN incidence in patients with PID histories or followed up in sexually transmitted disease centers. These results show that CIN are more frequent in patients treated for PID; CIN should be investigated systematically in this population. PMID- 9846715 TI - An objective experimental assessment of the learning curve for laparoscopic surgery: the example of pelvic and para-aortic lymph node dissection. AB - OBJECTIVES: To date the number of procedures required to become competent to perform new laparoscopic surgical techniques is not known. STUDY DESIGN: The pig model was chosen for assessment of the learning curve associated with an advanced laparoscopic procedure. A unilateral laparoscopic pelvic lymphadenectomy was performed by two residents and a laparoscopic para-aortic lymphadenectomy was performed by a fellow on a series of 20 pigs. The quality of the dissection was checked by immediate laparotomy by an independent observer. RESULTS: The operative objectives were: (a) There should be less than 5% residual lymph nodes. (b) The operating time should be less than 30 min for pelvic and less than 100 min for para-aortic lymphadenectomy. (c) Avoiding conversion because of complications. This target was achieved after 7 and 9 pigs respectively for pelvic lymphadenectomy and after 14 pigs for para-aortic lymphadenectomy. CONCLUSION: It is feasible to assess the learning curve of trainee surgeons while performing laparoscopic pelvic and para-aortic lymphadenectomy on pigs. A training programme such as this should prevent complications due to inexperience and should satisfy ethical and medico-legal considerations. PMID- 9846716 TI - Complications of abortion performed under local anesthesia. AB - OBJECTIVE: To assess the incidence of complications of abortion performed under local anesthesia. DESIGN: Prospective study. SETTING: A family planning center in the Paris area. POPULATION: Eight hundred and fifty-eight women admitted for abortion under local anesthesia. MAIN OUTCOME MEASURES: Incidence of immediate (the day of vacuum aspiration) and delayed complications (at the follow-up visit 2 weeks after the procedure). RESULTS: Among the 858 women who underwent vacuum aspiration, 683 (80%) attended the follow-up visit 2 weeks after the procedure. Fifty-nine percent of the 858 women had not had a previous abortion, 25% had had one, and 16% had had two or more. The average duration of amenorrhea was 8.6 weeks (19% at 6 or 7 weeks, 67% between 8 and 10 weeks, and 14% after 10 weeks). The overall complication rate was 3.4% (23/683) (95% CI=2.0-4.8%). The incidence of immediate complications was 1.7% (15/858) (0.8-2.6%) and that of delayed complications 1.2% (8/683) (0.4-2.0%). CONCLUSION: This study confirms the safety and efficacy of abortion by vacuum aspiration under local anesthesia. PMID- 9846717 TI - External cephalic version after previous cesarean section: a series of 38 cases. AB - OBJECTIVE: To determine if external cephalic version (ECV) is a reasonable alternative to repeat cesarean section in case of breech presentation. STUDY DESIGN: Retrospective study of 38 women with one previous cesarean section and a breech presentation after 36 weeks of gestational age who have had at least one experience of ECV. Statistics used the Fisher's test with significance when P<0.05. RESULTS: Version attempts were successful in 25 of the 38 women (65.8%). Seventy-six percent of the successful version women went on to have vaginal birth after cesarean section. A total of 19 successful vaginal deliveries occurred (50%). Success rate of ECV was lowered when breech was the indication of the previous cesarean section. The vaginal delivery rate was increased after successful ECV in patients previously vaginally delivered, but this difference did not reached significance (P=0.057). No maternal or neonatal complications occurred. CONCLUSION: ECV is acceptable and effective in women with a prior low transverse uterine scar, when safety criteria are observed. PMID- 9846718 TI - Cervical dysplasia and human immunodeficiency virus infection in women: prevalence and associated factors. Groupe d'Epidemiologie Clinique du SIDA en Aquitaine (GESCA). AB - OBJECTIVE: To study the prevalence of Cervical Intraepithelial Neoplasia (CIN) and its association with HIV-I infection, controlling for other risk factors of CIN. DESIGN: Cross-sectional survey. SUBJECTS AND METHODS: HIV-1 seropositive (HIV+) and seronegative (HIV-) women were enrolled at the Obstetrics-Gynecologic Departments of the Bordeaux University Hospital from April 1993 to June 1995. A gynecologic check-up was performed with a clinical examination, a colposcopy and a Papanicolaou smear. Sexually Transmitted Diseases (STDs) were screened. Colposcopy was interpreted as: normal, low-grade or high-grade lesions. Interpretation of Papanicolaou smears was based on the 1988 Bethesda system using three descriptive diagnoses: normal, low-grade and high-grade Squamous Intraepithelial lesions (SILs). If colposcopy showed a high grade lesion or Papanicolaou smear a high-grade SIL, a cervical biopsy was performed. Absence of CIN was defined by normal Papanicolaou smear and colposcopy. High grade CIN was defined by either identification of high grade SIL on Papanicolaou smear or high grade lesion on colposcopy confirmed by CIN2-3 lesion on biopsy. Other cases were classified as low-grade CIN. HPV infection was diagnosed on presence of koilocytosis on cytological or histological specimens. RESULTS: Prevalence of CIN was significantly higher in the 128 HIV+ women than in the 102 HIV- women: 34.4% vs. 13.7% (O.R.=3.30). Among HIV+ women, 25.8% had low-grade CIN and 8.6% high grade CIN versus 10.8% and 2.9%, respectively among the HIV- women. Prevalence of HPV infection was 50.0% among women with CIN vs. 5.8% in women without CIN (P= 10(-6)). In the multivariate analysis of the determinants of CIN, smoking more than ten pack-years and HPV infection were the only two variables associated with CIN while association with HIV infection disappeared. Among HIV+ women, the variables associated with CIN were clinical AIDS and HPV infection. PMID- 9846719 TI - Parasitic ovarian dermoid tumor of the omentum-A review of the literature and report of two new cases. AB - Two cases of a parasitic omental teratoma which originated from an ovarian dermoid that underwent torsion, autoamputation and omental reimplantation are presented. A review of the literature revealed 23 additional cases of omental teratoma which occurred mostly in females. In some cases, the mature teratoma of the omentum showed histological evidence of ovarian stroma, and was associated with a dermoid tumor of the remaining contralateral ovary. It is generally believed that autoamputation and reimplantation of an ovarian dermoid cyst is the most common etiology of omental teratomas. Abdominal pain is the main presenting symptom of these tumors, and on physical examination a mobile abdominal or pelvic mass is often found. Both ultrasonography with colour flow Doppler and CT-scan are helpful in the diagnosis of dermoid tumors, but the correct diagnosis of omental localisation is extremely difficult. Mature omental teratomas may be treated by simple resection. The immature teratomas of the greater omentum, however, are potentially malignant tumors requiring postoperative chemotherapy and radiotherapy. PMID- 9846720 TI - Long-term disease-free survival following surgery and active specific immunotherapy with allogeneic vaccine in a patient with high-risk malignant melanoma of the vulva. AB - Vulvar malignant melanoma with deep vertical penetration of the tumor and involvement of regional lymph nodes carries a very poor prognosis. The case of a 25-year-old woman with a history of a Breslow depth 6.0 mm and Clark Level IV primary vulvar malignant melanoma, involving the anterior part of the left labium major, 1 cm from the clitoris, is reported. The patient had undergone a left radical hemivulvectomy and bilateral groin dissection. There were two of thirteen superficial left groin nodes containing metastatic melanoma. The patient had been treated postoperatively with an allogeneic specific anti-melanoma vaccine in combination with high-dose cimetidine and has survived without disease for more than five years. To the best of our knowledge this is the first case report in the literature of active specific immunotherapy with allogeneic vaccine in vulvar malignant melanoma. This case illustrates that the behavior of malignant melanoma, including vulvar malignant melanoma, is unpredictable and active specific immunotherapy with allogeneic vaccine may have a role in the postoperative treatment of high risk vulvar malignant melanoma. PMID- 9846721 TI - Vasodilator response to exogenous adrenocorticotropic hormone in fetal adrenal cortex precedes increased steroidogenesis in sheep at 105-112 days gestation. AB - OBJECTIVE: To examine whether the increase in adrenal cortical blood flow induced by adrenocorticotropic hormone (ACTH) is linked to steroidogenesis, or if these effects can be separated. STUDY DESIGN: Adrenal cortical responses to ACTH were measured in ovine fetuses at 105-112 days gestation, when the adrenal is hyporesponsive. Fetuses were given an intravenous infusion of ACTH(1-24) or of vehicle. We measured regional adrenal blood flows by the microsphere technique and plasma ACTH and cortisol levels by radioimmunoassay. RESULTS: After 3 and 24 h of ACTH infusion, plasma cortisol concentrations had risen from 2.8+/-0.8 ng/ml to 8.4+/-0.3 and 78.0+/-15.8 ng/ml (means+/-S.E.M.). Adrenal cortical blood flow increased from 108+/-25 ml/min/100 g to 319+/-33 and 518+/-51 ml/min/100 g. Plasma cortisol levels and adrenal blood flow did not change in control fetuses. CONCLUSION: Since there was a rapid blood flow response to ACTH, despite the expected delay in the rise of plasma cortisol, we suggest that vascular responses to ACTH are not tightly linked to adrenal metabolic activity in the ovine fetus at 0.7 gestation. PMID- 9846722 TI - Leptin levels and gene expression during the perinatal phase in the rat. AB - The role of leptin in controlling food intake and adiposity has been the aim of many different investigations in the last 3 years. Pregnancy and lactation are two physiological situations associated with a clear hyperphagia (together with important changes in metabolism and adipose mass) to sustain the different and varying demands for foetal growth and milk production respectively. We therefore focused on the role of leptin in perinatal hyperphagia. The circulating leptin levels and leptin gene expression in adipose tissue of both pregnant and lactating rats were examined. Pregnant rats showed unchanged adipose tissue leptin mRNA levels but increased circulating leptin; this probably reflects the high fat carcass content characteristic of pregnancy. Conversely, lactating rats did not show any change either in circulating leptin or adipose tissue mRNA levels. Litter-removal caused a significant increase in both circulating leptin levels and gene expression. The results obtained permit us to suggest that leptin does not seem to have a role in controlling food intake during the perinatal phase. PMID- 9846723 TI - Thinning of the intervascular tissue layers of the human placenta is an adaptive response to passive diffusion in vivo and may help to predict the origins of fetal hypoxia. AB - OBJECTIVES: To test the hypothesis that thinning of the placental intervascular layers, and greater variability in thickness, are positive adaptations to facilitate passive diffusion and may help to resolve different categories of fetal hypoxia. STUDY DESIGN: Placentas from 12-41 weeks of normal gestation and from pregnancies associated with fetal hypoxic stress (high altitude, diabetes mellitus) were sampled systematically, fixed in formalin, wax-embedded and quantified using stereological methods. Arithmetic and harmonic mean distances across villous trophoblast, stroma and fetal plasma were estimated by measuring randomly sampled intercept lengths. In each case, an index of variability of layer thickness was calculated by dividing the arithmetic by harmonic mean distance. This index has the value I when a tissue layer is uniformly thick but increases in value as local layer thickness becomes more variable. Comparisons between groups were drawn using variance and regression analysis. RESULTS: During pregnancy, there were significant negative correlations between layer thickness (trophoblast, stroma) and gestational age and fetal weight, and significant positive correlations between thickness irregularity and age and weight. Compared with lowland controls, high-altitude placentas possessed thinner layers but only the trophoblast and stroma (and not fetal plasma) were more variably thick. In maternal diabetes, only fetal plasma distance was reduced but fetal and stromal layers appeared to be more irregular in thickness. CONCLUSIONS: Alterations in placental intervascular layer thicknesses occur in normal and abnormal pregnancies and represent real adaptations leading to improved diffusive conductances. Differences in the tissue location of the adaptive response may depend on the nature and origins of the fetal hypoxia. PMID- 9846724 TI - Aggressive angiomyxoma of the vulva--a case report. PMID- 9846725 TI - Case of acrania associated with congenital medulloblastoma. AB - A case of acrania associated with medulloblastoma, agenesis of the cerebellum, and nasoshizis, is reported. The diagnosis of acrania was made at the 20th gestational week by sonographic examination. To our knowledge, this is the first report of fetal acrania associated with congenital brain tumor. PMID- 9846726 TI - Intracranial subdural haematoma complicates accidental dural tap during labour. AB - A 19-year-old, healthy nulliparous woman developed an intracranial subdural haematoma after unintentional dural puncture in connection with epidural analgesia for labour pain. The haematoma was evacuated and the patient recovered completely. Anaesthesiologists and obstetricians should be aware of this rare but potentially dangerous complication to epidural analgesia. PMID- 9846727 TI - Ovarian dysgerminoma showing high serum levels and positive immunostaining of placental alkaline phosphatase and neuron-specific enolase associated with elevation of serum prolactin level. AB - We report a 35-year-old Japanese female patient with ovarian dysgerminoma showing elevated serum levels of placental alkaline phosphatase (PLAP), neuron-specific enolase (NSE) and prolactin (PRL). All elevated tumor markers improved dramatically after the removal of the tumor. Immunohistochemically examined, the tumor was stained positive for PLAP and NSE and negative for PRL. Our present case is the first report of dysgerminoma showing positive immunostaining for PLAP and NSE, and the association of high serum level of PRL followed by decrease accompanied by the tumor debulking. PMID- 9846728 TI - Helicobacter pylori--a conundrum of genetic diversity. PMID- 9846729 TI - Candida albicans SSD1 can suppress multiple mutations in Saccharomyces cerevisiae. AB - The SSD1 gene of Saccharomyces encodes a 160 kDa cytoplasmic protein that can suppress mutations in a number of other genes. A functional homologue of SSD1 from the human pathogen Candida albicans was isolated on the basis of its ability to restore viability at the restrictive temperature in a Saccharomyces cerevisiae swi4 ssd1-d strain. The C. albicans gene, designated CaSSD1, encodes a 1262 aa protein which has 47% identity overall to S. cerevisiae SSD1 as well as significant identity to Schizosaccharomyces pombe dis3 and sts5 products. It is shown that CaSSD1 expression is constitutive through the mitotic cell cycle, which is consistent with a role for the protein in cell growth. CaSSD1 rescues the swi4ts defect in an ssd1-d background when expressed from its own promoter on a single-copy plasmid and under the same conditions can rescue mutations in genes encoding protein phosphatase type 2A catalytic subunits. These data suggest that CaSSD1, like its S. cerevisiae homologue, can limit the effect of mutations on a variety of cellular processes. PMID- 9846730 TI - A Candida albicans chaperonin subunit (CaCct8p) as a suppressor of morphogenesis and Ras phenotypes in C. albicans and Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae and the pathogen Candida albicans can be induced to undergo morphogenesis from a yeast to a filamentous form. A C. albicans gene (CaCCT8) was identified encoding a subunit of the Cct chaperonin complex, whose expression prevents filament formation in both fungi without interfering with growth of the yeast form. In S. cerevisiae, pseudohyphal growth induced by Ras2Val19, by overproduction of Phd1p or by expression of the C. albicans EFG1 gene, was blocked by CaCct8p and its N-terminally deleted derivative CaCct8 delta1p; in contrast, pseudohyphal induction by other components (Cph1p, Cdc42p) could not be suppressed, indicating that morphogenesis per se is not inhibited. CaCCT8 expression also interfered with other Ras2pVal19 phenotypes, including heat sensitivity, lack of glycogen accumulation and lack of sporulation. In C. albicans, overproduction of CaCct8p effectively blocked hyphal morphogenesis induced by starvation conditions and by serum. The results suggest that the activity of a component in the Ras2p signal transduction pathway is suppressed by excess chaperonin subunits. This component may be a novel folding target for the Cct complex. In agreement with this hypothesis, disruption of one of the two CaCCT8 alleles in C. albicans led to defective hyphal morphogenesis. PMID- 9846731 TI - Identification of three differentially expressed hydrophobins in Pleurotus ostreatus (oyster mushroom). AB - Three proteins with characteristic features of class I hydrophobins, designated POH1, POH2 and POH3, were isolated from the basidiomycete Pleurotus ostreatus. Based on N-terminal sequence analyses, their cDNAs were isolated using RT-PCR; the cDNAs and corresponding genes were sequenced and their regulation studied. POH1 is expressed in the fruiting bodies but not in vegetative mycelium. The regulation of POH2 and POH3 is tightly correlated. Both genes are switched off in the fruiting bodies but abundantly expressed in the vegetative mycelium of both monokaryon and dikaryon. POH2 and POH3 were isolated from the culture medium and from aerial hyphae. Co-purified POH2 and POH3 assembled in vitro into a protein membrane with a typical rodlet pattern as found previously with other hydrophobins. Similar structures were detected on the surface of aerial hyphae. PMID- 9846732 TI - Molecular and biochemical analysis of a 105 kDa Mycoplasma gallisepticum cytadhesin (GapA). AB - The identification of a gene (gapA) from Mycoplasma gallisepticum with homology to the P1 cytadherence gene of Mycoplasma pneumoniae is reported. The gapA gene is a 2895 bp ORF encoding a protein with a molecular mass of 105 kDa. Nucleotide sequence analysis of the gapA gene revealed 45% homology to the M. pneumoniae P1 gene, 46% homology to the Mycoplasma genitalium MgPa gene and 47% homology to the Mycoplasma pirum P1-like protein gene. It has a 64 mol % A+T content compared to 46, 60 and 72 mol % respectively for the P1, MgPa and the P1-like protein genes. As with the P1 and MgPa genes, gapA is a central gene in a multi-gene operon, but unlike the P1 and MgPa genes, there is only a single copy of gapA in the genome. GapA is a trypsin-sensitive surface-exposed protein. Chicken tracheal-ring inhibition-of-attachment assays, using anti-GapA Fab fragments, resulted in 64% inhibition of attachment. These results indicated that GapA plays a role in cytadherence of M. gallisepticum to host cells. PMID- 9846733 TI - Truncation as a novel form of variation of the p50 gene in Mycoplasma hominis. AB - A characteristic feature of the mycoplasmas is the presence of variable surface proteins which may play an important role in the adaptation of the cell-wall-less organisms to their host environments. In addition, this antigen variation may be an important pathogenic property of the organism. The ubiquity of the gene encoding P50, an adhesin of Mycoplasma hominis FBG, and its transcription were analysed in different isolates of M. hominis. The p50 gene was present in all isolates tested. Based on Southern blot analysis and sequencing of the gene, the isolates could be classified into one of three distinct groups. Within two groups specific truncations of the p50 gene occurred. The reduction of the gene size was confirmed in Northern blot analysis of representative isolates from each group, with a decrease in transcript length from 1.6 kb in group G-1 down to 0.76 kb in group G-3. In addition to truncation, a coincidental duplication of some gene segments was detected. This work has provided evidence for the genetic basis of a further variation in the M. hominis P50 adhesin. PMID- 9846734 TI - Internalization and cytotoxicity are important virulence mechanisms in Vibrio fish epithelial cell interactions. AB - Vibrio anguillarum and Vibrio damselae are Gram-negative bacteria that cause systemic infections called vibriosis in fish. They can enter fish cells and survive as intracellular parasites. The host-pathogen interactions between these Vibrio species and the fish epithelial cell lines epithelioma papillosum of carp (EPC) and grunt-fin tissue (GF) cells, were examined using phase-contrast, scanning electron and confocal microscopy. In addition, potential signal transduction pathways that precede bacterial internalization were studied by using signal transduction inhibitors. Some Vibrio species induced morphological changes in fish cells and this allowed classification into a cytopathic group and a noncytopathic group. The cytopathic group could be subdivided into two invasive groups (I and II) and a cytotoxic group. Of the invasive strains V. anguillarum 811218-5W (group I) and G/Virus/5(3) (group II), genistein, a tyrosine kinase inhibitor, only inhibited internalization of V. anguillarum G/Virus/5(3) into EPC cells, whereas staurosporine, a protein kinase C inhibitor, accelerated internalization of both strains. Cytochalasin D, an inhibitor of microfilament polymerization, prevented internalization of both strains, whilst vincristin, a microtubule inhibitor, only inhibited internalization of V. anguillarum G/Virus/5(3). For the cytotoxic strain V. damselae ATCC 33539, extracellular products (ECP) alone caused morphological changes in EPC and GF. Bacterial internalization may not be important in the pathogenesis of this group. The non cytopathic strain V. anguillarum S2/5/93(2) did not enter cells or induce any changes in EPC and GF monolayers. This study has identified some major differences between Vibrio species in their interactions with fish cells in vitro and will thus facilitate future studies of the molecular basis of pathogenesis of vibriosis. PMID- 9846735 TI - Porin alteration and active efflux: two in vivo drug resistance strategies used by Enterobacter aerogenes. AB - Enterobacter aerogenes is among the five most frequently isolated nosocomial pathogens in France, and this bacterium also shows increasing multidrug resistance. In this study, various E. aerogenes strains isolated from hospital units were characterized for their outer-membrane proteins, antibiotic susceptibilities (inhibition diameters and MICs) and resistance mechanisms associated with modification of envelope permeability (porin alteration and active efflux). Diminished outer-membrane permeability due to porin alterations was found in conjunction with the expression of an enzymic barrier in resistant isolates. Interestingly, changes in the functional expression of porins appeared to play a special role in susceptibility to cefepime. An active efflux to quinolones was also identified. Simultaneous changes in envelope permeability, i.e. a porin deficiency (in) and an efflux mechanism (out), were clearly evident in two clinical strains. PMID- 9846736 TI - Characterization of Trichomonas vaginalis AP33 adhesin and cell surface interactive domains. AB - Adherence to host target cells is a critical step in establishing infection with the sexually transmitted pathogen Trichomonas vaginalis. Four parasite surface proteins mediating attachment to vaginal epithelial cells have been identified. One surface protein, termed AP33, was characterized further to identify domains interactive with previously generated antibodies and with host surface sites. N- and C-terminal deletion subclones were generated and tested for reactivity with both mAb and rabbit antiserum against AP33, and were also examined for their ability to bind to host cells. Surprisingly, the rabbit antiserum known to inhibit cytoadherence recognized an epitope(s) contained within 72 residues in the N-terminal half of the protein. However, the mAb epitope was immunoreactive with a 28-amino-acid region near the C-terminus. Subsequent mapping of this region with overlapping peptides identified a nine-amino-acid sequence reactive with the mAb. Equally surprising, two domains interactive with host cell surfaces were identified at distinct parts of AP33: one in the N-terminal half of the protein, and the other within 24 residues in the C-terminal third. Further analysis of the C-terminal binding domain revealed that a peptide representing this area could inhibit T. vaginalis cytoadherence by 40%. PMID- 9846737 TI - A lipid A-associated protein of Porphyromonas gingivalis, derived from the haemagglutinating domain of the RI protease gene family, is a potent stimulator of interleukin 6 synthesis. AB - There is evidence that the lipid A-associated proteins (LAPs) of enteric bacteria can induce the synthesis of interleukin 1 (IL-1) and therefore may be important virulence factors. Porphyromonas gingivalis is now recognized as a major pathogen in the chronic inflammatory periodontal diseases and it has previously been reported that a crude LAP fraction from this organism could induce IL-1 and interleukin 6 (IL-6) synthesis. In the present study the chemical and biological properties of the LAPs of this bacterium have been further characterized. Analysis by SDS-PAGE has shown that the LAPs comprise nine proteins of molecular masses 81, 68, 48, 47, 28, 25, 20, 17 and 16 kDa. These LAPs, at concentrations as low as 100 ng ml(-1), were shown to stimulate human gingival fibroblasts, human peripheral blood mononuclear cells and whole human blood to produce the pro inflammatory cytokine IL-6. The cytokine-inducing activity of the LAPs was reduced after heat-inactivation and trypsinization, suggesting that the activity was not due to contaminating LPS. To establish which proteins in this mixture were the active cytokine inducers, the LAPs were separated by electrophoresis on polyacrylamide gels. The majority of the activity was associated with the 17 kDa LAP. N-terminal sequence analysis demonstrated that this protein was homologous to an internal region of a conserved adhesin domain contained within a family of P. gingivalis extracellular proteins including the RI protease, Lys-gingipain, porphypain and haemagglutinin A. In addition to a role in adherence, the adhesin domain(s) of these proteins may also have cytokine-inducing properties. Furthermore, it has also been shown that the previously observed degradation of cytokines by P. gingivalis may be attributable to the catalytic domain of the RI protease. Thus, different domains within the same molecule appear to have opposing actions on pro-inflammatory cytokine levels and the balance between these two activities may influence the cytokine status of the periodontium in patients with the common chronic inflammatory conditions known as the periodontal diseases. PMID- 9846738 TI - Immunization with recombinant class 1 outer-membrane protein from Neisseria meningitidis: influence of liposomes and adjuvants on antibody avidity, recognition of native protein and the induction of a bactericidal immune response against meningococci. AB - The porA gene from Neisseria meningitidis was cloned into the pRSETA vector and recombinant class 1 outer-membrane protein expressed at high levels in Escherichia coli. The protein was readily purified by affinity chromatography on a Ni2+ matrix and used for immunization of mice with conventional AI(OH)3 adjuvant, with experimental adjuvants which have the potential for human use, and with liposomes. The resulting sera were analysed for the magnitude, subclass distribution and antigenic specificity of the immune response. In addition, surface plasmon resonance (SPR) was used to quantify antibody avidity by analysis of the kinetics of binding to native class 1 protein. Immunization with conventional and experimental adjuvants induced antibodies of low avidity that did not recognize native class 1 protein. In contrast, immunization with recombinant protein in liposomes induced antibodies of high avidity which recognized native class 1 protein, as measured by their ability to label meningococcal cells in immunofluorescence assays and to inhibit the binding of a protective mAb. These properties were associated with the presence in sera of high levels of antibodies with the ability to induce complement-mediated killing of meningococci. These data show that liposomes containing recombinant class 1 protein represent a potential basis of future vaccines, of defined composition, designed for the prevention of group B meningococcal infections. PMID- 9846739 TI - Biofilm susceptibility to bacteriophage attack: the role of phage-borne polysaccharide depolymerase. AB - Biofilm bacteria Enterobacter agglomerans 53b and Serratia marcescens Serr were isolated from a food processing factory. A bacteriophage (SF153b), which could infect and lyse strain 53b, was isolated from sewage. This has been shown to possess a polysaccharide depolymerase enzyme specific for the exopolysaccharide (EPS) of strain 53b. Using batch culture and chemostat-linked Modified Robbins Device systems it was observed that SF153b could degrade the EPS of a mono species biofilm (strain 53b) and infect the cells. The disruption of the biofilm by phage was a combination of EPS degradation by the depolymerase and infection and subsequent cell lysis by the phage. Strain Serr biofilms were not susceptible to the phage and the biofilm EPS was not degraded by the phage glycanase, with the result that the biofilm was unaffected by the addition of SF153b phage. Scanning electron microscopy confirmed that specific phage could extensively degrade susceptible biofilms and continue to infect biofilm bacteria whilst EPS degradation was occurring. PMID- 9846740 TI - A multilocus sequence typing scheme for Streptococcus pneumoniae: identification of clones associated with serious invasive disease. AB - The population biology of Streptococcus pneumoniae is poorly understood. Most of the important issues could be addressed by the molecular characterization of large, well sampled populations from carriage and from the different manifestations of pneumococcal disease. The authors have therefore developed a pneumococcal multilocus sequence typing scheme and database by sequencing approximately 450 bp fragments of seven housekeeping loci from 295 isolates. The combination of alleles at the seven loci provided an allelic profile, or sequence type (ST), and the relatedness between isolates was obtained by constructing a dendrogram from the matrix of pairwise differences between STs. The typing scheme was validated using pneumococci of known genetic relatedness and could resolve >6 billion STs. Among 274 isolates from recent cases of invasive pneumococcal disease in eight countries, 143 STs were resolved, but 12 STs contained at least five isolates (range 5-21 isolates). The repeated recovery of indistinguishable isolates from invasive disease in different countries implies that these STs define strains with an increased capacity to cause invasive disease. The relationship between STs and serotypes suggested that, in the longer term, capsular genes have been distributed horizontally within the pneumococcal population, but in the short term, expansion of clones occurs with only occasional changes of serotype. The multilocus sequence typing scheme provides a powerful new approach to the characterization of pneumococci, since it provides molecular typing data that are electronically portable between laboratories, and which can be used to probe aspects of the population and evolutionary biology of these organisms. A Web site for the molecular characterization of pneumococci by MLST is available (http ://mlst.zoo.ox.ac.uk). PMID- 9846741 TI - Electrotransformation and natural transformation of Streptococcus pneumoniae: requirement of DNA processing for recombination. AB - Electrotransformation has been used as a tool to introduce genes carried on replicative vectors in hundreds of bacterial species. In this study, the technique was used to try to obtain recombination of markers in the chromosome of the naturally transformable bacterium Streptococcus pneumoniae. Recombination was not observed even using naturally competent cultures. Both chromosomal and cloned DNA, denatured or native, were without effect. These results suggest that it is not sufficient to introduce DNA into the cell to obtain recombinants in this bacterium. The integration of markers into the chromosome in naturally competent cells must require DNA processing during entry. Electrotransformation of replicating plasmids is recA-independent but can be facilitated by a recA dependent process. This facilitation required the induction of the natural competence machinery, probably involving partial homologous pairing. PMID- 9846742 TI - Unconventional organization of the division and cell wall gene cluster of Streptococcus pneumoniae. AB - The genes responsible for cell wall biosynthesis and cell division (dcw genes) were identified and sequenced in Streptococcus pneumoniae. The genetic organization of the dcw cluster in Streptococcus pneumoniae differed significantly from the clusters of other bacteria reported to date. In particular, the genes corresponding to the 2 min region of the Escherichia coli chromosome were found distributed in three genetically separate regions of the Streptococcus pneumoniae chromosome. The first region contained the expected ftsA and ftsZ cell division genes at one end and pbp2b, ddl and murF at the other end. The murD, murG and divIB genes, always found located upstream of ftsA, were found in a second region separated from the first. A third region contained the yllC, yllD, pbp2x and mraY genes. The chromosomal region downstream of ftsZ was also sequenced and characterized. In Streptococcus pneumoniae this region contains four ORFs, all of unknown function, and an ORF encoding the Bacillus subtilis DivIVA homologue. The gene order and the organization of this region was found to be conserved in Staphylococcus aureus, Streptococcus pyogenes and Bacillus subtilis, raising the possibility that previously unidentified loci may also be involved in division. PMID- 9846743 TI - Streptococcus pyogenes protein F promotes invasion of HeLa cells. AB - Although the Gram-positive bacterium Streptococcus pyogenes (group A streptococcus) has been considered an extracellular pathogen which adheres to human mucosal epithelium, the streptococcus possesses invasive capacity for cultured human epithelial cells. This study provides genetic and functional evidence supporting the conclusion that protein F is capable of mediating entry of S. pyogenes into HeLa cells. Using Tn916 insertion mutagenesis or an isogenic S. pyogenes strain with a defined mutation in the gene encoding protein F (prtF), it was observed that the invasive capacity was affected by the levels of surface exposed protein F, but not by those of M protein. In addition, heterologous expression of protein F on Enterococcus faecalis conferred upon the bacteria an efficient invasive phenotype. Several assays demonstrated that both the fibronectin-binding domains of protein F, UR and RD2, were involved in host-cell invasion. In addition, coinfection experiments of HeLa cells with S. pyogenes and an Escherichia coli K-12 strain expressing an afimbrial adhesin AFA-I showed that the uptake of S. pyogenes did not permit internalization of the E. coli cells. PMID- 9846744 TI - Construction of a physical and preliminary genetic map of Aeromonas hydrophila JMP636. AB - A physical and preliminary genetic map of the Aeromonas hydrophila JMP636 chromosome has been constructed. The topology of the genome was predicted to be circular as chromosomal DNA did not migrate from the origin during PFGE unless linearized by S1 nuclease. Cleavage of the chromosome with PacI and PmeI produced 23 and 14 fragments, respectively, and enabled calculation of the genome size at 4.5 Mb. Digestion of the chromosome with I-CeuI produced 10 fragments, indicating that 10 rrl (23S) genes were likely to be present. Hybridizations between DNA fragments generated with PacI, PmeI and I-CeuI were used to initially determine the relationship between these segments. To accurately map genes previously characterized from JMP636, the suicide vector pJP5603 was modified to introduce restriction sites for PacI and PmeI, producing pJP9540. Following cloning of genes into this vector and recombinational insertion into the JMP636 chromosome, PacI and PmeI cleavage determined the location of genes within macrorestriction fragments with the additional bands produced forming hybridization probes. From the data generated, it was possible to form a physical map comprising all the fragments produced by PacI and PmeI, and assign the contig of I-CeuI fragments on this map. The preliminary genetic map defines the location of six loci for degradative enzymes previously characterized from JMP636, while the locations of the 10 sets of ribosomal genes were assigned with less accuracy from hybridization data. PMID- 9846745 TI - A vector for systematic gene inactivation in Bacillus subtilis. AB - To study the functions of the uncharacterized open reading frames identified in the Bacillus subtilis genome, several vectors were constructed to perform insertional mutagenesis in the chromosome. All the pMUTIN plasmids carry a lacZ reporter gene and an inducible Pspac promoter, which is tightly regulated and can be induced about 1000-fold. The integration of a pMUTIN vector into the target gene has three consequences: (1) the target gene is inactivated; (2) lacZ becomes transcriptionally fused to the gene, allowing its expression pattern to be monitored; (3) the Pspac promoter controls the transcription of downstream genes in an IPTG-dependent fashion. This last feature is important because B. subtilis genes are often organized in operons. The potential polar effects generated by the integration of the vectors can be alleviated by addition of IPTG. Also, conditional mutants of essential genes can be obtained by integrating pMUTIN vectors upstream of the target gene. The vectors are currently being used for systematic inactivation of genes without known function within the B. subtilis European consortium. pMUTIN characteristics and the inactivation of eight genes in the resA-serA region of the chromosome are presented. PMID- 9846746 TI - The product of the yvoC (gerF) gene of Bacillus subtilis is required for spore germination. AB - All known gerF mutations affecting Bacillus subtilis spore germination have been mapped, by a combination of recombination and complementation analysis, to yvoC (Igt), a gene belonging to the yvoB (ptsK) yvoC (Igt) yvoDEF operon. Examination of the properties of null mutants confirmed that the only gene in the operon that affects germination is yvoC, which encodes a homologue of known prelipoprotein diacylglyceryl transferases. As several germination proteins (GerAC, GerBC, GerKC, GerD) are predicted lipoproteins, it is not unreasonable to assume that a defect in prelipoprotein processing will affect spore germination. Two other null mutants in this chromosomal region showed a clear phenotype: the nagA gene is required for growth on N-acetylglucosamine, whereas a null mutation in yvoB (ptsK) affects colony formation from single cells. PMID- 9846747 TI - The kdgRKAT operon of Bacillus subtilis: detection of the transcript and regulation by the kdgR and ccpA genes. AB - Transcription of a new catabolic operon in Bacillus subtilis, involved in the late stages of galacturonic acid utilization, has been studied. The operon consists of four genes: kdgR, encoding the putative regulator protein; kdgK, encoding 2-keto-3-deoxygluconate kinase; kdgA, encoding 2-keto-3-deoxygluconate-6 phosphate aldolase; and kdgT, encoding a transporter. These four genes are organized in one transcriptional unit and map at 198 degrees of the B. subtilis chromosome. Primer extension experiments and Northern blot analysis show that an active sigmaA-dependent promoter precedes kdgR and transcription is terminated at the putative p-independent terminator downstream of kdgT. The operon is negatively regulated by the kdgR and ccpA gene products, which belong to the LacI family of transcription regulators. The expression of the genes in this operon can be induced by galacturonate and strongly repressed when glucose is present in the growth medium. Knockout mutations in genes kdgR and ccpA remove, respectively, the effects of galacturonate and glucose on the transcription of this operon. PMID- 9846748 TI - Siderotyping of fluorescent pseudomonads: characterization of pyoverdines of Pseudomonas fluorescens and Pseudomonas putida strains from Antarctica. AB - Five independent fluorescent pseudomonad isolates originating from Antarctica were analysed for their pyoverdine systems. A pyoverdine-related siderotyping, which involved pyoverdine-induced growth stimulation, pyoverdine-mediated iron uptake, pyoverdine analysis by electrophoresis and isoelectric focusing, revealed three different pyoverdine-related siderotypes among the five isolates. One siderotype, including Pseudomonas fluorescens 1W and P. fluorescens 10CW, was identical to that of P. fluorescens ATCC 13525. Two other strains, P. fluorescens 9AW and Pseudomonas putida 9BW, showed identical pyoverdine-related behaviour to each other, whereas the fifth strain, P. fluorescens 51W, had unique features compared to the other strains or to a set of 12 fluorescent Pseudomonas strains used as comparison material. Elucidation of the structure of the pyoverdines produced by the Antarctic strains supported the accuracy of the siderotyping methodology by confirming that pyoverdines from strains 1W and 10CW had the same structures as the P. fluorescens ATCC 13525 pyoverdine, whereas the 9AW and 9BW pyoverdines are probably identical with the pyoverdine of P. fluorescens strain 244. Pyoverdine from strain 51W appeared to be a novel pyoverdine since its structure was different from all previously established pyoverdine structures. Together with the conclusion that the Antarctic Pseudomonas strains have no special features at the level of their pyoverdines and pyoverdine-mediated iron metabolism compared to worldwide strains, the present work demonstrates that siderotyping provides a rapid means of screening for novel pyoverdines. PMID- 9846749 TI - PhhC is an essential aminotransferase for aromatic amino acid catabolism in Pseudomonas aeruginosa. AB - The phhC gene of Pseudomonas aeruginosa encodes a protein which is a member of the Family I aminotransferases. At high expression levels in the heterologous Escherichia coli system, PhhC can compensate for the absence of AspC (which functions in L-aspartate biosynthesis) and TyrB (which functions in aromatic biosynthesis). In the native organism, PhhC is essential for catabolism of either L-tyrosine or L-phenylalanine, as demonstrated by gene inactivation. This catabolic function of PhhC is consistent with its inclusion as the distal gene in the inducible phenylalanine hydroxylase operon. The presence of PhhC for catabolism of aromatic amino acids is required in spite of an existing multiplicity of other P. aeruginosa aminotransferases having a similar pattern of broad substrate specificity in vitro. This implies a spatial orientation of PhhC that effectively specializes it for aromatic amino acid catabolism. PMID- 9846750 TI - Dihydroaeruginoic acid synthetase and pyochelin synthetase, products of the pchEF genes, are induced by extracellular pyochelin in Pseudomonas aeruginosa. AB - The siderophore pyochelin of Pseudomonas aeruginosa is derived from one molecule of salicylate and two molecules of cysteine. Two cotranscribed genes, pchEF, encoding peptide synthetases have been identified and characterized. pchE was required for the conversion of salicylate to dihydroaeruginoate (Dha), the condensation product of salicylate and one cysteine residue and pchF was essential for the synthesis of pyochelin from Dha. The deduced PchE (156 kDa) and PchF (197 kDa) proteins had adenylation, thiolation and condensation/cyclization motifs arranged as modules which are typical of those peptide synthetases forming thiazoline rings. The pchEF genes were coregulated with the pchDCBA operon, which provides enzymes for the synthesis (PchBA) and activation (PchD) of salicylate as well as a putative thioesterase (PchC). Expression of a translational pchE'-'lacZ fusion was strictly dependent on the PchR regulator and was induced by extracellular pyochelin, the end product of the pathway. Iron replete conditions led to Fur (ferric uptake regulator)-dependent repression of the pchE'-'lacZ fusion. A translational pchD'-'lacZ fusion was also positively regulated by PchR and pyochelin and repressed by Fur and iron. Thus, autoinduction by pyochelin (or ferric pyochelin) and repression by iron ensure a sensitive control of the pyochelin pathway in P. aeruginosa. PMID- 9846751 TI - Molecular and functional analysis of pTAV320, a repABC-type replicon of the Paracoccus versutus composite plasmid pTAV1. AB - The second replicator region of the native plasmid pTAV1 of Paracoccus versutus has been identified thus proving the composite nature of this replicon. The minimal replicon designated pTAV320 (4.3 kb) was cloned and sequenced. pTAV320 encodes three putative proteins--RepA, RepB and RepC. This replicator region shows strong structural and functional similarity to repABC-type replicons found in several Agrobacterium and Rhizobium plasmids. The origin of replication appears to be localized within the coding sequence of the repC gene. RepC was shown to be essential for replication. RepA and RepB were necessary for stable maintenance of the plasmid, which implies a role in active partitioning. The presence of the complete sequence of pTAV320 (in its non-replicative form) could stabilize in cis pTAV202, a mini-replicon derived from the other pTAV1 replicator region. Deletions introduced into the repC gene abolished the 'stabilizing' activity of pTAV320, suggesting that the centromere-like sequence, necessary for partitioning, might be localized within this gene. The two replicator regions of pTAV1 (pTAV320 and pTAV202) expressed incompatibility towards the parental plasmid but were compatible in trans in P. versutus cells. The pTAV320 replicon can be maintained in several Paracoccus, Agrobacterium, Rhizobium and Rhodobacter strains in addition to P. versutus. PMID- 9846752 TI - Isolation and characterization of Staphylococcus aureus starvation-induced, stationary-phase mutants defective in survival or recovery. AB - Ten Staphylococcus aureus mutants, defective in the starvation-induced stationary phase of growth were isolated from two independent Tn917-LTV1 transposon insertion libraries and were designated suv as they had apparent survival defects. Seven of these mutants were defective under amino-acid-limiting conditions alone. Two mutants (suv-3 and suv-20) demonstrated lower plating efficiency when starved for glucose, phosphate or amino acids and one mutant (suv 11) had reduced plating efficiency after amino acid or glucose starvation. All of the mutants tested were as resistant to hydrogen peroxide assault as the parent, but six were more sensitive to low pH conditions. All the mutants were physically mapped on the S. aureus chromosome using PFGE. Chromosomal DNA flanking the Tn917 LTV1 insertion sites was rescued by cloning into Escherichia coli. DNA sequence analysis resulted in the identification of a number of transposon-disrupted ORFs encoding putative components such as superoxide dismutase (suv-1), haem A synthase (suv-3), a component of the SOS response (suv-9) and hypoxanthine guanine phosphoribosyltransferase (suv-20). The Tn917-LTV1 insertion created lacZ transcriptional fusions for some of the stationary-phase loci. Expression analysis indicated that suv-4 was induced at mid-exponential phase, whereas suv-3 and suv-11 were induced at the onset of stationary phase. The possible roles of these suv components in stationary-phase survival or recovery is discussed. PMID- 9846753 TI - A 1,3-beta-glucanosyltransferase isolated from the cell wall of Aspergillus fumigatus is a homologue of the yeast Bgl2p. AB - A 1,3-beta-glucanosyltransferase which introduces intrachain 1,6-beta linkages into 1,3-beta-glucan was isolated from the cell wall of Aspergillus fumigatus. The biochemical and molecular characterization of the A. fumigatus transferase showed it was homologous to the Saccharomyces cerevisiae and Candida albicans transferase Bgl2p. A null mutant constructed in A. fumigatus by gene replacement did not show a distinct phenotype from the parental strain. The putative function of this major cell-wall-associated protein is discussed. PMID- 9846754 TI - Molecular constituents of the replication apparatus in the plasmodium of Physarum polycephalum: identification by photoaffinity labelling. AB - The plasmodium of Physarum polycephalum has long been considered a model system for syncytically growing cells, but important details of the DNA replication apparatus, such as the DNA polymerase epsilon and other replication factors, have not been detected. In this study, a new variation of photoaffinity labelling and immunoblotting was used to detect DNA polymerases and other factors in nuclear extracts of P. polycaphalum. Proteins were specifically cross-linked with photoreactive arylazido-dCMP residues incorporated during extension of template primer DNA. The DNA synthesized in situ was 32P-labelled. After nucleolytic removal of protruding DNA, the proteins were separated by SDS-gel electrophoresis, electroblotted on membranes and subjected to autoradiography. The alpha, delta, epsilon and beta-like DNA polymerases were labelled, as were histones and replication-factor-like proteins. Cytoplasmic extracts were devoid of these species. Abundant proliferating-cell nuclear antigen and replication protein A large subunit were labelled and found to be of unusual mass. A number of subunits of purified DNA polymerase holoenzymes were labelled. In contrast, only the DNA-polymerizing subunits could be labelled in nuclear extracts. Higher order complexes in the nuclear extract may make subunits inaccessible to photo cross-linking. Complex formation is promoted by beta-poly(L-malate), a plasmodium specific putative storage and carrier molecule that supports DNA replication in the synchronized nuclei. Percoll, a polyvinylpyrrolidone-coated colloidal silica, partially disrupted these complexes. A 200 kDa fragment of DNA polymerase epsilon and a 135 kDa beta-like DNA polymerase did not participate in the complexes, suggesting functions unlike those of the other polymerases. DNA polymerase molecules were intact during proliferation of plasmodia, but were nicked before their clearance from the nuclei at growth arrest. PMID- 9846755 TI - A Mycobacterium tuberculosis operon encoding ESAT-6 and a novel low-molecular mass culture filtrate protein (CFP-10). AB - The early secreted antigenic target 6 kDa protein (ESAT-6) is a potent T-cell protein antigen synthesized by Mycobacterium tuberculosis. Its corresponding gene (esat-6) is located in RD1, a 10 kb DNA region deleted in the attenuated tuberculosis vaccine strain Mycobacterium bovis BCG. The promoter region of M. tuberculosis esat-6 was cloned and characterized. A new gene, designated lhp and cotranscribed with esat-6, was identified. Moreover, computer searches in the M. tuberculosis genome identified 13 genes related to the lhp/esat-6 operon, defining a novel gene family. The transcription initiation sites of the lhp/esat 6 operon were mapped using M. tuberculosis RNA. The corresponding promoter signals were not recognized in Mycobacterium smegmatis, in which transcription of lhp/esat-6 is initiated at different locations. The M. tuberculosis lhp gene product was identified as CFP-10, a low-molecular-mass protein found in the short term culture filtrate. These results show that the genes encoding CFP-10 and ESAT 6 are transcribed together in M. tuberculosis and that both code for small exported proteins. PMID- 9846756 TI - Isolation of carbon- and nitrogen-deprivation-induced loci of Sinorhizobium meliloti 1021 by Tn5-luxAB mutagenesis. AB - Soil bacteria, such as Sinorhizobium meliloti, are subject to variation in environmental conditions, including carbon- and nitrogen-deprivation. The ability of bacteria to sense changes in their environment and respond accordingly is of vital importance to their survival and persistence in the soil and rhizosphere. A derivative of Tn5 which creates transcriptional fusions to the promoterless luxAB genes was used to mutagenize S. meliloti 1021 and 5000 insertion mutants were subsequently screened for gene fusions induced by selected environmental stresses. The isolation of 21 gene fusions induced by nitrogen-deprivation and 12 induced by carbon-deprivation is described. Cloning and partial DNA sequence analysis of the transposon-tagged loci revealed a variety of novel genes, as well as S. meliloti genes with significant similarity to known bacterial loci. In addition, nodule occupancy studies were carried out with selected Tn5-luxAB insertion mutants to examine the role of the tagged genes in competition. PMID- 9846757 TI - Functional characterization of the Erwinia chrysanthemi OutS protein, an element of a type II secretion system. AB - Secretion of pectate lyases and a cellulase occurs in Erwinia chrysanthemi through a type II secretion machinery, the Out system. Proper insertion of the secretin OutD in the outer membrane requires the presence of OutS. OutS is an outer-membrane lipoprotein that interacts directly with OutD. Using ligand blotting experiments, it has been shown that this interaction requires at least the 62 C-terminal amino acids of OutD. When this domain was added to the C terminal extremity of the secreted pectate lyase PelD, the construct was stabilized by OutS but not inserted into the outer membrane. Thus, this domain is sufficient to interact with OutS but it is unable to confer the ability to be inserted into the outer membrane in the presence of OutS. A screen for outS mutants unable to secrete pectate lyases gave only mutants unable to properly localize OutD in the outer membrane and no mutant in the protection function. Thus, the interaction between OutS and OutD can probably not be abolished by the mutation of a single amino acid, and the insertion of OutD in the outer membrane may require additional proteins. PMID- 9846759 TI - Global perspective from International AIDS Society president Mark Wainberg. Interview by Dennis Blakesley. PMID- 9846760 TI - Reports focus on bad, good in children's health. PMID- 9846758 TI - Malate synthase from Streptomyces clavuligerus NRRL3585: cloning, molecular characterization and its control by acetate. AB - Malate synthase is a key enzyme of the glyoxylate cycle, which is an anaplerotic pathway essential for growth on acetate as the sole carbon source. The aceB gene, encoding malate synthase from Streptomyces clavuligerus NRRL 3585, was cloned using PCR and fully sequenced. The ORF obtained encodes 541 amino acids with a deduced Mr of 60000, consistent with the observed Mr (62000-64000) of most malate synthase enzymes reported so far. The aceB gene has a high G+C content (71.5 mol%), especially in the third codon position. A 50 bp region upstream of the malate synthase ORF was predicted to be a prokaryotic promoter region. The relationship between carbon source, antibiotic (cephalosporin) biosynthesis and malate synthase activity was investigated. Growth of S. clavuligerus on acetate as the major carbon source was delayed, compared to that on glycerol. Furthermore, high levels of malate synthase activity were associated with the presence of acetate in the growth medium. Growth on acetate also resulted in lower levels of cephalosporin production, compared to that on glycerol. The cloned S. clavuligerus aceB gene was expressed in Escherichia coli BL21(DE3). Transformants exhibited an approximately 71-fold increase in malate synthase activity, compared to the control, thereby demonstrating high-level expression of soluble and enzymically active malate synthase in the heterologous host. PMID- 9846761 TI - New solution for preserving red blood cells. PMID- 9846762 TI - From the Food and Drug Administration. PMID- 9846763 TI - From the Centers for Disease Control and Prevention. Influenza and pneumococcal vaccination levels among adults aged > or = 65 years--United States, 1997. PMID- 9846764 TI - From the Centers for Disease Control and Prevention. Trends in sexual risk behaviors among high school students--United States, 1991-1997. PMID- 9846765 TI - Primary prevention of postmenopausal osteoporosis. PMID- 9846766 TI - Medical practices disappearing in the South. PMID- 9846767 TI - Free PSA for detecting prostate cancer. PMID- 9846768 TI - Free PSA for detecting prostate cancer. PMID- 9846769 TI - Free PSA for detecting prostate cancer. PMID- 9846770 TI - Permission and confidentiality in publishing pedigrees. PMID- 9846772 TI - Collective bargaining for residents. PMID- 9846773 TI - Risk of invasive streptococcal disease. PMID- 9846774 TI - Plasma D-dimer in the diagnosis of deep vein thrombosis. PMID- 9846775 TI - Adult purpura fulminans and digital necrosis associated with sepsis and the factor V mutation. PMID- 9846776 TI - An HIV-resistant allele is exceptionally frequent in New Guinean highlanders. PMID- 9846777 TI - Gabapentin for the symptomatic treatment of painful neuropathy in patients with diabetes mellitus: a randomized controlled trial. AB - CONTEXT: Pain is the most disturbing symptom of diabetic peripheral neuropathy. As many as 45% of patients with diabetes mellitus develop peripheral neuropathies. OBJECTIVE: To evaluate the effect of gabapentin monotherapy on pain associated with diabetic peripheral neuropathy. DESIGN: Randomized, double-blind, placebo-controlled, 8-week trial conducted between July 1996 and March 1997. SETTING: Outpatient clinics at 20 sites. PATIENTS: The 165 patients enrolled had a 1- to 5-year history of pain attributed to diabetic neuropathy and a minimum 40 mm pain score on the Short-Form McGill Pain Questionnaire visual analogue scale. INTERVENTION: Gabapentin (titrated from 900 to 3600 mg/d or maximum tolerated dosage) or placebo. MAIN OUTCOME MEASURES: The primary efficacy measure was daily pain severity as measured on an 11-point Likert scale (0, no pain; 10, worst possible pain). Secondary measures included sleep interference scores, the Short Form McGill Pain Questionnaire scores, Patient Global Impression of Change and Clinical Global Impression of Change, the Short Form-36 Quality of Life Questionnaire scores, and the Profile of Mood States results. RESULTS: Eighty four patients received gabapentin and 70 (83%) completed the study; 81 received placebo and 65 (80%) completed the study. By intent-to-treat analysis, gabapentin treated patients' mean daily pain score at the study end point (baseline, 6.4; end point, 3.9; n = 82) was significantly lower (P<.001) compared with the placebo-treated patients' end-point score (baseline, 6.5; end point, 5.1; n = 80). All secondary outcome measures of pain were significantly better in the gabapentin group than in the placebo group. Additional statistically significant differences favoring gabapentin treatment were observed in measures of quality of life (Short Form-36 Quality of Life Questionnaire and Profile of Mood States). Adverse events experienced significantly more frequently in the gabapentin group were dizziness (20 [24%] in the gabapentin group vs 4 [4.9%] in the control group; P<.001) and somnolence (19 [23%] in the gabapentin group vs 5 [6%] in the control group; P = .003). Confusion was also more frequent in the gabapentin group (7 [8%] vs 1 [1.2%]; P = .06). CONCLUSION: Gabapentin monotherapy appears to be efficacious for the treatment of pain and sleep interference associated with diabetic peripheral neuropathy and exhibits positive effects on mood and quality of life. PMID- 9846778 TI - Gabapentin for the treatment of postherpetic neuralgia: a randomized controlled trial. AB - CONTEXT: Postherpetic neuralgia (PHN) is a syndrome of often intractable neuropathic pain following herpes zoster (shingles) that eludes effective treatment in many patients. OBJECTIVE: To determine the efficacy and safety of the anticonvulsant drug gabapentin in reducing PHN pain. DESIGN: Multicenter, randomized, double-blind, placebo-controlled, parallel design, 8-week trial conducted from August 1996 through July 1997. SETTING: Sixteen US outpatient clinical centers. PARTICIPANTS: A total of 229 subjects were randomized. INTERVENTION: A 4-week titration period to a maximum dosage of 3600 mg/d of gabapentin or matching placebo. Treatment was maintained for another 4 weeks at the maximum tolerated dose. Concomitant tricyclic antidepressants and/or narcotics were continued if therapy was stabilized prior to study entry and remained constant throughout the study. MAIN OUTCOME MEASURES: The primary efficacy measure was change in the average daily pain score based on an 11-point Likert scale (0, no pain; 10, worst possible pain) from baseline week to the final week of therapy. Secondary measures included average daily sleep scores, Short-Form McGill Pain Questionnaire (SF-MPQ), Subject Global Impression of Change and investigator-rated Clinical Global Impression of Change, Short Form-36 (SF-36) Quality of Life Questionnaire, and Profile of Mood States (POMS). Safety measures included the frequency and severity of adverse events. RESULTS: One hundred thirteen patients received gabapentin, and 89 (78.8%) completed the study; 116 received placebo, and 95 (81.9%) completed the study. By intent-to treat analysis, subjects receiving gabapentin had a statistically significant reduction in average daily pain score from 6.3 to 4.2 points compared with a change from 6.5 to 6.0 points in subjects randomized to receive placebo (P<.001). Secondary measures of pain as well as changes in pain and sleep interference showed improvement with gabapentin (P<.001). Many measures within the SF-36 and POMS also significantly favored gabapentin (P< or =.01). Somnolence, dizziness, ataxia, peripheral edema, and infection were all more frequent in the gabapentin group, but withdrawals were comparable in the 2 groups (15 [13.3%] in the gabapentin group vs 11 [9.5%] in the placebo group). CONCLUSIONS: Gabapentin is effective in the treatment of pain and sleep interference associated with PHN. Mood and quality of life also improve with gabapentin therapy. PMID- 9846779 TI - Abdominal adiposity and coronary heart disease in women. AB - CONTEXT: Obesity is a well-established risk factor for coronary heart disease (CHD), but whether regional fat distribution contributes independently to risk remains unclear. OBJECTIVE: To compare waist-hip ratio (WHR) and waist circumference in determining risk of CHD in women. DESIGN AND SETTING: Prospective cohort study among US female registered nurses participating in the Nurses' Health Study conducted between 1986, when the nurses completed a questionnaire, and follow-up in June 1994. PARTICIPANTS: A total of 44702 women aged 40 to 65 years who provided waist and hip circumferences and were free of prior CHD, stroke, or cancer in 1986. MAIN OUTCOME MEASURES: Incidence of CHD (nonfatal myocardial infarction or CHD death). RESULTS: During 8 years of follow up 320 CHD events (251 myocardial infarctions and 69 CHD deaths) were documented. Higher WHR and greater waist circumference were independently associated with a significantly increased age-adjusted risk of CHD. After adjusting for body mass index (BMI) (defined as weight in kilograms divided by the square of height in meters) and other cardiac risk factors, women with a WHR of 0.88 or higher had a relative risk (RR) of 3.25 (95% confidence interval [CI], 1.78-5.95) for CHD compared with women with a WHR of less than 0.72. A waist circumference of 96.5 cm (38 in) or more was associated with an RR of 3.06 (95% CI, 1.54-6.10). The WHR and waist circumference were independently strongly associated with increased risk of CHD also among women with a BMI of 25 kg/m2 or less. After adjustment for reported hypertension, diabetes, and high cholesterol level, a WHR of 0.76 or higher or waist circumference of 76.2 cm (30 in) or more was associated with more than a 2-fold higher risk of CHD. CONCLUSIONS: The WHR and waist circumference are independently associated with risk of CHD in women. PMID- 9846780 TI - Secular trends in preterm birth: a hospital-based cohort study. AB - CONTEXT: Canada and the United States have reported a recent increase in the incidence of preterm birth, but the reasons for this increase are unknown. OBJECTIVE: To assess secular trends in preterm birth and its potential determinants. DESIGN: Hospital-based cohort study. SETTING: Canadian tertiary care university teaching hospital, 1978-1996. PARTICIPANTS: A total of 65574 nonreferred live births and stillbirths. MAIN OUTCOME MEASURES: Changes in occurrence of preterm birth, before and after adjustment for changes in method of gestational age assessment, obstetric intervention, registration of births weighing less than 500 g, and sociodemographic, behavioral, and clinical determinants. RESULTS: A crude secular increase in preterm births was seen for births less than 37, 34, and 32 completed weeks using 3 alternative gestational age estimation methods. Based on an algorithm incorporating both menstrual and early ultrasound gestational age estimates, rates increased from 6.6% to 9.8% for births at less than 37 weeks' gestation, 1.7% to 2.3% at less than 34 weeks, and 1.0% to 1.2% at less than 32 weeks. Exclusion of births weighing less than 500 g and those with induction or preterm cesarean delivery without labor before each of the corresponding gestational age cutoffs eliminated the secular trends for births before 34 and 32 weeks and attenuated the trend for births before 37 weeks. Nearly half of the remaining trend for births before 37 weeks was accounted for by the increasing use of early ultrasound dating. The residual trend was eliminated after controlling for secular increases in unmarried status and the proportion of women aged 35 years or older. These factors, combined with a decrease in alcohol consumption and increases in histological chorioamnionitis and cocaine use, appear to have counteracted a reduction in preterm birth since the mid-1980s that otherwise would have been observed. CONCLUSIONS: This hospital's increase in preterm births since 1978 parallels increases reported in population-based national studies from the United States and Canada. This trend appears largely attributable to the increasing use of early ultrasound dating, preterm induction and preterm cesarean delivery without labor, and changes in sociodemographic and behavioral factors. PMID- 9846781 TI - Sexual abuse of boys: definition, prevalence, correlates, sequelae, and management. AB - OBJECTIVE: To clarify the definition of sexual abuse of boys, update estimates of its prevalence, and explore critically its reported correlates, sequelae, and management. DATA SOURCES: Studies from 1985 to 1997 were identified using OVID MEDLINE and OVID-CINAHL. The search terms used were sexual abuse, sexual victimization, and sexual assault. Constraints applied included English language, human male subjects, age younger than 19 years, and North American samples. STUDY SELECTION: Publications were included for review if they appeared in peer reviewed journals; had clear research designs; reported results for at least 20 male subjects; and were not reviews, perspectives, theoretical treatises, editorials, or letters. DATA EXTRACTION: Study types and sampling methods were categorized using well-established definitions. Preference was given to studies with large samples, with case-control or cohort designs, and/or with adjustment for effect modifiers or confounders. DATA SYNTHESIS: We identified 166 studies representing 149 sexual abuse samples. Studies were methodologically limited and definitions of sexual abuse varied widely. Prevalence estimates varied widely (by definition used and population studied), ranging from 4% to 76%. Boys at highest risk were younger than 13 years, nonwhite, of low socioeconomic status, and not living with their fathers. Perpetrators tended to be known but unrelated males. Abuse frequently occurred outside the home, involved penetration, and occurred more than once. Sequelae included psychological distress, substance abuse, and sexually related problems. Evaluation of management strategies was limited. CONCLUSIONS: Sexual abuse of boys appears to be common, underreported, underrecognized, and undertreated. Future study requires clearer definitions of abuse, improved sampling, more rigorous data collection, more sophisticated data analyses, and better assessment of management and treatment strategies. Regardless, health care professionals should be more aware of and sensitive to the possibility of sexual abuse in their male patients. PMID- 9846782 TI - Symptomatic treatment of painful neuropathy. PMID- 9846783 TI - Improving research, policy, and practice to understand child sexual abuse. PMID- 9846785 TI - Selma Deitch, MD, MPH: children first. PMID- 9846786 TI - On the brink: the costs of medical education. PMID- 9846787 TI - The increasing debt of medical students: how much is too much? PMID- 9846788 TI - Into the deep well: the evolution of medical school loan debt. PMID- 9846789 TI - The medical resident workload: the case of New York State. PMID- 9846790 TI - JAMA patient page: sexual abuse. PMID- 9846791 TI - 6-month use of 0.2% delmopinol hydrochloride in comparison with 0.2% chlorhexidine digluconate and placebo (II). Effect on plaque and salivary microflora. AB - This double-blind, randomised, 6-month clinical trial with parallel group design in 68 subjects with gingivitis was conducted to study the effects on the oral flora of delmopinol hydrochloride 2 mg/ml (0.2% w/v, Decapinol Mouthwash), when used for partly supervised mouthrinsing in comparison with placebo and chlorhexidine digluconate 2 mg/ml (0.2% w/v, Hibitane Dental, ICI Pharmaceuticals, UK). Apart from estimating the total cultivable microbial dental plaque flora and salivary flora, analyses were focused on bacterial groups associated with gingivitis/periodontitis and dental caries. Furthermore, the presence of staphylococci, gram-negative enteric bacteria and yeasts in saliva were evaluated. The minimal inhibitory concentration (MIC) was determined for isolates belonging to the predominating micro-organisms in samples of both dental plaque and saliva. In relation to the findings in the placebo group, the use of delmopinol during the rinsing period did not produce an undesirable shift in the bacterial populations considered to be related to dental caries or periodontal diseases. These groups remained virtually unchanged during the study. In relation to the observations in the placebo group, slight reductions in the total cultivable plaque and salivary flora were observed during the study and no change was found in the ratio total anaerobically/aerobically cultivable microbial flora. Furthermore, no increased growth in staphylococci, enteric bacteria or yeasts was observed in the saliva samples. The pattern of changes taking place in the composition of the plaque and salivary microbial flora in samples from the participants rinsing with chlorhexidine were in most aspects similar to that observed in the delmopinol group. In the delmopinol group, no microbiologically significant changes were observed over time in the MIC-values for the isolates, neither in the plaque nor in the saliva samples, which indicates that no adaptation to delmopinol had taken place during the rinsing period. Similar observations were made for the plaque isolates in samples from the participants in the chlorhexidine group. On the other hand, when gram-positive and catalase negative cocci from the saliva samples of the latter group were tested against chlorhexidine, 4-6 times higher MIC-values were obtained at 3 and 6 months both when compared to baseline and in comparison with the other two rinsing groups (p<0.01 or p<0.05). Neither delmopinol nor chlorhexidine showed any residual effect on the studied microbial groups in the plaque and the saliva samples 3 months after the end of treatment. In conclusion, delmopinol was accompanied by a composition of the plaque and salivary flora associated with healthy conditions in the oral cavity. PMID- 9846792 TI - Reliability of linear alveolar bone loss measurements of mandibular posterior teeth from digitized bitewing radiographs. AB - Observer reliability in performing linear measurements between the cementoenamel junction and alveolar crest was determined for mandibular posterior teeth from digitized clinical bitewing radiographs acquired during recall examinations. 6 measurements (corresponding to traditional probing measurements) were made per tooth by 3 observers. Mesial and distal measurements made to the most coronal aspects of the alveolar crest were the most reliable and least biased. As was anticipated, intra-observer reliability was better than inter-observer reliability although the 3 observers of our study were able to detect a significant mean change (0.1 mm, p<0.0001) in alveolar bone height over a 1-year period for 10 patients. For our most reliable and unbiased measurements (mesial measurements to the alveolar crest), a change of 0.54 mm (90th percentile) would be required to indicate change at a site from one time to the next. Based on the reliability of our digital radiographic measurements, with the alpha error rate set at 0.05 and beta at 0.20, a difference in alveolar bone height of 0.3 mm could be detected with a patient sample size of between 13 (best case) and 54 (worst case). PMID- 9846793 TI - Additional clinical and microbiological effects of amoxicillin and metronidazole after initial periodontal therapy. AB - The aims of this study were to evaluate the clinical and microbiological effects of initial periodontal therapy (IT) and to determine the additional effects of systemic amoxicillin (Flemoxin Solutab) 375 mg TID plus metronidazole 250 mg TID therapy, in patients with adult Actinobacillus actinomycetemcomitans (Aa) associated periodontitis in conjunction with either Porphyromonas gingivalis (Pg), Bacteroides forsythus (Bf) and/or Prevotella intermedia (Pi). In addition the adverse effects of the antimicrobial therapy were also documented. A total of 22 patients were enrolled. The deepest, bleeding pocket in each quadrant was selected and at these 4 experimental sites clinical measurements and microbiological testing was carried out at baseline, after (IT), i.e., 21 weeks after baseline, and after antimicrobial therapy (AM), i.e., 35 weeks after baseline. At baseline, the mean plaque index (PI) amounted 0.5, 0.1 after IT and 0.3 after systemic AM. The mean bleeding index decreased from 1.6 to 1.2 after IT and a further decrease to 0.7 after AM was noted. Suppuration was completely eliminated after AM. The mean change of probing pocket depth (PPD) after IT amounted 1.4 mm and was further reduced with an additional mean change of 1.1 mm after medication. Clinical attachment gain was 1.1 mm after IT and an additional 0.9 mm was observed after AM. One of the 22 Aa positive patients and 4 of 17 Pg positive patients became negative for these species after IT. The number of patients with detectable Pi decreased from 16 to 10 after IT. After AM, in comparison to baseline, suppression below detection level for Aa was achieved in 19 out of 22, for Pg in 9 out of 17, for Bf in 13 out of 14, and for Pi in 11 out of 16 patients. By contrast, higher frequencies of Peptostreptococcus micros and Fusobacterium nucleatum were found after AM. On the basis of the microbiological results the study group was separated into 2 subgroups: group A consisted of subjects who had no detectable levels of Aa, Pg, Bf and <5% of Pi after AM. Group B consisted of those who still showed presence of one of these 3 species and/or > or =5% levels of Pi. After AM, group B had significantly higher PI, BI, PPD and CAL scores then group A. It is concluded that group A showed low plaque scores and no detectable periodontal pathogens. This microbiological condition has been associated with a long-term stable periodontium. PMID- 9846795 TI - The description of a unique population with a very high prevalence of localized juvenile periodontitis. AB - The reported prevalence of localized juvenile periodontitis (LJP) amongst teenagers and young adults varies greatly. The etiology of LJP has been related to Actinobacillus actinomycetemcomitans (Aa), and it has also been suggested that there may be a transmission of Aa within families resulting in the familial distribution of the disease. This study describes the high prevalence of LJP in adolescents, 12-20 years of age, from a group of nuclear families living and functioning in a closed, closely knit community. The survey was carried out on a population of teenagers that had attended the same school and their siblings. All students attending that school and their siblings were examined. They were given a periodontal examination and a questionnaire relating to their demographic details and their personal oral hygiene habits. The periodontal examination was limited to the incisors and first molar teeth. Plaque index (PlI), gingival index (GI), the presence or absence of bleeding on probing (BOP), probing pocket depth (PPD) and recession were measured. All patients having at least two of the examined sites with probing pocket depth > or =5 mm or one site > or =6 mm were considered as possible sufferers from LJP and had a full mouth periapical radiographic survey carried out using a paralleling technique to confirm the diagnosis. At the sites with probing pocket depth > or =5 mm, a Shei ruler was used to measure the % of the root coronal to the alveolar bone. A cut off point of > or =20% was used as a measure of true bone loss confirming the clinical diagnosis of LJP. 86 individuals from 30 families comprised the population of interest. There were 44 males and 42 females with a mean age of 14.7+/-2.3. Of the 86 individuals examined, 33 individuals from 15 families were diagnosed as having LJP (38.4%). None of the individuals examined showed any evidence of the generalized form of juvenile periodontitis. The mean age of the LJP patients was 15+/-2.3 yrs. with a 1:1.75 male to female ratio. Except for 2 pairs of families with genetic ties, no familial connections could be traced between the different nuclear families affected by LJP despite repeated and intensive questioning. There were no significant differences in the PlI and the GI between the groups while the LJP group had significantly higher BOP, PPD and PAL than the non-LJP group. These finding strongly suggest an environmental influence in the etiology of the disease. PMID- 9846794 TI - Relationship between C-telopeptide pyridinoline cross-links (ICTP) and putative periodontal pathogens in periodontitis. AB - Crevicular fluid pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) is predictive for future alveolar bone loss in experimental periodontitis in dogs. The present study sought to relate ICTP to a panel of subgingival species in subjects exhibiting various clinical presentations such as health (n=7), gingivitis (n=8) and periodontitis (n=21). 28 subgingival plaque and GCF samples were taken from mesiobuccal sites in each of 36 subjects. The presence and levels of 40 subgingival taxa were determined in plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. GCF ICTP levels were quantified using radioimmunoassay (RIA). Clinical assessments made at the same sites included: BOP, gingival redness, plaque, pocket depth, and attachment level. Differences among ICTP levels in the 3 subject groups were sought using the Kruskal-Wallis test. Relationships between ICTP levels and clinical parameters as well as subgingival species were determined by regression analysis. The results demonstrated significant differences among disease categories for GCF ICTP levels for healthy (1.1+0.6 pg/site (mean+/-SEM)) gingivitis (14.8+/-6.6 pg/site) and periodontitis subjects (30.3+5.7 pg/site) (p= 0.0017). ICTP levels related modestly to several clinical parameters. Regression analysis indicated that ICTP levels correlated strongly with mean subject levels of several periodontal pathogens including B. forsythus, P. gingivalis, P. intermedia, P. nigrescens and T. denticola (p<0.01). The data indicate that there is a positive relationship between the putative bone resorptive marker ICTP and periodontal pathogens. PMID- 9846796 TI - The inhibitory effect of combining chlorhexidine and hydrogen peroxide on 3-day plaque accumulation. AB - In a blind, randomised, 4-cell, cross-over study, the effect of rinsing with a perborate solution (1.9 g sodium perborate-monohydrate dissolved in 30 ml water/Bocasan, Oral-B) on the in vivo plaque-inhibiting effect of 0.12% chlorhexidine (Oral-B) was examined. After a thorough professional prophylaxis including interdental cleaning, 12 subjects started to rinse according to 4 different regimens: regimen (C-P-C): chlorhexidine in the morning, perborate at noon and chlorhexidine in the evening; regimen (CP-CP): chlorhexidine immediately followed by perborate in the morning and in the evening; regimen (PC-PC): perborate immediately followed by chlorhexidine in the morning and in the evening; regimen (C-C): chlorhexidine in the morning and in the evening. No further oral hygiene measures were allowed for the next 72 h. After 72 h, the subjects were scored for plaque, and a washout period of 4 days followed; cross over was randomly assigned according to a Latin square design. Following this procedure, all subjects went through all 4 regimens. The regimens C-P-C and PC-PC resulted in significantly lower plaque-scores, 0.27 and 0.28 respectively, than regimen C-C (0.40). For the regimen CP-CP, the plaque-score was 0.28, which was not significantly different from the C-C regimen. The results suggest a positive interaction between chlorhexidine and hydrogen peroxide. Rinsing with a combination of 0.12% chlorhexidine (Oral-B) and a perborate solution (Bocasan Oral-B) can result in more effective short-term plaque growth inhibition than rinsing with chlorhexidine alone. PMID- 9846797 TI - The effects of Pain-Free Desensitizer on dentine permeability and tubule occlusion over time, in vitro. AB - The purpose of this in vitro study was to evaluate the efficacy of a new resin emulsion (Pain-Free Desensitizer) treatment for dentine hypersensitivity, for its ability to decrease dentine permeability. Crown segments were prepared from extracted, unerupted human 3rd molars by horizontal sectioning to remove occlusal enamel and the roots. The specimens were allocated in one of two groups: In group 1, the dentine surface was acid-etched to simulate the patent tubules of hypersensitive dentine. In group 2, the mineralized dentine surface was polished free of smear layer using a hydroxyapatite paste and ultrasonication. The hydraulic conductance of each specimen was then measured to obtain a pretreatment of control value. After a single treatment with resin desensitizer, the permeability was remeasured at 5 min, 1 day, 1 week and 1 month. Between measurements, the specimens were stored in buffer solution to simulate the solubilizing effects of saliva. Parallel specimens were followed by SEM examination. The results showed that a single treatment with resin desensitizer produced large, immediate, reductions in dentine permeability in both acid-etched and mineralized surfaces. In the acid-etched (group 1) specimens, the permeability returned to control values within 7 days, while the permeability of the group 2 specimens remained low even after 30 days of soaking. This simple treatment for occluding dentinal tubules may provide sufficient temporary reduction in dentine permeability to permit the development of natural desensitization. PMID- 9846798 TI - A dental implant: aluminium trioxide exhibited no effect on mouse reproductive and mutanogenic potential. AB - Several diseases as well as trauma can affect the composition and integrity of periodontal tissues leading eventually to the destruction of connective tissue matrix and cells, loss of attachment and resorption of alveolar bone, often followed by tooth loss. Replacement of the missing tooth could then be provided by endosseous dental implants healing in a form of osseo- or fibrosteal integration to the alveolar bone. Aluminium oxide ceramics, a form of endosseous implant, allows osseointegration type of healing and has demonstrated excellent biocompatibility. However, potential aluminium toxicity has been implicated in the pathogenesis of a number of clinical disorders and for this reason we examined the reproductive and mutagenic effect of aluminium trioxide ceramic implant in experimental mice. 720 female and 45 fertile male BALB-cAn NCR mice were included in the study. 3 experimental groups of fertile male mice (15 for each group) were treated with an intraperitoneal injection of aluminium trioxide (1 g/ kg of body weight, group I), with ethyl-methane-sulphonate as a positive control (200 mg/kg, group II) and with Tween-80 (10 mg/kg as a negative control, Group III). Each of the labeled male mice fertilized previously uncoupled female mice during 8 weeks (a pair per week) to facilitate appropriate pre- and post meiotic conditions of spermatogenesis to occur. Female mice were sacrificed with cervical dislocation at day 13 after fertilization. Immediately upon sacrifice the uterus was removed and the number of alive and healthy, or alive but mutated and/or dead embryos was computed to determine the dominant lethal or mutagenic effect. Animals treated with aluminium trioxide demonstrated similar effects on the reproductive and mutagenic capacity as the negative control, whereas the animals treated as positive controls exhibited significantly reduced reproductive and mutagenic capacity. Collectively, we concluded that aluminium trioxide has a very low rate of embryonal mortality and mutagenicity in mice. This finding is in general agreement with the biocompatibility of aluminium trioxide as an implant material. PMID- 9846799 TI - Clinical and microbiological findings in elderly subjects with gingivitis or periodontitis. AB - The objective of the present study was to study the supra- and subgingival microflora by culture and cDNA probe methods in 20 elderly subjects who were between 62 and 93 years of age. 10 of them had gingivitis only, and 10 had periodontitis. B. forsythus (BF), P. gingivalis (PG), P. intermedia (PI), P. nigrescens (PN), A. actinomycetemcomitans (AA), T. denticola (TD), and pathogen related oral spirochetes (PROS) were studied. Oral hygiene was similar and poor in both groups. The mean probing depth at sample sites was 6.7 mm (S.D+/-1.3) in the periodontitis group and 2.2 mm (S.D.+/-1.5) in the gingivitis group (F=17.75, p<0.001). Mean clinical attachment levels (CAL) were 4.3 mm (S.D.+/-2.0) and 1.7 mm (S.D.+/-0.9) respectively (p<0.001). Total viable counts >1.0x10(5) in supra gingival plaque samples were found in all periodontitis and in eight gingivitis subjects. 70x more black-pigmented organisms were found in supra-gingival and 185 times more in sub-gingival plaque from the periodontitis group (p<0.01). Culture data showed P. nigresecens in 10% periodontitis and 50% gingivitis subjects (p<0.03). In supra-gingival samples by the Affirm DP test, BF was present in 50% periodontitis and 60% gingivitis while culture data were negative for all subjects. PG was found in 30% periodontitis and 50% gingivitis subjects with TD in 70% periodontitis and in 30% gingivitis subjects. In the sub-gingival plaque samples 80% periodontitis and 70% gingivitis subjects had >1x10(5) anaerobes. The total count of black-pigmented organisms was significantly greater in the periodontitis elders (p<0.001). cDNA probes by the Affirm DP test identified subgingival presence of BF (80%) PG (80%), PI (80%), AA (0%), TD (50%) in periodontitis subjects with BF (70%), PG (40%), PI (30%) and TD (20%) in gingivitis subjects. PROS were found in (80%) samples from periodontitis and in (60%) of gingivitis elderly. Only the quantities of PI (r=0.48, p<0.01) and TD (r=0.37, p<0.01) were associated with the disease definition. The smoking habit in the periodontitis group was significantly higher (p<0.01). A history of smoking may contribute significantly to periodontitis in the presence of pathogens. PMID- 9846800 TI - Guided tissue regeneration in the treatment of human infrabony defects. Clinical, radiographical and microbiological results: a pilot study. AB - The aim of the study was to evaluate the clinical, radiographical and microbiological outcome after using guided tissue regeneration (GTR) with a bioabsorbable membrane, Resolut. Subjects with bilateral infrabony defects at single rooted teeth were selected. A total of 22 teeth, 2 in each 1 of 7 patients and 4 in 2 patients, with probing pocket depth > or =5 mm, 3 months after scaling, participated. At baseline, assessments of plaque and gingival indices, bleeding on probing, probing pocket depth and probing attachment level were recorded and reproducible radiographs for computer-based bone level measurements were taken. Bacterial samples were collected to investigate the presence of periodontitis-associated bacteria, e.g., Porphyromonas/Prevotella- and Fusobactrium-like micro-organisms. One tooth was randomly treated with GTR and the contralateral with an open debridement procedure as a control. Clinical, radiographical and microbiological examinations were repeated 6 and 12 months postoperatively. Both procedures demonstrated a statistically significant improvement of gingival conditions, reduction of pocket depths and gain of attachment. When evaluating the differences between test and control teeth, none of the clinical parameters yielded statistical difference. Computer-based bone level measurements showed only small differences in the majority of both test and control sites. The differences were not significant. Periodontitis-associated bacteria were present at baseline, but the appearance was not related to any specific site or patient and did not demonstrate any unwanted change in the 6- and 12-month samples. The findings suggest that the clinical, radiographical and microbiological improvements were not significantly enhanced with the GTR therapy. PMID- 9846801 TI - Associations between salivary calcium and oral health. AB - Recently, we have shown positive correlations between high salivary calcium content and periodontitis, and between high salivary calcium level and the number of intact teeth in selected groups of subjects. The aim of our present study was to determine whether these correlations could be seen in a randomized group of healthy adults. A thorough oral examination including orthopantomograms was carried out for a total of 137 healthy subjects, 63 men (35.4+/-5.6 years) and 74 women (33.2+/-4.7 years). Paraffin-stimulated saliva was collected from the subjects and salivary flow (ml/min), buffering capacity, calcium (mMol/l) and microbial variables including lactobacilli, yeasts, mutans streptococci, total streptococci, total number of aerobes, and anaerobes were determined. The calcium level of whole saliva had a median of 1.23 mMol/l. Subjects with calcium level below the median were categorized as 'low', while those with higher values formed the 'high' salivary calcium group. There were more men than women in the 'High' salivary calcium group (p=0.025). Subjects in the 'high' calcium group showed more bleeding on probing (p=0.026), had more intact teeth (p=0.045), and lower DMF-scores (p=0.025) than their counterparts. No other differences were found between the two groups. We found clear associations between the level of salivary calcium and factors reflecting gingival health on one hand, and dental health on the other in a randomly selected group of healthy subjects, and conclude that salivary calcium may be important with regard to both dental and gingival health. PMID- 9846802 TI - Expanded polytetrafluoroethylene and dental rubber dam barrier membranes in the treatment of periodontal intrabony defects. A comparative clinical trial. AB - As observed in previous case reports, dental rubber dam (DRD) can be utilized as a barrier membrane in the guided tissue regeneration (GTR) technique for the treatment of periodontal intrabony defects. The purpose of the present study was (1) to confirm the validity of DRD as a suitable material in regenerative procedures and (2) to compare, in a split-mouth clinical trial, the effectiveness of DRD-made membranes in the treatment of periodontal intrabony defects versus that of expanded polytetrafluoroethylene (e-PTFE) barriers. 22 systemically healthy non-smoker adult periodontitis patients (7 male, 15 female) aged between 35 to 58 years were selected for the study. In each patient, a couple of 2-3 wall intrabony defects, located in different quadrants, were treated by a GTR technique using DRD (test sites) and e-PTFE (control sites), respectively. Performing a strict control of the oral hygiene level and of the marginal gingival health during the whole period of study, clinical (pocket probing depth, PPD; probing attachment level, PAL; gingival recession, GR) and intrasurgical (depth of the defect's intraosseous component, IOC; level of the alveolar crest, ACL) parameters were recorded at baseline and at the 1-year re-entry procedure in each experimental site. Furthermore, the coronal level of the newly formed tissue from the base of the defect (NFTL) and the vertical bone gain (VBG) were calculated at the time of membrane removal and after the re-entry procedure respectively. Membranes were removed from both test and control sites after 5 weeks; however, exposure of the membrane always occurred in test sites whereas it was observed in only 6 out of 22 control sites, this fact leading to an incomplete coverage of the regenerated tissue by the gingival flap in 18 out of 22 test sites. In both test and control sites, a statistically significant improvement of clinical and intrasurgical parameters occurred at the end of the study period; however, a significantly greater improvement was observed in control sites for PAL (+4.0 mm versus +3.0 mm; p<0.05) and VBG (3.9 mm versus 2.9 mm; p<0.05) although at the time of membrane removal, NFTL was similar between the experimental sites (test: 5.8; control: 5.6; p>0.05). Conversely, test sites exhibited a statistically significant greater increase in gingival recession (+1.9 versus +1.2; p<0.05) and alveolar crest resorption (-1.1 versus -0.3, p<0.01) in comparison to controls. It was concluded that (1) DRD is a suitable material to be used as a barrier membrane in GTR procedures although (2) e-PTFE membranes can provide a greater improvement in PAL and VBG, probably because of the difficulty in completely covering the regenerated tissue due to the fact that the gingival tissues have undergone a consistent recession in DRD-treated sites. Further studies are needed to demonstrate if an adequate coverage of the regenerated tissue in DRD-treated sites can eliminate these differences. PMID- 9846803 TI - Presence of cortisol in gingival crevicular fluid. A pilot study. AB - Cortisol is one of the primary mediators of the stress response, in the main having immunosuppressive effects. An important component of the host response in periodontal inflammation is gingival crevicular fluid (GCF), with constituents mainly derived from serum. Cortisol, like many other steroids, is present in saliva but its occurrence in GCF does not seem to be documented. Unstimulated whole saliva was collected and GCF was sampled on filter disks. The samples were analysed by a modified RIA method for serum in such a way that small volumes and low concentrations could be measured. Our findings suggest that the total concentration of cortisol in GCF might be estimated to levels below 1/10 of that in serum. However, what appears as a distinctive feature is the considerable variation of the cortisol concentrations for individual teeth. To our knowledge, this is the first time cortisol has been measured in gingival crevicular fluid, and this opens the prospects for further in vivo research. PMID- 9846804 TI - Porphyromonas gingivalis in an edentulous proband. A case-report. AB - In a previous study, an edentulous subject showed the presence of P. gingivalis, only in saliva. The present case report is a longitudinal follow-up on this proband (female, 63 years) and her spouse with periodontitis (67 years). Samples were taken from the proband and the spouse for microbiological analysis at several occasions during a 3-year period and a 1-year period respectively. In the edentulous proband, P. gingivalis was present in saliva only, at all times. In the spouse, P. gingivalis was present in the periodontal pocket and on the mucous membranes, but the saliva was culture-negative for this species at both sampling times. The restriction enzyme analysis (REA) patterns of isolates (n=7) of P. gingivalis from the proband recovered over 3 years showed no differences. This indicated that the same clonal type of P. gingivalis persisted for 3 years in the proband. The P. gingivalis isolates (n=13) from the spouse showed no differences when REA patterns were compared. Comparison of REA patterns from the proband and the spouse also showed no differences. Thus the same clonal type of P. gingivalis can persist over several years in an edentulous subject and at least for 1 year in a subject with periodontal disease. PMID- 9846805 TI - Effect of acidosis on contraction, intracellular pH, and calcium in the newborn and adult rabbit aorta. AB - This study investigated the effect of acidosis on intracellular pH (pHi), intracellular calcium concentration ([Ca]i), and vascular contraction in the aorta of the newborn and adult rabbit. Isometric tension, pHi, and [Ca]i were measured in an isolated ring preparation. After the vascular contraction was induced with 50mM KC1, the effect of respiratory acidosis produced by elevation of PCO2 was studied. Respiratory acidosis caused a transient depression followed by a recovery of contractile tension. The decrease in developed tension was greater in the newborn than in the adult. The decrease in pHi during acidosis was similar in the two age groups. [Ca]i increased during acidosis and the increase was greater in the newborn than in the adult. These data show that the vasorelaxant effect of acidosis in the newborn aorta is greater than that in the adult aorta. The greater vasodilation in the newborn cannot be explained by the difference in pHi or [Ca]i. PMID- 9846806 TI - Change in cardiac diastolic function in neonates. AB - In this study we used Doppler echocardiography to examine changes in ventricular diastolic function in neonates. Echocardiograms were performed in 25 healthy neonates at 1-6 days (2.7+/-1.6 days) and at 25-35 days (29.4+/-3.3 days) after birth. Ventricular inflow velocity curves were recorded using the pulsed-Doppler method. Peak velocities (peak E, A). mean velocities (mean E, A), times (time E, A), and diastolic velocity-time integrals (area E, A) were measured during both early ventricular filling and atrial systole. The acceleration time of the wave during early ventricular filling was also measured. Peak E and A and mean E and A in both ventricles increased at 1 month after birth. Acceleration times in both ventricles were shortened. The ratio of area E to A in both ventricles and of peak E to A in the left ventricle did not change at 1 month after birth. Only the ratio of peak E to A in the right ventricle increased. The ratio of peak E to (area E + area A), which is thought to be a load-independent index, increased in both ventricles. The authors concluded that ventricular relaxation appears to increase in both ventricles, 1 month after birth, and that changes in diastolic function occur even in neonates. PMID- 9846808 TI - Effects of a cardioselective M2 receptor antagonist, AF-DX 116, on ventricular arrhythmias in dogs. AB - AF-DX 116 is a cardioselective M2 receptor antagonist and, thus, it should induce sinus tachycardia. Since normal ventricular automaticity is suppressed by atrial overdriving, AF-DX 116 might become an antiarrhythmic drug and act by increasing the sinus node automaticity. Ventricular arrhythmia models used in this study were induced either by two-stage coronary ligation, digitalis, or adrenaline in beagle dogs. AF-DX 116 (0.3 mg/kg i.v.) tended to increase the sinus rate of the conscious beagles compared to the pre-drug level (although the effect was not significant) but had no antiarrhythmic effect (i.e., there was no decrease in the arrhythmic ratio defined as the number of PVC divided by the total heart rate) on the ventricular tachycardia (VT) induced 24 h after aseptic ligation of the left anterior descending coronary artery. AF-DX 116 did not alter the blood pressure. The drug also did not suppress digitalis-induced VT and did not increase the atrial rate, which was already increased to about 210 beats/min by ouabain. AF-DX 116 decreased the arrhythmic ratio, 9 min after bolus injection, and increased the atrial rate in the adrenaline VT model. The maximum plasma concentration of AF-DX 116 reached nearly 1microg/ml 1 min after the bolus injections in the digitalis and adrenaline arrhythmia experiments, which is close to the maximum concentration expected to be attained in the clinical application of this drug. Although AF-DX 116 increased the heart rate, under the present experimental conditions of increased atrial rate, the extent of tachycardia was not strong enough to suppress the 24-h coronary ligation and digitalis-induced arrhythmias. The late onset of the antiarrhythmic effect of AF-DX 116 on adrenaline-induced arrhythmia cannot be explained by the overdrive suppression mechanism. PMID- 9846807 TI - Failure of treatment with interleukin-2 receptor-specific monoclonal antibody in acute coxsackievirus B3 myocarditis in mice. AB - T cell activation is assumed to play a crucial role in many viral infections. An important marker for the activation of T cells is the interleukin-2 receptor (IL 2R); resting T lymphocytes do not bear detectable amounts of IL-2R. AMT13, a rat monoclonal antibody against mouse IL-2R, inhibits interleukin-2-dependent cell growth both in vitro and in vivo. Therefore, to clarify the effects of anti-IL-2R antibody treatment upon coxsackievirus B3 (CB3)-infected C3H/He mice, AMT13, 1 microg/mouse per day, was administered, subcutaneously, starting on day 0 (group 2) in experiment I or on day 7 (group 4) in Experiment II for 7 days, respectively. Groups 1 and 3 were examined as infected controls. In both experiments, there was no significant difference in mortality or in the severity of myocarditis between the treated and the untreated groups. Also, myocardial CB3 titers on day 7 did not differ significantly between groups 1 and 2. In addition, the distribution of activated T cell subsets in the inflamed myocardium was not changed by the treatment, and the paucity of myocardial IL-2R-positive cells was confirmed in all groups. Effects of the antibody treatment were confirmed by a decrease in delayed type hypersensitivity. Although some reports have shown that anti-IL-2R antibody has been successfully applied to ameliorate acute renal graft versus-host disease, to enhance survival of skin allografts, and to suppress diabetic insulitis, it did not exert a beneficial effect on acute CB3 myocarditis in mice. PMID- 9846809 TI - Correction of erythrocyte shape abnormalities in familial hypercholesterolemia after LDL-apheresis: does it influence cerebral hemodynamics? AB - It is well known that red blood cells incubated in low-density lipoprotein (LDL) rich medium show shape abnormalities that revert to normal after reincubation in normal plasma. Patients with homozygous familial hypercholesterolemia (HFH) have an increased percentage of abnormally-shaped erythrocytes (mostly stomatocytes, knisocytes, and crenated cells) compared to normocholesterolemic controls: 7.73+/ 0.96 versus 3.52+/-0.52 (mean+/-SEM; P = 0.001). To confirm the role of high LDL concentration in inducing red cell shape abnormalities we determined the percentage of abnormally shaped erythrocytes in seven HFH patients 1 day after the procedure of LDL-apheresis with a 40% cholesterol decrease. A reduction in kniscocytes, stomatocytes, and crenated cells was observed in the patients treated by LDL-apheresis (P < 0.01). To investigate the possible benefit of a reduction in erythrocyte shape abnormality on cerebral hemodynamics, cerebral flow velocity, as evaluated by transcranial Doppler, was evaluated concomitantly and found to be remarkably increased after apheresis (P < 0.01). No significant change in hematocrit, plasma viscosity, blood viscosity, mean pressure, or cardiac output was detected, 1 day after apheresis. An inverse correlation was demonstrated (r = 0.55; P = 0.04) between changes in the percentage of knisocytes+stomatocytes +crenated cells and percent changes in middle cerebral artery peak systolic velocity. The correction of erythrocyte shape abnormalities after LDL-apheresis might be related to dramatic changes in plasma phospholipid concentration and proportion occurring after this procedure in HFH patients. The reduction of erythrocyte shape abnormalities could contribute, together with other hemorheological factors, to the improvement of cerebral hemodynamics after LDL-apheresis. PMID- 9846810 TI - Hepatocyte growth factor(HGF): a new biochemical marker for acute myocardial infarction. AB - The purpose of this study was to investigate the use of hepatocyte growth factor as a biochemical marker for acute myocardial infarction. Several biochemical markers are used for noninvasive detection of acute myocardial infarction. However, hepatocyte growth factor has not been used previously for this purpose. We measured hepatocyte growth factor, creatine phosphokinase, and MB isoenzyme (CK-MB) levels in 6 patients with stable effort angina after diagnostic catheterization (controls) and in 12 patients with acute myocardial infarction (AMI). The measurements in the AMI patients were recorded twice a day for the first 3 days after onset of chest pain and once a day for the next 4 days. Furthermore, in each patient we evaluated the time to reach the maximum level and the time for the level to decline to less than half the maximum. Hepatocyte growth factor levels (ng/ml) were 0.3+/-0.1 for angina pectoris patients, and 15.7+/-9.1 within 6h and 12.5+/-4.6 within 12h after the onset for AMI patients, respectively. The correlation coefficients between hepatocyte growth factor and creatine phosphokinase and between hepatocyte growth factor and CK-MB were 0.68 and 0.74, respectively. The time to reach the maximum (h) and the time to decline to less than half of the maximum level (days) were 6.6+/-2.6 and 1.2 +/-0.2 for hepatocyte growth factor, 19.4+/-8.7 and 2.5+/-1.4 for creatine phosphokinase, and 16.6+/-7.7 and 1.5+/-0.4 for CK-MB, respectively. Hepatocyte growth factor is useful as a prognostic indicator and reflects the clinical course in patients with acute myocardial infarction. PMID- 9846811 TI - Chylothorax in a polysplenia infant with cor triatriatum, pulmonary stenosis and sick sinus syndrome. AB - An infant with chylothorax and polysplenia is described. Her chylothorax was refractory and related to right-sided heart failure derived from pulmonary stenosis, cor triatriatum, and sick sinus syndrome. In addition to Brock operation and cardiac pacing, thoracic duct ligation was performed, but it resulted in an only transient amelioration of the chylothorax. Eventually, the chylothorax resolved when total cardiac repair was performed for cor triatriatum. We propose that, in a patient with chylothorax and right-sided heart failure caused by compound cardiac malformations, as in this patient early surgical treatment for the underlying disease of right-sided heart failure should be considered. PMID- 9846812 TI - Dicrotic pulse observed in a patient with prolapse of the aortic valve without aortic regurgitation. AB - We describe a case of dicrotic pulse in a patient who exhibited marked prolapsed aortic cusp without aortic regurgitation. Echo-Doppler in the abdominal aorta showed deep reversal flow confined to early diastole. We conclude that this prolapse was the cause of the steep dicrotic notch and the dicrotic pulse, because of the concomitance of non-leaking aortic valves. PMID- 9846813 TI - Cytokine secretion of myelin basic protein reactive T cells in patients with multiple sclerosis. AB - The objective of this study was to determine whether autoreactive T cells in patients with multiple sclerosis (MS) are polarized and committed in their differentiation to a stable cytokine phenotype or whether the cytokine secretion can be altered. We examined the cytokines secreted by myelin basic protein (MBP) as compared to tetanus toxoid-reactive (TT) T cells in 12 patients with relapsing remitting MS (RR-MS), 9 patients with chronic progressive MS (CP-MS), and 14 normal individuals. A total of 5094 short term T cell lines to MBP and TT were generated in the presence of growth conditions promoting Th1 (IL-12/alpha-IL-4 mAb) or Th2 (IL-4/alpha-IL-12 mAb) cytokine secretion. Antigen-specific cytokine secretion from normals and MS patients could be shifted to a Th1 or Th2 type phenotype depending upon culture conditions, indicating that the phenotype of MBP reactive T cells can be altered even in longstanding chronic progressive MS. There were no significant differences in the cytokine patterns secreted by MBP reactive T cells in patients with MS as compared to normal individuals. However, CP-MS patients tended to have fewer MBP reactive T cells secreting IL-4 when cultured with IL-12/anti-IL-4 mAb and more IFN-gamma secreting MBP reactive T cells when cultured with IL-4/anti-IL-12 mAb as compared to both normal controls and RR-MS, suggesting that cells from these patients might be more polarized or that fewer undifferentiated MBP-reactive cells are present in these individuals. The most striking observation was that in contrast to the RR-MS patients and normal controls, almost none of the MBP reactive T cells secreting cytokines in CP-MS incorporated 3[H]thymidine. This may be due to chronic in vivo stimulation in the presence of IL-12, or because these T cells may have entered a terminally differentiated state. Nonetheless, the ability to alter the cytokine secretion of autoreactive T cell lines even in longstanding autoimmune disease indicates that cytokine therapy might have therapeutic benefits by switching the function of myelin reactive T cells such that they are non-pathogenic. PMID- 9846814 TI - Characterization and partial purification of a novel 36 kDa peripheral myelin protein recognized by the sera of patients with neurological disorders. AB - Sera of some patients with acquired sensory neuropathy, chronic inflammatory demyelinating polyradiculoneuropathy and motor neuron disease have high titres of IgG autoantibodies to a minor human peripheral nerve glycoprotein of approximately 36 kDa. This protein cofractionated with PNS myelin and was also found in bovine and rat nerve but not in CNS myelin or other nonneural human tissues. The N-terminal sequence revealed that this protein is related to the major myelin protein P0. Monoclonal antibodies to P0 and to the carbohydrate epitope HNK-1 did not recognize the 36-kDa protein, and the human anti-36-kDa antibodies did not bind to P0. IgG binding to this protein was not abolished after periodate oxidation or deglycosylation, suggesting that the epitope recognized by the human antibodies is peptidic. Differential glycosylation did not account for the differences in the apparent molecular weight between these two proteins. Overall our results indicate that the 36-kDa protein is a variant of P0. PMID- 9846816 TI - Strain specific variation in IFN-gamma inducible lymphocyte adhesion to rat brain endothelial cells. AB - We have examined the interferon-gamma (IFN-gamma) induced increase in lymphocyte adhesion to rat brain endothelial cells (BEC) in the experimental allergic encephalomyelitis (EAE) susceptible LEW and resistant PVG strain. A significant increase in adhesion of mitogen activated lymphocytes could be demonstrated by stimulating LEW BEC with 50 U/ml IFN-gamma for 24 h. In contrast the same treatment failed to induce a significant increase in lymphocyte adhesion in the PVG strain. Flow cytometric analysis of lymphocyte integrin expression indicated a marked increase in the number of cells expressing both LFA-1 and VLA-4 following mitogen activation and was similar between the LEW and PVG strain. Depletion of LFA-1 and VLA-4 positive lymphocytes resulted in an equivalent inhibition of adhesion to BBB-EnC indicating that the adherent population was comprised predominantly of the VLA-4 + /LFA-1 + phenotype. We conclude that this strain variation in the IFN-gamma activation of BEC may be related to the disease phenotypes of these strains. PMID- 9846815 TI - Myelin basic protein reactive Th2 T cells are found in acute disseminated encephalomyelitis. AB - Acute disseminated encephalomyelitis (ADEM), a postinfectious illness of the central nervous system (CNS), is thought to be an autoimmune disease. Here, we characterized the cytokines secreted by myelin-reactive T cells generated from patients with ADEM. The frequency of MBP-reactive T cell lines was ten-fold higher in patients with ADEM compared to patients with encephalitis and normal subjects. Whereas there was no significant IFN-gamma secretion, the predominant cytokine secreted by MBP-reactive T cell lines was IL-4 in patients with ADEM. In contrast, IL-4 secretion was only rarely detected in the controls. The presence of high frequencies of MBP-reactive IL-4 secreting T cells in subjects with ADEM during their recovery phase may be similar to myelin reactive IL-4 secreting T cells observed during the spontaneous recovery of animals with EAE. PMID- 9846817 TI - CD4+ T cell repertoire on the epsilon subunit of muscle acetylcholine receptor in myasthenia gravis. AB - We have identified sequence regions of the human muscle acetylcholine receptor epsilon subunit recognized by CD4+ T cells from myasthenia gravis patients. We tested the proliferative response in vitro of blood CD4+ cells from 18 myasthenic patients and 5 controls, to individual overlapping synthetic peptides spanning the epsilon subunit sequence. All patients recognized a complex epitope repertoire. The peptides recognized by the CD4+ cells included sequence regions of the epsilon subunit that were diverged as compared to the homologous sequences of the other receptor subunits. Recognition of epitopes formed by sequence regions unique to the epsilon subunit suggests a direct role of this subunit in sensitizing the CD4+ cells. Several epsilon subunit peptides were recognized by many patients. Thus the epsilon subunit, like other acetylcholine receptor subunits, forms 'universal' CD4+ epitopes. The healthy subjects recognized some epsilon subunit peptides sporadically and at a low level. PMID- 9846818 TI - Cultured sympathetic neurons express functional interleukin-1 receptors. AB - Since recent work has identified an apoptotic pathway in sympathetic neurons that is mediated by autocrine interleukin-1 (IL-1), we investigated whether cultured sympathetic neurons possess functional IL-1 receptors. Cultured sympathetic neurons express levels of IL-1RI and IL-1RAcP mRNAs consistent with signal transduction. Neurons stimulated with IL-1 demonstrate enhanced p65 NF-kappaB nuclear translocation and enhanced NF-kappaB DNA binding activity, with at least p65 and p50 subunits participating in the DNA binding activity. RNA differential display identified several neuronal mRNAs regulated by IL-1, including a member of the reticulon family. We conclude that IL-1 stimulates a potential component of a neuronal secretory pathway. PMID- 9846819 TI - Characterization of kappa-opioid receptor transcripts expressed by T cells and macrophages. AB - We have found that the immature T cell lines R1.1 and DPK and the macrophage lines P388D1 and WEHI-3 also express kappa-opioid receptor (KOR) mRNA. Characterization of the KOR transcripts in both brain tissue and these T cells has revealed both the normal full-length as well as a truncated form of the mRNA. Our results show that the truncated transcript lacks the second exon. Primary macrophages express this truncated form of the transcript in the absence of detectable levels of the full-length form. These results suggest a degree of heterogeneity in the expression of the opioid receptors which has not previously been reported. PMID- 9846820 TI - Antibodies to human myelin proteins and gangliosides in patients with acute neuroparalytic accidents induced by brain-derived rabies vaccine. AB - Antibody responses to myelin antigens were analysed in 15 patients who developed acute neuroparalytic accidents (ANPA) during post-exposure rabies vaccination using a rabies vaccine prepared on brain tissues and in 30 individuals who were uneventfully vaccinated. High titers (> or = 100) of IgG and IgM antibodies to GM1 or GD1a gangliosides were detected by enzyme linked immunosorbent-assay (ELISA) in plasmas from ANPA patients but not in controls. These data suggest that antibodies to GM1 and GD1a gangliosides may play a pathogenic role in the demyelinating and/or inflammatory processes characteristic of rabies vaccine induced acute neurologic complications. PMID- 9846821 TI - Immunomodulatory effects of interferon-beta-1b in vivo: induction of the expression of transforming growth factor-beta1 and its receptor type II. AB - The mechanisms by which interferon-beta-1b (IFNbeta-1b) acts in the treatment of patients with multiple sclerosis (MS) are not completely known. A total of 10 MS patients were treated with 8 million units of IFNbeta-1b every other day. Compared to baseline and control group the expression of TGFbeta-1-mRNA by PBMC was persistently increased at week 6, month 3 and month 6 (p < or = 0.04), that of the TGFbeta-1 receptor type II from day 5 up to month 6 (p < 0.01). The mRNA and protein expression of tumor necrosis factor-alpha (TNFalpha)-receptor (55 kDa) was only temporarily elevated at the beginning of the therapy. Serum levels of sVCAM were increased during the whole time of treatment (p < 0.01). The CD8CD38 lymphocyte subpopulation was continuously elevated from day 5 up to month 6 (p < 0.01). No persistently significant changes were demonstrable concerning the percentage of total CD4, CD8, CD19 or in CD4 subpopulations (CD4CD29, CD4CD45RA). The present data suggest that IFNbeta-1b induces the expression of TGFbeta-1- and TGFbeta-R-II-mRNA by PBMC and increases levels of sVCAM-1 and of circulating activated CD8 cells (CD8CD38) in serum. These might be other mechanisms by which IFNbeta-1b mediates its positive effects in the treatment of MS patients. PMID- 9846822 TI - Antibodies against beta1 and beta2 adrenergic receptors in myasthenia gravis. AB - Patients with myasthenia gravis have antibodies and T cells that react with the beta1- and beta2-adrenergic receptors. These receptors, as well as other auto antigens, are present on cardiomyocytes, skeletal muscle cells and lymphocytes and are of importance for the regulation of the functions of these organs. Antibodies against the beta1-adrenergic receptor have been implicated in dilated cardiomyopathies. Myasthenia gravis (MG) patients have been suggested to have a higher than normal prevalence of heart disease. We have analysed the isotypes, subclasses, and binding sites of the beta-adrenergic receptors antibodies in both MG patients and healthy individuals and the correlation between beta-adrenergic receptors antibodies and heart disease in MG patients. The patients have IgG antibodies that react with both beta1- and beta2-adrenergic receptors. The subclasses were predominantly IgG2 and IgG4. By using synthesised overlapping peptides representing the immunodominant regions on the receptors, it was shown that the antibodies bound to partially overlapping sites on both beta1- and beta2 adrenergic receptors, but not to peptides from the acetylcholine receptor. beta adrenergic receptor antibodies were found in 34/125 MG patients. Seven out of these 34 patients had symptomatic heart disease, all seven were over 70 years of age and had arteriosclerotic heart disease. There was no difference in the prevalence of clinical heart disease in patients with and without beta-adrenergic receptor antibodies. However, patients with heart disease had significantly higher levels of antibodies than healthy individuals and other patients. Antibodies against beta-adrenergic receptors in patients with myasthenia gravis binds to both beta1- and beta2-adrenergic receptors and might be implicated in the few patients with myasthenia gravis who have heart disease. PMID- 9846823 TI - Development of HIV encephalitis in AIDS and TNF-alpha regulatory elements. AB - Tumor necrosis factor-alpha (TNF-alpha) appears to play an important role in HIV encephalitis (HIVE). TNF2, a polymorphism of TNF-alpha, associates with higher levels of TNF-alpha and severe manifestations of some infections. We studied 44 acquired immunodeficiency syndrome (AIDS) patients with autopsy-proven HIVE and/or HIV leukoencephalopathy (HIVLE) (HIVE/LE) and 30 AIDS patients without HIVE/LE. TNF2 did not associate with presence of HIVE/LE (p > 0.5). Moreover, the TNF-alpha regulatory element TTATTTAT within the 3'-untranslated region was intact in HIVE/LE brains, and HLA-DR3 did not associate with HIVE/LE. Other host factors or, more likely, viral factors may be responsible for the development of HIVE/LE. PMID- 9846824 TI - Differential induction of interleukin-12, interleukin-18, and interleukin-1beta converting enzyme mRNA in experimental autoimmune encephalomyelitis of the Lewis rat. AB - Experimental autoimmune encephalomyelitis (EAE) is a model of autoimmune central nervous system (CNS) disease that is mediated by autoreactive Th1 cells secreting the proinflammatory cytokine interferon (IFN)-gamma. Interleukin (IL)-12 in its heterodimeric p35/p40 isoform and the recently described cytokine IL-18 potently induce T cell production of IFN-gamma. Interleukin-1beta converting enzyme (ICE) is required to convert IL-18 precursor protein into its biologically active mature form. In this study, we used semiquantitative reverse transciptase polymerase chain reaction to determine steady state levels of IL-12, IL-18, and ICE mRNA in the spinal cord of Lewis rats at different stages of EAE. In control rats, we found significant IL-18, ICE, and IL-12p35, but not IL-12p40 mRNA expression. IL-18 mRNA increased during the acute stage of EAE together with a marked induction of ICE mRNA. IL-12p35 mRNA levels did not change significantly throughout the course of EAE. Surprisingly, the peak expression of IL-12p40 mRNA was delayed by several days relative to the peak of T cell infiltration and IFN gamma mRNA synthesis. Our data implicate the IL-18/ICE pathway in the amplification of Th1-mediated immune responses in the CNS but suggest a different, so far undefined role of endogenous IL-12 in the late effector phase of EAE. PMID- 9846825 TI - Cerebral spinal fluid lymphocytes are part of the normal recirculating lymphocyte pool. AB - We have investigated the migration of lymphocytes from blood into the central nervous system (CNS) under normal physiological conditions. Using sheep as our model, we simultaneously sampled blood, lymph and cerebral spinal fluid (CSF). Normal, nonactivated, recirculating lymphocytes can migrate into the CSF in similar concentrations as found in subcutaneous lymph and there is no difference in the temporal appearance between them. Lymphocytes infused into the CNS could be found in cervical lymph nodes. These data suggest that lymphocytes found in the CNS are part of the recirculating lymphocyte pool and do not require activation to enter the CSF. PMID- 9846826 TI - No evidence for elevated numbers of mononuclear cells expressing MCP-1 and RANTES mRNA in blood and CSF in multiple sclerosis. AB - The perivascular accumulation of mononuclear cells (MNC) in brain white matter is critical in the development of active lesions in multiple sclerosis (MS). Chemokines contribute to leukocyte recruitment by increasing the adhesiveness of integrins expressed on leukocytes and by promoting migration through endothelium and extracellular matrix. By using an in situ hybridization technique, it was possible to enumerate blood and CSF MNC expressing mRNA for the two CC chemokines monocyte chemoattractant protein-1 (MCP-1) and RANTES (regulated upon activation, normal T cells, expressed and secreted) in MS patients and controls. No differences in numbers of blood MNC expressing MCP-1 or RANTES could be found in MS patients compared to healthy individuals or patients with acute aseptic meningoencephalitis (AM). High numbers of CSF MNC expressing MCP-1 and RANTES were found in some MS patients, but also in patients with AM. This shows that elevated numbers of MCP-1 and RANTES mRNA expressing CSF MNC are not specific for the inflammatory process in MS. We conclude that there is no evidence for a systemic dysregulation of the CC chemokines MCP-1 and RANTES in MS. PMID- 9846827 TI - Immune consequences of stroke and cerebral palsy in adults. AB - The possibility that brain damage results in a sustained dysregulation of lymphocyte responsiveness to the lymphokine, interleukin-2 (IL-2), was investigated in individuals who had experienced a unilateral stroke in adulthood or who presented with spastic hemiparesis since childhood. Following verification of unilateral brain damage via neuromotor assessment, and determination of their health status, blood samples were obtained to evaluate a panel of immune measures. Soluble interleukin-2 receptor (sIL-2R) and lymphocyte proliferative and cytolytic responses in the subjects with stroke or cerebral palsy were compared to age- and gender-matched controls. In addition, lymphocyte populations were enumerated via flow cytometry, and lymphocyte cyclic AMP (cAMP) levels were determined. Circulating blood levels of sIL-2R were significantly elevated in all individuals that had experienced unilateral brain damage. Cytolytic activity also failed to be stimulated to the normal level by in vitro treatment of lymphocytes with IL-2. Further, lymphocytes from the stroke subjects proliferated significantly less after mitogen and IL-2 stimulation. These functional differences were not accounted for by an abnormal leukocyte profile, although phenotypic analyses revealed subtle differences in the natural killer cell subsets. Overall, the findings indicate that individuals with brain damage may not respond appropriately when immune activation is required. These immune differences appear to be a stable trait given that they were manifested after both perinatal and adult brain insult in otherwise healthy, independently living individuals. PMID- 9846828 TI - Identification of a delta isoform of preprotachykinin mRNA in human mononuclear phagocytes and lymphocytes. AB - We have characterized preprotachykinin (PPT-A) gene transcript splicing products and identified a fourth isoform of PPT-A mRNA transcript in human peripheral blood-isolated monocytes and PBL. Using RT-PCR, Southern blot analysis and nucleotide sequencing analysis, we have identified the four isoforms of PPT-A transcripts (alpha, beta, gamma and delta) in human peripheral blood-isolated monocytes and PBL. The delta-PPT transcript present in the immune cells lacks exons 4 and 6. The sequences of exons 3, 5 and 7 of delta-PPT transcript completely match those of beta-PPT transcript. The alpha-PPT and beta-PPT sequences in these cells are identical to those obtained by Tan and Too (GenBank accession number U37539) and Harmar et al. (Genbank accession number X54469), but differ by a single nucleotide from another entry by Chiwakata et al. (Genbank accession number M68906). In comparison to this latter sequence, there was a C- >T change at amino acid position 87 (CCT-->CTT) which may result in a Pro to Leu change. Identification of the new SP mRNA transcript in both human CNS and immune cells supports the concept of an important biological link between CNS and immune system. PMID- 9846829 TI - T cell receptor beta-chain repertoire in inclusion body myositis. AB - Inclusion body myositis (IBM) is the most common muscle disease affecting individuals over 50 years of age. The inflammatory reaction is characterized by cell infiltrates predominated by CD8+ cytotoxic T cells. To analyze clonality of muscle infiltrating lymphocytes, we studied the complementarity determining region 3 (CDR3) length distribution of the T cell receptor (TCR). Muscle infiltrating lymphocytes were studied in three IBM patients and compared with peripheral blood lymphocytes (PBL) in two of these patients. The study was performed by reverse transcription polymerase chain reaction (RT-PCR) of RNA extracted from muscle tissue and PBL followed by analysis of fragment length distribution of the CDR3 region in each of 24 different Vbeta families. There was a restricted usage of TCR Vbeta gene families in muscle infiltrating T cells in all three patients. Some of the TCR Vbeta gene families showed oligoclonal expansions but polyclonal patterns were dominating. The CDR3 distribution of most Vbeta families differed between muscle infiltrating lymphocytes and PBL indicating that T cells have expanded locally or selectively accumulated in muscle. PMID- 9846830 TI - Bystander suppression of experimental autoimmune encephalomyelitis by T cell lines and clones of the Th2 type induced by copolymer 1. AB - The synthetic amino acid copolymer, copolymer 1 (Cop 1) induces T suppressor (Ts) lines/clones, which are confined to the Th2 pathway, cross react with myelin basic protein (MBP), but not with other myelin antigens on the level of Th2 cytokine secretion. Nevertheless, Cop 1 Ts cells inhibited the IL-2 response of a proteolipid protein (PLP) specific line. Furthermore, Cop 1 Ts cells ameliorated EAE induced by two unrelated encephalitogenic epitopes of PLP: p139-151 and p178 191, that produced different forms of disease. This bystander suppression demonstrated by the Cop 1 Ts cells may explain the therapeutic effect of Cop 1 in EAE and MS. PMID- 9846831 TI - Kinetics of anaphylatoxin C5a receptor expression during experimental allergic encephalomyelitis. AB - In this study, we investigated the expression of the C5aR in spinal cords of Lewis rats with experimental allergic encephalomyelitis (EAE). Using in situ hybridization (ISH) we analyzed the kinetics of C5aR at different time points of EAE (preclinical stage, clinical peak, remission phase). We observed that C5aR mRNA was readily detected in the CNS of EAE rats at all the stages of the disease. Using a combination of ISH and immunohistochemistry, we formally demonstrated that C5aR is strongly expressed on microglial cells and hypertrophic astrocytes during EAE. The potential involvement of C5a receptor in EAE physiopathology is discussed. PMID- 9846832 TI - Treatment of murine experimental autoimmune encephalomyelitis with a myelin basic protein peptide analog alters the cellular composition of leukocytes infiltrating the cerebrospinal fluid. AB - Experimental autoimmune encephalomyelitis (EAE) can be effectively treated during disease exacerbation by administration of a peptide corresponding to the major T cell epitope of myelin basic protein (MBP), but the mechanism by which T cell tolerance leads to clinical improvement is not well-defined. Acute exacerbations of EAE are accompanied by an infiltration of blood-borne leukocytes into the brain and spinal cord, where they mediate inflammation and demyelination. To investigate peptide effects on infiltrating cells, we collected cerebrospinal fluid (CSF) from (PL/JxSJL)F1 mice with MBP-induced EAE. Pleiocytosis by lymphocytes, neutrophils, and macrophages was seen throughout the course of relapsing-remitting disease. A single administration of the MBP peptide analog, Ac1-11[4Y], reduced disease severity, accompanied by a dramatic and selective loss of neutrophil pleiocytosis. A longer course of peptide therapy resulted in complete recovery from clinical signs of disease, and decreased pleiocytosis by all cell types. Clinical severity throughout the course of disease and therapy was directly related to the degree of infiltration by neutrophils and macrophages, and the clinical improvement following peptide therapy was accompanied by decreased central nervous system (CNS) expression of chemoattractants for these cell types. These observations support a model of disease exacerbation mediated by phagocytic cellular infiltration under the ultimate control of T cell-derived factors, amenable to treatment by down regulation of the T cell activation state. PMID- 9846833 TI - Involvement of capsaicin sensitive primary afferents in thymulin-induced hyperalgesia. AB - Intraplantar (5 ng) or intraperitoneal (50 ng) injections of thymulin, produced both thermal and mechanical hyperalgesia in rats. In this report, we show that ablation of capsaicin sensitive primary afferents (CSPA) can alter or abolish thymulin-induced hyperalgesia. Different groups of rats were subjected to either treatment with capsaicin or to surgical subdiaphragmatic vagotomy (SDV). Both capsaicin and SDV reduced significantly thymulin-induced hyperalgesia. On the other hand, these treatments elicited differential effects on the modulation by thymulin of the levels of nerve growth factor and interleukin 1beta. We conclude that the hyperalgesic effects of i.p. thymulin are mainly mediated through the CSPA fibers. PMID- 9846834 TI - Complement activation and CD59 expression in the motor facial nucleus following intracranial transection of the facial nerve in the adult rat. AB - Intracranial transection of the facial nerve has been shown to cause a massive neuronal cell death in the motor facial nucleus. Complement activation has been proposed to contribute to neuronal degeneration following axotomy. Using immunocytochemistry and in situ hybridization we show in the present study that there is complement activation in the facial nucleus after intracranial facial nerve transection as well as increase of the complement regulators CD59 and clusterin. We propose a neuroprotective role for the complement regulators CD59 and clusterin against homologous attack of complement to facial motor neurons. PMID- 9846835 TI - Astrocyte expression of monocyte chemoattractant protein-1 is differentially regulated by transforming growth factor beta. AB - The pathophysiology of central nervous system (CNS) inflammatory disease is dependent, in part, on leukocyte recruitment across the blood-brain barrier. The expression of cytokines and chemokines by astrocytes may contribute to this process. Astrocytes express monocyte chemoattractant protein-1 (MCP-1), an activator of monocytes and a chemoattractant for monocytes and activated T cells. We examined the regulation of MCP-1 expression in human fetal astrocytes following cytokine treatment in the presence and absence of transforming growth factor beta (TGF-beta). TGF-beta, TNFalpha and IL-1beta, but not IFNgamma, induced MCP-1 mRNA and protein. TGF-beta, in cotreatment with TNFalpha caused an additive increase in MCP-1 mRNA, but not protein. In combination with IFNgamma, TGF-beta significantly increased MCP-1 mRNA and protein, as compared to either untreated, TGF-beta- or IFNgamma-treated astrocytes. However, TGF-gamma in cotreatment with IL-1beta decreased MCP-1 mRNA and protein, as compared to IL 1beta alone. Treatment of astrocytes with TGF-beta prior to TNFalpha, IFNgamma or IL-1beta treatment significantly increased MCP-1 expression. The kinetics of cytokine expression in the CNS may differentially regulate astrocyte-derived MCP 1 expression and subsequent recruitment and activation of leukocytes. PMID- 9846836 TI - B7-1 (CD80), B7-2 (CD86), interleukin-12 and transforming growth factor-beta mRNA expression in CSF and peripheral blood mononuclear cells from multiple sclerosis patients. AB - Costimulatory molecules B7-1 (CD80) and B7-2 (CD86) are differently involved in T cell stimulation. In chronic experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis (MS), B7-1 was preferentially involved in pathophysiology of relapses. We used reverse transcription polymerase chain reaction (RT-PCR) to amplify the mRNA coding for these molecules in cerebrospinal fluid (CSF) cells and peripheral blood mononuclear cells (PBMC) from 18 MS patients and 21 other neurological patients. In CSF cells of MS cases, B7-1 mRNA was only detected in some patients who showed clinical signs of acute relapse at the time of the spinal tap, while B7-2 mRNA was widely detectable without difference between active or stable MS and controls. mRNA coding for transforming growth factor-beta (TGF-beta) was detectable in the majority of cases, with higher expression in CSF cells of MS and other inflammatory neurological diseases (OIND) than in noninflammatory controls, and higher expression in PBMC of MS patients than in all other cases. Finally, mRNA coding for interleukin (IL)-12p40 was only detected in a very few number of MS and inflammatory cases. These findings were related to previous detection of other cytokines in the same cases, showing relationships in CSF cells between high expression of B7-1, IL-12p40 and TNF-alpha. PMID- 9846837 TI - Serum antibodies to heparan sulfate glycosaminoglycans in Guillain-Barre syndrome and other demyelinating polyneuropathies. AB - We tested for serum antibodies to glycosaminoglycans (GAGs), including heparan sulfate, in patients with Guillain-Barre syndrome (GBS) and other disorders. We used ELISA methods that optimize immunoglobulin binding to carbohydrate antigens to measure serum antibodies to heparan sulfate GAGs in GBS, and control neuromuscular and immune disorders. We found serum IgM or IgG antibodies to heparan sulfate GAGs in 34% of patients with GBS. Serum IgM binding to heparan sulfate GAGs was also found in some chronic demyelinating polyneuropathies, with the highest frequency (33%) in patients with IgM anti-MAG M-proteins. Antibodies to heparan sulfate GAGs were rare (1%) in control serums from patients with other disorders. This result is the first demonstration of high titer serum antibodies to a specific antigen in a substantial group of, and with some specificity for, patients with the classically described GBS syndrome of acute-onset, motor sensory polyneuropathy with demyelinating features. PMID- 9846838 TI - Dietary fat and obesity: an unconvincing relation. PMID- 9846839 TI - Does apolipoprotein E genotype influence dietary modification of circulating cholesterol concentrations? PMID- 9846840 TI - Calcium requirements: the need to understand racial differences. PMID- 9846841 TI - Is dual-energy X-ray absorptiometry ready for prime time in the clinical evaluation of body composition? PMID- 9846842 TI - Dietary fat intake does affect obesity! AB - There is a difference of opinion about whether the percentage of dietary fat plays an important role in the rising prevalence of overweight and in its treatment once it has developed. We believe that ample research from animal and clinical studies, from controlled trials, and from epidemiologic and ecologic analyses provides strong evidence that dietary fat plays a role in the development and treatment of obesity. A reduction in fat intake reduces the gap between total energy intake and total energy expenditure and thus is an effective strategy for reducing the present epidemic of obesity worldwide. A review of the results from 28 clinical trials that studied the effects of a reduction in the amount of energy from fat in the diet showed that a reduction of 10% in the proportion of energy from fat was associated with a reduction in weight of 16 g/d. We thus conclude that dietary fat plays a role in the development of obesity. To reduce the prevalence of obesity, there must be an increase in energy expenditure, a reduction in total energy intake, or both. This goal can be facilitated by reducing the amount of fat in the diet. PMID- 9846843 TI - Survey of clinical nutrition training programs for physicians. AB - Clinical nutrition training programs for physicians were surveyed to determine their number, demographic characteristics, primary teaching focuses, number of available trainee positions, funding bases, trainee numbers, backgrounds, and career positions taken. Twenty-two active programs were identified, compared with 38 programs in 1993. Thirteen of the programs were primarily focused on adult nutrition and 7 were focused on pediatric nutrition. Twelve programs appeared to have nutrition as their sole subspecialty focus, 8 were housed within gastroenterology fellowships, and 2 were within endocrinology fellowships. Most programs included training in research, which is conducted during a second or third year, or both. The decrease in numbers of programs appears to have resulted not only from relocation, retirement of key faculty members, and loss of training grants, but also because of the clearer definition of clinical nutrition training programs in this survey. The changes also reflect a national trend toward decreasing subspecialization. Within this climate, it is apparent that a new model for the training and career activities of physician nutrition specialists is needed that will attract more physicians into the discipline of nutrition. Intersociety efforts are underway to address this need and to develop a unified voice that can guide clinical nutrition training for physicians into the 21st century. PMID- 9846845 TI - Chronic ingestion of lycopene-rich tomato juice or lycopene supplements significantly increases plasma concentrations of lycopene and related tomato carotenoids in humans. AB - The bioavailability of lycopene from tomato juice and 2 dietary supplements, each containing 70-75 mg lycopene, was studied in 15 healthy volunteers in a randomized, crossover design. Subjects ingested lycopene-rich tomato juice, tomato oleoresin, lycopene beadlets, and a placebo for 4 wk each while consuming self-selected diets. Treatment periods were separated by 6-wk washout periods. Plasma lycopene concentrations, assessed at baseline and weekly throughout the treatment periods, were significantly higher during tomato juice, oleoresin, and lycopene beadlet ingestion than during placebo ingestion. Mean (+/-SEM) increases in plasma lycopene at week 4 of tomato juice, oleoresin, and lycopene beadlet ingestion were not significantly different: 0.24 +/- 0.07, 0.23 +/- 0.05, and 0.24 +/- 0.06 micromol/L, respectively. Plasma concentrations of phytofluene and phytoene, which were present in small amounts in tomato juice, oleoresin, and lycopene beadlets, increased significantly with ingestion of these 3 products. Beta-carotene, zeta-carotene, and 2,6-cyclolycopene-1,5-diol (a metabolite of lycopene)--also present in tomato juice and supplements--were significantly increased with consumption of the tomato juice and lycopene beadlets, but not with oleoresin consumption. A marked increase in plasma concentrations of an unknown compound was observed; it was detected in trace amounts in tomato juice, oleoresin, and lycopene beadlets, and had a maximum absorbance at 448 nm and a molecular weight of 556. Concentrations of plasma lycopene and other carotenoids with potential for enhancing human health can be increased by ingestion of realistic amounts of tomato juice. Lycopene appears to be equally bioavailable from tomato juice and the supplements used in this study. PMID- 9846844 TI - Thermogenic effects of sibutramine in humans. AB - BACKGROUND: Sibutramine is an effective compound for the treatment of obesity, acting both on serotonergic and noradrenergic pathways. Animal studies have shown that sibutramine exerts its effect by enhancing satiety as well as by increasing thermogenesis. OBJECTIVE: We tried to compare the acute thermogenic effect of a single 30-mg dose of sibutramine with placebo on basal energy expenditure (EE) and diet-induced thermogenesis. DESIGN: The study was randomized, double-blind, and placebo controlled. Eleven healthy, normal-weight men underwent 4 distinct treatment regimens separated by washout periods of 6-10 d. EE was measured by indirect calorimetry before and for 5.5 h after sibutramine or placebo administration with or without a 2.1-MJ breakfast. Visual analogue scales for assessment of appetite were completed hourly. RESULTS: Sibutramine caused a significant increase in EE above that for placebo (over 5.5 h) during both the fed (34%, 0.15 kJ/min) and fasted (183%, 0.20 kJ/min) states (P < 0.02) as well as during the last 3.5 h of this 5.5-h period and in the fed (87%, 0.26 kJ/min) and fasted (152%, 0.22 kJ/min) states, respectively (P < 0.01). The sibutramine induced increase in EE was accompanied by an increase in plasma epinephrine (P < 0.01), heart rate (P < 0.001), blood pressure (P < 0.05), and plasma glucose (P < 0.02). About 25% of the increased heart rate with sibutramine could be explained by increased thermogenesis. Sibutramine increased satiety more than did placebo (5-h area under the curve, P < 0.05). CONCLUSIONS: Sibutramine caused a significant increase in both EE and satiety, which may both contribute to its weight-reducing properties. PMID- 9846846 TI - Effect of a stearic acid-rich, structured triacylglycerol on plasma lipid concentrations. AB - BACKGROUND: Structured lipids are being incorporated into foods to reduce their energy value. One such lipid is rich in stearic acid. OBJECTIVE: The objective of this study was to compare the effects on plasma lipids of a stearic acid-rich triacylglycerol and a fat rich in palmitic acid in hypercholesterolemic subjects. DESIGN: Fifteen subjects with an average plasma cholesterol concentration of 6.13 +/- 0.80 mmol/L initially ate a low-fat diet for 2 wk (run-in period), followed in random order and blinded fashion by 2 high-fat diets (for 5 wk each) containing foods derived from margarines rich either in palmitic acid or in the structured, stearic acid-rich triacylglycerol. RESULTS: Plasma cholesterol concentrations with the low-fat, the stearic acid-rich, and the palmitic acid rich diets were not significantly different (5.35 +/- 0.83, 5.41 +/- 0.78, and 5.52 +/- 0.68 mmol/L, respectively) but were significantly lower (P < 0.001) than those measured during the habitual diet period (ie, 2 wk before the study began). Neither HDL cholesterol nor plasma triacylglycerol differed significantly among the 3 study diets. CONCLUSION: A similar increase in the intake of stearic and palmitic acids (differing by approximately 5% of total energy) to ensure a high fat intake resulted in plasma total and LDL-cholesterol concentrations that did not differ significantly from concentrations measured during a period of low-fat intake. PMID- 9846847 TI - Postprandial effects of an oleic acid-rich oil compared with butter on clotting factor VII and fibrinolysis in healthy men. AB - BACKGROUND: Factor VII coagulant activity (FVII:c) is associated with an increased risk of fatal ischemic heart disease, is correlated with plasma triacylglycerol concentration, and increases after a meal rich in long-chain fatty acids. OBJECTIVE: We planned to compare effects of meals rich in oleate and butter fat with those of a low-fat meal on FVII:c and fibrinolytic activity. DESIGN: A crossover design was used to compare the postprandial effects on coagulant and fibrinolytic activities in 12 men of 3 high-fat (95 g) meals--high oleate, butter, and oleate + medium-chain triacylglycerols (oleate+MCT)--with an isoenergetic low-fat meal (18 g MCT). The oleate+MCT blend was used to mimic the ratio of long-chain to shorter-chain fatty acids in butter. RESULTS: Neither the amount nor type of fat consumed influenced plasminogen activator inhibitor 1 or t plasminogen activator activities or D-dimer concentration. FVII:c increased by 12.5% (95% CI: 4.6%, 20.5%) after the high-fat meals at 3 h and by 6.7% (95% CI: 1.6%, 11.7%) at 7 h and changed 7 h after the low-fat meal by -14.3% (95% CI: 3.3%, -25.4%). The responses to the high-fat meals did not differ. Measurements of activated FVII (FVIIa), FVII zymogen, and activated FXII (FXIIa) concentrations made after the low-fat and high-oleate meals showed a significant increase in FVIIa only after the high-oleate meal. CONCLUSIONS: The results of this study confirm that FVII:c falls after a low-fat meal and suggests that postprandial activation of FVII occurs rapidly after a fat-rich meal without involving an increase in FXIIa. PMID- 9846848 TI - Activity in vitro of resveratrol on granulocyte and monocyte adhesion to endothelium. AB - BACKGROUND: Resveratrol is a phytoalexin present in red wine. It has been shown to protect LDL from peroxidative degradation. OBJECTIVE: In consideration of the low plasma concentration of orally adsorbed resveratrol (which is insufficient for antioxidant protection of LDL), we studied another effect of the compound. DESIGN: Because resveratrol is a tyrosine kinase inhibitor like other members of the tyrphostin family, we hypothesized that it has the ability to modify intracellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) expression by stimulated endothelial cells. We studied the ability of resveratrol to inhibit such adhesion molecule expression and to block the adhesion of monocytes and granulocytes to endothelial cells. RESULTS: We showed that resveratrol, at concentrations as low as 1 micromol/L and 100 nmol/L, significantly inhibited ICAM-1 and VCAM-1 expression by tumor necrosis factor alpha (TNF-alpha)-stimulated human umbilical vein endothelial cells and lipopolysaccharide-stimulated human saphenous vein endothelial cells (HSVEC), respectively. In addition, we showed that resveratrol induced a significant inhibition in the adhesion of U937 monocytoid cells to lipopolysaccharide stimulated HSVEC. Such inhibition was comparable with that obtained when anti VCAM-1 monoclonal antibody was used instead of resveratrol. Resveratrol also significantly inhibited the adhesion of neutrophils to TNF-alpha-stimulated NIH/3T3 ICAM-1-transfected cells, whereas neutrophils activated by formyl methionyl-leucyl-phenylalanine did not significantly modify adhesion to NIH/3T3 ICAM-1-transfected cells. CONCLUSIONS: Our results indicate activity of resveratrol on endothelial cells and a new interpretation of an effect independent of its antioxidant function. PMID- 9846850 TI - Lower daily energy expenditure as measured by a respiratory chamber in subjects with spinal cord injury compared with control subjects. AB - BACKGROUND: This study was designed to determine the effect of chronic spinal cord injury on daily energy expenditure. OBJECTIVE: We hypothesized that both resting and total energy expenditure would be lower in spinal cord-injured (SCI) subjects than in control subjects because of lower sympathetic nervous system activity and reduced levels of physical activity in SCI subjects. DESIGN: Twenty four-hour energy expenditure (24-h EE), resting metabolic rate (RMR), sleeping metabolic rate, spontaneous physical activity, the thermic effect of food (TEF), and 24-h respiratory quotient were measured by using a respiratory chamber in 10 male SCI subjects (injury ranged from level C6 to L3) and 59 age-matched, noninjured, male control subjects. RESULTS: The 24-h EE was lower in SCI than in control subjects (7824 +/- 305 compared with 9941 +/- 188 kJ, P < 0.01). After adjustment for fat-free mass, fat mass, and age, 24-h EE was still lower (-753 kJ/d, P < 0.01) in SCI than in control subjects. Spontaneous physical activity measured by a radar system was also significantly lower (4.6 +/- 0.6% compared with 6.5 +/- 0.3% of time, P < 0.01) in SCI than in control subjects. In absolute value (7347 +/- 268 compared with 9251 +/- 1326 kJ/d, P < 0.01) or after adjustment for fat-free mass, fat mass, and age (-678 kJ/d, P < 0.01), RMR was also lower in SCI than in control subjects. TEF was significantly lower in SCI than in control subjects (987 +/- 142 compared with 1544 +/- 213 kJ/d, representing 12.9% and 15.9% of total energy intake, respectively, P < 0.05). The sleeping metabolic rate and 24-h respiratory quotient did not differ significantly between groups. CONCLUSIONS: The 24-h EE was significantly lower in SCI than in control subjects. This difference can be explained by the lower levels of physical activity, and lower RMR and TEF values, in SCI subjects. PMID- 9846849 TI - Effect of apolipoprotein E polymorphism on serum lipid response to the separate modification of dietary fat and dietary cholesterol. AB - BACKGROUND: The magnitude of the influence of the apolipoprotein (apo) E genotype on the lipid response to different cholesterol-lowering diet modifications has been controversial. OBJECTIVE: The aim of the study was to investigate the effect of apo E genotype on serum lipid response to the separate modification of dietary fat and cholesterol. DESIGN: A prospective study design with the 3 main apo E genotype groups (3/3, 3/4, and 4/4; n = 15 in each group) was used. Groups were matched for sex, age, body mass index, menopausal status, and baseline serum cholesterol concentration. Subjects followed 3 different diets in fixed order: 1) a standardized baseline diet (38% fat, 300 mg cholesterol/d), 2) a modified National Cholesterol Education Program (NCEP) diet (34% fat, 265 mg cholesterol/d), and 3) the modified NCEP diet + cholesterol (566 mg cholesterol/d). Subjects were middle-aged (50.9 +/- 8.0 y) and mildly hypercholesterolemic (6.55 +/- 1.05 mmol/L). RESULTS: The genotype groups differed in their total cholesterol response to the NCEP diet; the mean (95% CI) decrease being greatest, -14.1% (-19.8%, -8.6% ), in subjects with apo E genotype 4/4 (P = 0.03, analysis of variance). The increase in total cholesterol after addition of 300 mg cholesterol was also greatest in subjects with apo E 4/4 [10.4% (5.8%, 15.1%)] (P = 0.03, analysis of variance). CONCLUSIONS: Apo E genotype modified the lipid response to changes in both dietary fat and cholesterol in mildly hypercholesterolemic subjects; the response was greatest in subjects with apo E genotype 4/4 and even a moderate increase in dietary cholesterol resulted in a 10% elevation in serum total cholesterol in them. PMID- 9846851 TI - Effect of protein source and quantity on protein metabolism in elderly women. AB - To study sequentially the effect of meal feeding and the effect of protein source and quantity on whole-body protein metabolism, 12 elderly women consumed 3 diets differing in both the quantity and source of protein (diet A: 5.3% of energy intake provided by animal protein and 5.0% by vegetable protein; diet B: 14.5% of energy provided by animal protein and 5.1% by vegetable protein; diet C: 5.0% of energy provided by animal protein and 15.1% by vegetable protein). The diets were consumed for 2 wk with a 2-wk interval between diets. At the end of each dietary period, nitrogen balance and protein turnover were measured. Protein turnover was measured during 4 h of fasting followed by 4 h of feeding. Comparisons were made between fasted and fed periods (within one diet) and between the diets to study the effect of the protein source and quantity. Mean nitrogen balance did not differ significantly from zero during diets B and C and was not affected by the protein source. Meal feeding resulted in increased protein flux and protein oxidation and decreased protein breakdown compared with the postabsorptive values; there was no effect of feeding on protein synthesis. With the high vegetable-protein diet, protein breakdown in the absorptive state was not inhibited to the same extent as during the high-animal-protein diet, resulting in less net protein synthesis during the high-vegetable-protein diet than during the high-animal-protein diet. PMID- 9846852 TI - Plasma leptin influences gestational weight gain and postpartum weight retention. AB - BACKGROUND: Leptin, a product of the obese (ob) gene, is released from adipocytes. At the same body mass index, women have higher concentrations than men. Thus, during pregnancy, leptin may influence gestational weight gain and retention of a portion of that gain postpartum. OBJECTIVE: We examined the relation between plasma leptin at entry to prenatal care and subsequent changes in weight from entry to prenatal care (at 17 wk gestation, baseline) until 6 mo postpartum. DESIGN: This was an observational study of leptin, gestational weight gain, and postpartum weight retention (at 6 wk and 6 mo postpartum) in 103 low income pregnant women from Camden, NJ, with a pregravid body mass index (in kg/m2) in the normal range (19.8-26). RESULTS: After potential confounding variables were controlled for, leptin at entry significantly (P < 0.05) predicted weight gain in pregnancy, including measured rate of weight gain (x +/- SEE: 0.25 +/- 0.13 kg x unit log leptin(-1) x wk(-1)), measured rate of third-trimester weight gain (0.37 +/- 0.15 kg x unit log leptin(-1) x wk(-1)), rate of weight gain from recalled pregravid weight (0.23 +/- 0.09 kg x unit log leptin(-1) x wk( 1)), and net rate of gestational weight gain (0.22 +/- 0.09 kg x unit log leptin( 1) x wk(-1)). The leptin concentration at entry also significantly predicted retained weight in the postpartum period (at 6 mo: 7.29 +/- 3.33 kg/unit log leptin at entry) and marginally predicted changes in the sum of skinfold thicknesses (at 6 mo: 14.7 +/- 7.5 mm/unit log leptin at entry). CONCLUSION: These results suggest that a high leptin concentration at entry to prenatal care may predict an increased risk of overweight and obesity in vulnerable women. PMID- 9846853 TI - Serum transferrin receptor: a specific marker of iron deficiency in pregnancy. AB - BACKGROUND: Current markers of iron deficiency tend to be less reliable in pregnancy. OBJECTIVE: Our aim was to study the usefulness of soluble serum transferrin receptor (sTfR) as a marker for iron deficiency during early and late gestation and to define iron status in 254 pregnant Swedish women. DESIGN: We performed a cross-sectional and longitudinal evaluation of sTfR in comparison with concentrations of serum ferritin and hemoglobin in blood collected around gestational weeks 11 and 36. RESULTS: The specificity of sTfR was 100%. The sensitivity in relation to both anemia and depleted iron stores was approximately 70%, but this figure is less reliable because of few samples. sTfR in early pregnancy was low: 11% of women had a value below the reference interval. sTfR increased significantly from early to late pregnancy even in the group of women with persisting iron stores. In late pregnancy, 14% of women developed tissue iron deficiency and 5% had iron deficiency according to a combination of all 3 markers. CONCLUSIONS: sTfR seems to be a specific and sensitive marker of iron deficiency in pregnancy and may have advantages over serum ferritin and hemoglobin. The low sTfR concentration in early gestation seems to be caused by reduced erythropoiesis, whereas the increase from early to late pregnancy reflects increased erythropoiesis, and in case of iron deficiency, also tissue iron deficiency. Further studies are needed to verify whether decreased erythropoiesis reduces the possibility of detecting iron deficiency during early gestation by sTfR. PMID- 9846854 TI - Schistosoma mansoni infection and nutritional status in schoolchildren: a randomized, double-blind trial in northeastern Brazil. AB - Brazilian schoolchildren with mild- to moderate-intensity schistosome infections (<400 Schistosoma mansoni eggs/g stool) were randomly allocated to a treatment (oxamniquine) or placebo group in a double-blind fashion. Anthropometric measurements were made at baseline, 6 mo, and 1 y for 353 students. At baseline, the groups were not significantly different with respect to nutritional status or selected socioeconomic and biological characteristics, including anthropometric measures. One year later, significant differences were noted only in the nutritional status of boys treated for schistosome infection. Treated boys had greater measurements for weight, triceps skinfold thickness, midarm circumference, arm muscle area, and body mass index than untreated boys. They also showed significant increases over the year in weight, height, midarm circumference, and body mass index. The rates of improvement in weight and height were more accelerated in the first 6 mo after therapy than the last. These results indicate that, at least in boys, chronic S. mansoni infection at any intensity is detrimental to short-term growth and development. PMID- 9846855 TI - Randomized placebo-controlled clinical trial of the effect of a single high dose or daily low doses of vitamin A on the morbidity of hospitalized, malnourished children. AB - The effect of high-dose vitamin A supplementation on recovery from morbidity and on recovery from nosocomial morbidity of hospitalized children has been poorly studied and results are conflicting. The effect of daily, low doses has never been assessed. We investigated the effect of a single high dose and daily, low doses of vitamin A on diarrhea, acute lower respiratory tract infections (ALRIs), and all-cause fevers in 900 hospitalized preschool-age children in the Democratic Republic of Congo in a randomized, double-blind, placebo-controlled clinical trial. The high-dose treatment group received 200,000 IU vitamin A (100,000 IU if aged <12 mo) orally on the day of admission, the low-dose treatment group received 5000 IU vitamin A/d until discharge. Data on all-cause morbidity were collected daily. Mortality rates were not significantly different among the 3 groups. High-dose vitamin A supplementation had no significant effect on the duration of moderate or severe diarrhea nor on the duration and incidence of ALRIs and all-cause fevers. Children in the high-dose group with no edema had an increased risk of severe nosocomial diarrhea (relative risk: 2.42; 95% CI: 1.15, 5.11). Low-dose vitamin A supplementation significantly reduced the incidence of severe diarrhea in severely malnourished children (relative risk: 0.21; 95% CI: 0.07, 0.62) but showed no significant effect on the duration of moderate or severe diarrhea or on the duration and incidence of ALRIs and all-cause fevers. Supplementation with high doses of vitamin A did not reduce morbidity in this population of malnourished and subclinically vitamin A-deficient children; daily, low doses appeared more beneficial for severely malnourished children. PMID- 9846856 TI - Effect of zinc supplementation on growth and body composition of Ugandan preschool children: a randomized, controlled, intervention trial. AB - BACKGROUND: Despite ample food supplies, the incidence of childhood underweight and stunting remains high in Uganda. Many factors contribute to this situation, but the role of low zinc intakes has not been adequately explored. OBJECTIVE: Our objective was to study the effect of zinc supplementation on growth and body composition of preschool children by using the outcome measures of weight, height, and midupper arm circumference (MUAC). DESIGN: The study was randomized, double-blind, placebo-controlled, parallel, and 8 mo long, and incorporated 6 mo of zinc supplementation. Children (n = 153) aged 55.8 +/- 11.2 mo from 3 randomly selected nursery schools of medium, low, and very low socioeconomic status in a suburb of Kampala took part. The intervention comprised 10 mg Zn (as ZnSO4) or placebo daily in freshly prepared fruit juice, Monday to Friday inclusive. RESULTS: Zinc supplementation increased MUAC by the end of the study (P = 0.029) and led to greater weight gain in children from the school of medium socioeconomic status at 3 and 8 mo (P = 0.019 and P = 0.038, respectively). There was no effect on weight gain of the children from the other schools. Zinc supplementation had no influence on height. Infection rates (of which 82% were recorded as malaria) were lower in the zinc-supplemented group than in control subjects (P = 0.063). CONCLUSIONS: Zinc supplementation may counter the age related decrease in MUAC often observed in preschool children in developing countries. The study provides evidence that zinc may not be the most limiting nutrient for weight gain in children of poor nutritional status, but may become so as nutritional status improves. PMID- 9846857 TI - Inconsistencies in the findings of child nutrition surveys in Bangladesh. AB - BACKGROUND: The Bangladesh Bureau of Statistics (BBS) has been conducting nutrition surveys of young children at 2-3-y intervals since 1985. However, data on malnutrition determined on the basis of weight-for-age and height-for-age in different surveys are inconsistent. OBJECTIVE: The aim of this study was to investigate the reasons for inconsistencies in the findings of these surveys. DESIGN: Although the regular personnel of the BBS collected data on anthropometry and age in the previous surveys, 1995-1996 data were collected by locally recruited workers. Data on 100 children included in the 1995-1996 survey were also independently collected by BBS personnel. A comparison of the data from these 2 sources and the inconsistencies therein form the basis of this study. RESULTS: The average difference between the 2 sources (bias) was found to be approximately 1.8 mo for age and 0.13 cm for midupper arm circumference. Differences in weight and height were negligible; however, random error was found to be substantial in data on height. CONCLUSIONS: Bias in age was thought to be mainly responsible for the inconsistencies in the measures of malnutrition in the different child nutrition surveys in Bangladesh. A method for accurately collecting age data needs to be developed on the basis of a study in a demographic surveillance area where accurate age data are available. Until such a method is fully developed, midupper arm circumference should be routinely collected in all nutrition surveys. The importance of accuracy in weight and height data is emphasized. PMID- 9846858 TI - The nondigestible fat sucrose polyester does not stimulate gallbladder emptying in humans. AB - BACKGROUND: We determined the effect of oral ingestion of sucrose polyester, which was approved as a fat replacer in the United States, on gallbladder motility and on the release of cholecystokinin, the hormone that mediates gallbladder emptying. OBJECTIVE: Our objective was to measure effects of sucrose polyester on gallbladder emptying and cholecystokinin release. DESIGN: Eight healthy volunteers (3 men and 5 women) drank 60 mL sucrose polyester, digestible fat, or saline solution in a balanced crossover design on 3 separate days. RESULTS: Mean (+/-SEM) gallbladder emptying, when integrated over time, was low in response to both sucrose polyester (-150 +/- 214 mL x 120 min) and saline solution (-89 +/- 123 mL x 120 min). In contrast, there was marked emptying in response to digestible fat (1069 +/- 253 mL x 120 min). Sucrose polyester did not affect plasma cholecystokinin concentrations (-9.3 +/- 15.0 pmol x 120 min/L), whereas digestible fat resulted in a significant increase (89.5 +/- 44.8 pmol x 120 min/L, P = 0.014) compared with saline solution (-3.0 +/- 13.8 pmol x 120 min/L). CONCLUSIONS: Ingestion of sucrose polyester, in contrast with digestible fat, did not stimulate gallbladder emptying or release of cholecystokinin. PMID- 9846859 TI - Production rate of acetate during colonic fermentation of lactulose: a stable isotope study in humans. AB - BACKGROUND: Breath tests are currently used to qualitatively assess colonic fermentation; no quantitative estimations are available for healthy subjects. OBJECTIVE: This study describes a stable-isotope-dilution method to measure acetate production quantitatively from colonic bacterial fermentation. DESIGN: Six volunteers received a primed, constant, intravenous infusion of [1 13C]acetate at a rate of 1.01 +/- 0.04 micromol x kg(-1) x min(-1) for 7 h. They ingested 20 g pure lactulose after 1 h of the tracer infusion. Expired air and arterialized venous blood were sampled every 15 min. RESULTS: Before lactulose intake, the breath-hydrogen concentration was 7 +/- 2 ppm and the plasma acetate concentration and isotopic enrichment were 141 +/- 14 micromol/L and 14.8 +/- 1.4 moles percent excess, respectively. Whole-body acetate turnover was 6.0 +/- 0.7 micromol x kg(-1) x min(-1). After lactulose ingestion, maximum breath hydrogen and acetate concentrations reached 63 +/- 15 ppm (P = 0.004) and 313 +/- 25 micromol/L (P = 0.002), respectively, whereas [13C]acetate enrichment decreased to 9.9 +/- 1.3 moles percent excess (P = 0.03). Whole-body acetate turnover increased to 9.8 +/- 1.5 micromol x kg(-1) x min(-1) and later decreased almost to baseline values. Colonic lactulose fermentation yielded 140 +/- 12 mmol acetate over 6 h, representing 86% of the production based on stoichiometric equations. CONCLUSION: This new method provides a quantitative estimate of colonic carbohydrate fermentation via evaluation of acetate production. PMID- 9846860 TI - Changes in body composition and dietary intake induced by tumor necrosis factor alpha and corticosterone--individually and in combination. AB - Previous studies have shown that anorexia and reduced food intake are the main causes of weight loss in rats infused with tumor necrosis factor alpha (TNF alpha), with no influence on corticosterone concentrations. In contrast, in clinical sepsis, muscle wasting due to increased catabolism is associated with increased corticosteroid concentrations. We hypothesized that in the rat model, corticosterone potentiates the catabolic effect of TNF-alpha in amounts that by itself does not influence muscle catabolism. Orally fed rats were divided into 3 treatment groups: continuous infusion of TNF-alpha (TNF; 100 microg x kg(-1) x d( 1)), corticosterone (Cort; 50 microg x g(-1) x d(-1)), or both (TNF+Cort). Each group was compared with a respective pair-fed (PF) group. In addition an ad libitum (AL)-fed group receiving an infusion of physiologic saline was studied to observe unrestricted food intake and weight gain. After 4 d of infusion, dietary intake and weight gain were significantly higher in the Cort and AL groups than in the TNF and TNF+Cort groups. Although wet liver weights and protein contents were significantly higher in the Cort, TNF, and TNF+Cort groups than in their respective PF group, the TNF and TNF+Cort groups had lower relative carcass weights. The weight and protein content of the diaphragm were lower and nitrogen excretion was higher in the TNF+Cort group than in the respective PF group. The results suggest that TNF-alpha plus corticosterone had a specific catabolic effect on the diaphragm. In addition, together they increased overall nitrogen excretion. PMID- 9846861 TI - Age-related osteoporosis in Chinese: an evaluation of the response of intestinal calcium absorption and calcitropic hormones to dietary calcium deprivation. AB - BACKGROUND: Age-related osteoporosis may be associated with inefficient intestinal calcium absorption and bone remodeling. OBJECTIVE: We investigated the pathogenesis of age-related osteoporosis in Chinese women with habitual low calcium intakes. DESIGN: We studied the response of intestinal calcium absorption, calcitropic hormones, and biochemical bone markers to graded dietary calcium deprivation. RESULTS: The osteoporotic subjects (n = 25) had higher urinary calcium excretion (P < 0.05) and lower plasma 1,25-dihydroxyvitamin D concentrations (P < 0.02) than did age-matched control women (n = 25). Parathyroid hormone was not significantly different from that in age-matched control women but was significantly higher than in young women (n = 15, P < 0.05). Fractional 45Ca absorption was approximately 61% in all 3 groups when the diet was unmodified and increased to 71%, 69%, and 68% in the osteoporotic subjects, age-matched control women, and young women, respectively, when dietary calcium was reduced to 300 mg/d. When the osteoporotic women were calcium deprived, serum 1,25-dihydroxyvitamin D failed to increase but urinary calcium excretion persisted. In contrast, supplementation with 1200 mg Ca resulted in a lowering of parathyroid hormone (P < 0.005 compared with the unmodified diet) and 1,25-dihydroxyvitamin D (P < 0.01) and decreased fractional 45Ca absorption (P < 0.01), suggesting that the increased calcium intake was associated with a potent compensatory ability of the intestine and calcitropic hormones to adapt. Calcium supplementation lowered osteocalcin (P < 0.05) but not alkaline phosphatase, which remained elevated in the osteoporotic subjects at all stages. CONCLUSIONS: Elderly osteoporotic women had reduced 1,25-dihydroxyvitamin D production, excessive urinary calcium loss, and high bone turnover. The Chinese women had exceptionally potent intestinal calcium absorption. PMID- 9846862 TI - Dual-energy X-ray absorptiometry in the clinical evaluation of body composition and bone mineral density in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Depletion of fat-free mass (FFM) occurs in a considerable number of patients with chronic obstructive pulmonary disease (COPD). OBJECTIVE: The goal of the study was to determine whether dual-energy X-ray absorptiometry (DXA) is an applicable method in the clinical evaluation of body composition in COPD. DESIGN: In a cross-sectional study in 79 COPD patients participating in a pulmonary inpatient program and in 23 healthy volunteers, DXA was compared with deuterium dilution (Deu) in the estimation of FFM. Bone mineral density (BMD), a DXA measurement, was also compared between the 2 groups. RESULTS: FFM(DXA) was highly related to FFM(Deu) in men (R2 = 0.93, P < 0.001) and women (R2 = 0.91, P < 0.001). On average, DXA resulted in higher FFM values than did Deu in COPD patients (3.4 kg; P < 0.001) and in healthy volunteers (2.1 kg; P < 0.001). Furthermore, the intramethod difference in FFM was higher in men than in women in the COPD group (P < 0.05) and in healthy volunteers (P < 0.001). BMD was lower in the COPD group than in the healthy, age-matched volunteers (P < 0.001). In 56% of the COPD patients, there were indications of bone mineral loss, defined as a BMD <1 SD of a matched reference population provided by the software. BMD was <2 SDs in 36% of the COPD patients. CONCLUSIONS: DXA appears to be a suitable alternative method to Deu for assessing body composition and is also of value in identifying bone mineral loss in COPD patients, and is therefore applicable in the clinical evaluation of these patients. PMID- 9846863 TI - Effects of energy intake on the regulation of plasma leptin concentrations. PMID- 9846864 TI - Treatment of phenylketonuria. PMID- 9846865 TI - L-tryptophan in maternal phenylketonuria. PMID- 9846866 TI - Arise, go forth, and solve structures. PMID- 9846867 TI - Metastable states and folding free energy barriers. PMID- 9846868 TI - Location and function of linker histones. AB - The linker histones, H1 and its variant forms, have been implicated in the formation of higher orders of chromatin structure and gene repression. Three recent manuscripts have reexamined the location of H1 within the nucleosome particle and its function in the regulation of transcription. PMID- 9846869 TI - 100,000 protein structures for the biologist. AB - Structural genomics promises to deliver experimentally determined three dimensional structures for many thousands of protein domains. These domains will be carefully selected, so that the methods of fold assignment and comparative protein structure modeling will result in useful models for most other protein sequences. The impact on biology will be dramatic. PMID- 9846870 TI - Picture story. Fight the flu. PMID- 9846871 TI - HIV-1 A-rich RNA loop mimics the tRNA anticodon structure. AB - Interaction of HIV-1 genomic RNA and human tRNA(Lys)3 initiates viral reverse transcription. An adenosine-rich (A-rich) loop in HIV RNA mediates complex formation between tRNA and viral RNA. We have determined the structure of an A rich loop oligonucleotide using nuclear magnetic resonance spectroscopy. The loop structure is stabilized by a noncanonical G-A pair and a U-turn motif, which leads to stacking of the conserved adenosines. The structure has similarity to the tRNA anticodon structure, and suggests possible mechanisms for its role in initiation of reverse transcription. PMID- 9846872 TI - A minor groove RNA triple helix within the catalytic core of a group I intron. AB - Close packing of several double helical and single stranded RNA elements is required for the Tetrahymena group I ribozyme to achieve catalysis. The chemical basis of these packing interactions is largely unknown. Using nucleotide analog interference suppression (NAIS), we demonstrate that the P1 substrate helix and J8/7 single stranded segment form an extended minor groove triple helix within the catalytic core of the ribozyme. Because each triple in the complex is mediated by at least one 2'-OH group, this substrate recognition triplex is unique to RNA and is fundamentally different from major groove homopurine homopyrimidine triplexes. We have incorporated these biochemical data into a structural model of the ribozyme core that explains how the J8/7 strand organizes several helices within this complex RNA tertiary structure. PMID- 9846873 TI - Structure of a human DNA repair protein UBA domain that interacts with HIV-1 Vpr. AB - The HIV-1 protein Vpr is critical for a number of viral functions including a unique ability to arrest T-cells at a G2/M checkpoint and induce subsequent apoptosis. It has been shown to interact specifically with the second UBA (ubiquitin associated) domain found in the DNA repair protein HHR23A, a highly evolutionarily conserved protein. This domain is a commonly occurring sequence motif in some members of the ubiquitination pathway, UV excision repair proteins, and certain protein kinases. The three dimensional structure of the UBA domain, determined by NMR spectroscopy, is presented. The protein domain forms a compact three-helix bundle. One side of the protein has a hydrophobic surface that is the most likely Vpr target site. PMID- 9846874 TI - Structures of Cdc42 bound to the active and catalytically compromised forms of Cdc42GAP. AB - The Rho-related small GTP-binding protein Cdc42 has a low intrinsic GTPase activity that is significantly enhanced by its specific GTPase-activating protein, Cdc42GAP. In this report, we present the tertiary structure for the aluminum fluoride-promoted complex between Cdc42 and a catalytically active domain of Cdc42GAP as well as the complex between Cdc42 and the catalytically compromised Cdc42GAP(R305A) mutant. These structures, which mimic the transition state for the GTP hydrolytic reaction, show the presence of an AIF3 molecule, as was seen for the corresponding Ras-p120RasGAP complex, but in contrast to what has been reported for the Rho-Cdc42GAP complex or for heterotrimeric G protein alpha subunits, where AIF4- was observed. The Cdc42GAP stabilizes both the switch I and switch II domains of Cdc42 and contributes a highly conserved arginine (Arg 305) to the active site. Comparison of the structures for the wild type and mutant Cdc42GAP complexes provides important insights into the GAP-catalyzed GTP hydrolytic reaction. PMID- 9846875 TI - Iron center, substrate recognition and mechanism of peptide deformylase. AB - Eubacterial proteins are synthesized with a formyl group at the N-terminus which is hydrolytically removed from the nascent chain by the mononuclear iron enzyme peptide deformylase. Catalytic efficiency strongly depends on the identity of the bound metal. We have determined by X-ray crystallography the Fe2+, Ni2+ and Zn2+ forms of the Escherichia coli enzyme and a structure in complex with the reaction product Met-Ala-Ser. The structure of the complex, with the tripeptide bound at the active site, suggests detailed models for the mechanism of substrate recognition and catalysis. Differences of the protein structures due to the identity of the bound metal are extremely small and account only for the observation that Zn2+ binds more tightly than Fe2+ or Ni2+. The striking loss of catalytic activity of the Zn2+ form could be caused by its reluctance to change between tetrahedral and five-fold metal coordination believed to occur during catalysis. N-terminal formylation and subsequent deformylation PMID- 9846876 TI - MutY catalytic core, mutant and bound adenine structures define specificity for DNA repair enzyme superfamily. AB - The DNA glycosylase MutY, which is a member of the Helix-hairpin-Helix (HhH) DNA glycosylase superfamily, excises adenine from mispairs with 8-oxoguanine and guanine. High-resolution crystal structures of the MutY catalytic core (cMutY), the complex with bound adenine, and designed mutants reveal the basis for adenine specificity and glycosyl bond cleavage chemistry. The two cMutY helical domains form a positively-charged groove with the adenine-specific pocket at their interface. The Watson-Crick hydrogen bond partners of the bound adenine are substituted by protein atoms, confirming a nucleotide flipping mechanism, and supporting a specific DNA binding orientation by MutY and structurally related DNA glycosylases. PMID- 9846877 TI - Tunable alignment of macromolecules by filamentous phage yields dipolar coupling interactions. AB - Dipolar coupling interactions represent an extremely valuable source of long range distance and angle information that was previously not available for solution structure determinations of macromolecules. This is because observation of these dipolar coupling data requires creating an anisotropic environment for the macromolecule. Here we introduce a new method for generating tunable degrees of alignment of macromolecules by addition of magnetically aligned Pf1 filamentous bacteriophage as a cosolute. This phage-induced alignment technique has been used to study 1H-1H, 1H-13C, and 1H-15N dipolar coupling interactions in a DNA duplex, an RNA hairpin and several proteins including thioredoxin and apo calmodulin. The phage allow alignment of macromolecules over a wide range of temperature and solution conditions and thus represent a stable versatile method for generating partially aligned macromolecules in solution. PMID- 9846878 TI - Structure of a Numb PTB domain-peptide complex suggests a basis for diverse binding specificity. AB - The phosphotyrosine-binding (PTB) domain of Numb, a protein involved in asymmetric cell division, has recently been shown to bind to the adapter protein Lnx through an LDNPAY sequence, to the Numb-associated kinase (Nak) through a sequence that does not contain an NPXY motif and to GP(p)Y-containing peptides obtained from library screening. We show here that these diverse peptide sequences bind with comparable affinities to the Numb PTB domain at a common binding site on the surface of the protein. The NMR structure of the Numb PTB domain in complex with a GPpY-containing peptide reveals a novel mechanism of binding with the peptide in a helical turn that does not hydrogen bond to the PTB domain beta-sheet. These results suggest that PTB domains can potentially have multiple modes of peptide recognition and provide a structural basis from which the multiple functions of the Numb PTB domain during asymmetric cell division could arise. PMID- 9846879 TI - Crystal structure of a plant catechol oxidase containing a dicopper center. AB - Catechol oxidases are ubiquitous plant enzymes containing a dinuclear copper center. In the wound-response mechanism of the plant they catalyze the oxidation of a broad range of ortho-diphenols to the corresponding o-quinones coupled with the reduction of oxygen to water. The crystal structures of the enzyme from sweet potato in the resting dicupric Cu(II)-Cu(II) state, the reduced dicuprous Cu(I) Cu(I) form, and in complex with the inhibitor phenylthiourea were analyzed. The catalytic copper center is accommodated in a central four-helix-bundle located in a hydrophobic pocket close to the surface. Both metal binding sites are composed of three histidine ligands. His 109, ligated to the CuA site, is covalently linked to Cys 92 by an unusual thioether bond. Based on biochemical, spectroscopic and the presented structural data, a catalytical mechanism is proposed in which one of the oxygen atoms of the diphenolic substrate binds to CuB of the oxygenated enzyme. PMID- 9846880 TI - Structure of N-myristoyltransferase with bound myristoylCoA and peptide substrate analogs. AB - N-myristoyltransferase (Nmt) attaches myristate to the N-terminal glycine of many important eukaryotic and viral proteins. It is a target for anti-fungal and anti viral therapy. We have determined the structure, to 2.9 A resolution, of a ternary complex of Saccharomyces cerevisiae Nmt1p with bound myristoylCoA and peptide substrate analogs. The model reveals structural features that define the enzyme's substrate specificities and regulate the ordered binding and release of substrates and products. A novel catalytic mechanism is proposed involving deprotonation of the N-terminal ammonium of a peptide substrate by the enzyme's C terminal backbone carboxylate. PMID- 9846881 TI - Structure and mutagenesis of the Dbl homology domain. AB - Guanine nucleotide exchange factors in the Dbl family activate Rho GTPases by accelerating dissociation of bound GDP, promoting acquisition of the GTP-bound state. Dbl proteins possess a approximately 200 residue catalytic Dbl-homology (DH) domain, that is arranged in tandem with a C-terminal pleckstrin homology (PH) domain in nearly all cases. Here we report the solution structure of the DH domain of human PAK-interacting exchange protein (betaPIX). The domain is composed of 11 alpha-helices that form a flattened, elongated bundle. The structure explains a large body of mutagenesis data, which, along with sequence comparisons, identify the GTPase interaction site as a surface formed by three conserved helices near the center of one face of the domain. Proximity of the site to the DH C-terminus suggests a means by which PH-ligand interactions may be coupled to DH-GTPase interactions to regulate signaling through the Dbl proteins in vivo. PMID- 9846882 TI - Towards a new "AGE" of the chemistry and pathophysiology of the Maillard reaction. PMID- 9846883 TI - Cell activation by glycated proteins. AGE receptors, receptor recognition factors and functional classification of AGEs. AB - Proteins modified by advanced glycation endproducts (AGE) bind to cell surface receptors and other AGE binding proteins. AGE-binding receptors are: scavenger receptors types I and II, the receptor for advanced glycation endproducts (RAGE), oligosaccharyl transferase-48 (OST-48, AGE-R1), 80K-H phosphoprotein (AGE-R2) and galectin-3 (AGE-R3). AGE receptors are found in monocytes, macrophages, endothelial cells, pericytes, podocytes, astrocytes and microglia. AGE-modified proteins also bind to lysozyme and lactoferrin. A critical review of the evidence for receptors binding AGE-modified protein binding in vivo is presented. Scavenger receptors have only been shown to bind proteins modified by AGE to a much higher extent than found in vivo. 80K-H phosphoprotein is involved in FGFR3 signal transduction to MAP kinase, and may be involved in AGE-receptor signal transduction. Whether all of these proteins bind AGE-modified proteins in vivo is not yet clear. Cell activation in response to AGE-modified proteins is associated with increased expression of extracellular matrix proteins, vascular adhesion molecules, cytokines and growth factors. Depending on the cell type and concurrent signaling, this is associated with chemotaxis, angiogenesis, oxidative stress, cell proliferation or programmed cell death (PCD). Receptor recognition factors for agonism at the AGE receptor have been little studied but to date hydroimidazolones appear to be the most likely candidates. Pharmacologic inhibition of AGE receptor-mediated cell activation with specific antagonists may provide the basis for therapeutic intervention in diseases where AGE accumulation is a suspected etiological factor vascular complications of diabetes, macrovascular disease, renal insufficiency and Alzheimer's disease. PMID- 9846884 TI - Advanced glycation endproducts and cigarette smoking. AB - The incidence of certain ageing sequelae such as lung and cardiovascular disease and cataract are higher in smokers than in non-smokers. We recently proposed that certain components of mainstream cigarette smoke can react with plasma and extracellular matrix proteins to form covalent adducts with many of the properties of advanced glycation endproducts (AGE). AGEs have been implicated previously in the pathogenesis of the end-organ complications of diabetes and ageing, including cataract, atherosclerosis and renal insufficiency. In these circumstances, AGEs arise in vivo from the non-enzymatic reaction of reducing sugars with amino groups. Over time the initial Schiff base and Amadori products that form gradually undergo dehydration and rearrangement to produce reactive, carbonyl containing compounds with characteristic fluorescence and covalent crosslinking properties. Recent studies indicate that in smokers, tobacco-derived AGEs accumulate on plasma low density lipoprotein (LDL), structural proteins present within the vascular wall, and the lens proteins of the eye. These data point to a new and significant source of Maillard products in the human environment, significantly broaden the role of Maillard chemistry in pathological processes, and provide new insight into the pathogenesis of atherosclerosis and other diseases associated with tobacco usage. PMID- 9846885 TI - Impairment of endothelial function induced by glyc-oxidized lipoprotein a [Lp(a)]. AB - Diabetic patients develop endothelial dysfunction early in the course of the disease. Atherogenic lipoproteins such as LDL and Lp(a) are important risk factors for endothelial dysfunction and undergo nonenzymatic glycation in hyperglycaemia. Here we assessed whether glycation of Lp(a) potentiates its damaging influence on endothelial function. Human Lp(a) was glycated by dialyzation for 7 days against buffer containing 200 mmol/l glucose, or sham treated without glucose and oxidized by incubation with Cu++. The degree of glycation accounted to 32 +/- 4%, and glycation rendered Lp(a) more susceptible to oxidative modification when exposed to Cu++. Isolated rings of rabbit aorta were superfused with physiological salt solution, and isometric tension was recorded. Incubation of the aortic rings with sham-treated or with 30 microg/ml glycated Lp(a), not oxidized, had no influence on acetylcholine-induced, endothelium-dependent relaxation. Exposure of the aortic rings to 30 microg/ml oxidized non-glycated (ox) Lp(a) caused a significant inhibition (19% at 1 microM acetylcholine) of the endothelium-dependent relaxation. Incubation of aortic rings with 30 microg/ml oxidized glycated (glyc-ox) Lp(a) attenuated endothelium dependent relaxation more potently than oxLp(a) (by 34% at 1 microM acetylcholine). The presence of diethyl-dithio-carbamate (DDC), an inhibitor of the endogenous superoxide dismutase (SOD), potentiated the inhibition of relaxation induced by oxLp(a) and by glyc-oxLp(a) [38% inhibition at 1 microM acetylcholine for oxLp(a), and 49% inhibition at 1 microM acetylcholine for glyc oxLp(a)]. Co-incubation with the O2- scavenger 4,5-dihydroxy-1,3-benzene disulfonic acid disodium salt (TIRON) prevented the inhibition of relaxation by the oxidized lipoproteins, suggesting that enhanced NO-inactivation by O2- could be the underlying mechanism for the impairment of endothelium-dependent dilations by ox- and glyc-oxLp(a). The concentration of lysophosphatidycholine, a lipoprotein oxidation product and stimulus for O2- formation, was significantly enhanced in oxLp(a) and in glyc-oxLp(a) compared to native lipoproteins. CONCLUSION: Glycation enhances the endothelium-damaging influence of oxLp(a), presumably by enhancing oxidative stress. The likely mechanism for attenuation of endothelium-dependent dilations is increased formation of O2-, resulting in inactivation of nitric oxide. This mechanism may play an important role in diabetic patients and may contribute to disturbed organ perfusion. PMID- 9846886 TI - Cataract as a conformational disease--the Maillard reaction, alpha-crystallin and chemotherapy. AB - Cataract, the major cause of blindness world-wide, is associated with conformational changes and unfolding of proteins in the lens, which can arise directly as a result of post-translational modifications, induced by the Maillard reaction. In the lens, the stress protein alpha-crystallin, which is related to small heat-shock proteins and forms GroEL-like functional aggregates, can act as a chaperone-like protein to maintain transparency, sequestering unfolded protein, and inhibiting subsequent aggregation and insolubilisation. There are a number of criteria which enable the classification of cataract as a conformational disease, including not only the protein conformational change itself, resulting in aggregation and tissue deposition, but also the mechanisms for preventing such unfolding and aggregation. Post-translational modification of alphabeta crystallin results in loss of chaperone-like activity, and aspirin, ibuprofen and paracetamol can inhibit in vitro cross-linking events responsible for the loss of this activity. Of the many avenues available to block protein aggregation, common analgesics--and vitamin C--may provide a cost-effective route to explore further in the treatment of a range of conformational diseases. PMID- 9846887 TI - Advanced glycated albumin impairs protein degradation in the kidney proximal tubules cell line LLC-PK1. AB - Advanced glycation end-products (AGEs) are assumed to play a major role in the genesis of diabetic nephropathy and other diabetic complications. We studied the potential effect of AGEs on protein turnover and lysosomal proteinase activities in LLC-PK1 cells, a pig kidney proximal tubules cell line. Advanced glycated bovine serum albumin (AGE-BSA) was used as a model of AGEs and its action was compared to that of nonglycated BSA. AGE-BSA but not BSA (50 micromol/l) induced a significant increase in cell volume (BSA: 4870.6 +/- 74.8 fl, AGE-BSA: 5718.0 +/- 20.7 fl, p<0.01). Protein degradation rate was decreased by 13.8% after 48 hrs. incubation with AGE-BSA (p<0.01) while protein synthesis increased by 19,1%, (p<0.01). After incubation with AGE-BSA but not BSA activities of lysosomal cathepsins (B, L+B and H) decreased in a time- and dose-dependent fashion. This decline was neither caused by a shift in lysosomal pH outside the optimal range for cathepsins, nor by a direct inhibitory effect of AGEs modified proteins or peptides but most probably by inhibition of cathepsin B expression as measured by RT-PCR. It is supposed that impaired protease activities participated in decreased protein breakdown and cell enlargement. For the first time our data provide the evidence that AGEs induce hypertrophy of LLC-PK1 cells due to decreased protein breakdown resulting from reduced lysosomal proteinase activities with a concomitant stimulation of protein synthesis. PMID- 9846888 TI - Effects of continuous ambulatory peritoneal dialysis and kidney transplantation on advanced glycation endproducts in the skin and peritoneum. AB - Advanced glycation endproducts (AGE) form as a result of non-enzymatic reaction of reducing sugars with proteins. Patients with chronic renal failure (CRF) have elevated AGE in plasma, skin and peritoneum. We measured AGE in the skin and peritoneum of individuals with CRF, patients undergoing continuous ambulatory peritoneal dialysis (CAPD) and in renal transplant recipients (TR). Pentosidine concentration and collagen-linked fluorescence (CLF) were measured. Pentosidine and CLF correlated in all patient groups (CRF r=0.688, p<0.01; CAPD r=0.674, p<0.05; TR r=0.811, p<0.01). Successful kidney transplant reduced AGE levels in the skin (CRF 11.7 +/- 4.51 U/mg; TR 5.02 +/- 3.13 U/mg, p<0.00001) and peritoneum (CRF 17.5 +/- 6.16 U/mg, TR 9.4 +/- 4.97 U/mg, p<0.0001). However in contrast to the TR group, CLF in peritoneum increased following CAPD (CRF 17.5 +/ 6.16 U/mg; CAPD 24.2 +/- 10.4 U/mg; p=0.06). Our results suggest that AGE might be formed in the peritoneum during CAPD treatment. PMID- 9846889 TI - Factors in human serum interfere with the measurement of advanced glycation endproducts. AB - Advanced glycation endproducts (AGEs) have been implicated in the pathophysiology of coronary heart disease in ageing, diabetes and renal disease. Competitive enzyme-linked immunosorbent assays (ELISAs) have been developed to measure these compounds in serum, but as recognition of AGEs is both carrier protein- and antibody-dependent standardisation is problematic. We report here on another barrier to standardization, as yet unrecognised. During the development of an AGE ELISA, we found that serum samples did not dilute in parallel to AGE standards or each other. This finding was confirmed by recovery studies that showed over recovery of AGEs at high serum concentrations, but under-recovery at high dilutions of serum in assay buffer. We developed an inhibition assay to detect factors in serum capable of interacting directly with AGEs immobilised on microtitre plates. Binding of these factors prevented recognition of AGEs by a CML monoclonal antibody and a polyclonal anti-AGE antibody, and was neither sugar nor carrier protein-dependent. We detected the presence of this factor in all human sera tested and also in foetal calf serum. Pre-incubation of sera with AGEs or heat-treatment at 56 degrees C for 30 min. significantly reduced this binding. We are currently investigating the nature of this factor and the possibility that it may be complement. The effect of this factor on immunoassays for AGEs can only be detected by performing parallelism and recovery studies and we suggest the use of the method referred to in this paper to aid interpretation of parallelism data. PMID- 9846890 TI - The pathogenesis and consequences of AGE formation in uraemia and its treatment. AB - Advanced glycation endproducts (AGEs) accumulate in uraemia as a consequence of diminished clearance of low molecular weight forms which retain their reactivity and may subsequently combine with circulating and tissue macromolecules. Successful renal transplantation is the only form of renal replacement therapy which effectively clears these circulating AGEs; both haemodialysis and peritoneal dialysis are comparatively ineffective although high-flux haemodialysis confers some benefits. De novo AGE formation may be accelerated in uraemia due to carbonyl and oxidative stress leading to further accumulation. The consequences for the patient with chronic renal failure may be acceleration of vascular disease, renal failure progression and dialysis-related amyloidosis. Accelerated peritoneal AGE formation as a consequence of treatment with peritoneal dialysis fluids may be detrimental to peritoneal membrane function but does not appear to contribute to systemic elevation of AGEs. PMID- 9846891 TI - Hepatic advanced glycation endproduct binding is increased in experimental diabetes. AB - Advanced glycation endproducts (AGEs) have been implicated in the pathogenesis of diabetic complications. However, clearance pathways for these products have not been fully delineated. This study investigates changes in AGE binding in the liver in association with experimental diabetes using in vitro and in vivo radioautography techniques. Male Sprague-Dawley rats were randomised into control and diabetic rats and sacrificed after 3 weeks. Frozen liver sections (20 microm) were incubated with 125I-AGE-BSA. To further localise the AGE binding site, in vivo radioautography was performed by injection of 15 microCi of 125I-AGE-BSA into the abdominal aorta of the rat. Specific binding sites for AGEs were detected in the liver by in vitro radioautography. There was a significant increase in 125I-AGE binding in the liver of diabetic rats. Emulsion radioautography revealed that binding was localised primarily in Kupffer and liver endothelial cells. AGE binding sites were increased in the liver after 3 weeks of experimental diabetes. It remains speculative as to whether these binding sites represent AGE clearance receptors. PMID- 9846892 TI - Immunohistochemical detection of imidazolone and N(epsilon)-(carboxymethyl)lysine in aortas of hemodialysis patients. AB - The modification of long-lived proteins with advanced glycation endproducts (AGEs) has been hypothesised to contribute to the development of pathologies associated with uremia. Imidazolone and N(epsilon)-(carboxymethyl)lysine (CML) are common epitopes of AGE-modified proteins. Imidazolone is a reaction product of arginine with 3-deoxyglucosone (3-DG) which is markedly accumulated in uremic serum. CML is produced by glycoxidation, and represents a marker of oxidative stress. The specificity of anti-imidazolone antibody that we had developed was further examined using ELISA. The antibody reacted only with imidazolone derived from 3-DG and arginine, but did not react at all with the other imidazolone-like compounds such as reaction products of glyoxal, methylglyoxal, glucosone with arginine or a reaction product of 3-DG with creatine. Further, to determine if AGEs are involved in the development of atherosclerosis in hemodialysis (HD) patients, we studied the localisation of imidazolone and CML in the aortas obtained from HD patients by immunohistochemistry using the anti-imidazolone and anti-CML antibodies. Imidazolone and CML were localised in all atherosclerotic aortic walls of the HD patients. In conclusion, imidazolone and CML are localised in the characteristic lesions of atherosclerosis in HD patients. These results strongly suggest that imidazolone produced by 3-DG, and CML produced by glycoxidation may contribute to the development of atherosclerosis in uremic patients. PMID- 9846893 TI - Tau glycation is involved in aggregation of the protein but not in the formation of filaments. AB - A tau peptide, peptide 2R, with capacity for self assembly into filaments was used as a model to test the role of glycation on tau assembly or aggregation. Our results indicate that glycation of that peptide facilitates dimer formation but not assembly into filaments. However, glycation of tau results in the bundling of the tau filaments formed by glycosaminoglycan-induced polymerisation. These results suggest a role of glycation in the formation of covalent links among pre formed filaments but not in the assembly of those filaments. PMID- 9846894 TI - Modification of tau to an Alzheimer's type protein interferes with its interaction with microtubules. AB - The microtubule associated protein tau is the main structural component of paired helical filaments (PHFs), aberrant polymers found intracellularly in neurons of brains with the Alzheimer's disease. Glycation is one of the posttranslational modifications that has been found in tau from PHFs, but not in normal brain tau. Studies were carried out with purified tau protein subjected to chemical modifications, in order to further investigate the mechanisms of tau self association into PHFs. Tau was subjected to modifications affecting reactive lysyl residues, e.g., carbamoylation with potassium cyanate and glycation reaction with glucose. The effects of these modifications to produce functional alterations in tau capacity to bind brain tubulin and to induce microtubule assembly were investigated. Chemically-modified tau and tau of Alzheimer's type exhibited a similar microtubule interaction behavior as analysed by overlay assays, but those were different than normal tau controls. On the other hand, studies of the microtubule assembly kinetics indicated that the reported tau modifications resulted in a loss of its capacity to promote microtubule assembly from purified tubulin preparations. The data on the differences in the electrophoretic profiles, Western blots and the overlay patterns, along with those on the microtubule polymerisation of normal brain tau as compared with both modified and Alzheimer's tau, suggest changes in the functional behavior of this protein as a result of its structural modifications. These studies were complemented with an immunogold analysis at the electron microscope level, which indicated that the modified tau did not incorporate into assembled microtubules. These findings, combined with the results on tau chemical modifications suggest that the reactive lysine residues within functional domains on tau, e.g., those of the repetitive binding motifs, were affected by these modifications. Furthermore, these observations provide new clues to understand the anomalous interactions of tau in Alzheimer's disease. PMID- 9846895 TI - Advanced glycation endproducts (AGE) on IgG, a target for circulating antibodies in North American Indians with rheumatoid arthritis (RA). AB - Several tribes of North American Indians are known to have poor glucose control and are at a high risk of developing type 2 diabetes. Similarly some tribes also exhibit RA at a high frequency. We have recently determined that a subset of Caucasian patients with RA mount an immune response to IgG modified with advanced glycation endproducts (AGE). The AGE modifications on IgG in vivo include N(epsilon)-(carboxymethyl) lysine, imidazolone and pentosidine. The presence of IgG-AGE and the antibody response to the IgG-AGE in the Ojibwe tribe of First Nations native Indians where both NIDDM and RA are prevalent was investigated. AGE modified IgG and albumin were determined using a modified nitroblue tetrazolium assay. Rheumatoid factors (RFs) and IgM and IgA anti-IgG-AGE were detected by ELISA. Of the 108 individuals tested, 21 had RA only, 3 had both RA and type 2 diabetes, 30 had type 2 diabetes only and 51 had no diagnosed disease. AGE modified IgG was significantly elevated in the RA group compared to the diabetic group. IgM and IgA RFs were detected in 83% and 50% of the RA patients, compared to 31-37% and 7-10% of the diabetics or normal individuals. IgM anti-IgG AGE was detected in 54% of the RA patients, in contrast to 7-14% in the diabetics or normal individuals. IgA anti-IgG-AGE was detected in 42% of the RA patients and only 7 to 8% of the NIDDM or normal individuals. The IgM or IgA anti-IgG-AGE antibodies likely contribute to the accumulation of IgG-AGE, possibly through blocked clearance through AGE receptors. A trend towards more severe disease was seen in those Ojibwe RA patients with circulating anti-AGE antibodies. Non enzymatic glycation may be an important pathogenic link in the RA seen in North American Indians. PMID- 9846897 TI - Receptor for advanced glycation endproducts (RAGE) exhibits highly differential cellular and subcellular localisation in rat and human lung. AB - The transmembrane receptor (RAGE) of advanced glycation endproducts (AGEs), is abundantly present in the lung. Although the interaction of AGEs and RAGE plays an important role in vasculopathies, particularly in diabetes, the lung is not a classical target organ of diabetes. Thus, the role of RAGE in the lung is still obscure. This study sought to precisely localise RAGE in the lungs of rat and human by immunohistochemistry, double immunofluorescence and immunoelectron microscopy using a polyclonal antiserum developed against human recombinant RAGE. Anti-RAGE immunoreactivity was prominent in alveolar epithelial type I pneumocytes, while it was absent from type II pneumocytes and capillary endothelium. Cell type specificity was demonstrated by colocalisation with well established cell markers. Quantitative immunoelectron microscopy of cryo substituted, Lowicryl-embedded rat and human specimens demonstrated a unique labelling pattern of RAGE in that it selectively localised to the basal cell membrane of type I pneumocytes. Labelling pattern was independent of the mode of fixation. Equivalent labelling densities were calculated from a fibrotic rat lung 3 months after irradiation. This highly selective localisation of RAGE to the basal face of type I pneumocytes and its absence from capillary endothelium might explain the resistance of the lung to typical diabetic complications. PMID- 9846896 TI - Chemical modification of proteins by methylglyoxal. AB - Methylglyoxal is formed in vivo by spontaneous decomposition of triose phosphate intermediates in aerobic glycolysis. It may also be formed during oxidative degradation of both carbohydrates (pentoses and ascorbate) and lipids (arachidonate). In addition to reaction with arginine residues to form imidazolone adducts, methylglyoxal reacts with lysine residues in protein to form N(epsilon)-(carboxyethyl)lysine (CEL) and the imidazolium crosslink, methylglyoxal-lysine dimer (MOLD). Like the glycoxidation products, N(epsilon) (carboxymethyl)lysine (CML) and glyoxal-lysine dimer (GOLD) which are formed on reaction of glyoxal with protein, CEL and MOLD increase in lens proteins and skin collagen with age. CML and CEL also increase in skin collagen in diabetes, while all four compounds increase in plasma proteins in uremia. Overall, CML, CEL, GOLD and MOLD are quantitatively the major biomarkers of the Maillard reaction in tissue proteins. GOLD and MOLD, in particular, are present at 10-50 fold higher concentrations than the fluorescent crosslink, pentosidine. Together, these dicarbonyl-derived advanced glycation endproducts (AGEs) represent the major chemical modifications that accumulate in tissue proteins with age and in chronic diseases such as diabetes and atherosclerosis. PMID- 9846898 TI - Characterisation of a novel AGE-compound derived from lysine and 3 deoxyglucosone. AB - Dicarbonyl compounds are supposed to be reactive intermediates in the non enzymatic glycation of proteins. A process that eventually could lead to the development of late diabetic complications. Glyoxal lysine dimer (GOLD) and methylglyoxal lysine dimer (MOLD) have previously been described as such reactive dicarbonyls. Here a new compound 3-deoxyglucosone lysine dimer (DOLD), a cross link resulting from the reaction between hippuryl-lysine and 3-deoxyglucosone, has been isolated by HPLC and the structure determined by mass spectrometry and NMR. PMID- 9846899 TI - DNA-glycation leads to depurination by the loss of N2-carboxyethylguanine in vitro. AB - D-Glucose reacts with nucleosides, mainly guanosine, and DNA in a similar way as with proteins, and DNA-bound advanced glycation endproducts (AGE) are formed. Guanosine (Gua) was incubated with D-glucose or dihydroxyacetone (DHA) under variation of temperature and reaction time, and the formation of the glycation products N2-carboxyethylguanosine (CEG) and N2-carboxymethylguanosine (CMG) was monitored by HPLC/DAD. DHA proved to be a very potent glycating agent which specifically produces CEG. When deoxyguanosine (dGua) was glycated under the same conditions, formation of the glycation product N2-carboxyethyldeoxyguanosine (CEdG) resulted in a significant decrease in stability of the N-glycosidic bond. As a consequence more than 25% of CEdG was hydrolysed to give the corresponding guanine derivative CEguanine, whereas unmodified dGua was stable under these conditions. Glycated DNA was then incubated and depurination by the loss of CEguanine was monitored by HPLC. Significant release of CEguanine was observed, whereas free guanine was neither formed from glycated nor from control DNA in detectable amounts. These data suggest that CEdG which is formed during the glycation of DNA in vitro causes increased depurination of DNA and produces abasic sites. PMID- 9846900 TI - Fracture resistance of endodontically treated roots after restoration. AB - This study evaluated the shear strength resistance of endodontically treated roots that were restored by two different techniques. Twenty-seven recently extracted single-rooted teeth with similar anatomic characteristics were sectioned to obtain the same length for all specimens. Group I (GI) consisted of 14 roots restored with cast post-core (nickel-chromium alloy) and cemented using zinc phosphate cement; group II (GII) consisted of 13 roots restored with steel prefabricated posts (FKG) cemented with zinc phosphate cement and rotated with caution for anchorage. The crown portion of this group was made using a hybrid composite resin (Prisma APH). Results showed that on all specimens of GI the fracture occurred in the cervical root structure while in GII the composite resin fractured in all specimens. Statistical analysis showed a significantly higher resistance to fracture for GI than GII. The specimens were sectioned longitudinally for stereoscopic microscope analysis (63x) and did not show fracture lines in the dentin anchorage post area for any of the specimens. PMID- 9846901 TI - Artificial caries formation around fluoride-releasing restorations in roots. AB - Secondary caries is one of the most important factors leading to replacement of dental restorations. This investigation assessed the capacity of fluoride releasing restorative materials to resist caries in vitro when used in roots. Class 5 cavities were prepared in the buccal and lingual surfaces of 30 extracted premolars. The six materials used were: glass-ionomer cement (Fuji), glass ionomer cement with silver particles added (Ketac-silver), fluoride-containing composite resin (Tetric), composite resin (Silux plus), fluoride-containing amalgam (Fluor-Alloy) and high-copper amalgam (Dispersalloy). After 5 weeks in an acid gel for caries-like lesion formation, the teeth were sectioned longitudinally and examined with polarized light. The results showed that repair with glass-ionomer materials of a carious lesion may be of great importance in the prevention of secondary caries around the restorations in roots. PMID- 9846902 TI - Relationship of masticatory mandibular movements to masticatory performance of dentate adults: a method study. AB - This study evaluated a sieve method for measuring masticatory performance and determined the associations between masticatory performance and masticatory mandibular movements. Ten dentate adults and three complete denture-wearing subjects participated in the study. The masticatory performance indices for these subjects were determined after a 10 s masticatory sequence and another sequence that ended at the swallowing threshold. Almond was used as the test food and almond fragments were sized using a standard 0.65 mm square size brass wire sieve. Masticatory mandibular movements were simultaneously recorded by an optoelectronic device. The reproducibility of the masticatory performance tests was good (r = 0.98 and 0.82 for the 10 s and swallowing threshold test indices, respectively). The masticatory performance indices were markedly different between dentate and denture-wearing subjects. The 10 s masticatory performance index in the dentate subjects, showed moderate to rather strong correlation with all parameters of mandibular velocity (r = 0.6-0.7). The 10 s index also showed a strong negative correlation to the duration of the occlusal level phase and the total duration of the chewing cycle (r = -0.7 to -0.8) in the dentate subjects. This study confirms that masticatory performance levels are relatively stable and associated with the efficacy of specific masticatory mandibular movement parameters. PMID- 9846903 TI - Masticatory muscle responses associated with unloading of biting force during food crushing. AB - The aim of this study was to evaluate masticatory muscle responses during food crushing. Electromyograms were recorded simultaneously with mandibular movement and the sound of food fracture in eight healthy adult males at the superior and inferior heads of the lateral pterygoid muscles, the anterior belly of the digastric muscle, the temporal muscle, and the masseter muscle. After fracture of the food, silent periods of the elevator muscles and excitatory responses of the depressor muscles were observed. The latency and duration of the silent periods increased significantly with increasing incisal velocity after fracture. The mean latency of the excitatory responses of the depressor muscle was stable. After the application of anaesthetic into the temporomandibular joint capsule, the excitatory response disappeared, whereas the silent period was partly maintained. These findings suggest that the excitatory response of the depressor muscle occurs via a polysynaptic reflex arc, and that several reflex arcs are involved in the development of the silent period during food crushing. An important role for the temporomandibular joint receptors is implied. PMID- 9846904 TI - EMG activity of the orbicularis oris and mentalis muscles in children with malocclusion, incompetent lips and atypical swallowing--part I. AB - The function and the level of activity of the orbicularis oris (upper and lower), and of the mentalis muscles were verified electromyographically in resting position and in several movements and carried out in 18 children aged 8-12 years, divided into three groups: one with normal occlusion, and two with Class II division 1, with atypical swallowing and/or incompetent lips, who had received no orthodontic treatment. It was observed that, in a resting position with the lips separated, there was no activity in any of the muscles. When there was a contact of the lips, action potentials were recorded in the muscles studied, in those individuals with incompetent lips. In the movements of sucking either through a straw or a pacifier and the thumb, there were no differences among the groups. The incompetent lips group presented very marked muscle activity of the lower orbicularis oris and mentalis in the movement of sucking a lollipop. In deglutition of saliva, the orbicularis oris presented slight activity, and the mentalis, moderate. In deglutition of water, the mentalis presented very marked activity, whereas, the upper and lower orbicularis oris presented moderate and marked activities, respectively. PMID- 9846905 TI - Finite element analysis of non-axial versus axial loading of oral implants in the mandible of the dog. AB - The influence of axial and non-axial occlusal loads on the bone remodelling phenomena around oral implants in an animal experiment is simulated in a finite element analysis. The axial and non-axial loading conditions were introduced by inserting a bilaterally supported fixed partial prosthesis and a cantilever fixed partial prosthesis on two IMZ implants in the mandible of beagle dogs. Earlier quantitative and qualitative histological analyses revealed a statistically significant different remodelling response between both loading conditions. Two dimensional and three-dimensional models are built to analyse and compare von Mises equivalent stress, maximum principal stress, maximum principal strain and strain energy density distributions, first around a free-standing implant and subsequently around the implants of the two prosthesis designs under the respective resultant in vivo loads. Strong correlations between the calculated stress distributions in the surrounding bone tissue and the remodelling phenomena in the comparative animal model are observed. It is concluded that the highest bone remodelling events coincide with the regions of highest equivalent stress and that the major remodelling differences between axial and non-axial loading are largely determined by the horizontal stress component of the engendered stresses. PMID- 9846906 TI - Concomitant mandibular and head-neck movements during jaw opening-closing in man. AB - To test the hypothesis of a functional relationship between the human mandibular and cranio-cervical motor systems, head-neck movements during voluntary mandibular movements were studied in 10 healthy young adults, using a wireless optoelectronic system for three-dimensional (3D) movement recording. The subjects, unaware of the underlying aim of the study, were instructed to perform maximal jaw opening-closing tasks at fast and slow speed. Movements were quantified as 3D movement amplitudes. A consistent finding in all subjects was parallel and coordinated head-neck movements during both fast and slow jaw opening-closing tasks. Jaw opening was always accompanied by head-neck extension and jaw closing by head-neck flexion. Combined movement and electromyographic recordings showed concomitant neck muscle activity during head-neck movements, indicative of an active repositioning of the head. No differences in 3D movement amplitudes could be seen with respect to speed. The head movement was 50% of the mandibular movement during jaw opening, but significantly smaller (30-40%), during the jaw closing phase. In repeated tests, the 3D movement amplitudes of the concomitant head movements were less variable during slow jaw movement and during the jaw opening phase, than during fast and jaw closing movements, suggesting speed- and phase-related differences in the mechanisms controlling the integrated mandibular and head-neck motor acts. The present results give further support to the concept of a functional trigeminocervical coupling during jaw activities in man. PMID- 9846907 TI - Influence of finishing time on marginal sealing ability of new generation composite bonding systems. AB - Although variation in finishing techniques has been shown to affect microleakage, little research has been published on the influence of finishing time on the marginal sealing ability of new generation composite bonding systems. The objective of the present study was to evaluate the influence of finishing time on the enamel and dentine marginal sealing ability of four new generation composite systems. Two class V preparations, which were solely in enamel or dentine, were made on the buccal surfaces of 96 freshly extracted molar teeth. The teeth were randomly divided into four groups of 24 and restored with composite resin (Silux Plus) utilizing the following bonding systems: Scotchbond Multi-purpose, Fuji Bond LC, Prime & Bond 2.0 and Bisco One-step. Half of the restorations in each group were finished immediately after light polymerization and stored for 1 week. For the remaining restorations, finishing was delayed for 1 week. The storage medium was isotonic saline at 37 degrees C throughout the experiment. All restorations were then thermocycled, subjected to dye penetration testing, sectioned and scored. The results suggest that the finishing of composite restorations, bonded with the bonding systems evaluated, should be carried out immediately after light polymerization. Delayed finishing does not improve but instead can be detrimental to the marginal seal of the restorations. The effects of delayed finishing are, however, both bonding system and tissue specific. PMID- 9846908 TI - A 2-year follow-up of mandibular posture following an increase in occlusal vertical dimension beyond the clinical rest position with fixed restorations. AB - While resting mandibular posture is continuously changing, repeatable relations of 'physiologic rest position' (PRP) and 'clinical rest position' (CRP) are described in the literature. The PRP is defined as a position of minimal muscle activity and CRP as a more closed clinical reference relation. Relaxed resting posture (RRP) is a repeatable postural range obtained by operator-induced relaxation techniques. This article reports on measurements of mandibular posture in a patient group following an increase in occlusal vertical dimension (OVD) with fixed restorations beyond CRP over 2 years. The relationship of CRP and RRP in the restored and a non-restored control group is compared. The relationship of RRP and corresponding masseteric EMG values to baseline minimal EMG levels is reported for both groups. Resting face height adapted to the increased OVD and remained consistent over 1 and 2 years. The RRP was greater and significantly different from CRP in both groups (P = 0.0001). Results indicated that both CRP and RRP were postural ranges and not specific postural positions. The RRP occurred at minimum baseline EMG levels for both groups and appears to be consistent with physiologic rest position. The possibility of achieving minimal EMG levels at varying interocclusal rest space relations is discussed. PMID- 9846909 TI - Helicobacter pylori infection in children. AB - Helicobacter pylori colonizes the human stomach, especially during childhood. However, a variety of H. pylori strains exists, with major differences in virulence characteristics which probably account for different clinical symptoms, and the majority of infected subjects remains asymptomatic. Helicobacter pylori infection is correlated with socioeconomic conditions and hygienic circumstances, resulting in an extremely high prevalence in children in developing countries. Commercial screening tests are not capable of separating the more virulent strains (type I with vacuolating toxin VacA and CagA protein) from the less virulent strains (type II, VacA and CagA negative). Type I strains, but not type II, are associated with an increased risk for duodenal ulcer and gastric cancer. Therefore, future screening tests and vaccinations should focus on the type I strains. PMID- 9846910 TI - Folic acid in the treatment of acute watery diarrhoea in children: a double blind, randomized, controlled trial. AB - One-hundred and six male children aged 6-23 months with a history of acute watery diarrhoea of less than 72 h duration were randomized to receive either folic acid in a dose of 5 mg at 8-h intervals or placebo for 5 d. There were 54 children in the folic acid group and 52 in the placebo group. The admission characteristics were comparable between the two groups. No significant differences were observed in the intake of oral rehydration solution or stool output between the groups. The mean+/-SD of total stool output (g kg(-1)) was 532+/-476 vs 479+/-354 and the duration (h) of diarrhoea was 108+/-68 vs 103+/-53 in the folic acid vs placebo group, respectively. The findings, therefore, should have a positive influence on preventing the inappropriate use of folic acid in acute diarrhoea. PMID- 9846911 TI - Adenosine deaminase activity in protein-energy malnutrition. AB - Cell-mediated immunity (CMI) was evaluated in 69 children with protein-energy malnutrition (PEM) and 20 healthy controls. Significantly decreased responses to purified protein derivative (PPD) (p < 0.02) and absolute lymphocyte count (ALC) (p < 0.01) and increased serum adenosine deaminase (ADA) activity (p < 0.001) were observed in PEM cases compared with the controls. The mean values of ALC and ADA activity in PEM patients were 85.9% and 158.7% of the normal mean, respectively. A significant negative correlation was observed between the two parameters (r=- 0.2765, p < 0.01). The CMI tests were abnormal in all three grades of PEM, except for the response to PPD in grade I, when compared with the controls. No significant differences were found between infected and uninfected PEM cases. Thus, impaired CMI was observed not only in grades II and III but also in grade I PEM patients and the concomitant infection did not affect its status. However, ADA activity demonstrated a more pronounced change than the other tests. PMID- 9846912 TI - Effect of environmental and clinical factors on lung function and respiratory symptoms in adolescents with alpha1-antitrypsin deficiency. AB - Individuals identified in the Swedish neonatal alpha1-antitrypsin (AAT) screening study were followed prospectively from their first to their eighteenth year of life. The aim of this study was to analyse the effect of environmental factors, i.e. active and passive smoking, and of clinical factors on lung function and the occurrence of respiratory symptoms in AAT-deficient adolescents. The study group consisted of 88 protease inhibitor (Pi)ZZ and 40 PiSZ adolescents. Medical history including respiratory symptoms, and active and passive smoking were recorded at each follow-up up to the age of 18 y. Lung function tests were performed at the present check-up. At the age of 18 y, both forced expiratory volume in one second (FEV ) and FEV1/vital capacity (VC) were significantly lower in the smoking than in the non-smoking subgroup, and significantly more smokers than non-smokers reported the presence of phlegm. The mean FEV1/VC ratio was lower for those presently exposed to parental smoking. Multiple linear regression analysis indicated that clinical liver disease in early life, active smoking and parental smoking were independent determinants of FEV1/VC. The results suggest that marginal deviations in lung function and the symptom of phlegm among AAT deficient adolescents occur characteristically early in the subgroup of smokers. Parental smoking may contribute to decreased lung function. PMID- 9846913 TI - Expression of the VLA family of integrins in coeliac intestinal mucosa. AB - The integrins, a family of cell surface proteins, mediate cell adhesion and may influence within the intestinal mucosa processes such as migration and/or proliferation and differentiation of enterocytes and lymphocytes. The aim of this study was to examine the distribution pattern of integrin subunits (VLA alpha1, alpha2, alpha3, alpha4, alpha5, alpha6, beta1 chains) in normal intestinal mucosa and in that of patients with active coeliac disease (CD) and CD in remission. Immunohistochemical techniques and double immunostainings with monoclonal antibodies were used for investigation of the VLA alpha family of integrins and beta1 chain distribution. While the majority of the findings are consistent with the few data previously reported in the literature, surprising is the finding of a lack of expression of VLAalpha1 on the intraepithelial lymphocytes in the coeliac mucosa. The deficient VLA alpha1 expression on IEL in coeliac but not in normal mucosa may imply a genetic variation or a specific deficiency of gene expression during T cell differentiation and activation. PMID- 9846914 TI - Collagen-derived markers of bone metabolism in osteogenesis imperfecta. AB - Markers of bone formation [C-terminal and N-terminal propeptides of procollagen I (PICP, PINP), osteocalcin and alkaline phosphatase] and bone resorption [C terminal cross-linked telopeptide of collagen I (ICTP) and hydroxypyridinium cross-links, pyridinoline (Pyr) and deoxypyridinoline (Dpyr)] were measured in 78 osteogenesis imperfecta (OI) patients to investigate bone metabolism in vivo and relate marker concentrations to phenotype and in vitro collagen I defects, as shown by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). PICP and PINP were generally low, and the serum levels were lower in all children and adults with mild OI and a quantitative collagen defect than in patients with severe OI and a qualitative collagen I defect. ICTP, Pyr and Dpyr were generally normal or reduced, but elevated in severely affected adults with a qualitative collagen I defect. The in vivo findings correlated with in vitro results of collagen I SDS-PAGE. Bone turnover is reduced in OI children and mildly affected OI adults, whereas bone resorption is elevated in severely affected adults. These findings may prove helpful for diagnosis and decision-making regarding therapy in OI. PMID- 9846915 TI - Pelvic ultrasonography in normal girls and in girls with pubertal precocity. AB - This prospective study sought to evaluate the role of pelvic ultrasonography in differentiating between various types of pubertal precocity. A control group of 117 normal girls (aged 1.1-15.6) was studied and compared with 87 girls with premature sexual maturation (aged 1.1-9.2 y). Of these patients 19 had central precocious puberty (CPP), 48 had isolated premature thelarche (IPT) and 20 had premature adrenarche (IPA). Pelvic ultrasound variables evaluated were: (i) uterus: longitudinal diameter (uterine length), cross-sectional area (CSA) and fundo-cervical ratio; and (ii) ovaries: volume and morphology. Ovarian morphology was subdivided in 6 different appearances: solid, microcystic, paucicystic, multicystic, macrocystic, and major isolated cyst. In normal control girls, uterine length and CSA increased with age, although no cut-off values could be defined between different age ranges, and they were correlated with breast stage; fundo-cervical ratio was stable through childhood and increased after age 9. Ovarian volume was significantly greater in pubertal girls with breast stage 2 than in those with only pubic and/or axillary hair. There was a clear predominance of solid ovarian appearances in the age range 2-7, with the multicystic appearance being seen only after age 7, a minority being macrocystic. After age 10 all the different patterns were observed, and after age 13 the frequency of a macrocystic pattern increased. Significantly more mature ovarian appearances were observed in subjects with breast development compared with those without, independently of the presence of pubic hair. Patients with IPT had no significant differences in pelvic ultrasound measurements when compared with age matched controls. All the different morphological ovarian appearances were observed in IPT, in contrast to age-matched controls, where only the less mature patterns (solid, micro- and paucicystic) were seen. Patients with CPP had significantly more mature patterns of ovarian morphology compared with age matched controls, but did not differ from pubertal pre-menarcheal controls. Those patients with IPA differed from age-matched controls only in having significantly greater uterine length and CSA. Comparison of the pelvic ultrasound parameters between patient groups (IPT, CPP, IPA) and age-matched controls revealed significantly higher values in CPP for uterine length, uterine CSA and ovarian volume. Ovarian volume was also greater in IPT than in IPA. Ovarian morphology was significantly different in patients (IPT, CPP, IPA) compared with age-matched controls, but none of the ovarian morphological appearances was exclusive to a single condition. IN CONCLUSION: (i) pelvic ultrasound parameters increase progressively from birth to maturity, but no clear cut-off values can be established between age ranges; (ii) pelvic ultrasound variables reach adult values during puberty, with differences in the timing that may reflect geographical variations; (iii) the multicystic ovarian appearance occurs just before the onset of puberty; (iv) pelvic ultrasonography cannot always differentiate clearly between different disturbances of puberty and therefore cannot supersede other observations and investigations in the evaluation of pubertal disorders; and (v) in this study we propose a more detailed pelvic ultrasound terminology that can avoid apparent confusion in defining ovarian ultrasound appearance. PMID- 9846916 TI - Bone age in 116 untreated patients with Turner's syndrome rated by a computer assisted method (CASAS). AB - Bone age maturation in 116 untreated patients with Turner's syndrome was evaluated in a cross-sectional and longitudinal analysis. A total of 265 radiographs were rated using the TW2-RUS method on the computer-assisted skeletal age score (CASAS) system. Bone age was found to be retarded from the chronological age of 3 to 6y. Between the ages of 7 and 12 y bone age almost equalled chronological age and progressed normally at a rate of 1 y y(-1). Bone maturation slowed down thereafter and epiphyseal closure was not reached before the age of 17 y. Reference data are presented on bone age and a bone age maturation curve for untreated patients with Turner's syndrome to be used in clinical practice. In the assessment of bone age and bone age velocity in Turner's syndrome the CASAS system produced reliable and valid results. The absolute difference between repeated bone age ratings was 0.26 "y" (median) with a range of 0.00-0.56 "y". Future studies evaluating the effect of growth promoting treatment in Turner's syndrome should use a computerized method for the determination of bone age. PMID- 9846917 TI - Intensified treatment of acute childhood lymphoblastic leukaemia has improved prognosis, especially in non-high-risk patients: the Nordic experience of 2648 patients diagnosed between 1981 and 1996. Nordic Society of Paediatric Haematology and Oncology (NOPHO) AB - In a multinational, population-based study from the five Nordic countries (Denmark, Finland, Iceland, Norway and Sweden), 2648 children below 15 y of age were diagnosed with acute lymphoblastic leukaemia (ALL) in the years 1981-1996. The annual incidence was 3.9/100000 children and was stable throughout the study period. The development from regional or national protocols to common Nordic treatment protocols for all risk groups was completed in 1992 through a successive intensification of therapy, based on multidrug chemotherapy including pulses of methotrexate in high doses and avoidance of cranial irradiation in most children. For children with non-B-cell ALL (n=2602), the event-free survival (p EFS) increased from 0.53+/-0.02 (diagnosed 7/81-6/86) to 0.67+/-0.02 (7/86-12/91) to 0.78+/-0.02 (1/92-12/96). The corresponding p-EFS values at 5 y were 0.57, 0.70 and 0.78, respectively. The main improvements were seen in the group of children with non-high risk leukaemia, with 5-y p-EFS values increasing from 0.60 to 0.76 and 0.85 for the three periods. In high-risk patients, progress has been moderate, especially in children with high white blood cell values at diagnosis. During the last 5-y period, only 10% of the patients received cranial irradiation in first remission while 90% of the patients received high doses of cytostatic infusions (methotrexate isolated or combined with cytarabinoside) and multiple intrathecal injections of methotrexate as CNS-adjusted treatment without any indication of an increased CNS relapse rate. PMID- 9846918 TI - Bone mineral status in children with phenylketonuria under treatment. AB - Bone mineral status was assessed in 48 children with phenylketonuria (PKU) (20M, 28 F, aged 2.5-17 y). Bone density was measured in the distal third of the right forearm using single photon absorptiometry and was expressed as +/-SD with respect to age- and gender-matched controls. Serum calcium (Ca), magnesium (Mg), phosphorus (P), alkaline phosphatase (ALP), parathyroid hormone and 25 hydroxyvitamin D were measured in morning samples. The ratios of urinary Ca/creatinine (UCa/UCr), UP/UCr, UMg/UCr and hydroxyproline (OH-Pr)/UCr were calculated in urine samples collected over a period of 3 h. Patients' data were compared with those of 50 controls (22 M, 28 F, aged 3-15 y). The data showed severe osteopenia (below -2SD) in 22/48 patients. Bone loss was more prominent in patients over 8 y old. Bone density correlated significantly with age (r=-0.56,p < 0.001) and with Phe (r=-0.49, p < 0.007) but did not correlate with the other biochemical indices studied. Comparing PKU children with controls, significantly higher serum calcium and magnesium (p=0.04, p < 0.001, respectively), lower ALP (p=0.01), higher UCa/UCr ratio (p < 0.001), lower UP/UCr (p < 0.001) and lower UOH-Pr/UCr (p < 0.001) were found. Dietary compliance was poor in patients over the age of 8y, as only 3/22 of < or = 8y had mean serum phenylalanine >10mgdl( 1), in contrast to 21/26 in the older group. It is clear from the data that osteopenia is commonly found in PKU patients from early life. The biochemical data indicate a metabolic state of low bone turnover in PKU patients. In conclusion, a better, more restricted diet may correct osteopenia. PMID- 9846919 TI - Lack of precision in neonatal death classifications based on the underlying causes of death stated on death certificates. AB - Large-scale analyses of causes of neonatal deaths are usually based on death certificate information. A new computer-based method has been introduced to define the cause of stillbirths and neonatal deaths in large amounts of material and to classify them according to two different models [Wigglesworth and Neonatal and Intrauterine death Classification according to (a)Etiology (NICE)]. The method is based on a combination of detailed information from health care registries and the death-certificate information. The present study aimed to compare these two classification models with a previously published method based solely on death certificate information [International Collaborative Effort (ICE)]. The study population comprised 2378 neonatal deaths in Sweden between 1987 and 1992. Cross-tabulation was made between the ICE classification and the other two classification models. In addition, case examples are presented in detail, exemplifying how classification errors arose. The ICE classification gives a rather low precision, notably for two important causes of death: asphyxia and immaturity. Among 328 infants dying from asphyxia according to computerized Wigglesworth classification, ICE classified 59% as asphyxia and 22% were labelled immaturity. When ICE classified the deaths as due to asphyxia, this was verified in only 50%. Among 792 infants dying from immaturity according to computerized Wigglesworth classification, 64% were classified as such by ICE. The findings cast doubts on the results of studies based exclusively on death-certificate information. Whenever possible in the analysis of neonatal deaths, death certificate information should be supplemented with more detailed data. The computer-based method introduced here makes such analyses possible for large databases. PMID- 9846920 TI - Screening for visual and ocular disorders in children, evaluation of the system in Sweden. AB - The aim of this study was to evaluate the visual screening system in Sweden. We have made a retrospective investigation of the results of screening for ocular disease and visual impairment of all children born in 1982 in three Swedish communities. The records from screening examinations from 0 to 10 y and from diagnostic follow-up at the departments of ophthalmology that the children were referred to were inspected. The data were used to evaluate the efficiency of the Swedish visual screening system. The study included 3126 children. The attendance rate at the 4-y examination was better than 99%. The sensitivity of the 4- and 5.5-y screening examinations was on the average 92% and the specificity was 97%. The average number of false negative cases at 4 y was 5.6 in 1000 (0.56%). With this screening and subsequent diagnosis and treatment, the prevalence of amblyopia at different levels of visual acuity at the age of 10 y was: 0.06% with visual acuity < or = 0.1, 0.9% with visual acuity < or = 0.5 and 1.7% with visual acuity < or = 0.7. In spite of largely unchanged pressure of amblyogenic factors in the population, the prevalence of deep and moderate amblyopia has been markedly reduced by screening and early treatment. PMID- 9846921 TI - Low-dose doxapram for treatment of apnoea following early weaning in very low birthweight infants: a randomized, double-blind study. AB - The effects of low-dose doxapram (0.5 mg kg(-1)h(-1)) in combination with caffeine were evaluated on apnoea frequency following weaning from mechanical ventilation, and on blood pressure, in very low birthweight (BW) premature infants. Twenty-nine infants with BW < or=1250 g, gestational age at birth (GA) <34 weeks and postnatal age <5 d, who required minimal respiratory support, were included. Following randomization, they received a loading dose of caffeine citrate and a continuous infusion of doxapram (doxapram, n=14) or placebo (n=15) was started. They were extubated 8 h after starting the infusion, which was continued for 5 d. During this period, weaning was well tolerated in both groups, apnoeas occurred less frequently and there was a greater increase in systolic blood pressure in infants treated with doxapram than in controls. Plasma doxapram levels were also higher than expected. It is therefore suggested that doxapram, even at low doses, should not be used during the first few days of life. Careful monitoring of blood pressure is required if doxapram is used later. PMID- 9846922 TI - Transepidermal water loss in infants born at 24 and 25 weeks of gestation. AB - The rate of evaporation of water from the skin of 13 infants born at 24 (n=3) and 25 (n=10) weeks of gestation was measured on the first day after birth and at postnatal ages of 1, 3, 7 and 28 d, using the gradient method. Transepidermal water loss was estimated from this rate and corrected to an ambient relative humidity (RH) of 50%. Transepidermal water loss, corrected to 50% RH, was high on the first day after birth (58.4+/-14.8 g m(-2) h(-1)) and remained at the same level during the second day (59.3+/-17.6 g m(-2)h(-1)). It then decreased significantly to 43.8+/-9.5 at a postnatal age of 3 d, 36.1+/-12.6 at 7 d and 24.2+/-7.7 g m(-2) h(-1) at 28 d (p < 0.001). Within the group investigated, there was no significant correlation between transepidermal water loss and body or skin temperature, birth weight, gender, mode of delivery or gestational age. Transepidermal water loss on the first day after birth was somewhat lower than the highest losses previously found in infants born at 25 weeks of gestation, and of the same magnitude as previously reported for infants born at 25-27 weeks. Transepidermal water loss at postnatal ages of 1, 3, 7 and 28 d in the present study was higher than that previously found in the group of infants born at 25-27 weeks. In conclusion, in infants born at 24-25 completed weeks of gestation transepidermal water loss was high immediately after birth and decreased with increasing postnatal age, but at a slower rate than previously reported for slightly more mature infants. PMID- 9846923 TI - Methaemoglobin formation after the use of EMLA cream in term neonates. AB - EMLA cream 5% (a eutectic mixture of lidocaine and prilocaine) is a topical anaesthetic that has become widely used to minimize pain from venipuncture in children. It has not, however, been recommended in neonates owing to the potential risk of methaemoglobinaemia induced by prilocaine. The aim of this study was to establish the safety of 1 g EMLA cream 5% used on intact skin in term neonates. Forty-seven neonates, aged 0-3 months, with a postconceptual age of > or = 37 weeks and a body weight between 2.8 and 5.7 kg, were included in a double-blind, randomized, placebo-controlled study. After baseline observations a total dose of 1.0 g EMLA/placebo was applied to two sites (0.5 g site(-1)) for 60 70 min. Venous methaemoglobin (metHb) levels were determined in each patient at baseline and at three randomly assigned times, 0.5-18 h after application. Following application of the cream, the mean metHb levels were 1.17% (range 0.50 2.53) in the EMLA group and 0.96% (range 0.50-1.53) in the placebo group. The metHb concentrations were significantly higher in the EMLA group in the intervals from 3.5 to 13 h after application than in the placebo group, but were well below potentially harmful levels. Based on these results, a 1-h application of 1 g EMLA cream is safe when used on the intact skin of term neonates below 3 months of age. PMID- 9846924 TI - Coma in a premature infant associated with the transdermal absorption of propylene glycol. AB - A case of propylene glycol intoxication in a premature infant is reported. The infant went into a state of coma after treatment for burns with antiseptic dressings. Cessation of the topical treatment resulted in complete recovery. An exceptionally high level of the dressings' solvent, propylene glycol, found in the urinary chromatogram, was believed to be the causative agent. It is suggested that topical preparations containing propylene glycol should not be used in premature infants during the first weeks of life. PMID- 9846925 TI - Is homozygous alpha-thalassaemia a lethal condition in the 1990s? AB - Two cases of homozygous alpha-thalassaemia who received active treatment in accordance with parental wishes are reported. One infant survived and the other, although successfully weaned off mechanical respiratory support, unexpectedly developed portal vein thrombosis and died. Homozygous alpha-thalassaemia, a condition previously considered to be universally fatal, and an indication for therapeutic abortion, is now potentially curable with advances in diagnostic technology and treatment. However, active management of these cases raises serious ethical questions and has major financial implications on the health-care system. Invasive prenatal and intensive postnatal interventions should remain experimental and cannot be recommended as routine clinical practice until the questions of long-term neurodevelopmental outcome, and the morbidity and mortality associated with bone-marrow transplantation have been fully addressed. As a result of advances in information technology, more and more parents of affected foetuses are likely to request active treatment. PMID- 9846926 TI - Mediastinal lymphadenopathy: a variant of incomplete Kawasaki disease. AB - A 14-month-old girl presented with a 4-d history of fever and generalized exanthema. Four characteristic symptoms of incomplete Kawasaki disease (KD) were present on admission (fever, rash, non-purulent conjunctival injection, oropharyngeal changes) and then followed by oedema of the hands and feet and mild plantar desquamation. The typical laboratory features of KD, such as elevated erythrocyte sedimentation rate, leukocytosis, thrombocytosis, and positive C reactive protein were also seen. Ultrasound examination of the mediastinum revealed the presence of a lymph node, 30 mm in diameter, below the tracheal carina. Thoracic CT scan confirmed the mediastinal lymph node. The patient was treated with aspirin and intravenous gamma-globulin. Ultrasound study of the mediastinum, which was carried out 6 weeks after hospital discharge, showed that the lymph node had disappeared. This case illustrates that lymph nodes other than cervical lymphadenopathy should be sought when the diagnosis of classical or atypical KD is suspected. PMID- 9846927 TI - Gastrostomy button causing perforation of the posterior gastric wall. AB - A gastrostomy button complication, not previously reported, is described. The button, with an inflatable balloon, was used for nutrition. The patient had had a gastrostomy for 4.5 y, with the same type of gastrostomy button for the previous 2 y and the same device for 1 y. The tip of the button caused a perforation of the posterior stomach wall, leading to death. PMID- 9846928 TI - Transient trimethylaminuria in childhood. AB - Trimethylaminuria, also called fish-odour syndrome, is an inherited disorder caused by deficient N-oxidation and increased excretion of trimethylamine. This study reports on two unrelated and otherwise healthy children with transient trimethylaminuria, a hitherto unknown abnormality, without N-oxidation deficiency. This demonstrates that a diagnosis of fish-odour syndrome should include the analysis of urinary excretion not only of trimethylamine but also of trimethylamine-N-oxide. Since transient trimethylaminuria may be a common condition and no cause for it can presently be recognized, such patients will require careful follow-up. PMID- 9846929 TI - Zidovudine treatment prevents vertical transmission of human immunodeficiency virus-1 independently of viral load. PMID- 9846930 TI - The Microcirculation Physiome Project. AB - Presented is a discussion of steps towards the creation of a database of the microcirculation encompassing anatomical and functional experimental data, and conceptual and computational models. The discussion includes issues of database utility, organization, data deposition, and linkage to other databases. The database will span levels from gene to tissue and will serve both research and educational purposes. PMID- 9846931 TI - Accounting for the heterogeneity of capillary transit times in modeling multiple indicator dilution data. AB - To mathematically model multiple indicator dilution (MID) data for the purpose of estimating parameters descriptive of indicator-tissue interactions, it is necessary to account for the effects of the distribution of capillary transit times, h(c)(t). In this paper, we present an efficient approach for incorporating h(c)(t) in the mathematical modeling of MID data. In this method, the solution of the model partial differential equations obtained at different locations along the model capillary having the longest transit time provides the outflow concentrations for all capillaries. When weighted by h(c)(t), these capillary outflow concentrations provide the outflow concentration versus time curve for the capillary bed. The method is appropriate whether the available data on capillary dispersion are in terms of capillary transit time or relative flow distributions, and whether the dispersion results from convection time differences among heterogeneous parallel pathways or axial diffusion along individual pathways. Finally, we show that the knowledge of a relationship among the moments of h(c)(t), rather than h(c)(t) per se, is sufficient information to account for the effect of h(c)(t) in the mathematical modeling interpretation of MID data. This relationship can be determined by including a flow-limited indicator in the injected bolus, thus providing an efficient means for obtaining the experimental data sufficient to account for capillary flow and transit time heterogeneity in MID modeling. PMID- 9846933 TI - Computational analysis of flow in a curved tube model of the coronary arteries: effects of time-varying curvature. AB - The flow through a curved tube whose radius of curvature varies with time was studied in order to better understand flow patterns in coronary arteries. A computational flow model was constructed using commercially available software. The artery model featured a uniform circular cross section, and the curvature was assumed to be constant along the tube, and in one plane. The computational model was verified with the use of a dynamically similar in vitro apparatus. A steady uniform velocity was prescribed at the entrance at a Reynolds number of 300. Two sets of results were obtained: one in which the curvature was held constant at the mean, maximum and minimum radii of curvature (quasistatic), and another in which the curvature was varied sinusoidally in time at a frequency of I Hz (dynamic). The results of the dynamic analysis showed that the wall shear rates varied as much as 52% of the static mean wall shear rate within a region of 10 tube diameters from the inlet. The results of the dynamic analysis were within 6% of the quasistatic predictions. Realistic modeling of the deforming geometry is important in determining which locations in the coronary arteries are subjected to low and oscillating wall shear stresses, flow patterns that have been associated with atherogenesis. PMID- 9846932 TI - Influence of gravity on cardiac performance. AB - Results obtained by the investigators in ground-based experiments and in two parabolic flight series of tests aboard the NASA KC-135 aircraft with a hydraulic simulator of the human systemic circulation have confirmed that a simple lack of hydrostatic pressure within an artificial ventricle causes a decrease in stroke volume of 20%-50%. A corresponding drop in stroke volume (SV) and cardiac output (CO) was observed over a range of atrial pressures (AP), representing a rightward shift of the classic CO versus AP cardiac function curve. These results are in agreement with echocardiographic experiments performed on space shuttle flights, where an average decrease in SV of 15% was measured following a three-day period of adaptation to weightlessness. The similarity of behavior of the hydraulic model to the human system suggests that the simple physical effects of the lack of hydrostatic pressure may be an important mechanism for the observed changes in cardiac performance in astronauts during the weightlessness of space flight. PMID- 9846934 TI - Effects of radial wall motion and flow waveform on the wall shear rate distribution in the divergent vascular graft. AB - Among the hemodynamic factors influencing intimal hyperplasia in the anastomotic region of a vascular graft, wall shear rate is believed to be one of the most important. We would like to study the effects radial wall motion on the wall shear rate distribution in the end-to-end anastomosis model of an artery and a divergent graft. Rigid and elastic models are constructed and the wall shear rate distributions are measured along the anastomosis using photochromic flow visualization method for carotid and femoral flow waveform. The mean and peak of shear rate decrease along the divergent graft, and the decreases are more significant in the elastic model. The shear rate waves are decomposed using the Fourier transform in order to separate the effects of radial wall motion and geometry. The percentage reductions of mean wall shear rates compared to steady shear rates at mean flow are calculated, and additional 8% (carotid) and 22% (femoral) reductions are observed in the elastic models near the end of the divergent graft. Also radial wall motion decreases the amplitudes of higher harmonics of wall shear rates in the elastic models. Since radial wall motion may affect the flow field differently for different geometry, wall elasticity should be considered in studying arterial hemodynamics. PMID- 9846935 TI - Noninvasive measurement of the human brachial artery pressure-area relation in collapse and hypertension. AB - A noninvasive method to obtain pressure-lumen area (P-A) measurements of the human brachial artery is introduced. The data obtained from this method are analyzed using a mathematical model of the relationship between vessel pressure and lumen area including vessel collapse and hypertension. An occlusive arm cuff is applied to the brachial artery of ten normal subjects. The cuff compliance is determined continuously by means of a known external volume calibration pump. This permits the computation of the P-A curve of the brachial artery under the cuff. A model is applied to analyze the P-A relation of each subject. The results show that the lumen area varies considerably between subjects. The in vivo resting P-A curve of the brachial artery possesses features similar to that of in vitro measurements. A primary difference is that the buckling pressure is higher in vivo, presumably due to axial tension, as opposed to in vitro where it is near zero or negative. It is found that hypertension causes a shift in the P-A curve towards larger lumen areas. Also, the compliance-pressure curve is shown to shift towards higher transmural pressures. Increased lumen area provides an adaptive mechanism by which compliance can be maintained constant in the face of elevated blood pressure, in spite of diminished distensibility. PMID- 9846936 TI - Finite element modeling of three-dimensional pulsatile flow in the abdominal aorta: relevance to atherosclerosis. AB - The infrarenal abdominal aorta is particularly prone to atherosclerotic plaque formation while the thoracic aorta is relatively resistant. Localized differences in hemodynamic conditions, including differences in velocity profiles, wall shear stress, and recirculation zones have been implicated in the differential localization of disease in the infrarenal aorta. A comprehensive computational framework was developed, utilizing a stabilized, time accurate, finite element method, to solve the equations governing blood flow in a model of a normal human abdominal aorta under simulated rest, pulsatile, flow conditions. Flow patterns and wall shear stress were computed. A recirculation zone was observed to form along the posterior wall of the infrarenal aorta. Low time-averaged wall shear stress and high shear stress temporal oscillations, as measured by an oscillatory shear index, were present in this location, along the posterior wall opposite the superior mesenteric artery and along the anterior wall between the superior and inferior mesenteric arteries. These regions were noted to coincide with a high probability-of-occurrence of sudanophilic lesions as reported by Cornhill et al. (Monogr. Atheroscler. 15:13-19, 1990). This numerical investigation provides detailed quantitative data on hemodynamic conditions in the abdominal aorta heretofore lacking in the study of the localization of atherosclerotic disease. PMID- 9846937 TI - Dynamic thermography: analysis of hand temperature during exercise. AB - Exercise has a noted effect on skin blood flow and temperature. We aimed to characterize the normal skin temperature response to exercise by thermographic imaging. A study was conducted on ten healthy and active subjects (age=25.8+/-0.7 years) who were exposed to graded exercise for determination of maximal oxygen consumption (VO2 max), and subsequently to constant loads corresponding to 50%, 70%, and 90% of VO2 max. The skin temperature response during 20 min of constant load exercise is characterized by an initial descending limb, an ascending limb and a quasi-steady-state period. For 50% VO2 max, the temperature decrease rate was - 0.0075+/-0.001 degrees C/s during a time interval of 390+/-47 s and the temperature increase rate was 0.0055+/-0.0031 degrees C/s during a time interval of 484+/-99 s. The level of load did not influence the temperature decrease and increase rates. In contrast, during graded load exercise, a continuous temperature decrease of -0.0049+/-0.0032 degrees C/s was observed throughout the test. In summary, the thermographic skin response to exercise is characterized by a specific pattern which reflects the dynamic balance between hemodynamic and thermoregulatory processes. PMID- 9846938 TI - Endocardial potential mapping from a noncontact nonexpandable catheter: a feasibility study. AB - In previous studies, we established methodology for reconstructing endocardial potentials, electrograms and isochrones from a non-contact intracavitary probe during a single beat. The probe was too large to be introduced percutaneously. Here we examine the possibility of similar mapping with a small multielectrode catheter that could be introduced percutaneously and does not expand inside the cavity. Cavity geometry and endocardial potentials were recorded in an isolated canine left ventricle. Simulated catheter probes were introduced into the cavity. Probe potentials were computed from the measured endocardial potentials and perturbed to include measurement noise, geometrical errors, and limited electrode density. Endocardial potentials were then reconstructed from the perturbed probe potentials and compared to the actual measured potentials. Of all probes simulated, a 3.0 mm (9F) catheter that assumes a curved geometry (e.g., a J shape) inside the cavity performed best (better than a larger 7.6 mm cylinder simulating an inflatable probe). Without bending, a straight cylindrical probe of the same size (9F, 3.0 mm) did not perform well. Sixty probe electrodes were needed for accurate reconstruction. The J-probe reconstruction was very robust in the presence of noise (10%) and of geometry errors (3 mm shift, 10 degrees rotation). The results demonstrate the feasibility of accurate single-beat endocardial mapping using a 9F percutaneous multielectrode catheter that assumes a J shape in the cavity without the need for expansion (e.g., into a balloon or a "basket"). The robustness of the procedure to noise and geometrical errors suggests its applicability in the clinical EP laboratory and the possibility of determining probe position in vivo using current imaging modalities. PMID- 9846939 TI - Quantitative characterization of epicardial wave fronts during regional ischemia and elevated extracellular potassium ion concentration. AB - This study applied zero-delay wave number spectral estimation as a means of quantifying the changes in activation and recovery sequences of propagating plane waves on the epicardial surface of in situ porcine hearts during regional hyperkalemia and ischemia. Unipolar electrograms (104) were recorded from the left ventricular surface of nine hearts using a plaque electrode array with 1 mm spatial sampling intervals. The objectives were (1) to define a set of parameters capable of quantifying the spatial and temporal changes in measured extracellular potentials associated with localized ischemia prior to the onset of conduction block; (2) to elevate regional levels of extracellular potassium ion concentration and quantify potential changes due to this known physiologic manipulation; and (3) to use quantitative parameters to make statistical comparisons in order to distinguish wave fronts during normal, ischemic and hyperkalemic conditions. Results showed that the parameters of wave number and average temporal frequency and the associated power, as determined from the wave number spectrum, provided statistically significant (p<0.05) quantification of changes in wave front features during normal and ischemic or hyperkalemic conditions. The results were consistent with results obtained from conventional time-space domain methods like isochronal mapping and electrograms, with the advantage of a quantitative result enabling simple comparisons and trend analysis for large numbers of heart beats. PMID- 9846940 TI - Simulated epicardial potential maps during paced activation reflect myocardial fibrous structure. AB - Using a three-dimensional propagation model of the human ventricular myocardium, we studied the role of fibrous structure in generating epicardial potential maps. This model represents the myocardium as an anisotropic bidomain with an equal anisotropy ratio, and it incorporates a realistic representation of anatomical features, including epi-endocardial fiber rotation in the compact portion of the wall (compacta) and a distinct fiber arrangement of the trabeculated portion (trabeculata). Activation sequences were elicited at various intramural depths, and maps were calculated throughout a 60 ms sequence. The simulated maps closely resembled those measured by others in the canine heart. During the early stages of activation, a typical map featuring a central minimum flanked by two maxima emerged, with the axis joining these extrema approximately parallel to the fibers near the pacing site, and the axis joining the maxima rotated in the same direction as the fibers for different pacing depths; for endocardial and subendocardial pacing this map changed into one with an oblong positive area. During the later stages of activation, the positive areas of the maps expanded and rotated with the transmural fiber rotation. In concurrence with experiments, we saw a fragmentation and asymmetry of expanding and rotating positive areas. The latter features-apparently caused by the interface between the compacta and trabeculata, variable local thickness of the wall, or local undulations of the vetricular surface-could not be reproduced by more idealized, slab models. PMID- 9846941 TI - Design of a novel apparatus to study nitric oxide (NO) inhibition of platelet adhesion. AB - Nitric oxide (NO) is a simple biological molecule which inhibits adhesion and aggregation of platelets. A novel NO delivery device has been developed to quantitatively study the effects of NO concentration and flux on the adhesion of platelets to a surface. The slit-flow device is lined with a protein-coated membrane through which NO gas permeates into a perfusing platelet suspension. A model predicting spatial NO concentrations and fluxes within the flow slit was validated. At a wall shear rate of 250 s(-1), platelet adhesion was inhibited 87% relative to controls for exposures as low as 0.1 ppm NO. Corresponding model predictions of the aqueous NO concentration and fluxes at the surface were 0.15 nM, and between 0.5 and 1.1 nanomoles cm(-2) s(-1), respectively. Endothelial cells, which release NO to inhibit platelet adhesion in vivo, generate NO at an estimated flux similar to the above values. At a NO exposure of 0.02 ppm, platelet inhibition was only 10%. The delivery device is useful for other studies in which a knowledge of the spatial NO fluxes or concentrations is desired. Knowledge of these fluxes or concentrations is beneficial towards the design of biomaterials incorporating NO to inhibit platelet adhesion. PMID- 9846943 TI - Recording properties and biocompatibility of chronically implanted polymer-based intrafascicular electrodes. AB - We implanted polymer-based longitudinal intrafascicular electrodes (polyLIFEs) in feline dorsal rootlets acutely and for periods of two to six months to evaluate their electrical properties and biocompatibility. A total of 38 implanted electrodes were analyzed. Some 25 of the 38 electrodes were implanted with an insulative flexible polymer cuff, which was required for recording of afferent activity in situ. Electrode impedances remained stable for the duration of the experiments. The distributions of axons were measured at three levels of the implanted rootlets: the implant level, 1-2 mm proximal to the implant with respect to the cell body, and 1-2 mm distal to the implant with respect to the cell body. Similar measurements were made in five samples of fascicles neighboring an implant and six samples of control tissue from animals in which no implants were placed. The polyLIFEs demonstrated a high degree of biocompatibility, as no adverse effects on axon size were observed in either the implanted fascicle or neighboring neural tissue. However, the insulative cuffs were found to be a source of compression, resulting in necrosis of the neural tissue. PMID- 9846942 TI - Enhancement of O2 and CO2 transfer through microporous hollow fibers by pressure cycling. AB - An intravascular gas exchange device for the treatment of respiratory failure consisted of a multitude of blind-ended hollow fibers glued in a pine-needle arrangement to a central gas supply catheter. It has previously been shown that gas desorption rates can be significantly enhanced by cycling gas pressure between a hypobaric level of 130 and an ambient level of 775 Torr. In this study, influences of the cycling frequency (f) and the cycle fraction during which hypobaric pressure is applied (theta) were investigated. Rates of O2 desorption from O2-saturated water and CO2 desorption from CO2-saturated water into a manifold containing 198 fibers, 380 microm in diameter, were measured over a range of f from 0.2 to 1.0 Hz. theta from 0.1 to 0.8, and fiber lengths from 4 to 16 cm. Relative to operation at ambient pressure, pressure cycling increased O2 transfer 3-4 times and CO2 transfer 4-6 times when the water flowed over the fiber manifold at 2.3 l/min. Transfer rates were relatively insensitive tof and theta with 80%-90% of maximum enhancement obtained when theta was as low as 0.2. Transfer rates increased continuously with fiber length, implying that pressure cycling reduced the intra-fiber resistance to gas diffusion. A mathematical diffusion model which utilized only two adjustable parameters, a mass transfer coefficient for O2 and for CO2, simulated the trends exhibited by desorption data. PMID- 9846944 TI - Quantification analysis for saccadic eye movements. AB - Traditional parameters, such as latency and peak velocity, have been used for quantitative analysis of saccade dynamics. However, saccades of equal magnitude which differ substantially in dynamics may still have the same peak velocity. This lack of selectivity might degrade the efficacy of the peak velocity measurement in discriminating patients from normal subjects. In this study, an alternative parameter, the damping ratio of a second-order response, is proposed to concisely and accurately describe saccadic dynamics. A measurement of damping ratio is made by considering the response curve of saccade as the step response of a second-order transfer function. Using the least-mean-square algorithm, the second-order function has been optimized to fit the response of the saccade dynamics with the latency removed. Nineteen normal subjects (ten young and nine older) and 16 patients of Parkinson's disease (eight mildly affected and eight advanced affected) were tested for eye movements in response to pseudorandom saccadic stimuli. While the traditional peak velocity had difficulties in differentiating between the saccadic dynamics of normal and Parkinson subjects, the damping ratio is sensitive enough not only to highlight the difference between a group of patients mildly affected with Parkinson's disease and an age matched normal group (p<0.01; abnormal versus normal), but also to distinguish between groups at different ages (p<0.01; younger versus older). It is proposed that the damping ratio could be a useful parameter in analyzing saccadic dynamics. PMID- 9846945 TI - Continuous wavelet analysis as an aid in the representation and interpretation of electrogastrographic signals. AB - This paper presents an application of the continuous wavelet transform (CWT) in the analysis of electrogastrographic (EGG) signals. Due to the nonstationary nature of EGG signals, the CWT method, which uses multiresolution scaled windows, gives a better time-frequency resolution than the short-time Fourier transform, which uses a fixed window. Spike activity due to gastric contraction was investigated through experiments on dogs. During spike activity we observed an increase in magnitude of the slow wave and the appearance of a low frequency component with half the frequency of the slow wave. Studies of the EGG signals from the small intestine are also presented to investigate the hypothesis that its slow wave might be confounded with spike activity in the stomach due to the similarity of their frequency ranges. PMID- 9846946 TI - Effect of the signal measured from the glottis on determination of the vocal tract shape. AB - All-pole and pole-zero models for the vocal tract are developed. First an impulse train, then the pressure signal measured from the glottis, is used as the input in the models. The models for eight Turkish vowels produced by one male subject are studied to determine the effects of the presumed impulse train and the pressure signal measured from the glottis on the estimation of the vocal tract shape. The motion of the tongue is also examined for a whole word. PMID- 9846948 TI - The versatility of RNA: let's shoot the deputy. PMID- 9846947 TI - Alpha4beta1 and alpha5beta1 control cell migration on fibronectin by differentially regulating cell speed and motile cell phenotype. AB - Computer-aided time lapse fluorescence videomicroscopy was used to study single cell migration behavior of human aortic endothelial cells on fibronectin coated substrates of varying protein surface density. The role of receptors alpha5beta1, alpha(v)beta3, and alpha4beta1 in mediating cell adhesion and migration on fibronectin was characterized using integrin specific monoclonal antibodies. Matrix density had a direct effect on controlling the proportion of migrating cells and the directional persistence of cell movement (p<0.01). While there was relatively little influence of fibronectin surface density on absolute migration speed, the ability of endothelial cells to disperse over a surface, as measured by the dispersion coefficient, was biphasic with respect to the surface density of this matrix protein (p<0.005). Both cell speed and the proportion of migrating cells was controlled by alpha4beta1 (p<0.01). However, alpha5beta1 selectively regulated the transformation of stationary cells to those exhibiting motile behavior (p<0.05). Migratory responses on fibronectin were not influenced by blockade of the alpha(v)beta3 receptor. It is noteworthy that cell surface adhesive receptors which control commitment to a motile phenotype are not necessarily the same as those that control migration speed. PMID- 9846949 TI - Comparative aspects of meiotic cell cycle control in mammals. AB - This review examines the mechanisms of cell cycle control in mammalian germ cells with respect to species-specific variations in reproductive strategies. While sex specific variants are evidenced at the level of checkpoint controls, the timing of meiotic progression, and the modulation of meiosis by hormonal cues, local somatic cell influences impose a hierarchical order to both the initiation and completion of gametogenesis. In the particular case of meiosis in females the rules governing entry into meiotic prophase during gonadal development are generally conserved. In contrast, the regulation of meiotic arrest in developing ovarian follicles, the reinitiation of meiosis at ovulation, and the completion of meiosis upon fertilization involves changes in both the cell cycle machinery and execution of external cues. The overall efficiency of meiotic progression is determined by inputs, mediated by cell contact and/or growth factor, which coordinate oogenesis with folliculogenesis and ensure appropriate and species specific ovulatory outputs (monovular or polyovular). How mechanisms of meiotic cell cycle control can be exploited to improve gamete quality or interfere with fertility is discussed. PMID- 9846950 TI - Germ cell nuclear factor: an orphan receptor with unexpected properties. AB - The orphan receptor germ cell nuclear factor (GCNF) is a member of the superfamily of nuclear receptors. During embryogenesis GCNF expression is restricted to the developing nervous system, whereas in the adult the receptor is also expressed during specific stages in maturing germ cells of the ovary and testis. Therefore GCNF may participate in the regulation of neurogenesis and reproductive functions. Binding of GCNF to the consensus element TCA[AG(G/T)TCA]2 (conRE), to the direct repeat DNA element AGGTCAAGGTCA (DR0), and to extended half-sites such as TCAAGGTCA (XRE) has been demonstrated, but no natural GCNF target gene has been identified. However, due to an overlapping temporal expression pattern and the presence of DR0-type elements in their promoter regions, the protamine 1 and 2 genes have been proposed as potential candidates for a regulation by GCNF. Since GCNF binds as a homodimer to all three elements (conRE, DR0, and XRE) the receptor exhibits an exceptional property within the nuclear receptor superfamily. Homodimeric binding of GCNF to extended half-sites requires the presence of a novel dimerization motif located in the putative helix 3 of the GCNF ligand-binding domain (LBD). Since neither potential ligands nor heterodimerization partners or cofactors for GCNF have been identified, little is known about the mechanisms by which the receptor controls transcriptional processes. Due to the lack of a conserved transcriptional activation function 2 core motif (AF-2 AD core) in the helix 12 region of the GCNF LBD, it has been suggested that GCNF functions as a repressor of transcription. In addition, recent data suggest that the helix 12 region displays functions distinct from those in other nuclear receptors and is involved in the control of DNA binding. Together, these reports indicate that GCNF exhibits novel properties distinct from other members of the nuclear receptor superfamily. PMID- 9846951 TI - Regulating the balance between differentiation and apoptosis: role of CREM in the male germ cells. AB - Various endocrine and neuronal functions are governed by the cAMP-dependent signaling pathway. In eukaryotes, transcriptional regulation upon stimulation of the adenylyl cyclase signaling pathway is mediated by a family of cAMP-responsive nuclear factors. This family consists of a large number of members which may act as activators or repressors. These factors contain the basic domain/leucine zipper motifs and bind as dimers to cAMP-response elements (CRE). The function of CRE-binding proteins (CREBs) is modulated by phosphorylation by several kinases. Direct activation of gene expression by CREBs requires phosphorylation by the cAMP-dependent protein kinase A to the serine-133 residue. The gene CREM encodes various transcription factors which play key physiological and developmental roles within the hypothalamic-pituitary-gonadal axis. We have previously shown that the transcriptional activator CREMtau is highly expressed in postmeiotic cells. Spermiogenesis is a complex process by which postmeiotic male germ cells differentiate into mature spermatozoa. This process involves remarkable structural and biochemical changes which are under the hormonal control of the hypothalamic-pituitary axis. We have addressed the specific role of CREM in spermiogenesis using CREM-mutant mice generated by homologous recombination. Analysis of the seminiferous epithelium from mutant male mice reveals that spermatogenesis stops at the first step of spermiogenesis. Late spermatids are completely absent while there is a significant increase in apoptotic germ cells. A series of postmeiotic germ cell-specific genes are not expressed. Mutant male mice completely lack spermatozoa. This phenotype is reminiscent of cases of human infertility. PMID- 9846952 TI - Meiosis-activating sterols: background, discovery, and possible use. AB - Several years ago we discovered that spent media from cultured human and bull testes contain components that initiate meiosis in germ cells from fetal mouse testes which have been cultured for 6 days in the spent medium. The active substance(s) was termed meiosis-inducing substance. We later found that human follicular fluid harvested after stimulation with gonadotropins has a similar effect. These meiosis-activating substances have now been identified and characterized in extracts from bull testes and human preovulatory follicular fluid as naturally occurring sterols (meiosis-activating sterols, MAS). MAS are intermediates in the cholesterol biosynthetic pathway and are thus present in all cells which produce cholesterol de novo and from lanosterol. However, MAS accumulate only in the gonads. We discuss the possible physiological role of these sterols in initiating meiosis and in oocyte resumption of meiosis, and their potential use in promoting and preventing fertility. PMID- 9846953 TI - Cardiovascular actions of cannabinoids and their generation during shock. AB - Marijuana is a widely abused recreational drug well known for its psychoactive properties. Cannabinoids, the active ingredients of marijuana, elicit their neurobehavioral effects by interacting with the CB1 cannabinoid receptor subtype, expressed primarily in the brain but also present in some peripheral tissues. A second receptor subtype, the CB2 receptor, is expressed on cells of the immune system and is thought to be responsible for the immunosuppressant effects of cannabinoids. Recently, endogenous lipidlike substances have been identified, including arachidonyl ethanolamide (anandamide) and 2-arachidonyl glyceride, that bind to cannabinoid receptors and mimic many of the neurobehavioral effects of plant-derived cannabinoids. Both plant-derived cannabinoids and the endogenous ligands have been shown to elicit hypotension and bradycardia via activation of peripherally located CB1 receptors. Possible underlying mechanisms include presynaptic CB1 receptor mediated inhibition of norepinephrine release from peripheral sympathetic nerve terminals, and/or direct vasodilation via activation of vascular cannabinoid receptors. The latter may also be the target of endocannabinoids of vascular endothelial origin. Recent studies indicate that a peripheral endogenous cannabinoid system in circulating macrophages and platelets is activated in hemorrhagic and septic shock and may contribute to the hypotension associated with these conditions via activation of vascular cannabinoid receptors. The potential role of this mechanism in human shock conditions is under investigation. PMID- 9846954 TI - A novel germline mutation in exon 5 of the multiple endocrine neoplasia type 1 gene. AB - The autosomal dominant multiple endocrine neoplasia type 1 (MEN1) syndrome is characterized by neoplasia of parathyroids, anterior pituitary, and gastrointestinal and pancreatic neuroendocrine tissues. Recently the gene responsible for the MEN1 syndrome has been identified on chromosome region 11q13. Most of the described mutations are nucleotide substitutions and small deletions affecting exons 2 and 3, causing protein truncation. Only one mutation in exon 5 has been found, and this corresponds to a MEN1 sporadic case. Small insertions are also rare. We studied a MENI family composed of five members, two of whom were clinically affected. We found a new germline 1 basepair insertional mutation affecting the exon 5 of the MEN1 gene in the two members affected in this MEN1 family. PMID- 9846955 TI - Detecting every genital papilloma virus infection: what does it mean? PMID- 9846956 TI - Does overexpression of betaAPP in aging muscle have a pathogenic role and a relevance to Alzheimer's disease? Clues from inclusion body myositis, cultured human muscle, and transgenic mice. PMID- 9846957 TI - Transgenic mice over-expressing the C-99 fragment of betaPP with an alpha secretase site mutation develop a myopathy similar to human inclusion body myositis. AB - Inclusion body myositis (IBM) is the most common muscle disease in the elderly. Amyloid-beta protein (A beta) has been shown to accumulate abnormally in the vacuolated fibers and to localize to amyloid-like fibrils in muscles from IBM patients. We studied the skeletal muscles from a line of transgenic mice over expressing the carboxyl-terminal 99 amino acids (C99) of the beta-amyloid precursor protein (betaPP) with a substitution of lysine-612 to valine (K612V), intended to abolish alpha-secretase recognition and to preserve the A beta domain of C99. The majority (87%) of the 24-month-old transgenic mice showed myopathic changes, and approximately one-third of them had degenerating fibers with sarcoplasmic vacuoles and thioflavin-S-positive deposits. Ultrastructurally, the inclusions were aggregates of short thin amyloid-like fibrils, 6 to 8 nm in diameter. These features are similar to those of human IBM. Immunocytochemistry using an antibody against A beta showed membranous staining in most muscle fibers of transgenic mice, as well as granular or vacuolar cytoplasmic staining in the atrophic fibers. Western blots showed a high level of accumulation of carboxyl terminal fragments of betaPP in the muscles of the transgenic mice with the most severe IBM-like lesions. The expression of IBM-like lesions was age dependent. These transgenic mice provide a model for the study of IBM and for the peripheral expression of a key element in the pathogenesis of Alzheimer disease. PMID- 9846958 TI - Amyloid-beta deposition in skeletal muscle of transgenic mice: possible model of inclusion body myopathy. AB - Inclusion body myopathy is a progressive muscle disorder characterized by nuclear and cytoplasmic inclusions and vacuolation of muscle fibers. Affected muscle fibers contain deposits of congophilic amyloid, amyloid-beta immunoreactive filaments, and paired helical filaments, all of which are pathological hallmarks of Alzheimer's disease in brain. Accumulations of amyloid-beta and its precursor are thought to play important roles in the pathogenesis of both inclusion body myopathy and Alzheimer's disease. Overexpression of mutant forms of beta protein precursor in transgenic mice by neuron-specific promoters has been reported to cause amyloid deposits in the brain. Here we report that overexpression in transgenic mice of the signal plus 99-amino acid carboxyl-terminal sequences of beta protein precursor, under the control of a cytomegalovirus enhancer/beta actin promoter, resulted in vacuolation and increasing accumulation of the 4-kd amyloid-beta and the carboxyl-terminus in skeletal muscle fibers during aging. These deposits in transgenic muscle only rarely showed Congo red birefringence. Thus, overexpression of part of beta protein precursor in transgenic mice led to development of some of the characteristic features of inclusion body myopathy. These mice may be a useful model of inclusion body myopathy, which shares a number of pathological markers with Alzheimer's disease. PMID- 9846959 TI - Expression of cellular prion protein in activated hepatic stellate cells. AB - Suppression subtractive hybridization was used to clone genes associated with the activation of hepatic stellate cells and 13 genes were found to be dominantly expressed in activated stellate cells. Among them, one was identical to the 421 837th base pairs of cDNA sequence reported for rat prion-related protein (PrP). In cultured stellate cells, PrP mRNA expression increased in a time-dependent manner in parallel with smooth muscle (SM) alpha-actin mRNA expression. In situ hybridization demonstrated that PrP mRNA was localized in and around the fibrous septa of carbon tetrachloride (CCl4)-treated liver. Cellular PrP (PrPc) was produced by culture-activated stellate cells, and immunohistochemically detected in the fibrous septa of CCl4-damaged liver and sinusoidal linings of common bile duct-ligated liver, consistent with the localization of SM alpha-actin. Immunoelectron microscopy revealed that PrPc resided on the plasma membrane of stellate cells. These results indicate that PrP expression is closely related to stellate cell activation associated with fibrogenic stimuli. PMID- 9846960 TI - Homing receptor alpha4beta7 integrin expression predicts digestive tract involvement in mantle cell lymphoma. AB - Appropriate staging and evaluation of residual disease is critical to improving the treatment of patients with lymphoma. The specific expression of homing receptors may determine the preferential dissemination pattern of tumoral cells. We investigated the expression of the mucosal homing receptor alpha4beta7 on tumoral cells from peripheral lymph node in patients with newly diagnosed mantle cell lymphoma (MCL) to check whether it is associated with gastrointestinal involvement. Expression of the alpha4beta1 integrin and the peripheral lymph node addressin CD62L were also examined. Thirteen MCL patients presenting with peripheral lymphadenopathy were studied. Expression of the mucosal homing receptor integrin alpha4beta7 by peripheral lymph node lymphoma cells was found to be frequent (5/13) and associated with gastrointestinal involvement (5/7). In contrast, lymphoma cells from patients without gastrointestinal involvement did not express alpha4beta7 (6/6) (P = 0.03). These data suggest that alpha4beta7 integrin is expressed by a subset of MCLs and that its expression may predict digestive tract involvement in MCL, furnishing a basis for recognizing two distinct clinical and phenotypic forms, ie, "digestive homing (or digestive primitive)" versus "peripheral" MCL. Further studies on more patients will be needed to understand the impact of biological differences on the prognosis of these two clinical forms. PMID- 9846961 TI - T-cell-rich large-B-cell lymphomas contain non-activated CD8+ cytolytic T cells, show increased tumor cell apoptosis, and have lower Bcl-2 expression than diffuse large-B-cell lymphomas. AB - The factor(s) responsible for the reduced B cell number and increased T cell infiltrate in T-cell-rich large-B-cell lymphomas (TCRBCLs) have not been well characterized. We studied 18 TCRBCLs and 12 diffuse large-B-cell lymphomas (DLBCLs) to compare the 1) predominant T cell subpopulation(s), 2) expression of cytotoxic granule proteins (TIA-1 and granzyme B), 3) level of tumor cell apoptosis (Apoptag system, Oncor, Gaithersburg, MD), and 4) expression of Ki-67 (Mib-1) and apoptosis-related proteins (fas (CD95), bcl-2, and p53). T cells in TCRBCLs and DLBCLs were predominantly CD8+ T cells expressing alphabeta T-cell receptors and TIA-1 (16 of 18 TCRBCLs with >50% TIA-1+ small lymphocytes) but lacking granzyme B (16 of 18 TCRBCLs with <25% granzyme B+ small lymphocytes). Scattered apoptotic tumor cells (confirmed with CD20 co-labeling) were present in 15 of 18 TCRBCLs, with 14 of 15 cases having <10% apoptotic cells. No apoptotic cells were seen in 12 of 12 DLBCLs. In 16 of 16 immunoreactive TCRBCLs, <25% tumor cells were bcl-2+, whereas 6 of 12 DLBCLs had >50% bcl-2+ tumor cells. CD95 (fas) expression was also lower, with 3 of 18 (16.7%) TCRBCLs versus 4 of 12 (33%) DLBCLs having >25% CD95+ tumor cells. TCRBCLs and DLBCLs had similar levels of p53 and Ki-67 (Mib-1) expression. Thus, T cells in TCRBCLs are non-activated cytotoxic T lymphocytes (TIA-1+, granzyme B-). Tumor cell apoptosis (perhaps cytotoxic T cell mediated) may partly account for the decreased number of large (neoplastic) B cells in TCRBCLs, but other factors (ie, decreased bcl-2 expression) may also be needed. PMID- 9846962 TI - Cryptosporidium parvum initiates inflammatory bowel disease in germfree T cell receptor-alpha-deficient mice. AB - Flora-bearing mice with targeted disruption of T cell receptor (TCR)-alpha or beta genes spontaneously develop intestinal inflammation with features similar to ulcerative colitis in humans. TCR-alpha-deficient mice maintained germfree or colonized with a limited number of intestinal bacteria failed to develop inflammatory bowel disease (IBD)-like lesions. Evidently, inflammation in these mice does not develop spontaneously or result from a generalized antigenic stimulation, but rather requires induction by a heretofore unidentified specific stimulus. We describe the development of IBD-like lesions in germfree TCR-alpha deficient mice monoassociated with the protozoan Cryptosporidium parvum. Lesions were seen in distal ileum, cecum, and colon and were most severe in the cecum. A prominent leukocytic infiltrate within the lamina propria was a common characteristic of the lesions observed in the C. parvum-infected germfree TCR alpha-deficient mice. The leukocytic infiltrate was composed of aggregates of B220+ cells, the majority of which expressed surface IgD (ie, conventional B lymphocytes). It has been proposed that antigenic stimulation by a microorganism(s) is needed to initiate intestinal inflammation in TCR-alpha deficient mice. Our results indicate that a single microbial species, C. parvum, is capable of triggering the development of IBD-like lesions in germfree TCR alpha-deficient mice. PMID- 9846963 TI - Eliminating arterial pulsatile strain by external banding induces medial but not neointimal atrophy and apoptosis in the rabbit. AB - We have examined the role of vessel pulsation and wall tension on remodeling and intimal proliferation in the rabbit infrarenal abdominal aorta. A rigid perivascular polyethylene cuff was used to reduce vessel systolic diameter by 25%, producing a region of reduced circumferential strain. At 6 weeks postoperatively, reduced circumferential strain caused medial atrophy, with 45% reduction of medial area and 30% loss of medial smooth muscle cells. Apoptotic cell death was indicated by DNA fragmentation, propidium iodide staining, and cell morphology. Cuffing the aorta after balloon denudation produced medial atrophy but did not inhibit neointimal growth. At 1 week postoperatively, intimal thickness was slightly decreased in regions with reduced strain; however, intimal thickening in regions of reduced strain was not different from control segments at 3 weeks postoperatively (intimal area was 0.37 +/- 0.05 mm2 with reduced strain and 0.50 +/- 0.08 for controls, mean +/- SEM). We conclude that circumferential strain is a major factor controlling medial structure and cell number, whereas growth of the neointima after injury is not significantly affected by either reduced strain or extensive medial cell death. Vessel cuffing represents a new model of blood vessel remodeling in vivo that involves extensive smooth muscle cell apoptosis. PMID- 9846964 TI - Novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses. AB - A novel set of polymerase chain reaction (PCR) primers, designated SPF1 and SPF2 and located in the L1 region, was developed for universal detection of human papillomavirus (HPV). A short PCR fragment (SPF) of only 65 pb was synthesized. SPF amplimers were detected in a microtiter-based hybridization system, using a mixture of oligonucleotide probes. The SPF system allowed detection of at least 43 different HPV genotypes. The clinical performance of the novel SPF system was assessed in three different patient groups. 1) Analysis of 534 cervical scrapes, obtained from treated patients, showed that the detection rate in 447 (83.7%) scrapes with normal cytology was significantly higher using the SPF system as compared with the universal primer set GP5+/6+ (P < 0.001). 2) The SPF assay detected HPV DNA in 299 (98.4%) of 304 scrapes with cytological dyskaryosis. 3) The SPF system detected HPV DNA in 100% of 184 formalin-fixed, paraffin-embedded cervical carcinoma specimens. In conclusion, the novel SPF system permitted universal and highly sensitive detection of HPV DNA in diverse clinical materials and may improve the molecular diagnosis and epidemiology of this important virus. PMID- 9846965 TI - Expression status of p16 protein is associated with human papillomavirus oncogenic potential in cervical and genital lesions. AB - The p16 protein (p16) is a cyclin-dependent kinase (CDK) inhibitor that decelerates the cell cycle by inactivating the CDKs that phosphorylate retinoblastoma (Rb) protein. Recent biological studies have revealed that p16 expression is markedly influenced by the status of Rb expression, and p16 overexpression has been demonstrated in cervical cancers because of functional inactivation of Rb by human papillomavirus (HPV) E7 protein. To clarify the relationship between p16 overexpression and HPV infection in cervical carcinogenesis, immunohistochemical analysis of p16 and detection of HPV by in situ hybridization and polymerase chain reaction were performed on 139 formalin fixed and paraffin-embedded samples of cervical and genital condylomatous and neoplastic lesions. Marked overexpression of p16 protein, ie, diffuse and strong immunostaining, was observed in all cervical cancers and preneoplastic lesions with infection by high- and intermediate-risk HPVs, ie, subtypes 16, 18, 31, 33, 52, and 58. Condylomata acuminata and low-grade squamous intraepithelial lesions with infection by low-risk HPV such as HPV-6/11 showed focal and weak immunohistochemical staining for p16. Our results clearly showed that the mode of p16 expression in lesions with high- and intermediate-risk HPVs differed from its expression in lesions with low-risk HPVs and thus might be attributable to differences in functional inactivation of Rb protein by different HPVs. PMID- 9846967 TI - Loss of the tight junction MAGUK ZO-1 in breast cancer: relationship to glandular differentiation and loss of heterozygosity. AB - Membrane-associated guanylate kinase homologs (MAGUKs) may play a role in cellular functions preventing tumorigenesis as indicated by the neoplastic phenotype caused by genetic loss of the MAGUK Dlg in Drosophila. To test this possibility, we examined the expression and subcellular localization of the tight junction MAGUK ZO-1, as well as the cell adhesion molecule E-cadherin, in paraffin-embedded breast cancer samples, using immunohistochemistry and confocal microscopy. As expected, normal tissue showed intense staining for ZO-1 at the position of the epithelial tight junctions, but this staining was reduced or lost in 69% of breast cancers analyzed (n = 48). In infiltrating ductal carcinomas (n = 38) there was a reduction in staining in 42% of well differentiated, in 83% of moderately differentiated and 93% of poorly differentiated tumors. ZO-1 staining was positively correlated with tumor differentiation (P = .011) and more specifically with the glandular differentiation of tumors (P = .0019). Reduction in ZO-1 staining was strongly correlated with reduced E-cadherin staining (P = 4.9 x 10(-5)). The results suggest that down-regulation of ZO-1 expression and its failure to accumulate at cell junctions may be causally related to cancer progression. To detect loss of heterozygosity, the ZO-1 gene tjp-1 was mapped relative to other markers in 15q13 and polymorphic markers flanking tjp-1 were identified. The marker D15S1019 showed loss of heterozygosity in 23% of informative tumors (n = 13). Loss of a tjp-1-linked marker suggests that genetic loss may, in some cases, be responsible for the reduction in ZO-1 expression in breast cancer. PMID- 9846966 TI - Alterations of the p16-pRb pathway and the chromosome locus 9p21-22 in non-small cell lung carcinomas: relationship with p53 and MDM2 protein expression. AB - The p16-pRb and p53-MDM2 pathways represent vital cell cycle checkpoints. Recent studies provide evidence that these pathways are directly linked via MDM2-pRb interaction and p53 suppression of the RB1 gene. In the present study we investigated the alterations of this G1 phase protein network using immunohistochemical and molecular methods in a series of 68 non-small-cell lung carcinomas (NSCLCs) and correlated the findings with clinicopathological features and prognosis of the patients. Aberrant expression (Ab) of p16 and pRb was observed in 33 (49%) and 27 (40%) of the carcinomas, respectively. Analysis of the region that encodes for p16 by deletion mapping, a polymerase chain reaction (PCR)-based methylation assay and PCR single-strand conformation polymorphism (SSCP) analysis revealed that deletions and transcriptional silencing by methylation might represent the main mechanisms of CDKN2/p16ink4a inactivation in NSCLCs. The results of deletion mapping also suggest that other tumor suppressor genes may reside at the 9p21-22 region, which encodes for CDKN2/MTS1/p16ink4a, p14ARF, and MTS2/p15ink4b. In addition, microsatellite instability was observed with a frequency of 16% in the 9p21-22 chromosome area. Overexpression (P) of p53 and MDM2 proteins was found in 39 (58%) and 47 (70%) of the cases, respectively. A highly significant association was observed between p53 overexpression and p53 mutations (P = 0.006). Statistical analysis of the expression patterns of the biologically relevant molecules (p16/pRb, p53/MDM2, MDM2/pRb, and p53/pRb) showed coincident overexpression of p53 and MDM2 (P = 0.04) and that abnormal pRb was correlated with elevated levels of MDM2 (P = 0.013) and p53 (P = 0.01), respectively. We suggest that deregulated expression of these molecules may act synergistically. An important finding of the study was that multiple impairments (three and four molecules affected) of the p16/pRb/p53/MDM2 network occurred in a large proportion (43%) of the carcinomas. This finding in addition to the absence of correlation with clinical stage of the tumors suggests that multiple hits of this network may be a relatively early event in the development of a subset of NSCLCs. The relationship between the factors examined in the present study, clinicopathological features, and survival of the patients did not reveal any significant correlations with the exception of smoking, which was associated with microsatellite alterations (loss of heterozygosity and microsatellite instability) at the 9p21-22 locus (P = 0.04) and the immunophenotypes p53(P)/MDM2(P) (P = 0.04) and p16(Ab)/pRb(Ab)/p53(P)/MDM2(P) (P = 0.03), respectively. We suggest that in a subset of NSCLCs, simultaneous deregulation of the members of this network may represent one way of initiating the oncogenic procedure whereas in other NSCLC subgroups alternative pathways may play this role. PMID- 9846968 TI - Arsenic enhancement of skin neoplasia by chronic stimulation of growth factors. AB - Although numerous epidemiological studies have shown that inorganic arsenicals cause skin cancers and hyperkeratoses in humans, there are currently no established mechanisms for their action or animal models. Previous studies in our laboratory using primary human keratinocyte cultures demonstrated that micromolar concentrations of inorganic arsenite increased cell proliferation via the production of keratinocyte-derived growth factors. As recent reports demonstrate that overexpression of keratinocyte-derived growth factors, such as transforming growth factor (TGF)-alpha, promote the formation of skin tumors, we hypothesized that similar events may be responsible for those associated with arsenic skin diseases. Thus, the influence of arsenic in humans with arsenic skin disease and on mouse skin tumor development in transgenic mice was studied. After low-dose application of tetradecanoyl phorbol acetate (TPA), a marked increase in the number of skin papillomas occurred in Tg.AC mice, which carry the v-Ha-ras oncogene, that received arsenic in the drinking water as compared with control drinking water, whereas no papillomas developed in arsenic-treated transgenic mice that did not receive TPA or arsenic/TPA-treated wild-type FVB/N mice. Consistent with earlier in vitro findings, increases in granulocyte/macrophage colony-stimulating factor (GM-CSF) and TGF-alpha mRNA transcripts were found in the epidermis at clinically normal sites within 10 weeks after arsenic treatment. Immunohistochemical staining localized TGF-alpha overexpression to the hair follicles. Injection of neutralizing antibodies to GM-CSF after TPA application reduced the number of papillomas in Tg.AC mice. Analysis of gene expression in samples of skin lesions obtained from humans chronically exposed to arsenic via their drinking water also showed similar alterations in growth factor expression. Although confirmation will be required in nontransgenic mice, these results suggest that arsenic enhances development of skin neoplasias via the chronic stimulation of keratinocyte-derived growth factors and may be a rare example of a chemical carcinogen that acts as a co-promoter. PMID- 9846969 TI - Pituitary adenylate-cyclase-activating polypeptide (PACAP) binding sites and PACAP/vasoactive intestinal polypeptide receptor expression in human pituitary adenomas. AB - Pituitary adenylate-cyclase-activating polypeptide (PACAP) stimulates release of several anterior pituitary hormones by interacting with PACAP receptors on pituitary cells. To learn more about the distribution and possible regulatory roles of PACAP and its receptors in human pituitary adenomas, we investigated the expression of vasoactive intestinal polypeptide (VIP) and PACAP binding sites using receptor autoradiography, PACAP and PACAP/VIP receptor (PVR) mRNAs by reverse transcription polymerase chain reaction (RT-PCR), conventional in situ hybridization, and catalyzed reporter deposition in situ hybridization (CARD-ISH) analyses. PACAP mRNA was expressed in normal human hypothalamus, which was used as a positive control, but not in pituitary adenomas. Receptor autoradiography showed PACAP types I and II binding sites in all groups of pituitary adenomas, except prolactinomas. The highest levels were present in gonadotroph and null cell adenomas. PVR-2 mRNA was expressed in normal pituitaries and in all groups of pituitary adenomas by RT-PCR, whereas PVR-1 and -3 mRNAs were expressed in all groups of pituitary adenomas, except for most prolactinomas. Conventional in situ hybridization studies with digoxigenin-labeled probes demonstrated weak staining for PVR-1, -2, and -3 mRNAs in most tissues. The CARD-ISH technique, which increased the sensitivity of the in situ hybridization method, also revealed PVR 2 mRNA expression in all adenomas, whereas PVR-1 and -3 mRNAs were detected in nearly all adenomas except for prolactinomas. The presence of PACAP mRNA in the hypothalamus, but not in normal anterior pituitary or in pituitary adenomas, and the differential expression of PVRs in adenomas indicate a selective regulatory endocrine and paracrine role of PACAP in normal and neoplastic anterior pituitary cells. PMID- 9846970 TI - Selection of potentially metastatic subpopulations expressing c-erbB-2 from breast cancer tissue by use of an extravasation model. AB - Overexpression of the crbB-2 gene-encoded p185(c-erbB-2) is correlated with early onset of metastasis in breast cancer patients. Furthermore, the detection of blood-borne epithelium-derived clustered cells expressing p185(c-erbB-2) was related to advanced stages in breast cancer. To further elucidate the receptor's function in the metastatic process of human breast cancers, we analyzed disaggregated cells and cell clusters from freshly dissected breast cancer tissues. We studied whether their capability of extravasation is correlated with their expression of c-erbB-2. A model for the venular wall was constructed by growing human umbilical vein endothelial cells (HUVECs) on porous membranes coated with basement membrane extracellular matrix. In four control breast cancer cell lines (SK-BR-3, MCF-7, MDA-MB-468, and MDA-MB-468, the latter transfected with a full-length c-erbB-2 cDNA vector) producing different levels of the c-erbB 2 receptor, the expression level correlated positively with the invasiveness of the cells. The invasive, predominantly clustered cells from 14 of 23 tumors were positively stained for p185(c-erbB-2) by immunocytochemistry. Furthermore, we show that the invasive cell populations express the metastasis-associated proteins matrix metalloproteinase MMP-2, CD44, and integrins alpha(v)beta3 and alpha6. In this first study on the behavior of cells and cell clusters from disaggregated operated cancers in an extravasation model, we could demonstrate the presence of c-erbB-2-expressing cell subpopulations within the individual breast cancers that are presumably of high metastatic potential. PMID- 9846971 TI - Detection of SYT-SSX fusion transcripts in synovial sarcoma by reverse transcription-polymerase chain reaction using archival paraffin-embedded tissues. AB - The reciprocal translocation t(X;18)(p11;q11) is known to be highly characteristic of synovial sarcoma, and its consequence, an SYT-SSX fusion gene, is expected to be a diagnostic molecular marker. In this study, we conducted a reverse transcription-polymerase chain reaction-based assay to detect the SYT-SSX fusion gene transcripts using archival formalin-fixed, paraffin-embedded tumor specimens from a series of 32 synovial sarcoma cases including 6 tumors found in unusual anatomical sites. The SYT-SSX fusion transcripts could be detected in 30 of 32 paraffin-embedded specimens (94%). A subsequent sequence analysis using the polymerase chain reaction products confirmed that the detected messages were derived from either the SYT-SSX1 (22 cases) or SYT-SSX2 (8 cases) fusion gene. Of 23 SYT-SSX-positive monophasic tumors, 16 tumors had an SYT-SSX1 fusion transcript. Fusion transcripts were detectable in all the 7 biphasic tumors analysed, one of which had an SYT-SSX2 fusion transcript. All of the six tumors at unusual locations (lung; 3, metastasis to the abdominal cavity from a tumor of retroperitoneal origin; 1, sacral region; 1, iliopsoas muscle; 1) contained detectable messages. Our results indicate that this molecular assay can be applied to archival formalin-fixed, paraffin-embedded tumor tissues as a feasible and reliable molecular technique for the diagnosis of synovial sarcoma. PMID- 9846972 TI - T cell apoptosis in human heart allografts: association with lack of co stimulation? AB - It is unclear whether the intracardial immune reactivity after heart transplantation influences the peripheral immunological status (activation or nonresponsiveness) of the patient. Co-stimulation and activation-induced cell death (AICD) or apoptosis play an important role in determining the balance between lymphocyte reactivity and nonreactivity. Therefore, we studied the expression of co-stimulatory molecules and the process of apoptosis in biopsies of human heart allografts, using immunohistochemistry. Although a normal expression of co-stimulatory molecules on antigen-presenting cells was observed, the expression of their counter-structures on T cells was absent. This may be due to chronic T cell activation, which can lead to the induction of apoptosis via the Fas/Fas ligand pathway. In the infiltrates, a considerable percentage of the lymphocytes, but not the macrophages, were apoptotic. Apoptosis was confirmed by DNA fragmentation analysis. Increased numbers of Bax-expressing versus decreased numbers of Bcl2-expressing lymphocytes in comparison with normal lymphoid tissue confirmed a imbalance in favor of apoptosis. Apoptosis was biased towards CD4+ T cells (65.7% versus 26.6% in CD8+ T cells). Fas was expressed on most of the infiltrating cells. Fas ligand expression was also observed, not only on most of the T cells but also on all macrophages. Because macrophages were often detected in close contact with T cells, they may play a role in T cell regulation via the Fas/Fas ligand pathway. This study indicates that, during rejection, not only is tissue damage induced by infiltrating T cells, but also the infiltrating lymphocytes themselves are actively down-regulated (eg, AICD) by one another and by macrophages in the infiltrate. This regulatory process may affect the immunological status of the patient after heart transplantation. PMID- 9846973 TI - ACE inhibitor quinapril reduces the arterial expression of NF-kappaB-dependent proinflammatory factors but not of collagen I in a rabbit model of atherosclerosis. AB - Increasing evidence supports an association between inflammation and plaque rupture. Macrophages and vascular smooth muscle cells are a source of cytokines and growth factors, which contribute to ongoing inflammation during atherogenesis. In a rabbit model of atherosclerosis, we evaluated the effect of the ACE inhibitor quinapril on different parameters implicated in the pathogenesis of the plaque, such as the presence of chemokines (interleukin-8, monocyte chemoattractant protein-1), collagen I, and vascular smooth muscle cell proliferation (PDGF-B). Since nuclear factor kappaB (NF-kappaB) has been implicated in the control of chemokine transcription and cell proliferation, we also investigated its activation and localization in the lesion. Quinapril administration for 28 days caused a down-regulation in arterial expression of interleukin-8 and monocyte chemoattractant protein-1 (mRNA and protein). However, collagen I expression (mRNA and protein) was not modified. PDGF-B expression was reduced in both the intima and the media. Active NF-kappaB, found in both macrophages and vascular smooth muscle cells, was also reduced by quinapril. Nevertheless, no significant changes were noted in the mild neointima formation, although a certain trend toward normalization was found in the quinapril-treated group. In conclusion, our results show that quinapril treatment attenuates several parameters associated with inflammation within the atherosclerotic lesions that are controlled by NF-kappaB, although it has no effect on collagen I expression. Both effects could contribute to the stabilization of the atherosclerotic plaque. PMID- 9846974 TI - TNF-alpha receptor knockout mice are protected from the fibroproliferative effects of inhaled asbestos fibers. AB - We have demonstrated that C57BL/6-129 hybrid mice with genes for both the 55kd and 75kd receptors for TNF-alpha knocked out (TNF-alphaRKO) fail to develop fibroproliferative lesions after asbestos exposure. There is good evidence that TNF-alpha plays a major role in mediating interstitial pulmonary fibrosis. Our findings support this view and we present here new data obtained by in situ hybridization showing that expression of the genes coding for transforming growth factor alpha (TGF-alpha) and platelet-derived growth factor A-chain (PDGF-A) is reduced in the TNF-alphaRKO mice compared with control animals. In accordance with this observation, data on bromodeoxyuridine (BrdU) incorporation in the lungs of the TNF-alphaRKO mice show no increases over unexposed control animals. In contrast, wild-type control mice exposed to asbestos exhibit 15- to 20-fold increases in BrdU uptake and consequently develop fibrogenic lesions. Even though the levels of TNF-alpha gene expression and protein production were increased in the asbestos-exposed TNF-alphaRKO mice, the lack of receptor signaling protected the mice from developing fibroproliferative lesions. We agree with the view that TNF-alpha is essential for the development of interstitial pulmonary fibrosis and postulate that TNF-alpha mediates its effects through activation of other growth factors such as PDGF and TGF-alpha that control cell growth and matrix production. PMID- 9846975 TI - Chemokines IL-8, GROalpha, MCP-1, IP-10, and Mig are sequentially and differentially expressed during phase-specific infiltration of leukocyte subsets in human wound healing. AB - Healing of cutaneous wounds requires a complex integrated network of repair mechanisms, including the action of newly recruited leukocytes. Using a skin repair model in adult humans, we investigated the role chemokines play in sequential infiltration of leukocyte subsets during wound healing. At day 1 after injury, the C-X-C chemokines IL-8 and growth-related oncogene alpha are maximally expressed in the superficial wound bed and are spatially and temporally associated with neutrophil infiltration. IL-8 and growth-related oncogene alpha profiles also correlate with keratinocyte migration and subsequently subside after wound closure at day 4. Macrophage infiltration reaches the highest levels at day 2 and is paralleled by monocyte chemoattractant protein-1 mRNA expression in both the basal layer of the proliferative epidermis at the wound margins and mononuclear cells in the wound area. Other monocyte-attracting chemokines such as monocyte chemoattractant protein-3, macrophage inflammatory protein-1alpha and 1beta, RANTES, and 1309 are undetectable. At day 4, perivascular focal lymphocyte accumulation correlates with strong focal expression of the C-X-C chemokines Mig and IP-10. Our results suggest that a dynamic set of chemokines contributes to the spatially and temporally different infiltration of leukocyte subsets and thus integrates the inflammatory and reparative processes during wound repair. PMID- 9846976 TI - Collagen deposition in a non-fibrotic lung granuloma model after nitric oxide inhibition. AB - Recent studies support the concept that pulmonary granulomatous inflammation directed by interferon (IFN)-gamma, interleukin (IL)-12, and nitric oxide usually resolves in the absence of fibrosis. To determine whether nitric oxide participates in modulating the fibrotic response during the development of pulmonary granulomas in response to purified protein derivative (PPD), mice presensitized to PPD received daily intraperitoneal injections of N(G)-nitro-D arginine-methyl ester (D-NAME), N(G)-nitro-L-arginine-methyl ester (L-NAME), or aminoguanidine after delivery of PPD-coated beads to the lungs. Eight days later, morphometric analysis of lung granulomas revealed that L-NAME-treated mice when challenged with PPD in vitro for 36 hours had the largest pulmonary granulomas and the greatest collagen deposition among the treated groups. In addition, equivalent numbers of dispersed lung cells from L-NAME- and aminoguanidine treated mice produced significantly higher levels of IL-4, monocyte chemoattractant protein (MCP)-1, and macrophage inflammatory protein (MIP)-1alpha and significantly lower levels of eotaxin compared with D-NAME-treated mice. Cultures of dispersed lung cells from L-NAME-treated mice also produced significantly more IL-10 and less IL-12 compared with similar numbers of dispersed lung cells from D-NAME-treated mice. Cultures of isolated lung fibroblasts from L-NAME-treated mice expressed higher levels of C-C chemokine receptor 2 (CCR2) and CCR3 mRNA and contained less MCP-1 and eotaxin protein than a similar number of fibroblasts from D-NAME-treated mice. Thus, nitric oxide appears to regulate the deposition of extracellular matrix in lung granulomas through the modulation of the cytokine and chemokine profile of these lesions. Alterations in the cytokine, chemokine, and procollagen profile of this lesion may be a direct effect of nitric oxide on the pulmonary fibroblast and provide an important signal for regulating fibroblast activity during the evolution of chronic lung disease. PMID- 9846977 TI - Acute effects of inhaled urban particles and ozone: lung morphology, macrophage activity, and plasma endothelin-1. AB - We studied acute responses of rat lungs to inhalation of urban particulate matter and ozone. Exposure to particles (40 mg/m3 for 4 hours; mass median aerodynamic diameter, 4 to 5 microm; Ottawa urban dust, EHC-93), followed by 20 hours in clean air, did not result in acute lung injury. Nevertheless, inhalation of particles resulted in decreased production of nitric oxide (nitrite) and elevated secretion of macrophage inflammatory protein-2 from lung lavage cells. Inhalation of ozone (0.8 parts per million for 4 hours) resulted in increased neutrophils and protein in lung lavage fluid. Ozone alone also decreased phagocytosis and nitric oxide production and stimulated endothelin-1 secretion by lung lavage cells but did not modify secretion of macrophage inflammatory protein-2. Co exposure to particles potentiated the ozone-induced septal cellularity in the central acinus but without measurable exacerbation of the ozone-related alveolar neutrophilia and permeability to protein detected by lung lavage. The enhanced septal thickening was associated with elevated production of both macrophage inflammatory protein-2 and endothelin-1 by lung lavage cells. Interestingly, inhalation of urban particulate matter increased the plasma levels of endothelin 1, but this response was not influenced by the synergistic effects of ozone and particles on centriacinar septal tissue changes. This suggests an impact of the distally distributed particulate dose on capillary endothelial production or filtration of the vasoconstrictor. Overall, equivalent patterns of effects were observed after a single exposure or three consecutive daily exposures to the pollutants. The experimental data are consistent with epidemiological evidence for acute pulmonary effects of ozone and respirable particulate matter and suggest a possible mechanism whereby cardiovascular effects may be induced by particle exposure. In a broad sense, acute biological effects of respirable particulate matter from ambient air appear related to paracrine/endocrine disruption mechanisms. PMID- 9846978 TI - Bradykinin stimulates type II alveolar cells to release neutrophil and monocyte chemotactic activity and inflammatory cytokines. AB - In the present study, we evaluated the potential of bradykinin (BK) to induce the release of neutrophil and monocyte chemotactic activity (NCA and MCA) and cytokines from an alveolar type II epithelial cell line, A549 cells. BK stimulated A549 cells to release NCA and MCA in a dose- and time-dependent manner (P < 0.001). Checkerboard analysis revealed that both NCA and MCA involved chemotactic and chemokinetic activity. Molecular sieve column chromatography showed three molecular weight masses (near 19 kd, 8 kd, and 400 d) for NCA and several molecular weight peaks (near 66 kd, 25 kd, 19 kd, 16 kd, and 400 d) for MCA. The release of NCA and MCA was inhibited by cycloheximide and lipoxygenase inhibitors (P < 0.01). The NCA and MCA were inhibited by leukotriene B4 (LTB4) receptor antagonist (P < 0.01), and the concentration of LTB4 was high enough for NCA and MCA. Antibodies to interleukin (IL)-8 and granulocyte colony-stimulating factor (G-CSF) attenuated NCA (P < 0.01), and antibodies to monocyte chemotactic protein-1 (MCP-1), G-CSF, and transforming growth factor (TGF)-beta attenuated MCA (P < 0.01). The levels of IL-8, G-CSF, MCP-1, and TGF-beta increased time dependently (P < 0.01). BK also stimulated the release of ILeukin-6 from A549 cells (P < 0.001). The receptors responsible for the release of NCA, MCA, and individual chemokines involved both BKB1 and BKB2 receptors. These data suggest that BK may stimulate alveolar type II pneumocytes to release inflammatory cytokines, which then may modulate the lung inflammation. PMID- 9846979 TI - Altered balance between matrix metalloproteinases and their inhibitors in experimental biliary fibrosis. AB - A rat model of common bile duct ligation (BDL)-induced hepatic fibrosis was used to assess the expression and activities of collagen-degrading proteinases and their inhibitors during the progression of fibrosis. Expression of four members of the matrix metalloproteinase (MMP) family (MMP-2/gelatinase A, MMP-3, MMP 9/gelatinase B, and MMP-13) and three tissue inhibitors of metalloproteinases-1, 2, and -3 (TIMP-1, TIMP-2, and TIMP-3) were evaluated by Northern blot analysis of RNA from liver tissue isolated at 0, 2, 5, 10, 20, and 30 days after either a BDL or sham operation. In addition, we analyzed free gelatinase and TIMP activities by zymography and reverse zymography, respectively. We found that the proteolytic activities of MMP-2 and MMP-9 increased by 2 days after ligation, reached maximal levels at day 10, and remained high through the study period, whereas the gelatinolytic activities in plasma were unchanged. The increase in gelatinase activities was accompanied by an increase in the TIMP mRNA transcripts. TIMP-1 transcripts appeared at day 2, increased until day 10, and remained elevated throughout the study period. TIMP-2 and TIMP-3 transcripts become detectable on day 10 and remained stable afterwards. No corresponding increase in TIMP protein activity was detected by reverse zymography. This appears to result from the formation of TIMP/MMP complexes. These findings indicate a likely surplus in the BDL model of fibrosis of free gelatinases as compared with the TIMPs. Thus, excessive TIMP production is not a sufficient explanation for the observed extracellular matrix accumulation, but complex changes in the local MMP/TIMP balance may underlie the pathomechanisms of fibrosis. PMID- 9846980 TI - Smoke extract stimulates lung epithelial cells to release neutrophil and monocyte chemotactic activity. AB - Inflammatory cells accumulate within the lungs of cigarette smokers. Current concepts suggest that these cells can induce protease-antiprotease and/or oxidant antioxidant imbalance(s), which may damage the normal lung alveolar and interstitial structures. Because type II pneumocytes line the alveolar space, and because the inflammatory cells migrate and reside at the alveolus, we postulated that the type II pneumocytes might release chemotactic activity for neutrophils and monocytes in response to smoke extract. To test this hypothesis, A549 cells were cultured and the supernatant fluids were evaluated for the neutrophil and monocyte chemotactic activity (NCA and MCA) by a blind-well chamber technique. A549 cells released NCA and MCA in response to smoke extract in a dose- and time dependent manner (P < 0.05). Checkerboard analysis showed that the activity was chemotactic. Partial characterization of NCA and MCA revealed that the activity was partly heat labile, trypsin sensitive, and ethyl acetate extractable. Lipoxygenase inhibitors and cycloheximide inhibited the release of NCA and MCA. Molecular sieve column chromatography showed multiple peaks for both NCA and MCA. NCA was inhibited by anti-human-interleukin (IL)-8 antibody, granulocyte colony stimulating factor (G-CSF) antibody, or leukotriene (LT)B4 receptor antagonist. Monocyte chemoattractant protein (MCP)-1 antibody or LTB4 receptor antagonist inhibited MCA. Immunoreactive IL-8, G-CSF, MCP-1, and LTB4 significantly increased in the supernatant fluids in response to smoke extract. These data suggest that the type II pneumocytes may release NCA and MCA and modulate the inflammatory cell recruitment into the lung. PMID- 9846981 TI - Spontaneous neoplastic transformation of WB-F344 rat liver epithelial cells. AB - Several studies have shown that cultured rat liver epithelial cells transform spontaneously after chronic maintenance in a confluent state in vitro. In the present study, multiple independent lineages of low-passage WB-F344 rat liver epithelial stem-like cells were initiated and subjected in parallel to selection for spontaneous transformation to determine whether spontaneous acquisition of tumorigenicity was the result of events (genetic or epigenetic) that occurred independently and stochastically, or reflected the expression of a pre-existing alteration within the parental WB-F344 cell line. Temporal analysis of the spontaneous acquisition of tumorigenicity by WB-F344 cells demonstrated lineage specific differences in the time of first expression of the tumorigenic phenotype, frequencies and latencies of tumor formation, and tumor differentiations. Although spontaneously transformed WB-F344 cells produced diverse tumor types (including hepatocellular carcinomas, cholangiocarcinomas, hepatoblastomas, and osteogenic sarcomas), individual lineages yielded tumors with consistent and specific patterns of differentiation. These results provide substantial evidence that the stochastic accumulation of independent transforming events during the selection regimen in vitro were responsible for spontaneous neoplastic transformation of WB-F344 cells. Furthermore, cell lineage commitment to a specific differentiation program was stable with time in culture and with site of transplantation. This is the first report of a cohort of related, but independent, rat liver epithelial cell lines that collectively produce a spectrum of tumor types but individually reproduce a specific tumor type. These cell lines will provide valuable reagents for investigation of the molecular mechanisms involved in the differentiation of hepatic stem-like cells and for examination of potential causal relationships in spontaneously transformed rat liver epithelial cell lines between molecular/cellular alterations and the ability to produce tumors in syngeneic animals. PMID- 9846982 TI - Lipoprotein receptors in acute myelogenous leukemia: failure to detect increased low-density lipoprotein (LDL) receptor numbers in cell membranes despite increased cellular LDL degradation. AB - The high-affinity degradation of low-density lipoprotein (LDL) is enhanced 3- to 100-fold in leukemic blood cells from patients with acute myelogenous leukemia (AML), suggesting an increased cellular LDL receptor expression. There are, however, inconsistencies regarding the published properties of LDL receptor regulation in AML cells, and previous data on this are indirect. In the present study the aim was to determine whether the LDL receptor number is increased in AML cells. The LDL receptor number was assayed by ligand blot with rabbit 125I labeled beta-very-low-density lipoprotein (beta-VLDL) of transferred, SDS polyacrylamide-gel-electrophoresis-separated AML cell membranes. Samples from 10 patients, six with AML, one with chronic myelogenous leukemia in blast crisis, and three with acute lymphoblastic leukemia, were investigated. The LDL receptor expression was strongly suppressed in all samples to levels lower than that of normal mononuclear cells. This was despite the fact that cells from one patient with AML of M4 subtype had a 50- to 100-fold higher 125I labeled LDL degradation compared with normal cells. Immunoblots with antibodies against gp330/megalin and the LDL-receptor-related protein (LRP) and ligand blot using 125I-labeled 39-kd receptor-associated protein (RAP) could not detect gp330/megalin or VLDL receptors. The LRP was abundant in AML samples of M4 and M5b subtype, as determined from both RAP ligand blot and immunoblot using an LRP-specific antibody. It is concluded that LDL receptors are suppressed in AML cells. It is possible that the high degradation of 125I-labeled LDL present in type M4 and M5 AML cells may involve another lipoprotein receptor. PMID- 9846984 TI - Fas ligand is present in tumors of the Ewing's sarcoma family and is cleaved into a soluble form by a metalloproteinase. AB - Fas ligand (FasL) exists in transmembrane and soluble forms and induces apoptosis on cross-linking with the Fas receptor. We evaluated the biological significance of FasL and Fas in 61 tumor tissues and 9 cell lines of the Ewing's sarcoma family of tumors (ESFT). FasL was present in 62.5% and Fas in 79.4% of primary ESFT. Metastatic tumors had higher expression of FasL (95%), suggesting association with a metastatic phenotype. FasL was detected in the cytoplasm and membrane of ESFT cells by immunofluorescence. Western blotting revealed transmembrane and soluble FasL in cytosolic extracts and soluble FasL in conditioned media. Both transmembrane and soluble FasL induced apoptosis of Fas sensitive Jurkat cells in co-culture experiments with ESFT cells or their media. Treatment with phenanthroline and the synthetic metalloproteinase inhibitor BB 3103 reduced the levels of soluble FasL in the media, suggesting that in ESFT, FasL is processed by a metalloproteinase and released in the extracellular milieu. The released soluble FasL may serve to attack cells of the immune system and/or interfere with the binding of transmembrane FasL with Fas, and results in down-regulation of transmembrane FasL. Synthetic metalloproteinase inhibitors may modify the ratio of transmembrane to soluble FasL. PMID- 9846983 TI - Engagement of CD99 induces apoptosis through a calcineurin-independent pathway in Ewing's sarcoma cells. AB - Programmed cell death (PCD) is a prominent feature of the development of the immune and nervous systems. In both systems, widespread PCD occurs in primitive progenitor cells during development. In this study, we demonstrated that Ewing's sarcoma (ES) cells, undifferentiated neural precursors, underwent apoptosis upon engagement of CD99 with anti-CD99 monoclonal antibody. Apoptosis via CD99 occurred only in the undifferentiated state of ES cells, but not in differentiated ES cells. CD99-induced apoptosis in ES cells appeared to require de novo synthesis of RNA and protein as well as caspase activation. Cyclosporin A, known to be a potent inhibitor of both calcineurin activation and mitochondrial permeability transition pore opening, inhibited CD99-mediated apoptosis, whereas FK-506, a specific calcineurin inhibitor, did not, indicating the induction of CD99-mediated apoptosis through a calcineurin-independent pathway. Furthermore, the dying cells displayed the reduction of mitochondrial transmembrane potential (delta psi m). These results suggest that CD99 engagement induce CsA-inhibitable mitochondrial permeability transition pore opening, followed by a reduction of delta psi m and caspase activation, thereby leading to apoptosis. Based on these results, we suggest the possible involvement of CD99 in the apoptotic processes that occur during nervous system development and also its application in immunotherapeutic trials for ES cases. PMID- 9846985 TI - Uroplakin gene expression by normal and neoplastic human urothelium. AB - cDNA sequences for human uroplakins UPIa, UPIb, UPII, and UPIII were cloned and used to investigate uroplakin transcription by normal and neoplastic urothelial cells. Normal urothelium expressed mRNA for all four uroplakins, although UPIII could be detected only by ribonuclease protection assay. By in situ hybridization, UPIa and UPII were confined to superficial cells and UPIb was also expressed by intermediate cells. Cultured normal human urothelial cells showed a proliferative basal/intermediate cell phenotype and constitutive expression of UPIb only. Uroplakin expression by transitional cell carcinoma cell lines was related to their differentiated phenotype in vitro. RT4 cells expressed all uroplakins, VM-CUB-3 expressed three uroplakins, RT112 and HT1376 cells expressed only UPIb in high abundance, and COLO232, KK47, and EJ cells had no detectable expression. These results correlated with patterns of uroplakin expression in tumors. UPIa and UPII were detected superficially only in well differentiated transitional cell carcinoma papillae. UPIb was positive in seven of nine and overexpressed in five of nine noninvasive transitional cell carcinomas and was also present in four of eight invasive transitional cell carcinomas. Lymph node metastases retained the same pattern of UPIb expression as the primary tumor. Unlike the three differentiation-regulated uroplakins, UPIb may have an alternative role in urothelial cell/tissue processes. PMID- 9846986 TI - Differential expression of cyclin D1 in mantle cell lymphoma and other non Hodgkin's lymphomas. AB - Mantle-cell lymphomas are associated with a characteristic chromosomal translocation, t(11;14)(q13;q32). This translocation involves rearrangement of the bcl-1 proto-oncogene from chromosome 11 to the immunoglobulin heavy chain gene on chromosome 14, resulting in an overexpression of cyclin D1 mRNA (also known as bcl-1 and PRAD1). In the current study performed on paraffin-embedded tissue, cyclin D1 mRNA could be detected in 23 of 24 mantle-cell lymphomas by reverse transcription polymerase chain reaction (RT-PCR) whereas only 9 of 24 demonstrated a t(11;14) by PCR. However, we also found that cyclin D1 mRNA could be detected in the majority (11 of 17, 65%) of non-mantle-cell lymphomas and in a minority of atypical lymphoid hyperplasias (3 of 7, 43%). Cyclin D1 mRNA expression was not observed in floridly reactive lymph nodes (0 of 9) or in unstimulated lymph nodes (0 of 20), suggesting that it is a sensitive adjunct marker for malignant lymphoproliferative processes, but not specific for mantle cell lymphoma. A semiquantitative RT-PCR assay was developed that compared the ratio of cyclin D1 to the constitutively expressed gene beta2-microglobulin. Using this assay on a limited number of our specimens, cyclin D1 overexpression in mantle-cell lymphoma could be reliably distinguished from its expression in other non-Hodgkin's lymphomas. This assay for cyclin D1 expression, designed for formalin-fixed, paraffin-embedded tissue, was a very sensitive and specific marker for mantle-cell lymphoma. PMID- 9846987 TI - Loss of heterozygosity and microsatellite instability in de novo versus ex adenoma carcinomas of the colorectum. AB - Small adenocarcinomas of the colorectum showing no evidence of origin from an adenoma have been called de novo carcinomas, a name that implies an origin via a different molecular genetic mechanism than the usual colorectal carcinoma which develops from an adenoma. Using microsatellite analysis, 35 early (pT1) de novo and 36 pT1 ex-adenoma carcinomas were compared using 8 microsatellite loci at 6 different chromosomal loci (1p, 2p, 8p, 5q, 17p, and 18q) known or hypothesized to be important for colorectal carcinogenesis. The rate of loss of heterozygosity (LOH) at the 17p locus (near the p53 gene) was significantly higher in the de novo than in the ex-adenoma group (73 vs. 37%, P = 0.004). The rates of LOH at the other loci (including the APC and DCC genes) and the rate of MSI were not significantly different in the two groups. These results indicate that de novo carcinomas of the colorectum develop via a similar carcinogenetic pathway as conventional ex-adenoma carcinomas; however, their higher rate of LOH at 17p is evidence for a biologically more advanced lesion with more frequent p53 mutations, consistent with clinicopathological data indicating that de novo carcinomas are more aggressive than ex-adenoma carcinomas. PMID- 9846988 TI - Expression of telomerase activity in human endometrium is localized to epithelial glandular cells and regulated in a menstrual phase-dependent manner correlated with cell proliferation. AB - Telomerase activity is observed in most malignant tumors and germ cells, whereas normal somatic cells usually do not express it. Human endometrium is composed of glandular and stromal components and exhibits dramatic changes in proliferative activity during the menstrual cycle, which is exquisitely regulated by estrogen function. We previously reported that normal human endometrium expresses telomerase activity. However, it remains unclear which of the above components are the major sources of telomerase activity and how levels of telomerase activity are regulated over the menstrual cycle. Quantitative analysis of telomerase activity revealed that it changes dramatically over the course of the menstrual cycle and is strictly regulated in a menstrual-phase-dependent manner. Maximal activity equivalent to that in endometrial cancer was present in late proliferative phase, and minimal activity in late secretory phase. Postmenopausal endometrium and endometrium treated with anti-estrogen drugs exhibited decreased telomerase activity. Testing isolated epithelial glandular cells and stromal cells, we found that telomerase activity was localized to epithelial glandular cells. In situ RNA hybridization analysis also revealed epithelial-specific expression of human telomerase RNA. In vitro analysis of cultured epithelial cells demonstrated that telomerase activity is correlated with epithelial proliferation but not affected by estrogen treatment. These findings suggest that expression of telomerase activity is specific to epithelial cells and linked to cell proliferative status. The involvement of estrogen in telomerase regulation remains to be elucidated. PMID- 9846990 TI - TRAF-4 expression in breast carcinomas. PMID- 9846989 TI - Tumor growth of FGF or VEGF transfected MCF-7 breast carcinoma cells correlates with density of specific microvessels independent of the transfected angiogenic factor. AB - We have previously shown that fibroblast growth factor (FGF)-1-, FGF-4-, or vascular endothelial growth factor (VEGF/VPF)-transfected MCF-7 breast carcinoma cells growing as tumors in nude mice are tamoxifen resistant and/or estrogen independent. These transfectants provide opportunity for study of in situ tumor induced angiogenesis promoted by the individual angiogenic factors under growth promoting versus growth-inhibiting hormonal conditions. In the present study, vessels in tumors harvested at varying times after tumor cell injection were immunohistochemically highlighted and vessel morphology and topography were scored on a scale of 0 to 4 by blinded observers. In tumors produced by all cell lines under all growth-promoting hormonal conditions, there was significantly increased abundance (P < 0.05) of edge-associated and intratumor microvessels, but not of stromally located microvessels, when compared with tumor nodules harvested under growth-inhibiting conditions, regardless of the identity of the angiogenic factor or the hormonal treatment. Image analysis of bromodeoxyuridine (BrdU)-labeled nuclei of tumors produced by all cell lines under all hormonal conditions harvested at early time points showed that mean labeling indices were highest for hormonal conditions that produced the most robust growth in that particular cell line, implying that a high BrdU labeling index is a predictor of future tumor growth in individual tumors. These results confirm previous studies that established the importance of neovascularization for tumor growth and provide validation for use of these cell lines to study the process of angiogenesis in vivo. Study of gene expression in endothelial cells in edge associated or intratumor vessels using this model might reveal mechanisms important in tumor-induced angiogenesis in human breast cancer. PMID- 9846991 TI - Interstitial cells of Cajal: pacemaker cells? PMID- 9846992 TI - Cellular and subcellular heterogeneity of glutathione metabolism and transport in rat kidney cells. AB - Selective permeabilization of plasma membranes with digitonin produced separation of cytosolic and mitochondrial compartments of proximal tubular (PT) and distal tubular (DT) cells from a rat kidney. Subcellular distributions of several intracellular glutathione (GSH)-dependent enzymes were similar in the two cell types but specific activities were significantly higher in PT cells, indicating that DT cells, particularly in their mitochondrial fraction, have a diminished capacity to detoxify reactive oxygen species. To enable isolation of suspensions of mitochondria, renal cells were treated with digitonin followed by the bacterial protease nagarse and were filtered through polycarbonate membranes. Activity distributions of enzymatic markers for subcellular fractions were quantitated and uptake of GSH was studied in suspensions of PT and DT cell mitochondria. While PT cell mitochondria catalyzed rapid uptake of GSH that was inhibited by malate, indicating involvement of dicarboxylate carriers, DT cell mitochondria exhibited limited capacity for GSH uptake that was not inhibited by substrates for the two dicarboxylate carriers. This report provides the first description of methodology for the preparation of mitochondria from renal cells derived from specific nephron cell types and shows that mitochondria from DT cells have a significantly lower capacity to use GSH for detoxification and regulation of redox status. PMID- 9846993 TI - Delayed febrile effects of chlorpyrifos: is there cross-tolerance to bacterial lipopolysaccharide? AB - Oral chlorpyrifos (CHP) induces hypothermia followed by a fever that persists for several days in the rat. To understand the neuro-immune mechanisms of CHP-induced fever, we compared the tolerance and cross-tolerance between CHP and the fever elicited by lipopolysaccharide (LPS) (Escherichia coli). Female rats were administered the corn oil (CO) vehicle or CHP (10 mg/kg; p.o.) daily for 4 days while core temperature (Tc) and motor activity (MA) were monitored by telemetry. There was a reduction in Tc followed by an elevation the next day after each CHP treatment. The day after the last CHP treatment, rats were administered saline or 50 microg/kg LPS (i.p.). CHP-treated rats had a smaller LPS fever that was attributed to their elevated baseline Tc. In another study, rats were dosed with saline or LPS daily for three days. By the time of the third LPS injection there was no febrile response, indicating tolerance to LPS. Rats were then dosed with CO or CHP (10 mg/kg) 24 h after the third LPS treatment. LPS-tolerant rats displayed an accentuated hypothermic and febrile response to CHP. Plasma cholinesterase activity was unaffected by repeated LPS treatment, suggesting that the metabolism of CHP in the liver was unaffected by LPS. Overall, the neural immune mechanisms for LPS fever is distinct from that of CHP in view of marked difference in mechanisms of tolerance. PMID- 9846994 TI - Evaluation of omeprazole genotoxicity in a battery of in vitro and in vivo assays. AB - Omeprazole, a proton pump inhibitor of wide use in the treatment of gastric acid related disorders, was evaluated for its genotoxic effects in both rat and human cultured cells and in the intact rat. DNA repair synthesis, as revealed by autoradiography, was detected in primary cultures of metabolically competent rat hepatocytes exposed to concentrations ranging from 10 to 100 mg/l, but the responses cannot be considered as clearly positive. Under the same experimental conditions any significant evidence of DNA repair was absent in primary hepatocytes from two human donors. At the same concentrations a modest but dose related increase of micronucleated cells, that reached the level of statistical significance at 33 mg/l, was present in primary rat hepatocytes and in one of two human donors. In human lymphocytes exposed to subtoxic concentrations ranging from 0.78 to 12.5 mg/l a reproducible concentration dependent clastogenic effect was absent. In partially hepatectomized female rats treated with a single p.o. dose of 1000 mg/kg, the frequency of micronucleated cells was 5.2-fold higher than in controls in the liver, but only 2.0-fold higher in polychromatic erythrocytes of the bone marrow. In rats of the same sex given azoxymethane as initiator of colon carcinogenesis the oral administration for 8 successive weeks of 10 mg/kg omeprazole on alternate days increased the response to azoxymethane, as indicated by the occurrence in colon mucosa of a modest but statistically significant increase in both the average number and size of aberrant crypt foci. Taken as a whole, our results suggest that omeprazole behaves as a weak genotoxic agent for the rat liver. Reliable information about the potential genotoxic risk to humans requires further studies on primary cells from a wide number of donors. PMID- 9846995 TI - Altered cytokine (receptor) mRNA expression as a tool in immunotoxicology. AB - Molecular immunotoxicology is aimed at analysing exposure effects on the temporal expression of important immunoregulatory genes. Cytokines play key roles in the immune system and thus molecular immunotoxicology has focused on the analysis of cytokine (expression) levels. These targets offer important new avenues to explore both in terms of mechanistic understanding of immunotoxicity and in terms of developing new assays and tests for predicting the immunotoxic potential of novel compounds. Effects on cytokine levels can be analysed on two different levels, these being mRNA and protein. The choice essentially depends on the aim of the study. Proteins comprise the biological activity so they are a more direct measure than mRNA. mRNA on the other hand, measures at a specific point in time within a tissue or organ, whereas protein is measured in a body fluid, possibly as a spill-over from tissue, or in a supernatant as a summation over a culture period. mRNA levels are assayed using Northern or dot blotting that both comprise hybridisation and using reverse transcription-polymerase chain reaction (RT-PCR). Although the latter technique has both enormous sensitivity and relative ease of operation as important advantages, it requires much more effort in terms of quantitation. References to the nucleic acid sequences of human, murine, and rat cytokines and their receptors are presented (with accession numbers). Examples in which molecular techniques were successfully employed to assess immunotoxicity and (in some cases) understand mechanisms of action are also presented. PMID- 9846996 TI - Studies on the animal model of traumatic pseudoaneurysm. AB - Sixty-nine cases of pseudoaneurysm were made in 72 femoral arteries of 54 Japanese white rabbits, with a successful rate of 95.8%. Colored Doppler's ultrasonic imaging, CT, MRI, arterial angiography, anatomic dissection, and histologic observation were carried out at different times after the operation. The mechanism for the formation of pseudoaneurysm was discussed and the effects of colored Doppler's ultrasonic imaging, CT, MRI, and arterial angiography in the diagnosis of pseudoaneurysm were evaluated. PMID- 9846997 TI - Aprotinin in ischemia-reperfusion injury: flap survival and neutrophil response in a rat skin flap model. AB - Multiple drugs have been used in experimental skin flap models to reduce the effects of reperfusion ischemia. The effects of antiproteases, however, have not been studied. A skin flap ischemia reperfusion model was developed in the rat to study the effects that aprotinin, a broad-spectrum antiserine protease, would have on skin flap viability. Thirty-two male rats underwent elevation of a ventral pedicled skin flap based on the superficial inferior epigastric artery. The flaps were subjected to 10 hr of warm ischemia by clamping the neurovascular pedicle followed by reperfusion. Aprotinin or saline (control) was administered systemically via the contralateral femoral vein either before or after the ischemic insult. Full-thickness skin biopsies were obtained at 1, 8, and 24 hr into reperfusion. Biopsies were evaluated for neutrophil concentration (using a myeloperoxidase [MPO] assay) and thromboxane B2 [TxB2] content. Flap survival was calculated at 1 week using standardized photography and computer-assisted digital imaging. Aprotinin given before an ischemic insult significantly improved flap survival compared to saline controls (52.3% alive vs. 29.6%, P = 0.0132, unpaired t-test). Aprotinin given after ischemia did not significantly influence flap survival (28.8% vs. 34.4% in saline controls, P = 0.708). MPO levels in the aprotinin preischemia treatment group were significantly less at 1 and 8 hr into reperfusion, indicating decreased neutrophil numbers. No statistical difference in TxB2 levels was noted in either group at any time after reperfusion. Aprotinin significantly improves skin flap survival when given prior to but not after an ischemic insult. Aprotinin appears to lower the concentration of neutrophils in skin flaps pretreated with the drug. Reperfused skin flap levels of thromboxane B2 are unaffected by the pre- or postischemic administration of aprotinin. PMID- 9846998 TI - Rat tail replantation as a training model for microvascular procedures of digit replantation. AB - In this study we sought to evaluate the potential of rat tail replantation as a tool for very-small-vessel microvascular anastomoses. We used 10 adult Sprague Dawley rats. The tail was completely amputated 2.0-cm distal to the base of the tail. Then the tail was replanted with anastomoses of two superficial dorsal veins from both sides and one artery. All 10 replanted tails were pink, viable, and normal-appearing at all daily inspections performed from the first to the fourteenth postoperative days. This model can provide a training tool for the acquisition of superior microvascular surgical technique for the repair of very small vessels that stimulate digital replantation. PMID- 9846999 TI - Improved surgical technique for the establishment of a murine model of aortic transplantation. AB - Aortic allotransplantation is a reliable procedure to study the evolvement of chronic rejection in mice. The progressive nature of this process in mice is characterized by diffuse and concentric myointimal proliferation which is inevitably associated with variable degrees of luminal constriction. These vascular changes are comparable to those that are witnessed in organ allografts undergoing chronic rejection in humans, underscoring its utility as a model of choice for the study of the development of this lesion. Whilst improved surgical technique has resulted in markedly enhanced graft survival, the results are far from being acceptable. Realizing this limitation, we embarked on developing a modified technique for aortic transplantation which would allow for improved graft survival in mice. A bypass conduit was created by end-to-side anastomosis of a segment of the donor's thoracic aorta into the infrarenal portion of the recipient's abdominal aorta. Using this technique, the graft survival was >98% with evidence in allotransplanted aorta of morphological changes pathognomonic of chronic rejection. On the contrary, no histopathological anomalies were discerned in aortic grafts transplanted across syngeneic animals. This modified surgical approach ameliorates the unacceptably high graft loss associated with earlier techniques, further extending the utility of this model as a tool to study the molecular and cellular mechanisms rudiment to the evolvement of chronic rejection. PMID- 9847000 TI - Double free-flap reconstruction of massive defects involving the lip, chin, and mandible. AB - Two patients with massive, composite defects of the total lower lip, chin, and anterior mandible underwent double free-flap reconstruction. A fibular osteoseptocutaneous flap was used to reconstruct the mandible and floor of the mouth and a radial forearm fasciocutaneous composite flap, including the palmaris longus tendon, was used for total lower lip and chin reconstruction. Postoperatively, both patients had acceptable cosmesis, were orally competent, and recovered adequate mandibular function. Double free-flap reconstruction is indicated only in those circumstances in which composite tissue requirements or massive tissue defects preclude reconstruction with a single free-tissue transfer. PMID- 9847001 TI - Regeneration through long nerve grafts in the swine model. AB - We investigated regeneration across a long nerve defect in the swine model to study extensive neural loss and long nerve gap. Most experiments have been conducted in the rodent model that, while an appropriate immunological model, only allows short nerve gaps to be studied. Twelve outbred swine received either an 8-cm ulnar nerve autograft or an allograft without immunosuppression. At 6 and 10 months, histomorphometry of the autografts demonstrated excellent nerve regeneration, while very poor regeneration was noted across the allografts. This confirmed that 8 cm are an adequate challenge independent of the spontaneous regeneration potential of axons seen in rodents. The swine ulnar nerve graft model causes minimal morbidity and will now be used with immunological manipulation of inbred animals. PMID- 9847002 TI - Donor site morbidity following vascularized fibular grafting. AB - We examined donor site morbidity in thirty-nine patients with avascular necrosis of the femoral head treated by curettage and transplantation of a free ipsilateral fibular graft. Utilising our donor site morbidity questionnaire, scar, functional loss, wound healing, complications, and pain were analysed. Subjective complaints and objective findings were evaluated and compared. Subjective complaints were common and included a sense of instability in 42% and a sense of weakness in 37%. However, objective findings were limited. No clinical instability could be elicited and only great toe flexion (29%) and extension (43%) were found to be mildly weak. Only one patient required reoperation for a donor site problem (2%). Eighty-nine percent were pain free at time of follow-up, and 93% felt the scar was good. Range of motion of the knee and ankle of the donor site leg was not different from the nonoperated leg. Donor site morbidity for avascular necrosis of the femoral head is low. PMID- 9847003 TI - Heat-induced tissue fusion for microvascular anastomosis. PMID- 9847004 TI - Determination of common drugs of abuse in body fluids using one isolation procedure and liquid chromatography--atmospheric-pressure chemical-ionization mass spectromery. AB - A method for determining opiate agonists (morphine, morphine-3-glucuronide, morphine-6-glucuronide, 6-monoacetylmorphine, codeine, codeine-6-glucuronide, dihydrocodeine, dihydromorphine, buprenorphine, methadone, tramadol, and ibogaine), cocaine and its metabolites (benzoylecgonine and ecgonine methyl ester) and lysergic acid diethylamide in serum, blood, urine and other biological matrices is presented. Aliquots (0.5-1.5 mL) of biological fluids were spiked with appropriate deuterated internal standards and extracted using a common solid phase extraction method (C18 cartridges). The extracts were subjected to liquid chromatographic-atmospheric-pressure chemical-ionization mass spectrometric examination using selected ion monitoring procedures. These procedures were developed after analysis of full-scan mass spectra of examined compounds. The extraction method appeared very universal; the recoveries were high for almost all drugs and the extracts were very clean. The procedure was applied for routine forensic casework. PMID- 9847005 TI - Benzodiazepines: toxic serum concentrations in positive enzyme immunoassay responses. AB - A total of 588 blood specimens collected in an emergency unit were screened for benzodiazepines (BZDs) using enzyme-multiplied immunoassay and gas chromatography. Two-hundred eighty-five samples were positive for BZDs, and 303 samples that were negative by EMIT included 20 samples with BZDs detectable by gas-liquid chromatography. A total of 15 BZDs were identified, and the most frequently occurring were nordiazepam, bromazepam, diazepam, and alprazolam. Individual BZDs were found in 74% of cases, but some samples contained two, three, or even four BZDs. There is a risk of missing intoxication by BZDs with low therapeutic range and/or low cross-reactivity (alprazolam, bromazepam, flunitrazepam). There is a risk of misinterpreting a positive result for some BZDs with high therapeutic range and/or high cross-reactivity (nordiazepam), which may reflect a pharmacologically ineffective concentration. A semiquantitative analysis is inappropriate even when the identity of BZD is known. Immunoassays are the only methods presently available for use in emergencies, but physicians must be clearly informed of their limitations and interpret results with caution. PMID- 9847006 TI - In vivo formation of codeinone-glutathione adduct: isolation and identification of a new metabolite in the bile of codeine-treated guinea pig. AB - Codeinone-glutathione adduct (CO-GSH) in the bile of guinea pigs given a subcutaneous injection of codeine was isolated and identified. Synthesized authentic CO-GSH was characterized by the mass and nuclear magnetic resonance spectra and used as the standard sample. The metabolite was isolated by preparative high-performance liquid chromatography on a C18 column. The fractions containing the conjugated metabolite were purified using Sep-Pak C18 cartridges. For further purification of the metabolite CO-GSH, a Radial Pak CN column was used. Structure assignment of the metabolite was then performed by fast-atom bombardment mass spectrometry and 500 MHz Fourier-transform-NMR spectrometric analysis and identified as S-[4,5-epoxy-3-methoxy-17-methyl-6-oxomorphinan-(8S) yl] glutathione. PMID- 9847007 TI - Validation of an automated microplate enzyme immunoassay for screening of postmortem blood for drugs of abuse. AB - The objective of this study was to compare the sensitivity and specificity of an enzyme immunoassay employing antibodies bound to a microtiter plate (MPEIA) with those of two radioimmunoassays for screening postmortem blood from selected coroner's cases for drugs of abuse. The radioimmunoassays were a coated-tube radioimmunoassay (CTRIA) and a double antibody radioimmunoassay (DARIA). Specimens consisted of 260 postmortem blood specimens from coroner's cases. Immunoassay results (positive or negative) were compared with confirmed results on those cases by gas chromatography-mass spectrometry, alone or in combination with gas-liquid chromatography using either a nitrogen-phosphorus or flame ionization detector. Sensitivity was calculated as the true-positive rate using chromatographic confirmation as the reference standard. Specificity was calculated as the true-negative rate. Sensitivity and specificity were calculated for 5-7 potential cutoff concentrations for the drug classes opiates, amphetamines, cocaine and metabolites, and barbiturates. For opiates, the sensitivity and specificity were 99% and 93%, respectively, for the MPEIA at a cutoff of 20-ng/mL morphine, compared with 94% and 96% for the CTRIA at a cutoff of 5-ng/mL morphine and > 99% and 96% for the DARIA at 20-ng/mL morphine. For cocaine and metabolites, the sensitivity and specificity were 96% and 93%, respectively, for the MPEIA at 50-ng/mL benzoylecgonine, compared with 93% and 96% for CTRIA at 50-ng/mL benzoylecgonine and 98% and 97% for the DARIA at 50 ng/mL benzoylecgonine. For amphetamines, the sensitivity and specificity were >99% and 91%, respectively, for the MPEIA at 25-ng/mL methamphetamine, compared with 93% and 86% for the CTRIA at 25-ng/mL methamphetamine and 83% and 89% for the DARIA at 50-ng/mL methamphetamine. For barbiturates, the sensitivity and specificity were > 99% and 92%, respectively, for the MPEIA at 50-ng/mL secobarbital, compared with 91% and 87% for the CTRIA at 500-ng/mL secobarbital and 79% and 95% for the DARIA at a cutoff of 1000-ng/mL phenobarbital. PMID- 9847008 TI - Occurrence of cocaine in urine of substance-abuse treatment patients. AB - As part of ongoing research efforts to improve methods of monitoring drug use in treatment patients, the presence of cocaine in urine specimens was evaluated as a possible marker for recent illicit cocaine use. A total of 2327 urine specimens collected during a 17-week clinical trial of a cocaine-abuse treatment study were tested. Cocaine was measured by gas chromatography-mass spectrometry, and benzoylecgonine (BZE) equivalents were determined by fluorescence polarization immunoassay (FPIA). More than one-third of the specimens were positive (> 25 ng/mL) for cocaine (36.8%), and nearly two-thirds were positive (> 300 ng/mL) for cocaine metabolite by FPIA (62.7%). Median concentrations of cocaine and BZE equivalents were 235 and 14,900 ng/mL, respectively, and maximum concentrations were 112,025 and 1,101,190 ng/mL in cocaine- and BZE-positive specimens, respectively. There were 52 specimens that contained cocaine in equal or higher concentrations than BZE equivalents. No significant differences in cocaine or BZE concentrations between Caucasian and African-American or between male and female patients were found. Cocaine was present less frequently and at lower concentrations than BZE but more frequently than expected based on an average half-life of approximately 1 h, which suggests that cocaine may exhibit a longer terminal half-life and/or that accumulation of cocaine can occur in chronic, heavy users. PMID- 9847009 TI - Relative binding of therapeutic drugs by sera of seven mammalian species. AB - The relative binding of acetaminophen, lidocaine, phenobarbital, procainamide, quinidine, and theophylline to sera of seven mammalian species was studied. Pooled commercial sera from cow, goat, horse, human, pig, rabbit, and sheep were supplemented with 5 and 10 mM concentrations of each drug. For each serum, each drug, and each drug concentration, equilibrium dialysis was performed in duplicate against phosphate buffer (pH 7.4, 0.1 M, 4 degrees C). Percent drug bound to serum was calculated. Phenobarbital demonstrated more than 20% binding to goat, horse, human, and sheep serum at both 5 and 10 mM concentrations; more than 20% binding to bovine serum at a concentration of 10 mM; and more than 20% binding to pig and rabbit serum at 5 mM. Quinidine (studied only at 5mM concentration) bound more than 20% to cow, goat, horse, human, pig, and rabbit serum. In contrast, procainamide at both the 5 and 10 mM concentrations showed no binding to cow, horse, pig, rabbit, or sheep serum. Acetaminophen (studied only at 5 mM concentration), lidocaine, and theophylline demonstrated less than 20% binding to each serum. Acetaminophen at 5 mM did not bind to human serum, and lidocaine at 10 mM did not bind to horse or pig serum. Although some interspecies variation in drug binding to the seven sera was noted, the overall magnitude of binding of each drug to each serum was, for the most part, similar. Phenobarbital and quinidine showed stronger (> 20%) binding; procainamide showed negligible binding; and acetaminophen, lidocaine, and theophylline demonstrated intermediate (< 20%) binding. PMID- 9847010 TI - Determination of bromvalerylurea and its metabolites in biological samples by frit-fast atom bombardment liquid chromatography-mass spectrometry. AB - A simple, rapid, and sensitive method which allows us to simultaneously determine bromvalerylurea (BVU) and its three metabolites (3-methylbutyrylurea [MVU], alpha (cystein-S-yl)isovalerylurea [CVU], and alpha-(N-acetylcystein-S yl)isovalerylurea [AcCVU]) was investigated by frit-fast atom bombardment liquid chromatography-mass spectrometry (frit-FAB LC-MS). The LC-MS analysis was performed after the solid-phase extraction from tissue and urine samples with a Sep-Pak C18 cartridge. Tissue homogenates and urine were adjusted to pH 4.0 and applied to the cartridges. The retained BVU and its metabolites were eluted from the cartridge with 2 mL of acetonitrile/10 mM ammonium acetate buffer (pH 3.5, 50:50, v/v). The eluate was analyzed by LC-MS, which employs a semimicro type L column ODS column. The proposed conditions are as follows: mobile phase A, 0.4% glycerol in acetonitrile/10 mM ammonium acetate buffer (pH 3.5) (5:95, v/v); mobile phase B, 0.4% glycerol in acetonitrile; elution mode, linear gradient, 100% A (5 min) to 100% B in 15 min; flow rate, 0.2 mL/min; split ratio, 1:40. Extraction recoveries of BVU and its metabolites were 91.90-97.79% from the spiked liver homogenate and 89.68-96.13% from the spiked urine. The detection limits ranged from 10 to 25 ng/g in selected ion monitoring mode. PMID- 9847011 TI - Caffeine demethylation monitoring using a transdermal sweat patch. AB - Caffeine and two metabolites (paraxanthine and theobromine) were quantitated by high-performance liquid chromatography (HPLC) using extracts from transdermal sweat patches that continuously collected and stored analytes lost through the skin. Following caffeine consumption, remarkably clean chromatograms were obtained with minimal sample preparation. Caffeine and paraxanthine accumulated in the patch at comparable rates, and theobromine accumulated more slowly. A major urinary metabolite, 1-methylxanthine, was notably absent in sweat-patch and plasma extracts, a finding which favors a renal source for this metabolite. A simple, noninvasive approximation of N-demethylation can be made by calculating the paraxanthine/ caffeine and theobromine/caffeine ratios in the patch extract. These ratios were significantly reduced in high-dose (600 mg) versus low-dose (200 mg) subjects, possibly reflecting a decreased clearance of caffeine. Because the sweat patches can be worn for several days, the technique gives a multiday historical record which reflects the fluctuating systemic concentration of caffeine and its hepatic metabolites and thus might be useful to noninvasively monitor compliance by caffeine-restricted patients or to assess drug-metabolizing status. PMID- 9847012 TI - Evaluation of the diagnostic performance of the Boehringer Mannheim CEDIA LSD assay. AB - The precision and the diagnostic performance of the Boehringer Mannheim CEDIA DAU LSD assay was evaluated. The assay was performed in the semi-quantitative mode on a Hitachi 917 analyzer. Within-run coefficients of variation (CVs) of the semiquantitative values for 0.25 and 1.0 ng/mL were 11.2 and 6.2%, respectively. Day-to-day CVs for the same concentrations were 12.6 and 8.6%. We analyzed 318 urine samples by CEDIA, DPC Coat-A-Count RIA and Behring EMIT II. Confirmation was performed by GC-MS, after extraction on Bond Elut Certify columns. Two hundred sixty-three samples were negative by all methods. Twenty-five samples were positive by all immunoassays, 19 of which were confirmed by gas chromatography-mass spectrometry (GC-MS). One sample was falsely negative by CEDIA. Three samples were positive by EMIT and CEDIA, but negative by RIA and GC MS. Twenty-six samples were positive by EMIT alone, but they were not confirmed by GC-MS. The LSD CEDIA assay seems to be less specific than DPC RIA but more specific than the EMIT LSD assay. PMID- 9847014 TI - Organochlorine pollutants in human hair. PMID- 9847013 TI - Determination of olanzapine in a postmortem case. AB - Olanzapine, a new antipsychotic agent, was approved by the U.S. Food and Drug Administration in 1996 for use in the treatment of schizophrenia. It is structurally similar to clozapine, has a low incidence of extrapyramidal effects, and is effective in treating both the positive and negative symptoms of schizophrenia. This paper describes the determination of olanzapine in biological specimens obtained from the autopsy of a 35-year-old white male found dead in bed at a psychiatric facility. In the months prior to his death, the deceased was prescribed multiple medications, including olanzapine. Olanzapine was identified qualitatively by full scan gas chromatography-mass spectrometry, with quantitative analysis performed by liquid-liquid extraction followed by dual column gas chromatography. The following concentrations were determined in the specimens analyzed: heart blood, 550 ng/mL; bile, 6346 ng/mL; and gastric contents, 157 ng/mL. Vitreous humor, cerebrospinal fluid, and urine specimens were negative. Although steady-state plasma concentrations of 10-25 ng/mL olanzapine have been reported, effective levels are known to be highly variable and a plasma concentration of 300 ng/mL has been tolerated without adverse effects. Based upon the autopsy, toxicological findings, and case investigation, the cause of death was determined to be intramyocardial arteriosclerosis with severe stenosis of the nodal artery due to hypertensive cardiovascular disease, and the manner was natural. PMID- 9847015 TI - Plasma concentrations after a clomipramine intoxication. PMID- 9847016 TI - Human interleukin 10 suppresses production of inflammatory mediators by LPS stimulated equine peritoneal macrophages. AB - To investigate the ability of recombinant human interleukin 10 (rhuIL-10) to suppress the release of inflammatory mediators from lipopolysaccharide (LPS) stimulated equine macrophages, rhuIL-10 was added to equine peritoneal macrophage monolayers at concentrations of 0, 0.1, 1, 10, or 100 ng/ml. Thirty minutes later, LPS (E. coli O55:B5) was added at final concentrations of 0, 1, 10, 100 ng/ml. Macrophages were incubated for 16 h at 37 degrees C, then supernates were harvested and assayed for tumor necrosis factor (TNF) activity (L929 cytotoxicity), interleukin-6 (IL-6) activity (B9 proliferation), prostaglandin E2 concentration (ELISA), and nitric oxide (Griess reaction for nitrite). Preincubation of LPS-stimulated peritoneal macrophages with rhuIL-10 caused significant (P<0.05) reduction in secretion of TNF, IL-6, and PGE2, in a dose dependent manner. Of the inflammatory mediators, TNF was most sensitive to the effects of rhuIL-10. At concentrations of rhuIL-10> or =1 ng/ml, TNF activity in the supernate was inhibited significantly at all concentrations of LPS. At one or more LPS concentrations, there was significant inhibition of each mediator in the presence of 1 ng rhuIL-10/ml and, at 100 ng/ml, rhuIL-10 significantly inhibited production of each mediator at all LPS concentrations tested. When data were expressed as a percentage of control values and pooled across all LPS concentrations, both PGE2 and TNF values were significantly reduced at rhuIL-10 concentrations of > or =1 ng/ml, whereas IL-6 was inhibited significantly at concentrations of > or =10 ng rhuIL-10/ml. Tumor necrosis factor production was more completely suppressed (7.8% of control) by the highest concentration of rhuIL-10(100 ng/ml) than was PGE2 (27.2%) or IL-6 (43.8%). Nitrite was not detected in any supernate from peritoneal macrophage monolayers. PMID- 9847017 TI - Dose-dependent priming or desensitization induced by chemotactic agents in chemiluminescence experiments with canine and human neutrophils. AB - We explored whether receptor-specific desensitization in repeated stimulation offers an opportunity to identify chemotactic factors particularly in species, e.g. the dog, for which immunochemical methods to determine chemotactic factors are not commercially available. Complement fragment 5a and interleukin-8 act via distinct receptors. They were used as test agents for neutrophils in luminol dependent chemiluminescence. These experiments led to the observation that exposure of human and canine neutrophils to low concentrations of a chemotactic agent shows an increased response when stimulated again with the same agent (priming). In the heterologous system even cross-priming could be observed after a second stimulation with another agent. The concentration at which priming rather than desensitization occurred were lower for homologous factor-cell combinations (<10-20 nM) than for heterologous combinations (<45-60 nM). The mechanism underlying this phenomenon is unknown. However, it raises the possibility that it subserves an important function in the recruitment and activation of cells by low agonist concentration. This study shows that despite the phenomenon of priming receptor-specific desensitization may be used to determine chemotactic agents in the homologous system. PMID- 9847018 TI - Evaluation of GM1 ganglioside-mediated apoptosis in feline thymocytes. AB - Cats with inherited GM1 gangliosidosis (GM1 mutant cats) have premature thymic involution characterized by decreased total thymocytes primarily affecting the CD4+ CD8+ subpopulation. While GM1 mutant cats have increased cell surface GM1 gangliosides, as determined by cholera toxin B binding, on both thymocytes and peripheral lymph node cells only thymocytes show increased apoptosis. To determine if GM1 gangliosides can increase the occurrence of apoptosis in feline thymocytes directly, we added exogenous GM1 ganglioside (GM1) to feline thymocyte primary cultures and compared the results to apoptotic changes seen in untreated cells or in cells treated with dexamethasone (Dex), a known inducer of thymocyte apoptosis in other species. Incorporation of exogenous GM1 into thymocyte cytoplasmic membranes was confirmed by flow cytometric analyses of cholera toxin B labelling. Apoptosis in feline thymocytes was analyzed by electron microscopy, spectrophotometric evaluation of DNA fragmentation, flow cytometric enumeration of apoptotic nuclei, and gel electrophoretic analysis of degraded DNA. Alterations in percentages of thymocyte immunophenotype following GM1 incorporation were determined by flow cytometric analyses of labelled cell surface markers for feline CD4 and CD8. Because in vitro addition of GM1 gangliosides has been reported in other species to decrease surface expression of CD4 on both thymocytes and peripheral lymphocytes, we evaluated GM1-associated down-regulation of CD4 on the surface of feline thymocytes and peripheral lymph node cells by flow cytometry. Additionally, we compared the apoptotic response of the more mature peripheral lymph node cells to the less mature thymocytes. Our results indicate that incorporation of exogenous GM1 into feline thymocyte cell membranes produces a dose-dependent increase of apoptotic cell death. Although, CD4 expression on both feline thymocyte and lymph node cell membranes was abruptly decreased after introducing exogenous GM1, enhanced apoptotic death was observed only in thymocytes, not in lymph node cells at the same GM1 concentration. Enhancement of thymocyte apoptosis appears to be age-related since cells derived from cats <3 months of age were more vulnerable than those from cats >3 months of age. PMID- 9847019 TI - Comparative study of histamine release from skin mast cells dispersed from atopic, ascaris-sensitive and healthy dogs. AB - Atopic dermatitis results from the interaction between allergen and allergen specific IgE bound to the mast cell surface receptors. This process triggers mast cell degranulation and accounts at least for early phase reaction. Furthermore, there is increasing in vitro and in vivo evidence that IgE has the ability to induce overexpression of the Fc epsilonRI receptor on the mast cell plasma membrane. In order to study the potential effect of an increase in serum IgE on mast cell activity, the histamine releasability of mature mast cells isolated from the skin of atopic, ascaris-sensitive and healthy dogs was analyzed. No histamine release was detected upon the immunological stimulation of cells that were not previously sensitized with atopic or ascaris-sensitive dog serum. However, when passively sensitized, skin mast cells were challenged with either Asc SI antigen or anti-IgE, the mast cell histamine release increased in a stimulus concentration-dependent manner. The amount of histamine released was significantly higher in response to anti-IgE than in response to Asc SI antigen. However. the difference in the percentage of mast cell histamine release between atopic (26.3+/-2.8%) and non-atopic (30.9+/-1.7%) dogs was not statistically significant, similar to what occurred when ascaris-sensitive (12.8+/-1.6%) and non-sensitive (13.2+/-1.7%) dogs were compared. Although these results could suggest that there is either little or no increase in the density of IgE receptors in atopic or ascaris-hypersensitive dogs versus controls, we strongly consider either the possibility that the digestion procedure might affect cell behaviour in vitro or that an underlying increase of receptors poorly affects the release of granule-stored mediators but influences mast cell activity in a different manner. PMID- 9847020 TI - The role of neutrophil chemotactic cytokines in the pathogenesis of equine chronic obstructive pulmonary disease (COPD). AB - Chronic obstructive pulmonary disease (COPD) is defined as a chronic obstructive inflammatory disease affecting the small airways associated with hay dust exposure (Lowell, F.C., 1964. Observation on heaves. An asthma like syndrome in the horse, J. Allergy 35, 322-330). The disease corresponds histopathologically to a chronic bronchiolitis (Gerber, H., 1973. Chronic pulmonary disease in the horse, Equine Vet. J. 5, 26-33; Winder, N.C., Grunig, G., Hermann, M., Howald, B., von Fellenberg, R., 1989. Comparison of respiratory secretion cytology and pulmonary histology in horses, J. Vet. Med., A36, 32-38) and is mainly characterized by the presence of neutrophil granulocytes in the small bronchioles. Around 12-50% of all horses in Europe and the northern United States suffer from this disease (Mc Pherson, E.A., Lawson, G.H.K., Murphy, J.R., Nicholson, J.M., Fraser, J.A., Breeze, R.G., Pirie, H.M., 1978. Chronic obstructive pulmonary disease (COPD): Identification of affected horses, Eq. Vet. J. 10, 47-53; Larson, V.L., Busch, R.H., 1985. Equine tracheobronchial lavage: Comparison of lavage cytologic features in horses with chronic obstructive pulmonary disease, Am. J. Vet. Res., 46, 144-146; Bracher, V., von Fellenberg, R., Winder, N.C., Grunig, G., 1991. An investigation of the incidence of chronic obstructive pulmonary disease (COPD) in random populations of swiss horses, Equine Vet. J. 23, 136-141). The number of neutrophils in the bronchoalveolar lavage (BAL) and in tracheobronchial secretions (TBS) correlates with the severity of the disease. The present study is focused on the mechanisms which lead to the infiltration of neutrophil granulocytes in the lung of horses. We found that: (1). A strong chemotactic activity in the BAL fluid is associated with high levels of dust exposition. (2). In vitro stimulated alveolar macrophages have impaired phagocytosis efficiency and secrete two chemo attractants specific for neutrophil granulocytes: Interleukin-8 (IL-8) (Wuyts, A., Proost, P., Put, W., Lenaerts, J.-P., Paemen, L., van Damme, J., 1994. Leucocyte recruitment by monocyte chemotactic proteins (MCPs) secreted by human phagocytes, J. Immunol. Meth. 174, 237-247) and macrophage inflammatory protein-2 (MIP-2) (Wolpe, S.D., Sherry, B., Juers, D., Davatelis, G. Yurt, R.W., Cerami, A., Identification and characterisation of macrophage inflammatory protein-2, Proc. Natl. Acad. Sci. USA 86, 612-616; Tekamp-Olson, P., Gallegos, C., Bauer, D., 1990. Cloning and characterisation of cDNAs for murine macrophage inflammatory protein-2 and its human homologues, J. Exp. Med., 172, 911-927; Driscoll, K.E., 1994. Macrophage inflammatory proteins: Biology and role in pulmonary inflammation. Exp. Lung Res., 20, 473-490). This is associated with the appearance of chemotactic activity in the supernatant. These data confirmed our working hypothesis that bronchiolar neutrophilia may be the consequence of a (over)stimulation of pulmonary macrophages leading to expression of cytokines chemotactic for neutrophil granulocytes. PMID- 9847021 TI - Phenotype, growth regulation and cytokine transcription in Ovine Herpesvirus-2 (OHV-2)-infected bovine T-cell lines. AB - The causal agent of sheep-associated malignant catarrhal fever (MCF), Ovine Herpesvirus-2 (OHV-2), can be propagated in IL-2-dependent lymphoblastoid cell lines derived from diseased cattle and deer providing a useful model for the investigation of the pathogenesis of MCF. In this study, five interleukin-2 (IL 2)-dependent cell lines were established from affected cattle to examine their growth regulation and cytokine transcription. All cell lines expressed CD2, CD5 and CD25. Three of the cell lines were CD4+ and one CD8+, whereas one cell was of mixed CD4 and CD8 phenotye. The growth of these cell lines was reduced when cultured with antibody against CD25, the IL-2 receptor alpha subunit. All cell lines showed a lack of response to Con A and their cell growth was inhibited by Cyclosporin A which is known to inhibit cytokine promoters. It was decided therefore, to examine the cell lines for the presence of mRNA of different cytokines. The results showed that the cell lines transcribed message for IFNgamma, TNFalpha, IL-4 and IL-10 whereas no mRNA for IL-2 or IL-1beta was detected. In conclusion, the OHV-2-immortalised cell lines resemble anergic T cells which may be activated giving rise to the characteristic lesions of MCF. PMID- 9847022 TI - Stress enhanced reduction in peripheral blood lymphocyte numbers in dairy cows during endotoxin-induced mastitis. AB - Two groups of Holstein dairy cows, which showed either high- (HC; n = 8) or low (LC; n = 10) plasma cortisol concentrations in response to a psychological stressor (novel-environment test) in their first lactation, were used 1 year later for studying the effects of psychological stress on various aspects of the host defence during endotoxin-induced mastitis. Social isolation was used as a stressor; endotoxin was used to activate the host defence. HC-cows appeared to be more stressed by the experimental procedure than LC-cows. One hour after having been isolated, rectal temperature increased (+0.59 degrees C) in HC- but not in LC-cows. Handling, related to the intra-mammary administration of endotoxin, induced a higher increase in cortisol concentration in HC- than in LC-cows 1 h after injection. In addition, HC-cows vocalised significantly more frequently (595 +/- 222) than LC-cows (81 +/- 24) between 1.5 and 9.5 h after isolation. Between 3 and 5.5 h after endotoxin-treatment, cortisol concentrations increased profoundly in both LC- and HC-cows and peaked at 24.6 +/- 3.8 and 22.8 +/- 3.1 ng ml(-1), respectively. The rise in plasma cortisol concentration was associated with a decrease in the number of circulating lymphocytes between 0 and 8 h post injection (PI). Between 8 and 10 h PI, the number of circulating lymphocytes in HC-cows further decreased (40%) to 1.58x10(6) cells ml(-1) and remained so until 21 h PI. This was not the case with the LC-cows. The results show that a differing sensitivity of dairy cows to environmental stressors is associated with a significant difference in the number of circulating leukocytes following intra mammary administration of endotoxin during a period of social isolation. The results further suggest that, apart from adrenocortical hormones, other factors related to general stress-responsiveness modulate the number of peripheral lymphocytes in dairy cows during endotoxin-induced mastitis. During environmental stress, these factors may enhance endotoxin-induced reduction in circulating lymphocyte numbers, particularly in cows that are sensitive to environmental stressors. As to the clinical relevance of these differences in stress responsiveness, no differences between LC- and HC-cows were found in disease incidence nor in duration of a disease episode during both the 1st and 2nd lactation. In the 2nd lactation. HC-cows tended to produce less milk than LC cows. PMID- 9847023 TI - Signs, symptoms, and ill-defined conditions in Persian Gulf War veterans: findings from the Comprehensive Clinical Evaluation Program. AB - OBJECTIVE: The purpose of this study was to analyze the type and frequency of signs, symptoms, and ill-defined conditions (SSID; International Classification of Diseases-9th Revision, Clinical Modification (ICD-9-CM) codes 780-799) identified by physicians evaluating Persian Gulf War veterans; to determine the influence of the extent of evaluation on the type and frequency of SSID diagnoses; and to search for evidence for a new illness, or illness related to wartime exposures, in veterans with ill-defined conditions. METHOD: Comprehensive examinations were provided for 21,579 consecutive Persian Gulf War veterans with symptoms or health concerns after the war. Data recorded on all individuals includes demographics, self-reported exposures, symptoms, and physician-assigned ICD-9-CM primary and secondary diagnoses. A detailed psychosocial history, including a multidisciplinary discussion, was incorporated for a subset of participants. RESULTS: SSID conditions were primary diagnoses for 17.2% of veterans, and either primary or secondary diagnoses for 41.8%. Although some SSIDs were objective conditions (eg, sleep apnea), most were simply symptoms. More comprehensive evaluation, especially the multidisciplinary discussion of findings, decreased the frequency of symptoms as diagnoses and increased the number of DSM-IV psychiatric diagnoses. Ill-defined conditions were not associated with particular self-reported exposures or demographic variables. CONCLUSIONS: Ill-defined conditions identified by physicians in Gulf War veterans are most often symptoms. More definitive, often psychological, diagnoses can be made by increasing the intensity of the evaluation and by multidisciplinary input. Evidence for a new or unique illness related to wartime exposures did not emerge from this analysis. PMID- 9847024 TI - Identifying illness: pitfalls and discoveries. PMID- 9847025 TI - Voluntary breath holding in panic and generalized anxiety disorders. AB - OBJECTIVE: Because breath holding causes arterial pCO2 to increase, we used it to test the hypothesis that in panic disorder (PD) a biological suffocation monitor is pathologically sensitive. METHOD: Nineteen patients with PD, 17 with generalized anxiety disorder (GAD), and 22 normal controls took deep breaths on signal and held them until a release signal was given 30 seconds later. This was repeated 12 times separated by 60-second normal breathing periods. RESULTS: PD patients reported having had in the past more symptoms of shortness of breath when anxious, and more frequent frightening suffocation experiences than the other groups. However, increases in self-rated anxiety between periods of normal breathing and periods of breath holding were similar in all three groups. Skin conductance, blood pressure, and T-wave amplitude reactions to breath holdings were also similar, but heart rate acceleration upon taking a deep breath was greater in GAD patients. Before and after individual breath holdings, end-tidal pCO2 was lower in PD patients than in normal controls; GAD patients were intermediate. Inspiratory flow rate did not differ between groups. CONCLUSIONS: Our physiological results provide no direct support for an overly sensitive suffocation alarm system in PD. Lower pCO2 may be due to anxiety causing hyperventilation in patients prone to panic. PMID- 9847026 TI - Breathlessness, anxiety, and respiratory physiology. PMID- 9847027 TI - An historical and critical review of the articles on blood pressure published in Psychosomatic Medicine between 1939 and 1997. AB - Between 1939 and 1997, there have been 59 volumes of Psychosomatic Medicine. Over this period there were 200 articles dealing with blood pressure. About 90% of these were concerned with high blood pressure. This article reviews all of these papers both from an historical perspective and critically. Although there has been a significant growth in the rate of articles published since 1939, there has always been a strong interest in the nature of hypertension, particularly the roles of affects and emotions in the natural history of the disease. For example, volume 1, number 1 of the Journal includes a symposium on high blood pressure in which Franz Alexander stated his well-known hypothesis that the chronic inhibition of rage plays a causal role in the production of hypertension. In various forms, the notion that anger is an important mediator of hypertension has neither been proved nor abandoned. One major conclusion drawn from this review is that the current research on high blood pressure is drifting somewhat aimlessly. It has become preoccupied with demonstrations that various stimuli or situations (usually characterized as stresses) can acutely raise blood pressure. Despite this focus, neither the necessary nor the sufficient conditions for labeling a stimulus as stress has ever been agreed upon. Likewise, there have been many demonstrations of an iatrogenic effect on blood pressure, but neither the behavioral mechanisms underlying this effect nor the strategies for eliminating it have been explicated. Finally, this article identifies several areas where it would be useful to review and integrate current knowledge. Hopefully, such integrations could play a significant role in focusing and shaping future research and clinical practice. PMID- 9847028 TI - A longitudinal study of job strain and ambulatory blood pressure: results from a three-year follow-up. AB - OBJECTIVE: This study was conducted to investigate the hypothesis that exposure to "job strain" is related to increased ambulatory blood pressure (ABP). METHOD: Participants were 195 men who wore an ABP monitor for 24 hours on two occasions 3 years apart. Job strain status, evaluated at each assessment, was used to identify four groups: those not having job strain at either assessment (N = 138), those having job strain at both times (N = 15), and two crossover groups. Regression analysis was used to examine the cross-sectional associations of ABP with job strain, controlling for other known risk factors. Repeated-measures analysis of variance was used to examine the association of ABP change with the category of job strain change. RESULTS: The cross-sectional analysis of the Time 2 data yielded almost identical, highly significant effects of job strain on ABP as was shown in our previously published Time 1 analysis. Those in high strain jobs at both times had systolic/diastolic ABPs at work and at home that were, on average, 11/7 mm Hg higher than those with no job strain at both times; the crossover groups had intermediate levels of ABP. The longitudinal analysis showed that those with a high strain job at Time 1, but not at Time 2, had a significant decrease in work and home ABP of approximately 5/3 mm Hg. CONCLUSIONS: The previously reported cross-sectional association between job strain and ABP was replicated at follow-up. The group repeatedly exposed to job strain had higher levels of ABP at Time 2 than either crossover group. Furthermore, change in job strain status partially predicted change in ABP. These results provide new evidence supporting the hypothesis that job strain is an occupational risk factor in the etiology of essential hypertension. PMID- 9847029 TI - Social support and hostility as predictors of depressive symptoms in cardiac patients one month after hospitalization: a prospective study. AB - OBJECTIVE: Hospitalization for cardiac disease is associated with an increased risk for depression, which itself confers a poorer prognosis. Few prospective studies have examined the determinants of depression after hospitalization in cardiac patients, and even fewer have examined depression within the weeks after hospital discharge. The present study assessed the prospective relations among perceptions of social support and trait hostility in predicting symptoms of depressive symptoms at 1 month after hospitalization for a diagnostic angiography in 506 coronary artery disease (CAD) patients. METHOD: A series of structural equation models 1) estimated the predictive relations of social support, hostility, and depressive symptoms while in the hospital to symptoms of depression 1 month after hospitalization, and 2) compared these relations across gender, predicted risk classification, and age. RESULTS: Social support assessed during hospitalization was independently negatively associated with depressive symptoms 1 month after hospitalization, after controlling for baseline symptoms of depression, gender, disease severity, and age. Hostility was an indirect predictor of postdischarge depressive symptomology by way of its negative relation with social support. This pattern of relations did not differ across gender, predicted risk classification, and age. CONCLUSIONS: Our findings suggest that a patient's perceived social support during hospitalization is a determinant of depressive symptoms 1 month later. The relation of social support and hostility to subsequent depressive symptoms was similar across a variety of populations. PMID- 9847030 TI - Stressful events, pessimism, natural killer cell cytotoxicity, and cytotoxic/suppressor T cells in HIV+ black women at risk for cervical cancer. AB - OBJECTIVE: This study examines whether stressful negative life events and pessimism were associated with lower natural killer cell cytotoxicity (NKCC) and T cytotoxic/suppressor cell (CD8+CD3+) percentage in black women co-infected with human immunodeficiency virus Type 1 (HIV-1) and human papillomavirus (HPV), a viral initiator of cervical cancer. METHOD: Psychosocial interviews, immunological evaluations, and cervical swabs for HPV detection and subtyping were conducted on 36 HIV+ African-American, Haitian, and Caribbean women. RESULTS: Greater pessimism was related to lower NKCC and cytotoxic/suppressor cells after controlling for presence/absence of HPV Types 16 or 18, behavioral/lifestyle factors, and subjective impact of negative life events. CONCLUSIONS: A pessimistic attitude may be associated with immune decrements, and possibly poorer control over HPV infection and increased risk for future promotion of cervical dysplasia to invasive cervical cancer in HIV+ minority women co-infected with HPV. PMID- 9847031 TI - Antidepressants for the virtually eviscerated patient: options instead of oral dosing. AB - OBJECTIVE: Alternate ways of administering antidepressants when oral intake is impossible are discussed. METHOD: Reviews of 1) the medical circumstances that preclude oral medication administration and 2) novel administration strategies for antidepressants were conducted. RESULTS: Consultation psychiatrists not infrequently encounter depressed patients who lack a functioning gastrointestinal tract and who thus cannot absorb oral antidepressant medication. Under these circumstances, antidepressants can be administered intravenously, by rectal suppository, or topically. CONCLUSION: There are options for administration of antidepressant medication when oral intake is impossible. PMID- 9847032 TI - An investigation of the co-occurrence of panic and somatization disorders through temperamental variables. AB - OBJECTIVE: The relation of temperamental dimensions to somatization and panic disorders in women was investigated. METHOD: Eighteen patients with both panic and somatization disorders, 41 patients with panic disorder only, and 22 control subjects were compared on the Tridimensional Personality Questionnaire. RESULTS: The patients with somatization and panic disorders had significantly higher Novelty Seeking values than both the patients with panic only and the control subjects. The only significant correlation was between the number of symptoms of somatization disorder and Novelty Seeking. The presence of other psychiatric conditions, comorbid with panic disorder in the patient group, did not affect significantly the temperamental variables. CONCLUSIONS: A temperamental disposition characterized by extraversion, impulsivity, as well as frequent exploratory activity and pursuit of novel and pleasurable activities may help differentiate women with both somatization and panic disorders from women with panic disorder only. PMID- 9847033 TI - Anger expression and incident hypertension. AB - OBJECTIVE: It has long been thought that anger is important in the development of essential hypertension. However, tests of this hypothesis have yielded conflicting findings. This study prospectively examined the relationship between anger expression style and incident hypertension in a population sample of middle aged men. METHODS: Participants were 537 initially normotensive men from eastern Finland, who completed a medical examination and series of psychological questionnaires at baseline and at 4-year follow-up. Anger expression was assessed by Spielberger's Anger-out and Anger-in scales. RESULTS: At follow-up, 104 men (19.4%) were hypertensive (blood pressure > or = 165 mm Hg systolic and/or 95 mm Hg diastolic). Age-adjusted logistic regression analyses revealed that each 1 point increase in Anger-out was associated with a 12% increase in risk of hypertension after 4 years of follow-up (p < .002), which corresponded to a two fold increased risk of hypertension among men with scores in the top tertile of the Anger-out scale, relative to those with scores in the bottom tertile (odds ratio = 2.00, 95% confidence interval 1.20-3.38). Each 1-point increase on the Anger-in scale also was related to a 12% increased risk of hypertension (p < .01). Adjustments for body mass index, smoking, alcohol consumption, physical activity, a positive parental history of hypertension, and baseline resting diastolic blood pressure had little impact on the findings. CONCLUSIONS: These data provide strong epidemiological evidence for a positive relationship between anger expression style and subsequent hypertension, independent of known risk factors. Findings support the hypothesis that extreme expression of anger in either direction has adverse cardiovascular consequences. PMID- 9847034 TI - Blood pressure-related hypoalgesia in bulimia nervosa. AB - OBJECTIVE: This study examined pain sensitivity and its relationship to arterial blood pressure in bulimia nervosa (BN). METHODS: Fourteen women who met Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, criteria for BN, purging subtype, and 14 controls were tested for ischemic pain sensitivity after an extended baseline period. Blood pressure, cardiac output, stroke volume, and total peripheral resistance were assessed during baseline, during ischemic pain testing, and at the point of voluntary tolerance. RESULTS: Women with BN had significantly greater ischemic pain tolerance than controls. Additionally, only for BN women was blood pressure related to pain sensitivity. Systolic blood pressure during the pain procedure and at the point of tolerance was positively related to pain threshold and tolerance times and negatively related to rated unpleasantness of pain in BN, whereas no relationships involving blood pressure and pain sensitivity were observed in controls. CONCLUSIONS: These results may have implications for maladaptive changes in central pain-cardiovascular regulatory systems for women with BN. PMID- 9847035 TI - Chronic fatigue syndrome in psychiatric patients: lifetime and premorbid personal history of physical health. AB - OBJECTIVE: This preliminary report compares a group of chronic fatigue syndrome (CFS) patients and controls on several variables of potential significance in the etiology of CFS. METHOD: The lifetime prevalence of reported physical disorders was compared among 46 CFS psychiatric patients, 92 relatively physically healthy psychiatric patients (C-I), and 46 psychiatric patients selected without regard to physical health (C-II). All patients were matched on age, sex, and psychiatric diagnosis and were drawn from the same psychiatric practice. The same groups were compared on a 7-point scale of lifetime physical health by three raters independently evaluating physical health narratives of the CFS patients up to the time of onset of CFS and that of the controls up to the corresponding age. RESULTS: The CFS patients had a significantly higher reported lifetime prevalence of irritable bowel syndrome (IBS), infectious mononucleosis-like syndromes (IM), infectious mononucleosis-like syndromes two or more times (IM x 2), and herpes (other than genital or perioral herpes) than one or both control groups. The CFS group also had a higher incidence of allergic rhinitis or asthma, IBS, IM, and IM x 2 than the combined controls. On the independent ratings, the CFS patients had significantly more impaired physical health up to the time of onset of the CFS than C-I at a comparable age. CONCLUSIONS: The findings suggest that a general health factor may be involved in the pathogenesis of some cases of CFS. PMID- 9847036 TI - Depressive symptomatology and vital exhaustion are differentially related to behavioral risk factors for coronary artery disease. AB - OBJECTIVE: This study proposes to assess the differences of two psychosocial risk indicators for coronary artery disease (CAD), ie, depressive symptoms and vital exhaustion. METHOD: In a representative, stratified, nation-wide sample of the population of Hungary over the age of 16 years (N = 12,640), analyses were made of whether those risk indicators were differentially related to several illness behaviors (including history of cardiovascular treatment and cardiovascular sick days), cognitions, mood states, and socioeconomic characteristics that may generally be associated with increased CAD risk. The sample was stratified by age, sex, and composition of the population of all counties in Hungary. RESULTS: Although depressive symptoms and vital exhaustion correlated strongly, there were clear and significant differences in strength of association between depressive symptoms, vital exhaustion and several variables under study. Dysfunctional cognitions, hostility, lack of purpose in life, low perceived self-efficacy, illegal drug use, alcohol and drug abuse, several forms of subjective disability complaints and history of treatment because of congenital disorders, and chronic skin and hematological disorders were more often associated with depressive symptoms, whereas loss of energy, use of stimulants, chest-pain-related disabilities, history of treatment because of cardiovascular disorders, and self reported cardiovascular sick days were significantly more often associated with vital exhaustion. CONCLUSIONS: Vital exhaustion and depressive symptomatology are differentially associated with relevant external criteria. Vital exhaustion is associated with perceived cardiovascular complaints and history of cardiovascular treatment, whereas depressive symptomatology seems to be more closely connected to disabilities and complaints related to alcohol, drug, and congenital-disorder, and to dysfunctional cognitions and hostility. PMID- 9847037 TI - Fatigue in HIV illness: relationship to depression, physical limitations, and disability. AB - OBJECTIVE: This study was conducted to investigate the prevalence of clinical fatigue reported by gay/bisexual men at all HIV illness stages, and whether fatigue, while associated with depression, independently contributes to limitations in physical function and disability. METHOD: HIV- men, HIV+ men with CD4 counts >500, HIV+ men with CD4 counts 200 to 500, and men with AIDS were compared on prevalence of clinical fatigue, as defined by a standardized instrument. Among HIV+ men, the relationships among fatigue, depressed mood, major depressive disorder, HIV illness markers (including CD4 count and HIV RNA viral load), physical limitations, and disability were assessed at baseline and after 1 year. RESULTS: The prevalence of clinical fatigue in men with CD4 counts <500 was 14%, significantly higher than HIV- men and HIV+ men with CD4 counts >500. However, fatigue was not directly correlated with CD4 count or HIV RNA. Fatigue was a chronic symptom that was associated with depressed mood, major depressive disorder, physical limitations, and disability. After 1 year, an increase in depressive symptoms predicted a small amount of variance in fatigue; however, depressive symptoms were not associated with physical limitations or disability after controlling for fatigue. CONCLUSION: Fatigue is a chronic symptom that is more prevalent in advanced HIV illness, and which, although associated with depression, does not seem to be merely a symptom of depression. Because fatigue contributes independently to physical limitations and disability, it should be assessed and treated. PMID- 9847038 TI - Childhood attachment and loss experiences affect adult cardiovascular and cortisol function. AB - OBJECTIVE: This study proposes to test the hypothesis that early loss of a parent, coupled with poor quality family relationships, would result in long-term increased cardiovascular and cortisol reactivity to stress. METHODS: Subjects included 30 university students who lost one parent before age 16, and 31 control subjects. Blood pressure (BP) was measured continuously during 5-minute baseline and recovery periods, and during each of 2 tasks--viewing a 7-minute video clip depicting the death of a parent, and giving a 3-minute impromptu speech (1-hour rest between tasks). Salivary cortisol samples were collected immediately before each task, and at 5 and 20 minutes posttask. Quality of family relationships (FR) was measured using the Moos Family Environment Scale. RESULTS: Repeated-measures analysis of covariance revealed significant main effects on BP of both parental loss and FR for both tasks (all p values < .05) such that subjects who lost a parent or reported poor quality FR showed higher BP across all periods. The loss by FR by period interaction was not significant. An FR by period interaction was found for cortisol during the movie, in which poor quality FR subjects showed increased cortisol, whereas all others showed decreases. A loss by period interaction was found for cortisol during the speech, in which cortisol increased in loss subjects and decreased in non-loss subjects. CONCLUSIONS: These findings indicate that both childhood loss of a parent and poor quality of caretaking are associated with long-term increases in BP and altered neurohormonal responses to stress. PMID- 9847040 TI - Stimulus avoidance and perceptual adaptation: a vicious cycle paradigm? AB - Restricting the encountered range of stimuli on a particular stimulus dimension changes a person's perceptual frame of reference, or adaptation level, for such stimuli. The present experiment explores the effect of stimulus avoidance on perceptual adaptation and subsequent avoidance. Results converge with previous findings to indicate that avoidance of danger signals reduces perceptual adaptation level for those signals. In the present experiment, subjects first avoided high-pitched danger-signal tones, thus restricting the range of tone stimuli they encountered, and subsequently overestimated the pitches of low pitched safety-signal tones. They consequently avoided the low tones as if they were high-pitched danger signals. Perceptual adaptation was evident in subjects' tone pitch ratings only if they had rated the tones throughout their avoidance trials. PMID- 9847039 TI - Fitness and gender-related differences in heart period variability. AB - OBJECTIVE: The purpose of this study was to elucidate the processes of autonomic nervous system control of the heart in high fit and low fit men and women by examining indices of vagally mediated cardiac control. METHOD: There were 40 participants (19 women and 21 men) classified as either high fit or low fit on the basis of scores from the University of Houston Non-Exercise Test (N-EX). Both time domain and frequency domain cardiovascular measures were calculated during a resting baseline period (BL), a facial cooling task (FCT) designed to elicit parasympathetic activity, a reaction time task (RT) designed to elicit primarily sympathetic activity, and a combination task (COMBO) designed to elicit a combination of both parasympathetic and sympathetic activation and recovery periods after each task. RESULTS: Both time and frequency domain indices of heart period variability (HPV) indicated that high fit individuals exhibited greater vagal control of the heart relative to low fit individuals at baseline and across all tasks. In addition, the results suggest greater relative vagal control of the heart in women compared with men. Reactivity scores revealed significant main effects for condition for both time and frequency domain measures of HPV, but no main effects or interactions involving fitness or gender. CONCLUSIONS: First, small differences in aerobic capacity result in meaningful differences in cardiac autonomic control, with high fit individuals clearly demonstrating greater vagally mediated cardiac control. Second, important gender-related differences in HPV indicate that women exhibit greater vagal control and men exhibit more sympathetic cardiac control even though heart period (HP) did not differ. Third, the mechanisms underlying the beneficial effects of fitness and those placing women at a reduced risk for coronary heart disease (CHD) seem to be similar. PMID- 9847041 TI - Biological and psychological processes in recovery from depression during cognitive therapy. AB - This study investigated the relationship between biological and psychological processes in the recovery phase of depression during treatment with cognitive therapy. In particular, we tested the hypothesis that biological and psychological processes are independent; the hypothesis that they are related in a linear or sequential process; and the hypothesis that there is a circular relationship between them. 35 depressed patients completed a 12 week program of cognitive therapy. Changes in measures of negative thinking and amine dysfunction over the course of therapy were compared for those who improved and those who did not. There was a significant relationship between improvement in symptoms and change for ATQ scores, DAS scores, and epinephrine levels. There was significant change in metanephrine levels during therapy which was not related to improvement. The changes were not consistent across indices. Although the results did not provide definite support for any of the four models of the relationship between biological and psychological variables during recovery from depression, the finding that there was some change in some of the indices of amine dysfunction during cognitive therapy indicates that the interface between biological and psychological processes in depression should continue to be studied. PMID- 9847042 TI - Characteristics of early unpleasant dental experiences. AB - There is evidence in the literature that dentally anxious individuals are not a homogeneous group. Various ways of categorizing them have been suggested. These categories have stemmed primarily from clinical experience or psychiatric diagnoses. In addition, Reiss' (1987, 1991) expectancy theory of fear, anxiety and panic appears to fit some of the etiological data. At the simplest level, it is suggested that dental anxiety may result from direct conditioning experiences or from a constitutional vulnerability to develop anxiety-based problems. Taking conditioning theory and Reiss's model as a basis, it was predicted dental events deemed unpleasant would fall under four categories: (a) events relating to dental specific procedures and stimuli, (b) events relating to injuries, (c) events relating to emotional responses and (d) events relating to dental personnel behavior. In addition, it was predicted that individuals who would report high anxiety sensitivity as measured by the anxiety sensitivity index (ASI) would be more likely to perceive unpleasant incidents relating to injuries, emotional responses and dental personnel behavior than individuals with low-anxiety sensitivity. On the other hand, everyone would be affected by incidents relating to specific dental procedures and stimuli. The participants were 510 university students who were surveyed during class time. The first prediction was supported but the second one was only partially supported, in that, participants who obtained high scores on the ASI differed from the others only regarding injuries. PMID- 9847043 TI - Devices and methods for administering carbon dioxide-enriched air in experimental and clinical settings. AB - Although researchers successfully have used carbon dioxide-enriched air in experimental and clinical preparations, its functional properties may differ across laboratories due to procedural differences. Additionally, current procedures may be too simplistic for more complex experimental designs. To address these issues, we present three devices for administering carbon dioxide enriched air. Although these devices differ concerning variables such as mode of operation, ease and cost of implementation, and complexity of experimental designs that may be undertaken, a reasonable level of standardization may be achieved because the inhalations experienced by participants are functionally equivalent across devices. We discuss advantages and disadvantages of these devices regarding experimental panic provocation and aversive conditioning preparations. PMID- 9847044 TI - Case report: treatment of a woman with alcohol and binge eating problems. AB - The prevalence of eating disorders generally, and bulimia symptomatology in particular, in problem female drinkers has been consistently reported. According to an addictive behaviour model, these two disorders share common features, in particular a compulsion to engage in the behaviour and a sense of loss of control. However, there has been no attempt to treat sufferers of both conditions with a similar treatment approach. The present case study illustrates how a cognitive behavioural and exposure-based approach successfully treated a woman with a history of bulimia and alcohol problems. At 12 months follow up, there was a significant reduction in alcohol consumption and an absence of bingeing and purging episodes. PMID- 9847045 TI - Habit-reversal treatment for children's stuttering: assessment in three settings. AB - Two boys (9 and 14 years old) participated in an evaluation of treatment for stuttering. Habit reversal procedures (awareness training, regulated breathing, and social support) were combined with teaching positive attitudes to parents. During treatment, stuttering frequencies decreased, speech rates increased, and speech naturalness ratings increased. These changes occurred in the clinic, the child's home, and the child's school. After treatment, stuttering remained low in the clinic and at home, but increased stuttering was found at school. PMID- 9847046 TI - Common treatment goals of antipsychotics: acute treatment. AB - When a patient with an acute exacerbation of schizophrenia is admitted into the hospital, the target symptoms include pathologic excitement/agitation and exacerbated psychotic symptoms. The goal of hospitalization becomes attenuation of these symptoms to a level compatible with safe discharge. The mainstay of stabilization is antipsychotic treatment. A risk/benefit analysis of the conventional versus the newer antipsychotics favors the use of the newer agents as first-line drugs. These newer antipsychotic agents represent the first significant advance in the pharmacologic treatment of schizophrenia in the past four decades. They are at least as effective as conventional agents and are clearly superior from a safety perspective. Because of short inpatient stays, the challenge for clinicians is to provide an adequate treatment period without aggressively escalating the dose. PMID- 9847047 TI - Pharmacologic treatment of first-episode schizophrenia: early intervention is key to outcome. AB - The early recognition and management of a first episode of schizophrenic illness is a difficult task, with identification complicated by a broad differential diagnosis, lack of definitive data on the prognostic implications of premorbid/prodromal symptoms, and, until recently, treatment limited to pharmacologic agents with severe adverse effects. The first psychotic episode in patients with schizophrenia is the most responsive to treatment in terms of both rate and degree. However, first-episode patients are also more likely to develop motor side effects, even at lower medication doses, than multiepisode patients. Considerable evidence supports the assertion that early treatment can improve outcome and possibly prevent the development of full-blown illness in high-risk individuals. There is evidence that atypical antipsychotic medications are effective in the treatment of first-episode schizophrenia and are well tolerated. The improved tolerability associated with the newer antipsychotic medications, including a lower risk for motor side effects and possible lower risk for development of tardive dyskinesia, has swung the risk-benefit balance in favor of early and aggressive treatment. By intervening early and providing long-term maintenance treatment, the course of schizophrenic illness may be altered in the coming years with overall decreased deterioration and chronicity and overall improved functioning resulting in lower societal costs. PMID- 9847048 TI - The routine use of atypical antipsychotic agents: maintenance treatment. AB - Placebo-controlled maintenance studies of conventional antipsychotic agents demonstrate a significant reduction in the risk of schizophrenic relapse in neuroleptic-treated patients. Neuroleptic discontinuation even in patients who remained in remission for as long as 5 years results in a relapse rate comparable to that seen for patients initially assigned to placebo. Yet, patients maintained on conventional neuroleptics are exposed to the risk of tardive dyskinesia (approximately 5% per year for patients with up to 10 years of neuroleptic exposure). Attempts have been made to reduce neuroleptic exposure. A lower maintenance dose was associated with higher relapse rates, as was intermittent, targeted therapy. Psychoeducational treatment studies reaffirmed that the major influence on the rate of rehospitalization was the dose of conventional maintenance medication. Although data are scarce for maintenance treatment with atypical antipsychotic drugs, findings suggest that atypical agents are at least as efficacious and may be better tolerated. Olanzapine has demonstrated efficacy in maintenance treatment as well as a reduced risk of tardive dyskinesia compared with haloperidol. PMID- 9847049 TI - Formulary decisions and health economics. AB - Because of increasing concerns about health care costs, physicians must consider the cost-effectiveness of a treatment strategy, as well as its efficacy and safety. The question of whether the greater expense of a newer drug is justified over the cost of a generic drug deserves a comprehensive evaluation. The determination of effectiveness and tolerability of the newer antipsychotics should be expanded to include quality-of-life issues, reintegration of the patient into the community, resource utilization, and medical costs. There are clear indications that patients who take atypical antipsychotics utilize fewer medical resources than patients who take typical antipsychotics; however, the positive outcomes of the newer drugs must be translated into cost benefits if formularies are to be intelligently controlled. PMID- 9847050 TI - Management of chronic psychotic ambulatory outpatients. AB - The introduction of novel atypical antipsychotic medicines has raised new possibilities in the treatment of psychotic patients. In particular, the discovery of pharmacologic agents that may ameliorate the most stubborn positive and also negative symptoms without adding the burden of major side effects is revolutionizing treatment expectations. However, it is also becoming abundantly clear that successful treatment requires more than just the administration of a medicine. Treatment must also address the inner experiences of the patient, as well as the social and psychological handicaps that are associated with the illness. Some of the relatively neglected complications of using typical antipsychotic medicines include hyperprolactinemia and antipsychotics that may require concomitant treatment with anticholinergic agents, which themselves have an array of side effects. This article presents a detailed method for deciding when and how to use the new medications effectively and how to manage the transition from one medicine to another. PMID- 9847051 TI - Long-term considerations after switching antipsychotics. AB - The so-called "atypical" antipsychotics are rapidly becoming the de facto standard pharmacologic treatment of schizophrenia. This article reviews some common psychopharmacologic and psychological issues that may arise after an outpatient with schizophrenia is switched to one of the newer antipsychotics. Important issues to consider in the first few months after switching include assessment of response to the new medication, dealing with subsequent psychological reactions, and management of an unsatisfactory response. Once the response is established, there are other pharmacologic and psychological issues that arise during the next year or two. Pharmacologic issues that emerge later on include the role of long-term combination antipsychotics, management of new side effects, and deciding whether and when to switch again. Some of the long-term psychological issues include changes in self-image that arise from being less visibly ill, sexuality and intimacy concerns, and recovery issues. PMID- 9847053 TI - Reverse physiology: discovery of the novel neuropeptide, orphanin FQ/nociceptin. AB - The search for novel neurotransmitters and neuropeptides has been recently revolutionized by the development of a purification strategy based on orphan G protein-coupled receptors, cloned receptors for which no natural ligands are known. This strategy uses the orphan receptor as bait to identify its natural ligand. This article will review the discovery of the first natural ligand isolated following this strategy. This ligand is a peptide that shares some striking sequence similarity to the opioid peptides and has been named Orphanin FQ or Nociceptin (OFQ/NOC). The discovery of OFQ/NOC will be described as one example of the use of orphan receptors in identifying novel neurotransmitters and neuropeptides, an example that has already been followed in the identification of other novel neuropeptides. After reviewing the conceptual and technological basis of the strategy and its successful first application, we discuss the criteria used to validate OFQ/NOC as the natural ligand of the orphan receptor and as a genuine neuropeptide. We also discuss the importance and implications of discovering OFQ/NOC mode of synthesis, which is synthesized as expected in the form of a larger polypeptide precursor, which in turn raises the question of the existence of other OFQ/NOC-related peptides. We then present an overview of the numerous studies that have blossomed after the OFQ/NOC discovery and describe the numerous physiological roles that have already been attributed to OFQ/NOC, and in particular the controversy regarding its involvement in pain perception. Because of the similarities between the OFQ/NOC and opioid systems, we also discuss overlaps between these systems and present evidence favoring a pharmacological separation between these systems. We finish by outlining the power of the orphan receptor strategy and by discussing some of its pitfalls. PMID- 9847052 TI - Management of the agitated elderly patient in the nursing home: the role of the atypical antipsychotics. AB - Agitation is a frequent and troublesome problem in the long-term care setting. The term agitation is a nonspecific descriptor of a variety of verbal, vocal, and motor behaviors that can be unsafe, disruptive, and distressing to staff, families, and patients alike. Agitation can occur as a result of psychiatric and nonpsychiatric conditions, and appropriate treatment needs to be directed at the target symptoms. Optimal results are achieved with a combination of behavioral and pharmacologic interventions. In this review, we examine some of the causes and interventions that can assist physicians caring for the agitated elderly in long-term care settings. The role of the atypical antipsychotics is discussed in detail. PMID- 9847054 TI - Developmental aspects of NMDA receptor function. AB - Glutamate is the major excitatory neurotransmitter in mammalian synapses. It binds to three classes of predominantly postsynaptic ionotropic receptors to activate receptor-associated channels, and a class of metabotropic receptors to activate G-protein mediated transduction pathways. The N-methyl-D-aspartate (NMDA) receptor (NR) is distinctive in being both ligand and voltage-gated, and selectively permeable to Ca2+. As a consequence, NR-mediated alterations in intracellular Ca2+ levels regulate a variety of signaling pathways, ranging from localized, acute effects on receptor and channel activities to long-term effects on nuclear gene transcription. Regulated expression, assembly, and modulation of distinct heteromeric NR complexes comprised of different subunit combinations contributes to this functional diversity. NRs have been implicated in several developmental processes, and evidence supporting their role in migration, survival, and synaptic maturation is discussed. PMID- 9847055 TI - Felbamate: clinical and molecular aspects of a unique antiepileptic drug. AB - Felbamate was launched in 1993 in the U.S. as a "new generation" antiepileptic drug (AED) with a unique mechanism of action. It proved efficacious in patients refractory to other AEDs and was particularly beneficial in children suffering from Lennox-Gastaut syndrome, being the first drug shown to be effective at treating this condition in controlled trials. Following the occurrence of rare cases of aplastic anemia and of hepatic failure associated with the use of felbamate during early 1994, a "black-box" warning was added to the drug's package insert. Despite this, felbamate continues to be used in many patients, although not as a first-line treatment. Felbamate's dual mechanism of action- enhancing the GABA system while inhibiting excitatory amino acid responses--may explain its efficacy in a broad range of epileptic patients. A better understanding of this mechanism may lead to the development of felbamate-like drugs with a better side effect profile. PMID- 9847056 TI - Nitric oxide in cerebral ischemic neurodegeneration and excitotoxicity. AB - The observation that the free radical nitric oxide (NO) acts as a cell signaling molecule in key physiological processes such as regulation of blood pressure and immunological host-defense responses is probably one of the most important and exciting findings made in biology in the last decade. Likewise, in the brain NO has been implicated in a number of fundamental processes, including memory formation, sexual behavior and the control of cerebral blood flow. This has radically altered the accepted dogma of brain physiology and has placed NO at the center stage of neuroscience research. Evidence suggests that some of the actions of NO in the brain may be intimately linked to those of the classic excitatory neurotransmitter glutamate. The historical view that aberrations in glutamate signal transduction may underlie central neurodegeneration following, for example, cerebral ischemia, has implicated NO, by default, as a potential mediator of neuronal death. Indeed, with the advent of potent and specific compounds that interact with NO synthesizing (NOS) enzymes and with the NO signaling cascade, there is now ample evidence to suggest that NO can mediate neurodegeneration, although its involvement is paradoxical. Its cerebrovascular effects may act to limit ischemic damage by preserving tissue perfusion and preventing platelet aggregation, while NO produced in the parenchyma, either directly following the ischemic insult or at a later stage as part of a neuroinflammatory response, may be deleterious to the outcome of ischemia. Nonetheless, significant efforts are made into the potential therapeutic use of chemical NO donors and specific NOS inhibitors in the treatment of cerebral ischemia and other central neurodegenerative disorders. Here, the latest concepts and developments in our understanding of the role of NO in cerebral ischemic neurodegeneration are discussed. PMID- 9847057 TI - Is the caudomedial shell of the nucleus accumbens part of the extended amygdala? A consideration of connections. AB - The conceptual origins of two forebrain functional-anatomical systems, the ventral striatopallidum and extended amygdala, are reviewed briefly, and reductionist and constructionist doctrines pertaining to descriptions of forebrain organization are generally discussed briefly. The cortical, subcortical, and intrinsic connectional relationships of the caudomedial shell of the nucleus accumbens, an anomalous component of the ventral striatopallidum, are compared with those of the extended amygdala, leading to an attempt to formulate an assessment as to the extent to which the caudomedial shell might actually be regarded as a part of extended amygdala, or a transitional structure with a capacity to bridge extended amygdaloid and striatal mechanisms. The relatively distinct intrinsic circuitries and different outputs of the two are noted. Extended amygdala projects most strongly to the posterior half of the lateral hypothalamus and brainstem autonomic effector sites. Caudomedial shell and ventromedial ventral pallidum project throughout the lateral preoptico-lateral hypothalamic continuum and are major components in a basal ganglia thalamocortical pathway to the prefrontal cortex. It is concluded that connectional features distinguishing the caudomedial shell/ventromedial ventral pallidum and extended amygdala are of sufficient magnitude that the two should most usefully be regarded as separate, interacting functional-anatomical entities. PMID- 9847058 TI - Evolving technique of percutaneous nephrolithotomy in a developing country: Singapore General Hospital experience. AB - At our institution, percutaneous nephrolithotomy (PCNL) is the preferred treatment modality for renal calculi with a large stone burden (example, staghorn calculi) and renal calculi which have failed extracorporeal shockwave lithotripsy (SWL). The aim of this paper is to present the technical refinements in PCNL and the results obtained by a single surgeon. From June 1995 to July 1997, 135 PCNL procedures were performed on 114 renal units. The mean age of the patients in the series was 51 years. The PCNL was performed on 28 complete staghorn calculi (25%), 24 partial staghorn calculi (21%), 25 large renal calculi (22%), 26 impacted ureteropelvic junction calculi (22%), 5 impacted ureteral stones (4%), and 6 symptomatic lower-pole calculi (5%). Most patients (97%) had a successful outcome (82% stone-free and 15% only insignificant residual fragments). Of the four failures, three had previous open renal split operations. The average postoperative stay was 5.2 days. Only one patient (0.7%) had urosepsis, and another had an arteriovenous fistula and was the only patient to require blood transfusion in our series. Only 21% (n = 24) required SWL after PCNL, and one patient required ureteroscopy. Technical refinements in PCNL include: (1) single stage PCNL with percutaneous access obtained by the urologist in the operating room; (2) flexible endoscopy to inspect the whole pelvicaliceal system systematically; (3) liberal use of secondary PCNL to ensure stone-free status; and decreased reliance on SWL after PCNL. The trend is to rely on PCNL monotherapy to manage staghorn calculi rather than PCNL and SWL combination therapy. PMID- 9847059 TI - The supracostal percutaneous nephrostomy for treatment of staghorn and complex kidney stones. AB - From March 1995 to May 1997, 104 patients underwent 115 supracostal percutaneous nephrolithotomy (PCNL) procedures for the treatment of 102 complete staghorn calculi, 6 large semistaghorn calculi, 3 large upper-caliceal stones, and 4 significant volumes of residual stone fragments after SWL. Additional renal access was required mainly for complete staghorn stones (23 patients; 20%). Extracor poreal lithotripsy was performed in 30.4% of cases, and second-look PCNL was done in 15.6%. The stone-free rate was 87%, and the infection-free rate at 7 to 33 months was 88.5%. Among 115 supracostal PCNL procedures, complications were encountered in 10 (8.7%). These problems included four large pleural effusions that were drained by chest tube in three patients and by repeated thoracocentesis in one patient. Six patients developed significant atelectasis, which was treated by vigorous physiotherapy in five and flexible bronchoscopy in one. We conclude that the supracostal approach provides direct and optimal access to most staghorn calculi with an excellent stone-free rate. The advantages of this approach can be achieved with a slight and acceptable increase in morbidity. PMID- 9847060 TI - New visualization techniques for in utero surgery: amnioscopy with a three dimensional head-mounted display and a computer-controlled endoscope. AB - Endoscopic fetal surgery may reduce preterm labor associated with open hysterotomy but is partially limited by current visualization technology. We investigated a three-dimensional (3D) imaging system coupled to a head-mounted display (3D-HMD) and also employed a computer-controlled zoom endoscope for noninsufflated amnioscopy. Pregnant sheep were prepared in aseptic fashion for general anesthesia. Uterine access was obtained following maternal laparoscopy. A 10-mm zoom endoscope (Vista Medical Technologies, Carlsbad, CA) was used to examine the fetus and uterine contents. Fetal limbs were exteriorized for microsurgery. A new system (Vista Medical Technologies) was attached to an operative microscope, permitting projection of a 3D image via an HMD. The fetus and umbilical cord were inspected using the zoom endoscope, which changes the depth of focus under computer control. Basic manipulations of the fetus and cord were easily completed. Real-time 3D fetal imaging was accomplished. The added depth perception enabled detailed fetal and placental examination, fostering manipulation of the fetus and cord. The HMD was adjusted to fit several surgeons, permitting a natural operative posture. This unit has the capacity to display any video, CT, MR, or ultrasound image as a picture-in-picture. The success of minimally invasive fetal surgery is in part dependent on the development of video technologies capable of providing both magnification and optimal resolution. The zoom endoscope affords excellent visibility of multiple surgical targets without instrument repositioning. A 3D HMD system such as this provides greater anatomic detail and an appreciation of fetal movements that may make intrauterine procedures more feasible. PMID- 9847061 TI - Three-dimensional endoluminal ultrasonography of the ureter. AB - Endoluminal ultrasonography has been useful in demonstrating the ureteral and periureteral anatomy. Devices now available give cross-sectional images. A computer program is available to reconstruct these images into three-dimensional (3D) representations. Endoureteral sonography was performed using 6.2F endoluminal ultrasound catheters connected to a dedicated ultrasound unit giving cross-sectional images. The images along the segment in the study were collected by advancing the catheter to the most proximal portion and pulling it in antegrade fashion through the segment with a step-pulling device. The images were collected on videotape and reconstructed into 3D representations using a dedicated 3D computer system (Lis 6000-A). Anatomic features studied included ureteral strictures, obstructed ureteropelvic junction, ureteral neoplasms, and the ureterovesical junction. In each case, additional information and appreciation of the structure could be obtained with the 3D reconstruction. Endoluminal ultrasonography has provided an additional dimension to imaging of the ureter. Three-dimensional reconstruction of these images can give additional information regarding the longitudinal appearance of ureteral segments without the need for development of new ultrasound transducers. It can offer appreciation of anatomic features not evident with two-dimensional imaging alone. PMID- 9847062 TI - Ex vivo comparison of four lithotripters commonly used in the ureter: what does it take to perforate? AB - We hoped to determine the number of pulses and energy needed to create acute ureteral perforations with four different lithotripters in a reproducible ex vivo model. A simple model was constructed to control variables in the testing such as wall thickness, intraluminal pressure, distance between the probe tip and ureter, and power delivered to tissue. Segments of domestic pig ureter were prepared and fixed in position in a normal saline (NS) bath at room temperature. We then attempted perforation with the holmium:YAG (HoL) laser, coumarin pulsed-dye laser (CdL), electrohydraulic lithotripter (EHL), and pneumatic impactor (PI) by placing the instrument probes at right angles to the ureteral wall. The ureter was filled with a methylene blue-stained solution of NS at 90 cm H2O pressure via a urodynamics catheter, and perforation was recorded on initial extravasation of dye. The endpoints measured were time to perforation and total energy required. At 0.5 mm of separation between the wall and probe, the HoL perforated the ureter in an average of 2 seconds and 0.01 kJ delivered at 5 W (10 Hz and 0.5 J/pulse). The EHL perforated at an average of 24.44 +/- 8.77 seconds and a total energy of 0.01 +/- 0 kJ. The CdL was able to perforate but at much longer intervals (257.51 +/- 99.08 seconds) and higher energy levels (12.88 +/- 4.95 kJ) on average than either the EHL or HoL. Lastly, the PI was unable to perforate the ureter in more than 6 continuous minutes of application. In addition, we found that at 2-mm separation between the HoL probe tip and the ureteral wall, acute perforation was not possible even at very high power settings. We conclude that although each endoscopic lithotripter has advantages as well as disadvantages, in this ex vivo model, it was clear that the HoL and EHL can easily perforate the ureter and must be used with vigilance. It was found that at 2 mm of separation between the probe and target, the HoL, was unable to perforate acutely. The CdL and PI were associated with a much higher safety index, and the PI was unable to produce ureteral perforation. PMID- 9847063 TI - Endobrst: simple and safe is best. AB - Minimal-access treatment for ureteropelvic junction (UPJ) obstruction is becoming increasingly complex. Is there still a place for the simple procedure of balloon dilatation? To examine the symptomatic and renographic results of patients at least 6 months after balloon disruption of an obstructed UPJ, we carried out a prospective audit of outcome as judged by DTPA renograms and clinical follow-up. The procedure has been performed on a total of 21 renal units (20 patients) with follow-up of 6 to 30 months (mean 22 months). Of these, 18 (86%) were asymptomatic at 3 months, although this success rate dropped to 17 (81%) by 6 months. Renographic split function improved in 11 renal units, and excretion improve in 14. Perioperative complications were few and associated with stent insertion. Three of the patients in whom the procedure failed went on to nephrectomy (14%); all had poor renal function (split <20%) at presentation. Although the quoted success rates are lower for endoluminal balloon rupture of stenosis treatment than open pyeloplasty, the former technique has significantly less morbidity. Our nephrectomy rate reflects our less than ideal early patient selection, and our balloon dilation technique is not recommended as a salvage procedure for patients in whom nephrectomy is inevitable. The learning curve is quick to climb, and this is a fundamentally simple procedure with medium-term results that remain encouraging. PMID- 9847064 TI - Ureteropelvic junction obstruction and ureteral strictures treated by simple high pressure balloon dilation. AB - The long-term results of simple high-pressure balloon dilation in the treatment of ureteropelvic junction obstruction (UPJO) and ureteral strictures were evaluated. A total of 77 consecutive patients were treated: 40 had UPJO and 37 ureteral strictures. The etiology of the obstruction included congenital UPJO, previous stones, sequelae of endoscopic and open surgery, radiotherapy, and urinary tract reconstruction. A retrograde ureteroscopic approach was used. Evaluation included clinical and radiographic examinations and renal scintigraphy with diuretic wash-out. The procedure was repeated in 21 cases. The median follow up was 29 months. The procedure was considered successful if it left the patient asymptomatic and with improved renographic function and drainage. The overall success rate was 70%. The best results were obtained in strictures secondary to stones, with a success rate of 94%, and in strictures secondary to reconstructive and ureteroscopic surgery, with a success rate of 91%. In congenital UPJO, the results were less encouraging: in patients with a symptom debut after the age of 18 years, balloon dilation was successful in 57% of cases; in patients with symptom debut before the age of 18 years, success was achieved in only 25% of cases. There were no major complications. It was concluded that simple high pressure balloon dilation is a safe and reasonably effective technique for the management of most ureteral strictures and congenital UPJO with symptom debut in adult life. Balloon dilation seems to have no place in the treatment of primary congenital UPJO in children. PMID- 9847065 TI - Complications of acucise endopyelotomy. AB - Endoscopic management of ureteropelvic junction (UPJ) obstruction has a success rate of 80% to 86%. We have been performing a ureteral cutting balloon procedure under fluoroscopic control (Acucise endopyelotomy) for UPJ obstruction at Loyola University Medical Center since 1991. The overall success rate in 77 patients was 78%. All patients had a preoperative intravenous urogram or a retrograde pyelogram, but none had vascular imaging studies. Acucise endopyelotomy consisted of a posterolateral incision of the UPJ and placement of an endopyelotomy or double-J stent. Foley catheter placement at the end of the procedure demonstrated significant gross hematuria in three patients (4%). All three remained hemodynamically stable but with significant drops in postprocedure hemoglobin levels, which necessitated blood transfusion. Aggressive management included angiographic studies and embolization of lower-pole branching arteries in two patients (3%). One patient stopped bleeding after being given two units of blood. None of the patients required an open exploratory procedure. Although the risk of vascular injury is low with Acucise endopyelotomy, prolonged postoperative gross hematuria does mandate investigation and observation. Angiographic embolization appears to be the therapeutic modality of choice for patients with hemorrhagic complications after an Acucise endopyelotomy. PMID- 9847066 TI - Transperitoneal laparoscopic nephrectomy in children. AB - Over a 40-month period, 24 consecutive children (1-15 years) underwent laparoscopic nephrectomy. The indication for surgery was a poorly functioning kidney (<6% on DMSA isotope scanning) secondary to a variety of causes, with or without pain or infection. Four cannulas were used in each patient. The kidney was approached through a small incision in the upper paracolic gutter without mobilization of the colon. The procedure was successful in all but one child, who had conversion to open technique because of poor laparoscopic viewing. In 12 children who required nephroureterectomy, the distal ureter was approached by an open technique through either a small extension of the iliac fossa cannula site or a Pfannenstiel incision for combined bladder surgery. The average operating time for laparoscopy was 85 (range 40-160) minutes. The children undergoing nephrectomy or nephroureterectomy alone had an average hospital stay of 2 (range 1-4) days. There were no laparoscopic or surgical complications. Laparoscopy provides a safe and successful approach to pediatric nephrectomy. The technique combines well with an open approach to the distal ureter when nephroureterectomy is indicated. PMID- 9847067 TI - Utility of the harmonic scalpel for laparoscopic partial nephrectomy. AB - Laparoscopic partial nephrectomy (LPN) remains a technically challenging procedure largely because of the lack of methods for obtaining consistent parenchymal hemostasis. The objective of this study was to determine if the extent of resection influences the ability of the harmonic scalpel to achieve hemostasis and to define the cases in which the harmonic scalpel is appropriate for LPN. Thirty LPNs were performed in a 25-kg domestic pig model. The blunt blade of the laparoscopic harmonic scalpel (LaparoSonic Coagulating Shears; Ethicon Endo-Surgery, Cincinnati, OH) at power level 5 was used to divide the parenchyma. Control of the renal hilar vessels was not obtained. Three standardized types of resections were performed: I = peripheral wedge biopsy; II = upper- or lower-pole nephrectomy; and III = heminephrectomy. Bleeding was graded on a scale from 0 to 4: 0 = no hemostasis; 1 = steady bleeding; 2 = moderate bleeding; 3 = parenchymal oozing; and 4 = dry. Hemostasis grades of 2 or less were clinically significant bleeding necessitating supplemental coagulation. The mean hemostasis scores showed a significant (P < 0.02) trend toward inadequate hemostasis with increasing extent of resection: 3.3 for Type I, 3.0 for Type II, and 2.4 for Type III. The percent of kidneys with grade 2 bleeding or worse was 9% for Type I surgery, 25% for Type II, and 57% for Type III. Successful hemostasis with the harmonic scalpel correlates with the extent of parenchymal resection in the porcine model. Most wedge excisions can be done with the harmonic scalpel alone, whereas larger resections necessitate supplemental coagulation. On the basis of this study, heminephrectomies with the harmonic scalpel are not recommended because of the high incidence of significant hemorrhage. PMID- 9847068 TI - Pediatric laparoscopy for nonpalpable testes with new miniaturized instruments. AB - New miniaturized pediatric telescopes (1.9 or 3.5 mm) and reusable instruments (3.5-mm trocars, scissors, graspers) were used for the first time in a prospective study to evaluate handling safety and efficacy in laparoscopic diagnosis of unilateral nonpalpable testes. The results were confirmed during the same anesthesia by open operation for either orchiopexy or removal of rudimentary spermatic cord structures. Laparoscopy in 13 boys revealed one abdominal testis, 5 vanishing testes, and 7 cases of inguinal retention associated with an open inner inguinal ring. The illumination and detail resolution of the minitelescope were excellent. The 3.5-mm instruments were fully functioning with regard to tissue dissection. No complications occurred. Without technical disadvantages but with increased safety for the patient, the new miniaturized pediatric laparocopic instruments indeed realize a step forward to minimal invasion in infants and children. PMID- 9847069 TI - In vitro results with special plastics for biodegradable endoureteral stents. AB - Internal ureteral stents are widely used in urologic practice for temporary urinary diversion, but all double-J catheters to date exhibit the same disadvantage; that is, they have to be removed endoscopically, necessitating further intervention. We tested different materials (designated G100X-15xLB and G100X-20xLB) to develop a biodegradable (biodissolvable) endoureteral stent that can be held in place without functional loss yet could be dissolved by changing the environment. The principle of the biochemical background is based on the physiological milieu of the urine. The plastics tested are stable in acidic and dissolve in alkaline conditions. In a first step, specimens of two polymers were placed in artificial urine of different pH over a period of 60 days and monitored for integrity (solution trial). In a second step, artificial urine was set in motion (744 mL/24 hours) an infusion pump (Volumed microVP 5000; Fresenius AG, Bad Homburg vdH, Germany) through an infusion set in which a 30-cm piece of the materials to test had been placed (ureter model). Below the inserted specimen, the lumen of the infusion tube was minimized to make obstruction by fragments more possible. In the solution trial, all specimens remained stable under physiologic conditions (pH 5.2) over a period of at least 30 days. The specimens dissolved completely when the pH was adjusted to an alkaline one (pH 7.9). In the ureter model, with pH values of 7.9, all specimens were decomposed after 20 hours, and no occlusion of the model occurred. Using acidic artificial urine, the specimens remained stable with a smooth consistent surface. The dissolution was not a standard chemical one; the materials broke into microscopically small pieces, with fragments of G100X-20xLB being smaller than those of G100X-15xLB. Our first in vitro results show that the tested materials are suitable for further development of biodissolvable endoureteral stents, dissolution of which can be steered by changing the urinary pH. PMID- 9847070 TI - Holmium laser enucleation of the prostate (HoLEP) combined with transurethral tissue morcellation: an update on the early clinical experience. AB - The preliminary experience with 64 patients who have undergone HoLEP combined with intravesical morcellation is presented. The mean preoperative prostate volume was 75.3 cc. The mean laser time was 46.9 minutes and the morcellator time a further 10.5 minutes. A mean total of 35.5 g of tissue was obtained, and 93% of the patients were discharged the day after their surgery without a catheter. The American Urological Association Symptom Score at 1 month was 8.6 and the peak flow rate 23.4 mL/sec. This combination of procedures allows prostate glands of virtually any size to be safely treated transurethrally, and a complete anatomic enucleation of the prostatic adenoma is achieved. PMID- 9847071 TI - Transurethral prostate ablation using saline-liquid electrode introduced via flexible cystoscope. AB - An interstitial saline-electrode has been developed to couple radiofrequency (RF) energy to prostate tissue in order to produce large lesions quickly and controllably. In this study, we attempted to produce similar results using a flexible catheter introduced transurethrally via a flexible cystoscope. An insulated 4F catheter with an extendable needle electrode (26-gauge, 5-mm exposed metal tip) was introduced through a port of a flexible cystoscope into the urethra of 10 dogs via a small perineal incision (required for the canine U shaped urethra). Under visual endoscopic guidance, the electrode was placed in the center of the prostate and the needle electrode extended at an angle into the prostate, one lobe at a time. Hypertonic saline (14.6% NaCl) was infused at 2 mL/min through the needle electrode into the tissue for 30 seconds before and during RF application. The energy (50 W, 475 kHz) was delivered for 30, 45, 60, or 90 seconds (RFT System; U.S. Surgical Corporation). Prostate and urethral temperatures were monitored using thermocouples mounted on the catheter. There was an automatic high-temperature shut-off if the urethra reached >42 degrees C or if impedance reached >300 omega during RF energy application. Prostate sizes ranged from 1.4 x 2.0 x 1.4 cm to 5.0 x 4.5 x 4.2 cm. In this group, there were no automatic shut-offs at any of the energy durations, and the lesions produced ranged from 1.3 x 1.3 x 1.1 cm (0.99 cm3) to 1.6 x 2.7 x 2.2 cm (5.03 cm3). The average ablation volumes produced were 1.76 cm3 at 30 seconds' RF application, 2.42 cm3 at 45 seconds, 3.96 cm3 at 60 seconds, and 5.03 cm3 at 90 seconds. Histologic examination revealed typical coagulation necrosis in the treatment zone without tissue desiccation. Introduction of the saline-electrode through a flexible cystoscope provides a minimally invasive procedure that quickly and controllably produces large lesions that may provide effective treatment for benign prostatic hyperplasia with less patient discomfort. PMID- 9847072 TI - Alterations in predicted growth rates of pediatric kidneys treated with extracorporeal shockwave lithotripsy. AB - The long-term effects of extracorporeal shockwave lithotripsy (SWL) on the kidneys of children treated for renal calculi are unclear. In order to determine if SWL has any negative effects on renal growth rates, we reviewed long-term (mean 9-year) follow-up data on 29 pediatric patients treated between 1984 and 1988 with an unmodified Dornier HM3 lithotripter. Changes in renal length, serum creatinine, and blood pressure were analyzed. Predicted renal growth was calculated using a formula for age-adjusted renal length. Treated kidneys were stratified into normal and abnormal groups based on a history of renal surgery, evidence of recurrent infection, and obvious anatomic abnormalities. Fifty-six upper urinary tract calculi were treated in 34 renal units. Twenty-two renal units (68%) were rendered stone free, and 65% of the patients continue to be stone free. At follow-up, one patient was classified as having new-onset hypertension, and the mean serum creatinine was 0.93 +/- 0.08 mg/dL. Both at treatment and at follow-up, no significant differences were found in the sizes of the treated and untreated kidneys. However, at treatment, the abnormal group of kidneys seemed to be smaller than expected (mean Z -1.30 +/- 1.10), whereas the group of normal kidneys was very close (mean Z 0.18 +/- 0.54) to the predicted length. At follow-up, the deviations between actual and predicted renal length were significantly more negative. Treated kidneys were an additional 1.26 +/- 0.49 SD units below their expected length (p = 0.02). Untreated kidneys were further below normal as well but possibly to a lesser degree (-0.82 +/- 0.36; p <0.04). Although there was a trend for the abnormal group to have smaller kidneys than the normal group, both groups showed the same trend toward an age-adjusted reduction in renal growth at follow-up. The alterations in renal growth patterns observed in this population are unsettling and could be secondary to either treatment effect (SWL) or, more likely, to some underlying pathology intrinsic to pediatric kidneys with urolithiasis. Until further data are available, SWL in the pediatric population should be applied with caution and at the lowest dosage sufficient to achieve stone comminution. PMID- 9847073 TI - Literature watch. PMID- 9847074 TI - Toward a complete human genome sequence. AB - We have begun a joint program as part of a coordinated international effort to determine a complete human genome sequence. Our strategy is to map large-insert bacterial clones and to sequence each clone by a random shotgun approach followed by directed finishing. As of September 1998, we have identified the map positions of bacterial clones covering approximately 860 Mb for sequencing and completed >98 Mb ( approximately 3.3%) of the human genome sequence. Our progress and sequencing data can be accessed via the World Wide Web (http://webace.sanger.ac.uk/HGP/ or http://genome.wustl.edu/gsc/). PMID- 9847075 TI - WebWise: web sites of the Human Genome Project. PMID- 9847076 TI - Clusters of resistance genes in plants evolve by divergent selection and a birth and-death process. AB - Classical genetic and molecular data show that genes determining disease resistance in plants are frequently clustered in the genome. Genes for resistance (R genes) to diverse pathogens cloned from several species encode proteins that have motifs in common. These motifs indicate that R genes are part of signal transduction systems. Most of these R genes encode a leucine-rich repeat (LRR) region. Sequences encoding putative solvent-exposed residues in this region are hypervariable and have elevated ratios of nonsynonymous to synonymous substitutions; this suggests that they have evolved to detect variation in pathogen-derived ligands. Generation of new resistance specificities previously had been thought to involve frequent unequal crossing-over and gene conversions. However, comparisons between resistance haplotypes reveal that orthologs are more similar than paralogs implying a low rate of sequence homogenization from unequal crossing-over and gene conversion. We propose a new model adapted and expanded from one proposed for the evolution of vertebrate major histocompatibility complex and immunoglobulin gene families. Our model emphasizes divergent selection acting on arrays of solvent-exposed residues in the LRR resulting in evolution of individual R genes within a haplotype. Intergenic unequal crossing over and gene conversions are important but are not the primary mechanisms generating variation. PMID- 9847077 TI - Snapshot of a large dynamic replicon in a halophilic archaeon: megaplasmid or minichromosome? AB - Extremely halophilic archaea, which flourish in hypersaline environments, are known to contain a variety of large dynamic replicons. Previously, the analysis of one such replicon, pNRC100, in Halobacterium sp. strain NRC-1, showed that it undergoes high-frequency insertion sequence (IS) element-mediated insertions and deletions, as well as inversions via recombination between 39-kb-long inverted repeats (IRs). Now, the complete sequencing of pNRC100, a 191,346-bp circle, has shown the presence of 27 IS elements representing eight families. A total of 176 ORFs or likely genes of 850-bp average size were found, 39 of which were repeated within the large IRs. More than one-half of the ORFs are likely to represent novel genes that have no known homologs in the databases. Among ORFs with previously characterized homologs, three different copies of putative plasmid replication and four copies of partitioning genes were found, suggesting that pNRC100 evolved from IS element-mediated fusions of several smaller plasmids. Consistent with this idea, putative genes typically found on plasmids, including those encoding a restriction-modification system and arsenic resistance, as well as buoyant gas-filled vesicles and a two-component regulatory system, were found on pNRC100. However, additional putative genes not expected on an extrachromosomal element, such as those encoding an electron transport chain cytochrome d oxidase, DNA nucleotide synthesis enzymes thioredoxin and thioredoxin reductase, and eukaryotic-like TATA-binding protein transcription factors and a chromosomal replication initiator protein were also found. A multi step IS element-mediated process is proposed to account for the acquisition of these chromosomal genes. The finding of essential genes on pNRC100 and its property of resistance to curing suggest that this replicon may be evolving into a new chromosome. PMID- 9847078 TI - Mutation detection by ligation to complete n-mer DNA arrays. AB - A new approach to comparative nucleic acid sequence analysis is described that uses the ligation of DNA targets to high-density arrays containing complete sets of covalently attached oligonucleotides of length eight and nine. The combination of enzymatic or chemical ligation with a directed comparative analysis avoids many of the intrinsic difficulties associated with hybridization-based de novo sequence reconstruction methods described previously. Double-stranded DNA targets were fragmented and labeled to produce quasirandom populations of 5' termini suitable for ligation and detection on the arrays. Kilobase-size DNA targets were used to demonstrate that complete n-mer arrays can correctly verify known sequences and can determine the presence of sequence differences relative to a reference. By use of 9-mer arrays, sequences of 1.2-kb targets were verified with >99.9% accuracy. Mutations in target sequences were detected by directly comparing the intensity pattern obtained for an unknown with that obtained for a known reference sequence. For targets of moderate length (1.2 kb), 100% of the mutations in the queried sequences were detected with 9-mer arrays. For higher complexity targets (2.5 and 16.6 kb), a relatively high percentage of mutations (90% and 66%, respectively) were correctly identified with a low false-positive rate of <0.03 percent. The methods described provide a general approach to analyzing nucleic acid samples on the basis of the interpretation of sequence specific patterns of hybridization and ligation on complete n-mer oligonucleotide arrays. PMID- 9847079 TI - How to interpret an anonymous bacterial genome: machine learning approach to gene identification. AB - In this report we address the problem of accurate statistical modeling of DNA sequences, either coding or noncoding, for a bacterial species whose genome (or a large portion) was sequenced but not yet characterized experimentally. Availability of these models is critical for successful solution of the genome annotation task by statistical methods of gene finding. We present the method, GeneMark-Genesis, which learns the parameters of Markov models of protein-coding and noncoding regions from anonymous bacterial genomic sequence. These models are subsequently used in the GeneMark and GeneMark.hmm gene-finding programs. Although there is basically one model of a noncoding region for a given genome, several models of protein-coding region are automatically obtained by GeneMark Genesis. The diversity of protein-coding models reflects the diversity of oligonucleotide compositions, particularly the diversity of codon usage strategies observed in genes from one and the same genome. In the simplest and the most important case, there are just two gene models-typical and atypical ones. We show that the atypical model allows one to predict genes that escape identification by the typical model. Many genes predicted by the atypical model appear to be horizontally transferred genes. The early versions of GeneMark Genesis were used for annotating the genomes of Methanoccocus jannaschii and Helicobacter pylori. We report the results of accuracy testing of the full-scale version of GeneMark-Genesis on 10 completely sequenced bacterial genomes. Interestingly, the GeneMark.hmm program that employed the typical and atypical models defined by GeneMark-Genesis was able to predict 683 new atypical genes with 176 of them confirmed by similarity search. PMID- 9847082 TI - Predicting gene regulatory elements in silico on a genomic scale. AB - We performed a systematic analysis of gene upstream regions in the yeast genome for occurrences of regular expression-type patterns with the goal of identifying potential regulatory elements. To achieve this goal, we have developed a new sequence pattern discovery algorithm that searches exhaustively for a priori unknown regular expression-type patterns that are over-represented in a given set of sequences. We applied the algorithm in two cases, (1) discovery of patterns in the complete set of >6000 sequences taken upstream of the putative yeast genes and (2) discovery of patterns in the regions upstream of the genes with similar expression profiles. In the first case, we looked for patterns that occur more frequently in the gene upstream regions than in the genome overall. In the second case, first we clustered the upstream regions of all the genes by similarity of their expression profiles on the basis of publicly available gene expression data and then looked for sequence patterns that are over-represented in each cluster. In both cases we considered each pattern that occurred at least in some minimum number of sequences, and rated them on the basis of their over-representation. Among the highest rating patterns, most have matches to substrings in known yeast transcription factor-binding sites. Moreover, several of them are known to be relevant to the expression of the genes from the respective clusters. Experiments on simulated data show that the majority of the discovered patterns are not expected to occur by chance. PMID- 9847080 TI - Construction of an approximately 700-kb transcript map around the familial Mediterranean fever locus on human chromosome 16p13.3. AB - We used a combination of cDNA selection, exon amplification, and computational prediction from genomic sequence to isolate transcribed sequences from genomic DNA surrounding the familial Mediterranean fever (FMF) locus. Eighty-seven kb of genomic DNA around D16S3370, a marker showing a high degree of linkage disequilibrium with FMF, was sequenced to completion, and the sequence annotated. A transcript map reflecting the minimal number of genes encoded within the approximately 700 kb of genomic DNA surrounding the FMF locus was assembled. This map consists of 27 genes with discreet messages detectable on Northerns, in addition to three olfactory-receptor genes, a cluster of 18 tRNA genes, and two putative transcriptional units that have typical intron-exon splice junctions yet do not detect messages on Northerns. Four of the transcripts are identical to genes described previously, seven have been independently identified by the French FMF Consortium, and the others are novel. Six related zinc-finger genes, a cluster of tRNAs, and three olfactory receptors account for the majority of transcribed sequences isolated from a 315-kb FMF central region (between D16S468/D16S3070 and cosmid 377A12). Interspersed among them are several genes that may be important in inflammation. This transcript map not only has permitted the identification of the FMF gene (MEFV), but also has provided us an opportunity to probe the structural and functional features of this region of chromosome 16. PMID- 9847081 TI - CeRep25B forms chromosome-specific minisatellite arrays in Caenorhabditis elegans. AB - With the completion of the Genome Sequencing Project, it is now possible to rapidly and accurately determine the frequency and position of a particular repeat sequence in the Caenorhabditis elegans genome. Several repeat sequences with a variety of characteristics have been examined and with few exceptions they show a near-random distribution throughout the genome. We characterized several genes near the left end of Chromosome III in the C. elegans genome, and found a 24-bp minisatellite repeat sequence present in the introns of two unrelated genes. This prompted a search of the databank for other occurrences of this sequence. Multiple copy arrays of this repeat are all located on the same autosome and fall in two clusters: one near the left end, and one in the central region separated by approximately 10 Mb. There are >200 copies of this repeat on the chromosome. This euchromatic repeat sequence seems unrelated to gene expression, is absent from homologous sites in a related species, is unstable in Escherichia coli, and is polymorphic between different wild isolates of C. elegans. Most CeRep25B units in the array match the consensus sequence very well, suggesting that either this repeat originated quite recently or its sequence is functionally constrained. Although chromosome-specific repeat sequences have been reported previously in many organisms, such sequences are usually structural and heterochromatic (e.g., centromeric alpha-satellite) or on the mammalian sex chromosomes. This report describes the first confirmed instance from a whole genome sequencing project of an autosomal euchromatic chromosome-specific minisatellite repeat. PMID- 9847084 TI - Deepwater rice: A model plant to study stem elongation PMID- 9847085 TI - Transcriptional regulation in plants: the importance of combinatorial control. PMID- 9847083 TI - Quality assessment of whole genome mapping data in the refined familial spastic paraplegia interval on chromosome 14q. AB - Autosomal dominant familial spastic paraplegia (AD-FSP) is a genetically heterogeneous neurodegenerative disorder characterized by progressive spasticity of the lower limbs. Three loci on chromosome 14q (SPG3), 2p (SPG4), and 15q (SPG6) were shown to be responsible for AD-FSP. Analysis of recombination events in three SPG3-linked families allowed us to narrow the critical interval from 9 to 5 cM. An approximately 5-Mb YAC contig comprising 32 clones and 90 STSs was built from D14S301 to D14S991, encompassing this region of 14q21. Fifty-six ESTs assigned previously to this region with radiation hybrid (RH) panels Genebridge 4 and G3 were precisely localized on the YAC contig. The 90 STSs positioned on the contig were tested on the TNG RH panel to compare our YAC-based map with an RH map at a high level of resolution. Comparison between our map and the whole genome mapping data on this interval of chromosome 14q is discussed. PMID- 9847086 TI - Plant hemoglobins PMID- 9847087 TI - 3-Methylcrotonyl-coenzyme A carboxylase is a component of the mitochondrial leucine catabolic pathway in plants AB - 3-Methylcrotonyl-coenzyme A carboxylase (MCCase) is a mitochondrial biotin containing enzyme whose metabolic function is not well understood in plants. In soybean (Glycine max) seedlings the organ-specific and developmentally induced changes in MCCase expression are regulated by mechanisms that control the accumulation of MCCase mRNA and the activity of the enzyme. During soybean cotyledon development, when seed-storage proteins are degraded, leucine (Leu) accumulation peaks transiently at 8 d after planting. The coincidence between peak MCCase expression and the decline in Leu content provides correlative evidence that MCCase is involved in the mitochondrial catabolism of Leu. Direct evidence for this conclusion was obtained from radiotracer metabolic studies using extracts from isolated mitochondria. These experiments traced the metabolic fate of [U-14C]Leu and NaH14CO3, the latter of which was incorporated into methylglutaconyl-coenzyme A (CoA) via MCCase. These studies directly demonstrate that plant mitochondria can catabolize Leu via the following scheme: Leu --> alpha-ketoisocaproate --> isovaleryl-CoA --> 3-methylcrotonyl-CoA --> 3 methylglutaconyl-CoA --> 3-hydroxy-3-methylglutaryl-CoA --> acetoacetate + acetyl CoA. These findings demonstrate for the first time, to our knowledge, that the enzymes responsible for Leu catabolism are present in plant mitochondria. We conclude that a primary metabolic role of MCCase in plants is the catabolism of Leu. PMID- 9847088 TI - Root-growth behavior of the Arabidopsis mutant rgr1. Roles of gravitropism and circumnutation in the waving/coiling phenomenon. AB - In this study we investigated the kinetics of the gravitropic response of the Arabidopsis mutant rgr1 (reduced root gravitropism). Although the rate of curvature in rgr1, which is allelic to axr4, was smaller than in the wild type (ecotype Wassilewskija), curvature was initiated in the same region of the root, the distal elongation zone. The time lag for the response was unaffected in the mutant; however, the gravitropic response of rgr1 contained a feature not found in the wild type: when roots growing along the surface of an agar plate were gravistimulated, there was often an upward curvature that initiated in the central elongation zone. Because this response was dependent on the tactile environment of the root, it most likely resulted from the superposition of the waving/coiling phenomenon onto the gravitropic response. We found that the frequency of the waving pattern and circumnutation, a cyclic endogenous pattern of root growth, was the same in rgr1 and in the wild type, so the waving/coiling phenomenon is likely governed by circumnutation patterns. The amplitudes of these oscillations may then be selectively amplified by tactile stimulation to provide a directional preference to the slanting. PMID- 9847089 TI - Characterization of a low-molecular-weight glutenin subunit gene from bread wheat and the corresponding protein that represents a major subunit of the glutenin polymer. AB - Both high- and low-molecular-weight glutenin subunits (LMW-GS) play the major role in determining the viscoelastic properties of wheat (Triticum aestivum L.) flour. To date there has been no clear correspondence between the amino acid sequences of LMW-GS derived from DNA sequencing and those of actual LMW-GS present in the endosperm. We have characterized a particular LMW-GS from hexaploid bread wheat, a major component of the glutenin polymer, which we call the 42K LMW-GS, and have isolated and sequenced the putative corresponding gene. Extensive amino acid sequences obtained directly for this 42K LMW-GS indicate correspondence between this protein and the putative corresponding gene. This subunit did not show a cysteine (Cys) at position 5, in contrast to what has frequently been reported for nucleotide-based sequences of LMW-GS. This Cys has been replaced by one occurring in the repeated-sequence domain, leaving the total number of Cys residues in the molecule the same as in various other LMW-GS. On the basis of the deduced amino acid sequence and literature-based assignment of disulfide linkages, a computer-generated molecular model of the 42K subunit was constructed. PMID- 9847090 TI - Chlorophyll synthesis in dark-grown pine primary needles AB - The pigment content of dark-grown primary needles of Pinus jeffreyi L. and Pinus sylvestris L. was determined by high-performance liquid chromatography. The state of protochlorophyllide a and of chlorophylls during dark growth were analyzed by in situ 77 K fluorescence spectroscopy. Both measurements unambiguously demonstrated that pine primary needles are able to synthesize chlorophyll in the dark. Norflurazon strongly inhibited both carotenoid and chlorophyll synthesis. Needles of plants treated with this inhibitor had low chlorophyll content, contained only traces of xanthophylls, and accumulated carotenoid precursors. The first form of chlorophyll detected in young pine needles grown in darkness had an emission maximum at 678 nm. Chlorophyll-protein complexes with in situ spectroscopic properties similar to those of fully green needles (685, 695, and 735 nm) later accumulated in untreated plants, whereas in norflurazon-treated plants the photosystem I emission at 735 nm was completely lacking. To better characterize the light-dependent chlorophyll biosynthetic pathway in pine needles, the 77 K fluorescence properties of in situ protochlorophyllide a spectral forms were studied. Photoactive and nonphotoactive protochlorophyllide a forms with emission properties similar to those reported for dark-grown angiosperms were found, but excitation spectra were substantially red shifted. Because of their lower chlorophyll content, norflurazon-treated plants were used to study the protochlorophyllide a photoreduction process triggered by one light flash. The first stable chlorophyllide photoproduct was a chlorophyllide a form emitting at 688 nm as in angiosperms. Further chlorophyllide a shifts usually observed in angiosperms were not detected. The rapid regeneration of photoactive protochlorophyllide a from nonphotoactive protochlorophyllide after one flash was demonstrated. PMID- 9847092 TI - Abscisic acid-dependent and -independent expression of the carrot late embryogenesis-abundant-class gene Dc3 in transgenic tobacco seedlings AB - We studied the expression of three promoter 5' deletion constructs (-218, -599, and -1312) of the LEA (late embryogenesis abundant)-class gene Dc3 fused to beta glucuronidase (GUS), where each construct value refers to the number of base pairs upstream of the transcription start site at which the deletion occurred. The Dc3 gene is noted for its induction by abscisic acid (ABA), but its response to other plant hormones and various environmental stresses has not been reported previously for vegetative cells. Fourteen-day-old transgenic tobacco (Nicotiana tabacum L.) seedlings were exposed to dehydration, hypoxia, salinity, exogenous ethylene, or exogenous methyl jasmonate (MeJa). GUS activity was quantified fluorimetrically and expression was observed by histochemical staining of the seedlings. An increase in GUS activity was observed in plants with constructs 599 and -1312 in response to dehydration and salinity within 6 h of stress, and at 12 h in response to hypoxia. No increase in endogenous ABA was found in any of the three lines, even after 72 h of hypoxia. An ABA-independent increase in GUS activity was observed when endogenous ABA biosynthesis was blocked by fluridone and plants were exposed to 5 &mgr;L L-1 ethylene in air or 100 &mgr;M MeJa. Virtually no expression was observed in construct -218 in response to dehydration, salinity, or MeJa, but there was a moderate response to ethylene and hypoxia. This suggests that the region between -218 and -599 is necessary for ABA (dehydration and salinity)- and MeJa-dependent expression, whereas ethylene mediated expression does not require this region of the promoter. PMID- 9847091 TI - Nucellain, a barley homolog of the dicot vacuolar-processing protease, is localized in nucellar cell walls. AB - The nucellus is a complex maternal grain tissue that embeds and feeds the developing cereal endosperm and embryo. Differential screening of a barley (Hordeum vulgare) cDNA library from 5-d-old ovaries resulted in the isolation of two cDNA clones encoding nucellus-specific homologs of the vacuolar-processing enzyme of castor bean (Ricinus communis). Based on the sequence of these barley clones, which are called nucellains, a homolog from developing corn (Zea mays) grains was also identified. In dicots the vacuolar-processing enzyme is believed to be involved in the processing of vacuolar storage proteins. RNA-blot and in situ-hybridization analyses detected nucellain transcripts in autolysing nucellus parenchyma cells, in the nucellar projection, and in the nucellar epidermis. No nucellain transcripts were detected in the highly vacuolate endosperm or in the other maternal tissues of developing grains such as the testa or the pericarp. Using an antibody raised against castor bean vacuolar-processing protease, a single polypeptide was recognized in protein extracts from barley grains. Immunogold-labeling experiments with this antibody localized the nucellain epitope not in the vacuoles, but in the cell walls of all nucellar cell types. We propose that nucellain plays a role in processing and/or turnover of cell wall proteins in developing cereal grains. PMID- 9847093 TI - Quantitative control of inflorescence formation in impatiens balsamina AB - We analyzed the process of inflorescence formation in Impatiens balsamina by studying the architecture of the plant under different photoperiod treatments. Floral reversion under noninductive conditions in this species is caused by the lack of persistence of the induced state in the leaf. This can be used to control the amount of inductive signal and to examine its quantitative influence on morphological changes in the plant. The floral transition was characterized by a continuum of variation at the level of meristem identity, primordium initiation, and floral organ identity. This continuum was enhanced during reversion, suggesting that the establishment of a continuum partly reflects limiting amounts of inductive signal exported from the leaf to the meristem. The transcription patterns of two homologs of genes involved in the control of floral meristem identity, Imp-FLO and Imp-FIM, were similar in terminal and axillary flowers and may be associated with the continuum exhibited by I. balsamina. By analyzing the fate of axillary meristem primordia initiated before and after the beginning of the inductive period, we showed that de novo initiation of axillary meristem primordia by the evoked meristem is not required and that primordia initiated before evocation can adopt different fates, depending on the amount of inductive signal. The influence of age and/or position on primordium responsiveness to the inductive signal is discussed. PMID- 9847094 TI - Benzothiadiazole-mediated induced resistance to fusarium oxysporum f. sp. radicis lycopersici in tomato AB - Benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), a synthetic chemical, was applied as a foliar spray to tomato (Lycopersicon esculentum) plants and evaluated for its potential to confer increased resistance against the soil-borne pathogen Fusarium oxysporum f. sp. radicis-lycopersici (FORL). In nontreated tomato plants all root tissues were massively colonized by FORL hyphae. Pathogen ingress toward the vascular stele was accompanied by severe host cell alterations, including cell wall breakdown. In BTH-treated plants striking differences in the rate and extent of fungal colonization were observed. Pathogen growth was restricted to the epidermis and the outer cortex, and fungal ingress was apparently halted by the formation of callose-enriched wall appositions at sites of fungal penetration. In addition, aggregated deposits, which frequently established close contact with the invading hyphae, accumulated in densely colonized epidermal cells and filled most intercellular spaces. Upon incubation of sections with gold-complexed laccase for localization of phenolic-like compounds, a slight deposition of gold particles was observed over both the host cell walls and the wall appositions. Labeling was also detected over the walls of fungal cells showing signs of obvious alteration ranging from cytoplasm disorganization to protoplasm retraction. We provide evidence that foliar applications of BTH sensitize susceptible tomato plants to react more rapidly and more efficiently to FORL attack through the formation of protective layers at sites of potential fungal entry. PMID- 9847095 TI - Spermine is a salicylate-independent endogenous inducer for both tobacco acidic pathogenesis-related proteins and resistance against tobacco mosaic virus infection AB - Intercellular spaces are often the first sites invaded by pathogens. In the spaces of tobacco mosaic virus (TMV)-infected and necrotic lesion-forming tobacco (Nicotiana tabacum L.) leaves, we found that an inducer for acidic pathogenesis related (PR) proteins was accumulated. The induction activity was recovered in gel-filtrated fractions of low molecular mass with a basic nature, into which authentic spermine (Spm) was eluted. We quantified polyamines in the intercellular spaces of the necrotic lesion-forming leaves and found 20-fold higher levels of free Spm than in healthy leaves. Among several polyamines tested, exogenously supplied Spm induced acidic PR-1 gene expression. Immunoblot analysis showed that Spm treatment increased not only acidic PR-1 but also acidic PR-2, PR-3, and PR-5 protein accumulation. Treatment of healthy tobacco leaves with salicylic acid (SA) caused no significant increase in the level of endogenous Spm, and Spm did not increase the level of endogenous SA, suggesting that induction of acidic PR proteins by Spm is independent of SA. The size of TMV induced local lesions was reduced by Spm treatment. These results indicate that Spm accumulates outside of cells after lesion formation and induces both acidic PR proteins and resistance against TMV via a SA-independent signaling pathway. PMID- 9847096 TI - Meristem-specific suppression of mitosis and a global switch in gene expression in the root cap of pea by endogenous signals AB - Two functionally distinct sets of meristematic cells exist within root tips of pea (Pisum sativum): the root apical meristem, which gives rise to the body of the root; and the root cap meristem, which gives rise to cells that differentiate progressively through the cap and separate ultimately from its periphery as border cells. When a specific number of border cells has accumulated on the root cap periphery, mitosis within the root cap meristem, but not the apical meristem, is suppressed. When border cells are removed by immersion of the root tip in water, a transient induction of mitosis in the root cap meristem can be detected starting within 5 min. A corresponding switch in gene expression throughout the root cap occurs in parallel with the increase in mitosis, and new border cells begin to separate from the root cap periphery within 1 h. The induction of renewed border cell production is inhibited by incubating root tips in extracellular material released from border cells. The results are consistent with the hypothesis that operation of the root cap meristem and consequent turnover of the root cap is self-regulated by a signal from border cells. PMID- 9847097 TI - A gene encoding proline dehydrogenase is not only induced by proline and hypoosmolarity, but is also developmentally regulated in the reproductive organs of Arabidopsis. AB - The cDNA clone ERD5 (early responsive to dehydration), isolated from 1-h dehydrated Arabidopsis, encodes a precursor of proline (Pro) dehydrogenase (ProDH), which is a mitochondrial enzyme involved in the first step of the conversion of Pro to glutamic acid. The transcript of the erd5 (ProDH) gene was undetectable when plants were dehydrated, but large amounts of transcript accumulated when plants were subsequently rehydrated. Accumulation of the transcript was also observed in plants that had been incubated under hypoosmotic conditions in media that contained L- or D-Pro. We isolated a 1.4-kb DNA fragment of the putative promoter region of the ProDH gene. The beta-glucuronidase (GUS) reporter gene driven by the 1.4-kb ProDH promoter was induced not only by rehydration but also by hypoosmolarity and L- and D-Pro at significant levels in transgenic Arabidopsis plants. The promoter of the ProDH gene directs strong GUS activity in reproductive organs such as pollen and pistils and in the seeds of the transgenic plants. GUS activity was detected in vegetative tissues such as veins of leaves and root tips when the transgenic plants were exposed to hypoosmolarity and Pro solutions. GUS activity increased during germination of the transgenic plants under hypoosmolarity. The relationship between Pro metabolism and the physiological aspects of stress response and development are discussed. PMID- 9847098 TI - Ozone sensitivity in hybrid poplar is correlated with a lack of defense-gene activation AB - Ozone is a major gaseous pollutant thought to contribute to forest decline. Although the physiological and morphological responses of forest trees to ozone have been well characterized, little is known about the molecular basis for these responses. Our studies compared the response to ozone of ozone-sensitive and ozone-tolerant clones of hybrid poplar (Populus maximowizii x Populus trichocarpa) at the physiological and molecular levels. Gas-exchange analyses demonstrated clear differences between the ozone-sensitive clone 388 and the ozone-tolerant clone 245. Although ozone induced a decrease in photosynthetic rate and stomatal conductance in both clones, the magnitude of the decrease in stomatal conductance was significantly greater in the ozone-tolerant clone. RNA blot analysis established that ozone-induced mRNA levels for phenylalanine ammonia-lyase, O-methyltransferase, a pathogenesis-related protein, and a wound inducible gene were significantly higher in the ozone-tolerant than in the ozone sensitive plants. Wound- and pathogen-induced levels of these mRNAs were also higher in the ozone-tolerant compared with the ozone-sensitive plants. The different physiological and molecular responses to ozone exposure exhibited by clones 245 and 388 suggest that ozone tolerance involves the activation of salicylic-acid- and jasmonic-acid-mediated signaling pathways, which may be important in triggering defense responses against oxidative stress. PMID- 9847099 TI - The responses of cytochrome redox state and energy metabolism to dehydration support a role for cytoplasmic viscosity in desiccation tolerance AB - To characterize the depression of metabolism in anhydrobiotes, the redox state of cytochromes and energy metabolism were studied during dehydration of soaked cowpea (Vigna unguiculata) cotyledons and pollens of Typha latifolia and Impatiens glandulifera. Between water contents (WC) of 1.0 and 0.6 g H2O/g dry weight (g/g), viscosity as measured by electron spin resonance spectroscopy increased from 0.15 to 0.27 poise. This initial water loss was accompanied by a 50% decrease in respiration rates, whereas the adenylate energy charge remained constant at 0.8, and cytochrome c oxidase (COX) remained fully oxidized. From WC of 0.6 to 0.2 g/g, viscosity increased exponentially. The adenylate energy charge declined to 0.4 in seeds and 0.2 in pollen, whereas COX became progressively reduced. At WC of less than 0.2 g/g, COX remained fully reduced, whereas respiration ceased. When dried under N2, COX remained 63% reduced in cotyledons until WC was 0.7 g/g and was fully reduced at 0.2 g/g. During drying under pure O2, the pattern of COX reduction was similar to that of air-dried tissues, although the maximum reduction was 70% in dried tissues. Thus, at WC of less than 0.6 g/g, the reduction of COX probably originates from a decreased O2 availability as a result of the increased viscosity and impeded diffusion. We suggest that viscosity is a valuable parameter to characterize the relation between desiccation and decrease in metabolism. The implications for desiccation tolerance are discussed. PMID- 9847101 TI - Developmental and environmental effects on the expression of the C3-C4 intermediate phenotype in moricandia arvensis AB - Cellular anatomy and expression of glycine decarboxylase (GDC) protein were studied during leaf development of the C3-C4 intermediate species Moricandia arvensis. Leaf anatomy was initially C3-like and the number and profile area of mitochondria in the bundle-sheath cells were the same as those in adjacent mesophyll cells. Between a leaf length of 6 and 12 mm there was a bundle-sheath specific, 4-fold increase in the number of mitochondrial profiles, followed by a doubling of their individual profile areas as the leaves expanded further. Subunits of GDC were present in whole-leaf extracts before the anatomical development of bundle-sheath cells. Whereas the GDC H-protein content of leaves increased steadily throughout development, the increase in GDC P-protein was synchronous with the development of mitochondria in the bundle sheath. The P protein was confined to bundle-sheath mitochondria throughout leaf development, and its content in individual mitochondria increased before the anatomical development of the bundle sheath. Anatomical and biochemical attributes of the C3 C4 character were present in the cotyledons and sepals but not in other photosynthetic organs/tissues. In leaves and cotyledons that developed in the dark, the expression of the P-protein and the organellar development were reduced but the bundle-sheath cell specificity was retained. PMID- 9847100 TI - NPH4, a conditional modulator of auxin-dependent differential growth responses in Arabidopsis. AB - Although sessile in nature, plants are able to use a number of mechanisms to modify their morphology in response to changing environmental conditions. Differential growth is one such mechanism. Despite its importance in plant development, little is known about the molecular events regulating the establishment of differential growth. Here we report analyses of the nph4 (nonphototropic hypocotyl) mutants of Arabidopsis that suggest that the NPH4 protein plays a central role in the modulation of auxin-dependent differential growth. Results from physiological studies demonstrate that NPH4 activity is conditionally required for a number of differential growth responses, including phototropism, gravitropism, phytochrome-dependent hypocotyl curvature, apical hook maintenance, and abaxial/adaxial leaf-blade expansion. The nph4 mutants exhibited auxin resistance and severely impaired auxin-dependent gene expression, indicating that the defects associated with differential growth likely arise because of altered auxin responsiveness. Moreover, the auxin signaling events mediating phototropism are genetically correlated with the abundance of the NPH4 protein. PMID- 9847102 TI - Differential regulation of enolase during anaerobiosis in maize. AB - It was reported previously that enolase enzyme activity and ENO1 transcript levels are induced by anaerobic stress in maize (Zea mays). Here we show that not all isoforms of maize enolase are anaerobically induced. We cloned and sequenced a second enolase cDNA clone (pENO2) from maize. Sequence analysis showed that pENO2 shares 75.6% nucleotide and 89.5% deduced amino acid sequence identity with pENO1 and is encoded by a distinct gene. Expression of ENO2 is constitutive under aerobic conditions, whereas ENO1 levels are induced 10-fold in maize roots after 24 h of anaerobic treatment. Western-blot analysis and N-terminal sequencing of in vivo-labeled maize roots identified two major proteins selectively synthesized upon anaerobic stress as isozymes of enolase. We describe the expression of enolase in maize roots under anaerobic stress. PMID- 9847103 TI - Differential expression and internal feedback regulation of 1-aminocyclopropane-1 carboxylate synthase, 1-aminocyclopropane-1-carboxylate oxidase, and ethylene receptor genes in tomato fruit during development and ripening. AB - We investigated the feedback regulation of ethylene biosynthesis in tomato (Lycopersicon esculentum) fruit with respect to the transition from system 1 to system 2 ethylene production. The abundance of LE-ACS2, LE-ACS4, and NR mRNAs increased in the ripening fruit concomitant with a burst in ethylene production. These increases in mRNAs with ripening were prevented to a large extent by treatment with 1-methylcyclopropene (MCP), an ethylene action inhibitor. Transcripts for the LE-ACS6 gene, which accumulated in preclimacteric fruit but not in untreated ripening fruit, did accumulate in ripening fruit treated with MCP. Treatment of young fruit with propylene prevented the accumulation of transcripts for this gene. LE-ACS1A, LE-ACS3, and TAE1 genes were expressed constitutively in the fruit throughout development and ripening irrespective of whether the fruit was treated with MCP or propylene. The transcripts for LE-ACO1 and LE-ACO4 genes already existed in preclimacteric fruit and increased greatly when ripening commenced. These increases in LE-ACO mRNA with ripening were also prevented by treatment with MCP. The results suggest that in tomato fruit the preclimacteric system 1 ethylene is possibly mediated via constitutively expressed LE-ACS1A and LE-ACS3 and negatively feedback-regulated LE-ACS6 genes with preexisting LE-ACO1 and LE-ACO4 mRNAs. At the onset of the climacteric stage, it shifts to system 2 ethylene, with a large accumulation of LE-ACS2, LE ACS4, LE-ACO1, and LE-ACO4 mRNAs as a result of a positive feedback regulation. This transition from system 1 to system 2 ethylene production might be related to the accumulated level of NR mRNA. PMID- 9847104 TI - Expression analysis of a ripening-specific, auxin-repressed endo-1, 4-beta glucanase gene in strawberry. AB - A cDNA (Cel1) encoding an endo-1,4-beta-glucanase (EGase) was isolated from ripe fruit of strawberry (Fragaria x ananassa). The deduced protein of 496 amino acids contains a presumptive signal sequence, a common feature of cell wall-localized EGases, and one potential N-glycosylation site. Southern- blot analysis of genomic DNA from F. x ananassa, an octoploid species, and that from the diploid species Fragaria vesca indicated that the Cel1 gene is a member of a divergent multigene family. In fruit, Cel1 mRNA was first detected at the white stage of development, and at the onset of ripening, coincident with anthocyanin accumulation, Cel1 mRNA abundance increased dramatically and remained high throughout ripening and subsequent fruit deterioration. In all other tissues examined, Cel1 expression was invariably absent. Antibodies raised to Cel1 protein detected a protein of 62 kD only in ripening fruit. Upon deachenation of young white fruit to remove the source of endogenous auxins, ripening, as visualized by anthocyanin accumulation, and Cel1 mRNA accumulation were both accelerated. Conversely, auxin treatment of white fruit repressed accumulation of both Cel1 mRNA and ripening. These results indicate that strawberry Cel1 is a ripening-specific and auxin-repressed EGase, which is regulated during ripening by a decline in auxin levels originating from the achenes. PMID- 9847105 TI - Comparison of binding properties and early biological effects of elicitins in tobacco cells AB - Elicitins are a family of small proteins secreted by Phytophthora species that have a high degree of homology and elicit defense reactions in tobacco (Nicotiana tabacum). They display acidic or basic characteristics, the acidic elicitins being less efficient in inducing plant necrosis. In this study we compared the binding properties of four elicitins (two basic and two acidic) and early-induced signal transduction events (Ca2+ influx, extracellular medium alkalinization, and active oxygen species production). The affinity for tobacco plasma membrane binding sites and the number of binding sites were similar for all four elicitins. Furthermore, elicitins compete with one another for binding sites, suggesting that they interact with the same receptor. The four elicitins induced Ca2+ influx, extracellular medium alkalinization, and the production of active oxygen species in tobacco cell suspensions, but the intensity and kinetics of these effects were different from one elicitin to another. As a general observation the concentrations that induce similar levels of biological activities were lower for basic elicitins (with the exception of cinnamomin induced Ca2+ uptake). The qualitative similarity of early events induced by elicitins indicates a common transduction scheme, whereas fine signal transduction tuning is different in each elicitin. PMID- 9847107 TI - Distribution of sulfur within oilseed rape leaves in response to sulfur deficiency during vegetative growth AB - The distribution of S to sulfate, glucosinolates, glutathione, and the insoluble fraction within oilseed rape (Brassica napus L.) leaves of different ages was investigated during vegetative growth. The concentrations of glutathione and glucosinolates increased from the oldest to the youngest leaves, whereas the opposite was observed for SO42-. The concentration of insoluble S was similar among all of the leaves. At sufficient S supply and in the youngest leaves, 2% of total S was allocated to glutathione, 6% to glucosinolates, 50% to the insoluble fraction, and the remainder accumulated as SO42-. In the middle and oldest leaves, 70% to 90% of total S accumulated as SO42-, whereas glutathione and glucosinolates together accounted for less than 1% of S. When the S supply was withdrawn (minus S), the concentrations of all S-containing compounds, particularly SO42-, decreased in the youngest and middle leaves. Neither glucosinolates nor glutathione were major sources of S during S deficiency. Plants grown on nutrient solution containing minus S and low N were less deficient than plants grown on solution containing minus S and high N. The effect of N was explained by differences in growth rate. The different responses of leaves of different ages to S deficiency have to be taken into account for the development of field diagnostic tests to determine whether plants are S deficient. PMID- 9847106 TI - Role of the ascorbate-glutathione cycle of mitochondria and peroxisomes in the senescence of pea leaves AB - We investigated the relationship between H2O2 metabolism and the senescence process using soluble fractions, mitochondria, and peroxisomes from senescent pea (Pisum sativum L.) leaves. After 11 d of senescence the activities of Mn superoxide dismutase, dehydroascorbate reductase (DHAR), and glutathione reductase (GR) present in the matrix, and ascorbate peroxidase (APX) and monodehydroascorbate reductase (MDHAR) activities localized in the mitochondrial membrane, were all substantially decreased in mitochondria. The mitochondrial ascorbate and dehydroascorbate pools were reduced, whereas the oxidized glutathione levels were maintained. In senescent leaves the H2O2 content in isolated mitochondria and the NADH- and succinate-dependent production of superoxide (O2.-) radicals by submitochondrial particles increased significantly. However, in peroxisomes from senescent leaves both membrane-bound APX and MDHAR activities were reduced. In the matrix the DHAR activity was enhanced and the GR activity remained unchanged. As a result of senescence, the reduced and the oxidized glutathione pools were considerably increased in peroxisomes. A large increase in the glutathione pool and DHAR activity were also found in soluble fractions of senescent pea leaves, together with a decrease in GR, APX, and MDHAR activities. The differential response to senescence of the mitochondrial and peroxisomal ascorbate-glutathione cycle suggests that mitochondria could be affected by oxidative damage earlier than peroxisomes, which may participate in the cellular oxidative mechanism of leaf senescence longer than mitochondria. PMID- 9847108 TI - 12-Oxophytodienoate-10,11-reductase: occurrence of two isoenzymes of different specificity against stereoisomers of 12-oxophytodienoic acid AB - The reduction of 12-oxophytodienoic acid (OPDA) to 3-oxo-2(2'[Z]-pentenyl) cyclopentane-1-octanoic acid is catalyzed by 12-oxophytodienoate-10,11-reductase (OPR). Analysis of the isomer preference of OPR has indicated that the activity is composed of two isoenzymes exhibiting different stereoselectivities. The two isoforms of OPR have been separated, using protein extracts of Rock Harlequin (Corydalis sempervirens) as the starting material. OPRI, the enzyme reported earlier from the same species and corresponding to the cloned OPR from Arabidopsis, utilized 9R,13R-OPDA >> 9S, 13R-OPDA but not the 13S-configured isomers, whereas the new activity, OPRII, effectively reduced all four OPDA isomers, including the natural 9S,13S-OPDA (cis-[+]-OPDA). OPRII activity is characterized in detail. The enzyme's enzymatic, biochemical, and immunological properties prove that it is a close relative of OPRI. The roles of OPRI and OPRII in octadecanoid biology are discussed. PMID- 9847109 TI - Comparison of the ability of partially N-acetylated chitosans and chitooligosaccharides to elicit resistance reactions in wheat leaves AB - Chitin, a linear polysaccharide composed of (1-->4)-linked 2-acetamido-2-deoxy beta-D-glucopyranose (GlcNAc) residues, and chitosan, the fully or partially N acetylated, water-soluble derivative of chitin composed of (1-->4)-linked GlcNAc and 2-amino-2-deoxy-beta-D-glucopyranose (GlcN), have been proposed as elicitors of defense reactions in higher plants. We tested and compared the ability of purified (1-->4)-linked oligomers of GlcNAc (tetramer to decamer) and of GlcN (pentamer and heptamer) and partially N-acetylated chitosans with degrees of acetylation (DA) of 1%, 15%, 35%, 49%, and 60% and average degrees of polymerization between 540 and 1100 to elicit phenylalanine ammonia-lyase (PAL) and peroxidase (POD) activities, lignin deposition, and microscopically and macroscopically visible necroses when injected into the intercellular spaces of healthy, nonwounded wheat (Triticum aestivum L.) leaves. Purified oligomers of (1 ->4)-linked GlcN were not active as elicitors, whereas purified oligomers of (1- >4)-linked GlcNAc with a degree of polymerization >/= 7 strongly elicited POD activities but not PAL activities. Partially N-acetylated, polymeric chitosans elicited both PAL and POD activities, and maximum elicitation was observed with chitosans of intermediate DAs. All chitosans but not the chitin oligomers induced the deposition of lignin, the appearance of necrotic cells exhibiting yellow autofluorescence under ultraviolet light, and macroscopically visible necroses; those with intermediate DAs were most active. These results suggest that different mechanisms are involved in the elicitation of POD activities by GlcNAc oligomers, and of PAL and POD activities by partially N-acetylated chitosan polymers and that both enzymes have to be activated for lignin biosynthesis and ensuing necrosis to occur. PMID- 9847110 TI - Regulation of apoplastic NH4+ concentration in leaves of oilseed rape AB - Regulation of apoplastic NH4+ concentration in leaves of oilseed rape (Brassica napus L.) was studied using a vacuum-infiltration technique that allowed controlled manipulations of the apoplastic solution. In leaves infiltrated with NH4+-free solution, the apoplastic NH4+ concentration returned in less than 1.5 min to the preinfiltration level of 0.8 mM. Infiltrated 15NH4+ was rapidly diluted by 14NH4+/14NH3 effluxed from the cell. The exchange rate of 15N/14N over the apoplast due to combined 14N efflux from the symplast and 15N influx from the apoplastic solution was 29.4 &mgr;mol g-1 fresh weight h-1 between 0 and 5 min after infiltration. The net uptake of NH4+ into the leaf cells increased linearly with apoplastic NH4+ concentrations between 2 and 10 mM and could be partially inhibited by the channel inhibitors La3+ and tetraethylammonium and by Na+ and K+. When apoplastic pH increased from 5.0 to 8.0, the steady-state apoplastic NH4+ concentration decreased from 1.0 to 0.3 mM. Increasing temperature increased the rate of NH4+ net uptake and reduced the apoplastic steady-state NH4+ concentration. We conclude that the apoplastic solution in leaves of oilseed rape constitutes a highly dynamic NH4+ pool. PMID- 9847111 TI - Inhibition of auxin movement from the shoot into the root inhibits lateral root development in Arabidopsis. AB - In roots two distinct polar movements of auxin have been reported that may control different developmental and growth events. To test the hypothesis that auxin derived from the shoot and transported toward the root controls lateral root development, the two polarities of auxin transport were uncoupled in Arabidopsis. Local application of the auxin-transport inhibitor naphthylphthalamic acid (NPA) at the root-shoot junction decreased the number and density of lateral roots and reduced the free indoleacetic acid (IAA) levels in the root and [3H]IAA transport into the root. Application of NPA to the basal half of or at several positions along the root only reduced lateral root density in regions that were in contact with NPA or in regions apical to the site of application. Lateral root development was restored by application of IAA apical to NPA application. Lateral root development in Arabidopsis roots was also inhibited by excision of the shoot or dark growth and this inhibition was reversible by IAA. Together, these results are consistent with auxin transport from the shoot into the root controlling lateral root development. PMID- 9847112 TI - The respiratory burst and electrolyte leakage induced by sulfhydryl blockers in egeria densa leaves are associated with H2O2 production and are dependent on Ca2+ influx AB - In leaves of Egeria densa Planchon, N-ethylmaleimide (NEM) and other sulfhydryl binding reagents induce a temporary increase in nonmitochondrial respiration (DeltaQO2) that is inhibited by diphenylene iodonium and quinacrine, two known inhibitors of the plasma membrane NADPH oxidase, and are associated with a relevant increase in electrolyte leakage (M. Bellando, S. Sacco, F. Albergoni, P. Rocco, M.T. Marre [1997] Bot Acta 110: 388-394). In this paper we report data indicating further analogies between the oxidative burst induced by sulfhydryl blockers in E. densa and that induced by pathogen-derived elicitors in animal and plant cells: (a) NEM- and Ag+-induced DeltaQO2 was associated with H2O2 production and both effects depended on the presence of external Ca2+; (b) Ca2+ influx was markedly increased by treatment with NEM; (c) the Ca2+ channel blocker LaCl3 inhibited DeltaQO2, electrolyte release, and membrane depolarization induced by the sulfhydryl reagents; and (d) LaCl3 also inhibited electrolyte leakage induced by the direct infiltration of the leaves with H2O2. These results suggest a model in which the interaction of sulfhydryl blockers with sulfhydryl groups of cell components would primarily induce an increase in the Ca2+ cytosolic concentration, followed by membrane depolarization and activation of a plasma membrane NADPH oxidase. This latter effect, producing active oxygen species, might further influence plasma membrane permeability, leading to the massive release of electrolytes from the tissue. PMID- 9847113 TI - Biosynthesis of camalexin from tryptophan pathway intermediates in cell suspension cultures of Arabidopsis. AB - Camalexin (3-thiazol-2'-yl-indole) is the principal phytoalexin that accumulates in Arabidopsis after infection by fungi or bacteria. Camalexin accumulation was detectable in Arabidopsis cell-suspension cultures 3 to 5 h after inoculation with Cochliobolus carbonum (Race 1), and then increased rapidly from 7 to 24 h after inoculation. Levels of radioactivity incorporated into camalexin during a 1.5-h pulse labeling with [14C]anthranilate also increased with time after fungal inoculation. The levels of radioactive incorporation into camalexin increased rapidly between 7 and 18 h after inoculation, and then decreased along with camalexin accumulation. Relatively low levels of radioactivity from [14C]anthranilate incorporated into camalexin in the noninoculated controls. Autoradiographic analysis of the accumulation of chloroform-extractable metabolites labeled with [14C]anthranilate revealed a transient increase in the incorporation of radioactivity into indole in fungus-inoculated Arabidopsis cell cultures. The time-course measurement of radioactive incorporation into camalexin during a 1.5-h pulse labeling with [14C]indole was similar to that with [14C]anthranilate. These data suggest that indole destined for camalexin synthesis is produced by a separate enzymatic reaction that does not involve tryptophan synthase. PMID- 9847114 TI - High-molecular-weight FK506-binding proteins are components of heat-shock protein 90 heterocomplexes in wheat germ lysate. AB - In animal cell lysates the multiprotein heat-shock protein 90 (hsp90)-based chaperone complexes consist of hsp70, hsp40, and p60. These complexes act to convert steroid hormone receptors to their steroid-binding state by assembling them into heterocomplexes with hsp90, p23, and one of several immunophilins. Wheat germ lysate also contains a hsp90-based chaperone system that can assemble the glucocorticoid receptor into a functional heterocomplex with hsp90. However, only two components of the heterocomplex-assembly system, hsp90 and hsp70, have thus far been identified. Recently, purified mammalian p23 preadsorbed with JJ3 antibody-protein A-Sepharose pellets was used to isolate a mammalian p23-wheat hsp90 heterocomplex from wheat germ lysate (J.K. Owens-Grillo, L.F. Stancato, K. Hoffmann, W.B. Pratt, and P. Krishna [1996] Biochemistry 35: 15249-15255). This heterocomplex was found to contain an immunophilin(s) of the FK506-binding class, as judged by binding of the radiolabeled immunosuppressant drug [3H]FK506 to the immune pellets in a specific manner. In the present study we identified the immunophilin components of this heterocomplex as FKBP73 and FKBP77, the two recently described high-molecular-weight FKBPs of wheat. In addition, we present evidence that the two FKBPs bind hsp90 via tetratricopeptide repeat domains. Our results demonstrate that binding of immunophilins to hsp90 via tetratricopeptide repeat domains is a conserved protein interaction in plants. Conservation of this protein-to-protein interaction in both plant and animal cells suggests that it is important for the biological action of the high-molecular-weight immunophilins. PMID- 9847115 TI - Changes in hexokinase activity in echinochloa phyllopogon and echinochloa crus pavonis in response to abiotic stress AB - Hexokinase (HXK; EC 2.7.1.1) regulates carbohydrate entry into glycolysis and is known to be a sensor for sugar-responsive gene expression. The effect of abiotic stresses on HXK activity was determined in seedlings of the flood-tolerant plant Echinochloa phyllopogon (Stev.) Koss and the flood-intolerant plant Echinochloa crus-pavonis (H.B.K.) Schult grown aerobically for 5 d before being subjected to anaerobic, chilling, heat, or salt stress. HXK activity was stimulated in shoots of E. phyllopogon only by anaerobic stress. HXK activity was only transiently elevated in E. crus-pavonis shoots during anaerobiosis. In roots of both species, anoxia and chilling stimulated HXK activity. Thus, HXK is not a general stress protein but is specifically induced by anoxia and chilling in E. phyllopogon and E. crus-pavonis. In both species HXK exhibited an optimum pH between 8.5 and 9.0, but the range was extended to pH 7.0 in air-grown E. phyllopogon to 6.5 in N2 grown E. phyllopogon. At physiologically relevant pHs (6.8 and 7.3, N2 and O2 conditions, respectively), N2-grown seedlings retained greater HXK activity at the lower pH. The pH response suggests that in N2-grown seedlings HXK can function in a more acidic environment and that a specific isozyme may be important for regulating glycolytic activity during anaerobic metabolism in E. phyllopogon. PMID- 9847116 TI - The first step of gibberellin biosynthesis in pumpkin is catalyzed by at least two copalyl diphosphate synthases encoded by differentially regulated genes. AB - The first step in gibberellin biosynthesis is catalyzed by copalyl diphosphate synthase (CPS) and ent-kaurene synthase. We have cloned from pumpkin (Cucurbita maxima L.) two cDNAs, CmCPS1 and CmCPS2, that each encode a CPS. Both recombinant fusion CmCPS proteins were active in vitro. CPS are translocated into plastids and processed by cleavage of transit peptides. For CmCPS1 and CmCPS2, the putative transit peptides cannot exceed the first 99 and 107 amino acids, respectively, because longer N-terminal deletions abolished activity. Levels of both CmCPS transcripts were strictly regulated in an organ-specific and developmental manner. Both transcripts were almost undetectable in leaves and were abundant in petioles. CmCPS1 transcript levels were high in young cotyledons and low in roots. In contrast, CmCPS2 transcripts were undetectable in cotyledons but present at significant levels in roots. In hypocotyls, apices, and petioles, CmCPS1 transcript levels decreased with age much more rapidly than those of CmCPS2. We speculate that CmCPS1 expression is correlated with the early stages of organ development, whereas CmCPS2 expression is correlated with subsequent growth. In contrast, C. maxima ent-kaurene synthase transcripts were detected in every organ at almost constant levels. Thus, ent-kaurene biosynthesis may be regulated through control of CPS expression. PMID- 9847117 TI - Differential responses of abaxial and adaxial guard cells of broad bean to abscisic acid and calcium AB - Regulation by abscisic acid (ABA) and Ca2+ of broad bean (Vicia faba) abaxial and adaxial guard cell movements and inward K+ currents were compared. One millimolar Ca2+ in the bathing medium inhibited abaxial stomatal opening by 60% but only inhibited adaxial stomatal opening by 15%. The addition of 1 &mgr;M ABA in the bathing medium resulted in 80% inhibition of abaxial but only 45% inhibition of adaxial stomatal opening. Similarly, ABA and Ca2+ each stimulated greater abaxial stomatal closure than adaxial stomatal closure. Whole-cell patch-clamp results showed that the inward K+ currents of abaxial guard cells were inhibited by 60% ( 180 mV) in the presence of 1.5 &mgr;M Ca2+ in the cytoplasm, whereas the inward K+ currents of adaxial guard cells were not affected at all by the same treatment. Although 1 &mgr;M ABA in the cytoplasm inhibited the inward K+ currents to a similar extent for both abaxial and adaxial guard cells, the former were more sensitive to ABA applied externally. These results suggest that the abaxial stomata are more sensitive to Ca2+ and ABA than adaxial stomata in regard to stomatal opening and closing processes and that the regulation of the inward K+ currents by ABA may not proceed via a Ca2+-signaling pathway in adaxial guard cells. Therefore, there may be different pathways for ABA- and Ca2+-mediated signal transduction in abaxial and adaxial guard cells. PMID- 9847119 TI - Export of carbon from chloroplasts at night AB - Hexose export from chloroplasts at night has been inferred in previous studies of mutant and transgenic plants. We have tested whether hexose export is the normal route of carbon export from chloroplasts at night. We used nuclear magnetic resonance to distinguish glucose (Glc) made from hexose export and Glc made from triose export. Glc synthesized in vitro from fructose-6-phosphate in the presence of deuterium-labeled water had deuterium incorporated at C-2, whereas synthesis from triose phosphates caused C-2 through C-5 to become deuterated. In both tomato (Lycopersicon esculentum L. ) and bean (Phaseolus vulgaris L.), Glc from sucrose made at night in the presence of deuterium-enriched water was deuterated only in the C-2 position, indicating that >75% of carbon is exported as hexoses at night. In darkness the phosphate in the cytosol was 28 mM, whereas that in the chloroplasts was 5 mM, but hexose phosphates were 10-fold higher in the cytosol than in the chloroplasts. Therefore, hexose phosphates would not move out of chloroplasts without the input of energy. We conclude that most carbon leaves chloroplasts at night as Glc, maltose, or higher maltodextrins under normal conditions. PMID- 9847118 TI - Several thaumatin-like proteins bind to beta-1,3-glucans. AB - Pathogenesis-related proteins from intercellular fluid washings of stressed barley (Hordeum vulgare L.) leaves were analyzed to determine their binding to various water-insoluble polysaccharides. Three proteins (19, 16, and 15 kD) bound specifically to several water-insoluble beta-1,3-glucans. Binding of the barley proteins to pachyman occurred quickly at 22 degreesC at pH 5.0, even in the presence of 0.5 M NaCl, 0.2 M urea, and 1% (v/v) Triton X-100. Bound barley proteins were released by acidic treatments or by boiling in sodium dodecyl sulfate. Acid-released barley proteins could bind again specifically and singly to pachyman. Water-soluble laminarin and carboxymethyl-pachyman competed for the binding of the barley proteins to pachyman. The N-terminal sequence of the 19-kD barley beta-1,3-glucan-binding protein showed near identity to the barley seed protein BP-R and high homology to other thaumatin-like (TL) permatins. The 16-kD barley protein was also homologous to TL proteins, whereas the 15-kD barley protein N-terminal sequence was identical to the pathogenesis-related Hv-1 TL protein. Antifungal barley protein BP-R and corn (Zea mays) zeamatin were isolated by binding to pachyman. Two extracellular proteins from stressed pea (Pisum sativum L.) also bound to pachyman and were homologous to TL proteins. PMID- 9847120 TI - Direct measurement of calcium transport across chloroplast inner-envelope vesicles AB - The initial rate of Ca2+ movement across the inner-envelope membrane of pea (Pisum sativum L.) chloroplasts was directly measured by stopped-flow spectrofluorometry using membrane vesicles loaded with the Ca2+-sensitive fluorophore fura-2. Calibration of fura-2 fluorescence was achieved by combining a ratiometric method with Ca2+-selective minielectrodes to determine pCa values. The initial rate of Ca2+ influx in predominantly right-side-out inner-envelope membrane vesicles was greater than that in largely inside-out vesicles. Ca2+ movement was stimulated by an inwardly directed electrochemical proton gradient across the membrane vesicles, an effect that was diminished by the addition of valinomycin in the presence of K+. In addition, Ca2+ was shown to move across the membrane vesicles in the presence of a K+ diffusion potential gradient. The potential-stimulated rate of Ca2+ transport was slightly inhibited by diltiazem and greatly inhibited by ruthenium red. Other pharmacological agents such as LaCl3, verapamil, and nifedipine had little or no effect. These results indicate that Ca2+ transport across the chloroplast inner envelope can occur by a potential-stimulated uniport mechanism. PMID- 9847121 TI - Changes in salicylic acid and antioxidants during induced thermotolerance in mustard seedlings AB - Heat-acclimation or salicylic acid (SA) treatments were previously shown to induce thermotolerance in mustard (Sinapis alba L.) seedlings from 1.5 to 4 h after treatment. In the present study we investigated changes in endogenous SA and antioxidants in relation to induced thermotolerance. Thirty minutes into a 1 h heat-acclimation treatment glucosylated SA had increased 5.5-fold and then declined during the next 6 h. Increases in free SA were smaller (2-fold) but significant. Changes in antioxidants showed the following similarities after either heat-acclimation or SA treatment. The reduced-to-oxidized ascorbate ratio was 5-fold lower than the controls 1 h after treatment but recovered by 2 h. The glutathione pool became slightly more oxidized from 2 h after treatment. Glutathione reductase activity was more than 50% higher during the first 2 h. Activities of dehydroascorbate reductase and monodehydroascorbate reductase decreased by at least 25% during the first 2 h but were 20% to 60% higher than the control levels after 3 to 6 h. One hour after heat acclimation ascorbate peroxidase activity was increased by 30%. Young leaves appeared to be better protected by antioxidant enzymes following heat acclimation than the cotyledons or stem. Changes in endogenous SA and antioxidants may be involved in heat acclimation. PMID- 9847122 TI - Regulation of ribulose-1,5-bisphosphate Carboxylase/Oxygenase by carbamylation and 2-carboxyarabinitol 1-phosphate in tobacco: insights from studies of antisense plants containing reduced amounts of rubisco activase AB - The regulation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity by 2-carboxyarabinitol 1-phosphate (CA1P) was investigated using gas exchange analysis of antisense tobacco (Nicotiana tabacum) plants containing reduced levels of Rubisco activase. When an increase in light flux from darkness to 1200 &mgr;mol quanta m-2 s-1 was followed, the slow increase in CO2 assimilation by antisense leaves contained two phases: one represented the activation of the noncarbamylated form of Rubisco, which was described previously, and the other represented the activation of the CA1P-inhibited form of Rubisco. We present evidence supporting this conclusion, including the observation that this second phase, like CA1P, is only present following darkness or very low light flux. In addition, the second phase of CO2 assimilation was correlated with leaf CA1P content. When this novel phase was resolved from the CO2 assimilation trace, most of it was found to have kinetics similar to the activation of the noncarbamylated form of Rubisco. Additionally, kinetics of the novel phase indicated that the activation of the CA1P-inhibited form of Rubisco proceeds faster than the degradation of CA1P by CA1P phosphatase. These results may be significant with respect to current models of the regulation of Rubisco activity by Rubisco activase. PMID- 9847123 TI - Characterization of source- and sink-specific sucrose/H+ symporters from carrot. AB - To understand how sucrose (Suc) is transported from source leaves to developing tap roots of carrot (Daucus carota L.), we cloned two cDNAs (DcSUT1 and DcSUT2) for proteins with homologies to plant Suc/H+ symporters. The deduced polypeptide sequences are 52% identical and have 12 predicted membrane-spanning domains each. Transport activities were confirmed by expression of the clones in yeast cells. Both transporters had optimal activity below pH 5.0 and Michaelis constant values of 0.5 mM. Suc uptake was inhibited by protonophores, suggesting that Suc transport is linked to the proton electrochemical potential across the plasma membrane. DcSUT1 and DcSUT2 had markedly different expression patterns. Transcripts of DcSUT1 were found only in the green parts of plants, with highest levels in the lamina of source leaves, indicating that DcSUT1 is required for the loading of Suc into the phloem. In leaf lamina expression was diurnally regulated, suggesting that Suc export from the leaves is higher during the day than during the night. The mRNA of DcSUT2 was found mainly in sink organs, and no diurnal expression pattern was detected in the storage root. Here, expression was not restricted to the phloem but was much higher in storage parenchyma tissues of phloem and xylem. The close relationship of DcSUT2 with a Suc/H+ symporter from fava bean, which facilitates Suc uptake into the cotyledons of developing seeds, indicates that this carrot Suc transporter may be involved in loading Suc into storage parenchyma cells. PMID- 9847124 TI - Methyl jasmonate induces lauric acid omega-hydroxylase activity and accumulation of CYP94A1 transcripts but does not affect epoxide hydrolase activities in vicia sativa seedlings AB - Treatment of etiolated Vicia sativa seedlings by the plant hormone methyl jasmonate (MetJA) led to an increase of cytochrome P450 content. Seedlings that were treated for 48 h in a 1 mM solution of MetJA stimulated omega-hydroxylation of 12:0 (lauric acid) 14-fold compared with the control (153 versus 11 pmol min-1 mg-1 protein, respectively). Induction was dose dependent. The increase of activity (2.7-fold) was already detectable after 3 h of treatment. Activity increased as a function of time and reached a steady level after 24 h. Northern blot analysis revealed that the transcripts coding for CYP94A1, a fatty acid omega-hydroxylase, had already accumulated after 1 h of exposure to MetJA and was maximal between 3 and 6 h. Under the same conditions, a study of the enzymatic hydrolysis of 9,10-epoxystearic acid showed that both microsomal and soluble epoxide hydrolase activities were not affected by MetJA treatment. PMID- 9847125 TI - Potentiation of the oxidative burst and isoflavonoid phytoalexin accumulation by serine protease inhibitors AB - Treatment of soybean (Glycine max L. cv Williams 82) cell-suspension cultures with Pseudomonas syringae pv glycinea (Psg) harboring an avirulence gene (avrA) or with yeast elicitor resulted in an oxidative burst characterized by the accumulation of H2O2. This burst, and the resultant induction of glutathione S transferase transcripts, occurred more rapidly and was more prolonged if cells were simultaneously treated with serine protease inhibitors such as phenylmethylsulfonyl fluoride (PMSF) or diisopropylfluorophosphate. PMSF and diisopropylfluorophosphate potentiate a large oxidative burst in cells exposed to Psg harboring the avrC avirulence gene, which is not recognized by the soybean cultivar used in this study. The potentiated burst was inhibited by diphenylene iodonium, an inhibitor of NADPH oxidase, and by the protein kinase inhibitor K252a. PMSF treatment of elicited cells or cells exposed to Psg:avrA caused a large increase in the accumulation of the isoflavonoid phytoalexin glyceollin; however, this was not associated with increased levels of transcripts encoding key phytoalexin biosynthetic enzymes. Glyceollin accumulation was inhibited by diphenylene iodonium; however, the oxidative burst in cells treated with Psg:avrC and PMSF was not followed by phytoalexin accumulation. We conclude that active oxygen species from the oxidative burst are necessary but not sufficient for inducing isoflavonoid phytoalexin accumulation in soybean cells. PMID- 9847126 TI - Expression of beta-amylase from alfalfa taproots. AB - Alfalfa (Medicago sativa L.) roots contain large quantities of beta-amylase, but little is known about its role in vivo. We studied this by isolating a beta amylase cDNA and by examining signals that affect its expression. The beta amylase cDNA encoded a 55.95-kD polypeptide with a deduced amino acid sequence showing high similarity to other plant beta-amylases. Starch concentrations, beta amylase activities, and beta-amylase mRNA levels were measured in roots of alfalfa after defoliation, in suspension-cultured cells incubated in sucrose-rich or -deprived media, and in roots of cold-acclimated germ plasms. Starch levels, beta-amylase activities, and beta-amylase transcripts were reduced significantly in roots of defoliated plants and in sucrose-deprived cell cultures. beta-Amylase transcript was high in roots of intact plants but could not be detected 2 to 8 d after defoliation. beta-Amylase transcript levels increased in roots between September and October and then declined 10-fold in November and December after shoots were killed by frost. Alfalfa roots contain greater beta-amylase transcript levels compared with roots of sweetclover (Melilotus officinalis L.), red clover (Trifolium pratense L.), and birdsfoot trefoil (Lotus corniculatus L.). Southern analysis indicated that beta-amylase is present as a multigene family in alfalfa. Our results show no clear association between beta-amylase activity or transcript abundance and starch hydrolysis in alfalfa roots. The great abundance of beta-amylase and its unexpected patterns of gene expression and protein accumulation support our current belief that this protein serves a storage function in roots of this perennial species. PMID- 9847127 TI - pH-regulated leaf cell expansion in droughted plants is abscisic acid dependent AB - Elongation rates of barley (Hordeum vulgare L. cv Hanna) leaves decreased with decreasing soil water content, whereas the pH of xylem sap increased from 5.9 to 6.9 over 6 d as the soil dried. The reduction in leaf-elongation rate (LER) was correlated with the increase in sap pH. Artificial sap buffered to different pH values was fed via the subcrown internode to derooted seedlings. Although leaves elongated at in planta rates when fed artificial sap at a well-watered pH of 6.0, LER declined with increasing sap pH. This effect persisted in the light and in the dark. pH had no effect on the relative water content or the bulk abscisic acid (ABA) concentration of the growing zone of these leaves. LERs of the ABA deficient mutant Az34 were uniformly high over the pH range tested, whereas those of its isogenic wild-type cultivar Steptoe were reduced as the artificial sap pH was increased from 6.0 to 7.0. However, supplying a well-watered concentration of ABA (3 x 10(-8) M) in the artificial xylem sap restored the pH response of the Az34 mutant. The results suggest that increased xylem sap pH acts as a drought signal to reduce LER via an ABA-dependent mechanism. PMID- 9847129 TI - Carbohydrates in individual cells of epidermis, mesophyll, and bundle sheath in barley leaves with changed export or photosynthetic rate AB - Carbohydrate metabolism of barley (Hordeum vulgare) leaves induced to accumulate sucrose (Suc) and fructans was investigated at the single-cell level using single cell sampling and analysis. Cooling of the root and shoot apical meristem of barley plants led to the accumulation of Suc and fructan in leaf tissue. Suc and fructan accumulated in both mesophyll and parenchymatous bundle-sheath (PBS) cells because of the reduced export of sugars from leaves under cooling and to increased photosynthesis under high photon fluence rates. The general trends of Suc and fructan accumulation were similar for mesophyll and PBS cells. The fructan-to-Suc ratio was higher for PBS cells than for mesophyll cells, suggesting that the threshold Suc concentration needed for the initiation of fructan synthesis was lower for PBS cells. Epidermal cells contained very low concentrations of sugar throughout the cooling experiment. The difference in Suc concentration between control and treated plants was much less if compared at the single-cell level rather than the whole-tissue level, suggesting that the vascular tissue contains a significant proportion of total leaf Suc. We discuss the importance of analyzing complex tissues at the resolution of individual cells to assign molecular mechanisms to phenomena observed at the whole-plant level. PMID- 9847128 TI - Phytochrome regulates gibberellin biosynthesis during germination of photoblastic lettuce seeds. AB - Germination of lettuce (Lactuca sativa L.) seed is regulated by phytochrome. The requirement for red light is circumvented by the application of gibberellin (GA). We have previously shown that the endogenous content of GA1, the main bioactive GA in lettuce seeds, increases after red-light treatment. To clarify which step of GA1 synthesis is regulated by phytochrome, cDNAs encoding GA 20-oxidases (Ls20ox1 and Ls20ox2, for L. sativa GA 20-oxidase) and 3beta-hydroxylases (Ls3h1 and Ls3h2 for L. sativa GA 3beta-hydroxylase) were isolated from lettuce seeds by reverse-transcription polymerase chain reaction. Functional analysis of recombinant proteins expressed in Escherichia coli confirmed that the Ls20ox and Ls3h encode GA 20-oxidases and 3beta-hydroxylases, respectively. Northern-blot analysis showed that Ls3h1 expression was dramatically induced by red-light treatment within 2 h, and that this effect was canceled by a subsequent far-red light treatment. Ls3h2 mRNA was not detected in seeds that had been allowed to imbibe under any light conditions. Expression of the two Ls20ox genes was induced by initial imbibition alone in the dark. The level of Ls20ox2 mRNA decreased after the red-light treatment, whereas that of Ls20ox1 was unaffected by light. These results suggest that red light promotes GA1 synthesis in lettuce seeds by inducing Ls3h1 expression via phytochrome action. PMID- 9847130 TI - Molecular Biology Database List. AB - Molecular Biology Database List (MBDL) includes brief descriptions and pointers to Web sites for the various databases described in this issue as well as other Web sites presenting data sets relevant to molecular biology. This information is compiled into a list (http://www.oup.co.uk/nar/Volume_27/Issue_01/summary/ gkc105_gml.html) which includes links both to source Web sites and to on-line versions of articles describing the databases. PMID- 9847131 TI - DBcat: a catalog of biological databases. AB - The DBcat (http://www.infobiogen.fr/services/dbcat) is a comprehensive catalog of biological databases, maintained and curated on a daily basis at GIS Infobiogen. It contains more than 400 databases classified by application domains. The DBcat is a structured flat file library, that can be searched by means of an SRS server or a dedicated Web interface. The files are available for downloading from Infobiogen anonymous ftp server. PMID- 9847132 TI - GenBank. AB - The GenBank (Registered Trademark symbol) sequence database incorporates DNA sequences from all available public sources, primarily through the direct submission of sequence data from individual laboratories and from large-scale sequencing projects. Most submitters use the BankIt (Web) or Sequin programs to format and send sequence data. Data exchange with the EMBL Data Library and the DNA Data Bank of Japan helps ensure comprehensive worldwide coverage. GenBank data is accessible through NCBI's integrated retrieval system, Entrez, which integrates data from the major DNA and protein sequence databases along with taxonomy, genome and protein structure information. MEDLINE (Registered Trademark symbol) s from published articles describing the sequences are included as an additional source of biological annotation through the PubMed search system. Sequence similarity searching is offered through the BLAST series of database search programs. In addition to FTP, Email, and server/client versions of Entrez and BLAST, NCBI offers a wide range of World Wide Web retrieval and analysis services based on GenBank data. The GenBank database and related resources are freely accessible via the URL: http://www.ncbi.nlm.nih.gov PMID- 9847133 TI - The EMBL Nucleotide Sequence Database. AB - The EMBL Nucleotide Sequence Database (http://www.ebi.ac.uk/embl.html) constitutes Europe's primary nucleotide sequence resource. Main sources for DNA and RNA sequences are direct submissions from individual researchers, genome sequencing projects and patent applications. While automatic procedures allow incorporation of sequence data from large-scale genome sequencing centres and from the European Patent Office (EPO), the preferred submission tool for individual submitters is Webin (WWW). Through all stages, dataflow is monitored by EBI biologists communicating with the sequencing groups. In collaboration with DDBJ and GenBank the database is produced, maintained and distributed at the European Bioinformatics Institute (EBI). Database releases are produced quarterly and are distributed on CD-ROM. Network services allow access to the most up-to date data collection via Internet and World Wide Web interface. EBI's Sequence Retrieval System (SRS) is a Network Browser for Databanks in Molecular Biology, integrating and linking the main nucleotide and protein databases, plus many specialised databases. For sequence similarity searching a variety of tools (e.g. Blitz, Fasta, Blast etc) are available for external users to compare their own sequences against the most currently available data in the EMBL Nucleotide Sequence Database and SWISS-PROT. PMID- 9847134 TI - DNA Data Bank of Japan dealing with large-scale data submission. AB - The DNA Data Bank of Japan (DDBJ) (http//:www.ddbj.nig.ac.jp) has developed a software system for mass submissions to cope with a recent expansion of EST and genome data submissions. The system is composed of four parts, the WWW data submission, large-scale submission, submission management and storing. Using this system one can submit data on a large number of sequences or a very long sequence while checking the consistency between the annotation and sequence without much effort. DDBJ has received large scale data of Homo sapiens, Arabidopsis and Pyrococcus from Japanese researchers who made full use of the new submission system. PMID- 9847135 TI - KEGG: Kyoto Encyclopedia of Genes and Genomes. AB - Kyoto Encyclopedia of Genes and Genomes (KEGG) is a knowledge base for systematic analysis of gene functions in terms of the networks of genes and molecules. The major component of KEGG is the PATHWAY database that consists of graphical diagrams of biochemical pathways including most of the known metabolic pathways and some of the known regulatory pathways. The pathway information is also represented by the ortholog group tables summarizing orthologous and paralogous gene groups among different organisms. KEGG maintains the GENES database for the gene catalogs of all organisms with complete genomes and selected organisms with partial genomes, which are continuously re-annotated, as well as the LIGAND database for chemical compounds and enzymes. Each gene catalog is associated with the graphical genome map for chromosomal locations that is represented by Java applet. In addition to the data collection efforts, KEGG develops and provides various computational tools, such as for reconstructing biochemical pathways from the complete genome sequence and for predicting gene regulatory networks from the gene expression profiles. The KEGG databases are daily updated and made freely available (http://www.genome.ad.jp/kegg/). PMID- 9847136 TI - The Genome Sequence DataBase: towards an integrated functional genomics resource. AB - During 1998 the primary focus of the Genome Sequence DataBase (GSDB; http://www.ncgr.org/gsdb ) located at the National Center for Genome Resources (NCGR) has been to improve data quality, improve data collections, and provide new methods and tools to access and analyze data. Data quality has been improved by extensive curation of certain data fields necessary for maintaining data collections and for using certain tools. Data quality has also been increased by improvements to the suite of programs that import data from the International Nucleotide Sequence Database Collaboration (IC). The Sequence Tag Alignment and Consensus Knowledgebase (STACK), a database of human expressed gene sequences developed by the South African National Bioinformatics Institute (SANBI), became available within the last year, allowing public access to this valuable resource of expressed sequences. Data access was improved by the addition of the Sequence Viewer, a platform-independent graphical viewer for GSDB sequence data. This tool has also been integrated with other searching and data retrieval tools. A BLAST homology search service was also made available, allowing researchers to search all of the data, including the unique data, that are available from GSDB. These improvements are designed to make GSDB more accessible to users, extend the rich searching capability already present in GSDB, and to facilitate the transition to an integrated system containing many different types of biological data. PMID- 9847137 TI - The PIR-International Protein Sequence Database. AB - The Protein Information Resource (PIR; http://www-nbrf.georgetown. edu/pir/) supports research on molecular evolution, functional genomics, and computational biology by maintaining a comprehensive, non-redundant, well-organized and freely available protein sequence database. Since 1988 the database has been maintained collaboratively by PIR-International, an international association of data collection centers cooperating to develop this resource during a period of explosive growth in new sequence data and new computer technologies. The PIR Protein Sequence Database entries are classified into superfamilies, families and homology domains, for which sequence alignments are available. Full-scale family classification supports comparative genomics research, aids sequence annotation, assists database organization and improves database integrity. The PIR WWW server supports direct on-line sequence similarity searches, information retrieval, and knowledge discovery by providing the Protein Sequence Database and other supplementary databases. Sequence entries are extensively cross-referenced and hypertext-linked to major nucleic acid, literature, genome, structure, sequence alignment and family databases. The weekly release of the Protein Sequence Database can be accessed through the PIR Web site. The quarterly release of the database is freely available from our anonymous FTP server and is also available on CD-ROM with the accompanying ATLAS database search program. PMID- 9847138 TI - MIPS: a database for genomes and protein sequences. AB - The Munich Information Center for Protein Sequences (MIPS-GSF), Martinsried near Munich, Germany, develops and maintains genome oriented databases. It is commonplace that the amount of sequence data available increases rapidly, but not the capacity of qualified manual annotation at the sequence databases. Therefore, our strategy aims to cope with the data stream by the comprehensive application of analysis tools to sequences of complete genomes, the systematic classification of protein sequences and the active support of sequence analysis and functional genomics projects. This report describes the systematic and up-to-date analysis of genomes (PEDANT), a comprehensive database of the yeast genome (MYGD), a database reflecting the progress in sequencing the Arabidopsis thaliana genome (MATD), the database of assembled, annotated human EST clusters (MEST), and the collection of protein sequence data within the framework of the PIR-International Protein Sequence Database (described elsewhere in this volume). MIPS provides access through its WWW server (http://www.mips.biochem.mpg.de) to a spectrum of generic databases, including the above mentioned as well as a database of protein families (PROTFAM), the MITOP database, and the all-against-all FASTA database. PMID- 9847139 TI - The SWISS-PROT protein sequence data bank and its supplement TrEMBL in 1999. AB - SWISS-PROT is a curated protein sequence database which strives to provide a high level of annotation (such as the description of the function of a protein, its domain structure, post-translational modifications, variants, etc.), a minimal level of redundancy and high level of integration with other databases. Recent developments of the database include: cross-references to additional databases; a variety of new documentation files and improvements to TrEMBL, a computer annotated supplement to SWISS-PROT. TrEMBL consists of entries in SWISS-PROT-like format derived from the translation of all coding sequences (CDS) in the EMBL nucleotide sequence database, except the CDS already included in SWISS-PROT. The URLs for SWISS-PROT on the WWW are: http://www.expasy.ch/sprot and http://www. ebi.ac.uk/sprot PMID- 9847140 TI - Eco Cyc: encyclopedia of Escherichia coli genes and metabolism. AB - The EcoCyc database describes the genome and gene products of Escherichia coli, its metabolic and signal-transduction pathways, and its tRNAs. The database describes 4391 genes of E.coli, 695 enzymes encoded by a subset of these genes, 904 metabolic reactions that occur in E.coli, and the organization of these reactions into 129 metabolic pathways. The EcoCyc graphical user interface allows scientists to query and explore the EcoCyc database using visualization tools such as genomic-map browsers and automatic layouts of metabolic pathways. EcoCyc has many references to the primary literature, and is a (qualitative) computational model of E. coli metabolism. EcoCyc is available at URL http://ecocyc. PangeaSystems.com/ecocyc/ PMID- 9847141 TI - RegulonDB (version 2.0): a database on transcriptional regulation in Escherichia coli. AB - RegulonDB version 2.0, a database on transcriptional regulation and operon organization in Escherichia coli, is now available on the web at the following URL: http://www.cifn.unam. mx/Computational_Biology/regulondb/. In this paper we describe the main computational changes to the database, which include migrating the database to Sybase, providing graphical descriptions of the internal organization of operons and regulons, and direct links to MEDLINE references. The web interface offers searching either by mechanisms of regulation or by operon organization. The results of a search (operon organization, or site collection) are displayed as hypertext, and can also be displayed graphically. In terms of its contents, RegulonDB contains a large number of operons, as well as the absolute position in the completed genome sequence of sites, promoters, and individual genes of E.coli. PMID- 9847142 TI - RsGDB, the Rhodobacter sphaeroides Genome Database. AB - This report provides a summary of the sequencing project of the small chromosome (CII) of Rhodobacter sphaeroides 2.4.1(T),and introduces the first version of the genome database of this bacterium. The database organizes and describes diverse sets of biological information. The main role of the R.sphaeroides genome database (RsGDB) is to provide public access to the collected genomic information for R.sphaeroides via the World-Wide Web at http://utmmg.med.uth.tmc.edu/sphaeroides. The database allows the user access to hundreds of low redundancy R.sphaeroides sequences for further database searching, a summary of our current search results, and other allied information pertaining to this bacterium. PMID- 9847143 TI - The Enhanced Microbial Genomes Library. AB - Since the obtention of the complete sequence of Haemophilus influenzae Rd in 1995, the number of bacterial genomes entirely sequenced has regularly increased. A problem is that the quality of the annotations of these very large sequences is usually lower than those of the shorter entries encountered in the repository collections. Moreover, classical sequence database management systems have difficulties in handling entries of that size. In this context, we have decided to build the Enhanced Microbial Genomes Library (EMGLib) in which these two problems are alleviated. This library contains all the complete genomes from bacteria already sequenced and the yeast genome in GenBank format. The annotations are improved by the introduction of data on codon usage, gene orientation on the chromosome and gene families. It is possible to access EMGLib through two database systems set up on World Wide Web servers: the PBIL server at http://pbil.univ-lyon1.fr/emglib/emglib. html and the MICADO server at http://locus.jouy.inra.fr/micado PMID- 9847144 TI - Extension of CyanoBase. CyanoMutants: repository of mutant information on Synechocystis sp. strain PCC6803. AB - CyanoBase provides internet access to the complete genomic information of the cyanobacterium Synechocystis sp. strain PCC6803. CyanoBase contains annotations to each protein-coding gene, deduced from the entire nucleotide sequence of the genome, gene classification lists, keywords and similarity search engines. The present paper describes a recent extension of CyanoBase, named CyanoMutants. CyanoMutants is a repository database of mutant information on PCC6803. Each entry contains a dataset which describes a gene identifier, mutant information, and an address for correspondence. Two closely-linked databases, CyanoBase and CyanoMutants, connect information obtained from computational analysis to experimental analysis resulting in the clarification of the functions of hypothetical genes of the cyanobacterial genome. CyanoMutants can be accessed at http://www.kazusa.or. jp/cyano/mutants/ PMID- 9847145 TI - The Yeast Proteome Database (YPD): a model for the organization and presentation of genome-wide functional data. AB - The Yeast Proteome Database (YPD) is a model for the organization and presentation of comprehensive protein information. Based on the detailed curation of the scientific literature for the yeast Saccharomyces cerevisiae, YPD contains more than 50 000 annotations lines derived from the review of 8500 research publications. The information concerning each of the approximately 6100 yeast proteins is structured around a convenient one-page format, the Yeast Protein Report, with additional information provided as pop-up windows. Protein classification schema have been revised this year, defining each protein's cellular role, function and pathway, and adding a Functional to the Yeast Protein Report. These changes provide the user with a succinct summary of the protein's function and its place in the biology of the cell, and they enhance the power of YPD Search functions. Precalculated sequence alignments have been added, to provide a crossover point for comparative genomics. The first transcript profiling data has been integrated into the YPD Protein Reports, providing the framework for the presentation of genome-wide functional data. The Yeast Proteome Database can be accessed on the Web at http://www.proteome.com/YPDhome.html PMID- 9847146 TI - Using the Saccharomyces Genome Database (SGD) for analysis of protein similarities and structure. AB - The Saccharomyces Genome Database (SGD) collects and organizes information about the molecular biology and genetics of the yeast Saccharomyces cerevisiae. The latest protein structure and comparison tools available at SGD are presented here. With the completion of the yeast sequence and the Caenorhabditis elegans sequence soon to follow, comparison of proteins from complete eukaryotic proteomes will be an extremely powerful way to learn more about a particular protein's structure, its function, and its relationships with other proteins. SGD can be accessed through the World Wide Web at http://genome www.stanford.edu/Saccharomyces/ PMID- 9847147 TI - Unified display of Arabidopsis thaliana physical maps from AtDB, the A.thaliana database. AB - In the past several years, there has been a tremendous effort to construct physical maps and to sequence the genome of Arabidopsis thaliana. As a result, four of the five chromosomes are completely covered by overlapping clones except at the centromeric and nucleolus organizer regions (NOR). In addition, over 30% of the genome has been sequenced and completion is anticipated by the end of the year 2000. Despite these accomplishments, the physical maps are provided in many formats on laboratories' Web sites. These data are thus difficult to obtain in a coherent manner for researchers. To alleviate this problem, AtDB (Arabidopsis thaliana DataBase, URL: http://genome-www.stanford.edu/Arabidopsis/) has constructed a unified display of the physical maps where all publicly available physical-map data for all chromosomes are presented through the Web in a clickable, 'on-the-fly' graphic, created by CGI programs that directly consult our relational database. PMID- 9847149 TI - Grasping at molecular interactions and genetic networks in Drosophila melanogaster using FlyNets, an Internet database. AB - FlyNets (http://gifts.univ-mrs.fr/FlyNets/FlyNets_home_page.++ +html) is a WWW database describing molecular interactions (protein-DNA, protein-RNA and protein protein) in the fly Drosophila melanogaster. It is composed of two parts, as follows. (i) FlyNets-base is a specialized database which focuses on molecular interactions involved in Drosophila development. The information content of FlyNets-base is distributed among several specific lines arranged according to a GenBank-like format and grouped into five thematic zones to improve human readability. The FlyNets database achieves a high level of integration with other databases such as FlyBase, EMBL, GenBank and SWISS-PROT through numerous hyperlinks. (ii) FlyNets-list is a very simple and more general databank, the long-term goal of which is to report on any published molecular interaction occuring in the fly, giving direct web access to corresponding s in Medline and in FlyBase. In the context of genome projects, databases describing molecular interactions and genetic networks will provide a link at the functional level between the genome, the proteome and the transcriptome worlds of different organisms. Interaction databases therefore aim at describing the contents, structure, function and behaviour of what we herein define as the interactome world. PMID- 9847148 TI - The FlyBase database of the Drosophila Genome Projects and community literature. AB - The FlyBase Drosophila genetics database and the public interfaces of the Berkeley Drosophila Genome Project (BDGP) and European Drosophila Genome Project (EDGP) are in the process of integrating. At present, the data of these projects are available from independent, but hyperlinked, WWW sites (FlyBase URL, http://flybase. bio.indiana.edu/; BDGP URL, http://fruitfly.berkeley.edu/; EDGP URL, http://edgp.ebi.ac.uk/ ). Because of the considerable overlap of data classes between the contributions of the Drosophila genome projects and the Drosophila community, the new and enlarged FlyBase consortium views the implementation of a single integrated Drosophila genomics/genetics server as essential to the scientific community. This integration will occur in a stepwise fashion over the next 1-2 years. In this report, the salient features of the current databases and how to interrogate and navigate the extensive data sets are discussed. PMID- 9847150 TI - The Mouse Genome Database (MGD): genetic and genomic information about the laboratory mouse. The Mouse Genome Database Group. AB - The Mouse Genome Database (MGD) focuses on the integration of mapping, homology, polymorphism and molecular data about the laboratory mouse. Detailed descriptions of genes including their chromosomal location, gene function, disease associations, mutant phenotypes, molecular polymorphisms and links to representative sequences including ESTs are integrated within MGD. The association of information from experiment to gene to genome requires careful coordination and implementation of standardized vocabularies, unique nomenclature constructions, and detailed information derived from multiple sources. This information is linked to other public databases that focus on additional information such as expression patterns, sequences, bibliographic details and large mapping panel data. Scientists participate in the curation of MGD data by generating the Chromosome Committee Reports, consulting on gene family nomenclature revisions, and providing descriptions of mouse strain characteristics and of new mutant phenotypes. MGD is accessible at http://www.informatics.jax.org PMID- 9847151 TI - Electronic access to mouse tumor data: the Mouse Tumor Biology Database (MTB) project. AB - The Mouse Tumor Biology (MTB) Database supports the use of the mouse as a model system of hereditary and induced cancers by providing electronic access to: (i) tumor names and classifications, (ii) tumor incidence and latency data in different strains of mice, (iii) tumor pathology reports and images, (iv) information on genetic factors associated with tumors and tumor development, and (v) references (published and unpublished data). This resource has been designed to aid researchers in such areas as choosing experimental models, reviewing patterns of mutations in specific cancers, and identifying genes that are commonly mutated across a spectrum of cancers. MTB also provides hypertext links to related on-line resources and databases. MTB is accessible via the World Wide Web at http://tumor.informatics.jax.org. User support is available for MTB by Email at mgi-help@informatics.jax.org PMID- 9847152 TI - GXD: a gene expression database for the laboratory mouse. The Gene Expression Database Group. AB - The Gene Expression Database (GXD) is a community resource that stores and integrates expression information for the laboratory mouse, with a particular emphasis on mouse development, and makes these data freely available in formats appropriate for comprehensive analysis. GXD is implemented as a relational database and integrated with the Mouse Genome Database (MGD) to enable global analysis of genotype, expression and phenotype information. Interconnections with sequence databases and with databases from other species further extend GXD's utility for the analysis of gene expression data. GXD is available through the Mouse Genome Informatics Web Site at http://www.informatics.jax.org/ PMID- 9847153 TI - Virgil database for rich links (1999 update). AB - With so many databases available for research in the Human Genome Project, it is crucial to efficiently relate information from different resources. For that purpose, we maintain Virgil, a database of rich links for data browsing, data analysis and database interconnection. Virgil current version contains more than 40 000 rich links from five major databases: SWISS-PROT, GenBank, PDB, GDB and OMIM. Materials described in this paper are available from http://www.infobiogen.fr/services/virgil/ PMID- 9847155 TI - The HuGeMap Database: interconnection and visualization of human genome maps. AB - The HuGeMap database stores the major genetic and physical maps of the human genome. HuGeMap is accessible on the Web at http://www. infobiogen.fr/services/Hugemap and through a CORBA server. A standard genome map data format for the interconnection of genome map databases was defined in collaboration with the EBI. The HuGeMap CORBA server provides this interconnection using the interface definition language IDL. Two graphical user interfaces were developed for the visualization of the HuGeMap data: ZoomMap (http://www.infobiogen.fr/services/zomit/Zoom Map.html) for navigation by zooming and data transformation via magic lenses, and MappetShow (http://www.infobiogen.fr/services/Mappet) for visualizing and comparing maps. PMID- 9847154 TI - The radiation hybrid database. AB - Since July 1995, the European Bioinformatics Institute (EBI) has maintained the Radiation Hybrid database (RHdb; http://www.ebi.ac. uk/RHdb ), a public database for radiation hybrid data. Radiation hybrid mapping is an important technique for determining high resolution maps. Recently, CORBA access has been added to RHdb. The EBI is an Outstation of the European Molecular Biology Laboratory (EMBL). PMID- 9847156 TI - IXDB, an X chromosome integrated database (update). AB - Chromosome specific databases are an important research tool as they integrate data from different directions, such as genetic and physical mapping data, expression data, sequences etc. They supplement the genome-wide repositories in molecular biology, such as GenBank, Swiss-Prot or OMIM, which usually concentrate on one type of information. The Integrated X Chromosome Database (IXDB, http://ixdb.mpimg-berlin-dahlem.mpg.de/) is a repository for physical mapping data of the human X chromosome and aims at providing a global view of genomic data at a chromosomal level. We present here an update of IXDB which includes schema extensions for storing submaps and sequence information, additional links to external databases, and the integration of an increasing number of physical and transcript mapping data. The gene data was completely updated according to the approved gene symbols of the HUGO Nomenclature Committee. IXDB receives over 1000 queries per month, an indication that its content is valuable to researchers seeking mapping data of the human X chromosome. PMID- 9847158 TI - Update of AMmtDB: a database of multi-aligned metazoa mitochondrial DNA sequences. AB - The present paper describes AMmtDB, a database collecting the multi-aligned sequences of vertebrate mitochondrial genes coding for proteins and tRNAs, as well as the multiple alignment of the mammalian mtDNA main regulatory region (D loop) sequences. The genes coding for proteins are multi-aligned based on the translated sequences and both the nucleotide and amino acid multi-alignments are provided. As far as the genes coding for tRNAs are concerned, the multi alignments based on the primary and the secondary structures are both provided; for the mammalian D-loop multi-alignments we report the conserved regions of the entire D-loop (CSB1, CSB2, CSB3, the central region, ETAS1 and ETAS2) as defined by Sbisa et al. [ Gene (1997), 205, 125-140). A flatfile format for AMmtDB has been designed allowing its implementation in SRS (http://bio www.ba.cnr.it:8000/BioWWW/#AMMTDB ). Data selected through SRS can be managed using GeneDoc or other programs for the management of multi-aligned data depending on the user's operative system. The multiple alignments have been produced with CLUSTALV and PILEUP programs and then carefully optimized manually. PMID- 9847157 TI - MitBASE: a comprehensive and integrated mitochondrial DNA database. AB - MitBASE is an integrated and comprehensive database of mitochondrial DNA data which collects all available information from different organisms and from intraspecie variants and mutants. Research institutions from different countries are involved, each in charge of developing, collecting and annotating data for the organisms they are specialised in. The design of the actual structure of the database and its implementation in a user-friendly format are the care of the European Bioinformatics Institute. The database can be accessed on the Web at the following address: http://www.ebi.ac. uk/htbin/Mitbase/mitbase.pl. The impact of this project is intended for both basic and applied research. The study of mitochondrial genetic diseases and mitochondrial DNA intraspecie diversity are key topics in several biotechnological fields. The database has been funded within the EU Biotechnology programme. PMID- 9847159 TI - HvrBase: compilation of mtDNA control region sequences from primates. AB - HvrBase is a compilation of human and ape mtDNA control region sequences. Sequences and related information on individuals, such as from where the sequences were obtained, is stored in three ASCII files as described previously. Moreover, the collection is also available as Mac/PC database application with a graphical user interface. It can be accessed through the WWW at URL http://www.eva.mpg.de/hvrbase. The current collection comprises 5846 human sequences from hypervariable region I (HVRI) and 2302 human sequences from hypervariable region II (HVRII). From apes, 295 HVRI sequences and 13 HVRII sequences are available. PMID- 9847160 TI - Update of the Human MitBASE database. AB - Human MitBASE is a database collecting human mtDNA variants. This database is part of a greater mitochondrial genome database (MitBASE) funded within the EU Biotech Program. The present paper reports the recent improvements in data structure, data quality and data quantity. As far as the database structure is concerned it is now fully designed and implemented. Based on the previously described structure some changes have been made to optimise both data input and data quality. Cross-references with other bio-databases (EMBL, OMIM, MEDLINE) have been implemented. Human MitBASE data can be queried with the MitBASE Simple Query System (http://www.ebi.ac.uk/htbin/Mitbase/mit base.pl) and with SRS at the EBI under the 'Mutation' section (http://srs.ebi.ac.uk/srs5/). At present the HumanMitBASE node contains approximately 5000 variants related to studies investigating population polymorphisms and pathologies. PMID- 9847161 TI - MitBASE pilot: a database on nuclear genes involved in mitochondrial biogenesis and its regulation in Saccharomyces cerevisiae. AB - In the framework of the EU BIOTECH PROGRAM and within the 'MITBASE: a comprehensive and integrated database on mtDNA' project, we have prepared a pilot database (MitBASE Pilot) on nuclear genes involved in mitochondrial biogenesis and its regulation in Saccharomyces cerevisiae. MitBASE Pilot includes nuclear genes encoding mitochondrial proteins as well as nuclear genes encoding products which are localised in other sub-cellular compartments but nevertheless interact with mitochondrial functions. Genes have been classified on the basis of the mitochondrial process in which they participate and the mitochondrial phenotype of the gene knockout. The structure of the MitBASE Pilot database has been conceived for a flexible organisation of the information. An intuitive visual query system has been developed which allows users to select information in different combinations, both in the query and the output format, according to their needs. MitBASE Pilot is a relational database, is maintained at the EMBL European Bioinformatics Institute (EBI) and is available at the World Wide Web site http://www3.ebi.ac. uk/Research/Mitbase/mitbiog.pl PMID- 9847162 TI - Vertebrate MitBASE: a specialised database on vertebrate mitochondrial DNA sequences. AB - Vertebrate MitBASE is a specialized database where all the vertebrate mitochondrial DNA entries from primary databases are collected, revised and integrated with new information emerging from the literature. Variant sequences are also analyzed, aligned and linked to reference sequences. Data related to the same species and fragment can be viewed over the WWW. The database has a flexible interface and a retrieval system to help non-expert users and contains information not currently available in the primary databases. Vertebrate MitBASE is now available through the MitBASE home page at URL: http://www.ebi.ac.uk/htbin/Mitbase/mitb ase.pl. This work is part of a larger project, MitBASE which is a network of databases covering the full panorama of knowledge on mitochondrial DNA from protists to human sequences. PMID- 9847163 TI - MITOP: database for mitochondria-related proteins, genes and diseases. AB - The MITOP database http://websvr.mips.biochem.mpg. de/proj/medgen/mitop/ consolidates information on both nuclear- and mitochondrial-encoded genes and their proteins. The five species files- Saccharomyces cerevisiae, Mus musculus, Caenorhabditis elegans, Neurospora crassa and Homo sapiens -include annotated data derived from a variety of online resources and the literature. A wide spectrum of search facilities is given in the interelated sections 'Gene catalogues', 'Protein catalogues', 'Homologies', 'Pathways and metabolism', and 'Human disease catalogue' including extensive references and hyperlinks for each entry. Precomputed FASTA searches using all the MITOP yeast protein entries and a list of the best EST hits with graphical cluster alignments related to the yeast reference sequence are presented. The MITOP orthologue tables with cross-listing to all the protein entries for each species in the database facilitate investigations into interspecies homology. A program (MITOPROT) is available to identify mitochondrial targeting sequences and graphical depictions of several important mitochondrial processes are included. The 'Human disease catalogue' lists a total of 101 disorders related to mitochondrial protein abnormalities, sorted by clinical criteria and age of onset. PMID- 9847164 TI - PLMItRNA, a database for higher plant mitochondrial tRNAs and tRNA genes. AB - The PLMItRNA database contains information and multialignments of tRNA genes and molecules detected in higher plant mitochondria. It has been developed from a previous compilation of higher plant mitochondrial tRNA genes [Sagliano,A., Volpicella,M., Gallerani,R. and Ceci,L.R. (1998) Nucleic Acids Res., 26, 154-155] and implemented with data and sequences of tRNA molecules retrieved from the literature. The current version of the database reports information on 171 genes and 16 tRNA molecules from 24 plants. PLMItRNA is accessible via WWW at http://bio-www.ba.cnr.it:8000/srs/ PMID- 9847165 TI - 5S Ribosomal RNA Data Bank. AB - This paper presents the updated version of the data base of ribosomal 5S ribonucleic acids (5S rRNA) and their genes (5S rDNA). This edition of the data bank contains 1889 primary structures of 5S rRNA and 5S rDNA. These include 60 archaebacterial, 439 eubacterial, 63 plastid, 9 mitochondrial and 1318 eukaryotic sequences. The nucleotide sequences of 5S rRNAs or 5S rDNAs are divided according to the taxonomic position of organisms. The sequences stored in the database can be viewed and retrieved using the taxonomic browser at the URL: http://rose.man.poznan.pl/5SData/5SRNA.html++ + PMID- 9847166 TI - A comprehensive database for the small nucleolar RNAs from Saccharomyces cerevisiae. AB - Small nucleolar RNAs (snoRNAs) are involved in cleavage of rRNA, modification of rRNA nucleotides and, perhaps, other aspects of ribosome biogenesis in eukaryotic cells. Scores of snoRNAs have been discovered in recent years from various eukaryotes, and the total number is predicted to be up to 200 different snoRNA species per individual organism. We have created a comprehensive database for snoRNAs from the yeast Saccharomyces cerevisiae which allows easy access to detailed information about each species known (almost 70 snoRNAs are featured). The database consists of three major parts: (i) a utilities section; (ii) a master table; and (iii) a collection of tables for the individual snoRNAs. The utilities section provides an introduction to the database. The master table lists all known S. cerevisiae snoRNAs and their major properties. Information in the individual tables includes: alternate names, size, family classification, genomic organization, sequences (with major features identified), GenBank accession numbers, occurrence of homologues, gene disruption phenotypes, functional properties and associated RNAs and proteins. All information is accompanied with appropriate literature references. The database is available on the World Wide Web (http://www.bio.umass. edu/biochem/rna sequence/Yeast_snoRNA_Database/snoRNA_ DataBase.html), and should be useful for a wide range of snoRNA studies. PMID- 9847167 TI - The tmRNA website. AB - tmRNA (10Sa RNA) has a central role in trans -translation, in which a peptide tag encoded in tmRNA is added to the abnormally short protein product of a broken mRNA, as a signal for proteolysis of the entire tagged protein. The tmRNA website was established in 1997 as a resource for phylogenetic considerations of tmRNA structure and function. Since then, three partial tmRNA sequences have been completed, and sequences from 13 more species have been identified. Forty-six species from 10 bacterial phyla and chloroplasts are now represented in the database. Provisional sequence alignments and predicted proteolysis tag sequences are provided, as well as a literature review and a guide to searching for new tmRNA sequences. The tmRNA website is accessible via WWW at a new URL: http://sunflower.bio.indiana.edu/kwilliam/tmRNA /home.html PMID- 9847168 TI - The tmRNA database (tmRDB). AB - As of September, 1998, a total of 43 sequences are contained within the tmRNA database (tmRDB). The tmRNA sequences are arranged alphabetically and ordered phylogenetically. The alignment of the tmRNAs emphasizes the basepairs that are supported by comparative sequence analysis and establishes minimal secondary structures for the known tmRNAs. A corresponding alignment of the predicted tmRNA encoded tag peptides is presented. The tmRDB also offers a small number of RNA secondary structure diagrams and PDB-formatted three-dimensional models generated with the program ERNA-3D. The data are available freely at the URL http://psyche.uthct.edu/dbs/tmRDB/tmRDB.++ +html PMID- 9847169 TI - The guide RNA database (3.0). AB - The RNA editing process within the mitochondria of kinetoplastid organisms is controlled by small, trans -acting RNA molecules referred to as guide RNAs. The guide RNA database is a compilation of published guide RNA sequences, currently containing 254 entries from 11 different organisms. Additional information includes RNA secondary and tertiary structure models, information on the gene localisation, literature citations and other relevant facts. The database can be accessed through the World Wide Web (WWW) at http://www.biochem.mpg.de/ goeringe/ PMID- 9847170 TI - The Signal Recognition Particle Database (SRPDB). AB - The signal recognition particle database (SRPDB) is located at the University of Texas Health Science Center at Tyler and includes tabulations of SRP RNA, SRP protein and SRP receptor sequences. The sequences are annotated with links to the primary databases. They are ordered alphabetically or phylogenetically and are available in aligned form. As of September, 1998, there were 108 SRP RNA sequences, 83 SRP protein sequences and 28 sequences of the SRP receptor alpha subunit and its homologues. In addition, the SRPDB provides search motifs consisting of conserved amino acid and nucleotide residues, and a limited number of SRP RNA secondary structure diagrams and 3-D models. The data are available freely at the URL http://psyche.uthct.edu/dbs/SRPDB/SRPDB.++ +html PMID- 9847171 TI - A new version of the RDP (Ribosomal Database Project). AB - The Ribosomal Database Project (RDP-II), previously described by Maidak et al. [ Nucleic Acids Res. (1997), 25, 109-111], is now hosted by the Center for Microbial Ecology at Michigan State University. RDP-II is a curated database that offers ribosomal RNA (rRNA) nucleotide sequence data in aligned and unaligned forms, analysis services, and associated computer programs. During the past two years, data alignments have been updated and now include >9700 small subunit rRNA sequences. The recent development of an ObjectStore database will provide more rapid updating of data, better data accuracy and increased user access. RDP-II includes phylogenetically ordered alignments of rRNA sequences, derived phylogenetic trees, rRNA secondary structure diagrams, and various software programs for handling, analyzing and displaying alignments and trees. The data are available via anonymous ftp (ftp.cme.msu. edu) and WWW (http://www.cme.msu.edu/RDP). The WWW server provides ribosomal probe checking, approximate phylogenetic placement of user-submitted sequences, screening for possible chimeric rRNA sequences, automated alignment, and a suggested placement of an unknown sequence on an existing phylogenetic tree. Additional utilities also exist at RDP-II, including distance matrix, T-RFLP, and a Java-based viewer of the phylogenetic trees that can be used to create subtrees. PMID- 9847172 TI - Database on the structure of large subunit ribosomal RNA. AB - The Antwerp database on large subunit ribosomal RNA now contains 607 complete or nearly complete aligned sequences. The alignment incorporates secondary structure information for each sequence. Other information about the sequences, such as literature references, accession numbers and taxonomic information is also available. Information from the database can be downloaded or searched on the rRNA WWW Server at URL http://rrna.uia.ac.be/ PMID- 9847173 TI - Database on the structure of small subunit ribosomal RNA. AB - Over 11 500 complete or nearly complete sequences are now available from the Antwerp database on small subunit ribosomal RNA. All these sequences are aligned with one another on the basis of the adopted secondary structure model, which is corroborated by the observation of compensating substitutions in the alignment. Literature references, accession numbers and taxonomic information are also compiled. The database can be consulted via the World Wide Web at URL http://rrna.uia.ac.be/ssu/ PMID- 9847174 TI - The Database of Ribosomal Cross-links: an update. AB - The Database of Ribosomal Cross-links (DRC) was created in 1997. Here we describe new data incorporated into this database and several new features of the DRC. The DRC is freely available via World Wide Web at http://visitweb.com/database/ or http://www. mpimg-berlin-dahlem.mpg.de/ approximately ag_ribo/ag_brimacombe/drc/ PMID- 9847175 TI - The viroid and viroid-like RNA database. AB - This is an online database to facilitate research on viroid, viroid-like RNAs and human hepatitis delta virus (vHDV) by presenting a large number of sequences and related data in a comprehensive and user-friendly format (e.g. position of their self-catalytic domains, open reading frame of the vHDV, prediction of the most stable secondary structures, etc.). Most of these RNA species share a common proposed replication pattern known as a DNA-independent rolling circle mechanism. Together, these species form the 'brotherhood' of the smallest known auto replicable RNAs. This online database is available on the World Wide Web at http://www.callisto.si.usherb.ca/jpperra PMID- 9847176 TI - UTRdb: a specialized database of 5' and 3' untranslated regions of eukaryotic mRNAs. AB - The 5' and 3' untranslated regions of eukaryotic mRNAs may play a crucial role in the regulation of gene expression controlling mRNA localization, stability and translational efficiency. For this reason we developed UTRdb (http://bigarea.area.ba.cnr.it:8000/BioWWW/#U TRdb), a specialized database of 5' and 3' untranslated sequences of eukaryotic mRNAs cleaned from redundancy. UTRdb entries are enriched with specialized information not present in the primary databases including the presence of nucleotide sequence patterns already demonstrated by experimental analysis to have some functional role. All these patterns have been collected in the UTRsite database so that it is possible to search any input sequence for the presence of annotated functional motifs. Furthermore, UTRdb entries have been annotated for the presence of repetitive elements. PMID- 9847177 TI - Collection of mRNA-like non-coding RNAs. AB - In last few years much data has accumulated which shows that in different cells various RNA transcripts are synthesized. They lack protein coding capacity and do not produce mature protein. It seems that they work mainly or exclusively on the RNA level. Their function and mechanism of action is poorly understood. In this paper we have collected all known RNA transcript and prepared a database for further structural and functional studies. This is the first collection of the nucleotide sequences of RNAs of this kind. The data can be accessed via WWW at: http://www.man.poznan.pl/5SData/ncRNA/inde x.html PMID- 9847178 TI - The RNA Modification Database: 1999 update. AB - The RNA Modification Database (http://medlib.med.utah.edu/RNAmods/) provides a comprehensive listing of naturally modified nucleosides in RNA. Each file includes: chemical structure; common name and symbol; type(s) of RNA in which found and corresponding phylogenetic distribution; Chemical s registry number and index name; and initial literature citations for structure characterization and chemical synthesis. New features include capability to search database files by name or substructural features, modifications in tmRNA, and links to related data and sites. PMID- 9847179 TI - The RESID Database of protein structure modifications. AB - Because the number of post-translational modifications requiring standardized annotation in the PIR-International Protein Sequence Database was large and steadily increasing, a database of protein structure modifications was constructed in 1993 to assist in producing appropriate feature annotations for covalent binding sites, modified sites and cross-links. In 1995 RESID was publicly released as a PIR-International text database distributed on CD-ROM and accessible through the ATLAS program. In 1998 it was made available on the PIR Web site at http://www-nbrf.georgetown.edu/pir/searchdb++ +.html . The RESID Database includes such information as: systematic and frequently observed alternate names; Chemical s Service registry numbers; atomic formulas and weights; enzyme activities; indicators forN-terminal, C-terminal or peptide chain cross-link modifications; keywords; and literature citations with database cross references. The RESID Database can be used to predict atomic masses for peptides, and is being enhanced to provide molecular structures for graphical presentation on the PIR Web site using widely available molecular viewing programs. PMID- 9847180 TI - EpoDB: a prototype database for the analysis of genes expressed during vertebrate erythropoiesis. AB - EpoDB is a database of genes expressed in vertebrate red blood cells. It is also a prototype for the creation of cell and tissue-specific databases from multiple external sources. The information in EpoDB obtained from GenBank, SWISS-PROT, Transfac, TRRD and GERD is curated to provide high quality data for sequence analysis aimed at understanding gene regulation during erythropoiesis. New protocols have been developed for data integration and updating entries. Using a BLAST-based algorithm, we have grouped GenBank entries representing the same gene together. This sequence similarity protocol was also used to identify new entries to be included in EpoDB. We have recently implemented our database in Sybase (relational tables) in addition to SICStus Prolog to provide us with greater flexibility in asking complex queries that utilize information from multiple sources. New additions to the public web site (http://www.cbil.upenn.edu/epodb) for accessing EpoDB are the ability to retrieve groups of entries representing different variants of the same gene and to retrieve gene expression data. The BLAST query has been enhanced by incorporating BLASTView, an interactive and graphical display of BLAST results. We have also enhanced the queries for retrieving sequence from specified genes by the addition of MEME, a motif discovery tool, to the integrated analysis tools which include CLUSTALW and TESS. PMID- 9847182 TI - IMGT, the international ImMunoGeneTics database. AB - IMGT, the international ImMunoGeneTics database (http://imgt.cnusc. fr:8104), is a high-quality integrated database specialising in Immunoglobulins (Ig), T cell Receptors (TcR) and Major Histocompatibility Complex (MHC) molecules of all vertebrate species, created in 1989 by Marie-Paule Lefranc, Universite Montpellier II, CNRS, Montpellier, France (lefranc@ligm.igh.cnrs.fr). IMGT comprises three databases: LIGM-DB, a comprehensive database of Ig and TcR, MHC/HLA-DB, and PRIMER-DB (the last two in development); a tool, IMGT/DNAPLOT, developed for sequence analysis and alignments; and expertised data based on the IMGT scientific chart, the IMGT repertoire. By its high quality and its easy data distribution, IMGT has important implications in medical research (repertoire in autoimmune diseases, AIDS, leukemias, lymphomas), therapeutic approaches (antibody engineering), genome diversity and genome evolution studies. IMGT is freely available at http://imgt.cnusc. fr:8104 PMID- 9847181 TI - PEDB: the Prostate Expression Database. AB - The Prostate Expression Database (PEDB) is a curated relational database and suite of analysis tools designed for the study of prostate gene expression in normal and disease states. Expressed Sequence Tags (ESTs) and full-length cDNA sequences derived from more than 40 human prostate cDNA libraries are maintained and represent a wide spectrum of normal and pathological conditions. Detailed library information including tissue source, library construction methods, sequence diversity and abundance are available in a library archive. Prostate ESTs are assembled into distinct species groups using the multiple alignment program CAP2 and are annotated with information from the GenBank, dbEST and Unigene public sequence databases. Annotated sequences in PEDB are searched using the BLAST algorithm. The differential expression of each EST species can be viewed across all libraries using a Virtual Expression Analysis Tool (VEAT), a graphical user interface written in Java for intra- and inter-library species comparisons. PEDB may be accessed via the World Wide Web at http://www.mbt.washington.edu/PEDB/ PMID- 9847183 TI - Current status of the Asthma and Allergy Database. AB - The database provides an online resource for access to data on the genetics of asthma and allergy. This report describes the present status of the site. Currently, a detailed description of 88 linkage studies (7164 linkage positions) and 72 mutation studies are available. The results can be accessed in table form or graphically. The database also contains mouse asthma studies and human homology relationships, gene expression studies and links to relevant patents. Technical details about the server architecture, database installation, database construction, database structure and the user interface are explained elsewhere [Wjst and Immervoll (1998) Bioinformatics, 14, 827-828]. The URL is http://cooke.gsf.de PMID- 9847184 TI - The PROSITE database, its status in 1999. AB - The PROSITE database (http://www.expasy.ch/sprot/prosite.htm l) consists of biologically significant patterns and profiles formulated in such a way that with appropriate computational tools it can help to determine to which known family of protein (if any) a new sequence belongs, or which known domain(s) it contains. PMID- 9847185 TI - PRINTS prepares for the new millennium. AB - PRINTS is a diagnostic collection of protein fingerprints. Fingerprints exploit groups of motifs to build characteristic family signatures, offering improved diagnostic reliability over single-motif approaches by virtue of the mutual context provided by motif neighbours. Around 1000 fingerprints have now been created and stored in PRINTS. The September 1998 release (version 20.0), encodes approximately 5700 motifs, covering a range of globular and membrane proteins, modular polypeptides and so on. The database is accessible via the DbBrowser Web Server at http://www.biochem.ucl.ac.uk/bsm/dbbrowser /. In addition to supporting its continued growth, recent enhancements to the resource include a BLAST server, and more efficient fingerprint search software, with improved statistics for estimating the reliability of retrieved matches. Current efforts are focused on the design of more automated methods for database maintenance; implementation of an object-relational schema for efficient data management; and integration with PROSITE, profiles, Pfam and ProDom, as part of the international InterPro project, which aims to unify protein pattern databases and offer improved tools for genome analysis. PMID- 9847186 TI - New features of the Blocks Database servers. AB - Blocks are ungapped multiple sequence alignments representing conserved protein regions, and the Blocks Database consists of blocks from documented protein families. World Wide Web (http://www. blocks.fhcrc.org) and Email (blocks@blocks.fhcrc.org) servers provide tools for homology searching and for analyzing protein family relationships. New enhancements include a multiple alignment processor that extends the use of these tools to imported multiple alignments of families not present in the database and a PCR primer designer that implements a new strategy for gene isolation. PMID- 9847187 TI - SMART: identification and annotation of domains from signalling and extracellular protein sequences. AB - SMART is a simple modular architecture research tool and database that provides domain identification and annotation on the WWW (http://coot.embl heidelberg.de/SMART). The tool compares query sequences with its databases of domain sequences and multiple alignments whilst concurrently identifying compositionally biased regions such as signal peptide, transmembrane and coiled coil segments. Annotated and unannotated regions of the sequence can be used as queries in searches of sequence databases. The SMART alignment collection represents more than 250 signalling and extracellular domains. Each alignment is curated to assign appropriate domain boundaries and to ensure its quality. In addition, each domain is annotated extensively with respect to cellular localisation, species distribution, functional class, tertiary structure and functionally important residues. PMID- 9847188 TI - PROMISE: a database of bioinorganic motifs. AB - The PROMISE (prosthetic centres andmetalions in protein activesites) database aims to present comprehensive sequence, structural, functional and bibliographic information on metalloproteins and other complex proteins, with an emphasis on active site structure and function. The database is available on the WorldWide Web at http://bioinf.leeds.ac.uk/promise/ PMID- 9847189 TI - PhosphoBase, a database of phosphorylation sites: release 2.0. AB - PhosphoBase contains information about phosphorylated residues in proteins and data about peptide phosphorylation by a variety of protein kinases. The data are collected from literature and compiled into a common format. The current release of PhosphoBase (October 1998, version 2.0) comprises 414 phosphoprotein entries covering 1052 phosphorylatable serine, threonine and tyrosine residues. The kinetic data from peptide phosphorylation assays for approximately 330 oligopeptides is also included. The database entries are cross-referenced to the corresponding records in the Swiss-Prot protein database and literature references are linked to MedLine records. PhosphoBase is available via the WWW at http://www.cbs.dtu. dk/databases/PhosphoBase/ PMID- 9847190 TI - MMDB: Entrez's 3D structure database. AB - The three dimensional structures for representatives of nearly half of all protein families are now available in public databases. Thus, no matter which protein one investigates, it is increasingly likely that the 3D structure of a homolog will be known and may reveal unsuspected structure-function relationships. The goal of Entrez's 3D-structure database is to make this information accessible and usable by molecular biologists (http://www.ncbi.nlm.nih.gov/Entrez). To this end Entrez provides two major analysis tools, a search engine based on sequence and structure 'neighboring' and an integrated visualization system for sequence and structure alignments. From a protein's sequence 'neighbors' one may rapidly identify other members of a protein family, including those where 3D structure is known. By comparing aligned sequences and/or structures in detail, using the visualization system, one may identify conserved features and perhaps infer functional properties. Here we describe how these analysis tools may be used to investigate the structure and function of newly discovered proteins, using the PTEN gene product as an example. PMID- 9847191 TI - Protein folds and families: sequence and structure alignments. AB - Dali and HSSP are derived databases organizing protein space in the structurally known regions. We use an automatic structure alignment program (Dali) for the classification of all known 3D structures based on all-against-all comparison of 3D structures in the Protein Data Bank. The HSSP database associates 1D sequences with known 3D structures using a position-weighted dynamic programming method for sequence profile alignment (MaxHom). As a result, the HSSP database not only provides aligned sequence families, but also implies secondary and tertiary structures covering 36% of all sequences in Swiss-Prot. The structure classification by Dali and the sequence families in HSSP can be browsed jointly from a web interface providing a rich network of links between neighbours in fold space, between domains and proteins, and between structures and sequences. In particular, this results in a database of explicit multiple alignments of protein families in the twilight zone of sequence similarity. The organization of protein structures and families provides a map of the currently known regions of the protein universe that is useful for the analysis of folding principles, for the evolutionary unification of protein families and for maximizing the information return from experimental structure determination. The databases are available from http://www.embl-ebi.ac.uk/dali/ PMID- 9847192 TI - INFOGENE: a database of known gene structures and predicted genes and proteins in sequences of genome sequencing projects. AB - INFOGENE is a database of known and predicted gene structures with descriptions of basic functional signals and gene components. It provides a possibility to create compilations of sequences with a given gene feature as well as to accumulate and analyze predicted genes in finished and unfinished sequences from genome sequencing projects. Protein sequence similarity searches in the database of predicted proteins is offered through the BLASTP program. INFOGENE is realized under the Sequence Retrieval System that provides useful links with the other informational databases. The database is available through the WWW server of the Computational Genomics Group at http://genomic.sanger.ac.uk/db.html PMID- 9847193 TI - The PRESAGE database for structural genomics. AB - The PRESAGE database is a collaborative resource for structural genomics. It provides a database of proteins to which researchers add annotations indicating current experimental status, structural predictions and suggestions. The database is intended to enhance communication among structural genomics researchers and aid dissemination of their results. The PRESAGE database may be accessed at http://presage.stanford.edu/ PMID- 9847194 TI - SCOP: a Structural Classification of Proteins database. AB - The Structural Classification of Proteins (SCOP) database provides a detailed and comprehensive description of the relationships of all known proteins structures. The classification is on hierarchical levels: the first two levels, family and superfamily, describe near and far evolutionary relationships; the third, fold, describes geometrical relationships. The distinction between evolutionary relationships and those that arise from the physics and chemistry of proteins is a feature that is unique to this database, so far. The database can be used as a source of data to calibrate sequence search algorithms and for the generation of population statistics on protein structures. The database and its associated files are freely accessible from a number of WWW sites mirrored from URL http://scop. mrc-lmb.cam.ac.uk/scop/ PMID- 9847195 TI - The SBASE protein domain library, release 6.0: a collection of annotated protein sequence segments. AB - The sixth release of the SBASE protein domain library sequences contains 130 703 annotated and crossreferenced entries corresponding to structural, functional, ligand-binding and topogenic segments of proteins. The entries were grouped based on standard names (2312 groups) and futher classified on the basis of the BLAST similarity (2463 clusters). Automated searching with BLAST and a new sequence plot representation of local domain similarities are available at the WWW-server http://www.icgeb.trieste.it/sbase. A mirror site is at http://sbase.abc.hu/sbase. The database is freely available by anonymous 'ftp' file transfer from ftp.icgeb.trieste.it PMID- 9847196 TI - Pfam 3.1: 1313 multiple alignments and profile HMMs match the majority of proteins. AB - Pfam is a collection of multiple alignments and profile hidden Markov models of protein domain families. Release 3.1 is a major update of the Pfam database and contains 1313 families which are available on the World Wide Web in Europe at http://www.sanger.ac.uk/Software/Pfam/ and http://www.cgr.ki.se/Pfam/, and in the US at http://pfam.wustl.edu/. Over 54% of proteins in SWISS-PROT-35 and SP-TrEMBL 5 match a Pfam family. The primary changes of Pfam since release 2.1 are that we now use the more advanced version 2 of the HMMER software, which is more sensitive and provides expectation values for matches, and that it now includes proteins from both SP-TrEMBL and SWISS-PROT. PMID- 9847197 TI - Recent improvements of the ProDom database of protein domain families. AB - The ProDom database contains protein domain families generated from the SWISS PROT database by automated sequence comparisons. The current version was built with a new improved procedure based on recursive PSI-BLAST homology searches. ProDom can be searched on the World Wide Web to study domain arrangements within either known families or new proteins, with the help of a user-friendly graphical interface (http://www.toulouse.inra.fr/prodom.html). Recent improvements to the ProDom server include: ProDom queries under the SRS Sequence Retrieval System; links to the PredictProtein server; phylogenetic trees and condensed multiple alignments for a better representation of large domain families, with zooming in and out capabilities. In addition, a similar server was set up to display the outcome of whole genome domain analysis as applied to 17 completed microbial genomes (http://www.toulouse.inra.fr/prodomCG.html ). PMID- 9847198 TI - ISSD Version 2.0: taxonomic range extended. AB - Two more organisms from different taxonomic groups were added to a new version of the Integrated Sequence-Structure Database (ISSD). ISSD serves as an integrated source of sequence and structure information for the analysis of correlations between mRNA synonymous codon usage and three-dimensional structure of the encoded proteins. ISSD now holds 88 non-homologous Escherichia coli proteins and 25 yeast Saccharomyces cerevisiae proteins in addition to the expanded set of mammalian proteins, which includes 166 proteins (107 in ISSD Version 1.0). Comparison of ISSD sequences with organism-specific codon usage data derived from CUTG database shows that it is a representative subset of the GenBank coding sequences data. Preliminary results of the statistical analysis confirm that sequence-structure correlations observed by us earlier are also present in the upgraded ISSD (Version 2.0), including bacterial and yeast proteins. The ISSD Version 2.0 release includes an improved Web-based data search and retrieval system and is accessible via URL http://www.protein.bio.msu.su/issd/. ISSD can be also accessed at ExPASy, URL http://www.expasy.ch/swissmod/swiss-model.htm l PMID- 9847199 TI - ProClass Protein Family Database. AB - ProClass is a protein family database that organizes non-redundant sequence entries into families defined collectively by PROSITE patterns and PIR superfamilies. By combining global similarities and functional motifs into a single classification scheme, ProClass helps to reveal domain and family relationships and classify multi-domain proteins. The database currently consists of more than 120 000 sequence entries, approximately 60% of which is classified into about 3500 families. To maximize family information retrieval, the database provides links to various protein family/domain and structural class databases and contains multiple motif alignments of all PROSITE patterns as well as global alignments of PIR superfamilies. The motif sequences are retrieved from both PIR International and SWISS-PROT databases, including a large number of new members detected by our GeneFIND family identification system. ProClass can be used to support full-scale genomic annotation, because of its high classification rate. The ProClass database is available for on-line search and record retrieval from our WWW server at http://diana.uthct.edu/proclass.html PMID- 9847200 TI - The CATH Database provides insights into protein structure/function relationships. AB - We report the latest release (version 1.4) of the CATH protein domains database (http://www.biochem.ucl.ac.uk/bsm/cath). This is a hierarchical classification of 13 359 protein domain structures into evolutionary families and structural groupings. We currently identify 827 homologous families in which the proteins have both structual similarity and sequence and/or functional similarity. These can be further clustered into 593 fold groups and 32 distinct architectures. Using our structural classification and associated data on protein functions, stored in the database (EC identifiers, SWISS-PROT keywords and information from the Enzyme database and literature) we have been able to analyse the correlation between the 3D structure and function. More than 96% of folds in the PDB are associated with a single homologous family. However, within the superfolds, three or more different functions are observed. Considering enzyme functions, more than 95% of clearly homologous families exhibit either single or closely related functions, as demonstrated by the EC identifiers of their relatives. Our analysis supports the view that determining structures, for example as part of a 'structural genomics' initiative, will make a major contribution to interpreting genome data. PMID- 9847201 TI - Organising multi-dimensional biological image information: the BioImage Database. AB - Nowadays it is possible to unravel complex information at all levels of cellular organization by obtaining multi-dimensional image information. At the macromolecular level, three-dimensional (3D) electron microscopy, together with other techniques, is able to reach resolutions at the nanometer or subnanometer level. The information is delivered in the form of 3D volumes containing samples of a given function, for example, the electron density distribution within a given macromolecule. The same situation happens at the cellular level with the new forms of light microscopy, particularly confocal microscopy, all of which produce biological 3D volume information. Furthermore, it is possible to record sequences of images over time (videos), as well as sequences of volumes, bringing key information on the dynamics of living biological systems. It is in this context that work on BioImage started two years ago, and that its first version is now presented here. In essence, BioImage is a database specifically designed to contain multi-dimensional images, perform queries and interactively work with the resulting multi-dimensional information on the World Wide Web, as well as accomplish the required cross-database links. Two sister home pages of BioImage can be accessed at http://www. bioimage.org and http://www-embl.bioimage.org PMID- 9847202 TI - Database of protein sequence alignments: PIR-ALN. AB - The Protein Information Resource (PIR) has been maintaining a database of curated protein sequence alignments since 1991. The collection includes superfamily, family and homology domain alignments. CLUSTAL V/W is used to generate multiple sequence alignments and ALNED, an interactive alignment editor, is used to check and correct them. The database has helped in classifying sequences, in defining new homology domains, and in spreading and standardizing protein names, features and keywords among members of a family or superfamily. The ATLAS information retrieval system can be used to browse and query the PIR-ALN alignments. The quarterly and weekly updates can be accessed via the WWW at http://www-nbrf. georgetown.edu/pir/ PMID- 9847203 TI - ProTherm: Thermodynamic Database for Proteins and Mutants. AB - The first release of the Thermodynamic Database for Proteins and Mutants (ProTherm) contains more than 3300 data of several thermodynamic parameters for wild type and mutant proteins. Each entry includes numerical data for unfolding Gibbs free energy change, enthalpy change, heat capacity change, transition temperature, activity etc., which are important for understanding the mechanism of protein stability. ProTherm also includes structural information such as secondary structure and solvent accessibility of wild type residues, and experimental methods and other conditions. A WWW interface enables users to search data based on various conditions with different sorting options for outputs. Further, ProTherm is cross-linked with NCBI PUBMED literature database, Protein Mutant Database, Enzyme Code and Protein Data Bank structural database. Moreover, all the mutation sites associated with each PDB structure are automatically mapped and can be directly viewed through 3DinSight developed in our laboratory. The database is available at the URL, http://www.rtc.riken.go.jp/protherm.htm l PMID- 9847204 TI - The SWISS-2DPAGE database: what has changed during the last year. AB - SWISS-2DPAGE (http://www.expasy.ch/ch2d/) is an annotated two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) database established in 1993. The current release contains 21 reference maps from human and mouse biological samples, as well as from Saccharomyces cerevisiae, Escherichia coli and Dictyostelium discoideum origin. These reference maps now have 2480 identified spots, corresponding to 528 separate protein entries in the database, in addition to virtual entries for each SWISS-PROT sequence. During the last year, the SWISS 2DPAGE has undergone major changes. Six new maps have been added, and new functions to access the data have been provided through the ExPASy server. Finally, an important change concerns the database funding source. PMID- 9847205 TI - Codon usage tabulated from the international DNA sequence databases; its status 1999. AB - Frequencies for each of the 206 526 complete protein-coding genes (CDS's) have been compiled from taxonomical divisions of the GenBank DNA sequence database. The sum of the codon use of 7434 organisms has also been calculated. These data files can be obtained from anonymous ftp sites of DDBJ, DISC and EBI. The list of the codon usage of genes in an organism as well as the sum of the codon usage of the organism was made searchable by the name of organism through a web site http://www.dna.affrc.go.jp//CUTG.html PMID- 9847206 TI - TransTerm, the translational signal database, extended to include full coding sequences and untranslated regions. AB - TransTerm is a database of mRNA sequences and parameters useful for detecting translational control signals in general. TransTerm-98 has been expanded beyond previous years to include full coding sequences and UTRs, while retaining the original small contexts about the coding sequence start- and stop-codons. The database contains more than 130 000 non-redundant coding sequences with associated untranslated regions (UTRs) from over 450 species. This includes the complete genomes of 12 prokaryotic and one eukaryotic organism. Several coding sequence parameters are available: coding sequence length, Nc, GC3 and, when it is computable, Codon Adaptation Index (CAI). Codon usage tables and summaries of start- and stop-codon contexts are also included. TransTerm-98 has both a relational database form with a WWW interface and a flatfile format, also available by Internet browser. TransTerm is available at: http://biochem.otago.ac.nz:800/Transterm/homepage.h tml PMID- 9847207 TI - PlantCARE, a plant cis-acting regulatory element database. AB - PlantCARE is a database of plant cis- acting regulatory elements, enhancers and repressors. Besides the transcription motifs found on a sequence, it also offers a link to the EMBL entry that contains the full gene sequence as well as a description of the conditions in which a motif becomes functional. The information on these sites is given by matrices, consensus and individual site sequences on particular genes, depending on the available information. PlantCARE is a relational database available via the web at the URL: http://sphinx.rug.ac.be:8080/PlantCARE/ PMID- 9847208 TI - Plant cis-acting regulatory DNA elements (PLACE) database: 1999. AB - PLACE (http://www.dna.affrc.go.jp/htdocs/PLACE/) is a database of nucleotide sequence motifs found in plant cis-acting regulatory DNA elements. Motifs were extracted from previously published reports on genes in vascular plants. In addition to the motifs originally reported, their variations in other genes or in other plant species in later reports are also compiled. Documents for each motif in the PLACE database contains, in addition to a motif sequence, a brief definition and description of each motif, and relevant literature with PubMed ID numbers and GenBank accession numbers where available. Users can search their query sequences for cis-elements using the Signal Scan program at our web site. The results will be reported in one of the three forms. Clicking the PLACE accession numbers in the result report will open the pertinent motif document. Clicking the PubMed or GenBank accession number in the document will allow users to access to these databases, and to read the of the literature or the annotation in the DNA database. This report summarizes the present status of this database and available tools. PMID- 9847209 TI - ASDB: database of alternatively spliced genes. AB - A database of alternatively spliced genes (ASDB) has been constructed based on (i) the results of the analysis of Swiss-Prot entries containing products of these genes and (ii) clustering procedure joining proteins that could arise by alternative splicing of the same gene. ASDB incorporates information about alternatively spliced genes, their products and expression patterns. It can be searched in order to find all products of alternative splicing produced in a particular tissue or a given organism, or all variants generated by a particular transcript. ASDB currently contains about 1700 protein sequences and can be accessed via the Internet at URL http://cbcg.nersc.gov/asdb PMID- 9847210 TI - Transcription Regulatory Regions Database (TRRD):its status in 1999. AB - The Transcription Regulatory Regions Database (TRRD) is a curated database designed for accumulation of experimental data on extended regulatory regions of eukaryotic genes, the regulatory elements they contain, i.e., transcription factor binding sites, promoters, enhancers, silencers, etc., and expression patterns of the genes. Release 4.1 of TRRD offers a number of significant improvements, in particular, a more detailed description of transcription factor binding sites, transcription factors per se, and gene expression patterns in a computer-readable format. In addition, the new TRRD release provides considerably more references to other molecular biological databases. TRRD 4.1 is installed under SRS and is available through the WWW at http://www.bionet.nsc.ru/trrd/ PMID- 9847212 TI - The ENZYME data bank in 1999. AB - The ENZYME data bank is a repository of information related to the nomenclature of enzymes. In recent years it has become an indispensable resource for the development of metabolic databases. The current version contains information on 3704 enzymes. It is available through the ExPASy WWW server (http://www.expasy.ch/). PMID- 9847211 TI - The Eukaryotic Promoter Database (EPD): recent developments. AB - The Eukaryotic Promoter Database (EPD) is an annotated non-redundant collection of eukaryotic POL II promoters, for which the transcription start site has been determined experimentally. Access to promoter sequences is provided by pointers to positions in nucleotide sequence entries. The annotation part of an entry includes description of the initiation site mapping data, cross-references to other databases, and bibliographic references. EPD is structured in a way that facilitates dynamic extraction of biologically meaningful promoter subsets for comparative sequence analysis. Recent efforts have focused on exhaustive cross referencing to the EMBL nucleotide sequence database, and on the improvement of the WWW-based user interfaces and data retrieval mechanisms. EPD can be accessed at http://www.epd.isb-sib.ch PMID- 9847213 TI - REBASE-restriction enzymes and methylases. AB - REBASE is a comprehensive database of information about restriction enzymes and their associated methylases, including their recognition and cleavage sites and their commercial availability. Also included is a listing of homing endonucleases. Information from REBASE is distributed via monthly electronic mailings as well as through anonymous ftp and the World Wide Web. The REBASE web site (http://www.neb.com/rebase) contains a web page for every enzyme, reference and supplier. Additionally, there is a search facility, help and NEWS pages, and a complete description of our various services. Specialized files are available that can be used directly by many software packages. PMID- 9847214 TI - The Ribonuclease P Database. AB - Ribonuclease P is responsible for the 5'-maturation of tRNA precursors. Ribonuclease P is a ribonucleoprotein, and in bacteria (and some Archaea) the RNA subunit alone is catalytically active in vitro, i.e. it is a ribozyme. The Ribonuclease P Database is a compilation of ribonuclease P sequences, sequence alignments, secondary structures, three-dimensional models and accessory information, available via the World Wide Web at the following URL: http://www.mbio.ncsu.edu/RNaseP/home .html PMID- 9847215 TI - Object oriented Transcription Factors Database (ooTFD). AB - ooTFD is an object-oriented database for the representation of information pertaining to transcription factors, the proteins and biochemical entities which play a central role in the regulation of gene expression. Given the recent explosion of genome sequence information, and that a large percentage of proteins encoded by fully sequenced genomes fall into this category, information pertaining to this class of molecules may become an essential aspect of biology and of genomics in the 21st century. In the past year, there was a small increase in the size of this database, and a number of new tools to facilitate data access and analysis have been added at the MIRAGE (Molecular Informatics Resource for the Analysis of Gene Expression) web site. ooTFD and associated tools and resources can be accessed at http://www.ifti.org/ PMID- 9847217 TI - Histone Sequence Database: sequences, structures, post-translational modifications and genetic loci. AB - The Histone Sequence Database is an annotated and searchable collection of all available histone and histone fold sequences and structures. Particular emphasis has been placed on documenting conflicts between similar sequence entries from a number of source databases, conflicts that are not necessarily documented in the source databases themselves. New additions to the database include compilations of post-translational modifications for each of the core and linker histones, as well as genomic information in the form of map loci for the human histone gene complement, with the genetic loci linked to Online Mendelian Inheritance in Man (OMIM). The database is freely accessible through the World Wide Web at either http://genome.nhgri.nih.gov/histones/ or http://www.ncbi.nlm.nih. gov/Baxevani/HISTONES PMID- 9847216 TI - Expanding the TRANSFAC database towards an expert system of regulatory molecular mechanisms. AB - TRANSFAC is a database on transcription factors, their genomic binding sites and DNA-binding profiles. In addition to being updated and extended by new features, it has been complemented now by a series of additional database modules. Among them, modules which provide data about signal transduction pathways (TRANSPATH) or about cell types/organs/developmental stages (CYTOMER) are available as well as an updated version of the previously described COMPEL database. The databases are available on the WWW at http://transfac.gbf.de/ PMID- 9847218 TI - MEROPS: the peptidase database. AB - The MEROPS database (http://www.bi.bbsrc.ac.uk/Merops/Merops.+ ++htm) provides a catalogue and structure-based classification of peptidases (i.e. all proteolytic enzymes). This is a large group of proteins (approximately 2% of all gene products) that is of particular importance in medicine and biotechnology. An index of the peptidases by name or synonym gives access to a set of files termed PepCards each of which provides information on a single peptidase. Each card file contains information on classification and nomenclature, and hypertext links to the relevant entries in online databases for human genetics, protein and nucleic acid sequence data and tertiary structure. Another index provides access to the PepCards by organism name so that the user can retrieve all known peptidases from a particular species. The peptidases are classified into families on the basis of statistically significant similarities between the protein sequences in the part termed the 'peptidase unit' that is most directly responsible for activity. Families that are thought to have common evolutionary origins and are known or expected to have similar tertiary folds are grouped into clans. The MEROPS database provides sets of files called FamCards and ClanCards describing the individual families and clans. Each FamCard document provides links to other databases for sequence motifs and secondary and tertiary structures, and shows the distribution of the family across the major kingdoms of living creatures. Release 3.03 of MEROPS contains 758 peptidases, 153 families and 22 clans. We suggest that the MEROPS database provides a model for a way in which a system of classification for a functional group of proteins can be developed and used as an organizational framework around which to assemble a variety of related information. PMID- 9847219 TI - The aminoacyl-tRNA Synthetase Data Bank (AARSDB). AB - Aminoacyl-tRNA synthetases (AARSs) are the key components of the protein biosynthesis machinery. They are responsible for maintaining the fidelity of transfer of genetic information from DNA into protein. The database is a compilation of amino acid sequences of all aminoacyl-tRNA synthetases known to date. It contains 422 primary structures of the AARSs available as separate entries or alignments of related proteins. The database is available via the World Wide Web at http://rose.man.poznan.pl/aars/index.html PMID- 9847220 TI - The Homeodomain Resource: sequences, structures and genomic information. AB - The Homeodomain Resource is a comprehensive collection of sequence, structure and genomic information on the homeodomain protein family. Available through the Resource are both full-length and domain-only sequence data, as well as X-ray and NMR structural data for proteins and protein-DNA complexes. Also available is information on human genetic diseases and disorders in which proteins from the homeodomain family play an important role; genomic information includes relevant gene symbols, cytogenetic map locations, and specific mutation data. Search engines are provided to allow users to easily query the component databases and assemble specialized data sets. The Homeodomain Resource is available through the World Wide Web at http://genome.nhgri.nih.gov/homeodomain PMID- 9847221 TI - HUGE: a database for human large proteins identified by Kazusa cDNA sequencing project. AB - HUGE is a database for human large proteins newly identified by Kazusa cDNA project, which aims to predict protein primary structures from sequences of human large cDNAs (>4 kb). In particular, cDNA clones capable of coding for large proteins (>50 kDa) are current targets of the project. More than 700 sequences of human cDNAs (average size, 5.1 kb) have been determined to date and deposited in the public databases. Notable information implied from the cDNAs and the predicted protein sequences can be obtained through HUGE via the World Wide Web at URL http://www.kazusa.or.jp/huge PMID- 9847223 TI - Olfactory Receptor Database: a database of the largest eukaryotic gene family. AB - The Olfactory Receptor Database (ORDB) is a WWW-accessible database that stores data on Olfactory Receptor-like molecules (ORs) and has been open to the public since June 1996. It contains a public and a private area. The public area includes published DNA and protein sequence data for ORs, links to OR models and data on their expression, chromosomal localization and source organism, as well as (i) links to bibliography through PubMed and (ii) interactive WWW-based tools, such as BLAST homology searching. The private area functions as a service to laboratories that are actively cloning receptors. Source laboratories enter the sequences of the receptor clones they have characterized to the private database and can search for identical or near identical OR sequences in both public and private databases. If another laboratory has cloned and deposited an identical or closely matching sequence there are means for communication between the laboratories to help avoid duplication of work. ORDB is available via the WWW at http://crepe.med.yale.edu/ORDB/HTML PMID- 9847222 TI - The Ligand Gated Ion Channel Database. AB - The ligand gated ion channels (LGICs) are ionotropic receptors to neurotransmitters. Their physiological effect is carried out by the opening of an ionic channel upon binding of a particular neurotransmitter. These LGICs constitute superfamilies of receptors formed by homologous subunits. A database has been developed to handle the growing wealth of cloned subunits. This database contains nucleic acid sequences, protein sequences, as well as multiple sequence alignments and phylogenetic studies. This database is accessible via the worldwide web (http://www.pasteur.fr/units/neubiomol/LGIC.h tml), where it is continuously updated. A downloadable version is also available [currently v0.1 (98.06)]. PMID- 9847224 TI - InBase, the New England Biolabs Intein Database. AB - Inteins are intervening sequences that splice as proteins, not RNA. InBase, the New England Biolabs Intein Database (http://www.neb. com/neb/inteins.html), is a comprehensive on-line database that includes the Intein Registry, along with detailed information about each intein and its host protein, tabulated comparisons and a comprehensive bibliography including papers in press. PMID- 9847225 TI - Human Immunodeficiency Virus Reverse Transcriptase and Protease Sequence Database. AB - The HIV RT and Protease Sequence Database is an on-line relational database that catalogues evolutionary and drug-related human immunodeficiency virus reverse transcriptase (RT) and protease sequence variation (http://hivdb.stanford.edu). The database contains a compilation of nearly all published HIV RT and protease sequences including International Collaboration database submissions (e.g., GenBank) and sequences published in journal articles. Sequences are linked to data about the source of the sequence sample and the anti-HIV drug treatment history of the individual from whom the isolate was obtained. The database is curated and sequences are annotated with data from 180 literature references. Users can retrieve additional data and view alignments of sequences sets meeting specific criteria (e.g., treatment history, subtype, presence of a particular mutation). PMID- 9847226 TI - A database on cytogenetics in haematology and oncology. AB - The aim of 'Atlas of Genetics and Cytogenetics in Oncology and Haematology' (http://www.infobiogen.fr/services/chromcancer) is to present summarized information on chromosome abnormalities in cancer, with extensions to genes involved in cancer and to cancer-prone diseases. Information is to be updated. This database is made for and by cytogeneticists, molecular biologists, clinicians in oncology and in haematology, and pathologists. PMID- 9847227 TI - The Protein Mutant Database. AB - Currently the protein mutant database (PMD) contains over 81 000 mutants, including artificial as well as natural mutants of various proteins extracted from about 10 000 articles. We recently developed a powerful viewing and retrieving system (http://pmd.ddbj.nig.ac.jp), which is integrated with the sequence and tertiary structure databases. The system has the following features: (i) mutated sequences are displayed after being automatically generated from the information described in the entry together with the sequence data of wild-type proteins integrated. This is a convenient feature because it allows one to see the position of altered amino acids (shown in a different color) in the entire sequence of a wild-type protein; (ii) for those proteins whose 3D structures have been experimentally determined, a 3D structure is displayed to show mutation sites in a different color; (iii) a sequence homology search against PMD can be carried out with any query sequence; (iv) a summary of mutations of homologous sequences can be displayed, which shows all the mutations at a certain site of a protein, recorded throughout the PMD. PMID- 9847228 TI - Keio Mutation Database for eye disease genes (KMeyeDB). AB - A database of mutations in human eye disease genes has been constructed. This KMeyeDB employs a database software MutationView which provides graphical data presentation and analysis as a smooth user-interface. Currently, the KMeyeDB contains mutation data of 16 different genes for 18 eye diseases. The KMeyeDB is accessible through http://mutview.dmb.med.keio.ac.jp with advanced internet browsers. PMID- 9847229 TI - KinMutBase, a database of human disease-causing protein kinase mutations. AB - KinMutBase (http://www.uta.fi/laitokset/imt/KinMut Base.html) is a registry of mutations in human protein kinases related to disorders. Kinases are essential cellular signalling molecules, in which mutations can lead into diseases including, e.g., immunodeficiencies, cancers and endocrine disorders. The first release of KinMutBase contains information for nine protein tyrosine kinases. There are altogether 170 entries representing 273 families and 403 patients. Mutations appear both in conserved hallmark residues of the kinases as well as in non-homologous sites. The KinMutBase WWW pages provide plenty of information, namely mutation statistics and display, clickable sequences with mutations, restriction enzyme patterns and online submission. PMID- 9847230 TI - KEYnet: a keywords database for biosequences functional organization. AB - KEYnet is a database where gene and protein names are hierarchically structured. Particular care has been devoted to the search and organisation of synonyms. The structuring is based on biological criteria in order to assist the user in the data search and to minimise the risk of loss of information. Links to the EMBL data library by the entry name and the accession number have been implemented. KEYnet is available through the World Wide Web at the following site: http://www.ba.cnr.it/keynet.html. Recently KEYnet has incorporated specific gene name classifications, which can be browsed starting from the above-mentioned KEYnet home page: the Mitochondrial Gene Names classification and the Rat Gene Names classification. KEYnet database has also been structured in a flatfile format and can be queried through SRS (http://bio-www.ba.cnr.t:8000/srs). PMID- 9847231 TI - AAindex: Amino Acid Index Database. AB - AAindex is a database of numerical indices representing various physicochemical and biochemical properties of amino acids and pairs of amino acids. It consists of two sections: AAindex1 for the amino acid index of 20 numerical values and AAindex2 for the amino acid mutation matrix of 210 numerical values. Each entry of either AAindex1 or AAindex2 consists of the definition, the reference information, a list of related entries in terms of the correlation coefficient, and the actual data. The database may be accessed through the DBGET/LinkDB system at GenomeNet (http://www.genome.ad. jp/dbget/) or may be downloaded by anonymous FTP (ftp://ftp.genome. ad.jp/db/genomenet/aaindex/). PMID- 9847232 TI - O-GLYCBASE version 4.0: a revised database of O-glycosylated proteins. AB - O-GLYCBASE is a database of glycoproteins with O-linked glycosylation sites. Entries with at least one experimentally verified O-glycosylation site have been compiled from protein sequence databases and literature. Each entry contains information about the glycan involved, the species, sequence, a literature reference and http-linked cross-references to other databases. Version 4.0 contains 179 protein entries, an approximate 15% increase over the last version. Sequence logos representing the acceptor specificity patterns for GalNAc, GlcNAc, mannosyl and xylosyl transferases are shown. The O-GLYCBASE database is available through the WWW at http://www.cbs.dtu.dk/databases/OGLYCBASE/ PMID- 9847234 TI - LIGAND database for enzymes, compounds and reactions. AB - LIGAND is a composite database consisting of three sections and containing the information of chemical substances, chemical reactions and enzymes that catalyze reactions. The COMPOUND section is a collection of metabolic compounds, as well as macromolecules, chemical elements and other chemical substances in a living cell. The ENZYME section is a collection of all known enzymatic reactions, together with the information of enzyme molecules, classified according to the EC (Enzyme Commission) numbers. The REACTION section is a new addition to the database containing metabolic reactions that appear in the pathway diagrams of the KEGG/PATHWAY database and/or in the ENZYME section. The LIGAND database can be accessed through the WWW (http://www.genome.ad.jp/dbget/ligand.html) or may be downloaded by anonymous FTP (ftp://kegg.genome.ad. jp/molecules/ligand). PMID- 9847233 TI - The University of Minnesota Biocatalysis/Biodegradation Database: specialized metabolism for functional genomics. AB - The University of Minnesota Biocatalysis/Biodegradation Database (UM-BBD, http://www.labmed.umn.edu/umbbd/i nde x.html) first became available on the web in 1995 to provide information on microbial biocatalytic reactions of, and biodegradation pathways for, organic chemical compounds, especially those produced by man. Its goal is to become a representative database of biodegradation, spanning the diversity of known microbial metabolic routes, organic functional groups, and environmental conditions under which biodegradation occurs. The database can be used to enhance understanding of basic biochemistry, biocatalysis leading to speciality chemical manufacture, and biodegradation of environmental pollutants. It is also a resource for functional genomics, since it contains information on enzymes and genes involved in specialized metabolism not found in intermediary metabolism databases, and thus can assist in assigning functions to genes homologous to such less common genes. With information on >400 reactions and compounds, it is poised to become a resource for prediction of microbial biodegradation pathways for compounds it does not contain, a process complementary to predicting the functions of new classes of microbial genes. PMID- 9847235 TI - A new family of growth factors produced by the fat body and active on Drosophila imaginal disc cells. AB - By fractionating conditioned medium (CM) from Drosophila imaginal disc cell cultures, we have identified a family of Imaginal Disc Growth Factors (IDGFs), which are the first polypeptide growth factors to be reported from invertebrates. The active fraction from CM, as well as recombinant IDGFs, cooperate with insulin to stimulate the proliferation, polarization and motility of imaginal disc cells. The IDGF family in Drosophila includes at least five members, three of which are encoded by three genes in a tight cluster. The proteins are structurally related to chitinases, but they show an amino acid substitution that is known to abrogate catalytic activity. It therefore seems likely that they have evolved from chitinases but acquired a new growth-promoting function. The IDGF genes are expressed most strongly in the embryonic yolk cells and in the fat body of the embryo and larva. The predicted molecular structure, expression patterns, and mitogenic activity of these proteins suggest that they are secreted and transported to target tissues via the hemolymph. However, the genes are also expressed in embryonic epithelia in association with invagination movements, so the proteins may have local as well as systemic functions. Similar proteins are found in mammals and may constitute a novel class of growth factors. PMID- 9847236 TI - Conserved regulation of mesenchymal gene expression by Fgf-8 in face and limb development. AB - Clim-2 (NLI, Lbd1) is one of two related mouse proteins that interact with Lim domain homeoproteins. In the mouse, embryonic expression of Clim-2 is particularly pronounced in facial ectomesenchyme and limb bud mesenchyme in association with Lim genes, Lhx-6 and Lmx-1 respectively. We show that in common with both these Lim genes, Clim-2 expression is regulated by signals from overlying epithelium. In both the developing face and the limb buds we identify Fgf-8 as the likely candidate signalling molecule that regulates Clim-2 expression. We show that in the mandibular arch, as in the limb, Fgf-8 functions in combination with CD44, a cell surface binding protein, and that blocking CD44 binding results in inhibition of Fgf8-induced expression of Clim-2 and Lhx-6. Regulation of gene expression by Fgf8 in association with CD44 is thus conserved between limb and mandibular arch development. PMID- 9847237 TI - Antivin, a novel and divergent member of the TGFbeta superfamily, negatively regulates mesoderm induction. AB - Mesoderm induction and patterning are mediated by members of the TGFbeta superfamily. We have isolated a novel zebrafish member, antivin, that structurally is highly related to mouse lefty. Overexpression of antivin completely abolishes mesoderm induction at blastula stage, yet resultant embryos develop well-patterned epidermal and neural derivatives. The mesoderm-inhibiting activity of antivin can be mimicked by lefty and is suppressed by increasing levels of the mesodermal inducer Activin or its receptors. On the basis of its expression and activity, we propose that Antivin normally functions as a competitive inhibitor of Activin to limit mesoderm induction in the early embryo. PMID- 9847238 TI - A BMP homolog acts as a dose-dependent regulator of body size and male tail patterning in Caenorhabditis elegans. AB - We cloned the dbl-1 gene, a C. elegans homolog of Drosophila decapentaplegic and vertebrate BMP genes. Loss-of-function mutations in dbl-1 cause markedly reduced body size and defective male copulatory structures. Conversely, dbl-1 overexpression causes markedly increased body size and partly complementary male tail phenotypes, indicating that DBL-1 acts as a dose-dependent regulator of these processes. Evidence from genetic interactions indicates that these effects are mediated by a Smad signaling pathway, for which DBL-1 is a previously unidentified ligand. Our study of the dbl-1 expression pattern suggests a role for neuronal cells in global size regulation as well as male tail patterning. PMID- 9847239 TI - Specificity of TGFbeta signaling is conferred by distinct type I receptors and their associated SMAD proteins in Caenorhabditis elegans. AB - In C. elegans, the TGFbeta-like type II receptor daf-4 is required for two distinct signaling pathways. In association with the type I receptor daf-1, it functions in the dauer pathway. In addition, it is also required for body size determination and male tail patterning, roles which do not require daf-1. In an effort to determine how two different signals are transmitted through daf-4, we looked for other potential signaling partners for DAF-4. We have cloned and characterized a novel type I receptor and show that it is encoded by sma-6. Mutations in sma-6 generate the reduced body size (Sma) and abnormal mail tail (Mab) phenotypes identical to those observed in daf-4 and sma-2, sma-3, sma-4 mutants (C. elegans Smads), indicating that they function in a common signaling pathway. However, mutations in sma-6, sma-2, sma-3, or sma-4 do not produce constitutive dauers, which demonstrates that the unique biological functions of daf-4 are mediated by distinct type I receptors functioning in parallel pathways. We propose that the C. elegans model for TGFbeta-like signaling, in which distinct type I receptors determine specificity, may be a general mechanism of achieving specificity in other organisms. These findings distinguish between the manner in which signaling specificity is achieved in TGFbeta-like pathways and receptor tyrosine-kinase (RTK) pathways. PMID- 9847240 TI - Fasciclin II and Beaten path modulate intercellular adhesion in Drosophila larval visual organ development. AB - Previous studies demonstrated that Fasciclin II and Beaten path are necessary for regulating cell adhesion events that are important for motoneuron development in Drosophila. We observe that the cell adhesion molecule Fasciclin II and the secreted anti-adhesion molecule Beaten path have additional critical roles in the development of at least one set of sensory organs, the larval visual organs. Taken together, phenotypic analysis, genetic interactions, expression studies and rescue experiments suggest that, in normal development, secretion of Beaten path by cells of the optic lobes allows the Fasciclin II-expressing larval visual organ cells to detach from the optic lobes as a cohesive cell cluster. Our results also demonstrate that mechanisms guiding neuronal development may be shared between motoneurons and sensory organs, and provide evidence that titration of adhesion and anti-adhesion is critical for early steps in development of the larval visual system. PMID- 9847241 TI - Laser ablation of persistent twist cells in Drosophila: muscle precursor fate is not segmentally restricted. AB - In Drosophila the precursors of the adult musculature arise during embryogenesis. These precursor cells have been termed Persistent Twist Cells (PTCs), as they continue to express the transcription factor Twist after that gene ceases expression elsewhere in the mesoderm. In the larval abdomen, the PTCs are associated with peripheral nerves in stereotypic ventral, dorsal, and lateral clusters, which give rise, respectively, to the ventral, dorsal, and lateral muscle fiber groups of the adult. We tested the developmental potential of the PTCs by using a microbeam laser to ablate specific clusters in larvae. We found that the ablation of a single segmental PTC cluster does not usually result in the deletion of the corresponding adult fibers of that segment. Instead, normal or near normal numbers of adult fibers can form after the ablation. Examination of pupae following ablation showed that migrating PTCs from adjacent segments are able to invade the affected segment, replenishing the ablated cells. However, the ablation of homologous PTCs in multiple segments does result in the deletion of the corresponding adult muscle fibers. These data indicate that the PTCs in an abdominal segment can contribute to the formation of muscle fibers in adjacent abdominal segments, and thus are not inherently restricted to the formation of muscle fibers within their segment of origin. PMID- 9847242 TI - Regionalization of Sonic hedgehog transcription along the anteroposterior axis of the mouse central nervous system is regulated by Hnf3-dependent and -independent mechanisms. AB - The axial midline mesoderm and the ventral midline of the neural tube, the floor plate, share the property of being a source of the secreted protein, Sonic hedgehog (Shh), which has the capacity to induce a variety of ventral cell types along the length of the mouse CNS. To gain insight into the mechanisms by which Shh transcription is initiated in these tissues, we set out to identify the cis acting sequences regulating Shh gene expression. As an approach, we have tested genomic clones encompassing 35 kb of the Shh locus for their ability to direct a lacZ reporter gene to the temporally and spatially restricted confines of the Shh expression domains in transgenic mice. Three enhancers were identified that directed lacZ expression to distinct regions along the anteroposterior axis including the ventral midline of the spinal cord, hindbrain, rostral midbrain and caudal diencephalon, suggesting that multiple transcriptional regulators are required to initiate Shh gene expression within the CNS. In addition, regulatory sequences were also identified that directed reporter expression to the notochord, albeit, under limited circumstances. Sequence analysis of the genomic clones responsible for enhancer activity from a variety of organisms, including mouse, chicken and human, have identified highly conserved binding sites for the hepatocyte nuclear factor 3 (Hnf3) family of transcriptional regulators in some, but not all, of the enhancers. Moreover, the generation of mutations in the Hnf3 binding sites showed their requirement in certain, but not all, aspects of Shh reporter expression. Taken together, our results support the existence of Hnf3 dependent and -independent mechanisms in the direct activation of Shh transcription within the CNS and axial mesoderm. PMID- 9847243 TI - The endocycle controls nurse cell polytene chromosome structure during Drosophila oogenesis. AB - Polytene chromosomes exhibit intricate higher order chromatin structure that is easily visualized due to their precisely aligned component strands. However, it remains unclear if the same factors determine chromatin organization in polyploid and diploid cells. We have analyzed one such factor, the cell cycle, by studying changes in Drosophila nurse cell chromosomes throughout the 10 to 12 endocycles of oogenesis. We find that nurse cells undergo three distinct types of endocycle whose parameters are correlated with chromosome behavior. The first four endocycles support complete DNA replication; poorly banded polytene euchromatin progressively condenses during the late S phases to produce blob-like chromosomes. During the unique fifth endocycle, an incomplete late S phase is followed by a mitosis-like state during which the 64C chromosomes dissociate into 32 chromatid pairs held together by unreplicated regions. All the subsequent endocycles lack any late S phase; during these cycles a new polytene chromosome grows from each 2C chromatid pair to generate 32-ploid polytene nuclei. These observations suggest that euchromatin begins to condense during late S phase and that nurse cell polytene chromosome structure is controlled by regulating whether events characteristic of late S and M phase are incorporated or skipped within a given endocycle. PMID- 9847244 TI - Mosaic analysis of the dominant mutant, Gnarley1-R, reveals distinct lateral and transverse signaling pathways during maize leaf development. AB - Maize leaves are organized into two major domains along the proximal-distal axis: a broad flat blade at the distal end of the leaf, and a narrow, thickened sheath that encircles the stem. Between the blade and sheath are two wedge-shaped tissues called auricles, and the ligule, an epidermally derived fringe. Members of the Knotted1 (Kn1) family of mutations change the shape and position of both ligule and auricle, thus disturbing the overall pattern of the leaf. Here we present the results of a mosaic analysis of Gnarley1-R (Gn1-R), which like members of the Kn1 family, affects the ligule and auricle. Gn1-R is distinct, however, in altering the dimensions of cells that make up sheath tissue. To gain insight into the Gn1-R phenotype, we performed a mosaic analysis using X-ray induced chromosome breakage to generate wild-type (gn1+/-) sectors in otherwise Gn1-R leaves. These sectors allowed us to determine whether Gn1-R acts non autonomously to influence adjacent cells. Most aspects of the Gn1-R phenotype, such as ligule position, inhibition of auricle development, and sheath thickness showed autonomy in the lateral dimension (leaf width). In contrast, all aspects of the Gn1-R phenotype were non-autonomous in the transverse dimension (leaf thickness), suggesting that signaling occurs between cell layers in the leaf. These results support a model for distinct signaling pathways along lateral versus transverse axes of a developing leaf. PMID- 9847245 TI - Control of phyllotaxy in maize by the abphyl1 gene. AB - Organogenesis in plants occurs at the shoot apical meristem, a group of indeterminate stem cells that are organized during embryogenesis. Regulated initiation of leaves or flowers from the shoot meristem gives rise to the familiar geometric patterns observed throughout the plant kingdom. The mechanism by which these patterns, termed phyllotaxies, are generated, remains unclear. Maize plants initiate leaves singly, alternating from one side to the other in a regular pattern. Here we describe a recessive maize mutant, abphyl1, that initiates leaves in opposite pairs, in a pattern termed decussate phyllotaxy. The decussate shoot meristems are larger than normal throughout development, though the general structure and organization of the meristem is not altered. abph1 mutants are first distinguished during embryogenesis, prior to true leaf initiation, by a larger shoot meristem and coincident larger expression domain of the homeobox gene knotted1. Therefore, the abph1 gene regulates morphogenesis in the embryo, and plays a role in determining the phyllotaxy of the shoot. PMID- 9847246 TI - Glycogen synthase kinase-3 (GSK-3) is regulated during Dictyostelium development via the serpentine receptor cAR3. AB - Glycogen synthase kinase-3 (GSK-3) is required during metazoan development to mediate the effects of the extracellular signal wingless/Wnt-1 and hence is necessary for correct cell type specification. GSK-3 also regulates cell fate during Dictyostelium development, but in this case it appears to mediate the effects of extracellular cAMP. By direct measurement of GSK-3 kinase activity during Dictyostelium development, we find that there is a rise in activity at the initiation of multicellular development which can be induced by cAMP. The timing of the rise correlates with the requirement for the Dictyostelium homologue of GSK-3, GSKA, to specify cell fate. We show that loss of the cAMP receptor cAR3 almost completely abolishes the rise in kinase activity and causes a mis specification of cell fate that is equivalent to that seen in a gskA- mutant. The phenotype of a cAR3(-) mutant however is less severe than loss of gskA and ultimately gives rise to an apparently wild-type fruiting body. These results indicate that in Dictyostelium extracellular cAMP acts via cAR3 to cause a rise in GSKA kinase activity which regulates cell type patterning during the initial stages of multicellularity. PMID- 9847247 TI - Signaling through the stromal epidermal growth factor receptor is necessary for mammary ductal development. AB - Stromal-epithelial interactions are critical in determining patterns of growth, development and ductal morphogenesis in the mammary gland, and their perturbations are significant components of tumorigenesis. Growth factors such as epidermal growth factor (EGF) contribute to these reciprocal stromal-epithelial interactions. To determine the role of signaling through the EGF receptor (EGFR) in mammary ductal growth and branching, we used mice with a targeted null mutation in the Egfr. Because Egfr-/- mice die perinatally, transplantation methods were used to study these processes. When we transplanted neonatal mammary glands under the renal capsule of immuno-compromised female mice, we found that EGFR is essential for mammary ductal growth and branching morphogenesis, but not for mammary lobulo-alveolar development. Ductal growth and development was normal in transplants of mammary epithelium from Egfr-/- mice into wild-type (WT) gland free fat pads and in tissue recombinants prepared with WT stroma, irrespective of the source of epithelium (StromaWT/Epi-/-, StromaWT/EpiWT). However, ductal growth and branching was impaired in tissue recombinants prepared with Egfr-/- stroma (Stroma-/-/EpiWT, Stroma-/-/Epi-/-). Thus, for ductal morphogenesis, signaling through the EGFR is required only in the stromal component, the mammary fat pad. These data indicate that the EGFR pathway plays a key role in the stromal-epithelial interactions required for mammary ductal growth and branching morphogenesis. In contrast, signaling through the EGFR is not essential for lobulo-alveolar development. Stimulation of lobulo-alveolar development in the mammary gland grafts by inclusion of a pituitary isograft under the renal capsule as a source of prolactin resulted in normal alveolar development in both Egfr-/- and wild-type transplants. Through the use of tissue recombinants and transplantation, we have gained new insights into the nature of stromal epithelial interactions in the mammary gland, and how they regulate ductal growth and branching morphogenesis. PMID- 9847248 TI - Nuclear beta-catenin is required to specify vegetal cell fates in the sea urchin embryo. AB - Beta-catenin is thought to mediate cell fate specification events by localizing to the nucleus where it modulates gene expression. To ask whether beta-catenin is involved in cell fate specification during sea urchin embryogenesis, we analyzed the distribution of nuclear beta-catenin in both normal and experimentally manipulated embryos. In unperturbed embryos, beta-catenin accumulates in nuclei that include the precursors of the endoderm and mesoderm, suggesting that it plays a role in vegetal specification. Using pharmacological, embryological and molecular approaches, we determined the function of beta-catenin in vegetal development by examining the relationship between the pattern of nuclear beta catenin and the formation of endodermal and mesodermal tissues. Treatment of embryos with LiCl, a known vegetalizing agent, caused both an enhancement in the levels of nuclear beta-catenin and an expansion in the pattern of nuclear beta catenin that coincided with an increase in endoderm and mesoderm. Conversely, overexpression of a sea urchin cadherin blocked the accumulation of nuclear beta catenin and consequently inhibited the formation of endodermal and mesodermal tissues including micromere-derived skeletogenic mesenchyme. In addition, nuclear beta-catenin-deficient micromeres failed to induce a secondary axis when transplanted to the animal pole of uninjected host embryos, indicating that nuclear beta-catenin also plays a role in the production of micromere-derived signals. To examine further the relationship between nuclear beta-catenin in vegetal nuclei and micromere signaling, we performed both transplantations and deletions of micromeres at the 16-cell stage and demonstrated that the accumulation of beta-catenin in vegetal nuclei does not require micromere-derived cues. Moreover, we demonstrate that cell autonomous signals appear to regulate the pattern of nuclear beta-catenin since dissociated blastomeres possessed nuclear beta-catenin in approximately the same proportion as that seen in intact embryos. Together, these data show that the accumulation of beta-catenin in nuclei of vegetal cells is regulated cell autonomously and that this localization is required for the establishment of all vegetal cell fates and the production of micromere-derived signals. PMID- 9847249 TI - Physical and genetic interactions between Alx4 and Cart1. AB - Alx4 and Cart1 are closely related members of the family of transcription factors that contain the paired-type homeodomain. In contrast to other types of homeodomains, the paired-type homeodomain has been shown to mediate high-affinity sequence-specific DNA binding to palindromic elements as either homodimers or as heterodimers with other family members. Alx4 and Cart1 are co-expressed at several sites during development, including the craniofacial mesenchyme, the mesenchymal derivatives of neural crest cells in the first branchial arch and the limb bud mesenchyme. Because of the molecular similarity and overlapping expression pattern, we have analyzed the functional and genetic relationships between Alx4 and Cart1. The two proteins have similar DNA-binding activity in vitro and can form DNA-binding heterodimers; furthermore, they activate transcription of reporter genes that contain high-affinity DNA-binding sites in cell culture in a similar manner. Therefore, at least by these criteria, the two proteins are functionally redundant. Analysis of double mutant animals reveals several genetic interactions. First, mutation of Cart1 exacerbates Alx4-dependent polydactyly in a manner that is dependent on gene dosage. Second, there are complex genetic interactions in the craniofacial region that reveal a role for both genes in the fusion of the nasal cartilages and proper patterning of the mandible, as well as other craniofacial structures. Third, double mutant mice show a split sternum that is not detected in mice with any other genotype. Interpreted in the context of the biochemical characterization, the genetic analysis suggests that Alx4 and Cart1 are indeed functionally redundant, and reveal both unique and redundant functions for these genes in development. PMID- 9847250 TI - Desmosomal localization of beta-catenin in the skin of plakoglobin null-mutant mice. AB - Plakoglobin, a protein belonging to the Armadillo-repeat gene family, is the only component that adherens junctions and desmosomes have in common. Plakoglobin null mutant mouse embryos die because of severe heart defects and may exhibit an additional skin phenotype, depending on the genetic background. Lack of plakoglobin affects the number and structure of desmosomes, resulting in visible defects when cells are subjected to increasing mechanical stress, e.g. when embryonic blood starts circulating or during skin differentiation. By analysing plakoglobin-negative embryonic skin differentiation in more detail, we show here that, in the absence of plakoglobin, its closest homologue, beta-catenin, becomes localized to desmosomes and associated with desmoglein. This substitution may account for the relatively late appearance of the developmental defects seen in plakoglobin null-mutant embryos. beta-catenin cannot, however, fully compensate a lack of plakoglobin. In the absence of plakoglobin, there was reduced cell-cell adhesion, resulting in large intercellular spaces between keratinocytes, subcorneal acantholysis and necrosis in the granular layer of the skin. Electron microscopic analysis documented a reduced number of desmosomes, and those present lacked the inner dense plaque and had fewer keratin filaments anchored. Our analysis underlines the central role of plakoglobin for desmosomal assembly and function during embryogenesis. PMID- 9847251 TI - Regulation of Pax6 expression is conserved between mice and flies. AB - Pax6 plays a key role in visual system development throughout the metazoa and the function of Pax6 is evolutionarily conserved. However, the regulation of Pax6 expression during eye development is largely unknown. We have identified two physically distinct promoters in mouse Pax6, P0 and P1, that direct differential Pax6 expression in the developing eye. P0-initiated transcripts predominate in lens placode and corneal and conjunctival epithelia, whereas P1-initiated transcripts are expressed in lens placode, optic vesicle and CNS, and only weakly in corneal and conjunctival epithelia. To further investigate their tissue specific expression, a series of constructs for each promoter were examined in transgenic mice. We identified three different regulatory regions which direct distinct domains of Pax6 expression in the eye. A regulatory element upstream of the Pax6 P0 promoter is required for expression in a subpopulation of retinal progenitors and in the developing pancreas, while a second regulatory element upstream of the Pax6 P1 promoter is sufficient to direct expression in a subset of post-mitotic, non-terminally differentiated photoreceptors. A third element in Pax6 intron 4, when combined with either the P0 or P1 promoter, accurately directs expression in amacrine cells, ciliary body and iris. These results indicate that the complex expression pattern of Pax6 is differentially regulated by two promoters acting in combination with multiple cis-acting elements. We have also tested whether the regulatory mechanisms that direct Pax6 ocular expression are conserved between mice and flies. Remarkably, when inserted upstream of either the mouse Pax6 P1 or P0 promoter, an eye-enhancer region of the Drosophila eyeless gene, a Pax6 homolog, directs eye- and CNS-specific expression in transgenic mice that accurately reproduces features of endogenous Pax6 expression. These results suggest that in addition to conservation of Pax6 function, the upstream regulation of Pax6 has also been conserved during evolution. PMID- 9847252 TI - Differential patterning of ventral midline cells by axial mesoderm is regulated by BMP7 and chordin. AB - Ventral midline cells in the neural tube have distinct properties at different rostrocaudal levels, apparently in response to differential signalling by axial mesoderm. Floor plate cells are induced by sonic hedgehog (SHH) secreted from the notochord whereas ventral midline cells of the rostral diencephalon (RDVM cells) appear to be induced by the dual actions of SHH and bone morphogenetic protein 7 (BMP7) from prechordal mesoderm. We have examined the cellular and molecular events that govern the program of differentiation of RDVM cells under the influence of the axial mesoderm. By fate mapping, we show that prospective RDVM cells migrate rostrally within the neural plate, passing over rostral notochord before establishing register with prechordal mesoderm at stage 7. Despite the co expression of SHH and BMP7 by rostral notochord, prospective RDVM cells appear to be specified initially as caudal ventral midline neurectodermal cells and to acquire RDVM properties only at stage 7. We provide evidence that the signalling properties of axial mesoderm over this period are regulated by the BMP antagonist, chordin. Chordin is expressed throughout the axial mesoderm as it extends, but is downregulated in prechordal mesoderm coincident with the onset of RDVM cell differentiation. Addition of chordin to conjugate explant cultures of prechordal mesoderm and neural tissue prevents the rostralization of ventral midline cells by prechordal mesoderm. Chordin may thus act to refine the patterning of the ventral midline along the rostrocaudal axis. PMID- 9847253 TI - Expression of chicken fibroblast growth factor homologous factor (FHF)-1 and of differentially spliced isoforms of FHF-2 during development and involvement of FHF-2 in chicken limb development. AB - Members of the fibroblast growth factor (FGF) family have been identified as signaling molecules in a variety of developmental processes, including important roles in limb bud initiation, growth and patterning. This paper reports the cloning and characterization of the chicken orthologues of fibroblast growth factor homologous factors-1 and -2 (cFHF-1/cFGF-12 and cFHF-2/cFGF-13, respectively). We also describe the identification of a novel, conserved isoform of FHF-2 in chickens and mammals. This isoform arises by alternative splicing of the first exon of the FHF-2 gene and is predicted to encode a polypeptide with a distinct amino-terminus. Whole-mount in situ hybridization reveals restricted domains of expression of cFHF-1 and cFHF-2 in the developing neural tube, peripheral sensory ganglia and limb buds, and shows that the two cFHF-2 transcript isoforms are present in non-overlapping spatial distributions in the neural tube and adjacent structures. In the developing limbs, cFHF-1 is confined to the posterior mesoderm in an area that encompasses the zone of polarizing activity and cFHF-2 is confined to the distal anterior mesoderm in a region that largely overlaps the progress zone. Ectopic cFHF-2 expression is induced adjacent to grafts of cells expressing Sonic Hedgehog and the zone of cFHF-2 expression is expanded in talpid2 embryos. In the absence of the apical ectodermal ridge or in wingless or limbless mutant embryos, expression of cFHF-1 and cFHF-2 is lost from the limb bud. A role for cFHF-2 in the patterning and growth of skeletal elements is implied by the observation that engraftment of developing limb buds with QT6 cells expressing a cFHF-2 isoform that is normally expressed in the limb leads to a variety of morphological defects. Finally, we show that a secreted version of cFHF-2 activates the expression of HoxD13, HoxD11, Fgf-4 and BMP-2 ectopically, consistent with cFHF-2 playing a role in anterior-posterior patterning of the limb. PMID- 9847254 TI - Should antileukotriene therapies be used instead of inhaled corticosteroids in asthma? Yes. PMID- 9847255 TI - Should antileukotriene therapies be used instead of inhaled corticosteroids in asthma? No. PMID- 9847256 TI - Protective effect of endotoxin instillation on subsequent bacteria-induced acute lung injury in rats. AB - The phagocytic capability afforded by neutrophil influx into the lungs is essential to ward off invading bacteria. The objective of this study was to evaluate the effect of prior neutrophil recruitment induced by alveolar instillation of endotoxin (LPS, 200 micrograms/kg) 16 h before a pulmonary infection caused by instillation of live Pseudomonas aeruginosa ([PYO]: 1.5 x 10(8) colony-forming units [cfu]/kg) in rats. A first series of experiments showed that lipopolysaccharide (LPS) instillation induced recruitment of alveolar neutrophils that were capable, ex vivo, of elastase exocytosis, reactive oxygen species secretion, and PYO killing. In a second set of experiments, LPS followed by PYO was compared with PYO alone (n = 11 surviving rats in each group). Parameters were studied 24 h after the bacterial challenge. As compared with PYO alone, pretreatment with LPS followed by PYO was associated with decreased mortality (0% versus 54%, p < 0.05), decreased protein leakage into bronchoalveolar lavage (BAL) fluid (1.8 +/- 0.4 versus 13.5 +/- 2.2 mg/ml, p < 0.001), and improved bacterial clearance from BAL (4.0 +/- 1.4 x 10(2) versus 1.2 +/- 0.5 x 10(4) cfu/ml, p < 0.05) and from pulmonary parenchyma (8.5 +/- 6.4 x 10(5) versus 1.9 +/- 0.8 x 10(7) cfu/ml, p < 0.05). We conclude that prior alveolar endotoxin instillation induces local recruitment of functionally active neutrophils, and that this is associated with resistance to subsequent experimental pneumonia. PMID- 9847257 TI - Chronic obstructive pulmonary disease is associated with an increase in urinary levels of isoprostane F2alpha-III, an index of oxidant stress. AB - Oxidative stress has been suggested as a potential mechanism in the pathogenesis of chronic obstructive pulmonary disease (COPD). It has been difficult to address this hypothesis because of the limitations of conventional indices of lipid peroxidation in vivo. F2-isoprostanes (iPs) are prostaglandin isomers formed by free radical dependent peroxidation of arachidonic acid. Urinary iPF2alpha-III is a relatively abundant iPs produced in humans. In the present study, we investigated whether COPD is associated with enhanced oxidative stress by measuring urinary levels of this compound. Urinary excretion of iPF2alpha-III was determined in 38 patients with COPD and 30 sex- and age-matched healthy control subjects. Levels of iPF2alpha-III were significantly higher in patients with COPD (median, 84 pmol/ mmol creatinine; range, 38 to 321) than in healthy controls (median, 35.5 pmol/mmol creatinine; range, 15 to 65) (p < 0.0001). This elevation was independent of age, sex, smoking history, or duration of the disease. An inverse relationship was observed with the level of PaO2 (r = -0.38, p = 0. 019). Aspirin treatment failed to decrease urinary levels of iPF2alpha-III (102 +/- 8 versus 99.2 +/- 7.3 pmol/ mmol creatinine), whereas 11-dehydro TxB2 was significantly reduced (695 +/- 74 versus 95 +/- 10 pmol/mmol creatinine) (p < 0.0001). Elevated levels of iPF2alpha-III (median, 125 pmol/mmol creatinine; range, 110 to 170) in five patients with COPD declined (median, 90 pmol/mmol creatinine; range, 70 to 110) (p < 0.001) as an acute exacerbation in their clinical condition resolved. Increased urinary iPF2alpha-III is consistent with the hypothesis that oxidative stress occurs in COPD. This provides a basis for dose finding and evaluation of antioxidant therapy in the treatment of this disease. PMID- 9847258 TI - In vivo quantitation of epithelial lining fluid in dog lung. AB - We used an original saturation bronchoalveolar lavage (SBAL) technique (Eur. Respir. J. 1995;8[Suppl. 19]398S) to quantitate lung epithelial lining fluid volume (VELF) in dogs in two separate experiments: control and after oleic-acid induced injury. We confirmed the hypothesis that 99mTc-DTPA, infused at constant plasma activity, reaches equilibrium with epithelial lining fluid after 90 min. We performed eight sequential lavages 215 min after beginning the infusion of 99mTc-DTPA. 99mTc-DTPA activity (Qn) in the lavage fluid increased linearly with time, suggesting transport from the plasma into the alveoli during lavage. We extrapolated Qn to time zero (Q0), when 99mTc-DTPA was not affected by lavage. VELF was calculated from: VELF = Q0/Cp, (Cp: 99mTc-DTPA mean plasma activity). 125I-albumin was used as a nondiffusible alveolar indicator to measure the fluid volume present in the lavaged segment (Vt,n). Vt,n plateaud for n >= 4. VELF/Vt,n(n = 5,8) was 1.7 +/- 0.4 and 25.0 +/- 4.4% (p < 0.05) in control and injury experiments, respectively. SBAL allowed reliable measurements of VELF and detection of alveolar edema fluid in the injured lung. PMID- 9847259 TI - Respiratory symptoms and lung function in aborigines from tropical Western Australia. AB - To estimate the prevalence of respiratory symptoms, bronchial hyperresponsiveness, smoking, and atopy in a population of Australians of Aboriginal descent (AAD), to determine the association of these and other factors with lung function, and to compare levels of lung function of AAD with Australians of European descent (AED) according to age and height, and to explore reasons for their differences, we conducted a study of 96 male (41 of whom were under 18 yr of age) and 111 female (48 of whom were under 18 yr of age) AAD living in a single remote tropical community in 1993. This population provided data on age, height, and lung function. A modified British Medical Research Council (MRC) questionnaire on respiratory symptoms and smoking was administered. FEV1, FVC, height, age, and bronchial responsiveness to inhaled methacholine were measured. Atopic status was assessed by skin prick tests for eight common allergens. Age- and sex-adjusted lung function was similar to that of other AAD groups and lower than in AED. For children, lung function increased less with increasing height in AAD than in AED. Lung function was reduced in adult AAD as compared with adult AED, although it was not possible to determine statistically whether lung function started to decline at an earlier age or declined faster with increasing age in AAD. A history of asthma, smoking, dyspnea, cough, or sputum production; atopic status; and increased bronchial responsiveness were all associated with lower levels of lung function. Differences in lung function between AAD and AED appear to be determined by characteristics that may be inherited, as well as by adverse external influences. PMID- 9847260 TI - Active detection of chronic obstructive pulmonary disease and asthma in the general population. Results and economic consequences of the DIMCA program. AB - The aim of this prospective study was to detect subjects in the general population with objective signs of chronic obstructive pulmonary disease (COPD) or asthma at an early stage. This was done by means of a two-stage protocol involving screening and a subsequent 2-yr monitoring of all subjects with positive results of screening. The study was done in 10 general practices located in the eastern part of the Netherlands. A random sample was taken from the general population aged 25 to 70 yr. All known COPD and asthma patients were excluded. A total of 1,749 subjects met the inclusion criteria: 1,155 subjects (66%) agreed to participate in the screening stage of the study. A total of 604 subjects (52.3%) showed symptoms or objective signs of COPD or asthma during the screening and were considered "positive." Of those with positive screening results, 384 subjects (64%) agreed to participate in the second, 2-yr monitoring stage of the study. The costs involved in detection were calculated for three different scenarios, as follows: (1) The detection of subjects with persistently decreased lung function or an increased level of bronchial hyperresponsiveness (BHR) during 6 mo of monitoring; (2) Scenario 1 plus the detection of subjects with a rapid decline in lung function with signs of BHR during 12 mo of monitoring; (3) Scenario 2 plus the detection of subjects with a moderate increase in the decline in lung function or signs of BHR during 24 mo of monitoring. The costs of lung function assessments, organization, transportation, and patient time were included. The costs were converted to United States dollars on the basis of purchasing power (1 United States dollar = 2.08 Netherlands guilders). During the second stage, 252 subjects were detected with objective signs of COPD or asthma at an early stage. Smoking status as a screening criterion was neither sensitive nor specific. Because there was no evidence of biased recruitment or selection during the program, the proportions of subjects found to have objective signs of COPD or asthma at an early stage could be extrapolated to the general population. Of the general population, 7.7% showed persistently reduced lung function or increased BHR. Another 12.5 % of the general population showed a rapid decline in lung function (> 80 ml/yr) in combination with signs of BHR, and a further 19.4% of the general population showed mild objective signs of COPD or asthma. The average costs per detected case varied from US$953 (Scenario 1) to US$469 (Scenario 3). In conclusion, detection of COPD or asthma at an early stage by means of a two-stage protocol was feasible at relatively little expense in comparison with other mass screening programs. Persistently decreased lung function or a rapid decline in lung function (Scenario 2) was observed in approximately 20% of the general adult population. PMID- 9847261 TI - Linkage of circulating eosinophils to markers on chromosome 5q. AB - Although peripheral blood eosinophilia is strongly associated with the risk of developing asthma, genetic determinants of eosinophilia have not been extensively studied. We used sib-pair analysis to assess linkage of circulating eosinophils (as a percent of total white blood cells [WBC]) to nine markers located in chromosome 5q31-33. The study was divided into two phases. Of 246 sib pairs available for the first phase, 35 were classified as low concordant (LC) (both sibs had <= 2% circulating eosinophils), 18 were defined as high concordant (HC) (both sibs had 5% or more circulating eosinophils), and 26 were defined as discordant (one sib had <= 2% and the other sib had 5% or more circulating eosinophils). Significant evidence for linkage among low concordant sib pairs was found for several markers in the region under study, with a peak for marker D5S500 (proportion of alleles shared identical by descent [ibd] = 0.68 +/- 0.05 [mean +/- SE], p = 0.0004). A cross-validating study was done in which an additional 19 sib pairs that were low concordant for circulating eosinophils were studied. Evidence for linkage was also observed in this subset. Results were independent of current wheezing, total serum IgE levels, and other potential confounders. A multipoint analysis done for all low-concordant sib pairs available showed that the maximal logarithm of the odds favoring genetic linkage (LOD) score (2.4, p = 0.0004) was observed in correspondence with marker D5S658. We conclude that a locus or loci may be present in chromosome 5q31-33 that controls for circulating eosinophils as a proportion of total WBC. PMID- 9847262 TI - Applicability of the single-breath carbon monoxide diffusing capacity in a Norwegian Community Study. AB - The test of single-breath diffusing capacity for carbon monoxide (DLCO) has been widely used in population surveys. However, little is known about the effect of meeting or failing to meet the criteria for acceptability of this test. The American Thoracic Society (ATS) recommends a breathholding time of 9 to 11 s, two measurements within +/- 10% or 3 ml CO(STPD)/min/mm Hg of the average DLCO, and an inspiratory vital capacity (IVC) of at least 90% of the largest previously measured forced vital capacity (FVC) as criteria for this test. The objective of the present study was to examine the extent to which these criteria were met in a community study. To do this, a random sample of 3,740 persons, aged 15 to 70 yr, of the general population of the city of Bergen and 11 surrounding municipalities on the southwest coast of Norway were enrolled in a two-phase cross-sectional study. In the second phase, a stratified sample (n = 1,512) of the respondents to the postal questionnaire used for recruitment for the study (n = 3,370) were invited to a clinical and respiratory physiologic examination that included the DLCO test. The attendance rate was 84% (1,275 of 1,512). In the examination, all subjects were able to maintain a breathholding time of 9 to 11 s, and 98% had two DLCO values within +/- 10% or 3 ml CO(STPD)/min/mm Hg of the average DLCO. The criterion of an IVC of at least 90% of FVC in the two tests was met by 68% of the subjects. Younger age was an independent predictor of failure to meet the required criteria. Thus, only two-thirds of the participants fulfilled all of the ATS criteria for the DLCO test, the main reason for failure being an IVC of less than 90% FVC. This should not necessarily lead to the exclusion from further analysis of those failing to meet this criterion. PMID- 9847263 TI - Response to cooling temperature in infants born at an altitude of 4,330 meters. AB - The metabolic response to reduction in ambient temperature was studied in healthy, full-term, 1-d-old infants in Lima (50 m altitude, n = 20) and Cerro de Pasco (4,330 m, barometric pressure approximately 450 mm Hg, n = 20), Peru. Oxygen consumption (V O2) and carbon dioxide production (V CO2) were measured with an open-flow system as each infant rested quietly in a cylindrical humicrib, at wall temperatures of 35 degrees C (warm) and 26 degrees C (cool). The infants were exposed for 20 min to both temperatures, with the higher temperature followed by the lower, and oxygen consumption (V O2) and carbon dioxide production (V CO2) were measured over the last 8 min of each exposure. Average birth weight in Cerro de Pasco (2,933 +/- 77 g [mean +/- SE]) was less than in Lima (3,457 +/- 73 g). In warm conditions, infants born at high altitude had slightly yet significantly lower body and skin temperatures than did those born at low altitude, with similar values of V O2 and heart rate (HR). Neither body nor skin temperature changed in either group during cooling. At low altitude, cooling increased V O2 ( approximately 34%), whereas no significant increase occurred in the high-altitude group. A similar response occurred for HR. Among several possibilities, the most likely interpretation of the results would be that of a decreased thermogenic capacity in the high-altitude infants because of the correspondingly lower oxygen availability during gestation. PMID- 9847265 TI - Continuous recordings of mixed venous oxygen saturation during weaning from mechanical ventilation and the ramifications thereof. AB - To define the importance of hemodynamic performance and global tissue oxygenation in determining weaning outcome, we recorded mixed venous oxygen saturation (SvO2) continuously in eight ventilator-supported patients who failed a trial of spontaneous breathing and 11 patients who tolerated a trial and were successfully extubated. Immediately before the weaning trial, SvO2 was not statistically different in the two groups (p = 0.28). On discontinuation of the ventilator, SvO2 fell progressively in the failure group (p < 0.01), whereas it did not change in the success group. During the trial of spontaneous breathing, O2 demand was similar in the two groups, but it differed in the manner with which it was met. The success group demonstrated an increase in cardiac index (p < 0.05) and O2 transport (p < 0.02). The failure group did not increase O2 transport, partly because of elevations in right- and left-ventricular afterload, but, instead, increased O2 extraction ratio (p < 0.02) with a consequent fall in SvO2. In turn, the low SvO2 combined with greater venous admixture (p < 0.0006) led to rapid arterial desaturation (p < 0.006) and a relative decrease in O2 being supplied to the tissues. In conclusion, ventilator-supported patients who failed a trial of spontaneous breathing developed a progressive decrease in SvO2 caused by the combination of a relative decrease in convective O2 transport and an increase in O2 extraction by the tissues. PMID- 9847264 TI - Hypoxia enhances cellular proliferation and inositol 1,4, 5-triphosphate generation in fibroblasts from bovine pulmonary artery but not from mesenteric artery. AB - When pulmonary hypertension occurs in the face of hypoxia there is remodeling of all three layers of the pulmonary vessels, but in particular, there is an increase in number of adventitial fibroblasts. Hypoxia causes vasoconstriction in the pulmonary circulation and vasodilation in the systemic circulation. We hypothesized that there are fundamental differences in oxygen sensing and cell signaling between systemic and pulmonary artery cells in response to hypoxia. Here, we determined the effect of hypoxia either alone or in combination with known growth factors such as serum, endothelin-1 (ET-1), and platelet-derived growth factor (PDGF) on the proliferative responses of bovine pulmonary artery and mesenteric artery fibroblasts. Fibroblasts were obtained from primary cultures. Growth was assessed by [3H]thymidine incorporation. Inositol 1,4,5 triphosphate (IP3) generation was measured using a competitive binding assay. Hypoxia alone increased proliferation of pulmonary artery fibroblasts (611 +/- 24%), but not in those from the mesentery. Furthermore, hypoxia had the effect of increasing the replicative response of pulmonary fibroblasts to serum and PDGF, but no change was observed in the mesenteric cells. ET-1 had no effect on growth of either cell type. PDGF gave rise to a significant elevation in IP3 production under hypoxic conditions in the pulmonary artery cells (234%), but not in the mesenteric cells. ET-1 caused no change in IP3 production in any cell type. These data suggest that hypoxia sensitizes pulmonary artery fibroblasts to the proliferative effect of mitogens through a pathway that is not present, or is present but repressed, in the mesenteric cells. PMID- 9847266 TI - Intravenous almitrine combined with inhaled nitric oxide for acute respiratory distress syndrome. The NO Almitrine Study Group. AB - Inhaled nitric oxide (iNO), a selective pulmonary vasodilator and intravenously administered almitrine, a selective pulmonary vasoconstrictor, have been shown to increase PaO2 in patients with acute respiratory distress syndrome (ARDS). This prospective study was undertaken to assess the cardiopulmonary effects of combining both drugs. In 48 consecutive patients with early ARDS, cardiorespiratory parameters were measured at control, after iNO 5 ppm, after almitrine 4 micrograms. kg-1. min-1, and after the combination of both drugs. In 30 patients, dose response to 2, 4, and 16 micrograms. kg-1. min-1 of almitrine with and without NO was determined. Almitrine and lactate plasma concentrations were measured in 17 patients. Using pure O2, PaO2 increased by 75 +/- 8 mm Hg after iNO, by 101 +/- 12 mm Hg after almitrine 4 micrograms. kg-1. min-1, and by 175 +/- 18 mm Hg after almitrine combined with iNO (p < 0.001). In 63% of the patients, PaO2 increased by more than 100% with the combination of both drugs. Mean pulmonary artery pressure (Ppa) increased by 1.4 +/- 0.2 mm Hg with almitrine 4 micrograms/kg/ min (p < 0.001) and decreased by 3.4 +/- 0.4 mm Hg with iNO and by 1.5 +/- 0.3 mm Hg with the combination (p < 0.001). The maximum increase in PaO2 was obtained at almitrine concentrations <= 4 micrograms. kg-1. min-1, whereas almitrine increased Ppa dose-dependently. Almitrine plasma concentrations also increased dose-dependently and returned to values close to zero after 12 h. In many patients with early ARDS, the combination of iNO 5 ppm and almitrine 4 micrograms. kg-1. min-1 dramatically increases PaO2 without apparent deleterious effect allowing a rapid reduction in inspired fraction of O2. The long-term consequences of this immediate beneficial effect remain to be determined. PMID- 9847267 TI - Differentiating obstructive and central sleep respiratory events through pulse transit time. AB - Noninvasive alternatives to esophageal pressure (Pes) are needed to evaluate respiratory effort during sleep. Pulse transit time (PTT) is the time taken for pulse pressure to travel from the aortic valve to the periphery. PTT has been shown to be inversely correlated with blood pressure, and can reveal acute changes generated by high pleural pressure swings during pulsus paradoxus. A close relationship has been demonstrated between the increase in Pes and a progressive rise in the amplitude of PTT oscillations. The aim of the present study was to assess the accuracy of PTT for the classification of sleep respiratory events as central or obstructive. Respiratory events occurring during sleep were randomly chosen from 13 unselected male patients (mean apnea-hypopnea index [AHI] = 25.1 per hour of sleep; age = 47.3 yr, body mass index [BMI] = 27.1 kg/m2). Two observers experienced in polysomnography classified 177 events on the basis of the "gold standard method": the measurement of Pes. For 167 events about which the observers agreed, the PTT signal was analyzed visually and independently by the two observers blinded to Pes, in order to reclassify the same sleep respiratory events. The two observers were in agreement for 94.6% of the events scored visually on PTT recordings. We evaluated sensitivity (Se) (Observer 1: 94%, Observer 2: 91%), specificity (Sp) (97% and 95%, respectively), negative predictive value (NPV) (95% and 92%, respectively), and positive predictive value (PPV) (96% and 94%, respectively), of PTT with Pes as the reference. Misclassifications of respiratory episodes were usually due to artifacts or baseline variations of the PTT signal (57%), and occurred during rapid eye movement (REM) sleep (42.8%). PTT has shown a high sensitivity and specificity in differentiating obstructive and central respiratory events, and may become the reference noninvasive tool for this purpose. PMID- 9847268 TI - Effects of endogenous superoxide anion and nitric oxide on cholinergic constriction of normal and hyperreactive guinea pig airways. AB - In a guinea pig model of allergic asthma, we have recently established that a deficiency of nitric oxide (NO) contributes to the increased ex vivo responsiveness of isolated perfused tracheae to methacholine after the early asthmatic reaction at 6 h after inhalational challenge of the animals with ovalbumin aerosol. Because this deficiency could be caused by a reaction of NO with enhanced levels of inflammation-induced superoxide anion (O-2), we examined the effect of endogenous O-2 on the regulation of methacholine-induced constriction by NO of intact perfused tracheal tube preparations from unchallenged (control) guinea pigs and from animals 6 h after ovalbumin challenge. In the presence of the NO synthase (NOS) inhibitor N omega-nitro-L arginine methyl ester (L-NAME; 100 microM), tracheae obtained from unchallenged guinea pigs showed a 1.7-fold increase in the maximal response to intraluminally applied methacholine (p < 0.05). By contrast, the maximal airway response to methacholine was significantly decreased in the presence of the O-2 scavenger superoxide dismutase (SOD; 100 U/ml), by approximately 45% (p < 0.01). The SOD induced decrease in responsiveness to methacholine was reversed by L-NAME. Tracheal preparations obtained at 6 h after allergen challenge showed a 1. 8-fold increased responsiveness to intraluminally applied methacholine compared with controls (p < 0.001), which was not further enhanced in the presence of L-NAME. SOD had neither an effect on the increased responsiveness nor did it restore the potentiating effect of L-NAME. These results indicate that (1) in normoreactive tracheal preparations, the regulatory role of NO is partially counteracted by endogenous O-2, and ( 2) the deficiency of NO in hyperreactive tracheae obtained at 6 h after ovalbumin challenge is not caused by its reaction with O-2, but rather to decreased cNOS activity. De Boer J, Pouw FMH, Zaagsma J, Meurs H. Effects of endogenous superoxide anion and nitric oxide on cholinergic constriction of normal and hyperreactive guinea pig airways. PMID- 9847269 TI - Instability of tuberculin and Candida skin test reactivity in HIV-infected Ugandans. The Uganda-Case Western Reserve University Research Collaboration. AB - Anergy testing has been used as an adjunct to tuberculin testing for assessing M. tuberculosis (MTB) infection and indications for isoniazid preventive therapy in HIV-infected persons. We examined factors associated with the stability of skin test responses to purified protein derivative (PPD) and candida antigens in a cohort of HIV-infected adults followed prospectively in a tuberculosis preventive therapy trial in Uganda. PPD-positive and anergic subjects in the placebo arms of the preventive therapy study underwent repeat skin testing and immunologic testing including measurement of MTB culture filtrate (CF)-stimulated interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) levels in whole blood culture supernatants. Anergy was present in 27% of 4,058 HIV-infected subjects screened for the tuberculosis preventive therapy trial compared with 10% of 682 HIV-non-infected persons. On follow-up testing of enrolled subjects, 42% of 139 initially anergic subjects were no longer anergic; two thirds of these had PPD reactions >= 5 mm. Stability of anergy was associated with intercurrent opportunistic infections and AIDS-associated dermatitis at baseline. Thirty-five percent of 313 subjects with an initial positive PPD had a negative PPD test at follow-up, 26% of whom had a positive candida skin test at the same time as the negative PPD test. Baseline MTBCF-stimulated IFN-gamma levels were significantly higher among PPD-positive subjects who remained PPD-positive than in those who were falsely negative. We conclude first that anergy is unstable and second that anergy testing is unreliable in identifying HIV-infected adults who are not infected with MTB and should not be used routinely for this purpose in assessing indications for isoniazid preventive therapy. PMID- 9847270 TI - Differences in contributing factors to tuberculosis incidence in U.S. -born and foreign-born persons. AB - To determine the factors contributing to tuberculosis incidence in the U.S.-born and foreign-born populations in San Francisco, California, and to assess the effectiveness of tuberculosis control efforts in these populations, we performed a population-based molecular epidemiologic study using 367 patients with strains of Mycobacterium tuberculosis recently introduced into the city. IS6110-based and PGRS-based restriction fragment length polymorphism (RFLP) analyses were performed on M. tuberculosis isolates. Patients whose isolates had identical RFLP patterns were considered a cluster. Review of public health and medical records, plus patient interviews, were used to determine the likelihood of transmission between clustered patients. None of the 252 foreign-born cases was recently infected (within 2 yr) in the city. Nineteen (17%) of 115 U. S.-born cases occurred after recent infection in the city; only two were infected by a foreign born patient. Disease from recent infection in the city involved either a source or a secondary case with human immunodeficiency virus (HIV) infection, homelessness, or drug abuse. Failure to identify contacts accounted for the majority of secondary cases. In San Francisco, disease from recent transmission of M. tuberculosis has been virtually eliminated from the foreign-born but not from the U.S.-born population. An intensification of contact tracing and screening activities among HIV-infected, homeless, and drug-abusing persons is needed to further control tuberculosis in the U.S.-born population. Elimination of tuberculosis in both the foreign-born and the U.S. -born populations will require widespread use of preventive therapy. PMID- 9847271 TI - Increased prevalence of gastroesophageal reflux in patients with idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is a progressive, fatal interstitial lung disease (ILD) of unknown etiology. Introduction of acid into the respiratory tree can produce pulmonary fibrosis. Gastroesophageal reflux (GER) has previously been associated with several other respiratory conditions, including pneumonia, bronchitis, and asthma. To investigate prospectively the possible association of GER and IPF, 17 consecutive patients with biopsy-proven IPF and eight control patients with ILD other than IPF underwent dual-channel, ambulatory esophageal pH monitoring. Sixteen of 17 patients with IPF had abnormal distal and/or proximal esophageal acid exposure compared with four of eight control patients (p = 0.02). In the patients with IPF, mean percent distal total (13.6 versus 3.34, p = 0.006), distal upright (12.4 versus 5.1, p = 0.04), distal supine (14.7 versus 0.88, p = 0.02), and proximal supine (7.48 versus 0.24, p = 0.04) esophageal acid exposure times were significantly greater than those in control patients. Only four patients with IPF (25%) with increased acid exposure had typical reflux symptoms such as heartburn or regurgitation. Patients with IPF have a high prevalence of increased esophageal acid exposure, usually without typical GER symptoms. GER in these patients tends to occur at night and extend into the proximal esophagus. Acid reflux may be a contributing factor in the pathogenesis of IPF. PMID- 9847272 TI - Anti-inflammatory agents and allergen-induced beta2-receptor dysfunction in isolated human bronchi. AB - Antigen challenge causes beta2-adrenoceptor dysfunction in sensitized human bronchi (Am. J. Respir. Crit. Care Med. 1997;155:1230-1234). This study investigated whether the dysfunction can be prevented by anti-inflammatory agents. Human bronchial rings (2 to 4 mm) from surgery were passively sensitized to house dust mite and challenged (1) with allergen only, (2) with allergen plus indomethacin (10(-)5 M), (3) with allergen plus nedocromil sodium (10(-)7 M to 10(-)5 M), (4) with allergen plus the H1-receptor antagonist cetirizine (10(-)7 M to 10(-)5 M), and (5) with allergen plus the peptido-leukotriene receptor antagonist iralukast (10(-)7 M to 10(-)5 M). Rings were first contracted with 10( )6 M carbachol and then relaxed with salbutamol (10(-)9 M to 10(-)4 M). The concentration-relaxation curve to salbutamol was shifted significantly to the right in the rings challenged with allergen only compared with control rings. In the rings challenged with allergen plus nedocromil sodium (10(-)6 M and 10(-)5 M) or iralukast (10(-)6 M and 10(-)5 M) the concentration-relaxation curves to salbutamol were significantly shifted to the left compared with rings challenged in saline alone, suggesting a protective effect against beta2-adrenoceptor dysfunction. Neither allergen plus cetirizine nor allergen plus indomethacin shifted significantly the concentration-relaxation curves to salbutamol compared with rings challenged in saline alone. We conclude that the release of peptido leukotrienes may play a significant role in causing the allergen-induced beta2 receptor dysfunction in passively sensitized human bronchi. PMID- 9847273 TI - Assessment of environmental asbestos exposure in Turkey by bronchoalveolar lavage. AB - Environmental or domestic exposure to asbestos fibers originating from local soil is responsible for a high incidence of diseases in large rural areas of Turkey. Bronchoalveolar lavage fluids (BALF) were obtained for 65 Turkish subjects originating from these areas and for 42 Turkish controls. Asbestos bodies (ABs) and uncovered fibers (UFs) were quantified by phase contrast light microscopy. Total fiber burden was determined by transmission electron microscopy. The main asbestos types disclosed were tremolite and to a lesser extent chrysotile. AB and fiber concentrations were higher in environmentally exposed subjects (geometric mean [geometric standard deviation]: 5.20 [6.22] AB/ml, 444 [11.6] tremolite fibers/ml) than in control subjects (0.22 [1.45] AB/ml, 12.0 [15.4] tremolite fibers/ml) (p < 0.001). In subjects environmentally exposed in Turkey, AB burdens on tremolite were in the same range as those on commercial amphiboles in subjects occupationally exposed in Belgium. In Turkish subjects, values above either 1 AB/ml, 3 uncovered fiber/ml in light microscopy, or 300 fibers/ml in electron microscopy indicated usually an abnormal alveolar retention reflecting a significant cumulative exposure from environmental or domestic origin. These observations are probably valid for other areas in the world where diseases associated with environmental exposure to soil- derived asbestos fibers occur and for immigrants originating from these areas. PMID- 9847274 TI - Linkage of chromosome 5q and 11q gene markers to asthma-associated quantitative traits in Australian children. AB - Asthma is a genetically complex disease, and the investigation of putative linkages to candidate loci in independent populations is an important part of the gene discovery process. This study investigated the linkage of microsatellite markers in the 5q and 11q regions to asthma-associated quantitative traits in 121 Australian Caucasian nuclear families. The families were recruited on the basis of a child proband: a cohort of 95 randomly recruited families of unselected probands (n = 442 subjects) and a cohort of 26 families of probands selected on the basis of severe symptomatic asthma (n = 134 subjects). The quantitative traits assessed included serum levels of total IgE and specific IgE to house dust mite and mixed grass, blood eosinophil counts, and the dose-response slope (DRS) of FEV1 to histamine provocation. Multipoint linkage analysis using Haseman Elston sib-pair methods provided evidence of significant linkage between the chromosome 5q markers and loge total serum IgE levels, specific serum IgE levels, and loge blood eosinophil counts. The chromosome 11q markers showed evidence of significant linkage to specific serum IgE levels. Neither region demonstrated significant linkage to the loge DRS to histamine. Phenotypes were residualized for age and sex. These data are consistent with the existence of loci regulating asthma-associated quantitative traits in both the 5q31-33 and 11q13 chromosomal regions. PMID- 9847275 TI - Tidal volume reduction for prevention of ventilator-induced lung injury in acute respiratory distress syndrome. The Multicenter Trail Group on Tidal Volume reduction in ARDS. AB - Because animal studies have demonstrated that mechanical ventilation at high volume and pressure can be deleterious to the lungs, limitation of airway pressure, allowing hypercapnia if necessary, is already used for ventilation of acute respiratory distress syndrome (ARDS). Whether a systematic and more drastic reduction is necessary is debatable. A multicenter randomized study was undertaken to compare a strategy aimed at limiting the end-inspiratory plateau pressure to 25 cm H2O, using tidal volume (VT) below 10 ml/kg of body weight, versus a more conventional ventilatory approach (with regard to current practice) using VT at 10 ml/kg or above and close to normal PaCO2. Both arms used a similar level of positive end-expiratory pressure. A total of 116 patients with ARDS and no organ failure other than the lung were enrolled over 32 mo in 25 centers. The two groups were similar at inclusion. Patients in the two arms were ventilated with different VT (7.1 +/- 1.3 versus 10.3 +/- 1.7 ml/kg at Day 1, p < 0.001) and plateau pressures (25.7 +/- 5. 0 versus 31.7 +/- 6.6 cm H2O at Day 1, p < 0.001), resulting in different PaCO2 (59.5 +/- 15.0 versus 41.3 +/- 7.6 mm Hg, p < 0.001) and pH (7.28 +/- 0.09 versus 7.4 +/- 0.09, p < 0.001), but a similar level of oxygenation. The new approach did not reduce mortality at Day 60 (46.6% versus 37.9% in control subjects, p = 0.38), the duration of mechanical ventilation (23.1 +/- 20.2 versus 21.4 +/- 16. 3 d, p = 0.85), the incidence of pneumothorax (14% versus 12%, p = 0. 78), or the secondary occurrence of multiple organ failure (41% versus 41%, p = 1). We conclude that no benefit could be observed with reduced VT titrated to reach plateau pressures around 25 cm H2O compared with a more conventional approach in which normocapnia was achieved with plateau pressures already below 35 cm H2O. PMID- 9847276 TI - Epidemiology of ventilator-acquired pneumonia based on protected bronchoscopic sampling. AB - We performed a prospective observational cohort study of the epidemiology and etiology of nosocomial pneumonia in 358 medical ICU patients in two university affiliated hospitals. Protected bronchoscopic techniques (protected specimen brush and bronchoalveolar lavage) were used for diagnosis to minimize misclassification. Risk factors for ventilator-associated pneumonia were identified using multiple logistic regression analysis. Twenty-eight cases of pneumonia occurred in 358 patients for a cumulative incidence of 7.8% and incidence rates of 12.5 cases per 1, 000 patient days and 20.5 cases per 1,000 ventilator days. Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Hemophilus species made up 65% of isolates from the lower respiratory tract, whereas only 12.5% of isolates were enteric gram-negative bacilli. Daily surveillance cultures of the nares, oropharynx, trachea, and stomach demonstrated that tracheal colonization preceded ventilator-associated pneumonia in 93.5%, whereas gastric colonization preceded tracheal colonization for only four of 31 (13%) eventual pathogens. By multiple logistic regression, independent risk factors for ventilator- associated pneumonia were admission serum albumin <= 2.2 g/dl (odds ratio [OR] 5.9; 95% confidence interval [CI] 2.0 17.6; p = 0.0013), maximum positive end-expiratory pressure >= 7.5 cm H2O (OR, 4.6; 95% CI, 1.4 to 15.1; p = 0.012), absence of antibiotic therapy (OR, 6.7; 95% CI, 1.8 to 25.3; p = 0.0054), colonization of the upper respiratory tract by respiratory gram-negative bacilli (OR, 3.4; 95% CI, 1.1 to 10.1; p = 0.028), pack years of smoking (OR, 2.3 for 50 pack-years; 95% CI, 1. 2 to 4.2; p = 0.012), and duration of mechanical ventilation (OR, 3. 4 for 14 d; 95% CI, 1.5 to 7.8; p = 0.0044). Several of these risk factors for ventilator-associated pneumonia appear amenable to intervention. PMID- 9847278 TI - Clinical characteristics, respiratory functional parameters, and outcome of a two hour T-piece trial in patients weaning from mechanical ventilation. AB - The discrepancy in results from different studies regarding outcome of weaning from mechanical ventilation may be due to several factors such as the differences in patient populations and weaning indexes used. In order to analyze the clinical characteristics and weaning indexes in patients undergoing a 2-h T-piece weaning trial and the relationship between the etiology of acute respiratory failure (ARF) and the outcome of this weaning trial, we prospectively studied 217 patients receiving mechanical ventilation who met standard weaning criteria. Successful weaning occurred in 57.6% (125 of 217) of patients: 13 of 33 (39.4%) patients with chronic obstructive pulmonary disease (COPD), 27 of 46 (58.7%) neurologic patients, and 85 of 138 (61.6%) patients with ARF. Ventilatory support was reinstituted in 31.8% (69 of 217) patients: 20 of 33 (60.6%) of patients with COPD, four of 46 (8.7%) neurologic patients, and 45 of 138 (32.6%) patients with ARF (p < 0.001). Reintubation was required in 23 of 148 (15.5%) patients: 15 of 42 (35.7%) neurologic patients, and eight of 93 (8.6%) patients with ARF, whereas no patient with COPD was reintubated (p < 0.001). Using a discriminant analysis, the following variables were selected as the best predictors of outcome: (1) in the whole population, days of mechanical ventilation before weaning trial (DMV), frequency-to-tidal volume ratio (f/VT), maximal inspiratory pressure (MIP), airway occlusion pressure (P0.1), maximal expiratory pressure (MEP), and vital capacity (VC); (2) in patients with ARF, DMV, P0.1/MIP, MIP, f/VT, and age; (3) in patients with COPD, f/VT, P0.1, P0.1/MIP, MIP, age, and DMV; (4) in neurologic patients, MIP, MEP, and f/VT.P0.1. Using these predictors, 74.6% of the whole population, 76.1% of patients with ARF, 93.9% of patients with COPD, and 73.9% of neurologic patients were accurately classified as weaning successes or failures. The highest rate of reintubation occurred in neurologic patients. In this group, the ability to cough and clear respiratory secretions, objectively reflected by MEP, may help in clinical decision-making. PMID- 9847277 TI - Peak expiratory flow variability, bronchial responsiveness, and susceptibility to ambient air pollution in adults. AB - Bronchial hyperresponsiveness (BHR) and peak expiratory flow (PEF) variability are associated expressions of airway lability, yet probably reflect different underlying pathophysiologic mechanisms. We investigated whether both measures can be used interchangeably to identify subjects who are susceptible to ambient air pollution. Data on BHR (>= 20% fall in FEV1), PEF variability (ampl%mean PEF > 5% on any day during an 8-d period with low air pollution levels) and diary data on upper and lower respiratory symptoms, cough, and phlegm were collected in 189 subjects (48-73 yr). The acute effects (lag0) of particulate matter with a diameter less than 10 micrometers (PM10), black smoke, SO2 and NO2 on the prevalence of symptoms were estimated with logistic regression. In subjects with airway lability, both when expressed as PEF variability (69%) and BHR (28%), the prevalence of symptoms increased significantly with increasing levels of air pollution, especially in those with the greater PEF variability (n = 55, 29%). We found no such consistent positive associations in adults without airway lability. PEF variability, and to a smaller extent BHR, can be used to identify adults who are susceptible to air pollution. Though odds ratios were rather low (ranging from 1.13 to 1.41), the impact on public health can be substantial because it applies to large populations. PMID- 9847280 TI - Mycobacterium tuberculosis infection after travel to or contact with visitors from countries with a high prevalence of tuberculosis. AB - To assess whether there is increased risk of tuberculous infection in children who traveled to or had a household visitor from a country having a high prevalence of tuberculosis, we conducted a case-control study. Children younger than 6 yr of age who had a tuberculin skin test read at public health clinics in areas of California that have a high prevalence of tuberculosis were enrolled. Of the 953 children who had a skin test read, 72 (7.6%) had a positive reaction. By multiple logistic regression analysis, children who had traveled in the 12 mo before the skin test were 3.9 times more likely to have a positive skin test than were children who had not traveled (95% confidence interval [CI], 1.9 to 7.9). Among children born in the United States, those who had traveled were 4.7 times more likely to have a positive skin test (95% CI, 2.0 to 11.2). Children who had a household visitor from a country having a high prevalence of tuberculosis were 2.4 times more likely to have a positive skin test than were those who did not have a visitor (95% CI, 1.0 to 5.5). These data indicate that travel to a country that has a high prevalence of tuberculosis or having a visitor from such countries increase the risk of tuberculous infection among young children. Physicians and tuberculosis control programs should incorporate questions about travel and visitors into their evaluations, and children with a history of extended travel should have a tuberculin skin test. PMID- 9847279 TI - CD4+ T cells can induce airway hyperresponsiveness to allergen challenge in the brown norway rat. AB - Airway hyperresponsiveness to inhalational challenge with methacholine (MCh) develops by 32 h after allergen challenge of actively sensitized BN rats. To test the hypothesis that CD4+ T cells mediate allergen-induced hyperresponsiveness independent of IgE-mediated mechanisms, we administered CD4+ T cells, CD8+ T cells, and a mixture of CD4+ and CD8+ T cells (total T cells) isolated from the cervical lymph nodes of rats sensitized with ovalbumin (OA) to naive BN rats that underwent aerosol challenge with either OA or bovine serum albumin (BSA) 2 d later. Responsiveness to MCh was measured 2 d before transfer of T cells and 32 h after challenge with OA or BSA. Airway responsiveness increased significantly in recipients of CD4+ T cells after OA challenge, but not in any other of the treatment groups. Analysis of bronchoalveolar lavage (BAL) cells for major basic protein expression by immunostaining showed eosinophilia in OA-challenged CD4+ and total T-cell recipients. Cells retrieved by bronchoalveolar lavage showed increased expression of IL-5 mRNA (in situ hybridization) in CD4+ T cell recipients after OA challenge compared with other groups. Interferon-gamma mRNA was expressed to the greatest extent in CD8+ recipients, but it was elevated in both OA- and BSA-challenged animals. We conclude that CD4+ T cells can induce airway hyperresponsiveness after inhalational challenge with allergen and this is associated with IL-5 production and eosinophilia. CD8+ T cells may have a negative regulatory effect on responsiveness, possibly mediated by interferon gamma. PMID- 9847281 TI - Ventilatory and cardiovascular responses to inspired He-O2 during exercise in chronic obstructive pulmonary disease. AB - Blunted maximum cardiac output and systemic O2 extraction could constitute primary limits to exercise in severe chronic obstructive pulmonary disease (COPD) or they could simply reflect cessation of exercise because of abnormal pulmonary mechanics. To determine which is the case, eight consecutive patients with severe COPD (FEV1 = 0. 56 +/- 0.04 L, mean +/- SEM), five of whom had alpha1 antiprotease deficiency, performed two incremental cycling tests while breathing N2-O2 or He-O2. Expired gases and V E were measured, and radial and pulmonary arterial blood was simultaneously sampled each minute. Peak exercise V E was higher with He-O2 than with N2-O2 (25.5 +/- 2. 2 versus 19.3 +/- 1.5 L/min, p = 0.002) and PaCO2 was lower (42 +/- 2 versus 46 +/- 2 mm Hg, p = 0.0003). V O2max improved only modestly (594 +/- 75 versus 514 +/- 54 ml/min, p = 0.04), and was accompanied by an increase in peak exercise CaO2 (18.7 +/- 0.9 versus 17.6 +/- 0. 9 ml/dl, p = 0.02). Peak Fick cardiac output was decreased (39 +/- 3% pred) and CvO2 was elevated (130 +/- 10% pred), and neither improved with He-O2 (p > 0.05 for each). Abnormal peak exercise cardiac output and systemic O2 extraction in severe COPD cannot be fully accounted for by limiting pulmonary mechanics and may contribute to exercise intolerance. PMID- 9847282 TI - Role of inducible nitric oxide synthase in endotoxin-induced acute lung injury. AB - The role of nitric oxide (NO) in lung injury remains unclear. Both beneficial and detrimental roles have been proposed. In this study, we used mutant mice lacking the inducible nitric oxide synthase (iNOS) to assess the role of this isoform in sepsis-associated lung injury. Wild-type and iNOS knockout mice were injected with either saline or Escherichia coli endotoxin (LPS) 25 mg/kg and killed 6, 12, and 24 h later. Lung injury was evaluated by measuring lactate dehydrogenase activity in the bronchoalveolar lavage, pulmonary wet/dry ratio, and immunostaining for nitrotyrosine formation. In the wild-type mice, LPS injection elicited more than a 3-fold rise in lactate dehydrogenase activity, a significant rise in lung wet/dry ratio and extensive nitrotyrosine staining in large airway and alveolar epithelium, macrophages, and pulmonary vascular cells. This was accompanied by induction of iNOS protein and increased lung nitric oxide synthase activity. By comparison, LPS injection in iNOS knockout mice elicited no iNOS induction and no significant changes in lung NOS activity, lactate dehydrogenase activity, lung wet/dry ratio, or pulmonary nitrotyrosine staining. These results indicate that mice deficient in iNOS gene are more resistant to LPS-induced acute lung injury than are wild-type mice. PMID- 9847284 TI - Differences in airway closure between normal and asthmatic subjects measured with single-photon emission computed tomography and technegas. AB - The absence of a maximal dose-response plateau as well as gas trapping and increases in closing capacity (CC) suggest that increased airway closure is an important mechanical abnormality of asthmatic airways. We compared the extent and distribution of airway closure in 13 normal and in 23 asthmatic subjects. Airway closure (LVclosed) was measured with single-photon emission computed tomography (SPECT) and an inhaled Technegas bolus as the percentage of lung volume without Technegas (LVtrans), and with CC, using nitrogen washout. LVclosed was compared in the apical, middle and lower zones, each being of equal vertical height. Values of mean LVclosed +/- 95% confidence interval (CI) were similar in normal (30 +/- 6.0% LVtrans) and asthmatic subjects (30 +/- 7.8% LVtrans). In normal subjects, LVclosed correlated with both age (r = 0.89, p < 0. 01) and CC (r = 0.86, p < 0.01), was more extensive in the lower zone (58 +/- 18.8% LVtrans, p < 0.01) than in the middle and upper zones (17 +/- 8.7% and 26 +/- 8.2 LVtrans, respectively), and increased with age in both the middle and lower zones (r = 0.94 and r = 0.90, respectively, p < 0.01). In asthmatic subjects, LVclosed did not correlate with age; was greatest in the lower zone, intermediate in the middle zone, and lowest in the apical zone (59 +/- 13.2%, 22 +/- 5.8%, and 12 +/- 4.4% LVtrans, respectively, p < 0. 01); and correlated weakly with age in the middle zone only (r = 0. 46, p < 0.05). We conclude that there is a predictable pattern of airway closure in normal subjects and that it is primarily influenced by pulmonary elastic recoil. This pattern is lost in asthmatic subjects. This may be explained by an increased range of closing pressures and a patchy distribution of airway closure, probably secondary to allergic inflammation. PMID- 9847283 TI - Artificial surfactant (Surfactant TA) modulates adherence and superoxide production of neutrophils. AB - Neutrophils cause lung injuries by releasing proteases and active oxygen radicals in patients with acute respiratory distress syndrome (ARDS). Artificial surfactant is used to replace native surfactant whose functions are deteriorated by serum-derived inhibitors in these patients. We investigated potential interactions between exogenous surfactant (Surfactant TA) and neutrophils in in vivo and in vitro experimental models. Neutrophil alveolitis was induced in hamster lungs by the intratracheal administration of bleomycin (5 mg/kg) on Day 0. Some of the animals were followed by replacement with Surfactant TA (5 and 10 mg/100 g body weight) on Day 1. Alveolar cells were harvested by lung lavage on Day 2. The numbers of the neutrophils obtained from the lungs treated with bleomycin and Surfactant TA were unchanged, but the superoxide production from these cells was significantly decreased when compared with control animals (no Surfactant TA). From the in vitro experiments, Surfactant TA was shown to inhibit adherence and superoxide production of human neutrophils. These effects were derived from the heat-resistant components of Surfactant TA and were mimicked by treatment with liposomes of dipalmitoyl phosphatidylcholine. Surfactant-TA treated neutrophils were demonstrated to have picnotic nuclei and to express Fas antigens, which were characteristic of apoptotic cells. These results suggest that exogenous Surfactant TA may play an important role not only in improving surfactant functions but in preventing neutrophils from further activation, probably through enhancing apoptosis. PMID- 9847285 TI - Mineral dusts directly induce epithelial and interstitial fibrogenic mediators and matrix components in the airway wall. AB - Exposure to mineral dusts is associated with the development of chronic airflow obstruction, probably mediated in part by dust-induced fibrosis of the small airways. To investigate the mechanism of fibrosis, we exposed rat tracheal explants to amosite asbestos, iron oxide, or titanium dioxide. Explants were then maintained in air organ culture, and the expression of genes encoding for various mediators and matrix components assessed by reverse transcriptase-polymerase chain reaction (RT-PCR). At 7 d, all dusts produced significant increases in platelet-derived growth factor-A (PDGF-A) and transforming growth factor-beta1 (TGF-beta1) gene expression compared with control; asbestos and titanium dioxide produced increases in PDGF-B, and titanium dioxide increased TGF-alpha expression. Only asbestos caused increases in procollagen expression. No dust increased expression of tumor necrosis factor-alpha (TNF-alpha), fibronectin, or tropoelastin. Elevations in these factors coincided temporally with transport of particles into the epithelium and then to the subepithelial space. By in situ hybridization, TGF-beta gene expression was found in both the epithelium and subepithelial (interstitial) space, and PDGF-B and procollagen gene expression in the subepithelial space. Chemical analysis showed a small increase in hydroxyproline, a measure of collagen content, in asbestos-treated explants. We conclude that mineral dusts can induce airway wall fibrosis by directly upregulating proliferative and fibrogenic mediators as well as matrix components in the airway epithelium and interstitium, and that neither airspace nor circulating inflammatory cells are required for these effects. Different mineral dusts produce different patterns of reaction. PMID- 9847286 TI - Investigation of effects of anesthesia and age on aspiration in mice through LacZ gene transfer by recombinant E1-deleted adenovirus vectors. AB - To examine the role of disturbed upper airway reflexes in aspiration, we administered 20 microliters of the adenovirus (Ad) vector Ad-CMV-LacZ or 20 microliters of phosphate buffered saline (PBS) intranasally to C57 black mice. We investigated expression of the LacZ gene by this Ad vector in the nostrils of each mouse, with or without anesthesia. Under anesthesia, LacZ gene expression was detected in the lungs of every mouse given the Ad vector. However, no LacZ gene expression was found in the trachea or lungs of mice given the Ad vector without anesthesia. In mice given PBS or wild-type adenovirus transnasally during anesthesia, there was no LacZ gene expression in the nostrils, trachea, or lungs, suggesting that with 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining, blue-stained cells indicated transferred LacZ gene expression. These results suggested that aspiration of intranasal solution into lower airways was caused by disturbed upper airway reflexes during anesthesia. This process can be analyzed by the distribution of LacZ gene expression in airways. We next examined the effect of age on anesthesia-induced aspiration. Twenty-six-mo-old mice exhibited more LacZ gene expression in their lungs than did 6-mo-old mice at a concentration of 0.5 to 4.0% halothane in 100% oxygen. This suggests that light anesthesia may depress upper airway reflexes and cause aspiration in older animals. This novel model of aspiration, generated with the Ad-CMV-LacZ vector, may be useful for elucidating the mechanism of development of aspiration pneumonia in relation to age-related impairment of upper airway reflexes. PMID- 9847287 TI - Alveolar hypoxia increases gene expression of extracellular matrix proteins and platelet-derived growth factor-B in lung parenchyma. AB - The walls of pulmonary capillaries are extremely thin, and wall stress increases greatly when capillary pressure rises. Alveolar hypoxia causes pulmonary vasoconstriction and hypertension, and if this is uneven, some capillaries may be exposed to high transmural pressure and develop stress failure. There is evidence that increased wall stress causes capillary remodeling. In this study we exposed Madison strain Sprague-Dawley rats to normobaric hypoxia (10% oxygen) for 6 h or 3 d (short-term group), and for 3 d or 10 d (long-term group). Peripheral lung tissue was then collected and messenger RNA (mRNA) levels were determined for extracellular matrix (ECM) proteins and growth factors. Collagen content (hydroxyproline) was also measured. Levels of mRNA for alpha2(IV) procollagen increased sixfold after 6 h of hypoxia and sevenfold after 3 d of hypoxia, and then decreased after 10 d exposure. Levels of mRNA for platelet-derived growth factor-B (PDGF-B) doubled after 6 h of hypoxia but returned to control values after 3 d. mRNA levels for alpha1(I) and alpha1(III) procollagens and fibronectin were increased after 3 d of hypoxia (by seven- to 12-fold, 1.6- to eightfold, and 12-fold, respectively), then decreased toward control values after 10 d. In contrast, neither levels of mRNA for vascular endothelial growth factor (VEGF) nor collagen content changed. These results suggest that alveolar hypoxia causes vascular remodeling in lung parenchyma, and are consistent with capillary wall remodeling in response to increased wall stress. PMID- 9847288 TI - Bronchiolitis obliterans organizing pneumonia syndrome primed by radiation therapy to the breast. The Groupe d'Etudes et de Recherche sur les Maladies Orphelines Pulmonaires (GERM"O"P) AB - Reports of bronchiolitis obliterans organizing pneumonia (BOOP) occurring in women after radiation therapy for breast cancer have suggested that radiation to the lung could participate in the development of BOOP. We now describe the clinical, radiographic, functional, and bronchoalveolar lavage characteristics of this syndrome in a series of 15 patients reported to the Groupe d'Etudes et de Recherche sur les Maladies "Orphelines" Pulmonaires (GERM"O"P) in France. All 15 women (60 +/- 6 yr of age) fulfilled the following inclusion criteria: (1) radiation therapy to the breast within 12 mo, (2) general and/or respiratory symptoms lasting for at least 2 wk, (3) lung infiltrates outside the radiation port, and (4) no specific cause. The patients presented with fever, nonproductive cough, mild dyspnea, and peripheral alveolar opacities on chest radiograph with a characteristic migratory pattern. In five patients, BOOP was found at lung pathologic analysis. In all the patients dramatic improvement was obtained with corticosteroids, but relapses occurred in 12 patients while tapering or after stopping corticosteroids. This report demonstrates that a characteristic BOOP syndrome may occur after radiation therapy to the breast, including tangential radiation to the lung, thus suggesting that radiation therapy may prime the development of BOOP. PMID- 9847289 TI - Collagen deposition in large airways may not differentiate severe asthma from milder forms of the disease. AB - Chronic airway inflammation and remodeling, including fibrosis, have been proposed as important contributors to asthma pathophysiology. Previous studies of airway fibrosis have been performed mainly in mild and moderate asthmatics at the subepithelial "basement membrane" (SBM) level. The current study was designed to evaluate the large airway SBM thickness and submucosal collagen deposition, as measured by three different collagen staining methods, in endobronchial biopsies from 17 severe, nine moderate, and seven mild asthmatics, as well as eight normal control subjects. Tissue eosinophils and transforming growth factor-beta (TGF beta) immunoreactivity were also examined. There were no statistically significant differences in the SBM thickness, submucosal collagen deposition, eosinophil numbers, or TGF-beta positive cells among the three groups of asthmatics and the normal control subjects. It was only when examining all asthmatics (n = 33) together, that a modestly thickened SBM (p = 0.04), as evaluated by collagen type III immunostaining, was observed as compared with normal control subjects. Despite this difference, no significant differences were found in the amount of submucosal collagen deposition and the number of eosinophils or TGF-beta expressing cells when comparing total asthmatics and normal control subjects. Additionally, no significant correlations were found between collagen deposition and eosinophil count, TGF-beta expression level, FEV1, or duration of asthma. These results suggest that although increased collagen deposition in the SBM at the large airway level is a characteristic of asthma, it may not explain the differences in severity of asthma. PMID- 9847290 TI - Sputum metalloproteinase-9/tissue inhibitor of metalloproteinase-1 ratio correlates with airflow obstruction in asthma and chronic bronchitis. AB - Asthma and chronic bronchitis are inflammatory diseases with extracellular matrix (ECM) remodeling and collagen deposition. Collagen homeostasis is controlled by metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). We evaluated MMP and TIMP balance in induced sputum of 10 control, 31 untreated asthmatic, and 16 chronic bronchitic subjects. We first performed zymographic analysis to identify the profile of MMPs. Zymography revealed a similar MMPs profile in all populations studied and that MMP-9 was the major enzyme released. We then measured, using enzyme immunoassay, the concentrations of MMP-9 and of its inhibitor TIMP-1 and evaluated whether airflow limitation may be associated with an imbalance between these enzymes. MMP-9 and TIMP-1 concentrations were greater in sputum of patients with asthma and chronic bronchitis than in control subjects. The molar ratio between MMP-9 and TIMP-1 was lower in asthmatics and chronic bronchitics than in control subjects, and positively correlated with FEV1 values. In asthma, MMP-9 levels were significantly correlated with the number of macrophages and neutrophils. This study shows that airway inflammation in asthma and chronic bronchitis is associated with an imbalance between MMP-9 and TIMP-1 which may have a role in the pathogenesis of ECM remodeling and airflow obstruction. PMID- 9847291 TI - Transforming growth factor beta1 and recruitment of macrophages and mast cells in airways in chronic obstructive pulmonary disease. AB - Chronic airways inflammation is one of the features of chronic obstructive pulmonary disease (COPD). We demonstrated previously that bronchiolar epithelium in COPD contains increased numbers of macrophages and mast cells. Transforming growth factor beta1 (TGF-beta1) may be involved in this influx because it has chemotactic activity for macrophages and mast cells. In this study, we examined expression patterns of TGF-beta1, TGF-beta receptors type I and II (TGF-betaRI and TGF-betaRII) by immunohistochemistry and mRNA in situ hybridization in peripheral lung tissue of 14 current or ex-smokers with COPD (FEV1 < 75%) and 14 without COPD (FEV1 > 84%). In both groups, TGF-beta1 and its receptors are present in airway and alveolar epithelial cells, airway and vascular smooth muscle cells, and tissue and alveolar CD68(+) cells (considered herein to be macrophages). In subjects with COPD, a semiquantitative analysis revealed approximately twofold higher levels of TGF-beta1 mRNA and protein in bronchiolar and alveolar epithelium (p < 0.02) as compared with subjects without COPD. With regard to bronchiolar epithelial cells, we found a significant correlation between TGF-beta1 mRNA and protein expression (r = 0.62; p < 0.002), and between the FEV1 of all subjects together and TGF-beta1 protein (r = -0.60; p < 0.0002) and mRNA (r = -0.67; p < 0. 002) levels. The epithelial expression of TGF-beta1 mRNA and TGF-beta1 protein correlates with the number of intraepithelial macrophages (both: r = 0.44; p < 0.03) whereas intraepithelial mast cell numbers correlate with epithelial TGF-beta1 mRNA expression. These data suggest a role for TGF-beta1 in recruiting macrophages into the airway epithelium in COPD. PMID- 9847292 TI - Interleukin-10 and transforming growth factor-beta promoter polymorphisms in allergies and asthma. AB - Interleukin-10 (IL-10) and transforming growth factor beta (TGF-beta) are inhibitory for B and T cells, IgE production, and mast cell proliferation, and they induce apoptosis in eosinophils. These cytokines are therefore candidate genes which could contribute to the development of asthma or allergies. We investigated the hypothesis that polymorphic nucleotides within the IL-10 and TGF beta gene promoters would link to the expression of allergies and asthma. DNA taken from families with an asthmatic proband was examined for base exchanges by single-stranded conformational polymorphism (SSCP). We demonstrated the presence of a polymorphism in the promoter region of the IL-10 gene and four in the TGF beta gene promoters (3 in TGF-beta1 and 1 in TGF-beta2). The IL-10 gene polymorphism was a C-to-A exchange 571 base pairs upstream from the translation start site and was present between consensus binding sequences for Sp1 and elevated total serum. This polymorphism was associated with elevated total serum IgE in subjects heterozygotic or homozygotic for this base exchange (p < 0.009). The base exchange at -509 (from the transcription initiation site) in the TGF beta promoter also linked to elevated total IgE (p < 0.01). This polymorphism represented a C-to-T base exchange which induced a YY1 consensus sequence and is present in a region of the promoter associated with negative transcription regulation. PMID- 9847294 TI - Association of persistent bronchial hyperresponsiveness with beta2-adrenoceptor (ADRB2) haplotypes. A population study. AB - Bronchial hyperresponsiveness (BHR) is a hallmark of asthma and represents a strong risk factor for the disease. However, not all asthmatics have BHR and it can be observed in normal subjects too, probably because of genetic predisposition. Increasing attention is being focused on the beta2-adrenoceptor gene (ADRB2), whose genetic variability at amino acids 16 and 27 has been shown to correlate with some clinical features of asthma, including airways reactivity. To verify whether ADRB2 gene polymorphisms can influence BHR at a broader level, we studied a large, highly homogeneous sample of individuals sharing race, gender, age, and current living environment. BHR was strictly defined as a constant positive response to serial methacholine challenge tests and an improved definition of genetic variability at the ADRB2 locus was used, by identifying the haplotypic combinations of polymorphisms 16 and 27. We observed that the ADRB2 haplotype with a Gly at position 16 and a Gln at position 27 is associated with BHR in our sample. The association persisted also after correction for potentially confounding variables such as specific and total IgE levels. This observation suggests therefore that ADRB2 gene can confer genetic susceptibility to BHR, rather than having only a disease-modifying effect in asthma. PMID- 9847293 TI - Systolic ventricular dysfunction causes selective diaphragm atrophy in rats. AB - In order to examine the relative impairment of the diaphragm and other skeletal muscles in systolic ventricular dysfunction (VD), their structure and function were compared between rats with VD induced by left coronary artery ligation (n = 17) and sham-operated rats (Co, n = 10). In addition, in an attempt to unravel the mechanism of the observed impairment, we examined alterations in insulin-like growth factor-I (IGF-I) serum levels and IGF-I expression in the liver, diaphragm, and gastrocnemius. In a second series of rats (VD, n = 5 and Co, n = 5) hemodynamic measurements were performed. All measurements were performed 3 mo after the operation. Infarct size averaged 32 +/- 10 and 44 +/- 20% in the two series, respectively (NS). Hemodynamic measurements revealed a decrease in left ventricular peak systolic pressure of 19% (p < 0. 05). Significant diaphragm atrophy (weight: 622 +/- 52 mg in VD versus 750 +/- 54 mg in Co, p < 0.0005), without alterations in diaphragm contractile properties was present in VD animals. For all animals combined, the reduction in diaphragm weight was related to infarct size (r = -0.74, p < 0.001). No alterations were observed in the other inspiratory and peripheral muscles. ATPase staining of the diaphragm showed atrophy of type I and type IIx/b fibers, their cross-sectional area (CSA) being reduced by 13 and 16%, respectively (p < 0.05). There were no signs of myopathic alterations. IGF-I expression was increased by 55% in the diaphragm of rats with VD (p < 0.05). IGF-I expression in the liver and gastrocnemius and serum IGF-I levels were unaltered. These data suggest the presence of compensatory mechanisms aimed at minimizing diaphragmatic fiber atrophy. We conclude that systolic VD caused: (1) selective diaphragm atrophy, which was related to infarct size; (2) a decrease in diaphragm type I and IIx/b CSA not associated with myopathic changes; (3) an increase in the IGF-I mRNA content of the diaphragm. The selective diaphragm involvement in the present study may be related to the moderate degree of ventricular dysfunction induced. PMID- 9847295 TI - Progressive retropalatal narrowing preceding obstructive apnea. AB - Pharyngeal occlusion during obstructive apnea is thought to be an inspiratory related event; however, occlusion also occurs in the absence of negative intrathoracic pressure. We hypothesized that inspiratory-related pharyngeal occlusion would be preceded by significant expiratory narrowing. Eight sleeping patients with obstructive apnea were studied. Pharyngeal caliber, airflow, and esophageal pressure (Pes) were simultaneously monitored during three to four consecutive breaths preceding occlusion (between 3 and 22 events were studied per subject). Relative changes in retropalatal airway cross-sectional area (CSA) were determined from fiberoptic images (five frames per second) normalized to the maximum CSA. During inspiration, CSA was significantly reduced only during the breath immediately preceding the apnea (Group mean CSA +/- SEM: 51 +/- 8% at the start of inspiration compared with 37 +/- 8% at midinspiration). During expiration, for all breaths there was an initial significant increase in CSA compared with the nadir CSA during the preceding inspiration (CSA: breath-3, 57 +/- 10% to 79 +/- 3%; breath-2, 59 +/- 8% to 76 +/- 4%; breath-1, 37 +/- 8% to 64 +/- 8%), followed by a significant narrowing at end-expiration compared with the peak CSA during that expiration (CSA: breath-3, 79 +/- 3% to 62 +/- 6%; breath-2, 76 +/- 4% to 50 +/- 10%; breath-1, 64 +/- 8% to 36 +/- 10%). Occlusion occurred at a pressure significantly less than that generated during the previous unoccluded breath (Pes: breath-1, -10.8 +/- 2.9 cm H2O; occlusion, -8.2 +/- 1.9 cm H2O). These results show that expiratory narrowing produced a significant reduction of CSA at end-expiration prior to obstructive apnea. PMID- 9847296 TI - Abnormal surfactant metabolism and function in preterm ventilated baboons. AB - We evaluated surfactant metabolism and function and the effects of antenatal glucocorticoids in very preterm baboons. Pregnant baboons were randomized to receive saline (controls) or 6 mg betamethasone (beta) 48 and 24 h before delivery at 125 +/- 2 d gestation (term is 184 d). The newborn baboons were treated with [14C]dipalmitoylphosphatidylcholine-labeled surfactant and ventilated for 6 d. Lung function for six control and six betamethasone-treated animals was similar. Recoveries of 14C-saturated phosphatidylcholine (Sat PC) were similar: 4.8% (control) and 3.6% (beta) in alveolar wash and 15.4% (control) and 17.7% (beta) in total lungs. Alveolar and total lung pool sizes of Sat PC were about 23 and 190 micromol/kg, respectively. The preterm baboons secreted 8.7% (control) and 6.5% (beta) of de novo synthesized Sat PC labeled with 3H palmitate from Day 5 to Day 6. These preterm baboons had high estimated Sat PC synthetic and net tissue accumulation rates but low secretion of Sat PC. The large aggregate surfactant fractions from the preterm baboons had high minimal surface tensions and were less effective when used to treat surfactant-deficient preterm rabbits than surfactant from newborn or adult baboons. Ventilation of the preterm baboon was associated with surfactant functional and metabolic abnormalities that were not altered by antenatal glucocorticoids. PMID- 9847297 TI - Reversal of hypocalcemia and decreased afterload in sepsis. Effect on myocardial systolic and diastolic function. AB - Sepsis is a major cause of death in intensive care units. Clinically, sepsis induces a number of physiologic and metabolic abnormalities, including decreased myocardial contractility and decreased plasma ionized calcium. There is debate about the proper therapy of hypocalcemia in sepsis because calcium administration may worsen cell function by causing intracellular Ca2+ overload. We investigated the effect of Ca2+ administration on myocardial systolic and diastolic function in an extensively utilized rat model of sepsis, i.e., the cecal ligation and puncture model (CLP). Approximately 24 h after CLP or sham surgery, rats were anesthetized and myocardial function assessed in vivo by a left ventricular Millar catheter and simultaneous two-dimensional guided M-mode echocardiography. Septic rats had a 28% decrease in peak left ventricular developed pressure, a 30% decrease in +dP/ dt, and a 23% decrease in -dP/dt (p < 0.05). Plasma ionized Ca2+ was decreased in septic compared with that in sham rats: 4.9 +/- 0.9 and 5.6 +/- 0.01 mg/dl, respectively (p < 0.05). CaCl2 improved both systolic and diastolic function and there was no evidence of adverse effects of Ca2+ even at supraphysiologic levels. Surprisingly, correction of decreased afterload in septic rats, using the pure alpha-agonist phenylephrine, caused normalization of all indices of cardiac contractility, indicating that the presumed decrease in cardiac function was due entirely to an effect of the decreased afterload to "unload" the left ventricle. We conclude that Ca2+ administration is not detrimental to cardiac function in the rat CLP model. Although the rat CLP model is widely utilized and reproduces many of the clinical hallmarks of sepsis, it does not cause intrinsic myocardial depression and, therefore, it may not be an appropriate model to investigate the clinical cardiac dysfunction that occurs in patients with sepsis. PMID- 9847298 TI - Monoclonal endothelial cells in appetite suppressant-associated pulmonary hypertension. AB - Anorexigens such as aminorex fumarate and dexfenfluramine are associated with the development of severe pulmonary hypertension (PH), which clinically and histopathologically is considered indistinguishable from idiopathic or primary pulmonary hypertension (PPH). For the current study, we asked whether anorexigen associated PH is characterized by monoclonal pulmonary endothelial cell proliferation (such as in PPH) or, alternatively, is associated with a polyclonal endothelial cell proliferation as found in secondary PH. Analysis of clonality by the human androgen receptor assay was performed in microdissected endothelial cells of plexiform lesions of two patients with anorexigen-associated PH. The four plexiform lesions of Patient 1 and the six of Patient 2 with anorexigen associated PH exhibited a monoclonal expansion of pulmonary endothelial cells, with a mean clonality ratio of 0.03 +/- 0.01 SE. Our results indicate that appetite suppressant-associated PH is identical to PPH not only in clinical and histopathologic features but also, at a molecular level, in terms of the monoclonal nature of the endothelial cell proliferation. The anorexigens may accelerate the growth of pulmonary endothelial cells in patients with predisposition to develop PPH. PMID- 9847299 TI - Neurotrophins are increased in bronchoalveolar lavage fluid after segmental allergen provocation. AB - The mechanisms linking inflammation and airway hyperresponsiveness in allergic bronchial asthma are still not completely defined. Since neurotrophic factors increase nerve excitability and neurotransmitter synthesis and are produced by immunocompetent cells, they are likely candidates as mediators of inflammation and hyperresponsiveness. We tested the hypothesis that neurotrophin concentrations will increase in the bronchoalveolar lavage (BAL) fluid from patients with asthma after segmental allergen provocation. For this purpose an individually standardized dose of allergen or saline was instilled into different segments during bronchoscopy in eight subjects with mild allergic bronchial asthma. Segments were then lavaged 10 min and 18 h after allergen challenge or saline instillation. There was a significant increase in the neurotrophins nerve growth factor, brain-derived neurotrophic factor, and neurotrophin-3 in BAL fluids 18 h after allergen but not saline challenge. We conclude that neurotrophins are produced endobronchially following allergen provocation, suggesting a contribution to the pathogenesis of asthma. PMID- 9847300 TI - AIDS vaccination studies using an ex vivo feline immunodeficiency virus model: detailed analysis of the humoral immune response to a protective vaccine. AB - The feline immunodeficiency virus (FIV) cat model is extensively used to investigate possible vaccination approaches against AIDS in humans. Although consistent levels of protection have been achieved with FIV, as with other model systems, by immunizing with whole inactivated virus or fixed infected cells, the mechanisms responsible for protection are elusive. In previous studies we showed that cats immunized with a vaccine consisting of fixed infected cells were protected or unprotected against cell-free or cell-associated FIV challenge depending on the time interval between completion of vaccination and challenge. In an attempt to define possible humoral immune correlates of protection, selected sera harvested at the times of challenge from such cats were examined for anti-FIV-antibody titers and properties by using binding and functional immunological assays. Binding assays included quantitative Western blotting, enzyme-linked tests for antibodies to FIV glycoproteins and immunodominant linear epitopes, and tests for measuring conformation dependence and avidity of anti viral-envelope antibodies. Functional assays included virus neutralization performed with two different cell substrates, complement- and antibody-dependent virolysis, blocking of reverse transcriptase, and an assay that measured the ability of sera to prevent FIV growth in cocultures of infected and uninfected cells. Despite the wide spectrum of parameters investigated, no correlation between vaccine-induced protection and the humoral parameters measured was noted. PMID- 9847301 TI - Identification of a domain containing B-cell epitopes in hepatitis C virus E2 glycoprotein by using mouse monoclonal antibodies. AB - Evidence from clinical and experimental studies of human and chimpanzees suggests that hepatitis C virus (HCV) envelope glycoprotein E2 is a key antigen for developing a vaccine against HCV infection. To identify B-cell epitopes in HCV E2, six murine monoclonal antibodies (MAbs), CET-1 to -6, specific for HCV E2 protein were generated by using recombinant proteins containing E2t (a C terminally truncated domain of HCV E2 [amino acids 386 to 693] fused to human growth hormone and glycoprotein D). We tested whether HCV-infected sera were able to inhibit the binding of CET MAbs to the former fusion protein. Inhibitory activity was observed in most sera tested, which indicated that CET-1 to -6 were similar to anti-E2 antibodies in human sera with respect to the epitope specificity. The spacial relationship of epitopes on E2 recognized by CET MAbs was determined by surface plasmon resonance analysis and competitive enzyme linked immunosorbent assay. The data indicated that three overlapping epitopes were recognized by CET-1 to -6. For mapping the epitopes recognized by CET MAbs, we analyzed the reactivities of CET MAbs to six truncated forms and two chimeric forms of recombinant E2 proteins. The data suggest that the epitopes recognized by CET-1 to -6 are located in a small domain of E2 spanning amino acid residues 528 to 546. PMID- 9847303 TI - Function of a bovine papillomavirus type 1 exonic splicing suppressor requires a suboptimal upstream 3' splice site. AB - Alternative splicing is an important mechanism for the regulation of bovine papillomavirus type 1 (BPV-1) gene expression during the virus life cycle. Previous studies in our laboratory have identified two purine-rich exonic splicing enhancers (ESEs), SE1 and SE2, located between two alternative 3' splice sites at nucleotide (nt) 3225 and nt 3605. Further analysis of BPV-1 late-pre mRNA splicing in vitro revealed a 48-nt pyrimidine-rich region immediately downstream of SE1 that inhibits utilization of the nt 3225 3' splice site. This inhibitory element, which we named an exonic splicing suppressor (ESS), has a U rich 5' end, a C-rich central part, and an AG-rich 3' end (Z. M. Zheng, P. He, and C. C. Baker, J. Virol. 70:4691-4699, 1996). The present study utilized in vitro splicing of both homologous and heterologous pre-mRNAs to further characterize the ESS. The BPV-1 ESS was inserted downstream of the 3' splice site in the BPV-1 late pre-mRNA, Rous sarcoma virus src pre-mRNA, human immunodeficiency virus tat-rev pre-mRNA, and Drosophila dsx pre-mRNA, all containing a suboptimal 3' splice site, and in the human beta-globin pre-mRNA, which contains a constitutive 3' splice site. These studies demonstrated that suppression of splicing by the BPV-1 ESS requires an upstream suboptimal 3' splice site but not an upstream ESE. Furthermore, the ESS functions when located either upstream or downstream of BPV-1 SE1. Mutational analyses demonstrated that the function of the ESS is sequence dependent and that only the C-rich region of the ESS is essential for suppression of splicing in all the pre-mRNAs tested. PMID- 9847302 TI - Mutational analysis of the hydrophobic tail of the human immunodeficiency virus type 1 p6(Gag) protein produces a mutant that fails to package its envelope protein. AB - The p6(Gag) protein of human immunodeficiency virus type 1 (HIV-1) is produced as the carboxyl-terminal sequence within the Gag polyprotein. The amino acid composition of this protein is high in hydrophilic and polar residues except for a patch of relatively hydrophobic amino acids found in the carboxyl-terminal 16 amino acids. Internal cleavage of p6(Gag) between Y36 and P37, apparently by the HIV-1 protease, removes this hydrophobic tail region from approximately 30% of the mature p6(Gag) proteins in HIV-1MN. To investigate the importance of this cleavage and the hydrophobic nature of this portion of p6(Gag), site-directed mutations were made at the minor protease cleavage site and within the hydrophobic tail. The results showed that all of the single-amino-acid replacement mutants exhibited either reduced or undetectable cleavage at the site yet almost all were nearly as infectious as wild-type virus, demonstrating that processing at this site is not important for viral replication. However, one exception, Y36F, was 300-fold as infectious the wild type. In contrast to the single-substitution mutants, a virus with two substitutions in this region of p6(Gag), Y36S-L41P, could not infect susceptible cells. Protein analysis showed that while the processing of the Gag precursor was normal, the double mutant did not incorporate Env into virus particles. This mutant could be complemented with surface glycoproteins from vesicular stomatitis virus and murine leukemia virus, showing that the inability to incorporate Env was the lethal defect for the Y36S L41P virus. However, this mutant was not rescued by an HIV-1 Env with a truncated gp41(TM) cytoplasmic domain, showing that it is phenotypically different from the previously described MA mutants that do not incorporate their full-length Env proteins. Cotransfection experiments with Y36S-L41P and wild-type proviral DNAs revealed that the mutant Gag dominantly blocked the incorporation of Env by wild type Gag. These results show that the Y36S-L41P p6(Gag) mutation dramatically blocks the incorporation of HIV-1 Env, presumably acting late in assembly and early during budding. PMID- 9847304 TI - Targeting of the visna virus tat protein to AP-1 sites: interactions with the bZIP domains of fos and jun in vitro and in vivo. AB - The visna virus Tat protein is required for efficient viral transcription from the visna virus long terminal repeat (LTR). AP-1 sites within the visna virus LTR, which can be bound by the cellular transcription factors Fos and Jun, are also necessary for Tat-mediated transcriptional activation. A potential mechanism by which the visna virus Tat protein could target the viral promoter is by protein-protein interactions with Fos and/or Jun bound to AP-1 sites in the visna virus LTR. Once targeted to the visna virus promoter, the Tat protein could then interact with basal transcription factors to activate transcription. To examine protein-protein interactions with cellular proteins at the visna virus promoter, we used an in vitro protein affinity chromatography assay and electrophoretic mobility shift assay, in addition to an in vivo two-hybrid assay, to show that the visna virus Tat protein specifically interacts with the cellular transcription factors Fos and Jun and the basal transcription factor TBP (TATA binding protein). The Tat domain responsible for interactions with Fos and Jun was localized to an alpha-helical domain within amino acids 34 to 69 of the protein. The TBP binding domain was localized to amino acids 1 to 38 of Tat, a region previously described by our laboratory as the visna virus Tat activation domain. The bZIP domains of Fos and Jun were found to be important for the interactions with Tat. Mutations within the basic domains of Fos and Jun abrogated binding to Tat in the in vitro assays. The visna virus Tat protein was also able to interact with covalently cross-linked Fos and Jun dimers. Thus, the visna virus Tat protein appears to target AP-1 sites in the viral promoter in a mechanism similar to the interaction of human T-cell leukemia virus type 1 Tax with the cellular transcription factor CREB, by binding the basic domains of an intact bZIP dimer. The association between Tat, Fos, and Jun would position Tat proximal to the viral TATA box, where the visna virus Tat activation domain could contact TBP to activate viral transcription. PMID- 9847305 TI - Influenza C virus CM2 protein is produced from a 374-amino-acid protein (P42) by signal peptidase cleavage. AB - Although unspliced mRNA from influenza C virus RNA segment 6 (M gene) has a single open reading frame capable of encoding a 374-amino-acid protein (Mr, 42,000), the major polypeptide synthesized from this mRNA species is the CM2 protein, with an Mr of 18,000. The present study was performed to investigate the mechanism by which CM2 is generated from the unspliced mRNA. It was reported previously that the 374-amino-acid protein (P42) is an integral membrane protein having two internal hydrophobic domains, one of which (residues 241 to 252) is followed by two sequences (252 Ile-Thr-Ser and 257 Ala-Ser-Ala) favorable for cleavage by signal peptidase. To examine the possibility that P42 is cleaved by signal peptidase after Ser residue 254 or Ala residue 259 to yield CM2, we constructed three mutated M gene cDNAs in which either or both of the two sequences were eliminated and tested their ability to synthesize CM2 in the transfected COS cells. The results showed that CM2 synthesis was blocked completely when the second recognition motif for signal peptidase was removed. It was also found that when the mRNA transcript of the wild-type M gene was translated in vitro, P42, but not CM2, was synthesized in the absence of dog pancreas microsomal membranes, whereas CM2, in addition to a polypeptide (designated M1') slightly larger than matrix protein (M1), was synthesized in the presence of microsomes. When the same experiment was done with the transcript of the mutated M gene in which the second recognition motif was removed, synthesis of CM2 could not be seen, even in the presence of microsomes. From these results, we conclude that cleavage of P42 by signal peptidase after Ala residue 259 produces CM2, composed of the C-terminal 115 amino acids, in addition to M1', composed of the N-terminal 259 amino acids. PMID- 9847306 TI - Spontaneous mutation rate of measles virus: direct estimation based on mutations conferring monoclonal antibody resistance. AB - High mutation rates typical of RNA viruses often generate a unique viral population structure consisting of a large number of genetic microvariants. In the case of viral pathogens, this can result in rapid evolution of antiviral resistance or vaccine-escape mutants. We determined a direct estimate of the mutation rate of measles virus, the next likely target for global elimination following poliovirus. In a laboratory tissue culture system, we used the fluctuation test method of estimating mutation rate, which involves screening a large number of independent populations initiated by a small number of viruses each for the presence or absence of a particular single point mutation. The mutation we focused on, which can be screened for phenotypically, confers resistance to a monoclonal antibody (MAb 80-III-B2). The entire H gene of a subset of mutants was sequenced to verify that the resistance phenotype was associated with single point mutations. The epitope conferring MAb resistance was further characterized by Western blot analysis. Based on this approach, measles virus was estimated to have a mutation rate of 9 x 10(-5) per base per replication and a genomic mutation rate of 1.43 per replication. The mutation rates we estimated for measles virus are comparable to recent in vitro estimates for both poliovirus and vesicular stomatitis virus. In the field, however, measles virus shows marked genetic stability. We briefly discuss the evolutionary implications of these results. PMID- 9847307 TI - Herpes simplex virus processivity factor UL42 imparts increased DNA-binding specificity to the viral DNA polymerase and decreased dissociation from primer template without reducing the elongation rate. AB - Herpes simplex virus DNA polymerase consists of a catalytic subunit, Pol, and a processivity subunit, UL42, that, unlike other established processivity factors, binds DNA directly. We used gel retardation and filter-binding assays to investigate how UL42 affects the polymerase-DNA interaction. The Pol/UL42 heterodimer bound more tightly to DNA in a primer-template configuration than to single-stranded DNA (ssDNA), while Pol alone bound more tightly to ssDNA than to DNA in a primer-template configuration. The affinity of Pol/UL42 for ssDNA was reduced severalfold relative to that of Pol, while the affinity of Pol/UL42 for primer-template DNA was increased approximately 15-fold relative to that of Pol. The affinity of Pol/UL42 for circular double-stranded DNA (dsDNA) was reduced drastically relative to that of UL42, but the affinity of Pol/UL42 for short primer-templates was increased modestly relative to that of UL42. Pol/UL42 associated with primer-template DNA approximately 2-fold faster than did Pol and dissociated approximately 10-fold more slowly, resulting in a half-life of 2 h and a subnanomolar Kd. Despite such stable binding, rapid-quench analysis revealed that the rates of elongation of Pol/UL42 and Pol were essentially the same, approximately 15 [corrected] nucleotides/s. Taken together, these studies indicate that (i) Pol/UL42 is more likely than its subunits to associate with DNA in a primer-template configuration rather than nonspecifically to either ssDNA or dsDNA, and (ii) UL42 reduces the rate of dissociation from primer-template DNA but not the rate of elongation. Two models of polymerase-DNA interactions during replication that may explain these findings are presented. PMID- 9847308 TI - Antigen-specific expansion of cytotoxic T lymphocytes in acute measles virus infection. AB - Skewing of the T-cell receptor repertoire of CD8(+) T cells has been shown in some persistent infections with viruses, such as human immunodeficiency virus, simian immunodeficiency virus, and Epstein-Barr virus. We have demonstrated that similar distortions also occur in nonpersistent measles virus infection. In addition, two of four children immunized with live, attenuated measles virus showed larger and more persistent CD8(+) T-cell expansions than their naturally infected counterparts. The expanded lymphocyte populations were monoclonal or oligoclonal and lysed target cells infected with recombinant vaccinia virus expressing measles virus protein. These results demonstrate that the expansions of CD8(+) T lymphocytes are antigen driven. PMID- 9847309 TI - A novel heterogeneous nuclear ribonucleoprotein-like protein interacts with NS1 of the minute virus of mice. AB - NS1, the major nonstructural parvovirus protein of the minute virus of mice, is a multifunctional protein responsible for several aspects of viral replication. NS1 transactivates the P38 promoter (used to express the structural proteins), as well as its own strong promoter, P4. To study the mechanism of activation and to map regions of NS1 responsible for transactivation, NS1 and various deletions of NS1 were cloned in frame with the GAL4DB and cotransfected into COS-7 and LA9 cells with a synthetic GAL4-responsive reporter plasmid. These studies showed NS1 can directly activate transcription through its 129 carboxyl-terminal amino acid residues. Any deletion from this region of the C terminus, even as few as 8 amino acids, completely abolishes transactivation. A yeast two-hybrid system used to identify protein-protein interactions demonstrated that NS1 is able to dimerize when expressed in yeast cells. However, only an almost complete NS11-638 bait was able to interact with the full-length NS1. A two-hybrid screen identified a HeLa cell cDNA clone (NS1-associated protein 1 [NSAP1]) that interacts with NS11-276 and NS11-638. An additional sequence was predicted from human EST (expressed sequence tag) data, and the cDNA was estimated to be at least 2,221 bp long, potentially encoding a 562-amino-acid protein product. A polyclonal antibody raised to a synthetic peptide within NSAP1 recognizes an approximately 65-kDa cellular protein. This NSAP1 cDNA has not previously been characterized, but the predicted protein sequence is 80% identical to the recently identified heterogeneous nuclear ribonucleoprotein (hnRNP) R (W. Hassfeld et al., Nucleic Acids Res. 26:439-445, 1998). NSAP1 contains four ribonucleoprotein domains, as well as a highly repetitive C-terminal region. A closely related mouse cDNA (deduced from murine EST data) encodes a protein with only a single amino acid residue change from the human protein. NSAP1 is predicted to be a 65-kDa polynucleotide binding protein, and it likely functions in the regulation of splicing and/or transport of mRNAs from the nucleus. PMID- 9847310 TI - A hairpin structure in the R region of the human immunodeficiency virus type 1 RNA genome is instrumental in polyadenylation site selection. AB - Some retroviruses with an extended repeat (R) region encode the polyadenylation signal within the R region such that this signal is present at both the 5' and 3' ends of the viral transcript. This necessitates differential regulation to either repress recognition of the 5' polyadenylation signal or enhance usage of the 3' signal. The human immunodeficiency virus type 1 (HIV-1) genome encodes an inherently efficient polyadenylation signal within the 97-nucleotide R region. Polyadenylation at the 5' HIV-1 polyadenylation site is inhibited by downstream splicing signals, and usage of the 3' polyadenylation site is triggered by an upstream enhancer element. In this paper, we demonstrate that this on-off switch of the HIV-1 polyadenylation signal is controlled by a secondary RNA structure that occludes part of the AAUAAA hexamer motif, which we have termed the polyA hairpin. Opening the 5' hairpin by mutation triggered premature polyadenylation and caused reduced synthesis of viral RNA, indicating that the RNA structure plays a pivotal role in repression of the 5' polyadenylation site. Apparently, the same hairpin structure does not interfere with efficient usage of the 3' polyadenylation site, which may be due to the presence of the upstream enhancer element. However, when the 3' hairpin was further stabilized by mutation, we measured a complete loss of 3' polyadenylation. Thus, the thermodynamic stability of the polyA hairpin is delicately balanced to allow nearly complete repression of the 5' site yet efficient activation of the 3' site. This is the first report of regulated polyadenylation that is mediated by RNA secondary structure. A similar hairpin motif that occludes the polyadenylation signal can be proposed for other lentiviruses and members of the spumaretroviruses, suggesting that this represents a more general gene expression strategy of complex retroviruses. PMID- 9847311 TI - Vpu increases susceptibility of human immunodeficiency virus type 1-infected cells to fas killing. AB - The importance of the Fas death pathway in human immunodeficiency virus (HIV) infection has been the subject of many studies. Missing from these studies is direct measurement of infected cell susceptibility to Fas-induced death. To address this question, we investigated whether T cells infected with HIV are more susceptible to Fas-induced death. We found that Fas cross-linking caused a decrease in the number of HIV-infected Jurkat T cells and CD4(+) peripheral blood leukocytes (PBLs). We confirmed this finding by demonstrating that there were more apoptotic infected than uninfected cells after Fas ligation. The increase in sensitivity of HIV-infected cells to Fas killing mapped to vpu, while nef, vif, vpr, and second exon of tat did not appear to contribute. Furthermore, expression of Vpu in Jurkat T cells rendered them more susceptible to Fas-induced death. These results show that HIV-infected cells are more sensitive to Fas-induced death and that the Vpu protein of HIV contributes to this sensitivity. The increased sensitivity of HIV-infected cells to Fas-induced death might help explain why these cells have such a short in vivo half-life. PMID- 9847312 TI - Genetic dissociation of the encapsidation and reverse transcription functions in the 5' R region of human immunodeficiency virus type 1. AB - The efficient packaging of genomic RNA into virions of human immunodeficiency virus type 1 (HIV-1) is directed by cis-acting encapsidation signals, which have been mapped to particular RNA stem-loop structures near the 5' end of the genome. Earlier studies have shown that three such stem-loops, located adjacent to the major 5' splice donor, are required for optimal packaging; more recent reports further suggest a requirement for the TAR and poly(A) hairpins of the 5' R region. In the present study, we have compared the phenotypes that result from mutating these latter elements in the HIV-1 provirus. Using a single-round infectivity assay, we find that mutations which disrupt base pairing in either the TAR or poly(A) stems cause profound defects in both packaging and viral replication. Decreased genomic packaging in a given mutant was always accompanied by increased packaging of spliced viral RNAs. Compensatory mutations that restored base pairing also restored encapsidation, indicating that the secondary structures of the TAR and poly(A) stems, rather than their primary sequences, are important for packaging activity. Despite having normal RNA contents, however, viruses with compensatory mutations at the base of the TAR stem were severely replication defective, owing to a defect in proviral DNA synthesis. Our findings thus confirm that the HIV-1 TAR stem-loop is required for at least three essential viral functions (transcriptional activation, RNA packaging, and reverse transcription) and reveal that its packaging and reverse transcription activities can be dissociated genetically by mutations at the base of the TAR stem. PMID- 9847313 TI - Colocalization of baculovirus IE-1 and two DNA-binding proteins, DBP and LEF-3, to viral replication factories. AB - We have recently identified a DNA-binding protein (DBP) from the baculovirus Bombyx mori nucleopolyhedrovirus (BmNPV) which can destabilize double-stranded DNA (V. S. Mikhailov, A. L. Mikhailova, M. Iwanaga, S. Gomi, and S. Maeda, J. Virol. 72:3107-3116, 1998). DBP was found to be an early gene product that was not present in budded or occlusion-derived virions. In order to characterize the localization of DBP during viral replication, BmNPV-infected BmN cells were examined by immunostaining and confocal microscopy with DBP antibodies. DBP first appeared as diffuse nuclear staining at 4 to 6 h postinfection (p.i.) and then localized to several specific foci within the nucleus at 6 to 8 h p.i. After the onset of viral DNA replication at around 8 h p.i., these foci began to enlarge and eventually occupied more than half of the nucleus by 14 h p.i. After the termination of viral DNA replication at about 20 h p.i., the DBP-stained regions appeared to break down into approximately 100 small foci within the nucleus. At 8 h p.i., the distribution of DBP as well as that of IE-1 or LEF-3 (two proteins involved in baculovirus DNA replication) overlapped well with that of DNA replication sites labeled with bromodeoxyuridine incorporation. Double-staining experiments with IE-1 and DBP or IE-1 and LEF-3 further confirmed that, between 8 and 14 h p.i., the distribution of IE-1 and LEF-3 overlapped with that of DBP. However, IE-1 localized to the specific foci prior to DBP or LEF-3 at 4 h p.i. In the presence of aphidicolin, an inhibitor of DNA synthesis, immature foci containing IE-1, LEF-3, and DBP were observed by 8 h p.i. However, the subsequent enlargement of these foci was completely suppressed, suggesting that the enlargement depended upon viral DNA replication. At 4 h p.i., the number of IE-1 foci correlated with the multiplicity of infection (MOI) between 0.4 and 10. At higher MOIs (e.g., 50), the number of foci plateaued at around 15. These results suggested that there are about 15 preexisting sites per nucleus which are associated with the initiation of viral DNA replication and assembly of viral DNA replication factories. PMID- 9847314 TI - Nucleoporins nup98 and nup214 participate in nuclear export of human immunodeficiency virus type 1 Rev. AB - Human immunodeficiency virus type 1 (HIV-1) Rev contains a leucine-rich nuclear export signal that is essential for its nucleocytoplasmic export mediated by hCRM1. We examined the role of selected nucleoporins, which are located in peripheral structures of the nuclear pore complex and are thought to be involved in export, in Rev function in human cells. First, we found that upon actinomycin D treatment, Nup98, but not Nup214 or Nup153, is able to translocate to the cytoplasm of HeLa cells, demonstrating that Nup98 may act as a soluble factor. We further showed that Rev can recruit Nup98 and Nup214, but not Nup153, to the nucleolus. We also found that the isolated FG-containing repeat domains of Nup98 and Nup214, but not those of Nup153, competitively inhibit the Rev/RRE-mediated expression of HIV. Taken together, the recruitment of Nup98 and Nup214 by Rev and the competitive inhibition exhibited by their NP domains demonstrate direct participation of Nup98 and Nup214 in the Rev-hCRM1-mediated export. PMID- 9847315 TI - Persistent Hz-1 virus infection in insect cells: evidence for insertion of viral DNA into host chromosomes and viral infection in a latent status. AB - Persistent/latent viral infections of insect cells are a prominent though poorly understood phenomenon. In this study, the long-term association between the Hz-1 virus and insect host cells, conventionally referred to as persistent viral infection, is described. With the aid of a newly developed fluorescent cell labeling system, we found that productive viral replication occurs by spontaneous viral reactivation in fewer than 0.2% of persistently infected cell lines over a 5-day period. Once viral reactivation takes place, the host cell dies. The persistently infected cells contain various amounts of viral DNA, and, in an extreme case, up to 16% of the total DNA isolated from infected cells could be of viral origin. Both pulsed-field gel electrophoresis and in situ hybridization experiments showed that some of these viral DNA molecules are inserted into the host chromosomes but that the rest of viral DNA copies are free from host chromosomes. Thus, Hz-1 virus is the first nonretroviral insect virus known to insert its genome into the host chromosome during the infection process. These data also suggest that the previously described persistent infection of Hz-1 virus in insect cells should be more accurately referred to as latent viral infection. PMID- 9847316 TI - Inhibition of influenza A virus replication by compounds interfering with the fusogenic function of the viral hemagglutinin. AB - Several compounds that specifically inhibited replication of the H1 and H2 subtypes of influenza virus type A were identified by screening a chemical library for antiviral activity. In single-cycle infections, the compounds inhibited virus-specific protein synthesis when added before or immediately after infection but were ineffective when added 30 min later, suggesting that an uncoating step was blocked. Sequencing of hemagglutinin (HA) genes of several independent mutant viruses resistant to the compounds revealed single amino acid changes that clustered in the stem region of the HA trimer in and near the HA2 fusion peptide. One of the compounds, an N-substituted piperidine, could be docked in a pocket in this region by computer-assisted molecular modeling. This compound blocked the fusogenic activity of HA, as evidenced by its inhibition of low-pH-induced cell-cell fusion in infected cell monolayers. An analog which was more effective than the parent compound in inhibiting virus replication was synthesized. It was also more effective in blocking other manifestations of the low-pH-induced conformational change in HA, including virus inactivation, virus induced hemolysis of erythrocytes, and susceptibility of the HA to proteolytic degradation. Both compounds inhibited viral protein synthesis and replication more effectively in cells infected with a virus mutated in its M2 protein than with wild-type virus. The possible functional relationship between M2 and HA suggested by these results is discussed. PMID- 9847317 TI - Full-length human immunodeficiency virus type 1 genomes from subtype C-infected seroconverters in India, with evidence of intersubtype recombination. AB - The development of an effective human immunodeficiency virus type 1 (HIV-1) vaccine is likely to depend on knowledge of circulating variants of genes other than the commonly sequenced gag and env genes. In addition, full-genome data are particularly limited for HIV-1 subtype C, currently the most commonly transmitted subtype in India and worldwide. Likewise, little is known about sequence variation of HIV-1 in India, the country facing the largest burden of HIV worldwide. Therefore, the objective of this study was to clone and characterize the complete genome of HIV-1 from seroconverters infected with subtype C variants in India. Cocultured HIV-1 isolates were obtained from six seroincident individuals from Pune, India, and virtually full-length HIV-1 genomes were amplified, cloned, and sequenced from each. Sequence analysis revealed that five of the six genomes were of subtype C, while one was a mosaic of subtypes A and C, with multiple breakpoints in env, nef, and the 3' long terminal repeat as determined by both maximal chi2 analysis and phylogenetic bootstrapping. Sequences were compared for preservation of known cytotoxic T lymphocyte (CTL) epitopes. Compared with those of the HIV-1LAI sequence, 38% of well-defined CTL epitopes were identical. The proportion of nonconservative substitutions for Env, at 61%, was higher (P < 0.001) than those for Gag (24%), Pol (18%), and Nef (32%). Therefore, characterized CTL epitopes demonstrated substantial differences from subtype B laboratory strains, which were most pronounced in Env. Because these clones were obtained from Indian seroconverters, they are likely to facilitate vaccine-related efforts in India by providing potential antigens for vaccine candidates as well as for assays of vaccine responsiveness. PMID- 9847318 TI - Formation of adeno-associated virus circular genomes is differentially regulated by adenovirus E4 ORF6 and E2a gene expression. AB - A central feature of the adeno-associated virus (AAV) latent life cycle is persistence in the form of both integrated and episomal genomes. However, the molecular processes associated with episomal long-term persistence of AAV genomes are only poorly understood. To investigate these mechanisms, we have utilized a recombinant AAV (rAAV) shuttle vector to identify circular AAV intermediates from transduced HeLa cells and primary fibroblasts. The unique structural features exhibited by these transduction intermediates included circularized monomer and dimer virus genomes in a head-to-tail array, with associated specific base pair alterations in the 5' viral D sequence. In HeLa cells, the abundance and stability of AAV circular intermediates were augmented by adenovirus expressing the E2a gene product. In the absence of E2a, adenovirus expressing the E4 open reading frame 6 gene product decreased the abundance of AAV circular intermediates, favoring instead the linear replication form monomer (Rfm) and dimer (Rfd) structures. In summary, the formation of AAV circular intermediates appears to represent a new pathway for AAV genome conversion, which is consistent with the head-to-tail concatemerization associated with latent-phase persistence of rAAV. A better understanding of this pathway may increase the utility of rAAV vectors for gene therapy. PMID- 9847319 TI - Diverse gene junctions of respiratory syncytial virus modulate the efficiency of transcription termination and respond differently to M2-mediated antitermination. AB - The ability of the diverse gene junctions of respiratory syncytial (RS) virus to signal the termination of transcription was analyzed. Nine dicistronic subgenomic replicons of RS virus were constructed; each contained one of the RS virus gene junctions in its natural upstream and downstream sequence context. The RNA synthesis activities of these subgenomic replicons were analyzed in the absence and presence of the M2 protein, which we showed previously to function as a transcription antiterminator. Our data showed that the efficiency with which the polymerase terminated transcription was affected by the gene junction that it encountered. The M2 protein significantly decreased the efficiency of the termination of transcription, resulting in increased levels of readthrough transcription at all the gene junctions. The diverse gene junctions fell into three broad groups with respect to their ability to signal transcription termination. One group of gene junctions (NS1/NS2, NS2/N, M2/L, and L/trailer) showed inefficient termination in the absence or the presence of the M2 protein. A second group of gene junctions (N/P, P/M, M/SH, SH/G, and G/F) terminated transcription efficiently. The SH/G gene junction terminated transcription with the greatest efficiency and produced low levels of readthrough transcripts in the absence or the presence of the M2 protein, correlating with the absence of SH/G polycistronic transcripts in RS virus-infected cells. The F/M2 gene junction was particularly sensitive to the M2 protein: it efficiently signaled termination in the absence of the M2 protein but produced high levels of readthrough transcripts in the presence of the M2 protein. This result suggests that the M2 protein may regulate its own production by negative feedback. The data presented here show that the different gene junctions of RS virus do modulate RS virus transcription termination. The M2 protein reduced termination at all gene junctions. The magnitude of antitermination due to the M2 protein, however, varied at the different gene junctions. The data presented here indicate that the mechanism for the regulation of RS virus gene expression is more complex than was previously appreciated. PMID- 9847320 TI - Processing of the human coronavirus 229E replicase polyproteins by the virus encoded 3C-like proteinase: identification of proteolytic products and cleavage sites common to pp1a and pp1ab. AB - Replicase gene expression by the human coronavirus 229E involves the synthesis of two large polyproteins, pp1a and pp1ab. Experimental evidence suggests that these precursor molecules are subject to extensive proteolytic processing. In this study, we show that a chymotrypsin-like enzyme, the virus-encoded 3C-like proteinase (3CLpro), cleaves within a common region of pp1a and pp1ab (amino acids 3490 to 4068) at four sites. trans-cleavage assays revealed that polypeptides of 5, 23, 12, and 16 kDa are processed from pp1a/pp1ab by proteolysis of the peptide bonds Q3546/S3547, Q3629/S3630, Q3824/N3825, and Q3933/A3934. Relative rate constants for the 3CLpro-mediated cleavages Q2965/A2966, Q3267/S3268, Q3824/N3825, and Q3933/A3934 were derived by competition experiments using synthetic peptides and recombinant 3CLpro. The results indicate that coronavirus cleavage sites differ significantly with regard to their susceptibilities to proteolysis by 3CLpro. Finally, immunoprecipitation with specific rabbit antisera was used to detect the pp1a/pp1ab processing end products in virus-infected cells, and immunofluorescence data that suggest an association of these polypeptides with intracellular membranes were obtained. PMID- 9847321 TI - Effects of maternal antibodies on protection and development of antibody responses to human rotavirus in gnotobiotic pigs. AB - Although maternal antibodies can protect against infectious disease in infancy, they can also suppress active immune responses. The effects of circulating maternal antibodies, with and without colostrum and milk antibodies, on passive protection and active immunity to human rotavirus (HRV) were examined in gnotobiotic pigs. Pigs received intraperitoneal injections of high-titer serum (immune pigs [groups 1 and 2]) from immunized sows, low-titer serum from naturally infected sows (control pigs [groups 3 and 4]), or no serum (group 5). Immune or control colostrum and milk were added to the diet of groups 2 and 4, respectively. After inoculation (3 to 5 days of age) and challenge (postinoculation day [PID] 21) with virulent HRV, the effects of maternal antibodies on protection (from diarrhea and virus shedding), and on active antibody responses (measured by quantitation of antibody-secreting cells [ASC] in intestinal and systemic lymphoid tissues by ELISPOT) were evaluated. Groups 1 and 2 had significantly less diarrhea and virus shedding after inoculation but higher rates of diarrhea and virus shedding after challenge than did groups 3 and 5. Group 1 and 2 pigs had significantly fewer immunoglobulin A (IgA) ASC in intestinal tissues at PID 21 and at postchallenge day (PCD) 7 compared to group 5. Significantly fewer IgG ASC were present in the intestines of group 2 pigs at PID 21 and PCD 7 compared to group 5. There was a trend towards fewer ASC in intestinal tissues of group 2 than group 1, from PID 21 on, with significantly fewer IgA ASC at PCD 7. IgG ASC in the duodenum and mesenteric lymph nodes of group 3 and 4 pigs were significantly fewer than in group 5 at PCD 7. These decreases in ASC emphasize the role of passive antibodies in impairing induction of ASC rather than in merely suppressing the function of differentiated B cells. To be successful, vaccines intended for populations with high titers of maternal antibodies (infants in developing countries) may require higher titers of virus, multiple doses, or improved delivery systems, such as the use of microencapsulation or immune stimulating complexes, to overcome the suppressive effects of maternal antibodies. PMID- 9847322 TI - Spatial perturbations within an RNA promoter specifically recognized by a viral RNA-dependent RNA polymerase (RdRp) reveal that RdRp can adjust its promoter binding sites. AB - RNA synthesis during viral replication requires specific recognition of RNA promoters by the viral RNA-dependent RNA polymerase (RdRp). Four nucleotides ( 17, -14, -13, and -11) within the brome mosaic virus (BMV) subgenomic core promoter are required for RNA synthesis by the BMV RdRp (R. W. Siegel et al., Proc. Natl. Acad. Sci. USA 94:11238-11243, 1997). The spatial requirements for these four nucleotides and the initiation (+1) cytidylate were examined in RNAs containing nucleotide insertions and deletions within the BMV subgenomic core promoter. Spatial perturbations between nucleotides -17 and -11 resulted in decreased RNA synthesis in vitro. However, synthesis was still dependent on the key nucleotides identified in the wild-type core promoter and the initiation cytidylate. In contrast, changes between nucleotides -11 and +1 had a less severe effect on RNA synthesis but resulted in RNA products initiated at alternative locations in addition to the +1 cytidylate. The results suggest a degree of flexibility in the recognition of the subgenomic promoter by the BMV RdRp and are compared with functional regions in other DNA and RNA promoters. PMID- 9847323 TI - Microglia express CCR5, CXCR4, and CCR3, but of these, CCR5 is the principal coreceptor for human immunodeficiency virus type 1 dementia isolates. AB - Microglia are the main human immunodeficiency virus (HIV) reservoir in the central nervous system and most likely play a major role in the development of HIV dementia (HIVD). To characterize human adult microglial chemokine receptors, we analyzed the expression and calcium signaling of CCR5, CCR3, and CXCR4 and their roles in HIV entry. Microglia expressed higher levels of CCR5 than of either CCR3 or CXCR4. Of these three chemokine receptors, only CCR5 and CXCR4 were able to transduce a signal in microglia in response to their respective ligands, MIP-1beta and SDF-1alpha, as recorded by single-cell calcium flux experiments. We also found that CCR5 is the predominant coreceptor used for infection of human adult microglia by the HIV type 1 dementia isolates HIV-1DS br, HIV-1RC-br, and HIV-1YU-2, since the anti-CCR5 antibody 2D7 was able to dramatically inhibit microglial infection by both wild-type and single-round luciferase pseudotype reporter viruses. Anti-CCR3 (7B11) and anti-CXCR4 (12G5) antibodies had little or no effect on infection. Last, we found that virus pseudotyped with the DS-br and RC-br envelopes can infect cells transfected with CD4 in conjunction with the G-protein-coupled receptors APJ, CCR8, and GPR15, which have been previously implicated in HIV entry. PMID- 9847325 TI - Chimeric lyssavirus glycoproteins with increased immunological potential. AB - The rabies virus glycoprotein molecule (G) can be divided into two parts separated by a flexible hinge: the NH2 half (site II part) containing antigenic site II up to the linear region (amino acids [aa] 253 to 275 encompassing epitope VI [aa 264]) and the COOH half (site III part) containing antigenic site III and the transmembrane and cytoplasmic domains. The structural and immunological roles of each part were investigated by cell transfection and mouse DNA-based immunization with homogeneous and chimeric G genes formed by fusion of the site II part of one genotype (GT) with the site III part of the same or another GT. Various site II-site III combinations between G genes of PV (Pasteur virus strain) rabies (GT1), Mokola (GT3), and EBL1 (European bat lyssavirus 1 [GT5]) viruses were tested. Plasmids pGPV-PV, pGMok-Mok, pGMok-PV, and pGEBL1-PV induced transient expression of correctly transported and folded antigens in neuroblastoma cells and virus-neutralizing antibodies against parental viruses in mice, whereas, pG-PVIII (site III part only) and pGPV-Mok did not. The site III part of PV (GT1) was a strong inducer of T helper cells and was very effective at presenting the site II part of various GTs. Both parts are required for correct folding and transport of chimeric G proteins which have a strong potential value for immunological studies and development of multivalent vaccines. Chimeric plasmid pGEBL1-PV broadens the spectrum of protection against European lyssavirus genotypes (GT1, GT5, and GT6). PMID- 9847324 TI - Transactivation of a ribosomal gene by simian virus 40 large-T antigen requires at least three activities of the protein. AB - Simian virus 40 large-T antigen transactivates the ribosomal genes which are transcribed by RNA polymerase (pol I), as well as genes that are dependent on either pol II or pol III. This report identifies regions and activities of T antigen that are required to transactivate a pol I-dependent rat ribosomal gene promoter. By using the rat ribosomal gene (rDNA) promoter linked to a chloramphenicol acetyltransferase gene, we show that at least three separable T antigen regions are necessary to achieve wild-type levels of transactivation of rDNA in transiently transfected monkey cells. One activity depends on the region of T antigen shared with small-t antigen (T/t common region). A second activity maps to amino acids 109 to 626 and is highly sensitive to mutational inactivation. Complementation analyses suggest that at least one activity in this region is independent of and must be in cis with the activity within the T/t common region. In addition, a functional nuclear localization signal is required for maximal T-antigen-mediated transactivation of rat rDNA. The three activities work in concert to override cellular species-specific controls and transactivate the rat ribosomal gene promoter. Finally, we provide evidence that although the tumor suppressor protein Rb has been shown to repress a pol I-dependent promoter, transactivation of the rat rDNA promoter does not depend on T antigen's ability to bind the tumor suppressor product Rb. PMID- 9847326 TI - Sequence-specific and/or stereospecific constraints of the U3 enhancer elements of MCF 247-W are important for pathogenicity. AB - The oncogenic potential of many nonacute retroviruses is dependent on the duplication of the enhancer sequences present in the unique 3' (U3) region of the long terminal repeat (LTR). In a molecular clone (MCF 247-W) of the murine leukemia virus MCF 247, a leukemogenic mink cell focus-inducing (MCF) virus, the U3 enhancer sequences are tandemly repeated in the LTR. We mutated the enhancer region of MCF 247-W to test the hypothesis that the duplicated enhancer sequences of this virus have a sequence-specific and/or a stereospecific role in enhancer function required for transformation. In one virus, we inserted 14 nucleotide bp into the novel sequence generated at the junction of the two enhancers to generate an MCF virus with an interrupted enhancer region. In the second virus, only one copy of the enhancer sequences was present. This second virus also lacked the junction sequence present between the two enhancers of MCF 247-W. Both viruses were less leukemogenic and had a longer mean latency period than MCF 247 W. These data indicate that the sequence generated at the junction of the two enhancers and/or the stereospecific arrangement of the two enhancer elements are required for the full oncogenic potential of MCF 247-W. We analyzed proviral LTRs within the c-myc locus in tumor DNAs from mice injected with the MCF virus with the interrupted enhancer region. Some of the proviral LTRs integrated upstream of c-myc contain enhancer regions that are larger than those of the injected virus. These results are consistent with the suggestion that the virus with an interrupted enhancer changes in vivo to perform its role in the transformation of T cells. PMID- 9847327 TI - Matrix protein of rabies virus is responsible for the assembly and budding of bullet-shaped particles and interacts with the transmembrane spike glycoprotein G. AB - To elucidate the functions of rhabdovirus matrix (M) protein, we determined the localization of M in rabies virus (RV) and analyzed the properties of an M deficient RV mutant. We provide evidence that M completely covers the ribonucleoprotein (RNP) coil and keeps it in a condensed form. As determined by cosedimentation experiments, not only the M-RNP complex but also M alone was found to interact specifically with the glycoprotein G. In contrast, an interaction of G with the nucleoprotein N or M-less RNP was not observed. In the absence of M, infectious particles were mainly cell associated and the yield of cell-free infectious virus was reduced by as much as 500,000-fold, demonstrating the crucial role of M in virus budding. Supernatants from cells infected with the M-deficient RV did not contain the typical bullet-shaped rhabdovirus particles but instead contained long, rod-shaped virions, demonstrating severe impairment of the virus formation process. Complementation with M protein expressed from plasmids rescued rhabdovirus formation. These results demonstrate the pivotal role of M protein in condensing and targeting the RNP to the plasma membrane as well as in incorporation of G protein into budding virions. PMID- 9847328 TI - Generation of bovine respiratory syncytial virus (BRSV) from cDNA: BRSV NS2 is not essential for virus replication in tissue culture, and the human RSV leader region acts as a functional BRSV genome promoter. AB - In order to generate recombinant bovine respiratory syncytial virus (BRSV), the genome of BRSV strain A51908, variant ATue51908, was cloned as cDNA. We provide here the sequence of the BRSV genome ends and of the entire L gene. This completes the sequence of the BRSV genome, which comprises a total of 15,140 nucleotides. To establish a vaccinia virus-free recovery system, a BHK-derived cell line stably expressing T7 RNA polymerase was generated (BSR T7/5). Recombinant BRSV was reproducibly recovered from cDNA constructs after T7 RNA polymerase-driven expression of antigenome sense RNA and of BRSV N, P, M2, and L proteins from transfected plasmids. Chimeric viruses in which the BRSV leader region was replaced by the human respiratory syncytial virus (HRSV) leader region replicated in cell culture as efficiently as their nonchimeric counterparts, demonstrating that all cis-acting sequences of the HRSV promoter are faithfully recognized by the BRSV polymerase complex. In addition, we report the successful recovery of a BRSV mutant lacking the complete NS2 gene, which encodes a nonstructural protein of unknown function. The NS2-deficient BRSV replicated autonomously and could be passaged, demonstrating that NS2 is not essential for virus replication in cell culture. However, growth of the mutant was considerably slower than and final infectious titers were reduced by a factor of at least 10 compared to wild-type BRSV, indicating that NS2 provides a supporting factor required for full replication capacity. PMID- 9847329 TI - The adeno-associated virus type 2 regulatory proteins rep78 and rep68 interact with the transcriptional coactivator PC4. AB - The adeno-associated virus type 2 (AAV-2) Rep78/Rep68 regulatory proteins are pleiotropic effectors of viral and cellular DNA replication, of cellular transformation by viral and cellular oncogenes, and of homologous and heterologous gene expression. To search for cellular proteins involved in mediating these functions, we used Rep68 as bait in the yeast two-hybrid system and identified the transcriptional coactivator PC4 as a Rep interaction partner. PC4 has been shown to mediate transcriptional activation by a variety of sequence specific transcription factors in vitro. Rep amino acids 172 to 530 were sufficient and amino acids 172 to 224 were absolutely necessary for the interaction with PC4. The PC4 domains required for interaction were mapped to the C-terminal single-stranded DNA-binding domain of PC4. In glutathione S transferase (GST) pull-down assays, in vitro-transcribed and -translated Rep78 or Rep68 proteins were bound specifically by GST-PC4 fusion proteins. Similarly, PC4 expressed in Escherichia coli was bound by GST-Rep fusion proteins, confirming the direct interaction between Rep and PC4 in vitro. Rep was found to have a higher affinity for the nonphosphorylated, transcriptionally active form of PC4 than for the phosphorylated, transcriptionally inactive form. The latter is predominant in nuclear extracts of HeLa or 293 cells. In the yeast system, but not in vitro, Rep-PC4 interaction was disrupted by a point mutation in the putative nucleotide-binding site of Rep68, suggesting that a stable interaction between Rep and PC4 in vivo is ATP dependent. This mutation has also been shown to impair Rep function in AAV-2 DNA replication and in inhibition of gene expression and inducible DNA amplification. Cytomegalovirus promoter-driven overexpression of PC4 led to transient accumulation of nonphosphorylated PC4 with concomitant downregulation of all three AAV-2 promoters in the absence of helper virus. In the presence of adenovirus, this effect was relieved. These results imply an involvement of the transcriptional coactivator PC4 in the regulation of AAV-2 gene expression in the absence of helper virus. PMID- 9847330 TI - Porcine reproductive and respiratory syndrome virus comparison: divergent evolution on two continents. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) is a recently described arterivirus responsible for disease in swine worldwide. Comparative sequence analysis of 3'-terminal structural genes of the single-stranded RNA viral genome revealed the presence of two genotypic classes of PRRSV, represented by the prototype North American and European strains, VR-2332 and Lelystad virus (LV), respectively. To better understand the evolution and pathogenicity of PRRSV, we obtained the 12,066-base 5'-terminal nucleotide sequence of VR-2332, encoding the viral replication activities, and compared it to those of LV and other arteriviruses. VR-2332 and LV differ markedly in the 5' leader and sections of the open reading frame (ORF) 1a region. The ORF 1b sequence was nearly colinear but varied in similarity of proteins encoded in identified regions. Furthermore, molecular and biochemical analysis of subgenomic mRNA (sgmRNA) processing revealed extensive variation in the number of sgmRNAs which may be generated during infection and in the lengths of noncoding sequence between leader-body junctions and the translation-initiating codon AUG. In addition, VR 2332 and LV select different leader-body junction sites from a pool of similar candidate sites to produce sgmRNA 7, encoding the viral nucleocapsid protein. The presence of substantial variations across the entire genome and in sgmRNA processing indicates that PRRSV has evolved independently on separate continents. The near-simultaneous global emergence of a new swine disease caused by divergently evolved viruses suggests that changes in swine husbandry and management may have contributed to the emergence of PRRS. PMID- 9847331 TI - Immunization of woodchucks with plasmids expressing woodchuck hepatitis virus (WHV) core antigen and surface antigen suppresses WHV infection. AB - DNA vaccination can induce humoral and cellular immune response to viral antigens and confer protection to virus infection. In woodchucks, we tested the protective efficacy of immune response to woodchuck hepatitis core antigen (WHcAg) and surface antigen (WHsAg) of woodchuck hepatitis virus (WHV) elicited by DNA-based vaccination. Plasmids pWHcIm and pWHsIm containing WHV c- or pre-s2/s genes expressed WHcAg and WHsAg in transient transfection assays. Pilot experiments in mice revealed that a single intramuscular injection of 100 microgram of plasmid pWHcIm DNA induced an anti-WHcAg titer over 1:300 that was enhanced by boost injections. However, two injections of 100 microgram of pWHcIm did not induce detectable anti-WHcAg in woodchucks. With an increase in the dose to 1 mg of pWHcIm per injection, transient anti-WHcAg response and WHcAg-specific proliferation of peripheral mononuclear blood cells (PMBCs) appeared in woodchucks after repeated immunizations. Four woodchucks vaccinated with pWHcIm were challenged with 10(4) or 10(5) of the WHV 50% infective dose. They remained negative for markers of WHV replication (WHV DNA and WHsAg) in peripheral blood and developed anti-WHs in week 5 after challenge. In contrast, woodchucks not immunized or immunized with the control vector pcDNA3 developed acute WHV infection. Two woodchucks immunized with 1 mg of pWHsIm developed WHsAg-specific proliferative response of PBMCs but no measurable anti-WHsAg response. A rapid anti-WHsAg response developed during week 2 after virus challenge. Neither woodchuck developed any signs of WHV infection. These data indicate that DNA based vaccination with WHcAg and WHsAg can elicit immunity to WHV infection. PMID- 9847332 TI - RNase L inhibitor is induced during human immunodeficiency virus type 1 infection and down regulates the 2-5A/RNase L pathway in human T cells. AB - The interferon-regulated 2-5A/RNase L pathway plays a major role in the antiviral and antiproliferative activities of these cytokines. Several viruses, however, have evolved strategies to escape the antiviral activity of the 2-5A/RNase L pathway. In this context, we have cloned a cDNA coding for the RNase L inhibitor (RLI), a protein that specifically inhibits RNase L and whose regulated expression in picornavirus-infected cells down regulates the activity of the 2 5A/RNase L pathway. We show here that RLI increases during the course of human immunodeficiency virus type 1 (HIV-1) infection, which may be related to the downregulation of RNase L activity that has been described to occur in HIV infected cells. In order to establish a possible causal relationship between these observations, we have stably transfected H9 cells with RLI sense or antisense cDNA-expressing vectors. The overexpression of RLI causes a decrease in RNase L activity and a twofold enhancement of HIV production. This increase in HIV replication correlates with an increase in HIV RNA and proteins. In contrast, reduction of RLI levels in RLI antisense cDNA-expressing clones reverses the inhibition of RNase L activity associated with HIV multiplication and leads to a threefold decrease in the viral load. This anti-HIV activity correlated with a decrease in HIV RNA and proteins. These findings demonstrate that the level of RLI, via its modulation of RNase L activity, can severely impair HIV replication and suggest the involvement of RLI in the inhibition of the 2-5A/RNase L system observed during HIV infection. PMID- 9847333 TI - Regulation of RNA synthesis by the genomic termini of vesicular stomatitis virus: identification of distinct sequences essential for transcription but not replication. AB - The RNA-dependent RNA polymerase of vesicular stomatitis virus (VSV), a nonsegmented negative-strand RNA virus, directs two discrete RNA synthetic processes, transcription and replication. Available evidence suggests that the two short extragenic regions at the genomic termini, the 3' leader (Le) and the complement of the 5' trailer (TrC), contain essential signals for these processes. We examined the roles in transcription and replication of sequences in Le and TrC by monitoring the effects of alterations to the termini of subgenomic replicons, or infectious viruses, on these RNA synthetic processes. Distinct elements in Le were found to be required for transcription that were not required for replication. The promoter for mRNA transcription was shown to include specific sequence elements within Le at positions 19 to 29 and 34 to 46, a separate element at nucleotides 47 to 50, the nontranscribed leader-N gene junction. The sequence requirements for transcription within the Le region could not be supplied by sequences found at the equivalent positions in TrC. In contrast, sequences from either Le or TrC functioned well to signal replication, indicating that within the confines of the VSV termini, the sequence requirements for replication were less stringent. Deletions engineered at the termini showed that the terminal 15 nucleotides of either Le or TrC allowed a minimal level of replication. Within these confines, levels of replication were affected by both the extent of complementarity between the genomic termini and the involvement of the template in transcription. In agreement with our previous observations, increasing the extent of complementarity between the natural termini increased levels of replication, and this effect was most operative at the extreme genome ends. In addition, abolishing the use of Le as a promoter for transcription enhanced replication. These analyses (i) identified signals at the termini required for transcription and replication and (ii) showed that Le functions as a less efficient promoter for replication than TrC at least in part because of its essential role in transcription. Consequently, these observations help explain the asymmetry of VSV replication which results in the synthesis of more negative- than positive-sense replication products in infected cells. PMID- 9847334 TI - The 5' terminal trailer region of vesicular stomatitis virus contains a position dependent cis-acting signal for assembly of RNA into infectious particles. AB - The cis-acting genomic RNA requirements for the assembly of vesicular stomatitis virus (VSV) ribonucleocapsids into infectious particles were investigated. Using a biological assay based on particle infectivity, we demonstrated that subgenomic replicons that contained all four possible combinations of the natural genomic termini, the 3' leader (Le) and 5' trailer (Tr) regions, were replication competent; however, a 3' copyback replicon (3'CB), containing the natural 3' terminus but having the 5' Tr replaced by a sequence complementary to the 3' Le for 46 nucleotides, was unable to assemble infectious particles, despite efficient replication. When a copy of Tr was inserted 51 nucleotides from the 5' end of 3'CB, infectious particles were produced. However, analysis of the replication products of these particles showed that the 51 nucleotides which corresponded to the Le complement sequences at the 5' terminus were removed during RNA replication, thus restoring the wild-type 5' Tr to the exact 5' terminus. These data showed that a cis-acting signal was necessary for assembly of VSV RNAs into infectious particles and that this signal was supplied by Tr when located at the 5' end. The regions within Tr required for assembly were analyzed by a series of deletions and exchanges for Le complement sequences, which demonstrated that the 5' terminal 29 nucleotides of Tr allowed assembly of infectious particles but that the 5' terminal 22 nucleotides functioned poorly. Deletions in Tr also altered the balance between negative- and positive-strand genomic RNA and affected levels of replication. RNAs that retained fewer than 45 but at least 22 nucleotides of the 5' terminus could replicate but were impaired in RNA replication, and RNAs that retained only 14 nucleotides of the 5' terminus were severely reduced in ability to replicate. These data define the VSV Tr as a position-dependent, cis-acting element for the assembly of RNAs into infectious particles, and they delineate RNA sequences that are essential for negative strand RNA synthesis. These observations are consistent with, and offer an explanation for, the absence of 3' copyback defective interfering particles in nature. PMID- 9847336 TI - Trafficking to the plasma membrane of the seven-transmembrane protein encoded by human herpesvirus 6 U51 gene involves a cell-specific function present in T lymphocytes. AB - The sequence of human herpesvirus 6 (HHV-6) U51 open reading frame predicts a protein of 301 amino acid residues with seven transmembrane domains. To identify and characterize U51, we derived antipeptide polyclonal antibodies and developed a transient expression assay. We ascertained that U51 was synthesized in cord blood mononuclear cells infected with either variant A- or variant B-HHV-6 and was transported to the surface of productively infected cells. When synthesized in transient expression systems, U51 intracellular trafficking was regulated in a cell-type-dependent fashion. In human monolayer HEK-293 and 143tk- cells, U51 accumulated predominantly in the endoplasmic reticulum and failed to be transported to the cell surface. In contrast, in T-lymphocytic cell lines J-Jhan, Molt-3, and Jurkat, U51 was successfully transported to the plasma membrane. We infer that transport of U51 to the cell surface requires a cell-specific function present in activated T lymphocytes and T-cell lines. PMID- 9847335 TI - Determinant in human immunodeficiency virus type 1 for efficient replication under cytokine-induced CD4(+) T-helper 1 (Th1)- and Th2-type conditions. AB - Cytokines are potent stimuli for CD4(+)-T-cell differentiation. Among them, interleukin-12 (IL-12) and IL-4 induce naive CD4(+) T cells to become T-helper 1 (Th1) or Th2 cells, respectively. In this study we found that macrophage-tropic human immunodeficiency virus type 1 (HIV-1) strains replicated more efficiently in IL-12-induced Th1-type cultures derived from normal CD4(+) T cells than did T cell-line-tropic (T-tropic) strains. In contrast, T-tropic strains preferentially infected IL-4-induced Th2-type cultures derived from the same donor CD4(+) T cells. Additional studies using chimeric viruses demonstrated that the V3 region of HIV-1 gp120 was the principal determinant for efficiency of replication. Cell fusion analysis showed that cells expressing envelope protein from a T-tropic strain effectively fused with IL-4-induced Th2-type culture cells. Flow cytometric analysis showed that the level of CCR5 expression was higher on IL-12 induced Th1-type culture cells, whereas CXCR4 was highly expressed on IL-4 induced Th2-type culture cells, although a low level of CXCR4 expression was observed on IL-12-induced Th1-type culture cells. These results indicate that HIV 1 isolates exhibit differences in the ability to infect CD4(+)-T-cell subsets such as Th1 or Th2 cells and that this difference may partly correlate with the expression of particular chemokine receptors on these cells. The findings suggest that immunological conditions are one of the factors responsible for inducing selection of HIV-1 strains. PMID- 9847337 TI - Accessing Epstein-Barr virus-specific T-cell memory with peptide-loaded dendritic cells. AB - The conventional means of studying Epstein-Barr virus (EBV)-induced cytotoxic T lymphocyte (CTL) memory, by in vitro stimulation with the latently infected autologous lymphoblastoid cell line (LCL), has important limitations. First, it gives no information on memory to lytic cycle antigens; second, it preferentially amplifies the dominant components of latent antigen-specific memory at the expense of key subdominant reactivities. Here we describe an alternative approach, based on in vitro stimulation with epitope peptide-loaded dendritic cells (DCs), which allows one to probe the CTL repertoire for any individual reactivity of choice; this method proved significantly more efficient than stimulation with peptide alone. Using this approach we first show that reactivities to the immunodominant and subdominant lytic cycle epitopes identified by T cells during primary EBV infection are regularly detectable in the CTL memory of virus carriers; this implies that in such carriers chronic virus replication remains under direct T-cell control. We further show that subdominant latent cycle reactivities to epitopes in the latent membrane protein LMP2, though rarely undetectable in LCL-stimulated populations, can be reactivated by DC stimulation and selectively expanded as polyclonal CTL lines; the adoptive transfer of such preparations may be of value in targeting certain EBV-positive malignancies. PMID- 9847338 TI - Two nucleotides immediately upstream of the essential A6G3 slippery sequence modulate the pattern of G insertions during Sendai virus mRNA editing. AB - Editing of paramyxovirus P gene mRNAs occurs cotranscriptionally and functions to fuse an alternate downstream open reading frame to the N-terminal half of the P protein. G residues are inserted into a short G run contained within a larger purine run (AnGn) in this process, by a mechanism whereby the transcribing polymerase stutters (i.e., reads the same template cytosine more than once). Although Sendai virus (SeV) and bovine parainfluenza virus type 3 (bPIV3) are closely related, the G insertions in their P mRNAs are distributed differently. SeV predominantly inserts a single G residue within the G run of the sequence 5' AACAAAAAAGGG, whereas bPIV3 inserts one to six G's at roughly equal frequency within the sequence 5' AUUAAAAAAGGGG (differences are underlined). We have examined how the cis-acting editing sequence determines the number of G's inserted, both in a transfected cell system using minigenome analogues and by generating recombinant viruses. We found that the presence of four rather than three G's in the purine run did not affect the distribution of G insertions. However, when the underlined AC of the SeV sequence was replaced by the UU found in bPIV3, the editing phenotype from both the minigenome and the recombinant virus resembled that found in natural bPIV3 infections (i.e., a significant fraction of the mRNAs contained two to six G insertions). The two nucleotides located just upstream of the polypurine tract are thus key determinants of the editing phenotype of these viruses. Moreover, the minimum number of A residues that will promote SeV editing phenotype is six but can be reduced to five when the upstream AC is replaced by UU. A model for how the upstream dinucleotide controls the insertion phenotype is presented. PMID- 9847339 TI - Diminished production of human immunodeficiency virus type 1 in astrocytes results from inefficient translation of gag, env, and nef mRNAs despite efficient expression of Tat and Rev. AB - Astrocytes infected with human immunodeficiency virus type 1 (HIV-1) produce only minimal quantities of virus. The molecular events that limit acute-phase HIV-1 infection of astrocytes were examined after inducing acute-phase replication by transfection with the pNL4-3 proviral plasmid. The levels of HIV-1 mRNA were similarly high in both astrocytes and HeLa cells, but astrocytes produced approximately 50-fold less supernatant p24 than HeLa cells. We found that diminished HIV-1 production in astrocytes resulted from inefficient translation of gag, env, and nef mRNAs that were efficiently transported to the cytoplasm. Tat- or Rev-dependent reporter constructs showed no defect in Tat or Rev function in astrocytes compared with HeLa cells. HIV-1 mRNAs were correctly spliced, but only Rev and Tat proteins were efficiently translated from their native mRNAs. Pulse-chase labelling and immunoblot experiments revealed no defect in protein processing, but levels of Gag, Env, or Nef protein expressed were dramatically reduced in astrocytes compared to HeLa cells. These results demonstrate that inefficient translation of HIV-1 structural proteins underlies the restricted infection of astrocytes. The efficient expression of functional Tat and Rev by astrocytes may contribute to HIV-1 neuropathogenesis. PMID- 9847340 TI - Dynamics of human immunodeficiency virus type 1 replication in vertically infected infants. AB - Plasma human immunodeficiency virus type 1 (HIV-1) turnover and kinetics were studied in children aged 15 days to 2 years following the initiation of a triple antiretroviral drug regimen consisting of zidovudine, lamivudine, and nevirapine. HIV-1 turnover was at least as rapid as that previously described in adults; turnover rates were more rapid in infants and children aged 3 months to 2 years than in infants less than 3 months of age. These data confirm the central role of HIV-1 replication in the pathogenesis of vertical HIV-1 infection and reinforce the importance of early, potent combination therapies for the long-term control of HIV-1 replication. PMID- 9847341 TI - Mammary gland expression of mouse mammary tumor virus is regulated by a novel element in the long terminal repeat. AB - Mouse mammary tumor virus (MMTV) infects both lymphoid tissue and lactating mammary gland during its infectious cycle, but some endogenous MMTVs are transcribed only in lymphoid cells. We found a lymphoid cell-specific endogenous MMTV that was converted to a milk-borne, infectious virus through recombination with an exogenously transmitted MMTV. The changed expression pattern correlated with the alteration of a single base pair in the long terminal repeat of the lymphoid cell-specific virus. Transgenic mice with the element from either the milk-borne or lymphoid cell-specific virus upstream of the chloramphenicol acetyltransferase reporter gene showed the same pattern of expression as the virus from which the regulatory sequences were derived. Electrophoretic mobility shift assays with mammary cell extracts showed that the site from the milk-borne virus was preferentially bound by a prolactin-inducible factor that poorly bound the altered site from the lymphoid cell-specific virus. The complex that formed on the milk-borne virus-specific oligonucleotide supershifted with anti-Stat5b antibody. Mice lacking either Stat5a or Stat5b had dramatically reduced levels of MMTV transcripts in mammary gland but not in lymphoid tissue. Thus, a member of the STAT family of transcription factors is involved in the tissue-specific expression of mouse mammary tumor virus in vivo. This is the first example of the involvement of a member of the STAT family of transcription factors in the control of tissue-specific expression. PMID- 9847343 TI - Model for polymerase access to the overlapped L gene of respiratory syncytial virus. AB - The last two genes of respiratory syncytial virus (RSV), M2 and L, overlap by 68 nucleotides, an arrangement which has counterparts in a number of nonsegmented negative-strand RNA viruses. Thus, the gene-end (GE) signal of M2 lies downstream of the L gene-start (GS) signal, separated by 45 nucleotides. Since RSV transcription ostensibly is sequential and unidirectional from a single promoter within the 3' leader region, it was unclear how the polymerase accesses the L GS signal. Furthermore, it was previously shown that 90% of transcripts which are initiated at the L GS signal are polyadenylated and terminated at the M2 GE signal, yielding a short, truncated L mRNA as the major transcription product of the L gene. Despite these apparent down-regulatory features, we show that the accumulation of full-length L mRNA during RSV infection is only sixfold less than that of its upstream neighbor, M2. We used cDNA-encoded genome analogs in an intracellular transcription assay to investigate the mechanism of transcription of the overlapped genes. Expression of L was found to be dependent on sequential transcription from the 3' end of the genome. Apart from the L GS signal, the only other strict requirement for initiation at L was the M2 GE signal. This implies that the polymerase accesses the L GS signal only following arrival at the M2 GE signal. Thus, polymerase which terminates at the M2 GE signal presumably scans upstream to initiate at the L GS signal. This also would provide a mechanism whereby polymerase which terminates prematurely during transcription of L could recycle from the M2 GE signal to the L GS signal, thereby accounting for the unexpectedly high level of synthesis of full-length L mRNA. The sequence and spacing between the two signals were not critical. Furthermore, the polymerase also was capable of efficiently transcribing from an L GS signal placed downstream of the M2 GE signal, implying that the overlapping arrangement is not obligatory. When copies of the L GS signal were placed concurrently upstream and downstream of the M2 GE signal, both were utilized. This finding indicates that a polymerase situated at a GE signal is capable of scanning for a GS signal in either the upstream or downstream direction and thereafter initiating transcription. PMID- 9847344 TI - Analysis of murine CD8(+) T-cell clones specific for the Dengue virus NS3 protein: flavivirus cross-reactivity and influence of infecting serotype. AB - Serotype-cross-reactive dengue virus-specific cytotoxic T lymphocytes (CTL) induced during a primary dengue virus infection are thought to play a role in the immunopathogenesis of dengue hemorrhagic fever (DHF) during a secondary dengue virus infection. Although there is no animal model of DHF, we previously reported that murine dengue virus-specific CTL responses are qualitatively similar to human dengue virus-specific CTL responses. We used BALB/c mice to study the specificity of the CTL response to an immunodominant epitope on the dengue virus NS3 protein. We mapped the minimal H-2Kd-restricted CTL epitope to residues 298 to 306 of the dengue type 2 virus NS3 protein. In short-term T-cell lines and clones, the predominant CD8(+) CTL to this epitope in mice immunized with dengue type 2 virus or vaccinia virus expressing the dengue type 4 virus NS3 protein were cross-reactive with dengue type 2 or type 4 virus, while broadly serotype cross-reactive CTL were a minority population. In dengue type 3 virus-immunized mice, the predominant CTL response to this epitope was broadly serotype cross reactive. All of the dengue virus-specific CTL clones studied also recognized the homologous NS3 sequences of one or more closely related flaviviruses, such as Kunjin virus. The critical contact residues for the CTL clones with different specificities were mapped with peptides having single amino acid substitutions. These data demonstrate that primary dengue virus infection induces a complex population of flavivirus-cross-reactive NS3-specific CTL clones in mice and suggest that CTL responses are influenced by the viral serotype. These findings suggest an additional mechanism by which the order of sequential flavivirus infections may influence disease manifestations. PMID- 9847342 TI - Intracellular traffic of herpes simplex virus glycoprotein gE: characterization of the sorting signals required for its trans-Golgi network localization. AB - Herpes simplex virus (HSV) and varicella-zoster virus (VZV) are two pathogenic human alphaherpesviruses whose intracellular assembly is thought to follow different pathways. VZV presumably acquires its envelope in the trans-Golgi network (TGN), and it has recently been shown that its major envelope glycoprotein, VZV-gE, accumulates in this compartment when expressed alone. In contrast, the envelopment of HSV has been proposed to occur at the inner nuclear membrane, although to which compartment the gE homolog (HSV-gE) is transported is unknown. For this reason, we have studied the intracellular traffic of HSV-gE and have found that this glycoprotein accumulates at steady state in the TGN, both when expressed from cloned cDNA and in HSV-infected cells. In addition, HSV-gE cycles between the TGN and the cell surface and requires a conserved tyrosine containing motif within its cytoplasmic tail for proper trafficking. These results show that VZV-gE and HSV-gE have similar intracellular trafficking pathways, probably reflecting the presence of similar sorting signals in the cytoplasmic domains of both molecules, and suggest that the respective viruses, VZV and HSV, could use the same subcellular organelle, the TGN, for their envelopment. PMID- 9847345 TI - Human cytomegalovirus inhibits transcription of the CC chemokine MCP-1 gene. AB - In primary human diploid fibroblasts, infection with an unpurified stock of human cytomegalovirus induced accumulation of the CC chemokine MCP-1 in the cell culture medium. By 24 h postinfection, the level of MCP-1 returned to that in uninfected cultures. When cells were infected with UV-inactivated human cytomegalovirus, the induction of MCP-1 was still observed, but no reduction was seen by 24 h postinfection or later. This effect was the result of a decrease in the level of MCP-1 mRNA present within the infected cell. Infection with purified virus revealed that the induction of MCP-1 was due to an activity found in the medium of infected cells; purified virions did not induce the expression of MCP 1. However, infection with purified virions repressed the level of MCP-1 mRNA below that found in uninfected cells. Additionally, infection with human cytomegalovirus prevented the induction of MCP-1 expression by tumor necrosis factor alpha and interleukin-1beta. The CC chemokine receptor encoded by the human cytomegalovirus US28 open reading frame (ORF) did not appear to play a role in this process, since a mutant virus in which the US28 ORF had been deleted downregulated MCP-1 in the same manner. PMID- 9847346 TI - Multiple nucleocapsid packaging of Autographa californica nucleopolyhedrovirus accelerates the onset of systemic infection in Trichoplusia ni. AB - Among the nucleopolyhedroviruses (Baculoviridae), the occlusion-derived virus (ODV), which initiates infection in host insects, may contain only a single nucleocapsid per virion (the SNPVs) or one to many nucleocapsids per virion (the MNPVs), but the significance of this difference is unclear. To gain insight into the biological relevance of these different packaging strategies, we compared pathogenesis induced by ODV fractions enriched for multiple nucleocapsids (ODV-M) or single nucleocapsids (ODV-S) of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) containing a beta-galactosidase reporter gene. In time course experiments wherein newly molted fourth-instar Trichoplusia ni were challenged with doses of ODV-S or ODV-M that yielded the same final mortality ( approximately 70%), we characterized viral foci as either being restricted to the midgut or involving tracheal cells (the secondary target tissue, indicative of systemic infection). We found that while the timing of primary infection by ODV-S and ODV-M was similar, ODV-S established significantly more primary midgut cell foci than ODV-M, but ODV-M infected tracheal cells at twice the rate of ODV-S. The more efficient establishment of tracheal infections by ODV-M decreased the probability that infections were lost by midgut cell sloughing, explaining why higher numbers of primary infections established by ODV-S within larvae were needed to achieve the same final mortality. These results showed that the multiple nucleocapsid packaging strategy of AcMNPV accelerates the onset of irreversible systemic infections and may indicate why MNPVs have wider individual host ranges than SNPVs. PMID- 9847347 TI - The ability of herpes simplex virus type 1 immediate-early protein Vmw110 to bind to a ubiquitin-specific protease contributes to its roles in the activation of gene expression and stimulation of virus replication. AB - Herpes simplex virus type 1 immediate-early protein Vmw110 stimulates the onset of virus infection and is required for efficient reactivation from latency. In transfection assays, Vmw110 is a potent activator of gene expression, but its mode of action has yet to be determined. Previous work has shown that Vmw110 localizes to specific intranuclear structures known as ND10, PML bodies, or PODs and causes the disruption of these domains. The ability of Vmw110 to disrupt ND10 correlates with its biological activities in infected and transfected cells. It has also been found that Vmw110 binds strongly and specifically to a ubiquitin specific protease known as HAUSP, itself a component of a subset of ND10. In this study we have investigated the role of HAUSP in Vmw110 activity; single amino acid residues of Vmw110 required for the interaction were identified, and the effects of mutation of these residues in infected and transfected cells were then assayed. The results indicate that the ability to bind to HAUSP contributes to the functional activities of Vmw110. PMID- 9847348 TI - Functional coupling between replication and packaging of poliovirus replicon RNA. AB - Poliovirus RNA genomes that contained deletions in the capsid-coding regions were synthesized in monkey kidney cells that constitutively expressed T7 RNA polymerase. These replication-competent subgenomic RNAs, or replicons (G. Kaplan and V. R. Racaniello, J. Virol. 62:1687-1696, 1988), were encapsidated in trans by superinfecting polioviruses. When superinfecting poliovirus resistant to the antiviral compound guanidine was used, the RNA replication of the replicon RNAs could be inhibited independently of the RNA replication of the guanidine resistant helper virus. Inhibiting the replication of the replicon RNA also profoundly inhibited its trans-encapsidation, even though the capsid proteins present in the cells could efficiently encapsidate the helper virus. The observed coupling between RNA replication and RNA packaging could account for the specificity of poliovirus RNA packaging in infected cells and the observed effects of mutations in the coding regions of nonstructural proteins on virion morphogenesis. It is suggested that this coupling results from direct interactions between the RNA replication machinery and the capsid proteins. The coupling of RNA packaging to RNA replication and of RNA replication to translation (J. E. Novak and K. Kirkegaard, Genes Dev. 8:1726-1737, 1994) could serve as mechanisms for late proofreading of poliovirus RNA, allowing only those RNA genomes capable of translating a full complement of functional RNA replication proteins to be propagated. PMID- 9847349 TI - Strong human immunodeficiency virus (HIV)-specific cytotoxic T-lymphocyte activity in Sydney Blood Bank Cohort patients infected with nef-defective HIV type 1. AB - Proposals for the use of live attenuated human immunodeficiency virus (HIV) type 1 (HIV-1) as a vaccine candidate in humans have been based on the protection afforded by attenuated simian immunodeficiency virus in the macaque model. Although it is not yet known if this strategy could succeed in humans, a study of the Sydney Blood Bank Cohort (SBBC), infected with an attenuated HIV-1 quasispecies with natural nef and nef/long terminal repeat deletions for up to 17 years, could provide insights into the long-term immunological consequences of living with an attenuated HIV-1 infection. In this study, HIV-specific cytoxic T lymphocyte (CTL) responses in an SBBC donor and six recipients were examined over a 3-year period with enzyme-linked immunospot, tetrameric complex binding, direct CTL lysis, and CTL precursor level techniques. Strong HIV-specific CTL responses were detected in four of seven patients, including one patient with an undetectable viral load. Two of seven patients had weak CTL responses, and in one recipient, no HIV-specific CTLs were detected. High levels of circulating effector and memory HIV-specific CTLs can be maintained for prolonged periods in these patients despite very low viral loads. PMID- 9847350 TI - Overlapping signals for transcription and replication at the 3' terminus of the vesicular stomatitis virus genome. AB - Transcription and replication signals within the negative-sense genomic RNA of vesicular stomatitis virus (VSV) are located at the 3' terminus. To identify these signals, we have used a transcription- and replication-competent minigenome of VSV to generate a series of deletions spanning the first 47 nucleotides at the 3' terminus of the VSV genome corresponding to the leader gene. Analysis of these mutants for their ability to replicate showed that deletion of sequences within the first 24 nucleotides abrogated or greatly reduced the level of replication. Deletion of downstream sequences from nucleotides 25 to 47 reduced the level of replication only to 55 to 70% of that of the parental template. When transcription activity of these templates was measured, the first 24 nucleotides were also found to be required for transcription, since deletion of these sequences blocked or significantly reduced transcription. Downstream sequences from nucleotides 25 to 47 were necessary for optimal levels of transcription. Furthermore, replacement of sequences within the 25 to 47 nucleotides with random heterologous nonviral sequences generated mutant templates that replicated well (65 to 70% of the wild-type levels) but were transcribed poorly (10 to 15% of the wild-type levels). These results suggest that the minimal promoter for transcription and replication could be as small as the first 19 nucleotides and is contained within the 3'-terminal 24 nucleotides of the VSV genome. The sequences from nucleotides 25 to 47 may play a more important role in optimal transcription than in replication. Our results also show that deletion of sequences within the leader gene does not influence the site of transcription reinitiation of the downstream gene. PMID- 9847352 TI - Altered growth characteristics of recombinant respiratory syncytial viruses which do not produce NS2 protein. AB - The second gene in the 3'-to-5' gene order in respiratory syncytial virus (RSV) encodes the nonstructural protein NS2, for which there is no assigned function. To study the function of NS2, we have used a recently developed reverse genetics system to ablate expression of NS2 in recombinant RSV. A full-length cDNA copy of the antigenome of RSV A2 strain under the control of a T7 promoter was modified by introduction of tandem termination codons within the NS2 open reading frame (NS2stop) or by deletion of the entire NS2 gene (DeltaNS2). The NS2 knockout antigenomic cDNAs were cotransfected with plasmids encoding the N, P, L, and M2-1 proteins of RSV, each controlled by the T7 promoter, into cells infected with a vaccinia virus recombinant expressing T7 RNA polymerase. Recombinant NS2stop and DeltaNS2 RSVs were recovered and characterized. Both types of NS2 knockout virus displayed pinpoint plaque morphology and grew more slowly than wild-type RSV. The expression of monocistronic mRNAs for the five genes examined (NS1, NS2, N, F, and L) was unchanged in cells infected with either type of NS2 knockout virus, except that no NS2 mRNA was detected with the DeltaNS2 virus. Synthesis of readthrough mRNAs was affected only for the DeltaNS2 virus, where the NS1-NS2, NS2-N, and NS1-NS2-N mRNAs were replaced with the predicted novel NS1-N mRNA. Upon passage, the NS2stop virus stock rapidly developed revertants which expressed NS2 protein and grew with similar plaque morphology and kinetics wild type RSV. Sequence analysis confirmed that the termination codons had reverted to sense, albeit not the wild-type assignments, and provided evidence consistent with biased hypermutation. No revertants were recovered from recombinant DeltaNS2 RSV. These results show that the NS2 protein is not essential for RSV replication, although its presence greatly improves virus growth in cell culture. The attenuated phenotype of these mutant viruses, coupled with the expected genetic stability associated with gene deletions, suggests that the DeltaNS2 RSV is a candidate for vaccine development. PMID- 9847351 TI - Mutations in nonconserved domains of Ty3 integrase affect multiple stages of the Ty3 life cycle. AB - Ty3, a retroviruslike element of Saccharomyces cerevisiae, transposes into positions immediately upstream of RNA polymerase III-transcribed genes. The Ty3 integrase (IN) protein is required for integration of the replicated, extrachromosomal Ty3 DNA. In retroviral IN, a conserved core region is sufficient for strand transfer activity. In this study, charged-to-alanine scanning mutagenesis was used to investigate the roles of the nonconserved amino- and carboxyl-terminal regions of Ty3 IN. Each of the 20 IN mutants was defective for transposition, but no mutant was grossly defective for capsid maturation. All mutations affecting steady-state levels of mature IN protein resulted in reduced levels of replicated DNA, even when polymerase activity was not grossly defective as measured by exogenous reverse transcriptase activity assay. Thus, IN could contribute to nonpolymerase functions required for DNA production in vivo or to the stability of the DNA product. Several mutations in the carboxyl-terminal domain resulted in relatively low levels of processed 3' ends of the replicated DNA, suggesting that this domain may be important for binding of IN to the long terminal repeat. Another class of mutants produced wild-type amounts of DNA with correctly processed 3' ends. This class could include mutants affected in nuclear entry and target association. Collectively, these mutations demonstrate that in vivo, within the preintegration complex, IN performs a central role in coordinating multiple late stages of the retrotransposition life cycle. PMID- 9847354 TI - Focal transcriptional activity of murine cytomegalovirus during latency in the lungs. AB - Interstitial pneumonia is a frequent and critical manifestation of human cytomegalovirus (CMV) disease in immunocompromised patients, in particular in recipients of bone marrow transplantation. Previous work in the murine CMV infection model has identified the lungs as a major organ site of CMV latency and recurrence. It was open to question whether the viral genome is transcriptionally silent or active during latency. Transcription could be latency associated and thus be part of the latency phenotype. Alternatively, transcriptional activity could reflect episodes of reactivation. We demonstrate here that transcription of the immediate-early (IE) transcription unit ie1-ie3 selectively generates ie1 specific transcripts during latency. Notably, while the latent viral DNA was found to be evenly distributed in the lungs, transcription was focal and randomly distributed. This finding indicates that IE transcription is not a feature inherent to murine CMV latency but rather reflects foci of primordial reactivation. However, this reactivation did not initiate productive infection, since ie3 gene mRNA specifying the essential transactivator IE3 of murine CMV early gene expression was not detectable. Accordingly, transcripts encoding gB were absent during latency. By contrast, during induced virus recurrence, IE phase transcription switched from focal to generalized and ie3-specific transcripts were generated. These data imply that latency and recurrence are regulated not only at the IE promoter-enhancer and that there exists an additional checkpoint at the level of precursor RNA splicing. We propose that focal transcription reflects random episodes of nonproductive reactivation that get terminated before IE3 is expressed and ignites the productive cycle. PMID- 9847353 TI - Hepatitis B virus RNA-binding proteins associated with cytokine-induced clearance of viral RNA from the liver of transgenic mice. AB - Hepatitis B virus (HBV) gene expression is downregulated in the liver of HBV transgenic mice by a posttranscriptional mechanism that is triggered by the local production of gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF alpha) during intrahepatic inflammation (hepatitis). The molecular basis for this antiviral effect is unknown. In this study, we identified three HBV RNA-binding liver nuclear proteins (p45, p39, and p26) the relative abundance of which correlates with the abundance of HBV RNA in response to the induction of IFN gamma and TNF-alpha. All three proteins bind to a 91-bp element located at the 5' end of a previously defined posttranscriptional regulatory element that is thought to mediate the nuclear export of HBV RNA. The presence of p45 correlates directly with the presence of HBV RNA, being detectable under baseline conditions when the viral RNA is abundant and undetectable when the viral RNA disappears in response to IFN-gamma and TNF-alpha. In contrast, p26 is inversely related to HBV RNA, being detectable only when the viral RNA disappears following cytokine activation. Finally, p39 is constitutively expressed, and its abundance and mobility appear to be slightly increased by cytokine activation. These results suggest a model in which hepatocellular HBV RNA content might be controlled by the stabilizing and/or destabilizing influences of these RNA-binding proteins whose activity is regulated by cytokine-induced signaling pathways. PMID- 9847355 TI - Efficient transduction by an amphotropic retrovirus vector is dependent on high level expression of the cell surface virus receptor. AB - Transduction by murine leukemia virus-based retrovirus vectors is limited in certain cell types, particularly in nondividing cells. But transduction can be inefficient even in cells that divide rapidly. For example, exposure of 208F rat embryo fibroblasts to an excess of an amphotropic retrovirus vector encoding alkaline phosphatase results in a transduction efficiency of only about 10%, even though these cells divide rapidly. Here we show that transduction of 208F cells is limited by cell surface retrovirus receptor levels; overexpression of the amphotropic retrovirus receptor Pit2 markedly improved the transduction efficiency to 50%. To characterize receptor levels and binding affinity, we synthesized a fusion protein that joins the amino terminus of the amphotropic envelope protein to the Fc region of a human immunoglobulin G1 molecule for use in binding assays. In comparison to the parental cell line, the modified cell line showed an order of magnitude increase in binding sites of from 18,000 to 150,000 per cell. Thus, efficient transduction by an amphotropic retrovirus vector requires high-level expression of the retrovirus receptor Pit2. These results provide the rationale for further examination of the role of receptor levels in inefficient transduction, especially with regard to target cells for gene therapy, where a high transduction rate is often crucial. PMID- 9847356 TI - In vivo modulation of vaccine-induced immune responses toward a Th1 phenotype increases potency and vaccine effectiveness in a herpes simplex virus type 2 mouse model. AB - Several vaccines have been investigated experimentally in the herpes simplex virus type 2 (HSV-2) model system. While it is believed that CD4(+)-T-cell responses are important for protection in general, the correlates of protection from HSV-2 infection are still under investigation. Recently, the use of molecular adjuvants to drive vaccine responses induced by DNA vaccines has been reported in a number of experimental systems. We sought to take advantage of this immunization model to gain insight into the correlates of immune protection in the HSV-2 mouse model system and to further explore DNA vaccine technology. To investigate whether the Th1- or Th2-type immune responses are more important for protection from HSV-2 infection, we codelivered the DNA expression construct encoding the HSV-2 gD protein with the gene plasmids encoding the Th1-type (interleukin-2 [IL-2], IL-12, IL-15, and IL-18) and Th2-type (IL-4 and IL-10) cytokines in an effort to drive immunity induced by vaccination. We then analyzed the modulatory effects of the vaccine on the resulting immune phenotype and on the mortality and the morbidity of the immunized animals following a lethal challenge with HSV-2. We observed that Th1 cytokine gene coadministration not only enhanced the survival rate but also reduced the frequency and severity of herpetic lesions following intravaginal HSV challenge. On the other hand, coinjection with Th2 cytokine genes increased the rate of mortality and morbidity of the challenged mice. Moreover, of the Th1-type cytokine genes tested, IL-12 was a particularly potent adjuvant for the gD DNA vaccination. PMID- 9847357 TI - Pathogenicity of different rabies virus variants inversely correlates with apoptosis and rabies virus glycoprotein expression in infected primary neuron cultures. AB - The mouse-adapted rabies virus strain CVS-24 has stable variants, CVS-B2c and CVS N2c, which differ greatly in their pathogenicity for normal adult mice and in their ability to infect nonneuronal cells. The glycoprotein (G protein), which has previously been implicated in rabies virus pathogenicity, shows substantial structural differences between these variants. Although prior studies have identified antigenic site III of the G protein as the major pathogenicity determinant, CVS-B2c and CVS-N2c do not vary at this site. The possibility that pathogenicity is inversely related to G protein expression levels is suggested by the finding that CVS-B2c, the less pathogenic variant, expresses at least fourfold-higher levels of G protein than CVS-N2c in infected neurons. Although there is some difference between CVS-B2c- and CVS-N2c-infected neurons in G protein mRNA expression levels, the differential expression of G protein appears to be largely determined by posttranslational mechanisms that affect G protein stability. Pulse-chase experiments indicated that the G protein of CVS-B2c is degraded more slowly than that of CVS-N2c. The accumulation of G protein correlated with the induction of programmed cell death in CVS-B2c-infected neurons. The extent of apoptosis was considerably lower in CVS-N2c-infected neurons, where G protein expression was minimal. While nucleoprotein (N protein) expression levels were similar in neurons infected with either variant, the transport of N protein into neuronal processes was strongly inhibited in CVS-B2c infected cells. Thus, downregulation of G protein expression in neuronal cells evidently contributes to rabies virus pathogenesis by preventing apoptosis and the apparently associated failure of the axonal transport of N protein. PMID- 9847358 TI - Herpes simplex virus type 1 vector-mediated expression of nerve growth factor protects dorsal root ganglion neurons from peroxide toxicity. AB - Nerve growth factor beta subunit (beta-NGF) transgene delivery and expression by herpes simplex virus type 1 (HSV-1) vectors was examined in a cell culture model of neuroprotection from hydrogen peroxide toxicity. Replication-competent (tk- K mutant background) and replication-defective (ICP4(-);tk- S mutant background) vectors were engineered to contain the murine beta-NGF cDNA under transcriptional control of either the human cytomegalovirus immediate-early gene promoter (HCMV IEp) (e.g., KHN and SHN) or the latency-active promoter 2 (LAP2) (e.g., KLN and SLN) within the viral thymidine kinase (tk) locus. Infection of rat B103 and mouse N2A neuronal cell lines, 9L rat glioma cells, and Vero cells with the KHN or SHN vectors resulted in the production of beta-NGF-specific transcripts and beta-NGF protein reaching a maximum at 3 days postinfection (p.i.). NGF protein was released into the culture media in amounts ranging from 10.83 to 352.86 ng/ml, with the highest levels being achieved in B103 cells, and was capable of inducing neurite sprouting of PC-12 cells. The same vectors produced high levels of NGF in primary dorsal root ganglion (DRG) cultures at 3 days. In contrast to HCMV IEp-mediated expression, the LAP2-NGF vectors showed robust expression in primary DRG neurons at 14 days. The neuroprotective effect of vector produced NGF was assessed by its ability to inhibit hydrogen peroxide-induced neuron toxicity in primary DRG cultures. Consistent with the kinetics of vector-mediated NGF expression, HCMV-NGF vectors were effective in abrogating the toxic effects of peroxide at 3 but not 14 days p.i. whereas LAP2-NGF vector transduction inhibited apoptosis in DRG neurons at 14 days p.i. but was ineffective at 3 days p.i. Similar kinetics of NGF expression were observed with the KHN and KLN vectors in latently infected mouse trigeminal ganglia, where high levels of beta-NGF protein expression were detected at 4 wks p.i. only from the LAP2; HCMV-NGF-driven expression peaked at 3 days but could not be detected during HSV latency at 4 weeks. Together, these results indicate that (i) NGF vector-infected cells produce and secrete mature, biologically active beta-NGF; (ii) vector-synthesized NGF was capable of blocking peroxide-induced apoptosis in primary DRG cultures; and (iii) the HCMV-IEp functioned to produce high levels of NGF for several days; but (iv) only the native LAP2 was capable of long-term expression of a therapeutic gene product in latently infected neurons in vivo. PMID- 9847360 TI - Nuclear targeting of the cauliflower mosaic virus coat protein. AB - The entry of the viral genomic DNA of cauliflower mosaic virus into the nucleus is a critical step of viral infection. We have shown by transient expression in plant protoplasts that the viral coat protein (CP), which is processed from the product of open reading frame IV, contains an N-terminal nuclear localization signal (NLS). The NLS is exposed on the surface of the virion and is thus available for interaction with a putative NLS receptor. Phosphorylation of the matured CP did not influence the nuclear localization of the protein but improved protein stability. Mutation of the NLS completely abolished viral infectivity, thus indicating its importance in the virus life cycle. The NLS seems to be regulated by the N terminus of the precapsid, which inhibits its nuclear targeting. This regulation could be important in allowing virus assembly in the cytoplasm. PMID- 9847361 TI - Selection of functional variants of the NS3-NS4A protease of hepatitis C virus by using chimeric sindbis viruses. AB - The NS3-NS4A serine protease of hepatitis C virus (HCV) mediates four specific cleavages of the viral polyprotein and its activity is considered essential for the biogenesis of the HCV replication machinery. Despite extensive biochemical and structural characterization, the analysis of natural variants of this enzyme has been limited by the lack of an efficient replication system for HCV in cultured cells. We have recently described the generation of chimeric HCV-Sindbis viruses whose propagation depends on the NS3-NS4A catalytic activity. NS3-NS4A gene sequences were fused to the gene coding for the Sindbis virus structural polyprotein in such a way that processing of the chimeric polyprotein, nucleocapsid assembly, and production of infectious viruses required NS3-NS4A mediated proteolysis (G. Filocamo, L. Pacini, and G. Migliaccio, J. Virol. 71:1417-1427, 1997). Here we report the use of these chimeric viruses to select and characterize active variants of the NS3-NS4A protease. Our original chimeric viruses displayed a temperature-sensitive phenotype and formed lysis plaques much smaller than those formed by wild-type (wt) Sindbis virus. By serially passaging these chimeric viruses on BHK cells, we have selected virus variants which formed lysis plaques larger than those produced by their progenitors and produced NS3 NS4A proteins different in size and/or sequence from those of the original viruses. Characterization of the selected protease variants revealed that all of the mutated proteases still efficiently processed the chimeric polyprotein in infected cells and also cleaved an HCV substrate in vitro. One of the selected proteases was expressed in a bacterial system and showed a catalytic efficiency comparable to that of the wt recombinant protease. PMID- 9847362 TI - A packaging cell line for lentivirus vectors. AB - Lentivirus vectors can transduce dividing and nondividing cells. Using three plasmid transient transfections, high-titer (>10(9) IU/ml) recombinant lentivirus vectors pseudotyped with vesicular stomatitis virus G (VSV-G) protein can be generated (T. Kafri et al., Nat. Genet. 17:314-317, 1997; H. Miyoshi et al., Proc. Natl. Acad. Sci. USA 94:10319-10323, 1997; L. Naldini et al., Science 272:263-267, 1996). The recombinant lentiviruses can efficiently infect brain, liver, muscle, and retinal tissue in vivo. Furthermore, the transduced tissues demonstrated long-term expression of reporter genes in immunocompetent rodents. We now report the generation of a tetracycline-inducible VSV-G pseudotyped lentivirus packaging cell line which can generate virus particles at titers greater than 10(6) IU/ml for at least 3 to 4 days. The vector produced by the inducible cell line can be concentrated to titers of 10(9) IU/ml and can efficiently transduce nondividing cells in vitro and in vivo. The availability of a lentivirus packaging cell line will significantly facilitate the production of high-titer lentivirus vectors for gene therapy and study of human immunodeficiency virus biology. PMID- 9847359 TI - The genome of Melanoplus sanguinipes entomopoxvirus. AB - The family Poxviridae contains two subfamilies: the Entomopoxvirinae (poxviruses of insects) and the Chordopoxvirinae (poxviruses of vertebrates). Here we present the first characterization of the genome of an entomopoxvirus (EPV) which infects the North American migratory grasshopper Melanoplus sanguinipes and other important orthopteran pests. The 236-kbp M. sanguinipes EPV (MsEPV) genome consists of a central coding region bounded by 7-kbp inverted terminal repeats and contains 267 open reading frames (ORFs), of which 107 exhibit similarity to previously described genes. The presence of genes not previously described in poxviruses, and in some cases in any other known virus, suggests significant viral adaptation to the arthropod host and the external environment. Genes predicting interactions with host cellular mechanisms include homologues of the inhibitor of apoptosis protein, stress response protein phosphatase 2C, extracellular matrixin metalloproteases, ubiquitin, calcium binding EF-hand protein, glycosyltransferase, and a triacylglyceride lipase. MsEPV genes with putative functions in prevention and repair of DNA damage include a complete base excision repair pathway (uracil DNA glycosylase, AP endonuclease, DNA polymerase beta, and an NAD+-dependent DNA ligase), a photoreactivation repair pathway (cyclobutane pyrimidine dimer photolyase), a LINE-type reverse transcriptase, and a mutT homologue. The presence of these specific repair pathways may represent viral adaptation for repair of environmentally induced DNA damage. The absence of previously described poxvirus enzymes involved in nucleotide metabolism and the presence of a novel thymidylate synthase homologue suggest that MsEPV is heavily reliant on host cell nucleotide pools and the de novo nucleotide biosynthesis pathway. MsEPV and lepidopteran genus B EPVs lack genome colinearity and exhibit a low level of amino acid identity among homologous genes (20 to 59%), perhaps reflecting a significant evolutionary distance between lepidopteran and orthopteran viruses. Divergence between MsEPV and the Chordopoxvirinae is indicated by the presence of only 49 identifiable chordopoxvirus homologues, low level amino acid identity among these genes (20 to 48%), and the presence in MsEPV of 43 novel ORFs in five gene families. Genes common to both poxvirus subfamilies, which include those encoding enzymes involved in RNA transcription and modification, DNA replication, protein processing, virion assembly, and virion structural proteins, define the genetic core of the Poxviridae. PMID- 9847363 TI - Sin Nombre virus pathogenesis in Peromyscus maniculatus. AB - Sin Nombre virus (SNV), a member of the Hantavirus genus, causes acute viral pneumonia in humans and is thought to persistently infect mice. The deer mouse, Peromyscus maniculatus, has been identified as the primary reservoir host for SNV. To understand SNV infection of P. maniculatus, we examined wild deer mice for localization of viral antigens and nucleic acid. Morphologic examination consistently revealed septal edema within lung tissue and mononuclear cell infiltrates in portal areas of the liver. Immunohistochemical analysis of SNV infected deer mice identified viral antigens within lung, liver, kidney, and spleen. The lungs consistently presented with the highest levels of viral antigen by immunohistochemistry and with the highest levels of nucleic acid by reverse transcriptase (RT) PCR. The mononuclear cell infiltrates surrounding liver portal triads were positive for SNV antigens in addition to resident macrophages in liver sinuses. Spleen tissue contained antigens in both the red pulp and the periartereolar region of the white pulp. The kidney presented with no gross pathology, although antigens could be localized to glomeruli. Virus antigen levels within the kidney were highest in deer mice that did not have antibodies to SNV but contained viral nucleic acid detectable by RT PCR. Since transmission is thought to occur via urine, our results suggest that virus transmission may be highest in the early stages of infection. In addition, these results indicate that SNV does cause some pathology within its reservoir host. PMID- 9847364 TI - Interaction with the p6 domain of the gag precursor mediates incorporation into virions of Vpr and Vpx proteins from primate lentiviruses. AB - Vpr and Vpx proteins from human and simian immunodeficiency viruses (HIV and SIV) are incorporated into virions in quantities equivalent to those of the viral Gag proteins. We demonstrate here that Vpr and Vpx proteins from distinct lineages of primate lentiviruses were able to bind to their respective Gag precursors. The capacity of HIV type 1 (HIV-1) Vpr mutants to bind to Pr55(Gag) was correlated with their incorporation into virions. Molecular analysis of these interactions revealed that they required the C-terminal p6 domain of the Gag precursors. While the signal for HIV-1 Vpr binding lies in the leucine triplet repeat region of the p6 domain reported to be essential for incorporation, SIVsm Gag lacking the equivalent region still bound to SIVsm Vpr and Vpx, indicating that the determinants for Gag binding are located upstream of this region of the p6 domain. Binding to Gag cleavage products showed that HIV-1 Vpr interacted directly with the nucleocapsid protein (NC), whereas SIVsm Vpr and Vpx did not interact with NC but with the p6 protein. These results (i) reveal differences between HIV-1 and SIVsm for the p6 determinants required for Vpr and Vpx binding to Gag and (ii) suggest that HIV-1 Vpr and SIVsm Vpr and Vpx interact with distinct cleavage products of the precursor following proteolytic processing in the virions. PMID- 9847365 TI - Asymptomatic simian immunodeficiency virus infection decreases blood CD4(+) T cells by accumulating recirculating lymphocytes in the lymphoid tissues. AB - Declining blood CD4(+) T-cell counts mark the progress of simian immunodeficiency virus (SIV) disease in macaques and model the consequences of untreated human immunodeficiency virus infection in humans. However, blood lymphocytes are only a fraction of the recirculating lymphocyte pool, and their numbers are affected by cell synthesis, cell depletion, and distribution among blood and lymphoid tissue compartments. Asymptomatic, SIV-infected macaques maintained constant and nearly normal numbers of recirculating lymphocytes despite the decline in CD4(+) T-cell counts. Substantial depletion was detected only when blood CD4(+) T-cell counts fell below 300/microliter. In asymptomatic animals, changes in CD4(+) T-cell distribution were more important than lymphocyte depletion for controlling the blood cell levels. PMID- 9847366 TI - Demonstration that orf2 encodes the feline immunodeficiency virus transactivating (Tat) protein and characterization of a unique gene product with partial rev activity. AB - The long PCR technique was used to amplify the three size classes of viral mRNAs produced in cells infected by feline immunodeficiency virus (FIV). We identified in the env region a new splice acceptor site that generated two transcripts, each coding for an 11-kDa protein, p11(rev), whose function is unknown. The small-size class of mRNAs included two bicistronic orf2/rev mRNAs and two rev-like mRNAs, consisting only of the second exon of rev and coding for a 15-kDa protein, p15(rev). p15(rev) contained the minimal effector domain of Rev and was sufficient to mediate partial Rev activity. The bicistronic mRNAs encoded two distinct proteins, one of 23 kDa corresponding to Rev and a 9-kDa protein encoded by the orf2 gene. The orf2 gene product is a protein of 79 amino acids with characteristics similar to those of the Tat (transactivator) proteins of the ungulate lentiviruses. Transient expression assays, using the FIV long terminal repeat (LTR) to drive transcription of the bacterial gene for chloramphenicol acetyltransferase demonstrated that the orf2 gene transactivates gene expression an average of 14- to 20-fold above the basal level. Deletion mutants of the FIV LTR were generated to locate sequences responsive to transactivation mediated by the orf2 gene. A 5' deletion mutant that removed the AP1 site resulted in residual low-level transactivation by orf2. Further experiments using LTR mutants with internal deletions identified three regions located between positions -126 and -47 relative to the cap site that were important for orf2-directed transactivation. These regions include the AP1 site, a C/EBP tandem repeat, and an ATF site. PMID- 9847367 TI - Limited breadth of the protective immunity elicited by simian immunodeficiency virus SIVmne gp160 vaccines in a combination immunization regimen. AB - We previously reported that immunization with recombinant simian immunodeficiency virus SIVmne envelope (gp160) vaccines protected macaques against an intravenous challenge by the cloned homologous virus, E11S. In this study, we confirmed this observation and found that the vaccines were effective not only against virus grown on human T-cell lines but also against virus grown on macaque peripheral blood mononuclear cells (PBMC). The breadth of protection, however, was limited. In three experiments, 3 of 10 animals challenged with the parental uncloned SIVmne were completely protected. Of the remaining animals, three were transiently virus positive and four were persistently positive after challenge, as were 10 nonimmunized control animals. Protection was not correlated with levels of serum-neutralizing antibodies against the homologous SIVmne or a related virus, SIVmac251. To gain further insight into the protective mechanism, we analyzed nucleotide sequences in the envelope region of the uncloned challenge virus and compared them with those present in the PBMC of infected animals. The majority (85%) of the uncloned challenge virus was homologous to the molecular clone from which the vaccines were made (E11S type). The remaining 15% contained conserved changes in the V1 region (variant types). Control animals infected with this uncloned virus had different proportions of the two genotypes, whereas three of four immunized but persistently infected animals had >99% of the variant types early after infection. These results indicate that the protective immunity elicited by recombinant gp160 vaccines is restricted primarily to the homologous virus and suggest the possibility that immune responses directed to the V1 region of the envelope protein play a role in protection. PMID- 9847368 TI - Characterization of influenza virus PB1 protein binding to viral RNA: two separate regions of the protein contribute to the interaction domain. AB - The interaction of the PB1 subunit of the influenza virus polymerase with the viral RNA (vRNA) template has been studied in vitro. The experimental approach included the in vitro binding of labeled model vRNA to PB1 protein immobilized as an immunoprecipitate, as well as Northwestern analyses. The binding to model vRNA was specific, and an apparent Kd of about 2 x 10(-8) M was determined. Although interaction with the isolated 3' arm of the panhandle was detectable, interaction with the 5' arm was prominent and the binding was optimal with a panhandle analog structure (5'+3' probe). When presented with a panhandle analog mixed probe, PB1 was able to retain the 3' arm as efficiently as the 5' arm. The sequences of the PB1 protein involved in vRNA binding were identified by in vitro interaction tests with PB1 deletion mutants. Two separate regions of the PB1 protein sequence proved positive for binding: the N-terminal 83 amino acids and the C-proximal sequences located downstream of position 493. All mutants able to interact with model vRNA were capable of binding the 5' arm more efficiently than the 3' arm of the panhandle. Taken together, these results suggest that two separate regions of the PB1 protein constitute a vRNA binding site that interacts preferentially with the 5' arm of the panhandle structure. PMID- 9847369 TI - Persistent infection promotes cross-species transmissibility of mouse hepatitis virus. AB - Persistent infection with mouse hepatitis virus (MHV) strain A59 in murine DBT (delayed brain tumor) cells resulted in the emergence of host range variants, designated V51A and V51B, at 210 days postinfection. These host range mutants replicated efficiently in normally nonpermissive Chinese hamster ovary (CHO), in human hepatocarcinoma (HepG2), and to a lesser extent in human breast carcinoma (MCF7) cell lines. Little if any replication was noted in baby hamster kidney (BHK), green African monkey kidney (COS-7), feline kidney (CRFK), and swine testicular (ST) cell lines. By fluorescent antibody (FA) staining, persistent viruses V10B and V30B, isolated at days 38 and 119 days postinfection, also demonstrated very low levels of replication in human HepG2 cells. These data suggest that persistence may rapidly select for host range expansion of animal viruses. Pretreatment of HepG2 cells with a polyclonal antibody directed against human carcinoembryonic antigens (CEA) or with some monoclonal antibodies (Col-1, Col-4, Col-12, and Col-14) that bind human CEA significantly inhibited V51B infection. Under identical conditions, little or no blockade was evident with other monoclonal antibodies (kat4c or Col-6) which also bind the human CEA glycoproteins. In addition, an antibody (EDDA) directed against irrelevant antigens did not block V51B replication. Pretreatment with the Col-4 and Col-14 antibodies did not block Sindbis virus replication in HepG2 cells or MHV infection in DBT cells, suggesting that one or more CEA glycoproteins likely functioned as receptors for V51B entry into human cell lines. To test this hypothesis, the human biliary glycoprotein (Bgp) and CEA genes were cloned and expressed in normally nonpermissive BHK cell lines by using noncytopathic Sindbis virus replicons (pSinRep19). By growth curves and FA staining, human CEA and to a much lesser extent human Bgp functioned as receptors for V51B entry. Furthermore, V51B replication was blocked with polyclonal antiserum directed against human CEA and Bgp. Under identical conditions, the parental MHV strain A59 failed to replicate in BHK cells expressing human Bgp or CEA. These data suggest that MHV persistence may promote virus cross-species transmissibility by selecting for virus variants that recognize phylogenetic homologues of the normal receptor. PMID- 9847370 TI - Herpes simplex virus type 1 immediate-early protein vmw110 induces the proteasome dependent degradation of the catalytic subunit of DNA-dependent protein kinase. AB - Herpes simplex virus type 1 (HSV-1) infection causes the active degradation of the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), and this process is reliant on the expression of the HSV-1 immediate-early protein Vmw110. In this study we investigated in more detail the mechanism by which the degradation occurs, the domains of Vmw110 which are required, and whether Vmw110 is by itself sufficient for the effect. We found that proteasome inhibitors prevented the degradation of DNA-PKcs, indicating the involvement of a proteasome pathway. Furthermore, the continued activity of DNA-PK during infection in the presence of these inhibitors indicated that Vmw110 does not directly alter the enzyme activity of DNA-PKcs prior to its degradation in a normal infection. Indeed, Vmw110 was found to bind to neither the catalytic nor Ku subunits of DNA PK. Using mutant Vmw110 viruses we show that the RING finger domain of Vmw110 is essential for the induced degradation of DNA-PKcs but that the ability of Vmw110 to bind to a cellular ubiquitin-specific protease (HAUSP) is not required. When expressed in the absence of other viral proteins, Vmw110 was sufficient to cause the degradation of DNA-PKcs, indicating that the effect on the stability of DNA PKcs was a direct consequence of Vmw110 activity and not an indirect Vmw110 dependent effect of virus infection. Finally, the Vmw110-induced degradation of DNA-PKcs and loss in DNA-PK activity appears to be beneficial to HSV-1 infection, as virus replication was more efficient in cells lacking DNA-PKcs, especially at low multiplicities of infection. PMID- 9847371 TI - Characterization of the Jembrana disease virus tat gene and the cis- and trans regulatory elements in its long terminal repeats. AB - Jembrana disease virus (JDV) is a newly identified bovine lentivirus that is closely related to the bovine immunodeficiency virus (BIV). JDV contains a tat gene, encoded by two exons, which has potent transactivation activity. Cotransfection of the JDV tat expression plasmid with the JDV promoter chloramphenicol acetyltransferase (CAT) construct pJDV-U3R resulted in a substantial increase in the level of CAT mRNA transcribed from the JDV long terminal repeat (LTR) and a dramatic increase in the CAT protein level. Deletion analysis of the LTR sequences showed that sequences spanning nucleotides -68 to +53, including the TATA box and the predicted first stem-loop structure of the predicted Tat response element (TAR), were required for efficient transactivation. The results, derived from site-directed mutagenesis experiments, suggested that the base pairing in the stem of the first stem-loop structure in the TAR region was important for JDV Tat-mediated transactivation; in contrast, nucleotide substitutions in the loop region of JDV TAR had less effect. For the JDV LTR, upstream sequences, from nucleotide -196 and beyond, as well as the predicted secondary structures in the R region, may have a negative effect on basal JDV promoter activity. Deletion of these regions resulted in a four- to fivefold increase in basal expression. The JDV Tat is also a potent transactivator of other animal and primate lentivirus promoters. It transactivated BIV and human immunodeficiency virus type 1 (HIV-1) LTRs to levels similar to those with their homologous Tat proteins. In contrast, HIV-1 Tat has minimal effects on JDV LTR expression, whereas BIV Tat moderately transactivated the JDV LTR. Our study suggests that JDV may use a mechanism of transactivation similar but not identical to those of other animal and primate lentiviruses. PMID- 9847372 TI - Recombination in Tula hantavirus evolution: analysis of genetic lineages from Slovakia. AB - To examine the evolution of Tula hantavirus (TUL), carried by the European common vole (Microtus arvalis and M. rossiaemeridionalis), we have analyzed genetic variants from Slovakia, the country where the virus is endemic. Phylogenetic analysis (PHYLIP) based on either partial (nucleotides [nt] 441 to 898) or complete N-protein-encoding sequences divided Slovakian TUL variants into two main lineages: (i) strains from eastern Slovakia, which clustered with Russian strains, and (ii) strains from western Slovakia situated closer to those from the Czech Republic. We found genetic diversity of 19% between the two groups and 4% within the western Slovakian TUL strains. Phylogenetic analysis of the 3' noncoding region (3'-NCR), however, placed the eastern Slovakian strains closer to those from western Slovakia and the Czech Republic, with a greater distance to the Russian strains, suggesting a recombinant nature of the S segment in the eastern Slovakian TUL lineage. A bootscan search of the S-segment sequences of TUL strains revealed at least two recombination points in the S sequences of eastern Slovakian TUL strains (nt 400 to 415 and around 1200) which agreed well with the pattern of amino acid substitutions in the N protein and deletions/insertions in the 3'-NCR of the S segment. These data suggest that homologous recombination events occurred in the evolution of hantaviruses. PMID- 9847373 TI - Human cytomegalovirus UL69 protein induces cells to accumulate in G1 phase of the cell cycle. AB - Earlier studies have revealed that human cytomegalovirus rapidly inhibits the growth of fibroblasts, blocking cell cycle progression at multiple points, including the G1-to-S-phase transition. The present study demonstrates that the UL69 protein, a virus-encoded constituent of the virion, is able to arrest cell cycle progression when introduced into uninfected cells. Expression of the UL69 protein causes U2 OS cells and primary human fibroblasts to accumulate within the G1 compartment of the cell cycle, and serum fails to induce the progression of quiescent human fibroblasts into the S phase when the protein is present. Therefore, the UL69 protein is at least partially responsible for the cell cycle block that is instituted after infection of permissive cells with human cytomegalovirus. PMID- 9847374 TI - Enhancement of human immunodeficiency virus type 1 infection by the CC-chemokine RANTES is independent of the mechanism of virus-cell fusion. AB - We have studied the effects of CC-chemokines on human immunodeficiency virus type 1 (HIV-1) infection, focusing on the infectivity enhancement caused by RANTES. High RANTES concentrations increase the infectivity of HIV-1 isolates that use CXC-chemokine receptor 4 for entry. However, RANTES can have a similar enhancing effect on macrophagetropic viruses that enter via CC-chemokine receptor 5 (CCR5), despite binding to the same receptor as the virus. Furthermore, RANTES enhances the infectivity of HIV-1 pseudotyped with the envelope glycoprotein of murine leukemia virus or vesicular stomatitis virus, showing that the mechanism of enhancement is independent of the route of virus-cell fusion. The enhancing effects of RANTES are not mediated via CCR5 or other known chemokine receptors and are not mimicked by MIP-1alpha or MIP-1beta. The N-terminally modified derivative aminooxypentane RANTES (AOP-RANTES) efficiently inhibits HIV-1 infection via CCR5 but otherwise mimics RANTES by enhancing viral infectivity. There are two mechanisms of enhancement: one apparent when target cells are pretreated with RANTES (or AOP-RANTES) for several hours, and the other apparent when RANTES (or AOP-RANTES) is added during virus-cell absorption. We believe that the first mechanism is related to cellular activation by RANTES, whereas the second is an increase in virion attachment to target cells. PMID- 9847375 TI - Reovirus-induced apoptosis is preceded by increased cellular calpain activity and is blocked by calpain inhibitors. AB - The cellular pathways of apoptosis have not been fully characterized; however, calpain, a cytosolic calcium-activated cysteine protease, has been implicated in several forms of programmed cell death. Reoviruses induce apoptosis both in vitro and in vivo and serve as a model for studying virus-induced cell death. We investigated the potential role of calpain in reovirus-induced apoptosis in vitro by measuring calpain activity as well as evaluating the effects of calpain inhibitors. L929 cells were infected with reovirus type 3 Abney (T3A), and calpain activity, measured as cleavage of the fluorogenic calpain substrate Suc Leu-Leu-Val-Tyr-AMC, was monitored. There was a 1.6-fold increase in calpain activity in T3A-infected cells compared to mock-infected cells; this increase was completely inhibited by preincubation with calpain inhibitor I (N-acetyl-leucyl leucyl-norleucinal [aLLN]), an active-site inhibitor. Both aLLN and PD150606, a specific calpain inhibitor that interacts with the calcium-binding site, inhibited reovirus-induced apoptosis in L929 cells by 54 to 93%. Apoptosis induced by UV-inactivated reovirus was also reduced 65 to 69% by aLLN, indicating that inhibition of apoptosis by calpain inhibitors is independent of effects on viral replication. We conclude that calpain activation is a component of the regulatory cascade in reovirus-induced apoptosis. PMID- 9847376 TI - Sendai virus infection induces apoptosis through activation of caspase-8 (FLICE) and caspase-3 (CPP32). AB - Sendai virus (SV) infection and replication lead to a strong cytopathic effect with subsequent death of host cells. We now show that SV infection triggers an apoptotic program in target cells. Incubation of infected cells with the peptide inhibitor z-VAD-fmk abrogated SV-induced apoptosis, indicating that proteases of the caspase family were involved. Moreover, proteolytic activation of two distinct caspases, CPP32/caspase-3 and, as shown for the first time in virus infected cells, FLICE/caspase-8, could be detected. So far, activation of FLICE/caspase-8 has been described in apoptosis triggered by death receptors, including CD95 and tumor necrosis factor (TNF)-R1. In contrast, we could show that SV-induced apoptosis did not require TNF or CD95 ligand. We further found that apoptosis of infected cells did not influence the maturation and budding of SV progeny. In conclusion, SV-induced cell injury is mediated by CD95- and TNF-R1 independent activation of caspases, leading to the death of host cells without impairment of the viral life cycle. PMID- 9847377 TI - Cleavage of Poly(A)-binding protein by coxsackievirus 2A protease in vitro and in vivo: another mechanism for host protein synthesis shutoff? AB - Infection of cells by picornaviruses of the rhinovirus, aphthovirus, and enterovirus groups results in the shutoff of host protein synthesis but allows viral protein synthesis to proceed. Although considerable evidence suggests that this shutoff is mediated by the cleavage of eukaryotic translation initiation factor eIF4G by sequence-specific viral proteases (2A protease in the case of coxsackievirus), several experimental observations are at variance with this view. Thus, the cleavage of other cellular proteins could contribute to the shutoff of host protein synthesis and stimulation of viral protein synthesis. Recent evidence indicates that the highly conserved 70-kDa cytoplasmic poly(A) binding protein (PABP) participates directly in translation initiation. We have now found that PABP is also proteolytically cleaved during coxsackievirus infection of HeLa cells. The cleavage of PABP correlated better over time with the host translational shutoff and onset of viral protein synthesis than did the cleavage of eIF4G. In vitro experiments with purified rabbit PABP and recombinant human PABP as well as in vivo experiments with Xenopus oocytes and recombinant Xenopus PABP demonstrate that the cleavage is catalyzed by 2A protease directly. N- and C-terminal sequencing indicates that cleavage occurs uniquely in human PABP at 482VANTSTQTM downward arrowGPRPAAAAAA500, separating the four N-terminal RNA recognition motifs (80%) from the C-terminal homodimerization domain (20%). The N-terminal cleavage product of PABP is less efficient than full-length PABP in restoring translation to a PABP-dependent rabbit reticulocyte lysate translation system. These results suggest that the cleavage of PABP may be another mechanism by which picornaviruses alter the rate and spectrum of protein synthesis. PMID- 9847378 TI - Cleavage of poly(A)-binding protein by enterovirus proteases concurrent with inhibition of translation in vitro. AB - Many enteroviruses, members of the family Picornaviridae, cause a rapid and drastic inhibition of host cell protein synthesis during infection, a process referred to as host cell shutoff. Poliovirus, one of the best-studied enteroviruses, causes marked inhibition of host cell translation while preferentially allowing translation of its own genomic mRNA. An abundance of experimental evidence has accumulated to indicate that cleavage of an essential translation initiation factor, eIF4G, during infection is responsible at least in part for this shutoff. However, evidence from inhibitors of viral replication suggests that an additional event is necessary for the complete translational shutoff observed during productive infection. This report examines the effect of poliovirus infection on a recently characterized 3' end translational stimulatory protein, poly(A)-binding protein (PABP). PABP is involved in stimulating translation initiation in lower eukaryotes by its interaction with the poly(A) tail on mRNAs and has been proposed to facilitate 5'-end-3'-end interactions in the context of the closed-loop translational model. Here, we show that PABP is specifically degraded during poliovirus infection and that it is cleaved in vitro by both poliovirus 2A and 3C proteases and coxsackievirus B3 2A protease. Further, PABP cleavage by 2A protease is accompanied by concurrent loss of translational activity in an in vitro-translation assay. Similar loss of translational activity also occurs simultaneously with partial 3C protease mediated cleavage of PABP in translation assays. Further, PABP is not degraded during infections in the presence of guanidine-HCl, which blocks the complete development of host translation shutoff. These results provide preliminary evidence that cleavage of PABP may contribute to inhibition of host translation in infected HeLa cells, and they are consistent with the hypothesis that PABP plays a role in facilitating translation initiation in higher eukaryotes. PMID- 9847379 TI - Intrinsic human immunodeficiency virus type 1 resistance of hematopoietic stem cells despite coreceptor expression. AB - Interactions of human immunodeficiency virus type 1 (HIV-1) with hematopoietic stem cells may define restrictions on immune reconstitution following effective antiretroviral therapy and affect stem cell gene therapy strategies for AIDS. In the present study, we demonstrated mRNA and cell surface expression of HIV-1 receptors CD4 and the chemokine receptors CCR-5 and CXCR-4 in fractionated cells representing multiple stages of hematopoietic development. Chemokine receptor function was documented in subsets of cells by calcium flux in response to a cognate ligand. Productive infection by HIV-1 via these receptors was observed with the notable exception of stem cells, in which case the presence of CD4, CXCR 4, and CCR-5, as documented by single-cell analysis for expression and function, was insufficient for infection. Neither productive infection, transgene expression, nor virus entry was detectable following exposure of stem cells to either wild-type HIV-1 or lentivirus constructs pseudotyped in HIV-1 envelopes of macrophage-tropic, T-cell-tropic, or dualtropic specificity. Successful entry into stem cells of a vesicular stomatitis virus G protein-pseudotyped HIV-1 construct demonstrated that the resistance to HIV-1 was mediated at the level of virus-cell membrane fusion and entry. These data define the hematopoietic stem cell as a sanctuary cell which is resistant to HIV-1 infection by a mechanism independent of receptor and coreceptor expression that suggests a novel means of cellular protection from HIV-1. PMID- 9847380 TI - Phosphorylation of the human T-cell leukemia virus type 1 transactivator tax on adjacent serine residues is critical for tax activation. AB - The Tax transactivator protein of human T-cell leukemia virus type 1 (HTLV-1) plays a central role in the activation of viral gene expression. In addition, Tax is capable of activating the expression of specific cellular genes and is involved in the transformation of T-lymphocytes resulting in the development of adult T-cell leukemia. Tax is a phosphoprotein that colocalizes in nuclear bodies with RNA polymerase II, splicing complexes, and specific transcription factors including members of the ATF/CREB and NF-kappaB families. In this study, we identified adjacent serine residues at positions 300 and 301 in the carboxy terminus of Tax as the major sites for phosphorylation. Phosphorylation of at least one of these serine residues is required for Tax localization in nuclear bodies and for Tax-mediated activation of gene expression via both the ATF/CREB and NF-kappaB pathways. Introduction of amino acid substitutions which are phosphoserine mimetics at positions 300 and 301 restored the ability of a phosphorylation-defective Tax mutant to form nuclear bodies and to activate gene expression. These studies define sites for regulatory phosphorylation events in Tax which are critical for its ability to activate gene transcription. PMID- 9847382 TI - "Hidden" dUTPase sequence in human immunodeficiency virus type 1 gp120. AB - A coding region homologous to the sequence for essential eukaryotic enzyme dUTPase has been identified in different genomic regions of several viral lineages. Unlike the nonprimate lentiviruses (caprine arthritis- encephalitis virus, equine infectious anemia virus, feline immunodeficiency virus, and visna virus), where dUTPase is integrated into the pol coding region, this enzyme has never been demonstrated to be present in the primate lentivirus genomes (human immunodeficiency virus type 1 [HIV-1], HIV-2, or the related simian immunodeficiency virus). A novel approach allowed us to identify a weak but significant sequence similarity between HIV-1 gp120 and the human dUTPase. This finding was then extended to all of the primate lentivirus lineages. Together with the recently reported fragmentary structural similarity between the V3 loop region and the Escherichia coli dUTPase (P. D. Kwong, R. Wyatt, J. Robinson, R. W. Sweet, J. Sodroski, and W. A. Hendrickson, Nature 393:648-659, 1998), our results strongly suggest that an ancestral dUTPase gene has evolved into the present primate lentivirus CD4 and cytokine receptor interacting region of gp120. PMID- 9847381 TI - Nuclear magnetic resonance analysis of solution conformations in C4-V3 hybrid peptides derived from human immunodeficiency virus (HIV) type 1 gp120: relation to specificity of peptide-induced anti-HIV neutralizing antibodies. AB - Immunogenic peptides containing epitopes of the gp120 C4 and V3 regions from human immunodeficiency virus strains MN and EV91 have been studied by nuclear magnetic resonance and molecular modeling and used as immunogens in rhesus monkeys. The results, combined with those for other peptides, suggest a correlation between solution conformation and immunologic cross-reactivity. PMID- 9847383 TI - Human herpesvirus 6 latently infects early bone marrow progenitors in vivo. AB - We have studied the in vivo tropism of human herpesvirus 6 (HHV-6) for hemopoietic cells in patients with latent HHV-6 infection. Having used a variety of cell purification, molecular, cytogenetic, and immunocytochemical procedures, we report the first evidence that HHV-6 latently infects early bone marrow progenitor cells and that HHV-6 may be transmitted longitudinally to cells which differentiate along the committed pathways. PMID- 9847384 TI - Fine structure and morphogenesis of Borna disease virus. AB - Borna disease virus (BDV), a negative nonsegmented single-stranded RNA virus, has not been fully characterized morphologically. Here we present what is to our knowledge the first data on the fine ultrastructure and morphogenesis of BDV. The supernatant of MDCK cells persistently infected with BDV treated with n-butyrate contained many virus-like particles and more BDV-specific RNA than that of untreated samples. The particles were spherical, enveloped, and approximately 130 nm in diameter; had spikes 7 nm in length; and reacted with BDV p40 antibody. A thin nucleocapsid, 4 nm in width, was present peripherally in contrast to the thick nucleocapsid of hemagglutinating virus of Japan. The BDV particles reproduced by budding on the cell surface. PMID- 9847385 TI - Identical 371-base-pair deletion mutations in the LAT genes of herpes simplex virus type 1 McKrae and 17syn+ result in different in vivo reactivation phenotypes. AB - The herpes simplex virus type 1 (HSV-1) LAT gene is the only viral gene abundantly transcribed during latency. LAT null mutants created with strains McKrae and 17syn+ are impaired for both in vivo spontaneous and in vivo-induced reactivation. Thus, LAT is essential for efficient in vivo-induced and spontaneous reactivation. Different investigators have studied two LAT mutants containing a StyI-StyI region deletion corresponding to LAT nucleotides 76 to 447. One mutant, dLAT371 (parent strain, McKrae), had parental high frequencies of spontaneous reactivation. In vivo-induced reactivation was not examined. The other mutant, 17DeltaSty (parent strain, 17syn+), had parental frequencies of in vitro reactivation following cocultivation of explanted ganglia but reduced frequencies of in vivo-induced reactivation. Spontaneous reactivation frequency was not reported for 17DeltaSty. These combined results suggested the possibility that in vivo spontaneous reactivation and in vivo-induced reactivation may map to different regions within the LAT domain. We now report that dLAT371 has in vivo induced reactivation frequencies of the parent and that 17DeltaSty has reduced frequencies of in vivo spontaneous reactivation. Thus, dLAT371 demonstrated the parental phenotype for both in vivo spontaneous and -induced reactivation while the apparently identical 17DeltaSty was impaired for both in vivo spontaneous and -induced reactivation. These results suggest that one or more differences between the genetic backgrounds of McKrae and 17syn+ result in different in vivo reactivation phenotypes of otherwise identical deletion mutations and that McKrae may have compensating sequences sufficient to overcome the loss of the StyI-StyI region of the LAT transcript. PMID- 9847387 TI - Lack of effect of cytoplasmic tail truncations on human immunodeficiency virus type 2 ROD env particle release activity. AB - In addition to its role in receptor binding, the envelope glycoprotein of certain human immunodeficiency virus type 2 (HIV-2) isolates, including ROD10, exhibits a biological activity that enhances the release of HIV-2, HIV-1, and simian immunodeficiency virus particles from infected cells. The present study aims at better defining the functional domains involved in this biological activity. To this end, we have characterized the envelope protein of the ROD14 isolate of HIV 2, which, despite 95% homology with the ROD10 envelope at the amino acid level, is unable to enhance viral particle release. Site-directed mutagenesis showed that the presence of a truncation in the cytoplasmic tail of the ROD14 envelope was not responsible for the lack of activity, as previously reported for the HIV 2 ST isolate (G. D. Ritter, Jr., G. Yamshchikov, S. J. Cohen, and M. J. Mulligan, J. Virol. 70:2669-2673, 1996). Similarly, several modifications of the length of the ROD10 envelope cytoplasmic tail did not impair its ability to enhance particle release, suggesting that, in the case of the HIV-2 ROD isolate, particle release activity is not regulated by the length of the cytoplasmic tail. PMID- 9847386 TI - Polypyrimidine tract-binding protein binds to the leader RNA of mouse hepatitis virus and serves as a regulator of viral transcription. AB - A cellular protein, previously described as p55, binds specifically to the plus strand of the mouse hepatitis virus (MHV) leader RNA. We have purified this protein and determined by partial peptide sequencing that it is polypyrimidine tract-binding protein (PTB) (also known as heterogeneous nuclear ribonucleoprotein [hnRNP] I), a nuclear protein which shuttles between the nucleus and cytoplasm. PTB plays a role in the regulation of alternative splicing of pre-mRNAs in normal cells and translation of several viruses. By UV cross linking and immunoprecipitation studies using cellular extracts and a recombinant PTB, we have established that PTB binds to the MHV plus-strand leader RNA specifically. Deletion analyses of the leader RNA mapped the PTB-binding site to the UCUAA pentanucleotide repeats. Using a defective-interfering RNA reporter system, we have further shown that the PTB-binding site in the leader RNA is critical for MHV RNA synthesis. This and our previous study (H.-P. Li, X. Zhang, R. Duncan, L. Comai, and M. M. C. Lai, Proc. Natl. Acad. Sci. USA 94:9544-9549, 1997) combined thus show that two cellular hnRNPs, PTB and hnRNP A1, bind to the transcription-regulatory sequences of MHV RNA and may participate in its transcription. PMID- 9847388 TI - New mouse model for dengue virus vaccine testing. AB - Several dengue (DEN) virus vaccines are in development; however, the lack of a reliable small animal model in which to test them is a major obstacle. Because evidence suggests that interferon (IFN) is involved in the human anti-DEN virus response, we tested mice deficient in their IFN functions as potential models. Intraperitoneally administered mouse-adapted DEN 2 virus was uniformly lethal in AG129 mice (which lack alpha/beta IFN and gamma IFN receptor genes), regardless of age. Immunized mice were protected from virus challenge, and survival times increased following passive transfer of anti-DEN polyclonal antibody. These results demonstrate that AG129 mice are a promising small animal model for DEN virus vaccine trials. PMID- 9847389 TI - Characterization of a natural mutation in an antigenic site on the fusion protein of measles virus that is involved in neutralization. AB - Although measles virus is an antigenically monotypic virus, nucleotide sequence analysis of the hemagglutinin and nucleoprotein genes has permitted the differentiation of a number of genotypes. In contrast, the fusion (F) protein is highly conserved; only three amino acid changes have been reported over a 40-year period. We have isolated a measles virus strain which did not react with an anti F monoclonal antibody (MAb) which we had previously shown to be directed against a dominant antigenic site. This virus strain, Lys-1, had seven amino acid changes compared with the Edmonston strain. We have shown that a single amino acid at position 73 is responsible for its nonreactivity with the anti-F MAb. With the same MAb, antibody-resistant mutants were prepared from the vaccine strain. A single amino acid change at position 73 (R-->W) was observed. The possibility of selecting measles virus variants in vaccinated populations is discussed. PMID- 9847390 TI - Down regulation of gene expression by the vaccinia virus D10 protein. AB - Vaccinia virus genes are expressed in a sequential fashion, suggesting a role for negative as well as positive regulatory mechanisms. A potential down regulator of gene expression was mapped by transfection assays to vaccinia virus open reading frame D10, which encodes a protein with no previously known function. Inhibition was independent of the promoter type used for the reporter gene, indicating that the mechanism did not involve promoter sequence recognition. The inhibition was overcome, however, when the open reading frame of the reporter gene was preceded by the encephalomyocarditis virus internal ribosome entry site, which excludes the possibility of nonspecific metabolic or other antiviral effects and suggests that capped mRNAs or cap-dependent translation might be the target of the D10 product. The inducible overexpression of the D10 gene by a recombinant vaccinia virus severely inhibited viral protein synthesis, decreased the steady-state level of viral late mRNA, and blocked the formation of infectious virus. PMID- 9847391 TI - Decay kinetics of human immunodeficiency virus-specific effector cytotoxic T lymphocytes after combination antiretroviral therapy. AB - Little is known of the changes in human immunodeficiency virus type 1 (HIV-1) specific effector cytotoxic T lymphocytes (CTL) after potent antiretroviral therapy. Using HLA/peptide tetrameric complexes, we show that after starting treatment, there are early rapid fluctuations in the HIV-1-specific CTL response which last 1 to 2 weeks. These fluctuations are followed by an exponential decay (median half-life, 45 days) of HIV-1-specific CTL which continues while viremia remains undetectable. These data have implications for the immunological control of drug-resistant virus. PMID- 9847392 TI - An attenuated variant of the GDVII strain of Theiler's virus does not persist and does not infect the white matter of the central nervous system. AB - The DA strain of Theiler's virus causes a persistent and demyelinating infection of the white matter of spinal cord, whereas the GDVII strain causes a fatal gray matter encephalomyelitis. Studies with recombinant viruses showed that this difference in phenotype is controlled mainly by the capsid. However, conflicting results regarding the existence of determinants of persistence in the capsid of the GDVII strain have been published. Here we show that a GDVII virus whose neurovirulence has been attenuated by an insertion in the 5' noncoding region does not persist in the central nervous systems of mice. Furthermore, this virus infects the gray matter efficiently, but not the white matter. These results confirm the absence of determinants of persistence in the GDVII capsid. They suggest that the DA capsid controls persistence by allowing the virus to infect cells in the white matter of the spinal cord. PMID- 9847393 TI - RNA replication for the paramyxovirus simian virus 5 requires an internal repeated (CGNNNN) sequence motif. AB - A functional RNA replication promoter for the paramyxovirus simian virus 5 (SV5) requires two essential and discontinuous elements: 19 bases at the 3' terminus (conserved region I) and an 18-base internal region (conserved region II [CRII]) that is contained within the coding region of the L protein gene. A reverse genetics system was used to determine the sequence requirements for the internal CRII element to function in RNA replication. A series of copyback defective interfering (DI) RNA analogs were constructed to contain point mutations in the 18 nucleotides composing CRII, and their relative replication levels were analyzed. The results indicated that SV5 DI RNA replication was reduced by substitutions for two CG dinucleotides, which in the nucleocapsid template are in the first two positions of the first two hexamers of CRII nucleotides. Substitutions for other bases within CRII did not reduce RNA synthesis. Thus, two consecutive 5'-CGNNNN-3' hexamers form an important sequence in the SV5 CRII promoter element. The position of the CG dinucleotide within the SV5 leader and antitrailer promoters was highly conserved among other members of the Rubulavirus genus, but this motif differed significantly in both sequence and position from that previously identified for Sendai virus. The possible roles of the CRII internal promoter element in paramyxovirus RNA replication are discussed. PMID- 9847394 TI - Herpes simplex virus genome isomerization: origins of adjacent long segments in concatemeric viral DNA. AB - Herpes simplex virus type 1 DNA isomerization was studied by using a viral mutant, 5B8, lacking the unique SpeI site of its parent, SC16. In coinfected cells, SC16 genomic long segments flanked 5B8 genomes in all possible orientations with similar frequencies. Thus, recombination between progeny of different replication templates is sufficient to explain genomic isomerization. PMID- 9847395 TI - Genetic selection of poliovirus 2Apro-binding peptides. AB - The yeast two-hybrid system has been used to identify mammalian clones that interact with poliovirus 2A proteinase (2Apro). Eight clones which encode previously unidentified human proteins were selected from a HeLa cell cDNA expression library. In addition, five clones encoding short peptides that interact with poliovirus 2Apro were also identified. The lengths of these peptides range from 6 to 30 amino acids, but all of them contain the Leu-X-Thr-Z motif (X represents any amino acid; Z represents a hydrophobic residue). This sequence is invariably located just at the carboxy terminus of each peptide. This approach raises the possibility of designing substrate analogue inhibitors of 2Apro. Thus, two nonhydrolyzable peptides containing the Leu-X-Thr-Z motif prevented cleavage of eukaryotic initiation factor 4G by poliovirus 2Apro in vitro. A more general method for identifying peptides with antiproteinase activity is discussed. PMID- 9847396 TI - Molecular characterization of a bovine enteric calicivirus: relationship to the Norwalk-like viruses. AB - Jena virus (JV) is a noncultivatable bovine enteric calicivirus associated with diarrhea in calves and was first described in Jena, Germany. The virus was serially passaged 11 times in colostrum-deprived newborn calves and caused diarrheal disease symptoms at each passage. The complete JV genome sequence was determined by using cDNA made from partially purified virus obtained from a single stool sample. JV has a positive-sense single-stranded RNA genome which is 7,338 nucleotides in length, excluding the poly(A) tail. JV genome organization is similar to that of the human Norwalk-like viruses (NLVs), with three separate open reading frames (ORFs) and a 24-nucleotide sequence motif located at the 5' terminus of the genome and at the start of ORF 2. The polyprotein (ORF 1) consists of 1,680 amino acids and has the characteristic 2C helicase, 3C protease, and 3D RNA polymerase motifs also found in the NLVs. However, comparison of the N-terminal 100 amino acids of the JV polyprotein with those of the group 1 and group 2 NLVs showed a considerable divergence in sequence. The capsid protein (ORF 2) at 519 amino acids is smaller than that of all other caliciviruses. JV ORF 2 was translated in vitro to produce a 55-kDa protein that reacted with postinfection serum but not preinfection serum. Phylogenetic studies based on partial RNA polymerase sequences indicate that within the Caliciviridae JV is most closely related to the group 1 NLVs. PMID- 9847398 TI - Feline calicivirus capsid protein expression and capsid assembly in cultured feline cells. AB - The capsid protein of feline calicivirus (FCV) was expressed by using plasmids containing cytomegalovirus, simian virus 40, or T7 promoters. The strongest expression was achieved with the T7 promoter and coinfection with vaccinia virus expressing the T7 RNA polymerase (MVA/T7pol). The FCV precursor capsid protein was processed to the mature-size protein, and these proteins were assembled in to virus-like particles. PMID- 9847397 TI - Transcriptional activation signals found in the Epstein-Barr virus (EBV) latency C promoter are conserved in the latency C promoter sequences from baboon and Rhesus monkey EBV-like lymphocryptoviruses (cercopithicine herpesviruses 12 and 15). AB - The Epstein-Barr virus (EBV) EBNA2 protein is a transcriptional activator that controls viral latent gene expression and is essential for EBV-driven B-cell immortalization. EBNA2 is expressed from the viral C promoter (Cp) and regulates its own expression by activating Cp through interaction with the cellular DNA binding protein CBF1. Through regulation of Cp and EBNA2 expression, EBV controls the pattern of latent protein expression and the type of latency established. To gain further insight into the important regulatory elements that modulate Cp usage, we isolated and sequenced the Cp regions corresponding to nucleotides 10251 to 11479 of the EBV genome (-1079 to +144 relative to the transcription initiation site) from the EBV-like lymphocryptoviruses found in baboons (herpesvirus papio; HVP) and Rhesus macaques (RhEBV). Sequence comparison of the approximately 1,230-bp Cp regions from these primate viruses revealed that EBV and HVP Cp sequences are 64% conserved, EBV and RhEBV Cp sequences are 66% conserved, and HVP and RhEBV Cp sequences are 65% conserved relative to each other. Approximately 50% of the residues are conserved among all three sequences, yet all three viruses have retained response elements for glucocorticoids, two positionally conserved CCAAT boxes, and positionally conserved TATA boxes. The putative EBNA2 100-bp enhancers within these promoters contain 54 conserved residues, and the binding sites for CBF1 and CBF2 are well conserved. Cp usage in the HVP- and RhEBV-transformed cell lines was detected by S1 nuclease protection analysis. Transient-transfection analysis showed that promoters of both HVP and RhEBV are responsive to EBNA2 and that they bind CBF1 and CBF2 in gel mobility shift assays. These results suggest that similar mechanisms for regulation of latent gene expression are conserved among the EBV-related lymphocryptoviruses found in nonhuman primates. PMID- 9847399 TI - Binding specificity of protein phosphatase 2A core enzyme for regulatory B subunits and T antigens. AB - The core enzyme of protein phosphatase 2A is composed of a regulatory subunit A and a catalytic subunit C. It is controlled by three types of regulatory B subunits (B, B', and B") and by tumor (T) antigens, which are unrelated by sequence but bind to overlapping regions on the A subunit. To find out whether the different B subunits and T antigens bind to identical or distinct amino acids of the A subunit, mutants were generated and their abilities to bind B subunits and T antigens were tested. We found that some amino acids are involved in the binding of all types of B subunits, whereas others are specifically involved in the binding of one or two types of B subunits. T-antigen-binding specificity does not correlate with that of a particular type of B subunit. PMID- 9847400 TI - Truncated particles produced in fish surviving infectious hematopoietic necrosis virus infection: mediators of persistence? AB - Infectious hematopoietic necrosis virus (IHNV) is a rhabdovirus that produces an acute, lethal infection in rainbow trout (Oncorhynchus mykiss). Fish that survive infection cease to produce detectable infectious virus at approximately 46 days after infection, yet there is evidence that survivor fish continue to harbor virus particles (B. S. Drolet, P. P. Chiou, J. Heidel, and J. C. Leong, J. Virol. 69:2140-2147, 1995). In an effort to determine the biological function of these particles, the kidneys and livers from IHNV survivors were harvested and divided into samples for nested reverse transcriptase PCR analysis and explant culture. Sequences for the IHNV nucleoprotein and polymerase genes were detected in 50 and 89%, respectively, of the organs from survivor fish. When explant tissue cultures were infected with purified standard IHNV, the liver tissues from survivor fish produced up to 10-fold less virus than naive control fish liver tissues. In addition, immunosorbent electron microscopy analysis of the supernatant media from the cultured explants of survivor fish revealed truncated particles, whereas the control tissue supernatants contained only standard viral particles. These results suggest that the truncated IHNV particles observed in persistently infected fish are defective interfering particles that may mediate virus persistence. PMID- 9847401 TI - Constrained evolution of human immunodeficiency virus type 1 protease during sequential therapy with two distinct protease inhibitors. AB - Human immunodeficiency virus type 1 (HIV-1) variants that have developed protease (PR) inhibitor resistance most often display cross-resistance to several molecules within this class of antiretroviral agents. The clinical benefit of the switch to a second PR inhibitor in the presence of such resistant viruses may be questionable. We have examined the evolution of HIV-1 PR genotypes and phenotypes in individuals having failed sequential treatment with two distinct PR inhibitors: saquinavir (SQV) followed by indinavir (IDV). In viruses where typical SQV resistance mutations were detected before the change to IDV, the corresponding mutations were maintained under IDV, while few additional mutations emerged. In viruses where no SQV resistance mutations were detected before the switch to IDV, typical SQV resistance profiles emerged following the introduction of IDV. We conclude that following suboptimal exposure to a first PR inhibitor, the introduction of a second molecule of this class can lead to rapid selection of cross-resistant virus variants that may not be detectable by current genotyping methods at the time of the inhibitor switch. Viruses committed to resistance to the first inhibitor appear to bear the "imprint" of this initial selection and can further adapt to the selective pressure exerted by the second inhibitor following a pathway that preserves most of the initially selected mutations. PMID- 9847402 TI - Rapid clearance of simian immunodeficiency virus particles from plasma of rhesus macaques. AB - Perturbation of the equilibrium between human immunodeficiency virus type 1 (HIV 1) and the infected host by administering antiretroviral agents has revealed the rapid turnover of both viral particles and productively infected cells. In this study, we used the infusion of simian immunodeficiency virus (SIV) particles into rhesus macaques to obtain a more accurate estimate of viral clearance in vivo. Consistently, exogenously infused virions were cleared from plasma with an extremely short half-life, on the order of minutes (a mean of 3.3 min). This new estimate is approximately 100-fold lower than the upper bound of 6 h previously reported for HIV-1 in infected humans. In select animals, multiple tissues were collected at the completion of each experiment to track the potential sites of virion clearance. Detectable levels of SIV RNA were found in lymph nodes, spleen, lungs, and liver, but not in other tissues examined. However, only approximately 1 to 10% or less of the infused virions were accounted for by the thorough tissue sampling, indicating that the vast majority of the infused particles must have been degraded over a short period of time. Should the rapid clearance of virions described here be applicable to infected patients, then HIV-1 production and thus the number of productively infected CD4(+) T lymphocytes or the viral burst size must be proportionally higher than previous minimal estimates. PMID- 9847403 TI - The phylogeny of glyceraldehyde-3-phosphate dehydrogenase indicates lateral gene transfer from cryptomonads to dinoflagellates. AB - Sequence analysis of two nuclear-encoded glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes isolated from the dinoflagellate Gonyaulax polyedra distinguishes them as cytosolic and chloroplastic forms of the enzyme. Distance analysis of the cytosolic sequence shows the Gonyaulax gene branching early within the cytosolic clade, consistent with other analyses. However, the plastid sequence forms a monophyletic group with the plastid isoforms of cryptomonads, within an otherwise cytosolic clade, distinct from all other plastid GAPDHs. This is attributed to lateral gene transfer from an ancestral cryptomonad to a dinoflagellate, providing the first example of genetic exchange accompanying symbiotic associations between the two, which are common in present day cells. PMID- 9847404 TI - Sponge Pax cDNA related to Pax-2/5/8 and ancient gene duplications in the Pax family. AB - Members of the Pax gene family encode transcription factors containing a DNA binding paired domain which is involved in developmental control and the formation of the central nervous system (CNS). The family members are classified into six classes or subfamilies, depending on the presence or absence of paired type homeobox and octapeptide. To obtain rough estimates of times when the different classes of the Pax family diverged by gene duplication, we cloned and sequenced a Pax-related cDNA, sPax-2/5/8, from Ephydatia fluviatilis, a freshwater sponge, which encodes a paired-type homeobox and an octapeptide, in addition to a paired domain. A phylogenetic tree based on the paired domain sequences suggest that sPax-2/5/8 is a homologue of vertebrate Pax-2/5/8. It was also suggested that the majority of gene duplications that gave rise to distinct classes has been completed in the very early evolution of animals before the parazoan-eumetazoan split. Long after the ancient gene duplications, further gene duplications that gave rise to members in each subfamily occurred on the chordate lineages and completed before the fish-tetrapod split. This suggests that the major classes of the Pax genes involved in the formation of CNS characteristic of triploblasts had already existed long before the Cambrian explosion of triploblasts, and there is no direct link between the creation of new genes with novel functions and the Cambrian explosion. The pattern of gene diversification found in the Pax family is similar to those in five gene families involved in the signal transduction analyzed by us. Furthermore, the evolutionary rates of the Pax proteins have been shown to decrease with increasing organismal complexity during animal evolution. PMID- 9847405 TI - Evolutionary relationship between translation initiation factor eIF-2gamma and selenocysteine-specific elongation factor SELB: change of function in translation factors. AB - Eubacterial and eukaryotic translation initiation systems have very little in common, and therefore the evolutionary events that gave rise to these two disparate systems are difficult to ascertain. One common feature is the presence of initiation, elongation, and release factors belonging to a large GTPase superfamily. One of these initiation factors, the gamma subunit of initiation factor 2 (eIF-2gamma), is found only in eukaryotes and archaebacteria. We have sequenced eIF-2gamma gene fragments from representative diplomonads, parabasalia, and microsporidia and used these new sequences together with new archaebacterial homologues to examine the phylogenetic position of eIF-2gamma within the GTPase superfamily. The archaebacterial and eukaryotic eIF-2gamma proteins are found to be very closely related, and are in turn related to SELB, the selenocysteine specific elongation factor from eubacteria. The overall topology of the GTPase tree further suggests that the eIF-2gamma/SELB group may represent an ancient subfamily of GTPases that diverged prior to the last common ancestor of extant life. PMID- 9847406 TI - Identification of a TRAF (TNF receptor-associated factor) gene in Caenorhabditis elegans. AB - Many members of the tumor necrosis factor (TNF) receptor superfamily and the interleukin-1 (IL-1) receptor engage intracellular signaling pathways including the nuclear factor kappaB (NF-kappaB)-, c-jun N-terminal kinase (JNK)-, and extracellular signal-regulated kinase (ERK) pathways by direct or indirect interaction with TNF receptor-associated factor (TRAF) molecules. To date, six mammalian members of the TRAF family have been identified. Searching public databases with a sequence pattern comprising 19 conserved amino acid residues derived from the carboxyl-terminal part of the TRAF homology domain, we found significant sequence homologies to a stretch of genomic DNA from Caenorhabditis elegans which encodes 1 of 12 exons of a putative protein. The sequence of this putative protein shows up to 29% sequence identity to the mammalian TRAFs and is therefore designated C. elegans TRAF (CeTRAF). The CeTRAF molecule has an amino terminal RING finger motif followed by four zinc finger structures and a carboxyl terminal TRAF domain, a composition which is also found in most of the mammalian TRAFs. Reverse transcription-PCR and sequencing analysis of the respective amplicon clearly demonstrates that CeTRAF is in fact transcribed in C. elegans. The existence of a member of the TRAF family in C. elegans provides strong evidence for evolutionary conserved pathways linking cell surface receptors to activation of JNK, ERK, and NF-kappaB. PMID- 9847407 TI - Identification of evolutionarily invariant sequences in the protein C gene promoter. AB - Recent studies on human protein C gene expression have revealed the presence of three transcription factor binding sites in close proximity to the transcription start site. Binding sites for the liver-enriched hepatocyte nuclear factors 1 and 3 (HNF-1 and HNF-3, respectively) are located immediately upstream of the transcription start site, whereas just downstream of the start site a presently unidentified transcription factor may bind. To identify other candidate transcription factor binding sites in the protein C promoter, we studied the promoter sequence identity in a number of evolutionarily close and more distant species: Gorilla gorilla, Pongo pygmaeus, Pan troglodytes, Homo sapiens, Cebus apella, Macaca mulatta, Callithrix jacchus, Papio hamadryas, Macaca fascicularis, and Rattus norvegicus. This analysis showed that a high degree of identity (78%) exists among the different primates. Comparison of the primate consensus sequence with the Rattus norvegicus protein C promoter sequence revealed the presence of seven identical regions (I to VII). Two of these regions overlap with established regulatory sequences for HNF-3 and HNF-1 (region VI) and for PCE-1 (region VII), respectively. The functional importance and the transcription factors that may bind to the other five identical regions are now to be determined. PMID- 9847408 TI - Interelement selection in the regulatory region of the copia retrotransposon. AB - We report the results of an analysis of naturally occurring cis-regulatory variation within and between two families of the copia Drosophila long terminal repeat (LTR) retrotransposon. The copia 5' LTR and adjacent untranslated leader region (ULR) consists of a number of well-characterized sequence motifs which play a role in regulating expression of the element. In order to understand the evolutionary forces which may be responsible for generating and maintaining copia regulatory sequence variation, we have quantified levels of naturally occurring copia LTR-ULR nucleotide variation and subjected the data to a series of tests of neutrality. Our analysis indicates that the copia LTR-ULR has been subject to negative purifying selection within families and positive adaptive selection between families. We discuss these findings with respect to the regulatory evolution of retrotransposons and the phenomenon of interelement selection. PMID- 9847409 TI - The BC200 RNA gene and its neural expression are conserved in Anthropoidea (Primates). AB - The gene encoding BC200 RNA arose from a monomeric Alu element. Subsequently, the RNA had been recruited or exapted into a function of the nervous system. Here we confirm the presence of the BC200 gene in several primate species among the Anthropoidea. The period following the divergence of New World monkeys and Old World monkeys from their common ancestor is characterized by a significantly higher substitution rate in the examined 5' flanking region than in the BC200 RNA coding region itself. Furthermore, the conservation of CpG dimers in the RNA coding region (200 bp) is drastically increased compared to the 5' flanking region (approximately 400 bp) over all 12 species examined. Finally, the brain specific expression pattern of BC200 RNA and its presence as a ribonucleoprotein particle (RNP) are conserved in Old World and New World monkeys. Our studies indicate that the gene encoding BC200 RNA was created at least 35-55 million years ago and its presence, mode of expression, and association with protein(s) as an RNP are under selective pressure. PMID- 9847410 TI - Problems with parsimony in sequences of biased base composition. AB - Parsimony is commonly used to infer the direction of substitution and mutation. However, it is known that parsimony is biased when the base composition of the DNA sequence is skewed. Here I quantify this effect for several simple cases. The analysis demonstrates that parsimony can be misleading even when levels of sequence divergence are as low as 10%; parsimony incorrectly infers an excess of common to rare changes. Caution must therefore be excercised in the use of parsimony. PMID- 9847411 TI - Base composition skews, replication orientation, and gene orientation in 12 prokaryote genomes. AB - Variation in GC content, GC skew and AT skew along genomic regions was examined at third codon positions in completely sequenced prokaryotes. Eight out of nine eubacteria studied show GC and AT skews that change sign at the origin of replication. The leading strand in DNA replication is G-T rich at codon position 3 in six eubacteria, but C-T rich in two Mycoplasma species. In M. genitalium the AT and GC skews are symmetrical around the origin and terminus of replication, whereas its GC content variation has been shown to have a centre of symmetry elsewhere in the genome. Borrelia burgdorferi and Treponema pallidum show extraordinary extents of base composition skew correlated with direction of DNA replication. Base composition skews measured at third codon positions probably reflect mutational biases, whereas those measured over all bases in a sequence (or at codon positions 1 and 2) can be strongly affected by protein considerations due to the tendency in some bacteria for genes to be transcribed in the same direction that they are replicated. Consequently in some species the direction of skew for total genomic DNA is opposite to that for codon position 3. PMID- 9847412 TI - Sequence analysis of the mitochondrial genome of Sarcophyton glaucum: conserved gene order among octocorals. AB - The nucleotide sequence for an 11,715-bp segment of the mitochondrial genome of the octocoral Sarcophyton glaucum is presented, completing the analysis of the entire genome for this anthozoan member of the phylum Cnidaria. The genome contained the same 13 protein-coding and 2 ribosomal RNA genes as in other animals. However, it also included an unusual mismatch repair gene homologue reported previously and codes for only a single tRNA gene. Intermediate in length compared to two other cnidarians (17,443 and 18,911 bp), this organellar genome contained the smallest amount of noncoding DNA (428, compared to 1283 and 781 nt, respectively), making it the most compact one found for the phylum to date. The mitochondrial genes of S. glaucum exhibited an identical arrangement to that found in another octocoral, Renilla kolikeri, with five protein-coding genes in the same order as has been found in insect and vertebrate mitochondrial genomes. Although gene order appears to be highly conserved among octocorals, compared to the hexacoral, Metridium senile, few similarities were found. Like other metazoan mitochondrial genomes, the A + T composition was elevated and a general bias against codons ending in G or C was observed. However, an exception to this was the infrequent use of TGA compared to TGG to code for tryptophan. This divergent codon bias is unusual but appears to be a conserved feature among two rather distantly related anthozoans. PMID- 9847413 TI - Complete mitochondrial genome of a neotropical fruit bat, Artibeus jamaicensis, and a new hypothesis of the relationships of bats to other eutherian mammals. AB - The complete mitochondrial genome was obtained from a microchiropteran bat, Artibeus jamaicensis. The presumptive amino acid sequence for the protein-coding genes was compared with predicted amino acid sequences from several representatives of other mammalian orders. Data were analyzed using maximum parsimony, maximum likelihood, and neighbor joining. All analyses placed bats as the sister group of carnivores, perissodactyls, artiodactyls, and cetaceans (e.g., 100% bootstrap value with both maximum parsimony and neighbor joining). The data strongly support a new hypothesis about the origin of bats, specifically a bat/ferungulate grouping. None of the analyses supported the superorder Archonta (bats, flying lemurs, primates, and tree shrews). Our hypothesis regarding the relationship of bats to other eutherian mammals is concordant with previous molecular studies and contrasts with hypotheses based solely on morphological criteria and an incomplete fossil record. The A. jamaicensis mitochondrial DNA control region has a complex pattern of tandem repeats that differs from previously reported chiropteran control regions. PMID- 9847414 TI - Molecular timing of primate divergences as estimated by two nonprimate calibration points. AB - The complete mitochondrial DNA (mtDNA) molecule of the hamadryas baboon, Papio hamadryas, was sequenced and included in a molecular analysis of 24 complete mammalian mtDNAs. The particular aim of the study was to time the divergence between Cercopithecoidea and Hominoidea. That divergence, set at 30 million years before present (MYBP) was a fundamental reference for the original proposal of recent hominoid divergences, according to which the split among gorilla, chimpanzee, and Homo took place 5 MYBP. In the present study the validity of the postulated 30 MYBP dating of the Cercopithecoidea/Hominoidea divergence was examined by applying two independent nonprimate molecular references, the divergence between artiodactyls and cetaceans set at 60 MYBP and that between Equidae and Rhinocerotidae set at 50 MYBP. After calibration for differences in evolutionary rates, application of the two references suggested that the Cercopithecoidea/Hominoidea divergence took place >50 MYBP. Consistent with the marked shift in the dating of the Cercopithecoidea/Hominoidea split, all hominoid divergences receive a much earlier dating. Thus the estimated date of the divergence between Pan (chimpanzee) and Homo is 10-13 MYBP and that between Gorilla and the Pan/Homo linage approximately 17 MYBP. The same datings were obtained in an analysis of clocklike evolving genes. The findings show that recalculation is necessary of all molecular datings based directly or indirectly on a Cercopithecoidea/Hominoidea split 30 MYBP. PMID- 9847415 TI - Comparison and evolutionary analysis of the glycosomal glyceraldehyde-3-phosphate dehydrogenase from different Kinetoplastida. AB - In this work, we present the sequences and a comparison of the glycosomal GAPDHs from a number of Kinetoplastida. The complete gene sequences have been determined for some species (Crithidia fasciculata, Herpetomonas samuelpessoai, Leptomonas seymouri, and Phytomonas sp), whereas for other species (Trypanosoma brucei gambiense, Trypanosoma congolense, Trypanosoma vivax, and Leishmania major), only partial sequences have been obtained by PCR amplification. The structure of all available glycosomal GAPDH genes was analyzed in detail. Considerable variations were observed in both their nucleotide composition and their codon usage. The GC content varies between 64.4% in L. seymouri and 49.5% in the previously sequenced GAPDH gene from Trypanoplasma borreli. A highly biased codon usage was found in C. fasciculata, with only 34 triplets used, whereas in T. borreli 57 codons were employed. No obvious correlation could be observed between the codon usage and either the nucleotide composition or the level of gene expression. The glycosomal GAPDH is a very well-conserved enzyme. The maximal overall difference observed in the amino acid sequences is only 25%. Specific insertions and extensions are retained in all sequences. The residues involved in catalysis, substrate, and inorganic phosphate binding are fully conserved, whereas some variability is observed in the cofactor-binding pocket. The implications of these data for the design of new trypanocidal drugs targeted against GAPDH are discussed. All available gene and amino acid sequences of glycosomal GAPDHs were used for a phylogenetic analysis. The division of the Kinetoplastida into two suborders, Bodonina and Trypanosomatina, was well supported. Within the letter group, the Trypanosoma species appeared to be monophyletic, whereas the other trypanosomatids form a second clade. PMID- 9847416 TI - The pyrophosphate-dependent phosphofructokinase of the protist, Trichomonas vaginalis, and the evolutionary relationships of protist phosphofructokinases. AB - The pyrophosphate-dependent phosphofructokinase (PPi-PFK) of the amitochondriate protist Trichomonas vaginalis has been purified. The enzyme is a homotetramer of about 50 kDa subunits and is not subject to allosteric regulation. The protein was fragmented and a number of peptides were sequenced. Based on this information a PCR product was obtained from T. vaginalis gDNA and used to isolate corresponding cDNA and gDNA clones. Southern analysis indicated the presence of five genes. One open reading frame (ORF) was completely sequenced and for two others the 5' half of the gene was determined. The sequences were highly similar. The complete ORF corresponded to a polypeptide of about 46 kDa. All the peptide sequences obtained were present in the derived sequences. The complete ORF was highly similar to that of other PFKs, primarily in its amino-terminal half. The T. vaginalis enzyme was most similar to PPi-PFK of the mitochondriate heterolobosean, Naegleria fowleri. Most of the residues shown or assumed to be involved in substrate binding in other PPi-PFKs were conserved in the T. vaginalis enzyme. Direct comparison and phylogenetic reconstruction revealed a significant divergence among PPi-PFKs and related enzymes, which can be assigned to at least four distantly related groups, three of which contain enzymes of protists. The separation of these groups is supported with a high percentage of bootstrap proportions. The short T. vaginalis PFK shares a most recent common ancestor with the enzyme from N. fowleri. This pair is clearly separated from a group comprising the long (>60-kDa) enzymes from Giardia lamblia, Entamoeba histolytica pfk2, the spirochaetes Borrelia burgdorferi and Trepomena pallidum, as well as the alpha- and beta-subunits of plant PPi-PFKs. The third group ("X") containing protist sequences includes the glycosomal ATP-PFK of Trypanosoma brucei, E. histolytica pfk1, and a second sequence from B. burgdorferi. The fourth group ("Y") comprises cyanobacterial and high-G + C, Gram-positive eubacterial sequences. The well-studied PPi-PFK of Propionibacterium freudenreichii is highly divergent and cannot be assigned to any of these groups. These four groups are well separated from typical ATP-PFKs, the phylogenetic analysis of which confirmed relationships established earlier. These findings indicate a complex history of a key step of glycolysis in protists with several early gene duplications and possible horizontal gene transfers. PMID- 9847417 TI - Molecular phylogeny of metazoan intermediate filament proteins. AB - We have cloned cytoplasmic intermediate filament (IF) proteins from a large number of invertebrate phyla using cDNA probes, the monoclonal antibody IFA, peptide sequence information, and various RT-PCR procedures. Novel IF protein sequences reported here include the urochordata and nine protostomic phyla, i.e., Annelida, Brachiopoda, Chaetognatha, Echiura, Nematomorpha, Nemertea, Platyhelminthes, Phoronida, and Sipuncula. Taken together with the wealth of data on IF proteins of vertebrates and the results on IF proteins of Cephalochordata, Mollusca, Annelida, and Nematoda, two IF prototypes emerge. The L-type, which includes 35 sequences from 11 protostomic phyla, shares with the nuclear lamins the long version of the coil 1b subdomain and, in most cases, a homology segment of some 120 residues in the carboxyterminal tail domain. The S-type, which includes all four subfamilies (types I to IV) of vertebrate IF proteins, lacks 42 residues in the coil 1b subdomain and the carboxyterminal lamin homology segment. Since IF proteins from all three phyla of the chordates have the 42-residue deletion, this deletion arose in a progenitor prior to the divergence of the chordates into the urochordate, cephalochordate, and vertebrate lineages, possibly already at the origin of the deuterostomic branch. Four phyla recently placed into the protostomia on grounds of their 18S rDNA sequences (Brachiopoda, Nemertea, Phoronida, and Platyhelminthes) show IF proteins of the L-type and fit by sequence identity criteria into the lophotrochozoic branch of the protostomia. PMID- 9847418 TI - Ancient and recent intron stability in the Artemia hemoglobin gene. AB - Artemia has evolved three distinct hemoglobins formed by the association of two nine-domain globin polymers. Sequence analysis of cDNA clones corresponding to two polymers, named T and C, indicates that their genes are the products of a duplication event some 60 million years ago. The present study indicates the presence of 22 introns in each of the T and C polymer genes. The 22 introns are classified into two groups: 17 correspond to positions within globin domains, and 5 correspond to interdomain linkers (or N- and C-terminal extensions). Intron position and reading frame phase are precisely conserved between T and C polymers for all 22 introns, but within each gene the position and phase are not always consistent from domain to domain or from linker to linker. The discordance of Artemia hemoglobin introns is discussed in terms of different model mechanisms and constraints: intron sliding, intron loss or gain, and the exon definition model of primary transcript RNA splicing. The results suggest that constraints of pre-mRNA processing should be considered when considering intron positional changes in homologous genes. PMID- 9847420 TI - Four puzzles about life. AB - To surmount the notorious difficulties of defining life, we should evaluate theories of life not by whether they provide necessary and sufficient conditions for our current preconceptions about life but by how well they explain living phenomena and how satisfactorily they resolve puzzles about life. On these grounds, the theory of life as supple adaptation gets support from the natural and compelling way it resolves the following four puzzles: (a) How are different forms of life at different levels of the vital hierarchy related? (b) Is there a continuum between life and nonlife? (c) Does life essentially concern a living entity's material composition or its form? (d) Are life and mind intrinsically connected? PMID- 9847419 TI - Phylogenetic relationships within the genus Equus and the evolution of alpha and theta globin genes. AB - Sequences of the alpha1, alpha2 and theta globin genes from six equid species have been determined to investigate relationships within the genus Equus. Analyses using standard phylogenetic methods, or an approach designed to account for the effects of gene conversion between the alpha genes, gave broadly similar results and show that the horses diverged from the zebra/ass ancestor approximately 2.4 million years ago and that the zebra and ass species arose in a rapid radiation approximately 0.9 million years ago. These results from the alpha genes are corroborated by theta gene data and are in contrast to mitochondrial DNA studies of the phylogeny of this genus, which suggest a more gradual set of speciation events. PMID- 9847421 TI - The perfect C. elegans project: an initial report. AB - The soil nematode Caenorhabditis Elegans (C. elegans) is the most investigated of all multicellular organisms. Since the proposal to use it as a model organism, a series of research projects have been undertaken, investigating various aspects of this organism. As a result, the complete cell lineage, neural circuitry, and various genes and their functions have been identified. The complete C. elegans DNA sequencing and gene expression mapping for each cell at different times during embryogenesis will be identified in a few years. Given the abundance of collected data, we believe that the time is ripe to introduce synthetic models of C. elegans to further enhance our understanding of the underlying principles of its development and behavior. For this reason, we have started the Perfect C. elegans Project, which aims to produce ultimately a complete synthetic model of C. elegans' cellular structure and function. This article describes the goal, the approach, and the initial results of the project. PMID- 9847422 TI - Phototaxic foraging of the archaepaddler, a hypothetical deep-sea species. AB - An autonomous agent (animat, hypothetical animal), called the (archae) paddler, is simulated in sufficient detail to regard its simulated aquatic locomotion (paddling) as physically possible. The paddler is supposed to be a model of an animal that might exist, although it is perfectly possible to view it as a model of a robot that might be built. The agent is assumed to navigate in a simulated deep-sea environment, where it forages for autoluminescent prey. It uses a biologically inspired phototaxic foraging strategy, while paddling in a layer just above the bottom. The advantage of this living space is that the navigation problem--and hence our model--is essentially two-dimensional. Moreover, the deep sea environment is physically simple (and hence easy to simulate): no significant currents, constant temperature, completely dark. A foraging performance metric is developed that circumvents the necessity to solve the traveling salesman problem. A parametric simulation study then quantifies the influence of habitat factors, such as the density of prey, and body geometry (e.g., placement, direction and directional selectivity of the eyes) on foraging success. Adequate performance proves to require a specific body geometry adapted to the habitat characteristics. In general, performance degrades gracefully for modest changes of the geometric and habitat parameters, indicating that we work in a stable region of "design space." The parameters have to strike a compromise between, on the one hand, to "see" as many targets at the same time as possible. One important conclusion is that simple reflex-based navigation can be surprisingly efficient. Additionally, performance in a global task (foraging) depends strongly on local parameters such as visual direction tuning, position of the eyes and paddles, and so forth. Behavior and habitat "mold" the body, and the body geometry strongly influences performance. The resulting platform enables further testing of foraging strategies or vision and locomotion theories stemming either from biology or from robotics. PMID- 9847423 TI - Self-adaptation in evolving systems. AB - A theoretical and experimental analysis is made of the effects of self-adaptation in a simple evolving system. Specifically, we consider the effects of coding the mutation and crossover probabilities of a genetic algorithm evolving in certain model fitness landscapes. The resultant genotype-phenotype mapping is degenerate in fitness space, there being no direct selective advantage for one probability versus another. Thus there is a "symmetry" between various genotypes that all correspond to the same phenotype. We show that the action of mutation and crossover lifts this degeneracy, that is, the genetic operators induce a breaking of the genotype-phenotype symmetry, thus leading to a preference for those genotypes that propagate most successfully into future generations. We demonstrate that this induced symmetry breaking allows the system to self-adapt in a time-dependent environment. PMID- 9847424 TI - Self-evolution in a constructive binary string system. AB - We examine the qualitative dynamics of a catalytic self-organizing system of binary strings that is inspired by the chemical information processing metaphor. A string is interpreted in two different ways: either (a) as raw data or (b) as a machine that is able to process another string as data in order to produce a third one. This article focuses on the phenomena of evolution whose appearance is notable because no explicit mutation, recombination, or artificial selection operators are introduced. We call the system self-evolving because every variation is performed by the objects themselves in their machine form. PMID- 9847425 TI - Measuring efficacy and safety of different inhaled corticosteroid preparations. AB - Inhaled corticosteroids are the mainstay of treatment for persistent asthma because of their proven efficacy, which is better than any other class of antiasthma therapy. Concerns about unwanted systemic effects with long-term use has, however, limited their use. Efforts have been made to develop inhaled corticosteroids with less systemic activity for a given clinical effect, thereby improving their therapeutic index. Many different study designs and outcome variables have been used to compare different inhaled corticosteroids. Differences in pharmacologic properties between drugs are most easily and accurately measured and quantified by measures of systemic effects. However, these differences should always be related to differences in clinical effects. It is difficult to draw firm conclusions with respect to the therapeutic index of different inhaled corticosteroids because no direct placebo-controlled, dose response comparisons of clinical effects have been made. Despite this caveat, the available studies suggest that microgram for microgram, when delivered by a pressurized metered-dose inhaler (pMDI), fluticasone propionate (FP) is more effective than beclomethasone dipropionate (BDP), triamcinolone acetonide (TAA), or budesonide; however, the efficacy of budesonide delivered by Turbuhaler is equipotent to that of FP delivered by pMDI or Diskhaler and more effective than that of BDP. When comparative safety is considered, budesonide or TAA delivered by pMDI have less systemic activity than FP delivered by pMDI, whereas BDP and FP delivered by pMDI appear to be equivalent. Also, budesonide delivered by Turbuhaler has less systemic activity than FP delivered by Diskhaler. PMID- 9847427 TI - The debate on S-enantiomers of beta-agonists: tempest in a teapot or gathering storm? PMID- 9847426 TI - Molecular modulation of allergic responses. AB - The pathophysiology of allergic diseases involves an intricate network of molecular and cellular interactions. Elevated levels of serum IgE- and TH2 cytokine-associated eosinophilic inflammation characterize allergic diseases and provide potential targets for immunomodulation. Recent evidence that antigen induced allergic responses can be modulated in rodents by mucosal transfer of TH1 cytokine genes or by immunization with plasmid DNAs encoding the sensitizing antigens suggests promising new prophylactic or therapeutic approaches. Innovative research in mapping the regulatory pathways that typify the atopy network will provide a deeper understanding of the molecular mechanisms underlying these diseases and facilitate the design of more specific and efficacious modulation strategies. PMID- 9847428 TI - The value of acoustic rhinometry in assessing nasal responses to cat exposure. AB - BACKGROUND: Acoustic rhinometry (AR) uses sonar principles to map the anatomy of the nasal cavity and has been used in other studies to assess acute airway responses to allergen exposure. OBJECTIVE: The purpose of this study was to evaluate the utility of AR in assessing acute airway responses to cat allergen exposure by using a well-characterized cat exposure model. METHODS: Thirty subjects with a history of cat-induced rhinitis and a positive skin prick test response to cat allergen underwent an environmental cat challenge. Of these 30 subjects, 10 also had repeat challenges at lower levels of antigen to determine whether there was a dose response. Five subjects with negative skin test responses to cat were recruited as control subjects. During the 1-hour cat exposure, upper and lower respiratory symptoms were scored every 5 minutes, and spirometry and AR were obtained every 15 minutes. RESULTS: Although 29 of 30 subjects had changes in AR measurements, no correlations were detected between upper respiratory symptom scores and any of the changes observed in AR. In comparing the baseline challenges with lower antigen level challenges, upper respiratory symptom scores differed significantly (P = .002), whereas AR responses were nearly identical. Subjects without cat allergy did exhibit less response by AR (P = .05 to .13), but the greatest differences remained in the upper respiratory symptoms scores (P < .0001). CONCLUSION: We conclude that although AR does provide an objective measure of nasal response to allergen exposure, it has significant limitations. These are evidenced by the lack of correlation with symptoms, the inability to measure a dose response, and the changes noted even among the control subjects. PMID- 9847429 TI - Comparison of the efficacy of budesonide and fluticasone propionate aqueous nasal spray for once daily treatment of perennial allergic rhinitis. AB - BACKGROUND: Intranasal corticosteroids, such as budesonide and fluticasone propionate, are widely prescribed in the treatment of perennial allergic rhinitis. Once daily budesonide dry powder and fluticasone propionate aqueous suspension have been found to provide similar efficacy in controlling symptoms of perennial allergic rhinitis. OBJECTIVE: The purpose of this study was to assess the efficacy and safety of treatment with once daily budesonide aqueous nasal spray. METHODS: This study involved a multicenter, blinded, randomized, parallel group, placebo-controlled trial of adults with perrenial allergic rhinitis. Patients (n = 273) recorded daily nasal symptoms for 8 to 14 days (baseline) and 6 weeks (treatment). RESULTS: Budesonide decreased combined symptoms to a significantly greater extent than did fluticasone (P =.03); both treatments significantly decreased mean combined nasal symptoms scores compared with placebo. Of the 3 nasal symptoms assessed (ie, nasal blockage, runny nose, and sneezing), nasal blockage was significantly (P =. 009) more decreased with budesonide compared with fluticasone. Both treatments also significantly improved runny nose and sneezing compared with placebo. Improvement in combined nasal symptom scores of the budesonide-treated group reached statistical significance within 36 hours compared with placebo (P =.01); in those patients treated with fluticasone, significant improvement compared with placebo was first observed within 60 hours. Adverse events were mild and transient. CONCLUSIONS: Once daily budesonide aqueous nasal spray, 256 microgram, was significantly better in controlling the symptoms of perrenial allergic rhinitis than once daily fluticasone propionate, 200 microgram, especially nasal blockage. Both treatments were superior to placebo. Budesonide may have a faster onset of action than fluticasone. PMID- 9847431 TI - Differential immune responses to acute lower respiratory illness in early life and subsequent development of persistent wheezing and asthma. AB - BACKGROUND: Recent epidemiologic evidence suggests that 2 wheezing syndromes coexist in early life: transient wheezing, limited to early childhood, and persistent wheezing, which starts in early childhood and persists beyond that age. OBJECTIVE: Whether the nature of the immune response occurring during acute lower respiratory illnesses (LRIs) in infancy differs between these 2 groups of wheezers has yet to be determined. METHODS: We compared total serum IgE levels and peripheral blood eosinophil counts obtained during the acute phase of the first LRI with those obtained during the convalescent phase or with well-baby samples in persistent (n = 49) and transient early wheezers (n = 88), as well as in children who had only nonwheezing LRIs (n = 43) during the first 3 years of life. RESULTS: Total serum IgE levels were significantly higher (P =.008) during the acute phase compared with the convalescent phase of the LRI in persistent wheezers, a response not observed in transient early wheezers (P =.7). Peripheral blood eosinophil counts were significantly reduced during the acute phase of the LRI (P =.009) in transient early wheezers, a response not observed among persistent wheezers (P =.7). Acute responses in children who had nonwheezing LRIs only were similar to those seen in transient early wheezers. CONCLUSION: Alterations in acute immune response to viral infection may be detected at the time of the first wheezing episode in subjects who will go on to have persistent wheezing symptoms. PMID- 9847430 TI - Effect of acyclovir on bronchoconstriction and urinary leukotriene E4 excretion in aspirin-induced asthma. AB - BACKGROUND: Acyclovir (9-[2-hydroxyethoxymethyl] guanine), an inhibitor of the DNA polymerase of the herpes virus, has been reported to exhibit pharmacologic activity other than antiviral activity, including antiasthmatic effects. OBJECTIVE: This study was designed to investigate the protective effect of acyclovir on airway responsiveness to the sulpyrine provocation test and to investigate whether this protective activity is associated with a reduction in aspirin-induced excretion of urinary leukotriene E4 (u-LTE4 ), a marker of cysteinyl leukotriene (LT) overproduction that participates in the pathogenesis of aspirin-induced asthma. METHODS: We assessed the effects of pretreatment with acyclovir on bronchoconstriction precipitated by inhalation of sulpyrine in 16 adult patients with mild or moderate aspirin-induced asthma; those who were in stable clinical condition and were hyperresponsive to the sulpyrine provocation test were allocated to this study. A double-blind, randomized, cross-over design was used. u-LTE4 was measured by a combined reverse-phase HPLC enzyme immunoassay. RESULTS: Acyclovir protects against aspirin-induced attacks of asthma through mechanisms unrelated to its bronchodilator property but related to the improvement of bronchial hypersensitivity to sulpyrine; protection was nearly complete in all patients (P <.0001). By contrast, after acyclovir, the maximum level of u-LTE4 in patients was significantly lower than that in control subjects (P <. 01). CONCLUSION: Our results suggest that acyclovir is not only an antiviral drug but also an inhibitor of analgesic-induced bronchoconstriction, probably acting by inhibiting the release of cysteinyl LTs. PMID- 9847432 TI - A familial predisposition in bronchial hyperresponsiveness among patients with allergic rhinitis. AB - BACKGROUND: Nonasthmatic subjects with allergic rhinitis often have bronchial hyperresponsiveness (BHR). The mechanisms responsible for BHR in asthma include genetic predisposition and airway inflammation, but the causes of BHR in allergic rhinitis are poorly understood. OBJECTIVE: The aim of this study was to investigate whether there is a familial predisposition in allergic rhinitis associated BHR. METHODS: One hundred fifteen children with allergic rhinitis (probands) and their family members underwent methacholine bronchial challenge and skin prick tests with airborne allergens. The probands were divided into 2 groups: BHR(+) (methacholine PC20 <18 mg/mL determined by the dosimeter method; n = 42) and BHR(-) (n = 73). RESULTS: The overall prevalence of BHR was higher in family members of BHR(+) probands than in those of BHR(-) probands (23.3% [27 of 116] vs 10.5% [21 of 200], P <.01). In mothers, this difference was marked (21.4% vs 8.2%, P <.05); a similar trend was observed in fathers (16.7% vs 6.8%) and siblings (34.4% vs 18.5%), although the differences did not reach significance (.05 < P <.1). The bronchial responsiveness index (BR index), a continuous variable derived from the results of methacholine challenge, was significantly higher among family members of the BHR(+) group than those of the BHR(-) group. Furthermore, even when only family members without BHR were considered, the BR index was significantly higher among those (n = 89) of the BHR(+) group than those (n = 179) of the BHR(-) group. There was no difference in atopic status as assessed by the prevalence of atopy (or atopy index) between family members of the BHR(+) group and the BHR(-) group. CONCLUSION: Our results indicated that there is a significant familial predisposition for BHR among patients with allergic rhinitis. Further studies are needed to elucidate whether genetic factors play a role in allergic rhinitis-associated BHR. PMID- 9847433 TI - Effect of rush immunotherapy on airway inflammation and airway hyperresponsiveness after bronchoprovocation with allergen in asthma. AB - BACKGROUND: Rush immunotherapy (RIT) has been shown to be effective in allergic asthma. OBJECTIVE: We investigated the mechanisms of RIT on the basis of cytokine production by T-cell lines and airway inflammation and responsiveness. METHODS: Subjects were 8 patients with house dust mite-allergic asthma treated with dust mite extract RIT for 6 months and 6 RIT-untreated control patients. IL-5 production by Dermatophagoides farinae -specific T-cell lines, eosinophil percentages, and eosinophil cationic protein (ECP) in induced sputum and airway responsiveness to allergen and histamine were evaluated before and after treatment. Changes in eosinophil percentages and ECP in induced sputum and responsiveness to histamine 24 hours after allergen inhalation were also studied. RESULTS: After 6 months of RIT, percentages of total eosinophils (43. 0% +/- 6.90% to 16.8% +/- 2.48%; P <.01), percentages of EG2(+ ) eosinophils (32.6% +/- 6.39% to 19.7% +/- 4.68%; P <.01) and ECP (362.7 +/- 125.3 ng/mL to 26.2 +/- 5.15 ng/mL; P <.05) decreased in induced sputum, and IL-5 production by T-cell lines decreased (617 +/- 93.2 pg/mL to 200.0 +/- 34.1 pg/mL; P <.01). RIT decreased both early- and late-phase bronchoconstriction (early phase: 33.2% +/- 3. 46% to 25.4% +/- 1.42%; P <.03; late phase: 16.2% +/- 3.52% to 6.2% +/- 1.96%; P <.03) and suppressed increases in the percentages of total (61.8% +/- 4.89% to 42.0% +/ 4.67%; P <.01) and EG2-positive eosinophils (55.54% +/- 7.21% to 36.5% +/- 6.43%; P <.01) and ECP (685.6 +/- 217.0 ng/mL to 85.4 +/- 23.4 ng/mL; P <.05) in induced sputum after allergen inhalation. RIT also decreased airway responsiveness to dust mite (1:303.7 +/- 123.7 wt/vol to 1:65.0 +/- 13.2 wt/vol; P <.03) and to histamine before (397.1 +/- 206.9 microgra/mL to 1391.3 +/- 283.3 microgram/mL; P <.03) and after allergen inhalation (139.2 +/- 36.5 microgram/mL to 629.1 +/- 196.3 microgram/mL; P <.03). CONCLUSION: RIT decreases airway inflammation and airway hyperresponsiveness before and after bronchial provocation with allergen, possibly by inhibiting both allergen-specific T-cell- and mast cell-dependent pathways. RIT is an effective antiinflammatory treatment in allergic asthma. PMID- 9847434 TI - Zafirlukast improves asthma symptoms and quality of life in patients with moderate reversible airflow obstruction. AB - BACKGROUND: Previous trials demonstrated the effectiveness of the leukotriene receptor antagonist zafirlukast in patients with mild-to-moderate asthma. OBJECTIVES: We sought to assess the efficacy and safety of zafirlukast and its effect on patients' quality of life (QOL) during a 13-week, double-blind, placebo controlled, multicenter trial in adults and adolescents with moderate reversible airflow obstruction. METHODS: Patients (age range, 12 to 68 years) with total daytime asthma symptoms scores of 10 or greater over 7 consecutive days (maximum, 21/wk), FEV1 45% or greater but less than or equal to 80% of predicted value (>/=6 hours after beta2 -agonist), and reversible airway disease were randomized to 20 mg zafirlukast twice daily (nZ = 231) or placebo twice daily (nP = 223). Efficacy was assessed from changes in daytime and nocturnal symptoms, beta2 agonist use, nasal congestion score, and pulmonary function. QOL was evaluated with a disease-specific Asthma Quality of Life Questionnaire. Safety was determined from adverse event information and clinical laboratory test results. RESULTS: Zafirlukast was significantly (P <.001) more effective than placebo, with reductions from baseline in the daytime asthma symptoms score (-23%), nighttime awakenings with asthma (-19%), and beta2 -agonist use (-24%) and improvements from baseline in morning (+25 L/min) and evening (+18 L/min) peak expiratory flow rates. Compared with placebo, zafirlukast significantly (P /=0.5-unit change from baseline; P N2 disease. The schedule employed was 1.5 Gy 3 times a day in 36 fractions on 12 treatment days without stopping for weekends. The spinal dose has been kept at 40.5 Gy and interfraction interval has been strictly kept at 6 hrs. Clinical, as well as radiological assessment has been encouraging with 23/29 (79.13%) of patients achieving complete regression of the primary tumor and nodal disease. Compared to the conventionally treated controls this value has been significant (P value < 0.001). In stage III disease tumor control has been in 9/10 (90%) cases, while in stage IV disease in 14/19 (73.68%) cases. The acute mucosal reactions have been found to be severe in these cases with 18/29 i.e. 62.06% suffering from grade III reactions. Nasogastric feeding was required in 25.92% of patients. PMID- 9847467 TI - Correlation of radiological and pathological assessment of tumour volume with lymph node involvement in cancer cervix. AB - The present study was carried out in 18 patients with carcinoma cervix stage IB through early IIB-Preoperative computerised tomography (C.T.) was done within seven days before surgery. Volume of tumour was determined from the C. T. films. Radical hysterectomy Type-III was performed in all 18 patients and specimens of cervix, parametrium and lymph nodes subjected to histopathological examination. Each specimen of cervix was cut into 4 to 12 equal sections depending on the size of the tumour mass. Cervical tumour volume was measured. Correlation of radiological with pathological tumour volume and of tumour volume with lymph node involvement was done. C. T. Scan was able to detect tumour mass accurately only four patients (Sensitivity 40%, Specificity 28.5%). In the Indian setting the conventional surgical approach appears to be the more appropriate. PMID- 9847469 TI - Paneth cell metaplasia in colonic adenocarcinoms. AB - The junctional mucosa of 49 colorectal carcinomas were studied for Paneth cell metaplasia. Twenty two cases (44.89%) showed Paneth cells in the junctional mucosa. The carcinomas were seen mainly in the age range of 51 to 60 years (38.4%) The occurrence of Paneth cell metaplasia was higher in the vicinity of tumours of ascending colon and in well differentiated grades of adenocarcinomas. This probably indicates the reactive host response in the junctional mucosa which is of better prognostic significance. PMID- 9847468 TI - Prognostic factors for spinal cord myelitis--an analysis of compiled literature data. AB - The effectiveness of radiotherapy in the treatment of malignant tumors in the head and neck, thorasic and upper abdominal regions is frequently limited by the tolerance of the spinal cord. Therefore knowledge of the factors that influence the tolerance of the spinal cord to radiation is of the upmost important. Safe limits for irradiation of the spinal cord determined in a number of clinical studies is on the conservative side from the point of view of tumor control probability. The bioeffect of a physical dose takes into account the treatment variables and the radiobiological characteristics of the relevant tissue; hence deciding spinal cord tolerance on the basis of bioeffect models would be better approach. The aim of the present study was to analyze the relationship of the prognostic factors with percentage incidence of spinal cord myelitis and to arrive at a spinal cord tolerance bioeffect dose TDF and ERD for optimum incidence of radiation myelitis. PMID- 9847470 TI - Bilateral Wilms tumor: an eleven year experience. AB - Over the last 11 years, 101 patients of Wilms tumor were treated at our hospital. Of these, 7 had bilateral Wilms tumors (BWT). The first two patients were referred in terminal stages and died. Of the latter five, three have survived while one is on treatment. One patient succumbed to liver metastasis, he had stage III disease on one side and was irregular in his chemotherapy protocol. Remarkably, one of the survivors was cured even after developing vertebral metastasis, retroperitoneal recurrence and bilateral lung metastasis. Part of the success was due to "salvage chemotherapy regimens" using Cis-plantinum, ifosphamide and VP-16. Flow cytometric analysis of DNA ploidy performed on only three of the five patients showed that two tumors were aneuploid and one was diploid. Spectacular cure rates can be achieved in BWT by renal conservation surgeries and effective chemotherapy. The use of "salvage chemotherapeutic regimes" and the study of DNA ploidy is being highlighted. PMID- 9847471 TI - A rational approach to alternative systems of medicines. PMID- 9847472 TI - Darkfield microscopic (DFM) and serologic evidences for leptospiral infection in panuveitis cases. AB - 186 out of 226 (82%) panuveitis cases showed the presence of leptospira in their blood samples by dark field microscopy. 75% cases were found positive for leptospira after low speed centrifugation and an additional 7% became positive after high speed centrifugation. Leptospirosis was four times more common in males than in females. The disease was more prevalent in the age group of 15 to 54 years. MAT was performed in 23 cases of which 9 were positive. ELISA was performed in 20 cases of which 9 were positive. DFM was positive in 19 out of these 23 cases. MAT, ELISA and DFM were positive in six cases. Highest antibody titre was found due to L. autumalis alone in two cases, L. autumnalis, and L. pomona in one case, L. bharathy in one case, L. lanka alone in one case and L. pomona one in one case. DFM was found to be more sensitive in a smal number of cases and hence DFM needs further evaluation by other workers in this field. PMID- 9847473 TI - Infantile and childhood types of atopic dermatitis--a clinical comparison. AB - Atopic Dermatitis also known as infantile eczema in infants, is a chronic endogenous eczema which manifests with different characteristic distribution in infants, children and adults. Absence of any reliable laboratory investigation to confirm the diagnosis makes it very difficult to differentiate it from other eczemas especially in infantile and childhood types. This study compares a small group to patients belonging to infantile and childhood types of atopic dermatitis and documents the changes observed in these category of patients. PMID- 9847474 TI - AIDS: knowledge and attitudes of university students in Nagpur. AB - The present cross section-I study was conducted on university students to assess the basic level of knowledge regarding AIDS. Mean score obtained by the Science students related to various aspects of AIDS was significantly higher as compared to Arts and Commerce students 94.4% Science students and 86.7%. Arts and Commerce students expressed their desire to learn more about AIDS. This study has identified considerable extent of knowledge and positive attitudes amongst Science students, but Arts and Commerce students had indequate and poor knowledge. The results of this study show that there is a need of effective health education campaign for this high risk group of individuals which will help them to develop the skills to act on their knowledge and communicate it to others. PMID- 9847476 TI - Paracondylar foramen--a new anomalous aperture of the occipital bone. AB - A new anomalous apperture has been identified by the author in association with the occipital bone. It was situated in the interval between the jugular fossa and the occipital condyles. Out of 118 skull bases examined four instances of this anomalous apperture were found (2 complete and 2 incomplete). The author speculates that they may transmitting the posterior condylar emissary vein in the absence of the condylar canal or in its abrrent course to the jugular foramen. In view of its topographical location the author proposes the name--'Paracondylar foramen' to this anomalous apperture. PMID- 9847475 TI - "Changing trends (clinico-biochemical) in gall-bladder stone disease"--an observation. AB - Present study has been undertaken to know the causative factors responsible for change in trend of gall-stone disease from middle aged, fertile, fat females to young asthenic females in twenties. Our findings reveal high incidence of gall stone formation in non-obese young females. Average fat consumption in non-obese patients was less (17%) than that of obese (26%). However, use of oral contraceptives was high in non-obese females and maximum users were in young age group while in obese in middle age group. Bilirubin content in gall bladder stones of non-obese was significantly more than that of obese (p < 0.01) whereas cholesterol content in gall bladder stones of obese was significantly high when compared to non-obese subjects. Analysis of bile showed significant increase in bilirubin and calcium level of non-obese when compared to control and obese subjects whereas phosphorus levels were significantly decreased in the bile of non obese subjects. These findings suggest that in non-obese females less intake of fat, early use of oral contraceptives, higher contents of bilirubin and calcium and low content of phosphorus in bile may be responsible for gall stone formation. PMID- 9847477 TI - Diabetic blindness in India: the emerging scenario. PMID- 9847479 TI - Prevalence of primary glaucoma in an urban south Indian population. AB - Glaucoma is fast emerging as a major cause of blindness in India. In order to estimate the prevalence of primary open angle glaucoma (POAG) and primary angle closure glaucoma (PACG) in an urban South Indian population, we examined 972 individuals aged 30-60 years, chosen using a cluster sampling technique from 12 census blocks of Vellore town. They underwent a complete ocular examination, including applanation tonometry and gonioscopy, at the Medical College Hospital. Characteristic field defects on automated perimetry was a diagnostic requisite for POAG. Prevalence (95% CI) of POAG, PACG, and ocular hypertension were 4.1 (0.08-8.1), 43.2 (30.14-56.3), and 30.8 (19.8-41.9) per 1,000, respectively. All the PACG cases detected were of the chronic type. Hitherto unavailable community based information on primary glaucoma in our study population indicates that PACG is about five times as common as POAG. PMID- 9847478 TI - Choroidal neovascular membrane. AB - Choroidal neovascular membrane in the macular area is one of the leading causes of severe visual loss. Usually a manifestation in elderly population, it is often associated with age-related macular degeneration. The current mainstay of management is early diagnosis, usually by fundus examination, aided by angiography and photocoagulation in selected cases. Various other modalities of treatment including surgery are being considered as alternate options, but with limited success. The purpose of this review is to briefly outline the current concepts and the management strategy from a clinician's viewpoint. PMID- 9847480 TI - A new curved vitreous cutter for managing phakic retinal detachment with proliferative vitreoretinopathy. AB - In the presence of proliferations anteriorly, adequate excision of the vitreous base is essential. To enable a good vitreous base excision, removal of lens often becomes necessary as it may be damaged while attempting to remove peripheral vitreous. To avoid damage or the need to remove the crystalline lens we have used a new modified curved vitreous cutter along with a wide angle observation system binocular indirect ophthalmomicroscope (BIOM). Use of BIOM during vitreous surgery enables easy viewing of the retinal periphery without the need for scleral depression. Sclerotomies are made as for any regular three-port vitrectomy procedure and the vitrectomy is carried out using the curved vitreous cutter, including the vitreous base, avoiding damage to the crystalline lens. The modified curved vitreous cutter is helpful in removing the peripheral vitreous without damaging the crystalline lens, giving the patient the advantage of intraocular lens implantation at a later date. PMID- 9847481 TI - Triple procedure in posterior segment intraocular foreign body. AB - Three patients with intraocular foreign bodies and traumatic cataracts underwent single stage pars plana lensectomy with anterior capsule preservation, vitrectomy, removal of the foreign body, and intraocular lens implantation. The preserved anterior capsule permitted support for the placement of an intraocular lens in the posterior chamber in the ciliary sulcus. The procedure enabled early visual rehabilitation. This procedure seems useful in the management of posterior segment intraocular foreign body associated with cataract. PMID- 9847482 TI - A method of scoring automated visual fields to determine field constriction causing blindness. AB - Blindness is usually defined by visual acuity criteria. Patients with markedly constricted visual fields are visually impaired even if they have good visual acuity. To our knowledge, no standardised criteria exist to determine the extent of constriction for fields done with the currently used automated static perimetry. The purpose of this study was to suggest a simple method to do so which would help in determining blindness due to field constriction. We reviewed a number of constricted visual fields obtained with Humphrey automated static perimetry. The central 30 degrees field was divided into six concentric zones. By trial and error, we devised criteria for defining visual field constriction based on absolute loss of sensitivity (< or = 0 dB) and relative loss of sensitivity (< or = 5 dB). We suggest that if a zone has at least 75% test points < or = 0 dB and no point > 10 dB, it be considered to have absolute loss of sensitivity for the purpose of defining visual field blindness. Two exceptions to this are also suggested to prevent this criterion from becoming too rigid. Examples are shown to demonstrate application of these criteria in defining blindness due to visual field constriction to < 10 degrees as suggested by the World Health Organization. Standardised determination of visual field constriction with automated perimetry could be useful in more accurate estimation of blindness in surveys, as well as in assessing eligibility for being classified as blind for legal benefits. PMID- 9847483 TI - Microbiological assay of ampicillin in serum and aqueous humor of patients given ampicillin-sulbactam injection. AB - The aim of this study was to determine the bacterial growth inhibitory activities of ampicillin in aqueous humor and serum of patients administered ampicillin sulbactam combination intramuscularly prior to cataract surgery. 43 patients received a combination of both antibiotics intramuscularly at varying periods (60 140 minutes) prior to surgery. Aqueous humor and venous blood were collected at the beginning of the surgery. For microbiological assay, spores of Bacillus subtilis were incorporated in the agar. The test sample and the standard solutions (calibrators) of ampicillin and ampicillin-sulbactam combination were placed in 3 mm wells in the agar. The diameter zones of growth inhibitory activities of ampicillin of the calibrators and the test samples measured in mm were extrapolated to the standard curve and were recorded as ampicillin activity in micrograms/ml. The results of the assay were placed in 5 groups according to the time intervals between injection and collection of serum and aqueous humor (< or = 70, 75, 80, 90, > 90 minutes). Ampicillin activities in sera and aqueous humor of group 5 (> 90 minutes) were significantly higher than the others (p < 0.001). The ratio of ampicillin activities of sera and aqueous humor in group 5 patients was significantly lower indicating higher concentration of ampicillin activity in aqueous humor during this period. Bacterial growth inhibitory activities of ampicillin-sulbactam combination were adequate in aqueous humor of all patients with highest activity being 90 minutes after intramuscular administration indicating the potential usefulness of this antibiotic combination as chemoprophylaxis prior to cataract surgery. PMID- 9847484 TI - Prothrombin time in retinitis pigmentosa. AB - The prothrombin time was recorded for 87 primary retinitis pigmentosa (RP) patients belonging to three different clinical categories. All categories showed prothrombin time higher than normal. There was no correlation between the age of onset and the prothrombin time, nor between duration of disease and the prothrombin time. The high prothrombin time in patients with RP suggests that further study of prothrombin time and related factors may help in better understanding of the pathogenesis of RP. PMID- 9847485 TI - Scientific thinking in ophthalmology. AB - Science, medicine and ophthalmology have all evolved and progressed through varied but powerful influences over the centuries. While the tremendous technological advances in ophthalmology in the past 20 years are readily appreciated, many clinicians fail to grasp the impact of the several clinical trials that have contributed to better patient care. This article briefly traces the history of science, medicine and ophthalmology, and explains how scientific thinking could be applied to the clinical and academic aspects of ophthalmology. PMID- 9847486 TI - Lens expulsion into sub-conjunctival space following peribulbar anaesthesia. PMID- 9847487 TI - Bilateral accessory iris membrane. PMID- 9847488 TI - Sterilisation of tonometers and gonioscopes. AB - Precautions to prevent spread of infection through tonometers and gonioscopes are described in this article. Tonometers and gonioscopes should not be used in the presence of clinically manifest conjunctivitis and keratitis. The Schiotz tonometer should be dipped in a 1:1000 merthiolate solution, and rinsed in saline/distilled water prior to use. The Goldmann applanation prism tip can be wiped with gauze soaked in 70% isopropyl alcohol and then dried before use. Gonioscopes should be cleaned in running water, wiped with gauze soaked in 70% isopropyl alcohol, and then dried before use. Koeppes and goniotomy lenses can be sterilized with ethylene oxide, prior to use in surgery. PMID- 9847489 TI - Planning to reduce childhood blindness in India. AB - Reduction of blindness in children assumes particular significance since a blind child suffers from more blind-years than a blind adult. Estimates of the number of children blind in India and the causes of blindness are relatively crude as there are little reliable epidemiologic data. As a result of this, there is no organised approach to the control of childhood blindness in India. In order to address this issue, a workshop on childhood blindness was held at the L.V. Prasad Eye Institute, Hyderabad in November 1996. The aim of this workshop was to review available data, consider possible strategies, and make recommendations concerning the control of childhood blindness in India. These recommendations along with background information about childhood blindness are presented in this paper. PMID- 9847490 TI - [XXVI Congress of The French Society for Neuroradiology. Paris, France, 30-31 March 1998. Abstracts]. PMID- 9847491 TI - Racial disparity in medications prescribed. PMID- 9847492 TI - Intranasal abuse of prescribed methylphenidate. PMID- 9847493 TI - Atypical antipsychotics. PMID- 9847494 TI - Obsessions associated with hormone therapy. PMID- 9847495 TI - Light therapy for Kleine-Levin syndrome. PMID- 9847496 TI - Psychiatric disorders with onset in the preschool years: I. Stability of diagnoses. AB - OBJECTIVE: To examine the stability of psychiatric disorders with onset in preschool years. METHOD: Five hundred ten children aged 2 through 5 years enrolled initially, with 344 participating in a third wave of data collection 42 through 48 months later. The test batteries used for diagnoses varied by child's age, but they included the Child Behavior Checklist, developmental evaluation, Rochester Adaptive Behavior Inventory and a play session (under age 7 years), and a structured interview (Diagnostic Interview for Children and Adolescents, for parent and child) (ages 7 and older). Consensus DSM-III-R diagnoses were assigned using best-estimate procedures. RESULTS: Intraclass correlations were 0.497 for emotional disorders, 0.718 for disruptive disorders, 0.457 for other diagnoses, and 0.544 for disruptive disorders comorbid with another disorder, indicating moderate stability for all groups of disorders. More than 50% of the children who were aged 2 through 3 years at wave 1 continued to have some psychiatric disorder at wave 2 or 3. Rates were higher for children aged 4 through 5 initially; approximately two thirds were cases subsequently. Odds ratios indicate that having an emotional or disruptive disorder is a strong risk factor for later diagnoses. CONCLUSIONS: While some preschool children in primary care "grow out of" their disorder, an equally large number do not; this finding supports the need for early detection and intervention. PMID- 9847497 TI - Psychiatric disorders with onset in the preschool years: II. Correlates and predictors of stable case status. AB - OBJECTIVE: To examine the correlates and predictors of stability and change in psychiatric disorder occurring among preschool children in a nonpsychiatric, primary care pediatric sample. METHOD: Five hundred ten children aged 2 through 5 years were enrolled; 344 participated in a third wave of data collection 42 through 48 months later. Consensus diagnoses were assigned using best-estimate procedures; variables of maternal psychopathology, family climate, and life stresses were the correlates/predictors studied. RESULTS: For children who were cases initially, family cohesion predicted diagnostic stability. Among initial noncases, those remaining noncases experienced increased family cohesion; for those who later became cases, family cohesion declined. Negative life events declined when children were consistently noncases. Children who were initially noncases but were cases at the two subsequent waves had the highest levels of maternal negative affect. Predictors at wave 1 for wave 2 cases status included lower socioeconomic status, less family cohesiveness, and greater family inhibition/control. Wave 2 correlates of wave 2 status included older children and negative life events. Wave 2 predictors of wave 3 status included being older, while wave 3 correlates of wave 3 case status included older children and higher maternal negative affect. CONCLUSIONS: Family context contributes to the maintenance and onset of problems beginning in the preschool years. PMID- 9847498 TI - Clinical and treatment characteristics of children with attention deficit/hyperactivity disorder in psychiatric practice. AB - OBJECTIVE: To capture information about the clinical characteristics of, and treatments for, children with attention-deficit/hyperactivity disorder (ADHD) in psychiatric practice. METHOD: A mailed, self-administered questionnaire was sent to 81 practicing psychiatrists for them to complete on the next three consecutive patients aged 14 years and younger with ADHD seen during the 12-day study period. Information collected included the sociodemographic, clinical, and treatment characteristics of sampled patients. RESULTS: Patients in the study were predominantly white (85%), male (78%), and between 10 and 14 years old (58%). The most common ADHD subtype was combined/predominantly hyperactive (86%); 31% had no other comorbidity. Ninety-seven percent were receiving medications, with 49% receiving two or more. The single most common medication reported was methylphenidate (51% of patients) followed by clonidine (20%). Psychotropics other than psychostimulants were used in a majority of patients (55%). CONCLUSIONS: Psychiatrists, and child and adolescent psychiatrists in particular, see a more severely impaired and complex group of patients than would be expected of primary care providers. The treatment patterns of psychiatrists for these patients do not reflect the simpler treatments usually studied in clinical trials. PMID- 9847499 TI - An open trial of bupropion for ADHD in adolescents with substance use disorders and conduct disorder. AB - OBJECTIVE: Adolescents with conduct disorder (CD) and substance use disorders (SUD) have higher rates of comorbid attention-deficit/hyperactivity disorder (ADHD) than those without CD and SUD. Comorbid ADHD may contribute to more severe SUD. Treatment of ADHD may enhance effective treatment of substance abuse and behavior problems, yet there are few data regarding pharmacological treatment of ADHD in such youths. METHOD: Pilot data are presented from a 5-week, open-label bupropian trial in 13 nondepressed adolescent boys in a residential treatment program focusing on substance and behavioral treatments. All had diagnoses of ADHD, CD, and SUD. Patients' doses were titrated to a maximum fixed daily dose of 300 mg of bupropion. The Conners Hyperactivity Index and Daydream Attention scores, along with Clinical Global Impressions Severity of Illness (CGI-S) ratings, were obtained at baseline and at the fifth week of treatment. RESULTS: Subjects' mean Conners Hyperactivity Index score declined from 75.5 to 65.4 (p < .01, Wilcoxon signed rank test) (13% decline). The mean Daydream Attention score declined from 59.5 to 53.6 (p < .02, Wilcoxon signed rank test) (10% decline). Mean CGI-S declined from 4.9 to 3.0 (p < .002, Wilcoxon signed rank test) (39% decline). CONCLUSIONS: These preliminary data suggest that bupropion may be a useful treatment for ADHD in adolescents with CD and SUD; they suggest the need for a controlled trial of bupropion in such youths. PMID- 9847500 TI - Death concepts in suicidal adolescents. AB - OBJECTIVE: To investigate the relationship between components of death concept (preoccupation with death, death as a pleasant state, and death as final) and suicidal behavior in adolescents. METHOD: The death concepts of 51 suicidal inpatients, 102 nonsuicidal inpatients, 36 emergency room suicidal subjects, and 81 normal controls were compared using Pfeffer's Child Suicide Potential Scale. In addition, the IQ level as well as emotions that potentially influence the death concept were measured. RESULTS: Both groups of suicidal adolescents evaluated death as more pleasant than the nonsuicidal groups. All the study groups equally perceived death as a final state. Suicidal inpatients were more preoccupied with death than nonsuicidal inpatients, but surprisingly among all study groups, including normal controls, the emergency room suicidal subjects were the least preoccupied with death. Partialing out depression, anxiety, and aggression specifically augmented the association between preoccupation with death and suicidality. Thus the relationship between death concept and suicidality appears to be a direct one. No correlation was found between suicidality and intelligence level. CONCLUSIONS: Elements of death concept distinguish suicidal from nonsuicidal as well as between hospitalized versus nonhospitalized suicidal adolescents. Thus the death concept evaluation is potentially valuable in the assessment of adolescents with a high risk for suicide. PMID- 9847501 TI - Reasons for adolescent suicide attempts: associations with psychological functioning. AB - OBJECTIVE: To determine reasons for suicide attempts in adolescents and to examine the relationship between these reasons and psychological functioning. METHOD: Self-reported reasons for suicide attempts and psychological functioning were examined in 120 adolescent suicide attempters who presented to a pediatric general hospital. RESULTS: Consistent with prior research, the most frequently endorsed motives for self-harm were to die, to escape, and to obtain relief. More manipulative reasons for overdose (such as making people sorry) were endorsed less frequently. Adolescents who cited death as a reason for their suicide attempt reported more hopelessness, socially prescribed perfectionism, depression, and anger expression. Discriminant function analyses indicated that high levels of depression and anger expression predicted a self-reported wish to die, and high levels of depression and socially prescribed perfectionism predicted death as the primary reason reported for the suicide attempt. CONCLUSIONS: Systematic assessment of the reasons for a suicide attempt is a useful tool for clinicians in determining recommendations for follow-up treatment. PMID- 9847502 TI - Suicidal thinking among adolescents with a history of attempted suicide. AB - OBJECTIVE: To examine the risk of suicidal plans and ideation, depression, and other factors (low self-esteem, loneliness, fatalism, pessimism) among adolescents with a lifetime history of attempted suicide. METHOD: A self administered questionnaire was used in a school-based survey of five middle schools (grades 6 through 8) enrolling 6,400 students. Usable questionnaires were obtained from 5,423 (85.3%). Data were obtained on a wide range of constructs including recent and lifetime suicide attempts, recent and lifetime suicide plans, recent ideation, symptoms of DSM-IV major depression, self-esteem, pessimism, loneliness, and fatalism. RESULTS: Data on crude prevalence showed thoughts about death, wishing to be dead, thoughts of suicide, and suicide plans were all significantly higher among youths with a history of attempts. Suicidal thinking was related to being more lonely, more fatalistic, and more pessimistic, and to less self-esteem, in addition to depression and a history of attempts. Multivariate analyses revealed the strongest factors associated with current suicidal thinking were history of attempts (odds ratio [OR] = 3.50), depression (OR = 5.34), and recent life stress (OR = 2.64). Compared with youths with none of the factors examined, those with six or more were at extreme risk (OR = 67.87). CONCLUSIONS: The strong association between history of suicide attempts, current ideation, and depression indicates that past suicide attempts occur in the context of other signs of psychosocial dysfunction. Given the paucity of epidemiological data on the natural history of suicidal behaviors among youths, more epidemiological studies of the antecedents and consequences of the range of suicidal behaviors among children and adolescents are needed. Given the high risk of subsequent suicidal behaviors by youths who have attempted but not completed suicide, this constitutes a high-risk population on which future research should focus. PMID- 9847503 TI - Attitudes toward psychiatric medications among incarcerated female adolescents. AB - OBJECTIVE: Beliefs about and attitudes toward pharmacotherapy have been shown to play an important role in mediating compliance with psychoactive medications. This study is an examination of attitudes toward drug treatment in a sample of delinquent incarcerated girls. METHOD: Female adolescents (N = 214) in a juvenile detention facility were administered a questionnaire assessing general beliefs and specific concerns about taking psychiatric medications. Demographic and historical data, including prior psychiatric drug treatment and illicit substance abuse, were also examined as possible influencing variables. RESULTS: Nearly one half of subjects were skeptical about the benefits of pharmacotherapy. Prior illicit drug use did not influence attitudes, but having been treated with psychiatric medication was related to enhanced perceptions of the efficacy and acceptance of such therapy. Specific concerns about untoward effects of pharmacotherapy were identified. CONCLUSIONS: Results suggest that these delinquent adolescents would benefit from techniques to improve initial attitudes toward treatment and challenge the prescribing physician to consider the unique worries and concerns that may undermine compliance with medication regimens among teenagers. PMID- 9847504 TI - Linking self-reported childhood behavioral inhibition to adolescent social phobia. AB - OBJECTIVE: Behavioral inhibition in children has been hypothesized to be a risk factor for the later development of social phobia. However, this hypothesis has yet to be demonstrated in a prospective study. The purpose of the study presented here is to test whether behavioral inhibition in childhood constitutes a risk factor for social phobia during adolescence. METHOD: The sample consisted of 2,242 high school students assessed over a 4-year period. Assessments included self-report questionnaires, structured clinical interviews, and measurements of heart rate. Cox proportional hazards models were used to evaluate risk. RESULTS: Social avoidance, a component of behavioral inhibition, predicted onset of social phobia during high school. However, social avoidance was not related to depression in adolescence. Another component of behavioral inhibition, fearfulness, increased the risk for both social phobia and depression. Among subjects who were both socially avoidant and fearful, 22.3% developed social phobia--a risk more than four times greater than that for subjects with neither feature of behavioral inhibition. CONCLUSION: This prospective study demonstrates that behavioral inhibition in childhood increases the risk of social phobia in adolescence. PMID- 9847505 TI - Effect of sibling perception of differential parental treatment in sibling dyads with one disabled child. AB - OBJECTIVE: To examine sibling perception of parental differential treatment in families of children with pervasive developmental disorder (PDD), Down syndrome (DS), and nondisabled controls. METHOD: Sibling self-concept and social support were studied in the context of sibling perceptions of parental differential treatment and caretaker plus teacher evaluations of sibling behavioral adjustment. Measures were completed at time 1 and time 2. The effect of parental stress and the difficulty of the disabled child, as well as the sibling relationship, were considered. RESULTS: For siblings of PDD children, internalizing and externalizing behavior problems identified by caretakers were evident at time 1 and more accentuated at time 2, at which time teachers also identified these difficulties. DS siblings were reported by caretakers and teachers to have only internalizing problems and only at time 2. These difficulties related to the perception for PDD siblings that they were preferred over their disabled sibling and for the DS sibling that their disabled sibling was preferred. Feelings of low competence predicted internalizing difficulties. Social support, especially over time, had a positive effect for all siblings, including the controls. CONCLUSION: The elucidation of specific mechanisms contributing to adjustment problems in the siblings of disabled children will allow for the development of specific preventive interventions. PMID- 9847506 TI - Psychopathology in non-clinically referred sexually abused children. AB - OBJECTIVE: To determine the prevalence of psychiatric symptoms and disorders found in non-clinically referred, sexually abused children (nCR-SAC), aged 6 to 16 years, during the 30- to 60-day period after disclosure and termination of abuse. METHOD: Eighty nCR-SAC were compared with clinical and nonclinical groups of nonabused children matched by age, race, and socioeconomic status. Structured and semistructured interviews and standardized rating scales were used for assessment. RESULTS: More posttraumatic stress disorder (PTSD) was found in the nCR-SAC than in the two comparison groups, and symptom severity was intermediate, except for symptoms of posttraumatic stress, trait anxiety, and depression, which were highest in the nCR-SAC. CONCLUSIONS: Sexually abused children are at high risk for PTSD and symptoms of posttraumatic stress, anxiety, and depression in the immediate period after disclosure and termination of abuse. Findings indicate the need for routine and systematic evaluation for these symptoms and PTSD for treatment planning. PMID- 9847507 TI - Factors contributing to the generational transmission of child maltreatment. PMID- 9847508 TI - Development of the cerebral cortex: XII. Stress and brain development: I. PMID- 9847509 TI - [Nuclear oncology with monoclonal antibodies and peptides]. AB - Imaging and therapy using radiolabeled monoclonal antibodies have proved useful in many clinical studies. However, immunogenicity of mouse antibodies to human and insufficient tumor-to-normal tissue ratios remained to be solved. Chimerization and humanization by genetic engineering, and multistep targeting techniques have enabled lower immunogenicity and higher tumor-to-normal tissue contrast. Peptides like somatostatin-analogs have been reportedly useful in imaging tumors, which are either somatostatin receptor positive or negative. Elevated normal tissue accumulation of radiolabeled peptides is a drawback in aiming internal radiation therapy. PMID- 9847510 TI - [Comparison of single sample methods for determination of plasma clearance using 99mTc-MAG3]. AB - We compared relationship among three single blood sampling methods which have been proposed for determination of plasma clearance with 99mTc-MAG3. Russell's and Bubeck's algorithms were employed for 48 adults (average age: 53 years) and Bubeck's and Piepsz's algorithms were used in 9 children (average age: 5.8 years). Three methods showed very high correlation among them. However, Russell's and Piepsz's methods were about 1.4 times higher than Bubeck's method in calculating plasma clearance of 99mTc-MAG3. The present study indicates that three methods discussed here are useful for the determination of plasma clearance with 99mTc-MAG3. In addition, Bubeck's method may be preferable as a reference for single plasma sampling method because of common application to adults and children. PMID- 9847512 TI - [On the application of the triple energy window method to the 201Tl imaging]. AB - In the radionuclide imaging by 201Tl several K-X rays are acquired, so it is difficult to eliminate scattered photons in the acquired photons. This paper describes the limitation and efficiency of the triple energy window (TEW) method in the application of 201Tl myocardial studies. We evaluated the TEW method by simulations and the setting of the parameters in the TEW method were 20, 30 and 40% for the width of the main window, 3, 5 and 7% for the width of the two sub windows. Moreover, the location of the main window was shifted by each 1 keV from 60 to 80 keV. Simulation results were evaluated with the mean square error between true scattered photons and estimated scattered photons. The results show that the mean square error was minimized when the widths of the main window were 20, 30 and 40%, and the centers of the main window were 66-68, 69-70 and 72-73 keV, respectively. PMID- 9847511 TI - [Quantification of cerebral blood flow with 99mTc-ECD SPECT based on a 3 compartment model]. AB - In the present study we developed a method for quantifying regional cerebral blood flow (rCBF) using 99mTc-ECD SPECT based on a 3-compartment model. The dynamic SPECT scanning and sequential sampling of arterial blood were performed on 12 subjects with cerebrovascular diseases and etc. We defined brain fractionation index (BFI) as a parameter of rCBF, which was obtained from a single SPECT data and arterial input. The relationship between the values of BFI and rCBF obtained by the 133Xe inhalation method was analyzed by approximation with exponential function. In this method, rCBF was calculated from the values of BFI using the inverse function of the exponential function as a regression curve. The method was applied seven other patients with cerebrovascular diseases and the values of rCBF were compared with those obtained by the 133Xe inhalation method. We observed a good correlation (r = 0.854), and the inclination was approximately 1. This method can be applied to not only large field SPECT cameras but also conventional SPECT cameras. PMID- 9847513 TI - [Discrepancy between 99mTc-HMPAO and 99mTc-ECD in Alzheimer's disease: does the retention mechanism depend on the disease?]. AB - The discrepancy between 99mTc-hexamethyl-propyleneamine oxime (HMPAO) and 99mTc ethyl cysteinate dimer (ECD) in Alzheimer's disease (AD) was investigated and compared with in cerebral ischemic disease (CID). The subjects were fourteen AD and thirty-one CID patients with clinically reasonable rCBF reduced lesion on 133Xe SPECT. The subjects did not include the cases of acute and subacute CID. These SPECT were performed within two weeks by ring-type dynamic SPECT (HEADTOME, Shimadzu, Japan). In the CID group, both of HMPAO and ECD SPECT could hardly detect the mildly reduced rCBF lesion on 133Xe SPECT but normal on X-CT. In the case of AD group, the rCBF-reduced lesion on 133Xe SPECT could be detected well by ECD SPECT, but the HMPAO hardly detected the reduced lesion. This discrepancy between HMPAO and ECD may be due to the difference of the retention mechanism. In the case of AD, the injury of esterase activity that participates with the ECD retention may be more notable than that of glutathione activity for the HMPAO retention. These results suggest suggest that the reduction of ECD or HMPAO density depends directly on the insufficiency of retention mechanism rather the than rCBF reduction. And the insufficiency of this retention mechanism depends on also type of the disease i.e. AD or CID. PMID- 9847514 TI - [The compensation method for the effects of system resolution to the left ventricular volume measurements using QGS program]. AB - We evaluated the accuracy of left ventricular volume calculations performed using the left ventricular function automatic analysis software (QGS program), which automatically extracts the left ventricular profile from ECG-gated myocardial SPECT data and then measures the left ventricular volume. We used a myocardial phantom for this evaluation. The results of our experiments showed that the left ventricular volume was underestimated by up to 13% as the SPECT spatial resolution deteriorated. To overcome this problem, we have developed a method for correcting the left ventricular volume measured by the QGS program according to the SPECT spatial resolution of the detectors. This new correction method permits the left ventricular volume of the phantom to be calculated with an error of less than 1.4%. PMID- 9847515 TI - [Markedly decreased uptake with 123I-MIBG myocardial scintigraphy in a case with vasospastic angina]. AB - A 68-year-old female whose myocardial sympathetic function was severely damaged with multi-vessel vasospastic angina is presented. She had no signs of autonomic dysfunction or diabetes mellitus. Myocardial imaging with 123I-MIBG showed extremely diminished uptake, but 201TlCl and 123I-BMIPP SPECT images were almost normal. Coronary arteriography revealed no significant atherosclerotic stenosis, multivessel spasm was observed by provocation test using acetylcholine. The extremely diminished uptake of 123I-MIBG was slightly increased in response to medication and the subsequent improvement of the patient's condition. Markedly decreased uptake with 123I-MIBG myocardial scintigraphy was considered to be due to multi-vessel spastic angina. We believe that this method of imaging study is useful for evaluating the healing stage of myocardial sympathetic dysfunction. PMID- 9847516 TI - [Application of a semi-automatic ROI setting system for brain PET images to animal PET studies]. AB - ProASSIST, a semi-automatic ROI (region of interest) setting system for human brain PET images, has been modified for use with the canine brain, and the performance of the obtained system was evaluated by comparing the operational simplicity for ROI setting and the consistency of ROI values obtained with those by a conventional manual procedure. Namely, we created segment maps for the canine brain by making reference to the coronal section atlas of the canine brain by Lim et al., and incorporated them into the ProASSIST system. For the performance test, CBF (cerebral blood flow) and CMRglc (cerebral metabolic rate in glucose) images in dogs with or without focal cerebral ischemia were used. In ProASSIST, brain contours were defined semiautomatically. In the ROI analysis of the test image, manual modification of the contour was necessary in half cases examined (8/16). However, the operation was rather simple so that the operation time per one brain section was significantly shorter than that in the manual operation. The ROI values determined by the system were comparable with those by the manual procedure, confirming the applicability of the system to these animal studies. The use of the system like the present one would also merit the more objective data acquisition for the quantitative ROI analysis, because no manual procedure except for some specifications of the anatomical features is required for ROI setting. PMID- 9847517 TI - [Improved the accuracy of 99mTc-MAG3 plasma clearance method: the problem of the calculated plasma volume and its modification]. AB - The 99mTc-MAG3 plasma clearance method (MPC method), reported by Oriuchi et al., is a simple and useful count-based gamma camera method for calculating the 99mTc MAG3 plasma clearance (CLMAG). However, a discrepancy of CLMAG calculated by MPC method (MPC-CLMAG) from the tubular extraction rate (TER) calculated by Russell's single-sample clearance determination (Russell-TER) was noted. The calculated plasma volume is assumed to be the cause. Since the plasma volume is reported to have a linear correlation with body surface area, Dissmann's formula was applied to calculate the plasma volume. Then Dissmann's formula was replaced by Ogawa's formula in the MPC method, and the procedure was then called the modified MPC method. The CLMAG were obtained using MPC method, modified MPC method and the TER was obtained Russell's method in 95 patients with urological disorders. Then the MPC-CLMAG and modified MPC-CLMAG were compared with Russell-TER. Comparison of the MPC-CLMAG with the Russell-TER demonstrated a coefficient of correlation of 0.82, but dissociation of the slope of regression lines was found between males and females. The modified MPC-CLMAG improved the coefficient of correlation to 0.92, and diminished the dissociation of the slope of regression lines between males and females. We verified that the dissociation was due to the plasma volume calculated by Ogawa's formula. Ogawa's formula included hematocrit, body weight, body height and different coefficients for gender. The plasma volume calculated by Ogawa's formula were lower in males and higher in females than that calculated by Dissmann's formula. And marked discrepancy in the plasma volume in patients with a body surface area below 0.5 m2 was observed. So the MPC method might become more accurate by substituting Dissmann's formula for Ogawa's formula resolting in a method that is applicable to both males and females, children and adults in clinical use. PMID- 9847518 TI - [Evaluation of a new method for susceptibility testing against fastidious and unusual bacteria]. AB - The susceptibilities of 154 clinical isolates of fastidious and unusual bacteria were determined by the Etest, and were compared with the MICs for these strains obtained by the agar dilution method. Antimicrobial agents; benzylpenicillin, ampicillin, cefotaxime, imipenem, erythromycin, clindamycin and minocycline were used in this study. In general, the Etest MICs were rapidly and easily interpreted. The Etest MICs agreed within +/- 1 log2 and +/- 2 log2 dilution, 87.6% and 98.2% respectively in all strains tested. MICs of vancomycin with the Etest for S. pneumoniae and other Streptococci tended to be 1 to 2 log2 dilution higher than those of the agar dilution method. MICs of erythromycin for the other Streptococci also tended to be 1 to 2 log2 dilution higher. However, there were no significant differences for H. influenzae, Campylobacter spp., H. pylori, Capnocytophaga spp., E. corrodens, M. catarrhalis, N. gonorrhoeae, Corynebacterium spp., L. monocytogenes and Anaerobic bacteria. As mentioned above, the Etest represent a potential method for antimicrobial susceptibility testing of these fastidious and unusual bacteria. PMID- 9847519 TI - In vitro cytotoxic effects of vacuolating cytotoxin produced by clinical isolates of Helicobacter pylori. AB - The vacuolating cytotoxin produced by Helicobacter pylori is considered to be one virulence factor causing peptic ulceration. In this study, we examined the activity of vacuolating cytotoxin in induction of intracellular vacuolation of rabbit gastric epithelial cells (RGECs). We used culture supernatants of H. pylori as a source of vacuolating cytotoxin and quantitated cytotoxic activity by the MTT method. Intracellular vacuolation of RGECs was observed in the presence of 36 of 57 (63%) clinically isolated H. pylori strains. However, there were no differences in the incidence of H. pylori strains with positive vacuolating cytotoxin (Tox+) among patients with gastritis, gastric ulcers or duodenal ulcers. The MTT assay showed that the cytotoxic activity of H. pylori supernatants obtained from patients with gastric ulcers was significantly higher than in patients with gastritis (p < 0.01), but was not different to duodenal ulcer patient supernatants. Similar results were also observed in Tox+ isolates, however, there were no significant differences between patients with regard to the incidence of vacuolating cytotoxin-negative isolates. Although our data may not indicate a clear correlation between prevalence of vacuolating cytotoxin and clinical manifestations, they suggest that H. pylori harboring vacuolating cytotoxin may particularly induce damage to the gastric epithelium in patients with gastric ulcers. PMID- 9847520 TI - [Analysis of utility of the phenotyping method on detection of cases infected by multiple strains of methicillin-resistant Staphylococcus aureus (MRSA)]. AB - In this study, we compared the types of methicillin-resistant Staphylococcus aureus (MRSA) isolated from several foci of the same patient to find the incidence of multiple strain infection of MRSA in bacteremia cases. We will also evaluate the utility of the typing methods of phenotyping and genotyping for the above mentioned objective and judge the dissimilarity of clinical characteristics between the single strain infection and multiple strains infection. We studied 21 cases of MRSA bacteremia who were culture-positive both from blood and other foci in the same patient at Nagasaki University Hospital during 1990-1994. Clinical data were retrospectively collected from the patients' records. Phenotyping of all 113 MRSA isolates were done by coagulase typing (I-VIII), production of enterotoxins (SEA-SED) and toxic shock syndrome toxin-1 (TSST-1), hemolysis typing and antibiogram (MIC). In addition, typing of the same isolates were done by Pulsed-Field Gel Electrophoresis (PFGE), using Gene Navigator System as the genotyping. Several types of MRSA were found from different foci in the same patient in 8 of 21 cases (38%) by phenotyping. The same typing results were obtained in 7 of 8 the multiple strains isolated cases by PFGE. Two types were obtained from another case by phenotyping, but by PFGF, 3 types were obtained. We consider that phenotyping method is convenient and reliable for judgment of the difference in types isolated from different foci in the same patient, but PFGE possibly provide us more detailed epidemiological information. The epidemiological investigation must be done very carefully, especially in immunocompromised hosts as MRSA bacteremia cases, because the chance of multiple strains infection is relatively high among these cases. PMID- 9847521 TI - [Evaluation of a PCR assay commercial kit for quantification of human immunodeficiency virus type-1 RNA]. AB - We studied a basic evaluation of the reliability and usefulness of the test results to assess the validity of the kit "AMPLICOR HIV-1 Monitor" as a laboratory tool, by determining the reproducibility, linearity on dilution, possible effects of interference on the results, and correlation with the results obtained at outside facilities. Furthermore, we compared the HIV-1 RNA load between blood samples obtained from HIV-1 subtype B and E. The HIV-1 RNA load measurement was made according to the pre-determined methods of this kit, in blood samples obtained from HIV-positive outpatients. Simultaneous reproducibility was 23.08%-32.95% in C.V.% and linearity was maintained between 110 copies/ml and 2,184,277 copies/ml, demonstrating favorable performance of the kit. The institution correlation between two facilities were also favorable. Fluctuation of measurement by interference was absent for bilirubin, hemoglobin and chyle, but was significant for heparin. PMID- 9847522 TI - [Anti-rabies antibody levels observed in subjects who were bitten by supposed rabid animals abroad and received post-exposure immunization]. AB - From December of 1990 to December of 1997, 119 subjects visited to our hospital to receive post-exposure therapy using purified chick embryo cell rabies vaccine manufactured by the Chem-Sero-Theraptic Institute (Katestuken), because they had been bitten by supposed rabid animals abroad. The forty of the subjects (male: 25, female: 15) wished to have their anti-rabies antibody levels examined. The number of samples taken after 5 or 6 shots rabies vaccine were 30 and 15, respectively. The antibody levels after 6 shots of rabies vaccine varied from 1.0 IU/ml to 10.1 IU/ml. After 5 shots the antibody levels fluctuated from under 0.1 IU/ml to over 8.8 IU/ml, and 3 subjects were found to have antibody titers of under 0.5 IU/ml which is the WHO minimal protective level. Two of these 3 subjects found to have antibodies of 1.0 IU/ml and 3.1 IU/ml. after the 6th injection. However, these 3 subjects had the hazard to have rabies despite post exposure immunization, because the incubation period of rabies is found to be 1-3 months in about 60% of the cases. The potency of Kaketsuken's rabies vaccine should be increased to provide higher antibody levels. PMID- 9847523 TI - [Isolation of Legionella and free-living amoebae at hot spring spas in Kanagawa, Japan]. AB - Microbiological contamination of hot spring bath water is a public health concern. A province-wide survey was carried out to determine the extent and distribution of both Legionella and free-living amoebae contamination. Among 30 samples of hot spring bath from 12 sites in Kanagawa, Japan, L. pneumophila was detected in 21 water samples from 11 sites, ranging from 10(1)-10(3) CFU/100 ml. Serogroups 3, 5 and 6 of L. pneumophila were predominantly isolated from the samples. Naegleria (46.7%), Platyamoeba (33.3%), Acanthamoeba (10.0%) and 2 other genera of free-living amoebae were detected in 22 samples from 11 sites. One or more genera of host amoebae of Legionella occurred in 17 samples (56.7%) from 9 sites. Another thing to be noted is that 13 water samples contained N. lovaniensis. Although N. lovaniensis is nonpathogenic, it is considered an indicator organism for places that are suitable for the growth of N. fowleri, a causative agent of primary amoebic meningoencephalitis in man. PMID- 9847524 TI - [Occurrence of free-living amoebae and Legionella in whirlpool bathes]. AB - Occurrence of both Legionella species and free-living amoebae were surveyed in whirlpool bathes installed in 11 private houses, 8 public bathes and 13 spas. Free-living amoebae that are known to be the hosts of Legionella were isolated from 24 out of 32 water samples (75%). Single Legionella species, L. pneumophila, with different serogroups (SG) predominantly SG3 (18.3%), SG5 (23.7%) and SG6 (15.8%), were isolated from 21 damples, ranging from 10(1) to 10(4) CFU/100 ml. Further studies were conducted for 10 consecutive weeks to monitor the occurrence of both free-living amoebae and Legionella in the whirlpool bathes of 4 private houses. Free-living amoebae, such as Hartmannella and Vexillifera, and L. pneumophila SG1, SG3, SG4, SG5 and SG6 were consistently isolated from all the water samples throughout the monitoring periods. Bath basins in which Hartmennella and Vannella were isolated tended to harbor large number of Legionella. Management practices such as frequent washing filter elements and/or frequent addition of tap water to bath basins is highly recommended to reduce microbial contaminants. PMID- 9847525 TI - [Experimental infection of Naegleria fowleri in mice]. AB - Ten SPF mice (ddY, 4w-old, female) were infected by nasal instillation with an isolate of Naegleria fowleri that was first isolated from a patient with primary amoebic meningoencephalitis (PAM) in Japan. Of these mice, 2 showed clinical signs typical for PAM on the 4th day. On the next day, 5 mice became very ill and remained immobile; their movement and response to painful stimuli diminished progressively. All the infected mice were then examined histopathologically on the same day regardless of their clinical signs. Pathological changes due to invasion and/or proliferation of amoebae were observed in 5 mice with clinical signs. Swelling of the nasal mucosa and ulcerated nasal epithelium with inflammatory cells were observed. Proliferation of amoebae was detected to a lesser extent in nasal cavity including mucous membrane and nasal epithelium. Olfactory lobes and arteriolar hemisphere were necrotic with haemorrhage and filled with amoebae. From these findings the pathogenicity of the isolate was confirmed to develop PAM in experimental animals. It was also observed that the olfactory neuroepithelium was the route of invasion in PAM due to N. fowleri and consequently migration occurred through olfactory lobes into the cerebrum. PMID- 9847526 TI - [Two cases of Achromobacter xylosoxidans sepsis]. AB - Achromobacter xylosoxidans is a gram-negative bacterium whose natural habitat has not been clearly defined. It has been isolated from ear discharge and the large intestine of humans and from various hospital or environmental water sources. Infection with A. xylosoxidans in humans has been documented, and resulting illnesses include meningitis, pneumonia, cholecystitis, peritonitis and urinary tract infection. Bacteremia due to A. xylosoxidans is rare, and little information on treatment is available. Two cases of bacteremia due to A. xylosoxidans in patients with hemapoietic malignancies are reported herein. Case 1 involved a 70-yr. male whose clinical diagnosis was IgA lambda-type plasmacytoma. Case 2 involved 72-yr. male whose clinical diagnosis was acute lymphatic leukemia (L2). Both patients had been catheterized. Neutropenia was noted and the white blood cell counts were 20/microliter in case 1 and 35/microliter in case 2 when A. xylosoxidans was isolated from the blood culture. We suggest that bacteremia due to A. xylosoxidans may have been related to the presence of the catheter and neutropenia. PMID- 9847528 TI - [A case of hypoxemic acute bronchiolitis presenting with diffuse nodular shadows caused by Mycoplasma pneumoniae]. AB - A fifty year-old female who had previously been well presented with a productive cough and a high fever. Her initial chest X-ray film showed no abnormal lung shadows. Despite partial improvement of the fever and the serum level of acute phase reactant (CRP) in response to intravenous administration of piperacillin, she complained of increasing severity of cough and dyspnea. Follow-up chest X-ray films taken five days after therapy with piperacillin showed diffuse nodular shadows in the mid-to-lower lung fields bilaterally. Chest CT scan disclosed diffuse miliary nodules at the lung periphery and thickening of bronchovascular markings. Chest auscultation revealed late inspiratory coarse crackles and expiratory wheezing, and the patient's arterial oxygen tension was 61 mmHg. Suspected of suffering from primary atypical pneumonia, she was started on therapy with intravenous minocyclin (200 mg/day), two days after treatment her symptoms began improving significantly. Anti-mycoplasma antibody was found to be x 1280, and cold hemoagglutinin x 1024, establishing the diagnosis of Mycoplasma pneumoniae infection. The patient's condition completely recovered following a one week treatment with minocyclin. We concluded that her respiratory infection was caused by piperacillin-sensitive mico-organism, and also Mycoplasma pneumoniae which brought about hypoxic acute bronchiolitis to the patient. PMID- 9847527 TI - [Legionella longbeachae pneumonia in a gardener]. AB - A 52-year-old male gardener, who traveled to Guam Island several days ago, was admitted to our hospital with fever, cough and dyspnea. His chest X-ray showed bilateral infiltration and he was severely hypoxic and hypotensive on admission. He died of multiple organ failure in spite of intensive treatment with mechanical ventilation antibiotics including erythromycin. Legionella longbeachae serotype 1 was isolated from his sputum and was regarded as the etiologic agent. Legionella longbeachae was not isolated from the same type of leaf mold that he used as potting soil. This is the first case of Legionella longbeachae pneumonia from whom the organism was isolated in Japan. PMID- 9847529 TI - [Isolation of methicilline-resistant Staphylococcus aureus (MRSA) from sputum in outpatients]. PMID- 9847530 TI - [Occupational olfactory changes: diagnostic trends]. AB - Olfactory testing has been of minor interest in Occupational Health due to the lack of testing methods able to detect malingering. On the other hand there is evidence that occupational exposure to several, mainly neurotoxic, substances may result in olfactory damage. We have combined three different testing methods in one package in order to assure a forensic-degree level of results. The package consists of: 1. primary neuron functionality testing with a single olfactory stimulant; 2. olfactory-trigeminal discrimination testing with regular sniff test; 3. odor identification score by Doty's UPSIT test. Final judgement of a link between olfactory system impairment and occupational exposure to chemicals requires a good knowledge of the present and past occupational exposure and of the general conditions of the patient. It requires collaboration between the Occupational Health specialist and the expert in Olfactology and may be completed with endoscopy, radiography and other specific controls. We suggest that a more extensive use of appropriate olfactory testing should be established at least for special risk groups of workers. This will not only detect occupational health damage that would otherwise have remained unknown, but can also furnish new information on the neurotoxic effects of many inhalable chemicals. PMID- 9847532 TI - [Review of risks of biological agents and preventive measures to safeguard the health of compost production workers]. AB - A review of studies made in the compost production industry showed the biological agents posing a risk for workers were fungi and thermophile bacteria, gram negative bacteria and endotoxins, with a prevalent inhalation exposure to airborne contaminated dusts. Medical examinations revealed cases of extrinsic allergic alveolitis due to A. fumigatus, and more frequently irritative and infectious disorders occurring especially in conditions of poor environmental hygiene and macroscopic dust pollution. For the evaluation of the air dispersion of microorganisms, which is high in compost transport and turning operations, at present no exposure limit values are available for biological agents; nevertheless, the concentrations measured were often higher than the limit values proposed for other manufacturing sectors by individual authors and by regulatory agencies in Europe, and were comparable to values observed in other industrial settings for which adverse health effects have been shown. Although the number of studies available are few in number, the results suggest that the hazards posed by microorganisms and the poor environmental hygiene conditions often encountered can undoubtedly be a source of risk for workers, which at present is difficult to establish but significant considering the high airborne concentrations of contaminated dust. Besides technical measures to avoid environmental macroscopic dispersion of dusts, measurement of airborne microbiological contaminants is also recommended. Health surveillance needs to be aimed at identifying subjects with hypersusceptibility to the infectious action of the pathogenetic and/or allergenic agents or with hypersensitivity to the same, and also to periodic control of respiratory organs. PMID- 9847531 TI - [First intercalibration test of diffractometric measurements of asbestos in National Italian laboratories]. AB - The Ministry of Health established by law (D.M. 14/5/96) minimum requirements for laboratories intending to perform asbestos analyses, and identified the central facilities that are to coordinate the future quality control programme (based on the kind of techniques used), to which the laboratories must refer in order to continue to operate in the "asbestos" field. Before initiating the quality control programme for the gravimetric technique based on X-ray diffraction measurement, the central facilities decided to carry out a pilot test in a group of sample laboratories in order to assess the current situation in Italy and identify the main problems involved in implementing the programme. In fact, up to the present, no standardized method for the diffractometric measurement of asbestos exists in Italy, so that the various laboratories use different procedures and reference standards. In the pilot test, a group of 15 national laboratories with proven experience performed diffractometric measurements on 3 samples each containing one of the 3 commercial forms of asbestos (chrysotile, crocidolite and amosite). Besides the final result consisting of the percentage by weight compared to the whole sample, information was requested on the measurement procedure and on the reference standards used. The results, although they do not permit definite conclusions to be drawn due to their small number, do provide information that will be of relevance to the organisation of the future quality control programme. Analysis of the data appear to show that neither the measurement procedure used nor the type of asbestos influence the accuracy of the results. The main source of variability and of deviation from the true values seemed instead to depend on the type of standards used and on compatibility, in terms of particle size and comminution technique, between these standards and the samples analyzed. PMID- 9847533 TI - [Considerations on correct diagnosis of atypical electrocardiographic T-wave findings in young subjects]. AB - An analysis is made of the significance of the occasional observation of asymptomatic T-wave changes (inversion). The authors maintain that only an overall evaluation of the anamnestic, clinical and laboratory findings will enable the physician to make a correct diagnosis and exclude from the start, via coordinated protocols, any silent ischaemic disease, and also a series of clinical conditions that the ECG can reveal as this disease. In this connection the authors report a clinical case that is deemed interesting due to the finding of ECG anomalies in an asymptomatic young, athletic subject observed during a periodical medical examination for the purpose of certifying fitness for a specific type of work. PMID- 9847534 TI - Respiratory mucus transportability is impaired in foundry workers: a longitudinal study. AB - Mucus transportability impairment can prolong the permanence of occupational inhalable noxae within ciliated airways. A reliable, non-invasive indicator of mucus transportability is the Normalized Frog Palate Transport Rate (NFPTR). The aim of this 3-year prospective study was to compare NFPTR between a group of 166 foundry workers (E) and a group of 133 power plant workers (NE). In the first and third years of the study, workers underwent: clinical examination, spirometry, NFPTR, chest radiography. In both plants, environmental concentrations of respiratory irritants were well below the limits set by the American Conference of Governmental Industrial Hygienists. Both groups were homogeneous for age and smoking habits. Mean NFPTR was significantly lower in E than in NE in the first and third years of the study, and in smokers in comparison with non-smokers, at the end of the follow-up. NFPTR impairment was significantly associated with occupational exposure in the first and third years of the study. In the third year, a decline in NFPTR was associated with exposure, smoking habits, FVC and FEV1/FVC.100. At the end of the study, the means of FVC, FEV1 and PEF were significantly lower in E. No cases of pneumoconiosis were observed. In this study, low doses of foundry respiratory irritants were associated with impairment of mucus transportability; the consequent slowing of mucociliary clearance increased internal doses of foundry airborne noxae. PMID- 9847535 TI - [Chrono-thermometry in the diagnosis of Raynaud-like vasculopathies caused by prolonged use of vibrating tools]. AB - Raynaud's phenomenon may be secondary to several different pathological conditions. In some cases it is favoured by occupational exposure to vibration of the upper arms. The diagnosis of Raynaud's phenomenon requires clinical information together with specific technical investigations. This report presents the results of 232 chrono-thermometric tests performed on 157 workers exposed to vibrations during the use of different kinds of tools. Seventy-five subjects not exposed to vibrations of the upper arms but with suspected Raynaud's syndrome were also tested. Chrono-thermometry enabled us to distinguish between three different subgroups of subjects exposed to vibrations: 61 workers with "normal" chrono-thermometry, 33 subjects with "pathological" vascular reactivity to cold in some fingers and 63 workers with such reactivity in all fingers. Similarly the 75 patients not exposed to vibration could also be divided into three groups. No statistical difference was found between the chrono-thermometric results obtained from the exposed and non-exposed subjects when divided into the three subgroups. The overall analysis of the combined results of the different kinds of tests (capillaroscopy plus a vascular reactivity test) is discussed. PMID- 9847536 TI - Carbon disulphide as a cause of cerebrovascular disease. PMID- 9847537 TI - [Inter-laboratory comparison of measuring concentration of asbestos fibers less than 5 microM in length using phase-contrast microscopy]. AB - Airborne asbestos fibers at the workplace are usually measured by phase-contrast microscopy (PCM), using the membrane filter method (MFM). Although the MFM has an international standardization, the application of this method can lead to wide differences in results. An inter-laboratory study, involving 19 laboratories, was organized. Two fiber classes were counted: 1) fibers greater than 5 microns in length and less than 3 microns in diameter; 2) fibers less than 5 microns in length and less than 3 microns in diameter. The results of each laboratory were normalised to the mean and their counting performance was compared. The estimated coefficients of variation or relative standard deviation (CV) of the normalised results were fitted assuming a constant component (K) and a component depending on the fiber count (Poisson component: mu-1/2). Fitting showed that the constant K is about 0.36 for counting fibers greater than 5 microns in length and about 0.52 for counting fibers less than 5 microns long. It was shown that counting the latter fiber class is influenced primarily by subjective laboratory-to-laboratory differences. PMID- 9847538 TI - [A punctual errata correction]. PMID- 9847539 TI - Independent reference frames in human spatial memory: body-centered and environment-centered coding in near and far space. AB - Our apparently seamless experience of the spatial environment seems to be derived from information coded across a variety of spatial reference frameworks, each tied to the metric of a different sensory or motor system. A fundamental distinction is that between body-centered and environment-centered reference frameworks. This study reports the first clear evidence of a behavioral dissociation between body-centered and environment-centered coding in human adults. Subjects, seated in a rotating chair with closed eyes, were required to point to remembered, auditorily presented target locations. The subjects were rotated between the presentation and recall of targets. Targets were held stationary with respect to either body-centered or environment-centered spatial coordinates. Prior to recall, subjects were required to point to a series of prelearned distractor locations, which also remained fixed with reference either to the subject's body or to the stationary environment. Memory for the target locations was selectively impaired when distractor locations were specified within the same spatial reference frame as the target, regardless of whether target and distractor locations were near to or distant from the subjects. In contrast, distractor locations specified in a different reference frame from that of the target had either little or no effect on memory for target locations. PMID- 9847540 TI - Verbal and visuospatial short-term memory in children: evidence for common and distinct mechanisms. AB - This study was designed to identify whether verbal and visuospatial short-term memory performance in children is served by common or distinct mechanisms. Five- and 8-year-old children were tested on their verbal recall of spoken letter names and digits, and on their recall of tapped sequences of blocks. The performance of the children on the verbal and visuospatial serial recall tasks was largely unrelated, extending evidence for dissociable memory systems found in adults. Detailed characteristics of recall, such as serial position functions, migration patterns, and distribution of error types, were similar in the tasks requiring recall of letters and of blocks, although order errors predominated in the block but not the letter recall task for the older children. These results appear to reflect the application of common processes specialized for the extraction of serial order information from the phonological and visuospatial components of short-term memory. PMID- 9847541 TI - Name or hum that tune: feeling of knowing for music. AB - This is the first reported research that explores the feeling of knowing (FOK) for musical stimuli. Subjects attempted to recall melodies and titles of musical pieces, made FOK ratings when recall failed, and then had a recognition test. With instrumental music (Experiment 1), more titles were recalled when melodies were given as cues than vice versa. With songs whose lyrics were not presented (Experiment 2), however, more melodies were recalled than were titles. For nonrecalled items, although the overall levels of recognition did not differ, FOK ratings were higher for titles than for melodies in Experiment 1, and the opposite pattern occurred in Experiment 2. In both experiments, the FOK ratings predicted melody recognition more accurately than they did title recognition. PMID- 9847542 TI - Connectedness and part-relation integration in shape category learning. AB - We investigated the role of connectedness in the use of part-relation conjunctions for object category learning. Participants learned categories of two part objects defined by the shape of one part and its location relative to the other (part-relation conjunctions). The topological relationship between the parts (connected, separated, or embedded) varied between participants but was invariant for any given participant. In Experiment 1, category learning was faster and more accurate when an object's parts were connected than when they were either separated or embedded. Subsequent experiments showed that this effect is not due to conscious strategies, differences in the salience of the individual attributes, or differences in the integrality/separability of dimensions across stimuli. The results suggest that connectedness affects the integration of parts with their relations in object category learning. PMID- 9847543 TI - Further evidence for sublexical components in implicit memory for novel words. AB - This study provides further evidence for the notion, proposed by Dorfman (1994), that activation of sublexical components plays an important role in implicit memory for novel words. Priming in an identification task was observed consistently when nonwords were formed out of morphemes; weaker, more variable effects were observed when nonwords were composed of syllables or pseudosyllables. Priming was insensitive to changes in the surface features (type case) of the stimuli and was disrupted by rearrangement of the stimulus components. Results are interpreted in terms of the activation and integration of preexisting sublexical representations and as evidence against the role of newly established episodic or perceptual representations. It is concluded that findings of priming for novel information should not be taken as evidence against an activation view. PMID- 9847544 TI - The decay of short-term implicit memory: unpacking lag. AB - McKone (1995) reported a short-lived repetition priming effect, superimposed on long-lived priming. This short-term implicit memory survived a few intervening items and several seconds for words but decayed precipitously for nonwords, producing a lag x lexicality interaction. Here, mechanisms of decay are studied by disconfounding the time delay and interference components of lag. In Experiment 1, time delay was varied while number of intervening items was held constant, and vice versa. In Experiment 2, priming was plotted as a function of time delay, with and without intervening items. Using a lexical decision task, both experiments found independent contributions of time and interference to the decay of short-term priming. Further, Experiment 2 attributed the lag x lexicality interaction to a particular sensitivity of nonword traces to interference. An illustration of how these effects might arise in the word recognition system is provided. PMID- 9847545 TI - The role of attention and study time in explicit and implicit memory for unfamiliar visual stimuli. AB - The effects of limited attentional resources and study time on explicit and implicit memory were studied using Schacter and Cooper's possible and impossible objects in their recognition and object decision paradigm. In one experiment, when attention at study was limited by a flanking digits procedure, object recognition was diminished but object decision priming for possible objects was unaffected; in another experiment, limiting attention plus reducing stimulus study time impaired object recognition and eliminated object priming. Recognition memory and perceptual priming for previously unfamiliar visual stimuli were both influenced by attention, although to different degrees. The intervening variable of study time determined the degree to which priming was affected by attentional resources. These results support a limited capacity attentional model for both recognition and perceptual priming of unfamiliar visual stimuli, and they highlight the need for assessing the interaction of attentional resources and study time in explicit and implicit memory tasks. PMID- 9847546 TI - A partial matching theory of the mirror effect in immediate probed recognition. AB - A set of experiments on immediate probed recognition of digit triples is reported in which the variables were list length (five, six, seven, or eight triples), the probability that a probe was old (.33, .5, or .67), and whether the digit triples were presented with an auditory component or articulatory suppression. Previous work had suggested that the false alarm (FA) rate in this paradigm was lower when auditory information was available than when it was not; this observation had led to the development of the partial matching theory of immediate probed recognition, according to which FAs could arise not only as a result of unlucky guesses but also when new probes shared a first digit in common with a partially retained target triple. It was argued that partial memory representations were less likely following auditory presentation than following articulatory suppression. Partial matching theory is contrasted with the rational response theory, according to which all FAs are unlucky guesses; partial matching theory gave a better account of the present experimental data than did rational response theory. However, a logical relationship between the two theories was suggested, a consequence of which was that rational response theory could be modified to include partial matching in such a way as to account for mirror effects, not only in unusually difficult immediate probed recognition tasks, but also in the more commonly studied mixed test list paradigm involving words of high or low frequency. PMID- 9847547 TI - An examination of the distinction between nouns and verbs: associations with two different kinds of motion. AB - Four experiments provide evidence that people are biased to associate particular types of motion with nouns and different types of motion with verbs. Novel nouns and verbs were related to two types of motion: (1) path, or the direction of motion of one character with respect to the other character, and (2) movement orientation, or the direction a character was facing as it moved. Subjects associated verbs more strongly with path than with movement orientation. In contrast, they associated nouns more strongly with movement orientation than with path. Movement orientation was associated with both object categories and verbs, inconsistent with a complete division of labor between these two types of categories. These results are consistent, however, with the notion that people are biased to associate verbs with relations between objects, whereas they are biased to associate object categories with motions defined with respect to the object carrying out those motions. PMID- 9847548 TI - Assessing the accuracy of conflicting autobiographical memories. AB - We examined criteria that rememberers and neutral audiences use to validate conflicting memories. In Experiment 1, rememberers described an incident that they recalled differently from someone else from their own and the other person's perspective. Rememberers and audiences then evaluated the accuracy of statements in both accounts, explained their accuracy ratings, and appraised the qualities of the memories. In Experiment 2, dyads who possessed conflicting memories of a shared past episode evaluated each other's recollections. In both studies, rememberers rated their own recollections as more accurate than other people's and based their judgments primarily on the internal coherence of the memories. Rememberers used consensus more frequently and sensory detail less often to evaluate other people's memories than their own. Audiences (Experiment 1) reported examining the consistency between the accounts and their own experiences; they also used consensus and normative behavior to a greater extent than rememberers. In both studies, rememberers judged their own accounts to be more detailed and coherent. Their ratings of detail and coherence predicted their ratings of accuracy. PMID- 9847549 TI - More than meets the eye: context effects in word identification. AB - The influence of semantic context on word identification was examined using masked target displays. Related prime words enhanced a signal detection measure of sensitivity in making lexical decisions and in determining whether a probe word matched the target word. When line drawings were used as primes, a similar benefit was obtained with the probe task. Although these results suggest that contextual information affects perceptual encoding, this conclusion is questioned on the grounds that sensitivity in these tasks may be determined by independent contributions of perceptual and contextual information. The plausibility of this view is supported by a simulation of the experiments using a connectionist model in which perceptual and semantic information make independent contributions to word identification. The model also predicts results with two other analytic methods that have been used to argue for priming effects on perceptual encoding. PMID- 9847550 TI - Comparing naming, lexical decision, and eye fixation times: word frequency effects and individual differences. AB - Performance on three different tasks was compared: naming, lexical decision, and reading (with eye fixation times on a target word measured). We examined the word frequency effect for a common set of words for each task and each subject. Naming and reading (particularly gaze duration) yielded similar frequency effects for the target words. The frequency effect found in lexical decision was greater than that found in naming and in eye fixation times. In all tasks, there was a correlation between the frequency effect and average response time. In general, the results suggest that both the naming and the lexical decision tasks yield data about word recognition processes that are consistent with effects found in eye fixations during silent reading. PMID- 9847551 TI - Contrasting effects of age of acquisition and word frequency on auditory and visual lexical decision. AB - In four experiments, we examined the effects of frequency and age of acquisition on auditory and visual lexical decision. Word frequency affected visual, but not auditory, lexical decision speed (Experiments 1 and 3). Age of acquisition affected lexical decision speed in both modalities (Experiments 2 and 4). We suggest that previous reports of effects of frequency on auditory lexical decision may be due to a confounding of frequency with age of acquisition, and we discuss the implications of these findings for theories of auditory and visual word recognition. PMID- 9847552 TI - Context effects in visual word recognition: lexical relatedness and syntactic context. AB - In three experiments, we investigated how associative word-word priming effects in German depend on different types of syntactic context in which the related words are embedded. The associative relation always concerned a verb as prime and a noun as target. Prime word and target word were embedded in visually presented strings of words that formed either a correct sentence, a scrambled list of words, or a sentence in which the target noun and the preceding definite article disagreed in syntactic gender. In contrast to previous studies (O'Seaghdha, 1989; Simpson, Peterson, Casteel, & Burgess, 1989), associative priming effects were not only obtained in correct sentences but also in scrambled word lists. Associative priming, however, was not obtained when the definite article and the target noun disagreed in syntactic gender. The latter finding suggests that a rather local violation of syntactic coherence reduces or eliminates word-word priming effects. The results are discussed in the context of related work on the effect of gender dis-/agreement between a syntactic context and a target noun. PMID- 9847553 TI - The influence of semantic fit on on-line sentence processing. AB - The interactive influence of verb complement preferences and noun phrase semantic fit on resolution of temporary syntactic ambiguity was investigated in an eye movement experiment. The present semantic fit manipulation included noun phrases that fit well as direct objects of the verbs that they followed and noun phrases that were possible but less likely direct objects of the verbs in question. This contrasted with existing research on the use of verb complement preferences and semantic fit during sentence processing, in which processing of noun phrases that are possible direct objects has been compared with processing of noun phrases that are not possible direct objects of the verbs that they follow. Verb complement preference information and noun phrase semantic fit interacted at early stages of on-line sentence processing. Implications of these results for interactive and structural models of sentence processing are discussed. PMID- 9847554 TI - Focus as a contextual priming mechanism in reading. AB - In two experiments, we found that readers are sensitive to manipulations of syntactically marked focus and that focus is an effective message level contextual priming mechanism. Changes in focus resulted in changes in sentence context effects on subsequent target word processing. This was demonstrated in latency to name the target word (Experiment 1) and in initial looking time on the target in silent reading (Experiment 2). Experiment 2 also revealed direct effects on the focused items, as readers made fewer regressions and spent less total time on a word that was focused than when it was not focused. However, no initial processing time effects were found on the focused word. PMID- 9847555 TI - The influence of types of character on processing background information in narrative discourse. AB - The background sentence The air was cold and clammy depicts setting information which may or may not be of significance to a given character in a narrative. We tested the hypothesis that such information is processed with respect to the main character in a narrative rather than with respect to a secondary character. In Experiment 1, subjects making attributions of awareness of such background states were more likely to attribute such awareness to main than to secondary characters. In Experiment 2, using self-paced reading, we showed that such information is more important for main than for secondary characters during reading. Thus, unattached background information is processed with respect to main characters. This bias provides a source of control over the processing necessary to establish a coherent representation during the reading of narratives. Experiment 3 eliminated the possibility that the effects were due to generally shallower processing of sentences centered on secondary characters. PMID- 9847556 TI - Property instantiation in conceptual combination. AB - In four experiments, I examined how a property in one concept is transferred to a second concept during conceptual combination. The results suggest that people instantiate properties: that is, they use a specific representation of a property in the modifier concept to construct a new version of that property that is specific to the combination. If people are instantiating properties, then the modifier property should match its counterpart in the combination to the extent that the modifier and head noun are similar. This observation leads to a variety of predictions (supported by the experiments) about interpretations of similar and dissimilar combinations and about plausibility, preference, and similarity judgments associated with such interpretations. The results argue against an alternative view of transfer that posits that, in general, abstract representations of properties are copied from one concept to another. In this paper, I describe various processing accounts of instantiation and discuss the implications of the instantiation view for theories of metaphor, conceptual combination, and induction. PMID- 9847557 TI - [Source of information on salt intake in Japan and its influence on the understanding of nutrition guides on salt intake]. PMID- 9847558 TI - [Outcomes of stroke survivors in Yamagata Prefecture]. AB - PURPOSE: To clarify the long-term outcomes of Stroke survivors registered for the first onset of Stroke in Yamagata Prefecture and to find out problems in community-based-rehabilitation (CBR). SUBJECTS AND METHODS: The present study was performed using stroke survivors as of September 1, 1991 as subjects from residents registered for the onset of Stroke in 1985 and 1989. The subjects were composed of 1,013 residents registered in 1989 (2 years after onset) and 626 registered in 1985 (6 years after onset). RESULTS: The percent of functionally independent Stroke survivors at 2 and 6 years (indicated in parentheses) after onset is shown by ADL items as follows: 82% (81%) for urination, 78% (78%) for eating, 78% (78%) for walking, 76% (78%) for dressing, and 66% (64%) for bathing. The lowest percent was seen in Bathing. Percentage of cases maintaining the ability to have functionally-independent ADL for all items examined (expressed as persons independent for personal care) were 62% (60%). Of the the cases 91% (91%) resided at home (their own houses or relatives' houses). With regard to overall locomotion, an item used to evaluation the range of going out doors, 45% (44%) could go out alone to visit neighbors or use public transportation. Among persons independent for personal care, 99% (98%) were living at homo, 70% (69%) used public transportation, 23% (23%) went out alone to visit neighbors and 7% (8%) did not go out. CONCLUSION: The results of the present research indicate a great in CBR for that bothpsychological and social health approaches in cooperation with public health centers, medical centers, and welfare agencies not only for persons with decreased ADL, but also for those maintaining high activity potential. PMID- 9847559 TI - [Tuberculin sensitivity among school children in Niigata Prefecture, Japan]. AB - Tuberculin sensitivity was studied in 668 apparently healthy schoolchildren six to seven years of age vaccinated B.C.G. before the age of one. Sensitivity was assessed using the standard dose of PPD and reactions with an area of erythema of 10 mm diameter or more were considered positive. The results were as follows: 1) The frequencies of tuberculin-positive children were 97 (31.3%) of 310 subjects in the fishing area, 21 (9.9%) of 213 in the farming area, and 35 (24.1%) of 145 in the intermediate zone. 2) Prevalences of the positive children by obesity index were 28 (36.8%) of 76 in the large, 72 (24.5%) of 294 in medium, and 53 (17.8%) of 298 in the small, and the decreasing tend in frequencies among these three classes by obesity index were statistically significant (p < 0.01). 3) Gender differences in frequency of tubereulin-positive children were not clearly detected when a comparative study grouped by obesity index and area was carried out. These observations suggest that tuberculin sensitivity in children may be influenced by the size of body mass and also characteristic factors, including diet, in their residential districts. PMID- 9847560 TI - [Prevalence and a drug use development model for the study of adolescent drug use in Japan]. AB - The purposes of this study were to estimate the prevalence of drug use, to examine the correlation between drug use and drinking/smoking in Japan, and to test a developmental model of adolescents' drug use based on three basic theories of deviance: Strain, Social control, and Differential association. The survey was conducted from April to May, 1996, in 14 public high schools in Tokyo, using anonymous self-reporting. A total of 4,171 (99.45%) students responded to our questionnaire. It included central concept items in the three deviant theories, beliefs about the ill effects of drugs, sex, grades, etc. The following findings were obtained: 1) Among the subjects, 255 students (6.1%) reported drug use within the past year. Of these drug users, 73.3% used drugs soon after being "tempted" by friends. Responding to "When was it.", as a first year junior high school was the most common first experience, and the next most common was during the sixth year of elementary school. 2) Drinking and smoking appeared to be gateway drugs for adolescents in Japan as has been shown in the United States. 3) Multiple regression and logistic multiple regression analyses suggested that differential association variables were far more powerful predictors of adolescent drinking, smoking and drug use than either the control or strain variables. Results provided modest support for the Differential Association Theory as an explanation of drug use. 4) LISREL's goodness-of-fit statistic indicated a much better fit between the model and the data. (CN: 282, GFI: 0.967, AGFI: 0.941). These findings show two processes by which adolescents become involved in drug use. Strain and Social Control do not directly affect drug use. However, Social Control is important because it works indirectly, through Differential Association resulting in drug use. Second, though weaker, Differential Association appears to lead to dangerous beliefs in drug use followed by actual drug use. PMID- 9847561 TI - [Homebound elderly in a Japanese community: related factors and change of mobility]. AB - The aim of this study was to identify the prevalence of homebound elderly (defined as people whose daily activities were limited to their home) and factors related to it among 300 community elderly residents aged 60 and over in Yamagata city. In 1995, the baseline survey was performed and the follow-up survey was conducted one year later. Subjects were divided into two groups according to the extent of their daily activities: the non-homebound group (defined as people whose daily activities extended into their community) and the homebound group. The main results were as follows; 1. The prevalence of homebound elderly was 7.7% in 1995. 2. Chi-square test or t test was performed to examine the relationship between homebound and various factors. Significant factors were age, history of hypertension, history of mental disease, incompetence of ADLs (walking, eating, toileting, bathing, dressing), interpersonal dependency, subjective health, 'ikigai' (meaningfulness of life), life style (cooking, cleaning, reading newspaper or magazine, watching TV, exercise, associate with friend) and TMIG (Tokyo Metropolitan Institute of Gerontology) index of competence. The present study reveals that daily activities in community elderly residents is related to not only physical factors but also psychosocial factors. Using the significant variables in univariate analysis, multiple logistic regression analysis controlling for age was performed. Significant factors for homebound were incompetence of ADLs (walking, toileting), subjective health and TMIG index of competence. 3. Three out of 214 non-homebound elderly persons in 1995 changed to homebound in 1996. PMID- 9847562 TI - [Assessment method for supporting programs for social activities of the elderly by city, town and village governments in Japan]. AB - OBJECTIVES: The authors developed a questionnaire to assess the activities of supporting programs for social activities for the elderly conducted by city, town and village governments, based on the number of such programs. This study aims to examine the validity of the questionnaire. METHODS: We conducted a study of 145 cities, towns and villages from two prefectures in Japan using the questionnaire. The questionnaire asks government officers whether they had each of 32 programs, and the number of programs in total, as well as each of four domains of social activities (i.e., employment, social participation/volunteering, education/training, individual activities) were calculated. They were also asked to classify each of the programs into one of four domains of social activity to which the programs were most related; they were asked to rate how active the programs were for each domain in their community. To examine the validity of the questionnaire, the following indicators were assessed: (1) proportion of reported programs which were not covered by the questionnaire; (2) concordance of classification of programs into four domains of social activity with those judged by government officers; (3) correlations between number of programs and self rated activity levels by government officers; and (4) correlations of number programs with characteristics of cities, towns or villages. RESULTS: The results indicated that the list of programs covered most of the programs. Proposed classification of programs into four domains of social activities which they are most related to agreed with judgments by officials. The number of programs positively correlated with self-rated activity levels by officers for employment and education/training domains in both prefectures; and for social participation/volunteering and individual activities in one of the prefecture. The numbers of programs for employment of the elderly positively correlated with the total population of the community, number of staffs, proportion of people employed in the tertiary industries and total budget for social welfare of the elderly; it negatively correlated with proportion of people employed in the primary industries. CONCLUSION: It is suggested that the list of programs in the questionnaire is applicable and assessment based on the number of programs by four social activity domains is valid to some extent in either prefecture. The questionnaire can be improved further, e.g., by adding other programs to the list. PMID- 9847563 TI - [Relationship of utilization rate of health examination under the Elderly Act to medical expenditure and days of medical care for the elderly]. AB - The aim of this study was to investigate the relationship of utilization rate of health examination to medical expenditure and days of medical care per insured person under the Elderly Act. The factors related with the utilization rate of health examination were also analyzed by multiple regression analysis. The material used in this analysis was the 1993 national health insurance data for all of the 3,252 municipalities of Japan. The main results were as follows: 1. The average medical expenditure and days of medical care among the municipalities declined in the increasing order of the three categories of utilization rate of health examination; municipalities with utilization rate of less than 30% being the highest, and those with 50% or more being the lowest. 2. The average medical expenditure and days of medical care among the municipalities with community health centers were smaller than those without. 3. Multiple regression analysis showed that utilization rate had a positive relationship to percentage of elderly, presence of community health center, public financial power index, and number of public health nurses per resident. It had a negative relationship to the population and the number of clinical beds per resident. The number of public health nurses per resident was the most significant factor related with the utilization rate (standardized partial regression coefficient = 0.32). These results suggest that increasing the utilization rate of health examination and the number of community health centers could serve to lower medical expenditure and days of medical care through the promotion of the health services provided by municipalities. PMID- 9847564 TI - [Effect of home visit dental health services]. PMID- 9847565 TI - [Types and dimensions of mineral fibers in the atmosphere and from lungs of residents in the urban area]. AB - OBJECTIVE: The objective of this study was to estimate the types and origin of the mineral fibers in the lungs of residents in the urban area. METHODS: Mineral fibers in the atmosphere from 21 urban areas were analyzed by direct transfer analytical transmission electron microscopy (ATEM). Lung tissues of 36 urban residents without occupational asbestos exposure were also analyzed by ATEM after low temperature ashing. Measurement of dimensions of all types of mineral fibers, identification of asbestos, and classification of nonasbestos fibers according to the combination of main elements were carried out. RESULTS: 1. Chrysotile, amosite, actinolite, tremolite and 11 types of nonasbestos fibers were found in both the lung tissues and the atmospheric samples. The fibers, mainly composed of Al and Si, contributed 16.3% and 62.3% of the atmospheric samples and lung tissues respectively. On the other hand, the proportion of the fibers mainly composed of S and Ca were 43.7% and 0.0% in the atmosphere and lung tissues respectively. Those fibers mainly composed of P and Fe were 0.4% and 18.4% in the atmosphere and lung tissues respectively. 2. Fifteen types of nonasbestos fibers were found exclusively in the atmosphere. The average proportions for ten of these were less than 0.5% each. 3. Crocidolite was found exclusively in the lung tissues. 4. No significant difference in the geometric mean of length and diameter of asbestos was found between the atmospheric samples and lung tissues. Some types of nonasbestos fibers had significantly larger length and diameter in the atmospheres than in the lung tissues (p < 0.01). CONCLUSION: The results suggest that mineral fibers found in lung tissues could be classified into three groups on the basis of their behavior. 1) Some fibers, such as amphibole asbestos and fibers composed of Al and Si, are inhaled from the atmosphere and remain in the lung tissues for a long time. 2) Some fibers, such as those composed of S and Ca, are inhaled from the atmosphere and eliminated from the lung tissues within a relatively short time. 3) Some fibers found more in the lung tissues than in the atmospheres, such as those composed of P and Fe, are suspected to be generated in the lungs. PMID- 9847566 TI - Nedaplatin and 5-FU combined with radiation in the treatment for esophageal cancer. AB - A series of patients with esophageal cancer was treated with chemotherapeutic regimens of the new antitumor platinum preparation nedaplatin plus 5-FU in combination with radiation therapy, and the therapeutic responses, side effects, and complications were clinically assessed. There were 2 patients with a complete response and 11 patients with a partial response, hence, a response rate of 76.5%. Major adverse reactions were those of hematological toxicity and included leukopenia (13 patients, 76.5%), thrombocytopenia (8 patients, 47.1%), and lowered serum hemoglobin concentration (9 patients, 52.9%). The leukopenia and thrombocytopenia, though of a grade 3 severity in 3 and 2 patients, respectively, subsided spontaneously in all affected cases. Gastrointestinal adverse reactions were mild and included appetite loss in 7 patients (41.2%) and nausea in 2 patients (11.8%). The only abnormality in renal function observed was a slight elevation of serum creatinine in one patient. The combined therapy of chemotherapy with nedaplatin and 5-FU plus radiation produced a high response rate in the treatment of carcinoma of the esophagus and was associated with reduced gastrointestinal and renal toxicity. The results indicate the combined therapy with nedaplatin to be clinically useful. PMID- 9847567 TI - A surgical case of acute three-channeled aortic dissection in Marfan syndrome. AB - We surgically treated a 35-year-old male with acute 3-channeled aortic dissection in Marfan syndrome. He had acute type A aortic dissection, and underwent Bentall's type operation, simultaneous graft replacement of the ascending aorta and total aortic arch. Pain recurred 5 years and 9 months after the first operation. CT scan showed two adjacent false lumens in the descending aorta. The morphology of the first and second dissections was Stanford type A + B. The second dissection was acute. In the second false lumen, a re-entry formation was observed in the abdominal aorta. Because severe pain was persistent, we immediately replaced the descending aorta using a femoro-femoral partial cardiopulmonary bypass. The patient was doing well and was discharged. When pain recurs in a Marfan patient with an aortic dissection, a 3-channeled aortic dissection should be suspected, and we recommend emergency surgery. PMID- 9847568 TI - Successful surgical management of penetrating cardiac injury. AB - A 60-year-old Asian man who suffered penetrating thoracoabdominal injuries was admitted to our hospital in a state of shock. Because the results of a chest computed tomogram (CT) strongly suggested a left ventricular injury, an emergency thoracotomy was performed. A laceration of the left ventricle (3 cm) was sutured and was closed without cardiopulmonary bypass, and coexisting lacerations of the superior mesenteric artery (SMA) and small bowel were surgically repaired. During the procedure, cardiac arrest occurred, but the patient recovered without any apparent neurologic deficit. Postoperative examinations using echocardiogram, CT, and cardiac catheterization revealed a delayed rupture of the left ventricle. On the 28th day after surgery, he was transferred to another hospital for elective cardiac surgery. Primary management of penetrating cardiac injury is discussed in this report. PMID- 9847569 TI - Experience with thoracoscopic surgery for primary bronchial stump fistula after pneumonectomy. AB - After a left pneumonectomy, thoracoscopic closure with fibrin glue was performed for a fistula on the bronchial stump and the postoperative state progressed favorably thereafter. In this paper, we report on this successful case. CASE: A 61 year-old male, who underwent a left pneumonectomy on January 17, 1996 for pulmonary carcinoma (T 3 N 1M 0 stage III A). The bronchial stump was covered with anterior serratus muscle flap. On April 1 (the 76th postoperative day), after two courses of Carboplatin and Vindesine treatment, the patient suddenly developed a fistula on the bronchial stump. Bronchofiberscopic closure with fibrin glue was attempted, but failed to close the fistula. Thoracoscopic surgery was then performed on May 15 (the 45th day after the onset of the fistula). After the intrathoracic opening of the fistula was found with a contrast medium, fibrin glue was injected to fill up to the bronchial stump, and communication with the thoracic cavity was blocked. Owing to coverage with a myocutaneous flap, the patient's general postoperative state remained relatively stable. Thoracoscopic surgery is useful as a treatment for some cases of bronchial stump fistula after pneumonectomy. PMID- 9847570 TI - [Blood conservation effect and safety of shed mediastinal blood autotransfusion after cardiac surgery]. AB - Autotransfusion of shed mediastinal blood after cardiac surgery has been used to reduce risks related to homologous blood transfusions. To document the efficacy and safety of autotransfusion, we compared clinical findings of 80 patients receiving shed mediastinal blood (autotransfusion group) with those of the control group of 52 patients. The amount of the autotransfusion was limited to 800 ml, given the potentially harmful effects of shed blood transfusion. The mean transfused shed volume was 314 +/- 236 ml (S.D.). The serum levels of FDP-E, D dimer and TAT after autotransfusion were higher in the autotransfusion group than in the control group (p = 0.01, p = 0.0004, p = 0.001, respectively). However, postoperative blood loss and the rate of reexploration for bleeding were similar in the two groups. The patients receiving blood products were fewer in the autotransfusion group than those in the control group (21% vs 44%; p = 0.005). Autotransfusion did not increase postoperative complications, including infection. Thus, although autotransfusion of mediastinal shed blood has the potential to affect hemostasis, unless the amount of autotransfusion exceeds 800 ml, it appears that this method is clinically safe and effective as a mean of blood conservation. PMID- 9847571 TI - [Coronary artery bypass grafting for cases which were not amenable to preoperative left ventriculogram]. AB - We evaluated the coronary artery bypass grafting (CABG) for the cases which were not amenable to preoperative left ventriculograms (LVG). Between June 1970 and March 1996, we had 2045 cases of isolated CABG, and 52 cases of which were not amenable to preoperative LVG. The reasons of no preoperative LVG were during cardiopulmonary resuscitation (CPR) in 4 cases, shock state in 14, NYHA class IV or CCS class IV in 26, angina attack after coronary angiography in 3 and so on. Preoperative diagnoses were acute myocardial infarction in 28, angina after infarction in 13, unstable angina in 9 and effort angina in 2. Preoperative conditions were reoperation in 6, insertion of intra-aortic balloon pumping in 46, chronic renal failure in 15 (hemodialysis in 4) and arrhythmia (VT or Vf) in 12. Concerning the coronary lesions, left main trunk and triple vessel diseases occupied 75% of all. Surgical deaths were 5 cases (9.6%) and hospital deaths were 4 cases (7.7%). The all cases which needed preoperative CPR were died with brain injury. The result of the cases which were not amenable to preoperative LVG were worse than the cases with ischemic cardiomyopathies. In conclusion, no preoperative LVG is one of the highest risk factors of all. PMID- 9847572 TI - [Clinical results of endovascular stent graft repair for fifty cases of thoracic aortic aneurysms]. AB - Between February 1995 and December 1997, 50 cases (55 lesions) of thoracic aortic aneurysms including 20 cases of aortic dissections were treated with an endovascular technique using the stent grafts. All patients were treated in the operating room under general anesthesia and the stent grafts were implanted through 18 Fr. or 20 Fr. sheaths via femoral arteries under fluoroscopic guidance. The stent graft was composed of several units of self-expanding stainless-steel Z stents covered with an ultra-thin polyester fabric. Stent graft deployment was technically successful in 53 of 55 lesions (delivery success rate: 96.4%). Exclusion of the aneurysms and entry closing without endoleak were achieved within two weeks after the operation in 43 of 53 lesions (initial success rate: 81.1%). Endoleak was found in 10 lesions (minor endoleak: 8 and major endoleak: 2 lesions). Two patients died in the periopertive period of delivery failures as injury to external iliac artery and damage to the delivery sheath caused by tortuous and narrow access routes. Endovascular stent graft repair of thoracic aortic aneurysms is minimally invasive operation in comparison with conventional surgical graft replacement with extracorporeal circulation. These early results suggest that the stent graft repair is possibly safe and useful treatment for the patients of thoracic aortic aneurysms especially in high risk patients. However, careful long-term follow-up is necessary to prove the value and the effects of this endovascular treatment and improvement of the stent graft system and technical training of endovascular surgery for operators are required to reduce the delivery failure and to determine the stent graft repair is reliable treatment. PMID- 9847573 TI - [An experimental study of local chemotherapy for metastatic lung tumor--isolated lung perfusion and pulmonary artery infusion]. AB - PURPOSE: We evaluated the efficacy of local chemotherapy using isolated lung perfusion (ILP) and pulmonary artery infusion (PAI) in vivo to improve prognosis for patients with metastatic lung diseases. MATERIAL AND METHODS: Male F344 rats were used. The left lung was isolated and chemotherapeutic agents were administrated to the isolated lung by the ILP or PAI technique. Agents used were cisplatin (CDDP) and fluorouracil (5-FU), and their levels in the perfused lung and serum were measured (renal levels were also measured in CDDP groups). The results were compared with those obtained following intravenous administration (i.v.). In the ILP group, the left pulmonary artery and vein were clamped during perfusion. The agents were infused from the pulmonary artery, and the perfusate was collected from the pulmonary venotomy site following flushing with saline before declamping. In the PAI group, the left pulmonary artery was clamped and perfused with 0.2 ml of the agents, pulmonary arterial flow was occluded for 20 minutes, and perfusate was not collected. Rats were sacrificed 5 minutes after declamping in both groups, and the perfused lung and blood (and also kidneys in the CDDP group) were collected for pharmacokinetic examination. Histological examination of the perfused lung was also performed. RESULTS: In the ILP group treated with 1 mg of CDDP and PAI group treated with 0.1 mg of CDDP administration, the lung CDDP levels were significantly higher and the serum and renal CDDP levels were significantly lower than those in the IV group. In the ILP group treated with 150 mg of 5-FU and PAI group treated with 10 mg of 5-FU, the lung 5-FU levels were significantly higher than those in all IV groups and the serum 5-FU levels were lower than those in the IV group treated with 10 mg and 15 mg of 5-FU. The bronchopulmonary construction was histologically maintained in the perfused lung in each group. CONCLUSION: We concluded that local chemotherapy using ILP or PAI could be a safe and effective choice of therapy, and this method may be clinically applicable. PMID- 9847574 TI - [Coronary artery bypass graft surgery in dialysis patient]. AB - To determine the operative outcome of coronary artery bypass graft surgery (CABG) for severe coronary artery disease in long-term hemodialysis patients, we analyzed a group of 16 patients who underwent CABG over a ten-year period in our institution. Hospital mortality was 12.5% (2 of 16 patients). These two patients died of ischemic colitis and perioperative myocardial infarction, respectively. There were five late deaths: one patient died from myocardial infarction, one from uremia, one from gastro-intestinal bleeding, one from gastric cancer and one from unknown cause. There were four significant postoperative complications (morbidity 25%), consisted of one pulmonary tuberculosis, one sternal dehiscence secondary to mediastinitis, one mediastinal hematoma secondary to late bleeding from the LITA dissection area and one A-V shunt trouble. Graft patency rate within the first two months was 93% (30 to 42 in 13 patients). Hospital survivors experienced complete relief from angina. Actuarial survival was 68.8% at 3 years, 57.3% at 5 years and 28.6% at 7 years. This rate is not significantly different from the survival of all dialysis patients, but seems to be better than that of dialysis patients with not operated coronary artery disease. We concluded that CABG in dialysis patients can be accomplished with acceptable morbidity and mortality and effective relief of symptoms. PMID- 9847575 TI - [Video-assisted thoracoscopic surgery in spontaneous hemopneumothorax]. AB - We retrospectively studied the safety and utility of video-assisted thoracoscopic surgery (VATS) in the treatment of spontaneous hemopneumothorax. Of 128 cases of spontaneous pneumothorax operated on our hospital from April 1988 to October 1997, hemopneumothorax developed in 8 cases (2 cases treated by thoracotomy and 6 by VATS). In all 8 cases, bleeding points and pulmonary bullae were easily found and hemostasis and resection of pulmonary bullae conducted quickly and safely. Two cases of VATS involved elective surgery. Of surgical emergent cases, the duration from visit our hospital to operation and surgical duration in VATS were almost as long as those in thoracotomy. The mean duration of postoperative chest drainage and postoperative hospital stay in VAST were less than in thoracotomy except for a VAST case with persistent air leakage. Blood loss from onset to operation and blood transfusion for VATS were almost equal to thoracotomy. Postoperative duration of analgesic use for VATS were shorter than that for thoracotomy. The VATS case with persistent air leakage should be necessary to reinforce the pulmonary stapled line or to convert to thoracotomy. In all cases, residual hematoma was found in the thoracic cavity. We conclude that early surgical repair should be performed once spontaneous hemopneumothorax is diagnosed and confirmed, and that VATS may be the first choice of surgery because it provides a better view and more facilitated manipulation during surgery than thoracotomy, and is a safe, nonaggressive therapeutic option. PMID- 9847576 TI - [An experimental study on the occurrence of brain edema after retrograde cerebral perfusion]. AB - To assess the safety of retrograde cerebral perfusion, the occurrence of brain edema after this procedure was investigated. Twenty-eight adult mongrel dogs were divided into three groups that underwent the following treatments: antegrade perfusion (group 1, n = 9); retrograde perfusion alone (group 2, n = 11); or tetrograde perfusion with drugs (manuitol, thiopental sodium, and methylprednisolone; group 3, n = 8). After 90 minutes of cerebral perfusion at 20 degrees C of the pharyngeal temperature, evans blue (EB) was administered to check for disruptions of the blood-brain-barrier (BBB) and brain tissue water content was measured. Intracranial pressure after cerebral perfusion was markedly higher in group 2 than in group 1 (26.4 +/- 9.4 vs. 11.2 +/- 3.6 mmHg), and brain tissue water content was also significantly higher in group 2 than in group 1 (80.7 +/- 2.0 vs. 77.8 +/- 0.9%). These data suggested that brain edema was more prominent after retrograde perfusion than after antegrade perfusion. The extent of EB to brain tissue was greater in group 2 than in group 1 (169.8 +/- 97.7 vs. 54.7 +/- 31.5 micrograms/dl). The BBB was highly disrupted in group 2 and vasogenic edema appeared after retrograde cerebral perfusion. Maximum intracranial pressure, brain tissue water content and EB concentration were significantly lower in group 3 than in group 2, and did not differ significantly between group 3 and 1. Administration of pharmacologic agents suppressed edema formation and extravasation of EB. We conclude that 90 minutes of retrograde cerebral perfusion at 20 degrees C of the pharyngeal temperature causes brain edema and disrupts the BBB in a manner different from that associated with antegrade perfusion. Mannitol, thiopental sodium, and methylprednisolone prevent these phenomena, indicating that pharmacologic intervention may improve the safety of retrograde cerebral perfusion. PMID- 9847577 TI - [A case of complete circumferential aortic rupture due to blunt chest trauma]. AB - A case of complete circumferential rupture of the thoracic aorta due to blunt chest trauma is presented. A 30-year-old man was admitted after a traffic accident. The admission chest X-ray film demonstrated mediastinal widening. About 6 hours after the traffic accident, chest CT scanning demonstrated mediastinal hematoma, left pleural effusion and partial aortic dissection. Diagnoses of aortic rupture and liver injury were made, and surgery for the thoracic aorta was performed immediately. The aorta was found to be completely disrupted for the length of 2 cm, and a vascular prosthesis was interposed between the two ends of the aorta under partial cardiopulmonary bypass. We conclude that the enhanced chest CT scanning is helpful for diagnosis, and that if other organ injuries are not severe, the emergency operation should be performed. PMID- 9847578 TI - [Acute type A aortic dissection with leg ischemia]. AB - The incidence of lower extremity ischemia secondary to acute aortic dissection is relatively low, however, the presenting symptoms are variable in term of severity. We report here in two cases of such circumstances who were successively differently treated. Case one was a 60 years old male presented with severe left leg pain. Even after the initiation of cardiopulmonary bypass, the leg ischemia did not improve, therefore selective leg perfusion was additionally performed through direct left femoral artery cannulation. The surgery toward dissection was completed by mean of simultaneous graft replacement of ascending aorta and aortic arch. The leg ischemia after the aortic procedure however had persisted, femorofemoral bypass was created to relieve the mal-perfusion. Case two was a 37 years old male admitted with severe left leg pain associated with sensory-motor nerve dysfunction with muscle rigidity. In this particular patient, femoro femoral bypass was firstly reconstructed as the mean of leg salvage procedure. After we learned there was no serious reperfusion symptom manifested, we performed radical surgery toward the aorta. We believe that the decision making of surgical treatment for acute type A dissection complicated with the presence of lower extremity ischemia is based on the severeness of mal-perfusion. PMID- 9847579 TI - [Aneurysm of the membranous ventricular septum with ventricular septal defect, mitral and tricuspid insufficiency]. AB - A seventy-year-old man was admitted at our hospital because of dyspnea. Echocardiogram and left ventriculogram showed an aneurysm formation of the membranous ventricular septum and small left-to-right shunt through ventricular septum defect and also severe mitral and tricuspid insufficiency. Operation was performed after medical therapy for congestive heart failure. During operation, mitral leaflets showed no organic lesions nor prolapse, but the annulus was dilated. The cause of mitral insufficiency, we thought, might be congenital, and the annulus dilatation was caused of mitral insufficiency, we thought, might be congenital, and the annulus dilatation was caused to produce tricuspid insufficiency secondary. The ventricular septal communication became small (diameter; 5 mm) and was associated with aneurysm formation of the remaining portion of the membranous septum. And the aneurysm, protruding to the septal leaflet of tricuspid valves, enhanced tricuspid insufficiency. It was reported by many authors that the aneurysm formation was related to spontaneous closure of ventricular septal defect. Patients with small ventricular septal defect, without any symptoms, must be followed intensively, or they might get cardiac complications, such as arrhythmia, right ventricular outflow obstruction, tricuspid insufficiency, and so on. PMID- 9847580 TI - [A case of emergency surgery for acute mitral regurgitation due to complete papillary muscle rupture as complication of acute inferior myocardial infarction]. AB - We experienced a case with acute mitral regurgitation caused by complete posterior papillary muscle rupture as complication of acute inferior myocardial infarction, who underwent successfully emergency operation of mital valve replacement and coronary revascularization in acute stage. A 64-year-old woman developed sudden cardiogenic shock shortly after the onset of acute inferior myocardial infarction. The diagnosis of acute inferior myocardial infarction was based on the electrocardiographic findings. Under IABP support, preoperative coronary angiography visualized total occlusion of segment 3 of the right coronary artery, and preoperative left ventriculography showed akinesis of inferior wall and severe mitral regurgitation. At 6 hours after onset of papillary muscle rupture, emergency operation was performed. At operation, posterior papillary muscle was found to be totally ruptured. Coronary artery revascularization and mitral valve replacement were performed. Postoperative course was uneventful, with 4 days of IABP and 5 days of ventilatory support. She was discharged on the twentieth postoperative day in NYHA class I. Reports of successful emergency operation for total papillary muscle rupture following acute myocardial infarction are rare. Early diagnosis and surgical treatment are mandatory to save this group of patients. PMID- 9847581 TI - [A case of tuberculous granulomatous mediastinitis and constrictive pericarditis simulating malignant mediastinal tumor]. AB - A case is 40-year-old man. He presented anterior chest pain. Pericardial effusion was pointed out and a tuberculin skin test was positive. Tuberculous pericarditis was highly suspected, so INH and RFP were medicated. After 6-month medication pericardial effusion decreased, but right pleural effusion appeared on chest X ray. Chest CT revealed a thickening of pericardium extend to anterior mediastinal mass. Echocardiogram revealed a pressure gradient in right ventricle, which was compressed by the thickened pericardium. We underwent median sternotomy in order to rule out neoplastic diseases. Intraoperative pathologic diagnosis was granulomatous mediastinitis and pericarditis, so we resected granuloma as much as possible to decompress the heart. Although Mycobacterium tuberculosis was not found in the resected granuloma, it was most probable pathogen. He received additional antituberculous chemotherapy for 6 months. PMID- 9847582 TI - [One stage operation for aneurysm of the diverticulum of the ductus arteriosus and coronary artery bypass grafting]. AB - Aneurysm of the diverticulum of the ductus arteriosus in the adult is rare. One stage operation for aneurysm of the diverticulum of the ductus arteriosis and coronary artery bypass grafting (CABG) is reported. A 61-year-old man was admitted for diagnosis of thoracic aneurysm on chest X-ray and CT. Chest CT scan showed an aneurysm above the left main pulmonary artery. An aortography showed the left vertebral artery originated directly from the aortic arch and a saccular aneurysm arising from the aortic isthmus and lesser curvature of the aortic arch. Coronary arteriography showed 75% stenosis at the right coronary artery (seg. #1) and 75% stenosis at the left anterior descending artery. Operation was performed through a median sternotomy. The aneurysm of 6 to 3 cm was located between the aortic isthmus and left pulmonary artery. Ascending aorta and right atrium were used to institute cardiopulmonary bypass (CPB). CABG (LITA to #7, SVG to #4 PD) was performed. Arterial cannulation was then switched to the left femoral artery. The proximal aorta was cross-clamped between the left vertebral artery and the left subclavian artery under the partial CPB, and the distal aorta was occluded with a occulusive balloon catheter via the right femoral artery. The selective left axillar artery cannulation was performed to perfuse LITA. The aneurysm was resected and closed with a patch. His post-operative course was uneventful. PMID- 9847583 TI - [A case of racemose hemangioma of the right bronchial artery and intercostal to pulmonary arterial anastomosis]. AB - We reported a case of racemose hemangioma of the bronchial artery and intercostal to pulmonary arterial anastomosis. A 67-year-old woman was admitted because of repeated hemoptysis. Bronchoscopic examination revealed a torous lesion of the right B7 bronchus. Intercostal angiography demonstrated communications between right dilated, meandered intercostal arteries and right pulmonary artery. Bronchial angiography showed dilatation and convolution of the right bronchial artery. Angiographic embolization of the right bronchial artery and the right intercostal artery was underwent. There was no recurrence of hemoptysis one year after the embolization procedure. We think that angiographic embolization is an effective method of treatment of hemoptysis due to racemose hemangioma of the bronchial artery and intercostal to pulmonary arterial anastomosis. PMID- 9847584 TI - [A case report of incomplete endocardial cushion defect with mitral stenosis in an elderly patient]. AB - The following paper describes a mitral valve replacement (SJM 27 mm), the patch closure (EPTFE) of an ostium primum atrial septal defect and tricuspid annuloplasty (De Vega's method) in a 64-year-old female patient with an incomplete endocardial cushion defect and mitral stenosis. Surgery revealed thickened, mitral valve leaflets and the presence of a cleft, findings similar to those observed in case of rheumatic degeneration. Investigation of patient hemodynamics confirmed a diagnosis of Lutembacher syndrome and a lower with left ventricle volume. After surgery, the volume of left ventricle increased and the patients clinical course was uneventful. PMID- 9847585 TI - [A case of successful resection of pulmonary arteriovenous fistula with a high shunt ratio]. AB - We report on a 25-year-old cyanotic man who was diagnosed as having a pulmonary arteriovenous fistula. His chief complaint had been shortness of breath since childhood. Polycythemia (Hb 21.4 g/dl) was detected during a health checkup at his company. A chest X-ray showed an abnormal mass in the left lung. Blood gas analysis showed severe hypoxia with PaO2 of 38.6 mmHg at room air. Angio-CT showed a large aneurysmal lesion at S6 of the left lung with a large feeding artery and vein. Oxygen saturation was 75.2% in the radial artery and 62.5% in the right atrium. The right-to-left shunt ratio was therefore calculated as 62%. The aneurysmal lesion was resected by segmentectomy of the left S6 following division of A6 and V6. After a successful operation, the patient no longer had shortness of breath or cyanosis and blood gas analysis showed PaO2 as 84.3 mmHg at room air. Pulmonary angiography showed no residual shunt lesion. PMID- 9847586 TI - [A combination of reoperation for pseudoaneurysm following the Cabrol procedure and total aortic arch replacement in a patient with Marfan syndrome--a case with an aberrant right subclavian artery]. AB - A 44-year-old male with Marfan's syndrome had undergone an initial operation for DeBakey type I acute aortic dissection with annulo-aortic ectasia. He had undergone replacement of the ascending aorta and aortic valve with a composite graft and reconstruction of the coronary artery by the Cabrol procedure. At 5 years after the initial surgery he experienced chest pain and was subsequently examined. Computed tomography revealed a pseudoaneurysm in the ascending aorta and the residual aortic dissection. The maximum diameter of the pseudoaneurysm was 85 mm and the maximum diameter of the aortic arch was 55 mm. The aortic arch was associated with an aberrant right subclavian artery. Angiography revealed that the pseudoaneurysm was caused by leakage at the coronary ostium-graft anastomoses. We repaired the anastomoses and performed total aortic arch replacement with reconstruction of four arch branches. The postoperative course was uneventful without any complications. We report this case because there have been few reports regarding arch replacement in cases with an aberrant right subclavian artery. PMID- 9847587 TI - [Prediction of spinal cord ischemia with a retrievable stent graft on endovascular treatment for a case of thoracic aortic aneurysm]. AB - Multiple aortic aneurysms in Behcet's disease were repaired with transluminaly placed endovascular stent grafts. Before deploying the stent graft device for permanent implantation for the saccular aneurysm located in the descending thoracic aorta, from which feeding arteries for the spinal cord possibly branched, a retrievable stent graft was inserted and evoked spinal cord potential (ESP) were monitored in order to predict spinal cord ischemia. The original retrievable stent graft, constructed of a self-expandable Z-shaped stainless steel stent covered with e-PTFE, can be easily withdrawn into a 18 Fr. sheath after deployment. Blood flow into intercostal arteries branching from that part of the descending aorta where the permanent stent graft is planned to be implanted, is intercepted by the retrievable stent graft. A change of ESP during the temporary implantation of the device indicates that spinal cord ischemia would be caused by permanent implantation of the stent graft. In this case, no change of ESP was observed and the patient showed no postoperative paraplegia. The retrievable stent graft was useful for prediction of spinal cord ischemia before endoluminal stent graft repair of the descending aortic aneurysm. However, the device is not flexible enough to fit a severely tortuous aorta, therefore we are obliged to select patients to some extent. Further improvement of the device is required to make prediction of spinal cord ischemia with the retrievable stent graft possible in all cases. PMID- 9847588 TI - [Two cases of minimally invasive reoperative coronary artery bypass]. AB - Reoperative coronary artery bypass grafting (CABG) are still associated with higher mortality than primary CABG. This is due in part to the potential for cardiac and patent graft injury during their dissection and the reopening of the sternum. Therefore, in two patients with recurrent angina attributable to occlusion of the old vein graft to the LAD, we performed reoperative CABG by the minimally invasive direct coronary artery bypass (MIDCAB) procedures. The left internal thoracic artery was anastomosed to the LAD through small anterolateral thoracotomy without cardiopulmonary bypass. Both patients recovered fast and underwent postoperative angiogram, showing the new grafts widely patent. About two weeks later, both discharged in the conditions of nearly normal activities. The reoperative MIDCAB grafting might be expected to be as safe and promising as the primary one. PMID- 9847589 TI - [A case of cardiac lipoma in the ventricular septum]. AB - Lipoma in the ventricular septum is very rare. Our review of the English literature revealed that our case is the sixth of removal of lipoma in the ventricular septum. A 60-year-old male was admitted because a mass in the ventricular septum was found incidentally in abdominal CT taken following type B hepatitis. CT scanning of the heart showed 2 cm diameter of tumor in the ventricular septum. The tumor had very low radiodensity, so it was thought to be identical to fat tissue. The tumor was more clearly visualized by MR imaging and the signal intensity was high on the T1-weighted image. The tumor was suspected to be lipoma. The intraoperative histological diagnosis showed the tumor was lipoma. The tumor adhered strictly on the myocardium of the septum, and it was located near the left anterior descending coronary artery. It could not completely resected, in these reasons. Postoperative course was uneventful, and echocardiogram taken 9 months after the operation showed no evidence of enlargement of the residual tumor. It is necessary to follow-up rigidly for the potential of enlargement of the residual tumor. PMID- 9847590 TI - [Reoperation for diffuse supravalvular aortic stenosis with Williams syndrome- extended patch aortoplasty and extra-anatomic bypass from the ascending aorta to the descending aorta in a median sternotomy]. AB - A case of diffuse supravalvular aortic stenosis (SVAS) with Williams syndrome is reported. In this case of severe diffuse SVAS, we performed the diamond-patch aortoplasty in a child. However he has been suffering from residual SVAS. At 9 years old, the myocardial injury was noted by myocardial scintigraphy. Preoperative cardiac catheterization and angiography revealed the hypoplastic ascending aorta and arch with a pressure gradient of 89 mmHg at the distal site from the left subclavian artery. Through only a median stenotomy, an extended patch aortoplasty between the valsalva sinus and distal arch was performed and an extraanatomic bypass from the ascending aorta to the descending aorta was employed using a 10 mm tube graft. We realize this technique is available because this method can relieve the left ventriculus of the pressure load and operate via only median sternotomy. PMID- 9847591 TI - [A case of the ascending aorta and aortic arch replacement with thrombo-occlusion of distal arch and descending aorta surround the modified elephant trunk graft]. AB - We report a 65-year-old female who had a extensive thoracic aneurysm from ascending aorta to descending thoracic aorta. The patient underwent a graft replacement of ascending aorta and aortic arch using modified elephant trunk method. The surgery was carried out through median sternotomy with profound hypothermia and selective cerebral perfusion. Postoperatively, the patient was recovered without any complications except bronchial asthma. Postoperative chest computed tomography showed that the surrounding space of the elephant trunk vascular graft inserted into distal arch and descending aneurysm was mostly occupied with thrombus. Therefore, we considered that the second operation on the descending aorta is not necessary at this point and careful attention to the size and shape of the descending aneurysm should be paid. PMID- 9847593 TI - [The utilities and outlooks of MR cholangiography (MRCP) as a non contrast and noninvasive technique]. AB - Endoscopic retrograde cholangiopancreatography (ERCP) is regarded as the diagnostic procedure of decision for abnormalities of the biliary and pancreatic ducts. However, ERCP is unsuccessful in 3-10% cases and is also associated with a 1-5% risk of producer-induced pancreatitis and other complications. MR cholangiography (MRCP) has recently developed as a noninvasive examination for evaluation of pancreaticobiliary diseases. Heavily T2-weighted fat-suppressed sequences are used to obtain these images. On MRCP, static fluid, such as bile or pancreatic juice represents hyperintense without injection of contrast medium. Therefore, MRCP could be examined in almost all patients including as infant or the patients of post reconstruction of gastrointestinal tract without any complication. In this paper, I described the several utilities and outlooks of MRCP as a noninvasive technique through the representative clinical applications. PMID- 9847594 TI - [Basic principles and historical consideration of MR cholangiopancreatography]. AB - Basic principle of MR cholangiopancreatography (MRCP) is heavily T2-weighted imaging (hydrography) to use bile and pancreatic juice as "natural contrast medium". Firstly developed sequence for MRCP was a CE-FAST sequence, which employed time-reversed FID signal. The current most popular sequences for MRCP are single-shot fast spin-echo sequences, which are divided into three types (2D single slice, 2D mutiple slice and 3D methods). The advantage of 2D single slice method is conveniently obtained projection imaging within a few seconds of examination time. Both 2D multiple slice and 3D methods consists of a MIP image and a series of source images. The MIP image creates global images of pancreatico biliary system. The source images provide detailed evaluation of various anatomical structures and abnormalities. By using these sequences properly, MRCP can yield valuable informations of pancreatico-biliary diseases non-invasively. PMID- 9847595 TI - [Maximum intensity projection (MIP) and multiplanar reformation (MPR) for post processing cholangiopancreatographic data set--clinical application and pitfalls]. AB - Maximum intensity projection (MIP) and multiplanar reformation (MPR) are the most frequently used algorithms for MR cholangiopancreatography (MRCP). The MIP allows three dimensional overview of the pancreatic and biliary system. Because of its resemblance to ERCP images, MIP reconstruction is widely accepted by clinicians. In spite of its usefulness, MIP may be misleading without a proper reference to source images or a guidance of MPR. Opacification defects that reflect intra ductal or intra-cystic pathologies are notably erased through the process of MIP reconstruction. Diagnosis based only on MIP images is therefore not clinically feasible. A combined use either of multisection images, or at least source images, is essential. The MPR on the other hand, enables an investigation of the details of the intra-ductal or intra-cystic pathologies. Detailing with MPR and surveying with MIP work together in interpreting MRCP data set. PMID- 9847596 TI - [Clinical evaluation of MRCP]. AB - Recently, MRCP can be obtained with good spacial resolution within a few seconds using half fourier fast spin echo technique. From July 1995 to August 1997, 1000 patients suspected of having pancreatobiliary diseases were examined with MRCP. MRCP was performed with 1.5 T scanner using Fast Asymmetric Spin Echo sequence (FASE). Satisfactory images of the main pancreatic duct were obtained in 98%, of Santorini's duct in 90%, and of uncinate process branch in 83%. Pancreas divism was accurately diagnosed. In the patients with pancreatic ductal adenocarcinomas, MRCP depicted stenosis and proximal dilatation of the main pancreatic duct in 89%, and in the remaining patients no abnormalities were seen in the main pancreatic ducts because tumors were limited to side branches or Santorini's duct or distal end of the tail of the pancreas. Diagnosis of small pancreatic carcinomas (smallest lesion measured 10 mm in diameter) were feasible. In the patients with intraductal papillary neoplasms, dilatation of the main pancreatic duct and cystic dilatation of side branches were depicted, and polypoid lesions were detectable with source images. In the patients with serous cystadenomas, accumulation of the microcysts were visualized. In the patients with chronic pancreatitis, dilatation and stenosis of the main pancreatic duct, as well as side branch dilatation was depicted despite overestimation of the extent of the stenosis. Stones in the main pancreatic duct were well visualized as intraductal filling defects. In conclusion, MRCP is an effective imaging technique in the diagnosis of various pancreatic diseases. PMID- 9847597 TI - [PSIF/CE-FAST (fast gradient echo) and TSE (turbo spin echo)]. AB - Imaging techniques of PSIF/CE-FAST (fast gradient echo) and TSE (turbo spin echo) are described. PSIF/CE-FAST is a mirror imaged FISP (fast imaging with steady state precession) technique which is applied to MR cholangiopancreatography (MRCP) for the first time. However, it has been replaced by TSE technique because of poor resolution and motion artifacts. TSE is a fast imaging technique using multiecho method, which enables us to obtain high resolution MRCP with heavily T2 weighted images. MRCP using TSE shows high signal to noise ratio with minimal motion artifacts. Recently, breath-hold single shot fast imaging techniques within a few seconds or respiratory triggered TSE are applied to MRCP and shows higher resolution for pancreatobiliary ducts system. PMID- 9847598 TI - [FASE (fast advanced spin echo)]. AB - RF-refocused single or multi-shot EPI provides high-contrast 2D or 3D T2-weighted images in a very short scan time. Applications include MR cholangiopancreatography, MR myelography, imaging detailed structures of the internal auditory canal and in situations in which very fast T2 imaging is required. FASE offers both 2D and 3D techniques. 2D FASE technique permits high resolution images of 384 matrix or more to be obtained at 2 to 3 seconds per image. It is easy to perform and suitable for screening. 3D FASE permits acquisition of isotropic voxels, allowing high-resolution viewing from any desired direction by post-processing (MIP and/or MPR). This technique is best suited to detailed examinations in which multiple projection angles will be reconstructed or high-resolution diagnosis of source images. PMID- 9847599 TI - [Single shot fast spin echo (SSFSE)]. AB - Single Shot Scan methods, which acquire all the necessary data for reconstruction with one excitation, are very useful methods to minimize motion artifacts. Single Shot Scan can be categorized in 2 groups, one is EPI method using gradient by Echo data acquisition, and another is SSFSE method using RF. SSFSE method is Single Shot FSE method using 0.5 NEX. SSFSE produces images with less motion artifacts and heavy PD and T2 images with high contrast. New RF reduces Echo Space and minimizes image quality degradation caused by Single Shot. SSFSE is useful for long T2 like MRCP. Long TE mode can be used with very long ETE even with 0.5 NEX. New applications development for Single Shot using RF is awaited in the future. PMID- 9847600 TI - [Characteristics and detectability of RARE technique]. AB - RARE is a kind of fast spin echo technique with large number of echo train and long TE. Because of long TE, signal from parenchyma is vanished and only liquid with long T2, such as bile, pancreatic juice, is detected. Compared to HASTE, signal from vessels like portal or hepatic vein is vanished either. Acquisition time of RARE is extremely short because it is single shot technique. So, on MRCP examination, RARE is good method for easily understanding of whole structure of biliary system because it is usually used as a projection image with thick slice. On the other hand, one should pay attention for shorter T2 bile and pancreatic juice. If their T2 is short, biliary system may not be shown with RARE. PMID- 9847601 TI - [Optimal scan parameters of MRCP using HASTE]. AB - For improving the image of fine structures on MRCP, optimal scan parameters were assessed by water-filled phantom using HASTE sequence. Spatial resolution and Contrast-to-Noise Ratio (CNR) were measured at various FOVs and slice thicknesses with a body array coil. High spatial resolution was provided at less than 25 cm in FOV, and narrow phantom under 1 mm in diameter was obscured at more than 30 cm in FOV. Highest CNR was provided at 3 mm slice thickness on a 1.5 T unit, and at 4 mm slice thickness on a 1.0 T unit. Narrow phantom under 1 mm in diameter was depicted clearly on a 1.5 T unit, but irregularly on a 1.0 T unit. Taking required spatial resolution and CNR into consideration, we should determine optimal FOV and slice thickness for the assessment of anatomic details on MRCP. PMID- 9847602 TI - [Optimization of pulse sequences for MR cholangiopancreatography with FASE]. AB - Various types of pulse sequences for MR cholangiopancreatography (MRCP) have been developed in the past ten years. FASE (fast advanced spin echo) is one of the single-shot fast spin-echo sequences developed by Toshiba. By using this sequence, 2D single slice, 2D multiple slice and 3D methods can be provided. We routinely employ 2D single slice and 3D methods. The merit of 2D single slice method is conveniently obtained projection imaging within short data acquisition time. On the other hand, 3D method can yield detailed evaluation of various anatomical structures and abnormalities by using thin-slice source images. O2 inhalation study is especially recommended in case of 3D method. With the proper knowledge of sequence characteristics, MRCP using FASE can provide valuable informations of pancreatico-biliary diseases. PMID- 9847603 TI - [Breath-hold MR cholangiopancreatography with 2D and 3D-FASE sequence at 0.5 tesla: evaluation based on measurements of signal intensity ratio and contrast-to noise ratio]. AB - Single-slice 2D-FASE (more than 2 cm) and 3D-FASE images proved to be reliable techniques by which to create high-resolution MRCP images at 0.5 T. In particular, 3D-FASE sequence can provided MIP and its thin source images with sufficient SIR and CNR of the images, which are useful to evaluate detailed structures. Using 3D-FASE sequence, even the non-dilated pancreatobiliary system can frequently be demonstrated because each source image retains satisfactory SIR and CNR of the images due to 3D acquisition and is not affected by motion artifacts of any kind. The most important problem for 3D-FASE sequence is that view-to view amplitude modulation errors may persist, though gradient moment errors can be reduced because of no actual motion during acquisition, resulting in degraded the MIP images due to the different positions of the source images in a few patients with inconstant respiration. PMID- 9847604 TI - [MR-cholangiopancreatography using three-dimensional Fourier transfer fast asymmetric spin-echo method (3DFT-FASE)--clinical evaluation]. AB - MR-cholangiopancreatography (MRCP) is a new and non-invasive method. With the three-dimensional Fourier transform fast asymmetric spin-echo (3DFT-FASE) method that allows data collection as volume and three-dimensional expression, a higher spatial resolution can be obtained in the direction of both slab thickness and in plane. In this method, analysis by multi planar reconstruction (MPR) and observation from various angle by maximum intensity projection (MIP) are available. Although further studies are required to shorten the imaging time to stop breathing only once, to use the respiratory gating or navigator echo, clinically useful diagnosis are possible utilizing by the advantages of the 3DFT method. PMID- 9847605 TI - [Demonstration of normal bile duct and pancreatic duct with MR cholangiopancreatography]. AB - Demonstration of normal bile duct and pancreatic duct with MR cholangiopancreatography was assessed in 78 patients including 19 cases of not impacted gallstone. Breath hold 2D-MRCP and intermittent breath hold 3D-MRCP was performed with half-Fourier fast spin echo technique using 1.5 T imager and surface coil. Diagnostic depiction rates of main pancreatic duct were 99% in the head, 97% in the body, and 91% in the tail, and of extrahepatic bile duct was 100%. Visualization of accessory pancreatic duct was 42%, but of side branch of pancreatic duct in the body to tail was poor. Anatomic variants of the biliary tree were seen in 15 cases. The reason of poor demonstration was the poor breath hold. In the majority of patients, MRCP can provide the accurate information of pancreaticobiliary tract without dilatation. PMID- 9847606 TI - [Usefulness of T2-weighted images using single shot fast spin echo (SSFSE) pulse sequence for the evaluation of pancreatobiliary diseases: comparison with MRCP using SSFSE]. AB - Single shot fast spin echo (SSFSE) pulse sequence provides us with a thick single slice MR cholangiopancreatography (MRCP) as well as thin multi-slice MRCP, and T2 weighted images are also obtained with SSFSE sequences. In comparison with conventional spin-echo and fast spin-echo sequences, T2-weighted image using SSFSE sequence has the advantages of the abscence of motion artifacts and the extremely short acquisition times. MRCP using SSFSE sequence is useful in the conspicuity of bile duct, pancreatic duct, and cystic lesions. However, T2 weighted image using SSFSE is superior to MRCP not only for the differentiation between solid and cystic lesions but also for the visualization of contour of different organs and lesions. PMID- 9847607 TI - [MR cholangiopancreatography with single-shot fast-spin-echo sequences]. AB - Imaging quality of MR cholangiopancreatography (MRCP) has recently made a great advance, and MRCP plays an important part in the diagnosis of pancreticobiliary diseases. To obtaining excellent-quality MRCP images, three conditions are required: fluid-to-background contrast, high spatial resolution, and suppression of respiratory motion artifacts. Respiratory motion artifacts, whose suppression is especially important for MRCP, can be controlled by the use of breath-holding, signal averaging, or respiratory triggering. These images are implemented in either single-slice technique or multi-slice technique. We have performed MRCP prior to endoscopic retrograde cholangiopancreatography (ERCP) in more than 100 patients. MRCP images were obtained as maximum-intensity-projection (MIP) reconstruction images and thick-slice projection images by using single-shot fast spin-echo sequence, and MIP reconstruction images by using respiratory-triggered fast-spin-echo sequence. We reviewed MRCP imaging with single-shot fast-spin-echo sequences. PMID- 9847608 TI - [Comparative studies of breath-hold magnetic resonance cholangiopancreatography (MRCP) between different two sequences and between 1.0T and 1.5T units]. AB - In this article, a comparative study in the diagnostic ability of a breath-hold MRCP between thin slice half-Fourier acquisition single-shot turbo spin-echo (HASTE) and thick slice rapid acquisition with relaxation enhancement (RARE), and between 1.5T and 1.0T units was made. Although thin slice HASTE sequence might be more excellent in the diagnostic ability than thick slice RARE, the ability of depiction of pancreaticobiliary system on MRCP was not statistically different between these two sequences and between 1.0T and 1.5T units. Thick slice RARE sequence was inferior to thin slice HASTE in the depiction of gallbladder stone. However, thick slice RARE sequence was useful for MRCP because a projection image of pancreaticobiliary system could be easily obtained without postprocessing and without misregistration. Thick slice RARE sequence should be used in combination with thin slice HASTE sequence for the diagnosis of pancreaticobiliary system. PMID- 9847609 TI - [Elimination of artifacts in MRCP: technical consideration]. AB - Artifacts of MR cholangiopancreatography (MRCP) include fluid-filled gastrointestinal tracts, respiratory motion artifacts, spasm of Oddi's sphincter, vascular compression of bile ducts, overlapping of various anatomical structures and bright signal of surrounding fatty tissue. In this article, various technical considerations to eliminate these artifacts were described for the better imaging analysis of MRCP. The use of high-concentration ferric ammonium citrate (Ferriseltz) is recommended to reduce bright signal of fluid-filled gastrointestinal tracts. In case of long breathhold sequences, O2-inhalation study is useful to eliminate respiratory motion artifacts. Careful attention should be paid to the spasm of Oddi's sphincter and the vascular compression of bile ducts to avoid erroneous interpretation of MRCP findings. PMID- 9847610 TI - [MR cholangiopancreatography using respiratory triggered half-Fourier RARE technique]. AB - We evaluate the usefulness of MRCP using respiratory triggered HASTE technique in 68 patients. Eleven subsequent slices were obtained for MRCP. To determine the optimal trigger rate, MRCP with various trigger rates were compared in 25 patients. Respiratory triggered MRCP and breath hold MRCP were compared in 68 patients. Respiratory triggered MRCP was also performed in 3 infants. The optimal trigger rate was 10%. In 17 patients who could not stop their breath for 10 seconds, respiratory triggered MRCP were better than breath hold MRCP in 14 (82%) for the source images and in 17 (100%) for the MIP images. Respiratory triggered MRCP could depict cystic duct and main pancreatic duct even in 14 month infant. MRCP with high spatial resolution could be obtained with respiratory triggered HASTE technique in critically ill, or non-cooperative patients. PMID- 9847611 TI - [Clinical efficacy of suppression and visualization of the duodenal fluid in MR cholangiopancreatography using ferric ammonium citrate]. AB - The high signal intensity from the intestinal fluid usually deteriorates the projection images of MR cholangiopancreatography (MRCP) because it superimposes on the biliary tract. FerriSeltz (Otsuka pharmaceutical, Tokushima, Japan; a ferric ammonium citrate-based positive oral contrast agent) solution in a twelve fold higher concentration has a significant T2 shortening effect and could be used as a negative contrast agent suppressing the signal from intestinal fluid. Recently, we developed a new method named as "Dual Contrast MRCP" with intermediately concentrated FerriSeltz solution. This method provides MRCP both with and without duodenal image depending on TE length and is worth while screening of duodenal lesion even in a case with a small mount of duodenal fluid. PMID- 9847612 TI - [Clinical usefulness of fat suppression technique for MR cholangiopancreatography (MRCP)]. AB - In order to obtain high quality of MRCP image, it is important to reduce the background signal intensity and increase the contrast of the pancreatobiliary ducts relative to surrounding fat tissue. The combination of long effective-TE and fat suppression technique including the short-TI inversion recovery and chemical-selective fat suppression enables to suppress the background signal intensity enough to obtain high quality MR cholangiopancreatogram. However, susceptibility artifacts from the metal, gastroduodenal gas, and vascular pulsation can be augmented by using the chemical-selective fat suppression technique, which may result in signal loss of the pancreatobiliary ducts. This potential diagnostic pitfalls can be avoided by interpreting the coronal source images obtained with long effective-TE and without fat suppression technique. PMID- 9847613 TI - [Evaluation of MRCP compared to ERCP in the diagnosis of biliary and pancreatic duct]. AB - Recently, Magnetic Resonance Cholangiopancreatography (MRCP) is developed as a noninvasive diagnostic modality in the diagnosis of biliary and pancreatic tract. Using Endoscopic Retrograde Cholangiopancreatography (ERCP) as the gold standard, we evaluated the diagnostic quality of MRCP and direct cholangiography. Fifty-six patients (9 cases of cholangiocarcinoma, 5 of gallbladder carcinoma, 1 of gallbladder carcinoma and anomalous arrangement of pancreaticobiliary ductal system, 4 of cholecystlithiasis, 3 of papillary carcinoma, 1 of adenomyomatosis of the gallbladder, 1 of primary sclerosing cholangitis, 1 of hepatolithiasis and postoperative bile duct stricture, 4 of mucin producing pancreatic tumor, 13 of pancreatic carcinoma, 1 of chronic pancreatitis and pancreas divisum, 9 of chronic pancreatitis, and 1 of chronic pancreatitis and biliary stricture) are studied prospectively with MRCP and direct cholangiography (included ERCP and percutaneous transhepatic cholangiography). The image of MRCP accorded with direct cholangiography in twenty-two of 27 patients with biliary tract disease, and in sixteen of 29 patients with pancreatic disease. The different diagnosis is observed in ten of 56 patients. In cases of not visualized gallbladder, pancreatic cyst without communicated to the pancreatic duct, and pre-stenotic dilatation of biliary and pancreatic duct, the image of MRCP was better than that of ERCP. However, the image of MRCP for the diagnosis of either benign or malignant strictures, mucin producing pancreatic tumor, and a branch of pancreatic duct in patients with pancreatic carcinoma were not suitable for evaluation than that of ERCP. In our study, ERCP is superior to MRCP due to the important information for diagnosis such as mentioned above. Therefore we advocate using ERCP as the first diagnostic tool in the diagnosis of biliary and pancreatic duct. PMID- 9847615 TI - [Value of MR cholangiography in the diagnosis of pancreatic diseases compared with endoscopic retrograde cholangiopancreatography]. AB - PURPOSE: To compare magnetic resonance cholangiography (MRCP) with endoscopic retrograde cholangiography (ERCP) in the diagnosis of pancreatic diseases. METHODS: The authors retrospectively reviewed MRCP and ERCP images obtained in 56 patients with pancreatic diseases and 192 controls. Detectability of the pancreatic duct and its branches and diagnostic usefullness were evaluated. RESULTS: MRCP was successful in almost all the patients without ascitic fluid or ileus. Findings of the main pancreatic ducts (i.e., dilation, stenosis and obstruction) were similarly demonstrated by ERCP and MRCP. The degree of ductal narrowing tends to be overestimated with MRCP. CONCLUSION: MRCP appears to be less invasive than ERCP. MRCP could provide sufficient information and play a complementary role in the diagnosis of pancreatic diseases. PMID- 9847614 TI - [The usefulness of MRCP in the initial diagnosis of the biliary and pancreatic diseases compared with ERCP]. AB - The purpose of this study was to determine the effectiveness and the limitation of the MR cholangiopancreatography (MRCP) in the initial diagnosis for the biliary and pancreatic diseases compared with the image of Endoscopic retrograde cholangiopancreatography (ERCP). MRCP showed higher performance toward the discovery of the choledocholithiasis of a diameter 4 mm and more. Furthermore, MRCP reflected a malignant tumor at the constrictive part and the dilated proximal part of the biliary and pancreatic duct in 23 example all. An advantage of ERCP was the ability to carry out biopsies and therapeutic procedures, such as biliary drainage and sphincterotomy, while MRCP was an important diagnostic modality in the work-up of the biliary and pancreatic diseases and could help in planning treatment. MRCP would become the examination method first used, and it would be popularized all the more from now on. PMID- 9847616 TI - [MR cholangiopancreatography of pancreas divism and anomalous arrangement of the pancreaticobiliary ducts]. AB - The purpose of this study was to evaluate the usefulness of MR cholangiopancreatography (MRCP) for pancreas divism and anomalous arrangement of pancreaticobiliary ducts (APBD). In 214 patients, MRCP was performed. In 204 patients (94%), pancreaticobiliary ductal junction could be recognized. Pancreas divism was observed in 13 of 204 patients (6%), and APBD was observed in 10 of 204 patients (5%). Dynamic MRCP after secretin stimulation was useful for diagnosing pancreas divism and APBD. PMID- 9847617 TI - [Chronic pancreatitis, acute pancreatitis]. AB - MRCP has been recognized as a safe and noninvasive diagnostic method. In the present study we evaluated the usefulness of MRCP in diagnosis of chronic and acute pancreatitis. Two-dimensional fast asymmetric spin-echo (FASE) MRCP was performed in 40 patients with chronic pancreatitis and 13 with acute pancreatitis. In 29 patients (72.5%) with chronic pancreatitis and 9 (66.7%) with acute pancreatitis, main pancreatic duct (MPD) was visualized entirely. MRCP could demonstrate the characteristic findings of chronic pancreatitis such as dilatation and irregularity of MPD in most cases. In acute pancreatitis, MRCP indicated that MPD was normal in diameter, but irregular in configuration compared with that of the control group. MRCP may facilitate the diagnosis of chronic and acute pancreatitis. PMID- 9847618 TI - [Cystic lesions of the pancreas]. AB - The number of the literature and classification of the cystic pancreatic diseases is increasing recently. We describe MR cholangiopancreatography (MRCP) findings of the cystic pancreatic diseases according to the clinical oriented classification. Intraductal papillary tumor, mucinous cystadenoma and serous cystadenoma showed characteristic MRCP findings. However small non-neoplastic true cysts are difficult to differentiate from cystic tumors even by MRCP. PMID- 9847619 TI - [MR cholangiopancreatography (MRCP) for the mucin producing pancreatic tumor (MPT)]. AB - MRCP was performed for 17 patients with MPT (main duct type; 7 and branch duct type; 10) using HASTE and single shot RARE with a body phased array coil on a 1.5 T unit. The findings of MRCP were compared with those of endoscopic retrograde cholangiopancreatography (ERCP). In all cases, MRCP completely disclosed all dilated ducts and cysts, some of which were not seen by ERCP due to mucinous accumulation. In five of seven patients with main duct type, an excrescent nodle was more clearly visualized by ERCP than by MRCP. Mucinous accumulation was not confirmed by MRCP in all cases except one case, in which the dilated branch duct was revealed as lower intensity compared with the main duct using single shot RARE. Therefore MRCP and ERCP are complementary methods in diagnosis for the mucin producing pancreatic tumor. PMID- 9847620 TI - [Clinical efficacy of MRCP for diagnosis of patients with pancreatic neoplasm]. AB - We examined the usefulness of MRCP in pancreatic cancer compared with ERCP. Moreover, it examined in the same way in focal pancreatitis, too. We compare three findings of 1) main pancreatic duct, 2) branches of the stricture part, 3) side of tail main pancreatic duct about ERCP and MRCP. As for the branches of the stricture part, MRCP was poor compared with ERCP but as for the property of the side of the tail main pancreatic duct, MRCP excelled. The study of mass forming chronic pancreatitis was a similar result and was not the one, which excels ERCP in case of qualitative diagnosis. However, we thought that it was not invasively, suitable behind the digestive operation and digestive passage fault, in case of elapse observation and moreover that MRCP is suitable for pick up the pancreatic cancer in our clinic. PMID- 9847621 TI - [Anatomic variants of the cystic duct and cysticohepatic junction: diagnosis with MR cholangiography]. AB - The cystic duct are variable in length, course and site of termination. A knowledge of the variable anatomy of the cystic duct and cysticohepatic junction is important in biliary surgery, because failure to recognize anatomic variations may result in a significant ductal injury. Magnetic resonance cholangiography (MRC) is a recently developed technique that demonstrates the biliary tree noninvasively and without injection of contrast material. Anatomic variants of the cystic duct and cysticohepatic junction that may increase the risk of bile duct injury in biliary surgery are frequently identified with MRC. MRC will be a noninvasive and a useful technique in the diagnosis of anatomic variants of the cystic duct and cysticohepatic junction. PMID- 9847622 TI - [Diagnostic usefulness of magnetic resonance cholangiopancreatography (MRCP) in comparison with retrograde cholangiopancreatography (ERCP) for cholelithiasis]. AB - PURPOSE: To elucidate MRCP diagnostic usefulness compared to ERCP. PATIENTS, METHODS: 29 MRCP performed patients diagnosed by ERCP were studied. RESULTS: MRCP showed 92%, 67% sensitivity and 73%, 100% specificity in choledocholithiasis (CBD) and cholecystolithiasis (GB), respectively. MRCP revealed 4.0 false positive and 1.5 false negative in CBD and GB, respectively. MRCP could detect 22 stones out of 33 in CBD and 13 out of 42 in GB. As for the maximal diameter, MRCP depicted 31% for less than 5 mm and 100% for over 6 mm in CBD. In GB, 7% for less than 5 mm, 83% for 6-10 mm and 100% for over 11 mm. CONCLUSION: Although the depection of tiny stone is limited, MRCP is a useful diagnostic tool for cholelithiasis, especially choledocholithiasis. PMID- 9847623 TI - [Common bile ductal carcinoma, hilar cholangiocarcinoma: evaluation with MR cholangiography]. AB - Recently, imaging of bile ductal carcinoma has been improved by new techniques such as MR cholangiography. Cancer invasion along the bile duct has been demonstrated by MR cholangiography. MR cholangiography will be useful for obtaining more detailed information in the diagnosis and staging of common bile ductal carcinoma, hilar cholangiocarcinoma. PMID- 9847624 TI - [MR cholangiopancreatography of Mirizzi syndrome and Lemmel syndrome]. AB - The purpose of this study is to evaluate the usefulness of MR cholangiopancreatography (MRCP) in diagnosing Mirizzi syndrome and Lemmel syndrome. MRCP could show compressed common hepatic duct by enlarged gallbladder in Mirizzi syndrome. Periampulary duodenal diverticulum and compressed lower bile duct can be recognized clearly by MRCP after secretin stimulation. So MRCP should apply for diagnosing of Mirizzi syndrome or Lemmel syndrome. PMID- 9847625 TI - [Findings of MR and MR cholangiopancreatography in acute cholecystitis]. AB - We retrospectively reviewed magnetic resonance cholangiopancreatography (MRCP) of 25 patients with acute cholecystitis based on clinical, sonographic and surgical findings. Intramural high signal intensity on MRCP was demonstrated in 22 of the 25 patients (88%), and pericholedochal high signal intensity was observed in 6 of the 25 patients (24%). Pericholecystic or perihepatic fluid was demonstrated in 6 of the 25 patients (24%). Gallbladder stones were identified in all 21 patients (100%) by sonography, in 19 of the 21 (90%) by MRCP and in 11 of 18 patients by CT (CT was not performed in other 3 patients). Common bile duct calculi were detected in all 6 patients (100%) by MRCP, in 2 of the 6 (33%) by sonography, and in 5 of the 6 (50%) by CT with confirmation of surgical finding or endoscopic retrograde cholangiography (ERC). MRCP had a high accuracy in diagnosing acute cholecystitis with the finding of intramural high signal intensity. MRCP is an excellent method to evaluate acute biliary disease and may replace CT and ERC in the preoperative evaluation of acute cholecystitis. PMID- 9847626 TI - [MR cholangiography of congenital biliary malformations in infancy]. AB - We presented MR cholangiography (MRC) of congenital biliary malformations in infancy. MRC was obtained during induced sleeping. In two cases of congenital dilation of bile duct, MRC revealed cystic or spindle dilatation of intra- and extra hepatic bile ducts. In one biliary atresia, MRC revealed the serpentine gall bladder and cystic dilatation of the extrahepatic bile duct without connection to the dilated hilar bile duct. MR cholangiography, which can be obtained noninvasively, is useful for the diagnosis and the preoperative assessment of congenital biliary malformations in infancy. PMID- 9847627 TI - [Environmental endocrine disruptors]. AB - This review outlines the historical background of environmental endocrine disruptor issues and the particular sensitivity of the developing animals to exposure to estrogenic agents in the induction of longterm changes in reproductive and nonreproductive organs, and research needs of adverse effects of endocrine disruptors in experimental animals, wildlife and humans. Many chemicals released into the environment disrupt the endocrine system in wildlife and humans, and many of which have estrogenic activity by binding to the estrogen receptor. The animal and cell culture models can be utilized as an indicator of possible consequences of exposure to environmental endocrine disruptors. In humans, although the causes are not clear, hypospadia increased twice from 1970' and sperm count decreased and testicular cancer incidence increased. Epidemiological studies are needed to clarify the cause of these abnormalities in humans. More attention should be paid to abnormalities in genital organs exposed to endocrine disruptors during fetal and early postnatal development in wildlife, experimental animals and humans. PMID- 9847628 TI - [Trends in genome informatics]. AB - Genome Informatics is not only a new area of computer science for genome projects but also a new approach of life science. As the genome projects proceed, genome informatics is becoming more important to bio-industry as well as life science. The major subjects are as follows; Database technologies for integration of various kinds of biological data Knowledge discovery from the integrated databases Interpretation and analysis of DNA sequence data with the databases Computer technologies for simulating life system with knowledge extracted from the databases in order to check the validity of the knowledge In this article, the history and trend of development of computer technologies for the subjects are described except for computer simulation. PMID- 9847629 TI - [Novel function of Calcineurin--multipotential factor as protein a phosphatase]. AB - Calcineurin, serine/threonine phosphatase2B, is well known as a target of immunophilin-immunosuppressant complex such as cyclophilin-cyclosporinA and FKBP FK506. It has been disclosed that Calcineurin is involved in interleukin 2 gene activation pathway lead to T lymphocyte proliferation, however, its functions as a multipotential factor still remains unknown. Here we mention about a new aspect of Calcineurin-involved pathway through its direct interaction to Bcl-2, an apoptosis suppressor. This direct binding of Calcineurin to Bcl-2 results in blockage of KFAT4 nuclear import by the prevention of Calcineurin-targetted dephosphorylation of NFAT4. Moreover, the tight binding between Calcineurin and Bcl-2 facilitate Bcl-2 activation as a apoptosis inhibitor through dephosphorylation of phosphorylated form of Bcl-2 serving to apoptosis regulation. PMID- 9847630 TI - [Asthma and rhinoconjunctivitis caused by castor bean dust]. AB - A 57 years old trader of agricultural goods with chronic asthma had attended our medical department for diagnosis and therapy. Since more than 35 years the patient had always experienced work-related asthma episode and rhinoconjunctivities when he worked in workplace where castor-bean oil cake as fertilizer was sold. With the intensive contact with castor-bean containing goods beginning about eight years ago, his asthma attack had become chronic. The allergological diagnosis confirmed that he had high-degree type I-allergy to castor-bean allergens. PMID- 9847631 TI - [Almitrin and patients with COPD--a dangerous combination?]. PMID- 9847632 TI - [Long-term therapy of alpha 1-antitrypsin-deficiency-associated pulmonary emphysema with human alpha 1-antitrypsin]. AB - alpha 1-antitrypsin (alpha 1-AT) deficiency is a genetic disorder characterized by low serum levels of alpha 1-AT and a high risk of pulmonary emphysema at a young age. The resulting surplus of proteases, mainly of neutrophil elastase, can be balanced by i.v. augmentation with alpha 1-AT. However, it is not clear if affected patients benefit from long-term augmentation therapy and no long-term safety data are available. We examined 443 patients with severe alpha 1-AT deficiency and pulmonary emphysema receiving weekly i.v. infusions of 60 mg/kg body weight alpha 1-AT in addition to their regular medication. The progression of the disease was assessed by repeated lung function measurements, particularly the decline in forced expiratory volume in 1 second (delta FEV1). 443 patients with alpha 1-AT deficiency tolerated augmentation therapy well with few adverse reactions. The delta FEV1 in 287 patients with available follow-up data was 57.1 +/- 31.1 ml per year. Stratified for baseline FEV1, the decline was 35.6 +/- 21.3 ml in the 108 patients with an initial FEV1 < 30% and 64.0 +/- 26.4 ml in the 164 with 30% < FEV1 < or = 65% of predicted normal (p = 0.0008). The remaining 15 patients had an initial FEV1 > 65%. Long-term treatment with i.v. alpha 1 antitrypsin in patients with severe alpha 1-Pi deficiency is feasible and safe. The decline in forced expiratory volume in one second is related to the initial forced expiratory volume in one second as in alpha 1-antitrypsin deficient patients not receiving augmentation therapy. PMID- 9847633 TI - [Mendelson syndrome in infancy and childhood]. AB - Acute pulmonary failure caused by gastric acid aspiration is designated as Mendelson's syndrome. It is characterized by a trias of symptoms comprising bronchial obstruction, pulmonary oedema, and right ventricular failure. The pathomorphological pulmonary alterations show the typical symptoms of ARDS and allow the differentiation of three phases. The initial phase of injury is characterized by cauterization of the bronchial and alveolar epithelium. It is followed by the exsudative second phase during which alveolar oedema are developing. They impair the pulmonary surfactant synthesis and the formation of hyaline membranes. Fibrosis processes are typical of the proliferative third phase. Every of the mentioned three phases may be classified by their corresponding clinical symptoms. The therapy is entirely symptomatic and follows the intensive medical standards of the ARDS-therapy. PMID- 9847634 TI - [Magnetic resonance tomography with inhalation of polarized noble gases: new perspectives in functional imaging diagnosis of emphysema]. AB - PURPOSE: Based on a review of the background of MRI using inhaled hyperpolarized noble gases first experiences and perspectives for functional imaging in emphysema patients are presented. MATERIAL AND METHODS: In vonventional MRI, the spin density of protons, which is defined by the Boltzmann equilibrium, is the source of the imaging signal. Since proton density in the lungs is low and multiple air-tissue interfaces exist, MRI of the lung parenchyma is unsatisfactory. The possibility to artificially enrich the spin density (hyperpolarization) in noble gases (H3-3, Xe-129) by optical pumping results in a non-equilibrium polarization five orders of magnitude higher than the Boltzmann equilibrium. Hyperpolarized noble gases can then be applied as "inhaled contrast media" using a dedicated application device. At the MR unit several prerequisites must also be fulfilled: transmit-receive coil, boradband amplifier and fast sequences with low flip angles. These are essential for dynamic scans in breath hold tecnique of the highly diffusible He-3 or the well soluble Xe-129. RESULTS: He-3 and Xe-129 have been successfully applied for imaging of the ventilated airspaces. Besides the well-known narcotic effects of Xenon no adverse effects were observed. A homogeneous distribution of signal intensity can be regarded as a normal findings in people without lung disease. Obstructive diseases and emphysematous changes lead to generalized or localized signal inhomogeneities. Most likely they are caused by disorders of the distribution of ventilation bases on a different functional vehavior of different alveolar regions. By making use of the paramagenetic properties of oxygen, He-3 can also be used for local measurements of oxygen partial pressure in the lung. Xe-129 exhibits a different chemical shift within alveoli, interstitial space and vessels which can be measured by MRI. CONCLUSIONS: MRI using inhaled hyperpolarized noble gases is a functional imaging modality with high spatial and/or temporal resolution. First studies for early detection of obstructive lung diseases and disorders of distribution of ventilation in emphysema are promising. PMID- 9847635 TI - [Chemotherapy of advanced non-small-cell and small-cell bronchial carcinoma with bendamustine--a phase II study]. AB - In the palliative treatment of advanced SCLC and NSCLC there is a big need for effective and well tolerable drugs. Bendamustin is an alcylatic agent which had shown activity in the treatment of Non Hodgkin- and Hodgkin-Lymphoma as well as in the treatment of solid tumors like Mamma-Carcinoma and Colorectal-Carcinoma. We treated 21 patients with NSCLC (5 pat. Stad. III b, 16 pat. Stad. IV) and 22 pat. with Extensive Disease SCLC with Bendamustin 70 mg/m2 i.v., day 1-4 (q.28 days). We observed a response rate of 40.9% in the patients with SCLC (9 PR/40.9%), (0 CR) and no response in the patients with NSCLC. Hematologic toxicity in both groups was mild (Leucopenia WHO 1 + 2: 13 pat./30.2%, WHO 3: 2 pat./4.6%; Anemia WHO 1 + 2: 13 pat./30.2%, WHO 3: 1 pat./2.4%; Thrombopenia WHO 1 + 2: 4 pat./9.6%, WHO 3: 1/2.4%). Non Hematologic toxicity consisting of Nausea/Vomiting (WHO 2 + 3:13 pat./30.2%), Diarrhea (WHO 2 + 3:3 pat./7%), Obstipation (WHO 1 + 2: 2 pat./4.6%), Fever (WHO 1 + 2: 9 pat./20.9%) and Alopecia (WHO 1 + 2: 13 pat./31.7%, WHO 3: 1 pat./2.4%) was well tolerable. Cardiac Arrhythmias occurred in 7 pat./16.3% and PNP in 2 pat./4.6%. Treatment had to be stopped in one patient because of an allergic skin reaction. Bendamustin is a well tolerable cytostatic drug with a remarkable activity in advanced SCLC which is comparable to other well known agents in the treatment of this disease. Because of the good toxicity profile a combination with other compounds might be feasible. In advanced NSCLC Bendamustin showed no activity. PMID- 9847636 TI - [Perception of stress by mothers of children with language development disorders]. AB - Subject of this study is the maternal stress as a result of the disabilities of language impaired children. The report concentrates on the influence of the children disabilities and the maternal coping strategies on the stress in the families. The study is based on an inquiry of 98 mothers with speech and language impaired children. Nearly 50% of the mothers suffer from stress because of their children' disabilities. Special caregiving demands, family conflicts, anxiety about the child future, depressive mood, aggressive feeling or deception were often experienced by the mothers as a direct consequence of their child's problems. The correlation of this specific maternal stress with general stress factors was insignificant. The maternal stress was more due to the attention deficits and the behavioural problems of the children than to the language delay. An increased level of stress was hardly related to the severity of the impairment, but to the maternal coping strategies. Mothers applying emotion focused coping strategies experienced more stress than those who predominantly used problem focused strategies. The results prove that mothers of children with even minor disabilities feel overstrained by their children' disorders. During counseling experts should attach more importance to deal with the needs and wishes of mothers and families of language impaired children. Reduction of family stress can be achieved by intervention programs helping the mothers to improve their psychological resources. PMID- 9847637 TI - [Familial coping with cancer in childhood and adolescence. Possibilities for standardized assessment with a self-assessment method: results of an empirical pilot study]. AB - 44 parents of pediatric oncology patients answered mailed questionnaires about their own coping and about their sick children's coping and bodily complaints during oncological therapy. Most of the families coped well from parents' view. The dominant coping style of the parents consisted of active, problem-focused coping strategies in combination with an optimistic basic attitude, maintainance of family cohesion and belief in god or search for a meaning of the illness. The children and adolescents used predominantly adaptive coping strategies. The parental problem-group with a predominantly maladaptive coping-style reported low family cohesion, high depression, high wishful thinking and minimization. The maladaptive children and adolescents were rated as more socially withdrawing, more irritable, less optimistic and competent as well as less compliant. Strong correlations were found between parents' and their children's coping. Problem families with a special need for psychosocial support can early be identified by the coping questionnaires used in this study. Prospective longitudinal studies with multi-methodal and multi-perspective designs should be conducted to reach a better understanding of the family coping process in case of childhood and adolescence cancer. PMID- 9847638 TI - [Social intelligence deficits in autistic children and adolescents--subjective theories of psychosocial health care professionals]. AB - The paper is concerned with personal theories of health care professionals about deficiencies in social intelligence of autistic persons. In the component-model of social intelligence means the ability of individuals or groups, to interact with each other in social situations. This contains social perception, social behavior as well as social conceptions and refers to emotional, cognitive and normative aspects. 33 interviewees, working as psychologists or teachers in kindergartens, schools or therapy institutions, are questioned by a half standardized single interview concerning their beliefs about nonverbal social abilities, social perspective taking, and construction of a theory of mind in autistic persons. The major finding is: The impairments can be found in all aspects of social intelligence. Especially emotional handicaps, which are quoted by more than 80% of the interviewees, and low cognitive preconditions of mastering social stimuli, which are quoted by nearly all interviewees, are relevant. The subjective theories of the interviewees are in accordance to the models of parents as well as the models of the leading experts. The professional relationship to autistic persons and the practical experiences of the health care professionals lead to their specific personal theories of deficiencies in social intelligence of autistic people with wide consequences in respect to the professional contact with the autistic children and young adults. PMID- 9847639 TI - [Evaluation of child guidance counseling: status of research and references for future research]. AB - The evaluation of child guidance is placed in the context of quality control with emphasis on the significance of evaluation under field as opposed to laboratory conditions as well as the advantages of experimental or quasi-experimental field research over catamnestic studies. Up until now only three experimental evaluation studies have been carried out in the field of child guidance under natural conditions. These studies are first described in terms of their methods and results and then placed in a larger context on this basis. Proceedings from these studies based on methodological considerations and empirical evidence, comments and recommendations will be made concerning methodologically satisfactory and practically significant future evaluation research. They emphasize in particular the different evaluation strategies as well as various aspects of validity. PMID- 9847640 TI - [La Revista Espanola de Anestesiologia y Reanimacion. Motive for a study]. PMID- 9847641 TI - [Effectiveness of topical anesthesia enhanced by sedation and analgesia for cataract surgery]. AB - OBJECTIVES: To compare the efficacy of topical anesthesia and retrobulbar anesthesia for cataract surgery by lens emulsification. PATIENTS AND METHODS: Two hundred sixty patients were randomized to two groups in this open clinical trial. Patients with cataracts that could not be treated by lens emulsification were excluded. Group I patients (n = 129) received 0.5% tetracaine drops and intravenous fentanyl and propofol, along with continuous sedation. Group II patients (n = 131) received 2% lidocaine in the retrobulbar space and hypnotic doses of intravenous propofol before retrobulbar injection. The anesthesiologist evaluated anesthesia negatively if SpO2 was 90% and either heart rate or blood pressure varied more than 20%. The ophthalmologist evaluated anesthesia negatively if the eye did not remain fixed in the center, if blepharospasm appeared or if the anterior chamber of the eye collapsed. The patient reported the intensity of any discomfort experienced on a six-point scale. Anesthesia was determined to be effective when favorable evaluations were given by both the anesthesiologist and the ophthalmologist and when no significant discomfort (first three points on the scale) was reported by the patient. The two treatment groups were compared using a single and multiple factor analysis. RESULTS: Group II experienced significantly fewer instances of ineffective anesthesia than did group I (8 versus 22) and fewer negative evaluations by the ophthalmologists (7 versus 18). More patients in group I reported discomfort than in group II (46 versus 9), although most complaints were of slight discomfort. Multiple factor analysis showed that a patient in group I had 4.64 more chances of experiencing ineffective anesthesia. CONCLUSIONS: Topical anesthesia is less effective than retrobulbar anesthesia for cataract surgery by lens emulsification. PMID- 9847642 TI - [Characterization of dose profile of remifentanil with computer simulation: comparative study with fentanyl and alfentanyl]. AB - OBJECTIVES: To estimate the optimum dosing regimen and delivery system for remifentanil, a new opioid, using computer simulations based on information from pharmacokinetic and pharmacodynamic models available for fentanyl, alfentanil and remifentanil, as well as from clinical trials of fentanyl and alfentanil. PATIENTS AND METHODS: We estimated the site concentration ranges likely to be needed to blunt response to anesthetic or surgical stimuli and to recover from spontaneous ventilation. Dosing guidelines for remifentanil, fentanyl and alfentanil were estimated for three methods of administration (bolus, bolus + variable continuous infusion or constant continuous infusion). To that end, the time course of opioid concentration was simulated for hypothetical balanced anesthesia lasting 60 min. We then studied the number of boluses, the number of infusion rate steps, time taken to reach the terapeutic threshold, and time from turning off the infusion until reaching a concentration compatible with spontaneous ventilation. RESULTS: The estimated "effect site" concentration ranges for remifentanil were 6 to 10 ng.ml-1 during intubation; 4 to 6 ng.ml-1 during cutaneous incision; 4 to 7 ng.ml-1 for maintenance; and less than 2.5 ng.ml-1 for recovery of spontaneous ventilation. Simulated bolus administration indicated that 21 boluses of remifentanil, 4 boluses of fentanyl and 7 boluses of alfentanil were needed during one hour. The therapeutic threshold was reached within the first minute with remifentanil, within 2 minutes with fentanyl and within 1 min with alfentanil. Time until recovery of spontaneous ventilation was 7 min with remifentanil, 22 min with fentanyl and 14 min with alfentanil. In the simulation of bolus plus variable infusion, the initial bolus of remifentanil was 100 micrograms, the infusion rate for induction and maintenance was 25 micrograms.min-1 and the maintenance rate was 15 micrograms.min-1. The initial bolus of fentanyl was 300 micrograms, the infusion rate for induction and maintenance was 5 micrograms.min-1. The initial bolus of alfentanil was 2,000 micrograms, the infusion rate for induction was 200 micrograms.min-1 and the maintenance rates were 75 and 25 micrograms.min-1. The therapeutic threshold was reached in 1 min with remifentanil, in 2 min with fentanyl and within 1 min with alfentanil. Spontaneous ventilation was recovered 4 min after turning off the infusion of remifentanil, 4 min afterwards with fentanyl and 6 min afterwards with alfentanil. The simulated constant infusion rate for remifentanil of 15 micrograms.min1 (8 micrograms.min-1 for fentanyl and 75 micrograms.min-1 for alfentanil) allowed the therapeutic threshold to be reached in 10 min with remifentanil, in 22 min with fentanyl and in 17 min with alfentanil. Recovery of spontaneous ventilation occurred 5 min after closure of the infusion pump with remifentanil (24 min with fentanyl and 17 min with alfentanil). CONCLUSIONS: Information from pharmacokinetic and pharmacodynamic models allows us to establish the effect site concentration ranges for remifentanil and determine the ideal administration technique for this drug. The simulation also allows us to compare the properties of remifentanil to those of other common opioids such as fentanyl and alfentanil. The results are fairly consistent with clinical evidence, demonstrating the power of pharmacokinetic and pharmacodynamic models for rationally establishing opioid dosing guidelines. PMID- 9847643 TI - [Treatment of the hypertensive response during coronary surgery: comparison of high-dose propofol to propofol with nitroprusside]. AB - OBJECTIVE: To determine the efficacy of high doses of propofol for controlling hypertension during coronary surgery and to compared cardiovascular stability with propofol to that observed under lower doses of propofol with nitroprusside. PATIENTS AND METHODS: Forty patients were studied prospectively. The patients had good ventricular function and were scheduled for coronary surgery, randomized to two groups. Group P (n = 20) received 0.3 mg/kg propofol plus a 10 mg/kg/h perfusion. Hypertensive responses were treated with boluses of 0.3 mg/kg of propofol and progressive increases in the perfusion dose of 2.5 mg/kg/h at intervals of 2 minutes (maximum 15 mg/kg/h). If hypertension persisted it was treated with nitroprusside. Group N (n = 20) received propofol in perfusion at a dose of 8 mg/kg/h and hypertension was controlled directly with nitroprusside. During extracorporeal circulation, the propofol dose was reduced to 3 mg/kg/h in both groups and was adjusted in response to changes in arterial pressure, with nitroprusside added as needed. We recorded the number of patients becoming hypertensive during sternotomy and mediastinal dissection, the maximum doses of propofol and nitroprusside and the time taken to achieve control of hypertension. Arterial pressure and heart rate were recorded at intervals of one minute throughout the operation. RESULTS: Eight patients in group P and eleven in group N suffered hypertension (NS). Increasing the dose of propofol in group P controlled arterial hypertension in one patient. We found no significant differences between groups in amount of nitroprusside needed or time taken to bring episodes under control. Differences between the two groups in rates of intraoperative hypertension (65% in group P and 85% in group N) and hypotension (75% in group P and 55% in group N) and in duration of episodes were not statistically significant. CONCLUSIONS: Using high doses of propofol rather than moderate doses in combination with nitroprusside in coronary surgery does not improve control of either hypertension or hemodynamic stability. PMID- 9847644 TI - [Cardiocirculatory effects of intravenous anesthetic induction in an experimental model of acute hypovolemia]. AB - OBJECTIVE: To evaluate the cardiovascular effects of ketamine, midazolam, thiopentone and propofol in acutely hypovolemic pigs and to determine whether the association of ketamine and midazolam offers any advantage. PATIENTS AND METHODS: Twenty-two Landrace-Large-White pigs. After monitoring was begun, acute hypovolemia was induced by means of rapid exsanguination of 30% of calculated volume. Hemodynamic variables were measured: a) at baseline; b) after exsanguination; c) 2 min after anesthetic induction; d) 10 min after anesthetic induction, and e) after reinfusion of the exsanguinated volume. RESULTS: All pressures, cardiac output, cardiac index, and mixed venous oxygen saturation fell significantly with the induction of hypovolemia. Heart rate, systemic vascular resistances and arteriovenous oxygen differential increased. Ten min after anesthetic induction, heart rate in the midazolam group was significantly lower than in the ketamine-midazolam group. Arterial pressures decreased significantly after anesthetic induction with all drugs. The decrease in systolic arterial pressure was smaller in thiopenthal-anesthetized pigs than in pigs receiving either midazolam or propofol at the 10 min recording. The decrease in mean arterial and diastolic pressure after 10 min was smaller with thiopental than with any other drug. The decrease in mean arterial pressure was less in the thiopental and ketamine-midazolam groups than in the others after reinfusion. Diastolic arterial pressure at 10 min and after reinfusion had decreased less in the thiopental and ketamine-midazolam groups than in the propofol group. After anesthetic induction, the post-hypovolemic figures for cardiac output and cardiac index held steady or changes were slightly accentuated, with no statistically significant differences among the groups. CONCLUSIONS: The intravenous anesthetics evaluated were detrimental to cardiovascular function in acute hypovolemic pigs. Low-dose thiopental and ketamine plus midazolam may be the anesthetics of choice in this setting. Propofol caused the greatest degree of hemodynamic instability. PMID- 9847645 TI - [Original contributions of Latin Americans to anesthesiology]. AB - The original contributions of Latin American physicians to the science of anesthesiology are described. Many contributions have been unfairly ignored mainly because they were never published in English, but others have likewise been passed over even when published in the most prestigious journals in the field. Although many discoveries by Latin Americans have been made in the area of regional anesthesia, a considerable number of contributions have involved other aspects of anesthesia as well. PMID- 9847646 TI - [Postoperative complications in a case of iatrogenic hypothyroidism induced by diet drugs]. AB - Oral thyroid preparations, which were first administered in 1892 to treat myxedema, are the oldest effective endocrine medications. Since thyroxin became available, such drugs have been dispensed without a physician's prescription and have come to be mistakenly referred to as "diet pills". Self-prescription of such products can cause changes in thyroid function, with systemic repercussions. We report the case of a woman who took thyroid extracts, among other diet drugs to enhance weight loss without medical supervision. The patient failed to report taking such drugs during the preanesthetic visit. Interrupting therapy several days before surgery produced a state of hypothyroidism that led to marked postoperative hemodynamic changes necessitating transfer to the intensive care unit. PMID- 9847647 TI - [Anesthetic complications of familial Mediterranean fever]. PMID- 9847648 TI - [High-frequency jet ventilation in thoracic surgery]. PMID- 9847649 TI - [Notes on the methodology and basis for studying the psychological profiles of anesthesiologists]. PMID- 9847650 TI - [On Virginia apgar]. PMID- 9847651 TI - [Thyroid membrane puncture in a case of impossible tracheal intubation]. PMID- 9847652 TI - [The Cochrane collaboration]. PMID- 9847653 TI - [Prediction of mortality and quality of life in polytraumatized patients: APACHE II versus APACHE III]. AB - OBJECTIVE: To evaluate the APACHE II and III prognostic assessment systems as predictors of mortality in polytraumatized patients and to compare each system's admission assessments to the patient's quality of life one month, six months and one year later. PATIENTS AND METHODS: A prospective study of 130 polytraumatized patients admitted to the critical care unit was carried out. A polytraumatized patient was defined as "presenting two or more traumatic lesions that were immediately or eventually life-threatening". We studied age, sex, type of trauma, mortality, mean APACHE II and APACHE III scores during the first three days in the critical care unit for patients who survived and those who died, estimating relative risk of mortality by APACHE II and APACHE III, and quality of life one month, six months and one year after admission using Karnofsky scores (KPS). RESULTS: Mean patient age was 46.47 +/- 20.19 years; 78% were men and 22% women. Overall mortality was 20%. The most frequent trauma was craniocerebral (70.76%), followed by injuries ot the chest (53.8%) and extremities (46.15%). Mean APACHE II and APACHE III scores during the first three days were 10.22 +/- 5.33 and 32.75 +/- 16.42, respectively. Mean APACHE II and III scores were significantly higher (p = 0.02) in patients who died than in survivors (14.33 +/- 5.43 and 9.7 +/- 5.2, respectively, for APACHE II; and 43.27 +/- 17.68 and 30.16 +/- 15.79, respectively, for APACHE III). For each point increase of APACHE II or III scores, relative risk of mortality increased 1.09% (p = 0.03) and 1.02% (p = 0.03), respectively. Patients with APACHE II scores over 12 were 3.53 times more likely to die (p = 0.02). Patients with APACHE III scores over 35 were 3.05 times more likely to die (p = 0.02). One month after admission, 6.4% of patients had a KPS of 100 (normal, as before trauma), 35.8% achieved this score six months after admission and 82% after one year (p = 0.01). Mean APACHE III upon admission was significantly lower (p = 0.045) in patients who achieved KPS 100. CONCLUSIONS: The APACHE II and APACHE III prognostic systems predict relative risk of death in polytraumatized patients. APACHE III, but not APACHE II, at admission is significantly related to quality of life one year later. PMID- 9847654 TI - [Does the subdural space exist?]. AB - A potential space between the dura mater and the arachnoides is thought to exist, occupied by a serous fluid and called the subdural space. Recent studies may change this classical concept, however. The dura-arachnoid complex from the epidural to the arachnoid space is formed by morphologically distinct layers: the dura mater, the subdural compartment and the arachnoid mater, which are made up of different cell types. The dura mater consists of greater and lesser laminae formed mainly of collagen fibers aligned differently. The subdural compartment is formed by a number of so-called "neurothelial cells", which are in close contact with the inner dural layers. These cells are flat and have long interlaced branches. The arachnoides are made of cells grouped in three different layers. The outer layer is the "barrier arachnoid layer". Located just inside the anterior cell plane, this layer is made of less flattened cells that form an epithelial-type tissue, with complex cell-cell junctures surrounded by collagen fibers. The middle layer is the reticular arachnoid, composed of irregularly interlaced cells alternating with collagen fibers and intercellular gaps of varying sizes. The innermost layer, the trabecular arachnoid, is in direct contact with the subarachnoid space. The cells of this layer form strands that contribute to the weblike pattern found in the subarachnoid space. Recently, special techniques for fixing and preparing samples, preserving in situ the anatomical relations between the arachnoides and the dura mater, have allowed us to examine the normal configuration of the subdural space. All samples examined revealed the presence of a cellular plane between the dura mater and the arachnoides, with no evidence of the classically described space. The zone of least resistance in the dura-arachnoid complex was the subdural compartment, which could be torn mainly along intercellular spaces, though cell rupture was also observed, affecting the cytoplasmic membranes of adjacent cells. The subdural space is opened by tearing the subdural compartment between neurothelial cells alongside the collagen fibers of the dura mater. Such a tear can be caused mechanically by injecting air or contrast media, which exert pressure on a laminar structure that tends to separate because it is weaker than neighboring ones. PMID- 9847655 TI - [Continuous parascalene block for shoulder surgery]. AB - OBJECTIVE: To evaluate the efficacy and incidence of side effects of parascalene brachial plexus block for shoulder surgery and for postoperative analgesia. PATIENTS AND METHODS: This prospective descriptive study enrolled 186 patients scheduled for conventional (open) and arthroscopic shoulder operations. The parascalene approach to the brachial plexus was used, with an 18-G x 4.5 cm cannula (30 degrees bevel), applying the technique of detecting perforation of the prevertebral fascia. Mepivacaine 1.5% was administered for anesthesia, beginning with a test dose of 5 ml followed by doses of 25 ml. Bupivacaine 0.125% with 1/400,000 adrenaline was given for postoperative analgesia. Two groups were established according to type of surgery. In group A were patients undergoing open surgery and those with articular stiffness undergoing arthroscopic surgery; analgesics were delivered in continuous perfusion of 5 to 8 ml/h. Group B patients, who underwent arthroscopic surgery, received the analgesic in boluses of 10 to 14 ml every 4 to 6 hours. We assessed perception of perforation of the fascia, provocation of paresthesia, number of puncture attempts, anesthetic efficacy, postsurgical analgesia (visual analogue scale from 0 to 10) and complications. RESULTS: Blockade was effective for 155 of the 186 patients (83.33%). In 118 (63.44%) the perception of perforation of the prevertebral fascia was ostensible, in 52 (27.95%) it was slight and in 16 (8.6%) it was undetectable. Paresthesia was triggered by the cannula in 18 cases (9.13%) and by the test dose in 181 cases (97.31%). The mean number of puncture attempts was 1.2 +/- 0.09 per patient. During the early postoperative period analgesia was adequate for 92 (84.40%) of the 109 patients in group A. In group B, 45 patients (97.82%) required 1 to 2 analgesic doses (mean 1.4 +/- 2 doses) and one patient (2.17%) needed 6 doses within the first 24 hours. Catheters remained inserted as long as 36 hours to 10 days (mean 3.5 days) in group A. In group B they had been removed by 24 hours. Complications were transitory and slight. No cases of pneumothorax, no intra- or epidural anesthesia was required and no instances of vascular puncture occurred. CONCLUSIONS: The parascalene approach is easy to use and entails few risks while providing excellent anesthetic conditions for open and arthroscopic shoulder operations. It is useful for ambulatory analgesia to facilitate early rehabilitation after surgery. PMID- 9847656 TI - [Comparative study of preemptive and postincisional lumbar epidural morphine in pulmonary resection. Preliminary report]. AB - OBJECTIVES: To evaluate the efficacy and incidence of side effects of two types of lumbar epidural analgesia with morphine, preemptive or postincisional, combined with total intravenous anesthesia in chest surgery. PATIENTS AND METHODS: This double-blind prospective study enrolled 20 patients (ASA I-IV) undergoing lobectomy or pneumonectomy. Anesthetic induction and maintenance was provided with propofol, atracurium and alfentanil. Lumbar epidural analgesia (L2 L3) with morphine was provided for group A patients with 2 to 4 mg upon excision of tissue and for group B with 2 to 4 mg during anesthetic induction. The following variables were recorded: arterial blood gas concentrations, heart rate, SpO2, EtCO2, postanesthetic recovery, arterial gases, side effects and pain on a visual analogue scale. Top-up analgesia was provided by intravenous metamizole and/or epidural morphine. For statistical analysis we used ANOVA, chi-square tests and Student-Newman-Keuls tests. RESULTS: The need for propofol and alfentanil during anesthesia, and for morphine and metamizole after surgery were statistically greater in group A. Pain 18 hours after surgery was also greater in group A. No significant differences between groups for other variables was observed. CONCLUSIONS: Preemptive analgesia with lumbar epidural morphine in addition to the general anesthesia described here seems to provide higher-quality analgesia with few side effects, reducing the need for propofol and alfentanil during surgery and for postoperative morphine and metamizole. PMID- 9847658 TI - [Cardiorespiratory failure and diabetic autonomic neuropathy: a case report]. AB - Advanced autonomic neuropathy in diabetic patients is associated with homeostatic cardiovascular response. Sympathetic compensation diminishes and the organism becomes more susceptible to the depressant effect of certain drugs. The incidence of undesirable cardiovascular effects (bradycardia, hypotension and sudden heart failure) during anesthesia is higher in such patients. We report the case of a woman with diabetes who suffered two episodes of sudden heart failure after brachial plexus block through the axilla, followed one year later by sudden heart failure during general anesthesia for kidney transplantation. The possible contributing mechanisms are described. PMID- 9847657 TI - [A new hydroxyethyl starch for volume replacement: Elohes 6%]. AB - Various consensus groups convened in recent years to discuss plasma volume expansion solutions have suggested limiting the use of human albumin because of its high cost and have favored synthetic crystalloids or colloids for most clinical settings. Dextrans are colloids that are not widely used in most Europeans countries. Gelatins, in spite of the fact that unlimited amounts can be used, produce only moderate volume expansion and can trigger allergic reactions. The availability in Spain of hydroxyethylstarches (HES), a new type of colloid, may significantly change volume replacement strategy. HES are modified natural polymers. Three types, with different initial molecular weights, are used in Europe: high molecular weight HES, whose use in increasingly rare; low molecular weight HES (Expafusin), whose effect is short-lived; and medium molecular weight HES (Elohes, Fresenius-Laboratories Mein), which have recently been registered in Spain. Studies have shown that Elohes 6% (6% HES 200/0.62) provides volume expansion comparable to that of human albumin in clinical settings (cardiac surgery, shock, burns, etc.). The side effects of HES are usually minor. The frequency of anaphylactoid reactions is low, similar to that associated with human albumin. The effects on coagulation depend on molecular weight and duration of HES administration. Only in studies of hemodilution lasting 10 days with 6% HES 200/0.62 has VIII/von Willebrand complex been shown to decrease. If the recommended daily dose of this HES is respected, however, coagulation disorders are minimal. The effect of HES on kidney function is at present a subject of controversy. Thus, thanks to its prolonged effect on volume and few side effects, medium molecular weight HES colloids are the ones most often recommended for use in anesthesia and postoperative intensive care. PMID- 9847659 TI - [Preoperative information and informed consent from surgical patients]. PMID- 9847660 TI - [Starting up a low-cost postoperative acute pain ward]. PMID- 9847661 TI - [Anesthesia mailing lists on the Internet]. PMID- 9847662 TI - [Diagnosis of Wolff-Parkinson-White Syndrome during ketamine anesthesia]. PMID- 9847663 TI - [Magnetic resonance images of airway permeability with a laryngeal mask]. PMID- 9847664 TI - [Initial symptoms and mode of neurological progression in spinocerebellar ataxia type 6 (SCA6)]. AB - Spinocerebellar ataxia type 6 (SCA6) is genetically defined as a group of SCA characterized by late-onset pure cerebellar ataxia clinically and by a small CAG repeat expansion in the gene encoding the alpha 1A-voltage-dependent-Ca channel subunit (CACNL1A4) on chromosome 19p13.1 genetically. We analyzed the initial symptoms and the mode of progression in this disorder on 25 genetically verified patients. The initial symptoms were recurrent episodes of transient vertigo (72%) or unsteady gait (28%). Neurologically, they showed apparent gaze-evoked nystagmus (92%), transient positional nystagmus (83%), and periodic alternating nystagmus (4%), in addition to cerebellar ataxia. In addition to these episodic symptoms, all patients developed progressive cerebellar ataxia over years. These fluctuating symptoms at the initial stage of the illness were clearly different from those of other SCA, rather overlapping with those of episodic ataxia type 2 (EA2), an allelic disorder of SCA6. The clinical similarity indicates that there might be a common mechanism at least in part causing these two disorders. The mode of progression and their neurological features were also presented. PMID- 9847665 TI - [Valproic acid relieved marked rigidity in three patients with end-stage parkinsonism]. AB - We applied valproic acid (VPA) on the rigidity of three parkinsonian patients, two with Parkinson disease and one with striatonigral degeneration. They were all at Hoehn and Yahr's stage V and showed marked rigidity. In these patients, effect of L-DOPA had become limited or increasing the dosage of L-DOPA was difficult because of its side effects. Parkinsonian symptoms were assessed by using motor score of Unified Parkinson's Disease Rating Scale. The degree of rigidity in these three patients was markedly decreased with 300-600 mg/day of VPA. The blood level of VPA ranged from 24.8 to 66.5 micrograms/ml, which was relatively low compared with the effective blood level as an anti-epileptic agent. Parkinsonian symptoms other than rigidity, and the increased deep tendon reflexes which were present in the patient with striatonigral degeneration were not affected by VPA. Reduction of L-DOPA intensified rigidity again which had been under control. Trials of VPA on parkinsonism have been reported from two groups (Price PA, et al. 1978; Nutt J, et al. 1979), neither of which has observed any benefit of VPA. The difference of their results and ours seems to depend on the stage of patients; their patients had mild to moderate symptoms, whereas ours were in the end stage with marked rigidity. Since the effect of VPA upon parkinsonism is limited to rigidity, the end-stage patients whose care is difficult due to severe rigidity may obtain the best benefit of VPA. VPA is considered to take effect by activating gamma-aminobutyric acid (GABA) system. Because GABA is a common inhibitory neurotransmitter distributed in the wide areas of the central nervous system, it is difficult to locate the action site of VPA with regard to the amelioration of rigidity. The stretch reflex loop in the spinal cord does not seem to be the action site because no change was noted in deep tendon reflex. GABAergic striatal efferent neurons do not seem to be the sole action site either, because parkinsonian symptoms were not affected except for rigidity. The vestibular nucleus which receives strong GABAergic afferents from cerebellar Purkinje cells is an efficient tonus regulator. Since suppression of the function of the nucleus is known to reduce rigidity, it is at least a candidate for the action site of VPA. But there is no direct evidence for this matter. The exact action site of VPA remains to be elucidated. PMID- 9847666 TI - [A case of pure agraphia due to left parietal lobe infarction]. AB - We reported a case of a 63-year-old right handed man with pure agraphia due to the left parietal lobe infarction. The characteristics of agraphia in the patient were as follows. 1) The written letters were generally recognizable and well formed. 2) He succeeded in pointing to single Kana letter named by the examiner from the Japanese syllabary, but missed in pointing to Kana words. 3) Further, it took more time for the patient to point to even single Kana letter than for the control. 4) Most errors in Kana writing was substitution. Errors in Kanji writing are partial lacking or no response. But his ability in Kanji writing was facilitated by visual cues. He was unable to describe the Hen (a left-hand radical) and Tsukuri (the body) of some Kanji letters and to name some Kanji letters when their Hen and Tsukuri were orally given. We classified pure agraphia into two types out of some references. In one type (Type 1), letters in writing are poorly formed, but the ability to make words with the methods other than writing, for example spelling with anagrams or typing are preserved. In another type (Type 2), letters in writing were well-formed, but spelling with anagrams or typing were abnormal. Type 1 agraphia could result from the only deficit of graphic motor engram, while type 2 agraphia could be caused by the deficits other than graphic motor engram. Agraphia in this case belongs to the type 2. The features of agraphia in this case suggested that his agraphia was caused by a disorder in recalling graphemes of letters, and in arranging at least of Kana letters. PMID- 9847667 TI - [Long-term follow-up study with PET in a case of superficial siderosis]. AB - Long-term follow-up study with positron emission tomography (PET) has been conducted in a patient with superficial siderosis for ten years. A 63-year-old right-handed woman began to exhibit a cerebellar ataxia when she was 52 years old. Thereafter she has been exhibiting a slowly progressive course of pyramidal signs, hearing loss, anosmia, bilateral sciatica and memory disturbance in addition to the cerebellar ataxia. Series of x-ray CT and MRI disclosed a progressive atrophy of the cerebellum, specifically in the superior vermis, and a mild diffuse atrophy in the cerebral hemispheres, whereas no significant atrophy was seen in the brainstem. A marked hypointensity was seen along the rim of the brain structures including brainstem, cerebellum, sylvian fissures and the medial temporal lobes on MR T 2-weighted images. This hypointensity was also found at the edges of the third and fourth ventricles. These findings were regarded as haemosiderin deposit. By using oxygen-15, PET was carried out repeatedly with ten year interval. In the initial PET study, both cerebral blood flow (CBF) and cerebral metabolic rate of oxygen (CMRO2) were mildly decreased in the cerebellar hemispheres and the occipital cortices. The follow-up PET study revealed a progressive reduction of CMRO2 in the brainstem, cerebellar hemispheres and temporal cortices including the hippocampus in which the haemosiderin deposition was marked on MRI, whereas the reduction of CBF was not advanced as compared with CMRO2. It was suggested that the progressive reduction of CMRO2 in the brain sites where the haemosiderin deposition was observed on MRI reflected the clinical course of neurological deterioration. PMID- 9847668 TI - [Periodic alternating nystagmus in spinocerebellar ataxia type 6 (SCA 6)]. AB - We report a 61-year-old man with SCA 6 manifesting periodic alternating nystagmus (PAN). He first noted transient, recurrent episodes of "dizziness" at age 56. The symptom appeared several times a day, lasting for 3 years. At age 59, unsteady gait developed insidiously. On neurological examination, he showed limb and truncal ataxia, marked gaze nystagmus, and positional nystagmus. Thereafter, his ataxia became worse and PAN developed. Brain MRI showed pathologic atrophy which was confined to the cerebellum. PAN was rarely reported in spinocerebellar ataxia, being not described in SCA 6. The episodic "dizziness" at the initial stage has not gained much attention. Our observation indicates that the initial manifestation of SCA 6 overlaps with that of episodic ataxia type 2, an allelic disorder of SCA 6. PAN and fluctuating symptoms, as seen in our patient, must be carefully examined to see whether or not they can be parts of clinical presentations in SCA 6. PMID- 9847669 TI - [A case of late adult-onset dentatorubral-pallidoluysian atrophy (DRPLA) successfully treated with V-P shunt operation]. AB - A 44-year-old Japanese man was diagnosed as having late adult-onset dentatorubral pallidoluysian atrophy (DRPLA), whose CAG repeats in the DRPLA gene were 60 and 15. He developed gait disturbance, limb ataxia, pyramidal tract signs, dementia, and psychiatric symptoms including character changes within a few years of the above diagnosis. His T 2-weighted brain MRI showed symmetric high-signal lesions in the cerebral white matter and brain stem, in addition to cerebellar, brain stem, and cerebral cortical atrophy. Since the results of RI cisternography indicated that he manifested the clinical features of normal pressure hydrocephalus (NPH), V-P shunt operation was done. In a week after the operation, his gait disturbance, pyramidal tract signs, dementia and psychiatric symptoms were remarkably improved. White matter lesions have been thought to be concomitant with late adult-onset DRPLA patients, but some of these patients may have characteristics of NPH pathophysiology. PMID- 9847670 TI - [Ethics in clinical research]. PMID- 9847672 TI - [A review of the European community directives relating to materials and articles intended to come into contact with food products in the light of Polish legislation]. AB - The integration of Poland with the European Community makes the harmonization of Polish legislation necessary. Among numerous Directives, which provisions should be introduced into domestic legislation before Poland enters the European Union, the Directives related to materials and articles intended to contact with food consist a large group. The review and analysis of 17 Directives that appeared up to the end of year 1997 have been done. The list of these Directives is shown in the Table I. The framework Directive 89/109/EEC on the approximation of the laws of the Member States relating to materials and articles intended to come into contact with foodstuffs have been comprehensively discussed together with the specific Directives concerning the following matters: symbol which should accompany the materials and articles intendet to contact with food: Directive 80/590/EEC; plastics materials: Directive 90/128/EEC with later ammendments provided by the Directives: 90/128/EEC, 93/9/EEC, 95/3/EC and 96/11/EC; vinyl chloride monomer (acceptable limit and analytical method): Directives: 78/142/EEC, 80/766/EEC and 81/432/EEC, regenerated cellulose film-Directives: 93/10/ECC i 93/111/EEC; ceramic articles: Directive 84/500/EEC; testing migration of constituents of plastic materials-Directives: 85/572/EEC and 82/711/EEC with later ammendments: Directives: 93/8/EEC, 97/48/EC, The relevant Polish regulatory provisions concerning the matters related to materials and articles intendet to contact with foodstuffs have also been discussed. The need for harmonization of Polish regulations with the relevant legislation adopted by the Member States of the European Community was emphasized. PMID- 9847673 TI - Use of recycled fibers for paper and board in food contact. PMID- 9847674 TI - [A comparison of selected parameters of potatoes health quality from ecologically oriented and conventional farms]. AB - The aim of work was to compare nutritional quality and wholesomeness of the potatoes from ecological (ECO) and conventional (CONV) farms. Other aim was to check if the west-european data telling about better nutritional quality of ECO potatoes can be confirmed in Polish agricultural conditions. Nine ECO farms located in Torun and Sieradz provinces, having EKOLAND's certification were selected to study. A neighbour CONV farm with similar production conditions was selected to each ECO farm. ECO farmers manured potatoes with composted cow manure or compost giving on average 30 t/ha. CONV farmers used mineral fertilizers NPK in 1:1:1,5 ratio giving 100 kg N/ha. In the autumn several compounds were determined in potatoes: the contents of dry matter according to PN-88/R-04013, the contents of nitrates and nitrites according to Draft International Standard ISO, the contents of Pb and Cd by means of ASA method and the contents of vitamin C as L-ascorbic acid according to PN-90/A-75101/11. The ECO potatoes contained in both years significantly less nitrates than CONV potatoes. In 1992 the nitrates contents was much higher in both kinds of potatoes than in 1993. The nitrites contents was in both years higher in CONV potatoes but the difference was statistically significant only in 1993. The lead contents was low in both kinds of potatoes and didn't exceed permissible level. The cadmium contents was in 1992 significantly higher in CONV tubers than in ECO tubers and exceeded permissible level. In 1991 and 1993 both kinds of tubers contained similar cadmium levels. The contents of dry matter and vitamin C was similar in both kinds of tubers. The study has shown that ECO potatoes were more wholesome than CONV potatoes. The results obtained in Polish agricultural conditions confirmed west-european data. PMID- 9847675 TI - [A determination of iodides in salts: a validation of methods]. AB - The studies reported were aimed at finding a simple analytical method enabling quantitative determination of iodide in table iodised salt and in therapeutic iodide-bromide salts. The analytical procedure proposed is a modification of spectrophotometric method recommended in the Polish Standards. The method based on the reaction of iodide oxidation by sodium nitrite was validated by determining its precision, accuracy and linearity. Statistical analysis has shown that the coefficient of variation varies between 2.73 and 4.82%, recovery is from 99.17 to 101.83% and falls within the confidence interval for the mean recovery at the assumed level of significance. The method can be used for controlling the technology of table salt iodisation. PMID- 9847676 TI - [SO2 and NO2 air pollution measurement by passive monitors]. AB - Air pollution concentration measurement in Poland are routinely carried on using aspiration methods. This technique required suitable measurement point preparation and its monitoring. During last years passive monitors are used more and more frequently for some air pollutants concentrations measurement. The above mentioned method is recommended when spatial differentiation of air pollution are analized. Results of such measurements carried of in the area of Poznan are presented in this contribution. PMID- 9847677 TI - [The radon therapy: radon inhalation spa in Kowary]. AB - In this paper the measurements of radon-222 concentration in the air of Kowary Radon Inhalatorium in the vicinity of Jelenia Gora, Lower Silesia, Poland, performed by Radiation Protection and Radiobiology Department of National Institute of Hygiene in Warsaw, have been presented. For the measurements the integration method with the application of charcoal canisters have been used. The radon concentration was found within the very broad ranges from about 4 to 420 kBq m-3 depending of the season of the year. Basing on the radon concentration measurements during the inhalation therapy session and taking into account the equilibrium factor of 0.8 the doses to the bronchi of the patients in range from 1.2 to about 204 mSv (0.12-20.4 rem) have been assessed. The majority of patients (ca. 76%) received the doses between 115 and 204 mSv. PMID- 9847678 TI - [Exposure to radon-222 of inhabitants of buildings in Olsztyn]. AB - The concentrations of radon-222 in the air of some buildings in Olsztyn were measured. The main source of radon in buildings is in general the ground under building and the materials used for building structure. In this work the results of radon-222 concentration measurements in the air of some buildings constructed before the 1939 year, in the buildings constructed after 1945 year with the traditional use of the bricks and in the buildings constructed with the use of great prefabricated plates are presented. The relations between radon-222 concentrations in the basements and in the first floor flats situated above the basement were evaluated. Based on the mean radon concentrations in the air of the various types of buildings investigated the effective doses for the inhabitants of each type of buildings were estimated. PMID- 9847680 TI - [Some pharmacodynamic properties of the "Marysiennka" healing water from the Cieplice-Zdroj springs]. AB - On the ground of the performed experimental investigations it has been shown that the "Marysienka" water, given to rats intragastrically in a single daily dose of 14.3 cm3/kg body weight for the spans of 20 and 40 days and in the some spans of time as drinking water ad libitum, causes a statistically significant decrease of the levels of total cholesterol, total lipids, triglycerides, and an increase of the HDL fraction of cholesterol. Within the range of the investigated parameters of electrolytic economy a statistically significant fall of the level of calcium and decrease tendencies of sodium, magnesium and increase tendencies of potassium in the blood serum of the examined animals could be observed. After 20 days of the administration of the investigated water both intragastrically in a single daily dose of 14.3 cm3/kg body weight and in the animals that were receiving that water as drinking water ad libitum a marked compensated metabolic acidosis was observed. A long-lasting exposition of the rats (40 days) to that water may lead to the rise of compensated metabolic alkalosis. No essential influence of the studied water on the indicators of protein and carbohydrate metabolism and on the morphological composition of the peripheral blood and its smear was noted. The studied water shows no cholagogic effect in guinea-pigs, nor any diuretic effect in rats. PMID- 9847679 TI - [The measurement of radon in kindergarten and infant nursery buildings in East North Poland]. AB - The measurements of radon concentration in 58 buildings of day nursery schools and kindergartens were performed in the six province of the East North Poland. The measurements of indoor radon were performed by means of a charcoal canister (Pico-Rad). The concentration ranged from 4.5 to 320 Bq/m3 and have arthimetic average 24.5 Bq/m3, geometric average 15.7 Bq/m3, median 15 Bq/m3. The highest average values were observed in Suwalki Province and Olsztyn Province. The results of all these measurements do not exceeded the limit of the Polish law. PMID- 9847681 TI - [Dietary assessment of adults in three villages in Warsaw, Radom and Biala Podlaska districts]. AB - The objectives of this study was the assessment of dietary patterns of adults individuals living in three villages: Celestynow (Warsaw district), Jasionka (Biala podlaska district) and Babin (Radom district) in Poland. The study was conducted in winter 1994/95 among 71 men and 77 women aged 18 years or more. The information on food consumption during one weekday was obtained by 24-hour dietary recall method. On the average the percentage of total energy intake from proteins and fats was above the recommended limits whereas the proportion of energy derived from carbohydrates was slightly deficient. For women the average daily intake of most nutrients under study (with exception of animal proteins, fats, sodium, phosphorus, manganese, vitamin E, B2 and niacin) was below recommended levels. The consumption of carbohydrates (also dietary fiber), magnesium, zinc, copper, calcium was 29-35% below recommendations and case of vitamin C-51%. In general men had higher mean intake of energy and nutrients than women. Their average daily diet did not meet recommendation only for carbohydrates, dietary fiber, calcium magnesium, copper, vitamin C and A. Men consumed too much animal proteins, sodium, phosphorus and cholesterol. Daily consumption of minerals and vitamins from diet and supplements together for those subjects who used supplements (41% of respondents) met or exceed recommended dietary intake, with exception of potassium for women. Taking consideration number of faults founded in the nutrition patterns of adults from rural area more nutritional study in this subpopulation group should be conducted. PMID- 9847682 TI - [The effect of smoking on environmental exposure to carbon of inhabitants in highly and little industrially developed areas]. AB - The aim of the work was an estimation of the effect of smoking on environmental exposure to CO people living in industrial area of Sosnowiec (a city situated in Upper Silesia) and Tomaszow Lubelski (not much industrially town situated at the border of Roztocze National Park). The investigated material was vein blood of 102 blood donors from Sosnowiec and 103 blood donors from Tomaszow Lubelski both smokers and non-smokers. The level of CO exposure was estimated by determination of carbon monoxide haemoglobin (HbCO) concentration in blood by Wolf's method in Antczak's adaptation. It was stated that average HbCO concentration at non smokers from Sosnowiec (2.09%) was about 0.9% higher than the one at non-smokers from Tomaszow Lubelski (1.17%). An increase of HbCO concentration in blood of non smokers from Sosnowiec as compared with non-smokers from Tomaszow Lubelski included also 50 and 90 percentile. In relation to 50 percentile it was 2.05 and 1.0% and in relation to 90 percentile 2.4 and 3.7% respectively. The influence of external environment on CO exposure at smokers was estimated on the base of HbCO determination in blood before starting smoking. It was 2.77 at smokers from Sosnowiec and 1.41% at smokers from Tomaszow Lubelski. Analysing the effect of smoking on smokers exposure to CO it was stated that HbCO concentration in blood in both groups increased linearly with the increase of the number of smoked cigarettes, and average increase per one cigarette was 0.43% for smokers from Sosnowiec and 0.57% for smokers from Tomaszow Lubelski. On the base of the obtained results the following conclusions were drawn: 1. industrial and communal environment contamination with carbon monoxide influences considerably the exposure of both smokers and non-smokers. 2. smoking effects significantly more the exposure of people (smokers) to carbon monoxide than environmental pollution in the investigated industrial area. 3. the exposure to carbon monoxide at smokers up with the environmental exposure. PMID- 9847683 TI - [Low zinc levels in metabolic X syndrome (mzX) patients measured by hair zinc composition analysis]. AB - The hair zinc content in 16 patients with metabolic X syndrome (mzX) was measured by means of atomic absorption spectrometry method. The mean concentration (125.13 mg/kg) was lower than in the majority of other published studies. The differences among groups of patients with different sex or diseases (e.g. coronary heart disease, hypertension, type II diabetes mellitus) were not significant. It seems to be probably that deficiency of zinc plays a role in pathogenesis of mzX or that it is a consequence of mzX. PMID- 9847684 TI - [What is your diagnosis? Spontaneous rupture of the esophagus (Boerhaave syndrome) in chronic alcohol abuse]. PMID- 9847685 TI - [Cirrhosis of the liver as a precancerous condition]. AB - Cirrhosis of the liver can be regarded as premalignant state, since more than 80 percent of hepatocellular carcinoma (HCC) in the western world develop in a cirrhotic liver. The risk to develop this malignancy depends on the activity of the underlying cirrhosis, its etiology and the duration of the disease. Patients suffering from cirrhosis of the liver due to HBV-, HCV- or HDV-infection and patients with genetic hemochromatosis exhibit a high risk for HCC. This risk is further increased by cocarcinogens, such as alcohol, nicotine and toxins. Ultrasound and AFP-studies aim to diagnose HCC early. The sensitivity of AFP in the serum is remarkably low (about 64%). In contrast a normal AFP-concentration (< 20 ng/ml) carries a high negative prognostic value (> 90%). Patients suspected to suffer from HCC according to the results of screening procedures should be subjected to additional radiologic investigations, such as CT-arterioportography or lipiodol-angiography. PMID- 9847686 TI - [Hepatocellular carcinoma: npn-surgical treatment]. AB - The therapeutic modalities in patients with hepatocellular carcinoma (HCC) depend on the number, size and location of the lesions as well as the stage of the underlying liver disease and the physical condition of the patients. In patients with small and solitary lesions resection, liver resection and in selected cases liver transplantation can be curative. In more advanced stages of the disease with larger or multiple lesions PEI and/or transarterial chemotherapy with or without embolisation (TACE or TAC) can slow the progression of the disease. In disseminated disease, a radio-therapeutic approach can be taken in selected cases. The therapeutic strategy in patients with HCCs has to be individualized, frequently using a combination of therapeutic modalities. Different from the earlier unfavourable prognosis, today for most HCC patients a therapeutic strategy can be offered that results in a prolongation of life and in some cases even cure. PMID- 9847687 TI - [Hepatocellular carcinoma: surgical treatment]. AB - Worldwide the hepatocellular carcinoma (HCC) is one of the most common malignancies. There is a coincidence with liver cirrhosis or chronic hepatitis B/C in most cases. HCC can be suspected by ultrasound and by rise of the tumor marker (AFP). Further investigations, like biopsies, are not necessary if angiography or computed tomography in combination with patient history and elevated AFP levels are positive. The prognosis of untreated HCC is extremely poor. Live expectancy of symptomatic patients is only a couple of weeks. Radical tumor removal by liver resection or transplantation is the only treatment with curative intent. However these options are only suitable for patients with limited disease. Five year survival after curative liver resection depends on the tumor stage, ranging from 25% to 67%. The results after liver transplantation are similar for small cancer. Large, symptomatic tumors are in most cases only suitable for palliative treatment (chemoembolisation, ethanol injection, chemotherapy, immunotherapy). The strong dependence of prognosis on tumor extent underlines the importance of screening patients with elevated risk of developing an HCC. The early recognition of small tumors allows curative therapy with good results. PMID- 9847688 TI - [Hepatocellular carcinoma: interdisciplinary treatment concept]. AB - In most cases hepatocellular carcinoma develops in a cirrhotic liver. The stage of the cirrhosis and the number and size of the liver tumors are decisive for the prognosis and the individual treatment possibilities. For each therapeutic procedure there are specific contraindications which have to be considered. In a prospective comparative study of different established methods of treatment, i.e. hepatic resection, percutaneous ethanol instillation, transcatheter arterial embolisation and tamoxifen therapy, tumor response, survival time, complications and factors, determining prognosis are analysed. The therapeutic concept of the study can thereby be used as a model for the interdisciplinary treatment of hepatocellular carcinoma. PMID- 9847689 TI - [Acute hepatitis A with accompanying cholestasis]. PMID- 9847690 TI - [Collagenous colitis. Patient 1933, Swiss manager, presenting symptoms: watery diarrhea]. PMID- 9847691 TI - [Structure and dynamics of the transcription factor, Myb, in DNA-sequence recognition]. PMID- 9847692 TI - [Evolutionary molecular engineering of proteins]. PMID- 9847693 TI - [Bruton's tyrosine kinase: what's known now]. PMID- 9847694 TI - [Role of chemokine receptors in T lymphocytes]. PMID- 9847695 TI - [Signalling roles for selectins]. PMID- 9847696 TI - [Vascular development and gene regulation in endothelial cells]. PMID- 9847697 TI - [Mechanism of stress response and gene expression of glyoxalase I in yeast]. PMID- 9847698 TI - [Roles of C2 domains of synaptotagmin in the regulation of exocytosis]. PMID- 9847699 TI - Hypothermia in trauma patients. PMID- 9847700 TI - Hypothermia in trauma patients. AB - Accidental hypothermia (AH) can be defined as an unintentional decrease in core temperature below 35 degrees C during cold exposure by individuals without intrinsic thermoregulatory dysfunction. Pathophysiological changes can be attributed both to the severity of hypothermia and to co-morbid factors such as trauma, submersion, intoxication and underlying diseases. In trauma victims stratified according to the Injury Severity Score, the factor hypothermia is considered to be a poor prognostic sign for survival. In these patients rewarming therapy should be applied as soon as possible. In the Utrecht University Hospital, adult patients with AH are managed according to an algorithm based on their presenting hemodynamic conditions. Patients with perfusing cardiac rhythms and systolic pressures over 80 mmHg will receive continuous arteriovenous rewarming (CAVR). Arrested and hemodynamically instable patients are treated with cardiopulmonary bypass (CPB). In a 3-year period, 22 patients with AH were admitted to the emergency department. Fourteen patients had a trauma as the cause of hypothermia. Twenty patients were treated according to the algorithm (CAVR n = 18, CPB n = 2) and two patients were alternatively managed. Mortality in the CAVR group was 28% and total mortality reached 32%. Complications of CAVR are related to placement and removal of vascular catheters and may be severe in these patients with impaired coagulation. CAVR permits a good access to the (trauma-) patient and rewarming can be continued synchronously with diagnosis and treatment of various injuries. PMID- 9847701 TI - [Accident-induced pseudarthroses of the dens axis. Etiology, follow-up and therapy]. AB - In a retrospective analysis the significance of internal fixation of unstable and symptomatic non-union of the odontoid was evaluated. In all but one cases a type II fracture of the odontoid was the underlying cause for the pseudarthrosis. The time interval between trauma and definitive diagnosis varied between 6 months and 30 years. All patients asked for medical advice because of acute neck pain; in three cases additional neurological deficits were notified. In all but one case the pseudarthrosis was classified as unstable on flexion-/extension views. The occurrence of neurological deficits showed a positive correlation with the amount of dislocation of the pseudarthrotic odontoid. Nine out of 10 patients underwent internal stabilisation of the non-union. Reoperation because of persistent instability/failure of stabilisation had to be performed in two cases (22.2%). At follow-up all 9 patients were pain-free. The neurological deficits in two patients had improved significantly. According to our experience internal stabilisation in unstable non-union of the odontoid is recommended to achieve significant reduction of the pain level. Also improvement of the neurological deficits can be expected depending on the duration of the symptoms. PMID- 9847702 TI - [Results of posterior inferior capsular shift in posterior shoulder instability]. AB - PURPOSE: The purpose of this study was the evaluation of our results after posteroinferior capsular shift in patients with a posterior shoulder instability. MATERIALS AND METHODS: In a prospective study we evaluated 12 patients (14 shoulder joints) with recurrent posterior shoulder dislocations who were stabilized by a posteroinferior capsular shift procedure. For the clinical documentation the Rowe-Score, the ASES-Score and the SF-36 were used. Furthermore the patients were submitted to standardized EMG-, isokinetic and proprioceptive tests. RESULTS: After an average follow-up of 22.2 months there were two recurrences which occurred during the first postoperative year. The Rowe-Score improved from 46.25 to 76.34 points. The ASES-Score showed an average of 42.43 points pre-operatively and at the time of follow-up an average of 78.86 points. Most parameters of the ASES-Score as well as the score in total had a constant positive tendency. Only the subjective rating of stability showed a slight decrease after the first six months due to the two redislocations. But even the stable shoulder joints demonstrated limitations when throwing a ball. The SF-36 showed a definite gain in the overall quality of living as well. The EMG-tests at follow-up showed a deficit for the patients compared with a control group; even more for the female than for the male patients. Similar tendencies were found with the isokinetic strength. The proprioception did not show any differences between the patients and the control group. CONCLUSION: With adequate patient selection and after failure of all possible conservative treatments the posteroinferior capsular shift is a therapeutical option in recurrent posterior shoulder instability. PMID- 9847703 TI - [Distal, metaphyseal compression fractures of the radius. Results of open reposition, stable defect replacement with cortico-cancellous iliac crest bone and plate osteosynthesis]. AB - Conservative treated distal radius fractures with extended metaphyseal void and missing cortical support resulted in loss of reduction. From 1988 till 1994 117 fractures of the distal radius were treated by open reduction, filling the metaphyseal void with a corticocancellous iliac bone graft and plate fixation. In this retrospective study results after surgical reconstruction were evaluated radiologically and clinically and were discussed to conservative treatment. 30 patients suffering from extraarticular metaphyseal distal radius fracture were evaluated after a mean of 42 months (minimum 20, maximum 68 months) after surgery. Ulnar variance was restored in 70% to the uninvolved side. Ulnar minus variance up to 2 mm persisted in 30%. Radial joint angles were restored to normal in 53%. In 40% of the patients a loss of reduction up to 10 degrees and in 7% up to 25 degrees occured. Active range of motion in the sagittal and frontal plane was equivalent to the non involved side in 30%. 70% of the patients showed reduced range of motion up to 20%. Rotation was unlimited in 77%, reduced up to 10% in 23%. Subjective results were excellent in 90%, good in 10%. Compared to conservative therapie, surgery showed superior results. PMID- 9847705 TI - [Morbidity after contralateral transplantation of the patellar ligament for cruciate ligament replacement]. AB - After cruciate ligament reconstruction using an autogenous central bone-patellar tendon-bone graft persistent complaints are described in the literature. There is the difficulty to distinguish the morbidity associated with the graft harvest and the morbidity caused by the injury and the reconstruction procedure. In order to clarify the morbidity resulting from the graft harvest alone, we evaluated those patients who had transplantat harvest from the opposite knee. Between 1990 and 1995 a central bone-tendon-bone graft has been taken from the contralateral uninjured knee in 37 patients, in 22 cases for anterior and in 15 cases for posterior cruciate ligament reconstruction. 27 patients were clinically (IKDC Score), isokinetically and radiologically followed up in an average of 25.3 months after the operation (range: 9-73 months). 21 patients were classified into IKDC group A. Except of one patient with an extension deficit of 5 degrees all others showed normal range of motion. Only 5 of 27 patients expressed minimal knee pain during vigorous activities. Three of them reported about weather sensitivity in addition. These 6 patients were ranked in group B. No patient was classified into IKDC group C or D. The isokinetic examination showed a normal level of quadriceps strength. At final followup, the technique of Blackburne and Peel was used to assess patellar height. There was no radiographic evidence of patella contracture or baja. According to our results taking the autogenous CBTB graft used for cruciate ligament reconstruction leads to no serious morbidity. PMID- 9847704 TI - [How many severely injured multiple-trauma patients can benefit from the biomechanical advantage of early mobilization following femoral intramedullary nailing?]. AB - The benefit of early operative stabilization of femoral fractures is established in patients with multiple injuries. In the last few years the unreamed femoral nail is favoured for internal fixation of femoral fractures despite pathophysiological concerns. The foremost advantage of femoral nails compared with plate fixation is the possibility of early full weight bearing. The aim of this retrospective study was to investigate, under consideration of the severity of injury, the extent of injury, and the clinical course, if multiple injured patients with concomitant femoral fractures benefit from the preferred intramedullary nailing with early weight bearing. Three hundred and two (23.8%) out of 1271 multiple injured patients (ISS > 17) had a concomitant femoral fracture. Fourty-seven out of 302 patients were children under 16 years of age, remaining 255 patients. Eighteen out of 255 patients died within the first 21 days after trauma and 66 patients required mechanical ventilation for more than three weeks (171/255). Thirty patients suffered from severe head injury (AIS-head > 3) and seven from severe pulmonary contusion with concomitant abdominal injury (134/255). Two patients had grade III open femoral fractures with vascular injury. Ipsilateral unstable pelvic fractures were seen in 11 patients, seven patients had ipsilateral intraarticular femoral fractures, and ipsilateral intraarticular fractures of the lower leg or foot were observed in 40 patients (74/255). The results demonstrate, that only 74 (29%) out of 255 multiple injured patients (> 16 years of age) had a theoretical benefit of early weight bearing. Seventy percent of the patients did not benefit from intramedullary nailing considering full weight bearing. With regard to pathophysiological concerns alternative methods of fracture fixation should be discussed for these patients. Primary fracture fixation with external fixators and secondary internal fixation proved to be a save alternative method. The complication rate of plating is comparable to intramedullary nailing but associated with less severe systemic risks. Primary plating of femoral fractures would not delay mobilization of most multiple injured patients. PMID- 9847706 TI - [Injuries of the thoracic and lumbar spine]. PMID- 9847707 TI - [Current status and developments in vehicular safety of automobiles]. PMID- 9847708 TI - [Status of knowledge in expert assessment of occupational disease 2108-2110]. AB - Necessary precondition for acceptance of spine disorders as occupational disease is an excessive occupational exposition, its compatibility with and and an explicit causal relation to this disorders. The latter may be assumed in case of multisegmental spondylosis of the column, combined with local impairment of function and pain, if the vertebral disorders of the occupationally exposed column-segment are significantly worse than those of other vertebral segments. Finally non-occupational, preexisting discrete vertebral disorders (e.g. scoliosis, spondylolisthesis) have to been excluded. So far there are no alternatives or rejections to this consensus, called "Hamburg formula". It should be used as long as no other valid, positive markers of occupationally caused disorders can be defined on the basis of reliable epidemiological investigations. PMID- 9847709 TI - Is psychiatric nursing research barking up the wrong tree? PMID- 9847710 TI - Scaffolding student learning in clinical practice. AB - For many years the profession has acknowledged a schism between theories taught in the classroom and the practice of clinicians. This tends to arise from beliefs that knowledge which has been generalized (formalized) can be readily transferred to informal settings (practice). Whilst apprehension of formalized knowledge is crucial to professional development, a mediator is necessary to demonstrate its relevance to practice. A variety of solutions have been offered which all have value but without effective social support fail to resolve the problem. Findings from a longitudinal study investigating the professional development of pre registration nursing students indicate the importance of sponsorship by a member of clinical staff and participation in legitimate peripheral activities. In the absence of effective sponsorship, students found it difficult to participate in clinical activities or to learn. As a result, their professional development during placements became stunted with subsequent implications for their professional future. Questions concerned with this phenomena were explored using documentary evidence supplied by research participants, observations of their practice and focused interviews throughout their programme. Sociocultural or activity theories of human learning offer some attractive explanations. One strategy is scaffolding which takes place within sponsored nursing activities and builds on the important concept of the Zone of Proximal Development (ZPD), where speech becomes a tool to mediate learning and development. As a diagnostic tool, scaffolding enables both supervisor and learner to recognize knowledge-in-waiting and knowledge-in-use and hence learning need, thus stimulating opportunities for further learning and professional development beneficial to both learner and supervisor. It provides a means for theory and practice to become integrated. PMID- 9847711 TI - Nursing research--taking an active interest. AB - This paper discusses the issues raised by the still unfolding transformation of nurse education attitudes to research, from the traditional stance--that it was something that other people do--to the realization that quality research is central to the development of the profession and its competitive survival into the 21st century. In particular, the problems and challenges presented by the Higher Education Funding Council for England research assessment exercise, and the policy of the funding council, are examined. Research in schools of nursing is reviewed under the headings of funding, contracts, support, discipline, publication and staff recruitment and retention, to attempt to identify those approaches most likely to yield a research output of an acceptable volume and quality. PMID- 9847712 TI - Defining and assessing competence: the distraction of outcomes and the importance of educational process. AB - The concept of competence is in common usage in various areas of education and training at present, and it is evident from these different usages that it has a variety of meanings. These meanings are explored, with particular attention being paid to the increasing influence of the National Vocational Qualification (NVQ) initiative and the rise of vocational education. It is concluded that the utilization of competence within these fields offers little to nurse/midwifery education (with the exception of a refocusing on the concept of skills) because it is exactly that: education, and is therefore different in some significant aspects from training and vocational approaches. It is suggested that a holistic conceptualization of competence is the most appropriate for nurse/midwifery education: one that empowers students and practitioners, facilitating critical thinking, rather than emphasizing, as the NVQ initiative does, outcomes over the importance of educational process and the use of a variety of forms of knowledge in practice. It is acknowledged that there is probably a skills deficit in some areas of nursing/midwifery education, although definition of this term 'skills' remains problematic, and that this should be addressed through the increased use of skills laboratories. PMID- 9847713 TI - Facilitating the process of critical thinking for nursing. AB - An exploratory study was conducted during 1995 to examine the degree to which critical thinking was encouraged in nursing education throughout New South Wales (NSW), Australia. The study identified whether a sample of graduate nurses and nurse educators at 12 faculties of nursing in NSW shared similar ideas about what critical thinking entails, the best ways in which to develop critical thinking processes and whether critical thinking is a reasonable way for nurses to achieve skilled and effective nursing interventions. The findings indicate that both nursing students and nurse educators favour the facilitation of critical thinking for nursing for very practical reasons. These refer to improving professional standards of practice, stimulating inquiry and promoting sound reasoning in practice, as well as contributing to personal and professional development. Study participants were found to favour a variety of teaching and learning strategies for critical thinking, and this finding is the focus for this discussion paper. The majority of participants stated that nurses would perhaps be better able to abstract principles of thinking from the specific contexts in which they are practised. Strategies found effective for nursing practice included a variety of approaches: direct learning of skills that contribute to critical thinking, such as analysis; infusion, or integration of critical thinking in all areas of learning; and learning to think critically within distinct disciplines of thought. Analysis of the findings, therefore, suggests that critical thinking is thought to be an important component of nursing practice and that in nursing it is a complex activity, requiring a combination of dispositions, abilities and approaches that can be developed by drawing on a range of learning strategies. PMID- 9847714 TI - From classroom theory to clinical practice: evaluating the impact of a post registration course. AB - This paper is concerned with the translation of bioscience theory into clinical practice. The empirical data form part of an action research study undertaken to investigate the clinical outcomes of the bioscience component of a post registration, day release diploma in nursing taught in a higher education institution. Evidence for the impact of the course on patient care was collected by academic diary, questionnaire and interview. The triangulated data indicated that by adapting the pedagogic interpretation of the biosciences, it is possible to bridge the theory-practice gap. As a result, some nurses related the substance of the lessons to their patients, in order to bring about improvements in care. This paper examines the reasons behind the successes and failures of this continuing professional education initiative. PMID- 9847715 TI - The experience of facilitation in reflective groups: a phenomenological study. AB - The use of facilitation in all areas of nursing has increased in the last decade. There is little research related to the process of facilitation. In order to evaluate facilitation as an aspect of practice, an exploratory study was conducted to clarify the meaning of facilitation as used in practice. A phenomenological approach allowed identification of the meaning of facilitation from the lived experiences of practitioners who have implemented facilitation. Open interviews (conversations) were used to elicit relevant experiences from three nurse teachers who were engaged in the facilitation of reflective groups. The data collected were analysed using Collaizi's method of phenomenological data analysis. Having considered the findings of the study in the light of literature available related to facilitation, the implications for practice within nurse education are explored. PMID- 9847716 TI - Teaching nursing history: the Redwood experience. AB - This paper traces the development of nursing history from the great names approach of the early 20th century to the new analytical directions represented by the work of Davies and Maggs. It discusses the relevance of the study of nursing history in the present climate. The author shares the experiences of incorporating nursing history into a pre-registration diploma programme and suggests ways of evaluating this course of study. PMID- 9847717 TI - Shared understandings: negotiating the meanings of health via concept mapping. AB - This paper discusses a project undertaken to explore the effectiveness of concept mapping in assisting students to develop positive concepts of health. Positive concepts of health are central to the notion of health promotion. While strategies for health promotion are included in all nursing education programmes in Australia, to date there have been few strategies that are oriented towards promoting the development of positive concepts of health discussed in the literature. If student nurses are to become health-promoting practitioners, health teaching in nursing education must employ teaching strategies which clarify for students the differences between positive and negative concepts of health. One such strategy is concept mapping, which was developed as an educational tool to foster meaningful learning. Concept maps work to identify misconceptions in relation to a topic by clarifying the essential points, thus opening the way for concept change. This paper discusses the results of using concept mapping within the context of an abstract subject such as health, identifies difficulties in evaluating the strategy and makes recommendations about how these might be overcome. PMID- 9847718 TI - Freedom to enjoy learning in the 21st century: developing an active learning culture in nursing. AB - In 1994, the Wolfson School of Health Sciences was created in Thames Valley University, UK. This paper discusses an active learning approach being implemented throughout the university, and gives an outline of its application within nursing education. Practising as a confident, competent, reflective practitioner in the profession of nursing cannot automatically commence on the day of registration. Rather, the philosophical underpinnings necessary should be fostered throughout the initial nursing education curricula. In a political climate where there is a strong emphasis on efficiency, nurse teachers need to reconsider how the acquisition of nursing knowledge is facilitated. A description of how an active learning approach is being further developed in the nursing skills laboratory and the physiology laboratory is given. An outline identifies how these facilities may help students of nursing to gain knowledge and skills in relation to respiration. The authors recognize that while laboratories have been used widely in general education, their use has been limited in nursing education, particularly in the UK. Informal feedback from students implies that not only do they enjoy learning in these facilities, but also motivation towards learning is raised, an important element in the learning process. It is suggested that this will result ultimately in increased nursing competence. The authors acknowledge that these are preliminary informal findings and that formal investigation is required. PMID- 9847719 TI - Continuing education for registered nurses in the Highlands and Western Isles of Scotland. AB - This paper focuses on an educational approach used in the Highlands and Western Isles of Scotland in the continuing education of registered nurses. It includes a rationale for the educational approach used by this author and her colleagues, and a discussion of the approach itself. The discussion includes reference to concepts borrowed from learning theorists that underpin the approach, and case study material to illustrate how it works in practice. It is argued that the educational approach in this paper can potentially lead to meaningful learning. Meaningful learning is defined in this context. Finally, the outcomes of the educational approach are highlighted, with a discussion of its value to educators elsewhere. PMID- 9847720 TI - Writing ourselves: creating knowledge in a postmodern world. PMID- 9847721 TI - Flexibility: is it tying us in knots? PMID- 9847722 TI - Learning to nurse through legitimate peripheral participation. AB - Learning to nurse is identified as a complex interaction of affective, practical and cognitive factors. The nature and quality of the clinical learning environment is recognized as being influential in promoting the integration of them all. Students spend the majority of their time in a supernumerary capacity during their clinical placements and are dependent upon their mentor to engage them in increasingly sophisticated professional activities. Such activities should challenge students' understanding and promote their development of professional knowledge. In this longitudinal study, seven pre-registration nursing students were followed throughout the clinical experiences of their nursing degree programme. Questions were concerned with the nature of their experiences and the factors which influenced their professional development. These were pursued using documentary evidence supplied by the students, observations and regular focused interviews during each placement. Findings demonstrated the importance of a close and facilitative relationship which consequently enabled students to engage in activities contributing to their professional development. Working alongside a knowledgeable and respected practitioner led students to develop an enthusiasm and commitment to their professional development that was unparalleled. As a result, students were more likely to interact with other personnel working in the clinical environment and to become successful autonomous learners. Promoting their success was the nature of sponsorship afforded by mentors and the type of practice in which students were engaged. This has implications for mentorship preparation as well as the professional development of existing and future generations of nurses. PMID- 9847723 TI - Exploring the potential of problem-based learning in nurse education. AB - It could be argued that the role of the nurse is beginning to change from that of a medical assistant to one of a more research-aware, reflective professional. As a result, the education of nursing students has needed to adapt. This paper explores how problem-based learning (PBL) may be an appropriate means of achieving an educational preparation that can respond to changing needs. Particular attention is paid to the theoretical underpinnings of PBL, especially the concepts of adult education, cognitive processes and learning in context. Some of the relevant empirical studies are reviewed, and the authors attempt to identify the gaps in the literature, concluding by recommending a course of action to expand the existing levels of understanding and evidence that supports, or otherwise, the use of PBL. PMID- 9847724 TI - Problem-based learning: providing hope for psychiatric nursing? AB - Problem-based learning has been recognised as an educational method designed to facilitate student motivation and interest in clinically-based situations in order to increase their problem-solving capabilities. This paper describes the author's experience with the development and implementation of problem-based learning for the study of psychiatric nursing. The results of student evaluation suggest that this approach has been successful in fostering the interest of students in the skills and knowledge of this field of practice. More specifically, a stronger interest in pursuing psychiatric nursing as a career has emerged. This is particularly important in view of the negative image which the majority of undergraduate nursing students appear to hold towards psychiatric nursing as a career option. PMID- 9847725 TI - Current educational reforms in nursing in the United Kingdom and their impact on the role of nursing lecturers in practice: a case study approach. AB - This paper explores major developments in nurse education in the UK in the last 5 years and examines their impact on the role of nursing lecturers in practice. It builds upon the findings of an earlier study that described significant changes in the UK in the role and work of nurse teachers resulting from Project 2000 initiatives. Empirical data were collected over a 2.5-year period, through observations of student-teacher interaction and in-depth interviews with experienced nurse teachers and senior health professionals (n = 41). A case study approach using qualitative techniques was adapted for the study. The results of the study showed that, in the light of current educational reforms, nursing lecturers were expected to maintain closer links with the service sector and spend more time on clinical teaching and learner contact. Also considered important was the need for nursing lecturers to support clinical staff in creating an effective learning environment. Using the findings of the study, the author has mapped what was perceived to be the development of an effective model for the role of nursing lecturers in practice: a model that reflects the expected changes to a higher educational environment. PMID- 9847726 TI - Professional self-concept of nurses: a comparative study of four strata of nursing students in a Canadian university. AB - This study represents an effort to refine our understanding of measurement of professional self-concept by testing a tool across various strata of nursing students. The Professional Self Concept of Nurses Instrument, developed in studies of Australian nursing students, was applied to samples of basic undergraduate, post-basic and graduate students in a Canadian university nursing programme. The findings suggest that the tool is sensitive to the predictable development of professional self-concept as students advance through professional education, especially in the dimensions of professional practice and satisfaction. The authors propose that the instrument may have specific applications in evaluating the effects of traditional and innovative educational strategies upon the evolution of professional self-concept in nurses. PMID- 9847727 TI - Skills centre: why we did it, how we did it. AB - A review of current nurse education research in the UK, and relevant research from the USA has indicated that nursing students are experiencing difficulties in acquiring psychomotor skills. The reasons are possibly quite complex, but student and clinical staff evaluations point to short clinical placements and lack of college preparation as the cause. As a result of this review and a pilot survey, a skills centre has been built at Avon and Gloucestershire College of Health (Avon Site). This paper examines in detail the literature that led to the commissioning of the skills centre and provides practical guidelines on how a skills learning facility can be developed together with information on using a structured, but not confining, approach to teaching psychomotor skills. Further study will demonstrate how the skills centre can be evaluated using qualitative and quantitative research strategies. PMID- 9847728 TI - Educational clinical supervision: meeting the needs of specialist community practitioner students and professional practice. AB - In light of the possible demise of the community practice teacher, this paper proposes that a specific system of educationally led clinical supervision would benefit learning, teaching and assessing in the practice elements of specialist community practitioner education. Such a system would be additional to models of clinical supervision currently utilized in practice. An educative supervisory relationship would foster those skills and attributes essential for specialist community practice, enabling students to move toward mature responsible practice. It would provide experience of supportive frameworks that could be later utilized in professional practice. Clinical supervision specifically tailored for specialist community practitioner education would enable equity across branches and could be delivered in a variety of settings without a supervisor having to be 'on-site', thus benefiting comparatively isolated students in practice nursing and occupational health. To work successfully, educationally led clinical supervision necessitates a joint response by educational institutions and those practices where specialist community students undertake assessed placements. Without such a system in place to support specialist community practitioner courses, the standard of education and future practice cannot be assured. PMID- 9847729 TI - Students' knowledge of and attitudes towards primary health care. AB - The aims of this study were to assess nursing students' knowledge of and attitudes towards primary health care using a newly developed Canadian instrument, the Primary Health Care Questionnaire (PHCQ). The instrument was refined for use in the UK prior to collecting data from 427 students in degree, Project 2000, health visitor and district nursing courses. Findings indicate that students in all programmes surveyed have been exposed to the concepts of primary health care. Students have acquired knowledge about primary health care and, generally, positive attitudes to the concepts. Significant differences were found between several student groups on both the knowledge and attitude measures. Although further psychometric evaluation of this new measure is recommended, the PHCQ shows promise as a valuable measure for use in assessing the move toward an increased emphasis on primary health care concepts in nursing education. PMID- 9847730 TI - Application of the planning compass to the nursing curriculum: a tool for health promotion practice. PMID- 9847731 TI - The Internet: practical applications in nurse education [corrected]. AB - To a number of nurse educators, the Internet is a new, innovative and exciting teaching and learning tool. Sadly, anecdotal evidence suggests that many educators remain largely unaware of these new technologies or their potential to assist in the process of learning. Such a situation is not surprising given the limited amount of literature detailing the use of the Internet in nurse education and a profound lack of educational research in the area. This paper will highlight some of the benefits of the Internet for nursing students and educators, provide an overview of a number of Internet technologies and suggest some practical applications of these technologies in nurse education. A glossary of Internet terms is included. PMID- 9847732 TI - Course-related family and financial problems of mature nursing students. AB - In recent years there has been a significant change in the type of student entering nurse education. There has been a move towards widening access, and as a result, more mature students have gained entry to courses. What seems less clear is the types of course-related problems experienced by mature students. The study reported in this paper investigated the financial and family problems that mature students experience as a direct result of undertaking a course in nursing education. It was found that most mature students experience financial, childcare, relationship and family problems. It was also found that a large number of mature students thought about leaving the course as a result of these problems. It may be necessary, in the light of the findings, to consider how future course designs can accommodate mature students and their particular experiences. PMID- 9847733 TI - Learning clinical skills: the return of the practical room? PMID- 9847734 TI - Shared learning for community based maternity care. AB - In order to ensure the best quality of maternity care midwives and doctors must work closely together but unfortunately the necessary interdisciplinary cooperation and respect has not always been present. This paper suggests that the benefits claimed for interprofessional education, which remain largely untested in maternity care, are being put forward as a panacea for earlier difficulties. It outlines some of the initiatives currently taking place. Material ripe for shared learning is identified and it is suggested that shared teaching in addition to shared learning for midwifery and medical students and for midwives and doctors should be the norm. Learners may then eventually provide shared care based on both their shared learning experiences and also on the role model of shared teaching. Such a major change would require considerable political will, including a shift in funding. There is some evidence that the necessary political will is developing and this may be given more emphasis by the need for teaching to follow care as the NHS becomes more primary-care led. PMID- 9847735 TI - Devolution or centralization? Differences in the development of nurse education commissioning policy among the UK nations. AB - An overview of the different systems for the commissioning of nurse education in the various UK countries is contextualized in the policy concerns of development and marketization driven by Government Working Papers 9 and 10. The different systems for commissioning nurse education in the various UK countries are described, with a particular focus on the English consortia system and its level of efficiency. It is argued that while the mechanism of consortia is being used to marketize nurse education commissioning in England, in the other three nations the commissioning process has actually become more centralized. This centralization is contrary to the policy of development, yet it is suggested here that the consortia mechanism adopted in England has not been followed in the other UK nations, as it has not yet proved an independently efficient method of commissioning. PMID- 9847736 TI - Evaluating a skills centre: the acquisition of psychomotor skills in nursing--a review of the literature. AB - In 1994, as a result of both programme evaluations which identified students' fears and apprehensions about their practical ability, and a review of the literature on skill acquisition, experiential skills teaching was resumed within the faculty. Having invested considerable finance into the reconstruction of a skills centre to teach skills, it is now imperative that its use be formally evaluated. Part of the evaluative process includes a review of the empirical literature on the acquisition of psychomotor skills in nursing. This paper summarizes this review. PMID- 9847737 TI - Evaluating a skills centre: learning psychomotor skills--a review of the theory. AB - The formal teaching of skills based on the theoretical principles of skill acquisition has, so far, been practised by teachers responsible for preparing students for their first adult clinical placement on the diploma programme. Whether teachers on other programmes are using similar models of skill acquisition has been surveyed, and the results are presently being analysed. This approach to teaching skills within the context of the Skills Centre is also being evaluated. However, as indicated in the introduction to this paper, it is important to develop and implement the theoretical foundations for skills teaching before any evaluative research on it can take place (Chen 1986). Now these foundations are in place, the evaluative strategy can proceed. PMID- 9847738 TI - Learning clinical skills: an interprofessional approach. AB - The professions of nursing and medicine are committed to interprofessional education, in the belief that through this, patient care and satisfaction will be improved. Most initiatives involving nurses have been at post-qualification level, in primary health care, and concerning interpersonal or information management skills. Much of this collaboration has been with professions allied to medicine or social services. This paper discusses an innovative programme of shared learning in acute care, involving final year medical students and newly qualified staff nurses. The programme, developed in response to the blurring of professional roles between nurses and junior doctors, took place in our interprofessional Clinical Skills Centre. It was based around a developing patient scenario which was pertinent to the participants' area of practice. Each session was led by an experienced nurse lecturer and doctor, supported by specialist contributors. The style of learning was participative, with small interprofessional groups addressing a range of patient management issues. In this way, relevant clinical and communication skills were integrated within the context of holistic patient care. The course was well evaluated by both professional groups of participants and their managers. Subsequent research and curriculum development are leading to the expansion of this successful initiative. PMID- 9847739 TI - The nature of the problem: the intentional design of problems to facilitate different levels of student learning. AB - This paper explores the deliberate design of 'problem' situations within a 3-year problem-based nursing degree conducted at Griffith University, Nathan, Queensland, Australia. It examines problem situations within the first and third years of the course to illustrate the issues involved in designing material that enables students to achieve the curriculum aims. Although the problem situations are designed from the same health care setting, the discussion highlights how concepts and issues can be addressed in different ways depending on the desired student outcomes. The paper outlines and examines the nature of student learning and the clinical reasoning process used by students in making decisions that closely mirror those made as registered nurses in order to facilitate the achievement of clients' health goals. PMID- 9847740 TI - Working in syndicate groups towards the development of clinical care protocols: a study into the professional learning of undergraduate nursing students. AB - Group work is an important element in nurse education and practice, and is an issue of consistent relevance given the increasing emphasis on collaborative and multidisciplinary working, and the fact that most nurses' work involves them in a variety of groups. This study is ongoing research, and considers the experiences of five groups, each comprising four final year nursing undergraduate students working in syndicate groups. The students are working towards the creation and development of clinical care protocols. The study method is grounded theory. Data have been collected over 3 academic years, with the group facilitator as researcher. The data management involves tape recording meetings and transcribing these, with continual reduction and analysis of the data. Findings from early analysis suggest that important phenomena, for example 'resistance', 'tangible action goals', and the concept of 'closure', play a key part in the way in which the groups under study have functioned. PMID- 9847741 TI - Alcohol-related problems: a critical review of the literature and directions in nurse education. AB - It is generally accepted the around 2-5% of the adult population show major signs of alcohol dependence, that alcohol-related harm is experienced by up to 20% of the population, and that approximately 60% drink at risk-free levels. Further prevalence studies show that there are high numbers of problem drinkers who attend general hospital services for reasons other than their alcohol consumption. Nurses are in constant contact with patients who may have an early problem with alcohol but who are admitted for other reasons, and they are in a prime position to comprehensively assess patients (including alcohol screening), develop rapport and provide 'counselling'. Also, university nursing education is propelling nurses toward adoption of independent discipline focused models of care which are increasingly becoming independent of the medical model. Recent trends in the management of problem drinkers suggest that controlled drinking approaches may well offer treatment options to nurses that the traditional abstinence approaches did not. This paper presents a brief overview of the notion of controlled drinking, then critically reviews the nursing research studies and the descriptive literature providing direction for nursing education. Some recent clinical initiatives are discussed which highlight the flaws existing in nursing education, including lack of sufficient curriculum hours and the need for better designed education models and strategies. PMID- 9847742 TI - Applying the Mims-Swenson sexual health model to nurse education: offering an alternative focus on sexuality and health care. AB - Several recent publications have documented the existence of negative attitudes towards lesbians, gay men and bisexual men and women among health care professionals. It has been demonstrated that these negative attitudes predate the advent of HIV/AIDS, and are ingrained in medical and nursing education. Research has shown that, within the hospital environment, negative attitudes towards homosexuality may be directly translated into the medical and nursing care provided for lesbian, gay and bisexual clients. In this article, studies focusing on the attitudes of health care professionals towards homosexuality and bisexuality are examined, and their implications are discussed with reference to the future of nursing education. PMID- 9847744 TI - An evaluative study into the effectiveness of level 3 National Vocational Qualification support staff to nurses. AB - A trial development of experienced nursing auxiliaries using the level 3 National Vocational Qualification (NVQ) awards in care commenced in 1993 as part of an evaluative study to consider the implications and potential benefits of this initiative for an acute National Health Service Trust. Two small cohorts (n = 17) undertook development programmes. With the first candidates gaining the award in 1995, an evaluative study was undertaken to assess the quality of care delivered by these staff. Using a modification of the Qualpac assessment tool, trained observers compared the quality of care delivered by the NVQ workers with that delivered by different grades of nurses in three specialist areas: medicine, general surgery and elderly care. The findings reveal that the NVQ-prepared staff performed considerably better than conventional nursing auxiliaries, and in many cases, outperformed qualified nurses in terms of quality of care. The study compares results across a range of nursing skills and discusses the possible implications. PMID- 9847743 TI - Practice placement supervision: the role of the personal tutor. AB - Confusion regarding the role of the nurse teacher in clinical practice has resulted in a service being offered to students that varies in type, quantity and quality as perceived by students, teachers and placement staff alike: The role of the personal tutor is concerned with helping the students to develop both professionally and academically. To facilitate this, a good interpersonal relationship between student and personal tutor is essential. As interpersonal relationships take time to develop, the author advocates capitalizing on the relationship that (usually) already exists between personal tutors and students, and extending this into practice placement areas. The aim of this piece of research was to ascertain the value, as perceived by the students, of having the personal tutor supervise them in college as well as in the four branch placements of foundation studies. The methodology employed was a descriptive study utilizing a self-administered questionnaire. The population was the students in foundation studies of the Lothian College of Health Studies, and the sample was one of convenience, i.e. one cohort of students. The findings demonstrated that 55 students (n = 94) perceived value in having one nurse teacher supervising them in college and in placements compared with 39 students who did not perceive any value. The students were asked about their practice placement supervision by nurse teachers in terms of type, quantity and quality of teaching. The students in the former group rated their practice placement supervision much lower than students in the latter group. PMID- 9847745 TI - Racial discrimination in nursing. PMID- 9847746 TI - Shining through. PMID- 9847747 TI - Fighting back. PMID- 9847748 TI - A watch on staff welfare. PMID- 9847749 TI - A quick fix will not do. PMID- 9847750 TI - Wrong road. PMID- 9847751 TI - Heart to heart. Interview by Julie McCaughan. PMID- 9847752 TI - Skilled care counts. PMID- 9847753 TI - Baby love. Interview by Charlotte Alderman. PMID- 9847754 TI - Water world. PMID- 9847755 TI - Ethical and legal aspects of nutrition and hydration. AB - There is increasing realisation that malnutrition is a complication of many illnesses, as well as a consequence of inadequate food intake. The report summarised below, provides recommendations and principles to guide health professionals facing dilemmas concerning patients' nutrition and hydration. PMID- 9847756 TI - Sexual health for people with learning disabilities. AB - This article is based on work into the sexual health needs of people with learning disabilities, carried out by a working group of the RCN Family Planning Forum. Literature searches were conducted, and local initiatives, specialist services and information sources for training and education for clients and staff were identified. The authors' aim in this article is to increase awareness among those involved in the care of people with learning disabilities and those providing family planning and sexual health services, so that these services are appropriate, accessible and sensitive to the needs of this vulnerable client group. PMID- 9847757 TI - Implementing change: influencing the process. AB - In last week's Nursing Standard (Callaghan 1998) we reported the findings of a study into the effectiveness of plastic aprons in preventing contamination of nurses' uniforms. As a result of the study, the author made a series of recommendations to the infection control committee in her area. Although some of these recommendations were acted upon, the major changes recommended were not implemented despite evidence from the study. In this article, the author summarises the findings and recommendations of the study, and examines potential reasons why major changes were not carried out. PMID- 9847758 TI - The iatrogenic epidemic. AB - Medicine can often cause unnecessary harm to people while trying to treat them, but this phenomenon, known as iatrogenesis, is rarely debated. The author of this article argues that nurses have a responsibility to consider the concept of iatrogenesis and its effects on the health of their patients. PMID- 9847759 TI - Tracheostomy: suctioning and humidification. AB - This article explains tracheostomy suctioning and humidification. It outlines how nurses can assess the need for tracheostomy suctioning to be performed and possible complications that may occur as a result of the procedure. PMID- 9847760 TI - Pay, recruitment and conditions of service for nurses continue to hit the headlines. PMID- 9847761 TI - Creative recruitment. PMID- 9847762 TI - Find out what's going on. Interview by Kath Burke. PMID- 9847763 TI - Top up gets thumbs down. PMID- 9847765 TI - Calm down. PMID- 9847764 TI - Keeping in touch. PMID- 9847766 TI - Everyone's evidence. PMID- 9847767 TI - Nurses off the peg. PMID- 9847768 TI - Homing instinct. Interview by Charlotte Alderman. PMID- 9847769 TI - Referencing the Internet. PMID- 9847770 TI - Laying aside. PMID- 9847771 TI - Social systems intervention. AB - North and south Birmingham mental health trusts are piloting the application of social systems approaches to care with some of their community mental health teams. In this report, the author describes why social systems intervention (SSI) is an approach worth pursuing. PMID- 9847772 TI - Bacterial contamination of nurses' uniforms: a study. AB - This study examined the hypothesis that the wearing of plastic aprons during direct patient contact would reduce significantly the number of bacteria carried on nurses' uniforms, and therefore reduce the probability of the transmission of nosocomial infections. Current nursing practices and overall bacterial uniform contamination levels were investigated, as well as the effects of the wearing of plastic aprons to protect uniforms. The conclusions of the study demonstrate that such contamination may be significant contributory factor in the spread of nosocomial infections, and have implications not only for the nursing profession, but also for other members of the multidisciplinary team. In next week's Nursing Standard, the author examines the difficulties of putting research into practice. PMID- 9847773 TI - Streamlining pre-operative assessment in orthopaedics. AB - The author was appointed to create a 'pre-admission clerking system' in an orthopaedic department. The management objective was to reduce the numbers of patients whose surgery was cancelled. In this article the author describes the holistic assessment which has resulted in fewer operations being cancelled and reduced length of stay. PMID- 9847774 TI - Infection control: principles for practice. AB - This article discusses the general principles of infection control and outlines the role nurses can play in maintaining a safe environment. This article is the third and final part of our 1998 series on infection control. PMID- 9847775 TI - Nurse consultant posts with the same status in nursing as consultants enjoy in medicine. PMID- 9847776 TI - Yellow card for violent patients. PMID- 9847778 TI - Carry on nursing. PMID- 9847777 TI - Global warning. PMID- 9847779 TI - More than money. PMID- 9847780 TI - Getting what you pay for. PMID- 9847781 TI - The three Rs. PMID- 9847782 TI - Grim reality. PMID- 9847783 TI - Quality time. PMID- 9847784 TI - Sell your skills. PMID- 9847785 TI - Men behaving well. Interview by Charlotte Alderman. PMID- 9847786 TI - One stop to the best patient care. Interview by Charlotte Alderman. PMID- 9847787 TI - Links with the law. PMID- 9847788 TI - Lonely planet. PMID- 9847789 TI - After all the rhetoric.... PMID- 9847790 TI - Orthopaedic audit. PMID- 9847791 TI - Continuity of care for older people. AB - Older people transferring from NHS hospitals to independent sector nursing homes need continuity of care. However, nurses in the NHS and independent sectors are often out of touch with each other. In the second of two articles on discharge and older people, the author discusses the need for education, assessment and effective communication to help nurses, wherever they work, to provide effective care for elderly people. The author describes an assessment scale for physical and mental abilities, developed to assist the transfer process. The first article was published in Nursing Standard (Nazarko 1998). PMID- 9847792 TI - Grounded theory: issues for research in nursing. AB - The grounded theory approach to research appears to be well suited to providing nurses with understanding of social behaviours so that they can enhance patient care Here, the author examines the principles of grounded theory and describes how the approach can influence the way that nurses practise. PMID- 9847793 TI - Assisted ventilation in cystic fibrosis: nursing care. AB - In this article the author explains nasal intermittent positive pressure ventilation (NIPPV) for patients with cystic fibrosis (CF). Indications for use are noted before nursing care of the patient is described in detail. PMID- 9847794 TI - Decontamination of equipment and the environment. AB - This article explains the various ways decontamination of equipment and the environment can be carried out and highlights the importance of nurses being familiar with such methods so to reduce the possibility of infection. PMID- 9847795 TI - Capital opportunities. PMID- 9847796 TI - Support for nurses to be rewarded properly for their work. PMID- 9847797 TI - Pay: time is running out. PMID- 9847798 TI - Dramatic developments in nursing. PMID- 9847799 TI - No place to hide. PMID- 9847800 TI - Missing link. PMID- 9847801 TI - Health waves. PMID- 9847802 TI - All tired out. PMID- 9847803 TI - Home and away. PMID- 9847804 TI - Red card for allergies. Interview by Charlotte Alderman. PMID- 9847805 TI - Busy doing ... nothing much. PMID- 9847806 TI - More pennies in the bank. PMID- 9847807 TI - Hospital-acquired infection. AB - This report focuses on nosocomial or hospital-acquired infections (HAIs), discussed extensively at this year's annual conference of the Infection Control Nurses' Association (ICNA) in September. PMID- 9847808 TI - Effects of stress in an NHS trust: a study. AB - This study of staff in an NHS trust tests two established models of occupational stress. Payne (1979) suggested that support at work can help to neutralise the strain of work demands, while Karasek (1979) suggested that staff whose jobs are characterised by high demands and low control are at greater risk of poor psychological wellbeing and ill health. Support is found for both models, and it is suggested that the two could usefully be combined. PMID- 9847809 TI - Biological hazards. AB - This article outlines the biological hazards that nurses may face in their workplace. It hopes to inform nurses how they can reduce such hazards at work and contribute to a safer working environment. PMID- 9847810 TI - The importance of nutrition in wound healing. AB - Nursing Standards recent Focus on Nutrition campaign raised some uncomfortable issues about nurses' knowledge of wound healing. We continue the theme here, with a review of the importance of good nutrition for effective wound healing. PMID- 9847811 TI - Repositioning to prevent pressure sores--what is the evidence? AB - The prevention of pressure sores by frequent repositioning of patients is a widely accepted practice. In this article, the author questions whether this should be so and suggests that there may be more effective ways of relieving pressure damage. PMID- 9847812 TI - Tissue donation: the benefit of a positive approach. AB - Unlike organ donation, which enjoys a worthy public profile, tissue transplantation remains relatively unheard of. Here, the authors argue that nurses should promote the value of tissue donation, not only to patients but to colleagues as well. PMID- 9847813 TI - Dreaming spires. PMID- 9847815 TI - Transitioning to independence: challenges for young people with disabilities and their caregivers. AB - For all teens, the process of moving from childhood to adulthood is challenging. For young people with disabilities, transitioning to independence presents even more challenges. Barriers to successful transition for young people with disabilities include low expectations by parents and other significant people in the community, lack of knowledge of existing career and vocational education services, and lack of self-advocacy skills. This article provides an overview of issues related to transitioning to adult independence and offers suggestions for assessment, planning, and intervention that can help nurses be effective partners with families and other caregivers in transition efforts. Nurses caring for children with disabilities can help families see strengths in their children and develop realistic, developmentally-appropriate expectations for skill development, attitudes, and behaviors that will promote self-sufficiency in adulthood. Nurses can help families think about possibilities for independence and refer families to community resources that can help young people with disabilities pursue postsecondary education, obtain and maintain jobs, and live independently. PMID- 9847814 TI - Prevention of hip fractures: a nursing imperative. AB - Hip fractures are one of the leading causes of morbidity and mortality among the elderly, and they greatly impact health care costs. Most hip fractures in the elderly are due to osteoporosis. This paper reports the incidence of hip fracture, describes osteoporosis, and discusses the role of bone mass in the development of this disorder. Risk factors for osteoporosis and, specifically osteoporotic hip fracture, are identified. Preventive strategies for osteoporotic hip fracture, including fall prevention, are discussed in detail, and pharmacologic interventions are described. Research evidence indicates that the prevention of hip fracture is realistic, and most of the intervention strategies fall directly in the realm of nursing. PMID- 9847816 TI - Management of Colles' fractures. AB - The Colles' fracture is probably the most common fracture seen in the Emergency Department. Although there are several different methods of reduction and fixation, the goal of any treatment plan is to return the patient to normal function. Depending on the complexity, the Colles' fracture can be successfully treated open or closed. This article reviews the mechanism of injury, assessment, conservative or operative treatment, and rehabilitation of these fractures. PMID- 9847817 TI - Using conflict to improve effectiveness of nurse teams. AB - As nurses increasingly turn to teamwork as a viable option for accomplishing a myriad of duties and responsibilities, they are discovering that teamwork can also be a source of conflict. This article suggests that conflict is both inevitable and necessary within a team. A model is presented that aims not at reducing conflict but at making it more effective. The author traces the influence of preconflict conditions and perspective-taking on conflict and ultimately team effectiveness. Finally, to aid in applying the model to practice, two examples are presented, demonstrating both an ineffective and an effective use of conflict. PMID- 9847818 TI - Implementing a bicycle safety workshop. AB - Each year almost 400,000 children ages 14 and younger are treated in emergency rooms for bicycle-related injuries. An additional 500,000 young riders are treated in other settings (private pediatrician's office, urgent care centers) (American Academy of Pediatrics, 1991). An awareness has been raised this past decade toward programs that target injury prevention and promote safety and health in children. The implementation of bicycle safety workshops and "bicycle rodeos" are just one of many programs that educate young children on the techniques of safe, yet fun, bicycling. PMID- 9847819 TI - Osteoporosis prevention in young women. AB - PURPOSE: To assess whether young women who participate in an osteoporosis prevention program based on the Health Belief (Rosenstock, 1966) and Self Efficacy Models (Bandura, 1977) demonstrate higher levels of knowledge regarding osteoporosis prevention than young women who do not participate in such a program. DESIGN: A classic experimental design with one treatment group and one control group was used to test the efficacy of the osteoporosis prevention program. Pretest data on knowledge, health belief attitudes (7 subscales), and self-efficacy (2 subscales) were collected in the treatment and control groups. SAMPLE: A convenience sample of 31 young college women were randomly assigned to an experimental group or to a control group to receive an osteoporosis prevention program. METHOD: Subjects in the experimental and control groups completed the Osteoporosis Knowledge Test, the Osteoporosis Health Belief Scale, and the Osteoporosis Self-Efficacy Scale (Kim et al., 1991) at two times. The experimental group received an osteoporosis prevention program. MAIN RESEARCH CLASSIFICATIONS: Osteoporosis, Health Belief, Self-Efficacy, Women's Health. FINDINGS: Subjects in the experimental group had significantly higher knowledge and health belief scores after receiving the intervention than their pretest scores while subjects in the control group had no change in scores. CONCLUSION: The osteoporosis program was effective in increasing awareness of osteoporosis prevention in this group of young women. IMPLICATIONS FOR NURSING RESEARCH: The results may be useful for developing young women's awareness and knowledge of osteoporosis prevention. Future research could include developing osteoporosis prevention programs at an earlier age when girls are in grade school and junior high school. Teaching health promotion strategies for bone health is essential to all women across the life span. PMID- 9847820 TI - Treatment of low back pain with intraspinal opioids delivered via implanted pumps. AB - Low back pain, more recently defined as "failed back surgery syndrome" is a common, yet devastating, pain complaint. Although physical therapy and systemic analgesics provide relief in most patients, intraspinal opioids delivered via implanted pumps may be indicated in those individuals who cannot tolerate oral medications. Orthopaedic nurses are essential in the identification and care of patients who might benefit from this therapy. PMID- 9847821 TI - Developing continuing education for contact hour approval: an update. AB - Developing quality nursing continuing education for contact hour approval requires a basic understanding of the nursing continuing education system and the application process. The Approver Unit of the National Association of Orthopaedic Nurses (NAON) helps providers (i.e., members, chapters, corporations) develop educational activities and grants contact hour approval. This article provides the latest information on nursing continuing education terminology and the application process for contact hour approval through NAON. PMID- 9847822 TI - Building clinical pathways. AB - TQM principles change the work environment so that point-of-service personnel can improve health care delivery to patients. The clinical pathway process starts with the principles of TQM. In the era of managed care, health care resources can be managed effectively using a clinical pathway. The multidisciplinary team has the opportunity to improve the health care services provided to patients. PMID- 9847823 TI - Managed care of outpatient psychotherapy: a new twist. PMID- 9847824 TI - Paradox as symptom in the borderline patient's struggle for self-differentiation. AB - TOPIC: Paradox as symptom in patients diagnosed with borderline personality disorder (BPD). PURPOSE: To conceptualize the nature of difficulties arising in psychotherapy with patients diagnosed with BPD. SOURCES: Review of the literature and clinical data from the author's work. CONCLUSIONS: The borderline dilemma is inherently paradoxical and, in some ways, reflects the human condition. Psychoanalytic concepts can further our understanding of the therapeutic difficulties encountered in the struggle to make sense of life's inherent dualities. The borderline patient is acutely sensitive to these dualities, which the therapist must tolerate and understand. Often, a negative therapeutic reaction may impede progress. PMID- 9847825 TI - A multidisciplinary course to teach staff to conduct psychodynamic group psychotherapy for assaultive men. AB - TOPIC: A course in leading psychodynamic psychotherapy groups for assaultive men. PURPOSE: To teach multidisciplinary staff members to lead these groups, using both didactic and experiential learning. SOURCE: The author's own experience. CONCLUSIONS: Multidisciplinary staff members learned about the process of leading psychotherapy groups for assaultive men by attending lectures and observing the author lead a group over a 6-month period. The author's openness about her own thoughts, feelings, fantasies, and countertransference allowed students to explore their own reactions and to empathize with the patients, observing that they were not so different from them. PMID- 9847826 TI - Factors influencing movement of chronic psychiatric patients from the orientation to the working phase of the nurse-client relationship on an inpatient unit. AB - PROBLEM: To identify factors influencing movement of nurse-patient dyads from Peplau's orientation phase to the working phase of the nurse-client relationship in a tertiary care psychiatric setting. METHODS: Ten nurse-client dyads were interviewed after the initial nurse-client assignment until a consensus was reached between client, nurse, and investigator/CNS that the relationship was in the working phase. FINDINGS: Factors causing the relationship to progress, from the clients' perspective, were the perceived attitude of the nurse, the nature of the planned therapeutic sessions, and what happened between therapeutic sessions. Factors hindering the development of the relationship included a nurse's or client's unavailability, a sense of distance/inequity, differences in realities/values, and mutual withdrawal. The relationship was perceived as supportive and "powerful" when it progressed to the working phase, but as very negative and like "limbo" if instead it moved to a phase of mutual withdrawal. CONCLUSIONS: Nurses can help clients move from the orientation phase to the working phase by remaining available, consistent, and acting in a way that promotes trust. When the relationship does not progress to the working phase within 6 months, a therapeutic transfer should be considered. PMID- 9847827 TI - First annual nurse psychotherapist conference: mind over brain. I. PMID- 9847828 TI - At last, Medicare reimbursement. PMID- 9847829 TI - The reification of the "pathological" gambler: an analysis of gambling treatment and the application of the medical model to problem gambling. AB - PROBLEM: Is the medical model used to reify "pathological" gambling? METHODS: A qualitative field study of 12 subjects with the diagnosis "pathological" gambler. FINDINGS: None of the 12 subjects being treated for "pathological" gambling fit neatly in the category of "pathological" gambler. An examination of the treatment center revealed institutional grounds for the use of this diagnosis. CONCLUSIONS: Rather than a medical diagnosis, "pathological" gambling is a socially constructed phenomenon. Using the medical model serves clients interpersonally and clinicians occupationally. PMID- 9847830 TI - Protecting partial-hospitalization patients from suicide. AB - TOPIC: Assessing suicide potential and intervening to prevent suicide in partial hospitalization patients. PURPOSE: To provide information for assessing and intervening with outpatients who express suicidal thoughts and to propose a decision tree. SOURCE: Literature review, clinical experience. CONCLUSIONS: Careful assessment and sound management are required to provide treatment in outpatient settings while maintaining patient safety. A decision tree can help the clinician make sound decisions. PMID- 9847831 TI - Factors that contribute to satisfaction or dissatisfaction in stepfather stepchild relationships. AB - TOPIC: Factors contributing to satisfaction and dissatisfaction in stepfather stepchild relationships. METHOD: A qualitative multicase study of six stepfathers. FINDINGS: Four themes emerged: previous experiences as parents, other relationships within the family, issues that required negotiating, discipline of the children. CONCLUSIONS: Previous experience as parents, good relationships with other family members, similarity in values between parents, and inclusion in the disciplinary process increased stepfathers' satisfaction. PMID- 9847832 TI - Hold applause for PL 105-33. PMID- 9847833 TI - Children killing children. PMID- 9847834 TI - Self-resolution of drinking problems as a process of reinvesting in self. AB - PROBLEM: Resolution of alcohol problems without formal treatment or participation in self-help groups. METHODS: Qualitative study using grounded theory (N = 11). FINDINGS: The onset of alcohol problems begins with negligible penalties. Over time, the cost-benefit ratio of drinking habits continues to rise and the risks become too great. Individuals find it necessary to change their drinking patterns by reinvesting in themselves. Assets such as the ongoing availability of information, life-management skills, and self-confidence promote the change process; cultural mores and behaviors of some healthcare providers serve as liabilities. The dividends of self-resolving alcohol problems include self-pride, mental and physical health, conscientious work performance, rewarding relationships, enhancement of creative talents, and spiritual well-being. CONCLUSIONS: Nurses can play an important role in promoting self-resolution of alcohol problems by providing accurate information and encouraging clients to reinvest in long-standing priorities and values. PMID- 9847835 TI - Strategies for maintaining optimal wellness in the chronic mentally ill. AB - TOPIC: Ways the chronic mentally ill avoid loss of control and the importance of both the therapeutic environment and the nurse-patient relationship in helping them do this. PURPOSE: To examine the strategies people with chronic mental illness use to prevent loss of control and maintain health, and ways nurses can help with this process. SOURCE: Findings from a previously published study of seven chronically mentally ill clients in a rural day-treatment center. CONCLUSIONS: Four properties related to the core variable of "preventing loss of control" are relationships, feelings, good attitude, and functional activity. These properties must be examined within the context of the therapeutic environment as well as the nurse-patient relationship. PMID- 9847836 TI - Using Margaret A. Newman's theory of health with elderly nursing home residents. AB - TOPIC: The use of Margaret A. Newman's theory by psychiatric nursing students with elderly nursing home residents. SOURCE: The author's work with baccalaureate nursing students. CONCLUSIONS: Nurses who form relationships with elderly nursing home residents can help their clients identify problematic patterns, move to a choice point, then to a higher level of consciousness, resulting in a reduction in problematic patterns of behavior. PMID- 9847838 TI - [Four paws for ... two legs]. PMID- 9847837 TI - Reconstructing a sense of self in patients with chronic mental illness. AB - TOPIC: The loss and reconstruction of self in patients with a chronic mental illness. PURPOSE: To describe the loss of self and its reconstruction. SOURCES: The authors' own clinical work and review of the literature. CONCLUSIONS: Nurses can help patients who have undergone a loss of self to discover a more active sense of self, take stock of the self, put the self into action, and use the enhanced self as a refuge. This process involves the fostering of hope. PMID- 9847839 TI - [Four paws for ... two eyes]. PMID- 9847840 TI - [Not to add anxiety to pain]. PMID- 9847841 TI - [Hidden distress: the endangered "elderly". Does institutional violence exist? What is its nature?]. PMID- 9847842 TI - [Selection of workshops for the therapeutic stimulation of the elderly]. PMID- 9847843 TI - [Adding stimulation and rhythm to everyday life in an institution]. PMID- 9847844 TI - [Security systems helping the elderly at home]. PMID- 9847845 TI - [Virginia Henderson's "14 fundamental necessities". Interview by O Burrus]. PMID- 9847846 TI - [A current problem: health safety]. PMID- 9847847 TI - [Psychological problems and hospitalization: recognizing in order to treat better]. PMID- 9847848 TI - [The American health care system: nurses of honor]. PMID- 9847849 TI - [Open letter to a "psychiatric patient"]. PMID- 9847850 TI - [The Permanent Committee of Nurses of the European Union]. PMID- 9847851 TI - [To be a European citizen and a professional in today's society: what does it mean?]. PMID- 9847852 TI - [Nurses in this hour of Europe]. PMID- 9847853 TI - [Nurses, research and Europe]. PMID- 9847854 TI - [Positions of nurses within the World Health Organization in Europe]. PMID- 9847855 TI - [The creation of health care frameworks in Europe]. PMID- 9847856 TI - [What kind of health care establishment for the Europe of the 3. millennium?]. PMID- 9847857 TI - [The function of organizing at the beginning of the 21. century]. PMID- 9847858 TI - [Opening up of hospitals all over Europe]. PMID- 9847859 TI - [FINE (European Federation of Nurse Educators)]. PMID- 9847860 TI - [New training within the French community in Belgium]. PMID- 9847861 TI - [The post-nursing diploma education in Great Britain and especially in Scotland]. PMID- 9847862 TI - [The 21st century, what are the challenges for nursing?]. PMID- 9847863 TI - [The lifting of patients. An optional model]. PMID- 9847864 TI - [The helping relationship. Collaboration between the nurses of a palliative care mobile unit and the nurses of the clinical services]. PMID- 9847865 TI - [Continuing education: the return]. PMID- 9847866 TI - [Don't let that opportunity pass you by. Interview by Anne Boiteux and Bernard Goddet]. PMID- 9847867 TI - [Miss Rosette, nurse among the Haitian peasants. Interview by Aurore Le Maitre]. PMID- 9847868 TI - [Building our cities in the mountains...]. PMID- 9847869 TI - [Epidemiology of asthma in children]. PMID- 9847870 TI - [Treating an asthmatic crisis at the hospital]. PMID- 9847871 TI - [The basis for the treatment of asthma in children]. PMID- 9847872 TI - [Education of the asthmatic child]. PMID- 9847873 TI - [The place of kinesitherapy]. PMID- 9847874 TI - [The benefits of climate treatments]. PMID- 9847876 TI - [Oral and dental hygiene]. PMID- 9847875 TI - [Prevention of pain in the premature. Electroencephalographic recording]. PMID- 9847877 TI - [Organ donation]. PMID- 9847878 TI - The future of clinical dentistry. PMID- 9847879 TI - Risk assessment in dentistry. PMID- 9847880 TI - Risk assessment and caries prediction. PMID- 9847881 TI - Caries diagnosis. PMID- 9847882 TI - Restorative dentistry: current status and future directions. PMID- 9847883 TI - Management of dental caries as a chronic infectious disease. PMID- 9847884 TI - Caries prevention. AB - It is apparent that the use of fluoride in multiple measures has a significant impact upon the prevention of dental caries. These measures involve public health benefits of water fluoridation, professional fluoride treatments in the dental office, and the home use of effective fluoridated dentifrices, with the use of fluoride rinses and gels as adjuncts when needed. In many clinical situations, professional judgment is required to identify the most appropriate treatment measures to address the needs of individual patients. PMID- 9847885 TI - Periodontal diseases: a new paradigm. PMID- 9847886 TI - Risk assessments in the diagnosis and treatment of periodontal diseases. PMID- 9847887 TI - Inheritance as a determinant of susceptibility for periodontitis. PMID- 9847888 TI - Role of periodontitis in systemic health: spontaneous preterm birth. PMID- 9847889 TI - Oral health and systemic disease: periodontitis and cardiovascular disease. PMID- 9847890 TI - Periodontal disease diagnosis and treatment: an exciting future. PMID- 9847891 TI - Impediments to change and their resolution in clinical practice. PMID- 9847892 TI - Something particular in the air we breathe? PMID- 9847893 TI - What is the truth about surgical waiting lists? PMID- 9847894 TI - Female genital mutilation: responding to health needs. PMID- 9847895 TI - "You've gotta have HAART". PMID- 9847896 TI - Outdoor air pollution and children's respiratory symptoms in the steel cities of New South Wales. AB - OBJECTIVE: To investigate the relationship between outdoor air pollution and the respiratory health of children aged 8 to 10 years. DESIGN: A cross-sectional survey (between October 1993 and December 1993) of children's health and home environment. Summary measures of particulate pollution (levels of particles with an aerodynamic diameter less than 10 microns [PM10] each 6th day) and SO2 (daily mean and maximum hourly values) were estimated for each area (using air quality monitoring station data from July 1993 to June 1994). SETTING AND SURVEY PARTICIPANTS: Parents of 3023 primary school children (Years 3, 4 and 5) from industrial and non-industrial areas with air quality monitoring stations in the Hunter and Illawarra regions of New South Wales. MAIN OUTCOME MEASURES: Reported occurrence of four or more chest colds, four or more attacks of wheezing, and night-time cough without a cold for more than two weeks, all within the previous 12 months. RESULTS: 77% response rate, ranging by area from 66% to 88%. The average annual outdoor air pollution for the nine areas was 18.6-43.7 micrograms/m3 for PM10 and 0.16-0.90 parts per hundred million for SO2. The proportion of children reported to have the main outcome symptoms were: chest colds, 3.0%-9.7%; night cough, 12.3%-30.5%; and wheeze, 3.4%-11.3%. There was no significant association with SO2, but a significant increase in the odds of symptoms per 10 micrograms/m3 increase in PM10 on chest colds (odds ratio [OR], 1.43; 95% confidence interval [CI], 1.12-1.82) and night-time cough (OR, 1.34; 95% CI, 1.19-1.53), but not wheeze. Passive smoking was significantly associated with chest colds but not with the other symptoms. Maternal allergy was associated with all three respiratory symptoms, most strongly with wheeze. CONCLUSION: These results provide evidence of health effects at lower than expected levels of outdoor air pollution in the Australian setting. They also suggest differences in contributions of environmental and hereditary factors to cough and chest colds compared with wheeze. PMID- 9847897 TI - Factors associated with waiting time for surgery. AB - OBJECTIVE: To explore the factors associated with waiting times for surgery in public hospitals. SETTING: Three major acute care public hospitals in one Area Health Service. PARTICIPANTS: 691 adult patients entered onto the computerised booking list between 16 and 22 November 1994, and then between 16 January and 12 February 1995, were interviewed prospectively and followed up over a minimum of 18 months. Data were obtained from the Area Health Service's computerised booking system and patient self-report. RESULTS: Relevant surgical specialty, urgency rating, employment status and health insurance status were significantly associated with waiting time. Age, hospital, sex, country of birth, education, marital status and holding a Health Care Card were not significantly associated with waiting time. Self-reported health status (as measured by the SF-36) was not associated with waiting time for surgery. CONCLUSIONS: Waiting time for surgery was not simply determined by how urgently patients need surgery, but also by the type of surgery needed and patient's employment and health insurance status. The extent of disability experienced by patients, as measured by the SF-36, was not reflected in waiting times for surgery. PMID- 9847898 TI - HIV disease progression in Australia in the time of combination antiretroviral therapies. AB - OBJECTIVE: To examine the effect of recent developments in antiretroviral therapy on HIV disease progression and survival. DESIGN: Retrospective cohort study. PARTICIPANTS AND SETTING: Two cohorts of people with HIV were defined retrospectively from the records of a large immunology laboratory. The first cohort were subjects whose CD4+ T cell counts had dropped to 200 x 10(6)/L during 1990, and the second were subjects whose CD4+ T cell counts had dropped to 200 x 10(6)/L in 1994. MAIN OUTCOME MEASURES: HIV disease progression and survival was determined over a minimum three years of follow-up for each cohort (i.e., 1990 1993; 1994-1997). RESULTS: 346 subjects were included in the analysis (193 subjects from 1990 and 153 from 1994). The relative risk of progression to AIDS in the 1994 cohort compared with the 1990 cohort was 0.57 (95% confidence interval, 0.35-0.91; P = 0.018) and the relative risk of death was 0.20 (95% confidence interval, 0.08-0.49; P < 0.001). CONCLUSIONS: There were 43% fewer AIDS cases and 80% fewer deaths in the time following the increased availability of combination antiretroviral therapy in Australia. PMID- 9847899 TI - Exacerbation of the inflammatory response to Mycobacterium tuberculosis after antiretroviral therapy. AB - A patient with HIV infection was successfully treated for pulmonary tuberculosis, but pulmonary inflammation and lymphadenitis worsened dramatically after subsequent combination antiretroviral therapy. As this relapse coincided with development of a strong delayed-type hypersensitivity response to tuberculin and improved after treatment with the anti-inflammatory agent oxpentifylline, it was probably caused by restoration of pathogen-specific cellular immunity. PMID- 9847900 TI - Who cares for junior medical officers? AB - Recent changes in the way that some hospitals are run have created particular problems for junior medical staff. Here we describe a new system of managing such staff, under evaluation at a large Sydney teaching hospital, which seems to be working well and which has been positively appraised by the Postgraduate Medical Council and the Australian Council on Healthcare Standards. PMID- 9847901 TI - Cardiovascular ageing and heart failure. AB - Increase in arterial stiffness with age causes elevation of systolic blood pressure, which is the most common antecedent of heart failure in older people. Heart failure results from systolic and diastolic dysfunction; in either case, reducing mechanical load is the basis for preventing and treating heart failure. PMID- 9847902 TI - Eating disorders revisited. II: Bulimia nervosa and related syndromes. AB - Bulimia nervosa and related syndromes are common, and occur in up to 5% of women who attend general practitioners. Young women in First World countries, particularly those who "diet", are at increased risk. Behaviours, such as binge eating and induced vomiting, are typically kept well hidden. Only a minority of those with these disorders present for treatment. General practitioners play a key role in primary and secondary prevention. Effective treatments include psychotherapies that focus on the patient's attitudes and relationships, not just the binge eating behaviour. About 50% of patients make a complete recovery, but the long term outcome is unknown. PMID- 9847903 TI - Colorectal cancer. AB - Colorectal cancer is the leading cause, after lung cancer, of death from cancer in Australia. About 50% of colorectal cancers are attributed to dietary factors and about 15%-20% to genetic factors, including high risk familial syndromes. The most common presenting symptoms are rectal bleeding (left-sided cancers) and iron deficiency anaemia (right-sided cancers). Patients with suggestive symptoms should have a digital rectal examination, followed by sigmoidoscopy and biopsy if a rectal lesion is suspected, or colonoscopy if colon cancer is suspected. Treatment of most colorectal cancers is segmental resection with a wide resection margin; the role of adjuvant therapy is still being refined. Screening with annual (or biennial) faecal occult blood tests is recommended for people over 50 years without symptoms and with average or slightly above average risk; more intensive, earlier screening is recommended for those with greater risk. PMID- 9847904 TI - Suicide among young Australians, 1964-1993: an interstate comparison of metropolitan and rural trends. PMID- 9847905 TI - Suicide among young Australians, 1964-1993: an interstate comparison of metropolitan and rural trends. PMID- 9847906 TI - Eosinophilic leukaemoid reaction and interleukin-5 in metastatic melanoma. PMID- 9847907 TI - Dissecting the rugby tackle: do shoulder pads contribute to cervical spine injuries? PMID- 9847908 TI - Australian medical research: more resources and the right balance. PMID- 9847910 TI - Alcohol licensing data. PMID- 9847909 TI - Massive gastric dilatation after a bulimic excess. PMID- 9847911 TI - Viagra--beware the quick fix. PMID- 9847912 TI - Putting patient care first when group practices break down. PMID- 9847913 TI - The measles tragedy revisited. PMID- 9847915 TI - Science-based, unified approach needed to safeguard the nation's food supply. PMID- 9847914 TI - Neuroscience and public policy. PMID- 9847916 TI - AHCPR picks two clinical prevention centers. PMID- 9847917 TI - Advisory council calls for development of new vaccines for tuberculosis. PMID- 9847918 TI - DHHS, EPA Fund. Children's environmental health research centers. PMID- 9847919 TI - No scientific justification to suspend hepatitis B immunization. PMID- 9847920 TI - A year of living dangerously: the tobacco control community meets the global settlement. AB - Momentum toward Congressional action on tobacco issues began with the announcement in June 1997 of a proposed "global tobacco settlement" between the tobacco industry, a group of state Attorneys General, and private class action lawyers. For the next year, tobacco issues received unprecedented national attention, culminating in the U.S. Senate's consideration and ultimate defeat of the McCain tobacco bill. Through the proposed settlement, the Attorneys General and others involved in talks with the industry sought to reduce tobacco use by attempting to forge a "win-win" solution for all parties. In exchange for money and public health concessions, the industry would be granted sweeping protection from litigation and thus a stable environment in which to operate. Members of the public health community responded to the "global tobacco settlement" in very different ways. The authors explore how different visions of possibilities and practicalities were played out in the fight for strong Federal tobacco control legislation and attempt to draw lessons for the future. PMID- 9847921 TI - Constructing violence as a public health problem. AB - Once viewed primarily as a criminal justice problem, violence and its prevention are now often claimed by public health professionals as being within their purview. The author reviewed 282 articles published in public health and medical journals from 1985 through 1995 that discussed violence as a public health problem. She found that while authors tended to identify social and structural causes for violence, they suggested interventions that targeted individuals' attitudes or behaviors and improved public health practice. Her study illuminates the tension between public health professionals' vision of the social precursors of violence and their attempts to apply a traditional set of remedies. In targeting individuals to rid the nation of violence, the public health community is deemphasizing societal causes. PMID- 9847922 TI - The selling of olestra. AB - The Procter & Gamble Company spent 30 years and an estimated $500 million to bring its non-digestible fat substitute, olestra, to market. The Food and Drug Administration approved olestra as a food additive but requires products containing olestra to carry a warning statement about its potential effects on gastrointestinal function. In obtaining approval for olestra, P&G conducted a lengthy, persistent, and comprehensive campaign to enlist support from members of Congress; FDA staff; and food, nutrition, and health professionals. This campaign raises larger questions about corporate influence on government policies, and the relationships of corporations to health professionals. To address these larger concerns, the author reviews the history of olestra's approval; describes P&G's campaign to obtain support from FDA and Congress, to defend olestra against critics, and to market it to professionals, the press, and consumers; and suggests implications for public health policies. PMID- 9847923 TI - Estimating vaccination coverage using parental recall, vaccination cards, and medical records. AB - OBJECTIVE: To compare estimates based on vaccination cards, parental recall, and medical records of the percentages of children up-to-date on vaccinations for diphtheria, tetanus, and pertussis; polio; and measles, mumps, and rubella. METHOD: The authors analyzed parent interview and medical records data from the Baltimore Immunization Study for 525 2-year-olds born from August 1988 through March 1989 to mothers living in low-income Census tracts of the city of Baltimore. RESULTS: Only one-third of children had vaccination cards; based on medical records, these children had higher up-to-date coverage at 24 months of age than did children without cards. For individual vaccines, only two-thirds of parents could provide information to calculate coverage rates; however, almost all provided enough information to estimate coverage for the primary series. For each vaccine and the series, parental recall estimates were at least 17 percentage points higher than estimates from medical records. For children without vaccination cards whose parents could not provide coverage information, up-to-date rates based on medical records were consistently lower than for children with cards or with parents who provided coverage information. CONCLUSIONS: Population-based vaccine coverage surveys that rely on vaccination cards or parental recall or both may overestimate vaccination coverage. PMID- 9847925 TI - Optimal disease prevention using vaccination depends on a system. PMID- 9847924 TI - Deficiencies in current childhood immunization indicators. AB - OBJECTIVE: To investigate "up-to-date" and "age-appropriate" indicators of preschool vaccination status and their implications for vaccination policy. METHODS: The authors analyzed medical records data from the Baltimore Immunization Study for 525 2-year-olds born from August 1988 through March 1989 to mothers living in low-income Census tracts of the city of Baltimore. RESULTS: While only 54% of 24-month-old children were up-to-date for the primary series, indicators of up-to-date coverage were consistently higher, by 37 or more percentage points, than corresponding age-appropriate indicators. Almost 80% of children who failed to receive the first dose of DTP or OPV age-appropriately failed to be up-to-date by 24 months of age for the primary series. CONCLUSIONS: Age-appropriate immunization indicators more accurately reflect adequacy of protection for preschoolers than up-to-date indicators at both the individual and population levels. Age-appropriate receipt of the first dose of DTP should be monitored to identify children likely to be underimmunized. Age-appropriate indicators should also be incorporated as vaccination coverage estimators in population-based surveys and as quality of care indicators for managed care organizations. These changes would require accurate dates for each vaccination and support the need to develop population-based registries. PMID- 9847926 TI - Silicone gel breast implant adverse event reports to the Food and Drug Administration, 1984-1995. AB - OBJECTIVES: To characterize the adverse event reports on silicone gel breast implants (SGBIs), including death reports, submitted to the Food and Drug Administration (FDA) from 1984 through 1995 and to analyze changes in the type and complexity of reports following extensive media coverage of breast implants. METHODS: The authors analyzed mandatory and voluntary reports from the adverse events reporting system for medical devices at the FDA. RESULTS: In 1988, adverse event reports related to SGBIs accounted for 2.4% of the 14,473 mandatory reports entered into the FDA database on medical devices. In 1992, SGBI-related reports accounted for 30.3% of the total 66,476 mandatory reports of adverse events. The most frequently reported adverse event in 1988, before the widespread publicity on breast implants, was implant burst or rupture. In contrast, in 1992 the most frequently reported event was reaction, a term used to describe a range of adverse effects. CONCLUSIONS: The numbers of mandatory and voluntary reports of SGBI-related adverse events increased exponentially, as did the complexity of the reports, following publicity over the lack of safety data on breast implants and a short voluntary moratorium on their sale. A significant proportion of reports lacked information on specific medical symptoms or diagnoses. PMID- 9847927 TI - Hygienic practices and acute respiratory illness in family and group day care homes. AB - OBJECTIVE: To describe hygiene practices in licensed group day care and family day care homes and the association between these practices and the prevalence of respiratory illnesses in the children in attendance. METHODS: Self-administered surveys were mailed to 137 group and 204 family day care providers. RESULTS: Wearing diapers and being younger than age three were associated with a higher frequency of respiratory illness. Children attending family day care homes had more respiratory illness than children attending group day care homes. Infrequent washing of children's or providers' hands after nose wiping, after diapering, before meals, and before food preparation was significantly associated with a higher frequency of respiratory illness. Use of shared cloth towels instead of individual paper towels and washing of sleeping mats less than once a week were also associated with a higher frequency of respiratory illness. CONCLUSIONS: The findings underscore the importance of handwashing and other hygiene practices in reducing the spread of disease in day care settings. PMID- 9847928 TI - Tuberculosis among foreign-born residents of southern Florida, 1995. AB - OBJECTIVE: To examine the characteristics of foreign-born people with tuberculosis (TB) in Southern Florida, their contribution to the total number of TB cases, and available data on their HIV status as well as to determine the number of cases detected by the overseas medical screening of immigrants and refugees. METHODS: The authors reviewed TB cases reported by Broward, Dade, and Palm Beach counties in 1995. Case records were matched against the CDC Division of Quarantine database of immigrants and refugees suspected to have TB at the time of visa application overseas. RESULTS: Nearly half (49%) of TB cases in the three counties were among people born outside the United States--34% in Broward County, 58% in Dade County, and 40% in Palm Beach County. A high percentage (26%) were co-infected with HIV. Of those with known date of arrival, 68% had been in the United States for five or more years. Only three cases had been identified by overseas immigrant screening. CONCLUSIONS: A low percentage of TB cases in foreign-born people were identified through the overseas screening system. Controlling TB in South Florida will require efforts targeted toward high risk populations, including people with HIV infection. PMID- 9847929 TI - Sweden: preventing health workers' back injuries. PMID- 9847931 TI - Seoul Congress Abstracts. PMID- 9847930 TI - [52nd Congress of the German Society for Gynecology and Obstetrics. Nurnberg, 8 12 September 1998. Abstracts]. PMID- 9847932 TI - Childhood Obesity. Satellite symposium of the 8th International Congress of Obesity. Paris, France, 4 September 1998. Abstracts. PMID- 9847933 TI - British Human Genetics Conference 1998. September 28-30. Abstracts. PMID- 9847934 TI - 5th Central European Lung Cancer Conference. Prague, September 13-16, 1998. Abstracts. PMID- 9847935 TI - 21st Annual meeting of the Japan Neuroscience Society and 1st joint-meeting of the Japan Neuroscience Society and the Japanese Society for Neurochemistry. Tokyo, Japan, September 21-23, 1998. Abstracts. PMID- 9847936 TI - 8th Congress of the European Shock Society. La Hulpe, Belgium, October 15-17, 1998. Abstracts. PMID- 9847937 TI - The changing challenges in the lives of three generations of professional women. PMID- 9847938 TI - Postmodernizing the unconscious with the help of Derrida and Lyotard. PMID- 9847939 TI - The evolution of mutuality in the resolution of transference. PMID- 9847940 TI - Voicing voicelessness: on the poetics of faith. PMID- 9847941 TI - A critique of classical psychoanalytic theories of anti-Semitism: a commentary on M. Ostrow's myth and madness: the psychodynamics of anti-Semitism. PMID- 9847942 TI - Timeliness of laboratory investigations in detecting and managing infectious diseases. PMID- 9847943 TI - Helicobacter pylori and peptic ulcer disease: has the emperor got no clothes? PMID- 9847944 TI - Glycoprotein IIb/IIIa inhibitors, surgery and bleeding. PMID- 9847945 TI - Seroprevalence of Helicobacter pylori in a longitudinal study of New Zealanders at ages 11 and 21. AB - BACKGROUND: Helicobacter pylori seroprevalence increases with age in adult life but spontaneous reversion may occur in childhood and adolescence. AIMS: To determine the seroprevalence of H. pylori in a longitudinal study of New Zealanders at ages 11 and 21. METHODS: Serum from members of the Dunedin Multidisciplinary Health and Development Study (DMHDS) at age 11 (n = 561; 303 males, 258 females) and 21 (n = 785; 413 males, 372 females) was tested for H. pylori antibodies. A large proportion of those tested at age 11 was retested at age 21 (n = 465; 262 males, 203 females). Serological status was examined in terms of gender, socioeconomic status (SES) and self-reported use of antibiotics. RESULTS: The seroprevalence of H. pylori decreased by 38% from 6.6% (37/561) at age 11 to 4.1% (32/785) at age 21. Seroprevalence at age 11 was not associated with gender or SES. For those tested at both ages, the drop in seroprevalence from 6.7% to 4.1% was statistically significant (t = 2.57, p < 0.01, paired t test) and was much greater in females (71%) than males (12%). Of the 31 seropositive individuals at age 11, 17 (six males, 11 females) seroreverted and self-reported antibiotic use in the year preceeding age 21 was more common in females (eight/11) than males (zero/six). Of the 434 seronegative individuals at age 11, only five (four males, one female) had seroconverted at age 21. CONCLUSIONS: Seroprevalence in the DMHDS declined from age 11 to 21 predominantly in females. The decline involved a greater rate of seroreversion and lower rate of seroconversion in females than males. PMID- 9847946 TI - Australian encephalitis in the Northern Territory: clinical and epidemiological features, 1987-1996. AB - BACKGROUND: The last epidemic of Australian encephalitis occurred in 1974. Since then, cases have been reported from the Kimberley of Western Australia (WA). AIMS: To describe the epidemiology and clinical features of Australian encephalitis in the Northern Territory (NT) of Australia. METHODS: Review of cases of Australian encephalitis presenting to Royal Darwin Hospital from 1987 1996 and review of sentinel chicken surveillance for Australian encephalitis viruses. RESULTS: Sixteen patients were identified; ten from the NT and six from WA. Cases occurred in the years 1987, 1988, 1991 and 1993. Infection was acquired throughout northern NT below latitude 20 degrees S in the months March to July. All infections were due to Murray Valley encephalitis (MVE) virus. Eleven of the patients were children. Distinguishing features were spinal cord and brainstem involvement and the absence of seizures in adults. CT scanning was normal and EEG showed no focal activity. Five died (31%) and four (25%) have residual neurological disability. Sentinel chicken surveillance since 1992 shows yearly seroconversion to MVE virus throughout northern NT; human cases occurred simultaneously with chicken seroconversion in 1993. CONCLUSIONS: Australian encephalitis is endemic in the NT; the areas at risk are north of Tennant Creek. Outbreaks are seasonal and occur every few years. Young children are most at risk. Mortality and morbidity are high. Prevention of disease is by avoidance of mosquito exposure and vector control measures. PMID- 9847947 TI - Evaluation of radiation dose and risk to the patient from coronary angiography. AB - BACKGROUND: Coronary angiography is a commonly performed diagnostic procedure with life saving benefits for the patient. However, because of its potential to deliver a high radiation dose, caution is warranted. AIM: The purpose of this study was to determine the effective dose to an average patient from coronary angiography, to estimate the associated radiation risk in terms of fatal malignancy, and to examine ways effective dose may be reduced. METHOD: The dose area product data from 210 examinations was used to determine the effective dose to the average patient by means of a commercially available software programme. Subsequently, the effective dose was used to estimate radiation risk based on the population averaged probability coefficients given in ICRP 60 and contrasted against age-related risk coefficients from the NRPB. The components of effective dose and radiation risk were examined to identify the organs most at risk. RESULTS: The average effective dose was found to be 3.4 +/- 1.3 mSv. The risk of fatal malignancy was estimated as 170 per million cases, or around one in 6000. The average lung dose was 14.7 mSv, with the risk of fatal lung cancer estimated as 125 per million cases or one in 8000. CONCLUSIONS: The average effective dose in this study was less than half that of most other studies. The lung was identified as the critical organ and the estimated risk of fatal cancer amounted to an increase of 0.017% above the normal incidence for the general population. The study affirmed that using a reduced cine frame rate, low dose pulsed fluoroscopy, added copper filtration and a conscious effort to reduce screening time can significantly reduce patient dose. PMID- 9847948 TI - Diabetic ketoacidosis in adults at Auckland Hospital, 1988-1996. AB - BACKGROUND: Diabetic ketoacidosis (DKA) is associated with significant morbidity and mortality. Recent evidence suggests that patients with both type 1 and type 2 diabetes can develop DKA. AIM: To review the experience in managing patients admitted to Auckland Hospital with DKA over an eight year period. METHODS: A retrospective chart review was undertaken to identify patients with a discharge code of DKA admitted to Auckland Hospital between May 1988 and October 1996. RESULTS: One hundred and twenty-five patients were identified who met the defined criteria for DKA. The in-patient mortality for the group was 2.4%. Thirteen patients (10.4%) probably had type 2 diabetes. Thirty-eight (30.4%) patients were admitted to the Department of Critical Care Medicine (DCCM)--these patients had a significantly lower systolic blood pressure and arterial pH, together with a significantly higher admission blood glucose and longer duration of insulin infusion than those not admitted to DCCM. Following their index admission 25% of patients were readmitted to hospital with DKA during the study period. Errors in insulin self-administration that contributed to admission to hospital with DKA were identified in 61% of the patients with known diabetes. CONCLUSIONS: Patients with DKA in this study spent about a week in hospital and a significant proportion were admitted to the DCCM. In spite of this the overall mortality was low. Many of these patients were readmitted to hospital with DKA. A small number of patients with DKA may have type 2 diabetes and may not need long term insulin therapy. More effort on patient education regarding insulin use with illness, may prevent admission to hospital with DKA. PMID- 9847949 TI - Increased survival in patients diagnosed with Hodgkin's disease in Tasmania, 1972 1992. AB - BACKGROUND: It has been shown that in certain populations the prognosis of Hodgkin's disease (HD) has improved markedly since the late 1960s. This has not been formally demonstrated in an Australian population. AIMS: To review all patients in Tasmania diagnosed with HD between 1972 and 1992, and to ascertain whether variation in survival is evident in this group over this period. METHODS: Tasmanian patients with HD diagnosed from 1978 to 1992 were identified retrospectively from the Tasmanian Cancer Registry database. Identification of those diagnosed prior to 1978 was obtained from a previously published data set. To be valid for inclusion, subjects were required to have been diagnosed between January 1972 and December 1992, enabling a minimum four year follow up period. Survival was assessed by contacting patients' medical practitioners and by examining the most current electoral roll, medical records, and the register of births, deaths and marriages. Univariate and multivariate analyses were performed of the influence on prognosis of age, sex, histological subtype and epoch of diagnosis; information concerning stage of disease was not available. RESULTS: During the period of this study 206 patients were newly diagnosed as having HD. Comparisons of cases diagnosed in the successive seven-year epochs 1972-8, 1979 85 and 1986-92 revealed a significant increase in survival duration (p = 0.023), with ten year survival rates of 46%, 55% and 73% respectively. In a multivariate analysis adjusting for age, sex and histology, each successive epoch was associated with an estimated 28% reduction in the death rate relative to the preceding epoch (p = 0.022). CONCLUSIONS: There was a significant improvement in the survival duration of patients diagnosed with HD in Tasmania over the period 1972-92, which was possibly due to a combination of better diagnostic techniques and more effective treatments. PMID- 9847950 TI - The effect of test duration on the sensitivity and specificity of ultra-rapid urease test for the detection of Helicobacter pylori infection. AB - BACKGROUND: 'Home made' ultra-rapid urease tests are used extensively in the Asia Pacific region. Data on the reliability of these 'home made' tests are limited. AIMS: To evaluate the effect of test duration on the sensitivity and specificity of a self-prepared biopsy urease test for the detection of Helicobacter pylori. METHODS: Using histology as the 'gold standard', the effect of test duration on the sensitivity and specificity of a self prepared urease test for the detection of H. pylori was evaluated in 411 consecutive patients undergoing upper gastrointestinal endoscopy. RESULTS: Histology was positive for H. pylori in 217 of the 411 patients (52.8%). Within 24 hours of retrieving the specimen, 189 (87.1%) of the histology positives and 174 (89.7%) of the histology negatives were correctly identified by the urease test. Of the H. pylori positives, 72.0%, 81.0%, 89.9% and 100% were detected by the urease test within one minute, five minutes, three hours and 24 hours respectively. Thus, sensitivities of the urease test at one minute, five minutes, three hours and 24 hours were 62.7%, 70.5%, 78.3% and 87.1% respectively. Corresponding figures for the specificity were 93.8%, 93.3%, 92.3% and 89.7% respectively. Using a receiver-operating characteristic curve, an optimal combination of sensitivity and specificity was obtained when the urease test was read at 24 hours. CONCLUSIONS: While the biopsy urease test was positive in most cases within a minute, better results could be obtained if the test continued to be read over a 24 hour period. PMID- 9847951 TI - Transjugular intrahepatic portosystemic shunt (TIPS) for the treatment of refractory ascites. AB - BACKGROUND: The Transjugular Intrahepatic Portosystemic Shunt (TIPS) corrects portal hypertension and has proven to be effective in controlling variceal bleeding in patients with cirrhosis. Several reports have now appeared suggesting a possible role in patients with refractory ascites. AIMS: To examine the outcome of TIPS for the treatment of refractory ascites in patients with cirrhosis. METHODS: Fifteen patients underwent TIPS for ascites between April 1992 and December 1996. The clinical findings, response to treatment, complications, shunt patency and survival of these patients were analysed. RESULTS: TIPS was successfully placed in all patients. The mean period of follow-up was 375 days (range: 14-1165 days). In eight patients (53%) there was a reduction in the degree of ascites after shunt insertion, with six patients (40%) having complete resolution. Age, Child-Pugh class or portal pressure gradient, before or after the procedure, were not predictive of response. Of five patients with renal insufficiency (serum creatinine > 130 umol/L), only one had improvement in ascites control. Six patients (40%) required shunt revision during follow-up, either for acute thrombotic occlusion (two) or stent stenosis (four). New or worsening encephalopathy developed in ten patients (67%). Two patients (13%) died of liver failure within 30 days. Cumulative survival was 46% at one year and 18% at two years. Treatment response was associated with increased survival (p = 0.02), with median survival of 658 days as compared with 71 days for treatment failure. CONCLUSIONS: TIPS can be effective in the treatment of refractory ascites in patients with cirrhosis. Our experience suggests the benefit may be less for patients with advanced liver disease and renal impairment. Controlled trials are needed to compare TIPS with other treatment modalities such as large volume paracentesis or peritoneovenous shunting. PMID- 9847952 TI - Prevalence and vascular associations with migraine in older Australians. AB - BACKGROUND: Migraine is a common disorder with recently described vascular associations, yet there are few Australian population-based data describing migraine prevalence. AIMS: To assess the prevalence and vascular associations with lifetime past history of typical migraine headache in a representative sample of older Australians. METHODS: The Blue Mountains Eye Study examined 3654 permanent residents aged 49 or older living in two postcode areas, west of Sydney (82.4% participation) during 1992-4. A structured interview was administered, including questions about past or present history of typical migraine. The diagnosis was consistent with International Headache Society criteria. RESULTS: A lifetime past history of typical migraine was given by 17% of participants, including 22% of women and 10% of men, a female:male ratio of 2.3:1. A marked trend for declining lifetime migraine frequency with increasing age was found for both sexes. Modest statistically significant associations were found with vascular disease history, after multivariate adjustment, which included vascular risk factors. These associations were stronger in men than in women. Among men, typical migraine was significantly associated with history of angina, odds ratio (OR) 2.0, acute myocardial infarction (OR 1.9) and stroke (OR 2.2). Among women, statistically significant associations were present only with history of myocardial infarct (OR 1.8). CONCLUSIONS: These data indicate similar prevalence rates for lifetime typical migraine history in a representative sample of older Australians, compared to recent US and Canadian populations. Modest, statistically significant associations between typical migraine and past history of vascular disease were found, with the strongest associations found in men. PMID- 9847953 TI - Prevalence and demographic determinants of metronidazole resistance by Helicobacter pylori in a large cosmopolitan cohort of Australian dyspeptic patients. AB - BACKGROUND: The pre-treatment sensitivity of Helicobacter pylori to metronidazole is a key determinant of successful eradication therapy and should influence local choice of therapy. However, there are few data defining the prevalence of metronidazole resistance (MR) in Australia. AIM: To determine prospectively the prevalence and demographic determinants of MR in H. pylori isolates from a large and cosmopolitan cohort of dyspeptic patients in Sydney. METHODS: Consecutive dyspeptic patients undergoing endoscopy had gastric biopsies for histology, urease test and culture. Metronidazole resistance was determined by E-test after subculture. An MIC > 8 micrograms/mL defined MR. Patient age, gender, birthplace and history of previous nitroimidazole use were recorded. RESULTS: In 732 patients, H. pylori was present in 46.4%. Culture was successful in 81% and subculture for MR in 88% of these. In 237 evaluable patients the overall MR rate was 59.1%. Five patients had had prior triple therapy for H. pylori (of which four of five had MR). Therefore, the primary MR rate in the study population was 58.6% (136/232). MR was more prevalent in younger patients (p = 0.0002). The MR rate was 70.4% in patients 18-39 years, 66.7% in those aged 40-59 years and lowest (38.9%) in those 60 years or older (p = 0.002). The MR rate was highest in patients born in Southeast Asia (72.8%, 59/81) and significantly higher than in Australian born (48.1%, 26/54), or Southern European (46.2%, 24/52) born patients (p = 0.002). There was no gender difference. Logistic regression to determine the impact of each variable (birthplace, age and gender) on MR identified Southeast Asia birthplace as a factor associated with greater likelihood of harbouring an MR isolate (OR 1.88, p = 0.02). Southern European born patients had the lowest risk of MR (OR 0.70, p = 0.02) as did patients older than 60 years (OR 0.56, p = 0.04). A definite history of prior metronidazole use was infrequent and not predictive of MR. CONCLUSIONS: While a high rate of MR is not unexpected in patients born in developing countries, the high rate in Australian born patients is surprising and of concern. This may relate to the high local usage of nitroimidazoles as monotherapy and has important implications for the effectiveness of metronidazole containing triple therapies. PMID- 9847954 TI - Sunscreens: are Australians getting the good oil? PMID- 9847955 TI - Thyroid disease and pregnancy. PMID- 9847956 TI - The use of percutaneous transluminal angioplasty (PTA) for the treatment of extracranial atherosclerotic vascular disease. The Australian Association of Neurologists (AAN). PMID- 9847957 TI - Hydroxyurea-induced fever and hepatitis. PMID- 9847958 TI - Serotyping delays and implications for public health action: The Northern Territory experience of the 1996 national outbreak of Salmonella Mbandaka and a comparison with Western Australia. PMID- 9847959 TI - Helicobacter pylori infection and duodenal ulceration. PMID- 9847960 TI - Vertebral osteomyelitis and discitis associated with Abiotrophia adiacens (nutritionally variant streptococcus) infection. PMID- 9847961 TI - Mexiletine as a treatment for malignant neuropathic pain. PMID- 9847962 TI - Sleep apnoea in hypothyroidism. PMID- 9847963 TI - Chylothorax from radiation-induced mediastinal fibrosis. PMID- 9847964 TI - Thrombophilia in a young Down syndrome patient. PMID- 9847965 TI - Pulmonary eosinophilia associated with aminoglutethimide. PMID- 9847966 TI - Cephalic tetanus. PMID- 9847967 TI - The forgotten granuloma. PMID- 9847968 TI - Striking a balance in drug information--the Australian Medicines Handbook. PMID- 9847969 TI - More pharmaceutical company influence? PMID- 9847970 TI - Nondiabetic peripheral neuropathies. AB - The evaluation of a peripheral neuropathy can be challenging. The history remains the most informative portion of the evaluation. Electrodiagnostic testing is important to characterize the underlying pathology. Laboratory tests should be ordered based on clinical findings. Referral to neuromuscular specialists may be helpful if the diagnosis is not forthcoming, or if ongoing treatment is required. A sizable proportion of neuropathies cannot be diagnosed. For these cases, uncomfortable symptoms can be managed successfully. PMID- 9847971 TI - An approach to clinical acid-base problem solving. AB - This article demonstrated a systematic approach to acid-base problem solving through the application of the so-called "Rules of Five." This technique was compared with other options for acid-base problem solving, namely, clinicians' use of gestalt or an acid-base map. The superiority of the systematic approach in uncovering triple acid-base disorders was shown. PMID- 9847972 TI - The treatment of Parkinson's disease: current concepts and rationale. AB - The recent advances in the treatment of Parkinson's disease have made for significant improvements in the quality of life and mortality rate of those who suffer from this neurodegenerative disease. At the same time, the number of options and the complexity of multi-drug regimens have posed a great challenge for the clinician caring for the patient with Parkinson's disease. Though there are still many questions to be answered in regard to the potential neuroprotective effects of several medications, a few general rational treatment plans can be outlined. In patients requiring treatment in the early stages of the disease, especially with a predominance of tremor, anticholinergics or amantadine should be considered initially. At this point, it would be reasonable to add selegiline for both therapeutic and possible neuroprotective effects. As a patient becomes more affected by the disease and additional therapy is necessary, starting either a dopamine agonist or levodopa would be a rational choice. Continuation of selegiline and, possibly, amantadine for neuroprotective reasons should be contemplated. Titration in levodopa therapy (with controlled-release or standard levodopa) to higher levels should prompt addition of a dopamine receptor agonist if one has not been started previously. Conversely, if a patient is receiving only a dopamine receptor agonist and is becoming progressively disabled, levodopa should be added to the regimen. Fluctuations in motor abilities may be improved further by the use of a COMT inhibitor. Patients with uncontrollable motor fluctuations should be considered for surgery. Undoubtedly, the coming years will bring more treatment options and more evidence on which sequences and combinations of therapies are the most beneficial. Differences in efficacy and adverse effects for each patient must be taken into consideration when outlining and carrying out a treatment plan. By using a rational approach to the treatment of Parkinson's disease, with the above guidelines in mind, the patient should be able to enjoy a good quality of life and level of function for many years. PMID- 9847973 TI - Interpretation of thyroid function tests: an update. PMID- 9847974 TI - Advances in the diagnosis and management of scleroderma-related vascular complications. AB - Vascular disease is observed in virtually all patients with systemic sclerosis and presents several challenges for the clinician. The alterations in vascular tone noted with Raynaud's phenomenon may have corollaries in both the pathogenesis and management of systemic sclerosis-related internal organ complications. Early recognition of isolated pulmonary hypertension, coronary microcirculatory disease, and scleroderma renal involvement may be the keys to successful outcomes. Future trends in the management of systemic sclerosis may be directed at preventing the development of these vascular syndromes. This may occur through combined therapies directed at the abnormal immune response, disregulated fibroblast function, and resultant abnormal vascular response. PMID- 9847975 TI - Adenocarcinoma of the stomach: an update. PMID- 9847976 TI - Diagnosis and monitoring of spondyloarthropathy. PMID- 9847977 TI - Escalating health care costs: managed care organizations. PMID- 9847978 TI - Abdominal obesity and its metabolic complications: implications for the risk of ischaemic heart disease. AB - Although the health hazards of obesity are well established, obese individuals are not all at equal risk of developing a disease, which reflects the heterogeneity of this condition. The regional distribution of body fat is now recognized as a very important component of the obesity-related health hazards. Epidemiological studies have shown that abdominal obesity, that is, a preponderance of fat in the abdominal area, is a better predictor of both cardiovascular disease and type 2 diabetes than obesity per se. It is now generally accepted that the fat located within the abdominal cavity, the visceral fat, is the best correlate of most of the highly atherogenic metabolic complications seen in individuals with abdominal obesity. These include, among others, insulin resistance and hyperinsulinaemia, hypertriglyceridaemia, reduced plasma high-density lipoprotein (HDL) cholesterol concentrations and an increased number of small, dense low-density lipoprotein (LDL) particles. This review summarizes the evidence that these metabolic complications may account to a large extent for the increased risk of cardiovascular disease associated with abdominal/visceral obesity. Abdominal obesity may be the most prevalent denominator of highly atherogenic dyslipidaemic and hyperinsulinaemic/insulin resistant states in affluent, sedentary societies. Targeting individuals with this high-risk trait in primary prevention is therefore crucial if we are truly to have an impact on the incidence of cardiovascular disease. PMID- 9847979 TI - Abdominal obesity and cardiovascular risk in children. PMID- 9847980 TI - Abdominal obesity and associated cardiovascular comorbidities in the elderly. PMID- 9847981 TI - Body weight loss and maintenance with physical activity and diet. PMID- 9847982 TI - Menopause, central body fatness, and insulin resistance: effects of hormone replacement therapy. AB - In addition to being associated with termination of reproductive life in women, the menopause coincides with an increase in several comorbidities including cardiovascular disease. This increase in the prevalence of cardiovascular disease in the postmenopausal years has been partially attributed to adverse effects of estrogen deficiency on plasma lipid-lipoprotein levels and on the cardiovascular system, although other factors are contributing. Central body fatness and insulin resistance are components of a cluster of metabolic abnormalities which also increases the risk of cardiovascular disease. This review summarizes studies that have examined the effects of the menopause transition and of estrogen-replacement therapy on central body fatness and insulin resistance. Review of cross-sectional studies suggests that the menopause transition is associated with an increase in abdominal and visceral adipose tissue accumulation, as measured either with dual X-ray absorptiometry or computed tomography. These results appear to be independent of the aging process and total body fatness. In general, cross sectional studies using circumference measurements did not find any significant effect of the menopause. Longitudinal studies also support that accumulation of central body fatness accelerates with menopause. The effects of the menopause on insulin resistance appear to be moderate, if any, although available studies are clearly insufficient to draw firm conclusions. The majority of interventional studies support the notion that hormone-replacement therapy attenuates the accumulation of central fat in postmenopausal women, compared with control or placebo-treated women. Retrospective comparisons of hormone users and nonusers also support a protective effect of hormone replacement on fat distribution. Moderate effects of estrogen therapy were found on insulin resistance in postmenopausal women, although long-term, controlled trials using accurate measurements of insulin sensitivity are lacking. Treatment with progestins exerts moderate deleterious effects on insulin sensitivity, which may be attributable to the partial androgenicity of progestins used. It is concluded that part of the increased incidence of cardiovascular disease in postmenopausal women may be attributable to increased central body fatness. Therapies aiming at preventing these changes in fat distribution such as hormone-replacement therapy, diet or exercise are likely to provide long-term cardiovascular and metabolic benefits for women's health. PMID- 9847983 TI - Serum concentration of calcium, 1,25 vitamin D and parathyroid hormone are not correlated with coronary calcifications. An electron beam computed tomography study. AB - BACKGROUND: The amount of coronary artery calcification, measured using electron beam computed tomography, is correlated with the volume of coronary artery atherosclerotic plaque, the severity of stenosis by angiography, and with the likelihood of future cardiovascular events. The deposition of calcium in atherosclerotic plaques may also be influenced by determinants of calcium metabolism, thus contributing to the variance of the relation between coronary artery calcification and extent of atherosclerosis. Our objective was to determine whether this variance could be explained by differences in the parameters of calcium metabolism. DESIGN AND METHODS: We measured serum concentrations of calcium, 1,25(OH)2 vitamin D and parathyroid hormone (PTH) in 50 subjects undergoing angiography for clinical indications, and evaluated the correlations between these concentrations and calcium deposition in the coronary arteries, and the ratio of calcium deposition to extent of atherosclerosis using coronary angiography. RESULTS AND CONCLUSIONS: Serum concentrations of calcium 1,25(OH)2 vitamin D and PTH were not correlated with coronary calcification or the ratio of coronary calcification to the extent of coronary stenosis. We conclude that, in subjects undergoing coronary angiography, the variance of the relationship between coronary atherosclerosis and coronary calcium is not a result of differences in serum concentrations of calcium, 1,25(OH)2 vitamin D or PTH. PMID- 9847985 TI - Paradigm shift in mechanical ventilatory support for acute respiratory distress syndrome: do no harm. PMID- 9847984 TI - Improvement in quality of life during 5 years after coronary artery bypass grafting. AB - OBJECTIVE: To describe the changes in various aspects of quality of life (QOL) from before coronary artery bypass grafting (CABG) to 5 years after the procedure. PATIENTS AND METHODS: Patients who underwent CABG in the western region of Sweden in 1988-1991 were approached with questionnaires evaluating their QOL prior to and 3 months and 1, 2, and 5 years after the operation. Three different instruments were used: the Nottingham Health Profile, the Psychological General Well-Being Index, and the Physical Activity Score. RESULTS: In all 2121 patients underwent CABG, of whom 310 died during 5 years of follow-up. With all three instruments QOL had improved 5 years after CABG compared with prior to the operation. However, all three instruments revealed a slight but significant deterioration in estimated QOL between 2 and 5 years after CABG. CONCLUSIONS: QOL 5 years after CABG is better than that prior to the operation, but between 2 and 5 years after the operation a slight deterioration in QOL is observed. PMID- 9847987 TI - Bibliography current world literature. PMID- 9847988 TI - Solid landmarks amidst chaos. PMID- 9847986 TI - Feasibility and safety of percutaneous laser revascularization using the Biosense system in porcine hearts. AB - BACKGROUND: Direct myocardial revascularization (DMR) is being explored to improve symptoms in patients with refractory ischemic coronary syndromes. This study assessed the safety and feasibility of percutaneous DMR using the Biosense non-fluoroscopic navigation system (incorporating the holmium : yttrium aluminium garnet laser) in pig hearts. METHODS AND RESULTS: Twenty pigs underwent left ventricular mapping using the Biosense system, followed by percutaneous DMR with a holmium : yttrium aluminium garnet laser at 2 J x 1 pulse, and were sacrificed acutely (n = 5), at 24 h (n = 5), and at 3 weeks (n = 10) after the DMR procedure. The hearts were examined grossly and microscopically. Serial creatine kinase-MB level was measured up to 24 h. There were no procedural complications. Animals received 13 +/- 2 channels in 19 +/- 7 min. The catheter was able to reach all regions, including the septum. There was no increase in creatine kinase MB level up to 24 h. One animal died within an hour of the procedure. There was one sealed perforation into the subepicardial fat as a result of same site laser activation. Histologic evaluation in the acute and 24 h groups revealed laser channels which were 3.6 +/- 2.2 mm long, 1.5 +/- 0.7 mm wide, with an entry angle of 73 +/- 12 degrees. Channels were filled with platelet thrombi acutely and were surrounded by well-defined rims of thermal coagulation and an 'impact zone' of viable myocardium; at 3 weeks, no channels remained patent and they had been replaced by well-healed granulation tissue. CONCLUSIONS: It is feasible and safe to perform percutaneous DMR with the Biosense system and the channels created with the chosen laser parameters are rapidly sealed with platelet thrombi and at 3 weeks are replaced by well-healed granulation tissue. PMID- 9847989 TI - Non-Alzheimer degenerative dementias. AB - Recent progress in diagnostic criteria of non-Alzheimer degenerative dementias is reviewed. These dementias comprise frontotemporal dementias (including hereditary dementias), primary progressive aphasia and anarthria, corticobasal degeneration, progressive supranuclear palsy and dementia with Lewy bodies. The approach of studying these diseases has changed considerably with genetic and biochemical analyses. A molecular classification is suggested and the clinical significance of this classification is discussed. PMID- 9847990 TI - Memory and language in neurodegenerative diseases. AB - This article reviews recent studies concerning memory and language disorders in Alzheimer's disease and other neurodegenerative conditions. It shows how different memory and language subcomponents may differentially be impaired in different neurodegenerative diseases and at different stages of the same disease. PMID- 9847991 TI - Clinical trials and therapy. AB - Parallel-group studies have established the efficacy of cholinesterase inhibitors in Alzheimer's disease, at least for short-term cognitive and clinical global enhancement. Noncognitive outcomes are currently under scrutiny. Time to reach clinical milestones may be a more useful trial design for new studies aiming to delay appearance or progression of disease. PMID- 9847992 TI - Tubular aggregates in skeletal muscle: their functional significance and mechanisms of pathogenesis. PMID- 9847993 TI - The sarcoglycan complex in limb-girdle muscular dystrophy. AB - The involvement of the sarcoglycan complex in the pathogenesis of muscular dystrophy is becoming increasingly clear. Sarcoglycan gene mutations lead to four forms of autosomal recessive limb-girdle muscular dystrophy. Recent progress has been made with the identification of novel mutations and their correlations with disease. Through this research, a better understanding the molecular pathogenesis of limb-girdle muscular dystrophy has been gained. Finally, animal models are now being used to study viral-mediated gene transfer for the future treatment of this disease. PMID- 9847994 TI - Hereditary inclusion body myopathies. AB - Hereditary inclusion body myopathies comprise autosomal recessive and autosomal dominant muscle disorders that have a variable clinical phenotype but share similar morphological features. These include rimmed vacuoles within muscle fibres and collections of intrasarcoplasmic and intranuclear tubulofilamentous inclusions, 16-18 nm in external diameter. The resemblances and the differences between the sporadic and the hereditary inclusion body myopathies are discussed. Recent advances in the identification of various proteins involved in these diseases are mentioned because they have provided better insight into their underlying pathophysiological mechanisms. Linkage studies have allowed the localization of the genetic defect of some hereditary inclusion body myopathies and related disorders, contributing to their individualization. PMID- 9847995 TI - Ion channel mutations affecting muscle and brain. AB - Voltage-gated ion channels control many aspects of signal transduction in both muscle and nerve. These channels are finely tuned, and either increased or decreased channel activity can adversely affect function. In skeletal muscle, mutations in ion channel genes have been shown to cause both myotonic discharges and episodic paralysis. In cardiac muscle, mutations in related ion channels can produce repolarization defects and fatal arrhythmias. During the past 2 years, a new chapter in the channelopathy story has been opened with the identification of ion channel mutations in the brain that cause disorders ranging from episodic ataxia to epilepsy. PMID- 9847996 TI - The anesthetic myopathies and malignant hyperthermias. AB - Fatal destruction of skeletal muscle coincident with exposure to specific drugs used during anethesia has been recognized as a potentially heritable disorder for more than 30 years. Variable expressivity and incomplete penetrance of the clinical malignant hyperthermia phenotype, together with inherent drawbacks of the in-vitro contracture test confounded efforts to discover the underlying pathogenesis until the application of molecular genetic techniques. On the basis of linkage analysis and mapping of positional candidate genes, mutant alleles at loci on chromosomes 1q, (dihydropyridine-sensitive L-type calcium channel-A1S); 3q, 5p, 7q (dihydropyridine-sensitive L-type calcium channel-LA2), and 19q (ryanodine receptor) are now believed to account for up to 50% of human malignant hyperthermia susceptibility. Although inconsistent genotype-phenotype correlations and doubts regarding the causality of each mutant allele persist, the definition of malignant hyperthermia and relevance of molecular genetic data to the problems of family counseling, population screening, and improved resolution of the malignant hyperthermia phenotype must now be appraised in view of significant locus and allelic genetic heterogeneity. PMID- 9847997 TI - Glycogen storage diseases of muscle. AB - Ten specific enzyme defects of glycogen metabolism affect skeletal muscle alone or in combination with other tissues. The newest addition to this group of disorders is the defect of aldolase A (glycogenosis type XII), a block in terminal glycolysis associated with myopathy and a hemolytic trait. The muscle glycogenoses cause two major syndromes, one characterized by exercise intolerance, cramps, and myoglobinuria, and the other dominated by fixed, often progressive weakness. This review considers sequentially recent advances in the following: clinical features or clinical variants, including a brief description of glycogenosis type XII; animal models, both spontaneous and genetically engineered; physiopathologic mechanisms, especially of the exercise intolerance and myoglobinuria; biochemical and molecular features--molecular defects are just beginning to be discovered for some glycogenoses (e.g. phosphorylase-b-kinase deficiency or branching enzyme deficiency), whereas they form long lists for others, such as acid maltase deficiency and myophosphorylase deficiency; and therapeutic approaches, including enzyme replacement and gene therapy. PMID- 9847998 TI - Fatty acid oxidation defects in muscle. AB - Fatty acid oxidation defects can cause recurrent rhabdomyolysis or chronic progressive muscle weakness. Diagnosis is often possible on blood using tandem mass spectrometry or molecular genetic techniques. Riboflavin and carnitine are effective in some cases of multiple acyl-CoA dehydrogenase deficiency and primary carnitine deficiency, respectively. Controlled trials are needed to evaluate other proposed forms of treatment. PMID- 9847999 TI - Mitochondrial encephalomyopathies. AB - It is nearly a decade since the discovery of the first mutations in mitochondrial DNA associated with mitochondrial encephalomyopathy, and the pace of discovery of new mitochondrial DNA mutations continues unabated. Nuclear gene defects in these disorders have been more difficult to identify; only one is known, but others have been mapped by linkage analysis. The rules governing transmission and segregation of mitochondrial DNA sequence variants are beginning to be unravelled and progress has been made in understanding genotype-phenotype relationships and elucidating mechanisms of pathogenesis. PMID- 9848000 TI - Guillain-Barre syndrome, CIDP and other chronic immune-mediated neuropathies. PMID- 9848001 TI - Painful neuropathies. AB - Pain following peripheral nerve lesion appears to be a paradox because damage of primary afferent nerve fibres carrying nociceptive information should result in hypoalgesia. The very existence of neuropathic pain therefore implies fundamental changes of nociceptive processing and there have been considerable advances in the understanding of factors that precipitate neuropathic pain. This knowledge has already been harnessed for the development of novel analgesic therapies to supplement traditional treatment with anticonvulsant and antidepressants drugs which has shown clear effectiveness in systematic reviews of randomised controlled trials. PMID- 9848002 TI - Metabolic neuropathies. AB - A large epidemiological study has documented that one-third of diabetic patients have peripheral neuropathy. Diabetes duration, poor glycaemic control, smoking and hypertension are all independent predictors of the incidence of diabetic polyneuropathy. High prevalence of autonomic dysfunctions, both sympathetic and parasympathetic, has been found in patients with nonalcoholic chronic liver disease. The pathogenesis of metabolic neuropathy is unclear; even immunologic factors might play a role in the development of diabetic autonomic neuropathy. No specific treatments are available for these neuropathies. Correction of metabolic derangement is fundamental, as shown by the amelioration of peripheral nerve function obtained after successful simultaneous pancreas-kidney transplantation. The therapeuthic potentials of neurotrophins for the prevention and treatment of diabetic neuropathy have to be confirmed in future studies. PMID- 9848003 TI - Autonomic neuropathies. AB - A limited autonomic neuropathy may underlie some unusual clinical syndromes, including the postural tachycardia syndrome, pseudo-obstruction syndrome, heat intolerance, and perhaps chronic fatigue syndrome. Antibodies to autonomic structures are common in diabetes, but their specificity is unknown. The presence of autonomic failure worsens prognosis in the diabetic state. Some autonomic neuropathies are treatable. Familial amyloid polyneuropathy may respond to liver transplantation. There are anecdotal reports of acute panautonomic neuropathy responding to intravenous gamma globulin. Orthostatic hypotension may respond to erythropoietin or midodrine. PMID- 9848004 TI - Tropical myeloneuropathies revisited. AB - An interesting neurological syndrome, characterized by recurrent optic neuritis, cervical myelopathy from syringomyelia, paraparesis, amenorrhea-galactorrhea, and other endocrine problems, has been described among young black women in the French West Indies. The etiology remains unknown, but possible links with Devic's disease, acute disseminated encephalomyelitis, and neurotoxicity from quinolines in Annona muricata teas have been postulated. The largest epidemic of neuropathy in this century occurred in Cuba in 1991-1994. Clinical features and etiologic studies are reviewed. Its primary cause was nutritional. A similar epidemic was recently described in Tanzania. A number of infectious neuropathies and myopathies are reviewed, including leprosy, tuberculosis, hemorrhagic fevers (Ebola and Marburg filoviruses, Lassa, Argentinean and Bolivian arenaviruses), the human retrovirus human T-cell lymphotropic virus type I, Lyme disease and postimmunization neuropathies. The tropics continue to contribute interesting and important clinical conditions that may illuminate the etiopathiogenesis of other common disorders. PMID- 9848005 TI - Inherited demyelinating neuropathies: from gene to disease. AB - Hereditary peripheral neuropathies have traditionally been classified by the clinical disease pattern and mode of inheritance. It only recently became possible to provide a more precise subdivision of the diseases by the discovery of distinct genetic defects. Most inherited peripheral neuropathies are caused by distinct mutations in the genes of three well known myelin components, peripheral myelin protein 22, P0 and the gap junction protein connexin 32. The present review addresses the expression and functional roles of these myelin components, as well as the putative pathomechanisms caused by distinct mutations in the corresponding genes. Moreover, the suitability of mutant animals, such as knock out mice and transgenic rodents, as artificial models for these diseases and their use in the study of possible treatment strategies are discussed. PMID- 9848006 TI - Nerve injury and regeneration: basic insights and therapeutic interventions. AB - Recent observations have provided new insight into neuronal responses to axotomy, signalling of the Schwann cell switch from 'operating' to 'proliferation' mode and temporal molecular changes in the responsiveness of Schwann cells to neuronal signals, as well as into the role of macrophages in Wallerian degeneration, nerve repair and neuropathic pain. Furthermore, promising therapeutic interventions have been developed to promote axon regeneration and to attenuate axotomy-induced neuronal cell death by means of pharmacological treatment or application of neurotrophic proteins using various strategies and routes of delivery. PMID- 9848007 TI - Degenerative diseases. PMID- 9848008 TI - Neuromuscular diseases--muscle. PMID- 9848009 TI - Neuromuscular diseases--nerve. PMID- 9848011 TI - Understanding regulation of cell growth in childhood brain tumors. AB - Advances in the treatment of childhood cancer have come at a remarkable pace, but the search for new treatment strategies remains an urgent one. Brain tumors, the most common childhood solid tumor, are second only to leukemias in overall incidence. In order to develop treatments that improve survival while preserving quality of life for children with brain tumors, a better understanding of brain tumor biology is needed. Tremendous progress has been made in the past several years in the understanding of the machinery of the cell division cycle. This chapter reviews recent insights into the mechanisms controlling cell growth in pediatric brain tumors. PMID- 9848012 TI - Mechanisms of cerebral dysgenesis. AB - Cerebral dysgeneses are increasingly recognized by modern neuroimaging techniques in association with a large variety of pathologies such as epilepsy, learning disabilities, mental retardation, and some psychiatric disorders. Although the mechanisms of these brain dysmorphogeneses are still mostly unknown, the discovery of genes responsible for cerebral dysgeneses and the analysis of relevant animal models have allowed the identification of a few pathophysiologic pathways. This review focuses on some of these recently described molecular mechanisms of cerebral dysgenesis. PMID- 9848013 TI - Understanding childhood language disorders. AB - Developmental language disorders exist in 5% to 10% of preschoolers and have strong genetic implications. There are several variants of dysphasia: mixed receptive/expressive, expressive, or higher order language processing. Preschool children with pervasive developmental disorders are dysphasic as well as autistic. Some undergo a language and behavioral regression, most often as toddlers. The role of subclinical epilepsy in this regression is unknown because it is often ignored. Most dysphasic children speak by schoolage but are at substantial risk for reading/academic difficulty. Powerful new techniques to image the brain during language have no place in the routine workshop of children with dysphasia with or without autism. PMID- 9848014 TI - Pathogenesis of the developmental epilepsies. AB - The developmental epilepsies are distinctive in that they occur in a dynamic and plastic substrate. A variety of acquired insults may present with remarkably similar seizure syndromes that are age-specific and evolve with time. This evolution may be a consequence of the brain injury itself or an alteration of normal brain maturation due to an unfavorable electrical environment. Idiopathic epilepsies are now being correlated with specific gene defects, particularly those involving ion channels and/or neurotransmitter receptors. Despite this specificity, idiopathic seizure syndromes may be caused by mutations in different genes or different mutations in the same gene, and particular syndromes may manifest heterogeneous clinical seizure types. Not all localization-dependent epilepsies are symptomatic, as a growing number of genes have been identified with partial seizure syndromes. Febrile seizures probably do not represent a homogeneous entity, but multiple disorders that may be associated with developmental abnormalities and various coexisting seizure types. PMID- 9848015 TI - Recent advances in developing neuroprotective strategies for perinatal asphyxia. AB - Perinatal asphyxia is the most important cause of acute neurologic injury in the newborn and occurs in approximately six per 1000 term live births. After resuscitation of an infant with birth asphyxia, the emphasis has been on supportive therapy; however, there is increasing evidence that a "therapeutic window" exists in the early hours following the insult, and perhaps for longer, when intervention can attenuate activation of the neurotoxic cascade that leads to delayed cell death hours, days or months later. PMID- 9848016 TI - Allergy, immunology, and related disorders. PMID- 9848017 TI - Genetics of atopic diseases. AB - Atopic (allergic) disorders develop out of a close interaction between genetic predisposition and environmental triggers. A unifying attribute underlying these disorders is atopy, defined as the predisposition of those affected to generate IgE antibodies to environmental antigens and to respond with immediate-type hypersensitivity reactions upon subsequent exposure. Atopy is a heritable trait, and recent studies have identified several genes that engender atopy by increasing either the production of or the responsiveness to IgE. Other genes that contribute to the development of allergic disorders include leukocyte histocompatibility alleles, which specify responsiveness to individual environmental antigens, and disease-related genes, which promote distinctive aspects of an allergic disorder, such as tissue localization. A model is presented whereby the evolution of specific allergic disorders is predicated on the confluence of predisposing genetic elements, coupled with exposure to environmental triggers. PMID- 9848018 TI - Food allergy. AB - The incidence of all allergic diseases appears to be on the increase in industrialized societies. The standard methods of diagnosis (food challenge) and therapy (avoidance) for food allergies remain rather primitive, while the incidence of atopy appears to be increasing. However, a number of advances in recent years hold promise for improvement. This review summarizes important studies and concepts in the field of food allergies. Special emphasis is given to the areas of diagnosis and therapy. PMID- 9848019 TI - Searching for the cause of the increase in asthma. AB - The prevalence of asthma has increased dramatically over the last 30 years, and the clearest evidence for the increase has come from population-based studies of school-aged children and young adults. The strongest established risk factors for asthma are family history and sensitization to one or more indoor allergens. Some studies suggest that the increase in asthma has been part of an overall increase in atopic disease. The American data, however, suggest that hay fever was already common in 1960. Thus the primary increase has been in wheezing among allergic children. It seems unlikely that increases in mite, cockroach, and domestic animal allergens have been sufficient to explain the scale or the consistency of the increase in asthma prevalence. The challenge is to identify a change that could have increased inflammation of the lungs or lowered the threshold for wheezing. PMID- 9848020 TI - Complement deficiency and autoimmunity. AB - Complete deficiency of one of the early components (C1, C4, or C2) of the classical pathway of the complement cascade is one of the strongest genetic risk factors for systemic lupus erythematosus that has been recognized. The lupus that occurs in complement-deficient individuals typically presents in early childhood. The association of complement deficiency and lupus has been known for over two decades, yet the explanation remains somewhat elusive. Complement component deficiencies may be associated with other rheumatic or autoimmune disorders and both partial and acquired complement component deficiencies are also associated with an increased risk of autoimmune disease. This article reviews the current understanding of the relationship between complement component deficiencies and autoimmunity. Recent data from animal models and new types of genetic analyses are reviewed. PMID- 9848021 TI - Tandem spectrometry in newborn screening. AB - Newborn screening for inborn errors of metabolism has improved the diagnosis and treatment of these disorders since the 1960s. Recently tandem mass spectrometry (MS/MS) was developed as a technique for expanding the scope and efficiency of newborn screening for inborn errors. It may offer more efficient identification of phenylketonuria, branched chain ketoaciduria (maple syrup urine disease) and homocystinuria, which are currently screened for by the use of bacterial inhibition assays. MS/MS also identifies analytes characteristic of disorders of fatty acid metabolism and organic acid metabolism, which are not identified in current programs. Recent studies indicate that MS/MS offers the opportunity to expand and advance newborn screening for inborn errors. PMID- 9848022 TI - Clinical and biochemical features of fatty acid oxidation disorders. AB - Inborn errors of fatty acid oxidation (FAO) represent a group of metabolic disorders that has brought forward many interesting developments, as highlighted by the rapid pace of discovery of new defects and by the recognition of an ever increasing spectrum of clinical phenotypes. This review includes a clinical and biochemical summary of the FAO disorders known to date, a synopsis of four recently discovered defects (short-chain 3-hydroxy acyl-CoA [coenzyme A] dehydrogenase deficiency, medium-chain 3-ketoacyl-CoA thiolase deficiency, 3 hydroxy-3-methylglutaryl-CoA synthase deficiency, and long-chain fatty acid transport deficiency) and of two susceptibility variations in the short-chain acyl-CoA dehydrogenase gene, and guidelines for the biochemical work-up of candidate patients. PMID- 9848023 TI - An update on chromosome deletion and microdeletion syndromes. AB - Chromosome deletion and microdeletion syndromes account for an increasing number of clinically recognizable genetic conditions. New deletion syndromes continue to be characterized, and a number of previously described syndromes are being found to be due to chromosomal deletions or microdeletions. Fluorescent in situ hybridization technologies are in wide clinical use to diagnose deletion and microdeletion syndromes, and future uses of these technologies will provide prognostic information for patients and their parents, as the genes responsible for the phenotypic aspects of various deletion syndromes are identified. Future research studies will focus on delineating critical deletion intervals at a molecular level, and identifying candidate genes for the phenotypic features of deletion and microdeletion syndromes, toward the goal of understanding the pathology of the abnormal developmental and physiologic processes involved in each syndrome. PMID- 9848025 TI - Caudal appendage in a full-term infant. PMID- 9848024 TI - Molecular genetics of long-QT syndrome. AB - Recent discoveries of genes involved in long-QT syndrome (LQTS) have led to extensive progress in understanding the molecular basis for this disorder of syncope and sudden cardiac death secondary to ventricular arrhythmias. The emerging unifying theme is that all genes identified to date encode either structural or regulatory subunits for ion channels involved in cardiac repolarization. Defects have been identified in the KCNQ1, HERG, and KCNE1 genes, whose proteins form the K+ channels for the slowly and rapidly inwardly rectifying K+ currents IKs and IKr. Depending on their location and copy number, mutations of KCNQ1 and KCNE1 can cause either autosomal dominant Romano-Ward syndrome or autosomal recessive Jervell and Lange-Nielsen syndrome. The cardiac sodium channel gene, SCN5A, is also mutated in some Romano-Ward cases to produce defects in INa, the cardiac inward Na+ current. The fact that multiple genes are involved and that most LQTS mutations are "private" or "family-specific" complicates molecular diagnosis of LQTS which, currently, is limited to a small number of research laboratories. In future, genotypic determination of LQTS patients and their family members will hopefully lead to improved gene-specific prognostic determinations and therapeutic interventions. PMID- 9848026 TI - Office laboratory procedures, office economics, patient and parent education, and urinary tract infection. AB - This review provides an update on four areas of office practice: office laboratory procedures, office economics, patient and parent education, and urinary tract infection. Thomas Ball reviews physician office laboratories, with updates on the Clinical Laboratory Improvement Amendments, office proficiency testing, and office testing for streptococcal pharyngitis and Helicobacter pylori. Eve Shapiro reports on office economics, focusing on the influence of managed care on pediatric practice. Burris Duncan provides a review of the new National Institutes of Health asthma guidelines, and challenges us to become more involved in patient education. Richard Wahl reviews urinary tract infections, vesicoureteral reflux, dysfunctional voiding, and appropriate imaging studies. Our approach is to provide pediatricians with useful and practical information for their office practices. PMID- 9848027 TI - The problem of HIV/AIDS as related to drug abuse: an introduction. PMID- 9848028 TI - Drug abuse treatment as HIV prevention: changes in social drug use patterns might also reduce risk. AB - Fifty one individuals (37 male and 14 female) were asked to report on the social and behavioral circumstances related to their opiate drug use prior to and during a 90-day methadone detoxification treatment. Data were collected by means of a weekly structured interview. Questions were asked about each occasion of opiate use in the previous week with respect to time, source, cost, social circumstance, etc. Monitored urine samples were tested x 3/week to verify verbal reports. The study demonstrated beneficial effects of the detoxification treatment by showing dramatic decreases in rates and amounts of opiate drug use during treatment. Of perhaps greater significance, large scale changes were also noted in the frequency of use with others. This decline in use with others was most dramatic with respect to strangers and acquaintances. Implications of these observations for HIV transmission are discussed. PMID- 9848029 TI - Stimulant abuse treatment as HIV prevention. AB - Individuals who use illicit stimulants, primarily cocaine and methamphetamine, engage in substantial amounts of HIV-related sexual risk behaviors when under the influence. This paper presents the idea that reductions in stimulant use consequent to drug treatment makes stimulant drug treatment an important HIV prevention tool for this high-risk population. Data are presented to describe many of the HIV-related sexual risks reported by out-of-treatment methamphetamine users and by cocaine and methamphetamine abusers at treatment entry and six months post treatment entry. Overall, our findings demonstrate that following initiation of a treatment episode, stimulant abusers demonstrate significant and relevant reductions in HIV-related sexual behaviors, primarily by reducing the number of sexual partners. Reasons for why stimulant treatment corresponds to HIV transmission risk reductions and suggestions for implementing prevention messages in treatment settings are offered. PMID- 9848030 TI - AIDS risk behavior in opioid dependent patients treated with community reinforcement approach and relationships with psychiatric disorders. AB - This study examined the Community Reinforcement Approach's (CRA) effect on AIDS risk behaviors and the relationship between comorbid psychiatric disorders and the risk for AIDS behavior in opioid dependent patients entering methadone maintenance treatment. Additionally, we looked at AIDS risk behaviors as they related to the Addition Severity Index (ASI), Beck Depression Inventory, Symptom Checklist-90-Revised (SCL-90-R), and the Social Adjustment Scale-Self Report (SAS SR). Subjects (N = 227) were drawn from a large clinical trial that examined the effectiveness of a Community Reinforcement Approach for treatment of opioid dependence. Both CRA and standard treatment demonstrated a significant effect on reduction of AIDS risk behaviors. There was no relationship found regarding comorbid psychiatric disorders with the risk for AIDS behavior. However, there were correlations with other psychiatric, social, and substance abuse variables. Multivariate analyses indicated that increased drug and legal ASI composite scores were the primary predictors of increased AIDS risk behavior. PMID- 9848031 TI - Assessment of HIV risk. AB - Based on the review of existing instruments and analysis of problems encountered in clinical and research practice with one of the most commonly used assessment instruments, the RAB, this paper proposes a number of solutions aimed at improving validity, and efficiency of assessment of HIV risk in drug abusing populations. Briefly, five domains of assessment are discussed: intravenous drug use, high-risk sexual behaviors, knowledge of HIV transmission and methods of prevention, psychological aspects of behavioral change, and epidemiological factors of HIV transmission. The paper discusses also changes in format, scope, and context, as well as scoring procedures that may improve discriminability and sensitivity to detect change of a comprehensive HIV risk assessment instrument. Finally, a process of developing an HIV risk assessment instrument, the ARI-I, which is based on the proposed recommendations and which incorporates methodological improvements discussed in the paper is briefly described. PMID- 9848032 TI - Does intensive outpatient cocaine treatment reduce AIDS risky behaviors? AB - The purposes of this study were: (1) to examine the characteristics of 447 cocaine dependent, first admission outpatients in relation to their AIDS risky behavior at intake; (2) to ascertain whether there was a reduction in risky behavior at follow-up 9 months after admission; and (3) to determine whether reductions in risky behavior were related to patient characteristics, group as compared to individual treatment, or time in treatment. In this sample of cocaine dependent patients entering outpatient treatment, those engaging in higher AIDS risky behaviors were not characterized by any particular demographic profile or by a lack of knowledge about HIV/AIDS. They did have higher scores on the SCL-90 R symptom scales, the Beck Depression Inventory, and higher ratings on the Drug, Alcohol, Family, and Medical scales of the ASI. At 9-month follow-up, AIDS risky behaviors as measured by the RAB were found to have decreased significantly. The degree of improvement was not associated with demographic variables but was predicted by higher intake problem severity and psychological symptomatology scale scores. While improvement in risky behavior was not related to type of treatment or duration of treatment, it was related to decreased substance use. The individuals whose risky behavior decreased were those whose substance use had decreased. Improvement, then required not only being in a treatment program, but also participation and involvement in the program. Treatment and not merely attendance would seem to be critical. PMID- 9848033 TI - Changes in HIV risk behaviors among cocaine-using methadone patients. AB - Cocaine use among methadone patients has been related to higher prevalence of HIV risk behaviors. HIV risk behaviors for cocaine-using patients in methadone treatment (N = 207) were examined for two time periods, the current month in treatment and the month previous to treatment admission. All needle-related and sexually-related risk behaviors (except for needle hygiene) significantly and substantially declined over the average two year time interval. Several variables were associated with needle and sexual risks in multivariate regression analyses. Dropping apparent opiate use underreporters from the analyses did not alter the results. From a harm reduction perspective, high priority should be given to retaining cocaine-using patients in methadone maintenance, intensifying in program services for those with anti-social personality, bipolar disorder or alcoholism, as well as increasing access to needle exchanges and free condoms. PMID- 9848035 TI - The 1998 presidential address the ABA at 30: a SWOT analysis. PMID- 9848034 TI - HIV/AIDS and drug abuse: epidemiology and prevention. AB - In the United States, the AIDS epidemic is a dynamic process with increasing rates of AIDS reported among women, minority populations, heterosexual men, and users of drugs by routes other than injection. The 1993 CDC AIDS definition change has created some difficulties in interpreting trends in the United States. Drug use continues to represent a significant problem among HIV-infected persons. Several strategies have been advanced to decrease transmission of HIV among drug users, their sexual partners and children. However, more effective and comprehensive prevention and treatment strategies are needed. PMID- 9848036 TI - The 1998 Everett Idris Evans Memorial Lecture. "Out of the strong came forth sweetness": on the contribution of military conflicts to the development of burn treatment in Israel. PMID- 9848037 TI - The 1998 Clinical Research Award. Sleep disturbance after burn injury: a frequent yet understudied complication. AB - This study investigated the frequency of sleep disturbance of burn survivors at 3 time points: during hospitalization (time 1: n = 237), 1 week after discharge (time 2: n = 149), and 2 months after discharge (time 3: n = 91). Predictors of sleep disturbance and its relationship to quality of life are explored. Measures of sleep, post-traumatic stress disorder, depression, anxiety, pain, and quality of life were administered at each time point. Fifty percent of participants had sleep disturbance while in the hospital and 1 week after discharge. Forty percent of participants continued to have sleep disturbance 2 months after discharge. In regression equations, emotional distress was a better predictor of sleep disturbance than pain and total body surface area burned at each time point. Sleep disturbance was significantly negatively correlated with all aspects of quality of life represented on the SF-36 Health Survey. Sleep disturbance is a common and often chronic postburn complication that warrants further research. PMID- 9848038 TI - The 1998 Moyer Award. Characteristics of thrombospondin-1 and its cysteine-serine valine-threonine-cysteine-glycine receptor in burn wounds. AB - Thrombospondin-1 (TSP-1), an adhesive glycoprotein, plays an important role in platelet adhesion, inflammation, cell-to-cell interaction, and angiogenesis. TSP 1 is expressed by endothelial cells, fibroblasts, and macrophages. TSP-1's unique cysteine-serine-valine-threonine-cysteine-glycine (CSVTCG) specific receptor plays an important role in the binding and modulation of cellular adhesion and invasion. This article histologically and quantitatively evaluates TSP-1 and its CSVTCG receptor in adult burn wounds over time. Tissue was obtained from burn wounds on several days and samples that were 5 microns thick were placed on slides. Expression of TSP-1 and its CSVTCG receptor were evaluated immunohistochemically and quantitated by computer image analysis in units of absorbance. Immunoglobin G (IgG) (negative) controls were performed and subtracted from the TSP-1 sample to eliminate background absorbance readings. Serum (negative) control was used for the CSVTCG receptor. Platelet concentrates were used as the positive control. A quantitative examination of the results yielded the following information, expressed as absorbance +/- standard error of the mean: TSP-1: day 1, 62.0 +/- 10.13; day 3, 76.2 +/- 6.90; day 5, 36.0 +/- 3.96; day 7, 60.4 +/- 5.67; and day 9, 29.5 +/- 2.91. TSP-1 displays an early peak, followed by a steep decrease over the time period studied. The readings for the CSVTCG receptor are as follows: day 1, 33.8 +/- 1.87; day 3, 34.5 +/- 5.39; day 7, 39.1 +/- 1.93; day 21, 39.1 +/- 1.93; day 28, 34.8 +/- 3.67. In contrast, the CVSTCG receptor continues to be present in the wound over time. Histologic findings are reported, and photographs and a histopathologic analysis are included. The information presented in this article leads to the conclusion that temporal and histologic differences exist in the localization and expression of TSP-1 and its CSVTCG receptor. TSP-1 is up-regulated in injured tissues immediately after the injury; it is rapidly down-regulated as the tissue heals. In contrast, the levels of the CSVTCG receptor remain relatively constant during the healing process. These data are consistent with TSP-1's known role in cell-to cell interaction, including the modulation of the growth factor and protease activity. PMID- 9848039 TI - The 1998 Lindberg Award. Comparison of glycerol preservation with cryopreservation methods on HIV-1 inactivation. AB - Cryopreservation and glycerol preservation are 2 successful methods for long-term preservation of human cadaver skin. Preservation is subjected to strict criteria to minimize the risk of disease transmission. This investigation compares the effects of glycerol preservation and cryopreservation on the inactivation of HIV 1. The effects of glycerol preservation and cryopreservation on inactivation of both extracellular and intracellular HIV-1Ba-L were investigated. After exposing HIV-1Ba-L-infected material to various concentrations of glycerol or to 10% dimethyl sulfoxide followed by cryopreservation, uninfected peripheral blood mononuclear cells were added to the treated material. At different time points during the culture, supernatants were taken to quantify HIV-1Ba-L and reverse transcriptase levels to determine HIV-1Ba-L infectivity. Cell-free HIV-1Ba-L was inactivated within 30 minutes in 70% and 85% glycerol. Also, intracellular HIV 1Ba-L in infected peripheral blood mononuclear cells or infected cadaver skin was completely inactivated by glycerol treatment in vitro. Cryopreservation did not show any extracellular or intracellular HIV-1Ba-L inactivation. Glycerol preservation--but not cryopreservation--of human cadaveric donor skin can inactivate both extracellular and intracellular HIV-1. PMID- 9848040 TI - Chronic indomethacin administration blocks increased body temperature after burn injury in rats. AB - This study investigates the hypothesis that continuous administration of indomethacin (a cyclo-oxygenase inhibitor) will chronically reduce body temperature (TB) in burned rats (i.e., modulate "true" fever). Male Sprague Dawley rats had radio transmitters and osmotic pumps (containing indomethacin) placed in the peritoneal cavity, and 7 days later large full-thickness scald burns were produced. Activity and TB were continually recorded through the 14th postburn day (PBD). There were 4 experimental groups: burn + indomethacin (B-In), n = 9; burn + polyethylene glycol (B-Peg), n = 6; control + indomethacin (C-In), n = 9; and control + polyethylene glycol (C-Peg), n = 6. From PBD 5 through PBD 10, the B-Peg group had consistently and significantly higher TB during light hours than the B-In, C-In, and C-Peg groups. From PBD 7 through PBD 12, the B-In group had an average TB during light hours significantly lower or not different than the C-In and C-Peg groups. These results support the hypothesis that in this burned-rat model chronically increased TB during the light hours may be "fever" (rather than hyperthermia) and that the final effector link could be a prostanoid because changes in activity do not explain the changes in body temperature. PMID- 9848042 TI - A comparison of oral transmucosal fentanyl citrate and oral hydromorphone for inpatient pediatric burn wound care analgesia. AB - The ideal oral wound care analgesic for children should be palatable, provide potent analgesia of rapid onset and short duration, and require minimal, yet appropriate, monitoring. With use of a double-blinded crossover design, we compared the efficacy and safety of oral transmucosal fentanyl citrate (OTFC) (approximately 10 micrograms/kg) with the efficacy and safety of oral hydromorphone (60 micrograms/kg) in 14 pediatric inpatients (ages 4 to 17 years) undergoing daily burn wound care in a ward setting. Pulse oximetry, vital signs, side effects, patient pain scores, and observer scores for cooperation, anxiety, and sedation were recorded. Pulse oximetry, vital signs, cooperation, sedation, incidence of nausea or vomiting, and the amount of time it took to resume normal activities were similar in both treatment groups. OTFC resulted in improved pain scores before wound care and improved anxiolysis during wound care, but at other points it was similar in effect to hydromorphone. We conclude that OTFC is a safe and effective analgesic, that it may provide minor improvements in analgesia and anxiolysis compared with hydromorphone, and that it offers a palatable alternative route of opioid administration without intravenous access for wound care procedures in children. PMID- 9848041 TI - Is the limulus amebocyte lysate the sole predictor of septic episodes in major thermal injuries? AB - Septic episodes in thermal injuries are usually hallmarked by a series of physiologic parameters that include tachypnea, prolonged paralytic ileus, hyperthermia or hypothermia, altered mental status, thrombocytopenia, leukocytosis or unexplained leukopenia, acidosis, and hyperglycemia. Recent studies with polycystic kidney disease have clearly indicated that the limulus amebocyte lysate (LAL) assays were predictive of fungal infections in this patient population. Because both bacteria and fungi produce lipopolysaccharide that can be identified with the LAL assay, we randomly assayed sequential sera of 45 patients with major thermal injuries for positivity in the LAL assay, with use of the QCL-1000 kit (BioWhittaker, Walkersville, Md). The average burn size of this patient population was 63.43% total body surface area. The average age of the patient was 6.2 years. The sex distribution included 30 males and 15 females. The infectious agents included gram-positive cocci and gram-negative rods, and 14 patients had concomitant fungal infections. Eighty-five percent of the patients tested were positive for endotoxin, with levels ranging from < 0.1 EU/mL to > 1.0 EU/mL. The predominant organism isolated before or on the date the serum was drawn was Pseudomonas aeruginosa (51%), followed by Klebsiella pneumoniae (15%). The remaining 34% were a variety of Enterobacteriaceae. Of the 14 patients who yielded a fungus, 3 had negative LAL assays. Two patients with an elevated LAL grew only Staphylococcus epidermidis in the bloodstream and the wounds. These data clearly indicate that the LAL assay cannot be relied on as the sole predictor of septic episodes; however, it can be an adjunctive test to confirm sepsis when the other parameters have been considered. PMID- 9848043 TI - Elder abuse: a call to action. AB - An estimated 2 million people a year are victims of elder abuse, which ranges from neglect and mistreatment to physical abuse. By the year 2020, a full 22% of the population will be aged 65 or older. This demographic explosion demands that we identify and protect those at risk. To investigate the incidence of elder abuse or neglect (EAN) and to determine clinician awareness of associated risk factors, we conducted a 1-year retrospective review of thermally injured patients aged 60 or older. Data included age, total body surface area burned, mechanism of injury, length of hospital stay, mortality, abuse or neglect risk factors, and referral to the appropriate social agency. We found that our elderly patients (n = 28) were poorly screened for EAN. While 64% to 96% of patients were screened for cognitive impairment, overall health, and financial resources, none were screened for risk factors of emotional isolation. None of the patient's caregivers, including any spouses, roommates, or guardians, were screened for risk factors of substance abuse, familial violence, dependency needs, or external stresses. With the use of available data, we were able to place 11 patients on the following levels of abuse or neglect: 1) low risk for abuse; 2) self-neglect; 3) neglect; and 4) abuse. By this scale, 7 patients (64%) were victims of self neglect, 3 patients (27%) were victims of neglect, and 1 patient (9%) was a victim of abuse. Adult Protective Services intervened in 2 cases. Recognizing that all cases of EAN should be preventable, we cannot accept the socioeconomic impact of this entity. The 11 patients identified as victims of neglect, self neglect, or abuse accounted for 135 hospital days and 8 fatalities. Before we can address EAN, health care personnel must be made aware of the problem and routine screening for risk factors must be implemented. The true incidence of EAN is likely underestimated because health care providers have difficulty recognizing its features. A standard assessment tool to screen for neglect or abuse should be used for each older adult admission. PMID- 9848044 TI - Acellular allodermis in burns surgery: 1-year results of a pilot trial. AB - We grafted 10 sites on 6 children with limited (< 25%) areas of body surface available for donor harvest with an acellular allogenic dermis and a thin autograft. Matched control sites were grafted with autograft alone. Study site autografts (0.0074 +/- 0.0007 in, median 0.006 in, range 0.006 to 0.012 in) were thinner than control site autografts (0.0102 +/- 0.0008 in, median 0.012 in, range 0.006 to 0.013 in), with a P value of .015. Endopoints were initial engraftment (in percent) as judged by a blinded experienced observer and Vancouver scar scores. The 6 children (3 girls and 3 boys) had an average age of 5.2 +/- 0.9 years (range 2.8 to 10 years) and an average burn size of 68.7% +/- 6.7% total body surface area (range 47% to 85% total body surface area). The 10 study and control sites were treated with 10 separate procedures; 9 of the procedures were reconstructive and 1 was performed for the excision of an acute burn. Successful initial epithelialization was noted at 7 days postburn for 83% +/- 3.4% (range 60% to 95%) at the cryopreserved acellular human dermis sites and 83.3% +/- 4.3% (range 60% to 98%) at the control sites (not significant, P = .96). At a mean follow-up interval of 43.7 +/- 3.6 weeks (median 52, range 26 to 52 weeks), the differences between the study and control sites in the total mean (pigmentation, vascularity, pliability, and height) of the patients' Vancouver scar scores were not different. PMID- 9848045 TI - A matched-pair, randomized study evaluating the efficacy and safety of Acticoat silver-coated dressing for the treatment of burn wounds. AB - A new silver-coating technology was developed to prevent wound adhesion, limit nosocomial infection, control bacterial growth, and facilitate burn wound care through a silver-coated dressing material. For the purposes of this article, Acticoat (Westaim Biomedical Inc, Fort Saskatchawan, Alberta, Canada) silver coated dressing was used. After in vitro and in vivo studies, a randomized, prospective clinical study was performed to assess the efficacy and ease of use of Acticoat dressing as compared with the efficacy and ease of our institution's standard burn wound care. Thirty burn patients with symmetric wounds were randomized to be treated with either 0.5% silver nitrate solution or Acticoat silver-coated dressing. The dressing was evaluated on the basis of overall patient comfort, ease of use for the wound care provider, and level of antimicrobial effectiveness. Wound pain was rated by the patient using a visual analog scale during dressing removal, application, and 2 hours after application. Ease of use was rated by the nurse providing wound care. Antimicrobial effectiveness was evaluated by quantitative burn wound biopsies performed before and at the end of treatment. Patients found dressing removal less painful with Acticoat than with silver nitrate, but they found the pain to be comparable during application and 2 hours after application. According to the nurses, there was no statistically significant difference in the ease of use. The frequency of burn wound sepsis (> 10(5) organisms per gram of tissue) was less in Acticoat treated wounds than in those treated with silver nitrate (5 vs 16). Secondary bacteremias arising from infected burn wounds were also less frequent with Acticoat than with silver nitrate-treated wounds (1 vs 5). Acticoat dressing offers a new form of dressing for the burn wound, but it requires further investigation with greater numbers of patients in a larger number of centers and in different phases of burn wound care. PMID- 9848046 TI - Nurses' attitudes toward do-not-resuscitate orders. AB - A study was carried out to evaluate burn nurses' attitudes toward do-not resuscitate (DNR) orders. Questionnaires were submitted to 57 staff members, 52 of whom responded. Seventy-five percent of those responding ranged in age from 30 to 49 years. Seventy-one percent were registered nurses or licensed practical nurses. Sixty-five percent of the respondents had been in health care for more than 10 years; 25% had been in burn care for more than 10 years. Fifty percent were Protestants. Thirty-seven percent of the total described themselves as very religious. Ninety-four percent of respondents felt that DNR orders are sometimes appropriate. Eighty-eight percent felt that DNR decisions should not be made solely by the physician. Ninety-five percent felt that input from patients, family members, or both is essential. Having formal ethics committees make such decisions was opposed by 75% of the respondents. Fifty-six percent felt nurses should be involved in such decisions and 21% opposed such involvement. There was considerable uncertainty, disagreement, or both about whether a DNR order should include stopping all medical treatment, ventilators, intravenous fluids, gastric feedings, and medication. The study indicated statistically significant support for the view that DNR orders are appropriate in some cases. Which patients should be given DNR status and who should make the decision about writing DNR orders were more controversial questions. PMID- 9848047 TI - Recombinant human growth hormone decreases lung and liver tissue lipid peroxidation and increases antioxidant activity after thermal injury in rats. AB - The combination of increased oxidant with decreased endogenous polypeptide and protein antioxidant activity corresponds to a decrease in cellular energetics and cell membrane lipid peroxidation. Providing large doses of growth hormone has been shown to preserve cell mass and organ function after burn injury. The role of growth hormone in oxidant injury has not been defined. We determined whether growth hormone altered the degree of lung and liver lipid peroxidation and the activity of glutathione and catalase in lung and liver tissue after burn injury. Four groups of 40 rats each were studied for 48 hours, with 1 group receiving a 20% full-thickness burn, 1 group treated with growth hormone after 20% full thickness burn injury, a control group, and a growth hormone alone-treated group. We found increased lipid peroxidation, measured as malondialdehyde, in lung and liver tissue, and a decrease in glutathione and catalase activities during the 48 hour post-burn period. The addition of growth hormone prevented the lipid peroxidation and significantly increased tissue glutathione and catalase activities with respect to control values. Growth hormone alone also increased endogenous antioxidant levels. We conclude that growth hormone given after burn injury decreases oxidant stress by producing a significant increase in the endogenous antioxidants glutathione and catalase. PMID- 9848048 TI - Angiographic film subtraction using a laser digitizer and computer processing. AB - Digital subtraction angiography has been accepted as an invaluable clinical tool over the past decade; however, film-screen-based angiography is still performed routinely when high-resolution or large field-of-view angiograms are needed. A technique is presented whereby two films from an angiographic sequence are digitized using a high-resolution laser digitizer, and the digitized images are aligned, subtracted, and displayed using the computer. To accommodate for some types of patient motion, an image warping algorithm is presented and discussed in detail. The warping algorithm is piecewise linear, using triangular regions for warping, resulting in a global nonlinear transform across triangle elements. An algorithm describing optimal triangle selection also is discussed. The results show that subtraction images of excellent quality can be produced by the proposed technique, and suggest that, in some settings, digitized subtraction films may be preferred over conventional film subtraction. PMID- 9848049 TI - Perceived fidelity of compressed and reconstructed radiological images: a preliminary exploration of compression, luminance, and viewing distance. AB - The authors' goal was to explore the impact of image compression algorithm and ratio, image luminance, and viewing distance on radiologists' perception of reconstructed image fidelity. Five radiologists viewed 16 sets of four hard-copy chest radiographs prepared for secondary interpretation. Each set included one uncompressed, and three compressed and reconstructed images prepared using three different algorithms but the same compression ratio. The sets were prepared using two subjects, four compression ratios (10:1, 20:1, 30:1, 40:1), and two luminance levels (2,400 cd/m2, standard lightbox illumination, and 200 cd/m2, simulating a typical CRT display). Readers ranked image quality and evaluated obviousness and clinical importance of differences. Viewing distances for image screening, inspection, and comparison were recorded. At 10:1 compression, the compressed and uncompressed images were nearly indistinguishable; the three algorithms were very similar, and differences were rated "not obvious" and "not important." At higher compression, readers consistently preferred uncompressed images, with notable differences between algorithms. The obviousness and clinical importance of differences were rated higher at lightbox luminance. Viewing distances appeared to be idiosyncratic. PMID- 9848050 TI - Clinical evaluation of irreversible data compression for computed radiography of the chest. AB - Efficient data compression is essential for practical daily operation of computed radiography (CR) systems. In this study the clinical applicability of type III irreversible high data compression using an FCR 9501 chest unit (Fuji Photo Film, Tokyo, Japan) was evaluated. Sixty-eight normal and 93 various abnormal cases, with an additional 15 cases of lung cancers with solitary lung nodules, were selected from the file. A pair of hard copies of original images and images reconstructed using type III compression was made for each case. Six radiologists evaluated the image quality by visual rating and receiver operating characteristic (ROC) curve analysis. For all five anatomic regions of normal cases, "original equal to compressed" was the most common response, followed by "original significantly better than compressed." When abnormal cases were evaluated for diagnostic information, there was no significant difference between the compressed and original images. ROC curve analysis on lung nodules with lung cancer showed no significant difference between the two. Compressed CR images using the type III irreversible technique are clinically applicable and acceptable despite slight degradation of image quality. PMID- 9848051 TI - Application of artificial neural networks for quantitative analysis of image data in chest radiographs for detection of interstitial lung disease. AB - The authors have developed an automated computeraided diagnostic (CAD) scheme by using artificial neural networks (ANNs) on quantitative analysis of image data. Three separate ANNs were applied for detection of interstitial disease on digitized chest images. The first ANN was trained with horizontal profiles in regions of interest (ROIs) selected from normal and abnormal chest radiographs for distinguishing between normal and abnormal patterns. For training and testing of the second ANN, the vertical output patterns obtained from the 1st ANN were used for each ROI. The output value of the second ANN was used to distinguish between normal and abnormal ROIs with interstitial infiltrates. If the ratio of the number of abnormal ROIs to the total number of all ROIs in a chest image was greater than a specified threshold level, the image was classified as abnormal. In addition, the third ANN was applied to distinguish between normal and abnormal chest images. The combination of the rule-based method and the third ANN also was applied to the classification between normal and abnormal chest images. The performance of the ANNs was evaluated by means of receiver operating characteristic (ROC) analysis. The average Az value (area under the ROC curve) for distinguishing between normal and abnormal cases was 0.976 +/- 0.012 for 100 chest radiographs that were not used in training of ANNs. The results indicate that the ANN trained with image data can learn some statistical properties associated with interstitial infiltrates in chest radiographs. PMID- 9848052 TI - Contrast limited adaptive histogram equalization image processing to improve the detection of simulated spiculations in dense mammograms. AB - The purpose of this project was to determine whether Contrast Limited Adaptive Histogram Equalization (CLAHE) improves detection of simulated spiculations in dense mammograms. Lines simulating the appearance of spiculations, a common marker of malignancy when visualized with masses, were embedded in dense mammograms digitized at 50 micron pixels, 12 bits deep. Film images with no CLAHE applied were compared to film images with nine different combinations of clip levels and region sizes applied. A simulated spiculation was embedded in a background of dense breast tissue, with the orientation of the spiculation varied. The key variables involved in each trial included the orientation of the spiculation, contrast level of the spiculation and the CLAHE settings applied to the image. Combining the 10 CLAHE conditions, 4 contrast levels and 4 orientations gave 160 combinations. The trials were constructed by pairing 160 combinations of key variables with 40 backgrounds. Twenty student observers were asked to detect the orientation of the spiculation in the image. There was a statistically significant improvement in detection performance for spiculations with CLAHE over unenhanced images when the region size was set at 32 with a clip level of 2, and when the region size was set at 32 with a clip level of 4. The selected CLAHE settings should be tested in the clinic with digital mammograms to determine whether detection of spiculations associated with masses detected at mammography can be improved. PMID- 9848053 TI - Radiology information systems, picture archiving and communication systems, teleradiology--overview and design criteria. AB - Information technology (IT), long taken for granted in commercial settings, is now being utilized for health-care applications. Medical imaging has lagged comparatively due to the extremely vast data content of each frame; thus, the requirement for expensive high-end components. Further, IT in radiology has evolved from two distinctly separate camps--information systems, known as RIS (radiology information systems) and PACS (picture archiving and communications systems). Both RIS and PACS applications have migrated to the PC environment, enabling cost-effective implementation, but from two backgrounds: RIS from vendors using conventional information systems platforms and products, and PACS from radiographic film and modality vendors. The radiology department at Texas Tech University has assembled a seamlessly integrated, enterprise-wide RIS/PACS/teleradiology intranet. The design criteria include user-friendliness, flexibility to respond to changing needs, and open modular architecture to assure interoperability, cost-effectiveness, and future-proofing of investment. Since no single venor could provide an integrated system meeting our specifications, we decided to assume the burden of constructing our own system. As the system integrator, we embrace open architecture, thus enabling the incorporation of industry-standard-compliant, COTS (commercially off the shelf) products as modules. Microsoft Windows NT operating system, Visual C++ programming language, TCP/IP (transmission control protocol/internetworking protocol), relational SQL (structured query language) database, ODBC (open database connectivity), HL-7 (health level seven) and DICOM (digital imaging and communications in medicine) interfaces are utilized. The usage of COTS components reduces the cost to very affordable levels. With this approach, any module in our system can be replaced when outmoded, without affecting other modules in our system, making it truly future-proof. Construction and evolution of our system (TECHRAD) is reviewed. PMID- 9848054 TI - Filmless medical imaging: experiences of the Massachusetts General Hospital. AB - As the concept of picture archival communication systems (PACS) gathers momentum, the vision of a filmless digital department and digital image management has become a reality. This report will discuss the experiences of a major health-care institution with implementation of a large-scale PACS. Specifically, we discuss success with a modular, nonproprietary, multivendor solution that offers flexibility and state of the art functionality at our institution. PMID- 9848055 TI - Picture archiving and communication system activities at the Mayo Clinic Rochester. AB - This report summarizes Mayo Clinic Rochester's experience with a picture archiving and communication system (PACS) to date and discusses the principles that have guided implementation. PMID- 9848056 TI - The primary interpretation workstation: information beyond image data. AB - With the advent of picture archival and communication systems (PACS), the importance of design surrounding primary review workstations has become apparent. To help acceptance of filmless medical imaging, workstations must be developed that serve the needs of both radiologists and referring clinicians. This report will discuss integral requirements of workstation design, including information creation, medical management, medical knowledge, and enabling technologies. PMID- 9848057 TI - Voice recognition--an emerging necessity within radiology: experiences of the Massachusetts General Hospital. AB - Voice recognition represents a technology that is finally ready for prime time use. As radiology services continue to acquire a larger percentage of the shrinking health-care dollar, decreasing operating costs and improved services will become a necessity. The benefits of voice recognition implementation are significant, as are the challenges. This report will discuss the technology, experiences of major health-care institution with implementation, and potential benefits for the radiology practice. PMID- 9848058 TI - Wavelet-based vector quantization for high-fidelity compression and fast transmission of medical images. AB - Compression of medical images has always been viewed with skepticism, since the loss of information involved is thought to affect diagnostic information. However, recent research indicates that some wavelet-based compression techniques may not effectively reduce the image quality, even when subjected to compression ratios up to 30:1. The performance of a recently designed wavelet-based adaptive vector quantization is compared with a well-known wavelet-based scalar quantization technique to demonstrate the superiority of the former technique at compression ratios higher than 30:1. The use of higher compression with high fidelity of the reconstructed images allows fast transmission of images over the Internet for prompt inspection by radiologists at remote locations in an emergency situation, while higher quality images follow in a progressive manner if desired. Such fast and progressive transmission can also be used for downloading large data sets such as the Visible Human at a quality desired by the users for research or education. This new adaptive vector quantization uses a neural networks-based clustering technique for efficient quantization of the wavelet-decomposed subimages, yielding minimal distortion in the reconstructed images undergoing high compression. Results of compression up to 100:1 are shown for 24-bit color and 8-bit monochrome medical images. PMID- 9848059 TI - Hybrid fiber coax networks: a viable option for telemedicine. AB - Cable companies have improved their networks to the degree that they are viable alternatives to currently deployed networks. A detailed look at these cable networks shows them to be superior in bandwidth (capacity), speed, flexibility, and ubiquity. The services that can be provided by these networks are ideal to numerous advanced applications, including telemedicine. PMID- 9848060 TI - A model for achieving total quality-management information technology infrastructure within an integrated delivery system. AB - The strategic development and deployment of a health management information technology infrastructure is discussed from two perspectives for radiologists and for other medical technologists: the integrated delivery system (IDS) perspective and a total quality-management (TQM) perspective. On the one hand, an IDS perspective is important because of the need to prepare radiologists and other medical practitioners to thrive within rapidly changing health organizational models and evolving health service delivery partnership systems. On the other, a TQM perspective is important due to the need to realize an appropriate, efficient, and cost-effective health information infrastructure for developing seamless, integrated radiological and other medical imaging services. Apart from intelligently pursuing an aligned organizational business strategy with the organizational information system strategy, senior radiological managers and medical technologists of health organizations need to pay particular attention to key quality principles for effecting changes in organizational structures and processes to fit changes in information technological requirements, implementations, and innovations. PMID- 9848061 TI - Economic and clinical impact of filmless operation in a multifacility environment. AB - The transition from conventional film-based to filmless operation at the Baltimore Veterans Affairs (VA) Medical Center has resulted in a large number of clinical and economic benefits. The integration of the Department of VA hospitals in Maryland into the VA Maryland Health Care System has resulted in the opportunity to establish a "virtual" radiology and nuclear medicine department. This integrated department is based on a wide area network in which outlying medical centers use a central hospital information system/radiology information system (HIS/RIS) and a central commercial picture archiving and communication system (PACS), as well as a VA-developed image management and communication system. The creation of this virtual radiology/nuclear medicine department has resulted in additional savings and improvements in clinical care. The benefits of the PACS are made possible, to a large extent, by the high level of integration of the PACS and medical modalities with the hospital information and transcription systems. Our experience suggests that it is absolutely essential to integrate the PACS into the patient's electronic medical record to maximize efficiency and clinical effectiveness of the system. PMID- 9848062 TI - [Genetic and molecular diagnostics in retinoblastoma]. AB - Retinoblastoma is a childhood malignancy of the eye. Almost all patients with familial or bilateral disease suffer from the hereditary form of the disease that is caused by germline mutations in one allele of the RB1 gene. Tumor development is initiated by the loss of the second RB allele in a retinal progenitor cell. Most patients with isolated unilateral disease have nonhereditary retinoblastoma and thus do not carry a mutant allele in their germline. In no patient, the presence of a germline mutation can be excluded clinically. Consequently, relatives are at an increased risk for retinoblastoma. Molecular testing, however, enables accurate risk prediction provided that samples are available. In some relatives an increased risk can be excluded by segregation analysis. Most often, however, identification of the disease causing mutation is necessary for accurate risk prediction. Mutation analysis, which is impeded by the size and complexity of the RB gene, is facilitated by use of efficient screening methods. Using these methods, the oncogenic mutation can be identified in most patients. Therefore, predictive testing has become an integral part of contemporary management of retinoblastoma. PMID- 9848063 TI - [Laser-tyndallometry in eyes with acute retinal necrosis syndrome. Erlanger Laser tyndallometry study group]. AB - PURPOSE: Acute retinal necrosis syndrome (ARN) is a rapidly progressive viral necrotic retinitis. We used noninvasive measurement of aqueous flare to quantify blood-ocular barrier breakdown in eyes with ARN und to follow the inflammatory activity and treatment effects. PATIENTS AND METHODS: We report on seven patients (5 males und 2 females, age: 16 to 57 years) with ARN that were treated in our institution between 1991 to 1996. Aqueous flare was quantified regularly during treatment using the laser flare-cell meter (Kowa, FC-1000) following pupillary dilation. RESULTS: Aqueous flare was highly significantly increased in eyes with ARN (145.5 +/- 139.7, range 30 to 367 photon counts/ms). Contralateral, unaffected eyes showed normal flare values (3.6 +/- 0.8 photon counts/ms, p < 0.0001; normals: 4.1 +/- 1.1 photon counts/ms). Flare values correlated with inflammatory activity before and during antiviral treatment. In two patients with ARN caused by herpes simplex-virus aqueous flare values were extremely increased (367 and 316 photon counts/ms), and retinal necrosis was rapidly progressive despite antiviral therapy. One patient developed early ARN in his contralateral eye with flare values increasing from 3.5 to 22.0 photon counts/ms, which quickly normalized following antiviral treatment. CONCLUSIONS: Measurement with the laser flare-cell meter allows quantification of blood-ocular barrier breakdown in eyes with ARN und may be useful for monitoring inflammatory activity during treatment. PMID- 9848064 TI - [Outcomes after penetrating keratoplasty in congenital hereditary corneal endothelial dystrophy (CHED). Report on 13 eyes]. AB - BACKGROUND: Congenital hereditary endothelial dystrophy (CHED) is a rare bilateral corneal disease. The stromal opacity is supposed to result from terminal misdifferentiation of the endothelial cells. In this study we present the morphological and functional results after penetrating keratoplasty in children with CHED who were operated in our department between 1981 and 1997. PATIENTS AND METHODS: In a retrospective clinical cross-sectional study we looked up case histories of 13 eyes from 8 children (7 female, 1 male) with a mean age of 6.0 +/- 3.1 years (ranged from 3 to 14 years). In all children penetrating keratoplasty was performed by one surgeon (GOHN), in 3 eyes using nonmechanical excimer laser trephination. The graft-diameter was in 7 eyes 7.0/7.1 mm, in 2 eyes 7.0/7.2 mm, in 2 eyes 6.5/6.6 mm (resp. 6.8 mm), in 2 eyes 6.0/6.1 mm (resp. 6.2 mm). Fixation of grafts was achieved in 2 eyes by single running suture, in 8 eyes by double running suture and in 3 eyes by multiple interrupted sutures. RESULTS: During a mean follow-up of 4.0 +/- 2.4 years visual acuity increased in all patients (from light perception to 6/20 preoperatively to 2/200 to 14/20 postoperatively). In one patient corneal endothelial-epithelial-decompensation occured (both eyes unterwent previous antiglaucomatous surgery elsewhere), and in 1 patient loosening of one suture happened after 10 month. No immunological graft reaction occurred during follow-up. After excimer laser trephination (3 eyes from 2 patients) visual acuity and corneal astigmatism after surgery was favorable in comparison to all other patients. CONCLUSION: In children with CHED penetrating keratoplasty results not only in a clear cornea but also in a satisfactory functional outcome. Postoperatively periodical morphological controls and assessment of refraction as well as means to prevent amblyopia are indispensable before age 7. PMID- 9848065 TI - [Design and preliminary results of the Erlanger non-high-risk penetrating keratoplasty study]. AB - PURPOSE: The purpose of this prospective study is to examine the non-high-risk penetrating keratoplasty clinically, tyndallometrically and immunological serologically in order to learn more about the pathogenesis, early symptoms and therapy monitoring of corneal graft rejection. PATIENTS AND METHODS: Since february 1997, patients undergoing non-high-risk-PK have been enrolled in this prospective study. Examinations are done preoperatively and in well-defined postoperative gates (6 weeks, 3 months, 6 months, etc.) and include clinical assessments, corneal topography, laser-tyndallometry, corneal endothelial cell count and corneal pachymetry. Preoperatively as well as at each postoperative examination, 10 ml serum and perioperatively corneoscleral donor tissue are collected and frozen (-80 degrees C) to allow immunological and serological examinations in case of graft rejection later. A relational data base (MS Access) guarantees complete and homognous standardized clinical and serological data. We randomised our patients with respect to duration of topical steroid therapy in standard situations (short-time vs. long-time) as well as for systemic steroid therapy (bolus group vs. tapering group) in case of a graft rejection in 2 treatment groups. RESULTS: Up to March 1998, 99 patients have been enrolled in this prospective study. Our report includes the first consecutive 55 patients (25 female, 30 male, recipient age 56.9 +/- 19.4 years, donor age 57.0 +/- 19.9 years). Up to now (follow-up 2.5 +/- 2.1, maximum of 7.3 months) we did not observe immunologic graft rejections. In one patient a primary graft failure occurred. Laser tyndallometry showed a reduction of blood-aqueous barrier breakdown from 6 weeks postoperatively (9.3 +/- 5.7 photon counts/msec) to 3 months postoperatively (6.8 +/- 3.5 photon counts/msec). CONCLUSION: This prospective randomized clinical and immunological study after non-high-risk-PK is supposed to give more information about pathogenesis, early symptoms and therapy monitoring of graft rejection after elective PK. The concept and design of this study are described. Preliminary data may give significant results in a few years. PMID- 9848066 TI - [Cyclocryotherapy in neovascular glaucoma and non-neovascular glaucoma]. AB - BACKGROUND: There are conflicting reports on the value of cyclocryotherapy and it seems that the success rate is depending on glaucoma conditions, the period of follow-up and the technique. This retrospective study was carried out to assess the efficacy and complication rate of cyclocryosurgery for advanced glaucoma with and without neovascularization. PATIENTS AND METHODS: We induced 76 eyes of 75 patients with inadequately controlled glaucoma, which underwent cyclocryotherapy during the period of 1993 and 1996 (treatment time 60 seconds with -80 degrees C, 6-12 applications (mean 9.8 +/- 2.3), 180-360 degree (median 270 degree), diameter of the probe tip 2.5 mm, 1-2 mm distance from the limbus). Depending on the etiology we distinguished between neovascular (NVG) and non-neovascular glaucoma (nNVG). Pre- and postoperative data from all patients were studied retrospectively, for follow-up after 12-36 months patients were examined. RESULTS: Intraocular pressure (IOP) decreased in all patients from 44.7 +/- 12.6 mm Hg preoperatively to 15.6 +/- 6.5 mm Hg postoperatively after a follow-up of 12-36 months. In 88.2% IOP was lowered to < or = 25 mm Hg. NVG showed a mean IOP reduction from 49.1 +/- 12.5 mm Hg before cyclocryotherapy to 15.6 +/- 5.0 mm Hg at follow-up. In the nNVG group IOP was 40.5 +/- 11.3 mm Hg and 15.7 +/- 7.6 mm Hg after cyclocryotherapy. Pressure was controlled (< or = 25 mm Hg) for 83.8% of NVG and 92.3% of nNVG. A cyclocryotherapy-induced intense inflammation was seen more frequent in NVG (43.2%) than in nNVG (17.9%). 2 patients with NVG and 3 with nNVG developed phthisis postoperatively (total 6.7%). CONCLUSIONS: Cyclocryosurgery is an effective method to reduce IOP in advanced, refractory glaucoma, when other methods have failed. The risk/success rate seems to be acceptable. PMID- 9848067 TI - [Computer aided, examiner independent screening for inapparent eye misalignment among 590 infants and preschoolers--a pilot study with an orthoptic gold standard]. AB - BACKGROUND: The purpose of the study was to investigate the computer aided screening method using Purkinje image I and IV reflection patterns for the detection of inapparent eye misalignment and to compare this to an orthoptic examination. MATERIALS AND METHODS: 590 subjects up to 72 months of age with inapparent eye misalignment were recruited from the orthoptic outpatient department and externally. The computer aided screening consisted of taking a series of still video pictures with Purkinje reflection patterns. These were evaluated in an examiner independent way to reach a recommendation whether the child needed an ophthalmological referral or not. As gold standard, an orthoptic examination was performed. For analysis, the data were split by age groups. The orthoptic results were tested for certainty and repeatability. RESULTS: The computer aided examination had the highest sensitivity of 0.82 in the age group up to 2.5 years of age, and a specificity of 0.90. With an estimated prevalence for microtropia of 0.01, the extrapolated positive predictive value was 0.08, and the negative predictive value was 0.998. In the age group up to 2.5 years of age, the percentage of orthoptic examinations without clear result (neither non referral, nor strabismic) was 22.4%, and 6.1% in the screening examination. Among the cases which were examined repeatedly, some were classified as "strabismic" in the beginning, and as "non-referral" in the end in the orthoptic examinations. CONCLUSIONS: The examiner independent, computer aided screening method is a cost effective option for the screening for inapparent eye misalignment, especially in the age group up to 2.5 years of age. If an orthoptic examination was carried out for screening, one should expect a higher rate of false positives, which entails more costly ophthalmological checks. Future studies should assess the validity of the single ortoptic examination as the gold standard in this age group. PMID- 9848068 TI - [Immunohistochemical markers for cytoplasmic antigens in acquired melanosis, malignant melanomas, and nevi of the conjunctiva]. AB - BACKGROUND: Benign and malignant melanocytic lesions of the conjunctiva are difficult to differentiate histologically. At present in the ophthalmologic literature there is not known a high specific and simply applicable histological feature (tumor marker) of differentiation to identify melanocytic lesions of the conjunctiva. METHODS: In this study 55 nevi, 15 primary acquired melanoses (stage Ia) and 54 melanomas of the conjunctiva were examined retrospectively immunhistochemically by antibodies vs. S-100 protein, and the melanoma-associated antigens HMB-45 and NKI/C3 using the labelled avidin-biotin method. 45 patients (25 female, 20 male) with malignant melanomas of the conjunctiva have been followed up clinically. RESULTS: S-100, HMB-45 and NKI/C3 are highly significant markers to identify malignant melanomas of the conjunctiva (sensitivity: S-100: 96.4%, HMB-45: 96.3%, NKI/C3: 98.1%), whereas markers in acquired melanoses (sensitivity: 93.3%, 78.6% and 92.9%), and nevi (sensitivity: 92.9% 40.7%, 98.2%) are not. Positive tumor markers do not correlate with local recurrences or metastasis. The localization of malignant melanomas and the lymphocytic infiltration are not prognostically significant. The only significant risk factor (p = 0.0485) predicting the development of recurrence or metastasis is tumor thickness > 1.5 mm. CONCLUSIONS: The tumor markers S-100, HMB-45 and NKI/C3 cannot differentiate reliably between benign and malignant melanocytic lesions of the conjunctiva, and positive tumor markers do not correlate with local recurrences or metastasis. The only significant risk factor (p = 0.0485) predicting the development of metastasis is tumor thickness (> 1.5 mm). PMID- 9848069 TI - [Ritleng-lacrimal intubation system for injured canaliculi]. AB - BACKGROUND: Intubation of the lacrimal pathway is used to restore the canaliculi. This procedure is often complicated by the so called accordion-phenomenon. PATIENTS AND METHODS: During the last 4 years we used a new system for lacrimal intubation: the Silicon intubationset designed by Ritleng. It consists of a silicon tube which is connected to a polypropylene tube and a stainless probe for lacrimal insertion. We used this set in 24 lacrimal lacerations of the lacrimal pathway. RESULTS: With this lacrimal intubation set lacerated canalicula in 24 patients could be quickly reapproximated in 20 minutes under general anesthesia without complications. CONCLUSION: The silicon intubation set allows quick and safe silicon tubing of injured canalicula. The new intubation set is a great advantage in operating of lacerated canalicula and suitable even for surgeons in training. PMID- 9848070 TI - [Ocular pulse amplitude during manipulation of systemic perfusion parameters]. AB - BACKGROUND: The mechanism for the control of choroidal perfusion is unclear. The present study was designed to investigate for the effect of standardized alteration of systemic perfusion parameters on choroidal hemodynamics in healthy volunteers. METHODS: Intraocular pressure (IOP; German abbreviation: IOD), Ocular Pulse Amplitude (OPA), systemic blood pressure, and pulse rate were measured, and ocular perfusion pressure was calculated in 14 otherwise non-smoking, healthy volunteers prior to and following exercise and smoking. RESULTS: Exercise significantly (sig., p < 0.05) increased pulse rate, systolic blood pressure and ocular perfusion pressure, while it sig. (p < 0.05) reduced IOP and diastolic blood pressure. However OPA was not sig. (p > 0.1) affected by changes in these parameters. Smoking sig. (p < 0.05) increased systolic blood pressure, diastolic blood pressure, pulse rate, and ocular perfusion pressure but did not sig. (p > 0.09) alter OPA. CONCLUSION: Despite affecting ocular and systemic perfusion parameters, exercise and smoking did not alter OPA, suggesting functional isolation i.e. autoregulation of the choroidal and/or ophthalmic artery circulation. PMID- 9848071 TI - [Two contact lens holders for vitreoretinal surgery]. AB - Two new lens holders for contact lens use in vitreoretinal surgery were developed: (1) a flexible hand-fixed lens holder and (2) a lid-retractor-fixed lens holder. Advantages are: better stabilization and centering of the lens, reduction of air bubbles and blood in the interface between cornea and contact lens. PMID- 9848072 TI - [Progressive corneal ectasia after laser in situ keratomileusis (LASIK)]. AB - BACKGROUND: LASIK (Laser in situ keratomileusis) is used in refractive surgery especially for correction of higher degrees of myopia. Preservation of Bowman's layer as well as less postoperative pain and the slight to absent subepithelial haze are regarded as advantages compared to photorefractive keratectomy (PRK). However, numerous serious complications have been described in the literature. PATIENTS AND METHODS: LASIK treatment was performed elsewhere in two patients to treat myopia or myopic astigmatism between -6 and -9 diopters (D). An astigmatism of -6 D was corrected with the LASIK method in another patient with keratoconus. Progressive corneal ectasia of up to seven diopters occurred in all four eyes within a few months. CONCLUSION: Corneal ectasia can occur after LASIK even in low degrees of myopia of less than ten diopters. Recently, -12 D has been specified as the upper limit for this technique. It is especially important to rule out an early keratoconus or a forme fruste of keratoconus preoperatively since keratectasia with particularly rapid progression can occur in such cases: we would like to designate this as "malignant keratoconus". PMID- 9848073 TI - [Subretinal neovascular membranes in membranoproliferative glomerulonephritis type II]. AB - BACKGROUND: So called 'dense deposits' in the retina of a patient suffering from membranoproliferative glomerulonephritis type II (MPGN II) were first demonstrated in 1989. Appearence of subretinal neovascular membranes associated with MPGN II in three patients was described in 1990. PATIENT AND METHODS: We present a 45-year-old male patient, whose insufficiency of the kidney due to a membranoproliferative glomerulonephritis type II required peritoneal dialysis and later transplantation. In both eyes fundus examination revealed typical disseminated, partly confluencing, drusen-like 'dense deposits' and exsudative degeneration of the macula due to subretinal neovascular membranes. Visual acuity was less than 20/400 and visual fields were reduced to small excentrics islands. There was an atrophy of the optic nerve with nearly extinguished VEP in both eyes. CONCLUSION: Flecked changes of the retina called 'dense deposits' associated with membranoproliferative glomerulonephritis type II have to be differentiated in particular from drusen, as well as metabolic-toxic and degenerative retinopathy. Development of subretinal neovascular membranes is attributed to altered retinal pigment epithelium, similar to age-related macular degeneration. An atrophy of the optic nerve associated with this disease has not been described so far. It is possible that vascular damage because of hypertensive changes due to renal dysfunction is the reason for the optic nerve atrophy. With another hypothesis this could be caused by intraocular pressure due to a risen flow of water into the vitreous cavity following the altered osmotic gradient after peritoneal dialysis. PMID- 9848074 TI - [Homologous lamellar central limbokeratoplasty in severe limbal stem cell deficiency]. AB - PURPOSE: It is the purpose of this case report to present a new surgical technique for treatment of severe limbal stem cell deficiency. CASE REPORT: In a 42-year-old female patient with conjunctivalization of the cornea in congenital aniridia a homologous lamellar central limbokeratoplasty was performed. Postoperatively systemic Ciclosporin A was administered in order to prevent the transplanted limbal stem cells from immunological destruction. During the follow up period of 16 months no immune reaction against the transplanted limbal stem cells could be observed. Visual acuity was ameliorated considerably. CONCLUSIONS: Homologous lamellar central limbokeratoplasty is a promising surgical procedure in selected patients with severe limbal stem cell deficiency having less complications compared with penetrating central limbokeratoplasty. PMID- 9848076 TI - Increased levels of advanced glycation endproducts in the lenses and blood vessels of cigarette smokers. AB - BACKGROUND: Advanced glycation endproducts (AGEs) arise from the spontaneous reaction of reducing sugars with the amino groups of macromolecules. AGEs accumulate in tissue as a consequence of diabetes and aging and have been causally implicated in the pathogenesis of several of the end-organ complications of diabetes and aging, including cataract, atherosclerosis, and renal insufficiency. It has been recently proposed that components in mainstream cigarette smoke can react with plasma and extracellular matrix proteins to form covalent adducts with many of the properties of AGEs. We wished to ascertain whether AGEs or immunochemically related molecules are present at higher levels in the tissues of smokers. MATERIALS AND METHODS: Lens and coronary artery specimens from nondiabetic smokers and nondiabetic nonsmokers were examined by immunohistochemistry, immunoelectron microscopy, and ELISA employing several distinct anti-AGE antibodies. In addition, lenticular extracts were tested for AGE-associated fluorescence by fluorescence spectroscopy. RESULTS: Immunoreactive AGEs were present at significantly higher levels in the lenses and lenticular extracts of nondiabetic smokers (p < 0.003). Anti-AGE immunogold staining was diffusely distributed throughout lens fiber cells. AGE-associated fluorescence was significantly increased in the lenticular extracts of nondiabetic smokers (p = 0.005). AGE-immunoreactivity was significantly elevated in coronary arteries from nondiabetic smokers compared with nondiabetic nonsmokers (p = 0.015). CONCLUSIONS: AGEs or immunochemically related molecules are present at higher levels in the tissues of smokers than in nonsmokers, irrespective of diabetes. In view of previous reports implicating AGEs in a causal association with numerous pathologies, these findings have significant ramifications for understanding the etiopathology of diseases associated with smoking, the single greatest preventable cause of morbidity and mortality in the United States. PMID- 9848075 TI - Nitric oxide production and nitric oxide synthase type 2 expression by human mononuclear phagocytes: a review. PMID- 9848078 TI - The mouse formin (Fmn) gene: abundant circular RNA transcripts and gene-targeted deletion analysis. AB - BACKGROUND: Mutations in the mouse formin (Fmn) gene result in limb deformities and incompletely penetrant renal aplasia. A molecular genetic approach was taken to characterize novel circular RNAs from the Fmn gene and to understand the developmental effects of gene-targeted mutations. MATERIALS AND METHODS: RT-PCR and ribonuclease protection analyses were done to characterize the circular RNA transcripts arising from the Fmn gene. Two lines of mice with gene-targeted deletions of specific Fmn exons, namely exon 4 or exon 5, were generated and analyzed. RESULTS: In our analysis of formin cDNAs, we discovered a class of transcripts in which the exon order is reversed such that downstream exons are joined to the acceptor end of a specific exon that lies 5' to them in the genome. RT-PCR and ribonuclease protection analyses indicate that these transcripts are circular and are the major transcripts arising from this locus in adult brain and kidney. To gain insight into the biological function of these transcripts, we have systematically deleted the relevant exons using gene-targeted homologous recombination. The resulting mice fail to produce circular transcripts, but appear to produce normal amounts of the linear RNA isoforms from this locus. While these deficient mice have normal limbs, they display variably penetrant renal aplasia characteristic of other mutant formin alleles. CONCLUSIONS: Our results demonstrate novel circular transcripts arising from the Fmn gene. Moreover, their high levels of expression suggest that they are not products of aberrant splicing events, but instead, may play important biological roles. Mice with gene-targeted deletions of Fmn exons 4 or 5 lack these circular transcripts and have an incompletely penetrant renal agenesis phenotype. While the biologic function of circular Fmn RNA transcripts is not entirely known, our work suggests their possible involvement in kidney development. PMID- 9848080 TI - The Alton Ochsner Medical Foundation. PMID- 9848077 TI - Down-regulation of laminin-5 in breast carcinoma cells. AB - BACKGROUND: Laminin-5 (ln-5), a large heterotrimeric glycoprotein consisting of an alpha 3, beta 3, and gamma 2 chain, is a component of epithelial cell basement membranes that functions as a ligand of the alpha 3 beta 1 and alpha 6 beta 4 integrins to regulate cell adhesion, migration, and morphogenesis. The ln-5 chains show tissue-specific patterns of regulation in tumors derived from different tissues. For example, ln-5 is often up-regulated in gliomas, gastric carcinomas, and squamous carcinomas and down-regulated in prostate and basal cell carcinomas. Ln-5 expression patterns may represent useful tumor markers and help to elucidate the role of ln-5 in tumor progression in different tissue types. MATERIALS AND METHODS: We have studied ln-5 expression patterns in the breast. mRNA levels were examined in tumor and normal breast epithelial cell lines, tissue samples, and immunomagnetically sorted primary cultures using differential display, Northern blotting, and hybridization arrays. Protein levels were examined by immunoprecipitation. Gene integrity was assessed by Southern blotting of representative cell types. RESULTS: Ln-5 alpha 3, beta 3, and gamma 2 mRNA expression was found to be markedly down-regulated in a panel of breast tumor cell lines when compared with normal breast epithelial cells. Ln-5 mRNA was expressed at relatively high levels in MCF-10A immortal normal breast epithelial cells, long-term cultures of normal breast cells, and sorted primary cultures of normal breast luminal epithelial and myoepithelial cells. Reduced, but detectable, levels of ln-5 tended to be expressed in cell lines derived from early-stage breast tumors, whereas expression was generally not detected in cell lines derived from later-stage tumors. In breast tumor tissue specimens, expression of ln alpha 3 and beta 3 mRNAs tended to be reduced relative to levels observed in adjacent nontumor tissue, whereas in gamma 2 levels were elevated in specimens with increased amounts of myoepithelial cells. These ln-5 expression changes could not be attributed to large-scale mutations or gene rearrangements. Ln-5 protein levels were found to reflect mRNA levels in representative cell lines. At senescence, a growth state believed to suppress tumorigenesis, expression of all three ln-5 mRNAs was up-regulated. CONCLUSION: The down regulation of ln-5 mRNA expression in breast tumors cells provides a new molecular marker and suggests that ln-5 functions to control tumor progression in the breast. PMID- 9848079 TI - Decreased fibrinolytic activity in porcine-to-primate cardiac xenotransplantation. AB - BACKGROUND: One major barrier to successful xenotransplantation is acute vascular rejection, a process pathologically characterized by microvascular thrombosis and diffuse fibrin deposition in transplant blood vessels. This pathologic picture may result from a disturbance in the coagulant or fibrinolytic pathways that regulate normal vascular patency. This study evaluated the regulation of fibrinolytic activity defined by tissue plasminogen activator and plasminogen activator inhibitor-1 as it may exist in the setting of acute vascular rejection. MATERIALS AND METHODS, RESULTS: Serial biopsies from cardiac xenotransplants evaluated by immunofluorescence microscopy demonstrated progressive decreases in tissue plasminogen activator and increases in plasminogen activator inhibitor-1. In vitro studies measuring fibrinolytic activity of cell culture medium from porcine aortic endothelial cells stimulated with human serum or autologous porcine serum revealed that human serum triggered as much as 93% increase in antifibrinolytic activity. CONCLUSIONS: These findings demonstrate that porcine vascular endothelial cells change toward an antifibrinolytic state following stimulation with human xenoreactive antibodies and complement. The shift is at least partly explained by an increased ratio of plasminogen activator inhibitor-1 to tissue plasminogen activator, and is at least in part mediated by the activation of complement. This increased antifibrinolytic activity may contribute to the thrombotic diathesis seen in acute vascular rejection in pig-to-primate xenografts. PMID- 9848082 TI - Impaired ovulation in mice with targeted deletion of the neuronal isoform of nitric oxide synthase. AB - BACKGROUND: Nitric oxide (NO) plays an important role in numerous reproductive processes. To date, most studies have assessed the role of NO by using nonspecific pharmacological inhibitors of the precursor to NO, nitric oxide synthase (NOS). These pharmacological NOS inhibitors suppress all isoforms of NOS; thus, the precise contribution of each isoform to female reproductive physiology is unknown. The purpose of this study was to determine the specific role of neuronal NOS (nNOS) in the regulation of ovulation in female mice lacking the gene that encodes for nNOS (nNOS-/-). MATERIALS AND METHODS: Ovulation was assessed in wild-type (WT) and nNOS-/- female mice by examining the number of ovarian rupture sites and number of oocytes recovered from the oviducts following mating or exposure to exogenous gonadotropins (i.e., 5 IU pregnant mares serum gonadotropin [PMSG] and 5 IU human chorionic gonadotropin [hCG]). Ovulatory efficiency was determined as the number of ovulated oocytes per number of ovarian rupture sites. To examine whether ovulatory deficits in nNOS-/- mice were due to alternations in central mechanisms, plasma luteinizing hormone (LH) concentrations were assessed in WT and nNOS-/- mice that were challenged with 25 ng of gonadotropin-releasing hormone (GnRH). To determine whether ovulatory deficits in nNOS-/- mice were due to local ovulation processes, nerves innervating the reproductive tract of WT and nNOS-/- females were examined for the presence of nNOS protein. RESULTS: There were substantial fertility deficits in nNOS-/- female mice; the nNOS-/- mice had fewer oocytes in their oviducts following spontaneous and gonadotropin-stimulated ovulation. Pituitary responsiveness to exogenous GnRH challenge was intact in nNOS-/- mice. Dense nNOS protein staining was observed in nerves innervating the reproductive tracts of WT mice. CONCLUSIONS: The reproductive deficits in nNOS-/- females are most likely due to alternations in the transfer of oocytes from the ovaries to the oviducts during ovulation. These results suggest that defects in neuronally derived NO production may contribute to female infertility. PMID- 9848081 TI - Induction of the chemokine beta peptides, MIP-1 alpha and MIP-1 beta, by lipopolysaccharide is differentially regulated by immunomodulatory cytokines gamma-IFN, IL-10, IL-4, and TGF-beta. AB - The macrophage occupies a central role in the host response to invasion, exerting its control over the developing inflammatory response largely through the elaboration of an assortment of endogenous mediators including many cytokines. The beta chemokine peptides, macrophage inflammatory protein [MIP]-1 alpha and MIP-1 beta, are two such effectors markedly up-regulated in macrophages following exposure to bacterial lipopolysaccharide (LPS). These highly homologous peptides, like the other members of the beta chemokine family, exhibit diverse but partially overlapping biological activity profiles, suggesting that the cellular participants and intensity of an inflammatory response may in part be regulated by selective expression of these chemokines. Studies reported here demonstrate that, in contrast to the "balanced" MIP-1 alpha/MIP-1 beta chemokine responses of LPS-stimulated macrophage cultures in vitro, circulating levels of MIP-1 beta are significantly higher than those of MIP-1 alpha following LPS administration in vivo. Further studies have revealed that several immunomodulatory cytokines known to be up-regulated in vivo as a consequence of exposure to an invasive stimulus (gamma-IFN, IL-10, IL-4, and transforming growth factor [TGF]-beta) down regulated the LPS-induced release of MIP-1 alpha by macrophages in vitro, but spared the MIP-1 beta response. This altered pattern of secretion may explain, at least in part, the high circulating levels of MIP-1 beta relative to MIP-1 alpha observed in vivo in response to LPS challenge. PMID- 9848083 TI - Clioquinol-zinc chelate: a candidate causative agent of subacute myelo-optic neuropathy. AB - BACKGROUND: 5-chloro-7-iodo-8-hydroxyquinoline (clioquinol) was used clinically three decades ago as an oral antiparasitic agent and to increase intestinal absorption of zinc in patients with acrodermatitis enteropathica, a genetic disorder of zinc absorption. Use of clioquinol was epidemiologically linked to subacute myelo-optic neuropathy (SMON), characterized by peripheral neuropathy and blindness, which affected 10,000 patients in Japan. Discontinuation of oral clioquinol use led to elimination of SMON, however, the mechanism of how clioquinol induces neurotoxicity is unclear. MATERIALS AND METHODS: We tested the effect of clioquinol-metal chelates on neural crest-derived melanoma cells. The effect of clioquinol chelates on cells was further studied by electron microscopy and by a mitochondrial potential-sensitive fluorescent dye. RESULTS: Of the ions tested, only clioquinol-zinc chelate demonstrated cytotoxicity. The cytotoxicity of clioquinol-zinc chelate was extremely rapid, suggesting that its primary effect was on the mitochondria. Electron microscopic analysis demonstrated that clioquinol-zinc chelate caused mitochondrial damage. This finding was further confirmed by the observation that clioquinol-zinc chelate caused a decrease in mitochondrial membrane potential. CONCLUSIONS: We demonstrate that clioquinol, in the presence of zinc, is converted to a potent mitochondrial toxin. The phenomenon of clioquinol mediated toxicity appears to be specific to zinc and is not seen with other metals tested. Since clioquinol has been shown to cause increased systemic absorption of zinc in humans, it is likely that clioquinol zinc chelate was present in appreciable levels in patients with SMON and may be the ultimate causative toxin of SMON. PMID- 9848084 TI - No association between the serotonin transporter-linked promoter region polymorphism and either schizophrenia or density of the serotonin transporter in human hippocampus. AB - BACKGROUND: Allelic association case-control analysis of a deletion/insertion polymorphism in the serotonin transporter-linked polymorphic region (5-HTTLPR) has suggested associations with unipolar disorder, bipolar disorder, depression, and Alzheimer's disease. Moreover, the heterozygous long form of the 5-HTTLPR has been associated with increased levels of mRNA for the serotonin transporter (5 HTT) and increased serotonin uptake in lymphoblastic cell lines. This study attempts to determine whether there is an association between 5-HTTLPR genotype and schizophrenia or the binding of [3H]paroxetine to the human hippocampal 5 HTT. MATERIALS AND METHODS: DNA from the cerebellum from 58 schizophrenic and 62 control subjects was used to genotype the 5-HTTLPR. In addition, the relationship between 5-HTTLPR genotype and the affinity and density of [3H]paroxetine binding to the hippocampal 5-HTT was assessed. RESULTS: There were no associations between 5-HTTLPR allotype or genotype and/or the parameters of [3H]paroxetine binding to the hippocampal 5-HTT. CONCLUSIONS: Our data suggest that 5-HTTLPR genotype neither confers an increased susceptibility for schizophrenia nor dictates the expression of the 5-HTT in the human hippocampus. PMID- 9848086 TI - Possible causes of Alzheimer's disease: amyloid fragments, free radicals, and calcium homeostasis. AB - Alzheimer's disease (AD) is a form of dementia in which patients develop neurodegeneration and complete loss of cognitive abilities and die prematurely. No treatment is known for this condition. Evidence points toward beta-amyloid as one of the main causes for cytotoxic processes. The cascade of biochemical events that lead to neuronal death appears to be interference with intracellular calcium homeostasis via activation of calcium channels, intracellular calcium stores, and subsequent production of free radicals by calcium-sensitive enzymes. The glutamatergic system seems to be implicated in mediating the toxic processes. Several strategies promise amelioration of neurodegenerative developments as judging from in vitro experiments. Glutamate receptor-selective drugs, antioxidants, inhibitors of nitric oxide synthase, calcium channel antagonists, receptor or enzyme inhibitors, and growth factors promise help. Especially combinations of drugs that act at different levels might prolong patients' health. PMID- 9848085 TI - Lasp-1, a novel type of actin-binding protein accumulating in cell membrane extensions. AB - The Lasp-1 gene, which has been localized to the q12-q21 region of human chromosome 17, is amplified and overexpressed in human breast cancers. In addition to the previously reported LIM and SH3 domains of Lasp-1, we report here the identification of an actin-binding domain in the core of the protein. This domain is functional as we demonstrate that Lasp-1 binds actin in vivo and in vitro. In addition, confocal analysis of the Lasp-1 subcellular distribution shows that the protein is colocalized with actin at peripheral cell extensions in individual epithelial cancer cells and in transformed fibroblastic cells. Moreover, Lasp-1 is tyrosine phosphorylated in fibroblast cell lines transformed by a constitutively active form of c-Src (c-SrcY527F). Altogether, our results show that Lasp-1 defines a new type of actin-binding protein and suggest that the protein may play a role in a signaling pathway involved in the organization of the cytoskeleton. PMID- 9848087 TI - Local cerebral blood flow during the first hour following acute ligation of multiple arterioles in rat whisker barrel cortex. AB - The objectives are to measure the early time-course of the flows of blood, red cells, and plasma in brain tissue destined to infarct following arterial occlusion. The flux of fluorescent red blood cells (fRBCs) through venules and the arteriovenous transit times (AVTT) of fluorescein-labeled plasma albumin were periodically monitored in anesthetized adult Wistar rats before and up to 60 min after permanent ligations of several small branches of the middle cerebral artery. Of note, fRBC is a function of venular erythrocyte flow and volume, whereas AVTT is a function of plasma flow and volume in visible arteriole capillary-venule units. In another group of anesthetized rats, local cerebral blood flow (ICBF) was measured 1 h after permanent arterial occlusion by [14C]iodoantipyrine (IAP) autoradiography. With this model of focal ischemia, the lesion is highly reproducible and involves part of the whisker barrel cortex. Infarction of this area was observed in 12 of 13 rats. From 10 to 60 min after arterial occlusion, AVTT was nearly four times longer in the ischemic barrel cortex than at the same site before ligations, and fRBC flux was 25%. Neither parameter changed appreciably over this time. After 60 min of ischemia, ICBF on the ipsilateral barrel cortex was 18% of that on the contralateral side and 15% of the sham control value for the same area of the barrel cortex. Since whole blood flow in the ischemic barrel cortex was < 20% of normal at 60 min and AVTT and fRBC flux were essentially constant from 10 to 60 min, the rates of plasma and red cell flows were similarly depressed during the first hour of arteriolar occlusion. In conclusion, such lowering of red cell, plasma, and blood flows produced consistent infarctions in the barrel cortex. PMID- 9848088 TI - Altered hippocampal kainate-receptor mRNA levels in temporal lobe epilepsy patients. AB - This study determined whether hippocampal kainate (KA) receptor mRNA levels were increased or decreased in temporal lobe epilepsy patients compared with nonseizure autopsies. Hippocampal sclerosis (HS; n = 17), nonsclerosis (non-HS; n = 11), and autopsy hippocampi (n = 9) were studied for KA1-2 and GluR5-7 mRNA levels using semiquantitative in situ hybridization techniques, along with neuron densities. Compared with autopsy hippocampi, HS and non-HS cases showed decreased GluR5 and GluR6 hybridization densities per CA2 and/or CA3 pyramid. Furthermore, HS patients demonstrated increased KA2 and GluR5 hybridization densities per granule cell compared with autopsy hippocampi. These findings indicate that chronic temporal lobe seizures were associated with differential changes in hippocampal KA1-2 and GluR5-7 hybridization densities that vary by subfield and pathology group. In temporal lobe epilepsy patients, these results support the hypothesis that pyramidal cell GluR5 and GluR6 mRNA levels are decreased as a consequence of seizures, and in HS patients granule cell KA2 and GluR5 mRNA levels are increased in association with aberrant fascia dentata mossy fiber sprouting and/or hippocampal neuronal loss. PMID- 9848089 TI - Restoration of TEA-induced calcium responses in fibroblasts from Alzheimer's disease patients by a PKC activator. AB - Several alterations in fibroblasts of Alzheimer's disease (AD) patients have been described, including alterations in calcium regulation, protein kinase C (PKC), and potassium (K+) channels. Studies have also found reduced levels of the alpha isoform of PKC in brains and fibroblasts of AD patients. Since PKC is known to regulate ion channels, we studied K+ channel activity in fibroblasts from AD patients in the presence of (2S, 5S)-8-(1-decynyl)benzolactam (BL), a novel activator of PKC with improved selectivity for the alpha, beta, and gamma isoforms. We present evidence for restoration of normal K+ channel function, as measured by TEA-induced [Ca2+]i elevations, due to activation of PKC by BL. Representative patch-clamp data further substantiate the effect of BL on restoration of 113pS K+ channel activity. Immunoblotting analyses using an alpha isozyme-specific PKC antibody confirm that BL-treated fibroblasts of AD patients show increased PKC activation. The present study suggests that PKC activator based restoration of K+ channels may offer another approach to the investigation of AD pathophysiology, which in turn could lead to the development of a useful model for AD therapeutics. PMID- 9848090 TI - Synaptic plasticity in the CA1 area of the hippocampus of scrapie-infected mice. AB - Using conventional in vitro extracellular field potential recordings we have investigated both short- and long-term synaptic plasticity in the hippocampal CA1 area of mice infected with ME7 scrapie. In agreement with earlier studies, no changes were seen in the properties of the Schaffer collateralevoked field excitatory postsynaptic potential during the early stages of the disease (up to 160 days, post inoculation, d.p.i) after which time the recorded potentials were seen to attenuate. Also, up to this time no changes were seen in either paired pulse facilitation or post-tetanic potentiation, which are short-term phenomena associated with brief elevations in presynaptic calcium levels. However, there was a significant shift from the ability of slices to maintain long-term potentiation (LTP) from 100 d.p.i. onwards. In all of these experiments short term potentiation (STP) was preserved, suggesting that from the time that abnormal PrP becomes detectable, or perhaps even earlier, the mechanisms responsible for stabilizing the maintenance phase of LTP are impaired. This result is discussed in terms of the relationship between STP and LTP and how this might be compromised by the conversion of cellular prion protein (PrPC) to the scrapie, protease resistant form of PrP (PrPSc). PMID- 9848091 TI - The diazoxide derivative 7-chloro-3-methyl-3,4-dihydro-2H-1,2,4-benzothiadiazine S,S-dioxide augments AMPA- and GABA-mediated synaptic responses in cultured hippocampal neurons. AB - The diazoxide derivative 7-chloro-3-methyl-3,4-dihydro-2H-1,2,4-benzothiadiazine S,S-dioxide (IDRA21) enhances memory and learning in rodents, most likely by potentiating AMPAergic synaptic activity. We examined IDRA21's effect upon AMPAergic synaptic currents and whole-cell glutamate currents in cultured rat hippocampal neurons to determine whether IDRA21 was a partial modulator of AMPA receptor desensitization and deactivation. Comparable to cyclothiazide, IDRA21 prolonged AMPAergic autaptic currents (5.6 times control, EC50 150 microM) and slowed the rate of AMPA deactivation (3 times control) following 1-ms applications of 1 mM glutamate to excised, outside-out membrane patches. IDRA21 also augmented autaptic GABA currents by 27 +/- 8.1%, although it had two opposing effects, reducing the peak amplitude versus prolonging autaptic GABA currents. IDRA21 (200 microM) inhibited whole-cell GABA currents elicited by exogenously applied 1 mM GABA by 41 +/- 11%. At sufficient concentrations, IDRA21 reduced AMPA receptor desensitization and slowed the rate of deactivation, most consistent with full agonist activity with lower potency compared to cyclothiazide. IDRA21 slightly augments GABAergic synaptic currents. PMID- 9848092 TI - Postherpetic neuralgia: irritable nociceptors and deafferentation. AB - Postherpetic neuralgia (PHN) is a common and often devastatingly painful condition. It is also one of the most extensively investigated of the neuropathic pains. Patients with PHN have been studied using quantitative testing of primary afferent function, skin biopsies, and controlled treatment trials. Together with insights drawn from an extensive and growing literature on experimental models of neuropathic pain these patient studies have provided a preliminary glimpse of the pain-generating mechanisms in PHN. It is clear that both peripheral and central pathophysiological mechanisms contribute to PHN pain. Some PHN patients have abnormal sensitization of unmyelinated cutaneous nociceptors (irritable nociceptors). Such patients characteristically have minimal sensory loss. Other patients have pain associated with small fiber deafferentation. In such patients pain and temperature sensation are profoundly impaired but light moving mechanical stimuli can often produce severe pain (allodynia). In these patients, allodynia may be due to the formation of new connections between nonnociceptive large diameter primary afferents and central pain transmission neurons. Other deafferentation patients have severe spontaneous pain without hyperalgesia or allodynia and presumably have lost both large and small diameter fibers. In this group the pain is likely due to increased spontaneous activity in deafferented central neurons and/or reorganization of central connections. These three types of mechanism may coexist in individual patients and each offers the possibility for developing new therapeutic interventions. PMID- 9848093 TI - Catecholamines potentiate amyloid beta-peptide neurotoxicity: involvement of oxidative stress, mitochondrial dysfunction, and perturbed calcium homeostasis. AB - Oxidative stress and mitochondrial dysfunction are implicated in the neuronal cell death that occurs in physiological settings and in neurodegenerative disorders. In Alzheimer's disease (AD) degenerating neurons are associated with deposits of amyloid beta-peptide (A beta), and there is evidence for increased membrane lipid peroxidation and protein oxidation in the degenerating neurons. Cell culture studies have shown that A beta can disrupt calcium homeostasis and induce apoptosis in neurons by a mechanism involving oxidative stress. We now report that catecholamines (norepinephrine, epinephrine, and dopamine) increase the vulnerability of cultured hippocampal neurons to A beta toxicity. The catecholamines were effective in potentiating A beta toxicity at concentrations of 10-200 microM, with the higher concentrations (100-200 microM) themselves inducing cell death. Serotonin and acetylcholine were not neurotoxic and did not modify A beta toxicity. Levels of membrane lipid peroxidation, and cytoplasmic and mitochondrial reactive oxygen species, were increased following exposure to neurons to A beta, and catecholamines exacerbated the oxidative stress. Subtoxic concentrations of catecholamines exacerbated decreases in mitochondrial energy charge and transmembrane potential caused by A beta, and higher concentrations of catecholamines alone induced mitochondrial dysfunction. Antioxidants (vitamin E, glutathione, and propyl gallate) protected neurons against the damaging effects of A beta and catecholamines, whereas the beta-adrenergic receptor antagonist propanolol and the dopamine (D1) receptor antagonist SCH23390 were ineffective. Measurements of intracellular free Ca2+ ([Ca2+]i) showed that A beta induced a slow elevation of [Ca2+]i which was greatly enhanced in cultures cotreated with catecholamines. Collectively, these data indicate a role for catecholamines in exacerbating A beta-mediated neuronal degeneration in AD and, when taken together with previous findings, suggest roles for oxidative stress induced by catecholamines in several different neurodegenerative conditions. PMID- 9848094 TI - GADD45 is induced in Alzheimer's disease, and protects against apoptosis in vitro. AB - Expression of the growth arrest DNA damage-inducible protein, GADD45, has recently been reported to be induced by a wide range of stimuli, especially those that produce a high level of base pair damage. We have investigated the expression of GADD45 in brain tissue obtained from patients suffering from Alzheimer's disease (AD). Our results demonstrate that many neurons express the GADD45 protein, and that expression of this protein in neurons is associated with expression of the anti-apoptotic protein Bcl-2, and the presence of DNA damage, but not closely associated with tangle-bearing neurons. Additionally, cell lines overexpressing this protein confer resistance to apoptosis induced by DNA damage agent, suggesting that this protein may participate in cell survival mechanisms. PMID- 9848095 TI - Manganese superoxide dismutase overexpression attenuates MPTP toxicity. AB - There is substantial evidence implicating mitochondrial dysfunction and free radical generation in the neurotoxicity of MPTP. Manganese superoxide dismutase (MnSOD) is the primary antioxidant enzyme protecting against superoxide radicals produced within mitochondria. Overexpression of human MnSOD in transgenic mice resulted in increased MnSOD localized to mitochondria in neurons and a 50% increase in total MnSOD activity in brain homogenates. We found that MPTP toxicity was significantly attenuated in the MnSOD transgenic mice which overexpress the human manganese superoxide dismutase gene, with these mice showing threefold greater dopamine levels than controls following MPTP. There were no alterations in MPP+ levels, suggesting that the effects were not due to altered metabolism of MPTP. A significant increase in 3-nitrotyrosine levels was seen in littermate controls but not in transgenic mice overexpressing human MnSOD. These results provide further evidence implicating mitochondrial dysfunction and oxidative damage in the pathogenesis of MPTP neurotoxicity. PMID- 9848097 TI - Uptake and internalization of exogenous apolipoprotein E3 by cultured human central nervous system neurons. AB - Apolipoprotein E (apoE) has been confirmed as a risk factor for late-onset Alzheimer's disease (AD) and is associated with neurofibrillary tangles and senile plaques, the microscopic pathological characteristics of AD. There has been no direct evidence that human central nervous system neurons can take up and internalize exogenous apoE, which may be important in order for apoE to be involved in the development of the disease. This paper demonstrates by immunohistochemistry and confocal microscopy that cultured human brain neurons can take up and internalize exogenous recombinant human apoE3. We confirm that neurons express the low-density lipoprotein receptor-related protein (LRP) but do not express the low-density lipoprotein receptor. We also demonstrate that the LRP mediates the neuronal uptake of apoE. PMID- 9848096 TI - Cloning and expression of a P2y purinoceptor from the adult bovine corpus callosum. AB - We have isolated an ATP receptor clone by screening a bovine corpus callosum cDNA library. The clone includes one open reading frame encoding for a protein of 373 amino acid residues (42 kDa) which belongs to the G-protein-coupled receptor superfamily. In Xenopus oocytes, this clone expressed an ATP receptor that triggered an oscillatory current in response to ATP (EC50 approximately 20 microM). This current may have resulted from the activation of phospholipase C, the formation of inositol trisphosphate, and the release of Ca2+, which then opens Cl- channels. The order of potency for ATP receptor agonists was 2-MeSATP approximately ATP >> alpha, beta-MeATP > adenosine, and UTP was ineffective, a pharmacological profile consistent with that of a P2y purinoceptor. Northern blot analysis of mRNAs from various bovine brain tissues showed that the gene is expressed in the cerebellum, medulla, corpus callosum, hippocampus, superior colliculus, frontal cortex, and retina. In situ RT-PCR showed transcripts of the gene in many glial cells and endothelial cells of the corpus callosum. The cloned receptor may play an important role in neuron-glial signaling under normal and pathological conditions. PMID- 9848098 TI - Flemish and Dutch mutations in amyloid beta precursor protein have different effects on amyloid beta secretion. AB - Mutations in the amyloid beta precursor protein (APP) gene cosegregate with autosomal dominant Alzheimer disease (AD). Brain pathology of AD is characterized by amyloid deposition in senile plaques and by neurofibrillary tangles. Amyloid deposits in AD brains consist of amyloid beta (A beta), a 4-kDa proteolytic product of APP. In contrast, two other mutations in APP, the Flemish APP692 and Dutch APP693 mutations, are associated with autosomal dominant cerebral hemorrhages due to congophilic amyloid angiopathy (CAA) in the presence or absence of AD pathology, respectively. Both mutations are located within A beta near the constitutive cleavage site. While a common effect of AD-linked mutations is to elevate A beta 42 extracellular concentrations, not much is known about the effect of APP692 and APP693. Here we provide evidence that APP692 and APP693 have a different effect on A beta secretion as determined by cDNA transfection experiments. While APP692 upregulates both A beta 40 and A beta 42 secretion, APP693 does not. These data corroborate with previous findings that increased A beta secretion and particularly of A beta 42, is specific for AD pathology. PMID- 9848099 TI - Effect of 4-[3,5-bis(trimethylsilyl)benzamido] benzoic acid (TAC-101) on the liver metastasis of colon 26-L5 carcinoma cells. AB - We found that oral administration of the benzoic acid derivative, TAC-101 ?4-[3,5 bis(trimethylsilyl)benzamido] benzoic acid? significantly inhibited experimental liver metastasis of murine colon 26-L5 carcinoma cells, whereas all-trans retinoic acid (ATRA) did not show any inhibitory effect. Treatment with more than 10 microM TAC-101 for 24 h showed direct cytotoxicity against tumor cells in vitro. In contrast, ATRA did not have any direct cytotoxicity. TAC-101 also inhibited the tumor cell invasion enhanced by TPA (12-O-tetradecanoylphorbol-13 acetate; AP-1 activator) in a concentration-dependent manner, whereas ATRA did not. Furthermore, zymographic analysis revealed that noncytotoxic concentrations (< 10 microM) of TAC-101 inhibited TPA-induced production of urokinase-type plasminogen activator (u-PA) and matrix metalloproteinase (MMP)-9 from tumor cells, which is considered to be associated with their invasive and metastatic potentials. These results suggest that such an inhibitory effect is partly due to the ability of TAC-101 to bind a retinoic acid receptor (RAR)-alpha and consequently inhibit metastasis-related gene transcription by interfering with AP 1/DNA binding, as we showed previously. On the other hand, TAC-101 also inhibited the production of MMP-2, which is not affected by TPA. Therefore, the antimetastatic effect of TAC-101 includes an alternative regulatory mechanism for MMP production. These results indicate that the in vivo antimetastatic effect of TAC-101 is partly due to the cytotoxicity against tumor cells, which may be caused by the induction of apoptosis, and inhibition of the production of invasion-associated proteolytic enzymes. PMID- 9848100 TI - Differential effect of interleukin 10 on interleukin 12- and interleukin 2 mediated killer induction from blood mononuclear cells. AB - We investigated the regulatory effect of interleukin (IL) 10 on IL-12-inducible killer activity of blood mononuclear cells (MNC) of healthy subjects. IL-10 suppressed non-MHC-restricted killer activity induced by IL-12 in a dose dependent manner. Cytotoxicity against Daudi cells of blood MNC activated with optimal dose (100 U/ml) of IL-12 was significantly suppressed from 36.6 +/- 6.8% (mean +/- SE) to 26.0 +/- 5.0% by 100 ng/ml of IL-10 (n = 5; P < 0.05) at an E/T ratio of 20. IL-10, however, did not suppress killer induction by IL-2 from blood MNC. On the other hand, IL-4, a known inhibitor of IL-2-mediated killer induction, had no effect on IL-12-mediated killer induction from blood MNC. Moreover, production of interferon-gamma by IL-12-stimulated MNC was suppressed by IL-10. Because the presence of IL-10 is reported in the tumor-growing site, these observations suggest that IL-10 acts as a key cytokine in the suppression of IL-12-mediated antitumor response in situ. PMID- 9848101 TI - Spatial and temporal expression of angiogenic molecules during tumor growth and progression. AB - The growth and metastasis of cancer directly correlates with tumor angiogenesis. A better understanding of the expression of regulatory factors controlling angiogenesis is important in exploiting this process therapeutically. Our present study demonstrates that small tumors (3-4 mm in diameter) express more basic fibroblast growth factor (bFGF) and interleukin 8 (IL-8) than large tumors (> 10 mm in diameter), whereas more vascular endothelial growth factor (VEGF) is expressed in large tumors. Immunostaining showed a heterogeneous distribution of angiogenic factors within the tumor; expression of bFGF and IL-8 was highest on the periphery of a large tumor, where cell division is maximum. VEGF expression was higher in the center of the tumor. In vitro studies demonstrated that sparse cultures of tumor cells expressed higher levels of bFGF and IL-8 than confluent cultures. In contrast, the expression of bFGF and IL-8 was not diminished in tumor cells growing on confluent monolayers of normal cells. VEGF expression was upregulated by cell density irrespective of contact with tumor cells or normal cells. These results demonstrate that the expression of different angiogenic factors in tumor cells can be regulated by their proximity to other tumor cells or host cells. PMID- 9848103 TI - Tamoxifen downregulates signal transduction and is synergistic with tiazofurin in human breast carcinoma MDA-MB-435 cells. AB - Breast carcinoma is a leading cause of cancer death in women in the US. Tamoxifen (TAM), an antiestrogen, is used as a chemopreventive and chemotherapeutic compound against human breast carcinoma. Tiazofurin (TR), an oncolytic C nucleoside, inhibits IMP dehydrogenase activity, decreases cellular GTP pools, and downregulates ras gene expression. MDA-MB-435 cells are estrogen receptor negative human breast carcinoma cells that have elevated signal transduction activity. Because TR and TAM decrease signal transduction enzyme activity and inositol 1,4,5-trisphosphate (IP3) concentration via different mechanisms, we tested the hypothesis that the two compounds may be synergistic in human breast carcinoma cells. In MDA-MB-435 cells in growth inhibition assay, the IC50s for TR and TAM were (mean +/- SE) 17 +/- 1.2 and 12 +/- 1.1 microM; in clonogenic assays they were 4 +/- 0.3 and 0.7 +/- 0.3 microM, respectively. When TR was added to MDA-MB-435 cells, followed 12 h later by TAM, synergism was observed in growth inhibition and clonogenic assays and in the reduction of IP3 concentration. The latter may explain, at least in part, the synergistic action of TR and TAM in these cells. The synergistic action of TR and TAM may have implication in the clinical treatment of human breast carcinoma. PMID- 9848104 TI - Sodium pump and Na+/H+ antiport restoration in erythrocytes from cancer patients in remission. AB - We previously showed that in erythrocytes from cancer patients, the sodium pump is decreased and the optimal intracellular pH for Na+/H+ antiport activity is shifted toward an acidic value. We now have studied these sodium transporters in erythrocytes from patients in remission. Moreover, we intended to explain why the transporters were impaired in erythrocytes, which have no apparent bearing on cancer tissues. The sodium pump was studied through a microcalorimetric method, and the Na+/H+ antiport by a titrimetric method. In patients in remission the sodium pump activity returned to normal: 15.10 +/- 6.00 vs. 14.12 +/- 5.28 mW/l cells for remission and control, respectively. The optimal intracellular pH for Na+/H+ antiport activity was identical in remission and control: 6.09 +/- 0.23 vs. 6.10 +/- 0.10. Restoration of sodium pump activity and optimal intracellular pH for Na+/H+ antiport activity in erythrocytes were thus linked to remission. Moreover, we showed that the impairments of the sodium transporters were due to the presence of plasma-borne factors, the existence of which explained why the sodium transporters were impaired in erythrocytes. PMID- 9848102 TI - Overexpression of bcl-2 or bcl-XL fails to inhibit apoptosis mediated by a novel retinoid. AB - Overexpression of bcl-2 or bcl-XL has been found to inhibit the induction of apoptosis in malignant cells by a large number of agents including a wide variety of chemotherapeutic drugs. CD437 ?6-[3-(1-adamantyl)-4 hydroxyphenyl]-2 naphthalene carboxylic acid? is a novel retinoid that induces apoptosis in a number of malignant cells through a unique mechanism of action. The addition of 1 microM CD437 to HL-60/NEO cells resulted in capase 3 (CPP32) activation and poly(ADP-ribose) polymerase (PARP) cleavage in 3 h whereas in bcl-2- or bcl-XL overexpressing HL-60 cells CD437 induced CPP32 activation and PARP cleavage in 6 h. Although 50 and 300 nM CD437 were required to induce PARP cleavage in HL 60/NEO and HL-60/bcl-2, HL-60/bcl-XL cells, respectively, maximal apoptosis in both cell lines was achieved utilizing 300 nM CD437. All three cell lines, however, share identical dose-response curves in terms of their growth inhibition, suggesting that CD437-mediated inhibition of growth and induction of apoptosis represent two distinct and separable processes. In addition, CD437 induces GI arrest as well as p21WAFI/CIPI mRNA expression in these cells despite the overexpression of bcl-2 or bcl-XL. CD437 induced mitochondrial instability as indicated by cytochrome c leakage into the cytoplasm in all three cell lines. CD437 also induced growth inhibition and apoptosis of an apoptosis-resistant variant of the HL-60 cell line (HCW-2), which switched expression from bcl-2 to bcl-XL. CD437-mediated apoptosis is not accompanied by downregulation of bcl-2 or bcl-XL or upregulation of bax. The reason for the inability of bcl-2 or bcl-XL overexpression to inhibit CD437-mediated apoptosis is unclear. The ability of CD437 to initiate apoptosis in a spectrum of malignant cells without interference from bcl-2 or bcl-XL overexpression suggests that CD437 may possess significant therapeutic potential in the treatment of malignancy. PMID- 9848105 TI - Spontaneous neoplasms in control Wistar rats: a comparison of reviews. AB - Spontaneous neoplasms in 930 control Wistar rats from five carcinogenicity bioassays conducted between 1990 and 1995 were reviewed and compared with review findings in studies between 1980 and 1990. Mean survival at 104 weeks was 55% for males and 60% for females, similar to that of the previous review. A total of 1599 neoplasms was diagnosed in 361 (78%) male and 415 (89%) female rats; 1293 (81%) of these were benign and 306 (19%) were malignant (11% with metastases). Sixty-eight percent of all neoplasms were in endocrine and integumentary systems, similar to 74% seen in the previous review. Most common neoplasms (affecting > 7% of either sex) were pituitary adenoma (34% of males, 50% of females), benign adrenal pheochromocytoma (10% of males, 1% of females), thyroid C cell adenoma (6% of males, 8% of females), mammary fibroadenoma (3% of males, 36% of females), keratoacanthoma (11% of males, 0.6% of females), testicular interstitial cell tumor (11% of males), uterine stromal polyp (16% of females), pancreatic acinar cell adenoma (13% of males, 0.6% of females), and benign thymoma (3% of males, 8% of females). Seventeen neoplasms affecting 2 to 6.9% of either sex included adrenal cortical adenoma, thyroid follicular adenoma, pancreatic islet cell adenoma, pituitary carcinoma, mammary adenoma, mammary adenocarcinoma, fibroma, fibrosarcoma, dermal papilloma, uterine schwannoma, uterine granular cell tumor, pancreatic acinar cell carcinoma, hepatocellular adenoma, lymphoma, granular cell meningioma, renal mesenchymal tumor, and hemangiosarcoma. Remaining neoplasms occurred in fewer than 2% of animals. Mean tumor incidence did not differ significantly between our two reviews. Ratios of benign to malignant neoplasms were similar in both reviews and percentages of survival at 104 weeks were similar. Between the two reviews, greater than threefold increase in frequency of some neoplasms occurred only in males and included keratoacanthomas, pancreatic acinar cell adenomas/carcinomas, and astrocytomas. Frequencies of remaining neoplasms were within twofold or within 10% of previous frequencies. Some neoplasms diagnosed in this review but not in the previous review included cardiac schwannoma, pilomatrixoma, parathyroid adenoma, and prostatic adenoma but incidence was approximately 1% for any one tumor. Based on these reviews, Wistar rats appear to have a predilection to pituitary neoplasms and mammary fibroadenomas (females). PMID- 9848106 TI - Symposium overview: chemical modulation of neuroreceptors and channels via intracellular components. AB - Whereas the roles of G proteins and protein kinases in various neuroreceptors and ion channels have been studied extensively, their roles in the actions of drugs and toxicants on these receptors and channels remain to be elucidated. Almost all drugs and toxicants exert multiple actions on multiple target sites, and there is no reason to assume that a chemical modulates a receptor/channel via a single mechanism. In fact, experimental evidence is slowly but steadily being accumulated to indicate that certain drugs and toxicants modulate neuroreceptor/channel functions through interactions with intracellular components such as G proteins and protein kinases. Multiple actions of a toxicant on various receptors/channels may be explained on the basis of its interaction with the G protein/kinase system that is a common denominator of the target sites. This is a virgin field that promises a quantum leap in the coming years. Each presentation and discussion will focus on expected future developments and potential significance in the field of neurotoxicology. PMID- 9848107 TI - Role of kinetics in acute lethality of nonreactive volatile organic compounds (VOCs). AB - The role of kinetics in the acute inhalation toxicity of nonreactive, volatile organic compounds (VOCs), including lipophilic and hydrophilic compounds, was analyzed with a physiologically based pharmacokinetic (PB-PK) model for the rat. For 15 VOCs, a total of 23 LC50 values were retrieved from the literature. It was observed that the external exposure parameter (LC50.exposure length; in ppm.h), varied approximately 60-fold. Concentrations of compounds in the lipoid brain fraction were simulated using a kinetic model. This lead to a more than 10-fold reduction in the toxic range of the 15 VOCs. The average value for this simulated dose surrogate was 70 +/- 31 mM for all VOCs. These observations support the presumption that nonspecific, acute narcotic lethality is directly related to the extent of VOC distribution into lipoid brain constituents. The present results can be used for estimation of the acute lethality of nonreactive VOCs on the basis of kinetic simulations. In addition, the presently calculated dose surrogate for VOC lethality in rats is found to be very similar to the reported internal lethal concentrations of so-called "baseline toxicity compounds" in fish. This indicates a common mechanism of acute VOC toxicity among mammalian and aquatic species. PMID- 9848108 TI - The extent of dichloroacetate formation from trichloroethylene, chloral hydrate, trichloroacetate, and trichloroethanol in B6C3F1 mice. AB - Conflicting data have been published related to the formation of dichloroacetate (DCA) from trichloroethylene (TRI), chloral hydrate (CH), or trichloroacetic acid (TCA) in B6C3F1 mice. TCA is usually indicated as the primary metabolic precursor to DCA. Model simulations based on the known pharmacokinetics of TCA and DCA predicted blood concentrations of DCA that were 10- to 100-fold lower than previously published reports. Because DCA has also been shown to form as an artifact during sample processing, we reevaluated the source of the reported DCA, i.e., whether it was metabolically derived or formed as an artifact. Male B6C3F1 mice were dosed with TRI, CH, trichloroethanol (TCE), or TCA and metabolic profiles of each were determined. DCA was not detected in any of these samples above the assay LOQ of 1.9 microM of whole blood. In order to slow the clearance of DCA, mice were pretreated for 2 weeks with 2 g/liter of DCA in their drinking water. Even under this pretreatment condition, no DCA was detected from a 100 mg/kg i.v. dose of TCA. Although there is significant uncertainty in the amount of DCA that could be generated from TRI or its metabolites, our experimental data and pharmacokinetic model simulations suggest that DCA is likely formed as a short-lived intermediate metabolite. However, its rapid elimination relative to its formation from TCA prevents the accumulation of measurable amounts of DCA in the blood. PMID- 9848109 TI - In vivo percutaneous absorption of boric acid, borax, and disodium octaborate tetrahydrate in humans compared to in vitro absorption in human skin from infinite and finite doses. AB - Literature from the first half of this century report concern for toxicity from topical use of boric acid, but assessment of percutaneous absorption has been impaired by lack of analytical sensitivity. Analytical methods in this study included inductively coupled plasma-mass spectrometry which now allows quantitation of percutaneous absorption of 10B in 10B-enriched boric acid, borax, and disodium octaborate tetrahydrate (DOT) in biological matrices. This made it possible, in the presence of comparatively large natural dietary boron intakes for the in vivo segment of this study, to quantify the boron passing through skin. Human volunteers were dosed with 10B-enriched boric acid, 5.0%, borax, 5.0%, or disodium octaborate tetrahydrate, 10%, in aqueous solutions. Urinalysis, for boron and changes in boron isotope ratios, was used to measure absorption. Boric acid in vivo percutaneous absorption was 0.226 (SD = 0.125) mean percentage dose, with flux and permeability constant (Kp) calculated at 0.009 microgram/cm2/h and 1.9 x 10(-7) cm/h, respectively. Borax absorption was 0.210 (SD = 0.194) mean percentage of dose, with flux and Kp calculated at 0.009 microgram/cm2/h and 1.8 x 10(-7) cm/h, respectively. DOT absorption was 0.122 (SD = 0.108) mean percentage, with flux and Kp calculated at 0.01 microgram/cm2/h and 1.0 x 10(-7) cm/h, respectively. Pretreatment with the potential skin irritant 2% sodium lauryl sulfate had no effect on boron skin absorption. In vitro human skin percentage of doses of boric acid absorbed were 1.2 for a 0.05% solution, 0.28 for a 0.5% solution, and 0.70 for a 5.0% solution. These absorption amounts translated into flux values of, respectively, 0.25, 0.58, and 14.58 micrograms/cm2/h and permeability constants (Kp) of 5.0 x 10(-4), 1.2 x 10(-4), and 2.9 x 10(-4) cm/h for the 0.05, 0.5, and 5.0% solutions. The above in vitro doses were at infinite, 1000 microliters/cm2 volume. At 2 microliters/cm2 (the in vivo dosing volume), flux decreased some 200-fold to 0.07 microgram/cm2/h and Kp of 1.4 x 10(-6) cm/h, while percentage of dose absorbed was 1.75%. Borax dosed at 5.0%/1000 microliters/cm2 had 0.41% dose absorbed, flux at 8.5 micrograms/cm2/h, and Kp was 1.7 x 10(-4) cm/h. Disodium octaborate tetrahydrate (DOT) dosed at 10%/1000 microliters/cm2 was 0.19% dose absorbed, flux at 7.9 micrograms/cm2/h, and Kp was 0.8 x 10(-4) cm/h. These in vitro results from infinite doses (1000 microliters/cm2) were 1000-fold greater than those obtained in the companion in vivo study. The results from the finite (2 microliters/cm2) dosing were closer (10-fold difference) to the in vivo results. General application of infinite dose percutaneous absorption values for risk assessment is questioned by these results. These in vivo results show that percutaneous absorption of boron, as boric acid, borax, and disodium octaborate tetrahydrate, through intact human skin, is low and is significantly less than the average daily dietary intake. This very low boron skin absorption makes it apparent that, for the borates tested, the use of gloves to prevent systemic uptake is unnecessary. These findings do not apply to abraded or otherwise damaged skin. PMID- 9848110 TI - Glucuronidation of catechol estrogens by expressed human UDP glucuronosyltransferases (UGTs) 1A1, 1A3, and 2B7. AB - Catechol estrogens are major estrogen metabolites in mammals and are the most potent naturally occurring inhibitors of catecholamine metabolism. These estrogen compounds have been implicated in carcinogenic activity and the 4/2 hydroxyestradiol concentration has been shown to be elevated in neoplastic human mammary tissue compared to normal human breast tissue. Three human liver UDP glucuronosyltransferases, UGT2B7, UGT1A1, and UGT1A3, have been shown to catalyze the glucuronidation of catechol estrogens and lead to their enhanced elimination via urine or bile. The present study was designed to study the kinetic interaction of expressed human UGT2B7(Y) or (H), UGT1A1, and UGT1A3 toward 2- and 4-hydroxycatechol estrogens. cDNAs encoding UGT2B7(Y) or (H), UGT1A1, and UGT1A3 were expressed in HK293 cells, and cell homogenates or membrane preparations were used to determine their glucuronidation ability. UGT2B7(Y) reacted with higher efficiency toward 4-hydroxyestrogenic catechols, whereas UGT1A1 and UGT1A3 showed higher activities toward 2-hydroxyestrogens. UGT2B7(H) catalyzed estrogen catechol glucuronidation with efficiencies similar to UGT2B7(Y). Flunitrazepam (FNZ), a competitive inhibitor of morphine glucuronidation in hepatic microsomes, competitively inhibited catechol estrogen glucuronidation catalyzed by UGT2B7(Y), UGT1A1, and UGT1A3. Buprenorphine, an opioid substrate that reacts at high efficiency with each of these UGTs, was also studied. FNZ competitively inhibited buprenorphine glucuronidation with UGT1A1 and UGT2B7 but had no inhibitory activity toward UGT1A3. This suggests that buprenorphine and 2-hydroxycatechol estrogens react with separate active sites of UGT1A3. A catecholamine, norepinephrine, did not inhibit UGT2B7(Y)-, UGT1A1-, and UGT1A3-catalyzed glucuronidation of catechol estrogens. These results also suggest that drug endobiotic interactions are possible in humans and may have implication in carcinogenesis. PMID- 9848111 TI - Lung tumor induction by inhalation exposure to molybdenum trioxide in rats and mice. AB - Inhalation studies of molybdenum trioxide (MoO3) were conducted because of its wide use in industry, human exposure, and lack of data on carcinogenicity. Groups of 50 male and 50 female F344/N rats and B6C3F1 mice were exposed to MoO3 by inhalation at 0, 10, 30, or 100 mg/m3, 6 h/day, 5 days/week, for 2 years. In both rats and mice, survival and mean body weights of exposed groups of males and females were similar to those of their respective controls. There were significant exposure-dependent increases in blood molybdenum concentration in exposed rats and mice. There were no toxicological differences in bone density or curvature between exposed animals and their respective controls. In rats, dose dependent increases in incidence of hyaline degeneration in the nasal olfactory epithelium and squamous metaplasia of the epithelium lining the base of the epiglottis were observed. The incidence of alveolar/bronchiolar adenoma or carcinoma (combined) was marginally increased in males but not in females compared with controls. In mice, the incidences of squamous metaplasia of the epithelium lining the base of the epiglottis, hyperplasia of the laryngeal epithelium, and metaplasia of the alveolar epithelium were significantly increased in all exposed males and females compared with controls. The incidence of alveolar/bronchiolar adenoma or carcinoma (combined) in exposed groups of males and females was significantly greater than that in the control groups. PMID- 9848112 TI - The role of alcohol dehydrogenase in the metabolism of the colon carcinogen methylazoxymethanol. AB - The aim of this study was to determine whether the cytosolic enzyme alcohol dehydrogenase (ADH) activates methylazoxymethanol (MAM) in the mouse colon and whether differential tumor susceptibility in the mouse is dependent, in part, on strain-related differences in MAM metabolism by ADH. Liver and colon cytosols were isolated from 7-week-old male tumor-susceptible (SWR/J) and -resistant (AKR/J) mice. Minimal reduction of NAD+ was found in colon cytosols from AKR/J mice at the highest concentration (2 mM) of MAM tested. In liver cytosols, only SWR was capable of sustaining NAD+ reduction with MAM, although at very low levels. Despite minimal reactivity with MAM, however, mouse cytosols did effectively reduce NAD+ in the presence of the common ADH subrates ethanol and benzyl alcohol. NAD(+)-coupled oxidation of benzyl alcohol was significantly higher (two- to three-fold, p < 0.05) in mouse colon cytosols compared to activity present within corresponding rat tissues. Incubation of colon and liver cytosols with the ADH-3 inhibitor 4-methylpyrazole markedly (95-100% of controls) reduced ethanol oxidation in both strains. However, 4-methylpyrazole was a less effective inhibitor of benzyl alcohol oxidation in AKR/J colons, suggesting a different ADH isoform complement. An opposite inhibition pattern of benzyl alcohol oxidation was seen in the liver, where 4-methylpyrazole produced a greater inhibition in SWR/J mice. These studies suggest that the metabolism of the proximate mutagen MAM occurs by processes in the mouse that are independent of ADH. PMID- 9848113 TI - Differential sensitivity of chromium-mediated DNA interstrand crosslinks and DNA protein crosslinks to disruption by alkali and EDTA. AB - Some compounds of hexavalent chromium are well-established carcinogens. Chromium enters mammalian cells in the hexavalent form and is reduced to chromium (III). Treatment of purified DNA with chromium (III) produces DNA-DNA interstrand crosslinks (DDC) which obstruct the progression of DNA polymerases in vitro. DDC were also detected in chromate-treated cultured normal human lung cells using the renaturing agarose gel electrophoresis (RAGE) assay and correlated with base specific inhibition of DNA replication. Curiously, DDC have gone undetected in studies of cultured cells using the alkaline elution (AE) technique, whereas chromium-mediated DNA-protein crosslinks (DPC) were readily detected by AE. We tested the hypothesis that AE conditions [60 mM tetraethyl ammonium hydroxide (TEA), 20 mM EDTA, pH 12.6, for 16 h at room temperature] dissociate DDC but not DPC using chromium(III)-treated plasmid DNA and the RAGE assay. Dose-dependent chromium-induced DDC were unaffected by TEA (pH 11.8) alone or by more rigorous alkaline denaturation conditions (200 mM NaOH, pH 13.5, for 16 h). DDC were, however, completely disrupted by EDTA (pH 12.6) alone or the combination of TEA and EDTA (pH 12.6). In contrast, DPC remained largely intact under these conditions. Therefore, past AE-based studies which have failed to detect chromium induced DDC do not prove the absence of this lesion. AE may not be suitable for detecting DDC induced by EDTA-chelatable agents such as metals. PMID- 9848114 TI - Systemic effects of pegylated recombinant human megakaryocyte growth and development factor in combination with recombinant murine granulocyte colony stimulating factor in a murine model of myelosuppression. AB - Megakaryocyte growth and development factor (MGDF) stimulates megakaryopoiesis and thrombopoiesis in vivo. Previous studies indicate that administration of pegylated recombinant human (PEG-rHu) MGDF in combination with recombinant murine granulocyte colony-stimulating factor (rMuG-CSF) prevented lethality and reduced hematotoxicity in carboplatin-treated/irradiated mice, a disease-state animal model of radio-chemotherapy. In the current study we have further characterized the effects of PEG-rHuMGDF in combination with rMuG-CSF with respect to clinical chemistry, hematology variables, and histologic evaluations to determine whether any potential toxicological interaction exists both in normal and myelosuppressed mice. Myelosuppression and subsequent thrombocytopenia in mice was induced with a combination of a single intraperitoneal injection of 1.25 mg carboplatin followed 4 h later with sublethal gamma irradiation exposure of 500 rad. Both normal and carboplatin-treated/irradiated mice were administered daily subcutaneous injections of 50 micrograms/kg PEG-rHuMGDF alone and in combination with 10 micrograms/kg rMuG-CSF for 21 consecutive days. Administration of PEG-rHuMGDF alone or in combination with rMuG-CSF to carboplatin-treated/irradiated mice increased survival 70 and 100%, respectively, and accelerated platelet recovery. Microscopic examination of nonhematopoietic organs showed no evidence of any morphological changes in normal and carboplatin-treated/irradiated animals. In hematopoietic organs clinically significantly increased granulopoiesis and megakaryopoiesis, as well as extramedullary granulopoiesis within the mandibular and mesenteric lymph nodes, were present. The erythroid line was unaffected by cytokine treatment. In normal, non-carboplatin-treated/irradiated mice, platelet counts increased 6 and 12-fold above baseline in the groups administered PEG rHuMGDF alone or in combination with rMuG-CSF, respectively. The results of this study provide a basis for coadministration of PEG-rHuMGDF with Filgrastim (rHuG CSF) in the clinical treatment of myelosuppression induced by radiation and chemotherapy. PMID- 9848115 TI - Inhibitory effect of oleanolic acid on 12-O-tetradecanoylphorbol-13-acetate induced gene expression in mouse skin. AB - Oleanolic acid (OA) has been shown to inhibit mouse skin tumor promotion by 12-O tetradecanoylphorbol-13-acetate (TPA). This study was designed to examine the effect of OA on the TPA-induced expression of the ornithine decarboxylase (ODC) gene as well as other genes. OA inhibited the induction of ODC activity and mRNA level produced by TPA in the skin of female CD-1 mice. Preapplication of OA (10 mumol) to the mouse dorsal skin produced an approximately 50% decrease in TPA (8 nmol)-induced epidermal ODC activity, as well as ODC gene expression. These results suggest that OA inhibits TPA-induced ODC mainly at the transcriptional level. In addition to ODC, TPA also stimulated metallothionein (MT) gene expression in mouse skin. A dose of 2.5 mumol of OA diminished the TPA-induced MT mRNA 50%. Treatment with OA (10 mumol) after TPA (8 nmol) application also inhibited ODC and MT gene expression which suggests that OA does not compete with TPA for its receptor. OA pretreatment also prevented c-fos gene expression. All of these findings suggest that OA diminishes some signal transduction pathways of TPA to suppress target gene expression in mouse skin. This study suggests that OA might be a general inhibitor against TPA-stimulated gene expression in mouse skin. PMID- 9848116 TI - Thyroxine replacement attenuates hypothyroxinemia, hearing loss, and motor deficits following developmental exposure to Aroclor 1254 in rats. AB - The nervous system is dependent upon thyroid hormones for normal development, and we previously reported that developmental Aroclor 1254 (A1254) exposure caused hypothyroxinemia, hearing loss and other behavioral changes in rats. (Goldey et al., 1995a; Herr et al., 1996). The hypothesis that A1254-induced hypothyroxinemia may have contributed to the observed functional changes was tested in primiparous Long-Evans rats given daily oral doses of corn oil (control) or 8 mg/kg of Aroclor 1254 from gestation day (GD) 6 through postnatal day (PND) 21. In addition, from PND 4 to PND 21, all pups in one-half of the litters received daily, subcutaneous injections of saline or 100 micrograms/kg thyroxine (T4), to yield four groups of litters: corn oil plus saline (CO-S),. corn oil plus T4 (CO-T4), Aroclor 1254 plus saline (PCB-S), and Aroclor 1254 plus T4 (PCB-T4). We measured thyroid hormone concentrations (T4 and T3) in serum collected from 7-, 14-, and 21-day-old pups. The kinetics of the injected T4 were also monitored in the CO-T4 and PCB-T4 groups on PND 7 and 21 by measuring T4 and T3 at 1, 3, 5, 8, and 24 h after injection. Circulating T4 concentrations were dramatically depleted in the PCB-S group relative to CO-S. The kinetics study indicated that T4 therapy raised circulating T4 concentrations following in the PCB-T4 pups to near CO-S concentrations, but only for approximately 6 h postinjection, and T4 concentrations fell precipitously thereafter to near PCB-S concentrations. In accord with previous studies, PCB-S pups showed early eye opening, an effect which was exacerbated by T4 injection (in both the CO-T4 and the PCB-T4 groups). Motor activity (figure-eight maze) testing also replicated our finding of an age-dependent, transient reduction in motor activity on PND 15 that was significantly attenuated in the PCB-T4 group. Similarly, we again found reduced acoustic startle amplitudes on PND 23 and low-frequency (1 kHz) hearing loss in animals tested as adults (the latter determined by reflex modification audiometry). Importantly, the hearing loss at 1 kHz in PCB-exposed animals was significantly attenuated by T4 replacement therapy. These data suggest the hypothesis that hypothyroxinemia is involved in PCB-induced alterations in motor and auditory function, while other effects (e.g., eye opening) appear to have a different mechanism of action. PMID- 9848117 TI - Four-week exposure to diesel exhaust induces nasal mucosal hyperresponsiveness to histamine in guinea pigs. AB - It has been reported that diesel exhaust (DE) particulates augment increases in nasal congestion and nasal secretion induced by histamine (His). We also showed that short-term (3-h) exposure to DE induces nasal mucosal hyperresponsiveness to His. Therefore, in the present study we investigated that whether 4-week exposure of guinea pigs to diesel exhaust would likewise induce nasal mucosal hyperresponsiveness to His. Sneezing number, nasal secretion from the nostril, and intranasal airway resistance induced by His were measured as indices of sneezing response, rhinorrhea, and nasal congestion, respectively. Guinea pigs of each group were exposed to filtered air, with or without a low or high concentration of DE for 3, 7, or 28 days. Exposure to a low or high concentration of DE itself did not induce sneezing, nasal secretion, or nasal congestion. However, exposure to a high concentration of DE augmented that the number of sneezes induced by His, whereas exposure to a low concentration of DE had no significant effect. Exposure to DE for 7 and days tended to augment an increase in nasal secretion induced by exposure to His aerosol in a DE concentration dependent fashion. The augmentation, however, was not statistically significant. Exposure to high or low DE for 3 or 7 days had no significant effect on the increase in intranasal pressure (INP) induced by a 10-min exposure to His aerosol, but exposure to high DE for 28 days augmented the increase in INP induced by His, significantly. Exposure to low DE for 28 days did not augment the increase in INP immediately after inhalation of His aerosol. These results reveal that 4-week exposure to high DE induces nasal mucosal hyperresponsiveness in guinea pigs. PMID- 9848118 TI - Subchronic toxicity of human immunodeficiency virus and tuberculosis combination therapies in B6C3F1 mice. AB - Combination therapy with anti-HIV drugs and opportunistic infection drugs is a common practice in treatment of AIDS patients. Although toxic effects of most individual therapies are known, the toxic potential of most combination therapies has not been established. To understand the toxic consequences of combination therapies, the commonly used anti-HIV drug 3'-azido-3'-deoxythymidine (AZT) and tuberculosis infection therapies pyrazinamide, isoniazid, and rifampicin were evaluated by 13-week gavage studies in B6C3F1 mice, either alone or AZT in combination with one of the antituberculosis drugs. The doses include AZT 100, 200, and 400; pyrazinamide 1000 and 1500; isoniazid 50, 100, and 150; and rifampicin 100, 200, and 400 mg/kg/day. AZT alone caused hematopoietic toxicity with dose-related bone marrow suppression, macrocytic anemia, and thrombocytosis. Pyrazinamide or isoniazid alone at the doses tested did not cause significant toxicity. Rifampicin alone caused hematopoietic toxicity and possibly mild hepatic toxicity. Pyrazinamide below 10 times the therapeutic dose when given with AZT did not increase the hematological toxicity of AZT. Isoniazid markedly increased the hematological toxicity of AZT and contributed to mortality at 3 to 4 times the therapeutic dose combinations. Administration of rifampicin with AZT at the calculated therapeutic doses resulted in toxicity of far greater magnitude than that caused by AZT or rifampicin alone. Combination treatment with AZT and rifampicin caused severe anemia with mortality at 2 to 4 times the therapeutic dose combinations. However, AZT did not enhance the hepatotoxicity of rifampicin. Increased hematopoietic toxicity of AZT when given in combination with the above antituberculosis drugs may be due to changes in the metabolism of AZT. Results of these studies indicate that toxicological effects of combination therapies could be considerably more severe than and different from the toxicity of individual therapies. PMID- 9848119 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin in pregnant Long Evans rats: disposition to maternal and embryo/fetal tissues. AB - Prenatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) interferes with fetal development at doses lower than those causing overt toxicity in adult animals. In a multigeneration study (Murray et al., 1979), female rats that were administered 0.01 microgram TCDD/kg/day in their diet did not experience reduced fertility; however, reduced fertility was seen in the F1 and F2 generations. Exposure to TCDD during development produces alterations in the reproductive system of the developing pups, such as delayed puberty and reduced sperm counts in males (Mably et al., 1992a; Gray et al., 1995) and malformations in the external genitalia of females (Gray and Ostby, 1995). Therefore, the objectives of this study were to determine maternal and fetal tissue concentrations of TCDD that are associated with the adverse reproductive effects seen by Gray and co workers. Pregnant Long Evans rats received a single oral dose of 1.15 micrograms [3H]TCDD/kg on Gestation Day (GD) 8 and maternal as well as fetal tissue concentrations of TCDD were measured on GD9, GD16, and GD21. On GD9, the highest level of TCDD localized in the maternal liver (25.1% dose). In addition, the amount reaching all the embryos on GD9 was 0.01% of the administered dose, which resulted in a concentration of 0.02% dose/g. The amount of TCDD reaching the fetal compartment (fetuses + placentas) increased to 0.12% dose/tissue on GD16 and 0.71% by GD21. The concentration of TCDD within the fetal compartment (0.01% dose/g) on GD16 was comparable to that found in the maternal blood and spleen. Concentrations of TCDD in a single embryo/fetus were 39.6, 18.1, and 22.1 pg/g on GD9, GD16, and GD21, respectively. Estimates of hepatic half-life of elimination in pregnant rats suggested that TCDD may be eliminated faster in pregnant LE rats. Therefore, measurements of biliary elimination were made in pregnant and nonpregnant LE rats to compare rates of metabolism; however, biliary elimination of TCDD is not affected by pregnancy. In conclusion, this dose administered during a critical period of organogenesis causes adverse effects on the developing reproductive system of rodents. This dose produced a body burden of 22.1 pg TCDD/g within a single fetus on GD21. This indicates that low-level TCDD exposure during the perinatal stage of life can produce adverse effects within the developing pups. PMID- 9848120 TI - Short-term biomarkers of tumor promotion in mouse skin treated with petroleum middle distillates. AB - Topical application of certain petroleum middle distillates (PMD) to mice produces skin tumors after long latency, and initiation/promotion protocols indicate that this effect is associated with their tumor promoting activity. Since induction of sustained, potentiated epidermal hyperplasia is predictive of promoting activity, five compositionally distinct PMD [hydrodesulfurized kerosene (API 81-07); hydrodesulfurized PMD (API 81-10); odorless light petroleum hydrocarbons; severely hydrotreated light vacuum distillate (LVD); and lightly refined paraffinic oil (LRPO)] were assessed for their effects on epidermal hyperplasia. PMD were administered (2 x/week for 2 weeks) to skin of CD-1 mice. Four quantitative biomarkers of epidermal hyperplasia were evaluated: epidermal thickness, number of nucleated epidermal cells per unit length of basement membrane, labeling (BrdUrd) index of epidermal cells, and induction of epidermal ornithine decarboxylase (ODC) activity. As positive controls, 12-O tetradecanoylphorbol-13-acetate (TPA) and n-dodecane were utilized. PMD-induced skin irritation was evaluated visually and/or histopathologically. All five PMD produced dose-dependent, skin irritation and epidermal hyperplasia. On a weight basis the magnitude of the maximal PMD-induced effects was similar to that produced by n-dodecane, but > 1000-fold less than that produced by TPA. Epidermal hyperplasia and subacute skin irritancy produced by the five PMD were similar. Of the four short-term markers of tumor promotion assessed, labeling index and epidermal ODC activity were predictive of the relative promoting activities of those PMD for which tumorigenicity bioassay data are available, i.e., API 81-07 > API 81-10 > LRPO. An apparent discrepancy to the predictability of epidermal ODC activity occurred with LRPO:toluene [1:1 (v/v)]. This mixture is nontumorigenic, yet significantly induced epidermal ODC activity. This mixture, however, produced severe epidermal toxicity that precluded any meaningful analysis of short-term biomarkers in relationship to biological activity. PMID- 9848121 TI - Oncogenic studies with felbamate (2-phenyl-1,3-propanediol dicarbamate). AB - Felbamate, 2-phenyl-1,3-propanediol dicarbamate, is a novel anticonvulsant that is effective against both chemically and electrically induced seizures in laboratory animals. Oncogenic studies were conducted in mice and rats to establish a preclinical safety profile for this drug. There was an increased incidence of hepatic cell adenoma in male and female mice and in female rats. There was an increased incidence of interstitial cell tumors of the testes in the male rat. PMID- 9848122 TI - Nonlinearities in 2-acetylaminofluorene exposure responses for genotoxic and epigenetic effects leading to initiation of carcinogenesis in rat liver. AB - The dose responses for several effects of low-level limited exposures to 2 acetylaminofluorene (AAF) in the livers of male Fischer 344 rats were measured and a subsequent phenobarbital tumor promotion regimen was used to manifest initiation of carcinogenesis. Three doses over a 10-fold range yielding cumulative total exposures of 0.126, 0.42, and 1.26 mmol AAF/kg body weight were achieved by daily intragastric instillation for up to 12 weeks with interim terminations. This was followed by 24 weeks administration of 500 ppm phenobarbital (PB) in the diet to promote liver tumor development. At 12 weeks at the end of AAF administration, all exposures produced adducts in liver DNA, measured by 32P postlabeling, and the level of adducts increased with exposure, except that the high exposure did not produce a dose proportional increase. Measurement of arylsulfotransferase activity, a key enzyme in the metabolic activation of AAF, revealed that in livers from the high exposure animals, the enzyme was inhibited. To assess for toxicity, the centrilobular zone of glutamine synthetase-positive hepatocytes was quantified immunohistochemically at 12 weeks. The area of the zone was reduced in the high exposure group and there was a trend to reduction in relationship to exposure. The two lower exposures to AAF produced no increase in cell proliferation, whereas the high exposure resulted in a marked increase, about 8-fold over controls. Initiation was assessed by induction of hepatocellular altered foci (HAF) that expressed the placental form of glutathione S-transferase. AAF induced HAF in the high exposure group, 9-fold at 8 weeks and 170-fold at 12 weeks compared to controls. In rats maintained on PB for 24 weeks after exposure, the multiplicity of HAF increased in controls and comparably in the low and mid exposure groups, but remained at the about the same high level in the high exposure group. The high exposure produced a substantial incidence of benign neoplasms by 12 weeks, and with promotion by 36 weeks, all rats developed hepatocellular neoplasia. In the mid exposure group, only one adenoma occurred at 36 weeks in 17 rats, while in the low exposure group, no liver tumor occurred in 23 rats. Thus, these findings document nonlinearities for some of the effects of AAF, with supralinear effects at the high exposure for cell proliferation and induction of HAF, and a no-observed-effect level for induction of promotable liver neoplasms at the lowest cumulative exposure of 0.126 mmol/kg, in spite of the formation of DNA adducts. We conclude that the effects of this DNA-reactive hepatocarcinogen leading to initiation exhibit nonlinearities and possible thresholds. PMID- 9848123 TI - Impaired male sexual development in perinatal Sprague-Dawley and Long-Evans hooded rats exposed in utero and lactationally to p,p'-DDE. AB - Although the pesticide DDT has been banned in the United States for decades, it remains at low levels in the environment. p,p'-DDE, a metabolite of DDT, was recently shown to inhibit the binding of androgens to the androgen receptor and to exert antiandrogenic effects in perinatal Long-Evans (LE) rats at a dose of 100 mg/kg/day administered to pregnant dams. In this study, we compared the effects of p,p'-DDE on male sexual development in offspring of Sprague-Dawley (SD) and LE rats. The chemical was dosed by gavage to pregnant dams at 10 or 100 mg/kg body wt from gestation day 14 to 18. The developing male rats were examined for sexual developmental landmarks, while the effects of p,p'-DDE on androgen receptor expression were evaluated in the testis and other reproductive organs. The tissue dosimetry of p,p'-DDE was also determined at different stages of development following in utero and lactational exposures. The higher p,p'-DDE dose induced a reduction in the male anogenital distance, an increase in retention of male thoracic nipples and alterations in expression of the androgen receptor in either one or both strains. A much weaker response was seen in the lower dose groups. Tissue and body fluid concentrations of p,p'-DDE were similar in the two strains in some tissues but dissimilar in others, particularly in the serum levels. Higher serum p,p'-DDE levels in the LE strain during pregnancy corresponded with an overall greater sensitivity of the LE strain to the antiandrogenic effects of p,p'-DDE. These results support the previous findings of p,p'-DDE antiandrogenicity in LE rats, extend the findings to SD rats, and suggest that the developmental effects of p,p'-DDE on male rat sexual differentiation are minimal at maternal doses below 10 mg/kg/day. PMID- 9848125 TI - In vitro inhibition of thyroid hormone sulfation by polychlorobiphenylols: isozyme specificity and inhibition kinetics. AB - It was recently demonstrated by our laboratory that hydroxylated metabolites of polychlorinated biphenyls (PCB-OHs) are inhibitors of thyroid hormone sulfation. In this study, a more detailed investigation on sulfotransferase isozyme specificity and the kinetics of inhibition was performed. Thyroid hormone sulfation was determined using 3,3'-diiodothyronine (T2) as a substrate, and various sources of sulfotransferase (SULT) enzyme were used; e.g., female and male rat liver cytosol, male brain cytosol and cytosolic preparations of V79 cells transfected with rat SULT1C1, and human SULT1A1 and human SULT1A3. The inhibition pattern and IC50 values were very similar for male and female rat liver and rSULT1C1 and hSULT1A1. PCB-OHs were not able to inhibit the T2 sulfotransferase activity using hSULT1A3. Metabolite 3-hydroxy-2,3',4,4',5 pentachlorobiphenyl did not inhibit T2 sulfotransferase activity in male brain cytosol, while it was a very potent inhibitor in male and female rat liver cytosol. IC50 values for the tested PCB-OHs were not different with either T2 or 3,3',5-triiodothyronine (T3) as substrate, supporting the hypothesis that T2 is the preferred iodothyronine substrate for the sulfotransferases catalyzing the sulfation of the active hormone T3. The Lineweaver-Burk plot obtained with rat liver cytosol and T2 suggested that the nature of the T2 sulfation inhibition by 4-hydroxy-2',3,3',4',5-pentachlorobiphenyl is competitive. Finally, it was demonstrated that tested hydroxylated polychlorinated dibenzo-p-dioxins and biphenyls were, albeit poorly, sulfated by sulfotransferases as measured by the production of 35S-labeled metabolites. PMID- 9848124 TI - A model for pharmacokinetics and physiological feedback among hormones of the testicular-pituitary axis in adult male rats: a framework for evaluating effects of endocrine active compounds. AB - The testicular-hypothalamic-pituitary axis controls reproductive functions in males. A description of the basic physiological interactions in adult rats among testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) was developed, permitting simulation of hormone levels in testes and blood. This model was used to simulate hormone levels in intact, castrate, ethane dimethanesulfonate-treated, and antiandrogen-treated rats. A large gradient of testosterone concentrations from testicular interstitial fluid to low levels in peripheral blood is created by the testicular blood flow. The dominant feedback loop is positive regulation of testosterone synthesis by LH and negative feedback of testosterone on LH and FSH. The utility of the model for placing in vitro data in the context of in vivo physiology was illustrated for the case of continued synthesis of testosterone by the isolated testes. In the absence of blood flow, very low residual testosterone synthesis can substantially increase testosterone concentration in isolated testes. Effects of an exogenous endocrine active compound were illustrated by modeling altered LH and FSH regulation by testosterone in the presence of an antiandrogen acting as a competitive ligand for the androgen receptor. Increasing concentrations have no effect on steady state hormone levels until sufficient levels of antiandrogen are achieved to reduce negative feedback of testosterone on LH and FSH. In summary, a model has been developed that provides a basis for initiating evaluations of key issues of concern for the risk assessment of endocrine active compounds including in vitro to in vivo extrapolation and their dose-response behaviors. PMID- 9848126 TI - Allergic responses to the biopesticide Metarhizium anisopliae in Balb/c mice. AB - Metarhizium anisopliae is used as a microbial pesticide to control cockroaches and other insects. M. anisopliae has demonstrated neither infectivity nor toxicity in mammals. However, allergenicity has not been assessed. M. anisopliae is a prototype for other organisms released into the environment for pesticide or other beneficial applications. Hence this study is part of an effort to develop methods for screening such organisms for allergenic potential. Soluble factors from fungal components were combined in equal protein amounts to form a crude fungal antigen (MACA). Balb/c mice were intratracheally (IT) challenged with 25 micrograms fungal antigen 13 days post intraperitoneal sensitization with the fungal antigen in alhydrogel adjuvant. Additionally, mice were sensitized with adjuvant alone or chitin media in adjuvant as experimental controls. Serum and bronchoalveolar lavage fluid (BALF) were harvested prior to challenge and at 1 and 7 days post IT challenge (DPIT). These mice exhibited immune and pulmonary inflammatory responses to MACA characteristic of allergy. Total serum IgE for antigen-sensitized animals increased 7.6- and 14.7-fold over that for chitin media and adjuvant controls, respectively, at 7 DPIT. Less striking increases were seen at 24 DPIT and prior to challenge. BALF IL-4 was dramatically elevated only in MACA-sensitized and challenged mice and only at 1 DPIT. Additionally, there was a dose-dependent increase in BALF eosinophils from MACA-sensitized mice at both 1 and 7 DPIT. While lymphocyte counts were increased for all treatment groups at 1 DPIT, by 7 DPIT lymphocyte counts for MACA-sensitized mice only were significantly elevated compared to controls. Pulmonary inflammation, edema, and cell damage were apparent at 1 DPIT (25 micrograms MACA), as indicated by a neutrophilic influx and elevated levels of total protein and LDH, in both sensitized and control groups. These effects were significantly decreased, but not eliminated by reduction of the challenge dose to either 10 or 5 micrograms MACA. While BALF IL-4 was also reduced at the lower challenge doses, eosinophilia and total IgE were unchanged. The data suggest that the crude fungal extract MACA contains one or more potent allergens and that total IgE may be useful in the identification of the allergen(s). PMID- 9848127 TI - Effect of glutathione depletion and metallothionein gene expression on arsenic induced cytotoxicity and c-myc expression in vitro. AB - Arsenic exposure is clearly linked to human cancer. In rodent cells, arsenic has been reported to induce aberrant gene expression, including activation of the proto-oncogene c-myc. Abnormal or altered expression of such oncogenes can be involved in the acquisition of a malignant phenotype. Although its mechanism of action is unclear, arsenic is known to exert at least some of its toxic effects through interaction with sulfhydryl groups, and the non-protein sulfhydryl glutathione (GSH) appears to play an important role in detoxication of arsenic. Similarly, metallothionein (MT), a metal-binding protein with high sulfhydryl content, often functions in defense against metal-induced or oxidative cellular injury. Therefore, we examined the relationship among GSH, MT gene expression, and arsenic-induced toxicity or c-myc expression in cultured rat myoblast (L6) cells. In initial toxicity studies, arsenic was used in both the trivalent (arsenite) and pentavalent (arsenate) forms. The role of GSH was studied by pretreating cells with L-buthionine sulfoximine (BSO), which induces a marked depletion of GSH. In vitro exposure of L6 cells to BSO (1 to 25 microM) resulted in dose-dependent decreases in GSH. GSH depletion sensitized cells to both arsenite and arsenate. Zinc pretreatment, at levels which highly activated MT expression, had no effect on arsenite-induced cytotoxicity. Arsenite (1 microM) alone modestly increased c-myc expression from 1 to 4 h after treatment (maximum of 2.0-fold over control). After GSH depletion cells responded to arsenite exposure with much larger increases in c-myc transcription (3.2-fold over control). Zinc pretreatment had no reductive effect on arsenite-induced c-myc expression despite markedly activating the MT gene. Thus, it appears that the cellular levels of GSH, but not MT gene expression, play an important role in resistance to arsenic toxicity and aberrant gene activation. Moreover, depletion of GSH enhances arsenic-induced proto-oncogene activation, which might contribute to subsequent transformation. PMID- 9848128 TI - Assessment of the developmental toxicity, metabolism, and placental transfer of Di-n-butyl phthalate administered to pregnant rats. AB - The developmental toxicity and placental transfer of di-n-butyl phthalate (DBP) were evaluated in Sprague-Dawley rats given a single oral dose of DBP on Gestational Day 14. In the developmental toxicity study, dams were dosed with 0, 0.5, 1, 1.5, or 2 g DBP/kg and were necropsied on GD21. Increased incidence of resorptions and reduced fetal body weight were observed at 1.5 and 2 g/kg. Higher incidences of skeletal variations were found at doses > or = at 1 g/kg. No embryotoxic or teratogenic effects were observed at a dose of 0.5 g/kg. In the placental transfer study, dams were dosed with 0.5 or 1.5 g [14C]DBP/kg. Maternal and embryonic tissues were collected at intervals from 0.5 to 48 h. Embryonic tissues accounted for less than 0.12-0.15% of the administered dose. Levels of radiocarbon in placenta and embryo were one-third or less of those in maternal plasma. No accumulation of radioactivity was observed in the maternal or embryonic tissues. From HPLC analyses, it was shown that unchanged DBP and its metabolites mono-n-butyl phthalate (MBP) and MBP glucuronide were rapidly transferred to the embryonic tissues, where their levels were constantly lower than those in maternal plasma. MBP accounted for most of the radioactivity recovered in maternal plasma, placenta, and embryo. Unchanged DBP was found only in small amounts. These findings support the hypothesis that MBP, a potent teratogen, largely contributes to the embryotoxic effects of DBP. PMID- 9848129 TI - Acute, subchronic, and chronic toxicity studies with felbamate, 2-phenyl-1,3 propanediol dicarbamate. AB - Felbamate, 2-phenyl-1,3-propanediol dicarbamate, is a novel anticonvulsant that is effective against both chemically and electrically induced seizures in laboratory animals. Acute, subchronic, and chronic studies were conducted in mice, rats, and dogs to establish a preclinical safety profile for this drug. Clinical signs following single intraperitoneal doses included hypoactivity, tremors, decreased muscle tone, ataxia, prostration, and labored breathing. Death was observed after intraperitoneal but not oral administration. A consistent drug related effect noted in all multiple-dose studies with this compound was decreased body weight and food consumption. The only other consistent change noted in multiple-dose studies with felbamate was an increase in liver weight (relative and absolute) in the rat and dog which was accompanied in some cases by increases in serum enzyme levels. No histopathological changes were observed in the liver that could explain these elevated serum enzyme levels. Based on the results of these studies it was concluded that long-term administration of felbamate in human clinical trials was warranted. PMID- 9848130 TI - Temporal changes in tissue repair permit survival of diet-restricted rats from an acute lethal dose of thioacetamide. AB - Although, diet restriction (DR) has been shown to substantially increase longevity while reducing or delaying the onset of age-related diseases, little is known about the mechanisms underlying the beneficial effects of DR on acute toxic outcomes. An earlier study (S. K. Ramaiah et al., 1998, Toxicol. Appl. Pharmacol. 150, 12-21) revealed that a 35% DR compared to ad libitum (AL) feeding leads to a substantial increase in liver injury of thioacetamide (TA) at a low dose (50 mg/kg, i.p.). Higher liver injury was accompanied by enhanced survival. A prompt and enhanced tissue repair response in DR rats at the low dose (sixfold higher liver injury) occurred, whereas at equitoxic doses (50 mg/kg in DR and 600 mg/kg in AL rats) tissue repair in AL rats was substantially diminished and delayed. The extent of liver injury did not appear to be closely related to the extent of stimulated tissue repair response. The purpose of the present study was to investigate the time course (0-120 h) of liver injury and liver tissue repair at the high dose (600 mg TA/kg, i.p., lethal in AL rats) in AL and DR rats. Male Sprague-Dawley rats (225-275 g) were 35% diet restricted compared to their AL cohorts for 21 days and on day 22 they received a single dose of TA (600 mg/kg, i.p.). Liver injury was assessed by plasma ALT and by histopathological examination of liver sections. Tissue repair was assessed by [3H]thymidine incorporation into hepatonuclear DNA and proliferating cell nuclear antigen (PCNA) immunohistochemistry during 0-120 h after TA injection. In AL-fed rats hepatic necrosis was evident at 12 h, peaked at 60 h, and persisted thereafter until mortality (3 to 6 days). Peak liver injury was approximately twofold higher in DR rats compared to that seen in AL rats. Hepatic necrosis was evident at 36 h, peaked at 48 h, persisted until 96 h, and returned to normal by 120 h. Light microscopy of liver sections revealed progression of hepatic injury in AL rats whereas injury regressed completely leading to recovery of DR rats by 120 h. Progression of injury led to 90% mortality in AL rats vs 30% mortality in DR group. In the surviving AL rats, S-phase DNA synthesis was evident at 60 h, peaked at 72 h, and declined to base level by 120 h, whereas in DR rats S-phase DNA synthesis was evident at 36 h and was consistently higher until 96 h reaching control levels by 120 h. PCNA studies showed a corresponding increase in cells in S and M phase in the AL and DR groups. DR resulted in abolition of the delay in tissue repair associated with the lethal dose of TA in ad libitum rats. Temporal changes and higher tissue repair response in DR rats (earlier and prolonged) are the conduits that allow a significant number of diet restricted rats to escape lethal consequence. PMID- 9848131 TI - Effects of dexamethasone on functional and pathological changes in rat bronchi caused by high acute exposure to chlorine. AB - We assessed the effects of dexamethasone on functional and histological changes after acute exposure to a high level of chlorine gas in an animal model of reactive airways dysfunction syndrome (RADS). Sprague-Dawley male rats were exposed to 1500 ppm of chlorine for 5 min and treated with either dexamethasone (dex; 300 micrograms/kg/day) or saline intraperitoneally for 7 days. Lung resistance (RL), airway responsiveness to inhaled methacholine (MCh), airway wall morphometric measurements, and bronchoalveolar lavage (BAL) cells were assessed over a 2-week period after exposure. Dex administration significantly attenuated both chlorine-induced increased RL and chlorine-induced increased responsiveness to methacholine compared with saline: -2.7 +/- 6.8% vs 102.3 +/- 36.6% change from baseline RL (P < 0.01) and 2.5 +/- 0.6 mg/ml vs 1.2 +/- 0.7 mg/ml in the MCh concentration required to double the RL from baseline (P < 0.01). There was a tendency, albeit nonsignificant, for improvement in some indices of epithelial injury. Dex significantly attenuated the postexposure neutrophilic cellular response in BAL 1 day after exposure (15.8 +/- 4.9% neutrophils in the dex group vs 49.8 +/- 2.7% neutrophils in the saline group) (P < or = 0.001). Our results show that dex administration helps maintain pulmonary function, reduces BAL inflammatory cell number, and tends to improve some morphometric airway wall structure parameters in rats exposed to chlorine. PMID- 9848132 TI - The heart rate-corrected QT interval of conscious beagle dogs: a formula based on analysis of covariance. AB - Three frequently used and cited formulas used to rate correct the QT interval (Bazett's, Fridericia's, and Van de Water's) were compared and ranked using a large population-based cohort of beagle dogs (99 males and 99 females). In addition, analysis of covariance was used to derive a flexible method to rate correct the QT.interval for heart rate. The method is flexible in that it utilizes pretest or control data to determine the degree of correction. In addition, it can also be used to evaluate whether treatment alters the association between heart rate and QT. Specifically, pretest QT (unadjusted) and heart rate data were used to estimate coefficients in the linear regression log(QT) = alpha + beta log(HR). The estimated slope (beta) from the pretest data was used to heart rate correct the QT interval in the formula log(QT)ca = log(QT) - beta *[log(HR - log(HRm)]. The term "log(HRm)" is included to standardize QTca to a reference value, either a fixed value or an average heart rate for the data set being analyzed. These formulas were retrospectively compared under a typical toxicity study paradigm with a class III antiarrhythmic agent (L-768,673) that selectively prolongs the QT interval by blocking the slow activating component of the delayed rectifying potassium channel (lks). Based on their ability to dissociate the effects of heart rate on the QT interval, the formulas received the following ranking: Covariate Adjustment (preferred) = Van De Water's > Fridericia's > Bazett's (not recommended). Analysis of covariance based on pretest or control data is preferred for moderate to large studies where there are adequate data for estimation of the slope parameter beta, the investigator does not have sufficient control over HR, or treatment alters the association between HR and the QT interval. Conversely, for smaller studies a fixed rate adjustment formula from the literature (such as Van de Water's or Fridericia's equations) may be preferable since the bias from using a fixed formula is likely to be smaller than the variance resulting from estimating beta from a small sample. PMID- 9848134 TI - [Key enzymes of biosynthesis of amino acids of the glutamic series in the virus cell system Anabaena variabilis + A-1]. AB - The influence of cyanophage A-1 reproduction on glutamate dehydrogenase (GDG) and glutamine synthetase (GS) in A. variabilis cells was studied. It was determined that the both enzymes are intensified by 70% and 30%, accordingly. Isoenzymes of GDG and GS were isolated from native and infected cells of cyanobacteria, they had various physicochemical properties. It is concluded that cyanophage development causes the specific modification of cell enzymes. PMID- 9848133 TI - [Interaction of adhesion proteins with structural components of cytoskeleton in animal cells]. AB - The short characteristic of cell adhesion molecules and the main components of cytoskeleton are submitted at the present review. Structural and functional peculiarities of these proteins are presented. On the basis of different sources an attempt to clarify the possible mechanism of the interaction of adhesion proteins cytoskeleton structure has been done. It is suggested that information about adhesion contacts is transmitted from the adhesion proteins to cytoskeleton structures that induces the transformation of the letter ones and promotes the cellular metabolic response. PMID- 9848135 TI - [Cytosolic thiamine triphosphatase from bovine brain. 2. Interaction of thiamine triphosphate ester with the enzyme]. AB - The analysis of the steady-state kinetics of the thiamine triphosphate ester hydrolysis reaction catalyzed by homogeneous thiamine triphosphatase (EC 3.6.1.28; thiamine triphosphate phosphohydrolase) from bovine brain enables us to suggest, that the ThTP binding to the catalytic site of the ThTPase active centre takes place by the phosphate radical. The correct orientation of the substrate molecule occurs by means of the contact of the thiamine component. The crucial role in this process belong to the amino group of the pyrimidine ring and hydrophobic forces. The quaternary nitrogen of thiazole is important for the hydrolytic splitting of the substrate. The hydrolysis of thiamine triphosphate ester occurs through the formation of the ternary enzyme-substrate complex, with the Mg2+ and Mg.ThTP adding being random. PMID- 9848136 TI - [Trypsin inhibitors of wheat seedlings infected and treated with salicylic acid]. AB - Using the method of affinity chromatography for trypsin-sepharose, followed by PAGE electrophoresis the component composition of trypsin inhibitors was studied. The inhibitors were extracted from the winter wheat seedlings which were infected with Fusarium fungus and also from the seedlings treated with salicylic acid. Under the action of pathogen not only a quantitative increase in the protein components of trypsin inhibitor but also the appearance of a new component with a molecular weight of 29 kDa are observed. The influence of Fusarium graminearum and salicylic acid on the amino acid composition of trypsin inhibitors has been studied. After the inhibitors purification their antienzyme spectrum was studied along with their influence on the pathogen proteases. It was established that the inhibitors influence only the activity of trypsin and chymotrypsin and do not influence the activity of pepsin, elastases, papain, proteinases from the wheat seedlings. The existence of arginine and absence of lysine was demonstrated in the active centre of the inhibitors. PMID- 9848137 TI - [Effect of ammonium chloride and urea on the activity of lactate dehydrogenase. A general model of catalysis]. AB - The dependence of structural and functional properties of lactate dehydrogenase from the pig muscle (isoenzyme M4) on pH (6.9-8.5) on the effect of different forms of non-protein nitrogen (e.g. urea and ammonium chloride) has been investigated. It was shown that Vmax changes negligibly at pH 6.9-8.5 but K(m) increase markedly with pH elevation. At the alkali value of pH pyruvate inhibition disappeared. We have found that urea and ammonium chloride at the non saturation concentrations inhibited the activity of lactate dehydrogenase. The inhibition constants (Ki and Ka) for urea and ammonium chloride were obtained from inhibition data. When analysing works of different authors and own results we propose the general model of catalysis of lactate dehydrogenase, which includes the formation of normal enzyme-substrate complex, abortive enzyme-NAD pyruvate complex and the effect of different effectors, including nitrogen containing substances. PMID- 9848138 TI - [Comparative study of physical properties of apical and basolateral membranes of small intestine enterocytes affected by ionizing radiation]. AB - The physical properties of apical and basolateral membranes of rats small intestine enterocytes after X-ray irradiation in doses of 1.0 and 2.0 Gy were studied. More pronounced decrease of the boundary lipids fluidity, the reduction of the protein component, participating in energy transfer, and immersion of such proteins into lipid phase was revealed in apical membrane in comparison with basolateral membrane. PMID- 9848139 TI - [Effect of estradiol on phosphoinositide composition of nuclear membranes of rat brain cells]. AB - The effect of estradiol on phospholipids and particularly on phosphoinositides of nuclear membranes of rat brain cells at different stages of hormone action was investigated. It was shown that hormone leads to the elevation of phospholipids total amount at the primary and early stages of its action (1 and 4 h after estradiol injection, correspondingly). The increase of content of almost all individual fractions of phospholipids as well as phosphatidylinositol (i.e. monophosphoinositide) was detected. It is remarkable that the monophosphoinositides elevation takes place mainly due to the decrease of di- and especially of triphosphoinositide. The ratio of triphosphoinositide/monophosphoinositide decreases more than twice. It was concluded that the ratio of triphosphoinositide to monophosphoinositide may be a peculiar indicator of the status of nuclear membranes for the functioning of triphosphoinositide regulatory system. PMID- 9848140 TI - [Ca2+/H+ exchange in the lymphocyte plasma membrane]. AB - We studied delta pH-induced Ca(2+)-transport through the lymphocytes plasma membrane. The simultaneous and reversal H(+)-counter transport of Ca2+ was shown in the presence of known Ca(2+)- and H(+)-transporting pathways through the plasma membrane. Ca2+/H+ exchange through the plasma membrane was shown experimentally on the plasma membrane of lymphocytes, obtained from fresh rats' blood under the conditions of inhibition for known Ca2+ and H+ conductive pathways. At least one of the transporting cations' concentration gradient through the outer plasma membrane was only the moving force for the such exchange. Cytoplasmic H(+)-buffering capacity changing and associated plasma membrane's delta pH changing are discussed to imply on redistribution of Ca2+ through the plasma membrane. We suggest the presence of Ca2+/H+ antiporter in the lymphocytes' plasma membrane. PMID- 9848141 TI - [DNA synthesis in lymphocytes of irradiated animals as affected by mitogens and Staphylococcal antigens]. AB - Proliferation activity of lymphocytes from rats was studied using micromethod of blast transformation reaction after exposure of X-ray irradiation (absorbed doses for animals are 0.5 and 1 Gr). The reducing of the functional activity of lymphocytes was found. There was difference in lymphocyte response induced by standard mitogens (PHA, ConA, LPS) and antigens of Staphylococcus protein A and peptidoglycan. The irradiation protective effects of Staphylococcus substances were found. PMID- 9848142 TI - [Protective effect of cresacin in D-galactosamine-induced acute experimental hepatitis]. AB - Antioxidant properties of cresacin were studied on the model of galactosamine hepatitis and on the isolated liver cells of white male rats. It has been shown, that cresacin in a dose of 20 mg/kg effectively inhibits the processes of lipid peroxidation induced by hepatotoxin. Cresacin also normalized some components of fermentative and non-enzymatic antioxidant system. In particular the indexes of the activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and the level of reduced glutathione, total phospholipids and ascorbic acid was increased for certain. In vitro, on the isolated hepatocytes, cresacin showed the dose-dependent antioxidant effect. This fact is confirmed by its property to inhibit the speed of formation of the malonic dialdehyde in the incubation medium. PMID- 9848143 TI - [Biotransformation of lignocellulse by the fungi Pleurotus floridae (Fries) Kummer and Phellinus igniarius (Linnearus:Fries) Quelet--the pathogens of white rot in trees]. AB - The peculiarities of lignocellulose biotransformation by white-rot fungi Pleurotus floridae and Phellinus igniarius during their solid-state fermentation of wastes of oil-bearing crops processing has been studied. The dynamics of oil bearing crops processing wastes bioconversion has been studied. It has been marked that P. floridae utilized 20% cellulose and lignin during 9 weeks and 40% lignin and 30% cellulose during all period of fermentation (19 weeks). The fungus P. igniarius utilized mainly cellulose (40% cellulose and 24% lignin in 19 weeks). Lignin-degradative capacity of P. floridae (KL = 0.57) and P. igniarius (KL = 0.34) has been quantitatively estimated. The degradation of plant biopolimers corresponded to the production of ligninolytic (laccase, Mn(2+) dependent peroxidase,) and cellulolytic (CMC-cellulase) enzymes. The component and isoenzyme content of ligninolytic enzyme complex of fungi Pleurotus floridae and Phellinus igniarius has been determined. PMID- 9848144 TI - [Lipids and their antioxidant activity in tissues of ruminant animals]. AB - The work deals with the estimation of lipids in the bovine tissues (liver, intestine, skin) of the animals bred for the industrial purposes. The tests displayed, that depending on ration, breeding conditions and providing the animals organism with vitamins, the lipids levels in the liver, intestine and skin changes. As a result of the experiments the conclusion was made that deficiency of fatty-soluble vitamins A, D, E in the animals fodder provided for the disturbance of lipid metabolism in their organism and for the development of a pathologic process: a light form of alimentary parathyroid-osteodistrophia. This state was eliminated by means of additive intramuscular injection of the above listed vitamins. Estimation of the test animals tissue lipids properties revealed, that the provision of the organism with the vitamins led to the increase of the level of some natural antioxidants. Normal development of animals is provided when they are fed by natural products. PMID- 9848145 TI - [Antioxidants of blood enzymes in kidney function disorders in man]. AB - The activities of lipid peroxidation processes and enzymes of antioxidant system in the blood of patient with renal failure have been studied. The results demonstrated stimulation of lipid peroxidation processes in blood serum, cellular membranes of erythrocytes and in urine. The study of the antioxidant system discovered the inhibition of various enzymes: superoxide dismutase and peroxidase, inhibiting production of active oxygen forms; transferring and ceruloplasmin as regulators of Fe2+/Fe3+ ratio, glutathione reductase and glutathione peroxidase influencing the ratio of oxidated and reduced glutathione forms and the level of SH-groups. So, determination of lipid peroxidation processes and antioxidant enzymes in blood is the sensitive inform test in diagnostics of chronic renal failure. PMID- 9848146 TI - [Effect of carbon dioxide on free radical processes as affected by artificial hypobiosis in rats]. AB - High CO2 concentrations in the inspired air have been studied for their effect on the free-radical oxidation of lipids in the lipids in the blood, its serum and mitochondria of the rat liver under artificial hypobiosis. Kinetics of free radical oxidation of lipids was estimated from the changes in chemiluminescence parameters as well as the content of products which react with thiobarbituric acid. It is found that in animals in the state of artificial hypobiosis the intensity of free-radical oxidation of lipids was rather less as compared with animals which did not fall into hypobiosis because of the exclusion of CO2 from the inspired air. The data obtained permit concluding that the increase of CO2 concentration in the inspired air prevents from the development of the oxidative stress. It is evident that the increase of CO2 in the inspired air occurs before the formation of the adaptational mechanisms, directed to the preventing of lesion of membrane lipids under artificial hypobiosis. PMID- 9848147 TI - [Immunoglobulins of cows of different ages and of their calves]. AB - The level of IgG1 in blood in cows of different ages before calving is decreased. At antigens stimulation the content of the IgG1 increases in about twice at the expense of specific antibodies connected with antigens which are and transferred into colostrum. The maximum antibodies (IgG1) level is marked in colostrum of mature cows (2-7 lactations) and it reaches of 82.8 mg/ml and the minimum level in the first-born calves-51.6 mg/ml. Correspondingly the calves from the first born cows assimilated 28-36% less immunoglobulins than the calves of the same ages from the cows of older ages. PMID- 9848148 TI - [Pectin production from burdock (Arctium sp.)]. AB - The pectin complex of different morphological parts of burdock: leaf, leaf stems, main stock have been investigated. High quantity of pectic substances about 1.9% per wet mass, has been detected in the leaf stems of burdock. It has been also established that pressed leaf stems as to pectin content (about 21% of dry mass) do not give in to traditional raw material for pectin production: pressed apple, squeezed sugar-beet, citrus fruit peel, sunflower head, etc. The technological scheme of pectin production from burdock has been developed. Pectin powder obtained from burdock was of light grey colour. The total output was 17% of dry mass of raw material. Uronide component of burdock pectin was 66%, esterification rate 47%, methoxyl component 9%, equivalent weight 343, the free carboxyl group content 16%. PMID- 9848149 TI - [Effect of sulfur nutrition for sulfoxide accumulation in garlic bulbs]. AB - The influence of the sulphur nutrition conduction upon sulphoxides accumulation in the garlic bulbs was studied. It was found that application of sulphur as ammonium sulphate in quantity up to 75-100 kg S/ha induced proportional increase of sulphoxides in bulbs from 3.03 up to 4.57 mg/g. Subsequent increasing of the sulphur doses applied did not induce any trustworthy changes. When potassium sulphate was used for feeding similar results were obtained. The potassium and nitrogen increase equivalent to the amounts of them applied together with sulphur did not induce any trustworthy changes too. Also it was registered the garlic variety sulphur nutrition specificity, that was closely connected with degree of the floral spear development. So, in the bulbs of non-spear-forming variety (Ukrainian white) the maximal increasing of the sulphoxide content was 78.6% against control whereas for Boguslavsky 10, the variety with most advanced floral spear (with height up to 125 cm), the increasing of sulphoxides was only 43.6%. PMID- 9848150 TI - [Phospholipid composition of different rat brain regions in acute alcoholic intoxication]. AB - The effect of acute ethanol intoxication (25% solution, intragastrically, 4 g/kg, 1 h) on the concentration of phospholipid fractions in different rat brain regions was studied. It was found that acute alcohol intoxication decreased the level of phosphatidylcholine and increased the concentration of phosphatidylethanolamine in the cerebellum of experimental animals. The cortex of large hemispheres showed a decreased amount of phosphatidylcholine and increased one of sphingomyelin. Possible mechanisms of these changes are discussed. PMID- 9848151 TI - [Protective role of the glutathione system in rat organs after administration of embiquine]. AB - Dynamics of changes of the reduced glutathione contents, activity of glutathione transferase, glutathione reductase and gamma-glutamyl transpeptidase in rat organs at a single injection of embiquine in a dose of 1/2 LD50 has been studied. It was shown that injection of the alkylating antitumor preparation lead to the decrease of the reduced glutathione contents in the early period of intoxication together with compensatory increase of tripeptide concentrations 3 hours later in the liver tissue and 24 hours later in the kidney tissue. Embiquine caused the steady increase of the glutathione transferase activity in all the organs studied. Significant increase of liver, kidney and spleen gamma-glutamyl transpeptidase by the end of the investigation period was noted at the injection of the alkylating agent. PMID- 9848152 TI - [Delayed determination of ceruloplasmin activity in donors' blood serum after adding chemical stabilizer]. AB - The ceruloplasmin activity in bioassays of donor blood serum, preserved at the temperature of 4-8 degrees C, decreases progressively, beginning from the third day of experiment. Freezing of bioassays is accompanied by similar changes, since this enzyme belongs to cryounstable proteins. It has been found out that the 1 triphenylphosphonomethylnaphthalene bromide in concentrations 1.03-2.58 mmol/l stabilizes ceruloplasmin and preserves its activity unchanged in biologic fluids during 30 days of observation. Having used this fluid we have created a delayed method of determining this enzyme which may be used with the aim of studying metabolic processes in the human body of spacemen, submarine crews, members of arctic, antarctic and other expeditions which cannot be accompanied by a biochemical laboratory. PMID- 9848153 TI - [Use of a fluorescent method for detection of lymphocyte Fc-receptors]. AB - FcR on chicken blood lymphocytes have been studied by fluorimetric binding assay. Binding data were analyzed by Scatchard's method. Temperature and time variations were investigated in order to optimise the binding conditions. It was concluded that binding FcR to IgG on chicken blood lymphocytes was optimal at 0 degree C for 60 min. 10-14% of nonspecific binding were detected in different experiments. Scatchard binding plots at 0 degree C demonstrated that IgG bound to FcR per cell 2.3 +/- 4 x 10(5) with affinity 3.9 +/- 0.5 x 10(5) M-1. PMID- 9848154 TI - [Spectrophotometric method for determination of sulfoxides]. AB - Easy and exact method for determination of sulfoxides based on the property of this substances to react with the product of tryptophan's and p dimethylaminobenzaldehyde's condensation is proposed. An oxidizer-deoxidizer reaction results in formation of bis-2-tryptophanyl-4-dimethylaminophenylcarbinol blue-violet dye with the absorption maximum at 575 nm. Sensibility of method is 5 mkmol/g, relative mathematics error at three determinations is +/- 3.3%. Method can be used for studying of features of sulfoxides' biosynthesis and accumulation in garlic tissues and for estimation of quality of bulbs as row material for food and pharmaceutical industry. PMID- 9848155 TI - [Forty years of studying angiotensin-converting enzyme: problems and accomplishments]. AB - Questions on the molecular forms multiplicity, certain physical, chemical and catalytical properties, the mechanisms of the activity regulation and biological role of the angiotensin-converting enzyme have been considered. Special attention was paid to the consideration of the angiotensin-converting enzyme role in brain regulative neuropeptide metabolism. PMID- 9848156 TI - [Retinol transport into the cell nucleus in vitro]. AB - The retinol interaction with intact cellular nuclei, nuclear envelope and chromatin was investigated. We have shown that the Cellular Retinol-Binding Protein (CRBP) plays a very important role in such interaction. Retinol can specifically interacts with nuclei, nuclear envelope and chromatin only when it presents as a complex with CRBP. The obtained data allowed us to suggest that the process of retinol delivery to hypothetical nuclear receptors include of at least the following stages: 1) specific binding with nuclear envelope; 2) penetration into nuclei; 3) specific binding with chromatin receptors. CRBP is a necessary component at all this stages. Also we show that CRBP has not a species or tissue specificity. PMID- 9848157 TI - [In vitro study of the early membrane effects of C27-steroid hormone ecdysterone, immobilized on the nanodispersed magnetite surface]. AB - The object of the study was to reveal the existence of receptor interactions of C27-steroid hormone ecdysterone with the surface of various cells (liver and spleen macrophages, thymus and spleen lymphocytes, erythrocytes and hepatocytes from rat organs) in experiments in vitro using ecdysterone immobilized on the nanodispersed iron surface (which did not penetrate the cell during observation) and the model of fast (during 15 s) activation of phospholipid signal system with ecdysterone. The parameters, which are characteristic of cell phospholipid signal system activation, were evaluated (the concentrations of free arachidonic acid, diene conjugates, tromboxane B2 and leukotriene C4) and its alteration in response to free and immobilized ecdysterone introduction in suspensins of intact cells and cells with the surface modified in vivo by pre-injection of antigen (human erythrocytes). Concurrent binding of immbilized ecdysterone with intact rat cells in the presence of free ecdysterone was also investigated. It was established that there are high-affinity ecdysterone-specific binding sites on the plasma membranes of all cells investigated, Kd estimated according to two different methods varied in the region of 10(-8)-10(-7) M. The characteristic features of the changes in parameters characteristic of phospholipid system activation were revealed to be queite similar in intact cells, as well as in cells with modified surfaces, under introduction of free ecdysterone and immobilized ecdysterone, which did not penetrate into cytosole. The data obtained point to the existence of ecdysterone receptors on the surface of cells plasma membranes and are indicative of the membrane effects as mechanisms of the early pregenomic phase of cells activation by ecdysterone. PMID- 9848158 TI - [Effect of bee venom and its components on electrophoretic mobility of erythrocytes]. AB - The change of electrophoretic mobility (EPM) of erythrocytes of donor blood caused by their incubations with the bee venom and its major components--melittin and phospholipase A2 was investigated. It was estimated that the bee venom reversibly decreased EPM of erythrocytes to the higher degree than its separate components. The reaction was completely abolished after preliminary processing of erythrocytes with neuroaminidase and partly after their processing with glutaraldehyde, but the effect of EPM increased under the conditions of the increasing of ATP concentration. Proceeding from the authors' data it was concluded that the major components of the bee venom successively interacted with glicocalyx elements and membrane of erythrocytes which evokes the charge redistribution in these blood cells. PMID- 9848159 TI - [Effect of alpha-tocopherol and its derivatives on the viability of cultured malignant cells]. AB - The effect of alpha-tocopherol and its derivatives on viability and growth of attached cells culture: human epithelioid carcinoma of cervix HeLa-S3K and human cervical carcinoma cell line C-4-1 according to protein content and level of alkaline phosphatase, as well to unattached cells: human acute leukemia cell cultures CEM-C-1 and CEM-C-7 according to the amount of viable cells in suspension has been studied. It has been shown that alpha-tocopherol acetate and tocopherol maintain the viability of cells, while the derivatives of tocopherol with shorted side chain considerably inhibit growth of researched cultures, alpha tocopherol with lacking side chain being most active among them. PMID- 9848160 TI - [Effect of alpha-tocopherol on the respiratory burst of neutrophils, blast transformation of lymphocytes, and activity of human natural killer cells in blood]. AB - The effect of vitamin E on immunological reactions of blood cells was studied. The addition of vitamin E in concentration of 100 microM to neutrophils caused the increase of superoxide production. But this index was decreased when incubation of these cells with A23187 or FMLP was accompanied by alpha-tocopherol in concentration of 100 microM followed by the removal of free ligand or alpha tocopherol in concentration of 0.5 microM. In the presence of PMA the inhibiting action of alpha-tocopherol was not found (under the 0.5 microM of alpha tocopherol) or was small (under the concentration of it 100 microM of alpha tocopherol). The addition of alpha-tocopherol in concentration of 0.5 microM and 50 microM caused the inhibition of blasttransformation of lymphocytes and activity of natural killer cells. This effect was expressed more under the low contents of this vitamin in the incubation medium. The level of blasstransformation of blood in vitamin E-deficient rats was by 20% less than in normal. The exogenous addition of alpha-tocopherol in concentration of 0.5 microM did not affect this value. It was suggested that vitamin E can affect the respiration burst of neutrophils, blasttransformation of lymphocytes and activity of natural killer cells and these actions depend on its concentrations. PMID- 9848161 TI - [Classification of results of studying blood plasma with laser correlation spectroscopy based on semiotics of preclinical and clinical states]. AB - The usage of laser correlation spectroscopy for verification of preclinical and clinical states is substantiated. Developed "semiotic" classifier for solving the problems of preclinical and clinical states is presented. The substantiation of biological algorithms as well as the mathematical support and software for the proposed classifier for the data of laser correlation spectroscopy of blood plasma are presented. PMID- 9848162 TI - [Change in urea-forming function of rat liver in the acute and recovery period after radiation exposure]. AB - The results of investigations of some metabolites and enzymes of ornithine cycle in total gamma-irradiated rats are presented. It is shown that ionizing radiation furthers the intensification of liver ammonium binding function. It took place without visible enzyme activation during the first period of defeat. The increasing of enzyme activity at the top of pathological process development is also shown. The definite dose-dependence was observed in this period. The month after irradiation the liver function was decreased and during two month after influence the liver function was not normal. PMID- 9848163 TI - [Effect of heavy metal ions on superprecipitation and ATPase activity of uterine smooth muscle actomyosin activity]. AB - The inhibiting effect of Pb2+, Zn2+ and Cd2+ on Mg(2+)-dependent superprecipitation and ATPase activity of myometrium actomyosin. The inhibiting effect of heavy metals cations on the both processes satisfies the succession: Pb2+ > Zn2+ > Cd2+. Cadmium and zinc ions in concentration of 1 mM stimulate the initial velocity (v0) of Mg(2+)-dependent superprecipitation by 25% and 80%, respectively, while the lead ions under the same concentration inhibit the initial velocity by 40%. It is possible that these results evidence for the direct effect of ions of heavy metals on active-myosin interaction (tested by v0). May be that the mechanisms of Cd2+ and Zn2+ action on the one hand and Pb2+, on the other hand, on the interaction of the contractile proteins of the uterus smooth muscle are different. Cations of Pb, Cd or Zn introduced to the incubation medium instead of Mg2+ (5 mM) also stimulate both superprecipitation and ATPase activity but the level of the both processes decreases by 65%, 20% and 5%, respectively, as compared to control (i.e. in presence of Mg2+). It is probable that the cadmium, zinc and lead cations can substitute magnesium ions in the active centre of myosine as well as in the sections of significance for the process of superprecipitation of actomyosine. At the same time the substitution is less efficient for realization of the superprecipitation reaction and ATP hydrolase process than when magnesium ions are available in the incubation medium. EDTA and EGTA do not remove the inhibiting effect of Pb2+, Cd2+ and Zn2+ on the contractile activity and ATP-hydrolase reaction of actomyosine complex of the uterus smooth muscle. The results obtained prove that the ions of heavy metals can effect the uterus smooth muscles at the stage of actin-myosine interaction. PMID- 9848164 TI - [Effect of Arnica montana on the state of lipid peroxidation and protective glutathione system of rat liver in experimental toxic hepatitis]. AB - Effects of Tinctura Arnica on lipids peroxidation and on the protective glutathions system of liver in rats in case of experimental toxic hepatitis have been studied. Toxic hepatitis is accompanied by deep alterations of the oxidant antioxidant status of the body. Intoxication of the body by CCl4 results in intensification of the free radicals formation particularly in liver: accumulation of lipids peroxidation molecular products, glutathione system enzyme activity inhibition in early terms and its partial restoration in remote terms has been seen. Our studies revealed that Arnica montana infusion inhibits the rate of lipids perioxidation products formation, affects the glutathione system enzymes activity. PMID- 9848165 TI - [Effect of aluminum poisoning and gamma-irradiation on the antioxidant system of the body and study of possibilities for correction]. AB - The influence of selenium and rutin on the antioxidant protection system of the organisms in connection with Al-intoxication and gamma-irradiation was investigated. It has been determined that selenium increases the antioxidant protection caused by aluminium. Under simultaneous action of aluminium and gamma irradiation upon the organism selenium stimulated accumulation of toxic lipids peroxide oxidation products. Rutin action in the situations under investigation caused weakening of antioxidant protection. PMID- 9848166 TI - [Detoxicating function of the placenta of childbearing women from ecologically unfavorable regions of the Ukraine]. AB - A study of placental detoxifying activity was undertaken using as the investigated indices glutathione transferase and glutathione reductase activities and total amount of SH-groups and those of low-molecular weight thiols. Malonic dialdehyde content was used as a measure of lipid peroxidation. The placental samples were obtained in several regions of the Ukraine judged either "clean", chemically polluted or radioactively contaminated. In the samples from the "clean" region glutathione transferase and glutathione reductase activities as well as the total amount of SH-groups were higher than corresponding indices in ecologically unfavorable regions of the Ukraine. At radioactively heavily exposed women low glutathoione transferase activity combined with low content of SH groups and pronounced increase of malonic dialdehyde. In the regions with the lower summary effective equivalent annual expositional doses the indices of placental glutathione status are more close to those characteristic for placenta from "clean" regions. The consequences of the more intensive but short exposure characteristic for the women evacuated from Pripyat on the second day after catastrophe were less manifest in the indices of the glutathione status. A decrease of glutathione transferase activity at chemically exposed women was in the range of the lowest values obtained for heavily radioactively exposed ones but differ by higher content of total SH-groups and low molecular weight thiols. The glutathione transferase activity of cytosol from the samples of chemically exposed women increased in vitro after their treatment with the reducing agent. More probable the different mechanisms underlie the similar reduce of glutahtione transferase activity in radioactively contaminated and chemically polluted regions of the Ukraine. The decreased activity of glutathione transferase activity in placenta correlated with the increased frequency of complications during the pregnancy and the delivery and with the worsening status of newborns. PMID- 9848167 TI - [Brain lipid composition of the rat with experimental morphine dependence]. AB - The brain phospholipid content was studied in rats with morphine physical dependence. It was found that level of total lipids, total phospholipids and cholesterol was diminished. The significant decrease of absolute amounts of phosphotidyl serine, phosphotidyl inositol, sphingomyelin and phosphatidyl choline was shown. At the same time the percent relation of individual phospholipids did not change. As the exclusion, the quantity of phosphatidyl ethanolamine plasmalogen lowered in rats with morphine dependence. The percent content of fatty acids 16:0, 18:1 omega 9, 20:1 omega 11, 20:4 omega 6 and 22:5 omega 3 was found to be reduced. At the same time the percent amount of i18:0 and 20:0 was shown to increase. As a result the unsaturation index dramatically diminished. It was also shown that under morphine physical dependence the acyl residues of phosphatidyl choline and phosphatidyl ethanolamine changed. It was concluded that lipids play significant role in the development of morphine physical dependence. PMID- 9848168 TI - [Activity of antioxidant enzymes and intensity of free radical oxidation processes in swine tissues during postnatal adaptation]. AB - It was studied the activity of superoxide dismutase, glutathione peroxidase, glutathione reductase, and contents of malondialdehyde and hydroperoxide lipids in the tissues of skeletal muscles, heart, lungs, testicles in the piglets of 1-, 3-, 5- and 10-days age. It was found, that tissues of 1-days piglets characterized by high contents of products of hydroperoxide lipids and activity of superoxide dismutase by low activity of glutathione peroxidase and glutathione reductase. The products contents of hydroperoxide lipids in piglets tissues in the period of 1 to 10-days age reduced. The activity of enzymes of antioxidant system depends from stage of postnatal adaptation and from the kinds of tissues. PMID- 9848169 TI - [Metabolic disturbances in blood from patients with senile cataracts accompanied by somatic diseases]. AB - The results of metabolic parameters determination, which characterize the state of protein's metabolism, membranes stability, cell s system of antioxidant deficiency at the patients with senile cataract are presented with the purpose to make clear the differences between biochemical mechanisms of lenses clouding progress at the elderly persons without and on the background of different somatic pathology. Our results have shown that disturbances of membranes structural-functional parameters and the lowering of activity glutathione metabolic system is more expressed at the patients with accompanying diseases of cardio-vascular system, gastro-intestinal tract, liver and chronic alcohol intoxication. The exhaustion of antioxidant system is observed at the patients with senile cataract in the cases of accompanying disease of liver and chronic alcohol intoxication unlike the patients without accompanying somatic diseases. Diseases of cardio-vascular's system, gastrointestinal tract and liver effort the metabolic disturbances in blood of the patients with senile cataract. PMID- 9848170 TI - [Automation of composite processes for biochemical studies]. AB - Approximate division of biochemical investigations into three components: experimental research, results treatment and modeling, as well as external information filling is made. Computerization of biochemical researches for these components are analyzed and authors developments for experimental researches computerization on IBM-compatible computer basis are presented, as well as the local computer net of biochemistry institute. PMID- 9848171 TI - [Metabolism of methionine in tissues of cattle in vitro]. AB - After incubation of homogenates of liver, kidney, brain, skeletal muscles mucosa of rumen and empty intestine of cattle in Krebs-Ringer's phosphate buffer with [2 14C]methionine 25.8-43.4% radioactive label was found in protein; 28.1-42.0%, in CO2; 12.0-18.6%, in glucose; 8.2-14.9%, in glycogen; 3.1-6.3%, in lipids. The highest level of [2-14C]methionine utilization in protein synthesis was found in skeletal muscle; in energetical processes, in liver; in the process of gluconeogenesis, in skeletal muscles and liver; in lipid synthesis, in brain; the lowest level of [2-14C]methionine utilisation in above-mentioned process was found in skin. PMID- 9848172 TI - [Latent ceruloplasmin activity in the frontal lobe of albino rat brains]. AB - Latent ceruloplasmin activity was revealed in 5% of homogenates of the frontal lobes of the albino rats' brain. While assessing the activity of this enzyme in the investigated assays in the dynamics of their storage at 4-8 degrees C, the value under study continuously increased and on the 14th day of the experiment it reached 137% of the initial values. 1-Triphenylphosphoniomethylnafthalene bromide did not exert any effect on the dynamics of changes of ceruloplasmin activity in biological assays. PMID- 9848174 TI - [Effect of nutritional additives on oxidative processes in rat liver mitochondria]. AB - The paper deals with the influence of sweeteners and aromatic compounds to the oxidative phosphorylation in rat liver mitochondria. The effect of food additives depends from its concentrations and time action. PMID- 9848173 TI - [Free radical oxidation of lipids and antioxidant system of blood in infertile women in a radioactive environment]. AB - Under the influence of different external factors, much attention was given to the study of the characteristics of pro- and antioxidant systems in the organism. It was studied the state of lipid peroxide oxidation in infertile women, who were evacuated from Chernobyl and Pripyati, and those who permanently live in the radiational-fected area. An appreciably high activity of lipid peroxidation was found in women who were affected by the radioactivity of different intensity. A sharp decrease in the strength of antioxidant defence system promotes a marked activation of the lipid peroxide oxidation. The cause of the sharp decrease of the intensity of antioxidant defence system is its exhaustion. PMID- 9848175 TI - [Physiological and adaptive value of Na+,K+-ATPase isoenzymes]. AB - A current state of the researches related to functional specificity of the Na+, K(+)-ATP-ase isozymes determined by their catalytic properties, individual sensitivity to regulation by hormones and other endogenic substances and to membrane structural disorders has been reviewed to clarify the physiological expediency of the Na+, K(+)-ATP-ase isoforms. The data on isozymes expression in brain in the ontogenesis, their individual sensitivity to ouabain and peculiarities of the membrane properties are considered. The adaptive significance of the Na+, K(+)-ATP-ase isozymes in expansion of tissue compensatory resources at some physiological and pathological states of organism is supposed. PMID- 9848176 TI - [Structure and metabolism of collagens in various pathologies]. AB - Collagen constitutes approximately one third of the body's proteins; it has different functions, and changes in its metabolism and structure accompany almost every pathologic process. This article reviews basic knowledge of the alterations and defects in collagen metabolism and structure which give rise to a number of heritable and acquired diseases, their causes and consequences. PMID- 9848177 TI - [Protein motors: structure and generation of mechanical force]. AB - Recent data about the structural and functional features of protein motors (myosin, dynein, kinesin) are presented. The mechanism by which the chemical energy of ATP is transduced into mechanical work is discussed. PMID- 9848178 TI - [Effect of preincubation on smooth muscle myosin light chain kinase activity]. AB - Phosphorylation of myosin regulatory light chain (RLC) catalysed by myosin light chain kinase (MLCK) is a key reaction in the regulation of actin-myosin interaction in smooth muscle. The activation of MLCK by calmodulin (CaM) and Ca2+ was investigated over a wide range of the enzyme concentrations using myosin or its RLC with Mw = 20 kDa as substrates. Kinase activation by CaM (at saturating Ca2+ concentrations) was characterized by positive cooperativity even though noncooperative activation would be expected from the established 1:1 binding stoichiometry between MLCK and CaM. The activation of the kinase by Ca2+ was also cooperative but only at relatively low CaM levels. This cooperativity was shown to result from time dependent changes in MLCK that take place during its incubation with Ca2+ and CaM before substrate addition in phosphorylation assays. As a result the kinase activity as a function of its concentration at constant CaM level was biphasic: there was the activity optimum at 1:1 ratio of CaM to MLCK and almost complete inhibition at 3 to 7 molar excess of kinase over CaM. Such changes that take place during 10 to 15 min preincubation with Ca2+ and CaM may involve the kinase supramolecular structure formation or/and its conformational rearrangements. PMID- 9848179 TI - [Conformational modifications of smooth muscle light chain kinase]. AB - Our recent investigations have shown that smooth muscle myosin light chain kinase (MLCK) exists in solution as a mixture of oligomeric, dimeric and monomeric species; besides during preincubation (maintaining of the activated enzyme without substrate) with substoichiometric amounts of calmodulin (CaM) it undergoes definite changes leading to several fold lowering of its activity. Fluorescent data obtained in this work suggest that such kinase inhibition must not be connected with quantitative redistribution of different kinase species but rather it is the result of conformational modifications of this enzyme activated molecules leading to the reduction of their affinity to CaM. Such conformational rearrangements took place also at equimolar kinase to CaM ratio (or CaM excess) but in this case they were characterized by lower depth and insignificant MLCK activity fall. The nature of these conformational changes is discussed. PMID- 9848180 TI - [A new method of preparing and some properties of high molecular weight monoamine oxidase from swine liver mitochondria]. AB - Isolation of highly purified and highly molecular monoamine oxidase (MAO) from pig liver mitochondria have been worked out. Specific activity of isolated preparation is 2700 times higher than of original mitochondria homogenate. Enzyme solubilization by digitonin, affinity chromatography purification and ultrafiltration underlie this method. MAO catalytic properties changing during the process of purification by different methods have been investigated. Substrate specificity was studied; kinetic parameters of enzymatic desemination were calculated. PMID- 9848181 TI - [Study of sorption properties of organosilanes to be used as the basis for hemosorbents and diagnostic enzyme and immunosensors]. AB - The abilities of different types of organosilanes, in particular of polymers with: 1) completely or 2) partially hydrophobical surfaces; 3) regular changes of part of silicium ions by metal ions; 4) preliminary aminosilanization were studied for sorption of ammonia ions, urea, cholesterol, creatinine, albumin, IgG, haemoglobin and myoglobin. Polymethylsiloxane was used as haemosorbent for directed sorption of myoglobin and haemoglobin from solution and blood. It didn't hemolysate red cells. The high efficiency of those organosilanes for sorption of haemoproteins it was shown. Organosilanes were very good as membrane for immobilization of urease and IgG-specific antibodies to create enzyme sensor and immunosensor based on the ionsensitive field effect transistors. The advantages and possibilities of organosilane usage as haemosorbents in the field of medicine of catastrophes as well as for sensor technology are discussed. PMID- 9848183 TI - [Kinetics of conformational changes of methemoglobin complexed with liposomes]. AB - Kinetics of methemoglobin structural changes in the complex with liposomes composed of phosphatidylcholine and its mixtures with cardiolipin has been studied. The amplitudes and rate constants of the two observed kinetic phases are determined. The fast kinetic phase is attributed to the formation of the unstable intermediate protein form, while the slow one is assumed to reflect dissociation of the heme--globin complex. PMID- 9848182 TI - [Chromatography of thrombin and antithrombin III on biospecific silochromes]. AB - It was shown that the seaweed sulfopolysaccharides Furcellaria and Phyllophora- furcellaran and phyllophoran--are the competitive inhibitors of thrombin-induced fibrinogen clotting. Biospecific adsorbents for thrombin and antithrombin III (AT III) isolation were synthesized with heparin, phyllophoran and furcellaran as ligands on aminopropylsilochrome and this proteins sorption characteristics on this sorbents in comparisons with heparin-cellulose and heparin-sepharose were investigated. Possible application of the synthesized adsorbents both for thrombin, AT III purification from blood products and for alpha-thrombin, beta and gamma-thrombin separation are discussed. PMID- 9848184 TI - Adenosine improves cardiomyocyte respiratory efficiency. AB - The role of adenosine on the regulation of mitochondrial function has been studied. In order to evaluate this the following experiments were done in isolated rat cardiomyocites and mitochondria using polarographic techniques. Cardiomyocyte oxygen consumption (MVO2) and mitochondrial respiratory function (State 3 and State 4, respiratory control index, and ADP/O ratio) were evaluated after exposure to adenosine. Cardiomyocyte MVO2 was significantly lower in cells previously exposed to adenosine (10 microM, 15 min or 30 min cell incubation) than in cells not exposed to adenosine (control). Addition of dipyridamole (10 microM) or 8-(p-Sulfophenyl) theophylline (50 microM) to cardiomyocytes before adenosine incubation prevented the adenosine-induced changes in MVO2. Mitochondria obtained from isolated perfused beating heart previously perfused with adenosine (10 microM, 30 min heart perfusion) also resulted in significant increases in ADP/O and respiratory control index compared to matching control. Mitochondria isolated from cardiomyocytes previously exposed to adenosine (10 microM, 15 min or 30 min cell incubation) resulted in a significant increase in mitochondrial ADP/O ratio compared to control. Adenosine-induced decrease in cardiomyocyte MVO2 may be related to an increase in efficiency of mitochondrial oxidative phosphorylation, and more economical use of oxygen, which is necessary for survival under ischemic stress. PMID- 9848185 TI - [Comparative analysis of semiotic shifts, established by LCS of blood plasma from random samples of studied subjects from the zone of the Chernobyl accident, "Ural Radiation Trace", and collaborators from St. Petersburg Institute of Nuclear Physics of the Russian Academy of Sciences]. AB - Using the developed "semiotic" classifier of laser correlation spectra of blood plasma the authors have carried out the verification of organism states of patients from the zone of Chernobyl accident, "Ural radiation trace" and collaborators from Sanct-Petersbourg Institute of Nuclear Physics. An analysis of results obtained using accidental selections which differed as to the character of radiation injury evidences for high informativeness of "semiotic" classifier of laser correlation spectra of blood plasma. PMID- 9848186 TI - [Effect of "Naftusia" water on some indicators of metabolism in rats]. AB - It is shown that giving rats natural water "Naftusya" to drink during 5 days initiated a complex of variations in change of substances, conditioned by general adaptational reaction. Ultraviolet radiation of water has no effect on its balneoactivity while ultrafiltration attenuates it a little. PMID- 9848187 TI - [Effect of hyperbaric oxygenation of lipid peroxidation and the antioxidant system in liver of white rats with acute sodium nitrate poisoning]. AB - The experimental study carried out with white rats Wistar trend, which were introduced of sodium nitrate at a rate of 9.6 g/kg of their mass. It follows to the development of considerable activation of lipid peroxidation and depression of antioxidant system in liver. The obtained results permit supposing the significant role of nitric oxide (NO) in liver as a factor resulting in accumulation of peroxidation products. The research has stated that the use hyperbaric oxygenation (HBO) prevents considerable activation of lipid peroxidation and the decrease of antioxidant enzyme (catalase and superoxide dismutase) activity. The results permit supposing that the effect HBO is connected with the decrease of the rate of reduction of nitrate-ions to NO. PMID- 9848188 TI - [Biochemical mechanisms of the toxic effect of oxalates]. AB - The effect of oxalic acid and its salts on the biochemical systems of the homeostasis of white rats were investigated. It has been shown that oxalates inhibit enzymes of energy metabolism and cause the changes in parameters of nephrotoxicity, electrolyte balance of blood plasma and urine, lipid peroxidation, antiperoxidational defence system. PMID- 9848189 TI - [Urea-forming function of the liver in aging rats]. AB - The investigations of some metabolites and enzymes of ornithine cycle were performed for infant and adult rats. The decreasing of enzyme activity and the decreasing of urea production have been shown as a reflection of anabolic processes prevalence for a growing organism. PMID- 9848190 TI - [Regulatory components of the eukaryotic translation apparatus in normal states and after exposure to ionizing radiation]. AB - The comparative analysis of content prostaglandins E2, F2 alpha and cAMP, cGMP in the compositions of the high-molecular aminoacyl-tRNA synthetase complexes and endoplasmic reticulum vesicules from the rat liver has been carried out. The influence of lethal and sublethal X-ray doses on the levels of cyclic nucleotides and prostaglandins has been studied. Cyclic nucleotides and prostaglandins proved to be rapid and high sensitive transmitters of ionizing radiation influence on the eukaryotic translation apparatus. PMID- 9848191 TI - [Effect of electron irradiation on the structure of microsomal membrane proteins]. AB - Using tryptophan fluorescence quenching by iodide and acrylamide the effect of electrons with the energy 5 Mev on the structure of microsomal membrane proteins has been studied. Conformation of microsomal proteins were found to change upon irradiation. PMID- 9848192 TI - [Structural-functional changes in membranes and external perimembrane layers of erythrocytes in hyperepidermopoiesis]. AB - The coat properties of erythrocytes in hyperepidermopoesis had been studied using the vital quantitative method of the cell coat (outer perimembraneous layer) identification with alcian blue (AB). It is shown that the stage sharpening in psoriasis was accompanied an increase in AB sorption by erythrocytes for 13.2. The qualitative and quantitative gangliosidic spectrum of erythrocytes and serum does change according to the date of chromatography in hyperepidermopoesis. The role of this changes are discussed in pathogenesis of the given disease. PMID- 9848193 TI - [Dynamics of the interaction of monoclonal antibodies with antigens immobilized on plates]. AB - A new method to determine a rate constant and total reaction product using data of reaction product kinetics is proposed. The method is based on the solution of the system of irrational equations (for special case), which describes such processes. This method was used to determine the reaction rate constant of interaction of some monoclonal antibodies with the antigens immobilized on immunological plates. PMID- 9848194 TI - [Antiradical and antioxidant activity of certain membranotropic substances of synthetic and plant origin]. AB - Antiradical and antioxidative activities of dieton, phoridon, niphedipin, phlamicar, mildronat in vitro were studied by spectrophotometer's method with the use of dyfenilpycrilgydrasil and method of pulse voltametry. Also influence of drugs was studied under antioxidative insufficiency (AOI), which had modelled by exclusion of alpha-tocopherol from the animal's ration. PMID- 9848195 TI - [Role of inositol triphosphate in the formation of the calcium signal in T lymphocytes]. AB - The mechanisms of TCR/CD3 complex and phospholipase C interaction, inositol-1,4,5 triphosphate role in both initial transient and sustained elevation of intracellular Ca2+ and in calcium oscillations driving following T cells stimulation are discussed. Data on IP3-receptors structure and localisation in lymphocytes are summarized. PMID- 9848196 TI - [Basic (cleaving arginine and lysine residues) metallocarboxypeptidase of mammalian tissue: structure, properties and function]. AB - The structure, physical, chemical and catalitical properties, functions and biological role of mammalian basic carboxypeptidases are observing. On the strength of the genetic and filogenic research data it is supposed the existence of a family of basic metal-dependent carboxypeptidases and the plan of it evolution is proposed. The connections between structure, localisation and function of this enzyme are discussed. PMID- 9848197 TI - [Primary hypercholesterolemia: mechanisms of its development in man]. AB - The review is devoted to hypercholesterolemia which is one of the leading risk factors for IHD. By its origin it can be primary and secondary. Three major mechanisms of primary hypercholesterolemia in humans are discussed. They are: low activity of LDL receptors, reduced affinity of LAL for receptors and overproduction of lipoproteins containing apo B. Depending on mechanism nonidentical molecular defects leading to cholesterol imbalance in cells or in circulating lipoproteins are occurred. The understanding of the nature and mechanisms of hypercholesterolemia development is of great clinical value, because having determined molecular defect and using drugs combinations the majority of patients are a success to have their lipids normal. Only in the case of homozygous familial hypercholesterolemia medicamentous treatment becomes secondary and the principal therapeutic methods are plasmapheresis or selective LDL apheresis. Gene therapy as a method of homozygous familial hypercholesterolemia correction is in forthcoming future. PMID- 9848198 TI - [Ion-sensitive endogenous regulators of Na+,K+-ATPase obtained from epithelium of rat small intestine]. AB - From epithelial layer of rat intestinal were selected water soluble substances which influenced on Na+,K(+)-ATPase activity in 2 different ways: substances with molecular weight 220 Da, 400 Da were its activators, substance with weight 150 Da inhibitor. Na+,K(+)-ATPase activators preincubated previously with Na+ acquire properties of inhibitors. Bivalent cations Ca, Mg remove this effect of Na-ions. PMID- 9848199 TI - [Temperature dependence of the transmembrane distribution of non-electrolytes in human erythrocytes]. AB - Influence of temperature on the behaviour of non-electrolytes n-alcohol series distribution between intra- and extracellular environment of human red blood cells was studied by means of 1H-NMR technique. It was shown, that the temperature dependences of the distribution coefficients Q of the non electrolytes solutions between erythrocytes and medium in the range of T = +4( )+37 degrees C could not be describe by Arrhenius equation. There was supposed that the absorbtion-phase theory may be used for the description of non electrolyte absorbtion and non-electrolyte distribution between human erythrocytes and intercellular medium. PMID- 9848200 TI - [Features of the organization of the cytochrome systems and oxygen regimen in skeletal muscles of marine fishes]. AB - The organization of cytochrome systems in pelagic (Mugil so-iuy) and benthic (Platichthys flesus luscus) species of sea fishes were investigated. The total cytochrome content was 3.6-3.7 times higher (p < 0.001) in muscles of active M. so-iuy and made 67.5 +/- 5.3 nM g-1 for red and 5.91 +/- 0.58 nM g-1 for white muscle tissue. Cytochrome stechiometry had non-compensation character in benthic P. flesus luscus. Cytochrome ratio-b/aa3, was less than one and was equivalent 0.80-0.83 for both muscle groups. On the contrary it was higher than one--1.47 1.66 (p < 0.001) in M. so-iuy. Differences in cytochrome organization were connected with oxygen state of muscle tissue of these fishes. Mean oxygen tension in P. flesus luscus muscles was lower for 20.1-22.5% (p < 0.001). A number of anoxic and hypoxic zones (less than 6 mm Hg) in muscle tissue of this fish made 70 and 4-6% for white and red muscles accordingly. This value was 1.7-2.5 times lower in M. so-iuy muscles. PMID- 9848201 TI - [Antiradical activity and membranotropic action of cardio- and vasotropic compounds of varying chemical nature]. AB - Antiradical activity and ability to interaction with phosphatidylcholine bilayer of the physiological active compounds (PhAC), concerning the classes of phenylalkilamines (dobutamine, verapamil), dihydro pyridines (BAY-K-4688, nifedipine), analogues of crown ether (carbicyle, diol) was studied. By means of the method of microcalorimetry and spectrophotometry it was shown the complexing ability PhAC with the phospholipid bilayer of the model membrane. It was stated the simbasity in the changes of the thermal effect of the compounds with the negative inotropic activity during its reaction with phosphatidylcholine bilayer and antiradical activity. That show on the presence of antiradical component in the mechanism that compounds' action. PMID- 9848202 TI - [State of antioxidant and oxygen transport system of the blood in the process of adaptation of the body to hypoxic hypoxia]. AB - This article shows that in oxygen transported and antioxidative systems considerable changes take place during dynamic of adaptation to oxygen deficiency of inhaled air (every day 4 hours stay of animals in barochamber under pO2 7.45 kPa during 12 days). Single influence of hypoxic factor induced in erythrocytes peripheral blood increasing of haemoglobin affinity to oxygen, decreasing SOD and catalase activity on the background of increasing of malon aldehyde level. On 12 th day of adaptation of animals contents of TBA-active products in erythrocytes is saved on high level, activity of antioxidative enzymes is approached to original meanings, haemoglobin affinity to oxygen is decreased in comparison with control. Detected increasing content of creatine in erythrocytes during dynamic of rat adaptation to hypoxic hypoxia witnessed about erythropoiesis activation and quickened exit out of marrow young forms of erythrocytes. PMID- 9848203 TI - [Effects of modifications of lipoproteins by water soluble forms of lineoleic hydroxamic acid on biochemical markers of development of atherosclerosis]. AB - Effects of water-soluble form of linoleic-hydroxamic acid, inhibitor of lipoxygenase, on process of atherogenesis (alimentary model) were studied at rabbits. It was shown that inhibition of activity of lipoxygenase during process of development of atherosclerosis led to considerable decrease of area of lipoid plaques and lipoidosis, decreased content of cholesterol in blood and improved free-radical modification of lipoproteins of blood. PMID- 9848204 TI - [Changes in lipid metabolism during oxidative stress caused by cobalt chloride injection]. AB - The cobalt chloride influence on rat liver and serum blood lipids and lipoproteins content and composition was investigated. The data obtained show that the cobalt chloride injection leads to the oxidative stress development and to activation of lipid transport and metabolism. It has been shown that there is system of lipids homeostasis in the organism and it is activated under conditions of oxidative stress. The system blood lipoproteins and liver lipoproteins was found to participate in metabolism adaptation on oxidative stress and in maintenance of biological membranes structure and functioning. PMID- 9848205 TI - [Antioxidant status in manganese poisoning]. AB - Pigs intoxication by chloride manganese in dose 50 mg/kg perk day for 10 days brings to lowering antioxidant protection in blood and the level of respiratory neutrophils' burst. In liver and kidneys superoxide dismutase activity increases, in liver--increasing of initiate and final malondialdehyde contents and lowering of concentration of restored glutathione and ATP. PMID- 9848206 TI - [Content of components of the renin-angiotensin-aldosterone system of blood plasma of healthy children in the early neonatal period]. AB - The dynamics of the renin and angiotensin converting ferment activity and of the aldosterone concentration in the plasma of blood by healthy newborn full-term in the first week of the life after the birth was determined. The normative values of the studied indexes were obtained. The researches have shown the considerable functional tension of the renin-angiotensin-aldosterone system, that has a very important signification for optimal course of adaptation and anabolic processes by the children at the very beginning of the postnatal development. PMID- 9848207 TI - [Activity of elastase and its inhibitors in arterial tissues and blood serum under experimental arterio-atherosclerosis]. AB - The role of the elastolytic system serum and arterial tissues in pathogenesis of vascular pathology was investigated on rabbits in experimental arterio atherosclerosis. Activation of proteolytic enzymes, in particular elastase, sharp alteration capacity of inhibitors in arterial wall both at Menckeberg arteriosclerosis and atherosclerosis, permits to assume, that in the basis these different pathologic processes can lie general mechanisms, which will be realized through disturbance of the elastolytic system balance in blood vessels. PMID- 9848208 TI - [Formation of inositol triphosphate in lymphocytes of rat spleen in the early stages after x-irradiation]. AB - Stimulation of 32P-phosphoinositides turnover and phosphatidylinositol-4,5 biphosphate hydrolysis is observed 6 h after animals X-ray irradiation in a dose 1 Gy. Both mitogenic lectins LPS and PHA (10 mg/ml), but not 0.5 microM A23187 induce inositol phosphates release from endogenous [3H]inositol--phospholipids in lymphocytes. The level of inositol triphosphate is increased within 1-6 h and decreased in 12 h after irradiation. 45Ca2+ influx through lymphocytes plasma membrane is not changed within 3 h, but increases greatly 12 h after exposure. Data obtained indicate that IP3 formation is an early signal for radiation induced changes in calcium homeostasis. PMID- 9848209 TI - [Effect of imipramine and ftoracizin on platelet aggregation and smooth muscle contraction of the ureter]. AB - The influence of the tricyclic antidepressants imipramine and ftoracizin on platelet aggregation and smooth muscle contractility was investigated in comparison with action of known smooth muscle relaxant and platelet aggregation inhibitor, papaverine. It has been shown that the tricyclic antidepressants possess potent spasmolytic activity but unlike papaverine have no effect on platelet aggregation. The biochemical mechanisms of the non-specific action of tricyclic antidepressants as well as some other structurally related-drugs are discussed. PMID- 9848210 TI - [Effect of irradiation on the degradation of rat thymocyte chromatin]. AB - Genome instability of adaptive nature is formed under the experimental influence on a cell. Under critical conditions, strategy of organism is to damage the cells that cannot be restored and controlled by including the program of apoptosis. The ordered internucleosomal DNA degradation is considered to be one of the proof attributes of immunocompetent cell apoptosis. We investigated the effects of various doses of irradiation on the thymocytes chromatine fragmentation in 1,2,3 hours after a single X-ray exposure or after chronic influence in conditions of Chernobyl research base. By the means of electrophoresis in agarose and judging by polydeoxyribonucleotides accumulation we observed the "ladder pattern" of degradation in 3 hr after single 1 Gr irradiation (the smallest dose displaying the effect). We suppose that the influence of both chronic low-intensity irradiation taking place in Chernobyl and single X-ray exposure result in intensifying of DNA fragmentation in the cells of immunocompetent organs. PMID- 9848211 TI - [Effect of tocopherol deficiency and additional administration of tocopherol, dibunol and disodium selenite on enzyme activity in rat liver, which take part in the biotransformation of nitrosodimethylamine]. AB - The alimentary tocopherol deficiency is accompanied by decreased hydroxylase, demethylase, NADH- and NADPH-reductase, aldehyde dehydrogenase, arylesterase and glutathione reductase activity in rat's liver. It decreased the reduced glutathione and increased it's oxidized form concentration in the tocopherol deficient animals. The stability of microsomal membrane is decreased to solubilizing action of deoxycholate and trypsin. This changes, possibly, caused elevation of alteration of function enzyme's and microsomal membrane after nitrosodimethylamine (NDMA) administration in deficient rats. The 7-days injection of tocopherol (20 and 100 mg/kg), dibunol (80 mg/kg), sodium selenite (30 mkg/kg) increased aldehyde dehydrogenase, esterase, glutathione-dependent enzymes activity and increased of reduced glutathione concentration in liver, suppressed lipid peroxidation and increased survival rats after lethal dose carcinogen treatment. Supplementation of tocopherol decreased harmful action of nitrosodimethylamine on microsomal membrane and enzymes activity. PMID- 9848213 TI - [Inhibiting effect of the aqueous extract of Arctostaphylos uva-ursi on myrosinase activity from Brassica napus seeds]. AB - The search of inhibitors of the myrosinase enzyme and also enzymes able to transform the glucosinolates into non-toxic combinations has been carried out among the water extracts of the plants row. The inhibitor activity of the water extract of Arctostaphylos uva-ursi and its foresee acting outset-hydroquinone have been discovered. The direct dependence of the glucosinolates decompose degree from concentration of hydroquinone and quinone has been determined. The cultivation of Brassica napus cut seeds by quinone in correlation 1:100 (m:m) stopped the glucosinolates decomposition by endogenous myrosinase for 50%. PMID- 9848212 TI - [The sensitivity of catalytic subunit isoforms of Na+,K+-ATPase of rat brain to peroxide membrane modification]. AB - The comparative study of the sensitivity of Na+, K(+)-ATPase isozymes from cerebral cortex to ascorbate-dependent membrane peroxidation was conducted. With highly inactivated Na+, K(+)-ATPase the degree of inactivation of the SH dependent ouabain-sensitive forms alpha+ (alpha 2 and alpha 3) is higher than glycoside-resistant isoform alpha 1. The process is accompanied by simultaneous lipid peroxidation and decrease of SH-groups amount in enzyme preparations. The combined nature of the oxidative Na+, K(+)-ATPase inactivation, accompanied by the direct oxidation of enzyme SH-groups and modification of lipid environment is supposed. PMID- 9848215 TI - [Effect of contrical on proteolytic enzymes of blood serum in acute emotional pain stress]. AB - General proteolytic and antitryptic blood serum activity in acute emotionally painful stress was investigated as well as that on the background of the before injected contrical. It was determined that in case of a stress there increases the general proteolytic activity of blood serum and at the same time antiproteolytic activity decreases. The coefficient of the relation of antiproteolytic activity to proteolytic activity of blood serum in case of a stress may become of 50% of the control. The introduction of inhibitor of proteinase contrical before the beginning of stressor influence promotes the decrease of general proteolytic activity to the level of the control indexes. PMID- 9848214 TI - [Relationship between the oxidation-reduction system of astrocytes with production of active forms of oxygen]. AB - Cells of neuroglia--the astrocytes are of interest from the point of view of their participation in phagocytosis. Phagocyte ability to generate active oxygen forms (AOF) as used as the basic criterion of the estimation of their functional activity. For the purpose to clear up molecular and cellular mechanisms of phagocytosis a study of astrocyte redox-systems, participating in production of AOF, was undertaken. Registration of AOF in astrocytes was carried out using a method of luminol-dependent chemiluminescence. Primary culture of guinea pig astrocytes was used. Spontaneous chemiluminescence of low intensity was found for the astrocytes at the presence of luminol. The destruction of the cells was accompanied by a significant growth of the intensity of spontaneous chemiluminescence. Suspension of endocutosis inductors, particle of latex and phytohemagglutinin, added to astrocytes did not result in formation of AOF, characteristic for other cells, possessing phagocytosis. It was established, that addition of hydrogen peroxide destroys astrocytes at the presence of luminol and gives rise to the emission. Chemiluminescence was not observed in similar experiments with intact cells. A conclusion was made that inside astrocytes there are structures, which show peroxidase-like activity. PMID- 9848216 TI - [Status of the system of hemostasis in patients with non-insulin-dependent and insulin-dependent diabetes mellitus without clinical manifestation of vessel pathology]. AB - Analysis of 17 haemostasis indexes has been done in 10 insulin-dependent diabetes mellitus patients, 5 insulin-independent diabetes mellitus patients and in 9 healthy donors. It was determined some of the same type changes and peculiarities which must be taken into consideration both by the estimation of blood system coagulation activity of patients treatment and by definition of disease prognosis etc. PMID- 9848217 TI - [Disorders of the processes of protein biosynthesis and a decrease in the content of cytochromes P-450 and b5 in the microsomal fraction of rat kidney in the dynamics of development of acute pyelonephritis]. AB - Noticeable decrease in microsomal protein level MOS terminal component- cytochrome P-450 and MOS median link--cytochrome b5 contents in microsomal fraction implicated in renal pathologic process was established in development of acute unilateral pyelonephritis. These findings reduction expression directly depended on severity and pathologic process development degree. One of the main causes led to reduction in cytochromes P-450 and b5 level and inhibition proteins biosynthesis may be influence of toxic products which are generated in substance exchange involved in prevalence of catabolic processes in injured kidney and intensification in endogenous intoxication syndrome on renal structures. Implication of many other factors effecting decrease in microsomal hemoproteins and protein levels in the damaged kidney in above pathology showed necessity to perform further investigations in the field given. PMID- 9848218 TI - [Delayed evaluation of urea content in bioassays using a chemical stabilizer]. AB - We have studied 126 chemical compounds. Moreover 9-triphenylphosphoniomethyl phenanthrene chloride has been found to stabilize bioassays and preserve their urea content unchanged for a period of 90-990 days. Due to its use a delayed technique of a quantitative assessment of urea in biological objects has been elaborated characterized by universal novelty. It may be used with the aim of studying metabolic processes in the cosmonauts' body, atomic submarine crews, members of polar, alpine, desert, underwater, space and other expeditions which cannot be accompanied by a biochemical laboratory. PMID- 9848219 TI - Pre-transplant donor-specific transfusions induce allograft rejection and IL-2 gene expression in the WKY-->F344 functional tolerance model of rat lung transplantation. AB - Our previous studies have shown that a spontaneous functional tolerance develops in a rat model of lung transplantation (WKY-->F344). The tolerance observed in this model may be due to the minor histocompatible differences in this combination, however, the possibility of a tolerogenic effect related specifically to the lung allograft must be considered. To further examine this model, the effect of pre-transplant donor-specific spleen cell transfusions (DSTs) was examined on the functional tolerance state seen in this model. F344 rats received WKY spleen cells on days -45 and -30 before lung transplantations. Control F344 rats received lung transplants without DSTs. Recipients in both groups were killed on day 7, 14, 21 and 49 post-transplant, and allograft rejection (AR) was graded histologically (stage 0-IV). Intragraft cytokine gene transcripts were examined on day 7 and 14 post-transplant using reverse transcriptase-polymerase chain reaction (RT-PCR) techniques to investigate the underlying immunological events occurring in each group. In addition, allogeneic (WKY) and third party (BN) skin grafts were placed on lung recipients at day 35 post-transplant to evaluate the development of systemic tolerance. It was seen that control animals showed moderate to severe lymphocytic infiltrations (stage II-III AR) in the first 3 weeks followed by spontaneous recovery with stage I-II AR on day 49. In marked contrast, DST-treated animals showed more aggressive AR with severe lymphocytic infiltration and haemorrhagic infarction (stage III-IV AR) by day 14-21, without any evidence of recovery on day 49. WKY skin grafts showed prolonged survival in control animals, but were promptly rejected in DST treated animals. Intragraft cytokine gene expression in control animals was characterized by no or weak expression of IL-2 and high IL-10, while DST-treated animals showed high levels of IL-2 transcripts. IL-2:IL-10 and IL-2:IL-4 ratios were significantly increased in DST-treated animals compared with controls on day 7 post-transplant. It was concluded that pre-transplant DSTs did not enhance allograft survival, but actually induced AR and ablated any immunological benefit of the lung allograft on induction of tolerance in the WKY-->F344 lung transplant model. It was found that the DST-induced AR was associated with a deviation of cytokine immune responses from a predominant Th2 to Th1 profile characterized by increased IL-2 gene expression in the allografts. We also conclude that factors other than the degree of histocompatibility matching, such as the route and timing of alloantigen exposure, and the amount or nature of alloantigens associated specifically with lung allografts, are involved in deviating native immune responses toward acceptance or rejection of lung allografts in this model of lung transplantation. PMID- 9848220 TI - An alternative conditioning regimen for induction of specific skin graft tolerance across full major histocompatibility complex barriers. AB - Previously, we developed a well-tolerated single-day protocol for induction of stable multilineage chimerism and permanent donor-specific tolerance across major histocompatibility complex (MHC) barriers, with preservation of the host's normal immune responses. In our murine model, recipient mice were treated with a single dose of anti-CD3, anti-CD4, low dose total body irradiation (TBI; 3-6 Gy) and allogeneic bone marrow cells. An alternative cytoreductive strategy that is well recognized in bone marrow transplantation, but has not been evaluated extensively in organ allograft recipients, involves the use of a combined chemotherapeutic drug treatment. The present data show that conditioning with low dose TBI, in a MHC-disparate donor-recipient combination, can be successfully substituted by a combined single low-dose dimethyl myleran (DMM)/cyclophosphamide (CY) therapy, resulting in both stable, mixed chimerism and specific skin graft tolerance. PMID- 9848221 TI - Kinetics of circulating cytotoxic T lymphocyte precursors that have a high avidity for donor antigens: correlation with the rejection status of the human cardiac allograft. AB - Studies on graft infiltrating cells demonstrated that accumulation of cytotoxic T lymphocytes (CTL) with high avidity for donor antigens (Ag) coincided with acute cardiac rejection. In the present study, we analyse whether such high-avidity CTL are present within the peripheral blood of cardiac transplant recipients and whether their kinetics correspond with the rejection status of the allograft. Using limiting dilution analysis (LDA), donor-specific CTL were enumerated in serial blood samples of seven patients. From each patient, 7-11 samples were obtained during the first year after transplantation and up to three samples were obtained at a later date. Enumerated donor-specific CTL were divided into CTL with high or low avidity for donor Ag, depending on their sensitivity to CD8 blocking. In contrast to the situation in the graft, the donor-specific CTL present within the peripheral blood were CTL precursors (pCTL) and not fully mature CTL (cCTL). The number of donor-specific pCTL among peripheral blood cells fluctuated irrespective of the rejection grade of the allograft, indicating that the frequency of circulating donor-specific CTL does not reflect the immunological status of the allograft. During acute cardiac rejection, 66% (median) of the circulating donor-specific pCTL had a high avidity for donor Ag. This percentage significantly exceeded pre- and postrejection values obtained during the first year post-transplantation (median, 39% and 37%, respectively). The disparity in avidity increased even further more than 1 year after transplantation, when stable engraftment was achieved. Among donor-specific pCTL in peripheral blood, those with a high avidity were absent (median, 0%). Hence the avidity of circulating donor-specific CTL might inform us about the immune status of the cardiac allograft. PMID- 9848222 TI - Fate of orthotopic corneal allografts in C57BL/6 mice. AB - Our aim was to determine whether corneal allografts placed in normal eyes of C57BL/6 mice enjoy immune privilege, and whether recipients of long-standing grafts acquire donor-specific ACAID. C57BL/6 mice received corneal grafts from BALB/c, C57BL/6, C3H, B10.D2 or BALB.B mice; for comparison, BALB/c mice received C3H or C57BL/6 corneal allografts. Delayed hypersensitivity and anterior chamber associated immune deviation (ACAID) induction were assessed in recipient mice at four and eight weeks after grafting. It was found that C57BL/6 mice resemble BALB/c mice in mounting more vigorous rejection reactions against minor H, rather than MHC, alloantigenic corneas, and in the acquisition of donor-specific DH within four weeks of engraftment. In addition, C57BL/6 mice with long-standing, healthy allografts display donor-specific ACAID. However, C57BL/6 mice differ from BALB/c mice in that (1) fewer allodisparate corneal grafts were accepted indefinitely; and (2) rejection in these eyes correlated with sustained donor specific DH. It was concluded that immune privilege is less secure in the eyes of C57BL/6 mice, compared to BALB/c mice. This conclusion is discussed in terms of known polymorphic differences between these mouse strains at genetic loci governing (a) immune responsiveness, (b) alloform of Fas ligand, and (c) bias of immune response toward Th1 or Th2 phenotypes. PMID- 9848223 TI - Modulation of alloimmunity to major histocompatibility complex class I by cotransfer of cytokine genes in vivo. AB - Major histocompatibility complex (MHC) class I antigen is a potent stimulus for alloimmune responses and is the principal immunologic target mediating acute cellular rejection of allografts. Using a method of direct in vivo gene transfer of cDNA encoding donor type MHC class I, we showed in a rat model that recipient muscle could express the transferred MHC class I cDNA, resulting in alloimmunization of the recipient. This was most graphically demonstrated by accelerated rejection of cardiac allografts expressing the same MHC class I as encoded by the immunizing cDNA. We now report the use of the particle-mediated gene transfer via a gene gun (Geneva, Middleton, WI, USA) to transfer MHC class I, as well as cytokine gene expression vectors, into rat skin. Compared to intramuscular injection, gene gun transfer to skin resulted in more efficient immunization. Donor-specific cytotoxic T lymphocyte (CTL) responsiveness and antibody levels increased. Furthermore, coexpression of certain cytokine genes with the MHC class I cDNA modulated the immune response. Specifically, coimmunization with IL-10 cDNA abrogated immunity to allo-MHC class I, while coimmunization with GM-CSF cDNA enhanced it. The influence of expression of these genes in skin was demonstrated by alteration of donor cardiac allograft survival. This model is useful for induction and modulation of alloimmune responses and may be used to develop gene therapy strategies to modify them. PMID- 9848224 TI - Intrathymic administration of B cells induces prolonged survival of fully allogeneic cardiac grafts without prolonged deletion of donor-specific thymocytes. AB - Intrathymic (IT) injection of alloantigen has been shown to induce unresponsiveness to allografts although the exact mechanisms of tolerance induction remains unclear. C57BL/10 (H2b) cardiac allografts were accepted in C3H/He (H2k) mice pretreated with IT inoculation of donor splenocytes (1 x 10(6)) in combination with a depleting anti-CD4 monoclonal antibody 27 days before cardiac transplantation. To investigate which cell types were responsible for tolerance induction by IT injection of alloantigen, resting B (rB) cells or dendritic cells were used as the thymic inoculum instead of whole splenocytes. IT injection of rB cells induced indefinite graft prolongation in all recipients while only 20% of mice that had received IT injection of dendritic cells accepted grafts for over 100 days. In contrast, IT injection of dendritic cells resulted in significant deletion of donor-specific thymocytes whereas rB cells were relatively ineffective. IT deletion is not essential for the induction of tolerance by IT injection of rB cells; nondeletional mechanisms can be involved. PMID- 9848225 TI - Expression and modulation of ICAM-1, TNF-alpha and RANTES in human alveolar macrophages from lung-transplant recipients in vitro. AB - Alveolar macrophages (AMs) play a central role in pulmonary inflammation in response to local stimuli. As a model for investigating anti-inflammatory drugs, we studied the effects of the cyclohexadepsipeptide antibiotic, fusafungine, and that of the glucocorticoid dexamethasone on the expression of ICAM-1, TNF-alpha and RANTES, induced in vitro by rIFN-gamma in human AMs freshly isolated from bronchoalveolar lavage fluid (BAL) obtained in lung-transplanted patients. ICAM-1 antigen expression, induced on AMs after 24 h of culture, was significantly inhibited by fusafungine in a concentration-dependent manner, as measured by flow cytometry analysis using an anti-CD54 monoclonal antibody. TNF-alpha production, but not RANTES release (measured by ELISA), was significantly inhibited. mRNA studies, by means of polymerase chain reaction amplification of complementary deoxyribonucleic acids (RT-PCR), showed no significant modification of mRNA levels, suggesting that fusafungine acts mainly at a post-transcriptional level. In the same conditions, dexamethasone significantly inhibited the release both of TNF-alpha and RANTES by AMs, mainly acting at the mRNA level, but had no effect on ICAM-1 expression. Assessment of the cellular and molecular targets of anti inflammatory drugs in this model of human AM activation should lead to more appropriate treatment of inflammatory process of the respiratory tract. By virtue of its anti-inflammatory effects on alveolar macrophages, combined with its antibacterial properties, fusafungine should prove particularly suitable for local treatment of bacterial infections of the respiratory tract. PMID- 9848226 TI - Genotyping for polymorphisms in interferon-gamma, interleukin-10, transforming growth factor-beta 1 and tumour necrosis factor-alpha genes: a technical report. AB - Polymorphic variants of cytokine genes are associated with acute and chronic transplant rejection. In this technical report, the methods currently used in our centre to genotype individuals for interferon-gamma, interleukin-10, transforming growth factor-beta 1 and tumour necrosis factor-alpha are described in detail. The DNA sequences of primers and probes, and conditions for polymerase chain reactions are given, and the allele and genotype frequencies in our control populations are summarized. PMID- 9848227 TI - Inter- and intraspecific chromosome pattern variation in the yeast genus Kluyveromyces. AB - The analysis of the electrophoretic chromosome patterns of the species of the genus Kluyveromyces, reveals a high polymorphism in size, number and intensity of bands. Different sets of electrophoresis running conditions were used to establish species-specific patterns and also to detect intraspecific variation. According to their karyotypes, the species of this genus can be divided into two major groups. The first group includes the species K. africanus, K. bacillisporus, K. delphensis, K. lodderae, K. phaffi, K. polysporus and K. yarrowii, composing the so-called 'Saccharomyces cerevisiae-like' group, because their karyotypes resemble that of the species S. cerevisiae. The second group comprises the species K. aestuarii, K. blattae, K. dobzhanskii, K. lactis, K. marxianus, K. thermotolerans, K. waltii and K. wickerhamii, whose chromosomal patterns exhibit common characteristics very different to those of the species included in the 'S. cerevisiae-like' group. This division is concordant with the position of these species in previous phylogenetic reconstructions. Additionally, the intraspecific analysis of the chromosome patterns show a rich polymorphism in the heterogeneous species K. dobzhanskii, K. lactis, and K. marxianus, which is in concordance with the variability observed with other phenotypic or genetic markers. On the contrary, K. thermotolerans exhibits a homogeneous karyotype indicative of a very low level of chromosomal polymorphism, which is congruent with the reduced variability found in this species with other molecular markers. PMID- 9848228 TI - Monovalent cation fluxes and physiological changes of Debaryomyces hansenii grown at high concentrations of KCl and NaCl. AB - Debaryomyces hansenii showed an increased growth in the presence of either 1 M, KCl or 1 M NaCl and a low acidification of the medium, higher for the cells grown in the presence of NaCl. These cells accumulated high concentrations of the cations, and showed a very fast capacity to exchange either Na+ or K+ for the opposite cation. They showed a rapid uptake of 86Rb+ and 22Na+. 86Rb+ transport was saturable, with K(m) and Vmax values higher for cells grown in 1 M NaCl. 22Na+ uptake showed a diffusion component, also higher for the cells grown with NaCl. Changes depended on growth conditions, and not on further incubation, which changed the internal ion concentration. K+ stimulated proton pumping produced a rapid extrusion of protons, and also a decrease of the membrane potential. Cells grown in 1 M KCl showed a higher fermentation rate, but significantly lower respiratory capacity. ATP levels were higher in cells grown in the presence of NaCl; upon incubation with glucose, those grown in the presence of KCl reached values similar to the ones grown in the presence of NaCl. In both, the addition of KCl produced a transient decrease of the ATP levels. As to ion transport mechanisms, D. hansenii appears to have (a) an ATPase functioning as a proton pump, generating a membrane potential difference which drives K+ through a uniporter; (b) a K+/H+ exchange system; and (c) a rapid cation/cation exchange system. Most interesting is that cells grown in different ionic environments change their studied capacities, which are not dependent on the cation content, but on differences in their genetic expression during growth. PMID- 9848229 TI - Cloning and characterization of the peroxisomal acyl CoA oxidase ACO3 gene from the alkane-utilizing yeast Yarrowia lipolytica. AB - The ACO3 gene, which encodes one of the acyl-CoA oxidase isoenzymes, was isolated from the alkane-utilizing yeast Yarrowia lipolytica as a 10 kb genomic fragment. It was sequenced and found to encode a 701-amino acid protein very similar to other ACOs, 67.5% identical to Y. lipolytica Aco1p and about 40% identical to S. cerevisiae Pox1p. Haploid strains with a disrupted allele were able to grow on fatty acids. The levels of acyl-CoA oxidase activity in the ACO3 deleted strain, in an ACO1 deleted strain and in the wild-type strain, suggested that ACO3 encodes a short chain acyl-CoA oxidase isoenzyme. This narrow substrate spectrum was confirmed by expression of Aco3p in E. coli. PMID- 9848230 TI - Cloning and characterization of an n-alkane-inducible cytochrome P450 gene essential for n-decane assimilation by Yarrowia lipolytica. AB - A gene encoding cytochrome P450 involved in n-alkane utilization was cloned from an n-alkane assimilating yeast, Yarrowia lipolytica CX161-1B. The RT-PCR was performed on the mRNA prepared from the cells grown on n-alkane as a template using degenerated PCR primers designed for the conserved amino acid sequences of the CYP52 family. The RT-PCR amplified fragment was then used as a probe to isolate genes coding for P450 of the CYP52 family from the genomic DNA library of the strain CX161-1B. The nucleotide sequence of one of the positive clones was determined. An open reading frame which had the same nucleotide sequence as the RT-PCR-amplified fragment was identified. It was of 523 amino acid residues, 60.2 kDa in molecular mass, and had 30-45% sequence identity with the other members of the CYP52 family of Candida species so far analysed. The expression of the P450 gene that was named as YlALK1 was induced by n-tetradecane and repressed by glycerol. A YlALK1 gene disruptant did not grow well on n-decane, but grew on longer-chain n-alkanes such as hexadecane as a sole carbon source. Introduction of YlALK1 on a plasmid to the disruptant restored the decane assimilation. These results suggest that the YlALK1 gene product is the major P450A1k to metabolize short-chain n-alkanes such as decane and dodecane in Y. lipolytica. PMID- 9848231 TI - Isolation and sequence analysis of the orotidine-5'-phosphate decarboxylase gene (URA3) of Candida utilis. Comparison with the OMP decarboxylase gene family. AB - The URA3 gene of Candida utilis encoding orotidine-5'-phosphate decarboxylase enzyme was isolated by complementation in Escherichia coli pyrF mutation. The deduced amino-acid sequence is highly similar to that of the Ura3 proteins from other yeast and fungal species. An extensive analysis of the family of orotidine 5'-phosphate decarboxylase is shown. The URA3 gene of C. utilis was able to complement functionally the ura3 mutation of Saccharomyces cerevisiae. PMID- 9848232 TI - Vectors for transcription factor cloning and target site identification by means of genetic selection in yeast. AB - We describe the construction of a number of vectors that can be used in yeast genetic selection systems for cloning of transcription factors or other DNA binding proteins and for identification of the target sites recognized by transcription factors. For transcription factor cloning we have designed an integration vector with two HIS3 reporter gene cassettes to stably integrate reporter gene constructs at the non-essential yeast PDC6 locus. This set of plasmids was tested in a one-hybrid assay with the rice transcription factor Oshox1, a member of the class of homeodomain leucine zipper proteins. A hybrid protein of Oshox1 and the Gal4 transcriptional activation domain was shown to specifically activate a reporter gene construct with upstream Oshox1 binding sites, which had been integrated at the PDC6 locus using the described vector system. Target site identification by genetic selection in yeast employs a transcriptional activator construct and a library of genomic or random DNA fragments upstream of a reporter gene. We have constructed two variants of a bacteriophage lambda vector which facilitates the construction of the required reporter gene library because of high cloning efficiency and easy conversion into a yeast/Escherichia coli shuttle vector library by Cre-loxP-mediated automatic subcloning. Tests with Oxhox1 as transcriptional activator demonstrated the usefulness of the deprived reporter gene vector. PMID- 9848233 TI - Quantitative analysis of yeast gene function using competition experiments in continuous culture. AB - One possible route to the evaluation of gene function is a quantitative approach based on the concepts of metabolic control analysis (MCA). An important first step in such an analysis is to determine the effect of deleting individual genes on the growth rate (or fitness) of S. cerevisiae. Since the specific growth-rate effects of most genes are likely to be small, we employed competition experiments in chemostat culture to measure the proportion of deletion mutants relative to that of a standard strain by using a quantitative PCR method. In this paper, we show that both densitometry and GeneScan analysis can be used with similar accuracy and reproducibility to determine the proportions of (at least) two strains simultaneously, in the range 10-90% of the total cell population. Furthermore, we report on a model competition experiment between two diploid nuclear petite mutants, homozygous for deletions in the cox5a or pet191 genes, and the standard strain (ho::kanMX4/ho::kanMX4) in chemostat cultures under six different physiological conditions. The results indicate that competition experiments is continuous culture are a suitable method to distinguish quantitatively between deletion mutants that qualitatively exhibit the same phenotype. PMID- 9848234 TI - Current awareness on yeast. PMID- 9848235 TI - [Social work with the elderly--evaluation, crisis and perspectives]. PMID- 9848236 TI - [Repositioning social work with the elderly? Effect of competitive elements and new guidance models on the profile of a profession]. AB - The article discusses changes in the context within which the social professions are exercised. The chances for a refinancing of social work and indeed its very locations change in line with the transition to new steering elements in the social body and the new implementation of competitive elements in the realm of public law. With the economising of the social sphere social work is questioned under economic aspects. Simultaneously it gains normative significance. This is true with regard to standards for the quality of services and institutions, and further in the context of the discussion concerning the revitalisation of local social policy for the elderly. Precisely the economising of the social sphere raises the question whether the municipalities may be excused from their responsibility to form, preserve and support the obligations of social work with the elderly. This article denies this. PMID- 9848237 TI - [Social work with the elderly--current orientation and strategies]. AB - Social work for the elderly in a demographical ageing society is not justified per se, but has to prove its utility under the altering social conditions of the changing age-structure. In the field of practice, this has led to an increasing need for professional orientation and to searching for profiles of performance with greater attraction. As manifold as the target groups are the spectra of social work for the elderly is also manifold in its content and methods. Nevertheless, we are able to recognize some general orientations and strategies, which are conceptual adaptions from social work in general, cultural work and educational work--combined with fundamental supplies of social gerontological knowledge. In this sense, social work, for the elderly--in its most advanced realms--should be integrated into an ageing-policy concept, which is of relevance for the society as a whole. PMID- 9848238 TI - [Ambulatory nursing-care related social work]. AB - From a gerontological point of view, social work which accompanies care processes is absolutely necessary. The social production of care depends crucially on an appropriate attention toward the fitting between informal and formal help systems. Social work can support these fitting processes both on a firm level and on a neutral level. Some specific attributes of social work with elder care dependant living in their private homes are discussed, such as threatening, difficult or ashaming aspects, as well as the necessity of all involved persons agreeing in the developed help-planning process. Practice research and Social science perspectives are discussed as appropriate reference system for life-world orientated social work with informal caregiving arrangements. Concerning the support of informal caregivers as an important example of ambulant social work with elder people, some dimensions of theoretical and practical professionality are proposed. PMID- 9848239 TI - [Personal biography in the aging process and attitude to personal biography in social work with the elderly]. AB - In old age, individualization processes also simultaneously effect the opportunity and necessity to shape one's life. Societal orientation dissipates, and the certainties of earlier times disappear. Plurality and option appear as an increasing twofold challenge to coping with life. These ambiguities characterize modern old age. Or in other words: the biographization of old age has begun. Against this background, the orientation to biography in social work with the elderly is redefined as the reflective handling of the various selfthematizations, which arise, in order to again gain security in unclear circumstances and to reposition oneself. Implied in this, too, is that the orientation to biography requires an scientific foundation and that the naive treatment of narrative life-history in the past is to be overcome. PMID- 9848240 TI - [Basic functions and specialty standards of support management]. AB - Three strategies are followed in handling the key issue within the field of action of care an services for the elderly: securing the material needs of those requiring care, creating appropriate care structures, and providing an effective counselling back-up to alleviate interface problems. Those latter, process oriented aids, have been the subject of discussion and further development under the key term of case management for a good decade. The aim in this context is to prevent the elderly from being unnecessarily moved into residential care, to stop the revolving door syndrome, to ensure that older people can stay in their homes, to support family members acting as care providers, to enhance the quality of services and those to ease the strain on fund providers. In the context of case management, these diverse tasks are performed by three basic substantive functions: the gatekeeper, broker, and advocacy functions. Each of these three functions characterizes the main variants of case management with a different weighting (care management, community care, managed care and case management as a counselling approach with a particular focus on individual help). Although these variants may improve the handling of the key issue in the field of action of care, they nevertheless remain too limited in scope. The integration of a fourth basic function, the support function, leads to a modification and extension of case management and characterises the concept of support management which is better able to fulfill the tasks and standards in the field of action of the elderly population in need of care: interruption of the dynamics leading to residential care, grater transparency through the maze of service provision, the right of self-determination of those requiring assistance in the planning and implementation of care, and quality assurance of care arrangements; it also serves as a contribution to an improvement of care structures. PMID- 9848241 TI - [Social work in geriatric clinics--between autonomy and functional organization]. AB - Geriatric therapy is defined through interdisciplinarity, teamwork, and multiprofessionality. Besides questions of medical therapy, psychosocial dimensions of elderly are highly relevant. Social work is mostly described as an important profession in the process of geriatric therapy. This work refers to the presentation and discussion of the results of an interdisciplinary study to the role of social work in German geriatric hospitals. It shows that the role of social work is determined by the various perceptions of the professions. Another influence on it is by the structures of the hospital. Social workers defines their activities through the relationship to the client; the other professions recognize the tasks of social work in a very diffusely way, mostly reduced in connection with the management of dismission of the patient from hospital. The professional education of social workers is not well known by the nurses and doctors. The fact that social workers are mostly acting under the responsibility of doctors, leads to a definition of problems and aims from a medical point of view. In many of the hospitals there are distinct problems of communication and cooperation between the professions. If the profession of social workers wants to contribute to the reduction of these impediments of communication and cooperation, they must actively and constructively communicate with the other professions and become more clear about their own professional way of acting. PMID- 9848242 TI - [Profile of social work in inpatient elder care]. AB - New ideas in the support of the elderly such as self determination of residents on the one hand, normalization, individualization and opening of geriatric care centers on the other, led to an increasing importance of social work in homes for the elderly. Full quality-management and basic data are instruments for forming a professional profile. On the other hand there is a lack of empirical studies on this topic. This paper presents the results of an activity analysis of 16 persons employed in the social services in homes for the elderly of one non profit organisation in Baden-Wurttemberg. The main work in direct help for the residents consists in the realization and organisation of social activities and in offers of groupwork. This involves measures to structure the day and to promote social contacts. Besides social legal advice, the psycho-social advice for residents and their relatives in order to help them to master critical events and master changes in behaviour caused by gerontopsychiatric illness is of special importance. In an indirect way social work concentrates on the internal organisation of coordinating managed care and social support in a multiprofessional team and other service sectors of the institution. The opening of homes and their integration within the community is achieved by information and public relations work, for example by the cooperation with local clubs, external services and help, and last but not least, by recruiting and advising volunteers. PMID- 9848243 TI - [Possibilities and limits of geriatric assessment in general practice]. AB - Geriatric assessment is a means to collect health and functional data of an individual older person in a standard way. It has been developed on the secondary care level in the specialties of geriatric and rehabilitative medicine. Transferring geriatric assessment into primary care may improve health outcomes of older people, especially as it facilitates preventive diagnostic and therapeutic intervention. Yet, it is not possible to use geriatric assessment, as it is carried out in secondary care, under primary care conditions. Several adaptations will be necessary. An assessment instrument will have to be developed which takes epidemiological features of the older population into account. It will have to be tailored to the specific conditions of health delivery in primary care. Moreover, to achieve effectiveness and acceptance by health care providers and users, we see the necessity of creating an algorithmic assessment instrument which allows the use in different health situations with different levels of diagnostic detail. A primary care assessment for older people then would need to be evaluated according to its effectiveness of improving health outcomes. Potential negative effects on health beliefs may be worth considering. The final task lies in the implementation of a primary care assessment instrument into our existing health delivery format. PMID- 9848244 TI - [Family care of patients with dementia: differential significance of specific stress dimensions for the well-being of caregivers and the stability of the home nursing situation]. AB - In Germany more than 60% of those elderly who suffer from dementia are being cared for by family members within the community. The objective conditions are perceived as extremely burdensome by many caregivers and can seriously affect their psychological and physical well-being. The aim of this study was to discrimininatively analyze the meaning of various dimensions of burden for the carers' well-being and the stability of the caregiving situation. Thus, an explicit distinction is made between caregiver burden which directly refers to the situation and the more general concept of well-being which is potentially, but not necessarily, affected by the caregiving situation. Caregivers' life satisfaction, self-esteem, and their subjective perception of the stability of the caregiving situation (subjective probability of patient's institutionalization within the next 12 months) were regarded as indicators for potential outcomes of caregiving. Regression analysis revealed that to the caregiver's life satisfaction perceived role conflicts are most significant. For the caregiver's self-esteem patient's objective cognitive impairment is decisive. Here the direction of the relation is remarkable: The more severe the patient's impairment, the higher the caregiver's self-esteem. Regarding the stability of the caregiving situation the perceived inadequacy of caregiving resources proves to be of prime importance. Results suggest that a differentiated examination of the complex relations between specific dimensions of burden and potential caregiving outcomes is an adaquate approach to a deeper understanding of the consequences of family caregiving. PMID- 9848246 TI - [Video-assisted interventions in the thorax and abdomen]. PMID- 9848245 TI - [Principles of attachment theory in subjective life satisfaction and individual orientation to the future in advanced adulthood]. AB - Attachment representation was assessed in 48 elderly people with an average age of 69 years. The adult attachment interview was used plus an additional question about wishes for the future. Coherent correlations with subjective life satisfaction and individual future perspective were demonstrated. Elderly participants with secure attachment representation were more satisfied, they were also less anxious, and more often positive about their future. If their adult children had secure attachment representation, they were reported by their elderly parents to be socially, emotionally, and materially more supportive. Attachment representation as a biographical variable is the result of developmental processes. Our findings show qualitative differences in attachment representation with clear consequences for lifestyles in advanced age. Perception and interpretation of environment, and the ability to integrate negative emotions into positive life perspectives is mainly a matter of social-emotional experiences with significant attachment figures. PMID- 9848247 TI - [Minimally invasive interventions in heart surgery]. AB - Recently a number of minimally invasive surgical techniques have been developed in order to reduce surgical trauma especially to avoid median sternotomy and cardio-pulmonary bypass (CPB). In March 1996 we started successfully a clinical trial with the Port Access technique at our institution for the first time in Europe for the treatment of coronary artery single vessel disease. In addition mitral valve disease, aortic valve disease and ASD were treated successfully with minimally invasive surgical techniques. We developed a new minimally invasive surgical technique (Dresden technique) for the treatment of coronary artery multi vessel disease at our institution. Besides we used several minimally invasive surgical techniques without CPB. Our results indicate that minimally invasive surgical techniques routinely used will decrease the morbidity and time of convalescence after cardiac surgery. These techniques can be applied for a variety of cardiac diseases. PMID- 9848248 TI - [Thoracoscopic diagnosis and therapy of lung metastases]. AB - INTRODUCTION: The purpose of this study was to evaluate the impact of thoracoscopy on staging and therapy of peripheral pulmonary nodules in patients with a cancer history. PATIENTS AND METHODS: Videothoracoscopy (VATS) was performed in patients presenting peripheral pulmonary nodules (< 3cm) in CT-scan. 65 patients (63%) presented less than 3 nodules and 39 patients (37%) had multiple lesions in the lungs. History revealed a primary gastro-intestinal cancer in 35 patients (34%), a sarcoma in 26 patients (25%), breast cancer in 13 patients (13%) and miscellaneous primary cancer in 22 patients (21%). VATS was performed under general anesthesia using a standard equipment (Olympus). Double lumen endotracheal intubation was carried out. Thoracoscopic pulmonary resection was accomplished with endoscopic stapler (Autosuture Multi-Fire Endo GIA 30). The specimens were removed in a retrieval bag and a tube was inserted into the thoracic cavity. RESULTS: In 24 patients (23%) conversion to thoracotomy was performed, because of adhesions (n = 12), technical reasons (n = 8), no tumor detectable (n = 4). Thoracoscopic wedge resection for coin lesions was performed in 61 of the 80 patients (76%). Three patients underwent decortication and in 16 patients biopsy was sufficient for therapeutical considerations. Additional informations in comparison to conventional diagnostic were found in 39 patients (49%). The treatment regimen altered in 32 patients (40%). CONCLUSIONS: In this study VATS proved to be a sensitive technique for staging of pulmonary coin lesions. Additional informations were achieved in 49% and therapeutic strategy was changed in 40% of the patients due to the distant spread of malignancy or detection of benign lesions obtained by thoracoscopic staging. PMID- 9848249 TI - [VATS (video-assisted thoracoscopic surgery): possibilities and limits of surgical therapy of malignant lung diseases]. AB - VATS is able to perform by minimal access all operations known in general thoracic surgery. Mortality (0.07-1.9%) is low and complications (4.3-14.2%) are rare, as published in four studies in the last two years. Especially long-term results of VATS in malignant disease are not clear. Indications and limitations for VATS in bronchial carcinoma and lung metastases are discussed. Surgical treatment in peripheral non-small cell lung cancer has best results after lobectomy. Limited resection decreases the oncological result (5-year-survival rate) by 10 to 18% compared to lobectomy. Conservative treatment with radiotherapy in stage I carcinomas in patients who are not eligible for open lobectomy because of restricted lung function or other non oncological reasons has a bad prognosis (0-12% 5-y-survival). In these cases VATS resection of bronchial carcinoma should by considered. Detection of all lung metastases by preoperative radiological study is only in 56-75% possible. Between 10/93-2/98 we performed 52 thoracotomies for lung metastases at our surgical department. In eight cases bronchial carcinoma was confirmed by histological examination, nine patients with metastases of malignant germ-cell-tumors were excluded from the study. CT-scan of the lung was performed preoperatively. In 43% of the cases (n = 35) additional lesions were detected intraoperatively by palpation of the lungs. Histopathologic examination revealed benign lesions in 9% and malignant in 34%. The result of lung metastases resection is highly dependent on the completness of extirpation. We therefore conclude that lung metastases should be removed by open thoracotomy. PMID- 9848250 TI - [Endoscopic interventions in the tracheobronchial system]. AB - Therapeutic bronchoscopy is widely used in the treatment of airway stenosis, fistula, and pulmonary bleeding. Recanalisation of tracheobronchial stenoses can be treated by Nd: YAG-laser or stent implantation. While the use of laser irradiation is limited to intraluminal obstruction, stenting is also effective in stenoses due to extraluminal compression. Stent implantation has also become the method of choice in the treatment of tracheobronchial fistulas. A more specific strategy in the treatment of malignant lesions is photodynamic therapy. By use of photosensitizers and subsequent irradiation tumor cells can be destroyed selectively. Gene therapy represents a promising new approach in tumor therapy. After retrovirus-mediated gene transfer of a suppressor gene an increase of tumor cell apoptosis was observed. Altogether interventional pneumology has experienced substantial enlargement of therapeutic possibilities. PMID- 9848251 TI - [Treatment with stents in the upper gastrointestinal tract]. AB - A wide range of therapeutic methods for treatment of malignant and benign stenoses of the upper gastrointestinal tract is at one's disposal with esophageal mechanical dilatation, laser therapy, insertion of plastic tubes and stent implantation. It has become routine to insert flexible metal stents in the treatment of esophageal carcinoma and tracheoesophageal fistula. Case reports exist describing the use of flexible metal stents in complicated benign esophageal stenoses of stomach and duodenum. Until January 1997 we implanted 18 stents (7 Z-stents[Wilson and Cook], 4 covered and 4 uncovered Ultraflex stents [Boston Scientific], 3 Endocoil stents [Instent]) in 15 patients. We treated patients with esophageal carcinoma, recurrent stenoses after gastrectomy and tracheoesophageal fistula. They were followed by questionnaires. Four days after stent implantation 50% of the patients were able to consume normal solid food, the other half semisolids. The main problem was the stent migration (28%). The decisive factors for stent migration were the quality of surface of the stent and its position in the gastroesophageal junction. The stents which were dislocated in the stomach could easily be extracted endoscopically. The endoscopic extraction of a dislocated Z-stent in the small bowel was impossible. The high rate of stent dislocation (20-30%) in distal stent position (gastroesophageal junction) is not to be tolerated with respect to the purchase price. PMID- 9848252 TI - [Combination of diathermy and argon plasma coagulation in treatment of cicatricial esophageal stenoses]. AB - Scar stenoses of the esophagus are caused by peptic reflux or following surgery of the upper gastrointestinal tract. Short scar cicatricial stenoses are difficult to treat and have a high recurrence rate. Pneumatic dilatations show a short term success. We treated 5 patients with a short scar esophageal stenosis after esophagogastrostomy (n = 3) and after esophagojejunostomy (n = 2). The main symptom was dysphagia. X-ray shows the length of the stenosis (max. 1.5 cm). The stenosis was cutted by diathermy in a starshape. In all 5 cases it was possible to achieve the wide lumen. There were no complications. The combination of diathermy and argon-plasma-coagulation is a safe method, rare of complications in the treatment of short scar stenosis of the esophagus. PMID- 9848253 TI - [Laparoscopic anti-reflux surgery--report of experiences in Austria]. AB - In Austria neither the open nor the laparoscopic fundoplication can be described as a routine operation. The number of laparoscopic operations is increasing year by year, the indication for surgery is more determined by radiological than by functional diagnostic tools. In our own patients the complication rate in 196 laparoscopic operations is 6%, regarding symptoms of reflux and dysphagia. Dysphagia was found only in cases with 360 degrees fundoplication, whereas recurrent reflux occurred in 270 degrees hemifundoplications (Toupet). The mortality rate following laparoscopic fundoplications was zero, morbidity below 10%. PMID- 9848254 TI - [Laparoscopic anti-reflux surgery--report of experiences from Germany]. AB - This is a report on a questionnaire in Germany reflecting the activity in antireflux surgery, both in open and laparoscopic modifications in the time period of 1990 through 1995. It serves as an overview of the acceptance of diagnostic workup, indication, applied techniques, and different antireflux procedures. In a total of 104 representative hospitals, 2,036 patients were operated during this time. Almost 80% of the hospitals provide antireflux surgery in the open technique and only 1/3 of the hospitals have experience in the laparoscopic technique. There is a total rise in antireflux surgery during the last 5 years, since the number of laparoscopic antireflux operations rises constantly with a total amount of open operations of about 250 cases per year. In open surgery the most favourite technique is the Nissen-Rossetti procedure, while in laparoscopic technique the choice for the original Nissen, the Nissen Rossetti, or the floppy Nissen technique is divided in almost equal parts. PMID- 9848255 TI - [Reflux disease: "laparoscopic therapy"]. AB - Laparoscopic surgery offers the patient the advantages of minimally-invasive surgery for the treatment of gastroesophageal reflux disease. Our experience with 46 laparoscopic fundoplications is favorable. The indications for surgery for patients with esophageal reflux disease remain unchanged and should not be extended. However, the good results should facilitate the decision for surgery. Several comparative studies show the safe application of laparoscopic techniques in the surgical treatment of gastroesophageal reflux disease. PMID- 9848256 TI - [The Janeway laparoscopic gastrostomy in palliative surgery]. AB - For the creation of a gastrostomy percutaneous endoscopic technique should be the method of first choice. If impossible a laparoscopic procedure should be preferred to a conventional one. Laparoscopic Janeway-gastrostomy might have some advantages compared to Kader-Stamm-Fistulas. We present the results with this method which we have used since 1993 in 12 patients. The postoperative course of these patients with mainly malignant diseases of ENT (n = 4), esophagus (n = 3) and thyroid gland (n = 3) as well as two non-malignant underlying diseases is presented and discussed in the light of recent publications. The Janeway gastrostomy is an effective, easily feasable and safe procedure that ensures a good palliation also due to easy handling and care. PMID- 9848257 TI - [Laparoscopic rectum surgery in carcinoma]. AB - Rectal cancer requires a special training in laparoscopic colorectal surgery because of its high requests in managing the operation. From August 1993 to March 1997, we performed 61 laparoscopic operations for rectal cancer in our hospital, 53 patients underwent a continence preserving resection and 8 patients a laparoscopic assisted abdominoperineal rectal exstirpation. 50 cases were curatively operated. In 11 patients only a palliative operation was feasible. The perioperative mortality was zero. Intra- and postoperative complications were seen in 7 cases (11.5%). Late complications were observed in another 6 patients (9.8%). 4 of the palliatively operated patients died in a median follow up of 17.5 months. After curative operation a tumor progression occurred in 6 patients (12%), 3 of them (6%) died within the follow up period. Our study demonstrates the feasibility of laparoscopic rectal cancer operations in any localization and with the same oncologic results as achieved with conventional approach. PMID- 9848258 TI - [Surgical intervention in severe acute pancreatitis--retrospective study of 79 patients of the RWTH Aachen Surgical Clinic]. AB - This is a report on the surgical intervention in 79 patients with acute pancreatitis, who were operated in the Department of Surgery of the University Clinic RWTH Aachen in the period from 1986 to 1993. The main objective was the stratification of pancreatitis according to the Ranson-Score, the analysis of the surgical treatment and the timing of operation depending on the clinical condition. The average Ranson-score was 3.3 (median 3). 56 patients had necroses, which were removed because of the deteriorating clinical condition. In these cases the average Ranson-score was 4.2 (median 4). Seven patients (8.9% of the total number and 12.5% of the patients with necroses of the pancreas) died. This small number is the result of a severity-adapted management in a modern intensive care-unit and the good cooperation with the Department of Internal Medicine. PMID- 9848259 TI - [Esophageal cyst--a rare cause of backache]. AB - Cysts of the esophagus are rare mediastinal tumors presenting a large variety of clinical symptoms. Even with modern methods of examination an exact diagnosis is difficult to obtain and often only possible after histological examination. Complications are perforation, or pulmonary, thoracic or esophageal haemorrhage. First choice of therapy is the exstirpation. We report on a 50 year old man with backache and the initial diagnosis of renal colic. Only the magnetic resonance image showed an isolated mediastinal esophageal cyst as the reason. The cyst was removed by thoracotomy. The operation and treatment were without complications. Symptomatology, diagnostical pitfalls and the therapy are described. PMID- 9848260 TI - [Colon carcinoma after ureterosigmoidostomy--a case report]. AB - Ureterosigmoidostomy was the method of choice for continent urinary diversion in cases of bladder exstrophy. The development of colonic cancer at the ureterocolic junction is one of the major complications in the late course of this procedure. The risk of large bowel carcinoma is increased 150-500-fold compared to the general population. Therefore regular examinations are mandatory. Based on a case report the problems associated with ureterosigmoidostomy are presented. PMID- 9848261 TI - [Bochdalek hernia in Adulthood: case report and overview]. AB - A 34 year-old female patient was admitted to our hospital with left-sided abdominal pain. Physical examination, chest and abdominal X-rays led to the diagnosis of a colonic obstruction. Colonic gastrografin study showed the splenic flexure herniated into the left hemithorax. Bochdalek hernia was diagnosed and emergency operation was performed. Postoperative course was uneventful. PMID- 9848262 TI - [Determination of the current status of antibiotic prophylaxis in surgical interventions. Workshop of 4 and 5 September 1998 in Gravenbruch]. PMID- 9848263 TI - [Recommendations on contents of medical reports in rectal carcinoma: I: Surgical report and pathology findings. Orientating aids in interdisciplinary management]. PMID- 9848264 TI - [Operations of the chest wall and thorax]. PMID- 9848265 TI - Use of sociometric techniques to assess the social status of mainstreamed children with learning difficulties. AB - Research on sociometric data collection and analysis methods is reviewed and implications for assessing the social status of mainstreamed children with learning difficulties are evaluated. Recommendations are made for changing existing procedures to account for factors specific to children with learning difficulties and to classrooms where mainstreaming is occurring. Variations between frequently used sociometric classification systems (which categorize children as popular, rejected, average, neglected, and controversial) are described, and information on their reliability and validity is discussed. Further reliability research with mainstreamed children is recommended, as is the application of theoretical accounts of affiliation in designing sociometric methods. PMID- 9848266 TI - An organizational model of choice: a theoretical analysis differentiating choice, personal control, and self-determination. AB - Individuals experience choice when they select one option from among meaningful alternatives that possess relatively equal attractiveness and some degree of indeterminacy. Choice has been found to influence important psychological and behavioral outcomes. After differentiating among choice, personal control, and self-determination, the author offers a model of choice, with self-determination as the key mechanism regulating how choice influences intrinsic motivation. The model suggests specific types of choice-relevant information that should affect whether choice results in an internal (self-determined) or external (controlled) locus of causality. The individual characteristics of locus of control, self presentation, self-esteem, and Type A personality are suggested as possible moderators of the effects of choice. Finally, the implications of the choice model for organizations and further areas of research are discussed. PMID- 9848267 TI - Mutual attachment, personality, and drug use: pathways from childhood to young adulthood. AB - Young adult drug use stemming from childhood aggression, the parent-child mutual attachment relationship, and the effect of unconventionality were studied. Youngsters and their mothers were interviewed when the former were early adolescents, late adolescents, and young adults. Additional data were collected from the mothers when their youngsters were children. The analysis was conducted on youngsters who had complete data at all 4 points in time. The findings were in accord with the family interactional model; that is, the parent-child mutual attachment relationship affects unconventionality in the youngster, which, in turn, affects young adult drug use. The results indicate that the parent-child mutual attachment relationship does so through (a) the stability of the attachment relationship from childhood to young adulthood, (b) the stability of unconventional personality and behavioral attributes from early adolescence to young adulthood, and (c) the stability of drug use from early adolescence to young adulthood. The findings imply that (a) early intervention with respect to aggression, (b) interventions that focus on strengthening the parent-child bond and conventional behavior, and (c) interventions aimed at early drug use should be most effective in reducing young adult drug use. PMID- 9848268 TI - Estimation of subcellular organelle volume from ultrathin sections through centrioles with a discretized version of the vertical rotator. AB - A method for the fast and efficient estimation of the volume (but not surface area) of subcellular organelles is presented. It consists of a rotator/coaxial section approach based on the Pappus theorem and represents a discretized version of the vertical rotator where, instead of measuring intercept lengths, the points in distance classes are counted. Centrioles serve as a unique reference 'double point' with constant size allowing unbiased cell selection from the whole population with equal probability and without the disector application. The sandwich-like method of sample preparation allows comparison of control and experimental cases with the same errors caused by overlapping and overprojection. Test experiments demonstrated that the vertical discretized rotator was an efficient and precise tool for the estimation of volume and that a few independent sections of unknown thickness were sufficient for the quantification of one experimental point. PMID- 9848269 TI - Registration of serial sections of mouse liver cell nuclei. AB - Image registration of biological tissue is essential for 3D reconstruction, which is important for visualizing and quantifying the 3D relationships between internal structures of an object. The biological role of DNA organization, which is an extremely complex 3D architecture within the cell nucleus, has come into focus since it has become clear that the chromatin structure in itself functions as a regulator of DNA. Thus, 3D reconstruction of cell nuclei based on consecutive series of high-resolution ultrathin slices may provide new information about the chromatin structure and its organizational changes during carcinogenesis. This work focuses mainly on the problem of registering successive serial transmission electron micrographs of ultrathin sections of mouse liver cell nuclei to analyse the 3D chromatin structure. A five-step semiautomatic interactive registration method is proposed. The first two steps of the procedure correct the rotation and translation components by using the phase correlation. The third, fourth and fifth steps correct the global distortion, employing a point mapping method based on different ways of selecting the control points. In step three, the control points were automatically computed by phase correlating corresponding subimages of the reference and sensed image. A semiautomatic method is used in the fourth step to select the control points, i.e. an automated method for computing the centre of mass of manually identified anatomical structures in neighbouring slices. For the sections which could not be properly corrected by the four steps, a final step is introduced, where control points are manually selected in the reference and sensed images. An algorithm is proposed to examine the spatial distribution of selected control points. Four sets of serial sections of mouse liver cell nuclei, each with approximately 100 sections, are registered by the proposed method and also registered manually for the comparison of registration accuracy. Artificial X-Z and Z-Y sections of registered series were visually compared for the smoothness of the nuclear membrane. To quantify the registration accuracy and the extent of registration, the correlation coefficient (C) and the overlap index (Co) were computed over the registered structure of interest. In addition to the visual comparison and the comparison of C and Co, the registered serial sets were compared by 3D GLCM-based texture features in the Z direction. The results demonstrate that the proposed semiautomatic registration technique achieved accurate results comparable to the manual registration. The proposed registration method relies only on the operator for rough pinpointing of cellular structures. Therefore, it should provide better reproducibility, and allow the user to operate the system faster and in a more relaxed manner than in a manual registration. PMID- 9848270 TI - Repetitive cell 'jumps' during hypotonic lysis of erythrocytes observed with a simple flow chamber. AB - This is the first report describing cell movements of a repetitive character during hypotonic lysis of erythrocytes (haemolysis). A new, simply constructed chamber is described for microscopy of freely suspended cells, for example, blood cells, during the inflow of a new medium. Hypotonic haemolysis of individual red blood cells was studied. During the first phase of haemolysis discontinuities were found: the cells made between zero and seven sudden movements or 'jumps', interpreted as caused by an ejection of cytoplasm due to excess intracellular hydraulic pressure and the formation of a hole. After pressure equilibration the hole resealed spontaneously. When, after one or two jumps, the inflow of hypotonic medium was stopped, the haemolytic process was interrupted but continued after restarting the flow. Inhibition of haemoglobin (Hb) release by 80% by external Ficoll did not affect the number of 'jumps'. Since the optical contrast was reduced owing to Hb release after the last jump, less than 20% of the Hb loss can be associated with the jumps. Ejections of faint clouds of Hb were observed mainly in the presence of Ficoll, but only after the last jump. PMID- 9848271 TI - The axonal transmission of herpes simplex virus to epidermal cells: a novel use of the freeze substitution technique applied to explant cultures retained on cover slips. AB - Retaining the ultrastructural arrangement of a mixed-cell culture on a solid support while processing for immunocytochemical study is a technical challenge. We developed a technique to study the axonal transport of the Herpes simplex virus from dorsal root ganglia sensory neurones to epidermal cells. Autologous explants of human foetal dorsal root ganglia and skin were cultured on plastic cover slips. Axon fascicles grew from the ganglia to the epidermal cells and the ganglia were inoculated selectively with virus. The whole preparation, retained on the cover slip, was fixed with formaldehyde 4% (freshly prepared from paraformaldehyde)/glutaraldehyde 0.1%, processed by freeze substitution, and embedded in Lowicryl HM20 resin. The edges of the cover slip in the block were trimmed, allowing clean and complete separation from the resin block, which retained the tissue. The resin block was placed in fresh HM20 and repolymerized. The polymerizing resin bonded strongly to the existing block. After trimming, serial sections were easily obtained and successfully immunolabelled for viral proteins. This is a convenient technique for immunolabelling tissue grown on cover slips in which the preservation of the ultrastructural interactions between different cells is important. It should be adaptable to a number of cell-culture applications and has a number of advantages over other techniques. PMID- 9848272 TI - Modelling DNA damage induced by different energy photons and tritium beta particles. AB - PURPOSE: To model the production of single- and double-strand breaks (ssb and dsb) in DNA by ionizing radiations. To compare the predicted effectiveness of different energy photon radiations and tritium beta-particles. MATERIALS AND METHODS: Modelling is carried out by Monte Carlo and includes consideration of direct energy depositions in DNA molecules, the production of species, their diffusion and interactions with each other and DNA. Computer-generated electron tracks in liquid water are used to model energy deposition and to derive the initial positions of chemical species. Atomistic representation of the DNA in B form with a first hydration shell is used. Photon radiations in the energy range 70keV-1MeV and tritium beta-particles are considered. RESULTS: A tentative increase for dsb yield has been predicted for 70 keV photons and tritium compared with 137Cs. This increase is more pronounced for complex dsb. Double-strand breaks are much more prone compared with ssb to combine with additional strand breaks and base damage, which contributes to break complexity. At least half of DNA breaks are hydroxyl radical mediated. CONCLUSIONS: The developed model makes predictions compatible with features of available experimental data. Break complexity has to be addressed in biophysical modelling when the relative effectiveness of radiations in DNA damage is studied. Obtained data strongly argue against the dominance of direct radiation action in DNA damage in the cellular environment predicted by some theoretical studies. PMID- 9848273 TI - Repair of radiation-induced DNA double-strand breaks in human fibroblasts is consistent with a continuous spectrum of repair probability. AB - PURPOSE: To propose a novel interpretation of DNA double-strand break (dsb) repair based on the distribution of energy micro-deposition. MATERIALS AND METHODS: Double-strand break repair curves were studied either after irradiation at 4 degrees C or at 37 degrees C (low dose rate). Two human fibroblast cell lines were used: a control line, HF19, and an ataxia telangiectasia repair deficient line, AT5BI. Irradiations were made with gamma-rays or alpha-particles (241Am). Repair data were fitted by the variable repair half-time (VRHT) model. Assuming that each dsb has its own inherent repair half-time (IRHT) and that the VRHT is the average of the IRHT at any time during repair, the distribution of the IRHT was calculated. RESULTS: At the end of the irradiation, the distribution was a continuous asymmetric curve with a maximum of dsb having a short IRHT. After 1 h of repair, the curve became bell-shaped. There is a striking similarity between the distribution of dsb repair half-times and that of energy micro deposition described by Goodhead et al. (1993). CONCLUSION: This similarity suggests a possible causal relationship between the energy density deposition and the repair rate or the probability of dsb repair. PMID- 9848274 TI - A highly conservative, cyclically permuted, non-homologous exchange among three unrelated DNA sequences in bleomycin-treated CHO cells. AB - Molecular analysis of a bleomycin-induced rearrangement of the aprt gene in CHO cells revealed that it consisted of a nearly perfect three-way exchange among non homologous sequences, consistent with a mechanism involving cyclically permuted misjoining of the six ends of three double-strand breaks. PMID- 9848275 TI - Persistence of translocations after accidental exposure to ionizing radiation. AB - PURPOSE: To assess the validity of translocations for retrospective biodosimetry using FISH chromosome painting of peripheral lymphocytes in repeated samples of people accidentally exposed to radiation. MATERIALS AND METHODS: Blood samples from five people exposed to mainly whole-body irradiation of gamma-rays from a radiation accident in Estonia in 1994 were taken at 2-4 month intervals. A total of eight samplings were carried out, including one directly after the accident. Fluorescence in situ hybridization with probes for chromosomes 1, 2 and 4 was performed on metaphase preparations from 48 h whole blood cultures; 1500 cells were scored from each individual per time point. RESULTS: Translocations remained relatively stable during 2 years after exposure in all subjects. A noticeable decrease in complete translocation yields and a significant decrease in incomplete translocations were observed in one person. In addition to whole-body exposure, he had also been exposed to partial-body irradiation. Due to the overall persistence of translocations, dose estimates were very similar throughout the 2 year period. A rapid decline in dicentric frequencies was noted during the first year after exposure. CONCLUSIONS: The results suggest that during 2 years the yield of translocations in peripheral lymphocytes remained at a constant level after whole-body exposure. This finding supports the use of translocations for retrospective dosimetry, at least within this relatively short period of follow-up. In the case of partial-body irradiation, however, the elimination of co-existing unstable aberrations reduced the translocation yield over time. Follow-up will be continued in order to determine the stability of translocations over longer times. PMID- 9848276 TI - Comparison of radiation-induced aberration frequencies in chromosomes 1 and 2 of two human donors. AB - PURPOSE: The purpose of this study was to analyse donor and time dependent variations in the frequencies of radiation-induced aberrations in chromosomes 1 and 2. MATERIALS AND METHODS: Human lymphocytes from two donors were irradiated with 1 and 2 Gy of X-rays. Chromosomal aberrations were scored in chromosomes 1 and 2 painted with different fluorochromes and in Giemsa stained cells. Two time displaced experiments were performed with lymphocytes of each donor. RESULTS: In cells of both donors chromosome 1 was generally more frequently involved in translocations than chromosome 2. This result was not always reproducible. Chromosome 2 showed a higher frequency of acentric fragments, especially following a dose of 1 Gy. Again interexperimental variations were observed. No differences between the two chromosomes were seen with regard to other aberration types. Both chromosomes showed less dicentrics and more acentric fragments than proportional to their DNA content. CONCLUSIONS: Chromosomes 1 and 2 show a different sensitivity to ionizing radiation. The difference is dependent on the aberration type and is not always reproducible. This variability could contribute to the difficulty in reaching a consensus regarding the radiosensitivity of individual chromosomes. PMID- 9848277 TI - A non-parametric method of reconstructing single-dose survival curves from multi fraction experiments. AB - PURPOSE: A method of estimating the single-dose curve from designed multifraction experiments is described and applied to three datasets. MATERIALS AND METHODS: The method, which is non-parametric and based on standard statistical regression techniques, can be used for functional endpoints which are either continuous or binary. The datasets are concerned with wound healing on mice, myelopathy in guinea pigs and spermatogenesis in mice. The results are compared with the results from fitting the linear quadratic model. The statistical methods of Bootstrapping and residual plots are illustrated. RESULTS: The method is based in part on an assumed statistical model, however, exact knowledge of the correct statistical model is not necessary to obtain an estimate of the shape of the single-dose survival curve. We find no good evidence from the reconstructed single-dose survival curve of an "induced repair" phenomena at low doses for the wound healing and spermatogenesis experiments. For the myelopathy experiment the data are consistent with the LQ model with a low alpha-beta ratio down to doses of at least 1.5 Gy per fraction. CONCLUSIONS: A robust statistical method of estimating the shape of the single-dose survival curve is demonstrated using standard statistical software on three datasets. PMID- 9848278 TI - Human cell membrane oxidative damage induced by single and fractionated doses of ionizing radiation: a fluorescence spectroscopy study. AB - PURPOSE: To investigate the production and repair of lipid oxidative damage in two human cell lines exposed to acute and fractionated dose of ionizing radiation. Radiation dose was in the range from 0.1 to 44 Gy. MATERIALS AND METHODS: K562 and HL60 human cell lines have been used, 24 and 96 h after seeding. Membrane lipid oxidative damage has been detected by the measurement of the fluorescence decay of 1,6-diphenyl-1,3,5-hexatriene (DPH), its polarization value and the conjugated dienes concentration. The modification of DPH decay has been previously reported to be directly related to the lipid hydroperoxide concentration. RESULTS: A modification of the DPH decay has been observed as a linear function of the logarithm of the radiation dose and only when the irradiation was performed in the presence of oxygen. The amount of the damage is related to the time after the cell medium change. By exposing the cells to fractionated radiation doses for several days (10 cGy day(-1)), the oxidative damage has been found to be cumulative. After a single acute dose, evidence of repair of the lipid oxidative damage was not obtained. CONCLUSIONS: Following a previously developed method, the membrane damage was attributed to the production of hydroperoxide residues in the lipid acyl chains with the consequence of water penetration into the external portion of the bilayer, from the aqueous environment to the position of hydroperoxides. This damage is not repaired. The results obtained by measuring the DPH fluorescence decay have been compared with those obtained using other current optical and biochemical methods. None of these techniques could detect membrane oxidative damage at doses < 10 Gy. Finally, the different sensitivity of 'young' and 'old' cells to the oxidative damage can be related to different cholesterol concentrations. PMID- 9848279 TI - Cholesterol induced variations of membrane dynamics related to the induction of apoptosis in mouse thymocytes. AB - PURPOSE: To evaluate the involvement of cholesterol induced variations of membrane dynamics in mouse thymocyte apoptosis. MATERIALS AND METHODS: Membranes of thymocytes of RK mice were enriched with cholesterol using methyl-beta cyclodextrins as carriers. Spontaneous apoptosis was compared with apoptosis induced either by X-irradiation, by treatment with dexamethasone (DEX), and by phorbol-12-myristate-13-acetate (PMA). Apoptotic cells were quantified by means of flow cytofluorometry. RESULTS: Small amounts of incorporated cholesterol enhance the cellular sensitivity for spontaneous apoptosis whereas larger amounts of incorporated cholesterol protect against spontaneous apoptosis and apoptosis induced by irradiation, DEX, or PMA. CONCLUSIONS: Cholesterol exerts specific rigidity effects on lipid membranes which have been shown to be involved in thymocyte apoptosis. The general effect of higher concentrations of cholesterol protecting against apoptosis hints towards a central protective mechanism. This study believes that either cholesterol paralyses great parts of the cell metabolism or that the apoptotic chain reaction is interrupted at a central point due to protection of membrane lipid regions from oxidative stress. PMID- 9848281 TI - A substance P receptor antagonist (FK 888) modifies gut alterations induced by ionizing radiation. AB - PURPOSE: We previously reported disturbances of ileal substance P (SP) levels and of characteristics of specific receptors after ionizing radiation associated with disorders of intestinal motility. The aim of this study was to investigate the effect of a SP receptor blockade by FK 888 on gut SP levels and contractile properties after rat irradiation. MATERIALS AND METHODS: Rats were exposed to 6 Gy whole-body gamma-irradiation and injected 1 h post-irradiation with FK 888 for 3 days (0.1 mg/kg/day). Plasma and ileal SP concentrations, ileal muscle SP receptor binding and SP-induced contractions in isolated ileum were investigated 3 and 14 days post-irradiation and FK 888 treatment. RESULTS: Irradiation induced an increase of total SP binding site number at day 3 (1.3-fold) and day 14 (1.6 fold). FK 888 had no effect on SP receptor characteristics in irradiated animals. In contrast, FK 888 treatment caused a reduction of endogenous ileal SP level in mucosal (-29%) and muscularis (-40%) layers at day 3 and these decreases were greater at day 14, -88% in mucosal and -61% in muscularis layers. FK 888 treatment decreased efficacy of ileal contraction in both the control and irradiated rat but surprisingly it increased potency at day 3 and decreased it at day 14 in the irradiated rat. CONCLUSIONS: The findings demonstrate that a SP receptor antagonist could be effective on intestine contractility alteration induced several days after ionizing radiation exposure but not at 3 days after irradiation. PMID- 9848282 TI - Definitions of physical and biological low dose. AB - The concept of 'low dose' may be understood to refer to an average frequency of microdosimetric events (charged-particle traversals) that is substantially less than unity in cells or their nuclei. An important consequence is that in this case the probability of any effect on autonomous cells must be proportional to the absorbed dose and independent of dose rate. However, this definition may be unnecessarily restrictive because--especially in the case of low-LET radiation- only a small fraction of events may cause the effects under consideration (e.g. cell lethality). This results in larger 'biological' rather than 'physical' 'small doses'. From a pragmatic viewpoint, one may consider the fact that in the linear-quadratic model deviations from proportionality between effect probability and absorbed dose are attributed to a term that depends on the square of the absorbed dose. This permits the selection of a criterion which establishes as 'small doses' those in which such deviations are less than a chosen value which in the examples given here is 10%. Different applications of this criterion to the inactivation of V 79 hamster cells are considered. PMID- 9848280 TI - IkappaBalpha degradation is not a requirement for the X-ray-induced activation of nuclear factor kappaB in normal rat astrocytes and human brain tumour cells. AB - PURPOSE: To investigate the mechanism of NFkappaB activation by X-rays in normal primary rat astrocytes. MATERIALS AND METHODS: Primary cultures of type I astrocytes generated from the cortex of neonatal rats were exposed to X-rays with and without various kinase inhibitors and a protease inhibitor. The nuclear or cytoplasmic protein extracts were collected at specified times after treatment and analysed for NFkappaB-DNA binding activity and IkappaB protein levels. RESULTS: The NFkappaB-DNA binding activity was induced by X-rays in a dose- and time-dependent manner in the absence of IkappaB protein degradation in astrocytes as well as in the human glioma cell line U-373MG. Whereas a protease inhibitor (calpain inhibitor 1) and a protein kinase C inhibitor (CGP-41251) did not affect X-ray-induced NFkappaB-DNA binding, treatment of astrocytes with the tyrosine kinase inhibitor (erbstatin) completely prevented the increase in NFkappaB activity after irradiation. Erbstatin also reduced the phosphorylation of IkappaBalpha after X-ray exposure. CONCLUSIONS: These results indicate that, in contrast with the more frequently investigated activators of NFkappaB, radiation induced activation of this transcription factor proceeds in the absence of IkappaBalpha degradation and requires tyrosine phosphorylation. PMID- 9848283 TI - Effect of gamma radiation on the foetal haemopoietic system in the mouse. AB - PURPOSE: To study the effect of gamma irradiation, in the late foetal period, on the haemopoietic progenitor cells of the mouse foetus. MATERIALS AND METHODS: Pregnant Swiss mice were exposed to 0.1 to 1.5 Gy of Cobalt-60 gamma radiation on the 17th day of gestation. At 24h (18 day p.c.) and 72 h (20 day p.c.) after exposure the weights of foetal liver and spleen and their mean cellularity were determined. Chromosomal aberrations and micronuclei (MN) were studied at 24 h. Haemopoietic progenitor cell survival at 24 and 72 h post-irradiation was studied by exogenous spleen colony assay. RESULTS: Irradiation caused significant reduction in the weights of foetal liver and spleen, which was more pronounced at 72 h post-irradiation. The mean cellularity as well as the CFU-S8 and CFU-S12 of liver and spleen showed a radiation dose-dependent decrease, but the spleen showed a higher sensitivity than liver to doses below 1.0 Gy. Chromosomal damage in liver increased linearly with dose, while in spleen the increase was linear quadratic. CONCLUSIONS: The results demonstrate the damaging effect of low doses of gamma radiation on the late foetal haemopoietic progenitor cells of the mouse. Induction of cytogenetic damage appears to be a no-threshold effect in the dose range used. PMID- 9848284 TI - EPR and spectrophotometric studies on free radicals (O2.-, Cysa-HB.-) and singlet oxygen (1O2) generated by irradiation of cysteamine substituted hypocrellin B. AB - PURPOSE: The aim of this work was to investigate the photodynamic action of cysteamine substituted hypocrellin B (Cysa-HB) photochemically and survey the effect of oxygen on its reaction mechanism. MATERIALS AND METHODS: The EPR method is used to detect the generation of the semiquinone anion radical of Cysa-HB (Cysa-HB.-), and active oxygen species using DMPO and TEMP as spin traps. DPA bleaching method is used to determine the quantum yield of 1O2 generated from Cysa-HB photosensitization. RESULTS: In anaerobic solution, Cysa-HB.- was predominantly photoproduced via the self-electron transfer between the excited and ground state species. The presence of electron donors significantly promotes the reduced form of Cysa-HB. When oxygen-saturated solutions of Cysa-HB were illuminated with visible light, superoxide anion radical (O2.-) and singlet oxygen (1O2) were formed. The superoxide anion radical was generated by Cysa-HB.- via electron transfer to oxygen, and this process was significantly enhanced by the presence of electron donors. The 1O2 quantum yield is estimated to be 0.71. Moreover, the accumulation of 1O2 will be replaced by that of Cysa-HB.- with the depletion of oxygen. CONCLUSIONS: These observations suggest that the photodynamic action of Cysa-HB may proceed via both a Type I and Type II mechanism, and that a Type II mechanism will transform into a Type I mechanism as oxygen gets depleted. PMID- 9848285 TI - Cost-effective treatment of the infertile couple. AB - OBJECTIVE: To review the published literature on the cost-effective approach to infertility treatment. DESIGN: The literature on the economics and cost effectiveness of infertility treatments was reviewed. Studies related to this topic were identified through MEDLINE. RESULT(S): Few cost-effectiveness studies about infertility treatment have been published. In the absence of tubal blockage and severe male factor, use of IUI and hMG-IUI is more cost-effective than IVF. In vitro fertilization is at least as cost-effective as tubal surgery. Although IVF costs are high, they fall well within the range of other accepted medical treatments and are below the general public's willingness to pay for these treatments. CONCLUSION(S): Cost-effectiveness analysis is an important means of improving quality of care while controlling costs. Further work regarding cost effectiveness of treatments among different diagnostic groups is needed. PMID- 9848286 TI - Strategies to improve insurance coverage for infertility services. AB - Various studies repeatedly have demonstrated that in the United States, only a minority of couples with infertility receive medical treatment and even fewer receive such treatment from specialists. Primarily couples of higher socioeconomic status receive such treatment. It is therefore obvious that a lack of insurance coverage greatly contributes to this country's disproportionately low utilization of fertility services compared with other developed nations. Such low utilization raises significant ethical and economic issues. For example, if the treatment of infertility is to be perceived as the treatment of a disease stage (a notion widely supported in our professional community), then how can we defend the fact that services are principally available only to the affluent? Moreover, by having access to only a fraction of the potential national market for infertility services, the national overhead for these services is distributed over only a fraction of potential buyers, greatly increasing unit costs. In short, obtaining a larger market for our services through better insurance coverage would solve ethical quandaries and, at the same time, reduce fertility costs on a unit level. How, then, can such improved insurance coverage be achieved? A number of possible strategies come to mind. PMID- 9848287 TI - Do attitudes toward disclosure in donor oocyte recipients predict the use of anonymous versus directed donation? AB - OBJECTIVE: To compare the demographic and psychological characteristics of oocyte recipients and to determine whether the issue of disclosure about the use of a donor is a correlate of the decision to use an anonymous or directed donor. DESIGN: Cross-sectional study. SETTING: University teaching hospital. PATIENT(S): Ninety consecutive recipients of donated oocytes (64 of whom used anonymous donors and 26 of whom used directed donors). INTERVENTION(S): Pretreatment psychosocial evaluation. MAIN OUTCOME MEASURE(S): Recipient opinions and attitudes regarding the choice of donor type and disclosure to others as determined through a semistructured interview. RESULT(S): There were no statistically significant differences with regard to demographic characteristics between recipients who used anonymous and directed donors. There were statistically significant differences between the groups with regard to the issue of disclosure. Recipients who used directed donors were more likely to have told others about using an oocyte donor and were more likely to indicate that they intended to inform the child about the nature of his or her conception. CONCLUSION(S): Oocyte recipients who use known donors differ significantly from those who use anonymous donors with regard to the issue of disclosure to others. Further studies are needed to determine the causal direction of this relation. PMID- 9848288 TI - Effects of ovarian source, patient age, and menstrual cycle phase on in vitro maturation of immature human oocytes. AB - OBJECTIVE: To determine the efficiency of in vitro maturation, expressed by nuclear maturation, of oocytes aspirated during gynecologic surgeries or collected from excised ovaries. To assess the effect of patient age and cycle phase at collection on the oocyte's ability to mature in vitro. To examine the time course of oocyte maturation in vitro. DESIGN: Nuclear maturation based on patient criteria compared. SETTING: University-based IVF program and research center. PATIENT(S): Consented patients undergoing gynecologic surgeries or patients undergoing oophorectomy. INTERVENTION(S): Oocytes were maintained in culture for 48 hours and evaluated for maturation. MAIN OUTCOME MEASURE(S): Nuclear maturation evaluated as germinal vesicle breakdown (GVBD) or progression to the metaphase II (MII) stage. RESULT(S): A significantly higher percentage of oocytes collected during the follicular phase of the menstrual cycle underwent GVBD than did oocytes collected during the luteal phase (60% versus 48%, respectively). The percentage of oocytes reaching the MII stage, from these two groups, was not different. No statistically significant differences in maturation were observed in oocytes from different ovarian sources or from patients >40 or <40 years of age. CONCLUSION(S): These data suggest that oocytes collected during the follicular phase are more likely to undergo GVBD than oocytes collected during the luteal phase. In this study, ovarian source, age, or cycle phase did not influence the final meiotic maturation of oocytes to metaphase II. PMID- 9848289 TI - Factors affecting the success of human blastocyst development and pregnancy following in vitro fertilization and embryo transfer. AB - OBJECTIVE: To determine the factors affecting blastocyst development and pregnancy after IVF and ET. DESIGN: Retrospective analysis of data arising from a clinical trial. SETTING: Private in vitro fertilization clinic. PATIENT(S): Fifty six patients aged < or = 40 years, undergoing IVF procedures for infertility, recruited specifically for blastocyst transfer. INTERVENTION(S): All zygotes were cultured to days 5 or 6 after insemination, and one to four of the most advanced blastocysts were transferred to the patient's uterus. MAIN OUTCOME MEASURE(S): Development of zygotes to blastocysts in vitro and pregnancy and implantation rates after ET. RESULT(S): Fifty-one percent of all zygotes developed to blastocysts. Significant positive correlation between the number of blastocysts formed was observed with the number of oocytes, pronuclear zygotes, and eight cell embryos formed. There was a negative correlation with male factor infertility. By day 5 or 6, 93% of the patients had at least one blastocysts, and the clinical pregnancy rate per transfer was 43% and the implantation per embryo transferred was 25%. No other clinical factor significantly affected the number of blastocysts formed, pregnancy rate, or implantation rate. CONCLUSION(S): The numbers of oocytes, zygotes, and normally developing embryos in culture significantly affects the production of blastocysts in vitro. Male infertility significantly reduces blastocyst production. The number and the quality of the blastocysts transferred significantly influences clinical pregnancy rate. PMID- 9848290 TI - One thousand initiated cycles of in vitro fertilization in women > or = 40 years of age. AB - OBJECTIVE: To evaluate the results of IVF in women > or = 40 years of age using their own oocytes. DESIGN: Retrospective study. SETTING: Wolfson and Royal Masonic in vitro fertilization units, London, United Kingdom. PATIENT(S): A total of 1,087 IVF cycles were started in women > or = 40 years of age. INTERVENTION(S): Medical records of patient outcomes were reviewed. MAIN OUTCOME MEASURE(S): Clinical pregnancy, miscarriage, and delivery rates. RESULT(S): Of the 1,087 cycles started in 471 women > or = 40 years of age, 842 reached oocyte retrieval (77.5%) and 702 had embryos transferred (64.6%). The pregnancy rate (PR) was significantly lower in women > or = 40 years of age than in a control group of women <40 years of age (11.3% versus 28.2%). It decreased sharply in women >42 years of age, and no women >45 years of age had a child. Women > or = 40 years of age were more likely to miscarry (27% versus 12.7%). When only one embryo was available for transfer, the PR was 3.3%. When >2 embryos were available for transfer, the PR was similar whether 2 or 3 embryos were replaced. No triplet pregnancy occurred. Women > or = 40 years of age achieved a cumulative PR of 30% after three cycles with a cumulative "take home baby" rate of 21%. CONCLUSION(S): In vitro fertilization is a reasonable treatment for women <45 years of age using their own gametes. Those with a "good response" in their first attempt may be encouraged to complete three cycles with an acceptable chance of conception. PMID- 9848291 TI - Effect of salpingectomy on ovarian response to superovulation in an in vitro fertilization-embryo transfer program. AB - OBJECTIVE: To investigate the effect of salpingectomy on the response of each ovary in patients undergoing an IVF-ET treatment cycle and to compare the results with those of patients who had not had surgery and were undergoing IVF-ET during the same period. DESIGN: A prospective study. SETTING: Tertiary referral academic IVF unit. PATIENT(S): Twenty-nine ET cycles were evaluated in 29 patients who previously had undergone unilateral salpingectomy because of ectopic pregnancy (study group). Seventy-three patients with unexplained or male factor infertility served as controls. INTERVENTION(S): Ovulation induction and IVF-ET. MAIN OUTCOME MEASURE(S): In the study group, mean ovarian volume, number of follicles, and number of oocytes recovered from each ovary were assessed and compared. The overall results, cycle characteristics, and pregnancy rates of the two groups were compared. RESULT(S): Among the patients who had undergone salpingectomy, significantly fewer follicles developed and consequently fewer oocytes were retrieved from the ovary on the operated side (4.4 versus 8.2 follicles and 3.8 versus 6.0 oocytes). There were no differences in the total numbers of follicles and oocytes recovered from both ovaries, the cycle characteristics, or the pregnancy rates between study and control groups. CONCLUSION(S): Salpingectomy has no detrimental effect on the total ovarian performance during IVF-ET treatment or on the outcome of IVF-ET. However, the ipsilateral ovary could be adversely affected. This could be detrimental in selected patients undergoing IVF ET, in whom the second ovary already is compromised or missing. PMID- 9848292 TI - Different influence of incongruent follicular development on in vitro fertilization-embryo transfer and gamete intrafallopian transfer pregnancy rates. AB - OBJECTIVE: To compare the influence of incongruent (asymmetric) follicular development on treatment outcome in IVF-ET and GIFT cycles. DESIGN: A retrospective comparative study. SETTING: Tertiary referral center for infertility. PATIENT(S): Five hundred forty-three consecutive assisted reproduction cycles (428 IVF-ET and 115 GIFT) in 422 infertile patients. INTERVENTION(S): Controlled ovarian hyperstimulation (COH) and IVF-ET or GIFT. MAIN OUTCOME MEASURE(S): The incongruity ratio as a parameter of the asymmetry in follicular development and pregnancy rate (PR). RESULT(S): For GIFT cycles, the PRs were 37.8% and 15.7% in cycles with congruent and incongruent follicular development, respectively. However, for IVF-ET cycles, the PR was not affected by incongruent follicular development: 28.2% and 29.0%, respectively. An inverse relationship was observed between the degree of incongruity and the estimated probability of pregnancy in GIFT cycles but not in IVF-ET cycles. Neither the side of the dominant ovary nor the degree of incongruity were consistent in consecutive cycles. CONCLUSION(S): Incongruent follicular development during COH has a significantly negative influence on the outcome of GIFT cycles but not on the outcome of IVF-ET cycles. The reason for this difference is not clear. We recommend considering IVF-ET instead of GIFT if incongruent follicular development occurs. PMID- 9848293 TI - Prednisolone plus low-dose aspirin improves the implantation rate in women with autoimmune conditions who are undergoing in vitro fertilization. AB - OBJECTIVE: To evaluate the effect of prednisolone plus low-dose aspirin (PSL/LDA) in women with autoimmune conditions who were enrolled in an IVF-ET program. DESIGN: A retrospective clinical study. SETTING: In vitro fertilization unit, Niigata University Hospital, Niigata, Japan. PATIENT(S): Three hundred seven women who underwent IVF-ET between January 1996 and December 1997. INTERVENTION(S): Prednisolone (10 mg/d) and aspirin (81 mg/d) were administered to the women with autoantibodies who chose to participate. MAIN OUTCOME MEASURE(S): Pregnancy and implantation rates with IVF-ET. RESULT(S): Women undergoing IVF who had positive antinuclear antibodies, with or without antiphospholipid antibodies, had significantly lower pregnancy and implantation rates than did women without autoantibodies (14.8% versus 21.7% and 6.8% versus 10.4%, respectively). The administration of PSL/LDA to women with antinuclear antibodies significantly improved the outcome of IVF-ET (40.6% pregnancy rate and 20.3% implantation rate). CONCLUSION(S): A high proportion of women who are undergoing IVF-ET have autoantibodies, which are associated with poor IVF outcomes. The administration of PSL/LDA to these women may improve their implantation rate. PMID- 9848294 TI - Ovarian stimulation and borderline ovarian tumors: a case-control study. AB - OBJECTIVE: To assess the risk of borderline ovarian cancer among infertile women treated with fertility drugs. DESIGN: Case-control study. SETTING: Nationwide data obtained from public registers and postal questionnaires. PATIENT(S): All Danish women <60 years old with borderline ovarian cancer during the period 1989 1994 and randomly selected population controls. The analysis included 231 cases and 1,721 controls. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Influence of parity, infertility, and fertility drugs on the risk of borderline ovarian cancer after multivariate confounder control. RESULT(S): The odds ratio (OR) for borderline ovarian cancer among infertile untreated nulliparous women compared with fertile nulliparous women was 1.9. The OR for borderline ovarian cancer among treated nulliparous women compared with untreated infertile nulliparous women was 1.5, and the OR among treated parous women compared with untreated infertile parous women was 1.5. CONCLUSION(S): Among fertile women, the difference in the risk of borderline ovarian cancer between nulliparous women and parous women was not statistically significant. Nulliparous women who were infertile and who did not receive medical treatment had a twofold higher risk of borderline ovarian cancer than fertile nulliparous women. There was no statistically significant increase in the risk of borderline ovarian cancer among nulliparous women who were treated with fertility drugs compared with nulliparous untreated infertile women or among parous women who were treated with fertility drugs compared with parous untreated infertile women. PMID- 9848295 TI - Selective early elimination of luteal support in assisted reproduction cycles using a gonadotropin-releasing hormone agonist during ovarian stimulation. AB - OBJECTIVE: To determine if women undergoing GnRH agonist-hMG stimulated IVF cycles can undergo successful discontinuation of luteal phase support. DESIGN: A protocol for selective discontinuation of luteal phase support was evaluated prospectively in women undergoing assisted reproduction cycles. SETTING: A tertiary care institutional-based assisted reproduction program. PATIENT(S): One hundred eighty-eight women who conceived after an IVF or zygote intrafallopian transfer cycle including a GnRH agonist between January 1994 and June 1997. INTERVENTION(S): Women with serum progesterone levels of > or = 60 ng/mL at 4 weeks' gestation were selected for discontinuation of their luteal phase support. MAIN OUTCOME MEASURE(S): Delivery rate. RESULT(S): Sixty-three women (62.4%) met the criteria for discontinuation of luteal phase support. There were no differences in the mean age, peak E2 levels, number of follicles, number of embryos transferred, or delivery rates (85.7% versus 78.9%) between the women who did and those who did not have discontinuation of their progesterone supplementation. CONCLUSION(S): These data reveal that luteal phase support can be discontinued successfully for selective women undergoing IVF who are receiving a GnRH agonist. PMID- 9848297 TI - Obstetric outcome of in vitro fertilized pregnancies complicated by severe ovarian hyperstimulation syndrome: a multicenter study. AB - OBJECTIVE: To assess the outcome of pregnancies conceived with the use of IVF that are complicated by severe ovarian hyperstimulation syndrome (OHSS). DESIGN: A retrospective nationwide multicenter study. SETTING: Sixteen of 19 tertiary care medical centers in Israel. PATIENT(S): All patients undergoing IVF who were hospitalized for severe OHSS between January 1987 and December 1996. MAIN OUTCOME MEASURE(S): Pregnancy rate (PR) and rates of multiple gestation, miscarriage, ectopic pregnancy, obstetric complications, and intervention. RESULT(S): A total of 163 patients who had severe OHSS after IVF treatment were identified, of whom 142 (87.1%) had undergone ET. The clinical PR was 73.2%; 42.3% were singletons, 33.6% were twins, 17.3% were triplets, and 6.7% were quadruplets. The miscarriage rate was 29.8%, whereas the incidence of ectopic pregnancy was 1.9%. Forty-four percent of all births were premature, and 62.1% of all newborns had low birth weight. The most common antenatal complications were pregnancy-induced hypertension (13.2%), gestational diabetes (5.9%), and placental abruption (4.4%). The rate of cesarean section was 44.1%. CONCLUSION(S): Among patients who have severe OHSS after IVF treatment, the pregnancy rate and the rates of multiple gestation, miscarriage, prematurity, low birth weight, pregnancy-induced hypertension, gestational diabetes, and placental abruption are significantly higher than those reported previously for pregnancies conceived with the use of assisted reproductive techniques. PMID- 9848298 TI - Insulin-like growth factor-1 and insulin-like growth factor-binding protein-3 in women with premenstrual syndrome. AB - OBJECTIVE: To determine whether women with premenstrual syndrome (PMS) have aberrations of the GH axis as has been demonstrated in individuals with depression. DESIGN: Prospective trial. SETTING: Department of Obstetrics and Gynecology, University of California, Los Angeles. PATIENT(S): After prospective screening, 32 healthy women with PMS and 32 asymptomatic controls completed the study. INTERVENTION(S): Subjects completed a daily PMS symptom diary and a Beck Depression Inventory. They underwent phlebotomy 5 days and 12 days after the LH midcycle surge, which was identified with the use of a urinary LH detection kit. MAIN OUTCOME MEASURE(S): Serum levels of insulin-like growth factor-1 (IGF-1), insulin-like growth factor-binding protein-3 (IGFBP-3), E2, and progesterone. RESULT(S): Levels of IGF-1, IGFBP-3, E2, and progesterone did not differ between women with prospectively documented PMS and control subjects. CONCLUSION(S): Premenstrual syndrome and affective disorder share common symptoms and possibly a common cause. Biochemical markers such as alterations in the somatotropic system often are associated with major depression. Levels of IGF-1 and IGFBP-3 did not differ between women with PMS and control subjects, supporting the concept that PMS and endogenous depression are biologically distinct entities. PMID- 9848296 TI - Effects of pretreatment with an oral contraceptive on the time required to achieve pituitary suppression with gonadotropin-releasing hormone analogues and on subsequent implantation and pregnancy rates. AB - OBJECTIVE: To assess the effect of pretreatment with an oral contraceptive (OC) on ovarian cyst formation during pituitary suppression with buserelin acetate. DESIGN: Prospective randomized trial. SETTING: Academic medical center. PATIENT(S): Eighty-three patients who were undergoing IVF-ET treatment. INTERVENTION(S): Patients in the study group were pretreated with an OC for 14 days starting on the first day of menstruation. The administration of SC buserelin acetate was initiated on the last day of OC administration. Patients in the control group began to receive buserelin acetate on day 2 of menstruation. Hormonal assays and ultrasound scans were performed on the first day of menstruation, and 7, 11, and 14 days after the commencement of buserelin acetate administration. Thereafter, these tests were performed weekly until pituitary suppression was achieved. MAIN OUTCOME MEASURE(S): Incidence of cyst formation. RESULT(S): A cyst developed in 27 patients in the control group (52.9%) and no patients in the study group (odds ratio [OR]=115; 95% confidence interval [CI]=10 617). Patients in the study group achieved pituitary suppression faster (median difference [MD]=7 days; 95% CI=4-14) and required fewer ampules of gonadotropin (MD=10; 95% CI=6-14). They recruited more follicles (MD=3; 95% CI=0-5) and had higher pregnancy rates (37.2% versus 33.3%). CONCLUSION(S): Pretreatment with an OC abolishes ovarian cyst formation, shortens the time required to achieve pituitary suppression, and decreases gonadotropin requirements without having a negative effect on pregnancy rates. PMID- 9848299 TI - Activated partial thromboplastin time is a predictive parameter for further miscarriages in cases of recurrent fetal loss. AB - OBJECTIVE: To determine whether clinically routine clotting tests such as activated partial thromboplastin time (aPTT), prothrombin time (PT), or fibrinogen can be used to predict further miscarriages. DESIGN: Prospective study. SETTING: Nagoya City University Hospital, Nagoya, Japan. PATIENT(S): A total of 261 patients with a history of two consecutive first-trimester spontaneous abortions who had no antiphospholipid antibodies or other autoimmune diseases and no anatomic anomalies were examined for aPTT, PT, and fibrinogen before becoming pregnant again. INTERVENTION(S): Blood tests were performed before pregnancy. Patients then were followed up during subsequent pregnancy and their outcomes were compared with their previous blood test results. MAIN OUTCOME MEASURE(S): Activated partial thromboplastin time, PT, and fibrinogen were measured by coagulation time methods. RESULT(S): Fifty-eight of 261 patients (22.2%) had a subsequent miscarriage. Mean (+/-SD) values for preconception aPTT in individuals whose subsequent pregnancies ended in success and failure were 88.2%+/-23.4% and 99.3%+/-26.4%, respectively. The difference was statistically significant. Respective values were 106.8%+/-22.8% and 106.3%+/-21.4% for PT and 245+/-61.1 mg/dL and 259.1+/-57 mg/dL for fibrinogen. These findings were not significantly different. CONCLUSION(S): A shortened aPTT before conception is associated with further miscarriages in patients with a history of recurrent spontaneous abortions who have no antiphospholipid antibodies. PMID- 9848300 TI - Randomized, double-blind, placebo-controlled study of the effects of raloxifene and conjugated equine estrogen on plasma homocysteine levels in healthy postmenopausal women. AB - OBJECTIVE: To investigate the long-term effects of raloxifene on fasting plasma homocysteine levels in postmenopausal women compared with conjugated equine estrogen (CEE). DESIGN: Randomized, double-blind, placebo-controlled study. SETTING: Outpatient department of a university hospital. PATIENT(S): Fifty-two hysterectomized, healthy postmenopausal women. INTERVENTION(S): Oral raloxifene in two dosages (60 mg/d [n=13] and 150 mg/d [n=13]), oral CEE (0.625 mg/d [n=13], and placebo (n=13) were given for 24 months. MAIN OUTCOME MEASURE(S): Fasting plasma homocysteine concentrations. RESULT(S): Plasma homocysteine levels were not altered in the placebo group. After 12 months, a significant reduction versus baseline in the mean plasma homocysteine level (-16%) was found only in the raloxifene 150-mg group. The mean change in plasma homocysteine levels within this group also was significantly different from the changes versus baseline found in the placebo group (+2%) and the raloxifene 60-mg group (-2%), but not different from those found in the CEE group (-8%). After 24 months, plasma homocysteine levels were decreased significantly in the raloxifene 150-mg and CEE groups compared with both baseline (-13% and -10%, respectively) and placebo values (-15% and -11%, respectively). No significant change in plasma homocysteine levels was observed in the raloxifene 60-mg group. CONCLUSION(S): Raloxifene has a favorable, dose-related effect on plasma homocysteine levels in postmenopausal women. PMID- 9848302 TI - The performance of CA-125 measurement in the detection of endometriosis: a meta analysis. AB - OBJECTIVE: To assess the diagnostic performance of serum CA-125 measurement in the detection of endometriosis. DESIGN: Meta-analysis. SETTING AND PATIENT(S): Twenty-three studies comparing serum CA-125 levels and laparoscopically confirmed endometriosis. INTERVENTION(S): Serum CA-125 measurement and laparoscopy. MAIN OUTCOME MEASURE(S): Sensitivity and specificity of serum CA-125 measurement in the diagnosis of endometriosis with laparoscopy as the reference standard. RESULT(S): The estimated summary receiver operating characteristic curves showed that the performance of serum CA-125 measurement in the diagnosis of endometriosis grade I/IV is limited, whereas its performance in the diagnosis of endometriosis grade III/IV is better. CONCLUSION(S): Despite its limited diagnostic performance, we believe that the routine use of serum CA-125 measurement in patients with infertility might be justified. In contrast to laparoscopy, serum CA-125 measurement is an inexpensive test that is not a burden for the patient. It could identify a subgroup of patients who are more likely to benefit from early laparoscopy. Studies reporting on the mutual dependence between serum CA-125 measurement and data from the history and physical examination are needed. PMID- 9848301 TI - Abnormalities in the serum insulin-like growth factor-1 axis in women with hyperandrogenism. AB - OBJECTIVE: To study the insulin-like growth factor-1 (IGF-1) axis in hirsute women. DESIGN: Controlled clinical study. SETTING: Tertiary care institutional hospital. PATIENT(S): Forty hirsute women and 17 women with normal menstrual cycles. INTERVENTION(S): Basal and ACTH-stimulated samples were obtained, and sampling was repeated 1 (gonadal stimulation) and 21 (gonadal suppression) days after a single 3.75-mg IM dose of triptorelin. Controls did not receive triptorelin for ethical reasons. MAIN OUTCOME MEASURE(S): Serum GH, IGF-1, IGF binding protein-3 (IGFBP-3), insulin, glucose, total testosterone, sex hormone binding globulin, E2, and gonadotropin levels. Basal and ACTH-stimulated steroid precursors were measured. RESULT(S): Patients with idiopathic hirsutism were identified by normal serum androgen levels (n=17). Those with functional ovarian hyperandrogenism (n=15) were identified by an increase in the serum testosterone level that normalized during gonadal suppression, whereas those with functional adrenal hyperandrogenism (n=8) were identified by an initial increase in the testosterone level that persisted during gonadal suppression. The adrenal hyperandrogenism group had increased IGF-1 levels compared with the control, idiopathic hirsutism, and ovarian hyperandrogenism groups. Patients with ovarian hyperandrogenism had normal TGF-1 concentrations, but their IGFBP-3 concentrations were lower than those of controls. No differences were observed in GH levels between any of the groups. These results persisted when the influence of age was corrected for. CONCLUSION(S): The IGF-1 axis appears to be involved in the pathogenesis of hyperandrogenism, especially in patients with adrenal hyperandrogenism, who have a clear increase in IGF-1 levels. Moreover, patients with ovarian hirsutism have decreased IGFBP-3 concentrations, which might enhance IGF-1 bioavailability. PMID- 9848303 TI - Human preembryo development on autologous endometrial coculture versus conventional medium. AB - OBJECTIVE: To evaluate the effect of autologous endometrial coculture versus conventional medium on preembryo development. DESIGN: Controlled systematic clinical study. SETTING: University-based IVF center. PATIENT(S): Women with a history of failed IVF-ET with poor preembryo quality. INTERVENTION(S): Patients underwent a luteal phase endometrial biopsy. The tissue then was digested enzymatically, and the stromal and glandular cells were separated by differential sedimentation rates. These cells were cultured to confluence, released, and then cryopreserved until the patient's IVF-ET cycle. All normally fertilized oocytes then were allocated systematically to growth on autologous endometrial coculture or conventional medium until transfer on day 3. MAIN OUTCOME MEASURE(S): Preembryo blastomere numbers and cytoplasmic fragmentation rates were measured. RESULT(S): Forty-two women underwent 44 cycles of IVF-ET. In the morning on day 3, the mean (+/-SD) number of blastomeres and cytoplasmic fragments per preembryo on coculture compared with conventional medium was 5.9+/-1.5 versus 5.5+/-1.4 and 21%+/-13% versus 24%+/-11. At transfer the mean (+/-SD) number of blastomeres per preembryo on coculture was 7.4+/-1.3 versus 6.7+/-1.9 on conventional medium. CONCLUSION(S): There was a significant improvement in the mean (+/-SD) number of blastomeres per preembryo and decrease in the fragmentation rate for preembryos on autologous endometrial coculture compared with noncocultured preembryos from the same patient. PMID- 9848304 TI - Timed analysis of the nuclear maturation of oocytes in early preantral mouse follicle culture supplemented with recombinant gonadotropin. AB - OBJECTIVE: To analyze the effects of combined FSH and variable doses of LH on the nuclear maturity and capacity to resume meiosis in oocytes from preantral follicles from prepubertal mice. DESIGN: Prospective, randomized, and controlled in vitro laboratory experiment. SETTING: Academic research environment. INTERVENTION(S): Meiosis was studied after somatic cell removal or after stimulation with hCG plus epidermal growth factor in three culture conditions: maturation medium with FSH alone and with two different doses of LH. MAIN OUTCOME MEASURE(S): The nuclear maturation of the oocytes and the E2, progesterone, and alpha-specific inhibin content of the conditioned medium. RESULT(S): Somatic cell removal and hormonal stimulation were equally effective in inducing germinal vesicle breakdown, but the hormonal stimulus was essential for the completion of meiosis, which was maximal (70%) on day 13 of culture. Continuous addition of LH to FSH during the oocytes' growth made them more prone to spontaneous resumption of meiosis I but resulted in a higher proportion of oocytes reaching the completion of meiosis. Estradiol and progesterone measurements demonstrated that the presence of LH influences luteinization. CONCLUSION(S): In contrast to oocytes grown in vivo, cumulus cell removal by itself is an insufficient stimulus for oocytes cultured in vitro to complete meiosis. Timed stimulation with hCG and epidermal growth factor increases nuclear maturation rates. A maximum number of metaphase II oocytes are obtained after a 13-day in vitro growth period when LH is added to the maturation medium. PMID- 9848305 TI - Activin A stimulates meiotic maturation of human oocytes and modulates granulosa cell steroidogenesis in vitro. AB - OBJECTIVE: To test the hypothesis that activin A promotes in vitro human oocyte meiotic maturation while inhibiting steroid secretion by nonluteinized antral granulosa cells. DESIGN: Prospective randomized controlled study. SETTING: A university medical center. PATIENT(S): Nine women ranging in age from 31-44 years who were undergoing oophorectomy for nonovarian pathology. INTERVENTION(S): Analysis of meiotic maturation of oocytes and steroid secretion by granulosa cells cultured in the presence or absence of activin A. MAIN OUTCOME MEASURE(S): Germinal vesicle breakdown (GVBD) and attainment of metaphase II (MII) in oocytes, and progesterone and E2 secretion by granulosa cells. RESULT(S): Activin A significantly enhanced GVBD (91% vs. 65%) for control and maturation to MII (56% vs. 35% for control) of immature oocytes. Activin A significantly suppressed basal, and inhibin A-and FSH-stimulated progesterone and E2 secretion by nonluteinized granulosa cells. CONCLUSION(S): Activin A is a promoter of oocyte maturation in vitro and a modulator of granulosa cell steroidogenesis in culture. PMID- 9848306 TI - Reduction of adhesion formation by intraperitoneal administration of Arg-Gly-Asp containing peptides. AB - OBJECTIVE: To evaluate the ability of a variety of peptides containing the Arg Gly-Asp (RGD) sequence to reduce the formation of intraperitoneal adhesions in a rabbit model. DESIGN: Prospective, randomized, double-blinded study. SETTING: University-based laboratory. ANIMAL(S): New Zealand white rabbits. INTERVENTION(S): Administration of RGD-containing peptides. MAIN OUTCOME MEASURE(S): Reduction of adhesion information. RESULT(S): In initial studies, two RGD-containing peptides (3 or a 10 amino acid peptides) were administered via Alzet miniosmotic pump to the site of injury. Administration of either of these peptides significantly reduced adhesion formation, but the larger peptide was more efficacious and reduced variability in the response. Further studies then were conducted with RGD-containing peptides five to six amino acids in length. Administration of these peptides also significantly reduced adhesion formation in a standard rabbit model. Administration of three of these peptides in a viscous vehicle at the end of surgery was also effective in reducing adhesion formation. CONCLUSION(S): The most effective combination tested was RGD-containing peptide Gly-dser-Arg-Gly-Asp-Ser-Pro in a viscous, cremophor-containing vehicle. These studies demonstrate that administration of an RGD-containing peptide was effective in reducing adhesion formation in this model. PMID- 9848307 TI - The expression of soluble intercellular adhesion molecule-1 in endometriosis. AB - OBJECTIVE: To investigate the association between concentrations of soluble intercellular adhesion molecule-1 (ICAM-1) and interferon-gamma (IFN-gamma) with regard to the severity of endometriosis. DESIGN: Cross-sectional study. SETTING: Infertility unit in a university hospital. PATIENT(S): Seventy-one infertile patients who underwent laparoscopic evaluation were divided into two groups: 36 patients with endometriosis served as cases, and 35 patients without endometriosis served as control subjects. INTERVENTION(S): Peripheral blood and peritoneal fluid (PF) were collected from these patients during laparoscopic examination. MAIN OUTCOME MEASURE(S): Quantitative determination of levels of human ICAM-1 and IFN-gamma was performed with the use of an ELISA. The data were evaluated by Student's t-test, analysis of variance, and regression analysis. RESULT(S): Significantly elevated levels of soluble ICAM-1 were found in the sera of patients with endometriosis, especially those with advanced stages of the disease. Decreased IFN-gamma levels, which were negatively correlated with serum levels of soluble ICAM-1, also were noted in the PF of patients with endometriosis. CONCLUSION(S): The increased serum levels of ICAM-1 found in patients with endometriosis may indicate the presence of an active disease process. Further, the increased levels of soluble ICAM-1 in peripheral blood were inversely correlated with the concentrations of IFN-gamma in PF. The increased levels of soluble ICAM-1 in plasma may be associated with an immunologic feedback response that blocks further infiltration of immune cells. These findings may be of value in the diagnosis and evaluation of endometriosis. PMID- 9848308 TI - Human sperm endothelial nitric oxide synthase expression: correlation with sperm motility. AB - OBJECTIVE: To characterize the pattern of endothelial nitric oxide synthase (eNOS) expression on human spermatozoa and to determine whether sperm eNOS expression correlates with sperm function. DESIGN: Prospective, observational study. SETTING: University infertility clinic. PATIENT(S): Twelve nonazoospermic infertile men. INTERVENTION(S): Semen samples (n=12) obtained from nonazoospermic infertile men were fractionated on discontinuous Percoll gradients. Endothelial nitric oxide synthase staining on spermatozoa was correlated with sperm motility in Percoll gradient-fractionated spermatozoa. Endothelial nitric oxide synthase protein was detected with the use of a previously characterized monoclonal antibody. Control slides were incubated with preabsorbed antibody or mouse immunoglobulin G. MAIN OUTCOME MEASURE(S): Localization of eNOS on human spermatozoa and correlation between the pattern of sperm eNOS expression and sperm motility. RESULT(S): Morphologically normal spermatozoa exhibited postacrosomal and equatorial eNOS immunostaining. However, abnormally shaped spermatozoa often exhibited aberrant staining (in the midpiece and/or head region). A significant negative correlation was observed between the percentage of sperm with aberrant eNOS immunostaining and the percentage of motile sperm (r= .46). CONCLUSION(S): The specific localization of eNOS to human spermatozoa suggests that nitric oxide may be involved in normal sperm physiology. However, aberrant patterns of sperm eNOS expression are associated with decreased sperm motility, possibly through the generation of excessive cytotoxic oxidants. PMID- 9848309 TI - Plasma membrane integrity of cryopreserved human sperm: an investigation of the results of the hypoosmotic swelling test, the water test, and eosin-Y staining. AB - OBJECTIVE: [1] To examine the relationship between sperm membrane integrity and motion parameters before and after cryopreservation; [2] to determine the capacity of the membrane integrity tests to predict the outcome of cryopreservation in fertile and infertile men; and [3] to examine the degree of agreement between tail and head membrane integrity of testicular and ejaculated immotile sperm cryopreserved for intracytoplasmic sperm injection. DESIGN: Prospective study. SETTING: Academic tertiary care institution. PATIENT(S): Fertile donors and normozoospermic oligozoospermic, and asthenozoospermic subfertile men. INTERVENTION(S): Semen samples were cryopreserved and thawed for analysis. MAIN OUTCOME MEASURE(S): Sperm membrane integrity and computer-assisted motion parameters. RESULT(S): The hypoosmotic swelling test and water test had a significant and positive correlation in the fresh and cryopreserved ejaculates of all groups. The results of the hypoosmotic swelling test correlated positively with the percent motility in the fresh ejaculates of fertile and subfertile men. None of the membrane integrity tests correlated with the cryosurvival rate in any group. In the ejaculated and testicular samples with no postcryopreservation motility, the simultaneous assessment of hypoosmotic swelling test and eosin showed that of 33% sperm exhibiting coiling with the hypoosmotic swelling test, only 9% were eosin negative, whereas 24% were eosin positive. CONCLUSION(S): [1] The water test may be a simpler replacement for the hypoosmotic swelling test; [2] none of the membrane integrity tests predicted sperm motility after cryopreservation; and [3] there was a high degree of disagreement between the hypoosmotic swelling test and eosin in the samples with no postcryopreservation motility. PMID- 9848310 TI - The heparin-glutathione test: an alternative to the hypo-osmotic swelling test to select viable sperm for intracytoplasmic sperm injection. AB - OBJECTIVE: To evaluate the heparin-glutathione test (HEGLUT) for the selection of viable sperm for intracytoplasmic sperm injection (ICSI). DESIGN: A prospective study. SETTING: Department of Pediatrics, Obstetrics and Gynecology, University of Valencia and Instituto Valenciano de Infertilidad. PATIENT(S): Semen samples from healthy donors and patients with infertility. INTERVENTION(S): Sperm samples were kept in culture for different periods in Ham's F-10 medium supplemented or not supplemented with heparin, reduced glutathione (GSH), or a heparin-GSH mixture. Control and heparin-GSH-treated spermatozoa were injected into hamster oocytes. The HEGLUT and ICSI were performed. MAIN OUTCOME MEASURE(S): Sperm nuclear decondensation, progressive and nonprogressive motility, and male pronucleus formation. RESULT(S): The maximum proportion of sperm nuclear decondensation (28.7%+/-2.1% versus 2.6%+/-0.5% in the control group) was reached after 60 minutes of incubation in the presence of a heparin-GSH mixture. Differences in the percentages of progressive and nonprogressive motility among treatments and times of incubation, although statistically significant, were biologically negligible. No statistically significant differences were observed in the rate of sperm head decondensation (8.2% [4/49] versus 11.1% [6/54]) and male pronucleus formation (18.4% [9/49] versus 22.2% [12/541) after the injection of control and treated spermatozoa into hamster oocytes. CONCLUSION(S): The HEGLUT may offer an alternative to the hypo-osmotic swelling test for the selection of viable sperm for ICSI. PMID- 9848311 TI - Effect of storage temperature on sperm cryopreservation. AB - OBJECTIVE: To evaluate the influence of cryopreservation temperature on human sperm motility and morphology. DESIGN: Controlled study, investigator was blinded to the type of cryopreservation. SETTING: University-based andrology laboratory. PATIENT(S): Sixteen semen samples with normal motility and sperm count from men after a fertility work up. INTERVENTION(S): Semen aliquots were either stored in a mechanical freezer at -70 degrees C or in liquid nitrogen at -196 degrees C for 7 days or 3 months. Test yolk buffer was used as a cryoprotectant. With use of a programmable freezing unit, all samples were cooled at a controlled rate. MAIN OUTCOME MEASURE(S): Sperm motility and morphology. RESULT(S): After 7 days of cryopreservation, there was a greater decrease in sperm motility among specimens maintained at -70 degrees C than among those maintained at -196 degrees C (47% versus 39% decrease). The difference in sperm motility was even greater after 3 months of cryopreservation (72% versus 39% decrease). No difference in postthaw sperm morphology was detected among sperm preserved at -70 degrees C versus -196 degrees C. CONCLUSION(S): Sperm cryopreservation at -196 degrees C is superior to cryopreservation at -70 degrees C. Sperm can be stored at -70 degrees C for a short period of time with a relatively modest loss of motility. PMID- 9848312 TI - Noncommunicating accessory uterine cavity. AB - OBJECTIVE: To report a case of a noncommunicating accessory uterine cavity. DESIGN: Case report. SETTING: University-affiliated reproductive endocrinology practice. PATIENT(S): A 15-year-old nulligravida with increasing dysmenorrhea. INTERVENTION(S): Pelvic ultrasound, intravenous pyelogram, hysterosalpingogram, laparoscopy, laparotomy, and resection of noncommunicating accessory uterine cavity. MAIN OUTCOME MEASURE(S): Results of imaging studies, surgical examination, and resection of anomaly. RESULT(S): Complete resection of accessory cavity and resolution of dysmenorrhea. CONCLUSION(S): The patient had a mullerian anomaly in which the uterus contained two uterine cavities. One normal uterine cavity with communication to both fallopian tubes was present along with a noncommunicating, accessory uterine cavity. PMID- 9848313 TI - Triplet pregnancy with acardius acranius after preimplantation diagnosis. AB - OBJECTIVE: To report the first case of fetal malformation after preimplantation diagnosis for Duchenne muscular dystrophy (DMD). DESIGN: Case report. SETTING: Perinatal center in a university hospital. PATIENT(S): A conductor for DMD in her third pregnancy. INTERVENTION(S): Preimplantation diagnosis was performed in an outside hospital. In our center, a dichorionic triplet pregnancy with acardius acranius was diagnosed. The anastomosis between the "pump"-twin and the fetus with acardius was embolized with histoacryl to prevent worsening cardiac insufficiency of the "pump"-twin. MAIN OUTCOME MEASURE(S): Pregnancy outcome. RESULT(S): The anastomosis between the "pump"-twin and the fetus with acardius was occluded successfully. Premature preterm rupture of membranes led to rapid labor and delivery at 24 + 5 weeks' gestation. The smaller girl died of severe hyaline membrane disease, whereas the other infant had no major clinical problems and has developed well. CONCLUSION(S): There might be an association between embryo biopsy and fetal malformations. The setting up of a birth register after embryo biopsy is strongly recommended. PMID- 9848314 TI - Repeated transmission of X-linked ocular albinism type 1 by a carrier oocyte donor. AB - OBJECTIVE: To report the transmission of unsuspected X-linked ocular albinism in an oocyte donor program. DESIGN: Case report. SETTING: University medical center. PATIENT(S): A 24-year-old white female oocyte donor and the outcomes of three recipient pregnancies. INTERVENTION(S): Clinical assessment and molecular diagnostic tests. MAIN OUTCOME MEASURE(S): Mutation detection. RESULT(S): Demonstration of carrier status and multiple transmissions of a mutant allele. CONCLUSION(S): We describe the transmission of a mutant allele for X-linked ocular albinism from an unsuspected carrier female oocyte donor to three independent pregnancies. We emphasize the need for diligent inquisition to clarify any unusual history of ocular or constitutional signs that might signify an X-linked disorder. PMID- 9848315 TI - Comparison of sperm separation methods: effect on recovery, motility, motion parameters, and hyperactivation. AB - OBJECTIVE: To compare the Enhance (Percoll; Conception Technologies, San Diego, CA) and PureSperm (Gen X International, Madison, CT) sperm preparation methods with respect to recovery (percentage of motile sperm), motility (%), path and progressive velocities (microm/s), and hyperactivation (%). DESIGN: Comparison of sperm processing methods. SETTING: University medical center-based clinical andrology laboratory and infertility program. PATIENT(S): Twenty-five men who presented for semen analysis. INTERVENTION(S): Each of 25 semen specimens were divided and each aliquot was prepared using two different density gradient centrifugation methods. MAIN OUTCOME MEASURE(S): The motile sperm recovery, percent motility, motion parameters, and percent hyperactivation were measured for each semen specimen (n=25) before and after separation with the use of the two methods. RESULT(S): There was no difference in the percent motility and motile count between specimens prepared with Enhance (Percoll) and PureSperm and fresh specimens. Statistically significant differences were found (fresh versus test) in the velocities and in hyperactivation (PureSperm only), and no differences were found between the processing methods. CONCLUSION(S): PureSperm appears to be as effective as Percoll (Enhance) for the recovery of good, progressively motile sperm for use in IUI or other assisted reproductive techniques. PMID- 9848316 TI - Randomized clinical trial of two laparoscopic treatments of endometriomas: cystectomy versus drainage and coagulation. AB - OBJECTIVE: To assess the efficacy of two laparoscopic methods for the management of endometriomas with regard to pain relief, pregnancy rate, and disease recurrence. DESIGN: Prospective, randomized clinical trial. SETTING: Tertiary care hospital. PATIENT(S): Sixty-four patients with advanced stages of endometriosis. INTERVENTION(S): Patients were randomly allocated at the time of laparoscopy to undergo either cystectomy of the endometrioma (group 1) or drainage of the endometrioma and bipolar coagulation of the inner lining (group 2). MAIN OUTCOME MEASURE(S): Pain relief and pregnancy rate. RESULT(S): Thirty two patients were enrolled in each group. The 24-month cumulative recurrence rates of dysmenorrhea, deep dyspareunia, and nonmenstrual pelvic pain were lower in group 1 than in group 2 (dysmenorrhea: 15.8% versus 52.9%; deep dyspareunia: 20% versus 75%; nonmenstrual pelvic pain: 10% versus 52.9%). The median interval between the operation and the recurrence of moderate to severe pelvic pain was longer in group 1 than in group 2 (19 months [range, 13.5-24 months] versus 9.5 months [range, 3-20 months]). The 24-month cumulative pregnancy rate was higher in group 1 than in group 2 (66.7% versus 23.5%). CONCLUSION(S): For the treatment of ovarian endometriomas, a better outcome with a similar rate of complications is achieved with laparoscopic cystectomy than with drainage and coagulation. PMID- 9848317 TI - Firm oysters year round? Activation of developmental programs in human oocytes. PMID- 9848318 TI - Firm oysters year round? Activation of developmental programs in human oocytes. PMID- 9848319 TI - Medical versus surgical treatment of endometriosis? PMID- 9848320 TI - ICSI for selected oocytes within a batch--save for the 2nd time around? Intracytoplasmic sperm injection. PMID- 9848321 TI - Relationship of anti-FSH antibodies to ovarian failure. PMID- 9848322 TI - Effect of granulosa cell apoptosis on fecundity? PMID- 9848323 TI - Effect of ovarian steroid production on adrenal steroidogenesis? PMID- 9848324 TI - Effect of ovarian steroid production on adrenal steroidogenesis? PMID- 9848325 TI - Towards a unified specialty: unity holds the key to diversity. PMID- 9848326 TI - Cancer in the elderly. PMID- 9848327 TI - Cancer in developing countries: Part II--Cancer control: strategies and priorities. PMID- 9848328 TI - Conformal radiotherapy: a clinical review. PMID- 9848329 TI - Paclitaxel and cisplatin as salvage treatment in patients with non-seminomatous germ cell tumour who failed to achieve a complete remission on induction chemotherapy. AB - The purpose of this study was to evaluate the efficacy and toxicity of paclitaxel and cisplatin combination chemotherapy as salvage treatment in patients with non seminomatous germ cell tumour. Sixteen patients with histologically proven germ cell tumour, measurable disease and/or elevated serum tumour markers were eligible for the protocol. All patients had previously not achieved a complete remission (CR) to platinum-based induction chemotherapy and cytoreductive surgery. The treatment consisted of paclitaxel 175-225 mg/m2 as a 3-hour infusion, followed by cisplatin 100 mg/m2, repeated every 3 weeks for up to four cycles. Seven patients achieved a marker-positive partial remission (PR) by the end of the cisplatin-based induction chemotherapy; the remainder had disease progression at the start of the paclitaxel plus cisplatin treatment. One (6%) CR and 3 (19%) PRs were achieved, with an overall response rate of 25% (90% confidence interval 7-43). The duration of the CR is currently 9+ months; two PRs lasted 2 months. One patient with a PR has been lost to follow-up. During a median follow-up of 8 months (range 1-11), 12 patients died from the disease progression. The median survival for the whole group was 7 months. Toxicity was moderate, with neutropenia grade 3 occurring in 29% of patients, thrombocytopenia grade 1-3 in 29%, creatinine > 130 mmol/l in 36%, peripheral neuropathy grade 1-2 in 50%, and nausea and vomiting in 43%. Paclitaxel plus cisplatin showed modest activity, with an overall response rate of 31% in patients with poor prognosis who had not achieved a CR on induction chemotherapy. PMID- 9848330 TI - PVB chemotherapy in patients with recurrent or advanced dysgerminoma: a Phase II study of the EORTC Gynaecological Cancer Cooperative Group. AB - Dysgerminoma accounts for 1% of all ovarian cancers and for 50% of all ovarian germ cell malignancies. Low stage patients (50%) can be cured with local treatment. The aim of this trial was to study the objective tumour response rate and toxicity of PVB (cisplatin, vinblastine, bleomycin) chemotherapy in patients with pure advanced or recurrent dysgerminoma. Eighteen eligible patients with advanced dysgerminoma were entered into this study. Three patients had local bulky recurrence only; all the others also had metastatic disease. The median age at entry was 27 years (range 1348). Seventeen patients had had prior surgery and one had undergone prior radiotherapy. The WHO performance status was 0 in 12 patients, 1 in three patients, and 2 in three patients. The treatment consisted of: intravenous or intramuscular bleomycin 30 mg on days 2, 9 and 16, intravenous vinblastine 0.15 mg/kg on days 1 and 2, and intravenous cisplatin 20 mg/m2 on days 1-5. This regimen was given at 3-week intervals for a total of four cycles. Twelve patients obtained a complete response (66%), five a partial response (28%), and one could not be evaluated because radiotherapy was administered immediately after chemotherapy. After a median follow-up of 76 months (range 4 132), 14 (78%) patients were alive and well. Two died of disease progression, one of neutropenic septicaemia and one of lung fibrosis. No unusual toxicity was reported. Alopecia, as well as nausea and vomiting, were common. Leucopenia (78%), thrombocytopenia (17%) and infection (11%) were the other severe (grade 3 4) side effects. The PVB chemotherapy regimen is highly effective in patients with advanced ovarian dysgerminoma. However, the BEP (bleomycin, etoposide, cisplatin) regimen, which is equally as potent and less toxic, is preferred. PMID- 9848331 TI - A non-randomized comparison of two radiotherapy protocols in inoperable squamous cell carcinoma of the oesophagus. AB - We report an audit of two different telebrachytherapy schedules in inoperable carcinoma of the oesophagus. Between October 1990 and December 1996, 108 patients with a Karnofsky performance status > or = 50 were selected from our database on the basis of intention to treat by telebrachytherapy. Teletherapy in the low dose group L (55 patients) consisted of 35 Gy in 15 fractions over 3 weeks, while that in the high dose group H (53 patients) consisted of 50 Gy in 25 fractions over 5 weeks. The choice of teletherapy dose was based on physician preference. The high dose rate intraluminal radiotherapy that followed 2 weeks later was identical in both groups and consisted of two applications of 6 Gy, a week apart. The pretreatment disease characteristics of the patients in both arms were similar. Relief of dysphagia was obtained in 49% of the patients in group L and in 75% of those in group H (chi2: P = 0.004). The median dysphagia-free interval was 0 and 7 months in groups L and H respectively (log-rank: P = 0.06), while the median overall survival was identical at 8 months (log-rank: P = 0.21) for both groups. The probability of survival at 1, 2 and 5 years was 34.8% versus 35.8%, 14.5% versus 13.9% and 0% versus 10% for groups L and H respectively. Morbidity in the form of ulcers, strictures and fistulae were observed in 9%, 7% and 5% of patients in group L compared with 8%, 8% and 13% in groups H respectively. This audit suggests that the protocol used in group H, when compared with group L, results in a greater proportion of patients being rendered dysphagia free, with a statistical trend towards a greater sustainment of dysphagia relief on follow-up. PMID- 9848332 TI - An audit of antiemetic use with CMF chemotherapy. AB - A two-stage retrospective audit of the efficacy of antiemetics used with intravenous cyclophosphamide 600 mg/m2, methotrexate 40 mg/m2 and 5-fluorouracil 600 mg/m2 (CMF) chemotherapy for breast cancer has been performed in a single centre. The first audit stage examined emesis rates for 21 patients using a policy of metoclopramide and dexamethasone initially and reserving 5-HT3 antagonists for failure of this combination. Because of high failure rates, the policy was changed so that granisetron and dexamethasone were used in all patients. The second audit stage examined the effectiveness of this new policy in 28 patients. Nine of 21 patients (43%) in the first audit group changed from metoclopramide and dexamethasone because of nausea or vomiting. Emesis occurred in 15 out of 66 (23%) cycles of CMF when metoclopramide and dexmethasone were used. Only eight of 43 patients had emesis episodes with granisetron and dexamethasone (19%). Emesis occurred with only 16 of 206 CMF cycles with granisetron and dexamethasone (8%). The granisetron and dexamethasone combination is superior to metoclopramide and dexamethasone in the prophylaxis of CMF-induced nausea and vomiting, although it is more expensive. A reduced dose of granisetron, 1 mg rather than 3 mg, may allow similar control of emesis at reduced cost. PMID- 9848333 TI - Pelvic exenteration for advanced primary rectal cancer in male patients. AB - Between November 1987 and January 1996 pelvic exenteration for primary rectal cancer was carried out in 11 male patients. Two underwent a primary resection with subtotal cystectomy. In the remaining nine patients, the treatment commenced with a staging laparotomy and the fashioning of an end colostomy of the descending colon, followed by preoperative radiotherapy (50-56 Gy in 5 weeks). Total exenteration with uretero-ileo-cutaneostomy was carried out 4-6 weeks later. In two patients, the exenteration was performed despite the detection of hepatic metastases during the second laparotomy. The pathological staging was T4 in six and T3 in five patients. The immediate postoperative course was uneventful in eight patients and their hospital stay averaged 20 days. Serious postoperative complications prolonged the hospital stay of three patients. Three died during the first 6 months. Recurrent disease caused the death of three of the eight remaining patients: one locoregional (7.5 years after surgery), one from pre existing hepatic metastases (18 months after surgery) and one from the combination of locoregional and distant recurrence (15 months after surgery). Four patients are on follow-up without evidence of disease, 96, 43, 23 and 22 months after surgery. One patient is alive 20 months after exenteration, with two pulmonary metastases having been recently excised. We conclude that, notwithstanding the morbidity rate, total pelvic exenteration is an acceptable option for advanced primary rectal cancer in male patients. Preoperative radiotherapy should be administered for cancers seated in the lower two-thirds of the rectum and perioperative chemotherapy should be considered seriously. PMID- 9848334 TI - Sister Joseph's nodule in non-Hodgkin's lymphoma. AB - Sister Joseph's nodule is traditionally regarded as a sign of poor prognosis in untreatable malignancy. We describe a patient with a metastatic umbilical nodule in non-Hodgkin's lymphoma who had a good outcome. The possible mechanisms of tumour spread to the umbilicus and reasons for its rarity in lymphoma are discussed. The need for histological confirmation to exclude treatable disease is emphasized. PMID- 9848335 TI - Syringomyelia after chemotherapy and radiotherapy for advanced oropharyngeal carcinoma: cause or coincidence? AB - A patient presented with syringomyelia 18 months after the completion of treatment with both chemotherapy and radiotherapy for an advanced oropharyngeal carcinoma. The link with therapy is discussed. PMID- 9848336 TI - Plasmablastic lymphoma: a new subcategory of human immunodeficiency virus-related non-Hodgkin's lymphoma. AB - A patient with human immunodeficiency virus-related non-Hodgkin's lymphoma arising in the oral cavity is reported. The tumour had an unusual immunohistochemical profile that was negative for leucocyte common antigen and the B-cell antigen CD20 but positive for the plasma cell-reactive antibody VS38c. The features of this type of tumour, which has recently been categorized, are presented and discussed. PMID- 9848337 TI - Intracranial juvenile nasopharyngeal angiofibroma. AB - We report the case history of a 26-year-old man who was diagnosed with advanced juvenile nasopharyngeal angiofibroma with intracranial extension. The lesion was considered to be inoperable and the patient was treated with radical radiotherapy. Serial magnetic resonance imaging has shown continued tumour regression and he remains well after 3 years. The literature is reviewed and radiotherapy recommended as the modality of choice for these patients. PMID- 9848338 TI - Neurological complications of 5-fluorouracil chemotherapy: case report and review of the literature. AB - Neurological complications of 5-fluorouracil (5-FU) chemotherapy are uncommon events. The two patients presented demonstrate two manifestations of 5-FU neurotoxicity, namely a cerebellar syndrome in association with global motor weakness and bulbar palsy, and a bilateral third cranial (oculomotor) nerve palsy. Both highlight the rapid onset and severity of these unusual side effects but also emphasize that, unlike paraneoplastic syndromes or central nervous system involvement by tumour, complete spontaneous recovery is a potential outcome. PMID- 9848339 TI - Neuropathic back pain: discordance between the level of the involved spine/rib and the associated dermatome. PMID- 9848340 TI - Imaging investigation protocols for childhood cancer. PMID- 9848341 TI - Evolution and development of hospice and specialist palliative care services. PMID- 9848342 TI - In-flight incidents. PMID- 9848343 TI - New susceptibility gene for Alzheimer's disease on chromosome 12? PMID- 9848344 TI - Oral or parenteral therapy for B12 deficiency. PMID- 9848345 TI - Aortic complications of Marfan's syndrome. PMID- 9848346 TI - Patient-information leaflets and prescriber competence. PMID- 9848347 TI - Changing patterns of mortality across Europe in patients infected with HIV-1. EuroSIDA Study Group. AB - BACKGROUND: The introduction of combination antiretroviral therapy and protease inhibitors has led to reports of falling mortality rates among people infected with HIV-1. We examined the change in these mortality rates of HIV-1-infected patients across Europe during 1994-98, and assessed the extent to which changes can be explained by the use of new therapeutic regimens. METHODS: We analysed data from EuroSIDA, which is a prospective, observational, European, multicentre cohort of 4270 HIV-1-infected patients. We compared death rates in each 6 month period from September, 1994, to March, 1998. FINDINGS: By March, 1998, 1215 patients had died. The mortality rate from March to September, 1995, was 23.3 deaths per 100 person-years of follow-up (95% CI 20.6-26.0), and fell to 4.1 per 100 person-years of follow-up (2.3-5.9) between September, 1997, and March, 1998. From March to September, 1997, the death rate was 65.4 per 100 person-years of follow-up for those on no treatment, 7.5 per 100 person-years of follow-up for patients on dual therapy, and 3.4 per 100 person-years of follow-up for patients on triple-combination therapy. Compared with patients who were followed up from September, 1994, to March, 1995, patients seen between September, 1997, and March, 1998, had a relative hazard of death of 0.16 (0.08-0.32), which rose to 0.90 (0.50-1.64) after adjustment for treatment. INTERPRETATION: Death rates across Europe among patients infected with HIV-1 have been falling since September, 1995, and at the beginning of 1998 were less than a fifth of their previous level. A large proportion of the reduction in mortality could be explained by new treatments or combinations of treatments. PMID- 9848348 TI - Prognostic value of minimal residual disease in acute lymphoblastic leukaemia in childhood. AB - BACKGROUND: Sensitive techniques for detection of minimal residual disease (MRD) at degrees of one leukaemic cell per 10(3)-10(6) cells (10(-3)-10(-6)) during follow-up of children with acute lymphoblastic leukaemia (ALL) can provide insight into the effectiveness of cytotoxic treatment. However, it is not yet clear how information on MRD can be applied to treatment protocols. METHODS: We monitored 240 patients with childhood ALL who were treated according to national protocols of the International BFM Study Group. 60 patients relapsed and the patients in continuous complete remission (CCR) had a median event-free follow-up of 48 months. Bone-marrow samples were collected at up to nine time points during and after treatment. Standardised PCR analysis of patient-specific immunoglobulin and T-cell receptor gene rearrangements and TAL1 deletions were used as targets for semiquantitative estimation of MRD. Amount of MRD was classed as 10(-2) or more, 10(-3), and 10(-4) or less. FINDINGS: MRD negativity at the various follow up times was associated with low relapse rates (3-15% at 3 years), but five-fold to ten-fold higher relapse rates (39-86% at 3 years) were found in MRD-positive patients. The distinct degrees of MRD appeared to have independent prognostic value (p [trend]<0.001) at all separate time points, especially at the first two time points (at the end of induction treatment and before consolidation treatment). At these two time points a high degree of MRD (> or = 10(-2)) was associated with a three-fold higher relapse rate when compared with patients with a low degree of MRD (< or = 10(-4)). At later time points (including the end of treatment) even a low degree of MRD was associated with a poor outcome. Positivity in patients in CCR after treatment was rare (< 1%). With the combined MRD information from the first two follow-up time points, it was possible to recognise three different risk groups--55 (43%) were in a low-risk group and had a 3-year relapse rate of only 2% (95% CI 0.05-12%); 19 (15%) were in a high-risk group and had a relapse rate of 75% (55-95%); and 55 (43%) were in an intermediate-risk group and had a 3-year relapse rate of 23% (13-36%). INTERPRETATION: Our collaborative MRD study shows that monitoring patients with childhood ALL at consecutive time points gives clinically relevant insight into the effectiveness of treatment. Combined information on MRD from the first 3 months of treatment distinguishes patients with good prognoses from those with poor prognoses, and this helps in decisions whether and how to modify treatment. PMID- 9848349 TI - Acute parvovirus B19 infection associated with fulminant hepatitis of favourable prognosis in young children. AB - BACKGROUND: The cause of fulminant hepatitis (FH) in children is unexplained in up to 50% of cases. We report parvovirus B19 as an agent associated with FH in children and compare clinical characteristics of these patients with those of age matched patients with FH of other origin. METHODS: 45 patients presented with FH. No cause was apparent in 21 patients. Parvovirus B19 genome was retrospectively sought by PCR in serum collected at admission in 41 patients. FINDINGS: Parvovirus B19 genome was detected in serum from four of 21 patients with unexplained FH (four of 11 younger than 5 years). No B19 DNA was detected in serum from patients with other types of FH or from 82 patients with biliary atresia. Parvovirus B19 IgM was detected in one of the four patients. Patients with parvovirus B19 infection had significantly lower bilirubin concentrations than age-matched patients with FH due to hepatitis A (nine) or other causes (nine) (poisoning with amanita excluded). All patients with parvovirus B19 survived without orthotopic liver transplantation, with restoration of normal liver function within 17 days. INTERPRETATION: In patients younger than 5 years with FH of unexplained origin, evidence of acute parvovirus B19 was associated with a distinct clinical pattern. In particular, low bilirubin concentrations and rapid recovery of liver function without transplantation were distinctive features. PMID- 9848350 TI - Acquired increase in leucocyte binding by intestinal microvascular endothelium in inflammatory bowel disease. AB - BACKGROUND: Endothelial cells that line microvascular blood vessels have an important role in inflammation through their ability to bind and recruit circulating leucocytes. Endothelial cells from the intestines of patients with chronically inflamed Crohn's disease and ulcerative colitis--the two forms of inflammatory bowel disease--display an increased leucocyte-binding capacity in vitro. We investigated whether this enhanced leucocyte binding is a primary or an acquired defect. METHODS: We cultured human intestinal microvascular endothelial cells (HIMEC) from the uninvolved intestine and chronically inflamed bowel of three patients with inflammatory bowel disease (two Crohn's disease, one ulcerative colitis). We assessed HIMEC binding to polymorphonuclear leucocytes and U937 cells by means of an adhesion assay. FINDINGS: After activation with interleukin-1beta or lipopolysaccharide, HIMEC from the chronically inflamed tissue in all three patients with inflammatory bowel disease bound twice as many polymorphonuclear leucocytes and U937 cells as endothelial cells from uninvolved tissue. INTERPRETATION: Enhanced leucocyte binding by HIMEC from chronically inflamed tissue in patients with inflammatory bowel disease is an acquired defect since it is not found in the uninvolved intestinal segments from the same individuals. Because interaction between endothelial cells and leucocytes is a key regulatory step in the inflammatory process, this enhanced binding may contribute to the pathophysiology of chronic intestinal inflammation. PMID- 9848351 TI - Increased fetoplacental angiogenesis during first trimester in anaemic women. AB - BACKGROUND: Epidemiological studies describe an association between relative size of the placenta at delivery and cardiovascular morbidity and mortality during adult life. Some determinants of placental size, such as maternal anaemia, have been acknowledged, but no plausible mechanism has been advanced to explain the initiation of postnatal disease. METHODS: Placental villous vascularisation in anaemic women (Hb<90 g/L) was assessed in the first and third trimesters of pregnancy by immunohistochemical identification of villous capillaries and compared with that of gestational age-matched groups of women with normal (Hb>110 g/L; control group) concentrations of haemoglobin, and an intermediate group (Hb 90-110 g/L). FINDINGS: Anaemia, especially in the first trimester, was associated with increased numbers of capillaries per villous cross section (mean 11.70 [SE 0.35] vs 4.14 [0.27]) located mainly in the outer third of the stroma beneath the trophoblast (94% [1.15] vs 67% [1.82]) and with increased numbers of villous macrophages and of proliferating MIB-1-positive cells compared with the control group. INTERPRETATION: Maternal anaemia in early pregnancy seems to influence the pattern of placental vascularisation. Such changes might alter placental vascular impedance during early fetal life, thereby exerting important effects on cardiovascular development. PMID- 9848352 TI - A 6-year-old child with vacant episodes and unilateral convulsions. PMID- 9848353 TI - Totally implantable hearing device for sensorineural hearing loss. PMID- 9848354 TI - Fatal interaction between ritonavir and MDMA. PMID- 9848355 TI - Reporting of safety data from randomised trials. PMID- 9848356 TI - Severe bone pain after intravenous pamidronate in adult patients with cystic fibrosis. PMID- 9848357 TI - Increased temporal cortex CREB concentrations and antidepressant treatment in major depression. PMID- 9848358 TI - Minocycline in early diffuse scleroderma. PMID- 9848359 TI - Homozygous arginine-72 in wild type p53 and risk of cervical cancer. PMID- 9848360 TI - Thrombophilic disorders in children with cerebral infarction. PMID- 9848361 TI - Indinavir concentrations in hair from patients receiving highly active antiretroviral therapy. PMID- 9848362 TI - What is a normal red-blood cell mass for professional cyclists? PMID- 9848363 TI - Remedy presented for health inequalities in England. PMID- 9848364 TI - Bad news for the young in AIDS epidemic update. PMID- 9848365 TI - Phyto-oestrogens: the way to postmenopausal health? PMID- 9848366 TI - Plans for UK Food Safety Agency go into reverse. PMID- 9848367 TI - Medical confidentiality broken to stop marriage of man infected with HIV. PMID- 9848368 TI - Principles of epidemiological research on adverse and beneficial drug effects. PMID- 9848369 TI - Evolution of the management of acute myocardial infarction: a 20th century saga. PMID- 9848370 TI - Involvement of platelets in tumour angiogenesis? AB - Preclinical and clinical research show that tumour growth is dependent on angiogenesis. Activation of the coagulation cascade is commonly found in patients with cancer. We propose that platelets contribute to tumour-induced angiogenesis. The basis of our hypothesis is that platelets are a rich source of stimulators and inhibitors of angiogenesis and their interaction with the endothelium. Presumably, the antithrombotic state of normal endothelium is disturbed by endothelial stimuli derived from tumour cells. This hypothesis may explain the suggested clinical benefits of anticoagulants in cancer and implies that targeting of platelet interaction with tumour vasculature will inhibit angiogenesis. PMID- 9848371 TI - Better health in developing countries: are sector-wide approaches the way of the future? PMID- 9848372 TI - Junior doctors and stress. PMID- 9848373 TI - Junior doctors and stress. PMID- 9848374 TI - Junior doctors and stress. PMID- 9848375 TI - Doping in sport. PMID- 9848376 TI - Lipid lowering therapy in patients with HIV infection. PMID- 9848377 TI - Lipid lowering therapy in patients with HIV infection. PMID- 9848378 TI - Surveillance of antimicrobial resistance. PMID- 9848379 TI - Subsets within the chemotherapy overview. International Breast Cancer Study Group. PMID- 9848380 TI - Changes in MTHFR genotype frequencies over time. PMID- 9848381 TI - Malignant hyperthermia. PMID- 9848382 TI - Malignant hyperthermia. PMID- 9848383 TI - Ischaemic heart disease in primary immunodeficiency. PMID- 9848384 TI - Neurotic disorder in India. PMID- 9848385 TI - Do we always need to tell patients the truth? PMID- 9848386 TI - Do we always need to tell patients the truth? PMID- 9848387 TI - Do we always need to tell patients the truth? PMID- 9848389 TI - Sabbaticals for UK doctors. PMID- 9848388 TI - Ethical issues in genetics of mental disorders. PMID- 9848390 TI - Sour grapes? PMID- 9848391 TI - The Nobel chronicles. 1928: Charles Jules Henry Nicolle (1866-1936). PMID- 9848392 TI - Sketches from The Lancet. Radiography. PMID- 9848393 TI - Enhanced clearance of a multiple antibiotic resistant Staphylococcus aureus in rats treated with PGG-glucan is associated with increased leukocyte counts and increased neutrophil oxidative burst activity. AB - PGG-Glucan [Betafectin], a highly purified soluble beta-(1-6)-branched beta-(1 3) linked glucan isolated from Saccharomyces cerevisiae, has broad in vitro and in vivo anti-infective activities unrelated to cytokine induction. Here we present in vivo results on the anti-infective activity of PGG-Glucan against a multiple antibiotic resistant Staphylococcus aureus. PGG-Glucan (0.25-4 mg/kg) was administered intramuscularly to male Wistar rats 48 h, 24 h, and 4 h before and 4 h after intraperitoneal implantation of a gelatin capsule containing 10(8)S. aureus colony forming units (CFU). Blood samples were collected at various times after challenge to determine CFU levels, leukocyte counts and neutrophil oxidative burst activity; serum TNF-alpha, and IL-1beta levels were also evaluated. The 0.25 mg/kg PGG-Glucan dose had no effect on reducing blood CFU levels; however, PGG-Glucan doses of 0.5 mg/kg, 1 mg/kg, 2 mg/kg or 4 mg/kg significantly reduced blood CFU levels by 48 h after challenge. Reduced CFU levels correlated with significantly elevated absolute monocyte counts, absolute neutrophil counts, and neutrophil oxidative burst activity in the absence of any effect on TNF-alpha or on IL-1beta levels. In additional studies, effects on mortality and blood CFU levels were evaluated in rats treated with ampicillin (an antibiotic to which the S. aureus was resistant), PGG-Glucan, or both agents. Mortality and blood CFU levels were reduced most in combination-treated rats compared to saline control rats or rats treated with either ampicillin alone or PGG-Glucan alone. We conclude that in vivo (1) PGG-Glucan can enhance clearance of an antibiotic resistant S. aureus, (2) that this clearance is accompanied by an increase in monocytes and neutrophils as well as a potentiation of neutrophil oxidative microbiocidal activity without alteration of the proinflammatory cytokine response, and (3) PGG-Glucan can enhance the effectiveness of traditional antibiotic treatment. PMID- 9848394 TI - A comparative investigation of the restorative effects of roxithromycin on neutrophil activities. AB - The effects of roxithromycin, a macrolide antibiotic, on neutrophil activities were investigated in six seriously handicapped patients with severe mental retardation. Neutrophil activities were evaluated by flow cytometry using a heparinized blood analysis method. All six patients showed decreased levels of neutrophil phagocytosis, intracellular killing, and CD11b expression. Treatment with roxithromycin in vitro selectively restored the decreased phagocytic and bactericidal activities of neutrophils in these patients. There was no significant restorative effect with cefaclor, ofloxacin, or aztreonam. These results suggest the need to consider therapeutic effects of antibiotics on neutrophil functions in patients at increased risk for bacterial infections due to decreased neutrophil activities. PMID- 9848395 TI - Inhibitory effect of ginsenoside on the mediator release in the guinea pig lung mast cells activated by specific antigen-antibody reactions. AB - We reported that some components of ginsenosides decreased mediator release which was evoked by the activation of mast cells caused by specific antigen-antibody reactions. This study aimed to assess the effects of ginsenoside, Rb1, which belongs to the protopanaxadiol, on the mechanism of mediator release during mast cell activation. Pretreatment of Rb1 (100 microg) significantly decreased histamine and leukotriene in a dose-dependent manner during mast cell activation. The PLD activity during mast cell activation decreased in the pretreatment of Rb1 (300 microg). The amount of DAG produced by PLC activity decreased because of Rb1 pretreatment. The amount of mass DAG decreased due to Rb1 pretreatment during mast cell activation. Rb1 (300 microg) pretreatment strongly inhibited the incorporation of the [3H]methyl moiety into phospholipids. The data suggest that Rb1, purified from Korean Red Ginseng Radix, inhibits an increase of DAG production during mast cell activation caused by antigen-antibody reactions, which is mediated via phosphatidylcholine-PLD and phosphatidylinositol-PLC systems. This is then followed by the inhibition of histamine releases. Furthermore, Rb1 reduces the phosphatidylcholine production by inhibiting the methyl-transferase I and II, and the reduction of phosphatidylcholine production inhibits leukotriene release. PMID- 9848397 TI - Anti-chemotactic activities of peptide-T: a possible mechanism of actions for its therapeutic effects on psoriasis. AB - Peptide T is an octapeptide (ASTTTNYT) from the V2 region of gpl20 of HIV. D [Ala]-Ser-Thr-Thr-Thr-Asn-Tyr-Thr-amide (DAPTA) is one of its analogue. DAPTA has been shown to resolve the psoriatic lesions. The mechanisms of action of peptide T for its therapeutic effect is not clearly understood. Lymphomononuclear cells play an important roles in inflammatory disease processes. Intraepidermal collection of lymphocytes is a unique feature of the inflammatory processes of psoriasis. It is believed that chemokine such as RANTES (C-C class) plays an important role for intraepidermal localization of the inflammatory infiltrates in psoriasis. In order to study the mechanisms, we have analyzed the effects of DAPTA on monocyte and lymphocyte chemotaxis. Chemotaxis of cells was measured by using Boyden chamber. DAPTA inhibited significantly the monocyte and lymphocyte chemotactic activity of RANTES (p < 0.005, < 0.001). Antichemotactic activities of peptide T analogue could be a possible explanation for its therapeutic efficacy in psoriasis. PMID- 9848396 TI - Cinnamaldehyde inhibits lymphocyte proliferation and modulates T-cell differentiation. AB - Two kinds of cinnamaldehyde derivative, 2'-hydroxycinnamaldehyde (HCA) and 2' benzoxy-cinnamaldehyde (BCA), were studied for their immunomodulatory effects. These compounds were screened as anticancer drug candidates from stem bark of Cinnamomum cassia for their inhibitory effect on farnesyl protein transferase activity. Ras activation, which is accompanied with its farnesylation, has been known to be important in immune cell activation as well as in carcinogenesis. Treatment of these cinnamaldehydes to mouse splenocyte cultures induced suppression of lymphoproliferation following both Con A and LPS stimulation in a dose-dependent manner. A dose of I microM of HCA and BCA inhibited the Con A stimulated proliferation by 69% and 60%, and the LPS-induced proliferation by 29% and 21%, respectively. However, the proliferation induced by PMA plus ionomycin was affected by neither HCA nor BCA treatment. Decreased levels of antibody production by HCA or BCA treatment were observed in both SRBC-immunized mice and LPS-stimulated splenocyte cultures. The exposure of thymocytes to HCA or BCA for 48 h accelerated T-cell differentiation from CD4 and CD8 double positive cells to CD4 or CD8 single positive cells. The inhibitory effect of cinnamaldehyde on lymphoproliferation was specific to the early phase of cell activation, showing the strongest inhibition of Con A- or LPS-stimulated proliferation when added concomitantly with the mitogens. In addition, the treatment of HCA and BCA to splenocyte cultures attenuated the Con A-triggered progression of cell cycle at G1 phase with no inhibition of S to G2/M phase transition. Although cinnamaldehyde treatment had no effect on the IL-2 production by splenocyte cultures stimulated with Con A, it inhibited markedly and dose-dependently the expression of IL-2Ralpha and interferon-gamma. Taken together, the results in this study suggest both HCA and BCA inhibit the lymphoproliferation and induce a T-cell differentiation through the blockade of early steps in signaling pathway leading to cell growth. PMID- 9848398 TI - Paclitaxel reduces anti-dsDNA antibody titer and BUN, prolonging survival in murine lupus. AB - We evaluated the effect of paclitaxel on the severity of autoimmunity in the murine model of systemic lupus erythematosus (SLE), NZB x NZW F1 mice. Fifteen 20 week old (NZB x NZW) F1 female mice were given a dose of 10 mg/kg paclitaxel by the intraperitoneal route on three alternate days followed by 7.5 mg/kg on three additional alternate days. This pattern of treatment was repeated every 4 weeks for a period of 28 weeks. 20 control mice were injected intraperitoneally with an equal volume of the vehicle used. Serum anti-double stranded DNA (dsDNA) antibody titers and the blood urea nitrogen (BUN) were significantly diminished in the paclitaxel treated group compared to the vehicle treated group. While the onset of proteinuria appeared to be delayed in the experimental group, the difference was not significant. Survival rate improved significantly in paclitaxel treated group (p = 0.04 by log-rank test). These results suggest that paclitaxel is beneficial in the suppression of autoimmunity in this strain of mice by reducing the anti-dsDNA antibody titer and the BUN, prolonging survival. PMID- 9848399 TI - Federated healthcare record server--the Synapses paradigm. AB - The delivery of healthcare relies on the sharing of patient information between those who are providing for the care of the patient and this information is increasingly being expressed in terms of a 'record'. Further, it is desirable that these records are available in electronic form as Electronic HealthCare Records. As it is likely that patient records or parts of records will be stored in many different information systems and in the form of disparate record architectures, uniform access to patient records would be problematic. This paper presents an overview of the Synapses computing environment in which a Federated Healthcare Record Server provides uniform access to patient information stored in connected heterogeneous autonomous information systems and other Synapses servers. The Synapses record architecture is based on the architecture proposed by the Technical Committee 251 of the European Committee for Standardisation and the interfaces to the Synapses server are specified in the ISO standard Interface Definition Language. Synapses is a pan-European project involving a number of hospitals, software companies, universities and research institutes and is partly funded by the EU Health Telematics Programme. The overview is described in terms of the Open Distributed Processing Reference Model. PMID- 9848400 TI - CORBA security services for health information systems. AB - The structure of healthcare systems in developed countries is changing to 'shared care', enforced by economic constraints and caused by a change in the basic conditions of care. That development results in co-operative health information systems across the boundaries of organisational, technological, and policy domains. Increasingly, these distributed and, as far as their domains are concerned, heterogeneous systems are based on middleware approaches, such as CORBA. Regarding the sensitivity of personal and medical data, such open, distributed, and heterogeneous health information systems require a high level of data protection and data security, both with respect to patient information and with respect to users. This paper, relying on experience gained through our activities in CORBAmed, describes the possibilities the CORBA middleware provides to achieve application and communication security. On the background of the overall CORBA architecture, it outlines the different security services previewed in the adopted CORBA specifications which are discussed in the context of the security requirements of healthcare information systems. Security services required in the healthcare domain but not available at the moment are mentioned. A solution is proposed, which on the one hand allows to make use of the available CORBA security services and additional ones, on the other hand remains open to other middleware approaches, such as DHE or HL7. PMID- 9848401 TI - The standard 'Healthcare Information Systems Architecture' and the DHE middleware. AB - The integration and evolution of existing systems represents one of the most urgent priorities of health care information systems in order to allow the whole organisation to meet the increasing clinical organisational and managerial needs. This paper discusses how an open architecture, based on the introduction of a middleware of common health care-specific services not only reduces the effort necessary for allowing existing systems to interwork, but also automatically establishes a functional and information basis common to the whole organisation, on top of which also new applications can be rapidly developed, natively integrated with the rest of the system. Such architecture has been already formalised through the European standard, defined by the CEN/TC251 prENV 12967-1 'Architecture for Health care Information Systems' (CEN prENV 12967-1 'Health care Information Systems Architecture'). Thanks to the availability of industrial products conforming to the standard, the effectiveness and the validity of this approach has been already demonstrated in practice. For example, through the Hansa collaboration hospitals and industries from countries of the Western and Eastern Europe, as well as of the Middle East use the same industrial middleware (i.e. 'The DHE middleware-Information view'-SPRI, 1998, 'The DHE middleware Functional view'-SPRI, 1998) for integrating existing systems as well as for developing new applications. PMID- 9848402 TI - Component-based development for supporting workflows in hospitals. AB - Changing requirements for health care information systems force the development of an open, modular architecture in which components can be integrated. This offers a flexible means for integrating different (heterogeneous) systems used by different users. Selecting the components to integrate, and determining the 'right way' to integrate them, necessitates a shift in focus towards the business process to be supported. The realization of such an open, modular architecture is a difficult task. It consists of breaking down existing systems in required components and integrating these and other components. Many authors on component based development strategies focus attention on the technological issues of component integration (do we use CORBA, DCOM/OLE, or EDI?). This paper presents an approach for determining the required components, and the way they have to be integrated, based on an analysis of the business process to be supported, and the information systems currently used. PMID- 9848403 TI - Communication technologies in health care environments. AB - A successful implementation of hierarchical health care networks is dependent on a number of different technical and non technical factors. This paper investigates how the networking technologies, both 'traditional' and broadband, can support the communication requirements of medical environments. The general purpose is to implement communication infrastructures and services, which will improve the collaboration between the different partners in the health sector in offering better healthcare services at affordable costs. PMID- 9848404 TI - Computer based nursing documentation means to achieve the goal. AB - New functions have been integrated in the Giessen Hospital Information System WING to support the classification of all intensive care patients into the Therapeutic Intervention Scoring System (TISS). The use of those functions has been pushed when health insurance bodies demanded evidence for the correct classification of ICU beds. This article presents an overview on this development from the start in just one intensive care unit to the complete coverage of six intensive care units and three intensive monitoring units with a total of 109 beds. For those units complete TISS data has been documented for more than a year now at a detailed level. On average 14 interventions have been recorded per patient and day, accumulating to a database with more than a million entries. We describe the experiences made during introduction and the different front-end applications we used to achieve the goal. Results gained from the huge database and their implications for our future work are discussed. TISS documentation is now an established routine on every intensive care unit of our University hospital. It has been implemented without major financial or manpower investments and no specific intensive care information system has been needed. Establishing this type of basic care documentation made nurses aware of their activities, so that now they consider electronic care documentation to be in their very own interest. The next goal has been set by nurses themselves, they want to establish intervention based care documentation on normal wards as well. We think that step by step we will thus be able to achieve a more complete electronic patient record. PMID- 9848405 TI - Telematics techniques for image based diagnosis, therapy planning and monitoring. AB - This paper is intended to describe and illustrate some of the actual use of telematics related techniques together with modern biomedical imaging capabilities for helping in diagnosis, as well as for the planning and monitoring of therapy. To this end, most current imaging modalities are initially introduced. Then it is shown how telematics related techniques are necessary to improve the outcome of current image-based protocols. Such techniques allow data, means, or competencies--which may intrinsically be of a complementary nature or distributed at many different locations--to be integrated together and transcend the simple sum of individual expectations. Examples of actual implementations are given in the fields of radio-oncology, neurosurgery and orthopedics. To conclude, the papers and posters presented in the corresponding session of the MIE'97 symposium are summarized to provide further telematics references for the reader. PMID- 9848406 TI - Medical emergency aid through telematics: design, implementation guidelines and analysis of user requirements for the MERMAID project. AB - MERMAID is an EU financed telemedicine project with global reach and 24-h, multilingual capability. It aspires to provide a model for the provision of health care services based on the electronic transmission of medical information, via ISDN based videoconferencing. This model will not be limited to medical diagnostics but it will encompass all cases where the actual delivery of health care services involves a patient who is not located where the provider is. Its implementation requires the commissioning of an expensive telecommunications infrastructure and the exploration of a number of solutions. In fact, all categories of telemedical applications (audio and video conferencing, multimedia communications, flat file and image transfer with low, medium and high bandwidth data requirements) are considered while the full range of network choices (digital land lines, cellular/wireless, satellite and broadband) are being tested in terms of cost/performance tradeoffs that are inherent to them and the developmental stage each of these options occupies in their in its life cycle. Finally, out that MERMAID utilises advanced land based line transmission technologies to aid the remote patient by making available the specialist care that is best suited in the particular case. PMID- 9848407 TI - Towards personal health record: current situation, obstacles and trends in implementation of electronic healthcare record in Europe. AB - In this article, we define the electronic healthcare record and present its purpose as a tool for continuity of care. We briefly describe the current situation of usage and focus on the major challenges to wide implementation in Europe and beyond. Finally, we point out trends that show stronger involvement of the patients-citizens in the health care prevention and promotion processes, and discuss the impact on the future development of the electronic healthcare record into personal health records. PMID- 9848408 TI - Protocols and guides, tools. AB - The author describes in the first part of the review the definitions and relations of the occurring concepts. In the second part a short history of the topic is overviewed. The third part is dealing with the electronic patient record based development of guidelines in addition what are the relations among guidelines, protocols, tools, knowledge-based systems, diagnosis supporting systems, expert systems, outcome analysis and artificial intelligence. A short review of what was presented in the MIE '97 in this area is finally described and the Session Chairman's conclusion. PMID- 9848409 TI - The basic principles of the synapses federated healthcare record server. AB - Synapses is a project funded under the EU Health Telematics Framework IV Programme. Synapses sets out to solve problems of sharing data between autonomous information systems, by providing generic and open means to combine healthcare records or dossiers consistently, simply, comprehensibly and securely, whether the data passes within a single healthcare institution or between institutions. This paper presents the specification of the Synapses server, the kernel concept of Synapses. It describes the basis in the European prestandard for Electronic Healthcare Record Architecture, the interfaces to the Synapses client and server and different integration mechanisms for systems providing information to the server. The specification will be verified at a number of validation sites and the final result will be in the public domain. PMID- 9848410 TI - Present and future trends with NLP. AB - This paper reflects some desiderata on the role of Natural Language Processing (NLP) in the coming years as foreseen in the medical domain. During the Medical Informatics Europe (MIE) conference in 1997, the NLP track was composed by numerous papers on natural language, knowledge representation, nomenclatures and classifications. Indeed, the medical community is looking for solutions for the future which are going to emerge from more powerful desktop and for which the successful softwares of tomorrow are not yet identified. Presently this same community's needs remains unsatisfied with healthcare professionals writing at best their patient medical records in a text processing system. We stand today closer to the typewriter than to any modern solution which could radically transform the treatment of information related to patients. What kind of hurdles are there still in front of us before we reach this new territory? PMID- 9848411 TI - Trends and pitfalls with nomenclatures and classifications in medicine. AB - This paper reflects some of the main points in the history and the trends of classifications, nomenclatures and knowledge representation models in medicine. The relations with philosophical background are briefly related. The contributions in this field during the Medical Informatics Europe (MIE) conference in 1997 are presented. International collaborations in the field of lexico-semantic resources and inter-disciplinary partnership in natural language processing technologies are necessary steps to achieve definitive results that will lead to applications in the daily medical practice. PMID- 9848412 TI - From syntactic-semantic tagging to knowledge discovery in medical texts. AB - In the GALEN project, the syntactic-semantic tagger MultiTALE is upgraded to extract knowledge from natural language surgical procedure expressions. In this paper, we describe the methodology applied and show that out of a randomly selected sample of such expressions coming from the procedure axis of Snomed International, 81% could be analysed correctly. The problems encountered fall in three different categories: unusual grammatical configurations within the Snomed terms, insufficient domain knowledge and different categorisation of concepts and semantic links in the domain and linguistic models used. It is concluded that the Multi-TALE system can be used to attach meaning to words that not have been encountered previously, but that an interface ontology mediating between domain models and linguistic models is needed to arrive at a higher level of independence from both particular languages and from particular domains. PMID- 9848413 TI - A review of compression methods for medical images in PACS. AB - As an introduction to Section C2 (medical imaging) of track C (Images and PACS) of MIE '97, an appropriate and timely topic concerns the coding for the transmission of medical images in PACS. Speed limitations of existing networks along with the explosive growth of image modalities with extremely high volume outputs have combined to make the issue of medical image coding one of the key considerations in the design of future PACS systems. Both lossless and lossy compression schemes are reviewed and compared, and the compression demands are presented of the main digital medical image modalities. PMID- 9848414 TI - A fast and accurate method for registration of MR images of the head. AB - This paper proposes a new fully automated technique that can be used for the registration of medical images of the head. The method uses Chebyshev polynomials in order to approximate and then minimize a novel multiresolutional, signal intensity independent disparity function, which can generally be defined as the mean squared value of the mean weighted ratio of two images. This function is explicitly computed for n Chebyshev points in a geometric transformation parameter interval [-A, +A] transformation units and is approximated using the Chebyshev polynomials for all other points in the interval. For 3D T2-T1 weighted MR registration, 120 experiments with studies from ten patients were performed and showed that n = 4 Chebyshev points for A = 18 transformation units give mean rotational error 0.36 degrees and a mean translational error 0.36 mm. The different noise conditions did not affect the performance of the method. We conclude that the method is suitable for routine clinical applications and that it has significant potential for future development and improvement. PMID- 9848415 TI - A wavelet-based reduction of heart sound noise from lung sounds. AB - Heart sounds produce an incessant noise during lung sounds recordings. This noise severely contaminates the breath sounds signal and interferes in the analysis of lung sounds. In this paper, the use of a wavelet transform domain filtering technique as an adaptive de-noising tool, implemented in lung sounds analysis, is presented. The multiresolution representations of the signal, produced by wavelet transform, are used for signal structure extraction. In addition, the use of hard thresholding in the wavelet transform domain results in a separation of the nonstationary part of the input signal (heart sounds) from the stationary one (lung sounds). Thus, the location of the heart sound noise (1st and 2nd heart sound peaks) is automatically detected, without requiring any noise reference signal. Experimental results have shown that the implementation of this wavelet based filter in lung sound analysis results in an efficient reduction of the superimposed heart sound noise, producing an almost noise-free output signal. Due to its simplicity and its fast implementation the method can easily be used in clinical medicine. PMID- 9848416 TI - ECG pattern recognition and classification using non-linear transformations and neural networks: a review. AB - The most widely used signal in clinical practice is the ECG. ECG conveys information regarding the electrical function of the heart, by altering the shape of its constituent waves, namely the P, QRS, and T waves. Thus, the required tasks of ECG processing are the reliable recognition of these waves, and the accurate measurement of clinically important parameters measured from the temporal distribution of the ECG constituent waves. In this paper, we shall review some current trends on ECG pattern recognition. In particular, we shall review non-linear transformations of the ECG, the use of principal component analysis (linear and non-linear), ways to map the transformed data into n dimensional spaces, and the use of neural networks (NN) based techniques for ECG pattern recognition and classification. The problems we shall deal with are the QRS/PVC recognition and classification, the recognition of ischemic beats and episodes, and the detection of atrial fibrillation. Finally, a generalised approach to the classification problems in n-dimensional spaces will be presented using among others NN, radial basis function networks (RBFN) and non-linear principal component analysis (NLPCA) techniques. The performance measures of the sensitivity and specificity of these algorithms will also be presented using as training and testing data sets from the MIT-BIH and the European ST-T databases. PMID- 9848417 TI - Education and health informatics. AB - In this contribution, the role of health informatics in the medical curriculum is discussed. Firstly, trends in healthcare are presented that may have an impact on the use of IT in healthcare and consequently on education. Then, the traditional educational system is discussed and it is argued that the educational system should be changed. The problem-based approach is presented as an example of a new approach. The implications for education and training in health informatics are given. Given the lack of knowledge with respect to the basics of health informatics of both health professionals and students, attention has to be paid to learning materials. Both IT-EDUCTRA and NIGHTINGALE are presented as European projects that focus on education and training. PMID- 9848418 TI - Trends in assessment of IT-based solutions in healthcare and recommendations for the future. AB - State-of-the-art assessments of IT-based solutions in Healthcare is discussed in this paper. Special emphasis is placed on the human and organisation-centred perspective at the development of IT-based solutions. Based on this and basic conditions for system analysis and design reported in the literature, requirements for a methodology for user-driven, constructive assessment during the entire life-cycle of the IT-based system are synthesised. Finally, the outcome is discussed in the light of the presentations on technology assessment during the MIE 97 Congress and other related literature. PMID- 9848419 TI - How can usability measurement affect the re-engineering process of clinical software procedures? AB - As a consequence of the dramatic improvements achieved in information technology standards in terms of single hardware and software components, efforts in the evaluation processes have been focused on the assessment of critical human factors, such as work-flow organisation, man-machine interaction and, in general, quality of use, or usability. This trend is particularly valid when applied to medical informatics, since the human component is the basis of the information processing system in health care context. With the aim to establish an action research project on the evaluation and assessment of clinical software procedures which constitute an integrated hospital information system, the authors adopted this strategy and considered the measurement of perceived usability as one of the main goals of the project itself: the paper reports the results of this experience. PMID- 9848420 TI - Organizational issues in health informatics: a model approach. AB - In this paper, we present a model that describes the stages of the implementation of an information system in a health care organization. The model offers no explanation of the implementation process but rather describes in a cyclic order the domains that are relevant when implementing a system. The model offers thus an opportunity to identify gaps in our knowledge and understanding of implementation processes and provides also the conceptual basis for a higher education course of health informatics that is focusing on organizational change and the pivotal role of information and communication technology. PMID- 9848421 TI - Considerations for sociotechnical design: experiences with an electronic patient record in a clinical context. AB - This paper argues that we should understand the process of IT design as the development of sociotechnical configurations. Drawing upon our experiences with an electronic patient record (EPR) on an Intensive Care Unit (ICU), we depict medical work practices as natural systems. Several considerations for design are developed. First, the EPR should not be overly structured with rationalistic and prefixed notions of the organization and content of medical work. Implementing structure is crucial, however, this should be derived from detailed, empirical knowledge of the practice involved. Second, it is crucial to ensure that the usage of the system will yield immediate benefits for primary users: the systems should support work, not generate it. Third, designing IT should include being aware of the socio political nature of seemingly 'neutral' tools as EPRs. PMID- 9848422 TI - Is our appearance important to our patients? AB - OBJECTIVES: We aimed to explore patients' attitudes towards family physicians' and nurses' appearance. METHODS: One hundred and sixty-eight patients from three teaching Family Medicine clinics in Beer-Sheva, Israel, were interviewed in the clinics regarding the medical staff's dress code. They were also asked to choose one picture for either a male or female physician which, in their opinion, was the most suited for their own family physician, from a selection of pictures of the same male and female doctors dressed in different attires. RESULTS: One hundred and twenty-six patients (75%) replied that the attire of the physician had no influence on their decision in choosing their own family doctor. Fifty-two per cent of the patients preferred the doctor in a white coat and 71 % had the same preference for the nurse. Older age was associated with increased preference for a white coat. The dressing items which scored high for male doctor were a name tag, a formal suit or a shirt with a tie and sports shoes. For a female doctor a name tag, short haircut, trousers and sports shoes ranked highly. Long hair, earrings, and sandals scored low for a male physician, while mini-dress, shorts and tight clothes scored low for a female physician. CONCLUSION: About half of patients still prefer the doctor to be dressed in a white coat. Patients prefer a more formal dressing for male and female physicians in family medicine clinics. Most of the patients claimed that the attire of the physician had no influence on their choice of family physician. PMID- 9848423 TI - Outcome in general medical patients presenting with common symptoms: a prospective study with a 2-week and a 3-month follow-up. AB - BACKGROUND: Although physical complaints account for over half of all ambulatory visits, surprisingly little is known about their natural history and factors affecting prognosis. OBJECTIVES: Our aims were to determine the outcome in general medical patients presenting with physical complaints and to delineate which factors impact upon recovery rates. METHODS: In this cohort study, 500 adults presenting to a general medicine clinic with a chief complaint of a physical symptom were interviewed in order to determine symptom characteristics, the presence of depressive or anxiety disorders, and expectations regarding the visit. Outcomes were assessed immediately post-visit and at 2 weeks and 3 months. The primary outcome was symptomatic improvement, and secondary outcomes included functional status, unmet expectations, satisfaction with care and visit costs. RESULTS: The majority (70%) of patients improved by 2 weeks follow-up and, of those who had not, 60% got better within 3 months. Moreover, relapse in patients initially better at 2 weeks was uncommon (6%) in the ensuing several months. While symptoms of recent onset had the highest improvement rates, half of those patients whose symptom had been present a year or longer also improved within 2 weeks. Improvement rates varied somewhat by symptom type, but no specific symptom had an improvement rate of less than 50%. Patients who had not improved at 2 weeks were more likely to report continuing serious illness worry, unmet expectations, functional impairment and dissatisfaction, even 3 months after the initial visit. CONCLUSION: Most general medical patients with physical complaints improve within 2 weeks of their initial clinic visit. Further attention may best be focused on the minority of patients who fail to improve and experience continuing concerns and impairment. PMID- 9848424 TI - Gender differences in general practice consultations: methodological challenges in epidemiological research. AB - BACKGROUND: Women consult their GP more often than men do. The distribution of complaints and diagnoses are different for women and men patients. Although several findings on gender differentials on mortality and health care consumption are rather consistent across studies, detailed findings and subsequent conclusions diverge in several important fields. OBJECTIVES: Our aim was to explore methodological aspects of research on gender differences in general practice. METHODS: We reviewed empirical studies within this field, aiming to identify methodological and interpretative intricacies which deserve special attention in epidemiological research on GP consultations. RESULTS AND CONCLUSIONS: We found that descriptive and explanatory levels of research are frequently confused. Simple questions, answers and explanations are commonly raised for complex issues within a poorly defined theoretical explanatory framework. There is a need to assess relevant approaches for various purposes, and to develop more uniform conceptual terms. Findings from one level are often transferred to another, incompatible level. Epidemiological issues must be considered, especially matters related to denominator level and standardization/confounders-not in order to decide which level represents 'reality', but to clarify the consequences of different measures for different research questions. The contents of the core variables and the potentials for bias should be discussed in order to provide a sound basis for future explanatory studies. PMID- 9848425 TI - Use of complementary and alternative medicine among primary care patients. AB - OBJECTIVE: Complementary and alternative medicine use is increasing worldwide, and the expenses are high while its effectiveness is still in debate. The aim of this survey was to evaluate the utilization of complementary and alternative medicine in Israel. METHOD: Four-hundred and eighty patients in two primary care clinics have participated in the survey and answered an anonymous questionnaire. RESULTS: Ninety patients (18.7%) have consulted an alternative medicine therapist at least once in the past. Both younger (0-19) and older (65 and older) age groups were associated with a low rate of complementary and alternative medicine utilization. In the adult population, an academic education was associated with a higher utilization rate. The most frequently used methods were homeopathy (34.6%) and reflexology (18.7 %). Musculo-skeletal (20.6%) and respiratory (15.9%) complaints were the most frequent causes for complementary and alternative medicine consultation. The subjective outcome of complementary and alternative medicine treatment was considered beneficial in almost half of the cases and partially beneficial in another 34.6%. CONCLUSIONS: Utilization of complementary and alternative medicine is as widespread in Israel as in other Western countries. Utilization rates were found to be associated with age and education but not with gender or origin. PMID- 9848426 TI - Communication between physicians and with patients suffering from breast cancer. AB - BACKGROUND: The communication between GP and specialists is vital for the patient suffering from breast cancer. OBJECTIVES: The aim of this study was to investigate (i) the speed and type of communication between GPs, specialists and patients with breast cancer, and (ii) the problems that GPs encounter in the communication with specialists concerning these patients. METHOD: In April 1995, 246 Dutch GPs from the Zwolle region (600 000 inhabitants) were invited to complete a questionnaire, using the information from the medical record and focusing on the last patient consulted with a confirmed diagnosis of breast cancer. RESULTS: Valid replies were received from 150 (61 %) GPs. The median period between initial referral date and receipt of the definite diagnosis from the surgeon was 4 weeks. After the patient's first appointment with the surgeon, the GPs received reports for 24% of the patients within 3 days; for 31% within 3 7 days; and for 16% of the patients after more than 2 weeks. After the first consultation between patient and surgeon, 68 (45%) of the 150 GPs reported that the patient contacted them; at this stage only 30 (20%) of these GPs had received a report from the surgeon. Thirty-one (21%) GPs did not contact the patient after receival of the definite diagnosis. GPs stated that the communication on patients with breast cancer is too slow (49%), or not frequent enough (25%); 25% of GPs found that the distribution of tasks between them and the specialists are not well described. CONCLUSION: In the diagnostic stage of breast cancer the communication between GPs, specialists and patients varies widely, is too slow and is incomplete. An effect of this unsatisfactory communication is that the patient herself is the messenger of the bad news. PMID- 9848427 TI - Assessment of GP management of symptoms of dying patients in an Australian community hospice by chart audit. AB - BACKGROUND: With specialist palliative care services becoming widespread, and the place of the GP in palliative care being examined, audit of patient care delivered by GPs is required in order to ensure adequate standards of care. OBJECTIVE: We aimed to evaluate symptomatic care delivered to palliative care patients by GPs in an Australian community hospice with a developed quality assurance programme. METHODS: The study was set in a newly established community based, GP-run hospice in a provincial city in Queensland, Australia. A chart audit was carried out of the first 20 patients admitted to a community-based hospice, in order to establish (i) whether attempts were made by the treating doctor to find the direct cause of symptoms before initiating management; and (ii) whether management accorded with developed consensus-based guidelines. RESULTS: Twenty patients were treated by 14 GPs; 135 new symptoms were identified in the records of these patients. Of the 125 symptoms for which guidelines could be identified in the literature, in 87 (70%) an attempt by the treating GP to find a direct cause could be demonstrated. Of the 114 symptoms with treatments defined in the guidelines, 107 (90%) treatments conformed to the guidelines. Constipation, nausea/vomiting, anorexia and back pain were the conditions for which there were the fewest attempts at establishing a direct cause before treatment. In most cases these conditions were treated in accordance with the guidelines. CONCLUSION: Quality assurance mechanisms present in an in-patient palliative care setting appear to be associated with high-quality care by GPs. PMID- 9848428 TI - Factors affecting practice nurse involvement in follow-up care of patients following myocardial infarction. AB - BACKGROUND: Preventive care can reduce the morbidity and mortality of patients following myocardial infarction. Recent evidence has shown that such care is not being provided effectively. The involvement of practice nurses has been proposed as a means of improving the completeness of follow-up and the quality of preventive care. OBJECTIVES: We aimed to determine the extent to which follow-up care for people discharged from hospital after a myocardial infarction is currently being undertaken by practice nurses and to assess the factors influencing the provision of such care and the nurses' attitudes towards this extended role. METHOD: Postal questionnaires were sent to 183 practice nurses working in general practices in the Southampton and South-West Hampshire Health District; 121 nurses responded (66%), representing 58 out of 64 practices surveyed (91%). RESULTS: The majority of responding practice nurses (55%, 95% CI 47-64%) had hospital experience of caring for patients with ischaemic heart disease, and most (83%, 95% CI 76-89%) believed that they played a key role in follow-up care of patients following myocardial infarction. In the absence of external support from a cardiac liaison nurse, few nurses (26%, 95% CI 16-41%) provide such care at present and only 21% work in practices with a register of myocardial infarction patients. Factors predicting the provision of follow-up care are having adequate time (odds ratio 4.59, 95% CI 1.66-12.7), the support of a cardiac liaison nurse (odds ratio 3.07, 95% CI 1.28-7.34) and GP colleagues (odds ratio 3.38, 95% CI 1.38-8.23), training in consultation skills (odds ratio 7.25, 95% CI 2.08-25.3), fundholding (odds ratio 3.11, 95% CI 1.26-7.69) and the confidence and knowledge of the practice nurse (odds ratios and 95% CIs respectively: 2.84, 1.18-6.83 and 2.80, 1.13-6.89). CONCLUSION: Most practice nurses are enthusiastic and have some of the necessary experience to provide follow-up care for patients who have experienced a myocardial infarction. Yet few currently provide it. The most important organizational incentives for providing such care are further training and the support of GPs and the cardiac liaison nurse. PMID- 9848429 TI - The early diagnosis of dementia: triggers, early signs and luxating events. AB - BACKGROUND: In order to make it possible and feasible that GPs diagnose dementia at an earlier phase than usual, it is important to know what elements in the available information on the patient and the family may trigger the diagnostic hypothesis of dementia. METHOD: By qualitative research, the stories of family members of dementia patients, according to DSM-III-R criteria, were analysed. RESULTS AND CONCLUSIONS: This work produced the following hypotheses: disturbances in functioning at work and ADL-IADL disturbances, the seeking of the carer's support and emotional problems-in addition to the more classical memory problems-are early signs and may be essential triggers for the GP to consider the possibility of an emerging dementia. Change of medication, narcosis, loss of the carer and hospital admission are luxating moments for the sudden appearance of symptoms and disturbances caused by dementia. Further research should be carried out to develop instruments for the early detection of dementia that step across neuropsychological tests and are feasible for daily use in a primary care setting. PMID- 9848430 TI - Attitudes of GPs to the care of people with epilepsy. AB - BACKGROUND: Most individuals with current epilepsy are solely under the care of the primary care team for follow-up care. Government working party recommendations, expert epilepsy panels and patients have also stressed the central role of the GP in follow-up care. Problems in the provision of care in the community have, however, repeatedly been highlighted. The views of GPs about service provision for people with epilepsy may be an important barrier to providing care, but have not yet been studied in a systematic manner. OBJECTIVES: We aimed to ascertain the views of GPs on service provision for people with epilepsy in primary care and on specific initiatives to improve care. METHOD: A specially designed postal questionnaire was sent to all 262 GPs on the list of West Glamorgan FHSA. It ascertained what GPs felt their role should be in providing care to people with epilepsy, identified their views on the importance of particular problems in providing this care, as well as obtaining their opinions on possible future initiatives to improve epilepsy care in the community. RESULTS: The overall response rate was 70%. Although a majority of responding GPs (55%) agreed that the care of people with epilepsy should be based in general practice, 23% disagreed. A lack of confidence about knowledge of epilepsy (34% responders), unfamiliarity with new drugs (65% responders) and a lack of time (41% responders) were identified as important perceived barriers to providing epilepsy care. Nearly all responding GPs would welcome guidelines for epilepsy care (93% felt they would be very helpful) and an epilepsy liaison nurse in the community was the most popular option in terms of preferred overall strategy for improving care. CONCLUSIONS: Despite 40 years of official recommendations regarding the central role of the GP in the follow-up care of people with epilepsy, a number of GPs have difficulty in providing this care. Many feel that they lack knowledge or are too time pressured to improve the situation. Nearly all GPs say that they would find guidelines for epilepsy care very helpful and over half would find epilepsy liaison nurses helpful. There is scope for more innovative ideas for epilepsy care in the community. PMID- 9848431 TI - Assessment of vaccine coverage of schoolchildren in three primary health care areas in rural Crete, Greece. AB - OBJECTIVE: A school-based immunization survey was used to assess vaccination coverage of a population of children in the region of three Primary Health Care Centres in rural Crete. METHODS: The medical charts of 771 (92.6% of the total) students attending primary schools at the municipalities of Archanes and Hersonissos in the district of Heraklion, and Neapolis in the district of Lasithi, as well as the charts of 438 (68.9% of the total) students attending the secondary school at Neapolis, Lasithi, were obtained. The immunization records from these charts were then compiled and analysed. All information was based on the standardized procedures of the Spili Health Centre, Crete. The age-specific vaccination rate was calculated for children of both primary and secondary school age. RESULTS: The vaccination coverage against diphtheria and tetanus was high (82%, 1035 pupils had received the fifth dose) for both groups of students, while the coverage for pertussis was incomplete, especially the fifth dose, which had been received by only 17% (n = 131) of the children of primary school age and 4.3% (n = 19) of the children of secondary school age. In total, 638 of the children of primary school age (82.7%) and 257 (58.7%) of the children of secondary school age were immunized against measles. The corresponding numbers and the percentages of children of primary and secondary school age immunized against mumps and rubella were: 75.6% (n = 583) and 36.3% (n = 159) for mumps, and 74.7% (n = 576) and 32% (n = 140) for rubella. CONCLUSION: Immunization coverage of schoolchildren in the studied areas is inadequate in a number of instances. This appears to be most pronounced in children of secondary school age. The school-based immunization survey, which is an accurate and inexpensive tool for assessing vaccination coverage, is suggested as an appropriate means for primary health care workers in Greece to develop further and increase the immunization rate. PMID- 9848432 TI - Concerns and cautions about prescribing and deregulating emergency contraception: a qualitative study of GPs using telephone interviews. AB - OBJECTIVES: We aimed to describe GPs' responses to a clinical scenario of a request for a repeat prescription for hormonal emergency contraception (EC), their views about over-the-counter availability and beliefs about absolute contraindications. DESIGN: We conducted semi-structured tape-recorded telephone interviews with 76 GPs randomly selected from the medical registers of three health authorities which were chosen for high, medium and low prescribing rates for EC. RESULTS: There was a wide variation in the number of times that GPs would be happy to prescribe EC to the same woman in a year. The content of the consultations appeared patchy. While 59 (77.6%) of the GPs said that they would discuss future contraception with the woman, only 16 (21.1%) said they would talk about possible side effects and 28 (36.3%) would discuss the timing of the next menstrual period and the possibility of method failure. Fifty-two of the practices had a family-planning-trained practice nurse, yet only four (7.7%) had arrangements whereby the nurse could provide EC. Unqualified enthusiasm for deregulation was rare. Concerns included that women would lose out on the benefits of the consultation; worries about the safety of the method; that some women might 'abuse' it by using it frequently; and that certain characteristics of the pharmacy might make it an unsuitable setting for provision of EC. CONCLUSIONS: This qualitative telephone survey revealed concerns about repeated use of EC and caution about the prospects of deregulation. Respondents were worried that pharmacists might not be able to address all of the features of the consultation that may be valued, yet in this sample nor do most GPs. Family planning-trained practice nurses are an under-utilized resource and could act as a halfway house between provision by GPs and deregulation. PMID- 9848433 TI - Is prophylactic treatment after myocardial infarction evidence-based? AB - OBJECTIVE: We aimed to evaluate the use of evidence-based prophylactic treatment after myocardial infarction on hospital discharge and in primary care after 1 year of hospitalization. METHODS AND RESULTS: We conducted a 1-year prospective study of all the patients discharged from a tertiary hospital who had been treated for myocardial infarction from January 1 to December 31 1995. Three hundred and eighty surviving patients were consecutively discharged from the hospital. Seventy per cent of patients were treated with aspirin, 45% with beta blockers, 27% with calcium channel blockers, 26% with ACE inhibitors, 40% with nitrates and 8% with cholesterol-lowering drugs after discharge from the hospital. In primary care, prescription of lipid-lowering drugs increased to 17%, while prescription of beta-blockers decreased to 34%. ACE inhibitor prescriptions at discharge were clearly more common in patients with impaired ventricular function or heart failure (57%). CONCLUSION: According to the evidence, there is still potential for reducing the risk of a further ischaemic event or death in patients with MI, especially by increasing the use of beta-blockers and lipid lowering drugs. PMID- 9848434 TI - A review of research in the United Kingdom to evaluate the implementation of clinical guidelines in general practice. AB - BACKGROUND: An advisory group for the NHS research and development (R&D) programme recommended in 1993 that the impact of clinical guidelines at the interface between primary and secondary care should be a research priority area. In 1994, a systematic review of 91 published evaluations of implementing clinical guidelines identified only seven UK general practice studies. OBJECTIVE: In this inquiry we aimed to determine the number of randomized studies of clinical guideline implementation in primary care being conducted in the UK in 1996 and to review the research designs. METHODS: A national health research register was interrogated for all projects relating to clinical guidelines. The investigators were contacted to establish the nature of their project and to identify implementation studies that they knew to be going on elsewhere. Copies of protocols or briefing documents were obtained from the project teams for the identified studies. RESULTS: Thirteen randomized studies in general medical practice and one in general dental practice were identified. Guidelines were being introduced to aid diagnostic decision-making, prescribing practice or referral to hospital-based services. Eight strategies for promoting guideline adherence were being evaluated. Six studies proposed to conduct economic evaluations of the intervention packages. CONCLUSION: Twelve of the 14 studies were funded by the NHS R&D programme. Since there will be a considerable time delay before all 14 studies and a number of newer studies are fully reported, it seems imperative that information of the sort collected in this inquiry be made available, preferably in the National Research Register. PMID- 9848436 TI - Doctor-patient communication and patient satisfaction: a review. PMID- 9848435 TI - A PIL for every ill? Patient information leaflets (PILs): a review of past, present and future use. AB - This article reviews the usefulness and importance of written information, specifically leaflets, being given to patients. Evidence suggesting how both patient and doctor may benefit from the giving of written information is reviewed. Identification of good practice relating to the content and readability of leaflets is discussed. An argument is put forward that the giving of written information is an under-utilized resource in contributing to improving patient outcomes but that this may be changing with the increasing use of patient leaflet databases. The advantages and disadvantages of computer-generated patient leaflets are discussed and desirable further areas of research on computer generated leaflets are proposed. PMID- 9848437 TI - Selections from current literature: androgenetic alopecia: the science behind a new oral treatment. PMID- 9848438 TI - Quorum sensing: a novel target for anti-infective therapy. PMID- 9848439 TI - Spiramycin renaissance. PMID- 9848440 TI - Evaluation of disc diffusion and Etest for determining the susceptibility of Staphylococcus aureus to mupirocin. AB - The susceptibilities to mupirocin of 102 selected clinical isolates of Staphylococcus aureus and of control strain S. aureus NCTC 6571 were determined by disc diffusion (using discs containing 5, 15, 25, 30, 50 and 200 microg of mupirocin) and Etest and the results were compared with MICs determined using an agar incorporation method. On the basis of agar incorporation MICs, 42 isolates were sensitive to mupirocin (MIC < or = 4 mg/L), 39 showed low-level resistance (MICs = 8-128 mg/L) and 22 were highly resistant (MICs > or = 256 mg/L) and contained the mupA resistance gene. Using Stokes' criteria, none of the discs used gave major errors (sensitive isolates classified as highly resistant) or very major errors (highly resistant isolates classified as sensitive) in assigning a category of susceptibility, but minor errors (a difference of one category) were noted with all strengths. The best correlation with agar incorporation MIC was obtained with 25 microg mupirocin discs, which classified correctly 98 (95%) isolates, while worse correlations were noted with 5 microg and 200 microg discs which are the only types currently available commercially, for which there were 47 and 30 minor errors, respectively. The MICs found by Etest were the same as, or lower than, those determined by agar incorporation. Etests classified correctly all 42 mupirocin-sensitive isolates, 19 (49%) low level resistant isolates and 16 (73%) highly resistant isolates. Two isolates that contained the mupA gene and showed agar incorporation MICs of 256 mg/L and 512 mg/L were not classified as highly resistant by any of the diffusion methods used. Agar incorporation MIC determination, possibly supported by detection of the mupA gene, offers the most effective means of identifying high-level mupirocin resistance in S. aureus, although the Etest also proved to be reproducible. However, we conclude that 25 microg discs warrant further evaluation for possible use in clinical laboratories, as they appear to be more reliable than the discs currently available. PMID- 9848441 TI - In-vitro and in-vivo susceptibility of Aspergillus fumigatus to a novel conjugated styryl ketone. AB - We investigated the in-vitro and in-vivo susceptibility of Aspergillus fumigatus to the novel conjugated styryl ketone NC1175 and the results were compared with those obtained for amphotericin B and itraconazole. All 20 clinical isolates of A. fumigatus examined were susceptible to NC1175 (MIC = 5.54 +/- 2.48 mg/L; range 2.92-11.68 mg/L), and the minimum lethal concentration (MLC) was only twice the MIC, suggesting that NC1175 is fungicidal. The mean MIC values of amphotericin B (1.22 +/- 0.58 mg/L; range 0.5-4 mg/L) and itraconazole (0.37 +/- 0.11 mg/L; range 0.125-0.5 mg/L) were approximately nine- and 22-fold, respectively, lower than that of NC1175. Both amphotericin B-resistant (n = 18) and itraconazole resistant (n = 28) isolates of A. fumigatus were as susceptible to NC1175 as amphotericin B-, and itraconazole-susceptible isolates. Kill curve experiments revealed that NC1175 at 23.35 mg/L (approximately four times the MIC) killed > or = 99% of conidia within 24 h of exposure to the drug. The in-vivo susceptibility of A. fumigatus to NC1175 was investigated using a murine pulmonary aspergillosis model. Treatment of infected mice with amphotericin B or NC1175 did not result in significant improvement of the mean survival (amphotericin B, 7.05 +/- 0.07 days; NC1175, 6.65 +/- 1.25 days) of the animals compared with that of the placebo group (7.21 +/- 1.20 days). However, semiquantitative organ culture revealed that clearance of A. fumigatus occurred in 16.6%, 50% and 66.6% of the mice treated with placebo, NC1175 and amphotericin B, respectively (P value for the control and the treated groups <0.01). These results suggest that NC1175 has in-vivo and in-vitro activity against A. fumigatus and can be used as a prototypic molecule for further development as an antifungal agent. PMID- 9848442 TI - In-vitro activity of essential oils, in particular Melaleuca alternifolia (tea tree) oil and tea tree oil products, against Candida spp. AB - The in-vitro activity of a range of essential oils, including tea tree oil, against the yeast candida was examined. Of the 24 essential oils tested by the agar dilution method against Candida albicans ATCC 10231, three did not inhibit C. albicans at the highest concentration tested, which was 2.0% (v/v) oil. Sandalwood oil had the lowest MIC, inhibiting C. albicans at 0.06%. Melaleuca alternifolia (tea tree) oil was investigated for activity against 81 C. albicans isolates and 33 non-albicans Candida isolates. By the broth microdilution method, the minimum concentration of oil inhibiting 90% of isolates for both C. albicans and non-albicans Candida species was 0.25% (v/v). The minimum concentration of oil killing 90% of isolates was 0.25% for C. albicans and 0.5% for non-albicans Candida species. Fifty-seven Candida isolates were tested for sensitivity to tea tree oil by the agar dilution method; the minimum concentration of oil inhibiting 90% of isolates was 0.5%. Tests on three intra-vaginal tea tree oil products showed these products to have MICs and minimum fungicidal concentrations comparable to those of non-formulated tea tree oil, indicating that the tea tree oil contained in these products has retained its anticandidal activity. These data indicate that some essential oils are active against Candida spp., suggesting that they may be useful in the topical treatment of superficial candida infections. PMID- 9848443 TI - Accumulation of rifampicin by Escherichia coli and Staphylococcus aureus. AB - A centrifugation method was used to investigate the accumulation of 14C rifampicin by Staphylococcus aureus and Escherichia coli, and to characterize the mechanism of rifampicin transport into S. aureus. For both species, drug accumulation was rapid with the steady-state concentration (SSC) reached within 40 s of drug exposure. Rifampicin accumulation by S. aureus was temperature and pH dependent; the lower the experimental temperature and the lower the experimental pH, the lower was the concentration of rifampicin accumulated. Accumulation was unaffected by the presence of inhibitors of antibiotic efflux, carbonyl cyanide m-chlorophenylhydrazone (CCCP), dinitrophenol (DNP), or reserpine. Exposure to increasing concentrations of rifampicin suggested that the accumulation process was saturable above a rifampicin concentration of 0.2 mg/L. Michaelis-Menten kinetics gave an apparent Km and Vmax for rifampicin, calculated from a Lineweaver-Burk plot, of 0.05 mg/L (0.06 microM) and 3.8 ng rifampicin per second, respectively. However, calculations suggest that these values reflect those for binding of rifampicin to its target, RNA polymerase. PMID- 9848444 TI - Molecular analysis of diverse elements mediating VanA glycopeptide resistance in enterococci. AB - Differences were examined among 24 distinct elements mediating VanA-type glycopeptide resistance in enterococci isolated from hospital patients and non human sources in the UK. The methods used included long-PCR restriction fragment length polymorphism (L-PCR RFLP) analysis and DNA hybridization. All elements had conserved vanRSHAX genes, but variation occurred upstream of vanR and downstream of vanX. Twenty-one VanA elements had significant alterations upstream of vanR in the transposition genes orf1 and orf2: either parts of these genes were absent or they were disrupted by IS1216V or IS3-like insertion sequences. Among VanA elements with alterations downstream of vanX, seven lacked vanY, one lacked both vanY and vanZ, and ten had copies of insertion sequence IS1216V between vanX and vanY. All VanA elements of group D (from geographically and temporally diverse enterococci) were characterized by the presence of an IS1216V/IS3-like/orf1 complex and a point mutation in vanX, both of which were absent from the other 23 groups of VanA elements. This finding is consistent with the dissemination of a stable resistance element. We conclude that L-PCR RFLP analysis, combined with DNA hybridization, merits further development for studying the evolution and epidemiology of VanA resistance elements in enterococci. PMID- 9848445 TI - The presence of the tetO gene in a variety of tetracycline-resistant Streptococcus pneumoniae serotypes from Washington State. AB - Seventy tetracycline-resistant Streptococcus pneumoniae were tested for the presence of tetracycline resistance genes, tetM and tetO, using a polymerase chain reaction (PCR) assay and DNA-DNA hybridization. Seven isolates representing five serotypes (12, 22, 6A, 19F and 23) carried the tetO gene. Five of the isolates were genetically unrelated as judged using pulsed field gel electrophoresis (PFGE) analysis. Two 19F isolates came from the same patient, carried both tetM and tetO genes and had the same PFGE pattern. The other 63 isolates carried only the tetM gene. DNA sequences from three of the tetO carrying isolates were determined; they showed 91-95% nucleotide sequence identity over 300 nucleotides, and 93-95% amino acid sequence identity over 100 amino acids. The isolates carrying both tetO and tetM genes could transfer the tetM gene into both Enterococcus faecalis and S. pneumoniae recipients, but not the tetO gene. There was no detectable transfer of the tetO gene, by conjugation, from the other five isolates. PMID- 9848446 TI - Relationship between antimycobacterial activities of rifampicin, rifabutin and KRM-1648 and rpoB mutations of Mycobacterium tuberculosis. AB - We compared the in-vitro antimycobacterial activities of rifabutin and KRM-1648, two rifamycin derivatives, with that of rifampicin against 163 strains of Mycobacterium tuberculosis. We also evaluated the correlation between the level of resistance to rifampicin, rifabutin and KRM-1648 and genetic alterations in the rpoB gene. All 82 strains susceptible to rifampicin or resistant to rifampicin with MICs < or = 16 mg/L were susceptible to rifabutin and KRM-1648 with MICs < or = 1 mg/L. Seventy-six of 81 strains resistant to rifampicin with MICs > or = 32 mg/L were resistant to both rifabutin and KRM-1648, but with lower MICs than those of rifampicin. KRM-1648 showed more potent antimycobacterial activity than rifabutin against organisms with low MICs (< or = 1 mg/L), while rifabutin was more active than KRM-1648 against organisms with high MICs (> or = 2 mg/L). A total of 96 genetic alterations around the 69 bp core region of the rpoB gene were detected in 92 strains. Alterations at codons 515, 521 and 533 in the rpoB gene did not influence the susceptibility to rifampicin, rifabutin and KRM-1648. Point mutations at codons 516 and 529, deletion at codon 518 and insertion at codon 514 influenced the susceptibility to rifampicin but not that to rifabutin or KRM-1648. With the exception of one strain, all alterations at codon 513 and 531 correlated with resistance to the three test drugs. The resistant phenotype of strains with an alteration at codon 526 depended on the type of amino acid substitution. Our results suggest that analysis of genetic alterations in the rpoB gene might be useful not only for predicting rifampicin susceptibility, but also for deciding when to use rifabutin for treating tuberculosis. Further studies may be required to determine the usefulness of KRM 1648. PMID- 9848447 TI - Effects of sucrose and silver on Staphylococcus aureus biofilms. AB - The present study examined the effects of antimicrobial agents on a biofilm model of Staphylococcus aureus. A membranous structure (an immature biofilm) was formed on the coverslips of tissue culture dishes by S. aureus cells in plasma after incubation for 24 h. After incubation, the colony counts of S. aureus cells in the immature biofilms in plasma containing levofloxacin with 70% sucrose were about one-tenth of those in plasma containing levofloxacin without sucrose. The colony counts of S. aureus cells in the immature biofilms in plasma containing silver sulphadiazine or silver nitrate (at a silver concentration of 0.302%) were over 3000 times lower than those in control plasma without silver after incubation for 24 h. Plasma coagulation by S. aureus cells was not detected in plasma containing 70% sucrose 48 h after inoculation. The immature biofilms were incubated in plasma with and without 70% sucrose, and the immature biofilms were treated with 10% povidone-iodine for 2 min after incubation for 4 and 16 h. In this experiment, the colony counts of S. aureus cells in the immature biofilms incubated with 70% sucrose were about one-tenth of those in plasma without sucrose after incubation for 24 h. We suggest that levofloxacin or 10% povidone iodine in combination with 70% sucrose and silver sulphadiazine or silver nitrate (at a silver concentration of 0.302%) are effective in eliminating S. aureus cells in a biofilm. PMID- 9848448 TI - Toxicity and therapeutic efficacy of amphotericin B delivered through cholesterol hemisuccinate vesicles in the treatment of experimental murine aspergillosis. AB - We have studied the toxicity and therapeutic efficacy of amphotericin B in cholesterol hemisuccinate vesicles (ABCV) in the treatment of invasive aspergillosis in Balb/c mice. The toxicity of amphotericin B was significantly reduced when delivered through cholesterol hemisuccinate vesicles as compared to deoxycholate suspension (AmB(DOC)) as evidenced by reduced nephrotoxicity in Balb/c mice, lower in-vitro toxicity to erythrocytes and higher maximum tolerated dose. The latter increased from 2 mg/kg wt for AmB(DOC) to 17 mg/kg wt for ABCV. The vesicles had a mean diameter of 252 nm. In order to evaluate the therapeutic efficacy of ABCV, Aspergillus fumigatus-infected mice were treated with ABCV 2, 4, 8 or 12 mg/kg or AmB(DOC) 1 mg/kg wt. The antifungal activity was highly dependent on the dosage of ABCV on the basis of survival of treated mice and cfu in lung, liver, spleen and kidney. This study found that ABCV had dose-dependent antifungal activity and therapeutic efficacy in the treatment of invasive aspergillosis in Balb/c mice. PMID- 9848449 TI - In-vitro activity of sanfetrinem against isolates of Streptococcus pneumoniae and Staphylococcus aureus. AB - The activity of sanfetrinem (previously GV104326), was assessed against 168 isolates of Streptococcus pneumoniae and 90 isolates of Staphylococcus aureus. These isolates included a range of serotypes or phage types, and varied in their susceptibility to other antibiotics. Sanfetrinem exhibited good anti-pneumococcal activity, with MIC90s of < or = 0.007 mg/L and 0.5 mg/L for penicillin susceptible and penicillin-resistant isolates, respectively. Sanfetrinem was also active against methicillin-susceptible staphylococci (MIC90 = 0.06 mg/L). However, the MICs of sanfetrinem for isolates with methicillin MICs of 8-16 mg/L and > or = 32 mg/L were 0.25-1 mg/L and 8->32 mg/L, respectively. PMID- 9848450 TI - In-vitro antimicrobial activity of HMR 3004 (RU 64004) against erythromycin A sensitive and -resistant Corynebacterium spp. isolated from clinical specimens. AB - We studied the in-vitro activity of HMR 3004 (RU 64004), a new ketolide, against 161 clinical isolates of Corynebacterium spp. including isolates resistant to erythromycin A, josamycin and lincomycin. HMR 3004 was active against all erythromycin A-sensitive isolates as well as against 75.8% and 45.4% of erythromycin A-intermediate and -resistant isolates, respectively. In contrast, HMR 3004 was active against 40 (46.5%) of 86 isolates resistant to erythromycin A, josamycin and lincomycin as well as against two isolates that were resistant to erythromycin A and lincomycin but not resistant (i.e. susceptible or intermediate) to josamycin. PMID- 9848452 TI - In-vitro evaluation of nitrofurantoin as an alternative agent for metronidazole in combination antimicrobial therapy against Helicobacter pylori. AB - Increasing metronidazole resistance suggests the need for alternative antibiotics for combination therapy of Helicobacter pylori infections. We evaluated a metronidazole-resistant and a clarithromycin-resistant strain of H. pylori under stationary growth phase conditions that favoured physiological conditions in order to determine if nitrofurantoin might be a suitable alternative for metronidazole in combination therapy. The results demonstrated that the triple combination of bismuth, tetracycline and nitrofurantoin achieved greater bactericidal activity against these two strains than did the combination of bismuth, tetracycline and metronidazole. These results suggest that further evaluation is warranted. PMID- 9848451 TI - Susceptibility to RPR 106,972, quinupristin/dalfopristin and erythromycin among recent clinical isolates of enterococci, staphylococci and streptococci from North American medical centres. AB - An orally administered streptogramin (RPR 106,972) and a parenteral streptogramin (quinupristin/dalfopristin) were evaluated against a collection of 2481 recent clinical isolates of gram-positive cocci. The isolates were gathered from ten North American medical centres during the winter months of 1996-1997. In spite of minor differences, both streptogramins had essentially identical spectra of activity which included many erythromycin-resistant isolates. Previously proposed interpretative criteria for quinupristin/dalfopristin disc diffusion susceptibility tests were confirmed. PMID- 9848453 TI - Absence of a post-antibiotic effect (PAE) of beta-lactams against Helicobacter pylori NCTC 11637. AB - A continuous culture system (chemostat) was used to study the post-antibiotic effect (PAE) of beta-lactams, against slowly-growing Helicobacter pylori NCTC 11637. H. pylori was grown at one quarter of its maximum specific growth rate (mu(max)) before exposure to ampicillin, amoxycillin, azlocillin, piperacillin or cefixime (8 x MIC). After 8 h, the antibiotics were inactivated. Viable counts were used to determine the rate of recovery of H. pylori. The recovery growth rate of H. pylori was similar to the maximum growth rate of H. pylori under antibiotic-free experimental conditions, so none of the beta-lactams studied showed a PAE against slowly-growing H. pylori. PMID- 9848455 TI - Carbon dioxide requirements of Streptococcus pneumoniae and Haemophilus influenzae. PMID- 9848454 TI - Use of ketolides in combination with other drugs to treat experimental toxoplasmosis. AB - Because combination therapy is required to treat human toxoplasmosis, we examined combinations of the ketolides HMR 3004 and HMR 3647 with atovaquone, clindamycin or sulphadiazine in a murine model of toxoplasmosis. An oral dose of 50 mg/kg/day of HMR 3004 protected 30% of mice lethally infected with Toxoplasma gondii. The same dose protected 100% of infected mice when administered in combination with non-protective doses of atovaquone, clindamycin or sulphadiazine. Similar results were noted with 25 mg/kg/day of HMR 3647. These results demonstrate that these drug combinations are highly effective for treating toxoplasmosis in mice. PMID- 9848456 TI - Comparative in-vitro activity of levofloxacin against Chlamydia spp. PMID- 9848458 TI - Glycopeptide resistance amongst Staphylococcus spp. PMID- 9848457 TI - In-vitro activities of gatifloxacin, sparfloxacin and trovafloxacin against 103 strains of Legionella spp. PMID- 9848459 TI - High prevalence of ciprofloxacin resistance amongst strains of Neisseria gonorrhoeae isolated from commercial sex workers in Bangladesh. PMID- 9848460 TI - Antibiotic usage and methicillin-resistant Staphylococcus aureus: an analysis of causality. PMID- 9848461 TI - Can a single antibiotic policy for the empirical treatment of febrile neutropenic patients be used for all categories of haematology/oncology patient in the same institution? PMID- 9848462 TI - Factors and their origins in mate selection and choice among humans: implications for individual psychotherapy and counseling. AB - A mechanism of mate selection in humans is proposed and elaborated. It is further proposed that this mechanism constitutes one of the important factors for stability and the necessary longevity of the procreational dyad and therefore the procreational success of humans as a species. The concepts and mechanisms of assortative mating (homogamy) and that of complementarity of temperaments of the mates (heterogamy) which guide such selections are described, the relationships between the two are explored, and finally their possible early developmental origins are proposed. Evidence from a small study of 20 married couples' responses in temperament tests is offered as well as some illustrative case histories all pointing to those mechanisms. The argument is based mainly on principles of evolutionary psychology. PMID- 9848463 TI - Fractal organization of the pointwise correlation dimension of the heart rate. AB - OBJECTIVE: To depict and quantify the degree of organization of the heart rate variability (HRV) in normal subjects. METHODS: A modified algorithm was created to estimate series of 'point-dimensions' (PD2) from interbeat (R-R) interval series of 10 healthy subjects (21-56 years). Our innovation is twofold: (i) we quantified instances of low-dimensional chaos, random fluctuations, and those for which our method failed to provide either (due to poor statistics); (ii) consecutive subepochs of PD2s underwent a relative dispersion (RD) analysis, yielding an index (D) which quantifies the dynamical organization of the heart rate generator. RESULTS: The mean values of PD2 series varied between 4.58 and 5.88 (mean+/-SD= 5.21+/-0.41, n = 10). For group 1 (21-30 years, n = 6) we found an averaged PD2 of 5.49+/-0.27, while for group 2 (47-56 years, n = 4) PD2 averaged 4.79+/-0.17. The RD analysis performed for subepochs of PD2s yielded both instances obeying fractal scaling (D < 1.5) and stochasticity (D > 1.5). The average D for group 1 was 1.39+/-0.04 (14 subepochs) and for group 2, 1.20+/ 0.008 (8 subepochs). Paired t-test and Hartley F-max test for comparison between D values and homogeneity of variance between the two groups were performed, yielding P-values 0.004 and 0.02, respectively. CONCLUSIONS: The complexity of the HRV seems to be modulated by a non-random fractal mechanism of a 'hyperchaotic' system, i.e. it can be hypothesized to contain more than one attractor. Also, our results support the 'chaos hypothesis' put forth recently, namely, the complexity of the cardiovascular dynamics is reduced with aging. The index of relative dispersion of the dimensional complexity has to be tested in various clinico-pathological settings, in order to corroborate its value as a potential new physiological measure. PMID- 9848464 TI - Does the lack of the P-glycoprotein efflux pump in neutrophils explain the efficacy of colchicine in familial Mediterranean fever and other inflammatory diseases? AB - Colchicine is an alkaloid drug commonly used in familial Mediterranean fever (FMF), gout, Behcet's syndrome, psoriasis and Sweet's syndrome. The exact mechanism of its action in these diseases is not entirely known. However, it has been shown that colchicine may inhibit neutrophil chemotaxis, thereby decreasing the inflammatory process. Recently, it was shown that colchicine accumulates in neutrophils in higher concentrations than in lymphomonocytes. Studies dealing with the multiple drug resistance (MDR) issue disclosed that neutrophils lack the P-glycoprotein (P-gly) membranal pump (encoded by the MDR1 gene). We propose that the preferential accumulation of colchicine in neutrophils compared with lymphomonocytes is due to the absence of the P-gly efflux pump in the former. This may explain the effectiveness of colchicine in diseases where increased chemotaxis is evident. The hypothesis may also provide an explanation for FMF patients who do not respond to the drug. PMID- 9848466 TI - Insights into the relationship of fatty streaks to raised atherosclerotic lesions provided by the hemorheologic-hemodynamic theory of atherosclerotic lesions provided by the hemorheologic-hemodynamic theory of atherogenesis. AB - The hemorheologic-hemodynamic theory of atherogenesis suggests that atherosclerosis is a disease of low shear, which prolongs the residence time of atherogenic particles on the endothelium. Prolonged residence of lipid-rich particles results in a fatty streak. Prolonged residence of platelet microthrombi results in a raised lesion (atherosclerotic plaque). Thus, fatty streak and raised lesion development are independent processes. In contrast, received wisdom holds that fatty streaks are the precursors to raised lesions. The author examines anatomic and risk factor data for fatty streaks and raised lesions, including the results of the recent multicenter Pathobiological Determinants of Atherosclerosis in Youth study, in light of these two theories. PMID- 9848465 TI - The treatment of spinocerebellar ataxias: facts and hypotheses. AB - Actual therapeutic assays in spinocerebellar ataxias, i.e. in Friedreich's ataxia (FA) and olivopontocerebellar atrophy (OPCA) are discussed in relation to (i) the serotoninergic theory; (ii) the excitotoxic action of glutamate; and (iii) cerebrospinal fluid thiamine deficiency in ataxic patients. Data from the literature show that neurochemical deficiencies arising from cerebellar damage in both FA and OPCA patients are multiple. Assays of replacement and neuroprotective therapeutics with a single drug have produced controversial data or mildly effective results. Consequently, it is hypothesized that a drug cocktail, i.e. L 5-hydroxytryptophan, thiamine and amantadine hydrochloride, would be more beneficial. This cocktail proved to be useful in open studies, improving respiratory disorders in FA patients. More powerful inhibitors of N-methyl-D aspartate receptor channels should be tried initially in animal experiments. PMID- 9848467 TI - The inter-relation of maternal immune competence, HIV-1 viral load, and nutritional status in preventing vertical transmission: an alternative to chemoprophylaxis? AB - As the human immunodeficiency virus (HIV) global pandemic moves towards the end of its second decade, women of reproductive age throughout the world have been shown to be increasingly at risk for acquiring HIV-1 infection. Recently, the focus for preventive measures has expanded to include preventing the perinatal transmission of HIV-1 to fetuses and newborns. This manuscript reviews the available literature that examines risk factors for perinatal transmission, immunopathogenesis of HIV-1 infection, and the role that antioxidant micronutrients play in modulating immune response to HIV-1 disease progression. The available information provides a compelling case for the design of studies that evaluate the extent to which maternal HIV-1 viremia and disease progression are modulated by her nutritional status. Should results from these studies confirm that antioxidant micronutrient status is inversely related to HIV-1 RNA load, particularly in economically vulnerable populations, carefully designed and executed supplementation trials would be warranted. PMID- 9848468 TI - Utility of metformin as an adjunct to hydroxycitrate/carnitine for reducing body fat in diabetics. AB - Excessive exposure of tissues to fatty acids is likely to be the chief cause of the various dysfunctions that lead to sustained hyperglycemia in type II diabetes. These dysfunctions are likely to be substantially reversible if body fat and dietary fat can be greatly reduced. Disinhibition of hepatic fatty acid oxidation with hydroxycitrate (HCA) and carnitine has considerable potential as a new weight-loss strategy, but in diabetics runs the risk of further enhancing excessive hepatic gluconeogenesis. Since the clinical utility of metformin in diabetes is probably traceable to inhibition of gluconeogenesis, its use as an adjunct to HCA/carnitine treatment of obesity in diabetics deserves evaluation, particularly as metformin therapy itself tends to reduce body weight. A consideration of relevant evidence suggests that metformin therapy will not impede the activation of fatty acid oxidation by HCA/carnitine, and is likely to potentiate the appetite-suppressant and thermogenic benefits of this strategy. Indeed, since metformin has been reported to lower body weight and improve cardiovascular risk factors in obese non-diabetics, a broader application of a metformin/HCA/carnitine therapy for obesity can be contemplated. PMID- 9848469 TI - Antioxidant inhibitors for cancer therapy. AB - Built-in cellular defense mechanisms play a major role in a tumor's protection against non-surgical antineoplastic therapies. Of these, the overexpression of antioxidants such as superoxide dismutase (SOD) may be the most important. Oxygen radicals are highly toxic, and have been implicated in various diseases, including carcinogenesis and aging. They produce a variety of pathological changes through lipid peroxidation and DNA damage. Therefore, treating free radical-induced diseases with antioxidants has been an accepted therapeutic approach. Ironically, however, the underlying mechanism that most chemotherapeutic agents and ionizing radiation exert on tumor cell kill is not increased antioxidation but rather the production of more free radicals leading to irreversible tissue injury. A small increase in reactive oxygen species (ROS) following non-surgical antineoplastic therapies induces the expression of antioxidants such as SOD, but overproduction of ROS, conversely, exhausts the production of SOD and other adaptive antioxidant defenses. Based on these considerations, we hypothesize that the appropriate administration of antioxidant inhibitors and/or free-radical-generating compounds may be a useful strategy in the treatment of solid tumors. PMID- 9848470 TI - Host metabolism: a target in clinical oncology? AB - The presence of the tumour induces important metabolic changes in the cancer patient which are not merely due to the fact that the tumour acts as a parasite, thus depleting the host of nutrients, but that are mainly the result of both tumoral and humoral mediators. The new metabolic status of the cancer patient may lead to cancer cachexia (a pathological state characterized by weight loss together with anorexia, weakness, anaemia and asthenia) which represents one of the worst effects of malignancy, accounting for nearly a third of cancer deaths. The complications associated with the appearance of the cachectic syndrome affect both the physiological and biochemical balance of the patient and have effects on the efficiency of the anticancer treatment, resulting in a considerably decreased survival time. At the metabolic level, cachexia is associated with loss of skeletal muscle protein together with a depletion of body lipid stores. The present study emphasizes the fact that neutralizing some of the metabolic changes in the patient may be an essential therapeutic strategy in controlling tumour growth and improving survival. PMID- 9848471 TI - Facilitating survival and recovery by removing autogenic pathology from the clinical picture. AB - For the most part, the clinical picture is the expression of autogenic pathology and, unless the anatomical insult represents a major structural compromise, it contributes little to the clinical picture. Autogenic pathology created by the processing of peripheral neurological impulses altered by the presence of anatomical insults is created independently of the gravity of the anatomical insults. Even in the presence of a survivable insult, autogenic pathology is often fatal. Each successful resuscitation effort identifies a patient with a survivable insult who almost died before autogenic pathology was removed by cerebral anoxia, and/or electric shock. Similar patients who suffer terminal brain damage cannot be identified. Cardinal receptors furnish the embryo with its first neural contact with its environment. On completion of embryological migration, they occupy a position in the skin overlying the root of their spinal nerve. Cardinal receptors are functionally segmented to the specific distribution of their spinal nerve and use the stable neurological impulses from the skin to furnish the cadence by which peripheral impulses are processed and valued. Selected cardinal receptors may be specifically stimulated by saline infiltration that produces intradermal blebs in the skin overlying their spinal nerve's root. An augmented burst of infiltration pain immediately followed by symptomatic relief identifies monitoring cardinal receptors. The identification of monitoring receptors confirms the presence of pathology found within the specific distribution of their spinal nerve. Peripheral impulses processed within the cadence of highly stimulated cardinal receptors are assigned an insignificant value. Assigning an insignificant value to peripheral impulses altered by the presence of an anatomical insult removes autogenic pathology from the clinical picture. The immediate onset of profound symptomatic relief heralds the removal of autogenic pathology. Visceral pathology is monitored by bilateral pairs of cardinal receptors so it produces discomfort perceived without specific location information. Therefore, the specific visceral distribution has been discounted and is virtually unknown. However, when visceral pathology is correlated with bilateral pairs of monitoring cardinal receptors there are no patient variations. PMID- 9848472 TI - Microbiology of the intestinal lymph follicle: a clue to elucidate causative microbial agent(s) in Crohn's disease. AB - It has been suggested that microbial agent(s) are involved in the onset of Crohn's disease. None of the candidates, however, has been unequivocally demonstrated to be a causative agent. The macroscopically earliest lesion takes place in the lymph follicle, irrespective of the initial attack or relapse in Crohn's disease. Human leucocyte antigen-DR (HLA-DR) antigens are expressed on the epithelium around the lymph follicle even in areas endoscopically uninvolved in Crohn's disease. These observations make the lymph follicle critical in the onset of Crohn's disease. The lymph follicle is a port of entry of a variety of microbial agent(s), leading to the speculation that microbial agent(s) exist in the lymph follicle. Polymerase chain reaction (PCR) using universal primers designed from conserved regions of bacterial ribosomal RNA or techniques such as representational difference analysis, may well identify microbial agent(s) in the lymph follicle that are specific to Crohn's disease. The existence of bacteria in the lymph follicle is here indicated by preliminary studies. PMID- 9848473 TI - Syndrome-AC: non-insulin-dependent diabetes mellitus and the anabolic/catabolic paradox. AB - When an organism is starving, infected or injured, the body initiates a catabolic response that, among other things, creates insulin resistance. While many possible mechanisms and numerous loci of insulin resistance have been studied, a theme of inappropriately activated components of catabolic chemistry emerges. Patients with non-insulin-dependent diabetes mellitus (NIDDM) have elevated blood levels of glucagon, cortisol, fatty acids, protein, glucose and possibly acute phase reactants. Recent studies hint that this can occur with meals, resulting in a multilevel, multiorgan interference with glucose handling. This implies NIDDM is the result of the dietary activation of catabolic chemistry simultaneously with that of anabolic chemistry. We review the possibility that this is caused by the consumption of body tissue, the substance ordinarily endogenously released in starvation or injury. Activating the catabolic pathways when eating creates a hormonal paradox, forcing exaggerated insulin levels to compensate. Five case studies are reviewed. PMID- 9848474 TI - A possible neurophysiological basis for psychological pain. AB - There is some evidence that the anterior cingulate cortex (ACC) contains nociceptive neurons, and other neurons which respond to conditioned aversive stimuli. It is not implausible that nociceptive input and conditioned aversive input converge onto common neurons in the ACC. Following from this hypothesis, the psychological pain which occurs in circumstances that arouse disgust or aversion may involve the same mechanisms that give rise to referred pain. PMID- 9848475 TI - Extrapineal melatonin and exogenous serotonin in seasonal affective disorder. AB - Visible light inhibits the binding of melatonin and serotonin to cultured human peripheral blood mononuclear leukocytes (PBMLs) in winter. The decreased binding switches the metabolism in PBMLs towards serotonin synthesis, resulting in the reduced production of melatonin. The ingestion of L-tryptophan during the day is hypothesized to increase the levels of melatonin, released from the gastrointestinal tract, in patients with winter seasonal affective disorder (SAD). Due to the relative shortage of light, coincident with a predisposed metabolic error, there would be no switch towards serotonin synthesis among winter SAD patients in winter. The rate of serotonin synthesis could thus remain inadequately low to maintain optimal mood in winter SAD patients. PMID- 9848476 TI - The amount of DNA in cells of the granular layer of the cerebellum and their susceptibility to hypoxia. AB - Purkinje cells of the cerebellum are particularly susceptible to hypoxia. In these cells tetraploidy has been demonstrated. Therefore, a link between the susceptibility of cells of the cerebellum to hypoxia and the amount of DNA seems probable. PMID- 9848477 TI - On the possibility of preventing multiple sclerosis. AB - The role of immune complexes in the etiology and pathogenesis of demyelinating diseases, particularly multiple sclerosis, is discussed in the light of numerous observations. It is inferred that a deeper understanding of the role of immune complexes might provide a basis for preventive measures in multiple sclerosis. PMID- 9848478 TI - Magnetoreception attributed to the efficacy of light therapy. AB - A group of specialized photoreceptors are suggested to amplify the weak interaction of the geomagnetic field with a single electron spin to the level of photon detection, resulting in a modulation of the response to light. Under exposure to rotating magnetic field, the size of pinealocytes is bigger at night than during the day in spring, but there is no day-night difference in autumn. Specialized photoreceptors are hypothesized to modulate the response of the photoreceptive system to light, and the pineal response to a magnetic stimulus, in patients with winter seasonal affective disorder (SAD). This could explain why some winter SAD patients show excessive sensitivity to light in winter. PMID- 9848479 TI - Moral dilemmas of tetraplegia; the 'locked-in' syndrome, the persistent vegetative state and brain death. PMID- 9848480 TI - Maintenance of locomotor abilities following Laufband (treadmill) therapy in para and tetraplegic persons: follow-up studies. AB - Recent reports indicate that walking capabilities in spinal cord damaged persons significantly improve--as compared to conventional rehabilitation therapy--after intensive training of aided (Laufband) treadmill-stepping. In the present report, follow up investigations on two collectives of spinal cord injured (sci) persons are described who had undergone (Laufband) treadmill therapy either during a period of renewed rehabilitation months or years after spinal cord injury (35 chronic patients) or during their first postacute rehabilitation period (41 acute patients). Among the initially chronic patients, 20 from 25 still wheelchair bound before the onset of (Laufband) treadmill therapy, ie not capable of raising from the wheelchair and walking without help by other persons, became independent walkers after therapy. Assessment of voluntary muscle activity in resting position before and after the period of therapy had shown only small increases in most patients, indicating the involvement of motor automatisms and better utilisation of remaining muscle function during walking. Follow-up assessments performed 6 months to 6 1/2 years after discharge from the hospital revealed that the walking capabilities achieved by (Laufband) treadmill therapy in the 35 initially chronic patients were maintained in 31 persons, in three they had further improved, in only one it was reduced. These results indicate that the improvements achieved under clinical conditions can be maintained in every day life under domestic surroundings. From 41 initially acute patients, 15 had further improved and none had reduced his walking capability 6 months to 6 years after discharge from the hospital. PMID- 9848481 TI - Augmented upper body contribution to oxygen uptake during upper body exercise with concurrent leg functional electrical stimulation in persons with spinal cord injury. AB - The purpose of this pilot study was to compare the contribution of upper body musculature to VO2 with and without concurrent leg FES (LFES). Eight subjects with spinal cord injury, lesion levels range C6-T12, performed upper body exercise (UBE) during no LFES (NOS), LFES at 40 mA (LOS), and 80 mA (HIS), at rest, 60% and 80% of VO2peak. Resting VO2 values were obtained during NOS, LOS and HIS conditions and were then subtracted from their respective whole body VO2 values to give an estimate of upper body VO2. Small and non significant increases were found in the HIS vs NOS condition at 60% VO2peak. Larger differences of 7.8% were found in the HIS vs NOS condition at 80% VO2peak (11.35+/-3.8 ml kg(-1) min( 1) to 12.24+/-4.0 ml kg(-1) min(-1)), although this too was not significant, perhaps due to the small number of subjects in this study and the consequently low statistical power to detect a significant difference. We discuss the implications for these preliminary results in the context of the existing literature on this topic. PMID- 9848482 TI - Autonomic dysreflexia during urodynamics. AB - Autonomic dysreflexia (AD) is an acute syndrome characterised by inappropriate and massive autonomic response that occurs in patients with spinal cord injury above the T6 level. AIMS: to evaluate the incidence of AD during cystometry and the relationships with clinical and urodynamic features. PATIENTS AND METHODS: Forty-eight spinal cord injury patients were studied by neurological and urological examination and urodynamic evaluation with concurrent recording of blood pressure, heart rate and symptoms and signs of AD. Patients were considered to have AD if blood pressure reached values higher than 150/100 mmHg. RESULTS: All the patients showed a significant increase of both systolic and diastolic blood pressure, although only 20 showed pressure values higher than 150/100 mmHg (in seven of them without AD symptoms). AD was more frequent in cervical patients (P = 0.034), but did not correlate with any other clinical features: sex ratio, age, disease duration, completeness of lesion, incidence of detrusor hyperreflexia/areflexia and detrusor-sphincter dyssynergia, voiding modalities, usage of anticholinergic drugs. In three patients blood pressure increase began when uninhibited contraction started, in 11 it was coincident with uninhibited contraction peak and in the other six it appeared at maximum bladder capacity. CONCLUSIONS: (1) during urodynamic evaluation all the patients with lesion level above T6 showed signs of sympathetic stimulation, although only some showed dangerous blood pressure values; (2) the relationship between urodynamic data and dysreflexia crisis shows that both the presence of detrusor uninhibited contractions and bladder distension are able to stimulate the crisis; (3) treatment with anticholinergic drugs is not sufficient to prevent autonomic dysreflexia starting from the bladder, unless it induces detrusor areflexia. These patients are at risk of developing autonomic dysreflexia following bladder distension. PMID- 9848483 TI - Pathophysiology of autonomic dysreflexia: long-term treatment with terazosin in adult and paediatric spinal cord injury patients manifesting recurrent dysreflexic episodes. AB - INTRODUCTION: Spinal cord injury (SCI) results in disruption of synaptic influences on the sympathetic preganglionic neurones. Remodelling of spinal cord circuits takes place in spinal neurones caudal to cord injury. There is an increased vascular alpha-adrenoceptor responsiveness, and peripheral afferent (bladder) stimulation in SCI subjects induces a marked noradrenaline spillover below the level of spinal lesion. These neurophysiological changes possibly contribute to the development of autonomic dysreflexia, a condition of sympathetic hyper-excitability that develops after cervical, or upper dorsal cord injury with resultant paroxysmal rise in arterial pressure, and provide the scientific basis for the use of terazosin, a once-a-day, selective alpha-one adrenergic blocking drug. OBJECTIVES: The use of terazosin, a long-acting, alpha 1-selective blocking agent was investigated in SCI patients who developed recurrent symptoms of autonomic dysreflexia, eg headache, sweating flushing of the face together with an increase in the arterial pressure. DESIGN: An open, prospective study of the efficacy of terazosin in controlling recurrent autonomic dysreflexia in traumatic tetraplegic/paraplegic patients manifesting clinical features of dysreflexia in the absence of an acute precipitating cause such as a blocked catheter. SETTING: The initial assessment and treatment were carried out in the Spinal Injuries Centre. Subsequently, the patients were followed-up in the community. They were monitored by telephonic interviews, follow-up visits by the patients to the hospital, and home-visits by the staff of the spinal unit. SUBJECTS: Eighteen adults with tetraplegia (female: 1; male: 17), three children with ventilator-dependent tetraplegia and three adult male patients with paraplegia who exhibited recurrent features of autonomic dysreflexia in the absence of an acute predisposing factor for dysreflexia eg performance of an invasive procedure such as cystoscopy, digital evacuation of bowels, or acute urinary retention, were enrolled in this study. INTERVENTION: After discussion with the patients and their carers, terazosin was prescribed with a starting dose of 1 mg in an adult and 0.5 mg in a child administered nocte. The patients were observed for (1) drug-induced hypotension; (2) clinical symptoms due to side effects of terazosin; and (3) continued occurrence of dysreflexic symptoms. Step wise increments of the dose of terazosin (1 mg in case of adults, and 0.5 mg in a child) was carried out at intervals of 3-4 days, if a patient continued to develop dysreflexia but did not manifest any serious side effect. OUTCOME MEASURES: Complete subsidence of dysreflexic symptoms, or development of an adverse event necessitating termination of the terazosin therapy was the clinical end point. RESULTS: The dysreflexic symptoms subsided completely with the terazosin therapy in all the patients. The twenty-one adult patients required a dose varying from 1-10 mg, whereas the paediatric patients required only 1-2 mg of terazosin. The side effects of postural hypotension and drowsiness were transient, and mild. One tetraplegic patient developed persistent dizziness and therefore, the drug therapy was discontinued. CONCLUSION: In 21 adult and three paediatric spinal cord injury patients manifesting recurrent episodes of autonomic dysreflexia in the absence of an acute predisposing cause, the use of terazosin, a once-a-day, specific alpha-one blocker resulted in complete subsidence of the dysreflexic symptoms. However, one tetraplegic patient required termination of terazosin therapy because of persistent dizziness. PMID- 9848484 TI - Seminal plasma PSA in spinal cord injured men: a preliminary report. AB - OBJECTIVE: The aim of the study was to evaluate the impact of spinal cord injury on seminal plasma PSA concentration. MATERIAL: Semen obtained from normal controls (n = 20) and patients (n = 16) with spinal cord injury. RESULTS: The concentration of prostate specific antigen in seminal plasma depended on the level of spinal cord injury, with significantly lower concentrations in patients with spinal cord lesions below T7 compared to both normal age-matched controls (P = 0.007) and patients with spinal cord injuries at or above T7 (P = 0.017). CONCLUSIONS: The results add to the previously reported observations of the possible impact of neurogenic stimulation on prostate activity. More studies of neurogenic stimulation in prostate growth and development are warranted to further elucidate the possible impact of neurogenic factors in the development of prostatic diseases. PMID- 9848485 TI - Faecal incontinence in children with spina bifida: the best conservative treatment. AB - Incontinence is one of the major stigmata affecting the quality of life of the myelodysplastic patient. The appropriate management of the neuropathic bladder has led most patients to survive with good conditions. Major advances have also been made in the treatment of faecal incontinence for enabling patients to achieve their full potential, to become integrated in society and to be as independent as possible. We present the results of the Retrograde Colonic Enema (RCE) in 33 patients suffering neurogenic faecal incontinence. We believe that the RCE is one of the best conservative methods of treatment for relieving faecal incontinence originating from spina bifida and other neurological problems with intestinal dysfunction. PMID- 9848486 TI - Polyethylene glycol versus vegetable oil based bisacodyl suppositories to initiate side-lying bowel care: a clinical trial in persons with spinal cord injury. AB - INTRODUCTION: Neurogenic bowel dysfunction resulting from spinal cord injury (SCI) frequently requires bowel care (BC) with stimulant suppositories for initiation of effective defecation. The excessive time required for BC and bowel complications have limited quality of life after SCI. OBJECTIVE: To test the hypothesis that: the time required for bowel care with bisacodyl suppositories can be reduced by substituting a polyethylene glycol base (PGB) for the traditional hydrogenated vegetable oil base (HVB) in the suppository. SETTING: Inpatient SCI medicine unit. SUBJECTS: Fourteen persons with SCI with chronic stable paralysis from upper motor neuron SCI for greater than one year with a stable HVB bisacodyl suppository initiated BC. DESIGN: Crossover Controlled. METHOD: Subjects received HVB bisacodyl suppositories for six sequential BC sessions and then were crossed over to PGB bisacodyl suppositories for six more BCs. OUTCOME MEASURES: BC event times were utilized to derive BC intervals: suppository insertion to first flatus= Time to flatus, first flatus until the beginning of stool flow = Flatus to stool flow, begin stool flow until end stool flow = Defecation period, end stool flow until end of clean up = Clean up, and suppository insertion until end clean up = Total bowel care time. RESULTS: The data included two groups of BC sessions: HVB (n = 84) and PGB (n = 81). Mean times in minutes and P values from t tests for paired samples yielded: Time to flatus: (HVB 31, PGB 12.8 P < 0.002), Defecation period: (HVB 58, PGB 32, P < 0.0005), Clean up: (HVB 1.9, PGB 3.2 P = 0.165), Total bowel care time: (HVB 102, PGB 51.2 P < 0.0005). CONCLUSION: This analysis suggests that PGB based bisacodyl suppositories may stimulate reflex defecation sooner and shorten the Total BC Time as compared with HVB bisacodyl suppositories. PMID- 9848487 TI - Detraining from total body exercise ergometry in individuals with spinal cord injury. AB - This study was designed to investigate the effects of detraining that occurred during an 8 week period of muscular inactivity following a 12 week training program of artificial computerized functional electrical stimulation cycle ergometry (CFES LE) and arm ergometry. Six spinal cord injured male individuals were followed through an 8 week detraining period that was preceded by a 12 week exercise program including CFES LE and arm ergometry. Maximal graded exercise tests were completed and measurements of peak oxygen consumption (VO2), heart rate (HR), ventilation (VE) workload, and creatine kinase were taken. Testing occurred at initial training (0T), after 12 weeks of training (12T), and after 8 weeks of detraining (DT). After the training program, peak VO2 increased significantly from 0.562 +/- 0.126 (0T) to 1.021 +/- 0.247 l/min (12T, P < 0.05). After DT, peak VO2 decreased to 0.791 +/- 0.216 l/min, which was lower than 12T (P < 0.05), yet higher than 0T (P < 0.05). After DT, peak workoad had decreased from 0.675 +/- 0.203 (12T) to 0.32 +/- 0.203 kp (P < 0.05), which was not different than 0T. Creatine kinase levels were significantly lower both at 12T and DT compared to 0T (P < 0.05). In addition, this training program induced linear increases in both VO2 and HR with workload, which were retained after DT. These increases did not reach statistical significance. however. No apparent relationship existed between these values at baseline. There were no significant differences in submaximal or peak HR of VE between the three testing periods. The results indicate that both peripheral muscular adaptations and central distribution adaptations in SCI individuals are partially maintained following 8 weeks of DT from CFES LE and arm ergometry. PMID- 9848488 TI - Fracture rates and risk factors for fractures in patients with spinal cord injury. AB - AIM: To study fracture rates and risk factors for fractures in patients with spinal cord injuries. MATERIAL AND METHODS: A self-administered questionnaire was mailed to 646 members of the Danish Paraplegic Association and 1000 randomly selected normal controls. 438 patients (309 males, 129 females, 67.8%) and 654 controls (332 males, 322 females, 65.4%) returned the questionnaire. Median age in patients was 42, range 10-80 years, and in controls 43, range 19-93 years (2p = 0.25). RESULTS: The crude fracture rate was 2% per year in patients and 1% per year in controls (RR = 2.0, P < 0.001). Low-energy fractures were much more prominent in patients (19.0% of all fractures) than in controls (1.4%, P < 0.001). The fracture rate did not differ before the injury but increased after the injury to a constant level from the third year and forward. Fractures of the lower extremities were more prominent in patients than controls (femurs: RR = 23.4, P < 0.001, lower legs: RR = 5.2, P < 0.001, feet/toes: RR = 2.4, P = 0.006) while fractures of the forearms (P < 0.001) and clavicles (P = 0.03) were absent among patients. Fractures were more frequent in female patients (RR = 1.6, P = 0.008) and in male patients with a family history of fractures (RR = 2.0, P = 0.004). CONCLUSIONS: Low-energy fractures especially of the lower extremities are frequent in spinal cord injury patients and especially among female patients. The forearms seem protected from fractures. PMID- 9848489 TI - Brown-Sequard syndrome caused by a Kirschner wire as a complication of clavicular osteosynthesis. AB - A case of spinal cord injury caused by delayed migration of a Kirschner wire is reported. Some cases of distant injuries caused by bone wires, and acupuncture needles have been published, but this is the first reported case of delayed thoracic spinal cord damage caused by the migration of a clavicular wire. A 22 year-old male patient was admitted with a clinical picture of spinal shock after performing physiotherapeutic exercises. Two months prior to this, the patient had undergone surgical treatment for a clavicular fracture in a different clinical center. Imaging showed a clavicular wire had migrated into the spinal canal. An early prescription of a spinal cord methyl-prednisolone protective treatment (NASCIS II), the surgical extraction of the foreign body and the rehabilitation exercises were the keys to a quick recovery. PMID- 9848490 TI - Aspergillus spondylodiscitis in an immunocompetent paraplegic patient. AB - A case of an immunocompetent 60 year old patient is reported, who suffered extensive thoracic spinal injury and paraplegia after polytrauma. In the course of rehabilitation he developed aspergillus spondylodiscitis in a part of the thoraco-lumbar spine which was primarily uninjured. The diagnostic assessment and therapeutic approach of this rare disorder is elucidated and discussed in the context of paraplegia and polytrauma. Possible mechanisms of inoculation and spreading of the moulds as well as predisposing factors of the disease are discussed in this paper and a review of the recent literature is provided. PMID- 9848492 TI - Life expectancy in traumatic spinal injuries. PMID- 9848493 TI - Prolonged penile erection association with terazosin in a cervical spinal cord injury patient. PMID- 9848494 TI - The Role of Soy in Preventing and Treating Chronic Disease. Proceedings of a symposium. Brussels, Belgium, September 15-19, 1996. PMID- 9848491 TI - 'Camel collision as a major cause of low cervical spinal cord injury'. Spinal Cord (1998) 36, 415-417. PMID- 9848495 TI - Biotechnology and the soybean. AB - Glyphosate-tolerant soybeans (GTSs), the first biotechnologically improved soybeans to be marketed, became available commercially in 1996. The safety of GTSs was assessed in 2 ways: study of the introduced protein and of the soybean seed and selected processing fractions. Because soybeans are a major source of protein in most farm animals' diets, animal feeding studies in wholesomeness were done to complement the analyses. Analysis of the expressed protein in GTSs [3 phosphoshikimate 1-carboxyvinyltransferase (EC 2.5.1.19)] showed it to be readily digestible and to possess no allergenic concerns. In addition, comparison of the composition of seeds and selected processing fractions from 2 GTS lines with the parental line showed that the lines are equivalent. Feeding studies in various animal species confirmed that the feeding value of GTS lines is comparable with that of the parental line. These studies support the conclusion that GTSs are as safe and nutritious as traditional soybeans marketed currently and can be incorporated safely into feed and food products in the near future. PMID- 9848496 TI - Phytoestrogens: the biochemistry, physiology, and implications for human health of soy isoflavones. AB - The importance of estrogens in homeostatic regulation of many cellular and biochemical events is well illustrated by the pathophysiologic changes that occur with estrogen deficiency. Many of the major diseases of Western populations are hormone dependent and epidemiologic data have shown a strong association between their incidence and diet. In particular, the importance of a plant-based diet is evident from the current dietary recommendations that emphasize an increase in the proportion and amount of fruit and vegetables that should be consumed. Although interpretation of the role of individual components of the diet is difficult from epidemiologic and dietary studies, it is recognized that there are many plant-derived bioactive nonnutrients that can confer significant health benefits. Among these phytochemicals is the broad class of nonsteroidal estrogens called phytoestrogens, and in the past decade there has been considerable interest in the role of isoflavones because of their relatively high concentrations in soy protein. The isoflavones in modest amounts of ingested soy protein are biotransformed by intestinal microflora, are absorbed, undergo enterohepatic recycling, and reach circulating concentrations that exceed by several orders of magnitude the amounts of endogenous estrogens. These phytoestrogens and their metabolites have many potent hormonal and nonhormonal activities that may explain some of the biological effects of diets rich in phytoestrogens. PMID- 9848497 TI - Effects of soy protein on renal function and proteinuria in patients with type 2 diabetes. AB - For > 150 y, clinicians and investigators have observed that high protein intakes accelerate the progression of renal disease and that low protein intakes have beneficial effects. Some studies suggest that the effects of soy-protein intake resemble those of a low-protein diet. The Brenner hypothesis suggests that high protein intakes by diabetic individuals create hyperfiltration and glomerular hypertension eventuating in renal damage. On the basis of the available evidence, we are proposing the soy-protein hypothesis, which states that substituting soy protein for animal protein in diabetes patients results in less hyperfiltration and glomerular hypertension and, therefore, resultant protection from diabetic nephropathy. Furthermore, substituting soy protein for animal protein should have therapeutic value in diabetic nephropathy with resultant slowing of deterioration of renal function and decreasing proteinuria. The preliminary results of the study of 8 type 2 diabetes patients with obesity, hypertension, and proteinuria are reported. Under the conditions of the study, providing soy protein as half of the daily protein intake had no distinct effects on renal function or proteinuria in these subjects. Soy-protein intake was associated with a significant reduction in serum cholesterol and triacylglycerol concentrations. Further studies are required to critically examine the effects of soy-protein intake on the renal function of diabetes patients. PMID- 9848498 TI - Soy isoflavonoids exhibit in vitro biological activities of loop diuretics. AB - Intake of soyfoods containing isoflavonoids is related to positive effects on heart and kidney diseases. Urinary equol, a potent inhibitor of Na+-K+-2Cl- cotransport, originates from the metabolism of daidzein by intestinal bacteria. Loop diuretics (eg, furosemide), acting through inhibition of Na+-K+-2Cl- cotransport, are used to maintain adequate blood volume. In the present work, we compare isoflavonoids' inhibition of cotransport and effects on the function and hemodynamics of isolated perfused rat kidneys with those of furosemide. Equol [IC50 (half-maximal inhibitory concentration): 23.6 +/- 3.6 micromol/L], genistein (IC50: 34.8 +/- 2.6 micromol/L), and daidzein (IC50: 140.0 +/- 24 micromol/L) inhibited bumetanide-sensitive rubidium uptake in LLC-PK1 cells. The IC50 of equol and genistein was close to that of furosemide (IC50: 10.3 +/- 2.7 micromol/L). Furosemide, equol, and genistein stimulated water, sodium, and potassium excretion by isolated rat kidneys in the same temporal pattern. None of the isoflavonoids significantly increased the glomerular filtration rate, but genistein induced significant vasorelaxation. We conclude that isoflavonoids exhibit biological activities of furosemide in vitro, at concentrations similar to those reported for other in vitro effects. More research is needed to evaluate the participation of cotransport inhibition by isoflavonoids in the healthful effects claimed for soy intake. PMID- 9848499 TI - Role of soy protein with normal or reduced isoflavone content in reversing bone loss induced by ovarian hormone deficiency in rats. AB - Soy protein, a rich source of isoflavones, fed immediately after an ovariectomy prevents bone loss in rats. Reports of the effectiveness of natural and synthetic isoflavones in preventing or treating osteoporosis led us to examine the effect of soy protein in reversing established bone loss. Seventy-two 95-d-old female Sprague-Dawley rats were assigned to 6 groups. The rats were either sham operated (SHAM; 2 groups) or ovariectomized (OVX; 4 groups) and then fed a casein-based, semipurified diet. Thirty-five days after surgery, 1 SHAM and 1 OVX group were killed to examine the occurrence of bone loss. Thereafter, the other SHAM and 1 OVX groups continued to receive the casein-based diet. Whereas the remaining 2 OVX groups received diets in which casein was replaced by soy protein with normal (OVX+SOY) or reduced (OVX+SOY-) isoflavone content for 65 days. The OVX control group had significantly lower femoral and fourth lumbar vertebral bone densities than the SHAM group. Femoral density of rats fed SOY or SOY- diets were not significantly different from SHAM or OVX controls. This suggests a slight reversal of cortical bone loss that may be partially due to higher femoral insulin-like growth factor I mRNA transcripts resulting from both the SOY and SOY diets. The ovariectomy-induced increases in indexes of bone turnover were not ameliorated by either of the soy diets, suggesting that any positive effect of soy was achieved through enhanced bone formation rather than slowed bone resorption. Long-term consumption of soy or its isoflavones may be needed to produce small but continued increments in bone mass. PMID- 9848500 TI - Bone-sparing effect of soy protein in ovarian hormone-deficient rats is related to its isoflavone content. AB - Our previous studies showed that a soy-protein diet prevents ovariectomy-induced bone loss. The purpose of this study was to determine whether isoflavones in soy protein are responsible for this bone-protective effect. Forty-eight 95-d-old Sprague-Dawley rats were divided into 4 groups: sham-operated fed a casein-based diet (SHAM), ovariectomized fed a casein-based diet (OVX+CASEIN), ovariectomized fed soy protein with normal isoflavone content (OVX+SOY), and ovariectomized fed soy protein with reduced isoflavone content (OVX+SOY-). The OVX+SOY group had significantly greater femoral bone density (in g/cm3 bone vol) than the OVX+CASEIN group, whereas OVX+SOY- was similar to OVX+CASEIN (mean +/- SD; SHAM, 1.522 +/- 0.041; OVX+CASEIN, 1.449 +/- 0.044; OVX+SOY, 1.497 +/- 0.030; OVX+SOY-, 1.452 +/- 0.030). Ovariectomy resulted in greater bone turnover as indicated by higher serum alkaline phosphatase activity, serum insulin-like growth factor I and insulin-like growth factor binding protein 3 concentrations, and urinary hydroxyproline. These increases were not affected by soy with either normal or reduced isoflavone content. Similarly, histomorphometry revealed a greater bone formation rate with ovariectomy, and this was not altered by the soy diets. The findings of this study suggest that isoflavones in soy protein are responsible for its bone-sparing effects. Further studies to evaluate the mechanism of action of isoflavones on bone are warranted. PMID- 9848501 TI - Tyrosine kinase inhibitor effects on avian osteoclastic acid transport. AB - We found that tyrosine kinase pp60(c-src) coisolates with acid-transporting osteoclast membranes and hypothesized that this kinase regulates hydrochloric acid transport. We assayed the membrane acid transport and bone degradation effects of tyrosine kinase inhibitors in avian osteoclasts. Isoflavone, tyrphostin, and benzoquinonoid inhibitors were compared with inactive analogues to determine nonspecific effects. Acid-secreting membranes, isolated by nitrogen cavitation, were assayed as reconstituted vesicles by using acridine orange to indicate ATP-dependent hydrogen ion transport. The soy isoflavone genistein and the benzoquinonoid antibiotic herbimycin inhibited hydrochloric acid transport with 50% inhibition at approximately 10 and approximately 2 micromol/L, respectively; effects appeared in <2 min and were reversible. In membrane incubated with inhibitors, the herbimycin effect also inhibited Cl- transport by variable amounts, suggesting that this compound affects Cl- channel activity. However, genistein and tyrphostins did not produce chloride dependent effects. After 30 min with ATP, tyrphostin A47 irreversibly inhibited hydrochloric acid transport with 50% inhibition at approximately 10 micromol/L. Tyrphostin A25 and controls, tyrphostin A1 and daidzein (a genistein congener), were inactive despite preincubation. Osteoclastic bone resorption was more sensitive to the inhibitors over 3-5-d assays than was membrane acid transport, except for tyrphostins. Herbimycin and genistein inhibited bone resorption with half maximal effects at 0.5 and 10 micromol/L and complete inhibition at 3 d in 1 and 20 micromol/L, respectively. None of the tyrphostins, including A47, nor daidzein inhibited resorption to >20 micromol/L. We conclude that tyrosine kinase inhibition directly inhibits osteoclast membrane hydrochloric acid transport; differences among inhibitors may reflect chemical reactivity and permeability. PMID- 9848502 TI - Soy protein and isoflavones: their effects on blood lipids and bone density in postmenopausal women. AB - The effects of soy protein (40 g/d) containing moderate and higher concentrations of isoflavones on blood lipid profiles, mononuclear cell LDL receptor messenger RNA, and bone mineral density and content were investigated in 66 free-living, hypercholesterolemic, postmenopausal women during a 6-mo, parallel-group, double blind trial with 3 interventions. After a control period of 14 d, during which subjects followed a National Cholesterol Education Program Step I low-fat, low cholesterol diet, all subjects were randomly assigned to 1 of 3 dietary groups: Step I diet with 40 g protein/d obtained from casein and nonfat dry milk (CNFDM), Step I diet with 40 g protein/d from isolated soy protein containing 1.39 mg isoflavones/g protein (ISP56), or Step I diet with 40 g protein/d from isolated soy protein containing 2.25 mg isoflavones/g protein (ISP90). Total and regional bone mineral content and density were assessed. Non-HDL cholesterol for both ISP56 and ISP90 groups was reduced compared with the CNFDM group (P < 0.05). HDL cholesterol increased in both ISP56 and ISP90 groups (P < 0.05). Mononuclear cell LDL receptor mRNA was increased in subjects consuming ISP56 or ISP90 compared with those consuming CNFDM (P < 0.05). Significant increases occurred in both bone mineral content and density in the lumbar spine but not elsewhere for the ISP90 group compared with the control group (P < 0.05). Intake of soy protein at both isoflavone concentrations for 6 mo may decrease the risk factors associated with cardiovascular disease in postmenopausal women. However, only the higher isoflavone-containing product protected against spinal bone loss. PMID- 9848503 TI - Variable lipemic response to dietary soy protein in healthy, normolipemic men. AB - We found previously that dietary soy protein, compared with casein, reduced plasma LDL cholesterol and increased HDL cholesterol concentrations in healthy women and men. However, there was considerable variation among individuals. The aim of this study was to characterize the lipoprotein responsiveness of individuals to examine whether different response patterns could be identified. Nine normolipemic men consumed 2 liquid-formula diets of identical composition except that the protein component was either soy protein or casein. After 1 mo of consuming each diet, the subjects' plasma HDL cholesterol (P < 0.01) and apolipoprotein (apo) A-I (P < 0.05) concentrations were increased by the soy protein diet whereas the ratio of LDL cholesterol to HDL cholesterol was decreased (P < 0.01); total cholesterol, triacylglycerol, LDL cholesterol, apo B and apo A-II were insignificantly affected. In 5 individuals, however, soy protein reduced mean LDL cholesterol, LDL2 cholesterol, and LDL2 apo B concentrations by 26% and plasma apo B by 16%, whereas HDL cholesterol increased by 11%. In 3 other individuals, soy protein increased mean HDL cholesterol by 17% and plasma apo A-I by 12%, but did not lower LDL. In 1 subject, soy protein decreased LDL2 cholesterol by 11% and increased plasma triacylglycerol by 40%, but neither HDL cholesterol nor apo A-I increased. We identified 3 types of lipemic responses to dietary soy protein involving a reduction in atherogenic LDL and increase in antiatherogenic HDL. In most subjects, the effects on both LDL and HDL were favorable, although fewer experienced either an increase in HDL or a decrease in LDL2. PMID- 9848504 TI - Cholesterol-lowering effect of soy protein in normocholesterolemic and hypercholesterolemic men. AB - Cardiovascular heart disease is a major health problem in the United States. Elevated blood cholesterol has been shown to significantly increase the risk of cardiovascular heart disease. The National Cholesterol Educational Program (NCEP) Step I diet, which restricts fat and cholesterol intakes, is usually recommended as the initial treatment to lower blood cholesterol. Soy protein has been shown to be hypocholesterolemic, particularly in hypercholesterolemic subjects. However, the hypocholesterolemic effect of soy protein in subjects with a blood total cholesterol concentration <5.17 mmol/L is not clear. To determine whether soy protein could enhance the hypocholesterolemic effect of the NCEP Step I diet, 13 normocholesterolemic and 13 hypercholesterolemic men aged 20-50 y were enrolled in a randomized, 2-part, crossover study. Subjects were fed either an NCEP Step I soy-protein diet or an NCEP Step I animal protein diet for 5 wk. After a washout period of 10-15 wk, the subjects were fed the alternate diet for 5 wk. The hypocholesterolemic effect of soy protein was found to be independent of age, body weight, pretreatment plasma lipid concentrations, and sequence of dietary treatment. Regardless of plasma lipid status, the soy-protein diet was associated with a statistically significant decrease in the plasma concentrations of LDL cholesterol (P = 0.029) as well as the in the ratio of plasma LDL cholesterol to HDL cholesterol (P = 0.005). Our results indicate that soy protein enhances the hypocholesterolemic effect of the NCEP Step I diet in both normocholesterolemic and hypercholesterolemic men. PMID- 9848505 TI - Effects of soy isoflavones on atherosclerosis: potential mechanisms. AB - It has long been recognized that coronary heart disease rates are lower in Japan, where soy consumption is common, than in Western countries. In experimental studies, atherosclerosis was reduced in animals fed diets containing soy protein compared with those fed diets with animal protein. Recently, several lines of evidence have suggested that the components of soy protein that lower lipid concentrations are extractable by alcohol (eg, the isoflavones genistein and daidzein). We recently evaluated the relative effect of the soy protein versus the alcohol-extractable components of soy on cardiovascular disease and its risk factors. Young male and female cynomolgus monkeys were fed diets that contained either 1) casein-lactalbumin as the source of protein (casein), 2) soy protein isolate from which the isoflavones were alcohol extracted (SPI-), or 3) isoflavone-intact soy protein (SPI+). The SPI+ group had significant improvements in LDL cholesterol and HDL cholesterol. Only HDL cholesterol was significantly improved in the SPI- group males compared with the casein group. The casein group had the most atherosclerosis, the SPI+ group had the least, and the SPI- group was intermediate but did not differ significantly from the casein group. Potential mechanisms by which soy isoflavones might prevent atherosclerosis include a beneficial effect on plasma lipid concentrations, antioxidant effects, antiproliferative and antimigratory effects on smooth muscle cells, effects on thrombus formation, and maintenance of normal vascular reactivity. PMID- 9848506 TI - Prevention of precancerous colonic lesions in rats by soy flakes, soy flour, genistein, and calcium. AB - The main purpose of this research was to determine whether diets containing soy products would inhibit the early stages of azoxymethane-induced colon cancer in F344 rats. Additional objectives were to determine whether feeding starch instead of sucrose, feeding additional calcium (0.5% compared with 0.1%), or feeding a low-fiber powdered enteral formula would influence early colon carcinogenesis. Colon cancer was initiated with 2 injections of azoxymethane (15 mg/kg body wt) and a 12-wk dietary treatment period was started 1 wk after the second injection. Precancerous colon lesions were assessed as foci with aberrant crypts (FAC). The mean numbers of FAC were 133 [soy concentrate (low concentration of phytochemicals)], 111 (starch substituted for sucrose), 98 [full-fat soy flakes (whole soybeans)], 87 (defatted soy flour), 77 (0.015% genistein), and 70 (0.5% Ca). The soy flour and full-fat soy flake diets contained 0.049% genistein derivatives (primarily glycosides), but were less effective in inhibiting the formation of FAC than the diet containing 0.015% genistein (as the aglycone). Eating soybeans and soy flour may reduce the early stages of colon cancer. PMID- 9848507 TI - Genistein studies in rats: potential for breast cancer prevention and reproductive and developmental toxicity. AB - Asian women and men who consume a traditional diet high in soy products have low incidences of breast and prostate cancers, respectively. Yet Asians who immigrate to the United States and adopt a Western diet lose this protection. We investigated the potential of genistein, a component of soy, to protect against breast cancer and to cause reproductive and developmental toxicity. Our study showed that injections of genistein in rats during the prepubertal period resulted in a 50% reduction of chemically induced mammary tumorigenesis. Studies in mammary whole mounts revealed that prepubertal genistein exposure resulted in fewer terminal end buds and more lobules type II. Cell proliferation in the terminal end buds of adult rats treated prepubertally with genistein was less than that in animals treated with the vehicle (dimethyl sulfoxide). Reproductive and developmental toxicity studies did not find significant alterations to fertility, number of male and female offspring, body weight, anogenital distance, vaginal opening, testes descent, estrus cycle, or follicular development. We concluded that pharmacologic doses of genistein given to immature rats enhance mammary gland differentiation, resulting in a significantly less proliferative gland that is not as susceptible to mammary cancer. We speculate that breast cancer protection in Asian women consuming traditional soy-containing diets is, in part, derived from early exposure to genistein-containing soy. We believe that early programming events are essential for cancer protection benefits. PMID- 9848508 TI - The Bowman-Birk inhibitor from soybeans as an anticarcinogenic agent. AB - Certain protease inhibitors are effective at preventing or suppressing carcinogen induced transformation in vitro and carcinogenesis in animal model systems. One protease inhibitor, the soybean-derived Bowman-Birk inhibitor (BBI) is particularly effective in suppressing carcinogenesis. BBI is a protein of a molecular weight of 8000 with a well-characterized ability to inhibit trypsin and chymotrypsin. BBI has been extensively studied, both as purified BBI and as an extract of soybeans enriched in BBI called BBI concentrate (BBIC). Purified BBI and BBIC have comparable suppressive effects on the carcinogenic process in a variety of in vivo and in vitro systems. BBI appears to be a universal cancer preventive agent. Purified BBI and BBIC suppress carcinogenesis as follows: in 3 different species (mice, rats, and hamsters); in several organ systems and tissue types [eg, colon, liver, lung, esophagus, cheek pouch (oral epithelium), and cells of hematopoietic origin]; and in cells of epithelial and connective tissue origin when given to animals by several different routes of administration, including the diet, leading to different types of cancer (eg, squamous cell carcinomas, adenocarcinomas, and angiosarcomas), and induced by various chemical and physical carcinogens. About half of an oral dose of BBI is taken up into the bloodstream and distributed throughout the body, with excretion via the urine. Pharmacokinetic studies of BBI have been performed in animals with radioactively labeled BBI, whereas antibodies that react with reduced BBI are being used in pharmacokinetic studies in humans. The calculated serum half-life is 10 h in both rats and hamsters. BBIC achieved Investigational New Drug status from the FDA in April 1992 (IND no. 34671; sponsor, Ann R Kennedy), and studies to evaluate BBIC as an anticarcinogenic agent in human populations began. Both BBI and BBIC prevent and suppress malignant transformation in vitro and carcinogenesis in vivo without toxicity. PMID- 9848509 TI - Effects of mammalian and plant estrogens on mammary glands and uteri of macaques. AB - Because of their possible beneficial effects on atherosclerosis and cancer risk, soy-derived isoflavone phytoestrogens may be useful as a dietary alternative or supplement to postmenopausal hormone replacement therapy. We examined this possibility in a well-characterized primate model of postmenopause. Adult, surgically postmenopausal female macaques (Macaca fascicularis) were treated continuously with either estradiol (E2), an isoflavone-rich soy protein isolate (SPI), or both (E2+SPI). Doses were equivalent on an energy basis to 1 and 148 mg/d per woman for E2 and SPI, respectively. After 6 mo, histopathologic, morphometric, and immunohistochemical measurements of the endometrium and mammary glands were taken. Increases in endometrial thickness, gland area, and epithelial proliferation were induced by E2 and E2+SPI. Morphometric changes were accompanied by increased epithelial staining of the proliferation marker Ki-67 in the E2-treated group. The effects of E2 were partially antagonized by SPI (manifested as decreased Ki-67 staining). Mammary gland proliferation was induced by E2 and E2+SPI. The effects of E2 were also antagonized by SPI in the mammary gland. Morphometric and immunohistochemical measures of proliferation were in agreement in endometrium. In this nonhuman primate model, treatment with SPI did not induce proliferation in endometrial and mammary tissue. SPI may have antiproliferative effects in the endometrium and mammary gland when given along with exogenous estrogen. PMID- 9848510 TI - Mechanisms of action of the soy isoflavone genistein: emerging role for its effects via transforming growth factor beta signaling pathways. AB - The soy isoflavone genistein attenuates growth factor- and cytokine-stimulated proliferation of both normal and cancer cells. This article reviews our current understanding of the potential mechanisms of action of genistein. In membrane preparations from mammalian cells, genistein is a potent and specific inhibitor of tyrosine autophosphorylation of the epidermal growth factor (EGF) receptor. However, in several cell systems in which it inhibits growth, genistein does not alter tyrosine phosphorylation of the EGF receptor or other tyrosine kinase substrates thought to be involved in signal transduction pathways, suggesting that other mechanisms may be responsible for its action. Alternatives include inhibition of DNA topoisomerase II activity, regulation of cell cycle checkpoints, and antiangiogenic and antioxidant activity. Experiments in our laboratory suggest a new concept, that genistein may inhibit cell growth by modulating transforming growth factor (TGF) beta1 signaling pathways. Such a link between genistein action and TGFbeta1 function is supported by preliminary results of studies in patients with hereditary hemorrhagic telangiectasia (a genetic disorder involving mutations in proteins that regulate TGFbeta receptor complex formation and signaling) in which several patients had dramatic attenuation of their symptoms after 1 wk of ingesting soy-based beverages. These preclinical studies in combination with our cell culture data suggest that the mechanism of genistein involves, if not requires, TGFbeta1-signaling. PMID- 9848511 TI - Genistein induces maturation of cultured human breast cancer cells and prevents tumor growth in nude mice. AB - Results of recent studies in animal models of mammary carcinogenesis showed that the soybean isoflavone genistein is a chemopreventive agent. The objective of the present study was to determine whether soybean isoflavones can be used for the prevention of human breast carcinogenesis. Human adenocarcinoma cells that are either estrogen-receptor positive (such as MCF-7) or estrogen-receptor negative (such as MDA-MB-468) were used as our model system. Treatment of these cells with genistein concentrations of 15, 30, and 45 micromol/L resulted in cell growth inhibition, which was accompanied by the expression of maturation markers. Maturation was monitored by the induction of intracytoplasmic casein and lipids and the membrane protein intercellular adhesion molecule-1. These maturation markers were optimally expressed after 9 d of treatment with 30 mmol genistein/L. Both estrogen receptor-positive and -negative cells became differentiated in response to genistein treatments, suggesting that the antiestrogenic function of genistein is unrelated to the mechanism of cell differentiation. Daidzein, the other major isoflavone component of soybeans, did not induce differentiation in either MCF-7 or MDA-MB-468 cells. To explore the potential applications of this result, we used the nude mouse xenograft model of carcinogenesis. Treatment of either cell line with genistein before implantation into nude mice diminished the cells' tumorigenic potential. These data suggest that initiation of the differentiation program provides a protective effect against tumor growth in mouse xenografts. PMID- 9848512 TI - Effects of soy-protein supplementation on epithelial proliferation in the histologically normal human breast. AB - A high dietary intake of soy products (eg, as in Japan and Singapore) has been associated with a reduction in the incidence of breast cancer in premenopausal women. Phytoestrogens present in soybeans inhibit human breast cancer cell proliferation in vitro and breast cancer development in animal models, but no data exist on the effects of phytoestrogens on histologically normal human breasts. This study examines the effects of dietary soy supplementation on the proliferation rate of premenopausal, histologically normal breast epithelium and the expression of progesterone receptor. Women (n = 48) with benign or malignant breast disease were randomly assigned to receive their normal diet either alone or with a 60-g soy supplement (containing 45 mg isoflavones) taken daily for 14 d. Biopsy samples of normal breasts were labeled with [3H]thymidine to detect the number of cells in S phase and were immunocytochemically stained for the proliferation antigen Ki67. The phytoestrogens genistein, daidzein, equol, enterolactone, and enterodiol were measured in serum samples obtained before and after supplementation. Serum concentrations of the isoflavones genistein and daidzein increased in the soy group at 14 d. Results showed a strong correlation between Ki67 and the thymidine labeling index (r = 0.868, P < or = 0.001). The proliferation rate of breast lobular epithelium significantly increased after 14 d of soy supplementation when both the day of menstrual cycle and the age of patient were accounted for. Progesterone receptor expression increased significantly in the soy group. Short-term dietary soy stimulates breast proliferation; further studies are required to determine whether this is due to estrogen agonist activity and to examine the long-term effects of soy supplementation on the pituitary gland and breast. PMID- 9848513 TI - Soy intake and risk of breast cancer in Asians and Asian Americans. AB - Evidence from case-control studies suggests, although not entirely consistently, that soy intake may protect against breast cancer. The designs and findings of studies conducted in Asian women living in Japan, Singapore, China, and the United States are reviewed. Because of the considerably higher intake of soy by native Asians than by Asian Americans living in California and Hawaii, these studies investigated different segments of the dose-response relation between soy intake and breast cancer risk. Data are not sufficient to determine the amount or frequency of soy intake effective in protecting against breast cancer. Of concern is that soy intake may be homogeneously high in Asia, making it difficult to identify differences in breast cancer risk between high and moderate daily consumers. In studies conducted in Asian Americans, it is difficult to be certain that soy intake is not a marker of other factors related to Western lifestyle that are causally associated with risk of breast cancer. Additional studies assessing the role of soy and breast cancer are needed. These studies should assess intake of all food sources of soy, considering portion size as well as other dietary and nondietary factors that may confound the soy-breast cancer association. A better understanding of the mechanisms whereby soy intake may influence the risk of breast cancer is also needed. Dietary intervention studies with soy will provide information on the acute effects of soy on endogenous hormone concentrations. Cross-sectional and longitudinal studies are necessary to investigate the longer-term relations between hormone concentrations and soy intake in women. PMID- 9848514 TI - Use of soy-protein formulas and soyfood for feeding infants and children in Asia. AB - Soybeans have been cultivated and consumed in Asia for many centuries. Soy products can be found in all households in Asian countries, and Asian children begin to consume soy formulas and soy products at a very young age. In a study of soy exposure in a group of healthy Singaporean children < 10 y of age, 70% had consumed soy products and of those > 95% had consumed soy products before the age of 18 mo. Soy products are commonly used as food flavorings and for weaning Asian children with lactose intolerance or allergy to cow-milk protein. The widespread use of soy formulas and soy products by Asian children is mainly due to the high nutritive value and palatability of these products. PMID- 9848515 TI - Soy protein for the prevention and treatment of children with cow-milk allergy. AB - Soy-protein formulas are widely used for feeding babies with cow-milk allergy. When they first were marketed, these formulas were the only available cow-milk substitute and they ensured a normal life for many children who were affected by the large spectrum of clinical manifestations of cow-milk allergy. Soy-protein formulas were also given to allergy-prone infants for the prevention of atopic diseases when breast milk was not available. Several researchers studied the prevalence of soy sensitization in allergic disease. Few studies used a challenge test for the diagnosis of soy allergy, even those in patients in whom soy allergy was suspected. In most studies the diagnosis of soy allergy was based on anecdotal case histories reported by parents and was not substantiated by scientific diagnostic criteria: no challenge test to soy was made nor were data available on specific immunoglobulin E to soy. In this paper we critically reviewed literature on the safety of feeding soy-protein formulas to babies with cow-milk allergy as well as on the prevention of cow-milk allergy. PMID- 9848516 TI - Isoflavone content of infant formulas and the metabolic fate of these phytoestrogens in early life. AB - Soy-based infant formulas have been in use for >30 y. These formulas are manufactured from soy protein isolates and contain significant amounts of phytoestrogens of the isoflavone class. As determined by HPLC, the isoflavone compositions of commercially available formulas are similar qualitatively and quantitatively and are consistent with the isoflavone composition of soy protein isolates. Genistein, found predominantly in the form of glycosidic conjugates, accounts for >65% of the isoflavones in soy-based formulas. Total isoflavone concentrations of soy-based formulas prepared for infant feeding range from 32 to 47 mg/L, whereas isoflavone concentrations in human breast milk are only 5.6 +/- 4.4 microg/L (mean +/- SD, n = 9). Infants fed soy-based formulas are therefore exposed to 22-45 mg isoflavones/d (6-11 mg x kg body wt(-1) x d(-1)), whereas the intake of these phytoestrogens from human milk is negligible (<0.01 mg/d). The metabolic fate of isoflavones from soy-based infant formula is described. Plasma isoflavone concentrations reported previously for 4-mo-old infants fed soy-based formula were 654-1775 microg/L (mean: 979.7 microg/L: Lancet 1997:350;23-7), significantly higher than plasma concentrations of infants fed either cow-milk formula (mean +/- SD: 9.4 +/- 1.2 microg/L) or human breast milk (4.7 +/- 1.3 microg/L). The high steady state plasma concentration of isoflavones in infants fed soy-based formula is explained by reduced intestinal biotransformation, as evidenced by low or undetectable concentrations of equol and other metabolites, and is maintained by constant daily exposure from frequent feeding. Isoflavones circulate at concentrations that are 13,000-22,000-fold higher than plasma estradiol concentrations in early life. Exposure to these phytoestrogens early in life may have long-term health benefits for hormone-dependent diseases. PMID- 9848518 TI - Isoflavones in human breast milk and other biological fluids. AB - We established a method for using HPLC and diode-array ultraviolet scanning to quantitate soy isoflavonoids in foods and in human plasma, urine, and breast milk. The analytes occurring as glycoside conjugates were hydrolyzed enzymatically before HPLC analysis if extracted from biological matrices or were subjected to direct HPLC analysis after extraction from foods. We monitored the isoflavones daidzein, genistein, glycitein, formononetin, and biochanin-A and their mammalian metabolites equol and O-desmethylangolensin in human plasma, urine, and breast milk. Analytes were identified by absorbance patterns, fluorometric and electrochemical detection. and comparison with internal and external standards. In addition, we identified analytes by using gas chromatography-mass spectrometry after trimethylsilylation. The HPLC method was also used to measure concentrations of isoflavones and their glucoside conjugates in various soy-based infant formulas. Total isoflavone concentrations varied between 155 and 281 mg/kg. After one woman received a moderate challenge with 20 g roasted soybeans (equivalent to 37 mg isoflavones), we detected mean total isoflavone concentrations of approximately 2.0 micromol/L in plasma, 0.2 micromol/L in breast milk, and 3.0 micromol/h in urine. According to our measurements, with adjustment for body weight, isoflavonoid exposure is 4-6 times higher in infants fed soy-based formula than in adults eating a diet rich in soyfoods (approximately 30 g/d). Implications of the presented results for the potential cancer-preventing activity of isoflavones by exposing newborn infants to these phytochemicals are discussed. PMID- 9848517 TI - Daily intake and urinary excretion of genistein and daidzein by infants fed soy- or dairy-based infant formulas. AB - Our aims were to measure isoflavone intake from soy- and dairy-based infant formulas and breast milk and to assess the ability of infants to digest and absorb soy isoflavones by measuring daily urinary excretion rates. We recruited 29 infants: 4 received soy-based formula and 25 received dairy-based formula. We collected pooled urine samples from 3-5 disposable diapers worn during a 24-h period and developed and validated methods for extracting isoflavones from the diapers. Infants were studied every 1 or 2 wk, starting at 2-6 wk of age and continuing until 16 wk. Only soy-based formulas contained isoflavones in concentrations detectable by HPLC (limits: 0.05 mg/L for liquids and 0.1 mg/kg for solids). Soy-based formulas provided a mean (+/-SEM) daily dose of isoflavones (genistein plus daidzein) of 3.2 +/- 0.2 mg/kg body wt, which remained fairly constant (CV: 12%) regardless of age < or = 16 wk. Isoflavones were measurable in all samples from soy-fed infants, but not in urine from dairy fed infants. Daily isoflavone excretion rates varied little among infants [range of mean individual values (mg x kg(-1) d(-1)): daidzein, 0.37 +/- 0.03 to 0.58 +/ 0.06; genistein, 0.15 +/- 0.03 to 0.32 +/- 0.04] and did not change with age < or = 16 wk. The mean percentage of the daily intake recovered in the urine of soy fed infants was 38 +/- 4% for daidzein and 13 +/- 3% for genistein, and remained constant with age. These values are similar to those for adults and indicate that young infants are able to digest, absorb, and excrete genistein and daidzein from soy-based formulas as efficiently as do adults consuming soy products. PMID- 9848519 TI - Soy isoflavone analysis: quality control and a new internal standard. AB - Development of a database of the soy isoflavone content of foods requires accurate and precise evaluation of different food matrixes. To evaluate accuracy, we estimated recoveries of both internal and external standards in 5 different soyfoods weekly. Standards were evaluated daily for system quality assurance. To evaluate sample precision, we analyzed soybeans and soymilk bimonthly for within day precision and over 4 d for day-to-day precision. CVs should be < or = 8%. We validated our methods for single and multiple recovery concentrations by using our new internal standard, 2,4,4'-trihydroxydeoxybenzoin, and the external standards daidzein, genistein, and genistin. Concentrations of 12 isoflavone isomers, 3 aglycones (daidzein, genistein, and glycitein), and 9 glucosides (daidzin, genistin, glycitin, acetyldaidzin, acetylgenistin, acetylglycitin, malonyldaidzin, malonylgenistin, and malonylglycitin) were measured in a variety of soybeans and soyfoods. The extraction methods used depended on soyfood type. The HPLC conditions for soy isoflavone analysis were improved, leading to good separation with a short analysis time (60 min/sample). A data bank of concentration and distribution of isoflavones in different soybean products was assembled. A wide range of isoflavone concentrations, from < 50 microg/g to > 20,000 microg/g, was found in different soy products. The glucoside forms are almost twice the molecular weight of the aglycones; reported isoflavone concentrations should be normalized to the aglycone mass (or an isoflavonoid equivalent) rather than a simple sum of all isomers. PMID- 9848520 TI - Quantification of isoflavones by capillary zone electrophoresis in soybean seeds: effects of variety and environment. AB - Soybean isoflavones (genistin, daidzin, glycitin, and their malonyl forms and aglucons) are thought to be responsible for the astringent taste of soyfoods. Generally, HPLC with a methanol and water elution gradient is used for isoflavone quantification, but capillary zone electrophoresis (CZE) has been used more recently to separate several flavonoids in plant extracts. We present the results of CZE analysis of isoflavones in soybean extracts. Conditions for separation by using CZE were optimized for analysis of soybean isoflavones. We compared the results of extraction at different temperatures and with different compositions of solvent. Total extraction of isoflavones was not affected by temperature but was affected by composition of the solvent. Malonyl forms of isoflavones were thermally unstable. We analyzed the isoflavone content of different varieties of soybean seeds sown on different dates. Total isoflavone content varied among different varieties and with sowing dates. Interactions between the variety and the sowing date also affected isoflavone composition. We conclude that the variety of soybean seed and environmental growing conditions, such as sowing date, can contribute to seed quality by reducing its isoflavone content, modifying its isoflavone composition, or both. PMID- 9848521 TI - Chemical modification of isoflavones in soyfoods during cooking and processing. AB - The principal chemical forms of isoflavones in soybean are their 6''-O-malonyl beta-glucoside (6OMalGlc) conjugates. Experiments were carried out to determine the best conditions for extraction of isoflavones from soyfoods and the effects of commercial processing procedures and of cooking on isoflavone concentrations and composition. Hot alcohol extraction of ground soybeans deesterified 6OMalGlc conjugates. Although room temperature extraction slowed the conversion, extraction at 4 degrees C for 2-4 h led to the highest yield of 6OMalGlc conjugates and the lowest proportion of beta-glucoside conjugates. Analysis of soyfood products by reversed-phase HPLC-mass spectrometry revealed that defatted soy flour that had not been heat treated consisted mostly of 6OMalGlc conjugates; in contrast, toasted soy flour contained large amounts of 6''-O-acetyl-beta glucoside conjugates, formed by heat-induced decarboxylation of the malonate group to acetate. Soymilk and tofu consisted almost entirely of beta-glucoside conjugates; low-fat versions of these products were markedly depleted in isoflavones. Alcohol-washed soy-protein concentrates contained few isoflavones. Isolated soy protein and textured vegetable protein consisted of a mixture of all 3 types of isoflavone conjugates. Baking or frying of textured vegetable protein at 190 degrees C and baking of soy flour in cookies did not alter total isoflavone content, but there was a steady increase in beta-glucoside conjugates at the expense of 6OMalGlc conjugates. The chemical form of isoflavones in foods should be taken into consideration when evaluating their availability for absorption from the diet. PMID- 9848522 TI - Influence of soybean processing, habitual diet, and soy dose on urinary isoflavonoid excretion. AB - In an attempt to explain the wide individual variation seen in urinary isoflavonoid phytoestrogen excretion, we conducted a series of 3 human feeding studies: a large cross-sectional study of equol production in humans with a soy challenge, a comparison of phytoestrogen metabolism when subjects consumed fermented and unfermented soy products, and a dose-response study of urinary isoflavonoid excretion at the low end of soy consumption. All studies were conducted in young, healthy humans. Urinary isoflavonoids were measured by isotope-dilution gas chromatography-mass spectrometry. Similar to results from other studies, 35% of screened subjects (30 men and 30 women) excreted equol (>2000 nmol/d). In women, equol excretion was associated with higher intake of dietary fiber and carbohydrate. Fermentation of soy decreased the isoflavone content of the product fed but increased the urinary isoflavonoid recovery, suggesting that fermentation increases availability of isoflavones in soy. When soy-protein powder was fed at 0, 5, 10, and 20 g/d (0-36 mg isoflavones), there was a linear dose response of urinary isoflavonoid excretion to soy consumption that did not differ between subjects with high and low equol excretion. These results suggest that equol excretion may be related to the fermentable carbohydrate content of the diet; additional study is needed. Processing of soy affects isoflavone metabolism and must be considered in recommending exposure to isoflavones from soyfoods. Although optimal isoflavone exposure for disease protection has not been determined, urinary isoflavonoid excretion appears linear at low-to-moderate soy consumption. PMID- 9848523 TI - Daidzein conjugates are more bioavailable than genistein conjugates in rats. AB - This study compared the bioavailability of conjugates of the soy isoflavones genistein and daidzein in rats. Rats were given a single oral dose of a soy extract that provided 74 micromol genistein and 77 micromol daidzein/kg body wt (as conjugates). Plasma samples were obtained from treated and untreated rats; urine and fecal samples were obtained before and after treatment. Isoflavones, equol (the main end product of bacterial degradation of daidzein), and 4-ethyl phenol (the main end product from genistein) were measured by HPLC. The plasma daidzein concentration was maximal at 2 h (9.5 +/- 0.71 micromol/L) and was almost double that of genistein (P = 0.009). Between 2 and 15 h, the plasma daidzein concentration declined by 32%, but the concentration of genistein changed little. At 15 h, the concentrations of daidzein and genistein were not significantly different. Urinary excretion of daidzein over the 48-h postdose period was 17.4 +/- 1.2% of the dose, but only 11.9 +/- 1.1% of the genistein dose was excreted in urine. Equol excretion was 5.0 +/- 1.5% of the daidzein dose, but 41.9 +/- 5.0% of the genistein dose was excreted as 4-ethyl phenol. Fecal daidzein accounted for 2.3 +/- 0.5% and fecal genistein for 3.4 +/- 0.4% of the respective doses. It is concluded that conjugates of daidzein are more bioavailable than those of genistein, probably because of the greater resistance of the former to degradation by gut bacteria. PMID- 9848524 TI - Sex and long-term soy diets affect the metabolism and excretion of soy isoflavones in humans. AB - Soybean consumption may be protective against hormone-dependent cancers, possibly in part because of the isoflavones daidzein and genistein, which are weakly estrogenic. This paper reviews our studies of the metabolism and disposition of these phytoestrogens in humans. During 1 mo of daily soy ingestion in a metabolic unit [1.065 L (36 oz) soymilk, providing 80-210 mg of each isoflavone daily], women initially excreted more isoflavone conjugates in urine than did men. Recoveries of conjugates of genistein, daidzein, and equol were 24%, 66%, and 28% of the amounts ingested in women, respectively, and 15%, 47%, and 15%, respectively, of those in men. A progressive decrease in urinary excretion of genistein and daidzein was observed in women but not in men during the study. At least 10% of ingested daidzin was excreted in urine as equol conjugate in one man and one woman after the first soy ingestion. Three more women but no more men developed the ability to produce and excrete large amounts of equol. Absorption rate constants (k(e)) of the isoflavones were estimated to be 0.24-0.50 h(-1). The elimination rates (k(e)) for genistein, daidzein, and equol were 0.1, 0.16, and 0.08 h(-1), respectively, in women and 0.19, 0.25, and 0.13 h(-1), respectively, in men. Thus, the excretion half-life values of genistein were longer in women (7, 4, and 9 h, respectively) than in men (4, 3, and 5 h, respectively) after the first soy ingestion. The excretion half-life shortened progressively in women but lengthened progressively in men over the study period. Thus, isoflavone metabolism and disposition were affected by the duration of soy ingestion and by sex. PMID- 9848525 TI - Metabolism of the isoflavones genistein and biochanin A in human breast cancer cell lines. AB - There is substantial variation in the growth inhibition of different human breast cancer cell lines by the isoflavones genistein and biochanin A. ZR-75-1 and BT-20 cells are > or = 2- to 4-fold less sensitive to these isoflavones than are MCF-7 cells, whereas T47D cells have a sensitivity similar to that of MCF-7 cells. To determine whether these differences are related to isoflavone metabolism by these cancer cells, each of the cell lines was incubated with [4-(14)C]genistein and [4 (14)C]biochanin A. Metabolites in the cell culture media were identified by radio HPLC electrospray ionization mass spectrometry. One metabolite of genistein (genistein 7-sulfate) and 2 metabolites of biochanin A (genistein and genistein 7 sulfate) were detected by radio-HPLC. Further analysis by mass spectrometry identified 3 other metabolites, a hydroxylated methylated form of each isoflavone and a biochanin A sulfate. IC50 (the concentration at which the growth rate was halved) values of the breast cancer cell lines did not correlate well with production of genistein 7-sulfate from genistein or with biochanin A sulfate, genistein 7-sulfate, or genistein from biochanin A. However, IC50 values correlated with the production of the hydroxylated and methylated forms of the isoflavones. Only T47D cells produced these metabolites in this study, and only T47D cells had IC50 values similar to those of MCF-7 cells, which also produced the hydroxylated and methylated metabolites. These data suggest that the hydroxylated and methylated metabolites may be the active forms of genistein in human breast cancer cells and emphasize the importance of isoflavone metabolism in the mechanism of action of isoflavones. PMID- 9848526 TI - Effects of isoflavones on alcohol pharmacokinetics and alcohol-drinking behavior in rats. AB - Puerarin, daidzin, and daidzein are 3 major isoflavonoid compounds isolated from Pueraria lobata, an edible vine used widely in China for various medicinal purposes. We studied the antiinebriation and the antidipsotropic effects of these antioxidants in rats. Daidzin and daidzein shortened alcohol-induced sleep time (loss of righting reflex) in rats that were given ethanol intragastrically but not in those given ethanol intraperitoneally. When daidzin was given to animals intragastrically with the ethanol solution, the blood alcohol concentration (BAC) was found to peak later and be lower than in control rats that were given only the ethanol solution. BACs also receded more slowly if daidzin was fed to the animals. None of the 3 isoflavonoid compounds administered orally affected liver alcohol dehydrogenase or aldehyde dehydrogenase activities, as was reported for intraperitoneal administration. Further experiments indicated that the suppression of the BAC by daidzin was due mainly to delay of stomach emptying. All 3 compounds suppressed voluntary alcohol consumption in alcohol-preferring rats. The decrease in alcohol consumption was accompanied by an increase in water intake, so that the total volume of liquid consumed daily remained unchanged. Daily food consumption and body weight gain were not affected. Alcohol preference returned to baseline levels after the isoflavonoids were discontinued. We postulate that the suppression of alcohol reinforcement produced by these compounds is mediated centrally in the brain reward pathway. PMID- 9848527 TI - Generalizability of the added benefits of guided tissue regeneration in the treatment of deep intrabony defects. Evaluation in a multi-center randomized controlled clinical trial. AB - BACKGROUND: Several studies have shown that GTR therapy of intrabony defects results in significantly better outcomes than access flap alone. Most of the available data, however, have been produced in highly controlled research environments by a small group of investigators. Generalizability of results to different clinicians and different subject populations has not been evaluated so far. METHODS: This parallel group study involved 143 patients recruited in a practice-based research network of 11 offices in 7 countries. It was designed to evaluate: 1) the applicability of the documented added benefits of GTR in the treatment of intrabony defects to different populations, and 2) the generalizability of the expected results to different clinicians. GTR was compared to access flap alone. Defects, one in each patient, were accessed with a previously described papilla preservation flap in both the test and control group. In addition, GTR sites received application of a bioabsorbable poly-D,L lactide-co-glycolide membrane. A stringent plaque control regimen was enforced in all patients during the 1-year observation period. Outcomes included gains in clinical attachment (CAL) and reductions in probing depth. RESULTS: Observed gains in CAL were 2.18 +/- 1.46 mm for access flap and 3.04 +/- 1.64 mm for the GTR-treated group. The treatment-associated difference was statistically significant (P = 0.03) after correcting for both center effect and defect anatomy. Among the various centers, a 1.73 mm difference in CAL gain was observed. This is a clinically relevant amount, which underlines the significance of center variability in the outcome of periodontal surgical procedures. A frequency distribution analysis of the obtained CAL gains indicated that GTR treatment of deep intrabony defects decreased, with respect to the access flap control, the probability of obtaining only a modest attachment gain at 1 year. Conversely, CAL gains of 4 mm or more were observed in more than 40% of GTR treated defects and in less than 20% of the controls (P < 0.0001). CONCLUSIONS: These data indicate that GTR therapy of deep intrabony defects performed by different clinicians on various patient populations resulted in both greater amounts and improved predictability of CAL gains than access flap alone. PMID- 9848528 TI - Early bacterial accumulation on guided tissue regeneration membrane materials. An in vivo study. AB - The aim of the study was to compare the in vivo early bacterial plaque colonization of 3 different guided tissue regeneration (GTR) membrane materials using a morphological (scanning electron microscope) method. Rectangular-shaped strips were cut from 3 periodontal membranes (expanded polytetrafluoroethylene, polyglactin 910, and polylactic acid) and glued to the buccal aspect of removable acrylic devices, which were applied to the molar-premolar region of the upper quadrants in 8 dental students. Each device held 3 strips: one ePTFE, one polyglactin 910, and one polylactic acid. The surface roughness of each membrane material was measured by means of a laser profilometer. During a 24-hour period, the students had to refrain from any oral hygiene procedures and did not use chlorhexidine mouthrinses. In each subject, one device was removed after 4 hours and the other after 24 hours. After removal, the devices were placed in a 2.5% gluteraldehyde solution to fix the membranes, which were then processed for SEM analysis. Fifty-four microscopic fields (at 200x magnification) were randomly selected and analyzed in each strip. Magnification was increased to determine the presence of bacterial morphotypes. The presence or absence of bacteria was assessed in a binomial fashion. In such a system, the field was bacteria-positive when bacteria constituted the deposits covering the surface of the membrane. The microscopic field was considered bacteria-negative when no bacteria were present. Bacteria-positive fields showing rods and filaments as prevalent bacterial morphotypes were recorded as rod-positive fields. A different pattern of plaque accumulation was demonstrated on different membrane materials. The 4-hour results indicated a statistically significant difference (P = 0.008, ANOVA) in the proportion of bacteria-positive fields among the 3 membranes; a greater amount of bacteria was demonstrated on the ePTFE membrane compared to the other 2 membranes. At 24 hours, the difference in the proportion of bacteria-positive fields was statistically significant (P = 0.002, ANOVA); a lesser amount of bacterial plaque was present on the polylactic acid membrane compared to the ePTFE and polyglactin 910 membranes. No difference in the proportion of rod/bacteria-positive fields was demonstrated among the 3 membranes at either 4 or 24 hours. It was concluded that quantitative differences in early plaque accumulation on various membranes seem to be related to the textural and structural characteristics of the surface, which is not adequately represented by the surface Ra value measured with a profilometric instrument. PMID- 9848529 TI - The use of autogenous periosteal grafts as barriers for the treatment of intrabony defects in humans. AB - The purpose of this study was to evaluate the clinical effectiveness of a connective tissue graft including periosteum used as a barrier for guided periodontal tissue regeneration in interproximal bony defects. An open flap debridement of a comparable interproximal bony defect in the same patient was used as a control. This study was performed at 2 different clinical centers. Six paired defects were treated at one center, and 16 paired defects at the other. Reentry surgeries were performed at 6 months. Preoperative comparisons of control and experimental sites with respect to clinical parameters and osseous measurements were similar. Post-surgical experimental sites produced more gain in clinical attachment (1.25 mm on buccal and 1.25 mm on lingual sites at center A and 1.26 mm on buccal and 1.18 mm on lingual sites at center B) and osseous defect fill (1.84 mm on buccal and 2.00 mm on lingual sites at center A and 1.66 mm on buccal and 2.04 mm on lingual sites at center B) when compared to control sites. The results of this trial indicate that clinical resolution of interproximal periodontal defects can be obtained with periosteal grafts used as barriers. PMID- 9848530 TI - Periodontal surgery of vertical bony defects with or without synthetic bioabsorbable barriers. 12-month results. AB - The aim of the present study was to clinically and radiographically compare guided tissue regeneration (GTR) therapy with bioabsorbable polyglactin 910 barriers and conventional periodontal surgery in intrabony defects. In 26 patients with advanced periodontitis, 29 teeth exhibiting interproximal intrabony defects were treated; 15 by conventional periodontal surgery (control) and 14 by GTR (test). Before and 12 months after surgery, clinical parameters were assessed and standardized radiographs were taken. On the radiographs the distances from the cemento-enamel junction (CEJ) to the alveolar crest (AC), and the CEJ to the most apical extension of the bony defect (BD) were measured using a computer assisted analyzing device (LMSRT). Twelve months after surgery, 24 patients with 27 lesions were available for examination. For both methods statistically significant (P < 0.001) probing depth (PD) reduction (mean +/- standard deviation) of -4.49 +/- 1.94 mm (n = 13, test) and -3.22 +/- 1.48 mm (n = 14, control), as well as clinical attachment gain (CAL-V) of 3.41 +/- 1.59 mm (test) and 2.07 +/- 1.10 mm (control), was observed. Radiographic changes of the distance CEJ to AC of -0.95 +/- 1.72 mm (n = 9, test), and -0.98 +/- 1.53 mm (n = 11, control) were not significant. A significant bony fill (distance CEJ-BD) of 1.05 +/- 1.22 mm was observed for the test group (P < 0.01); the 0.68 +/- 2.04 mm bony gain for the control group was not statistically significant. The PD reduction (P < 0.05) and attachment gain (P < 0.01) in the test group was statistically significantly more favorable than in the control group. Twelve months after surgery, statistically more favorable PD reduction and attachment gain was observed using polyglactin 910 barriers than compared to conventional flap surgery. Hence, the use of bioabsorbable barriers for therapy of intrabony defects may be recommended. PMID- 9848532 TI - Effects of bioabsorbable and non-resorbable barrier membranes on bone augmentation in rabbit calvaria. AB - The aim of this study was to compare the effects of bioabsorbable and non resorbable membranes on experimental guided bone augmentation in 8 Japanese white rabbits. A cutaneous flap was demarcated and raised from the forehead of each animal, the periosteum was lifted, and the calvarial bone on both sides of the midline was exposed. A titanium screw was inserted into the bone on each side of the midline and one screw was covered with a bioabsorbable (polylactic acid) membrane and the other with a non-resorbable (expanded polytetrafluoroethylene) membrane. The implanted screws and membranes were then covered with the periosteum and cutaneous flap. After healing for 6 months, the animals were euthanized and the experimental area was prepared for histological investigation. New bone had formed under both membranes with no sign of infection or membrane exposure. The amount of newly generated bone (89.0 +/- 17.3% versus 54.7 +/- 14.0%, P <0.05) and the percentage of newly generated bone height (81.5 +/- 6.3% versus 58.9 +/- 7.8%, P <0.05) in the space beneath the non-resorbable membrane was greater than that beneath the bioabsorbable membrane. However, there were no statistically significant differences between the bioabsorbable and non resorbable membranes with respect to the percentage areas of mineralized bone (52.3 +/- 11.3% versus 47.1 +/- 6.7%, P = 0.8658) and bone marrow (47.7 +/- 11.3% versus 52.9 +/- 6.7%, P = 0.4838) and bone contact with the screw (88.3 +/- 6.9% versus 89.2 +/- 7.3%, P = 0.9999). In conclusion, at least within the limitations of this rabbit model, we suggest that non-resorbable membranes with sufficient stiffness should be used to obtain greater bone volume and height instead of bioabsorbable membranes for the GBR procedure, and that this will facilitate predictable bone augmentation in spaces beyond the bone surface. Therefore, the bioabsorbable membrane could not replace the non-resorbable membrane used in this model. PMID- 9848531 TI - Guided tissue regeneration with bioabsorbable and expanded polytetrafluoroethylene barrier membranes in the treatment of naturally occurring periodontal defects in dogs. AB - The purpose of the present study was to evaluate the healing events and compare the effects of guided tissue regeneration (GTR) using 3 different membranes: a weakly cross-linked bioabsorbable bovine collagen membrane; a strongly cross linked bioabsorbable bovine collagen membrane; and a non-resorbable ePTFE membrane. Twenty adult female beagle dogs with naturally occurring periodontitis were subjected to periodontal probing and scaling. In both right and left mandibular quadrants, mucoperiosteal flaps were raised, and after debridement, the roots of experimental premolar teeth received apical reference notches. The following treatments were randomly assigned: 1) gingival flap curettage (GFC) + ePTFE membrane, control membrane; 2) GFC + slightly cross-linked collagen membrane, 1st test membrane; and 3) GFC + strongly cross-linked collagen membrane, 2nd test membrane. The flaps were sutured in such a manner that the membranes were completely covered. All dogs received a soft diet for a 2-week period and an oral hygiene program until time of sacrifice. The animals were randomly scheduled for sacrifice at 2, 4, 12, and 24 weeks. The ePTFE membranes still in place were removed at 6 weeks. The jaws were dissected and specimens prepared for descriptive histology and histomorphometry. The early resorption of the 1st test membrane was achieved at 4 weeks, and the 2nd test membrane at 12 weeks, both with normal inflammatory reaction. Measurements of epithelium, connective tissue attachment, new bone, and neocementum were compared within an animal (paired t test). Analyses were performed on data at 4, 12, and 24 weeks post-healing; little differences were found between these periods. Limited connective tissue repair was favored by the placement of all the membranes (about 20%), with no statistically significant difference. These findings indicate that bioabsorbable collagen membranes with different cross-linking and ePTFE barriers promote similar new attachment in GTR procedures on naturally occurring periodontal defects in dogs. PMID- 9848533 TI - Evaluation of a diphenylphosphorylazide-crosslinked collagen membrane for guided bone regeneration in mandibular defects in rats. AB - In the present study, the potential of a diphenylphosphorylazide-crosslinked type I bovine collagen membrane was evaluated in the healing of mandibular bone defects applying the biological concept of guided bone regeneration. The experiment was carried out on 25 Wistar rats. After exposing the mandibular ramus bilaterally, 5 mm diameter full-thickness circular bone defects were surgically created. While the defect on one side was covered by the membrane (experimental), the defect on the other side was left uncovered (control) before closure of the overlying soft tissues. The rats were sacrificed in groups of 5 after 7, 15, 30, 90, and 180 days of healing. Although at early stages of healing similar amounts of bone formation were observed in the experimental and control defects, after 1 month of healing, most of the experimental defects were completely closed with new bone, while in the control defects, only limited amounts of new bone were observed at the rims and in the lingual aspect of the lesions. In the 90- and 180 day animals, all experimental defects were completely closed, while in the control defects, no statistically significant increase in bone regeneration was observed. The increase in percentage of bone regeneration in the experimental defects was statistically significant between the 15-day specimens as compared with the 7-day specimens (P < 0.01) and likewise between 30-day and 15-day specimens (P < 0.001). It can be concluded that a DPPA-crosslinked collagen membrane yields biocompatibility, ad hoc mechanical hindrance, and handling characteristics suitable for guided bone regeneration applications in this experimental model. PMID- 9848535 TI - Isolation of human osteoblast-like cells and in vitro amplification for tissue engineering. AB - As the field of dental implants continues to grow at a rapid rate so does our quest to find new techniques to enhance bone grafting. Tissue engineering is an exciting new technique in bone grafting. Therefore, the purposes of this study were to develop a simple, reproducible method to isolate human osteoblast-like cells (HOBs) and to evaluate in vitro cell proliferation within 2 different 3 dimensional (3-D) constructs targeted for tissue engineering applications. Ultimately, HOBs that have been amplified within 3-D constructs may be employed for bone regeneration techniques, such as onlay and sinus grafting prior to implant placement. Our cell isolation protocol employed human fetal calvaria tissue sequentially digested with trypsin and collagenase. The HOB cells from only the third and fourth digests were obtained, cultured and evaluated within the constructs. An osteoblast-like phenotype was in part verified for these HOB cells by demonstrating a significantly higher alkaline phosphatase activity than for human gingival fibroblasts, and a comparable level to the osteoblast cell line MG-63. The HOB cells were cultured within either poly (D,L-lactide) (PLA) or a fused fiber ceramic and evaluated for the ability to support in vitro HOB amplification. HOB proliferation was validated by scanning electron microscopy, identifying cells throughout the 3-D constructs. Continuous cell viability was demonstrated for the duration of the 33-day evaluation period and the extent of cell amplification reached approximately 20 times the seeding density. The in vitro amplification results further indicate that tissue engineering strategies with either the PLA or fused fiber construct may be suitable for bone regeneration therapy for dental implants. PMID- 9848534 TI - Protein extracts of dentin affect proliferation and differentiation of osteoprogenitor cells in vitro. AB - Proteins associated with the mineral phase of dentin are considered to have the potential to alter cell function within the local environment, during development and regeneration of tooth/periodontal tissues. Cells that may be altered include osteoblasts, ameloblasts, periodontal ligament cells, odontoblasts, and cementoblasts. However, specific factors within dentin controlling cell activity have not been elucidated. To investigate further the role of dentin proteins in regulating cell behavior, MC3T3-E1 cells, a mouse osteoprogenitor cell line, were exposed to guanidine/EDTA extracts of dentin (G/E-D) prepared from bovine teeth. Cells, with or without G/E-D (2 to 50 microg/ml), were evaluated for proliferative activity and for mRNA expression of bone-associated genes. Results indicated that G/E-D suppressed cell proliferation and caused striking morphological changes, including the conversion of cuboidal cells into fibroblastic, spindle-shaped cells. Markers of osteoblast differentiation, osteocalcin and bone sialoprotein mRNA were decreased, while osteopontin mRNA was enhanced in cells exposed to G/E-D. Since transforming growth factor beta (TGFbeta1) has been reported to influence cells in a similar fashion, G/E-D were examined for the presence of and concentration of TGFbeta using slot blot analysis and enzyme immunoassay (ELISA), respectively. These analyses demonstrated that G/E-D contained 6.6 ng/mg of TGFbeta1. Next, cells were exposed to G/E-D in conjunction with anti-TGFbeta1,2,3 antibody. When cells were exposed to antibody, G/E-D-mediated changes in morphology and gene expression were blocked. These results suggest that TGFbeta1 and perhaps other factors in dentin can regulate cell behavior and, therefore, can influence development, remodeling, and regeneration of mineralized tissues. PMID- 9848536 TI - Effects of Porphyromonas gingivalis 2561 extracts on osteogenic and osteoclastic cell function in co-culture. AB - This study was undertaken to determine the direct effects of extracts derived from Porphyromonas gingivalis on bone formation and mineral resorption in an osteogenic/osteoclastic cell in vitro co-culture model. Osteogenic bone marrow derived stromal cells were isolated from 18-day old embryonic chickens, while osteoclastic cells were isolated from laying white Leghorn hens on calcium deficient diets. Osteoclastic cells (5 x 10(5)) were seeded onto mineral thin films and suspended above osteogenic cells (1 x 10(4)) already plated on the bottoms of tissue culture plate wells. Sonicated P. gingivalis 2561 extracts were prepared from whole bacterial cells and added in varying proportions (0 to 2 microg/ml) to the co-culture growth medium. These co-cultures, and appropriate mono-culture controls, were incubated for a further 4 days. Parameters of bone forming cell activity including alkaline phosphatase activity, calcium and inorganic phosphate accumulation were performed on the osteogenic cells. Mineral substrate resorption by osteoclastic cells was assessed morphometrically. In their respective mono-cultures, the addition of P. gingivalis sonicate to the culture medium had no effect on osteoclastic mineral resorption, but significantly inhibited osteogenesis (up to 45%; P <0.05). In co-cultures, however, the sonicate induced significant increases in mineral resorption (up to 70%; P <0.05), whereas bone forming cell activity was still inhibited, although to a significantly lesser extent than in mono-cultures (up to 25%; P <0.05). These results suggest that P. gingivalis sonicate induced up-regulation of mineral resorption may be mediated via osteogenic cells. PMID- 9848537 TI - Subpedicle connective tissue graft versus guided tissue regeneration with bioabsorbable membrane in the treatment of human gingival recession defects. AB - The purpose of the present clinical study was to evaluate the effect of guided tissue regeneration (GTR) in comparison to subpedicle connective tissue graft (SCTG) in the treatment of gingival recession defects. A total of 12 patients, each contributing a pair of Miller's Class I or II buccal gingival recessions, was treated. According to a randomization list, one defect in each patient received a polyglycolide/lactide bioabsorbable membrane, while the paired defect received a SCTG. Treatment effect was evaluated 6 months postsurgery. Clinical recordings included full-mouth and defect-specific oral hygiene standards and gingival health, recession depth (RD), recession width (RW), probing depth (PD), clinical attachment level (CAL), and keratinized tissue width (KT). Mean RD significantly decreased from 3.1 mm presurgery to 1.5 mm at 6 months postsurgery for the GTR group (48% root coverage), and from 3.0 mm to 0.5 mm for the SCTG group (81% root coverage). RD reduction and root coverage were significantly greater in SCTG group compared to GTR group. Mean CAL gain amounted to 1.7 mm for the GTR group, and 2.3 mm in the SCTG group. No significant differences in PD changes were observed within and between groups. KT increased significantly from presurgery for both treatment groups, however gingival augmentation was significantly greater in the SCTG group compared to GTR group. Results indicate that: 1) treatment of human gingival recession defects by means of both GTR and SCTG procedures results in clinically and statistically significant improvement of the soft tissue conditions of the defect; and 2) treatment outcome was significantly better following SCTG compared to GTR in terms of recession depth reduction, root coverage, and keratinized tissue increase. PMID- 9848538 TI - Change in temperature of subjacent bone during soft tissue laser ablation. AB - In tissues that closely approximate bone, sufficient heat may be transferred to the bone during laser surgery to cause damage and/or necrosis. To date, there have been few studies examining the temperatures elicited at the bone surface as a result of laser application to the overlying soft tissues. The purpose of this investigation was to determine, under in vitro conditions, temperature changes at the bone/soft tissue interface during laser ablation with CO2 and Nd:YAG lasers used with and without (w/wo) air/water coolant. Experimental specimens consisted of 5 mandibles from freshly sacrificed hogs; laser treatment sites were the buccal and lingual attached gingiva of the molars and the lingual keratinized mucosa of the incisor region. CO2 and Nd:YAG lasers were used w/wo coolant at power settings of 4 to 8 W and 5 to 9 W, respectively. Temperature changes were measured with a copper constant thermocouple contained within a 21 gauge hypodermic needle. In comparing the lasers at comparable energy densities w/wo coolant, temperature increases at the bone/soft tissue interface ranged from 8.0 to 11.1 degrees C with the Nd:YAG and 1.4 to 2.1 degrees C with the CO2. Similarly, in comparing the times required for the interface to return to baseline temperature following removal of the laser, values ranged from approximately 143 to 205 and approximately 119 to 139 seconds for the Nd:YAG and CO2, respectively. Results from this study suggest that, at energy densities equal or above those reported here, the increase in temperature at the bone surface as a result of periodontal soft tissue surgery with the Nd:YAG laser could be damaging, especially if the exposure is prolonged. PMID- 9848539 TI - Scanning electron microscopic study of the effects of Er:YAG laser on root cementum. AB - Use of Er:YAG laser has been proposed for the removal of microbial deposits and calculus present on teeth affected by periodontal disease. However, the influence of Er:YAG laser irradiation on root surfaces has not yet been fully investigated. The aim of the present study was to evaluate the effects of Er:YAG laser irradiation on root cementum by scanning electron microscopy (SEM). Specimens were obtained from extracted human periodontally-diseased teeth using a water cooled high-speed bur. An Er:YAG laser beam was then applied at various powers ranging from 25 to 100 mJ/ pulse/sec. The laser irradiation was performed under water irrigation, with the tip held perpendicular to the root surface in the contact mode. Following laser exposure, specimens were fixed, dehydrated, and dried at critical-point in liquid CO2. After mounting on SEM plates and sputter coating with gold, the cementum surface was examined by SEM. Observations of the root surface showed a relatively flat surface in control specimens. In Er:YAG exposed specimens, the laser beam created a circular, notched-edge, crater-like defect on the root. The bottom of the lesion showed an irregular and sharp pointed surface. Subsequently, the specimens were fractured with a sharp scalpel perpendicularly to the surface. SEM observations of these specimens showed a 15 microm layer of damaged tissue within the laser-irradiated cementum. The tissue presented an amorphous appearance and the Sharpey's and matrix fiber bundles were not clearly distinguishable. These observations indicate that cementum tissue could be damaged by Er:YAG laser irradiation. PMID- 9848540 TI - Effects of metronidazole plus amoxicillin in progressive untreated adult periodontitis: results of a single 1-week course after 2 and 4 months. AB - A randomized, double-blind, clinical study was done to assess the microbiological and clinical effects of metronidazole plus amoxicillin (M+A) as the only therapy in 46 patients with moderate to advanced progressive adult periodontitis. Patients were included in the study after at least 2 sites showed > or =2 mm clinical attachment loss. Bleeding on probing, probing depth, and clinical attachment level were measured using on automated probe. The percentage of surfaces with plaque was recorded at day 0, and at 2 and 4 months after therapy. No effort was made to change the oral hygiene habits of patients. Identification of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia was assessed utilizing DNA technology at day 0 and 2 months after therapy. Twenty-three patients received metronidazole 250 mg plus amoxicillin 500 mg, 3 times/day for a week and 23 a placebo. Two patients in the placebo group were dropped at 2 months because they had taken antibiotics for medical reasons. Statistical analyses of differences between groups was done using the Mann Whitney test, and the differences within each group were tested with ANOVA. There were no significant changes in surfaces with plaque in either group after therapy. The percentage of bleeding sites decreased significantly from baseline to 2 and 4 months in the M+A group (P = 0.001), and increased in the placebo group. Differences in bleeding on probing between groups were significant at 2 (P = 0.018), and 4 months (P = 0.005). The mean attachment level values at 2 and 4 months post-therapy improved significantly in the M+A group compared to the placebo group (P = 0.001). Treatment with M+A resulted in a significant mean reduction in probing depth at 2 and 4 months compared to baseline values (P = 0.001). The M+A group showed a significant reduction of sites with high levels of Pg (P = 0.001) at 2 months compared with baseline values, and there was a significant reduction of sites with Pg and Pi in the M+A group compared with the placebo group. The results showed that a combined M+A treatment as the only therapy changes the proportion of some subgingival microorganisms and allows a significant improvement in clinical conditions. PMID- 9848541 TI - The correlation of gingival crevicular fluid interleukin-8 levels and periodontal status in localized juvenile periodontitis. AB - Localized juvenile periodontitis (LJP) is an early-onset periodontal disease characterized by progressive bone loss involving the permanent first molar and incisor teeth. Approximately 70% to 75% of LJP patients have impaired neutrophil chemotaxis towards a number of chemoattractants including N-formyl-methionyl leucyl-phenyl-alanine, complement fragment C5a, leukotriene B4, and interleukin 8 (IL-8). The aim of the present study was to observe the role of IL-8 in the pathogenesis of LJP. Fourteen individuals who were systemically and periodontally healthy and 24 systemically healthy individuals diagnosed with LJP (based on the results of clinical periodontal assessments and radiographic examination) were recruited for this study. Gingival crevicular fluid (GCF) samples were obtained from anterior teeth in each subject before treatment. After evaluation of GCF amount from paper strips, enzyme-linked immunoabsorbent assay was employed to determine the amount of IL-8 in GCF. The amount and concentration of IL-8 measured was 894.5 +/- 435 pg, and 445.3 +/- 468 pg/microl for the experimental group and 747.3 +/- 543 pg and 684.7 +/- 548 pg/microl, for the control group. The correlation among the levels of cytokine and clinical parameters was assessed. It was observed that the concentration of IL-8 demonstrated a negative correlation with gingival index in the LJP group. In addition, no significant correlation was found among the total amount and concentration of IL-8, GCF volume, and clinical parameters in the control group. IL-8 is thought to enhance host defense mechanisms against gram-negative bacteria, thus providing protection against periodontal infections. Our data demonstrate that, when both the total amount and concentration of IL-8 are taken into consideration, no significant difference between LJP and healthy subjects is shown. This may indicate a less active IL-8 production compared with healthy subjects in spite of the dense Gram bacterial stimulation in LJP. PMID- 9848542 TI - Root coverage with a connective tissue with partial thickness double pedicle graft and an acellular dermal matrix graft: a clinical and histological evaluation of a case report. AB - Root coverage can be achieved by many techniques. In this case report a patient was treated with a connective tissue with partial thickness double pedicle graft, which resulted in complete root coverage. However, 6 days postsurgery the patient developed a bleeding problem from the palatal donor area. For this reason, a unique approach was used to treat the next area needing root coverage. The procedure combined an acellular dermal matrix and a coronally positioned pedicle. The procedure was used to treat 3 defects. Complete root coverage was obtained on 2 of the 3 defects. The third was covered to within 1 mm of the cemento-enamel junction (CEJ). Histology showed similar results with a connective tissue graft and an acellular dermal matrix. Verhoeff's staining demonstrated that the acellular dermal matrix was incorporated into the gingival tissue. This case report demonstrated that acceptable results can be obtained with the connective tissue with partial thickness double pedicle graft and the acellular dermal matrix combined with a coronally positioned pedicle, both clinically and histologically. PMID- 9848543 TI - Re: Comparison of porous bone mineral and biologically active glass in critical sized defects (1997;68:1043-1053) PMID- 9848545 TI - The journal club and its role in hand surgery education. AB - Journal clubs are a time-honored method of teaching literature appraisal skills in many residency programs. A questionnaire was completed by 57 hand surgery fellowship directors and 91 hand fellows to define the role of the journal club in hand fellowship training. We sought to identify definitive characteristics of hand surgery journal clubs and assess their strengths and weaknesses. One hundred forty-eight of 164 (90%) directors and fellows responded, yielding information on 57 of 58 accredited hand fellowships. Forty-nine of 57 (86%) responding fellowships have a journal club. The majority of clubs meet monthly for 1 to 2 hours, usually within the hospital. The primary purpose is to familiarize both fellows and attendings with the current literature. Most often, fellows choose the articles, which are most commonly original research and review articles. Usually, faculty presides, and residents and fellows present. Increased faculty participation was the most frequently suggested improvement. The great majority of those surveyed felt that their journal club was successful and was an important part of the fellowship training. We conclude that journal clubs have a high perceived value by participants, and recommend the journal club to all hand surgery communities, with or without fellowship involvement. PMID- 9848544 TI - Re: Periodontal management of gingival overgrowth in the heart transplant patient: a case report (1997;68:1140-1146) PMID- 9848546 TI - An anatomic study of the ligamentous structure of the triangular fibrocartilage complex. AB - An anatomic study of the ligamentous structures of the triangular fibrocartilage complex and their attachments on the ulnar styloid was performed using 27 embalmed cadaver wrists. The dorsal and palmar distal radioulnar ligaments of the triangular fibrocartilage complex in each specimen contained a superficial and a deep portion. The deep portion of both ligaments inserted on the fovea of the ulna. The superficial portion of both ligaments surrounded the articular disc uniting at the ulnar-most portion of the articular disc. The tissue that is between the ulnar aspect of the superficial ligament (and integrated on its periphery) and the ulnar capsule is defined as the meniscus homologue. Anatomic variations in the meniscus homologue and the prestyloid recess (the cavity adjacent to the ulnar styloid) were seen in 1 of 3 ways; the narrow opening type in 74% of specimens, the wide opening type in 11%, and the no opening type in 15%. The ulnotriquetral ligament inserted on the palmar-radial aspect of the base of the ulnar styloid and the ulnolunate ligament inserted on the palmar border of the articular disc. PMID- 9848547 TI - The triangular fibrocartilage complex: an important component of the pulley for the ulnar wrist extensor. AB - The extensor carpi ulnaris (ECU) tendon is the only wrist motor tendon that broadly connects with the triangular fibrocartilage complex (TFCC) of the wrist. The goal of this study was to determine the biomechanical effect of the TFCC on the function of the ECU. The effect of avulsion of the TFCC on the changes in mechanics of the ulnar wrist extensor tendon was investigated in 8 fresh-frozen cadaver forearms. Excursion of the ECU tendon was continuously recorded over the functional range of wrist extension and ulnar deviation in intact wrists, wrists with ulnar styloid fractures, wrists with TFCC release from the distal ulna, and after excising the distal ECU tendon sheath. The ECU tendon demonstrated a 30% increase in excursion during wrist extension after release of the TFCC from its attachment on the distal ulna. During 60 degrees of wrist extension, excursion of the ECU tendon was 4.8+/-1.9 mm in the intact wrists and 6.3+/-2.0 mm after TFCC release. This change in excursion represented 1.4 mm of bowstringing for the ECU tendon during 60 degrees of wrist extension. Further incision of the distal part of the extensor sheath produced only 6% increase in excursion of the ECU. Results of this study suggest that the TFCC is an important component of the pulley for the ulnar wrist extensor. These findings imply that disturbance of the wrist extensor after TFCC injury may potentially contribute to abnormal loading and force transmission through the ulnar wrist and the TFCC, and support the growing consensus that integrity of the TFCC should be restored in the presence of TFCC injuries. PMID- 9848548 TI - Ulnar nerve strains at the elbow. AB - Twenty unembalmed intact cadavers had strain measurements taken of the ulnar nerve at 5 positions around the elbow and at varying degrees of elbow flexion. A microstrain gauge was implanted directly into the nerve through a superficial incision. Maximum strains in the ulnar nerve occurred at maximum elbow flexion and directly behind the medial epicondyle; meaningfully less strain was seen at lesser degrees of flexion and positions distal and proximal to the epicondyle. A wide range of maximal strains (0% to 14%) was found. Twenty-five percent of the ulnar nerves showed strains of greater than 10% with maximum elbow flexion behind the medial epicondyle. Cubital tunnel syndrome may be at least in part a traction neuropathy. PMID- 9848549 TI - Moment arms of the digital flexor tendons at the wrist: role of differential loading in stability of carpal tunnel tendons. AB - When a flexor digitorum superficialis tendon crossing a flexed or extended wrist has a load applied to it in excess of that applied to adjacent tendons, that tendon may translate across the carpal tunnel. In 6 cadaver specimens, each of the 9 carpal tunnel tendons was loaded with a baseline tension of 85 g and the moment arms of the flexor pollicis longus and the 4 flexor digitorum superficialis tendons were determined. Applying a higher 540-g load to individual flexor digitorum superficialis tendons and the flexor pollicis longus while loading the remaining tendons with the baseline 85-g tension significantly changed the moment arms from those measured under baseline load. The results demonstrated that tendons with applied differential loads in the carpal tunnel shift their positions, as revealed by their changing moment arms. PMID- 9848550 TI - Effects of finger posture on carpal tunnel pressure during wrist motion. AB - Persistent elevations in carpal tunnel pressure may aggravate carpal tunnel syndrome. This study examined the effects of finger posture on carpal tunnel pressure during wrist motion. Carpal tunnel hydrostatic pressure was measured using a saline-filled catheter inserted into the nondominant wrists of 14 healthy individuals. Range of motion tasks of wrist flexion-extension and radioulnar deviation were repeated with metacarpophalangeal (MCP) joint angles of 0 degrees, 45 degrees, and 90 degrees flexion. Pressures were significantly greater with the fingers straight (MCP = 0 degrees) than when the MCP joints were flexed to 45 degrees for all radioulnar deviation angles and from 10 degrees of wrist flexion to all angles of wrist extension tested. Pressures were also significantly higher with MCP joints at 0 degrees than at 90 degrees for wrist extension angles from 10 degrees to 40 degrees. Pressures increased to over 30 mm Hg (4.0 kPa) in some wrist extension and ulnar and radially deviated postures. Finger and wrist postures should be considered when designing splints or evaluating tasks for patients with carpal tunnel syndrome. PMID- 9848551 TI - Persistent or recurrent carpal tunnel syndrome following prior endoscopic carpal tunnel release. AB - A retrospective review of patients with carpal tunnel syndrome (CTS) identified 20 patients (26 wrists) who had persistent or recurrent CTS after having undergone endoscopic carpal tunnel release (ECTR). Seven wrists never had relief of the original CTS symptoms; for the remaining 19 wrists, the average time to recurrence of CTS following ECTR was 4.8 months. All cases recurred within 12 months from the ECTR. All but 1 patient reported nocturnal pain. All patients experienced morning numbness and stiffness. All wrists had positive Phalen's test results. Once diagnosed with persistent or recurrent CTS, each of the wrists underwent open carpal tunnel release (OCTR). Twenty-two wrists were available for follow-up, with all patients reporting greater satisfaction and relief after the OCTR compared with the ECTR. It is important to recognize that CTS may persist or recur after ECTR. These patients should be treated with OCTR to provide symptomatic relief and prevent the sequelae of ongoing CTS. PMID- 9848552 TI - Histology of the transverse carpal ligament and flexor tenosynovium in idiopathic carpal tunnel syndrome. AB - Histology of the transverse carpal ligament and flexor tenosynovium was investigated in 166 wrists from 130 patients with idiopathic carpal tunnel syndrome. Nine wrists from 9 controls were used for comparison. In the patients, 73.5% of the ligament and 56.0% of the tenosynovium showed normal histology. In the remaining specimens, the ligament showed mucoid change (17.5%), amyloid deposits (7.8%), inflammation (3.0%), and chondrometaplasia (2.4%), and the tenosynovium showed fibrosis (25.3%), edema (7.2%), inflammation (10.2%), amyloid deposits (8.4%), and vascular hypertrophy (4.8%). The majority of these changes were mild in degree and the pathophysiology did not appear to be meaningful. We conclude that in idiopathic carpal tunnel syndrome, the ligament and tenosynovium often show normal histology and there are no typical or consistent changes with which idiopathic carpal tunnel syndrome can be associated. PMID- 9848553 TI - The effect of denervated muscle and Schwann cells on axon collateral sprouting. AB - The effects of denervated muscle and Schwann cells on collateral sprouting from peripheral nerve were studied in the peroneal and tibial nerves of 48 Sprague Dawley rats. Three groups were prepared. In group MSW (muscle-Schwann cell window), the peroneal nerves were transected 3 mm below the sciatic bifurcation. The proximal stumps were sealed in a blocked tube to prevent regeneration and the distal stumps were implanted into denervated muscle cells that were wrapped around the ipsilateral tibial nerve, which had a window of perineurium resected. Schwann cells from the ipsilateral sural nerve were implanted into the muscle. Group MS (muscle-Schwann cell) was similar to group MSW, except that the tibial nerve perineurium was kept intact. In group MW (muscle-window), the muscle was prepared without Schwann cells and the tibial nerve perineurium was windowed. S 100 immunostain was used to identify the Schwann cells surviving 1 week after transplantation. After 16 weeks of regeneration, horseradish peroxidase tracer was used to label motor neurons and sensory neurons reinnervating the peroneal nerve. Myelinated axons of the reinnervated peroneal nerves were quantified with the Bioquant OS/2 computer system (R&M Biometrics, Nashville, TN). A mean of 169 motor neurons in group MSW, 64 in group MW, and 26 in group MS reinnervated the peroneal nerve. In the dorsal root ganglion, the mean number of labeled sensory neurons was 1,283 in group MSW, 947 in group MS, and 615 in group MW. The mean number of myelinated axons in the reinnervated peroneal nerve was 1,659 in group MSW, 359 in group MS, and 348 in group MW. Reinnervated anterolateral compartment muscles in group MSW were significantly heavier than those in group MS or MW. This study demonstrates that the transplantation of denervated muscle and Schwann cells promotes motor and sensory nerve collateral sprouting through a perineurial window. PMID- 9848554 TI - Intraoperative measurement of choline acetyltransferase activity to evaluate the functional status of donor nerve during reinnervated free muscle transfer: a preliminary report. AB - Intraoperative choline acetyltransferase activity measurements were used to evaluate the functional status of donor nerves during reinnervated free muscle transfer. This technique was applied to 3 cases. One was a lower-type brachial plexus injury in combination with radial nerve injury; the other 2 were Volkmann's ischemic contractures. Gracilis muscle was transferred for reconstruction of wrist extension in all cases. The donor nerves included 1 anterior interosseous nerve and 2 posterior interosseous nerves. A fascicle with choline acetyltransferase activity above 2,000 cpm was considered to be reliable as a motor fascicle. Reinnervation of transferred muscle was confirmed by electromyographic examination within the first 3.3 months (range, 2.5 to 4 months) after surgery and all muscles obtained useful recovery. This technique can directly and quantitatively verify the functional quality of the donor motor fascicle when the quality of the donor nerve is in doubt. PMID- 9848555 TI - The motion path of the digits. AB - This study investigates whether the path taken by the fingertips of the human hand during unrestricted flexion and extension follows a precise mathematical pattern: an equiangular spiral. Eight normal subjects participated in the study. Subjects performed numerous flexion and extension trials at a random speed. Motion was recorded by a 6-camera, 3-dimensional motion analysis system with 24 retroreflective markers affixed to the dominant hand at predetermined locations. Four hundred eighty flexion-extension arcs were analyzed. We used the coefficient of multiple determination to compare the flexion and extension motion arc of each finger to an equiangular spiral curve derived mathematically. Our results indicate that the path of the hand during flexion and extension closely follows the path of an equiangular spiral with the coefficient of multiple determination values consistently above 0.95. PMID- 9848556 TI - A randomized biomechanical study of zone II human flexor tendon repairs analyzed in a linear model. AB - Komanduri et al showed that dorsal tendon repairs using Kessler and Bunnell techniques were stronger than the standard volar repair (J Hand Surg 1996;21 A:605-611). They concluded that when testing in the anatomic curvilinear mode, the differences in strength were due to tension banding. Soejima et al challenged that concept by stating that the difference in strength was in the biomechanics of the dorsal tendon itself (J Hand Surg 1995;20A:801-807). We set out to confirm Soejima et al's theory by using more core suture techniques. We compared the tensile strength at 2-mm gap and the ultimate tensile strength of Kessler, Strickland, Robertson, and modified Becker sutures. Ten repairs of each suture type were placed either dorsally or volarly in matched human cadaver flexor tendons. There was no statistical difference between volar and dorsal placement for either maximum tensile force or force at 2-mm gap. Our study does not confirm Soejima et al's in any of the four suturing techniques. PMID- 9848557 TI - A randomized biomechanical study of zone II human flexor tendon repairs analyzed in an in vitro model. AB - To study the effect of dorsally placed flexor tendon repairs on human cadaver tendons, we used the Kessler, Strickland, Robertson, and modified Becker techniques. The strengths of the repairs were tested in a previously described human cadaveric curvilinear model. There were statistically significant increases in dorsal versus volar grasping strength with the Kessler repair (38 N vs 33 N) and the Robertson repair (51 N vs 43 N). There were no differences with the locking Strickland and modified Becker repairs. One may infer that the locking techniques are intrinsically tighter and may not be as susceptible to the different forces between the tensile and compressive surfaces of a dynamic flexor tendon. PMID- 9848558 TI - Molecular studies in flexor tendon wound healing: the role of basic fibroblast growth factor gene expression. AB - Basic fibroblast growth factor (bFGF) is a cytokine that plays a fundamental role in angiogenesis. This study examines bFGF messenger RNA (mRNA) expression in a rabbit flexor tendon wound healing model. Thirty-four New Zealand white rabbit forepaws underwent transection and repair of the middle digit flexor digitorum profundus tendon in zone II. Tendons were harvested at increasing time intervals and analyzed by in situ hybridization and immunohistochemistry. Few tenocytes and tendon sheath cells expressed bFGF mRNA in unwounded tendons. In contrast, tendons subjected to transection and repair exhibited an increased signal for bFGF mRNA in both resident tenocytes concentrated along the epitenon and infiltrating fibroblasts and inflammatory cells from the tendon sheath. These data demonstrate that (1) normal tenocytes and tendon sheath cells are capable of bFGF production, (2) bFGF mRNA is upregulated in the tendon wound environment, and (3) the upregulation of this angiogenic cytokine occurs in tenocytes as well as in tendon sheath fibroblasts and inflammatory cells. PMID- 9848559 TI - Treatment of interphalangeal hyperflexion and metacarpophalangeal hyperextension of the thumb in combined low median-ulnar nerve palsy. AB - A method of treatment of interphalangeal hyperflexion and metacarpophalangeal hyperextension of the thumb in combined low median-ulnar nerve palsy is described. The flexor pollicis longus tendon is divided longitudinally into 2 slips. One tendon strip is cut distally, extracted from the fibro-osseous canal, passed superficial to the tendon sheath, and attached to its previous insertion. An extensor indicis proprius opposition transfer was simultaneously performed. Eleven patients were monitored for 1 to 4 years after surgery. All results were positive, and there were no recurrences of either Froment's or Jeanne's sign. PMID- 9848560 TI - Surgical treatment of arthrogryposis of the elbow. AB - The purpose of this study was to analyze our results of surgical treatment of arthrogryposis of the elbow and to compare our tendon transfer results using range of motion (ROM) criteria versus functional use criteria. Eighteen tendon transfers for elbow flexion in 14 children with arthrogryposis with an average follow-up period of 4 years (range, 1-14 years) and 6 elbow capsulotomies with triceps lengthening in 6 children with arthrogryposis with an average follow-up period of 5 years (range, 2-9 years) were evaluated. Each child was assessed by a questionnaire regarding functional use of the upper extremity, physical examination of ROM and strength, and a videotaped activities of daily living evaluation. Tendon transfer results were classified and compared using 2 methods of evaluation: postoperative strength and ROM and effective functional use of the tendon transfer to perform activities of daily living. The 6 elbow capsulotomies improved from an average preoperative arc of 17 degrees of motion (average extension, -2 degrees; average flexion, 19 degrees) to an average final follow-up arc of 67 degrees (average extension, -25 degrees; average flexion, 92 degrees). The 18 tendon transfers evaluated by strength and ROM criteria showed 9 triceps to biceps transfers in 9 arms (7 good, 1 fair, and 1 poor), 5 pectoralis to biceps transfers in 4 arms (1 good, 3 fair, and 1 poor), and 4 latissimus dorsi to biceps transfers in 3 arms (2 good and 2 fair). Evaluation by functional use criteria gave the same result in 13 transfers and downgraded the result in 5; the downgraded results were due to resultant flexion contracture or limited functional use because the transfer was in the nondominant arm. Based on this review, optimal surgical candidates for tendon transfer are children older than 4 years, who have full passive ROM of the elbow in the dominant arm, and at least grade 4 strength of the muscle to be transferred. PMID- 9848561 TI - The radiographic analysis of web height. AB - Web creep is a common complication following surgical release of syndactyly. Currently, normal web height has not been objectively determined, which prevents accurate analysis of the degree of web creep after surgery. The purpose of this study was to design a technique to reproducibly measure web height and to define the standard web height in a control population. Four hundred thirty-seven standard left posteroanterior hand x-rays of children without any upper extremity pathology were evaluated. Web height was measured as a relative ratio to digital length using standard landmarks. All measurements were tabulated according to gender, age, and web space. Statistical analysis was performed to determine the reliability of this technique and to delineate any differences between web space, gender, and age. There was no significant intraobserver or interobserver difference and web height was similar in males and females. There was a significant difference between web height with respect to web location and age. Normal ranges of web height with regard to age and web position were determined and plotted as graphs. Using this measurement method, web creep can be evaluated and comparison studies of different surgical techniques performed. PMID- 9848562 TI - The significance of the three volar spaces in forearm compartment syndrome: a clinical and cadaveric correlation. AB - The goal of this study was to assess the importance of the 3 volar spaces in forearm compartment syndrome in a prospective manner. A cadaveric model was developed to correlate with our clinical experience. All but 1 of 21 volar compartments (superficial volar, deep volar, and pronator quadratus spaces) in 7 patients in our clinical series decompressed adequately after release of the superficial volar fascia. One patient needed further release of the pronator quadratus compartment; he had suffered a combined crush and vascular injury. All volar compartments in our cadaveric model decompressed with superficial fasciotomy. Prefasciotomy and postfasciotomy pressures should be obtained from all 3 compartments of the volar forearm. Superficial fasciotomy usually adequately decompresses the entire volar forearm; however, in the event that deep compartment pressures remain high after superficial fasciotomy, release of the affected space is indicated. PMID- 9848563 TI - The nonarticulating portion of the radial head: anatomic and clinical correlations for internal fixation. AB - The proximal radioulnar joint was dissected in 24 cadaveric elbows to localize the area of the radial head that did not articulate with the lesser sigmoid notch of the ulna. The nonarticulating portion of the radial head was posterolateral in the anatomic position of full supination. Gross observations of the nonarticulating portion of the radial head revealed a thinner band of yellowish cartilage relative to a wider, white, glistening cartilage of the articular portion of the radial head. The nonarticular portion of the radial head did not contain the angled peak, which is most prominent in the middle of the articular portion. The average arc of the nonarticulating radial head was 113 degrees (range, 106 degrees to 120 degrees; standard deviation, 4 degrees). This nonarticulating portion of the radial head (or safe zone for prominent fixation) consistently encompassed a 90 degrees angle localized by palpation of the radial styloid and Lister's tubercle. Using these palpable distal landmarks to localize the safe zone of the radial head, 24 different cadaveric elbows were internally fixed with a plate and screws through 3 different approaches (anterior, lateral, and posterolateral). Regardless of approach, the internal fixation allowed full forearm rotation in all the specimens. Utilization of this method and anatomic landmarks to localize the nonarticular portion of the radial head may assist the surgeon in open reduction and internal fixation of fractures of the radial head and neck. PMID- 9848564 TI - Revision of the ununited one-bone forearm. AB - Between 1975 and 1995, 7 patients were surgically treated for persistent forearm pain following previously unsuccessful attempts of constructing a radioulnar synostosis using interosseous bone grafting with either cross screw or pin fixation. These prior reconstructive procedures were salvage operations to address symptomatic radioulnar instability. The time interval between index operation and revision surgery was a mean of 12.6 months (range, 4-36 months). The postoperative follow-up period averaged 29.7 months (range, 11-61 months). To address these failed radioulnar arthrodeses, plate osteosynthesis and aggressive bone grafting were used in 5 of 7 patients (group 1) (4 with autologous graft and 1 with demineralized bone matrix). Union was achieved in all 5 patients after a single revision operation. Radiographic evidence of solid union was seen at approximately 4 months. In the remaining 2 patients (group 2), revision synostosis procedures were performed; these involved local bone grafting and repeat transverse screw fixation in 1 patient and iliac crest bone grafting alone in the other. One of these 2 patients progressed to union; the other had a persistent symptomatic fibrous union. The 5 patients in group 1 reported symptomatic relief, while the 2 patients in group 2 were dissatisfied. There were no postoperative complications in our series. Based on our findings, we advocate abundant bone grafting and plating for rigid internal fixation in failed radioulnar synostosis procedures. PMID- 9848565 TI - Acute dislocation of the trapeziometacarpal joint in a child. AB - An acute dorsal dislocation of the trapeziometacarpal joint in a child is presented. The mechanisms of injury and controversies of treatment are discussed. The achievement of a stable reduction acutely is emphasized. After healing, joint stability is confirmed by obtaining stress radiographs to identify whether joint subluxation is present before degenerative changes occur. PMID- 9848566 TI - Combined use of axillary block and lateral femoral cutaneous nerve block in upper extremity injuries requiring large skin grafts. AB - An axillary block and lateral femoral cutaneous nerve block combination was used in 11 patients with upper-extremity injuries requiring large skin grafts. In our experience, this block combination was highly successful. All but one of the patients obtained excellent pain relief (mean visual analog pain scores, 0.2) that required no narcotic analgesics or sedatives. There were no systemic or neurologic side effects attributed to the local anesthetic drugs. We believe that combining an axillary block and a lateral femoral cutaneous nerve block is a clinically useful and effective technique and an excellent anesthetic alternative for procedures requiring large grafts for the upper extremity. PMID- 9848567 TI - Late occlusion of microvascular vein grafts in replantation. AB - Two cases are described in which patients presented 16 and 17 years, respectively, after complete or incomplete amputation/replantation of the arm. In case 1, the patient complained of coldness, pain, and tingling in the replanted arm in the previous 24 hours and noticed that his fingers had gone white. Arteriography and subsequent surgery revealed obliteration of the vein graft (inserted in the distal brachial artery) by neointimal thickening and atherosclerotic plaque, which was confirmed in a subsequent morphologic examination. In case 2, the patient presented with discomfort and a pulsatile swelling on the inner aspect of his upper arm. Arteriography and surgery revealed an aneurysm in the previously inserted vein graft in the brachial artery, with some atherosclerotic degeneration. Both vein grafts were successfully replaced with a fresh autologous vein graft and the patients remain well several years later. The 2 cases suggest that as part of replantation surgery of a limb, it is essential to maintain postoperative clinical monitoring for signs of graft degeneration in all patients with long-term vein graft insertion. PMID- 9848568 TI - Recurrent histoplasmosis of the wrist: a case report. AB - Histoplasmosis of the extremities is rare. A case of recurrent histoplasmosis with a 10-year latency between initial presentation and clinical recurrence is reported. Prolonged antibiotic treatment and debridement of bony involvement led to clinical resolution of this fungal infection after a follow-up period of 20 months. PMID- 9848569 TI - Musculoskeletal outcome measures. PMID- 9848570 TI - Complications in internal fixation of the distal radius. PMID- 9848571 TI - Expanded profile of the SHAFT syndrome. PMID- 9848572 TI - Distal radius fractures: external fixation proves best. PMID- 9848573 TI - Pharmaceutical development of a parenteral lyophilized formulation of the investigational antitumor neuropeptide antagonist [Arg6, D-Trp7,9, MePhe8] Substance P [6-11]. AB - The aim of this study was to develop a stable parenteral dosage form for the investigational cytotoxic drug [Arg6, D-Trp79,MePhe8]-Substance P [6-11] (Substance P Antagonist G; Antagonist G). Antagonist G bulk drug was structurally and analytically characterized. The drug exhibits excellent aqueous solubility, although relatively poor aqueous stability characteristics. Lyophilization was, therefore, selected as the manufacturing process. Differential scanning calorimetry studies were conducted to determine the freeze-drying cycle parameters which resulted in a stable, lyophilized formulation of Antagonist G. The prototype, containing 50 mg Antagonist G per vial, was found to be the optimal formulation in terms of solubility, length of the freeze-drying cycle, stability, and dosage requirements in the planned phase I clinical trials. Quality control of the freeze-dried formulation showed that the manufacturing process does not change the integrity of Antagonist G. Shelf life studies demonstrated that the formulation is stable for at least 3 years, when stored at 2-8 degrees C in a dark environment. Oxidative degradation products of Antagonist G were isolated and structurally characterized by mass spectrometry, nuclear magnetic resonance spectroscopy, and infrared spectroscopy. PMID- 9848574 TI - Activity of pivaloyloxymethyl butyrate, a novel anticancer agent, on primary human tumor colony-forming units. AB - The anti-proliferative effects of pivaloyloxymethyl butyrate (AN-9), a butyric acid (BA) derivative with potent tumor-differentiating properties both in vitro and in vivo, was evaluated against colorectal, breast, lung, ovarian, renal cell, bladder, and other types of tumor colony-forming units in a human tumor cloning assay. A total of 76 evaluable specimens were exposed to AN-9 continuously, 48 of these were also exposed to BA continuously for direct comparison of the two agents, and 20 specimens were exposed to AN-9 for two hours. An in vitro inhibitory response was defined as a > or = 50% decrease in tumor colony formation in treated cells compared to untreated controls. Superior anti-tumor activity was observed with the continuous exposure to AN-9 (39% in vitro response at 100 microM and 70% at 200 microM) than with the two-hour exposure (20% at 100 microM and 25% at 200 microM). At a continuous concentration of 200 microM, AN-9 demonstrated greater tumor-specific activity than BA against melanoma (100% vs. 67%), ovarian (67% vs. 40%), breast (63% vs. 0%), non-small cell lung (60% vs. 10%), and colorectal tumor colony-forming units (62% vs. 20%). AN-9 is a novel differentiating agent with activity against colony-forming units derived from a variety of primary human tumors, including those that are considered relatively chemoresistant, and may thus provide a therapeutic alternative or addition to standard cytotoxic agents, if appropriate drug concentrations can be achieved in patients. PMID- 9848575 TI - Chronic effects of a novel synthetic anthracycline derivative (SM-5887) on normal heart and doxorubicin-induced cardiomyopathy in beagle dogs. AB - This study was designed to investigate the chronic cardiotoxic potential of SM 5887 and a possible deteriorating effect of SM-5887 on low-grade cardiotoxicity pre-induced by doxorubicin in beagle dogs. In the chronic treatment, beagle dogs of each sex were given intravenously once every 3 weeks, either a sublethal dose of doxorubicin (1.5 mg/kg) or SM-5887 (2.5 mg/kg). The experiment was terminated 3 weeks after the ninth dosing. Animals which received over six courses of doxorubicin demonstrated the electrocardiogram (ECG) changes, decrease of blood pressure and high-grade histopathological cardiomyopathy, while animals which were terminally sacrificed after the SM-5887 administration did not show any changes in ECG, blood pressure and histopathological examinations. To examine a possibly deteriorating cardiotoxic effect of SM-5887, low-grade cardiomyopathy was induced in dogs by four courses of doxorubicin (1.5 mg/kg). Nine weeks after pre-treatment, dogs were given four courses of either doxorubicin (1.5 mg/kg) or SM-5887 (2.5 mg/kg) once every 3 weeks. The low-grade cardiotoxic changes were enhanced by the additional doxorubicin treatment. On the contrary, the SM-5887 treatment did not progress the grade of cardiomyopathy. In conclusion, SM-5887 does not have any potential of chronic cardiotoxicity and deteriorating effect on doxorubicin-induced cardiotoxicity in dogs. PMID- 9848576 TI - Preclinical antitumor efficacy of analogs of XK469: sodium-(2-[4-(7-chloro-2 quinoxalinyloxy)phenoxy]propionate. AB - A series of quinoxaline analogs of the herbicide Assure was found to have selective cytotoxicity for solid tumors of mice in a disk-diffusion-soft-agar colony-formation-assay compared to L1210 leukemia. Four agents without selective cytotoxicity and 14 agents with selective cytotoxicity were evaluated in vivo for activity against a solid tumor. The four agents without selective cytotoxicity in the disk-assay were inactive in vivo (T/C > 42%). Thirteen of the fourteen agents with selectivity in the disk-assay were active in vivo (T/C < 42%). Five of the agents had curative activity. These five agents had a halogen (F, Cl, Br) in the 7-position (whereas Assure had a CI in the 6 position). All agents with curative activity were either a carboxylic acid, or a derivative thereof, whereas Assure is the ethyl ester of the carboxylic acid. All other structural features were identical between Assure and the curative agents. Assure had no selective cytotoxicity for solid tumors in the disk-assay, and was devoid of antitumor activity. The analog XK469 is in clinical development. PMID- 9848577 TI - Gemcitabine plus dose-escalated epirubicin in advanced breast cancer: results of a phase I study. AB - Gemcitabine has shown single-agent activity in metastatic breast cancer. Epirubicin is also widely used for the adjuvant and treatment of metastatic breast cancer. The toxicity profiles and modes of action are different which provides a good rationale for studying both drugs in combination. In a phase I study gemcitabine at a fixed dose of 1000 mg/m2 on days 1, 8, 15 of a 28 day cycle was combined with escalated weekly doses of epirubicin starting with an initial dose of 10 mg/m2. Patients had stage IV metastatic disease without previous chemotherapy except as adjuvant treatment. Nineteen patients were included in the study which defined the maximum tolerated dose (MTD) of epirubicin at 20 mg/m2. Myelosuppression was the dose limiting toxicity with leucopenia WHO grade 3 and 4 in 40.0% and 20.0%, neutropenia WHO grade 3 and 4 without neutropenic fever in 20.0% and 40.0% and thrombocytopenia WHO grade 4 in 20.0%. At the epirubicin 15 mg/m2 dose level, leucopenia (11.1% WHO grade 3) and neutropenia (12.5 and 37.5% WHO grade 3 and 4) were reported. Symptomatic toxicity was generally mild: nausea/vomiting in about 20% of patients (WHO grade 3 or 4) on both 15 and 20 mg/m2 epirubicin dose levels. Alopecia WHO grade 3 and 4 was seen in 2 patients at MTD. Four of 19 evaluable patients had a partial response. We conclude that the combination of gemcitabine and epirubicin is well tolerated and has promising activity. A phase II study is underway with gemcitabine 1000 mg/m2 and epirubicin 15 mg/m2 on days 1, 8 and 15 of a 28 day cycle. PMID- 9848578 TI - Phase I and pharmacokinetic study of intraperitoneal topotecan. AB - OBJECTIVE: To determine the maximum tolerated dose and pharmacokinetics of topotecan when administered by the intraperitoneal route. METHODS: A dose escalating Phase I trial was conducted in which fifteen % of the total dose was given as an intraperitoneal bolus in two litres of D5W and the remainder was given as a continuous intraperitoneal infusion over 24 hours. Treatments were given every 21 days. Pharmacokinetic analyses were performed at the recommended phase II dose. RESULTS: Seventeen patients received a total of 43 cycles at 21 day intervals. The maximum tolerated dose was 4 mg/m2 and acute dose-limiting toxicity was neutropenia. Other toxicities included leukopenia, anemia, emesis, fever, and abdominal pain. Although no objective responses were achieved, five of ten patients with ascites had a decrease in fluid accumulation with administration of intraperitoneal topotecan. The recommended phase II dose is 3 mg/m2. Pharmacokinetic analysis performed at a dose of 3 mg/m2 demonstrated that elimination from the peritoneal cavity followed second-order kinetics with k1 = 1.6 hr(-1), k2 = 0.3 hr(-1) and first and second-phase half-lives of 0.49 and 2.7 hours, respectively. Plasma pharmacokinetic behavior was best described by first order kinetics with k = 0.5 hr(-1) and a half-life of 3.9 hours. The pharmacologic advantage, expressed as the peritoneal to plasma AUC ratio was 31.2. CONCLUSIONS: Intraperitoneal administration of topotecan at 3 mg/m2 results in a substantial increase in drug exposure for the peritoneal cavity without compromising systemic exposure; this may be beneficial for the treatment of patients with ovarian cancer or intraperitoneal carcinomatosis. PMID- 9848579 TI - Phase I/II evaluation of pentostatin (2'-deoxycoformycin) in a five day schedule for the treatment of relapsed/refractory B-cell chronic lymphocytic leukaemia. AB - The purpose of this study was to determine the efficacy and toxicity of pentostatin (2'-deoxycoformycin) administered in a five day schedule every 28 days to patients with B-cell chronic lymphocytic leukaemia (B-CLL) relapsed from or refractory to at least one line of prior chemotherapy. The initial dose level of 2 mg/m2/day was adjusted up or down by 0.5 mg/m2 in subsequent cycles on the basis of haematological and non-haematological toxicities. The five day schedule was selected because published pharmacokinetic studies had indicated that although pentostatin had an elimination half-life of approximately six hours and could inhibit plasma adenosine deaminase activity for 24 hours, recovery of enzyme activity rapidly took place and accumulation of dATP which has a toxic effect on non-replicating lymphoid cells could be increased by repeated dosing. Twenty-nine patients were entered into the study and dose-escalation was possible in nine, while dose reductions were required for five patients. Of the 24 patients evaluable for response, complete responses were achieved in two and partial responses in five for an overall response rate of 29.2%. Toxicity consisted of myelosuppression, infection, nausea and vomiting and hepatotoxicity but was experienced at acceptable levels considering the heavily pre-treated nature of the patient population. Pentostatin in this schedule has salvage activity in previously treated or resistant patients with B-CLL. PMID- 9848580 TI - A phase I clinical trial of prolonged infusion of hydroxyurea in combination with hyperfractionated, accelerated, external radiation therapy in patients with advanced squamous cell cancer of the head and neck. AB - BACKGROUND: Preclinical data suggested that sustained inhibition of the anabolic enzyme, ribonucleotide reductase (RR), by hydroxyurea (HU) may be critical for the anticancer effects of the drug. A phase I trial of continuous infusion HU with concomitant hyperfractionated, accelerated radiation therapy (CHU-CHRT) was initiated to determine the maximum tolerated dose (MTD) and dose limiting toxicities (DLT) of HU in patients with locally advanced squamous cell carcinoma (SCC) of the head and neck. METHODS: Patients were required to have histologically-documented and radiographically-staged locally advanced SCC of the hypopharynx (AJC stages II, III or IV), oropharynx (AJC stage IV), or oral cavity (AJC stage IV) not amenable to reasonable surgical resection. Eligible patients had adequate bone marrow, hepatic, and renal function and had to give informed consent. Concomitant, hyperfractionated, accelerated radiation therapy (CHRT) consisted of 1.2 Gy BID (6 hour minimum interfraction interval) on weekdays and 1.2 Gy delivered daily on the weekends to a total tumor dose of 74.4 Gy. Continuous infusion hydroxyurea (CHU) was administered at 0.25-0.375 mg/m2/min as a continuous intravenous infusion daily for 5 days with weekends days off for the duration of the radiation therapy. The dose of HU was increased by 0.125 mg/m2/min between dose levels until DLT was reached in 2/6 patients. If the primary had a complete clinical response and biopsies were negative, planned neck dissections were performed. RESULTS: Fifteen patients were enrolled and are evaluable. The initial dose level, 0.25 mg/m2/min was tolerated by 3/3 patients. At 0.375 mg/m2/min, 3/6 patients experienced grade 3-4 infections, with one patient having a non-fatal, subendocardial infarction. At 0.313 mg/m2/min, no patient experienced DLT. CONCLUSION: The MTD for CHU-CHRT was 0.313 mg/m2/min. The toxicities were primarily mucosal and a phase II study is in progress. PMID- 9848581 TI - Phase II trial of oral beta-all trans-retinoic acid in hepatocellular carcinoma (SWOG 9157). AB - Twenty-nine chemotherapy-naive patients with primary hepatocellular carcinoma were treated with oral beta-all trans-retinoic acid (retinoic acid, TRA 50 mg/m2 t.i.d.) on a 3-week on/one week off schedule until progression or grade 3 or 4 toxicity. Eligibility requirements allowed abnormal liver function tests as long as the creatinine and bilirubin levels were normal. No responses were seen and the median survival was four months. Grade 3 side effects occurred in II patients and grade 4 in four and included a wide range of toxicities. The results indicate that oral TRA is ineffective against primary hepatocellular carcinoma and suggest that dose-modification of this retinoid may be required in patients with significant malignant hepatic involvement. PMID- 9848582 TI - Phase II study of prolonged infusion of Taxol in patients with metastatic colorectal carcinoma. AB - PURPOSE: Taxol represents a new class of anticancer agents with activity against a wide variety of solid tumors. In preclinical systems, its cytotoxicity is schedule dependent with prolonged exposure being more effective. We studied a 120 hour infusion schedule of Taxol in patients with metastatic measurable colorectal carcinoma who had had one prior 5-FU-based chemotherapy. METHODS: Patients with measurable metastatic colorectal carcinoma were eligible. Patients had to have normal liver, renal, and bone marrow functions. Written informed consent was obtained from all patients. The starting dose of Taxol was 150 mg/m2 infused over 120-hours in the outpatient setting. Taxol was repeated every 21 days. RESULTS: Fifteen patients were registered. Among 14 evaluable patients, we did not observe any complete or partial response. Major toxicity included myelosuppression and mucositis. There was no treatment-related death. CONCLUSION: Taxol administered by this schedule was found ineffective in patients with metastatic colorectal carcinoma who had previously received one 5-FU-based chemotherapy. PMID- 9848583 TI - Phase II trial of pyrazine diazohydroxide in androgen-independent prostate cancer. AB - No effective therapy has been demonstrated for hormone refractory prostate cancer (HRPC). Pyrazine diazohydroxide (PZDH) is a novel antineoplastic agent with a broad range of activity in preclinical studies and a moderate toxicity profile in Phase I trials. We undertook a Phase II study of PZDH in HRPC utilizing decline in PSA as the primary end point. Fifteen patients were enrolled, median age of 70 (55-86), median pretherapy PSA 206 ng/ml (range 42-10,000). Four patients were African American. Sites of disease: bone only 7, soft tissue only 2, both 6. All were evaluable for toxicity and response. PZDH was administered at 250 mg/m2 i.v. every three weeks. The median number of cycles administered was two (range 1-6). Toxicity was mild, with only one patient manifesting serious (grade 3-4) toxicity. Unfortunately, activity was minimal with only a single patient demonstrating a >75% decline in PSA. As this patient's PSA began to rise almost immediately the response was considered transient and not felt to justify pursuing a second stage of the trial. Supporting this conclusion was the disappointing median survival of 220 days. In summary, we conclude that PZDH, while well tolerated at this dose and schedule has only minimal activity in HRPC. PMID- 9848584 TI - Phase II study of paclitaxel (BMS-181339) intravenously infused over 3 hours for advanced or metastatic breast cancer in Japan. BMS-181339 Breast Cancer Study Group. AB - A Phase II study of paclitaxel in patients with primary advanced or metastatic breast cancer was conducted by a cooperative study group consisting of 16 institutions in Japan. Paclitaxel at a dose of 210 mg/m2 was intravenously infused over 3 hours, along with premedication to prevent hypersensitivity reactions. The course was repeated at 21-day intervals. Of 62 eligible patients, 60 were evaluable for toxicity and 59 were evaluable for efficacy. Forty-five patients were previously treated with anthracyclines. Twenty-one of 59 patients (35.6%) had a major objective response including 2 CRs and 19 PRs (95% confidence interval, 23.6-49.1%). A response rate of 35.5% (CR1, PR10) was observed in 31 patients refractory to the anthracyclines containing prior metastatic chemotherapy. Median (range) time was 41 (6-100) days to onset of and median duration of response was 125 (36-305) days. Toxicities included leukopenia (grade 3, 4: 67%), anemia (grade 1-3: 80%), thrombocytopenia (grade 1: 8%), alopecia (grade 3: 43%), peripheral neuropathy (grade 1-3: 93%), arthralgia (59%), myalgia (46%), nausea and vomiting (40%), fever (33%), allergic reaction (grade 3: 2%) and hypotension (grade 3: 5%). All toxicities were tolerable and manageable. Paclitaxel intravenously infused over 3 hours demonstrated a significant antitumor activity for metastatic breast cancer. Furthermore, paclitaxel exhibited non-cross resistance to anthracycline. Paclitaxel administered as a convenient 3-hour infusion is effective for patients with metastatic breast cancer and has an acceptable toxicity profile. PMID- 9848586 TI - Complications in arthroscopic shoulder surgery. AB - Arthroscopic surgery of the shoulder is considered a safe technique, yet a variety of complications have been described. There are few published reports on the incidence of these complications. The aim of this study is to evaluate the complication rate of arthroscopic shoulder surgery. We retrospectively evaluated a series of 179 consecutive arthroscopic (n=141) and combined (arthroscopic plus open; n=38) procedures performed by the same surgeon. The overall complication rate was 9.49%. Combined procedures had a 5.26% complication rate and arthroscopic procedures a 10.63% complication rate. PMID- 9848585 TI - Topotecan in previously treated advanced urothelial carcinoma: an ECOG phase II trial. AB - BACKGROUND: [corrected] Chemotherapeutic agents are playing an increasing role in the management of urothelial carcinoma. Despite recent advances in the treatment of this disease there continues to be a need to identify new active agents and their toxicity spectra. Topotecan is an agent as yet unstudied in bladder cancer. METHODS: Ambulatory patients with progressive advanced urothelial carcinoma following prior systemic chemotherapy were treated with topotecan 1.5 mg/m2 intravenously (i.v.) daily for 5 days every three weeks for 6 cycles. Doses were modified for leukopenic fever, thrombocytopenic bleeding, and any grade 3 or 4 (NCI common toxicity criteria) toxicity. RESULTS: Forty-four eligible patients entered the trial. There were 4 partial responses for an overall response rate of 9.1% (exact 95% two-stage binomial CI, 2.9% to 25.5%). Major identified toxicities were gastrointestinal and myelosuppression. There were no treatment related deaths. CONCLUSIONS: Topotecan at this dose and schedule has minimal activity in previously treated patients with advanced urothelial carcinoma. Toxicities can be severe but are manageable. PMID- 9848587 TI - Specificity of the Speed's test: arthroscopic technique for evaluating the biceps tendon at the level of the bicipital groove. AB - A positive Speed's test result is usually thought to suggest inflammation or lesions related to the biceps/labral complex. The specificity, sensitivity, and positive and negative predictive values are determined for the Speed's test. A prospective study design was developed for all patients with shoulder pain who presented between October 1, 1994 and February 28, 1995. The clinical results of the Speed's test were correlated with biceps/labral pathology by direct arthroscopic visualization. A neuroprobe is used to pull the biceps tendon into the articular portion of the glenohumeral joint so as to visualize the biceps tendon at the level of the bicipital groove. Forty-six shoulders in 45 patients, 31 men (average age, 53 years; range, 16 to 76 years) and 14 women (average age, 64 years; range, 30 to 80 years) with 26 dominant and 20 nondominant extremities were operated on during this time interval. The clinical evaluation showed that the speed's test was positive in 40 shoulders. Biceps/labral complex pathology was present in 10 of these patients. A specificity of 13.8%, a sensitivity of 90%, a positive predictive value of 23%, and a negative predictive value of 83% were calculated. Thus, it is concluded that the Speed's test is a nonspecific but sensitive test for macroscopic biceps/labral pathology. This clinical examination is positive with a various number of other pathological shoulder problems. PMID- 9848588 TI - The arthroscopic management of osteochondritis dissecans of the adolescent elbow. AB - The management of avascular necrosis of the capitellum of the adolescent elbow continues to be a dilemma. This article is a critical retrospective analysis of 12 pediatric patients (mean age at surgery 14.5 years) who underwent arthroscopic debridement alone followed by early range of motion. Follow-up at a mean of 3.2 years (range, 2.0 to 5.7 years) indicated that the average flexion contracture improved from 23 degrees preoperatively to 10 degrees postoperatively. All patients had remodeling of the capitellum by plain radiographs; however, five patients had associated enlargement of the radial head. Eleven patients had minimal mechanical symptoms after the procedure and were highly satisfied. One patient had substantial enlargement of the radial head with continued loss of supination and mechanical symptoms requiring radial head resection 2 years after the index procedure. Five patients had a triangular avulsion fragment present off the lateral capsule. A statistically significant worse subjective outcome was associated with the presence of this fragment (P < .005). There were no complications. PMID- 9848589 TI - Patellar tendon defect during the first year after anterior cruciate ligament reconstruction: appearance on serial magnetic resonance imaging. AB - The purpose of this study was to use magnetic resonance imaging to evaluate various parameters of the patellar tendon during the first year after harvest for anterior cruciate ligament (ACL) reconstruction. Twelve consecutive patients were serially imaged on a 1.5 Tesla GE magnet (GE Medical Systems, Milwaukee, WI) with a dedicated knee coil at 3 weeks, 3 months, 6 months, and 1 year after undergoing ACL reconstruction using a central one-third patellar tendon autograft. The tendon defect was not closed primarily, but the paratenon was approximated. The following measurements were performed: tendon width, defect width, cross sectional area of the tendon, and tendon length. In addition, the patellar bone harvest site was evaluated for healing. The width of the tendon defect decreased by 62% over 12 months (P < .05). Only two patients showed complete closure of the defect. Tendon width was noted to decrease by 6.5% (P=.017). The ratio of defect width to overall tendon width (designated R) decreased by 58% (P < .05). Tendon length was noted to decrease during this by 8% (P=.037). The tendon cross sectional area was noted to increase by 9% at 1 year, but this was not found to be statistically significant (P=.39). One year after ACL reconstruction using a central one-third patellar tendon, the tendon defect has begun to reconstitute itself but there is still a significant gap. This persistent defect must be taken into consideration when planning revision ACL surgery using reharvest of the central one third of the patellar tendon. The entire tendon also exhibits a reduction in width and length, while cross-sectional area increases slightly. Complete healing of the graft defect can not be assumed at 12 months post-ACL reconstruction. PMID- 9848590 TI - Bone tunnel enlargement after anterior cruciate ligament reconstruction with semitendinosus tendon using Endobutton fixation on the femoral side. AB - We evaluated 29 knees with a minimum follow-up of 2 years after anterior cruciate ligament (ACL) reconstruction using doubled autogenous semitendinosus tendons. On the femoral side, a 5-mm Mersilene tape (Ethicon, Norderstedt, Germany) with an Endobutton (Acufex Microsurgical, Mansfield, MA) was used. The tendon was fixed on the tibial side with two staples. Regarding the IKDC score, 66% of the patients were graded as normal or nearly normal. The anterior laxity side-to-side difference (KT 1000, man-max-drawer) was under 3 mm in 55% and under 5 mm in 90%. Radiographs taken in the lateral and anteroposterior projections of the knee showed sclerotic bone tunnel margins. The diameter of the bone tunnels were measured, corrected for magnification, then compared with the original reamed diameter to determine any change in size. Enlargement of at least 2 mm was identified in 72% of the femoral tunnels and 38% of the tibial tunnels. No correlation was found concerning the enlargement of the tunnel and the IKDC score or the residual joint laxity. We conclude that using an Endobutton-Mersilene construct in ACL reconstruction leads to femoral and tibial bone tunnel enlargement at follow-up of 2 years. PMID- 9848591 TI - A comparison of ketorolac tromethamine/oxycodone versus patient-controlled analgesia with morphine in anterior cruciate ligament reconstruction patients. AB - Effective postoperative analgesia with minimal side effects remains an important goal in enabling increasingly complex surgical procedures to be performed on an outpatient basis. In this study, we examined the efficacy of postoperative analgesia in 90 patients undergoing anterior cruciate ligament reconstruction using a patellar tendon autograft, with a 24-hour hospital stay. Patients were randomized to receive either intramuscular ketorolac supplemented by oral oxycodone, or intravenous morphine via patient-controlled analgesia (PCA) device, for postoperative analgesia. Patients were monitored for 2 hours in the recovery room, then every 4 hours until discharge, for the presence of complications of nausea, vomiting, urinary retention, pruritus, and dizziness. Pain was assessed using a visual analog scale (VAS) on the morning of postoperative day one. All patients were discharged by 24 hours after surgery. Ten (20%) of the patients receiving ketorolac/oxycodone versus 31 (79%) of those receiving PCA morphine experienced postoperative complications (P < .05). Postoperative nausea, vomiting, and urinary retention were each significantly more common in the PCA morphine group (P < .05). The incidence of pruritus and dizziness was low overall. There was no significant difference between groups in the severity of postoperative pain as assessed using a VAS. We conclude that ketorolac/oxycodone may provide comparable analgesia with fewer undesirable side effects than PCA morphine in patients undergoing anterior cruciate ligament reconstruction. Patients receiving ketorolac/oxymorphone may have a better quality recovery and more rapid discharge. PMID- 9848592 TI - Identification of the vascular and avascular zones of the human meniscus using magnetic resonance imaging: correlation with histology. AB - Since the initial employment of magnetic resonance imaging (MRI) to diagnose meniscal tears, a characteristic low-signal intensity, triangular-shaped structure has been interpreted as representing the entire meniscus. The difficulty in diagnosing meniscocapsular separations with MRI has brought attention to our lack of understanding of the appearance on MRI of the outer third of the meniscus and the meniscocapsular junction. We correlated MRIs of the meniscus in cadaver knees with histological sections and found that the low signal, wedge-shaped structure corresponds only to the avascular (white) zone of the meniscus, whereas the high-signal zone peripheral to it corresponds to the vascularized (red) zone. PMID- 9848593 TI - Clinical results of meniscus repair in patients 40 years and older. AB - The critical role that the meniscus plays in the knee along with the advantages of preserving as much of the meniscus as possible have both been well documented. Whenever possible, meniscus repair has become the procedure of choice for treatment of meniscal tears. However, some researchers have reported less favorable results in older patients. To determine the results of meniscus repair in older patients, patients 40 years and older who underwent arthroscopically assisted meniscus repair were prospectively followed up. Thirty-seven patients were included in the study, all of which had a minimum 2-year follow-up (average, 26.5 months). The average age of the patients at the time of the repair was 44.2 years (range, 40 to 52 years); 26 were males and 11 were females. There were 19 left knees and 18 right knees included in the study. Twenty-two patients had associated anterior cruciate ligament reconstructions. Physical examination at follow-up included swelling, joint line tenderness, locking, and McMurray testing and radiographs. Five of 37 menisci repaired (13.5%) were symptomatic at latest follow-up. All of these patients had joint line tenderness and two had a positive McMurray test. Three of these patients had repeat arthroscopy confirming that the meniscus had not healed. Because of the small number of patients with symptoms at follow-up, the authors feel that meniscus repair in patients 40 years and older is an effective treatment for peripherally located meniscus tears. With 86.5% of the patients having good clinical results, these findings are comparable with other studies with a younger population and signify that repair of peripheral tears is indicated in this age group especially in conjunction with anterior cruciate ligament reconstruction. The findings suggest that the location and meniscal tear, rather than the age of the patient, determine the potential for successful repair. PMID- 9848594 TI - The clinical significance of the complete type of suprapatellar membrane. AB - The plicae are synovial septa remaining in adult life that existed in early fetal life. The suprapatellar plica separates the suprapatellar pouch from the knee joint. The plica syndrome has clinical significance, which has been occasionally overlooked, but the pathophysiology of symptomatic plicae may be hard to explain. To evaluate the clinical significance of the suprapatellar plicae, the authors reviewed 34 cases in 23 patients with vague pain around the knee and a total septum of the suprapatellar plica at arthroscopic examination from September 1991 to December 1993. The follow-up period was from 6 months to 2 years and 9 months. The most common presenting symptom was chronic mild knee pain, aggravated by prolonged squatting or standing, with other patients reporting recurrent swelling, instability, giving-way, or a feeling of snapping. The objective findings include palpable band on the superomedial side, audible snapping, and local tenderness, but there were no significant abnormal findings in the laboratory. The radiographic findings were few, with sclerosis of the articular surface of the patella in 2(6%), malalignment in 1(3%), and mild degenerative change in 4 cases(12%). Five of 11 patients evaluated by bone scan had increased uptake around the patellofemoral joint, and 7 of 13 knees had a relatively small suprapatellar bursa on conventional arthrogram or pneumoarthrogram. At arthroscopy, a suprapatellar plicae with complete septum was identified in 30 of 34 cases (88%) and associated lesions presented as meniscal tears, loose body, and discoid meniscus without tear. The complete plicae were surgically excised under arthroscopic control in 30 patients and the results were excellent in 22 patients (73%), good in 5 (17%), and poor in 3 (10%)at 17 months follow-up; there were no failures. In our opinion, the complete suprapatellar plica is clinically significant in patients who have equivocal diagnosis of knee problems and further studies of the pathophysiology of complete suprapatellar plica are needed. PMID- 9848595 TI - Operative treatment and arthroscopic findings in chronic patellar tendinitis. AB - Patellar tendinitis is a well-described clinical entity that usually responds well to conservative treatment. However, a subset of patients continue with symptoms despite exhaustive nonoperative means. The objective of the study was to review the treatment course and operative results of seven patients (eight knees) with chronic patellar tendinitis treated surgically. Five (70%) of these patients were either professional or collegiate athletes. The average age was 30 years (range, 20 to 45 years). Duration of symptoms averaged 1.4 years (range, 3 months to 4 years) before surgical correction. Operative treatment included knee arthroscopy and open repair consisting of excision of degenerated tissue and stimulation of a healing response of the patellar tendon pathology. Operative findings and pathology reports consistently showed marked fibrotendinous degeneration. Follow-up averaged 3.6 years (range, 4 months to 8.5 years). Outcomes were measured subjectively with SF36 results and objectively with Biodex testing (Biodex, Shirley, NY) and return to previous level of competition. Overall, 86% of patients achieved an excellent result and 14% had a fair result. We recommend operative intervention in a patient with chronic patellar tendinitis who does not improve with well-supervised, comprehensive conservative treatment. PMID- 9848596 TI - Overuse tendon conditions: time to change a confusing terminology. AB - In overuse clinical conditions in and around tendons, frank inflammation is infrequent, and is associated mostly with tendon ruptures. Tendinosis implies tendon degeneration without clinical or histological signs of intratendinous inflammation, and is not necessarily symptomatic. Patients undergoing an operation for Achilles tendinopathy show similar areas of degeneration. When the term tendinitis is used in a clinical context, it does not refer to a specific histopathological entity. However, tendinitis is commonly used for conditions that are truly tendinoses, and this leads athletes and coaches to underestimate the proven chronicity of the condition. Paratenonitis is characterized by acute edema and hypermia of the paratenon, with infiltration of inflammatory cells, possibly with production of a fibrinous exudate that fills the tendon sheath, causing the typical crepitus that can be felt on clinical examination. The term partial tear of a tendon should describe a macroscopically evident subcutaneous partial tear of a tendon, an uncommon acute lesion. Most articles describing the surgical treatment of 'partial tears' of a given tendon in reality deal with degenerative tendinopathies. The combination of pain, swelling, and impaired performance should be labeled tendinopathy. According to the tissues affected, the terms tendinopathy, paratendinopathy, or pantendinopathy should be used. PMID- 9848597 TI - The effect of different graft tensioning in anterior cruciate ligament reconstruction: a prospective randomized study. AB - A prospective study was performed to establish the influence on stability of tensioning the graft after anterior cruciate ligament reconstruction with a bone patellar tendon-bone graft. There were 38 consecutive patients randomly divided into two groups; in the first group (19 patients) the graft was tensioned at 20 N, and in the second group the tension was 40 N. Evaluation included Lysholm scores, instrumented Lachman tests, IKDC level, and lateral radiographs to measure the tibia position versus the femur. One year after surgery we did not find any significant difference between the two groups. We conclude that the importance of graft tensioning has yet to be established in the clinical situation. Based on our results, graft tension of 20 N seems to be sufficient, without the risk of overconstraining the knee joint. PMID- 9848598 TI - Arthroscopic surgery of the metatarsophalangeal first joint. AB - Arthroscopic surgery of the metatarsophalangeal first joint was used to treat a dorsal impingement syndrome of the hallux by removing the dorsally located osteophytes. Also, osteochondritis dissecans, painful sesamoid bones resistant to conservative therapy, and hallux rigidus were arthroscopically treated in a prospective study enrolling 24 consecutive patients. Seventeen patients were high level athletes. Pain, swelling, sports, and work involvement were recorded. The operative technique is described. There was one persistent loss of sensitivity of the hallux. In the dorsal impingement group, 8 of 12 patients had a good or excellent result after a minimum follow-up of 2 years. In patients with osteochondritis dissecans, 3 of 4 showed good or excellent results. In hallux rigidus and sesamoid bone removal the results were less favorable. As in all arthroscopic procedures there was a fast rehabilitation and work resumption. Especially in athletes arthroscopically, treatment of a dorsal impingement syndrome or osteochondritis dissecans can be recommended because of the diminished chance of scar fibrosis. PMID- 9848599 TI - SLAP lesions: a retrospective multicenter study. AB - A retrospective analysis of 530 glenohumeral arthroscopies performed by three independent Belgian arthroscopists revealed the presence of 32 SLAP lesions, which represents an incidence of 6%. Since this is exactly the same percentage as found by Snyder et al., we report our data in this article. We classified 23 of the SLAP lesions using Snyder's classification, 7 needed the additional classification of Maffet et al., and 2 lesions were considered to be anatomic variations; 53% of the lesions were of type II. Concerning the mechanism of injury, we found comparable percentages of traction (22%) and compression (28%) injury as reported by Snyder, but also a high number (25%) of overhead sports activities as described by Andrews et al. Associated lesions were in close accordance with Snyder's data, but a relatively low incidence of rotator cuff injuries (10%) was present. Comparison of treatment regimens showed that the same percentage of lesions (34%) was fixed arthroscopically in both series. Only SLAP II, IV, and V lesions must be considered as unstable and in need of fixation. We confirm that patients' complaints and clinical symptoms are vague and inconsistent. Imaging, using computed tomographic arthrography or magnetic resonance, was performed in a minority of cases. Advantages and pitfalls of both techniques are discussed. Anatomic variations causing an extra-large sublabral hole are shown, and we warn about potential diagnostic and therapeutic errors in these cases. PMID- 9848600 TI - A known technique for meniscal repair in common practice. AB - In this retrospective study, we calculated the healing rate of meniscal repairs performed with an outside-in technique. We describe complications encountered and evaluate some known criteria used in the decision to perform a meniscal repair instead of partial meniscectomy. Included is a brief description of the surgical technique and of the trauma type and the meniscal lesions that were repaired. The technique has a high degree of success (74% of the meniscal repairs survived during a mean follow-up of 3.5 years). Although there is a 25% complication rate, no serious or permanent complications were added by repairing menisci instead of performing a partial meniscectomy. For this reason we think saving even a relatively low percentage of menisci may be worthwhile. We can also conclude that an anterior cruciate ligament-deficient knee that is stable can still have a good result in meniscal repair, without performing cruciate reconstruction. In each case, however, individual patient issues such as age, activity level, and associated lesions have to be considered. PMID- 9848601 TI - Different aspects of the cyclops lesion following anterior cruciate ligament reconstruction: a multifactorial etiopathogenesis. AB - After anterior cruciate ligament (ACL) reconstruction using a patellar-tendon autograft, 65 patients underwent second-look arthroscopy in conjunction with hardware removal. In 23 patients, hypertrophic tissue was found in the anterior part of the knee. This tissue presented different aspects, from a well synovialized nodule to a more disorganized fibrous tissue according to patients' complaints. Endoscopic resection of this offending tissue was generally sufficient to obtain a satisfactory result. In patients presenting a loss of extension, the notch frequently had to be enlarged. We have found a multifactorial pathogenesis to be likely: the nodule is a natural fibroproliferative tissue process originating either from drilling debris from the tibial tunnel or from remnants of the ACL stump and, more rarely, from broken graft fibers. Sometimes, when the graft is malpositioned, the scar tissue can result from repeated graft impingement on the notch at terminal extension. Formation of this aberrant tissue should be prevented by proper positioning of the graft, by enlargement of the narrowed notch in chronic cases, by using drills of increasing diameters to avoid production of osteocartilaginous fragments, by meticulous resection of all drilling debris and ACL remnants around the tibial tunnel, and by enlarging the notch roof if any contact with the graft is present when the knee is fully extended intraoperatively. PMID- 9848602 TI - Limited value of arthroscopic evaluation and treatment of painful knee prostheses: a retrospective study of 27 cases. AB - Twenty-seven painful knee replacements were evaluated arthroscopically. The diagnostic and therapeutic value of these arthroscopic procedures was studied retrospectively. In 5 of the 27 cases, the arthroscopy revealed no diagnosis for the pain. Some form of arthroscopic treatment was performed in 20 cases; in 6 of these 20 cases, however, the treatment did not reduce the pain. Based on these findings, we conclude that the indications for arthroscopic evaluation and treatment of painful knee prostheses are limited. PMID- 9848603 TI - Bursoscopy of the trochanteric bursa. AB - We report two cases in a single patient with severe, refractory trochanteric bursitis, treated successfully with bursoscopy and bursectomy. The symptoms, clinical findings, surgical technique, and short-term results are described. PMID- 9848604 TI - MDR-TB and the developing world--a problem no longer to be ignored: the WHO announces 'DOTS Plus' strategy. PMID- 9848605 TI - Air pollution in urban areas: the role of automotive emissions as a public health problem. PMID- 9848606 TI - The dilemma of MDR-TB in the global era. PMID- 9848607 TI - Outcome of chemotherapy in 107 patients with pulmonary tuberculosis resistant to isoniazid and rifampin. AB - SETTING: National Masan Tuberculosis Hospital, Korea. OBJECTIVE: Treatment for multidrug-resistant tuberculosis (MDR-TB) is considered to be clinically important, but there are few reports on this topic. We therefore retrospectively evaluated the outcomes of chemotherapy only for pulmonary MDR-TB. DESIGN: We reviewed the clinical courses of 107 patients with pulmonary disease due to Mycobacterium tuberculosis resistant to rifampin and isoniazid who were under follow-up between March 1996 and June 1996 after hospitalization between January 1993 and January 1996. We performed a retrospective cohort study for all the patients' records. Their regimens were selected individually and preferably included four medications that they had not been given previously and to which the strain was fully susceptible. RESULTS: The 107 patients (mean age 38.3 years) had previously received a mean of five drugs, and were shedding bacilli that were resistant to a mean of four drugs. Of 63 patients with sufficient follow-up data, 52 (82.5%) responded to chemotherapy (as indicated by negative sputum cultures for at least three consecutive months); 11 (17.5%) had no response, as shown by continually positive cultures. In a univariate analysis, an unfavorable response was significantly associated with resistance to a greater number of drugs before the current courses of treatment (relative risk 21.5; 95% confidence interval 1.2 3.0; P < 0.05). The mean period of follow-up was 17 months. There was no subsequent relapse among the patients with responses, and there were no tuberculosis-related deaths. CONCLUSION: In this study, multidrug-resistant pulmonary tuberculosis responded relatively well to carefully selected regimens. PMID- 9848608 TI - The effect of prior BCG vaccination on the clinical and radiographic presentation of tuberculosis meningitis in children in Istanbul, Turkey. AB - SETTING: Department of Pediatrics, Istanbul University Faculty of Medicine, Istanbul, Turkey, between January 1984 and December 1996. OBJECTIVE: To explore whether neonatal BCG vaccination offers any protective effect on clinical and laboratory profiles of central nervous system (CNS) tuberculosis in children. DESIGN: A retrospective review of cases of CNS tuberculosis diagnosed and treated in one institution. RESULTS: Of the 145 patients identified with CNS tuberculosis, 9.6% were vaccinated during the neonatal period. The rate of close contact with contagious pulmonary tuberculosis in family, age distribution, clinical findings and laboratory investigations on admission were not significantly different in vaccinated and non-vaccinated children. Although mortality rate in the vaccinated patients (8.3%) was found to be nearly half of that in the non-vaccinated group (15.1%), severe sequelae were significantly more frequent (P < 0.02) among the vaccinated patients; CONCLUSION: This study shows that neonatal BCG vaccine has little effect on the clinical findings of subsequent CNS tuberculosis, and that these children have typical presentations of tuberculosis disease. PMID- 9848609 TI - Factors determining compliance with tuberculosis treatment in an urban environment, Tamatave, Madagascar. AB - SETTING: Patient compliance remains one of the main obstacles that need to be overcome by tuberculosis control programmes, in the developing world as well as in industrialised countries. A better understanding of the various factors accounting for treatment default could help to achieve better compliance from patients. OBJECTIVES: To increase the understanding of the determining factors of default in an urban environment where medical facilities are accessible. Different kinds of determinants were studied: objective socio-demographic factors, subjective psychological factors, attitudes and behaviour of patients in relation to chemotherapy, quality of relationships with the medical staff, knowledge and attitudes regarding tuberculosis. DESIGN: Risk factors for default were assessed by a retrospective case-control study in a sample of 38 patients who had not completed treatment for pulmonary tuberculosis, compared with 111 controls who had completed treatment under comparable conditions. RESULTS: Default appears to be significantly linked to transportation time, the sex of the patient, patient information and the quality of communication between patients and health workers. False addresses given by patients are both a methodological bias and a risk factor for future default. CONCLUSION: Improved communication skills and attention from the medical staff could encourage more patients to complete their TB treatment. PMID- 9848610 TI - Tuberculosis among health care workers in British Columbia. AB - OBJECTIVES: To compare the clinical features and prevalence of active TB in British Columbia (BC) health-care workers (HCWs) with those of the general population, between 1991 and 1996. METHODS: Comparison of 25 HCWs and 50 controls randomly selected from the Centres for Disease Control registry, with respect to demographics, prevention, diagnosis and management. RESULTS: HCWs had fewer related risk factors, but more had initiated prior chemoprophylaxis (16% vs. 0%, P < 0.01) and knew their bacille Calmette-Guerin (BCG) (68% vs. 24%, P < 0.001) and purified protein derivative (PPD) status (60% vs 32%, P < 0.05). There were no differences in symptom duration (3.3+/-3.6 vs. 3.0+/-3.4 months), mycobacteriology and diagnostic features, treatment duration (264.9+/-69.9 vs. 239.0+/-78.7 days) and completion rates (84% for both). All HCWs used self administered treatment (100% vs. 70%, P < 0.01), and fewer were hospitalized (8% vs. 28%, P < 0.05). Disease rates in nurses (3.6+/-4.4 per 100 000) were lower than the general population rates (9.0+/-0.8), but did not differ among physiotherapists (8.96+/-21.95), general practitioners (7.60+/-11.78) and medical residents (30.75+/-75.32); CONCLUSIONS: Clinical features were similar in HCWs, but management strategies differed. BC HCWs are not at increased risk of tuberculosis, but the small sample size limited the power of our study to detect such an increase. PMID- 9848611 TI - Pulmonary tuberculosis in the adult in a low prevalence area: is the radiological presentation changing? AB - SETTING: Pulmonary division of a university hospital in Belgium, an area with a relatively low tuberculosis incidence (14.9 per 100 000 population in 1993). OBJECTIVE: To evaluate whether 'unusual' radiographic presentations of pulmonary tuberculosis (PTB) have increased in recent years, especially in elderly and non indigenous patients. DESIGN: Retrospective study of chest X-rays of 219 patients aged over 18 years with bacteriologically proven PTB in the period 1981-1990, including 92 elderly (> or =65 years) and 17 non-indigenous patients. 'Unusual' presentations consisted of solitary pleural effusion, isolated hilar or mediastinal lymphadenopathies, normal chest X-ray, lower lung field TB, nodular lesions, diffuse infiltrations and atelectasis. RESULTS: There was an increase in 'unusual' presentations, from 24% in 1981-1985 to 35% in 1986-1990, yet this was not statistically significant (P = 0.08). This increase tended to be more pronounced in the elderly age group, i.e., from 24% to 42% compared to 23% to 31% in the younger age group. The proportion of elderly patients in the group with 'unusual' findings was 44% in 1981-1985 and 48% in 1986-1990. In the non indigenous group, the percentage of 'unusual' findings did not change between the two periods (40% versus 41%). The proportion of non-indigenous patients in the group with 'unusual' findings increased from 7% in 1981-1985 to 13% in 1986-1990; however, this difference was not significant. CONCLUSIONS: In an area with low TB incidence, adult PTB still presents with the classical pattern of reactivation disease. Although not statistically significant, there is a trend towards a relative increase in the frequency of unusual presentations in recent years, which is most pronounced in the elderly and in non-indigenous adults. PMID- 9848612 TI - Prevalence of pulmonary tuberculosis on the roof of the world. AB - SETTING: The Shimshal Valley, a remote village in Northern Pakistan, is one of the seven Pamirs of Central Asia, widely known as the roof of the world. OBJECTIVE: To investigate the prevalence of pulmonary tuberculosis (TB) in the Shimshal Valley. DESIGN: The Rapid Village Survey Method (RVS) was used to investigate the prevalence of pulmonary tuberculosis. The selection criteria were chronic cough, hemoptysis, past history of TB and close contact with a tuberculous patient. After clinical examination, a chest radiograph was done and a single spot sputum sample was obtained for smear examination. RESULTS: The total population of the village was 1077, of whom 231 cases were studied. Overcrowding affected 75% of the study population. The prevalence of smear positive pulmonary TB in the village studied was 554 per 100000 population, and the prevalence of active smear-negative TB was estimated at 1949/100 000. The prevalence of active pulmonary TB increased with age and the only risk factor for active TB was age over 45 years. Of the 21 cases with a past history of pulmonary TB, only 38% had completed a full course of chemotherapy. CONCLUSION: Pulmonary TB is a very serious health issue in the rural community (Shimshal Valley) of Pakistan. This study highlights the lack of efficacy of national tuberculosis control programs in the country. PMID- 9848613 TI - District sputum smear microscopy services in Malawi. AB - SETTING: Government hospitals and health centres in 23 districts in Malawi. OBJECTIVE: To determine 1) the number and smear-positivity rate of sputum samples submitted at health centres and hospitals, and 2) the time for sputum samples to get from health centres to smear examination. DESIGN: Prospective data collection on sputum specimens coming from health centres to hospital laboratories, and over the equivalent time period, retrospective data collection from laboratory sputum registers. RESULTS: Information was collected over a period of 5.6 months during 1997. Of 21 527 patients submitting sputum samples, 16995 (79%) were from within the hospital and 4532 (21%) were from health centres. Of 15 833 new TB suspects, 12 804 (81%) submitted sputum within the hospital and 3029 (19%) were from health centres. The overall smear-positivity rate was 11.9%: the proportion of new suspects who were smear-positive was significantly higher in health centres (14.1%) compared with hospital-based patients (11.4%, P < 0.05); 27% of all sputum specimens from health centres took 8 days or longer to get to smear examination. Sputum smears were positive from 1-30 days between submission and laboratory examination. CONCLUSION: Fewer sputum samples are submitted at health centres compared with hospitals, and there may be long delays between sputum submission and smear examination. The precise reasons are unclear, but health centre staff need training about the importance of timely case finding procedures. PMID- 9848614 TI - Impact of the human immunodeficiency virus epidemic on mortality among adults with tuberculosis in rural South Africa, 1991-1995. AB - SETTING: In the Hlabisa district tuberculosis programme, South Africa, the prevalence of human immunodeficiency virus (HIV) infection among adults with tuberculosis increased from 36.0% in 1993 to 65.9% in 1997, and the annual tuberculosis caseload increased from 321 in 1991 to 1250 in 1996. OBJECTIVE: To examine the impact of the HIV epidemic on mortality among adults with tuberculosis from 1991 to 1995. METHODS: Data were extracted from the control programme database. As the programme started in July 1991, data for this year were combined with those for 1992. RESULTS: All-cause mortality among all those diagnosed with tuberculosis increased by 45.6%, from 9.2% (55/599) in 1991/92 to 13.4% (96/714) in 1995 (P = 0.02). Among smear-positive patients only, mortality increased by 134%, from 4.4% to 10.3% (P = 0.003). The case-fatality rate (CFR) increased in most age groups of both sexes, the largest increase (157.5%) being among women aged 15-34 years. CFR was highest among those with smear-negative disease (24.7% in 1995). In multifactorial analysis, independent risk factors for mortality were increasing age (P = 0.0001), HIV infection (odds ratio [OR] 3.5, 95 % confidence interval [CI] 2.3-5.4), smear-negative disease (OR 2.5, 95%CI 1.8 3.5), and diagnosis in 1995 vs 1991/92 (OR 1.8, 95%CI 1.2-2.7). Mortality among the HIV infected continued to accrue throughout treatment, whereas non-HIV infected patients that died did so early in treatment. CONCLUSION: The HIV epidemic is increasing tuberculosis all-cause mortality. As well as having a direct effect on individuals, increased caseloads due to HIV may also contribute to increased mortality by reducing the health system's ability to provide adequate care. PMID- 9848615 TI - Spectrum of disease among HIV-infected adults hospitalised in a respiratory medicine unit in Abidjan, Cote d'Ivoire. AB - SETTING: Respiratory medicine wards of the University Teaching Hospital, Abidjan, Cote d'Ivoire. OBJECTIVES: To describe the spectrum of opportunistic infection among human immunodeficiency virus (HIV) infected adults hospitalised in the respiratory medicine unit in Abidjan, and the level of immunosuppression at which these diseases occur. DESIGN: Cross-sectional study. RESULTS: Overall, 75% of patients were HIV-positive: among these patients, the most frequent diagnosis was tuberculosis, in 61%, followed by bacterial pneumonia (15%), Gram-negative septicaemia (particularly non-typhoid Salmonella) (9%) and empyema (5%). Atypical pneumonias appeared to be rare. Most HIV-positive patients had CD4 counts indicative of advanced immunosuppression: 36% had CD4 counts below 100 x 10(6)/l, 19% between 100 and 199 x 10(6)/l, 29% between 200 and 499 x 10(6)/l, and 16% above 500 x 10(6)/l. Overall in-hospital mortality was 27% for HIV-positive patients and 22% for HIV-negative patients (P = 0.5). In a multivariate analysis, the strongest independent risk factors for death were cachexia (odds ratio [OR] 7.4, 95% confidence interval [CI] 2.1-26.3), male sex (OR 4.5, 95% CI 1.2-17.4) and age over 40 (OR 4.1, 95% CI 1.0-17.2). CONCLUSIONS: Tuberculosis and bacterial infections are the major causes of respiratory morbidity in immunosuppressed HIV-infected adults in this population. Efforts to improve the management of HIV-related disease need to focus on prevention and treatment of these infections. PMID- 9848616 TI - Evaluation of Roche Amplicor PCR assay for Mycobacterium avium complex in bronchial washing. AB - SETTING: A commercially available polymerase chain reaction (PCR) test (Roche AmplicorTM Mycobacterium avium and M. intracellulare assay-MAC-PCR) designed to detect M. avium complex (MAC) in bronchial washing was evaluated. DESIGN: A total of 141 specimens from 127 patients with various pulmonary conditions were examined. Results were compared with acid-fast smears, cultures with Ogawa egg medium, as is still commonly used in Japan, and final diagnoses. RESULTS AND CONCLUSIONS: A total of 14 bronchial washing specimens yielded MAC. Six smear- and culture-positive specimens were all MAC-PCR positive. In eight smear-negative and culture-positive specimens, six were MAC-PCR positive. The overall sensitivity versus culture was 85.7% (12/14). However, sensitivity might be over estimated, as there is a lower recovery rate of MAC with egg-based medium compared with liquid media. In 127 patients, 15 were identified as having pulmonary MAC disease, of whom 13 had positive MAC-PCR in bronchial washing. In the remaining 112 patients, MAC-PCR was negative, which suggests that positive MAC-PCR was not a contaminated result. However, in terms of sensitivity and speed, we were unable to show any additional clinical benefit for using MAC-PCR as opposed to liquid media, in which MAC can frequently be detected in 7 to 14 days. PMID- 9848617 TI - Tuberculous pleural effusions in HIV-positive patients. PMID- 9848618 TI - Severe arthropathy with ofloxacin in two cases of MDR tuberculosis. PMID- 9848619 TI - DOTS and beyond: towards a holistic approach to the conquest of tuberculosis. PMID- 9848620 TI - Registration and treatment of patients with smear-positive pulmonary tuberculosis. PMID- 9848621 TI - Tuberculin PPD RT23 is stable: quality control data cannot be denied. PMID- 9848622 TI - Human health effects of air pollution from mobile sources in Europe. PMID- 9848623 TI - Calculation of the protein equivalent of total nitrogen appearance from urea appearance. Which formulas should be used? PMID- 9848624 TI - Should dialysate calcium be varied in proportion to the amount of ultrafiltration in peritoneal dialysis dwells? Directions from a computer simulation. PMID- 9848625 TI - Comparing mortality rates on CAPD/CCPD and hemodialysis. The Canadian experience: fact or fiction? AB - OBJECTIVE: To compare mortality rates on hemodialysis (HD) to rates on continuous ambulatory/cyclic peritoneal dialysis (CAPD/CCPD), to contrast our results with those of other recent investigations, and to discuss reasons for discrepancies. DATA SOURCES: Patient-specific data obtained from the Canadian Organ Replacement Register on patients initiating renal replacement therapy (RRT) between 1 January 1990 and 31 December 1995 (n = 14 483). Recent mortality comparisons of CAPD and HD. MAIN OUTCOME MEASURES: Mortality rate ratio (RR) based on "as-treated" (AT) analysis incorporating treatment modality switches and adjusting for age, primary renal diagnosis, and comorbid conditions using Poisson regression. Hazard ratios (HR) were estimated using Cox regression and based on an "intent-to-treat" (ITT) analysis wherein patients were classified based on dialytic modality received on follow-up day 90. RESULTS: Adjusted mortality rates were significantly decreased on CAPD/CCPD relative to HD [RR = 0.73, 95% confidence interval (CI) = (0.69, 0.77)] based on the AT analysis. Most of the protective effect of CAPD/CCPD was concentrated in the first 2 years of follow-up post-RRT initiation. Based on the ITT analysis, the estimated CAPD/ CCPD effect was greatly reduced, with HR = 0.93 (0.87, 0.99). CONCLUSIONS: We provide further evidence that CAPD/CCPD is not an inferior dialytic modality to HD, particularly in the short term. Comparing mortality rates on CAPD/CCPD and HD is inherently difficult due to the potential for bias. Discrepancies between our results and those of previous investigations, and variability in findings among previous studies, relate to differences in clinical and demographic setting, patient populations, study design, statistical methods, and interaction between the dialytic modality effect and various other covariables. PMID- 9848626 TI - The futility of pretransplant coronary bypass grafting in asymptomatic patients on peritoneal dialysis. AB - OBJECTIVE: To evaluate the validity of recommending coronary artery bypass grafting (CABG) in preparation for renal transplantation in asymptomatic peritoneal dialysis (PD) patients with evidence of reversible myocardial ischemia. DESIGN: Retrospective review in a single PD unit. PARTICIPANTS: Ten asymptomatic PD patients who underwent CABG to be placed on the transplant list comprised the study group. Ten age-, sex-, and disease-matched PD patients who did not receive CABG were used as a comparison group. MEASUREMENTS: Clinical outcome from 1990 to the present. RESULTS: Only 1 patient in the study group has received a transplant. Seven patients (70%) have died or have been removed from the list because of comorbid illness. Only 2 patients are still on the waiting list. CONCLUSION: As a result of the long waiting time for cadaveric renal transplant and the high risk of interim development of comorbid disease, only a minority of patients come to transplantation. The presence of coronary disease is likely a surrogate for more generalized cardiac and vascular disease in this population. In light of these findings, the policy of prophylactic revascularization in asymptomatic dialysis patients in preparation for renal transplantation needs to be reconsidered. PMID- 9848627 TI - 1996 Peritoneal Dialysis Core Indicators Study: report on nutritional indicators. AB - OBJECTIVE: The 1996 Peritoneal Dialysis Core Indicators Study illustrates the conduct of peritoneal dialysis in the United States during 1996. DESIGN AND PATIENT POPULATION: The survey is a medical records audit of 1317 randomly selected adult U.S.A. Medicare patients using peritoneal dialysis during 1996. OUTCOME MEASURES: Abstracted data included basic demographic characteristics, dialysis prescription, delivered dialysis dose, residual renal function, serum albumin, hematocrit, anemia management, and patient status. RESULTS: The survey included 785 patients using continuous ambulatory peritoneal dialysis (CAPD) and 423 using automated peritoneal dialysis (APD) primarily in the form of continuous cycling peritoneal dialysis (CCPD). Except for the prescription mechanics and a greater likelihood that African-Americans would use CAPD, the groups did not differ substantially from one another. Evaluation of patient weight (W), body mass index (BMI), residual renal function, average serum albumin, protein equivalent of nitrogen appearance (nPNA), and dialysis efficiency as weekly fractional urea nitrogen removal (wKt/Vurea) and weekly creatinine clearance (wCrCl) revealed a picture of reasonable dialysis delivery and marginal protein nutrition. Additionally, there was little evidence that "dialysis efficiency," over the range assessed, had a major influence on nutritional status. Despite a tendency toward obesity (body weight = 76.6+/-20.0 kg and BMI = 27+/-7), 47% of patients had an average serum albumin below"normal" (3.5 g/dL by bromcresol green) and 70% had a nPNA below 1.0 g/kg/day. CONCLUSIONS: Peritoneal dialysis patients appear to have marginal protein reserves despite surfeit energy stores. PMID- 9848628 TI - Clearance of tracer albumin from peritoneal cavity to plasma at low intraperitoneal volumes and hydrostatic pressures. AB - OBJECTIVE: To assess the clearance of radiolabeled tracer albumin (RISA) from peritoneal cavity to plasma (Cl-->P) in rats under essentially "normal" conditions, that is, when intraperitoneal hydrostatic pressure (IPP) is subatmospheric and the intraperitoneal (IP) "free" fluid volume (IPV) is low. METHODS: A volume of 0.3 mL of RISA was injected IP into anesthetized Wistar rats (wt = 300 g) when the IPV was approximately 2 mL (normal) or the IPV was approximately 10 mL, and IPP was either -1.8 mmHg (normal) or +1.5 mmHg (produced by an external cuff). Plasma samples (25 microL) were obtained repeatedly during the dwell, which lasted 30-300 min, after which the peritoneal cavity was opened to recover the IPV and residual IP RISA activity. The Cl-->P was assessed as the mass transfer of RISA into plasma, occurring per unit time, divided by the calculated mean IP RISA concentration (CD). The interstitial RISA space was measured as the mass of RISA accumulated, per unit tissue weight, in peritoneal tissue samples divided by the CD. RESULTS: A markedly lower Cl-->P (2.47+/-0.67 microL/min), as well as total RISA clearance out of the peritoneal cavity (Cl), was found under "normal" conditions (an IPV of approximately 2 mL and an IPP of approximately -1.8 mmHg) compared to the situation during peritoneal dialysis (an IPV of approximately 20 mL and an IPP of +1 mmHg). Furthermore, the interstitial RISA space increased linearly over time even at negative IPPs and at an unchanging peritoneal interstitial fluid volume. At a low (normal) IPV the Cl-->P did not increase significantly with elevating IPP, and increased only marginally when tracer distribution was improved by artificial vibration of the rats. However the Cl-->P increased when larger volumes were infused to increase the total IPV. CONCLUSIONS: It is concluded that the Cl-->P and Cl at low IPPs and IPVs are not as high as during peritoneal dialysis. Increases in Cl-->P were, however, coupled to increases in IPV. This highlights the importance of the IPV per se and of a sufficient IP tracer distribution for direct lymphatic absorption to be efficient. PMID- 9848630 TI - Plasma iohexol clearance in automated peritoneal dialysis--its role in adequacy determination. AB - OBJECTIVE: To compare the relationship of plasma iohexol clearance to standard adequacy measures in an automated peritoneal dialysis (APD) population. DESIGN: Prospective, nonrandomized, open label, simultaneous clearance studies of novel (iohexol) and traditional (urea and creatinine) markers of dialysis quantitation. SETTING: Outpatient peritoneal dialysis units associated with tertiary care university hospitals. PATIENTS: Eighteen stable patients, 13 undergoing continuous cycling peritoneal dialysis (CCPD) and 5 receiving intermittent dialysis, who underwent 24-hour clearance studies were enrolled in and completed the study. INTERVENTIONS: Each subject received 15 mL of iohexol intravenously the morning of a scheduled dialysis night. Blood was obtained for determination of serum concentrations of urea nitrogen and creatinine, and plasma iohexol concentration. Dialysate and urine were collected over 24 hours. MAIN OUTCOME MEASURES: Urea and creatinine data from the total pooled dialysate and from the 24-hour urine collection were used to calculate daily Kt/Vurea and normalized total daily creatinine clearance (CrCl). Total plasma iohexol clearance was calculated using a one-compartment model with the Brochner-Mortensen correction. RESULTS: Normalized [to 1.73 m2 body surface area (BSA)] iohexol clearance correlated very well with normalized CrCl (r2 = 0.777; p < 0.001). A weaker correlation was observed between Kt/Vurea and normalized iohexol clearance (r2 = 0.213; 0.05 < p < 0.1). When data are not normalized using BSA or urea distribution volume, the regressions are improved for both creatinine (r2 = 0.820) and urea (r2 = 0.533). These results were comparable between those on CCPD and those on intermittent therapy. CONCLUSION: Total plasma iohexol clearance provides a simple assessment for APD adequacy by eliminating collection problems inherent to the methodologies currently employed. Such simplicity allows for more frequent assessments of delivered dialysis dose and efficacy of prescription changes. PMID- 9848629 TI - Effects of oral glucose on intermediary metabolism in continuous ambulatory peritoneal dialysis patients versus healthy subjects. AB - OBJECTIVE: To explore the mechanisms and metabolic consequences of the insulin resistance of patients treated by continuous ambulatory peritoneal dialysis (CAPD). DESIGN: CAPD patients and healthy subjects ingested a similar mean oral glucose load per kilogram of fat-free mass (FFM) [1.20+/-0.03 g/(kg FFM) vs 1.20+/-0.06 g/(kg FFM); CAPD vs healthy subjects]. Substrate oxidation was monitored over 6 hours using indirect calorimetry. SETTING: Peritoneal dialysis unit of a tertiary-care institutional center. OUTCOME MEASURES: Glycemia, insulinemia, substrate oxidation. PATIENTS: Six CAPD patients (68+/-5 yr) and 6 healthy subjects (24+/-1 yr). The CAPD patients had similar body mass index (21.4+/-1.3 vs 22.9+/-1.1 kg/m2), a higher percent fat (25.8%+/-3.7% vs 16%+/ 2.2%; p < 0.05), and a lower FFM (42.2+/-2.2 kg vs 56.5+/-2.6 kg; p < 0.01) than healthy subjects. RESULTS: The CAPD patients displayed a higher glycemic and insulinemic responses to glucose than did healthy subjects (p < 0.05), but similar glucose oxidation and storage. Lipid oxidation and plasma nonesterified fatty acids were not increased in CAPD patients versus healthy subjects, in spite of a higher adiposity. Fat oxidation was related to fat mass in CAPD patients (r2 = 0.77, p < 0.05) but not in healthy subjects (r2 = 0.05). CONCLUSION: CAPD patients display an insulin-resistance not explained by an increased lipid oxidation. The maintenance of intracellular glucose utilization at the expense of higher glycemic and insulinemic responses suggests a defective glucose transport. PMID- 9848631 TI - Thyroid dysfunction and nodular goiter in hemodialysis and peritoneal dialysis patients. AB - OBJECTIVE: To investigate the prevalence of nodular goiter and thyroid dysfunction in uremic patients undergoing hemodialysis (HD) and peritoneal dialysis. DESIGN: Cross-sectional study. SETTING: Single dialysis unit and outpatient clinic. PATIENTS: The study included 221 patients [143 HD and 78 continuous ambulatory peritoneal dialysis (CAPD) patients] along with 135 consecutively selected outpatients as controls. MAIN OUTCOME MEASURES: Ultrasonography was used to detect patients' thyroid function and nodular goiter. RESULTS: Nodular goiter was detected in 54.8% of the uremic patients and in 21.5% of the controls. Uremic patients had higher prevalence of thyroid dysfunction, which included reduced serum concentration of total T3, total T4, and free T4, and increased serum level of TSH. Hypothyroidism was also observed more frequently in uremic patients than in the control group (5.4% vs 0.7%, p < 0.05). Nodular goiter was more frequently found in females than in males (63.5% vs 48%, p < 0.05). Moreover, the prevalence of nodular goiter increased with age (p < 0.02) in uremic patients. Hemodialysis patients had a higher frequency of reduced total T3 level (46.9% vs 29.5%, p < 0.02). However, CAPD patients had lower T4 levels (6.23+/-1.82 microg/dL vs 7.15+/-1.99 microg/dL, p < 0.05). CONCLUSION: Because of the high incidence of hypothyroidism and nodular goiter in uremic patients, screening of thyroid function and goiter detection with ultrasound should be considered in evaluation of end-stage renal disease patients. PMID- 9848632 TI - Safety and efficacy of total dose iron dextran administration in patients on home renal replacement therapies. AB - OBJECTIVE: To determine the safety and efficacy of intravenous total dose iron (TDI) replacement in patients treated with home renal replacement therapy. DESIGN: Prospective open-label study on end points in the population studied. SETTING: Institutional outpatient home dialysis program. PATIENTS: The study included 20 end-stage renal disease (ESRD) patients, performing chronic peritoneal or home hemodialysis, with iron deficiency defined as ferritin < 100 ng/mL and/or an iron saturation < 20%. INTERVENTION: The total dose of iron dextran was calculated and infused at a rate not exceeding 6 mg/min. Hemoglobin, hematocrit, iron studies, and liver function tests (LFTs) were obtained before and 3 to 4 weeks after TDI infusion. Hematocrit of patients failing to achieve an increase in Hct over this period was re-examined 2 to 4 weeks later looking for a delayed response. MAIN OUTCOME MEASURES: Primary end points for efficacy were changes in Hct, ferritin, and iron saturation. Toxicity was measured as reported immediate and delayed symptoms and elevated transaminases and/or alkaline phosphatase levels. RESULTS: A median iron dose of 1000 mg (range, 325-1500 mg) was administered. The infusions were generally well tolerated. Clinical adverse effects were seen in 2 patients weighing less than 50 kg. No increase in LFT results was seen. Hematocrit increased 2.2% (95% CI, 0.5%-3.9%) from 29.0% to 31.2% (p = 0.01) within 4 weeks of infusion. Significant increases also occurred in iron saturation (from 13% to 22%, p = 0.001) and ferritin (from 234 to 305 ng/mL, p = 0.008). Among the 9 patients who did not respond with a significant increase in Hct, 2 had a delayed response, increasing the overall response from 63% at 4 weeks to 71%, 8 weeks after TDI. Inadequate erythropoietin dosing and low-grade infectious/inflammatory disorders may have contributed to a poor response in several patients. CONCLUSION: Total dose iron is a safe and effective means of restoring iron and erythropoietic response in ESRD patients weighing more than 50 kg who receive their renal replacement therapy at home. PMID- 9848633 TI - A survey of home visits at peritoneal dialysis centers in the United States. AB - OBJECTIVE: To determine the frequency and characteristics of home visits in centers that provide training for peritoneal dialysis (PD). DESIGN: Mail survey sent to all dialysis centers in the United States providing home PD, using the Health Care Federal Administration (HCFA) Renal Provider list. RESULTS: Surveys were mailed to 1247 centers; 13 were undeliverable, resulting in 1234 surveys successfully delivered; 670 (54%) of those surveyed responded. Of those responding, 525 (78.4%) reported home visits were part of the care of home PD patients: 11% made a single home visit, 52% made an initial home visit with at least one follow-up visit, and 16% made visits only as needed. No home visits were made by 21% of responding centers. A registered nurse (RN) alone made the home visit in 61% of the centers, while a multidisciplinary team accompanied the RN in 35% of centers; 3% of visits were made by a licensed practical nurse, and 1% by the physician. Half of the visits required 0.5-1 hour, while 41% required 1 2 hours. Travel time was most often an hour or less one way. Staff were reimbursed for travel expenses by 90% of the centers. The 525 centers making home visits were not different than the 145 centers not making home visits in number of patients per center, number of RNs, rural or urban location, or affiliation with a university. Interpretation of the HCFA regulations concerning home visits was the most important factor influencing centers making home visits. CONCLUSIONS: Home visits to continuous ambulatory PD and continuous cycling PD patients in the United States are common. Nearly 80% of centers responding to the survey include home visits in the care of their home peritoneal dialysis patients. PMID- 9848634 TI - The effect of octreotide on peritoneal alterations induced by high concentration of glucose. PMID- 9848635 TI - CAPD in patients with failure of hemodialysis vascular access. PMID- 9848636 TI - Diarrhea induced by CAPD. A new complication following initiation of CAPD? PMID- 9848637 TI - Peritoneal dialysis in a patient receiving mechanical ventilation in prone position. PMID- 9848638 TI - Stability of iron dextran (DexFerrum) in peritoneal dialysis bags. PMID- 9848639 TI - Staphylococcus aureus nasal carriage in pediatric peritoneal dialysis patients and their parents. PMID- 9848640 TI - Inflammatory demyelinating neuropathy presenting in a temporal relationship with the initiation of peritoneal dialysis. PMID- 9848641 TI - Successful treatment of peritonitis in CAPD with intraperitoneal rifampin. PMID- 9848642 TI - Peritonitis caused by Clostridium perfringens in a CAPD patient. PMID- 9848643 TI - Peritoneal dialysis case forum. Necrotizing panniculitis in a morbidly obese patient with end-stage renal disease. PMID- 9848644 TI - Literature. September-October 1998. PMID- 9848645 TI - Nursing application: results aid in identifying focus of patient teaching. PMID- 9848646 TI - Directed hydroxyl radical probing of 16S ribosomal RNA in ribosomes containing Fe(II) tethered to ribosomal protein S20. AB - The 16S ribosomal RNA neighborhood of ribosomal protein S20 has been mapped, in both 30S subunits and 70S ribosomes, using directed hydroxyl radical probing. Cysteine residues were introduced at amino acid positions 14, 23, 49, and 57 of S20, and used for tethering 1-(p-bromoacetamidobenzyl)-Fe(II)-EDTA. In vitro reconstitution using Fe(II)-derivatized S20, together with the remaining small subunit ribosomal proteins and 16S ribosomal RNA (rRNA), yielded functional 30S subunits. Both 30S subunits and 70S ribosomes containing Fe(II)-S20 were purified and hydroxyl radicals were generated from the tethered Fe(II). Hydroxyl radical cleavage of the 16S rRNA backbone was monitored by primer extension. Different cleavage patterns in 16S rRNA were observed from Fe(II) tethered to each of the four positions, and these patterns were not significantly different in 30S and 70S ribosomes. Cleavage sites were mapped to positions 160-200, 320, and 340-350 in the 5' domain, and to positions 1427-1430 and 1439-1458 in the distal end of the penultimate stem of 16S rRNA, placing these regions near each other in three dimensions. These results are consistent with previous footprinting data that localized S20 near these 16S rRNA elements, providing evidence that S20, like S17, is located near the bottom of the 30S subunit. PMID- 9848647 TI - In vitro selection of the Naegleria GIR1 ribozyme identifies three base changes that dramatically improve activity. AB - NanGIR1 is a member of a new class of group I ribozymes whose putative biological function is site-specific hydrolysis at an internal processing site (IPS). We have previously shown that NanGIR1 requires 1 M KCl for maximal activity, which is nevertheless slow (0.03 min(-1)). We used in vitro selection and an RNA pool with approximately nine mutations per molecule to select for faster hydrolysis at the IPS in 100 mM KCl. After eight rounds of selection, GIR1 variants were isolated that catalyzed hydrolysis at 300-fold greater rates than NanGIR1 RNA. Although not required by the selection, many of the resultant RNAs had increased thermal stability relative to the parent RNA, and had a more compact structure as evidenced by their faster migration in native gels. Although a wide spectrum of mutations was found in generation 8 clones, only two mutations, U149C and U153C, were common to greater than 95% of the molecules. These and one other mutation, G32A, are sufficient to increase activity 50-fold. All three mutations lie within or proximal to the P15 pseudoknot, a structural signature of GIR1 RNAs that was previously shown to be important for catalytic activity. Overall, our findings show that variants of the Naegleria GIR1 ribozyme with dramatically improved activity lie very close to the natural GIR1 in sequence space. Furthermore, the selection for higher activity appeared to select for increased structural stability. PMID- 9848648 TI - The snRNP-free U1A (SF-A) complex(es): identification of the largest subunit as PSF, the polypyrimidine-tract binding protein-associated splicing factor. AB - We have previously shown that a specific monoclonal antibody prepared against the U1A protein, MAb 12E12, is unique in its ability to recognize a form of U1A which is not associated with the U1snRNP. This unique form of U1A, termed snRNP-free U1A or SF-A, was found to be complexed with a novel set of non-snRNP proteins (O'Connor et al., 1997, RNA 3:1444-1455). Here we demonstrate that the largest protein in these SF-A complex(es), p105, is the polypyrimidine-tract binding protein-associated factor (PSF), an auxiliary splicing factor. We show that PSF copurifies and co-immunoprecipitates with SF-A from 293T cell nucleoplasm and that it interacts with SF-A in vitro. In addition, we show that MAb 12E12 inhibits both splicing and polyadenylation in an in vitro coupled splicing and polyadenylation reaction. This suggests that SF-A and/or the SF-A complex(es) perform an important function in both processing reactions and possibly in last exon definition. PMID- 9848649 TI - Cap-independent polysomal association of natural mRNAs encoding c-myc, BiP, and eIF4G conferred by internal ribosome entry sites. AB - Sequence elements that can function as internal ribosome entry sites (IRES) have been identified in 5' noncoding regions of certain uncapped viral and capped cellular mRNA molecules. However, it has remained largely unknown whether IRES elements are functional when located in their natural capped mRNAs. Therefore, the polysomal association and translation of several IRES-containing cellular mRNAs was tested under conditions that severely inhibited cap-dependent translation, that is, after infection with poliovirus. It was found that several known IRES-containing mRNAs, such as BiP and c-myc, were both associated with the translation apparatus and translated in infected cells when cap-dependent translation of most host-cell mRNAs was blocked, indicating that the IRES elements were functional in their natural mRNAs. Curiously, the mRNAs that encode eukaryotic initiation factor 4GI (eIF4GI) and 4GII (eIF4GII), two proteins with high identity and similar functions in the initiation of cap-dependent translation, were both associated with polysomes in infected cells. The 5'-end sequences of eIF4GI mRNA were isolated from a cDNA expression library and shown to function as an internal ribosome entry site when placed into a dicistronic mRNA. These findings suggest that eIF4G proteins can be synthesized at times when 5' cap-dependent mRNA translation is blocked, supporting the notion that eIF4G proteins are needed in both 5' cap-independent and 5' cap-dependent translational initiation mechanisms. PMID- 9848650 TI - tRNA(2Gln) mutants that translate the CGA arginine codon as glutamine in Escherichia coli. AB - We present a novel missense suppression system for the selection of tRNA(2GIn) mutants that can efficiently translate the CGA (arginine) codon as glutamine. tRNA(2Gln) mutants were cloned from a partially randomized synthetic gene pool using a plasmid vector that simultaneously expresses the tRNA gene and, to ensure efficient aminoacylation, the glutamine aminoacyl-tRNA synthetase gene (glnS). tRNA mutants that insert glutamine at CGA were selected as missense suppressors of a lacZ mutant (lacZ625(CGA)) that contains CGA substituted for an essential glutamine codon. Preliminary characterizations of four suppressors is presented. All of them contain two anticodon mutations: C-->U at position 34 and U-->C at position 35, which allow for cognate translation of CGA. U35 was previously shown to be an important determinant for glutaminylation of tRNA(2Gln) in vitro; suppression in vivo requires overexpression of the glutaminyl-tRNA synthetase gene (glnS). One tRNA variant contains no further mutations and has the highest missense suppression activity (8%). Three other isolates each contain an additional point mutation that alters suppression efficiency. This system will be useful for further studies of tRNA structure and function. In addition, because relatively efficient translation of the rare CGA codon as glutamine is not toxic for Escherichia coli, it may be possible to translate this sense codon with other alternate meanings, a property which could greatly facilitate protein engineering. PMID- 9848651 TI - Enhancer elements activate the weak 3' splice site of alpha-tropomyosin exon 2. AB - We have identified four purine-rich sequences that act as splicing enhancer elements to activate the weak 3' splice site of alpha-tropomyosin exon 2. These elements also activate the splicing of heterologous substrates containing weak 3' splice sites or mutated 5' splice sites. However, they are unique in that they can activate splicing whether they are placed in an upstream or downstream exon, and the two central elements can function regardless of their position relative to one another. The presence of excess RNAs containing these enhancers could effectively inhibit in vitro pre-mRNA splicing reactions in a substrate-dependent manner and, at lower concentrations of competitor RNA, the addition of SR proteins could relieve the inhibition. However, when extracts were depleted by incubation with biotinylated exon 2 RNAs followed by passage over streptavidin agarose, SR proteins were not sufficient to restore splicing. Instead, both SR proteins and fractions containing a 110-kD protein were necessary to rescue splicing. Using gel mobility shift assays, we show that formation of stable enhancer-specific complexes on alpha-tropomyosin exon 2 requires the presence of both SR proteins and the 110-kD protein. By analogy to the doublesex exon enhancer elements in Drosophila, our results suggest that assembly of mammalian exon enhancer complexes requires both SR and non-SR proteins to activate selection of weak splice sites. PMID- 9848652 TI - A polysomal ribonuclease involved in the destabilization of albumin mRNA is a novel member of the peroxidase gene family. AB - We have purified an approximately 60 kDa endoribonuclease from Xenopus liver polysomes with properties expected for a messenger RNase involved in the estrogen regulated destabilization of serum protein mRNAs (Dompenciel et al., 1995, J Biol Chem 270:6108-6118). The present report describes the cloning of this protein and its identification as a novel member of the peroxidase gene family. This novel enzyme, named polysomal RNase 1, or PMR-1 has 57% sequence identity with myeloperoxidase, and like that protein, appears to be processed from a larger precursor. Unlike myeloperoxidase, however, PMR-1 lacks N-linked oligosaccharide, heme, and peroxidase activity. Western blot and immunoprecipitation experiments using epitope-specific antibodies to the derived protein sequence confirm the identity of the cloned cDNA to the protein originally isolated from polysomes. The 80 kDa pre-PMR-1 expressed in a recombinant baculovirus was not processed to the 60 kDa form in Sf9 cells and lacks RNase activity. However, the baculovirus expressed mature 60-kDa form of the enzyme has RNase activity. The recombinant protein is an endonuclease that shows selectivity for albumin versus ferritin mRNA. While it does not cleave at consensus APyrUGA elements, recombinant PMR-1 generates the same minor cleavage products from albumin mRNA as PMR-1 purified from liver. Finally, we show estrogen induces only a small increase in the amount of PMR-1. This result is consistent with earlier data suggesting estrogen activates mRNA decay through a posttranslational pathway. PMID- 9848653 TI - Cbf5p, a potential pseudouridine synthase, and Nhp2p, a putative RNA-binding protein, are present together with Gar1p in all H BOX/ACA-motif snoRNPs and constitute a common bipartite structure. AB - The eukaryotic nucleolus contains a large number of small nucleolar RNAs (snoRNAs) that are involved in preribosomal RNA (pre-rRNA) processing. The H box/ACA-motif (H/ACA) class of snoRNAs has recently been demonstrated to function as guide RNAs targeting specific uridines in the pre-rRNA for pseudouridine (psi) synthesis. To characterize the protein components of this class of snoRNPs, we have purified the snR42 and snR30 snoRNP complexes by anti-m3G-immunoaffinity and Mono-Q chromatography of Saccharomyces cerevisiae extracts. Sequence analysis of the individual polypeptides demonstrated that the three proteins Gar1p, Nhp2p, and Cbf5p are common to both the snR30 and snR42 complexes. Nhp2p is a highly basic protein that belongs to a family of putative RNA-binding proteins. Cbf5p has recently been demonstrated to be involved in ribosome biogenesis and also shows striking homology with known prokaryotic psi synthases. The presence of Cbf5p, a putative psi synthase in each H/ACA snoRNP suggests that this class of RNPs functions as individual modification enzymes. Immunoprecipitation studies using either anti-Cbf5p antibodies or a hemagglutinin-tagged Nhp2p demonstrated that both proteins are associated with all H/ACA-motif snoRNPs. In vivo depletion of Nhp2p results in a reduction in the steady-state levels of all H/ACA snoRNAs. Electron microscopy of purified snR42 and snR30 particles revealed that these two snoRNPs possess a similar bipartite structure that we propose to be a major structural determining principle for all H/ACA snoRNPs. PMID- 9848654 TI - The rhinovirus type 14 genome contains an internally located RNA structure that is required for viral replication. AB - Cis-acting RNA signals are required for replication of positive-strand viruses such as the picornaviruses. Although these generally have been mapped to the 5' and/or 3' termini of the viral genome, RNAs derived from human rhinovirus type 14 are unable to replicate unless they contain an internal cis-acting replication element (cre) located within the genome segment encoding the capsid proteins. Here, we show that the essential cre sequence is 83-96 nt in length and located between nt 2318-2413 of the genome. Using dicistronic RNAs in which translation of the P1 and P2-P3 segments of the polyprotein were functionally dissociated, we further demonstrate that translation of the cre sequence is not required for RNA replication. Thus, although it is located within a protein-coding segment of the genome, the cre functions as an RNA entity. Computer folds suggested that cre sequences could form a stable structure in either positive- or minus-strand RNA. However, an analysis of mutant RNAs containing multiple covariant and non covariant nucleotide substitutions within these putative structures demonstrated that only the predicted positive-strand structure is essential for efficient RNA replication. The absence of detectable minus-strand synthesis from RNAs that lack the cre suggests that the cre is required for initiation of minus-strand RNA synthesis. Since a lethal 3' noncoding region mutation could be partially rescued by a compensating mutation within the cre, the cre appears to participate in a long-range RNA-RNA interaction required for this process. These data provide novel insight into the mechanisms of replication of a positive-strand RNA virus, as they define the involvement of an internally located RNA structure in the recognition of viral RNA by the viral replicase complex. Since internally located RNA replication signals have been shown to exist in several other positive-strand RNA virus families, these observations are potentially relevant to a wide array of related viruses. PMID- 9848656 TI - Trans-splicing group II introns in plant mitochondria: the complete set of cis arranged homologs in ferns, fern allies, and a hornwort. AB - The fragmentation of group II introns without concomitant loss of splicing competence is illustrated by extraordinary gene arrangements in plant mitochondrial genomes. The mitochondrial genes nad1, nad2, and nad5, all encoding subunits of the NADH dehydrogenase, require trans-splicing for functional assembly of their mRNAs in flowering plants. Tracing the origins of trans splicing group II introns shows that they have evolved from formerly cis-arranged homologs whose descendants can still be identified in lineages of early branching land plants. In this contribution we present the full set of ancestor introns for all five conserved mitochondrial trans-splicing positions. These introns are strikingly small in the quillwort Isoetes lacustris, the continuous nad2 gene intron in this species representing the smallest (389 nt) land plant group II intron yet identified. cDNA analysis shows correct splicing of the introns in vivo and also identifies frequent RNA editing events in the flanking nad gene exons. Other representatives of the ancestral cis-arranged introns are identified in the fern Osmunda regalis, the horsetail Equisetum telmateia, and the hornwort Anthoceros crispulus. Only the now identified intron in Osmunda carries significant traces of a former maturase reading frame. The identification of a continuous homolog in Anthoceros demonstrates that intron invasion into the affected genes in some cases predated the split of vascular and nonvascular plants more than 400 million years ago. As an alternative to disruption after size increase, the respective introns can get secondarily lost in certain lineages. PMID- 9848655 TI - Separable roles in vivo for the two RNA binding domains of Drosophila A1-hnRNP homolog. AB - We analyzed the roles of the three domains of a Drosophila hnRNP A1 homolog by expression of wild-type and mutant versions of HRB87F/hrp36 in Drosophila melanogaster. HRB87F/hrp36 is one of two Drosophila proteins that is most similar to mammalian A1 hnRNP, and like A1, consists of two copies of the RNA-binding domain (RBD) motif followed by a glycine-rich domain (GRD). The role of the domains in nuclear localization and RNA binding to polytene chromosomal sites was determined. RBD-1 and the GRD were largely responsible for both the cellular location of the protein and for the typical chromosomal distribution pattern of the protein at sites of PolII transcription. RBD-1 also provided a role in the exon-skipping activity of the protein that was not provided by RBD-2. On the other hand, RBD-2 and the GRD were responsible for the very limited chromosomal distribution pattern seen upon heat shock, when HRB87F/hrp36 is sequestered at heat-shock puff 93D, which encodes a long nucleus-restricted RNA. Thus, these studies indicate that the two RBDs function independently of each other but in concert with the GRD. In addition, the self-association property of the GRD was strikingly evident in these overexpressed proteins. PMID- 9848657 TI - The structure of the ITS2-proximal stem is required for pre-rRNA processing in yeast. AB - Accurate and efficient processing of pre-rRNA is critical to the accumulation of mature functional ribosomal subunits for maintenance of cell growth. Processing requires numerous factors which act in trans as well as RNA sequence/ structural elements which function in cis. To examine the latter, we have used directed mutagenesis and expression of mutated pre-rRNAs in yeast. Specifically, we tested requirements for formation of an ITS2-proximal stem on processing, a structure formed by an interaction between sequences corresponding to the 3' end of 5.8S rRNA and the 5' end of 25S. Pre-rRNA processing is inhibited in templates encoding mutations that prevent the formation of the ITS2-proximal stem. Compensatory, double mutations, which alter the sequence of this region but restore the structure of the stem, also restore processing, although at lower efficiency. This reduction in efficiency is reflected in decreased levels of mature 5.8S and 25S rRNA and increased levels of 35S pre-rRNA and certain processing intermediates. This phenotype is reminiscent of the biochemical depletion of U8 snoRNA in vertebrates for which the ITS2-proximal stem has been proposed as a potential site for interaction with U8 RNP. Thus, formation of the ITS2-proximal stem may be a requirement common to yeast and vertebrate pre-rRNA processing. PMID- 9848658 TI - Identification of RNA sequences and structures involved in site-specific cleavage of IGF-II mRNAs. AB - Insulin-like growth factor-II (IGF-II) mRNAs are subject to site-specific endonucleolytic cleavage in the 3' untranslated region (UTR), rendering an unstable 5' cleavage product containing the coding region and a very stable 3' cleavage product of 1.8 kb consisting of the 3'-UTR sequence and the poly(A) tail. Previously, it was established that two widely separated elements in the 3' UTR (elements I and II), that can form a duplex structure, are necessary and sufficient for cleavage. To further investigate the sequence and secondary structure requirements for cleavage, we have introduced a number of mutations around the cleavage site and assayed their effects on cleavage. Several recognition determinants involved in the endonucleolytic cleavage of IGF-II mRNAs were identified. Mutational analysis around the cleavage site revealed that cleavage is sequence specific and that the cleavage site must be in a single stranded conformation to allow efficient cleavage. In addition, we have identified an accessory protein that specifically interacts with a stem-loop structure located 133 to 73 nt upstream of the cleavage site. PMID- 9848659 TI - Two mutant forms of the S1/TPR-containing protein Rrp5p affect the 18S rRNA synthesis in Saccharomyces cerevisiae. AB - The genetic depletion of yeast Rrp5p results in a synthesis defect of both 18S and 5.8S ribosomal RNAs (Venema J, Tollervey D. 1996. EMBO J 15:5701-5714). We have isolated the RRP5gene in a genetic approach aimed to select for yeast factors interfering with protein import into mitochondria. We describe here a striking feature of Rrp5p amino acid sequence, namely the presence of twelve putative S1 RNA-binding motifs and seven tetratricopeptide repeats (TPR) motifs. We have constructed two conditional temperature-sensitive alleles of RRP5 gene and analyzed them for associated rRNA-processing defects. First, a functional "bipartite gene" was generated revealing that the S1 and TPR parts of the protein can act independently of each other. We also generated a two amino acid deletion in TPR unit 1 (rrp5delta6 allele). The two mutant forms of Rrp5p were shown to cause a defect in 18S rRNA synthesis with no detectable effects on 5.8S rRNA production. However, the rRNA processing pathway was differently affected in each case. Interestingly, the ROK1 gene which, like RRP5, was previously isolated in a screen for synthetic lethal mutations with snR10 deletion, was here identified as a high copy suppressor of the rrp5delta6 temperature-sensitive allele. ROK1 also acts as a low copy suppressor but cannot bypass the cellular requirement for RRP5. Furthermore, we show that suppression by the Rok1p putative RNA helicase rescues the 18S rRNA synthesis defect caused by the rrp5delta6 mutation. PMID- 9848660 TI - Neomycin B inhibits splicing of the td intron indirectly by interfering with translation and enhances missplicing in vivo. AB - The aminoglycoside antibiotic neomycin B inhibits translation in prokaryotes and interferes with RNA-protein interactions in HIV both in vivo and in vitro. Hitherto, inhibition of ribozyme catalysis has only been observed in vitro. We therefore monitored the activity of neomycin B and several other aminoglycoside antibiotics on splicing of the T4 phage thymidylate synthase (td) intron in vivo. All antibiotics tested inhibited splicing, even chloramphenicol, which does not inhibit splicing in vitro. Splicing of the td intron in vivo requires translation for proper folding of the pre-mRNA. In the absence of translation, two interactions between sequences in the upstream exon and the 5' and 3' splice sites trap the pre-mRNA in splicing-incompetent conformations. Their disruption by mutations rendered splicing less dependent on translation and also less sensitive to neomycin B. Intron splicing was affected by neither neomycin B nor gentamicin in Escherichia coli strains carrying antibiotic-resistance genes that modify the ribosomal RNA. Taken together, this demonstrates that in vivo splicing of td intron is not directly inhibited by aminoglycosides, but rather indirectly by their interference with translation. This was further confirmed by assaying splicing of the Tetrahymena group I intron, which is inserted in the E. coli 23 S rRNA and, thus, not translated. Furthermore, neomycin B, paromomycin, and streptomycin enhanced missplicing in antibiotic-sensitive strains. Missplicing is caused by an alternative structural element containing a cryptic 5' splice site, which serves as a substrate for the ribozyme. Our results demonstrate that aminoglycoside antibiotics display different effects on ribozymes in vivo and in vitro. PMID- 9848662 TI - Protein-RNA interactions in the U5 snRNP of Saccharomyces cerevisiae. AB - We present here the first insights into the organization of proteins on the RNA in the U5 snRNP of Saccharomyces cerevisiae. Photo-crosslinking with uniformly labeled U5 RNA in snRNPs reconstituted in vitro revealed five contacting proteins, Prp8p, Snu114p, p30, p16, and p10, contact by the three smaller proteins requiring an intact Sm site. Site-specific crosslinking showed that Snu114p contacts the 5' side of internal loop 1, whereas Prp8p interacts with five different regions of the 5' stem-loop, but not with the Sm site or 3' stem loop. Both internal loops in the 5' domain are essential for Prp8p to associate with the snRNP, but the conserved loop 1 is not, although this is the region to which Prp8p crosslinks most strongly. The extensive contacts between Prp8p and the 5' stem-loop of U5 RNA support the hypothesis that, in spliceosomes, Prp8p stabilizes loop 1-exon interactions. Moreover, data showing that Prp8p contacts the exons even in the absence of loop 1 indicate that Prp8p may be the principal anchoring factor for exons in the spliceosome. This and the close proximity of the spliceosomal translocase, Snu114p, to U5 loop 1 and Prp8p support and extend the proposal that Snu114p mimics U5 loop 1 during a translocation event in the spliceosome. PMID- 9848663 TI - A tabulated review of capillary electrophoresis of carbohydrates. AB - This review summarizes publications on capillary electrophoresis (CE) of carbohydrates, covering almost all hitherto published papers on this topic. It is designed to be a convenient tool for the literature search by providing a comprehensive table. Since CE analysis of carbohydrates is generally complicated due to the structural diversity of carbohydrate species, an attempt is made in this table to supply detailed information on the analyzed form (underivatized or derivatized, type of derivative) and analytical conditions (capillary size, state of the inner wall, composition of the electrophoretic solution, applied voltage, detection method, etc.), for each combination of carbohydrate species to be analyzed. In addition, a brief overview is presented to help in the literature search. PMID- 9848661 TI - Purine-rich enhancers function in the AT-AC pre-mRNA splicing pathway and do so independently of intact U1 snRNP. AB - A rare class of introns in higher eukaryotes is processed by the recently discovered AT-AC spliceosome. AT-AC introns are processed inefficiently in vitro, but the reaction is stimulated by exon-definition interactions involving binding of U1 snRNP to the 5' splice site of the downstream conventional intron. We report that purine-rich exonic splicing enhancers also strongly stimulate sodium channel AT-AC splicing. Intact U2, U4, or U6 snRNAs are not required for enhancer function or for exon definition. Enhancer function is independent of U1 snRNP, showing that splicing stimulation by a downstream 5' splice site and by an exonic enhancer differ mechanistically. PMID- 9848664 TI - A survey of methodological challenges for glycosaminoglycan/proteoglycan analysis and structural characterization by capillary electrophoresis. AB - Proteoglycans participate and regulate several physiological processes via their glycosaminoglycan constituents. For a deeper understanding of how they interact with extracellular ligands as well as with cell bound effector molecules, the fine chemical structures of their glycosaminoglycan chains must be elucidated. Lately developed capillary electrophoretic techniques is a powerful analytical tool for the analysis of glycosaminoglycans, combining a high resolving power with sensitive detection. In this review we describe how depolymerized and intact glycosaminoglycans/proteoglycans can be characterized by capillary electrophoresis, relating these analyses to their possible biological significance. Conditions for running these separations and the detection systems for particular applications are also summarized. PMID- 9848665 TI - Electrophoretic methods for process monitoring and the quality assessment of recombinant glycoproteins. AB - In many ways electrophoretic techniques appear ideal for quality monitoring of proteins and are thus well suited for the analysis of recombinant glycoproteins. The requirements of high throughput, comparative analysis and resolution of many variants are met by several electrophoretic techniques. A wide variety of such techniques are available to biotechnologists in the rapidly developing area of recombinant glycoproteins. It is the aim of this review to specifically cover recent work which has been applied to the analysis of DNA-derived glycoproteins, both from a process control standpoint and final product validation. All major areas of electrophoresis including sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing and techniques utilizing capillary electrophoresis are covered, with emphasis on analysis of glycoforms and oligosaccharide profiles of recombinant glycoproteins. As illustration, actual examples rather than standard glycoproteins are given to indicate the potential and limitations which may be encountered. It is anticipated that this review will prove a useful and practical guide to the latest developments by indicating the relevant merits of different methods. PMID- 9848666 TI - Characterization of sugar chain structures of human alpha-fetoprotein by lectin affinity electrophoresis. ibarahp@oka.urban.ne.jp. AB - Alpha-Fetoprotein (AFP) glycoforms, defined as AFP with different chemical structures of carbohydrate, were analyzed by affinity electrophoresis with several lectins of known specificities against complex-type oligosaccharides. Serum AFP samples from cord blood on full-term delivery and from patients with hepatocellular carcinoma and extrahepatic malignancies including gastrointestinal tumors and yolk sac tumors were used. Two-dimensional lectin affinity electrophoresis and also lectin affinity chromatography coupled with lectin affinity electrophoresis were employed. More than ten AFP glycoforms were identified or characterized using the above-mentioned AFP samples. Known specificities of the lectins against complex-type oligosaccharides were refined or their additional specificities were found in this study. Lectin appeared to have specificity against carbohydrates by recognizing not only specific residues but also the whole carbohydrate molecule containing the residues, resulting in differential affinities for the lectin. PMID- 9848667 TI - Analysis of starch structure using fluorophore-assisted carbohydrate electrophoresis. AB - The analysis of the fine structure of starches is important to the investigation of linkages between starch structure and function and to the investigation of the properties and roles of starch biosynthetic, modifying and degradation enzymes. Fluorophore-assisted carbohydrate electrophoresis has recently been introduced as a method for the analysis of the oligosaccharide populations released by the enzymatic digestion of starches, which has advantages in resolution and sensitivity over previously used methods, and provides the capacity for the facile analysis of oligosaccharide populations on either a molar or mass basis. The use of fluorophore-assisted carbohydrate electrophoresis for the analysis of oligosaccharides is reviewed with particular reference to the choice of label, efficiency of labeling and separation techniques. Examples of separations using slab gel electrophoresis, DNA sequencer analysis and capillary electrophoresis are presented and we conclude that on the basis of resolution and reproducibility, capillary electrophoresis is the method of choice for the separation of oligosaccharides of degree of polymerization from 1 to 100. Examples of isoamylase-debranched starches and glycogens analyzed by capillary electrophoresis are presented. The capillary electrophoresis analysis of starch structure through the analysis of oligosaccharides released by the debranching of limit dextrins derived from starches and glycogens is introduced as a useful diagnostic of starch structure. The potential for future development of novel diagnostics for starch structure using fluorophore-assisted carbohydrate electrophoresis is discussed. PMID- 9848669 TI - Electrophoretic behavior and size distribution of the acidic polysaccharides produced by the bacteria Bradyrhizobium (Chamaecytisus) strain BGA-1 and Bradyrhizobium japonicum USDA 110. AB - The electrophoretic behavior in polyacrylamide gels of the acidic polysaccharides produced by the soil bacteria Bradyrhizobium (Chamaecytisus) strain BGA1 and Bradyrhizobiumjaponicum USDA1 10 has been studied. Both polysaccharides were polydisperse, producing a ladder-like pattern after fixation with Alcian Blue and silver staining of the gel. The polysaccharide molecules were separated according to their size, and they behaved as a collection of flexible random coils of different size and similar charge/mass ratio. The electrophoretic behavior was not affected by the presence of acetyl groups in the polysaccharide. The range of molecular weights of the exopolysaccharide produced by B. japonicum USDA110 was wider and with larger molecules than that of the polysaccharide produced by strain BGA1. The resolution was dependent on the electrophoresis buffer; the best results were achieved with Tris-borate; in Tris-glycine buffer, the resolution was worse, and it was not improved by the addition of sodium dodecyl sulfate (SDS). PMID- 9848668 TI - Structural determination of oligosaccharides from recombinant iduronidase released with peptide N-glycanase F using fluorophore-assisted carbohydrate electrophoresis. AB - The lysosomal storage disorder mucopolysaccharidoses I (MPS I) is caused by a deficiency in the production of alpha-L-iduronidase. Recently, a recombinant alpha-L-iduronidase has been produced in Chinese hamster ovary (CHO) cells. It is thought that for alpha-L-iduronidase to be correctly targeted to the lysosomal vesicle a particular oligosaccharide make-up must be present, and characterization of the carbohydrates is critical. Oligosaccharides from alpha-L iduronidase were analyzed using fluorophore-assisted carbohydrate electrophoresis (FACE). The FACE system uses polyacrylamide gel electrophoresis to separate, quantify, and determine the sequence of oligosaccharides released from glycoproteins. Asparagine-linked oligosaccharides were released from alpha-L iduronidase using the enzyme peptide N-glycanase F (PNGase F). Released oligosaccharides were labeled with a fluorophore at the reducing termini by reductive amination. A total of nine bands were sequenced from the released pool of oligosaccharides. The pool of fluorescently labeled oligosaccharides was then electrophoresed in preparative gels and each band individually excised and extracted. Isolated bands were treated with a series of exoenzymes to determine the sequence of monosaccharides that make up a particular oligosaccharide. A total of eighteen different oligosaccharides were identified from the original pool of oligosaccharides. A majority of the oligosaccharides, over 73%, were found to be of the sialylated complex type. Four of the oligosaccharides were phosphorylated, making up approximately 11% of the carbohydrate pool, and the remaining 15% were of the oligomannose type. PMID- 9848670 TI - Determination of glucose by capillary electrophoresis/laser-induced fluorescence in transdermally collected samples. AB - Capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection has been used for the determination of glucose in samples collected by noninvasive means. The method uses an enzymatic reaction scheme that provides for the determination of small quantities of glucose with detection limits of 80 nM. This approach is used to evaluate passive transdermal diffusion as a noninvasive means to sample glucose in vivo. A simple sampling cell design is presented. Sample collection was performed on volunteer human subjects. Our experiments show that fluctuations in blood glucose concentration are reflected in the samples obtained by passive transdermal diffusion after glucose intake. The results indicate that glucose from the subcutaneous fluid can be accessed by passive diffusion. PMID- 9848671 TI - Investigation of micelles and anionic cyclodextrins as pseudostationary phases for the capillary electrophoresis separation of oligosaccharides derivatized with 2-amino-benzamide. AB - Oligomers of glucose and oligosaccharides released from glycoproteins were derivatized with 2-aminobenzamide. As this fluorophore imparts no charge to the oligosaccharides, several strategies were investigated to achieve capillary electrophoresis (CE) separation of both neutral and charged derivatized glycans. Micellar electrokinetic capillary chromatography (MEKC) with the addition of anionic surfactants was evaluated as a first approach: sodium dodecyl sulfate (SDS) produced the best separation of the oligoglucose fragments, where the migration was inversely related to their degree of polymerization. To demonstrate the applicability of this method for complex carbohydrate analysis, oligosaccharide mixtures derived from ribonuclease B (RNase B) and alpha-acid glycoprotein (alpha-AGP) were analyzed. A satisfactory separation for the high mannose structures found in RNase B could be obtained, whereas charged oligosaccharides from alpha-AGP were poorly resolved. Cyclodextrin-modified CE was chosen as the second approach: the effect of the addition of sulfobutylether beta-cyclodextrin (SBE-beta-CD) or sulfobutylether-gamma-cyclodextrin (SBE-gamma CD) on the electrophoretic mobilities and resolution of neutral and charged oligosaccharides was then studied. Selectivity of sialylated structures could be further improved by using anionic cyclodextrins (CDs) instead of micelles. However, this latter approach failed to baseline-resolve the different high mannose structures of RNase B. A successful separation of the complex mixture of oligosaccharides from alphaalpha-AGP was obtained with the addition of 4% of SBE gamma-CD and triethylamine (TEA) in a phosphate buffer, pH 6.7. PMID- 9848672 TI - Profiling glycoprotein n-linked oligosaccharide by capillary electrophoresis. AB - A method for analysis of N-linked oligosaccharides derived from glycoproteins including sialic acid-containing species is presented. It is based on the combination of specific chemical and enzymatic conversions coupled with capillary electrophoretic (CE) separation and laser-induced fluorescence (LIF) detection. Glycoproteins were heat-denatured in the presence of a reducing agent and the N linked oligosaccharides were released by peptide N-glycosidase (PNGase F; EC3.5.1.52)-catalyzed hydrolysis. The released N-linked oligosaccharides were derivatized with 8-aminopyrene-1,3,6-trisulfonate (APTS) under mild reductive amination conditions in which desialylation and loss of fucose residues are minimized. A model N-linked oligosaccharide, desialylated, galactosylated biantennary, core-substituted with fucose (A2F) was tested for APTS-based derivatization chemistry with excellent recovery of the adduct without losing fucose and neuraminic acid residues. The profiles of heavily sialylated N-linked oligosaccharides derived from fetuin, recombinant human erythropoietin and kallikrein are reported and the data show that the present method produces a high resolution of the N-linked oligosaccharide profile for fingerprinting glycans derived from glycoproteins. PMID- 9848673 TI - Separation of disaccharides by affinity capillary electrophoresis in lectin containing electrophoretic solutions. AB - Separation of the 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatives of simple disaccharides (maltose, cellobiose, gentiobiose, lactose, and melibiose) by affinity capillary electrophoresis was investigated using lectin-containing neutral phosphate buffers, filled in a linear polyacrylamide-coated capillary. When Lens culinaris agglutinin (LCA) was added, the derivatives of glucobioses were retarded with varying magnitudes depending on the amount of LCA and were well separated from each other and from galactosyl glucose under optimized conditions. Addition of Ricinus communis 60 kDa agglutinin (RCA60) to the phosphate buffer gave a different migration profile, in which the derivatives of galactosyl glucoses were more retarded than those of glucobioses. However, addition of either lectin did not accomplish complete separation of the derivatives of all these disaccharides even under optimum conditions. The addition of two kinds of lectins in appropriate proportions improved separation. Thus, the binary system composed of LCA and RCA60, as well as LCA and soybean agglutinin from Glycine max (SBA), gave better separation of these derivatives, giving peak tops for all derivatives. PMID- 9848674 TI - Capillary affinity chromatography and affinity capillary electrophoresis of heparin binding proteins. AB - A new approach for separation, capillary affinity chromatography, is introduced for studying the interaction of heparin with antithrombin III and secretory leukocyte proteinase inhibitor. Heparin is covalently immobilized on the surface of an etched capillary through a silane spacer. The proteins are injected into the heparinized capillary, bound to the heparin, washed with buffer, eluted with sodium chloride in the same buffer using a pressure injection mode and eluting protein detected by absorbance. The resulting affinity separation is similar to that obtained from traditional affinity chromatography. The quantity of loaded protein in capillary affinity chromatography is at the nanogram level, offering an improvement over the milligram levels required for standard affinity chromatographic methods. PMID- 9848675 TI - Peptide capillary zone electrophoresis mass spectrometry of recombinant human erythropoietin: an evaluation of the analytical method. AB - An evaluation of capillary zone electrophoresis-mass spectrometry (CZE-MS) as an analytical methodology for the separation and characterization of complex glycopeptides and nonglycopeptide structures has been performed. The evaluation employed endoproteinase V8 digested recombinant human erythropoietin (rHuEPO) that was further fractionated by reverse phase chromatography. The peptides were subjected to sequence analysis and evaluated by capillary electrophoresis, with or without mass detection, for peptide purity. The peptide mass determined from the sequence was then compared to the mass obtained from CZE-MS. Glycosylation sites and carbohydrate branch patterns were easily determined, site specific microheterogeneity (either O-acetylation of N-acetylneuraminic acids or lactosamine extensions of the carbohydrate chain length) was assessed directly, glycosylation site occupancy was evaluated qualitatively, and nonglycopeptides were resolved and analyzed on-line with ease. Incomplete peptide digestion products were detected and identified by CZE-MS. Protein sequence coverage by CZE MS was 98.2 percent complete from a single map. Off-line evaluation of peptide purity by CZE greatly aided the interpretation of multiple sequence analysis and, in validating that, the CZE-MS was detecting all peptides present. All off-line CZE and on-line CZE-MS experiments employed a capillary that was dynamically coated with Polybrene in the presence of polyethylene glycol; separations were conducted in 0.67 M formic acid. PMID- 9848676 TI - Enhancement of sample loadings for the analysis of oligosaccharides isolated from Pseudomonas aeruginosa using transient isotachophoresis and capillary zone electrophoresis - electrospray - mass spectrometry. AB - The analysis of underivatized core oligosaccharides arising from mild acid hydrolysis of lipopolysaccharides from Pseudomonas aeruginosa serotype 05 was achieved using a transient isotachophoretic preconcentration method coupled to capillary zone electrophoresis-electrospray-mass spectrometry (tCITP-CZE-ES-MS). The combination of a tCITP preconcentration step provided a 10- to 50-fold enhancement of sample loading and a corresponding improvement in sensitivity compared to the conventional zone electrophoresis format. Electrophoretic conditions, enabling the separation of these anionic analytes, were developed to determine possible sites of heterogeneity on either the core or the O-chain structures. The tCITP-CZE-ES-MS technique provided unparalleled resolution of the different core glycoforms and oligosaccharides obtained from the acid cleavage of the native endotoxins whether isolated following conventional gel permeation chromatography or obtained from direct hydrolysis of the bacterial isolates. These investigations also highlighted the highly phosphorylated nature of these complex cell membrane components, where the heptose residues of the core oligosaccharide can bear up to six phosphate groups. PMID- 9848677 TI - Isolation and characterization of beta-cyclodextrin sulfates by preparative gradient polyacrylamide gel electrophoresis, capillary electrophoresis and electrospray ionization - mass spectrometry. AB - A beta-cyclodextrin sulfate mixture has been fractionated using discontinuous gradient polyacrylamide gel electrophoresis. Semidry electrotransfer of the sample onto a positively charged nylon membrane and visualization of a portion of this membrane with Alcian blue stain showed multiple bands. The bands were cut from the remaining portion of the membrane and after washing with 8 M urea, the beta-cyclodextrin sulfate fractions were eluted with 2 M sodium chloride and dialyzed. Analysis of each fraction using high resolution analytical gradient polyacrylamide gel electrophoresis as well as capillary electrophoresis, using indirect detection, showed some of the fractions to be pure while others were mixtures. Each beta-cyclodextrin sulfate fraction was complexed with a basic synthetic peptide and analyzed by electrospray ionization mass spectrometry to define the mass of the components in each mixture and thereby to determine the purity of each sample. PMID- 9848678 TI - Separation of 1-phenyl-3-methyl-5-pyrazolone derivatives of monosaccharides by capillary electrochromatography. AB - 1-Phenyl-3-methyl-5-pyrazolone (PMP) derivatives of component monosaccharides in glycoproteins (fucose, galactose, mannose, N-acetylgalactosamine and N acetylglucosamine) and epimeric aldopentoses (arabinose, lyxose, ribose and xylose) were well separated from each other by capillary electrochromatography on a Hypersil ODS column with a mixture of 50 mM N-(2-hydroxyethyl)piperazine-2'-(2 ethanesulfonic acid) buffer, pH 6.0 to approximately 6.3, and acetonitrile (2.2:1 v/v) as eluent. The elution of these compounds showed relatively strong dependence on the pH and concentration of the buffer salts contained in the eluent, as compared to the elution by pressure-driven high-performance liquid chromatography (HPLC) on the same stationary phase, but separation of PMP monosaccharides was better than that by HPLC. Retention times of PMP monosaccharides were highly reproducible with a relative standard deviation (RSD) of approximately 0.6%, and quantification with an RSD less than 5% could be achieved using 3-O-methylglucose as an internal standard. PMID- 9848679 TI - Cholera toxin differentially regulates nitric oxide synthesis, tumor necrosis factor-alpha production and respiratory burst in murine macrophages. AB - The aim of this study was to determine if cholera toxin can modulate the expression of several macrophage effector functions. The effect of cholera toxin on the induction of NO synthesis, production of tumor necrosis factor-alpha and induction of respiratory burst was examined in the J774.A2 macrophage cell line. Pre-incubation of cell cultures with cholera toxin significantly down-regulated lipopolysaccharide-induced NO synthesis and phorbol-12-myristate-13-acetate induced respiratory burst. Concomitant addition of cholera toxin and lipopolysaccharide to cell cultures enhanced the production of tumor necrosis factor-alpha. These effects were abrogated when cholera toxin was inactivated by heat or treated with a specific monoclonal antibody. PMID- 9848680 TI - Th2 biased immune response in cases with active Mycobacterium tuberculosis infection and tuberculin anergy. AB - This study was aimed at investigating the immunologic relationship between cytokine production pattern and tuberculin negativity in patients with active Mycobacterium tuberculosis infection. After classifying patients by the extent of pulmonary involvement and the size of the tuberculin reaction, we evaluated the rate of cytokine positivity in peripheral blood to determine whether there is a characteristic cellular immune reaction pattern which could partly explain the tuberculin negativity in some of these cases. The significance of tuberculin anergy occurring in some cases with M. tuberculosis infection is still not clear. We investigated the ratio of IL- 4, IL-10, IL-12, CD-4, CD-8 expressing lymphocytes in the peripheral blood of patients with active M. tuberculosis infection and correlated the percentage of the reactive cells with the positivity or negativity of tuberculin skin reactions. Twenty-eight patients were included in the study, with 11 healthy volunteers serving as controls. 10 ml of venous blood was drawn before starting anti-mycobacterial treatment. A tuberculin skin test was performed, introducing intracutaneously 5 TU PPD on the forearm with results evaluated after 72 h. Consistent with the reactivity or non-reactivity of the tuberculin skin test, we found a significantly higher ratio of IL-4 and IL-10 positive lymphocytes and a significantly lower ratio of IL-12 in the peripheral blood of patients with tuberculin anergy than in that of tuberculin positive patients or healthy donors. There was no difference in the ratio of the CD-4 CD-8 positive lymphocytes among the three groups. To evaluate whether the differences could be explained by the degree of pulmonary tubercular involvement, we classified the patients into three groups according to the extent and type of X ray findings. Seven out of eight tuberculin negative patients were classified as grade III, whereas in the tuberculin positive group only seven out of 20 fell in this category. There was no significant correlation between the radiological grade of the patients and the examined in vitro parameters unless the tuberculin reactivity of each patients was also considered. Tuberculin anergy may reflect an inappropriate immune response to the intracellular pathogen. The high percentage of IL-4 and IL-10 positive lymphocytes together with a low percentage of IL-12 positive lymphocytes in the peripheral blood of anergic patients suggests a Th2 biased immune response during the early course of the disease. PMID- 9848681 TI - Isolation of recombinant phage clones expressing mycobacterial T cell antigens by screening a recombinant DNA library with human CD4+ Th1 clones. AB - A lambda gt11 recombinant DNA library of Mycobacterium leprae was screened to isolate recombinant phage clones expressing mycobacterial antigens important for T cell reactivity. The library was plated on a lawn of Escherichia coli Y1090 and recombinant antigens were expressed from isolated phage clones in 96-well plates. Pools of recombinant antigens from 12 wells were tested in T cell proliferation assays with MHC class II restricted human CD4+ Th1 clones secreting interferon gamma and cytotoxic for antigen pulsed antigen presenting cells. By screening 1750 pools of recombinant antigens with a mixture of eight Th1 clones, we identified two recombinant phage clones that expressed recombinant mycobacterial antigens stimulatory for T cells. MHC restriction analysis and reactivity to a battery of mycobacterial antigens suggested that the two responding Th1 clones recognized mycobacterial antigens/epitopes with different MHC class II (HLA-DR) restriction requirements. Our results suggest that the methodology described in this paper is suited to isolate recombinant phage clones expressing mycobacterial recombinant antigens stimulatory for T cells of protective phenotype. Such antigens may be useful in designing new vaccines and diagnostic reagents against mycobacterial diseases. PMID- 9848682 TI - Non-random fragmentation of ribosomal RNA in Helicobacter pylori during conversion to the coccoid form. AB - The integrity of DNA and ribosomal RNAs in exponentially growing (bacillary) and ageing stationary phase (coccoid) cultures of Helicobacter pylori type strain CCUG 17874 was investigated. Extensive non-random fragmentation of rRNAs was observed during the conversion to the coccoid form. Beside a small proportion of full-length 16S and 23S rRNA that was always present, the majority of both 16S and 23S rRNA molecules showed distinct highly specific fragmentation patterns. The 16S rRNA fragmentation was characterised in detail by means of Northern blot and primer extension analysis. One cleavage site was located within the highly conserved U5 region (position about 920). The results could not be attributed to the presence of intervening sequences in the 16S and 23S rRNA genes. PMID- 9848683 TI - Retention of neutralising activity by recombinant anti-pneumolysin antibody fragments. AB - The variable domains of a neutralising (prevents erythrocyte lysis) anti pneumolysin monoclonal antibody have been cloned and expressed as functional protein in Escherichia coli. Purification of the anti-pneumolysin single-chain antibody fragment, via antibody-affinity or metal-chelate affinity chromatography, resulted in product that was predominantly in a dimeric or monomeric form, respectively. The dimeric single-chain antibody fragment showed a higher sensitivity and affinity for immobilised antigen in both ELISA and BIAcore studies. The dimeric single-chain antibody fragment was as effective at protecting erythrocytes from lysis as the parent monoclonal. The monomeric, low affinity single-chain antibody fragment, showed reduced neutralising potency. As antibiotic resistant Streptococcus pneumoniae strains continue to show an increasing word-wide distribution, recombinant, neutralising antibody fragments, may provide an additional class of molecules useful in the treatment of toxaemia. PMID- 9848684 TI - A highly sensitive and fast non-radioactive method for the detection of polymerase chain reaction products from Salmonella serovars, such as Salmonella typhi, in blood specimens. AB - A polymerase chain reaction based test was developed for the detection of Salmonella spp. in blood specimens. After amplification of a 389 bp-polymerase chain reaction product from the invA gene, a microtiter plate hybridization assay was performed. The protocol described allowed the detection of six to seven copies of the Salmonella typhi genome, as determined by serial dilutions of DNA from S. typhi. Eighteen blood specimens from artificially infected rats and 22 blood specimens from patients were analyzed to validate the method. Considering that the most frequent Salmonella serovar isolated from blood in case of bacteremia is S. typhi, the polymerase chain reaction-microtiter plate hybridization technique could be used as a novel, rapid diagnostic method for typhoid fever, particularly when standard culture assays are negative. PMID- 9848685 TI - Detection of Mycoplasma salivarium and Mycoplasma fermentans in synovial fluids of temporomandibular joints of patients with disorders in the joints. AB - Thirty-six synovial fluid samples of temporomandibular joints were obtained from 33 patients with pain and anterior disk displacement (closed lock) in the joints. DNAs were prepared from the samples and amplified by a PCR-based assay specific for Mycoplasma salivarium or Mycoplasma fermentans. Of the 36 samples, five (14%), three (8%), and 19 (53%) were positive for M. salivarium, M. fermentans and both, respectively. PMID- 9848686 TI - Gamma globulin, Evan's blue, aprotinin A PLA2 inhibitor, tetracycline and antioxidants protect epithelial cells against damage induced by synergism among streptococcal hemolysins, oxidants and proteinases: relation to the prevention of post-streptococcal sequelae and septic shock. AB - An in vitro model was employed to study the potential role of streptococcal extra cellular products, rich in streptolysin O, in cellular injury as related to streptococcal infections and post-streptococcal sequelae. Extra-cellular products (EXPA) rich in streptolysin O were isolated from type 4, group A hemolytic streptococci grown in a chemostat, in a synthetic medium. EXPA induced moderate cytopathogenic changes in monkey kidney epithelial cells and in rat heart cells pre-labeled with 3H-arachidonate. However very strong toxic effects were induced when EXP was combined with oxidants (glucose oxides generated H2O2, AAPH-induced peroxyl radical (ROO.), NO generated by sodium nitroprusside) and proteinases (plasmin, trypsin). Cell killing was distinctly synergistic in nature. Cell damage induced by the multi-component cocktails was strongly inhibited either by micromolar amounts of gamma globulin, and Evan's blue which neutralized SLO activity, by tetracycline, trasylol (aprotinin), epsilon amino caproic acid and by soybean trypsin inhibitor, all proteinase inhibitors as well as by a non penetrating PLA2 inhibitor A. The results suggest that fasciitis, myositis and sepsis resulting from infections with hemolytic streptococci might be caused by a coordinated 'cross-talk' among microbial, leukocyte and additional host-derived pro-inflammatory agents. Since attempts to prolong lives of septic patients by the exclusive administration of single antagonists invariably failed, it is proposed that the administration of 'cocktails' of putative inhibitors against major pro-inflammatory agonizes generated in inflammation and infection might protect against the deleterious effects caused by the biochemical and pharmacological cascades which are known to be activated in sepsis. PMID- 9848687 TI - Fibronectin in saliva inhibits Porphyromonas gingivalis fimbria-induced expression of inflammatory cytokine gene in mouse macrophages. AB - The aim of this study was to examine whether fibronectin in saliva plays a regulatory role in Porphyromonas gingivalis fimbria-mediated pathogenesis in adult periodontal disease. Our previous study demonstrated that fibronectin is one of the binding proteins of P. gingivalis fimbrillin. In the present study, we observed that fibronectin in saliva binds specifically to the fimbrillin. The fibronectin content in saliva of adult periodontal patients was significantly lower than that of healthy subject. In addition, the inhibitory action of salivary fibronectin from adult periodontal patients toward P. gingivalis fimbria induced expression of the neutrophil chemoattractant KC gene in mouse macrophages was clearly weak compared with that of the fibronectin from healthy subjects. These results suggest that fibronectin in saliva plays an important role as a regulator in the pathogenesis of P. gingivalis fimbriae in adult periodontal disease. PMID- 9848688 TI - Comparison of the effects of Salmonella minnesota Re595 lipopolysaccharide, lipid A and monophosphoryl lipid A on nitric oxide, TNF-alpha, and IL-6 induction from RAW 264.7 macrophages. AB - Lipopolysaccharide (LPS) exhibits a wide variety of bioactivities. Although it was generally proposed that the lipid A component represented the active center responsible for most of the bioactivities of LPS, a variety of lipid A partial structures and analogues were reported to have different properties. Lipopolysaccharide of the Re595 mutant of Salmonella minnesota is lack of O and part of the core polysaccharide (2 keto-3-deoxyoctanate (KDO) left on lipid A). Re595 lipid A (LA) and Re595 monophosphoryl lipid A (MPLA) differ in structure from Re595 LPS by lacking KDO and KDO plus phosphoryl group respectively. Whether these lipid A-common Re595 LPS preparations differed in activities, we investigated their effects on nitric oxide (NO), TNF-alpha, IL-6, and IL-12 induction from murine macrophage cell line RAW 264.7. RAW 264.7 cells (2 x 10(5) cells ml(-1)) were stimulated with these LPS preparations at 1 microg ml(-1) for 48 h. Re595 LPS, Re595 LA and Re595 MPLA significantly induced NO, TNF-alpha and IL-6 production; NO, TNF-alpha and IL-6 inducing capacities were in the order of LPS = LA > MPLA, LPS = LA = MPLA, and LPS = LA > MPLA respectively. However, these preparations did not induce IL-12 production from RAW cells even when stimulated in combination with IFN-gamma (20 U ml(-1)). IFN-gamma itself also induced NO, TNF-alpha and IL-6 production from RAW 264.7 cells. When RAW 264.7 cells were stimulated with IFN-gamma plus any of these preparations, effects were additive and synergistic for NO and IL-6 responses respectively. But TNF-alpha responses of RAW cells against these preparations were almost equal when cultured alone or in combination with IFN-gamma. Pre-treatment of RAW cells either with LPS, LA or MPLA at low concentration (0.1 microg ml(-1)) for 60 min before pulsing with IFN-gamma (20 IU ml(-1)) plus LPS (1 microg ml(-1)) for an additional 48 h, significantly (P < 0.01) decreased NO response. Although to a lesser extent, TNF-alpha and IL-6 responses were also decreased. Complete inhibition of NO inducing effect of these LPS preparations was achieved with polymyxin B at 40 microg ml(-1). But the concentration of polymyxin B to get a significant (P < 0.05) inhibitory effect on LPS was four times higher than that for LA or MPLA. Unexpectedly, polymyxin B also inhibited INF-gamma-induced NO production from RAW cells in a dose-dependent fashion. These findings suggested that effect of LPS was dependent, at least in part, on both the LPS polysaccharide chain length and the hydrophilic portion of LPS. In addition, not only LPS but also LA and MPLA exert either enhancing or suppressive effects, depending on their concentrations and the timing of their addition with respect to co-stimulators. PMID- 9848689 TI - Pre-existing diabetics: relationship between glycemic control and emotional status in pregnancy. AB - Due to the paucity of information regarding the emotional status of pregnant diabetic women, we sought to determine the impact of pre-existing diabetes on the relation between the level of glycemic control and emotional well-being during pregnancy. English-speaking women with pre-existing diabetes mellitus (N = 100) and nondiabetic controls (N = 132) were compared for maternal characteristics and test results on the Profile of Mood States-Bipolar form. It is a 72-item, self administered Likert scale measuring six bipolar subjective mood states representing both positive and negative aspects. Patients were stratified according to level of glycemic control. The number of glucose determinations were categorized as: mild and severe hypoglycemia and hyperglycemia, and normoglycemia. An Average Mood Disturbance score was calculated to measure the level of association between the total psychological profile and categories of glucose determinations. In two of the six subscales (hostility and anxiety) of the Profile of Mood States, significantly lower scores indicating greater hostility and anxiety were found in the diabetic group. The level of glycemic control was not adversely associated with the emotional state of pre-existing diabetic pregnant women. The psychological profile was not associated with the severity of the disease as reflected by the diabetes classification. Pregnant women with pre-existing diabetes display greater anxiety and hostility in comparison to nondiabetic women with no association to level of glycemic control. This issue should be addressed in the medical and psychosocial health assessment of these women. PMID- 9848690 TI - Informed consent and the redefining of conception: a decision ill-conceived? AB - The purposes of this study are to assess the use of the American College of Obstetricians and Gynecologists' (ACOG) definitions of conception (a synonym for implantation) and the beginning of pregnancy (at implantation) in the clinical practice of its members and to explore the implications of differing definitions of conception and pregnancy onset for the process of informed consent. A survey was mailed to 112 members of the Louisville Ob/Gyn Society asking what definition of conception they used in their clinical practice and when they judged pregnancy began. A second mailing was sent to nonresponding members. Using logistic regression analysis, the responses to these questions were evaluated with respect to practice type, number of years in practice, and the ACOG membership. Responses were received from 86% (96 of 112) of the members. A total of 73% (70 of 96) (95% CI 69-77%) of the members indicated that conception was a synonym for fertilization, and 24% (23 of 96) (95% CI 21-28%) indicated that conception was a synonym for implantation (P < .001). Of the members, 50% (48 of 96) indicated that pregnancy began at fertilization, and 48% (46 of 96) indicated that pregnancy began with implantation (NS). Regression analysis failed to demonstrate a significant relationship to type of practice, years in practice, or the ACOG membership for these responses. Neither ACOG definition has been consistently adopted by its members whose definitions are more consistent with lay and embryologist definitions. Potentially, the process of informed consent is jeopardized by these ambiguities. The ACOG is urged to reconsider its definitions. PMID- 9848691 TI - Relationship between endocervical canal length between 15-24 weeks gestation and obstetric history. AB - The object was to determine whether there is a correlation between the obstetric history and the ultrasonographically determined endocervical canal length between 15 and 24 weeks gestation. A retrospective cohort study was performed in singleton pregnancies of multigravidas with normal and abnormal obstetric histories. They underwent sonographic evaluation for the determination of the endocervical canal length between 15 and 24 weeks gestation. The shortest endocervical canal length measurements between 15 and 20 weeks and also between 21 and 24 weeks of gestation were recorded. An ultrasound diagnosis of cervical incompetence was defined as progressive shortening of the endocervical canal length to <2 cm or a single endocervical canal length measurement <2 cm. A multivariable general linear regression model was used to correlate the relationship between endocervical canal lengths at 15-20 weeks and 21-24 weeks gestation in the current pregnancy with the earliest gestational age at delivery of prior pregnancies. Chi-square test was used to determine the relationship between the development of an ultrasound diagnosis of cervical incompetence and the earliest gestational age at delivery of prior pregnancies. A total of 155 pregnancies were studied. The number of women according to the obstetric history categories were: 57 had delivered <24 weeks, 12 between 24 and 26 weeks, 16 between 27 and 32 weeks, 16 between 33 and 36 weeks, and 54 delivered > or =37 weeks. There was a significant correlation between the endocervical canal length measurements between 15-20 (P < 0.0001) weeks and 21-24 weeks (P < 0.0001) in the studied pregnancy and the earliest gestational age at delivery of prior pregnancies. A significant relationship between the ultrasound diagnosis of cervical incompetence and the obstetric history category (P = 0.0026) was observed. There were 36 cases of ultrasound diagnosed cervical incompetence with 91.7% (33/36) occurring in women who had a prior <27 weeks' gestation delivery. These data provide further evidence that cervical incompetence is a relative condition and not an "all or none" phenomenon. In addition, women with a prior delivery <30 weeks gestation should be followed with second trimester serial cervical sonography to rule out cervical incompetence. PMID- 9848692 TI - Survey of maternal-fetal medicine subspecialists: professional activities, job setting, satisfaction, and trends over time. AB - The purpose of this survey was to describe the job setting, professional activities, and satisfaction of maternal-fetal medicine (MFM) subspecialists and to compare these with previous surveys. A questionnaire was mailed to members of the Society of Perinatal Obstetricians (SPO). The results were compared to a similar survey in 1986. Of 1,352 members, 58% responded. In comparison to 1986, there was an increase in percentage of women (25%); a decrease in university hospital-based faculty (55%); and an increase in community hospital-based salaried physicians (23%) and private practice (15%). The distribution of professional time changes with an increase in clinical care, a decrease in teaching and research. The annual number of procedures increased for ultrasound and genetics and decreased for gynecologic procedures. Since 1986, there have been significant changes among MFM subspecialists in job setting, allocation of professional time, and number and types of procedures. Job satisfaction remains high. PMID- 9848693 TI - Thrombophilia: a mechanism of disease in women with adverse pregnancy outcome and thrombotic lesions in the placenta. AB - The purpose of this study was to examine the relationship among adverse pregnancy outcome, the presence of thrombotic lesions in the placenta, and the frequency and type of laboratory abnormalities consistent with the presence of a thrombophilic state. A retrospective cohort study was designed to determine the frequency of laboratory abnormalities consistent with thrombophilia among patients with thrombotic lesions of the placenta and adverse pregnancy outcome. The workup for a thrombophilic state included anticardiolipin antibodies, lupus anticoagulant, protein C and antithrombin III activities, protein S total and free, activated protein C resistance ratio, and Factor V Leiden mutation. Thrombotic lesions were identified by histopathologic examination of the placenta. Thirteen patients met the study criteria over an 11-month period. Seven patients were heterozygous for Factor V Leiden mutation (53.8%). Protein S deficiency was found in three cases (23.0%), and no hemostatic abnormality was detected in three cases (23.0%). Mothers with an adverse pregnancy outcome and thrombotic lesions of the placenta often have laboratory abnormalities indicative of a thrombophilic state. We propose that thrombophilia leading to thrombosis in the maternal and/or fetal circulations is a significant mechanism of disease during pregnancy. PMID- 9848694 TI - Experience with intrauterine transfusions for severe Rh alloimmunization in a developing country. AB - This study reports our experience with 67 intrauterine transfusions (IUTs) carried out for 27 cases of severe Rh alloimmunization, which could be useful to other developing countries with similar situations. Most of the mothers were from sections of India other than Mumbai, their socioeconomic status was low, and they were referred during the second or third trimester. The mean gestation age at first IUT was 27+/-2.9 weeks and maternal anti-D titer ranged from 1:32 to 1:512. Ultrasonography (USG) was normal in eight cases, but showed minimal or gross ascites in 8 and 11 cases, respectively. The mean +/- SD hematocrit (HCT) in three groups defined by USG was 23.5+/-1.7, 15.9+/-4, and 12+/-5.9, respectively. Amniotic fluid analysis, which proved to be an important investigation, indicated IUT in eight cases having normal USG. Six cases were severely anemic (Hb deficit >7 g/dl). By fetal cell staining, the percentage of the donor's red cells in the fetal circulation was determined. Besides Hb, blood group, direct antiglobulin test, and mean cell volume, this parameter was also useful in assessing efficacy of IUT and the need for an exchange transfusion after birth. Of 11 fetuses having gross ascites, eight and one each from the remaining two groups, were stillborn. One death may be procedure related. Two neonates died due to hemorrhagic disorder and prematurity. The overall survival rate was 55.6%. Late referral, severe Rh alloimmunization, volume overload, delay in IUT because of nonavailability of blood and use of nonirradiated blood could be the reasons for the poor outcome. Strategies for improving results are discussed. PMID- 9848695 TI - Anthropometric differences in macrosomic infants of diabetic and nondiabetic mothers. AB - The objective was to investigate the hypothesis that anthropometric and body composition differences exist between macrosomic infants of diabetic and nondiabetic mothers. Sixteen infants of mothers with diabetes, along with 58 control infants, were studied within 24 hours of delivery. The following measurements were obtained: birthweight, birth length and extremity length; circumferences of the head, chest, shoulders, and extremities; and triceps, subscapular, flank, and thigh skinfolds. Estimation of fat mass and calculation of percent body fat was performed according to the Dauncey method. Macrosomic infants of diabetic mothers were characterized by larger shoulder and extremity circumferences, a decreased head-to-shoulder ratio, significantly higher body fat, and thicker upper extremity skinfolds compared with nondiabetic control infants of similar birthweight and birth length. Differences in body composition and weight distribution may explain the propensity for shoulder dystocia in the diabetic population. PMID- 9848696 TI - In utero allotransplantation of retrovirally transduced fetal hepatocytes in primates: feasibility and short-term follow-up. AB - In utero allotransplantation of fetal hepatocytes into a preimmune fetus could be used in early treatment of many inherited hepatic metabolic diseases. This study was designed to assess the tolerance to hepatocyte transplantation and to test the feasability and toxicity of such an injection in a primate model. Fetal hepatocytes were obtained from two 120-day-old Macaca mulatta fetuses and cryopreserved. They were thawed, cultured in vitro, and transduced with a recombinant retrovirus expressing beta-galactosidase. Transduction efficiency was 75-85%. Three unrelated fetuses (90, 100, and 104 days old) were each given 1-2 x 10(7) transduced cells via the umbilical vein. This caused vasospasm and severe bradycardia. Two fetuses died in the 48 hours after transplantation; the third survived and was killed at the end of gestation. No evidence of the infused cells was found. Three fetuses (90 days old) were, therefore, given 3-4 10(7) hepatocytes by direct intrahepatic injection. All the fetuses survived without side effect. Donor cells were not apparent from histochemical staining and PCR reactions. There was no evidence of inflammatory reaction. These findings indicate that the protocole could be improved by increasing the number of transplanted cells and using specific hepatic promoters in the retroviral vectors to achieve an effective postnatal chimerism. PMID- 9848697 TI - Abnormal maternal serum alpha fetoprotein and pregnancy outcome. AB - The objective was to assess the occurrence of miscarriages, low birth weight, and karyotype abnormalities found with low and elevated maternal serum alpha fetoprotein (MSAFP) among women who had genetic amniocentesis performed. A retrospective study of 2,159 women who had MSAFP analysis prior to amniocentesis was conducted. Pregnancy outcomes were obtained from record review and physicians follow-up. Limits of MSAFP used in analysis were <0.5 adjusted multiples of the median (MOM) (lower levels) and >2.0 MOM (upper levels). Autosomal trisomy was found in 1.6% with low, 0.9% normal, and 0.6% with elevated MSAFP values. Sex chromosome abnormalities were present only in patients with normal MSAFP, [45X (n = 6), 47XXY (n = 2), 69XXX]. Of five open neural tube defects, four had elevated MSAFP and one had a normal value. Omphalocele was identified in four patients, two with normal and two with elevated MSAFP. Gastroschisis was found in one low and one elevated MSAFP. Amniotic fluid alpha-fetoprotein (AFAFP) values did not correlate with MSAFP values. Patients with low MSAFP levels had a greater prevalence of abnormal karyotype (19 of 249, prevalence = 0.076) than patients with an elevated MSAFP level (2 or 166, prevalence = 0.012 OR (odds ratio) = 0.20 (P value = 0.024) when unadjusted for maternal age, and OR = 0.09 (P value = 0.001) when adjusted for maternal age. Spontaneous abortion occurred more often in patients with elevated (4 of 166, or 4%) than normal or low (20 of 1948, or 1%) values of MSAFP (odds ratio 4.32, P = 0.020 when adjusted for maternal age). Birth weight below 2,500 g was present less frequently with low or normal MSAFP (136 of 1,760, or 7.7%) than in elevated MSAFP (21 of 144 or 14.6%) (odds ratio 2.04, P = 0.005, unadjusted; and odds ratio = 2.32, P = 0.003, adjusted for maternal age). Female fetuses were present more often with low MSAFP (136 of 249, or 55%) than elevated levels 43% (71 of 164, or 43%; P = 0.024). We conclude that patients undergoing genetic amniocentesis with MSAFP <.5 MOM are less likely to miscarry, deliver a low birth weight newborn, or have a male infant than patients with MSAFP levels >2.0 MOM. PMID- 9848698 TI - Brown-Sequard syndrome due to isolated blunt trauma. AB - Blunt trauma without associated fracture or ligamentous injury is a rare cause of Brown-Sequard syndrome. We report a case of Brown-Sequard syndrome after a direct blow to the cervical spine that did not cause injury to adjacent bone or ligaments. Characteristic neurologic findings, including a unilateral hemiparesis with associated contralateral sensory findings, were noted at the time of presentation. High-dose steroids were instituted after recognition of the patient's injury, and magnetic resonance imaging of the cervical spine revealed a unilateral cord contusion with no associated fractures. After 1 month, the patient had recovered much of his function and was able to ambulate unassisted. PMID- 9848700 TI - Rupture of ectopic pregnancy after medical therapy with methotrexate: a case series. AB - Medical treatment of ectopic pregnancy with methotrexate has become the standard of care in many areas of the U.S. Rupture of ectopic pregnancy after medical treatment presents a diagnostic challenge to the Emergency Physician. We review a series of 11 patients with ectopic pregnancy treated with methotrexate who then required surgical treatment for rupture. PMID- 9848699 TI - Leptospirosis in an urban setting: case report and review of an emerging infectious disease. AB - Leptospiosis is a common zoonosis affecting most mammals. Leptospirosis has protean manifestations ranging from a flu-like illness to fulminant hepatic and renal failure culminating in death. Although the diagnosis is often not considered upon presentation, the literature suggests that leptospirosis is a reemerging infectious disease in urban centers throughout the industrialized world. It will be incumbent upon Emergency Physicians to include this spirochetal disease in the differential diagnosis of febrile patients with appropriate risk factors and symptomatology. We present the case of a 36 year-old woman who presented to the Emergency Department with fever and hypotension. We review the literature on leptospirosis with specific focus on risk factors and pathogenesis, clinical manifestations, diagnosis, treatment, and outcome. PMID- 9848701 TI - Brillo pad crack screen aspiration and ingestion. AB - Crack cocaine is commonly smoked in a pipe with a metallic filter made from a steel wool scouring pad. We report an unusual complication of smoking crack cocaine: the aspiration and ingestion of a Brillo pad filter. A 34-year-old female presented 7 h after drinking beer and smoking crack. She was concerned that she might have inhaled the "screen" from her crack pipe, a piece of Brillo pad the size of her fingertip. She complained of "burning" in her throat, a foreign body sensation, and change in her voice, but no dyspnea, dysphagia, or abdominal pain. On physical examination, she was afebrile with a pulse of 105 beats/min and respiratory rate of 24 breaths/min. She was tearful and spoke in a whisper. There were no visible oropharyngeal burns and the lungs were clear to auscultation, but she had intermittent inspiratory stridor. The O2 saturation was 96%, and the ethanol concentration was 100 mg/dl. No foreign body or burn was seen on indirect laryngoscopy. A lateral neck x-ray study showed a normal epiglottis and no foreign body. Chest x-ray studies were unremarkable. Fiberoptic laryngoscopy showed left posterior arytenoid edema and swelling. An abdominal x ray study revealed a foreign body in the right lower quadrant consistent with the Brillo pad filter. The next morning, the patient was asymptomatic and was discharged, recovering without sequellae. While crack pipe screen aspiration is a rarely reported event, physicians should be aware of the potential for foreign body aspiration and ingestion by this mechanism. PMID- 9848702 TI - Ultrasonography by emergency physicians in patients with suspected ureteral colic. AB - We performed a prospective study of patients with suspected ureteral colic to evaluate the test characteristics of bedside renal ultrasonography (US) performed by emergency physicians (EPs) for detecting hydronephrosis, and to evaluate how US can be used to predict the likelihood of nephrolithiasis. Thirteen EPs performed US, recorded the presence of hydronephrosis, and made an assessment of the likelihood of nephrolithiasis. All patients underwent i.v. pyelography (IVP) or unenhanced helical computed tomography (CT). There were 126 patients in the study: 84 underwent IVP; 42 underwent helical CT. Test characteristics of bedside US for detecting hydronephrosis were: sensitivity 72%, specificity 73%, positive predictive value (PPV) 85%, negative predictive value (NPV) 54%, accuracy 72%. The PPV and NPV for the ability of the EP to predict nephrolithiasis after performing US were 86% and 75%, respectively. We conclude that bedside US performed by EPs may be used to detect hydronephrosis and help predict the presence of nephrolithiasis. PMID- 9848703 TI - Evaluation of a portable blood gas analyzer for pediatric interhospital transport. AB - We evaluated a portable blood gas analyzer for its speed, reliability, and usefulness during interhospital transportation of critically ill children in a prospective study. The accuracy of a portable blood gas analyzer (PBGA) was first established by comparing its results with values obtained from a standard blood gas analyzer. The speed, accuracy, and usefulness of the PBGA were then compared with those of standard analyzers at 10 referring hospitals during interhospital transportation of critically ill children. A highly significant linear correlation was demonstrated between values obtained using the PBGA and those derived from standard analyzers. The time required to obtain blood gas results was 2 min with the portable device, significantly less than the mean of 8.4 min +/- 6.4 min (range 1-24 min) required to get results from the laboratory facilities of the referring hospitals. Modification of treatment or adjustment to mechanical ventilation was required in 30% of transported patients based on blood gas results obtained immediately before departure from the referring hospital. We conclude that a portable blood gas analyzer provides rapid, reliable, and useful data that help to determine therapy for critically ill children awaiting interhospital transport. PMID- 9848704 TI - Influence of absorbable dusting powders on wound infection. AB - Surgical glove dusting powders are commonly used as mold-release agents and to facilitate donning. Cornstarch and CaCO3 are commonly used absorbable dusting powders. This experimental study demonstrates that these absorbable dusting powders significantly potentiate bacterial growth and enhance a wound's inflammatory response. The infection-potentiating effects of CaCO3 are significantly greater than those of cornstarch. PMID- 9848705 TI - Phenformin and lactic acidosis: a case report and review. AB - Phenformin was removed from the U.S. market 20 years ago because of a high incidence of lactic acidosis. Unfortunately, this medication is still available from foreign sources. Another biguanide, metformin, was reintroduced to the United States market for the treatment of diabetes. Biguanide-induced lactic acidosis should be included in the differential diagnosis of elevated anion gap metabolic acidosis. We present a case of phenformin-induced lactic acidosis in which we were consulted at the local poison control center. We also review its pathophysiology, presentation, and treatment. A review of the actions of phenformin illustrates the mechanism of pathology that may also occur with metformin. Risk factors for the development of lactic acidosis include renal deficiency, hepatic disease, cardiac disease, and drug interaction such as cimetidine. PMID- 9848706 TI - Altered mental status and shock. PMID- 9848707 TI - Perforation of the gall bladder. PMID- 9848708 TI - The University HealthSystems Consortium: a bureaucracy designed to smother academic excellence. PMID- 9848709 TI - Reduction of redundant laboratory orders by access to computerized patient records. AB - From a convenience sample of 500 consecutive patients seen in the emergency department (ED), occasions were recorded when data from the Veterans Affairs Decentralized Hospital Computer Program provided immediate clinical decision support and obviated redundant laboratory tests. Patient care was improved by access to inpatient discharge summaries in 85 cases (19%), laboratory results in 34 (7%), pharmacy records of allergies and prescriptions in 30 (6%), radiologic reports in 19 (4%), and electrocardiograms in 11 (2%). Overall savings in tests, prescriptions, admissions, and errors were estimated at about $5 per visit. Availability of previous laboratory results clearly decreased ordering of redundant studies. Computer-based medical records also provided details of previous diagnoses, treatments, allergies, and current medications. On many occasions, the presumptive diagnosis had already been worked up and proven or disproven, thus simplifying the entire encounter. PMID- 9848710 TI - Reverse transcriptase-polymerase chain reaction analysis of thyrocyte-relevant genes in fine-needle aspiration biopsies of the human thyroid. AB - Currently, fine-needle aspiration cytology is a valuable tool in the routine diagnosis of suspicious thyroid nodules. We present a very sensitive method for the molecular analysis of the expression of several genes important for normal thyroid function in parallel to the cytological diagnosis. We adapted reverse transcriptase polymerase chain reaction (RT-PCR) to amplify thyroid-typical mRNAs in samples of thyroid carcinoma cells as small as those obtained by fine-needle aspiration biopsy (FNAB), ie, 100-1000 cells, and applied this procedure to four routinely taken FNABs. Gene products such as thyroglobulin (Tg), thyroid stimulating hormone-receptor (TSHr), sodium/iodide-symporter (NIS), type I iodothyronine-5'-deiodinase (DI), and type II iodothyronine-5'-deiodinase (DII) were analyzed. To establish RT-PCR protocols, serial dilutions of follicular thyroid carcinoma cells, FTC-133, which express these genes at low levels, were initially used for RNA isolation. Successful RNA isolation and reverse transcription were checked by the amplification of beta-actin mRNA. We detected the mRNAs coding for Tg in as little as 10 cells, for NIS in 100 cells, and for TSHr, DI, and DII in 10,000 cells. After preparing cytological smears of four routinely taken FNABs, all above-mentioned thyroid-typical mRNAs were observed by using the material remaining in the needle for RNA isolation followed by RT-PCR. This method offers the possibility of obtaining two different types of information from the same routinely taken thyroid FNAB: the cytological diagnosis and the expression pattern of several diagnostically relevant genes. Therefore, a more specific diagnosis could be rendered in the preoperative state, and may lead to more specific therapy. PMID- 9848711 TI - Role of ultrasound-guided fine-needle aspiration biopsy in evaluation of nonpalpable thyroid nodules. AB - To determine the role of ultrasound-guided fine-needle aspiration biopsy (US FNAB) in the evaluation of incidentally diagnosed nonpalpable thyroid nodules (NP TN), we compared the diagnostic yield of US-FNAB in NP-TN larger than 10 mm in diameter, with palpation-guided (P-FNAB) in palpable thyroid nodules (P-TN) in the same hospital setting. Of 108 consecutive patients with NP-TN (female/male: 97/11, age: 55.2 +/- 14.3, mean +/- SD), malignancy was identified in 8 patients (7 with thyroid papillary carcinoma, 4 of which had extrathyroidal spread, and 1 patient had metastasis of pulmonary adenocarcinoma). Male gender was associated with higher prevalence of malignancy (p = 0.09) while calcification and/or cystic degeneration of the nodule did not predict the cytological findings. Malignancy was identified in 16 of 151 consecutive patients with P-TN (female/male: 139/12, age: 44.1 +/- 9.5, mean +/- SD) indicating a similar rate of malignancy in both NP-TN and P-TN. In the P-TN group thyroid cancer was more common in males (p = 0.007), and the US properties of the nodule did not predict its cytological diagnosis. In conclusion, because similar rates of cancer are detected by FNAB in both NP-TN and P-TN, an FNAB is recommended for NP-TN larger than 10 mm. PMID- 9848712 TI - The extracellular thyrotropin receptor domain is not a major candidate for mutations in toxic thyroid nodules. AB - Constitutive activation of the cyclic adenosine monophosphate (cAMP) cascade by either thyrotropin receptor (TSHR) or gsp mutations is considered to be the major molecular cause of toxic thyroid nodules (TTNs). In a recent study we investigated a consecutive series of 31 TTNs and identified 15 somatic TSHR mutations (n = 14 in exon 10; n = 1 in exon 9) but no mutations in gsp exons 7 10. The purpose of the present study was to determine whether the extracellular TSHR domain would be a candidate for mutations causing TTNs. Therefore, we screened TSHR exons 1-8 in the remaining 16 TTNs without mutations in TSHR exons 9 and 10 and gsp exons 7-10 of our previous study. Except for a known functional polymorphism in exon 1 (Pro 52 Thr) in 2 TTNs and a silent base exchange in exon 7 (187 Asn) in 7 other TTNs no TSHR mutations were identified. To clarify the molecular etiology of TTNs without TSHR or gsp mutations, candidate genes in other steps of the cAMP cascade have to be considered. PMID- 9848713 TI - Low prevalence of the ret/PTC3r1 rearrangement in a series of papillary thyroid carcinomas presenting in Belarus ten years post-Chernobyl. AB - After the Chernobyl accident in 1986, there was a significant increase in the incidence of papillary thyroid cancer in fallout-exposed children from Belarus. Radiation-induced rearrangements of chromosome 10 involving the c-ret proto oncogene have been implicated in the pathogenesis of these cancers. The ret/PTC3r1 rearrangement was the most prevalent molecular lesion identified in post-Chernobyl papillary thyroid cancers arising in 1991 and 1992. We identified the ret/PTC1 rearrangement in 29% of 31 papillary thyroid cancers presenting in Belarus in 1996. In the present report, we examined 14 cases from this series (plus 1 additional case) and found a ret/PTC3r1 rearrangement in only 1 (7%). The prevalence of ret/PTC3r1 in this series is significantly lower than previously reported (p = 0.0006, Fisher exact test). This result suggests a switch in the ratio of ret/PTC3 to ret/PTC1 rearrangements in late (1996) versus early (1991 1992) post-Chernobyl papillary thyroid cancers. PMID- 9848714 TI - Conception after iodine-131 therapy for differentiated thyroid cancer. AB - We reviewed the medical history of 26 women with differentiated thyroid cancer who became pregnant after receiving therapeutic doses of radioactive iodine (131I) in order to document the possibility of a greater risk for disorders in these pregnancies. There were a total of 39 pregnancies, 6 of which occurred during the first year after therapy. In 3 cases the following anomalies were encountered: a male suffering Trisomy 18 (Edward's syndrome), a female with constitutional aplastic anemia, and a male with a congenital hip dysplasia. Of the 33 pregnancies that occurred after the first year post-therapy, there were 2 spontaneous abortions and a male effected by ureteral stenosis. Based on these data, although it cannot be confirmed that these congenital disorders are due to the 131I therapy, we recommend that pregnancy be avoided for the first year after therapy. PMID- 9848715 TI - Interferon-gamma gene microsatellite polymorphisms in patients with Graves' disease. AB - Although some of the susceptibility to Graves' disease is conferred by genes in the human leucocyte antigen (HLA) region on the short arm of chromosome 6, other genetic factors must also predispose. Among the cytokines involved in thyroid autoimmunity interferon-gamma (IFN-gamma) plays a key role in the pathogenesis of Graves' disease. We therefore analyzed the first intron of the IFN-gamma gene for a dinucleotide (CA) repeat polymorphism on chromosome 12q. Two hundred two Caucasian patients with Graves' disease and 214 Caucasian controls were analyzed by polymerase chain reaction (PCR) and subsequent polyacrylamide gel electrophoresis technique: eight different alleles designated as IFN-gamma*1 to IFN-gamma*8 could be differentiated. Among Graves' disease patients IFN-gamma*5 (12.9% vs. 6.8%, p < 0.04) was significantly more frequent whereas IFN-gamma*2 (2.5% vs. 9.8%, p < 0.002) was significantly less frequent. Patients positive for the genetic susceptibility marker HLA DQA1*0501 had significantly more IFN gamma*3 alleles (13.6% vs. 2.6%, p < 0.009) and IFN-gamma*5 alleles (22.1% vs. 7.6%, p < 0.03) compared with DQA1*0501 positive controls. Also, among patients with endocrine ophthalmopathy IFN-gamma*3 (17.9% vs. 4.2%, p < 8 x 10(-6)) and IFN-gamma*5 (18.9% vs. 7.0%, p < 0.003) were significantly more frequent compared with controls. Although a significant association of IFN-gamma microsatellite polymorphism was observed, only a small proportion of Graves' disease patients have these markers. Thus, it is likely that the detected microsatellite polymorphisms play only a minor role in the susceptibility to Graves' disease. PMID- 9848716 TI - Natural killer cell activity in patients with Graves' disease and Hashimoto's thyroiditis. AB - Natural killer (NK) cell activity of peripheral blood lymphocytes (PBL) against k562 human tumor cell targets was studied in patients with Graves' disease and Hashimoto's thyroiditis. NK activity was measured in a standard 4-hour 51chromium (Cr) release assay. Cytotoxicity was expressed as lytic units (LU)/10(6) PBL. Significantly decreased NK cell activity was demonstrated in both groups of patients, with mean (+/- SE) lytic units of 10.3 (+/- 9.1) and 13.3 (+/- 10.3) for patients with Graves' disease and Hashimoto's thyroiditis, respectively, compared with 36.0 (+/- 26.3) for age- and sex-matched normal subjects. When patients with Graves' disease were analyzed according to their thyroid status; NK activity was significantly depressed in (1) hyperthyroid patients before treatment; (2) hyperthyroid patients receiving antithyroid therapy; and (3) euthyroid patients receiving antithyroid therapy, compared with normal subjects. Graves' disease patients who were hypothyroid after radioactive iodine therapy or thyroidectomy had normal NK activity. No significant differences between hyperthyroid and euthyroid patients or between hypothyroid patients and normal subjects were demonstrated. NK activity in patients with Graves' disease did not correlate with serum levels of thyroxine, the presence or severity of ophthalmopathy, or titers of serum thyroid antibodies. In patients with Hashimoto's thyroiditis there was no correlation between NK activity and goiter size, titers of antithyroid antibodies, or thyroid status. These findings suggest that depression of NK activity in both disorders is secondary to abnormalities of thyroid hormone secretion, although an effect of the underlying autoimmune reactions has not been excluded. PMID- 9848717 TI - Thyroid-associated ophthalmopathy in black South African patients with Graves' disease: relationship to antiflavoprotein antibodies. AB - Thyroid-associated ophthalmopathy (TAO) is a progressive eye disorder associated with Graves' hyperthyroidism, which is generally considered to have an autoimmune etiology. Eye muscle membrane proteins of 64 kd are good markers of ophthalmopathy in patients with thyroid autoimmunity. The 64-kd protein is now shown from a partial sequence to be the flavoprotein subunit (Fp) of mitochondrial succinate dehydrogenase. Hyperthyroidism due to Graves' disease is increasing in incidence among urban black female Africans, possibly because of exposure to environmental risk factors such as increased dietary iodine ingestion and stress. Ophthalmopathy is frequently observed in this clinical context, but its association with serum autoantibodies reactive with Fp has not been examined. We studied 19 black South African patients with Graves' disease during the course of prolonged antithyroid drug administration, of whom 10 had congestive ophthalmopathy, but no clinical evidence for eye muscle damage at the onset. Anti Fp antibodies were detected in 2 of these patients, as well as in 2 of the 9 patients who did not have overt eye disease. Additionally, the antibodies became positive in 3 patients with ophthalmopathy in whom tests were negative initially, remained positive in 1 patient throughout the study period and became negative in 1 patient with positive tests initially. Ophthalmopathy did not develop in any of the 9 patients who lacked this complication on presentation. The reasons why we failed to demonstrate a close relationship between anti-Fp antibodies and the eye muscle component of ophthalmopathy are unclear although one possibility is that ocular myopathy is an uncommon manifestation in African thyrotoxic patients compared with those of Caucasian origin. The relationship between anti-Fp antibodies and eye muscle inflammation in patients with thyroid autoimmunity of different ethnic origins and environmental settings, needs to be addressed in a large prospective study. PMID- 9848718 TI - Serum creatine kinase levels in overt and subclinical hypothyroidism. AB - Overt hypothyroidism is known to be associated with increased serum creatine kinase (CK) levels. However, there is little information on CK levels in subclinical hypothyroidism. The aim of the study was to assess the relationship between CK levels and thyroid function in overt and subclinical hypothyroidism. Thyroid function tests (thyrotropin [TSH], free thyroxine [FT4], free triiodothyronine [FT3]) and the serum levels of CK were obtained from 23 patients admitted to a general hospital for illnesses other than thyroid or muscular diseases, myocardial ischemia, or brain damage. Overt hypothyroidism, based on thyroid function tests, was present in 10 patients, whereas hypothyroidism could be classified as subclinical in the other 13. A positive correlation was observed between CK and thyrotropin, and to a lesser extent between CK and thyroid hormones. Moreover, the correlation between CK and TSH and between CK and FT4 was detectable in subclinical hypothyroidism. Our data suggest that even in subclinical hypothyroidism there is some degree of dysfunction in skeletal muscle metabolism. PMID- 9848719 TI - Myopathy as the persistently isolated symptomatology of primary autoimmune hypothyroidism. AB - Although disorders of thyroid function may cause a wide range of muscle disturbances, an overt myopathy has been rarely reported as an isolated clinical presentation of hypothyroidism. We observed 10 patients (5 males and 5 females) who had been referred to the department of neurology because of muscular fatigability, myalgia, cramps, or proximal weakness. Laboratory investigation showed that all patients had hypothyroidism due to Hashimoto's thyroiditis (atrophic variant in 9/10). Classic symptoms/signs of hypothyroidism such as lethargy, constipation, cold intolerance, myxedematous facies, and/or bradycardia were absent, as assessed independently by the three coauthoring thyroidologists. Muscular complaints improved greatly and then disappeared after substitutive levothyroxine treatment. Muscle biopsy revealed nonspecific changes. Nicotinamide adenine dinucleotide reductase (NADH-TR)-hyporeactive cores were present in two patients (10% and 90% of type 1 fibers). On electron microscopy, the core areas showed disorganized myofibrils, Z-band streaming, rod formation, and paucity of mitochondria and glycogen granules. Desmin intermediate filaments were overexpressed only in some cores. The similarity of the pattern of desmin expression between hypothyroid cores and target lesions of denervated fibers supports the hypothesis that, at least in some of our patients, myopathy was the result of an impaired nerve-mediated action of thyroid hormones on skeletal muscle. Our observations suggest that an isolated myopathy as the sole manifestation of hypothyroidism is not a rare event. We postulate that our cases may constitute a peculiar subgroup of Hashimoto's thyroiditis patients: (1) the strikingly abnormal F/M ratio of 1:1; (2) the relatively younger age; (3) the rarity of the goitrous variant; (4) the unusual finding of antithyroglobulin (Tg Ab) > antithyroid peroxidase (TPO-Ab). Thorough evaluation of thyroid function is appropriate in patients with myopathy of uncertain origin. PMID- 9848720 TI - Sporadic medullary microcarcinoma of the thyroid: a retrospective analysis of eighty cases. AB - Clinical characteristics and prognosis of 80 patients (53 women and 27 men) with sporadic medullary thyroid carcinomas (MTC), less than 1 cm in size (micro-MTC), operated on between 1971 and 1996 are reported (73 total and 7 partial thyroidectomies). These patients, obtained from a national database of 899 patients with MTC, were compared with 357 cases of sporadic MTC greater than 1 cm and 149 subjects with familial MTC less than 1 cm (familial micro-MTC). Median age at surgery was 52.5 years, a distribution similar to larger sporadic MTC. Micro-MTC was identified due to elevated calcitonin (47.5%), clinically identified lymph node (10.0%), distant metastases (6.3%) or pathologic finding at surgery (36.2%). Diarrhea and/or flushing were observed in 6 patients including 4 with clinically identified lymph node. Among patients who had lymph node dissection at surgery (68.8%), lymph node involvement with tumor was observed in 30.9%, and was significantly more frequent in multifocal (7/11) than in unifocal micro-MTC (p < 0.03). All sporadic micro-MTC were unilateral. Survival rate was 93.9% +/- 4.4% (SE) at 10 years, greater than that observed in sporadic macro-MTC (p = 0.04). Normal postoperative basal calcitonin (CT) was obtained in 71.1% of micro-MTC patients versus 33.6% in sporadic macro-MTC (p < 0.01). Sporadic micro MTC is much more frequent than expected, 15% of MTC in our series. Although specific survival rate and percentage of biological cure in micro-MTC are significantly better than for larger tumors, the frequency of lymph node involvement, however, justifies an aggressive surgical approach including total thyroidectomy and bilateral central lymph node dissection. PMID- 9848721 TI - Papillary and follicular thyroid carcinoma metastatic to the skin: a case report and review of the literature. AB - Cutaneous metastases from thyroid cancers are rare. We report the case of an otherwise asymptomatic 81-year-old woman with an enlarging scalp lesion. Her solitary skin metastasis was the presenting feature of thyroid carcinoma. Routine histopathology of the lesion was notable for an atypical clear cell neoplasm. Immunohistochemistry was positive for thyroglobulin. Subsequent resection of the thyroid gland identified separate foci (< 1 cm) for both papillary and follicular carcinoma. Although such immunohistochemical staining has been used previously, it has never been reported to provide the definitive diagnosis for a solitary cutaneous metastasis from the thyroid. Previous tumors had anatomic features in a clinical context that permitted identification by routine light microscopy. Clear cell features found in the follicular focus of carcinoma in the thyroid suggest that it is the primary. A worldwide literature review reveals that follicular carcinoma has a greater preponderance than papillary carcinoma for cutaneous metastasis and that the majority of skin metastases from either papillary or follicular thyroid cancer are localized to the head and neck. PMID- 9848722 TI - Amyloid goiter due to primary systemic amyloidosis: a diagnostic challenge. AB - We describe a euthyroid patient who presented with a goiter that continued to enlarge despite levothyroxine administration. Three fine-needle aspirations for cytology were nondiagnostic. An open biopsy was complicated by bleeding from the surgical site. Primary systemic amyloidosis was diagnosed on the basis of the goiter histology, bone marrow aspirate, and urine immunoelectrophoresis. The patient received melphalan and steroid treatment and survived for an additional 16 months. This period was complicated by congestive heart failure, generalized seizures, and upper gastrointestinal bleeding. Our case illustrates the difficulties in making the diagnosis and in treatment of primary systemic amyloidosis. PMID- 9848723 TI - Lingual thyroid: report of three cases. AB - Lingual thyroid is a rare developmental disorder and is more frequent in women. The pathogenesis is unclear but may be related to the presence of maternal blocking autoantibodies against the thyroid. Treatment of this disorder includes the use of levothyroxine in order to correct the hypothyroidism, which is very frequent and to induce the shrinkage of the gland. When symptoms of obstruction or bleeding appear, ablative therapy by means of surgery or radioiodine is warranted. We report three cases and discuss the approach to diagnosis and a strategy for management. PMID- 9848724 TI - Expression of thyrotropin receptor on clonal osteoblast-like rat osteosarcoma cells. AB - The expression of thyrotropin receptor (TSH-R) on various cells derived from bone, including osteoblast-like rat osteosarcoma cells (UMR106 cells), was investigated. TSH receptor mRNA was detected in UMR106 cells by Northern blot analysis. 125I-labeled TSH binding analysis revealed specific high- and low affinity binding sites (association constants of 5.6 x 10(9) M(-1) and 3.0 x 10(7) M(-1), respectively) on UMR106 cells. Recombinant TSH, but not recombinant human chorionic gonadotropin, increased cyclic adenosine monophosphate (cAMP) production in a concentration-dependent manner in these cells. Furthermore, immunoglobulin Gs from patients with Graves' disease induced cAMP response in UMR106 cells, and the cAMP response index in this cell line correlated with thyroid-stimulating antibody (TSAb) activity detected by Chinese hamster ovary (CHO)-K1 cells transfected with rat TSH-R. We have also demonstrated that recombinant TSH increased cAMP production in human osteoblast-like osteosarcoma (MG63) cells and mouse primary osteoblastic cells. These results suggest that osteoblasts possess functional TSH-R and that abnormal bone metabolism in Graves' disease may be partly explained by the interaction of TSAb with TSH-R in osteoblasts in some patients. PMID- 9848725 TI - Transforming growth factor-beta1 resistance in a thyroid cancer model of tumor necrosis factor-alpha resistance. AB - We have shown that both tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) inhibit the growth of the human papillary thyroid carcinoma (PTC) cell line, NPA. In previous work, we developed NPA cells that were resistant to the growth suppressive effect of TNF-alpha, called R30, R45, and R60. In this model there were alterations in the p55 and p75 TNF-alpha receptor signaling in the resistant cell lines. In the present work, we studied the action of TGF-beta1 in this PTC cell model. TGF-beta1 (111 pg/mL) inhibited the proliferation of NPA, R30, R45, and the R60 cell lines by 82.8%, 72.1%, 64.2%, and 24.2%, respectively. On Western analysis, TGF-beta1 reduced c-fos content with similar potency in the NPA and R60 cells. In contrast, TNF-alpha reduced c-fos content in the sensitive NPA cells, but failed to do so in the resistant R60 cells. TGF-beta1 reduced p53 content in the NPA but not in the R60 cells, while TNF-alpha did not affect the p53 content in these cells. Furthermore, the resistant cells had a lower baseline p53 content than the NPA cells. The resistant cells had a significantly increased growth rate. Enzyme linked immunosorbent assay (ELISA) assays with specific antibody against human p53 showed no apparent increase in the mutant form of p53 in the resistant cells. There were also no mutant forms of Ha-Ras, Arg12p21, Val12p21, Asp12p21, and Asp13p21 detected in the resistant cells. The results showed that R30, R45, and R60 cells are partially resistant to TGFbeta1. The mechanisms of action of TNF alpha and TGF-beta1 differ in their regulation of c-fos and p53 content. The increase in cell proliferation rate is apparently associated with a decrease of p53 content, but not with mutations of p53 or Ha-Ras. PMID- 9848726 TI - The neuropeptides, VIP and NPY, that are present in the thyroid nerves are not released into the thyroid vein. AB - In the present study, we tested the hypothesis that the neuropeptides, vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY), which are present in the thyroid nerves, act as physiological neurotransmitters involved in the regulation of thyroid hormone secretion and thyroid blood flow. Specifically, we examined whether these neuropeptides can be released into thyroid blood vessels by electrical stimulation of the major thyroid nerves or whether their expression is altered by changes in iodine intake. Sprague-Dawley rats were used in this study. The cervical sympathetic trunk or the superior laryngeal nerve was stimulated by bipolar electrodes in anesthetized rats. During nerve stimulation, blood samples were withdrawn from the thyroid vein. Thyroid blood flow was monitored by laser Doppler blood flowmetry. Sympathetic stimulation caused a marked decrease in thyroid blood flow, which was associated with a significant increase in release of norepinephrine. However, these effects were not accompanied by any change in NPY release into the thyroid vein. Stimulation of the superior laryngeal nerve was not associated with changes in thyroid blood flow or VIP release into the thyroid vein. In a separate experiment, rats were fed a diet containing low-, high-, or normal iodine concentrations. Triiodothyronine (T3) and thyroxine (T4) levels in thyroid venous plasma were significantly reduced in rats fed a low iodine diet but not in a separate group of rats fed a high iodine diet. However, these treatments had no effect on VIP or NPY concentrations in thyroid venous plasma or in thyroid ganglia. Thus, our results indicate that VIP and NPY, which are present in the thyroid nerves, may not be directly involved in the regulation of thyroid function. PMID- 9848727 TI - In vivo 1H MRS of normal breast and breast tumors using a dedicated double breast coil. AB - Image-guided localized proton MR spectroscopy (MRS) of normal breasts and breast tumors (ductal and undifferentiated carcinomas) was performed using a dedicated double breast coil. In vivo 1H MR spectra from 10 normal volunteers showed signals from water and lipids only, even in breasts with small contribution of fatty breast tissue. In the spectra from 6 of the 12 examined patients, an intense signal assigned to choline compounds was detected. The signal was also detected at lower levels in the remaining patients. This study shows that in vivo 1H MRI/MRS examinations of breast tumors can be performed within an examination time of 45 to 60 minutes. Signals from breast tumor metabolites may be detected using in vivo 1H MRS. PMID- 9848728 TI - Cystic adenomyosis of the uterus: MRI. AB - Adenomyosis of the uterus is most often seen as focal or diffuse thickening of the myometrial junctional zone on MRI. We describe the morphologic features and signal characteristics of the rarer cystic form of the disease, as revealed by MRI. We conclude that cystic adenomyosis of the uterus is characterized by a well circumscribed cystic lesion within the myometrium that demonstrates hemorrhage in differential stages of organization on MR images. PMID- 9848729 TI - Hyperreactio luteinalis in a normal pregnancy: sonographic and MRI findings. AB - A case of hyperreactio luteinalis in an otherwise normal pregnancy is reported. Ascites was present, but no peritoneal implants or adenopathy were seen. Findings that would have suggested the correct diagnosis are the symmetrical and bilateral pattern of the mass, as well as the rather uniform size of the loculi, which were 1 to 3 cm in diameter. PMID- 9848730 TI - Comparison of ultrafast half-Fourier single-shot turbo spin-echo sequence with turbo spin-echo sequences for T2-weighted imaging of the female pelvis. AB - So that we might evaluate the ultrafast half-Fourier single-shot turbo spin-echo (HASTE) sequence in T2-weighted MRI of the female pelvis and compare it with the turbo spin-echo (TSE) sequence, we prospectively studied 60 consecutive females with suspected abnormalities of the pelvis. For all MR examinations, we used a 1.5-T superconductive magnet with a phased array coil. The HASTE sequence was applied with TR/effective TE/echo train = infinity/90/64 and a 128 x 256 matrix (acquisition time: .3 sec/slice), conventional TSE imaging with 3,400 to 5,000/132/15 and a 128 x 256 matrix (mean acquisition time: 2 min 4 sec), and high-resolution TSE imaging with 3,400 to 5,000/132/15 and a 300 x 512 matrix (6 min 4 sec). Although the lesion conspicuity for the HASTE sequence was less than that for the high-resolution TSE sequences, artifacts (including ghosting, bowel motion, susceptibility difference, and chemical shift) were negligible on HASTE images of all patients. The lesion conspicuity for the HASTE sequence was significantly better than for the conventional TSE sequence. In spite of the very short acquisition time, the subjective scoring of the overall image quality for the HASTE sequence was significantly higher than for the conventional TSE sequence (P < .01) and were slightly lower than for the high-resolution TSE sequence. Compared with high-resolution TSE, HASTE provided clearer visualization of large leiomyomas and ovarian tumors but slightly poorer visualization of uterine cancer. In occlusion, HASTE sequence generates higher contrast and is free from motion and chemical shift artifact with much higher time efficacy. Because of limited image resolution, the HASTE sequence should be used when the high-resolution TSE imaging is suboptimal. PMID- 9848731 TI - T2-weighted breath-hold MRI of the liver at 1.0 T: comparison of turbo spin-echo and HASTE sequences with and without fat suppression. AB - To compare the clinical usefulness of T2-weighted breath-hold sequences for imaging the liver, 33 patients with 97 focal hepatic lesions were studied with a 1.0-T scanner by using T2-weighted breath-hold turbo spin-echo (SE) sequences and T2-weighted breath-hold half-Fourier single-shot turbo SE (HASTE) sequences with and without fat suppression. Images were quantitatively analyzed for liver signal to-noise ratio (SNR) and lesion-to-liver contrast-to-noise ratios (CNR). Qualitative analysis was performed for lesion conspicuity, motion artifacts, and anatomic sharpness of extrahepatic structures. Breath-hold turbo SE imaging with fat suppression showed the highest CNR for cystic lesions and the best lesion conspicuity for cystic and solid lesions among the four sequences. For solid lesions, there was no significant difference of lesion-to-liver CNR between them. HASTE sequence was superior to turbo SE sequences in terms of motion artifacts; however, the usefulness for evaluating focal hepatic lesions was limited compared with turbo SE sequence with fat suppression. Addition of fat suppression was not helpful for HASTE imaging because of decreased lesion conspicuity and extrahepatic details without the advantage of reducing motion artifacts. This study suggests that turbo SE sequence with fat suppression is most useful for breath-hold T2-weighted liver imaging at 1.0 T. Addition of imaging without fat suppression can be considered for evaluating extrahepatic structures. HASTE sequence may have a role for imaging uncooperative patients due to absence of motion artifacts. PMID- 9848732 TI - Fast and ultrafast magnetic resonance imaging in renal lesions. AB - Our purpose was to analyze and compare the image quality and contrast-to-noise ratio (CNR) of different fast T1- and T2-weighted sequences with conventional spin-echo sequences in renal MRI. Twenty-three patients with focal renal lesions were examined with a T2-weighted ultrafast turbo spin-echo (UTSE) sequence with and without frequency selective fat suppression (SPIR), a combined gradient-and spin-echo sequence (GraSE), and a conventional spin-echo sequence (SE). In addition, T1-weighted images were obtained pre- and postcontrast, using a fast spin-echo sequence (TSE) with and without SPIR and the conventional SE sequence. Among the T2-weighted images, the highest CNR and the best image quality were obtained with the UTSE sequence, followed by the fat-suppressed UTSE sequence. GraSE and conventional SE sequences showed a significantly lower CNR and image quality (p < 0.05). The T1-weighted sequences did not show significant differences, in either precontrast or postcontrast measurements. T2-weighted UTSE with and without fat suppression combined excellent image quality and high CNR for imaging and detection of renal lesions. The T1-weighted fast sequences provided no alternative to the gradient-echo or to the conventional SE sequences. The results of this systematic study suggest the use of T2-weighted fast techniques for improved diagnostic accuracy of renal MRI. PMID- 9848733 TI - Limits to the accuracy of vessel diameter measurement in MR angiography. AB - This work addresses the fundamental limits imposed by the MRI process on the accuracy with which vessel diameters and cross-sectional areas can be derived from time-of-flight (TOF) and phase-contrast (PC) MR source images. By means of simulations and in vitro experiments, it is demonstrated that, even in the absence of flow-related artifacts, severe inaccuracies in the determination of diameters or cross-sectional areas may occur solely because of the physical process of the MR image acquisition. Resolution and intraluminal saturation have strong effects on the vessel appearance and thus on the diameter estimation error. It is shown that low resolution leads to diameter overestimation or even underestimation and that intraluminal saturation causes severe underestimation, even for relatively low flip angles. Velocity and velocity encoding do not have a major influence on lumen appearance in PC images. Accurate diameter estimations can be attained only if lumen diameters constitute at least three pixels for both TOF and PC acquisitions, provided that intraluminal saturation is suppressed or avoided. Additionally, since the constitution of TOF and PC images is dissimilar, lumina should be analyzed differently to obtain accurate diameters and cross sectional areas. PMID- 9848734 TI - In vivo correlation between blood T2* and oxygen saturation. AB - The in vivo relationship between blood T2* and oxygen saturation was investigated. A wide range of blood oxygen saturation levels was created in pigs by altering the ventilation rate. Blood T2* was measured in vivo in the abdominal aorta and the inferior vena cava, corresponding to each oxygen saturation level. Our results indicate that it is possible to measure blood T2* in vivo reliably with an MR technique we previously proposed. Blood T2* correlates closely with oxygen saturation levels over a wide range, and the relationship between blood T2* measured in vivo and oxygen saturation can be approximated by a linear function. PMID- 9848736 TI - MRI of patellar articular cartilage: evaluation of an optimized gradient echo sequence (3D-DESS). AB - Our purpose was to evaluate the diagnostic efficacy of a gradient-echo sequence optimized for cartilage imaging in patellar cartilage abnormalities and to compare it to a standard turbo-spin-echo sequence. Fifty-eight consecutive patients who underwent, within 3 months both MRI and arthroscopy or surgery, were included in the investigation. Two radiologists specializing in musculoskeletal imaging independently assessed axial three-dimensional double-echo steady state (3D-DESS) gradient-echo images and sagittal proton- and T2-weighted turbo-spin echo images with regard to retropatellar cartilage abnormalities. Possible findings were: 0: normal, 1: cartilage softening, and 2: lesion of the articular surface. Inter- and intraobserver variability was assessed. For cartilage softening, the axial 3D-DESS sequence had a sensitivity of 73%, a specificity of 75%, and an accuracy of 70%. The corresponding results for the sagittal turbo spin-echo sequence were 53%, 65%, and 62%. For surface lesions, the results for the 3D-DESS sequence were 43%, 92%, and 83% and for the turbo-spin-echo sequence were 60%, 92%, and 86%. Intra- and interobserver agreement was moderate (k = 0.59 and 0.45 [DESS], 0.6 and 0.46 [turbo -spin-echo]). We conclude that the 3D-DESS sequence is moderately accurate in detecting patellar cartilage abnormalities. Compared with the sagittal turbo-spin-echo sequence, the axial 3D-DESS sequence is superior in diagnosing cartilage softening but not surface lesions. PMID- 9848735 TI - In vivo quantitative mapping of cardiac perfusion in rats using a noninvasive MR spin-labeling method. AB - Measurement of myocardial perfusion is important for the functional assessment of heart in vivo. Our approach is based on the modification of the longitudinal relaxation time T1 induced by magnetic spin labeling of endogenous water protons. Labeling is performed by selectively inverting the magnetization within the detection slice, and longitudinal relaxation is measured using a fast gradient echo MRI technique. As a result of blood flow, nonexcited spins enter the detection slice, which leads to an acceleration of the relaxation rate. Incorporating this phenomenon in a mathematical model that describes tissue as two compartments yields a simple expression that allows the quantification of perfusion from a slice-selective and a global inversion recovery experiment. This model takes into account the difference between T1 in blood and T1 in tissue. Our purpose was to evaluate the feasibility and reproducibility of this technique to map quantitatively myocardial perfusion in vivo in rats. Quantitative maps of myocardial blood flow were obtained from nine rats, and the reproducibility of the technique was evaluated by repeating the whole perfusion experiment four times. Evaluation of regions of interest within the myocardium yielded a mean perfusion value of 3.6 +/- .5 ml x min(-1) x g(-1) over all animals, which is in good agreement with previously reported literature values. PMID- 9848737 TI - Vital sign changes during infant magnetic resonance examinations. AB - Heart rate (HR) and blood oxygen saturation (SaO2) were monitored before and during clinically indicated MR examinations of newborns to (a) identify any temporal relationship between MR scanning and vital sign fluctuations and (b) assess the reliability of SaO2 monitoring of dynamic changes. Fluctuations in HR (but not in SaO2) that are temporally linked to the MR image acquisition occur in most neonates during routine clinical MR examinations. PMID- 9848738 TI - Magnetic resonance imaging of shear wave propagation in excised tissue. AB - The propagation of shear waves in ex vivo tissue samples, agar/gel phantoms, and human volunteers was investigated. A moving coil apparatus was constructed to generate low acoustic frequency shear perturbations of 50 to 400 Hz. Oscillating gradients phase-locked with the shear stimulus were used to generate a series of phase contrast images of the shear waves at different time-points throughout the wave cycle. Quantitative measurements of wave velocity and attenuation were obtained to evaluate the effects of temperature, frequency, and tissue anisotropy. Results of these experiments demonstrate significant variation in shear wave behavior with tissue type, whereas frequency and anisotropic behavior was mixed. Temperature-dependent behavior related mainly to the presence of fat. Propagation velocities ranged from 1 to 5 m/sec, and attenuation coefficients of from 1 to 3 nepers/unit wavelength, depending on tissue type. These results confirm the potential of elastic imaging attributable to the intrinsic variability of elastic properties observed in normal tissue, although some difficulty may be experienced in clinical implementation because of viscous attenuation in fat. PMID- 9848739 TI - Determination of focal ischemic lesion volume in the rat brain using multispectral analysis. AB - Multispectral (MS) analysis was used to determine the ischemic lesion volume in the rat brain after permanent middle cerebral arterial occlusion. MS analysis used a four-dimensional MS model consisting of an estimate of the average apparent diffusion coefficient of water (ADC(av)), T2, proton density, and perfusion. Four classification methods were investigated: (a) multivariate gaussian (MVG); (b) k-nearest neighbor (k-NN); (c) k-means (KM); and (d) fuzzy c means (FCM). MVG and k-NN classifiers are supervised methods requiring labeled training data to characterize the stroke lesion. Unsupervised classifiers (KM, FCM) do not require previous statistics or labeled training data, resulting in potentially greater clinical usefulness. All MS methods provided significant correlation with postmortem findings beyond the use of ADC(av) alone (partial correlation given the ADC(av) estimate: MVG, .66; k-NN, .75; KM, .68; FCM, .70). This study demonstrates that MS analysis provides an improved estimate of ischemic lesion volume over that obtained from ADC alone. PMID- 9848740 TI - Fat suppressed MRI of articular cartilage with a spatial-spectral excitation pulse. AB - We developed a three-dimensional, gradient-recalled-echo imaging technique that incorporates a short-duration spatial-spectral excitation pulse from the family of binomial pulses. Binomial pulses of different orders were tested on phantoms and on normal volunteers to find the composite pulse that produced in the shortest duration the most reliable fat suppression. Composite pulses employing unipolar slice-selective gradients with explicit rewinder gradients between each radio-frequency (RF) pulse were compared with composite RF pulses employing alternating-polarity, slice-select gradients. The advantage of the sequences using the unipolar gradients is improved fat suppression. Images of the knees of volunteers produced with the composite RF pulse have contrast between fat and articular cartilage equivalent to that on images created by the gradient-recalled echo imaging technique employing a conventional chemsat pulse. The optimum RF pulse consisted of three amplitude- and phase-modulated pulses combined with unipolar slice-select gradients. PMID- 9848741 TI - Locally focused MRI of interventions. AB - Certain interventional MR procedures would benefit from T2-weighted imaging because of the sensitivity of T2-weighted images to tissue damage and target lesion contrast. To acquire such images with reasonable temporal resolution, a single-shot acquisition should be used because of the inherently long TR needed for T2 weighting. Unfortunately, most scanners require long readout times (eg, greater than 150 msec) and high bandwidths (eg, greater than 120 kHz) to perform conventional single-shot imaging with high spatial resolution. The resulting images are thus degraded by unacceptable artifacts and noise levels. This study illustrates how to create locally focused MR images that have high spatial resolution in a region of interest and lower spatial resolution elsewhere in the image. Because these images can be created from sparse k-space data, a scanner with modest gradients (eg, 10 mT/m maximal amplitude, 500 microsec minimal rise time) can acquire them after a single excitation with relatively short readout time and low bandwidth. This technique may make it practical to monitor interventions with T2-weighted imaging. The method was illustrated by reconstructing dynamic changes, which were simulated experimentally by moving objects in the vicinity of a normal human head. PMID- 9848742 TI - Single-shot diffusion-weighted RARE sequence: application for temperature monitoring during hyperthermia session. AB - A diffusion-sensitive single-shot RARE (rapid acquisition with relaxation enhancement) sequence was implemented on a 2T whole-body MRI system. The sequence was optimized for diffusion-based MR thermometry, both on a conventional whole body gradient system and on a high-performance gradient insert. The use of spin echo versus stimulated-echo diffusion weighting is discussed as a function of gradient performance. Diffusion-based temperature mapping was used to observe the effect of the geometry of the antenna used for radiofrequency (RF) hyperthermia on the temperature distribution. Temperature changes of +/-.5 degrees C in gel and +/-2 degrees C in a muscle sample in vitro could be detected within 16 seconds (gel) or 1 minute (muscle) at a spatial resolution of 2 x 2 x 8 mm. Temperature changes in vivo were also observed on human muscle cooled with ice with comparable sensitivity for the measured apparent diffusion coefficient (ADC) values. PMID- 9848743 TI - MRI monitoring of laser ablation using optical flow. AB - The optical flow method is used for visualizing and quantifying the dynamics of tissue changes observed by MRI during thermal ablations. An approach was implemented for parallel two-dimensional optical flow calculations including the replacement of spurious velocities. Velocity magnitude results were found to be accurate in low-noise cases in tests using series of synthetic images. Optical flow results are presented from thermal ablation experiments utilizing a homogeneous polyacrylamide gel phantom and heterogeneous rabbit liver tissue in vivo, exhibiting heating and cooling with the accompanying quantitative characterization of the dilation and contraction of the thermally affected region. Results demonstrate that optical flow is capable of noninvasive real-time monitoring and control of interstitial laser therapy (ILT). PMID- 9848744 TI - Interactive MR-guided percutaneous nephrostomy. AB - Open-configuration MRI systems have been recently introduced and hold promise to allow the performance of a variety of minimally invasive procedures. Experience with MR-guided catheter-based luminal interventions is experimental to date. This is the first case of a successful percutaneous nephrostomy tube placement in a patient in an interventional MR system. The procedure was performed completely under MR guidance, and technical aspects are reviewed and compared with other, established techniques for percutaneous nephrostomy placement. PMID- 9848745 TI - Multiple pulmonary aneurysms secondary to Hughes-Stovin syndrome: demonstration by MR angiography. AB - We report a case of a 41-year-old patient with multiple pulmonary aneurysms secondary to Hughes-Stovin syndrome. Conventional pulmonary angiography could not be performed, because thrombus in both the inferior and superior vena cavae prevented catheter passage. Contrast-enhanced three-dimensional (3D) MR angiography (MRA) showed two segmental pulmonary aneurysms in the right lung. The affected segments of the lung were resected successfully, predicated on the MRA findings. PMID- 9848746 TI - Subungual glomus tumor: combined use of MRI and three-dimensional contrast MR angiography. AB - We describe a case of digital glomus tumor diagnosed by MRI and three-dimensional contrast MR angiography (MRA). Images provided the formal definitive diagnosis and the precise localization of the tumor, guiding the necessary surgical resection. It is possible that noninvasive MRA could replace conventional arteriography for the evaluation of patients with clinical suspicion of glomus tumor. PMID- 9848747 TI - Functional MRI of the lumbar spine in erect position in a superconducting open configuration MR system: preliminary results. AB - The purpose of this study was to determine the feasibility of obtaining, and findings in, functional MRI of the lumbar spine in an erect position and with flexion and extension. Thirty subjects (including 5 volunteers) were imaged in a sitting position and while performing flexion and extension. The alternations in posterior disk margin, size of neural foramina, and central canal were evaluated. In addition, routine supine imaging was accomplished in 15 of these subjects. The foraminal size and posterior disk margins did not change appreciably from supine to upright position. With extension, there was an increased disk bulge in 27% of disks (40% of those with desiccation). Central canal size (50%) and foraminal size (27%) decreased with extension, especially at levels with disk desiccation. Images obtained with our open-configuration MR unit were diagnostically adequate, although of inferior quality compared with those obtained with a conventional unit. Our preliminary results show the feasibility of obtaining diagnostic images of the erect lumbar spine with flexion and extension. The results are in agreement with those obtained with cadaveric studies. The utility of this method in diagnostic imaging of patients with low back pain remains to be determined. PMID- 9848749 TI - Cardiovascular catheters and accessories: ex vivo testing of ferromagnetism, heating, and artifacts associated with MRI. AB - The purpose of this study was to evaluate the MR safety of cardiovascular catheters and accessories. Intravascular cardiovascular catheters and accessories were tested for MR safety at 1.5 T using previously described techniques with respect to the evaluation of magnetic field attraction (deflection angle method), heating (temperature measured immediately before and after performing MRI), and artifacts (using a fast spoiled gradient-recalled acquisition in steady state [GRASS] pulse sequence). Two devices were attracted (RV pacing lead and Oximetrix 3 SO2 optical module) by the static magnetic field. Each of the other objects displayed no attraction. Heating was +0.2 degrees C for the sample cardiovascular catheter tested (Opticath). Artifacts varied from moderate to severe, depending on the amount and type of metal present in the device. Despite these ex vivo test results, further safety consideration should be given to the cardiovascular devices that have a conductive wire component (ie, certain types of the cardiovascular catheters) because of the potential for inducing current and excessive heating in these devices during MRI, especially using a high-field strength MR system. The cardiovascular catheters evaluated in this study or those with a similar design are not recommended for use in patients undergoing MRI procedures. PMID- 9848748 TI - Black-blood MR angiography with GRASE: measurement of flow-induced signal attenuation. AB - We investigated the feasibility of performing black-blood MR angiography (MRA) with the gradient and spin-echo (GRASE) pulse sequence. Phantom experiments and human testing were conducted, and the results were compared with those of turbo spin-echo (TSE). We demonstrated that both techniques are able to produce signal suppression of flowing fluid to background level. With fewer radiofrequency (RF) refocusing pulses, GRASE pulse sequences could serve as an alternative black blood technique of reduced RF power exposure and shorter scan time. These relative advantages of GRASE may become useful when high-resolution images are taken. PMID- 9848750 TI - A compact respiratory-triggering device for routine microimaging of laboratory mice. AB - A partial-body plethysmograph was developed for measuring the respiratory flow of anesthetized mice during routine microimaging experiments performed in the close confines of an 89-mm-diameter, vertical-bore magnet. Respiratory flow patterns were used for synchronizing conventional T2-weighted spin-echo imaging with the respiratory cycle, thereby, significantly reducing motion-induced artifacts and increasing observed liver lesion contrast. PMID- 9848751 TI - A phantom for diffusion-weighted imaging of acute stroke. AB - A tissue phantom for diffusion-weighted imaging was developed, basing its contrast between two compartments on different apparent diffusion coefficients, without contrast due to T2 relaxation and proton density. These contrast properties of the phantom simulate the situation found in normal gray matter and areas of acute ischemia. A possible application of the phantom was demonstrated for the investigation of the accuracy of volume measurements based on diffusion weighted images. PMID- 9848753 TI - California Association of Neonatologists: who, when, and why? PMID- 9848752 TI - Contrast-enhanced 3D MRA with centric ordering in k space: a preliminary clinical experience in imaging the abdominal aorta and renal and peripheral arterial vasculature. PMID- 9848754 TI - Sleep patterns in pregnancy: a longitudinal study of polysomnography recordings during pregnancy. AB - OBJECTIVE: To evaluate the differences in sleep of women throughout pregnancy compared with those of nonpregnant control subjects. STUDY DESIGN: Four pregnant women were studied longitudinally during their pregnancy using inpatient polysomnography. Measurements included electroencephalography, electrocardiography, and continuous-pulse oximetry. Four healthy nonpregnant women matched for age and weight were used as control subjects. The total sleep time was recorded, and percentages of each sleep stage were generated. RESULTS: Qualitative differences in sleep between pregnancy patients and control subjects were evident. Control subjects displayed a normal appearance of slow-wave sleep in stages 3 and 4 (delta sleep). When pregnant patients did display delta sleep, it appeared abnormal secondary to extensive alpha-wave intrusion. Even when including this abnormal delta sleep in a quantitative comparison, the pregnant patients had a significantly shorter percentage of total sleep time in delta sleep (4.9+/-1.9% vs 21.9+/-6.0%, p=0.03). CONCLUSION: Sleep in pregnancy is characterized by loss of normal slow-wave sleep. Thus, sleep stages 3 and 4 are shortened during pregnancy. This sleep alteration is persistent when followed longitudinally during pregnancy. PMID- 9848756 TI - Spontaneous minute ventilation predicts readiness for extubation in mechanically ventilated preterm infants. AB - OBJECTIVE: We designed an observational study to test the hypothesis that a comparison of two methods of minute ventilation, spontaneously generated with mechanically generated, would be a useful predictor of readiness for extubation in preterm infants, weighing <2000 gm, who require mechanical ventilation for >24 hours. STUDY DESIGN: This observational study of 35 infants weighing < or = 2000 gm evaluated the comparison of spontaneously generated minute ventilation with mechanically generated minute ventilation to successfully predict readiness for extubation. After reaching entry criteria, infants were extubated if their spontaneously generated minute ventilation (while receiving endotracheal CPAP) was > or = 50% of the mechanically generated minute ventilation during assist/control ventilation. RESULTS: Of the 35 infants who had a successful trial and were extubated, 30 (86%) remained extubated for at least 24 hours. Of the five infants who failed extubation, four developed apnea and one developed stridor. Thus, a spontaneous minute ventilation of > or = 50% of mechanically generated minute ventilation predicted readiness for extubation in 86% of the patients in this observation. CONCLUSION: A spontaneously generated minute ventilation that is > or = 50% of the mechanically generated minute ventilation is an objective predictor of the readiness for extubation in low birth weight infants who have been weaned to modest ventilatory support. PMID- 9848755 TI - Increased survival in low birth weight neonates given prophylactic surfactant. AB - OBJECTIVE: To compare the effectiveness of a prophylactic surfactant treatment strategy (PRO) to the effectiveness of a rescue (RESC) surfactant treatment strategy in patients at high risk for developing hyaline membrane disease (HMD). STUDY DESIGN: We analyzed data from a retrospective cohort consisting of all patients admitted to the neonatal intensive care units at the centers participating in the recently completed Infasurf-Survanta Comparative Trial. To be in the cohort, a patient had to be admitted during the trial, be <48 hours of age on admission, have a gestational age of <30 weeks, have a birth weight of 501 to 1250 gm, and be free of congenital anomalies. Twelve centers participated in this study. They contributed 1097 patients of whom 381 were treated with a PRO strategy. RESULTS: Survival was significantly higher in the PRO-strategy patients (84% vs 72%, p < 0.05) as was survival without oxygen requirement at a postconceptional age of 36 weeks (60% vs 46%, p < 0.05). In addition, the patients with PRO had a lower prevalence of grade III and IV intraventricular hemorrhage (IVH, 9% vs 14%, p < 0.05). All analyses were controlled for birth weight and type of study center. CONCLUSION: These data support the conclusion that using a PRO treatment strategy results in improved survival in patients at risk for developing HMD. A PRO treatment strategy may also decrease the likelihood of developing a severe IVH. PMID- 9848757 TI - Uterine rupture in women attempting a vaginal birth following prior cesarean birth. AB - OBJECTIVE: To determine (1) if there are any common features to fetal heart rates (FHR) and uterine activity patterns before uterine rupture, (2) if neonatal outcomes as reflected by cord blood gases are associated with the length of fetal bradycardia, and (3) if there is an increase in maternal and/or neonatal length of stay in women who experience uterine rupture during labor as compared with women following repeat cesarean section. METHODS: Maternal and fetal records of 11 women identified by the ICD-9 code as having had a uterine rupture between 1990 and 1995 were retrospectively reviewed. RESULTS: No one common feature in FHR patterns or uterine activity existed before uterine rupture other than bradycardia, although variable and/or late decelerations commonly preceded the bradycardia. Of the neonates, 91% had cord blood pH of <7.0 and 45% had base excess greater than 15 meq/l. Of those fetuses experiencing bradycardia, 55% had placental abruption. Maternal length of stay 5 days or greater occurred in 36% of those with uterine rupture. Seventy-three percent of the neonates required admission to the Neonatal Intensive Care Unit (NICU). Despite acidemia as shown by cord pH, none of the neonates experienced seizures or multiorgan dysfunction. CONCLUSION: There is no one specific FHR or uterine activity pattern that indicates the onset of a uterine rupture, although variable and/or late decelerations occur before the onset of an FHR bradycardia. In the present study, neonatal admissions to the NICU were increased and pH values were below 7.0 in 91% when uterine rupture occurred. A decrease or cessation of uterine tone was not observed. Maternal length of stay was slightly increased following uterine rupture. PMID- 9848758 TI - Endotracheal continuous positive airway pressure after rescue surfactant therapy. AB - OBJECTIVE: To determine if premature infants greater than 31 weeks of gestation with established hyaline membrane disease (HMD) can be treated with endotracheal continuous positive airway pressure (ETCPAP) after rescue surfactant replacement therapy. STUDY DESIGN: Retrospective study of 46 premature infants (>31 weeks of gestation) admitted to Texas Children's Hospital with HMD. Tolerance to ETCPAP after surfactant replacement was evaluated. Prenatal and postnatal characteristics and outcome were compared in the success and failure groups. Multiple logistic regression was used to determine predictive factors associated with failure. RESULTS: Thirty infants (65.2%) were successfully treated with rescue surfactant and ETCPAP. Cesarean section, higher 1-minute Apgar score, and higher FiO2 level at entry were independent predictors of failure to remain on CPAP due to hypoxemia (56.3%), hypercapnia (31.2%), and apnea (12.5%). In the success group duration of intubation (p < 0.001), oxygen administration (p < 0.01), >40% oxygen requirement (p < 0.001), hospital stay (p < 0.05), and respiratory support on day 7 (p < 0.001) were significantly favorable. CONCLUSION: Two thirds of infants greater than 31 weeks of gestation, with HMD needing rescue surfactant treatment, can be successfully managed with ETCPAP. PMID- 9848759 TI - The effects of maternal magnesium sulfate treatment on newborns: a prospective controlled study. AB - OBJECTIVE: To evaluate the effects of maternal magnesium sulfate treatment on newborn outcome. METHODS: Subjects were newborn infants delivered at > or = 34 weeks of gestation whose mothers received a minimum of 12 hours of intravenous MgSO4 therapy before delivery. Control infants were the next born infants of similar gestational age. Outcome recorded at delivery included Apgar scores, whether resuscitation was required, and whether respiratory depression or decreased tone were noted by the physician in attendance. Pneumocardiograms on magnesium-exposed and control infants, obtained within 6 to 18 hours after delivery, were analyzed postdischarge by a single investigator who was blinded to group. The nursery course, feeding patterns, time to first stool, and time to first void were recorded. All patient care decisions, including admission to the neonatal intensive care unit (NICU) or term nursery, were independent of study protocol. RESULTS: A total of 26 magnesium-exposed and 26 control infants were enrolled. The mean total dose of MgSO4 before delivery was 51.2+/-24 gm; the mean duration of therapy was 23.1+/-10 hours. The mean maternal serum magnesium level before delivery was 5.8+/-1.1 mg/dl. The infants' mean cord or initial serum magnesium level was 5.2+/-1.0 mg/dl, which correlated with the maternal magnesium level before delivery (r=0.81, p < 0.001). MgSO4-exposed infants had a higher incidence of hypotonia and lower median Apgar scores than control infants (p < 0.001). However, there was no association between adverse outcomes and maternal serum magnesium concentrations at delivery, duration of treatment, or dose of MgSO4. No difference in dose or length of maternal MgSO4 treatment was noted between infants admitted to term nursery and those admitted to NICU. Pneumocardiogram data were similar between MgSO4-exposed and control infants (all p > or = 0.16). There were no significant differences in number of episodes of feeding intolerance or in time to first stool or void between MgSO4-exposed and control infants (all p > or = 0.31). CONCLUSION: Infants born to mothers treated with MgSO4 were more likely to be hypotonic and have lower Apgar scores at birth. Beyond the immediate postdelivery period, there were no additional complications in this cohort attributable to prenatal MgSO4 exposure. We suggest that pediatricians attend deliveries of magnesium-exposed infants. PMID- 9848760 TI - Clinically significant gastroesophageal reflux in the at-risk premature neonate: relation to cognitive scores, days in the NICU, and total hospital charges. AB - OBJECTIVE: The principal objective of this retrospective, cohort study was to determine if clinically significant gastroesophageal reflux (GER) would impair the long-term cognitive and motor development of preterm infants. An additional objective was to determine the effects of clinically significant GER on the length of hospital stay and total hospital charges in preterm infants. STUDY DESIGN: The design was a retrospective, cohort study of 66 preterm infants, followed in the Neonatal Developmental Follow-Up Clinic of The Children's Hospital in Greenville, SC. Thirty-three premature infants with clinically significant GER met the following study criteria: birth dates, 1988 through 1994; lack of gastrointestinal anatomic defects; and lack of acute neurologic injury (defined as no intraventricular hemorrhage greater than Papile's grade I, no periventricular leukomalacia, no seizures, and no history of birth asphyxia). Clinically significant GER was defined as GER associated with moderate to severe apnea (n=29) or GER associated with moderate to severe feeding intolerance (n=4). The study patients were matched as closely as possible with 33 control premature infants for sex (except when twins were used), ethnicity, social risk, gestational age, birth weight, and Apgar scores at 1 and 5 minutes. Social risk was categorized using Hack's Social Risk Scale. Important covariates included apnea, home monitoring, and nasal continuous positive airway pressure. RESULTS: Neurodevelopmental test scores from 7 months of age through 2 years of age did not show any significant differences between premature infants with clinically significant GER and premature infants with no evidence of clinically significant GER. Total hospital charges were statistically different for the clinically significant GER infants and the nonclinically significant GER infants (median $112,916 versus median $63,928, p=0.01). Total neonatal intensive care unit length of stay measures were statistically different between the two groups (median 53 days versus median 40.5 days, p=0.01). CONCLUSION: Even though clinically significant GER may pose a substantial medical risk in premature infants, the long-term cognitive consequences appear to be negligible. Nevertheless, those premature infants with clinically significant GER do consume significantly more hospital resources than matched controls. Early diagnosis and intervention may possibly lessen the impact of medical costs and reduce length of hospital stay. PMID- 9848761 TI - Thrombolysis of a neonatal brachial artery thrombosis with tissue plasminogen activator. AB - OBJECTIVE: A case report is presented describing the use of tissue plasminogen activator (TPA) to dissolve a thrombosis in the brachial artery of a critically ill premature neonate. A short review of the literature is included. DESIGN: A premature infant with pulmonary hypertension is discovered to have a concomitant right brachial artery thrombosis. A debate ensued concerning the choice of medical management to pursue after surgical extirpation was ruled out. After review of the various alternatives both traditional and novel, a decision was made to utilize TPA. RESULTS: After a continuous infusion of 0.1 mg/kg per hour for 15 hours, blood flow was established distal to the previously occluded right brachial artery. No untoward side-effects occurred during or subsequent to the treatment. CONCLUSION: TPA proved to be an extremely useful medication for the relief of a brachial artery thrombosis in this patient. The theoretical advantages, as well as several anecdotal reports of its successful use, should hopefully stimulate a large multicenter study of its use in the neonatal population. PMID- 9848762 TI - A comparison of two methods of oral endotracheal tube stabilization in neonatal patients. AB - OBJECTIVE: To determine if an endotracheal tube stabilization method using an umbilical clamp reduces accidental extubations when compared with a more traditional method of endotracheal tube stabilization. STUDY DESIGN: From January 1, 1994 through April 30, 1997, the frequency of accidental extubations per 100 ventilator days was tracked for the Neonatal Intensive Care Unit (NICU) Quality Assurance Committee of William Beaumont Hospital (Royal Oak, MI). From January 1, 1994 through December 31, 1995, all endotracheal tubes were stabilized using a traditional taping method. From March 1, 1996 through April 30, 1997, the endotracheal tubes were stabilized using a modified umbilical clamp and tape. The incidence of extubation using each stabilization method was compared by chi2 analysis. RESULTS: The umbilical clamp resulted in a significantly lower (p=0.001) rate of accidental extubations. CONCLUSION: The umbilical clamp taping method appears to have lowered the incidence of accidental extubation in our NICU when compared with a standard method of oral endotracheal tube stabilization. PMID- 9848763 TI - Intraperitoneal administration of cytomegalovirus hyperimmunoglobulin to the cytomegalovirus-infected fetus. AB - Twenty-five percent of cytomegalovirus (CMV)-infected fetuses had sequelae and 8% of those in the recurrent-infected group had sequelae. There is no report yet on the fetal therapy for CMV infections. A Japanese pregnant woman with intrauterine fetal CMV infection diagnosed at 26 weeks of pregnancy is presented. CMV culture of amniotic fluid was positive. A CMV DNA assay using the polymerase chain reaction method of the cord blood and the amniotic fluid was positive during the pregnancy; however, testing for fetal serum CMV-specific IgM was negative. The CMV IgG titer of fetal serum at 27 weeks of pregnancy was a third of that of the maternal serum. CMV hyperimmunoglobulin was injected into the fetal abdominal cavity at 28 and 29 weeks of pregnancy. A second administration of CMV hyperimmunoglobulin increased the titer of CMV IgG in the fetal circulation. At birth, the urine culture was positive for CMV. However, CMV DNA of the ascites became negative. A brain CT scan performed 2 weeks after birth revealed some small calcifications beside the right ventricle. CMV hyperimmunoglobulin injection to the fetal abdominal cavity has been shown to increase the IgG in the fetal serum. This is the first report of fetal therapy of congenital CMV infection. PMID- 9848764 TI - The inadequate male syndrome. PMID- 9848765 TI - The development of newborn care in the UK since 1930: AAP Thomas E. Cone, Jr. Lecture on Perinatal History. PMID- 9848767 TI - Early neonatal urinary tract infection: a case report and review. AB - We report the earliest case of urinary tract infection (UTI) so far described in term uncomplicated newborn infants. The unusual features of this case included early massive hematuria as the only presenting symptom and a host with no risk factors for either sepsis or localization of infection in the genitourinary tract. Reports of urinary analysis, urine cultures, renal ultrasound, and renal scan established the diagnosis of UTI. Massive hematuria in this case was probably precipitated by mild renal venous thrombosis. Ampicillin-resistant Escherichia coli was cultured from the urine, which responded promptly to appropriate treatment. A urine culture was done because of the predominantly urinary signs and symptoms. Although urine culture analysis is not routinely performed for the evaluation of possible sepsis before 72 hours of age, this investigation may be important in critically ill neonates who present with predominantly genitourinary signs and symptoms. PMID- 9848766 TI - Favism by proxy in nursing glucose-6-phosphate dehydrogenase-deficient neonates. AB - Two nursing neonates deficient in glucose-6-phosphate dehydrogenase developed severe hyperbilirubinemia despite phototherapy. Mothers of both the infants had recently eaten fava beans. The hemolytic triggers found in fava beans may have been absorbed by the mothers and excreted in their breast milk. Carboxyhemoglobin determination performed on one of the infants reflected ongoing hemolysis. PMID- 9848768 TI - Ventilatory casebook. Late presentation of congenital diaphragmatic hernia in association with group B streptococcal sepsis and meningitis. PMID- 9848769 TI - Radiology casebook. Congenital toxoplasmosis: a case report. PMID- 9848770 TI - Special imaging casebook. Tetralogy of Fallot with absent pulmonary valve syndrome and partial DiGeorge syndrome. PMID- 9848771 TI - Resuscitation of a micropremie: the case of MacDonald versus Milleville. PMID- 9848772 TI - Decreased aquaporin-2 expression and apical plasma membrane delivery in kidney collecting ducts of polyuric hypercalcemic rats. AB - Hypercalcemia is frequently associated with a urinary concentrating defect and overt polyuria. The molecular mechanisms underlying this defect are poorly understood. Dysregulation of aquaporin-2 (AQP2), the predominant vasopressin regulated water channel, is known to be associated with a range of congenital and acquired water balance disorders including nephrogenic diabetes insipidus and states of water retention. This study examines the effect of hypercalcemia on the expression of AQP2 in rat kidney. Rats were treated orally for 7 d with dihydrotachysterol, which produced significant hypercalcemia with a 15 +/- 2% increase in plasma calcium concentration. Immunoblotting and densitometry of membrane fractions revealed a significant decrease in AQP2 expression in kidney inner medulla of hypercalcemic rats to 45.7 +/- 6.8% (n = 11) of control levels (100 +/- 12%, n = 9). A similar reduction in AQP2 expression was seen in cortex (36.9 +/- 4.2% of control levels, n = 6). Urine production increased in parallel, from 11.3 +/- 1.4 to a maximum of 25.3 +/- 1.9 ml/d (P < 0.01), whereas urine osmolality decreased from 2007 +/- 186 mosmol/kg x H2O to 925 +/- 103 mosmol/kg x H2O (P < 0.01). Immunocytochemistry confirmed a decrease in total AQP2 labeling of collecting duct principal cells from kidneys of hypercalcemic rats, and reduced apical labeling. Immunoelectron microscopy demonstrated a significant reduction in AQP2 labeling of the apical plasma membrane, consistent with the development of polyuria. In summary, the results strongly suggest that AQP2 downregulation and reduced apical plasma membrane delivery of AQP2 play important roles in the development of polyuria in association with hypercalcemia. PMID- 9848773 TI - Chloride transport by the rabbit cortical collecting duct: dependence on H,K ATPase. AB - The rabbit cortical collecting duct (CCD) exhibits the capacity for active chloride absorption when basolateral Na-K-ATPase is inhibited by ouabain. The present studies examine the contribution of H,K-ATPase to this ouabain insensitive Cl absorption and to related ion fluxes. Rabbits were fed a KCl-rich diet with no measurable Na for 4 to 13 d before isolation of the CCD for microperfusion. Application of peritubular ouabain (0.1 mM) significantly increased (P < 0.001) net luminal absorptive chloride flux (J(N)Cl) without an effect on lumen-to-bath isotopic 36Cl flux (J(lb)Cl). The H,K-ATPase inhibitor Sch 28080 (1 to 10 microM) abolished ouabain-insensitive J(N)Cl, but transepithelial voltage (V(T)) was not significantly affected. The contribution of H,K-ATPase activity on active Cl flux (J(A)Cl) and passive Cl flux (J(P)Cl) was also assessed. Ouabain significantly increased J(A)Cl and Sch 28080 inhibited J(A)Cl, but J(P)Cl was not affected by Sch 28080. To assess the contribution of changes in net bicarbonate flux (JtCO2) to changes in J(N)Cl, JtCO2 was measured under identical conditions as for J(N)Cl. Ouabain significantly increased JtCO2, and this ouabain-insensitive bicarbonate flux was inhibited by Sch 28080 without significantly affecting V(T). To assess the possibility that the CCD may possess mechanisms for neutral salt absorption, lumen-to-bath 86Rb efflux (K(Rb)), and 22Na efflux (K(Na)) were also measured. Ouabain significantly increased K(Rb), and Sch 28080 inhibited this ouabain-insensitive K(Rb). Furthermore, Sch 28080 and A80915a (a structurally distinct H,K-ATPase inhibitor) significantly inhibited K(Na) in the presence of 1 mM luminal amiloride. These observations suggest that, in addition to potassium, sodium can be transported via the H,K ATPase. Although the CCD contains more than one cell population, the data could be fitted very well to the function of the B-type intercalated cell. A cell model is proposed for the hypothesis that ouabain-insensitive chloride absorption is mediated by the parallel operation of an apical H,K-ATPase with an apical Cl-HCO3 exchanger and that the H,K-ATPase can function, under certain conditions, as a mechanism of Na absorption. PMID- 9848774 TI - Amiloride-insensitive Na+-H+ exchange: a candidate mediator of erythrocyte Na+ Li+ countertransport. AB - Erythrocyte Na+-Li+ countertransport shows an increased activity in essential hypertension and diabetic nephropathy, but its nature remains unknown. This amiloride-insensitive membrane transport may not be a mode of operation of the amiloride-sensitive NHE1, the only Na+-H+ exchange isoform found in human erythrocytes. Whether an independent, although unknown, amiloride-insensitive isoform mediates Na+-Li+ countertransport is unclear. Na+-H+ exchange activity was measured in acid-loaded erythrocytes. Dimethylamiloride, a specific inhibitor of Na+-H+ exchange and phloretin, a known inhibitor of Na+-Li+ countertransport, gave a reduction in H+-driven Na+ influx (by 31 and 37%, respectively). This effect was additive, and a 66% reduction in H+-driven Na+ influx was found in the presence of both inhibitors. Internal acidification, a stimulus for Na+-H+ exchange, enhanced Na+-Li+ countertransport activity (from 287 +/- 55 to 1213 +/- 165 micromol x Lcell(-1) h(-1), mean +/- SEM, P = 0.003). This transport remained sensitive to phloretin under both conditions. Conversely, external acidification decreased Na+-Li+ countertransport activity (as expected for a Na+-H+ exchanger). Competition between internal H+ and Li+ or Na+ for a common binding site was present. Finally, similar kinetic parameters for external Na+ characterized Na+ Li+ countertransport and the phloretin-sensitive component of H+-driven Na+ influx. These findings suggest that both Na+-Li+ countertransport and the amiloride-insensitive, phloretin-sensitive component of H+-driven Na+ influx can be mediated by a previously unrecognized novel amiloride-insensitive Na+-H+ exchange isoform in human erythrocytes. PMID- 9848775 TI - Pressure-natriuresis and -diuresis in transgenic rats harboring both human renin and human angiotensinogen genes. AB - The hypertensive double transgenic rat harboring both the human renin and human angiotensinogen genes (dTGR) offers a unique opportunity to study the human renin angiotensin system in an experimental animal model. Since nothing is known about the control of sodium and water excretion in these rats, this study was performed to compare pressure-natriuresis relationships in hypertensive dTGR and normotensive control rats harboring only the human renin gene (hREN), in order to determine how the pressure-natriuresis relationship is reset in hypertensive dTGR. To differentiate between extrinsic and intrinsic renal mechanisms, experiments were performed with and without renal denervation, and with and without infusions of vasopressin, norepinephrine, 17-OH-corticosterone, and aldosterone. Human and rat angiotensinogen and renin mRNA expression were also determined. In hREN without controlled renal function, urine flow and sodium excretion increased from 13 to 169 microl/min per g kidney wet weight (kwt) and from 1 to 30 micromol/min per g kwt, respectively, as renal perfusion pressure was increased from 67 to 135 mmHg. Renal blood flow (RBF) and GFR ranged between 3 to 7 and 0.9 to 1.5 ml/min per g kwt. In dTGR, pressure-natriuresis-diuresis relationships were shifted approximately 40 mmHg rightward. RBF was lower in dTGR than in hREN; GFR was not different. In dTGR with neurohormonal factors controlled, RBF was decreased and pressure-natriuresis-diuresis curves were not different compared to dTGR curves without these interventions. By light microscopy, the kidneys of these 6-wk-old dTGR and hREN rats were normal and indistinguishable. Both human and rat renin and angiotensinogen mRNA were expressed in the kidneys of dTGR. The two renin mRNA were decreased in dTGR, indicating a physiologic downregulation of renin gene expression by high BP. It is concluded that the renal pressure-natriuresis mechanism is reset toward higher pressure levels in dTGR and participates in the maintenance of hypertension. The reduced excretory function in dTGR depends on hREN and human angiotensinogen gene expression and is intrinsic to the kidney as opposed to extrarenal regulators. PMID- 9848776 TI - Effects of the angiotensin II antagonist valsartan on blood pressure, proteinuria, and renal hemodynamics in patients with chronic renal failure and hypertension. AB - Angiotensin II receptor antagonists have become clinically available for the treatment of arterial hypertension. Presently, there is little information about their effects on BP, proteinuria, and renal function in patients with moderate or advanced renal failure. This study examines the effects of the angiotensin II antagonist Valsartan (80 mg/d) on proteinuria and glomerular permselectivity in patients with chronic renal failure during a 6-mo treatment, using a double blind, randomized, placebo-controlled study (treatment group [V-group]: n = 5, age 57 +/- 7 yr, serum creatinine 365 +/- 122 micromol/L; placebo group [P group]: n = 4, age 62 +/- 11 yr, serum creatinine 346 +/- 61 micromol/L). Study parameters included BP, 24-h proteinuria, GFR, and effective renal plasma flow (ERPF) as determined by inulin and para-aminohippurate clearance. Changes in glomerular permselectivity were assessed by measuring the fractional clearances of neutral dextrans by HPLC gel-permeation chromatography. Valsartan lowered the mean arterial pressure on average by 13 +/- 7 mmHg during the 6-mo treatment (P < 0.05). GFR and ERPF remained almost unchanged. However, after 6 mo of Valsartan treatment, proteinuria was reduced by 396 +/- 323 mg/24 h (26 +/- 18%) and albuminuria by 531 +/- 499 mg/24 h (41 +/- 21%) with regard to baseline values (P < 0.05). In the P-group, both proteinuria and albuminuria increased slightly with time (by 30 +/- 43% and 30 +/- 54%, respectively, NS). The fractional clearances of high molecular weight dextrans >66 A were significantly reduced after 6 mo of Valsartan treatment (P < 0.05), indicating a reduction of the glomerular shunt volume by 54 +/- 20% (P < 0.05) according to the model of Deen et al. (Am J Physiol 249: 347-389, 1985). The mean pore size radius of the glomerular membrane remained unchanged. This effect was independent of glomerular hemodynamic changes. Valsartan persistently lowered proteinuria in patients with chronic renal failure. Although GFR and ERPF remained nearly stable, this effect could be attributed to an improvement in glomerular permselectivity. PMID- 9848777 TI - Lovastatin inhibits mesangial cell proliferation via p27Kip1. AB - Mesangial cell proliferation is a key feature of glomerulonephritis. The hydroxymethylglutaryl-coenzyme A reductase inhibitor lovastatin is known to inhibit cell cycle progression. To determine the inhibitory mechanisms of mesangial cell proliferation by lovastatin, the cyclin-dependent kinase (CDK) activity, and expression of CDK inhibitor (p27Kip1, p21Cip1, and p16INK4) mRNA and protein were measured. Lovastatin inhibited phosphorylation of retinoblastoma protein and mesangial cell proliferation dose dependently. Lovastatin increased the p27Kip1 protein level but produced no changes in the abundance of the p27Kip1 mRNA level both in the presence and absence of mitogens. Treatment with lovastatin revealed the increment of both CDK2- and CDK4-bound-p27Kip1. The experiment using antisense oligonucleotide against p27Kip1 showed significant amelioration of lovastatin-induced cell cycle arrest. Lovastatin reduced both platelet-derived growth factor-stimulated CDK2 and CDK4 kinase activities. In conclusion, lovastatin inhibited mesangial proliferation via translational upregulation or impairment of p27Kip1 protein degradation. Lovastatin serves as a potential therapeutic approach to mesangial proliferative disease. PMID- 9848778 TI - Hepatitis C viral infection is associated with fibrillary glomerulonephritis and immunotactoid glomerulopathy. AB - The most common form of glomerular disease seen in association with hepatitis C virus (HCV) infection is membranoproliferative glomerulonephritis, with or without associated cryoglobulinemia. This study examines four cases of fibrillary glomerulonephritis and two cases of immunotactoid glomerulopathy in association with HCV infection. Findings at presentation included proteinuria, renal insufficiency, and hematuria. Renal biopsy revealed a membranoproliferative pattern of glomerular disease in five cases, and a membranous glomerulopathy with mesangial proliferative features in one. On immunofluorescence, all cases stained with IgG and C3. Electron microscopy revealed fibrils of the expected diameter, 16 to 28 nm in fibrillary glomerulonephritis and 33 to 45 nm in immunotactoid glomerulopathy. In only one case were cryoglobulins detected (at low titer and on only one of three assays). Antiviral therapy was not given in any of the six cases. Outcomes were mixed, with progression to renal failure occurring in two patients and persistent proteinuria with stable or improved renal function in three. Follow-up is not available on the sixth case. Both fibrillary glomerulonephritis and immunotactoid glomerulopathy have features that overlap with cryoglobulinemic glomerulonephritis. The relatedness of these three entities in a subset of patients with HCV infection suggests a common pathogenic mechanism of glomerular deposition of organized deposits. PMID- 9848779 TI - Expression of types I, II, and III TGF-beta receptors in human glomerulonephritis. AB - Protein and mRNA expression of transforming growth factor-beta (TGF-beta) receptor type I (TbetaRI), type II (TbetaRII), and type III (TbetaRIII) were studied in serial sections of kidney samples obtained from patients with glomerulonephritis. In minimal change disease, weak expression of TbetaRI and TbetaRII was observed mainly in glomerular endothelial cells, peritubular capillaries, and interstitial arteriolar endothelial cells, whereas TbetaRIII expression was found mainly in the interstitium. Expression of all three TGF-beta receptors (TbetaR) was increased remarkably in glomerular and Bowman's capsular cells comprising the tuft adhesions to Bowman's capsules in glomerulonephritis with increased matrix accumulation, including IgA nephropathy, lupus nephritis, focal and segmental glomerulosclerosis, myeloperoxidase-antineutrophil cytoplasmic antibody-associated crescentic glomerulonephritis, and membranoproliferative glomerulonephritis. Increased expression of the three TbetaR was also seen in glomerular epithelial cells in the vicinity of glomerulosclerotic lesions, in crescent cells, and in some tubules and infiltrative mononuclear cells found in the periglomerular and tubulointerstitial lesions with increased matrix deposition. In contrast, no remarkable TbetaRII expression was noted in mesangial proliferative lesions in IgA nephropathy, lupus nephritis, and membranoproliferative glomerulonephritis. These data suggest that distinctive modulation of TbetaR expression may be involved in the development of adhesive, sclerotic, and proliferative renal lesions in human glomerulonephritis. PMID- 9848780 TI - Enhanced expression of membrane type-1 matrix metalloproteinase in mesangial proliferative glomerulonephritis. AB - Matrix metalloproteinase-2 (MMP-2, gelatinase A) is involved in the inflammatory and sclerotic events of glomerular diseases. Newly identified membrane-type matrix metalloproteinases (MT-MMP) have been shown to activate specifically proMMP-2. To date, several types of MT-MMP have been cloned; however, their expressions in glomerular diseases have not been evaluated. To investigate the role of MT-MMP in glomerular diseases, the glomerular gene expression and enzymatic activity of MT-MMP were examined during the time course of nephritis induced in rats by anti-Thy1.1 antibody injection. Both MT1-MMP and MMP-2 mRNA expression increased prominently 5 and 10 d after anti-Thy1.1 antibody injection and decreased thereafter, as assayed by semiquantitative reverse transcription PCR. In contrast, there were no remarkable changes in the gene expression of MT2 MMP between normal and diseased tissue, and that of MT3-MMP was not detected in isolated glomeruli by reverse transcription-PCR analysis. The activation of proMMP-2 as analyzed by gelatin zymography correlated with the glomerular MT1-MMP gene expression, suggesting that proMMP-2 was activated by MT1-MMP. Protein and mRNA expression of fibronectin, one of the major mesangial matrix proteins and substrate of MMP-2, were also synchronized with MT1-MMP and MMP-2 expression. In situ hybridization revealed intense MT1-MMP mRNA expression in the proliferating mesangial cells. Interestingly, MT1-MMP gene expression exhibited a similar distribution as alpha-smooth muscle actin expression, which was closely associated with mesangial phenotypic change. These results suggest that among the newly identified MT-MMP, MT1-MMP may play the central role in activation of proMMP-2. Furthermore, the enhancement of MT1-MMP and MMP-2 expression associated with mesangial phenotypic change may contribute to the development of anti-Thy1.1 antibody-induced glomerulonephritis and remodeling of extracellular matrices. PMID- 9848781 TI - Mycophenolate mofetil prevents the induction of active Heymann nephritis: association with Th2 cytokine inhibition. AB - The effect of mycophenolate mofetil (MMF) was examined in active Heymann nephritis (HN), an animal model of human membranous nephropathy. HN was induced in Lewis rats with Fx1A/complete Freund's adjuvant (CFA), and controls only received CFA. The induction of HN was prevented by MMF (30 mg/kg per d) from 0 to 4 wk after immunization. Proteinuria was not different in CFA controls up to 16 wk, and was significantly less than in untreated HN from 6 wk onward. Serum anti Fx1A antibody (Ab) levels and glomerular Ig deposition were suppressed during therapy. The interstitial infiltrate of alphabetaTCR+, CD4+ and CD8+ T cells, natural killer cells, and macrophages (mphi) observed in untreated HN at 8 wk was absent from rats treated from 0 to 4 wk with MMF. Semiquantitative reverse transcription-PCR for renal mononuclear cell cytokine mRNA at 8 wk demonstrated that MMF from 0 to 4 wk prevented the increased expression of Th1 (interferon gamma, lymphotoxin), Th2 (interleukin-4), and mphi (tumor necrosis factor-alpha) cytokines identified in untreated HN. In lymph node draining sites of immunization, MMF limited both enlargement and the increased proportion of CD3+, CD4+, and CD8+ T cells observed in untreated HN and CFA controls. MMF suppressed Th2 (interleukin-4) but not Th1 (interferon-gamma, lymphotoxin) cytokine mRNA expression in lymph nodes. MMF from 4 to 8, 6 to 12, or 10 to 14 wk did not prevent proteinuria, serum anti-Fx1A Ab, or glomerular IgG deposition when compared with untreated HN. This study showed that MMF from 0 to 4 wk prevented the induction of HN and was associated with preferential suppression of Th2 cytokines. This therapy may prove useful in human idiopathic membranous nephropathy. PMID- 9848782 TI - Hyaluronan induces monocyte chemoattractant protein-1 expression in renal tubular epithelial cells. AB - Hyaluronan (HA) is a nonsulfated glycosaminoglycan that accumulates in the renal interstitium in immune-mediated kidney diseases. The functional significance of such HA deposition in the kidney has not been elucidated. Several studies have suggested that HA may exhibit proinflammatory effects. Since chemokines such as monocyte chemoattractant protein-1 (MCP-1) play an important role in the recruitment of leukocytes in renal injury, this study tested whether HA and its fragments could promote MCP-1 production by renal parenchymal cells. Mouse cortical tubular cells were stimulated with fragmented HA or with high molecular weight HA (Healon) in vitro and were examined for MCP-1 expression. Fragmented HA, but not Healon, increased MCP-1 mRNA within 30 min with a peak after 2 h. In addition, a 10-fold increase of MCP-1 protein in the supernatant was found after a 6-h stimulation with fragmented HA. The enhanced MCP-1 mRNA and protein expression in response to HA was dose-dependent between 1 and 100 microg/ml. Upregulation of MCP-1 protein production could be blocked by preincubation with actinomycin D or cycloheximide, suggesting that MCP-1 mRNA and protein expression in response to HA are based on de novo synthesis. The HA-stimulated MCP-1 production was also inhibited with anti-CD44 antibodies, suggesting that MCP-1 is upregulated at least in part by signaling through CD44. In summary, fragmented HA markedly stimulates renal tubular MCP-1 production by mechanisms that involve binding to the HA receptor CD44. It is hypothesized that the accumulation of HA in immune renal injury could participate in the recruitment and activation of inflammatory cells in vivo through production of MCP-1. PMID- 9848783 TI - High mutation detection rate in the COL4A5 collagen gene in suspected Alport syndrome using PCR and direct DNA sequencing. AB - Approximately 85% of patients with Alport syndrome (hereditary nephritis) have been estimated to have mutations in the X chromosomal COL4A5 collagen gene; the remaining cases are autosomal with mutations in the COL4A3 or COL4A4 genes located on chromosome 2. In the present work, the promoter sequence and previously unknown intron sequences flanking exons 2 and 37 of COL4A5 were determined. Furthermore, intron sequences flanking the other 49 exons were expanded from 35 to 190 to facilitate mutation analysis of the gene. Using this information, all 51 exons and the promoter region were PCR-amplified and sequenced from DNA of 50 randomly chosen patients with suspected Alport syndrome. Mutations were found in 41 patients, giving a mutation detection rate of 82%. Retrospective analysis of clinical data revealed that two of the cases might be autosomal. Although it could not be determined whether the remaining seven cases (14%) were autosomal or X chromosome-linked, it is likely that some of them were autosomal. It is concluded that PCR amplification and direct DNA sequencing of the promoter and exons is currently the best procedure to detect mutations in COL4A5 in Alport syndrome. PMID- 9848784 TI - TGF-beta1 gene mutations in insulin-dependent diabetes mellitus and diabetic nephropathy. AB - PCR assays were established for easy and fast analysis of two transforming growth factor-beta1 (TGF-beta1) gene mutations, a C to T transition at position 76 in exon 5 resulting in a change from threonine to isoleucine in position 263 (Thr263Ile) of the propeptide and a deletion of a C in the intron sequence eight bases prior to exon 5 (713-8delC). These mutations were evaluated in insulin dependent diabetes mellitus (IDDM) patients (n = 137) and control subjects (n = 105) and in IDDM patients with (n = 170) and without (n = 99) nephropathy. After evaluating intra- and interindividual variation in TGF-beta1 expression levels, the TGF-beta1 mRNA level in phorbol 12-myristate-13-acetate-stimulated (1 ng/ml) lymphocytes from individuals with different TGF-beta1 genotypes was also studied. No association of the two TGF-beta1 sequence variations with IDDM in general was found. However, a weak but significant association of the Thr263Ile mutation with diabetic nephropathy was found (P = 0.03). No correlation between TGF-beta1 transcription level and genotype of any of the two studied polymorphisms was found. However, significant interindividual differences in TGF-beta1 mRNA levels were observed between the tested individuals (P < 0.0001) compatible with a genetic control mechanism of TGF-beta1 synthesis at the mRNA level. PMID- 9848785 TI - The antiproteinuric effect of angiotensin antagonism in human IgA nephropathy is potentiated by indomethacin. AB - Evidence is available from animal and human studies that protein traffic through the glomerular capillary has a pathogenetic role in subsequent renal damage and that angiotensin-converting enzyme (ACE) inhibitors appear superior to other drugs in lowering proteinuria and the rate of renal function decline. This study compares the effect of ACE inhibition or angiotensin II (AngII) receptor blockade on urinary protein excretion and renal hemodynamics in 20 patients with IgA glomerulonephritis randomized to receive enalapril (20 mg/d) or irbesartan (100 mg/d) for 28 d in a double-blind study with two parallel groups. This study also evaluated whether addition of indomethacin (75 mg twice a day) to each of the two treatments resulted in a more potent antiproteinuric effect. Enalapril alone reduced total protein excretion (61% change from baseline) and fractional clearance of albumin without changes in GFR and minor elevation in renal plasma flow. Also, patients randomized to receive the AngII receptor antagonist irbesartan for 28 d had lower proteinuria (55% change from baseline) and fractional clearance of albumin at the end of the treatment period with similar renal hemodynamic changes. When indomethacin was added to enalapril treatment, a further significant reduction in urinary proteins and fractional albumin clearance was observed. In patients given irbesartan, the addition of indomethacin further reduced proteinuria and fractional clearance of albumin. The combined therapy with enalapril or irbesartan and indomethacin did not significantly affect GFR and renal plasma flow compared with baseline. These findings indicate that in patients with IgA glomerulonephritis the antiproteinuric effect of blocking AngII activity by either ACE inhibitors or AngII receptor antagonists is potentiated by indomethacin, an effect that occurred without impairment of renal function. PMID- 9848786 TI - Human complement factor H deficiency associated with hemolytic uremic syndrome. AB - This study reports on six cases of deficiency in the human complement regulatory protein Factor H (FH) in the context of an acute renal disease. Five of the cases were observed in children presenting with idiopathic hemolytic uremic syndrome (HUS). Two of the children exhibited a homozygous deficiency characterized by the absence of the 150-kD form of Factor H and the presence, upon immunoblotting, of the 42-kD Factor H-like protein 1 (FHL-1) and other FH-related protein (FHR) bands. Southern blot and PCR analysis of DNA of one patient with homozygous deficiency ruled out the presence of a large deletion of the FH gene as the underlying defect for the deficiency. The other four children presented with heterozygous deficiency and exhibited a normal immunoblotting pattern of proteins of the FH family. Factor H deficiency is the only complement deficiency associated with HUS. These observations suggest a role for FH and/or FH receptors in the pathogenesis of idiopathic HUS. PMID- 9848787 TI - HIV-associated nephropathy: outcome and prognosis factors. Groupe d' Etudes Nephrologiques d'Ile de France. AB - Records of 102 patients with biopsy-proven HIV-associated nephropathy (HIVAN) admitted to 18 hospitals in the Paris area from 1984 through 1996 were retrospectively reviewed. Demographics and clinical and laboratory features of the cohort were determined, and prognostic factors of renal and patient survival were analyzed. Renal and patient survival curves were estimated with the actuarial method. Prognostic factors were assessed by uni- and multidimensional analyses based on Cox regression models. Values were expressed as median with interquartile. The total population (median age 34) included 97% blacks and 71.5% males. Median patient follow-up was 165 d (range, 43 to 493). At the time of renal biopsy, median values of serum creatinine, proteinuria, and CD4+ cell count were 496 micromol/L, 6.5 g/24 h, and 48.5 cells/mm3, respectively. Fifteen patients were given steroids after the onset of HIVAN. Overall patient survival at 0.5, 1, and 3 yr was 73 +/- 5, 55 +/- 6, and 38 +/- 7%, respectively. The proportion of patients free of dialysis at 0.5, 1, and 3 yr was 73 +/- 5, 60 +/- 7, and 18 +/- 10%, respectively. Predictors of poor patient prognosis were a low CD4+ cell count (relative risk [RR; per 50 cells/mm3 decrease] 1.35; confidence interval [CI], 1.13 to 1.6) and antiretroviral therapy before the onset of HIVAN (RR 1.9; CI, 1.05 to 3.6). Main independent factors associated with better renal outcome were: steroid therapy (RR 0.29; CI, 0.1 to 0.9); low proteinuria level (RR [per 50% decrease] 0.7; CI, 0.5 to 0.98); low serum creatinine (RR [per 1.1 mg/dl decrease] 0.78; CI, 0.7 to 0.87); and hemoglobin level (RR [per g/dl increase] 0.76; CI, 0.58 to 1.00). HIVAN is not a rare nephropathy in Paris and its suburbs. Renal prognosis and patient survival are better than what was reported previously. Steroids may delay the downward course of HIVAN. It is not certain that in the new era of HIV therapy, the possible renal benefits of corticosteroids outweigh their potential risks. The only reliable predictor of patient survival is the intensity of immunodeficiency. PMID- 9848788 TI - The nephropathy of non-insulin-dependent diabetes: predictors of outcome relative to diverse patterns of renal injury. AB - Nephropathy of non-insulin-dependent diabetes mellitus (NIDDM) is the most common cause of end-stage renal failure (ESRF) in Western countries. This study investigates the clinical and histologic putative predictors of disease progression, with the final goal to identify patients at risk who may benefit from early diagnosis and intervention. It examines by repeated measurements of BP, blood glucose, serum creatinine, and urinary protein excretion rate 65 consecutive NIDDM patients with clinical, persistent proteinuria and biopsy documented typical diabetic glomerulopathy (class I; n = 30), predominant nephroangiosclerosis (class II; n = 23), or nondiabetic type glomerulopathy (class III; n = 12), whose severity of renal tissue involvement was precisely quantified by a global histologic score. Baseline parameters and progression to renal end points, i.e., doubling of baseline serum creatinine, dialysis, or transplantation, were univariately and multivariately correlated by proportional hazards regression models. The median kidney survival time in the overall study population was 3.07 yr. Thirty-seven percent of patients reached an end point during a median (range) follow-up of 1.8 yr (0.4 to 5.7 yr). By univariate and multivariate analysis, kidney survival significantly correlated with baseline urinary protein excretion rate (P = 0.04 and P = 0.04, respectively) and renal tissue injury score (P = 0.0001 and P = 0.02, respectively), but not with the histologic classes. Patients with a urinary protein excretion rate < or = 2 g/24 h, or > 2 g/24 h with a histologic score < 7, never reached an end point. All patients with urinary protein excretion > 2 g/24 h and a histologic score > 13 progressed to ESRF over a median of 1.6 yr. No differences in other baseline parameters or in BP and diabetes control during follow-up accounted for these different outcomes. In NIDDM as well as in nondiabetic chronic renal disease, quantification of urinary protein excretion rate--independent of the pattern of underlying glomerular involvement--reliably discriminates progressors from nonprogressors and, combined with precise quantification of renal tissue injury, reliably predicts risk of ESRF. This information may be used to set guidelines for early diagnosis and appropriate intervention to reduce the number of diabetic patients who will need renal replacement therapy in years to come. PMID- 9848789 TI - Pharmacokinetics of alphaIFN-2b in chronic hepatitis C virus patients undergoing chronic hemodialysis or with normal renal function: clinical implications. AB - In this prospective controlled study, the pharmacokinetic profiles of alpha interferon 2b (alphaIFN-2b) were determined by the enzyme-linked immunosorbent assay method in hepatitis C virus-positive (HCV+) dialysis and nonuremic patients, after a single subcutaneous injection of 3 million units. Ten HCV+/RNA+ patients (group A) with a normal renal function (mean serum creatinine: 1.03 +/- 0.26 [SD] mg/dl) and 10 HCV+/RNA+ patients undergoing chronic hemodialysis (group B) were included. The pharmacokinetic profiles of alphaIFN were determined after the very first subcutaneous injection of the drug. Plasma alphaIFN concentrations were determined before the injection and then 1, 2, 3, 4, 6, 8, 12, 16, 20, 24, 28, 32, and 36 h after the injection. They were assessed by means of an enzyme linked immunosorbent assay test. Patients from both groups had a similar body surface area. It was found that in group B: (1) the mean maximum (SD) serum alphaIFN concentration (Cmax) was significantly higher (52 +/- 12 pg/ml) than in group A (39 +/- 12 pg/ml; P = 0.03); (2) the time at which Cmax occurred (Tmax) was significantly higher (10 +/- 3 h) than in group A (7.5 +/- 2 h; P = 0.05); (3) the observed area under the plasma alphaIFN concentration-time curve was about twice as much, i.e., 936 +/- 212 pg x h/ml, as that for group A (485 +/- 184 pg x h/ml; P < 0.0001); and (4) the alphaIFN half-life was significantly longer (9.6 +/- 2.9 h) than in group A (5.3 +/- 1.3 h). As early as 24 h after the alphaIFN injection was given, the drug was no longer detectable in nonuremic patients' sera, whereas it could be detected up to the next injection in all of the dialysis patients' sera. When trough levels of alphaIFN were measured just before the 10th injection, they were always below the threshold level in the 10 patients from group A, i.e., 4.1 pg/ml, whereas in group B they were measurable for four of nine patients (P = 0.05) and ranged between 5.8 and 36.1 pg/ml. Severe neurologic side effects were observed only in group B, i.e., in three patients. Hemoglobin levels did significantly decrease but only in group B patients, and this was significantly correlated with the Cmax (r = 0.67; P = 0.03). This is the first controlled study to demonstrate that the clearance of alphaIFN is about twice as low in dialysis patients as in nonuremic patients. These results might be of relevance when deciding the optimal alphaIFN therapy scheme for HCV+ patients, either with normal renal function or undergoing chronic hemodialysis. PMID- 9848791 TI - Large deletion of the 5' end of the ROMK1 gene causes antenatal Bartter syndrome. AB - Mutations in exon 5 of the ROMK1 gene (KCNJ1) have recently been observed in antenatal Bartter syndrome patients. This study describes a homozygous deletion removing KCNJ1 exons 1 and 2 observed in a consanguineous family with antenatal Bartter syndrome. Absence of the untranslated exon 1 led to the deletion of transcription elements located in this exon that may cause the disease. Deletion of exon 1 transcription elements should lead to the absence of hROMK2-K5 transcripts, whereas hROMK1 transcripts should normally be transcripted. Consequently, probably only hROMK2-K5 transcripts are expressed in the medullary thick ascending limb of Henle. PMID- 9848792 TI - The art of nature: urine under the microscope. PMID- 9848790 TI - Accumulation of carbonyls accelerates the formation of pentosidine, an advanced glycation end product: carbonyl stress in uremia. AB - Advanced glycation end product (AGE) formation is related to hyperglycemia in diabetes but not in uremia, because plasma AGE levels do not differ between diabetic and nondiabetic hemodialysis patients. The mechanism of this phenomenon remains elusive. Previously, it was suggested that elevation of AGE levels in uremia might result from the accumulation of unknown AGE precursors. The present study evaluates the in vitro generation of pentosidine, a well identified AGE structure. Plasma samples from healthy subjects and nondiabetic hemodialysis patients were incubated under air for several weeks. Pentosidine levels were determined at intervals by HPLC assay. Pentosidine rose to a much larger extent in uremic than in control plasma. Pentosidine yield, i.e., the change in pentosidine level between 0 and 4 wk divided by 28 d, averaged 0.172 nmol/ml per d in uremic versus 0.072 nmol/ml per d in control plasma (P < 0.01). The difference in pentosidine yield between uremic and control plasma was maintained in samples ultrafiltrated through a filter with a 5000-Da cutoff value and fortified with human serum albumin (0.099 versus 0.064 nmol/ml per d; P < 0.05). Pentosidine yield was higher in pre- than in postdialysis plasma samples (0.223 versus 0.153 nmol/ml per d; P < 0.05). These results suggest that a large fraction of the pentosidine precursors accumulated in uremic plasma have a lower than 5000 Da molecular weight. Addition of aminoguanidine and OPB-9195, which inhibit the Maillard reaction, lowered pentosidine yield in both uremic and control plasma. When ultrafiltrated plasma was exposed to 2,4 dinitrophenylhydrazine, the yield of hydrazones, formed by interaction with carbonyl groups, was markedly higher in uremic than in control plasma. These observations strongly suggest that the pentosidine precursors accumulated in uremic plasma are carbonyl compounds. These precursors are unrelated to glucose or ascorbic acid, whose concentration is either normal or lowered in uremic plasma. They are also unrelated to 3-deoxyglucosone, a glucose-derived dicarbonyl compound whose level is raised in uremic plasma: Its addition to normal plasma fails to increase pentosidine yield. This study reports an elevated level of reactive carbonyl compounds ("carbonyl stress") in uremic plasma. Most have a lower than 5000 Da molecular weight and are thus partly removed by hemodialysis. Their effect on pentosidine generation can be inhibited by aminoguanidine or OPB 9195. Carbonyl stress might contribute to AGE modification of proteins and thus to clinically relevant complications of uremia. PMID- 9848793 TI - Role of cytokines in transplantation tolerance: lessons learned from gene knockout mice. PMID- 9848794 TI - Biological basis of hypoalbuminemia in ESRD. AB - Hypoalbuminemia is associated with mortality in patients with end-stage renal disease (ESRD) maintained either on peritoneal dialysis (PD) or hemodialysis (HD). Serum albumin concentration is determined by its rate of synthesis, by the catabolic rate constant (the fraction of the vascular pool catabolized per unit time), by external losses, and by redistribution from the vascular to the extravascular space. Hypoalbuminemia in dialysis patients is primarily a consequence of reduced albumin synthesis rate in both HD and PD patients, and in the case of PD patents, of transperitoneal albumin losses as well. Continuous ambulatory peritoneal dialysis patients are able to increase albumin synthesis to replace losses. Thus, ESRD does not directly suppress albumin synthesis. The rate of albumin synthesis is inversely proportional to the serum concentration of one potential acute phase protein (alpha2 macroglobulin), and albumin concentration is inversely proportional to that of either C-reactive protein or serum amyloid A in both HD and PD patients. The cause of decreased albumin synthesis is primarily a response to inflammation (the acute phase response), although it is possible that inadequate nutrition may also contribute. The cause of the inflammatory response is not immediately evident. There is no evidence that shifts of albumin to the extravascular space or that dilution of the plasma by volume expansion plays any role in causing hypoalbuminemia in ESRD patients. PMID- 9848795 TI - Reflux nephropathy. PMID- 9848796 TI - Prophylactic hemodialysis in the treatment of acute renal failure. Annals of Internal Medicine, 53:992-1016, 1960. AB - Prophylactic hemodialysis has been employed in the treatment of 15 patients with acute renal failure due to acute tubular necrosis (12), bilateral renal cortical necrosis (two), and poststreptococcal glomerulonephritis (one). Dialyses, usually lasting six hours each, were begun before clinical evidence of uremia developed in each patient and/or before the nonprotein nitrogen reached 200 mg.%, and were repeated daily or often enough to maintain the nonprotein nitrogen below 150 mg.%. The hypothesis underlying this technic postulates (1) that wasting, sepsis and impaired wound healing in these patients may reflect tissue injury by the same dialyzable toxic agents which produce the uremic symptoms that are readily reversible by dialysis, and (2) that repeated dialyses should therefore prevent both clinical uremia and the later, often lethal sequelae. The results contrast dramatically with our own past experience in treating patients with acute renal failure with a carefully executed medical regimen together with hemodialysis on conventional indications. Except in one instance of crush injury with progressive intracerebral damage, and one brief occasion in another individual, these patients experienced a stable, convalescent clinical course, remained free of uremic symptoms or chemical imbalances, ate at least three meals daily which were unrestricted in amount and composition, and were ambulatory between dialyses unless confined to bed by associated disease. Wounds healed well. Infection either did not occur, or subsided after appropriate therapy. Fluid restriction was liberalized by means of ultrafiltration with dialysis. Regional heparinization of only the extracorporeal circuit eliminated actual or impending bleeding as a contraindication to dialysis. Chronic vessel cannulation made the frequent dialyses possible, but may have provided the route for repeated, transient bacterial contamination of the blood stream in the first hour of many dialyses. Marked anemia, despite reticulocytosis, moderate to mild weight loss and some mental deficit persisted in spite of the general clinical improvement and well-being. Three patients with tubular necrosis died after seven, 11 and 26 days of oliguria; both patients with bilateral renal cortical necrosis also succumbed, on the seventy-third and ninety-second days of renal failure, and after 29 and 40 dialyses, respectively. At autopsy, evidence of sepsis was conspicuously absent. The remaining 10 patients survived. Thus some, but not all, clinical manifestations of acute renal failure appear to be favorably influenced by prophylactic dialysis treatment. Our initial experience in this group of 15 patients does not of course prove that freedom from complications and a significantly better outlook for survival can be assured to patients with acute renal failure by these methods. However, it seems to offer a reasonable hope of this possibility which we cannot attach to management by medical measures alone, or by dialysis on conventional indications. If this hope is realized in greatly extended, subsequent series, then it seems inevitable that some form of prophylactic dialysis, or some equally effective alternative, should be adopted in treating the majority of patients with acute renal failure. PMID- 9848797 TI - Introduction to the special section: in search of a double paradigm. PMID- 9848798 TI - A longitudinal study of blood pressure in relation to performance on the Wechsler Adult Intelligence Scale. AB - A growth curve analysis was used to examine estimated longitudinal decline (over 19 years) on the Wechsler Adult Intelligence Scale (WAIS; D. Wechsler, 1955) in relation to arterial hypertension and blood pressure (BP) for 55 men and 85 women (40 to 70 years old) who were free from overt major coexisting diseases. BP was associated with longitudinal decline for Visualization-Performance (VP) ability and Speed. Hypertension was a weaker predictor of VP and was unrelated to Speed. Age effects on VP were overestimated when averaged BP level, particularly systolic BP, was not controlled statistically. It was concluded that initial BP level and hypertension predict age-related longitudinal decline in specific major abilities assessed by the WAIS but that BP averaged over examinations is a stronger predictor. PMID- 9848800 TI - Do medical conditions affect cognition in older adults? AB - Analyses of a nationally representative sample who completed a list recall task (weighted n = 6,446) and 2 mental status tasks (weighted n = 6,646) were conducted to determine whether specific medical conditions such as high blood pressure and diabetes as well as general health ratings predict cognitive performance in adults aged 70 to 103. Presence of stroke and poorer health ratings predicted poorer performance on the 3 tasks. Presence of diabetes predicted poorer performance on recall and 1 mental status task. Age interacted with medical conditions including high blood pressure and diabetes in predicting mental status, with condition-related deficits confined to the younger end of the age continuum. Global health ratings interacted with age, with poorer ratings associated with worse mental status in the younger participants. Findings suggest that stroke and diabetes are associated with cognitive deficits. Some deficits are more pronounced in younger old adults with high blood pressure and poorer health ratings. PMID- 9848801 TI - Tailoring information to enhance quitting in smokers with low motivation to quit: three basic efficacy questions. AB - Tailoring information to a target individual's features is a promising line of development in self-help interventions. In this article, 752 smokers with explicit low intention to quit were randomly assigned to 1 of 5 conditions: (a) multiple tailored letters with self-help guide, (b) multiple tailored letters only, (c) a single tailored letter with a self-help guide, (d) a single tailored letter only, or (e) a nontailored intervention. Follow-up assessment took place 4 months after the intervention. Results indicated that the single tailored intervention only had no surplus value compared with a nontailored look-alike intervention. The addition of a self-help guide to a tailored intervention was only useful in highly dependent smokers, and multiple tailoring was more effective than single tailoring. It remains important to elucidate why and for whom certain tailored interventions are more effective. PMID- 9848799 TI - Influence of HIV status and age on cognitive representations of others. AB - In 2 studies the postulate that the perception of time left in life influences the ways that people conceptualize social relationships was explored. It was hypothesized that when time is limited, emotional aspects of relationships are highly salient. In Study 1, a card-sort paradigm involving similarity judgments demonstrated, for a sample of persons 18 to 88 years old, that the prominence of affect in the mental representations of prospective social partners is positively associated with age. In Study 2, the same experimental approach was applied to a sample of young gay men similar to one another in age, but notably different in their health status (that is, HIV negative; HIV positive, asymptomatic; and HIV positive, symptomatic). It was found that, with age held constant, increasing closeness to the end of life is also associated with an increasing prominence of affect in the mental representations of social partners. The results suggest that the perception of limited time, rather than chronological age, is the critical variable influencing mental representations of social partners. PMID- 9848802 TI - Coronary heart disease moderates the relationship of chronic stress with the metabolic syndrome. AB - Metabolic syndrome levels (MSLs) were compared in caregivers (CGs) of spouses with Alzheimer's disease who had diagnoses of coronary heart disease (CHD; n = 27) with non CGs with CHD diagnoses (n = 18), and CGs (n = 44) to non CGs (n = 52) free of CHD. MSLs were greater for CGs than non CGs, but only in persons with CHD (CHD, B for CG status = -.41; non CHD, B = .12; p < .05) at study entry (Time 1 = T1) and CHD, B = -.32; non CHD, B = .14; p < .05) 15-18 months later (Time 2 = T2). In the CHD group, MSLs were associated with poorer health habits at T1 (r = .39, p < .01), uplifts (r = -.37, p < .01) at T2, and CG status (p < .05) at T1 and T2. Relationships of CG status and MSLs declined in the presence of poor health habits at T1 and uplifts at T2. Poorer health habits and fewer uplifts may be associated with elevated MSLs in CGs with CHD. PMID- 9848803 TI - Interpersonal concerns and psychological difficulties of psoriasis patients: effects of disease severity and fear of negative evaluation. AB - Psoriasis creates interpersonal difficulties for many sufferers, but little research has examined factors that contribute to the degree of social and psychological disability that a particular person experiences. In all, 318 psoriasis patients completed measures of psychological and social well-being, the severity of their psoriasis, and their dispositional level of fear of negative evaluation (FNE). Analyses showed that disease severity and FNE significantly predicted perceptions of being stigmatized, interpersonal discomfort, stress over others' reactions, distress regarding the observable symptoms of the disease, the degree to which psoriasis interfered with the patients' lives, and patients' quality of life. Furthermore, FNE exerted a particularly strong influence for patients who had severe cases of psoriasis. PMID- 9848804 TI - Agency, communion, and cardiovascular reactivity during marital interaction. AB - The concepts of agency and communion have been used to describe sex differences in vulnerability to specific stressor domains. This study examined blood pressure and heart rate responses of 60 married couples to experimental manipulations of disagreement (i.e., communion stressor) and achievement challenge (i.e., agency stressor). Consistent with predictions, disagreement elicited heightened cardiovascular reactivity among wives, but not husbands. In contrast, the achievement challenge elicited heightened cardiovascular reactivity among husbands, but not wives. Participants' responses to a circumplex measure of interpersonal appraisal were consistent with the interpretation of differential responses to agency and communion stressors. Results are congruent with a situational approach to sex differences in cardiovascular reactivity and illustrate the utility of interpersonal methods in the explication of psychosocial risk for cardiovascular disease. PMID- 9848805 TI - AIDS treatment advances and behavioral prevention setbacks: preliminary assessment of reduced perceived threat of HIV-AIDS. AB - Recent advances in AIDS treatment have brought renewed optimism for prolonging the lives of those infected with HIV. This article examined beliefs about how new treatments may reduce HIV transmission risk among 298 HIV-negative gay and bisexual men attending a gay pride festival. Results from an anonymous survey showed that men who practiced unprotected anal intercourse as the receptive partner (UAR intercourse) were younger, less well educated, and more likely to believe that it is safe to have UAR intercourse with an HIV-positive man who has an undetectable viral load and that new treatments for HIV relieve their worries about unsafe sex. As HIV treatments continue to advance, new challenges for HIV prevention will likely emerge. PMID- 9848806 TI - Effects of social support and personal coping resources on depressive symptoms: different for various chronic diseases? AB - Effects of psychosocial coping resources on depressive symptoms were examined and compared in older persons with no chronic disease or with recently symptomatic diabetes mellitus, lung disease, cardiac disease, arthritis, or cancer. The 719 persons without diseases reported less depressive symptoms than the chronically ill. Direct favorable effects on depressive symptoms were found for having a partner, having many close relationships, greater feelings of mastery, greater self-efficacy expectations, and high self-esteem. Buffer effects were observed for feelings of mastery, having many diffuse relationships, and receiving emotional support. Buffer effects were differential across diseases for emotional support (in cardiac disease and arthritis only) and for diffuse relationships (in lung disease). Receiving instrumental support was associated with more depressive symptoms, especially in diabetes patients. PMID- 9848807 TI - Measuring self-efficacy in the context of HIV risk reduction: research challenges and recommendations. AB - Self-efficacy theory has had considerable influence in studies of health-related behaviors, including the prevention of sexual transmission of HIV. However, imprecise operationalization of self-efficacy theory in HIV prevention research is common and has important implications for the prediction of risk-reducing behavior from self-efficacy beliefs. In many instances, constructs other than self-efficacy have been assessed. In this article, the operationalization and measurement of self-efficacy in the context of HIV-risk-reduction research are reviewed and challenges inherent to such efforts are identified. Recommendations for enhancing the prediction of risk-reducing behavior from self-efficacy beliefs are also provided. PMID- 9848808 TI - The APOE epsilon4 allele is associated with decline on delayed recall performance in community-dwelling older adults. AB - OBJECTIVE: This study investigated whether the Apolipoprotein (APOE) epsilon4 allele was associated with cognitive decline in community-dwelling older adults. DESIGN: Longitudinal cognitive performance of older adults with the epsilon3/epsilon4 genotype was compared with that of older adults with the epsilon3/epsilon3 genotype. SETTING: Aging Clinical Research Center, Stanford University. PARTICIPANTS: One hundred community-dwelling older adults were recruited from a pool of 531 individuals who had participated in a memory training study 4 to 5 years earlier. These individuals were concerned about their memory functioning and were recruited through newspaper advertisements and contacts with local senior centers. The 100 individuals who agreed to participate in the follow-up investigation were between 59 and 95 years of age. MEASUREMENTS: At both baseline and follow-up, subjects were administered a battery of seven cognitive tests that examined verbal and spatial memory, attention, speed-of processing, and language abilities. APOE genotype was determined at follow-up. RESULTS: Individuals with the epsilon3/epsilon4 APOE genotype were significantly younger than individuals with the APOE epsilon3/epsilon3 genotype. No significant differences were observed between the two groups on measures of attention, speed of-processing, vocabulary, immediate verbal memory, and immediate spatial memory. However, those older adults with the epsilon3/epsilon4 genotype exhibited significantly greater decline in performance on delayed recall of verbal material than did those with the epsilon3/epsilon3 APOE genotype. CONCLUSION: These findings are consistent with previous studies, which suggest that the APOE epsilon4 allele predicts decline on measures of delayed recall. PMID- 9848809 TI - Psychiatric diagnosis and intervention in older and younger patients in a primary care clinic: effect of a screening and diagnostic instrument. AB - OBJECTIVES: To determine whether patient age is associated with psychiatric diagnosis or provider intervention in a busy primary care clinic, and, if so, whether a screening and diagnostic tool, the PRIME-MD, modifies age-related differences. DESIGN, SETTING, AND PARTICIPANTS: PRIME-MD use, psychiatric diagnosis, and provider interventions for psychiatric conditions were recorded for eligible patients attending a Veterans Affairs Medical Center primary care clinic. Data from 952 younger (<65 years) and 1135 older patients (> or =65 years) were analyzed to determine whether there were age-related differences in diagnosis/intervention and if use of the PRIME-MD modified these differences. INTERVENTION: Implementation of the PRIME-MD, a two-step instrument consisting of a self-administered patient questionnaire and a provider-administered structured diagnostic interview. MEASUREMENTS: Outcome measures were rates of (1) PRIME-MD use, (2) overall psychiatric diagnosis, (3) new psychiatric diagnosis, and (4) provider intervention for psychiatric conditions. RESULTS: There was no association between patient age and PRIME-MD use. Older patients were less likely to receive a psychiatric diagnosis in analyses that adjusted for "highly positive" screening questionnaires (OR = .45; P<.001). Older patients were also less likely to receive an intervention for a psychiatric condition in analyses that adjusted for whether a psychiatric diagnosis (OR = .62, P = .015) or a new psychiatric diagnosis (OR = .36, P<.001) was made during the study visit. The PRIME-MD increased rates of diagnosis and intervention but did not alter age related disparities. CONCLUSIONS: Decreased rates of psychiatric diagnosis and intervention in older primary care patients are of concern. Implementing the PRIME-MD will likely increase rates of diagnosis and intervention but will need to be accompanied by additional measures to eliminate age-related disparities. PMID- 9848810 TI - Screening for dementia in the outpatient setting: the time and change test. AB - OBJECTIVE: To develop and validate the Time and Change (T&C) test, a simple, standardized method for detecting dementia in a diverse older outpatient population with varying levels of education. DESIGN: A prospective cohort validation study. SETTING: Two outpatient clinics at an urban teaching hospital. PARTICIPANTS: The concurrent validation sample consisted of 100 consecutive outpatients 70 years of age or older who were 58% non-white and had a 16% dementia prevalence rate and educational levels ranging from 0 to 17+ years. Reliability was tested in a sample of 42 consecutive outpatients 75 years of age or older with a 36% dementia prevalence rate. MEASUREMENTS: T&C ratings were validated against a reference standard based on the Blessed Dementia Rating Scale and the Mini-Mental State Examination. Reliability, contribution to physician recognition of dementia, and ease of use were assessed. RESULTS: In the outpatient setting, the T&C had a sensitivity of 63%, specificity of 96%, a negative predictive value of 93%, a positive predictive value of 77%, and test retest and inter-observer reliability agreement rates of 95% and 100%, respectively. When T&C results were added to the physician's documentation of dementia, the number of missed cases decreased from 44% to 19%, and the number of overcalled cases decreased by 100%. When timed cut points were added, the T&C test had a sensitivity of 94 to 100%, specificity of 37 to 46%, negative predictive value of 98 to 100%, positive predictive value of 23 to 25%, and test retest and inter-observer agreement rates of 82% and 70 to 75%, respectively. CONCLUSION: The T&C test is a simple, accurate, reliable, performance-based tool that can improve physician ability to recognize dementia in diverse outpatient populations. PMID- 9848811 TI - Equivalence of two screens of cognitive functioning: the Short Portable Mental Status Questionnaire and the Orientation-Memory-Concentration test. AB - OBJECTIVE: To determine the equivalence of two screens of cognitive functioning: the Short Portable Mental Status Questionnaire (SPMSQ) and the Orientation-Memory Concentration (OMC) test. DESIGN: The design was cross-sectional and longitudinal. SETTING: Four rural and one urban county in the Piedmont region of North Carolina (n = 3210). PARTICIPANTS: A stratified random cluster sample (n = 3210) of people 68 years of age and older. MEASUREMENTS: SPMSQ and OMC at the fourth wave of the Duke Established Populations for Epidemiologic Studies of the Elderly (EPESE); disability, depression, and death measurements from the fourth through seventh waves of EPESE; demographic characteristics. RESULTS: On the SPMSQ and the OMC (r = .80), 15.3% and 38.4%, respectively, of those tested were rated cognitively impaired. Poorer scores were associated with older age, black race, and less education. These associations were attenuated on the dichotomized SPMSQ but not on the OMC. Both measures predicted disability and depressive symptomatology currently and 3 years hence and death. CONCLUSIONS: The SPMSQ and OMC, although highly correlated, are not equivalent. Association with race and education are greater for the OMC, whereas an association with age exists for both measures. The milder level of impairment identified by the OMC increases predictive capacity. PMID- 9848812 TI - Executive dyscontrol: an important factor affecting the level of care received by older retirees. AB - OBJECTIVES: To examine the relative contributions of Executive Control Function (ECF), general cognition, mood, problem behavior, physical disability, demographic variables, and the number of prescribed medications to the level of care received by older retirees. DESIGN: Multivariate regression and discriminant modeling. SETTING: A single Continuing Care Retirement Community (CCRC) in San Antonio, Texas. PARTICIPANTS: A total of 107 older retirees (mean age = 83.7+/ 7.2 years), including 17 community-dwelling, well, older controls and 90 CCRC residents. CCRC subjects represented a convenience sample of consecutive referrals for geropsychiatric assessment. Sixty-one subjects resided at a noninstitutionalized level of care, and 46 were institutionalized. MEASUREMENTS: Tests of ECF (the Executive Interview (EXIT25)), general cognition (the Mini Mental State Examination (MMSE)), mood (the Geriatric Depression Scale short-form (sGDS)), problem behavior (the Nursing Home Behavior Problem Scale (NHBPS)), physical disability (the Cumulative Illness Rating Scale (CIRS)), age, gender, years of education, and the number of prescribed medications were studied. RESULTS: All variables except gender and education varied significantly across level of care. Four variables made significant independent contributions; EXIT25 score (r2 = .48, P< .001), medication usage (partial r2 = .11, P<.001), sGDS score (partial r2 = .06, P = .001), and problem behavior (partial NHBPS r2 = .04, P<.04). These variables accounted for 69% of the total variance in level of care (R2 = .69; F (df 7,99) = 32.1, P<.001). A discriminant model based on the number of prescribed medications, EXIT25, sGDS, and NHBPS scores classified 83.2% of cases correctly (Wilke's lambda = .50, F(5,101) = 20.1; P<.001). The MMSE enters but fails to contribute significantly, independent of the other variables. Age and CIRS scores fail to enter. CONCLUSIONS: Cognitive (particularly ECF) impairment contributes most to the observed variance in level of care received by older retirees in this CCRC. In contrast, markers of general cognition, depression, and physical illness contributed relatively little additional variance. ECF is not detected well by traditional cognitive measures and must be sought by specific tests. Further study is needed to replicate these findings in other populations. PMID- 9848813 TI - Functional outcomes of posthospital care for stroke and hip fracture patients under medicare. AB - BACKGROUND: Medicare's introduction of the Prospective Payment System for hospitals has led to tremendous growth in ways of providing posthospital care. Despite substantial differences in costs per episode of care, the type of posthospital care that produces the best results for specific types of patients is not clear. This study analyzed the outcomes of different types of posthospital care for a cohort of older Medicare patients (who had diagnoses associated with the use of a range of posthospital care modalities) for up to a year after hospital discharge. METHODS: Medicare patients hospitalized with strokes and hip fractures were enrolled consecutively just before discharge from 52 hospitals in three cities in 1988-1989. These diagnosis-related groups were chosen because patients were discharged to all three major types of Medicare-supported posthospital care. Patients were interviewed in-person before discharge and again at 6 weeks, 6 months, and 1 year after discharge. The functional outcomes of posthospital care were evaluated by the instrumental variables estimation approach to correct for selection bias caused by nonrandom treatment assignment. The impacts of discharge locations on the functional outcomes were examined by one-way analyses of variance (ANOVA). RESULTS: In general, the more disabled patients went to nursing homes and rehabilitation, but the overlap in distribution was sufficient to conduct the analyses. Stroke patients discharged to nursing homes had the highest mortality rate (P<.01). Stroke patients discharged to home health had the lowest rehospitalization rates (P<.05). Hip fracture discharged to home health care had the highest adjusted rehospitalization rate, whereas those discharged to nursing homes had the lowest adjusted rehospitalization rate (P<.05). For stroke patients, posthospital care in rehabilitation facilities or home health care was associated with significantly better functional improvement compared with stroke patients discharged elsewhere. However, functional outcomes deteriorated by 1 year posthospitalization among stroke patients who received their posthospital care at nursing homes or received no formal posthospital care. For hip fracture patients, all four types of posthospital care were associated with functional improvement, but patients discharged to rehabilitation facilities experienced the most functional improvement. CONCLUSIONS: The choice of posthospital care can influence the course of Medicare patients. Careful attention should be paid to how hospital discharge decisions are made and to the financial incentives for different types of posthospital care provided under the current payment system. The current supply of nursing homes is not well suited to the demands of posthospital care. PMID- 9848814 TI - Aging, esophageal motility, and gastroesophageal reflux. AB - OBJECTIVES: To compare esophageal motility and gastroesophageal reflux characteristics in young, middle-aged, and older healthy volunteers. DESIGN: Comparison of conventional esophageal manometry and scintigraphic study of gastroesophageal reflux in volunteers aged 20 to 30 years, 50 to 60 years, and 70 to 80 years. SETTING: Gastroenterology and nuclear medicine laboratories of a tertiary care university hospital. PARTICIPANTS: Forty healthy adult volunteers recruited from the community, 20 aged 20 to 30 years, 10 aged 50 to 60 years, and 10 aged from 70 to 80 years. MEASUREMENTS: Each volunteer underwent conventional esophageal manometry and scintigraphic study of gastroesophageal reflux. Contractile wave amplitude, duration, velocity, and lower esophageal sphincter relaxation duration, as well as the presence of abnormal peristalsis, were correlated with the proportion of volunteers with gastroesophageal reflux and the number and duration of gastroesophageal reflux episodes. RESULTS: Quantitative manometric parameters showed no correlation with gastroesophageal reflux patterns. Abnormal peristalsis was found more frequently in the older volunteers. The number of gastroesophageal reflux episodes per volunteer was similar in the three age groups, but the duration of gastroesophageal reflux episodes was longer in the older volunteers. CONCLUSION: Healthy older persons have impaired clearance of refluxed materials associated with a high incidence of defective esophageal peristalsis. This may explain the higher severity of reflux esophagitis in older people. PMID- 9848815 TI - Validation and application of the pneumonia prognosis index to nursing home residents with pneumonia. AB - OBJECTIVES: To evaluate the predictability of a pneumonia prognosis index in nursing home residents with pneumonia and to use the index to account for acute severity of pneumonia before comparing the short-term outcome of residents with pneumonia treated with intravenous antibiotic therapy in two different settings: an inpatient geriatrics unit and a nursing home DESIGN: A retrospective chart review of 158 episodes of nursing home-acquired pneumonia treated initially with intravenous antibiotics; 100 episodes were treated in an inpatient acute geriatrics service (AGS), and 58 were treated completely in a nursing home (Nursing Home group) SETTING: The AGS is a 20-bed unit within a 400-bed, public, university-affiliated hospital. The Nursing Home group consisted of residents of two nonproprietary nursing homes. PARTICIPANTS: Nursing home residents with radiographically proven pneumonia who had at least one of the following signs/symptoms: cough, fever, purulent sputum, respiratory rate > or = 25 per minute, localized auscultatory findings, or pleuritic pain. MEASUREMENTS: The pneumonia prognosis index was calculated for each resident at the time of diagnosis of pneumonia; the index has been validated as a predictor of hospital outcome in patients with community-acquired pneumonia and is also considered a measure of acute severity of pneumonia. Status (alive or dead) of each resident at 30 days after diagnosis was the major dependent variable RESULTS: Mean (+/-SD) duration of antibiotic therapy for the Nursing Home group (10.7+/-4.5 days) was not significantly different from that of the AGS group (9.6+/-3.4 days; P = .26). The pneumonia prognosis index stratified the 158 episodes of pneumonia into low- and high-risk groups for 30-day mortality; the mortality rates in each risk strata were not significantly different from those reported in the original derivation and validation studies of the index. In addition, the distribution of episodes among the risk strata of the index was not significantly different for the two study groups, which was an indication that the two groups were similar in terms of acute severity of pneumonia. Thirty-day mortality was not significantly different between the two groups: AGS, 21% and Nursing Home, 24.1% (P = .66). CONCLUSION: The pneumonia prognosis index seems to have the same capability for predicting the outcome in nursing home residents with pneumonia as in residents with community-acquired pneumonia. The index is also a measure of acute pneumonia severity. Nursing home residents with pneumonia, even those who are most acutely ill, can be treated successfully with intravenous therapy in the nursing home; their 30-day mortality was no different than that of those with the same acute severity of illness who were admitted to a hospital for treatment. PMID- 9848816 TI - Venous thrombosis in older people: prevalence of the factor V gene mutation Q506. AB - OBJECTIVES: Old age is usually considered to be a risk factor for venous thromboembolism, in conjunction with other factors such as heart failure, major surgery, cancer, long-term immobilization, and antiphospholipid antibodies. Genetic risk factors, especially inherited deficiencies in coagulation inhibitors, also play a role in the pathogenesis of thrombosis, but these are usually diagnosed in thrombophilic patients before the age of 50. The factor V Q506 mutation, responsible for activated protein C resistance, was recently linked to thromboembolic disease. We therefore investigated the prevalence of biological risk factors in older hospital patients with venous thromboembolism. DESIGN: A 2-year study period. SETTING: Ivry sur Seine (Paris), France. PARTICIPANTS: Seventy-nine geriatric patients (60 women and 19 men, mean age 83+/ 6.8 years, range 70-102 years) who had had at least one proven episode of venous thromboembolism were enrolled over a 2-year period. MEASUREMENTS: Lupus anticoagulant and antithrombin (AT), protein C (PC), and protein S (PS) levels were determined in plasma. The factor V Q506 mutation was detected on genomic DNA. RESULTS: Lupus anticoagulant was detected in two women, one of whom also had a high level of anticardiolipin IgG, leading to the diagnosis of an antiphospholipid syndrome. No hereditary deficiency in AT, PC, or PS was found, but one patient had an acquired AT deficiency. Interestingly, nine of the 79 patients (11.4%, six women and three men) were heterozygous for the factor V Q506 mutation, although none were homozygous. The only major risk factor for thrombosis identified in these patients was prolonged immobilization in four cases. Four of the nine patients who were heterozygous for the factor V Q506 mutation had recurrent thromboembolism, and two of these patients had been immobilized for long periods. CONCLUSIONS: This study confirms that hereditary deficiencies in coagulation inhibitors, and the lupus anticoagulant, are rarely involved in the pathogenesis of venous thromboembolism in older subjects. In contrast, the factor V Q506 mutation was frequently associated with thrombosis (11.4% of our patients) and should, therefore, be considered an important risk factor in the older people. PMID- 9848817 TI - Evaluating the use of angiotensin-converting enzyme inhibitors for older nursing home residents with chronic heart failure. AB - BACKGROUND: Despite their well noted therapeutic benefits for heart failure (HF), angiotensin-converting enzyme (ACE) inhibitors may be underprescribed and underdosed among older nursing home patients. OBJECTIVES: To assess the use of ACE inhibitor therapy in older (> or =70 years) nursing home residents with systolic heart failure (HF). DESIGN: A cross-sectional, retrospective analysis. SETTING: Five long-term care facilities in Providence, Rhode Island. SUBJECTS: Adults aged 70 years or older with left ventricular (LV) ejection fractions < or =40%. MEASUREMENTS: New York Heart Association (NYHA) class, comorbid diseases, and cardiac medications with logistic regression analysis to clarify their bearing on the prescription of ACE inhibitors. RESULTS: Of the 819 nursing home residents who were evaluated, 119 (24 men, 95 women) fulfilled exacting entry criteria, i.e., heart failure signs/symptoms and documented LV systolic dysfunction. Forty-one of these 119 (35%) older persons were receiving ACE inhibitor therapy, predominantly in doses (< or =50 mg captopril/day or < or =5 mg enalapril/ day) less than those of proven therapeutic efficacy. Compared with older residents not receiving ACE inhibitors, those receiving ACE inhibitors included fewer with NYHA Class I HF (0 vs 21%, P = .017), more men (58 vs 28%, P< or =.01), and more people with hypertension (61 vs 35%, P< or =.01). In contrast, diuretics were prescribed more frequently among those not receiving ACE inhibitors (83 vs 56%, P< or =.001). CONCLUSIONS: ACE inhibitors are underprescribed and underdosed among elderly nursing home patients carefully screened to include systolic HF and no contraindications to the medication. ACE inhibitors are particularly underused in those elderly with NYHA Class I HF and in those receiving diuretics. PMID- 9848818 TI - Prevalence and outcome of methicillin-resistant Staphylococcus aureus colonization in two nursing centers in Georgia. AB - OBJECTIVES: To evaluate the cost-effectiveness and validity of aggressive diagnosing and eradication of methicillin-resistant Staphylococcus aureus (MRSA) colonization. DESIGN: An observational study of MRSA colonization. SETTING: Two skilled nursing facilities in Georgia. PARTICIPANTS: Eighty-seven residents of the above-mentioned skilled nursing facilities INTERVENTION: No changes were made in either nursing facility practices or patient treatments. MEASUREMENTS: The presence or absence of MRSA colonization was determined at the beginning of the study, after hospitalizations or absences from the facility, and 12 months after initial testing. RESULTS: Eighty-seven participants began the study, and 56 completed it. At the beginning of the study, 17.9% of the 56 who later completed the study were positive for MRSA. By the end of the study, colonization had decreased to 14.3%. The difference in the number of positive cultures for residents returning from the hospital compared with those returning from non hospital settings was not statistically significant (P = .546). No deaths or hospitalizations were attributable to MRSA infection. CONCLUSION: Because no significant correlation was found between MRSA colonization and active MRSA related infections, patients colonized by MRSA should not be barred from admission to nursing homes. PMID- 9848819 TI - Gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) is a spectrum of disease that can best be defined as the symptoms and/or signs of esophageal or adjacent organ injury secondary to the reflux of gastric contents into the esophagus or, beyond, into the oral cavity or airways. Injury is defined based on symptoms or organ damage resulting in esophagitis, laryngeal inflammation, or acute and/or chronic pulmonary injury. Though specific studies are lacking, GERD seems to occur more frequently in older people. In general, because it has been present for a longer period of time, older patients will often present with more complicated GERD, including severe erosive esophagitis, peptic stricture, and Barrett's esophagus, a premalignant condition, making treatment of older patients with GERD potentially more difficult and complex. Though it is not usually life threatening, GERD can have a major effect on patients' well-being and quality of life. This article reviews the presentation, pathophysiology, diagnosis, and treatment of GERD as it relates to the older patient. PMID- 9848820 TI - Current status of antioxidant therapy for Alzheimer's Disease. AB - Accumulating evidence from preclinical and clinical studies supports the hypothesis that oxidative stress may be associated with the onset and progression of Alzheimer's Disease (AD). Antioxidant therapies are being promoted in the lay press to enhance mental functions and delay cognitive losses with aging. An increasing number of physicians are also recommending antioxidant therapies, such as high dose vitamin E, for subjects with AD and other neurodegenerative disorders. High dose vitamin E, ginkgo biloba, and selegiline are three putative antioxidants that have been tested in randomized multicenter trial conditions in the US. This paper summarizes the oxidative stress hypothesis of AD and reviews the strengths and limitations of published antioxidant studies in AD in relation to the role of such therapies in practice. PMID- 9848821 TI - What should we do about undertreatment of late life psychiatric disorders in primary care? PMID- 9848822 TI - Recognition of dementia: discovering the silent epidemic. PMID- 9848823 TI - Problems in measuring and interpreting cognitive decline. PMID- 9848824 TI - Cognitive status and mortality in community-residing older people. PMID- 9848825 TI - Alendronate--risk for esophageal stricture. PMID- 9848826 TI - Sertraline-induced hyponatremia in an older patient. PMID- 9848827 TI - Long-term ambulatory monitoring of bladder temperature in older people: a pilot study. PMID- 9848828 TI - Investigation of polymyositis-dermatomyositis in older people should include rectal examination. PMID- 9848829 TI - Is O-K okay? PMID- 9848830 TI - Identification and enumeration of staphylococci from the eye during soft contact lens wear. PMID- 9848831 TI - Mooren's ulcer. AB - BACKGROUND: Mooren's ulcer is a rapidly progressive, painful, ulcerative keratitis which initially affects the peripheral cornea and may spread circumferentially and then centrally. Mooren's ulcer can only be diagnosed in the absence of an infectious or systemic cause and must be differentiated from other corneal abnormalities, such as Terrien's degeneration. Although the etiology remains unknown, recent research has proposed an underlying immune process and a possible association with the hepatitis C virus. The response to medical and surgical intervention is typically poor, and the visual outcome can be devastating. CASE SERIES: Three patients presented to our clinic with inferior peripheral corneal defects characteristic of Mooren's ulceration. The first patient, a 67-year-old white male, presented with an area of progressive peripheral thinning of the left inferior cornea 1 week after a preoperative skin cleanser was inadvertently splashed in both eyes. This occurred during a surgical procedure to remove a basal cell carcinoma. The second patient, a 56-year-old white male, was treated for a recurrent left inferior corneal ulcer with impending risk of perforation. The third patient was a 68-year-old white male referred for a painful left inferior peripheral ulcer, which rapidly progressed into a bilateral corneal melt disorder. All patients were diagnosed with Mooren's ulcerative keratitis after they underwent extensive medical and laboratory testing to rule out an infectious or systemic cause of their corneal melt. The first patient was treated with oral steroids, as well as doxycycline, to control his acne rosacea. The second patient responded to aggressive treatment with topical steroid therapy. This patient also tested positive for hepatitis C. The third patient rapidly developed a perforated left cornea and was treated with a penetrating keratoplasty after a patch graft had failed. DISCUSSION: Mooren's ulcer is an idiopathic, painful, progressive ulceration of the peripheral cornea. These ulcers usually respond poorly to conventional therapy, as there is limited knowledge of the pathophysiology of the disease. Evidence of an autoimmune component advocates the use of steroids and immunosuppressive agents. With further research and understanding of Mooren's ulcer, better treatment options may be available in the future. PMID- 9848832 TI - Corneal response to orthokeratology. AB - PURPOSE: The technique of orthokeratology produces a corneal response to the mechanical pressures exerted by rigid contact lenses. This paper reports a study which investigated the topographic and pachometric corneal changes induced by orthokeratology. METHODS: Six young myopic subjects (11 eyes) wore "accelerated orthokeratology" lenses (OK-74; Contex Inc., Sherman Oaks, CA) in a high Dk material (AirPerm; Dk = 88) for 28 days. Corneal and epithelial thickness were measured topographically using the Holden-Payor optical micropachometer, and corneal topography was monitored using the EyeSys system. RESULTS: Refractive error change reached 1.71 +/- 0.59 D reduction in myopia after 28 days. After 1 day of lens wear, statistically significant central corneal flattening was noted, which progressed to reach 0.22 +/- 0.07 mm (1.19 +/- 0.38 D) at 28 days. A trend toward central epithelial thinning was apparent, reaching statistical significance on day 28 (7.1 +/- 7.1 microm; 9.6%). Midperipheral corneal thickening was also found approximately 2.5 mm from the corneal center, which was statistically significant by day 14 (13.0 +/- 11.1 microm; 2.4%). Calculations using Munnerlyn's formula indicate that changes in corneal sagittal height based on topographical thickness changes across the flattened central 5.25-mm zone can account for the refractive changes observed. CONCLUSIONS: These findings suggest that the initial corneal response to orthokeratology may be explained by redistribution of corneal tissue, rather than by overall bending of the cornea. PMID- 9848833 TI - Effect of topical anesthetic use on initial patient satisfaction and overall success with rigid gas permeable contact lenses. AB - BACKGROUND: Rigid gas permeable (RGP) contact lenses have numerous benefits; however, RGP lens use is not increasing in the United States. An important factor for this trend has been initial comfort. Studies have demonstrated that how RGPs are presented to patients, in addition to lens design, can play an important role in the initial comfort process. Another important factor could be the use of a topical anesthetic during the fitting and dispensing visits. The purpose of this study was to use a multicenter format to determine if topical anesthetic use increased the likelihood of patient satisfaction and success. METHODS: A total of 80 subjects, with no previous rigid lens wear experience, was entered into this 1 month study, including 20 subjects from each of 4 institutions. Subjects were randomly divided into the following two groups: (A) anesthetic or (B) placebo, with the former group receiving one drop of a topical anesthetic before lens insertion at both the diagnostic fitting and dispensing visits, whereas the latter group received a placebo. Subjects completed a questionnaire on their perception of rigid lens wear both immediately before fitting and at the 1-month visit. After diagnostic fitting with rigid lenses, subjects completed an adaptation questionnaire after 15 min, 1 week, 2 weeks, and 1 month of lens wear. RESULTS: Seventy of the 80 subjects completed the study and, of the 10 subjects who discontinued, 8 were in the placebo group. In all categories evaluated, the anesthetic group experienced a more optimum adaptation experience at each visit vs. the placebo group. Specifically, overall comfort was rated significantly higher at both dispensing and 2 weeks. In addition, the anesthetic group exhibited significantly greater overall satisfaction with rigid lens wear at 2 and 4 weeks. Also, the anesthetic group perceived their adaptation, sensitivity, and adaptation time to be significantly better at the 1-month visit. There was no significant difference in corneal staining between these two groups at each visit, with the exception of a greater amount of staining in the central quadrant for the placebo group at the 1-month visit. CONCLUSIONS: The use of a topical anesthetic at the fitting and dispensing visits for first-time wearers of RGP lenses resulted in significantly fewer dropouts, improved initial comfort, an enhanced perception of the adaptation process, and greater overall satisfaction after 1 month of lens wear as compared to the use of a nonanesthetizing placebo at those visits. This result, in combination with both presenting RGP lenses in a nonthreatening manner and optimizing the lens design and fitting relationship, should result in a positive adaptation process and successful wear of RGP contact lenses. PMID- 9848834 TI - Development and validation of a visual acuity chart for Australian Aborigines and Torres Strait Islanders. AB - BACKGROUND: A new visual acuity chart was designed for use with Australia's indigenous population to overcome perceived inadequacies of conventional English letter charts for this group. This chart, which incorporates a black and white turtle icon, is described, and validation data are presented. METHODS: The chart is based on logarithm of the minimum angle of resolution (logMAR) principles and incorporates a turtle symbol modified from the design of an indigenous artist. The task is one of discrimination, with subjects being required to distinguish the split tail of the turtle from its head, which has the same overall shape and average luminance; the body of the turtle provides no directional cues which might assist in this judgment. The chart was validated in two ways: Experiment I. Performance was compared with the Bailey-Lovie and Konig bar charts in terms of unaided visual acuity data for 90 subjects (mean age: 38.3 +/- 20.3 years) and Experiment II. Data were obtained for 10 young subjects for these 3 charts and an Illiterate E chart, with refractive blur imposed with trial lenses over habitual distance corrections (spherical: +0.50, +1.00, +2.00, and +4.00 D; cylindrical: +1.00 and +2.00 D, axes 45, 90, and 180 degrees). To avoid cultural and literacy issues as possible sources of differences in performance between the charts in this validation study, subjects were selected from the wider Australian population rather than specifically from its indigenous segment. RESULTS: Experiment I: The Turtle chart performed most like the Konig Bar chart for this component of the validation exercise. Nonetheless, results for the Turtle chart correlated highly with those for the Bailey-Lovie chart as well as the Konig Bar chart, although there were subtle differences between charts in the rate of decline of visual acuity as visual performance decreased. Experiment II: The turtle chart behaved most like the Illiterate E chart with imposed spherical focusing errors, with the Bailey-Lovie chart showing a faster decline and the Konig Bar chart showing a slower decline in performance, with increasing defocus. All 4 charts showed similar directional biases with astigmatic defocus, being most affected by oblique (45 degrees) astigmatism. CONCLUSION: The Turtle chart met the criteria set for its validation as a visual acuity chart in that it gave comparable results to the other commonly used visual acuity charts, both in the case of unaided vision and when refractive blur was imposed. PMID- 9848835 TI - Inter-clinician agreement on clinical data abstracted from patients' medical charts. AB - PURPOSE: The purpose of this study was to determine the inter-observer repeatability of retrospective chart reviews. METHODS: Retrospective abstraction of corneal transplant patients' medical records by two independent observers was performed at the University of California, Davis, Department of Ophthalmology, Sacramento, CA. The sample consisted of 100 consecutive patients who underwent penetrating keratoplasty, and the primary outcome measure was the degree of agreement between the 2 independent observers on clinical ocular data abstracted from patients' medical charts. RESULTS: Even when given specific criteria for abstraction of data from patients' medical charts, inexperienced but trained observers exhibited a small degree of disagreement (range 0 to 12%) on patient demographic variables (e.g., gender, age, and date of birth) to be used for data analysis. Larger amounts of disagreement (range 3 to 26%) were found on frequently occurring variables (e.g., predisposing ocular conditions, surgical complications, and classification of corneal staining). CONCLUSIONS: Based on these data, inter-observer repeatability for retrospective chart review studies is good, but investigators using this study methodology must be quite specific about the data to be collected and should be aware of standardization techniques to enhance observer repeatability. PMID- 9848836 TI - Assessment of vergence facility in a sample of older adults with presbyopia. AB - PURPOSE: The aim of this study was to establish whether assessment of vergence facility is clinically useful in older adults with presbyopia and to determine pilot normative data for this age range. METHODS: Vergence facility was measured in a sample of 50 asymptomatic subjects with a mean age of 58.7 years using 8delta base-in (BI) and 12delta base-out (BO) prisms mounted in clinical flipper frames. Testing was performed for 1 min on four separate occasions over the course of a few weeks. RESULTS: Of the 42 subjects that completed the study, vergence facility could be satisfactorily measured on only 21 subjects. The mean vergence facility for this group was approximately 7 cycles per minute (cpm), commensurate with previous results in younger subjects using a similar paradigm. There were no differences in vergence facility between the first and second 30-s periods of testing, nor were there any differences in average vergence facility over the 4 test occasions. Test-retest repeatability, however, was poor. The BI prism phase took longer, on average, to fuse than the BO prism phase (BI 5.2 s and BO 3.7 s). CONCLUSIONS: Our results show a high degree of variability in vergence facility in older subjects with presbyopia. More research is required before deciding whether measurements of vergence facility are of use in investigating binocular vision of older patients with presbyopia. PMID- 9848837 TI - Sensory fusion under dynamic background. AB - BACKGROUND: The influence of fixation target and background on the sensory fusion status was studied. METHODS: We examined the sensory fusion of eight normal subjects with a phase difference haploscope (PDH) under different background conditions: no background, a static background, and a dynamic background. Four sizes of a pair of similar pictures (faces) with four different kinds of suppression targets (nose, hat, mouth, and necktie) were used for test targets. During the 1-min fixation of these targets with the different backgrounds, we examined the location and the length of time the suppression targets disappeared. RESULTS: The suppression targets of the small fixation target disappeared easily, compared with those of the large one. The duration of disappearance of suppression targets increased when the dynamic background was used. CONCLUSIONS: The small fixation target and the dynamic background exert a considerable influence on the state of sensory fusion. PMID- 9848838 TI - Modeling human eye aberrations and their compensation for high-resolution retinal imaging. AB - We introduced a mathematical eye model using Gullstrand's six-surface eye model modified by clinically measured aspherical data to study human eye aberrations and their compensation for high-resolution retinal imaging. Ray tracing was used to characterize aberrations and point spread functions (PSFs) of the eye model. By using the Zernike polynomial decomposition of the calculated pupil function, we quantified the wavefront aberrations. Based on calculated PSFs, we designed optical inverse filters to reduce the aberrations for a large pupil size and improve the resolution. Spherical aberration and oblique astigmatism were found to be in good agreement with published experimental measurements. Spherical aberration and defocus were the most significant aberrations for on-axis imaging, whereas oblique astigmatism and coma combined with spherical aberration and defocus were most significant for off-axis imaging. The best retinal image resolution occurred at 2- to 3-mm pupil diameter. After aberration correction for an 8-mm diameter pupil, the resolutions for on-axis or 9 degrees off-axis imaging points were very close to diffraction-limited resolutions. Over a limited field of view (FOV), retinal image resolution of the eye model can be greatly improved by aberration correction using aspheric and astigmatic lenses. For imaging large FOVs, space-variant compensation techniques will be required for aberration correction. PMID- 9848839 TI - First- and third-order optical theory of gradient index materials, with application to contact lenses. AB - PURPOSE: To investigate the feasibility of using gradient index media in contact lenses, we developed simple methods which we used to derive the power and aberrations associated with the contact lenses. METHODS: In one method, we assume that the height of a ray does not change as it passes through the lens. We describe a second method in which the ray is assumed to follow a parabolic path as it passes through the lens. We use sophisticated third-order theory and finite raytracing for comparison with these methods. RESULTS: The methods are compared for contact lenses with parabolic radial gradient index media. Without the gradient index, these lenses would have zero power. The formula for power which assumes no change in ray height gives errors of approximally 0.8 and 1.5% for lenses of thicknesses 0.18 and 0.36 mm. However, the formula for third-order spherical aberration which uses the same assumption gives poor estimations. The method for calculating power using the parabolic ray path is very accurate. The sophisticated third-order aberration theory was reasonably accurate out to 2.5 mm ray height. The contact lenses with the gradient index media have much smaller aberration in air than do conventional contact lenses of the same powers, with the sign of the aberration being reversed. CONCLUSIONS: Our simple procedures give good estimations of powers of contact lenses with gradient index media. The approximate method, which assumes that the height of a ray does not change as it passes through the lens, should not be used for finding the spherical aberration of such a lens. Contact lenses with gradient index media have potential for minimizing spherical aberration. PMID- 9848840 TI - An institutional experience with cervical vagus nerve trunk stimulation for medically refractory epilepsy: rationale, technique, and outcome. AB - OBJECTIVE: Intermittent stimulation of the left cervical vagus nerve trunk is emerging as a novel adjunct in the treatment of medically refractory seizures. We sought to evaluate theoretical and practical issues attendant to this concept. We review the anatomic and physiological background arguing for clinical application of vagus nerve stimulation, discuss salient aspects of patient selection and the nuances of surgical technique, and present our observations of and results from application of the method. METHODS: Each of 18 patients with medically refractory epilepsy and at least six complex partial or secondarily generalized seizures per month underwent placement of a NeuroCybernetic Prosthesis pulse generator (Cyberonics, Webster, TX) in the chest, connected to helical platinum leads applied to the left cervical vagus nerve trunk. The patients were then randomized in a double-blinded fashion to receive either high (presumably therapeutic) or low (presumably less therapeutic) levels of vagus nerve stimulation. Reduction in seizure frequency, global assessments of quality of life, physiological measurements, and adverse events were recorded during a 3-month period. Patients in the low group were then crossed over to high-stimulation paradigms during a 15 month extension trial. RESULTS: All operations were successful, uneventful, and without adverse postoperative sequelae. One patient was excluded from analysis because of inadequate seizure calendars. Of the seven patients initially assigned to high stimulation, the mean reduction in seizure frequency was 71% at 3 months and 81% at 18 months. Five (72%) of these patients had a greater than 75% reduction in seizure frequency, and one (14%) remained seizure-free after more than 1.5 years of follow-up. The mean reduction in seizure frequency among the low-stimulation group was only 6% at 3 months. No serious complications, device failures, or physiological perturbations occurred. CONCLUSION: In our experience, vagus nerve stimulation has proven to be a safe, feasible, and potentially effective method of reducing seizures in select patient populations. However, the elements of strict definition for the application of the method require further study. PMID- 9848841 TI - Selection of cerebral aneurysms for treatment using Guglielmi detachable coils: the preliminary University of Illinois at Chicago experience. AB - OBJECTIVE: We present our initial experience with Guglielmi detachable coils (GDCs). The aim of this study was to determine the criteria for aneurysms, ruptured or unruptured, that are suitable for this technique. The importance of aneurysm geometry and its impact on the final results are discussed. METHODS: A retrospective analysis of 329 patients with 339 cerebral aneurysms that were treated at the University of Illinois Hospital at Chicago from May 1994 to June 1997 was conducted. One hundred eighty-five patients were treated surgically, and 144 were selected for treatment using GDCs. Of the 144 patients selected for GDC treatment, 55 patients with 55 aneurysms were admitted during the acute phase of subarachnoid hemorrhage and 89 patients with 97 aneurysms had nonruptured aneurysms or were treated after clinical recovery of previously ruptured aneurysms. All procedures were performed with the patients under general anesthesia and with systemic heparinization using live simultaneous biplane roadmapping, with the exception of the first four patients. These patients were treated before the installation of the biplane system. The percentage of aneurysm occlusion was determined at the end of each procedure. Follow-up angiography was scheduled to be performed at 6 months, 1 year, and 2 years after treatment. PATIENT SELECTION: For the initial 25 patients (Group 1), selection for coiling was restricted to nonsurgical candidates or patients in whom coiling was thought to be the best treatment choice, based on medical condition and location of the aneurysm. The geometry of the aneurysm was not considered to be an important factor in the selection for coiling. The remaining patients (Group 2) were selected for coiling based on aneurysm geometry, as determined by pretherapeutic angiography. Aneurysms that were considered to be favorable for coiling included those that had a dome-to-neck ratio of at least 2 and an absolute neck diameter less than 5 mm. RESULTS: The initial 25 patients (Group 1) were treated from May 1994 to February 1995. There were high morbidity and mortality rates, with 56% of the treated aneurysms occluded at 6 months. The remaining patients (Group 2) consisted of 119 patients with 123 aneurysms. There was no mortality directly related to the coiling procedure, and permanent morbidity was limited to 1.0%. Three patients (2.5%) developed transient neurological deficits secondary to the procedure, and seven patients (5.8%) experienced periprocedural complications that did not result in neurological sequelae. The morphological results were strongly correlated to the geometry of the aneurysms, with a complete occlusion rate of 72% among the acutely ruptured aneurysms and 80% among the nonacute aneurysms, when patients were selected for treatment based on the geometry of the aneurysms and the dome-to-neck ratio was at least 2. The occlusion rate dropped to 53% when selection was not based on aneurysm geometry and the dome-to-neck ratio was less than 2. A summary of the morphological outcomes for the Group 2 patients shows that 86% of the aneurysms that initially underwent coiling using GDCs were completely occluded (78% by coils alone, 3.0% in conjunction with surgery, and 5.0% with parent artery occlusion). Residual small neck remnants were present in 11% of the Group 2 aneurysms (3.0% were scheduled for surgical treatment of residual neck remnant growths not amenable to further endovascular treatment, and 8% were scheduled for initial 6-mo follow-up examinations). Death resulting from unrelated causes before initial follow-up occurred in 3.0% of the patients. CONCLUSION: These preliminary results suggest that using GDCs is a safe technique resulting in low morbidity and mortality rates for the treatment of intracranial aneurysms in appropriately selected patients. The percentage of complete aneurysm occlusion is related to the density of coil packing, which is strongly dependent on the geometry of the aneurysm. Optim PMID- 9848842 TI - Embolization of neurosurgical lesions involving the ophthalmic artery. AB - OBJECTIVE: A number of anteriorly located cranial base and extracranial lesions receive their vascular supply wholly or in part from the ophthalmic artery, and embolization of the ophthalmic artery can be helpful in the management of these lesions, either as the primary treatment or as an adjunct to surgery. We present situations in which the embolization of lesions involving the ophthalmic artery was performed to effect a partial or total cure of the lesion. METHODS: Twelve patients underwent a total of 15 embolization attempts on lesions involving the ophthalmic artery. Four patients had arteriovenous malformations of the orbit, four had dural arteriovenous fistulae, two had orbital meningiomas, one had a planum sphenoidale meningioma, and one had a juvenile nasal angiofibroma. In each case, a Tracker No. 18 microcatheter (Target Therapeutics, Inc., Fremont, CA) was navigated into the ophthalmic artery using a steerable guidewire and digital road mapping. Embolic agents included polyvinyl alcohol particles ranging from 350 to 1500 microm in diameter, 2-mm platinum microcoils, and n-butyl-cyanoacrylate. In 12 of 15 cases, lidocaine and amytal provocation tests were conducted before any attempt at embolization to assess the role of the ophthalmic artery in vision. RESULTS: Embolization was successfully performed in the 14 situations in which it was attempted. Positive results of two lidocaine/amytal tests were noted. In one case, embolization was not attempted. In the other case, a larger caliber embolic agent (2-mm platinum coils) was used. A single transient decrease in visual acuity lasting 4 days was the only embolization-related complication. CONCLUSION: Proper case selection, judicious use of embolic agents, and use of provocative testing can result in safe embolization of lesions supplied by the ophthalmic artery. PMID- 9848843 TI - Complex intracranial aneurysms: combined operative and endovascular approaches. AB - OBJECTIVE: Endovascular management of complex intracranial aneurysms is increasingly being considered as an alternative to standard surgical clipping. However, little attention has been paid to the complementary nature of surgery and endovascular therapy. METHODS: Between September 1992 and May 1997, 12 patients with complex intracranial aneurysms were treated with combined operative and endovascular methods. Seven patients demonstrated subarachnoid hemorrhage (two of Grade II, two of Grade III, and three of Grade IV). Five patients demonstrated unruptured aneurysms, i.e., three giant aneurysms (one vertebrobasilar junction aneurysm, one middle cerebral artery bifurcation aneurysm, and one internal carotid artery-ophthalmic artery aneurysm), one large internal carotid artery-ophthalmic artery aneurysm, and one middle cerebral artery serpentine aneurysm. Management strategies involved either surgery followed by endovascular therapy (S-E; n = 5) or endovascular therapy followed by surgery (E-S; n = 7). S-E paradigms included aneurysm exploration followed by endovascular treatment (S-E1; n = 3), partial aneurysm clipping followed by endovascular aneurysm packing (S-E2; n = 1), and extracranial-to-intracranial bypass followed by endovascular parent vessel occlusion (S-E3; n = 1). E-S paradigms included superselective angiography followed by surgical clipping (E S1; n = 2), Guglielmi detachable coil partial dome packing followed by delayed surgical clipping (E-S2; n = 2), proximal temporary vessel balloon occlusion followed by aneurysm clipping (E-S3; n = 2), and proximal permanent vessel occlusion followed by surgical aneurysm decompression for mass effect treatment (E-S4; n = 1). RESULTS: Eleven aneurysms (92%) were completely eliminated. The remaining aneurysm was 90% obliterated and remained quiescent at the 34-month follow-up examination, despite presenting with subarachnoid hemorrhage. No patient experienced repeat bleeding (follow-up period, 23+/-28 mo). There were no deaths. One patient achieved a fair outcome (Glasgow Outcome Scale score of III); all other patients experienced excellent outcomes (Glasgow Outcome Scale score of I). In all cases, the aneurysm management paradigm chosen had a positive effect on definitive therapy. CONCLUSION: Several factors can contribute to the complexity of intracranial aneurysms. Management strategies that combine operative and endovascular techniques in a complementary way, for the best possible outcomes for these patients, can be designed accordingly. PMID- 9848844 TI - Traumatic intracranial carotid tree aneurysms. AB - OBJECTIVE: This study was designed to elucidate the requirements for angiographic evaluation in blunt head injuries, the timing of angiography, and the selection of appropriate therapeutic approaches. METHODS: Twelve cases of traumatic aneurysms (TAs) in the intracranial carotid tree were analyzed in this study. Neurological examination results, computed tomographic scans, pre- and postembolization cerebral angiograms, and follow-up data were included. RESULTS: In 11 of 12 cases, TAs were of cranial base origin; in 1 case, the aneurysm was located in the distal anterior cerebral artery. In seven of the cases with cranial base lesions, aneurysms were located in the intracavernous segment of the internal carotid artery; all of the computed tomographic scans for these cases demonstrated sphenoid sinus wall fractures and hematoma in the sphenoid sinus. In two cases, although the initial angiograms revealed no lesions, a second study performed 2 weeks later demonstrated the presence of aneurysms. Nine of the aneurysms were treated with endovascular techniques, two were managed conservatively, and the remaining one patient died with massive epistaxis while awaiting surgical treatment. No morbidity or additional permanent neurological deficits occurred in the endovascularly treated patient group. CONCLUSION: Patients with head trauma who present with sphenoid sinus fractures and massive epistaxis should be evaluated for the development of TAs as soon as possible. If the patients exhibit fractures without epistaxis, angiography should be deferred for 2 to 3 weeks; if the first angiographic evaluation reveals normal findings, repeated epistaxis should prompt a second angiographic evaluation. Current treatment of TAs involves occlusion of the main artery through the use of endovascular techniques. Cases involving internal carotid artery TAs of cranial base origin and patients who do not tolerate test occlusion require extracranial to-intracranial bypass surgery. PMID- 9848845 TI - The role of endoscopic third ventriculostomy in the management of shunt malfunction. AB - OBJECTIVE: To evaluate the effectiveness of third ventriculostomy as an alternative to shunt revision in the management of shunt malfunction and infection in obstructive hydrocephalus. METHODS: All of the clinical notes of 30 patients treated with third ventriculostomy for malfunctioning or infected shunts between January 1, 1974, and December 31, 1996, were retrospectively reviewed. Third ventriculostomy was performed under fluoroscopic control in the first seven patients and endoscopically in the remainder. A successful outcome was achieved if further shunt revision surgery was avoided. The median follow-up duration was 8.7 years RESULTS: Twenty-three patients (76.7%) experienced successful outcomes, resulting in shunt independence. Of the seven failures, three were technical failures at the time of surgery and the remaining four were manifest within a median of 10 days, resulting in shunt revision. There have been no delayed failures. CONCLUSION: Third ventriculostomy is a valuable alternative to shunt revision in patients affected by obstructive hydrocephalus presenting with shunt malfunction or infection. It should be considered in all suitable cases as the first-line treatment for obstructive hydrocephalus of all causes. Because all failures were manifest within a short time, it is likely that these successes will be durable. PMID- 9848846 TI - Endoscopic neurosurgery and endoscope-assisted microneurosurgery for the treatment of intracranial cysts. AB - OBJECTIVE: Different endoscopic techniques have been introduced into neurosurgery, but accepted terminology and definitions are still missing. We propose a terminology based on whether the endoscope is used alone or in conjunction with an operating microscope and on whether the route of surgical manipulations is through or outside the endoscope. Accordingly, procedures are categorized into endoscopic neurosurgery (EN), endoscope-assisted microneurosurgery (EAM), and endoscope-controlled microneurosurgery (ECM). METHODS: We treated 36 patients with intracranial arachnoid cysts (ACs) and intraventricular cysts endoscopically. The patients ranged in age from 4 months to 69 years (mean age, 31 yr). The follow-up period ranged from 6 to 44 months (mean follow-up duration, 14 mo). The indications were hydrocephalus in 17 patients, focal neurological deficits in 4 patients, progressive nonlocalizing symptomatology in 13 patients, and space occupation in 2 asymptomatic patients. EN was used in 14 cases, EAM in 15 cases, and ECM in 7 cases. RESULTS: The overall success rate was 70%. Nine patients (25%) had unchanged symptomatology, and the condition of two patients (5%) deteriorated. The best success rates were achieved in patients with intraventricular cysts (89%) and posterior fossa ACs (78%). Symptomatic improvement was best achieved in patients with hydrocephalus or focal neurological deficits (81%). CONCLUSION: Different endoscopic techniques (i.e., EN, EAM, and ECM) provide sufficient treatment of selected intracranial cysts. Our data suggest that intraventricular cysts and suprasellar ACs should be approached using EN whereas posterior fossa and sylvian ACs may be more effectively treated using a combined technique (EAM or ECM). PMID- 9848847 TI - Usefulness of heavily T2-weighted magnetic resonance imaging in patients with cerebellopontine angle tumors. AB - OBJECTIVE: Heavily T2-weighted magnetic resonance imaging (MRI) has been reported to be useful for the diagnosis of lesions of the inner ear or its central connections. We evaluated the usefulness of heavily T2-weighted MRI in 18 patients with cerebellopontine angle tumors. METHODS: The lesions were acoustic neuromas in 14 patients and meningiomas in 4 patients. The findings of heavily T2 weighted MRI were compared with those of three-dimensional T1-weighted gradient field echo MRI. RESULTS: An accurate description of the situation of the tumor in the internal auditory canal (IAC) was possible, because the inner ear was clearly shown on the heavily T2-weighted magnetic resonance images. We could therefore detect the acoustic neuromas located at the fundus of the IAC and the meningiomas growing into the IAC. The residual tumor in the IAC can be shown postoperatively. It was difficult to demonstrate these findings using three-dimensional T1 weighted gradient field echo MRI because of the poor presentation of the petrous bone structures. The lower cranial nerves and the VIIth and VIIIth cranial nerves were more clearly demonstrated by heavily T2-weighted MRI than by three dimensional T1-weighted gradient field echo MRI. Using heavily T2-weighted MRI, the VIIth and VIIIth cranial nerves of the lesion side were demonstrated in half of the patients with acoustic neuromas smaller than 2.5 cm. CONCLUSION: We conclude that heavily T2-weighted MRI is useful to detect the relationship among the tumor, the IAC, the inner ear, and the surrounding cranial nerves. PMID- 9848848 TI - Therapeutic strategy for incidentally found pituitary tumors ("pituitary incidentalomas"). AB - OBJECTIVE: To evaluate therapeutic strategy for incidentally found pituitary tumors ("pituitary incidentalomas"), we analyzed the results of magnetic resonance imaging findings and of ophthalmological and endocrinological studies in 28 cases with long-term follow-up (Hardy's classification: Grade A, 24 cases; Grade B, 4 cases). METHODS: Only cases of nonfunctioning macroincidentaloma were analyzed in this study. Cases with ophthalmological and/or endocrinological dysfunction revealed by the first evaluation, even without subjective manifestations, were excluded from the category. Incidentally found functioning tumors were also excluded. RESULTS: The follow-up period ranged from 6 months to 10 years (mean, 5.6 yr). Magnetic resonance imaging and ophthalmological and endocrinological studies, including provocation tests, were conducted once per year. No surgical treatment was required in any case of Grade A tumors and in two cases of Grade B tumors because of no changes revealed by these studies. Transsphenoidal surgery was performed in the remaining two cases of Grade B tumors because of pituitary apoplexy. The second case was one of head injury induced apoplexy. There were no deficits after surgery. The MIB-1 index did not differ in operated incidentaloma and symptomatic pituitary tumors. CONCLUSION: Unless ophthalmological and endocrinological dysfunction is noted, surgical treatment is not required for Grade A pituitary incidentalomas. It is not too late to remove the tumor surgically, even after some dysfunction develops. A patient having a tumor larger than Grade A can still be managed conservatively; however, the patient should be carefully informed of the possibility of pituitary apoplexy, and emergency transsphenoidal surgery is indicated if apoplexy occurs. PMID- 9848849 TI - Combined hyperactive dysfunction syndrome of the cranial nerves: trigeminal neuralgia, hemifacial spasm, and glossopharyngeal neuralgia: 11-year experience and review. AB - OBJECTIVE: A pathological condition caused by vascular compression at the root entry/exit zone of the cranial nerves is designated hyperactive dysfunction syndrome (HDS) of the cranial nerves. Patients with HDS who exhibited a combination of trigeminal neuralgia (TN), hemifacial spasm (HFS), and/or glossopharyngeal neuralgia were retrospectively reviewed, to study the incidence, etiological factors, and demographic characteristics for this combined HDS group. METHODS: Medical and surgical records were analyzed for 41 patients with combined HDS, of 1472 consecutive patients with HDS who were treated between 1984 and 1994. RESULTS: The combined HDS group accounted for 2.8% of all patients with HDS; 19 patients (1.3%) exhibited bilateral symptoms, i.e., 14 cases of TN, 3 of combined TN and HFS, and 2 of HFS. Twenty-two patients (1.5%) exhibited ipsilateral symptoms, i.e., 19 cases of TN and HFS and 3 of TN and glossopharyngeal neuralgia. Excluding three patients whose symptoms were associated with brain tumors or arteriovenous malformations, this patient group was older (63.2 versus 55.3 yr, P = 0.0009) and exhibited an increased percentage of associated hypertension (47.4 versus 17.5%, P = 0.000008), with a female predominance (86.8 versus 71.3%, P = 0.07), compared with the single HDS group. Thirty-six of these patients underwent a total of 61 microvascular decompression procedures, with favorable outcomes. The offending vessels were similar to those in single HDS, which were usually conventional and multiple. CONCLUSION: The associated etiological factors for vascular compression syndromes were more evident in the combined HDS group than in the single HDS group. Progressive arteriosclerotic vasculoarchitectural changes of the vertebrobasilar system, accelerated by aging and hypertension, bring about the development of combined HDS, with a remarkable female predominance. PMID- 9848850 TI - Juxtafacet cysts of the cervical spine. AB - OBJECTIVE: We report a retrospective series of 13 patients who presented with symptoms and signs caused by cervical juxtafacet cysts. Clinical findings, radiographic features, surgical management strategies, and possible causes are reported and discussed. METHODS: We reviewed clinical histories, radiographic studies, surgical notes, and pathological records of all 13 patients who underwent surgery for subaxial cervical juxtafacet cysts from 1984 to 1997 at the Mayo Clinic in Rochester, MN. During the summer of 1997, surgical outcomes were assessed by direct or telephone interview. RESULTS: Nine patients were men and four were women. The average age at the time of presentation was 66 years. One patient had undergone a previous anterior cervical fusion below the level of the cyst. Ten patients presented with radiculopathies. Two patients presented with myeloradiculopathies. One patient presented with a cervical myelopathy. Computed tomographic myelography and magnetic resonance imaging were essential in establishing a preoperative diagnosis. The cysts were located at C7-T1 in nine patients, at C4-C5 in two patients, at C6-C7 in one patient, and at C3-C4 in one patient. All patients underwent posterior laminectomy or hemilaminectomy, excision of the cyst, and decompression of the thecal sac and/or nerve root. Two patients underwent concurrent posterior fusion procedures for instability. All patients experienced good to excellent relief of their radicular pain. All three myelopathies stabilized after surgery. There were no major complications or recurrences. CONCLUSION: Juxtafacet cysts seem to be a degenerative change of the cervical spine rather than a traumatic event. Similar to their counterparts in the lumbar spine, they tend to arise in segments with increased mobility. Surgical treatment is effective. PMID- 9848851 TI - Neurological deterioration as a potential alternative endpoint in human clinical trials of experimental pharmacological agents for treatment of severe traumatic brain injuries. Executive Committee of the International Selfotel Trial. AB - OBJECTIVE: A recently improved understanding of the pathophysiological features of head injuries has led to the development of new drug therapies. Accurate human clinical trials remain necessary to document the efficacy and safety of new agents. It would be helpful to decrease the time from drug development to clinical use and general availability for drugs found to be effective. Conversely, ineffective agents could be abandoned in a timely fashion. RATIONALE: A new endpoint measure, defined as neuroworsening (NW), is an objective observable event that is identifiable during hospitalization. This may enable the efficacy of drugs to be demonstrated or disproved much earlier than with 6-month outcome assessments. The prospective, double-blind, multicenter trial of the N methyl-D-aspartate receptor antagonist Selfotel was used to acquire data on the efficacy of NW in predicting neurological outcomes. The 6-month Glasgow Outcome Scale scores, which were the primary endpoints of that trial, were compared with the frequency of NW. NW was an observable event that could be objectively defined after head injuries. Patients who suffered one or more episodes of NW demonstrated significantly higher morbidity and mortality rates than did patients who did not. CONCLUSION: Future trials should consider the use of NW as an outcome measure that can be included with more traditional measures in the study design. If the strong correlation demonstrated between NW and 6-month Glasgow Outcome Scale scores can be prospectively demonstrated in a successful trial, the time to approval of future agents could be decreased. PMID- 9848852 TI - The biological resolution of malformations of the central nervous system. AB - It has been known for years that the assembly of the nervous system is under genetic control. During the last 10 years, the genes that direct the formation of the brain and spinal cord have begun to be discovered at an amazing pace. Mutations in the fruit fly and advances in molecular genetics have led the way. Gene mutations that cause many of the malformations of the human brain and spinal cord are now known. This has many physician-scientists hoping that an understanding of cause might lead to cure. PMID- 9848853 TI - Reperfusion injury after focal cerebral ischemia: the role of inflammation and the therapeutic horizon. AB - Recent evidence indicates that thrombolysis may be an effective therapy for the treatment of acute ischemic stroke. However, the reperfusion of ischemic brain comes with a price. In clinical trials, patients treated with thrombolytic therapy have shown a 6% rate of intracerebral hemorrhage, which was balanced against a 30% improvement in functional outcome over controls. Destruction of the microvasculature and extension of the infarct area occur after cerebral reperfusion. We have reviewed the existing data indicating that an inflammatory response occurring after the reestablishment of circulation has a causative role in this reperfusion injury. The recruitment of neutrophils to the area of ischemia, the first step to inflammation, involves the coordinated appearance of multiple proteins. Intercellular adhesion molecule-1 and integrins are adhesion molecules that are up-regulated in endothelial cells and leukocytes. Tumor necrosis factor-alpha, interleukin-1, and platelet-activating factor also participate in leukocyte accumulation and subsequent activation. Therapies that interfere with the functions of these factors have shown promise in reducing reperfusion injury and infarct extension in the experimental setting. They may prove to be useful adjuncts to thrombolytic therapy in the treatment of acute ischemic stroke. PMID- 9848854 TI - Analytical evaluation of complex anterior approaches to the cranial base: an anatomic study. AB - OBJECTIVE: The purpose of the present study was to evaluate the anatomic areas of the cranial base exposed by different complex anterior approaches. METHODS: Using 20 embalmed cadaveric heads, we executed five different anterior approaches, i.e., Le Fort 1 approach with splitting or down-fracturing of the hard palate, extended maxillectomy, median mandibulotomy with glossotomy, and mandibular swing transcervical approach. Each approach was performed a minimum of three times. The areas of the intra- and extracranial cranial base exposed by each approach were analytically examined by using a numerical grading system to assess the exposure of major anatomic and neurovascular structures. RESULTS: Good exposure of the midline compartment of the cranial base was provided by the median mandibulotomy with glossotomy and by the Le Fort 1 approach with splitting of the hard palate, whereas the mandibular swing transcervical and extended maxillectomy approaches provided good exposure of the lateral compartment of the cranial base. CONCLUSION: Use of the numerical grading system allowed quantification of the exposure afforded by each approach, with respect to the different compartments (midline and lateral) of the intra- and extracranial cranial base, highlighting the differences among the approaches examined. PMID- 9848855 TI - Systemic administration of the endothelin-A receptor antagonist TBC 11251 attenuates cerebral vasospasm after experimental subarachnoid hemorrhage: dose study and review of endothelin-based therapies in the literature on cerebral vasospasm. AB - OBJECTIVE: Increasing evidence implicates endothelin (ET)-1 in the pathophysiological development of cerebral vasospasm. This study examined the ability of TBC 11251 (TBC), a new ETA receptor antagonist, to prevent vasospasm in a rabbit model of subarachnoid hemorrhage (SAH). METHODS: Eighty-five New Zealand White rabbits were assigned to 1 of 10 groups. SAH was induced by injecting autologous blood into the cisterna magna. The treatment groups were as follows: 1) control (no SAH), 2) SAH alone, 3) SAH plus vehicle every 12 hours (BID), 4) SAH plus 5 mg/kg TBC BID, 5) SAH plus 10 mg/kg TBC BID, 6) SAH plus 20 mg/kg TBC BID, 7) SAH plus vehicle at 24 and 36 hours after SAH (24/36), 8) SAH plus 5 mg/kg TBC 24/36, 9) SAH plus 10 mg/kg TBC 24/36, and 10) SAH plus 20 mg/kg TBC 24/36. Animals were killed 48 hours after SAH, by perfusion-fixation, and then basilar arteries were histologically prepared and their cross-sectional areas were measured. RESULTS: The mean basilar artery cross-sectional area was constricted from 0.332 mm2 in the control group to 0.131 mm2 in the SAH alone group, 0.132 in the vehicle 24/36 group, and 0.125 in the vehicle BID group. All groups treated with TBC showed an increase in cross-sectional luminal basilar artery area, relative to the vehicle-treated groups. The 5 mg/kg TBC BID group exhibited a mean basilar artery area of 0.217 mm2, and the 10 mg/kg TBC BID group showed a mean basilar artery area of 0.240 mm2; both groups were statistically improved, compared with the vehicle-treated groups (P < 0.05). No side effects were seen, and there were no differences in the mean arterial pressures between drug- and vehicle-treated groups. CONCLUSION: These findings demonstrate that systemic administration of the ETA receptor antagonist TBC significantly attenuates cerebral vasospasm after SAH, thus providing additional support for the role of ET-1 in vasospasm. PMID- 9848856 TI - Experimental creation of fusiform carotid artery aneurysms using vein grafts in rats. AB - OBJECTIVE: We developed an in vivo model of growing fusiform aneurysms, using vein grafts to the rat carotid artery. This aneurysm model might demonstrate the pathological features of the development and growth of aneurysms to become giant aneurysms. METHODS: Placement of an interposed femoral vein graft to restore carotid artery flow was performed in Wistar rats. On Day 21, 75% of the grafts (mean diameter, 1.6 mm) were found to be dilated to resemble fusiform aneurysms (mean diameter, 5.82 mm), and 53% of these were giant. Quantitative analysis of the histological findings was performed using image-analyzing software. RESULTS: Histological findings were similar to those for human intracranial giant aneurysms. The average length of the initial grafts in the aneurysm group was 9.1+/-1.9 mm, and grafts were significantly longer and more tortuous than in the normal graft group (6.4+/-0.8 mm) (P = 0.01). Cross-sectional areas of the aneurysms (mean, 18.9 mm2) were significantly correlated with the following: 1) the area of intra-aneurysmal thrombosis (mean, 11.1 mm2) (P < 0.0001); 2) the number of intrathrombotic vascular channels (P = 0.005); and 3) the area of dissection, with hemorrhage, between the thrombus and the wall of the aneurysm (mean, 0.72 mm2) (P = 0.0013). Scanning electron microscopic examination showed evidence of endothelial damage associated with growth of the aneurysms. CONCLUSION: Recurrent hemorrhaging from intrathrombotic vascular channels caused dissection between the thrombus and the aneurysm wall, which led to growth of the experimental aneurysms to giant aneurysms. With this model, we demonstrated the growth mechanism of giant fusiform aneurysms. PMID- 9848857 TI - Inducible nitric oxide synthase expression after traumatic brain injury and neuroprotection with aminoguanidine treatment in rats. AB - OBJECTIVE: We investigated the time course of inducible nitric oxide synthase (iNOS) enzymatic activity and immunocytochemical localization of iNOS expression after traumatic brain injury (TBI), as well as the possible role of iNOS in the pathogenesis of TBI. METHODS: Male Sprague-Dawley rats were anesthetized and underwent moderate parasagittal fluid-percussion brain injury. Rats were decapitated 5 minutes, 6 hours, 1 day, 3 days, 7 days, or 14 days later, and iNOS enzymatic activities were measured (n = 6-8). To determine whether nitric oxide produced by iNOS contributed to the histopathological consequences of TBI, inhibition of iNOS activity using aminoguanidine (intraperitoneal injections of 100 mg/kg aminoguanidine [n = 9] or vehicle [n = 8], twice each day) was conducted for 3 days. RESULTS: Significantly elevated iNOS activity was detected at 3 days (276.8+/-72.3% of contralateral value, means +/- standard errors; P < 0.05), and the most robust increase occurred 7 days after TBI (608.0+/-127.0%, P < 0.01) in the injured parietal cerebral cortex. Immunostaining for iNOS and glial fibrillary acidic protein, at 3 and 7 days after TBI, revealed that the major cellular sources of iNOS expression were cortical Layer 1 astrocytes and macrophages within the subarachnoid space. Administration of aminoguanidine did not reduce contusion volume significantly; however, treatment reduced total cortical necrotic neuron counts (1367.6+/-210.3; P < 0.01, compared with vehicle, 2808.5+/-325.1). CONCLUSION: These data indicate that iNOS is expressed after moderate parasagittal fluid-percussion brain injury, in a time-dependent manner, and that inhibition of iNOS synthesis improves histopathological outcomes. Thus, inhibition of iNOS activation may represent a potential therapeutic strategy for the treatment of TBI. PMID- 9848858 TI - Detection of brain tumor invasion and micrometastasis in vivo by expression of enhanced green fluorescent protein. AB - OBJECTIVE: To determine whether fluorescence from human brain tumor cells transfected with the enhanced green fluorescent protein (EGFP) gene in vitro and xenotransplanted into the brain of nude mice would permit the detection of brain tumor invasion and metastasis in vivo. METHODS: Daoy medulloblastoma cells were transfected with a long terminal repeat-based retroviral vector containing the EGFP gene. Stable EGFP-expressing clones were isolated and stereotactically injected into the frontal cortex of nude mice. Four weeks later, whole brain sections were examined using fluorescence microscopy, immunohistochemistry, and routine hematoxylin and eosin staining for the visualization and detection of tumor cell invasion and metastasis. RESULTS: We demonstrate that EGFP-transduced Daoy cells maintain stable high-level EGFP expression in the central nervous system during their growth in vivo. EGFP fluorescence clearly demarcated the primary tumor margins and readily allowed for the visualization of distant micrometastases and local invasion on the single-cell level. Small metastatic and locally invasive foci, including those immediately adjacent to the tumor's leading invasive edge, were virtually undetectable by routine hematoxylin and eosin staining and immunohistochemistry. EGFP expression also persisted in vitro after cell reculture from brain tissue extracts. CONCLUSION: We show, for the first time, that EGFP-transduced human brain tumor cells can be visualized by fluorescence microscopy after intracerebral implantation. This method is superior to routine hematoxylin and eosin staining and immunohistochemistry for the detection and study of physiologically relevant patterns of brain tumor invasion and metastasis in vivo. PMID- 9848859 TI - The progression of an infundibulum to aneurysm formation and rupture: case report and literature review. AB - OBJECTIVE AND IMPORTANCE: Infundibula (IFs) are funnel-shaped symmetrical enlargements that occur at the origins of cerebral arteries and are apparent on 7 to 25% of otherwise normal angiograms. They are frequently considered as normal anatomic variants of no pathogenic significance. CLINICAL PRESENTATION: We report the case of a ruptured posterior communicating artery aneurysm that had developed at the site of a previously known IF in a 49-year-old hypertensive woman. She had a poor conscious level at admission, with widespread subarachnoid hemorrhage and obstructive hydrocephalus. INTERVENTION: The patient was immediately ventilated, and an external ventricular drain was inserted. The aneurysm was successfully clipped; however, secondary hemorrhage occurred both before and during craniotomy. She developed marked hypernatremia and subsequently died. CONCLUSION: This is the 11th case of IF-to-aneurysm progression reported. It suggests that in certain cases, serial investigations may be indicated with IFs to detect aneurysm formation and preempt rupture. PMID- 9848860 TI - Iatrogenic vertebrobasilar insufficiency after surgery of the subclavian or brachial artery: review of three cases. AB - OBJECTIVE AND IMPORTANCE: Vertebrobasilar insufficiency resulting from disease of the subclavian artery is well recognized. Usually, this occurs as the "subclavian steal" syndrome in the context of chronic subclavian stenosis and is consequently well tolerated because of collateralization. Acute disruption of the hemodynamics of the aortic arch vessels, however, can produce disastrous sequelae. CLINICAL PRESENTATION: We present three cases of iatrogenic vertebrobasilar insufficiency sustained as complications of surgery of the left subclavian artery or its distal continuation. The cases were chosen from a review of approximately 400 emergency neurosurgery consultations requested at the Los Angeles County Hospital between November 1995 and February 1996. INTERVENTION: The first patient underwent repair of a traumatic brachial artery occlusion and awoke postoperatively with bilateral cortical blindness, right hemiparesis, and multiple cranial nerve deficits that were most likely caused by acute subclavian steal. The second underwent removal of a subclavian embolus and developed bilateral cerebellar infarction leading to persistent coma, possibly from inadvertent embolization of the vertebral artery during surgery. The third underwent resection and bypass grafting of a subclavian aneurysm. Good backflow was reported when the vertebral artery was disarticulated from the subclavian artery, and this vessel was not reimplanted into the graft. The patient suffered massive cerebellar infarction leading rapidly to brain death. CONCLUSION: There are myriad ways in which the inherent redundancy of the vertebrobasilar system may be jeopardized, and when this protective mechanism fails, the results can be disastrous. Flow through the vertebral arteries may be compromised by thrombosis, embolization, dissection, inappropriate ligation, excessive head rotation, hypotension, vasospasm, or acute subclavian steal. These examples illustrate the importance of understanding the complex physiology of posterior fossa circulation as the basis of pre-, intra-, and postoperative management of patients undergoing surgery of the subclavian artery. PMID- 9848861 TI - Successful surgical removal of a large arteriovenous malformation in a patient with hemophilia: case report. AB - OBJECTIVE AND IMPORTANCE: This is the first reported case of the successful surgical removal of a large arteriovenous malformation (AVM) in a patient with hemophilia A. CLINICAL PRESENTATION: A 19-year-old male patient was admitted to our department with intracranial hemorrhage. He had previously been diagnosed with hemophilia A and a cerebral AVM. Carotid angiography revealed a large AVM in the right temporal and parietal lobes. The neurological and neuroradiological findings, especially those of single photon emission computed tomography, identified an area of devitalization around the lesion, which was thought to reduce the risk of new deficits resulting from surgical manipulation. INTERVENTION: We resected the AVM in conjunction with supplemental infusions of Factor VIII before, during, and after the operation. A slight cerebral hemorrhage on the 7th postoperative day was observed despite control with Factor VIII, but the patient was discharged without any new deficits. CONCLUSION: We evaluated and managed all problems of a patient with multiple complications and achieved a medical cure. PMID- 9848862 TI - Cerebral fungal infections in the immunocompromised host: a literature review and a new pathogen--Chaetomium atrobrunneum: case report. AB - OBJECTIVE AND IMPORTANCE: A case of a cerebral abscess, occurring in a patient who had undergone bone marrow transplant, caused by a new pathogen, Chaetomium atrobrunneum, and a review of the literature are presented. Although Aspergillus species are by far the most common fungi found in cerebral abscesses in immunocompromised patients, an increasing number of fungi commonly found in nature but not usually associated with infections in humans have been isolated from cerebral abscesses in these patients. CLINICAL PRESENTATION: A 31-year-old male patient, who had undergone a recent bone marrow transplantation for multiple myeloma, presented 3 months after transplantation with right-sided hemiplegia caused by a left parietal hemorrhagic lesion. INTERVENTION: A biopsy guided by computed tomography showed that the abscess contained the fungus C. atrobrunneum. The final identification of the organism was based on cultures, scanning electron microscopic studies, and consultation with the Fungus Testing Laboratory at the University of Texas Health Science Center, San Antonio, TX. The patient was treated with several antifungal medications but developed uncontrolled cerebral edema, which led to further neurological deterioration and eventual death. CONCLUSION: Cerebral abscesses in the immunocompromised patient may no longer be assumed to be caused solely by Aspergillus species. The literature reveals several rare cases of uncommon fungi found in these abscesses. Only four cases of cerebral infections caused by the genus Chaetomium have been reported. The case presented represents the first time a patient who had undergone a bone marrow transplant with a cerebral abscess was found to be caused by the pathogen C. atrobrunneum. PMID- 9848863 TI - Communicating hydrocephalus secondary to diffuse meningeal spread of Wegener's granulomatosis: case report and literature review. AB - OBJECTIVE AND IMPORTANCE: We present a very unusual case of diffuse spread of Wegener's granulomatosis causing hydrocephalus. CLINICAL PRESENTATION: A 53-year old man presented in 1985 with bilateral middle ear infections requiring myringotomies. During the next 18 months, he went on to develop a left Bell's palsy. The patient then began to develop recurrent occipital headaches along with left sixth and seventh nerve palsies and a green nasal discharge requiring hospitalization. Workup included magnetic resonance imaging showing pronounced enhancement of the tentorium and meninges in the occipital region with normal ventricle size. An x-ray of the chest showed multiple pulmonary nodules. A regimen of prednisone and cyclophosphamide was initiated. The patient did well for 2 years until he again developed middle ear infections and headache. Serial lumbar punctures showed increased pressures. A circulating antineutrophil cytoplasmic antibody was positive. Cyclophosphamide was administered, with acetazolamide added for treatment of the elevated intracranial pressure. The patient stabilized for another 2 years but then presented in 1994 with recurrent headache, bilateral papilledema, and mild left arm and right leg weakness. A lumbar puncture was performed with an opening pressure of 52 cm H2O. Computed tomography of the head revealed moderate enlargement of the lateral third and fourth ventricles, consistent with communicating hydrocephalus. INTERVENTION: A right frontal ventriculoperitoneal shunt was placed. A leptomeningeal biopsy performed at the side of catheter placement (far away from any meningeal enhancement revealed by magnetic resonance imaging) showed chronic meningitis and multinucleated giant cells. Cyclophosphamide therapy was begun again. The patient has not experienced recurrence of headache, cranial nerve deficits, or papilledema for more than 3 years. CONCLUSION: This is the first reported case of diffuse involvement of the meninges from Wegener's granulomatosis. Fortunately, this patient responded well to shunting and sustained medical management. Although rare, Wegener's granulomatosis should be included in the differential diagnosis of chronic aseptic meningitis, communicating hydrocephalus, and papilledema. PMID- 9848864 TI - Well-differentiated papillary adenocarcinoma arising in a supratentorial enterogenous cyst: case report. AB - OBJECTIVE AND IMPORTANCE: We report a case of a well-differentiated papillary adenocarcinoma arising in an supratentorial enterogenous cyst. The clinicopathological features of this case and a brief review of the literature are presented. CLINICAL PRESENTATION: A 45-year-old woman presented with abrupt onset of sensory seizures and abnormal sensation on the left side of her face, left leg, and left arm. Radiological studies showed a cystic extraaxial tumor with mass effect in the right parietal area. The initial clinical impression was a metastatic lesion, and a comprehensive metastatic workup revealed no evidence of tumor elsewhere. INTERVENTION: A gross total resection of the solid cystic tumor was achieved by a frontoparietal craniotomy. Sixteen months after the initial surgery, the patient presented with signs of increased intracranial pressure and a large parietal cyst. The cyst was fenestrated at the time of the second craniotomy. CONCLUSION: A pathological study of the initial surgical material revealed it to be a well-differentiated papillary adenocarcinoma in association with an enterogenous cyst. The second surgical specimen consisted only of the benign cyst wall. The patient recovered uneventfully from the second surgery and was free of symptoms 6 months postoperatively. The importance of recognizing the rare possibility of malignant progression of a benign enterogenous cyst in the central nervous system is discussed. PMID- 9848865 TI - Intraventricular perineurioma: case report. AB - OBJECTIVE AND IMPORTANCE: Perineurioma, a rare benign nerve sheath neoplasm occurring in either an intraneural or soft tissue form, has never been reported to arise in the central nervous system. CLINICAL PRESENTATION: We describe the clinical, radiological, and pathological features of a perineurioma arising in the choroid plexus of the third ventricle in a 65-year-old woman and causing obstructive hydrocephalus. INTERVENTION: The lesion, apparently unassociated with a nerve, was gross totally resected by frontal craniotomy using a left-sided transcallosal approach. Short-term follow-up showed no evidence of recurrence. CONCLUSION: Perineurioma of the variety found in soft tissue may occur in the central nervous system, wherein it shows the typical light microscopic, immunohistochemical (epithelial membrane antigen- and Collagen IV-positive, S-100 protein-negative), and ultrastructural (pinocytotic vesicles, discontinuous basement membrane) features. PMID- 9848866 TI - Odontoid process and C1-C2 corrective osteotomy through a posterior approach: technical case report. AB - OBJECTIVE AND IMPORTANCE: To demonstrate a new posterior approach to the anterior elements of the atlas and the axis including the odontoid process. CLINICAL PRESENTATION: A 36-year-old woman presented with ankylosing spondylitis and severe flexion deformity of the cervical spine. She had sustained a trauma 5 years previously, causing the inability to look forward or to open the jaw adequately. An examination demonstrated fixed flexion and rotation of the cervical spine, with no neurological deficit. Radiologically, there was fusion of C1, C2, and the clivus. TECHNIQUE: The upper cervical vertebrae were exposed via a midline posterior incision, the posterior arch of C1 was excised, and the vertebral arteries were mobilized. A wedge osteotomy was performed through the lateral masses of C1 and subsequently through the odontoid. The head was repositioned, and C1-C2 lateral mass screws and a Ransford loop were inserted. CONCLUSION: It is possible to gain sufficient surgical access to the odontoid process via a posterior approach. The technique described is of benefit when the alternative anterior approaches to the upper cervical spine are technically difficult or impossible. PMID- 9848867 TI - The xenon-enhanced computed tomography cerebral blood flow method. PMID- 9848868 TI - Serial magnetic resonance imaging findings for a spontaneously resolving spinal subdural hematoma: case report. PMID- 9848869 TI - Perspectives on neurosurgical practice: neurosurgery in Jordan. PMID- 9848870 TI - Accumulation of p53 and Ki-67 expression do not predict survival in patients with fibrillary astrocytomas or the response of these tumors to radiotherapy. PMID- 9848871 TI - Charles S. Sherrington (1857-1952): historical perspective. PMID- 9848872 TI - Science in neurosurgery: the importance of the scientific method. PMID- 9848873 TI - Inhibition of tumor necrosis factor-alpha- and interleukin-1-induced endothelial E-selectin expression by thiol-modifying agents. AB - The expression of endothelial-leukocyte adhesion molecules has been postulated to be regulated by redox-sensitive events. Tumor necrosis factor-alpha (TNF-alpha)- and interleukin-1 (IL-1)-induced E-selectin expression was analyzed after pretreating human umbilical vein endothelial cells with different thiol-modifying agents, ie, diamide, phenylarsine oxide, N-ethylmaleimide, and diethyl maleate. E selectin protein expression was quantified by indirect immunofluorescence. All compounds suppressed the cytokine-induced E-selectin expression in a concentration-dependent manner, whereas the antioxidant N-acetylcysteine showed no effect. The inhibitory effect of diamide (100 micromol/L, 1 hour) was reversible within 6 hours when the cells were allowed to recover before application of cytokines. Reversibility was strongly delayed when cells were deprived of glutathione by buthionine sulfoximine pretreatment. Glutathione depletion alone did not influence cytokine-induced E-selectin expression. Analysis of cellular glutathione status showed a 3-fold increase in oxidized glutathione after diamide treatment. Monochlorobimane labeling also revealed a decrease in total cellular thiols. During recovery, the glutathione status was restored within 1 hour, whereas total thiol content and E-selectin expression needed at least 6 hours to return to baseline. Complete inhibition of E-selectin expression by the vicinal thiol blocker phenylarsine oxide (0.5 micromol/L) was reversed by dithiols like dithiothreitol or dimercaptopropanol, but not by the monothiol 2-mercaptoethanol. These data suggest that proteins with essential thiols, most probably vicinal thiols. are involved in the IL-1- and TNF-alpha mediated induction of E-selectin. These thiols must be in the reduced state; oxidation or other modification thereof attenuates or abolishes the cells' response to the cytokines. PMID- 9848874 TI - Opposite effects of plasma homocysteine and the methylenetetrahydrofolate reductase C677T mutation on carotid artery geometry in asymptomatic adults. AB - Studies of symptomatic patients have identified hyperhomocysteinemia as an independent risk factor for vascular disease. In case-control studies, a point mutation (C677T) in the gene encoding 5,10-methylenetetrahydrofolate reductase (MTHFR) has also been linked to an increased risk of vascular disease through its effect on homocysteinemia. Our aim was to extend these observations to asymptomatic subjects by studying the influence of both homocysteinemia and its mutation on carotid artery geometry. We examined 144 subjects free of atherosclerotic lesions. Fasting homocysteinemia was measured by high-performance liquid chromatography with fluorometric detection. MTHFR genotype was analyzed by polymerase chain reaction followed by HinfI digestion. Carotid artery geometry was characterized by internal diameter and intima-media thickness, as assessed by a high-resolution echo-tracking system. Subjects in the upper homocysteine tertile had a greater carotid internal diameter than did subjects in the middle and lower tertiles (6516+/-770 versus 6206+/-641 and 5985+/-558 microm, respectively; P<0.001). Subjects homozygous for the mutation had a smaller carotid artery internal diameter than did subjects heterozygous or homozygous for the wild-type allele (5846+/-785 versus 6345+/-673 and 6199+/-671 microm, respectively; P<0.05). Homocysteinemia was not significantly increased in subjects homozygous for the mutation. In multivariate regression analysis, homocysteinemia was independently and positively associated with lumen diameter (P=0.0008) and wall thickness (P=0.020). Conversely, homozygosity for the mutation was negatively associated with internal diameter (P=0.009). These preliminary data suggest that mildly elevated homocysteinemia and homozygosity for the MTHFR C677T mutation are associated with opposite preclinical modifications of carotid artery geometry. If confirmed, these results may have important implications for new treatment strategies for vascular disease before the onset of clinical manifestations. PMID- 9848875 TI - Tibolone prevents atherosclerotic lesion formation in cholesterol-fed, ovariectomized rabbits. AB - Tibolone (Org OD14), a synthetic steroid with estrogenic and progestogenic/androgenic properties, is clinically effective for the treatment of climacteric symptoms and the prevention and treatment of osteoporosis in postmenopausal women. The effect on atherogenesis, however, is not known. In the current study, we investigated the effect of tibolone in comparison with that of estradiol and norethisterone acetate on atherogenesis in 140 ovariectomized New Zealand White rabbits that had been induced by an atherogenic diet (0.4% cholesterol, 20 weeks). Tibolone at 18, 6, or 2 mg/d orally completely prevented cholesterol accumulation and fatty streak formation in the aorta; the impairment of endothelium-dependent smooth muscle relaxation of the aorta; and complex lesion formation after endothelial denudation in the carotid artery. Tibolone also reduced the increased postovariectomy plasma lipid concentrations. Analysis of the results, however, indicated that a substantial part of the strong, beneficial effects were plasma lipid independent. Compared with subcutaneous estradiol decanoate (150 microgram once weekly) and oral 17beta-estradiol (4 mg/d), the effects of tibolone were more pronounced at equipotent uterotropic activity. Norethisterone acetate (1 mg/d) did not affect atherosclerotic lesion formation. There are no indications that the progestogenic/androgenic properties of tibolone counteracted its atheroprotective effect on the vessel wall. Therefore, tibolone has the intrinsic potential to be a compound that protects the arterial vessel wall against atherosclerotic processes. PMID- 9848876 TI - Urokinase receptor-dependent upregulation of smooth muscle cell adhesion to vitronectin by urokinase. AB - The plasminogen activator system has been implicated in the modulation of the response to vascular injury. Although urokinase-type plasminogen activator (uPA) and its receptor (uPAR) may enhance matrix degradation as well as migration and invasion by smooth muscle cells (SMCs), their roles in cell adhesion are uncertain. Therefore, we examined the ability of uPA and uPAR to modulate adhesion of cultured human vascular SMCs to various matrices. We demonstrated a dose-dependent stimulation of adhesion by single-chain uPA (scuPA) to vitronectin (maximum 1.55-fold [+/-0. 04-fold] increase, 10 nmol/L, P<0.002) but not to laminin, collagen I, or collagen IV. Baseline adhesion to vitronectin was completely inhibited by both EDTA and RGD peptide but was restored to >40% of control in the presence of scuPA (P=0.001 and 0.046, respectively). Adhesion to vitronectin was also significantly enhanced by the amino-terminal fragment of uPA (P=0.007) and two-chain, high-molecular-weight uPA (P<0.01) but not by the low molecular-weight fragment of uPA, which lacks the receptor-binding domain. Aprotinin, a plasmin inhibitor, had no effect on baseline or scuPA-stimulated adhesion, suggesting a plasmin-independent process. Preincubation of scuPA with soluble uPAR inhibited scuPA stimulation of adhesion by 88+/-14% (P=0.01), as did pretreatment of SMCs with phosphatidylinositol-specific phospholipase C, which removes glycophosphatidylinositol-anchored proteins, including uPAR. Antibodies to both alphavbeta3 and alphavbeta5 integrin inhibited baseline adhesion but not scuPA stimulation. Finally, coating plates with scuPA alone enabled cell adhesion, which could be inhibited by both soluble uPAR and anti-uPAR antibodies. These data suggest that uPA stimulates adhesion of SMCs specifically to vitronectin and that it is mediated by an interaction with uPAR. Upregulation of both proteins after vascular injury may facilitate migration through stimulation of both matrix degradation and cell adhesion. PMID- 9848877 TI - Expression of thrombomodulin in atherosclerotic lesions and mitogenic activity of recombinant thrombomodulin in vascular smooth muscle cells. AB - Thrombomodulin (TM), a thrombin receptor protein found on the endothelial cell surface, contains 6 tandem epidermal growth factor (EGF)-like structures. Recombinant human TM peptide containing these 6 EGF-like domains (rTME1-6) exhibits mitogenic activity in Swiss 3T3 cells. We examined the localization of TM in atherosclerotic lesions and the effects of rTME1-6 on the growth of cultured rat vascular smooth muscle cells (SMCs). Immunohistochemical analysis demonstrated that TM antigen was localized on monocytes, macrophages, and vascular SMCs. In cultured vascular SMCs, rTME1-6 accelerated [3H]thymidine uptake into DNA in a dose-dependent manner up to 3.4 times the control level. This mitogenic activity was abolished by addition of polyclonal anti-human TM antibody. The rTME1-6-induced mitogenesis was enhanced by EGF. However, a neutralizing monoclonal antibody against the EGF receptor (monoclonal antibody 225) did not inhibit the mitogenic activity of rTME1-6. Calphostin C, a specific protein kinase C inhibitor, and lavendustin-A, an inhibitor of EGF receptor specific protein tyrosine kinase, inhibited the mitogenic activities of both rTME1-6 and EGF. Finally, rTME1-6 treatment increased the level of phosphorylated mitogen-activated protein kinase in SMCs. Together, these results suggest that TM expression in atherosclerotic lesions may be associated with promotion of atherosclerosis through its mitogenic activity in vascular SMCs. PMID- 9848878 TI - Impact of apo(a) length polymorphism and the control of plasma Lp(a) concentrations: evidence for a threshold effect. AB - Plasma lipoprotein(a) [Lp(a)] levels are believed to be controlled predominantly by the apolipoprotein(a) [APO(a)] gene, which encodes the apo(a) glycoprotein, a key constituent of the Lp(a) particle. Previously, it has been accepted that the plasma Lp(a) level is inversely proportional to apo(a) length. To examine this relationship in greater detail, 1500 unrelated, homogeneous (sex, race, age, plasma lipids) subjects were studied, from which 769 were identified with a single-expressing APO(a) allele. A bimodal frequency distribution of apo(a) isoforms was observed. As expected, there was a general inverse relationship between apo(a) isoform size and Lp(a) level. However, when groups with equivalent single-expressing apo(a) isoforms were studied, it was clear that although smaller isoforms were associated on average with higher levels, they were also associated with the greatest variability in level. After logarithmic transformation of Lp(a) data, the overall contribution of the apo(a) length polymorphism was calculated to be 38%. However, in subjects with apo(a) isoforms of 20 K-4 repeats, the corresponding contribution is 10%. We conclude that the contribution of the apo(a) isoform size to the control of plasma Lp(a) level is considerably lower than previously calculated, because the variability in plasma Lp(a) concentration is not uniform across the apo(a) size spectrum. PMID- 9848879 TI - Differences in near-wall shear rate in the carotid artery within subjects are associated with different intima-media thicknesses. AB - In the common carotid artery, reflections originating from the periphery and the flow divider may affect the shape of the flow velocity profile and, hence, near wall shear rate (WSR) differently just before the bifurcation (location B) than 20 to 30 mm farther upstream (location A). Recent developments in ultrasound technology allow the assessment of WSR and intima-media thickness (IMT) at the same site in the carotid artery in vivo. We therefore determined WSR at locations A and B and investigated whether the differences between both sites, if any, were associated with different IMTs and different mechanical properties of the arterial wall. The effect of age on the possible differences was assessed as well. The study was performed on presumably healthy volunteers (n=53). In all individuals, IMT was larger at location B than at location A. The relative difference in IMT between both locations was not affected by age. No significant differences in diameter and distension were found between locations. Near peak systolic and near mean WSR at the posterior wall (PWSRp and MWSRp, respectively) were significantly lower at location B than at location A. The relative differences in PWSRp and MWSRp between both locations within subjects were independent of age. The velocity profiles were more blunted at location A than at location B. PWSRp and MWSRp significantly decreased and IMT significantly increased with age at both locations. IMT was negatively correlated with PWSRP and MWSRP at location B, but this correlation was not significant at location A. In summary, in the common carotid artery, the lower WSR near the bifurcation, as compared with 20 to 30 mm upstream, is associated with a larger IMT than at the more proximal site. The relative difference between both locations within subjects is independent of age. PMID- 9848880 TI - Arterial injury by cholesterol oxidation products causes endothelial dysfunction and arterial wall cholesterol accumulation. AB - Cholesterol oxidation products (ChOx) have been reported to cause acute vascular injury in vivo; however, the pharmacokinetics of ChOx after administration and the mechanisms by which they cause chronic vascular injury are not well understood. To further study the pharmacokinetics and atherogenic properties of ChOx, New Zealand White rabbits were injected intravenously (70 mg per injection, 20 injections per animal) with a ChOx mixture having a composition similar to that found in vivo during a 70-day period. Total ChOx concentrations in plasma peaked almost immediately after a single injection, declined rapidly, and returned to preinjection levels in 2 hours. After multiple injections, the ChOx concentrations rose gradually to levels 2- to 3-fold above baseline levels, increasing mostly in the cholesteryl ester fraction of LDL and VLDL. Rabbit serum and the isolated LDL/VLDL fraction containing elevated ChOx concentrations were cytotoxic to V79 fibroblasts and rabbit aortic endothelial cells. At the time of killing, cholesterol levels in the aortas from ChOx-injected rabbits were significantly elevated despite the fact that plasma cholesterol levels remained in the normal range. In addition, aortas from the ChOx-injected rabbits retained more 125I-labeled horseradish peroxidase, measured 20 minutes after intravenous injection. Transmural concentration profiles across the arterial wall also showed increased horseradish peroxidase accumulation in the inner half of the media from the thoracic aorta in ChOx-injected rabbits. In conclusion, ChOx injection resulted in accumulation of circulating ChOx and induced increased vascular permeability and accumulation of lipids and macromolecules. This study reveals that even under normocholesterolemic conditions, ChOx can cause endothelial dysfunction, increased macromolecular permeability, and increased cholesterol accumulation, parameters believed to be involved in the development of early atherosclerotic lesions. PMID- 9848881 TI - Serum homocysteine and risk of coronary heart disease and cerebrovascular disease in elderly men: a 10-year follow-up. AB - Hyperhomocysteinemia is an independent risk factor for atherosclerotic disease in the middle-aged. We investigated whether a high serum homocysteine level is a risk factor for vascular disease in 878 elderly men (mean age at baseline, 71.5 years; range, 64 to 84 years) in a population-based, representative cohort followed up for 10 years in Zutphen, the Netherlands. Thirty-one percent had nonfasting homocysteine levels >/=17 micromol/L. After adjustment for other major risk factors, high homocysteine levels at baseline (the third compared with the first tertile) were associated with an increased baseline prevalence of myocardial infarction (odds ratio [OR], 1.81; 95% confidence interval [CI], 1.07 to 3.08; P for trend, 0.03) and with a marginally significant increase in the risk of dying of coronary heart disease (relative risk [RR], 1.58; 95% CI, 0.93 to 2.69; P for trend, 0.09) but not with an increased risk of first-ever myocardial infarction. In addition, high homocysteine levels at baseline were associated with an increased baseline prevalence of stroke (OR, 4.61; 95% CI, 1.79 to 11.89; P for trend, 0.002) and with an increased risk of dying of cerebrovascular disease in subjects without hypertension (RR, 6.18; 95% CI, 2.28 to 16.76) but not in those with hypertension. High homocysteine levels were associated with an increased risk of first-ever stroke among normotensive subjects that was not statistically significant (RR, 1. 77 [95% CI, 0.83 to 3.75; P for trend, 0.14]). In a general population of elderly men, a high homocysteine level is common and is strongly associated with the prevalence of coronary heart disease and cerebrovascular disease. It is a strong predictive factor for fatal cerebrovascular disease in men without hypertension but less so for coronary heart disease. PMID- 9848882 TI - Hormone replacement therapy improves cardiovascular risk by lowering plasma viscosity in postmenopausal women. AB - Hormone replacement therapy may protect against cardiovascular disease through several mechanisms that have variable actions on the major determinants of plasma viscosity. Plasma viscosity is an important predictor of incident and recurrent cardiovascular events and mortality in coronary heart disease patients. The effect of estrogen alone or in combination with progestin on plasma viscosity is not known. Using a randomized, double-blind design, we examined the impact of the following daily hormone regimens on plasma viscosity in 23 women: (1) 1 mg estradiol and 2.5 mg medroxyprogesterone (n=7); (2) 1 mg estradiol alone (n=8); and (3) placebo (n=8). Plasma viscosity, fibrinogen, and standard lipoprotein levels were determined at baseline and after 12 weeks of intervention. Plasma viscosity was measured at 37 degreesC with a coaxial microviscometer. Fibrinogen was measured by the Clauss method. Significant changes in plasma viscosity (mPa.s) levels occurred among treatment groups (P<0.01) after the intervention. Plasma viscosity was significantly reduced with estrogen replacement therapy (P<0.01). These data demonstrate that estrogen replacement therapy lowers plasma viscosity. This study suggests an additional mechanism for the cardiovascular protection conferred to postmenopausal women on estrogen replacement therapy. PMID- 9848883 TI - The HMG-CoA reductase inhibitor atorvastatin increases the fractional clearance rate of postprandial triglyceride-rich lipoproteins in miniature pigs. AB - We have previously shown in vivo that the 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor atorvastatin decreases hepatic apolipoprotein B (apoB) secretion into plasma. To test the hypothesis that atorvastatin modulates exogenous triglyceride-rich lipoprotein (TRL) metabolism in vivo, an oral fat load (2 g fat/kg body wt) containing retinol (50 000 IU) was given to 6 control miniature pigs and to 6 animals after 28 days of treatment with atorvastatin 3 mg. kg-1. d-1. A multicompartmental model was developed by use of SAAM II and kinetic analysis performed on the plasma retinyl palmitate (RP) data. Peak TRL (d<1.006 g/mL; Sf>20) triglyceride concentrations were decreased 29% by atorvastatin, and the time to achieve this peak was delayed (5.2 versus 2.3 hours; P<0.01). The TRL triglyceride 0- to 12-hour area under the curve was decreased by 24%. In contrast, atorvastatin treatment had no effect on peak TRL RP concentrations, time to peak, or its rate of appearance into plasma; however, the TRL RP 0- to 12-hour area under the curve was decreased by 20%. Analysis of the RP kinetic parameters revealed that the TRL fractional clearance rate was increased significantly, 1.4-fold (3.093 versus 2.276 pools/h; P=0.012), with atorvastatin treatment. The percent conversion of TRL RP from the rapid-turnover to the slow-turnover compartment was decreased by 47% with atorvastatin treatment. The TRL RP fractional clearance rate was negatively correlated with very low density lipoprotein apoB production rate measured in the fasting state (r=-0.49). Thus, although atorvastatin had no effect on intestinal TRL assembly and secretion, plasma TRL clearance was significantly increased, an effect that may relate to a decreased competition for removal processes by hepatic very low density lipoprotein. PMID- 9848884 TI - Plasma extracellular superoxide dismutase levels in an Australian population with coronary artery disease. AB - In vitro experiments suggest that free radicals may contribute importantly to atherogenesis. Superoxide dismutase (SOD), particularly extracellular SOD (EC SOD), which accounts for the majority of SOD biological activity, is a major superoxide scavenger. We explored factors that may affect plasma EC-SOD levels measured by ELISA and assessed the association between plasma EC-SOD and coronary artery disease documented angiographically in 590 white Australian patients 400 ng/mL were heterozygous for the Arg213-->Gly mutation at the EC-SOD gene; there was also a positive correlation with age (r=0.131, P=0.0016). Plasma EC-SOD in current smokers (75. 0+/-9.3 ng/mL) was much lower than in nonsmokers (111.7+/-8.2 ng/mL, P<0.01), and ex-smokers had intermediate levels (84.3+/-7.1 ng/mL). Levels were significantly lower in patients with than in those without a history of acute myocardial infarction (MI) (76.1+/-7.5 versus 110.1+/-6.0 ng/mL, P<0.05), and low plasma EC-SOD was independently associated with an increased likelihood of a history of MI (OR, 2.04; 95% CI, 1.10 to 3.82); higher EC-SOD levels also tended to be associated with delayed onset of MI. In conclusion, our study establishes that in patients assessed by coronary angiography, circulating EC-SOD is lower in men than in women and in smokers of each sex and that low levels are independently associated with a history of MI. These findings are consistent with EC-SOD's being protective and contributing to reduced coronary risk. PMID- 9848885 TI - Upregulation of prostacyclin synthesis-related gene expression by shear stress in vascular endothelial cells. AB - Prostacyclin (prostaglandin I2, PGI2) has a variety of functions, including inhibition of smooth muscle cell proliferation, vasodilation, and antiplatelet aggregation. PGI2 production in endothelial cells has been reported to increase biphasically after shear loading, but the underlying mechanism is not well understood. To clarify the mechanism for the second phase of PGI2 upregulation, we examined the gene expression of the enzymes involved in PGI2 production in human umbilical vein endothelial cells (HUVECs) after shear-stress (24 dyne/cm2) loading. The production of 6-keto-PGF1alpha, a stable metabolite of PGI2, increased time-dependently under shear stress. The arachidonic acid liberation from membrane phospholipids in HUVECs after 12 hours of shear loading was increased significantly compared with the static condition. No change was observed for cytosolic phospholipase A2 expression, as detected by reverse transcription-polymerase chain reaction and Western blotting. Cyclooxygenase (COX)-1 mRNA increased after 1 hour of shear loading, and the increase lasted for 12 hours, the longest time tested, whereas COX-2 mRNA increased after 1 hour of shear loading and peaked at 6 hours. An increase of COX-1 expression was detected at 12 hours of shear loading by Western blotting. No expression of COX-2 was detected in the static control, but induced expression was observed at 6 hours after shear loading. PGI2 synthase was also found to be upregulated. These results suggest that the elevated PGI2 production by shear stress is mediated by increased arachidonic acid release and a combination of increased expression of COXs and PGI2 synthase. PMID- 9848886 TI - Direct assessment of lipoprotein outflow from in vivo-labeled arterial tissue as determined in an in vitro perfusion system. AB - The rate of cholesterol deposition during the atherosclerotic process is determined by the balance between the inflow and outflow of plasma lipoproteins in the arterial wall. Whereas the rate of inflow may be measured directly, the rate of outflow has most often been calculated indirectly from lipoprotein uptake by using the 2-compartment model. One objection against such calculations is that lipoprotein binding is not being considered. In the present study 2 different protocols were used to obtain a direct measure of the outflow of lipoproteins from atherosclerotic rabbit aortas. Thus, 3 rabbits with experimental atherosclerosis were given 125I-LDL intravenously and 3 were given [14C]cholesterol perorally. Twenty-four hours later the aortas were removed and the outflow of label was monitored during in vitro perfusion. Despite the different protocols, our results were consistent and indicated that fractional loss relative to whole tissue was approximately 0.01 pool/h, which is 1 order of magnitude lower than current estimates based on the 2-compartment model (0.04 to 0.4 pool/h). Furthermore, whereas as much as 2/3 to 3/4 of the tracer that had entered the arterial wall was effectively trapped, the remainder equilibrated at a faster rate (0.06 pool/h). In conclusion, it seems that tissue binding constitutes a prominent and possibly underrated mechanism of lipoprotein deposition, at least in the atherosclerotic rabbit aorta. Furthermore, this means that current estimates of lipoprotein exchange parameters based on the 2 compartment model (eg, fractional loss) may rest on invalid assumptions and should be regarded with caution. PMID- 9848887 TI - Phospholipase A2 type II binds to extracellular matrix biglycan: modulation of its activity on LDL by colocalization in glycosaminoglycan matrixes. AB - We recently reported the presence of secretory, nonpancreatic phospholipase A2 type II (snpPLA2; EC 3.1.1.4) in human atherosclerotic arteries (Hurt-Camejo et al, Arterioscler Thromb Vasc Biol. 1997;17:300-309). SnpPLA2 may generate the proinflammatory products lysophospholipids and free fatty acids, thus contributing to atherogenesis when acting on low density lipoproteins (LDLs) retained in the arterial wall. Immunohistochemical studies showed that smooth muscle cells (SMCs) in human arterial tissue are the main sources of snpPLA2. In cultures of human arterial SMCs, snpPLA2 interacts with versican and smaller heparan/chondroitin sulfate proteoglycans (PGs) secreted as soluble components into the medium. In the present study, we investigated the binding of snpPLA2 to extracellular matrix (ECM) PGs produced by SMCs. The results show that snpPLA2 can bind to the ECM at physiological salt concentrations. ECM-bound snpPLA2 was active, hydrolyzing phosphatidylcholine-containing micelles. Soluble chondroitin 6-sulfate at concentrations >1 micromol/L, but not heparin or heparan sulfate, was able to release ECM-bound snpPLA2. The PG mainly involved in the binding of snpPLA2 was identified as biglycan. Perlecan was also present in the ECM synthesized by SMCs, but it contributed less to the binding of snpPLA2. Experiments with immobilized glycosaminoglycans indicated that snpPLA2 hydrolyzed 7-fold more LDL phospholipids when the lipoprotein and the enzyme were colocalized in a matrix with chondroitin-6-sulfate compared with one with heparin. These data suggest that retention of snpPLA2 in ECMs of different composition may modulate the enzymatic activity of snpPLA2 toward LDL. The results presented in this work support the hypothesis of the potential contribution of snpPLA2 to atherosclerosis. PMID- 9848888 TI - Effect of the novel antiplatelet agent cilostazol on plasma lipoproteins in patients with intermittent claudication. AB - Cilostazol is an antiplatelet agent and vasodilator marketed in Japan for treatment of ischemic symptoms of peripheral vascular disease. It is currently being evaluated in the United States for treatment of symptomatic intermittent claudication (IC). Cilostazol has been shown to improve walking distance in patients with IC. In addition to its reported vasodilator and antiplatelet effects, cilostazol has been proposed to have beneficial effects on plasma lipoproteins. We examined the effect of cilostazol versus placebo on plasma lipoproteins in 189 patients with IC. After 12 weeks of therapy with 100 mg cilostazol BID, plasma triglycerides decreased 15% (P<0.001). Cilostazol also increased plasma high density lipoprotein cholesterol (HDL-C) (10%) and apolipoprotein (apo) A1 (5.7%) significantly (P<0.001 and P<0.01, respectively). Both HDL3 and HDL2 subfractions were increased by cilostazol; however, the greatest percentage increase was observed in HDL2. Individuals with baseline hypertriglyceridemia (>140 mg/dL) experienced the greatest changes in both HDL-C and triglycerides with cilostazol treatment. In that subset of patients, HDL-C was increased 12.2% and triglycerides were decreased 23%. With cilostazol, there was a trend (3%) toward decreased apoB as well as increased apoA1, resulting in a significant (9.8%, P<0.002) increase in the apoA1 to apoB ratio. Low density lipoprotein cholesterol and lipoprotein(a) concentrations were unaffected. Cilostazol treatment resulted in a 35% increase in treadmill walking time (P=0.0015) and a 9.03% increase in ankle-brachial index (P<0.001). These results indicate that in addition to improving the symptoms of IC, cilostazol also favorably modifies plasma lipoproteins in patients with peripheral arterial disease. The mechanism of this effect is currently unknown. PMID- 9848889 TI - Adsorption of fibrinogen to droplets of liquid hydrophobic phases. Functionality of the bound protein and biological implications. AB - Fibrinogen adsorbs spontaneously from aqueous media containing that protein to droplets of liquid hydrophobic phases dispersed in those same media. Examples of such phases include mineral oils, straight-chain hydrocarbons, and various plant- and animal-derived oils. Lecithin preexisting on the surface of oil droplets reduces significantly the amount of fibrinogen that can otherwise bind to them. When bound, fibrinogen remains active in the classic sense of fibrin gelation. As a consequence, oil droplets coated with fibrinogen can participate in a host of biologically important adhesive processes in which the protein would be expected to participate. Certain polyanions, eg, heparin, pentosan polysulfate, dextran sulfate, and suramin, bind to adsorbed fibrin(ogen) and prevent thrombin dependent adhesion of fibrinogen-coated surfaces. Thus, these polyanions can be used to prevent adhesion between fibrin(ogen)-coated oil droplets and other fibrin(ogen)-coated surfaces. Potential practical applications and biological implications of these phenomena are presented and discussed. PMID- 9848890 TI - Carotid and femoral artery wall thickness and stiffness in patients at risk for cardiovascular disease, with special emphasis on hyperhomocysteinemia. AB - Recent developments in ultrasound technology enable the noninvasive measurement of structural and functional vessel wall changes. Until now, the effect of homocysteine on the arterial wall has remained unclear: reports on intima-media thickness (IMT) yield conflicting results, whereas data on vessel wall stiffness are lacking. Because several cardiovascular risk factors result in an increased IMT or stiffness, different groups at risk for atherosclerotic disease, with special emphasis on hyperhomocysteinemia, were studied. Nineteen patients homozygous and 14 subjects heterozygous for cystathionine beta-synthase (CBS) deficiency, 21 patients with familial hypercholesterolemia (FH), 15 patients with essential hypertension, 20 smokers, and 28 control subjects were studied. The IMT values (both right and left) of the common carotid artery (CCA), bulb (BUL), internal carotid artery (ICA), and common femoral artery (CFA) were measured in millimeters by high-resolution ultrasound (Biosound). The distensibility (DC, in 10(-3). kPa-1) and compliance (CC in mm2. kPa-1) coefficients of the CCA (right and left) and CFA (right) were determined by a wall track system (Pie Medical). The mean IMT of the posterior wall in the CCA was 0.70+/-0.09 mm in healthy controls. For patients with vascular disease, FH, and hypertension and in smokers, the mean CCA IMT was larger, whereas no major differences in IMT were observed in patients either homozygous or heterozygous for CBS deficiency. The DC and CC in the right CCA were 23.5+/-6.9 (10(-3). kPa-1) and 0.9+/-0.3 (mm2. kPa 1) in healthy subjects, slightly lower in patients homozygous for CBS deficiency, and clearly lower in patients with vascular disease, FH, and hypertension. No positive correlation was found between plasma homocysteine level and either IMT, CC, or DC. Because smoking was a confounder in each risk group, a stepwise regression analysis was carried out to assess the contribution of each risk factor on IMT and arterial wall stiffness. Age explained most of the variation in IMT of the CCA (coefficient of determination R2 of 0.34), whereas R2 values for serum low density lipoprotein cholesterol, smoking (pack-years), and systolic blood pressure were 0.08, 0.07, and 0.06, respectively. Homocysteine did not contribute to variation in IMT in both the CCA and CFA. Age and smoking contributed to the variation in IMT in the CFA. The variation in DC and CC in the right CCA and right CFA could in part be explained by age, low density lipoprotein cholesterol, and blood pressure. Plasma homocysteine concentration explained only a small proportion of the variation in DC in the CCA (R2=0.02) and in CC in the CFA (R2=0.04). In this study, no relationship was found between homocysteine level and the thickness of the arterial wall, with only a marginal influence on stiffness. PMID- 9848891 TI - Fibronectin and collagen I matrixes promote calcification of vascular cells in vitro, whereas collagen IV matrix is inhibitory. AB - Vascular calcification is a frequent component of atherosclerosis, yet the pathological mechanisms that regulate its formation are poorly understood. Calcification of the vessel wall may represent a process by which cells that normally exhibit a smooth muscle phenotype differentiate into cells that exhibit an osteoblast-like phenotype. One of the determinants of cellular phenotype is extracellular matrix; thus, we undertook the current study to evaluate the influence of extracellular matrix on calcification of vascular cells in vitro. Cell lines derived from bovine aortic media were divided into 1 of 3 groups: those that did not mineralize, those that mineralized slowly, or those that mineralized rapidly. When slowly mineralizing cells were plated onto matrix produced by rapidly mineralizing cells, the time required for mineralization decreased from 33+/-3.0 days to 7.8+/-1.3 days. Matrix produced by rapidly mineralizing cells was found to contain 3 times the amount of collagen I and fibronectin but 70% less collagen IV than nonmineralizing clones. When slowly mineralizing cells were cultured on purified collagen I or fibronectin, mineralized nodule formation, calcium incorporation, von Kossa staining, and alkaline phosphatase activity increased. In contrast, culturing slowly mineralizing cells on purified collagen IV inhibited these mineralization parameters. Furthermore, blocking antibodies to alpha5 integrins significantly inhibited the fibronectin-mediated increases in alkaline phosphatase activity, indicating that integrin-based signaling may be involved. These data suggest that matrix composition can regulate development of arterial calcification and that a subpopulation of vascular cells preferentially produces positively regulating matrix components. PMID- 9848892 TI - Immunization of LDL receptor-deficient mice with homologous malondialdehyde modified and native LDL reduces progression of atherosclerosis by mechanisms other than induction of high titers of antibodies to oxidative neoepitopes. AB - We and others previously showed that immunization of rabbits with different forms of oxidized low density lipoprotein (LDL) significantly reduced atherogenesis. We now investigated the effect of continued immunization on atherosclerosis in LDL receptor-deficient (LDLR-/-) mice to determine whether a similar reduction of atherosclerosis occurred in murine models and whether this was due to humoral immune responses, ie, formation of high titers of antibodies to oxidation specific epitopes. Three groups of LDLR-/- mice were repeatedly immunized with homologous malondialdehyde-modified LDL (MDA-LDL), native LDL, or phosphate buffered saline (PBS) for 7 weeks. Extensive hypercholesterolemia and accelerated atherogenesis were then induced by feeding a cholesterol-rich diet for 17 weeks, during which immunizations were continued. Binding of immunoglobulin (Ig) M and IgG antibodies, as well as IgG1 and IgG2a isotypes, to several epitopes of oxidized LDL were followed throughout the study. After 24 weeks of intervention, atherosclerosis in the aortic origin was significantly reduced by 46.3% and 36.9% in mice immunized with MDA-LDL and native LDL, respectively, compared with PBS (133 558 and 157 141 versus 248 867 microm2 per section, respectively). However, the humoral immune response to oxidative neoepitopes in the MDA-LDL group was very different from that of the LDL or PBS group. IgG antibody binding to MDA-LDL and other epitopes of oxidized LDL, such as oxidized phospholipid (cardiolipin), oxidized cholesterol, or oxidized cholesteryl linoleate, but not native LDL, increased markedly in mice immunized with MDA-LDL, but not in mice immunized with native LDL or PBS. In the MDA-LDL group, both T helper cell (Th)2-dependent IgG1 antibody and Th1-dependent IgG2a antibody binding to oxidative neoepitopes increased significantly over time. The fact that mice immunized with both MDA-LDL and native LDL had a significant reduction in atherosclerosis, whereas only the MDA-LDL group developed very high titers of antibodies to oxidation-specific epitopes, suggests that the antiatherogenic effect of immunization is not primarily dependent on very high titers of antibodies to oxidation-specific epitopes but is more likely to result from the activation of cellular immune responses. PMID- 9848893 TI - Chemokine receptor CCR2 expression and monocyte chemoattractant protein-1 mediated chemotaxis in human monocytes. A regulatory role for plasma LDL. AB - The subendothelial accumulation of macrophage-derived foam cells is one of the hallmarks of atherosclerosis. The recruitment of monocytes to the intima requires the interaction of locally produced chemokines with specific cell surface receptors, including the receptor (CCR2) for monocyte chemoattractant protein-1 (MCP-1). We have previously reported that monocyte CCR2 gene expression and function are effectively downregulated by proinflammatory cytokines. In this study we identified low density lipoprotein (LDL) as a positive regulator of CCR2 expression. Monocyte CCR2 expression was dramatically increased in hypercholesterolemic patients compared with normocholesterolemic controls. Similarly, incubation of human THP-1 monocytes with LDL induced a rapid increase in CCR2 mRNA and protein. By 24 hours the number of cell surface receptors was doubled, causing a 3-fold increase in the chemotactic response to MCP-1. The increase in CCR2 expression and chemotaxis was promoted by native LDL but not by oxidized LDL. Oxidized LDL rapidly downregulated CCR2 expression, whereas reductively methylated LDL, which does not bind to the LDL receptor, had only modest effects on CCR2 expression. A neutralizing anti-LDL receptor antibody prevented the effect of LDL, suggesting that binding and internalization of LDL were essential for CCR2 upregulation. The induction of CCR2 expression appeared to be mediated by LDL-derived cholesterol, because cells treated with free cholesterol also showed increased CCR2 expression. These data suggest that elevated plasma LDL levels in conditions such as hypercholesterolemia enhance monocyte CCR2 expression and chemotactic response and potentially contribute to increased monocyte recruitment to the vessel wall in chronic inflammation and atherogenesis. PMID- 9848894 TI - Antenatal screening for syphilis. Still important in preventing disease. PMID- 9848895 TI - Proton pump inhibitors may mask early gastric cancer. Dyspeptic patients over 45 should undergo endoscopy before these drugs are started. PMID- 9848896 TI - Uncoupling proteins: the unravelling of obesity? Increased understanding of mechanisms may lead, in time, to better drugs. PMID- 9848897 TI - Emotional wellbeing and its relation to health. Physical disease may well result from emotional distress. PMID- 9848898 TI - Paracetamol (acetaminophen) poisoning. No need to change current guidelines to accident departments. PMID- 9848899 TI - Syphilis in pregnant women and their children in the United Kingdom: results from national clinician reporting surveys 1994-7. AB - OBJECTIVE: To measure the incidence of syphilis detected in pregnancy and congenital syphilis in the United Kingdom. DESIGN: Surveys through consultants in genitourinary medicine and paediatricians with active surveillance. SETTING: United Kingdom, 1994-7. SUBJECTS: Women treated for syphilis in pregnancy, and children with early congenital syphilis born in the United Kingdom. RESULTS: Over 3 years 139 women were diagnosed with and treated for syphilis in pregnancy; 121 were detected through antenatal screening. Thirty one had confirmed or probable congenitally transmissible syphilis, putting their pregnancies at risk. These were minimum figures but are compatible with the 90 to 100 women newly diagnosed annually as having infectious or early latent syphilis. A universal screening policy would require 18 600 and 55 700 women (maximum numbers) to be screened, respectively, to detect one woman needing treatment and to prevent one case of congenital syphilis. Nine presumptive cases of children with congenital syphilis born in the United Kingdom were reported. Mothers requiring treatment for syphilis were found in almost every health region but were more prevalent in London and the south east. Being born abroad and belonging to an ethnic minority group were strong risk factors, but 14% (19 of 121) of cases treated and six of 31 definite or probably transmissible cases occurred in white women born in the United Kingdom. CONCLUSIONS: Congenitally transmissible syphilis continues to occur among pregnant women in the United Kingdom. Cases would be missed and stillbirths and congenitally infected babies would occur if antenatal screening was abandoned. PMID- 9848900 TI - Prospective study of post-traumatic stress disorder in children involved in road traffic accidents. AB - OBJECTIVE: To determine the prevalence of severe psychological trauma --that is, post-traumatic stress disorder--in children involved in everyday road traffic accidents. DESIGN: 12 month prospective study. SETTING: Accident and emergency department, Royal United Hospital, Bath. SUBJECTS: 119 children aged 5-18 years involved in road traffic accidents and 66 children who sustained sports injuries. MAIN OUTCOME MEASURE: Presence of appreciable psychological distress; fulfillment of diagnostic criteria for post-traumatic stress disorder. RESULTS: Post traumatic stress disorder was found in 41 (34.5%) children involved in road traffic accidents but only two (3.0%) who sustained sports injuries. The presence of post-traumatic stress disorder was not related to the type of accident, age of the child, or the nature of injuries but was significantly associated with sex, previous experience of trauma, and subjective appraisal of threat to life. None of the children had received any psychological help at the time of assessment. CONCLUSIONS: One in three children involved in road traffic accidents was found to suffer from post-traumatic stress disorder when they were assessed 6 weeks after their accident. The psychological needs of such children after such accidents remain largely unrecognised. PMID- 9848902 TI - Community based study of treatment seeking among subjects with symptoms of sexually transmitted disease in rural Uganda. PMID- 9848901 TI - Intranasal corticosteroids versus oral H1 receptor antagonists in allergic rhinitis: systematic review of randomised controlled trials. AB - OBJECTIVE: To determine whether intranasal corticosteroids are superior to oral H1 receptor antagonists (antihistamines) in the treatment of allergic rhinitis. DESIGN: Meta-analysis of randomised controlled trials comparing intranasal corticosteroids with oral antihistamines. SETTING: Randomised controlled trials conducted worldwide and published between 1966 and 1997. SUBJECTS: 2267 subjects with allergic rhinitis in 16 randomised controlled trials. MAIN OUTCOME MEASURES: Nasal blockage, nasal discharge, sneezing, nasal itch, postnasal drip, nasal discomfort, total nasal symptoms, nasal resistance, and eye symptoms and global ratings. Outcomes measured on different scales were combined to determine pooled odds ratios (categorical outcomes) or standardised mean differences (continuous outcomes). Assessment of heterogeneity between studies, and subgroup analyses of eye symptoms, were undertaken. RESULTS: Intranasal corticosteroids produced significantly greater relief than oral antihistamines of nasal blockage (standardised mean difference 0.63, 95% confidence interval - 0.73 to - 0.53), nasal discharge (-0.5, - 0.6 to - 0.4), sneezing (- 0.49, - 0.59 to - 0.39), nasal itch (- 0.38,- 0.49 to - 0.21), postnasal drip (- 0.24,- 0.42 to - 0.06), and total nasal symptoms (- 0.42,- 0.53 to - 0.32), and global ratings gave an odds ratio for deterioration of symptoms of 0.26 (0.08 to 0.8). There were no significant differences between treatments for nasal discomfort, nasal resistance, or eye symptoms. The effects on sneezing, total nasal symptoms, and eye symptoms were significantly heterogeneous between studies. Other combined outcomes were homogeneous between studies. Subgroup analysis of the outcome of eye symptoms suggested that the duration of assessment (averaged mean score over the study period versus mean score at end of study period) might have accounted for the heterogeneity. CONCLUSION: The results of this systematic review, together with data on safety and cost effectiveness, support the use of intranasal corticosteroids over oral antihistamines as first line treatment for allergic rhinitis. PMID- 9848903 TI - Prolonged urticaria with 17-1A antibody. PMID- 9848904 TI - The new NHS: moscow calling PMID- 9848906 TI - Not born to be idle PMID- 9848905 TI - Genetic discrimination in life insurance: empirical evidence from a cross sectional survey of genetic support groups in the United Kingdom. AB - OBJECTIVES: To gather empirical evidence on any discrimination based on genetic information shown by the insurance industry in the United Kingdom and to assess how society is likely to handle future genetic information from tests for polygenic multifactorial conditions. DESIGN: Postal questionnaire survey. SUBJECTS: Sample (n=7000) of members from seven British support groups for families with genetic disorders and a representative sample (n=1033) of the general public who answered questions on applying for life insurance as part of an omnibus survey. MAIN OUTCOME MEASURES: Subjects were asked about their experiences with insurers, the medical profession, employers, and social services. Experiences with insurers are reported here. RESULTS: Altogether 33.4% of the study group had problems when applying for life insurance compared with 5% of applicants in the omnibus survey. Thirteen per cent of study respondents from subgroups who represented no adverse actuarial risk on genetic grounds reported that their treatment by insurers seemed to represent unjustified genetic discrimination. CONCLUSIONS: Life insurers may not be operating a consistent policy for assessing genetic information or acting in accord with the actuarial risks brought to them. The inconsistency suggests error rather than a corporate policy of discrimination based on genetic characteristics. Any future proposals for genetic testing for common or multifactorial disorders should be examined carefully. PMID- 9848908 TI - ABC of sexual health: examination of patients with sexual problems. PMID- 9848907 TI - Diet and the prevention of cancer. PMID- 9848909 TI - Screening for diabetes: what are we really doing? PMID- 9848910 TI - Please come back-the patient won't take her cotton wool PMID- 9848911 TI - How to get a grant funded. PMID- 9848912 TI - Colombia: the winner takes all. PMID- 9848913 TI - Hospital at home. Schemes evolve gradually. PMID- 9848914 TI - Acceptability of early discharge, hospital at home schemes. Treatments that can be safely and acceptably managed at home need to be defined. PMID- 9848915 TI - Role of hospitals in NHS must not be undervalued. PMID- 9848916 TI - Combination treatment is rare in patients with rheumatoid arthritis. PMID- 9848917 TI - Research is needed to determine how to integrate complementary medicine into the NHS. PMID- 9848918 TI - Deaths from low dose paracetamol poisoning. Executive action is needed to change national guidelines. PMID- 9848919 TI - Limitation of over the counter sales of paracetamol. Packaging policy is unlikely to achieve its aim of reducing suicide. PMID- 9848920 TI - National patient groups are a resource underutilised by GPs. PMID- 9848921 TI - Postnatal depression. Postnatal depression is not being missed in primary care. PMID- 9848922 TI - National registry is also needed for neurological implants. PMID- 9848923 TI - Inequalities in health. Report on inequalities in health did give priority for steps to be tackled. PMID- 9848924 TI - The Bristol affair. Surgeons were treated unjustly. PMID- 9848925 TI - Undergraduate education must include improving health of minority ethnic communities. PMID- 9848926 TI - Charles david marsden PMID- 9848927 TI - Consultants issue guidance on professional advisory panels PMID- 9848928 TI - Where women have No doctor: A health guide for women PMID- 9848929 TI - An intelligent Person's guide to ethics PMID- 9848930 TI - Conflict in general practice PMID- 9848931 TI - Injury prevention: An international perspective PMID- 9848932 TI - La maladie de sachs PMID- 9848933 TI - Website of the week PMID- 9848934 TI - Quality and the invisible man. PMID- 9848936 TI - The everyday and the soon to be everyday PMID- 9848935 TI - If you want a job doing PMID- 9848938 TI - One third of children in road traffic accidents develop post-traumatic stress disorder PMID- 9848937 TI - Antenatal screening for syphilis should not be abandoned PMID- 9848939 TI - Intranasal corticosteroids should be used for allergic rhinitis PMID- 9848940 TI - Unrecognised infection hinders STD control in africa PMID- 9848941 TI - Insurers discriminate unfairly on basis of genetic information PMID- 9848942 TI - Diabetes screening should address the risk of complications PMID- 9848943 TI - 8th International Congress on Obesity. Paris, France, 29 August-3 September 1998. Abstracts. PMID- 9848944 TI - Secondary photon scatter in imaging cassettes. AB - Secondary photon processes within a cassette image receptor have been identified in the literature for at least three decades. These secondary processes have varying effects on the contrast, resolution and noise of a resulting radiographic image. The magnitude of these effects can be quantified in terms of the percentage of scatter to total absorbed energy for each process recorded by the radiographic cassette, and also as a reduction in system MTF. Two of these processes, cassette face scatter and particularly film crossover, have been subject to considerable past study. The third, due to K-characteristic photon 'crossover', has only recently been quantified. The total magnitude of all secondary cassette effects for a radiographic image, could vary from a worst case where in excess of 40% of the total absorbed energy is due to secondary processes (for an aluminium cassette with Gd2O2S screen with high crossover film, exposed at high kVp) to a best case of less than 10% (for a carbon fibre cassette, tantilate screens with 'zero crossover' film) leading to corresponding contrast reductions of > 40% to < 10% respectively. The MTF components of all secondary effects have been determined and are shown to decrease rapidly with increasing spatial frequency. This results in the MTF of the total image receptor system being reduced for high spatial frequencies by the same percentage as the contrast reduction. Theory predicts that the affected image noise due to cassette secondary processes will be complex and vary with spatial frequency. PMID- 9848945 TI - Monte Carlo simulation of electron cones used in electron beam therapy. AB - A Siemens Mevatron KV2 accelerator installed at the Royal Adelaide Hospital employs cylindrical solid-walled electron cones for some electron collimation. The cones being used at present result in treatment fields that do not always conform with the International Electrotechnical Commission (IEC) Standards (particularly at high energies). The aim of this project was to simulate the existing cones using Monte Carlo methods in order to evaluate potential cone modifications required to overcome the field irregularities. Simulations were performed using the EGS4 (Electron Gamma Shower version 4, distribution II) Monte Carlo code installed on a DEC Alpha workstation at the University of South Australia. To rigorously simulate the existing electron cones it was necessary to also simulate various components within the treatment head of the linear accelerator. Results of simulations for existing cones were found to be consistent with experimental data. (obtained from Royal Adelaide Hospital beam quality assurance measurements). Two proposed changes to the cones were then simulated and the effects of these alterations were assessed. This study has shown how treatment head simulation techniques can be used to assess the changes in dose distribution that result from alterations to the treatment head and accessories. Within practical engineering constraints modifications to an existing electron collimation system were proposed and theoretically evaluated. PMID- 9848946 TI - Bremsstrahlung production from a linac target in the presence of external magnetic fields. AB - A method to increase the photon fluence of the linac is to apply a magnetic field to the bremsstrahlung producing electrons in the target. This field changes the electron paths, and therefore the photon trajectories. In this research Monte Carlo techniques are used to model the effect of three separate magnetic field configurations applied to a tungsten target, on the forward projection of bremsstrahlung created from incident 6 MeV electrons. A radial fluence spectrum is produced for each magnetic field. The maximum increase in fluence is shown to be around 10% for a physically possible field. We conclude that the cost of constructing and installing such magnets (currently) outweighs the small increase in fluence yield. PMID- 9848947 TI - Mathematical modelling of peristaltic transport of a non-Newtonian fluid. AB - The paper considers the phenomena of peristaltic transport of a non-Newtonian fluid represented as a power law fluid. The governing equations are the modified Navier-Stokes equations and the continuity equation in axisymmetric form. A solution is sought in terms of a perturbation series and it is shown the close proximity between analytical and numerical solutions when considering stream functions for various values of the flow behaviour index. PMID- 9848948 TI - Application of fuzzy-classifier system to coronary artery disease and breast cancer. AB - This paper presents an application of a genetic-algorithm-based representation of fuzzy rules for the classification of coronary artery disease data and breast cancer data. The performance of this fuzzy classifier for classification of coronary artery disease and breast cancer data is evaluated. In this study the concept of fuzzy if-then has been applied of rules proposed by Ishibuchi et al. for a multi dimensional data classification problem which leads to higher classification power. The fitness value of each fuzzy if-then rule was determined by the numbers of correctly and wrongly classified training patterns for that rule. The classification power on real world data for coronary artery disease and breast cancer was thus demonstrated by computer simulations. PMID- 9848949 TI - Comparing dose calculation algorithms for an orthovoltage beam in a bone phantom. AB - The aim of this work was to compare dose calculation algorithm results at orthovoltage energies for a phantom composed of a bone slab in water. The calculation methods investigated were: no correction, ETAR, Batho, convolution/superposition and Monte Carlo. All algorithms calculated depth dose curves in a water phantom within 4% of experiment. However in the bone phantom, differences of over 40% between the No Correction/ETAR/Batho/Convolution and Monte Carlo results in the 1 cm thick bone slab were observed. These differences are predominantly because the algorithms do not account for the differing atomic number of the bone compared to water. The increased dose to bone and the tissue adjacent to the bone interface should be considered when treating with orthovoltage photons. PMID- 9848950 TI - Shielding properties of fibre cement wallboard. AB - Transmission data for a fibre cement wallboard (villaboard) are determined for use in diagnostic shielding designs. Villaboard is found to be more attenuating than plasterboard e.g. 9 mm of villaboard is equivalent to 16 mm of plasterboard. PMID- 9848951 TI - Commitment to improving indigenous health. PMID- 9848952 TI - Planning for better health outcomes requires indigenous perspective. PMID- 9848953 TI - Aboriginal health spending: more needed, but where? PMID- 9848954 TI - A 'whole of government' approach needed on indigenous health. PMID- 9848955 TI - At risk in two worlds: injury mortality among indigenous people in the US and Australia, 1990-92. AB - This paper outlines the commonalties and unique differences in injury experience among the indigenous people in the United States and Australia. Injury mortality rates among Indigenous people in the United States and Australia are approximately 2-3 times greater than rates for the non-Indigenous population in each country. Motor vehicle-related injuries accounted for one-third of the injury deaths for Native Americans and Australian Aboriginals. Suicide accounted for more deaths in Native Americans (15.5 per 100,000) than it did for Australian Aboriginals (11.1 per 100,000), whereas the injury death rate in Australian Aboriginals due to poisoning was almost twice that of Native Americans. Culturally appropriate interventions tailored to specific local settings and problems will be necessary to reduce injury mortality among Indigenous people. PMID- 9848956 TI - The contribution of community health surveys to aboriginal health in the 1990s. AB - Community health surveys take place in many Aboriginal communities. We considered these surveys to determine their potential to contribute to Aboriginal health in the 1990s. Community health surveys--also known as health audits, community health screenings or check-ups--usually consist of a team of health professionals travelling to an Aboriginal community to measure a wide variety of parameters on as many of the people in the community as possible. For the individual participant, community health surveys represent a sporadic screening program which should meet the World Health Organization's criteria for screening. From the population health perspective, these surveys represent prevalence surveys which may contribute little new knowledge regarding Aboriginal health and do not, of themselves, change the urgent need for preventive health programs. Community health surveys should meet minimum scientific standards (i.e. have a clearly stated aim and use valid measurements and statistical techniques) and should incorporate practically feasible protocols and services for the follow-up of individuals with screen-detected abnormalities. They must have ethical and community approval and incorporate genuine consultation and feedback of results to the Aboriginal communities involved, in order for them to be justified. PMID- 9848957 TI - Positions and training of the indigenous health workforce. AB - A survey of community-controlled and state health services was conducted in 1995 as part of a needs assessment for a tertiary training initiative in applied epidemiology. Information for 792 Indigenous people in health-related occupations was obtained. Mean time in the current position was 4.8 years, 75% were in designated Aboriginal positions and 44% were health workers. Of the total, 49% had a diploma or certificate, but only 3% had a bachelors degree. The latter compares with 75% of the public health workforce generally which has a bachelors degree or higher. The high proportions of health worker and designated Aboriginal positions, short tenure and low level of degree training suggest that there are a range of employment and training issues which need to be addressed if an appropriate level of Indigenous workforce participation is to be achieved and Indigenous health professionals are to have access to the same vertical and lateral employment mobility as non-Indigenous workers. In particular, there is a huge need to enhance Indigenous participation in health workforce training, and to develop strategies for certification and recognition of the wide range of non course-based training being undertaken. PMID- 9848958 TI - Beneficial impact of the homelands movement on health outcomes in central Australian aborigines. AB - OBJECTIVE: This study compares prevalence of obesity, hypertension and diabetes in two groups of Aboriginal adults: those living in homelands versus centralised communities in central Australia. It also compares weight gain, incidence of diabetes, mortality and hospitalisation rates between the groups over a seven year period. METHODS: Baseline survey of 826 Aboriginal adults in rural central Australian communities in 1987-88 with a follow-up survey of 416 (56% response rate, excluding deaths). Each time, they had a 75 g oral glucose tolerance test (OGTT), and blood pressure and anthropometry measurement. Deaths and hospitalisations for all of the original cohort were recorded for the seven-year period. RESULTS: Homelands residents had a lower baseline prevalence of diabetes (risk ratio [RR] = 0.77, 0.59-1.00), hypertension (RR = 0.66, 0.54-0.80) and overweight/obesity (RR = 0.70, 0.59-0.83). The incidence of diabetes was lower among homelands residents (RR = 0.70, 0.46-1.06). They were less likely to die than those living in centralised communities (RR = 0.56, 0.37-0.85) and less likely to be hospitalised for any cause (RR = 0.79, 0.71-0.87), particularly infections (RR = 0.70, 0.61-0.80), injury involving alcohol (RR = 0.61, 0.47 0.79) and other injury (RR = 0.75, 0.60-0.93). Mean age at death was 58 and 48 years for residents of homelands and centralised communities respectively. CONCLUSION: Aboriginal people who live in homelands communities appear to have more favourable health outcomes with respect to mortality, hospitalisation, hypertension, diabetes and injury, than those living in more centralised settlements in Central Australia. These effects are most marked among younger adults. PMID- 9848959 TI - Improving cervical screening in a remote aboriginal community. AB - Improved cervical screening has been identified as a priority in reducing the incidence of cervical cancer among Aboriginal women. This paper discusses the cervical screening recommendations of a women's health project developed by Nganampa Health Council (NHC), the Aboriginal-controlled medical service meeting the health needs of the people of the Anangu Pitjantjatjara (AP) Lands. A community participation public health model was used in program development. A process including community and staff consultation, literature review and prioritisation informed the program development, ensuring strategies thereby derived were culturally acceptable as well as practical and scientifically valid. The epidemiology of cervical cancer and screening in this community is discussed. Strategies aimed at maximising patient participation, improving patient satisfaction, running an effective call and recall system, improving quality of cervical smears, improving management of women with abnormal smears and evaluating the program are suggested. The involvement of the community in a scientifically sound, public health approach may increase the likelihood that the strategies suggested will be effective in this community. Both the model of program development and the cervical screening strategies derived are likely to be of relevance for other Aboriginal communities. PMID- 9848960 TI - The vaccination status of aboriginal and Torres Strait Island children in far north Queensland. AB - A survey was undertaken to ascertain the vaccination status of all 773 Aboriginal and Torres Strait Island children who were born and remained in Far North Queensland during the 1993-94 financial year. Only 42% had received all 15 vaccines scheduled in the first two years of life by their second birthday. More who resided in remote communities were fully vaccinated (64%) by then than those who lived in rural towns (32%) or an urban setting (21%) (p < 0.01). The 445 children who were not fully vaccinated required a median of three vaccines to have been fully vaccinated by the second birthday. Of these, 146 (33%) required only one vaccine, nearly 60% of whom would have been fully vaccinated if they had had the fourth (18-month) dose of diphtheria-tetanus-pertussis (DTP) vaccine. Of the 445, 143 (32%) required five or more vaccines to have been fully vaccinated. Only 26% and 36% of the children received all the vaccines scheduled at six and 12 months of age, respectively, on the same day. However, the eventual uptakes of the three vaccines scheduled at six months of age were very similar (approximately 80%) and simultaneous vaccination with the two vaccines scheduled at 12 months of age would have made a very limited (approximately 4 percentage points) impact on the overall percentage of fully vaccinated children. Considerably more than simple and apparently logical strategies will be required to ensure that Indigenous children in Far North Queensland are adequately vaccinated. A systematic approach, with a careful understanding of the barriers to routine vaccination and a means of prospectively tracking the vaccination status of each child, will be needed if state and national vaccination goals are to be met. PMID- 9848961 TI - Seasonality of low birthweight in indigenous Australians: an increase in pre-term birth or intrauterine growth retardation? AB - We have analysed birthweights of 4,508 Aboriginal and Torres Strait Islander livebirths in the Kimberley region of Western Australia from 1981-93. Mean birthweight varied significantly according to month of birth (F(11) = 2.57, p = 0.003) and low birthweight babies were more common during the wet season. A significant increase in the proportion of very low birthweight (VLBW) babies was observed during the wet season compared with the dry season (OR 2.73; 95% CI 2.3 3.67; p < 0.001); whereas babies weighing 1,500-2,499 g were not significantly more common during the wet season (OR 1.06; 95% CI 0.96-1.17; p = ns). The results indicate that adverse environmental conditions may be associated with increased risk of VLBW. Since newborns weighing less than 1500 g are very likely to be pre-term (< 37 weeks' gestation), the findings also suggest that seasonality of birthweight may be due to an increase in pre-term births rather than an increase in intrauterine growth retardation. Further research is required to identify the underlying causes of an increase in VLBW babies during the wet season. PMID- 9848962 TI - Hospital use for potentially preventable conditions in aboriginal and Torres Strait Islander and other Australian populations. AB - The poor state of Aboriginal and Torres Strait Islander health has been documented in many ways, most obviously by comparing the relatively higher age specific mortality and morbidity rates. This paper demonstrates the use of acute hospital separation data as a way to identify potential deficiencies in providing appropriate primary health care services for Aboriginal and Torres Strait Islander populations. It does so by using 'ambulatory sensitive conditions': those conditions (and procedures) for which high-quality appropriate primary health services deliverable under ideal circumstances are though to potentially reduce or eliminate the need for hospitalisation. Potential or realised access to primary care is not analysed directly using primary health service data. In this study, 1993-94 acute hospital separation data from NSW, Queensland, South Australia, Western Australia and the Northern Territory were used to calculate separation rates and odds ratios for Aboriginal and Torres Strait Islander and non-Aboriginal and Torres Strait Islander populations. Age-specific acute hospital separation rates for ambulatory sensitive conditions were 1.7 to 11 times higher for the Aboriginal and Torres Strait Islander populations studied. This supports clinical contentions that much Aboriginal and Torres Strait Islander morbidity and mortality is preventable and that further consideration is needed to service delivery reform at all levels in the health system and the distribution of funding. PMID- 9848963 TI - Out of sight, out of mind? Licensed clubs in remote aboriginal communities. AB - Most Aboriginal communities in the Northern Territory have used provisions under the NT Liquor Act to impose total or partial bans on alcohol and only eight (as of 30 June 1995) had established licensed clubs. This article examines (a) patterns of alcohol consumption in communities with clubs, and (b) economic, social and political aspects of the place of clubs in these communities. Alcohol consumption was estimated on the basis of 'purchase into store' figures for 1994 95 provided by the NT Liquor Commission. Among male drinkers, consumption of absolute alcohol in 1994-95 was estimated at 42.5 litres a head, 76% higher than the figure for the NT as a whole, which in turn was 42% above the national level. Similar differences were found for female drinkers. Combined retail turnover of the seven clubs that traded throughout the year was estimated at $8.1 million. It is argued that many clubs are able to use their monopolistic control of a valued resource to become powerful political institutions in communities, sometimes leaving non-drinkers effectively disenfranchised. It is also argued that the health consequences of these high consumption levels have received far less attention than the effects of Aboriginal public drunkenness in urban areas. It is concluded that, while the rights of Aboriginal communities to establish community controlled clubs should be respected, the notion that they are under some sort of obligation to do so should be exposed as a measure likely to add to the health burdens of people already inadequately served by health, education and other services. PMID- 9848964 TI - School screening in remote aboriginal communities--results of an evaluation. AB - As part of ongoing quality assurance, the effectiveness of the school screening program in meeting the health needs of Aboriginal children in a rural district in the Northern Territory Top End was evaluated. The major health problems of Aboriginal children were analysed for their suitability for screening programs. A prospective cross-sectional study used routinely collected field data from the school screening program in 1993. To ascertain follow-up, children who had failed screening tests had their clinic notes reviewed. A total of 774 children from 11 remote communities were screened. The results confirmed high level of disease, with rates for anaemia, malnutrition and trachoma reaching 39%, 22% and 26% respectively. Nearly one-third failed the hearing screening, urinalysis was abnormal in 19%, 3% failed visual acuity and 6% were considered to have abnormal heart auscultation. Many of the major health problems did not meet the recommended criteria for screening programs and others would be better dealt with by ongoing surveillance rather than a single screening. Some abnormalities found on screening were inadequately investigated and/or treated. This evaluation has demonstrated a limited role for school screening in identifying and meeting the health needs of Aboriginal children living in remote areas. In collaboration with the NT Department of Education, a school-age child health policy, including a new school-age child health surveillance program, is being developed. This promotes greater participation by communities, families and schools with the aim of improving the health and learning outcomes for all school-age children in the NT. PMID- 9848965 TI - Differences in growth among remote and town-dwelling aboriginal children in the Kimberley region of Western Australia. AB - This paper documents the growth of Aboriginal children in remote communities and gazetted towns of the Kimberley region in the far north of Western Australia over the past 20 years. The study's specific aim was to compare the height and weight growth of children in different environments from birth to five years of age. From 1979 to 1983, children living in towns were significantly taller and heavier than their counterparts in remote communities. From 1984 to 1988 and thereafter, there were no significant differences in the growth patterns of children in towns and remote communities. Overall, weight-for-age and height-for-age of children in remote communities has improved since the 1970s relative to their town-dwelling counterparts. These changes in growth shed light on the quality of the social and physical environment over the past 20 years and may assist with the development of future programs for child health. PMID- 9848966 TI - Washing machine usage in remote aboriginal communities. AB - The use of washing machines was investigated in two remote Aboriginal communities in the Anangu Pitjantjatjara homelands. The aim was to look both at machine reliability and to investigate the health aspect of washing clothes. A total of 39 machines were inspected for wear and component reliability every three months over a one-year period. Of these, 10 machines were monitored in detail for water consumption, hours of use and cycles of operation. The machines monitored were Speed Queen model EA2011 (7 kg washing load) commercial units. The field survey results suggested a high rate of operation of the machines with an average of around 1,100 washing cycles per year (range 150 and 2,300 cycles per year). The results were compared with available figures for the average Australian household. A literature survey, to ascertain the health outcomes relating to washing clothes and bedding, confirmed that washing machines are efficient at removal of bacteria from clothes and bedding but suggested that recontamination of clothing after washing often negated the prior removal. High temperature washing (> 60 degrees C) appeared to be advantageous from a health perspective. With regards to larger organisms, while dust mites and body lice transmission between people would probably be decreased by washing clothes, scabies appeared to be mainly transmitted by body contact and thus transmission would be only marginally decreased by the use of washing machines. PMID- 9848967 TI - Private business: the uptake of confidential HIV testing in remote aboriginal communities on the Anangu Pitjantjatjara Lands. AB - BACKGROUND AND METHODS: Despite a concentration of risk factors for HIV transmission, many remote Aboriginal communities in central Australia have a low uptake of HIV testing. We studied the uptake of HIV testing in six clinics in remote Aboriginal communities following the introduction of voluntary confidential testing to assess the impact of the intervention and to determine if the program was reaching people most at risk of HIV infection and transmission. SETTING: The study was conducted by Nganampa Health Council, an Aboriginal controlled health service on the Anangu Pitjantjatjara Lands in the far north west of South Australia. RESULTS: Since the introduction of confidential coded testing in August 1994 the number of HIV tests provided through the remote clinics has increased from 83 tests/year to 592 tests/year. In the 12-month audit period (August 1, 1995, to July 31, 1996) 62.7% of women aged 20-24 years, 44.6% of people aged 12-40 years and 24% of the total population had an HIV test. Fifty per cent of tests were accounted for by the 15-25 year age groups and 60% of tests related to an STD consult. DISCUSSION: This study shows that a high uptake of HIV testing in high-risk groups can be achieved in remote Aboriginal communities where a high level of confidentiality is maintained. PMID- 9848968 TI - Age at first episode of venereal syphilis in an aboriginal population: an application of survival analysis. AB - OBJECTIVE: To identify the age at which screening for syphilis could be ceased in Kimberley Aboriginal people who had remained seronegative and whether this age was the same for people with and without a past history of gonorrhea. DESIGN: Retrospective cohort study. SETTING: The Kimberley region in the far north of Western Australia. PARTICIPANTS: Kimberley Aboriginal residents. MAIN OUTCOME MEASURE: Age at seroconversion. RESULTS: Survival analysis showed that seroconversion due to venereal syphilis occurred largely during the years between the late teens and early 20s. The probability of seroconverting by a given age was related to the time at which the subject's last gonorrhea infection occurred. CONCLUSIONS: As a result of this study, annual periodic screening for syphilis is now offered to Kimberley Aboriginal people only between the ages of 15 and 25 years, allowing for the redistribution of resources to other activities that have greater potential to decrease the incidence of all STDs. PMID- 9848969 TI - Leprosy control in the Northern Territory. AB - An epidemic of leprosy occurred among Aboriginal people of the Top End of the Northern Territory following its introduction towards the end of the 19th Century. The extent of this outbreak became apparent through community surveys conducted in the 1950s which revealed that one in 10 Aboriginal people in some areas were affected by leprosy. Initial control activities were outbreak-focused, directed at case finding and management. Case finding was by systematic community survey. Case management included appropriate rehabilitation and reconstructive surgery. Regular review of treated patients ensured early detection of relapse and detection and treatment of sequelae. Education and full participation of Aboriginal health workers in the diagnosis and management of cases provided local expertise at the hospital and community level. The case detection rate fell from 270 per 100,000 in the Aboriginal population in 1951 to four per 100,000 in 1997. Elimination of transmission is now the objective of the control program. Combining of the tuberculosis and leprosy control activities of the Territory Health Service in 1996 resulted in increased efficiency of the mycobacterial services. PMID- 9848970 TI - Restriction of the hours of sale of alcohol in a small community: a beneficial impact. AB - The population of Halls Creek, a small town in the remote Kimberley region of Western Australia, is predominantly Aboriginal. After many years of high alcohol consumption, a number of measures were taken in an effort to redress its negative influence on the community. Key among these was a restriction on the trading hours when 'take-away' alcohol was available. The effects of this intervention were monitored by examining longitudinal patterns of alcohol consumption, incidence of crime and outpatient data at the local hospital. The data were compared with equivalent periods prior to the restricted trading hours. A decrease in alcohol consumption was observed for each of the two years following the intervention. Overall, incidence of crime declined. Alcohol-related presentations to the hospital and presentations resulting from domestic violence decreased relative to the equivalent quarterly period prior to the intervention. There were short-term fluctuations observed, particularly with domestic violence, where presentations (of lesser severity) became more frequent during several quarters. Emergency evacuations as a result of injury showed a marked decrease. The consistency of trends across a variety of health and social data show a positive effect after the implementation of restricted trading hours. While a direct effect is likely, a multitude of concurrent programs which promote health in the community place limitations on this conclusion. The process in achieving change, supported by statutory measures, has, however, been successful in curbing the morbidity and mortality experienced by the community. PMID- 9848971 TI - Pap smear screening at an urban aboriginal health service: report of a practice audit and an evaluation of recruitment strategies. AB - A culturally appropriate women's health service was established at an Aboriginal community-controlled health service in Darwin in 1994. An initial file audit found that 48% of included women had ever been screened with a Pap smear and 37% of women were considered to have been adequately screened. The enhancement of opportunistic screening by file tagging had a modest effect on screening coverage over a 12-month period for women who attended the health service. The proportion of these women who were adequately screened increased from 43% to 48% and of those ever screened increased from 54% to 62%. A randomised trial of recruitment interventions including personal approach, letter and control groups was subsequently performed for women for whom Pap smears were overdue or not recorded. The impact of both interventions on the number of Pap smears performed was low, with 7% of women in the personal approach group, 2% of women in the letter group and no women in the control group having Pap smears during the three month follow-up period. Low rates of abnormalities were observed for women having Pap smears over a two-year period. The minimal effect of a formal reminder system and letters at this urban Aboriginal health service has resulted in a re orientation of activities towards strengthening opportunistic screening and the continued promotion of Pap smears in a range of clinic and community settings. It is important to place Pap smear screening in the context of other social, economic and health priorities for Aboriginal women and health workers. PMID- 9848972 TI - Aboriginal perspectives of diabetes in a remote community in the Northern Territory. AB - This study explores the knowledge and beliefs of diabetes in a group of Aboriginal people from a remote community in the Northern Territory. Information was gathered from participants through a combination of group discussions, semi structured interviews and informal conversations. The four themes of explanation of diabetes were: worry, food, family and infections. The most common means of preventing or treating diabetes were stopping worry and changing one's diet. Significantly, few of the participants believed that medication was effective, and weight loss and exercise were not mentioned. The results highlight the miscommunication about diabetes that has occurred in this population. For effective communication and management of diabetes, health information must incorporate the contemporary health beliefs of the clients. PMID- 9848973 TI - Measles vaccine effectiveness in central Australian aboriginal children vaccinated at or after eight months of age. AB - An outbreak of measles in central Australia in 1994 provided the first opportunity to evaluate the effectiveness of the measles vaccine given to Aboriginal children at nine months of age since 1984. Children eligible for the study that was conducted in one community in the region were aged between nine months and 10 years. Eight of the 109 eligible children developed measles. The only unvaccinated child also developed measles. Vaccination failures occurred in 7.8% (6/77) of children vaccinated between eight and 11 months of age and in 3.2% (1/31) vaccinated after 11 months of age. Overall vaccine effectiveness was 93.5%. The level of vaccine uptake in central Australia is high and the last region-wide outbreak before 1994 occurred in 1981-82. If the age of vaccination against measles is to be determined by the average age of infection; the age of vaccination should now be raised to 12 months of age; this is the age at which Aboriginal children in all other states and all children in Australia are vaccinated. PMID- 9848974 TI - Indigenous health: a special moral imperative. AB - The provision of health services to Indigenous people is not perceived by many Australians to be a moral issue. Indigenous health, however, is not only a moral issue, it is a moral issue that deserves special consideration. In many sectors of society, the correct moral path is unclear, but the circumstances of Indigenous health warrant special consideration which policy makers and health care administrators are uniquely placed to render. The setting of Australia was at the expense of Indigenous flourishing. There is little doubt that many of the current poor health outcomes of Indigenous Australians result from their past impoverishment. We argue that each member of Australian society has inherited a collective moral responsibility, along with the social assets accrued at the expense of Indigenous Australians, irrespective of their personal complicity. Government, as representatives of the people, has a responsibility to repay some of this society's accrued moral debt through the allocation of resources independent of issues of equity. PMID- 9848975 TI - Postmodernism and the democratisation of public health. PMID- 9848976 TI - Do media help or harm public health? PMID- 9848977 TI - Wellbeing in a New Zealand secondary college--an evaluation. PMID- 9848978 TI - Arthroscopic management of temporomandibular closed lock. AB - Sixty-three consecutive patients with painful closed lock involving 83 temporomandibular joints were treated by surgery and followed up for at least 24 months. Prior to surgical treatment all had imaging to demonstrate that the cause of their painful jaw locking was an anterior displaced disc without reduction. All patients had received at least three months non-surgical treatment without response. The arthroscopic surgery consisted of release of adhesions, manipulation and placement of 25 mg hyaluronic acid into the joint space. No significant surgical complications occurred. Postoperative treatment involved physical therapy and stabilization splints. At 24 months after surgery 57 patients (90.5 per cent) had no pain and month opening had improved from preoperative 27.2 +/- 5.4 mm to 44.4 +/- 4.1 mm (p < 0.001). This study shows that arthroscopic surgery with continuing non-surgical therapy is a highly effective treatment for painful closed lock. PMID- 9848979 TI - Focal epithelial hyperplasia in an HIV positive man. An illustrated case and review of the literature. AB - Focal epithelial hyperplasia (FEH) is a rare but distinctive entity of viral aetiology with characteristic clinical and histopathological features. It is usually seen in children and adolescents of American Indian and Eskimo background; however, it has recently been described in adults who are HIV positive. Suppression of the immune system leaves the individual vulnerable to opportunistic infections. With improved management of immunocompromised patients it is increasingly possible that the general dental practitioner will encounter secondary oral infections such as FEH and must therefore be able to recognize, diagnose and treat these lesions. The following report presents a case of FEH in an HIV-infected man, and is followed by a review of the literature. PMID- 9848980 TI - Sublingual hypoglossal neurilemmoma. Case report. AB - Neurilemmomas of the peripheral segment of the hypoglossal nerve are extremely uncommon neoplasms. A rare case of hypoglossal neurilemmoma located in the sublingual space is reported and the differential diagnosis of sublingual mass lesions is discussed. PMID- 9848981 TI - Comparison of health behaviour and oral/medical conditions in non-insulin dependent (type II) diabetics and non-diabetics. AB - One hundred and two dentate patients with type II diabetes mellitus and 98 non diabetic subjects were examined for oral conditions and metabolic state. Self reported health behaviour was analysed. From factor analysis four factors emerged: general health behaviour (GHB), perceived fatigue (PF), diet control (DC) and regular diet (RD). In diabetics PF, DC and RD were significantly higher than that in non-diabetics. Patients with diabetes were more likely to control their disease through a programme of decreased kilojoule intake leading to weight management. However, they tended to tire. The mean gingivitis index was significantly higher (p < 0.01) among diabetics (2.39) than among non-diabetics (1.99). The number of missing teeth was significantly higher (p < 0.01) for diabetics (6.7) when compared with non-diabetics (4.3). On the other hand, aetiological factors (plaque, calculus) and the level of dental health behaviour as expressed in the HU-DBI scores were similar. Probing pocket depth did not differ statistically between groups. The increasing number of missing teeth in diabetics may primarily result from severe periodontitis with tooth mobility or deep pockets. Findings in this study suggest that the difference in the severity of periodontitis between diabetics and non-diabetics was significant although aetiological factors and the level of dental health behaviour were similar. PMID- 9848982 TI - Dermal psoriasis involving an oral split-skin graft. Case report. AB - Psoriasis is a chronic inflammatory skin disease which very rarely affects the oral cavity. It has many forms of expression on cutaneous and mucosal surfaces. This paper describes an unusual case of psoriasis with oral involvement affecting the skin graft used to reconstruct an oral mucosal defect. Diagnosis was based on clinical and microscopic findings and confirmed by the concurrent cutaneous disease. PMID- 9848983 TI - Idiopathic swelling of the lower lip associated with topical anaesthesia. Report of three cases. AB - Three cases of idiopathic swelling of the lower lip are presented. Each developed following administration of a topical anaesthetic agent with unusual clinical manifestations closely resembling allergic angio-oedema. The possibility of psychogenically mediated reactions was considered the contributing aetiological factor which showed as a somatic manifestation in the lower lips. PMID- 9848984 TI - Dens evaginatus on a wisdom tooth: a diagnostic dilemma. Case report. AB - Pericoronitis is the most common odontogenic problem associated with the wisdom tooth in young men and women. Patients may present with problems associated with infection, swelling and pain. However, other associated pathology such as caries, periodontal disease and referred pain from the temporomandibular joint must be investigated when treating pain from the wisdom tooth. The authors wish to present a case in which the pain from a wisdom tooth was due to a fractured dens evaginatus. The importance of this dental anomaly should not be overlooked. PMID- 9848985 TI - Service provision trends between 1983-84 and 1993-94 in Australian private general practice. AB - Patterns of service provision are expected to change over time, reflecting the dynamics of factors such as oral health status and trends in population demographics. The aim of this study was to identify trends in service provision over time. Data were collected from a random sample of Australian dentists in 1983-84, 1988-89, and 1993-94. Changes observed between 1983-84 and 1993-94 included a trend towards increased proportions of patients in older age groups, decreases in the percentage of persons receiving restorative care (from 40.2 per cent to 37.7 per cent) and prosthodontic care (9.3 per cent to 8.0 per cent), and increases in diagnostic (from 38.7 per cent to 46.0 per cent), preventive (20.9 per cent to 25.2 per cent), endodontic (5.3 per cent to 7.1 per cent), and crown and bridge services (3.1 per cent to 4.7 per cent) per visit. Other services such as orthodontic and general/miscellaneous services showed increases over the first half of the study period, but these did not continue between 1988-89 and 1993-94. The total number of services per visit increased over the study period for adult patients. These findings indicate changing patterns of practice over time, consistent with an increasing orientation towards prevention of disease and maintenance of a natural dentition. PMID- 9848986 TI - An assessment of attitudes to, and extent of, the practice of denture marking in South Australia. AB - Denture marking or labelling is not a new concept in either prosthetic or forensic dentistry and its routine practice has been urged by forensic odontologists internationally for many years. In the general community it is often recommended for institutionalized persons to prevent confusion of ownership of dentures. In Australia, the Nursing Home Standards require that dentures of residents be 'discreetly labelled' and marking of all dentures is recommended by the Australian Dental Association. In some countries the marking of dentures is regulated by legislation, but elsewhere there seems to be a reluctance to effect this practice. Various methods which have been proposed include the insertion of an identifying label during the fabrication of the dentures with the utilization of a number of materials and coding systems. This study reports the results of a survey undertaken to determine the extent of the practice of denture marking in South Australia, the methods in use, and the attitudes of dentists, dental technicians and institutions to it. The results indicated that 24.5 per cent of all practitioners providing removable prostheses to their patients include an identifying label as part of the service on some occasions. This included 19.9 per cent of general dental practitioners, 25 per cent of specialist prosthodontists, 57.1 per cent of practitioners with training in forensic odontology, and 43.5 per cent of clinical dental technicians. No practitioner labelled dentures routinely. Reasons cited for not labelling dentures included cost, lack of awareness of standards and recommendations and a belief that it was of little importance. PMID- 9848987 TI - Handpiece degradation associated with performance testing of diamond rotary cutting instruments. AB - Reliable standardized testing of dental rotary cutting instruments remains a problem because of the many significant parameters that require identifying and quantifying. In a previous study, handpiece wear or degradation was identified as a potentially significant confounding variable leading to the suggestion that the use of industrial handpieces might be necessary. Therefore, the objective of this investigation was to measure and compare industrial and dental air rotor degradation during a simulated diamond rotary cutting instrument test. Two dental and two industrial products were tested in a random sequence using three replicate samples of each air rotor. The simulated test consisted of 10 diamond instruments each making 90 passes through a float glass substrate. The testing apparatus comprised a computer-interfaced device that controlled the contact force between the cutting diamond and the substrate. Air rotor performance was based on the time required to cut a standardized path through the substrate. The performance change that occurred from diamond instrument to instrument during the simulated test was attributed to air rotor degradation. Multiple regression analyses showed significant differences between replicate samples of the same type and between regression intercepts for different air rotor types. Surprisingly, four air rotors exhibited improving performance during the simulated test. Changing air rotor performance must be considered to achieve reliability for the testing of diamond rotary cutting instruments. PMID- 9848988 TI - The effect of cleaning on blood contamination in the dental surgery following periodontal procedures. AB - Blood contamination of 16 surfaces in the dental surgery was investigated using the Kastle-Meyer test for haemoglobin, after three types of periodontal procedures had been performed on a total of 30 patients. The effect of cleaning surfaces contaminated by blood was investigated using the same test. Cleaning materials used in the dental surgery were tested to rule out the possibility of false positive outcomes and the sensitivity of the test was determined prior to the study. The results show a marked variation in the degree of contamination and efficacy of cleaning following treatment. Overall, root planing was associated with the most widespread and frequent blood contamination and gingival surgery the least. The surgery work surface, edge of the spittoon, aspirator tube and ultrasonic scaler handpiece into which the ultrasonic insert fits, were the most frequently contaminated surfaces. The work surface, dentist's pen, light switch and handle were cleaned most effectively. The least effectively cleaned surfaces were the water dispenser switch, aspirator tube, bracket table and ultrasonic scaler handpiece. Methods for reducing this potential source of cross-infection are discussed. PMID- 9848989 TI - The changing role of dental auxiliaries: a literature review. AB - A review of the literature on dental auxiliaries and their utilization in the dental workforce is presented, and their future role is examined in the light of broader issues relating to changing disease patterns and service delivery. Legislative factors, productivity, quality assurance and the potential scope for dental auxiliaries in both the public sector and private practice setting are discussed. A strategy for adopting a team approach in the delivery of dental services is suggested, which would necessitate the dentist taking on the role of team leader and maintaining responsibility for overall treatment planning and quality assurance. Dental auxiliaries could provide basic preventive and restorative dental services, allowing dentists to concentrate on providing more complex high-technology treatment. Implications for the future training of dental auxiliaries are presented. PMID- 9848990 TI - The effects of early orthodontic force application on pulp test responses. AB - The effects of orthodontic tooth movement on pulpal responses are of interest to the clinician. Alterations to pulpal physiology may result in altered responses to external stimuli. This pilot project tested a small group of orthodontic patients during the early stages of treatment. Heat, cold and electrical stimuli were applied prior to treatment, after the placement of fixed appliances and after one month of force application. A smaller group was tested after two months of force application. One month after force application there was a lack of response to electrical stimulation but there was continued response to thermal stimulation. The lack of response to electrical stimulation continued in the smaller group up to two months. The results suggest that the outcome of electric pulp testing during orthodontic treatment should be interpreted cautiously. Thermal testing will offer more reliable data. PMID- 9848991 TI - Comparison of exposed dentinal surfaces resulting from abrasion and erosion. AB - The aim of this study was to compare the shape of exposed dentinal surfaces caused by abrasion and erosion with a view to developing a diagnostic clinical test. The study material consisted of 80 natural teeth and 129 dental models obtained from Australian Aborigines known to display considerable dental abrasion due to their diet, and dental models of 37 Caucasians diagnosed with dental erosion through detailed history and dietary analysis. Polyvinyl siloxane impressions were obtained of all occlusal surfaces with dentinal scooping in both the 'abrasion' and 'erosion' groups. All impressions were sectioned buccolingually through the deepest point of the scooped dentine, and then the profiles were photocopied at x2 magnification. The breadth and depth of dentinal profiles were measured to an accuracy of 0.1 mm, enabling ratios of depth:breadth to be determined, and the position of the deepest part of each scooped surface was recorded. The mean depth:breadth ratio of scooped dentine was significantly greater in the Aboriginal natural teeth (0.19 +/- 0.06, mean +/- SE) than in the Aboriginal dental models (0.15 +/- 0.04). Both Aboriginal natural teeth and models with abrasion showed significantly smaller ratios (p < 0.05) than the Caucasian models showing erosion (0.33 +/- 0.07). Furthermore, in the abrasion samples, the deepest region of the scooped dentine tended to be lingually placed more often in maxillary teeth but buccally placed more often in mandibular teeth (p < 0.05). These results indicate that scooped dentine on abraded occlusal surfaces of teeth displays significant differences in shape compared with that caused mainly by erosion. PMID- 9848992 TI - Care, courtesy, consideration, communication. PMID- 9848993 TI - Changes, changes, changes. PMID- 9848994 TI - Toothbrush contamination. PMID- 9848995 TI - Inherited predisposition to colon cancer. AB - Colorectal cancer is one of the most common malignancies in the United States. Although both genetic and environmental factors play a role in colorectal tumorigenesis, recent advances in genetics have more clearly defined the impact of inheritance in the multistep process of the disease. Researchers have identified single genes that confer a susceptibility to familial adenomatous polyposis (FAP) and hereditary nonpolyposis colorectal cancer (HNPCC). Because these genes are inherited in an autosomal dominant fashion, offspring of carriers have a 50% chance of inheriting the gene mutation and its associated risk. The FAP gene, when mutated, initiates the neoplastic process. HNPCC gene mutations disrupt mismatch repair, thus inducing progression of tumor formation. Discovery of these genes has helped our understanding of sporadic colon cancer as well. Genetic testing for the FAP and HNPCC genes is now available, and results of this testing have implications for surveillance and management. In addition, testing raises complex psychosocial and ethical issues. At present, genetic testing is primarily conducted in the research setting, but it will soon be available in the clinical arena. To prepare for the challenges that these new advances will present, nurses must begin now to enhance their knowledge of genetics and its application to oncology. PMID- 9848996 TI - Acute oral pain and mucositis in bone marrow transplant and leukemia patients: data from a pilot study. AB - The purposes of this prospective, repeated-measures descriptive pilot study were to describe patterns of acute oral pain and mucositis in patients receiving a bone marrow transplant or high-dose chemotherapy for leukemia, and to test procedures and instruments before initiating a larger intervention study. A nonprobability, purposive selection process was used to enroll 18 patients admitted to two acute care inpatient hospital units for bone marrow transplantation or leukemia therapy at a university health sciences center in the southeastern United States. Data were collected at baseline, then daily through patient interviews, oral examination, and chart review for at least 3 weeks or until discharge. Research variables were pain intensity, intolerable pain, verbal descriptors of pain, pain relief, and use of pain relief strategies (Pain Assessment Form), mucositis (erythema and ulceration) in eight anatomic locations of the oral cavity (Oral Mucositis Index), voice/talking (Oral Assessment Guide), and mood states (11-item Brief Profile of Mood States). Mild to moderate pain occurred in nearly 70% of patients and was described as "tender," "irritating," and "sore." Patients used pain medicines, mouth care, and mental and physical activities to relieve pain, and reported partial overall relief of pain. Mucositis was mild, with the tongue and buccal and labial mucosa most commonly affected with erythema and the buccal mucosa with ulceration. Voice/talking were only mildly impaired, and mood disturbance was mild. Patterns of pain, mucositis, and mood disturbance were consistent with each other and followed the trajectory described in previous research. Results suggest that nurses should continue to assess these symptoms vigorously and assist patients in selecting multiple management strategies. Research using repeated-measures designs in this acutely ill inpatient population is challenging and needs careful attention by researchers. The results have been used to improve the ongoing larger intervention study. PMID- 9848997 TI - Oncology nursing research priorities: a Canadian perspective. AB - The purpose of the study was to determine oncology nursing research priorities among Canadian oncology nurses. The following two groups of nurses were mailed survey questionnaires: oncology nurse researchers and nurses working in oncology clinical settings who were members of the Canadian Association of Nurses in Oncology (CANO). The questionnaire was based on prior Oncology Nursing Society (ONS) research surveys and the oncology nursing literature. It instructed participants to rank from a list of topics, the five they perceived as research priorities. The response rate was 31% (249/810) for CANO members and 41% (32/78) for nurse researchers. The top five research priorities for CANO members were communication, symptom management, quality of life, pain control/management, and standards of practice. The top five research priorities for nurse researchers were pain control/management, family issues, patient participation in decision making, symptom management, and communication. Many of the same topics appeared in the priority lists of both the CANO members and the nurse researchers. However, the rank ordering of topics was not identical. The lists of CANO members contained both clinical and professional issues, whereas the researchers' lists focused primarily on clinical issues. The findings provide the basis for developing a Canadian national oncology nursing research agenda. In addition, the findings provide direction for practice and education strategic plans as well as information to guide decision making around research funding. PMID- 9848998 TI - Exploring empathy as a variable in the evaluation of professional development programs for palliative care nurses. AB - Research indicates that empathy, a quality regarded as fundamentally important to nursing practice, is a teachable skill. Because empathic nurse-patient relationships are particularly important in the care of the terminally ill, this has direct relevance to the professional development of palliative care nurses. This article discusses the place of empathy as a criterion variable in the evaluation of a professional development program for palliative care nurses introduced at the Centre for Mental Health Nursing Research at Queensland University of Technology, Brisbane, Australia. A modified version of the Staff Patient Interaction Response Scale (SPIRS) was used as a pre- and postintervention measure to assess the expressed empathy of the participating nurses. The modifications to SPIR and its coding system to make it suitable for palliative care nursing, and the mechanisms for improving and evaluating the reliability of this instrument will be discussed. The full description of this particular modification of SPIRS for palliative care research is provided as an example of how this instrument could be used in projects for which nurses undertake the difficult task of providing compassionate care to the terminally ill. PMID- 9848999 TI - Quality of life in patients with endocrine tumors of the gastrointestinal tract: patient and staff perceptions. AB - Patient and staff perceptions regarding the importance of selected quality-of life (QoL) aspects and patient ratings of their present QoL (EORTC QLQ-C30) were investigated. The study comprised 17 patients with endocrine tumors of the gastrointestinal tract and their staff, paired in dyads. Both patients and staff rate physical aspects of life (e.g., "wash yourself, dress, eat, etc.," "have a good health," "not have pain") as most important for experiencing a good QoL. Staff considered work as more important for experiencing a good QoL than did patients. Patients rated their QoL as relatively good and were more satisfied with certain QoL aspects than staff perceived them to be. Staff did not accurately judge a certain patient's satisfaction with various QoL aspects, which suggests that there is room for staff to improve their understanding of these aspects. There were no significant mean value differences between patient and staff ratings of patient anxiety and depression, but no significant correlations within dyads. This suggests that the communication between patients and staff could be improved in this respect. PMID- 9849000 TI - A narrative analysis: a black woman's perceptions of breast cancer risks and early breast cancer detection. AB - The oncology nurse's role in breast cancer management is enhanced by knowledge of the patient's perceptions of risks. This case study elucidates the process by which perceived risks of breast cancer are embedded in sequences of biographic experiences including childhood sexual abuse, childhood injuries, and an abusive marriage. The case study shows that risk perceptions and subsequent delayed breast cancer detection is related to (a) a belief that breast cancer results from "bad luck, or fate"; (b) lack of cancer-related symptoms; (c) belief that a higher power determines ill health; (d) reluctance to turn to others for help while in an abusive marriage; (e) family history of cancer invulnerability since generations of family members died of diabetes, heart disease, and pregnancy related illnesses; and (f) fear of gynecologic exams resulting from childhood sexual abuse. Furthermore, nonapplicability of traditional breast cancer risk factors such as heredity, age older than 30 years at first full-term pregnancy, early menarche, and late menopause prohibit an accurate assessment of self-risk. This case study suggests that breast cancer risk perception often differs from that of biomedical factors, and that an understanding of risk judgments is essential for appropriate therapeutic responses. PMID- 9849001 TI - Narratives of breast symptom discovery and cancer diagnosis: psychologic risk for advanced cancer at diagnosis. AB - In spite of cancer screening programs, women continue to present with advanced breast cancer. How do women decide whether and when to seek an evaluation for self-discovered symptoms? This study examined 104 narratives told by 80 Anglo-, Latina-, and African-American women who participated in 1 of 16 community-based focus groups. The women's narratives contained powerful thematic messages about breast cancer and their expected behavior in the event of a self-discovered breast symptom. Narrative explanations that predicted an increased likelihood of advanced disease at diagnosis included these factors: incorrect symptom attributions and risk estimations; reluctance to consider the threat posed by the symptom; failure to tell another person about the symptom; and expectations of abandonment by male partners, deportation, prejudice, and refusal of treatment due to poverty. Stories of advanced breast cancer also told of reliance on alternative healing, concerns about overwhelming family resources, and extreme modesty that inhibited obtaining a physical examination. Interventions aimed at earlier detection of breast cancer must connect with the beliefs and assumptions embedded in these narratives, provide pragmatic solutions for perceived constraints on seeking evaluations of self-discovered symptoms, and explore the use of community narratives to confirm the value of early detection of breast cancer. PMID- 9849002 TI - Exploring the barriers to cervical screening in an urban Canadian setting. AB - Cervical cancer is potentially one of the most preventable cancers. The benefits of cervical screening come to those who are actually screened. Despite the known value of cervical screening, a significant number of women do not avail themselves of the procedure. This study examined the barriers to cervical screening in an urban Canadian setting. Focus group methodology was used to explore the perspectives of socioeconomically disadvantaged women regarding their access to health care. The sessions were tape-recorded, and subsequent content analysis revealed four broad themes: being able to talk with doctors is important; being treated as a person is important; finding answers to many questions about cancer is important; and having a Pap test is uncomfortable. Implications for practice and program design were highlighted. PMID- 9849003 TI - Anorectal malformations--update 1998. AB - Anorectal malformation is a common congenital anomaly. Its correction still challenges the wisdom and expertise of the pediatric surgeon. From 1977 through 1997, approximately 400 cases of this anomaly were treated at this institution. Prior to 1985, cut-back anoplasty and translocation anoplasty were the treatments of choice for the low anomalies, and abdominoperineal pull-through anoplasty or sacroperineal anoplasty (and their modifications) preferred for the intermediate or high defects. Since 1985, posterior sagittal anorectoplasty (PSARP) has been adopted exclusively as the operative procedure for the correction of the entire spectrum of anorectal malformations. Two hundred patients have undergone this procedure either primarily (N = 166) or secondarily (N = 34). PSARP is a procedure which enables the surgeon to repair the pelvic anatomy under direct vision, maximally utilizing all the existing continence muscles and hence resulting in better fecal continence. With minimal, limited or maximal division of the continence muscles under one same skin incision, the entire spectrum of this defect can be repaired. The result of cosmesis is generally regarded as better, and the immediate postoperative complications are fewer. In spite of all efforts, 10 to 30% of patients still suffer from total fecal incontinence which is intractable to all kinds of medications and enemas. A bowel management program with large volume enemas, either through the anus or a Malone appendicostomy, retrogradely or antegradely, to wash out stool in the colon and rectum may be necessary to keep the patient clean and more socially acceptable. Biofeedback therapy offers another alternative to improve continence with the development of better computer systems and manometry equipment. This review provides a new concept of the fecal continence mechanism and an operative procedure for constructing a new anorectum based on this concept for the treatment of patients with anorectal malformations. PMID- 9849004 TI - Detection of diabetic retinopathy using non-mydriatic fundus photography at Chang Gung Memorial Hospital. AB - BACKGROUND: Despite recent improvements in the care of diabetic patients, diabetic retinopathy remains the most common cause of blindness among the diabetic population. The aim of this study is to identify the incidence and severity of diabetic retinopathy among diabetic patients followed-up at the metabolic clinics of the Chang Gung Memorial Hospital in Taoyuan, Taiwan. MATERIALS AND METHODS: We investigated the retinas of diabetic patients who were followed-up at the metabolic clinics and received non-mydriatic fundus photograph from April 1994 through June 1994. RESULTS: There were 694 patients with type 2 diabetes enrolled in this study. Their mean age was 56.2 +/- 11.1 years and the mean number of years since diagnosis was 5.9 +/- 5.7 years. Among them, 171 (25%) patients had diabetic retinopathy, including 109 (15.7%) background, 45 (6.5%) preproliferative and 17 (2.4%) proliferative cases. The presence of diabetic retinopathy correlated with the number of years since diagnosis of these diabetic patients (odds ratio: 1.03; p = 0.0024). CONCLUSION: Non-mydriatic fundus photography is a good screening method for diabetic retinopathy detection. Twenty five percent of type 2 diabetic patients followed-up at our metabolic clinics had diabetic retinopathy with background retinopathy being the most predominant. The most significant risk factor of diabetic retinopathy is the number of years since the patient was diagnosed with diabetes. PMID- 9849005 TI - Serodiagnosis of tuberculosis by enzyme-linked immunosorbent assay for anti-A60 and anti-A38. AB - BACKGROUND: For early diagnosis of tuberculosis (TB), especially in the patients without adequate sputum specimens for examination, we found a simple, rapid and inexpensive method among many current available diagnostic tools, the enzyme linked immunosorbent assay (ELISA). To investigate the diagnostic effectiveness of this method, we applied ELISA for detection of antigen 60 IgG and IgM as well as antigen 38 IgG antibodies at Chang Gung Memorial Hospital from April 1995 through June 1996. MATERIALS AND METHODS: Sixty-seven patients were enrolled and divided into 3 groups, Group A (n = 24), patients with positive sputum acid-fast stain; Group B (n = 18), patients with lung cancer and negative sputum acid-fast stain; and Group C (n = 25), patients with chest roentgenogram (CXR) which were suggestive of TB but with negative acid-fast stain results or no sputum for examination. RESULTS: For the A60 IgG antibody, we found a sensitivity rate of 91.7% for Group A and Group B, and 85.7% for Group C as well as an overall sensitivity of 89.5% but with lower specificity. For the A60 IgM antibody, a lower sensitivity (37.5%, 14.3%, 28.9%, respectively) was found but with higher specificity. For the A38 IgG antibody, we found a lower sensitivity (40%, 11.1%, 31%, respectively) but with higher specificity (100%, 71.4%, 90%, respectively). CONCLUSION: With a high sensitivity but low specificity for diagnosis of TB, A60 IgG ELISA could be used as a rapid, simple screening test for patients with results suggestive of TB, especially in those who had no sputum or had negative sputum acid-fast stain results. Otherwise, A60 IgM or A38 IgG ELISA, with a high specificity, could be used as a reliable test in the diagnosis of pulmonary TB when the result is positive. In summary, although ELISA is a simple, rapid, inexpensive method, it is helpful but limited in the diagnosis of pulmonary TB. PMID- 9849006 TI - Effect of postprepared sperm parameters and insemination specimen volume on the outcome of intrauterine insemination. AB - BACKGROUND: The purpose of the present study was to identify the postprepared sperm parameters affecting the outcome of intrauterine insemination and to find out whether the volume of insemination specimen was a determinant factor in the rate of successful conception. MATERIALS AND METHODS: A retrospective study including 306 couples was designed. The patients were inseminated with either 1.0 or 0.5 ml of prepared specimens. The pregnancy rates were compared using the chi square test. Logistic regression was chosen for multivariant analysis of the parameters. RESULTS: The only parameter significantly affecting the success rate was the postprepared sperm motility (p = 0.033). The pregnancy rate was 27.91% in cases with > or = 95% sperm motility. Only two patients with less than 75% sperm motility conceived. The pregnancy rates in cases with 0.5 ml and 1.0 ml inseminations were 12.12% and 16.13%, respectively. This difference was statistically insignificant (p = 0.427). CONCLUSION: The postprepared sperm motility was the only parameter predicting the successful rate of intrauterine insemination. Seventy-five percent sperm motility can be used as a cut-off value for selecting patients. The volume of insemination specimen did not influence the outcome. Insemination with 1 ml of fluid was just as effective as insemination with 0.5 ml. PMID- 9849007 TI - Effects of insulin therapy on non-insulin-dependent diabetics with secondary oral hypoglycemic agent failure. AB - BACKGROUND: To search for the mechanisms of secondary oral hypoglycemic agent (OHA) failure in non-insulin-dependent diabetic (NIDD) and to identify the effects of insulin therapy on these patients, we observed their pancreatic beta cell function and metabolic parameters. MATERIALS AND METHODS: The clinical characteristics and C-peptide responses to glucagon were analyzed in 37 NIDD with secondary OHA failure. All patients received insulin therapy and we measured their fasting plasma glucose (FPG), body mass index (BMI) and glycohemoglobin (HbA1C) levels at 3 months, their serum triglyceride and cholesterol levels at 6 12 months and performed glucagon tests at 0.5-29 months. RESULTS: Poor beta-cell function was observed in 11 (29.7%) patients (Group A) and fair beta-cell function in 26 (70.3%) patients (Group B). The mean age at diagnosis, the number of years since diagnosis of diabetes, FPG levels and HbA1C levels were similar in both groups. However, the patients in Group A had a lower average BMI (18.9 +/- 0.7 vs 22.1 +/- 0.6 kg/m2, p = 0.006) than those of Group B. After insulin therapy, the FPG levels (215 +/- 14 vs 282 +/- 12 mg/dL, p < 0.001) and HbA1C levels 9.2 +/- 0.5 vs 11.6 +/- 0.5%, p < 0.001) of both groups decreased but the BMI (23.1 +/- 0.7 vs 21.6 +/- 0.7 kg/m2, p < 0.001) increased significantly. However, the blood lipid levels and beta-cell function did not change. The beta cell function was still poor in 5 out of 6 patients in Group A, but it was unchanged in all patients in Group B. CONCLUSION: About 1/3 of NIDD with secondary OHA failure are permanently insulin deficient and need long-term insulin treatment. PMID- 9849008 TI - Midtrimester maternal serum free beta-human chorionic gonadotropin levels: normal reference values for Taiwanese women. AB - BACKGROUND: The purpose of this study was to establish normative median values for maternal serum free beta-human chorionic gonadotropin (beta-hCG) during the second trimester of pregnant Taiwanese women. MATERIALS AND METHODS: We collected the data of midtrimester serum free beta-hCG concentration levels from 15,132 normal Taiwanese singleton pregnancies between 14 and 22 weeks' gestation. The maternal age on the day of delivery and maternal weight at the time of sampling were recorded in all cases. The relationship between gestational days and multiple of the median (MoM) levels of serum markers was analyzed using nonlinear regression methods. RESULTS: The median values of free beta-hCG in pregnant Taiwanese women were higher than those of pregnant white women. The log10 standard deviation distribution of free beta-hCG MoM values in the study population was 0.275 with a log10 mean of 0.0097. There were 1.81% of pregnancies with free beta-hCG levels less than 0.3 MoM and 11.33% with levels less than 0.5 MoM; 12.91% had serum levels greater than 2.0 MoM and 5.29% had serum levels greater than 3.0 MoM. CONCLUSION: Because of differences in race and methodology, any laboratory intended to provide determination of free beta-hCG for Down syndrome screening should establish its own normal reference values. PMID- 9849009 TI - Effect of sensory neuropeptides on mucus secretion from cultured goblet cells. AB - BACKGROUND: Airway goblet cell secretion is under the control of capsaicin sensitive sensory nerves. Administration of capsaicin or antidromic stimulation of nerves can stimulate goblet cell secretion and also induce plasma exudation and smooth muscle contraction via a release of neuropeptides from sensory nerve endings in the airway. This study was designed to explore whether the effect of capsaicin or neuropeptides on goblet cell secretion is direct or secondary to other airway responses. MATERIALS AND METHODS: We examined the effects of sensory neuropeptides including substance P (SP), neurokinin A (NKA) and neurokinin B (NKB), as well as capsaicin on cultured guinea pig tracheal goblet cells by measuring the rate of mucus discharge visually under a microscope and the release of 35SO4 radiolabeled mucin-like glycoconjugates. RESULTS: Goblet cells in culture spontaneously secreted intracellular granules at a mean rate of about 3 times/min. SP and NKA increased secretion in a dose-dependent manner. SP was more potent than NKA with maximum responses of 52% and 37% at 10(-8) M, respectively. Quantitative measurements of 35SO4 radiolabeled mucin-like glycoprotein (MLGP) secreted by goblet cells also revealed an increase (109.2%) in mucin secretion caused by SP (10(-8) M) compared with the vehicle control. Neither capsaicin nor NKB caused any significant change in the goblet cell secretory rate. The effects of SP on the secretory rate or release of radiolabeled mucin were not potentiated by an enkephalinase inhibitor, thiorphan (10(-5) M). CONCLUSION: Sensory neuropeptides SP and NKA directly stimulate goblet cell secretion, probably through the tachykinin receptor of the NK-1 subtype as suggested by a greater potency of SP. There was no direct effect of capsaicin on goblet cell secretion. PMID- 9849010 TI - Implications of a failed prospective trial of adjuvant therapy after radical hysterectomy for stage Ib-IIa cervical carcinoma with pelvic node metastases. AB - BACKGROUND: Lymph node metastasis is established as a poor prognostic factor in cervical carcinoma patients undergoing primary surgery. However the optimal postoperative therapy for node-positive patients remains to be defined. MATERIALS AND METHODS: To determine the role of adjuvant therapy in stage Ib-IIa cervical carcinoma patients who had pelvic node metastases after radical hysterectomy, a single institutional randomized controlled trial with factorial design, comparing concurrent chemo-radiotherapy (CT + RT) versus chemotherapy (CT) or radiotherapy (RT) alone was conducted. Study endpoints included site of recurrence, time to recurrence, relapse-free and overall survivals, and toxicity of treatment. RESULTS: At a median follow-up of 61 months, the 6-year relapse-free and overall survival rates of the 39 patients entered into this trial were 78.2% and 85.2%, respectively. This trial was prematurely closed due to suboptimal accrual and protocol violation. It is inconclusive regarding the efficacy of CT because of the limitation of the sample size and the imbalance of prognostic features by actual treatment. A model for risk group classification of patients with stage Ib II cervical carcinoma with pelvic node metastases from a retrospective analysis was validated by this prospective cohort. The results of this failed trial suggest that adjuvant CT alone seemed comparable to RT alone or CT + RT in survival but was associated with significantly less morbidity. CONCLUSION: It is warranted to consider a CT alone arm in comparison with either observation or CT + RT according to risk of recurrence in future prospective trials. However, this important issue can only be addressed by a large multicenter trial. PMID- 9849011 TI - Cementless ceramic total hip arthroplasty: a 5 to 16 year follow-up. AB - BACKGROUND: Cementless total hip arthroplasty (THA) was invented to replace cemented THA and thus avoided cement disease, which shortened the longevity of implants. Ceramic THA was an early type of cementless THA which was used at our institution for several years. In the literature, reports about ceramic THA are very rare, moreover, the results of THA are controversial. This retrospective study was to report our experience. MATERIALS AND METHODS: Seventy-two cementless ceramic THAs were performed and followed for at least 5 years (range, 5 to 16 years; average, 9.8 years) at our institution. Patients ranged in age from 23 to 68 years (average, 45 years) with a male to female ratio of 3 to 1. A modified d'Aubigne and Postels' method was used to evaluate the functional and radiographic outcomes. RESULTS: Only 51.4% of the cases achieved a satisfactory result. Furthermore, the complication rate was very high, especially for aseptic loosening (34.7%). For ankylosing spondylitis cases, the satisfactory result rate was only 30% (2/7) and the unsatisfactory cases even had a loosening rate of 80% (4/5). CONCLUSION: Compared to other types of cementless THA, the satisfactory result rate with ceramic THA was markedly lower and the complication rate was relatively higher. Therefore, we conclude that cementless ceramic THA is not an ideal implant and its usage should be strictly restricted. PMID- 9849012 TI - Adenosine triphosphate modification of endotoxin-induced otitis media with effusion in gerbils. AB - BACKGROUND: Impairment of the mucociliary system of the middle ear epithelium is a key step in developing otitis media with effusion. Effective propulsion of the viscoelastic gel layer of mucus requires a control of the depth of the periciliary sol layer by means of epithelial ion transport. In a previous study, we successfully demonstrated modulation of ion transport via short-circuit current changes by specific stimulation of adenosine triphosphate (ATP) receptors in a middle ear cell line. MATERIALS AND METHODS: Otitis media with effusion was developed in gerbils by infusing Escherichia coli E.coli lipopolysaccaride. In group A, E. coli lipopolysaccharide was first infused, and after establishment of middle ear fluid, the right ear was infused with 10(-4) M ATP, while the left ear remained as the control. In group B, the right ear was infused first with 10(-4) M ATP followed by bilateral infusion with endotoxin. The morphology of the middle ear mucosa was studied using scanning electron microscopy (SEM). RESULTS: Group A showed fluid clearance in 5 out of 15 ATP-treated right ears while 3 out of 15 left ears showed clearance. Group B showed persistent presence of fluid in 11 out of 15 ATP-treated right ears, while the left ears showed fluid in 13 out of 15. SEM study consistently demonstrated scanty cilia in the non-ATP treated ears, but in ATP-treated ears the cilia remained abundant. CONCLUSION: Although the results showed that ATP could not effectively prevent or clear middle ear effusion, the SEM result showed a marked beneficial effect of ATP on preserving ciliary configuration. PMID- 9849014 TI - Malacoplakia in a colonic adenoma: case report. AB - We report a case of colonic malacoplakia in a tubular adenoma. A 67-year-old neurotic man had been treated with minor tranquilizers for 10 years. In January 1997, he underwent a health examination. Most of the tests performed were within normal limits, except the colonoscopy, which revealed a polypoid lesion in the left colon. A polypectomy was performed at that time. The histopathological study of the polyp showed a tubular adenoma with malacoplakia, characterized by an aggregate of numerous histiocytes containing Michaelis-Gutmann bodies in the lamina propria. A review of the English literature indicated that 35 cases of colonic malacoplakia have been documented. Colonic malacoplakia can occur in isolation or with diffuse or multiple involvement of the adjacent and unrelated diseased sites. The majority of cases are associated with other diseases, the most common being carcinoma of the colorectum. The association of colonic malacoplakia with adenoma is extremely rare and only 3 cases including our present case have been reported. We suggest that colonic malacoplakia is more common than usually suspected. A careful histologic examination is necessary. PMID- 9849013 TI - High-grade endometrial stromal sarcoma in a 10-year-old girl: case report. AB - Endometrial stromal sarcomas occur primarily in perimenopausal women and are quite uncommon in children. We report a case of high-grade endometrial stromal sarcoma in a 10-year-old girl, who initially experienced lower abdominal pain on voiding. After a series of examinations, surgical treatment was performed, including an extended total hysterectomy with bilateral salpingo-oophorectomy, bilateral pelvic lymph node sampling, and partial omentectomy. Adjuvant radiotherapy and hormonal therapy followed, however, the abdominal tumor recurred 9 months later. Invasion of the right lobe of the liver with compression of the inferior vena cava and pleural effusion was noted. The patient then received prolonged oral etoposide therapy. The treatment stabilized the tumor size and relieved her symptoms for 4 months. She finally succumbed to the disease as a result of distant metastasis. PMID- 9849015 TI - Pregnancy in a previous cesarean section scar: case report. AB - An ectopic pregnancy developing in a previous cesarean section scar is an extreme rarity among all ectopic pregnancies. Due to the difficulty of making an accurate diagnosis, this kind of ectopic pregnancy is the most dangerous and life threatening type. A 30-year-old woman, who was gravida 4, para 3, was admitted to our emergency room with massive vaginal bleeding. She had undergone a cesarean section due to a breech birth 3 months prior to this admission. Ultrasound examination showed a pregnancy located in the low corpus uterus. The possibility of a spontaneous abortion in progress or a cervico-isthmic pregnancy were considered. Due to the massive vaginal bleeding and unstable vital signs of the patient, surgical intervention was decided upon to save the woman's life. We first performed dilatation and curettage. There was no gestational villi and severe vaginal bleeding was noted during this procedure. An emergent exploratory laparotomy was then performed, followed finally by a hysterectomy. Pathologic findings confirmed the diagnosis of a pregnancy in the previous cesarean section scar. Diagnosis, prevention, clinical evaluation and management of these conditions are discussed. PMID- 9849016 TI - Clinical courses and changes of pancreatic beta-cell function in young-onset diabetics: report of two cases. AB - Accurate classification of diabetes in some young patients is difficult but clearly of importance in deciding the appropriate treatment. We report the different clinical courses and beta-cell function changes in 2 young-onset diabetics. In the beginning, each of them was considered to be a patient with insulin-dependent diabetes mellitus (IDDM) and was treated with insulin. Then, treatment was shifted from insulin to oral hypoglycemic agents (OHA) because each patient's beta-cell function was comparable to that of patients with non-insulin dependent diabetes mellitus (NIDDM). In case 1, the patient's glycemic control and beta-cell function progressively deteriorated and finally insulin therapy was required. In case 2, the patient was continuously treated with OHA. Although his blood glucose was not well-controlled, his beta-cell function had been well preserved. Recently he achieved near-normoglycemia by improving his diet and OHA compliance. Clearly, case 1 was a slowly progressive Type 1 DM patient and case 2 was a young Type 2 DM patient. This classification was further supported by the finding that the patient in case 1 had human leukocyte antigen (HLA)-DR3, but the patient in case 2 did not. PMID- 9849017 TI - Airway compression by a biplane pediatric transesophageal echocardiography probe: case report. AB - Transesophageal echocardiography (TEE) has been extensively used in cardiac surgery to assess cardiac function and anatomical relationships in both adults and children. Newer monitoring devices, despite being labeled "noninvasive", often present significant risks. Physicians should be cautious with the use of TEE in infants and small children, recognizing that oversized probes may inadvertently compress vital structures, resulting in airway or vascular compromise. If severe compression of the airway or vascular structures occurs, TEE monitoring may need to be discontinued. Although technological progress has yielded smaller ultrasonic probes, the TEE probe is significantly larger than most other devices placed in the esophagus. We present in this report a case of airway compression using a small-sized biplane pediatric TEE probe in a child undergoing Blalock-Taussig shunt surgery. Pediatric patients may be more at risk for airway obstruction because of the over-size of the probe-distended esophagus in relation to adjacent airway structures. PMID- 9849018 TI - Giant-cell tumor of the patella: report of two cases. AB - Two patients with giant-cell tumors of the patella are presented in this report. Both patients were young females who were noted to have had nonspecific anterior knee pain and mild swelling of 1 to 12 months' duration prior to admission to our hospital. Local tenderness over the peripatellar area and slight limitation of full flexion were noted during physical examination. The radiographic presentation of each patella appeared as an expansile and lytic lesion with a thin cortex, without evidence of intra-articular involvement. Chest radiography and routine laboratory examination results were normal. After biopsy, intralesional curettage with phenol cauterization and allograft reconstruction was the preferred treatment in these two patients, with both tumors considered to be stage 2 according to Enneking's staging system. Following surgery, range of motion exercise was started after 6 weeks of immobilization with a long leg splint. Both patients regained full range of motion and were pain free. Radiographically, bone remodeling without evidence of recurrence was noted in both patients 2 years postoperatively. PMID- 9849019 TI - Traumatic thoracic aortic injury caused by a sharp edge of left fractured rib on body position change: case report. AB - Aortic injury caused by penetration of a fractured rib is very rare. We present a patient with aortic injury demonstrated using serial imaging studies. A 66-year old woman fell from a ladder and sustained multiple left-side rib fractures. There was a small left hemothorax and widened mediastinum on the initial chest roentgenogram in the emergency department. Chest computed tomography (CT) revealed a posterior segmental fracture of the sixth rib on the left side with a sharp edge penetrating into the posterior aspect of the thoracic aorta. It was initially missed. More than 1000 cc of fresh blood suddenly gushed out of the chest tube 7 hours after the traumatic event. After resuscitation, an aortogram was performed which showed blood extravasation from the thoracic aorta at the rib fracture site. Unfortunately, surgical intervention was delayed and she died. Early detection and early surgical intervention are necessary in patients with a widened mediastinum and positive results on imaging studies. PMID- 9849020 TI - Multiple lymphomatous polyposis of the gut: case report. AB - Multiple lymphomatous polyposis (MLP) is an uncommon type of primary non Hodgkin's gastrointestinal B-cell lymphoma characterized by the presence of multiple lymphomatous polyps along the gut. We present a patient with MLP in which the involvement was unusually widespread. The diagnosis was confirmed by the typical polyposis lesion, histology, phenotyping and clinical presentations. A 68-year-old man had a large mass at the ileocecal valve as well as multiple polyps along the whole digestive tract. At the time of diagnosis, lymphoma had involved bone marrow, peripheral blood, spleen, prostate and peripheral lymph nodes. The patient received 8 courses of chemotherapy with no remission. He died of pneumonia 11 months after diagnosis. Clinically, the diagnosis may be confused with epithelial polyps; and histologically, the diagnosis must be distinguished from benign lymphoid proliferations as well as other types of lymphoma. The prognosis for patients with MLP is relatively poor (the median survival is usually less than 3 years). PMID- 9849021 TI - High-grade mucoepidermoid carcinoma of the breast: case report. AB - Mucoepidermoid carcinoma of the breast is very rare. Only 14 cases have been reported in the English language literature. All of these patients were women with ages ranging from 46 to 72 years. Five of the cases involved high-grade mucoepidermoid carcinomas and 9 were low-grade. All of the patients with low grade mucoepidermoid carcinoma were disease-free in the follow-up period. Four patients with high-grade mucoepidermoid carcinoma died from the disease 7 to 30 months after surgery and all 4 were found to have distant metastasis. The case of mucoepidermoid carcinoma of the breast discussed here was histopathologically high-grade. The patient has remained disease-free 4 years after modified radical mastectomy and adjuvant chemotherapy. Diagnose of mucoepidermoid carcinoma of the breast is more difficult than mucoepidermoid carcinoma in the salivary gland due to possible confusion with metaplastic carcinoma or squamous cell carcinoma. Special histological stains are definitely needed for differential diagnosis. PMID- 9849022 TI - Inferior dislocation of the glenohumeral joint combined with the compartment syndrome of the upper arm: case report. AB - Compartment syndromes occurring in the forearm and leg are not infrequent. However, reports of compartment syndrome of the upper arm are conspicuously rare. Inferior dislocation of the shoulder combined with compartment syndrome has rarely been reported in the literature. We report our experience with a patient with inferior dislocation of the glenohumeral joint combined with compartment syndrome of the upper arm. A 29-year-old man had traumatic inferior dislocation of the glenohumeral joint combined with severe swelling of the shoulder and upper arm. After close reduction, the shoulder and upper arm were still tensely swollen. Compartment pressure of the upper arm was measured using a digital manometer (Quick Pressure Monitor 295-1, Stryker) and remained very high, thus, fasciotomy was performed. After debridement and skin graft, the arm healed without sequelae. When a patient has a fracture dislocation of the shoulder joint associated with a swollen arm, compartment syndrome of the upper arm should be included as a differential diagnosis. Detecting the compartment pressure can confirm the diagnosis of compartment syndrome objectively. PMID- 9849023 TI - Penicillium marneffei fungemia in an AIDS patient: the first case report in Taiwan. PMID- 9849024 TI - Aetiology of nasopharyngeal carcinoma. A review. AB - The present overview is based on recent available information on nasopharyngeal carcinoma (NPC) from various parts of the world including India. The available data suggests that NPC is a rare tumour in most parts of the world. But the incidence is higher in China and South East Asia and also among the Chinese wherever they have migrated. NPC is also relatively higher especially among the mongoloid group of the people in the North Eastern Region of India as compared to other parts of the country. The distinct geographical and ethnic distribution of NPC have stimulated much research to find out its etiology. The results suggest that Epstein Barr Virus (EBV) infection and genetic susceptibility are the constant aetiological factors for the higher incidence of NPC among various ethnic groups while other factors such as ingestants and inhalants may depend on the distinct dietary practices and living environment adopted by various ethnic groups in different geographical region of the world. PMID- 9849025 TI - Descriptive epidemiology of thyroid cancer in greater Bombay. AB - In this paper an attempt has been made to present a descriptive epidemiology of thyroid cancer in Bombay, and it is discussed in relation to age, demographic and socio economic composition of the population, using the most recent five year date. Time trend analysis of this cancer by sex has also been discussed using the last 30 years data. When international incidence of thyroid cancer was ranked in descending order for various countries, the incidence recorded for Bombay was found to be at the lowest level in both the sexes. Thyroid cancer is about three times more frequent among women than men, but this relative excess varies with the histologic type and age. As in the case of the majority of cancers, the incidence curve for thyroid cancer rises with age. However in men, the increase continues consistently with advancing age, while in women it begins to level off after the age 30, leading to an almost equal sex ratio in old age. In Bombay the incidence of thyroid cancer in men was found to be the highest in Muslims and in Christian women. No association was observed between thyroid cancer and education level attained by these patients. The four main histologic types of thyroid cancers i.e. papillary, follicular, anaplastic and medullary are also observed in Bombay. It has been noted that there is an increasing trend in the age-adjusted incidence rate for thyroid cancer in both the sexes in Bombay in the period under review 1964 to 1993. But the increase in incidence was found to be statistically significant only in males. PMID- 9849026 TI - Risk assessment of tobacco, alcohol and diet in cancers of base tongue and oral tongue--a case control study. AB - This is a retrospective case-control study of male tongue cancer patients seen at Tata memorial Hospital, Bombay, during the years 1980-84. The purpose of the study was to identify the association of tobacco, alcohol, diet and literacy status with respect to cancers of two sub sites of tongue namely anterior portion of the tongue (AT) (ICD 1411-1414) and base of the tongue (BT) (ICD 1410). There were 142 male AT patients and 495 BT patients interviewed during the period. 635 interviewed male patients who were free of any disease were considered as control. Bidi smoking was found to be a significant risk factor for BT patients and tobacco chewing for AT patients respectively. Alcohol drinkers showed about 45% to 79% excess risk for both sites of tongue cancer. Illiteracy and non vegetarian diet proved to be a significant factor for AT patients only. The study brings out that the location of cancer has got a direct bearing with the type of tobacco use and other related habits and this inturn may provide meaningful interpretation of variations observed in the incidence of tongue cancer around the world. PMID- 9849027 TI - Ectopic ACTH syndrome due to bronchial carcinoid tumour--a case report. AB - A patient is reported who had Cushing's syndrome and carcinoid tumour of the bronchus. The case illustrates the difficulty in preoperative localization of the ectopic ACTH source and the surgical management of such patients. PMID- 9849028 TI - Polymyositis--an unusual presentation of ovarian malignancy: a case report. AB - A unique case of carcinoma of ovary with an unusual presentation is reported here. Her presenting complaint was in no way related to ovarian malignancy. She presented to the neurologist with the features suggestive of polymyositis. She did not respond satisfactorily to the conventional treatment by steroids and on investigation, was found to have an underlying advanced ovarian malignancy. PMID- 9849029 TI - Comparison of magnified chemiluminescent examination with incandescent light examination and colposcopy for detection of cervical neoplasia. AB - Magnified Chemiluminescent Examination (MCE) or speculoscopy is a new visual method for detection of cervical neoplasia. It utilizes low magnification and a special "blue-white" chemiluminescent light. The study includes 125 women with unhealthy cervix who were subjected to magnified examination with projected incandescent light (PIL) and chemiluminescent light (MCE). This was followed by colposcopy and directed biopsy from acetowhite areas. Of all 125 subjects, 20 patients showed cervical neoplasia of varying degree on histopathology. MCE could detect 18/20 neoplasias while PIL detected 11/20 cases. Sensitivity of MCE (90%) was significantly superior (P < 0.05) to PIL (55%) in detecting cervical neoplasia. Colposcopy, as compared to MCE, is better than speculoscopy as it facilitates grading of lesions due to higher magnification but antecedent MCE detected the acetowhitening in majority of patients (32/43). However, tendency to "overall" acetowhite lesions was significantly less during MCE when compared with colposcopy. Thus, MCE is a new diagnostic tool which is better than routine incandescent light examination and correlates well with colposcopy. PMID- 9849030 TI - The submental island flap for reconstruction of facial defects. AB - The procedure of facial resurfacing dictates that there should be an excellent colour and texture match between the facial and the transposed skin. Cervical flaps e.g. platysma flap are commonly used for reconstruction of facial defects but, with disadvantages like limited mobility, unacceptable donor site and unpredictable outcomes. The submental island flap is a new addition to the armamentorium of the Plastic Surgeon. It is an axial pattern flap based on the submental branch of facial artery and the submental tributary of common facial vein. We report the use of this flap for the reconstruction of postexcisional facial defects in two patients having basal cell carcinoma and recurrent parotid tumour. In both patients, the flap provided an excellent colour and texture match. Other advantages of this flap are inconspicuous donor site, wide arc of rotation, simple dissection and a reliable pedicle. PMID- 9849031 TI - Hysteria--a chameleon or a fossil? PMID- 9849032 TI - Chlamydial and gonococcal serology in women with tubal infertility. AB - Sera from 81 infertile women with tubal pathology and 40 controls were tested for the presence of antibodies against Chlamydia trachomatis & Neisserria gonorrhoeae. Indirect immunoperoxidase test (Ipazyme kit) & Enzyme linked immuno sorbent assay (ELISA kit) were used for detection of chlamydial & gonococcal antibodies respectively. Antibodies to Ch. trachomatis were found in 74.07% of the infertile women and 5% in control group. Only a very low prevalence (4.93%) of antibodies to N. gonorrhoeae was found is infertile women as compared to nil in control group. Antibodies detection is a sensitive, specific and noninvasive test for diagnosing infertility. PMID- 9849034 TI - Incidence of HIV risk and prevention of infection in health care settings. AB - Tht HIV epidemic has created awareness about blood borne pathogens transmission in health care settings. Although, the risk of transmission of HIV and other blood borne pathogen may account for a very small proportion, yet the potential for such transmission always exists. Despite this, the use of proper and careful sterile procedures is sufficient to substantially reduce or even eliminate the risk of HIV infection. PMID- 9849033 TI - Double-blind placebo--controlled trial of nandrolone decanoate in postmenopausal osteoporosis. PMID- 9849035 TI - Molybdenum, xanthine oxidase and riboflavin levels in tamoxifen treated postmenopausal women with breast cancer. AB - 40 cases postmenopausal women with breast cancer constituted the study group and 20 sex and age matched formed the control group. The study group of untreated patients showed nonsignificant decrease in molybdenum but significant decrease in blood xanthine oxidase and riboflavin levels. Tamoxifen treated patients showed nonsignificant increase in molybdenum, after 3 months, significant increase after 6 months and significant increase in xanthine oxidase and riboflavin levels. Thus tamoxifen when given in breast cancer helps in amelioration of the diseased condition. PMID- 9849036 TI - Obesity: as rural females perceive it. AB - Out of 318 rural females studied, 17.3% were found to be obese. Most of them are aware of their obese status except for 10.9% who were unaware. Majority of the females considered eating more, childbirth and reduced activity as the possible cause of their obesity and also obesity in other persons. Though 48.1% and 33.6% of the females mentioned dieting and exercise/walking respectively as the method of prevention still 52.5% mentioned that medicine is the only method for prevention of obesity. Regarding remedy of obesity again 48.4% females mentioned medicine as the method. PMID- 9849038 TI - Enzymes--a review. AB - Enzymes are essential to life and catalyze various metabolic activities. They find application in a number of diseased conditions, and their scope is being widened as knowledge develops. PMID- 9849037 TI - Truck crew and risk of contraction of STDS, including HIV: encouraging the use of condoms by the method of applying theories. AB - Studies from India conclude that the long distance truck crew are at a high risk of contracting and transmitting STDs and HIV on account of their high risk sexual behaviours. These studies indicate the necessity of changing the behaviour of the truck crew in favour of safer sexual practices. This article addresses this important issue of developing an intervention to decrease the risk of contraction of STDs, including HIV, by encouraging the use of condoms by the method of applying psycho-social theories, which aim at changing behaviours of the target group. PMID- 9849039 TI - Prognostic analysis in patients of lung cancer with brain metastasis under surgical removal. AB - The outcome and prognostic factor in postcraniotomy patients with brain metastasis from lung cancer in south Taiwan were evaluated. There were 21 males and 12 females, with ages ranging from 35 to 72 years (mean, 56.4 years). The survival rate was determined from the date of craniotomy and was evaluated by the Kaplan-Meier product limit method. In the univariate analysis survival was significantly affected by systemic metastasis (P = 0.0109), brain radiotherapy (P = 0.0121) and chest radiotherapy (P = 0.0361). Preoperative and postoperative Karnofsky scores were evaluated using the paired t-test and the neurological condition, as evaluated by Karnofsky scores, significantly improved after craniotomy (P < 0.001). We concluded that an aggressive craniotomy and radiotherapy improves the quality of life and survival rate, especially in those patients with solitary brain metastasis and no extracranial metastasis. PMID- 9849040 TI - The experiences of rectosigmoid pouch as a continent urinary diversion after radical cystectomy. AB - Thirty-five patients receiving cystectomy underwent rectosigmoid pouch. The technique of ureteral submucous implantation was described. The follow-up results were analyzed. The relationship of postoperative voiding frequency and preoperative anal closure pressure was assessed. The day and night time continence rate is 97% (34/35). There were no hydronephrosis, no ureterocolonic stricture or reflux. The complications are not unique to rectosigmoid pouch and not high when compared with other forms of urinary diversion. When the preoperative anal closure pressure reaches nearly 100 cmH2O a postoperative outcome with good quality of life will be predicted. With the advantages of high continence rate and simplicity of performance the rectosigmoid pouch will become one of the alternative forms of continent urinary diversion. For obtaining the favorable results a patient with normal renal function, good hepatic function and anal closure pressure more than 50 cmH2O is required. PMID- 9849041 TI - Health and satisfaction among the elderly with chronic conditions: demographic differentials. AB - This study examines the state of health (physical and mental health) and avowed satisfaction (toward hospital services and towards life-as-a-whole), among 172 elderly subjects (108 outpatients from a family medicine clinic and 64 community residents) with chronic conditions living in Kaohsiung city. Results indicated that (a) the elderly did not perceive any great impact of illness(es) upon their lives and both their physical and mental health were satisfactory; (b) the elderly were generally satisfied with both the hospital services and life as a whole; (c) there were meaningful group differentials in health and satisfaction: older, male, single and financially-dependent elderly had poorer physical and mental health; female, single and financially dependent elderly avowed lower life satisfaction. PMID- 9849042 TI - Revised Taiwanese Elderly Stressor Inventory. AB - The purpose of this study was to revise the 73-item Taiwanese Elderly Stressor Inventory (TESI) so as to shorten the time needed by professionals and researchers to assess the stress experience of elderly persons. The research included two phases: Phase I consisted of revising the original form by combining similar items and deleting some TESI items which were found not to effectively test the concept of stress; and Phase II analyzed the reliability and validity of the revised form. The revised instrument consists of sixty stressor items. Two doctorally prepared nurses were used to determine TESI item content validity, resulting in a Content Validity Index of 0.97. Three hundred and one subjects from one district in Metropolitan Taipei area were used in determining the psychometric properties of the revised form. A Cronbach alpha of 0.74 was found. The coefficient for test-retest reliability was 0.89. As to construct validity, correlations of -0.327 with health perception, 0.520 with depression and -0.433 with life satisfaction were found. In the multiple regression analysis, health perception, depression, stressfulness and social support were predictors, while life satisfaction was a criterion variable. Total explanation variance was 41.42%. These results reveal the predicted pattern of relationship among variables, which reflected the modest validity of the revised TESI. The revised TESI possesses a sound psychometric properties, which suggests that it can be readily used in practice settings. PMID- 9849043 TI - Pre-analytical for biological monitoring of manganese. AB - Manganese is an essential element but it may result in a toxic event if the workers are excessively exposed to this element. Monitoring of biological Mn exposure is therefore important. The careful preparation for Mn analysis is necessary. In order to get a correct protocol for sampling and storage of the biological samples, identification of the possible contamination source is necessary. This study is concerned with the monitoring procedure of manganese especially the sources of contamination sampling, sample transportation and storage. The results of the study shows that some kinds of needles and anticoagulants might be the main sources of contamination of specimen. It is important to avoid these contamination in performing biological monitoring on workers exposed to manganese. PMID- 9849044 TI - Craniofacial pattern of young adults with various types of malocclusion. AB - Craniofacial morphological characteristics of young adults with various malocclusions were investigated. Cephalometric radiographs of 100 Class I, 100 Class II, and 100 Class III cases were selected and subjected to counterpart analysis as described by Enlow. Results of individuals with normal occlusion revealed a mean value of 38.75 degrees for the angle of the middle cranial fossa relative to the posterior maxillary plane alignment (MCF/PM). This value was used as the basis for intrinsic alignment comparisons for all the other various parts. Classes I and II malocclusions were classified as either type A or B, depending on whether point A or B was protrusive in the functional occlusal plane (FOP). We found that type B was more common for Class I malocclusion. Further, these cases exhibited an underlying Class III character. We also noted that B groups had different structural craniofacial patterns relative to those seen in A groups. The difference between Class IA and Class IIA, or between Class IB and Class IIB was slight but quantitative. Class III and Class IIA individuals had distinctly different and essentially opposite underlying patterns. However, the composite compensatory result in Class IB case was, for the most part, effective. PMID- 9849045 TI - Diastolic mitral regurgitation secondary to aortic regurgitation--a case report with emphasis on its mechanism. AB - We report on a 68-year-old male case of mitral regurgitation secondary to aortic regurgitation diagnosed using transthoracic echocardiography. A different mechanism of mitral regurgitation secondary to aortic regurgitation is demonstrated in this report. Two-dimensional color Doppler echocardiography may be a useful tool for anatomic evaluation to define its mechanism. PMID- 9849046 TI - Trigeminal neurinoma presenting as intratumoral hemorrhage--case report. AB - Neurinomas of trigeminal nerve are rare intracranial tumors and spontaneous intratumoral hemorrhage occurred exceedingly rarely in them. Herein, we report one case of trigeminal neurinoma presenting spontaneous intratumoral hemorrhage. Presenting symptoms were headache, lethargy and vomiting of sudden onset. Unenhanced computed tomography scans showed a high density mass in the left cerebellopontine angle cistern. On magnetic resonance images, they revealed hyperintensity on both T1- and T2-weighted images. A left retromastoid suboccipital craniectomy was performed and nearly total tumor removal was achieved. Histologically, increased vascularity, dilated vessels and hemosiderin were observed in the specimen. Tumor size, hypervascularity and vascular change are important factors for hemorrhage. PMID- 9849047 TI - Bone regeneration in advanced periodontitis lesions after a single episode of root planing--case report with ten-years of follow-up. AB - This report describes a significant bone regeneration in a mandibular molar with advancing long-standing periodontal disease (defect extending to apex and into furcation) following a single episode of scaling and root planing. A 52-year-old female suffered from generalized moderate periodontitis for two years. An acute periodontal abscess with a 12 mm probing depth around the right mandibular first molar was noted. After incision and drainage, thorough scaling and root planing was performed under local anaesthesia. Three months later, radiographic examination showed complete bone regeneration with formation of a distinct lamina dura. It was concluded that favorable clinical and radiographic results could be obtained after a single episode of thorough subgingival root planing combined with excellent oral hygiene maintenance. PMID- 9849048 TI - [Effect of intracoronary dipyridamole administration on the incidence of restenosis after PTCA. A prospective randomized study]. AB - BACKGROUND: Restenosis after PTCA remains a serious long-term complication of balloon angioplasty occurring in 30 to 50% of patients. Platelets play a crucial role in the pathogenesis of restenosis following PTCA. Dipyridamole has been shown to inhibit platelet aggregation in humans. Its action as an antithrombotic drug can be attributed to different mechanisms including inhibition of platelet phosphodiesterase and inhibition of the cellular uptake of adenosine. PATIENTS AND METHODS: The purpose of the following study was to investigate the effect of an intracoronary infusion of dipyridamole on the incidence of angiographic and clinical restenosis. In 763 balloon angioplasties patients were randomly allocated to receive either conventional pretreatment (heparin 15000 IE, aspirin 500 mg i.v.) or an additional intracoronary infusion of dipyridamole (0.5 mg/kg body weight). Conventional pretreatment was performed in 388 interventions (61 interventions in women, age 60.5 +/- 8.7 years; 47 interventions for acute coronary syndromes); in 375 interventions additional intracoronary dipyridamole was infused (58 interventions in women, age = 59.6 +/- 9.6 years; 57 interventions for acute coronary syndromes). RESULTS: As compared to conventional pretreatment intracoronary dipyridamole application was associated with a reduction in angiographic restenosis from 43.0% to 36.8% and a reduction of target vessel revascularisation by 15.5% but failed to reach statistical significance. These results were due to an increase in net gain following dipyridamole application. CONCLUSION: Intracoronary pretreatment with dipyridamole prior to PTCA fails to reduce the incidence of angiographic restenosis and target vessel revascularisation significantly. However, a moderate improvement of long-term follow-up can be achieved. PMID- 9849049 TI - [Reduced incidence of side-effects of growth hormone substitution in 404 patients with hypophyseal insufficiency. Results of a multicenter indications study]. AB - BACKGROUND: Substitution of pituitary insufficient patients with recombinant human growth hormone (rhGH) in addition to the conventional substitution with glucocorticoids, L-thyroxine and sex hormones has been approved by the regulatory authorities in 1995 with the imposition to conduct surveillance studies to monitor drug safety. RESULTS: 24% of all patients were within their 2nd treatment year, 15% within their 4th year, maximum treatment period was 6 years. There were 2 peaks within the patients age distribution: 30 to 39 years (24%) and 50 to 59 years (24%). The causes for pituitary disease were as follows: pituitary adenomas (47%), idiopathic (16%), craniopharyngeomas (16%) and others (21%). Mean GH dose was 1.5 IU/d s.c. (range 0.4 to 4 IU/d). Serum-IGF-1 increased by 159 and 192% in females and males. Waist circumference decreased by 2% and serum cholesterol was lowered by 5.5% in males. There were 2 cases with new carcinomas, 1 diabetes mellitus II and 1 death. Adverse events (AEs) within KIMS were compared to those of the treatment (GH) and placebo (PI) groups of the previous admission trials (in percent): edema: KIMS 10, GH 37, Pl 3; arthralgia: KIMS 8, GH 19, Pl 2; muscle pain: KIMS 3, GH 16, Pl 3; dizziness: KIMS 2, GH 1, Pl 3; headache: KIMS 2, GH 3, Pl 2; others: KIMS 2, GH 22, Pl 13. The reported incidence of AEs in KIMS was lower than in previous clinical trials. There might be 3 reasons for this: 1. under-reporting, particularly those AEs not likely to be related to GH treatment; 2. doses used in trials were 2-fold higher than in KIMS; 3. dose titration for individual patients. CONCLUSION: Surveillance programs are important for monitoring of drug long-term efficacy and safety. PMID- 9849050 TI - [Regulation of T-cell activation by CD28 and CTLA-4]. AB - T CELL RESPONSE: T lymphocytes play a key role in the coordination of the immune response. T helper cells contribute primarily by means of cytokine release, whereas cytotoxic T cells eliminate cells bearing antigens recognized as foreign. Through its T cell receptor each T cell can recognize a specific peptide antigen, which is presented in the context of the major histocompatibility complex (MHC) to T helper cells by specialized antigen-presenting cells or to cytotoxic T cells by nearly all body cells. Upon contact with its specific antigen, the T cell receptor transduces an activation signal into the T cell, leading to proliferation, cytokine production, or efficient cytotoxicity. COSTIMULATION: However, a second costimulatory signal is necessary to achieve complete activation. This can be provided by the accessory T cell molecule CD28 upon binding to its respective ligands B7-1 (CD80) or B7-2 (CD86). The same ligands bind to CTLA-4 (CD152), a receptor expressed at the surface of T cells previously activated for 2 to 3 days and capable of downregulating activation. IMMUNOSUPPRESSION BY CTLA-4Ig: A genetically engineered soluble fusion protein containing the extracellular domain of CTLA-4 and the Fc portion of an immunoglobulin heavy chain (CTLA-4Ig) prevents the interaction of CD28 and CTLA-4 with their B7 ligands, the subsequent activation of T cells and thereby eliminates or reduces unfavorable immune system activation in transplant rejection or autoimmunity. CONCLUSION: The importance of the regulatory system comprising CD28, CTLA-4 and the B7 molecules and its modulation by CTLA-4Ig has been demonstrated in a substantial number of animal models in recent years and holds promise as a novel approach for therapeutic immunomodulation in humans. PMID- 9849051 TI - [Febrile neutropenia: practical aspects]. AB - BACKGROUND: Infections are a major cause of mortality in neutropenic patients. They require long hospital stays and highly expensive therapeutic measures. In this review we discuss the practical and pharmaco-economic aspects of the management of febrile neutropenia. PREVENTION AND THERAPY: Prevention of fever of unknown origin (FUO) demands hygienic and antimicrobiotic measures. First-line antibiotic therapy consists of an aminoglycoside combined with an ureidopenicillin or a 3rd-generation cephalosporin. Double beta-lactam antibiotic combinations are equally effective and less toxic, but more expensive. Monotherapy with carbapenems, ceftazidime, or cefepime appear to offer comparable efficacy. Lung infiltrates require immediate treatment with amphotericin B. If the initial therapeutic regime fails, a carbapenem plus a glycopeptide antibiotic and a parenteral antimycotic drug should be applied after 3 to 4 days. The prophylactic or interventional administration of hematopoietic growth factors is only indicated in special high-risk situations. CONCLUSIONS: Using the described therapeutic procedure, the response rate exceeds 90%. Consistent, step-wise escalating administration of antibiotics is essential. More evaluation is needed to determine whether selected patients with febrile neutropenia can be treated on an outpatient basis. PMID- 9849052 TI - [Spontaneous bacterial peritonitis]. AB - BACKGROUND: Since the beginning of the eighties systematic investigations broadened our knowledge about the clinical picture of spontaneous bacterial peritonitis very much. Important insights into epidemiology, pathogenesis, symptomatology, diagnosis and therapy of this disease, which is a frequent complication in patients with cirrhosis of the liver and ascites, could be gained. Actual research work primarily deals with questions of therapy and prophylaxis. AIM: Aim of this review is a comprehensive presentation of the different aspects of this disease on the basis of the present literature. CONCLUSIONS: As on the one side the clinical symptoms may be very little and on the other side the prognosis is very bad, it is extremely important to take this entity into the differential considerations to make an early diagnosis and to start an adequate therapy early. PMID- 9849053 TI - [Recurrent tetany as the first symptom of late manifesting celiac disease]. AB - BACKGROUND: The diagnostic classification of a malabsorption is often made more difficult, particularly in oligosymptomatic forms. CASE REPORT: A female patient is presented, where the diagnosis of malabsorption syndrome was concluded because of recurring tetanias. This could be traced back to an oligosymptomatic celiac disease. Diagnostic course of action, differential diagnosis as well as further observation during therapy are discussed. CONCLUSION: Oligosymptomatic developments of celiac disease are common amongst adults and present a diagnostic challenge. The existence of a malabsorption should be considered, even if the momentary individual symptoms are unclarified. PMID- 9849054 TI - [Multi-organ involvement in recurrent Moschcowitz disease. Splenectomy as ultima ratio in a therapeutic dilemma?]. AB - BACKGROUND: The symptoms primarily described in Morbus Moschcowitz included thrombocytopenic purpura and hemolytic anemia. In addition it presents with a variety of clinical manifestations depending on the organs involved (i.e. neurological, renal, gastrointestinal, cardiac involvement). It is a rare disease and the pathogenesis still remains unclear. The efficacy of derived therapeutical concepts can hardly be assessed in controlled trials. CONCLUSION: Currently the main option seems to be plasma therapy. In non-responders surgical procedures (splenectomy) may be of benefit to the patient. PMID- 9849055 TI - [Primary biliary cirrhosis: diagnosis and therapy]. PMID- 9849056 TI - [Hematemesis and cholestasis]. PMID- 9849057 TI - [Janus head manifestation of medical-technical progress]. PMID- 9849058 TI - Smoking during pregnancy, 1990-96. AB - OBJECTIVE: This report presents trend data for smoking during pregnancy. Data are presented for various characteristics including age of mother, race and ethnic origin, place of birth of mother, and State of residence. METHODS: Descriptive tabulations were calculated using data reported on birth certificates between 1990 and 1996. RESULTS: The rate of smoking during pregnancy has declined each year between 1990 and 1996. In 1996 over 400,000 women reported smoking during their pregnancies (13.6 percent of all births). The rates of smoking during pregnancy for women 15-19 years of age declined between 1990 and 1994 but increased in the last two years, and now they have the highest rates of all age groups. Among race and ethnic groups, American Indian, non-Hispanic white, and Hawaiian women had the highest rates of smoking during pregnancy in 1996, while Chinese women had the lowest rates. PMID- 9849059 TI - [42nd Annual meeting of the Japanese Society for Medical Mycology. Tokyo, October 25-26, 1998. Abstracts]. PMID- 9849060 TI - [The role of vaccination in the prevention of communicable diseases]. AB - The increasing role of vaccination in prevention of infectious diseases, the newly introduced vaccines and immunization systems are dealt with. Some recent and future alterations in the field of the basic vaccination system are discussed. Special emphasis is laid on the new vaccines and immunization methods which can be used with good results in preventing some important infectious diseases at home, as well as staying in developing countries, too. PMID- 9849061 TI - [Endoscopic therapy of pancreas divisum]. AB - Pancreas divisum is the most frequent congenital ductal anomaly of the pancreas: it occurs in 5-10% of the population. In the majority of patients, this congenital anomaly is of no clinical importance. In a certain subset of patients, however, pancreas divisum is clinically important as a cause of abdominal pain, acute recurrent pancreatitis or chronic obstructive pancreatitis. The authors, experience on endoscopic drainage of the minor papilla is reported. In the history of patient 1., three episodes of recurrent pancreatitis and permanent upper abdominal pain were explored. ERP revealed a pancreas divisum and a mild irregularity and dilation of the dorsal pancreatic duct. A 7 F stent (length: 6 cm) was implanted in the dorsal pancreatic duct following a papillotomy on the stenotic minor papilla. A repeated Lundh test revealed a 58% improvement in the exocrine pancreatic function. No recurrence of pancreatitis has been observed in spite of the moderate continuous abdominal pain. In patient 2., ERP demonstrated a pancreas divisum and a severely dilated dorsal pancreatic duct as causes of the previous permanent abdominal pain. An 8 F stent (length: 5 cm) was inserted through the minor papilla without endoscopic sphincterotomy. A significant improvement in exocrine pancreatic function (70%) ensued. No abdominal pain has since been observed. In conclusion, dorsal pancreatic duct stenting (mainly in cases involving a dilated pancreatic duct) seems to have a beneficial effect in patients with both recurrent acute pancreatitis or chronic obstructive pancreatitis evoked by pancreas divisum. PMID- 9849062 TI - [Chlamydia trachomatis infections]. AB - Chlamydia trachomatis is the most common bacterial cause of sexually transmitted disease in the developed countries. The most important implications of genital chlamydia infections involve the reproductive health sequalae of upper genital tract infections in women: pelvic inflammatory disease, ectopic pregnancy, and infertility, infection by this organism is insidious, symptoms are absent or minor among most infected women and many man. This large group of asymptomatic and infectious persons sustains transmission within a community. This review discusses the epidemiology, of sexually transmitted chlamydial infections, the spectrum of clinical manifestations and their sequelae, the laboratory diagnosis of genital chlamydiasis, antibiotic treatment and prevention. PMID- 9849063 TI - [Familial heterotaxy syndrome. Case report and review of the international literature]. AB - The authors report two families with two affected siblings of heterotaxy syndrome. Ivemark syndrome with asplenia and complex cardiovascular malformation occurred in two siblings of the first family. The first affected sibling in the second family had situs inversus, transposition of the great arteries with spleen on the right side of the abdomen. Ivemark syndrome with polysplenia and cardiovascular malformation were present in the second affected sibling of the second family. Autosomal recessive inheritance of Ivemark syndrome was reported in the most of the cases, but there are several cases of autosomal dominant inherited Ivemark syndrome. X-linked inheritance of heterotaxy syndrome is also known. Heterotaxy syndromes could also occurred in chromosomal translocation or deletion in sporadic cases. The molecular genetic studies were not able to find the mutation responsible for heterotaxy syndrome. The diagnosis of heterotaxy syndrome could be made by foetal echocardiography until molecular genetic methods are available. Therefore, in the case of positive anamnesis, foetal echocardiography on the 18-20 weeks of gestation is essential diagnostic method. PMID- 9849064 TI - [Ventricular tachycardia masquerading as supraventricular tachycardia]. AB - To distinguish supraventricular tachycardia with aberrancy from ventricular tachycardia is sometimes difficult. It seems to be easy to distinguish the two forms in patients with preexisting bundle branch block: if the QRS morphology during tachycardia is identical to those during at rest the tachycardia is supraventricular, if different, ventricular. We present two cases with preexisting bundle branch block and wide complex tachycardia whose QRS morphologies were almost same to those during normal rest rhythm. The atrioventricular dissociation and the response to adenosine and lidocaine strongly suggests ventricular tachycardia. In these cases ventricular tachycardia masqueraded as supraventricular tachycardia and the identical QRS morphology with the preexisting bundle branch block may suggest a misdiagnosis of supraventricular tachycardia. PMID- 9849065 TI - [Decline ot the old municipal hospital in Pest and the preliminaries for the establishment of the Rokus Hospital]. PMID- 9849066 TI - [Hungarian Medical Journal 100 years ago: Dentistry at the time of Hippocrates]. PMID- 9849067 TI - [Separate hospital department for patients with gastrointestinal hemorrhage]. PMID- 9849068 TI - Foscarnet. PMID- 9849069 TI - Fever. AB - Fever is caused by a resetting of the thermoregulatory center in the hypothalamus, most often by viral or bacterial infections. The specific etiologic diagnoses vary, depending on the child's age and height and duration of fever. Fever lasting 7 days or more is termed an FUO. When evaluating children who have fever, the most critical diagnostic maneuver is the carefully performed clinical evaluation, observation, history, and physical examination. Laboratory studies may be ordered based on the results of the clinical evaluation as well as the child's age and height and duration of the fever. Recent studies have outlined the manner in which data from the clinical evaluation and laboratory test results may be integrated to formulate an effective plan of management and follow-up. PMID- 9849070 TI - Providing effective reproductive health care and prescribing contraceptives for adolescents. PMID- 9849071 TI - Chronic diarrhea. AB - The differential diagnosis of chronic diarrhea varies markedly with age. In infants, it is usually a problem with formula intolerance. Because there is up to a 50% crossover intolerance between milk and soy, the infant should be given an extensively hydrolyzed formula. If such intervention is delayed, he or she may develop intractable diarrhea of infancy. Most affected toddlers have either irritable colon of infancy or protracted viral enteritis with low-grade mucosal injury and are consuming hypertonic feedings. In either case, institution of a high-fat, low-carbohydrate diet that includes whole milk often results in significant improvement. Dietary lactose rarely is a problem. A likely cause of diarrhea among children and adolescents is inflammatory bowel disease. With the exception of toddlers, chronic diarrhea suggests the presence of significant organic disease. PMID- 9849072 TI - Clinical pharmacology and therapeutic drug monitoring in neonates and children. AB - From a pharmacotherapy perspective, the process of development and growth represents an unstable and dynamic condition. Age-related changes in drug absorption, distribution, and metabolism among neonates, infants, and prepubescent children create a unique situation that may increase drug toxicity of some agents and protect from toxicity of other agents. Understanding the age related changes in drug disposition that are relevant for therapeutic response and toxicity is essential for optimizing pharmacotherapy at different stages of childhood. PMID- 9849073 TI - Index of suspicion. Case 1. Diagnosis: chronic acalculous cholecystitis. PMID- 9849074 TI - Index of suspicion. Case 2. Diagnosis: bacillary dysentery and campylobacter infections. PMID- 9849075 TI - Index of suspicion. Case 3. Diagnosis: hematemesis resulting from gastritis, duodenal ulcer and hemophilia A. PMID- 9849076 TI - Bonding by Kennell & Klaus. PMID- 9849077 TI - Type 2 diabetes in adolescents, no longer rare. PMID- 9849078 TI - [A complex approach to protecting the health of military missile personnel (an interview of Colonel General V. N. Iakovlev, Commander-in-Chief of the Strategic Missile Forces). Interview by N. F. Shalaev]. PMID- 9849079 TI - [The system of psychoprophylactic measures and psychiatric care in the Strategic Missile Forces]. AB - Since 1992 psychiatric morbidity, hospitalisation and discharge rates among the SRF personnel show a tendency to increase, reaching accordingly 7.7/1000, 7.3/1000 and 2.7/1000. The existing psychoprophylaxis programme includes primary medical and psychoexamination of young recruits, medical check-ups for servicemen going on duty and on guard, sociometric and psychological studies of the servicemen interpersonal relations with the aim of correction thereof and medical control of those who return to the troops after leaves or temporary assignments. The SRF's medical service closely coordinates its actions with the Ministry of Health's appropriate setups, commanding officers and personnel instructors. PMID- 9849080 TI - [The organizational characteristics of surgical care in the Strategic Missile Forces]. PMID- 9849081 TI - [The organization of the management system and the means for improving therapeutic care in the Strategic Missile Forces]. PMID- 9849082 TI - [Medical support for the cleanup of the aftermath of accidents and catastrophes in the Missile Forces]. PMID- 9849083 TI - [The use of transosseous osteosynthesis in treating fractures of the tubular bones]. PMID- 9849084 TI - [The treatment of paranasal sinusitis]. PMID- 9849085 TI - [Improvement in cardiological care in the Strategic Missile Forces]. PMID- 9849086 TI - [The effect of the laser irradiation of the blood on its lipid and phospholipid components in diabetes mellitus]. AB - Study results of the laser irradiation impact on the blood lipid and phospholipid components and membrane erythrocyte in patients with diabetes mellitus and pyo necrotic injuries of lower extremities when treated by laser intravenously or epicutaneously. The results prove, that the laser irradiation of low intensity favourably affects the blood components in a complex treatment of diabetes mellitus. Intravenous irradiation positive dynamics are more easily observed. PMID- 9849087 TI - [Experience in developing an automated work place for the endoscopist]. PMID- 9849088 TI - [Modern practical syphilology]. PMID- 9849089 TI - [Measures to prevent endometrial cancer in hyperplastic processes]. PMID- 9849090 TI - [The treatment of middle-aged and elderly patients suffering from trochanteric fractures of the femur]. AB - Treatment of 198 elderly patients with trochanteric fractures included different conservative and operative therapy methods. For the first time in Russia a new approach of extrafocus osteosynthesis was recommended for "decompensated" patients. It is proved, that for this category of the injured, active surgery tactics results in a considerable improvement of treatment outcome. PMID- 9849091 TI - [Treatment experience with closed injuries to the bursal-ligamental apparatus of the knee joint]. PMID- 9849092 TI - [The prospects for the use of millimeter-range electromagnetic radiation in duodenal peptic ulcer]. PMID- 9849093 TI - [The organizational characteristics of health and hygiene and epidemic-control measures in the Strategic Missile Forces]. PMID- 9849094 TI - [The hygienic characteristics of the medical technology accompaniment to the development, creation and operation of installations equipped with video display terminals]. AB - Radiation measuring on the work places of operators in command and control installations proved that environment parameters depending on electronic display functioning are in line with the regulations' requirements. Nevertheless the operator health estimates show that the problem of personnel security still exists. The authors recommend some measures to improve the situation. PMID- 9849095 TI - [The 80th anniversary of the Epidemiological Health Unit of the Leningrad Military District]. PMID- 9849096 TI - [The 60th anniversary of the Epidemiological Health Unit of the Northern Fleet]. PMID- 9849097 TI - Infants' expectations about object label reference. AB - The aim of this research was to examine whether infants at the early stages of lexical development were sensitive to the word-category linkage. In Experiment 1, 16-to 19-month-old infants were requested to match a target with either a basic level or a thematic match, with or without a novel label. Stimuli were presented using the preferential looking paradigm. Infants in the Novel Label condition looked significantly longer at the basic-level match than infants in the No Label condition. In Experiment 2, infants were presented with a target, followed by a basic-level match and a superordinate-level match with or without a novel label. Again, infants in the Novel Label condition looked significantly longer at the basic-level match than infants in the No Label condition. Taken together, these findings indicate that infants initially assume that novel words label basic level categories and thereby do honour the word-category linkage. PMID- 9849098 TI - Adult age differences in on-line contingency judgements. AB - This research was designed to further our understanding of age differences in contingency judgements and to consider the role of working memory in such judgements. Experiment 1 examined age differences in contingency judgement accuracy when information processing requirements were manipulated. Young and older adults estimated the degree of contingency between two categorical variables. Contingencies, amount of information, and speed of execution were varied. Participants discriminated the contingencies well. Judgement accuracy declined with smaller numbers of trials and shorter intertrial intervals. The judgement deterioration was more severe for older adults. In Experiment 2, speed interfered again more with the judgements made by older adults. Modulations of this effect occurred as a function of stimulus duration and distribution of event frequencies. Overall, the judgement deterioration is consistent with Wagner's (1981) model of automatic memory processing. PMID- 9849099 TI - Telepresence. AB - Telepresence, the perception of presence within a physically remote or simulated site, has been identified as a design ideal for synthetic environments. However, confusion exists within the literature about the precise definition of telepresence. Furthermore, there is a need for a plausible and parsimonious model of telepresence. This paper identifies three types of telepresence extant in the literature: simple telepresence, cybernetic telepresence, and experiential telepresence. The third definition is the most interesting. This paper reviews the origins of experiential telepresence and the theoretical approaches commonly used to explain it. One can term these technological approaches, which emphasize the role of control/display technology, and psychological approaches, which identify experiential telepresence with known psychological phenomena. Finally, the paper presents and discusses an integrative approach to telepresence featuring a structured attentional resource model. Actual or potential applications of this research include the design of future human-machine interfaces for teleoperated robots and virtual reality systems. PMID- 9849100 TI - Ocular vergence measurement in projected and collimated simulator displays. AB - The purpose of this study was to investigate electrooculography (EOG) as a measurement of ocular vergence in both collimated and projected simulator environments. The task required participants to shift their gaze between a central fixation point and a target appearing at one of three eccentricities. EOG was effective in recording ocular vergence. The EOG results were similar between collimated and projected displays, except for differences in vergence changes during lateral movement of the eyes, and ocular excursions downward elicited a greater EOG response than the reverse upward movement. The computer-based technique of recording vergence was found to produce measurable traces from a majority of participants. The technique has potential for further development as a tool for measuring ocular vergence in virtual environments where methods that require the wearing of head-mounted apparatus to track ocular structures (e.g., the pupil), which cannot be worn at the same time as a flight or flight-simulator helmet, are unsuitable. PMID- 9849101 TI - Empirical models based on free-modulus magnitude estimation of perceived presence in virtual environments. AB - A series of 3 studies was conducted to test free-modulus magnitude estimation as a measure of perceived presence in virtual environments (VEs) and to model the first- and second-order effects of 11 VE system parameters on perceived presence across 5 subtasks. Sequential experimentation techniques were used to build 4 empirical models using polynomial regression. An integrated empirical model of data combined across 2 experiments demonstrated that all significant factors had a positive effect on perceived presence. Three of these parameters--field of view, sound, and head tracking--had almost 3 times as much influence on presence than the other 4 significant parameters, which were visual display resolution, texture mapping, stereopsis, and scene update rate. Sequential experimentation was an efficient tool for building empirical models of perceived presence, but the subjective nature of this phenomenon and individual differences made data bridging across sequential studies problematic. It was concluded that magnitude estimation is a useful measure of perceived presence, and the resulting polynomial regression models can be used to facilitate VE system design decisions. This research has broad application in the selection and design of VE system components and overall design of VE systems. PMID- 9849102 TI - Effects of variation in system responsiveness on user performance in virtual environments. AB - System responsiveness (SR) is defined as the elapsed time until a system responds to user control. SR fluctuates over time, so it must be described statistically with mean (MSR) and standard deviation (SDSR). In this paper, we examine SR in virtual environments (VEs), outlining its components and methods of experimental measurement and manipulation. Three studies of MSR and SDSR effects on performance of grasp and placement tasks are then presented. The studies used within-subjects designs with 11, 12, and 10 participants, respectively. Results showed that SDSR affected performance only if it was above 82 ms. Placement required more frequent visual feedback and was more sensitive to SR. We infer that VE designers need not tightly control SDSR and may wish to vary SR control based on required visual feedback frequency. These results may be used to improve the human-computer interface in a wide range of interactive graphical applications, including scientific visualization, training, mental health, and entertainment. PMID- 9849104 TI - Aurally and visually guided visual search in a virtual environment. AB - We investigated the time participants took to perform a visual search task for targets outside the visual field of view using a helmet-mounted display. We also measured the effectiveness of visual and auditory cues to target location. The auditory stimuli used to cue location were noise bursts previously recorded from the ear canals of the participants and were either presented briefly at the beginning of a trial or continually updated to compensate for head movements. The visual cue was a dynamic arrow that indicated the direction and angular distance from the instantaneous head position to the target. Both visual and auditory spatial cues reduced search time dramatically, compared with unaided search. The updating audio cue was more effective than the transient audio cue and was as effective as the visual cue in reducing search time. These data show that both spatial auditory and visual cues can markedly improve visual search performance. Potential applications for this research include highly visual environments, such as aviation, where there is risk of overloading the visual modality with information. PMID- 9849103 TI - Effects of localized auditory information on visual target detection performance using a helmet-mounted display. AB - An experiment was conducted to evaluate the effects of localized auditory information on visual target detection performance. Visual targets were presented on either a wide field-of-view dome display or a helmet-mounted display and were accompanied by either localized, nonlocalized, or no auditory information. The addition of localized auditory information resulted in significant increases in target detection performance and significant reductions in workload ratings as compared with conditions in which auditory information was either nonlocalized or absent. Qualitative and quantitative analyses of participants' head motions revealed that the addition of localized auditory information resulted in extremely efficient and consistent search strategies. Implications for the development and design of multisensory virtual environments are discussed. Actual or potential applications of this research include the use of spatial auditory displays to augment visual information presented in helmet-mounted displays, thereby leading to increases in performance efficiency, reductions in physical and mental workload, and enhanced spatial awareness of objects in the environment. PMID- 9849105 TI - The influence of body movement on subjective presence in virtual environments. AB - We describe an experiment to assess the influence of body movements on presence in a virtual environment. In the experiment 20 participants were to walk through a virtual field of trees and count the trees with diseased leaves. A 2 x 2 between subjects design was used to assess the influence of two factors on presence: tree height variation and task complexity. The field with greater variation in tree height required participants to bend down and look up more than in the lower variation tree height field. In the higher complexity task participants were told to remember the distribution of diseased trees in the field as well as to count them. The results showed a significant positive association between reported presence and the amount of body movement in particular, head yaw--and the extent to which participants bent down and stood up. There was also a strong interaction effect between task complexity and gender: Women in the more-complex task reported a much lower sense of presence than in the simpler task. For applications in which presence is an important requirement, the research in this paper suggests that presence will be increased when interaction techniques are employed that permit the user to engage in whole body movement. PMID- 9849106 TI - Contributions of proprioception to navigation in virtual environments. AB - Users immersed in virtual environments (VEs) are prone to disorientation and have difficulty transferring spatial knowledge to the real world. A single experiment investigated the contribution of inadequate proprioception to this problem by providing participants with interfaces to a virtual environment that either did (a walking interface) or did not (a joystick) afford proprioceptive feedback similar to that obtained during real walking. The 2 groups explored a large, complex building using a low-resolution head-mounted display. Later, their navigational abilities within the actual building were compared with those of control groups who either studied a map of the building, walked through the real building, or received no prior training. The walking interface conveyed no benefit on an orientation task performed during training in the VE, but it did benefit participants when they tried to find objects in the real world. Actual or potential applications include simulations of environments that are normally explored on foot but cannot be readily visited, such as infantry battlefields and facilities contaminated with chemical, biological, or radiological materials. PMID- 9849107 TI - Evidence from a partial report task for forgetting in dynamic spatial memory. AB - G. Sperling (1960) and others have investigated memory for briefly presented stimuli by using a partial versus whole report technique in which participants sometimes reported part of a stimulus array and sometimes reported all of it. For simple, static stimulus displays, the partial report technique showed that participants could recall most of the information in the stimulus array but that this information faded quickly when participants engaged in whole report recall. An experiment was conducted that applied the partial report method to a task involving complex displays of moving objects. In the experiment, 26 participants viewed cars in a low-fidelity driving simulator and then reported the locations of some or all of the cars in each scene. A statistically significant advantage was found for the partial report trials. This finding suggests that detailed spatial location information was forgotten from dynamic spatial memory over the 14 s that it took participants to recall whole report trials. The experiment results suggest better ways of measuring situation awareness. Partial report recall techniques may give a more accurate measure of people's momentary situation awareness than whole report techniques. Potential applications of this research include simulator-based measures of situation awareness ability that can be part of inexpensive test batteries to select people for real-time tasks (e.g., in a driver licensing battery) and to identify people who need additional training. PMID- 9849108 TI - The relationship between low back discomfort ratings and the NIOSH lifting index. AB - An epidemiological study was performed to evaluate the relation between low-back discomfort ratings and use of the revised NIOSH lifting guide to assess the risk of manual material handling (MMH) tasks. We surveyed 97 MMH workers on site in 15 factories and designed a questionnaire to systematically collect job-related information. Approximately 90% of the workers had suffered various degrees of lower back discomfort, and 80% had sought medical treatment. The survey showed that 42 of the 97 jobs analyzed had a recommended weight limit of 0, which was attributed to either a horizontal distance or a lifting frequency that exceeded the bounds of the NIOSH lifting index. Apparently, the limits for horizontal distance and maximum allowable frequency are too stringent to accommodate many existing MMH jobs. For the remaining 55 jobs, the significant positive correlation obtained between the lifting index and the severity of low-back discomfort suggests that the lifting index is reliable in assessing the potential risk of low-back injury in MMH. These findings provide useful information on the application of the NIOSH lifting guide to the assessment of low-back pain. PMID- 9849109 TI - Measuring skill acquisition and retention with an ATM simulator: the need for age specific training. AB - The present study focused on the type of information presented during training and its effects on initial and retention performance of older and younger adults interacting with computerized, new technology. The effects of emphasizing concepts versus actions during training on performance immediately after training and after a 1-month retention interval were examined. Younger and older adults completed either action or concept training for operating a virtual automatic teller machine (ATM). Overall, action training was associated with faster and more accurate performance immediately after training and better retention performance for older adults. For older adults, value of type of training interacted with type of task component. These findings are applicable to the development of age-specific training materials for computerized tasks. PMID- 9849110 TI - Meta-analyses comparing theory of mind abilities of individuals with autism, individuals with mental retardation, and normally developing individuals. AB - A deficit in theory of mind (ToM) abilities has been described as the core deficit in autism. The authors performed 3 meta-analyses, comparing ToM abilities of individuals with autism, individuals with mental retardation (MR), and normally developing individuals. Results indicated that individuals with autism and MR have impaired ToM abilities. The etiology associated with MR (i.e., Down syndrome, undifferentiated etiology) was found to be an important moderator variable. Chronological age (CA) and verbal mental age (VMA) of the normally developing children and CA, VMA, and performance mental age of individuals with MR, and type of matching between the groups were also found to be moderator variables. Discussion focuses on the implication of the findings and emphasizes the need to consider the specific etiology of comparison groups when studying abilities and impairments of individuals with autism and MR. PMID- 9849111 TI - How shall we speak of children's personalities in middle childhood? A preliminary taxonomy. AB - Developmental researchers have neglected the study of personality traits in middle childhood, thus leaving unanswered many questions about childhood personality structure. This article presents a developmental framework for understanding personality in middle childhood and critically reviews 5 models of temperament and personality structure in this age range: the models of A. Thomas and S. Chess, A. H. Buss and R. Plomin, M. K. Rothbart, J. Block and J. H. Block, and the Big Five. A number of robust personality dimensions common to these models and the broader developmental and adult personality literatures are then discussed: sociability, social inhibition, dominance, negative emotionality, aggressiveness, prosocial disposition, persistence/attention, mastery motivation, inhibitory control, and activity level. These dimensions represent a preliminary taxonomy of personality traits for exploring questions of individual development in childhood. PMID- 9849112 TI - Eye movements in reading and information processing: 20 years of research. AB - Recent studies of eye movements in reading and other information processing tasks, such as music reading, typing, visual search, and scene perception, are reviewed. The major emphasis of the review is on reading as a specific example of cognitive processing. Basic topics discussed with respect to reading are (a) the characteristics of eye movements, (b) the perceptual span, (c) integration of information across saccades, (d) eye movement control, and (e) individual differences (including dyslexia). Similar topics are discussed with respect to the other tasks examined. The basic theme of the review is that eye movement data reflect moment-to-moment cognitive processes in the various tasks examined. Theoretical and practical considerations concerning the use of eye movement data are also discussed. PMID- 9849113 TI - The role of temporal landmarks in autobiographical memory processes. AB - The prevalence of temporal landmarks has been revealed in a wide range of tasks associated with autobiographical memory, such as recall and dating of personal events. The author examined 3 types of events that have usually been considered landmarks: flashbulb memories, 1st experiences, and reference points in personal histories. The differences between landmark events and nonlandmark events are considered, and evidence is presented showing the effect of these landmarks on memory tasks. It is suggested that the effect of temporal landmarks on these tasks is twofold: (a) performance on the tasks is intricately linked with how our autobiographical memory system is structured, and temporal landmarks may play a critical role in this organization; and (b) landmarks may be actively and spontaneously used by individuals as cues when performing recall and dating tasks (the cuing hypothesis). Both of these factors serve to systematically bias performance, leading to robust phenomena such as calendar effects and reminiscence peaks in recall and improvements in accuracy in dating. PMID- 9849114 TI - The economics of breastfeeding in Singapore. AB - A study of 340 mothers was conducted in Kandang Kerbau Hospital on September 1992 to determine if it were more economical for households to breastfeed or bottle feed an infant for the first three months. Two economic models, a low cost model and a high cost model, were adopted incorporating a mathematical expression from Almroth's work in 1979. The savings in a mother's gross income for the period ranged from 3% to 9% for the low cost model and from 8% to 21% for the high cost model. From the household perspective, two components contributed to the economic savings of breastfeeding over artificial feeding: the cost of goods consumed and the time taken to feed the baby. It was noted that the time taken to artificially feed is longer than the time taken to breastfeed an infant. The results of this study provided more concrete basis for policy makers and advocates of breastfeeding to promote breastfeeding in Singapore. The amount of savings from breastfeeding could be considered for the health care system from the public perspective. PMID- 9849115 TI - Developing effective interactions to improve breastfeeding outcomes. AB - The objective of this replicated research was to assess the effectiveness of antenatal Breastfeeding Workshops (Jamieson 1990) in improving breastfeeding outcomes. It was a longitudinal study using a pre-test/post-test design to evaluate any increase in the confidence, skill and knowledge of women and midwives. The research was carried out in the maternity section of a large Tasmanian teaching hospital. Fifty-six midwives who attended the workshops became the research group with 49 non-attendees as the control. Of the 159 women who indicated an interest in attending, 107 who attended formed the research group, and the remaining 52 were used as the control. The paper is in two parts. The first describes responses from midwives. The results show those midwives who attended a workshop perceived an increase in their confidence level in supporting mothers to breastfeed, altered their perception of those factors most important in successful breastfeeding, and acknowledged that updating of breastfeeding knowledge is necessary on an annual basis. The second part analyses breastfeeding outcomes in respondent women. The results show that women who attended the workshops had a significant increase in confidence levels associated with breastfeeding, and were more likely (p < 0.01) to breastfeed for an extended time even though they experienced difficulties. PMID- 9849116 TI - Breastfeeding a baby with gastric reflux. PMID- 9849117 TI - Breastfeeding and the use of recreational drugs--alcohol, caffeine, nicotine and marijuana. AB - This paper both reviews the current literature and explores anecdotal information as reported by Nursing Mothers' Breastfeeding Counsellors relating to breastfeeding and the use of alcohol, caffeine, nicotine and marijuana. All of these drugs do enter breastmilk to some extent and can have a detrimental effect on the production, volume, composition and ejection of breastmilk, as well as a direct adverse effect on the infant. Breastfeeding mothers should be encouraged to restrict their intake of these so-called recreational drugs. It is acknowledged that this is a particularly stressful period in a mother's life and that she may need additional support and practical suggestions to limit the exposure of these drugs to the infant. PMID- 9849118 TI - The engorgement enigma. AB - A search of the literature reveals only a few studies that deal directly with engorgement. When the relevant research is analysed, a picture emerges of the causes of engorgement, how it can be prevented and what is the best management when it occurs. Equipped with this information, people providing support to breastfeeding mothers can encourage preventative behaviours, and assess and educate mothers at risk, thus allowing the majority of mothers and babies to breastfeed naturally without intervention. PMID- 9849119 TI - Breaking the silence. PMID- 9849120 TI - Medicare brings new challenges in home health care. PMID- 9849121 TI - Taking a risk on a dream job. PMID- 9849122 TI - Urinary Incontinence in Women. AB - Despite the prevalence of urinary incontinence, most affected women don't seek help, primarily because of embarrassment or because they are not aware that effective treatment is available. Failure to store urine may be caused by an abnormality in any component of the lower urinary tract. Common abnormalities are poor bladder compliance and bladder outlet failure. Patients who experience failure to empty can present with recurrent urinary tract infections, retention or incontinence. Using a symptom-based classification of incontinence, this would be referred to as overflow incontinence. Other possible categories of urinary incontinence are failure to store and empty and functional incontinence. A combination of a failure to store and empty is difficult to diagnose and treat clinically. Treatments are directed at the particular cause of incontinence and can include medical or surgical therapies. PMID- 9849123 TI - Acute diarrhea. AB - Diarrhea can result from damage to the intestinal lining caused by viruses or bacteria, malabsorption, inflammatory processes, bile salt and pancreatic enzyme deficiency, abnormal motility, or the presence of osmotically active solutes in the gut. While it is important to elicit information to determine the possible cause of diarrhea, be sure to check circulatory status first. Some patients may need rehydration therapy more urgently than they need a diagnosis. The main goals of treatment are to prevent dehydration and correct electrolyte imbalance, to provide supportive and symptomatic therapy, and to treat underlying disease. In most cases, a specific diagnosis is not necessary to guide initial treatment. PMID- 9849124 TI - An introduction to colposcopy. Concepts, controversies and guidelines for practice. AB - Colposcopy is a clinical procedure used to investigate the health of the cervix and lower genital tract. This article is limited to an overview of the colposcopic examination. Colposcopy consists of four sequential steps visualization, assessment, sampling, and documentation and correlation. Since most women are dependent on their primary care providers for information and guidance, a post-colposcopic appointment will aid in the discussion of treatment options and help alleviate any fears or unanswered questions. Regardless of the treatment, stress the need for frequent follow-up exams for Pap smears or repeat colposcopy. PMID- 9849125 TI - The changing face of NSAID therapy. PMID- 9849126 TI - Pressure ulcer management. PMID- 9849127 TI - The number game. Primary care of adolescents with hypertension. PMID- 9849128 TI - At last, the true diagnosis. A case report. PMID- 9849129 TI - It can happen to you. Why you should carry your own malpractice insurance. PMID- 9849130 TI - Interrupting the maternal instinct. Mastitis in breastfeeding women. PMID- 9849131 TI - Advances in cancer research. PMID- 9849132 TI - Profession's leadership needs to address NP glut. PMID- 9849133 TI - Urinary tract infections may trigger relapse in multiple sclerosis. AB - Multiple Sclerosis (MS), a demyelinating disease of the central nervous system, is the most common neurological disease affecting young adults in North America and, in the majority of cases, is associated with accumulating disability. Urinary tract dysfunction affects up to 90% of the MS population, and urinary tract infections are encountered in up to 74% of the tested population. Viral infections have previously been shown to trigger acute exacerbation and it is our experience that urinary tract infection also commonly precedes relapse, and, when recurrent, is associated with neurologic progression. We present three case studies from our MS Clinic where recurrent UTI was associated with acute exacerbation and neurologic progression refractory to intravenous steroid treatment. Interferons, protein signaling molecules, have recently been found to play a role in acute exacerbation and disease progression in individuals with MS. Viral infections induce interferon release which may activate T cells to produce gamma-interferon. Interferon-gamma precipitates relapse and stimulates production of tumour necrosis factor-alpha, a cytokine directly toxic to oligodendrocytes. Bacterial infections similarly induce interferon release and may activate immune pathways that result in MS exacerbation and neurologic progression. PMID- 9849134 TI - On matters not measured. A condensed version of the Mary Glover Lecture. PMID- 9849135 TI - Vagus nerve stimulation therapy: nurses role in a collaborative approach to a program. AB - Approximately 300,000 Canadians have epilepsy. Of those 30 percent fail to achieve satisfactory seizure control with current antiepileptic drug therapy (Vagus Nerve Stimulator Study Group, 1995). The development and availability of new therapeutic options cannot be overlooked for medically intractable patients. Chronic Vagus Nerve Stimulation (VNS) has demonstrated a 50 percent reduction in seizure frequency in 1/3 of patients with refractory partial onset seizures (Uthman, et al, 1993). Individuals undergoing this procedure require the attention of health care professionals from both the neurological and neurosurgical programs. This unique intervention demands that the patient's device be tested intra-operative, and programming begin during the immediate post operative phase. Assessment of tolerance and side effects to vagus nerve stimulation therapy, as well as continued evaluation of the patients seizure control are necessary to direct staged programming of the device. This poster will demonstrate how the nurses from the neurology and neurosurgery clinics have been able to collaborate to ensure patients needs are met. Patient education is crucial to assisting the patient through this procedure, and key points will be identified. The implementation of coordinating the approach for programming the patient's device will be depicted. Future recommendations for long-term outcome measurement will be addressed. PMID- 9849136 TI - Nursing research in stroke. A review. AB - Nurses have demonstrated an interest in research in stroke. Studies encompass a broad variety of subjects; some clearly within the traditional realm of nursing and others that are common to multiple disciplines. Studies are limited in number and most lack the necessary numbers or controls that make data meaningful. Important beginnings are evident. Research about nurses who care for stroke survivors attempts to find correlations between attitudes and knowledge or outcome measures. Care delivery systems and models of nursing practice may impact on a variety of outcome measures, both those which are patient centred (functional status, quality of life) and those which are system driven (length of stay, recidivism). Information regarding the experience of stroke is rich in detail but limited in quantity. Much more information must be gained from a broader segment of the population to provide a baseline of understanding. Similarly, caregiver research begins to touch on the experience but considerable variation must exist between spouse caregivers and adult children or others, and between cultures. Finally, our interventions and their effect on outcomes are only just beginning to be studied. We remain a great distance from our ideal "evidence based practice". On a positive note, the research that nurses have conducted within the field of stroke is indicative of the broad interests that exist. To provide the care that is needed for our aging population while resources shift and shrink, it is essential that we not only test our interventions and their impact on outcomes but that we also are prepared to blur the traditional professional boundaries. Nurses do not have a monopoly on family centred care and care for the caregivers, nor do we act in isolation when we implement strategies to normalize bowel function post stroke. Collaborative research between disciplines and across cultural and political barriers is essential; resources for funding are available but have not been successfully accessed. The time is now for research by nurses and colleagues to begin to bridge the many knowledge gaps that persist. PMID- 9849137 TI - Does nursing care make a difference? PMID- 9849138 TI - The continuum of care: the process and development of a nursing model for stroke education. AB - A recent focus in health care is to develop a seamless transition for patients between the variety of health care agencies with whom they interact. The continuum of care for patients with stroke is complex and many levels of care are required, from acute intervention through to rehabilitation and reintegration into the community. A group of nurses from a variety of agencies in Southwestern Ontario received funding from the Ontario province Wide Nursing Project to effect a desirable change in the way nurses care for the stroke patient. This article describes the Ontario Province Wide Nursing project and the process of and challenges in developing a nursing model for stroke. The development of a nursing model for stroke patients and their families involved conducting nursing, and patient and family focus groups to identify key issues. Thirty-eight nurses participated in the nursing focus groups, and 8 patients and 18 family members participated in the patient and family focus groups. The common themes were the need for improved communication and the need to better prepare patients and families for the many transitions they experience during their recovery from stroke. A Stroke Education Record, which identifies a comprehensive list of potential learning needs of stroke patients and their families, and a Stroke Education and Resource Guide, which provides a reference for nurses, were developed and will be implemented in September 1998. Implementation of the model included a workshop, staff inservices, a self-learning package, and unit displays. Evaluation of the model will include nursing and patient and family focus groups, a chart review, written nursing evaluations, and patient and family telephone interviews. PMID- 9849139 TI - Stroke education: the development of a documentation system and resource guide. PMID- 9849141 TI - Clinical risk management: is it just a sham? PMID- 9849140 TI - Nurses must start writing so as to be understood. PMID- 9849142 TI - Syphilis: signs, symptoms, treatment and nursing management. AB - The rise in the number of cases of syphilis reported over the last 3 years (Communicable Disease Report, 1997) suggests that some individuals are not practising safer sex and/or that health education programmes are ineffective. The increasing incidence could also be a consequence of more people attending for sexual health checks. Syphilis is primarily a sexually transmitted disease, although a pregnant woman may pass the disease to her fetus via the placenta, and rarely healthcare professionals may acquire the disease through open lesions. If allowed to run its full course the disease can have devastating effects on the individual. Regular screening of at-risk groups, screening at antenatal clinics and genitourinary medicine clinics, promotion of safer sex activities and health education may aid prevention. The nurse is ideally placed to provide patients with information that may prevent infection or help to minimize its severity. PMID- 9849143 TI - Development of a community nurse-led continence service. AB - Extrapolations from prevalence studies suggest that on average 56,000 adults experience urinary incontinence in Glasgow, a third of whom will have been incontinent during the last week. A review by a multidisciplinary health gain commissioning team concluded that existing continence services in Glasgow had developed opportunistically and that problems exist, e.g. prescription of products without full assessment of continence problems. In response to this situation, a new community nurse-led continence service was introduced in 1995. This article describes the development and evaluation of this new service. For the past 3 years the service has employed five staff nurses and a physiotherapist. The team is solely employed to promote continence. It carries out assessments both in nursing and residential homes and community clinics. The planned evaluation will assess the effectiveness of this team in promoting continence and the future demand for continence nurse-led services. PMID- 9849144 TI - Role of dopamine in schizophrenia and Parkinson's disease. AB - The neurotransmitter dopamine (DA) and the dopaminergic neurones play an important role in schizophrenia and Parkinson's disease (PD). A decrease in DA in the substantia nigra of the brain has been implicated as the cause of PD. By contrast, it is argued that a functional excess of DA or oversensitivity of certain DA receptors is one of the causal factors in schizophrenia. These factors are reflected in the treatment of both conditions; drugs aimed at increasing DA are prescribed to patients with Parkinson's disease, while most antipsychotic drugs block and reduce the effects of DA. In schizophrenia the antipsychotic effects of traditional 'neuroleptic' drugs such as chlorpromazine are highly correlated with their ability to block DA receptors and reduce the effects of DA. This article examines the role of DA in these two conditions. PMID- 9849145 TI - The implications of head injury for family relationships. AB - Many of those sustaining head injury recover to the point that they no longer require hospital care. However, the family frequently has to cope with an individual with varying degrees of disability involving physical, psychological, cognitive and social dysfunctioning. This review of the literature briefly considers the possible effects that head injury may have for the injured person before going on to discuss the consequences that such an injury may have for the family of that individual. The family's need for information and education is highlighted and it is suggested that the nurse has an important role to play in this context. PMID- 9849146 TI - The link between Down's syndrome and Alzheimer's disease: 2. AB - In this article, the second of two parts, the needs of family and professional carers of people with Down's syndrome and Alzheimer's disease are examined. Substantial numbers of people with Down's syndrome survive to the age of 50 and beyond and so work still needs to be done on finding solutions to the problems faced by this client group and its carers. As well as the difficulties faced by any family carer of a person with dementia, those caring for someone with Down's syndrome and Alzheimer's disease may also have to deal with additional worries and problems. Consideration is given to service provision and the implications for nursing. A case study will illustrate some of the points made. PMID- 9849147 TI - External pneumatic compression therapy for DVT prophylaxis. AB - Thromboembolic disease is a common cause of perioperative death. Emphasis should be placed on prophylactic treatment and therefore this product focus looks at intermittent pneumatic therapy in the form of the Flowtron DVT garment from Huntleigh Healthcare. While anticoagulant therapy is the main prophylactic method for deep vein thrombosis (DVT) the benefits of intermittent pneumatic compression warrant serious consideration. Responsibility for DVT prophylaxis, patient assessment and the monitoring of the effectiveness of the prophylactic therapy lie with the nursing staff. A reliable assessment tool should be used in conjunction with clinical judgment and accurate documentation. PMID- 9849148 TI - Will primary care groups lead to improvements in health care? AB - The author has participated in a series of regional workshops in Wales, Scotland and England where mixed views were expressed by GPs, nurses and primary care professionals regarding the impact of the new White Paper 'The New NHS: Modern, Dependable'. Many welcome the proposals and the opportunity to influence change. Others, however, expressed widespread concern about the proposed changes, particularly the short timescales, lack of skills, resources and information technology with which to achieve the changes, the lack of start-up funds to get the process going and the lack of incentives to sustain it. The input of community nursing to these workshops was disappointing--many were unclear of their role and involvement, and some were waiting for the GPs to involve them; others, however, were starting to set up their own groups to ensure that their views were heard and included. Primary care groups need active involvement of community nurses at management and operational level and nurse representation at governing body level. It is essential for nurses and nursing to get their act together in relation to the decision-making; configuration and determination of the functionality of primary care groups/trusts. PMID- 9849149 TI - The relationship between caring and nursing. PMID- 9849150 TI - Nursing is not a very attractive career option. PMID- 9849151 TI - Negligence is costing the NHS 2.3 billion pounds. PMID- 9849152 TI - Nursing aspects of pressure sore prevention and therapy. AB - Pressure sores remain a significant problem in hospitals and domestic settings, affecting people of all ages, social class and race. Associated complications may be life threatening, e.g. sepsis and osteomyelitis. Other less dangerous, but nevertheless compromising outcomes such as pain, discomfort and low self-esteem and body image can cause personal suffering, and may add extra demand for limited resources. The exact state of pressure sore occurrence remains difficult to determine, particularly in the community. Recent trends in pressure area management present a multidisciplinary approach, eroding traditional perceptions of pressure sores as a solely nursing problem. Written from nursing perspective, this article summarizes principles of good practice relating to pressure sore prevention and therapy, emphasizing the importance of documenting observed events, rather than assumptions or opinions, and the need for healthcare professionals to approach problems and needs from a collaborative stance. Pressure sore risk assessment and classification are discussed, and an overview of nutrition, moving a handling, selecting support surfaces, principles of wound management, and skin care are considered. PMID- 9849153 TI - A policy statement on the prevention of pressure ulcers from the European Pressure Ulcer Advisory Panel. PMID- 9849154 TI - Interventions for confusion and dementia. 1: Quality of life. AB - This series of articles examines some of the more commonly used interventions for confusional states/dementia. This article explores issues around quality of life. Quality of life is a subjective and potentially undefinable concept. Yet in recent years it has been used widely in the nursing literature. However it is defined, it is fundamental to humanity and underpins all the interventions discussed in the later articles. When continuing health care fails to offer hope of a medical cure, the focus necessarily shifts to care. Good care and quality of life are inextricably linked. This article raises awareness of some key issues to guide readers when deciding whether or not to adopt a particular intervention; a prescriptive approach to quality of life is deliberately avoided. Later articles in this series will discuss reality orientation, reminiscence, validation, and multisensory environments. PMID- 9849155 TI - Substance use and mental health problems: meeting the challenge. AB - Dual diagnosis is a term used to describe co-existing substance misuse and mental health problems. Although mental health workers have encountered people with a dual diagnosis in the past, the need to address the problems in the context of community care has never been greater. Literature from the USA suggests that specialist services, which pool resources and skills, are required to provide the specialized support for this client group needs. Existing services in the UK remain specifically focused on substance misuse or mental health, yet the growing body of research indicates the efficacy of providing both services simultaneously, preferably as one discrete service. Finding a pragmatic approach to working with dually diagnosed clients and their families is a challenge to all mental health and drug/alcohol workers and this article describes the efforts of specialist teams to deliver such a service. PMID- 9849156 TI - Transcultural therapy. 1: Mental health and learning disabilities. AB - In this article, the first of two parts, the issues surrounding healthcare provision are discussed in the context of the UK as a multicultural society. Concerns relating to nurse education and training are identified in regard to the implications for practice surrounding diagnosis across cultures, classification of illness, and the status and influence of psychological assessments within the field of mental health and learning disability. Finally, the concept of empowerment is critically reviewed, emphasizing that service providers need to collaborate with communities. Community mental health and learning disability nurses are required to make dynamic alliances with communities, through the individual, and so redirect the power relationship to the community and subsequently respond to the demands for cultural diversity sensitively from within healthcare provision and services. PMID- 9849157 TI - Constipation: cause and control in an acute hospital setting. AB - Constipation is not a glamorous subject, but its prevention and management can make a vast difference to patients' quality of life. This literature review identifies causative factors and describes the mode of action, side-effects and contraindications of many remedies currently available. Recommendations include a systematic approach to management in an acute ward setting, based on assessment of risk, maintenance of normal bowel routine and appropriate dietary advice and the use of appropriate pharmaceutical interventions where necessary. To be effective, this process should be underpinned by accurate documentation and the maintenance of a high level of knowledge and awareness among staff. PMID- 9849158 TI - Cushions and specialist chairs for pressure sore management. AB - Many patients spend more time sitting out of bed than being cared for in bed. Consequently, the demand for specialist support surfaces for chairs to reduce the risk of pressure damage while the patient is up sitting is rising. This product focus examines a number of seating support surfaces produced by Karomed. PMID- 9849159 TI - Proactive risk management: effective communication. AB - Communication is the most powerful tool in clinical practice. Repeatedly, research has shown that good communication skills result in better clinical outcomes, a greater propensity to follow clinical recommendations and reduced risk of clinical negligence and complaints. In using a proactive approach to communication, healthcare professionals must become increasingly sensitized to the stresses associated with illness and hospitalization and must learn the importance of good listening and effective communication to ensure high quality patient care. A study by Lester and Smith (1993) demonstrated that time-limited, negative communications by doctors is associated with increased litigious intentions among patients, even when outcomes were neither adverse nor negligent. The qualities of caring and concern exhibited by doctors make a difference in healthcare outcomes. PMID- 9849161 TI - Nursing urgently needs a professional academy. PMID- 9849160 TI - Ethical dilemmas in clinical supervision. 1: Need for guidelines. AB - In this article, the first of two parts, the authors focus on ethical dilemmas in clinical supervision. Given that there may be several factors that contribute to the limited uptake of clinical supervision, the authors suggest that one reason might be the clinician's concern and confusion regarding the potential ethical dilemmas that could arise when providing clinical supervision. Most issues do not produce ethical dilemmas, but some do. Therefore, this article offers a brief description of the purpose and alleged benefits of supervision, and examines the current position of the law in regard to ethical dilemmas within supervision. It then provides six vignettes where supervisors are faced with ethical dilemmas. These will be discussed in the second part of this article. PMID- 9849162 TI - Who is eligible for the certification exam and why? PMID- 9849163 TI - Vascular birthmarks in children. AB - Several different systems of terminology for vascular birthmarks are defined using descriptive terms (capillary hemangioma, cavernous hemangioma, mixed hemangiomas, etc.) for consistency. Capillary hemangiomas have the most complete and rapid rate of resolution whereas cavernous hemangiomas are considerably slower to respond. Vascular lesions in the V1 distribution of the trigeminal nerve and in the lumbosacral area need imaging studies for presence of internal involvement. Vascular lesions that require systemic therapy are those that obstruct vision, airway, or genitourinary tract; and produce congestive heart failure or consumption of formed blood products. The various modalities of treatment including steroids, are discussed briefly. PMID- 9849164 TI - Handwashing in health care. AB - Noncompliance with handwashing is a significant problem in the health care setting. Numerous studies demonstrate that strategies can be implemented to improve compliance with handwashing. This information has relevance for all areas of nursing practice. PMID- 9849165 TI - Newly recognized infectious exanthems. AB - An exanthem is defined as any rash which is associated with an infectious agent. During the past 6 years, molecular techniques have produced a revolution in clinical dermatology including the exanthems. The infectious agents responsible for many exanthems of previously unknown causes have been uncovered. Included in this review is an update of the rapidly changing spectrum of exanthems, and 10 examples of new viral etiologies of exanthems and two new bacterial exanthems that were recognized during the past 5 years. PMID- 9849166 TI - What's your assessment? Molluscum contagiosum. PMID- 9849167 TI - Health care ethics: models of the provider-patient relationship. AB - Societal changes and ethical issues in health care have been responsible for the evolution of the models of the provider-patient relationship. As health care providers we must recognize the ethical and moral issues inherent in the provider patient relationship and its impact on health care and our society today. PMID- 9849168 TI - Understanding alpha-hydroxy acids. AB - This self-study activity has been developed for nurses, estheticians, and other health care professionals whose patients use alpha-hydroxy acid (AHA) products. Skin anatomy and physiology are reviewed and information is presented on the mechanisms of action of AHAs. Cosmetic and therapeutic benefits of AHA-containing products are detailed. Factors that influence the safety and effectiveness of AHA products are explored, including concentration, pH, and the amount of free acid present. A method is detailed for applying AHA-containing chemexfoliating agents, and guidelines are offered for taking a patient history, post-procedure skin care, and applying subsequent peels. PMID- 9849169 TI - Facial rejuvenation: use of a teaching model in care planning. AB - Rejuvenation of the aging face is in high demand by baby boomers. Facial rejuvenation can consist of various treatments, from chemical peeling with Phenol, TCA, Jessner solution, glycolic acid, and CO2 laser resurfacing. All of these treatments have similar side effects. Nurses play an important role in the educating patients and in treating these side effects. This can be accomplished through a Teaching Model in care Planning. PMID- 9849170 TI - Neonatal dermatology: skin care guidelines. AB - Skin care of the neonate is a very important issue, especially if the infant is either premature or is in critical condition. Guidelines for skin care put special emphasis on the skin barrier properties, transcutaneous absorption, and transepidermal water losses of the skin. The main objective of this article is to give general skin care guidelines for full-term newborn and premature infants, whether in the delivery room, the intensive care unit, or at home. These guidelines are directed to medical staff and nurses with a brief summary for parents. PMID- 9849171 TI - What is your assessment? Perioral dermatitis. PMID- 9849172 TI - Bath PUVA: an effective treatment for psoriasis. AB - At the Psoriasis Education and Research Centre, PUVA baths were introduced as an alternative therapy in 1990. Psoriatic patients soaked for 15 minutes in a solution of tap water and Ultra MOP. Forty-five patients were followed for 30 treatments. Skin types were recorded and progress was documented by PASI (Psoriasis Area and Severity Index) scores. Side effects and total amount of UVA received were documented. The purpose of this study was to record the efficacy of our protocol for PUVA baths on different skin types while evaluating all aspects of the treatment. PMID- 9849173 TI - Exercise guidelines for patients with inflammatory bowel disease. AB - In this article, the author discusses the benefits that people with inflammatory bowel disease derive from a well-rounded exercise program that includes aerobic and resistive training. Exercise guidelines, precautions, and signs and symptoms of potential complications are reviewed. PMID- 9849174 TI - Coping methods of patients with inflammatory bowel disease and prediction of perceived health, functional status, and well-being. AB - Inflammatory bowel disease (IBD) is chronic, incurable, and requires that affected patients cope with their condition on a constant basis. The purpose of this descriptive study was to predict the perceived health, functioning, and well being among patients with IBD through the use and effectiveness of various coping methods. Forty-six patients with IBD completed the Jalowiec Coping Scale, the Health Status Questionnaire, and a demographic questionnaire. Results indicate that use of evasive, fatalistic, and emotive coping methods along with optimistic coping effectiveness were significantly inversely correlated with health perception and well-being. No coping effectiveness variables and only the use of evasive, fatalistic, and supportive coping methods were significantly inversely correlated with perceived functioning. Regression analysis indicated that 20% of the variance in health perception could be predicted from the use of emotive coping method. Fatalistic coping method use explained 24% of the variance in perceived functioning. Finally, 51% of the variability in perceived well-being could be predicted from the use of fatalistic coping method and the effectiveness of optimistic method. These findings indicate that emotion-focused coping methods are important predictors of the health, functioning, and well-being perceptions of patients with IBD. PMID- 9849175 TI - The birth and dissolution of an inflammatory bowel disease support group: lessons in providing support. AB - In response to requests from the members of the local Crohn's and Colitis Foundation chapter, a nurse-facilitated support group was formed. Its goals were to provide support and information for people coping with inflammatory bowel disease (IBD). Despite the interest expressed by the membership, attendance at the monthly meetings was sporadic and sparse. Eventually the group ceased to exist. To understand why the group dissolved, each member was interviewed and asked the following open-ended questions: (1) What did you want from a support group? (2) Why did you not attend meetings regularly? (3) What, if anything, would have kept you coming to the group? Despite coping with IBD, most people described a normal life in which support from the family and established relationships was sufficient to meet their needs. Extra support, such as group support, was only perceived to be needed at certain key times. Innovative methods for providing support, when needed, are discussed. PMID- 9849176 TI - An unexpected outcome after esophagogastroduodenoscopy. PMID- 9849177 TI - Assessment: the key to patient safety when undergoing an endoscopic procedure. PMID- 9849178 TI - Guidelines for preventing allergic reactions to natural rubber latex in the workplace. The Society of Gastroenterology Nurses and Associates, Inc. PMID- 9849179 TI - Increasing oral fluids in chronic constipation in children. AB - Increasing the amount and type of fluid intake in children with simple constipation remains a common intervention recommended by both the medical profession and lay consumers. Efforts to increase overall water intake and/or high osmolarity liquid intake have no research or physiological basis that would result in softer and/or more frequent stools. The purpose of this project was to identify whether an effect on stooling characteristics would be noted with a concerted effort to increase liquid intake. PMID- 9849180 TI - Application of a prototype process for developing a tube gastrostomy clinical pathway. AB - The development and implementation of clinical pathways as a managed care tool have been widely embraced as principal components of the healthcare industry's drive to reform costly, inconsistent, and often uncontrolled delivery of services. Clinical pathways are not new to healthcare; however, the lack of a defined and systematic process for pathway development has hindered organizational efforts to develop and implement clinical pathways. A multidisciplinary clinical pathway process prototype was identified as a means for effectively defining and linking care and outcomes for patients requiring tube gastrostomy placement at a large military medical facility. This article presents the prototype process for developing a clinical pathway with a practical application to illustrate the process and provides a process template for potential use by others interested in developing multidisciplinary clinical pathways. PMID- 9849181 TI - Methemoglobinemia: an unusual complication of topical anesthesia. AB - Allergic reactions tend to be easily recognized and treated, but methemoglobinemia is a rare, life-threatening complication of topical anesthesia of which our staff was unaware until recently. Team work prevented a patient fatality. Directives to alert the staff to suspect methemoglobinemia in patients who develop cyanosis and decreased oxygen saturation levels after administration of topical anesthesia despite administration of oxygen will prevent fatalities in the future. PMID- 9849182 TI - Evaluation and management of the patient with ulcerative colitis. AB - Ulcerative colitis is an inflammatory bowel disease that affects the large intestine. Ulcerative colitis is chronic and can be debilitating. The disease process waxes and wanes, which sometimes gives patients a false sense of cure. The endoscopy laboratory nurse is at the forefront to provide patients with education and offer community support resources. By educating the patient and introducing community outreach programs, the patient can learn to participate and be a partner with the provider in his or her healthcare management. The case study will help outline pertinent information for educating the patient in the management of ulcerative colitis. PMID- 9849183 TI - Medication history and Helicobacter pylori testing. PMID- 9849184 TI - Camp magic: a GI camp. PMID- 9849185 TI - Technology assessment status evaluation: bipolar and multipolar accessories, February 1996. PMID- 9849186 TI - Who is fighting for the right to die? Older women's participation in the Hemlock Society. AB - Who is fighting for the right to die? Past literature has been mixed as to the membership of this social movement. In the current study, 6,398 Hemlock Society members were surveyed in an effort to answer questions concerning who is participating in the right to die movement, whether these participants are rapidly approaching their own death or reacting to the death of a loved one, and whether the movement is invigorated by singular activists. The findings indicate that older, white, wealthy, highly educated, economically and politically active women are in the forefront of the right to die movement. These women report currently being mentally and physically healthy, yet already having taken the steps that will allow them to have an element of control over their death. Finally, right to die support seems to be part of a larger collective network concerning health care and political policy issues. PMID- 9849187 TI - The meaning of home to five elderly women. PMID- 9849188 TI - Older widows' intention to keep the generations separate. AB - As the population ages, increasing numbers of older persons face decisions related to their living arrangements. In predictive studies of coresidence of older parents and their adult children, scholars have not studied older persons' preferred living arrangements. To consider the intentions of 16 older widows with regard to their future situations, a descriptive phenomenological investigation was conducted. The phenomenon "keeping the generations separate" is presented with its two components: continuing to live alone at home and avoiding being a burden to the children. Considered in the contexts of their experience of living alone at home and the life-world, "keeping the generations separate" is a source of insight about influences on older widows' preferences for future living arrangements. PMID- 9849189 TI - Resources and characteristics of elderly women who live alone. AB - This article is a report of findings from a longitudinal descriptive study of 15 women age 76 to 92 who were living alone in the community. Data collection began in 1983. Two survivors were still living alone in 1997. The naturalistic inquiry paradigm guided data collection and analysis (Guba & Lincoln, 1989). Data were collected during repeated audio-taped "conversations" with each woman. Individual "chunks" of data were labeled, sorted, and organized into categories. The 15 categories that evolved from the data were then grouped as resources and characteristics. The prevailing theme was "independence." A woman's value system was a mediator that influenced her ability to continue living alone. PMID- 9849190 TI - Caresharing: hiding frailty in a Florida retirement community. AB - This paper presents research findings generated from a study of the structure of a "caresharing system" for the elderly who reside in a Florida retirement community during the last decade of the twentieth century. A caresharing system is a combination of strategies employed in order to maximize pleasure and minimize losses that might otherwise be associated with communal and individual aging processes. In this instance, the caresharing system entailed a series of conscious efforts to hide frailty in the community. Consequences of such caresharing systems and implications for future retirement communities are discussed. PMID- 9849191 TI - Health, stress, coping, and social support in grandmother caregivers. AB - The purposes of this study were to examine health, stress, coping, and supports of grandmothers who live with one or more grandchildren and participate in their care, and to examine whether differences exist between grandmothers with primary and partial responsibility for their grandchildren's care. A convenience sample of 90 women was recruited from various sites, and participants completed a self administered questionnaire. There were no differences between grandmothers who had primary responsibility for raising their grandchildren (n = 58) and those who did not (n = 32) in self-assessed health, anxiety, or depression; in coping; or in typical grandparenting stresses. Grandmothers with primary responsibility reported significantly greater parenting stresses and less instrumental and subjective social support. Grandmothers in both groups reported high depression and anxiety scores and favorable scores on grandparenting stresses. Implications for grandmother caregivers are discussed. PMID- 9849192 TI - Well-being and its shadow: health promotion implications for older women. AB - In this paper, I describe a theoretical extension to the ecological well-being model of personal, everyday well-being (Ruffing-Rahal, 1989) with the addition of a "shadow model" of core themes and properties. The expanded model enables recognition of transitions in well-being and qualitative experience, thereby providing a venue for deliberate health promotion interventions to address emergent threats to and optimizers of qualitative experience. Illustrative examples, drawn from the investigator's community field experiences involving group health promotion with community-dwelling older women, show the resourcefulness of the shadow model in identifying program topics for timely, relevant preventive and health promotion interventions. The advantages of a small group intervention are also discussed. PMID- 9849193 TI - Female circumcision: persistence amid conflict. PMID- 9849194 TI - The tyranny of feminist methodology in women's health research. AB - Awareness of the insufficient degree to which mainstream research has created useful knowledge about women's health has drawn many researchers to feminist methodologies. Such approaches tend to privilege qualitative designs, emancipatory objectives, and cooperative strategies. They challenge the notions of expert power, the appropriation of voice, and ownership of the research products. By uncovering the extent to which power inequities are embedded in our research traditions, including such issues as who conducts research, which questions are studied, and how they are studied, feminist critique can be a powerful tool toward stronger research with more socially relevant findings. However, taken to extremes, feminist methodological requirements can immobilize and discourage active inquiry. In this paper, we articulate major directives of a feminist stance, explain the extremes at which they become problematic, and propose responsive options for women's health researchers. We intend such analysis to overcome divisiveness and promote inclusiveness without sacrificing excellence in research and action. PMID- 9849195 TI - Abstracting women: essentialism in women's health research. AB - Women's desire to take control of their own bodies creates a natural affinity between the projects of feminism and women's health research. Feminists have used the categories of woman/women, gender, and sex as foundation terms to designate the subject of feminist theories. Universal categories, which have been exposed as essentialist by postmodern and poststructural critiques, create falsely unified subject positions that fail to account for the diversity of women and also fail to acknowledge the situated interests of the dominant groups whose perspectives they reflect. Because it adopts these same categories, research in women's health is also permeated with this essentialized understanding, whether or not it is overtly feminist. In this paper, we point out the dangers of the unreflective use of woman/women, gender, and sex in women's health research. We conclude, that for political purposes, however, a carefully considered "strategic essentialism" can be warranted in research aimed at improving women's health. PMID- 9849196 TI - Conducting feminist research in nursing: personal and political challenges. AB - The challenges of doing feminist nursing research include both personal and political elements. Some of these arise from the threefold influences of being nurses, women, and academics within a larger social context that may be antithetical to feminist values. This paper explores such challenges, using examples from the research of each of the three authors. It includes discussion of such concepts as the tendency to reify certain methodologies and the political forces that may drive research decisions. The authors summarize the challenges of doing feminist nursing research as learning to integrate diverse approaches rather than adhering to a politically correct way of conducting research. They draw on their own research experiences to illustrate the internal conflicts and personal struggles inherent in overcoming the perception that there is one proper way to conduct feminist inquiry. PMID- 9849197 TI - Views of health promotion among primary health care nurses and midwives in Jordan. AB - The purpose of this study was to explore the attitudes of Jordanian nurses and midwives working in primary care settings regarding health promotion. Views of health promotion were examined using a descriptive survey design to measure nurses' perceptions of constraints, responsibilities for health promotion, and client's response to health education. All nurses and midwives (n = 104) working in the Irbid Governorate primary health care (PHC) and maternal and child health care (MCH) centers were surveyed and 95% responded. The data revealed that respondents acknowledged their responsibility to provide health activities and that nurses believed that they were better suited than physicians to do so. Fifty percent of respondents did not believe that lack of time was a barrier to carry out health promotion effectively. However, more midwives than nurses were pessimistic about patient teaching, patient acceptance, and the ability of the provider to make a difference in the patient's health behaviors. PMID- 9849198 TI - Women's reported self-care behaviors during pregnancy. AB - Responsibility for self-care is the purview of the pregnant woman. Her self-care behaviors are influenced by her quest for a healthy pregnancy and by her health care providers and social network. Our purpose in this descriptive ex post facto study was to examine women's reported self-care behaviors during pregnancy. The 100 women in the study sample reported engaging in self-care behaviors that were both positive and negative. More than half of the women reported activities to keep healthy that included walking or jogging, 48 changed their diets, 25 reported working out, exercising, and/or mediating. Of the 32 women experiencing a medical problem with their pregnancies, none reported engaging in behaviors to keep healthy. Assessing reported self-care behaviors early in pregnancy might help identify women who can benefit from advice about self-care strategies to increase their chances of healthy outcomes. PMID- 9849199 TI - From silencing the self to action: experiences of women living with HIV/AIDS. AB - Feminist literature has demonstrated that women often maintain behaviors that support silencing of their voices. The critical issue is whether the silencing experience is (a) a destructive process of burying feelings and needs, (b) a protective strategy to preserve personal and professional relationships which they value, (c) a coping mechanism to divorce themselves from an androcentric/ethnocentric health care culture, or all of these. The transition from silence to action may be a process of reacting to a threat to self (i.e., HIV/AIDS diagnosis) where gender normative behaviors become irrelevant and self advocacy becomes paramount for survival. Alternatively, the transition may be a conscious process of gaining insight into past behaviors that have been learned and culturally supported and making purposeful changes. Data for this study were extracted for secondary analysis from data from a larger study on experiences and needs of persons living with HIV/AIDS. Data were obtained from transcripts from three focus groups (N = 14 women) and six individual interviews. Women ranged in age from 21 to 55; 9 were European American, 7 were African American, and 4 were Latina American. Data were content analyzed and organized using four categories proposed by Jack (1991): (a) externalized self-perception, (b) care as self sacrifice, (c) silencing the self, and (d) the divided self. Data supported that women with HIV/AIDS reported all four categories of silencing behaviors, particularly early in the HIV trajectory. For some women, an HIV/AIDS diagnosis ignited them to speak for themselves and to shape their own lives based on feelings and needs. For others, peer or professional support or both was the catalyst for the transition from silence to action. Findings suggest interventions that would assist women in judging themselves by internal versus external standards, putting their own needs before the perceived needs of others, expressing themselves toward action rather than avoidance, and feeling comfortable expressing anger. PMID- 9849200 TI - A correlational framework for understanding sexuality in women age 50 and older. AB - To investigate a biopsychosocial perspective of sexuality in women age 50 and older, 657 community-residing women completed anonymous surveys, including demographics; health; self-esteem; intimacy; and sexual knowledge, attitudes, interest, participation, and satisfaction. Cronbach's alphas for major variables ranged from .52 (sexual knowledge) to .92 (intimacy). Mean scores for major variables ranged from 2.96 (sexual participation) to 3.43 (self-esteem, sexual attitudes). Significant correlations were described among the predictor variables and sexual interest, participation, and satisfaction. Significant predictors of sexual interest, participation, and satisfaction were identified through stepwise multiple regression procedures. Findings suggest a biopsychosocial model for clinicians to use in assessment and intervention with older women and sexual issues, for educators to use to organize teaching about aging and female sexuality, and for researchers to use to investigate older women's sexuality in different samples of older women. PMID- 9849201 TI - In search of childbirth knowledge. AB - My purpose in writing this article is to describe the various ways expectant women acquire childbirth knowledge. The data presented here form a small section of a bigger study that examined how expectant women from two cultures prepare for childbirth. The aims of the study were to explore the reasons why some expectant women choose to attend preparation classes run by non-health-care professionals and to describe the ways childbirth knowledge was disseminated to women at the preparation classes. Interviews containing structured and open-ended questions were used to obtain data. The first interview followed completion of a series of eight preparation classes and the second was conducted one month after delivery. Observation techniques were used to collect data on the teaching strategies and the content of classes. Ninety-five expectant women participated in the study. Data analysis included frequency counts and percentages on all structured questions. The open-ended questions were analyzed using the constant comparative method to identify major categories and themes. These were reviewed by a panel of childbirth educators for reliability. Frequencies were calculated for the number of responses in each category. Findings show that women acquire childbirth knowledge in various ways at the classes. They used this information to negotiate their care with health care professionals. PMID- 9849202 TI - Women's health and community health nursing practice in geographically isolated settings: a Canadian perspective. AB - In this article we examine the literature on women's health and community health nursing practice in geographically isolated settings from a Canadian perspective. An extensive review of journal literature, Canadian government documents and reports, and reports by women's organizations forms the basis of this examination. Following a brief introduction to the concept of geographical isolation, literature relating to women's health issues in rural and northern settings is described. This is followed by a discussion of community health nursing practice and women's health in rural and northern settings. The discussion section identifies major gaps in the literature and suggests implications for community health nursing practice and research, as well as strategies for policy and programming to improve women's health and community health nursing practice in geographically isolated settings in Canada. PMID- 9849203 TI - Looking to care or caring to look? Technology and the rise of spectacular nursing. AB - New technologies, such as ultrasonography and vital function monitoring, have made nursing care more spectacular. Nurses now watch, in addition to watching over, their patients. Nursing practice increasingly shapes and is shaped by simulations and representations of the real. Visualist technologies represent a new way of knowing in nursing and a new area of nursing informatics. They also raise all the esthetic, political, and ethical dilemmas associated with looking. They raise the question: Do nurses look to care, or care to look? PMID- 9849205 TI - The coalescence of technology and humanism in nursing practice: it doesn't just happen and it doesn't come easily. AB - Health care technology occurs in the cultural and organizational context of users and providers and thus is inherently neither "good" or "bad." Situated at the midpoint of the technologic-humanistic dualism, registered nurses facilitate the coalescence of technology and humanism in nursing practice. This coalescence does not just happen and does not come easily. It requires both caring and expertise and is facilitated by education, clinical practice, research, and administrative considerations. This coalescence may indeed lead to some of the most exciting breakthroughs of the 21st century in terms of what it means to be human. PMID- 9849204 TI - Who really makes the bed? Uncovering technologic dissonance in nursing. AB - Technologic dissonance in the practice arena is demonstrated in the use of nonnursing technologies that are present in but technically incongruent with a nursing environment. Technology in nursing is not necessarily nursing technology. The article traces the influences of technologic dissonance beyond the reductionist purview of the medical model to their source in design and engineering philosophies. The vision of technologic consonance in nursing may be realized in the influence and expression of the caring nurse in the design, engineering, ownership, and use of nursing technology. PMID- 9849206 TI - Discovering the value of nursing in high-technology environments: outcomes revisited. AB - The article argues that the current approach to specifying outcomes of nursing care in high-technology environments obscures rather than illuminates nursing's contributions to health care. Two nursing situations are used to illustrate a reframing of the idea of outcomes from the perspective and language of nursing as caring. Outcomes of nursing care are reconceptualized as value experienced within the nursing situation. PMID- 9849207 TI - Technology and true presence in nursing. AB - If there is to be a future for nursing in the next millenium, it must be dictated not by technologic advances directly but by the manner in which technology relates with persons in health and illness. Technology enables nurses to care for persons in a more efficient and cost-effective manner. True presence, as articulated in the human becoming theory, affords members of the profession opportunities to be with persons in ways that make a difference to their quality of life. The article illustrates the recognition of the link between technology and true presence through the use of a nurse-person encounter. PMID- 9849208 TI - Technologic competence as caring in critical care nursing. AB - The article describes technologic competence as caring in critical care nursing, a framework grounded in the perspective of nursing as caring. The achievement of technologic competence is an exercise in the process of knowing the wholeness of persons in the moment. Nurturing persons toward well-being is the focus of nursing. Technology, caring, and competence are core concepts that constitute the framework of technologic competence as caring in critical care nursing. Technologic competence epitomizes critical care nursing and assumes an indispensable position in contemporary nursing practice. PMID- 9849209 TI - Technology and care in a bone marrow transplant unit: creating and assuaging vulnerability. AB - The article describes the experience of technologically induced vulnerability and the inherent uncertainty of patients undergoing bone marrow transplantation. Examples of care by nurses, as perceived by patients and their family members, are offered. The relationship between the iatrogenic vulnerability and suffering of patients and the nursing response of care is explored. The claim is made that a caring response by nurses enables patients to make meaning of their choice to undergo simultaneously life-saving and life-threatening bone marrow transplantation. PMID- 9849210 TI - A phenomenologic study of the interface of caring and technology in intermediate care: toward a reflexive ethics for clinical practice. AB - A phenomenologic research study of nurse caring was conducted in an intermediate care unit of a hospital. Three narratives from the data are presented as examples to illuminate the meaning of the experience of caring for technologically dependent patients. Results revealed that vulnerability, suffering, and the ethical situations of moral blurring and moral blindness were the dynamics of caring for these patients. Identification of these phenomena cells for a new reflexive ethics in health care to increase understanding of how deep values in relation to the status of vulnerable human beings, and how moral interactions among patients, families, and caregivers, shape ethical decisions. PMID- 9849211 TI - The meaning of technology for people with chronic renal failure. AB - The article is based on the findings of a hermeneutic study that focused on the experience of individuals receiving hemodialysis for chronic renal failure. The purpose of the research was to begin to understand the experience and meaning of technology for these individuals. Three themes reflected the dimensions of the participants' individual and collective experiences: coming to terms with loss and limitations, abiding with technology, and enduring the treatment environment. Overall, the participants found themselves struggling to remain embodied while being transformed by a reluctant partnering with technology. PMID- 9849212 TI - Melody link. PMID- 9849213 TI - When it comes to advance directives, we can't afford question marks. PMID- 9849214 TI - Moments of courage reconciling the real and ideal in the clinical practicum. PMID- 9849215 TI - Maintaining patient confidentiality in today's health care environment. PMID- 9849216 TI - Nursing and ethical issues. PMID- 9849217 TI - Student academic dishonesty. PMID- 9849218 TI - When your best may not be good enough. PMID- 9849219 TI - Jeffrey Wright. Interview by Molly Berkowitz. PMID- 9849220 TI - Domestic violence today. What nursing students should know. PMID- 9849221 TI - The Open Window Phase offers hope for battered women. PMID- 9849222 TI - It happened to me. PMID- 9849223 TI - Imprint special section travel nursing. PMID- 9849225 TI - Up close. Kristin Helland. Interview by Kimberly Allen. PMID- 9849224 TI - Four days at an AIDS hospice. PMID- 9849226 TI - Making a difference in smoking cessation interventions. PMID- 9849227 TI - The importance of resolutions in influencing policy. PMID- 9849229 TI - Juggling nursing school and family. A snowstorm and a missing shoe take their toll. PMID- 9849228 TI - Embracing professionalism through NSNA. The Grand View College experience. PMID- 9849230 TI - How to love nursing theory. PMID- 9849231 TI - Personal budgeting for nursing students. The care plan for a healthy checkbook. PMID- 9849232 TI - Resource books for nursing students. PMID- 9849233 TI - Now--the history of the future. PMID- 9849234 TI - Determining patients' suitability for thrombolysis: coronary care nurses' agreement with an expert cardiological 'gold standard' as assessed by clinical and electrocardiographic 'vignettes'. AB - BACKGROUND: Thrombolytic therapy is an established treatment for acute myocardial infarction, the benefits of which are time-dependent. Coronary care nurses are often the first professionals to assess patients with suspected myocardial infarction on hospital admission, and there are a number of reports of nurseled practices for immediate assessment and 'cardiac triage'. OBJECTIVE: To assess coronary care nurses' ability to determine patients' suitability for thrombolysis as judged by an expert cardiological 'gold standard'. METHODS: Ten coronary care units in the Northern and Yorkshire region of England were sent packs comprising 10 sets of 10 'vignettes' and accompanying electrocardiograms together with a questionnaire regarding treatment decisions. An expert 'gold standard' for correct responses to the questionnaire was derived following independent assessment of the vignettes by three nurse specialists and three consultant cardiologists. Respondents' treatment decisions were subsequently compared with the 'gold standard'. RESULTS: Of 42 (42%) completed sets of questionnaires returned for analysis, 37 (87%) of respondents indicated management decisions in absolute agreement with the 'gold standard'. Where there was not absolute agreement, most treatment decisions were acceptable as safe practice. CONCLUSION: The majority of nurses indicated safe and appropriate management decisions. These limited data provide further support for ongoing developments of nurse-initiated thrombolysis, although there is a need for further, larger trials to determine the safety and efficacy of such an approach to patient care. PMID- 9849235 TI - Intensive nursing care of patients with a microvascular free flap after maxillofacial surgery. AB - This article provides an introduction to the care of patients following maxillofacial surgery, many of whom undergo the complexities of microvascular flap surgery and need careful nursing assessment in the postoperative period. A brief introduction to this surgery illustrates some of the potential reasons for admission to the intensive care unit. The nursing care is vital to maintain the survival of the flap, the details of which are discussed along with factors which potentially contribute to flap failure. Other considerations such as haemodynamic stability, postoperative complications, the importance of education and other options for postoperative management also form further points for discussion. PMID- 9849236 TI - Barriers to evidence-based practice. AB - OBJECTIVES: This paper reports the findings of a study which aimed to identify the barriers to evidence-based practice in an acute National Health Service (NHS) trust. The study was carried out as part of an action research project designed to promote evidence-based practice. DESIGN: A rapid organizational appraisal design was used. This involved formal and informal interviews, focus groups, and observation of staff interactions at meetings and in clinical practice in an acute hospital NHS trust. Interviews were undertaken with key stakeholders within the organization and focus groups held with nurse managers, audit staff, staff nurses, and junior medical staff. Observation took place in meetings and in clinical areas. RESULTS: At an organizational level, the main issues identified were: (1) evidence-based practice was a low management priority, (2) problems with dissemination, (3) inadequate systems for personal and professional development, (4) difficulties in the management of innovations, (5) accessing evidence and resource constraints. At the individual practice level, the main issues were motivation, a lack of clarity about roles and practice, and a culture of practice which emphasizes 'routine' patient care. CONCLUSIONS: The results of this study demonstrate that structures and cultures within organizations can be important barriers to evidence-based practice. Factors which are external to individual trusts are also important. Existing hierarchical structures both in the NHS and within and between the different professional groups, are manifest in the existence of a largely deferential culture which emphasizes the routine in practice decision making. Given this reality, organizations will have to adopt multiple strategies to facilitate and promote the use of evidence in practice decision making. PMID- 9849237 TI - To bag or not to bag? Manual hyperinflation in intensive care. AB - Manual hyperinflation or bagging is a physiotherapy technique commonly used on mechanically ventilated patients on intensive care. The way it is performed appears to vary widely, and in the past there has been little or no monitoring of what is actually happening during the technique in terms of what airway pressures and tidal volumes are being delivered to the patient. Conclusive evidence from large scientific studies of the benefits and side effects of manual hyperinflation are to date still lacking. PMID- 9849238 TI - The principles of augmenting the cardiac output in adults. AB - Augmentation of the patient's cardiac output to optimize tissue oxygenation is a primary role of intensive and cardiac care nurses. This involves three approaches: the use of intravenous colloids and occasionally crystalloids, creating a passive stretching of the myocardium to increase cardiac output, the use of inotropic agents which directly stimulate the heart and finally and less commonly the use of cardiac assist devices, notably the intra aortic balloon pump. The choice of method will be dictated by the presenting clinical data and the perceived benefits to the patient of each approach. The role of nurses as part of the health care team is to practice safe, informed care, through an understanding of both the advantages and disadvantages of each approach to the clinical scenario encountered. This paper contains exploration of each of the three principles with a case study approach, allowing the reader to consider a variety of presenting clinical data and appropriate haemodynamic manipulation. PMID- 9849239 TI - The evolution and nursing history of a general intensive care unit (1962-1983). AB - In this paper we have reviewed the origin and evolution of Whiston Hospital's General Intensive Care Unit (ICU) from its humble beginnings as an offshoot of a general ward in the early 1960s. The length of service of the senior nursing staff over a period of 21 years was also calculated. The average duration of service was 16.5 years--a figure which significantly surpassed those quoted in the literature. In addition, we have outlined the development of nurse training in intensive care as well as the role of the nurses in research in the ICU. The question as to whether later college-based training was superior to the previous hospital-based course remained unanswered. PMID- 9849240 TI - Analgesics in the management of chronic pain. Part Three: Step 2 analgesic drug therapy. AB - In this, the third article in the series, the role of analgesic drugs appropriate for the treatment of moderate to severe pain is reviewed along with therapies frequently applied in the treatment of poorly responsive neurogenic pain. The article should be read in conjunction with part one outlining general principles in the management of chronic pain. PMID- 9849241 TI - Clinical effectiveness: an old ideal but with a newer context and approach in nursing. PMID- 9849242 TI - Contextualizing critical care family needs through triangulation: an Australian study. AB - Family needs and concerns within the critical care context have been thoroughly explored from the quantitative perspective utilizing the Critical Care Family Needs Inventory (CCFNI). Nursing interventions have been designed on the basis of the findings from these studies. However, while the CCFNI would seem to encompass all the possible needs of families with a critically ill loved one, at no time were the family members themselves consulted at length in regard to the development of the instrument, or in any subsequent validation studies. Individual reality generates the variables that are measured in a needs analysis, and the family member experiences encompass dimensions that are not easily assessed by quantification. In fact the unique experiences of family members underpin their perception of need. Methodological triangulation formed the basis for this study to determine the degree of confirmation (or otherwise) between family member respondents to the CCFNI (n = 105) and those participating in an interview (n = 26) designed to explore needs and experiences. The qualitative data served the purpose of completeness by providing a more contextual representation of needs and therefore greater depth of understanding of the whole construct. The results indicate that, while there were many areas of convergence between the two samples, there were also areas of diveregence. Two major needs emerged from the interviews that are not represented on the CCFNI: the need of family members to provide reassurance and support to the patient; and their need to protect (others as well as the patient). A more complete understanding of family needs was obtained through the contextualization of their experiences. PMID- 9849243 TI - Can nurses safely assess the need for endotracheal suction in short-term ventilated patients, instead of using routine techniques? AB - Most literature describes endotracheal suction as a hazardous procedure associated with numerous complications and proposes that it should only be performed as necessary to minimize these complications. Other authors suggest endotracheal suction only after assessment predisposes patients to a number of different complications. This article describes a controlled study to compare and contrast the differences in endotracheal suction outcomes in patients who received ritualized 2 hourly suctioning and those who received it following assessment. A group of qualified nurses in an Intensive Care Unit were taught auscultation skills to assess a patient's needs for suctioning and all the nurses received educational training regarding endotracheal suctioning. Short-term ventilated patients were allocated to receive endotracheal suctioning either when the need for it was determined by assessment only or routinely, using a standardized suctioning technique. The results demonstrated a clear increase in nurses' knowledge regarding endotracheal suctioning. The assessed group of patients demonstrated significantly better outcomes and less complications than the controlled group in relation to changes in peak airway pressures, heart rate and mean arterial pressure pre- and post-endotracheal suctioning, and the amount of secretions obtained on suctioning. Although only preliminary, these results do provide support for the view that endotracheal suction only in response to assessment is better practice for short-term ventilated patients. PMID- 9849244 TI - The role of advocacy in critical care nursing: a caring response to another. AB - With the aim of clarifying critical care nursing, six critical care nurses, working in a Swedish intensive care unit (ICU) were each asked to narrate a care situation with which they had been satisfied or dissatisfied. The stories were tape-recorded and analysed using a phenomenological-hermeneutic approach inspired by the philosophy of Ricoeur. The themes uncovered in the analysis describe the nurse's role of advocacy as: to build a caring relationship, to carry out a commitment, to empower, to make room for and interconnect, to be a risk-taker, to be a moral agent and to create a trusting atmosphere conducive to recovery. The meaning of the role of advocacy lies in a moral and existential response to another human being, an expression of caring. Advocacy rests on the patient-nurse relationship and occurs as an outspoken demand of another human being whose autonomy is threatened. The results are discussed from the ethical perspectives of Logstrup, Watson's concept of care, and existential advocacy as expressed by Gadow. PMID- 9849245 TI - A randomized trial comparing Arglaes (a transparent dressing containing silver ions) to Tegaderm (a transparent polyurethane dressing) for dressing peripheral arterial catheters and central vascular catheters. AB - The purpose of this trial was to prepare for a large randomized trial comparing Arglaes film dressing, a recent innovation containing silver ions, against Tegaderm, a transparent polyurethane dressing. Thirty-one patients admitted to the intensive care unit and requiring the insertion of an arterial line or central venous catheter were recruited into the study. Skin swabs were taken from the insertion sites prior to catheterization and on removal of the intravascular device to measure skin colonization rate between the two dressings. The catheter tips were also cultured on removal to establish if there was a difference between the two groups. No statistical differences were found in bacterial growth between the two dressings. PMID- 9849246 TI - A comparative study of differences in the referral behaviour patterns of men and women who have experienced cardiac-related chest pain. AB - Survival and long-term prognosis after a myocardial infarction are directly related to the individuals's decision to seek medical help. Early medical intervention is imperative if thrombolytic therapy is to be effective. Emerging research has indicated that women frequently have longer pre-hospital delays than men. Incorporating a comparative descriptive design, this research compared the pre-hospital admission behaviour patterns of women and men, following the onset of acute chest pain. A convenience sample of 12 women and 12 men were selected following admission to a local coronary care unit. Structured interviews and the examination of medical notes/relevant documentation provided the data. Findings were examined and compared through the use of content analysis and descriptive statistics. Severity of symptoms proved to be the strongest influence in shortening pre-hospital delay. Despite this, overall, men were admitted to hospital more quickly than women. Men were more ready than women to believe that they might be having a heart attack and this belief led them to seek treatment promptly. In order to improve female mortality and morbidity following a myocardial infarction, it is recommended that there is an urgent need to target women, through health promotion and media interventions. PMID- 9849247 TI - Family nursing in intensive care. Part two: The needs of a family with a member in intensive care. AB - It was demonstrated in Part one that the family is a justifiable concern to nurses in intensive care and that family-focused care is appropriate in such an area. If this approach to care is to be considered, it is necessary to identify the needs of families when they have a member in an intensive care unit. This is a well-researched area and some of the relevant literature is discussed within this paper. Before this literature is addressed, an attempt is made to define the concept of need. The methodologies used in the identification of family needs could be adapted to explore whether or not the needs of the families of patients in a particular intensive care unit are being met. This paper suggests that if it was shown that the meeting of family needs was an area of care which could be improved upon, then a family-centred approach to care would be a reasonable option. If however, it is shown that families already perceived their needs as being met, then changing the system of care to a more formal family nursing approach would seem to be unnecessary. PMID- 9849248 TI - Is a murder charge an occupational hazard of intensive care nursing? AB - When nurses are accused of harming their patients there is an understandable wave of concern. When nurses kill their patients there is disbelief and horror. After all, killing patients is so far from the traditional image of the nurse as to cause distress and disorientation. When the nurse in question is a woman and when the charge amounts to serial killing, our most cherished assumptions about gender roles and professional responsibility are called into question. Yet in this decade nurses have been accused of killing patients, attempting to kill patients and causing grievous bodily harm to others. In the UK, of the nurses accused of attempting to murder their patients two came from the same specialism. Two of the three accusations centred on Intensive Care Units (ICU); the case of Amanda Jenkinson (Kenny 1996) and the case of Kath Atkinson, an ICU sister in Newcastle (Porter 1998a). In the most notorious case of nurse homicide in the UK this decade, that of paediatric nurse Beverly Allitt, the profession and society at large were shaken by accusations so serious as to defy belief. PMID- 9849249 TI - Analgesics in the management of chronic pain. Part two: Step I analgesic drug therapy. AB - This, the second in a series of five articles, reviews the role of simple analgesics and co-analgesia in the treatment of chronic pain. It should be read in conjunction with Part one which covers general principles of pain management. PMID- 9849250 TI - To tell you the truth: children reflect on hospital care. AB - Children enter hospitals as novices and leave them as social critics. Their "take" on the environment and their reaction to care received are outcome measures too often neglected in our fast-paced health care arena. This article publishes the evaluations of these young patients to an audience of health professionals ... practitioners who might be moved by such poignant expressions of thanks and motivated by such thoughtful prescriptions for reform. Much of the data were gathered during the development of the Web site, "Band-Aides and Blackboards: When Chronic Illness ... Or Some Other Medical Problem ... Goes to School." PMID- 9849251 TI - Use of a family caregiving model to articulate the role of the public health nurse in infant mental health promotion. AB - Public health nurses (PHNs) have dedicated their efforts to promoting the health and welfare of mothers, infants, and children, receiving little recognition for the mental health promotion benefits they provide to these populations. The purpose of this article is to describe the contributions of PHNs to the promotion of mental health in infants and their families. The Family Caregiving Model (Zerwekh, 1991) is used to identify the strategies used by PHNs to promote infant mental health within vulnerable families. Using a case study format, the authors describe these strategies and the competencies and skills needed by PHNs to assess for cues in infants and their parents that may indicate the potential for unsuccessful attachments. PMID- 9849252 TI - Preschool children of battered women identified in a community setting. AB - This study describes and compares health, developmental, and behavioral manifestations of witnessing domestic violence in a health department population of preschool children (n = 15) of women detected to be battered (Conflict Tactics Scale [CTS], Straus, 1979). Comparisons included a community sample of the preschool children of nonbattered women (n = 62) to study the effect of mothers being battered, and a shelter sample of the preschool children of battered women (n = 53) to study the effect of "sheltering" on children's characteristics. Instrumentation included behavioral subscales, physical assessment, Denver II Developmental Screening, hemoglobin and lead levels, immunization, and nonmotor vehicle accident histories. Results showed more similarities between children of community (nonsheltered), battered and nonbattered women, but showed more differences between the children of community (nonsheltered) battered women and the children of sheltered, battered women. Battering intensity of the women was more severe in the shelter sample according to the CTS (p < .001), and more abnormal findings in their children were detected than in the community sample. When children of community (nonsheltered), battered women (n = 15) and children of sheltered, battered women (n = 53) were included in discriminant analysis of demographic characteristics, health status, and psychosocial variables, 87% of each group was classified correctly. PMID- 9849253 TI - The contribution of health promotion to community children's nursing. AB - This article discusses the range of current thinking on health promotion with special reference to community children's nursing. The scene is set with the identification of a number of issues currently at the heart of the promotion of health in children and young people. Emphasis is placed on the unprecedented expansion of interest in the promotion of children's health in Northern Ireland. Current meanings attributed to health promotion are discussed and applied to the practice of community children's nursing. Some of the contentious elements of these definitions and meanings are debated. For example, the distinct meanings of health and well-being in children are discussed and the issue of whether health promotion should be aimed at individual children, their families, or their communities is pondered. Prevention, it is argued, is one particularly important component of the promotion of health in children in the community in Northern Ireland. PMID- 9849254 TI - Are persons multidimensional or holistic? PMID- 9849255 TI - Immigration and ethnicity. Implications for holistic nursing. AB - International labor migration has increased the day-to-day encounters of persons from different cultural groups. The concept of ethnicity, its historical development, its ambiguity, and its role in the interactions between persons of different cultural groups are explored. The arena of health care for migrants brings to the fore issues of providing culturally competent care. In an ideal world, all nursing care of migrants would be delivered by skilled transcultural nurses; but in the real world, this is not yet the case. A case study of a Mexican migrant woman and a non-Hispanic Midwestern nurse is used as a background for examining the role of ethnicity in determining care needs and expectations and for providing nurses with a perspective that can improve nurse-client collaboration. PMID- 9849256 TI - "Codependency". A disease or the root of nursing excellence? AB - A hermeneutic phenomenological study was conducted to explore how eight professionally competent nurses experienced and evaluated the relation between their childhood adaptation to dysfunctional families and their nursing careers. From the participants' discussion of this topic, the following themes emerged: escaping difficulties by becoming a nurse, coping roles guide nursing career, sensitivity to the untold, transforming dysfunctional responses, and wounded healers. The study did not support the view that children of alcoholics seek careers in nursing to meet their codependent needs for self-esteem, control, or belonging. Instead, its findings indicate that some children of alcoholics become competent nurses by finding positive application for the coping skills they learn in their families. This indicates that, when working with individuals from dysfunctional families, nurses could support them to create new avenues for their coping skills instead of trying to "exterminate" them because of their "codependent" nature. PMID- 9849257 TI - Spiritual nursing interventions. AB - Oncology, parish, and hospice nurses in the Midwest were surveyed to explore what nursing interventions they implement to enhance the spirituality of clients and how they learned about these interventions. Some 95 spiritual nursing interventions were identified and ranked according to frequency of implementation. The most frequently identified nursing interventions were referral, prayer, active listening, facilitation and validation of clients' feelings and thoughts, conveying acceptance, and instilling hope. The most frequently ranked implemented interventions were communication and religious nursing interventions. Only 15% of the participants indicated that they learned the most about spirituality and spiritual nursing interventions from basic and advanced nursing education combined. Findings support the need for increased emphasis of theoretical and clinical spiritual knowledge in nursing education and the development of a typology of spiritual nursing interventions to direct nurses in enhancing the spirituality of clients and implementing holistic nursing care. PMID- 9849258 TI - Parish nursing. Addressing the significance of social support and spirituality for sustained health-promoting behaviors in the elderly. AB - Literature about the significance of health promotion for the elderly and identification of various determinants of health-promoting activities contains numerous inconsistencies. From a review of nursing and health-related literature over the past decade, it is possible to draw only limited conclusions about the commitment of the elderly to health-promoting behaviors. Despite this, the importance of the roles of social support and spirituality for the empowerment of older adults to practice health-promoting activities is a consistent research finding in this population. Parish nurse programs expand home health and public health provider roles and use the faith community as a cooperative means for sociocultural implementation of successful health promotion. Significance for nursing education, practice, and research is discussed. PMID- 9849259 TI - Pain beliefs. What do elders believe? AB - The purpose of this descriptive study was to better understand elders' beliefs regarding chronic pain. The research questions were (a) What are the pain beliefs of elders experiencing nonmalignant chronic pain (NMP) and chronic pain associated with malignancy with the hope for recovery (MHR)? (b) Are there differences in pain beliefs between the NMP group and the MHR group? and (c) Do elders have similar or different pain beliefs? Sixty adults age 65 or older, all cognitively intact, completed the Pain Beliefs Questionnaire; 30 had arthritis and 30 had cancer. Of the total, 60% responded that pain is the result of damage to the body--either often, almost always, or always. Another 13% believed that pain was rarely or never the result of such damage, and 15% rarely or never saw pain as a sign of illness. In all, 68% responded that depression makes pain seem worse. PMID- 9849260 TI - Effect of therapeutic touch on the well-being of persons with terminal cancer. AB - The purpose of this study was to examine the effect of three Therapeutic Touch treatments on the well-being of 20 persons with terminal cancer in palliative care. Participants in the experimental group (n = 10) received three noncontact Therapeutic Touch treatments, the duration of which varied between 15 to 20 minutes. Participants in the control group (n = 10) participated in three rest periods. Well-being was measured at preintervention time and immediately postintervention time using the Well-Being Scale, a visual analogue scale measuring pain, nausea, depression, anxiety, shortness of breath, activity, appetite, relaxation, and inner peace. The results of the study support the hypothesis that three noncontact Therapeutic Touch treatments increase sensation of well-being in persons with terminal cancer. PMID- 9849261 TI - Faculty-student mentoring. A father's chronic sorrow: a daughter's perspective. AB - Chronic sorrow often affects not only an individual who has a chronic illness or disability but the family members as well. This is the story of how chronic sorrow affected the life of a man diagnosed with prostate cancer and the family members who lived through it with him. It is told in the first person by his daughter not only as a tribute to her father, but also as a prospective learning experience for anyone involved with an individual moving through a chronic or terminal illness or disability. It integrates theory, research, and personal experience in an effort to understand this common human response to a chronic or terminal illness. PMID- 9849262 TI - Finding the "dad" in unwed fathers. PMID- 9849263 TI - Infectious diseases and the immunizations of today and tomorrow. AB - For decades, health care providers have administered immunizations for common childhood infections. These have been given at predicted times according to an agreed-upon schedule. However, in the past decade, common immunizations have been changed, the number of immunizations has increased, and the schedule for administration of vaccines officially changes every January. In addition, as science continues to grow, new vaccines are being developed faster than ever before. PMID- 9849264 TI - Rotavirus: an overview--from discovery to vaccine. AB - Rotavirus is the single most important cause of both nosocomially-acquired and severe, dehydrating diarrhea. It occurs with equal frequency throughout the world, regardless of the level of sanitation. After primary infection, mild or asymptomatic reinfection is common. Current management involves the use of oral rehydration therapy and early feeding of an age-appropriate diet. Cereal-based oral rehydration solutions, lactic acid bacteria, and oral immunoglobulins may play a role in future treatment. A rhesus rotavirus, quadravalent oral vaccine, currently being considered for licensure, may be given concurrently with other routine infant immunizations and has an efficacy rate of 80% against severe disease. Pediatric nurses should be aware of these recent advances in treatment and prevention, which should significantly reduce the impact of rotavirus in the near future. PMID- 9849265 TI - Human parvovirus B19--flushed in face though healthy (fifth disease and more). AB - Parvovirus B19 is an ubiquitous organism. Although for many years it had been "a virus in search of a disease," it is now known to be associated with several clinical entities ranging from the benign to severe. The most significant aspect of parvovirus B19 is its effect on the fetus. Research has demonstrated the risk, although minimal, of fetal loss to be between 1.5% to 2.5%. The effect from an emotional standpoint for the pregnant exposed or infected woman is more difficult to quantify. It is imperative that nurses who care for children be well informed about the virus and able to implement a comprehensive plan. PMID- 9849266 TI - Operationalizing Donna Wong's principle of atraumatic care: pain management protocol in the NICU. AB - Atraumatic care is the philosophy of providing therapeutic care through the use of interventions that eliminate or minimize the psychologic and physical distress experienced by children and families (Whaley & Wong, 1995). The foundation of this principle lies in minimizing separation of child from family, identifying child/family stressors, minimizing/preventing pain, and promoting parent professional partnerships. A review of pain management practices in one neonatal intensive care unit demonstrated underprescription and under-administration of pain medication as well as inconsistencies among practitioners. Written guidelines, developed by an interdisciplinary team and provided here, integrated current research in the assessment and management of procedural, postoperative, and other pain in infants. Use of the pain management guidelines, which serve as a foundation for the unit's commitment to atraumatic care, have improved pain management practices and have standardized care for infants on the unit. PMID- 9849267 TI - Long-term follow-up of survivors of neonatal ECMO: what do we really know? AB - Extracorporeal membrane oxygenation (ECMO) has been used increasingly since the 1970s to treat serious respiratory diseases of infancy. However, much is unknown regarding developmental processes and outcomes in these high-risk children. Since the majority of children treated with ECMO have "normal" intelligence, they are often deemed developmentally appropriate by families and primary care providers in early childhood. Unfortunately, many children may exhibit cognitive deficits later, falling behind their peers in school during middle childhood. Predicted academic success cannot be based on IQ alone. Studies have shown that between 10% and 35% of ECMO survivors who are developing "normally" are in need of special educational services due to attention problems and/or express variability in functional abilities. Although the research data remain inconclusive, parents of children who undergo neonatal ECMO should be alerted to potential problems so that children can receive full assessments and appropriate interventions. PMID- 9849268 TI - Preventing lead poisoning and its consequences. AB - The harmful effects of lead poisoning have been clearly espoused in the literature. All health care providers should be aware of these dangers and of current recommendations for screening for lead poisoning risks and follow-up of blood lead levels. Health education is an important aspect of health care. Pediatric nurses need to understand the hazards of lead poisoning, screening and follow-up recommendations, and have information about lead poisoning and abatement procedures available to families. PMID- 9849269 TI - Pediatric management problems. Left corneal abrasion. PMID- 9849270 TI - Vigabatrin. PMID- 9849271 TI - Stress-point intervention for parents of children hospitalized with chronic conditions. AB - Stress-point intervention by Nurses (SPIN) is a research-based intervention that can be used to assist families whose child has a chronic condition and is repeatedly hospitalized. The intervention addresses attendant family stressors, tasks, and concerns. SPIN incorporates parent-nurse contact before admission, during hospitalization, and after discharge. A nurse assists each family to develop their unique set of coping strategies, which are shaped by the family's own concerns and resources and the nurse's expertise. The effectiveness of SPIN has been tested in two intervention studies, one based in the community and another based in ambulatory clinics. Both studies found that families receiving SPIN interventions had better parental coping and family functioning, and there was less developmental regression in the child after hospitalization. SPIN can be used by nurses having experience working with families and children. A self directed learning program assists with understanding of theoretical and practical aspects of SPIN. PMID- 9849272 TI - Improving ventilation in children using bilevel positive airway pressure. AB - Noninvasive positive pressure ventilation by way of a mask, commonly known as BiPAP, has become a widely used procedure to support patients with respiratory failure, both in the chronic and the acute settings. Currently, this mode of ventilation has been extended to the pediatric population. This article focuses on the use of BiPAP in pediatric patients. Its purpose, potential situations for use, mode of functioning, and nursing implications will be discussed. PMID- 9849273 TI - Infant physiological responses to noxious stimuli of circumcision with anesthesia and analgesia. AB - PURPOSE: To compare the efficacy of dorsal penile nerve block (DPNB) and eutectic mixture of local anesthetic (EMLA) for attenuation of neonatal pain during circumcision. METHOD: A total of 20 infants born at a United States upper Midwestern hospital were involved in the study. Measurements of blood pressure, heart rate, respiratory rate, and oxygen saturation were obtained along with a Neonatal Infant Pain Scale (NIPS) grading at five separate intervals (baseline, restraint, incision, Gomco clamp application, and post circumcision) throughout the circumcision procedure. A comparison was done between the two groups regarding response to the noxious stimuli. FINDINGS: Infants demonstrate physiological and behavioral response to pain. These physiological and behavioral responses are observable and measurable. In addition, results show less response with the DPNB as compared to the EMLA. CONCLUSIONS: There is a trend toward better pain control with the DPNB as compared to EMLA. PMID- 9849274 TI - ARDS: are we winning at last? PMID- 9849275 TI - Classification trees. A possible method for iso-resource grouping in intensive care. AB - Classification and grouping of clinical data into defined categories or hierarchies is difficult in intensive care practice. Diagnosis-related groups are used to categorise patients on the basis of diagnosis. However, this approach may not be applicable to intensive care where there is wide heterogeneity within diagnostic groups. Classification tree analysis uses selected independent variables to group patients according to a dependent variable in a way that reduces variation. In this study, the influence of three easily identified patient attributes on their length of intensive care unit stay was explored using classification analysis. Two thousand five hundred and forty-five critically ill patients from three hospitals were classified into groups so that the variation in length of stay within each group was minimised. In 23 out of 39 terminal groups, the interquartile range of the length of stay was < or = 3 days. PMID- 9849276 TI - The impact of a high-dependency unit on the workload of an intensive care unit. AB - While there has been recent support for high-dependency unit development, there are few data reporting the impact of such development on existing critical care facilities. Therefore the aim of this study was to examine the workload and capacity constraints of an adult general intensive care unit before and after the development of an adjacent high-dependency unit. Following the opening of the high-dependency unit, the total number of patients admitted increased by 49%. On the high-dependency unit, more elderly patients were admitted for longer and more frequently following midweek elective surgery. On the intensive care unit, patients' initial severity of illness was lower and their duration of admission decreased; fewer patients were admitted directly from the general wards. The financial benefits of high dependency care may be eroded by the increased use of the critical care services. PMID- 9849277 TI - Induced hypothermia in the postoperative management of refractory cardiac failure following paediatric cardiac surgery. AB - Postoperative low cardiac output states are a major cause of postoperative mortality in infants and children following corrective cardiac surgery for congenital heart defects. In this unit, whole body hypothermia has been used since 1979 in the management of these low output states when they are refractory to conventional modes of therapy. Twenty cases treated in this way between July 1986 and June 1990 were reviewed in 1992. The current report reviews the 50 further cases treated with moderate hypothermia between July 1990 and December 1995. The median (range) age of patients was 8 months (0 days-16 years) with a median weight of 4.1 kg (2.5-33 kg). Following cooling, there was a decrease in heart rate (p < 0.001), an increase in mean arterial pressure (p < 0.001) and a decrease in mean atrial pressure (p < 0.001). Significant increases in pH and urine output were also noticed, the increase in urine output being greater in the surviving group (p = 0.02). A decrease in platelet count occurred (p < 0.001) but white blood cell count remained unchanged (p = 0.18). Twenty-five of the 50 patients survived to leave hospital. Induced hypothermia does not appear to be associated with any complications and after the failure of all conventional treatment, it seems likely that the technique may have been beneficial to outcome in some patients. PMID- 9849278 TI - Lung aeration and pulmonary gas exchange during lumbar epidural anaesthesia and in the lithotomy position in elderly patients. AB - We investigated a total of 36 subjects with a mean (SD) age of 65 (13) years, during baseline conditions (supine, before any anaesthesia), and then during one of the following protocols: (1) lithotomy positioning (n = 12), (2) epidural anaesthesia (n = 12), (3) general anaesthesia in the supine position (n = 12). Lung aeration, ventilation/perfusion matching, gas exchange and functional residual capacity were measured. Lung aeration was normal during baseline assessment with almost no regions with poor aeration and no substantial dependent densities. Shunt and perfusion of poorly ventilated regions were minor. Lithotomy positioning did not reduce functional residual capacity and did not affect aeration of the lung or ventilation/perfusion matching. Epidural anaesthesia, in general, had no effect on aeration, ventilation/perfusion matching or gas exchange, regardless of whether the patient was in the supine or lithotomy position. General anaesthesia, however, caused significant increases in poorly aerated lung regions and in dependent densities (interpreted as atelectasis). In conclusion, no or little impairment of lung aeration and ventilation/perfusion matching was caused by the lithotomy position and/or epidural anaesthesia, contrary to the effects seen during general anaesthesia. However, our findings also suggest that being overweight is a factor that may cause impairment of lung aeration. PMID- 9849279 TI - The uptake of sevoflurane during anaesthesia. AB - The rate of uptake of sevoflurane during clinical anaesthesia (1.3 MAC) was measured by computer-controlled injection of liquid anaesthetic into a closed breathing system. The cumulative uptake of sevoflurane was 4.8 ml, 7.4 ml, 9.5 ml and 11.5 ml at 30, 60, 90 and 120 min, respectively. The ratio of inspired to end expired sevoflurane was greater than similar measurements we have made for desflurane in the past, but the absolute rate of sevoflurane uptake was less than the rate of uptake of desflurane in these cases. The rate of uptake was equivalent to 059e-0.32t + 0.039e-0.036t + 0.105e-0.0034t ml.min-1 liquid sevoflurane. Plasma urea and creatinine measured on the first postoperative day were not significantly different from pre-operative values. PMID- 9849280 TI - Comparison of rocuronium and suxamethonium for use during rapid sequence induction of anaesthesia. AB - This study was designed to compare the tracheal intubating conditions during a rapid sequence induction of anaesthesia using rocuronium 0.6 (n = 61) or 1.0 mg.kg-1 (n = 130) or suxamethonium 1.0 mg.kg-1 (n = 127) as the neuromuscular blocking drugs. Anaesthesia was induced with fentanyl 1-2 micrograms.kg-1 and thiopentone 5 mg.kg-1 (median dose) and intubating conditions were assessed 60s after the administration of the neuromuscular blocking drug by an observer unaware of which drug had been given. Intubating conditions were graded on a three-point scale as excellent, good or poor, the first two being considered clinically acceptable. The study was carried out in two parts. At the end of the first part a comparison between the two doses of rocuronium was carried out when at least 50 patients had been enrolled in each group. The results showed the intubating conditions to be significantly superior with the 1.0 mg.kg-1 dose of rocuronium (p < 0.01). Final comparison between the 1.0 mg.kg-1 doses of rocuronium and suxamethonium showed no significant difference in the incidence of acceptable intubations (96 and 97%, respectively). The incidence of excellent grade of intubations was, however, significantly higher with suxamethonium (80% vs. 65%; p = 0.02). It is concluded that rocuronium 1.0 mg.kg-1 can be used as an alternative to suxamethonium 1.0 mg.kg-1 as part of a rapid sequence induction provided there is no anticipated difficulty in intubation. The clinical duration of this dose of rocuronium is, however, 50-60 min. PMID- 9849282 TI - The effect of intrathecal administration of magnesium sulphate in rats. AB - Somatosensory evoked potential, locomotion and vocalisation upon tail pinch in rats was studied in order to determine whether intrathecal magnesium sulphate administration causes spinal anaesthesia. In Wistar rats with indwelling intrathecal catheters, cortical somatosensory evoked potential was recorded following stimulation via electrodes inserted into the hind paw under chloral hydrate anaesthesia before and after intrathecal administration of 10 microliters of either magnesium sulphate (12.3% or 24.6%) or lignocaine (4% or 8%). Locomotion and vocalisation after tail pinch were tested following intrathecal administration of the same two drugs in conscious rats. Somatosensory evoked potential amplitude was diminished after administration of lignocaine (p < 0.05) but did not change after magnesium sulphate. Latency of P1 was increased by lignocaine and by magnesium sulphate 12.3% (p < 0.05). Although lower extremity paralysis was observed in both groups, its duration with magnesium sulphate was much longer than with lignocaine. Vocalisation was recognised after magnesium sulphate 12.3%, but was not observed after lignocaine 8% during paralysis (p < 0.05). We believe that magnesium sulphate caused motor paralysis, but not complete analgesia. PMID- 9849281 TI - Neuromuscular interaction between cisatracurium and mivacurium, atracurium, vecuronium or rocuronium administered in combination. AB - We compared the dose-response relationships of cisatracurium, mivacurium, atracurium, vecuronium and rocuronium and examined the interactions of cisatracurium with mivacurium, atracurium, vecuronium and rocuronium in humans by isobolographic and fractional analyses. We studied 180 adult patients during nitrous oxide-fentanyl-propofol anaesthesia. Neuromuscular block was monitored using mechanomyography to detect the twitch response of the ulnar nerve at the wrist. The dose-response curves were determined by probit analysis. The calculated ED50 values and their 95% confidence intervals were 40.9 (38.1-43.7), 49.8 (47.0-52.6), 187.2 (175.1-199.3), 36.6 (34.7-38.5) and 136.4 (129.2-143.6) micrograms.kg-1 for cisatracurium, mivacurium, atracurium, vecuronium and rocuronium, respectively. Corresponding ED95 values were 57.6 (53.5-61.7), 91.8 (88.1-95.5), 253.1 (238.9-267.3), 52.9 (49.1-56.7) and 288.7 (276.2-301.2) micrograms.kg-1, respectively. The interaction between cisatracurium and mivacurium, vecuronium or rocuronium was found to be synergistic, but the interaction between cisatracurium and atracurium was found to be additive. Synergy between cisatracurium and vecuronium or rocuronium was greater than between cisatracurium and mivacurium. PMID- 9849283 TI - Revised checklist for anaesthetic machines. AB - A revised edition of the guidelines of the Association of Anaesthetists of Great Britain and Ireland, for the pre-operative check of anaesthetic machines, was published in March 1997. A checklist based on the revised guidelines was used for the routine pre-operative checks of anaesthetic machines over a 6-week period in a district general hospital. One hundred and thirty-two checklists were completed. These were analysed for the time taken to complete the check and for the faults found in the anaesthetic machines. The mean time taken to complete a check was 6.8 min and the mean time taken to complete two consecutive checks, in the anaesthetic room and operating theatre, was 12.7 min. Carbon dioxide cylinders were present on the machines in 99 checks (75%), contrary to Association guidelines. Other faults were found in 40 checks (30.3%). The most frequent cause of faults was the oxygen analyser, faults being found in 15 checks. Other frequent faults were due to empty vaporisers or spare gas cylinders and the emergency oxygen bypass control. PMID- 9849284 TI - Fibreoptic intubation using the cuffed oropharyngeal airway and Aintree intubation catheter. AB - A cuffed oropharyngeal airway has recently been introduced which has larger internal dimensions than a comparable Guedel airway. This allows a ventilation/exchange bougie, the Aintree Intubation Catheter, mounted on a fibreoptic laryngoscope to pass through it. Its 15-mm connector and pharyngeal cuff suggested the possibility of using a Rusch sealed-port angle piece to allow ventilation through the oropharyngeal airway during fibreoptic laryngoscopy. This study investigated using this equipment to intubate the trachea through the cuffed oropharyngeal airway in paralysed patients, whilst maintaining ventilation manually with a Bain system. In 20 patients, airway control was satisfactory throughout and tracheal intubation was accomplished without complications. The cuffed oropharyngeal airway was easy to manipulate to improve a suboptimal fibreoptic view of the larynx. This may give it an advantage over the laryngeal mask airway when used as a ventilation/intubation conduit. PMID- 9849285 TI - Hypophosphataemia. Pathophysiology, effects and management on the intensive care unit. AB - Routine detection and treatment of hypophosphataemia on the intensive care unit is commonplace. Hypophosphataemia has been associated with a multitude of clinical effects and there are many associations between correction of hypophosphataemia and improvement in symptoms. However, there is no evidence at present to support the routine correction of hypophosphataemia in the absence of clinical symptoms or signs. PMID- 9849286 TI - Use of the McCoy laryngoscope or fingers to facilitate fibrescope-aided tracheal intubation. AB - In fibrescope-aided tracheal intubation, it can be difficult to advance a tube over a fibrescope, because its passage may be impeded by the epiglottis, arytenoids or pyriform fossa. In two patients with difficult intubation, after successful insertion of a fibrescope into the trachea, it was impossible to advance the tube over the fibrescope. Fibrescope-aided tracheal intubation was accomplished by shifting the epiglottis anteriorly and 'opening up' the glottis either by fingers or the McCoy laryngoscope. PMID- 9849287 TI - Gallstones and isoflurane hepatitis. AB - This report is of a case of a previously fit 65-year-old woman who developed postoperative liver dysfunction following an anaesthetic involving isoflurane. Biliary ultrasound demonstrated gallstones. However, serum antibodies to trifluoroacetylated proteins were detected, suggesting that immune sensitisation to the anaesthetic could have contributed to the impaired liver function. PMID- 9849288 TI - Postoperative nausea and vomiting in arthroscopic day-case surgery: a comparison between desflurane and isoflurane. AB - We compared the postoperative effects, in particular nausea and vomiting, of two commonly used anaesthetic agents, isoflurane and desflurane, in patients undergoing day-case arthroscopic surgery. We found a significantly higher incidence of postoperative nausea and vomiting in those patients receiving desflurane. In our study, desflurane was associated with a higher incidence of peroperative respiratory complications, including airway irritability and coughing. PMID- 9849289 TI - Anaesthesia for video-assisted thoracoscopic patent ductus arteriosus ligation. AB - The case notes of children undergoing video-assisted thoracoscopic ligation of a patent ductus arteriosus were reviewed with particular emphasis on the anaesthetic management. All children were managed using one-lung ventilation with no serious adverse sequelae. The lungs could be isolated easily in all cases and no special equipment was required. We describe our initial series of 13 cases and discuss the anaesthetic implications arising from this surgical technique. We also discuss the different ways of monitoring duct closure. PMID- 9849290 TI - The effect of acupuncture on the incidence of postextubation laryngospasm in children. AB - Laryngospasm occurring after tracheal extubation in children is potentially dangerous. This study uses acupuncture with bloodletting at the Shao Shang (L 11) or Shang Yang (Li 1) acupoints to investigate whether this technique can prevent or treat laryngospasm. Seventy-six patients were randomly divided into two groups. Patients in the acupuncture group (n = 38) were treated with bilateral Shao Shang acupunctures at the end of the operation. Patients in the control group (n = 38) were not. The incidence of laryngospasm in the acupuncture group (5.3%) was less than that in the control group (23.7%) (p < 0.05). If laryngospasm developed, patients were immediately treated with acupuncture at either the Shao Shang or Shang Yang acupoints. As judged by an increase in peripheral oxygen saturation, the laryngospasm was relieved within 1 min of acupuncture in all patients. It is concluded that acupuncture with bloodletting at the Shao Shang acupoint may prevent and treat laryngospasm occurring after tracheal extubation in children. PMID- 9849291 TI - Gastro-oesophageal reflux during day case gynaecological laparoscopy under positive pressure ventilation: laryngeal mask vs. tracheal intubation. AB - This study aimed to evaluate whether or not the use of intermittent positive pressure ventilation via the laryngeal mask airway is associated with a higher risk of gastro-oesophageal reflux when compared with intermittent positive pressure ventilation via a tracheal tube in patients undergoing day case gynaecological laparoscopy in the head down position. Sixty healthy women were randomly allocated to receive either the laryngeal mask or cuffed tracheal tube for intra-operative airway maintenance. Using continuous oesophageal pH monitoring, four patients in the tracheal tube group and none in the laryngeal mask group had evidence of gastro-oesophageal reflux (as indicated by a decrease in oesophageal pH to below 4). The difference in the incidence of reflux did not achieve statistical significance (p = 0.11). In conclusion, we found no evidence to suggest that the use of intermittent positive pressure ventilation via the laryngeal mask increases the risk of gastro-oesophageal reflux in patients undergoing elective day case gynaecological laparoscopy. PMID- 9849292 TI - Dorsal column function after spinal/epidural anaesthesia. PMID- 9849293 TI - Are you all right, Mrs Tilt? PMID- 9849294 TI - Caudal epidurals: the 'whoosh test'. PMID- 9849295 TI - Iatrogenic puncture of the laryngeal mask airway cuff. PMID- 9849296 TI - The intubating laryngeal mask airway (ILMA) in failed and difficult intubation. PMID- 9849297 TI - Ethics and research in anaesthesia. PMID- 9849298 TI - Acute arrhythmias on induction of anaesthesia in a child with a blocked shunt. PMID- 9849299 TI - Ventilation during percutaneous tracheostomy. PMID- 9849300 TI - Percutaneous dilatational tracheostomy in children. PMID- 9849301 TI - Dangers of using a pneumatic pressure bag to increase transfusion rate. PMID- 9849302 TI - Pain and venous cannulation. PMID- 9849303 TI - Accuracy of bedside haemoglobin measurement. PMID- 9849304 TI - The influence of smoking on postoperative nausea and vomiting. PMID- 9849305 TI - The Budget: bad news for British anaesthetists in the USA. PMID- 9849306 TI - Hyperlipidaemia in hyperuricaemia and gout. PMID- 9849307 TI - Early treatment of rheumatoid arthritis: rationale, evidence, and implications. PMID- 9849308 TI - Lymphadenopathy in a patient with systemic onset juvenile chronic arthritis. PMID- 9849309 TI - Osteoarthritis of the hip: agreement between joint space width measurements on standing and supine conventional radiographs. AB - OBJECTIVE: To assess the effect of standing position on joint space width (JSW) measurements of the hips with and without osteoarthritis (OA) on pelvic radiographs. METHODS: Adult patients aged 18 or more had pelvic anteroposterior conventional radiographs standing and supine performed by a single radiologist in the same radiology unit according to standardised guidelines. JSW measurements in mm were made by a single reader blind to patients' identity and type of view, using a 0.1 mm graduated magnifying glass directly laid over the radiograph, at the narrowest point for OA hips or at the vertical joint space for non-OA hips. Agreement of JSW between both views was assessed using the Bland and Altman graphical analysis. RESULTS: JSW was greater on standing than supine radiographs, for example, 7.1% for OA hips. Mean (SD) differences and limits of agreement (mm) between both views were 0.08 (0.27) and -0.46 to 0.62 for the 70 non-OA hips, 0.02 (0.31) and -0.60 to 0.64 for the 46 OA hips. Corresponding 95% confidence intervals of mean difference were 0.02, -0.14 mm and -0.07, -0.11 mm. CONCLUSIONS: Measurements of JSW of the hip on pelvic standing and supine radiographs are concordant. Changes less than or equal to 0.64 mm between the two views are similar or inferior to radiological progression of OA. PMID- 9849310 TI - Evidence for genetic anticipation in nodal osteoarthritis. AB - OBJECTIVE: Evidence was sought for genetic anticipation (disease occurring at an earlier age in subsequent generations, with increasing severity) in nodal osteoarthritis (NOA). METHODS: Age at symptom onset and disease severity was compared within 30 parent/offspring pairs with NOA. Correlation between the offspring age of disease onset and the parental age at conception was also assessed. RESULTS: The age at onset of nodal symptoms was earlier in the offspring (43 years (95% confidence intervals (CI) 38 to 47) v 61 (CI 58 to 65); mean difference 18 years (CI 13 to 22): p < 0.001) as was larger joint symptom onset (48 years (CI 41 to 55) v 67 (CI 61 to 73); mean difference 20 years (CI 13 to 27): p < 0.01). A negative correlation existed between age of offspring symptom onset and parental age at conception. Fifteen (50%) offspring had similar or more extensive disease than their parents. CONCLUSIONS: These results suggest genetic anticipation occurs in NOA and if confirmed a search for trinucleotide repeats is warranted. PMID- 9849311 TI - Serum concentrations of cartilage oligomeric matrix protein and bone sialoprotein in hip osteoarthritis: a one year prospective study. AB - OBJECTIVE: To evaluate serum concentrations of cartilage oligomeic matrix protein (COMP) and bone sialoprotein (BSP) as predictors of disease progression in hip osteoarthrtitis (OA). METHODS: Forty eight consecutive patients, referred to hospital for symptomatic hip OA, (ACR criteria) were monitored in a one year prospective trial with radiographs and serum samples. The radiographs were graded for joint space narrowing, osteophytes, and sclerosis and the joint space width was measured by a digitised image analyser. Serum COMP and BSP were quantified by immunoassays. RESULTS: The COMP concentrations at baseline correlated with the joint space width at entry and with its yearly mean narrowing (r = 0.38, p = 0.002) but not with joint space narrowing grade progression. The concentrations were higher in patients with bilateral hip OA (p = 0.03). The serum BSP concentrations at baseline were unrelated to OA progression but correlated inversely to the osteophyte grade (r = -0.36, p = 0.004) and sclerosis grade (r = -0.42, p = 0.0004). CONCLUSION: Serum COMP seems to be a surrogate marker of OA and may be of interest for the detection of patients at risk of rapidly progressing disease in hip OA. Serum BSP changes seem to reflect alterations in the subchondral bone turnover in hip OA. Measurement of joint space width using a digitised image analyser is a sensitive way of assessing OA progression that facilitates evaluation of tissue markers in relation to anatomical changes in the joint. PMID- 9849312 TI - Only high disease activity and positive rheumatoid factor indicate poor prognosis in patients with early rheumatoid arthritis treated with "sawtooth" strategy. AB - OBJECTIVES: To investigate the prognostic significance of clinical and genetic markers on the outcome of patients with recent-onset rheumatoid arthritis (RA) treated actively with slow acting antirheumatic drugs (SAARDs). METHODS: A total of 142 consecutive patients with early RA (median disease duration of 7 months) were treated according to the "sawtooth" strategy and prospectively followed up for an average of 6.2 years. Several clinical parameters at start as well as genetic markers were related to the functional outcome (ARA Functional class and HAQ disability score) and radiographic joint damage (Larsen's score) at the latest visit. RESULTS: In logistic regression analysis only Mallya score (including morning stiffness, pain scale, grip strength, Ritchie's articular index, haemoglobin, and erythrocyte sedimentation rate) at baseline, and Mallya score and rheumatoid factor (RF) positivity at one year were found to be of significance with respect to the radiographic outcome of the patients. Furthermore, at the latest visit HAQ score was related to radiographic score. At baseline the mean ages of the DR4 positive patients and the patients with RA associated DR alleles were statistically significantly lower than those without the above mentioned risk factors (44 v 49, p = 0.03 and 41 v 53, p = 0.04, respectively). However, these genetic markers had no prognostic significance on the functional or radiographic outcome of the patients. CONCLUSION: High clinical disease activity at baseline and RF positivity especially at one year after the institution of SAARD treatment are the best predictors of poor prognosis in early RA. However, from the clinical point of view, the disease outcome of an individual patient with early RA, cannot be predicted accurately enough by present means. PMID- 9849313 TI - Deforming arthropathy or lupus and rhupus hands in systemic lupus erythematosus. AB - OBJECTIVE: Although deforming arthropathy in systemic lupus erythematosus (SLE) is characterised by a number of manifestations, definitive criteria for the different forms have not yet been established. To define deforming arthropathy and its different types a study was undertaken of 176 SLE patients. METHODS: Using as criterion any deviation from any of the metacarpus finger axes 17 patients (16 women, one man) were identified with clinical deforming arthropathy. These patients were evaluated according to a standardised protocol that covered all known characteristics of deforming arthropathy. By means of "Jaccoud's arthropathy index" three different forms were identified. RESULTS: Three patients had an erosive form of deforming arthropathy (or rhupus hand) such as those seen in frank rheumatoid arthritis (RA), eight patients were identified as having Jaccoud's arthropathy (or lupus hand), and the remaining six patients had mild deforming arthropathy. Jaccoud's arthropathy is characterised by severe deformation of the hands (ulnar deviation, swan neck deformities, and Z deformity of the thumb) and feet with multiple non-erosive subluxations, mild aching and little or no evidence of synovitis. All patients, but one, fulfilled just four criteria of the ACR classification and joint symptoms were always found to precede the diagnosis of SLE. Furthermore a remarkable association of Jaccoud's arthropathy with fetal loss, thrombosis--both venous and arterial--and the presence of antiphospholipid antibodies was found. CONCLUSIONS: These data suggest that Jaccoud's arthropathy represents a subset of SLE. Subdivision of deforming arthropathy into several clinical forms can facilitate the clinical management of this disorder. PMID- 9849314 TI - Efficacy of allopurinol and benzbromarone for the control of hyperuricaemia. A pathogenic approach to the treatment of primary chronic gout. AB - OBJECTIVES: To study the efficacy of allopurinol and benzbromarone to reduce serum urate concentrations in patients with primary chronic gout. METHODS: Prospective, parallel, open study of 86 consecutive male patients with primary chronic gout. Forty nine patients (26 normal excretors and 23 under excretors) were given allopurinol 300 mg/day and 37 under excretors benzbromarone 100 mg/day. After achieving steady plasma urate concentrations with such doses, treatment was then adjusted to obtain optimal plasmatic urate concentrations (under 6 mg/dl). RESULTS: Patients receiving allopurinol 300 mg/day showed a mean reduction of plasmatic urate of 2.75 mg/dl (from 8.60 to 5.85 mg/dl) and 3.34 mg/dl (from 9.10 to 5.76 mg/dl) in normal excretors and under excretors respectively. Patients receiving benzbromarone 100 mg/day achieved a reduction of plasmatic urate of 5.04 mg/dl (from 8.58 to 3.54 mg/dl). Fifty three per cent of patients receiving allopurinol and 100% receiving benzbromarone achieved optimal plasma urate concentrations at such doses. The patients with poor results with allopurinol 300 mg/day achieved a proper plasma urate concentration with allopurinol 450 to 600 mg/day, the mean final dose being 372 mg/day. Renal function improved and no case of renal lithiasis was observed among benzbromarone treated patients, whose mean final dose was 76 mg/day. CONCLUSION: Benzbromarone is very effective to control plasma urate concentrations at doses ranging from 50 to 100 mg/day. Uricosuric treatment is a suitable approach to the treatment of patients with gout who show underexcretion of urate. PMID- 9849317 TI - Correlation of 9G4 idiotope with disease activity in patients with systemic lupus erythematosus. AB - OBJECTIVE: To compare the levels of the 9G4 idiotope (9G4 Id) in systemic lupus erythematosus (SLE) patients with a detailed disease activity index, the British Isles Lupus Assessment Group (BILAG) index, and serological parameters of disease activity by ds DNA antibody levels and serum C3 concentrations. METHODS: In a cross sectional analysis serum samples from 190 patients with SLE were studied and a further 55 serial bleeds from 14 patients. An enzyme linked immunosorbent assay was used to measure the 9G4 Id, and anti dsDNA and antimyeloperoxidase (MPO) antibodies. The C3 levels were measured by laser nephelometer. RESULTS: Seventy six of 190 (40%) of the patients tested had raised 9G4 Id levels. In the cross sectional study 9G4 Id levels were found to correlate with disease activity in the BILAG cardiovascular/respiratory renal, and haematological systems and with global BILAG score (p < 0.01). In the serial bleeds 9G4 Id levels correlated with anti-dsDNA antibody and C3 levels, but not with anti-MPO antibodies. No correlations were found with treatment. In six cases the 9G4 Id levels correlated well with global BILAG scores and dsDNA antibody levels. In four cases the BILAG global and 9G4 Id levels alone correlated well. CONCLUSIONS: Raised levels of the 9G4 Id are present in a substantial proportion of serum samples from patients with lupus, correlate with various aspects of disease activity in SLE. The Id is detectable on anti-dsDNA antibodies, though it must also be present on other immunoglobulins whose specificities remain unknown. PMID- 9849315 TI - Increased expression of human type IIa secretory phospholipase A2 antigen in arthritic synovium. AB - OBJECTIVE: To determine the localisation and level of expression of human type IIa secretory phospholipase A2 (sPLA2) in the synovium of rheumatoid arthritis (RA), osteoarthritis (OA), and non-arthritic (NA) patients and to examine the relation between sPLA2 and histological features of inflammation. METHODS: Immunoperoxidase staining using the anti-sPLA2 monoclonal antibody 9C1 was performed on frozen sections of knee synovium of 10 RA, 10 OA, and 10 NA patients. sPLA2 positive cells were scored on a scale of 0-3 in 10 fields of a representative tissue section from each case. Double labelling immunofluorescence confocal microscopy with antibodies to CD14 or CD45 and 9C1 was used to determine cell type specificity. Inflammation was assessed by semiquantitative scoring of lining layer thickness and mononuclear cell infiltrates (MC) and a cumulative inflammation score, generated by summing the two parameters. Scores in each group were compared using non-parametric statistical analysis. RESULTS: sPLA2 was localised to endothelium (EC), vascular smooth muscle (VSM), and mast cells (M) in all tissue sections. In RA and OA sections, staining was seen in both macrophage-like and fibroblast-like cells in the synovial lining layer (LL) and subsynovial lining layer (SLL). Perineural cells stained positively. Subintimal lymphoid aggregates (LA) were negative in all sections. The RA group showed significantly greater staining in extravascular synovial tissue (median 3.6, range 1.5-6.0) than the OA (median 1.95, range 0-5.3) or NA (median 0, range 0 5.9) groups (p < 0.05). LL staining was significantly higher in RA than both OA and NA sections (p < 0.05). The OA group showed a trend to higher staining scores than the NA group that did not reach significance. There was a significant correlation between the sPLA2 staining score and inflammation score within the RA patient group (p < 0.05). CONCLUSIONS: The synovium is a site of increased expression of sPLA2 antigen in both RA and OA relative to NA. Its presence in both fibroblast and macrophage-like cells in the LL and SLL of synovial tissue in RA and OA, but not NA, indicates that the enzyme is specifically induced in these regions in both conditions with expression in the LL being particularly characteristic of RA. The widespread expression of sPLA2 in synovium suggests it is likely to play a significant part in synovial pathology. PMID- 9849318 TI - Renal involvement in Chinese patients with rheumatoid arthritis. PMID- 9849316 TI - Loss of laminin and of the laminin receptor integrin subunit alpha 6 in situ correlates with cytokine induced down regulation of alpha 6 on fibroblast-like synoviocytes from rheumatoid arthritis. AB - OBJECTIVE: To investigate in situ the expression of the integrin receptor subunits alpha 6 and beta 1 and the distribution of the ligand laminin in the synovia from osteoarthritis (OA) and rheumatoid arthritis (RA) patients and to study the effect of cytokines and antirheumatic drugs on the expression of the alpha 6 and beta 1 integrin subunits on long term cultures of fibroblast-like synoviocytes (FBS) derived from OA and RA. METHODS: The expression of the alpha 6 and beta 1 integrin subunits and the distribution of laminin were examined immunohistochemically in normal synovia and in synovia from patients with OA and RA. The effect of proinflammatory cytokines (IL1 beta and TNF alpha), and of antirheumatic drugs (salicylic acid, dexamethasone, and methotrexate) on the alpha 6 and beta 1 expression of cultured normal FBS and FBS from patients with OA and RA was determined by flow cytometry. RESULTS: In normal synovia and in OA synovia samples with a low grade of inflammation, synovial lining cells (SLC) showed a parallel expression and distribution of alpha 6 and laminin. In synovia samples of OA with a higher grade of inflammation and in the majority of RA synovia samples laminin was pericellularly distributed in a low number of SLC, whereas alpha 6 was expressed on the surface of a high number of SLC. In RA synovia samples with severe inflammatory changes the gradual loss of laminin generally corresponded to a decrease of the alpha 6 integrin subunit. beta 1 was always strongly expressed in all synovia samples detected. Proinflammatory cytokines up regulated the expression of alpha 6 and beta 1 on OA-FBS, whereas these effectors decreases alpha 6 and beta 1 on RA-FBS. In contrast, antirheumatic drugs, in particular methotrexate and dexamethasone, reduced the expression of alpha 6 and beta 1 on OA-FBS, whereas the same treatment on RA-FBS stimulated the expression of these integrin subunits. CONCLUSION: The gradual loss of laminin in chronic synovitis may contribute to the altered expression of alpha 6 in SLC. IL1 beta and TNF alpha down regulated the expression of the alpha 6 and beta 1 integrin subunits on long term cultures of FBS derived from RA. Therefore, these cytokines may be among the effectors regulating the expression of the alpha 6 integrin subunit in SLC in vivo. As antirheumatic drugs increase the expression of alpha 6 on RA-FBS, the presence of the laminin receptor may confer a protective effect on the synovia in vivo. PMID- 9849319 TI - Evaluation of adverse experiences related to pamidronate infusion in Paget's disease of bone. PMID- 9849321 TI - Are basic surgical trainees being 'short-changed'? PMID- 9849320 TI - The MRCS written examination: a new philosophy. PMID- 9849322 TI - Medical negligence and justice: the Lord Chancellor explains the latest developments. PMID- 9849323 TI - Pre-operative advice given by general practitioners and consultants to patients concerning return to work. PMID- 9849324 TI - Orthopaedic pre-operative assessment: a two-year experience in 5,000 patients. PMID- 9849325 TI - The reduction in hours of work for surgical trainees--an enlightened move or a great mistake? PMID- 9849326 TI - Who audits the auditors? PMID- 9849327 TI - The role of the nurse in the pre-admission clinic. PMID- 9849328 TI - Primary hyperparathyroidism: a surgical perspective. PMID- 9849329 TI - Pancreas transplantation. PMID- 9849330 TI - Haemorrhagic complications of pancreatitis: presentation, diagnosis and management. AB - Massive haemorrhage is an uncommon complication in pancreatitis. Most affected patients suffer from chronic disease with associated pseudocyst. We present five patients (four male) with a mean age of 41 years (range 34-48 years). All patients had alcohol-induced pancreatitis complicated either by haematemesis (3), intraperitoneal haemorrhage (1) or both haematemesis and intraperitoneal haemorrhage (1). Source of bleeding was pseudocyst wall (2), splenic artery pseudoaneurysm (2) and splenic artery rupture (1). Distal pancreatectomy and splenectomy was performed in two patients, intracystic ligation and drainage in two, and packing with subsequent external drainage in one. Rebleeding occurred in two patients and required subsequent distal pancreatectomy and splenectomy in one; the other patient died of splenic rupture. No rebleeding and no mortality occurred after resection. Primary pancreatic resection is recommended whenever possible. Other management options include embolisation and ligation. PMID- 9849331 TI - Periampullary diverticula: consequences of failed ERCP. AB - Periampullary diverticula (PAD) are associated with biliary disease and contribute to failure of endoscopic retrograde cholangiopancreatography (ERCP), especially in elderly patients. The presence of PAD and causes of failure to cannulate the ampulla were noted in 1211 consecutive patients undergoing ERCP. Case notes of 100 consecutive patients with PAD were reviewed retrospectively. Overall prevalence of PAD was 9%. Prevalence was higher in patients > or = 75 years when compared with those < 75 years (19.2% vs 4.8%, P < 0.0001). Ampullary cannulation was successful in 62.4% of patients with PAD and 92.7% without PAD (P < 0.0001). Success rates were lower in patients with intradiverticular papillae than in those with juxtapapillary diverticula (38.1% vs 77.6%; P < 0.0001). Of 19 patients with PAD who did not have any imaging other than ultrasound, 16 were asymptomatic over a median follow-up of 20 months. Biliary surgery was performed on 35 patients, with no major complication. PAD are a major cause of failed ERCP. Failure rates are higher in patients with intradiverticular papillae than juxtapapillary diverticula. Though a large proportion of patients not imaged remain asymptomatic on follow-up, it is difficult to predict which patients may form this group. Surgery, when indicated, is safe and effective in elderly patients in whom ERCP has failed. PMID- 9849332 TI - Primary small intestinal tumours: increased incidence of lymphoma and improved survival. AB - Small intestinal malignancies are rare and may have a delayed presentation owing to insidious growth. We have reviewed the case notes of 25 patients presenting with primary small bowel tumours over a 10-year period. Abdominal pain, weight loss and vomiting were the most common symptoms. The median duration of symptoms was 6 months. Physical examination was normal in 24% of patients. An abdominal mass was present in 46% of cases. Emergency laparotomy was undertaken in 28% of patients. Lymphomas were identified in 72% and adenocarcinomas were present in 16%. The predominance of small bowel lymphoma is an unusual finding and may be related to the high incidence of coeliac disease in the region. The median survival in the lymphoma group was 36 months, which compares favourably with reported series. PMID- 9849333 TI - Adult hernia surgery in Wales revisited: impact of the guidelines of The Royal College of Surgeons of England. AB - This study investigated the impact of the guidelines of The Royal College of Surgeons of England on the practice of hernia surgery in Wales. This was assessed by means of a postal survey to all consultant general surgeons in Wales in 1996 1997. The areas covered were: awareness of the guidelines of The Royal College of Surgeons of England and the impact of such guidelines on their practice, attendance at hernia courses, operative technique, materials used for repair and skin suture, proportion of day case hernias, length of inpatient stay, thromboembolic (TE) prophylaxis and postoperative advice to patients with regard to light work, heavy work and sport. In all, 79 replies were received (85%). Almost all the surgeons had read the guidelines; this changed the practice of 20% of respondents but did not in 32%. A further 48% did not answer the question. In contrast with our 1993 survey results, in Wales there is now a uniform surgical management of adult inguinal hernias: the most common operation is the Liechtenstein, with monofilament non-absorbable suture to secure the mesh, followed by the Shouldice repair. The Bassini and inguinal darn operations are becoming much less common and none now uses braided or absorbable sutures for the repair. Skin closure is still rather variable, with only 58% of respondents adhering to the recommended absorbable subcuticular suture. Postoperative advice is now uniform and in accordance with the guidelines. A trend towards more TE prophylaxis and more day case hernia surgery is also seen. PMID- 9849334 TI - Mortality after elective abdominal aortic aneurysm repair: not where ... but how many and by whom. PMID- 9849335 TI - Screening for abdominal aortic aneurysms does not increase psychological morbidity. AB - Screening can lead to harmful psychological effects in the screened population- an argument used against abdominal aortic aneurysm (AAA) screening. However, there is no evidence for this in AAA screening. We applied the Hospital Anxiety and Depression Scale (HADS) to a group of men undergoing screening for AAA. The HADS questionnaire was completed by subjects found not to have AAA, subjects with known small aneurysms attending for follow-up scans, subjects with known AAA on waiting lists for surgery, and controls not involved in the screening programme. The groups were well matched for age and the number of additional diseases. There was no significant difference in the distribution of patients for anxiety and depression according to the HADS questionnaire (chi 2 test, P > 0.1). The results from this study suggest that AAA screening does not increase anxiety or depression in the screened subjects--contrary to the argument put forward against screening for this condition. PMID- 9849336 TI - Systemic thrombolysis causing anastomotic dehiscence of an aortobifemoral graft. PMID- 9849337 TI - Use of a thromboembolic risk score to improve thromboprophylaxis in surgical patients. AB - Two hundred admissions to a general surgical ward were audited prospectively before and after the introduction of a thromboembolic risk score. This was based on the Thromboembolic Risk Factors (THRIFT) Consensus Group guidelines for thrombo-prophylaxis. The results showed an overall improvement in compliance from 65% to 79%. High risk patients formed 24% of the patients studied. In this group, compliance improved significantly from 14% to 58%. The overall prevalence of important thromboembolic risk factors was calculated. Of the patients, 26.5% had a Body Mass Index (BMI) of > 25, and 10% gave a past or family history of thromboembolism. Of female patients, 24% were taking oestrogens. We conclude that quantitative assessment of all patients for thromboembolic risk can improve the implementation of thromboprophylaxis, particularly in high risk patients. PMID- 9849338 TI - Current practice in primary total hip replacement: results from the National Hip Replacement Outcome Project. AB - As part of the National Study of Primary Hip Replacement Outcome, 402 consultant orthopaedic surgeons from three regions were contacted by postal questionnaire which covered all aspects of total hip replacement (THR). There was a 70% response rate of which 71 did not perform hip surgery, a further 33 refused to take part, leaving 181 valid responses. Preoperative assessment clinics were used by 89% of surgeons, but anaesthetists and rehabilitation services were rarely involved at this stage. Of respondents, 99% used routine thromboprophylaxis, with 79% using a combination of mechanical and chemical methods. Of surgeons, 84% routinely used stockings, whereas 95.5% used chemical prophylaxis, 63% employed low molecular weight heparins. Theatre facilities were shared with other surgical specialties by 6% of surgeons and 18% regularly used body exhaust suits for THR. Antibiotic loaded cement was used by 69% of surgeons, the majority (65%) used a single brand of normal viscosity cement with 9% using reduced viscosity formulations. Modern cementing techniques were commonly used at least in part, 87% used a cement gun and 94% a cement restrictor for femoral cementing. On the acetabulum, 47% pressurised the cement. In all, 36 different femoral stems and 35 acetabular cups were in routine use, but the majority of surgeons (55%) used Charnley type prostheses. Of the surgeons, 57% performed only cemented THR, while 3% exclusively used uncemented THR. Of consultants, 21% followed up their patients to 5 years, the majority discharge patients within the first year. Of concern is a large proportion of surgeons using low molecular weight heparins despite a lack of evidence with regard to reducing fatal pulmonary embolism, and also the small number of surgeons using prostheses of unproven value. Third generation cementing techniques have yet to be fully adopted. The introduction of a national hip register could help to resolve some of these issues. PMID- 9849339 TI - Management of sexually transmitted disease by surgeons. AB - The management of 63 patients diagnosed by surgeons as having sexually transmitted disease (STD) was audited. A diagnosis of STD was made in 51 (81%) of patients without taking a sexual history. Only 2 (3%) patients were referred to genitourinary medicine (GUM). Appropriate microbiological specimens were obtained from only two of 52 (4%) patients diagnosed with either pelvic inflammatory disease (PID) or epididymo-orchitis. Reliance was placed on inappropriate specimens in 22 (42%). There was widespread use of inappropriate antibiotics. The management of sexually transmitted disease by surgeons was very poor. These patients should all be referred to genito-urinary medicine. PMID- 9849340 TI - Simple aid in removal of a diastasis screw. PMID- 9849341 TI - Endoscopic sinus surgery: are junior doctors being properly trained? AB - Endoscopic sinus surgery (ESS) is a technique which carries great potential benefits for the treatment of many nasal conditions. However, it also carries substantial risks. The key to safe surgery lies with adequate training. A survey carried out as part of a North (East) Thames Region audit of higher surgical trainees revealed large discrepancies in their training and in their subsequent clinical practice. Almost half the trainees had started ESS without having been on a training course or performed any cadaver dissections. Despite the potential hazards of ESS, audit of complications and outcome received a low priority. The trainees made several suggestions for improving training, including better provision of courses, regional training programmes and improved access to cadavers for dissection. Other surgical specialties are being forced to examine the prospect of specific accreditation for minimally invasive techniques and otorhinolaryngology may have to follow suit. PMID- 9849342 TI - Twenty-four hour access to a CD-ROM surgical database has educational and patient management benefits. AB - This study determined whether 24 h access to current surgical literature via a personal computer CD-ROM was of relevance to the junior surgeon managing acute surgical referrals. Over a 2 month period, a total of 53 acute surgical referrals were assessed and initially managed by a single basic surgical trainee. The Medline database was searched using Silver Platter software for current surgical literature regarding aetiology and management after the initial patient assessment. Of the 53 searches made (mean search time 7 min 22 s), 20 (38%) were judged to have produced information that increased basic surgical trainee knowledge and a further 9 (17%) changed patient management. By contrast, 24 (45%) of searches did not reveal helpful information. The availability of a surgical database that provides the trainee with abstracts of the current literature is valuable both for the educational development of the surgeon and improvement of patient care. PMID- 9849343 TI - HRGs for benchmarking and contractual arrangements: a poisoned chalice? AB - A study was undertaken to determine if healthcare resource groups (HRGs) are reliable, reproducible in different centres and accurately reflect clinical activity. During a 3 month period, 385 finished consultant episodes (FCEs) were assessed. Of these, 17 (4.4%) were grouped to U01, i.e. no group allocated as the primary diagnosis was not recorded. For 271 of these episodes, a clinician had also allocated a choice of what he thought the HRG should be. These choices of HRG were then compared with the official grouping determined by the software provided by the National Casemix Office (NCMO). Disparities between 'automatic grouping' and proposed HRG assignment by the consultants occurred in 20.66% (56/271). Version 3 should see some of these disparities and actual grouping software errors reduced, in particular through the introduction of a new vascular surgery chapter. However, the absence of a primary diagnosis will still produce the allocation of group U01 in version 3. Caution should be observed in using the HRG software and in the interpretation of data obtained from it. This could be especially important in benchmarking or contractual settings as there may be adverse implications to the unit/hospitals involved. PMID- 9849344 TI - Simple technique for non-operative removal of ureteric stents after renal transplantation. AB - Ureteric stents are employed peroperatively in renal transplantation when there is actual or suspected damage to the donor ureter. We describe a simple technique of suturing the distal end of the stent to the tip of a urinary catheter at the time of construction of the ureteric anastomosis. The stent is simply removed by gentle traction on the urinary catheter. This method obviates the need for a minor operative procedure or exposure to ionising radiation, which are the techniques commonly utilised at present. The procedure is also applicable to reconstructive urological procedures in which short-term stenting may be required. PMID- 9849345 TI - Is cholecystectomy effective treatment for symptomatic gallstones? Clinical outcome after long-term follow-up. PMID- 9849346 TI - Laparoscopic cholecystectomy, bile duct injury and the British and Irish surgeon. PMID- 9849347 TI - Laparoscopic cholecystectomy, bile duct injury and the British and Irish surgeon. PMID- 9849348 TI - Laparoscopic cholecystectomy, bile duct and the British and Irish surgeon. PMID- 9849349 TI - Is the incidence of acute appendicitis really falling? PMID- 9849350 TI - What's the serum albumin? PMID- 9849352 TI - Functional food science in Europe. PMID- 9849351 TI - How long should suction drains stay in after breast surgery with axillary dissection? PMID- 9849353 TI - Growth, development and differentiation: a functional food science approach. AB - Few other aspects of food supply and metabolism are of greater biological importance than the feeding of mothers during pregnancy and lactation, and of their infants and young children. Nutritional factors during early development not only have short-term effects on growth, body composition and body functions but also exert long-term effects on health, disease and mortality risks in adulthood, as well as development of neural functions and behaviour, a phenomenon called 'metabolic programming'. The interaction of nutrients and gene expression may form the basis of many of these programming effects and needs to be investigated in more detail. The relation between availability of food ingredients and cell and tissue differentiation and its possible uses for promoting health and development requires further exploration. The course of pregnancy, childbirth and lactation as well as human milk composition and the short- and long-term outcome of the child are influenced by the intake of foods and particularly micronutrients, e.g. polyunsaturated fatty acids, Fe, Zn and I. Folic acid supplementation from before conception through the first weeks of pregnancy can markedly reduce the occurrence of severe embryonic malformations; other potential benefits of modulating nutrient supply on maternal and child health should be further evaluated. The evaluation of dietary effects on child growth requires epidemiological and field studies as well as evaluation of specific cell and tissue growth. Novel substrates, growth factors and conditionally essential nutrients (e.g. growth factors, amino acids, polyunsaturated fatty acids) may be potentially useful as ingredients in functional foods and need to be assessed carefully. Intestinal growth, maturation, and adaptation as well as long-term function may be influenced by food ingredients such as oligosaccharides, gangliosides, high-molecular-mass glycoproteins, bile salt-activated lipase, pre- and probiotics. There are indications for some beneficial effects of functional foods on the developing immune response, for example induced by antioxidant vitamins, trace elements, fatty acids, arginine, nucleotides, and altered antigen contents in infant foods. Peak bone mass at the end of adolescence can be increased by dietary means, which is expected to be of long-term importance for the prevention of osteoporosis at older ages. Future studies should be directed to the combined effects of Ca and other constituents of growing bone, such as P, Mg and Zn, as well as vitamins D and K, and the trace elements F and B. Pregnancy and the first postnatal months are critical time periods for the growth and development of the human nervous system, processes for which adequate substrate supplies are essential. Early diet seems to have long-term effects on sensory and cognitive abilities as well as behaviour. The potential beneficial effects of a balanced supply of nutrients such as I, Fe, Zn and polyunsaturated fatty acids should be further evaluated. Possible long-term effects of early exposure to tastes and flavours on later food choice preferences may have a major impact on public health and need to be further elucidated. The use of biotechnology and recombinant techniques may offer the opportunity to include various bioactive substances in special dietary products, such as human milk proteins, peptides, growth factors, which may have beneficial physiological effects, particularly in infancy and early childhood. PMID- 9849354 TI - Functional food science and substrate metabolism. AB - The present review addresses the role of food constituents in the aetiology of metabolic conditions and chronic diseases, mostly related to energy metabolism and substrate regulation, such as obesity and non-insulin-dependent diabetes mellitus. Second, attention is paid to malnutrition, a major cause of mortality and morbidity in developing countries, which may be a cause of concern in Europe because of the increasing number of elderly people in the population. Finally, the role of diet during exercise, a condition of enormous substrate demands, is evaluated. Based on a critical evaluation of the existing knowledge in the literature, implications for future research in relation to functional foods are discussed. PMID- 9849355 TI - Functional food science and defence against reactive oxidative species. AB - This paper assesses critically the science base that underpins the argument that oxidative damage is a significant causative factor in the development of human diseases and that antioxidants are capable of preventing or ameliorating these disease processes. The assessment has been carried out under a number of headings, and some recommendations for future research are made based on the present day knowledge base. The knowledge database (1) Consideration of the basic science that underlies understanding of the role of free radicals in causing cellular pathologies, and the role of antioxidants in preventing this, shows that an imbalance of reactive oxygen species and antioxidant defence systems may lead to chemical modifications of biologically relevant macromolecules. This imbalance provides a logical pathobiochemical mechanism for the initiation and development of several disease states. Experimental data obtained in vivo provide evidence that antioxidants function in systems that scavenge reactive oxygen species and that these are relevant to what occurs in vivo. The relevance in vivo of these observations depends inter alia on knowledge of the uptake and distribution of the antioxidant within the human body, and on what tissue levels of the antioxidant may be expected in relation to dietary levels. (2) There is some way to go until validated precise methods are available for measuring biomarkers of oxidative damage in human subjects in vivo under minimally invasive conditions. With respect to oxidative damage in DNa, HPLC and GC-mass spectrophotometry methods have both merits and limitations. Lipid oxidation products in plasma are best measured as isoprostanes or as lipid hydroperoxides using specific HPLC techniques. Development of isoprostane measurement will advance specificity and precision. The measurement of oxidative damage to proteins has some potential but such methods have not been effectively exploited. (3) Epidemiological studies support the hypothesis that the major antioxidant nutrients vitamin E and vitamin C, and beta-carotene (which may or may not be acting as an antioxidant in vivo), may play a beneficial role in prevention of several chronic disorders. More research is needed on the impact of other non-nutrient compounds, such as other carotenoids and flavonoids, on human health. In general, human intervention studies using hard end-points are the gold standard. Trials are restricted mainly to the major antioxidants and do not allow firm conclusions because of inconsistent findings, an insufficient number of studies and the use of varying doses. There is evidence that large doses of beta-carotene may be deleterious to the health of certain subgroups of the population such as heavy habitual smokers. (4) With respect to the safety of administration of supplementary vitamins, vitamin C is safe at levels of supplementation up to 600 mg/d, and higher levels, up to 2000 mg/d, are without risk. Vitamin E has a very low human toxicity and an intake of 1000 mg/d is without risk; 3200 mg/d has been shown to be without any consistent risk. Large intakes of beta-carotene must be viewed with caution because they have been shown to confer detriment to a population at high risk of lung cancer when administered after many years of high risk (smoking) behaviour. Until further work clarifies the situation in heavy smokers with respect to taking supplements, larger doses should be avoided by such individuals. There is little reliable information about the human toxicology of flavonoids and related non-nutrient antioxidant constituents of the diet. (5) The food industry has long experience in the control of oxidative damage in foods and this experience can be used to advantage for the protection of food antioxidants which are beneficial. Some of these, such as vitamins C and E and beta-carotene, are well known, and strategies for their protection in foods are already exploited by food technologies. (ABSTRACT TRUNCATED) PMID- 9849356 TI - Functional food science and the cardiovascular system. AB - Cardiovascular disease has a multifactorial aetiology, as is illustrated by the existence of numerous risk indicators, many of which can be influenced by dietary means. It should be recalled, however, that only after a cause-and-effect relationship has been established between the disease and a given risk indicator (called a risk factor in that case), can modifying this factor be expected to affect disease morbidity and mortality. In this paper, effects of diet on cardiovascular risk are reviewed, with special emphasis on modification of the plasma lipoprotein profile and of hypertension. In addition, dietary influences on arterial thrombotic processes, immunological interactions, insulin resistance and hyperhomocysteinaemia are discussed. Dietary lipids are able to affect lipoprotein metabolism in a significant way, thereby modifying the risk of cardiovascular disease. However, more research is required concerning the possible interactions between the various dietary fatty acids, and between fatty acids and dietary cholesterol. In addition, more studies are needed with respect to the possible importance of the postprandial state. Although in the aetiology of hypertension the genetic component is definitely stronger than environmental factors, some benefit in terms of the development and coronary complications of atherosclerosis in hypertensive patients can be expected from fatty acids such as alpha-linolenic acid, eicosapentaenoic acid and docosahexaenoic acid. This particularly holds for those subjects where the hypertensive mechanism involves the formation of thromboxane A2 and/or alpha 1-adrenergic activities. However, large-scale trials are required to test this contention. Certain aspects of blood platelet function, blood coagulability, and fibrinolytic activity are associated with cardiovascular risk, but causality has been insufficiently proven. Nonetheless, well-designed intervention studies should be initiated to further evaluate such promising dietary components as the various n-3 and n-6 fatty acids and their combination, antioxidants, fibre, etc. for their effect on processes participating in arterial thrombus formation. Long-chain polyenes of the n-3 family and antioxidants can modify the activity of immunocompetent cells, but we are at an early stage of examining the role of immune function on the development of atherosclerotic plaques. Actually, there is little, if any, evidence that dietary modulation of immune system responses of cells participating in atherogenesis exerts beneficial effects. Although it seems feasible to modulate insulin sensitivity and subsequent cardiovascular risk factors by decreasing the total amount of dietary fat and increasing the proportion of polyunsaturated fatty acids, additional studies on the efficacy of specific fatty acids, dietary fibre, and low-energy diets, as well as on the mechanisms involved are required to understand the real function of these dietary components. Finally, dietary supplements containing folate and vitamins B6 and/or B12 should be tested for their potential to reduce cardiovascular risk by lowering the plasma level of homocysteine. PMID- 9849357 TI - Functional food science and gastrointestinal physiology and function. AB - The gut is an obvious target for the development of functional foods, acting as it does as the interface between diet and the metabolic events which sustain life. The key processes in digestive physiology which can be regulated by modifying diet are satiety, the rate and extent of macronutrient breakdown and absorption from the small bowel, sterol metabolism, the colonic microflora, fermentation, mucosal function and bowel habit, and the gut immune system. The intestinal microflora is the main focus of many current functional foods. Probiotics are foods which contain live bacteria which are beneficial to health whilst prebiotics, such as certain non-digestible oligosaccharides which selectively stimulate the growth of bifidobacteria in the colon, are already on the market. Their claimed benefits are to alleviate lactose maldigestion, increase resistance to invasion by pathogenic species of bacteria in the gut, stimulate the immune system and possibly protect against cancer. There are very few reports of well-designed human intervention studies with prebiotics as yet. Certain probiotic species have been shown to shorten the duration of rotavirus diarrhoea in children but much more work is needed on the mechanism of immunomodulation and of competitive exclusion and microflora modification. The development of functional foods for the gut is in its infancy and will be successful only if more fundamental research is done on digestive physiology, the gut microflora, immune system and mucosal function. PMID- 9849358 TI - Functional food science and behaviour and psychological functions. AB - The impact of ingesting various foods on psychological and behavioural functions is a topic of both interest and concern to the general public. In this article, the scientific literature concerning demonstrated cause-and-effect relationships is reviewed, beginning with methodological considerations specific to the quantification of particular behaviours and psychological events. The essential function of food is to satisfy hunger and the need for essential nutrients. The contributions of macronutrients to appetite and satiety are described, as well as their impact on metabolism and energy balance. Functional properties of macronutrient substitutes (high-intensity sweeteners, fat replacers) and flavour enhancers are examined in relation to their contribution to hunger, satiety, and energy balance. The effects of foods and individual nutrients on the performance of diverse psychomotor tasks are studied with consideration given to the various validated quantitative tools used to assess behaviour. The effects of food components on activation, sedation, and affective states such as dysphoria are also reviewed, with special attention given to brain function and neuroactive substances such as serotonin and the endorphins. The case of hyperactivity in children is given special emphasis with reference to the potential influence of sugar and food additives. Safety issues related to food constituents and additives are discussed. Finally, a set of criteria is proposed for the evaluation and elaboration of studies in the behavioural and psychological fields, along with suggestions for future research. PMID- 9849359 TI - The Medical Devices Agency. PMID- 9849360 TI - Sir Harold Gillies Memorial Lecture. Aesthetic plastic surgery and the future plastic surgeon. PMID- 9849361 TI - Expandable anatomical implants in breast reconstructions: a prospective study. AB - Following success with two-stage anatomical breast reconstructions, we started using expandable anatomical implants for breast reconstruction to achieve one stage reconstruction. Sixty-three breast reconstructions were carried out with one-stage expandable anatomical implants over a period of 15 months, September 1995 to December 1996. To assess the performance of these implants, 20 patients with mean age 51.1 years (range 34-68 years), who were operated between February 1996 and June 1996, were entered into a prospective study. Overall, very satisfactory results were achieved, with a mean follow-up of 10.1 months (range 8 12 months). Minor complications were recorded in 3 patients and minor secondary adjustment was required in 7 patients. PMID- 9849362 TI - Perforator based V-Y advancement flaps in the leg. AB - We present a selected group of patients (18) who underwent excision of various malignant skin lesions in the leg, and had the defect resurfaced with V-Y advancement flaps. The mobility and reliability of this type of flap was enhanced by raising it as a fasciocutaneous flap based on one or two leg perforators. As patients were mobilised as soon as they recovered from the operation, there was minimal postoperative morbidity. This type of flap has the added advantage of leaving no significant donor defect and therefore better cosmesis. PMID- 9849363 TI - Contribution of collateral sprouting to the sensory and sudomotor recovery in the human palm after peripheral nerve injury. AB - The contribution of collateral sprouting to the sensory and sudomotor recovery was studied in 52 patients aged 3-66 years (mean 35.5 years) from 2 to 9 years following nerve injury and repair. The study included three groups of patients: (1) patients with complete division of median and ulnar nerves (skin reinnervation exclusively due to axon regeneration), (2) patients with isolated division of ulnar or median nerve (skin reinnervation due to axon regeneration and possible collateral sprouting), and (3) patients in whom injured axons failed to regenerate (skin reinnervation exclusively due to collateral sprouting). The end stage of sensory and sudomotor recovery was studied by: clinical methods, sensory nerve action potential (SNAP) measurements, sympathetic skin response (SSR) and the ninhydrin test. We found that recovery of sensory and sudomotor function in groups 1 and 2 was similar. End-stage sudomotor and sensory recovery within the autonomous area of the nerve did not depend on possible collateral reinnervation. Collateral reinnervation from the uninjured nerve was limited to the border innervation area of the palm and ring finger. Adjacent uninjured nerve may contribute to sprouting of nociceptive axons providing a protective function. PMID- 9849364 TI - A comparison of the patient and surgeon opinion on the long-term aesthetic outcome of reduction mammaplasty. AB - The aim of this study was to assess the difference in opinion between patients and surgeons regarding the aesthetic outcome of reduction mammaplasty. A total of 34 women, who were more than 1 year post surgery, attended an outpatient clinic to assess their opinion of the aesthetic outcome of their breast reduction. A questionnaire was used to standardise their responses. Photographic slides were taken to record the frontal, left oblique and recumbent view of their torso. These slides were assessed by four consultant plastic surgeons who completed the same questionnaire, and were blinded as to the surgeon and patient. The majority of patients rated the aesthetic outcomes of their surgery significantly higher than the consultants. Scarring was the most frequent cause of dissatisfaction for both surgeons and patients. The consultants considered the scarring following Lejour reduction to be significantly better than that following the inferior mound reduction. The nipple was considered to be too high on the breast by 12% of women but they did not request correction of this. However, consultants thought this was a problem in 27% of cases. The aesthetic outcome of reduction mammaplasty was acceptable to the patients although surgical assessment indicates that there is scope for improvement. The main area of aesthetic dissatisfaction remains the postoperative scarring. PMID- 9849365 TI - Defibrotide activity in experimental frostbite injury. AB - The pathogenesis of frostbite injury has not been completely elucidated although the available evidence suggests it is an inflammatory reaction following reperfusion injury. Defibrotide given i.p. at 40 mg/kg/ day for three days to rabbits, the ears of which were subjected to frostbite, decreased the presence of inflammatory cells (mast cells -76%; neutrophils -40.4%) and increased prostaglandin I2 (PGI2) (as 6-Keto-PGF1 alpha) in the involved skin. Thromboxane A2 (TxA2) (as TxB2) was unaffected. These data strengthen the view that an inflammatory process is the underlying cause of frostbite injury and that Defibrotide is active in pathological situations involving an inflammatory process like in frostbite. PMID- 9849366 TI - The origin and mode of fibroblast migration and proliferation in granulation tissue. AB - The factors which regulate the properties of granulation tissue remain uncertain. The key cells in granulation tissue are fibroblasts and their role was investigated in rat skin excisional wounds. Full thickness wounds in 28 Hooded Lister rats were made and traced. On days 1 to 7, animals in groups of 4 received colchicine 1 mg/kg subcutaneously, 4 h before being killed. Wound sections were stained to locate and count mitotic cells. All wounds healed with a normal coefficient of contraction. The fibroblasts' mitotic indices (% of mitotic cells) were calculated. Dermis: 0.1% vs. 2.9% (control vs. test P < 0.001, paired t test); fascia; 0.07% vs. 5.6% (control vs. test P < 0.001); granulation tissue: 9.7%. Granulation tissue fibroblasts come mainly from the adjacent fascia although the dermis may have an initial role. Once they are in the wound replication is extremely rapid. This study highlights for the first time the rapidity of fibroblastic migration and proliferation in excisional wounds, and suggests that future therapeutic interventions to control granulation tissue should consider these kinetics. PMID- 9849367 TI - Topical tamoxifen--a potential therapeutic regime in treating excessive dermal scarring? AB - Abnormal dermal scarring which affects a large number of people is aesthetically disfiguring and can be functionally disabling. Existing medical and surgical strategies to prevent or to treat scars are frequently disappointing and more effective therapies are needed. Tamoxifen, which has been used extensively in the treatment of breast cancer over the last 20 years has recently been shown to inhibit the proliferation of fibroblasts cultured from keloid biopsies. Successful treatment of retroperitoneal fibrosis and desmoid tumours with tamoxifen has also been reported. We have investigated the potential of tamoxifen as an inhibitor of wound contraction, using fibroblast-populated collagen lattices as an in vitro model. From these studies we postulate that tamoxifen may have potential clinical significance in the treatment of abnormal scarring. Normal adult human skin fibroblasts were embedded within type I collagen, then medium either with or without addition of tamoxifen was added to the collagen lattices. Lattice diameters were measured at intervals to assess the influence of tamoxifen on the lattice contraction. The reversibility of the inhibitory effect of tamoxifen on lattice contraction was investigated by 'washing out the tamoxifen' at different time-points. To visualise changes in the morphology of fibroblasts MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] was added to the lattices. Tamoxifen at 1 and 5 microM had no significant influence on lattice contraction but higher concentrations of 50 and 100 microM completely inhibited contraction. At intermediate concentrations from 10 to 20 microM the degree of lattice contraction was dose-dependent. The reversibility of the inhibition was both dose- and time-dependent. Both the inhibition of contraction and the reversibility of inhibition appeared to correlate with changes in fibroblast morphology. The dose- and time-dependent inhibition of contraction by fibroblasts suggests that tamoxifen could be investigated as a novel potential therapeutic agent in treating abnormal dermal scarring. PMID- 9849368 TI - Salvage of a below knee amputation stump with a free sensate total sole flap preserving continuity of the posterior tibial nerve. AB - We illustrate the use of a free sole flap with intact posterior tibial nerve in the coverage of a below knee amputation 6 months after an explosive injury to a 12-year-old child. We discuss the indications for lower leg amputation in children, modalities of reconstruction of the amputation stump and possible reason for the observed change of colour of the flap persisting for 14 days. PMID- 9849369 TI - Heel replantation. AB - A 14-year-old boy presented with oblique amputation of the heel through the calcaneum. The heel fragment was successfully replanted but the postoperative course was complicated by venous insufficiency requiring further venous anastomosis and attributed to tension in the skin closure. PMID- 9849370 TI - Cone-shaped epiphyses in the toes and trifurcation of the soft palate in oral facial-digital syndrome type-I. AB - A rare case of oral-facial-digital syndrome type I is presented. The patient had two features that have not been previously described: cone-shaped epiphyses in the toes and trifurcation of the soft palate. PMID- 9849371 TI - Oromandibular-limb hypogenesis syndromes: a case of aglossia with an intraoral band. AB - Oromandibular and limb syndromes feature primarily in sporadic case reports because of their low incidence. They include Moebius syndrome, aglossia-adactylia syndrome, Hanhart syndrome, glossopalatine ankylosis syndrome, limb deficiency splenogonadal fusion syndrome and Charlie M. syndrome. There is confusion in the classification of these patients because of the associated anomalies and the frequency of overlapping features. This paper presents a patient with oromandibular malformations associated with major defects in the upper and lower limbs. Aglossia in the presence of an intraoral band is a peculiar association demanding classification. This case confirms that aglossia-adactylia syndrome and the glossopalatine ankylosis syndrome are variants along a spectrum. PMID- 9849372 TI - Endoscopic join the dots! PMID- 9849373 TI - Incremental increases in Breslow thickness of thick melanoma have a linear effect on mortality. PMID- 9849374 TI - Subscapular vascular axis for raising flaps. PMID- 9849375 TI - Accidental burns following subcutaneous mastectomy and reconstruction with a prosthesis. PMID- 9849376 TI - Cutting diathermy for removal of implants. PMID- 9849377 TI - Non-Hodgkins lymphoma masquerading as a chronic venous leg ulcer. PMID- 9849378 TI - Verification systems--tools or toys? PMID- 9849379 TI - Patient doses in spiral CT. PMID- 9849380 TI - Target volume definition for three-dimensional conformal radiation therapy of lung cancer. AB - Three-dimensional conformal radiation therapy (3DCRT) is a mode of high precision radiotherapy which has the potential to improve the therapeutic ratio of radiation therapy for locally advanced non-small cell lung cancer. The preliminary clinical experience with 3DCRT has been promising and justifies further endeavour to refine its clinical application and ultimately test its role in randomized trials. There are several steps to be taken before 3DCRT evolves into an effective single modality for the treatment of lung cancer and before it is effectively integrated with chemotherapy. This article addresses core issues in the process of target volume definition for the application of 3DCRT technology to lung cancer. The International Commission on Radiation Units and Measurements Report no. 50 definitions of target volumes are used to identify the factors influencing target volumes in lung cancer. The rationale for applying 3DCRT to lung cancer is based on the frequency of failure to eradicate gross tumour with conventional approaches. It may therefore be appropriate to ignore subclinical or microscopic extensions when designing a clinical target volume, thereby restricting target volume size and allowing dose escalation. When the clinical target volume is expanded to a planning target volume, an optimized margin would result in homogeneous irradiation to the highest dose feasible within normal tissue constraints. To arrive at such optimized margins, multiple factors, including data acquisition, data transfer, patient movement, treatment reproducibility, and internal organ and target volume motion, must be considered. These factors may vary significantly depending on technology and techniques, and published quantitative analyses are no substitute for meticulous attention to detail and audit of performance. PMID- 9849381 TI - Ultrasound and Doppler features of accessory spleens and splenic grafts. AB - 16 splenic grafts and 24 accessory spleens were evaluated with ultrasound, colour Doppler and power Doppler imaging, in order to compare the features of splenic grafts and accessory spleens. 12 splenic grafts (11 surgically implanted, 1 islet of splenosis) were detected in eight patients following splenectomy 6-12 months previously. Four of the surgically implanted splenic grafts were found in three patients following splenectomy 16-18 days previously. 21 accessory spleens were incidentally detected in 20 subjects who had not been operated upon, whereas three enlarged accessory spleens were found in 10 patients who underwent splenectomy 6-12 months previously. The size, shape, rim features, echogenicity, acoustic enhancement and vascularization of the splenic grafts and accessory spleens were evaluated. The splenic grafts were oval with lobulated margins, an inhomogeneous pattern in 36% and acoustic enhancement in 64%. Multiple feeding vessels arising from the surrounding tissue entered the periphery of 10 splenic grafts in the eight patients operated on 6-12 months previously, whereas no vascular signals were detected in the four splenic grafts of the three patients operated on 16-18 days ago. Accessory spleens were round with smooth echogenic margins and a vascular hilum arising from the splenic vessels. In conclusion colour Doppler and power Doppler allow differentiation between accessory spleens and splenic grafts basing on their shape and vascular supply. PMID- 9849382 TI - CT angiography in vascular intervention for steno-occlusive diseases: role of multiplanar reconstruction and source images. AB - The aim of this study was to assess the efficacy of CT angiography for steno occlusive diseases before and after interventional procedures, focusing on the role of multiplanar reconstruction (MPR) and source images. 17 patients with 20 steno-occlusive lesions underwent CT angiography before and after interventional procedures. For each lesion, the percentage stenosis obtained on CT angiography was compared with that on conventional angiography. In addition, MPR and source images were evaluated for the presence of wall thickening and calcification before interventional procedures, and the presence of dissection and luminal shape after interventional procedures. These findings were compared with those of conventional angiography. Although the percentage stenosis depicted on CT angiography correlated well with that on conventional angiography, MPR and source images clearly demonstrated the effect of intervention and the residual stenosis. MPR and source images clearly depicted wall thickening, wall calcification, the presence of dissection and the luminal shape. CT angiography provides useful information before intervention, while MPR and source images are of value in evaluating arterial wall abnormalities and morphological changes associated with interventional procedures. PMID- 9849383 TI - Evaluation of cystic ovarian lesions using apparent diffusion coefficient calculated from turboFLASH MR images. AB - Serial turboFLASH (fast low-angle shot) images with and without diffusion perfusion (DP) gradients were used to evaluate the contents of cystic ovarian lesions. 19 ovarian cysts seven serous cystadenomas, six mucinous cystadenomas and 13 malignant cystic ovarian tumours were studied. T1 and T2 weighted spin echo images, serial turboFLASH images with and without DP gradients (b = 295.1 and 0 s mm-2, respectively) were obtained. Using these images, the apparent diffusion coefficients (ADCs) were calculated within the cystic contents of these lesions. Approximately corrected ADC (ADC') values were also calculated in order to reduce T1 contamination from ADCs, using results of a phantom study. Ovarian cysts and serous cystadenomas showed larger ADC and ADC' values than the cystic contents of malignant ovarian lesions (p < 0.01). Mucinous cystadenomas show no distinct ADC difference to any other group of cases. However, if the highest ADC' loculus was selected as being representative, mucinous cystadenomas showed a larger ADC' value than malignant cystic neoplasms (p < 0.03). In conclusion, diffusion-weighted MRI is possibly of use in evaluating cystic contents of ovarian lesions. PMID- 9849384 TI - Percutaneous biopsy of small hepatic lesions using an 18 gauge automated needle. AB - This study evaluates the efficacy of ultrasound guided percutaneous biopsy using an 18 gauge automated side-cutting needle in the diagnosis of small (< or = 3 cm) focal hepatic lesions. 137 consecutive percutaneous biopsies of 131 different small (< or = 3 cm) focal hepatic lesions were included. 11 biopsies were performed on lesions of < or = 1 cm in diameter, 56 were on lesions 1.1-2 cm in size and 70 on lesions 2.1-3 cm in size (average 2.3 +/- 0.7 cm; median 2.3 cm; range 0.7-3 cm). The biopsy specimen was sufficient for diagnosis in 135 cases (98.5%). The sensitivity for diagnosing malignancy was 96.4%; specificity was 100%, positive and negative predictive values were 100% and 94.6%, respectively; accuracy was 97.8%. There was no statistically significant difference in the diagnostic efficacy for lesions of different pathology and size. No significant post-biopsy complication occurred. It is concluded that the 18 gauge Temno needle is safe and effective in diagnosing small hepatic lesions. PMID- 9849385 TI - Simultaneous densitometry and quantitative bone sonography in the estimation of osteoporotic fracture risk. AB - 759 post-menopausal women (41-80 years old), 175 with and 584 without vertebral fracture, were studied by quantitative bone sonography and densitometry of the distal radius in order to evaluate the ability of ultrasound transmission velocity (UTV) to separate fractured from healthy women independently of bone mineral density (BMD) and to test the possibility of improving the discriminant ability of BMD by the simultaneous use of UTV. A second BMD measurement was made at the mid radial shaft. Both BMD and UTV were higher in healthy subjects than in fractured women; the latter being older, shorter and having a longer postmenopausal status. On logistic regression, standardized for 1 SD from the mean value of healthy women, UTV differentiated between healthy and fractured subjects after correction for BMD. UTV was also a significant predictor of fracture in a selected subgroup of healthy and fractured women paired for BMD (144 pairs). In this same subgroup, the difference in UTV between fractured and healthy women regression lines was related to elasticity (E) variation between pairs. Simultaneously evaluating BMD and UTV as fracture predictors, logistic regression showed an odds ratio that was twice that of each predictor alone and 1.2 times higher than that derived from the simultaneous evaluation of two different BMD predictors. These data confirmed that UTV differentiates between healthy and fractured women both as well as and independent of BMD. In addition, UTV separated fractured from healthy BMD matched women by measuring non mass related differences. The simultaneous use of BMD and UTV improved the discriminant ability of each of them alone and of two simultaneous measurements of BMD detected at different skeletal sites. PMID- 9849386 TI - Radio-opaque markers for stereotactic imaging of uveal melanoma. AB - In in vitro and in vivo studies, episclerally sutured radio-opaque markers were evaluated as localizers for better determination of the clinical tumour volume prior to stereotactic radio-therapy of uveal melanoma. Four different types of markers were studied in vitro: tantalum marker, 2.5 mm in diameter; ring-shaped markers custom-designed in polymethyl methacrylate (PMMA), 2.6 mm and 3.0 mm in diameter; and barium-impregnated silicone rubber spheres, 2 mm in diameter. In vivo PMMA markers 3.0 mm in diameter and barium-impregnated silicone rubber spheres 2 mm in diameter were used. The best results were obtained with the barium-impregnated silicone rubber spheres both in vitro and in vivo. For the CT delineation of selected uveal melanomas with either a flat discoid shape or flat tumour extensions or when adjacent serous retinal detachment is present, small episclerally sutured barium-impregnated silicone rubber spheres are useful as localizers for determining the clinical tumour volume. PMID- 9849387 TI - Patient radiation doses during cardiac catheterization procedures. AB - The objective of the present project was the determination of the dose received by patients during cardiac procedures, such as coronary angiography, percutaneous transluminal coronary angioplasty (PTCA) and stent implantation. Thermoluminescent dosemeters (TLDs), suitably calibrated, were used for the measurement of the dose received at four anatomical locations on the patient's skin. A dose-area product (DAP) meter was also used. The contribution of cinefluorography to the total DAP was higher than that of fluoroscopy. A DAP to effective dose conversion factor equal to 0.183 mSv Gy-1 cm-2 was estimated with the help of a Rando phantom. Thus, the effective dose received by the patients could be assessed. Mean values of effective dose equal to 5.6 mSv, 6.9 mSv, 9.3 mSv, 9.0 mSv and 13.0 mSv were estimated for coronary angiography, PTCA, coronary angiography and ad hoc PTCA, PTCA followed by stent implantation and coronary angiography and ad hoc PTCA followed by stent implantation, respectively. PMID- 9849388 TI - A study of chest radiography with mobile X-ray units. AB - A survey of radiographic technique and estimated entrance surface dose has been carried out for 364 chest radiographs performed with mobile X-ray equipment in the Intensive Therapy Unit (ITU) and 30 wards at Aberdeen Royal Infirmary. Data for these two types of location were compared, as were those for two film/screen systems used on the wards. Image quality assessments were made on sets of radiographs for two patients. Entrance skin doses for chest radiographs performed in the ITU were 50% greater than on the wards with the same film/screen system. The main technique difference was the use of shorter focus-to-skin distances (FSDs) in ITU. Doses with the Kodak Insight system were 20% higher than those using Du Pont Quanta III in similar locations. No correlation was found between image quality and entrance surface dose (ESD). Results from the survey were used to recommend exposure factors for shorter FSDs. A follow-up study revealed a 35 45% reduction in ESD for Kodak Insight and a 20% reduction for Quanta III. PMID- 9849389 TI - Estimation of compressed breast thickness during mammography. AB - To estimate radiation dose during mammography the breast thickness must be known. We present a new method for estimating the thickness of a compressed breast using only the breast image as projected onto a mammogram, calibration data such as the mAs value and image processing techniques. The method proves to be of high accuracy (+/- 0.2 cm for craniocaudal mammograms) and has the advantage over other methods of allowing retrospective estimation of thickness. PMID- 9849391 TI - In vivo semiconductor dosimetry as part of routine quality assurance. AB - This paper describes the initial physics testing necessary before diodes can be used for in vivo dosimetry as well as the development of a protocol for clinical use in head and neck treatment and the preliminary results acquired. 50 patients were entered into the pilot study. A total of 300 treatment set-ups were measured (184 entrance doses and 116 exit doses). Wedged and unwedged components of each field were measured separately, making the total number of entrance doses 284 and total number of exit doses 207. There was no significant systematic deviation in the measured entrance dose compared with the expected (mean +0.4%, SD 2.7%). Discrepancies between the observed and expected entrance doses of greater than 5% were recorded in 6% (16/284) of measurements. The mean of the measured exit doses was 2.4% lower than expected (SD 4.8%). Discrepancies between the observed and expected exist doses of greater than 5% were recorded in 32% (67/207) of measurements. Possible causes for these discrepancies are discussed. Overall analysis of the data for individual patients suggest that in one patient out of the 50 there may have been a delivered target volume dose discrepancy of greater than 5% (+6.5%). The significance of the results and the implications for routine use are discussed. PMID- 9849390 TI - Doses to patients in routine X-ray examinations in Malaysia. AB - A collaborative national survey initiated by the University of Malaya and the Ministry of Health was conducted from 1993 to 1995 to establish baseline patient dose data for seven common types (12 projections) of X-ray examinations in Malaysia. A total of 12 randomly selected public hospitals and 867 patients were included in this survey. The entrance surface doses (ESD) received by the patients were measured using thermoluminescent dosemeters (TLDs) attached to the patient's skin. Histograms are presented showing wide, positively skewed distributions of measured entrance surface doses for each examination. Mean, median, first and third quartile values of ESD and median effective dose are reported. Survey results are generally comparable with those reported in the UK, USA and by the International Atomic Energy Agency (IAEA). The results also provide information on dose level for a lower weight population (mean weight 60 kg) compared with the international reference dose values based on a 70 kg standard. The findings support the importance of the on-going national quality assurance programme to ensure doses are kept to a level consistent with optimum image quality. The data will also be useful for the formulation of national guidance levels as recommended by the IAEA. Furthermore, this study provides patient dosimetry information on healthcare level II countries. PMID- 9849392 TI - Memorandum from the British Committee on Radiation Units and Measurements. RBE values for the retrospective calculation of risk to specific organs as a result of neutron irradiation. PMID- 9849393 TI - Patient and staff radiation dose in fluoroscopy-guided TIPS procedures and dose reduction, using dedicated fluoroscopy exposure settings. AB - Fluoroscopy guided interventions, such as transjugular intrahepatic portosystemic shunt (TIPS) procedures, can results in relatively high radiation doses to patients and staff. The purpose of this study was to evaluate the possible benefit of dedicated fluoroscopy exposure factors in the reduction of doses. Doses to patients and staff were measured during fluoroscopy-guided TIPS procedures in two Dutch university hospitals. Patient doses were calculated from dose-area product (DAP) measurements, entrance beam dimensions and DAP conversion factors. Staff doses were measured outside lead aprons using electronic personal dosemeters. Average patient entrance skin dose (ESD) rate during fluoroscopy was 49 mGy min-1 (13 cases, average fluoroscopy duration 32 min) in one hospital, and 6 mGy min-1 (10 cases, average fluoroscopy duration 50 min) in the other. Estimated staff effective dose per procedure was 28 microSv average in the first hospital compared with 4 microSv average in the other. The use of dedicated fluoroscopy exposure factors, with a relatively high tube voltage and lower tube current resulted in a significant dose reduction for patient and staff in this type of radiological intervention. PMID- 9849394 TI - Benign solitary fibrous tumour of the pre-sacral space: MRI findings. AB - Benign solitary fibrous tumour, a rare mesenchymal tumour of adults, usually arises from the pleura. Only a few cases have been reported in the retroperitoneum and, to our knowledge, there has been no report of its imaging features. We describe the MRI features of benign solitary fibrous tumour arising from the pre-sacral space. PMID- 9849395 TI - Tuberculous pericarditis: MRI features with contrast enhancement. AB - Tuberculous pericarditis has decreased in incidence, but early diagnosis and treatment are critical because constrictive pericarditis is a serious complication of this condition. The appearances of tuberculous pericarditis on gadolinium enhanced magnetic resonance imaging are described and the value of this modality in achieving an early diagnosis is emphasized. PMID- 9849396 TI - Mucinous biliary cystadenocarcinoma containing gas bubbles secondary to duodenal invasion. AB - Biliary cystadenocarcinoma is a rare, usually intrahepatic neoplasm. A case is described in which an intrahepatic cystadenocarcinoma invaded the duodenum. This resulted in intratumoral gas. PMID- 9849397 TI - Enhancement of subtle density differences of the lung parenchyma on CT. AB - Several pulmonary conditions are characterized by the finding of areas of decreased attenuation relative to adjacent normal lung parenchyma on computed tomography. In particular, the identification of areas of the lung affected by small airways disease is difficult as the reduction in lung density is subtle and the regions are poorly marginated. This paper presents a novel wavelet analysis technique which increases the conspicuity of differences between normal and abnormal lung parenchyma. PMID- 9849398 TI - An unusual twist. PMID- 9849399 TI - Rupture and growth of adrenal myelolipoma in two patients. PMID- 9849400 TI - Bone densitometry reference data. PMID- 9849401 TI - Incidence, risk factors and prevention of melanoma. AB - The incidence of cutaneous malignant melanoma steadily increased between 1940 and 1990 in both sexes. However, this increase appears to have peaked in females in Scotland. Excessive exposure to ultraviolet radiation among Caucasians is the main etiologic factor implicated in the incidence of melanoma. Mortality due to melanoma also increased between 1940 and 1990, but the rate of increase is less than that of the incidence of melanoma. This may be due to earlier diagnosis and treatment of melanoma as a result of public education campaigns. Independent risk factors for developing melanoma include the presence of benign melanocytic naevi (moles), the development of lentigines or freckles, three or more dysplastic naevi and a history of three or more severe sunburns that resulted in peeling or blistering. Approximately 2% of melanoma patients have a family history of the disease and research into potential melanoma susceptibility genes is ongoing. Primary prevention campaigns, initiated mainly in Australia, are aimed at encouraging sensible sun exposure. Secondary prevention campaigns are directed at preventing death from melanoma by encouraging early diagnosis and treatment. Additional prospective studies are needed to determine if the incidence of melanoma has peaked and whether the recent trends observed in females will also occur in males. PMID- 9849402 TI - Role of lymphatic mapping and sentinel node biopsy in the detection of melanoma nodal metastases. AB - The approach to the clinically negative regional lymph node basin presents a challenging problem in the clinical management of patients with early-stage melanoma (stage I and II). As a group, stage I and stage II patients generally have a good prognosis following surgical excision of the primary tumour; however, the presence of clinically undetectable nodal disease is an important prognostic factor. Traditionally, these patients are either monitored closely for development of palpable metastases at which time a therapeutic lymphadenectomy would be performed or, have been offered early elective lymphadenectomy that is performed immediately at the time of wide excision of the primary tumour. The high rate of morbidity and lack of proven survival impact for immediate elective lymphadenectomy limit its use. Lymphatic mapping and sentinel node biopsy allow a more selective approach to regional lymph node dissections. Only node-positive patients undergo lymphadenectomy, thereby avoiding the morbidity associated with the procedure for patients without nodal disease. Several studies have confirmed that lymphatic mapping identifies the lymph node most likely to contain disease and confirms the orderly progression of lymphatic metastases. Use of radioactive colloid and a hand-held gamma probe improves the localisation of the sentinel lymph node. Because adjuvant systemic therapy has been proven to prolong survival in patients with nodal involvement, lymphatic mapping and sentinel node biopsy, by identifying patients with clinically negative, but pathologically positive disease, may improve their outcome. PMID- 9849403 TI - Systemic adjuvant treatment of high-risk melanoma: the role of interferon alfa-2b and other immunotherapies. AB - Until recently, the prognosis of patients with deep primary melanomas or regionally metastatic nodal disease has been poor, with 5-year survival rates of 25-50%. The results of the Eastern Cooperative Oncology Group (ECOG) trial 1684 represent the first evidence of effective adjuvant therapy for these patients. Interferon alfa-2b (IFN-alpha 2b) administered at maximally tolerated doses for 1 year significantly improved both relapse-free and overall survival. The impact of interferon therapy was observed early during treatment and the effect was durable. The results of this trial represent a breakthrough in the treatment of high-risk resected cutaneous melanoma and identify the new reference standard for new cytokines, vaccines and combinations. The favourable results provide a strong impetus for redoubled research into immunotherapy for treatment of melanoma. Specifically, ganglioside vaccines have been identified that induce antibody responses and may affect patient outcome and peptide/protein vaccines that are recognised by the T-cell have been identified in large numbers. ECOG and the U.S. Intergroup are conducting a phase III trial (E1694) that compares GM2 vaccine to IFN-alpha 2b and a phase II trial evaluating concurrent or sequential use of interferon and vaccines for patients with resectable melanoma. They are also planning phase II trials of peptides for patients with metastatic unresectable melanoma. Laboratory analyses of the immune responses induced by IFN and the several vaccines are anticipated to reveal the fundamental immune mechanisms that are important for relapse-free survival and immunological control of melanoma. PMID- 9849404 TI - Cost-effectiveness assessment of interferon alfa-2b as adjuvant therapy of high risk resected cutaneous melanoma. AB - The use of interferon alfa-2b (IFN-alpha 2b) as adjuvant therapy of high-risk resected cutaneous melanoma was recently found to significantly improve relapse free and overall survival in the Eastern Cooperative Oncology Group trial 1684 (E1684). However, treatment toxicities and costs may limit its widespread use. A cost-effectiveness and cost-utility analysis of this therapy was conducted using a hypothetical cohort of patients as if they had entered E1684. Survival and recurrence rates were calculated at 7 and 35 years for typical 50-year-old melanoma patients based on the clinical results of E1684 and natural history databases. Costs included all treatment-related costs (i.e. drug acquisition and administration, monitoring and treatment-related toxicity) and the costs of treating recurrences. Estimated utility values were assigned based on data from other oncology trials. The model predicted that IFN-alpha 2b provided an extra 0.52 years of life compared with observation at 7 years; however, at 35 years, the survival benefit of IFN-alpha 2b increased almost 4-fold to nearly 2 years. At 7 years, the cost per year of life gained was U.S. $32,600 and the cost per quality-adjusted life-year (QALY) gained was U.S. $43,200. At 35 years, these costs decreased to U.S. $13,700 and $15,200, respectively. These costs are comparable with those of other well-established medical interventions. Although these results require confirmation in a prospective study, it appears that the use of high-dose IFN-alpha 2b for patients with high-risk melanoma is cost effective. PMID- 9849405 TI - Strategy of the EORTC-MCG trial programme for adjuvant treatment of moderate-risk and high-risk melanoma. AB - Trials in high-risk melanoma patients evaluating the role of postoperative adjuvant treatment with interferons or other cytokines, ganglioside-based vaccines, or vaccines based on melanoma cells are ongoing or planned internationally. In Europe, the two largest randomised trials are carried out by the European Organization for Research and Treatment of Cancer-Melanoma Cooperative Group (EORTC-MCG). In stage IIA patients (T3N0M0) with a moderate risk of micrometastatic disease (35-40%), Trial 18,961 compares observation with ganglioside GM2 vaccination. This trial will be activated during the spring of 1998 and is expected to enrol 1000 patients. In stage IIB-IIIB (T4N0M0-TxN12M0) patients with a high risk of micrometastatic disease (approximately 80%), trial 18,952 compares observation with adjuvant therapy using two intermediate dosage regimens of interferon alfa-2b (IFN-alpha 2b). These trials and the philosophy of the EORTC-MCG programme allow more toxic treatment regimens to be investigated in patients with high-risk disease and only treatments with minimal toxicity to be evaluated in patients with moderate- to low-risk disease. Recently completed and other ongoing trials also are discussed. Overall, if clinical efficacy is demonstrated, the toxicity, impact on quality of life and treatment costs determine the acceptance and applicability of a treatment. PMID- 9849406 TI - The treatment of metastatic uveal melanoma. AB - Malignant melanoma can occur in various ocular structures and accounts for 70% of all primary eye malignancies. At the time of initial diagnosis of uveal melanoma, most patients have no demonstrable evidence of metastatic disease. However, within 5 years, metastases appear in 19-35% of patients. The median relapse-free interval for these patients is 2-4 years. Unlike cutaneous melanoma, uveal melanoma most commonly metastasises to the liver. In fact, the liver is the sole site or the initial site of metastasis in more than 50% of patients. This paper reviews major studies that evaluated different treatment options for metastatic uveal melanoma, including surgery, systemic chemotherapy, intra-arterial chemotherapy, chemoembolisation and chemoimmunotherapy. Surgery and systemic chemotherapy offer minimal benefits for these patients. However, the use of intra arterial fotemustine demonstrated a 40% response rate for patients with liver metastases of uveal melanoma. Additionally, chemoimmunotherapy with a four-drug chemotherapy regimen and interferon alfa has provided response rates of approximately 20% and may contribute to prolonged survival. Ongoing multicentre trials are expected to determine the feasibility of this regimen. The activity of immunotherapy, particularly interferon, encourages its use in combination with other active therapies, such as intra-arterial fotemustine. PMID- 9849407 TI - The evidence for tamoxifen and chemotherapy as treatment for metastatic melanoma. AB - Tamoxifen, an oestrogen antagonist routinely used in the treatment of breast cancer, has been used in clinical trials for patients with melanoma since the late 1970s. Following initial promise as a single agent for the treatment of metastatic melanoma, tamoxifen was first combined with chemotherapy in this setting in 1984. Since then, numerous phase II studies have combined tamoxifen with different chemotherapeutic agents, with some suggesting that tamoxifen significantly improves the efficacy of cisplatin-containing regimens. Overall response rates range from 8 to 60%. Several randomised trials also have been completed, with response rates of 12-30%. One such study showed statistically significant improvements in response rate and survival when tamoxifen was added to dacarbazine; however, other studies have not observed these benefits with the addition of tamoxifen to cisplatin-containing regimens. At present, the author's opinion is that the strength of evidence does not support the use of tamoxifen in combination with cisplatin-containing chemotherapy for the treatment of metastatic melanoma. Controversy remains as to whether the strength of evidence from the randomised trials outweighs the combined evidence from numerous nonrandomised trials. Resolution of this controversy may depend on the results of the North Central Cancer Therapy Group and/or a common agreement as to relative strength of evidence from clinical trials of different designs. PMID- 9849408 TI - New prognostic factors in melanoma: mRNA tumour markers. AB - Circulating tumour cells in the peripheral blood may be important for haematogenous spread of malignant disease. Monitoring these cells may therefore be of prognostic value. Reverse transcriptase-polymerase chain reaction (RT-PCR) based assays to detect occult neoplastic cells offer the highest sensitivity for the study of tumour dissemination and minimal residual disease. This review summarises technical considerations and clinical investigations in melanoma patients of various disease stages. The clinical data are promising, but to clearly define the clinical usefulness of messenger RNA (mRNA) tumour markers, methodological issues must be resolved and the clinical value must be assessed prospectively in sufficiently large patient cohorts. PMID- 9849409 TI - Future perspectives in specific immunotherapy of melanoma. AB - With the discovery of T-cell recognised tumour-associated antigens (TAAs), interest in specific immunotherapy for treatment of malignancies has increased substantially. The majority of studies investigating TAAs have focused on melanoma-associated antigens because of evidence that the immune system influences the pathogenesis of melanoma. This paper reviews the different types of melanoma antigens, their in vitro and in vivo immunogenicity and clinical data regarding the use of specific immunotherapy in patients with stage I-IV melanoma. Results of clinical studies are highly variable but encourage further research in these patients. Developing and perfecting laboratory and clinical correlates of response to these specific immunotherapies are vital to determining their role in clinical practice. PMID- 9849410 TI - Alcohol and breast cancer. PMID- 9849411 TI - Is pre-operative radiotherapy superior to postoperative radiotherapy in the treatment of soft tissue sarcoma? PMID- 9849412 TI - Methodological and statistical issues of quality of life (QoL) and economic evaluation in cancer clinical trials: report of a workshop. AB - In recent years, quality of life (QoL) and economic evaluations have become increasingly important as additional outcome measures in cancer clinical trials. However, both fields of research are relatively new and in need of finding solutions to a substantial number of specific methodological problems. This paper reports on the proceedings of a symposium aimed at summarising and discussing some of the most contentious methodological and statistical issues in QoL and economic evaluations. In addition, possible solutions are indicated and the most pertinent areas of research are identified. Issues specific to QoL evaluations that are addressed include clinically meaningful changes in QoL scores; how to analyse QoL data and to handle missing and censored data and integration of length of life and QoL outcomes. Issues specific to economic evaluations are the advantages and disadvantages of various outcome measures; statistical methods to analyse economic data and choice of decision criteria and analytical perspective. How to perform QoL and economic evaluations in large and simple trials and whether the gap between QoL and utility measures can be bridged are also discussed. PMID- 9849413 TI - Quality control of immunohistochemical evaluation of tumour-associated plasminogen activators and related components. European BIOMED-1 Concerted Action on Clinical Relevance of Proteases in Tumour Invasion and Metastasis. AB - The plasminogen activation (PA) system is involved in the breakdown and remodelling of the extracellular matrix. In the case of cancer, this is a prerequisite for invasion and metastasis. The expression of urokinase-type plasminogen activator and plasminogen activator inhibitor type 1 in particular have been reported to be of clinical and prognostic value. This has primarily been proven in the case of breast carcinoma and colon carcinoma, using the enzyme linked immunosorbent assay (ELISA) as a quantitative assay to determine the level of expression. Immunohistochemistry is another technique to investigate the presence of PA components. It allows assessment in a semiquantitative way and informs in addition on the specific distribution within the tissue. To take full advantage of the benefits of immunohistochemistry, it is important to aim at optimal quality in all steps influencing the final judgement of the staining results. These various steps are highlighted and discussed in this paper. PMID- 9849414 TI - Male breast cancer: a review of the literature. AB - Although breast cancer is uncommon in men, it can cause significant morbidity and mortality. The current review was undertaken to determine whether strategies applied for the evaluation and treatment of breast cancer in females are appropriate in male breast cancer. Male breast cancer has biological differences compared with female breast cancer, including a high prevalence in certain parts of Africa, a higher incidence of oestrogen receptor positivity and more aggressive clinical behaviour. It responds to hormonal manipulation and chemotherapy, but optimal treatment regimens in males are unknown. Male breast cancer remains an uncommon disease. Most of our current knowledge regarding its biology, natural history and treatment strategies has been extrapolated from its female counterpart. Much research is needed to further characterise the molecular biological properties of male breast tumours and their prognostic significance, and to devise treatment strategies, including optimal chemotherapy regimens. PMID- 9849415 TI - The effect of oral contraceptive use on the prognosis of node positive breast cancer patients. German Breast Cancer Study Group. AB - The effect of oral contraceptive (OC) use as a risk factor for breast cancer was recently assessed in a large meta-analysis, but currently available data on the prognostic effect are still insufficient. We investigated the relationship between OC use and standard prognostic factors and the effect of OC use on recurrence-free survival (RFS) and overall survival (OS) in 422 premenopausal pT1a-3aN + M0 patients from two trials of the German Breast Cancer Study Group (GBSG). 137 patients (32.5%) were OC users. They were younger on average (mean age 41.5 years versus 45 years for non-OC users) and the percentage of patients with smaller tumours was higher in the group of OC users. Based on 163 events for RFS and 103 events for OS, no significant effect of OC use on RFS and OS could be demonstrated in univariate and multivariate analyses. In our study of node positive breast cancer cases, OC users were younger and had smaller tumours. This may be an effect of earlier detection of breast cancer, but OC users did not have a better prognosis, both before and after adjustment for tumour size and other prognostic factors. PMID- 9849416 TI - Immunohistochemically detected p53 and P-glycoprotein predict the response to chemotherapy in lung cancer. AB - While resistance to chemotherapy is a major problem in lung cancer treatment, there is no useful predictor of treatment response. We thus designed this study to determine the utility of p53 and P-glycoprotein expression in predicting the response to chemotherapy in patients with primary lung cancer, retrospectively. We evaluated transbronchial biopsy (TBB) specimens from 60 patients with lung cancer, who were previously untreated. Formalin-fixed, paraffin-embedded TBB specimens were immunostained using anti-p53 antibody (DO-1) and anti-P glycoprotein antibody (JSB-1). The positivity of p53 was 63%, and that of P glycoprotein was 17%. No correlation was observed between p53 and P-glycoprotein immunostaining. Positivity of p53 correlated significantly (P = 0.004) with a lack of response to chemotherapy in non-small cell lung cancer (NSCLC), but not in small cell lung cancer (SCLC). In contrast, positivity of P-glycoprotein was correlated with chemotherapy resistance in SCLC (P = 0.003), but not in NSCLC. Multiple logistic regression analysis revealed that positive immunostaining for p53 was a significant risk factor for chemotherapy resistance in NSCLC. These results suggest that immunostaining of p53 and P-glycoprotein for TBB specimens may help to predict response to chemotherapy in NSCLC and SCLC, although the results should be confirmed in a larger, more homogeneous series. PMID- 9849417 TI - Phase II trial of topotecan as a 21-day continuous infusion in patients with advanced or metastatic adenocarcinoma of the pancreas. AB - The aim of this study was to determine the efficacy and toxicity of topotecan administered as a 21-day continuous intravenous infusion in patients with advanced or metastatic adenocarcinoma of the pancreas. 26 previously untreated patients with advanced or metastatic pancreatic adenocarcinoma received topotecan at a dose of 0.5 mg/m2/day or 0.6 mg/m2/day as a continuous intravenous infusion for 21 days. Courses were repeated every 28 days. 26 patients were assessable for response and toxicity on an intent-to-treat basis. The initial 8 patients at a starting dose of 0.6 mg/m2/day experienced unacceptable myelosuppression and dose delays. The subsequent 18 patients, therefore began therapy at a dose of 0.5 mg/m2/day. The major toxicity of topotecan at this dose and schedule was myelosuppression, which was reversible and non-cumulative. There were no complete responses and two partial responses for a total response rate of 8% (95% confidence interval, 1-25%). Response durations were 17 and 45 weeks. Stable disease was seen in 3 patients. The median time to progression for all patients was 8 weeks and the median survival was 20 weeks. Topotecan given as a 21-day continuous intravenous infusion has a similar response rate and median survival to our previously reported study of the 5-day short infusion regimen in pancreatic carcinoma. PMID- 9849418 TI - Bilateral testicular germ cell tumours in patients with initial stage I disease: prevalence and prognosis--a single centre's 30 years' experience. AB - Development of second testicular tumours, i.e. bilateral testicular cancer, is influenced by systemic chemotherapy for the first tumour. The prevalence of bilateral testicular cancer was studied in patients with initial stage I disease, in which no systemic treatment was given after orchidectomy. All stage I testicular cancer patients entered a surveillance study with an intensive follow up since 1982. We hypothesised that after 1982, bilateral testicular cancer was diagnosed at an earlier stage of disease. The prevalence of bilateral testicular cancer was 4.7% (8/170) in stage I patients treated between 1967 and 1981, and 2.9% (8/275) in stage I patients treated between 1982 and 1997 (P > 0.5 chi 2 test). In the period 1967-1981, 6 patients had stage I second tumours and 2 patients had stage III second tumours. The former 6 patients are alive with no evidence of disease and the 2 patients with metastatic tumours died of disease or treatment. In the period 1982-1977, all 8 patients had stage I second tumours and all are alive with no evidence of disease. The overall prevalence of bilateral testicular cancer in stage I patients was 3.6% and has slightly decreased over the past three decades. Intensive follow-up, improvement of radiodiagnostic computed tomography techniques, availability of serum tumour markers, and patient education have resulted in earlier diagnosis and lower stage of contralateral testicular tumours, contributing to improved prognosis. PMID- 9849419 TI - Dacarbazine-vindesine versus dacarbazine-vindesine-cisplatin in disseminated malignant melanoma. A randomised phase III trial. AB - In a multicentre phase III study of disseminated malignant melanoma performed in Sweden and Norway, 326 patients were randomised to receive treatment with the combination dacarbazine [DTIC] (D) and vindesine (V) with or without the addition of cisplatin (P). D was given intravenously (i.v.) at a dose of 250 mg/m2 days 1 5 every 4 weeks and V was given i.v. at a dose of 3.0 mg/m2 day 1 weekly. P was given i.v. at a dose of 100 mg/m2 day 1 every 4 weeks. There was no statistically significant difference in overall survival between the treatment arms (P = 0.22). Increased toxicity was observed in the treatment arm containing P of which leucopenia, alopecia and nausea/vomiting were the most pronounced. The median time to progression was significantly longer in patients treated with DVP (4.2 versus 2.2 months, P = 0.007). In conclusion, adding P to DV did not change overall survival but did significantly increase toxicity. PMID- 9849420 TI - Longitudinal evaluation of the complexed-to-total prostate specific antigen ratio in men with prostate disease. Effect of treatment. AB - The longitudinal changes in the complexed-to-total prostate specific antigen (PSA) ratio were evaluated in 90 men with benign prostatic hyperplasia (BPH) and 50 men with prostate cancer. The influence of treatment on this ratio was studied in 45 BPH patients and 50 patients with prostate cancer. Using a cut-off of 0.80 for the complexed-to-total PSA ratio, the large majority of prostate cancer patients had a ratio above the cut-off before treatment in serial determinations, whereas most BPH patients had a ratio consistently below that value. However, the few prostate cancer patients who had a ratio < or = 0.80 showed this low ratio in serial determinations, as did BPH patients who had a ratio > or = 0.80. During treatment, the ratio significantly decreased in 43 of the 50 patients with prostate cancer in parallel with the decrease in total PSA, and 34 of the 41 patients that had a pretreatment ratio > 0.80 showed a ratio < or = 0.80 during hormonal therapy. Our results show that neither the physiological changes in total and complexed PSA nor the treatment of BPH patients change the diagnostic efficacy of the complexed-to-total PSA ratio, whereas in prostate cancer patients under hormonal therapy, the ratio decreased in parallel with the decrease in total PSA. This suggests that, apart from improving the diagnostic efficacy of total PSA, the complexed-to-total PSA ratio could also be used to monitor BPH patients for newly developed tumours or to monitor therapy in patients with prostate cancer. PMID- 9849421 TI - Using reference data on quality of life--the importance of adjusting for age and gender, exemplified by the EORTC QLQ-C30 (+3). AB - Interpretation of health related quality of life (HRQOL) results in cancer patients is facilitated by knowledge of the levels of HRQOL in the general population. However, direct comparisons can be misleading unless age and gender are considered. We demonstrate the derivation of age- and gender-specific 'expected' values from population reference values by means of simple calculations. This survey included 3000 randomly selected Norwegians above 18 years of age who received the European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire (EORTC QLQ-C30 (+3) by mail. 1965 responses from 2,892 eligible persons (68%) were received. The population was divided into six disease groups based on self-reported health problems. The observed mean scale scores of the different groups deviated greatly from those obtained in the general population. The score for physical function, for example, was 72 for cancer patients and ranged from 73.3 to 82.5 in other disease groups, as opposed to 89.9 in the general population and 98.9 in those with no health problems. The range for one of the quality of life (QOL) scales was 57.7 to 84.7 compared with 73.7 in the general population. Expected mean scores for the patient groups were computed from the reference values, based on the concept of equivalence of age and gender. The differences between the observed mean scores and the reference values were strongly mediated by this method. The expected scores for physical function then ranged from 83.3 to 93.1 and from 70.3 to 75 for the QOL scale. The impact of age and gender on the reference data from the EORTC QLQ-C30 (+3) obtained in a general population shows that these variables must be considered when interpreting data on HRQOL for cancer patients. The demonstration of how to generate mean values which are adjusted according to the age and gender distribution of a population should increase the usefulness of this questionnaire among clinicians. PMID- 9849422 TI - 10 years' neuroblastoma screening in Europe: preliminary results of a clinical and biological review from the Study Group for Evaluation of Neuroblastoma Screening in Europe (SENSE). AB - Between January 1986 and May 1996, 870,313 children were tested in European neuroblastoma (NB) screening programmes. Among these children, 82 cases of NB (age range 4-24 months, median 11 months) were detected by screening. 83% of the patients had localised NB and 17% were diagnosed with generalised NB (stage 4, 10%; stage 4s, 7%). Unfavourable biological markers (MYCN amplification, loss of heterozygosity (LOH) 1p36, DNA di/tetraploidy) were observed in 14% of 76 biologically examined cases. The median follow-up time of all the patients was 21.5 months (range 1-101 months). To date, 69 patients are in complete remission (CR) and 2 patients have died due to therapy (stage 4, 1 patient; stage 3, 1 patient with unfavourable markers). Apart from screened patients, 16 other patients with NB were found who had previously had a normal screening test, i.e. 'false negative' patients (age range 10-41 months, median 31.5 months). The median interval between screening and diagnosis was 24.5 months (range 6-35 months). 11 of the 'false negative' patients suffered from generalised NB (stage 4) and 5 had localised NB at diagnosis. Unfavourable biological markers were observed in 7/12 patients. 5 patients have died, 2 achieved partial remission and 9 CR. 9 of the 11 patients with unfavourable biological markers diagnosed due to NB screening are currently in CR. It is very likely that, among the patients without unfavourable biological markers, we detected tumours which may have regressed spontaneously. These children may have undergone 'unnecessary,' but unavoidable, diagnostic procedures and therapy. To reduce the number of 'false negative' patients, a later screening could be helpful and should be evaluated. PMID- 9849423 TI - Treatment of neuroblastoma stage 4 with 131I-meta-iodo-benzylguanidine, high-dose chemotherapy and immunotherapy. A pilot study. AB - Disseminated neuroblastoma after infancy has a prognosis of approximately 10-20% with conventional therapy. We investigated the role of high-dose chemotherapy (HDCT) with peripheral blood stem cell (PBSC) rescue in combination with 131I metaiodobenzylguanidine ([131I-m]IBG). 11 children with neuroblastoma stage 4 were pretreated within the German Neuroblastoma Trial NB90 and included in a high dose concept for consolidation. Remission was documented by ultrasound, CT, NMR, or [123I-m]IBG scanning. HDCT was a combination of melphalan (180 mg/m2), carboplatin (1,500 mg/m2) and etoposide (40 mg/kg). All children were treated by [131I-m]IBG (0.58 GBq/kg) prior to high-dose treatment. All 11 children were additionally treated with antiGD2 murine- or chimeric-antibody (ch14.18). 4 children had no change to their remission status but three achieved a complete response (from a partial response to first line) and one a partial response (from no response to first line). The other 3 children progressed, 2 dying of their disease. Using Kaplan-Meier analysis, the probability of progression-free survival was 0.70 +/- 0.15 with a median observation time of 19 months. 9/11 children are alive, 8 without progression or relapse, whilst 2 have died of their disease. The combination of mIBG plus high-dose chemotherapy with PBSC support supplemented by immunotherapy with antiGD2 antibody appears to be a feasible and effective treatment regimen for disseminated neuroblastoma in this limited series. Larger numbers of patients should be treated to confirm these results. PMID- 9849424 TI - Alcohol consumption and risk of breast cancer: a multicentre Italian case-control study. AB - The relationship between alcohol consumption and breast cancer risk was investigated using data from a co-operative case-control study conducted in Italy between 1991 and 1994 on 2569 incident, histologically confirmed breast cancer cases and 2588 controls in hospital for acute, non-neoplastic, non-hormone related conditions. Overall, 915 (38%) cases and 1048 (43%) controls were abstainers. Compared with them, the odds ratio (OR), adjusted only for age, was 1.31 (95% confidence interval (CI) 1.13-1.53) for drinkers and became 1.39 (95% CI 1.(1)21-1.60) after correction for measurement error. The multivariate OR was 1.21 for drinkers of < or = 5.87 g/day and 1.23, 1.19, 1.21, 1.41 for drinkers of 5.88-13.40, 13.41-24.55, 24.56-27.60, > 27.60 g/day, respectively. The trend in risk was significant (chi 2 = 12.28, P < 0.0005). The association was apparently stronger in premenopausal women (OR = 1.80 for > 27.60 g/day). Considering the different types of alcoholic beverages (wine, beer, digestives, grappa and other spirits), a significant direct trend in breast cancer risk was seen for wine with an OR of 1.27 (95% CI 1.06-1.53) for the category > 26.34 g/day. The ORs were also above unity for beer, grappa, digestives and spirits drinkers. No appreciable interaction was observed between alcohol drinking and body mass index, smoking, or any other covariate considered. Thus, the present data, based on a validated alcohol consumption questionnaire and on a population characterised by a relatively high alcohol consumption in women, confirmed that alcohol drinking is moderately related to breast cancer risk. If causal, this association could explain 12% (95% CI, 5-19%) of breast cancers in Italy, thus representing one of the major avoidable risk factor for breast cancer. PMID- 9849425 TI - Screening for breast cancer in Ghent, Belgium: first results of a programme involving the existing health services. AB - In 1992, a population-based breast cancer screening programme was initiated in the municipality of Ghent: all women aged 40-69 years were personally invited to attend a local radiology unit for a mammogram, after being examined by a general practitioner or gynaecologist of their choice. The results of history taking, clinical breast examination, first and second reading, further investigation and primary treatment were registered. In total, 24.3% of the eligible population was screened in the period 1992-1994. The recall and biopsy rates were 2.9% and 1.4%, respectively. The cancer detection rate was 8.1 per 1000 women screened. Of all cancers detected, 88.0% (66/75) were invasive. Of these, 35.9% (23/64) measured 10 mm or less in diameter. The benign to malignant biopsy ratio was 0.7. Apart from the low participation, these results suggest that the programme is effective, compared with reference standards and the results of other studies. PMID- 9849426 TI - p53 expression in skin carcinogenesis and its relationship to cell proliferation and tumour growth. AB - The immunoreactivity of p53 protein was studied in relation to tumour development, histopathological characteristics, cell proliferation, and basement membrane organisation following the induction of skin carcinogenesis in tumour sensitive and -resistant mouse strains by ultraviolet (UV) irradiation or 7,12 dimethylbenz(a)anthracene (DMBA). In non-neoplastic skin exposed to UV irradiation or DMBA, p53 immunoreactivity was observed in nearly 50% of the basal layer cells. These cells were morphologically and histochemically indistinguishable from the p53-negative cells, occurring similarly in the tumour producing and the tumour-negative mouse strains and regardless of subsequent tumour formation. In induced epidermal hyperplasia and in benign tumours, p53 positive and proliferating cells constituted 40-50% of all cells in the basal layer, while superficial cells were p53 negative. In dysplastic epidermis, p53 positive cells and proliferating cells were seen in all cell layers. In the case of squamous cell carcinomas, p53-positive proliferating cells in differentiated neoplasms were localised close to the basement membrane and, more frequently, in border areas showing invasion and basement membrane destruction. In horn cysts, centrally located cells were non-proliferating and p53 negative. In moderately differentiated neoplasms, proliferating cells were located closer to the basement membrane, while p53-positive cells were distributed diffusely in the neoplasm. In poorly differentiated neoplasms, p53-positive cells were more common than proliferating cells and were arranged in a diffuse pattern. The results showed that the number and location of p53-positive cells depended upon histology, with a close relationship to tumour type and degree of malignancy, but not on the mode of induction, nor on the animal strain or the relationship to subsequent tumour formation. PMID- 9849427 TI - Regulation of c-fos transcription by chemopreventive isoflavonoids and lignans in MDA-MB-468 breast cancer cells. AB - Isoflavonoids and lignans are diet constituents with chemopreventive properties. We compared the ability of the isoflavonoids genistein and equol, the lignans enterodiol, enterolactone and nordihydroguaiaretic acid (NDGA) and the lignan metabolite methyl p-hydroxyphenyllactate to interfere with mitogenic and tumour promotional signal transduction pathways. Their effects on c-fos mRNA levels after induction by either epidermal growth factor (EGF) or the tumour promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) was measured in human breast cancer-derived MDA-MB-468 cells. Of the six agents, only genistein decreased EGF-induced, c-fos transcription (by 63% compared to control at 100 mumol/l). In contrast, both genistein and equol at 100 mumol/l decreased TPA induced c-fos levels, by 75 and 67%, respectively. NDGA and methyl p hydroxyphenyllactate did not inhibit TPA mediated c-fos transcription and enterolactone and enterodiol had only a weak inhibitory effect. NDGA at 0.1-10 mumol/l increased c-fos mRNA levels. None of the agents inhibited protein kinase C and only genistein inhibited EGF receptor-linked protein tyrosine kinase obtained from MDA-MB-468 cells, with an IC50 of 60 mumol/l. NDGA and genistein arrested cell colony formation potently, genistein was 15-fold more growth inhibitory than equol. The results suggest that both genistein and equol interfere similarly with TPA-induced signal transduction pathways. Inhibition by genistein of EGF-induced c-fos mRNA transcription is probably related to its interruption of EGF receptor-linked protein tyrosine kinase, whereas genistein induced growth arrest is not. If ability to antagonise phorbol ester effects is important for chemopreventive efficacy, equol and genistein might be equi efficacious chemopreventors, whereas enterolactone, enterodiol and NDGA should be much less potent. If phorbol ester antagonism together with antimitogenic activity determine optimal chemopreventive activity of this type of agent, genistein would be more potent than equol. PMID- 9849428 TI - Molecular characterisation of a panel of human ovarian carcinoma xenografts. AB - In a panel of 16 human ovarian tumours transplanted in nude mice, the expression of genes involved in cell cycle regulation and in response to drug treatment were characterised. In the 16 tumours analysed we could not detect overexpression of Erb-B2 oncogene while expression of MDR1 mRNA was not detected in 11/15 samples and was low in 4/15 tumours. Only three tumours had mutations in the p53 gene exons 5-8 and one of these mutations did not result in any amino acid alteration. The levels of mRNA for cyclins A, D1 and E were heterogeneous with some tumours expressing high levels and others not expressing them at all. The same was found for the cyclin dependent kinases (CDK) CDK2 and CDK4 and for CDK inhibitors p21/WAF1, p27/KIP1 and p16/CDKN2. Two genes belonging to the nucleotide excision repair, ERCC1 and ERCC3 were detectable in all the samples examined, as were the genes MGMT and MAG, also involved in DNA repair. The data indicate a heterogeneity in the expression of genes considered to be involved in the cellular responses to cytotoxic drug treatment and indicate the possibility of using these tumour models to test specifically molecules with a defined mechanism of action. PMID- 9849429 TI - Expression of two angiogenic factors, vascular endothelial growth factor and platelet-derived endothelial cell growth factor in human pancreatic cancer, and its relationship to angiogenesis. AB - Tumour angiogenesis, as assayed by microvessel density (MVD), and the expression of vascular endothelial growth factor (VEGF) and platelet-derived endothelial cell growth factor (PD-ECGF) have become established as important prognostic indicators for many tumour types. In this study, MVD and the expression of VEGF and PD-ECGF were examined by immunohistochemical staining of 50 pancreatic cancer tissues, and the relationships between either MVD or the expression of these two angiogenic factors and the clinicopathological features, including survival, were analysed. The expression of VEGF and PD-ECGF proteins were confirmed by Western blot analysis and VEGF mRNA isoforms were determined by reverse transcriptase polymerase chain reaction (RT-PCR) in five pancreatic cancer cell lines. Twenty eight (56%) of 50 pancreatic cancers were positive for VEGF protein in cancer cells, and 16 (32%) showed strong PD-ECGF staining in cancer and infiltrating cells. VEGF121 and VEGF165 were identified as the predominant species produced in pancreatic cancer cells. The overexpression of VEGF and PD-ECGF protein significantly correlated with high MVD (P = 0.002, 0.044, respectively). Advanced stage of disease was significantly more frequent in patients with high MVD (P = 0.025). No significant association was found between the expression of VEGF or PD ECGF and clinicopathological features, except for tumour histology. The expression of PD-ECGF correlated with poor survival (P = 0.011), but MVD and VEGF expression were not found to be useful for the prediction of overall survival. This study suggests that VEGF and PD-ECGF may play an important role in tumour angiogenesis, and that PD-ECGF expression seems to be useful for establishing prognoses for pancreatic cancer. PMID- 9849430 TI - Non-invasive in vivo mapping of tumour vascular and interstitial volume fractions. AB - Non-invasive measurement of haemodynamic parameters and imaging of neovasculature architecture is of importance in determining tumour prognosis, in directing tissue sampling and in assessing treatment efficacy. In the current research we investigated a dual tracer nuclear magnetic resonance (NMR) technique to map the tumour vascular (VVF) and interstitial volume fraction (IVF) non-invasively in vivo. We hypothesised that a NMR signal emanating after intravenous administrations of a vascular paramagnetic probe (MPEG-PL-GdDTPA) can be maximised so that additional signal after administration of a second interstitial probe (GdDTPA) would only reflect the IVF but not the VVF. The method and its assumptions were verified and experimental conditions optimised both in phantoms and in C6 glioma bearing rats. Data derived from in vivo studies show tumoral VVF and IVF values that are consistent with histology data and literature values; the relative ranking order of values was tumour > muscle > brain. Image maps showed intratumoral and intertumoral heterogeneity of both parameters at submillimetre pixel resolution. The method is applicable to a wide variety of tumour models and can theoretically be performed repeatedly to study tumour growth or involution during therapy. PMID- 9849431 TI - Cholecystokinin receptors in human pancreatic cancer cell lines. AB - Recent studies have suggested that cholecystokinin (CCK) receptors may play a role in the development and growth of pancreatic cancers. We detected the expression of mRNA encoding CCK-A and CCK-B receptors in eight human pancreatic tumour cell lines using reverse transcription-polymerase chain reaction (RT-PCR), but not by RNase protection assays. The K-ras gene, which can be activated by G coupled protein receptors such as CCK receptors, was mutated in codon 12 in five of the cell lines. In addition, Mia PaCa-2 pancreatic cancer cells did not respond to CCK or gastrin in cell proliferation or focal adhesion kinase (FAK) phosphorylation assays. In contrast, mouse NIH3T3 fibroblasts transfected with human CCK-B receptor (NIH3T3CCK-BR) showed increased proliferation and phosphorylation to the peptides. Also, radioligand binding studies indicated that Mia PaCa-2 cells had approximately 12.5-fold less CCK-B receptors than NIH3T3CCK BR. Our results suggest that in Mia PaCa-2 cells, CCK receptors may not play a crucial role in supporting cell growth. PMID- 9849432 TI - Urinary levels of cyclic guanosine monophosphate (cGMP) in patients with cancer of the uterine cervix: a valuable prognostic factor of clinical outcome? AB - Changes in urinary cyclic nucleotide levels have been reported in patients with various types of cancers. The present study was conducted to relate changes in urinary levels of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) to the clinical outcome of 11 patients treated for cancer of the uterine cervix. Urine was sampled for 24 h before and 3 months after primary treatment. The levels of cGMP increased in all the patients (n = 5) who relapsed within the observation period of 39 months. 4 of these patients showed an increased cGMP/cAMP ratio. In the patients without relapse (n = 6), the cGMP levels decreased, whereas the cGMP/cAMP ratios were unchanged. No marked changes in the levels of cAMP were observed for either of the groups. The measurement of urinary cGMP levels seems to be a valuable tool in the follow-up of patients with cancer of the uterine cervix. PMID- 9849433 TI - Phase II study of liarozole in advanced non-small cell lung cancer. AB - The aim of this phase II study was to investigate the efficacy and tolerability of liarozole, a novel benzimidazole derivative, in non-small cell lung cancer (NSCLC). Liarozole 300 mg twice daily orally was evaluated in 14 patients with stage IIIB and IV NSCLC. 8 patients had received prior treatment with chemotherapy and/or radiotherapy. WHO toxicity grading and response criteria were used. Liarozole was well tolerated. Grade 2 toxicities included alopecia (1 patient), dermatological toxicity (5 patients), dry mouth (2 patients) and nausea and vomiting (2 patients). Leukocytosis was seen in 5 patients, including 2 cases with an elevated white cell count pretreatment. Liarozole was discontinued in 1 patient who developed intolerable progressive pruritus associated with an erythematous rash. No objective tumour response was seen, all 14 patients developing progressive disease within 4 months of commencing treatment. Liarozole was well tolerated but was ineffective as single agent therapy in the management of NSCLC. The side-effect profile was compatible with inhibition of retinoic acid degradation. PMID- 9849434 TI - Comments on Management of cutaneous melanoma M0: state of the art and trends, Rossi et al., Eur J Cancer 1997, 33, 2302-2312. PMID- 9849435 TI - Extramedullary plasmacytoma of the larynx. Analysis of 5 cases. PMID- 9849436 TI - Small cell anaplastic carcinoma of the oesophagus following mantle field radiotherapy for mediastinal Hodgkin's lymphoma: first case report. PMID- 9849437 TI - Pre-operative CYFRA 21-1 levels in patients with lung cancer: correlation with mediastinal lymph node involvement. PMID- 9849438 TI - Second line treatment options in advanced colorectal cancer. PMID- 9849439 TI - Recombinant human erythropoietin (rhEPO) as an adjuvant for autologous blood transfusion. PMID- 9849440 TI - Should patients with advanced sarcomas be treated with chemotherapy? . PMID- 9849441 TI - The BRCA paradox in breast and ovarian cancer. PMID- 9849442 TI - Exemplifying guidelines for preparation of recombinant DNA products in phase I trials in cancer: preparation of a genetically engineered anti-CEA single chain Fv antibody. AB - Products of recombinant DNA technology have potential for the diagnosis or treatment of cancer. There is a need to investigate whether they function by the intended mechanism in small phase I clinical trials before their suitability for more extensive studies can be assessed. Quality and safety of these products should be assured prior to their use in humans in a way which is appropriate to the preliminary nature of the trials but not inhibitory to progress. The Cancer Research Campaign control recommendations for products derived from recombinant DNA technology (Begent RHJ and associates. Eur J Cancer 1993, 29A, 13, 1907-1910) provide guidelines for the production of new biotechnology products in academic research units within a relatively short time, while ensuring appropriate quality and safety. The practical application of the guidelines requires that solutions are found for the quality and safety issues during the production of recombinant products. We describe an approach to the relevant quality and safety issues during and after the production and purification of a genetically engineered anti carcinoembryonic antigen (CEA) single chain Fv (scFv) antibody for a phase I trial of radioimmunoguided surgery with the intention of providing a model for other products. PMID- 9849443 TI - The role of chemotherapy and radiation in the management of biliary cancer: a review of the literature. AB - Carcinoma of the biliary tract is a rare tumour. To date, there is no therapeutic measure with curative potential apart from surgical intervention. Thus, patients with advanced, i.e. unresectable or metastatic disease, face a dismal prognosis. They present a difficult problem to clinicians as to whether to choose a strictly supportive approach or to expose patients to the side-effects of a potentially ineffective treatment. The objective of this article is to review briefly the clinical trials available in the current literature utilising non-surgical oncological treatment (radiotherapy and chemotherapy) either in patients with advanced, i.e. locally inoperable or metastatic cancer of the biliary tract or as an adjunct to surgery. From 65 studies identified, there seems to be no standard therapy for advanced biliary cancer. Despite anecdotal reports of symptomatic palliation and survival advantages, most studies involved only a small number of patients and were performed in a phase II approach. In addition, the benefit of adjuvant treatment remains largely unproven. No clear trend in favour of radiation therapy could be seen when the studies included a control group. In addition, the only randomised chemotherapeutic series seemed to suggest a benefit of treatment in advanced disease, but due to the small number of patients included, definitive evidence from large, randomised series concerning the benefit of non-surgical oncological intervention as compared with supportive care is still lacking. Patients with advanced biliary tract cancer should be offered the opportunity to participate in clinical trials. PMID- 9849444 TI - Single agent infusional 5-fluorouracil is not effective second-line therapy after raltitrexed (Tomudex) in advanced colorectal cancer. AB - Raltitrexed (Tomudex) is currently licensed for first-line treatment of advanced colorectal cancer. We evaluated 101 patients treated with raltitrexed whose data were collected prospectively, in order to study the outcome of second-line treatments used after this drug. Of 98 evaluable patients, 50 received second line treatments, the commonest being 5-fluorouracil (5-FU)-based therapy (22 patients with 20 evaluable) and mitomycin-c (MMC) (22 patients with 18 evaluable). Only 1 response was seen in a patient treated with 5-FU and MMC and none following other treatments. This patient was not evaluable for outcome of raltitrexed treatment, having stopped after two courses. Patients who had responded to raltitrexed and later progressed off treatment were more likely to be offered second-line 5-FU, but despite the earlier sensitivity to thymidylate synthase inhibition, response rates were minimal. Underlying mechanisms for this lack of activity and proposals for future studies are discussed. PMID- 9849445 TI - Increased autologous blood donation in rectal cancer by recombinant human erythropoietin (rhEPO) AB - A randomised, placebo-controlled trial was conducted to study whether the subcutaneous administration of recombinant human erythropoietin (rhEPO) increases the donated red cell blood volume in patients with rectal cancer. Patients with resectable rectal cancer and a haemoglobin (Hb) level > or = 12.5/ > 12 g/dl (males/females) were scheduled to receive pre-operatively either erythropoietin (200 U/kg body weight daily) (n = 28) or placebo (n = 26) subcutaneously for 11 days. During this period autologous blood was collected. No serious adverse events were attributed to erythropoietin. 20 of 28 patients treated with rhEPO were able to donate > or = 3 units (71%) compared with 11 of 26 control patients (42%). The mean cumulative volume of red cells donated was 29% higher in the patients who received rhEPO (571 versus 444 ml, P = 0.02). The change in the mean reticulocyte value from baseline to the last pre-operative value was significantly higher in the rhEPO group (10.4 to 61.6/1000 versus 11.0 to 20.1/1000, P = 0.0001). The fall in the mean haematocrit from baseline to the last pre-operative value was significantly lower in the rhEPO group (41.4 to 37.6% versus 41.8 to 34.8%, P = 0.0004). rhEPO increases the ability of cancer patients to donate autologous blood during a short pre-operative period and enhances the restoration of haematological values after the donation period. PMID- 9849446 TI - Correlation between plasma transforming growth factor-beta 1 and second primary breast cancer in a chemoprevention trial. AB - The relationship between plasma transforming growth factor-beta 1 (TGF-beta 1) and second primary breast cancer was explored in a prevention trial of the synthetic retinoid fenretinide (N-(4-hydroxyphenyl)retinamide; 4-HPR). Plasma concentrations of TGF-beta 1 were measured by radioimmunoassay in plasma obtained at randomisation and after approximately 1 year of intervention in 28 women treated with 4-HPR and 27 untreated controls with stage I breast cancer. Baseline and 1 year growth factor concentrations were not significantly different between treated and control groups. After a median follow-up of 65 months, an increase in TGF-beta 1 over 1 year was the only significant, independent predictor of a shorter survival free from secondary primary breast cancer. Moreover, the change in TGF-beta 1 levels had a tendency to influence the treatment effect on second breast cancer incidence. Our data suggest that the role of plasma TGF-beta 1 as a surrogate endpoint of breast carcinogenesis should be assessed further. PMID- 9849447 TI - Entry into clinical trials in breast cancer: the importance of specialist teams. Scottish Breast Cancer Focus Group and Scottish Cancer Therapy Network. AB - The aim of this study was to identify the factors influencing entry of women with invasive breast cancer into clinical trials in Scotland. Women diagnosed during 1987 and 1993 were identified from cancer registry data records and their case notes reviewed. Entry into clinical trials was recorded, along with clinical and demographic data for 4688 patients. In 1987, the proportion of women entering clinical trials was 12.3% and, allowing for shorter follow-up, this appeared unchanged in 1993. Patients seen by surgeons with a high case load and those referred to an oncologist were approximately seven times and three times, respectively, more likely to enter a clinical trial (P < 0.0001). The area of Scotland (Health Board) where the woman was first treated also influenced study entry (P < 0.0001), whereas social deprivation had no effect (P = 0.93). Older women, especially those over 80 years of age, were less likely to enter studies (P = 0.05). Extending the management of patients by specialist multidisciplinary teams should increase recruitment into clinical trials and help to identify better treatments for women with breast cancer. PMID- 9849448 TI - A phase I trial of high-dose paclitaxel, cyclophosphamide and mitoxantrone with autologous blood stem cell support for the treatment of metastatic breast cancer. AB - The aim of this phase I study was to determine the dose limiting toxicity (DLT), maximum tolerated dose (MTD) and efficacy of a new combination of cyclophosphamide (6 g/m2), mitoxantrone (70 mg/m2), with dose escalation of paclitaxel (TaxolR) at a starting dose of 250 mg/m2 given intravenously over 3 h in a transplantation setting. Patients with metastatic breast cancer and chemosensitive disease were eligible. The autologous blood stem cell re-infusion and subsequent recovery occurred in an out-patient setting. 50 patients were enrolled, but 10 withdrew. 40 completed the entire protocol. At 400 mg/m2 paclitaxel administered over 3 h, 3 of 6 patients experienced serious adverse events: approximately 20-40 min after completion of infusion, diaphoresis, bradycardia mild hypotension and diarrhoea occurred; 2 patients lost consciousness for a few minutes. An extended infusion schedule delivering 400 mg/m2 paclitaxel over 6 h rather than 3 h was initiated at this level without patients experiencing this DLT. At the next dose of 450 mg/m2 paclitaxel over 6 h, the same DLT was seen as at 400 mg/m2 paclitaxel over 3 h and, therefore, MTD was reached. Time to recovery for the absolute neutrophil count > or = 0.5 x 10(9)/l was 10-19 days (median 12 days); and for platelets > or = 20 x 10(9)/l was 18-20 days (median 11.5 days). 21 patients developed neutropenic fever that required intravenous antibiotics and re-admission; the transfusion frequency for packed red blood cell was 0-5 units (median 2 units) and for platelets, 1-5 encounters (median 2). 13 complete responses, 1 patient with no evidence of disease and 19 partial remissions were documented. The dose of 400 mg/m2 at an infusion rate of 6 h will be used for the ongoing phase II study to evaluate efficacy and toxicity further. PMID- 9849449 TI - Altered expression of the growth and transformation suppressor PML gene in human hepatocellular carcinomas and in hepatitis tissues. AB - The promyelocytic leukaemia (PML) gene, which encodes a transformation and growth suppressor, was first identified at a chromosomal translocation break point in acute promyelocytic leukaemia. To elucidate if PML may be involved in hepatocellular carcinoma (HCC), the expression of PML was analysed using immunohistochemistry in human HCC and hepatitis tissues. Our studies demonstrated overexpression of PML protein in the PML-oncogenic domain (POD) structure in 50% of HCC (11/22). Enhanced expression and cytoplasmic localisation of PML was associated with cirrhosis. Increased expression of PML was also found in liver abscesses. However, in colon metastasis to the liver, the expression of PML was moderate to low, although strong expression was seen in the surrounding interstitial cells, macrophages and lymphocytes, an indication of the inflammation process associated with tumour growth. Most interestingly, strong expression of PML was found in neoplastic cells at the periphery of the tumours, but progressively decreased in cells at the centre of the tumours, which may be associated with an altered transform phenotype or apoptosis. The altered expression of PML indicates that this nuclear protein may play an important role in cellular response to stress and inflammation, as well as in compensatory cell growth. PMID- 9849450 TI - Clinical effects of human macrophage inflammatory protein-1 alpha MIP-1 alpha (LD78) administration to humans: a phase I study in cancer patients and normal healthy volunteers with the genetically engineered variant, BB-10010. AB - BB-10010 is a genetically engineered variant of human macrophage inflammatory protein-1 alpha (hMIP-1 alpha) with improved pharmaceutical formulation properties. Although initially described as a pro-inflammatory cytokine, it is now recognised that hMIP-1 alpha has additional effects on haemopoietic stem cell cycling and on human immunodeficiency virus uptake by macrophages. In view of the potential clinical utility of the molecule, we have embarked on a clinical trials programme to evaluate the safety, tolerability and haematological effects of BB 10010. We now report the results of two phase I clinical studies in which 49 subjects (9 patients with advanced breast carcinoma and 40 normal healthy volunteers) received escalating doses of BB-10010, from 0.1 to 300 micrograms/kg using the subcutaneous (s.c.) or intravenous route (i.v.) of administration. Treatment was associated with a dose-related increase in monocyte count which peaked at 200% of steady-state levels and was preceded by an acute, short-lived, monocytopenia, 50-100% of baseline. no measurable effects were noted on other leucocyte subsets or on circulating progenitor cell numbers. In all cases, BB 10010 was extremely well tolerated with no significant toxicity observed at any dose level and a maximum tolerated dose was not defined. Pharmacokinetic analysis revealed that serum concentrations of BB-10010 were detectable using doses of > or = 10 micrograms/kg i.v. or > or = 30 micrograms/kg s.c., and that a single s.c. injection resulted in sustained plasma levels over a 24 h period. These preliminary studies have confirmed the safety and tolerability of BB-10010 using a dose range up to 300 micrograms/kg. Further clinical studies are ongoing to determine the biological effects and to investigate the potential myeloprotective properties using a variable dose range and schedule of BB-10010 in combination with cytotoxic chemotherapy. PMID- 9849451 TI - Five days of oral topotecan (Hycamtin), a phase I and pharmacological study in adult patients with solid tumours. AB - Topotecan is a specific inhibitor to topoisomerase I. An oral formulation of topotecan is available with a bioavailability of 32-44% in humans. A phase I and pharmacological study of the oral formulation of topotecan administered daily for 5 days every 21 days was performed in adult patients with solid tumours to determine the maximum tolerated dose (MTD). Adult patients with a WHO performance status < or = 2 adequate haematological, hepatic and renal functions, with malignant solid tumours refractory to standard forms were entered into the study. Pharmacokinetics were performed on days 1 and 4 of the first course using a validated high performance liquid chromatographic assay. 29 patients entered the study, all patients were evaluable for toxicity and response. The doses studied in the 29 patients were 1.2, 1.8, 2.3, 2.7 mg/m2/day and a fixed dose of 4 mg/day without surface area adjustment. A total of 109 courses were given. Dose limiting toxicity (DLT) was reached at a dose of 2.7 mg/m2/day and consisted of CTC (NCI Common Toxicity Criteria) grade IV granulocytopenia. The regimen was well tolerated. Non-haematological toxicities were mild, including fatigue, anorexia, nausea, vomiting and diarrhoea. A significant correlation was observed between the percentage decrease in white blood cells versus the area under the curve (AUC(t)) of topotecan lactone (R = 0.76 P < 0.01) which was modelled by a sigmoidal Emax function. The correlation coefficient between the absolute topotecan dose administered and the AUC(t) was R = 0.52 (P = 0.04). Pharmacokinetics of the fixed dose of 4 mg/day were comparable to the 2.3 mg/m2/day dose. DLT in this phase I study of five daily doses of oral topotecan every 21 days was granulocytopenia. The recommended dose for phase II studies is 2.3 mg/m2/day or alternatively, a fixed dose of 4 mg/day. PMID- 9849453 TI - Retinoblastoma. PMID- 9849452 TI - Quality of life and survival in patients with advanced non-small cell lung cancer receiving supportive care plus chemotherapy with carboplatin and etoposide or supportive care only. A multicentre randomised phase III trial. Joint Lung Cancer Study Group. AB - The aim of the present trial was to evaluate the effects of chemotherapy on the quality of life and survival of patients with advanced non-small cell lung cancer (NSCLC) (stage IIIB or IV). In a controlled multicentre trial, patients were randomised to receive supportive care only or supportive care plus chemotherapy. Chemotherapy consisted of intravenous (i.v.) carboplatin 300 mg/m2 on day 1 and etoposide 120 mg/m2 orally on days 1-5 every 4 weeks for a maximum of eight courses. Quality of life was measured at randomisation and prior to each treatment course and at corresponding 4-week intervals in the control arm, using the EORTC QLQ-C30 + LC13 questionnaire. 48 patients were randomised (supportive care 26, chemotherapy 22), being eligible for comparative analyses. Another 102 patients, 97 of which received chemotherapy, were subsequently included in the study on an individual treatment preference basis. Data from these patients were used for confirmative purposes. Patients in the chemotherapy group reported better overall physical functioning and symptom control compared with the supportive care group. Group differences were smaller within the psychosocial domain, although trends were seen in favour of the chemotherapy group. No significant differences were seen in favour of the supportive care group, except for hair loss. Median survival times were 29 weeks in the chemotherapy group versus 11 weeks in the supportive care group, and 1-year survival rates were 28% versus 8%. Quality of life and survival outcomes were similar in the randomised and non-randomised patients receiving chemotherapy. No treatment-related deaths occurred. In conclusion, treatment with carboplatin and etoposide can improve both the quality of life and the survival of patients with advanced NSCLC. PMID- 9849454 TI - Treatment of non-metastatic rhabdomyosarcomas in childhood and adolescence. Results of the second study of the International Society of Paediatric Oncology: MMT84. AB - The second International Society of Paediatric Oncology (SIOP) study for rhabdomyosarcoma (MMT84) had several goals. The two principal aims were: (1) to improve the survival of children with rhabdomyosarcoma; and (2) to reduce the late effects from therapy by restricting the indications for surgery and/or radiotherapy after good response to initial chemotherapy. A further aim was to investigate the role of high-dose chemotherapy in young patients with parameningeal primary tumours. 186 previously untreated eligible patients entered the study. Patients with completely resected primary tumour received three courses of IVA (ifosfamide, vincristine and actinomycin D). Patients with incompletely resected tumour received six to 10 courses of IVA according to stage. Patients achieving complete remission with chemotherapy alone did not usually receive radiotherapy or undergo extensive surgery, but patients remaining in partial remission received local therapy with surgery and/or radiotherapy. Only patients over 5 years of age with parameningeal disease and patients over 12 years with tumours at any site were given systematic irradiation. Complete remission was achieved in 91% (170/186) of all patients. With a median follow-up of 8 years, the 5-year overall survival was 68% (+/- 3% standard error of the mean (SEM) and the 5-year event-free survival 53% (+/- 4% SEM). These results show an improvement over previous SIOP study (RMS75) in which survival was 52% and event-free survival was 47%. Among the 54 patients who exhibited isolated local relapse, 35% (19/54) survived in further remission longer than 2 years after retreatment, including local therapy (surgery +/- radiotherapy). Analysis of the overall burden of therapy received by all surviving children (including primary treatment and treatment for relapse if required) showed that 24% (28/116) were treated by limited surgery followed by three courses of IVA, 29% (34/116) were treated by chemotherapy alone (after initial biopsy) and 13% (15/116) received chemotherapy plus conservative local treatment (limited surgery or radiotherapy for residual disease). Only 34% (39/116) received intensive local therapy defined as radical wide field radiotherapy or radical surgery or both. Compared with the results obtained in the previous SIOP study, treatment in MMT84 was based on response to initial chemotherapy and, despite an overall reduction of the use of local therapy, significantly improved survival for patients with non-metastatic disease. This trial, also for the first time, provides evidence that retreatment after local relapse can achieve long-term second remissions. PMID- 9849456 TI - The influence of reproductive and hormonal factors on the risk of colon and rectal cancer in women. AB - A case-control study was conducted between 1992 and 1996 in six Italian areas. It included 537 women with colon cancer, 291 women with rectal cancer and 2081 control women in hospital for acute conditions, unrelated to hormonal or gynaecological diseases. A higher age at menopause was associated with increased colon cancer risk (odds ratio (OR) for > or = 53 years compared with < 50 years = 1.39, 95% confidence interval (CI) 1.04-1.87). Among parous women, a significant trend of decreasing colon cancer risk with increasing number of births was seen for colon (OR for > or = 4 births compared with 1 birth = 0.62, 95% CI 0.42 0.90), but not for rectal cancer. Nulliparous women, however, were at lower risk than women with a single birth, and age at first birth was directly associated with risk. While oral contraceptive use showed no significant influence, ever users of hormone replacement therapy had a reduced risk of rectal cancer (OR = 0.56, 95% CI 0.31-1.01). Thus, the association of colorectal cancer with reproductive and menstrual factors is neither strong nor consistent. PMID- 9849455 TI - An open-label, multicentre, randomised phase 2 study of recombinant human granulocyte colony-stimulating factor (filgrastim) as an adjunct to combination chemotherapy in paediatric patients with metastatic neuroblastoma. AB - Administration of combination chemotherapy to children with metastatic neuroblastoma induces profound myelosuppression resulting in chemotherapy treatment delays and febrile neutropenic episodes. The objective of this randomised multicentre study was to assess the incidence, duration and severity of neutropenia when filgrastim is added to induction chemotherapy administered to patients with metastatic neuroblastoma. In this study, 59 patients with metastatic neuroblastoma were randomised to receive chemotherapy (control group, n = 28) or chemotherapy plus filgrastim (filgrastim group, n = 31). Chemotherapy consisted of four cycles of cyclophosphamide, vincristine and doxorubicin (CADO) alternating at 21-day intervals with cisplatin and etoposide (CDDP-VP16). Filgrastim was administered subcutaneously at a dose of 5 micrograms/kg/day from day 7 for up to 14 days. The incidence of neutropenia (absolute neutrophil count [ANC] < 0.5 x 10(9)/l) in the filgrastim group was not reduced after the first CADO course. However, significant reductions were observed after courses 2, 3 and 4. The duration of neutropenia and of intravenous antibiotic use were significantly reduced in the filgrastim group over the whole study period (9 days versus 26 days, P < 0.001; 12 days versus 20 days, P = 0.04, respectively). However, the duration of hospitalisation and the incidence of febrile neutropenia were not significantly reduced. Compliance to treatment was good and the ability to administer chemotherapy without treatment delays was significantly better in the filgrastim group (P < 0.05). Event-free survival was greater in the filgrastim than in the control group (2.4 years versus 1.3 years; P = 0.072). Filgrastim is a beneficial adjunct to combination induction chemotherapy used in the treatment of metastatic neuroblastoma. PMID- 9849457 TI - Truncated TSG101 transcripts are present in peripheral blood from both familial breast cancer patients and controls. AB - TSG101 is a recently identified putative tumour suppressor gene which has been implicated in human breast cancer. To address whether germline disruption of TSG101 predisposes individuals to this disease, we analysed genomic DNA and mRNA isolated from peripheral blood from 20 familial breast cancer cases. No evidence of large intragenic insertions/deletions or point mutations in TSG101 was found by Southern blot analysis and sequence analysis of the entire coding region. However, in 11 of 20 samples, 'aberrant' transcripts were detected. Sequence analysis suggested that these variants were generated by the use of different cryptic splicing sites. Such alternative/aberrant splicing events were not restricted to cancer patients, but were also detected in peripheral blood of non cancer patients and in normal tissues. PMID- 9849458 TI - Intratumoral microvessel density and expression of ED-A/ED-B sequences of fibronectin in breast carcinoma. AB - The aim of this study was to examine the correlation between intratumoral microvessel density (iMVD) and the presence of cellular fibronectin isoforms, ED A and ED-B, in order to identify those tumours with a prominent angiogenic phenotype. 91 cases of invasive ductal breast carcinoma were evaluated for TNM, histological grading, percentage of Ki-67+ cells and receptor hormonal status. iMVD was determined as a single microvessel count in a 200 x microscope field from the region of the tumour that appeared to be most densely vascular. When the mean values of iMVD of the various groups were compared, no significant difference was noted (Mann-Whitney test). When tumours were classified as high or low iMVD, based on a cut-off value (99 vessels/0.74 mm2), cases with high iMVD were significantly more numerous in poorly differentiated G3 tumours (P = 0.01, Chi-square test), and in tumours with lymph node metastasis (N0 versus N1 + N2; P = 0.002). The possibility that high iMVD was the expression of prominent vascular neoformation was explored using ED-A and ED-B isoforms of fibronectin as markers of neoformed vessels. ED-A + and/or ED-B + blood vessels were < 10% of total vessels, were detected in approximately 50% of cases independently of iMVD values, and were not more numerous in tumour areas with hot spot vascularisation. Our findings indicate that iMVD and expression of ED-A/ED-B reflect different aspects of tumour-associated angiogenesis. PMID- 9849459 TI - Inhibition of growth of primary human tumour cell cultures by a 4 anilinoquinazoline inhibitor of the epidermal growth factor receptor family of tyrosine kinases. AB - The epidermal growth factor receptor (EGFR) is thought to mediate the action of the mitogens EGF and tumour growth factor-alpha (TGF-alpha) in a variety of cancers, including those of the lung, breast and ovary. A number of new selective inhibitors of EGFR tyrosine kinase have now been developed as potential new antitumour agents. We used a potent inhibitor of this tyrosine kinase, 6-amino-4 [(3-bromophenyl)amino]-7-(methylamino)quinazoline (SN 25531; PD 156273), to determine the responses of primary cultures derived from patients with cancer of the lung, ovary, breast, cervix and endometrium. Cells were cultured in 96-well plates and proliferation assessed by incorporation of 3H-thymidine. Measured growth inhibitory concentrations IC50 values) varied from 1 nM to 14 microM with a 1000-fold differential between sensitive and resistant cultures. Results were compared with rates of proliferation, estimated using a paclitaxel-based method. We also measured the IC50 values for the tyrosine kinase inhibitor using a number of established human cell lines, and compared them with EGFR content using fluorescent antibody staining and flow cytometry. The presence of EGFR was found to be necessary, but not sufficient, for in vitro response. Only a small number of cell lines (3 of 7 for lung, 1 of 7 for ovarian, 2 of 3 squamous cell and 0 of 12 for melanoma) were sensitive to the tyrosine kinase inhibitor. In contrast, 40 of the 50 primary cultures (including 14 of 15 lung cancer samples and 14 of 19 ovarian cancer samples) were sensitive. PMID- 9849460 TI - Effect of anthraquinone-laxatives on the proliferation and urokinase secretion of normal, premalignant and malignant colonic epithelial cells. AB - Even though 1,8-dihydroxyanthraquinone (DHA)-laxatives have been implicated in colon carcinogenesis, the available information is still inconclusive. The aim of this study was to demonstrate the effect of the DHA-laxatives, danthrone, rhein, aloe-emodin and sennidine, on colorectal tumour cells. In SW480 carcinoma cultures, dose-dependent induction of urokinase secretion into the medium was the predominant effect. Simultaneously, cell numbers were decreased by DHA-aglycones, but not by sennoside or the biphenylic laxative bisacodyl. DNA synthesis was not similarly reduced: 0.4-4 microM danthrone and sennidine even stimulated 5-bromo 2'-desoxyuridine (BrdU) uptake into DNA. When uptake was normalised to cell number, danthrone and sennidine doubled BrdU uptake/10(6) cells, 18 microM rhein and 0.7 microM aloe-emodin induced increases of 37 and 50%, respectively. This may at least partially be due to selective resistance of S-phase cells to DHA caused cell loss. In VACO235 adenoma cells, sennidine and aloe-emodin did not affect urokinase secretion, but stimulated growth. Both cell numbers and DNA synthesis were increased. In contrast to SW480 carcinoma cells, VACO235 cells were also sensitive to sennoside and bisacodyl. No effects of DHA were observed in normal colorectal epithelial cells. The biological effects were preceeded by specific phosphorylation of cellular proteins with molecular weights of 110, 78, 63, 57 kDa, indicating the specific induction of a cellular signalling cascade by the laxatives. PMID- 9849461 TI - No evidence for cancer-related CD44 splice variants in primary and metastatic colorectal cancer. AB - The expression of alternatively spliced CD44 adhesion molecules has been implicated in the pathogenesis and metastasis of colorectal cancer. Using a new set of primers for exon-specific reverse transcription-polymerase chain reaction (RT-PCR) we delineated the exact exon composition of CD44 mRNAs in normal colorectal mucosa, including isolated colonic crypts, in colorectal carcinomas and in their hepatic metastases. In addition, the surface expression of CD44 isoforms was analysed by immunohistochemistry. We identified by RT-PCR eight variant transcripts expressed in colorectal carcinomas and their metastases, but also constitutively in normal colorectal epithelia. In the normal colorectal epithelium, the surface expression of CD44 standard and variant molecules was restricted to proliferating cells at the bottom of the crypts. Despite expression of these transcripts in colorectal cancers and their metastases, monoclonal antibodies specific for standard or variant epitopes encoded by exons v5 and v6 stained only a few neoplastic lesions. These data point to a differentiation specific CD44 expression and splicing pattern in proliferating colorectal epithelia. However, they do not support a cancer- or metastasis-specific CD44 splicing pattern. Instead, cell surface availability of CD44 epitopes was reduced rather than increased in primary tumours and particularly in liver metastases. PMID- 9849462 TI - Flow cytometric analyses of the specific activation of peripheral blood mononuclear cells from healthy donors after in vitro stimulation with a fermented mistletoe extract and mistletoe lectins. AB - Immunostimulatory properties of mistletoe extracts derived from Viscum album L. (VAL) are well described, demonstrating activation especially of T, T-helper cells and monocytes/macrophages. In order to characterise in detail the communication between different cell populations, we studied mistletoe-induced expression of co-stimulatory signals and their ligands by flow cytometry. Peripheral blood mononuclear cells (PBMC) from 15 healthy controls were incubated for 7 days with a fermented VAL extract. VAL significantly upregulated the expression of the co-stimulatory molecule B7.1 (CD80) on monocytes/macrophages, but not B7.2 (CD86). No significant changes in the expression of either molecules on B cells could be found, suggesting that only monocytes/macrophages act as antigen presenting cells (APCs) in this in vitro system. Purified mistletoe lectins, components of most VAL extracts were also analysed, but did not induce similar responses of monocytes/macrophages. The receptor for B7 molecules, CD28, but not CTLA-4 (CD152), was also found to be significantly enhanced on CD4+ cells after VAL simulation. There was no evidence for activation of a B cell response via the CD40/CD40L pathway. Our data support the concept that stimulation by VAL extracts induces a specific T-helper cell reaction with monocytes/macrophages acting as APCs and purified lectins do not exert the same effects. PMID- 9849463 TI - A novel method to culture laryngeal human papillomavirus-positive epithelial cells produces papilloma-type cytology on collagen rafts. AB - A novel method to culture human papillomavirus (HPV) positive laryngeal epithelial cells is described. Biopsies of laryngeal papillomas and of HPV positive laryngeal mucosa were first cultured as a monolayer in which irradiated laryngeal fibroblasts originally derived from a papilloma (PPLF-XR) patient served as feeder cells. When these fourth or fifth passage epithelial cells were transferred to allow growth on an organotypic growth base (collagen raft containing unirradiated PPLF), they grew as a multilayer. This layer showed features typical of HPV infection with koilocytosis, parakeratosis, and isolated dyskeratotic cells. Based on in situ hybridisation, the original tumour sections and epithelial cells from each monolayer passage, as well as the collagen raft sections, contained HPV DNA. Our results show that HPV-infected epithelial cells can be maintained during passages in monolayer culture and that PPLF can support the growth of these cells well. The monolayer cell culture and the collagen raft, the latter providing differentiation-promoting effects, appears to facilitate maintenance of the infected cells and of the viral genome. PMID- 9849464 TI - Production of trypsins by human gastric cancer cells correlates with their malignant phenotype. AB - Proteolytic degradation of extracellular matrix is a critical step in tumour invasion and metastasis. To examine the role of trypsin in tumour dissemination, we cloned two variants (S4 and R3 cells) from STKM-1, a trypsinogen 1-producing diffuse gastric cancer cell line. Western blot analysis with antitrypsin antibody showed that 26 and 24 kDa proteins were highly detected in S4 conditioned medium (CM) in comparison to R3 CM. In addition to the 26 and 24 kDa proteins, 25 and 23 kDa bands, which correspond to enterokinase-activated trypsin, were found only in S4 CM. When the CMs of the two clones were treated with enterokinase, the 25 and 23 kDa trypsin activities in S4 CM were effectively increased as compared with R3 CM. When the two clones were inoculated intraperitoneally (i.p.) into nude mice, S4 cells strongly invaded the liver, pancreas and peritoneum and killed the hosts more rapidly than R3 cells: the 50% survival time was 50 days for S4 and 82 days for R3 cells. These results suggest that trypsin production is associated with the invasive growth of STKM-1 gastric cancer cells. PMID- 9849465 TI - Phase II study of oxaliplatin in poor-prognosis non-small cell lung cancer (NSCLC). ATTIT. Association pour le Traitement des Tumeurs Intra Thoraciques. AB - The aim of this phase II study was to determine the antitumour activity and safety of trans-1-diaminocyclohexane-platinum (oxaliplatin) in previously untreated advanced non-small cell lung cancer (NSCLC) patients. 33 patients with unresectable and measurable NSCLC were entered into this phase II study between January 1992 and January 1994. Patients had either locoregional disease with performance status 2 (19 patients) or a stage IV disease (14 patients). Oxaliplatin (130 mg/m2) was given on an out-patient basis (2-h infusion, every 21 days) without hydration. Response was assessed after every two courses. One hundred courses were administered, with a mean of three courses per patient (range 1-12). All patients were evaluable for response; 1 had a complete response, and 4 a partial response (overall response rate 15%, 95% confidence interval 5.1-31.9%). The median response duration was 5.9 months. All cycles (n = 100) were evaluable for toxicity assessment. Transient reversible, cold-related finger dysesthesias occurred in 29 patients, but were mild, and disappeared in most cases within a few days. We observed brief episodes of pharyngolaryngeal discomfort (8 patients, 11 episodes) accompanied in 4 cases (3 patients), by transient episodes of inspiratory stridor, leading 2 patients to treatment withdrawal. We conclude that oxaliplatin has activity in poor-prognosis NSCLC and that this treatment is feasible in out-patients; the absence of renal and haematological toxicity makes this drug a good candidate for further evaluation in NSCLC. PMID- 9849466 TI - Treatment of advanced gastric cancer with oral etoposide, leucovorin and tegafur: experience with an oral modification of the etoposide, leucovorin and 5 fluorouracil (ELF) regimen. AB - Recent data have suggested enhanced therapeutic activity with prolonged administration of both etoposide as well as fluoropyrimidines in the treatment of gastrointestinal malignancies. Based on this rationale, we investigated the clinical effectiveness and tolerance of an oral modification of the widely applied etoposide, leucovorin and 5-fluorouracil (ELF) regimen in patients with advanced gastric cancer. 32 patients with advanced gastric cancer were treated with oral etoposide (100 mg), leucovorin (3 x 100 mg), and tegafur (3 x 200 mg) over 14-21 days for a maximum of six cycles. Objective response was seen in only 5 patients (16%), stable disease was documented in 7 (22%), while the remaining patients progressed during therapy. The median time to progression was 2.8 months (range 0.7-12 months) and median overall survival was 6 months (range 1-18+ months). Due to grade 3 nausea/emesis, 8 patients discontinued treatment prematurely, while 12 patients experienced anorexia and progressive weight loss. Haematological toxicity was modest, with 4 patients developing asymptomatic grade 3-4 granulocytopenia. We conclude that this oral combination regimen cannot be recommended for the treatment of advanced gastric cancer. PMID- 9849467 TI - A phase II study of DaunoXome in advanced urothelial transitional cell carcinoma. AB - Liposomal encapsulation of anthracyclines is claimed to reduce toxicity and to improve pharmacokinetics. Therefore, 15 patients with locally advanced or metastatic transitional cell cancer (TCC) of the urinary tract were entered into a phase II study assessing the response rate (WHO criteria) and toxicity of DaunoXome 100 mg/m2 given as a 1 h infusion every third week. During treatment, 6 patients remained stable and 8 had progressive disease. 1 patient died of pulmonary embolism after the first cycle and was not evaluable for response. No patient developed grade 4 myelotoxicity. Grade 3 leucopenia was seen in 5 patients and grade 1 thrombocytopenia in 1 patient, with no treatment-related changes of biochemical liver and kidney function tests. 4 patients complained of angina pectoris-like chest pain during the initial phase of the first or second infusion. The event was associated with a decrease in systolic blood pressure by 20-30 mm in 1 patient leading to permanent treatment discontinuation. In the other 3 and all subsequent patients, intramuscular application of 100 mg hydrocortisone 1 h prior to DaunoXome infusion prevented similar hypotensive reactions. In this study, intravenous (i.v.) DaunoXome 100 mg/m2 every third week showed no anticancer activity in advanced TCC. PMID- 9849468 TI - A case of ototoxicity in a patient with metastatic carcinoma of the breast treated with paclitaxel and vinorelbine. PMID- 9849469 TI - Comments on inhibition of growth of androgen-independent DU-145 prostate cancer in vivo by luteinising hormone-releasing hormone antagonist Cetrorelix and bombesin antagonists RC-3940-II and RC-3950-II, Jungwirth et al., Eur J Cancer 1997, 33(7), 1141-1148. PMID- 9849470 TI - Clinical and biological implications of the Milan breast conservation trials. PMID- 9849472 TI - Should maintenance chemotherapy be used to treat small cell lung cancer? PMID- 9849471 TI - Actinomycin D, hepatic toxicity and Wilms' tumour--a mystery explained? PMID- 9849473 TI - Ten year results of a randomised trial comparing two conservative treatment strategies for small size breast cancer. AB - We report the 10-year results of a randomised clinical trial in which two different breast conservation treatment strategies were compared in women with small, non-metastatic primary breast cancer: quadrantectomy, axillary dissection and radiotherapy (QUART) versus tumorectomy and axillary dissection followed by external radiotherapy and a boost with 192Ir implantation (TART). No second surgery was given to women with affected surgical margins. Axillary node positive women received adjuvant medical therapy. From 1985-1987, this trial accrued 705 patients, 360 in the QUART and 345 in the TART arm. Crude cumulative incidence curves for intrabreast tumour recurrence (IBTR) and metastases as first events and mortality curves in each of the two treatment arms were computed. A crude cumulative incidence curve of IBTR as a second event (in women who had already had a local recurrence) was also computed. The two groups were compared in terms of hazard for IBTR, metastases or death occurrence by using Cox regression models, both with and without adjustment for patient age, tumour size, number of metastatic axillary nodes and histology. Possible interactions between the aforementioned prognostic factors and the type of surgery were also investigated. The two groups were well matched for baseline patient and tumour characteristics, the only exception being resection margins, which were more often positive in the TART group. At the Cox model, a significant difference between groups was detected for IBTR (P < 0.0001), but not for distant metastases and overall survival. In particular, 5- and 10-year estimates of crude cumulative incidence of IBTR were 4.7 and 7.4% in the QUART group and 11.6 and 18.6% in the TART group. The difference was not substantially affected by patient or disease characteristics. Likewise, the status of resection margins in women who underwent TART treatment did not significantly influence the risk of occurrence of IBTRs. Finally, the rate of second IBTR occurrence was relatively high, when compared with the rate of IBTR occurrence as first event. In summary, the results of this trial show that a better local control of the disease can be obtained with the more extensive surgical resection, i.e. QUART. PMID- 9849474 TI - A woman's build and the risk of breast cancer. AB - A woman's build and the risk of breast cancer seem to be related. While relative overweight, as described by the body mass index, seems to be associated with increased breast cancer risk in postmenopausal women, overweight in premenopausal women seems slightly protective. Papers from a MEDLINE search are reviewed regarding the association between build and the development of breast cancer. Different aspects of build, such as height, weight, body mass index and body shape, are discussed. The more prominent associations found through this search are a positive association between height and breast cancer risk both in pre- and postmenopausal women. Regarding body mass index, the association is negative in premenopausal women and positive in postmenopausal women. Body shape described as masculine versus feminine seems to have no impact on breast cancer risk in premenopausal women, but seems to be positively associated with breast cancer in postmenopausal women. Possible biological mechanisms responsible for the associations with breast cancer risk are discussed, including endogenous oestrogens, androgens and glucose metabolic substances. Avoiding or reducing postmenopausal overweight may modify breast cancer risk indicators in a more favourable direction. PMID- 9849475 TI - Ten-year results of a randomised trial comparing cisplatin with cisplatin and cyclophosphamide in advanced, suboptimally debulked ovarian cancer. AB - 176 eligible patients with advanced suboptimally operated ovarian carcinoma were randomly allocated to receive either cisplatin 75 mg/m2 or cisplatin 50 mg/m2 and cyclophosphamide 500 mg/m2 (CP) every 28 days for six courses. The overall clinical response rates (complete response plus partial response) were 52 and 63% for CP and cisplatin, respectively (non-significant). Including results obtained by second-look laparotomy, we did not observe a statistically significant difference in response rates in the two treatment groups. Median progression-free survival was 10 and 11.9 months for CP and cisplatin, respectively (non significant). No significant difference was observed in overall survival, with a median of 19.4 and 21.5 months for CP and cisplatin, respectively. Thirty-seven platinum-resistant and 27 platinum-sensitive tumours were treated with carboplatin or cisplatin as second-line therapy. Response rates to platinum second-line therapy were 6 and 50% for resistant and sensitive tumours, respectively (P < 0.001). This difference in response rate was also confirmed by survival analysis. Patients with platinum-sensitive tumours survived longer when they were treated with platinum-containing chemotherapy (P = 0.005). Median survival was 22.8 and 8.5 months after initiation of second-line treatment for the platinum-containing and platinum-free regimens, respectively. In summary, we observed in suboptimally operated ovarian carcinoma patients similar response rates, progression-free interval, and overall survival for equitoxic cisplatin and CP. However, the doses of cisplatin and cyclophosphamide chosen were substantially lower than current standard doses of CP. Our study demonstrates, therefore, that a suboptimal dose of CP is as effective as optimal dose monotherapy cisplatin. Patients with recurrences considered as platinum-sensitive had a significantly higher response rate and improved survival when retreated with platinum-containing therapy. PMID- 9849476 TI - Quality of life assessment in daily clinical oncology practice: a feasibility study. AB - Quality of life (QL) assessments are increasingly being included in clinical trials, but their use in clinical practice is still uncommon. The objectives of this study were to investigate the feasibility of introducing individual QL assessments into the daily routine of an out-patient oncology clinic, and the potential impact of such assessments on doctor-patient communication. The study sample included six physicians and 18 of their patients from the out-patient clinic of the Netherlands Cancer Institute/Antoni van Leeuwenhoek Hospital in Amsterdam, The Netherlands. For each patient, three follow-up consultations were observed. The first visit was employed for the purpose of a baseline measurement. At the two subsequent visits, the patients were asked to complete the EORTC QLQ C30, a standardised cancer-specific QL questionnaire. The patients' responses were computer-scored and transformed into a graphic summary. The summary included current scores as well as those elicited at the previous visit. Both the physicians and the patients received a copy of the summary just prior to the medical consultation. Completing, scoring and printing the QL data could be done during waiting room time. The availability of the summary did not lengthen the average consultation time. A small increase was noted in the average number of QL issues discussed per consultation. However, the most notable trend was the increased responsibility taken by the physicians in raising specific QL issues for discussion. When the QL summary was available, the physicians raised three times as many topics than was the case prior to its use (P < 0.05). All six physicians and the majority of patients believed that the QL summary facilitated communication, and expressed interest in continued use of the procedure. The introduction of individual QL assessments in routine out-patient oncology practice is feasible and appears to stimulate physicians to inquire into specific aspects of the health and well-being of their patients. However, given the methodological limitations of this pilot study, the results should be interpreted with caution. PMID- 9849477 TI - Radiosurgery without whole brain radiotherapy in melanoma brain metastases. Club de Cancerologie Cutanee. AB - To evaluate the effectiveness of radiosurgery without whole brain radiotherapy in the palliative treatment of melanoma brain metastases, we retrospectively assessed the results in 35 patients: 4 with a solitary brain metastasis, 13 with a single brain metastasis and metastases elsewhere and 18 with multiple brain metastases. The local control rate was 98.2% (55/56 metastases) at 3 months. Median survival was 22 months in patients with a solitary brain metastasis, 7.5 months in patients with a single brain metastasis and metastases elsewhere, and 4 months in patients with multiple brain metastases. Complications were unusual and surgery was required in 2 of 35 patients. These results show for the first time that melanoma patients with a unique brain metastasis with or without metastases elsewhere clearly benefit from tumour control easily obtained by radiosurgery. Although the comparison of radiosurgery with surgery and/or whole brain radiotherapy cannot be adequately addressed, radiosurgery alone seems to provide similar results with lower morbidity and impact on quality of life. PMID- 9849478 TI - Urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI-1) in tissue and serum of head and neck squamous cell carcinoma patients. AB - The aim of this study was to determine urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI-1) concentrations in tumour and adjacent normal tissue samples from 58 patients, and in serum samples from 40 of 58 patients with squamous cell carcinoma of the head and neck obtained at diagnosis and after completion of therapy. uPA and PAI-1 serum concentrations were also measured in 28 healthy volunteers who served as controls. Measurements were made using enzyme-linked immunosorbent assay (ELISA) techniques. For both uPA and PAI-1, significantly elevated concentrations were measured in tumour tissue as compared with normal tissue (uPA: 8.89 versus 0.41 ng/mg total protein (mgp), P < 0.0001; PAI-1: 23.9 versus 1.47 ng/mgp, P < 0.0001). A statistically significant difference in uPA concentrations was found between normal laryngeal and nonlaryngeal tissue (0.52 versus 0.3 ng/mgp, P = 0.008), and in PAI-1 concentrations between T1 + 2 and T3 + 4 stage of disease (17.32 versus 35.63 ng/mgp, P = 0.04). The uPA concentrations positively correlated with those of PAI 1 measured in both tumour (Rs = 0.62, P < 0.0001) and normal tissue (Rs = 0.30, P = 0.02). In serum samples, lower concentrations of PAI-1 were measured in the control group than in patients with cancer (412.0 versus 680.5 ng/ml serum (mls), P = 0.0006). The time of collection of the serum sample did not influence uPA and PAI-1 concentrations, and no association was observed between their concentrations and any clinical and histopathological prognostic factors tested. Our results indicate that both uPA and PAI-1 may play a specific role in the process of invasion and metastasis, and might also be of prognostic value in squamous cell carcinoma of the head and neck. PMID- 9849479 TI - Clinical evaluation of autoantibodies to p53 protein in patients with chronic liver disease and hepatocellular carcinoma. AB - In hepatocellular carcinoma (HCC) of patients from the Western hemisphere, mutations in the p53 tumour suppressor gene are present in up to 37% of cases. Conformational change and cellular accumulation can initiate an immune response with generation of circulating autoantibodies to p53 protein. In the present study, we investigated 711 consecutive patients with chronic liver disease to evaluate the sensitivity and specificity of autoantibodies to p53 protein as a serological marker for HCC. Detection of p53 autoantibodies was performed using an enzyme-linked immunosorbent assay with immobilised recombinant p53 protein. Liver cirrhosis was present in 259 patients (36.4%) and a HCC was diagnosed in 75 patients (10.6%). Autoantibodies to p53 protein were detectable in 15 of 377 patients with chronic liver disease (4.0%) and in 10 of 259 patients presenting with liver cirrhosis (3.9%). All 25 p53 autoantibody-positive/HCC-negative patients were carefully investigated and no underlying malignancy was clinically detected, suggesting that elevated p53 antibody levels may not exclusively be detectable in patients with malignant disease. In patients with clinically manifest HCC, p53 autoantibodies were detected in 17 of 75 cases, thus resulting in a sensitivity of 22.7% and a specificity of 96.1%. In contrast, assessment of serum alpha-fetoprotein (AFP) resulted in a sensitivity and specificity of 69.3 and 91.8% (AFP > 20 ng/ml) and 53.3 and 99.1% (AFP > 100 ng/ml) for the detection of HCC, respectively. The data of the present study reveal that the presence of p53 autoantibodies in patients with chronic liver disease is not completely specific for HCC. Moreover, we obtained no direct evidence that p53 autoantibody formation precedes the clinical diagnosis of HCC. However, serological screening for HCC might be improved by combining AFP and p53 autoantibody assays. PMID- 9849480 TI - Chemotherapy for gestational trophoblastic tumours hastens menopause by 3 years. AB - Chemotherapy may induce acute ovarian failure, but in women who retain gonadal function throughout chemotherapy, the late effects upon ovarian function are unknown. A retrospective controlled survey was performed to ascertain whether chemotherapy for gestational trophoblastic tumours (GTT) results in premature menopause. Questionnaires were sent to 1,489 women diagnosed between 1971 and 1990 with GTT, including 1089 who had received chemotherapy and 400 who had not received chemotherapy (controls). Responses were obtained from 972 chemotherapy treated patients and 327 controls. 124 women were not evaluable for menopause date as they had undergone hysterectomy as part of the treatment for GTT or had developed permanent amenorrhoea during chemotherapy. Overall, 172 women reported that they were postmenopausal, including 157 women who had received chemotherapy. The median age at menopause for the evaluable population was 50 years (range 25 56 years). The age at menopause was significantly earlier in the treated arm (median 50, range 25-56 years) than in the controls (median 53, range 40-57 years) (logrank test chi 2 = 12.6, P = 0.0004). Menopause occurred significantly earlier in women treated with combination chemotherapy (median 49, range 25-56 years) compared with single agent methotrexate (median 51, range 25-56 years) (logrank test chi 2 = 8.3, P = 0.004). However, the age at completion of chemotherapy in the treated arm did not influence the age of menopause (proportional Hazards chi 2 = 1.99, P = 0.16). Chemotherapy for GTT induces menopause 3 years earlier than it occurs in women with GTT who do not receive chemotherapy. Although the difference is statistically significant, the magnitude is modest and most women can be reassured that neither fertility nor postmenopausal osteoporosis will be greatly affected. PMID- 9849481 TI - A phase II study of gemcitabine in patients with transitional cell carcinoma of the urinary tract previously treated with platinum. Italian Co-operative Group on Bladder Cancer. AB - The aim of this study was to evaluate the efficacy and safety of gemcitabine, a pyrimidine antimetabolite, in the treatment of advanced transitional cell carcinoma of the urinary tract. 35 patients with unresectable or metastatic transitional cell carcinoma of the urinary tract previously treated with a platinum-based regimen were studied. Gemcitabine was administered at a dosage of 1200 mg/m2 as a 30-min intravenous infusion on days 1, 8 and 15, repeated every 28 days. 31 patients were evaluable for efficacy. 4 patients achieved a complete response (12.9%), 3 a partial response (9.6%) and 13 (42%) were stable for at least 4 weeks (overall response 22.5%; 95% confidence interval 8-37%). The median response duration was 11.8 months (range 3.6-17.7 + months) and median survival for all patients entered was 5 months (range 2-21 + months). 2 patients with complete response are still alive with no evidence of disease after 14 and 21 months. Gemcitabine also provided subjective symptomatic relief from pain, cystitis, dysuria, haematuria and peripheral oedema. Patients experienced little WHO grade 3-4 toxicity, with anaemia in 8 patients (23%), thrombocytopenia in 5 (14.2%), leucopenia in 4 (11.4%) and neutropenia in 7 (20%). WHO grade 3-4 hepatic toxicity occurred in 4 patients (11.4%) and transient elevations of transaminase was noted in 3 (8.6%). No patient had WHO grade 3-4 elevation of serum creatinine level. There was no WHO grade 4 symptomatic toxicity and no alopecia was noted. Transient influenza symptoms with gemcitabine occurred in 18 patients (51.4%) with 13 patients (37.1%) experiencing fever (2.9% WHO grade 3). In conclusion, gemcitabine is an new active agent for the treatment of transitional cell carcinoma of the urinary bladder with a mild toxicity profile; it warrants further investigation in combination with cisplatin in chemotherapy naive patients. PMID- 9849482 TI - Electrochemotherapy with cisplatin: potentiation of local cisplatin antitumour effectiveness by application of electric pulses in cancer patients. AB - This study was aimed at assessing the response to electrochemotherapy with cisplatin of cutaneous tumour nodules in patients with malignant melanoma, squamous cell carcinoma and basal cell carcinoma. In 4 patients, 30 tumour nodules of different sizes were treated; five without treatment, one with electric pulses, five with cisplatin injected intratumorally and 19 with electrochemotherapy, i.e. intratumoral administration of cisplatin followed by delivery of electric pulses to the tumour nodule. After 4 weeks, a complete response (CR) in all 19 electrochemotherapy treated nodules was obtained. All electrochemotherapy treated nodules remained in CR (range 7-11 months), regardless of histological type, except for the metastasis of a squamous cell carcinoma that progressed after 9 months. CR was also obtained in two of five tumour nodules treated with cisplatin intratumorally, but the other three nodules progressed within 3-7 months. Exposure of the tumour nodule to electric pulses without cisplatin treatment had no effect on tumour growth. Electrochemotherapy was well tolerated by all patients and a good cosmetic effect was obtained, with only minimal scarring and a slight depigmentation of the skin. Electrochemotherapy with cisplatin has proved to be effective in patients with cutaneous tumour nodules. Furthermore, electrochemotherapy is easy to perform and can be carried out on an out-patient basis. PMID- 9849483 TI - Veno-occlusive disease of the liver in right-sided Wilms' tumours. AB - Veno-occlusive disease of the liver (VOD) is an important complication in children with Wilms' tumour. Although in most patients this complication resolves uneventfully, fatal cases have been reported. Several observations strongly suggest that actinomycin-D is the likeliest cause of VOD in Wilms' tumour, but VOD seems to be rather uncommon in other malignancies treated with chemotherapy including actinomycin-D. The present case of VOD and the review of the literature stress the pathogenetic and clinical implications of VOD in the presence of a Wilms' tumour treated with actinomycin-D, originating in the right kidney. Greater awareness of this 'predisposing factor' may alert paediatricians to the presence of minimal signs of the syndrome. PMID- 9849484 TI - Treatment of intermediate risk rhabdomyosarcoma and undifferentiated sarcoma with alternating cycles of vincristine/doxorubicin/cyclophosphamide and etoposide/ifosfamide. AB - Over 50% of patients with newly diagnosed rhabdomyosarcoma (RMS) are in the 'intermediate risk' group with a 3-year progression-free survival of approximately 65%. This group consists of stage 1, group III, non-orbit tumours; stage 2, group II and III; and all stage 3 patients utilising the Intergroup Rhabdomyosarcoma Study (IRS) staging system. The role of doxorubicin in the treatment of RMS has been controversial. Ifosfamide, both alone and in combination with etoposide, has significant activity in patients with RMS. The aim of this pilot study was to examine the efficacy and toxicity of a chemotherapy regimen of alternating cycles of vincristine/doxorubicin/cyclophosphamide and etoposide/ifosfamide for intermediate risk RMS. 30 patients with intermediate risk RMS or undifferentiated sarcoma (US) were treated with alternating cycles of vincristine/doxorubicin/cyclophosphamide (VDC) and etoposide/ifosfamide (EI) at planned intervals of 3 weeks. Local treatment of the tumour in most cases was performed after four cycles of chemotherapy, followed by an additional 10 cycles of chemotherapy. At a median follow-up of 37.5 months, the Kaplan-Meier estimate of 3-year event-free survival was 85% (95% confidence interval 72-99%). The overall survival at 3 years was 91% (95% confidence interval 80-100%). No patient died from toxicity. The most common toxicity was febrile neutropenia in 35% of VDC and 26% of EI cycles. No nephrotoxicity or cardiac toxicity was seen. No patient progressed prior to week 12 local therapy. Alternating cycles of VDC and EI are an effective treatment for patients with intermediate risk RMS and US. Toxicity is tolerable. Delaying local treatment until week 12 does not compromise outcome. PMID- 9849485 TI - Risk factors for HPV detection in archival Pap smears. A population-based study from Greenland and Denmark. AB - The most important risk factor for cervical cancer is genital infection with certain types of human papillomavirus (HPV). The presence of HPV was studied in archival smears from a random sample of women living in Greenland (GW) and Denmark (DW) having, respectively, a high risk and an intermediate risk for cervical cancer. Risk factors were also examined of the original 126 Danish and 129 Greenlandic archived smears collected during October and November 1988. 125 were located from each country including all abnormal smears. HPV DNA was isolated from the smears and detected by means of a consensus polymerase chain reaction (PCR) detecting a broad spectrum of genital HPV types. HPV was detected in all the abnormal smears and in 22 and 33% respectively of the cytological normal smears from DW and GW. Risk of HPV was significantly higher in smears from women who started sexual life relatively recently (respectively, < or = 4 and < or = 6 years ago in DW and GW) compared with > or = 10 years ago (adjusted prevalence-OR: 9.3; 95% CI: 2.2-39.2 in DW and 5.9; 95% CI: 1.4-25.3 in GW). Among other important risk factors were age in both areas, lifetime number of sex partners and current smoking in DW and ever and gonorrhoea in GW. This study confirms the usefulness of the method as all abnormal smears were positive and, furthermore, the predictors for HPV presence in the normal smears corroborate with those found in recent studies of HPV in fresh cervical swabs. Thus, this method can be useful for large-scale epidemiological studies of HPV DNA in already sampled material. PMID- 9849486 TI - Trends in treatment and long-term survival of thyroid cancer in southeastern Netherlands, 1960-1992. AB - Thyroid cancer (TC), comprising less than 1% of all cancers in the Netherlands, has a good prognosis in general. Controversy still remains on the extent of surgical treatment and the indication for additional Iodine-131 (131I) therapy in the management of differentiated TC. The aim of this study was to describe (changes in) the treatment of TC and to determine independent prognostic factors for crude and relative survival of differentiated TC diagnosed in general hospitals. This population-based, retrospective study was based on data from the Eindhoven Cancer Registry, Comprehensive Cancer Centre South (I.K.Z.), Eindhoven, the Netherlands. Data were collected on all 343 TC patients diagnosed from 1 January 1960 to 31 December 1992. All available information on treatment (initial and additional) and survival (on 1 April 1994) were recorded. Initial surgical treatment was defined as limited or extended. Multivariate analysis of crude and relative survival to determine prognostic factors for differentiated TC was performed. Mean follow-up was 7.6 years. The proportion of patients with differentiated TC increased from 60% in 1960-1972 to 84% in 1985-1992. TC patients were treated in all hospitals in the region, approximately 2-4/year. Ninety per cent of all TC patients initially underwent surgical treatment; the extended procedures increasing from 27% in 1960-1974 to 61% in 1985-1992. 131I was also administered increasingly (from 18-44%) to patients with differentiated TC. The relative 5, 10 and 20 year survival rates for all TC were 80, 75 and 75%, respectively. In the first 5 years after diagnosis the crude death ratio was higher with the rise of age and for the follicular type and after 5 years for males and advanced disease. After inclusion of surgical treatment into the model, the estimates of the other death ratios did not change. Patients treated with 131I did better only during the first 5 years. Although the prognosis for TC patients treated in general hospitals in Southeastern Netherlands was similar to that found for patients treated in referral centres, concentration of treatment should be considered. PMID- 9849487 TI - Inhibition of human telomerase by a retrovirus expressing telomeric antisense RNA. AB - Human telomerase, the RNA-dependent DNA polymerase that adds TTAGGG repeats to chromosome ends, is selectively expressed in immortalised cells and most tumours, suggesting a potential role for telomerase inhibitors in cancer therapy. Replication-deficient retroviruses were used to determine whether mRNA containing UUAGGG, the complementary sequence to the template region of the hTR telomerase RNA, is sufficient to inhibit telomerase activity. Telomerase activities measured by the telomeric repeat amplification protocol (TRAP) assay in extracts prepared from immortalised mouse fibroblasts, human HeLa cells and human kidney carcinoma cells were inhibited by 75% or greater in 26 of 56 cell clones expressing UUAGGG. Telomerase activity was not inhibited by expression of mRNA containing a transposed sequence, GGGAUU. Telomerase activities in vivo were inferred from changes in cellular morphology, proliferation capacity, growth rate and measurement of the content of telomere DNA. Giant senescent-like cells emerged shortly after cloning mouse PA317 and human HeLa cells expressing UUAGGG. The fraction of giant cells varied from 100% at the fifth population doubling (PD) in one culture to 2-6% at 50 PD in several other cultures. Giant cells were absent in all parental cells and clones expressing GGGAUU. The average cellular content of telomere DNA was independent of telomerase activity over 50 PD. The results indicate that expression of RNA complementary to the template region of hTR is sufficient to inhibit telomerase in vitro and in vivo, but that the effect of inhibition on individual cells is highly variable. PMID- 9849489 TI - 8-chloroadenosine 3',5'-monophosphate (8-Cl-cAMP) selectively eliminates protein kinase A type I to induce growth inhibition in c-ras-transformed fibroblasts. AB - 8-Chloroadenosine 3',5'-monophosphate (8-Cl-cAMP), a site-selective cyclic adenosine 3',5'-monophosphate (cAMP) analogue, exhibits growth inhibition in a broad spectrum of cancer cell lines. We investigated the effect of 8-Cl-cAMP on c ras-transformed mouse fibroblasts (MP3/3T3) which were established by transfection of Balb3T3 cells (Balb3T3) with the point-mutated c-ras gene [G12- >V12]. 8-Cl-cAMP (2-5 microM) exerted over 80% growth inhibition by day 4 on MP3/3T3, while inhibiting parental Balb3T3 cell growth less than 40%. In order to distinguish the effect of 8-Cl-cAMP from that of 8-chloroadenosine (8-Cl adenosine), we examined the effect of 8-Cl-cAMP in serum-free medium. 8-Cl-cAMP demonstrated a potent growth inhibition of MP3/3T3 cells cultured in serum-free medium, suggesting that the growth inhibitory effect of 8-Cl-cAMP was not due to its hydrolysed product, 8-Cl-adenosine. In addition, both Balb3T3 and MP3/3T3 contained cAMP phosphodiesterases mainly composed of isozyme IV which has previously been reported to be insensitive towards the hydrolysis of 8-Cl-cAMP. Non-transformed Balb3T3 cells contained only type II cAMP-dependent protein kinase (PKA), whereas transformed MP3/3T3 exhibited a marked increase in type I PKA. The growth inhibition of MP3/3T3 by 8-Cl-cAMP accompanied almost complete elimination of type I PKA without affecting type II PKA. Moreover, 8-Cl-cAMP induced an arrest in the G0/G1-phase of the cell cycle in MP3/3T3. 8-Cl-adenosine had little or no effect on the cell cycle kinetics of MP3/3T3 cells. These results show that 8-Cl-cAMP is a novel cAMP analogue which selectively eliminates type I PKA to induce growth inhibition in transformed fibroblasts. PMID- 9849488 TI - Enhancement of chemotherapeutic drug toxicity to human tumour cells in vitro by a subset of non-steroidal anti-inflammatory drugs (NSAIDs). AB - The effect on cytotoxicity of combining a range of clinically important non steroidal anti-inflammatory drugs (NSAIDs) with a variety of chemotherapeutic drugs was examined in the human lung cancer cell lines DLKP, A549, COR L23P and COR L23R and in a human leukaemia line HL60/ADR. A specific group of NSAIDs (indomethacin, sulindac, tolmetin, acemetacin, zomepirac and mefenamic acid) all at non-toxic levels, significantly increased the cytotoxicity of the anthracyclines (doxorubicin, daunorubicin and epirubicin), as well as teniposide, VP-16 and vincristine, but not the other vinca alkaloids vinblastine and vinorelbine. A substantial number of other anticancer drugs, including methotrexate, 5-fluorouracil, cytarabine, hydroxyurea, chlorambucil, cyclophosphamide, cisplatin, carboplatin, mitoxantrone, actinomycin D, bleomycin, paclitaxel and camptothecin, were also tested, but displayed no synergy in combination with the NSAIDs. The synergistic effect was concentration dependent. The effect appears to be independent of the cyclo-oxygenase inhibitory ability of the NSAIDs, as (i) the synergistic combination could not be reversed by the addition of prostaglandins D2 or E2; (ii) sulindac sulphone, a metabolite of sulindac that does not inhibit the cyclooxygenase enzyme, was positive in the combination assay: and (iii) many NSAIDs known to be cyclo-oxygenase inhibitors, e.g. meclofenamic acid, diclofenac, naproxen, fenoprofen, phenylbutazone, flufenamic acid, flurbiprofen, ibuprofen and ketoprofen, were inactive in the combination assay. The enhancement of cytotoxicity was observed in a range of drug sensitive tumour cell lines, but did not occur in P-170-overexpressing multidrug resistant cell lines. However, in the HL60/ADR and COR L23R cell lines, in which multidrug resistance is due to overexpression of the multidrug resistance-associated protein MRP, a significant increase in cytotoxicity was observed in the presence of the active NSAIDs. Subsequent Western blot analysis of the drug sensitive parental cell lines, DLKP and A549, revealed that they also expressed MRP and reverse-transcription-polymerase chain reaction studies demonstrated that mRNA for MRP was present in both cell lines. It was found that the positive NSAIDs were among the more potent inhibitors of [3H]-LTC4 transport into inside-out plasma membrane vesicles prepared from MRP-expressing cells, of doxorubicin efflux from preloaded cells and of glutathione-S-transferase activity. The NSAIDs did not enhance cellular sensitivity to radiation. The combination of specific NSAIDs with anticancer drugs reported here may have potential clinical applications, especially in the circumvention of MRP-mediated multidrug resistance. PMID- 9849490 TI - bcl-2 expression is reciprocal to p53 and c-myc expression in metastatic human colorectal cancer. AB - Apoptosis (programmed cell death) inhibition may be an important mechanism by which gastrointestinal mucosal cells containing damaged DNA evade normal clearance mechanisms and grow to become invasive tumours. Since bcl-2 is an apoptosis inhibitor, bcl-2 mRNA expression was measured in 21 metastases of colorectal cancer using reverse transcription-polymerase chain reaction analysis. The mean bcl-2 mRNA expression (0.45 U, P < 0.0001) was lower than that of normal mucosal controls (= 1 U). p53 expression was inversely correlated with bcl-2 expression (P = 0.021) in 19 evaluable samples, and in tumours where p53 expression was over twice that of normal colonic mucosal values, bcl-2 mRNA was significantly decreased (mean 0.30, P = 0.0052). c-myc was also inversely correlated with bcl-2 expression (P = 0.025). Decreased bcl-2 expression in metastatic colorectal cancer may be partly due to allelic loss, given the proximity of bcl-2 to the frequently deleted DCC gene on chromosome 18q. However, the inverse correlation to p53/c-myc suggests an active downregulation of bcl-2, possibly following delegation of its apoptosis inhibiting role to other genes. PMID- 9849491 TI - Design of a novel oral fluoropyrimidine carbamate, capecitabine, which generates 5-fluorouracil selectively in tumours by enzymes concentrated in human liver and cancer tissue. AB - Capecitabine (N4-pentyloxycarbonyl-5'-deoxy-5-fluorocytidine) is a novel oral fluoropyrimidine carbamate, which is converted to 5-fluorouracil (5-FU) selectively in tumours through a cascade of three enzymes. The present study investigated tissue localisation of the three enzymes in humans, which was helpful for us to design the compound. Carboxylesterase was almost exclusively located in the liver and hepatoma, but not in other tumours and normal tissue adjacent to the tumours. Cytidine (Cyd) deaminase was located in high concentrations in the liver and various types of solid tumours. Finally, thymidine phosphorylase (dThdPase) was also more concentrated in various types of tumour tissues than in normal tissues. These unique tissue localisation patterns enabled us to design capecitabine. Oral capecitabine would pass intact through the intestinal tract, but would be converted first by carboxylesterase to 5' deoxy-5-fluorocytidine (5'-dFCyd) in the liver, then by Cyd deaminase to 5'-deoxy 5-fluorouridine (5'-dFUrd) in the liver and tumour tissues and finally by dThdPase to 5-FU in tumours. In cultures of human cancer cell lines, the highest level of cytotoxicity was shown by 5-FU itself, followed by 5'-dFUrd. Capecitabine and 5'-dFCyd had weak cytotoxic activity only at high concentrations. The cytotoxicity of the intermediate metabolites 5'-dFCyd and 5' dFCyd was suppressed by inhibitors of Cyd deaminase and dThdPase, respectively, indicating that these metabolites become effective only after their conversion to 5-FU. Capecitabine, which is finally converted to 5-FU by dThdPase in tumours, should be much safer and more effective than 5-FU, and this was indeed the case in the HCT116 human colon cancer and the MX-1 breast cancer xenograft models. PMID- 9849492 TI - Antitumour activity of the novel immune modulator 5,6-dimethylxanthenone-4-acetic acid (DMXAA) in mice lacking the interferon-gamma receptor. AB - 5,6-Dimethylxanthenone-4-acetic acid (DMXAA), a novel antitumour agent currently undergoing clinical evaluation, appears to mediate its antitumour effects through immune modulation and the production of cytokines. We used mice with a targeted disruption of the interferon-gamma (IFN-gamma) receptor gene as a model to evaluate the role of the host response to IFN-gamma in the antitumour action of DMXAA on colon 38 tumours. A feature of the results was that while DMXAA treatment induced both IFN-gamma and tumour necrosis factor (TNF) in serum, the increase was > 20-fold higher in IFN-gamma R0/0 mice than in wild-type mice. In contrast, mRNA levels for IFN-gamma and TNF were similar in the two mouse strains, suggesting that the concentrations of these cytokines were controlled by a post-transcriptional mechanism. Serum nitrate levels, used as a measure of nitric oxide production, were increased by DMXAA, but to a similar extent in both strains of mice. Complete regressions of colon 38 tumours were obtained in response to DMXAA in the knockout mice, although the dose required for 100% cure was higher and the reduction in tumour volume occurred more slowly than in the wild-type counterparts. The results demonstrate that the host response to IFN gamma is not essential for an anti-tumour response. Similar results were obtained in mice that were immunosuppressed by treatment with cyclosporin A before treatment with DMXAA. The results are consistent with the concept that the antitumour activity of DMXAA involves complex immunomodulation, probably with significant redundancy in contributing cytokines. PMID- 9849493 TI - Phase I study of a weekly 1 h infusion of paclitaxel in patients with unresectable hepatocellular carcinoma. AB - The majority of patients with hepatocellular carcinoma will develop either unresectable or metastatic disease and, therefore, are candidates for systemic chemotherapy. Only a few chemotherapeutic agents have shown documented activity in the treatment of advanced hepatocellular carcinoma and there is clearly a need for the evaluation of new active drugs. Therefore, we performed a phase I trial with a weekly schedule of paclitaxel in patients with advanced hepatocellular carcinoma. 16 patients with documented progression of unresectable hepatocellular carcinoma were included. After premedication, paclitaxel was given as a 1 h infusion on days 1, 8, 15, 22, 29 and 36 representing one treatment cycle. The cycle was repeated every 50 days. The starting dose was 70 mg/m2 and the doses were escalated in steps of 10 mg/m2/week. A minimum of 3 patients were treated at each dose level. All treatment was given on an out-patient basis. Dose-limiting toxicity was reached at a dose of 100 mg/m2/week with 2 of 6 patients treated at that dose level having WHO grade 4 neutropenia. Other toxic side-effects were only mild. 1 partial response and 9 cases with disease stabilisation were observed in 16 patients with initially progressive disease. We, therefore, conclude that the recommended dose for a further phase II trial in patients with hepatocellular carcinoma is 90 mg/m2/week. These data indicate that paclitaxel given at this dose and schedule might have activity in hepatocellular carcinoma and further investigation in phase II trials is warranted. PMID- 9849495 TI - Long lasting, grade IV, orthostatic hypotension after a single cycle combination chemotherapy with paclitaxel and cisplatin. PMID- 9849494 TI - Effects of ondansetron on gastrointestinal symptoms in carcinoid syndrome. AB - The effect of short-term treatment with the highly selective serotonin receptor antagonist ondansetron on symptoms and gastric emptying in 11 carcinoid patients was studied. Diarrhoea improved in 6 of 6 patients, nausea in 3 of 4 patients. Flushing was not affected. The rate of gastric emptying increased during ondansetron treatment (P = 0.08). No changes in serotonin in platelets and urinary excretion of 5-hydroxyindoleacetic acid were found. It is concluded that ondansetron can improve gastrointestinal symptoms in carcinoid patients and possibly slows gastric emptying. PMID- 9849496 TI - Palliative effect of gemcitabine in osteosarcoma resistant to standard chemotherapy. PMID- 9849497 TI - Severe anaphylactic reactions to oxaliplatin. PMID- 9849499 TI - Technology. Hooray, we're average! PMID- 9849498 TI - Vascular endothelial cell growth factor (VEGF) serum concentrations change according to the phase of the menstrual cycle. PMID- 9849500 TI - Public health. Altered stats. PMID- 9849501 TI - Public health. AIDS deaths drop. PMID- 9849502 TI - Gateway to good health. System is a leader in helping improve coverage for St. Louis's poor by 2004. PMID- 9849503 TI - Unhappy landings. Health care feels the fallout from Washington's 1997 balanced budget. AB - Health care is already feeling the fallout from the 1997 balanced budget pact. And the accord's $100 billion in Medicare cuts, spread over five years, are only beginning to take effect. H&HN looks at two especially hard-hit sectors: home health agencies and rural hospitals. PMID- 9849504 TI - All the hazards of home. PMID- 9849505 TI - A stormy season for rurals. PMID- 9849506 TI - Melatonin for the masses. Antiaging entrepreneurs peddle therapies that promise to cheat the clock. AB - The come-ons for a new wave of antiaging clinics may be over the top, but they speak to boomer anxieties about flagging energy levels and flabby midsections. And they just may be the next hot trend in health care. PMID- 9849507 TI - Outsourcing loses its 'MO'. Our annual survey points to a plateau for most contract services. AB - Outsourcing remains popular with health care executives hungry to control costs. But our annual survey shows that its momentum is stalling. Most respondents don't expect their contracting of business, support, and clinical services to change much over the next two years. PMID- 9849508 TI - Collective clout. Share the health. PMID- 9849509 TI - Count up 2000. The bug eating your budget. PMID- 9849510 TI - Dollars for dialysis. The height of helping out. PMID- 9849511 TI - Waste watch. Home's where the harm is. PMID- 9849512 TI - Digital revolution. The pain of going paperless. PMID- 9849513 TI - Ignoring the wishes and needs of an ill child cannot be right. PMID- 9849514 TI - Listening to children: have we gone too far (or not far enough)? PMID- 9849515 TI - The long-term course of mastalgia. AB - Mastalgia is a common condition in women of reproductive years. We have assessed the long-term course in patients with severe mastalgia by distributing a postal questionnaire to 212 patients previously studied in 1983 who had attended the mastalgia clinic at the University Hospital of Wales, Cardiff. 175 patients (83%) responded, with an original diagnosis of cyclical mastalgia (CM) in 120 and non cyclical mastalgia (NCM) in 55. The median age of onset of breast pain was 36 years (range 12-63 years). The average duration of pain was long (median 12 years), especially if it started in the second or third decade of life. Pain persisted in 68 (57%) of CM and 35 (64%) of NCM patients. In CM patients resolution was commonly associated with a 'hormonal' event, notably the menopause; in NCM patients it more often seemed to be spontaneous. Severe mastalgia ran a chronic relapsing course often requiring repeated drug treatments. PMID- 9849516 TI - Access to complementary medicine in general practice: survey in one UK health authority. AB - Complementary therapy (CT) has become increasingly popular with the general public and interest from the health professions has been rising. There has been no study focusing on the pattern of availability of CT within urban and inner city general practice. We aimed to describe the prevalence and pattern of access to complementary therapy in this setting, identifying the characteristics of practices offering CT and the perceived barriers to service provision. We sent a postal questionnaire to all 254 general practices on the Birmingham Family Health Services Authority list. Practices were asked whether they offered any access to CTs, how services were organized and which therapies were available and to identify any barriers to provision. 175 practices (68.9%) responded. Half of the practices offered access to CT. Of these, half offered an in-house service, usually provided by the doctor (81.8%). Of GPs practising therapies themselves, 58% began in or after 1990. Seventeen separate therapies were offered, most commonly acupuncture, osteopathy, chiropractic, hypnotherapy and homoeopathy. Practices significantly more likely to offer access to CT were of larger list size and training or teaching practices. They were equally likely to be fundholders or non-fundholders. Practices offering an in-house service tended to be fundholding, training and of larger list size. Finance was perceived as the major barrier. In the area studied, many patients now have some access to CT within primary care, often within their own practice. In the main, therapies offered are the 'medically acceptable face' of complementary medicine. PMID- 9849517 TI - A rude awakening: post-take ward round in the department of medicine for the elderly. AB - This paper describes the post-take ward round of a department of medicine for the elderly (DME), to portray the nature of the medical admissions and their immediate management. The data concern the patients seen by one consultant in 28 such ward rounds during the last four months of 1997, in a teaching hospital where the DME is separate from the department of general internal medicine. 254 patients were seen, 107 men and 147 women, with an average age of 82.4 years (range 73-102). The decisions taken included diagnosis, further investigations, treatment, referral, discharge, and resuscitation status. Very few admissions were judged inappropriate, particularly among the majority referred by general practitioners. 101 patients were thought suitable for transfer to the department of general internal medicine, 109 definitely unsuitable. These findings support the view that, if medical beds are to be freed, the initiative must come from facilitating discharge rather than curtailing admission. Generalists are needed to sort and manage these patients. In the UK, these will often be general internal medicine consultant geriatricians, while the younger patients are seen by consultants practising general internal medicine in addition to one of the specialties. Sizeable numbers of these consultants are needed if the post-take ward round is to be efficient and not conflict with their fixed commitments. PMID- 9849518 TI - Hormone replacement therapy and cardioprotection in the elderly. PMID- 9849519 TI - The control of breathing with reference to congenital central hypoventilation syndrome. PMID- 9849520 TI - In the Darwin family tradition: another look at Charles Darwin's ill health. PMID- 9849521 TI - Treatment of orofacial and ileo-colonic Crohn's disease with total enteral nutrition. PMID- 9849522 TI - Nipple discharge in a teenager. PMID- 9849523 TI - Oedeme bleu. PMID- 9849524 TI - Acute aortic thrombosis despite anticoagulant therapy in idiopathic hypereosinophilic syndrome. PMID- 9849525 TI - Mass in the neck after whiplash injury. PMID- 9849526 TI - Duodenocolic fistula causing rectal bleeding. PMID- 9849527 TI - A veterinary President. Interview by Robin Fox. AB - On 21 July, Lord Soulsby of Swaffham Prior took office as President of the Royal Society of Medicine. He qualified mrcvs from Edinburgh in 1948 and held lectureships in Bristol and Cambridge before appointment as Professor of Parasitology in the University of Pennsylvania in 1964. There he stayed for fourteen years, returning to Cambridge in 1978 as Professor of Animal Pathology (now Emeritus). His work as a parasitologist has taken him to the USSR, Nigeria, India, Australia, South America, China and numerous countries of Europe. Earlier presidencies have included the Royal College of Veterinary Surgeons, the World Association for the Advancement of Parasitology, the Cambridge Society for Comparative Medicine, and the Comparative Medicine Section of the RSM (1993-95); he is Patron of the Fund for Replacement of Animals in Medical Experiments. He has been a consultant to international bodies including WHO, the UN Development Programme, FAO, and the International Atomic Energy Agency. Created a life peer in 1990 (now on the Opposition benches), he chaired a Select Committee on antibiotic resistance whose report appeared earlier this year. He is interviewed here by Robin Fox. PMID- 9849528 TI - Haemophilia: does the future lie in replacement therapy or auto-supply? PMID- 9849529 TI - Cremation certificates. PMID- 9849530 TI - Autosurgery, 1816. PMID- 9849532 TI - Prostate cancer screening. PMID- 9849533 TI - The Bournewood case. PMID- 9849534 TI - Ibn Al-Nafis. PMID- 9849535 TI - Arterial thrombosis after a long-haul flight. PMID- 9849536 TI - Neurotoxicity in young adults 20 years after childhood exposure to lead: the Bunker Hill experience. AB - OBJECTIVES: An epidemiological study of young adults was conducted to determine whether environmental exposure to lead during childhood was associated with current adverse neurobehavioural effects. METHODS: The exposed group consisted of 281 young adults who had been exposed environmentally to lead as children and the unexposed referent group consisted of 287 age and sex frequency matched subjects. Information on demographics, past and current health, and past exposures to neurotoxicants, and responses to the Swedish Q16 questionnaire were collected by interview. Standard neurobehavioural and neurophysiological tests were administered by computer or trained technicians. K x ray fluorescence was used to estimate tibial bone lead concentrations among the exposed and unexposed groups. Associations were examined between the exposed group and referents and tibial bone lead concentration and the neurobehavioural and neurophysiological outcomes of interest. RESULTS: Among the measures of peripheral nerve function, after controlling for confounders, sural sensory nerve evoked response amplitude, peroneal motor nerve compound motor action potential amplitude, vibrotactile thresholds of fingers and toes, and standing steadiness were significantly associated with exposure group. Among the neurobehavioural tests, hand-eye coordination, simple reaction time latency, trails B latency, symbol digit latency, serial digit, and learning error score were also significantly associated with exposure group after controlling for confounders. Exposed subjects had significantly more neuropsychiatric symptoms than the referents. Associations between tibial bone lead concentration and scores for vocabulary, vibrotactile thresholds of the fingers, and vibrotactile thresholds of the toes approached significance. CONCLUSIONS: Significant adverse central and peripheral neurological effects were found in a group of young adults 20 years after childhood environmental exposure to lead when compared with non-exposed controls. The absence of a significant association between neurological outcomes and tibial bone lead concentration, and the presence of significant associations between neurological outcomes and exposure group may be due to either the magnitude of measurement uncertainty in K x ray films relative to the actual tibial bone lead concentration in these young non-occupationally exposed subjects, or uncontrolled confounding of the exposure group. PMID- 9849537 TI - Leukaemia, lymphoma, and multiple myeloma in seamen on tankers. AB - OBJECTIVES: To investigate the risk of lymphatic and haematopoietic malignancies in deck crew on tankers exposed to cargo vapours. METHODS: The study design was as a nested case-referent study in two cohorts of male Swedish seamen 20-64 years of age at the national census 1960 (n 13,449) and 1970 (n 11,290), respectively. Cases were detected by record linkage with the Swedish Cancer Register 1961-79 and 1971-87, respectively. For each case, three to five age matched referents from the population were selected. Exposure was assessed from data in the Swedish Registry of Seamen and from a register of Swedish ships. RESULTS: Seamen in the 1970 cohort, who had been exposed to cargo vapours for at least one month on chemical or product tankers, had an increased risk of lymphatic and haematopoietic malignancies (Mantel-Haenszel odds ratio (OR) 2.6, 95% confidence interval (95% CI) 1.1 to 5.9)) with a significant exposure-response relation (conditional logistic regression analysis, p = 0.04). The ORs were increased for both lymphoma (3.2), multiple myeloma (4.0), and leukaemia (1.6), but the increase was only significant for non-Hodgkin's lymphoma (OR 3.3, 95% CI 1.1 to 10.6). There were no significantly increased risks for the 1960 cohort or for seamen exposed only on crude oil tankers, but these groups had few exposed cases and low cumulative exposure to benzene and other light petroleum products. CONCLUSIONS: Seamen exposed to cargo vapours from gasoline and other light petroleum products on chemical or product tankers had an increased incidence of lymphatic and haematopoietic malignancies. One possible cause is exposure to benzene during loading, unloading, and tank cleaning operations. PMID- 9849538 TI - Agricultural use of DDT and risk of non-Hodgkin's lymphoma: pooled analysis of three case-control studies in the United States. AB - OBJECTIVES: The objective of this pooled analysis was to examine whether exposure to DDT was associated with the risk of non-Hodgkin's lymphoma among male farmers. METHODS: Data from three case-control studies from four midwestern states in the United States (Nebraska, Iowa, Minnesota, Kansas) were pooled to carry out analyses of 993 cases and 2918 controls. Information on use of agricultural pesticides and other risk factors was based on interviews. Non-farmers (people who had never lived or worked on a farm) were used as a reference category. RESULTS: There were 161 cases and 340 controls who reported use of DDT on animals or crops, or on both, yielding an odds ratio (OR) of 1.2 (95% confidence intervals (95% CI) 1.0 to 1.6). Farmers who had used DDT for > or = 15 years had an OR of 1.5 (95% CI 1.0 to 2.3). Adjustment for respondent status and use of other pesticides resulted in slightly reduced ORs. Analyses by the number of days of use a year was limited to the Nebraska data. The most notable increase was found among farmers who used DDT for > or = 5 days a year (OR 2.6, 95% CI 1.1 to 5.9); however, additional adjustment for use of organophosphates, phenoxyacetic acids, and the individual pesticides lindane, malathion, and atrazine reduced the ORs to 1.0, 0.9, 1.1, 1.6, and 1.9 respectively. CONCLUSIONS: No strong consistent evidence was found for an association between exposure to DDT and risk of non-Hodgkin's lymphoma. It seems that the excess risk initially found may be explained by use of other pesticides. PMID- 9849539 TI - Increased morbidity odds ratio of primary liver cancer and cirrhosis of the liver among vinyl chloride monomer workers. AB - OBJECTIVES: To determine if there is an increased risk of admission to hospital for various diseases among vinyl chloride monomer (VCM) workers. METHODS: 2224 workers with occupational exposure to VCM were identified for occurrence of disease based on a search of hospital computer files on labour insurance. These data were compared with those of workers manufacturing optical equipment and motorcycles from 1 January 1985 to 31 March 1994. Cardiovascular and cerebrovascular diseases were used as reference diseases, and the age adjusted morbidity odds ratio (MOR) was calculated. RESULTS: A significantly increased risk of admission to hospital among VCM workers due to primary liver cancer (MOR 4.5-6.5), cirrhosis of the liver (MOR 1.7-2.1), and other chronic diseases (MOR 1.5-2.0) was found. There were eight cases of primary liver cancer, all with heavy previous exposure to VCM. Another four cases of hepatoma in polyvinyl chloride (PVC) workers were found in the death registry. Ten out of 11 cases of hepatoma, with detailed medical information, were carriers of hepatitis B virus. The average latent period (20 years) was not different from other studies. Alternative agents of primary liver cancer were largely ruled out, suggesting that the combination of hepatitis B and VCM may lead to primary liver cancer. CONCLUSION: There is an increased risk of primary liver cancer in workers exposed to VCM, although the incomplete coverage of the Labor Insurance Bureau data warrants cautious interpretation of the results. Further study exploring the synergistic effects of VCM and hepatitis B is also indicated. PMID- 9849540 TI - Relations between occupational exposure to coal mine dusts, erythrocyte catalase and Cu++/Zn++ superoxide dismutase activities, and the severity of coal workers' pneumoconiosis. AB - OBJECTIVES: To better understand the relations between occupational exposure, blood antioxidant enzyme activities, total plasma antioxidant concentration, and the severity of coal workers' pneumoconiosis (CWP). METHODS: Blood samples were obtained from miners without CWP exposed to low dust concentrations for > or = 4 years at the time of the study (n = 105), or exposed to high dust concentrations for > or = 14 years at the time of the study (n = 58), and from retired miners with CWP (n = 19). Miners without CWP were classified into three subgroups according to their estimated cumulative exposure to dust. Chest x ray films were obtained for each miner. Miners were classified in five subgroups according to their International Labour Organisation (ILO) profusion grades. Univariate tests were completed by multiple linear regression analyses. RESULTS: The estimated cumulative exposure to dust was strongly positively related to erythrocyte catalase activity and strongly negatively related to Cu++/Zn++ SOD activity only in miners exposed to high dust concentrations for > or = 14 years at the time of the study (F tests p = 0.006 and p = 0.004 respectively). Moreover, catalase activity was strongly related to the severity of CWP expressed as five subgroups of ILO profusion grades (F test p = 0.003); the greatest difference in the mean values was found between the group of 1/1 to 1/2 ILO profusion grades and the group of 2/1 to 3/3 ILO profusion grades. CONCLUSION: These results are in good agreement with the hypothesis that production of reactive oxygen species may be an important event in the exposure to coal mine dusts and the severity of CWP. Erythrocyte catalase and Cu++/Zn++ SOD activities are more closely related to recent exposure to high dust concentrations than to cumulative exposure, and could be considered as biological markers of exposure rather than as markers of early adverse biological effect. PMID- 9849541 TI - Association between hospital emergency visits for asthma and air pollution in Valencia, Spain. AB - OBJECTIVE: To assess the short term effect of concentrations of black smoke, sulphur dioxide (SO2), nitrogen dioxide (NO2) and ozone (O3) in ambient air on emergency room visits for asthma in the city of Valencia, Spain during the period 1994-5. METHODS: Ecological study with time series data and application of Poisson regression. Associations between number of daily emergency visits in a city's hospital and concentrations of air pollutants were analysed taking into account potential confounding factors by the standardised protocol of the air pollution and health: a European approach (APHEA) project. RESULTS: Mean (range) daily number of emergency room visits for asthma was 1 (0-5). Concentrations of all pollutants studied remained within current air quality standards. The association between an increase of 10 micrograms/m3 in ambient air pollution and asthma, measured as a relative risk (RR) of emergency visits, was significant for NO2 24 hour mean (lag 0, RR 1.076, 95% confidence interval (95% CI) 1.020 to 1.134), NO2 hour maximum (lag 0, RR 1.037, 95% CI 1.008 to 1.066), and O3 hour maximum (lag 1, RR 1.063, CI 95% 1.014 to 1.114). The association was not significant for SO2 or for black smoke during the period analysed. The effects were not significantly different for the time of year, cold months (November to April), or warm months (May to October). CONCLUSIONS: Current concentrations of ambient air pollution in Valencia are significantly associated with emergency room visits for asthma. This association is high and more consistent for NO2 and O3 than for particulate matter and SO2 (classic pollutants). PMID- 9849542 TI - Job strain, social support at work, and incidence of myocardial infarction. AB - OBJECTIVES: The combination of high psychological job demands and low decision latitude (high job strain) has been associated with an increased risk of coronary heart disease. It has been proposed that this may also be the case for low social support at work. The aim of this study was to analyse the relations between these psychosocial factors and incidence of myocardial infarction. METHODS: Associations between psychosocial work characteristics and incidence of myocardial infarction was investigated through a population based case-control study. The study base comprised employed men and women in five Swedish counties during the years 1976-84. Cases of first myocardial infarction were identified from hospital discharge registers and death records from outside hospital, controls were selected through a random sample, and psychosocial work environment was assessed through a job exposure matrix on the basis of the occupation in the 1970 and 1975 censuses. RESULTS: An increased incidence of myocardial infarction was found for men and women in occupations characterised by low decision latitude. For men this increase was seen primarily in combination with high psychological demands (high job strain) and low social support at work. Younger men (30-54 years of age) in occupations with both high job strain and low social support at work had a relative risk of 1.79 (95% confidence interval (95% CI) 1.22 to 2.65) compared with subjects in low strain and high social support jobs after controlling for age, county of residence, and socioeconomic group. CONCLUSIONS: Our results indicate that jobs characterised by low decision latitude, high job strain, or low social support at work may be associated with an increased risk of acute myocardial infarction. If these associations are causal they may be of substantial importance from the point of view of workers' health. PMID- 9849543 TI - Comparison between two programmes for reducing the levels of risk indicators of heart diseases among male professional drivers. AB - OBJECTIVES: To compare two programmes for reducing the levels of risk indicators of heart diseases among professional drivers. The programmes were focused on changes of lifestyle. The aim of the programmes was to initiate and motivate a process of change within the driver, which in the long term should lead to permanent and sound health habits. One programme was based on health profile assessment and the other was a health examination. METHODS: Altogether, 102 subjects were investigated (51 allocated to an intervention group and 51 to a reference group). The programme in the intervention group (health profile assessment) was based on revelatory communication, adjusted to the driver and contained individual and group activities. The reference group went through a health examination. In both groups blood pressure, serum lipid concentrations, body mass index, and estimated maximal oxygen uptake were measured and the lifestyle habits were surveyed by questionnaires at the start and at follow ups of 6 and 18 months. RESULTS: The results showed that in the intervention group the maximal oxygen uptake increased, as did exercise habits and the intention to practice good dietary habits. Variable working hours was the most common obstacle to change a health habit. In the reference group the maximal oxygen uptake increased and the concentration of serum total cholesterol and the number of people who perceived stress and loneliness decreased. CONCLUSIONS: Both the health profile assessment and the health examination had an effect on the levels of some risk indicators of heart diseases. Both programmes turned out to be useful because of high participation during the entire period and a generally positive attitude among the subjects. PMID- 9849544 TI - Assessment of bibliographic databases performance in information retrieval for occupational and environmental toxicology. AB - OBJECTIVE: To determine the efficiency of the major bibliographic databases by assessing the percentage of references among the total literature available that can be retrieved from each database. We also evaluated the best database combinations to carry out an exhaustive search. METHODS: BIOSIS, EMBASE, MEDLINE, NIOSH-TIC, and TOXLINE were searched on two topics: allergy to latex and asbestos and mesothelioma, in the title, abstract, or keywords (textwords). This search was performed for the years 1994 and 1995. All the records were classified by journal and author's name and were verified for each record whether or not it was indexed in each database. Statistical analysis was performed with chi 2 test. RESULTS: 777 articles in 510 issues were found. The efficiency of each database (percentage of articles recovered) and of combinations varied between 11% and 63% for one database and between 42% and 86% for a combination of two databases. The reasons why these differences exist between databases, and within a database, between two different subjects or two different years are reported. CONCLUSION: Firstly, it is not advisable to assert that a bibliography is complete when only one database is searched. Secondly, the efficiency of the databases may be quite different. Finally, it is suggested that the best way to be as exhaustive as possible is to search two or more databases-for example, in EMBASE and TOXLINE, or to a lesser extent EMBASE and MEDLINE. This seems to be the best compromise solution between time consumed for searching and efficiency. PMID- 9849545 TI - Risk of hepatitis A infection in sewage workers. AB - OBJECTIVE: To evaluate the risk of hepatitis A virus (HAV) infection among sewage workers from occupational exposure to raw sewage. METHODS: An analytical cross sectional study of 241 company employees with possible occupational exposure to sewage in a large water and sewerage company was carried out. Previous exposure to hepatitis A virus infection was assessed, as were its associations with possible risk factors. RESULTS: Frequent occupational exposure to raw sewage was a significant risk factor for HAV infection, independently of other known risk factors (odds ratio 3.73, 95% confidence interval 1.48 to 9.37). Of 50 employees who reported occupational exposure to raw sewage most of the time, 30 (60%) had had HAV infection. CONCLUSION: Employees who are likely to be at risk of frequent exposure should have their immunity ensured. The salivary assay for IgG anti-HAV used in the study was highly specific and would be suitable for prevaccination testing of older employees, who are more likely to be immune. PMID- 9849546 TI - Exposure to blood borne viruses and the hepatitis B vaccination status among healthcare workers in inner London. AB - Occupational exposure to blood borne viruses was examined during one year at a London teaching hospital. A total of 236 incidents occurred of which 83% were related to sharps, 32% were clearly avoidable, and 7% involved an infected source patient. Overall uptake of hepatitis B vaccine was 78% but it was particularly low in paramedical (70%) and domestic staff (45%). Continued effort needs to be applied to improve uptake of hepatitis B vaccine and to maintain high standards of control of infection. PMID- 9849547 TI - Chinese herbal medicine, sibship, and blood lead in children. AB - OBJECTIVES: Risk factors for increased blood lead concentration (BPb) has been investigated. However, the effect of sibship and Chinese herbal medicine on BPb has not been systematically studied. In this study BPb data from voluntary testing was used to determine if Chinese herbal medicine and sibship were associated with BPb. METHODS: 319 children aged 1-7 were tested for BPb. Meanwhile, parents were interviewed to obtain information including consumption of Chinese herbal medicine, living environment, lifestyle, and sibship of the children tested. RESULTS: The mean (SD) BPb of 319 preschool children was 4.4 (2.4) micrograms/dl. The consumption of Ba-baw-san (a Chinese herbal medicine) was significantly associated with increased BPb in children (p = 0.038). Further multivariate regression analysis of BPb in 50 pairs of siblings showed the factors of being brothers explained 75% of variation for BPb, and being sisters and brother-sister explained 51% and 41% of variation respectively. CONCLUSION: Chinese herbal medicine and children's play patterns within the family expressed in different types of sibship are the main determinants of low concentrations of BPb in preschool children of Taiwan. PMID- 9849548 TI - Prevalence of occupational lung disease among Botswana men formerly employed in the South African mining industry. PMID- 9849549 TI - Epidemics of syphilis in the newly independent states of the former Soviet Union. PMID- 9849550 TI - Sexually transmitted diseases in sexually abused children: medical and legal implications. AB - Sexually transmitted diseases (STDs) may be transmitted during sexual assault. In children, the isolation of a sexually transmitted organism may be the first indication that abuse has occurred. Although the presence of a sexually transmissible agent from a child beyond the neonatal period is suggestive of sexual abuse, exceptions do exist. In this review I discuss the issues of the transmissibility and diagnosis of STDs in the context of child sexual abuse. Rectal or genital infection with Chlamydia trachomatis among young children may be the result of perinatally acquired infection and may persist for as long as 3 years. A major problem with chlamydia testing in the context of suspected sexual abuse in children has been the inappropriate use of non-culture tests. Although the new generation of nucleic acid amplification tests have shown high sensitivity and specificity with genital specimens from adults, data on use of these tests on any site in children are practically non-existent. Bacterial vaginosis (BV) has been identified among children who have been abused and among those who have not been abused. However, many of the methods used to diagnose BV in adults have not been evaluated in children. Recent studies of perinatal infection with human papillomavirus (HPV) have been inconclusive. HPV DNA has been detected at various sites in children who have not been abused. The relation to the development of clinically apparent genital warts is unclear. Although HIV can be acquired through sexual abuse in children, the exact risk to the child and which children should be screened is still controversial. PMID- 9849551 TI - Role of type specific herpes simplex virus serology in the diagnosis and management of genital herpes. AB - OBJECTIVES: To investigate the indications for the use of a type specific antibody test for herpes simplex virus in a department of genitourinary medicine in the United Kingdom. METHOD: Retrospective analysis of case records of 127 patients who accepted the test during a 20 month period. RESULTS/CONCLUSION: The test contributed to patient management in 79% of patients with recurrent genital ulceration of unknown cause. It was also useful for counselling a number of patients with initial episodes of disease and the asymptomatic partners of some patients when the partners were shown to possess antibodies specific to herpes simplex virus type 2. When evaluating sexual partners, the test was difficult to interpret if an isolate from the index case had not been typed. Access to viral typing may therefore be a greater priority than serological testing. As adverse psychological sequelae may follow the identification of an asymptomatic chronic infection, guidelines for the use of a type specific serological test are proposed. PMID- 9849552 TI - Presentation, pathology, and outcome of HIV associated renal disease in a specialist centre for HIV/AIDS. AB - OBJECTIVES: To describe the presentation, pathology, and outcome of biopsy proved renal disease in HIV infected patients at a central London HIV unit from 1992 to 1996. METHODS: Retrospective review of a computerised database and case notes to identify patients with renal disease confirmed by antemortem percutaneous renal biopsy or necropsy. RESULTS: 17 patients were identified, 13 had biopsy and four necropsy confirmed renal disease. Abnormalities included HIV associated nephropathy (HIVAN) in seven (41%) patients, membranous glomerulonephritis (GN) in four (23%), haemolytic uraemic syndrome (HUS) in two (12%), and interstitial nephritis, rhabdomyolysis, IgA nephropathy, and membranoproliferative GN in one patient each. Although renal disease was the first presentation of HIV disease in six (35%) patients the majority had advanced HIV disease (median CD4 count 40 x 10(6)/l). The commonest presentation was acute renal failure (ARF) in 10 (59%) patients, chronic renal failure (CRF) in five (29%), and proteinuria alone in two (12%). Although the majority of patients died during the study period (9/13) only three deaths were attributable to their renal disease. Survival ranged in those with HIVAN from 0 to 31 (median 10) months and, in those with membraneous GN from 1 to 46 (median 29) months. CONCLUSIONS: HIVAN was the commonest renal disease found in this group of patients; however, a variety of other pathologies were seen with variable outcomes. All cases of HIVAN were in patients of African or Afro-Caribbean origin and for the majority this was their first presentation of HIV disease. Nephrologists need to be aware of the possibility of HIV infection in patients presenting with renal disease. PMID- 9849553 TI - HIV infection among homosexual/bisexual males attending genitourinary clinics in Scotland. AB - BACKGROUND: Since 1991, unlinked anonymous HIV testing of homosexual/bisexual males attending genitourinary clinics in Edinburgh and Glasgow has been conducted and resulting prevalence data have been published annually. More detailed information which provides an understanding of what proportion of HIV infected men attending genitourinary clinics in central Scotland (i) remain undetected, (ii) acquire sexually transmitted infections following HIV diagnosis, and (iii) possibly become HIV infected either abroad or following sex with someone from abroad, is reported by the authors. METHODS: Unlinked anonymous HIV testing of syphilis serology specimens from homosexual/bisexual males attending genitourinary clinics during 1991-5. RESULTS: Of 3468 specimens tested, 165 (4.8%) were HIV positive. Thirty five per cent (57) of all HIV positive specimens were from men whose infection remained undetected following clinic attendance. Of the 80 attenders who knew themselves to be HIV positive before their clinic visit, 13 had clinical and/or laboratory evidence of a sexually transmitted infection. Men who had a sexual risk associated with America or who were American, had a 2.4-fold greater risk of being HIV infected than those with United Kingdom only connections. CONCLUSION: Increased efforts should be made to ensure that HIV infected men are diagnosed early after infection and do not engage in high risk sexual behaviour, and that all homosexual men are educated about the particular risks of acquiring HIV infection abroad. More effective interventions to prevent indigenous HIV transmissions need to be developed. PMID- 9849554 TI - Rapid assessment of sexually transmitted diseases in a sentinel population in Thailand: prevalence of chlamydial infection, gonorrhoea, and syphilis among pregnant women--1996. AB - OBJECTIVE: To determine the prevalence of sexually transmitted diseases (STDs) among pregnant women in Thailand, where case reporting suggests a marked decrease in STDs following a campaign promoting condom use during commercial sex. DESIGN: Cross sectional study of women at their first visit to the study hospitals' antenatal clinics in Chiang Rai (n = 500) and Bangkok (n = 521). METHODS: First catch urine specimens were tested for Chlamydia trachomatis and Neisseria gonorrhoeae using the Amplicor CT/NG polymerase chain reaction assay. Syphilis and HIV serological testing were performed in the study hospitals' laboratories. RESULTS: The prevalence of chlamydial infection was 5.7%, gonorrhoea 0.2%, and syphilis 0.5% (all VDRL or RPR titres were < or = 1:4). The prevalence of HIV infection was 7.1% in Chiang Rai and 2.9% in Bangkok. In a multivariate logistic regression analysis, chlamydial infection was associated with younger age and with higher gestational age at first antenatal clinic visit, but was not associated with marital status, gravidity, city of enrollment, or HIV infection status. CONCLUSIONS: There was a low prevalence of gonorrhoea and syphilis among these pregnant women in Thailand. Chlamydial infection was detected at a higher prevalence, especially among younger women and women registering later for antenatal care. Testing of pregnant women using easily collected urine specimens and a sensitive nucleic acid amplification assay is a feasible method of rapidly assessing chlamydial and gonococcal prevalence. PMID- 9849555 TI - Evaluation of two clinical protocols for the management of women with vaginal discharge in southern Thailand. AB - OBJECTIVES: (1) To compare the effectiveness of two clinical protocols for the management of vaginal discharge in the situations where no laboratory facilities are available but speculum examination is possible and where basic laboratory facilities are available. (2) To determine clinical and simple laboratory indicators for diagnosis of patients with vaginal discharge in the local setting. DESIGN: Alternate allocation of subjects to one of two management protocols. SUBJECTS: Women presenting to university gynaecology outpatients department with a complaint of vaginal discharge. METHODS: Subjects were alternately allocated management according to one of two protocols: one without (group A) and one with (group B) immediate access to results of basic laboratory tests. Full clinical assessment including speculum examination and microbiological assessment for infection with gonorrhoea, chlamydia, candida, trichomonas, and bacterial vaginosis was performed on all women. Follow up assessment of clinical and microbiological response was performed 1-2 weeks later. RESULTS: At initial assessment, both groups were similar in all respects except that more group B women had inflammation of the vulva. The prevalences of various conditions were: candidiasis 22%, bacterial vaginosis 38%, trichomoniasis 4%, chlamydia 4%, gonorrhoea 0.4%. There was no association between any demographic characteristic and diagnosis of cause of the discharge. Both protocols resulted in clinically and statistically significant improvements for women with candidiasis, bacterial vaginosis, and trichomoniasis. There were no clinically important differences in outcomes between the two protocols. The sensitivities and specificities of various indicators were: curd-like vaginal discharge for candidiasis, 72% and 100%; homogeneous vaginal discharge for bacterial vaginosis or trichomoniasis, 94% and 88%; absent or scanty lactobacilli for bacterial vaginosis, 99% and 68%; > 20% clue cells for bacterial vaginosis, 81% and 99%; visible endocervical mucopus for chlamydia or gonorrhoea, 36% and 86%; microscopic endocervical mucopus for chlamydia or gonorrhoea, 64% and 69%. CONCLUSIONS: Both protocols were equally effective in managing women with abnormal vaginal discharge. Simple clinical indicators for candidiasis, bacterial vaginosis, or trichomonas as in protocol A are sufficiently sensitive and specific for use in situations with no laboratory support. A modification to protocol A could increase detection of bacterial vaginosis at basic health service level. Further work is needed to identify appropriate indicators for infection with chlamydia or gonorrhoea. PMID- 9849556 TI - The supermarket for women's reproductive health: the burden of genital infections in a family planning clinic in Nairobi, Kenya. AB - OBJECTIVES: To study the burden of disease of reproductive tract infections (RTIs) and cervical dysplasia in women attending a family planning clinic in Nairobi, Kenya, and to assess the acceptability of integrating reproductive healthcare services into existing family planning facilities. METHODS: In a family planning clinic in Nairobi, Kenya, 520 women were enrolled in a study on RTI and cervical dysplasia. RESULTS: RTI pathogens were detected in over 20% of women, the majority being asymptomatic. HIV-1 testing was positive in 10.2%. The diagnosis of cervical dysplasia was made on 12% of the cytology smears (mild in 5.8%, moderate in 3.5%, severe in 1.2%), and 1.5% had invasive cervical cancer. The intervention of case detection of RTI and Papanicolaou smear taking was well received by clients and considered feasible by the staff. CONCLUSIONS: Early detection and treatment of potentially curable cervical lesions and RTI provide a unique opportunity to improve women's health. In Kenya, where the current contraceptive prevalence rate is 33%, family planning clinics are excellent sites to introduce health interventions. PMID- 9849557 TI - Auxotypes, serovars, and trends of antimicrobial resistance of Neisseria gonorrhoeae in Kigali, Rwanda (1985-93). AB - OBJECTIVE: To investigate the in vitro antimicrobial susceptibility and the auxotype/serovar distribution of Neisseria gonorrhoeae in Kigali, Rwanda, during 1985-93. METHODS: As part of a monitoring programme the in vitro susceptibility of 1604 isolates of N gonorrhoeae was determined by agar dilution. Auxo- and serotyping was performed on 1350 and 1313 isolates respectively. RESULTS: The prevalence of penicillinase producing N gonorrhoeae (PPNG) remained stable at a rate of 39% during 1985-91 and increased to 61% in 1992-3. Chromosomal resistance to penicillin was common among non-PPNG and resistance to thiamphenicol and tetracycline was common among both PPNG and non-PPNG. High level, plasmid mediated resistance to tetracycline (TRNG) was observed for the first time at the end of 1989 and increased from 2% of the isolates in 1990 to 50% by 1993. A trend for increasing resistance to norfloxacin and ofloxacin was observed during 1985 90 but disappeared in 1991-93. Five isolates with high level resistance to norfloxacin (MIC 2 mg/l) were observed in 1990. Resistance to trimethoprim sulphamethoxazole (TMP-SMZ) emerged at the end of 1990 and was observed among 10% of the isolates during 1991-3. All strains remained susceptible to ofloxacin, ciprofloxacin, spectinomycin, and ceftriaxone. Overall, 75% of the isolates were prototrophic or required proline for their growth and 62% belonged to serovars IA 6 and IB-1. The prevalence of serovar IB-4 increased strongly during the last 3 years of the study. CONCLUSION: Resistance to penicillin, thiamphenicol, and tetracycline was common in N gonorrhoeae during 1985-1993. The rapid spread of TRNG after 1989 and the steep increase of PPNG during 1992-3 were the most striking facts of the study period. The auxotype and serovar distribution was comparable with findings from other African countries. PMID- 9849558 TI - Antimicrobial susceptibility and plasmid profile of Neisseria gonorrhoeae in India (New Delhi). AB - OBJECTIVES: To determine the antibiotic susceptibility and plasmid profile of all Neisseria gonorrhoeae strains (PPNG and non-PPNG) isolated from May 1995 to March 1996 in Lok Nayak Hospital, New Delhi, India. METHODS: The agar plate dilution method was used to determine the minimum inhibitory concentration of five antimicrobials including norfloxacin and ceftriaxone which are most commonly used for treatment of gonorrhoea in Delhi. Isolates were screened for production of penicillinase by paper acidometric method and plasmid analysis of PPNG and non PPNG was carried out by agarose gel electrophoresis. RESULTS: 50 consecutive isolates of N gonorrhoeae were studied, 8% among them were found to be PPNG while 28% were highly resistant to tetracycline (TRNG). Reduced susceptibility to norfloxacin (MIC > or = 1 microgram/ml) was observed in 12% of all isolates. All PPNG harboured the 4.4 MDa beta lactamase plasmid along with the 25.2 MDa tetracycline resistance plasmid. Norfloxacin resistance (MIC > or = 1 microgram/ml) was present in 28.5% of TRNG but only in 5.5% of the other gonococcal isolates. CONCLUSIONS: Results of this study clearly demonstrate that antibiotic resistant gonococcal strains of different clones are frequently found in New Delhi. Continued surveillance of susceptibility to currently prescribed antimicrobials and epidemiological studies are essential to prevent treatment failures leading to further spread of resistant strains. PMID- 9849559 TI - Epidemiological study of Chlamydia trachomatis infection in pregnant women in Hungary. AB - A multicentre survey was carried out in order to determine the prevalence and risk factors of Chlamydia trachomatis infection in the pregnant population in Hungary. The nucleic acid hybridisation method (PACE 2 Gen-Probe) was applied for the examination of C trachomatis. The overall average prevalence of C trachomatis cases during an 18 month survey on 6161 pregnant women was 5.87%. There were significant differences in the proportions of chlamydial infection in the different survey centres, and also in the different age groups and the different family status groups. The perinatal mortality rate exhibited a significantly higher prevalence (8.52%) among C trachomatis positive than among negative patients (2.03%). In the anamnestic histories of C trachomatis infected patients, the frequency of premature uterine activity was 8.13%, in contrast with 5.18% in the non-infected group (p < 0.05). We suggest that all pregnant women be tested for C trachomatis infection. PMID- 9849561 TI - Genital herpes: Heisenberg revisited. AB - In the confirmation of recurrences of genital herpes, patient defined disease reactivation and virological data hold the scientific high ground. The influence of the psyche on recurrence rates and perception of recurrences has been largely neglected and marginalised up to the present, possibly because research work in that area has been and continues to be of poor calibre. However, neglected psychological variables may render otherwise relevant clinical trials uninterpretable. Psychological aspects of counselling before testing for serum herpes simplex type 2 antibodies are also discussed. PMID- 9849560 TI - Herpes simplex type 2 infection in a cohort aged 21 years. AB - OBJECTIVES: To measure the prevalence of HSV-2 antibodies in a birth cohort of 21 year old New Zealanders from whom detailed sexual histories had been obtained, and to assess the potential for HSV-2 serology in characterising a young adult population's risk for sexually transmitted diseases (STD). METHODS: Sera from 784 cohort members were tested using an indirect IgG enzyme linked immunoassay specific to the HSV-2 glycoprotein G. Positive results were confirmed by western blot. RESULTS: In all, 27 subjects were seropositive for HSV-2 (3.4%), only seven of whom gave a history of genital herpes (26%). Risk among females increased with lifetime number of partners, while risk for males increased with having a first partner who was aged 16 years or under. CONCLUSIONS: The seroprevalence of HSV-2 infection in this cohort was low, but similar to that seen in several other populations in this age group. HSV-2 seropositivity did not appear to be a sensitive marker for high risk sexual activity in this young population. This may be because a critical mass of HSV-2 carriers has not accumulated among potential partners by age 21 years. PMID- 9849562 TI - Multiple pyogenic granuloma of the penis. AB - A case of multiple pyogenic granuloma affecting the penis of a 28 year old man is reported. The lesions were arranged in a floret-like fashion around the inner aspect of the prepuce and developed after circumcision for congenital phimosis. Histopathological examination of sections from a biopsy specimen of the papillomatous growths revealed the findings of pyogenic granuloma. In this patient, the pathogenesis of the lesions is probably related to the failure in surgical wound repair that followed circumcision. Problems of clinical and histopathological differential diagnosis are discussed. PMID- 9849564 TI - Can a laboratory diagnosis of bacterial vaginosis be made from a transported high vaginal swab using anaerobic culture and microscopy of a wet preparation? PMID- 9849563 TI - How to assess an article on economic evaluation. PMID- 9849565 TI - Evaluation of a novel point of care test for antibodies to herpes simplex virus type 2. PMID- 9849566 TI - Emergence of fluoroquinolone resistant Neisseria gonorrhoeae in New Delhi, India. PMID- 9849567 TI - AIDS associated eosinophilic folliculitis which responded to both high dose co trimoxazole and low dose isotretinoin. PMID- 9849568 TI - Awareness of testicular tumours in genitourinary medicine clinics. PMID- 9849569 TI - Are race and ethnicity in STD analyses still of relevance? PMID- 9849571 TI - "Smegma stones". PMID- 9849570 TI - Evaluation of the commercial microparticle enzyme immunoassay. PMID- 9849572 TI - Peripheral blood B and T lymphocyte subsets. PMID- 9849573 TI - Journey beyond immunology. Regulation of receptor internalization by major histocompatibility complex class I (MHC-I) and effect of peptides derived from MHC-I. PMID- 9849574 TI - The PTEN, BAX, and IGFIIR genes are mutated in endometrial atypical hyperplasia. AB - To pursue the pathogenesis of endometrial carcinogenesis, we investigated microsatellite instability, mutations in the PTEN, TGF beta RII, IGFIIR, and BAX genes, and LOHs on 10q in 18 putative endometrial premalignant lesions (11 endometrial atypical hyperplasias (ATHs), 2 complex hyperplasias, and 5 simple hyperplasias) as well as 8 endometrial cancers (ECs). In the ATH cases, MSIs as well as LOHs at 10q were observed at frequencies similar to those in ECs. Mutations in PTEN, BAX, and IGFIIR were observed only in ATHs and ECs. These results suggest that (1) PTEN, BAX, and IGFIIR are already mutated in ATHs, and (2) ATH is one of the precursor lesions which could lead to EC. PMID- 9849575 TI - Exogenous epidermal growth factor exerts promoting action during the early phase of rat urinary bladder carcinogenesis. AB - Using the heterotopically transplanted rat urinary bladder (HTB) model that was developed in our laboratory, we examined the relationship between the duration of epidermal growth factor (EGF) treatment and acquisition of EGF-independence of urinary bladder tumors that were induced by EGF stimulation. After treatment with N-methyl-N-nitrosourea (MNU) (0.25 mg/0.5 ml of 0.9% NaCl once a week for 3 consecutive weeks), animals at week 3 received EGF [250 ng/0.5 ml phosphate buffered saline (PBS)] into the HTBs once a week for 20, 28, or 36 weeks. For examination of the effect of EGF withdrawal, one half of the rats received the vehicle (PBS) only beginning at week 23 or week 31 for 8 weeks. When animals were examined at week 23, the incidence and the mean number of tumors per bladder were low, irrespective of EGF treatment. In the bladders that had been exposed to EGF during the first 20 weeks after MNU administration, however, both the incidence and the mean number of tumors per bladder had increased significantly at week 31, regardless of whether or not EGF treatment was continued beyond week 23. Between weeks 31 and 39, EGF treatment demonstrated no effect; both the incidence of tumors and the mean number of tumors were the same as those at week 31. These results suggest that EGF exerts its promoting effect only during the early phase of MNU-initiated bladder carcinogenesis, but that its effect becomes manifest during the subsequent 8 weeks. EGF independence may be due to establishment of an autocrine growth-stimulatory mechanism in bladder tumors. PMID- 9849576 TI - The 1.5 GHz electromagnetic near-field used for cellular phones does not promote rat liver carcinogenesis in a medium-term liver bioassay. AB - We have recently established that local exposure to a 929.2 MHz electromagnetic near-field, used for cellular phones, does not promote rat liver carcinogenesis in a medium-term bioassay system. In the present study, a 1.439 GHz electromagnetic near-field (EMF), another microwave band employed for cellular phones in Japan, was similarly investigated. Time division multiple access (TDMA) signals for the Personal Digital Cellular (PDC) Japanese cellular telephone standard system were directed to rats through a quarter-wavelength monopole antenna. Numerical dosimetry showed that the peak SARs within the liver were 1.91 0.937 W/kg, while the whole-body average specific absorption rates (SARs) were 0.680-0.453 W/kg, when the time-averaged antenna radiation power was 0.33 W. Exposure was for 90 min a day, 5 days a week, over 6 weeks, to male F344 rats given a single dose of diethylnitrosamine (200 mg/kg, i.p.) 2 weeks previously. At week 3, all rats were subjected to a two-thirds partial hepatectomy. At week 8, the experiment was terminated and the animals were killed. Carcinogenic potential was scored by comparing the numbers and areas of the induced glutathione S-transferase placental form (GST-P)-positive foci in the livers of exposed (48) and sham-exposed rats (48). Despite increased serum levels of corticosterone, adrenocorticotropic hormone (ACTH) and melatonin, the numbers and the areas of GST-P-positive foci were not significantly altered by the exposure. These findings clearly indicated that local body exposure to a 1.439 GHz EMF, as in the case of a 929.2 MHz field, has no promoting effect on rat liver carcinogenesis in the present model. PMID- 9849577 TI - Prevention of N-methylnitrosourea-induced colon carcinogenesis in F344 rats by lycopene and tomato juice rich in lycopene. AB - Epidemiological studies have suggested a protective effect of lycopene and lycopene-rich tomatoes against various cancers. Here, the inhibition of colon carcinogenesis by lycopene and tomato juice was investigated. Seven-week-old female F344/NSlc rats received an intrarectal dose of 2 mg (experiment I) or 4 mg (experiment II) of N-methylnitrosourea 3 times a week for 3 weeks, and had free access to one of 4 drinking fluids: plain water (control group), 17 ppm lycopene water solution (Ly group), and diluted tomato juice containing 17 ppm (Tj group) or 3.4 ppm (tj group) lycopene, throughout the experiments. The colon cancer incidence at week 35 was significantly lower in the Tj group, but not in the Ly group, than in the control group: 21% and 33% vs. 54%, in experiment I (24 rats in each group). It was significantly lower in the Tj group than in the tj and control groups, 40% vs. 72% and 84%, in experiment II (25 rats in each group). An appreciable amount of lycopene (0.02 microgram/g) was detected in the colon mucosa of rats in the Tj group, but not in the tj group. The results suggest that tomato juice rich in lycopene may have a protective effect against colon carcinogenesis. PMID- 9849578 TI - Prevention of N-methylnitrosourea-induced colon tumorigenesis by ursodeoxycholic acid in F344 rats. AB - Bile acids are known to promote colon carcinogenesis. However, there is one study showing that ursodeoxycholic acid (UDCA) supplemented in the diet at the concentration of 0.4% prevented azoxymethane-induced rat colon tumorigenesis. The aim of our study was to explore the inhibitory effect of a much smaller dose of UDCA on colon carcinogenesis in rats. One hundred 7-week-old F344 rats were given 2 mg of N-methylnitrosourea 3 times a week for 3 weeks by intrarectal instillation, and were fed a 0% (control), 0.4% or 0.08% UDCA-supplemented diet for the next 27 weeks. All the rats were killed and examined for tumor development at week 30. The tumor incidence and number were significantly lower and smaller, respectively, in the UDCA-fed rats than in the control rats: 40% and 36% vs. 68%; 0.5 +/- 0.1 (mean +/- SEM) and 0.4 +/- 0.1 vs. 1.0 +/- 0.2. All the tumors were located in the distal half of the colon and were plaque-shaped or polypoid, being well-differentiated adenocarcinomas restricted to the mucosa or submucosa. Bile acids in the feces and the blood obtained at weeks 20 and 30, respectively, were analyzed by HPLC. A significant increase of UDCA was confirmed in both the feces and the blood of the UDCA-fed rats compared with the control rats. The results suggest that the continuous feeding of a small dose of UDCA may prevent colon carcinogenesis. PMID- 9849579 TI - Linkage mapping of the Bra, Brb and Brg genes for rat protein phosphatase 2A 55 kDa B-regulatory subunit isotypes. AB - We previously identified the rat Bra, Brb and Brg genes, which encode alpha, beta and gamma isotypes of the 55 kDa B-regulatory subunit of protein phosphatase 2A. Polymerase chain reaction-single strand conformation polymorphism analysis in the present study identified polymorphisms in Bra, Brb and Brg between the ACI and BUF, ZI and TM, and BN and WTC strains, respectively. Linkage analysis using mapping panels composed of F2 or back-crosses of these strains allowed Bra, Brb and Brg to be assigned to chromosomes 15, 18 and 14, respectively. Furthermore, it was revealed that Bra is located close to the Rb1 locus. Using polymorphism in Bra, loss of heterozygosity (LOH) was analyzed for rat mammary tumors induced in (SD x F344) F1 female rats by a food-borne carcinogen, 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine, and a typical mammary carcinogen, 7,12 dimethylbenz[a]anthracene. No LOH was detected at the Bra locus. PMID- 9849581 TI - Expression of cytochrome P450 3A4 in foveolar epithelium with intestinal metaplasia of the human stomach. AB - The expression of cytochrome P450 3A4 (CYP3A4) in the foveolar epithelium of the human stomach with intestinal metaplasia was studied using immunohistochemistry, western blotting and reverse transcription polymerase chain reaction (RT-PCR). CYP3A4 was immunohistochemically detected in the foveolar epithelium with intestinal metaplasia, but was not detected in foveolar epithelium without intestinal metaplasia, in the pyloric gland or in the fundic gland of the stomach. Western blotting and RT-PCR demonstrated that CYP3A4 protein and mRNA were expressed in the liver and pyloric gland mucosa with intestinal metaplasia, but not in the fundic gland mucosa without intestinal metaplasia. Possible roles of CYP expression in the gastric mucosa with intestinal metaplasia in human stomach carcinogenesis are briefly discussed. PMID- 9849580 TI - Microsatellite instability and frameshift mutations in the Bax gene in hereditary nonpolyposis colorectal carcinoma. AB - We studied microsatellite instability (MI) and bax gene abnormalities in colorectal carcinomas from 36 patients diagnosed as having hereditary nonpolyposis colorectal cancers (HNPCC) according to the clinical criteria (12 with confirmed HNPCC in group A and 24 at high risk of HNPCC in group B) and from 20 randomly selected patients with other colorectal cancers. MI was examined at 4 dinucleotide microsatellite loci and one mononucleotide locus. Frameshift mutations in the bax gene were detected by polymerase chain reaction-single strand conformation polymorphism analysis. MI was detected in 7 of the 12 patients in group A and 12 of the 24 in group B. Three MI patterns were identified: type 1, MI in both mono- and dinucleotide repeats; type 2, MI only in mononucleotide repeats and type 3, MI only in dinucleotide repeats. Most MI positive patients in group A showed type 1 MI, whereas in group B, 5 showed type 1, 3 showed type 2 and 4 showed type 3. Frameshift mutations in the bax gene correlated strongly with type 1 and type 2 MI. These results indicate that mutations of different DNA mismatch repair genes may cause several types of MI and result in several different clinical phenotypes of HNPCC. The bax gene may be one of the target genes which play a role in the tumorigenesis of HNPCC. PMID- 9849582 TI - Expression of CD44 variants and its association with survival in pancreatic cancer. AB - Since the CD44 variant 6(v6) molecule has been noted as a marker for tumor metastasis and prognosis in several tumors, we examined whether or not v6 is a useful marker for evaluating the prognosis of pancreatic cancer patients. In addition, we attempted to assess the clinicopathological implications for pancreatic cancer of the variant 2 (v2) isoform using a recently developed monoclonal antibody against a v2 epitope. The expression of CD44 variants was evaluated immunohistochemically in paraffin-embedded pancreatic cancer tissues from 42 patients who were confirmed surgically and histologically to have received curative resection. An indirect immunoperoxidase method was used with monoclonal antibodies against epitopes of the standard (CD44s) portion, v6 and v2. Protein expression data were evaluated statistically for any correlations with the length of survival or with histological parameters. The expression of CD44v6 and v2 was observed only in tumor cells, if at all. On the other hand, expression of total CD44 (including CD44v, as well as CD44s) was observed in both tumors and adjacent normal sites. Tumor tissue from 21 (50%) and 16 (38%) patients showed positive immunoreactivity with mAb 2F10 (anti-CD44v6) and mAb M23.6.1 (anti-CD44v2), respectively. The expression of CD44v6 and v2 was correlated with decreased overall survival (P = 0.0160 and P = 0.0125, respectively). A significant correlation was obtained between CD44v2 peptide expression and vessel invasion (P = 0.026). These results suggest that CD44v2 and CD44v6 may be useful markers for poor prognosis in curatively resected primary pancreatic cancer. PMID- 9849583 TI - Direct activation of human CD8+ cytotoxic T lymphocytes by interleukin-18. AB - Direct activation of human cytotoxic T lymphocytes (CTL) by interleukin (IL)-18 was observed in a system in which CTL effective against autologous tumor cells were generated. Peripheral blood mononuclear cells (PBMC) from tumor-bearing patients, after removal of natural killer (NK) cells, were cultured in a medium containing IL-1, -2, -4, and -6, with or without IL-18, and stimulated with autologous tumor cells. IL-18 increased the activity of the CTL and the proportion of autologous CD8+ T cells present after 28 days in the induction culture. When purified CD8+ T cells were cultured in the presence of IL-18 and IL 2 for 7 days, the CTL showed enhanced cytotoxic activity against autologous tumor cells. Moreover, a purified CD8+ T cell population, which did not exhibit any apparent cytotoxic activity against autologous tumor cells, displayed cytotoxic activity after 7-day incubation with IL-18. These results suggest that IL-18 may be useful to generate autologous CTL in humans and may thereby contribute to adoptive immunotherapy for tumors. PMID- 9849585 TI - In vivo efficacy and tumor-selective metabolism of amrubicin to its active metabolite. AB - The tissue distribution of a novel antitumor anthracycline antibiotic, amrubicin, was studied using seven human tumor xenografts implanted into nude mice, in order to identify the principal factors determining its therapeutic efficacy. We found a good correlation between the level of the metabolite amrubicinol in the tumor and the in vivo efficacy. High metabolic activity of amrubicin to amrubicinol was detected in tumor tissue homogenates, especially in cell lines highly sensitive to amrubicin in vivo. In contrast to amrubicin, the administration of amrubicinol showed less tumor-selective toxicity in these human tumor xenograft models. These data indicate that the tumor-selective metabolism of amrubicin to amrubicinol resulted in a tumor-selective disposition of amrubicinol, leading to good efficacy in in vivo experimental therapeutic models. PMID- 9849584 TI - Different effects of FK317 on multidrug-resistant tumor in vivo and in vitro. AB - FK317, a novel substituted dihydrobenzoxazine, was examined for antitumor effects on multidrug-resistant (MDR) tumor cells in vitro and in vivo. In nude mice, FK317 markedly inhibited the growth of s.c. implanted KB-V1 vinblastine (VLB) resistant human epidermal carcinoma KB cells, as well as the parent cells (KB-3 1). However, KB-V1 showed much greater resistance to FK317 than to VLB and adriamycin (ADM) in the in vitro study. This resistance was reversed by the addition of verapamil, whereby intracellular accumulation of FK317 in the KB-V1 cells was also decreased. After incubation of FK317 in human and mouse blood, it was shown to be rapidly metabolized to a monodeacetylated form, and slowly metabolized further to a dideacetylated form. With the removal of the acetyl groups from FK317, resistance indexes in KB-V1 and SBC-3/ADM, ADM-resistant human lung carcinoma, decreased. In addition, photolabeling of P-glycoprotein with [3H]azidopine in KB-V1 plasma membrane was completely inhibited by FK317, but not by the deacetylated metabolites. These results indicate that FK317 is metabolized to deacetylated forms, which do not bind to P-glycoprotein and are incorporated into MDR cells, causing cytotoxic effects. PMID- 9849586 TI - Tumor-selective distribution of an active metabolite of the 9-aminoanthracycline amrubicin. AB - It has been reported that the 9-aminoanthracycline amrubicin shows good efficacy in human tumor xenograft models. We studied the disposition and metabolism of amrubicin in mice, in comparison with those of doxorubicin. Amrubicinol, a 13 hydroxy metabolite of amrubicin, which is 10 to 100 times more cytotoxic than amrubicin, was detected as a major metabolite in blood and tissues, and aglycones of amrubicin were also detected. A pharmacokinetic study revealed that amrubicin had a smaller distribution volume and a shorter half-life than doxorubicin. In several normal tissues, the levels of amrubicin and amrubicinol were lower than those of doxorubicin. In contrast, the tumor levels of amrubicinol in the mice treated with amrubicin were higher than those of doxorubicin in the mice treated with that drug, in tumors that are sensitive to amrubicin. These results suggest that the potent therapeutic activity of amrubicin is caused by the selective distribution of its highly active metabolite, amrubicinol, in tumors. PMID- 9849587 TI - Cytotoxicity of amrubicin, a novel 9-aminoanthracycline, and its active metabolite amrubicinol on human tumor cells. AB - Amrubicin, a completely synthetic 9-aminoanthracycline derivative, was previously shown to have potent antitumor activities against various human tumor xenografts. In this study, the in vitro activities of amrubicin and its major metabolite, amrubicinol, were examined using 17 human tumor cell lines. Amrubicinol was 5 to 54 times more potent than amrubicin, and as potent as doxorubicin, in inhibiting the growth of the cells following 3-day continuous drug exposure. Amrubicinol closely resembled doxorubicin in its profile of activities on the 17 human tumor cell lines. Cells were incubated with the drugs for 1 h, and the intracellular drug concentration and cell growth inhibition after 3 days were determined. Amrubicinol attained similar intracellular concentrations at lower medium concentrations compared to amrubicin, and the intracellular concentration of amrubicinol necessary to produce 50% cell growth inhibition was 3 to 8 times lower than that of amrubicin in 4 cell lines tested. Amrubicinol has a higher activity level inside the cells than does amrubicin. When cells were incubated with amrubicin for 5 h, a substantial amount of amrubicinol, more than 9% of that of amrubicin, was found in cells in 4 of the 8 cell lines tested. Amrubicinol may contribute to the in vitro growth-inhibitory effect of amrubicin on these cells. The results suggest that amrubicinol plays an important role in the in vivo antitumor effect of amrubicin as an active metabolite. PMID- 9849588 TI - Telomerase activity correlates with growth of transplantable osteosarcomas in rats treated with cis-diammine dichloroplatinum or the angiogenesis inhibitor AGM 1470. AB - To determine the role of telomerase activity in the growth of tumors in rats undergoing chemotherapy, a comparison of the volumes of telomerase-positive transplantable osteosarcomas was made in rats treated with the antineoplastic agent cis-diammine dichloroplatinum (CDDP) or the angiogenesis inhibitor O (chloroacetylcarbamoyl)fumagillol (AGM-1470). Male F344 rats, 8 weeks old, received transplants of macroscopic lung metastatic nodules into the subcutaneous back space and treatment was started on day 14 thereafter. CDDP was injected i.v. at doses of 0, 0.625, 1.25 and 2.5 mg/kg body weight (b.w.) and AGM-1470 was administered at total doses of 0, 2.5, 5 and 10 mg/kg b.w. over 2 weeks by osmotic pumps, also implanted into the subcutaneous back space, but remote from the transplanted tumors. On day 28, all animals were killed for measurement of transplanted tumor size and determination of telomerase activities by telomeric repeat amplification protocol (TRAP) assay. The results showed telomerase activity to be highly correlated with the treated/non-treated (T/C) tumor size ratio (r = 0.96, P < 0.0001). In a second experiment, CDDP at 2.5 mg/kg b.w. and AGM-1470 at 10 mg/kg b.w., these being the most effective doses, were given as in the first experiment, and animals were serially killed on days 14, 21, 28, 35 and 42. Tumors in rats treated with CDDP and AGM-1470 showed 18.2% and 20.5% of the control telomerase activity on days 35 and 21, respectively, when tumor growth was inhibited. However, on day 42, the activities increased to 46.5% and 92.5%, this correlating with re-growth (r = 0.73, P < 0.0001). These results suggest that decline of telomerase activity may be involved in tumor growth retardation induced by chemotherapeutic agents. This possibility clearly warrants further mechanistic studies. PMID- 9849589 TI - Elevation of cytoplasmic cytochrome c in radiation-induced apoptosis. AB - The signal transduction pathway involved in radiation-induced apoptosis remains unclear, especially as regards the site at which the primary effect of radiation occurs. In this study, we demonstrate that cytochrome c may be released from mitochondria into the cytosol after irradiation, but that direct irradiation of isolated mitochondria induces no elevation of cytochrome c release. These observations suggest that cytochrome c and mitochondria may be involved in the radiation-induced apoptotic pathway, but mitochondria might not be a target of radiation, and that a signal transduction pathway from an unknown target might exist upstream of mitochondria during radiation-induced apoptosis. PMID- 9849590 TI - Pathological characteristics of gastric carcinomas in the very old. AB - A total of 69 gastric carcinomas of very old people (aged > or = 85) were collected and pathologically analyzed in comparison with those of young to middle aged (30-39) and elderly (65-69) people, with special attention to their phase. In the very old, almost all (34/35) carcinomas in the early phase belonged to well-differentiated categories. In the advanced phase, half of them (17/34) were classified into poorly differentiated categories when determined from the predominant pattern, but a well-differentiated pattern almost always coexisted in the superficial site. Thus, the gastric carcinomas in the very old may principally develop as well-differentiated carcinomas which then progress to poorly differentiated carcinomas with time, in contrast to those of the young to middle-aged, most of which emerged from the very early phase as poorly differentiated lesions. The gross features of the carcinomas were also in line with these histological observations. The carcinomas of the elderly showed distinct similarity to those of the very old. The results suggest that poorly differentiated carcinomas of the young to middle-aged and the old may be better classified and analyzed separately in view of the generally recognized etiological (e.g., specifically close causal relationship with environmental factors of the intestinal-type carcinoma of the old) and biological (e.g., practically no tendency for hematogenous metastasis of the diffuse-type carcinoma of the young to middle-aged) differences, although in the General Rules for Gastric Cancer Study of Japan, both are placed in the same category, por (por2). PMID- 9849591 TI - The United States-Japan Cooperative Cancer Research Program seminar on "Cell cycle control and cancer". PMID- 9849592 TI - [Current status and perspectives of conventional radiologic diagnosis of the gastrointestinal tract]. AB - In the last twenty years the basic gastro-intestinal diagnostic procedures have had to record increasing losses. This is why the study of these examinations had becoming more and more endangered. From our experience and from literature review the question arises whether now or in future we can manage without the basic gastrointestinal diagnostic procedures. The answer is that both for efficiency and for costs the continuation of these procedures does not appear acceptable. The preservation of the X-ray techniques of examination should be object of discussion and agreement between radiologists, clinicians, endoscopists, and economically experienced professionals in administration. In order to meet the needs for urgent economic measures in hospitals, the increased use of investigation with water-soluble contrast media seems unavoidable. Therefore the examination techniques should be adjusted and optimised as described. PMID- 9849593 TI - [First experiences with the trauma radiologic PACS project at the University of Innsbruck Department of Radiology]. AB - Radiography in an emergency setting is a demanding task, in technical as well as in logistical terms. Rapid acquisition of high quality radiographs under difficult conditions, for example severely injured patients with multi-organ trauma is important. All image data and radiological reports must be transmitted to the trauma surgeon as soon as possible. To meet these needs with the promising means of modern X-ray technology, we decided to implement a completely digital X ray division inside the new outpatient trauma department. In the setting of an outpatient emergency department digital radiography and PACS cannot be realised without problems. We present our first experience with the installation and 10 month filmless routine in the X-ray division for traumatology outpatients at the university of Innsbruck department of radiology. PMID- 9849594 TI - [Dosage monitoring as a quality control instrument in digital radiography]. AB - PURPOSE: Automatic image processing procedures ensure a constant image quality. Overexposed images constitute a danger zone on account of a subjectively better image appearance (improved signal-to-noise ratio). We have examined the software "Dose-Monitoring" both for the identification of false exposures and as a quality control tool. MATERIALS AND METHODS: Firstly the correlation between dose and present IgM (mean grey tone from the crude histogram of a 12-bit image) was examined by exposing phantoms. A further-reaching comparison of the present IgM values of some MTRAs with the department average (nominal IgM) should detect tendencies for over- and underexposure. Finally a dose reduction in 12 forms of X ray examination was evaluated with regard to IgM course controls (median IgM). RESULTS: A high correlation (0.987) between dose and mean grey tone (IgM). In some MRTAs tendency for over-/underexposure was detected. The dose reduction of defined types of X-ray examination lead to a trend to lower mean IgM values. DISCUSSION: Numerous factors have an influence on the IgM value thus making its evaluation difficult, even so it is a suitable parameter for uncovering tendencies to incorrect exposure and for identifying processes requiring optimization. CONCLUSIONS: The value of this tool lies in the quality assurance of dose fluctuations and a possible dose reduction. PMID- 9849595 TI - [Appearance of choroidal melanoma on high resolution MRI using 1.5T with a dedicated surface coil in 200 consecutive patients]. AB - PURPOSE: Choroidal melanomas usually present a characteristic appearance in MRI. Differing characteristics can cause problems in differential diagnosis between melanomas and other masses in the globe. The purpose of this study was to evaluate the appearance of choroidal melanomas with MRI in a large consecutive patient group. METHODS: In a prospective study, 200 patients with choroidal melanomas were investigated with MRI using a 1.5 T scanner and a 5 cm surface coil. Both quantitative and qualitative evaluation of the resulting images was performed. RESULTS: 78.5% of the melanomas presented with homogeneous signal intensities within the tumor due to a homogeneous pigmentation whereas 21.5% of the melanomas demonstrated a mixed pigmentation. Signal intensities of the homogeneous melanomas in the plain T1-WI were moderately or markedly hyperintense compared to the vitreous in 29.3% and moderately or markedly hypointense in the T2-WI in 37.1%. An accompanying retinal detachment was found in 65.5% and an extraocular growth in 7.0%. CONCLUSIONS: In 10% to 37% we observed the typical well known MR appearance, including homogeneous high signal in the T1-WI and low signals in the T2-WI. For further differentiation, morphological criteria (e.g. shape, size, and position) were used, which are also discussed. PMID- 9849596 TI - [Gastric pouch dilatation: complications after laparoscopic implantation of a silicone gastric band in pathologic obesity]. AB - In 20 patients with morbid obesity a laparoscopic silicon gastric banding was installed using a LAP-BAND. All patients were examined postoperatively with water soluble oral contrast material according to the usual protocol. 8 weeks after the operation a second control with thickened barium sulfate was added to measure and adjust the width of the silicon band. There were no early postoperative complications. But in the follow-up three patients presented with a pathologic gastric pouch-dilatation. This severe complication, which can have different etiologies, requires early detection and specific therapy. PMID- 9849597 TI - [Comparative magnetic resonance imaging of renal space-occupying lesions with a high and low field MRI system]. AB - PURPOSE: A prospective study of the diagnostic accuracy and image quality of two MRI systems in the detection of renal tumors was investigated. MATERIALS AND METHODS: 34 patients with the clinical suspicion of a space-occupying renal lesion were examined by MRI with a low field (0.2 Tesla magnet) and a high field (1.5 Tesla magnet) for comparison. An "informed" and a "blind" observer evaluated all of the MR images. In addition, the signal-to-noise and contrast-to-noise ratios were evaluated as second quality parameters. RESULTS: In 29 cases the results could be compared with a confirmed release diagnosis. Diagnostic accuracy was comparable with both systems (Sensitivity for both observer on LF apparatus: 83%, HF apparatus: 81%) although the signal-to-noise and contrast-to-noise ratios were significantly poorer at low field. CONCLUSIONS: The low field system in comparable to the high field MRI for detection and differentiation of renal space occupying lesions. PMID- 9849598 TI - [3D CT of fractures: comparison of volume and surface reconstruction]. AB - PURPOSE: The comparison of volume rendering (VR) and surface rendering (SSD) for demonstrating fractures from spiral-CT data. MATERIALS AND METHODS: Standardized VR and SSD projections were produced from 50 spiral CT scans of 50 consecutive patients with fractures. Appropriate multiplanar reformattings (MPR) were used as the standard. RESULTS: SSD provided sufficient information in 31/50 cases. Results of VR were not significantly different (33/50). VR was superior in demonstrating 6/7 craniofacial fractures and 3/3 calcaneal fractures. SSD was superior for visualizing 2/3 shoulder fractures and 2/2 elbow fractures. CONCLUSION: VR is a flexible technique for the depiction of fractures and the assessment of spongiosa and articular surfaces at one setting. SSD is superior in the delineation of small dislocated fragments. PMID- 9849599 TI - [Color doppler ultrasonography in peripheral artery occlusive disease: continuous application of a signal enhancer]. AB - PURPOSE: The effect of continuous infusion of a signal enhancer on the diagnostic efficacy of Doppler ultrasound in peripheral artery disease was evaluated. METHODS: 35 patients with peripheral artery disease were investigated by Doppler ultrasound before and during infusion of a signal enhancer (Levovist). Femoral and popliteal arteries were examined. Angiographic diagnoses included occlusions (24) and stenoses (22). They were compared to plain and enhanced Doppler findings. Artefacts and effects on signal intensity were evaluated. RESULTS: An increased Doppler signal was observed starting 1 min 35 s (mean) after begin of the infusion. It persisted for 11 min 46 s (mean). Signal enhanced studies provided less over- (0/1) and underestimation (3/5) of the findings and evaluation in the adductor channel was less compromised. Signal enhanced studies highlighted collaterals. CONCLUSION: Doppler ultrasound in patients with peripheral arterial disease is improved by continuous application of a signal enhancer. PMID- 9849600 TI - [Detection of angiogenesis-dependent parameters by functional MRI: correlation with histomorphology and evaluation of clinical relevance as prognostic factor using cervix carcinoma as an example]. AB - PURPOSE: Purpose of this study is to compare functional MRI parameters with histomorphological markers of tumor microvessel density (MVD) and permeability (vascular endothelial growth factor) and to determine the ultimate value of both approaches by correlation with disease outcome in patients with primary cancer of the uterine cervix. METHOD: Pharmacokinetic parameters were calculated from contrast-enhanced dynamic MR imaging series in 37 patients with biopsy-proven primary cervical cancer. On the operative whole mount specimens, histomorphological markers of tumor angiogenesis (MVD, VEGF) were compared with the MRI-derived parameters. For MRI and histomorphological data, Kaplan-Meier survival curves were calculated and compared using logrank statistics. RESULTS: Significant (p < 0.05-0.01) associations were found between MVD and dynamic MRI parameters. No significant relationships were observed between VEGF expression and dynamic MRI parameters. Disease outcome was better assessed with dynamic MRI parameters than with the histomorphological approach. CONCLUSIONS: It is concluded that 1) the pathophysiological basis for the amplitude A in dynamic MRI is MVD but not VEGF expression; and 2) a functional, dynamic MRI approach may be more suited to assess angiogenic activity in terms of patient survival than current histomorphological-based markers of tumor angiogenesis. PMID- 9849601 TI - [Reduction of radiation exposure for patient and investigator in interventional radiography]. AB - PURPOSE: In a retrospective analysis of vascular interventional procedures, relations between parameters of the examination and radiation exposure of patient and medical personnel are examined. MATERIAL AND METHOD: 1208 vascular interventional procedures are evaluated. Interventional procedures are divided into three groups: percutaneous transluminal angioplasty, implantation of a stent, thrombolysis. RESULTS: Mean values of the radiation dose of patient and radiology personnel are reported for these examinations. The mean value of the radiation dose of the physician was 7 microSv (maximum 24 microSv), that of the patient 1548 cGy.cm2 (maximum 8485 cGy.cm2). CONCLUSION: The quantity of X-rays to the patient may be lowered by using pulsed fluoroscopy and by reducing the number of radiographs. Reduction of the number of radiographs may be achieved by using the last-image hold and the road mapping mode. The operator's dose can be decreased by using additional radiation protection systems like a MAVIG-radiation protection wall. The radiation dose reduction was 61% for the physician and 17% for the patient. PMID- 9849602 TI - [Percutaneous abscess drainage in Crohn disease]. AB - PURPOSE: To evaluate the success of percutaneous, CT-guided abscess drainage (PAD) in patients with Crohn's disease. METHODS: Within the last 5 years 8 patients with Crohn's disease were treated by PAD for intra-abdominal abscesses. A fistula was determined to be the cause in 4 patients. The abscesses arose spontaneously in 7 patients while one patient had a postoperative abscess. We used single lumen 10 F- and double lumen 12 F- and 14 F-catheters for drainage (duration of drainage 8-20 days). RESULTS: In all cases the abscess was successfully drained by PAD. However, an operation-free interval of at least three months was achieved in only two patients. A healing of the fistula was not attained in any of the 4 patients with a proven fistula. No enterocutaneous fistulas arose within the course of PAD. CONCLUSIONS: PAD is also useful for patients with Crohn's disease since it improves the starting situation for the necessary operative interventions. In most cases (especially with enterogenic fistulas), however, a long-lasting therapeutic result cannot be expected. PMID- 9849603 TI - [A new MR-(and CT-) compatible bone biopsy system: first clinical results]. AB - PURPOSE: To describe the development and clinical evaluation of a new, hand powered or alternatively motor-driven, MRI and CT compatible percutaneous bone biopsy system. MATERIALS AND METHODS: A new coaxial drill system (Cook Europe A/S) was designed for percutaneous, MR-guided bone biopsies and powered either by hand or an optional motor (10-250 rotations/minute using 6 bar [88.2 PSI] compressed air). The system has been used in 23 patients. Fourteen procedures were performed in a 1.5 T MR scanner (Philips-Gyroscan ACS-NT) which has an attached C-arm (Philips-BV 212-Angio) in case fluoroscopy is required, and 9 procedures in a CT scanner (Siemens-Somatom Plus). RESULTS: Driven by hand or by the pneumatic motor unit, the system achieved safe and accurate MR-guided access to all of the lesions and was even able to penetrate osteosclerotic lesions. MR- or CT-guided percutaneous biopsy yielded a correct diagnosis in all but 5 cases. No procedural complications occurred. CONCLUSION: MR-guided percutaneous bone biopsy performed with the new coaxial drill system was found to be safe and reliable, and suitable for obtaining histological specimens from skeletal lesions even when covered with thick cortical or sclerotic bone. PMID- 9849604 TI - [Simple and effective rt-PA thrombolysis design]. AB - PURPOSE: Assessment of a novel, simple and practical thrombolysis scheme for the purposes of a central hospital. MATERIALS AND METHODS: 314 thromboembolic lower extremity occlusions in 260 consecutive patients were treated by thrombolysis as follows: 6 ml rt-PA/h for 30 min; 3 ml rt-PA/h for another 30 min; 1 ml rt-PA/h for 7 hours, and 0.4 ml rt-PA until vessel patency. Antegrade introduction of a 5 F straight end-hole catheter through a 6 F, constantly heparin-flushed sheath, to the level of occlusion was followed by subsequent advancement of the catheter tip according to progress of lysis at time intervals of 2, 4, and 8 hours. The regimen was continued overnight. RESULTS: 82% of arterial occlusions were completely recanalized at the end of thrombolysis. Then, angioplasty was performed if appropriate. Best results (100% recanalization rate) were achieved by treating occlusions of the superficial femoral artery, even in cases of involvement of the entire length of the vessel, and isolated popliteal occlusions. A recanalization rate of 70% was achieved by treating distal crural vessel occlusion. Bypass grafts were recanalized in 50%. COMPLICATIONS: 3 x major hematoma, 2 x transluminal perforation; 16 x minor hematoma from the puncture site at the sheath, 4 x erythema, obviously reaction to rt-PA (together n = 25; 8%). CONCLUSIONS: We empirically found and solidly evaluated an effective thrombolysis scheme. Our results demonstrate that major advances are possible even in this well-grounded field. PMID- 9849605 TI - [Novel endoluminal stents for the treatment of arterial fistulas and perforations]. AB - PURPOSE: To evaluate the use of polycarbonate-polyurethane (Corethane) stented grafts for the treatment of arteriovenous fistulas and arterial perforations. MATERIALS AND METHODS: 4 patients at the age of 34 to 81 underwent percutaneous insertion of corethane endoluminal grafts. Two patients had an arterial fistula- one iliaco-iliac, one brachio-brachial. Two patients underwent treatment to seal arterial perforations that occurred in the external iliac artery in one and in a femoral venous bypass in another. RESULTS: In all cases the communication or leakage was sealed by the endograft in place but had to be combined with a detachable balloon in om case. In three patients, a longer follow-up became available, one patient died three weeks after treatment from unrelated causes. While two stent grafts remained open for 15 and 24 months, one patient experienced stent graft occlusion three months after treatment. CONCLUSIONS: Corethane stent grafts are helpful to seal short-neck arteriovenous fistulas and arterial perforations even under emergency conditions. PMID- 9849606 TI - [Intra-arterial application of the ACE inhibitor ramipril using a microperforated catheter for the prevention of neointimal proliferation after angioplasty in an animal model]. AB - PURPOSE: To evaluate the safety and benefit of a high-dose local drug administration via a microporous balloon catheter to prevent neointimal formation after balloon angioplasty. MATERIALS AND METHODS: In New Zealand white rabbits (n = 29) neointima formation was induced by balloon denudation. Additionally, the animals were fed a 0.5% cholesterol diet for 6 weeks. Directly after the denudation, local application of 1.8 mg ramipril (n = 7) or saline (n = 7) via a microporous balloon catheter was performed. Other animals (n = 7) were treated with a systemic ramipril administration. One control group were exclusively fed with a cholesterol diet (n = 8). 6 weeks after intervention the animals were sacrificed and morphometry of the vessels was performed. RESULTS: Local administration of ramipril resulted in a non-significant reduction of 17%. The local administration mode was combined with a significant increase in neointima formation. Systemic ramipril administration resulted in a 66% reduction of plaque area. CONCLUSIONS: The benefit of the local ramipril administration was diminished by the inherent vessel trauma. Systemic ramipril administration resulted in a significant reduction of neointimal proliferation in New Zealand white rabbits. PMID- 9849608 TI - [Real-time MRI with radial k-radial scanning technique for control of angiographic interventions]. AB - PURPOSE: To test the feasibility of real-time MR controlled guidance of field inhomogeneity catheters in vitro and in vivo as a first step to MR-guided angiographic interventions. METHODS: Applying a combination of radial scanning with the sliding window reconstruction technique, a frame rate of 23 low resolution images per second was achieved. Field inhomogeneity catheters were steered through a flow phantom and into the renal arteries of a pig. RESULTS: It was possible to visualize flow or, respectively, vessels and to depict catheter movements. This enabled real-time MR-guidance of the catheter into the renal arteries of the flow phantom and into those of the pig. CONCLUSIONS: The new technique yields a sufficiently high temporal resolution for MR-guidance of catheters through vessels. PMID- 9849607 TI - [Stability of iodinated contrast media in UV-laser irradiation and toxicity of the photoproducts]. AB - PURPOSE: In XeCl-Excimer laser angioplasty, unintended and possibly harmful interaction of the UV-laser light and the contrast media may occur due to the high concentration of contrast medium proximal to the occlusion or subtotal stenosis. METHODS: One ml of three nonionic monomeric contrast agents (iopromide, iomeprol, iopamidol), one nonionic dimeric (jotrolane), and one ionic monomeric (amidotrizoate) X-ray contrast agent were irradiated with a XeCl excimer laser (lambda = 308 nm, pulse duration 120 ns, 50 Hz) using a 9 French multifiber catheter (12 sectors). Up to 20,000 pulses (106 J) were applied. Using high performance liquid chromatography the amount of liberated iodide as well as the fraction of unchanged contrast media were measured. Cytotoxicity of the photoproducts was tested in a colony formation assay of human skin fibroblasts. The contrast agents were irradiated with 2000 pulses/ml (5.3 mJ/pulse; 10.6 J) and then added to the cell cultures for a period of three hours in a concentration of 10%. RESULTS: Excimer laser irradiation induced iodide liberation of up to 3.3 mg iodide/ml. Up to 19% of the contrast agents changed their original molecular structure. Incubation of irradiated contrast agents resulted in a significantly decreased potential for colony formation (p values ranging from 0.0044 to 0.0102) with significantly higher toxicity of amidotrizoate and iomeprol in comparison to iopromide, iotrolan, and iopamidol. DISCUSSION: Due to the cytotoxic photoproducts and the high level of liberated iodide, it is recommended to flush the artery with physiological saline solution before applying a pulsed excimer laser in human arterial obstructions in order to reduce the contrast agent concentration at the site of irradiation. PMID- 9849609 TI - [Magnetic resonance tomography in neurocutaneous melanosis]. PMID- 9849610 TI - [Chronic recurrent multifocal osteomyelitis of the lower jaw]. PMID- 9849611 TI - ["Pulmonary artery sling": a rare congenital vascular anomaly with tracheal stenosis. Value of diagnostic imaging]. PMID- 9849612 TI - [Localized fibrotic tumor of the pleura: imaging by MR tomography]. PMID- 9849613 TI - [Crohn disease of the esophagus with esophageal-pulmonary fistula]. PMID- 9849614 TI - Hammerhead ribozyme design and application. AB - The emerging knowledge about RNA-based enzymes has already had great impact on our concept of evolutionary history, making the 'RNA world' more likely. It may well have an equally important impact on the diagnostic and therapeutic practices of human and veterinary medicine in the next decade. We are not quite there yet. This review addresses the design and application of hammerhead ribozymes, two aspects of a conserved and most commonly studied and used enzymatically active entity among the RNA enzymes. The emerging picture is one of great diversity. There is at this stage no general cell model nor a clearly preferable ribozyme structure. Each and every cell line (and tissue) may be unique in that they vary with respect to structural requirements for optimal uptake, activity and stability of ribozymes. We may have seen only the tip of the iceberg when it comes to RNA-based enzymes and their roles in biology and medicine. PMID- 9849615 TI - The structure of human interferon-beta: implications for activity. AB - Interferons (IFNs) are potent extracellular protein mediators of host defence and homoeostasis. This article reviews the structure of human IFN-beta (HuIFN-beta), in particular in relation to its activity. The recently determined crystal structure of HuIFN-beta provides a framework for understanding of the mechanism of differentiation of type I IFNs by their common receptor. Insights are generated by comparison with the structures of other type I IFNs and from the interpretation of existing mutagenesis data. The details of the observed carbohydrate structure, together with biochemical data, implicate the glycosylation of HuIFN-beta, which is uncommon among type I IFNs, as an important factor in the solubility, stability and, consequently, activity of the protein. Finally, these structural implications are discussed in the context of the clinical use of HuIFN-beta. PMID- 9849616 TI - Chimeras of human lysozyme and alpha-lactalbumin: an interesting tool for studying partially folded states during protein folding. AB - Protein folding is an extremely active field of research where biology, chemistry, computer science and physics meet. Although the study of protein folding intermediates in general and equilibrium intermediates in particular has grown considerably in recent years, many questions regarding the conformational state and the structural features of the various partially folded intermediate states remain unanswered. Performing kinetic measurements on proteins that have had their structures modified by site-directed mutagenesis, the so-called protein engineering method, is an obvious way to gain fine structural information. In the present review, this method has been applied to a variety of proteins belonging to the lysozyme/alpha-lactalbumin family. Besides recombinants obtained by point mutations of individual critical residues, chimeric proteins in which whole structural elements (10-25 residues) from alpha-lactalbumin were inserted into a human lysozyme matrix are examined. The conformational properties of the equilibrium intermediate states are discussed together with the structural characterization of the partially unfolded states encountered in the kinetic folding pathway. PMID- 9849617 TI - The effects of environmental hormones on reproduction. AB - Considerable attention has been given in the past few years to the possibility that man-made chemicals (xenobiotics) in the environment may pose a hazard to human reproductive health. The endocrine-disrupting effects of many xenobiotics can be interpreted as interference with the normal regulation of reproductive processes by steroid hormones. Evidence reviewed here indicates that xenobiotics bind to androgen and oestrogen receptors in target tissues, and to androgen binding protein and to sex hormone-binding globulin. Although environmental chemicals have weak hormonal activity, their ability to interact with more than one steroid-sensitive pathway provides a mechanism by which their hazardous nature can be augmented. A given toxicant may be present in low concentration in the environment and, therefore, harmless. However, we are not exposed to one toxicant at a time, but, rather, to all of the xenobiotics present in the environment. Therefore, numerous potential agonists/antagonists working together through several steroid-dependent signalling pathways could prove to be hazardous to human reproductive health. PMID- 9849618 TI - CD100 is a leukocyte semaphorin. AB - CD100 was originally described as an activation molecule on the surface of human T lymphocytes. Its triggering through distinct epitopes leads to different signals of costimulation with phorbol myristate acetate (PMA) or with CD3 and CD2. Interestingly, CD100 was shown to associate with different partner molecules in T cells. First, CD100 can associate with CD45, a key molecule with protein tyrosine phosphatase activity involved in T-cell transduction: this association is physical and has functional consequences for both partners. Second, CD100 interacts in its cytoplasmic domain with a Ser/Thr kinase for which it represents a preferential substrate. Recently, CD100 was identified as a member of the semaphorin gene family. This family comprises approximately 20 structurally related proteins. The first semaphorins were identified in the developing nervous system. Function has been shown for only some of them and involves repulsion during growth cone guidance. Since CD100 was the first semaphorin identified in the immune system, this raises the possibility of the involvement of members of the semaphorin family in other physiological phenomena outside the nervous system. PMID- 9849619 TI - Oxygen consumption and biosynthetic function in perfused liver from rats at different stages of development. AB - Changes in mitochondrial function were studied in perfused liver from rats aged 24-365 days. Oxygen consumption together with the rates of gluconeogenesis, urea synthesis and ketogenesis were determined. Basal mitochondrial respiration as well as the ability of the liver to synthesize glucose, urea and ketone bodies declined from 24- to 365-day-old rats. On the other hand, on transition from 24 to 60 days the liver oxidation rate of hexanoate, sorbitol and glycerol is enhanced, but not of ketone bodies or palmitate. Our results show that the transition from weaning to middle age is accompanied by defined changes in hepatic substrate oxidation. From the observed time course of the decrease in basal and substrate-stimulated oxygen consumption, it is concluded that in rat liver cells a decline in respiratory chain function, long-chain fatty acid and ketone body metabolism, gluconeogenesis and ureogenesis occurs at a relatively early life stage. PMID- 9849620 TI - Characterization of a hypermutable strain of Drosophila simulans. AB - A hypermutable strain of Drosophila simulans that originated from a single spontaneous mutant male was characterized. Seven different mutations were isolated from roughly 100 generations of offspring. The genetic analysis of the viable mutants showed two mutations on the X chromosome, one in the lozenge locus and the other in the ruby gene. The autosomic mutations characterized were a dpp heldout-like, a blistered-like and a homoeotic dominant mutant with an antenna-to leg transformation and ectopic eyes that we called Zoinho-napata. PMID- 9849621 TI - Molecular basis of film formation from a soybean protein: comparison between the conformation of glycinin in aqueous solution and in films. AB - Fourier transform infrared spectroscopy has been used to investigate the conformational changes of glycinin. a major storage protein of soybean seeds, upon film-forming. The results show that the secondary structure of glycinin is mainly composed of a beta-sheet (48%) and unordered (49%) structures. The amide I band of glycinin in film-forming conditions, i.e. in alkaline media and in the presence of plasticizing agent, reveals the conversion of 18% of the secondary structure of the protein from the beta-sheet (6%) and random coil (12%) to the alpha-helical conformation due to the helicogenic effect of the ethylene glycol used as the plasticizing agent. Conformational changes also occur upon the film forming process leading to the formation of intermolecular hydrogen-bonded beta sheet structures. Results obtained from other plant families indicate that, whatever the origin and conformation of protein, formation of films leads to the appearance of intermolecular hydrogen-bonded beta-sheet structures, suggesting that this type of structure might be essential for the network formation in films. Thus, it is hypothesized that, in the film state, intermolecular hydrogen bonding between segments of beta-sheet may act as junction zones in the film network. This study reveals for the first time that there is a close relationship between the conformation of proteins and the mechanical properties of films. PMID- 9849622 TI - Effect of local sequence inversions on the crystalline antiparallel beta-sheet lamellar structures of periodic polypeptides: implications for chain-folding. AB - The crystal structure and texture of the monodisperse periodic polypeptide [(AG)3EG(GA)3EG]10 (poly(+/-AG)3EG: A=alanine, G=glycine, E=glutamic acid) were analyzed by X-ray diffraction, Fourier transform infrared spectroscopy, and electron microscopy. Structure determination was aided by comparison with the recently described structure for the related periodic polypeptide [(AG)3EG]36 by Krejchi et al. (Macromolecules 1997;30:5012). Texture-oriented samples of poly(+/ AG)3EG were obtained by crystallization of the polymer from aqueous formic acid solution. The evidence supports an antiparallel (ap) beta-sheet protein structure and the X-ray diffraction signals index on an orthorhombic unit cell with parameters: a=0.950 nm (hydrogen-bond direction), b=1.052 nm (apbeta-sheet stacking direction), c=6.95 nm (chain direction). The absence of the (010) diffraction signal, a prominent signal in the poly(AG)3EG diffraction pattern, implies that the apbeta-sheets are 'apolar', i.e. both surfaces are equally populated with alanyl methyl groups. Selective line broadening of wide-angle diffraction signals with l not equal to 0 gives an estimated crystal size of approximately/= 4 nm in the chain direction. This observation, coupled with the appearance of low-angle particle interference peaks, indicates a crystal thickness considerably less than the chain length and suggests an adjacent-re entry chain-folded lamellar structure incorporating the apbeta-sheet architecture. The polypeptide folds through gamma-turns, in-phase with the pseudo octapeptide repeat; the glutamic acid residues occur on the lamellar surfaces. These results and those from the crystalline lamellae of poly(AG)3EG suggest that beta-turns are not compatible with these repetitively stacked apbeta-sheet structures. This implies that intersheet interactions of alanyl methyl groups and glycyl alpha-protons are not sufficiently strong to dictate the folding geometry in these structures. PMID- 9849623 TI - Chitin-graft-poly(2-methyl-2-oxazoline) enhanced solubility and activity of catalase in organic solvent. AB - Catalase from bovine liver was lyophilized from an aqueous solution containing chitin-graft-poly(2-methyl-2-oxazoline) (3), which was synthesized by the reaction of 52% deacetylated chitin (1) with living poly(2-methyl-2-oxazoline) (2). The rate of consumption of H2O2 in chloroform catalyzed by the lyophilized catalase with 3 was enhanced more than 10 times that by catalase without 3. The dispersibility and solubility of lyophilized catalase with 3 in chloroform were improved in comparison with catalase itself. PMID- 9849624 TI - Xanthan-galactomannan interactions as related to xanthan conformations. AB - The influence of xanthan conformation on the physicochemical behaviour of their mixtures with galactomannan from Schizolobium parahybae mannose:galactose ratio (M/G=3), was studied by viscoelastic measurements, differential scanning calorimetry (DSC) and chiroptical (circular dichroism) methods. The results suggested a more effective interaction of the galactomannan with disordered xanthan segments, which are more abundant in low salt concentrations but are still present in lower proportion at temperatures lower than the temperature of xanthan conformational transition (Tm). The dependence of ellipticity with temperature in a circular dichroism (CD) spectra suggested an ordering of the xanthan chains induced by galactomannan at the temperature of gel formation (Tg approximately 25 degrees C), under conditions where xanthan alone exhibits a disordered conformation. The lower Tg value found (approximately 25 degrees C) compared with that (60 degrees C) usually described in the literature is certainly related to the M/G ratio and the galactosyl unit distribution along the mannan main chain. PMID- 9849625 TI - Circular dichroism study of ribonuclease A mutants containing the minimal structural requirements for dimerization and swapping. AB - Four residues Pro19. Leu28, Cys31 and Cys32 proved to be the minimal structural requirements in determining the dimeric structure and the N-terminal segment swapping of bovine seminal ribonuclease, BS-RNase. We analyzed the content of secondary and tertiary structures in RNase A, P-RNase A, PL-RNase A, MCAM-PLCC RNase A and MCAM-BS-RNase, performing near and far-UV CD spectra. It results that the five proteins have very similar native conformations. Thermal denaturation at pH 5.0 of the proteins. studied by means of CD measurements. proved reversible and well represented by the two-state N<==>D transition model. Thermodynamic data are discussed in the light of the structural information available for RNase A and BS-RNase. PMID- 9849626 TI - Characterisation of the polysaccharide produced by Acetobacter xylinum strain CR1/4 by light scattering and atomic force microscopy. AB - The molecular weight of the extracellular polysaccharide (CR1/4) produced by Acetobacter xylinum strain CR1/4 has been shown to be dependent upon growth conditions. Under normal growth conditions a high molecular weight polysaccharide ( > 1 x 10(6) Da) is produced. Maintaining the pH at 5 results in an order of magnitude increase in the total yield of polysaccharide, but also an order of magnitude decrease in molecular weight. Analysis of the CR1/4 polysaccharides by the techniques of atomic force microscopy and static light scattering suggests that they are double helices. In solution the molecules behave as stiff coils with a Kuhn statistical segment length of 325 nm. PMID- 9849627 TI - Architecture of the sugar binding sites in carbohydrate binding proteins--a computer modeling study. AB - Different sugars, Gal, GalNAc and Man were docked at the monosaccharide binding sites of Erythrina corallodenron (EcorL), peanut lectin (PNA), Lathyrus ochrus (LOLI), and pea lectin (PSL). To study the lectin-carbohydrate interactions, in the complexes, the hydroxymethyl group in Man and Gal favors, gg and gt conformations respectively, and is the dominant recognition determination. The monosaccharide binding site in lectins that are specific to Gal/GalNAc is wider due to the additional amino acid residues in loop D as compared to that in lectins specific to Man/Glc, and affects the hydrogen bonds of the sugar involving residues from loop D, but not its orientation in the binding site. The invariant amino acid residues Asp from loop A, and Asn and an aromatic residue (Phe or Tyr) in loop C provides the basic architecture to recognize the common features in C4 epimers. The invariant Gly in loop B together with one or two residues in the variable region of loop D/A holds the sugar tightly at both ends. Loss of any one of these hydrogen bonds leads to weak interaction. While the subtle variations in the sequence and conformation of peptide fragment that resulted due to the size and location of gaps present in amino acid sequence in the neighborhood of the sugar binding site of loop D/A seems to discriminate the binding of sugars which differ at C4 atom (galacto and gluco configurations). The variations at loop B are important in discriminating Gal and GalNAc binding. The present study thus provides a structural basis for the observed specificities of legume lectins which uses the same four invariant residues for binding. These studies also bring out the information that is important for the design/engineering of proteins with the desired carbohydrate specificity. PMID- 9849628 TI - Plants used in Guatemala for the treatment of protozoal infections. I. Screening of activity to bacteria, fungi and American trypanosomes of 13 native plants. AB - Extracts were prepared from 13 native plants used for the treatment of protozoal infections. Activity against bacteria and fungi was demonstrated by dilution procedures; Trypanosoma cruzi was evaluated in vitro against epimastigote and trypomastigotes and in vivo against trypomastigotes. In active extracts, toxicity was evaluated by Artemia salina nauplii, oral acute toxicity (1-5 g/kg) and oral and intraperitoneal subacute toxicity in mice (500 mg/kg). From the plants screened, six showed activity (< or = 2 mg/ml) against bacteria, three against yeasts, five against Microsporum gypseum and five against T. cruzi in vitro and/or in vivo. In vitro and in vivo activity was demonstrated by Neurolaena lobata and Solanum americanum; in vitro or in vivo activity was shown by Acalypha guatemalensis, Petiveria alliacea and Tridax procumbens. Toxicity studies showed that extracts from S. americanum are toxic to A. salina (aqueous, 160 ppm). None showed acute or oral toxicity to mice; S. americanum showed intraperitoneal subacute toxicity. PMID- 9849629 TI - Uterotonic properties of the methanol extract of Monechma ciliatum. AB - The oxytocic activity of the hot methanol extract (HME) of the leaves of Monechma ciliatum was compared with other uterine stimulants like ergometrine, oxytocin, 5 hydroxytryptamine (5-HT), acetylcholine (ACh) and prostaglandins (PGs) E2 and F2alpha (PGE2 and PGF2alpha) in the presence of some antagonists in an attempt to explain the mechanism of action of the extract. The effects of the reference drugs on uteri isolated from rats pretreated with HME for 2 weeks were also observed. Atropine blocked the effect of ACh and partially blocked those of HME while L-366-948 blocked only the effect of oxytocin. Indomethacin inhibited the effects of HME as well as all the other drugs, except the PGs and ACh. D-600 blocked the effect of all the drugs including HME. Methysergide antagonised only the effect of 5-HT and partially blocked ergometrine. Prolonged treatment altered the uterine musculature and the activity profile of the reference drugs. These results suggest that the HME may be acting by more than one mechanism to contract the uterus and explains the mechanism of the anti-implantation activity of the plant. PMID- 9849630 TI - Suppressive effect of cyclophosphamide-induced toxicity by Withania somnifera extract in mice. AB - Administration of Withania somnifera extract (Solanaceae) was found to significantly reduce leucopenia induced by cyclophosphamide (CTX) treatment. The total WBC count on the 12th day of the CTX-treated group was 3720 cells/mm3 and that of CTX along with Withania was 6120 cells/mm3. Treatment of Withania along with CTX was found to significantly (P < 0.001) increase the bone marrow cellularity (13.1 x 10(6) cells/femur) compared to CTX alone treated group (8 x 10(6) cells/femur). Administration of Withania extract increased the number of alpha-esterase positive cells (1130/4000 cells) in the bone marrow of CTX treated animals, compared to the CTX-alone treated group (687/4000 cells). The major activity of Withania somnifera may be the stimulation of stem cell proliferation. These studies indicate that Withania somnifera could reduce the cyclophosphamide induced toxicity and its usefulness in cancer therapy. PMID- 9849631 TI - Modulation of American ginseng on brainstem GABAergic effects in rats. AB - Single neurons in the region of the medial nucleus tractus solitarius (NTS), responding or not responding to gastric vagal branch stimulation, were recorded in an in vitro neonatal rat brainstem-gastric preparation. The spontaneous activity of the majority of these two types of NTS units was inhibited by GABA(A) receptor agonist, muscimol (30 microM), and this inhibition (approximately 52% compared to 100% of the control level) could be antagonized by selective GABA(A) receptor antagonist, bicuculline (10 microM). Application of Panax quinquefolium L. extracts (3.0 microg/ml) into the brainstem compartment of the preparation also significantly reduced the discharge rate of these NTS neurons (approximately 27% compared to the control level), but this reduction could not be reversed by bicuculline (10 microM). Pretreatment with Panax quinquefolium L. (3.0 microg/ml) significantly decreased the NTS inhibitory effects induced by muscimol (30 microM), approximately from 51 to 33%. Our results demonstrated the interactions of Panax quinquefolium L. with ligand-bindings of GABA(A) receptors, and the modulation of the brainstem GABAergic mechanism by Panax quinquefolium L. Our data suggest that the regulation of GABAergic neurotransmission may be an important action of Panax quinquefolium L. PMID- 9849632 TI - Isolation and identification of active compounds from Drimys winteri barks. AB - The barks of Drimys winteri are used in folk medicine as a remedy to treat several diseases, including dolorous processes. Previous pre-clinical experiments carried out in our laboratories revealed that the hydroalcoholic extract of this plant showed anti-allergenic, anti-inflammatory and antinociceptive properties. Such promising results led us to determine the analgesic compounds present in D. winteri. Through conventional chromatographic procedures with fractions of CH2Cl2 and EtOAc obtained from methanolic extract, it was found that polygodial (1), 1 beta-(p-methoxycynnamyl) polygodial (2), taxifolin (3) and astilbin (4), are the main components of these fractions. Compounds 1 and 2 exhibited marked antinociceptive action by intraperitoneal and oral routes against acetic acid induced abdominal constrictions in mice, suggesting that they are responsible, at least partially, for the antinociceptive effects of this plant. In addition, both compounds were notably more potent than aspirin and acetaminophen, two well-known drugs used here as comparison. PMID- 9849633 TI - Neuropharmacological activities of the crude alkaloidal fraction from stems of Tabernaemontana pandacaqui Poir. AB - The crude alkaloidal (CA) fraction from the stem of Tabernaemontana pandacaqui Poir. was studied for its pharmacological activity on the central nervous system (CNS) of animals. The CA fraction was found to produce symptoms of CNS depression in conscious rats and mice, viz. reduction in spontaneous motility, potentiation of pentobarbital sleeping time, prolongation of latency of convulsions induced by pentylenetetrazole and antinociception. However, the fraction could not antagonize oxotremorine-induced tremor. The observations suggest that the CA fraction possesses a CNS depressant activity. PMID- 9849634 TI - Subchronic toxicity study in mice fed Spirulina maxima. AB - The purpose of this study was to evaluate the toxicity of Spirulina maxima, a blue-green alga used as food supplement and food coloring, after 13 weeks of treatment. Groups of ten mice of each sex were given S. maxima in the diet at concentrations of 0 (control), 10, 20 or 30% (w/w) for 13 weeks. The alga ingestion had no effect on behavior, food and water intake, growth or survival. Terminal values in hematology and clinical chemistry did not reveal differences between treated and control groups. However, male and female mice showed significant changes in serum cholesterol levels at 20 and 30% algal concentrations, but a toxic effect of S. maxima was excluded. Post-mortem examination revealed no differences in gross or microscopic findings. Our results show that S. maxima up to high feeding levels did not produce adverse effects in mice after subchronic treatment. PMID- 9849635 TI - Preliminary antifungal and cytotoxicity studies of extracts of Ritchiea capparoides var. longipedicellata. AB - Crude extracts obtained from the leaves, stem bark and roots of Ritchiea capparoides var. longipedicellata were screened for in vitro antifungal activity using the agar tube dilution method. The leaf hexane, leaf methanol, stem bark methanol and root methanol extracts were tested using ten clinical strains of fungi at a concentration of 200 and 400 microg/ml, respectively. At 400 microg/ml, all four extracts inhibited the growth of six of the ten test fungi used in the study. Inhibition of the growth of Aspergillus niger by the extracts was also seen but the activity was low and the leaf hexane and root methanol extracts inhibited the growth of Drechslera rostrata. Only the leaf hexane extract was active against Curvularia lunata, while the growth of Candida albicans was not inhibited by any of the extracts. The inhibition of growth of almost all the microorganisms decreased at 200 microg, griseofulvin was included as a reference compound and methanol as the control. Preliminary cytotoxicity tests were done with the four extracts using the larvae of the brine shrimp, Artemia saline. The extracts were however found to be relatively non-toxic as each extract had an LD50 value greater than 1000 microg/ml. PMID- 9849636 TI - Evaluation of the hypoglycaemic effect of Memecylon umbellatum in normal and alloxan diabetic mice. AB - Oral administration of an alcoholic extract of the leaves of Memecylon umbellatum led to a significant lowering of serum glucose levels in normal and alloxan induced diabetic mice. Although the mechanism of action is unknown, the magnitude of glucose reduction in both the animal models is encouraging. The results indicate that M. umbellatum has interesting possibilities as a source of oral hypoglycaemic agents. PMID- 9849637 TI - Antimicrobial study of the resinous exudate and of diterpenoids isolated from Eupatorium salvia (Asteraceae). AB - The antimicrobial properties of the resinous exudate from twigs and leaves of Eupatorium salvia were tested against five gram-negative and five gram-positive bacteria. Comparison of the antimicrobial activities of 7-hydroxy-8(17)-labden-15 oic acid (salvic acid) and of its acetate, both compounds isolated from the plant, with that of the crude extract suggested that the latter ester derivative was the major active component in the exudate. These results validate the vernacular medicinal use of Eupatorium salvia as an antiseptic agent. PMID- 9849638 TI - The preliminary isolation of several antibacterial compounds from Combretum erythrophyllum (Combretaceae). AB - Freeze dried ground leaves of Combretum erythrophyllum were extracted with different extractants to determine if they contain inhibitors of pathogenic bacteria. Acetone yielded the most compounds inhibiting the growth of Staphylococcus aureus using bioautography of thin layer chromatography plates. Acetone also extracted many different compounds and group separation was attempted by solid phase extraction on reverse and normal phase silica gel. Both techniques separated the bioactive components to a degree. The best group separation results, however, were obtained using liquid/liquid extraction. The six fractions obtained inhibited the four test organisms to different degrees. S. aureus was the most sensitive (100%) followed by Enterococcus faecalis (36%), Escherichia coli (11%) and Pseudomonas aeruginosa (3%). With S. aureus as test organism, the chloroform soluble fraction contained by far the largest quantity of inhibiting components (100%), followed by the fractions soluble in water (23%), 35% methanol in water (18%), butanol (5%), carbon tetrachloride (2%) and hexane (traces). The lowest minimum inhibitory concentration for S. aureus was 0.05 mg/ml at this stage of purification compared to MIC values of 0.08 and 0.16 mg/ml for ampicillin and chloramphenicol. There were at least 14 different inhibitors with a wide range of polarities present in the different fractions. The polar components apparently did not contain polysaccharides and were probably basic according to the chromatographic behaviour. PMID- 9849639 TI - Identification of cytochrome P450 isozymes involved in metabolism of the alpha1 adrenoceptor blocker tamsulosin in human liver microsomes. AB - 1. The in vitro human liver metabolism of the alpha1-adrenoceptor blocker tamsulosin was investigated. When 14C-tamsulosin was incubated with human liver microsomes, it was converted to five known urinary metabolites and at least three unknown metabolites. Of the former group, the predominant metabolite was the O deethylated metabolite (M-1), followed by the o-ethoxyphenoxy acetic acid (AM-1) and the m-hydroxylated metabolite (M-3). 2. There was a good linear relationship between AM-1 formation and testosterone 6beta-hydroxylase activity in microsomes from each of 10 individual donors. The rate of M-1 formation also correlated with the same activity, albeit the correlation curve did not pass through the origin. By contrast, the rates of M-3 and the O-demethylated metabolite (M-4) formation correlated with dextromethorphan O-demethylase activity. 3. Ketoconazole strongly inhibited AM-1 formation and reduced that of M-1 by c. 60%. Immunoinhibition studies using anti-rat antibodies supported these results. The formation of M-3 and M-4 was inhibited by quinidine and sparteine. 4. It is concluded that formation of tamsulosin metabolites, AM-1 and M-1, is catalysed by CYP3A4 whereas that of M-3 and M-4 is catalysed by CYP2D6. However, minor contributions from other CYPs cannot be excluded. PMID- 9849640 TI - Xenobiotic-metabolizing enzymes in pig nasal and hepatic tissues. AB - 1. A study of xenobiotic-metabolizing enzyme activity of the olfactory and respiratory epithelium in the pig was undertaken. The results indicated that porcine olfactory mucosa contains all the components of the P450 system. 2. Monooxygenase activities were much higher in olfactory than in respiratory microsomes, and the olfactory activities dependent on CYP2A were higher than those in the liver. By contrast, the olfactory monooxygenases associated with CYP2E1 were poorly or not detected, whereas CYP2G1 and a protein immunorelated to CYP1A2 were expressed in the olfactory epithelium. 3. The activities of several non oxidative enzymes (glutathione S-transferase, UDP-glucuronyl transferase, epoxide hydrolase, DT-diaphorase, benzaldehyde and propionaldehyde dehydrogenases, and various esterases) were also determined in porcine tissues and were found to be higher in the olfactory than in the respiratory mucosa, but lower or similar to those in liver. 4. An unexpected finding was a higher activity of olfactory UDP-GT compared with that of liver when 1-naphtol but not p hydroxybiphenyl (a good substrate for a specific olfactory UDP-GT(olf) in bovine and rat) was used as substrate, suggesting a porcine specific expression of UDP GT isoforms. 5. The results taken together indicate that the olfactory epithelium of mammals has a similar cytochrome P450 profile with the CYP2A and CYP2G1 as dominant isoforms, whereas the olfactory non-oxidative enzymes appear qualitatively and quantitatively expressed to different extents. PMID- 9849641 TI - Metabolic activity of fresh and cryopreserved dog hepatocyte suspensions. AB - 1. Dog hepatocytes were cryopreserved at 6 x 10(6) viable cells/ml in a suspension buffer containing 10% DMSO and were stored in liquid nitrogen. 2. The exclusion of trypan blue dye was 96 +/- 2 and 85 +/- 9% in fresh and cryopreserved (CP) hepatocytes, respectively. Albumin synthesis was unaffected by freezing. 3. Ethoxycoumarin and ethoxyresorufin O-deethylase activities were equivalent in fresh and CP hepatocytes. 4. The profile of testosterone metabolism was unaffected by freezing. Total hydroxylase activities were 815 +/- 33 pmol/min/10(6) cells in freshly isolated whole hepatocytes and 463 +/- 24 pmol/min/10(6) CP whole hepatocytes, but they were equivalent in fresh and CP hepatocyte homogenates supplemented with 250 microM NADPH. 5. Phase 2 enzymes were functional in freshly thawed CP hepatocytes but they required exogenous addition of cofactors (20 microM UDPGA and 1.7 microM PAPS). 6. When placed in suspension for longer times, fresh and CP cell viabilities were 88 +/- 6 and 64 +/- 2% after 4 h. ECOD and EROD activities were equivalent in fresh and CP hepatocyte suspensions, over 4 h. Testosterone hydroxylase activities were well maintained in fresh cell suspensions but they declined to 63 +/- 6% of the initial activity after 4 h in CP hepatocytes. 7. These results indicate that CP dog hepatocytes are a suitable in vitro system for xenobiotic metabolism since enzyme functions in CP hepatocytes were stabilized. Cofactors in freshly thawed CP hepatocytes should be measured and controlled for optimal use. PMID- 9849642 TI - Drug-metabolizing enzyme induction by 2,2'-dipyridyl, 1,7-phenanthroline, 7,8 benzoquinoline and oltipraz in mouse. AB - 1. Changes in the major hepatic drug-metabolizing enzymes by compounds identified as atypical inducers (multienzyme response but devoid of cytochrome P450-inducing ability) in rat were investigated in mouse. Animals were treated with 1,7 phenanthroline, 2,2'-dipyridyl, 7,8-benzoquinoline and oltipraz at 75 and 150 mg/kg daily for 3 days. 2. UDP-glucuronosyltransferase (UGT) activities showed only limited changes, UGT activity towards 4-nitrophenol and 1-naphthol was induced by the 75 mg/kg dose of 2,2'-dipyridyl and UGT activity towards morphine was induced by 150 mg/kg doses of 7,8-benzoquinoline and oltipraz. UGT activity towards oestrone was not induced by any treatment regimen and showed a decrease following treatment with the lower dose of 7,8-benzoquinoline. 3. In contrast with the limited effect on UGT activities, glutathione S-transferase and NAD(P)H:quinone oxidoreductase activities were significantly elevated by most compounds. Glutathione S-transferase activity was significantly elevated by the 150 mg/kg dose of 1,7-phenanthroline (73%), 2,2'-dipyridyl (52%) and oltipraz (75%), and also the lower dose of 1,7-phenanthroline (47%). NAD(P)H:quinone oxidoreductase activity was significantly elevated by the higher dose of all N heterocycles (155-323%) as well as the lower dose of 1,7-phenanthroline (180%). 4. In contrast with the effect previously seen in rat, 7,8-benzoquinoline significantly elevated mouse cytochrome P450 concentration but not 7 ethoxyresorufin O-dealkylase activity. As in rat, no N-heterocycle-containing compound significantly elevated pentoxyresorufin O-dealkylase activity. 5. Overall, mouse show a more limited response in the range of drug-metabolizing enzymes induced by N-heterocycles compared with rat, but as in rat, cytochrome P450 was largely unaffected. PMID- 9849643 TI - Biotransformation of an antihypertensive arylalkylamine analogue in the rat. AB - 1. The excretion and metabolism of N-[2-(3,4-dimethoxyphenyl)ethyl]-5-methoxy N,alpha-dimethyl-2-(phenyl ethynyl) benzenepropanamine (RWJ-26240) in the Wistar rat has been investigated after a single oral dose of 14C-RWJ-26240 (50 mg/kg free base). 2. Plasma samples were obtained for 24 h after dosing and urine and faecal samples were collected over 8 days, and they accounted for 0.9 and 96% of the dose, respectively. 3. Representative samples of plasma, urine and faecal samples were purified for metabolite isolation and identification using HPLC, tlc, mass spectra (CI and EI), 1H-NMR and derivatization. 4. Unchanged RWJ-26240 plus 11 metabolites were identified and accounted for > 80% of the sample radioactivity. 5. Four metabolic pathways for RWJ-26240 are proposed; namely (1) N-demethylation, (2) O-demethylation, (3) phenyl hydroxylation and (4) N dealkylation. Pathways 1-3 appeared to be quantitatively more important. PMID- 9849644 TI - Intermediary metabolism of pentachloronitrobenzene in the control and germ-free rat and rat with cannulated bile ducts. AB - 1. Nearly 70% of single oral doses of 14C-labelled pentachloronitrobenzene (PCNB) was excreted in bile within 24 h. 2. The characterized biliary metabolites of PCNB were either mercapturic acid pathway metabolites or catabolites thereof (thiols, methylthiols, S-glucuronides). 3. A major biliary metabolite was S (aminotetrachlorophenyl)glutathione. 4. Conjugation with glutathione with subsequent catabolism to bis-methylthiotetrachlorobenzene was the major pathway in the control rat. 5. Germ-free experiments showed that only nitro- group displacement occurred, and no nitro- group reduction was detected. PMID- 9849645 TI - Biotransformation of 2-chloroaniline in the Fischer 344 rat: identification of urinary metabolites. AB - 1. The biotransformation of a single i.p. dose of [14C]2-chloroaniline (1.0 mmol/kg, approximately 60 microCi/rat) was investigated in the urine and faeces of the male Fischer 344 rat. 2. During 24 h, 53.1% of the administered radioactivity was eliminated into the urine, while < 1% of the radioactivity appeared in the faeces. 3. The major biotransformation pathways were para hydroxylation and sulphate conjugation. 4-Amino-3-chlorophenyl sulphate was the major urinary metabolite comprising 31.6% of total urinary radioactivity. The para-hydroxylated metabolite, 4-amino-3-chlorophenol (10.8%), and its O glucuronide conjugate (3.7%) were also urinary metabolites. The formation of direct conjugates of 2-chloroaniline, the N-sulphate and N-glucuronide, was significant with as much as 18.6 and 8.6%, respectively, of these metabolites excreted in the urine. The parent compound, 2-chloroaniline, accounted for 16.9% of urinary radioactivity. 4. N-Acetylated products were minor metabolites present in urine as 2-chloro-4-hydroxyacetanilide and its sulphate or glucuronide conjugate. Neither 2-chloroacetanilide nor its oxidation products, 2 chloroglycolanilide and 2-chlorooxanilic acid, were urinary metabolites. PMID- 9849646 TI - Formation in vitro of an inhibitory cytochrome P450 x Fe2+-metabolite complex with roxithromycin and its decladinosyl, O-dealkyl and N-demethyl metabolites in rat liver microsomes. AB - 1. Roxithromycin and its major metabolites found in rat and human urine, namely the decladinosyl derivative (M1), O-dealkyl derivative (M2) and N-demethyl derivative (M3), were incubated with rat liver microsomes and formation of an inhibitory cytochrome P450 (CYP)-metabolite complex and of formaldehyde (measurement of N-demethylation) were determined in vitro. Troleandomycin and erythromycin were also used for comparison. 2. Dexamethasone very significantly induced the microsomal N-demethylations of these macrolide antibiotics. The order of magnitude for the Vmax/Km ratio of N-demethylations by liver microsomes from dexamethasone-treated rats was troleandomycin > erythromycin = M2 > roxithromycin > M3, M1. 3. Formation of an inhibitory P450 x Fe2+-metabolite complex was detected on incubation of these macrolide antibiotics with rat liver microsomes in the presence of an NADPH-generating system and the order of maximum complex formation was troleandomycin > erythromycin > M2 > roxithromycin > M3 > M1. 4. Troleandomycin, erythromycin and M2 inhibited CYP3A-dependent testosterone 6beta hydroxylation catalysed by liver microsomes from the dexamethasone-treated rat by 54, 33 and 23%, respectively, but roxithromycin, M3 and M1 were very weak by comparison. In the untreated rat, only testosterone 6beta-hydroxylation, but not testosterone 16alpha- and 2alpha-hydroxylation and androstenedione formation, activities were inhibited, indicating that inhibitory actions of these antibiotics are specific for CYP3A enzymes in liver microsomes. 5. These results support the view that formation of an inhibitory P450-metabolite complex is prerequisite for the inhibition of CYP3A-dependent substrate oxidations by rat liver microsomes and that M2 (and M3, to a lesser extent) may be the active metabolite that can form an inhibitory P450-metabolite complex by CYP3A enzyme(s). PMID- 9849647 TI - Disposition of methyl ethyl ketoxime in the rat after oral, intravenous and dermal administration. AB - 1. The disposition of 14C-methyl ethyl ketoxime (MEKO) was determined in the male F344 rat following oral, intravenous (i.v.) and dermal administration. 2. Oral doses of 2.7, 27 and 270 mg/kg were primarily excreted as CO2 (71-49%) in decreasing percentage as the dose increased. Excretion in urine (13-26%) and as volatiles (5-18%) increased as the dose increased. Five to 6% of the dose remained in the major tissues after 72 h. 3. An i.v. dose of 2.7 mg/kg was also principally excreted as CO2 (48.8%) with excretion in urine and as expired volatiles accounting for 21.4 and 11.4%, respectively. About 7% of the administered radioactivity remained in the tissues after 72 h. 4. Following dermal administration, 13 and 26% of a 2.7 and 270 mg/kg dose, respectively, were absorbed. Volatilization from the dose site prior to placement in the metabolism cage may account for the low absorption. 5. MEKO was biotransformed to at least five polar metabolites that could only be partially resolved by anion exchange chromatography. Incubation with glucuronidase, but not sulphatase, changed the urinary metabolic profile. Methyl ethyl ketone was a major component in the volatiles. PMID- 9849648 TI - Anti-inflammatory activity of D-002: an active product isolated from beeswax. AB - D-002 is a natural mixture of high molecular weight alcohols isolated and purified from beeswax, which contains triacontanol among its main components. This study was undertaken to investigate the anti-inflammatory effects of D-002 administered by the oral route in two animal models commonly used in the pharmacological screening of anti-inflammatory drugs. D-002 administered orally to rats (100 and 200 mg/kg) produced a mild but significant reduction of exudate volume in carrageenan-induced pleuritic inflammation that was accompanied by a marked and significant decrease of leukotriene B4 (LTB4) levels in the exudate. D 002 (25, 50 and 200 mg/kg) also significantly diminished the granuloma weight in the cotton pellet granuloma in rats. In both cases, D-002 was less effective than indomethacin, which was used as an established anti-inflammatory reference drug. On the other hand, D-002 administered from 25-1000 mg/kg did not induce erosions or gastromucosal lesions in rats, which differs from results usually obtained with non steroidal anti-inflammatory drugs. These results indicate that D-002 is a mild anti-inflammatory agent without any ulcerogenic effect associated. The results suggest that these effects are probably not mediated through an inhibition of cyclooxygenase, but a reduction in LTB4 levels induced by D-002 could explain these results. PMID- 9849649 TI - Essential fatty acid status in cell membranes and plasma of patients with primary Sjogren's syndrome. Correlations to clinical and immunologic variables using a new model for classification and assessment of disease manifestations. AB - In 41 primary Sjogren's syndrome patients we compared fatty acid levels within erythrocyte phospholipids, plasma phospholipids, plasma triglycerides and plasma cholesterol esters, with the immunopathological and clinical disease status. Docosahexaenoic acid was the essential fatty acid (EFA), the levels of which correlated (inversely) most closely with the clinical disease status (r=-0.33 to 0.50). Levels of dihomogammalinolenic acid and eicosapentaenoic acid correlated inversely to levels of IgM rheumatoid factors (r=-0.33) and anti-SSA/Ro antibodies (r=-0.40) respectively. Moreover, levels of anti-SSA/Ro antibodies (r= 0.34-0.40) correlated with levels of the proinflammatory arachidonic acid. Sigma n-3 EFA/sigma n-6 EFA ratios correlated significantly to the quantitative estimates of immunopathological and clinical disease status. Our data are in agreement with current understanding of pro- and anti-immunoinflammatory roles within EFA metabolism, and support the rationale for intervention studies. PMID- 9849650 TI - Comparison of the fatty acid compositions in intraepithelial and infiltrating lesions of the cervix: part I, total fatty acid profiles. AB - In the first part of this study, the possible role of essential total fatty acids and their metabolites during cervical carcinogenesis was investigated. Since membrane lipids play a key role in cell proliferation and differentiation, disturbances in the fatty acid compositions of cell membranes and the modulation of membrane fatty acid compositions received attention in several in vitro studies. There are, however, no reported studies where the actual total and free (unesterified) fatty acid compositions have been determined during the different stages of cervical carcinogenesis. In part I of this ex vivo study, the total fatty acid compositions of normal tissue, intraepithelial and infiltrating lesions of the cervix were compared. The fatty acid profiles that were determined make it possible to speculate about the metabolic pathways followed during cervical carcinogenesis. Lipids were extracted from biopsies of normal tissue (n=36), cervical intraepithelial lesions (n=47) and infiltrating lesions (n=47). Samples, from which the total fatty acid compositions were determined, were saponified, methylated and analysed by gas liquid chromatography (GLC). Essential fatty acid deficiency (EFAD) in the intraepithelial lesions, compared with normal tissue (linoleic acid, P< 0.01), and infiltrating lesions, compared with intraepithelial lesions (linoleic acid and arachidonic acid, P< 0.01) were observed. High levels of oleic acid were also observed when infiltrating lesions were compared with normal tissue (P< 0.01). This EFAD in cancer cells may result in many defective cell mechanisms. Although there are many risk factors for cervical cancer, the human papilloma virus has emerged over the past decade as the leading candidate to be an aetiological factor. There is ample evidence that human viral infections are associated with reduced levels of linoleic acid and thus participate in the depletion of essential fatty acids in cancer cells. PMID- 9849651 TI - Comparison of the fatty acid compositions in intraepithelial and infiltrating lesions of the cervix: part II, free fatty acid profiles. AB - In the second part of this study, the emphasis is on the free fatty acids during cervical carginogenesis, since they may reflect active cell metabolism during this disease process. Lipids were extracted from biopsies of normal epithelial tissue (N) (n=36), cervical intraepithelial lesions (CIL) (n=47), and infiltrating lesions (Ca) (n=47) of the cervix. Samples, from which the free fatty acid compositions were determined, were saponified, methylated and analysed by GLC. In accordance with results obtained on total fatty acid compositions, essential fatty acid deficiency (EFAD) in the intraepithelial lesions, compared with normal tissue (linoleic acid, P< 0.01), and infiltrating lesions compared with intraepithelial lesions (linoleic acid and arachidonic acid, P< 0.01) were observed. High levels of oleic acid were also observed when infiltrating lesions were compared with normal tissue (P < 0.01). As previously mentioned by us in part I of this study, with regard to possible disturbances in metabolic pathways based on the total fatty acid profiles during stages of cervical cancer, EFAD is prevalent during cervical carcinogenesis. This EFAD in cancer cells may result in many defective cell mechanisms, since fatty acids are associated with biochemical events such as lipid peroxidation, signal transduction and immune responses. The high level of oleic acid in cancer cells is known to activate PKC and thus contribute to the continous growth stimulus thought to exist in malignant cells. From a therapeutic viewpoint, substantial changes in the fatty acid composition of the membranes can be produced in cancer cells by selective fatty acid supplementation strategies. At present, modifications of the fatty acid compositions of cell membranes represent an experimental model that has promoted increased understanding of lipid transportation, membrane remodelling, and the relationship between membrane lipids and membrane function. By addressing factors responsible for insufficient essential fatty acid levels, carginogenesis may be prevented or treated. The clinical feasibility of using modification of fatty acids in tumours or cancer by diet or perfusion as an adjunct to standard therapies should be tested. PMID- 9849652 TI - Comparison of the fatty acid compositions in intraepithelial and infiltrating lesions of the cervix: part III, saturated and unsaturated fatty acid profiles. AB - The purpose of the third part of this study is to construct a basic lipid model (this includes information regarding total and free saturated, monounsaturated and polyunsaturated fatty acid contents, as well as total and free fatty acid saturation and double bond indexes, and comparisons of total and free n-3, n-6, n 7 and n-9 fatty acids in normal epithelial tissue, and intraepithelial and infiltrating lesions of the cervix) which, together with the individual total and free fatty acid profiles given in parts I and II of this study, should provide an understanding of the turnover of total and free acids, especially essential fatty acids, during cervical carcinogenesis. Such information can serve as a sound basis for further studies in an attempt to access this disease process. We observed an increase in monounsaturated fatty acid values in cancer tissue compared with normal tissue and a decrease of saturated fatty acid values in cancer tissue compared with normal tissue. Based on our observations, we speculate that because of the depletion of polyunsaturated fatty acids, monounsaturated fatty acid are synthesized to compensate for this loss; a possible source for the monounsaturated fatty acids are the saturated fatty acids via elongation and/or desaturation. Of particular interest is the n-3 fatty acid docosahexaenoic acid, the most unsaturated lipid in the biological systems, detected in very small amounts only in cancer cells of the cervix. PMID- 9849653 TI - Prostaglandin E2 receptor EP3alpha subtype: the role of N-glycosylation in ligand binding as revealed by site-directed mutagenesis. AB - Functional mouse prostaglandin E2 (PGE2) receptor EP3alpha subtype has been expressed in insect cells using a baculovirus system (Huang C. and Tai H.-H. Biochem J 1995; 307: 493-498). EP3alpha receptor has two potential sites (Asn-X Ser/Thr), Asn 16 and Asn 193, for N-glycosylation. The role of glycosylation in ligand binding of the EP3alpha receptor was investigated by site-directed mutagenesis. Asn was mutated to Gln in each of the two potential glycosylation sites in the EP3alpha receptor. Recombinant wild-type and mutant EP3alpha receptors were expressed in insect cells using baculovirus. Ligand binding assay indicated that the affinity of PGE2 binding was reduced by 50% in the Gln 193 mutant EP3alpha receptor, while the specificity of ligand binding was unaltered. The affinity for PGE2 binding was not affected in the Gln 16 mutant EP3alpha receptor. However, its specificity was partially changed as the EP3-specific agonist became less effective in displacing the [3H]-PGE2 binding to the mutant receptor. These results indicated that N-glycosylation of the EP3alpha receptor could partially affect the affinity and specificity of the ligand binding. PMID- 9849654 TI - Cytokine levels affected by gamma-linolenic acid. AB - This study was undertaken to assess whether gamma-linolenic acid (GLA) in the form of evening primrose oil (EPO) could affect rat serum cytokines, interferon gamma (IFN-gamma), monocyte chemotactic protein-1 (MCP-1) and tumour necrosis factor-alpha (TNF-alpha). The following diets were administered: control, glucan, Freund's adjuvant and glucan plus Freund's adjuvant with and without GLA. In the presence of GLA, the IFN-gamma and MCP-1 levels were significantly decreased in contrast to the control group of TNF-alpha, which was significantly stimulated. On account of interaction between diets and GLA, the remaining diet groups of TNF alpha were either not affected or were inhibited in the presence of GLA. The observations indicate that GLA may modulate the level of serum IFN-gamma, MCP-1 and TNF-alpha, which may be a worthwhile line of treatment in certain human diseases. PMID- 9849655 TI - Role of endogenous endothelin peptides in intestinal ischemia-reperfusion injury in rats. AB - In this study, the effects of BQ123 (an ET(A) receptor antagonist), bosentan (a nonselective ET(A)-ET(B) antagonist), and phosphoramidon (an endothelin converting enzyme inhibitor) were investigated on intestinal mucosal lesion formation and changes in tissue PGE2 and LTC4 levels due to intestinal ischemia reperfusion (I/R) injury in rats. Following 30 min of ischemia, the substances were given via the inferior caval vein, and 10 min later the intestine was subjected to reperfusion for 30 min. The intestinal specimens were evaluated both microscopically and the tissue PGE2 and LTC4 levels were obtained for each group. The histopathologic examination revealed a significant reduction in tissue injury in both BQ123 and phosphoramidon pretreated groups compared with the control group. Bosentan, on the contrary, did not decrease the injury. The pharmacologic examination revealed a significant reduction of PGE2-like activity in both BQ123 and phosphoramidon pretreated groups, compared with the control group, while LTC4 like activity remained unchanged except for an increase in the bosentan pretreated group. PMID- 9849656 TI - Comparative anti-mitotic effects of lithium gamma-linolenate, gamma-linolenic acid and arachidonic acid, on transformed and embryonic cells. AB - The effects of gamma-linolenic acid (GLA), the lithium salt of gamma-linolenic acid (LiGLA) and arachidonic acid (AA) were compared at doses of 50 microg/ml for periods of 6 and 24 h on cell cycle progression and apoptosis induction in transformed and in normal cells. In WHCO3 (oesophageal cancer) cells and on primary embryonic equine lung cells, we found LiGLA to be the most effective in apoptosis induction. After 24 h, 94% of the WHCO3 cancer cells and 44% of the primary embryonic equine lung cells exposed to LiGLA were apoptotic. The WHCO3 cancer cells were also very susceptible to the apoptosis-inducing effects of AA (56%) and GLA (44%), whereas the embryonic equine lung cells were much less affected by these two fatty acids. After 6 h exposure to all three compounds, most of the cycling WHCO3 cancer cells were blocked in S-phase. After 24 h treatment, some of the S-phase cells exposed to AA and GLA were apparently able to move into the G2/M phase, the LiGLA exposed cells were mostly apoptotic and no cycling cells were present. The primary embryonic equine lung cells were fairly resistant to the cytotoxic effects of GLA and AA. From our studies we conclude that, although LiGLA was the most toxic to the cancer cells, it is apparently less selective, compared to AA and GLA, in the killing of cancer and normal cells. It would also appear that the lithium might have added to the cytotoxic effects of LiGLA. The mechanism needs to be clarified. PMID- 9849657 TI - Neuropharmacology of AMPA and kainate receptors. PMID- 9849658 TI - Pharmacology and toxicology of ATOA, an AMPA receptor antagonist and a partial agonist at GluR5 receptors. AB - (RS)-2-Amino-3-[3-(carboxymethoxy)-5-tert-butyl-4-isoxazolyl]propi onic acid (ATOA) has previously been described as an antagonist at (RS)-2-amino-3-(3 hydroxy-5-methyl-4-isoxazolyl)propionic acid (AMPA) receptors with an IC50 value of 150 microM towards AMPA-induced depolarisation in the rat cortical wedge preparation. ATOA has now been shown also to be a partial agonist at recombinant GluR5 receptors, expressed in Xenopus oocytes, with an EC50 value of 170 microM and a relative efficacy of 0.17 +/- 0.04 compared with responses produced by kainic acid (1.0). Using cultured cerebral cortical neurones as a test system and leakage of lactate dehydrogenase (LDH) as an indicator of cell damage, ATOA was shown to be cytotoxic (ED50 > 300 microM), though much less toxic than the structurally related dual AMPA and GluR5 agonist, (RS)-2-amino-3-(3-hydroxy-5 tert-butyl-4-isoxazolyl)propionic acid (ATPA) (ED50 = 14 +/- 2 microM). The toxic effect of ATPA was sensitive to 6,7-dinitroquinoxaline-2,3-dione (DNQX) but was not significantly reduced by the selective AMPA receptor antagonist, (RS)-2-amino 3-[3-(carboxymethoxy)-5-methyl-4-isoxazolyl]propionic acid (AMOA). The toxicity of ATOA (1 mM) could not be significantly attenuated by co-administration of AMOA (300 microM) or DNQX (25 microM). A structure-activity analysis indicates that the tert-butyl group of ATPA and ATOA facilitates the interaction of these compounds with GluR5 receptors. PMID- 9849659 TI - Decahydroisoquinolines: novel competitive AMPA/kainate antagonists with neuroprotective effects in global cerebral ischaemia. AB - In the present studies, we have evaluated the activity of a series of glutamate receptor antagonists from the decahydroisoquinoline group of compounds both in vitro and in vivo. Compound activity at alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (AMPA) and kainate receptors was assessed using ligand binding to cloned iGluR2 and iGluR5 receptors and on responses evoked by AMPA and N-methyl-D aspartate (NMDA) in the cortical wedge preparation. In vivo, compounds were examined for antagonist activity electrophysiologically in the rat spinal cord preparation and in the gerbil model of global cerebral ischaemia. Compounds tested were LY293558, which has been shown to protect in models of focal cerebral ischaemia, LY202157 (an NMDA antagonist), LY246492 (an NMDA and AMPA receptor antagonist), LY302679, LY292025, LY307190, LY280263, LY289178, LY289525, LY294486 (AMPA/kainate antagonists) and LY382884 (an iGluR5 selective antagonist). Results obtained support a role for AMPA receptors in cerebral ischemia. LY377770 (a mixed AMPA/iGluR5 antagonist and active isomer of LY294486) demonstrated good neuroprotection with a 2-h time window and may therefore be useful in the treatment of ischaemic conditions. PMID- 9849660 TI - Pharmacological differentiation of kainate receptors on neonatal rat spinal motoneurones and dorsal roots. AB - The objectives of this study, conducted on neonatal rat spinal cord and dorsal roots in vitro, were to characterise the actions of a range of willardiine analogues on GluR5-containing kainate receptors present in dorsal roots, to determine whether GluR5-containing receptors are also present on motoneurones, and to differentiate responses mediated by kainate receptors from those mediated by AMPA receptors on motoneurones. (S)-5-Trifluoromethyl-willardiine, (S)-5 iodowillardiine, (S)-5-iodo-6-azawillardiine and ATPA were found to be potent agonists of kainate receptors on dorsal roots (EC50 values 0.108 +/- 0.002, 0.127 +/- 0.010, 0.685 +/- 0.141 and 1.3 +/- 0.3 microM, respectively) being more potent but of lower efficacy than kainate (EC50 value 14.8 +/- 1.8 microM). (S)-5 Iodo-6-azawillardiine blocked kainate-induced depolarisations of the dorsal root, probably via its desensitising action. Kainate-induced responses of dorsal roots were weakly antagonised by (RS)-3,5-dicarboxyphenylglycine (DCPG) (apparent KD 1.5 +/- 0.4 mM). Kainate receptors containing GluR5 subunits do not appear to be present on motoneurones since (RS)-3,5-DCPG (1 mM) potentiated rather than antagonised kainate-induced depolarisations of motoneurones. Although (S)-5 iodowillardiine (a potent and selective agonist at GluR5-containing kainate receptors) depolarised motoneurones (EC50 value 5.8 +/- 0.6 microM), such depolarisations were antagonised by both (RS)-3,4- and (RS)-3,5-DCPG, which are selective AMPA receptor antagonists at motoneurones, showing a KD value of 73 microM (Schild slope, 0.96 +/- 0.09) and an apparent KD value of 123 +/- 38 microM, respectively. This accords with the previously reported activity of willardiine analogues at AMPA receptors. Since neither (RS)-3,4- nor (RS)-3,5 DCPG antagonised kainate-induced motoneuronal depolarisations but cyclothiazide enhanced and GYK153655 blocked these responses it is possible that a component of the kainate response may be mediated by a population of DCPG-insensitive AMPA receptors on motoneurones. However, it is also possible that a population of kainate receptors other than those containing GluR5 subunits, are responsible for these effects. The new compounds introduced in this study are likely to be useful tools for studying the physiological role of kainate receptors in CNS function. PMID- 9849661 TI - Antagonism of neuronal kainate receptors by lanthanum and gadolinium. AB - The effects of lanthanum and gadolinium on currents evoked by excitatory amino acids were studied in cultured rat hippocampal and cortical neurons, in freshly dissociated dorsal root ganglion neurons, and in human embryonic kidney 293 cells expressing the GluR6 kainate receptor subunit. In all of these cells, currents mediated by kainate-preferring receptors were antagonized by low micromolar concentrations of the trivalent ions. At negative holding potentials, the IC50 values for inhibition in DRG cells were 2.8 microM for La and 2.3 microM for Gd. Kainate receptor-mediated currents in hippocampal neurons and in 293 cells expressing GluR6 were blocked by La with IC50 values of 2.1 and 4.4 microM, respectively. Steady-state inhibition by the lanthanides showed very slight dependence on membrane potential, however, we were not able to resolve any systematic variation with membrane potential in the kinetics of block onset or recovery. Inhibition was not use-dependent and was not overcome by increasing the concentration of agonist. These results indicate that lanthanides probably do not bind deep within the ion pore or directly compete for the agonist binding site. In contrast to neuronal AMPA receptors, which require more than 100 microM lanthanides for half-maximal blockade, the inhibition of neuronal and recombinant kainate receptors by these ions displays significantly higher potency. PMID- 9849662 TI - Activation and desensitization properties of native and recombinant kainate receptors. AB - The activation-inactivation properties of membrane currents induced by the rapid application of glutamate or kainate were studied in cultured hippocampal neurons and in HEK cells transfected with a cDNA encoding the GluR6 subunit. The onset of desensitization was rapid and similar in native and recombinant channels (approximately 80 s(-1) of onset rate constant). Recovery from desensitization was slow and agonist-dependent in neurons, proceeding slightly faster in GluR6 receptors. Half-maximal activation (EC50) of native channels was obtained at a glutamate concentration of 330 microM, while the half-maximal steady state desensitization (IC1/2) was attained at 2.8 microM. These values differed from those obtained in recombinant receptors (EC50 = 762 microM and IC1/2 = 0.44 microM). A small window under the crossing point of activation and inactivation curves was observed, indicating that, for some concentrations of either agonist, steady state channel activity could exist. In native receptors, this window presented maximum values at approximately 100 microM for glutamate, which predicted well the potency of glutamate to reduce the GABAergic drive in hippocampal slices. These data indicate that for neuronal kainate receptors, the concentrations for half activation and half inactivation differ by two orders of magnitude such that the maximum response to a maintained concentration of glutamate is small, and the steady state dose response curve is skewed and bell shaped. PMID- 9849663 TI - LY339434, a GluR5 kainate receptor agonist. AB - The activity of a gamma-substituted glutamate analogue, (2S, 4R, 6E)-2-amino-4 carboxy-7-(2-naphthyl)hept-6-enoic acid (LY339434) and (2S,4R)-4-methylglutamic acid at ionotropic glutamate receptors has been examined. Ligand binding studies were performed using [3H] AMPA binding to membranes expressing either homomeric recombinant GluR1, GluR2, GluR4 receptors, and [3H] kainate binding to GluR5 and GluR6 kainate receptors. LY339434 and (2S,4R)-4-methylglutamic acid showed selectivity in ligand binding studies for kainate receptors over AMPA receptors. Within the kainate class of glutamate receptors, LY339434 showed selectivity for GluR5 over GluR6 whereas (2S,4R)-4-methylglutamic acid showed high affinity for both GluR5 and GluR6 kainate receptors. Examination of the functional activity of LY339434 and (2S,4R)-4-methylglutamic acid showed that both compounds evoked inward currents in dorsal root ganglion neurons (DRG) with estimated EC50 values of 0.8 +/- 0.2 microM and 0.17 +/- 0.04 microM, respectively. In GluR5 expressing HEK 293 cells, LY339434 evoked inward currents with an estimated EC50 value of 2.5 +/- 0.9 microM but had little effect on GluR6 expressing cells at concentrations less than 100 microM. LY339434 was a weak AMPA receptor agonist (EC50 values > 300 microM) as determined by activity in acutely isolated cerebellar Purkinje neurons. LY339434 and (2S,4R)-4-methylglutamic acid had agonist activity at NMDA receptors studied in cultured hippocampal neurons with EC50s of 2.5 microM and 11.7 microM, respectively. These results indicate that both LY339434 and (2S,4R)-4-methyl glutamic acid may be useful pharmacological tools for the examination of kainate receptors. PMID- 9849664 TI - The GluR5 subtype of kainate receptor regulates excitatory synaptic transmission in areas CA1 and CA3 of the rat hippocampus. AB - Activation of kainate receptors depresses excitatory synaptic transmission in the hippocampus. In the present study, we have utilised a GluR5 selective agonist, ATPA [(RS)-2-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl)propanoic acid], and a GluR5 selective antagonist, LY294486 [(3SR,4aRS,6SR,8aRS)-6-([[(1H-tetrazol-5-y l)methyl]oxy]methyl)-1,2,3,4,4a,5,6,7,8,8a-decahydroisoquinoline-3 -carboxylic acid], to determine whether GluR5 subunits are involved in this effect. ATPA mimicked the presynaptic depressant effects of kainate in the CA1 region of the hippocampus. It depressed reversibly AMPA (alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid) receptor-mediated field excitatory postsynaptic potentials (field EPSPs) with an IC50 value of approximately 0.60 microM. The dual-component excitatory postsynaptic current (EPSC) and the pharmacologically isolated NMDA (N-methyl-D-aspartate) receptor-mediated EPSC were depressed to a similar extent by 2 microM ATPA (61 +/- 7% and 58 +/- 6%, respectively). Depressions were associated with an increase in the paired-pulse facilitation ratio suggesting a presynaptic locus of action. LY294486 (20 microM) blocked the effects of 2 microM ATPA on NMDA receptor-mediated EPSCs in a reversible manner. In area CA3, 1 microM ATPA depressed reversibly mossy fibre-evoked synaptic transmission (by 82 +/- 10%). The effects of ATPA were not accompanied by any changes in the passive properties of CA1 or CA3 neurones. However, in experiments where K+, rather than Cs+, containing electrodes were used, a small outward current was observed. These results show that GluR5 subunits comprise or contribute to a kainate receptor that regulates excitatory synaptic transmission in both the CA1 and CA3 regions of the hippocampus. PMID- 9849665 TI - GluR5 kainate receptor mediated synaptic transmission in rat basolateral amygdala in vitro. AB - A non-NMDA and non-AMPA receptor mediated excitatory synaptic response was identified in intracellularly recorded basolateral amygdala (BLA) neurons in an in vitro slice preparation. Synaptic potentials were evoked by stimulation of either the external capsule (EC) or basal amygdala (BA). NMDA and GABA(A) receptors were blocked by inclusion of 100 microM (+/-)-2-amino-5 phosphonopentanoic acid and 10 microM bicuculline in the perfusion solution. The AMPA receptor-selective allosteric antagonists GYKI 52466 (50 microM) and GYKI 53655 (50 microM) partially suppressed depolarizing synaptic responses evoked by single shock EC stimulation, but fully blocked synaptic responses evoked by BA stimulation. In recordings carried out in the presence of the AMPA receptor antagonists, EC stimulation with pulse trains (5-8 pulses at 50-100 Hz) evoked a large increase in the amplitude of synaptic responses. The AMPA receptor independent component of the train-induced synaptic response had a null potential near 0 mV. Such AMPA receptor-independent, train-evoked synaptic responses were largely blocked by the AMPA/kainate receptor antagonist 6-cyano-7 nitroquinoxaline-2,3-dione (20 microM; 85 +/- 4%). In addition, the responses were blocked by the GluR5-selective kainate receptor antagonist LY293558 (10 microM; 95 +/- 2%). These results indicate that a component of the EC (but not the BA) synaptic response is mediated by kainate receptors containing the GluR5 subunit. PMID- 9849666 TI - Actions of kainate and AMPA selective glutamate receptor ligands on nociceptive processing in the spinal cord. AB - Kainate receptors expressing the GluR5 subunit of glutamate receptor are present at high levels on small diameter primary afferent neurones that are considered to mediate nociceptive inputs. This suggests that GluR5 selective ligands could be novel analgesic agents. The role of kainate receptors on C fibre primary afferents has therefore been probed using three compounds that are selective for homomeric GluR5 receptors. The agonist, ATPA, and the antagonists, LY294486 and LY382884, have been tested in four models of nociception: responses evoked by noxious stimulation of the periphery have been recorded electrophysiologically (1) from hemisected spinal cords from neonatal rats in vitro, (2) from single motor units in adult rats in vivo, (3) from dorsal horn neurones in adult rats in vivo, and (4) in hotplate tests with conscious mice. In some protocols comparisons were made with the AMPA selective antagonist GYKI 53655. The agonist ATPA reduced nociceptive reflexes in vitro, but failed to have effects in vivo. In all tests, the GluR5 antagonists reduced nociceptive responses but only at doses that also affected responses to exogenous AMPA. The AMPA antagonist reduced nociceptive responses at doses causing relatively greater reductions of responses to exogenous AMPA. The results indicate that GluR5 selective ligands do reduce spinal nociceptive responses, but they are not strongly analgesic under these conditions of acute nociception. PMID- 9849667 TI - Interactions of GYKI 52466 and NBQX with cyclothiazide at AMPA receptors: experiments with outside-out patches and EPSCs in hippocampal neurones. AB - In outside-out patches from cultured hippocampal neurones, glutamate (1 mM) applied for 1 ms evoked currents which rose rapidly (tau(on) 451 +/- 31 micros) to a peak and then deactivated with slower kinetics (1.95 +/- 0.13 ms). Offset time constants were significantly slower with longer application durations (tau(off) 3.10 +/- 0.19, 3.82 +/- 0.25, 4.80 +/- 0.65 and 7.56 +/- 0.65 ms with 10, 20, 100 and 500 ms applications respectively). Desensitization was complete within 100 ms with a similar rate for all application durations (4.74 +/- 0.34 ms with 100 ms applications). GYKI 52466 reduced inward peak currents with an IC50 of 11.7 +/- 0.6 microM and had similar potency on steady-state currents to longer glutamate applications. GYKI 52466 had no significant effect on desensitization or deactivation time constants but caused a modest and significant prolongation of onset kinetics at higher concentrations. Cyclothiazide (100 microM) potentiated steady-state currents 25-fold at 100 ms and caused a modest but significant slowing in onset kinetics (601 +/- 49 micros with 1 ms applications) but a more pronounced prolongation of deactivation time constants (5.55 +/- 0.66 ms with 1 ms applications). In 50% of neuronal patches cyclothiazide completely eliminated desensitization. In those patches with residual desensitization, the rate was not significantly different to control (5.36 +/- 0.43 ms with 100 ms applications). Following 100 ms applications of glutamate, GYKI 52466 had IC50s of 11.7 +/- 1.1 microM and 75.1 +/- 7.0 microM in the absence and presence of cyclothiazide (100 microM) respectively. Onset kinetics were slowed from 400 +/- 20 micros to 490 +/- 30 micros by cyclothiazide (100 microM) and then further prolonged by GYKI 52466 (100 microM) to a double exponential function (tau(on1) 1.12 +/- 0.13 ms and tau(on2) 171.5 +/- 36.5 ms). GYKI 52466 did not re-introduce desensitization but concentration-dependently weakened cyclothiazide's prolongation of deactivation time constants (1 ms applications: 5.01 +/- 0.71, 4.47 +/- 0.80 and 2.28 +/- 0.64 ms with GYKI 52466 30, 100 and 300 microM respectively). NBQX reduced peak current responses with an IC50 of 28.2 +/- 1.3 nM. Paradoxically, steady-state currents with 500 ms applications of glutamate were potentiated from 3.3 +/- 1.2 pA to 29.4 +/- 6.4 pA by NBQX (1 nM). Higher concentrations of NBQX then antagonized this potentiated response. The potency of NBQX in antagonizing steady-state currents to 500 ms applications of glutamate (IC50 120.9 +/- 30.2 nM) was 2-fold less than following 100 ms applications (IC50 67.7 +/- 2.6 nM). NBQX had no effect on rapid onset, desensitization or deactivation time constants. However, a slow relaxation of inhibition was seen with longer applications. NBQX was 2-5-fold less potent against inward currents in the presence of cyclothiazide (100 microM) depending on the application duration but had no effect on the rapid onset, desensitization or deactivation time constants. The same relaxation of inhibition was seen as with NBQX alone. NBQX (1 microM) reduced AMPA receptor-mediated EPSC amplitude to 7 +/- 1% of control with no effect on kinetics. Cyclothiazide (330 microM) caused a 2.8-fold prolongation of the decay time constant (control 26.6 +/- 2.2 ms, cyclothiazide 74.2 +/- 7.6 ms, n = 9). Additional application of NBQX (1 microM) partly reversed this prolongation to 1.9 fold (47.7 +/- 2.5 ms, n = 5). These results support previous findings that cyclothiazide also allosterically influences AMPA receptor agonist/antagonist recognition sites. There were no interactions between NBQX and cyclothiazide on desensitization or deactivation time constants of glutamate-induced currents but clear interactions on EPSC deactivation kinetics. This raises the possibility that the interactions of NBQX, GYKI 52466 and cyclothiazide on AMPA-receptor-mediated EPSC kinetics observed are due to modulation of glutamate-release at presynaptic AMPA receptors. PMID- 9849668 TI - Variations in the tangential distribution of postsynaptic glutamate receptors in Purkinje cell parallel and climbing fiber synapses during development. AB - Major factors affecting the responses of a neuron to release of glutamate include the kinds and distribution of glutamate receptors in the neuron and their distribution along the surface of the postsynaptic membrane (tangential distribution). The latter distribution pattern is established during the development of the synapse and could be modified during maturation of synapse structure/function and through adult synapse plasticity. Parallel and climbing fiber synapses of cerebellar Purkinje cells are good models for studying this pattern because they contain two major kinds of ionotropic glutamate receptors, alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and delta, that are involved in adult plasticity and show differences in distribution, and because these two synapse types show complex changes in architecture and glutamate receptor distributions during development. In the present study, both AMPA and delta receptors showed variations in tangential distributions during many stages of development from postnatal day 2 to adult; i.e. qualitative assessment showed that receptors are concentrated either near the center or in outer portions of the synapse, while they are rare or absent from the perisynaptic region. Quantitative analysis showed statistically significant nonuniformities at some ages; the most common nonuniformity in these cases appears to be a drop-off in receptor density in the outer 20% of the synapse. Statistical analyses also indicated that distribution patterns did not change significantly with age. In contrast to the ionotropic receptors, the metabotropic glutamate receptor, mGluR1alpha, was found mainly in the perisynaptic region both during development and in adults. Differences in the distribution of glutamate receptors may be necessary to assure an effective response to glutamate release and may be modified through synaptic plasticity. Our findings show that the basic patterns of distribution of ionotropic and metabotropic glutamate receptors in synapses are established early in development, indicating that the postsynaptic density/membrane region is highly organized even in the immature synapse. PMID- 9849669 TI - Biochemical and immunocytochemical characterization of GRIP, a putative AMPA receptor anchoring protein, in rat brain. AB - The mechanisms by which glutamate receptors are concentrated in brain excitatory synapses are believed to involve interactions between receptor subunits and postsynaptic anchoring or scaffolding proteins. Recently GRIP, a protein containing seven PDZ domains, was identified as an AMPA receptor binding protein and implicated in the synaptic targeting of AMPA receptors. Here we show that GRIP mRNA is also expressed in some tissues outside of the brain, including testis and kidney. Specific antibodies were raised to study GRIP protein. On Western blots, GRIP protein appears as a heterogeneous band (approximately 130 kilodaltons) which is expressed in widespread brain regions and throughout postnatal development. Biochemical studies reveal that GRIP is largely membrane associated and enriched in the postsynaptic density (PSD), though not as highly concentrated in the PSD as is PSD-95. By immunohistochemistry, GRIP is distributed in a somatodendritic pattern in neurons of adult rat brain, with especially prominent expression in a subset of interneurons. PMID- 9849670 TI - Surface expression and metabolic half-life of AMPA receptors in cultured rat cerebellar granule cells. AB - The surface expression and metabolic turnover time of alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate (AMPA) receptors are important factors in determining the involvement of these proteins in synaptic function. We used the membrane-impermeant chemical crosslinking reagent BS3 and subsequent immunoblot analysis to show that a large proportion of each of the AMPA receptor subunits GluR1-4 is surface expressed in cultured rat cerebellar granule cells. The apparent molecular mass of the individual subunits was 105 kDa and the surface expressed crosslinked complex was 530 kDa for each of the antibodies tested. These results are consistent with functional ionotropic glutamate receptors being pentameric subunit assemblies. We investigated the time course of AMPA receptor surface expression using the membrane-impermeant biotinylating agent NHS-SS biotin. The half-life of surface expressed AMPA receptors was found to be 30 h. To estimate the mean degradation rate of AMPA receptors and total trichloracetic acid (TCA)-precipitable protein we used [35S]methionine/cysteine pulse-chase labelling. The half-life of AMPA receptors immunoprecipitated with anti-GluR1 antibody was approximately 48 h and the half-life of total TCA-precipitable protein from the same samples was 37 h. PMID- 9849671 TI - Assembly intracellular targeting and cell surface expression of the human N methyl-D-aspartate receptor subunits NR1a and NR2A in transfected cells. AB - The intracellular trafficking, assembly, and cell surface targeting of the human N-methyl-D-aspartate receptor subunits NR1a and NR2A has been studied using both transiently and permanently transfected mammalian cell lines. The expression of either NR1a or NR2A alone does not result in significant cell surface expression of either subunit as determined by cell surface biotinylation and immunofluorescence staining. When NR1a is expressed alone large intracellular accumulations of the subunit are formed which do not co-localize with the golgi apparatus markers protein p58 and wheat germ agglutinin, but do co-localize with the endoplasmic reticulum marker calreticulin. Co-expression of NR1a and NR2A results in a reduction of these intracellular accumulations and the appearance of both subunits on the cell surface. Immunoprecipitation of NR1a from in vitro translated subunit proteins showed that NR2A could only be immunoprecipitated with NR1a when both subunits were co-synthesized in the presence of microsomes. When cells expressing NR1a and NR2A were incubated with [35S]methionine in the presence of Brefeldin-A, a drug which prevents protein transport from the endoplasmic reticulum, NR2A could be immunoprecipitated by an antiserum specific for NR1a. Together these results suggest that the NMDA receptor subunits are assembled in the endoplasmic reticulum and that co-synthesis of the subunits is necessary for their association and their successful cell surface targeting. PMID- 9849672 TI - NMDA receptor diversity in the cerebellum: identification of subunits contributing to functional receptors. AB - Recent studies of N-methyl-D-aspartate (NMDA) receptors have led to the suggestion that there are two distinct classes of native NMDA receptors, identifiable from their single-channel conductance properties. 'High-conductance' openings arise from NR2A- or NR2B-containing receptors, and 'low-conductance' openings arise from NR2C- or NR2D-containing receptors. In addition, the low conductance channels show reduced sensitivity to block by Mg2+. The readily identified cell types and simple architecture of the cerebellum make it an ideal model system in which to determine the contribution of specific subunits to functional NMDA receptors. Furthermore, mRNA for all of these four NR2 subunits are represented in this brain region. We have examined NMDA channels in Purkinje cells, deep cerebellar nuclei (DCN) neurons and Golgi cells. First we find that NR2D-containing NMDA receptors give rise to low-conductance openings in cell attached recordings from Purkinje cells. The characteristic conductance of these events cannot, therefore, be ascribed to patch excision. Second, patches from some DCN neurons exhibit mixed populations of high- and low-conductance openings. Third, Golgi cells also exhibit a mixed population of high- and low-conductance NMDA receptor openings. The features of these low-conductance openings are consistent with the presence of NR2D-containing NMDA receptors, as suggested by in situ hybridization data. On the other hand the existence of high-conductance channels, with properties typical of NR2B-containing receptors, was not expected. Our results provide new evidence about the subunit composition of NMDA receptors in identified cerebellar cells, and suggest that examination of single-channel properties is a potentially powerful approach for determining the possible subunit composition of native NMDA receptors. PMID- 9849673 TI - The role of hydrophobic interactions in binding of polyamines to non NMDA receptor ion channels. AB - Block of kainate subtype glutamate receptor channels by internal polyamines was analysed using outside out patches from HEK 293 cells transiently transfected with GluR6(Q). Tetramines with different numbers and spacing of methylene groups between NH2 groups produced biphasic rectification well fit by the Woodhull model for a weakly permeable ion channel blocker. Such analysis revealed an increase in binding energy of 611 cal M(-1) for each methylene group added over the range 6 12 (CH2), suggesting that a major component of block by polyamines involves hydrophobic binding. Isomers with the same number of CH2 groups but different spacing between NH2 groups showed similar affinity. Due to differences in pKa values for protonation of NH2 groups, the average charge on the tetramines studied would be expected to vary from 3.98 to 2.22 at physiological pH; despite this, the voltage dependence of block was similar for all tetramines tested, with a mean value for ztheta of 1.82, similar to values for polyamines with five or six NH2 groups. In contrast, for 1,3-propane diamine (DA3 ztheta 0.83), and the N propyl- (ztheta 1.42) and N,N'-diethyl- (ztheta 1.37) analogues of DA3, there was an increase in the voltage dependence of block on addition of hydrophobic groups. PMID- 9849674 TI - No change in NMDA receptor-mediated response rise-time during development: evidence against transmitter spillover. AB - Glutamatergic transmission was examined in tadpole optic tectum to test the possibility that transmitter concentration reaching N-methyl-D-aspartate (NMDA) receptors increases over development. Pharmacologically isolated NMDA receptor mediated transmission was monitored with whole-cell recordings. Synaptic responses were recorded from cells at different locations in the optic tectum, corresponding to different stages of development. Rise-times and decay-times of NMDA currents were analyzed. We found no significant correlation between rise time and developmental stage. As NMDA rise-times can correlate with concentration for glutamate concentrations below 200 microM, these results argue that, if there is developmental variation in transmitter concentration, this occurs for values greater than 200 microM. Furthermore, we found a correlation between rise-times and decay-times, consistent with a model in which transmitter concentration is high and rise-time is controlled by channel closings. These results argue against synaptic models in which low concentrations of transmitter (as by spillover from nearby release sites) selectively activates NMDA receptors. PMID- 9849675 TI - An investigation of the expression mechanism of LTP of AMPA receptor-mediated synaptic transmission at hippocampal CA1 synapses using failures analysis and dendritic recordings. AB - There is considerable controversy surrounding the mechanism of expression of long term potentiation of AMPA receptor-mediated synaptic transmission in the CA1 region of the hippocampus, a process thought to be important for learning and memory in the mammalian CNS. We have re-examined the expression mechanism of this form of synaptic plasticity using whole-cell dendritic recordings, minimal stimulation to activate one or a few synapses, and failures analysis. Dendritic recordings provide improved resolution of small synaptic events, as compared to previous studies using somatic recordings, because there is less dendritic filtering of signals. We find that long-term potentiation (LTP) is associated with changes in the size of synaptic responses when they occur (potency) in all cells and this is accompanied by significant decreases in failure rate in approximately 60% of the experiments. This suggests that in some cells an increase in quantal amplitude is the sole expression mechanism for LTP and, in the cells where failure rate decreased, there is an additional mechanism causing a change in quantal content. PMID- 9849676 TI - Candidate editases for GluR channels in single neurons of rat hippocampus and cerebellum. AB - RNA editing by site selective adenosine deamination changes codons in several nuclear transcripts in the mammalian brain and affects critical properties of the encoded proteins, as exemplified by the calcium permeability of AMPA receptor channels. The recently cloned RNA dependent adenosine deaminases ADAR1, ADAR2 and ADAR3 form a small family of sequence-related candidate editases which are expressed in brain and other tissues at distinct levels and patterns. We have employed single-cell polymerase chain reaction of hippocampal CA1 and CA3 pyramidal neurons and cerebellar Purkinje and Bergmann glial cells in an attempt to evaluate the expression of these enzymes at a cellular level. We found ADAR2 expressed in all cells analyzed; approximately 50% of the cells co-expressed ADAR1 or ADAR3. The differential ADAR expression revealed by our study might underlie the distinct editing efficiencies and selectivities in different GluR subunit transcripts. PMID- 9849677 TI - Micromolar L-glutamate induces extensive apoptosis in an apoptotic-necrotic continuum of insult-dependent, excitotoxic injury in cultured cortical neurones. AB - Excitotoxicity induced by L-glutamate (Glu), when examined in a pure neuronal cortical culture, involved widespread apoptosis at concentrations of 1-10 microM as part of a continuum of injury, which at its most servere was purely necrotic. Cells, maintained in chemically defined neurobasal/B27 medium, were exposed at d7 for 2 h to Glu (1-500 microM), and cellular injury was analysed 2 and 24 h after insult using morphology (phase-contrast microscopy), a 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay, nuclear staining with 4,6-diamidino-2-phenylindole (DAPI), terminal transferase-mediated dUTP nick end labelling (TUNEL) and DNA fragmentation by gel electrophoresis. Glu-mediated neurotoxicity was prevented by MK-801 (5 microM), whilst CNQX (20 microM) attenuated injury by 20%. Exposure to intensive insults (100 and 500 microM Glu) induced necrosis characterized by rapid cell swelling (< 2 h) and lack of chromatin condensation, confirmed by DAPI nuclear staining. In contrast, mild insults (< 20 microM Glu) failed to produce acute neuronal swelling at < 2 h, but 24 h after injury resulted in a large number of apoptotic nuclei as confirmed by TUNEL and electrophoretic evidence of DNA fragmentation, which was attenuated by cycloheximide (0.1 microg/ml). Our findings indicate for the first time that physiological concentrations of Glu produce neuronal injury across a continuum involving apoptosis (< 20 microM) and increasingly necrosis(> 20 microM), dependent on the severity of the initial insult. PMID- 9849678 TI - Dissociation between the Joro spider toxin sensitivity of recombinant alpha-amino 3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and their ability to increase intracellular calcium. AB - We compared the toxin sensitivity, Ca2+ flux response and rectification properties of recombinant alpha-amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (AMPA) receptors obtained by transfecting human embryonic kidney (HEK) 293 cells with different ratios of GluR1 and GluR2 cDNAs (10:1 to 1:10). Simultaneous measurements of kainate-activated Ca2+ fluxes and inward currents, using fura-2 microfluorimetry under voltage clamp conditions, suggested the existence of GluR2 containing channels which are permeable to Ca2+ and insensitive to Joro spider toxin (JSTx). Imaging experiments showed that JSTx inhibition of the Ca2+ response induced by kainate was reduced by increasing the relative amount of GluR2. However, even at GluR1/GluR2(R) ratios of 1:1 and 1:4, cells were still able to flux Ca2+ when stimulated by kainate. GluR2 similarly inhibited the ability of JSTx to reduce kainate-evoked inward currents in whole cell patch clamp experiments. Variations in the rectification properties of the AMPA currents, induced by changes in the cDNA ratio, were not always correlated with the changes in toxin sensitivity and [Ca2+]i response. Thus, cells with almost linear I-V relationships were partially blocked by JSTx and still Ca2+ permeable. Our results indicate a dissociation between the toxin sensitivity and Ca2+ flux through GluR2 containing AMPA receptors and suggest that receptors with diverse Ca2+ permeabilities are generated by the expression of variable amounts of GluR2. PMID- 9849679 TI - Progressive hearing loss, hypoplasia of the cochlea and widened vestibular aqueducts are very common features in Pendred's syndrome. AB - Long-term hearing threshold-on-age follow-up data, including non-linear regression analysis, are given for 12 consecutive Pendred patients. The clinical diagnosis of Pendred's syndrome was confirmed by a mutation analysis of the PDS gene in 11 out of the 11 cases tested. Recent imaging of the temporal bones in seven out of these 12 patients showed widened vestibular aqueducts in each case. The diagnostic perchlorate test was negative in one patient, but this test was positive in her affected sister. Mutation analysis of the PDS gene in these patients confirmed that Pendred's syndrome is a monogenetic disorder. Progressive sensorineural hearing loss and widened vestibular aqueducts are characteristic features of Pendred's syndrome, which provides the opportunity to diagnose Pendred's syndrome clinically in the first few years of life, as has recently been suggested in a case report (Cremers et al., Progressive sensorineural hearing loss and a widend vestibular aqueduct in Pendred syndrome, Arch. Otolaryngol. 124 (1998) 501-505). Mutation analysis of the involved gene can be used to confirm the clinical diagnosis. PMID- 9849680 TI - Relationship between chronic nasal obstruction and craniofacial growth: an experimental model. AB - The aim of this paper was to verify if the growth of the nasomaxillary complex can be influenced by a purely functional alteration such as nasal obstruction, which was induced experimentally in a genetically controlled animal model. Sixty albino rats were employed. Twenty of them had the right nostril occluded by a synthetic resin; another twenty had both nostrils occluded; the other 20 were taken as control group. When the growth was completed, the rats were sacrificed and cephalometric analysis was carried out. Both treated groups showed a statistically significant reduction in overall weight and height, in the vertical development of the nasomaxillary complex and in the skullbase longitudinal axis. After discussing the literature on the subject, the authors conclude that normal craniofacial growth in the rat must somehow depend on physiological nasal breathing, which should therefore be considered of crucial importance. PMID- 9849681 TI - The velo-cardio-facial syndrome: the otorhinolaryngeal manifestations and implications. AB - The velo-cardio-facial syndrome (VCFS), due to a deletion in chromosome 22 on its long arm (22q11), is a leading cause of velopharyngeal dysfunction and cleft palate. With the recent finding of a deletion on chromosome 22q11 in these patients with velopharyngeal dysfunction, a routine test is available making the diagnosis of VCFS much more frequent than previously thought. PMID- 9849682 TI - Daily tympanometry in children during the cold season: association of otitis media with upper respiratory tract infections. AB - The causal association between otitis media and viral upper respiratory tract infections (URI) suggests that early intervention during the course of a viral URI could prevent many episodes of otitis media. However, the feasibility of this approach can not be assessed at present since many aspects of the epidemiology and natural history of URI-associated otitis media are undefined. To address this deficiency, daily monitoring of middle ear status (tympanometry) and cold symptoms and weekly pneumatic otoscopy were done on a pilot cohort of 20 children. These children, between the ages of 2 and 6, were followed from November 1996 to April 1997. Compliance with symptom diaries was 85%, with tympanograms was 90%, and with weekly physician visits was 70%. During the study period, there were 53 'colds' (average 2.65 per child) and 28 new episodes of middle ear effusion (10 unilateral and 9 bilateral). Overall, 47.3% of the tympanograms were Type A, 17.2% Type C1, 9.4% Type C2, and 21.8% Type B. Children who developed MEE during the study spent more time with abnormal MEP (either MEP < -150 or flat) during both cold and healthy days than children who did not develop MEE. Temporally, during colds, high negative pressures preceded the development of Type B tympanograms in children who developed middle ear effusions. Children who did not develop effusions still had high negative pressures during colds, but recovered to a normal pressure within days. These observations document the feasibility of this investigational format for study of the sequential changes in middle ear status before, during and after a URI. PMID- 9849683 TI - Temporal bone CT findings in the CHARGE association. AB - Ten out of 20 cases with the CHARGE association and two CHARGE-like cases underwent temporal bone CT scanning and/or MRI: they all showed bilateral aplasia of the semicircular canals and obliteration of the oval windows. Vestibular examination was performed in nine CHARGE cases and the two CHARGE-like cases, which disclosed vestibular areflexia in all of them. Of the 16 evaluable CHARGE cases, eight had bilateral mixed hearing impairment, while eight had sensorineural hearing impairment which was bilateral in six and unilateral in two cases. Temporal bone CT scanning is therefore indicated in suspected CHARGE cases, even if they show normal hearing or a relatively good bone conduction threshold in one or both ears. PMID- 9849684 TI - The silent epidural abscess as a complication of acute otitis media in children. AB - Acute otitis media with complications is a persistent problem in the modern antibiotic era with a relatively high mortality rate. Acute mastoiditis is a serious complication of acute otitis media with potentially grave consequences and the epidural abscess constitutes the commonest of all intracranial complications, arising from middle ear infections. We report two cases of children with acute mastoiditis without evidence of intracranial complication or subperiosteal abscess, in whom the early evaluation with computed tomography (CT) disclosed an unsuspected epidural abscess. Therefore, we stress the use of CT as a rule of thumb for every child with acute mastoiditis. PMID- 9849685 TI - Sudden total bilateral deafness due to asymptomatic mumps infection. AB - Mumps is the most common cause of unilateral acquired sensorineural hearing loss in children. Although it usually affects the salivary glands. the inner ear may be involved. Deafness is usually unilateral, sudden in onset, profound and permanent. Bilateral total sensorineural hearing loss had been rarely reported in English literature. We present a case of total deafness due to asymptomatic mumps infection. PMID- 9849686 TI - The near complete tracheal ring deformity. AB - Congenital complete and near-complete tracheal rings are extremely rare tracheal deformities. There are fewer than 100 cases of congenital complete tracheal rings, and only four cases of near-complete described in the literature. Long segment occurrences of these anomalies are usually incompatible with life. We describe the presentation of the long-segment near complete tracheal ring deformity and long-segment anterior fusion of cartilage rings (cartilage sleeve deformity) in a living infant. The literature is reviewed, and the embryology, presentation, and management of these extremely rare anomalies are discussed. PMID- 9849688 TI - Extradural frontal abscess complicating nasal septal abscess in a child. AB - Nasal septal abscess (NSA) is an uncommon sequel to minor nasal trauma. Abscess extension beyond the nasal cavity is rarely documented. A case of a 10-year-old boy who presented with a NSA associated with a large extradural frontal abscess is presented and indications for CT scanning in the workup of pediatric patients with NSA is discussed. PMID- 9849687 TI - Pharyngo-laryngeal histoplasmosis: one case in an immunocompetent child. AB - We report a very rare case of disseminated pharyngo-laryngeal histoplasmosis with systemic spread in a 10-year-old, immunocompetent child from Guyana. The main signs were a marked deterioration in his general condition, hepato-splenomegaly, multiple lymphadenopathy and ulcerated pharyngo-laryngeal lesions. The diagnosis was made from brushings of the ulcerative lesions, a lymph node biopsy and serological tests performed in the national reference center for histoplasmosis. The initial treatment was with amphotericin B, which was then replaced by oral itraconazole. We report here the main epidemiological, clinical and therapeutic characteristics. PMID- 9849689 TI - Stem cell mobilisation in lymphoproliferative diseases. AB - A number of different regimens have evolved for the mobilisation of peripheral blood stem cells for autologous transplantation in patients with lymphoma or myeloma. A successful regimen could be defined as one which consistently resulted in the collection of an optimal number of CD34+ cells with a minimum number of apheresis procedures with minimal toxicity. Initial protocols, which used chemotherapy alone as a mobilising agent, have now been replaced by regimens involving the use of haematopoietic growth factors either alone or in combination with variable doses of cyclophosphamide. Although there is good evidence that high-dose cyclophosphamide (6-7 g/m2) is an effective mobilising agent it is associated with significant toxicity and many groups have now utilised lower doses of cyclophosphamide with reduced toxicity which have still proven to be effective in the majority of patients. More recently a number of 'second generation' combined salvage chemotherapy and mobilisation regimens have been reported for use in the lymphomas which have the advantage of avoiding a specific stem cell mobilisation step and at the same time appear more consistently effective at mobilising stem cells than cyclophosphamide and G-CSF. These regimens are associated with fewer 'poor-mobilisers' and indeed some patients who have failed previous mobilisation with cyclophosphamide and G-CSF have been successfully re-mobilised. It is clear that in both lymphoma and myeloma patients the success of PBSC mobilisation is affected by the amount and type of previous chemotherapy and radiotherapy and probably other pre-treatment factors as exemplified by variability seen in normal donors mobilised with G-CSF alone. In myeloma most groups have utilised cyclophosphamide in variable doses in combination with G-CSF or GM-CSF. However, recent randomised studies have confirmed that G-CSF alone is an effective and nontoxic alternative although it appears that the efficacy of G-CSF as a single agent is related to the dosage used with daily doses of 16 microg/kg/day or greater being most effective. Thus, disease-specific mobilisation strategies appear to be emerging and these will undoubtedly be modified further as more is understood concerning the biology of blood stem cell mobilisation. PMID- 9849690 TI - Hematopoietic stem cell transplantation for infantile osteopetrosis. AB - Infantile osteopetrosis is a lethal disorder resulting from a severe defect in the ability of osteoclasts to resorb bone. The only therapy shown to be capable of providing lasting benefit is allogeneic hematopoietic stem cell transplantation (HCT). We report the outcome of 10 patients with infantile malignant osteopetrosis treated with HCT from an HLA A, B, DRB1 matched (n=6) or A or B locus mismatched (n=4) family member or unrelated donor at the University of Minnesota between 1978 and 1997. Eight of 10 patients achieved primary engraftment; secondary graft failure was seen in two patients. Five of 10 patients survive; three with full or partial donor chimerism and two with autologous hematological recovery. Transient or partial donor chimerism can be sufficient to correct the hematological manifestations of osteopetrosis. We recommend early referral for consideration of HCT with a related or unrelated donor as neurosensory manifestations of osteopetrosis are generally not reversible. Donor engraftment may be easier to achieve early in the course of the disease. PMID- 9849691 TI - CD34+ stem cell augmentation of elutriated allogeneic bone marrow grafts: results of a phase II clinical trial of engraftment and graft-versus-host disease prophylaxis in high-risk hematologic malignancies. AB - Although T cell depletion of allografts used in BMT has reduced GVHD, it has been associated with inferior engraftment and an increased risk of relapse. We have found that T cell depletion by counterflow centrifugal elutriation (CCE) also results in depletion of CD34+ stem cells. In order to determine if the discarded CD34+ cells would improve engraftment, we undertook a phase II trial of allogeneic BMT in which 110 patients (median age 43) with a variety of hematologic malignancies received CD34+ stem cell augmented, elutriated marrow grafts. The T cell-depleted grafts were tightly controlled and contained a mean of 4.3 x 10(7) mononuclear cells/kg, 3.3 x 10(6) CD34+ cells/kg, 1.5 x 10(5) CFU GM/kg and 5.5 x 10(5) CD3+ T cells/kg. Median time to engraftment of granulocytes (>500/microl) was 16 days and of platelets (>50000/microl) was 25 days, comparable to that seen with unmanipulated marrow. No mixed hematopoietic chimerism was observed that was not associated with disease relapse. The four patients (3.6%) who failed to engraft were all at high risk because of prior donor transfusions or underlying marrow disorders. The incidence of GVHD was dependent on the duration of cyclosporin A (CsA) immunosuppression. In patients who received CsA for > or = 80 days, the incidence of clinically significant acute GVHD (>stage 1) and extensive, chronic GVHD was 5% and 11%, respectively. Peritransplant (< or = 100 day post-BMT) mortality for this group of patients was 15%. Event-free survival in selected subsets of patients compared favorably to previous studies in which patients received unmanipulated marrow allografts. PMID- 9849692 TI - Autologous transplantation of mobilized peripheral blood CD34+ cells selected by immunomagnetic procedures in patients with multiple myeloma. AB - In the use of autologous PBPC transplantation in patients with multiple myeloma, contamination of PBPC with myeloma cells is commonly observed. Enrichment for CD34+ cells has been employed as a method of reducing this contamination. In this study the reduction of myeloma cells in PBPC was accomplished by the positive selection of CD34+ cells using immunomagnetic bead separation (Isolex 300 system). PBPC were mobilized from 18 patients using cyclophosphamide (4.5 g/m2) and G-CSF (10 microg/kg/day). A median of two leukaphereses and one selection was performed per patient. The median number of mononuclear cells processed was 3.50 x 10(10) with a recovery of 1.11 x 10(8) cells after selection. The median recovery of CD34+ cells was 48% (range 17-78) and purity was 90% (29-99). The median log depletion of CD19+ cells was 3.0. IgH rearrangement, assessed by PCR, was undetectable in 13 of 24 evaluable CD34+ enriched products. Patients received 200 mg/m2 of melphalan followed by the infusion of a median of 2.91 x 10(6)/kg CD34+ cells (1.00-16.30). The median time to absolute neutrophil count >0.5 x 10(9)/l was 11 days, and sustained platelet recovery of >20 x 10(9)/l was 14 days. We conclude that immunomagnetic-based enrichment of CD34+ cells results in a marked reduction in myeloma cells without affecting engraftment kinetics. PMID- 9849693 TI - A randomized trial of granulocyte colony-stimulating factor (Neupogen) starting day 1 vs day 7 post-autologous stem cell transplantation. AB - The purpose of the study was to evaluate the effect of delayed granulocyte colony stimulating factor (G-CSF) use on hematopoietic recovery post-autologous peripheral blood progenitor cell (PBPC) transplantation. Patients were randomized to begin G-CSF on day +1 or day +7 post transplantation. Thirty-seven patients with lymphoma or myeloma undergoing high-dose therapy and autologous PBPC rescue were randomized to daily subcutaneous G-CSF beginning on day +1 or day +7 post transplant. Patients < or =70 kg received 300 microg/day and >70 kg 480 microg/day. All patients were reinfused with PBPCs with a CD34+ cell count >2.0 x 10(6)/kg. Baseline characteristics of age, sex and CD34+ cell count were similar between the two arms, the median CD34+ cell count being 5.87 x 10(6)/kg in the day +1 group and 7.70 x 10(6)/kg in the day +7 group (P=0.7). The median time to reach a neutrophil count of >0.5 x 10(9)/l was 9 days in the day +1 arm and 10 days in the day +7 arm, a difference which was not statistically significant (P=0.68). Similarly, there was no difference in median days to platelet recovery >20000 x 10(9)/l, which was 10 days in the day +1 arm and 11 days in the day +7 arm (P=0.83). There was also no significant difference in the median duration of febrile neutropenia (4 vs 6 days; P=0.7), intravenous antibiotic use (7 vs 8 days; P=0.54) or median number of red blood cell transfusions (4 vs 7 units; P=0.82) between the two arms. Median length of hospital stay was 11 days post PBPC reinfusion in both groups. The median number of G-CSF injections used was 8 in the day +1 group and 3 in the day +7 group (P < 0.0001). There is no significant difference in time to neutrophil or platelet recovery when G-CSF is initiated on day +7 compared to day +1 post-autologous PBPC transplantation. There is also no difference in number of febrile neutropenic or antibiotic days, number of red blood cell transfusions or length of hospital stay. The number of doses of G-CSF used per transplant is significantly reduced with delayed initiation, resulting in a significant reduction in drug costs. For patients with an adequately mobilized PBPC graft, the initiation of G-CSF can be delayed until day +7 post-PBPC reinfusion. PMID- 9849694 TI - Frequent detection of tumor cells in hematopoietic grafts in neuroblastoma and Ewing's sarcoma. AB - Many poor-risk neuroblastomas and tumours of the Ewing's sarcoma family (ET) recur despite autologous transplants. Recurrence may be due to tumor cells contained in the BM harvests or PBSC harvests. The objectives of this prospective study were to: (1) determine the incidence and degree of tumor cell contamination in paired BM and PBSC harvests; and (2) determine the efficacy of tumor cell purging by immunomagnetic CD34+ cell selection. 198 samples from 11 consecutive patients with neuroblastoma or Ewing's sarcoma were analyzed. We assayed tumor contamination by RT-PCR assay for PGP 9.5, plus immunohistochemistry for neuroblastoma-specific antigens (the latter in neuroblastoma only). None of these patients had tumor cells detected in their BM by clinical histology immediately before BM or PBSC harvests. However, 82% of PBSC and 89% of backup BM harvests were contaminated with tumor by RT-PCR and/or immunocytochemistry assays. Unselected PBSC and BM harvests contained similar quantities of tumor cells (median, approximately 200000 cells). Cyclophosphamide plus G-CSF mobilization did not affect the incidence or level of contamination in PBSC harvests, as compared to blood obtained before mobilization. Immunomagnetic CD34+ cell selection depleted tumor cells by a median of 3.0 logs for PBSC, and 2.6 logs for BM harvests. PMID- 9849696 TI - Ovarian function after autologous bone marrow transplantation in childhood: high dose busulfan is a major cause of ovarian failure. AB - We studied pubertal status and ovarian function in 21 girls aged 11-21 years who had earlier received 1.2-13 years (median 7 years) high-dose chemotherapy and autologous BMT without TBI for malignant tumors. Ten of them were given busulfan (600 mg/m2) and melphalan (140 mg/m2) with or without cyclophosphamide (3.6 g/m2). Eleven others did not receive busulfan. Twelve girls (57%) had clinical and hormonal evidence of ovarian failure. Among nine others who had completed normal puberty, six had normal gonadotropin levels, one had elevated gonadotropin levels and two had gonadotropin levels at the upper limit of normal. The 10 girls who received busulfan all developed severe and persistent ovarian failure. High dose busulfan is therefore a major cause of ovarian failure even when given in the prepubertal period. These findings emphasize the need for long-term endocrine follow-up of these patients in order to initiate estrogen replacement therapy. PMID- 9849695 TI - Lymphoproliferative disorders following allogeneic bone marrow transplantation: the Vancouver experience. AB - Between June 1988 and May 1996, 428 patients underwent allogeneic BMT (288 related donor (RD) and 140 unrelated donor (UD)) at the Vancouver General Hospital. Eight patients (UD six and RD two) developed a post-transplant lymphoproliferative disorder (PTLD). Median age at BMT was 38 years (range 22 51). Five of the six UD allografts were T cell depleted. Cyclosporine+/ methotrexate was used for GVHD prophylaxis. All eight patients developed GVHD; in six this was refractory to treatment with corticosteroids. Rabbit antithymocyte globulin (ATG) or an anti-CD5-ricin A chain immunotoxin (Xomazyme) was used as second-line therapy for GVHD. Presentation with PTLD occurred at median day 90.5 (range 34-282) post BMT. Five of the eight patients had a rapidly progressive course characterized by fever, lymphadenopathy, lung and liver involvement and died within 3-8 days. PTLD was an incidental finding at post mortem examination in two patients. The remaining patient had localized disease and recovered. Pathological analysis revealed two morphological patterns; diffuse large B cell lymphoma (DLBC lymphoma, five patients) and polymorphous B cell hyperplasia (PBCH, three patients). EBV expression was positive in all eight cases and monoclonality was demonstrated in seven cases. In multivariate analysis, T cell depletion of the allograft (P=0.0001, relative risk (RR)=30.5), anti-T cell therapy for GVHD (P=0.006, RR=12.7) and acute GVHD grades 3-4 (P=0.04, RR=7.7) were the significant factors for development of PTLD. In conclusion, we have identified two forms of PTLD after BMT: one is characterized by disseminated disease with a rapidly progressive and often fulminant course and the other by localized, relatively indolent disease. Morphology, EBV positivity and clonality do not appear to correlate with the clinical course. The major risk factors for development of PTLD after BMT are ex vivo T cell depletion of the allograft and in vivo anti-T cell therapy for GVHD. PMID- 9849697 TI - Late-onset hemorrhagic cystitis after hematopoietic stem cell transplantation in children. AB - We analyzed the incidence, complications, and risk factors for late-onset hemorrhagic cystitis (HC) in 256 children undergoing hematopoietic stem cell transplantation (HSCT). Twenty-six recipients (10.2%) developed late-onset HC between 3 and 270 days (median, 33 days) after HSCT. In most patients, the severity of HC was mild to moderate, and spontaneous resolution occurred. Three children developed bladder tamponade, and one required suprapubic cystotomy. Four children died in the early post-transplant period without resolution of HC, but HC was not the direct cause of death in any patient. Twenty-two patients recovered within 6-86 days (median, 16 days) of onset. Three predisposing factors were identified for development of late-onset HC by multivariate analysis: allogeneic HSCT, older age (> or = 7 years), and busulphan for pretransplant conditioning were significantly associated with late-onset HC (P=0.022, P=0.044 and P=0.036, respectively). Excretion of adenovirus type 11 was demonstrated in six of 22 patients at the onset of cystitis. We suspect that reactivation of virus may be a major pathogenic factor in late-onset HC, but several clinical factors are also associated. PMID- 9849698 TI - Increased platelet transfusion requirement is associated with multiple organ dysfunctions in patients undergoing hematopoietic stem cell transplantation. AB - Organ dysfunction following hematopoietic stem cell transplantation (HSCT) may be a manifestation of a systemic inflammatory response. We speculate that part of the platelet transfusion requirement in HSCT patients results from this systemic inflammatory response, and increased transfusion requirement would be associated with, or precede, organ dysfunction. We studied 199 adults undergoing autologous (n=173) or allogeneic (n=26) HSCT. Patients with CNS (P=0.008) or pulmonary (P=0.002) dysfunction, or with VOD (P=0.05) received a higher mean number of platelet transfusions per week than patients who did not have these dysfunctions. Furthermore, a higher number of platelet transfusions during any 1 week period was significantly associated with development of pulmonary (P=0.0002) or renal (P < 0.0001) dysfunction in the following week. This predictive value was strongest early in the HSCT course, but remained significant over all 4 weeks. In multivariate analysis the number of platelet transfusions during the previous week was independently predictive for development of pulmonary dysfunction in week 2 (P=0.01) and week 3 (P=0.055). We believe that occurrence of increased platelet transfusion requirement prior to onset of dysfunction is consistent with the concept that an antecedent inflammatory response results in both platelet consumption and various organ dysfunctions. Increased platelet transfusion requirement may act as an early marker of subsequent organ dysfunction. Additionally, there may be a direct role of platelets in the development and progression of organ dysfunction in HSCT patients. PMID- 9849699 TI - IFN-gamma-mediated prevention of graft-versus-host disease: pharmacodynamic studies and influence on proliferative capacity of chimeric spleen cells. AB - Recently we demonstrated that prolonged administration of IFN-gamma prevented the development of GVHD in a MHC-mismatched murine BMT model. Treatment with IFN gamma allowed the development of mature donor-derived allo-tolerant immunocompetent cells in complete chimeric recipients. Here we present data on the pharmacodynamics of this cytokine-mediated protection against GVHD. Treatment with 50000 U IFN-gamma twice weekly for a period of 5 weeks, starting at the day of BMT, was shown to be the optimal treatment protocol, resulting in complete prevention of GVHD-related mortality. Treatment during 1 week with a three-fold higher weekly dose of IFN-gamma (50000 U six times) did not result in significantly improved survival. The start of IFN-gamma administration was a critical factor since a delay of 3 days from the time of BMT resulted in substantial GVHD-induced mortality. Furthermore, it was shown that IFN-gamma treatment inhibited the spontaneous and Con-A-induced proliferation of T cells at 7-14 days after BMT, which is the critical period for the initiation of acute GVHD. However, long-term survivors after IFN-gamma treatment showed a recovery of immunity in contrast to long-term survivors of saline-injected animals, as tested by Con-A responsiveness. It seems that injection of high dose IFN-gamma suppresses the response of potentially alloreactive donor T cells during what normally is the initiation phase of the GVH reaction (GVHR), resulting in the abrogation of GVHD. PMID- 9849700 TI - Clinically applicable bulk isolation of blood CD34+ cells for autografting in children. AB - CD34+ cells were purified in bulk from apheresis-collected cells of children with cancer using monoclonal antibody (MoAb) and magnetic beads (Baxter ISOLEX system). To improve the purity of the final product for possibly better tumor cell purging and to make the manufacturer's original procedure more cost effective, we incubated the cells for 30 min with l-phenylalanine methylester hydrochloride (PME) to reduce the cell number by removing contaminating granulocytes and monocytes in the initial step before incubation with MoAb. Our modification prevented nonspecific interactions between MoAb and magnetic beads, and thereby saved expensive materials for purification. A total of 40 purifications were performed with samples containing a mean of 3.1 x 10(9) blood cells mobilized from 15 children by chemotherapy plus granulocyte colony stimulating factor (G-CSF). The entire purification procedure, from the end of apheresis to storage, was completed within 5h. After incubation with PME and double-layered (40/60%) Percoll separation, the number of CD34+ cells was reduced to 48+/-29%, which suggests the possibility that half of the CD34+ cells in the inoculum were nonclonogenic in the hematopoietic progenitor assay. PME/Percoll treated cells were then subjected to a final isolation procedure with MoAb according to the manufacturer's suggestions, and 52+/-42% and 32+/-22%, respectively, of the CFU-GM and CD34+ cells present in the initial bag inoculums were recovered. The recovery rates were, respectively, 54% and 67%, when the calculation was limited to the isolation procedure with MNoAb. The purity of isolated CD34+ cells and the plating efficiency in methylcellulose culture were, respectively, 77+/-24% and 33+/-13%. Fourteen children were subsequently autografted with purified CD34+ cells after marrow ablative chemotherapy. The median number of days to achieve an ANC of 0.5 x 10(9)/l was 12 and that to achieve a platelet count of 50 x 10(9)/l was 22.5, which were comparable to those in our historical group of 55 patients who underwent transplant with unmanipulated blood cells (13 and 16 days). These results suggest that our modified purification procedure with PME is useful for the initial reduction of cell numbers to save costly materials, and that cells isolated by this procedure can be directly used in clinical transplantation procedures. PMID- 9849701 TI - Complete response in severe thrombotic microangiopathy post bone marrow transplantation (BMT-TM) after multiple plasmaphereses. AB - A 44-year-old male with Ph+ chronic myeloid leukaemia (CML) underwent histoidentical allogeneic bone marrow transplantation 18 months after initial diagnosis. He received pretransplant conditioning with busulphan and cyclophosphamide (Bucy). GVHD prophylaxis consisted of methotrexate, cyclosporine (CsA) and methylprednisolone. On day +50, he developed a microangiopathic haemolytic anaemia with indirect bilirubinaemia, 10% fragmented red cells (FC) and an elevated LDH (1213 U/l: normal range 100-185 U/l). Clinical symptoms consisted of edema and hypertension. The patient was not febrile and had no neurological changes. A clinical diagnosis of severe (grade 4) multifactorial (acute GVHD, CMV infection and cyclosporine) BMT-TM was made. He responded following 19 plasma exchanges with replacement with fresh frozen plasma. PMID- 9849702 TI - Fournier's gangrene: a clinical presentation of necrotizing fasciitis after bone marrow transplantation. AB - Three patients with ANLL developed Fournier's gangrene as an early complication after allo-BMT (two cases) and auto-BMT (one case); two patients were in first CR, the third had resistant disease. Patients developed fever, perineal pain, swelling and blistering of the genital area. Pseudomonas aeruginosa was isolated from the lesions and patients received systemic antibiotic therapy, surgical debridement and medication with potassium permanganate solution. Two patients made a complete recovery although one died of sepsis. The third had progressive involvement of the abdominal wall and later died of leukemia. Early diagnosis of this disorder and prompt initiation of appropriate therapy can prevent progression of this acute necrotizing infection. PMID- 9849703 TI - G-CSF dosage and scheduling following autologous PBSC transplantation. PMID- 9849704 TI - Thromboxane and prostacyclin in maternal and fetal circulation in pre-eclampsia. AB - OBJECTIVES: A major pathophysiologic change of pre-eclampsia has been attributed to the overproduction of thromboxane A2 (TXA2) mainly from activated platelets. On the other hand, increased biosynthesis of TXA2 has also been reported from preeclamptic placentas. The systemic role of these different sources of TXA2 has not been clarified. The purpose of this study is to define the changes of TXA2 and the antagonizing prostacyclin (PC) in maternal and fetal circulations. METHODS: The stable metabolites of TXA2 and PC [Thromboxine B2 (TXB2) and 6-keto prostaglandin F1alpha (6-keto-PGF1alpha), respectively] in the cord and maternal blood of nine patients with pre-eclampsia and nine normal parturients were measured by radioimmunoassay. RESULT: In normal pregnancy, the cord blood contained much higher TXB2 (1697+/-898 vs. 267+/-128 ng/ml, P < 0.01) and 6-keto PGF1alpha (266+/-263 vs. 12.5+/-3.9 ng/ml, P < 0.05) levels than the maternal blood. In the preeclamptic state, a marked increase of TXB2 was noted in both maternal and cord blood, reaching levels which were significantly higher than during normal pregnancy (2995+/-1103 vs. 267+/-128 ng/ml in maternal blood, P < 0.0001, and 3197+/-1288 vs. 1697+/-898 ng/ml in cord blood, P < 0.005). A less significant increase in 6-keto-PGF1alpha (134+/-10.8 vs. 12.5+/-3.9 ng/ml, P < 0.05) was also noted in the maternal blood. Moreover, the level of TXB2 correlated with the diastolic blood pressure of preeclamptic patients before and after delivery. CONCLUSION: The results suggest an abundant source of eicosanoids in the feto-placental circulation, which does not readily cross the placental barrier. In pregnancy complicated with pre-eclampsia, thromboxane level of both fetal and maternal circulations are markedly increased, which may be responsible for the pathophysiologic changes. The lack of adverse systemic effects on the fetus highlights a placental source of TXA2 of transient bioactivity which is rapidly hydrolyzed to non-active TXB2. Federation of Gynecology and Obstetrics PMID- 9849705 TI - Obstetric outcome of elderly low-risk nulliparae. AB - OBJECTIVE: To compare referrals and reasons for referral during pregnancy and labor, mode of delivery and obstetric outcome of first births in women 35 years and older with women 20-30 years old. METHODS: A prospective cohort study was performed of 146 elderly and 306 younger nulliparae in seven independent midwives' practices in and around Amsterdam. RESULTS: No significant differences in referrals were found between the two compared groups. After selection during pregnancy, obstetric outcome was not different between the groups. A higher percentage of episiotomies was found in the elderly group, compared to the younger group. CONCLUSIONS: After proper selection during pregnancy, the elderly nullipara under the care of a midwife does not have an increased risk of fetal distress or other emergency factors, compared to the younger nullipara. However, high referral rates during labor - both of younger and older women - were observed in this study. PMID- 9849706 TI - Fracture of the clavicle in the newborn following normal labor and delivery. AB - OBJECTIVE: Earlier works have associated neonatal clavicular fracture (0.2-3.5% of all deliveries) with a range of procedural, fetal and maternal risk factors; more recent studies, however, have failed to firmly identify any one or a combination of them. In the present work we sought to determine possible ante/intra-partum risk factors for this condition. STUDY DESIGN: Using a retrospective case-controlled approach, we examined a series of maternal, fetal and pregnancy or delivery-related variables in 87 cases (out of 403) of fractured clavicle of the newborn diagnosed in our department from 1986 to 1994. All infants were delivered vaginally (in the occipito-anterior position) at term by a specialist obstetrician and underwent peripartum sonographic fetal weight estimation. All variables were compared with those of an equal number of infants born immediately before or after each affected infant and delivered by the same obstetrical team. RESULTS: Fractured clavicles were found in 1.65% of the total number of deliveries during the study period. Neonatal clavicular fracture was significantly and directly related to the duration of the second stage of labor, peripartum sonographic fetal weight estimation, and neonatal length, and inversely related to maternal height; nevertheless, all values in both the study and control groups were within normal range. Logistic regression analysis showed that these antenatal variables significantly affect the chances of clavicular fracture, but due to the high false-positive rate they cannot serve clinically as a comprehensive antenatal prediction index. CONCLUSIONS: The majority of clavicular fractures occur in normal newborns following normal labor and delivery. The risk factors we identified statistically do not offer a method for clinical prenatal prediction. This work provides statistical evidence of the nature of this complication of early newborn life. PMID- 9849707 TI - Reproducibility and diagnostic sensitivity of ultrasonometry of the phalanges to assess osteoporosis. AB - OBJECTIVE: The present study was designed to assess the reproducibility and the diagnostic sensitivity of the amplitude-dependent speed of sound (SoS) at the distal metaphysis of the proximal phalanges. METHOD: Fourteen presumably healthy volunteers were repeatedly measured every 6 weeks for approximately 6 months in order to assess the reproducibility of the SoS of the phalanges. We recruited 91 post-menopausal women, aged 55-75 years, who were divided in three groups according to their lumbar bone mineral density (BMD) and the existence of prevalent vertebral fractures. The objective was to evaluate the diagnostic sensitivity of SoS measurements. We used DBM Sonic 1200 equipment, and assessed the velocity at which US cross the phalanx in a lateral-medial direction. In post menopausal women, BMD was measured by dual energy X-ray absorptiometry (DXA) at the level of the lumbar spine, the total zone of the non-dominant hip and the femoral neck zone of the non-dominant hip. RESULTS: The precision of the SoS measurements was 0.71+/-0.05% (mean+/-S.E.M) whereas the reproducibility was 0.95+/-0.06%. Subjects with low BMD or prevalent fractures had significantly lower values of SoS (P < 0.001) than the controls. ROC curve analysis applied to the study population confirmed that SoS was able to discriminate between the controls and osteoporotic subjects (area under the ROC curves were 0.82 (low bone mineral density) and 0.85 (prevalent fractures), respectively). Hip BMD was found to be the most significant variable when comparing the controls and the low density patients by stepwise discrimination and SoS significantly improved the discrimination between the groups when added to the hip BMD. The hip BMD was again the most discriminant variable when applying the same techniques to controls and patients with prevalent fractures, followed by SoS and lumbar BMD. A cut-off value of 1881 m/s is defined for SoS by logistic discrimination and likelihood ratio function. With this value, the sensitivity and the specificity for SoS used in the diagnosis of established osteoporosis were, 81.5% and 79.3%, respectively. Sensitivity and specificity were significantly improved when combining ultrasonometry and densitometry. CONCLUSION: Measurement of ultrasound velocity at the phalanges appears to be a precise and reproducible technique. SoS discriminates between normal post-menopausal women and patients with either low lumbar BMD or prevalent fractures to the same extent as BMD measurements. PMID- 9849708 TI - Prospective randomized comparison between an open and closed vaginal cuff in abdominal hysterectomy. AB - OBJECTIVE: To compare an operative and postoperative course of open vaginal cuff hysterectomy and closed vaginal cuff hysterectomy, and to correlate the length of stay, febrile morbidity and the incidence of pelvic fluid collections to the type of surgery. PARTICIPANTS: One-hundred women scheduled for hysterectomy were prospectively randomized into two groups that underwent either a closed or an open vaginal cuff technique. RESULTS: The open vaginal cuff technique took on average 19% more time than the closed vaginal cuff operation (P < 0.05, t-test). The incidence and size of pelvic fluid collections was significantly higher after the closed vaginal cuff hysterectomy than after the open technique (P < 0.01, t test). However, the postoperative length of stay, febrile morbidity and the rate of complications were similar. CONCLUSIONS: Both techniques of hysterectomy produced a similar postoperative course despite the fact that the closed vaginal cuff technique resulted in a higher incidence of pelvic fluid collections. Therefore considering a shorter operation time for the closed vaginal cuff hysterectomy, this technique seems slightly preferable. PMID- 9849709 TI - Glucocorticoid receptors on mononuclear leukocytes in polycystic ovary syndrome. AB - OBJECTIVE: Many studies have suggested that there is a possible hormonal dysregulation of hypothalamic-pituitary-adrenal (HPA) axis and an increased cortisol clearance in patients with polycystic ovary syndrome (PCOS). Therefore in this study, we have examined the role of glucocorticoid receptor/s (GR) characteristics in the developing of these abnormalities in patients with PCOS. METHOD: For this purpose, the number and affinity of GR in peripheral mononuclear leukocytes (MNL) of 10 patients with PCOS and 10 healthy women (controls) were determined. RESULTS: There were no significant differences in the number (6500+/ 1001 sites/cell and 6352+/-1697 sites/cell, respectively; P > 0.05) and affinity (3.93+/-0.89 nM and 4.49+/-0.71 nM, respectively; P > 0.05) of GR between the PCOS patients and the controls. CONCLUSIONS: These results suggest that the alterations in the HPA axis and in the cortisol metabolism observed in PCOS are not related to GR deficiency. PMID- 9849710 TI - Blood loss with mifepristone--misoprostol abortion: measures from a trial in China, Cuba and India. AB - OBJECTIVE: We compared bleeding patterns following early mifepristone-misoprostol abortion with those following early surgical abortions ( < or = 56 days gestation LMP). We analyzed women's reports of their bleeding and clinical measures of blood loss, and modeled the relationships between different bleeding measures and indicators of success and satisfaction. METHOD: Using data from a multicenter trial held in China, Cuba and India, we fit bivariate and multivariate models to analyze bleeding patterns, controlling for a range of demographic, experiential and medical factors (n = 1373). RESULT: Medical abortion patients perceived their bleeding to be heavier than did the surgical patients; however, there were few statistically significant differences in clinical measures of blood loss between the two groups, and blood loss rarely represented a medical problem. Nationality, far more than actual levels of bleeding, influenced women's impressions about blood loss. Although medical abortion patients saw their bleeding as heavier than did the surgical patients, their perceptions of heavier bleeding did not prevent them from having higher satisfaction levels. One exception was that if bleeding exceeded prior expectations, women with the medical method were more likely to be dissatisfied. Pain and cramps, which often accompanied bleeding, were an independently significant factor in predicting satisfaction with the method. CONCLUSION: In order for women to know what to expect with medical abortion, they must be informed beforehand about the level of bleeding that generally accompanies the procedure vs. excessive bleeding that would signal a clinical problem. PMID- 9849711 TI - Extra-amniotic balloon with PGE2 versus extra-ovular Foley catheter with PGF2alpha in mid-trimester pregnancy termination. AB - OBJECTIVE: The objective of this study was to compare the efficacy and side effects of two methods of mid-trimester labor induction, extra-amniotic balloon with intracervical prostaglandin (PG) E2 (0.5 mg x 2) vs. extra-ovular Foley catheter with intrauterine PGF2alpha (1 mg/h x6). METHODS: A cohort of 32 and 36 cases indicated for mid-trimester termination was enrolled and managed with extra amniotic balloon and extra-ovular Foley catheter methods, respectively. Outcomes of induction-to-delivery interval, induction failure, and occurrence of side effects were assessed. RESULTS: There were no statistical differences in maternal age, parity, gestational age or fetal birth weight between the two groups. Compared with the extra-ovular Foley catheter with PGF2alpha group, the induction to-delivery interval was significantly shorter in the extra-amniotic balloon plus PGE2 group. There was no significant difference in side effects and major complications developed in either groups. CONCLUSIONS: The extra-amniotic balloon with intracervical PGE2 is more efficient in reducing the induction-to-delivery interval for termination of mid-trimester pregnancies than the extra-ovular Foley catheter with intrauterine PGF2alpha. PMID- 9849712 TI - Association of fetal gender with pregnancy-induced hypertension and pre eclampsia. PMID- 9849713 TI - Full-term interstitial pregnancy with live birth. PMID- 9849714 TI - Pregnancy outcome among gestational diabetics with blood glucose levels between 7.7 and 8.3 mmol/l. PMID- 9849715 TI - Pulse oxygen saturation and neurologic assessment in human neonates after vaginal and cesarean delivery. PMID- 9849716 TI - Interaction of a nuclear factor derived from the trophoblast cell line BeWo with the HLA-G gene. PMID- 9849718 TI - Placental abruption and intrauterine death following an ant sting. PMID- 9849717 TI - IgG and IgM antibodies of Herpes Simplex Virus type-1 and type-2 in 233 maternal and neonatal sera. PMID- 9849719 TI - Surgico-pathological stage I primary fallopian tube adenocarcinoma. PMID- 9849720 TI - ACOG practice bulletin. Premature rupture of membranes. Clinical management guidelines for obstetrician-gynecologists. Number 1, June 1998. American College of Obstetricians and Gynecologists. AB - The following recommendations are based on good and consistent scientific evidence (Level A): With term PROM, labor may be induced at the time of presentation or patients may be observed for up to 24-72 hours for the onset of spontaneous labor. Antibiotics prolong the latency period and improve perinatal outcome in patients with preterm PROM and should be administered according to one of several published protocols if expectant management is to be pursued prior to 35 weeks of gestation. Antenatal corticosteroids should be administered to gravidas with PROM before 32 weeks of gestation to reduce the risks of respiratory distress syndrome, neonatal intraventricular hemorrhage, necrotizing enterocolitis, and neonatal death. Digital cervical examinations should not be performed in patients with PROM who are not in labor and in whom immediate induction of labor is not planned. Patients with PROM prior to 30-32 weeks of gestation should be managed conservatively if no maternal or fetal contraindications exist. The following recommendations are based primarily on consensus and expert opinion (Level C): Tocolysis may be utilized in patients with preterm PROM to permit administration of antenatal corticosteroids and antibiotics. Antenatal corticosteroids may be administered to gravidas with PROM up to 34 weeks of gestation. PMID- 9849721 TI - ACOG criteria set. Quality evaluation and improvement in practice. Tocolysis. Number 34, June 1998 (replaces No. 31, February 1998). Committee on Quality Assessment. American College of Obstetricians and Gynecologists. PMID- 9849722 TI - Investigation of facial recognition memory and happy and sad facial expression perception: an fMRI study. AB - We investigated facial recognition memory (for previously unfamiliar faces) and facial expression perception with functional magnetic resonance imaging (fMRI). Eight healthy, right-handed volunteers participated. For the facial recognition task, subjects made a decision as to the familiarity of each of 50 faces (25 previously viewed; 25 novel). We detected signal increase in the right middle temporal gyrus and left prefrontal cortex during presentation of familiar faces, and in several brain regions, including bilateral posterior cingulate gyri, bilateral insulae and right middle occipital cortex during presentation of unfamiliar faces. Standard facial expressions of emotion were used as stimuli in two further tasks of facial expression perception. In the first task, subjects were presented with alternating happy and neutral faces; in the second task, subjects were presented with alternating sad and neutral faces. During presentation of happy facial expressions, we detected a signal increase predominantly in the left anterior cingulate gyrus, bilateral posterior cingulate gyri, medial frontal cortex and right supramarginal gyrus, brain regions previously implicated in visuospatial and emotion processing tasks. No brain regions showed increased signal intensity during presentation of sad facial expressions. These results provide evidence for a distinction between the neural correlates of facial recognition memory and perception of facial expression but, whilst highlighting the role of limbic structures in perception of happy facial expressions, do not allow the mapping of a distinct neural substrate for perception of sad facial expressions. PMID- 9849723 TI - Cerebral CT findings in male opioid-dependent patients: stereological, planimetric and linear measurements. AB - Cerebrospinal fluid (CSF) space enlargement has been demonstrated in substance related disorders like alcohol and cocaine dependence. Experimental animal studies showed a reduction in shape and size of mesolimbic dopaminergic neurons after chronic morphine administration. Other studies indicated a change of neurofilament and glial fibrillary acid proteins after chronic opiate administration. Furthermore, frequent overdosing and toxicological effects of 'street'-heroin may lead to CSF space enlargement in opioid dependence. In our study the pericortical and ventricular CSF space of 21 male opioid-dependent patients was compared with an age- and sex-matched normal control group. Considering serious problems with ratio and proportion measures, we used a battery of linear (cella media index, Huckman number, frontal horn index), planimetric (cortical atrophy score) and stereological volumetric measures in order to detect differences in cranial computerized tomography scans. We found a significant ventricular and cortical volume loss of the brain in opioid-dependent patients. A higher degree of frontal lobe volume loss seemed to be associated with a shorter period of abstinence before relapse. However, the etiology of volume loss of the brain in opioid-dependent patients is still unclear, but experimental animal studies provide some evidence that long-term, chronic opiate exposure is associated with visible changes of specific structures in the brain. PMID- 9849724 TI - Effects of high amphetamine dose on mood and cerebral glucose metabolism in normal volunteers using positron emission tomography (PET). AB - The effects of high euphorigenic doses of D-amphetamine (0.9-1.0 mg/kg p.o.) on regional cerebral glucose metabolism (rCMRglu) and psychological measures were investigated in 10 healthy human volunteers using a within-subject design and [F 18]-fluorodeoxyglucose positron emission tomography (FDG-PET) and a variety of psychological assessments. At the dose tested, D-amphetamine produced a mania like syndrome concomitantly with a widespread increase in absolute cerebral metabolism, which was significant in the anterior cingulate cortex, caudate nucleus, putamen, and thalamus. An exploratory analysis revealed that: (1) certain aspects of this mania-like syndrome correlated positively with the metabolic changes seen in the frontal cortex, caudate nucleus and putamen; and (2) some of the amphetamine-induced changes in CMRglu correlated with D amphetamine plasma levels. The present findings of cortical and subcortical increases in cerebral metabolism after D-amphetamine application in humans accord with previous studies in animals, demonstrating that relatively high doses of D amphetamine (presumably at least 1 mg/kg) are needed to increase cerebral glucose metabolism. PMID- 9849725 TI - Cerebral blood flow changes in patients with conversion disorder. AB - We evaluated regional cerebral blood flow in five patients with conversion disorder (three females, two males, mean age+/-S.D.: 29.8+/-9.5 years) with astasia-abasia. The patients underwent single photon emission computed tomography after the injection of 555 MBq of [99mTc]hexamethylpropyleneamine oxime. Uptake ratios between areas of decreased perfusion and normal brain regions were considered significantly decreased when there was a change > or = 10%. Four of the five patients had left temporal and one patient had left parietal perfusion decreases. Uptake ratios ranged from 0.72 to 0.88 (mean+/-S.D.: 0.81+/-0.08). Our findings suggest that alterations in regional brain perfusion may accompany conversion symptoms. Functional imaging may therefore offer a means of elucidating the neural correlates of conversion disorder. PMID- 9849726 TI - Changes in regional cerebral blood flow demonstrated by single photon emission computed tomography in depressive disorders: comparison of unipolar vs. bipolar subtypes. AB - Single photon emission tomography (SPECT) with 99mTc-HMPAO was used to compare regional cerebral blood flow (rCBF) in patients with unipolar and bipolar depression. The study group consisted of 10 unipolar depressed patients and seven bipolar depressed patients who met the DSM-III-R criteria for major depressive disorder (MDD). Nine physically and mentally healthy volunteers served as control subjects. SPECT images were obtained in the patients at two time points: (1) during the major depressive episode before patients had received medication; and (2) at the beginning of the remitted state while patients were receiving antidepressant medication. During the depressive episode, unmedicated unipolar depressed patients showed relatively increased left frontal rCBF compared both with the control subjects and the bipolar patients (P < 0.05). No significant differences in rCBF emerged between the bipolar patients and the control subjects. The data suggest that unipolar depressed patients, unlike bipolar patients, have relatively increased rCBF in the left frontal lobes during the depressive episode, but these differences tend to disappear during the period of remission. PMID- 9849727 TI - Clinical studies on three cases of the interval form of carbon monoxide poisoning: serial proton magnetic resonance spectroscopy as a prognostic predictor. AB - Three patients with the interval form of carbon monoxide (CO) poisoning were studied for associations between their clinical courses and serial changes on: (1) MRI; (2) EEG; (3) single photon emission computed tomography with N-isopropyl p-[123I]iodoamphetamine ([123I]IMP SPECT); and (4) proton magnetic resonance spectroscopy ([1HIMRS) to evaluate their usefulness as prognostic predictors. A hyperintense area on MRI T2-weighted images and a hypointense area on T1-weighted images, appearing in the deep white matter, persisted for a prolonged period even after improvement of the clinical symptoms, and did not become an accurate indicator of clinical evaluation or prognosis. [1H]MRS studies with the volume of interest set in the frontal lobe white matter revealed increases in choline containing compounds, and reductions of N-acetylaspartate in all cases. These findings normalized in one subclinical case over time. Distinctive findings in the severe cases included increased lactate in two cases showing akinetic mutism, with a difference in prognosis noted according to the degree and period of persistence of this increase. EEG findings were relatively well correlated with clinical symptoms in the early period, with a good correlation no longer consistent after a certain period was exceeded. [123I]IMP SPECT findings did not always reflectclinical symptomatology either. These results indicate that [1H]MRS is the most useful indicator in the clinical evaluation of patients with the interval form of CO poisoning. PMID- 9849728 TI - Surgical management of left colon obstruction: the University of Minnesota experience. AB - BACKGROUND: Management of left-sided colonic obstruction is a surgical challenge. This study was performed to review our management of patients with left colon obstruction presenting to the University of Minnesota Hospitals over a 10-year period, 1985 to 1994. STUDY DESIGN: We did a retrospective chart review of 143 patients (48 male and 95 female; mean age 70 years). RESULTS: Sites of obstruction were rectosigmoid, 40%; sigmoid colon, 47%; descending colon, 5%; and splenic flexure, 8%. Fifty-two percent of patients had obstructing colorectal cancer. Two patients presented with generalized peritonitis secondary to colonic perforation. The majority (n = 121, 85%) of patients underwent resection (subtotal in 39 [32%], and segmental in 82 [68%]) and anastomosis in a single stage after appropriate resuscitation. Intraoperative colonic cleansing was undertaken in 40 patients (28%). Morbidity within 30 days of operation was 11%, including 1 anastomotic leak, and mortality was 3%. The 4 deaths occurred in patients over 75 years of age and were not from anastomotic complications. CONCLUSIONS: A single stage resection and an anastomosis facilitated by intraoperative colonic cleansing in one-third of cases was performed in 85% of patients presenting with left colon obstruction. One anastomotic leak occurred. Our current policy of strongly favoring a single stage, definitive operation for patients presenting with left colon obstruction appears reasonable on the basis of this retrospective review of our experience. PMID- 9849729 TI - Longterm outcomes and quality of life after Z-shaped anastomosis for Hirschsprung's disease. AB - BACKGROUND: Z-shaped anastomosis is one of the modifications of Duhamel's procedure that was designed to eliminate the blind rectal pouch and to achieve complete resection of the colorectal septum. It has been the most widely performed operation in Japan for many years. The longterm postoperative function of evacuation and quality of life of the patients are considered important to evaluate this procedure. METHODS: At Kyushu University Hospital, from 1963 to 1997, 127 patients with Hirschsprung's disease underwent Z-shaped anastomosis. As a result, 122 out of 127 patients (96%) survived. The present status and symptoms, and anorectal functions, including a manometric study and barium enema, were evaluated during the clinical followup. RESULTS: A total of 99 of the 122 surviving patients (81%) were available for this study, and the mean postoperative period was 16 years. Evacuation scores in all patients were as follows; excellent, 62.2%; good, 28.6%; fair, 8.2%; and poor, 1.0%. The percentage of the patients who showed severe symptoms was 4.1% for diarrhea, 3.1% for constipation, 5.1% for incontinence, and 7.1% for soiling. The evacuation score improved chronologically and tended to reach a plateau at 10 to 15 years after operation, at which time 73% of the patients showed excellent outcomes and 95% were satisfactory (good or excellent). The appearance of a sense of defecation and an increase in the pressure difference between the anal canal and the rectum substantially contributed to the improvement in the defecation score. The appearance of the rectosphincteric reflex, including the atypical one, was seen in 40.5% of patients, but the appearance of a reflex did not seem to be related to the clinical status of defecation. Twenty-two of 30 patients older than 20 years were married, and 8 patients had children. CONCLUSIONS: The evacuation scores of in-patients undergoing Z-shaped anastomosis improved with age and were satisfactory (good or excellent) in most patients at least 10 years after operation. Most of the patients adapted to a normal social life. PMID- 9849730 TI - The prediction of common bile duct stones using a neural network. AB - BACKGROUND: The role of preoperative ERCP and endoscopic sphincterotomy (ES) in the diagnosis and treatment of suspected common bile duct stones (CBDS) in the laparoscopic age is controversial. The preoperative diagnosis of CBDS by ERCP and the removal of CBDS by ES are advantageous because of technical difficulties in performing laparoscopic exploration of the common bile duct. Approximately 50% of preoperative ERCP examinations are normal, however. The noninvasive diagnosis of CBDS has assumed new importance, but it has proved to be an elusive goal. Neural networks are a form of artificial computer intelligence that have been used successfully to interpret ECGs and to diagnose myocardial infarcts. The purpose of this study was to determine whether a neural network could be trained to predict CBDS accurately in patients at high risk of having duct stones. STUDY DESIGN: We trained a back-propagation neural network to predict the presence of CBDS. Retrospective data from patients who had a cholecystectomy and either a preoperative ERCP or intraoperative cholangiogram were used to build the network, and it was tested using unseen data. RESULTS: One hundred forty patients were used to train the network, and 16 patients were used to test it. The trained network was able to predict CBDS in 100% of the patients in both the training and test sets. CONCLUSIONS: Screening of high-risk patients for CBDS by neural network analysis is highly accurate. This promising new, noninvasive, and inexpensive technique can potentially decrease the need for preoperative ERCP by 50%, but additional prospective evaluation is indicated. PMID- 9849731 TI - Results of a technique of pancreaticojejunostomy that optimizes blood supply to the pancreas. AB - BACKGROUND: Anastomotic failure after pancreaticojejunostomy is still a common problem. Failure rates have not decreased perceptibly in the past 3 decades. The neck of the pancreas is a vascular watershed between celiac and superior mesenteric arterial systems. Prior attempts to reduce anastomotic failure at pancreaticojejunostomy have not focused on issues related to blood supply of the pancreas. The aim of this study was to determine whether pancreaticojejunostomy performed using a technique that included optimization of blood supply to the pancreas, would result in a low anastomotic failure rate. METHODS: The technique was prospectively evaluated in 40 patients having pancreaticojejunostomy, 39 during pancreaticoduodenectomy and 1 after traumatic transection of the neck of the pancreas. Blood supply to the pancreatic neck was evaluated clinically and by Doppler techniques. When blood supply was considered marginal, the pancreas was re-resected 1.5-2.0 cm to the left, away from the vascular watershed. RESULTS: Blood supply at the cut margin of pancreas was judged as brisk in 24 patients and marginal in 16 patients. Resecting a segment of pancreas in these 16 patients resulted in brisk bleeding from the new cut margin in all but 1 patient who had an anomalous artery that had to be sacrificed for oncologic reasons. The only fistula in the series occurred in this patient. There were no intraabdominal abscesses. CONCLUSIONS: A technique that includes ensuring adequate blood supply to the pancreas can result in a very low rate of anastomotic failure. PMID- 9849732 TI - The number of metastatic lymph nodes: a promising prognostic determinant for gastric carcinoma in the latest edition of the TNM classification. AB - BACKGROUND: The number of metastatic regional lymph nodes determines the new pN categories in the 5th edition of the TNM classification. STUDY DESIGN: Our retrospective study was conducted to compare the new method of defining lymph node status with the conventional classification, consisting of the anatomic extent of lymph node metastases, a well-established prognostic factor. The study was based on clinical data for 493 patients with gastric carcinomas who underwent potentially curative operations and had histologically confirmed nodal metastases. These patients were stratified into 1) n categories according to the Japanese Classification of Gastric Carcinoma, 2) the new pN categories, and 3) the pN categories determined by the number of metastatic perigastric nodes resected by standard D1 gastrectomy. Survival data were analyzed for each group. RESULTS: The number of metastatic nodes after D2 lymphadenectomy reflected prognosis well and was shown by multivariate analysis to be a strong independent prognostic factor. When the classification was performed limited to the metastatic perigastric nodes, stage migration was evident, but the variable remained competent as a prognostic indicator. CONCLUSIONS: The number of metastatic nodes is a promising determinant in the new international stage classification. PMID- 9849733 TI - Image-guided or needle-localized open biopsy of mammographic malignant-appearing microcalcifications? AB - BACKGROUND: The evaluation and initial management of abnormalities detected on screening mammography have evolved substantially over the last decade. This study was designed to evaluate the most appropriate initial diagnostic biopsy technique for patients presenting with malignant-appearing microcalcifications on screening or diagnostic mammography. STUDY DESIGN: An institutional review of a prospective database was performed to compare initial image-guided breast biopsy (IGBB) and needle-localized open biopsy (NLOB) in patients presenting with malignant appearing microcalcifications. Patients with atypical hyperplasia (AH) or carcinoma in situ (CIS) were identified and reviewed separately. Measures of outcomes included the total number of procedures, time from initial biopsy to definitive treatment, charges, and percentages of patients who required both procedures. RESULTS: A total of 17,121 patients underwent mammography from July 1994 to December 1996 at Gundersen Lutheran Medical Center. Indeterminate microcalcifications were found in 167 patients and were the reason for IGBB in 112 and NLOB in 55 patients. Histologic results included 81 patients (48%) with benign lesions, 25 (15%) with invasive cancers, and 61 (37%) having a proliferative finding including AH or CIS. Ductal CIS was present in 42 (72%) of the 61 proliferative lesions. Comparisons were made between the groups of patients with CIS or AH who underwent initial NLOB (n = 25) versus those having initial IGBB that was followed by a secondary NLOB (n = 25). The median elapsed time to definitive therapy was 20 days (range 0 to 336 days) for initial IGBB followed by NLOB and 7 days (range 0 to 79 days) for an initial NLOB performed for suspicious microcalcifications (p = 0.0367). The total number of procedures performed on each patient and total costs were also less for patients having an initial NLOB. CONCLUSIONS: The time to definitive local therapy, the number of procedures, and overall charges were less for patients with AH or CIS having initial NLOB as opposed to initial IGBB. Careful initial evaluation of microcalcifications may identify some patients for whom an initial NLOB remains the most appropriate procedure. Such patients desiring breast-conserving therapy may benefit in terms of time to definitive treatment, total number of procedures performed, and cost if a careful NLOB is the initial procedure performed as a formal lumpectomy. PMID- 9849734 TI - Cosmetic results after wire-guided biopsy of benign breast lesions. AB - BACKGROUND: There are surprisingly few reports about the cosmetic results of wire guided biopsy of benign breast lesions as opposed to breast-conserving surgery and irradiation of early breast cancer (BCT). STUDY DESIGN: Twenty potential perioperative risk factors for adverse cosmetic results after wire-guided breast biopsy were prospectively evaluated in 101 patients undergoing the first single biopsy after suspicion of a malignant lesion that subsequently proved to be benign. The overall cosmetic result was evaluated by using 6 specific cosmetic indices individually scored 6 months after the breast biopsy. RESULTS: The overall cosmesis was excellent, good, fair, or poor in 48.5%, 26.7%, 12.9%, and 11.9% of cases, respectively. The corresponding figures according to appraisal by the patients were 22.8%, 58.4%, 17.8%, and 1.0%, respectively. Unsatisfactory (fair or poor) overall cosmetic results were related to excisions extending down to the fascia (p = 0.001) and postoperative complications (p = 0.018) in multivariate analysis. Notably, specimen volume had no significant impact on overall cosmesis, as opposed to cosmesis after BCT. CONCLUSIONS: Cosmetic outcomes after wire-guided biopsy of benign breast lesions were excellent or good in at least 75% of cases. Excisions extending down to the pectoralis fascia and complications were associated with poor aesthetic outcomes. PMID- 9849735 TI - Erythropoietin therapy improves graft patency with no increased incidence of thrombosis or thrombophlebitis. AB - BACKGROUND: Recombinant human erythropoietin (rHuEPO) for the treatment of severe anemia in patients with end-stage renal disease (ESRD) is suggested to improve rehabilitation and cognitive function. The criticism is the alleged increase in the failure rate of arteriovenous (AV) access grafts and in the incidence of lower-extremity deep venous thrombophlebitis (DVT). This study addressed the longevity of AV grafts and the incidence of DVT. STUDY DESIGN: We reviewed 481 consecutive patients with ESRD on dialysis with PTFE access grafts, including 173 consecutive patients who were receiving rHuEPO and 308 who were not. rHuEPO was administered during dialysis titrated against the hematocrit to achieve a level of 33% to 38%. The rHuEPO-ESRD group included 173 patients with a mean age of 58 years, including 54% women; 84% of the grafts were in the upper extremity. In the control group of 308 patients, 57% were women. Diabetes and hypertension were controlled in both groups. RESULTS: Forty-five of 173 rHuEPO patients (26%) experienced graft thrombosis within 1 year. Among 88 episodes of thrombosis, 14 patients experienced multiple episodes. Primary patency was 8.9 months; secondary patency was 11.2 months. In the control population, 95 of 308 patients (31%) experienced graft thrombosis; 27 patients had multiple episodes. Primary patency was 7.8 months and secondary patencywas 9.8 months. The hematocrit improved from a mean of 23% in the control group to 34% in the treated rHuEPO group. Two patients in the control group and one patient receiving rHuEPO experienced DVT in the lower extremity. CONCLUSIONS: Primary and secondary AV fistula patency rates were improved by 10% with rHuEPO. rHuEPO did not increase DVT. PMID- 9849737 TI - Occult injuries to the diaphragm: prospective evaluation of laparoscopy in penetrating injuries to the left lower chest. AB - BACKGROUND: To evaluate the incidence of occult diaphragmatic injuries and investigate the role of laparoscopy in patients with penetrating trauma to the left lower chest who lack indications for exploratory celiotomy other than the potential for a diaphragm injury. STUDY DESIGN: Patients with penetrating injuries to the left lower chest who were hemodynamically stable and without indications for a celiotomy were prospectively evaluated with diagnostic laparoscopy to determine the presence of an injury to the left hemidiaphragm. Diagnostic laparoscopy was performed in the operating room under general anesthesia. RESULTS: One-hundred-ten patients (94 stab wounds, 16 gunshot wounds) were evaluated with laparoscopy. Twenty-six (24%) diaphragmatic injuries were identified (26% for stab wounds and 13% for gunshot wounds). Comparison of patients with diaphragmatic injuries with those without diaphragmatic injuries demonstrated a slightly greater incidence of hemo/pneumothoraces (35% versus 24%, NS). The incidence of diaphragmatic injuries in patients with a normal chest x ray was 21% versus 31% for patients with a hemo/pneumothorax. An elevated left hemidiaphragm was associated with a diaphragmatic injuries in only 1 of 7 patients (14%). The incidence of diaphragmatic injuries was similar for anterior, lateral, and posterior injuries (22%, 27%, and 22% respectively). CONCLUSIONS: The incidence of occult diaphragmatic injuries in penetrating trauma to the left lower chest is high, 24%. These injuries are associated with a lack of clinical and radiographic findings, and would have been missed had laparoscopy not been performed. Patients with penetrating trauma to the left lower chest who do not have any other indication for a celiotomy should undergo videoscopic evaluation of the left hemidiaphragm to exclude an occult injury. PMID- 9849736 TI - Perioperative outcomes of major hepatic resections under low central venous pressure anesthesia: blood loss, blood transfusion, and the risk of postoperative renal dysfunction. AB - BACKGROUND: We have previously demonstrated that maintenance of a low central venous pressure (LCVP) combined with extrahepatic control of venous outflow reduced the overall blood loss during major hepatic resections. This study examined the overall outcomes and, in particular, renal morbidity associated with a large series of consecutive major liver resections performed with this approach. In addition, the rationale for the anesthetic management to maintain LCVP was carefully reviewed. STUDY DESIGN: All major hepatectomies performed between December 1991 and April 1997 were reviewed. The prospective Hepatobiliary Surgical Service database was merged with the Memorial Hospital Laboratory and Blood Bank databases to yield the nature of the operation, blood loss, blood product transfusions, outcomes, and levels of preoperative, postoperative, and discharge serum creatinine and blood urea nitrogen. RESULTS: A total of 496 LCVP assisted major liver resections were performed, with no intraoperative deaths and an in-hospital mortality rate of 3.8%. The median blood loss was 645 mL. Sixty seven percent of the patients did not require perioperative blood transfusion during surgery and the immediate 12 hours after surgery. The median number of blood transfusions was 2. Only 3% of the patients experienced a persistent and clinically significant increase in serum creatinine possibly attributable to the anesthetic technique. Renal failure directly attributable to the anesthetic technique did not occur. CONCLUSIONS: Major resection with LCVP allowed easy control of the hepatic veins before and during parenchymal transection. The anesthetic technique, designed to maintain LCVP during the critical stages of hepatic resection, not only helped to minimize blood loss and mortality but also preserved renal function. PMID- 9849738 TI - Restorative colectomy for large bowel obstruction: a new paradigm? PMID- 9849739 TI - Modified V-Y advancement technique for mastectomy closure. PMID- 9849740 TI - A technique for distal splenoadrenal shunting in pediatric portal hypertension. AB - Technical modification of the DSRS by using the end-to-end splenoadrenal anastomosis allows for effective selective decompression of portal hypertension in children, can be accomplished with no perioperative mortality, and has demonstrated longterm patency and minimal morbidity. Clinical encephalopathy was seen in only 1 patient although 3 additional patients required lactulose therapy. PMID- 9849741 TI - Ventriculoperitoneal shunt placement with video-laparoscopic guidance. PMID- 9849742 TI - Low molecular weight heparins: current use and indications. AB - Thromboembolic complications are associated with significant morbidity and mortality in postoperative patients. For many years, unfractionated heparin has been used successfully in primary and secondary prophylaxis of these complications. In recent times, however, the usefulness of LMWHs has caught the attention of clinicians because of improved bioavailability, predictable anticoagulation, ease of administration, and the lack of need for monitoring anticoagulation. In clinical situations, LMWHs have been tested and proved to be safe and equipotent or supe rior when compared with unfractionated heparin or warfarin (Table 5). It is clear from clinical trials that LMWHs are superior in primary prophylaxis of DVT in orthopaedic surgical procedures, treatment of unstable angina, and in patients with multiple traumas. LMWHs were also tested and found to be an acceptable alternative to unfractionated heparin in both the primary prophylaxis of DVT in high risk general surgical procedures and in the treatment of patients with DVT and pulmonary embolism. However, the role of LMWHs in ischemic heart diseases, valvular heart diseases, postcoronary angioplasty, and vascular surgery remains to be proved. The major impact of LMWHs would be in allowing clinicians to treat PE and DVT in an outpatient setting, which would directly impact medical economics. LMWHs are associated with similar complications as unfractionated heparin is, but the complications occur less frequently. Currently, the main limitation in using LMWHs in place of unfractionated heparin or warfarin is its cost. However, taking into account the cost incurred by hospitalization and longterm monitoring of anticoagulation in patients treated with unfractionated heparin, certain trials have proved the cost of LMWHs to be the same or less than the cost of unfractionated heparin overall. We envision that LMWHs will be widely used in the future and will bring welcomed change in the treatment of thromboembolic diseases. PMID- 9849743 TI - Proceedings of the 7th Annual Meeting of the Association for Applied Human Pharmacology (AGAH) e.V, Bad Honnef, 25-27 January 1998. PMID- 9849744 TI - Clinical pharmacology in the target population, Annual Meeting of AGAH, 1998. Human pharmacology, clinical pharmacology: common aims and position of the Society for Applied Human Pharmacology (AGAH eV). PMID- 9849745 TI - Human pharmacology -- clinical pharmacology. Common goals and tasks from the point of view of the DGPT Section Clinical Pharmacology (German society of experimental and clinical Pharmacology and Toxicology). PMID- 9849746 TI - Effect of age on pharmacokinetics and pharmacodynamics in man. AB - Elderly patients represent an increasing part of our population who consume disproportionately high amounts of drugs. During aging physiological and disease induced changes occur which might affect PK and/or PD of many drugs. The calcium channel blocker verapamil and the benzodiazepine midazolam were taken as examples to illustrate some problems and open questions in research pertaining to the elderly. Following an acute i.v. dose of racemic verapamil and during steady state (120 mg bid p.o.) no significant differences of the stereoselective disposition of S- and R-verapamil could be found between younger and older healthy subjects. Concomitant intake of rifampicin (600 mg/die) induced especially the presystemic (prehepatic) metabolism of verapamil so that oral bioavailability approached zero and PD effects almost diminished. Intestinal metabolism and inducibility were well preserved in the elderly. The PK and PD of midazolam were investigated in young and older patients who received an i.v. bolus of 0.05 and 0.03 mg/kg, respectively, for premedication prior to third molar teeth extraction. Again, no significant differences in the PK parameters were found between both groups. However, the sedative effects were much more pronounced in the elderly and this population demonstrated a significantly higher CNS-sensitivity to midazolam. All physicians should be aware that for various reasons drug response can be age-dependent and therefore more PK-PD data are needed for the elderly. PMID- 9849747 TI - Influence of gender on the pharmacokinetics and pharmacodynamics of drugs. AB - Historically, women, the elderly, and minorities were underrepresented in clinical drug trials. Information on possible gender-related differences in the pharmacokinetics of drugs is often lacking, although for some drugs significant differences could be demonstrated. In women, absorption, protein binding, volume of distribution, and metabolism of drugs may differ due to hormonal influences on physiological functions. Sex-related differences could be shown for phase I (cytochrome P450) as well as phase II (especially glucuronidation) reactions. Since many women world-wide take oral contraceptives, data should be provided to determine to what extent other drugs are influenced by estrogens and progestogens or to what extent the other drugs may attenuate the contraceptive efficacy. Moreover, estrogens interact with various enzymes and receptors, e.g. at the endothelial function as well as dopaminergic receptor sites, and may therefore attenuate or enhance drug effects or even drug side-effects. For a number of drugs it is well recognized, that women suffer more frequently from side-effects, however, it is often not clear, if this is due to gender differences in the pharmacokinetics or pharmacodynamics of the responsible drug. Very little is known about these gender-related differences and the possibility that women may show a different pattern of treatment response than men. As a result, drug approval authorities now require more data on the pharmacokinetics of novel drugs in women as well as a sufficient accrual of women in efficacy and outcome trials. PMID- 9849748 TI - Discussion on early clinical drug development in the target population with respect to the field of reproductive endocrinology. AB - The study populations taking part in early clinical drug development of reproductive endocrine treatments are discussed. After having compared subjects in phase I studies with subsequent target populations, the question was posed as to whether the inclusion of the later target population could accelerate the developmental process. This applies only to certain specific clinical questions that are posed in phase I studies. From the authors' perspective, the key goal of rapid and well-structured early drug development is to be attained by using reliable surrogate markers that are able to reflect the clinical effects. PMID- 9849749 TI - Pharmacokinetic studies in volunteers with renal impairment. AB - According to Luzius Dettli, drug clearance is a linear function of renal function. The slope of this function can be predicted from the fraction of a drug that is eliminated by the renal route. Pharmacokinetics in patients with functional anuria can, however, considerably deviate from these predictions. All basic pharmacokinetic parameters depend differently and specifically on renal function: drug clearance and distribution volume depend on creatinine clearance, but elimination half-life and plasma-binding correlate better with serum creatinine. For drugs with saturable tubular secretion, it can be shown that drug clearance depends on creatinine clearance in accordance with a left-bent, convex function, but not in accordance with a right-bent, concave function. It might be reasonable to postulate that the pharmacokinetics of every new approved drug should be determined in individuals with renal impairment. For drugs with no severe risks of adverse effects, these studies can be performed in volunteers with renal impairment better than in patients. PMID- 9849750 TI - Phase I studies in patients with liver cirrhosis. PMID- 9849751 TI - Tobacco smoking, alcohol, and special diet in drug trials: exclusion criteria or target of specific investigations? PMID- 9849752 TI - Relevance of pheno- and genotyping in clinical drug development. AB - Individual variability in the plasma concentration of a xenobiotic is a considerable problem in the clinical use as well as in the clinical development of a new drug. In clinics altered drug response and drug toxicity are predominant findings. In clinical development (especially in the early phases) problems with the interpretation of individual pharmacokinetics and appearance of unexpected drug reactions may occur. One major reason for the inter-and intraindividual variability is based on variations in the metabolism of the drug that are dependent on the genetic variations of the metabolising enzymes. However, while the first of these so-called polymorphisms has already been described in 1979 (for a review see [Daly 1995]) and knowledge of these polymorphisms are still growing, the impact on clinical practice as well on the selection and development of drug candidates in the pharmaceutical industry at present is still limited. This may change in the near future as recent advances in molecular biology and especially in the diagnosis of individual genomic characteristics will result in a better understanding of the basic principles of the polymorphisms. High throughput screening methods will reveal information on the distribution of these polymorphic alleles in the target population and enable the broad characterization of the drug candidate in in vitro systems. The early knowledge as to whether a polymorphic pathway is involved in drug metabolism/action and of its clinical relevance will lead to a reduction of time and costs in the development of a new drug. PMID- 9849753 TI - Drug trials in pediatric oncology: a multi-facetted problem. AB - In the field of childhood malignancies, the last 30 years have witnessed the advance of highly effective treatment regimens and high cure rates through the clinical application of a growing number of cytotoxic agents. The therapeutic success was mainly brought about by the empirically based joint efforts of cooperative clinical trials that employed effective cytotoxic drug combinations at the threshold of maximum tolerable toxicity. Systematic clinical drug trials, however, aimed at the approval of specific substances for use in pediatric oncology have been lacking. The following presentation will try to describe briefly the various facets of the problem involved in the systematic development of drugs for childhood malignancies as is manifested in Germany today. PMID- 9849754 TI - Clinical trials on biologics in rheumatoid arthritis. AB - BACKGROUND: Cytokines and T cells play a major role in the pathogenesis of rheumatoid arthritis (RA). Biologic targeting is a novel therapeutic approach. Published trials in humans are discussed in this paper. METHODS: CD4-positive T cells, proinflammatory cytokines like TNF-alpha, IL-1, IL-6 and gamma-IFN were major targets for therapy. Biologics were constructed of monoclonal human and non human antibodies, chimerics of both, antiinflammatory regulatory proteins like IL 1 receptor antagonist, IL-10, and fusion proteins consisting of receptors and immunoglobulins. RESULTS: More than 2000 humans with RA were exposed to biologics in the last decade. Both, toxic and safe, efficient and non-efficient drugs were tested. Phase II data could not confirm preliminary phase I data in several drugs tested. The pharmacokinetic profile of biologics is influenced by frequent induction of human anti drug antibodies. CONCLUSION: The major role of cytokines in RA has been confirmed. Due to the limited long-term experience immunomodulation does not replace conventional pharmacotherapy. PMID- 9849755 TI - Primary slings for everyone with genuine stress incontinence? The argument for... PMID- 9849756 TI - Vesicouterine fistula: an updated review. AB - To ascertain the condition of vesicouterine fistula, a search of the medical literature using the Ovid version (Ovid Technologies Inc., New York, USA) of Medline (1966-1997) was undertaken, using the keywords vesicouterine, vesico uterine, uterovesico, and utero-vesico fistula. A search of the earlier medical literature was by cross-referencing and is likely to be less complete than the computerized database. Nonetheless, the authors believe the most relevant publications were cited. On reviewing the subject it was noticed that there had been a change in obstetric practice between the first and the latter half of this century, namely an increase in the incidence of cesarean deliveries. However, the literature contains relatively little information on pathophysiology, possibly owing to the relative rarity of the lesion. Emphasis has been given to the most recent 10 years (1987-1997) in order to update the etiology, diagnosis and management of this clinical condition. PMID- 9849757 TI - A randomized controlled trial of pelvic floor muscle exercises to treat postnatal urinary incontinence. AB - A randomized controlled trial was carried out to evaluate the extent to which a program of reinforced pelvic floor muscle exercises (PFME) reduces urinary incontinence 1 year after delivery. Two hundred and thirty women who were incontinent 3 months postpartum were randomized to either a control group doing standard postnatal pelvic floor muscle exercises (n = 117) or to an intervention group (n = 113) who saw a physiotherapist for instruction at approximately 3, 4, 6 and 9 months postpartum. Results collected 12 months after delivery included prevalence and frequency of incontinence and PFME, sexual satisfaction, perineometry measurements and pad tests. Twenty-six (22%) of the control group and 59 (52%) of the intervention group withdrew before the final assessment. The prevalence of incontinence was significantly less in the intervention group than in the control group (50% versus 76%, P=0.0003), and this group also did significantly more PFME. There were no significant differences between the groups as regards sexual satisfaction, perineometry measurements or pad test results. PMID- 9849758 TI - Pelvic floor electrical stimulation for genuine stress incontinence: who will benefit and when? AB - This study sought to determine the characteristics of women in whom pelvic floor electrical stimulation will reduce stress urinary incontinence. It also evaluates how long electrical stimulation should be used before significant improvements are seen in clinical outcomes. Subjects with genuine stress incontinence were enrolled into a multicenter non-randomized trial. They used electrical stimulation for 15 minutes twice daily or every other day for 20 weeks. At the end of 20 weeks, those with a 50% reduction in leakage episodes on voiding diary ('responders') were compared with those who did not show a 50% reduction ('non responders'). Thirty-one subjects were enrolled and 28 completed the study. After the treatment period, 19 subjects were defined as responders and 9 as non responders. There were no significant differences between the two groups in baseline demographics (e.g. age, parity, largest birth weight etc.) other than body mass index (greater in nonresponders). Significant subjective and objective improvements were noted among responders by 10 and 14 weeks, respectively. Compliance was higher in responders during weeks 12-15 of the study (P=0.05). It was concluded that a minimum of 14 weeks of pelvic floor stimulation was necessary before significant objective improvements were seen. Body mass index and patient compliance may affect success. PMID- 9849759 TI - Efficacy of an external urethral device in women with genuine stress urinary incontinence. AB - The efficacy of a new external anti-incontinence device in patients with a videourodynamic diagnosis of genuine stress incontinence (GSI) in an open longitudinal study is reported. Fourteen women with GSI underwent assessment before and after 3-4 weeks of device use. Assessment consisted of visual analog scores (VAS), quality of life (QOL) questionnaires, urine for culture and a 1 hour pad test. VAS scores showed a significant improvement for the symptom of stress incontinence (P<0.05). QOL scores improved significantly by 38% (P<0.05) and 29% (P<0.01) for the Incontinence Impact Questionnaire and Urogenital Distress Inventory, respectively. The mean pad weight decreased by 47% (P=0.056). Of the 9 women who had a positive pad test (>2 g) without the device, 5 were dry (<2 g) with the device (P<0.05). These preliminary data suggest that this device is effective in women with GSI. PMID- 9849760 TI - The use of small intestine in bladder reconstruction. AB - Reconstruction of the bladder is a treatment available to patients who have a diseased or damaged bladder, and small bowel is the most commonly used tissue. Augmentation cystoplasty increases the total bladder capacity, whereas substitution cystoplasty replaces the whole organ. This is either drained through a continent cutaneous stoma or is reanastomosed to the urethra as an orthotopic reconstruction. Although the treatment for invasive bladder cancer has not changed greatly in the last few decades, the use of orthotopic bladder reconstruction allows for a great improvement in the quality of life for patients who undergo cystectomy. These reconstructive techniques can also be offered to patients with other forms of pelvic malignancy that involve the bladder. PMID- 9849761 TI - Electrical stimulation for stress incontinence. AB - Electrical stimulation has been reported to be effective for stress incontinence, cure and improvement rates being reported to range from 30% to 50%, and from 6% to 90%, respectively. However, clinical application of this treatment is not common because there is little physiological and technical information. Electrodes for electrical stimulation are divided into two types: external (non implantable) and internal (implantable), and there are two methods of stimulation: chronic (long-term, continuous) and short-term. Frequencies of 20-50 Hz, with a pulse duration of 1-5 ms, have been reported to be effective for urethral closure. The effectiveness of the treatment should be verified with placebo-controlled double-blinded trials, and four such studies using an active and a sham device have been reported. Two of these verified the superiority of the active device over the sham device, but the others did not demonstrate any significant difference between the two with regard to efficacy. Electrical stimulation has been reported to result in a long-term continuation of therapeutic effect. The effect has been explained as a re-education or a reactivation of lost functions of the pelvic floor muscles. As to adverse effects, there may be some complications in relation to anesthesia or surgical procedures, such as infection, pain and bleeding with implantable electrodes. The incidence of adverse effects in short-term electrical stimulation is less than 14%. In conclusion, short-term electrical stimulation using non-implantable anal or vaginal electrodes is the most recommendable because of safety and ease of use. PMID- 9849762 TI - Electrophysiological properties of the bladder. AB - The electrophysiological properties of detrusor smooth muscle are described, in particular with regard to their influence on the contractile properties of the tissue. The Ca2+ and K+ channel activities are most important in generating action potentials, but the role of several other ionic currents is described, including Cl-, Ca2+-activated, stretch-activated and ligand-gated channels. The variable appearance and functions of different ionic currents in disease states is discussed, as well as the question of whether electrical activity can transmit between adjacent smooth muscle cells. In addition, the precise role that electrophysiological phenomena play in the regulation of the contractile state of the smooth muscle cells, as well as the generation of bladder electromyograms, is discussed. PMID- 9849763 TI - Application of the concept of 'therapeutic index' to surgery for stress incontinence. AB - The pharmacologic concept of therapeutic index can be applied to surgery for stress incontinence to obtain an objective prediction of surgical outcome. The rationale and mechanics of its application to surgery are discussed in this article. The conclusions support current clinical trends which favor discarding the anterior repair and needle suspension procedures as first-line surgical treatment of stress urinary incontinence in favor of the retropubic procedures. Suburethral sling procedures should be, as now, reserved for second-line surgical treatment of stress urinary incontinence. PMID- 9849764 TI - An unusual foreign body in the bladder 7 years after a Stamey endoscopic bladder neck suspension. AB - Delayed migration of the suture and bolster after an endoscopic bladder neck suspension across tissue planes, with subsequent erosion into the bladder, is uncommon. We present a case of late migration of the suture and bolster occurring 7 years after a Stamey endoscopic bladder neck suspension. A 56-year-old woman had undergone a Stamey procedure in June 1990. In January 1997 she presented with discomfort in the left iliac fossa and the groin. A midstream sample of urine showed microscopic hematuria. Imaging and endoscopic examinations revealed a calcified lesion on the left lateral wall of the bladder, attached to the Stamey sutures. Cystolitholapaxy was attempted, but during the procedure it became obvious that there was a calcified cuff attached to the suture. This was removed endoscopically, along with its suture. Cystoscopy should be considered early in the evaluation of patients presenting with lower abdominal discomfort or irritative voiding symptoms after retropubic bladder neck suspension. PMID- 9849766 TI - Readmissions to drug abuse treatment and HIV risk behavior. AB - The objectives of the study were (a) to investigate the characteristics of drug abuse treatment clients who return to treatment and (b) among those with readmissions, to describe changes over time in risk behavior for human immunodeficiency virus (HIV) infection and to identify factors associated with behavior change. Data were derived from a multisite HIV surveillance program in a single community; the program used a unique identifier to link HIV test results and behavioral information from multiple contacts. During a 30-month period, 1994 clients were admitted to three satellite facilities of a single treatment agency: detoxification, long-term residential, and outpatient. Of these clients, 574 (29%) had one or more readmissions to the same or a different facility during the 24 months following the index admission. Drug injectors, those tested for HIV, and those living in the community were more likely to be readmitted to treatment. There was little overall change in HIV risk behavior between the index admission and the readmission furthest in time from the index admission. Clients whose index visit was at the residential facility were more likely to reduce their injection risk behavior than those admitted to the other facilities. Clients readmitted to either the residential or the outpatient facility were more likely to have reduced their injection risk behavior than those readmitted to detoxification. Treatment facility was not associated with sexual risk behavior change. Men were more likely than women to reduce their high-risk sexual behaviors. The results underscore the need for treatment programs to make HIV testing readily available to their clients and to make special efforts to assist female clients to reduce their HIV risk. PMID- 9849767 TI - Substance use and abuse among patients with comorbid dysthymia and substance disorder. AB - This study determines the substance use and abuse patterns among patients with comorbid substance-related disorder (SRD) and dysthymia in SRD-dysthymia as compared with patients with SRD only. Differences in use and abuse patterns could be useful for (a) understanding motivations for use, such as self-treatment, and (b) assisting clinicians to identify cases of dysthymia among SRD patients. Retrospective and current data were obtained regarding history of substance use and current SRD diagnoses. Two university medical centers with alcohol-drug programs located within departments of psychiatry were the settings. A total of 642 patients was assessed. of whom 39 had SRD-dysthymia and 308 had SRD only. Data on past usc were collected by a research associate using a questionnaire. Current SRD and dysthymia diagnoses were made by psychiatrists specializing in addiction. The patients with SRD-dysthymia and SRD only did not differ with regard to use of alcohol, tobacco, and benzodiazepines. The patients with SRD dysthymia started caffeine use at an earlier age, had shorter "use careers" of cocaine, amphetamines, and opiates, and had fewer days of cocaine and cannabis use in the last year. They also had a lower rate of cannabis abuse/dependence. This study indicated that patients with dysthymia and SRD have exposure to most substances of abuse that is comparable to patients with SRD only. However, they selectively use certain substances less often than patients with SRD only. Early use of caffeine may reflect self-treatment for depressive symptoms among patients with SRD-dysthymia. PMID- 9849768 TI - The motivation for drug abuse treatment: testing cognitive and 12-step theories. AB - The purpose of this paper is to evaluate two models of behavior change: cognitive theory and 12-step theory. Research subjects were drawn from three separate, but parallel, samples of adults. The first sample consisted of out-of-treatment chronic drug users, the second consisted of drug users who had applied for treatment at a publicly funded multiple-provider drug treatment facility, and the third consisted of drug users who had applied for treatment at an intensive outpatient program for crack cocaine users. Cognitive theory was supported. Study participants applying for drug abuse treatment reported a higher level of perceived problem severity and a higher level of cognitive functioning than out of-treatment drug users. Two hypotheses drawn from 12-step theory were not supported. Treatment applicants had more positive emotional functioning than out of-treatment drug users, and one treatment-seeking sample had higher self-esteem. PMID- 9849769 TI - Prevalence of DSM-IV substance abuse and dependence disorders among prison inmates. AB - The study examined the 30-day and lifetime prevalence of DSM-IV alcohol and drug disorders among state prison inmates. A sample of 400 inmates consecutively admitted to a state prison reception center were assessed for alcohol and drug disorders using the Structured Clinical Interview for DSM-IV (SCID-IV). Test retest reliabilities were calculated for the SCID-IV. Lifetime substance abuse or dependence disorders were detected among 74% of inmates, including over half who were dependent on alcohol or drugs. For the 30 days prior to incarceration, over half of the sample were diagnosed as having substance abuse or dependence disorders, including 46% who were dependent on alcohol or drugs. Black inmates were significantly less likely to be diagnosed as alcohol dependent than whites or Hispanics. The high rates of substance use disorders are consistent with previous findings from other studies conducted in correctional settings and reflect the need to expand treatment capacity in prisons. PMID- 9849770 TI - Substance abuse and psychiatric illness: treatment experience. AB - The purpose of this paper is to describe an outpatient dual-diagnosis treatment program and 1-year clinical outcome and hospital utilization data. Subjects were 118 consecutive admissions to the Seattle Veterans Affairs (VA) Medical Center's Dual Disorders program over the period from June 1, 1992, to August 31, 1994. Program eligibility requirements included having a current substance use disorder and an active non-substance-related major Axis I disorder (typically major depression, post-traumatic stress disorder [PTSD], bipolar disorder, or schizoaffective disorder). The treatment frame involved group-based programming (including support, medications management, and psychoeducation), routine urine drug screening, and crisis interventions. Results showed that subjects averaged 1.5 non-substance-related Axis I psychiatric disorders (54% involving psychotic symptoms) and 1.8 active substance use disorders. Patients stayed engaged in treatment for a median of 217 days, with 60% of patients having no positive drug screens, and the overall sample having a 40% reduction in the number of inpatient bed days in the year after intake. Conclusions were that, for a number of patients with comorbid disorders, psychiatric stabilization and cessation of substance use can be accomplished within an outpatient treatment frame that averages two completed clinical contacts per week. PMID- 9849771 TI - Predicting postprimary treatment services and drug use outcome: a multivariate analysis. AB - The substance abuse treatment field is continually challenged to develop interventions that encourage drug abusers to remain longer in therapeutic services and that facilitate improved outcomes. As one of those interventions, case management has generally not been subjected to multivariate analysis to evaluate its role in accomplishing those goals. Using a sample of 444 veterans who received substance abuse treatment services, this study examines relationships among demographic and psychosocial variables at intake, assignment to either traditional or enhanced (case management) treatment services, and both proximal (postprimary treatment contact) and distal (severity of drug use) measures of outcome. Multivariate analyses reveal that case-managed clients stay longer in postprimary treatment services than non-case-managed clients, and consequently, longer postprimary treatment contact leads to better drug use outcomes. The implications of these findings are discussed. PMID- 9849773 TI - Hematological considerations in out-of-treatment drug users. AB - Complete blood counts (CBCs) were performed on 215 out-of-treatment injecting and noninjecting drug users participating in a federally sponsored human immunodeficiency virus (HIV) risk-reduction program in Houston, Texas. A substantial proportion of patients/clients were found to be suffering from varying degrees of anemia and other hematological conditions. Anemia is known to affect cognitive skills such as the ability to concentrate and process information. This impairment can limit the effectiveness of drug intervention and treatment, even among those users eager to participate in intervention or treatment programs. Consideration of substance users' physical condition as manifested by hematological measures can provide an important compliment to health behavioral models that stress cognitive functioning without consideration of the underlying physical processes involved. PMID- 9849772 TI - Relationship of cessation of cocaine use to cigarette smoking in cocaine dependent outpatients. AB - Cocaine abusers (N=43) from an outpatient drug rehabilitation program were interviewed regarding the relationship of cessation of cocaine use to cigarette smoking. Patients reported significant reductions in cigarette consumption after cessation of cocaine use (37.3+/-26.4 cigarettes per day when using cocaine vs. 18.1+/-7.0 cigarettes per day after cessation of cocaine use; P < .0001, paired t test, t=4.79 with 42 degrees of freedom). Reduction of cigarette smoking after cessation of cocaine use was reported primarily by heavy smokers. However, patients with longer clinic enrollments reported more increases and fewer reductions in the number of cigarettes smoked per day after cessation of cocaine use. Comparison of average craving scores from the most recent period of cocaine use (1 month to several years before the interview), at clinic enrollment (usually shortly after the most recent period of cocaine use), and during the 24 hours before the interview showed that patients experienced markedly decreased craving for cocaine, in contrast to only slightly decreased craving for cigarettes. PMID- 9849774 TI - Antisocial behavior, substance use, and somatization in families of adolescent drug abusers and adolescent controls. AB - Symptoms of conduct disorder (CD), antisocial personality disorder (ASP), alcohol abuse, drug abuse, and somatization were tabulated for the families of 35 delinquent, substance-abusing (multiple-problem) adolescent male probands and 35 age-matched control males. Alcohol abuse, CD/ASP, and somatization were assessed with either the Diagnostic Interview Schedule (DIS) (for participants aged 18 and up) or the Diagnostic Interview for Children and Adolescents (DICA) (for participants aged 12 to 17). Drug abuse for all participants was assessed with the Substance Abuse Module (SAM) of the Comprehensive Interview for Diagnostic Evaluation (CIDE). As expected, proband groups had significantly more (p < .0001) CD/ASP, alcohol abuse, and drug abuse symptoms than control groups. A significant (p < .01) positive correlation among CD/ASP, alcohol abuse, and drug abuse was found for each group. Somatization was not differentially associated with proband status. It was concluded that identifying male multiple-problem youths also identifies families with a high incidence of similar problems. PMID- 9849775 TI - The validity of self-reported cocaine use in a criminal justice treatment sample. AB - Recent studies comparing self-admitted cocaine use with hair and urine test results have raised concerns about underreporting due to variations across situations and settings. Because of the frequent need for self-report data in conducting treatment evaluations, more information is needed on factors that affect the credibility of this information. The present study examines records of cocaine use collected as part of an evaluation of prison-based treatment (N=396). Specifically, self-reported cocaine use from 6-month postrelease follow-up interviews, completed with treatment graduates and a comparison sample of parolees who were eligible but not sent to treatment, was examined in relation to urine and hair test results. Overall, cocaine use was underreported when compared to hair test results, but underreporting was lower for program graduates than for the untreated comparison group. Furthermore, program graduates originally admitted to prison on a drug-related arrest were the most likely to underreport cocaine use, seemingly due to a heightened concern about potential consequences, such as returning to prison on another drug-related charge. Given that the validity of self-report for high-risk individuals varies greatly and is difficult to predict across studies, future research with criminal justice samples should continue to assess the validity of these measures under specific research conditions and for diverse types of individuals. PMID- 9849776 TI - Alcohol availability and domestic violence. AB - The purpose of this study was to examine the relationship among sociodemographic variables, alcohol outlet density, and rate of domestic violence in New Jersey. Data were obtained for the 223 largest municipalities in the state and were examined using factor analysis and bivariate and multivariate analyses. Three sociodemographic factors were extracted through factor analysis. These explained 58% of the variance among municipalities in rates of domestic violence. One factor--termed social disadvantage--explained the greatest amount of unique variance (42%). Alcohol outlet density added nothing to the sociodemographic model and did not interact with any of the three sociodemographic factors. The findings show that, in the state of New Jersey, higher levels of alcohol outlet density are not geographically associated with higher rates of domestic violence. These findings may be due to limitations in the data sets employed in the study, limitations of the macrolevel analysis employed, and/or the complex nature of the relationship between alcohol use and domestic violence. PMID- 9849777 TI - Knowledge and misconceptions among inner-city African-American mothers regarding alcohol and drug use. AB - Alcohol and drug knowledge of inner-city mothers was evaluated following an educational mailing, and the relationship between knowledge and alcohol and illicit drug use during pregnancy was tested. Eighty-four postpartum African American mothers with known alcohol and drug use during pregnancy received a U.S. Department of Education publication, Growing Up Drug Free: A Parent's Guide to Prevention. Results of a phone-administered quiz from this booklet were compiled, and alcohol and drug use subgroups were compared. The average score was 50%. Half of the women did not know that alcohol is the most commonly used drug in the United States. Few identified alcohol, tobacco, and marijuana as the three drugs most commonly used by children. Lack of teenage substance use experience was perceived to increase the risk of chemical dependency. Drinkers and drug users were fourfold more likely to answer at least six questions correctly (p=.03 each, logistic regression). Parental knowledge of substance use, particularly of alcohol, remains inadequate. We suggest that appropriate parental education tools are still needed for optimal primary prevention of substance use by inner-city children. PMID- 9849779 TI - The traditional fermented milk products of the Sudan. PMID- 9849778 TI - MOS-SF-36 in evaluating health-related quality of life in alcohol-dependent patients. AB - Health-related quality of life (HRQoL) was evaluated in a sample of alcohol dependent patients with the 36-item Medical Outcome Study Short-Form Health Survey (MOS-SF-36). The instrument was administered to 147 patients (77% males), aged 26-78, with a DSM-III-R diagnosis of alcohol dependence. The Hamilton Depression Scale (HDS), the Severity of Alcohol Dependence Questionnaire (SADQ), and the Addiction Severity Index (ASI) were also administered to the first 100 patients included in the study. The reliability and validity of the MOS-SF-36 were evaluated. Test-retest intraclass coefficients for a 10-day interval were in the range .65 to .79, whereas the Cronbach alpha coefficient indicated good internal consistency (range .70 to .89). Compared to scores observed in the general population, MOS-SF-36 scores for alcohol-dependent patients were relatively low (indicating worse perception of HRQoL), especially in the psychological and role dimensions (range 52/100 to 55/100), but were closer to populational values in the physical and functional dimensions (range 61/100 to 75/100)). The highest correlation between MOS-SF-36 dimensions and HDS was found in the MOS-SF-36 "mental health" dimension (r=-.56, p < .001); this dimension was also correlated highly with the psychiatric dimension of the ASI (r=-.73, p < .001). The eight dimensions of the MOS-SF-36 were 21% to 127% lower in patients with HDS greater than or equal to 16 (major depression) compared to those with HDS less than or equal to 7 (absence of depression). The MOS-SF-36 dimensions were 10% to 141% lower in patients with high "ASI alcohol" scores, indicating worse HRQoL profiles with a higher severity of alcohol dependence. The MOS-SF-36 presents good criteria for reliability and validity in alcohol-dependent patients. The results suggested that alcohol-dependent patients perceived their problems more as psychological than physical. The severity of alcohol dependence and depression seemed to influence the perception of HRQoL negatively. PMID- 9849780 TI - Effect of enhanced proteolysis on formation of biogenic amines by lactobacilli during Gouda cheese ripening. AB - The effect of enhanced proteolysis on amine formation by amino acid decarboxylase positive Lactobacillus sp. during Gouda cheese ripening was examined. A commercial proteolytic enzyme preparation was added to pasteurized milk prior to cheese preparation. The effect of this manipulation on the formation of putrescine, histamine and tyramine was investigated in the presence and absence of amino acid decarboxylase-positive strains during a 12-week ripening period. Four batches were supplemented with a proteolytic enzyme. Batch I contained the proteolytic enzyme only, whereas batches II-IV were additionally supplemented with Lactobacillus delbrueckii LTH 1260 (batch II), Lactobacillus buchneri LTH 1388 (batch III) or Lactobacillus brevis LTH 2560 (batch IV). In batch I putrescine was detected with 4 mg/kg, in batch II, 42 mg/kg putrescine, 238 mg/kg histamine and 636 mg/kg tyramine were found. Batch III contained 13 mg/kg putrescine and 418 mg/kg histamine, whereas in batch IV, 26 mg/kg putrescine and 776 mg/kg tyramine were present. Batch V was supplemented with all three lactobacilli but did not contain the proteolytic enzyme. In this experiment, 4 mg/kg putrescine, 179 mg/kg histamine and 337 mg/kg tyramine were detected. A control cheese batch (VI) without addition of amine forming lactobacilli or a proteolytic enzyme was produced and only 4 mg/kg putrescine were detected. An increase in amine concentration during cheese ripening under conditions of enhanced proteolysis in the presence of starter and spoilage lactobacilli was evident from the experiments. PMID- 9849781 TI - Predictive model to describe the combined effect of pH and NaCl on apparent heat resistance of Bacillus stearothermophilus. AB - The combined effect of pH and NaCl on the apparent thermal resistance of Bacillus stearothermophilus ATCC 12980 spores was studied. Spores were heated at different temperatures (115-125 degrees C) in mushroom substrate, acidified using glucono delta-lactone to different pH levels (from 5.75 to 6.7), which contained concentrations of NaCl that ranged from 0.5 to 3% (w/v). The recovery medium was acidified to the same pH level and contained the same NaCl concentration as the heating menstruum. A factorial experimental design allowed a predictive model to be developed, which described the combined effect of heating temperature, pH and NaCl on the thermal resistance of B. stearothermophilus spores. Predictions from the model provided a valid description of the data used to generate the model, and agreed with observations from the literature and from an independent experiment performed using asparagus and bean substrates. PMID- 9849782 TI - Bacillus cereus in a whey process. AB - A cheese dairy and its whey manufacturing line were examined for Bacillus cereus. Colonies typical of B. cereus were detected in 120 (17%) samples out of 720 analysed. Only 3% of the sampled raw milk contained B. cereus ( > or = 10 cfu ml( 1)) whereas in evaporated whey concentrate B. cereus was present in 76% of the samples. Nitrate reductase negative and weakly casein hydrolysis isolates were rare in raw milk and the early parts of the process but these defective biotypes became increasingly frequent towards the end of the whey process. The composition of whole cell fatty acids of B. cereus isolates originating from the whey part of the process was different from that of the type strain and of the isolates originating from the raw materials of cheese making. The B. cereus strains in concentrated whey were 100% similar to the type strain in 16S rDNA sequence (500 bp) although they were not or only poorly recognized as B. cereus by a commercial whole cell fatty acid library. All of B. cereus isolates in raw milk were sensitive to one or more of the B. cereus group phages (n = 17) whereas 43% of the isolates from the whey process were sensitive to none. None of the 23 strains originating from the whey processing lines grew at < or = 8 degrees C. although strains with minimum growth temperatures of 5.3 degrees C and 7.0 degrees C were present in the raw materials. Our results indicate that the B. cereus population of the warm ( > 30 degrees C) parts of the cheese dairy process was separate from that of cold (2 degrees C to 4 degrees C) part of the process. PMID- 9849783 TI - Control of the contamination of pig carcasses by Escherichia coli from their mouths. AB - Swab samples from the mouths of 40 pig carcasses all yielded Escherichia coli, at estimated log mean numbers of 2.3 per sample. Single colonies of E. coli O157:H7 were isolated from two, and single colonies of other verotoxigenic strains of E. coli from a further two of those samples. E. coli from the mouth were deposited on the surfaces of carcasses during usual, commercial carcass dressing operations for cutting open the throat and the floor of the mouth, and freeing the tongue. After those operations had been performed, E. coli were recovered from a neck site on 45 of 50 carcasses which were sampled by swabbing that site. When 50 carcasses on which those operations had been performed were pasteurized, the log mean numbers of total aerobic counts recovered from the neck site on carcasses were reduced by two as compared with the unpasteurized carcasses, and only one E. coli was recovered from those carcasses. Pasteurization bleached the small areas of cut muscle exposed during the operations on the mouth and throat, but apparently had no other effect on the appearance of the carcasses. It is suggested that pasteurization of pig carcasses after operating on and regulatory inspection of the mouth and throat may be a commercially viable approach to reducing the contamination of pork with pathogens from the mouths of carcasses. PMID- 9849784 TI - Application of artificial neural networks as a non-linear modular modeling technique to describe bacterial growth in chilled food products. AB - In many chilled, prepared food products, the effects of temperature, pH and %NaCl on microbial activity interact and this should be taken into account. A grey box model for prediction of microbial growth is developed. The time dependence is modeled by a Gompertz model-based, non-linear differential equation. The influence of temperature, pH and %NaCl reflected in the model parameters is described by using low-complexity, black box artificial neural networks (ANN's). The use of this non-linear modeling technique makes it possible to describe more accurately interacting effects of environmental factors when compared with classical predictive microbiology models. When experimental results on the influence of other environmental factors become available, the ANN models can be extended simply by adding more neurons and/or layers. PMID- 9849785 TI - Stability and safety of traditional Greek salami -- a microbiological ecology study. AB - The microbiological and physicochemical changes which occurred during the industrial fermentation and ripening of four batches of Greek dry salami manufactured without starter cultures were followed. Moderated dehydration rates, monitored by slowly decreasing relative humidity from 94 to 90% during fermentation, prevented the production of insufficiently acidified batches by maintaining microbial activity for longer when the natural inoculum was low. The terminal pH values (5.0-5.2) and water contents (27.7-30.3%) of the sausages were narrowly ranged. Fermentation was governed by an active (> 10(8) cfu g(-1)) lactic flora, consisting of 'wild' strains of Lactobacillus sake. Gram-negative bacteria and aerobic sporeformers decreased below 10(2) and l0(3) cfu g(-1), respectively, while yeasts did not significantly increase during ripening and were below 10(5) cfu g(-1) in the ripened product. Sausages were substantially free of sulfite-reducing clostridia and coagulase-positive staphylococci during the whole process. Listeria spp., occurred in the fresh sausage mix, but disappeared from all batches at the latest by the end of fermentation. Enterococci exceeded 10(5) cfu g(-1) during the first days and remained at this level during ripening. Novobiocin-resistant staphylococci matching Staphylococcus saprophyticus (mainly) and S. xylosus dominated Micrococcaceae populations, ranged between 10(5) and 10(6) cfu g(-1). This is the first report of such a large contribution from S. saprophyticus to the production of dry salami of good quality. It is concluded that to keep or improve the traditional 'sensory type' of Greek salamis, suitable strains of L. sake, S. xylosus and possibly nitrate reducing S. saprophyticus should be selected and validated as starter cultures in experimentally inoculated salamis. PMID- 9849786 TI - The effect of the growth environment on the lag phase of Listeria monocytogenes. AB - The duration of lag in Listeria monocytogenes was examined in relation to the physico-chemical properties of the growth environment. It was supposed that lag would be determined by two hypothetical quantities, the amount of work that a cell has to perform to adapt to new conditions and the rate at which it can perform that work. If the rate at which the cell can perform the necessary work is a function of the maximum specific growth rate in the new environment, the hypothesis predicts that lag time should be related in some way to growth rate, provided cells are initially in approximately the same physiological state. Literature data suggest this is true for many organisms when temperature is the sole growth limiting factor. However, lag times of L. monocytogenes displayed an unusual response to temperature in which lag times of cells precultured at 37 degrees C were shorter at 15 degrees C than at 20 degrees C or 25 degrees C. Analysis of data from the Food Micromodel in which growth of L. monocytogenes was controlled by combinations of pH, NaCl concentration and temperature, showed that there was a linear relationship between lag time and mean generation time although there was much scatter in the data. When the effects of pH, solute type and concentration were investigated individually in this work the correlation between lag time and mean generation time was often poor. It would thus appear that the relationship between growth environment and lag time is more complex than the corresponding relationship between growth environment and maximum specific growth rate. PMID- 9849787 TI - Demonstration of safety of probiotics -- a review. AB - Probiotics are commonly defined as viable microorganisms (bacteria or yeasts) that exhibit a beneficial effect on the health of the host when they are ingested. They are used in foods, especially in fermented dairy products, but also in pharmaceutical preparations. The development of new probiotic strains aims at more active beneficial organisms. In the case of novel microorganisms and modified organisms the question of their safety and the risk to benefit ratio have to be assessed. Lactic acid bacteria (LAB) in foods have a long history of safe use. Members of the genera Lactococcus and Lactobacillus are most commonly given generally-recognised-as-safe (GRAS) status whilst members of the genera Streptococcus and Enterococcus and some other genera of LAB contain some opportunistic pathogens. Lactic acid bacteria are intrinsically resistant to many antibiotics. In many cases resistances are not, however, transmissible, and the species are also sensitive to many clinically used antibiotics even in the case of a lactic acid bacteria- associated opportunistic infection. Therefore no particular safety concern is associated with intrinsic type of resistance. Plasmid-associated antibiotic resistance, which occasionally occurs, is another matter because of the possibility of the resistance spreading to other, more harmful species and genera. The transmissible enterococcal resistance against glycopeptide antibiotics (vancomycin and teicoplanin) is particularly noteworthy, as vancomycin is one of the last effective antibiotics left in the treatment of certain multidrug-resistant pathogens. New species and more specific strains of probiotic bacteria are constantly identified. Prior to incorporating new strains into products their efficacy should be carefully assessed, and a case by case evaluation as to whether they share the safety status of traditional food-grade organisms should be made. The current documentation of adverse effects in the literature is reviewed. Future recommendations for the safety of already existing and new probiotics will be given. PMID- 9849788 TI - Adenylates and adenylate-energy charge in submerged and planktonic cultures of Salmonella enteritidis and Salmonella typhimurium. AB - Adenine nucleotide values and adenylate energy charge (AEC) were measured during the growth of Salmonella enteritidis and Salmonella typhimurium as submerged colonies in agarose gel and gelatin gel, and as planktonic cells in broth. Growth in all three systems showed similar trends with a ten-fold decrease in total adenylate pool during exponential growth, before attaining a fairly stable value throughout stationary phase. AEC values were generally low, (approximately 0.66), but did rise slightly during stationary phase. The large proportion of dead cells during early exponential phase may have contributed to the adenosine diphosphate and adenosine monophosphate pools, through cell lysis or excretion, and it is suggested that this was likely to account for the low values of AEC. In agarose and gelatin gelled cultures the percentage of adenosine triphosphate (ATP) in relation to the total adenylates showed random fluctuations. This was contrary to the broth culture where percentage ATP was highest after 12 h and the data formed a smooth curve. These data demonstrated that considerable physiological heterogeneity exists within a colony of bacteria growing in a gel matrix and by analogy a food material also, and that AEC is a poor indicator of cell viability in such systems. PMID- 9849789 TI - Rapid detection of Escherichia coli O157:H7 using immuno-magnetic flow cytometry in ground beef, apple juice, and milk. AB - A rapid method combining flow cytometry and immunomagnetic bead separation (IMFC) was effective for detecting low populations of Escherichia coli O157:H7 inoculated into ground beef, apple juice, and raw milk samples. Modified buffered peptone was most effective for enrichment of E. coli O157:H7 in ground beef. Using IMFC in combination with a 6-h enrichment, as few as four E. coli O157:H7 cells/g of ground beef were detected in as little as a 7-h total analysis time. Our results demonstrate the possibility of using flow cytometry and immunomagnetic bead separation to detect low concentrations of E. coli O157:H7 in ground beef, apple juice and milk. PMID- 9849790 TI - Influence of several environmental factors on the initiation of germination and inactivation of Bacillus cereus by high hydrostatic pressure. AB - The influence of pH, aw, L-alanine, and fat concentration of milk on the initiation of germination and inactivation by high hydrostatic pressure (HHP) (250 mPa at 25 degrees C for 15 min and 690 mPa at 40 degrees C for 2 min) of Bacillus cereus sporulated at 20, 30 and 37 degrees C was investigated. B. cereus sporulated at the lowest temperature was found to be the most resistant to the initiation of germination and inactivation by HHP. At ambient pressure, the rate and extension of germination induced by L-alanine were also lower in B. cereus sporulated at 20 than 30 or 37 degrees C. The optimum pH for the germination and inactivation of B. cereus depended on the sporulation temperature. At 250 mPa the extent of germination for the three suspensions increased with higher pH. At 690 mPa, the pH barely affected the germination of B. cereus sporulated at 20 degrees C (3 log cycles), but the inactivation increased as the pH of the medium was lowered. After the same treatment, pressure optimally initiated the germination of B. cereus sporulated at 30 and 37 degrees C (6-7 log cycles) around neutral pH. Higher inactivation was obtained at pH 6. High concentrations of sucrose protected the spores from the germinating and inactivating effect of HHP. At aw 0.92, no germination was detected when the spores were pressurized at 250 mPa, and only 1 log cycle of B. cereus sporulated at 20 and 30 degrees C and 2 log cycles of B. cereus sporulated at 37 degrees C were germinated at 690 mPa. In addition, no inactivation was observed at aW 0.92 even at 690 mPa. The presence of L-alanine in the medium of pressurization increased the germination initiated by HHP at 250 mPa, but not at 690 mPa. A combination of 250 mPa at 25 degrees C with L-alanine (100 mM) was found to give an additive response. The initiation of germination and inactivation by HHP were not affected by the fat concentration of the milk. PMID- 9849792 TI - Initial numbers, serovars and phagevars of Listeria monocytogenes isolated in prepared foods in the city of Barcelona (Spain). AB - A total of 1100 samples of prepared foods purchased in restaurants and delicatessen shops in the city of Barcelona was examined for the presence of Listerio. L. monocytogenes was more frequently isolated from foods intended to be consumed without further cooking (9.3%) than from foods intended to receive further cooking prior to consumption (2.9%). A quantitative study, carried out with 773 samples, yielded 1% of samples with numbers of L. monocytogenes higher than 100 CFU/g. Strains of L. monocytogenes belonged to serovar 4b (36.3%), 1/2a (29.5%), 1/2b (22.7%) and 1/2c (11.3%). L. monocytogenes serovar 4b phagevar 3274: 2671 (6 of 11) was prevalent among the strains studied. PMID- 9849791 TI - Effect of water activity and temperature on growth and ochratoxin production by three strains of Aspergillus ochraceus on a barley extract medium and on barley grains. AB - Aspergillus ochraceus Wilhelm is a widespread storage fungus that has been isolated from grains such as barley. The objective of this study was to determine the effects of water activity (a(w)), temperature, time and their interactions on (a) growth on a barley extract agar medium, (b) growth on barley grains and (c) ochratoxin production on barley grains by three strains of A. ochracieus. For the three A. ochreceus isolates examined (NRRL 3174, 3.113 and 3.38), optimal a(w) levels for growth on agar media were in the range 0.98-0.96, with temperature optima of 30 degrees C for two of the isolates and 25-30 degrees C for the other isolate. Growth was observed at 10 and 37 degrees C, but only at higher a(w) levels assayed. Two dimensional profiles were constructed for the range of a(w) and temperature conditions that allow growth of the three isolates. Maximum growth on barley grains was reached at 30 degrees C, at both a(w) levels assayed (0.96 and 0.98), with fungal growth rates in the 4-5 mm day(-1) range. Maximum amounts of ochratoxins were produced at the highest a(w) treatment (0.98 a(w)) and after a three-week incubation time at 25-30 degrees C. The range of ochratoxin concentrations varied considerably, from 1.7 to 12,949 ppm, depending on the a(w) and temperature interactions. PMID- 9849793 TI - Linomide suppresses chronic-relapsing experimental autoimmune encephalomyelitis in DA rats. AB - Linomide (quinoline-3-carboxamide) is a synthetic immunomodulator that suppresses several experimental autoimmune diseases. Here we report the effects of Linomide on chronic progressive and/or relapsing experimental autoimmune encephalomyelitis (PR-EAE), a CD4+ T cell mediated animal model of multiple sclerosis (MS). PR-EAE induced in DA rats by inoculation with homogenized guinea pig spinal cord and Freund's complete adjuvant, was strongly suppressed by Linomide administered daily subcutaneously from the day of inoculation. Linomide dose-dependently delayed the interval between immunization and onset of clinical PR-EAE, reduced severity and relapse of clinical PR-EAE, and shortened clinical PR-EAE. These clinical effects were associated with the down-modulation of CNS antigen-induced T cell responses and production of proinflammatory cytokines (IFN-gamma and TNF alpha) as well as with upregulation of IL-4 (except in spleen MNC), IL-10 and TGF beta in both spleen MNC and the spinal cord. These effects indicate that Linomide can suppress PR-EAE and may mediate its suppressive effects by regulation of cytokines. PMID- 9849794 TI - Sequential changes of dopaminergic receptors in the rat brain after 6 hydroxydopamine lesions of the medial forebrain bundle. AB - We investigated the sequential patterns of changes in dopamine uptake sites, D1 and D2 receptors in the brain of animals lesioned with 6-hydroxydopamine using quantitative receptor autoradiography. The rats were unilaterally lesioned in the medial forebrain bundle and the brains were analyzed at 1, 2, 4 and 8 weeks postlesion. Degeneration of the nigrostriatal pathway caused a significant loss of dopamine uptake sites in the ipsilateral striatum, substantia nigra (SN) and ventral tegmental area (VTA) in the lesioned animals. Dopamine D1 receptors were significantly increased in the ventromedial part of striatum of the ipsilateral side from 2 to 4 weeks postlesion. In the ipsilateral SN, a transient increase in dopamine D1 receptors was observed only 1 week after lesioning. However, the frontal cortex, parietal cortex and dorsolateral part of the striatum showed no significant change in dopamine D1 receptors throughout the experiments. On the other hand, dopamine D2 receptors were decreased increased in the ipsilateral SN and VTA from 1 week to 8 weeks postlesion. In the ipsilateral striatum, dopamine D2 receptors were increased in the dorsolateral part from 2 weeks to 8 weeks and in the ventromedial part from 2 weeks to 4 weeks. However, the frontal cortex and parietal cortex showed no significant change in dopamine D2 receptors during postlesion. In the contralateral side, most of regions examined showed no significant change in dopamine uptake sites, dopamine D1 receptors and dopamine D2 receptors during postlesion except for a transient change in a few regions. These results demonstrate that 6-hydroxydopamine can cause a severe functional damage in dopamine uptake sites in the striatum, SN and VTA. Our findings also suggest that the up-regulation in dopamine D2 receptors is more pronounced than that in dopamine D1 receptors in the brain after 6-hydroxydopamine treatment. Furthermore, our results support the existence of dopamine D2 receptors on the neurons of SN and VTA. Thus, our findings provide insights into the pathogenesis of Parkinson's disease. PMID- 9849795 TI - Creutzfeldt-Jakob disease via dural and corneal transplants. AB - A review of all published cases of iatrogenic Creutzfeldt-Jakob disease (CJD) via dural (N=71) and corneal (N=4) transplants is given. All but three of the dural cases were obviously due to a commercial product recalled in 1996. Two of the corneal grafts were taken from patients who had died of sporadic CJD. These cases differed from CJD due to human growth hormone injections and the new variant. Instead. they were akin to sporadic cases, but memory loss, disorders of higher cerebral functions and extrapyramidal signs were fewer, while cerebellar abnormalities were more frequent. Progressive dysarthria and gait disorder/gait ataxia were prominent signs during the early stages, myocloni the most salient feature later. A nonperiodic EEG did not contradict the diagnosis. Using current diagnostic criteria the disease was underdiagnosed ante mortem. Utmost care is needed in selecting, harvesting and handling dural and corneal grafts to avoid inadvertent transmission of CJD. PMID- 9849796 TI - Collagen abnormalities in the spinal cord from patients with amyotrophic lateral sclerosis. AB - During the last 10 years, we have demonstrated morphological and biochemical abnormalities of skin extracellular matrices in amyotrophic lateral sclerosis (ALS). However, currently little is known concerning collagen of the spinal cord in ALS. We measured the amount of collagen and characterized collagen at light and electron microscopic levels in posterior funiculus, posterior half of lateral funiculus and anterior horn of cervical enlargement of the spinal cord obtained from ten patients with ALS, 11 patients with other neurologic diseases (control group A), and ten patients without neurologic ones (control group B). In posterior half of lateral funiculus and anterior horn, (1) by light microscopy, there was no significant difference in vessel wall area between ALS patients and control groups A and B; (2) ultrastructurally, collagen bundles were more fragmented and widely separated, and the fibrils were randomly oriented in the perivascular space of capillaries in ALS patients, which were not observed in any areas of control groups or in posterior funiculus of ALS patients; and (3) the collagen contents in ALS were significantly lower (P<0.001 and P<0.001, respectively) than those in control groups A and B. Fragmented and widely separated collagen bundles in the interstitial tissue surrounding capillaries and markedly decreased amount of collagen in posterior half of lateral funiculus and in anterior horn of ALS could be related to the degeneration of the upper and lower motor neurons in the spinal cord in ALS, that is, selective neuronal vulnerability in ALS. PMID- 9849797 TI - Brain biochemistry in Duchenne muscular dystrophy: a 1H magnetic resonance and neuropsychological study. AB - Duchenne muscular dystrophy (DMD) is a progressive muscle disorder associated with an intellectual deficit which is non-progressive. We obtained localised 1H magnetic resonance spectra from the left frontal lobe and left cerebellum of 15 boys with DMD (mean age 106 months+/-32) and 15 similarly aged control boys (mean age 115 months+/-31); all boys underwent a battery of neuropsychological tests. We found a significant (P<0.01) increase in the ratio of choline-containing compounds to N-acetylaspartate (Cho/NA) in the left cerebellum in boys with DMD compared with control boys. There was no change in the creatine/NA ratio and a significant increase (P=0.03) in the Cho/creatine ratio, suggesting that the change in Cho/NA ratio was due to an increase in choline-containing compounds; this increase has been previously observed in the brain of the murine model of DMD, the mdx mouse. No significant changes were observed in spectra obtained from left frontal lobe in DMD compared to controls. We also observed a significant association between Cho/NA in the left cerebellum, and the performance of DMD boys on the Matrix Analogies Test (MAT). The MAT is a test of visuo-spatial ability and non-verbal reasoning which requires neither manual dexterity nor a verbal response for an adequate performance. A comparison of DMD boys whose cerebellar Cho/NA fell within 2 standard deviations of the control norm (0.56+/ 0.24) with DMD boys whose cerebellar Cho/NA was outside this range (i.e. >0.80) revealed a significant difference in ability on the MAT (P<0.05). DMD boys whose Cho/NA ratio is more than two standard deviations higher than controls perform significantly better on the MAT than DMD boys whose Cho/NA ratio is within the normal range. This finding suggests that the observed elevation in Cho/NA and Cho/creatine is not associated with intellectual deficit (as sampled by the MAT), and may represent a compensatory mechanism. The possible interpretations of these metabolic changes are discussed. PMID- 9849798 TI - Coagulation abnormalities in adults with cryptogenic stroke and patent foramen ovale. AB - A patent foramen ovale (PFO) is found by transesophageal echocardiography in one half of patients with cryptogenic stroke (CS). Coagulation abnormalities may promote paradoxical emboli in these patients. Seventeen patients were identified with PFO and CS. Thirty-one percent of patients had hematologic risk factors for venous thrombosis. These included abnormal activated protein C resistance and increased anticardiolipin antibodies. Patients with coagulation abnormalities and a PFO were three times as likely to be treated with warfarin compared to aspirin (P<0.05). Prothrombotic states are common in patients with PFO and CS and identifying these conditions may impact the choice of antithrombotic therapy. PMID- 9849799 TI - Downbeat nystagmus in two siblings with spinocerebellar ataxia type 6 (SCA 6). AB - We report two siblings with spinocerebellar ataxia type 6 (SCA 6), both showing downbeat nystagmus (DBN) as a predominant clinical feature. Familial hemiplegic migraine (FHM), episodic ataxia type 2 (EA-2) and SCA 6 are allelic disorders, and interestingly, the occasional presence of DBN in EA-2 was reported. Our observations suggest that common molecular mechanisms might underlie DBN in FHM, EA-2 and SCA 6. Then, these disorders should be kept in mind in diagnosing patients with DBN. PMID- 9849800 TI - Fatigue in multiple sclerosis: interrelations between fatigue complaints, cerebral MRI abnormalities and neurological disability. AB - Although fatigue is a frequent complaint of patients with multiple sclerosis (MS), little is known about the origins of multiple-sclerosis-associated fatigue. Our primary focus was to study if the extent of cerebral abnormalities, as shown on magnetic resonance imaging (MRI), had any relation with the frequency and intensity of fatigue complaints of patients with a definite diagnosis of MS. Fatigue severity was rated by the patients with the use of a 2-week diary and a fatigue questionnaire, while conventional T1- and T2-weighted MRI provided several measures for cerebral abnormalities. In total, 72% of 45 patients reported to be seriously fatigued at least several times a week over the last 3 month period. Fatigue severity was not related to the total extent of cerebral abnormalities, or to MRI-based atrophy measures. Regional lesion load did not differ between fatigued and non-fatigued subjects. Although neurological disability, as measured by the Expanded Disability Status Scale (EDSS) and Neurological Rating Scale (NRS), did correlate significantly with most MRI measures, it showed no relation with fatigue severity. Neurological progression rates and number of exacerbations in the 2-year period prior to assessment were not significantly associated with the fatigue measures. Therefore, our findings suggest that differences in levels of self-reported fatigue in patients with multiple sclerosis cannot merely be explained by the degree of clinical disease activity, neurological disability or the extent of MRI abnormalities. These results are compared to other research findings and the possible role of alternative factors influencing fatigue in multiple sclerosis are discussed. PMID- 9849801 TI - Risk factors for idiopathic cerebellar ataxia of late onset. AB - In an attempt to identify risk factors for the development of idiopathic cerebellar ataxia (IDCA) we performed a case-control study of 59 IDCA patients. Hypertension and medicine intake were less frequent in IDCA than in neurological controls. Multiple logistic regression yielded an odds ratio (OR) for hypertension of 0.13 (95% confidence interval: 0.00-1.02, P=0.0527) and medicine intake of 0.10 (95% confidence interval: 0.00-0.72, P=0.0157). In contrast, we did not identify an association of IDCA with a number of medical diseases, head trauma, smoking, alcohol intake, rural living and well-water drinking. Some of these factors have been previously shown to be associated with other neurodegenerative diseases. In addition, serum antibody titers against neurotropic viruses were not elevated in IDCA. PMID- 9849802 TI - Lysosomal glycogen storage disease with normal acid maltase with early fatal outcome. AB - In a male infant who had cardiomyopathy, generalized muscle weakness and increased serum creatine kinase levels, his muscle biopsy revealed myopathic changes with tiny intracytoplasmic vacuoles containing PAS-positive material and high acid phosphatase activity, but had normal acid maltase activity biochemically. These findings were consistent with those seen in lysosomal glycogen storage disease with normal acid maltase (Danon disease). Sarcolemmal indentations commonly seen in this disease were missing, but a complement membrane attack complex, C5b-9 was positive along the surface membrane of the muscle fibers as seen in X-linked vacuolar myopathy. The patient was on a respirator and died at 27 months of age from pneumonia and hypertrophic cardiomyopathy. Lysosomal glycogen storage disease with normal acid maltase may be manifested at birth with marked skeletal and cardiac involvement leading to death in early infancy. PMID- 9849803 TI - Bilateral borderzone brain infarctions in association with heroin abuse. AB - A 25-year-old drug abuser who developed an unusual pattern of cerebral ischemic lesions is presented. Cerebral magnetic resonance imaging revealed bilateral borderzone infarctions which were attributed to a heroin-associated vasculitis of the basal cerebral arteries. Under probatory corticosteroid medication the mild neurological deficits completely disappeared. PMID- 9849804 TI - Leber's Hereditary Optic Neuropathy (LHON) with 14484/ND6 mutation in a North African patient. AB - We report the clinical and genetic study of a Leber's Hereditary Optic Neuropathy (LHON) patient of North African origin harboring the 14484/ND6 mutation of mtDNA. For over a year we followed the ophthalmological course of this 24-year-old male with LHON treated with idebenone and vitamin B12. Serum lactate after effort was evaluated before, during and after therapy. Muscle biopsy was obtained for morphological study. Homo/heteroplasmy of 14484/ND6 mutation was studied in different tissues. Recovery of visual acuity was documented 6 months after onset and 3 months after therapy was established. Baseline serum lactate was elevated but normalized after 3.5 months of therapy. Muscle biopsy demonstrated only a few fibers with a slightly increased subsarcolemmal SDH activity. Genetic analysis showed homoplasmic 14484/ND6 mutation in all tissues investigated. The clinical phenotype of LHON/14484 in this patient closely resembles that commonly found in European patients. Even if LHON/14484 patients are reported to have a better prognosis for visual recovery, it is possible that the evolution of visual recovery in this patient could have been influenced by therapy as suggested by changes in serum lactate levels. Bioenergetic impairment of skeletal muscle was documented by lactate levels and muscle morphology. The 14484/ND6 mutation behaves as a primary mutation regardless of mtDNA population-specific backgrounds. PMID- 9849805 TI - Non-familial olivopontocerebellar atrophy combined with Alzheimer's disease. PMID- 9849806 TI - Positron emission tomography in vascular dementia. PMID- 9849807 TI - Activin: an endocrine or paracrine agent? PMID- 9849808 TI - A case for glucosamine. PMID- 9849809 TI - Growth hormone for the failing heart. PMID- 9849810 TI - Glucocorticoid-stimulated gene expression knocked out by knock-in mutation. PMID- 9849811 TI - Hormone delivery: small synthetic molecular mimics. PMID- 9849812 TI - Influence of age and sex on serum concentrations of total dimeric activin A. AB - Several studies have shown that activin A is secreted in substantial amounts into the systemic circulation. The changes that occur during menstrual cycle and pregnancy suggest a correlation with reproductive function. At present, however, no definitive evidence has confirmed this pattern throughout adult life; moreover, neither the origin nor the physiological implications of this circulating growth factor have been clearly defined. The aim of the present study was to evaluate whether circulating concentrations of activin A change in adult men and women according to age and sex, and to examine the possible correlation with serum concentrations of FSH. Total dimeric activin A was measured using a specific two-site enzyme immunoassay in serum specimens collected from a cohort of normal individuals enrolled in an epidemiological survey. A group of men (n = 106) and one of women (n=151) were subdivided into six age groups (20-30, 30-40, 40-50, 50-60, 60-70 and 70-90 years). In a small group of 8 men and 11 women, serum concentrations of activin A were evaluated twice, in specimens collected at an interval of 10 years. Serum FSH concentrations were also measured in all specimens. Serum concentrations of activin A were not significantly different in men and women and showed an age-related progressive increase between 20 and 50 years of age (P<0.01, those aged 40-50 compared with those aged 20-30 years). After the age of 50 years, activin A concentrations remained in the same range of values in women, whereas they increased significantly in men, reaching peak values between 70 and 90 years (P<0.01 compared with the group aged between 20 and 50 years). From the age of 50 years, activin A concentrations were significantly greater in men compared with those in women in the corresponding age groups (P<0.001). Activin A concentrations correlated with age in men, but not in women. No significant correlation between concentrations of activin A and FSH was found in either sex. Activin A concentrations in specimens collected 10 years apart showed an increase in seven of eight men, but not in women. Finally, no significant variations of activin A concentrations were observed when fertile and postmenopausal women were compared. The present data indicate that circulating concentrations of activin A vary according to age; furthermore, men older than 50 years have greater concentrations than women. These changes, which occur irrespectively of FSH concentrations, indicate that circulating activin A is not a hormone of the reproductive axis. PMID- 9849813 TI - L-thyroxine treatment and neurotransmitter levels in the cerebrospinal fluid of hypothyroid patients: a pilot study. AB - Monoamine precursors, neurotransmitters and their metabolites were studied in cerebrospinal fluid (CSF) obtained from nine newly diagnosed hypothyroid patients. Before treatment, the serum TSH correlated positively with the CSF concentrations of tyrosine and phenylalanine. During treatment, the levels of the precursors tryptophan, phenylalanine and tyrosine decreased significantly, as was also the case with dopamine and the noradrenaline metabolite 4-hydroxy-3 methoxyphenylglycol (HMPG), but not with serotonin, noradrenaline and the serotonin metabolite 5-hydroxyindoleacetic acid, nor the dopamine metabolites homovanilic acid and dihydroxyphenylacetic acid. The study provided some indication that the CSF levels of phenylalanine and tyrosine are related to thyroid function. Furthermore, we have found an indication that L-thyroxine treatment affects the CSF levels of the precursors as well as dopamine and HMPG. Our results support the notion that there is an interaction between thyroid function and CSF disposition of monoamine compounds. PMID- 9849814 TI - Analyses of the molecular forms of serum thyroglobulin from patients with Graves' disease, subacute thyroiditis or differentiated thyroid cancer by velocity sedimentation on sucrose gradient and Western blot. AB - Serum thyroglobulin (Tg) concentration increases in diverse thyroid pathophysiological situations. We examined whether Tg molecules appearing in the serum of patients with Graves' disease (GD), subacute thyroiditis (ST) or differentiated thyroid cancer (DTC) have distinctive biochemical properties. We used ultracentrifugation on sucrose gradient and Western blot to analyze structural parameters of immunoreactive Tg in complete serum from 40 patients with GD, ST or DTC. Purified human Tg was used as reference. Immunoreactive Tg from ST or DTC sera sedimented in a single, rather symmetrical peak as purified 19S Tg. In GD sera without detectable anti-Tg autoantibodies (TgaAb), about 80% of immunoreactive Tg was recovered in a Tg dimer peak that often split into two components; the remaining Tg immunoreactivity (10-30%) sedimented faster and was polydispersed. In GD sera containing TgaAb, immunoreactive Tg was recovered in a peak sedimenting faster than 19S Tg corresponding to immune complexes identified by protein A adsorption. Using a Western blot procedure, optimized to detect 0.1 ng Tg in serum. a single band of Tg, migrating as the intact Tg subunit, was always found in non-reducing conditions; the intensity of the band correlated with the immunoassayable Tg concentration. In reducing conditions, the Tg band obtained with GD or ST sera was decreased by up to 70% compared with that of purified Tg or serum Tg from patients with DTC. In conclusion, serum Tg from DTC is remarkably homogeneous and in the form of dimers dissociable into uncleaved monomers. In contrast, serum Tg from GD or ST is heterogeneous with respect to its sedimentation properties and/or the structural integrity of its polypeptide chains. These data provide information on the processes whereby Tg is released into the circulation. PMID- 9849815 TI - Thyroxine administration to infants of less than 30 weeks gestational age decreases plasma tri-iodothyronine concentrations. AB - OBJECTIVE: To investigate the effect on thyroid hormone metabolism of the administration of thyroxine to very preterm infants. DESIGN AND METHODS: Two hundred infants of less than 30 weeks gestation were enrolled into a randomized, double-blind, placebo-controlled trial. Thyroxine (T4) (at a fixed daily dose of 8 microg/kg birthweight) or placebo was started 12-24h after birth and discontinued 6 weeks later. Plasma concentrations of T4, tri-iodothyronine (T3), reverse T3 (rT3), TSH, and thyroxine-binding globulin were measured weekly during trial medication and 2 weeks thereafter. RESULTS: The T4 and the placebo group each comprised 100 infants. Antenatal, perinatal, and postnatal clinical characteristics were comparable in both groups. T4 and rT3 were significantly increased in the T4 group. TSH concentrations were depressed in the T4 group and T3 was significantly decreased, probably as a result of TSH depression. The T4/T3 and T4/rT3 ratios differed significantly between the two study groups. CONCLUSIONS: Daily T4 administration during the first 6 weeks after birth to infants of less than 30 weeks gestation prevents hypothyroxinemia, but decreases plasma T3 concentrations. Our finding possibly implies that very preterm infants should receive supplements of both T4 and T3. PMID- 9849816 TI - Cabergoline in acromegaly: a renewed role for dopamine agonist treatment? AB - OBJECTIVE AND DESIGN: Eighteen active acromegalics entered a prospective open study with cabergoline (CAB), a dopaminergic drug much more potent than bromocriptine (Br). METHODS: CAB was administered for 6 months at doses ranging between 0.5 mg twice weekly and 0.5 mg/day. Clinical-anamnestic characteristics of the patients were: (i) sensitivity to dopamine agonist drugs (10 patients); (ii) resistance to somatostatin analogs (SAs) (8 patients): (iii) intolerance to SA (3 patients). In 2 patients marked hyperprolactinemia was present. RESULTS: Basal GH was 6.6 microg/l (2.2-50) (median (range)), and on treatment it was 3.5 microg/l (1.2-34) (P=0.013). The corresponding IGF-I values were 720 microg/l (410-1438) and 375 microg/l (167-1260) respectively (P=0.00001). Individual GH levels decreased below 2 microg/l in 5 patients, and between 2 and 5 microg/l in another 5 patients. IGF-I levels were suppressed below 50% of baseline in 8 patients and normal age-adjusted IGF-I values were reached in 5 patients (27% of the series). The retrospective comparison with previous chronic treatment with Br in the 10 suitable patients showed a greater effectiveness of CAB (IGF-I decrease on CAB treatment, 46.8%, on Br treatment, 31%, P=0.02). Adenoma shrank in the 3 patients whose pituitary imaging was repeated during CAB. CONCLUSIONS: These results envisage that CAB may represent a worthy therapeutic tool in acromegalic patients, inducing a degree of IGF-I and GH suppression comparable to SAs, administered by the oral route and much less expensive. PMID- 9849817 TI - Advanced onset of menarche and higher bone mineral density depending on vitamin D receptor gene polymorphism. AB - Bone mineral density (BMD) at distal forearm, and weight and height of healthy Japanese girls aged 18-19 years were measured and their age at menarche was obtained through a questionnaire. A statistically significant association was found between BMD at distal radius and vitamin D receptor (VDR) gene polymorphism at the ApaI site. The age at menarche in the population with Aa genotype was significantly earlier than that in the aa population. In addition, BMD was significantly dependent on the earlier onset of menarche in the population with genotype Aa but not in the population with genotype aa. BMD was also positively associated with the body mass index (BMI) in the population with genotype Aa. Statistical analysis suggested a stronger effect of VDR genotype on age at menarche than on BMI. Thus, we show that VDR gene polymorphism advances the age at menarche and increases BMD in cooperation with age at menarche. PMID- 9849818 TI - Changes in plasma leptin during the menstrual cycle. AB - OBJECTIVE: To measure the plasma concentration of leptin, which is expressed in ovarian follicles and may have a reproductive function, in healthy women during the menstrual cycle. DESIGN: This study included nine women with regular menstrual cycles (mean+/-S.E.M. age 28+/-2 years: body mass index 23.9+/-1.8 kg/m2). From the onset of menses, fasting blood samples were collected every 1-2 days throughout the menstrual cycle. As a control, plasma leptin was measured in six postmenopausal women and six men every other day for 28 days. RESULTS: In menstruating women, plasma leptin increased from 14.9+/-2.9 ng/ml in the early follicular phase to 20.4+/-4.2 ng/ml (P<0.01) at the midluteal phase and returned to the baseline by the subsequent menses. In contrast, leptin concentrations did not change significantly in postmenopausal women or men. The changes in plasma leptin during the menstrual cycle were not related to changes in sex hormones. CONCLUSIONS: The cause of the increase in plasma leptin during the late follicular and luteal phases of the menstrual cycle is not clear. It may be attributed to augmented adipocyte production of leptin in response to increased caloric intake or hypothalamic release of neuropeptide Y. or to release of leptin from mature ovarian follicles. Leptin may have a role in regulating the menstrual cycle and preparing the body for the metabolic demands of pregnancy. PMID- 9849819 TI - Dose-related effects of growth hormone on IGF-I and IGF-binding protein-3 levels in non-islet cell tumour hypoglycaemia. AB - Mesenchymal tumours are a well recognised cause of spontaneous hypoglycaemia. The mechanism is thought to relate to hypersecretion by tumour cells of high molecular mass insulin-like growth factor-II (pro-IGF-II), with consequent suppression of growth hormone (GH) secretion. The use of recombinant human (rh)GH has been reported to alleviate hypoglycaemia in non-islet cell tumour hypoglycaemia, and the mechanism is thought to relate to GH-mediated increments in serum levels of IGF-binding protein-3 (IGFBP-3), thereby reducing the bioavailability of IGF-II. We report the effect of increasing doses of rhGH on the clinical condition and serum IGF-I and IGFBP-3 levels in two patients with solitary pleural fibrous tumours causing severe hypoglycaemia. Hypoglycaemia was successfully alleviated in each patient although, despite using large doses of rhGH, the observed increments in IGFBP-3 were only modest. We postulate that the beneficial effects of rhGH in this situation are likely to be multifactorial and not simply related to increments in serum IGFBP-3 levels. PMID- 9849820 TI - Increased expression of tumor necrosis factor-alpha and decreased expression of thyroglobulin and thyroid peroxidase mRNA levels in the thyroids of iodide treated BB/Wor rats. AB - Several lines of evidence suggest that tumor necrosis factor-alpha (TNFalpha) may contribute to the pathogenesis of autoimmune thyroid disease. It is not known, however, whether increased thyroidal TNFalpha levels are associated with changes in thyroid function. The purpose of the present study was to utilize in situ hybridization histochemistry and immunohistochemistry to determine if the expression of TNF-alpha in the thyroid is associated with a decrease in thyroglobulin (Tg) and thyroid peroxidase (TPO) mRNA levels. Lymphocytic thyroiditis was induced in BB/Wor rats by iodide administration, and thyroidal Tg and TPO mRNA levels were assessed by Northern blot analysis and in situ hybridization, and TNFalpha expression by Northern blot analysis and immunohistochemistry. Thyroids were obtained before and 1 and 2 months after iodide administration. Hematoxylin and eosin staining revealed that there was a progressive increase in mononuclear cells in the thyroids of BB/Wor rats ingesting iodide for 1 and 2 months. Northern blot analysis revealed that during the same time course there was a progressive increase in TNFalpha mRNA levels and a progressive decrease in Tg and TPO mRNA levels in the thyroids. In situ hybridization histochemistry was performed to determine if the decrease in Tg and TPO mRNA levels was associated with thyroid follicular cells in contact with infiltrating mononuclear cells. In rats treated with iodide for 1 month, there was a modest decrease in Tg and TPO mRNA levels in follicular cells in contact with infiltrating mononuclear cells. After 2 months of iodide treatment there was clearly a localized decrease in Tg and TPO mRNA levels in follicular cells in contact with infiltrating mononuclear cells. Immunohistochemical analysis did not detect TNFalpha in the thyroids from control rats or from rats treated with iodide for 1 month. In contrast, after 2 months of treatment, TNFalpha was easily detected in infiltrating mononuclear cells and in some thyroid follicular cells. Together, these results suggest that the suppression of Tg and TPO mRNA levels was associated with the expression of TNFalpha and thus are in agreement with in vitro studies demonstrating that TNFalpha inhibits thyroid cell function. PMID- 9849821 TI - Effects of DPDPE (a specific delta-opioid receptor agonist) and naloxone on hypothalamic monoamine concentrations during the pre-ovulatory LH surge in the rat. AB - We have investigated the inter-relationship between the opioid and aminergic systems in the control of secretion of the pro-oestrous LH surge and the involvement of delta-opioid receptor subtypes in this process. Conscious female rats bearing a cannula in the femoral artery were injected i.p. with a selective delta-opioid receptor agonist (DPDPE) either alone or with the opioid antagonist (naloxone) at 1300 h on the day of pro-oestrus. Blood samples were collected hourly between 1500 h and 1900 h, and plasma LH levels were measured by RIA. At the end of this period (1900 h), the animals were autopsied and the concentrations of the amines (noradrenaline (NA), dopamine (DA), 5 hydroxytryptamine (5HT)) and their metabolites (dihydroxyphenolglycol (DHPG) and 5-hydroxyindoleacetic acid (5HIAA), metabolites of NA and 5HT respectively) were determined by HPLC with electrochemical detection in the medial preoptic area, suprachiasmatic nucleus, median eminence and arcuate nucleus. DPDPE abolished the LH surge and concomitantly decreased hypothalamic NA and DHPG concentrations in all the areas examined. The levels of DA, 5HT and 5HIAA were also reduced in all hypothalamic regions studied, except DA and 5HIAA in the suprachiasmatic nucleus. Naloxone reversed these inhibitory effects of the delta-agonist. We conclude that activation of delta-opioid receptors may exert an inhibitory effect on LH release. The effect is probably an indirect one mediated by the monoaminergic systems, as they are suppressed by DPDPE in nearly all the hypothalamic regions studied. PMID- 9849823 TI - Trends in HIV seroprevalence, AIDS and prevention policy among intravenous drug users and men who have sex with men, before and after 1990 in Austria. AB - This study reports for the first time on secular trends in human immunodeficiency virus (HIV) infection and AIDS, and possible associations with prevention policy in Austria. We analysed HIV seroprevalence and AIDS cases among intravenous drug users (IDU) and men who have sex with men (MSM). In this study we found a diminished rate of increase in new cases of AIDS and a decline in HIV seroprevalence among IDU but not among MSM. Among clients visiting HIV counselling and testing centres in Austria between 1987 and 1990, seroprevalence among IDU was estimated at 27.9% as compared to 19.6% between 1990 and 1992 (odds ratio (OR): 0.62; 0.45-0.85). Among MSM corresponding prevalence for these two periods was 12.1% and 10.9%, respectively, which was not a significant decline. In the period 1990 to 1994, the increase in AIDS cases per half-year levelled off for IDU (incidence rate ratio (IRR) :1.00; 0.99-1.01) but to a lesser extent among MSM (IRR: 1.01; 1.01-1.02). The most effective prevention policy intervention was considered to be the national Methadone Maintenance Program (MMTP), started in 1987, and the provision of sterile injection equipment. We observed that in the recent period there was a decline in the frequency of attendance among young (less than 28 years of age) MSM at counselling centres (OR: 1.27; 95 % CI: 1.08-1.49), accompanied by the observation that the rate of seroprevalence among this group did not decline. This is in contrast to young IDU where attendance did not decline but seroprevalence did. Although inference is limited from cross sectional studies, we argue for a reoriented and effectively monitored HIV prevention policy focused on young MSM. PMID- 9849822 TI - Ipamorelin, the first selective growth hormone secretagogue. AB - The development and pharmacology of a new potent growth hormone (GH) secretagogue, ipamorelin, is described. Ipamorelin is a pentapeptide (Aib-His-D-2 Nal-D-Phe-Lys-NH2), which displays high GH releasing potency and efficacy in vitro and in vivo. As an outcome of a major chemistry programme, ipamorelin was identified within a series of compounds lacking the central dipeptide Ala-Trp of growth hormone-releasing peptide (GHRP)-1. In vitro, ipamorelin released GH from primary rat pituitary cells with a potency and efficacy similar to GHRP-6 (ECs) = 1.3+/-0.4nmol/l and Emax = 85+/-5% vs 2.2+/-0.3nmol/l and 100%). A pharmacological profiling using GHRP and growth hormone-releasing hormone (GHRH) antagonists clearly demonstrated that ipamorelin, like GHRP-6, stimulates GH release via a GHRP-like receptor. In pentobarbital anaesthetised rats, ipamorelin released GH with a potency and efficacy comparable to GHRP-6 (ED50 = 80+/ 42nmol/kg and Emax = 1545+/-250ng GH/ml vs 115+/-36nmol/kg and 1167+/-120ng GH/ml). In conscious swine, ipamorelin released GH with an ED50 = 2.3+/-0.03 nmol/kg and an Emax = 65+/-0.2 ng GH/ml plasma. Again, this was very similar to GHRP-6 (ED50 = 3.9+/-1.4 nmol/kg and Emax = 74+/-7ng GH/ml plasma). GHRP-2 displayed higher potency but lower efficacy (ED50 = 0.6 nmol/kg and Emax = 56+/-6 ng GH/ml plasma). The specificity for GH release was studied in swine. None of the GH secretagogues tested affected FSH, LH, PRL or TSH plasma levels. Administration of both GHRP-6 and GHRP-2 resulted in increased plasma levels of ACTH and cortisol. Very surprisingly, ipamorelin did not release ACTH or cortisol in levels significantly different from those observed following GHRH stimulation. This lack of effect on ACTH and cortisol plasma levels was evident even at doses more than 200-fold higher than the ED50 for GH release. In conclusion, ipamorelin is the first GHRP-receptor agonist with a selectivity for GH release similar to that displayed by GHRH. The specificity of ipamorelin makes this compound a very interesting candidate for future clinical development. PMID- 9849824 TI - Methods for estimating HIV prevalence: A comparison of extrapolation from surveys on infection rate and risk behaviour with back-calculation for the Netherlands. AB - OBJECTIVES: To compare HIV prevalence estimates (total number infected) by using extrapolation from surveys on infection rate and risk behaviour (EIR) in specific segments of the population and back-calculation (BC) on reported AIDS cases. To discuss potential sources of bias and error, and to identify areas for improvement of the methodology. DESIGN: Systematic comparison and epidemiological assessment of data input, underlying assumptions, and output. METHODS: Low, possibly unbiased and high estimates of HIV prevalence as of January 1996 for homo/bisexual men, injecting drug users. heterosexual men and women with multiple partners, and blood transfusion recipients and haemophiliacs were derived from surveys and continuous data collections on HIV infection rate and risk behaviour in the Netherlands between 1992 and 1996. These were compared with estimates (point and 95 % CI) by empirical Bayesian BC on AIDS cases 1982-1995. RESULTS AND CONCLUSIONS: The estimate of HIV prevalence by EIR was 13,806 with low and high estimates of 9619 and 17,700, respectively. The HIV prevalence estimate by BC was 8812 (95% CI: 7759-9867). The available data from EIR are too limited for accurate estimates of HIV prevalence. EIR estimates could be improved considerably with more precise data on prevalence of risk behaviours and HIV prevalence rate for homosexual men. More confidence can be put in the BC estimates, but these could be underestimates because of the age effect on incubation time, pre-AIDS treatment and relapse of risk behaviour. BC estimates could be improved by a better representation of the incubation time distribution (including the effect of age there-upon), better data on the effectiveness and uptake of pre-AIDS antiretroviral treatment and prophylaxis of opportunistic infections, and on the level of underreporting. PMID- 9849825 TI - Changes in coronary risk profiles in employees after three years of multifactorial intervention. AB - This study was performed with the objective of assessing the effectiveness of the intervention at the work-site on lifestyle-related cardiovascular risk factors in four work-sites in Granada, Spain. Individualized face-to-face counseling was given to 1555 employees (1193 men and 362 women). After 3 years, a blinded assessment of the adequacy of the interventions was carried out. When changes in all employees were analysed together, ('per protocol' analysis) a non-significant reduction (-0.02%; 95% CI: -0.06 to +0.03%) in mean coronary risk was observed. We also distinguished between adequate and inadequate implementation of the intervention protocol. Observed changes in mean coronary risk were compared between both groups (adequate/inadequate implementation). Changes in mean coronary risk were beneficial only in the group that achieved an adequate level of intervention, with an observed significant mean reduction of risk of -0.12% (95% CI: -0.07 to -0.17) whereas changes were in the opposite direction in the other group with a mean change of +0.08% (95% CI: + 0.01 to + 0.16%). Work-site programs of health behaviour modification when appropriately implemented can attain although small, significant changes in cardiovascular risk profiles and thus be especially effective in achieving the population strategy of preventive cardiology. PMID- 9849826 TI - Associations between anthropometric indices of adiposity and atherogenic risk factors in Japanese working women aged 21-40 years. AB - This study was conducted to explore whether anthropometric indices of obesity are associated with atherogenic risk factors in young adult working women in Japan. The subjects were 492 women in an occupational setting. Predictor variables were body mass index (BMI), the sum of triceps and subscapular skinfold thickness (SFT), and the waist to hip ratio (WHR). Outcome variables were serum total cholesterol, triglyceride and blood pressures. The average age of the subjects was 26.3 (SD 3.9) years. The upper quartiles of BMI and SFT were significantly associated with all atherogenic risk factors, while the upper quartiles of WHR were not. Multiple comparisons revealed the 4th quartiles of BMI (> 22.25) and SFT (> 39 mm) to have significantly higher values for all atherogenic risk factors. We found that BMI and skinfold thickness were more relevant to the prediction of atherogenic risk factors than WHR in young adult Japanese women. PMID- 9849827 TI - Habitual coffee consumption and blood pressure: A study of self-defense officials in Japan. AB - The study aims to examine the relationship between habitual coffee consumption and blood pressure. The subjects were 3336 male self-defense officials aged 48-56 years, who received a preretirement health examination at the Self-Defense Forces Fukuoka Hospital between October 1986 and December 1992. Average coffee intake in the past year was ascertained by a self-administered questionnaire. A significant inverse relation between habitual coffee consumption and blood pressure was found with and without adjustment for alcohol use, cigarette smoking, body mass index, glucose tolerance, and green tea intake. Green tea, another major source of caffeine intake in Japanese, was unrelated to blood pressure. The adjusted mean differences per cup of coffee consumed per day were -0.6 mmHg (95% confident interval [CI]: -0.9 to -0.3, p = 0.0001) in systolic blood pressure and -0.4 mmHg (95% CI: -0.5 to -0.2, p = 0.0002) in diastolic blood pressure. Habitual coffee drinkers had lower blood pressure than non-drinkers at any levels of alcohol use, cigarette smoking, obesity, and glucose intolerance. Our findings consolidate the previous observation that habitual coffee consumption was associated with lower blood pressure. PMID- 9849828 TI - Cigarette smoking and risk of cancers of the colon and rectum: a case-control study from Italy. AB - The relationship between smoking habit and risk of colon and rectal cancers was considered in a case-control study conducted between 1991 and 1996 in six Italian centers. Cases were 1225 patients below age 75 with histologically confirmed cancer of the colon and 728 with cancer of the rectum. Controls were 4154 patients admitted to hospital for a wide spectrum of acute, non-neoplastic diseases. Compared to neversmokers, the odds ratios (OR) for current smokers of 25 or more cigarettes/day was 0.90 for patients with colon and 0.86 for those with rectal cancer and those for ex-smokers were 1.02 and 1.09 for colon and rectal cancer, respectively. No increase in risk was found with duration of the habit, the OR for 40 or more years being 0.79 for colon and 0.87 for rectal cancer. Furthermore, no relationship was apparent with time since starting (the OR for 40 or more years were 0.94 for colon and 1.05 for rectal cancer), or age at starting (the OR for < 18 years were 1.02 for colon and 1.01 for rectal cancer), or for pack-years smoked (the OR for 40 or more pack-years were 0.93 for colon and 0.91 for rectal cancer) or time since stopping among ex-smokers. No increase in risk was found in smokers of > or = 15 cigarettes/day for 40 years or longer (OR: 0.93). No significant heterogeneity was found across strata of age at diagnosis, sex, education, physical activity at work, intake of alcohol, coffee, vegetables, total energy, and number of meals/day. Likewise, no significant association was apparent for various intestinal subsites. Thus, this study did not find cigarette smokers at higher risk for cancer of the bowel even after a long duration and a long period since starting. PMID- 9849829 TI - Diphtheria antitoxin levels in the 14-30-year age group in Italy. AB - In order to verify diphtheria immunity a seroepidemiological survey was performed in 1996-1997. Serum samples were obtained from 501 subjects 14 years old, recruited at 8 schools in Rome, and from 490 subjects 20-30 years old recruited from 15 Italian regions. Serum diphtheria antitoxin was titrated using the Vero cell assay. The minimum protection level of antitoxin was set at 0.01 IU ml-1. The results show that the younger population have a good immunity to diphtheria while a large proportion of young adults is devoid of protective levels of diphtheria antitoxin. Out of the 501 subjects 14 years old, 495 (98.8%) had a diphtheria antitoxin titre > or = 0.01 IU ml(-1). Only 6 (1.2%) teenagers were susceptible. Out of the 490 subjects 20-30 years old, 109 (22.2%) were susceptible, 381 (77.8%) had a diphtheria antitoxin titre > or = 0.01 IU ml(-1). The data stress for booster immunization at the end of junior high school. PMID- 9849830 TI - Identification of allergens in a selected group of asthmatics in Lebanon. AB - Anti-allergen antibodies were searched for by an Enzyme Immunoassay in the sera obtained from 60 patients with a clinical diagnosis of asthma. Allergic rhinitis was also present in 28 patients. The diagnosis of asthma was based on clinical criteria that include history and clinical evidence of airflow obstruction. Ten potential allergens (Bermuda grass, Olive tree, Parietaria, Alternaria, Cat hair dander, Dog hair dander, Mite DPT, Mugwort, Birch tree and Timothy grass) common to the Mediterranean area, were utilized. Twenty-five of 60 specimens were seropositive. Mite DPT was the allergen identified in 16 of the 25 seropositive specimens. Six of the 25 seropositive specimens reacted with more than one allergen. This may be due to the existence of similar antigenic determinant groups in the allergens used. There was no correlation between the occurrence of rhinitis in addition to asthma on one hand and the identified allergen on the other. Sixteen of the 25 seropositive patients were female. This observation is believed to be related to their occupation. Dog hair dander was identified as the allergen in 1 of the 25 seropositive patients. The low figure obtained was expected because keeping house pets is not a common practice in Lebanon. The 35 seronegative patients may belong to the intrinsic asthma group or the causative allergen in each case was not one of the 10 allergens used in this study. These results indicate that the house dust mite appears to be the most common allergen in the Lebanese asthmatic group studied. PMID- 9849831 TI - The prevalence of type 2 diabetes and gestational diabetes mellitus in an inner city multi-ethnic population. AB - Zeeburg', a multiethnic town borough in the Amsterdam-East region, has one of the city's highest rates of immigrants. In the total population of 19,825 Surinam (mainly Creole), Turkish, Moroccan, and Dutch adults the prevalence of known type 2 diabetes in 1994 and of gestational diabetes mellitus (GDM) between January 1992 and January 1997 was investigated. Based on World Health Organization (WHO) criteria of 1985, the age-standardized prevalence of type 2 diabetes was similar in men (6.4%; 95% confidence interval [CI]: 5.6-7.2) and women (6.4%: 95% CI: 5.8 7.0) for all ethnic groups combined. However, the age- and sex-standardized prevalence of type 2 diabetes was significantly greater in the non-Dutch inhabitants than in the Dutch inhabitants (17.3% [95% CI: 12.9-21.6] in Surinam inhabitants, 10.9% [95% CI: 9.7-12.2] in Turkish inhabitants, 12.4% [95% CI: 9.7 15.0] in Moroccan inhabitants, and 3.6% [95% CI: 3.2-3.9] in Dutch inhabitants). The odds ratios for type 2 diabetes for the separate immigrant groups relative to the Dutch group were 5.88 (95% CI: 4.54-7.69) for Surinam inhabitants, 4.00 (95% CI: 2.86-5.55) for Turkish inhabitants, and 4.17 (95% CI: 3.03-5.55) for Moroccan inhabitants. GDM was present in 2.59% of women of non-Dutch origin compared with 0.62% of women of Dutch origin. A significant positive association was found between the non-Dutch origin and the occurrence of GDM (chi2 = 6.7; p < 0.01). The study highlights a high prevalence of known type 2 diabetes and GDM in the immigrant inhabitants and emphasizes that appropriate interventions are necessarily with implications for health targets and capitation based budgets. PMID- 9849832 TI - Serotype and phage type distribution of salmonellas from human and non-human sources in Italy in the period 1973-1995. AB - According to the data collected at the Rome-based National Reference Centre for Enterobacteria, 266,547 Salmonella strains isolated from human sources (226,513) and from non human sources (40,034) were characterised in Italy during the period 1973-1995. The serotype of all isolates, and the antibiotype and phage type of selected isolates were determined. Human-source isolations grew from 4372 in 1973 to 12,310 in 1995: non-human source isolations, from 339 in 1973 to 3459 in 1995. Salm. Typhimurium ranked first in the list of the most common serotypes isolated from both sources in the period 1973-1988 except in the years 1975 and 1976 when it was overtaken by Salm. Wien. Since 1983 Salm. Enteritidis has been among the top ten isolates from animals, and ranked first in the list of isolates from humans in 1988 and from non human sources in 1991. During the last years the number of multidrug-resistant strains, mostly belonging to phage types 104 and 193 of Salm. Typhimurium has been rising. Salmonella strains have also been isolated from numerous extraintestinal infections, almost exclusively caused by Salm. Enteritidis and Typhimurium. PMID- 9849833 TI - Epidemiological explanation of an outbreak of gastro-enteritis in Sweden in the absence of detailed microbiological information. AB - Waterborne gastroenteritis outbreaks have often gone undetected or been incompletely defined in terms of source and extent. Methods which allow detection or clarification of such events are therefore useful. We describe the methods used to detect and investigate such an outbreak. In autumn 1996 high school absence rates and the rate of parents absent from work to care for sick children suggested a health problem in a Swedish town which had a history of unexplained outbreaks of gastrointestinal disease. A systematic sample of 300 households was surveyed by post. Respondents represented 10% of the total population of the town. Questions concerning symptoms and exposures were included. The same questionnaire was used in a nearby town as a control. Sixty four percent of respondents reported an acute gastrointestinal illness during a two month period. Diarrhoea (90%) and abdominal pain (88%) were the most frequent symptoms among the sick. Two percent of those sick sought medical care. Exposures associated with disease were being a member of a large household, young age, and consumption of water from the community water supply. Attack rate showed a dose response relationship with increasing frequency of water consumption. The peak incidence of gastrointestinal illness occurred shortly after raw water quality control data had shown a rise in indicator bacteria. Further analysis, dividing those infected into groups according to when they became ill and whether they were the first member of their household to fall ill, supported the hypothesis of primary cases being infected from the water supply with some secondary person to person spread. PMID- 9849835 TI - Most common clinical presentation of Q fever in a province in the north of Spain. PMID- 9849836 TI - Can the use of cesarean section be regulated? A review of methods and results. PMID- 9849834 TI - Prevention of nosocomial infection in a pediatric intensive care unit (PICU) through the use of selective digestive decontamination. AB - OBJECTIVE: To assess the effectiveness of selective digestive decontamination (SDD) on the control of nosocomial infection (NI) in critically ill pediatric patients. DESIGN: A prospective, randomized, non-blinded and controlled clinical microbiology study. SETTING: The pediatric intensive care unit (PICU) of a tertiary level pediatric university hospital. CRITERIA FOR INCLUSION: Patients 1 month to 14 years old, who underwent some kind of manipulation or instrumentation (mechanical ventilation, vascular cannulation, monitoring of intracranial pressure, thoracic or abdominal drainage, bladder catheterization, peritoneal dialysis, etc.) and/or presented a neurological coma requiring a stay in the PICU of 3 or more days. PATIENTS: Over a period of 2 years, 244 patients met the inclusion criteria; 18 patients were withdrawn because of protocol violation. The treatment group comprised 116 patients and the control group, 110 patients. INTERVENTION: The treatment group received a triple therapy of colimycin, tobramycin and nystatin administered orally or via nasogastric tube every 6 hours. All patients with mechanical ventilation or immune-depression received decontamination treatment of the oropharyngeal cavity with hexitidine (Oraldine 0.5 mg/ml) every 6-8 hours in accordance with the PICU's conventional protocol. METHOD: Up to 10 types of nosocomial infection were diagnosed following criteria of the Centers for Disease Control (CDC). The severity and manipulation of the patients on admission was assessed using the therapeutic intervention scoring system (TISS) and multi-organ system failure scores (MOSF). MEASUREMENTS AND MAIN RESULTS: UNIVARIANT ANALYSIS: SDD did not significantly reduce the incidence of NI, antibiotic use, the length of stay, or mortality; although a small percentage of respiratory and urinary tract infections was detected, catheter-related bacteremia was the most common infection. MULTIVARIANT ANALYSIS: Controlling the risk factors for each child through log regression showed that SDD acted as a protective factor for more than 90% of the sample with respect to the appearance of respiratory and urinary tract infections, reducing the risk of such infections to 1/5 and 1/3, respectively. CONCLUSIONS: SDD was effective in controlling respiratory and urinary tract infections in children admitted to the PICU, but it did not reduce the incidence of other types of nosocomial infection. PMID- 9849837 TI - Effect of endothelin-1 on spontaneous contractions and effects of nimodipine and isradipine on endothelin-1-induced contractions in myometrial strips isolated from normal pregnant and preeclamptic women. AB - BACKGROUND: To examine the effect of endothelin-1 (ET-1) on spontaneous contractile activity and the effects of nimodipine and isradipine on ET-1-induced contractile responses in myometrial strips isolated from normal pregnant and preeclamptic women. MATERIAL AND METHODS: . Isolated myometrial strips were obtained from seven normal pregnant and seven preeclamptic women undergoing elective cesarean section and the strips were mounted in organ baths for recording of isometric tension. The effect of increasing concentration of ET-1 on spontaneous contractions and effects of increasing concentration of nimodipine and isradipine on ET-1-induced contractions were recorded. RESULTS: ET-1 dose dependently (10(-11)-10(-8) M) increased the amplitude and frequency of spontaneous contractions in all myometrial strips obtained from normal pregnant and preeclamptic women. The increase in the amplitude of contractions in preeclamptic strips was significantly higher than that of normal strips. The increase in amplitude of contractions in normal and preeclamptic strips reached statistical significance beginning from the concentrations of 10(-9) M and 10( 11) M, respectively. ET-1 (10(-8) M) also increased the basal tone of all myometrial strips isolated from normal pregnant and preeclamptic women. When ET-1 (10(-8) M)-contracted myoinetrial strips were exposed to increasing concentrations of nimodipine (10(-6)-3x10(-5)M) and isradipine (10(-5)-3x10(-4) M), nimodipine and isradipine dose-dependently decreased the amplitude and frequency of contractions. CONCLUSIONS: The increase in contractile response with ET-1 was significantly higher in myometrial strips isolated from preeclamptic women compared to those of myometrial strips isolated from normal pregnant women. These increases in contractile response are at least in part mediated by dihydropyridine-sensitive calcium channels, since they were significantly reduced in the presence of increasing concentrations of nimodipine and isradipine. PMID- 9849838 TI - Effects of maternal prone position on the umbilical arterial flow. AB - OBJECTIVE: The objective of our study was to clarify the effect of the maternal prone position on the feto-placental flow. In this position, the uterine compression of the large maternal vessels was expected to be completely eliminated. METHODS: Twenty-three normal pregnant women were cross-sectionally examined in the supine, the left lateral, the right lateral and the prone position. The systolic/diastolic ratios for the umbilical artery were calculated for each position. RESULTS: The umbilical arterial systolic/diastolic ratio in the prone position significantly decreased compared with that in the supine position. The left and right lateral position showed no significant change in the systolic/diastolic ratio. CONCLUSION: The maternal prone position can provide complete relief of uterine compression of the large maternal vessels. PMID- 9849839 TI - Maternal vitamin-D deficiency in Pakistan. AB - OBJECTIVE OF THE STUDY: This study was performed to assess the vitamin D status of healthy Pakistani nursing mothers and their breastfed infants. METHODS: Seventy-one nursing mothers and their breastfed infants belonging to upper and lower socio-economic class were examined 6 weeks to 11 months after delivery. Serum 25-hydroxy vitamin D [25(OH)D], serum calcium, phosphorus and alkaline phosphatase were measured. RESULTS: The mean serum 25(OH)D in mothers was 36.7+/ 32.4 nmol/L and 41.25+/-35.4 nmol/ L in infants. Thirty-four (48%) mothers and 37 (52%) infants had levels less than 25 nmol/ L. Significantly higher levels were found in uneducated mothers (p=0.01), mothers of lower socio-economic class (p<0.001) and in those living in mud houses (p<0.001). A significant correlation was found between serum 25(OH)D levels of infants under three months of age and their mothers (p<0.01). CONCLUSIONS: High prevalence of vitamin D deficiency was found in nursing mothers and their infants predominantly in the upper socioeconomic class. PMID- 9849840 TI - Frequency of anal sphincter rupture at delivery in Sweden and Finland--result of difference in manual help to the baby's head. AB - BACKGROUND: Anal sphincter rupture is a serious complication of vaginal delivery and almost half the affected women have persistent defecatory symptoms despite adequate primary repair. During the past decade, the incidence of anal sphincter ruptures has been increasing in Sweden and is currently estimated to occur in 2.5% of vaginal deliveries. The aim of the study was to report the frequency of anal sphincter ruptures in two university hospitals in two Scandinavian countries, Malmo in Sweden and Turku in Finland, and analyze the potential determinants. METHODS: Retrospective analysis of a population of 30,933 deliveries (26,541 vaginal) during the years 1990 to 1994. RESULTS: The incidence of anal sphincter ruptures in Malmo, Sweden was 2.69%, and in Turku, Finland 0.36%. There were no significant population differences for the known risk factors (fetal weight, nulliparity or fetal head circumference). However, there is a difference in manual support given to the perineum and to the baby's head when crowning through the vaginal introitus between Malmo and Turku. The proportion of operative vaginal deliveries and abnormal presentations was significantly higher in Turku reflected in the lower Apgar score at 5 minutes and longer duration of second phase of labor. When high risk deliveries (operative vaginal delivery, abnormal presentation and newborns over 4,000 g) were excluded, the risk for anal sphincter ruptures was estimated to be 13 times higher in Malmo than in Turku. CONCLUSIONS: The difference in the incidence of anal sphincter rupture between Malmo, Sweden and Turku, Finland may be due to the difference in manual control of the baby's head when crowning. PMID- 9849841 TI - Vaginal sonosalpingography in the evaluation of tubal patency. AB - BACKGROUND: During the past few years, sonosalpingography has been suggested as the first-line method to study tubal patency. This study was launched in order to study the applicability of this method at our institution. METHODS: Thirty-two patients suffering from primary or secondary infertility were evaluated for tubal patency with sonosalpingography using a pediatric Foley urinary catheter and a combination of air and saline solution as a contrast medium. The uterine tubes were evaluated separately and the results were compared to the findings at laparoscopy and chromotubation performed independently. Four patients conceived before their scheduled laparoscopy and were excluded from the study. In addition, the patency of three Fallopian tubes could not be adequately evaluated, leaving altogether 53 uterine tubes that were evaluated by both methods. RESULTS: The findings of both methods agreed in 47 out of 53 tubes (concordance, 88.7%). The sensitivity of sonosalpingography in diagnosing tubal patency was 90.2% and the specificity 83.3%. The positive predictive value for tubal patency by sonosalpingography was 94.9% and the negative predictive value 71.4%. Adverse events of sonosalpingography included moderate to severe abdominal pain in three patients, one vasovagal reaction, and one case of shoulder pain. No infectious complications were recorded. CONCLUSIONS: The results confirm that sonosalpingography utilizing air and saline as a contrast medium is a reliable, simple and well-tolerated method to assess tubal patency in an outpatient setting. In addition, the procedure can be performed without prophylactic antibiotics using a regular pediatric Foley urinary catheter instead of an expensive hysterosalpingography catheter. PMID- 9849843 TI - Women's knowledge of and attitude towards organized cervical smear screening. AB - OBJECTIVE: To describe women's barriers to cervical screening by asking about their experience with and knowledge of smear tests and by examining the women's contact and relationship with their GP. SETTING: Questionnaires sent to women in Aarhus County, Denmark. METHODS: A case control study in a cohort. The cohort, 133,500 women, represents women aged 23-60 years who were invited to the organized cervical screening program between 1.10.90 and 1.4.94. A case group, non-attenders (n=694), who had not had a smear within the previous 42 months, and a control group, attenders (n=1131), who had at least one smear test within the previous 42 months, were compared with each other. Predictor variables in the two groups were analyzed by univariate and multivariate (logistic regression) analysis. RESULTS: Non-attenders more often had insufficient contact with the GP (having a bad relationship, feeling of not receiving sufficient information) and associated a gynecological examination with more psychological unpleasantness. Most non-attenders (65.5%) and attenders (88.5%) intended to attend next time they were invited for cervical screening, and resistance to mass screening on principle did not seem to have great influence on the number of non-attenders. Both non-attenders and attenders had poor knowledge of the smear test. No significant difference was found concerning the characteristics of the general practice used by non-attenders and attenders. CONCLUSIONS: The main barriers to regular cervical screening are insufficient contact with the GP and psychological unpleasantness associated with the gynecological examination. Resistance to cervical screening on principle does not seem to have a great influence on attendance. PMID- 9849842 TI - Osmotically-induced release of vasopressin and oxytocin in non-pregnant women- influence of estrogen and progesterone. AB - BACKGROUND: Circulating vasopressin and oxytocin are influenced by ovarian steroid blood levels, but the effect of estrogen and progestogen treatment on induced release of the posterior pituitary hormones is not clear. METHODS: Eight postmenopausal women who had not been on hormonal replacement therapy for at least two months were included in the study. The women were treated for four weeks with transdermal administration of estradiol-17beta in a daily dose of 100 microg with the addition of 5 mg tablets of medoxyprogesterone twice daily for the last two weeks. A 25 minute intravenous infusion of hypertonic saline (0.06 mg/kg/min) was given before hormonal treatment, and after two and four weeks with serial plasma sampling for assay of vasopressin and oxytocin. RESULTS: The mean basal concentration of vasopressin, which was 0.83+/-0.13 (SE) pmol/L before hormonal treatment, increased to a statistically significant degree after estradiol alone to 1.18+/-0.11 pmol/L and decreased after combined estrogen/progestogen treatment to 0.31+/-0.02 pmol/L. Sodium concentration and osmolality increased in a similar way during all three infusions, but the resultant increase in vasopressin concentration was significantly smaller and slower after treatment with estradiol alone than in the first experiment without pretreatment. The areas under the concentration curve for the second and third infusion were significantly smaller than when no hormone treatment was given. The induced hyperosmolality also caused a rise in oxytocin levels, but no influence of ovarian hormone treatment was observed. CONCLUSIONS: Ovarian hormone administration influences vasopressin secretion, affecting both the basal levels in plasma and the responses to an increase in plasma osmolality. The influence of ovarian hormones on oxytocin secretion is minimal. PMID- 9849844 TI - Prolonged catheterization after vaginal prolapse surgery. AB - OBJECTIVE: To evaluate the domestic use of a transurethral Foley catheter after discharge from the hospital in patients with urinary retention after vaginal prolapse surgery. METHODS: We reviewed 27 patients who used a Foley catheter at home after prolapse surgery. All patients were interviewed by postal questionnaire regarding their experience of this procedure. RESULTS: Of 263 vaginal prolapse operations 27 (11%) needed a prolonged catheterization period at home. In all but two patients micturition was successfully restored after removal of the catheter at a median of 13 days after discharge. Thirty-seven percent of the patients had experienced the catheterization period as unpleasant. Seventy nine percent experienced no longterm bladder emptying problems, the others only mild and occasionally. CONCLUSION: Domestic use of a transurethral Foley catheter is a valid alternative to intermittent or suprapubic catheterization when normal micturition is not easily restored after vaginal prolapse surgery. It is, however, experienced as relatively unpleasant by more than one third of the patients. PMID- 9849845 TI - Cough-induced leak-point pressure--a valid measure for assessing treatment in women with stress incontinence. AB - BACKGROUND: Presently available urodynamic methods are of little use for assessing the severity of urinary stress incontinence or for evaluating of treatment, Cough-induced leak-point pressure may prove to be a more useful urodynamic method in these respects. A vaginal anti-incontinence device was used to validate this new urodynamic method. AIM: The primary aim was to determine the extent to which cough-induced leak-point pressure was affected by the vaginal device. Secondary aims were to study the short-term effects of the vaginal device on leakage and urinary flow, and to relate the change in cough-induced leak-point pressure to the change in leakage as expressed by a short-term pad test. METHODS: In a prospective study of 22 women with a history of stress incontinence, the effect of a new vaginal anti-incontinence device (Conveen Continence Guard) on cough-induced leak-point pressure, a short-term pad test and urinary flow was studied. RESULTS: Cough-induced leak-point pressure increased from 99.9 to 138.9 cm H2O, while leakage, measured by a short-term pad test, decreased from 22.7 to 3.3 g when using the device. Urinary flow was not significantly reduced by the device. CONCLUSIONS: Cough-induced leak-point pressure is a valid, quantitative, dynamic measure of urethral closure function which can be used to study the effects of treatment in patients with stress incontinence. The vaginal device is effective for treatment of stress incontinence in the short-term, without reducing urinary flow. PMID- 9849846 TI - Treatment of endometrial carcinoma with high-dose-rate brachytherapy alone in medically inoperable stage I patients. AB - PURPOSE: To review the results of treatment with high-dose-rate brachytherapy alone in 228 patients with stage I endometrial carcinoma who are unfit for surgery. METHODS: All patients received an exclusive radiation therapy by means of high-dose-rate Iridium 192 intracavitary brachytherapy without additional external beam radiation. RESULTS: At 5 years, the overall survival rate was 59.7% and disease specific survival 85.4% at 10 years 30.2% and 75.1%. In clinical stage Ia disease specific survival was 88.6% at 5 years and 82.7% at 10 years, in stage Ib 80.2% and 63.4%, respectively (p<0.02). Disease specific survival was not affected by tumor grade or age. The rates of local control are related to the size of the uterus but not to the tumor grading. Intrauterine recurrence occurred in 17.5% but extrauterine pelvic relapse in only 0.4% of patients. The calculated probability of severe complications was 4.6% at 5 years. CONCLUSION: HDR brachytherapy alone achieves excellent disease specific survival rates in patients with medically inoperable stage I endometrial carcinoma. PMID- 9849847 TI - The value of intraoperative consultation (frozen section) in the diagnosis of ovarian neoplasms. AB - BACKGROUND: Frozen Section is an important diagnostic tool to determine the nature of ovarian masses. However, like other diagnostic tools, frozen section also has some pitfalls. We aimed to discuss the source and the nature of inaccuracies associated with this procedure. METHODS: In this retrospective study 360 cases of ovarian masses examined by frozen section were re-evaluated. The sensitivity, specificity and predictive values of frozen section diagnosis of ovarian tumors were calculated. The reasons for the erroneous frozen-section diagnoses were discussed. RESULTS: Overall diagnostic agreement for ovarian lesions was 94.2%. Disagreements were found in nine cases (2.5%). Diagnosis was deferred to permanent sections in 12 cases (3.3%). The sensitivity for malignant tumors was 93.1% and specificity was 99.2%. The sensitivity for benign tumors was 99.2% and specificity was 92.1%. Most problematic cases were mucinous tumors, followed by tumors resembling fibrothecomas, in addition sections without viable tissue fragments or presence of extensive hemorrhage and necrosis also obscured the frozen diagnosis. Another factor was the lack of an effective communication between the surgeon and the pathologist. CONCLUSIONS: For an effective usage of this method not only the pathologist but also the surgeons must know the pitfalls of this method and also there must be good communication between the pathologist and the surgeon. Especially deferred cases should be minimized by good communication. In fact it's an intraoperative consultation method that enables the pathologist to gather all the preoperative, intraoperative findings and to be familiar with the further treatment plan of the patient. PMID- 9849848 TI - New retroperitoneal culdoplasty and colpopexy at the time of laparoscopic total abdominal hysterectomy (L-TAH). AB - BACKGROUND: After a laparoscopic total abdominal hysterectomy, the safety of the translaparoscopic, prophylactic, retroperitoneal pouch of a Douglas reconstruction and vaginal vault suspension was evaluated. METHODS: This operation was adapted from the laparotomy approach and has been investigated from July, 1992, to July, 1996. Modest modifications have been introduced to meet laparoscopic technology requirements. Observational clinical trials have been conducted on 112 women. The surgery was performed through laparoscopy by suturing and tying method, with no vaginal approach. RESULTS: This clinical trial showed positive results for the operation, with no complications observed in this group of patients. After the operation, there were no symptoms or signs of a dysfunctional vagina, or pelvic relaxation. CONCLUSION: This preventive operation is safe and it restores the pelvic topography, functional anatomy, and it prevents post-hysterectomy vaginal dysfunction disorders. PMID- 9849850 TI - Successful pregnancies following laser-induced interstitial thermotherapy (LITT) for treatment of large uterine leiomyomas by a minimally invasive method. PMID- 9849849 TI - Successful pregnancies after recurrent abortions in mild essential thrombocythemia. PMID- 9849851 TI - The danger of neovascularization in invasive gestational trophoblastic disease. PMID- 9849852 TI - Laparoscopic assisted colpoiesis according to Davydov. PMID- 9849853 TI - Differential diagnosis of glandular proliferations in the prostate. A conventional and immunohistochemical approach. AB - A variety of small acinar lesions of the prostate can mimic prostate cancer in punch biopsies and in transurethral resection material. The first part of this review deals with differential diagnostic problems of the central and transition zone, including atypical adenomatous hyperplasia of the prostate, atrophic processes, sclerosing adenosis, basal cell hyperplasia, and low-grade adenocarcinoma. The second part deals with differential diagnostic problems in the peripheral zone: prostatic intraepithelial neoplasia, postatrophic hyperplasia, Cowper's glands, seminal vesicles, and ductal and intraductal carcinoma. Finally, atypical and small acinar proliferations are described. Diagnostic perspectives are discussed. proliferations (ASAP) that cannot be integrated into any of the well-established diagnostic entities [1, 16, 22, 41]. The relevant glandular proliferations of the central, transitional and peripheral zones of the prostate are discussed here with reference to the related carcinomas. PMID- 9849854 TI - Comparison of CD44 expression in early colorectal carcinomas of the de novo and ex adenoma types. AB - Small colorectal carcinomas without morphological evidence of origin from an adenoma have been called "de novo" carcinomas. As changes in the expression of the adhesion molecule CD44 and its variants have been described along the adenoma carcinoma sequence in colorectal carcinoma, we compared patterns of CD44 expression in early de novo and ex-adenoma colorectal carcinomas by staining specimens from a group of early (pT1) colorectal carcinomas by immunohistochemistry for CD44 (standard and variant forms v3, v5, v6, v7, v7/8, v10). We evaluated carcinoma, adenoma (ex-adenoma cases), transitional mucosal areas and apparently nonneoplastic mucosa peripheral to the lesions (when present). A marked increase was seen in numbers and intensity of standard and variant forms of CD44 in carcinomatous areas compared with nonneoplastic mucosa in both groups, with no significant difference between the groups. However, adenoma areas of the ex-adenoma cases and the transitional mucosa of the de novo carcinomas had nearly identical staining patterns. Together with data from other molecular studies, this may be interpreted as evidence for an adenoma-type precursor lesion in so-called de novo colorectal carcinomas. PMID- 9849855 TI - Alterations in tumour suppressor gene p53 correlate with inhibition of thrombospondin-1 gene expression in colon cancer cells. AB - If activation of the p53 gene is involved in the progression or metastasis of colon cancer, it may affect the angiogenic phenotype in vivo. To verify this hypothesis, we studied the correlation between p53 accumulation and expression of thrombospondin-1 (TSP1) in colon cancer specimens. Levels of TSP1 gene expression were estimated by Northern blotting in 65 colon cancers. Accumulation of p53 and the distribution of TSP1 protein were evaluated immunohistochemically. Various levels of TSP1 gene expression were seen in colon cancers, while p53 accumulation was confirmed in 42 of the 65 colon cancers. The level of TSP1 gene expression demonstrated a significant inverse correlation with p53 accumulation in colon cancer. Colon cancer cells expressed TSP1 protein and p53 accumulation reciprocally in the same nests. These results suggest that alterations in the tumour suppressor gene p53 may inhibit TSP1 expression in colon cancer. PMID- 9849856 TI - Helicobacter pylori infection produces reversible glycosylation changes to gastric mucins. AB - The protective ability of gastric mucins may depend largely on their oligosaccharide chains. We evaluated the effects of H. pylori infection on the glycosylation of gastric mucins. Gastric biopsy specimens from 20 H. pylori infected patients before and after cure of the H. pylori infection and 8 normal uninfected volunteers were examined by immunostaining for simple mucin-type glycoproteins and blood-group-related antigens bearing type 1 chain backbone. The immunoreactivity in different gastric compartments was evaluated. Simple mucin type glycoproteins and blood-group-related antigens were expressed in surface mucous cells. Simple mucin-type glycoproteins showed antrum-predominant expression in normal volunteers and were found in significantly fewer surface mucous cells in infected patients than in normal volunteers; their expression was restored after eradication of H. pylori. Sialyl Lewis(a) and Lewis(b) were expressed in fewer surface mucous cells after than before eradication. The patterns of glycosylation of gastric mucins vary in different gastric compartments and are reversibly altered by H. pylori infection. These alterations may affect the protective functions of gastric mucins. PMID- 9849857 TI - The influence of p53 and associated factors on the outcome of patients with oral squamous cell carcinoma. AB - In several tumour entities the immunohistochemical detection of p53 has proved to be a predictive factor for the survival of the patients. In this study the effector waf1 and the regulator mdm2 responsible for the inactivation of p53 were also determined in 156 tissue samples of primary squamous cell carcinomas in the oral cavity and oropharynx, their lymph node metastases, and the epithelium outside the invasively growing tumour from 107 patients. In this latter epithelium there was a significant correlation between grade of dysplasia and staining for p53 (P<0.01). In the dysplastic epithelium a significant correlation between p53, waf1, and mdm2 was shown (P<0.05). Differences in the immunohistochemical staining between different blocks of the tumour tissue and also between primary tumours and their lymph node metastases were revealed in 11 44% of cases, but there was no correlation with other variables, such as formation of lymph node metastases. In contrast to the conventional tumour grading and staging, no influence of any of the variables determined on survival or recurrence-free survival could be detected. It seems that p53 and associated factors are important in the early stages of cancerogenesis but not in further tumour progression and metastatic spread. PMID- 9849858 TI - Characterization and chemosensitivity of two human malignant peripheral nerve sheath tumour cell lines derived from a patient with neurofibromatosis type 1. AB - Two new cell lines, designated NMS-2 and NMS-2PC, were established in vitro from a malignant peripheral nerve sheath tumour (MPNST) in the right thigh and a retroperitoneal lesion of a 30-year-old man with neurofibromatosis type 1 (NF1). The NMS-2 cell line was derived from the first tumour, and the NMS-2PC cell line from a retroperitoneal metastatic tumour detected 9 months later. Cultured NMS-2 cells showed epithelioid features, while NMS-2PC cells showed fibroblast-like features. However, both cell lines were strongly positive for S-100 protein. The transplanted NMS-2 and NMS-2PC tumours showed the same histological features typical of MPNST. Chromosomal analysis revealed that only the NMS-2 cells had a t (1;2) chromosomal translocation. Chemosensitivity tests demonstrated that NMS-2PC cells were far more sensitive than NMS-2 cells to Adriamycin and etoposide, which had been used clinically. All-trans-retinoic acid induced a morphological change in NMS-2PC cells so that they were no longer fibroblast-like, but epithelioid cells. We believe the epitheloid components in the MPNST were derived from typical spindle cells. PMID- 9849859 TI - The process of ultrastructural changes from nuclei to apoptotic body. AB - There have been many reports on the formation of apoptotic bodies, but little is known about the cellular pathological processes and the morphological changes involved. We induced apoptotic cell death by administering nivalenol (NIV), a trichothecene mycotoxin produced by Fusarium species, and investigated the ultrastructural process of formation of apoptotic bodies. The thymus was examined by electron microscopy 6, 12, and 18 h after administration. Apoptotic cell death was induced in the thymus of NIV-treated mice. The nuclei became invaginated and pinched off to give fragments, and crescent-shaped spaces (CSS) were found around the nuclear envelopes of these cells at quite an early stage. In some of these spaces, myelin figures were observed. We divided the process of formation into four stages and characterized each of them. These are easily recognized in morphological stages and are also useful for clarifying the apoptotic mechanism. PMID- 9849860 TI - Alterations in the distribution of plasma fibronectin and the ultrastructure of podocytes in the peripheral glomerular loops in nephrotic rats. AB - Glomerular distribution of rat plasma fibronectin was examined during the course of puromycin (PAN)- and daunomycin (DM)-induced nephrosis. In control animals, fibronectin was detected in the mesangial matrix and along the glomerular basement membrane (GBM), closely associated with the plasma membrane of glomerular cells. In peripheral loops, immunoprecipitates were preferentially distributed in the laminae rarae externa and interna. Fibronectin was densely precipitated in a glomerulosclerotic lesion induced by DM at 8 weeks after the injection. In peripheral loops, loss and reconstruction of epithelial foot processes occurred in PAN nephrosis but the change was accompanied by negligible perturbation of fibronectin distribution in the lamina rara externa. In contrast, a remarkable decrease of fibronectin was observed in DM nephrosis, unrelated to the presence or absence of foot processes. The decrease in immunoreactivity for fibronectin in the lamina rara externa seemed to have no association with podocyte attachment to or detachment from the GBM. Plasma fibronectin distributed in the lamina rara externa is not directly involved in the modification of podocyte configuration or podocyte attachment, although its spatial distribution may have some functional significance for preserving the ultrastructure of the GBM. PMID- 9849861 TI - Changes in VEGF expression and in the vasculature during the growth of early stage ethylnitrosourea-induced malignant astrocytomas in rats. AB - Vascular endothelial growth factor (VEGF), a potent angiogenic and vascular permeability factor, may be important as a mediator of brain tumour progression. However, it is still not clear whether VEGF plays a causative role in the early stage of glioma development. We investigated the relationship between VEGF protein expression (as assayed by immunohistochemistry) and different morphological parameters reflecting tumour progression (tumour diameter, vascular density and vascular diameter) in tumours at various stages. As a tumour model, ethylnitrosourea (ENU)-induced rat malignant astrocytoma was used. Tumours were classified by size and level of vascularity estimated by the von Willebrand factor (vWF) staining. Tumours less than 10 mm in diameter were designated early stage neoplastic lesions. All 34 early astroglial tumours were found to be VEGF positive. Increase in the VEGF immunopositive rate of tumour cells correlated significantly with increase in vascular density and vascular diameter. We suggest that VEGF induces angiogenesis and growth of microvessels, promoting growth of the early stage malignant astrocytoma. PMID- 9849862 TI - Evidence for in vivo mitosis by granule-containing mast cells from canine mastocytomas. AB - Mast cell accumulations are generally considered to arise almost exclusively from the recruitment of non-granulated, bone-marrow-derived, precursor cells, with the stem cell factor (SCF) reported to play a crucial role in the growth, development and maturation of granulated mast cells within specific tissue sites. In this study dog mastocytoma specimens have been examined by both immunohistochemical and ultrastructural techniques, to demonstrate that fully granulated mast cells are capable of mitotic activity. Observations showing the formation of mitotic spindles, chromosome separation and cytokinesis all support the concept that granulated mast cells are capable of proliferative activity. The ability of mature granulated mast cells to replicate provides an alternative process for local increases in mast cell numbers, at least in canine mast cell tumours. Such observations suggest the possibility that normal or neoplastic human mast cells, fully granulated, have the potential to proliferate in specific tissue sites. PMID- 9849863 TI - Distribution of intrahepatic mast cells in various hepatobiliary disorders. An immunohistochemical study. AB - There is evidence that mast cells are involved in a number of pathophysiological processes. The significance of mast cells in hepatic fibrosis was examined in 28 patients with histologically normal livers, 34 with acute liver diseases, 51 with chronic liver diseases, and 59 with cholestatic biliary diseases, using immunostaining of the mast cell-specific proteinase, tryptase. Mast cells that were positive for tryptase and for chymase were significantly increased in frequency in fibrotic portal tracts and fibrous septa, particularly in cholestatic/biliary diseases. Mast cells were also increased in frequency around the fibrotic septal and intrahepatic large bile ducts and peribiliary glands of biliary diseases. However, they were less common or even rare in the sclerotic bile ducts and in scarred portal or septal fibrosis. More than half of these more numerous mast cells were positive for histamine, and some were also positive for basic fibroblast growth factor. These two substances were detectable by immunoelectron microscopic in the cytoplasmic granules of mast cells. In contrast, mast cell numbers were not significantly increased in acute viral or drug-induced hepatitis, or in zones 2 and 3 of the hepatic acinus with respect to pericellular and perivenular fibrosis in chronic liver diseases. These findings suggest that mast cells increase in number in cholestatic/biliary diseases, and to a lesser degree in chronic liver diseases, and are involved in the active fibrous enlargement of portal tract and fibrous septa formation and also in the fibrosis of the intrahepatic bile ducts as they display fibrosis-promoting factors such as tryptase, fibroblast growth factor and histamine. PMID- 9849864 TI - Autoimmune enteropathy in infants. Pathological study of the disease in two familial cases. AB - In two brothers with autoimmune enteropathy there was total villous atrophy in the small intestine and marked lymphoid cell infiltration in the lamina propria of the entire digestive tract, discovered at autopsy in one of these patients. In addition, the pancreas showed diffuse interstitial infiltration by lymphocytes. The liver was enlarged, with extensive haematopoiesis and cholestasis. Similar lesions in the digestive tract were noticed in the second boy, but on immunosuppressive therapy his diarrhoea gradually disappeared. When he was 16 months of age, percutaneous biopsies showed moderately aggressive chronic hepatitis and a focal interstitial lymphoid infiltrate in the kidney. After 3 years of immunosuppressive therapy (prednisone, cyclosporin), the child ate well and total parenteral nutrition was discontinued. The intestinal lesions had regressed but fibrotic lesions of the liver persisted. PMID- 9849865 TI - Dramatic development of quadriplegia in a young man while engaged in heavy manual labour: a post-traumatic spinal cord injury? AB - A healthy 16-year-old male suddenly developed paraesthesiae in his hands during heavy manual labour. He was able to walk to the nearest doctor's practice himself. The symptoms worsened dramatically, and within a short period of time cardiac arrest occurred. After resuscitation and intensive care, quadriplegia due to a high cervical cord lesion was diagnosed. The patient died 70 days later without any changes in the neurological symptoms. Post mortem revealed severe focal ischaemic lesions in the cervical spinal cord in a mixed arterial/venous distribution, but no evidence of direct traumatic changes in the spinal cord, the spine or the soft tissues. All other possible causes than trauma were excluded. The clinical development of the symptoms suggest that this spinal cord lesion should be classified as a secondary traumatic spinal cord injury caused by a subluxation of the cervical spine. The pathogenesis of post-traumatic ischaemic damage to the spinal cord appears to be related to localized hypercoagulability resulting in the formation of microthrombi. Impaired microcirculation in a limited area and for a limited period of time may have caused the irregularly distributed ischaemic necrosis. PMID- 9849866 TI - A cooperative model for protonmotive heme-copper oxidases. The role of heme a in the proton pump of cytochrome c oxidase. AB - Oxido-reductions of metal centers in cytochrome c oxidase are linked to pK shifts of acidic groups in the enzyme (redox Bohr effects). The linkage at heme a results in proton uptake from the inner space upon reduction and proton release in the external space upon oxidation of the metal. The relationship of this process to the features of the proton pump in cytochrome c oxidase and its atomic structure revealed by X-ray crystallography to 2.8-2.3 A resolution is examined. A mechanism for the proton pump of cytochrome c oxidase, based on cooperative coupling at heme a, is proposed. PMID- 9849867 TI - Albumin's role in steroid hormone action and the origins of vertebrates: is albumin an essential protein? AB - Albumin, the major serum protein, binds a wide variety of lipophilic compounds including steroids, other lipophilic hormones and phytochemicals that bind to hormone receptors. Albumin has a low affinity for these lipophilic compounds. However, due to albumin's high concentration in serum, albumin is a major carrier of steroids and lipophilic hormones and regulator of their access to their receptors. Moreover, albumin functions as a sink for phytochemicals, which prevents their binding to hormone receptors and other cellular proteins, protecting animals from disruptive phytochemical-mediated endocrine effects. We propose that these properties of albumin were important in protochordates and vertebrates about 550 to 520 million years ago, just before and during the Cambrian. At that time, animal body sizes and exposure to phytochemicals in food were increasing, and animals in which albumin expression was high had a selective advantage in surviving and reproducing in the presence of toxic phytochemicals. This hypothesis that albumin has essential function(s) in mammalian endocrine physiology can be tested by comparing the effects of phytochemicals in Nagase rats that have 1/1000 the normal albumin concentration or in mice in which the albumin gene is knocked out with those in normal rats and mice. PMID- 9849869 TI - Hyperphosphorylation induces structural modification of tau-protein. AB - The effect of hyperphosphorylation on the structural properties and conformational stability of bovine tau-protein was studied by means of circular dichroism and fluorescence lifetime techniques. Normal protein contains unusual secondary structure elements: extended left-handed helices. The structure of this protein was assumed to be of a 'tadpole' type - a globular C-terminal part with a long and rigid tail included in the extended left-handed helix. Either a decrease or an increase of pH induced only minor changes of the normal tau-protein surface. Hyperphosphorylation affected the extended part of the protein molecule; the decrease of pH in this case induced considerable structural rearrangements, and the conformation of the C-terminal part of the protein molecule was transformed into a molten globule-like state. PMID- 9849868 TI - Conformational analysis of the endogenous mu-opioid agonist endomorphin-1 using NMR spectroscopy and molecular modeling. AB - Endomorphin-1 (Tyr-Pro-Trp-Phe-NH2) is a highly selective and potent agonist of the mu-opioid receptor. To identify structural attributes unique to this opioid peptide and potential sites of recognition, a conformational analysis has been performed using multidimensional NMR and molecular modeling techniques. The spectroscopic results, derived from experiments in both DMSO and water, indicate that endomorphin-1 exists in the cis- and trans-configuration with respect to the Pro-omega bond in approximately 25% and 75% populations, respectively. In DMSO, the cis-configuration adopts a compact sandwich conformation in which the Tyr and Trp aromatic rings pack against the proline ring, whereas the trans-configuration adopts an extended conformation. Although non-random structure was not observed in water, condensed phase molecular dynamics calculations indicate that trans isomers dominate the population in this higher dielectric medium. Structural comparison of the cis- and trans-configurations with morphine and selective mu peptide ligands PL-017 and D-TIPP, as well as the delta-selective peptide ligands TIPP (delta-antagonist, mu-agonist) and DPDPE were also performed and suggest the trans-isomer is likely the bioactive form. A hypothesis is proposed to explain mu and delta-selectivity based on the presence of spatially distinct selectivity pockets among these ligands. PMID- 9849870 TI - Cloning and functional characterisation of the mouse P2X7 receptor. AB - We have isolated a 1785-bp complementary DNA (cDNA) encoding the murine P2X7 receptor subunit from NTW8 mouse microglial cells. The encoded protein has 80%, and 85% homology to the human and rat P2X7 subunits, respectively. Functional properties of the heterologously expressed murine P2X7 homomeric receptor broadly resembled those of the P2X7 receptor in the native cell line. However, marked phenotypic differences were observed between the mouse receptor, and the other P2X7 receptor orthologues isolated with respect to agonist and antagonist potencies, and the kinetics of formation of the large aqueous pore. PMID- 9849871 TI - The structure and expression of a preprohormone of a neuropeptide, Hym-176 in Hydra magnipapillata. AB - Hym-176 (APFIFPGPKVamide) is a novel myoactive neuropeptide which was identified in systematic screening of signaling peptides in Hydra magnipapillata. By using PCR and library screening, we cloned and sequenced a full length cDNA which encoded a preprohormone of Hym-176. In the preprohormone, a typical signal sequence, one copy of Hym-176 precursor peptide and one copy of precursor sequence of another novel peptide, Hym-357 (KPAFLFKGYKPamide), were detected. In situ hybridization analysis revealed a strong signal in peduncle neurons. Signals were also detected, though weaker, in neurons in the gastric region and around the mouth. No signals were detected in the two extremities of the body, tentacles and basal disk. The expression pattern is correlated with the distribution of Hym 176 and its myoactive function. PMID- 9849872 TI - Natural agonist enhancing bis-His zinc-site in transmembrane segment V of the tachykinin NK3 receptor. AB - In the wild-type tachykinin NK3A receptor histidyl residues are present at two positions in TM-V, V:01 and V:05, at which Zn2+ functions as an antagonist in NK1 and kappa-opioid receptors with engineered metal-ion sites. Surprisingly, in the NK3A receptor Zn2+ instead increased the binding of the agonist 125I [MePhe7]neurokinin B to 150%. [MePhe7]neurokinin B bound to the NK3A receptor in a two-component mode of which Zn2+ eliminated the subnanomolar binding mode but induced a higher binding capacity of the nanomolar binding mode. Signal transduction was not induced by ZnCl2 but 10 microM ZnCl2 enhanced the effect of neurokinin B. Ala-substitution of HisV:01 eliminated the enhancing effect of Zn2+ on peptide binding. It is concluded that physiological concentrations of Zn2+ have a positive modulatory effect on the binding and function of neurokinin B on the NK3A receptor through a bis-His site in TM-V. PMID- 9849873 TI - Synthesis and structure-function study about tenecin 1, an antibacterial protein from larvae of Tenebrio molitor. AB - Tenecin 1, an inducible antibacterial protein secreted in the larvae of Tenebrio molitor, has a long N-terminal loop and common structural feature of insect defensin family corresponding to cysteine stabilized alpha/beta motif. To study the function of the N-terminal loop and disulfide bridges, N-terminal loop deleted tenecin 1, reduced tenecin 1 and tenecin 1 were chemically synthesized and their activities were measured. N-terminal loop deleted tenecin and reduced tenecin 1 did not show antibacterial activity. Circular dichroism (CD) spectroscopy data revealed that the alpha-helical content of tenecin 1 and the other proteins increased in the presence of 50% (v/v) trifluoroethanol (TFE) and the alpha-helical content of tenecin 1 was much higher than that of the other proteins in buffer with or without 50% (v/v) TFE. These results suggest that disulfide bridges are necessary for the activity structure and the N-terminal loop plays an important role in the increase of alpha-helix in the membrane mimetic environment and the activity. PMID- 9849875 TI - Genetic analyses of the in vivo function of LolA, a periplasmic chaperone involved in the outer membrane localization of Escherichia coli lipoproteins. AB - The major outer membrane lipoprotein (Lpp) of Escherichia coli is released from the cytoplasmic membrane into the periplasm as a complex with LolA, a periplasmic chaperone, prior to the localization in the outer membrane. To determine whether or not LolA is generally involved in the outer membrane localization of lipoproteins in vivo, the chromosomal lolA gene was manipulated so as to be controlled by the lac promoter-operator. Depletion of LolA caused a severe growth defect, and impaired the outer membrane localization of Lpp and Pal, another outer membrane lipoprotein. Although LolA depletion did not immediately arrest the growth of cells lacking Lpp, disruption of the chromosomal lolA gene was lethal to the lpp strain, indicating that LolA is generally required for the outer membrane localization of lipoproteins, and therefore essential irrespective of the presence or absence of Lpp. PMID- 9849874 TI - An innovative sandwich ELISA system based on an antibody cocktail for gluten analysis. AB - A cocktail sandwich ELISA based on the employ of two monoclonal antibodies (MAbs) as coating antibodies and a third MAb conjugated to horseradish peroxidase has been developed for the analysis of gluten in foods. Given that each MAb displays a wide specificity spectrum for wheat, barley, rye and oats prolamins, their combination for ELISA ensures a high crossreactivity with most of the potentially toxic gliadin, hordein, secalin and avenin protein family. One of the unprecedented features of the cocktail sandwich ELISA is that it permits for the first time analysis of barley hordeins in foods, which is unattainable using conventional or commercial ELISA kits. Besides, gliadins, hordeins and secalins are recognised to the same extent. The system provides a high detection sensitivity for gliadins, hordeins, secalins and avenins (1.5, 0.05, 0.15 and 12 ng/ml, respectively). The working linear range comprises 3-100 ng/ml with a gliadin detection limit of 1.5 ppm. This limit of detection is even better than that demanded in the latest Codex recommendation, 10 ppm. Cocktail ELISA data were contrasted with those of commercial ELISA kits and confirmed by mass spectrometry, a non-immunological technique which provides evidence for the occurrence of false positive results with the commercial kits. PMID- 9849876 TI - Transformed but not normal hepatocytes express UCP2. AB - Uncoupling protein 2 (UCP2) expression in liver is restricted to non-parenchymal cells. By means of differential display screening between normal rat liver and H4IIE hepatoma cells we have isolated a cDNA clone encompassing part of UCP2 cDNA. Northern blot analysis revealed that UCP2 is expressed in some hepatocarcinoma cell lines, while it is absent in adult hepatocytes. UCP2 mRNA in H4IIE cells was downregulated when cells were cultured for 36 h in 0.1% serum and its expression was restored upon addition of 10% serum or phorbol esters. Hypomethylation of UCP2 was observed in transformed UCP2 expressing cells. Our results indicate that UCP2 is expressed in some hepatocarcinoma cell lines and that serum components may participate in maintaining elevated UCP2 levels. PMID- 9849877 TI - Bacillus subtilis DnaG primase stabilises the bacteriophage SPP1 G40P helicase ssDNA complex. AB - Purified Bacillus subtilis DnaG primase (predicted molecular mass 68.8 kDa) behaves as a monomer in solution. We demonstrate that DnaG physically interacts with bacteriophage SPP1 hexameric helicase G40P (G40P6) in the absence of ATP. G40P6-ATP forms an unstable complex with ssDNA, and by itself carries out ATP driven translocation along a ssDNA template with low processivity. The presence of DnaG in the reaction mixture increased the helicase activity of G40P6 about 3 fold, but not the ATPase activity. The results presented here suggest that the DnaG protein stabilises the G40P6-ssDNA complexes. PMID- 9849878 TI - Distribution of the mammalian-wide interspersed repeats (MIRs) in the isochores of the human genome. AB - The distribution of MIRs (mammalian-wide interspersed repeats) was investigated in 164 human sequences (> or = 100 kb), which were assigned, according to their GC level, to isochore families L, H1, H2 and H3. MIR elements, whose total number in the genome was estimated to be about 3.3 x 10(5), were found to be unevenly distributed in human isochores. The majority of MIRs (55%) were found in the L isochore family. In contrast, MIR density was highest in H2, closely followed by H1, whereas densities in L and H3 were 2- and 3-fold lower than in H2, respectively. For this reason, the assessment of MIR distribution by inter-repeat PCR led to an overestimation of MIR numbers in H2 isochore and an underestimation in L isochores. PMID- 9849880 TI - Constitutive association of JAK1 and STAT5 in pro-B cells is dissolved by interleukin-4-induced tyrosine phosphorylation of both proteins. AB - The bipartite human interleukin-4 (IL-4) receptor was functionally expressed in murine pro-B cells and activated by human IL-4 to evoke intracellular signaling. Mutual association of signal transducing proteins within the receptor complex was then studied in dependence of ligand stimulation. Besides ligand-induced receptor heterodimerization and contacts of the two IL-4 receptor subunits alpha and gamma with Janus kinases JAK1 and JAK3 a prominent constitutive binding between JAK1 and signal transducer and activator of transcription STAT5 was detected. Since both these proteins become phosphorylated in response to IL-4 receptor stimulation, the influence of tyrosine phosphorylation on their mutual contact was analyzed. Association of JAK1 and STAT5 was found to occur exclusively between unphosphorylated proteins. PMID- 9849879 TI - Phospholipase C cDNAs from sponge and hydra: antiquity of genes involved in the inositol phospholipid signaling pathway. AB - To know whether or not the set of genes involved in the inositol phospholipid signaling pathway already existed in the early evolution of animals, we carried out cloning of cDNAs encoding phospholipase Cs (PLCs) from Ephydatia fluviatilis (freshwater sponge) and Hydra magnipapillata strain 105 (hydra). We isolated two PLC cDNAs, PLC-betaS and PLC-gammaS, from sponge and three cDNAs, PLC-betaH1, PLC betaH2, and PLC-deltaH, from hydra. From the domain organization and the divergence pattern in the PLC family tree, the sponge PLC-betaS and PLC-gammaS and the hydra PLC-deltaH are possibly homologous to the vertebrate PLC-beta, PLC gamma and PLC-delta subtypes, respectively. A detailed phylogenetic analysis suggests that the hydra PLC-betaH1 and PLC-betaH2 are homologs of the vertebrate PLC-beta1/2/3/Drosophila PLC21 and the vertebrate PLC-beta4/Drosophila norpA, respectively. A phylogenetic analysis of the PLC family and the protein kinase C (PKC) family, together with that of the G protein alpha subunit (Galpha) family, revealed that the origin of the set of genes G(alpha)q, PLC, PKC involved in the inositol phospholipid signaling pathway is very old, going back to dates before the parazoan-eumetazoan split, the earliest branching among extant animal phyla. PMID- 9849881 TI - A p38 MAP kinase inhibitor regulates stability of interleukin-1-induced cyclooxygenase-2 mRNA. AB - The mechanism by which p38 mitogen-activated protein kinase (MAPK) regulates the induction of cyclooxygenase (COX)-2 by interleukin-1 (IL-1) has been investigated in HeLa cells. SB 203580, an inhibitor of p38 MAPK, in the range 0.1-1 microM inhibited IL-1-stimulated PGE2 (but not arachidonic acid) release and this was associated with inhibition of induction of COX-2 protein and mRNA. IL-1 stimulated COX-2 transcription in HeLa cells about 2-fold as judged by both reporter gene and nuclear run-on assays. The inhibitor had no significant effect on this. However, in cells previously stimulated with IL-1 it caused rapid destabilisation of COX-2 mRNA independently of on-going transcription. The results suggest a novel function for p38 MAPK in the regulation of mRNA stability. PMID- 9849882 TI - Selective involvement of caspase-3 in ceramide induced apoptosis in AK-5 tumor cells. AB - Ceramide, a product of sphingomyelin metabolism, is a novel lipid second messenger that mediates diverse cellular functions. The present study demonstrates the activation of caspase-3/CPP-32beta, during apoptosis induced by cell permeable exogenous ceramides, in AK-5 tumor, a spontaneously regressing rat histiocytoma. The apoptotic events were suppressed by the caspase-3 specific tetrapeptide inhibitor DEVD-CHO but not by the caspase-1 inhibitor YVAD-CMK. In cells overexpressing Bcl-2, a significant decrease in cell death was observed after exogenous addition of ceramides. Furthermore the processing of caspase-3 to its active form upon apoptotic stimulus, and the subsequent cleavage of the substrate PARP, suggested a central role for caspase-3 in the ceramide mediated apoptosis in AK-5 tumor cells. PMID- 9849883 TI - Kinetic and spectroscopic characterization of an intermediate peroxynitrite complex in the nitrogen monoxide induced oxidation of oxyhemoglobin. AB - Stopped-flow spectroscopy studies of the nitrogen monoxide mediated oxidation of oxyhemoglobin in the pH range 5-10.5 show that an intermediate can be characterized at alkaline pH. The rate of decay of this species to methemoglobin increases significantly with decreasing pH such that it does not accumulate in quantities large enough to be observed under neutral or acidic conditions. Kinetic and spectroscopic properties of this intermediate support its assignment as a methemoglobin peroxynitrite complex. PMID- 9849884 TI - Expression of specific white adipose tissue genes in denervation-induced skeletal muscle fatty degeneration. AB - Denervation of skeletal muscle results in rapid atrophy with loss of contractile mass and/or progressive degeneration of muscle fibers which are replaced to a greater or lesser degree by connective and fatty tissues. In this study, we show that denervated rabbit muscles are transformed into a white adipose tissue, depending on their fiber types. This tissue does express LPL, G3PDH and particularly the ob gene, a white adipose tissue-specific marker, and does not express the brown adipose tissue molecular marker UCP1 mRNA. PMID- 9849885 TI - Influence of endogenous GM1 ganglioside on TrkB activity, in cultured neurons. AB - We verified the hypothesis that changes in the endogenous GM1 ganglioside density in the environment of TrkB, receptor of brain-derived neurotrophic factor, can affect receptor activity, and focused on rat cerebellar granule cells expressing both GM1 and TrkB. Changes of the amount of GM1 associated to immunoprecipitated TrkB and of receptor tyrosine phosphorylation were evaluated after treatment with phorbol-12-myristate-13-acetate (1 microM, 7 min), reported to affect the plasma membrane distribution of endogenous gangliosides in the same cells. After treatment, the amount of GM1 associated to receptor and TrkB phosphorylation decreased by about 40%. The amount of associated GM1 decreased by about 33% also after concomitant treatment with phorbol ester and brain-derived neurotrophic factor, but in this case the neurotrophin was unable to enhance receptor tyrosine phosphorylation. These results for the first time suggest that changes in the amount of endogenous GM1 in the environment of TrkB can modulate receptor activity, and offer new clues for a better understanding of physiological and pathological events of the nervous system. PMID- 9849886 TI - Tyrosine kinase activity is associated with CD44 in human neutrophils. AB - CD44 appears to be involved in signal transduction, however, the mechanism of this function is unclear. Protein kinase activity was detected in neutrophils associated with CD44. Most of the protein kinase activity associated with these antigens was tyrosine kinase activity. Src family kinases lyn and hck were found to account for much of the associated tyrosine kinase activity. The data suggest that associated tyrosine kinase activity may play a role in signal transduction from CD44 in neutrophils to regulate other cell functions. PMID- 9849887 TI - Cooperative cyclic interactions involved in metal binding to pairs of sites in EF hand proteins. AB - This study focuses on a closed net of electron-pair donor-acceptor interactions, present in the core of all metal-bound EF-hand pairs, that link both metal ions across a short two-stranded beta-sheet. A molecular model based on the above cycle of interactions was studied using semi-empirical molecular orbital quantum mechanical methods. The calculations indicate that the interactions in the model cycle are cooperative, that is, that the interaction energy of the cyclic structure is greater than that of the sum of isolated interactions between its components. The cooperativity in this cycle can be attributed to an increase in the stability of the interactions resulting from a mutual polarisation of the associated groups. The predicted polarisation of the amide groups in the cycle is in agreement with experimental NMR 15N deshielding observed for these amide groups upon metal binding. Experimental observations of strengthening of the beta sheet hydrogen bonds are also consistent with the model calculations. By this mechanism, the binding of the first metal ion would enhance the binding of the second metal ion, and thus, the intradomain cooperativity in cation binding of calmodulin and related EF-hand proteins can be ascribed, at least partly, to this short-range molecular mechanism. PMID- 9849888 TI - Pyrimidinoceptor potentiation by ATP in NG108-15 cells. AB - Regulation of inositol phospholipid hydrolysis by UTP and UDP in neuroblastoma x glioma hybrid cell line NG108-15 was potentiated in the presence of ATP. The effect of ATP was dose dependent and shifted the EC50 value for these uracil nucleotides up to three powers of magnitude, having no influence on the maximal value of the response. Adenine nucleotides (ADP, AMP, adenosine 5'-O-(3 thiotriphosphate) (ATPgammaS), beta,gamma-methyleneadenosine 5'-triphosphate (betagammaMeATP), 3'-O-(4-benzoyl)benzoyl ATP (BzATP) and 3'-deoxyadenosine 5'-O (1-thio)triphosphate (dATPalphaS)) as well as adenosine, had no influence on the pyrimidinoceptor response. The potentiation effect was abolished by excess of EDTA. The results were in agreement with the hypothesis of pyrimidinoceptor affinity regulation via extracellular phosphorylation of the receptor protein, initiated by ATP. This mechanism may have physiological implication for functioning of uracil nucleotides as endogenous signaling molecules. PMID- 9849889 TI - Selective activation of G-protein subtypes by vertebrate and invertebrate rhodopsins. AB - We have quantitatively investigated specificities in activating G-protein subtype by bovine and squid rhodopsins to examine whether or not the phototransduction cascade in each of the photoreceptor cells is determined by the colocalization of a large amount of G-protein subtype (Gt or Gq). In contrast to the efficient activation of respective Gt and Gq, bovine and squid rhodopsins scarcely activated G-protein counterparts. Exchange of alpha- and betagamma-subunits of Gt and Gq indicated the critical role of the alpha-subunit in specific binding to respective rhodopsins. Thus the specific recognition of G-protein subtype by each rhodopsin is a major mechanism in determining the phototransduction cascade. PMID- 9849890 TI - Activation of muscarinic K+ current by beta-adrenergic receptors in cultured atrial myocytes transfected with beta1 subunit of heterotrimeric G proteins. AB - Muscarinic K+ channels (IK(ACh)) in native atrial myocytes are activated by betagamma subunits of pertussis toxin (Ptx)-sensitive heterotrimeric G proteins coupled to different receptors. betagamma subunits of Ptx-insensitive Gs, coupled to beta-adrenergic receptors, do not activate native IK(ACh). In atrial myocytes from adult rats transfected with rat brain beta1 subunit IK(ACh) can be activated by stimulation of beta-adrenergic receptors using isoprenaline. This effect is insensitive to Ptx. These findings demonstrate for the first time promiscuous (Ptx-insensitive) coupling of Gsbetagamma to GIRK channels in their native environment. PMID- 9849891 TI - Phosphorylation site independent single R-domain mutations affect CFTR channel activity. AB - We investigated CFTR channel activity of mature R-domain mutants showing single alterations at sites other than the predicted phosphorylation sites. All mutations were found in cystic fibrosis (CF) patients (H620Q, E822K and E826K). The macroscopic CFTR chloride conductance induced by phosphorylation was significantly enhanced in Xenopus oocytes injected with mRNA of H620Q but reduced in the E822K and E826K mutants compared to wild type CFTR. The anion permeability sequence for all three mutants was the same as that of wild type CFTR. Cell attached single channel studies in COS cells revealed that both open channel probability and/or the number of functional channels were either higher (H620Q) or lower (E822K and E826K) than in wild type CFTR. Single channel conductances were unchanged in all mutants. Our results suggest that additional sites in the R domain other than phosphorylation sites influence gating of CFTR channels. PMID- 9849892 TI - ACTA, a fluorescent analogue of thapsigargin, is a potent inhibitor and a conformational probe of skeletal muscle Ca2+-ATPase. AB - Thapsigargin is a highly potent and selective inhibitor of sarco-endoplasmic reticulum (SERCA) family of Ca2+-ATPases and a useful tool in research concerning the function of intracellular Ca2+ stores. We describe here a novel fluorescent derivative (8-O-(4-aminocinnamoyl)-8-O-debutanoylthapsigargin, termed ACTA) of this compound, acting as a Ca2+-ATPase inhibitor with a potency approaching that of thapsigargin. Binding of ACTA to the skeletal muscle sarcoplasmic reticulum vesicles results in a strong fluorescence enhancement, approximately 66% of which depends on ACTA association with Ca2+-ATPase. This specific component of ACTA fluorescence is sensitive to the E1-E2 conformational equilibrium of the pump. The combined properties of high potency and binding-dependent fluorescence suggest ACTA to be a useful probe for a range of studies involving the SERCA class of ATPases. PMID- 9849893 TI - Crystal structure of rabbit muscle creatine kinase. AB - The crystal structure of rabbit muscle creatine kinase, solved at 2.35 A resolution by X-ray diffraction methods, clearly identified the active site with bound sulfates surrounded by a constellation of arginine residues. The putative binding site of creatine, which is occupied by a sulfate group in this analysis, has been tentatively identified. The dimeric interface of the enzyme is held together by a small number of hydrogen bonds. PMID- 9849894 TI - Human D3 dopamine receptor in the medulloblastoma TE671 cell line: cross-talk between D1 and D3 receptors. AB - In search of a cell line in which the D3 dopamine receptor is expressed endogenously, we found that the neuron-derived human medulloblastoma cell line TE671 expresses the human D3 (hD3) and D1 (hD1) receptor, but neither the D2 or D4 receptors. Exposure of TE671 cells to the D3 agonist 7-OH-DPAT (DPAT), or to the D1 agonist SKF-38393 (SKF) increased the expression of hD3 or hD1 mRNA, respectively. Moreover, whereas DPAT had no effect on hD1 mRNA levels, stimulating the cells with SKF caused an increase in both hD1 and hD3 transcript levels. These results suggest (i) that following ligand stimulation, hD3 and hD1 receptors are upregulated to enhance their own receptor expression, and (ii) that upregulation of hD1 receptor transcripts leads to a stimulation of the hD3 dopamine receptor transcripts. PMID- 9849895 TI - Longevity of lobsters is linked to ubiquitous telomerase expression. AB - Mammals have high growth rates in embryonic and juvenile phases and no growth in adult and senescent phases. We analyzed telomerase activity in a fundamentally different animal which grows indeterminately. Lobsters (Homarus americanus) grow throughout their life and the occurrence of senescence is slow. A modified TRAP assay was developed and the lobster telomeric repeat sequence TTAGG was determined. We detected telomerase activities which were dependent on RNA and protein components, required dGTP, dATP and dTTP, but not dCTP. Telomerase products with a five nucleotide periodicity were generated. High telomerase activities were detected in all lobster organs. We conclude that telomerase activation is a conserved mechanism for maintaining long-term cell proliferation capacity and preventing senescence, not only in cellular models or embryonic life stages but also in adult multicellular organisms. PMID- 9849896 TI - Priming of human neutrophil superoxide generation by tumour necrosis factor-alpha is signalled by enhanced phosphatidylinositol 3,4,5-trisphosphate but not inositol 1,4,5-trisphosphate accumulation. AB - In human neutrophils, significant agonist-stimulated superoxide anion (O2-) release is observed only after exposure to a priming agent such as TNFalpha. We have investigated the potential for TNFalpha to modulate N-formyl-Met-Leu-Phe (fMLP)-triggered Ins(1,4,5)P3 and PtdIns(3,4,5)P3 accumulation. TNFalpha pretreatment did not affect basal or stimulated Ins(1,4,5)P3 levels but greatly upregulated fMLP-stimulated PtdIns(3,4,5)P3 accumulation, in a manner that matched, both temporally and in magnitude, the increase in O2- generation implying a possible role for PtdIns(3,4,5)P3 in signalling primed O2- release. PMID- 9849898 TI - Functional heterodimeric amino acid transporters lacking cysteine residues involved in disulfide bond. AB - The protein mediating system L amino acid transport, AmAT-L, is a disulfide linked heterodimer of a permease-related light chain (AmAT-L-lc) and the type II glycoprotein 4F2hc/ CD98. The Schistosoma mansoni protein SPRM1 also heterodimerizes with h4F2hc, inducing amino acid transport with different specificity. In this study, we show that the disulfide bond is formed by heavy chain C109 with a Cys residue located in the second putative extracellular loop of the multi-transmembrane domain light chain (C164 and C137 for XAmAT-L-lc and SPRM1, respectively). The non-covalent interaction of Cys-mutant subunits is not sufficient to allow coimmunoprecipitation, but cell surface expression of the light chains is maintained to a large extent. The non-covalently linked transporters display the same transport characteristics as disulfide bound heterodimers, but the maximal transport rates are reduced by 30-80%. PMID- 9849897 TI - Endomorphins fully activate a cloned human mu opioid receptor. AB - Endomorphins were recently identified as endogenous ligands with high selectivity for mu opioid receptors. We have characterized the ability of endomorphins to bind to and functionally activate the cloned human mu opioid receptor. Both endomorphin-1 and endomorphin-2 exhibited binding selectivity for the mu opioid receptor over the delta and kappa opioid receptors. Both agonists inhibited forskolin-stimulated increase of cAMP in a dose-dependent fashion. When the mu opioid receptor was coexpressed in Xenopus oocytes with G protein-activated K+ channels, application of either endomorphin activated an inward K+ current. This activation was dose-dependent and blocked by naloxone. Both endomorphins acted as full agonists with efficacy similar to that of [D-Ala2,N-Me-Phe4,Gly ol5]enkephalin (DAMGO). These data indicate that endomorphins act as full agonists at the human mu opioid receptor, capable of stimulating the receptor to inhibit the cAMP/adenylyl cyclase pathway and activate G-protein-activated inwardly rectifying potassium (GIRK) channels. PMID- 9849899 TI - Developmental changes of gene expression in heme metabolic enzymes in rat placenta. AB - Transcription levels of the non-specific delta-aminolevulinate synthase (ALAS-N) and heme oxygenase-1 (HO-1) in the placenta at the terminal stage of pregnancy were comparable to those in the female adult liver and in the spleen, respectively. Immunohistochemical studies demonstrated that both enzymes were exclusively expressed in the trophoblast. During gestation, transcript of ALAS-N slightly increased, while HO-1 mRNA significantly decreased. Induced acute fetal hypoxia resulted in an increase in ALAS-N mRNA and in a decrease in HO-1 mRNA. These findings indicate that placental heme metabolism is influenced by the oxygen supply. PMID- 9849900 TI - Hypoxia induces apoptosis in human neuroblastoma SK-N-MC cells by caspase activation accompanying cytochrome c release from mitochondria. AB - We have attempted to elucidate the mechanism of apoptotic cell death induced by hypoxia (very low oxygen conditions) in neuronal cells. Human neuroblastoma SK-N MC cells under hypoxic conditions resulted in apoptosis in a time-dependent manner estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent chromatin dye. Pretreatment with Z-Asp-CH2-DCB, a caspase inhibitor, suppressed the DNA ladder in response to hypoxia in a concentration-dependent manner. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm the involvement of caspase-3 during apoptosis, Western blot analysis was performed using anti-caspase-3 antibody. The 20- and 17-kDa proteins, corresponding to the active products of caspase-3, were generated in hypoxia challenged lysates in which processing of the full length form of caspase-3 was evident. With a time course similar to this caspase-3 activation, hypoxic stress caused the cleavage of PARP, yielding an 85-kDa fragment typical of caspase activity. In addition, caspase-2 was also activated by hypoxia, and the stress elicited the release of cytochrome c into the cytosol during apoptosis. These results suggest that caspase activation and cytochrome c release play roles in hypoxia-induced neuronal apoptosis. PMID- 9849901 TI - The Arabidopsis homologue of an eIF3 complex subunit associates with the COP9 complex. AB - The Arabidopsis COP9 complex is a multi-subunit repressor of photomorphogenesis which is conserved among multicellular organisms. Approximately 12 proteins copurify with the COP9 complex. Seven of these proteins are orthologues of subunits of the recently published mammalian COP9 complex. Four of the proteins show amino acid similarity to various subunits of the COP9 complex, eIF3 complex and 19S cap of the proteasome. We have studied one of these proteins in order to determine if it is a component of the COP9 complex. Arabidopsis p105 is highly similar to the p110 subunit of the human elF3. The p105 gene is induced during photomorphogenesis, and RNA and protein analysis reveal different tissue accumulation patterns. p105 is found in a large protein complex. p105 interacts in yeast with both COP9 and FUS6, two known components of the COP9 complex. Our results indicate that p105 is not a component of the COP9 core complex, though it may interact with components of the complex. PMID- 9849902 TI - Osmotic changes and ethanol modify TFF gene expression in gastrointestinal cell lines. AB - The gastrointestinal tract is exposed to environmental insult as a result of food intake or in pathological conditions such as diarrhoea, and is therefore protected by the mucus layer. As part of it, trefoil peptides (TFFs) are able to modify the visco-elastic properties of the mucus, protect against experimental ulceration, and promote repair of the epithelia. We investigated, using transient reporter gene assays and RT-PCR in the gastric carcinoma cell line MKN45 and colon carcinoma cell lines LS174T and HT29, whether ethanol and osmotic changes can modify transcriptional activity of TFFs. In a mild hypotonic environment (200 mosmol/l) all three TFF genes were up-regulated by at least a factor of 2. In hypertonic medium (400 mosmol/ll), TFF1 and TFF3 were down-regulated, whereas TFF2 was up-regulated by elevated concentrations of sodium or chloride in MKN45. Raising the osmolality by ethanol resulted in an up-regulation of TFF3 in both colon cell lines but not in the gastric cell line. We conclude that alteration in TFF gene expression is a response of gut epithelia to deal with osmotic forces and ethanol. PMID- 9849903 TI - A conserved domain for glycogen binding in protein phosphatase-1 targeting subunits. AB - The skeletal muscle glycogen-binding subunit (GM) of protein phosphatase-1 (PP1) is the founding member of a family of proteins that tether the PP1 catalytic subunit (PP1C) to glycogen and promote the dephosphorylation of glycogen synthase. A hydrophobic sequence (called here the VFV motif) is conserved among GM, the liver subunit GL, and the widely expressed subunits, PTG, R5 and U5. This study analyzed the role of this VFV motif in binding to glycogen and PP1C. Glutathione S-transferase (GST) fusions with the N-terminal domain of GM (GST GM(1-240)) and with the full length R5 protein (GST-R5) both bound to glycogen in a co-sedimentation assay. In contrast, GST itself did not bind to glycogen. A single residue substitution in GST-GM(1-240), F155A, reduced glycogen binding by 40%. Double residue substitutions V150A/F155A and F155A/V159A resulted in greater reductions (60-70%) in glycogen binding, showing these hydrophobic residues influenced the protein-glycogen interaction. The wild type and V150A/ F155A fusion proteins were digested by trypsin into the same sized fragments at the same rate. Furthermore, the wild type and mutated GST-GM proteins as well as GST R5 bound equivalent amounts of PP1C, in either pull-down or far-Western assays. These results demonstrated retention of overall tertiary structure by the mutated fusion proteins, and indicated that glycogen and PP1C binding are independent of one another. A 68 residue segment of R5 encompassing the VFV motif was sufficient to produce glycogen binding when fused to GST. This motif, that is in bacterial and fungal starch metabolizing enzymes, probably has been conserved during evolution as a functional domain for binding glycogen and starch. PMID- 9849904 TI - Identification and subcellular localization of a novel Cu,Zn superoxide dismutase of Mycobacterium tuberculosis. AB - Periplasmic copper, zinc superoxide dismutases (Cu,ZnSOD) of several Gram negative pathogens have been shown to play an important role in protection against exogenous superoxide radicals and in determining virulence of the pathogens. Here we report the cloning and characterization of the sodC gene, encoding Cu,ZnSOD, from the Gram-positive Mycobacterium tuberculosis. The predicted protein sequence contains 240 amino acids with a putative signal peptide at the N-terminus and shows approximately 25% identity to other bacterial sodC. Recombinant proteins of a full-length sodC and a truncated form lacking the putative signal peptide were overexpressed in Escherichia coli and affinity purified. Renatured recombinant M. tuberculosis sodC protein possessed characteristics of a Cu,ZnSOD. Immunoblotting with an antiserum against the recombinant M. tuberculosis Cu,ZnSOD allowed detection of a single polypeptide in the lysate of M. tuberculosis. This polypeptide has a similar size as the recombinant protein without the putative signal peptide indicating that the endogenous Cu,ZnSOD in M. tuberculosis might be processed and secreted. Furthermore, immunogold electron microscopic image showed that Cu,ZnSOD is located in the periphery of M. tuberculosis. The enzymatic activity and subcellular localization of this novel Cu,ZnSOD suggest that it may play a role in determining virulence of M. tuberculosis. PMID- 9849905 TI - Dissection of multi-protein complexes using mass spectrometry: subunit interactions in transthyretin and retinol-binding protein complexes. AB - Complexes formed between transthyretin and retinol-binding protein prevent loss of retinol from the body through glomerular filtration. The interactions between these proteins have been examined by electrospray ionization combined with time of-flight mass analysis. Conditions were found whereby complexes of these proteins, containing from four to six protein molecules with up to two ligands, are preserved in the gas phase. Analysis of the mass spectra of these multimeric species gives the overall stoichiometry of the protein subunits and provides estimates for solution dissociation constants of 1.9 +/- 1.0 x 10(-7) M for the first and 3.5 +/- 1.0 x 10(-5) M for the second retinol-binding protein molecule bound to a transthyretin tetramer. Dissociation of these protein assemblies within the gas phase of the mass spectrometer shows that each retinol-binding protein molecule interacts with three transthyretin molecules. Mass spectral analysis illustrates not only a correlation with solution behavior and crystallographic data of a closely related protein complex but also exemplifies a general method for analysis of multi-protein assemblies. PMID- 9849906 TI - Detection of non-covalent interaction of single and double stranded DNA with peptides by MALDI-TOF. AB - DNA-histone interaction facilitates packaging of huge amounts of DNA in the confined space of the nucleus. The importance of this interaction underscores the need for new analytical techniques to acquire a better understanding of nuclear dynamics. Electrospray-ionization mass spectrometry made it possible to investigate non-covalently-bound biopolymers. We are enlarging the scope of available analytical tools by studying non-covalent interaction between single and double stranded DNA and peptides with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The interaction is an ionic one, between the negatively charged sugar-phosphate backbone of single stranded DNA and positively charged side chains of Arg- and Lys-rich peptides as demonstrated by Vertes' group with the dipeptides Arg-Lys and His-His. We replicated Lecchi and Pannell's work, which showed that double stranded DNA could be seen by MALDI using 6-aza-2-thiothymine (ATT) as matrix. We tried various peptides and found that as was demonstrated in DNA-histone interaction, a certain ratio and arrangement of basic residues was needed in order to generate ionic binding between DNA and peptide. We tested various single and double stranded DNA with the peptide of choice, and found that other variables such as pH value of solution, ionic strength, and matrix system did play a role. PMID- 9849907 TI - Stability of secondary structural elements in a solvent-free environment. II: the beta-pleated sheets. AB - The stability of single beta-strands and multistrand beta-pleated sheets as elements of secondary structure is examined in the absence of intermolecular interactions. Such experimental conditions (e.g., complete removal of solvent molecules and counterions) are achieved by placing the peptide ions in the gas phase. The metastable multiply- charged peptide ions produced by electrospray ionization undergo unimolecular dissociation. Intercharge repulsion within the precursor ions gives rise to the elevated kinetic energy of fragment ions, which is measured using Mass-analyzed Ion Kinetic Energy (MIKE) spectrometry. Intercharge distances calculated based on these measurements are compared to the numbers derived from molecular mechanics calculations with charge site assignments based on relative proton affinities. Evidence is presented suggesting that single beta-strands form collapsed structures in the absence of solvents, while multistrand beta-pleated sheets are likely to retain "native-like" secondary structures under the same conditions. These results indicate that intramolecular hydrogen bonds are the major factor determining the three dimensional arrangements of polypeptides in the gas phase, compensating both long and short-range electrostatic repulsions. This is in good agreement with our earlier findings (Proteins 27:165170, 1997) concerning stability of helical conformation of melittin in the absence of solvent. PMID- 9849908 TI - Application of electrospray ionization mass spectrometry for studying human immunodeficiency virus protein complexes. AB - Mass spectrometry (MS) with electrospray ionization (ESI) has shown utility for studying noncovalent protein complexes, as it offers advantages in sensitivity, speed, and mass accuracy. The stoichiometry of the binding partners can be easily deduced from the molecular weight measurement. In many examples of protein complexes, the gas phase-based measurement is consistent with the expected solution phase binding characteristics. This quality suggests the utility of ESI MS for investigating solution phase molecular interactions. Complexes composed of proteins from the human immunodeficiency virus (HIV) have been studied using ESI MS. Multiply charged protein dimers from HIV integrase catalytic core (F185K) and HIV protease have been observed. Furthermore, the ternary complex between HIV protease dimer and inhibitor pepstatin A was studied as a function of solution pH. Zinc binding to zinc finger-containing nucleocapsid protein (NCp7) and the NCp7-psi RNA 1:1 stoichiometry complex was also studied by ESI-MS. No protein-RNA complex was observed in the absence of zinc, consistent with the role of the zinc finger motifs for RNA binding. PMID- 9849909 TI - Conformational mapping of a viral fusion peptide in structure-promoting solvents using circular dichroism and electrospray mass spectrometry. AB - The N-terminal domain of human immunodeficiency virus (HIV)-1 glycoprotein 41,000 (FP; residues 1-23; NH2-AVGIGALFLGFLGAAGSTMGARS-CONH2) is involved in the fusion and cytolytic processes underlying viral-cell infection. Here, we use circular dichroism (CD) spectroscopy, along with electrospray ionization (ESI) mass spectrometry and tandem (MS/MS) mass spectrometry during the course of hydrogen/deuterium exchange, to probe the local conformations of this synthetic peptide in two membrane mimics. Since amino acids that participate in defined secondary structure (i.e., alpha-helix or beta-sheet) exchange amido hydrogens more slowly than residues in random structures, deuterium exchange was combined with CD spectroscopy to map conformations to specific residues. For FP suspended in the highly structure-promoting solvent hexafluoroisopropanol (HFIP), CD spectra indicated high alpha-helix and disordered structures, whereas ESI and MS/MS mass spectrometry indicated that residues 5-15 were alpha-helical and 16-23 were disordered. For FP suspended in the less structure-promoting solvent trifluoroethanol (TFE), CD spectra showed lower alpha-helix, with ESI and MS/MS mass spectrometry indicating that only residues 9-15 participated in the alpha helix. These results compare favorably with previous two-dimensional nuclear magnetic resonance studies on the same peptide. PMID- 9849910 TI - Characterization of the folding pathway of recombinant human macrophage-colony stimulating-factor beta (rhM-CSF beta) by bis-cysteinyl modification and mass spectrometry. AB - Melarsen oxide [p-(4,6-diamino-1,3,5-triazin-2-yl)aminophenylarsonous acid (MEL)], which selectively bridges spatially neighboring bis-cysteinyl residues in (reduced) proteins, was used to trap folding intermediates chemically during 1) time-dependent renaturation of recombinant human macrophage colony-stimulating factor (rhM-CSF); by redox refolding in vitro; 2) reductive unfolding in the presence of the trapping reagent; and 3) denaturing unfolding reactions in urea and guanidinium hydrochloride. Characterization of intermediates from folding and unfolding reactions was performed by electrospray ionization mass spectometry (ESI-MS). In all folding and unfolding reactions a characteristic dimeric intermediate with two attached melarsen oxide (MEL) groups was observed, suggesting that these rhM-CSF beta species were important refolding intermediates. These intermediates presented a characteristic "charge structure" in ESI spectra with a most abundant 26+ charged molecular ion whereas the mature homodimeric rhM-CSF beta showed a most abundant 23+ molecular ion, indicating that the final product was more compact. The major locations of the two MEL groups were identified by mass spectrometric peptide mapping at cysteine residues C157 and C159 from each monomer. Cysteine residues C7 and C90 were minor modification sites. The mass spectrometric results from the in vitro folding reactions of rhM-CSF beta are in agreement with intrinsic tryptophan fluorescence measurements and are consistent with the folding pathway that starts with a fully reduced monomer (R), includes partially folded monomeric intermediates (M) and dimeric intermediates (D), and yields a final product with the native tertiary structure (N): 2R ==> 2M ==> D ==> N. Our results show that selective chemical trapping of bis-thiol groups of proteins with MEL permits study of folding pathways by mass spectrometric structure characterization of intermediates with otherwise transient conformations. PMID- 9849911 TI - Mass spectrometric mapping of ion channel proteins (porins) and identification of their supramolecular membrane assembly. AB - Mass spectrometric peptide mapping, particularly by matrix-assisted laser desorption-ionization (MALDI-MS), has recently been shown to be an efficient tool for the primary structure characterization of proteins. In combination with in situ proteolytic digestion of proteins separated by one- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), mass spectrometric peptide mapping permits identification of proteins from complex mixtures such as cell lysates. In this study we have investigated several ion channel membrane proteins (porins) and their supramolecular assembly in mitochondrial membranes by peptide mapping in solution and upon digestion in the gel matrix. Porins are integral membrane proteins serving as nonspecific diffusion pores or as specific systems for the transport of substrates through bacterial and mitochondrial membranes. The well-characterized porin from Rhodobacter capsulatus (R.c.-porin) has been found to be a native trimeric complex by the crystal structure and was used as a model system in this study. R.c.-porin was characterized by MALDI-MS peptide mapping in solution, and by direct in situ-gel digestion of the trimer. Furthermore, in this study we demonstrate the direct identification of the noncovalent complex between a mitochondrial porin and the adenine nucleotide translocator from rat liver, by MALDI-MS determination of the specific peptides due to both protein sequences in the SDS-PAGE gel band. The combination of native gel electrophoresis and mass spectrometric peptide mapping of the specific gel bands should be developed as a powerful tool for the molecular identification of protein interactions. PMID- 9849912 TI - Mass spectrometric identification and microcharacterization of proteins from electrophoretic gels: strategies and applications. AB - The entire genomic DNA sequences of a number of prokaryotic and eukaryotic species are now available and many more, including the human genome, will be completed in the near future. The state-of-life of a cell at any given time, however, is defined by its protein composition, i.e., its proteome. Gel electrophoresis, mass spectrometry, and bioinformatics will be important tools for protein and proteome analysis in the post-genome era. Protein identification from electrophoretic gels by mass spectrometric peptide mapping or peptide sequencing combined with sequence database searching is established and has been applied to numerous biological systems. We describe current strategies and selected applications in molecular and cell biology. The next challenges are detailed structure/function analyses, which include studying the molecular composition of multiprotein complexes and characterization of secondary modifications of proteins. The advantages and limitations of a number of mass spectrometry-based strategies designed for microcharacterization of low amounts of protein from electrophoretic gels are discussed and illustrated by examples. PMID- 9849913 TI - Pulsed electromagnetic fields enhance the induction of cytokines by peripheral blood mononuclear cells challenged with phytohemagglutinin. AB - We evaluated the effects of a 50-Hz pulsed electromagnetic field on the production of cytokines by both resting and mitogen-treated peripheral blood mononuclear cells. Our results demonstrate that after exposure of normal cells to EMFs for 12 h, the levels of neither interleukin-1beta, nor interleukin-2 were increased. Indeed, the concentration of tumor necrosis factor alpha decreased significantly immediately after the exposure period. The results were, however, markedly different when cells were stimulated with phytohemagglutinin immediately before the exposure to EMFs. In this case the levels of cytokines, measured 24 and 48 h after the treatment, were 630 +/- 440 pg/ml and 910 +/- 530 pg/ml for interleukin-1beta, 530 +/- 330 pg/ml, and 860 +/- 560 pg/ml for tumor necrosis factor alpha, respectively. These values were significantly higher (P < 0.05) when compared with the controls. Interleukin-2 levels were significantly higher at the end of the EMF exposure only in supernatants of phytohemagglutinin stimulated cells and, as a consequence of this increase, the proliferation indexes also were significantly increased 48 h after the EMFs' treatment. The comparison between biological activity and the cytokine antigen present in our samples indicated that the amount of antigen was paralleled by an equal recovery of biological activity. This suggests either the absence of qualitative differences in these proteins or the impairment of both the transcriptional and translational processes. PMID- 9849914 TI - Response of cytosolic calcium, cyclic AMP, and cyclic GMP in dimethylsulfoxide differentiated HL-60 cells to modulated low frequency electric currents. AB - The action of interferential current (IFC), an amplitude-modulated 4000 kHz current used in therapeutic applications, upon intracellular calcium, adenosine 3':5'-cyclic monophosphate (cAMP), and guanosine 3':5'-cyclic monophosphate (cGMP) was investigated. Human promyelocytes (HL-60) were differentiated to granulocytes by dimethylsulfoxide (DMSO) treatment and exposed for 5 min at 25, 250, and 2500 microA/cm2 current density. No significant changes in cytosolic free calcium were detected as a function of modulation frequency of the IFC. However, intracellular cAMP reacted in a complex way to modulation frequency, resulting in stimulations and depressions within the range of frequencies studied (0-125 Hz). The "windows" of modulation frequency, where statistically significant increases or decreases in cAMP were noted, coincided with those published earlier for mouse fibroblasts. Cellular cGMP content was always lowered by IFC treatment. Furthermore, no significant influence of IFC current density upon the three second messengers was noted. These results, which also include data relating to treatment with sinusoidal 50 Hz current, contribute to a more detailed understanding of the primary biophysical mechanisms of signal transduction by time-varying electric fields. PMID- 9849915 TI - A compact shielded exposure system for the simultaneous long-term UHF irradiation of forty small mammals: I. Electromagnetic and environmental design. AB - To carry out in vivo studies of the possible health effects of radiation from cellular telephone handsets, it is necessary to expose large numbers of small mammals at realistic power densities, modulations, and frequencies. Because even microwatt leakage could compromise the local cellular system, extreme care in shielding is required. Experimental logistics dictate, however, that the irradiated animals be easily accessed and that it be possible to irradiate them in small groups, while other groups are being loaded into or unloaded from the irradiators. This problem has been resolved by exposing the animals in aluminum sheathed rectangular parallelepipeds, lined with microwave absorber and having doors that can be opened readily. Inside each of these microwave anechoic "chamberettes" is a vertical, four-element collinear array of dipole antennas; and around each antenna, 10 animal restrainers can be arranged like spokes on a wheel. The system has worked efficiently in studies of up to 480 rats. There is negligible coupling between antennas, and back reflection at an antenna's feed line is down 7-9 dB. Received CDMA power at the local base station is below the receiver's noise floor. Interior illumination reinforces the rats' diurnal rhythms, and the rats sleep during irradiation. Experimental logistics are excellent. In this paper, the irradiator design is presented. PMID- 9849916 TI - Protein kinase C activity following exposure to magnetic field and phorbol ester. AB - We examined the separate and combined effects of 60 Hz sinusoidal magnetic fields (MFs) and a phorbol ester on protein kinase C (PKC) activity in HL60 cells. No enhancement in PKC activity was observed when a cell culture was exposed to a 1.1 mT (rms) MF alone or to a combination of MF and 2 microM phorbol 12-myristate 13 acetate (PMA) for 1 h. In a second set of experiments, cells were preexposed to a less than optimal concentration of PMA (50 nM) for 45 min, followed by a 15 min exposure to both PMA and MF. The data showed a greater decrease in cytosolic PKC activity and a larger increase in membrane activity than was induced by either 1 h PMA treatment alone or PMA and sham MF exposure. One logical conclusion from these data is that MFs may be acting in a synergistic manner on a pathway that has already been activated. Therefore, we suggest that MFs, rather than producing biological effects by a new pathway or mechanism of interaction, exert their effect(s) by interacting with already functioning reactions or pathways. If correct, the question of an MF's mechanism of interaction refocuses on how weak fields might enhance or depress a molecular reaction in progress, rather than on finding a new transduction pathway. PMID- 9849917 TI - Effects of low-frequency magnetic fields on fetal development in CBA/Ca mice. AB - Effects of alternating magnetic fields (MFs) on the embryonic and fetal development in CBA/Ca mice were studied. Mated females were exposed continuously to a sinusoidal 50 Hz (13 microT or 0.13 mT root mean square) or a sawtooth 20 kHz (15 microT peak-to-peak) MF from day 0 to day 18 of pregnancy for 24 h/day until necropsied on day 18. Control animals were kept under the same conditions without the MF. MFs did not cause maternal toxicity. No adverse effects were seen in maternal hematology and the frequency of micronuclei in maternal bone marrow erythrocytes did not change. The MFs did not increase the number of resorptions or fetuses with major or minor malformations in any exposure group. The mean number of implantations and living fetuses per litter were similar in all groups. The corrected weight gain (weight gain without uterine content) of dams, pregnancy rates, incidences of resorptions and late fetal deaths, and fetal body weights were similar in all groups. There was, however, a statistically significant increase in the incidence of fetuses with at least three skeletal variations in all groups exposed to MFs. In conclusion, the 50 Hz or 20 kHz MFs did not increase incidences of malformations or resorptions in CBA/Ca mice, but increased skeletal variations consistently in all exposure groups. PMID- 9849918 TI - 50 Hz magnetic field effects on the performance of a spatial learning task by mice. AB - Intense magnetic fields have been shown to affect memory-related behaviours of rodents. A series of experiments was performed to investigate further the effects of a 50 Hz magnetic field on the foraging behaviour of adult, male C57BL/6J mice performing a spatial learning task in an eight-arm radial maze. Exposure to vertical, sinusoidal magnetic fields between 7.5 microT and 7.5 mT for 45 min immediately before daily testing sessions caused transient decreases in performance that depended on the applied flux density. Exposure above a threshold of between 7.5 and 75 microT significantly increased the number of errors the animals made and reduced the rate of acquisition of the task without any effect on overall accuracy. However, the imposition of a 45-minute delay between exposure at 0.75 mT and behavioural testing resulted in the elimination of any deficit. Similarly, exposure to fields between 7.5 microT and 0.75 mT for 45 min each day for 4 days after training had no amnesic effects on the retention and subsequent performance of the task. Overall, these results provide additional evidence that 50 Hz magnetic fields may cause subtle changes in the processing of spatial information in mice. Although these effects appear dependent on field strength, even at high flux densities the field-induced deficits tend to be transient and reversible. PMID- 9849919 TI - No effects of DC and 60-Hz AC magnetic fields on the first mitosis of two species of sea urchin embryos. AB - The early divisions of sea urchin eggs was used as a model to study the effects of static and of 60 Hz sinusoidal magnetic fields. Two species were used (Sphaerechinus granularis and Paracentrotus lividus). Eggs were fertilized and exposed in two separate coils to the fields (up to 8 mT). Great care was taken to control the temperature of each sample. No difference was found in the time of the first division that could not be attributed to a temperature difference between samples. Comparison is made with other published data on various species. PMID- 9849920 TI - A simple experiment to study electromagnetic field effects: protection induced by short-term exposures to 60 Hz magnetic fields. AB - Stress proteins are important in protection during cardiac ischemia/reperfusion (cessation and return of blood flow) and are reportedly induced by electromagnetic (EM) fields. This suggests a possible ischemia protection role for EM exposures. To test this, chick embryos (96 h) were exposed to 60 Hz magnetic fields prior to being placed into anoxia. Survival was 39.6% (control), and 68.7% (field-exposed). As a positive control, embryos were heated prior to anoxia (57.6% survival). We conclude that: 1) 60 Hz magnetic field exposures reduce anoxia-induced mortality in chick embryos, comparable to reductions observed following heat stress, and 2) this is a simple and rapid experiment to demonstrate the existence of weak EM field effects. PMID- 9849921 TI - Gender, menstrual cycle, oral contraceptives and red blood cell deformability in healthy adult subjects. AB - Gender, menstrual cycle and oral contraceptives may have influence on mechanical properties of Red Blood Cell (RBC) and particularly on RBC deformability. So cell transit parameters have been assessed by filtration with the Cell Transit Analyser (CTA) for a large healthy adult population (seventy-nine males and one hundred-fifteen females). The CTA provides the distribution of cell transit times of 5000 red blood cells, the mean transit time of the population and different percentiles such as p50, p75, p90 and p95. No effect of oral contraceptives was found. Nevertheless, influence of sex and menstrual cycle were demonstrated. A significant increase of the filtration parameters measured in the female population with respect to the male population and during menstruation, preovulation and post-ovulation periods was observed. During ovulation, the CTA parameters are comparable to the same parameters found in males. PMID- 9849922 TI - The triphasic effects of exercise on blood rheology: which relevance to physiology and pathophysiology? AB - The life-extending effects of regular exercise are related to a decrease in both coronary and peripheral vascular morbidity, associated with some improvements in cardiovascular risk factors. A possible link between the beneficial metabolic and hemodynamic effects of exercise could be blood rheology, which is markedly affected by exercise. We propose here a description of the hemorheological effects of exercise as a triphasic phenomenon. Short-term effects of exercise are an increase in blood viscosity resulting from both fluid shifts and alterations of erythrocyte rheologic properties (rigidity and aggregability). Increased blood lactate, stress, and acute phase play a role in this process. Middle-term effects of regular exercise are a reversal of these acute effects with an increase in blood fluidity, explained by plasma volume expansion (autohemodilution) that lowers both plasma viscosity and hematocrit. Long-term effects further improve blood fluidity, parallel with the classical training-induced hormonal and metabolic alterations. While body composition, blood lipid pattern, and fibrinogen improve (thus decreasing plasma viscosity), erythrocyte metabolic and rheologic properties are modified, with a reduction in aggregability and rigidity. On the whole, these improvements reflect a reversal of the so-called "insulin-resistance syndrome" induced by a sedentary lifestyle. Since impaired blood rheology has been demonstrated to be at risk for vascular diseases, the hemorheologic effects of exercise can be hypothesized to be a mechanism (or at least a marker) of risk reversal. This latter point requires further investigation. The physiological meaning of the triphasic pattern of exercise induced alterations of blood rheology is uncompletely understood, but increased blood fluidity may improve several steps of oxygen transfer to muscle, as clearly demonstrated in hypoxic conditions. Increasing evidence emerges from the literature, that blood fluidity is a physiological determinant of fitness. PMID- 9849923 TI - Haemorheological consequences of hyperglycaemic spike in healthy volunteers and insulin-dependent diabetics. AB - The present study has been designed to examine the role of a hyperglycaemic spike of short duration as a factor possibly involved in haemorheological microcirculatory and (or) haemostatic dysfunctions in ten insulin-dependent diabetes mellitus patients (IDDM) and five healthy volunteers. The hyperglycaemic spike was induced by glucose infusion via GCIIS Biostator. Blood viscosity, erythrocyte aggregation, erythrocyte deformability, plasma viscosity and proteins' levels, cell counts, and transcutaneous oxygen pressure were investigated during normoglycaemic period and at the end of a 1 h hyperglycaemic spike. Hyperglycaemia induced in IDDM patients significant decreases in erythrocyte aggregation, in blood and plasma viscosities and in both fibrinogen and albumin levels. The number of platelets was significantly decreased as a result of the hyperglycaemic spike. There was no significant change induced by the hyperglycaemic spike in healthy volunteers. These findings show that a hyperglycaemic spike of 280 mg/dl (15.6 mmol/l) of short duration in IDDM patients causes numerous significant changes in both blood components and rheological behaviour. These changes were accompanied by a significant decrease in transcutaneous oxygen tension indicating an impaired tissue oxygenation during the hyperglycaemia. PMID- 9849924 TI - A modified Casson equation to characterize blood rheology for hypertension. AB - A modified Casson equation was established, verified satisficatorily for real conditions and suggested for use in clinical hemorheology. It provides 5 Casson parameters, i.e., Casson yield stress, Casson viscosity, the first critical point of shear rate to cause rouleaux to be broken down into smaller units, the second critical point of shear rate to completely disaggregate into individual erythrocytes, the difference between the two critical points, and also gives two series of the 5 above-mentioned Casson parameters, respectively, under original hematocrit and the standardized hematocrit of 45%. This equation not only reveals the apparent abnormalities of hypertensive patients and normalities of the healthy persons, but also reveals hidden abnormalities for patients with either low or high hematocrit. For 20 cases of hypertensive patients with low hematocrit, the 5 Casson parameters have lower values than control, but the modified equation reveals their hidden higher values than control when their hematocrit is standardized to 45%. For 7 cases of polycythemia complicated with hypertension, the 5 Casson parameters are higher at their original hematocrit, and the modified equation reveals the higher parameters than control also when the hematocrit is standardized to 45%. For 8 cases of polycythemia without complications, the 5 Casson parameters are higher at their original hematocrit than control, while the modified equation still reveals their normalities when standardizing their hematocrit to 45%. PMID- 9849925 TI - Haemorheology and the inflammatory process. AB - Many forms of tissue injury, whether due to physical or infectious sources, lead to the inflammatory response. During its course a wide variety of events takes place, including alterations in vascular contraction, in vascular permeability, in leucocyte activity and the initiation of the acute phase response. The last results in a leucocytosis and changes in the concentrations of a number of plasma proteins, including fibrinogen. They all cause alterations in haemodynamics, due in part to alteration in geometric resistance, but also to alterations in viscometric resistance as a result of changes in the haemorheological properties of the blood components locally and systemically. While some of these inflammation-induced changes are useful on a local level and assist in the resolution of the damaging factor and in tissue repair, the systemic haemorheological effects may lead to deleterious haemodynamics. Many of the effects are well documented but two new possibilities arise. One is the potential effects of fibrinogen heterogeneity on blood rheology, and the other the possible effects of leucocyte protease release on the aggregability of the red cell. PMID- 9849926 TI - Plasma thrombomodulin is not increased in familial hypercholesterolemic children with rheological alterations. AB - In order to ascertain whether hemorheological alterations precede the atherosclerotic lesion in familial hypercholesterolemia (FH), we studied the lipid and hemorheological profile of 29 heterozygous FH children (12 males and 17 females), aged 12+/-3 years, and a well-matched control group (CG), and assessed their plasma thrombomodulin level as an early marker of endothelial injury. No differences were found between FH children's plasma thrombomodulin values (31.7+/ 11.2 ng/ml) and those of the CG (27.8+/-15 ng/ml). However, the rheological variables of FH children and the CG were statistically different (p < 0.001) for fibrinogen (Fbg): 266+/-48 mg/dl vs. 205+/-32 mg/dl; erythrocyte aggregation at stasis (EAM0): 4.6+/-1.2 vs. 3.3+/-0.9; erythrocyte aggregation at low shear (EAM1): 7.9+/-1.7 vs. 6.1+/-0.8; and plasma viscosity (PV): 1.18+/-0.03 cP vs. 1.12+/-0.04 cP. Correlations between rheological parameters and lipids were found. The normal values obtained in FH children for plasma thrombomodulin suggest that the hemorheological alterations appear prior to the vascular injury, are in part related to dyslipemia and could contribute to the development of the atherosclerotic process by modifying blood flow conditions. PMID- 9849927 TI - Early rheological and microcirculatory changes in children with type I diabetes mellitus. AB - In order to determine early changes in microcirculation and hemorheological parameters in diabetics, a cross-sectional study was carried out with 273 children with diabetes mellitus type I during their vacation in a state country convalescent home for diabetic children and teenagers in Kaiserslautern. Compared to healthy children, typical changes of hemorheological variables as well as in the microcirculation of the skin and retina are observed in poorly controlled diabetic children. Morphological changes are obvious in capillary areas in form of marked capillary contortions and dilatations of venous branches, rigid erythrocytes, and hyperaggregable thrombocytes. An effort should be undertaken to normalize the pathologically changed parameters of blood fluidity and the microcirculation by an adequate control of blood glucose, and possibly by changes in dietary habits. PMID- 9849929 TI - Red blood cell aggregability in blood. Criteria for scoring the techniques. PMID- 9849928 TI - Molecular rotors as fluorescent probes for biological studies. AB - Molecular rotors, which structure can be 4-(N,N-dimethylamino)-benzene, benzylidene and -cinnamylidene derivatives and, also, coumarine-like compounds, have photophysical characteristics which strongly depend on the environmental parameters (polarity, viscosity, temperature, etc.). In this paper, a basic knowledge on molecular fluorescent rotors will be reminded and two fields of applications using molecular fluorescent rotors as optical sensors will be described: firstly, in polymer and, more particularly to detect the formation of hydrophobic microdomains, in the case of the aggregation of amphiphilic polymers (as models for globular proteins and/or enzymes) and, secondly, in cell biology, especially in liposomes (as models for biological membranes) to follow their thermotropic behavior and in endothelial cells under 3D fluorescence microscopy. PMID- 9849930 TI - Plasma viscosity in nephrotic Nigerians. AB - A simple and reproducible 'syringe viscometer' method was used to determine plasma viscosity, an important haemorheological parameter, in 27 (14 M, 13 F) nephrotic Nigerian patients. The plasma fibrinogen (PFC) and immunoglobulin levels (IgG, IgM, IgA) were also determined in these patients. Similar determinations were carried out in 30 (15 M, 15 F) age- and sex-matched healthy control subjects for comparison. The nephrotic patients had significantly higher relative plasma viscosity (RPV), PFC and IgM levels (2.02+/-0.17 vs. 1.66+/-0.19, 6.48+/-2.59 vs. 2.26+/-0.91 g/l; 316.2+/-169.0 vs. 186.0+/-59.5 IU/ml, p < 0.001, respectively). However, IgG and IgA levels were significantly less (151.1+/-88.7 vs. 205.7+/-77.6 IU/ml, p < 0.05, and 95.8+/-41.5 vs. 127.3+/-31.3 IU/ml, p < 0.01, respectively). In the nephrotic patients, plasma fibrinogen and IgM levels had a significant positive correlation with RPV (r=0.444, p < 0.05, and r=0.452, p < 0.02, respectively). The observed increased plasma viscosity in our nephrotic patients could on the long term predispose them to increased risk for cardiovascular morbidity. PMID- 9849931 TI - Exercise-induced growth hormone secretion and hemorheology during exercise in elite athletes. PMID- 9849932 TI - Helix-helix packing angle preferences for finite helix axes. AB - Recently, James Bowie addressed the question of how to normalize correctly the distribution of observed helix-helix packing angles in proteins (Bowie, Nature Struct. Biol. 4:915-917, 1997). A hitherto unrealized yet significant bias toward crossed packing angles was revealed. However, the derived random reference distribution of packing angles requires that helices have to be assumed as infinite in length. Here, we complement Bowie's analysis by consideration of the more realistic case where helices are of finite length. As a result, the statistical bias toward near perpendicular packings appears to be even stronger. PMID- 9849933 TI - Predicting peptides that bind to MHC molecules using supervised learning of hidden Markov models. AB - The binding of a major histocompatibility complex (MHC) molecule to a peptide originating in an antigen is essential to recognizing antigens in immune systems, and it has proved to be important to use computers to predict the peptides that will bind to an MHC molecule. The purpose of this paper is twofold: First, we propose to apply supervised learning of hidden Markov models (HMMs) to this problem, which can surpass existing methods for the problem of predicting MHC binding peptides. Second, we generate peptides that have high probabilities to bind to a certain MHC molecule, based on our proposed method using peptides binding to MHC molecules as a set of training data. From our experiments, in a type of cross-validation test, the discrimination accuracy of our supervised learning method is usually approximately 2-15% better than those of other methods, including backpropagation neural networks, which have been regarded as the most effective approach to this problem. Furthermore, using an HMM trained for HLA-A2, we present new peptide sequences that are provided with high binding probabilities by the HMM and that are thus expected to bind to HLA-A2 proteins. Peptide sequences not shown in this paper but with rather high binding probabilities can be obtained from the author. PMID- 9849934 TI - A hierarchical method for generating low-energy conformers of a protein-ligand complex. AB - A novel dynamical protocol for finding the low-energy conformations of a protein ligand complex is described. The energy functions examined consist of an empirical force field with four different dielectric screening models; the generalized Born/surface area model also is examined. Application of the method to three complexes of known crystal structure provides insights into the energy functions used for selecting low-energy docked conformations and into the structure of the binding-energy surface. Evidence is presented that the local energy minima of a ligand in a binding site are arranged in a hierarchical fashion. This observation motivates the construction of a hierarchical docking algorithm that substantially enriches the population of ligand conformations close to the crystal conformation. The algorithm is also adapted to permit docking into a flexible binding site and preliminary tests of this method are presented. PMID- 9849935 TI - Energy landscape of a native protein: jumping-among-minima model. AB - We have investigated energy landscape of human lysozyme in its native state by using principal component analysis and a model, jumping-among-minima (JAM) model. These analyses are applied to 1 nsec molecular dynamics trajectory of the protein in water. An assumption embodied in the JAM model allows us to divide protein motions into intra-substate and inter-substate motions. By examining intra substate motions, it is shown that energy surfaces of individual conformational substates are nearly harmonic and mutually similar. As a result of principal component analysis and JAM model analysis, protein motions are shown to consist of three types of collective modes, multiply hierarchical modes, singly hierarchical modes, and harmonic modes. Multiply hierarchical modes, the number of which accounts only for 0.5% of all modes, dominate contributions to total mean-square atomic fluctuation. Inter-substate motions are observed only in a small-dimensional subspace spanned by the axes of multiplyhierarchical and singly hierarchical modes. Inter-substate motions have two notable time components: faster component seen within 200 psec and slower component. The former involves transitions among the conformational substates of the low-level hierarchy, whereas the latter involves transitions of the higher level substates observed along the first four multiply hierarchical modes. We also discuss dependence of the subspace, which contains conformational substates, on time duration of simulation. PMID- 9849936 TI - Patterns of protein-fold usage in eight microbial genomes: a comprehensive structural census. AB - Eight microbial genomes are compared in terms of protein structure. Specifically, yeast, H. influenzae, M. genitalium, M. jannaschii, Synechocystis, M. pneumoniae, H. pylori, and E. coli are compared in terms of patterns of fold usage-whether a given fold occurs in a particular organism. Of the approximately 340 soluble protein folds currently in the structure databank (PDB), 240 occur in at least one of the eight genomes, and 30 are shared amongst all eight. The shared folds are depleted in allhelical structure and enriched in mixed helix-sheet structure compared to the folds in the PDB. The top-10 most common of the shared 30 are enriched in superfolds, uniting many non-homologous sequence families, and are especially similar in overall architecture-eight having helices packed onto a central sheet. They are also very different from the common folds in the PBD, highlighting databank biases. Folds can be ranked in terms of expression as well as genome duplication. In yeast the top-10 most highly expressed folds are considerably different from the most highly duplicated folds. A tree can be constructed grouping genomes in terms of their shared folds. This has a remarkably similar topology to more conventional classifications, based on very different measures of relatedness. Finally, folds of membrane proteins can be analyzed through transmembrane-helix (TM) prediction. All the genomes appear to have similar usage patterns for these folds, with the occurrence of a particular fold falling off rapidly with increasing numbers of TM-elements, according to a "Zipf-like" law. This implies there are no marked preferences for proteins with particular numbers of TM-helices (e.g. 7-TM) in microbial genomes. PMID- 9849938 TI - The slow step of folding of Staphylococcus aureus PC1 beta-lactamase involves the collapse of a surface loop rate limited by the trans to cis isomerization of a non-proline peptide bond. AB - We wished to test the hypothesis that the non proline cis to trans isomerization of the peptide bond at position 167 in the S. aureus beta-lactamase PC1 exerts a significant controlling effect on the folding pathway of this enzyme. The previous data presented in support of this hypothesis could not rule out the effect of factors unrelated to non-proline cis/trans isomerization. We have used the plasmid pET9d to direct soluble overproduction of the S. aureus beta lactamase PC1 and a site-directed mutant (Ile 167 to Pro) in Escherichia coli. Following purification the proteins were subjected to a comparative analysis of the kinetics of unfolding and refolding using the techniques of near- and far-UV circular dichroism spectroscopy and fluorescence spectroscopy in conjunction with "double-jump" experiments. Results show that the fully-unfolded I167P mutant enzyme retains 20% of molecules in a fast-refolding form and that slower refolding molecules fold faster than the recombinant wild-type enzyme. The final stage of folding involves folding of the omega-loop into a conformation essential for enzymatic activity. In support of the original hypothesis, the folding of this omega-loop is rate limited by the isomerization of the Glu 166-Ile 167 peptide bond. PMID- 9849937 TI - Modelling repressor proteins docking to DNA. AB - The docking of repressor proteins to DNA starting from the unbound protein and model-built DNA coordinates is modeled computationally. The approach was evaluated on eight repressor/DNA complexes that employed different modes for protein/ DNA recognition. The global search is based on a protein-protein docking algorithm that evaluates shape and electrostatic complementarity, which was modified to consider the importance of electrostatic features in DNA-protein recognition. Complexes were then ranked by an empirical score for the observed amino acid /nucleotide pairings (i.e., protein-DNA pair potentials) derived from a database of 20 protein/ DNA complexes. A good prediction had at least 65% of the correct contacts modeled. This approach was able to identify a good solution at rank four or better for three out of the eight complexes. Predicted complexes were filtered by a distance constraint based on experimental data defining the DNA footprint. This improved coverage to four out of eight complexes having a good model at rank four or better. The additional use of amino acid mutagenesis and phylogenetic data defining residues on the repressor resulted in between 2 and 27 models that would have to be examined to find a good solution for seven of the eight test systems. This study shows that starting with unbound coordinates one can predict three-dimensional models for protein/DNA complexes that do not involve gross conformational changes on association. PMID- 9849939 TI - Homology modeling of an RNP domain from a human RNA-binding protein: Homology constrained energy optimization provides a criterion for distinguishing potential sequence alignments. AB - We have recently described an automated approach for homology modeling using restrained molecular dynamics and simulated annealing procedures (Li et al, Protein Sci., 6:956-970,1997). We have employed this approach for constructing a homology model of the putative RNA-binding domain of the human RNA-binding protein with multiple splice sites (RBP-MS). The regions of RBP-MS which are homologous to the template protein snRNP U1A were constrained by "homology distance constraints," while the conformation of the non-homologous regions were defined only by a potential energy function. A full energy function without explicit solvent was employed to ensure that the calculated structures have good conformational energies and are physically reasonable. The effects of mis alignment of the unknown and the template sequences were also explored in order to determine the feasibility of this homology modeling method for distinguishing possible sequence alignments based on considerations of the resulting conformational energies of modeled structures. Differences in the alignments of the unknown and the template sequences result in significant differences in the conformational energies of the calculated homology models. These results suggest that conformational energies and residual constraint violations in these homology constrained simulated annealing calculations can be used as criteria to distinguish between correct and incorrect sequence alignments and chain folds. PMID- 9849940 TI - Structural basis of increased resistance to thermal denaturation induced by single amino acid substitution in the sequence of beta-glucosidase A from Bacillus polymyxa. AB - The increasing development of the biotechnology industry demands the design of enzymes suitable to be used in conditions that often require broad resistance against adverse conditions. beta-glucosidase A from Bacillus polymyxa is an interesting model for studies of protein engineering. This is a well characterized enzyme, belonging to glycosyl hydrolase family 1. Its natural substrate is cellobiose, but is also active against various artificial substrates. In its native state has an octameric structure. Its subunit conserves the general (alpha/beta)8 barrel topology of its family, with the active site being in a cavity defined along the axis of the barrel. Using random-mutagenesis, we have identified several mutations enhancing its stability and it was found that one them, the E96K substitution, involved structural changes. The crystal structure of this mutant has been determined by X-ray diffraction and compared with the native structure. The only difference founded between both structures is a new ion pair linking Lys96 introduced at the N-terminus of helix alpha2, to Asp28, located in one of the loops surrounding the active-site cavity. The new ion pair binds two segments of the chain that are distant in sequence and, therefore, this favorable interaction must exert a determinant influence in stabilizing the tertiary structure. Furthermore, analysis of the crystallographic isotropic temperature factors reveals that, as a direct consequence of the introduced ion pair, an unexpected decreased mobility of secondary structure units of the barrel which are proximal to the site of mutation is observed. However, this effect is observed only in the surrounding of one of the partners forming the salt bridge and not around the other. These results show that far reaching effects can be achieved by a single amino acid replacement within the protein structure. Consequently, the identification and combination of a few single substitutions affecting stability may be sufficient to obtain a highly resistant enzyme, suitable to be used under extreme conditions. PMID- 9849941 TI - Surface properties of adipocyte lipid-binding protein: Response to lipid binding, and comparison with homologous proteins. AB - Adipocyte lipid-binding protein (ALBP) is one of a family of intracellular lipid binding proteins (iLBPs) that bind fatty acids, retinoids, and other hydrophobic ligands. The different members of this family exhibit a highly conserved three dimensional structure; and where structures have been determined both with (holo) and without (apo) bound lipid, observed conformational changes are extremely small (Banaszak, et al., 1994, Adv. Prot. Chem. 45, 89; Bernlohr, et al., 1997, Annu. Rev. Nutr. 17, 277). We have examined the electrostatic, hydrophobic, and water accessible surfaces of ALBP in the apo form and of holo forms with a variety of bound ligands. These calculations reveal a number of previously unrecognized changes between apo and holo ALBP, including: 1) an increase in the overall protein surface area when ligand binds, 2) expansion of the binding cavity when ligand is bound, 3) clustering of individual residue exposure increases in the area surrounding the proposed ligand entry portal, and 4) ligand binding dependent variation in the topology of the electrostatic potential in the area surrounding the ligand entry portal. These focused analyses of the crystallographic structures thus reveal a number of subtle but consistent conformational and surface changes that might serve as markers for differential targeting of protein-lipid complexes within the cell. Most changes are consistent from ligand to ligand, however there are some ligand-specific changes. Comparable calculations with intestinal fatty-acid-binding protein and other vertebrate iLBPs show differences in the electrostatic topology, hydrophobic topology, and in localized changes in solvent exposure near the ligand entry portal. These results provide a basis toward understanding the functional and mechanistic differences among these highly structurally homologous proteins. Further, they suggest that iLBPs from different tissues exhibit one of two predominant end state structural distributions of the ligand entry portal. PMID- 9849942 TI - Kinetics and interaction studies between cytochrome c3 and Fe-only hydrogenase from Desulfovibrio vulgaris Hildenborough. AB - Hydrogenases from Desulfovibrio are found to catalyze hydrogen uptake with low potential multiheme cytochromes, such as cytochrome c3, acting as acceptors. The production of Fe-only hydrogenase from Desulfovibrio vulgaris Hildenborough was improved with respect to the growth phase and media to determine the best large scale bacteria growth conditions. The interaction and electron transfer from Fe only hydrogenase to multiheme cytochrome has been studied in detail by both BLAcore and steady-state measurements. The electron transfer between [Fe] hydrogenase and cytochrome c3 appears to be a cooperative phenomenon (h = 1.37). This behavior could be related to the conductivity properties of multihemic cytochromes. An apparent dissociation constant was determined (2 x 10(-7) M). The importance of the cooperativity for contrasting models proposed to describe the functional role of the hydrogenase/cytochrome c3 complex is discussed. Presently, the only determined structure is from [NiFe] hydrogenase and there are no obvious similarities between [NiFe] and [Fe] hydrogenase. Furthermore, no crystallographic data are available concerning [Fe] hydrogenase. The first results on crystallization and X-ray crystallography are reported. PMID- 9849943 TI - Structural models of the bovine papillomavirus E5 protein. AB - The bovine papillomavirus E5 protein is thought to be a type II integral membrane protein that exists as a disulfide-linked homodimer in transformed cells. Polarized-infrared measurements show that the E5 dimer in membrane bilayers is largely alpha-helical and has a transmembrane orientation. Computational searches of helix-helix conformations reveal two possible low-energy dimer structures. Correlation of these results with previous mutagenesis studies on the E5 protein suggests how the E5 dimer may serve as a molecular scaffold for dimerization and ligand-independent activation of the PDGF-beta receptor. We propose that on each face of the E5 dimer a PDGF-beta receptor molecule interacts directly with Gln17 from one E5 monomer and with Asp33 from the other E5 monomer. This model accounts for the requirement of Gln17 and Asp33 for complex formation and explains genetic results that dimerization of the E5 protein is essential for cell transformation. PMID- 9849944 TI - Grading and hemodynamic follow-up study of arteriovenous malformations with transcranial Doppler ultrasonography. AB - This study presents and tests the clinical validity of a hemodynamic grading system that depends on noninvasive transcranial Doppler ultrasonographic parameters. The suggested transcranial Doppler-based grading system was compared with the Spetzler-Martin anatomic grading for prognosticative validity and clinical dependability. We concluded the following: (1) The pulsatility index was shown to be a more dependable transcranial Doppler parameter in the clinical evaluation of an arteriovenous malformation because of two reasons: preoperative pulsatility index findings inversely correlated with arteriovenous malformation volume, and the pulsatility index returned to normal values before the mean blood flow velocity did. Therefore, hemodynamic arteriovenous malformation grading can be based on the pulsatility index. (2) A transcranial Doppler-based hemodynamic arteriovenous malformation grading system correlated highly with the Spetzler Martin grading in predicting postoperative neurologic deficits and adverse radiologic findings. (3) The presented grading system may contribute to the standardization and quantification of the hemodynamic changes during multidisciplinary management of arteriovenous malformations. PMID- 9849945 TI - Ultrasonographically guided percutaneous biopsy of gastric, small bowel, and colonic abnormalities: efficacy and safety. AB - The purposes of this study were to evaluate the efficacy of sonographically guided percutaneous biopsy of gastric and bowel lesions and to document any associated complications. A retrospective review of all patients undergoing ultrasonographically guided biopsy of gastric, small bowel, or colonic lesions was performed. Cases were evaluated for size of lesion, location, efficacy in obtaining a diagnostic specimen, and any procedure-related complications. A total of 46 biopsies were performed in 44 patients over a 6 year period. Fifteen gastric, four duodenal, seven small bowel, 16 right colonic, and two left colonic lesions were identified. Histologic diagnosis included 20 adenocarcinomas, seven lymphomas, 10 mesenchymal tumors, and two cases of candidal enteritis. In 18 (41%) patients the initial fine-needle aspirate was negative, although 10 of these patients subsequently underwent ultrasonographically guided core biopsy with a diagnosis of neoplasm established in all 10. Two patients with an initial negative result on fine-needle aspiration underwent repeat aspiration, demonstrating malignancy. Two additional false-negative aspirates were obtained; these patients did not undergo repeat biopsy but were subsequently found to have lymphoma. Two of the patients with aspirates that were negative for malignancy were believed to have inflammatory changes secondary to pancreatitis. One patient was lost to follow-up evaluation. No complications were recorded. Percutaneous fine-needle aspiration of gastric and bowel lesions is a simple and safe procedure and may be particularly useful with poor candidates for endoscopy or for lesions that are difficult or impossible to reach endoscopically. Diagnostic yield is high, particularly in documenting malignancy, although in suspected stromal tumors a core biopsy often is required. PMID- 9849946 TI - Bladder distention and pyelectasis in the male fetus: causes, comparisons, and contrasts. AB - The objective of this paper was to determine if prenatal sonographic findings can accurately differentiate between the causes of bladder distention and pyelectasis in the male fetus. Twenty-one cases were evaluated for the presence of oligohydramnios, posterior urethral dilation, bladder wall thickening, urachal patency, cortical thinning, cortical cysts, and increased renal echogenicity. Postnatal diagnosis included posterior urethral valves (10 cases), prune belly syndrome (four cases), vesicoureteral reflux (four cases), left ureterovesical junction obstruction (one case), and nonrefluxing, nonobstructive megacystis megaureter (two cases). Oligohydramnios was present in eight of 10 cases of posterior urethral valves and in one of four cases of prune belly syndrome. A dilated posterior urethra was noted in seven of 10 cases of posterior urethral valves and transiently in two of four cases of prune belly syndrome. Bladder wall thickening developed in all cases of posterior urethral valves and was noted in two of four patients with prune belly syndrome. A patent urachus likewise was identified in two of four cases of prune belly syndrome. The presence of oligohydramnios, progressive bladder wall thickening, and dilated posterior urethra was most suggestive of posterior urethral valves, whereas the presence of a patent urachus was most suggestive of prune belly syndrome. The presence of pyelectasis and megacystis without additional amniotic fluid, bladder, urethral, or renal abnormalities was most suggestive of vesicoureteral reflux, ureterovesical junction obstruction, or nonrefluxing, nonobstructive megacystis megaureter. Owing to the overlap and evolution of these findings, close follow-up evaluation is recommended. PMID- 9849947 TI - A 17 year old patient with twin gestation hospitalized because of premature rupture of the membranes. PMID- 9849948 TI - Comparison of power Doppler and B-scan sonography for renal imaging using a sonographic contrast agent. AB - The goal of this study was to evaluate the relative performance of power Doppler and B-scan imaging modes in detecting vascular perfusion changes resulting from injection of a contrast agent. To allow this comparison the imaging plane and the contrast agent injection must be the same for both modes. We achieved this by using a rigid transducer holder and simultaneously recording power Doppler and B scan images on separate videotapes. The kidneys of five adult beagles were scanned to allow a comparison of how power Doppler and B-scan imaging methods monitor changes during the injection of 0.1 ml/kg of a contrast agent, EchoGen emulsion (Sonus Pharmaceuticals, Bothell, WA). The changes in the images were assessed qualitatively by three radiologists and quantitatively using a custom designed image analysis software. All of the radiologists agreed that no visually detectable changes occurred in B-scan images but that significant changes could be observed in power Doppler images. Image analysis also indicated a difference between power Doppler and B-scan images. The change in mean color level of power Doppler images could be displayed as an indicator dilution curve with a peak enhancement of 46 +/- 16 above the preinjection value. The time at which mean color level peaked was 18 +/- 13 s. The mean color level returned to half of the peak value by 69 +/- 42 s and returned to the preinjection baseline value by 148 +/- 73 s. Conversely, B-scan images showed statistically insignificant changes, and time measurements could not be made. By all measures used to evaluate images, power Doppler imaging had a greater sensitivity in detecting changes resulting from contrast agent injection than B-scan imaging. This finding indicates that power Doppler imaging of contrast agent injections can be used to map regional differences in flow as well as quantitative measurements of a contrast agent's transit time and has the potential to assess kidney abnormalities associated with renal blood flow. PMID- 9849949 TI - Volumetric measurements of the brachial and carotid arteries in dialysis grafts: is there evidence of a subclavian-carotid artery steal syndrome? AB - Dialysis grafts may lead to major hyperperfusion in the graft arm and to hypertrophic, hypervolemic cardiomyopathy. No data have been published concerning the impact of dialysis grafts on the cerebral perfusion in relation to a potential carotid steal syndrome, possibly causing neurologic or neuropsychologic symptoms. In 30 patients (32 to 74 years old) with dialysis grafts we studied the following hemodynamic parameters in the brachial and common carotid arteries bilaterally: Flow velocities (spectral Doppler sonography), diameter (B-mode) and volume flow (color M-mode) with a color duplex system (Philips P700). Volume flow in the brachial arteries of the graft arm averaged 1032 ml/min (range, 158 to 2854 ml/min) as compared to 42 ml/min (range, 15 to 108 ml/min) in the nongraft arm. Almost identical volume flow data could be seen in both common carotid arteries (418 versus 421 ml/min) with no evidence of reduced flow in the carotid arteries in patients with high flow in the graft arm. A tendency toward higher volume flow in both carotid arteries in patients with high volume flow in the graft arm was noted. We found no evidence of shunt-induced cerebral hypoperfusion. Cerebral autoregulation appears to be patent even with high brachial artery shunt volume. PMID- 9849950 TI - Frequency and nature of structural anomalies in fetuses with single umbilical arteries. AB - Published studies differ concerning the rate of anomalies occurring in the presence of a single umbilical artery and the significance of the single umbilical artery as an isolated sonographic finding. We assessed the frequency, nature, and sonographic detection of structural anomalies in fetuses with a single umbilical artery. We identified all cases in which prenatal sonography diagnosed a single umbilical artery. Cases were excluded if postnatal physical or pathologic examination demonstrated a three-vessel cord, yielding a study population of 167 cases. For each case, we recorded the gestational age at diagnosis of single umbilical artery and the findings of the sonographic fetal anatomic survey. We recorded postnatal clinical and pathologic information when available. Gestational age at time of diagnosis ranged from 16.8 to 41.1 weeks (mean, 29.2 +/- 6.5 weeks). Twenty of the 167 fetuses (12%) were twins, and the remainder were singletons. Among 118 cases with postnatal information, 37 (31%) had structural abnormalities, often involving multiple organs. The most common organ systems involved were the heart (19 cases) and the gastrointestinal (14 cases) and central nervous systems (nine cases). Five of the anomalous fetuses had abnormal karyotypes. The sonographic survey was abnormal in 31 of the 37 anomalous fetuses (84%). Among 85 cases with apparently isolated single umbilical artery at sonography and known fetal outcome, six (7%) proved to be anomalous at birth. We had two sonographic false-positive results (mild hydronephrosis, suspected skeletal dysplasia). In summary, approximately one third of fetuses with single umbilical artery have structural anomalies, most often cardiac. Even when the single umbilical artery is an apparently isolated sonographic finding, the likelihood that the neonate will prove to have structural anomalies is considerable (7% in our series). PMID- 9849951 TI - Superior mesenteric artery blood flow velocimetry: cross-sectional Doppler sonographic study in normal fetuses. AB - Our objectives were to describe the flow velocity waveform of the fetal superior mesenteric artery and to establish a nomogram for its pulsatility index. In a cross-sectional study using color Doppler ultrasonography, superior mesenteric artery flow velocimetry was investigated prospectively in 96 healthy fetuses of between 14 and 37 weeks of gestation. In normal fetuses the pulsatility index measurements showed a slight but insignificant increase over the course of gestation (r = 0.26; P > 0.5). The lowest mean +/- standard deviation for pulsatility index in the superior mesenteric artery was 1.86 +/- 0.45 (95% confidence interval 1.67-2.06), recorded between 18 and 21 weeks' gestation. Thereafter the pulsatility index increased to 1.94 +/- 6.4 (95% confidence interval 1.74-2.8) at 22 to 25 weeks, and from weeks 26 to 29 it increased to 2.18 +/- 0.52 (95% confidence interval 1.91-2.46). During the third trimester and at term, the mean pulsatility index of 2.23 +/- 0.32 (95% confidence interval 1.43-3.03) did not change significantly with gestational age. In normal fetuses, except for the early stages, a relatively stable vascular resistance of the intestinal circulation was found. The application of this nomogram in clinical practice may facilitate evaluation of intestinal perfusion in compromised fetuses with blood flow centralization. PMID- 9849952 TI - Persistent right umbilical vein in trisomy 18: sonographic observation. PMID- 9849953 TI - Dissecting aneurysm of the superior mesenteric artery: flow dynamics by color Doppler sonography. PMID- 9849954 TI - Intratesticular leiomyoma of the body of the testis. PMID- 9849955 TI - AIUM technical bulletin--Color duplex imaging for graft surveillance of the autologous vein graft. American Institute of Ultrasound in Medicine. PMID- 9849956 TI - The bone morphogenetic protein 15 gene is X-linked and expressed in oocytes. AB - We have taken advantage of the sequence relationships among the bone morphogenetic proteins (BMPs) to identify the mouse Bmp15 and human BMP15 genes. The 392-amino acid prepropeptides encoded by these BMP genes exhibit significant homology to each other, although the 70% identity observed between the 125-amino acid mature peptides is considerably lower than that seen in comparisons of other mouse and human orthologs. Both genes share a common structural organization and encode mature peptides that lack the cysteine residue normally involved in the formation of a covalent dimer. In addition, mouse Bmp15 and human BMP15 map to conserved syntenic regions on the X chromosome. We demonstrate, through a combination of Northern blot and in situ hybridization analyses, that mouse Bmp15 is expressed specifically in the oocyte beginning at the one-layer primary follicle stage and continuing through ovulation. Interestingly, BMP-15 is most closely related to and shares a coincident expression pattern with the mouse growth/differentiation factor 9 (GDF-9) gene that is essential for female fertility. Our findings will be important for defining the role of BMP-15 in follicular development. PMID- 9849957 TI - Agonist-induced endocytosis and recycling of the gonadotropin-releasing hormone receptor: effect of beta-arrestin on internalization kinetics. AB - This study examined the dynamics of endocytotic and recycling events associated with the GnRH receptor, a unique G protein-coupled receptor (GPCR) without the intracellular carboxyl-terminal tail, after agonist stimulation, and investigated the role of beta-arrestin in this process. Subcellular location of fluorescently labeled epitope-tagged GnRH receptors stably expressed in HEK 293 cells was monitored by confocal microscopy, and the receptor/ligand internalization process was quantified using radioligand binding and ELISA. Agonist stimulation resulted in reversible receptor redistribution from the plasma membrane into the cytoplasmic compartment, and colocalization of internalized GnRH receptors with transferrin receptors was observed. Internalization experiments for the GnRH receptor and another GPCR possessing a carboxy-terminal tail, the TRH receptor, showed that the rate of internalization for the GnRH receptor was much slower than for the TRH receptor when expressed in both HEK 293 and COS-7 cells. TRH receptor internalization could be substantially increased by coexpression with beta-arrestin in COS-7 cells, while GnRH receptor internalization was not affected by coexpression with beta-arrestin in either cell type. Coexpression of the GnRH receptor with the dominant negative beta-arrestin (319-418) mutant did not affect its ability to internalize, and activated GnRH receptors did not induce time-dependent redistribution of beta-arrestin/green fluorescent protein to the plasma membrane. However, the beta-arrestin mutant impaired the internalization of the TRH receptor, and activated TRH receptors induced the beta arrestin/green fluorescent protein translocation. This study demonstrates that, despite having no intracellular carboxy-terminal tail, the GnRH receptor undergoes agonist-stimulated internalization displaying distinctive characteristics described for other GPCRs that internalize via a clathrin dependent mechanism and recycle through an acidified endosomal compartment. However, our data indicate that the GnRH receptor may utilize a beta-arrestin independent endocytotic pathway. PMID- 9849958 TI - Characterization of two LGR genes homologous to gonadotropin and thyrotropin receptors with extracellular leucine-rich repeats and a G protein-coupled, seven transmembrane region. AB - The receptors for LH, FSH, and TSH belong to the large G protein-coupled, seven transmembrane (TM) protein family and are unique in having a large N-terminal extracellular (ecto-) domain containing leucine-rich repeats important for interaction with the glycoprotein ligands. We have identified two new leucine rich repeat-containing, G protein-coupled receptors and named them as LGR4 and LGR5, respectively. The ectodomains of both receptors contain 17 leucine-rich repeats together with N- and C-terminal flanking cysteine-rich sequences, compared with 9 repeats found in known glycoprotein hormone receptors. The leucine-rich repeats in LGR4 and LGR5 are arrays of 24 amino acids showing similarity to repeats found in the acid labile subunit of the insulin-like growth factor (IGF)/IGF binding protein complexes as well as slit, decorin, and Toll proteins. The TM region and the junction between ectodomain and TM 1 are highly conserved in LGR4, LGR5, and seven other LGRs from sea anemone, fly, nematode, mollusk, and mammal, suggesting their common evolutionary origin. In contrast to the restricted tissue expression of gonadotropin and TSH receptors in gonads and thyroid, respectively, LGR4 is expressed in diverse tissues including ovary, testis, adrenal, placenta, thymus, spinal cord, and thyroid, whereas LGR5 is found in muscle, placenta, spinal cord, and brain. Hybridization analysis of genomic DNA indicated that LGR4 and LGR5 genes are conserved in mammals. Comparison of overall amino acid sequences indicated that LGR4 and LGR5 are closely related to each other but diverge, during evolution, from the homologous receptor found in snail and the mammalian glycoprotein hormone receptors. The identification and characterization of new members of the LGR subfamily of receptor genes not only allow future isolation of their ligands and understanding of their physiological roles but also reveal the evolutionary relationship of G protein-coupled receptors with leucine-rich repeats. PMID- 9849959 TI - Identification of phosphorylation sites in the G protein-coupled receptor for parathyroid hormone. Receptor phosphorylation is not required for agonist-induced internalization. AB - In some G protein-coupled receptors (GPCRs), agonist-dependent phosphorylation by specific GPCR kinases (GRKs) is an important mediator of receptor desensitization and endocytosis. Phosphorylation and the subsequent events that it triggers, such as arrestin binding, have been suggested to be regulatory mechanisms for a wide variety of GPCRs. In the present study, we investigated whether agonist-induced phosphorylation of the PTH receptor, a class II GPCR, also regulates receptor internalization. Upon agonist stimulation, the PTH receptor was exclusively phosphorylated on serine residues. Phosphoamino acid analysis of a number of receptor mutants in which individual serine residues had been replaced by threonine identified serine residues in positions 485, 486, and 489 of the cytoplasmic tail as sites of phosphorylation after agonist treatment. When serine residues at positions 483, 485, 486, 489, 495, and 498 were simultaneously replaced by alanine residues, the PTH receptor was no longer phosphorylated either basally or in response to PTH. The substitution of these serine residues by alanine affected neither the number of receptors expressed on the cell surface nor the ability of the receptor to signal via Gs. Overexpression of GRK2, but not GRK3, enhanced PTH-stimulated receptor phosphorylation, and this phosphorylation was abolished by alanine mutagenesis of residues 483, 485, 486, 489, 495, and 498. Thus, phosphorylation of the PTH receptor by the endogenous kinase in HEK 293 cells occurs on the same residues targeted by overexpressed GRK2. Strikingly, the rate and extent of PTH-stimulated internalization of mutated PTH receptors lacking phosphorylation sites were identical to that observed for the wild-type PTH receptor. Moreover, overexpressed GRK2, while enhancing the phosphorylation of the wild-type PTH receptor, had no affect on the rate or extent of receptor internalization in response to PTH. Thus, the agonist-occupied PTH receptor is phosphorylated by a kinase similar or identical to GRK2 in HEK-293 cells, but this phosphorylation is not requisite for efficient receptor endocytosis. PMID- 9849960 TI - Certain activating mutations within helix 6 of the human luteinizing hormone receptor may be explained by alterations that allow transmembrane regions to activate Gs. AB - Male-limited gonadotropin-independent precocious puberty (MPP) is frequently associated with mutations of the human LH/CG receptor (hLHR) that result in constitutively active hLHRs. Many such activating mutations have been identified in transmembrane 6 of the hLHR, with the substitution of Asp-578 being the most frequently observed mutation. Mutagenesis of a transmembrane helix of a G protein coupled receptor can cause local alterations in the conformation near the mutated residue, allosteric changes elsewhere in the protein, and/or changes in the interhelical packing of the receptor. Therefore, while it has been hypothesized that activation of the receptor by mutations of Asp-578 may arise via alterations in the interactions of helix 6 with other transmembrane helices and/or by allosterically altering the conformation of the third intracellular loop, it has not been possible to ascertain the role of the sixth transmembrane helix per se in activating Gs in the mutated full-length receptor. Recently, however, we have shown that a peptide KMAILIFT, corresponding to the juxtacytoplasmic portion of helix 6 of the hLHR, is capable of activating Gs. These results suggest that helix 6 itself can directly interact with Gs. Importantly, the KMAILIFT peptide did not include Asp-578, which lies just C-terminal to this sequence. We show herein that a peptide extended to include Asp-578 (KMAILIFTDFT) is a poor activator of Gs. However, if the peptide is synthesized with the aspartate replaced with either a glycine or tyrosine, substitutions that are found in some patients with MPP, these peptides have Gs-stimulating activity. Additionally, a transmembrane 6 peptide with the substitution of Ile-575 with leucine, another mutation found in MPP, mimicked the activating effects of this mutation in the full-length receptor. The ability of peptides in which Asp-578 or Ile-575 is substituted to mimic the activating effects of these mutations in the full-length receptor suggests that the sixth transmembrane helix represents a site for direct interaction with Gs. In addition to the stimulatory effects of transmembrane 6 peptides, peptides corresponding to the juxtacytoplasmic portions of the fourth, fifth, and seventh helices were also able to stimulate Gs. These results are consistent with the hypothesis that the transmembrane helices may form a pocket for interaction with Gs and that constitutive activation of the hLHR may involve the opening of the pocket formed by these helices, thus exposing Gs-binding sites on these helices. PMID- 9849962 TI - A role for guanyl nucleotide-binding regulatory protein beta- and gamma-subunits in the expression of the adrenocorticotropin receptor. AB - Mutant Y1 mouse adrenocortical tumor cells, isolated on the basis of their resistance to the growth-inhibitory effects of forskolin, arise from single mutational events. These mutants present complex phenotypes in which the activity of Gbeta/gamma is impaired, ACTH receptor gene expression is markedly diminished, and ACTH-responsive adenylyl cyclase activity is lost. In this study, we have tested the hypothesis that the impairment in Gbeta/gamma activity is responsible for the loss of ACTH receptor gene expression and ACTH-responsive adenylyl cyclase activity. Transfection of one of the mutant clones with expression vectors encoding either Gbeta1 or Gbeta2 together with Ggamma2 increased ACTH receptor expression and restored ACTH-responsive adenylyl cyclase activity. Interestingly, either Gbeta2 or Ggamma2 alone was effective. These results thus support the hypothesis that the impairment in Gbeta/gamma activity is responsible for the loss of ACTH receptor expression. A luciferase reporter plasmid driven by the proximal promoter region of the mouse ACTH receptor gene was expressed poorly in the mutants compared with parental Y1 cells, suggesting that the Gbeta/gamma defect compromised transcriptional activity at the proximal promoter region of the ACTH receptor gene. PMID- 9849961 TI - Insulin receptor substrate-1 and phosphatidylinositol 3-kinase regulate extracellular signal-regulated kinase-dependent and -independent signaling pathways during myogenic differentiation. AB - Activation of the insulin-like growth factor (IGF) autocrine loop is required for myogenic differentiation and results in sustained activation of extracellular signal-regulated kinases-1 and -2 (ERK-1 and -2). We show here that insulin receptor substrate-1 (IRS-1) phosphorylation on tyrosine and serine residues and association with phosphatidylinositol 3-kinase (PI 3-kinase) are also associated with IGF-dependent myogenic differentiation. Down-regulation of IRS-1 is linked to its serine phosphorylation dependent on PI 3-kinase activity and appears required for differentiation to occur, as IRS-1 is not modified and continues to accumulate in a nondifferentiating myoblast cell line. Furthermore, inhibition of PI 3-kinase activity with LY294002 blocks differentiation, as demonstrated by inhibition of myogenin and myosin heavy chain expression and ERK activation. Blocking the Raf/MEK/ERK cascade with PD98059 does not block myogenic differentiation; however, myotubes do not survive. Thus, PI 3-kinase, in association with IRS-1, is involved in an ERK-independent signaling pathway in myoblasts required for IGF-dependent myogenic differentiation and in inducing sustained activation of ERKs necessary for later stages of differentiation. PMID- 9849963 TI - A novel natural mutation in the thyroid hormone receptor defines a dual functional domain that exchanges nuclear receptor corepressors and coactivators. AB - In a patient with severe resistance to thyroid hormone (RTH), we found a novel mutation (leucine to serine in codon 454, L454S) of the thyroid hormone receptor beta. This mutation is in the ligand-dependent transactivation domain that has been shown to interact with transcriptional coactivators (CoAs). The mutant protein binds T3, but its ability to activate transcription of a positively regulated gene (TRE-tk-Luc), and to repress a negatively regulated gene (TSHalpha Luc), is markedly impaired. As anticipated from its location, the L454S mutant interacts weakly with CoAs, such as SRC1 and glucocorticoid receptor interacting protein 1 (GRIP1) in gel mobility shift assays and in mammalian two-hybrid assays, even in the presence of the maximal dose of T3. In contrast, in the absence of T3, the L454S mutant interacts much more strongly with nuclear receptor corepressor (NCoR) than does the wild-type receptor, and the T3 dependent release of NCoR is markedly impaired. By comparison, the NCoR interaction and T3-dependent dissociation of an adjacent AF-2 domain mutant (E457A) are normal. These findings reveal that the Leu 454 is involved directly, or indirectly, in the release of corepressors (CoRs) as well as in the recruitment of CoAs. The strong interaction with NCoR at a physiological concentration of T3 results in constitutive activation of the TSH genes as well as constitutive silencing of positively regulated genes. When the dominant negative effect was examined among various mutants, it correlated surprisingly well with the potency of NCoR binding but not with the degree of impairment in CoA binding. These findings suggest that the defective release of NCoRs, along with retained dimerization and DNA binding, are critical features for the inhibitory action of mutant thyroid hormone receptors. These studies also suggest that helix 12 of the thyroid hormone receptor acts as a dual functional domain. After the binding of T3, its conformation changes, causing the disruption of CoR binding and the recruitment of CoAs. PMID- 9849964 TI - Heat shock protein 90-dependent (geldanamycin-inhibited) movement of the glucocorticoid receptor through the cytoplasm to the nucleus requires intact cytoskeleton. AB - We use here a chimera of the green fluorescent protein (GFP) and the glucocorticoid receptor (GR) to test the notion that the protein chaperone heat shock protein-90 (hsp90) is required for steroid-dependent translocation of the receptor through the cytoplasm along cytoskeletal tracks. The GFP-GR fusion protein undergoes steroid-mediated translocation from the cytoplasm to the nucleus, where it is transcriptionally active. Treatment of 3T3 cells containing steroid-bound GFP-GR with geldanamycin, a benzoquinone ansamycin that binds to hsp90 and disrupts its function, inhibits dexamethasone-dependent translocation from the cytoplasm to the nucleus. The t1/2 for translocation in the absence of geldanamycin is approximately 5 min, and the t1/2 in the presence of geldanamycin is approximately 45 min. In cells treated for 1 h with the cytoskeletal disrupting agents colcemid, cytochalasin D, and beta,beta'-iminodipropionitrile to completely disrupt the microtubule, microfilament, and intermediate filament networks, respectively, the GFP-GR still translocates rapidly to the nucleus in a strictly dexamethasone-dependent manner but translocation is no longer affected by geldanamycin. After withdrawal of the cytoskeletal disrupting agents for 3 h, normal cytoskeletal architecture is restored, and geldanamycin inhibition of dexamethasone-dependent GFP-GR translocation is restored. We suggest that in cells without an intact cytoskeletal system, the GFP-GR moves through the cytoplasm by diffusion. However, under physiological conditions in which the cytoskeleton is intact, diffusion is limited, and the GFP-GR utilizes a movement machinery that is dependent upon hsp90 chaperone activity. In contrast to the GR, GFP-STAT5B, a signaling protein that is not complexed with hsp90, undergoes GH dependent translocation to the nucleus in a manner that is not dependent upon hsp90 chaperone activity. PMID- 9849965 TI - Agonist and antagonists induce homodimerization and mixed ligand heterodimerization of human progesterone receptors in vivo by a mammalian two hybrid assay. AB - This study utilizes the mammalian two-hybrid system to examine the role of ligand in the dimerization of human progesterone receptor (hPR). The GAL4 DNA-binding domain and the herpes simplex virus VP16 transactivation domain were fused to the amino terminus of full-length hPR (both the A and B isoforms) to produce chimeric proteins. PR dimerization was detected by the ability of cotransfected GAL4/PR and VP16/PR chimeras in COS cells to induce expression of a reporter gene under the control of GAL4-binding sites (pG5CAT). Hormone agonist-dependent interactions were observed between the two like isoforms of PR (A-A and B-B) and between PR-A and PR-B (A-B), indicating that hormone can stimulate the formation of the three possible dimeric forms of PR within cells. In contrast, neither type I (ZK98299) nor type II (RU486, ZK112993) progestin antagonists stimulated interaction between these same hybrid PR proteins. However, activation of the VP16/PR chimera by antagonists on a progesterone response element-controlled reporter gene (DHRE-E1b-CAT) was only a fraction (4-13%) of that stimulated by agonist R5020. One possibility for the failure to detect an induction in the two hybrid assay is antagonist-induced repression of the activity of the VP16/PR fusion protein rather than a failure of antagonists to stimulate interaction between the hybrid proteins. To test this idea, an UP-1 carboxyl-terminal truncation mutant of PR was used to construct the two-hybrid proteins. PR-UP-1 selectively binds antagonists, but not agonists, and is fully activated in response to antagonists. Both types of progestin antagonists stimulated interactions between GAL4/PR(UP-1) and VP16/PR(UP-1) hybrid proteins, indicating that antagonists are capable of stimulating PR dimerization in cells and do not function by disrupting or preventing dimerization. To determine whether PR bound to an antagonist can dimerize in whole cells with PR bound to agonist, GAL4/PR(UP 1) was paired in the two-hybrid assay with a VP16/PR fusion protein harboring a point mutation in PR at amino acid 722 (Gly-Cys) that specifically binds progestin agonist but not antagonist. Neither R5020 nor RU486 alone stimulated interaction between these ligand-specific PR hybrid proteins. However, strong interaction was detected by addition of both agonist and antagonists, indicating the formation of mixed ligand heterodimers and that both PR partners require ligand for dimerization to occur. Based on electrophoretic gel mobility shift assays (EMSAs), these heterodimers appear to have substantially reduced DNA binding activity. Progestin antagonists inhibit agonist activation of PR at concentrations that are too low to be accounted for by a simple competition mechanism for binding to PR. We propose that antiprogestin inactivation of PR in trans by heterodimerization contributes to the biological potency of these compounds. PMID- 9849966 TI - Differential up-regulation of gap junction connexin 26 gene in mammary and uterine tissues: the role of Sp transcription factors. AB - The mRNA and protein expressions of connexin 26 (Cx26) in rat mammary gland and uterus can be up-regulated during pregnancy as well as by the administration of human CG (hCG). In the present study, we found that the time course and magnitude of Cx26 induction by hCG was different in these two tissues. The molecular mechanism underscoring this difference was therefore investigated. We had previously demonstrated that both Sp1 and Sp3 transcription factors play a functional role in Cx26 expression. By the electrophoretic mobility shift assay, nuclear extracts from both virgin mammary gland and uterus were capable of binding to a labeled oligonucleotide probe that contained the proximal GC box and formed three protein-DNA complexes (C1, C2, and C3). In the mammary gland, pregnancy enhanced the intensity of all three complexes, whereas in the uterine tissue there was a decrease in the C2 and C3 complexes and an emergence of a new major component, C4 complex. In the supershift study, the C1 complex could be supershifted only by an antibody against Sp1, whereas C2, C3, and C4 could all be supershifted by an antibody against Sp3, suggesting a potential presence of Sp3 isoforms of various sizes. We therefore conclude that the basal Sp profiles in virgin mammary gland and uterine tissue are similar. However, in response to pregnancy, the changes in Sp profile are tissue specific and may account for the temporal and quantitative differences between these two tissues in Cx26 induction. PMID- 9849967 TI - Differentially expressed messenger RNA isoforms of the human estrogen receptor alpha gene are generated by alternative splicing and promoter usage. AB - The isolation and characterization of several new human estrogen receptor-alpha (hERalpha) mRNAs are described. Together with those previously identified, they give rise to a total of six hERalpha mRNA isoforms (A-F hERalpha mRNAs). Produced from a single hERalpha gene by multiple promoter usage, all these transcripts encode a common protein but differ in their 5'-untranslated region as a consequence of alternative splicing of five upstream exons (1B-1F). RT-PCR and S1 nuclease mapping analysis of these different hERalpha mRNA isoforms revealed a differential pattern of expression of the hERalpha gene in human tissues and cell types. The A hERalpha mRNA is the main isoform detected in mammary glands or in the tumor cell lines derived from this tissue. In endometrium, the predominant forms are the A and C hERalpha mRNA isoforms, whereas the C and F hERalpha mRNA isoforms are the major forms detected in ovary. Finally, high levels of the E hERalpha mRNA isoform are restricted to the liver with an increased expression in females. Taken together, our results demonstrate that the hERalpha gene is a complex genomic unit exhibiting alternative splicing and promoter usage in a tissue-specific manner. PMID- 9849969 TI - On the development of the islets of Langerhans. AB - Studies of pancreatic development have suggested that the islet cells develop through multihormonal stages. Abundant data have confirmed that multihormonal cells are common during pancreatic development. A number of transcription factors and homeotic proteins have also been found to be important to pancreatic and islet cell development. While one of these factors (Isl1) is important for the development of the dorsal pancreatic bud and mesenchyme, another factor (Pdx1) is needed for growth and branching of both pancreatic buds. Studies of the expression patterns of pancreatic hormones and transcription factors and other marker proteins seem at present to be most compatible with the view that early glucagon and glucagon + insulin expressing cells are precursors to the glucagon cells of the islets while mature B cells arise through differentiation from glucagon-negative precursor cells. Recent data also point to possibilities of local paracrine interactions between islet cell types and the parenchymal tissue during development. PMID- 9849968 TI - Granulocyte macrophage-colony stimulating factor reciprocally regulates alphav associated integrins on murine osteoclast precursors. AB - The integrins alphavbeta3 and alphavbeta3 are expressed reciprocally during murine osteoclastogenesis in vitro. Specifically, immature osteoclast precursors, in the form of bone marrow macrophages, contain exclusively alphavbeta5, surface expression of which declines with commitment to the osteoclast phenotype, while levels of alphavbeta3 increase concomitantly. The distinct functional significance of alphavbeta5 is underscored by the integrin's capacity, unlike alphavbeta3, to mediate both attachment and spreading on ligand, of marrow macrophages, suggesting alphavbeta3 negotiates initial recognition, by osteoclast precursors, of bone matrix. Northern analysis demonstrates changes in the two beta-subunits, and not alphav, are responsible for these alterations. Treatment of early precursors with granulocyte-macrophage colony stimulating factor (GM CSF) leads to alterations in beta3 and beta5 mRNA and alphavbeta5 and alphavbeta3, paralleling those occurring during osteoclastogenesis. Nuclear run on and message stability studies demonstrate that while GM-CSF treatment of precursors alters beta5 transcriptionally, the changes in beta3 arise from prolonged mRNA t1/2. Similar to GM-CSF treatment, the rate of beta5 transcription falls during authentic osteoclastogenesis. In contrast to cytokine-induced alphavbeta3, however, that attending osteoclastogenesis reflects accelerated transcription of the beta3-subunit. Thus, while GM-CSF may participate in modulation of alphavbeta5 during osteoclast differentiation, signals other than those derived from the cytokine must regulate expression of alphavbeta3. PMID- 9849970 TI - Morphogenesis and differentiation of the avian endocrine pancreas, with particular reference to experimental studies on the chick embryo. AB - The avian pancreas has three or four lobes and develops from a dorsal and two ventral buds. The cells that will contribute to formation of the dorsal bud are at first located in the mid-dorsal endoderm, those of the ventral buds in the floor of the foregut. The determination of endoderm to form dorsal and ventral bud derivatives occurs before formation of the buds. The highest concentration of endocrine tissue is in the splenic lobe. The lobes contain A and B islets in which glucagon and insulin cells, respectively, predominate. Islets contain somatostatin and pancreatic polypeptide (PP) cells, both of which also occur scattered in the exocrine parenchyma. Pancreatic endocrine cells arise from endoderm: glucagon, insulin, and somatostatin cells differentiate early, PP cells later. To establish culture conditions suitable for avian insulin cells, the epithelial component of dorsal buds of 5-day chick embryos was cultured under various conditions. At the end of 7 days the proportion of insulin cells was determined. In raising the proportion of insulin cells, Matrigel was superior to collagen gel and a serum-free medium (incorporating insulin, transferrin, and selenium) was superior to a serum-containing medium. Modifications to the serum free medium were tested. Raising the level of glucose or of glucose and essential amino acids increased the proportion of insulin cells. This proportion was also increased by replacement of insulin by insulin-like growth factor-I, whereas addition of transforming growth factor beta1 reduced the proportion. Transfer of explants from poor to favourable culture conditions showed that the improved conditions stimulated quiescent insulin progenitor cells to develop. PMID- 9849971 TI - Development of rat pancreatic islet cells in vitro. AB - Islet cell lines were produced by retroviral transduction of SV-40 T antigen to monolayer cultures of neonatal rat islets. One line, RN0-11, showed evolution of phenotypes in vitro. It evolved from a non-hormone secreting cell line to an insulin secreting line. It further developed glucagon producing capability before it lost all hormone producing phenotypes. At passage 8, RN0-11 cells secreted a small amount of insulin 25 ng/10(6) cells/24 hours. They were unresponsive to glucose and secreted 1.7-1.9 ng of insulin/10(6) cells/2 hours under various levels of glucose. At passage 16, they secreted 2,577 ng of insulin/10(6) cells/24 hours and responded to glucose stimulation in static incubation. The insulin secreted by these cells at 0, 2.8, 5.5, 11.1, 16.7, and 27.7 mM of glucose was 6.46 +/- 2.56, 17.74 +/- 2.66, 32.24 +/- 0.58, 30.66 +/- 1.59, 33.55 +/- 4.83, and 20.95 +/- 2.17 ng/10(6) cells/2 hours respectively. The responsiveness to glucose and the ability to secrete insulin diminished as cells were passaged in culture, and by passage 35 no insulin was detectable in medium under any level of glucose tested. Northern blot analyses also showed corresponding changes of insulin expression in these cells at different passages. In addition, glucagon was detectable at passage 14 by immunocytochemistry and at passage 16 by Northern blot analysis. By passage 35, no insulin or glucagon expression was detectable by Northern blot analysis or immunocytochemistry. Immunocytochemical staining of these cells at passage 14 showed insulin-positive and glucagon-positive cells and cells positive for both insulin and glucagon. Presence of insulin and glucagon in the same cells suggests single clonality of the cell line. The evolution of RN0-11 cells in vitro provides an opportunity to study the development of islet cells. PMID- 9849972 TI - Expression of non-classical islet hormone-like peptides during the embryonic development of the pancreas. AB - Understanding of islet embryogenesis may prove to be key in the design of future therapies for diabetes directed at re-initiating islet growth, with the goal to replace and/or replenish the impaired beta-cell mass in the disease. In this context, studies of islet neurohormonal peptides, known to play a role in the local regulation of islet function, and their expression during islet embryogenesis are important. Here we review our studies on the embryonic islet expression of islet amyloid polypeptide (IAPP) and the PP-fold peptides pancreatic polypeptide (PP), peptide YY (PYY) and neuropeptide Y (NPY). IAPP, which is constitutively expressed in beta- and delta-cells in the adult rat, was found to occur in the assumed pluripotent islet progenitor cell, together with PYY, glucagon, and to a lesser extent with insulin. As development proceeds, the insulin/IAPP phenotype is segregated from that of PYY/glucagon; with the formation of islet-like structures, insulin/IAPP-expressing cells primarily occupy their central portions, while PYY/glucagon-expressing cells are found in their periphery. At the time of formation of islet-like structures, expression of NPY is induced in the insulin/IAPP-containing cells. Whereas NPY-expression ceases at birth, PYY is constitutively expressed in non-beta-cells in the mature rat. Expression of PP is induced just prior to birth in a separate population of islet cells, occasionally co-expressed with PYY. Although a clear role for these peptides during embryogenesis has not been identified, they conceivably could play a role in the control of insulin secretion, islet growth and islet blood flow. PMID- 9849973 TI - Development, differentiation, and regeneration potential of the Vervet monkey endocrine pancreas. AB - Using immunolabelling techniques, characterization of the Vervet monkey pancreas included a study of both its development and its structure and normal functioning in the adult. We found that PP and somatostatin occurred during development before glucagon. Insulin and all four pancreatic peptides occurred in one of the primordial buds prior to fusion. These finding call into question the suggested contribution of only PP cells by the ventral bud and non-PP cells by the dorsal bud. Co-localization of glucagon and PP was observed extensively with their relative expressions occurring in what appeared to be an organised non-random manner. Cells expressing both glucagon and PP persisted in the adult, together with many other combinations, suggesting an interesting plasticity of endocrine cell differentiation in the adult. Cellophane wrapping of the head of the Vervet monkey pancreas was shown to result in a noticeable increase in duct cell proliferation and endocrine cell volume but no increased replication of endocrine cells. Cells, immunoreactive for pancreatic peptides, were observed to bud from the ducts, suggesting a regeneration of endocrine cell tissue by neogenesis, although it is uncertain whether the duct epithelium contains the only stem cell source of new endocrine tissue. Hopefully, further investigations will elucidate a mechanism by which endocrine cell regenerative capacity can be stimulated in diabetics to overcome their absolute or relative deficiencies of insulin production. PMID- 9849974 TI - Transdifferentiation versus stem cell hypothesis for the regeneration of islet beta-cells in the pancreas. AB - The pancreas is composed of at least three types of differentiated tissue: the hormone-containing cells in islets (4 different cell types), the exocrine zymogen containing acini, and the centroacinar cells, ductules and ducts (ductal tree). All of these cells appear to have a common origin during embryogenesis in the form of duct-like protodifferentiated cells. Later in life, the acinar and ductal cells retain a significant proliferative capacity that can ensure cell renewal and growth, whereas the islet cells become mitotically inactive. Interestingly, new islet cells, including the insulin-producing beta-cells, can regenerate after tissue injury by a process called neogenesis. The neogenetic process involves differentiation of duct-like (exocrine) epithelial cells to hormone-expressing cells. In this paper, we review the question whether islet beta-cell regeneration or neogenesis in the pancreas depends on "embryonic-like" stem cells or on transdifferentiation of "fully differentiated" cells. This issue is important to find the right model for in vitro research aiming at controlling the process of beta-cell neogenesis. The latter could lead to applications in the treatment of diabetes where functional beta-cells are deficient. We conclude from the available evidence that there is as yet no evidence for the existence of "dormant" stem cells in the adult pancreas. There is some evidence, however, that differentiated exocrine acinar and/or duct cells retain the capacity to transdifferentiate into insulin-expressing beta-cells. PMID- 9849975 TI - Induction of islet cell neogenesis in the adult pancreas: the partial duct obstruction model. AB - The proliferative capacity of adult pancreatic islet cells is limited, although the formation of new islets from cells associated with the ductal epithelium is achievable even in the adult gland. Understanding the mechanism whereby proliferation and subsequent differentiation of putative precursor cells leads the appearance of new islets, i.e., islet neogenesis, may be important as a modality for treatment of both Type I and type II diabetes, in which there is an absolute or relative deficiency of insulin. It appears that certain genes and their protein products are essential to the initiation of the initial step in the pathway. We have shown that partial obstruction of the hamster pancreas is able to reverse streptozotocin-induced diabetes more than 50% of the time. An extract, termed ilotropin, prepared from obstructed pancreata, also reverses the diabetes, whereas extracts of control non-obstructed pancreata do not. Ilotropin contains a protein that is heat and acid stable with MW around 20-45 kDa that is capable of stimulating the proliferation of isolated duct cells in culture. Using mRNA and a differential display technique, 20 genes were found to be expressed in the partially obstructed (regenerating), but not the non-obstructed (non regenerating) pancreas. One of these islet neogenesis-associated proteins (INGAP) proved to be unique to the obstructed pancreas, and a peptide contained within the sequence was capable of stimulating the proliferation of ductal cells in culture. INGAP was found to be expressed early in the neogenic process before the onset of ductal cell proliferation, and was capable of stimulating tritiated thymidine uptake into protodifferentiated epithelial cells, compatible with the notion that it might be involved in initiating the process of islet neogenesis. PMID- 9849976 TI - Foetal rat pancreatic transplantation: posttransplantation development of foetal pancreatic iso- and allografts and suppression of rejection with mycophenolate mofetil (MMF) and cyclosporine based immunesuppression. AB - Provided engraftment can be ensured, vascularization promoted, and ischaemic damage due to storage prevented, foetal pancreatic transplantation (FPT) has the potential to ameliorate the endocrine and metabolic disturbances in diabetic animal models including hyperglycaemia. In a syngeneic Wistar rat substrain (WAG) model (WAG --> WAG), FPT was capable of restoring normoglycaemia in diabetic rats rendered diabetic by streptozotocin (STZ). Posttransplantation growth and development of the foetal tissue was characterised by acinar atrophy, preservation of islet tissue, and development and proliferation of fat accumulations at the site of engraftment. B- and A-cell staining and distribution on ICC appeared normal after 12 months. Mycophenolate mofetil (MMF) together with cyclosporine (CSA) was selected to suppress rejection of foetal rat pancreatic allografts in a strong responder allogeneic model (WAG --> Sprague-Dawley). MMF, a novel immunosuppressive agent that selectively inhibits de novo purine synthesis, was administered in combination with subtherapeutic doses of CSA (2 mg/kg/day) to prevent rejection after allogeneic foetal rat pancreatic transplantation. Although CSA monotherapy in this model can partially suppress rejection, the combination of CSA and MMF results in significant inhibition of acute allograft rejection and mononuclear cellular (MNC) infiltration as assessed by sequential histology post-operatively. Although the follow-up period of allografts was restricted to 30 days of treatment, histology showed low graft infiltrate scores (1.2+) and preservation of islets and immunocytochemical staining. The results in this animal transplantation model confirm that sub therapeutic doses of MMF and CSA therapy are effective in preventing acute rejection of foetal rat pancreatic allografts in the short-term, thus allowing preservation of vital endocrine components of the foetal pancreas such as islets. PMID- 9849977 TI - Alternative algorithms for prevention of perinatal group B streptococcal infections. PMID- 9849978 TI - Safety, tolerability and immunogenicity of concomitant injections in separate locations of M-M-R II, VARIVAX and TETRAMUNE in healthy children vs. concomitant injections of M-M-R II and TETRAMUNE followed six weeks later by VARIVAX. AB - OBJECTIVES AND STUDY DESIGN: The primary objectives of this study were to compare immunologic responses, antibody persistence, safety and varicella breakthrough rates when VARIVAX (varicella vaccine) is given at the same time as M-M-R II (measles, mumps, rubella vaccine) and TETRAMUNE (conjugate Haemophilus influenzae type b, diphtheria, tetanus and whole cell pertussis vaccine) at separate injection sites (Group A) vs. VARIVAX given 6 weeks after M-M-R II and TETRAMUNE (Group B). Six hundred nine healthy children, 12 to 23 months of age, were randomized to one of two treatment (immunization) groups (Group A and Group B). Blood for antibody titers was drawn on the day of immunization, 6 weeks after each injection and 1 year later. Local and systemic adverse reactions were recorded. Exposure and cases of varicella were documented through a 1-year follow up period. RESULTS: Measles, mumps and rubella seroconversion rates and geometric mean titers (GMTs) were similar for both treatment groups. Varicella seroconversion rates were also similar between groups. However, varicella GMTs and percent with a varicella-protective level [> or =5.0 glycoprotein (gp) enzyme linked immunosorbent assay (ELISA) units] did not meet the prespecified criteria for similarity were lower for Group A (GMT 10.5; 82.8% > or =5.0 gp ELISA units) than for Group B (GMT 14.5; 91.2% > or =5.0 gp ELISA units). The GMTs between groups for other antibodies were similar. At the 1-year follow-up antibody titers were comparable in both groups and breakthrough varicella cases appeared generally similar. There were fewer local adverse events (AEs) at the VARIVAX injection sites (9.8% and 2.9%, Group A and B, respectively) than at the TETRAMUNE sites (27.9% and 24.0%). Systemic AEs were not statistically different when M-M-R II was administered alone (8.6%) or concomitantly with VARIVAX (8.9%). When VARIVAX was given alone AEs were 1.8%. The rate of fever > or =102 degrees F after M-M-R II and TETRAMUNE administered together was 10.7% on Days 0 to 3 and 23.7% on Days 7 to 21. When VARIVAX was administered alone, the rate of fever was 5.4% on Days 0 to 3 (P = 0.018) and 10.8% on Days 7 to 21 (P<0.001). CONCLUSION: Because the varicella titers were comparable and varicella breakthrough rates generally similar at 1 year in both groups, we expect that the concomitant administration of VARIVAX with M-M-R II and TETRAMUNE has clinical effectiveness similar to that with VARIVAX 6 weeks after the administration of these other two vaccines. VARIVAX appears to be less reactogenic than M-M-R II and TETRAMUNE. PMID- 9849979 TI - Etiology of childhood pneumonia: serologic results of a prospective, population based study. AB - BACKGROUND: To investigate the etiology of pediatric community-acquired pneumonia, we conducted a prospective, population-based study covering the total population <15 years of age (n = 8851) in 4 municipalities in eastern Finland. MATERIALS AND METHODS: The number of patients was 201; chest radiographs were available for all cases and paired sera for serologic assays were available for >90% of cases. The methods included assays for antibody response to 3 pneumococcal antigens, specific pneumococcal immune complex assays and conventional antibody tests for mycoplasmal, chlamydial and viral infections. RESULTS: Serologic evidence of specific microbial etiology was obtained in 133 (66%) of the pneumonia patients. Bacterial infection was diagnosed in 102 cases (51%) and viral infection in 51 cases (25%). Streptococcus pneumoniae was the most common agent (57 cases; 28%), followed by Mycoplasma pneumoniae (44; 22%), respiratory syncytial virus (43; 21%) and Chlamydia spp. (29; 14%). Haemophilus influenzae was identified in only 6% and Moraxella catarrhalis in only 3% of the children. More than one specific infection was found in 51 patients (25%). The proportion of pneumococcal cases varied from 24 to 36% by age. Mycoplasma infections were seen mostly in patients > or =5 years and Chlamydia infections in patients > or =10 years of age. CONCLUSIONS: The results of our prospective, strictly population-based study confirm the importance of S. pneumoniae in the etiology of community-acquired pneumonia in children of all ages. M. pneumoniae and Chlamydia pneumoniae are important from the age of 5 years onwards. PMID- 9849980 TI - Pneumococcal immune complexes in the diagnosis of lower respiratory infections in children. AB - BACKGROUND: In recent years serologic methods have been applied to assess pneumococcal etiology of pneumonia and other respiratory tract infections. Antigen and antibody assays have shown to be insensitive, especially in young children. The aim of this study was to evaluate the usefulness of circulating immune complexes in the diagnosis of pneumococcal lower respiratory infection in children. MATERIAL AND METHODS: Pneumococcal immune complexes (IC) containing antibodies to species-specific C-polysaccharide, to mixtures of type-specific capsular polysaccharides or to a protein antigen, pneumolysin, were studied in the sera of 449 children with lower respiratory tract infection. RESULTS: Circulating ICs were found in 68 (15%) children; 46 (68%) of them were demonstrated in acute and 43 in convalescent serum. In 5 (7%) of the 68 IC positive patients pneumococcal antigen was present in acute serum; those patients formed 18% of the 28 cases with antigenemia. An antibody response between paired sera to any of the 3 pneumococcal antigens studied was observed in 14 (21%) IC positive children; they formed 23% of the 60 cases with an antibody response. In total ICs were positive in 51% of all the 134 pneumococcal cases diagnosed by any method. CONCLUSIONS: We conclude that the measurement of circulating ICs is more sensitive than other serologic methods for the diagnosis of pneumococcal lower respiratory infection. In infants, however, it was as insensitive as antigen and antibody assays. PMID- 9849981 TI - Epidemiology of respiratory syncytial virus infection requiring hospitalization in East Denmark. AB - BACKGROUND: Prophylaxis against infection caused by respiratory syncytial virus (RSV) with high titered RSV immunoglobulin or humanized antibody may soon be available in Europe. OBJECTIVE: To study the epidemiology of RSV infections requiring hospitalization in infants <6 months in East Denmark to provide a rational basis for decisions concerning prophylaxis against RSV. METHOD: Populat ion-based retrospective review of case records of infants <6 months admitted to pediatric departments with RSV infection in East Denmark from November 1, 1995, to April 30, 1996. RESULTS: Data were obtained from 459 infants. Seventy-three had predisposing conditions: prematurity, 49; pulmonary disease, 2; congenital heart disease, 7; neurologic disease, 6; others, 9. One preterm infant had bronchopulmonary dysplasia. The incidence of RSV infection requiring hospitalization in East Denmark among infants <6 months was estimated to be 34/1000/season. It was 32/1000/season among term infants and 66/ 1000/season among preterm infants (P<0.001). Infants with predisposing conditions and/or nosocomial infection (n = 24) had significantly more severe courses than otherwise healthy infants (P<0.01). One-hundred thirty infants received respiratory support by nasal continuous positive airway pressure, but only six required mechanical ventilation. No infants died. CONCLUSION: The course of RSV disease in East Denmark was milder than reported elsewhere, possibly as a result of the low prevalence of bronchopulmonary dysplasia in Denmark. However, RSV constitutes a considerable burden to the Danish pediatric health care system, and therefore prophylaxis against RSV is desirable. PMID- 9849982 TI - Persistence of antibodies in children after intradermal or intramuscular administration of preexposure primary and booster immunizations with purified Vero cell rabies vaccine. AB - BACKGROUND: The use of intradermal (i.d.) injections of purified Vero cell rabies vaccine (PVRV) for preexposure prophylaxis has not been well-established. We studied the safety and immunogenicity of i.d. and intramuscular (i.m.) PVRV injections for primary and booster preexposure immunizations. METHODS: One of two rabies preexposure PVRV regimens comprising three doses of either 0.1 ml i.d. or 0.5 ml i.m. administered during 28 days was assigned at random to 190 school children. One booster dose was given 1 year later either i.d. or i.m., according to their initial randomization group. Serologic results were available from 155 (82%) children at 1 year after primary immunization and 118 (62%) children at 2 years after booster. RESULTS: Although children vaccinated i.d. had significantly lower rabies-neutralizing antibody titers after primary immunization as well as after booster than children vaccinated i.m. (P< 0.001 for all time points), there were no significant differences in the percentages of children with adequate titers (> or =0.15 IU/ml) between the i.d. and i.m. groups after both primary and booster immunizations. Mild local reactions were more frequent after i.d. vaccination. Mild or moderate systemic reactions were infrequent and similar after i.d. and i.m. vaccinations. Fever and headache were reported by < or =6%. The reactions after booster were not different from those of post-primary immunization. CONCLUSIONS: Purified Vero cell rabies vaccine appears to be safe and immunogenic for primary and booster preexposure immunizations. An i.d. PVRV preexposure regimen should be useful especially for rabies-endemic countries with low per capita income. PMID- 9849984 TI - Candida fungemia in neonates treated with fluconazole: report of forty cases, including eight with meningitis. AB - PURPOSE OF THE STUDY: To assess efficacy and safety of fluconazole in neonates with Candida fungemia. STUDY DESIGN: Multicenter prospective protocol of all fungemias appearing between January 1, 1993, and December 31, 1997, in four major university hospitals. RESULTS: Forty neonates, 28 of them with very low birth weight (<1500 g; 30.5 median gestation week), with documented Candida albicans fungemia were treated with intravenous fluconazole in a daily dosage of 6 mg/kg once daily for 6 to 48 days. Thirty-four received fluconazole as monotherapy and 6 received it in combination with amphotericin B. Thirty-two (80%) were cured; 4 of them relapsed despite at least 14 days of therapy, but they were ultimately cured without sequelae. Eight other neonates died, 4 because of fungal infection and 4 because of prematurity or hemorrhage or lung failure, with fungemia (20% overall and 10% attributable mortality). Two neonates had elevated liver enzymes during fluconazole therapy and 2 others had elevated serum creatinine during fluconazole monotherapy. In none of them did these abnormalities necessitate discontinuation of antifungal therapy. In 8 neonates fungal meningitis developed as a complication of fungemia. All but 3 fungemias were C. albicans; 3 were Candida parapsilosis. CONCLUSIONS: Fluconazole was safe and effective antifungal therapy even in complicated or Candida fungemia in neonates and in infants with very low birth weight. PMID- 9849985 TI - Preoperative diagnosis of Mycobacterium avium lymphadenitis in two immunocompetent children by polymerase chain reaction of gastric aspirates. AB - BACKGROUND: Analysis of gastric aspirates is a routine procedure for detection of Mycobacterium tuberculosis in pediatric pulmonary tuberculosis. However, identification of nontuberculous mycobacteria in gastric aspirates of immunocompetent children is not thought to be clinically significant. METHODS: A PCR method was devised for the detection of M. avium in clinical specimens. The method is based on the amplification of a M. avium-specific DNA fragment present in the 3'-end of the repetitive element IS1245. Surgically removed lymphatic tissue was analyzed prospectively by microscopy, culture and PCR in 13 children admitted to our hospital with suspected mycobacterial lymphadenitis. In 4 of these children 1 to 4 gastric aspirates were obtained before surgical treatment and submitted to the same analysis. RESULTS: We report the detection of M. avium in the gastric aspirates of two children with cervical lymphadenitis before surgical intervention by a novel PCR method. The subsequently surgically removed lymph nodes were also positive by PCR and culture. In one child cultures of both sources grew M. avium. The isolates could be identified as the same strain by DNA fingerprinting. The PCR assay was almost twice as sensitive as culture in detecting M. avium. CONCLUSIONS: Our findings suggest the possibility for noninvasive diagnosis of cervical lymphadenitis caused by nontuberculous mycobacteria before surgery. In addition detection of M. avium in gastric aspirates without evidence of fistula formation provides new insights into the pathogenesis of mycobacterial infection and disease in immunocompetent children. PMID- 9849986 TI - Epidemiology and outcome of osteomyelitis in the era of sequential intravenous oral therapy. AB - BACKGROUND: There has been little recent information on the epidemiology and outcome of childhood osteomyelitis since the era of sequential intravenous-oral antibiotic therapy. METHODS: Retrospective chart review. OBJECTIVE: To review the recent experience and outcome of osteomyelitis in children. RESULTS: Between 1984 and 1996 we identified 146 patients with osteomyelitis. Decreased limb use and fever were the most common presenting symptoms, and tenderness was the most common sign. Staphylococcus aureus was the most common causative organism, and Haemophilus influenzae was not identified after 1990. Bone biopsies or aspirates were superior (83%) to blood cultures (32%) in yielding organisms. 99mTc bone scan was the most sensitive imaging test (94%). Lack of use of anti-teichoic acid titers and serum bactericidal titers did not adversely affect outcome. The median duration of intravenous antibiotic therapy was 11 days, with a median duration of total antibiotics of 38 days. Complications occurred in 6.6% of patients. CONCLUSION: S. aureus remains the most important cause of osteomyelitis. Despite inconsistent use of tests for diagnosis and management and use of sequential intravenous-oral therapy, the overall outcome remains excellent. PMID- 9849983 TI - Management of neonatal candidiasis. Neonatal Candidiasis Study Group. AB - OBJECTIVE: To identify areas of consensus and controversy in the management of neonatal candidiasis. METHODS: A questionnaire was distributed to US-based members of the Pediatric Infectious Diseases Society and a sampling of US neonatologists. RESULTS: Three hundred eighty evaluable questionnaires were returned (42% of those mailed). Ninety-five percent of respondents have cared for an infant with systemic candidiasis in the past 2 years. Fluconazole and liposomal amphotericin are used to some extent by 90 and 69% of respondents, respectively. A single blood culture positive for Candida led to a recommendation for immediate treatment by 99%; amphotericin B was the preferred therapy for candidemia (88%). More than 80% of respondents would request cerebrospinal fluid, urine and repeat blood cultures and ophthalmologic examination in the evaluation of candidemia. If a cerebrospinal fluid culture is positive, 25% would use amphotericin B alone whereas 62% would add flucytosine. For candiduria Society members chose fluconazole therapy more often than did neonatologists, 23% vs. 3.4% (P<0.001). There was no consensus concerning duration of therapy, use of an amphotericin B test dose or management of a central catheter in place during candidemia. CONCLUSIONS: Systemic candidiasis in neonates is a frequently encountered clinical problem. There is agreement that prompt therapy with amphotericin B is required if a blood culture is positive for Candida and that such infants require additional evaluations. Other antifungals (fluconazole, liposomal amphotericin B) are used to some extent in this population. Many issues in management have no clear consensus and warrant further research. PMID- 9849987 TI - Safety and immunogenicity of a combined diphtheria-tetanus-acellular pertussis inactivated polio vaccine-Haemophilus influenzae type b vaccine administered at 2 4-6-13 or 3-5-12 months of age. AB - METHODS: In an open randomized study we compared the safety and immunogenicity of two schedules for priming and booster vaccinations of infants. A pentavalent combination vaccine, including a lyophilized Haemophilus influenzae type b tetanus toxoid conjugate vaccine reconstituted with a liquid diphtheria, tetanus, acellular pertussis (pertussis toxoid and filamentous hemagglutinin) and inactivated polio vaccine (DTaP-IPV/Act-HIB; Pasteur Merieux Connaught, Lyon, France) was administered to 236 Swedish infants either at 2, 4 and 6 months or at 3 and 5 months, and a booster dose was administered 7 months after the last primary dose. Adverse events were monitored by diaries for 3 days after each vaccination and by questions at the ensuing visits. Antibodies against the different vaccine components were analyzed after the primary series of vaccinations, before and after the booster injections. RESULTS: There were no serious adverse reactions, and the rates of febrile events and local reactions were low in both groups. The three dose primary schedule induced higher geometricmean concentrations for all antigens than did the two dose schedule, but there were no differences between the groups in proportions with protective antibody titers against diphtheria, tetanus, Hib and polio or in proportions with certain defined levels of pertussis antibodies. Prebooster results showed a similar pattern, with the exception that the group primed with three injections showed higher proportions of infants with detectable antibodies against polio virus types 1 and 3. After booster vaccinations there were no differences between the two schedules in geometric mean or in proportions with antibodies above defined antibody concentrations, indicating effective priming from both primary series of vaccinations. Conclusion. The combined vaccine DTaP-IPV/ Act-HIB vaccine was equally safe and immunogenic when administered according to both time schedules studied. PMID- 9849989 TI - Low yield of bacterial stool culture in children with nosocomial diarrhea. AB - OBJECTIVE: To determine whether bacterial stool cultures (BSC) are useful in initial evaluation of children with symptoms of nosocomial diarrhea. To answer this question we performed a retrospective record review to determine the yield of BSC in children who developed diarrhea after the third hospital day (HD-3). METHODS: The hospital computer record keeping system was utilized to compile the result of BSC collected from children and adolescents ages 0 to 20 years between January 1, 1988, and October 31, 1996. All specimens were analyzed for Salmonella, Shigella, Yersinia and Campylobacter. We reviewed hospital charts of all children who developed a positive BSC beyond HD-3 to determine the time of onset of diarrhea and clinical circumstances. RESULTS: A total of 11 516 BSCs were submitted from 9262 children during the 8 1/2-year period. Five hundred sixty-eight (6.6%) of 9262 children had at least 1 positive BSC. Two thousand five hundred seventy-two children had the first BSC submitted after HD-3 and 13 (0.5%) of these children had a positive result. Chart review of these 13 children demonstrated that 6 had onset of diarrhea during the first 3 hospital days. Therefore only 7 children met our criteria for having nosocomially acquired diarrhea caused by a bacterial pathogen. Children whose first BSC was submitted after HD-3 accounted for 3767 (46%) of the total 8126 inpatient BSCs and in excess of $21000 annually in patient billing charges. CONCLUSION: In the absence of a known exposure the isolation of a bacterial pathogen from the stool of children with onset of diarrhea beyond HD-3 is a rare event. Under most circumstances BSC should not be part of the initial evaluation of children with symptoms of nosocomial diarrhea. PMID- 9849988 TI - Measles vaccine effectiveness in standard and early immunization strategies, Niger, 1995. AB - BACKGROUND: An Expanded Programme on Immunization was started in late 1987 in Niger, including vaccination against measles with one dose of standard titer Schwarz vaccine given to infants after 9 months of age. During epidemics an early two-dose strategy was implemented (one dose between 6 and 8 months and one dose after 9 months). From January 1, 1995, until May 7, 1995, 13 892 measles cases were reported in Niamey, Niger. METHODS: A retrospective cohort study was conducted in a crowded area of Niamey at the end of the outbreak to assess the effectiveness of measles vaccine in standard (after 9 months) and early (before 9 months) immunization strategies under field conditions. RESULTS: Highest measles incidence rates were observed among children <1 year of age. Vaccine effectiveness estimates increased with age at vaccination from 78% with a single dose administered at 6 months of age to 95% at 9 months. Vaccine effectiveness with the early two dose strategy was 93%. CONCLUSIONS: Immunization with a single dose of standard titer Schwarz vaccine before 9 months of age provided higher clinical protection than expected from seropositivity studies. The early two dose strategy is justified in contexts where measles incidence is high before 9 months of age. Our results raise the issue of lowering the recommended age for measles vaccination in developing countries. PMID- 9849990 TI - Hepatitis G Virus. PMID- 9849991 TI - Chlamydia pneumoniae revisited. PMID- 9849992 TI - Antibiotic usage in appendicitis in children. PMID- 9849993 TI - Treatment of neonatal chlamydial conjunctivitis with azithromycin. PMID- 9849994 TI - Chromosomal proviral sequences of human immunodeficiency virus in multiple thymic cell types in a patient with acquired immunodeficiency syndrome and massive thymic hyperplasia. PMID- 9849995 TI - Pyomyositis and septic arthritis caused by group C Streptococcus. PMID- 9849996 TI - Acute aseptic meningitis secondary to intravenous immunoglobulin in a patient with Kawasaki syndrome. PMID- 9849997 TI - Nosocomial myiasis in an extremely premature infant caused by the sheep blowfly Lucilia sericata. PMID- 9849998 TI - An adolescent with nephrotic syndrome and multiple leg nodules. PMID- 9849999 TI - Current recommendations on the therapy of otitis media. PMID- 9850000 TI - Therapeutic options for cat-scratch disease. PMID- 9850001 TI - Pharmacokinetics and pharmacodynamics of new and old antimicrobial agents for acute otitis media. AB - Selection of appropriate antibiotic treatment for children with acute otitis media (AOM) is challenging. Although the diagnosis is relatively easy for experienced clinicians, the distinction between AOM and otitis media with effusion is often more subtle. In general therapy is empiric and the pathogen causing disease in a given patient remains unknown. However, this situation is made even more difficult by the dynamic nature of the pathogenesis of AOM. Both the proportion of patients infected with one of the three primary pathogens, Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, and the antimicrobial susceptibility patterns of these pathogens are changing. Currently there are 16 antibiotics labeled for use in AOM. Only 2 are reliably effective against penicillin-resistant pneumococcus: high dose amoxicillin (80 to 100 mg/kg/day) and im ceftriaxone. Among the others all are beta-lactamase-stable and have proven clinical effectiveness in AOM patients infected with H. influenzae or M. catarrhalis. Even with the high spontaneous resolution rate reported for AOM, antimicrobial therapy remains the standard of care in the United States. Recognition of the fundamental determinants of effective therapy should permit rational antibiotic selection for each patient. PMID- 9850002 TI - Otitis media and child development: should we worry? AB - Persistent middle ear effusion in infancy and early childhood has been blamed for impairments of speech, language, cognition and psychosocial development later in life. Whether that blame is justified remains unresolved and a matter of controversy, because studies of the relationships involved have been limited and often contradictory and because none was designed so as to address the issue of causality. At issue in particular is the common practice of subjecting infants and young children with persistent otitis media with effusion (OME) to tympanostomy tube placement specifically to reduce the risk of developmental impairment. Currently children younger than age 3 years undergo an estimated 313000 tympanostomy tube placement operations per year, at a cost of about $750 million. If a causal association between early life OME and later developmental impairment were to be established, answers would also be needed to the questions whether the adverse effects of OME are linear or threshold, whether they are permanent or transient and whether they are preventable by timely tube placement. A prospective study designed to address all of these questions is currently under way at Children's Hospital of Pittsburgh. The study involves enrolling a large, demographically diverse sample of normal infants before 2 months of age; monitoring them for the presence or absence of otitis media throughout the first 3 years of life; identifying those in whom OME has persisted for specified minimum periods; randomly assigning those subjects either to prompt tube placement or to delayed tube placement if OME persists; and administering a battery of standardized developmental tests to those subjects and to a sample of the others at ages 3, 4 and 6 years. Details of the study design and procedures are described in this report. PMID- 9850003 TI - Clinical implications of antibiotic resistance for management of acute otitis media. AB - Antibiotic resistance to available antimicrobial agents has been constant since the introduction of the sulfonamides in the 1930s. Multidrug-resistant Streptococcus pneumoniae and beta-lactamase-producing Haemophilus influenzae are a concern now because of the importance of these pathogens in infections of the respiratory tract in infants and children. Amoxicillin remains the drug of choice for initial episodes of acute otitis media (AOM) although increase of the dosage schedule to 80 mg/kg/day has been recommended by some investigators. There are 15 additional antimicrobial agents approved by the Food and Drug Administration for the indication of AOM. All approved drugs are clinically effective but some have been suggested to have priority for patients who fail amoxicillin: amoxicillin clavulanate; an oral cephalosporin such as cefuroxime axetil; and intramuscular ceftriaxone. Management of the child with severe and recurrent disease should include antibiotic prophylaxis but the increased incidence of resistance requires selective use. Prevention of infection may be achieved by innovative techniques for interference with attachment of bacteria to the nasal mucosa such as administration of oligosaccharides in a nasal spray. The currently available polysaccharide pneumococcal vaccines have limited immunogenicity in infants, but the vaccine is useful in children 2 years of age and older who still have recurrent AOM. Children with frequent AOM during the prior respiratory season are candidates also for influenza virus vaccine. If medical management fails to prevent new episodes of AOM in children with severe and recurrent disease, placement of tympanostomy tubes and possible adenoidectomy should be considered. PMID- 9850004 TI - Role of surgery for otitis media in the era of resistant bacteria. AB - Otitis media is currently the most common diagnosis made by clinicians, and its prevalence has an impact on managed health care. With the emergence of bacterial pathogens resistant to many antimicrobial agents, an urgent need exists to reassess the indications for surgical management of this more prevalent disease. In an effort to determine the causative bacterial etiology of acute otitis media, which may be resistant to commonly prescribed antimicrobial agents, tympanocentesis is indicated today more than ever, especially when patients fail to improve or worsen while receiving antibiotics (i.e. treatment failures). In an effort to reduce the use of antimicrobial agents, prophylactic administration of these drugs for prevention of recurrent otitis media should only be prescribed on an individualized basis, with myringotomy and tympanostomy tube insertion being a more reasonable alternative. Adenoidectomy should be also considered when moderate to severe nasal obstruction is present or when repeat tympanostomy tube placement is needed. Likewise when chronic otitis media with effusion is unresponsive to a trial of antimicrobial therapy, tympanostomy tube insertion, adenoidectomy or both procedures should be considered, as opposed to re-treating with a variety of antibiotics. The role of these surgical procedures has become more important today in this era of antibiotic-resistant bacteria. Also in our cost-conscious environment, it is uncertain which method of management is more expensive, surgery or prolonged medical management. PMID- 9850006 TI - Spermidine determines the sensitivity to the calmodulin antagonist, chlorpromazine, for the circadian conidiation rhythm but not for the mycelial growth in Neurospora crassa. AB - The gene that suppresses the phenotype of the cpz-2 mutation, which results in changing the sensitivity to chlorpromazine in relation to mycelial growth and circadian rhythms, was cloned in Neurospora crassa. This gene is not the cpz-2 gene itself but rather is identical to the spe-3 gene that encodes spermidine synthase in Neurospora. The intracellular content of spermidine was lowered in the cpz-2 strain compared to that of the wild-type strain. By integration of the spe-3 gene or by the addition of spermidine into culture medium, the temperature sensitivity of mycelial growth was lost and the conidiation rhythm became sensitive to chlorpromazine in the cpz-2 strain, as was observed in the wild-type strain, but the hypersensitivity of mycelial growth on chlorpromazine in the cpz 2 strain was not affected. Therefore, it appears that spermidine determines only the sensitivity of the conidiation rhythm to chlorpromazine. PMID- 9850007 TI - Thermoperiodic responses in insects and mites simulated with the double circadian oscillator clock. AB - The "double circadian oscillator model" for the photoperiodic clock has been used to simulate thermoperiodic responses in insects and mites. Two assumptions have been made: (1) the clock measures cryophase in a similar way to scotophase, and (2) temperature cycles are able to entrain the clock in a similar way to LD cycles. Simulations showed that Assumption 1 causes the "critical cryophase" to be of about equal duration as the critical night length. Assumption 2 is not always needed if diapause incidences in DD are high at low temperatures but low or zero at high temperatures. The latter assumption is needed, however, if high diapause occurs in thermoperiodic cycles in DD, whereas nondiapause occurs in DD with both high and low constant temperatures. The model accounts for the observation that the amplitude of the temperature cycle is important in some insects, whereas the temperature of the cryophase is crucial in others. PMID- 9850008 TI - Effects of circadian mutations and LD periodicity on the life span of Drosophila melanogaster. AB - The pervasive occurrence of circadian clocks throughout the living world underlines their adaptive value. Nonetheless, there is surprisingly little evidence for a negative impact, on any animal species, of a constant discrepancy between the environmental and endogenous periods. Male Drosophila melanogaster per mutants with altered circadian periods were compared to the wild type in two different LD schedules. Life span was used as a global index of physiological adaptation. The life span of the mutants was significantly reduced by up to 15% for the flies whose period differs most from that of the wild type. A reduction was observed even when flies were kept in an LD schedule fitting a mutant period. The LD schedule made no significant difference on its own, but the authors found evidence for an interaction between genotype and LD schedule in determining life span. These results are consistent with the importance of the circadian clock in maintaining internal temporal order independent of environmental cycles. Nonetheless, a large difference between the environmental and endogenous periods has a measurable impact. PMID- 9850009 TI - Day/night differences in the stimulation of adenylate cyclase activity by calcium/calmodulin in chick pineal cell cultures: evidence for circadian regulation of cyclic AMP. AB - In chick pineal cell culture, stimulation of adenylate cyclase with the diterpene forskolin was greater during the subjective night than during the subjective day. This rhythm of cyclic AMP (cAMP) stimulation mimicked the rhythm of unstimulated cAMP measured previously during LD cycles from flow-through culture. Direct measurement of adenylate cyclase activity in permeabilized cells revealed an adenylate cyclase activity activated by Ca2+/calmodulin during the night but not during the day. However, this difference in adenylate cyclase activity at two times of the circadian cycle is apparent only when permeabilized cells were prewashed with buffer containing GTE When cAMP was measured from flow-through cultures maintained in continuous darkness to determine whether a circadian clock may regulate cAMP, a low-amplitude rhythm was measured. The circadian rhythm of cAMP was similar to the cAMP rhythm previously measured on LD cycles except that the rhythm in darkness had a lower amplitude. Similar to the suppression of melatonin, cAMP was suppressed by light presented during the middle of the night. LD differences in nocturnal cAMP levels were abolished with dipyridamole, an inhibitor of cyclic GMP (cGMP) phosphodiesterase. These results suggest that the rhythm of cAMP in chick pineal cells involves the stimulation of adenylate cyclase by Ca2+/calmodulin during the night and a GTP-dependent suppression of adenylate cyclase activity during the day. The photic suppression of cAMP at night involves the activation of a dipyridamole-sensitive, cGMP phosphodiesterase. PMID- 9850010 TI - Destruction of serotonergic neurons in the median raphe nucleus blocks circadian rhythm phase shifts to triazolam but not to novel wheel access. AB - Systematic treatment of hamsters with triazolam (TRZ) or novel wheel (NW) access will yield PRCs similar to those for neuropeptide Y. Both TRZ and NW access require an intact intergeniculate leaflet (IGL) to modulate circadian rhythm phase. It is commonly suggested that both stimulus types influence rhythm phase response via a mechanism associated with drug-induced or wheel access-associated locomotion. Furthermore, there have been suggestions that one or both of these stimulus conditions require an intact serotonergic system for modulation of rhythm phase. The present study investigated these issues by making serotonin neuron-specific neurotoxic lesions of the median or dorsal raphe nuclei and evaluating phase response of the hamster circadian locomotor rhythm to TRZ treatment or NW access. The expected effect of TRZ injected at CT 6 h on the average phase advance was virtually eliminated by destruction of serotonin neurons in the median, but not the dorsal, raphe nucleus. No control or lesioned animal engaged in substantial wheel running in response to TRZ. By contrast, all median raphe-lesioned hamsters that engaged in substantial amounts of running when given access to a NW had phase shifts comparable to control or dorsal raphe lesioned animals. The results demonstrate that serotonergic neurons in the median raphe nucleus contribute to the regulation of rhythm phase response to TRZ and that it is unlikely that these neurons are necessary for phase response to NW access. The data further suggest the presence of separate pathways mediating phase response to the two stimulus conditions. These pathways converge on the IGL, a nucleus afferent to the circadian clock, that is necessary for the expression of phase response to each stimulus type. PMID- 9850011 TI - Phase shift magnitude and direction determine whether Siberian hamsters reentrain to the photocycle. AB - Body temperature (Tb) or activity rhythms were monitored in male Siberian hamsters (Phodopus sungorus) housed in an LD cycle of 16 h light/day from birth. At 3 months of age, rhythms were monitored for 14 days, and then the LD cycle was phase delayed by 1, 3, or 5 h or phase advanced by 5 h in four separate groups of animals. Phase delays were accomplished via a 1- or 3-h extension of the light phase or via a 5-h extension of the dark phase. The phase advance was accomplished via a 5-h shortening of the light phase. After 2 to 3 weeks, hamsters that were phase delayed by 1 or 3 h were then phase advanced by 1 or 3 h, respectively, via a shortening of the light phase. All of the animals reentrained to phase delays of 1 or 3 h and to a 1-h phase advance; 79% reentrained to a 3-h phase advance. In contrast, only 13% of the animals reentrained to the 5-h phase advance, 13% became arrhythmic, and 74% free ran for several weeks. After the 5-h phase delay, however, reentrainment was observed in 50% of the animals although half of them required more than 21 days to reentrain. The response to a phase shift could not be predicted by any parameter of circadian rhythm organization assessed prior to the phase shift. These data demonstrate that a phase shift of the LD cycle can permanently disrupt entrainment mechanisms and eliminate circadian Tb and activity rhythms. Magnitude and direction of a phase shift of the LD cycle determine not only the rate but also the probability of reentrainment. Furthermore, the phase of the LD cycle at which the phase shift is made has a marked effect on the proportion of animals that reentrain. Light exposure during mid-subjective night combined with daily light exposure during the active phase may explain these phenomena. PMID- 9850013 TI - Effect of sustained nocturnal transbuccal melatonin administration on sleep and temperature in elderly insomniacs. AB - Previous research has suggested a role for the pineal hormone melatonin in the control of the body's sleep-wake and thermoregulatory systems. In the elderly population, there have been reports of decreased nighttime secretion of melatonin and suggestions that this may, in turn, be responsible for the increased incidence of sleep disorders reported by this age group. On this basis, it has been suggested that augmented nocturnal melatonin levels may improve sleep quality in age-related sleep disorders. Following screening assessments, 12 elderly (> 55 years) subjects with sleep maintenance insomnia were treated with either 0.5 mg transbuccal melatonin or a placebo for two sessions of 4 consecutive nights, at least 3 days apart. Subjects self-selected lights-out times, and sleep was assessed using standard polysomnographic (PSG) measures. Body temperature was measured continually from 2100 to 0700 h, and sleep quality was assessed from PSG variables measured. Nightly urine samples were assayed for the melatonin metabolite 6-sulfatoxy-melatonin (aMT.6S). Compared to the placebo, transbuccal melatonin administration significantly increased mean nocturnal aMT.6S excretion (mean +/- SEM: 194.2 +/- 16.5 vs. 42.5 +/- 7.7 nmol). In addition, there was a significant reduction in core body temperature relative to the placebo condition (p < .05). However, sustained transbuccal melatonin treatment had no positive significant effect on any PSG measure of sleep quality. The results from the present study suggest that sustained nocturnal administration of melatonin, in the low pharmacological range, might be of limited clinical benefit in this subject population. PMID- 9850012 TI - Melatonin chimeras alter reproductive development and photorefractoriness in Siberian hamsters. AB - Nightly melatonin (MEL) durations > 8 h provoke gonadal regression and decreases in body mass, whereas signals < 7 h stimulate gonadal and somatic growth in male Siberian hamsters. The authors sought to determine the minimum frequency of short MEL signals sufficient to induce the long-day phenotype in several photoperiodic traits. D,L-propranolol (hereafter propranolol) injections shortened MEL signals on the night of treatment without altering MEL on the subsequent night; this permitted interpolation of short MEL signals at variable frequencies against a background of long MEL signals (chimeras). Hamsters kept in short days (10 h light/day, 10L) were injected with propranolol 6 h after dark onset for 28 consecutive weeks beginning at 30 days of age (Week 0) either every other day or once every 3, 6, or 9 days. Control animals were injected with saline or with propranolol during the light phase or were transferred to long days (16L) at Week 0. Hamsters in 16L underwent rapid gonadal development and increases in body mass and displayed summer pelage color, as did hamsters treated with propranolol every other day. Animals treated with propranolol less frequently than every other day uniformly maintained undeveloped gonads and winter-like body weights, but pelage color became proportionately darker with increased frequency of propranolol treatments. The onset of spontaneous testicular development in 10L was unaffected by propranolol injections. After termination of injections at Week 28, testicular regression was not observed in most 10L animals that previously had undergone spontaneous testicular development; however, 40% of hamsters that had been injected with propranolol every 3rd night did manifest the winter phenotype after Week 28. In an alternating sequence, short MEL signals completely override long signals and induce the summer phenotype. Threshold frequencies differ for MEL stimulation of long-day pelage and gonadal phenotypes. The timing and development of refractoriness to MEL does not depend in any simple manner on the number of long MEL signals or on the accumulation of a reaction product produced by long, and depleted by short, MEL signals. PMID- 9850014 TI - Combination chemotherapy versus single-agent therapy as first- and second-line treatment in metastatic breast cancer: a prospective randomized trial. AB - PURPOSE: We report results of a randomized prospective study that compared single agents of low toxicity given both as the first-line and second-line chemotherapy with combination chemotherapy in advanced breast cancer with distant metastases. PATIENTS AND METHODS: Patients in the single-agent arm (n = 153) received weekly epirubicin (E) 20 mg/m2 until progression or until the cumulative dose of 1,000 mg/m2, followed by mitomycin (M) 8 mg/m2 every 4 weeks, and those in the combination chemotherapy arm (n = 150) were first given cyclophosphamide 500 mg/m2, E 60 mg/m2, and fluorouracil 500 mg/m2 three times per week (CEF) followed by M 8 mg/m2 plus vinblastine (V) 6 mg/m2 every 4 weeks. Exclusion criteria included age greater than 70 years, World Health Organization (WHO) performance status greater than 2, prior chemotherapy for metastatic disease, and presence of liver metastases in patients younger than 50. RESULTS: An objective response (complete [CR] or partial [PR]) was obtained in 55%, 48%, 16%, and 7% of patients treated with CEF, E, M, and MV, respectively. A response to CEF tended to last longer than a response to E (median, 12 v 10.5 months; P = .07). Treatment related toxicity was less in the single-agent arm and quality-of-life (QOL) analysis favored the single-agent arm. No significant difference in time to progression or survival was found between the two arms. Similarly, no difference in survival was found when the patients who received both the planned first-and second-line treatments were compared or when survival was calculated from the beginning of the second-line therapy. CONCLUSION: Patients treated with single agent E followed by single-agent M had similar survival, but less treatment related toxicity and better QOL as compared with those treated with CEF followed by MV. PMID- 9850015 TI - Inflammatory breast cancer and body mass index. AB - PURPOSE: No studies have investigated the etiology of inflammatory breast cancer (IBC), the most lethal form of breast cancer. Because high body mass index (BMI) is associated with decreased risk of premenopausal breast cancer but increased risk of postmenopausal breast cancer, we evaluated whether high BMI was a risk factor for IBC. PATIENTS AND METHODS: In a case-comparison study, we matched by ethnicity and registration date 68 IBC patients treated at The University of Texas M.D. Anderson Cancer Center from 1985 to 1996 with 143 patients with non IBC and 134 patients with cancer at sites other than the breast or reproductive tract (non-breast cancer). The non-breast cancer group was used in lieu of a population-based, healthy control group, which was not available. RESULTS: IBC patients were younger at menarche and the time of their first live birth than non IBC and non-breast cancer patients. The proportion of premenopausal IBC patients was higher than the proportion of premenopausal women in the comparison groups, although differences were not significant. There were no differences in height, but IBC patients were heavier (77.6 kg) than non-IBC (70.0 kg) and non-breast cancer patients (68.0 kg). After adjusting for other factors, women in the highest BMI tertile (BMI > 26.65 kg/m2) relative to the lowest tertile (BMI < 22.27) had significantly increased IBC risk (IBC v non-IBC, odds ratio [OR] = 2.45, 95% confidence interval [CI] = 1.05 to 5.73; IBC v non-breast cancer, OR = 4.52, 95% CI = 1.85 to 11.04). This association was not significantly modified by menopausal status and was independent of age at menarche, family history of breast cancer, gravidity, smoking status, and alcohol use. CONCLUSION: Our investigation showed that high BMI was significantly associated with an increased risk of IBC. This association did not vary by menopausal status, although IBC patients were more likely to be premenopausal. Confirming our findings and identifying other IBC risk factors may provide directions for future research on the aggressive nature of IBC. PMID- 9850016 TI - Ewing's family of tumors in adults: multivariate analysis of survival and long term results of multimodality therapy in 182 patients. AB - PURPOSE: To assess the outcome and the prognosis of adults with a neoplasm related to the Ewing's sarcoma family of tumors. PATIENTS AND METHODS: The outcomes of 182 consecutive patients older than 15 years with Ewing's sarcoma or related neoplasms managed from 1982 to 1992 were reviewed, without any selection according to primary tumor site or disease extension. RESULTS: Of 182 patients, 53 had evidence of metastases at presentation (29%). Tumor size was greater than 10 cm in 70 patients (41%). With a median follow-up duration of 66 months, the 5 year overall survival (OS) rate was 41%. In patients with localized disease, 5 year OS rate was 54% and 5-year progression-free survival (PFS) rate, 43%. Late relapses after 5 years accounted for 9% of relapses. Metastasis at presentation (P = .00001), pelvic primary lesion (P = .0025), and tumor size greater than 10 cm (P = .004) were independent prognostic factors for survival. Five-year OS was 67% in patients with nonpelvic tumors < or = 10 cm, 52% in those with pelvic tumors less than 10 cm or extrapelvic tumors > or = 10 cm, 16% in those with pelvic tumors greater than 10 cm, and 9% in those with metastasis (P = .00001). CONCLUSION: Based on our experience and a review of the literature, we concluded that the natural history and the prognosis of the Ewing's family of tumors in adults are not different from that found in children. A greater tumor bulk in adults may explain the less favorable prognosis previously reported by others. Outcome could be adequately monitored by a simple prognostic index. PMID- 9850017 TI - Effect of duration of treatment on treatment outcome and cost of treatment for Wilms' tumor: a report from the National Wilms' Tumor Study Group. AB - PURPOSE: National Wilms' Tumor Study (NWTS)-4 was designed to evaluate the efficacy, toxicity, and cost of the administration of different regimens for the treatment of Wilms' tumor (WT). PATIENTS AND METHODS: Between August 6, 1986 and September 1, 1994, 905 previously untreated children aged younger than 16 years with stage II favorable histology (FH) WT (low-risk [LR]), stages III to IV FH WT, or stages I to IV clear-cell sarcoma of the kidney (high-risk[HR]) were randomized after the completion of 6 months of chemotherapy to discontinue (short) or continue for 9 additional months (long) treatment with chemotherapy regimens that included vincristine and either divided-dose (standard [STD]) courses (5 days) or single-dose (pulse-intensive [PI]) treatment with dactinomycin. HR patients also received either divided-dose (STD) courses (3 days) or single-dose (PI) treatment with doxorubicin. RESULTS: The 4-year relapse free survival (RFS) rates after the second randomization for LR patients were 83.7% for the 190 patients treated with short and 88.2% for the 187 patients treated with long chemotherapy (P = .11). The 4-year RFS rates after the second randomization for HR FH patients were 89.7% for the 256 patients treated with short and 88.8% for the 246 patients treated with long chemotherapy (P = .87). The charge for treatment with the short PI treatment regimens for all children with stages I through IV FH WT was approximately one half of that with the long STD treatment regimens. CONCLUSION: The short administration schedule for the treatment of children with WT is no less effective than the long administration schedule and can be administered at a substantially lower total treatment cost. PMID- 9850018 TI - Bone mass after treatment for acute lymphoblastic leukemia in childhood. AB - PURPOSE: To study bone mass after childhood acute lymphoblastic leukemia (ALL) and determine if reduced bone mass is related to previous therapy or endocrine status at follow-up. PATIENTS AND METHODS: We studied 95 survivors of childhood ALL who were in first remission a median of 11 years (range, 3 to 23 years) after diagnosis and who had never been irradiated outside a cranial field. The bone mass was measured by dual-energy x-ray absorptiometry. The results were compared with data on 396 local controls. RESULTS: Adjusted for sex and age, the mean whole-body bone mineral content (BMC) and bone mineral areal density (BMDA) were both significantly reduced (0.4 SDs less than the predicted mean value). This was mainly caused by reduced bone mass in the 33 participants who were aged 19 years or older at follow-up. In these young adults, the mean height for age, bone area for height, and BMC for bone area were all significantly reduced. This indicated that the reduced whole-body bone mass was caused by both reduced bone size and reduced size-adjusted bone mass. Reduced bone size was related to previous cranial irradiation. Reduced size-adjusted bone mass was not significantly related to age at diagnosis or at follow-up, length of follow-up, cranial irradiation, cumulative dose of methotrexate or corticosteroids, or endocrine status at follow-up. CONCLUSION: The whole-body bone mass was reduced 11 years after diagnosis of childhood ALL. If these abnormalities remain, survivors of childhood ALL will have an increased risk for osteoporotic fractures later in life. PMID- 9850019 TI - Secondary brain tumors in children treated for acute lymphoblastic leukemia at St Jude Children's Research Hospital. AB - PURPOSE: To evaluate the incidence of and potential risk factors for second malignant neoplasms of the brain following treatment for childhood acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: The study population consisted of 1,612 consecutively enrolled protocol patients treated on sequential institutional protocols for newly diagnosed ALL at St Jude Children's Research Hospital (SJCRH) between 1967 and 1988. The median follow-up duration is 15.9 years (range, 5.5 to 29.9 y). RESULTS: The cumulative incidence of brain tumors at 20 years is 1.39% (95% confidence interval [CI], 0.63% to 2.15%). Twenty-two brain tumors (10 high-grade gliomas, one low-grade glioma, and 11 meningiomas) were diagnosed among 21 patients after a median latency of 12.6 years (high-grade gliomas, 9.1 years; meningiomas, 19 years). Tumor type was linked to outcome, with patients who developed high-grade tumors doing poorly and those who developed low-grade tumors doing well. Risk factors for developing any secondary brain tumor included the presence of CNS leukemia at diagnosis, treatment on Total X therapy, and the use of cranial irradiation, which was dose-dependent. Age less than 6 years was associated with an increased risk of developing a high grade glioma. CONCLUSION: This single-institution study, with a high rate of long term data capture, demonstrated that brain tumors are a rare, late complication of therapy for ALL. We report many more low-grade tumors than others probably because of exhaustive long-term follow-up evaluation. The importance of limiting cranial radiation is underscored by the dose-dependent tumorigenic effect of radiation therapy seen in this study. PMID- 9850021 TI - Prognostic value of immunophenotypic detection of minimal residual disease in acute lymphoblastic leukemia. AB - PURPOSE: The identification of immunophenotypic aberrancies through multiparametric flow cytometry makes the differentiation between normal and leukemic cells relatively simple and quick, and is therefore an attractive method for the investigation of minimal residual disease (MRD). In this report, we have analyzed the impact on relapse and relapse-free survival (RFS) of detecting immunophenotypical aberrant cells in acute lymphoblastic leukemia (ALL) patients in cytomorphologic complete remission (CR). MATERIALS AND METHODS: Two hundred eleven bone marrow (BM) samples from 53 consecutive ALL (37 precursor B-ALL and 16 T-ALL) patients were analyzed. The only selection criteria were to have at least one aberrant immunophenotypic feature at diagosis and to have achieved cytomorphologic CR after induction therapy. For MRD detection, all follow-up samples were analyzed with triple labelings using a two-step acquisition procedure, in which 106 cells were screened for the possible persistence of residual leukemic cells with the same phenotypic aberrancy as that identified diagnosis. RESULTS: Patients who displayed a gradual increase in MRD levels showed a higher relapse rate (90% v22%; P < .00001) and shorter median RFS (12 months v not reached; P < .0001) than those with stable or decreasing MRD levels. This adverse prognostic influence also was observed when children and adults, as well as B-ALL and T-ALL patients, were analyzed separately. An MRD level > or = or greater than 10(-3) discriminated two risk groups of ALL patients with significantly different relapse rates and RFS at all treatment phases (end of induction, consolidation, maintenance, and out of treatment). CONCLUSION: Multiparametric flow cytometry of MRD in ALL patients is a valuable tool for relapse prediction and for the identification of a cohort of patients with very poor prognosis. PMID- 9850020 TI - Reappraisal of the clinical and biologic significance of myeloid-associated antigen expression in childhood acute lymphoblastic leukemia. AB - PURPOSE: To reassess the clinical and biologic significance of myeloid-associated antigen expression in childhood acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: We prospectively studied 334 newly diagnosed cases of this disease, using a comprehensive panel of antibodies that represented five myeloid cluster groups (CD13, CD14, CD15, CD33, and CD65). Blast cells were tested for ETV6 and MLL rearrangement using Southern blot analysis. RESULTS: CD13 was expressed in 13.7% of cases, CD14 in 1%, CD15 in 6.6%, CD33 in 16%, and CD65 in 9.7%. Approximately one third of cases (31.4%) expressed one or more of these antigens (B-cell precursor, 31.9%; T-cell, 28.8%), while 10.5% expressed two or more (B cell precursor, 11.3%; T-cell, 6.1%). Among the B-cell precursor leukemias, myeloid-associated antigen expression was significantly associated with a lack of hyperdiploidy and rearrangements of ETV6 or MLL gene. Most of the cases with MLL rearrangements (82%) expressed CD65, CD15, and CD33, either alone or in combination, whereas 48% of those with a rearranged ETV6 gene expressed CD13, CD33, or both. Myeloid-associated antigen expression did not correlate with event free survival, whether the analysis was based on any of the five antigens in our panel or on the three more commonly tested antigens (CD13, CD33, and CD65). Importantly, this finding was not affected by exclusion of patients with ETV6 or MLL gene rearrangements. CONCLUSION: Even though blast cell expression of myeloid associated antigen expression shows significant associations with specific genetic abnormalities, it lacks prognostic value in childhood ALL. PMID- 9850022 TI - Determinants of prognosis in late chronic-phase chronic myelogenous leukemia. AB - PURPOSE: Since interferon alfa (IFN-A) became an established treatment in chronic myelogenous leukemia (CML), more patients are referred to tertiary centers in late chronic phase (ie, > 12 months after diagnosis). Trials conducted in this phase cannot be evaluated precisely unless the features that determine prognosis in late chronic-phase CML are identified. The purpose of this study is to define the prognostic determinants of late chronic-phase CML. PATIENTS AND METHODS: From 1980 to 1997,257 consecutive CML patients referred in late chronic phase were studied. Their clinical characteristics at the time of referral and their association with survival were investigated. A staging model was designed. RESULTS: The median survival from time of referral was 43 months. Pretreatment characteristics associated with worse outcome included older age, poor performance status, splenomegaly, low albumin level, high percentage of blasts or basophils in peripheral blood (PB) or bone marrow, longer duration of chronic phase, and poor-risk group as defined by the Synthesis model. Prior exposure to IFN-A was not associated with worse outcome. By multivariate analysis, characteristics associated with shorter survival were age of 60 years or older, time from diagnosis of 3 years or greater, performance status of 1 or greater, PB basophils of 7% or greater, spleen 10 cm or greater, PB blasts 3% or greater, and albumin level less than 4 g/dL. A model that included age, duration of chronic phase, performance status, and PB basophils was generated. Patients with no, one, two, or three or greater adverse factors had median survivals of 71, 49, 26, and 19 months, respectively. CONCLUSION: A staging model for late chronic-phase CML can stratify patients in four groups with significantly different outcomes. If confirmed in independent populations, such a model could be considered in the analysis of future trials of treatment strategies in late chronic-phase CML. PMID- 9850023 TI - Burkitt-like lymphomas in AIDS patients: characterization within a series of 103 human immunodeficiency virus-associated non-Hodgkin's lymphomas. Burkitt's Lymphoma Study Group. AB - PURPOSE: Burkitt-like lymphoma (BLL) is a tumor with morphologic features intermediate between Burkitt's lymphoma (BL) and large-cell lymphoma, but its relationship with these lymphomas is currently unclear. We have therefore analyzed its characteristics within a large series of human immunodeficiency virus (HIV)-associated lymphomas. MATERIALS AND METHODS: Clinical, histologic, immunophenotypic, and molecular analyses were performed on 103 patients with AIDS lymphomas. RESULTS: Nineteen cases (18.4%) were identified as BLL. They were monoclonal B-cell proliferations, as evaluated by immunoglobulin (Ig) gene rearrangement analyses, and had rearrangement of the c-myc oncogene in 68% of cases but not the bcl-2 gene, in contrast to a previous study on non-HIV associated BLL. This molecular pattern was therefore identical to that of typical BL, suggesting that they represented tumors of similar origin. However, some features could clearly differentiate BLL from BL and were similar to those seen in the diffuse large-cell immunoblastic lymphomas (DLC-IBL) group. These included a greater frequency of Epstein-Barr Virus (EBV) infection (79% v 48%, P = .04), an upregulation of CD39 (50% v 0%, P = .0007) and CD70 (75% v 15%, P = .003) activation antigens and of the CD11a/LFA-1 adhesion molecule (83% v30%, P = .05), and, finally, a lower CD4 count (mean, 119/microL v 270/microL, P = .04). CONCLUSION: BLL is a frequent entity among AIDS lymphomas and should be considered as a morphologic variant of BL in the context of severe immunodepression that occurs in HIV-infected patients. PMID- 9850024 TI - Value of the pretransplant evaluation in predicting toxic day-100 mortality among blood stem-cell and bone marrow transplant recipients. AB - PURPOSE: To determine the value of pretransplant studies in predicting day 100 nonrelapse toxic mortality following high-dose therapy. PATIENTS AND METHODS: A retrospective review of 383 consecutive hematopoietic stem-cell transplants was performed with attention to toxic mortality and pretransplant factors. Univariate log-rank analysis was used to yield the most significant cut-off values for individual factors. Multivariate analysis using Cox proportional hazards regression determined factors independently predictive of early toxic death. RESULTS: Nonrelapse toxic mortality before day 100 occurred in 23 of 383 (6.0%) transplant recipients. Factors associated with an increased risk of toxic death by univariate analysis included forced expiratory volume in 1 second (FEV1) less than 78% of predicted (P = .0002), allogeneic versus autologous transplant (P = .0003), diffusion capacity of carbon monoxide less than 52% of predicted (P = .002), serum creatinine concentration greater than 1.1 mg/dL (P = .003), Eastern Cooperative Oncology Group performance status greater than 0 (P = .006), preparative regimen containing total-body irradiation versus chemotherapy alone (P = .006), marrow versus blood stem cell (P = .01), serum ALT greater than 50 IU/L (P = .02), diagnosis of hematologic disorder versus solid tumor (P = .06), serum bilirubin level greater than 1.1 mg/dL (P = .08), left ventricular ejection fraction (P = .09), and growth factor use (P = .09). In the multivariate model, transplant type (relative risk, 4.2), FEV1 (relative risk, 4.5), performance status (relative risk, 3.7), serum creatinine (relative risk, 3.8), and serum bilirubin (relative risk, 3.7) were found to be independent predictors of early toxic mortality. CONCLUSION: The pretransplant evaluation is a useful tool to identify patients at risk for early toxic mortality following high-dose therapy. PMID- 9850025 TI - Hyper-CVAD and high-dose methotrexate/cytarabine followed by stem-cell transplantation: an active regimen for aggressive mantle-cell lymphoma. AB - PURPOSE: Diffuse and nodular forms of mantle-cell lymphoma (MCL) are consistently associated with poor prognosis. In an effort to improve the outcome, we adopted a treatment plan that consisted of four courses of fractionated cyclophosphamide (CY) 1,800 mg/m2 administered with doxorubicin (DOX), vincristine (VCR), and dexamethasone (Hyper-CVAD) that alternated with high-dose methotrexate (MTX) and cytarabine (Ara-C). After four courses, patients were consolidated with high-dose CY, total-body irradiation, and autologous or allogeneic blood or marrow stem cell transplantation. PATIENTS AND METHODS: Forty-five patients were enrolled; 25 patients were previously untreated, 43 patients had Ann Arbor stage IV disease, and 42 patients had marrow involvement. Forty-one patients had diffuse histology, two patients had nodular, and two patients had blastic variants. RESULTS: Hyper CVAD/MTX-Ara-C induced a response rate of 93.5% (complete response [CR], 38%; partial response [PR], 55.5%) after four cycles of pretransplantation induction chemotherapy. All patients who went on to undergo transplantation achieved CRs. For the 25 previously untreated patients, the overall survival (OS) and event free survival (EFS) rates at 3 years were 92% (95% confidence interval [CI], 80 to 100) and 72% (95% CI, 45 to 98) compared with 25% (95% CI, 12 to 62; P = .005) and 17% (95% CI, 10 to 43; P = .007), respectively, for the previously treated patients. When compared with a historic control group who received a CY, DOX, VCR, and prednisone (CHOP)-like regimen, untreated patients in the study had a 3 year EFS rate of 72% versus 28% (P = .0001) and a better OS rate (92% v 56%; P = .05). Treatment-related death occurred in five patients: all were previously treated and two received allogeneic transplants. CONCLUSION: The Hyper-CVAD/MTX Ara-C program followed by stem-cell transplantation is a promising new therapy for previously untreated patients with MCL. PMID- 9850026 TI - BEACOPP, a new dose-escalated and accelerated regimen, is at least as effective as COPP/ABVD in patients with advanced-stage Hodgkin's lymphoma: interim report from a trial of the German Hodgkin's Lymphoma Study Group. AB - PURPOSE: The HD9 trial aims to evaluate whether moderate dose escalation and/or acceleration of standard polychemotherapy is beneficial for advanced-stage Hodgkin's disease (HD). Two variants of a novel bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone (BEACOPP) scheme (standard and escalated dose) are compared with cyclophosphamide, vincristine, procarbazine, and prednisone (COPP)/doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD). PATIENTS AND METHODS: The randomized, three-arm trial recruited patients in stages IIB and IIIA with risk factors and stages IIIB and IV. BEACOPP in baseline dose contains all drug dosages of COPP/ABVD (except vincristine and procarbazine) rearranged in a shorter, 3-week cycle. Escalated BEACOPP uses higher doses of cyclophosphamide, doxorubicin, and etoposide with granulocyte colony-stimulating factor (G-CSF) support. After eight chemotherapy cycles, initial bulky and residual disease is irradiated. The trial is monitored and analyzed by means of a sequential strategy. RESULTS: An interim analysis with 505 assessable patients and a median follow-up of 23 months showed a significant inferiority (according to sequential monitoring strategy) of the COPP/ABVD regimen in progression rate and freedom from treatment failure (FFTF) compared with the pooled results of both BEACOPP variants. The 24-month FFTF rate was 75% for COPP/ABVD and 84% for BEACOPP pooled (P = .034). There was 12% progressive disease with COPP/ABVD and 6% with BEACOPP pooled. Differences in survival were not significant in sequential analysis. The acute toxicity of baseline BEACOPP resembled that of COPP/ABVD; escalated BEACOPP showed increased but manageable hematologic toxicity. CONCLUSION: Combined with local irradiation, BEACOPP in one or both variants shows superior disease control compared with COPP/ABVD, with acceptable acute toxicity. Further follow-up is required to assess the effect of dosage and the effect on survival and late toxicities. PMID- 9850027 TI - Ongoing somatic mutations and clonal expansions after cure of Helicobacter pylori infection in gastric mucosa-associated lymphoid tissue B-cell lymphoma. AB - PURPOSE: Although most patients with primary gastric low-grade mucosa-associated lymphoid tissue (MALT) B-cell lymphoma experience complete endoscopic and histologic remission after the cure of Helicobacter pylori infection, in many patients, the polymerase chain reaction (PCR) still detects monoclonal B cells in the gastric mucosa. The present study asked whether the lymphoma immunoglobulin VH (IgVH) sequences remained stable in patients with gastric MALT lymphoma after H pylori eradication. PATIENTS AND METHODS: Eight patients with stage EI disease treated with H pylori eradication were analyzed before and at different time points after the cure of the infection. After the amplification of IgVH genes from DNA extracted from gastric biopsy specimens, monoclonal PCR products were cloned and multiple clones (43 to 105) were sequenced per patient. RESULTS: Mutations were detected in all lymphoma VH sequences, which suggested germinal center or postgerminal center origin of the lymphoma B cells. In five of the eight patients, clonal heterogeneity was observed at diagnosis or during follow up. Genealogical analysis of shared and unshared mutations showed that the process of somatic mutations was ongoing after H pylori eradication in four of the five patients who showed clonal instability. Ongoing mutations were observed in three of the four patients who completely responded to H pylori eradication, but in only one of the four patients who did not respond or who partially responded. CONCLUSION: In low-grade gastric MALT lymphomas, an ongoing process of somatic hypermutation and antigen selection can be detected after the therapeutic removal of the underlying stimulus H pylori. These data point to the relevance of yet unknown antigens that drive this disease. In addition, they challenge the view that these lymphomas may be cured solely by the eradication of H pylori. PMID- 9850028 TI - Combination chemotherapy versus melphalan plus prednisone as treatment for multiple myeloma: an overview of 6,633 patients from 27 randomized trials. Myeloma Trialists' Collaborative Group. AB - PURPOSE: To compare combination chemotherapy (CCT) versus melphalan plus prednisone (MP) as treatment for multiple myeloma. PATIENTS AND METHODS: In a collaborative worldwide overview of randomized trials of CCT versus MP, individual patient data on 4,930 patients from 20 trials were analyzed, with the addition of published data on a further 1,703 patients from seven trials. The main outcome measure was mortality, with response and recurrence rates being subsidiary end points. RESULTS: Taking all of the trials together, response rates were significantly higher with CCT than with MP (60.0% v 53.2%; P < .00001, two tailed). There was no evidence of any difference in mortality between CCT and MP, with a nonsignificant 1.5% reduction in death rate in favor of CCT (P = .6, two tailed). There is heterogeneity of design between the trials, but subgroup analyses by type of CCT or by dose-intensities of CCT, of melphalan, or of prednisone did not identify any particular forms of therapy that were either clearly beneficial or clearly adverse. Similarly, analysis of the presentation features of the patients did not find any categories in which CCT differed significantly from MP in its effects on mortality; in particular, there was no evidence that poor-risk patients benefited more from CCT. CONCLUSION: This overview found no difference, either overall or within any subgroup, in mortality between CCT and MP. In terms of survival, these therapeutic options, as tested in the trials considered, are approximately equivalent. PMID- 9850029 TI - Rapid-fractionation preoperative chemoradiation, pancreaticoduodenectomy, and intraoperative radiation therapy for resectable pancreatic adenocarcinoma. AB - PURPOSE: To evaluate the toxicities, radiographic and pathologic responses, and event-free outcomes with combined modality treatment that involves preoperative rapid-fractionation chemoradiation, pancreaticoduodenectomy, and electron-beam intraoperative radiation therapy (EB-IORT) for patients with resectable pancreatic adenocarcinoma. PATIENTS AND METHODS: Patients with radiographically resectable localized adenocarcinoma of the pancreatic head were entered onto a preoperative protocol that consisted of a 2-week course of fluorouracil (5-FU) 300 mg/m2 daily 5 days per week and concomitant rapid-fractionation radiation 30 Gy, 3 Gy daily 5 days per week. Radiographic restaging was performed 4 weeks after chemoradiation, and patients with localized disease underwent pancreaticoduodenectomy with EB-IORT 10 to 15 Gy. RESULTS: Thirty-five patients were entered onto the study and completed chemoradiation, 34 (97%) as outpatients. Three patients (9%) experienced grade 3 nausea and vomiting; no other grade 3 or 4 toxicities were observed. Of the 27 patients taken to surgery, 20 patients (74%) underwent pancreaticoduodenectomy with EB-IORT. All patients had a less than grade III pathologic response to preoperative chemoradiation. At a median follow-up of 37 months, the 3-year survival rate in patients who underwent combined modality therapy was 23%. CONCLUSION: Combined modality treatment with preoperative rapid-fractionation chemoradiation, pancreaticoduodenectomy, and EB-IORT is associated with minimal toxicity and excellent locoregional control. This represents one approach to maximize the proportion of patients who receive all components of combined modality therapy and avoids the toxicity of pancreaticoduodenectomy in patients found to have metastatic disease at the time of restaging. PMID- 9850030 TI - DNA mismatch repair and O6-alkylguanine-DNA alkyltransferase analysis and response to Temodal in newly diagnosed malignant glioma. AB - PURPOSE: We evaluated the response to Temodal (Schering-Plough Research Institute, Kenilworth, NJ) of patients with newly diagnosed malignant glioma, as well as the predictive value of quantifying tumor DNA mismatch repair activity and O6-alkylguanine-DNA alkyltransferase (AGT). PATIENTS AND METHODS: Thirty three patients with newly diagnosed glioblastoma multiforme (GBM) and five patients with newly diagnosed anaplastic astrocytoma (AA) were treated with Temodal at a starting dose of 200 mg/m2 daily for 5 consecutive days with repeat dosing every 28 days after the first daily dose. Immunochemistry for the detection of the human DNA mismatch repair proteins MSH2 and MLH1 and the DNA repair protein AGT was performed with monoclonal antibodies and characterized with respect to percent positive staining. RESULTS: Of the 33 patients with GBM, complete responses (CRs) occurred in three patients, partial responses (PRs) occurred in 14 patients, stable disease (SD) was seen in four patients, and 12 patients developed progressive disease (PD). Toxicity included infrequent grades 3 and 4 myelosuppression, constipation, nausea, and headache. Thirty tumors showed greater than 60% cells that stained for MSH2 and MLH1, with three CRs, 12 PRs, three SDs, and 12 PDs. Eight tumors showed 60% or less cells that stained with antibodies to MSH2 and/or MLH1, with 3 PRs, 3 SDs, and 2 PDs. Eleven tumors showed 20% or greater cells that stained with an antibody to AGT, with 1 PR, 2 SDs, and 8 PDs. Twenty-five tumors showed less than 20% cells that stained for AGT, with 3 CRs, 12 PRs, 4 SDs, and 6 PDs. CONCLUSION: These results suggest that Temodal has activity against newly diagnosed GBM and AA and warrants continued evaluation of this agent. Furthermore, pretherapy analysis of tumor DNA mismatch repair and, particularly, AGT protein expression may identify patients in whom tumors are resistant to Temodal. PMID- 9850031 TI - Phase I clinical and pharmacologic study of weekly cisplatin combined with weekly irinotecan in patients with advanced solid tumors. AB - PURPOSE: In vitro synergy between cisplatin and irinotecan (CPT-11) has been reported. We designed a combination schedule of these agents to maximize the potential for synergistic interaction. PATIENTS AND METHODS: To maximize the opportunity for synergy, we divided the cisplatin into four consecutive weekly treatments, followed by a 2-week rest. Each dose of cisplatin was immediately followed by a dose of irinotecan. The dose of cisplatin was fixed at 30 mg/m2/wk. The initial irinotecan dose was 50 mg/m2/wk and this was escalated by 30% increments in successive cohorts of three to six patients to establish the maximum-tolerated dose (MTD). Pharmacokinetics of irinotecan and its metabolites, SN-38 and SN-38 glucuronide (SN-38G), were analyzed. RESULTS: Of 35 patients with solid tumors enrolled onto this trial, 30 were assessable for toxicity and response. The MTD for this regimen was 30 mg/m2/wk of cisplatin plus 50 mg/m2/wk of irinotecan in previously treated patients and 30 mg/m2/wk of cisplatin plus 65 mg/m2/wk of irinotecan in chemotherapy-naive patients. Neutropenia was the dose limiting toxicity (DLT) encountered in this trial. Diarrhea was infrequent and rarely dose-limiting. Seven of 30 assessable patients achieved a partial response. No alteration in irinotecan, SN-38, or SN-38G pharmacokinetics resulted from the administration of cisplatin with irinotecan. CONCLUSION: The administration of cisplatin and irinotecan on this weekly schedule provides a practical and well-tolerated regimen that has the potential to maximize any clinical synergy between the two agents. Evidence of substantial clinical activity was seen in this phase I study. PMID- 9850032 TI - Phase I study of docetaxel dose escalation in combination with fixed weekly gemcitabine in patients with advanced malignancies. AB - PURPOSE: To determine the maximum-tolerated dose of monthly docetaxel combined with fixed-dose weekly gemcitabine and describe the dose-limiting toxicities (DLTs) of the combination. PATIENTS AND METHODS: Patients with refractory solid tumors were treated with gemcitabine days 1, 8, and 15 every 4 weeks at a fixed dose of 800 mg/m2. Two docetaxel administration schedules were studied, with the drug administered either day 1 or day 15 at doses of 45, 60, 75, and 100 mg/m2 per cycle. RESULTS: Forty patients received 132 cycles of chemotherapy. On the day-1 schedule, the maximum-tolerated docetaxel dose was the highest planned dose of 100 mg/m2 with two DLT episodes among 12 patients treated with 34 cycles at this dose level. On the day-15 schedule, delivery of the planned docetaxel doses was not feasible because of thrombocytopenia and hepatic dysfunction. Hematologic toxicities included grade 4 neutropenia in 16 patients, with three episodes of febrile neutropenia; grades 3 to 4 thrombocytopenia in nine patients; and anemia that required RBC transfusions in 10 patients. For patients treated at the highest docetaxel dose level, myelosuppression was not dose limiting and only one of 34 cycles was complicated by febrile neutropenia. The most common nonhematologic toxicities were asthenia, flu-like symptoms, and fluid retention. Antineoplastic activity was noteworthy, with partial responses in nine of 21 patients with pretreated non-small-cell lung cancer (NSCLC; 43%; 95% confidence interval, 22 to 66), in four of seven patients with breast cancer, and in one patient with esophageal adenocarcinoma. CONCLUSION: Gemcitabine 800 mg/m2 days 1,8, and 15 can be safely combined with docetaxel 100 mg/m2 day 1 of a 28-day cycle. The observed antitumor activity warrants phase II evaluation. PMID- 9850033 TI - Low-dose versus standard-dose lenograstim prophylaxis after chemotherapy: a randomized, crossover comparison. AB - PURPOSE: Granulocyte colony-stimulating factor (G-CSF) administered prophylactically after chemotherapy reduces the duration and severity of neutropenia. This randomized crossover study was designed to assess whether a lower dose of G-CSF is as effective as a standard dose of 5 microg/kg daily. PATIENTS AND METHODS: Patients who received standard-dose chemotherapy regimens expected to cause neutropenia received G-CSF (lenograstim) that started the day after chemotherapy for 14 days or until the absolute neutrophil count (ANC) recovered to greater than 10 x 10(9)/L. The lenograstim dose was randomly allocated to be 2 or 5 microg/kg daily in the first cycle of chemotherapy and crossed over to the alternate dose for the second cycle. The study was designed to accrue 40 assessable patients to provide a power of 80% to detect a difference in duration of neutropenia of 1 day. Fifty-two patients were randomized to treatment and 43 patients completed two cycles of identical chemotherapy. RESULTS: There was little neutropenia irrespective of the dose used. Twenty-three patients (53%) had no grade III or IV neutropenia and 30 patients (70%) had no grade IV neutropenia. Crossover trial methodology was used to assess the difference in outcome caused by the lower dose compared with the standard dose (estimated treatment effect). There was no significant difference in the measures of neutropenia, hospitalization, or other clinical outcomes. The 95% confidence interval (one-sided) for the additional duration of neutropenia caused by the lower dose of lenograstim was 0.43 days or less for grade III or IV neutropenia and 0.34 days or less for grade IV neutropenia. CONCLUSION: Lenograstim 2 microg/kg provides similar protection to 5 microg/kg against neutropenia that complicates standard-dose chemotherapy. The use of a lower dose has important implications for the cost-effectiveness of prophylactic G-CSF therapy. PMID- 9850034 TI - Neuroblastoma and treatment-related myelodysplasia/leukemia: the Memorial Sloan Kettering experience and a literature review. AB - PURPOSE: To assess treatment-related myelodysplasia/leukemia (t-AML) in neuroblastoma patients by a review of the Memorial Sloan-Kettering Cancer Center (MSKCC) data and the literature. PATIENTS AND METHODS: We studied 380 previously untreated and treated MSKCC patients. Low-risk patients received no cytotoxic therapy. High-risk patients received the N4, N5, or N6 regimens. Dosing per cycle and cumulative dosing of leukemogenic agents peaked with N6, which included four cycles of cyclophosphamide 4,200 mg/m2 and doxorubicin 75 mg/m2, plus three cycles of cisplatin 200 mg/m2 and etoposide 600 mg/m2. We reviewed the literature. RESULTS: t-AML occurred in six MSKCC patients, which included three of 53 patients in whom the only chemotherapy consisted of N6, and three patients treated for relapsed or refractory neuroblastoma; no case of leukemia emerged among the 50 low-risk patients. Four cases were found incidentally in routine follow-up bone marrow tests. The 36-month cumulative incidence of t-AML in the N6 cohort was 7% (95% confidence interval, 0 to 15). Published data parallel the MSKCC experience in that t-AML after neuroblastoma was once rare but has become less so since the mid-1980s, when the intensified use of topoisomerase-II inhibitors and alkylators first gained wide acceptance and produced better response rates and longer survival. CONCLUSION: Neuroblastoma itself is not associated with a host susceptibility to leukemia. However, current neuroblastoma treatment programs that use high-dose cyclophosphamide, cisplatin, and topoisomerase-II inhibitors may entail a considerable risk for t-AML. The incidence of t-AML in neuroblastoma patients may be underestimated because treatment and clinical factors can mask its presence. Efforts to devise effective but less leukemogenic treatment for neuroblastoma or to truncate leukemogenic therapy, eg, by exploiting molecular techniques for the early identification of complete remission, are warranted. PMID- 9850035 TI - Current use of bisphosphonates in oncology. International Bone and Cancer Study Group. AB - PURPOSE: The purpose of this article is to review the recent data on bisphosphonate use in oncology and to provide some guidelines on the indications for their use in cancer patients. DESIGN: The group consensus reached by experts on the rationale for the use of bisphosphonates in cancer patients and their current indications for the treatment of tumor-induced hypercalcemia and metastatic bone pain in advanced disease and for the prevention of the complications of multiple myeloma and of metastatic bone disease are reviewed. RESULTS: Bisphosphonates are potent inhibitors of tumor-induced osteoclast mediated bone resorption. They now constitute the standard treatment for cancer hypercalcemia, for which we recommend a dose of 1,500 mg of clodronate or 90 mg of pamidronate; the latter compound is more potent and has a longer lasting effect. Intravenous bisphosphonates exert clinically relevant analgesic effects in patients with metastatic bone pain. Regular pamidronate infusions can also achieve a partial objective response by conventional International Union Against Cancer criteria and enhance the objective response rate to chemotherapy. In breast cancer, the prolonged administration of oral clodronate 1,600 mg daily reduces the frequency of morbid skeletal events by more than one fourth, whereas monthly pamidronate infusions of 90 mg for only 1 year in addition to chemotherapy reduce by more than one third the frequency of all skeletal-related events. The use of bisphosphonates to prevent bone metastases remains experimental. Last, bisphosphonates in addition to chemotherapy are superior to chemotherapy alone in patients with stages II and III multiple myeloma and can reduce the skeletal morbidity rate by approximately one half. CONCLUSION: Bisphosphonate use is a major therapeutic advance in the management of the skeletal morbidity caused by metastatic breast cancer or multiple myeloma, although many questions remain unanswered, notably regarding the optimal selection of patients and the duration of treatment. PMID- 9850036 TI - Economic issues in lung cancer: a review. AB - PURPOSE: Lung cancer is a major source of morbidity, mortality, and health care costs in the developed and developing world. It is estimated that lung cancer is responsible for 20% of all cancer care costs. Concerns exist that this expenditure is associated with questionable benefits. DESIGN: The economic literature that relates to smoking was reviewed, followed by a summary of the economics of the diagnosis, treatment, and palliation of lung cancer. Methodologic considerations are also discussed in this section. RESULTS: Published studies suggest that the increased lifetime health care costs from smoking-related illnesses in smokers are partially or fully offset by the higher medical costs that result from increased longevity in nonsmokers. However, lost productivity costs, which result from morbidity and early mortality among smokers, result in an overall net cost of smoking to society. Discounting rates of 3% to 5% do not substantively alter these results. The per-patient cost to treat lung cancer is substantial. The major cost center is hospitalization; palliative or terminal treatment is associated with significant costs. Savings can be obtained through the judicious use of diagnostic and staging procedures. Furthermore, combined modality treatment approaches and the palliative use of combination chemotherapy appear to be associated with acceptable cost effectiveness compared with commonly used therapies for other diseases. CONCLUSION: Although the increased medical care costs of treating smoking-related diseases are somewhat offset by the higher medical care costs due to increased longevity in nonsmokers, the lost productivity that results from smoking results in a net cost to society. Standard approaches to the management of lung cancer are associated with cost-effectiveness similar to that of other commonly used medical interventions. PMID- 9850037 TI - Primary cardiac angiosarcoma presenting as a malignant pericardial effusion. PMID- 9850039 TI - Adjuvant chemotherapy with CEF versus CMF for node-positive breast cancer. PMID- 9850038 TI - CD20-negative relapse in B-cell lymphoma after treatment with Rituximab. PMID- 9850040 TI - Concurrent G-CSF and chemotherapy. PMID- 9850041 TI - Second cancer after acute lymphoblastic leukemia. PMID- 9850042 TI - Elective internal mammary node irradiation: it is not an all or nothing question. PMID- 9850043 TI - Glutamine treatment of paclitaxel-induced myalgias and arthralgias. PMID- 9850044 TI - Angiogenesis in the hollow fiber tumor model influences drug delivery to tumor cells: implications for anticancer drug screening programs. AB - The National Cancer Institute uses the hollow fiber assay as part of its screening program for anticancer drug discovery. Angiogenesis to hollow fibers implanted s.c. has not been reported, thereby raising concerns about the efficiency of drug delivery and its subsequent effects on chemosensitivity. By extending postimplantation times beyond the 6-day period presently used, extensive vascular networks develop, resulting in both increased delivery and chemosensitivity to doxorubicin. This study suggests that present protocols used to evaluate compounds may produce false negative results, and additional studies to determine the predictive value of the assay are required. PMID- 9850045 TI - Pathogenesis of adenocarcinoma in Peutz-Jeghers syndrome. AB - Peutz-Jeghers syndrome (PJS) is an autosomal dominant condition characterized by intestinal hamartomatous polyps, mucocutaneous melanin deposition, and increased risk of cancer. Families with PJS from the Johns Hopkins Polyposis Registry were studied to identify the molecular basis of this syndrome and to characterize the pathogenesis of gastrointestinal hamartomas and adenocarcinomas in PJS patients. Linkage analysis in the family originally described by Jeghers in 1949 and five other families confirmed linkage to 19p13.3 near a recently identified gene responsible for PJS. Germ-line mutations in this gene, STK11, were identified in all six families by sequencing genomic DNA. Analysis of hamartomas and adenocarcinomas from patients with PJS identified loss of heterozygosity (LOH) of 19p markers near STK11 in 70% of tumors. Haplotype analysis indicated that the retained allele carried a germ-line mutation, confirming that STK11 is a tumor suppressor gene. LOH of 17p and 18q was identified in an adenocarcinoma but not in hamartomas, implying that allelic loss of these two regions corresponds to late molecular events in the pathogenesis of cancer in PJS. The adenocarcinomas showing 17p LOH also demonstrated altered p53 by immunohistochemistry. None of the 18 PJS tumors showed microsatellite instability, LOH on 5q near APC, or mutations in codons 12 or 13 of the K-ras proto-oncogene. These data provide evidence that STK11 is a tumor suppressor gene that acts as an early gatekeeper regulating the development of hamartomas in PJS and suggest that hamartomas may be pathogenetic precursors of adenocarcinoma. Additional somatic mutational events underlie the progression of hamartomas to adenocarcinomas, and some of these somatic mutations are common to the later stages of tumor progression seen in the majority of colorectal carcinomas. PMID- 9850046 TI - Propagation of genetically altered tumor cells derived from fine-needle aspirates of primary breast carcinoma. AB - Because primary breast tumors are diagnosed earlier in the clinic, procurement of sufficient amounts of tumor tissue for in-depth biological characterization is becoming increasingly difficult. We demonstrate here that relatively small numbers of tumor cells within samples of fine-needle aspirates (FNA) can be propagated in culture. Of 25 cases attempted, 12 were passageable, resulting in up to 10(7) viable cells. FNA-derived cultures were evaluated for anchorage independence, c-erb-B2 overexpression, aneusomy, and pattern of allelic loss. In every case examined, the cultured cells closely resembled the original tumor tissue and displayed one or more tumor phenotypes. The incidence of erb-B2 overexpressing tumors was similar in passageable and nonpassageable cases (33% versus 31%, respectively). FNAs that are expanded from a wide range of clinical breast material could be useful for functional studies presently limited to rare established cell lines, such as aberrant signal transduction and gene regulation, and for testing potential anticancer vaccines and drugs. PMID- 9850047 TI - Induction of brain tumors in mice using a recombinant platelet-derived growth factor B-chain retrovirus. AB - In existing mouse models for malignant brain tumors, genes with no proven pathogenical relevance for humans have been used. Coexpression of platelet derived growth factor (PDGF) and PDGF receptors suggests an autocrine mechanism of growth factor stimulation in the development of brain tumors in man. A murine retrovirus coding for the PDGF B-chain was, therefore, used to induce brain tumors in mice. Of 35 mice who received injections, 15 developed brain tumors of oligo- or monoclonal origin. They coexpressed PDGF B-chain and alpha-receptor mRNA, as expected, from an autocrine mechanism of transformation. Most tumors displayed characteristics of glioblastoma multiforme or of a primitive neuroectodermal tumor, and the consistent expression of nestin suggested that they were all derived from an immature neuroglial progenitor. The results show that an autocrine mechanism of transformation may be an initial or early event in neuro-oncogenesis. The present model provides an ideal system for studies of genetic mechanisms involved in the development of brain tumors. PMID- 9850048 TI - Increased expression of hypoxia inducible factor-1alpha in rat and human prostate cancer. AB - Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that regulates genes involved in adaptation to hypoxia. Expression of HIF-1alpha was evaluated in rat and human prostate cancer cell lines. Increased expression of HIF-1alpha mRNA in rat prostate cancer cell lines and hypoxia-induced expression of HIF-1alpha protein in human prostate cancer cell lines are associated with increased cell growth rates and metastatic potential. HIF-1alpha mRNA was undetectable in the normal rat ventral prostate by Northern blot hybridization. HIF-1alpha protein expression and HIF-1 DNA binding activity were detected in normoxic PC-3 cells. Human prostate cancer cells plated at low density manifested higher functional HIF-1alpha expression than cells plated at high density independent of O2 tension. HIF-1alpha may become dysregulated in prostate cancer and thus drive the transcription of hypoxia-adaptive genes involved in tumor progression. This is also the first evidence that human cancer cells can express functional HIF-1alpha protein under normoxic conditions. PMID- 9850049 TI - Adenoviral transgene expression of MMAC/PTEN in human glioma cells inhibits Akt activation and induces anoikis. AB - The MMAC/PTEN tumor suppressor gene encodes for a phosphatase that recently has been shown to have phosphotidylinositol phosphatase activity, implicating its possible involvement in phosphatidylinositol 3'-kinase-mediated signaling. To investigate possible alterations in growth factor-mediated signal transduction, an adenovirus containing MMAC/PTEN, Ad-MMAC, previously shown to inhibit growth and tumorigenicity in glioma cells, was used to acutely express the transgene. Human glioma cells infected with Ad-MMAC but not with control adenoviruses exhibited an inhibition of phosphorylation of both activating residues of Akt, Ser-473, and Thr-308, along with Akt's serine/threonine kinase activity, without significantly altering Akt expression. The effects of functional MMAC/PTEN expression were relatively specific, because members of several other growth factor-mediated signaling pathways showed no altered responses. The presence of MMAC/PTEN also inhibited phosphorylation of BAD, although no evidence of apoptosis in the in situ treated cells was observed. However, U251 glioma cells infected with Ad-MMAC were induced to undergo anoikis at a significantly higher rate than U251 cels treated with control viruses or mock infected with media. These results demonstrate that the acute administration of MMAC/PTEN results in the inhibition of Akt-mediated signaling, growth inhibition, and anoikis, implying that loss of MMAC/PTEN increases cellular proliferation and significantly augments a cell's survival potential during cellular processes that are associated with malignancy. PMID- 9850050 TI - Association between lung cancer and microsomal epoxide hydrolase genotypes. AB - Microsomal epoxide hydrolase (mEH) is involved in the metabolism of tobacco derived carcinogens. Polymorphisms in exons 3 and 4 of the EPHX gene have been reported to be associated with variations in mEH activity. We examined whether the predicted mEH activity modified the lung cancer risk among 150 cases and 172 controls, all French Caucasian smokers. A significant association was found between predicted mEH activity and lung cancer (P < 0.02), with a dose-effect relationship (P < 0.005). The risks associated with intermediate and high activities, compared to low activity, were 1.65 (95% CI, 0.95-2.86) and 2.66 (95% CI, 1.33-5.33), respectively. The effect of mEH activity on lung cancer risk was not significantly modified by smoking exposure, CYP1A1 genotype, or GSTM1 genotype. mEH may thus be an important genetic determinant of smoking-induced lung cancer. PMID- 9850051 TI - The bisphosphonate incadronate (YM175) causes apoptosis of human myeloma cells in vitro by inhibiting the mevalonate pathway. AB - It has recently been suggested that bisphosphonates may have direct antitumor effects in vivo, in addition to their therapeutic antiresorptive properties. Bisphosphonates can inhibit proliferation and cause apoptosis in human myeloma cells in vitro. In macrophages, bisphosphonate-induced apoptosis was recently found to be a result of inhibition of the mevalonate (MVA) pathway. The aim of this study was to determine whether bisphosphonates also affect human myeloma cells in vitro by inhibiting the MVA pathway. Incadronate and mevastatin (a known inhibitor of the MVA pathway) caused apoptosis in JJN-3 myeloma cells and inhibited cell proliferation. Geranylgeraniol and farnesol prevented incadronate induced apoptosis and had a partial effect on cell cycle arrest. MVA and geranylgeraniol prevented mevastatin-induced apoptosis and inhibition of proliferation and completely prevented the effect of mevastatin on the cell cycle. These observations demonstrate that incadronate-induced apoptosis in human myeloma cells in vitro is the result of inhibition of the MVA pathway. PMID- 9850052 TI - Absence of topoisomerase IIbeta in an amsacrine-resistant human leukemia cell line with mutant topoisomerase IIalpha. AB - Numerous chemotherapeutic agents act via stabilization of a topoisomerase (topo) II-DNA complex. HL-60/AMSA, a human leukemia cell line, is resistant to intercalator-mediated DNA complex formation and cytotoxicity. HL-60/AMSA contains a mutant form of topo IIalpha that was thought to explain this resistance. However, our present data show that expression of topo IIbeta RNA in HL-60/AMSA is only 10% of that in HL-60, and topo IIbeta protein levels are undetectable. Southern analysis of topo IIbeta shows no differences in gene dosage between the two cell lines but does show differences in the restriction patterns. These data suggest that decreased topo IIbeta expression may contribute to the intercalator resistance of HL-60/AMSA cells. PMID- 9850054 TI - Generation of CD8+ and CD4+ T-cell response to dendritic cells genetically engineered to express the MART-1/Melan-A gene. AB - Both CD8+ and CD4+ T cells have demonstrated roles in antitumor immune response in many animal tumor systems. In many human tumor systems, although abundant literature exists on the evidence of tumor antigen-specific CD8+ CTL response, only limited information is available on tumor antigen-specific CD4+ T-cell response. Using the MART-1/Melan-A (MART-1) antigen system as a prototype human tumor-associated antigen (TAA)- and dendritic cell (DC)-based MART-1 antigen presentation system (i.e., DCs transduced with an adenoviral vector-based construct carrying the MART-1 gene), we explored, in vitro, the feasibility of generating both CD8+ and CD4+ T-cell responses in the same individual. Here, we show that autologous DCs from both HLA-A2-positive melanoma patients and normal healthy individuals that are transduced with an adenoviral vector containing the MART-1 antigen are capable of inducing both MART-1-specific CD8+ and CD4+ T cells in in vitro coculture. After several rounds of stimulation, both the CD4+ and CD8+ T cells synthesized IFN-gamma when they were specifically stimulated. The CD8+ T cells generated in such cocultures also recognized the MART-1(27-35) peptide, AAGIGILTV, in 4-h cytotoxicity assays. These observations, therefore, suggest that Th1-type responses can be generated, in vitro, by stimulation with DCs that are genetically modified to express a TAA. Although the outcome of this type of genetically engineered DC-based stimulation may vary from system to system, this type of in vitro antigen presentation may be very useful in more comprehensive analyses of CD4+ T-cell response to defined TAAs, and such genetically engineered autologous DCs might be better candidates to serve as surrogate cancer vaccines. PMID- 9850053 TI - Realization of the therapeutic potential of CTLA-4 blockade in low-dose chemotherapy-treated tumor-bearing mice. AB - CTLA-4 blockade has been shown by other investigators [D. R. Leach, et al., Science (Washington DC), 271: 1734-1736, 1996; and Y-F. Yang, et al., Cancer Res., 57: 4036-4041, 1997] to retard tumor growth in selected tumor systems. Here, we show that CTLA-4 blockade alone was ineffective in retarding tumor growth in the murine MOPC-315 tumor system. Yet, CTLA-4 blockade offered significant therapeutic benefits to MOPC-315 tumor bearers when combined with a subtherapeutic dose of the chemotherapeutic agent melphalan, which was previously shown (L. Gorelik, et al., Cancer Immunol. Immunother., 39: 117-126, 1994) to shift the cytokine profile in the tumor bearers toward type-1 cytokines. In addition, we show here that anti-CTLA-4 monoclonal antibody enhanced antitumor cytotoxicity when the anti-CTLA-4 monoclonal antibody was added to stimulation cultures of spleen cells from low-dose melphalan-treated MOPC-315 tumor-bearing mice but not from untreated tumor-bearing mice. These results suggest that the therapeutic benefits of CTLA-4 blockade depend on the ability of drugs such as melphalan to promote an immunogenic environment by altering the cytokine profile of tumor-specific T cells. PMID- 9850055 TI - Methylation of the androgen receptor promoter CpG island is associated with loss of androgen receptor expression in prostate cancer cells. AB - Androgen-independent metastatic prostate cancer is characterized by a heterogeneous loss of androgen receptor (AR) expression among tumor cells. In this study, we evaluate DNA hypermethylation as a potential transcriptional regulatory mechanism in AR-negative prostate cancer cell lines. Nucleotide sequence analysis demonstrates an approximately 15-kb CpG island in the AR gene that encompasses the transcription start site and exon 1. Using Southern blotting with methylation-sensitive restriction enzymes and methylation-specific PCR, we find aberrant methylation in the AR expression-negative cell lines Du145, DuPro, TSU-PR1, and PPC1. Incomplete methylation in the AR CpG island is also seen in normal female breast and ovarian tissues consistent with the inactivation of one X chromosome by hypermethylation. In contrast, prostate cancer cell lines LNCaP and PC3 express AR and are unmethylated. Normal prostate epithelial cell strains demonstrate no methylation. Exposure of AR-negative prostate cancer cell lines to 5-aza-2' deoxycytidine, a demethylating agent, induces the reexpression of AR RNA in DuPro and TSU-PR1. This reexpression is associated with a demethylation of this region. Prostate-specific antigen, an androgen-responsive gene, is also specifically induced in these lines after AR reexpression. Therefore, in vitro DNA methylation of the 5' CpG AR island may be associated with the loss of AR expression. Furthermore, our results demonstrate that treatment with demethylating agents may engender the reexpression and function of the androgen receptor in AR-negative cell lines. PMID- 9850056 TI - IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs. AB - Survivin is a member of the inhibitor of apoptosis protein (IAP) family. We investigated the antiapoptotic mechanism of Survivin, as well as its expression in 60 human tumor cell lines used for the National Cancer Institute's anticancer drug screening program. In cotransfection experiments, cell death induced by Bax or Fas (CD 95) was partially inhibited (mean +/- SD, 65% +/- 8%) by Survivin, whereas XIAP, another IAP family member, almost completely blocked cell death (93% +/- 4%) under the same conditions. Survivin and XIAP also protected 293 cells from apoptosis induced by overexpression of procaspase-3 and -7 and inhibited the processing of these zymogens into active caspases. In vitro binding experiments indicated that, like other IAP-family proteins, Survivin binds specifically to the terminal effector cell death proteases, caspase-3 and -7, but not to the proximal initiator protease caspase-8. Using a cell-free system in which cytosolic extracts were derived from control- or Survivin-transfected cells and where caspases were activated either by addition of cytochrome c and dATP or by adding recombinant active caspase-8, Survivin was able to substantially reduce caspase activity, as measured by cleavage of a tetrapeptide substrate, AspGluValAsp-aminofluorocoumarin. Similar results were obtained in intact cells when Survivin was overexpressed by gene transfection and caspase activation was induced by the anticancer drug etoposide. Survivin was expressed in all 60 cancer cell lines analyzed, with highest levels in breast and lung cancers and lowest levels in renal cancers. These findings indicate that Survivin, which is commonly expressed in human tumor cell lines, can bind the effector cell death proteases caspase-3 and -7 in vitro and inhibits caspase activity and cell death in cells exposed to diverse apoptotic stimuli. Although quantitative differences may exist, these observations suggest commonality in the mechanisms used by IAP family proteins to suppress apoptosis. PMID- 9850057 TI - A single nucleotide polymorphism in the matrix metalloproteinase-1 promoter creates an Ets binding site and augments transcription. AB - Matrix metalloproteinases (MMPs) facilitate cellular invasion by degrading the extracellular matrix, and their regulation is partially dependent on transcription. Binding sites for members of the Ets family of transcription factors are present within MMP promoters and are potent positive regulators. We report a single nucleotide polymorphism at -1607 bp in the MMP-1 promoter, where an additional guanine (G) creates an Ets binding site, 5'-GGA-3'. This polymorphism displays significantly higher transcription in normal fibroblasts and in melanoma cells than the 1 G polymorphism, and it binds substantially more nuclear extract and recombinant ETS-1. Analysis of control DNAs from the Center d'Etude du Polymorphisme Humain pedigrees reveals that this polymorphism is not a mutation, with a frequency of the 2 G polymorphism at 30%. In contrast, in eight tumor cell lines, this frequency increased to 62.5% (P < 0.0001). Thus, this MMP 1 polymorphism contributes to increased transcription, and cells expressing the 2 G polymorphism may provide a mechanism for more aggressive matrix degradation, thereby facilitating cancer progression. PMID- 9850058 TI - An improved method for construction of directionally cloned cDNA libraries from microdissected cells. AB - Here, we developed an improved method for constructing microdissected cDNA libraries, based on strand-switching properties of reverse transcriptase, followed by PCR amplification with primers to mediate unidirectional insert cloning. Using RNA from microdissected ovarian carcinoma cells, we constructed a cDNA library consisting of 1.3 x 10(6) unidirectional recombinants with an average insert size of 500 bp. Single-pass sequencing of 100 clones with the T7 primer revealed 89 inserts derived from known genes, anonymous expressed sequence tags (ESTs), or novel sequences. Among these clones were known genes and ESTs previously found in cDNA libraries from bulk ovarian tissue RNA, sequences seen for the first time in an ovarian-derived library, and novel sequences not previously seen in any cDNA library. These results demonstrate a methodology for constructing quality cDNA libraries that are cloned in a unidirectional fashion, are complex and diverse, and reflect the tissue of origin. PMID- 9850059 TI - Genetic alterations of the transforming growth factor beta receptor genes in pancreatic and biliary adenocarcinomas. AB - Transforming growth factor beta (TGF-beta) is an extracellular ligand that binds to a heterodimeric receptor, initiating signals that regulate growth, differentiation, and apoptosis. Many cancers, including pancreatic cancer, harbor defects in TGF-beta signaling and are resistant to TGF-beta-mediated growth suppression. Genetic alterations of DPC4, which encodes a DNA binding protein that is a downstream component of the pathway, most frequently occur in pancreatic and biliary carcinomas. We searched for other targets of mutation of the TGF-beta pathway in these cancers. We report somatic alterations of the TGF beta type I receptor gene ALK-5. Homozygous deletions of ALK-5 were identified in 1 of 97 pancreatic and 1 of 12 biliary adenocarcinomas. A germ-line variant of ALK-5, presumably a polymorphism, was identified, but no somatic intragenic mutations were identified upon sequencing of all coding regions of ALK-5. Somatic alterations of the TGF-beta type II receptor gene (TGFBR2) were identified in 4 of 97 (4.1%) pancreas cancers, including a homozygous deletion in a replication error-negative cancer and three homozygous frameshift mutations of the poly(A) tract of the TGF-beta type II receptor in replication error-positive cancers. We also studied other related type I receptors of the TGF-beta superfamily. In a panel of pancreas cancers preselected for loss of heterozygosity at the ALK-1 locus, sequencing of all coding exons of the ALK-1 gene revealed no alterations. No homozygous deletions were detected in the ALK-1, ALK-2, ALK-3, or ALK-6 genes in a panel of 86 pancreatic cancer xenografts and 11 pancreatic cancer and 22 breast cancer cell lines. The rate of genetic inactivation of TGF-beta pathway members was determined in 45 pancreatic cancers. Eighty-two % of these pancreatic cancers had genetic inactivation of the DPC4, p15, ALK-5, or TGFBR2 genes. Our results indicate that the TGF-beta type I and type II receptor genes are selective targets of genetic inactivation in pancreatic and biliary cancers. PMID- 9850060 TI - Inactivation of the p53 tumor suppressor gene via a novel Alu rearrangement. AB - Inactivation of the p53 tumor suppressor gene is a common finding in human cancer. In most cases, inactivation is due to a point mutation in the gene, but rearrangement of the p53 gene is sometimes observed. We analyzed the inactivation of p53 in the human pancreas cancer cell line Hs766T, which harbors a structural alteration in the p53 gene. This inactivation was found to be the result of a complex deletion/insertion event involving at least two different Alu elements. The rearrangement eliminated exons 2-4 from the p53 gene, whereas a 175-bp Alu fragment was inserted between the breakpoints of the deletion. DNA sequence analysis of this Alu fragment revealed that it is identical to an Alu element in intron 1 of the p53 gene. This is the first report of p53 inactivation due to a rearrangement involving Alu elements. This type of inactivation may go unnoticed when only traditional methods to detect p53 alterations are used. PMID- 9850061 TI - A human placenta-specific ATP-binding cassette gene (ABCP) on chromosome 4q22 that is involved in multidrug resistance. AB - We characterized a new human ATP-binding cassette (ABC) transporter gene that is highly expressed in the placenta. The gene, ABCP, produces two transcripts that differ at the 5' end and encode the same 655-amino acid protein. The predicted protein is closely related to the Drosophila white and yeast ADP1 genes and is a member of a subfamily that includes several multidrug resistance transporters. ABCP, white, and ADP1 all have a single ATP-binding domain at the NH2 terminus and a single COOH-terminal set of transmembrane segments. ABCP maps to human chromosome 4q22, between the markers D4S2462 and D4S1557, and the murine gene (Abcp) is located on chromosome 6 28-29 cM from the centromere. ABCP defines a new syntenic segment between human chromosome 4 and mouse chromosome 6. The abundant expression of this gene in the placenta suggests that the protein product has an important role in transport of specific molecule(s) into or out of this tissue. PMID- 9850062 TI - Catalytic efficiencies of allelic variants of human glutathione S-transferase P1 1 toward carcinogenic anti-diol epoxides of benzo[c]phenanthrene and benzo[g]chrysene. AB - Four allelic variants of glutathione (GSH) S-transferase P1-1 (hGSTP1-1) that differ in their structures at amino acid(s) in position(s) 104 and/or 113 are known to exist in human populations. However, the physiological significance of hGSTP1-1 polymorphism is not fully understood. In this communication, we report that the I104,A113 allele of hGSTP1-1, which is most frequent in human populations, is also most efficient in the GSH conjugation of carcinogenic anti diol epoxides of benzo[g]chrysene and benzo[c]phenanthrene (anti-BGCDE and anti BCPDE, respectively). The catalytic efficiency of hGSTP1-1(I104,A113) isoform toward anti-BGCDE, 0.36 mM(-1) x s(-1), was approximately 1.7-fold higher (P < 0.05) compared with hGSTP1-1(V104,V113). Interestingly, the frequency of codon 104-valine alleles is significantly higher in certain cancers compared with codon 104-isoleucine alleles. Like anti-BGCDE, the catalytic efficiency of hGSTP1 1(I104,A113) isoform toward anti-BCPDE was higher by about 1.4- to 2.2-fold (P < 0.05) than those of other hGSTP1-1 variants. These observations are interesting because we have shown previously (Hu, X. et al., Biochem. Biophys. Res. Commun., 238: 397-402, 1997) that the V104,V113 variant, not the I104,A113 isoform, is most efficient in the GSH conjugation of bay-region anti-diol epoxide of benzo(a)pyrene (anti-BPDE), which, unlike anti-BGCDE or anti-BCPDE, is a planar molecule. In conclusion, our results suggest that hGSTP1-1 polymorphism may be an important factor in differential susceptibility of humans to cancers where polycyclic aromatic hydrocarbons are etiological factors and that I104,A113 variant may play a major role in the detoxification of nonplanar, sterically hindered fjord-region diol epoxides (e.g., anti-BGCDE). PMID- 9850063 TI - Absent or low expression of the zeta chain in T cells at the tumor site correlates with poor survival in patients with oral carcinoma. AB - Immunohistology for expression of the CD3zeta and CD3epsilon chains in TIL was performed in 138 paraffin-embedded primary oral squamous cell carcinoma tissues and 10 nontumor, inflammatory lesions. Semiquantitative analysis of the staining intensity for zeta chain expression and number of zeta chain expression-positive cells distinguished tumors with absent or low zeta expression (42 of 132) from those with normal zeta expression (90 of 132). Zeta chain expression was inversely correlated with the tumor stage. Survival was significantly lower in patients whose TIL had absent or low zeta expression, controlling for stage (P = 0.003) and lymph node status (P = 0.0005). The prognostic value of zeta chain was restricted to patients with stage III or IV tumors (P = 0.003). The data indicate that absent or decreased zeta expression in TIL combined with tumor stage or nodal status defines a group of patients with oral squamous cell carcinoma who have an extremely poor prognosis. PMID- 9850064 TI - Presence and location of TP53 mutation determines pattern of CDKN2A/ARF pathway inactivation in bladder cancer. AB - Transformation and immortalization require the inactivation of key cell cycle regulatory genes. We examined 19 bladder cancer cell lines derived from 17 patients for alterations in TP53, RB1, CDKN2A, and ARF. Twelve cell lines had a mutation in exons 5-11 of TP53 and, with only one exception, a concomitant loss of RB1 protein expression. Another group of seven cell lines had a wild-type TP53 gene or a mutation in exons 1-4 of TP53 and concomitant alterations in both CDKN2A and ARF in every case. This demonstrates the requirement, in all but one line, for inactivation of both the CDKN2A/RB1 and ARF/TP53 pathways in bladder cancer cell lines and provides the first evidence for potential differences in the penetrance of mutations in the transactivation and DNA-binding domains of TP53. PMID- 9850065 TI - Prevention of NNK-induced lung tumorigenesis in A/J mice by acetylsalicylic acid and NS-398. AB - Acetylsalicylic acid (ASA) is known to prevent cancer development, but its mechanism of action remains unclear. In this study, we compared the efficacies of this nonspecific cyclooxygenase (COX) inhibitor with N-[2-(cyclohexyloxy)-4 nitrophenyl]-methanesulfonamide (NS-398), a specific COX-2 inhibitor. COX-2 specific inhibitors are less toxic than ASA. Lung tumorigenesis was induced in A/J mice by the administration of the tobacco-specific nitrosamine 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in the drinking water for 7 weeks (weeks 0 to +7). Groups of 25 A/J mice were fed ASA (588, 294, 147, or 73 mg/kg diet) before and throughout the assay (weeks -2 to +23). ASA at a dose of 588 mg/kg diet was the most effective because it reduced lung tumor multiplicity by 53%. The preventive effect of ASA increased with the dose, being of 32, 30, and 44% for 73, 147, and 294 mg/kg diet, respectively. NNK increased plasma prostaglandin E2 (PGE2) basal levels by 413%, whereas ASA attenuated this elevation in a dose-response manner (r2 = 0.99). Plasma PGE2 levels in ASA + NNK treated mice correlate with the logarithm of the number of tumors (r2 = 0.99). NS 398 inhibited lung tumor multiplicity by 34% and returned plasma PGE2 to basal levels observed in untreated mice. Among the NNK-exposed mice, ASA and NS-398 treatment decreased the mean of the lung tumor volumes. Incubation of 82-132 and LM2 murine lung tumor cells with ASA or NS-398 decreased cell proliferation by 50% at concentrations higher than 100 microM. Incubations of NNK with COX-1 and 2 produced both activation and detoxification products by alpha-carbon hydroxylation and N-oxydation pathways, respectively. Bioactivation of NNK was more extensive by COX-2 than COX-1. Anti-COX-1 and -2, arachidonic acid, ASA, and NS-398 inhibited NNK bioactivation by COX-1 and -2 from 22-49%. Our data suggest that NNK is bioactivated by COX-2 in lung tissues and that COX-2-specific inhibitors might be promising chemopreventive agents. PMID- 9850066 TI - The augmentation of melanocytic nevi in guinea pigs by solar-simulated light: an animal model for human melanocytic nevi. AB - Strong epidemiological evidence confirms the role of sunlight in human melanoma induction. Furthermore, the frequency of melanocytic nevi is a good indicator of future development of melanoma and a short-term marker of adverse reactions to melanoma-inducing sun exposure in humans. Thus, the aim of this study was to develop and define an animal model for sunlight-induced nevi that can be used as a surrogate model for sunlight-induced melanoma. Five treatment groups of 30-40 Hartley albino guinea pigs/group were treated with topical 7,12 dimethylbenzanthracene at a dose range of 6-240 mg on the dorsum of the skin. At week 20, half of the animals in each group were given a 12-month regimen of minimal erythemal solar-simulated light, 3 times/week, increased weekly to maintain erythema. These regimes induced epidermally derived pigmented melanocytic nevi clinically and histologically similar to human nevi (junctional, compound, and dermal). S100 and HMB45 staining was also consistent with the patterns seen in human nevi. In contrast to the high-dose 7,12 dimethylbenzanthracene-treated animals (60 and 240 mg), where solar-simulated light had no effect on nevi multiplicity, those groups treated with low doses (24, 12, and 6 mg) had a significant increase in nevi multiplicity after 12 months of solar-simulated light treatment (24 mg, 0.5 nevi/animal unirradiated versus 1.4 nevi/animal irradiated, P = 0.03; 12 mg, 0.2 unirradiated versus 1.2 irradiated, P = 0.02; 6 mg, 0 unirradiated versus 1.9 irradiated, P = 0.008). UVB induced minimal erythemal dose was unaltered after exposure to photoreactivating light, consistent with the observation of others that placental mammals lack the DNA photolyase responsible for strong photoreactivation seen in nonplacental mammals and lower metazoans. Thus, our guinea pig model has some of the essential elements required to be a robust animal model for human nevi and a surrogate model for melanoma. These nevi are augmented by solar-simulated light, are histologically similar, occupy the same level within the skin, have the same natural history as human nevi, and are produced in an animal lacking strong photoreactivation. These features are not found in any previously described small laboratory animal model. PMID- 9850067 TI - Differential expression of estrogen receptor-alpha and -beta messenger RNAs as a potential marker of ovarian carcinogenesis. AB - Although estrogen receptor (ER)-alpha is expressed in both benign and malignant ovarian tumors, the role of ER in ovarian carcinogenesis of epithelial tumors is still unknown. In view of the recent characterization of ER-beta, a second form of ER that seems to be highly expressed in ovaries, we reexamined this issue by studying the relative expression of ER-alpha and -beta in human ovarian tumor progression. We developed a competitive PCR assay based on coamplification of the two ERs in target nucleotide sequences displaying a high homology (exons 3 and 4). Coamplification experiments with varying amounts of plasmids containing ER alpha and -beta cDNAs showed that this assay was reliable for discriminating as little as a 2-fold difference in the initial ER-alpha:ER-beta cDNA ratio. The relative expression of ER-alpha compared with ER-beta mRNAs was studied in human ovarian cancer cell lines (n = 5) and in normal ovaries (n = 6), then in human benign and malignant tumor samples including ovarian cysts (n = 24), borderline tumors (n = 3), and cancers (n = 10). In normal ovaries, ER-beta mRNA was the predominant ER form, whereas in ovarian cancer cell lines ER-alpha mRNA was markedly increased as compared with ER-beta. In benign and borderline tumors, ER beta mRNA was detected in 78% of tumors, whereas ER-alpha mRNA was detected in 29%. In ovarian carcinomas, both ER-alpha and -beta mRNAs were expressed in 80% of tumors. The ER-alpha:ER-beta mRNA ratio was >1 in only one cyst sample (4%). In contrast, the ER-alpha:ER-beta mRNA ratio was markedly increased in ovarian cancers because 60% showed an ER-alpha:ER-beta mRNA >1. In situ hybridization experiments showed overlapping tissular distribution of ER-beta and -alpha expression in cancers and cysts, with a main localization in the epithelium and only a low level of expression in stromal cells. In summary, we found an increase in the ER-alpha:ER-beta mRNA ratio in ovarian carcinomas as compared with normal ovaries and cysts. These data suggest that overexpression of ER-alpha relative to ER-beta mRNA may be a marker of ovarian carcinogenesis. PMID- 9850068 TI - Protection of metabolic and exercise capacity in unselected weight-losing cancer patients following treatment with recombinant erythropoietin: a randomized prospective study. AB - This study was aimed at evaluating whether anemia could be prevented in unselected weight-losing cancer patients on anti-inflammatory treatment by early and prophylactic treatment with recombinant human erythropoietin (rhEPO) and whether such a benefit could be translated into improved physical function and metabolic efficiency. One hundred eight cancer patients who experienced progressive cachexia due to solid, mainly gastrointestinal tumors were randomized to receive twice daily a cyclo-oxygenase inhibitor (controls; indomethacin, 50 mg twice a day) or indomethacin and erythropoietin, provided on individual basis to prevent development of progressive anemia (study patients; indomethacin, 50 mg twice a day plus rhEPO; range, 12,000-30,000 units per week). All patients were treated and followed up until death or to preterminal stage. Biochemical tests (blood, liver, kidney, and thyroid), nutritional state assessment (food intake and body composition), and exercise testing with simultaneous measurements of respiratory gas exchanges before and during exercise were performed before institution of treatments and then at regular intervals during the treatment period (2-30 months after start). Study and control patients did not differ in survival. rhEPO prevented development of anemia during the entire observation period. This was associated with a significantly more preserved maximum exercise capacity in study patients compared to control patients during the follow-up period (101 +/- 10 versus 66 +/- 6 W; P < 0.0001), based on more effective ventilation and whole-body respiratory gas exchanges. These improvements were also evident when exercise performance was normalized to lean body mass, an indirect measure of the skeletal muscle mass. The metabolic efficiency, expressed as oxygen uptake per watt produced, was also significantly preserved in rhEPO treated patients compared to controls (14.1 +/- 1.1 versus 16.3 +/- 0.9 ml O2/W, P < 0.05). Our results demonstrate that institution of early and prophylactic rhEPO treatment to patients with progressive cancer prevents development of tumor induced anemia. This achievement was associated with a better preserved exercise capacity, which is explained in part by improved whole-body metabolic and energy efficiency during work load. PMID- 9850069 TI - Alpha-difluoromethylornithine inhibits N-nitrosomethylbenzylamine-induced esophageal carcinogenesis in zinc-deficient rats: effects on esophageal cell proliferation and apoptosis. AB - Sustained, increased cell proliferation induced by dietary zinc deficiency in rats plays a critical role in esophageal carcinogenesis. It is the determining factor that converts an otherwise nontumorigenic dose of N nitrosomethylbenzylamine (NMBA) into a highly tumorigenic one. We studied whether the increased esophageal cell proliferation and susceptibility to NMBA-induced carcinogenesis induced by zinc deficiency can be inhibited by alpha difluoromethylornithine (DFMO), an enzyme-activated, irreversible inhibitor of ornithine decarboxylase (the first enzyme in polyamine synthesis). Weanling rats were divided into four groups: Zn+/DFMO-, Zn+/DFMO+, Zn-/DFMO-, and Zn-/DFMO+. They were fed ad libitum either a zinc-sufficient (Zn+, 75 ppm zinc) or a zinc deficient (Zn-, 4 ppm zinc) diet and given either deionized water (DFMO-) or 1% DFMO in deionized water (DFMO+). After 5 weeks, 5-19 animals from each group were sacrificed after in vivo 5-bromo-2'-deoxyuridine labeling to detect cells in S phase. The remaining animals in each group were given a single intragastric dose of NMBA at 2 mg/kg and sacrificed 12 weeks later for tumor incidence analysis. At week 5, DFMO treatment greatly decreased (by 48-82%) the levels of putrescine and spermidine in rat esophagus, colon, and liver, irrespective of dietary zinc intake. The increased esophageal cell proliferation induced by dietary zinc deficiency, as measured by the labeling index, the number of labeled cells, and the total number of cells, was substantially reduced by DFMO. This was accompanied by an increase in the rate of apoptosis. In addition, the expression of bax protein, an apoptosis accelerator, was markedly stronger in esophagi from Zn-/DFMO+ animals that showed increased apoptosis, whereas increased expression of bcl-2, an inhibitor of apoptosis, was only seen in the highly proliferative, zinc-deficient esophagus (Zn-/DFMO-). At week 12 after NMBA dosing, DFMO reduced the incidence of esophageal tumors from 80 to 4% in zinc-deficient rats. Our data showed that DFMO effectively inhibited the increased esophageal cell proliferation induced by dietary zinc deficiency and reduced the incidence of esophageal tumors induced by a single dose of NMBA in zinc-deficient animals. Our results also indicate a role for increased apoptosis in the mechanism(s) whereby DFMO brings about the inhibition of cell proliferation and tumor induction. These findings support a role for DFMO as a chemopreventive agent. PMID- 9850070 TI - 3-(Iodoacetamido)-benzoylurea: a novel cancericidal tubulin ligand that inhibits microtubule polymerization, phosphorylates bcl-2, and induces apoptosis in tumor cells. AB - 3-(Iodoacetamido)-benzoylurea (3-IAABU) is a newly synthesized antitubulin compound with a molecular weight of 347. 3-IAABU exhibited anticancer activity in a variety of tumor cell lines with ID90 in the range of 0.015-0.29 microM for leukemic cells and 0.06-0.92 microM for solid tumors. Higher selectivity against malignant cells was observed with 3-IAABU than that with vinblastine and paclitaxel. It inhibits microtubule assembly in tubulin systems either with or without microtubule-associated proteins (ID50 was 0.1 microM and 1.2 microM, respectively) and microtubule depolymerization was not affected, indicating an inhibition of polymerization by binding of 3-IAABU to the heterodimeric subunit of tubulin. 3-IAABU was shown to inhibit the binding of colchicine, a subunit binding compound, but did not inhibit binding of vinblastine and guanosine 5' triphosphate/guanosine 5'-diphosphate, indicating that colchicine site corresponds to the site that 3-IAABU locates. Tumor cells treated with 3-IAABU showed scattered chromosomes in metaphase. Normal microtubule architecture or spindle apparatus was absent in these cells; instead, punctuated aggregates of tubulin were found by an immunofluorescent staining. Cell cycle analyses showed an accumulation of tumor cells at M phase after a 4-h treatment with 3-IAABU. The phosphorylated bcl-2 representative of an inactivated form of the oncoprotein was found in the cells 12 h after treatment with 3-IAABU. These cells progressed to apoptosis within 16 h. As a new tubulin ligand, 3-IAABU could be a promising agent in cancer chemotherapy. PMID- 9850072 TI - Attenuation of telomerase activity by a hammerhead ribozyme targeting the template region of telomerase RNA in endometrial carcinoma cells. AB - Telomerase activity is found in almost all carcinoma cells but not in most somatic cells, suggesting that telomerase is an excellent target for cancer therapy. We designed hammerhead ribozymes against human telomerase RNA and studied their possible use as a tool for cancer therapy. Three ribozymes targeting the 3' end of the GUC sequence at 33-35 (the template region), 168-170, and 313-315 from the 5' end of telomerase RNA were designed. In a cell-free system, these three hammerhead ribozymes efficiently cleaved the RNA substrate. When these ribozyme RNAs were introduced into Ishikawa cells, which are endometrial carcinoma cells, only a ribozyme targeting the RNA template region could diminish the telomerase activity. Next we subcloned the ribozyme sequence into an expression vector and introduced this into AN3CA cells, which are endometrial carcinoma cells. The clones that were obtained showed reduced telomerase activity and telomerase RNA with expression of the ribozyme. These data suggest that the ribozyme against the RNA template region is a good tool to repress telomerase activity in cancer cells. PMID- 9850071 TI - Drug resistance patterns of human neuroblastoma cell lines derived from patients at different phases of therapy. AB - To determine whether neuroblastomas acquire a sustained drug-resistant phenotype from exposure to chemotherapeutic agents given to patients in vivo, we studied neuroblastoma cell lines established at different points of therapy: six at diagnosis before therapy (DX), six at progressive disease during induction therapy (PD-Ind), and five at relapse after intensive chemoradiotherapy and bone marrow transplantation (PD-BMT). Cells were maintained in the absence of drug selective pressure. Dose-response curves of melphalan, cisplatin, carboplatin, doxorubicin, and etoposide for the cell line panel were determined by measuring cytotoxicity with a 96-well-plate digital imaging microscopy (DIMSCAN) microassay. Drug resistance of cell lines progressively increased with the intensity of therapy delivered in vivo. The greatest resistance was seen in PD BMT cell lines: IC90 values in PD-BMT cell lines were higher than clinically achievable drug levels by 1-37 times for melphalan, 1-9 times for carboplatin, 25 78 times for cisplatin, 6-719 times for doxorubicin, and 3-52 times for etoposide. Genomic amplification of MYCN did not correlate with resistance. Cross resistance by Pearson correlation (r > or = 0.6) was observed between: (a) cisplatin + doxorubicin; (b) carboplatin + cisplatin, etoposide, or melphalan; (c) etoposide + cisplatin, melphalan, or doxorubicin. These data indicate that during therapy, neuroblastomas can acquire resistance to cytotoxic drugs because of the population expansion of tumor cells possessing stable genetic or epigenetic alterations that confer resistance. PMID- 9850073 TI - (E)-2'-deoxy-2'-(fluoromethylene) cytidine potentiates radioresponse of two human solid tumor xenografts. AB - Antitumor and radiosensitizing effects of (E)-2'-deoxy-2'-(fluoromethylene) cytidine (FMdC), a novel inhibitor of ribonucleotide reductase, were evaluated on nude mice bearing s.c. human C33-A cervix cancer and U-87 MG glioblastoma xenografts. FMdC given once daily has a dose-dependent antitumor effect. The maximum tolerated dose in the mice was reached with 10 daily i.p. administrations of 10 mg/kg over 12 days. In the case of radiotherapy (RT) alone (10 fractions over 12 days), the radiation dose required to produce local tumor control in 50% of the treated C33-A xenografts was 51.0 Gy. When combined with FMdC, the radiation dose required to produce local tumor control was reduced to 41.4 and 38.2 Gy, at respective doses of 5 and 10 mg/kg given i.p. 1 h before each irradiation. The corresponding enhancement ratios (ERs) were 1.2 and 1.3, respectively. In U-87 MG xenografts, when 5-20 mg/kg FMdC combined with 30 or 40 Gy of RT, the combination treatment produced a significantly increased growth delay as compared with RT alone (P < or =0.002). The ERs of 5, 10, and 20 mg/kg FMdC at a dose of 30 Gy were 2.0, 1.4, and 1.8, respectively. At the 40-Gy level, ERs of 10 and 20 mg/kg FMdC were 1.4 and 1.7. When FMdC was combined with 50 Gy of RT, an increased long-term remission rate of 80-88.9% was observed, as compared with 25% for RT alone (P <0.05). FMdC produced moderate myelosuppression in the mice bearing cervix cancer, whereas leukocytosis occurred in the mice bearing glioblastoma at a low dose. Slightly increased skin toxicity (only with U 87 MG tumor) was observed, as compared with RT alone. In conclusion, FMdC is a potent cytotoxic agent and able to modify the radiation response of C33-A and U 87 MG xenografts. PMID- 9850074 TI - Expression, characterization, and detection of human uridine phosphorylase and identification of variant uridine phosphorolytic activity in selected human tumors. AB - Uridine phosphorylase (UPase) catalyzes the reversible phosphorolysis of uridine to uracil. We purified the enzyme from the murine colon 26 tumor using a two-step procedure through 5-amino-benzylacyclouridine affinity chromatography. Antibodies raised in rabbits against the purified protein revealed single bands in Western blots of normal human tissue and tumor extracts. The polyclonal antibody used to screen a human liver expression library allowed the isolation of a 1.2-kb clone that contained the entire open reading frame of the human UPase. The UPase cDNA has been expressed as a fusion protein in Escherichia coli using the pMal-C2 vector. The kinetic analysis demonstrated that the recombinant UPase preferentially uses uridine, 5-fluorouracil, and uracil as substrates, although lower levels of activity were observed with 2-deoxyuridine and thymidine. Clinical samples of human tumors and adjacent normal tissues were assayed for phosphorolytic activity and sensitivity to 5-benzylacyclouridine (BAU), a potent inhibitor of the enzyme presently in Phase I-II clinical trial. Activity in normal tissues appeared to be low but very sensitive to BAU (approximately 90% inhibition at 10 microM). Tumors had generally 2-3-fold greater activity compared with adjacent normal tissues. In breast cancer specimens and head-neck squamous carcinomas, however, uridine cleavage was only partially inhibited (40-60%) by 10 or 100 microM BAU. The BAU-insensitive activity requires phosphate and pH conditions similar to the normal enzyme, and the new phosphorolytic activity was independent from thymidine phosphorylase. The BAU-insensitive phosphorolytic activity in selected tumors, coupled with the potent inhibitory activity of BAU against the "classical" uridine phosphorylase in normal human tissues, provides the rationale for combining BAU with 5-fluorouracil in the treatment of breast and head-neck tumors. PMID- 9850075 TI - Photodynamic therapy of orthotopic prostate cancer with benzoporphyrin derivative: local control and distant metastasis. AB - This is the first report of photodynamic therapy (PDT) in an orthotopic prostate tumor model and shows that PDT combined with surgery (tumor bed sterilization) gave significant local control of the primary tumor and significant reduction in distant metastases. By contrast, either treatment alone (surgery or PDT) gave relatively poorer local control, and PDT gave a significant increase in the mean number of lung metastases. The MatLyLu variant of the Dunning 3327 rat prostate cancer cell line, which has been selected to be metastatic to lymph nodes and lungs, was injected into the ventral lobe of the rat prostate. After 7 days, tumors were either treated by surgical removal of the ventral lobe, PDT with liposomal benzoporphyrin derivative monoacid ring A, or a combination of surgery, followed by PDT of the tumor bed. Results after 21 days showed a reduction in prostate tumor weight in all groups compared with controls, which became highly significant only for the combination group (17% of control mean tumor weight; P < 0.001; 7 of 13 clinical complete responses). The combination treatment also led to a reduction in lymph node metastasis and reductions in both the frequency and mean number of lung metastases compared with other treatment groups. The PDT alone group, however, had a mean number of lung metastases per animal, which was nine times the control group and 34 times the combination group. These findings suggest that a tumor bed sterilization approach may be promising for locally advanced prostate cancer and suggest that factors other than local control may need to be evaluated when considering PDT for primary prostate cancer. PMID- 9850076 TI - Inhibition of insulin-like growth factor I receptor expression in neuroblastoma cells induces the regression of established tumors in mice. AB - Several lines of evidence now indicate that type 1 insulin-like growth factor receptor (IGF1R) function may be particularly important in the pathogenesis of the pediatric cancer neuroblastoma. Modulating the expression of specific genes involved in neuroblastoma tumorigenesis could provide a much needed alternative treatment strategy for poor prognosis disease. We now report construction of an antisense expression vector to the IGF1R that markedly reduces cellular IGF1R levels and inhibits the proliferation and clonogenicity of neuroblastoma cells in vitro but not that of IGF1R null cells. This antitumor activity is associated with the induction of apoptotic cell death in transfected cells, as measured by annexin V staining and flow cytometry. Direct injection of this vector into established tumors growing in syngeneic mice results in a marked inhibition of tumor growth with complete and durable tumor regression in one-half of the animals. This effect appears to be immunologically mediated in that vector injection of neuroblastoma tumors growing in severe combined immunodeficiency mice results in only modest delay of tumor growth. Our results suggest that inhibition of IGF1R expression by direct intratumoral delivery of an antisense construct could provide a novel therapeutic approach in the management of poor prognosis neuroblastoma. PMID- 9850077 TI - EblacZ tumor dormancy in bone marrow and lymph nodes: active control of proliferating tumor cells by CD8+ immune T cells. AB - A well-defined lacZ gene tagged DBA/2 lymphoma (EblacZ) was used to examine the role of host immune responses in controlling tumor dissemination and persistence, as well as metastasis. In s.c. and intra-ear pinna-inoculated mice, low numbers of EblacZ cells homed to the bone marrow and lymph nodes. The frequency of bone marrow-residing tumor cells did not change with the growth of primary tumor or with multiple inoculations of tumor cells. The bone marrow-residing tumor cells expressed the proliferation-associated Ki67 antigen and expanded upon CD8+ depletion. In contrast, inoculation of nu/nu or severe combined immunodeficiency mice or of immune-suppressed DBA/2 mice led to the rapid outgrowth of EblacZ cells in the bone marrow and their metastasis to other organs. Transfer of bone marrow from EblacZ immunized MHC congenic or syngeneic DBA/2 donors, but not from naive donors, protected s.c.-inoculated DBA/2 mice. Protection was abrogated by in vitro depletion of CD8+ T cells prior to transfer of bone marrow. These experiments show that bone marrow and lymph nodes are privileged sites where potentially lethal tumor cells are controlled in a dormant state by the immune system. Metastasis may be a consequence of the breakdown of this immune control. PMID- 9850078 TI - Immunity to p53 induced by an idiotypic network of anti-p53 antibodies: generation of sequence-specific anti-DNA antibodies and protection from tumor metastasis. AB - The general overexpression of p53 by different types of tumor cells suggests that p53 immunity might be generally useful for tumor immunotherapy. We describe here the induction of immunity to p53 and resistance to tumor metastasis using an idiotypic network. Mice were immunized with domain-specific anti-p53 monoclonal antibodies (Ab1): PAb-248 directed to the N-terminus; PAb-246 directed to the specific DNA-binding region; or PAb-240 directed to a mutant p53 that does not bind specific DNA. Immunized mice responded by making anti-idiotypic antibodies (Ab2) specific for the Ab1 inducer. Ab1 PAb-246 induced Ab2 that, like p53 itself, could bind the specific DNA oligonucleotide sequence of the p53 responsive element. Mice immunized with Ab1 PAb-240 or PAb-246 spontaneously made Ab3 anti-p53 antibodies that reflected the specificity of their Ab1 inducers: Ab1 PAb-246 induced Ab3 specific for wild-type p53; PAb-240 induced Ab3 specific for mutant p53. Ab1 PAb-248 induced only Ab2. The spontaneously arising Ab3 were of T cell-dependent IgG isotypes. Peptides from the complementarity determining regions of the Ab1 antibodies PAb-240 and PAb-246 could also induce Ab3 anti-p53. Finally, mice that produced Ab3 anti-p53 acquired resistance to tumor metastases. Therefore, an anti-idiotypic network built around certain domains of p53 seems to be programmed within the immune system, specific Ab2 antibodies can mimic the DNA binding domain of p53, and Ab3 network immunity to p53 can be associated with resistance to tumor cells. PMID- 9850079 TI - The relationships between p53-dependent apoptosis, inhibition of proliferation, and 5-fluorouracil-induced histopathology in murine intestinal epithelia. AB - The relationship between acute (<36 h) induction of apoptosis and longer-term (>72 h) intestinal histopathology was systematically investigated in vivo using p53 wild-type (+/+) and null (-/-) mice. Administration of the enterotoxin 5 fluorouracil (5-FU) at either 40 or 400 mg/kg to BDF1 mice induced an acute p53 dependent apoptosis in the crypts of both small intestine and midcolon. Although the amount of apoptosis was of the same order of magnitude at its peak (24 h) at both doses, only 400 mg/kg 5-FU brought about histopathological changes to the gut after 96 h, quantified as losses of crypt and villus cellularity. Only after the administration of 400 mg/kg 5-FU were mitotic index and DNA synthesis significantly suppressed in both small intestinal and midcolonic crypts at 24 h. This correlated with a prolonged, p53-dependent expression of p21waf-1/cip1. In p53 null (-/-) mice, significant reductions in both 5-FU-induced apoptosis and inhibition of cell cycle progression allowed retention of crypt integrity 96 h after 5-FU. These results show that quantitative measures of acute apoptosis in vivo may not accurately predict subsequent pathological changes in the gut. Rather, p53-dependent inhibition of cell cycle progression, together with cell loss by apoptosis, caused a loss of crypt integrity. Importantly, the tissue toxicity of 5-FU was genetically determined at a locus (p53) separate from that directly associated with drug action. PMID- 9850080 TI - Expression of AP-2 transcription factors in human breast cancer correlates with the regulation of multiple growth factor signalling pathways. AB - The AP-2 transcription factors are required for normal growth and morphogenesis during mammalian development. Previous in vitro studies have also indicated that the AP-2 family of proteins may be involved in the etiology of human breast cancer. The AP-2 genes are expressed in many human breast cancer cell lines, and critical AP-2-binding sites are present in both the ERBB-2 (HER2/neu) and estrogen receptor promoters. We have now characterized immunological reagents that enable specific AP-2 family members, including AP-2alpha and AP-2gamma, to be detected in human breast cancer epithelium. Data obtained with these reagents demonstrate that whereas AP-2alpha and AP-2gamma are both present in benign breast epithelia, there is a significant up-regulation of AP-2gamma expression in breast cancer specimens (P = 0.01). There was also a significant correlation between the presence of the AP-2alpha protein and estrogen receptor expression (P = 0.018) and between specimens containing both AP-2alpha/AP-2gamma proteins and ERBB-2 expression (P = 0.003). Furthermore, we detected an association (P = 0.04) between the expression of AP-2gamma and the presence of an additional signal transduction molecule implicated in breast cancer, the insulin-like growth factor I receptor. Analysis of the proximal promoter of the insulin-like growth factor I receptor revealed a novel AP-2-binding site. Thus, AP-2 proteins may directly regulate the transcription of this growth factor receptor. Taken together, these data strongly support a role for the AP-2 gene family in the control of cell growth and differentiation in breast cancer. PMID- 9850081 TI - Reduced COX-2 protein in colorectal cancer with defective mismatch repair. AB - Most colorectal adenomas and carcinomas arise in the setting of chromosomal instability characterized by progressive loss of heterozygosity. In contrast, approximately 15-20% of colorectal neoplasms arise through a distinct genetic pathway characterized by microsatellite instability (MSI) associated with frequent loss of expression of one of the DNA mismatch repair enzymes, most often hMLH1 or hMSH2. These distinct genetic pathways are reflected by differences in tumor histopathology, distribution in the colon, prognosis, and dwell time required for progression from adenoma to carcinoma. To determine whether these two groups of tumors differ in their expression of cyclooxygenase-2 (COX-2), a putative chemopreventative target, immunostaining for this protein was performed in colorectal cancers categorized by the presence (n = 41) and absence (n = 66) of defective mismatch repair. Defective mismatch repair was defined by the presence of tumor microsatellite instability (MSI-H, > or =40% of markers demonstrating instability) and by the absence of protein expression for either hMLH1 or hMSH2. Overall, our results showed that low or absent COX-2 staining was significantly more common among tumors with defective mismatch repair (P = 0.001). Other features predictive of low COX-2 staining included marked tumor infiltrating lymphocytosis, and solid/cribiform or signet ring histological patterns. These observations indicate that colorectal cancers with molecular and phenotypic characteristics of defective DNA mismatch repair express lower levels of COX-2. The clinical implications of this biological distinction remain unknown but should be considered when assessing the efficacy of COX-2 inhibitors for chemoprevention in patients whose tumors may arise in the setting of defective DNA mismatch repair. PMID- 9850082 TI - Clinicopathological significance of Fhit protein expression in stage I non-small cell lung carcinoma. AB - Abnormalities in structure and expression of the FHIT gene have been detected in a considerable fraction of primary lung tumors. Previous reports indicated that FHIT gene alterations can be simply detected by immunohistochemical methods. Therefore, we investigated the association of Fhit expression with clinicopathological features and allelic imbalance (AI) at the FHIT locus in 105 stage I non-small cell lung cancers (NSCLC) by the immunohistological method and PCR analysis. Thirty-six of 105 (34%) tumors showed marked reduction of Fhit immunoreactivity. Fhit expression was markedly reduced in most squamous cell carcinomas (24 of 28, 86%), whereas such a reduction was detected only in a small subset of adenocarcinomas (7 of 67, 10%; P < 0.001). A marked reduction of Fhit protein expression was observed more frequently in patients with a smoking history (32 of 80, 40%) than in patients without a smoking history (4 of 25, 16%; P = 0.013). These results indicate that FHIT gene alterations preferentially occur in squamous cell carcinomas and in smokers. Furthermore, a reduction of Fhit protein expression in tumor cells was associated with a poorer survival of patients with stage I NSCLC, irrespective of histological subtypes of tumors (P = 0.005; log-rank test). Fhit expression was reduced preferentially in tumors with AI at the FHIT locus; however, AI at the FHIT locus did not correlate with patients' survival (P = 0.262; log-rank test). These results suggested that Fhit protein expression could be a useful molecular marker for the prognosis of patients with surgically resected stage I NSCLC. PMID- 9850083 TI - Nuclear factor kappaB dominant negative genetic constructs inhibit X-ray induction of cell adhesion molecules in the vascular endothelium. AB - X-ray-induced expression of inflammatory mediators has been proposed to contribute to radiation injury in normal tissues. Radiation-inducible inflammatory mediators include the cell adhesion molecule (CAM) E-selectin and the intercellular adhesion molecule (ICAM)-1. Nuclear factor (NF)kappaB is activated by X-rays and may participate in the transcriptional regulation of each of these inflammatory mediators. To determine whether NFkappaB inhibition abrogates X-ray induction of inflammatory mediators, we used two experimental approaches including NFkappaB inhibitory drugs and a dominant negative genetic construct. Human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells were treated with the NFkappaB inhibitors ALLN, PDTC, NAC, and MG132. After irradiation, E-selectin or ICAM-1 was measured by fluorescence activated cell-sorting analysis. E-selectin and ICAM-1 expression was measured by use of immunofluorescence and fluorescence-activated cell-sorting analysis. E selectin expression increased 7-fold, and ICAM-1 expression increased 4-fold after irradiation. All of the inhibitors attenuated E-selectin expression after irradiation. ALLN and MG132 attenuated radiation-induced ICAM expression. However, PDTC and NAC induced increased expression of ICAM-1 in HUVECs. Inhibition of X-ray induction of ICAM by these agents could not be demonstrated. In separate experiments, the NFkappaB dominant negative genetic construct was cotransfected with the promoter-reporter constructs by means of Lipofectin reagent. The ICAM promoter-reporter construct consists of the 1.2-kb segment of the human ICAM promoter upstream of the transcriptional start site linked to the luciferase reporter gene (pGL.FL-Luc). The E-selectin promoter-reporter construct consists of 525 bp upstream of the transcriptional start site of the human E selectin promoter linked to the human growth hormone reporter gene (pE525-GH). Endothelial cells transfected with the ICAM-1 promoter-reporter construct showed a 3-fold induction after irradiation. Likewise, cells transfected with pE525-GH showed a 7-fold induction after irradiation. When cotransfected with the CAM reporter-promoter constructs, the NFkappaB dominant negative genetic construct abolished X-ray-induced transcriptional activation of the E-selectin and ICAM-1 promoters. NFkappaB inhibition is, therefore, a means of abrogating radiation induced expression of CAMs. PMID- 9850084 TI - Aging and DNA methylation in colorectal mucosa and cancer. AB - DNA methylation of promoter-associated CpG islands may function as an alternate mechanism of silencing tumor suppressor genes in multiple neoplasias including colorectal cancer. De novo methylation of genes appears to be an early and frequent event in most neoplasias. For the ER and IGF2 genes, we have previously shown that methylation actually begins in the normal colon mucosa as an age related event and progresses to hypermethylation in cancer. In this study, we have determined the frequency of age-related methylation in normal colonic mucosa among the genes hypermethylated in colorectal cancer. We studied six genes, including N33, MYOD, p16, HIC-1, THBS1, and CALCA. The N33 gene showed partial methylation in normal colon mucosa, which was age-related (r = 0.7; P = 0.003 using regression analysis). Adenomas and cancers showed further hypermethylation at this locus. Similarly, the MYOD gene showed age-related methylation in normal colon mucosa (r = 0.7; P < 0.00001 using regression analysis) and hypermethylation in cancers. Age-related methylation seems to be gene specific, because p16, THBS1, HIC-1, and CALCA were not affected. Furthermore, this process may also be modulated by tissue-specific factors. Our study suggests that aging is a major contributing factor to hypermethylation in cancer. PMID- 9850085 TI - Heat shock protein 27 enhances the tumorigenicity of immunogenic rat colon carcinoma cell clones. AB - The REG and PRO cell clones were obtained from a colon adenocarcinoma induced in a BDIX rat by 1,2-dimethylhydrazine. When injected s.c. into syngeneic hosts, REG cells induce tumors that regress in less than 3 weeks, whereas PRO cells, like parental cells, induce progressive tumors. Here, we show that compared to PRO cells, REG cells are more sensitive to cell death induced by anticancer drugs. The small heat shock protein (HSP) 27 is not expressed or inducible in REG clones, whereas it is abundantly expressed and inducible by heat shock in PRO clones. The expression of HSP27 in REG cells increases their resistance to apoptosis in vitro and dramatically enhances their tumorigenicity when injected s.c. into syngeneic rats. HSP27 expression in REG cells both increases tumor size and delays tumor regression. This increased tumorigenicity is associated with a substantial decrease of in vivo tumor cell apoptosis. We conclude that HSP27 expression in malignant cells increases their tumorigenicity in syngeneic animals. In combination with the role of HSP27 in tumor cell resistance to cytotoxic agents, its contribution to tumorigenicity makes this protein a potential target for antitumoral therapy. PMID- 9850086 TI - The matrix metalloproteinase matrilysin influences early-stage mammary tumorigenesis. AB - Overexpression of the epithelial specific matrix metalloproteinase matrilysin (MAT) has been correlated with enhanced tumorigenicity and tumor cell invasion using in vitro model systems. We have determined the effects of MAT expression on the development of mammary tumorigenesis using transgenic mice that express human MAT under the control of the mouse mammary tumor virus (MMTV)-long terminal repeat promoter/enhancer. Examination of mammary glands from multiparous MMTV-MAT animals revealed the development of premalignant hyperplastic alveolar nodules in 50% of aged females. MMTV-MAT mice were mated with MMTV-neu transgenic mice to determine the effect of MAT on neu-induced mammary tumorigenesis. Bigenic MMTV MAT/neu female offspring developed primary mammary tumors approximately 13 weeks earlier than did MMTV-neu controls. The mechanism of enhanced neu-induced tumorigenesis was explored. No discernible difference in Neu receptor dimerization or activation was detected in MMTV-MAT/neu tumors or mammary glands compared to MMTV-neu controls. A similar percentage of MMTV-MAT/neu and MMTV-neu tumors acquired deletions in the neu transgene, which have previously been shown to result in constitutive receptor activation. The presence of premalignant nodules and the accelerated development of oncogene-induced mammary tumors suggest that expression of MAT in the mammary epithelium contributes to early stage mammary tumorigenesis. PMID- 9850087 TI - Abundance of heat shock proteins (hsp89, hsp60, and hsp27) in malignant cells of Hodgkin's disease. AB - Heat shock proteins (HSPs) or stress proteins are synthesized by cells in response to environmental stress. Expression of HSPs by cells may have important physiological or pathological implications. In this study, we carried out an immunohistochemical and biochemical examination of low (hsp27), intermediate (hsp60), and high (hsp89) molecular weight HSP expression in reactive lymph nodes and in lymph nodes of patients with various types of lymphomas. In normal or reactive lymphoid tissues, hsp89 is abundant in large "transformed" lymphoid cells and immunoblasts. Hsp60 is widely distributed in lymphoid tissues, whereas hsp27 is absent in all lymphoid cells and histiocytes. Among lymphomas, the Hodgkin's Reed-Sternberg (H-RS) cells in Hodgkin's disease (HD) had the greatest abundance of hsp89 and hsp60 and, in 20% of cases, hsp27, in contrast to a much weaker staining of anti-hsp89 and -hsp60 in the background reactive lymphoid cells. The large lymphoid cells in small lymphocytic lymphoma are also rich in hsp89, but not hsp60 and hsp27. In contrast, the malignant cells in anaplastic large cell lymphoma and most high-grade tumors, including immunoblastic lymphomas, expressed minimal amounts of hsp89 and hsp60 and virtually no hsp27. Thus, the cellular level of HSPs was neither correlated with the proliferative capacity nor with the aggressiveness of the lymphomas. Hsp89, hsp60, and hsp27, as well, serve critical roles in the chaperoning of cellular proteins (e.g., a Mr 43,000 protein) in H-RS cells. The known interactions of HSPs with Rb, p53, peptide-MHC class II complexes, and cofactors of the glucocorticoid hormone receptor have further broadened the importance of HSPs in cell metabolism and in response to extracellular signals for proliferation, differentiation, or growth suppression (or apoptosis) of H-RS cells. Abundant HSP expression is seen only in HD, but not in other lymphomas. Such expression could have vital roles in the pathogenesis of HD. PMID- 9850088 TI - Generation of hepatocytes from oval cell precursors in culture. AB - Although there is experimental evidence supporting the involvement of hepatic stem cells in the pathogenesis of liver cancers, the detection and isolation of these cells remains elusive. A logical approach to detecting these cells would take advantage of their ability to differentiate (or to give rise to cells that differentiate) into hepatocytes. This approach requires an assay system that is conducive to hepatocytic differentiation. Here, we report the development of an in vitro system consisting of a three-dimensional collagen gel matrix and a fibroblast feeder layer that supports hepatocytic differentiation from precursor epithelial (oval) cell lines. The LE/2 and LE/6 oval cell lines used in this study are nontumorigenic cells that are derived from the livers of adult rats fed a choline-deficient diet containing 0.1% ethionine for 2 and 6 weeks, respectively. These lines consist of small cells that are phenotypically immature with few cytoplasmic organelles and a high nuclear-to-cytoplasmic ratio. After 4 weeks in the three-dimensional culture system, these cells acquired typical hepatocytic morphology. By electron microscopy, the cells formed canalicular structures that are typical of hepatocytes and were organelle rich, displaying peroxisomes, abundant mitochondria, and rough endoplasmic reticulum. The cells produced albumin and displayed a cytokeratin (CK) pattern typical of hepatocytes (CK 8 and CK 18-positive and CK 19-negative). The presence of a mesenchymal cell feeder layer was essential for supporting hepatocytic differentiation. Without a feeder layer but in the presence of hepatocyte growth factor and/or keratinocyte growth factor, the precursor cells formed ductal structures, suggestive of differentiation along the bile duct lineage. The three-dimensional system described provides direct proof of the lineage generation capacity of oval cells. It offers a model to study factors that may be important for hepatocytic differentiation from precursor cells and a means to assay cell populations for their ability to give rise to normal and transformed hepatocytes. PMID- 9850089 TI - The RET/PTC3 oncogene: metastatic solid-type papillary carcinomas in murine thyroids. AB - Our research goal is to better understand the mechanisms controlling the initiation and progression of thyroid diseases. One such disease, papillary thyroid carcinoma (PTC), is the leading endocrine malignancy in the United States. Recently, a family of related fusion proteins, RET/PTC1-5, has been implicated in the early stages of PTC. Although all five members of this family have the c-RET proto-oncogene kinase domain in their COOH terminus, little is known about how these genes alter follicular cell biology. Consequently, to answer questions related to the mechanism of the RET/PTC fusion protein action, we have devised a molecular genetic strategy to study PTC using a mouse model of thyroid disease. A new member of this fusion oncogene family, RET/PTC3, which has been implicated in more cases of solid tumor carcinoma (79%) than PTC1 or PTC2 and predominates (80%) in radiation-induced thyroid cancer of children, was investigated in our study. We have generated transgenic mice expressing human RET/PTC3 exclusively in the thyroid. These mice develop thyroid hyperplasia, solid tumor variants of papillary carcinoma and metastatic cancer. This new transgenic line will be useful in deciphering the molecular and biological mechanisms that cause PTC and histological variations in humans. PMID- 9850090 TI - SPARC/osteonectin induces matrix metalloproteinase 2 activation in human breast cancer cell lines. AB - Activation of the matrix metalloproteinase 2 (MMP-2) has been shown to play a major role in the proteolysis of extracellular matrix (ECM) associated with tumor invasion. Although the precise mechanism of this activation remains elusive, levels of the membrane type 1-MMP (MT1-MMP) at the cell surface and of the tissue inhibitor of MMP-2 (TIMP-2) appear to be two important determinants. Induction of MMP-2 activation in cells cultivated on collagen type I gels indicated that the ECM is important in the regulation of this process. In this study, we show that SPARC/osteonectin, a small ECM-associated matricellular glycoprotein, can induce MMP-2 activation in two invasive breast cancer cell lines (MDA-MB-231 and BT549) but not in a noninvasive counterpart (MCF-7), which lacks MT1-MMP. Using a set of peptides from different regions of SPARC, we found that peptide 1.1 (corresponding to the NH2-terminal region of the protein) contained the activity that induced MMP-2 activation. Despite the requirement for MT1-MMP, seen in MCF-7 cells transfected with MT1-MMP, the activation of MMP-2 by SPARC peptide 1.1 was not associated with increased steady-state levels of MT1-MMP mRNA or protein in either MT1-MMP-transfected MCF-7 cells or constitutively expressing MDA-MB-231 and BT549 cells. We did, however, detect decreased levels of TIMP-2 protein in the media of cells incubated with peptide 1.1 or recombinant SPARC; thus, the induction of MMP-2 activation by SPARC might be due in part to a diminution of TIMP-2 protein. We conclude that SPARC, and specifically its NH2-terminal domain, regulates the activation of MMP-2 at the cell surface and is therefore likely to contribute to the proteolytic pathways associated with tumor invasion. PMID- 9850091 TI - Association of the decreased expression of alpha3beta1 integrin with the altered cell: environmental interactions and enhanced soft agar cloning ability of c-myc overexpressing small cell lung cancer cells. AB - Small cell lung cancer (SCLC) is a highly invasive and metastatic tumor, and the decreased expression of alpha3beta1 integrin may contribute to its virulence. Alpha3beta1 is a critical integrin for pulmonary development and epithelial integrity, and its reduced expression has been linked to the increased malignancy and invasion of other cancers. The amplification of the c-myc oncogene is seen frequently in relapsed SCLC tumors and is associated with a worsened prognosis. In the present study using a model of SCLC tumor progression, overexpression of c myc in a classic SCLC cell line, NCI H209, enhanced in vitro features of tumorigenesis, altered the relationships between cell and environment, and markedly down-regulated the expression of the alpha3 integrin subunit at both the transcript and protein levels. This inverse relationship between the expression of the alpha3 integrin subunit and c-myc is mimicked by other c-myc overexpressing SCLC cell lines. Restoring alpha3 expression in the myc transfected 209 cells reversed the effects of c-myc: alpha3 transfection increased cell:cell adhesion and reduced soft agar cloning without affecting the in vitro doubling time. The diminished soft agar cloning produced by alpha3 transfection was reversed by an antibody that specifically engages alpha3beta1 integrins, P1B5. These results suggest first, that alpha3beta1 integrin mediates homotypic adhesion of SCLC cells, and second, that unengaged alpha3beta1 integrin suppresses the growth of disaggregated SCLC cells. Thus, the down-regulation of the alpha3 integrin subunit may contribute to the enhanced tumorigenicity of c myc-overexpressing SCLCs by allowing the growth of tumor cells that have reduced contact with ligand-expressing substratum or cells, a condition that occurs during the growth of the primary tumor, tumor invasion, and metastasis. PMID- 9850092 TI - Mucin gene expression in ovarian cancers. AB - Ovarian cancer is a highly lethal disease with metastases present in the majority of patients at the time of diagnosis. The molecular mechanisms underlying the metastatic process of this cancer are not well understood. One family of cell associated and secreted glycoproteins, the mucin glycoproteins, has been implicated in events leading to metastasis of several epithelial cancers including gastrointestinal and lung cancers. The purpose of this study was to characterize mucin gene expression in ovarian cancers and relate expression to tumor histology, stage, and patient survival. RNA was isolated from 29 epithelial ovarian cancers, 1 neuroendocrine carcinoma, 3 mixed mesodermal tumors, and two transformed, yet nonmalignant, ovarian epithelial cell lines. The expression of mucin genes, MUC1, 2, 3, 4, 5AC and 5B, was determined by northern analyses. Epithelial ovarian cancers expressed several mucins including MUC1, 2, 4, and 5AC; MUC3 and 5B were rarely expressed. In contrast, the transformed nonmalignant ovarian epithelial cell lines expressed only MUC1 and 5AC. Although there was no correlation of mucin expression with tumor histology, there was a significant decrease in expression of MUC3 and MUC4 with increasing cancer stage (P < 0.05). In addition, a trend toward improved patient survival occurred with increased expression of MUC4. These observations suggest a relationship between mucin gene expression and the metastatic process in epithelial ovarian cancers. Additional investigation of MUC3 and MUC4 in ovarian cancers may lead to new approaches for early detection and therapy. PMID- 9850093 TI - Retinoic acid and interferon alpha act synergistically as antiangiogenic and antitumor agents against human head and neck squamous cell carcinoma. AB - Head and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy in which multiple independent lesions develop over time throughout the mucosa of the upper aerodigestive tract. Therefore, the comprehensive treatment of this neoplasm must include a chemopreventive arm to hold premalignant lesions in check, a role well-suited to antiangiogenic agents. Retinoic acid (RA) and interferon alpha (IFN-alpha), drugs with known biological activity against HNSCC when used individually, are also inhibitors of angiogenesis. Here we show that they are remarkably synergistic antiangiogenic agents able to inhibit both the growth and the neovascularization of HNSCC injected into the floor of the mouth of nude mice. The mechanism of action of these drugs as antiangiogenic agents was 2-fold. They decreased the angiogenic activity of the tumor cells, and they caused the endothelial cells to become refractory to inducers of angiogenesis. When tumor cells were treated in vitro with IFN-alpha A/D, there was a dramatic drop in their secretion of interleukin-8, the major angiogenic factor produced by these tumors. When combined with RA, which causes tumor cells to secrete an inhibitor of angiogenesis, there was a synergistic inhibition of both tumor cell growth and secreted angiogenic activity. The combination of RA and IFN-alpha also acted synergistically on endothelial cells by reducing their responsiveness to both interleukin-8 and tumor conditioned media. Doses of each drug could be reduced by two logs without loss of activity. When animals bearing human HNSCC tumor cells were treated systemically with a combination of RA and IFN-alpha A/D at doses that were ineffective when used alone, dramatic decreases in both tumor growth and tumor angiogenesis were seen. These data suggest that the use of antiangiogenic mixtures may be a particularly effective way to design future chemoprevention protocols against HNSCC. PMID- 9850094 TI - Prognostic value of beta1,6-branched oligosaccharides in human colorectal carcinoma. AB - Increase of beta1,6-branched oligosaccharides is possibly associated with tumor progression and lymph node metastasis. The aim of this study was to determine the prognostic value of beta1,6 branches in human colorectal carcinoma. Expression of beta1,6 branches was histochemically evaluated using the leukoagglutinating Phaseolus vulgaris lectin, PHA-L, in 92 clinically documented colorectal carcinomas, of which 31 had formed lymph node metastases. The follow-up time ranged between 4 and 14 years (median, 10.3 years). A PHA-L staining index (SI), taking into account staining intensity and its percentage of tumor cut surface area, was established. The carcinoma SI was highly associated with the disease free survival (P = 0.004) and overall survival (P = 0.005). Patients with a carcinoma SI of >1, as compared to those with a SI of < or =1, were at significantly higher risk for tumor recurrence, with a shorter disease-free survival (hazard ratio = 2.59, P = 0.005) and significant higher risk of death with shorter overall survival (hazard ratio = 2.51, P = 0.007). The carcinoma SI was also associated with the presence of lymph node metastases. We conclude that PHA-L staining in human colorectal carcinoma sections provides an independent prognostic indicator for tumor recurrence and patient survival and is associated with the presence of lymph node metastases. PMID- 9850096 TI - Bcl-2-mediated resistance to apoptosis is associated with glutathione-induced inhibition of AP24 activation of nuclear DNA fragmentation. AB - Studies on the mechanism of apoptosis in this laboratory support a model in which signal transduction involving caspase 3 leads to activation of a serine protease called Mr 24,000 apoptotic protease (AP24), which then induces internucleosomal DNA fragmentation in the nucleus. This study examined the effect of Bcl-2 overexpression on activation of AP24 and the induction of DNA fragmentation by AP24 in isolated nuclei. It was demonstrated that overexpression of Bcl-2 in either HL-60 or PW leukemia cell lines suppressed activation of AP24 induced by either tumor necrosis factor or UV light and protected cells from apoptosis. Furthermore, nuclei isolated from Bcl-2-overexpressing cells were relatively resistant to internucleosomal DNA fragmentation induced by AP24 isolated from apoptotic cells. Bcl-2-overexpressing cells that were nutritionally depleted of glutathione (GSH) became sensitive to tumor necrosis factor- or UV light-induced activation of AP24 and underwent apoptotic cell death. Moreover, nuclei isolated from Bcl-2-overexpressing cells that were depleted of GSH became sensitive to AP24-induced DNA fragmentation. The addition of exogenous GSH blocked the proteolytic activity of AP24, as well as its ability to induce DNA fragmentation in normal isolated nuclei. These results indicate that Bcl-2 can attenuate at least two events in the AP24 apoptotic pathway: activation of AP24 and induction of DNA fragmentation by activated AP24. Furthermore, agents that deplete intracellular levels of GSH may have therapeutic use in the sensitization of Bcl 2-overexpressing cancer cells to apoptotic cell death. PMID- 9850095 TI - Vascular endothelial growth factor inhibits apoptotic death in hematopoietic cells after exposure to chemotherapeutic drugs by inducing MCL1 acting as an antiapoptotic factor. AB - We reported previously that vascular endothelial growth factor (VEGF) inhibits the apoptotic death of hematopoietic cells that is induced by exposure to ionizing radiation (O. Katoh et al., Cancer Res., 55: 5687-5692, 1995). In this study, we show that VEGF also inhibits apoptotic cell death that is induced by exposure to the chemotherapeutic drugs etoposide and doxorubicin. To elucidate the molecular mechanisms underlying this inhibitory effect of VEGF, we examined expression levels of BCL2 family proteins in CMK86, a human leukemia cell line, after treatment with VEGF. Northern blotting and immunoblotting analyses revealed that the expression level of MCL1, a member of the BCL2 family, was increased by VEGF. Moreover, to examine the effects of MCL1 on apoptotic cell death induced by exposure to etoposide, we generated a clonal U937 myeloid leukemia cell line transfected with vectors that promoted the constitutive expression of MCL1. MCL1 decreased the caspase 3 activity induced by exposure to etoposide and increased the viability of the transfected cells after etoposide exposure. Therefore, MCL1 may be involved in the inhibitory effect of VEGF on apoptotic cell death. PMID- 9850097 TI - MUC1 synthetic peptide inhibition of intercellular adhesion molecule-1 and MUC1 binding requires six tandem repeats. AB - We reported recently that breast cancer-associated MUC1 is a ligand for intercellular adhesion molecule-1 (ICAM-1; L. H. Regimbald et al., Cancer Res., 56: 4244-4249, 1996). We report here the results of a competitive indirect binding assay to detect the molecular requirements for binding between ICAM-1 and MUC1. The assay involved inhibition of the binding of recombinant human ICAM-1 to a murine breast adenocarcinoma cell line transfected with human MUC1. The addition of a library of human MUC1 synthetic peptides ranging from 9 to 24 amino acids (aa) showed minimal or no inhibition. However, a 120-aa peptide that corresponds to six tandem repeats of the human mucin MUC1 was as effective an inhibitor as purified tumor MUC1 and MUC1 epitope (PDTRPAP)-specific antibody (B27.29). We conclude that the number of MUC1 tandem repeats necessary for an ordered tertiary structure (D. Fontenot et al., Cancer Res., 53: 5386-5394, 1993) is also important for ICAM-1 recognition. These findings are similar to those described recently for MUC1 induction of T-cell anergy (B. Agrawal et al., Nat. Med., 4: 43-49, 1998). This suggests that the anergy induction by MUC1 may be due to ICAM-1 binding by MUC1. PMID- 9850098 TI - MUC1 is a novel marker for the type II pneumocyte lineage during lung carcinogenesis. AB - Abnormalities in mucin-type glycoprotein expression have been documented in a variety of cancers, identifying these molecules as targets for immunologically based therapies and prognostic/diagnostic assays. We examined the expression of the membrane-bound MUC1 mucin in normal, histologically atypical, and neoplastic lung to determine its potential contribution to lung carcinogenesis. In vivo, intense MUC1 immunoreactivity was present in normal type II pneumocytes as well as in a range of atypical lesions derived from type II cells and >60% of primary and metastatic non-small cell lung cancers. Expression was not associated with altered survival, although it was highly correlated with the adenocarcinoma histology. A carcinogenesis model using 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl) 1-butanone-exposed hamsters revealed that MUC1 mRNA increased prior to the histological appearance of tumors. In vitro studies using MUC1 expressing non small cell lung cancer cell lines revealed that differentiation away from a type II cell lineage was associated with dramatic down-regulation of MUC1. We propose that MUC1 is a powerful new marker for the type II pneumocyte cell lineage that allows us to follow the type II pneumocyte lineage during the process of lung carcinogenesis. PMID- 9850099 TI - Report from THE MARCH Research Task Force. September 25-26, 1998. PMID- 9850100 TI - Should hospital diets meet the dietary guidelines for healthy persons? PMID- 9850101 TI - Should hospital diets meet the dietary guidelines for healthy persons? PMID- 9850102 TI - Should hospital diets meet the dietary guidelines for healthy persons? PMID- 9850103 TI - Should hospital diets meet the dietary guidelines for healthy persons? PMID- 9850104 TI - ADA's campaign for coverage of MNT: success is in sight! American Dietetic Association. Medical nutrition therapy. PMID- 9850105 TI - Grandma eats like a bird: helping caregivers improve the nutrition of older persons at home. PMID- 9850106 TI - An assessment of the scientific basis for attempting to define the Dietary Reference Intake for beta carotene. PMID- 9850107 TI - Dietary Reference Intakes, antioxidants, and beta carotene. PMID- 9850109 TI - Demographic and psychosocial predictors of fruit and vegetable intakes differ: implications for dietary interventions. AB - OBJECTIVE: The National Cancer Institute (Rockville, Md) has launched a nationwide initiative--5 A Day for Better Health--to encourage consumption of fruits and vegetables. Because the tastes and culinary uses of fruits and vegetables differ, however, it is not known whether a general 5-A-Day message is an effective intervention strategy. This study examined whether there are differences between the demographic and psychosocial correlates of fruit and vegetable intakes. DESIGN: Data are from the Washington State Cancer Risk Behavior Survey (1995-1996), a cross-sectional, random-digit-dial telephone survey representative of the adult population of Washington State. SUBJECTS/SETTING: Interviews were completed with 1,450 adults. Data were collected about demographic characteristics, health status, health-related behavior, fruit and vegetable intakes, and the following diet-related psychosocial factors: beliefs, motives, barriers, attitudes, and stages of dietary change. STATISTICAL ANALYSES: Multivariate linear regression analysis was used to test whether the associations of demographic characteristics and psychosocial factors with fruit intake differed from associations with vegetable intake. RESULTS: In general, health status, health-related behavior, and psychosocial factors were more strongly associated with fruit intakes than vegetable intakes. For example, regular exercisers consumed 0.44 more daily servings of fruits and 0.36 more servings of vegetables than nonexercisers. Compared with those in the preaction stage of dietary change, adults in the maintenance stage consumed 0.99 more daily servings of fruits and 0.68 more servings of vegetables. Intrinsic motivations for eating a healthful diet (eg, to feel better) were strongly associated with both fruit and vegetable intakes, and these associations were stronger for fruit. Extrinsic motivations were not associated with either fruit or vegetable intakes. APPLICATIONS: Dietary interventions based on a general 5-A-Day message may be more effective in increasing fruit intakes than vegetable intakes. Targeted interventions that focus specifically on vegetables are probably necessary. Intrinsic motives for eating a healthful diet should be key components of interventions to increase fruit and vegetable intakes. PMID- 9850111 TI - Variables affecting high school students' perceptions of school foodservice. AB - OBJECTIVE: To determine if student satisfaction with high school foodservice is directly related to participation in the foodservice. DESIGN: A valid and reliable survey was conducted in a variety of classes such as English, history, and health science in grades 9 through 12, representing students aged 13 through 19 years. Students were asked 38 questions concerning variety of food, food quality, foodservice staff, aesthetics of the serving and dining area, and demographics. SUBJECTS/SETTING: The study was conducted with 1,823 students from 9 schools representing 4 geographic regions. STATISTICAL ANALYSIS: Stepwise multiple regression was used to determine the independent variables (attributes desired by the students) that most highly correlated with the dependent variable (satisfaction with the school foodservice overall). RESULTS: Variables most highly correlated with overall satisfaction were variety of food offered, flavor of food, attractiveness of food on the serving line, staff smiling and greeting students, quality of food choices, choices that allow students to meet cultural and ethnic preferences, courteousness of the staff, and quality of ingredients. Variety of food offered was the best predictor of satisfaction. A statistically significant difference was found (P<.01) between groups that never ate school lunch and those that ate school lunch 3 to 5 times per week on dining ambiance, food quality, and staff. The results indicate that satisfaction with foodservice is associated with purchase behavior in school foodservice programs. APPLICATIONS: School foodservice and nutrition programs are critically important for providing nutrition to millions of our future leaders. Today it is not enough to prepare healthful, good-tasting food. High school students are sophisticated and are exposed at an early age to a variety of dining experiences including fast foods, ethnic cuisine, and fine dining. These factors have influenced the attributes students use to evaluate school foodservice. To maintain participation levels and financial stability, school foodservice professionals should evaluate student satisfaction with food quality, variety, and other variables that affect overall satisfaction and participation. These data may then be incorporated into continuous quality improvement and strategic planning. Marketing must be incorporated into the strategic plan to influence student participation. PMID- 9850110 TI - The 'age+5' rule: comparisons of dietary fiber intake among 4- to 10-year-old children. AB - OBJECTIVE: To determine children's sources of dietary fiber and to identify the food group choices made by those who met the "age+5" rule, which recommends that children daily consume an amount of fiber that is equal to their age plus an additional 5 g fiber. DESIGN: This study used 24-hour dietary recalls and 1-day food records to assess the nutrient intake and food group choices of children who did and did not meet the recommendations of the age+5 rule. SUBJECTS: The 1989 1991 US Department of Agriculture Continuing Survey of Food Intakes by Individuals provided the study sample of 603 children between the ages of 4 and 6 years and 782 children between the ages of 7 and 10 years. STATISTICAL ANALYSES: Differences in nutrient and food group intakes between age categories were determined by t tests. RESULTS: Only 45% of 4- to 6-year-olds and 32% of 7- to 10 year-olds consumed adequate fiber to meet the age+5 rule. Those who met the age+5 rule did so by consuming significantly more high- and low-fiber breads and cereals, fruits, vegetables, legumes, nuts, and seeds. Children with low fiber intakes had significantly higher energy-adjusted intakes of fat and cholesterol, whereas those who met the age+5 rule had significantly higher energy-adjusted intakes of dietary fiber, vitamins A and E, folate, magnesium, and iron. APPLICATIONS: The majority of the children had low intakes of dietary fiber, suggesting that they are at risk for future chronic disease. Parents and school foodservice personnel should strive to offer fiber-rich foods to children so their acceptance and consumption of them will be increased. PMID- 9850112 TI - Making hot lunch cool: marketing strategies for school foodservices. PMID- 9850113 TI - Impact of breakfast consumption on nutritional adequacy of the diets of young adults in Bogalusa, Louisiana: ethnic and gender contrasts. AB - OBJECTIVE: To evaluate the impact of breakfast consumption patterns on the nutritional adequacy of diets of young adults and determine possible ethnic and gender differences. DESIGN AND SETTING: Cross-sectional survey of young adults in Bogalusa, La. SUBJECTS: Twenty-four-hour dietary recalls were collected from October 1988 through October 1991 on 504 young adults (mean age=23 years, 58% women, 70% white). STATISTICS: Analysis of variance and logistic regression techniques were used to investigate the relationship of breakfast consumption, ethnicity, and gender on dietary adequacy. The P values are from an analysis of variance model that adjusted for gender and ethnicity. RESULTS: Thirty-seven percent of young adults skipped breakfast. Of those who ate breakfast, 75% ate at home, 10% ate a fast-food breakfast, and 15% reported other sources. Mean energy intake from breakfast was 485 kcal; men consumed more energy than women (P<.001), and blacks consumed more energy than whites (P<.01). The breakfast meal provided an average of 13% of energy from protein, 55% from carbohydrate, 14% from sucrose, 34% from fat, and 12% from saturated fat. Whites consumed a breakfast higher in carbohydrate and sucrose than blacks, who consumed a breakfast higher in fat and saturated fat. Variations in breakfast foods consumed explained the racial differences in the nutrient composition of the breakfast meal. Young adults who skipped breakfast had lower total daily intakes of energy (P<.0001), protein per 1,000 kcal (P<.05), and saturated fat per 1,000 kcal (P<.01) than those who consumed breakfast. For all vitamins and minerals studied, a higher percentage of young adults who skipped breakfast did not meet two thirds of the Recommended Dietary Allowance than those who consumed a breakfast. APPLICATIONS: Encouraging consumption of breakfast, along with selection of more healthful breakfast food choices or snacks that are culturally appropriate, may be important strategies for improving the nutritional quality of young adults' diets. PMID- 9850114 TI - Assessing adherence to a rotary diversified diet, a treatment for 'environmental illness'. AB - OBJECTIVES: To develop and test a method to assess adherence to a rotary diversified diet (RDD), a treatment for environmental illness, which is a putative disorder characterized by multiple sensitivities to foods, chemicals, or inhalants. The RDD requires the elimination of prohibited foods and rotation of remaining nonprohibited foods and their "food families" within a 4- to 7-day cycle. The regimen has yet to be validated to the satisfaction of the scientific community. DESIGN: Details of the 2 components of the RDD prescription, elimination and rotation, were documented, and a food record method of assessing adherence was developed. Adherence to the RDD was then assessed in a cohort of women who were enrolled in a larger prospective study. Test-retest reliability of the adherence assessment method was determined by calculating ratings twice on the same set of patient food records, with 1 week between trials. SUBJECTS/SETTING: All patients were contacted through a private environmental medicine clinic in Toronto, Canada. Eight patients provided the food records needed for development of the method; adherence was then assessed in 22 women aged 25 to 67 years. STATISTICAL ANALYSES: Means, standard deviations, and 95% confidence intervals for adherence ratings were calculated. The reliability of the adherence assessment method was determined by calculating Pearson correlation coefficients for adherence ratings from each trial. A paired t test was also used to determine if the mean differences in ratings between trials were significant. RESULTS: Patients experienced difficulties following both components of the RDD: 37% to 44% of foods consumed were either prohibited or allowed, but were consumed on the incorrect day. The adherence assessment method was found to have high levels of reliability. APPLICATIONS: The adherence assessment method can be used in future evaluations of the RDD, although further testing of the method is recommended. Increased involvement of dietitians with patients diagnosed with environmental illness is recommended. PMID- 9850115 TI - Productive aging: a quality of life agenda. AB - Although the fact that, in general, the US population is living longer is said to be the public health success story of the 20th century, the boom in the population aged 65 years and older is having and will continue to have major implications for all age groups as well as for the nation's health, social, and economic institutions. The concept of productive aging can provide guidance in addressing these implications. The assumptions of productive aging reflect today's reality that older people are repositories of wisdom and experience and important assets for society; relatively healthy, with the potential for maintaining that condition until late in life; capable of making economic and social contributions that benefit themselves, their families, and their communities; and in need of purposeful and meaningful roles and activities in life. A productive aging activity model emphasizes involvement in paid work, volunteerism, education, fitness and exercise, leisure and travel, advocacy and political action, and consumerism. This article discusses the concept of productive aging, a related activity model, and qualitative focus group research undertaken in conjunction with the 1995 White House Conference on Aging. PMID- 9850116 TI - Lessons learned about adolescent nutrition from the Minnesota Adolescent Health Survey. AB - In 1986-1987, more than 30,000 adolescents completed the Minnesota Adolescent Health Survey, a comprehensive assessment of adolescent health status, health behaviors, and psychosocial factors. Although the survey included relatively few items on nutrition-related issues, a wealth of knowledge about adolescent nutrition was gained. Lessons learned from a decade of subsequent analyses of data collected in the survey and implications for working with youth are summarized in this article. Major concerns identified included high prevalence rates of inadequate intake of fruits, vegetables, and dairy products; unhealthful weight-control practices; and overweight status. For example, inadequate fruit intake was reported by 28% of the adolescents and inadequate vegetable intake was reported by 36%. Among female adolescents, 12% reported chronic dieting, 30% reported binge eating, 12% reported self-induced vomiting, and 2% reported using diuretics or laxatives. Some of the risk factors for inadequate food intake patterns or unhealthful weight-control practices included low socioeconomic status, minority status, chronic illness, poor school achievement, low family connectedness, weight dissatisfaction, overweight, homosexual orientation among male adolescents, and use of health-compromising behaviors. To improve adolescent eating behaviors, the results suggest a need for innovative outreach strategies that include educational and environmental approaches. Dietitians play a key role in developing interventions and promoting research in the field of adolescent nutrition. PMID- 9850117 TI - Nutrition supplementation enables elderly residents of long-term-care facilities to meet or exceed RDAs without displacing energy or nutrient intakes from meals. PMID- 9850118 TI - Effect of participation in congregate-site meal programs on the energy and nutrient intakes of Hispanic seniors. PMID- 9850119 TI - Comparison of standard and portion-assisted responses to a brief fruit and vegetable food frequency questionnaire in a Hispanic WIC population. PMID- 9850120 TI - Students cluster into 4 groups according to the factors influencing their dietary intake. PMID- 9850121 TI - Sensory characteristics and acceptability of lactose-reduced baked custards made with an egg substitute. PMID- 9850122 TI - President's page: Nutrition messages on food packages--location, location, location. PMID- 9850123 TI - On genes, individuals, society, and epidemiology. PMID- 9850124 TI - Racial differences in lens opacities: the Salisbury Eye Evaluation (SEE) project. AB - The purpose of this study was to determine racial differences in the prevalence of different types of lens opacities and cataract surgery. Between 1993 and 1995, the Salisbury Eye Evaluation (SEE) Project enrolled a representative sample of 2,520 community-dwelling persons aged 65-84 years in Salisbury, Maryland, 26.4% of whom were African-American. Participants received a full eye examination, and photographs were taken for documentation of lens status. Photographs were graded using a standardized grading system for the presence of cortical, nuclear, or posterior subcapsular cataract (PSC) opacification in at least one eye. The odds of having cortical opacities were 4.0 times greater among African Americans than among Caucasians (95% confidence interval (CI) 3.3-4.8). Caucasians were significantly more likely to have nuclear opacities (odds ratio = 2.1, 95% CI 1.7 2.6) and PSC opacities (odds ratio = 2.5, 95% CI 1.7-3.6). The odds of cataract surgery were 2.8 times higher among Caucasians, but these differences did not explain the differences in the prevalence of different types of lens opacities by racial group. With lower rates of nuclear and PSC opacities than Caucasians, African Americans may have a lower demand for cataract surgery. However, even with these differences, there is still significant unnecessary loss of vision due to cataract among older African Americans, for whom programs to ensure access to surgical care are indicated. PMID- 9850125 TI - Lifetime exposure to environmental tobacco smoke among urban women: differences by socioeconomic class. AB - This study sought to determine cumulative lifetime exposure to environmental tobacco smoke (ETS) among urban women in relation to sociodemographic factors. In a population survey carried out in Geneva, Switzerland, during 1993-1995, a representative sample of 1,883 women aged 35-74 years answered interview questions on lifetime ETS exposure. Exposed women were defined as those who had spent at least 1 hour daily in a smoky environment during 1 or more years. The prevalence of current ETS exposure was 31.0% among 1,458 never or former smokers. Lifetime prevalence was 58.3% among 1,061 never smokers. The home (42.1%) and the workplace (39.6% of employed women) were the most frequent sources of ETS exposure, leisure time activity being a secondary source. Throughout a lifetime, work accounted for the greatest average intensity of exposure (on average, 19 hours of exposure per week), while the longest duration of exposure (on average, 18 years) was in the home. Cumulative lifetime exposure (intensity (in hours/week) x duration) from all sources combined was 308 hours/week-years, which can correspond to 30.8 hours/week over a period of 10 years or 20.5 hours/week over a period of 15 years. Women from low socioeconomic classes had more intense and longer exposures than women from higher socioeconomic classes, mainly because of work exposure. Both the intensity and the duration of lifetime ETS exposure were greater than previously suspected. Reduction of ETS exposure in the workplace should be a public health priority. PMID- 9850126 TI - Completeness of ascertainment of prenatal smoking using birth certificates and confidential questionnaires: variations by maternal attributes and infant birth weight. PRAMS Working Group. Pregnancy Risk Assessment Monitoring System. AB - Birth certificate data frequently are used to monitor the prevalence of smoking during pregnancy. The authors used a two-sample capture-recapture method to estimate the completeness of ascertainment of prenatal smoking on birth certificates and on confidential questionnaires in six US states. Completeness of ascertainment was also examined according to maternal attributes and infant birth weight. The samples included white women who delivered a live infant between 1993 and 1995 in one of six states (Alabama, Alaska, Georgia, Maine, South Carolina, or West Virginia) and who responded to a questionnaire mailed to them 2-6 months postpartum as part of the Pregnancy Risk Assessment Monitoring System. State specific sample sizes ranged from 2,647 to 4,795. The completeness of ascertainment ranged from 70.6% to 82.0% using birth certificates and from 86.2% to 90.3% using confidential questionnaires. In all six states, the birth certificates' completeness of ascertainment varied by maternal education and infant birth weight, and the questionnaires' completeness varied by maternal age. Both birth certificates and questionnaires underestimated the true extent of smoking during pregnancy among these white women. Differential reporting by birth weights recorded on birth certificates would result in an overestimated association between low birth weight and prenatal smoking. PMID- 9850127 TI - Timing of menopause, reproductive years, and bone mineral density: a cross sectional study of postmenopausal Japanese women. AB - Age at menopause has been found to be associated positively with bone mineral density, and age at menarche has been found to be associated negatively with bone mineral density. However, there have been few studies on the relations of timing of menopause and length of the reproductive period with bone mineral density. The purpose of this study was to examine the relations of timing of menopause and reproductive years (calculated as age at menopause minus age at menarche) with mineral density of the second metacarpal bone in postmenopausal Japanese women. The study population consisted of 1,035 naturally menopausal women aged 40-70 years who were screened in 1996-1997. Using computed x-ray densitometry, the authors measured bone mineral density by analyzing radiographic films of the right second metacarpal bone. Using the women with early menopause (age < 49 years) as the reference group and adjusting for age, subjects with late menopause were at decreased risk for low bone mineral density (odds ratio (OR) = 0.69, 95% confidence interval (CI) 0.49-0.97). After adjustment for additional covariates (grip strength, physical activity, body mass index, smoking, and calcium intake), the association was unchanged (OR = 0.70, 95% CI 0.50-0.99). Postmenopausal women with more reproductive years (> or = 40 years) were at decreased risk for low bone mineral density compared with those with fewer reproductive years, after adjustment for age (OR = 0.73, 95% CI 0.40-1.30) and potentially confounding factors (OR = 0.76, 95% CI 0.41-1.37); the p-value for trend was not statistically significant. In multiple linear regression analysis, early menopause and fewer reproductive years were independent predictors of low bone mineral density. In this study, postmenopausal Japanese women who had a late menopause and more reproductive years were at decreased risk for low bone mineral density, and may therefore be less prone to osteoporosis. PMID- 9850128 TI - Relation of self-image to body size and weight loss attempts in black women: the CARDIA study. Coronary Artery Risk Development in Young Adults. AB - It has been suggested that the prevalence of obesity in black women is high partly because self-image in black women is not strongly dependent on body size. To determine associations between self-image, body size, and dieting behavior among black women, the authors assessed an Appearance Evaluation Subscale (AES) score (range, 1-5), a Body Image Satisfaction (BIS) score (range, 2-11), and reported dieting behavior in a population-based sample of 1,143 black women aged 24-42 years from the fourth follow-up examination (1992-1993) of the Coronary Artery Risk Development in Young Adults (CARDIA) Study. Lower AES and BIS scores indicate poorer self-image and lower body size satisfaction, respectively. After adjustment for age, education, smoking, and physical activity, women in the lowest, middle, and highest tertiles of body mass index (weight (kg)/height (m)2) had mean AES scores of 3.7, 3.3, and 2.9, respectively (p < 0.001), and mean BIS scores of 7.8, 6.7, and 5.9, respectively (p < 0.001). After additional control for body mass index as a continuous variable, both AES and BIS scores were inversely related to ever dieting, current dieting, and previous weight loss of 10 pounds (4.5 kg) or more in all tertiles of body mass index. These results suggest that among black women, a higher body mass index is associated with poorer self-image and lower body size satisfaction and that these perceptions may be an avenue to promoting weight control. PMID- 9850129 TI - Adult height, stroke, and coronary heart disease. AB - An inverse relation between adult height and risk of coronary heart disease (CHD) has been reported in many studies, but the association between adult height and stroke remains uncertain. The authors examined the relation between adult height and risk of stroke and CHD in a prospective study of 7,735 men drawn from general medical practices in 24 towns in England, Wales, and Scotland. The men were followed up for an average of 16.8 years (range, 15.5-18.0 years) between 1978 and 1995. During this period, there were 351 major stroke events (63 fatal, 288 nonfatal) and 1,093 major CHD events (465 fatal, 628 nonfatal). The mean height of the men was 173.3 cm. Total stroke risk was increased only in the men who fell into the lowest quintile of the height distribution (<167.7 cm), with little difference being seen between the other groups. When data were examined separately for fatal and nonfatal events, no relation was seen with nonfatal stroke. An apparent inverse association was seen with fatal stroke, even after adjustment for a wide range of confounding variables, but the number of deaths was small and the trend was not statistically significant (p = 0.17). By contrast, a significant inverse relation was seen between height and risk of major CHD events: Risk decreased progressively with increasing height, even after full adjustment (highest quintile vs. lowest: relative risk (RR) = 0.74, 95% confidence interval (CI) 0.59-0.91; test for trend: p < 0.001). A stronger inverse association was seen with nonfatal CHD events (RR = 0.64, 95% CI 0.49 0.84) than with fatal CHD events (RR = 0.82, 95% CI 0.60-1.11). This study confirms the finding of an inverse association between height and CHD. The inverse association seen for fatal stroke but not nonfatal stroke suggests that height may be related to specific subtypes of stroke. There are different patterns of association between height and stroke and height and CHD. If the apparent association between short stature and increased risk of fatal stroke is confirmed in other prospective studies, this would suggest that different mechanisms underlie the effects of height on stroke and CHD. PMID- 9850130 TI - Long term mortality and morbidity related to degree of damage following the 1998 earthquake in Armenia. AB - To assess the relation of increased mortality and morbidity to personal loss and damage following the 1988 earthquake in Armenia, the authors conducted a prospective study of mortality and a nested case-control analysis of incident morbidity. Employees of the Armenian Ministry of Health and their immediate families (n = 35,043) who survived the disaster formed the study population. Two sets of interviews with the employees, carried out over a period of 4 years of follow-up, were used as the primary source of data for this study. The highest numbers of deaths from all causes and from heart disease were observed within the first 6 months following the earthquake. The nested case-control analysis of 483 cases of newly reported heart disease and 482 matched non-heart-disease controls revealed that people with increasing levels of loss of material possessions and family members had significant increases in heart disease risk (odds ratios for "loss scores" of 1, 2, and 3 were 1.3, 1.8, and 2.6, respectively). The findings were similar with regard to the relation of damage and loss to newly reported hypertension, diabetes mellitus, and arthritis. The findings of this study support the hypothesis that longer term increased rates of heart disease and chronic disease morbidity following an earthquake are related to the intensity of exposure to disaster-related damage and losses. People sustaining such losses should be closely monitored for increased long term morbidity. PMID- 9850131 TI - Assessment of trans-fatty acid intake with a food frequency questionnaire and validation with adipose tissue levels of trans-fatty acids. AB - Past studies of the association of trans-fatty acid intake with coronary heart disease have been hindered by the lack of a database on the trans-fatty acid content of various foods. The authors used new data from the US Department of Agriculture to estimate trans-fatty acid intake using a self-administered food frequency questionnaire (FFQ), and they assessed the validity of the FFQ by comparing the dietary estimates with trans-fatty acid concentrations in adipose tissue. The 1996 study included 27 women and 24 men aged 51-78 years. The mean consumption of total trans-fatty acids estimated from the FFQ was 2.24 g per day and 5% of total dietary fat. The mean concentration of total trans-fatty acids in buttock adipose tissue was 4.7% of total fatty acids. Pearson correlations between total dietary intake of trans-fatty acids and total trans-fatty acid levels in adipose tissue were 0.67 (95% confidence interval (CI) 0.36-0.84) among men and 0.58 (95% CI 0.26-0.79) among women. After adjustment for energy intake, age, and body mass index, the correlation coefficients were 0.76 (95% CI 0.51 0.89) among men and 0.52 (95% CI 0.17-0.75) among women. The FFQ validated in this study is an important new tool for assessing usual intake of trans-fatty acids. PMID- 9850132 TI - Impact of influenza on acute cardiopulmonary hospitalizations in pregnant women. AB - This study sought to quantify influenza-related serious morbidity in pregnant women, as measured by hospitalizations for or death from selected acute cardiopulmonary conditions during predefined influenza seasons. The study population included women aged 15-44 years who were enrolled in the Tennessee Medicaid program for at least 180 days between 1974 and 1993. In a nested case control study, 4,369 women with a first study event during influenza season were compared with 21,845 population controls. The odds ratios associated with study events increased from 1.44 (95% confidence interval (CI) 0.97-2.15) for women at 14-20 weeks' gestation to 4.67 (95% CI 3.42-6.39) for those at 37-42 weeks in comparison with postpartum women. A retrospective cohort analysis, which controlled for risk factors identified in the case-control study, identified 22,824 study events during 1,393,166 women-years of follow-up. Women in their third trimester without other identified risk factors for influenza morbidity had an event rate of 21.7 per 10,000 women-months during influenza season. Approximately half of this morbidity, 10.5 (95% CI 6.7-14.3) events per 10,000 women-months, was attributable to influenza. Influenza-attributable risks in comparable nonpregnant and postpartum women were 1.91 (95% CI 1.51-2.31) and 1.16 (95% CI -0.09 to 2.42) per 10,000 women-months, respectively. The data suggest that, out of every 10,000 women in their third trimester without other identified risk factors who experience an average influenza season of 2.5 months, 25 will be hospitalized with influenza-related morbidity. PMID- 9850133 TI - Explosive school-based measles outbreak: intense exposure may have resulted in high risk, even among revaccinees. AB - Even high levels of measles vaccination coverage have not always prevented outbreaks of measles spread by airborne transmission. It has been suggested that a large inoculum might increase vaccine failure risk. Airbome transmission might occasionally entail a large measles inoculum. The epidemiologic relevance of measles among properly vaccinated persons (i.e., those vaccinated after 15 months of age and with live attenuated virus) is increased when they become contagious. The authors studied inoculum intensities as measured by proxy variables and the contagiousness of properly vaccinated persons who contracted measles among 51 measles patients infected in one school, at home, or elsewhere, utilizing preexisting records of measles cases and 214 healthy controls from an explosive school outbreak that occurred in a rural Finnish municipality in 1989. One "super spreader" infected 22 others in one day, including eight once-vaccinated students and one twice-vaccinated student, probably during an assembly of 144 students in a poorly ventilated hallway with no sunlight. Those infected later at home had high measles risk, even if they were revaccinees. When siblings shared a bedroom with a measles case, a 78 percent risk (seven out of nine children) was observed among vaccinees. Vaccinees had approximately 2 days' shorter incubation time than unvaccinated persons. Vaccinated and unvaccinated students were equally able to infect their siblings. Total protection against measles might not be achievable, even among revaccinees, when children are confronted with intense exposure to measles virus. PMID- 9850134 TI - Determinants of dengue 2 infection among residents of Charters Towers, Queensland, Australia. AB - Dengue fever is caused by one of the four serotypes of the dengue virus and is transmitted by the urban mosquito Aedes aegypti. In 1993, the city of Charters Towers in the tropical north of Australia experienced an epidemic caused by the dengue 2 virus. A cross-sectional sample of 1,000 people was assessed for determinants of recent symptomatic dengue infection. After exclusion of people with prior exposure to dengue 2, a study group of 797 persons, including 196 patients with recent infection, were evaluated. Stepwise logistic regression analysis identified four determinants of infection: the presence of a case of dengue fever within two residential blocks (odds ratio (OR) = 3.61, 95% confidence interval (CI) 2.56-5.10), house screening (OR = 0.60, 95% CI 0.40 0.89), the presence of a water tank within two residential blocks (OR = 1.51, 95% CI 1.02-2.22), and the use of knockdown insecticide (OR = 1.75, 95% CI 1.22 2.51). Classification and Regression Tree analysis identified a group of 152 individuals in whom the prevalence of dengue infection was 50%. These people lived within two blocks of a suspected dengue fever case, did not have house screening, and used knockdown sprays. If dengue had not occurred within two residential blocks, there were no additional factors that significantly influenced the prevalence of dengue fever. Control of dengue epidemics should involve attempts to geographically contain the spread of infection, use of house screening, and the removal of mosquito breeding sites such as water tanks. PMID- 9850135 TI - Heterogeneity of the baseline risk within patient populations of clinical trials: a proposed evaluation algorithm. AB - In this paper, the authors present an evaluation algorithm for systematic assessment of the observed heterogeneity in disease risk within trial populations. Predictive models are used to estimate the predicted patient hazards, the odds of having an event in the upper risk quartile (ODU) and the lower risk quartile (ODL), and the odds ratio (rate ratio for time-to-event analyses) for having an event in the upper risk quartile versus the lower risk quartile (extreme quartile odds ratio (EQuOR) and extreme quartile rate ratio (EQuRR)). The ranges for these metrics depend on the extent to which predictors of the outcome of interest exist and are known and the extent to which data are collected in the trial, as well as on the eligibility criteria and the specific patients who are actually enrolled. ODU, ODL, and EQuOR values are used to systematically interpret the results for patients at different levels of risk, to evaluate generalizability, and to determine the need for subgroup analyses. Individual data for five outcomes from three trials (n = 842, 913, and 1,001, respectively) are used as examples. Observed EQuOR values ranged from 1.5 (very little predicted heterogeneity) to 59 (large heterogeneity). EQuRR values ranged from 2 to 46. ODU values ranged from 0.24 to 3.19 (generally high risk), and ODL values ranged from 0.01 (clinically negligible risk) to 0.16 (clinically meaningful risk). The algorithm may also be used for comparing diverse trials (e.g., in meta-analyses) and used prospectively for designing future trials, as shown in simulations. PMID- 9850136 TI - Slopes of a receiver operating characteristic curve and likelihood ratios for a diagnostic test. AB - This paper clarifies two important concepts in clinical epidemiology: the slope of a receiver operating characteristic (ROC) curve and the likelihood ratio. It points out that there are three types of slopes in an ROC curve--the tangent at a point on the curve, the slope between the origin and a point on the curve, and the slope between two points on the curve. It also points out that there are three types of likelihood ratios that can be defined for a diagnostic test that produces results on a continuous scale--the likelihood ratio for a particular single test value, the likelihood ratio for a positive test result, and the likelihood ratio for a test result in a particular level or category. It further illustrates mathematically and empirically the following three relations between these various definitions of slopes and likelihood ratios: 1) the tangent at a point on the ROC curve corresponds to the likelihood ratio for a single test value represented by that point; 2) the slope between the origin and a point on the curve corresponds to the positive likelihood ratio using the point as a criterion for positivity; and 3) the slope between two points on the curve corresponds to the likelihood ratio for a test result in a defined level bounded by the two points. The likelihood ratio for a single test value is considered an important parameter for evaluating diagnostic tests, but it is not easily estimable directly from laboratory data because of limited sample size. However, by using ROC analysis, the likelihood ratio for a single test value can be easily measured from the tangent. It is suggested that existing ROC analysis software be revised to provide estimates for tangents at various points on the ROC curve. PMID- 9850137 TI - Re: "Tobacco as a cause of lung cancer: some reflections". PMID- 9850138 TI - Re: "Lactation history and breast cancer risk". PMID- 9850139 TI - Ischemic and excitotoxic brain injury is enhanced in mice lacking the p55 tumor necrosis factor receptor. AB - Ischemic and excitotoxic insults to the brain induce rapid production of tumor necrosis factor-alpha (TNF), but the role of TNF in neuronal responses to brain injury are unclear. Two different TNF receptors (p55 and p75) are expressed in neurons and glia. To understand the role of TNF in brain injury, we generated mice that lack p55, p75, or both receptors. We report that neuronal damage after focal cerebral ischemia-reperfusion is significantly increased in mice lacking p55 receptors (85+/-7 mm3 infarct volume; mean +/- SD) compared with wild-type mice (70+/-8 mm3) and mice lacking p75 receptors (72+/-6 mm3). Moreover, mice lacking p55 receptors exhibited increased degeneration of CA3 hippocampal neurons after administration of the excitotoxin kainic acid compared with wild-type mice and mice lacking p75 receptors. When taken together with recent data showing that TNF can prevent apoptosis of cultured neurons exposed to oxidative and metabolic insults, our findings suggest that TNF plays a neuroprotective role after acute brain insults. PMID- 9850140 TI - Distribution volume of radiolabeled large neutral amino acids in brain tissue. AB - Variations in the cerebellum to plasma ratio at late times in 6-[18F]fluoro-L DOPA studies are shown to be consistent with competitive binding of large neutral amino acids for a common transporter in the blood-brain barrier and the stability of brain tissue large neutral amino acid level in the presence of plasma level changes. The distribution volume of an inert large neutral amino acid can be estimated from plasma and tissue large neutral amino acid levels and apparent half-saturation concentrations (Km) of the transporter in the blood-brain barrier. Stability of brain large neutral amino acid levels is supported by literature findings and can be explained by high saturation of the large neutral amino acid transporter at physiologic conditions. PMID- 9850141 TI - Moderate hypothermia reduces hypotensive, but not hypercapnic vasodilation of pial arterioles in rats. AB - Two types of acid-base strategies are available for the blood gas management of patients during hypothermia: alpha-stat and pH-stat management. However, the more suitable strategy for therapeutic hypothermia is unclear. We studied the effects of hypothermia (30 degrees C) and acid-base management on reactivity to hypercapnia and hypotension in rat pial arterioles, using a closed cranial window. The baseline diameter during hypothermia decreased in the alpha-stat (PaCO2 was maintained at 35 mm Hg when measured at 37 degrees C, n = 8), but not in the pH-stat (PaCO2 was maintained at 35 mm Hg when corrected to the animal's actual temperature, n = 7). Vasodilation induced by hypotension was significantly reduced in hypothermic groups compared with the normothermic group (n = 7), whereas responses to hypercapnia were preserved. Moreover, hypotensive vasodilation was more attenuated in the pH-stat, than the alpha-stat, management. These findings show that moderate hypothermia and acid-base management alter cerebrovascular autoregulation. PMID- 9850142 TI - Tissue at risk of infarction rescued by early reperfusion: a positron emission tomography study in systemic recombinant tissue plasminogen activator thrombolysis of acute stroke. AB - Thrombolytic therapy of acute ischemic stroke can be successful only as long as there is penumbral tissue perfused at rates between the thresholds of normal function and irreversible structural damage, respectively. To determine the proportion of tissue at risk of infarction, cerebral perfusion was studied in 12 patients with acute ischemic stroke who underwent treatment with systemic recombinant tissue plasminogen activator (0.9 mg/kg body weight according to National Institute of Neurological Disorders and Stroke protocol) within 3 hours of onset of symptoms, using [15O]-H2O positron emission tomography (PET) before or during, and repeatedly after thrombolysis. The size of the regions of critically hypoperfused gray matter were identified on the initial PET scans, and changes of perfusion in those areas were related to the clinical course (followed by the National Institutes of Health stroke scale) and to the volume of infarcted gray matter demarcated on magnetic resonance imaging 3 weeks after the stroke. Whereas the initial clinical score was unrelated to the size of the ischemic area, after 3 weeks there was a strong correlation between clinical deficit and volume size of infarcted gray matter (Spearman's rho, 0.96; P < 0.001). All patients with a severely hypoperfused (< 12 mL/100 g/min) gray matter region measuring less than 15 mL on first PET showed full morphologic and clinical recovery (n = 5), whereas those with ischemic areas larger than 20 mL developed infarction and experienced persistent neurologic deficits of varying degree. Infarct sizes, however, were smaller than expected from previous correlative PET and morphologic studies of patients with acute stroke: only 22.7% of the gray matter initially perfused at rates below the conventional threshold of critical ischemia became necrotic. Actually, the percentage of initially ischemic voxels that became reperfused at almost normal levels clearly predicted the degree of clinical improvement achieved within 3 weeks. These sequential blood flow PET studies demonstrate that critically hypoperfused tissue can be preserved by early reperfusion, perhaps related to thrombolytic therapy. The results correspond with experimental findings demonstrating the prevention of large infarcts by early reperfusion to misery perfused but viable tissue. PMID- 9850143 TI - Effect of cortical spreading depression on the levels of mRNA coding for putative neuroprotective proteins in rat brain. AB - Previous studies have demonstrated that cortical spreading depression (CSD) induces neuronal tolerance to a subsequent episode of ischemia. The objective of the present investigation was to determine whether CSD alters levels of mRNA coding for putative neuroprotective proteins. Unilateral CSD was evoked in male Wistar rats by applying 2 mol/L KCl over the frontal cortex for 2 hours. After recovery for 0, 2, or 24 hours, levels of several mRNA coding for neuroprotective proteins were measured bilaterally in parietal cortex using Northern blot analysis. Levels of c-fos mRNA and brain-derived neurotrophic factor (BDNF) mRNA were markedly elevated at 0 and 2 hours, but not 24 hours after CSD. Tissue plasminogen activator (tPA) mRNA levels were also significantly increased at 0 and 2 hours, but not 24 hours after CSD. Levels of the 72-kDa heat-shock protein (hsp72) mRNA were not significantly increased by CSD, except for a small elevation (20%) at 2 hours recovery. Levels of the 73-kDa heat-shock cognate (hsc73) mRNA were slightly, but significantly, increased at 2 and 24 hours of recovery. Finally, levels of mRNA for protease nexin-1 and glutamine synthetase were not significantly altered by CSD at any time studied. The current results support the hypothesis that neuronal tolerance to ischemia after CSD may be mediated by increased expression of FOS, BDNF, or tPA, but not by increased expression of hsp72, hsc73, nexin-1, or glutamine synthetase. PMID- 9850144 TI - Astrocyte regulation of endothelial tissue plasminogen activator in a blood-brain barrier model. AB - Expression of tissue plasminogen activator (tPA) substantially determines endothelial-dependent fibrinolysis. We used a blood-brain barrier (BBB) model to analyze regulation of brain capillary endothelial tPA and its inhibitor, plasminogen activator inhibitor-1 (PAI-1). This model consists of coculture of murine astrocytes with bovine brain capillary endothelial cells grown as capillary-like structures (CS); after 1 week, astrocytes become extensively associated with CS, and the BBB-associated enzyme gamma-glutamyl transpeptidase is present. We measured tPA and PAI-1 mRNA and tPA activity in this model. Reverse transcription-polymerase chain reaction (RT-PCR) studies showed similar tPA and PAI-1 mRNA levels after 1 day mono-culture (endothelial cells only) versus astrocyte-endothelial coculture preparations. After 7 days (i.e., when elements of the BBB are present), astrocyte-endothelial cocultures (compared with endothelial mono-cultures) showed a 50.7%+/-27.1% (mean +/- SD) reduction in tPA mRNA (P < 0.03) and a 183.3%+/-86.9% increase in PAI-1 mRNA expression (P < 0.02). Moreover, 7-day cocultures demonstrated reduced tPA activity compared with mono-cultures (14.6+/-2.9 IU/mL versus 30.2+/-7.7 IU/mL, P < 0.01); 1-day cocultures and mono-cultures had similar tPA activity. These findings demonstrate that astrocytes regulate brain capillary endothelial expression of tPA when elements of the BBB phenotype are present in this model. These data suggest an important role for astrocytes in the regulation of brain capillary endothelial fibrinolysis. PMID- 9850145 TI - Attenuated c-fos mRNA induction after middle cerebral artery occlusion in CREB knockout mice does not modulate focal ischemic injury. AB - To elucidate the mechanism of ischemia-induced signal transduction in vivo, we investigated the effect of the targeted disruption of the alpha and delta isoforms of the cAMP-responsive element-binding protein (CREB) on c-fos and heatshock protein (hsp) 72 gene induction. Permanent focal ischemia was induced by occlusion of the middle cerebral artery of the CREB mutant mice (CREB(-/-), n = 5) and the wild-type mice (n = 6). Three hours after onset of ischemia, the neurologic score was assessed and pictorial measurements of ATP and cerebral protein synthesis (CPS) were carried out to differentiate between the ischemic core (where ATP is depleted), the ischemic penumbra (where ATP is preserved but CPS is inhibited), and the intact tissue (where both ATP and CPS are preserved). There were no significant differences in neurologic score or in ATP, pH, and CPS between the two groups, suggesting that the sensitivity of both strains to ischemia is the same. Targeted disruption of the CREB gene significantly attenuated c-fos gene induction in the periischemic ipsilateral hemisphere but had no effect on either c-fos or hsp72 mRNA expression in the penumbra. The observations demonstrate that CREB expression, despite its differential effect on c-fos, does not modulate acute focal ischemic injury. PMID- 9850146 TI - Deficiency of intercellular adhesion molecule 1 attenuates microcirculatory disturbance and infarction size in focal cerebral ischemia. AB - Recent evidence has shown crucial roles for cell-adhesion molecules in inflammation-induced rolling, adhesion, and accumulation of neutrophils in tissue. Intercellular adhesion molecule-1 (ICAM-1) is one of these adhesion molecules. Previous studies have shown marked reduction in the size of infarction after focal cerebral ischemia by depletion of granulocytes and administration of the antibody against ICAM-1. In the present study we investigated the role of ICAM-1 in the size of ischemic lesions, accumulation of granulocytes, and microcirculatory compromise in focal cerebral ischemia by using ICAM-1-knockout mice. Ischemic lesions were significantly mitigated in knockout mice after permanent and transient focal ischemia, even though the number of granulocytes in the infarcted tissue was almost the same between knockout and wild-type mice. Depletion of granulocytes further decreased the size of ischemic lesions after transient focal ischemia in ICAM-1-knockout mice. Microcirculation was reduced after focal ischemia, but it was better preserved in the cerebral cortex of knockout mice than that of wild-type mice. The present study demonstrated that ICAM-1 played a role in microcirculatory failure and subsequent development and expansion of infarction after focal cerebral ischemia. However, it is highly unlikely that ICAM-1 played a key role in accumulation of granulocytes after focal cerebral ischemia. PMID- 9850147 TI - Early postischemic dantrolene-induced amelioration of poly(ADP-ribose) polymerase related bioenergetic failure in neonatal rat brain slices. AB - In the infant brain, ischemia-induced ionic and enzyme mechanisms may independently lead to cell death by energy depletion: resequestration of calcium mobilized from intracellular stores consumes ATP, and activated poly(ADP-ribose) polymerase (PARP) uses oxidized nicotinamide adenine dinucleotide to form polyADP ribosyl nuclear proteins associated with DNA damage. Using 31P nuclear magnetic resonance spectroscopy, we have monitored intracellular pH and cellular energy metabolites in ex vivo neonatal rat cerebral cortex before, during, and after substrate and oxygen deprivation. In an insult that exhibited secondary energy failure and apoptosis we identified a relative 25% augmentation of high-energy phosphates at the end of recovery when the ryanodine-receptor antagonist, dantrolene, was introduced in the early (0- to 40-minute) but not late (40- to 120-minute) stage of recovery (P < 0.05). In contrast to the absence of a late dantrolene-sensitive effect, inhibition of PARP with 3-methoxybenzamide was as effective (P < 0.05) as early dantrolene, even when introduced after a 40-minute delay. The dantrolene and 3-methoxybenzamide effects on high-energy phosphates were not additive, rather the early dantrolene-sensitive effect nullified the potential 3-methoxybenzamide effect. Therefore, in this vascular-independent neonatal preparation, postischemic mobilization of calcium from intracellular stores is associated with PARP-related energy depletion. Inhibition of either of these processes confers improved postischemic bioenergetic recovery in the developing brain. PMID- 9850148 TI - Delayed loss of ETB receptor-mediated vasorelaxation after cold lesion of the rat parietal cortex. AB - The aim of this study was to investigate the involvement of endothelins (ET) in brain injury. The effect of ET was studied in the isolated basilar artery (BA) taken from control, sham-operated, and cold-lesioned rats. Cold lesion was induced by application of a precooled (-78 degrees C) copper cylinder (outer diameter 5 mm) for 60 seconds to the intact dura over the parietal cortex. After precontraction with prostaglandin (PG) F2alpha, ET-3 (10(-10) to 10(-8) mol/L) dilated BA with a pD2 (negative log of the half-maximal concentration) of 9.06+/ 0.031 (mean +/- SD) and a maximal effect (Emax) of 1.64+/-1.0 mN at 3 x 10(-9) mol/L in sham-operated animals. This dilation was reduced 24 and 48 hours after cold lesion by 33% and 73%, respectively, at 3 x 10(-9) mol/L. The effects of acetylcholine (10(-8) to 10(-4) mol/L) and sodium nitroprusside (10(-3) mol/L) were unaltered. Activation of the ETB receptor in thoracic aorta by the specific agonist IRL 1620 also resulted in a reduced dilation (51% by 48 hours after cold lesion). Reverse transcriptase-polymerase chain reaction of the BA showed unaltered expression of mRNA for the ETB receptor after cold lesion whereas ETB immunoreactivity in BA and in its intraparenchymal arteries was reduced at 24 and 48 hours. In contrast to the reduction of ET-3-induced dilation, the constrictor effects of ET-1 and ET-3 were retained after cold lesion. Endothelin-1 (10(-12) to 10(-6) mol/L) dose-dependently contracted segments of untreated control BA segments under resting conditions with a pD2 of 8.03+/-0.22 and an Emax of 6.35+/ 0.70 mN. Further evidence that the constrictor ability of BA was not influenced by cold lesion is given by the unaltered response to 124 mmol/L K+ and 10(-6) mol/L serotonin. We conclude that the ETB receptor of BA after cold lesion is downregulated specifically, apparently at the posttranscriptional level. Because the ETB-mediated dilation in thoracic aorta was also reduced, downregulation of the ETB receptor apparently is not restricted to cerebral arteries. The nitric oxide-cyclic guanosine monophosphate system in BA is, however, intact. PMID- 9850149 TI - An adiabatic approximation to the tissue homogeneity model for water exchange in the brain: I. Theoretical derivation. AB - Using the adiabatic approximation, which assumes that the tracer concentration in parenchymal tissue changes slowly relative to that in capillaries, we derived a time-domain, closed-form solution of the tissue homogeneity model. This solution, which is called the adiabatic solution, is similar in form to those of two compartment models. Owing to its simplicity, the adiabatic solution can be used in CBF experiments in which kinetic data with only limited time resolution or signal-to-noise ratio, or both, are obtained. Using computer simulations, we investigated the accuracy and the precision of the parameters in the adiabatic solution for values that reflect 2H-labeled water (D2O) clearance from the brain (see Part II). It was determined that of the three model parameters, (1) the vascular volume (Vi), (2) the product of extraction fraction and blood flow (EF), and (3) the clearance rate constant (kadb), only the last one could be determined accurately, and therefore CBF must be determined from this parameter only. From the error analysis of the adiabatic solution, it was concluded that for the D2O clearance experiments described in Part II, the coefficient of variation of CBF was approximately 7% in gray matter and 22% in white matter. PMID- 9850150 TI - An adiabatic approximation to the tissue homogeneity model for water exchange in the brain: II. Experimental validation. AB - A frequently reported limitation to using water as a tracer for measuring CBF has been the dependence of the CBF estimate on the experimental time (referred to as the falling flow phenomenon, FFP). To eliminate the FFP, we have developed the adiabatic solution of the tissue homogeneity model to replace the solution of the single-compartment Kety model. In Part I, the derivation of the adiabatic solution was presented. In this second part, the adiabatic solution was applied to measure CBF in rabbits using nuclear magnetic resonance spectroscopy and the tracer deuterium oxide. It was shown that the FFP, observable when the 2H clearance data were analyzed with the Kety equation, was significantly reduced when the same data were analyzed with the adiabatic solution of the tissue homogeneity model. By concurrently measuring CBF with radioactive microspheres, it was determined that the CBF estimates from the adiabatic solution were accurate for true blood flow values less than 60 mL x 100 g(-1) x min(-1). Above this value the CBF estimate was progressively underestimated, which was attributed to the diffusion limitation of water in the brain. PMID- 9850151 TI - Impact of aging on innate immunity. AB - Immune responses in higher organisms are triggered by the recognition of a limited diversity of microbiological products by cells of the innate or "natural" immune system. As a result, in addition to the direct protective effect of natural immunity, antigen-presenting cells, particularly dendritic cells, are activated to process and present an enormous number of peptide antigens to the T lymphocytes of the adaptive immune system. These, together with the B lymphocytes, then mediate specific immune responses and maintain acquired immunological memory. The aging immune system is less well able to cope with infectious disease than the youthful immune system; this review will briefly consider what is known of the age-associated alterations in innate immunity, and how these may also impact on adaptive immunity. PMID- 9850152 TI - ISCOMs: an adjuvant with multiple functions. AB - Aluminum salts are currently the only widely used adjuvant for human vaccines. Over the past 10-15 years, a large research effort has attempted to find novel adjuvants with ability to induce a broad range of immune responses, including cell-mediated immunity. The immunostimulating complex or ISCOM is one adjuvant with multiple adjuvant properties. ISCOMs are open cage-like complexes typically with a diameter of about 40 nm that are built up by cholesterol, lipid, immunogen, and saponins from the bark of the tree Quillaia saponaria Molina. ISCOMs have been demonstrated to promote antibody responses and induce T helper cell as well as cytotoxic T lymphocyte responses in a variety of experimental animal models, and have now progressed to phase I and II human trials. This review describes recent developments in the understanding of the structure, composition, and preparation of ISCOMs and will cover important aspects of the understanding of the adjuvant functions of ISCOMs and how they act on the immune system. PMID- 9850153 TI - Glucocorticoids protect against suppression of T cell responses in a murine model of acute ethanol exposure and thermal injury by regulating IL-6. AB - Previous reports by this laboratory demonstrated that acute alcohol exposure combined with a 15% body surface area dorsal scald injury results in significant reductions in delayed-type hypersensitivity (DTH) and splenocyte proliferative responses compared to either insult alone. Previous studies by this lab have also shown that these defects are mediated, in part, by increased production of interleukin-6 (IL-6). Because both alcohol exposure and thermal injury are known to modulate glucocorticoid (CORT) levels, and CORT regulates IL-6 gene expression, the relationship between circulating CORT and IL-6 production in burn + ethanol mice was examined. At 24 and 48 h post-burn, a positive correlation existed between circulating CORT levels and measurements of cellular immune function. Administration of exogenous CORT to burn + ethanol-treated mice resulted in significant restoration (to 60% of control) of DTH and splenocyte proliferative responses. This restoration was concomitant with a down-regulation of circulating and macrophage-derived IL-6. The specificity of CORT in modulating these responses was tested by assessing cellular immune function and IL-6 levels after glucocorticoid receptor blockade with RU486. Taken together, these data strongly suggest that under normal circumstances CORT protects burned mice from severe immune dysfunction, a protection that is not afforded to burn + ethanol treated mice. Furthermore, the immune dysfunction observed in burn + ethanol mice may be due to a lack of glucocorticoid attenuation of IL-6. PMID- 9850154 TI - Extrathymic T cell differentiation in vitro from human CD34+ stem cells. AB - Although it is well established that T cells are derived from CD34+ stem cells in vivo, and that T cells can develop in the absence of a functioning thymus, it has not proven possible thus far to generate human T cells in vitro from CD34+ cells in the absence of any thymic influence. We now present a limiting dilution cloning culture system that supports the differentiation of highly purified human CD34+ cells to CD3+ T cells in vitro in the complete absence of any thymic components. The culture system features the use of a serum-free medium supplemented with a cocktail of cytokines including flt-3 ligand, interleukin-3 (IL-3), stem cell factor (SCF), and IL-2. CD4+ T cell clones capable of mitogen stimulated proliferation and response to IL-2, and expressing a varied TCR-Vbeta repertoire were obtained under these conditions. This culture system therefore supports human T lymphopoiesis in the absence of any thymic influence and may prove useful for the evaluation of extrathymic T cell differentiation in vitro. PMID- 9850155 TI - Thermal injury alters macrophage responses to prostaglandin E2: contribution to the enhancement of inducible nitric oxide synthase activity. AB - Prostaglandin E2 (PGE2) and macrophage (Mphi)-derived reactive nitrogen intermediates (RNI) have been implicated in T cell dysfunction after thermal injury. Normally, Mphi inducible nitric oxide synthase (iNOS) activity can be regulated by PGE2, however, it is unknown whether PGE2 modulates Mphi iNOS activity after thermal injury. Splenic Mphi isolated from mice 7 days after thermal injury produced higher levels of RNI than Mphi from sham mice when stimulated with lipopolysaccharide (LPS) or interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) in combination. PGE2, when added concurrently with LPS, suppressed RNI production by Mphi from sham mice, whereas Mphi from injured mice were unaffected. When Mphi were pretreated with PGE2 before LPS, RNI production was suppressed in both populations. RNI production in response to IFN-gamma or IFN-gamma and TNF-alpha in combination was enhanced by PGE2 in both populations, however, the effect was markedly greater in Mphi from injured mice. The PGE2-mediated changes in RNI production were paralleled by similar changes in iNOS protein expression, suggesting that the effect of PGE2 was at the level of enzyme expression rather than activity. Dibutryl cAMP induced similar effects as PGE2, suggesting the response to PGE2 after thermal injury is independent of potential changes in PGE2-induced adenylate cyclase activity and is cAMP-mediated. The results indicate that Mphi from burned mice display an altered sensitivity to PGE2, resulting in enhanced iNOS activity. Thus, PGE2, which is elevated after thermal injury and can directly suppress T cell function, may also contribute to immune dysfunction through the enhancement of Mphi iNOS activity. PMID- 9850156 TI - Nordihydroguaiaretic acid blocks secretory and endocytic pathways in human dendritic cells. AB - Nordihydroguaiaretic acid (NDGA), an antioxidant and inhibitor of the lipoxygenase arm of the arachidonic acid metabolism, was recently demonstrated to inhibit transport of secretory proteins to the Golgi complex. We have investigated the effects of NDGA on the secretory and endocytic activity of cultured human blood dendritic cells (DC). Treatment with NDGA strongly diminished cytokine secretion by DC. Moreover, NDGA reduced in a dose- and time dependent fashion fluid phase as well as receptor-mediated endocytosis in DC. Zileuton and MK-886, specific inhibitors of 5-lipoxygenase and 5-lipoxygenase activating protein, respectively, had no effect. Likewise, N-acetyl-L-cysteine, a thiol antioxidant precursor of glutathione, did not affect DC function. Finally, serum remarkably protected the cells from the inhibitory effects of NDGA. Our data demonstrate that NDGA not only disrupts vesicular transport along the secretory route but is also a potent inhibitor of the endocytic pathways in human DC and that NDGA has inhibitory properties different from those described. PMID- 9850157 TI - Beta 1 integrin-dependent binding of Jurkat cells to fibronectin is regulated by a serine-threonine phosphatase. AB - We investigated the effects of signaling molecule inhibitors on the expression and function of beta1 integrins in Jurkat cells. Jurkat cells expressed alpha4beta1 and alpha5beta1, with significant levels of constitutively activated beta1 integrins as assessed by labeling with mAb 15/7 that distinguishes between activation states. Adhesion to fibronectin (Fn) was mediated equally through alpha4 and alpha5 subunits, and was potentiated by the beta1 integrin activating mAb 8A2. Fn adhesion was decreased by okadaic acid through effects on both alpha4beta1, and alpha5beta1. Tyrphostin A23 also decreased adhesion but was less potent. Neither inhibitor had any effect on the surface expression of total or activated beta1 integrins. The effect of tyrphostin was completely reversed by 8A2; the effect of okadaic acid was only partially reversed. Using Calyculin A, we determined that Jurkat adhesion to Fn was regulated via protein phosphatase 1, independent of the levels of integrins or integrin activation epitopes. Activation of Jurkat cells with a CD3-stimulating mAb enhanced adhesion to Fn and was partially blocked by okadaic acid. These data demonstrate different regulatory pathways for constitutive versus activation-dependent adhesion via beta1 integrins, and implicate both tyrosine kinases and serine-threonine phosphatases in integrin function. PMID- 9850158 TI - Membrane depolarization and depletion of intracellular calcium stores are associated with delay of apoptosis in human neutrophils. AB - Apoptosis occurs rapidly in human polymorphonuclear leukocytes (PMN) after exposure to 1 mM cycloheximide (CHX). We examined whether this form of stimulated apoptosis altered either resting cytosolic free Ca2+ concentrations ([free Ca]) or membrane potential (psi) in PMN and found no significant effects. However, manipulation of either PMN intracellular Ca2+ stores or psi was found to delay CHX-induced apoptosis. Depletion of PMN intracellular Ca2+ stores with thapsigargin caused membrane depolarization and significantly delayed CHX-induced apoptosis based on both morphological and annexin-V-fluorescein isothiocyanate binding criteria. Short-term suspension (4 h) of PMN in Ca2+-free buffer depleted internal Ca2+ stores, induced membrane depolarization at 2.5 h, and delayed spontaneous (24 h) apoptosis but had no effect on CHX-induced apoptosis. Rapid membrane depolarization with 150 mM KCl buffer significantly delayed CHX-induced apoptosis, suggesting that depolarization rather than Ca2+ stores depletion was the crucial event. Timing experiments revealed that depolarization within 12 min of CHX exposure significantly delayed apoptosis. Collectively, these observations suggest an early psi-sensitive step in the apoptosis pathway initiated by CHX. CHX exposure alone does not alter either resting PMN [free Ca] or psi; accompanying depolarization of plasma membrane (either electrochemically or via depletion of internal Ca2+ stores) delays CHX-induced apoptosis in a time dependent manner. PMID- 9850159 TI - Expression of a structural domain of the beta 2 subunit essential for alpha M beta 2 ligand recognition. AB - The beta2 leukocyte integrins comprise a group of closely related adhesion receptors that mediate critical events during normal and inflammatory immune responses. Central to the understanding of beta2 integrin function is the basis of ligand recognition. Results from our laboratory and others indicate the presence of multiple ligand contact points in both the alpha and beta subunit. As an approach to identify and characterize regulatory domains of the beta2 subunit, we have generated two different subdomains of the beta2 subunit for expression on the surface of mammalian cells through a phosphatidyl-inositol glycan anchor. The first subdomain contains the putative beta2 MIDAS motif implicated in ligand binding [beta2(LB)], whereas the second beta2 subdomain contains the cysteine rich region [beta2(CR)]. Cells expressing alphaM and beta2 constructs singly or cotransfected transiently in COS-7 cells were tested for the ability to bind to immobilized iC3b. Cells bearing the recombinant alphaMbeta2(LB) were capable of adhering to iC3b in a manner similar to that observed with the complete alphaMbeta2 heterodimer. In contrast, cells expressing alphaMbeta2(CR) failed to adhere to immobilized iC3b. Moreover, cells bearing singly transfected alpha or beta chains alone failed to adhere to immobilized iC3b. These results indicate that along with alphaM, the beta2(LB) subdomain contains the sufficient components within the beta2 subunit essential for ligand recognition. These findings support the hypothesis that the beta2 subunit cooperates with site(s) within the alphaM subunit in a receptor/cation/ligand complex resulting in high affinity ligand interaction. PMID- 9850160 TI - Nucleosomes inhibit phagocytosis of apoptotic thymocytes by peritoneal macrophages from MRL+/+ lupus-prone mice. AB - The nucleosome, the basic structure of chromatin and normal product of cell apoptosis, plays a pivotal role both in the induction and the pathogenesis of systemic lupus erythematosus (SLE). Nucleosomes have been found to circulate at high levels in patients with SLE and apoptosis of lymphoid cells is increased during human and murine lupus. In this study, we examined the presence of possible defects in clearance mechanisms of apoptotic cells in murine lupus, and questioned further whether nucleosomes could compromise this phagocytic process. There did not appear to be any intrinsic functional defect of macrophages from young MRL+/+ lupus-prone mice to recognize and phagocytose apoptotic thymocytes. Nucleosomes, as a mimic of increased cell apoptotsis in vivo, induced a strong, dose-dependent, inhibition of phagocytosis of apoptotic thymocytes by young, pre autoimmune, macrophages of MRL+/+ mice, whereas macrophages of non-autoimmune C3H mice only exhibited a trend to inhibition. The nucleosome-elicited inhibitory effect persisted during the development of the autoimmune response and appeared to be specific for the molecular mechanisms involved in macrophage phagocytosis of apoptotic cells. Our data suggest that nucleosome elicited inhibition of phagocytosis of apoptotic cells by MRL+/+ macrophages before the onset of the autoimmune response contribute, in a positive loop, to sustain and/or augment the levels of circulating (and potentially immunogenic) nucleosomes in lupus. PMID- 9850161 TI - During human thymic development, beta 1 integrins regulate adhesion, motility, and the outcome of RHAMM/hyaluronan engagement. AB - During human thymic differentiation, interactions between fibronectin (Fn)/beta1 integrins and hyaluronan (HA)/RHAMM control motility and Fn/beta1 integrins mediate spontaneous Fn-dependent adhesion. Multinegative (MN, CD3-4-8-) thymocytes exhibit strong spontaneous adherence to Fn (75%) that was efficiently inhibited by anti-alpha5beta1 and only weakly inhibited by anti-alpha4beta1. The relatively weak adherence of unfractionated thymocytes to Fn required both alpha4beta1 and alpha5beta1. Video time-lapse microscopy indicates that a subset of thymocytes also undergo spontaneous Fn-dependent motility mediated by alpha5beta1, alpha4beta1, and the HA-receptor RHAMM, but not by CD44. The loss of motility after hyaluronidase treatment of thymocytes indicated that motility is strongly dependent on HA. Of motile cells, 55% were DP, 19% were DN, and 24% were CD4+SP, but only 1% were CD8+SP. Overall, for MN thymocytes, beta1 integrin mediated Fn-adhesion, but after expression of CD4/CD8, beta1 integrins mediated Fn-dependent motility. Treatment with the activating anti-beta1 mAb QE.2E5 inhibited thymic motility and converted otherwise nonadherent thymocytes to an adherent state. High-avidity interactions via integrins appear to supercede the motogenicity of RHAMM and HA, suggesting that integrin avidity may regulate RHAMM. During thymic development, changes in adhesion or motility appear to be mediated by integrin avidity modulation. PMID- 9850162 TI - Expression of functional major histocompatibility complex class II molecules on HMC-1 human mast cells. AB - Mast cells hold a key position in the defensive mechanisms against exogenous intruders. In this study, we investigated whether human mast cells express functional major histocompatibility complex (MHC) class II molecules that can transduce endogenous signals and present staphylococcal enterotoxin A (SEA) to T cells. Similar to HMC-1 human mast cell line, umbilical cord blood-derived mast cells express HLA-DR, -DP and -DQ molecules on their surface. MHC class II molecules expressed on HMC-1 cells bind significantly the SEA (a natural MHC class II ligand), and their ligation with specific mAbs or with SEA, leads ultrastructural changes, suggesting their degranulation. Recognition of SEA-bound MHC class II molecules on HMC-1 mast cells by the T cell receptor of K25 cells, an SEA-specific murine T cell hybridoma, triggers significant IL-2 secretion by these T cell hybridomas. Hence, our data point out the expression of functional MHC class II molecules on human mast cells, reinforcing the implication of these cells in the defense mechanisms of acquired immunity. PMID- 9850163 TI - CD43 is associated with tyrosine kinase activity in human neutrophils. AB - CD43 appears to be involved in signal transduction, however, the mechanism of this function is unknown. Protein kinase activity was detected in neutrophils associated with CD43. Most of the protein kinase activity associated with these antigens was tyrosine kinase activity. The src family kinases lyn and hck were found to account for much of the associated tyrosine kinase activity. The data suggest that associated tyrosine kinase activity may play a role in signal transduction via CD43 to regulate other cell functions. PMID- 9850164 TI - Costimulatory molecules CD80 and CD86 in the rat; tissue distribution and expression by antigen-presenting cells. AB - The CD28-CD80/CD86 costimulatory pathway provides a critical signal for T cell activation. Only recently rat CD80 and CD86 have been cloned and monoclonal antibodies have been generated. In this study we examined the expression of these molecules in lymphoid tissue and on purified subsets of antigen-presenting cells (APC). The target tissue of cyclosporin A-induced autoimmunity, i.e. the skin and tongue, were also examined for expression of CD80 and CD86. Whereas CD80 was hardly detected in the lymphoid tissues, CD86 was clearly expressed by non lymphoid cells in the thymus, as well as in the secondary lymphoid organs. With respect to lymphoid cells, only germinal center B cells exhibited clear CD86 expression. Phenotypic analysis by flow cytometry revealed that only dendritic cells, both of thymic and splenic origin, expressed the full array of stimulatory molecules required for the proper activation of naive T cells. On development of cyclosporin A-induced autoimmunity, non-professional APC, i.e. epithelial cells, started to express MHC class II, but not the costimulatory ligands CD80 and CD86. However, CD86 staining was observed in the target tissue and was associated with Langerhans cells as well as infiltrating leukocytes. Altogether, our results show that also in the rat strong stimulatory capacity for primary immune responses is associated with the expression of the costimulatory ligands CD80 and CD86. As concluded from the in situ expression CD86 may be the predominant costimulatory ligand early in immune responses. PMID- 9850165 TI - CSF-1 regulation of Il6 gene expression by murine macrophages: a pivotal role for GM-CSF. AB - We test the hypothesis that the monocyte-macrophage colony-stimulating factor (CSF-1 or M-CSF) plays a major role in the inflammatory responses of Mphi by acting as a priming agent that heightens their responsiveness to secondary stimulation by other mediators. We previously reported that CSF-1 induced peritoneal Mphi (PMphi) to transcribe several genes including interleukin-6 (Il6) and granulocyte-macrophage colony-stimulating factor (Csfgm). It was reported that the Il6 and Csfgm genes were individually regulated by different pathways but it was not clear to what extent the two genes interacted during Mphi-mediated inflammatory responses. We now show that CSF-1 induces the release of bioactive GM-CSF from mouse resident PMphi. GM-CSF induces Il6 gene expression and synergizes with CSF-1 to induce the release of large amounts of IL-6. PMphi from C57BL/6J-Csfgm(null) mice were shown to release minimal IL-6 in response to CSF-1 and to express a much reduced response to the highly stimulatory combination of CSF-1 and lipopolysaccharide (LPS). Exogenous recombinant GM-CSF restored the IL 6 response of GM-CSF null PMphi to a great extent but not completely. As controls, three other recombinant proteins were tested but of these only tumor necrosis factor alpha (TNF-alpha) was shown to synergize with both CSF-1 and GM CSF. Using PMphi from mice deficient in the expression of the Il6 gene, it was shown that they released two- to threefold more GM-CSF in response to CSF-1 than their control counterparts. However, an exogenous supply of recombinant IL-6 had no effect on GM-CSF release. The data indicate that the pathways regulating Il6 gene expression are under the control of a complex network of cytokine interactions involving at least CSF-1, GM-CSF, and TNF-alpha, with the added possibility that IL-6 may exert modulatory activity within this network. PMID- 9850166 TI - Platelet-activating factor induces a concentration-dependent spectrum of functional responses in bovine neutrophils. AB - We characterized the dose response of bovine neutrophils to platelet-activating factor (PAF) with respect to the following functions: calcium flux and membrane potential changes, actin polymerization, degranulation, and the production and/or priming of the oxidative burst. PAF at very low concentrations (10(-10) and 10( 9) M) caused changes in intracellular calcium and membrane potential in bovine neutrophils, whereas moderate PAF concentrations (> or = 10(-7) M) resulted in increased actin polymerization. Degranulation responses to PAF were more complex: low concentrations (10(-9) M) caused secretory granule degranulation, moderate doses (> or = 10(-7) M) caused specific granule degranulation, whereas azurophil degranulation only occurred at high (10(-5) M) PAF concentrations. Treatment of bovine neutrophils with PAF at concentrations > or = 10(-7) M also caused up regulation of the adhesion molecules Mac-l and L-selectin. PAF stimulation resulted in a very weak [compared to phorbol myristate acetate (PMA)] oxidative burst in bovine neutrophils, and only at high (10(-6) M) concentrations. Unlike human neutrophils, bovine neutrophils were poorly primed by PAF treatment. Only high concentrations of PAF (10(-5) M) caused an increased rate of PMA-stimulated superoxide production, although lower doses of PAF did reduce the lag time preceding the PMA-induced oxidative burst. The overall pattern that can be inferred is that lower concentrations of PAF promote neutrophil sensitivity and interaction by selective degranulation, up-regulation of adhesion molecules, and increased actin polymerization. In contrast, higher PAF concentrations can promote, albeit weakly, more direct bactericidal responses, such as the release of reactive oxygen species and granule enzymes. The ability of PAF to modulate a graded response in bovine neutrophils would allow the cell to respond proportionally to the severity of a stimulus. PMID- 9850167 TI - Signaling in neuropeptide-induced migration of human eosinophils. AB - At inflammatory sites, leukocytes may confront multiple, competing chemoattractive signals. We compared the chemotactic potencies of several sensory neuropeptides with regard to signal transduction pathways in eosinophils. Eosinophils were enriched using magnetic cell sorting and migration was assayed in a Boyden microchemotaxis chamber. We found stimulatory effects of substance P, calcitonin gene-related peptide (CGRP), secretoneurin, vasoactive intestinal peptide (VIP), and secretin on eosinophil migration. Actions of VIP are predominantly mediated via VIP receptor type I. Migration toward secretoneurin, VIP, and secretin was blocked by a phosphodiesterase inhibitor, which, in contrast failed to affect substance P- and CGRP-induced eosinophil chemotaxis. Wortmannin blunted the migratory responses induced by all neuropeptides tested and substance P-induced effects on eosinophils were tyrphostin-23-sensitive. We conclude that substance P, CGRP, secretoneurin, and VIP/secretin stimulate eosinophil migration involving wortmannin-sensitive enzymes. Moreover, secretoneurin and VIP/secretin require additional activation of phosphodiesterases to stimulate eosinophil migration. PMID- 9850168 TI - Bacterial phagocytosis activates extracellular signal-regulated kinase and p38 mitogen-activated protein kinase cascades in human neutrophils. AB - The hypothesis that bacterial phagocytosis by human polymorphonuclear neutrophils (PMNs) stimulates MAPK cascades that regulate respiratory burst activation was tested. Extracellular response kinase (ERK) and p38 kinase, but not c-Jun NH2 terminal kinase, activities were increased within 5 min of phagocytosis of plasma opsonized Staphylococcus aureus (S-SA), reached maximum at 20-30 min, and remained elevated through 60 min. The role of Fcy receptors was examined using gamma globulin-opsonized SA (IgG-SA), whereas CR3 receptors were activated by particulate beta-glucan. IgG-SA stimulated a maximal ERK activity at 30 min, whereas p38 activity was maximal at 5 min. Beta-glucan stimulated maximal ERK activity at 5 min and maximal p38 activity at 2 min. Non-opsonized bacteria were ingested at 10% of the level of S-SA and stimulated a minimal increase in ERK and p38 activity at 60 min. S-SA stimulation of ERK was inhibited by wortmannin, LY294002, and genistein, but not calphostin C; whereas p38 stimulation was inhibited by calphostin C and genistein, but not wortmannin and LY294002. Simultaneous measurement of phagocytosis and H2O2 production by flow cytometry was used to assess the role of ERKs and p38 kinase in phagocytosis. The MEK inhibitor PD098059 had no significant effect on phagocytosis or H2O2 production. The p38 kinase inhibitor SB203580 significantly attenuated H2O2 production, whereas phagocytosis was unaffected. In conclusion, bacterial phagocytosis stimulates ERK and p38 activation by distinct signal transduction pathways. Phagocytosis-stimulated p38 kinase activity is necessary for optimal H2O2 production. PMID- 9850169 TI - Evaluation of the expression of human CAP18 gene during neutrophil maturation in the bone marrow. AB - To understand the gene expression of CAP18 (18-kDa cationic antibacterial protein), a member of cathelicidins, we evaluated mRNA and protein expression of CAP18 using human bone marrow cells and mature neutrophils. Northern blot analysis revealed that CAP18 mRNA was expressed more abundantly in bone marrow cells than mature neutrophils, whereas Western blot analysis indicated that CAP18 protein was more abundant in mature neutrophils than bone marrow cells. Consistent with this, in situ hybridization using bone marrow cells demonstrated that the expression of CAP18 mRNA was neutrophil lineage-specific and was observed primarily in myelocytes (>95%) with limited expression in more immature cells (promyelocytes) and mature cells (metamyelocytes, band cells, and segmented neutrophils). Furthermore, immunohistochemical study indicated that, coincident with the increase of CAP18 mRNA levels, CAP18-positive cells increased markedly at myelocyte stage, and the increased levels remained almost constant (>95%) in metamyelocytes, band cells, and segmented neutrophils, although the mRNA levels were remarkably reduced in these cells. Together these observations indicate that CAP18 gene transcription likely occurs lineage- and stage-specifically at the myelocyte stage of neutrophil maturation in the bone marrow and results in the synthesis and cytoplasmic accumulation of CAP18, which is present in the subsequent stages of neutrophil maturation. PMID- 9850170 TI - The World Health Organization/International Society of Urological Pathology consensus classification of urothelial (transitional cell) neoplasms of the urinary bladder. Bladder Consensus Conference Committee. AB - This WHO/ISUP system is an attempt to develop as broad a consensus as possible in the classification of urothelial neoplasms, building upon earlier works and classification systems. It is meant to serve as a springboard for future studies that will help refine this classification, thus enabling us to provide better correlation of these lesions with their biologic behavior using uniform terminology. PMID- 9850171 TI - Primary ovarian mucinous cystadenocarcinomas: a clinicopathologic study of 49 cases with long-term follow-up. AB - Prognostic data for ovarian mucinous carcinoma are limited and difficult to interpret because of differing diagnostic criteria and inclusion of secondary tumors. To better characterize these neoplasms, 49 primary ovarian mucinous tumors diagnosed as carcinoma by the Hart and Norris criteria and staged by the FIGO system were studied. Forty-four tumors (90%) were stage I, four were stage III and one was unstaged. Sixteen tumors (33%) were classified as intraglandular ("noninvasive") carcinoma; all were stage I and all patients were alive without tumor after 4-216 months (mean, 74 months); two patients had received adjuvant chemotherapy. Stromal invasion was present in the remaining 33 cases (67%), including 19 tumors with extensive invasion and 14 with one or more discrete foci of microinvasion (each focus < or = 1 mm). The microinvasive tumors were reclassified into intraglandular carcinoma with microinvasion (nine cases) and borderline (low malignant potential) tumor with microinvasion (five cases). All microinvasive tumors were stage I and none recurred after postoperative intervals of 9-176 months (mean, 71 months) for the microinvasive carcinomas and 33-117 months (mean, 60 months) for the microinvasive borderline tumors; only 1 of the 14 patients received adjuvant chemotherapy. All 19 extensively invasive carcinomas also had intraglandular carcinoma. Fourteen were stage I, four were stage III, and one was unstaged. Eleven (79%) of the stage I patients were alive without tumor after 10-220 months (mean, 110 months), including six who received chemotherapy; one was dead without tumor and two developed progressive disease (one had received adjuvant chemotherapy). The four extensively invasive stage III carcinomas were fatal after 1-59 months. The unstaged patient received adjuvant chemotherapy and was alive without recurrence at 98 months. Conclusions of this study are as follows: (1) primary mucinous carcinomas are very uncommon tumors, after rigorous exclusion of metastatic carcinomas and tumors associated with pseudomyxoma peritonei; (2) bilaterality is not a feature of primary mucinous carcinomas; (3) FIGO stage is the single most important prognostic factor, with stage I carcinomas having a very favorable prognosis; (4) stage I carcinomas that metastasize have extensive stromal invasion; (5) extensive stromal invasion is found only in tumors with a component of intraglandular carcinoma; (6) high-stage carcinomas invariably contain extensively invasive carcinoma and have a very poor prognosis; and (7) stromal microinvasion with individual foci not exceeding 1 mm does not appear to be an adverse factor in either carcinomas or borderline tumors of stage I. PMID- 9850172 TI - p53 immunostaining as a significant adjunct diagnostic method for uterine surface carcinoma: precursor of uterine papillary serous carcinoma. AB - Uterine papillary serous carcinoma (UPSC) is a biologically aggressive carcinoma that causes a disproportionate number of endometrial cancer deaths because of its dismal clinical outcome. Although the precursor lesion of UPSC has been suggested both morphologically and molecularly, diagnosis continues to represent a challenge to surgical pathologists, particularly in biopsy specimens, largely in part because of its multiple histologic patterns and many benign morphologic mimics. In this study, we used p53 immunohistochemical staining as an adjunct test to correctly identify six cases of uterine surface carcinoma (USC) prospectively and three cases retrospectively. Both sensitivity and specificity for this immunostaining method approached 100% when the cutoff score of p53 overexpression was 7 or higher. The precision estimated by receiving operating characteristic curve was 100%, indicating that the diagnostic value of the score for p53 overexpression was very high. p53 immunohistochemical staining was considered a significant adjunct diagnostic method for the probable precursor lesion of UPSC. The probable precursor lesion of UPSC, previously referred to as endometrial intraepithelial carcinoma or endometrial carcinoma in situ, appears to represent the early phase of UPSC. However, unlike its names would suggest, this lesion is often multicentric and behaves in a more aggressive fashion than regular in situ carcinomas. For this reason, we prefer the term uterine surface carcinoma, a term that is more descriptive and less restrictive, to emphasize the unique aggressive nature of the UPSC precursor lesion. The reason we postulate using the term uterine surface carcinoma rather than endometrial intraepithelial carcinoma or endometrial carcinoma in situ is that the latter terms would seem define a neoplastic process confined to the endometrial epithelium without potential for metastasis. In reality, the precursor lesion of UPSC has a tendency to stromal and vascular space involvement as seen by the presence of stromal and vascular invasion in one of the prospectively identified USC cases. Therefore, the term uterine surface carcinoma is selected to alert clinicians that this early carcinoma has features of carcinoma in situ, but still carries a potential for metastasis. PMID- 9850173 TI - Thymic carcinoma arising in thymoma is associated with alterations in immunohistochemical profile. AB - Thymic carcinoma is an uncommon tumor. Most cases appear to arise de novo, but in rare instances they can arise in thymomas. We report the clinicopathologic features and immunohistochemical profile of five cases of thymic carcinoma accompanied by a component of thymoma. Immunohistochemical studies were performed with the avidin-biotin-peroxidase complex method using monoclonal antibodies to p53(DO7), CD99(O13), epithelial membrane antigen, CD5(NCL-CD5-4C7), vimentin (V9), and cytokeratins 7, 8, 18, and 19. The patients consisted of three men and two women with a median age of 57 years. One patient had myasthenia gravis, and the other four presented with chest symptoms. One patient had concurrent adenocarcinoma of the lung with metastasis. Four of the patients died within 15 months. The thymomas consisted of two large polygonal cell thymomas, two squamoid thymomas, and one spindle cell thymoma. The malignant components included two undifferentiated carcinomas, one spindle cell carcinoma, one squamous cell carcinoma, and one clear cell carcinoma with squamous differentiation. There was no correlation between the histologic types of the thymoma and the thymic carcinoma. In three cases, excluding the two squamoid thymomas, the thymic carcinomas occurred in the necrotic areas of the thymoma. They showed upregulated expression of epithelial membrane antigen and cytokeratins 7, 8, 18, and 19, similar to the so-called "interface phenomenon" described in the invasion front of other types of carcinoma. Increased p53 protein expression was observed in all five carcinomas, and there was loss of CD99+ immature T lymphocytes. Among the thymic carcinomas, only the squamous component of the clear-cell carcinoma stained for CD5, a marker commonly expressed in thymic carcinomas. Paradoxically, a squamoid thymoma, but not its associated spindle cell carcinoma, expressed CD5, suggesting the acquisition of an "aggressive" phenotype by the squamoid thymoma, but with loss of the marker on malignant transformation. One undifferentiated carcinoma acquired vimentin immunoreactivity, whereas four other carcinomas and all five thymomas were negative. In conclusion, thymic carcinoma can arise in any histologic type of thymoma, including spindle cell thymoma, which is generally regarded as a benign neoplasm. The prognosis appears to be poor. Tumor necrosis in a thymoma should alert the pathologist to search for malignant change. The malignant change is commonly associated with increased expression of epithelial membrane antigen, cytokeratin subtypes, or p53 protein, and loss of CD99+ immature T lymphocytes, and is occasionally associated with a change in the expression of CD5 or vimentin. PMID- 9850174 TI - "Rhabdoid" meningioma: an aggressive variant. AB - It is has been suggested that rhabdoid morphology is associated with a poor prognosis, regardless of tumor histogenesis. We report a series of 15 meningiomas with rhabdoid features. Nine patients had undergone multiple resections. In six, the rhabdoid component was histologically apparent only in recurrences. Rhabdoid morphology was defined as sheets of loosely cohesive cells with eccentric nuclei and hyaline, paranuclear inclusions. Ultrastructurally, the latter consisted of whorls of intermediate filaments often entrapping lysosomes or other organelles. Meningothelial features included whorl formation and nuclear pseudoinclusions, immunohistochemical coexpression of vimentin and epithelial membrane antigen, and the ultrastructural finding of interdigitating cell membranes and intercellular junctions. At the histologic level, a conventional meningioma component was noted in most tumors; only four lesions were entirely rhabdoid. Histologic malignancy (brain invasion or anaplasia) was observed in nine cases, another two tumors being considered malignant on the basis of extracranial metastasis. In the majority, increased cell proliferation was evidenced by a high mitotic rate or MIB-1 LI. At last follow-up, 13 patients (87%) had experienced at least one recurrence and 8 (53%) were dead of disease. Median time to death was 5.8 years after initial surgery and 3.1 years after the first appearance of rhabdoid morphology. Our findings corroborate those from a smaller series recently reported by Kepes et al. on the same entity (Kepes JJ, Moral LA, Wilkinson SB, Abdullah A, Llena JF. Rhabdoid transformation of tumor cells in meningiomas: A histologic indication of increased proliferative activity. Report of four cases. Am J Surg Pathol 1998;22:231-8). They further suggest that rhabdoid meningiomas are highly aggressive tumors and that the rhabdoid phenotype represents a marker of malignant transformation in meningiomas. PMID- 9850175 TI - Cancer volume of lymph node metastasis predicts progression in prostate cancer. AB - Clinical outcome is variable in prostate cancer patients with regional lymph node metastasis. We studied 269 patients who had regional lymph node metastasis at the time of radical retropubic prostatectomy and bilateral pelvic lymphadenectomy at the Mayo Clinic between January 1987 and December 1992. Two hundred fifty-three (94%) patients received androgen deprivation therapy within 90 days of radical prostatectomy. Patients ranged in age from 47 to 79 years (median, 67 years). Median follow-up was 6.1 years (range, 0.3-10.5 years). Nodal cancer volume (size) was measured by the grid-counting method. Cox proportional hazards models were used to determine the impact of numerous clinical and pathologic findings on systemic progression-free survival. Systemic progression was defined as the presence of distant metastasis documented by biopsies or radiographic examinations (abdominal computerized tomography, plain radiographs, or bone scan). Five-year progression-free survival was 90%. In predicting systemic progression using Cox multivariate analysis, only nodal cancer volume added significantly to the model containing the primary cancer variables (Gleason score, cancer volume, and DNA ploidy). The relative hazard rate for a doubling in nodal cancer volume was 1.6 (95% confidence interval, 1.3 to 2.0; p < 0.0001). Spearman rank analysis showed a correlation between nodal cancer volume and Gleason score of the primary cancer, the number of positive nodes, the aggregate length of metastases, and the largest nodal cancer diameter (correlation efficient = 0.37, 0.63, 0.96, and 0.95, respectively). Our data indicate that nodal cancer volume was the most significant nodal determinant of progression to distant metastasis in lymph node-positive prostate cancer patients. We recommend that the diameter of the largest metastasis be evaluated in patients with metastases, because this is a more powerful predictor of patient outcome than current methods, which recommend mere counting of the number of positive nodes. PMID- 9850176 TI - Atypical and malignant solitary fibrous tumors in extrathoracic locations: evidence of their comparability to intra-thoracic tumors. AB - Solitary fibrous tumor (SFT), first described as a pleural lesion, has been reported at numerous extrathoracic sites over the past 10 years. About 10% to 15% of intrathoracic SFTs are histologically or clinically malignant, but such cases have very rarely been described at other locations. Among 92 cases of extrathoracic SFT in our files, we identified 10 that either had recurred (2 cases) or had a least one atypical histologic feature (8 cases). The ten tumors occurred in five men and five women, 32 to 81 years old (median 56), measured 1.9 cm to 20 cm (median 11.5 cm), and were located in the abdomen/pelvis (4 cases), retroperitoneum (3 cases), groin, trunk, and upper arm. Nuclear atypia (8 cases), markedly increased cellularity (6 cases), areas of necrosis (4 cases), and greater than 4 mitoses/10 HPFs (3 cases) were seen in addition to the typical histologic features of SFT. Six tumors had at least two of these atypical histologic features. Nine cases were positive for CD34, six were positive for O 13, and one was focally positive for smooth muscle actin. Eight were excised completely. Subsequent follow-up revealed tumor relapse in eight cases (follow up 6-180 months, median 24). Four patients had local recurrence at 12 to 168 months. Distant metastasis developed at 1 to 6 years in five cases with spread to lung (2 cases), liver (4 cases), and bone. Metastasis or local recurrence developed within 2 years in five patients. To date, no patient has died of their tumor. These findings demonstrate that nuclear atypia, hypercellularity, greater than 4 mitoses/10 HPFs, and necrosis may be seen in up to 10% of extrathoracic SFTs, and are associated with, but are not by themselves predictive of, aggressive clinical behavior. In addition, our findings confirm that the behavior of extrathoracic SFTs is unpredictable, entirely comparable to that of their better known pleural counterparts, and confirm that patients with SFTs in all locations require careful, long-term follow up. It is probably unwise to regard any such lesion as definitely benign. PMID- 9850177 TI - Follicular and Hurthle cell carcinomas of the thyroid: a comparative study. AB - University of Texas M. D. Anderson Cancer Center cases filed as Hurthle cell and follicular carcinoma were reviewed. Requirements for including a case in the study were that the diagnosis of Hurthle cell or follicular carcinoma be confirmed, that histologic material and clinical information be adequate, and that there be at least 9 years of follow-up. The study group included 18 cases of Hurthle cell carcinoma and 33 cases of follicular carcinoma. Ten of the Hurthle cell carcinomas had extrathyroid invasion, three had intrathyroid invasion, and five were encapsulated (i.e., they had intracapsular invasion only). In the follicular carcinoma group, 5 tumors had extrathyroid invasion, 14 had intrathyroid invasion, and 14 were encapsulated. When the cases were stratified according to extent of invasion in this manner, there was no statistically significant difference in rate of local recurrence, rate of metastasis (either regional lymph node or distant), or patient survival between Hurthle cell carcinoma and follicular carcinoma. Other variables including patient age and sex, treatment differences, tumor size, vascular invasion, predominant growth pattern (follicular versus solid-trabecular), nuclear size and pleomorphism, mitotic rate, and tumor necrosis did not provide significant additional prognostic information. Metastases of both Hurthle cell and follicular carcinoma were mostly distant and predominantly involved bone and lung. Behavioral differences between Hurthle cell and follicular carcinoma that were not statistically significant included a higher rate of local recurrence in Hurthle cell carcinoma with intrathyroid invasion, more frequent occurrence of regional lymph node metastasis in Hurthle cell carcinoma with extrathyroid invasion, and absence of distant metastasis and death caused by tumor in encapsulated Hurthle cell carcinoma. Five follicular carcinomas and one Hurthle cell carcinoma appeared to have arisen within an adenoma. PMID- 9850178 TI - Columnar alteration with prominent apical snouts and secretions: a spectrum of changes frequently present in breast biopsies performed for microcalcifications. AB - We have noted in breast biopsies performed for microcalcifications a spectrum of lesions in the terminal duct lobular unit (TDLU) characterized by columnar epithelial cells with prominent apical cytoplasmic snouts, intraluminal secretions, and varying degrees of nuclear atypia and architectural complexity. The appearance of some of these lesions is worrisome, but diagnostic difficulties arise because the histologic features do not fulfill established criteria for the diagnosis of atypical ductal hyperplasia or ductal carcinoma in situ (DCIS). We have termed such lesions columnar alteration with prominent apical snouts and secretions (CAPSS). The purpose of this study was to define the pathologic spectrum and mammographic features of these lesions. We reviewed histologic sections and mammograms from 100 consecutive breast biopsies performed for microcalcifications. The prevalence and histologic features of CAPSS and the association with other histologic findings were recorded. CAPSS was identified in 42% of cases. At the lower end of the spectrum were lesions similar to columnar alteration of lobules but in which apical cytoplasmic secretion and nuclear stratification were more pronounced and cells with a hobnail configuration were common. More advanced lesions showed columnar epithelial cell tufts, bridges, and micropapillations with prominent apical cytoplasmic snouts and with greater degrees of nuclear stratification and atypia. At the upper end of the spectrum were lesions that could arguably be considered DCIS. Calcifications were present within CAPSS in 74% of cases, were frequently psammomatous, and were typically nonbranching and often round on mammography. Columnar alteration of lobules was more common in biopsies with than without CAPSS (74 versus 36%, p < 0.001). Ductal carcinoma in situ was seen with similar frequency in biopsies with and without CAPSS (38 versus 41%). However, DCIS in cases with CAPSS was more often of the low-grade micropapillary-cribriform type than in cases without CAPSS (56 versus 17%, p < 0.01), and CAPSS and DCIS commonly coexisted in the same or adjacent TDLUs. In conclusion, 1) CAPSS encompasses a spectrum of lesions bounded at the lower end by columnar alteration of lobules and at the upper end by low grade DCIS. Lesions recently described by Page as "hypersecretory hyperplasia with atypia" fall within this spectrum. 2) Some CAPSS lesions present architectural or cytologic features that create diagnostic difficulties and raise the possibility of atypical ductal hyperplasia or DCIS; however, the level of cancer risk associated with CAPSS lesions that do not fulfill established criteria for atypical ductal hyperplasia or DCIS is unknown and requires evaluation in follow-up studies. PMID- 9850179 TI - Herpes virus type 8-negative primary effusion lymphoma associated with PAX-5 gene rearrangement and hepatitis C virus: a case report and review of the literature. AB - At present, there is no case report of HHV8- primary effusion lymphoma (PEL) with t(9;14)(p13;q32) involving both PAX-5 and immunoglobulin heavy chain gene rearrangement, which is a rare translocation in B-cell non-Hodgkin's lymphoma, in an HIV- patient. We examined an HIV-seronegative 63-year-old Japanese man with hepatitis C virus-associated liver cirrhosis and hepatocellular carcinoma manifesting peritoneal lymphomatous effusion without tumor mass at any body site. The lymphoma cells were examined twice by light microscopy, immunohistochemistry, three-color flow cytometry, cytogenetics, and molecular analyses. The nuclear morphology of lymphoma cells was similar to that of large noncleaved cells, although the lymphoma cell size was a little smaller that of the usual large-cell lymphoma. Immunophenotyping of lymphoma cells in the ascitic fluid revealed a mature peripheral B-cell phenotype (CD5- CD10- CD19+ CD20+ CD22+ Ig G+ lambda+). Cytogenetics showed a clonal population: 45,X,-Y, der(2) t(2;6)(q31;p21.3), t(4;8)(q21;q11.2), der(6) t(2;6)(q31;p21.3) add(6)(q15), t(9;14)(p13;q32.3) [10]/47, idem, +der(6) t(2;6), +16[10]. Southern blot analysis revealed rearranged fragments with a probe for immunoglobulin heavy chain, some of which were a size similar to those with a PAX-5 gene probe. Polymorphism, not rearrangement, of the c-MYC gene, was also found. HHV8 and the Epstein-Barr virus were not detected by polymerase chain reaction. This case is the first report of an HHV8- PEL with t(9;14) involving a PAX-5 gene rearrangement in an HIV seronegative patient. This primary effusion lymphoma manifested spontaneous regression without any therapy. These findings suggest that there may be an additional subcategory of primary effusion lymphoma that is not associated with HHV8 nor c-MYC(R) but is pathogenetically associated with the PAX-5 gene or hepatitis C virus. PMID- 9850180 TI - Hypothalamic hamartoma: report of a case with unusual histologic features. AB - A rare case of hypothalamic hamartoma with unusual radiologic and histopathological features is described, possibly the first of its type in English literature. A 1.5-year-old female child presented with precocious puberty. MR scan of the brain revealed a pedunculated hypothalamic mass, most of which was isointense with normal brain on T1- and T2-weighted images. However, a sizeable component of the lesion was hyperintense on T1-weighted images, suggestive of adipose tissue. Microscopically, the lesion was a hamartoma composed of an admixture of neuroectodermal elements, namely glial cells, neurons, and nerve bundles along with mesenchymal elements in the form of fibroadipose tissue. PMID- 9850181 TI - Nerve sheaths and keratin. PMID- 9850182 TI - Limitations of histologic classification of iron overload. PMID- 9850184 TI - Conversion of cardiac and liver transplant recipients from HPLC and FPIA (polyclonal) to an FPIA (monoclonal) technique for measurement of blood cyclosporin A. AB - In an effort to replace HPLC and FPIA (polyclonal) for whole blood determination of Cyclosporin A (CsA), this study examined the application of FPIA (monoclonal) in patients post cardiac and liver transplantation. The assay had a minimum detectable dose of 15 microg/L, an overall recovery of 97% and was linear to 1200 microg/L, and gave inter-assay precision values of < 5% (CV). On comparing FPIA (monoclonal) and HPLC for 59 cardiac transplant recipient blood samples, a correlation of FPIA (monoclonal) = 1.30 (HPLC) + 36.34, r = 0.96 was obtained. With liver transplant samples (n = 348), the correlation was FPIA (monoclonal) = 1.21 (HPLC) + 42.15, r = 0.98. Correlation on 131 cardiac transplant recipients gave FPIA (monoclonal) = 0.31 FPIA (polyclonal) + 43.97, r = 0.68. It is concluded that when converting from HPLC to FPIA (monoclonal) a positive bias of 21%-30% is observed, and in replacing FPIA (polyclonal) with FPIA (monoclonal), a negative bias of 50%-69% is seen with liver and cardiac patients respectively. These data indicate that therapeutic ranges should be re-established or adjustments in CsA dosing would be necessary. PMID- 9850183 TI - Immune complex transfer enzyme immunoassay for antibody IgM to HIV-1 p17 antigen. AB - The immune complex transfer enzyme immunoassay for antibody IgM to HIV-1 p17 antigen is described. Serum samples containing antibody IgM to HIV-1 p17 antigen were incubated simultaneously with 2,4-dinitrophenyl-bovine serum albumin recombinant p17 (rp17) conjugate and rp17-beta-D-galactosidase conjugate, and the immune complex formed comprising the three components was trapped onto colored polystyrene beads coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG. Subsequently, the immune complex was transferred to white polystyrene beads coated with monoclonal mouse (antihuman IgM) IgG in the presence of excess of epsilonN-2,4-dinitrophenyl-L-lysine. The signal for antibody IgM to p17 antigen was the fluorescence intensity by fluorometric assay of beta-D-galactosidase activity bound to the white polystyrene beads. The periods of time required for the formation, trapping, and transferring of the immune complex comprising the three components were more than 4 hr, 2 hr, and 3 hr, respectively. The immunoassay developed was shown to be specific by inhibition of transferring the immune complex in the presence of excess of nonspecific IgM but not IgG. Signals for antibody IgM to p17 antigen with serum samples of HIV-1 seroconversion serum panels,--that is, with serum samples in early stages of the infection--tended to be higher than those with serum samples from HIV-1 asymptomatic carriers probably long after the infection and patients with ARC and AIDS. In contrast, signals for antibody IgG to p17 antigen with serum samples of HIV-1 seroconversion serum panels tended to be higher than signals for antibody IgM to p17 antigen but were much lower than signals for antibody IgG to p17 antigen with serum samples from HIV-1 asymptomatic carriers and patients with ARC and AIDS. PMID- 9850186 TI - Analysis of urinary albumin, transferrin, N-acetyl-beta-D-glucosaminidase and beta2-microglobulin in patients with impaired glucose tolerance. AB - We investigated the changes in urinary albumin and urinary transferrin as glomerular proteins, and in urinary N-acetyl-beta-D-glucosaminidase and urinary beta2-microglobulin as tubular proteins, in patients with impaired glucose tolerance. We attempted to compare the proteins of normal subjects to those of diabetics with pre-nephropathy. Transferrin and N-acetyl-beta-D-glucosaminidase levels were significantly increased in patients with impaired glucose tolerance, while albumin and beta2-microglobulin levels were only slightly increased. In addition, there was no significant difference in transferrin levels between patients with impaired glucose tolerance and type 2 diabetics with pre nephropathy. In our observation, although albumin levels were only slightly increased in patients with impaired glucose tolerance, a sharp increase in transferrin levels was reflected in patients with glomerular disorders. In addition, since N-acetyl-beta-D-glucosaminidase levels varied markedly, tubular disorders were suspected. It should be stressed that increased parameters for both glomerular and tubular disorders in group C--patients who showed abnormal levels in three proteins--had already been observed in some patients with impaired glucose tolerance. Therefore, the evaluation of the mutual relationships between various urinary protein components in patients with impaired glucose tolerance will become a more important assessment tool than that of single urinary protein components. PMID- 9850185 TI - Sensitive enzyme immunoassay of antibodies to HIV-1 p17 antigen using indirectly immobilized recombinant p17 for diagnosis of HIV-1 infection. AB - Recombinant p17 (rp17) antigen of HIV-1 and maltose binding protein-rp17 fusion protein (MBP-rp17) were immobilized onto polystyrene beads in different ways: rp17 and MBP-rp17 were immobilized directly onto polystyrene beads by physical adsorption; biotinyl-rp17 and biotinyl-MBP-rp17 were immobilized indirectly onto streptavidin-coated polystyrene beads; and 2,4-dinitrophenyl (DNP)-MBP-rp17 was immobilized indirectly onto (anti-DNP) IgG-coated polystyrene beads. These directly and indirectly immobilized antigens were incubated with urine samples containing antibody IgG to p17 antigen and subsequently with rp17-beta-D galactosidase conjugate or (anti-human IgG gamma-chain) Fab'-beta-D-galactosidase conjugate. Beta-D-galactosidase activity bound to the polystyrene beads was assayed by fluorometry. When rp17-beta-D-galactosidase conjugate was used, signals (fluorescence intensities for bound beta-D-galactosidase activity) were much higher with the indirectly immobilized antigens than those with the directly immobilized antigens. By experiments using (anti-human IgG gamma-chain)Fab'-beta D-galactosidase conjugate, the binding of rp17-beta-D-galactosidase conjugate to antibodies against p17 antigen bound to directly immobilized rp17 antigen was shown to be seriously limited as compared with that to antibodies against p17 antigen bound to indirectly immobilized DNP-MBP-rp17. When rp17-beta-D galactosidase conjugate and serum samples were used, serum interference was much less with indirectly immobilized DNP-MBP-rp17 than with directly immobilized rp17 antigen, and the sensitivity of enzyme immunoassay for antibody IgG to p17 antigen using indirectly immobilized DNP-MBP-rp17 was 1,000- to 3,000-fold higher than that of enzyme immunoassay using directly immobilized rp17 antigen and Western blotting for p17 band. This sensitive enzyme immunoassay indicated positivity in HIV-1 seroconversion serum panels as early as conventional methods for antibodies to HIV-1 and earlier than Western blotting for p17 band. PMID- 9850187 TI - Determination of the bioavailability of intranasal elcatonin in humans: development of a sandwich transfer enzyme immunoassay for elcatonin. AB - A sandwich transfer enzyme immunoassay for elcatonin (ECT) and its usability for the pharmacokinetic study are described. The anti-salmon calcitonin (SCT) antibody was used for the present assay. The assay procedure consisted of the reaction of ECT with 2,4-dinitrophenylbiotinyl anti-SCT IgG and anti-SCT Fab' beta-D-galactosidase conjugate, trapping onto (anti-2,4-dinitrophenyl bovine serum albumin) IgG-coated polystyrene balls, eluting with epsilonN-2,4 dinitrophenyl-L-lysine and transferring to streptavidin-coated polystyrene balls and fluorometric detection of beta-D-galactosidase activity. The practical detection limit of ECT was 0.15 pg (44 amol)/50 microl of sample and 3 pg/ml as the concentration. The application of this method has enabled us to directly estimate the bioavailability of ECT dosed intranasaly at a therapeutic level (100 IU, 17 microg) for its anti-osteoporotic effect as compared to an intramuscular dose (40 IU, 6.7 microg). The pharmacokinetic parameters of the intranasal ECT (n = 6) thus estimated were as follows: the area underthe serum concentration-time curve (AUC) = 2,570 +/- 1,650 (SD) pg x min/ml, and the maximal concentration (Cmax) = 60 +/- 25 (SD) pg/ml with the maximal time (Tmax) = 17.5 +/- 6.9 (SD) min, when the AUC for the intramuscular ECT (n = 9) = 9,460 +/- 5,870 (SD) pg x min/ml and the Cmax = 165 +/- 79 (SD) pg/ml with the Tmax = 16.1 +/- 4.2 (SD) min. PMID- 9850188 TI - Reference distributions for immunoglobulins A, G, and M: a practical, simple, and clinically relevant approach in a large cohort. AB - Serum immunoglobulins are measured millions of times each year, yet clinical interpretations remain hampered by inadequate age- and gender-specific reference limits. In order to provide more reliable and comprehensive reference distributions for IgA, IgG, and IgM measurements, we analyzed automated immunoassay values from 115,017 serum samples from northern New England patients (99% Caucasian) who were tested in our laboratory between 1986 and 1995. Measurements were standardized to reference material, CRM 470 (RPPHS). A simple, practical, and clinically relevant approach was used to determine reference distributions for the immunoglobulins over a wide range of ages for males and females. Levels of IgA and IgM varied considerably by age, and by gender for IgM. For each of the analytes, the observed 5th and 95th centiles were symmetric about the median and approximately constant over the entire age range. When immunoglobulin reference values are expressed as multiples of the age- and gender specific regressed medians, the resulting distributions fit a log-Gaussian distribution well. This finding enables interpretation of serum immunoglobulin measurements using a common unit (multiples of the median) that is independent of age or gender. Insights gained from this study can help improve and simplify the interpretation of immunoglobulin measurements. PMID- 9850189 TI - Reference distributions for immunoglobulins A, G, and M: a comparison of a large cohort to the world's literature. AB - Clinical interpretation of immunoglobulin measurements requires accurate and precise assays and widely accepted reference preparations, as well as reliable age- and gender-specific reference ranges. This last requirement, the topic of numerous publications, has not been addressed adequately. By a combination of computerized and hand searching of the literature from 1961 to 1997, we identified 109 publications presenting IgA, IgG, and/or IgM reference data in healthy individuals. After eliminating studies that lacked appropriate clinical, statistical, or reference material information, data from the 17 acceptable studies were converted to a common reference material, CRM 470/RPPHS. When median levels from our recently published large cohort study are superimposed on these published medians, they fall within the ranges of reported medians. The widths of published 95 percentile reference ranges (where each individual's health was verified) were also found to agree closely with the reference range widths found in our data (where inclusion was based on the reported diagnosis). The current combined study of narrowly applied reference ranges validates our recently published age- and gender-specific reference data for immunoglobulinsA, G, and M. Those new data can now be considered as a source of reliable reference ranges to be used by laboratories when interpreting immunoglobulin measurements. PMID- 9850190 TI - Follow-up study on urinary type IV collagen in patients with early stage diabetic nephropathy. AB - Type IV collagen is a major component released from the glomerular and tubular basement membranes. To investigate the alteration of renal type IV collagen turnover in early stage diabetic nephropathy, urinary type IV collagen was measured by a highly sensitive one-step sandwich enzyme immunoassay (EIA). Urinary samples were obtained from 94 diabetic patients without overt proteinuria. Among those patients, 61 were normoalbuminuric and 33 patients were in the microalbuminuric group. Levels of urinary type IV collagen were serially examined at the start of this study and again one year later. The levels of urinary type IV collagen in patients in the microalbuminuric group were significantly higher than those in the normoalbuminuric group (P < 0.01). There was a significant correlation between the concentration of urinary albumin and urinary type IV collagen in both groups (P < 0.05). Twenty-eight patients (45.3%) in the normoalbuminuric group who showed an abnormal elevation of urinary type IV collagen in comparison to the reference range of normal healthy adults (normal range; less than 3.5 microg/g x Cr). Seven (25%) out of these 28 normoalbuminuric patients with increased urinary type IV collagen progressed to the microalbuminuric group one year later. The levels of urinary type IV collagen in such patients were significantly increased. In the 21 patients who stayed within the normoalbuminuric group, the urinary type IV collagen levels were significantly decreased one year later. It appears that the levels of urinary type IV collagen might reflect ongoing alteration of the extracellular matrix (ECM) turnover and might define more specifically the early stage diabetic nephropathy than the detection of microalbuminuria. It is concluded that the serial measurement of urinary type IV collagen can be a useful marker for detecting renal injury in diabetes. PMID- 9850191 TI - Cyanotic heart lesions with increased pulmonary blood flow. AB - This article examines common cyanotic congenital heart lesions that result in an increase in pulmonary blood flow. These lesions include transposition of the great arteries, truncus arteriosus, total anomalous pulmonary venous connection, tricuspid atresia, and single ventricle. The complications caused by increased pulmonary blood flow are examined. The hemodynamics, presentation, diagnosis, medical management, and surgical treatment of each lesion are discussed. Support and teaching needs of the parents are also addressed. PMID- 9850192 TI - CBC and differential in VLBW infants. AB - PURPOSE: This study tested the adequacy of established leukocyte indices in infants < or = 1,200 gm birth weight and < or = 32 weeks gestation. DESIGN: Charts of all admissions to the NICU from March 1991 through September 1994 were retrospectively reviewed. SAMPLE: All infants (n = 131) were outborn and admitted to the NICU within 24 hours after birth. MAIN OUTCOME VARIABLE: The leukocyte indices obtained during the first 72 hours after birth were plotted as a function of time in hours. RESULTS: Mode of delivery significantly affected total leukocyte count and absolute neutrophil count; sex and use of prenatal steroids did not. Ranges for total leukocyte count, absolute neutrophil count, and immature/total neutrophil (I:T) ratio demonstrate that normal values are variable and change over time. PRACTICE RECOMMENDATIONS: The wide distribution of values found, coupled with differences noted by mode of delivery, make normative data difficult to apply. The clinician must question the appropriateness of routinely obtaining a complete blood count and the clinical utility of the results. PMID- 9850193 TI - Nursing case management in the NICU: enhanced coordination for discharge planning. AB - A primary goal of health care today is finding alternative ways to provide high quality, cost-effective care. A model of care that addresses these issues is nursing case management. This article describes the development and implementation of nursing case management in a Level III regional center with the primary goal of enhancing coordination for discharge planning. The model involves development of clinical pathways and utilization of nurse practitioners as case managers. Decreased length of stay, charge per case, and readmission rates are demonstrated following implementation of this program. PMID- 9850194 TI - Mothers' perceptions of the care-by-parent program prior to hospital discharge of their preterm infants. AB - PURPOSE: This research was conducted to gain an understanding of the care-by parent experience for mothers of preterm infants. DESIGN: Phenomenologic methodology was used to explore the experiences of six mothers. Each participant slept overnight with her infant in a private hospital room. The mother assumed responsibility for her infant's care, knowing that nursing staff assistance was available if needed. Interviews were conducted on the following day and again after the infant had been home at least four days. SAMPLE: The participants were six mothers of preterm infants. MAIN OUTCOME VARIABLE: Mothers' experiences as they assumed responsibility in caring for their preterm infants away from the intensive care nursery. RESULTS: Findings revealed that the care-by-parent experience gives the mother an opportunity to assume full responsibility for her preterm infant's care, tests the reality of caregiving, helps her learn about caregiving activities and her infant's patterns of behavior, and confirms her readiness for independent parenting and the infant's readiness for discharge home. These findings confirm the benefits of rooming-in prior to discharge of preterm infants and provide guidelines for discharge planning. PMID- 9850195 TI - Universal hearing screening in the NICU: the Loma Linda University Children's Hospital experience. AB - The most common birth defect in the U.S. today is hearing loss, ranging from mild to profound. Even mild hearing impairment seriously affects language, speech, and cognitive development. Early identification and intervention lead to improved communication skills, which positively impact psychosocial, educational, and vocational development. Loma Linda University Children's Hospital NICU has implemented a program consistent with the Joint Committee on Infant Hearing's 1994 position statement, which advocates universal detection of infants with hearing loss, and with the National Institutes of Health 1993 position statement on early identification of hearing impairment in infants and young children, which recommends screening all infants admitted to NICUs for hearing loss prior to discharge. This article discusses the process of change required to meet these goals and outlines areas for further evaluation and change. PMID- 9850196 TI - Neonatal nurse knowledge of penicillin therapy. PMID- 9850198 TI - Visitation patterns: the parents who visit "too much". PMID- 9850199 TI - Tips to merge by. PMID- 9850197 TI - Motherhood by contract? PMID- 9850200 TI - The National Health Service celebrates its 50th birthday. PMID- 9850201 TI - British psychiatric morbidity survey. PMID- 9850202 TI - Severe personality disorder--whose responsibility? PMID- 9850203 TI - Excess mortality of mental disorder. AB - BACKGROUND: We describe the increased risk of premature death from natural and from unnatural causes for the common mental disorders. METHOD: With a Medline search (1966-1995) we found 152 English language reports on the mortality of mental disorder which met our inclusion criteria. From these reports, covering 27 mental disorder categories and eight treatment categories, we calculated standardised mortality ratios (SMRs) and 95% confidence intervals (CIs) for all causes of death, all natural causes and all unnatural causes; and for most, SMRs for suicide, other violent causes and specific natural causes. RESULTS: Highest risks of premature death, from both natural and unnatural causes, are for substance abuse and eating disorders. Risk of death from unnatural causes is especially high for the functional disorders, particularly schizophrenia and major depression. Deaths from natural causes are markedly increased for organic mental disorders, mental retardation and epilepsy. CONCLUSION: All mental disorders have an increased risk of premature death. PMID- 9850204 TI - Sertraline in the treatment of panic disorder. A multi-site, double-blind, placebo-controlled, fixed-dose investigation. AB - BACKGROUND: This study compared the efficacy and safety of sertraline to placebo in treating panic disorder. METHOD: 178 out-patients with panic disorder who exhibited at least four panic attacks during the four weeks prior to screening and three during the two weeks of lead-in were randomly assigned to 12 weeks of double-blind treatment with sertraline (50, 100 or 200 mg) or placebo. RESULTS: Sertraline was superior to placebo in reducing the number of panic attacks, situational attacks, unexpected attacks, limited symptom attacks, and time spent worrying (all P < 0.01) and the Hamilton Anxiety Scale (P < 0.05), although Clinical Global Impression (Improvement) did not significantly differentiate groups at 12 weeks and at end-point. No serious adverse events were associated with sertraline. No dose relationship was found for adverse events; overall drop out rates were not different for sertraline or placebo, although more sertraline treated subjects discontinued for adverse events, typically early in the study. Only dry mouth and ejaculation failure (primarily ejaculation delay) were associated significantly with sertraline. CONCLUSIONS: Sertraline was effective and safe in reducing panic attacks. Higher doses were no more effective than the 50 mg dose. PMID- 9850205 TI - London-east Anglia randomised controlled trial of cognitive-behavioural therapy for psychosis. III: Follow-up and economic evaluation at 18 months. AB - BACKGROUND: A randomised controlled trial of cognitive-behavioural therapy (CBT) for people with medication-resistant psychosis showed improvements in overall symptomatology after nine months of treatment; good outcome was strongly predicted by a measure of cognitive flexibility concerning delusions. The present paper presents a follow-up evaluation 18 months after baseline. METHOD: Forty seven (78% of original n = 60) participants were available for follow-up at 18 months, and were reassessed on all the original outcome measures (see Part I). An economic evaluation was also completed. RESULTS: Those in the CBT treatment group showed a significant and continuing improvement in Brief Psychiatric Rating Scale scores, whereas the control group did not change from baseline. Delusional distress and the frequency of hallucinations were also significantly reduced in the CBT group. The costs of CBT appear to have been offset by reductions in service utilisation and associated costs during follow-up. CONCLUSIONS: Improvement in overall symptoms was maintained in the CBT group 18 months after baseline and nine months after intensive therapy was completed. CBT may be a specific and cost-effective intervention in medication-resistant psychosis. PMID- 9850206 TI - SPET study of verbal fluency in schizophrenia and epilepsy. AB - BACKGROUND: The association between temporal lobe epilepsy and schizophrenia suggests that the critical abnormality may be pathology within the temporal lobes. People with schizophrenia-like psychosis of epilepsy (SLPE) provide a useful group in which to examine the importance of temporal and frontal lobe dysfunction in schizophrenia. METHOD: A verbal fluency activation paradigm and a 99mTc HMPAO SPET were used to study frontotemporal function in people with SLPE (n = 12), schizophrenia (n = 11) and epilepsy (n = 16). RESULTS: People with SLPE differed from both other groups by showing lower blood flow in the left superior temporal gyrus during performance of a verbal fluency task compared with a word repetition task (F = 5.4, P = 0.01). During the verbal fluency task people with primary schizophrenia showed a greater increase in blood flow in anterior cingulate (F = 4.5, P = 0.02) than the other two groups. There were no between group differences in frontal brain regions. CONCLUSION: Our findings support an association between left temporal lobe abnormality and SLPE. The different patterns of activation observed in people with primary schizophrenia and SLPE suggests that different pathophysiological mechanisms may operate in these two groups. In SLPE the pathophysiology may be relatively confined to the dominant temporal lobe. PMID- 9850207 TI - Genetic epidemiology of binging and vomiting. AB - BACKGROUND: Bulimia nervosa is typically defined as the combination of the behaviours of binging and vomiting. We sought to clarify the relationship of these behaviours from a genetic epidemiological perspective. METHOD: Using data on the lifetime history of binging and vomiting from a personally interviewed population-based sample of female twins (n = 1897), we applied bivariate twin modelling to estimate the sources of variation for these traits. RESULTS: The association between having ever binged (23.6%) and having ever induced vomiting (4.8%) was very strong (odds ratio = 8.78, P << 0.0001). The best-fitting model indicated that lifetime binging and vomiting were both heritable (46% and 72%) and influenced by individual-specific environmental factors (54% and 28%). The overlap between the genetic (ra = 0.74) and individual-specific environmental factors (re = 0.48) for the two traits was substantial. No violations of the equal environmental assumption were evident. CONCLUSIONS: Including binging and vomiting under the rubric of bulimia nervosa appears to be appropriate. Our data are consistent with the identification of binging and vomiting as complex traits resulting from the interplay of multiple genes and individual-specific environmental influences. In contrast to 'environmentalist' theories, our results suggest that genetic influences may be of particular relevance to the aetiology of binging and vomiting. PMID- 9850208 TI - Predictors of a healthy workplace for Swedish and English psychiatrists. AB - BACKGROUND: Few studies have attempted to analyse the organisational aspects of the workplace that may affect the well-being of psychiatrists and even fewer have offered insights into what a positive workplace might look like. This study provides an outline of such a workplace, with reference to individual and organisational factors. In addition, a comparison is made between two European cities to check for cultural differences. METHOD: Three hundred and eighty psychiatrists from Stockholm and Birmingham responded to a previously tested questionnaire on their work environment. RESULTS: Predictors of a positive workplace for psychiatrists were: high self-esteem, satisfactory support with work-related problems, lower perceived workload, positive view of leadership, low work-related exhaustion and having a sense of participation in the organisation. There were few cultural differences. CONCLUSIONS: Measures should be taken to improve leadership skills for managers, to offer more support for work-related problems, and to allow psychiatrists to participate more in the organisation. PMID- 9850209 TI - Prescribing of atypical antipsychotics. PMID- 9850210 TI - Prescribing of atypical antipsychotics. PMID- 9850211 TI - Irish ethnicity and mental health. PMID- 9850212 TI - Irish ethnicity and mental health. PMID- 9850213 TI - Risk of sudden death on high-dose antipsychotic medication: QTc dispersion. PMID- 9850214 TI - Ethnicity and clozapine metabolism. PMID- 9850215 TI - Comprehensiveness of systematic review. PMID- 9850216 TI - Qualitative research--a rejoinder. PMID- 9850217 TI - Active placebos in antidepressant trials. PMID- 9850218 TI - Establishing the limits of normal cerebral ageing and senile dementias. PMID- 9850219 TI - Loss of consciousness and post-traumatic stress disorder. A clue to aetiology and treatment. PMID- 9850220 TI - Inequalities in mental health. PMID- 9850221 TI - Peer review and editorial decision-making. AB - INTRODUCTION: This paper describes and analyses the editor's decision-making process at the British Journal of Psychiatry (BJP), and investigates the association between reviewers' assessments and editorial decisions. METHOD: Four hundred consecutive manuscripts submitted over a six-month period to the BJP were examined prospectively for assessors' comments and editorial decisions on acceptance or rejection. Interrater reliability of assessments was calculated and a logistic regression analysis investigated the effect of the rank allocated by assessors and the comprehensiveness of the assessments on the editor's decision. RESULTS: The editor sent 248/400 (62%) manuscripts to assessors for peer review. Kappa for reliability of assessors rankings was 0.1 indicating poor interrater reliability. Assessors agreed best on whether to reject a paper. A ranking of five (indicating rejection) had the greatest association with editor's rejection (P < 0.001, odds ratio 0.079), and the mean ranking of assessments was also significantly associated with editorial acceptance or rejection (P = 0.004, odds ratio 0.24). CONCLUSION: Assessors and editors tend to agree on what is clearly not acceptable for publication but there is less agreement on what is suitable. PMID- 9850224 TI - Burden of disease. Methods of calculating disability from mental disorder. AB - BACKGROUND: The Global Burden of Disease studies are important because they encompass morbidity as well as mortality. Burden due to morbidity is calculated from incidence, duration and disability. There is a dearth of epidemiological measurements of disability. METHOD: Data from a quasi-community sample (n = 1364) were analysed. Diagnoses of mental and physical disorders, and reports of disability, were based on established methods. RESULTS: The disabilities reported in mental and physical disorders were comparable. Disability was correlated with comorbidity. The disability in mental disorders was examined by three methods: pure disorders, main problem and regression. It appears that major depression and substance disorder weights were overestimated, and anxiety disorder weights were underestimated in the Global Burden of Disease studies. CONCLUSIONS: A method for disentangling the effects of concurrent comorbidity is presented. The size of burden attributed to mental disorders is of potential benefit for funding mental health services, It is important that we get the estimates right. PMID- 9850223 TI - Reduced levels of GABA-benzodiazepine receptor in alcohol dependency in the absence of grey matter atrophy. AB - BACKGROUND: We tested the hypothesis that reduced levels of the GABA benzodiazepine receptor occur in alcohol dependency using single photon emission tomography (SPET) and the specific GABA-benzodiazepine ligand, 123I-iomazenil. METHOD: Neurologically and cognitively unimpaired abstinent alcohol-dependent (n = 12) and non-alcohol-dependent male subject (n = 14) underwent a 123I-iomazenil SPET scan. SPET and magnetic resonance images were co-registered and voxel-based statistical tests performed. Subjects' clinical and alcohol history were obtained with standard questionnaires. The relationships between clinical and alcohol variables and the regional level of GABA-benzodiazepine receptors were investigated using multiple regression analysis. RESULTS: Abstinent alcohol dependent subjects had decreased levels of GABA-benzodiazepine receptor compared with non-alcohol-dependent subjects within the frontal, parietal and temporal cortices, including regions in which grey matter atrophy was absent. CONCLUSIONS: Alcohol dependency is associated with reduced GABA-benzodiazepine receptor levels in the absence of grey matter atrophy in some cortical regions, such as within the parietal lobe. Regional variability of reduction in GABA-benzodiazepine receptors demonstrates that alcohol does not have a global, toxic effect on the brain. PMID- 9850225 TI - Brain changes in schizophrenia. Volumetric MRI study of families multiply affected with schizophrenia--the Maudsley Family Study 5. AB - BACKGROUND: Structural brain abnormalities have been reported in schizophrenia. We tested the hypothesis that these abnormalities represented a marker for the genetic liability to schizophrenia in a sample of people with schizophrenia and their relatives from families multiply affected with the disorder. METHOD: We compared 31 people with schizophrenia, 57 relatives and 39 unrelated control subjects. Volumetric measurement of brain structures was carried out using stereological principles from three-dimensional reconstructed magnetic resonance images. RESULTS: Subjects with schizophrenia had larger lateral ventricles than their relatives and the normal control subjects. Relatives who were 'presumed obligate carriers' had larger left lateral ventricles than other relatives and the control subjects. Subjects with schizophrenia showed smaller whole brain and cerebellar volumes and larger lateral ventricles than their age- and gender matched unaffected siblings. CONCLUSIONS: In families multiply affected with schizophrenia lateral ventricular enlargement distinguishes people with schizophrenia and presumed obligate carriers from other relatives and unrelated control subjects. These changes may be a marker for a genetic liability to schizophrenia. PMID- 9850226 TI - People with schizophrenia and their families. Fifteen-year outcome. AB - BACKGROUND: This paper presents 15-year outcome data on place of residence, clinical and social morbidity and carer distress in a cohort of 179 people with schizophrenia who were living with their families in 1981-1982. METHOD: Data on morbidity and carer distress were derived from replicated patient and informant interviews using standardised research instruments. Details of residence came from patient and informant interviews corroborated by case note information. RESULTS: Thirty-nine (22%) of the original 179 patients were dead and six (3%) lost to follow-up. Of the remaining 134 patients, 74 (55%) still lived with their families, 31 (23%) lived in institutional accommodation, 26 (19%) lived alone and 3 (2%) were homeless. There was little change in the level of clinical or social morbidity or carer distress. CONCLUSIONS: These findings suggest that most people with schizophrenia who live with their families remain significantly disabled by their illness, while their carers suffer ongoing distress. Mental health services should seek to support families who want to care for relatives with schizophrenia. Appropriate alternative accommodation must be provided when family care is not possible. PMID- 9850227 TI - 'Pfropfschizophrenie' revisited. Schizophrenia in people with mild learning disability. AB - BACKGROUND: It is reported that people with mild learning disability have a higher point prevalence of schizophrenia than the normal population, the reasons for which are unclear. METHOD: Thirty-nine subjects with mild learning disability and schizophrenia, 34 control subjects with schizophrenia and 28 control subjects with mild learning disability were seen. Interviews with relatives and carers were also conducted. Assessments were made of clinical variables, psychopathology, neurological 'soft' signs, IQ, memory and family history. Blood was taken for karyotypic analysis from comorbid subjects. RESULTS: The comorbid group had more negative symptoms, episodic memory deficits, soft neurological signs, epilepsy and receive more community supports than control subjects with schizophrenia. Comorbid subjects had a tendency to belong to multiply affected families and show high rates of chromosomal variants on routine karyotypic testing. CONCLUSIONS: Future work on the generality of schizophrenia should include people with premorbid learning disability, as a discrete subtype from whom valuable genetic aetiological clues may be obtained. PMID- 9850228 TI - Suicidal behaviours in vulnerable adolescents. Time trends and their correlates. AB - BACKGROUND: Suicide rates have increased over the past three decades, especially in young men. Depression, conduct disorder, crime and substance misuse have also increased. This study tested hypotheses on the possible links between the secular increase in the rates of these behaviours. METHOD: A data set on 6091 subjects aged 8-18 years (58.4% boys) referred to psychiatric services over a 21-year period (1970-1990) was used. A detailed analysis of a random sample of 80 case notes was conducted. RESULTS: Suicidal behaviours increased significantly among pubertal male adolescents only (n = 1313). In this sub-sample, substance misuse accounted for the increase over time. The rates of both suicidal behaviours and of substance misuse almost doubled between 1979 and 1990 in this patient group. The case note analysis showed that solvent and alcohol misuse had also increased over the study period. Moreover, among the subjects misusing substances, alcohol was the only substance with a strong and positive association with suicidal behaviours. Substance misuse pre-dated suicidal behaviours in most patients. CONCLUSIONS: A link has been found between the increase over time of suicidal behaviors in adolescent boys and a contemporaneous increase in substance misuse. The strength and direction of the association suggests that alcohol misuse is the causal factor. PMID- 9850229 TI - Economic burden of drug dependency. Social costs incurred by drug users at intake to the National Treatment Outcome Research Study. AB - BACKGROUND: The use of illegal drugs is seen as a major social problem. The social costs can be high. METHODS: Self-report data from interviews at intake to the National Treatment Outcome Research Study (NTORS) for 1075 drug users and cost data from various sources were used to estimate criminal behaviour and health and addiction service costs for a 12-month period. Multivariate statistical analysis was used to analyse cost variations. RESULTS: Total costs for one year for the drug users amounted to over 12 million Pounds, the majority attributable to self-reported criminal behaviour. Social costs were positively related to a variety of factors including instability in living circumstances, amount of heroin used and whether or not drugs were taken intravenously. CONCLUSIONS: The study clearly demonstrates the economic and social burden associated with heavy drug users and highlights the need for further investigations into the costs and benefits of policies that can reduce these social costs. PMID- 9850230 TI - Substance use, health and social problems of service users at 54 drug treatment agencies. Intake data from the National Treatment Outcome Research Study. AB - BACKGROUND: The National Treatment Outcome Research Study (NTORS) is the first large-scale, prospective, multi-site treatment outcome study of drug users in the UK. METHOD: Substance use, health and social problems of 1075 service users were assessed at intake to 54 agencies chosen to be representative of the main national drug treatment modalities. RESULTS: Heroin dependence was the most frequently reported problem often with poly-drug and alcohol problems. Most service users injected drugs and a quarter of the injectors shared injecting equipment. Poly-drug use and heavy drinking were more common among service users admitted to residential treatments. High criminality rates were reported. Psychological and physical health problems were common and many service users had prior contact with psychiatric and medical services. CONCLUSIONS: The range and severity of problems adversely affect individual users, their families, and present a challenge to addiction treatment services. These problems create costs for the health care, social service and criminal justice system responses. PMID- 9850231 TI - Objectivity in psychoanalytic judgements. AB - BACKGROUND: There is widespread scepticism concerning the reliability and validity psychoanalytic judgements of patient-therapist transactions. We predicted that (a) in reviewing the initial part of 14 videotaped assessment interviews with borderline and dysthymic subjects, dynamic psychotherapists would agree in their ratings of psychoanalytically relevant characteristics of subjects' interpersonal relations; (b) inter-correlations among the ratings would conform with those expected from psychoanalytic descriptions of paranoid schizoid' and 'depressive position' states of mind; and (c) these ratings would differentiate between borderline and dysthymic groups. METHOD: Six trained psychotherapists who were blind to the design of the study, independently rated qualities of interpersonal relatedness during the first 30 minutes of each interview, on a 30-item 'personal relatedness profile'. RESULTS: There was satisfactory interrater reliability in judgements among the raters, and evidence that the items were interrelated. There was also a significant difference between the two subject groups. CONCLUSIONS: It is possible to make reliable psychoanalytic judgements about qualities of interpersonal relatedness. Moreover, there is evidence that paranoid-schizoid and depressive positive aspects of psychological functioning do constitute a meaningful constellation of clinically grounded phenomena. PMID- 9850232 TI - Headings in structured abstracts. PMID- 9850233 TI - Recovered memories of childhood sexual abuse. PMID- 9850234 TI - Recovered memories of childhood sexual abuse. PMID- 9850235 TI - Recovered memories of childhood sexual abuse. PMID- 9850236 TI - Antidepressant quandaries. PMID- 9850237 TI - Antidepressant quandaries. PMID- 9850238 TI - Antidepressant quandaries. PMID- 9850239 TI - Aggression and violence in severe mental illness. PMID- 9850240 TI - Prenatal exposure to maternal stress and subsequent schizophrenia. PMID- 9850241 TI - Evolution of schizotypy. PMID- 9850242 TI - Semantic priming in schizophrenia. PMID- 9850243 TI - Are first-rank symptoms encryption errors. PMID- 9850244 TI - Lifetime risk of suicide in affective disorders. PMID- 9850245 TI - Suicide, country of birth and coroner's verdicts. PMID- 9850246 TI - Opportunities for psychiatry from genetic findings--some concerns. PMID- 9850247 TI - Fluoxetine-terfenadine and sexual dysfunction. PMID- 9850248 TI - Graft rejection. PMID- 9850249 TI - Ophthalmic pathology in the electronic age. PMID- 9850250 TI - The use of combined intravenous pulse methylprednisolone and oral cyclosporin A in the treatment of corneal graft rejection: a preliminary study. AB - PURPOSE: Oral cyclosporin A used in addition to high-dose intravenous pulse methylprednisolone has been shown to have an adjunctive effect in reversing the rejection of liver and renal transplants. The aim of this prospective study was to evaluate the benefits and risks of this combined drug therapy in acute corneal graft rejection. METHODS: Eleven patients with acute corneal graft rejection received the combined regimen of a single pulse of intravenous methylprednisolone (500 mg) and a low dose of oral cyclosporin A (to maintain a trough blood level of 100-200 micrograms/l). RESULTS: At a mean follow-up of 16.5 months (range 8-22 months) from the presentation of the graft rejection, reversal of graft rejection was achieved in 10 of 11 cases (90.9%). No recurrence of graft rejection was encountered during the study period. One patient developed a duodenal ulcer, which healed after medical treatment. No other complications were encountered. CONCLUSIONS: The high efficacy and low risk of the combined regimen demonstrated in this preliminary study call for a larger-scale prospective double-masked study to confirm the value of this treatment protocol. PMID- 9850251 TI - Corneal endothelial specular microscopy following deep lamellar keratoplasty with lyophilised tissue. AB - PURPOSE: To assess the corneal endothelial cell density following deep lamellar keratoplasty (DLK) carried out by air dissection and with the transplantation of lyophilised tissue. METHOD: Contact endothelial specular microscopy was carried out on a series of patients who had undergone DLK with a minimum of 1 year post operative follow-up. RESULTS: Twenty eyes of 18 patients were examined at follow up times ranging from 1 to 8 years post-operatively (mean 3 years). Indications for surgery were: keratoconus (n = 12), herpes simplex virus (HSV) keratitis (n = 1), herpes zoster ophthalmicus (HZO) keratitis (n = 1), lipid keratopathy (n = 2), lattice dystrophy (n = 1) and corneal scarring (n = 3). Overall, mean post operative cell density was 2417 cells/mm2 (range 928-3656 cells/mm2). In eyes with pathological conditions not likely to have affected the endothelial cell density, such as keratoconus or lattice degeneration, the mean cell density was 2837 cells/mm2 (range 1030-3656 cells/mm2). CONCLUSIONS: In patients undergoing DLK for conditions such as keratoconus the post-operative cell density was at a normal level in the majority of cases. Cell loss through this surgical intervention thus appears generally to be small. The prospect for long-term survival of these grafts is good. PMID- 9850252 TI - Surgical management of upper lid epiblepharon. AB - BACKGROUND: Upper lid epiblepharon refers to a developmental anomaly characterised by the presence of an exaggerated skin fold running horizontally across the upper lid below the lid margin, which in severe cases is associated with lash-cornea touch or even corneal punctate epithelial erosions. METHOD: This retrospective study looked at 14 patients who underwent bilateral upper lid pretarsal orbicularis debulking and skin excision to correct lash-globe contact from the upper lid. RESULTS: Surgery was successful in 25 of 28 lids, with one recurrence being successfully re-operated. The remaining 2 recurrences did not undergo further surgery as the patients were asymptomatic. CONCLUSION: This surgical procedure appears to be an effective treatment of lash inversion in upper lid epiblepharon with good aesthetic results. PMID- 9850253 TI - Lymphangiectasia haemorrhagica conjunctivae. AB - PURPOSE: To re-describe a condition that has not been mentioned in the literature for more than four decades and to outline a new method of treatment of the pathology using an argon laser. METHODS: Nine patients were identified over a 2 year period who fitted a clinical picture consistent with the features of lymphangiectasia haemorrhagica. Three were treated for frequent recurrent haemorrhages with a newly developed technique using the argon laser and followed for up to 2 years. RESULTS: None of the three treated patients has had a recurrence of the condition to date. In the six untreated patients the condition resolved spontaneously after repeated attacks over between 8 months and 8 years. CONCLUSIONS: Lymphangiectasia haemorrhagica conjunctivae is probably more common than expected and may resolve spontaneously, but is readily amenable to treatment with the argon laser if attacks become frequent, uncomfortable or unsightly. PMID- 9850255 TI - First day post-operative review following uncomplicated phacoemulsification. AB - PURPOSE: To determine whether the first day review is essential in management of patients following uncomplicated phacoemulsification with intraocular lens implantation. Patients are routinely seen on the day following surgery. This can produce logistical problems in patient attendance that can necessitate an overnight stay. If the first day review were abandoned this would lead to an increased uptake of day case surgery and a reduction in health care costs. METHODS: A retrospective cohort study was performed on all cases of uncomplicated phacoemulsification with intraocular lens implant surgery over a 6 month period. Slit lamp examination findings on the first post-operative day were reviewed. Visual acuity, corneal clarity, anterior chamber activity, intraocular pressure and configuration of the pupil were recorded. Any cases that failed to meet predetermined criteria underwent full case-note review. The subsequent management of these patients was analysed. RESULTS: Of the 201 cases of uncomplicated surgery, 74 cases (37%) failed to meet the study criteria and underwent review. Of these only 12 (6%) had their management altered as a result of the first day post-operative findings. This was entirely due to raised intraocular pressure. Visual acuity, corneal oedema and activity in the anterior chamber all improved on subsequent follow-up. CONCLUSIONS: The first day review of uncomplicated phacoemulsification with lens implant surgery provides the opportunity to treat raised intraocular pressure. A prospective randomised study is needed to identify means to prevent the post-operative intraocular pressure rise before we are able to consider abandoning the first review. PMID- 9850254 TI - Early induced astigmatism following phacoemulsification and flexible lens implantation through an oblique corneal tunnel. AB - PURPOSE: To analyse the early surgically induced corneal astigmatism (SICA) with a 3.5 mm oblique clear corneal incision (CCI) for phacoemulsification and flexible silicone implant. METHODS: Sixty-four consecutive patients were included in this study. The keratometry readings were recorded on the day prior to surgery, the first day post-operatively and at 6 weeks. The SICA was evaluated according to four different methods: (i) simple subtraction, (ii) vector analysis, (iii) vector decomposition and (iv) Naeser polar values. RESULTS: At 6 weeks post-operatively the SICA was equally divided between with-the-rule (WTR) and against-the-rule (ATR) astigmatism, according to the vector analysis in combination with vector decomposition. Both the simple subtraction method and Naeser polar values were misleading. CONCLUSION: The oblique clear corneal section for phacoemulsification is a comfortable and safe incision with a predictable and acceptable degree of SICA that may be better tolerated than that induced by superior incision. PMID- 9850256 TI - Is eye padding routinely necessary after uncomplicated phacoemulsification? AB - PURPOSE: To investigate the value of eye padding following uncomplicated phacoemulsification under peribulbar anaesthesia. METHODS: A prospective randomised controlled study was conducted to compare the effect of a conventional eye pad and shield with that of a clear eye shield applied without a pad in 83 patients undergoing routine phacoemulsification under peribulbar anaesthesia without lid block. The primary outcome measures were corneal fluorescein staining, discomfort, diplopia and mobility. RESULTS: Moderate or severe corneal fluorescein staining on the first post-operative day was significantly more common in the pad and shield group (39%) than in the clear shield group (19%) (p < 0.01). There was no significant difference in post-operative pain as measured either by visual analogue scale or by categorical pain scale. Forty per cent of the clear shield group reported transient post-operative diplopia during the immediate post-operative period compared with 7% of the pad and shield group (p < 0.001). There was no significant difference in reported mobility between the two groups. CONCLUSIONS: Following phacoemulsification under peribulbar anaesthesia, the use of a gauze eye pad is associated with greater corneal fluorescein staining than a clear plastic shield without pad and offers no reduction in discomfort. A clear shield protects the globe against direct trauma, is associated with reduced moderate to severe corneal staining and facilitates vision in the early post-operative period. Transient diplopia reported by some patients given a clear shield is not disabling and would not be expected to occur in patients with one seeing eye. The use of a clear shield alone is a safe alternative to eye padding and offers important advantages in patients with one seeing eye. PMID- 9850257 TI - The management of dislocated lens material after phacoemulsification. AB - PURPOSE: To determine the visual outcome, prognostic factors, and effect of timing of surgical intervention in patients with retained lens fragments after phacoemulsification. METHODS: A retrospective review was carried out of the notes of 44 consecutive patients who suffered posterior dislocation of lens fragments during phacoemulsification, of whom 34 underwent vitrectomy. RESULTS: The presence of severe uveitis at presentation was a significant predictor of a final visual acuity of less than 6/60 (p = 0.002). A raised intraocular pressure at presentation (> 29 mmHg) significantly increased the risk of chronic glaucoma (p = 0.0279), but there was no association between glaucoma and delay in vitrectomy. Patients operated on within 1 week of cataract surgery obtained a better visual outcome (64.7%) of patients achieved 6/12 or better visual acuity) than those operated on later than 1 week (41% obtained 6/12), but this difference did not reach statistical significance. CONCLUSIONS: The trend towards a better visual outcome with early vitrectomy was not statistically significant. A large prospective trial is indicated to determine the optimum time for vitrectomy in these patients. PMID- 9850258 TI - Indocyanine green angiographic features of traumatic choroidal ruptures. AB - PURPOSE: To investigate the indocyanine green (ICG) angiographic findings of healed traumatic choroidal ruptures. METHODS: Three eyes of two patients with choroidal ruptures underwent both fluorescein angiography and ICG angiography with a digital high-resolution system. RESULTS: Fundus examination showed healed choroidal ruptures in all three eyes. Two of the eyes had old macular scars and one eye had a recent choroidal neovascular membrane with serous detachment of the macula. On fluorescein angiography, the choroidal ruptures showed hypofluorescence at very early stages followed by late hyperfluorescence. On ICG angiography, the choroidal ruptures were hypofluorescent in early, mid and late phases. ICG angiography provided a clear delineation of the choroidal ruptures. In all eyes, the lesion was better defined on the ICG angiogram than on the fluorescein angiogram. CONCLUSIONS: ICG angiography can be helpful in the clinical evaluation of choroidal ruptures and can be used together with fluorescein angiography for the diagnosis and follow-up of choroidal ruptures. PMID- 9850259 TI - Visual function and patient satisfaction after macular hole surgery. AB - PURPOSE: Over recent years success in macular hole surgery has increased in terms of anatomical closure. However, debate still continues on the benefit to the patient in terms of visual outcome. We designed a prospective study to investigate the outcome of full thickness macular hole (FTMH) surgery in terms of anatomical closure, visual outcome, incidence of complications and patient satisfaction. METHODS: Thirty eyes of 30 consecutive patients with FTMH were prospectively studied (stage 2 = 2; stage 3 = 23; stage 4 = 5). All cases had surgery involving vitrectomy, injection of an autologous platelet aggregate over the hole and gas tamponade. At 3 months post-operatively all cases were assessed for closure of the FTMH, Snellen acuity and the incidence of complications. At this stage all patients completed a patient satisfaction questionnaire. RESULTS: Anatomical closure of the hole was achieved in 83% of cases (25/30). Visual improvement of 2 Snellen lines or more occurred in 50% of cases (15/30). A vision of 6/12 or better was achieved in 27% of cases (8/30). A post-operative lens opacity was present in 46% (13/28) of phakic patients and a temporal, peripheral wedge-shaped field defect occurred in 17% (5/30) of cases. In this study, 53%, 70%, 57% and 67% of patients gave a positive response to specific questions about satisfaction with near, intermediate and distance vision and overall visual function respectively. CONCLUSIONS: Although the anatomical success of FTMH surgery is high the functional outcome in terms of Snellen acuity is less rewarding. Analysis of patient satisfaction suggests that the arbitrary visual outcome measures presently used may underestimate the functional benefit to the patient. Improved objective measures of visual outcome are required to assess the benefit of surgery in these cases. PMID- 9850260 TI - Microemboli are not a prerequisite in retinal artery occlusive diseases. AB - PURPOSE: Retinal artery occlusion (RAO) is caused by arterio-arterial or cardiovascular emboli in about 50% of all cases, but the role of non-embolic causes remains unclear. SUBJECTS AND METHODS: We studied 27 patients with amaurosis fugax (AFX), branch retinal artery occlusion (BRAO), central retinal artery occlusion (CRAO) and anterior ischaemic optic neuropathy (AION). Patients underwent an evaluation of cerebrovascular and cardiovascular risk factors, measurement of haemorheological parameters, and Doppler/duplex sonography including ultrasound detection of cerebral microembolic signals and echocardiography. RESULTS: Forty-one per cent of the patients had internal carotid atherosclerosis but only one patient had microembolic signals, probably due to a cardiac thrombus. Vascular risk factors, especially hypertension, were present in 82% of the patients correlating with abnormal haemorheological parameters such as increased thrombocyte reactivity. CONCLUSIONS: Our results indicate that altered haemorheological parameters, especially increased thrombocyte reactivity and vascular risk factors such as arterial hypertension, are non-embolic causes of vascular disease in a significant number of patients with RAO. This should guide diagnostic and therapeutic considerations concerning RAO in cases without proven embolic sources. PMID- 9850261 TI - Referral patterns, treatment management and visual outcome in keratoconus. AB - PURPOSE/METHOD: To determine the patterns of referral, the hospital-based management and the subjective degree of visual success achieved by keratoconics attending the Corneal Service at Sunderland Eye Infirmary, England, a specifically designed anonymous questionnaire was mailed to 160 keratoconus subjects. RESULTS: The following responses were obtained from 97 (61%) respondents: mean age of first referral (22.88 +/- 8.36 years); gender (male:female ratio 3:1); source of initial referral (optometrists 90%); visual symptoms experienced before referral (46% complained of blurred/distorted vision); and the type of visual correction before referral (70% wore a refractive correction: 72% spectacles, 28% contact lenses). Initial post-referral treatment included bilateral contact lens fitting (65%), monocular contact lens correction (26%), penetrating keratoplasty (8%) and no intervention (1%). Two-thirds of subjects were presently wearing contact lenses (67%), with the mean period of successful lens wear being 8.26 +/- 7.92 years, and 31% had undergone penetrating keratoplasty (PK) since initial referral. A subjective evaluation of visual acuity with contact lenses was made by respondents using a visual analogue scale. The mean value was 86.86 +/- 15.25 (range 50-100 mm), with 50 mm representing moderate visual acuity. Of those who underwent PK the mean subjective evaluation of the change in post-operative vision compared with pre-operatively was 82.41 +/ 26.57 (range 1-100 mm), with 50 mm representing no visual change. The majority of subjects therefore noted a moderate to marked visual improvement following PK. CONCLUSION: In this fairly young group of patients, predominantly male, the majority wore a refractive correction for approximately 10 years before referral to the Hospital Eye Service (HES). Most referrals were initiated by optometrists. On initial HES assessment the treatment for two-thirds was bilateral contact lens fitting; only a quarter were fitted monocularly. However, by the time of this study only a tenth retained monocular contact lens correction, with the mean length of time from diagnosis to bilateral contact lens wearing being approximately 5 1/2 years. The progressive nature of this disease is further evidenced by the large number of contact lens sets required by patients (approximately 5) and by the number of patients (n = 22) requiring PK, the mean length of time from diagnosis to corneal graft being approximately 8 1/2 years. PMID- 9850262 TI - The natural history of epiphora in childhood. AB - PURPOSE: To investigate the incidence and aetiology of epiphora in a population of 7-year-old children and determine whether a conservative management policy for congenital nasolacrimal duct obstruction (CNLDO) results in excess lacrimal dysfunction in later childhood. METHODS: A postal questionnaire was sent to the parents of a large cohort of 7-year-old children previously investigated to determine the incidence and natural progression of CNLDO. They were asked about the presence of epiphora, and its relationship to atopic disease and the presence of upper respiratory tract infections (URTI). Information on epiphora had also been gathered from the same cohort at age 3 1/2 years at a routine examination. Data were compared with those for epiphora in infancy in the same cohort. RESULTS: The incidence of epiphora at 3 1/2 years was 5.5%, and 7 years 7.7%. At age 7 years 70% of cases were related to atopic disease or URTI. CNLDO was not significantly related to epiphora in later childhood (p = 0.000032). CONCLUSIONS: A policy of delaying nasolacrimal probing in CNLDO until after the age of 1 year does not result in a detectable excess of lacrimal dysfunction in later childhood, when epiphora is more likely to be related to atopic disease or upper respiratory tract infection. PMID- 9850263 TI - ENT pathology and diplopia. AB - ENT pathology causes diplopia in a minority of patients. Because the various ENT conditions that can produce diplopia are relatively unusual they often escape early detection. We describe the various ENT disorders that can produce diplopia and illustrate this with our experience between 1992 and 1998. We wish to emphasise the benefit which can be obtained from undertaking an ENT history and examination in arriving at the correct diagnosis. PMID- 9850264 TI - Posterior sub-Tenon's triamcinolone injections in the treatment of uveitis. AB - PURPOSE: This is the first study reported in the United Kingdom to investigate the efficacy and safety of posterior, sub-Tenon's triamcinolone acetonide injections in the treatment of posterior and intermediate uveitis. METHODS: Twenty-eight posterior sub-Tenon's triamcinolone injections (40 mg) were given and the results analysed with a 6 month prospective follow-up in 13 cases. RESULTS: At 6 weeks follow-up, objective improvement in visual acuity occurred in 25 eyes (p < 0.05). Vitreous cellular activity was diminished in 21 eyes (p < 0.05). In most cases improvement was observed within 2 weeks of injection. No patient required repeat triamcinolone injection within 3 months and all patients previously treated with systemic immunosuppression were able to decrease or discontinue this treatment. Complications included transient elevation of intraocular pressure in 4 patients and persistent mild ptosis in 2 patients. CONCLUSIONS: We have demonstrated that posterior sub-Tenon's triamcinolone injection significantly decreases cystoid macular oedema, with a corresponding increase in visual acuity, in patients with posterior uveitis. Systemic immunosuppression may be reduced or discontinued with the avoidance of associated systemic side effects, and the technique has a high level of patient acceptability. PMID- 9850265 TI - The correlation of the visual field with scanning laser ophthalmoscope measurements in glaucoma. AB - PURPOSE: To assess the relationship between Humphrey visual field data and optic disc topographical data collected by the Heidelberg Retina Tomograph (HRT) scanning laser ophthalmoscope in chronic glaucoma patients. METHODS: The mean deviation (MD) and corrected pattern standard deviation (CPSD) from Humphrey visual fields of 106 eyes of 106 patients with glaucoma were analysed for correlation with the multiple topographical measures calculated by the HRT. RESULTS: Significant correlations were found between MD of the visual field and several optic disc measurements. These included neuroretinal rim volume, mean nerve fibre layer thickness and cross-sectional area, and the cup shape measure. CPSD correlated significantly only with mean nerve fibre layer cross-sectional area. This pattern was common to the whole circumference of the disc with the exception of the directly temporal segment. CONCLUSION: Optic disc topography performed by HRT reflects the optic disc pathology in correlation with perimetry. PMID- 9850266 TI - Spatial and temporal visual filtering in patients with glaucoma and ocular hypertension. AB - PURPOSE: To investigate the spatial and temporal response characteristics in patients with glaucoma and ocular hypertension (OHT), in order to demonstrate the earliest specific measure of vision loss and to elucidate the precise mechanism of early glaucomatous damage. METHODS: The Open Optical View System was used to assess changes in spatial and temporal vision in patients with clearly defined glaucoma (30 eyes), patients with OHT (30 eyes) and 30 eye-matched normal control eyes. The psychophysical methods applied were developed by Barber and Ruddock, who showed that the responses called ST1 and ST2 have similar characteristics to those found in the parvocellular and magnocellular pathway respectively. The measurement of spatial and temporal responses rests upon the measurement of a target that moves across a background, itself modulated spatially (gratings) and temporally (flicker). RESULTS: The results reported, taken together with the current data on retinal ganglion cell function, reveal damage to both the parvocellular and magnocellular pathway in patients with glaucoma. In the OHT groups only marginal changes were found in the parvocellular response, but significant changes in the magnocellular response. In both OHT and glaucoma, the temporal processing shows greatest abnormalities at a small number of low flicker frequencies. CONCLUSION: We provide psychophysical evidence of the detectable ganglion cell damage in patients with OHT who remain normal on testing with automated perimetry. The psychophysical method chosen (which measures the function of specific sub-divisions of the visual pathway) may prove useful in screening for glaucomatous damage. PMID- 9850267 TI - Lack of cross-reactive allergic response to brimonidine in patients with known apraclonidine allergy. AB - PURPOSE: To assess the allergenic potential of topical brimonidine 0.2% in patients shown to be allergic to apraclonidine 0.5%. METHODS: Eighteen glaucoma patients who developed allergy to apraclonidine were enrolled. Allergy was graded and documented photographically. Apraclonidine was then discontinued until the allergy resolved. Grading and photography were repeated, following which treatment with brimonidine was initiated. Patients were examined 1 h after the initial drop of brimonidine, twice weekly for 6 weeks, and at 3 monthly intervals thereafter. All other anti-glaucoma medications were continued. RESULTS: Mean patient age was 66.3 +/- 14.9 years (range 33.5-89.3 years). The mean time to apraclonidine allergy from initial exposure was 12.9 +/- 12.9 months (range 2.1 46.8 months). For the 10 patients rechallenged with apraclonidine, the mean time to allergy was 13.6 +/- 10.2 days (range 3-32 days). Mean duration of brimonidine therapy was 140.7 +/- 66.2 days (range 14-286 days), with 11 patients reaching at least 150 days of follow-up. Two patients developed symptomatic brimonidine allergy at 138 and 201 days respectively. For those patients who did not develop symptoms of allergy to brimonidine, there was no significant change in any of the parameters by which allergy was graded throughout the study. CONCLUSIONS: Patients with known apraclonidine allergy do not develop an early allergic response to brimonidine. These data rule against a cross-reactive allergic response between these two alpha 2-adrenoreceptor agonists. PMID- 9850268 TI - Delayed post-operative use of 5-fluorouracil as an adjunct in medically uncontrolled open angle glaucoma. AB - PURPOSE: The current study was performed to evaluate the efficacy and safety of late post-operative use of subconjunctival 5-fluorouracil (5-FU) with ocular massage (OM) after trabeculectomy with or without intraoperative mitomycin C (MMC) in patients with medically uncontrolled primary open angle glaucoma (POAG). METHODS: Initial trabeculectomy was performed in 60 eyes of 60 consecutive patients with medically uncontrolled POAG. Thirty eyes (group 1) were randomly assigned to intraoperative MMC (0.2 mg/ml for 3 min) and 30 (group 2) were randomised to standard trabeculectomy. During the first 3 months post-operatively OM and subconjunctival injections of 5-FU (5 mg in 0.5 ml of balanced salt solution) were performed in both groups every time the clinical evaluation suggested imminent bleb failure. Suture lysis was not performed in any patient. RESULTS: Follow-up ranged from 14 to 53 months (mean 30.17, SD 9.23) in trabeculectomy + MMC treated eyes and from 6 to 54 months (mean 27.37, SD 10.83) in trabeculectomy treated eyes. During the first 3 months of follow-up OM and subconjunctival injections of 5-FU were performed in 14 cases in group 1 and in 18 cases in group 2 (p = NS). A positive response to OM was obtained in 14 of 14 eyes and in 14 of 18 eyes in group 1 and in group 2, respectively (p = 0.042). The difference in post-operative mean IOP between the two groups was statistically significant at each time interval studied (p < 0.001). Success (complete or qualified) was achieved in 50 of 60 eyes (83.33%): 28 (93.3%) in the trabeculectomy + MMC treated group and 22 (73.3%) in the trabeculectomy treated group (p = 0.039). Among the complications seen, the incidence of bleb fibrosis was higher in group 2 (p = 0.0026). By means of post-operative treatment four nonrandomised subgroups were identified: intraoperative MMC + post-operative 5 FU, intraoperative MMC + no post-operative 5-FU, no intraoperative MMC + post operative 5-FU, no intraoperative MMC + no post-operative 5-FU. The eyes treated with intraoperative MMC and post-operative 5-FU had a better long-term (48 months) cumulative probability of success (100%); treatment with intraoperative MMC or post-operative 5-FU alone was followed by a success rate of 87.1% and 72.2%, respectively. The cumulative probability of success after only trabeculectomy was 56% (p < 0.05). One case of hypotony maculopathy was found in the subgroup treated only with intraoperative MMC. CONCLUSIONS: This study confirms the effectiveness and relative safety of delayed post-operative 5-FU treatment in patients with clinical evidence of bleb failure. Only when OM had caused a lowering of IOP were late subconjunctival injections of 5-FU followed by good control of IOP. The use of intraoperative MMC may ensure a greater IOP decrease after OM. PMID- 9850269 TI - Correlation between the orbital and intraocular portions of the optic nerve in glaucomatous and ocular hypertensive eyes. AB - BACKGROUND/PURPOSE: It has recently been reported that the retrobulbar optic nerve diameter (OND) and cross-sectional area (ONCSA) are reduced in glaucoma. This study was performed to investigate the correlation between the orbital and intraocular portions of the optic nerve. METHODS: One eye of 20 volunteers (16 glaucoma subjects, 4 ocular hypertension subjects) underwent optic disc analysis using Heidelberg retinal tomography, and echographic measurements of the retrobulbar optic nerve. RESULTS: The male-to-female ratio was 6.5:3.5, and the mean age of our sample (+/- SD) was 62.25 +/- 13.7 years. Orbital optic nerve diameter and cross-sectional area correlated significantly and positively with the neuroretinal rim area (Spearman's rank correlation coefficient; OND: rs = 0.488, p = 0.0336; ONCSA: rs = 0.619, p = 0.0079), but not with any other topographical disc data. The retrobulbar optic nerve cross-sectional area-to-disc area ratio (ONCSA/D) was found to have a significant negative correlation with the cup area/disc area ratio (simple regression analysis; r = -3.948, p = 0.046), and a statistically demonstrable positive correlation with the neuroretinal rim area/disc area ratio (r = 0.451, p = 0.046). CONCLUSION: The results of this study indicate that orbital optic nerve dimensions are a reflection of the neuroretinal rim area of the optic disc. Echographic measurements of the retrobulbar nerve may be additive to the traditional triad of raised intraocular pressure, field defects and glaucomatous optic neuropathy that suggests a diagnosis of glaucoma. PMID- 9850270 TI - A comparison of skin storage methods for oculoplastic surgery. AB - PURPOSE: To assess the level of contamination of full-thickness skin grafts stored with or without an antibiotic cover. METHODS: Full-thickness skin grafts were harvested from 40 bilateral upper lid blepharoplasties. Before surgery the face was sterilised, the head of the patient was packed with sterile, single-use surgical drapes and the whole face was left exposed. The harvested full-thickness skin grafts were conserved in sterile containers at 4 degrees C for 6 days, rolled in gauze moistened with either 4 ml of sterile saline solution (group I) or with 4 ml of gentamicin solution (2 mg/ml) (group II). The degree of contamination, expressed in colony forming units (CFU), was evaluated on days 2, 3, 4, 5 and 6. Identification of the microorganisms was done to species level following standard procedures and commercial methods. RESULTS: In group I 2 grafts (5%) were negative during the whole observation period while the other 38 grafts (95%) presented a degree of contamination ranging from 10(2) to 10(4) CFU. Microorganisms isolated were: Staphylococcus epidermidis (24 cases), Staphylococcus aureus (5 cases), Staphylococcus saprophyticus (2 cases), Pseudomonas aeruginosa (4 cases), Serratia liquefaciens (1 case) and Klebsiella oxytoca (2 cases). In group II, 26 grafts (65%) were negative during the whole observation time while in 14 cases (35%) a few colonies (3 to 6) of Candida albicans were isolated on day 2 and remained constant in number for the whole observation time. CONCLUSIONS: The storage of full-thickness skin graft with an antibiotic cover is more reliable than the storage of full-thickness skin graft without an antibiotic cover. PMID- 9850271 TI - Effect of bile acids on fibroblast proliferation and viability. AB - PURPOSE: To examine the in vitro effects of bile acids and salts on the viability, growth and morphology of human Tenon's capsule fibroblasts. METHODS: Human Tenon's capsule fibroblasts were grown from explants in culture and used between passages 3 and 5. Fibroblasts (n = 3) were treated in triplicate with deoxycholic acid (DA), sodium deoxycholate (NaD) and cholic acid (CA) at concentrations between 50 microM and 1 mM. Cell viability and growth were assessed with trypan blue staining and haemocytometer counting, and a colorimetric (MTT) assay. Morphology was assessed with light microscopy and cresyl violet staining. RESULTS: A dose-dependent decrease in viability was observed following bile acid treatment (24 and 48 h) where the effects of DA > NaD > CA. The LD50 values at 48 h for DA, NaD and CA were 300 microM, 400 microM and 720 microM respectively. Cell growth following bile acid treatment was reduced compared with controls. Fibroblasts treated with bile acids displayed a loss of normal spindle-shaped morphology and multiple processes, some with varicosities, extended from many cells. These changes were dose-dependent, and at higher concentrations cells became detached and were non-viable (trypan-blue positive). Extensive cell death was observed in cultures treated with DA 400 microM, NaD 500 microM and CA 800 microM. CONCLUSIONS: Lower doses of bile acids and salts reduced in vitro fibroblast growth associated with morphological changes; higher doses induced fibroblast cytotoxicity. These observations suggest that bile acids and salts, in particular DA, may be useful in regulating wound fibrosis following trabeculectomy surgery. PMID- 9850272 TI - New generation hydroxypropyl methylcellulose (HPMC-Ophtal H) for intraocular surgery: a confocal laser scanning microscopy study. AB - PURPOSE: To determine the abundance and variety of particulate debris in new generation hydroxypropyl methylcellulose (HPMC) samples. METHOD: Three samples of sterile HPMC-Ophtal H were examined using a confocal laser scanning microscope, scanning electron microscope and mass spectrometry. RESULTS: Confocal laser scanning microscopy of the samples showed that particulate matter in HPMC was less than 30 microns in size and at a density of less than 1000 particles per millilitre. No discernible details were seen by the scanning electron microscope, and mass spectrometry did not identify and bands. CONCLUSION: Improved methods of preparation including filtration may be responsible for decreased particulate debris in new generation HPMC. PMID- 9850273 TI - Magnetic sensor data acquisition for three-dimensional ultrasound of the orbit. AB - PURPOSE: Three-dimensional (3D) B-mode ultrasound imaging is now possible using a magnetic sensor data acquisition system to localise data points, allowing free hand scanning. We report the first use of this freehand scanning system for insonation of the orbit. METHODS: We insonated 10 orbits in 5 volunteers (all men; mean age 37 years) with an Acuson 128XP ultrasound system and a 7.5 MHz transducer. Power-Doppler was used to image vascular structures. Data were acquired using a new magnetic sensor system with detectors mounted on the transducer. Free-hand scanning was done using sweeping or fan-like and linear movements over 20-30 s to cover the orbit, with retrospective measurements of orbital structures. A 3D reconstruction was performed via an external workstation. RESULTS: All orbits were adequately imaged by one data acquisition. Selected intraorbital structures were identified and their course followed, including optic nerve and central vessels, as well as short and long ciliary vessels. The mean transverse area of the optic nerve was 5.6 +/- 1.1 mm2; the diameter of the optic nerve was 3.0 +/- 0.3 mm. The lateral rectus muscle was clearly seen, with a mean diameter of 4.9 +/- 0.3 mm, at the level of optic nerve head. CONCLUSION: Three-dimensional ultrasound of the orbit allows imaging of the location, course and relationships of intraorbital vessels, the optic nerve and the lateral rectus muscles with one data acquisition, as well as quantitative measurements not possible with 2D images alone. This method allows rapid, free hand data acquisition, with a multitude of potential clinical applications. PMID- 9850275 TI - Collagen content and types in trachomatous conjunctivitis. AB - PURPOSE: To study alterations in conjunctival collagen in the conjunctiva of patients with active trachoma. METHODS: We studied conjunctival biopsy specimens obtained from nine subjects with active trachoma and from four control subjects. We used immunohistochemical techniques and a panel of monoclonal and polyclonal antibodies directed against types I, III, IV and V collagen. RESULTS: In normal conjunctiva, the staining for types I and III collagen was localised to the substantia propria. Type IV collagen was located in the epithelial and capillary endothelial basement membranes. The staining for type V collagen was absent. In trachoma biopsy specimens, staining for types I and III collagen showed collagen fibrils among epithelial cells, patchy increase in staining intensity in the upper stroma, and thicker and irregularly arranged collagen fibrils in the substantia propria. Staining for type IV collagen showed irregularly thickened epithelial basement membrane. Staining for type V collagen showed patchy staining in the upper substantia propria; it was also noted in the cytoplasm of fibroblasts, in the walls of blood vessels in the substantia propria, and in the walls of accessory lacrimal glands. CONCLUSIONS: Our data indicate new type V collagen formation, and increased types I, III and IV collagen content, in the conjunctiva from patients with active trachoma. PMID- 9850274 TI - Tenascin-cytotactin (TN-C) variants in pseudophakic/aphakic bullous keratopathy corneas. AB - PURPOSE: To examine pseudophakic/aphakic bullous keratopathy (PBK/ABK) human corneas for patterns of expression of tenascincytotactin (TN-C) variants known to mediate specific cellular functions, viz. anti-adhesion (high molecular mass (M(r))) and adhesion (low/intermediate M(r)). METHODS: PBK/ABK corneas were selected to encompass only those with bullae and without inflammation, scarring or neovascularisation. Serial sections from these and normal corneas were labelled with antibodies BC-4 (recognising all TN-C variants) and BC-2 (specific for the high M(r) TN-C variant). Bound antibody was revealed with an avidin biotin peroxidase technique. In a given pair of corneal sections, positivity with BC-4 but not BC-2 indicates localisation of low/ intermediate M(r) TN-C variants and absence of the high M(r) TN-C variant. BC-2 identifies the high M(r) variant. RESULTS: There was no immunostaining with either BC-2 or BC-4 in normal corneas except at the corneoscleral interface, where both BC-2 and BC-4 were immunolocalised. In PBK/ABK corneas, BC-2 staining was seen in 5 of 13 corneas and was restricted mainly to epithelial basement membrane (BM) overlying bullae. BC-2 did not label the stroma. BC-4 immunostaining was present in all PBK/ABK corneas and was localised in epithelial BM, both epithelial and stromal borders of bullae, pannus, endothelial BM and in oedematous stromal regions. CONCLUSIONS: TN-C variants are differentially expressed in PBK/ABK corneas. The high M(r) variant is restricted mainly to epithelial BM overlying bullae, while low/intermediate M(r) variants occur in epithelial BM, both epithelial and stromal borders of bullae, and in pannus. Given the in vitro functions of TN-C, a role for promoting epithelial dehiscence and reattachment to the substratum in PBK/ABK corneas by high and low/intermediate M(r) variants respectively is likely. PMID- 9850276 TI - Unusual macular findings in a known choroideremia carrier. PMID- 9850277 TI - Darkening of eyelashes in a patient treated with latanoprost. PMID- 9850278 TI - Corneal endothelial precipitates in HIV- and CMV-positive patients without concomitant ocular disease. PMID- 9850279 TI - Pedunculated episcleral choristoma without ocular involvement. PMID- 9850280 TI - Conjunctival lymphangioma. A can of worms? PMID- 9850281 TI - Diagnosis of Theodore's superior limbic keratoconjunctivitis. PMID- 9850282 TI - The National Survey of Local Anaesthesia for Ocular Surgery. PMID- 9850283 TI - Results of primary retinal detachment surgery: a prospective audit. PMID- 9850284 TI - Results of primary retinal detachment surgery: a prospective audit. PMID- 9850285 TI - Relationship of diabetic microvascular complications to outcome in panretinal photocoagulation treatment of proliferative diabetic retinopathy. PMID- 9850286 TI - Prenatal diagnosis: the way ahead? PMID- 9850287 TI - The future of gene therapy in obstetrics and gynaecology. PMID- 9850288 TI - Making the most of MEDLINE. AB - The MEDLINE database is firmly established as an essential tool in medical research and practice, and all doctors should develop MEDLINE search skills. This article reviews key elements of the database, looking particularly at areas of common error, and the options available when fine-tuning a search. A better understanding of MEDLINE should lead to more effective use of this invaluable resource. PMID- 9850289 TI - Evidence-based medicine as a tool. AB - Clinicians need current best evidence to help them treat their patients optimally. It is impossible for them to read all the relevant clinical literature and their reading time decreases after completion of medical school. The practice of evidence-based medicine helps clinicians meet these challenges. PMID- 9850290 TI - Temperature charts: a time for change? AB - Today temperature charts are used at the bedside in hospitals throughout the world. The diagnosis, investigation and treatment of patients is based on the information recorded on these charts but without a knowledge of clinical thermometry the current chart is open to misinterpretation. This article will review the origins of the current chart and discuss proposals for a new chart. PMID- 9850291 TI - An inexpensive and easily constructed laparoscopic simulator. AB - In this short article the authors describe a simple yet effective method to train surgical trainees in minimally invasive surgery. This method will allow trainees to freely practice and acquire the necessary basic skills required to both competently assist and perform laparoscopic surgery. PMID- 9850292 TI - Shingles: a review of diagnosis and management. AB - Herpes zoster or shingles results from reactivation of varicella zoster virus previously dormant in cells of the dorsal root ganglion. The incidence of shingles increases with age and immunosuppression. Guidelines for managing shingles are now available and implementation, with the emphasis on early treatment, may reduce the severity of a shingles attack and reduce the incidence of complications. PMID- 9850293 TI - Use of intracavernosal alprostadil in erectile dysfunction. AB - Alprostadil (prostaglandin E1) is the most widely used, licensed vasoactive agent for intracorporeal injection in the investigation and treatment of erectile dysfunction. It has a good side-effect profile and is effective in between 70 and 80% of patients. PMID- 9850294 TI - An approach to switching patients from baclofen to tizanidine. AB - With the introduction of the antispasticity agent, tizanidine hydrochloride (Zanaflex), physicians have requested information about the optimal way to switch appropriate patients from baclofen to tizanidine. A group of neurologists and rehabilitation specialists with a particular interest in spasticity was therefore asked to draw up a suitable approach to changing treatment. PMID- 9850295 TI - Imaging the painful shoulder: an update. AB - The shoulder is a complex, inherently unstable joint which may be afflicted by a variety of lesions. Although newer imaging modalities, such as computed tomography, and magnetic resonance imaging and arthrography, can be used to assess the shoulder, plain X-ray is still recommended for initial imaging. Current applications of diagnostic imaging in patients presenting with painful shoulders are reviewed. PMID- 9850296 TI - Endovascular repair for abdominal aortic aneurysms. AB - Endovascular exclusion could allow safe and successful treatment of abdominal aortic aneurysms. It may also be possible to use this technique in patients classed as high risk for conventional open repair. However, it is a new technique which has not been validated in a proper clinical trial. This review aims to outline current thinking in this field and the basis of the procedure. PMID- 9850297 TI - Bullectomy for chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease is a progressive, disabling condition which leads to poor quality of life and limited survival. Over the years, a variety of surgical procedures have been used to achieve symptomatic improvement in selected patients. This article gives a historical account of these interventions and looks at current surgical options, both for primary treatment and management of complications. PMID- 9850298 TI - Acute myocardial infarction: failed thrombolysis revisited. AB - The prognosis following acute myocardial infarction is dependent upon the presence of adequate blood flow in the infarct-related vessel. All too often, this is not achieved by thrombolytic therapy ('failed thrombolysis'). Diagnosis of failed thrombolysis is difficult and the optimum management is currently unclear. PMID- 9850299 TI - Interpreting reported health-care costs. AB - This short paper describes the different categories of health-care costs and methodological considerations in cost measurement. It provides simple advice on avoiding some of the common pitfalls in cost interpretation and should help readers in assessing the usefulness of reported costs. PMID- 9850302 TI - A national trainee advisory network: a proposal. AB - Trainees should have a well-defined and consistent communication conduit to the authorities involved with their training. It is important that trainees are involved in the evolution of structured training programmes and for training authorities to receive feedback from individuals in each specialty. This paper proposes a structure for trainee representation on regional and national training authorities to facilitate an advisory network. PMID- 9850301 TI - Irbesartan treatment in hypertension. AB - Angiotensin II receptor antagonists lower blood pressure by blocking the final step in the renin pathway, whereas ACE inhibitors reduce angiotensin II production. ACE inhibitors also block kinin degradation, which may alter both efficacy and side-effects. Irbesartan, one of the newer receptor antagonists, has confirmed their tolerability and shown similar dose-related efficacy to other major classes. PMID- 9850303 TI - Higher level practice in nursing: a prerequisite for nurse consultants? AB - The Prime Minister has stated that he wishes nurse consultant posts to be introduced in the NHS. The UKCC is currently consulting on a framework for recognizing nurses who are working at a higher level of practice, and this work could form the professional standards infrastructure for those seeking a nurse consultant post. PMID- 9850304 TI - Fingertip metastasis presenting with a history of trauma. PMID- 9850305 TI - Cost-benefit analysis of spinal cord stimulation for intractable angina pectoris: a win-win scenario? PMID- 9850306 TI - Benign prostatic hyperplasia and hypertension. PMID- 9850307 TI - Anaesthesia in orthogeriatric rehabilitation. PMID- 9850308 TI - Nuclear medicine and endocrinology. PMID- 9850309 TI - An insight into the options available for equity investment. PMID- 9850310 TI - Sevoflurane induction in acute airway obstruction. PMID- 9850311 TI - Treacher Collins syndrome: from linkage to prenatal testing. PMID- 9850312 TI - Impact of phenotype on treatment and prognosis of laryngeal malignancies. AB - An overview of the impact of the phenotype on treatment and prognosis of different laryngeal malignancies is presented. PMID- 9850313 TI - Toynbee Memorial Lecture 1997. Middle ear mechanics in normal, diseased and reconstructed ears. AB - A review of the structure-function relationships in normal, diseased and reconstructed middle ears is presented. Variables used to describe the system are sound pressure, volume velocity and acoustic impedance. We discuss the following: (1) Sound can be transmitted from the ear canal to the cochlea via two mechanisms: the tympanoossicular system (ossicular coupling) and direct acoustic stimulation of the oval and round windows (acoustic coupling). In the normal ear, middle-ear pressure gain, which is the result of ossicular coupling, is frequency dependent and smaller than generally believed. Acoustic coupling is negligibly small in normal ears, but can play a significant role in some diseased and reconstructed ears. (2) The severity of conductive hearing loss due to middle-ear disease or after tympanoplasty surgery can be predicted by the degree to which ossicular coupling, acoustic coupling, and stapes-cochlear input impedance are compromised. Such analyses are used to explain the air-bone gaps associated with lesions such as ossicular interruption, ossicular fixation and tympanic membrane perforation. (3) With type IV and V tympanoplasty, hearing is determined solely by acoustic coupling. A quantitative analysis of structure-function relationships can both explain the wide range of observed post-operative hearing results and suggest surgical guidelines in order to optimize the post-operative results. (4) In tympanoplasty types I, II and III, the hearing result depends on the efficacy of the reconstructed tympanic membrane, the efficacy of the reconstructed ossicular chain and adequacy of middle-ear aeration. Currently, our knowledge of the mechanics of these three factors is incomplete. The mechanics of mastoidectomy and stapedectomy are also discussed. PMID- 9850314 TI - Grommet insertion in children: a survey of parental perceptions. AB - Grommet insertion is a widely accepted method of treatment of glue ear in children. There have been questions raised over the last few years about the indications for grommets and whether assessing the hearing alone is an efficient outcome measure. Parental pressure accounts for one of the factors that is taken into consideration when the decision to insert grommets for glue ear is made. In this paper, a prospective questionnaire is used to investigate the parental perceptions of the effectiveness of grommet insertion in children, focusing on alternative outcome measures such as general health, language, and social skills. The results of this survey suggest that grommet insertion causes improvement in many factors other than hearing and this seems to account for the parental pressure for siblings to have grommet insertion. PMID- 9850315 TI - Surgery for acquired cholesteatoma in children: long-term results and recurrence of cholesteatoma. AB - The aim of the study was to evaluate the long-term results after surgery for acquired cholesteatoma in children and to contribute to the search for predictors of recurrence. During a 15-year period, 114 children underwent surgery. The patients were re-evaluated with a median observation time of 5.8 years. At the last re-evaluation 85 per cent of the ears were dry with an intact drum. Recurrence of cholesteatoma developed in 27 ears. The cumulated total recurrence rate was 24 per cent using the incidence rate calculation, applying Kaplan-Meier survival analysis the corresponding recurrence was 33 per cent. Recurrent disease occurred significantly more frequently in children younger than eight years, with a negative pre-operative Valsalva, with ossicular resorption and with large cholesteatomas. In conclusion, young children with poor Eustachian tube function and a large cholesteatoma with erosion of the ossicular chain, are at special risk of recurrence and should be observed for several years after surgery. PMID- 9850316 TI - Expression of transforming growth factor-alpha (TGF-alpha) in cholesteatoma. AB - This study aims at elucidating the role of cytokines in the mechanism of proliferation of cholesteatoma epithelium by investigating the mode of expression of epidermal growth factors, such as TGF-alpha. The subjects of this study were patients who had undergone operation for middle ear cholesteatoma. Skins of the bone region of the external ear canal (normal skin) of the same patients were used as the negative control. The mode of expression of TGF-alpha was studied by immunohistochemistry and in situ hybridization. In the immunohistochemical study, there were no conspicuous differences observed between cholesteatoma tissues and normal skin. After in situ hybridization, expression of TGF-alpha mRNA was mainly observed in the epidermal basal cell layer in the normal skin, while in the cholesteatoma epidermis with severe inflammatory cell infiltration, expression of TGF-alpha mRNA was observed up to layers superior to the basal cell layer. The expression of TGF-alpha mRNA is greatly affected by subepithelial connective tissue, strongly suggesting involvement of paracrine regulation in proliferation of cholesteatoma epithelium. PMID- 9850317 TI - Epithelial atypia and squamous metaplasia in nasal polyps. AB - Fifteen routine nasal polypectomy specimens submitted to our laboratory over an eight-month period were noted to show marked atypia within areas of squamous metaplasia to a degree suggestive of dysplasia. Reviewing the literature revealed little published work in this area, especially recently. Further nasal polyp cases from this, and a similar previous time period, were re-examined, and possible causes for these changes discussed. PMID- 9850318 TI - Endoscopic resection of inverted papilloma of the nose and paranasal sinuses. AB - The results of a retrospective study of 22 patients with inverted papillomas resected by the endoscopic approach are presented with a follow-up of 33 to 96 months. Twenty-one patients had unilateral disease and one patient had bilateral involvement. None of the patients had orbital or cranial extension. One patient had synchronous carcinoma in situ. Eight patients had undergone previous surgical procedures. Following endoscopic surgery, six patients had residual disease requiring further revisions. Three of these six patients eventually required excision via limited external approaches. No patient required lateral rhinotomy or mid-facial degloving procedures. No complication occurred in any of the patients. The advantages of endoscopic surgery include precise determination of tumour extent, preservation of normal mucosa and bony structures and avoidance of external scars. Close endoscopic follow-up is mandatory to ensure early recognition and treatment of recurrent disease. Although the endoscopic approach is gaining popularity for the treatment of inverted papilloma, indiscriminate application may result in a high recurrence rate. The endoscopic approach should be performed by experienced surgeons and restricted to carefully selected patients with nasal, ethmoidal and limited maxillary disease. More extensive disease should be managed by radical external approaches or by combining endoscopic with limited external approaches. PMID- 9850319 TI - Management of subglottic haemangioma. AB - Two contrasting methods of treatment for paediatric subglottic haemangioma are presented. Dr Hoeve describes his results using intralesional steroids and short term intubation, and Dr Froehlich discusses his outcomes with open surgical excision. Mr Bailey then reviews the currently available therapeutic options with special reference to each of the above techniques, and suggests an optimum plan of management given our current state of knowledge. The conclusion is that very small haemangiomas may not require treatment, or may be amenable to CO2 laser vaporization. Medium-sized lesions seem suitable for intra-lesional steroids and intubation, but large ones are probably best managed by primary submucous resection. Very large haemangiomas, and especially those which are circumferential or in which magnetic resonance imaging (MRI) shows extension down into the trachea and/or through the tracheal wall into the surrounding tissues, may be more safely dealt with by performing a tracheostomy and awaiting spontaneous involution. PMID- 9850320 TI - Bilateral sudden sensorineural hearing loss following non-otologic surgery. AB - Hearing loss as a complication of non-otological surgery is rare. The majority of described cases have followed cardiac bypass surgery. To date there have been only 25 reported cases of non-otological surgery associated hearing loss (NOSAHL) which have not involved cardiopulmonary bypass. Only two cases of bilateral NOSAHL have been previously described, and neither resulted in permanent severe bilateral hearing deficit. We describe a case of permanent bilateral severe hearing loss following metatarsal pinning in a patient with pre-existing non operated otosclerosis. PMID- 9850321 TI - Aberrant jugular bulb vein obstructing approach to intracanalicular vestibular schwannoma. AB - In cases of jugular bulb anomalies, such as a high jugular bulb, troublesome bleeding may occur during surgery. We report an unusual case with a vestibular schwannoma, in which we recognized an additional branch arising from the jugular bulb. Three-dimensional computed tomography (3-D CT) revealed this anomaly beforehand, enabling us to avert excessive bleeding upon resection of the tumour. The abnormal vein was thought to be a remnant of the petrosquamosal sinus in the embryonic stage. PMID- 9850322 TI - An unusual presentation of a nasal septal abscess. AB - Nasal septal abscess is a rare complication of septal haematoma. Nasal obstruction and, less frequently, pain are the usual presenting features. We report a case of a nasal septal abscess in a 21-year-old female patient who developed a naso-oral fistula. To our knowledge this is the first report of such an unusual presentation of a septal abscess. The aetiology, pathogenesis and management of septal abscesses are discussed. PMID- 9850323 TI - Cavernous sinus syndrome caused by a primary paranasal sinus non-Hodgkin's lymphoma. AB - Although uncommon, non-Hodgkin's lymphomas occasionally arise from the nose and paranasal sinuses. Rarely, they may invade into the cavernous sinus and produce signs and symptoms that characteristically include unilateral ophthalmoplegia, sensation loss in the distribution of the ophthalmic and other divisions of the trigeminal nerve, sympathetic nerve paralysis and proptosis. In this report, we present a case of cavernous sinus syndrome (CSS) caused by infiltration of non Hodgkin's lymphoma from the adjacent paranasal sinuses and address issues regarding its diagnosis and treatment. PMID- 9850324 TI - A Le Fort I osteotomy approach to lateral sphenoid sinus encephalocoeles. AB - Meningo-encephalocoeles of the skull base may present as spontaneous cerebrospinal fluid rhinorrhoea or acute meningitis. Previous approaches to midline skull base lesions have been either intracranial, via a craniotomy, or by transfacial or endoscopic extracranial approaches. This paper presents an alternative approach to lateral sphenoid sinus encephalocoeles through a Le Fort I osteotomy approach. PMID- 9850325 TI - Tuberculosis of the pyriform fossa--a rare entity. AB - Tuberculosis of the pharynx is less common than tuberculosis of the larynx. We present a rare case of tuberculosis of the pyriform fossa which clinically masqueraded as a malignancy. Our patient showed a prompt improvement in symptoms after commencing antitubercular treatment. PMID- 9850327 TI - Replacing the combitube by an endotracheal tube using a fibre-optic bronchoscope during spontaneous ventilation. AB - We present a case of microlaryngoscopy in a patient with an unexpectedly difficult airway. The airway was managed by using an oesophageal-tracheal Combitube (Kendall-Sheridan, Argyle, NY) (ETC) and a fibre-optic bronchoscope (Pentax-Japan-5 mm). PMID- 9850326 TI - A megalith of the parotid salivary gland. AB - We present a rare case of a giant intra-parotid calculus in a 64-year-old man with recurrent parotitis. The literature is also reviewed. PMID- 9850328 TI - Have we got the full picture? AB - A 59-year-old man with long-standing chronic obstructive airways disease (COPD), became progressively dyspnoeic, and repeatedly blacked-out during forced expiration. Spirometry suggested the possibility of large airways obstruction, and failing to respond to aggressive bronchodilator and steroid therapy, the patient was labelled as being non-compliant. Finally, he was assessed by an otolaryngologist and a cause for upper airway obstruction was suspected. Bronchoscopy and computed tomography (CT) scanning demonstrated tracheomalacia and the patient underwent resection of this segment of abnormal trachea. Tracheomalacia, although rare, results from the substitution of cartilage with fibrous tissue, leading to severe airway compromise. This case emphasizes the fact that although many conditions are uncommon, the total incidence of rare conditions is surprisingly high, and that care needs to be taken at all times in the management of 'labelled' patients with chronic illness, in order not to overlook such life-threatening diagnoses. PMID- 9850329 TI - Encephalocoele as a complication of intranasal sinus surgery: optimal evaluation with magnetic resonance imaging. AB - We report a case of post-operative frontal basal encephalocoele evaluated using a new magnetic resonance imaging (MRI) sequence, fast inversion recovery for myelin suppression (FIRMS). FIRMS was developed to enhance the differentiation between grey and white matter. In this case, the sequence was beneficial in distinguishing the encephalocoele from adjacent nasal mucosa and secretions. PMID- 9850330 TI - Mycotic aneurysm of the external carotid artery. AB - Mycotic aneurysms of the extracranial carotid arteries are extremely rare. A case is reported of a false aneurysm of the left external carotid artery. This developed secondary to cervical lymphadenitis which did not settle with high dose antibiotic therapy. The diagnosis was made on investigation with carotid doppler ultrasound and confirmed with computerized tomography. Digital subtraction angiography was performed to highlight the vascular anatomy. In addition percutaneous balloon catheter control of blood flow in the external carotid artery was used as an adjunct to surgical management. PMID- 9850331 TI - Power Doppler imaging findings in multilocular giant parathyroid adenoma which caused hypercalcaemic crisis. AB - Radiological findings including ultrasonography, computed tomography (CT) and Tc 99m sestamibi scintigraphy of a patient with multilocular giant parathyroid adenoma which caused hypercalcaemic crisis are presented. The location of the tumour by grey scale sonography, CT and Tc-99m sestamibi scintigraphy was not certain because the tumour was uncommon in shape, location, size and internal structure. Whereas, increased flow in the solid portion of the mass was demonstrated on power Doppler sonography, which proved to reflect abundant vessels in the adenoma in pathological findings. PMID- 9850332 TI - Actinomycosis of the middle ear. AB - Another case of actinomycosis of the ear is described in a nine-year-old boy, drawing attention to the increasing incidence of diseases of the ear considered to be rare. The patient presented with the clinical signs of chronic purulent otitis media, not responding to conservative treatment. After the diagnosis of actinomycosis was established the patient was treated by surgery and long-term antibiotic medication, responding favourably. PMID- 9850333 TI - Angiolymphoid hyperplasia with eosinophilia of the parapharyngeal space. AB - Angiolymphoid hyperplasia with eosinophilia (ALHE) is an uncommon benign condition characterized by cutaneous nodules with a predeliction for the head and neck region. Extracutaneous involvement is rare. We report a 44-year-old woman who had a large submucosal ALHE tumour in the parapharyngeal space. Our patient is of interest because of the unusual, and as far as we are aware from the literature, unique site and presentation of her lesion. PMID- 9850334 TI - In cell amplification. PMID- 9850335 TI - Differences in p53 and Bcl-2 expression in relation to cell proliferation during the development of human embryos. AB - AIMS: To study the patterns of p53 and Bcl-2 expression in relation to cell proliferation during human embryogenesis in order to help elucidate their potential roles in the regulation of cell proliferation and apoptosis during morphogenesis. METHODS: Immunohistochemistry for p53, Bcl-2, and proliferating cell nuclear cell antigen (PCNA) proteins was performed, using a variety of monoclonal antibodies, on paraffin was embedded sections of tissues from 68 human embryos and fetuses of between 4 and 30 weeks gestation. RESULTS: Positive relations between sites of proliferative activity (as detected by PCNA expression) and p53 expression were found in the kidney, early developmental stages of intestine and lungs, liver, pancreas, heart, and in embryonic osteoblasts. On the other hand, positive relations between proliferative activity and Bcl-2 expression were found in the gonads, adrenal glands, in the cells of the dental lamina, hair follicles, syncytiotrophoblast, chondrocytes, and more advanced stages of intestinal development. In tissues of the central nervous system, p53 and Bcl-2 were co-expressed at the same sites but there was an inverse relation between p53/Bcl-2 expression and proliferative activity. CONCLUSIONS: These data suggest that p53 and Bcl-2 have tissue specific and stage specific functions during embryogenesis. PMID- 9850336 TI - Cytochrome P450 1B1 mRNA in the human central nervous system. AB - AIMS: To study the expression of CYP1B1 in a variety of human and rat cell lines as a means of identifying a new tool for the investigation of gene regulation. In addition, to identify the expression of cytochrome P450 1B1 (CYP1B1) in different regions of the central nervous system (CNS). METHODS: Reverse transcription polymerase chain reaction followed by cloning and sequencing were used to detect the expression of CYP1B1 in human cell lines. Poly A+ mRNA from the human spinal cord and from different brain regions was analysed using a CYP1B1 probe labelled with 32PdCTP. RESULTS: Expression of CYP1B1 was shown in a human astrocytoma cell line (MOG-G-CCM). CYP1B1 mRNA was expressed in a variety of regions of the CNS but with a distinct regional specificity. Expression was highest in the putamen. CONCLUSIONS: The expression of CYP1B1 in a human astrocytoma enables this cell line to be used in further studies of regulation and function of this gene. The demonstration that CYP1B1 mRNA is expressed in a variety of regions of the CNS suggests a role for this gene in brain and spinal cord metabolism. The regional specificity of expression might explain the focal damage of certain human neurodegenerative diseases. PMID- 9850338 TI - Monitoring of Chlamydia trachomatis infections after antibiotic treatment using RNA detection by nucleic acid sequence based amplification. AB - AIM: To investigate the value of RNA detection by nucleic acid sequence based amplification (NASBA) for the monitoring of Chlamydia trachomatis infections after antibiotic treatment. METHODS: Cervical smears (n = 97) and urine specimens (n = 61) from 25 C trachomatis positive female patients were analysed for the presence of C trachomatis 16S ribosomal RNA (rRNA) by NASBA and C trachomatis plasmid DNA by the polymerase chain reaction (PCR) before and up to five weeks after antibiotic treatment. RESULTS: Chlamydia trachomatis RNA was found in all cervical smears taken before antibiotic treatment (n = 24) and in two smears taken one week after antibiotic treatment; no C trachomatis RNA was detected after two weeks or more. In contrast, C trachomatis DNA was found in all such specimens before treatment, and 21 of 25, six of 21, and five of 20 smears were found to be positive at one, two, and three weeks after treatment, respectively. After four weeks, only one of six smears was positive, and this smear had been negative in the two preceding weeks. Of the 61 urine samples investigated, C trachomatis DNA and C trachomatis RNA were found in all before treatment (n = 15), whereas one week after treatment four of 15 were C trachomatis DNA positive and C trachomatis RNA was detected in one sample only. CONCLUSIONS: These data show that RNA detection by NASBA can be used successfully to monitor C trachomatis infections after antibiotic treatment. Furthermore, it might be possible to use urine specimens as a test of cure because neither C. trachomatis DNA or RNA could be detected two weeks or more after treatment. PMID- 9850337 TI - Chemotaxins C5a and fMLP induce release of calprotectin (leucocyte L1 protein) from polymorphonuclear cells in vitro. AB - AIMS: To determine whether the chemotaxins C5a and formyl peptide (fMLP) can stimulate the release of calprotectin, the major leucocyte protein of polymorphonuclear neutrophils (PMN). METHODS: A dose response curve for the uptake of 125I labelled rC5a and fMLP in PMN was determined by radioimmunoassay. The unlabelled chemotaxins were then incubated with PMN and the concentration of calprotectin in PMN lysates and supernatants was measured by an enzyme immunoassay. RESULTS: Both rC5a and fMLP induced release of calprotectin from PMN in a dose dependent manner as determined by a reduction in intracellular calprotectin concentration. A minimum of approximately 10% of total PMN calprotectin was retained at concentrations of 10-100 nM of rC5a and 0.1-10.0 nM of fMLP. Antibodies to C5a reduced the rC5a mediated release of calprotectin, and the fMLP antagonist N-t-Boc-MLP inhibited the fMLP induced calprotectin release. Because receptors for rC5a (CD88) and fMLP are G protein coupled and thought to be pertussis toxin sensitive, PMN were incubated with this toxin before the experiments. The toxin was found to reduce uptake of rC5a by the cells and to inhibit rC5a and fMLP mediated calprotectin release. CONCLUSIONS: rC5a and fMLP mediate release of calprotectin from PMN in vitro. This effect might be important during human infections in vivo. PMID- 9850339 TI - Human herpes virus 8 (HHV-8) in Kaposi's sarcoma: lack of association with Bcl-2 and p53 protein expression. AB - AIMS: Human herpes virus 8 (HHV-8) is the infectious agent implicated in the pathogenesis of Kaposi's sarcoma, although its mode of action is unclear. Recent work indicates that the HHV-8 genome encodes a viral Bcl-2 homologue (v-Bcl-2). The aim of this study was to explore Bcl-2 expression in Kaposi's sarcoma using a unique set of HHV-8 positive and negative cases, and to determine whether there is a relation with p53 expression. METHODS: Up to 18 specimens from 17 patients were selected. HHV-8 status was determined using nested polymerase chain reaction (PCR) to the open reading frame (ORF) 26, with further confirmation by TaqMan PCR. In addition, Bcl-2 and p53 immunohistochemistry were performed using standard protocols. RESULTS: The results suggest that Bcl-2 and p53 expression is independent of HHV-8 status. In addition, there does not appear to be a direct correlation with disease stage. CONCLUSIONS: HHV8 histopathogenesis is likely to be a multifactorial complex process, which may be mediated in part by viral genes and apoptosis regulating homologues. PMID- 9850340 TI - A novel, rapid in cell RNA amplification technique for the detection of low copy mRNA transcripts. AB - Growing interest now focuses on improvements of in situ polymerase chain reaction (PCR) technology for the detection of DNA and RNA cellular sequences. In this study, reverse transcription PCR in situ hybridisation (RT PCR-ISH) was developed and used to determine gene expression of pyruvate dehydrogenase in a cell model system, using human peripheral blood lymphocytes (PBLs). The success of in cell RNA amplification depends on the type of cell/tissue fixation, cell permeabilisation, and the efficiency of reverse transcription and cDNA amplification. This paper presents new approaches to overcome the critical aspects of fixation, permeabilisation, and reverse transcription when performing in cell RNA amplification. A novel fixative, "Permeafix", possessing fixative and permeabilisation properties, was used for cell fixation procedures. "Permeafix" obviated the need for pre-amplification proteolysis, facilitating entry of PCR reagents to target sequences within the cell. In addition, a simple on step RNA in cell amplification protocol using recombinant Thermus thermophilus (rTth) DNA polymerase, which reverse transcribes mRNA efficiently to cDNA and then catalyses cDNA amplification, was used. The value of a semi-junctional primer system for in cell gene expression studies, without the need to perform DNase digestion, is demonstrated. PMID- 9850341 TI - Same day diagnosis of Down's syndrome and sex in single cells using multiplex fluorescent PCR. AB - The major reason for prenatal diagnosis lies in the detection of trisomies, particularly trisomy 21 (Down's syndrome). Current techniques require lengthy laboratory procedures and high costs. Furthermore, diagnosis is often not possible if the sample is of small size or is contaminated. An alternative method, quantitative fluorescent polymerase chain reaction (PCR) of short tandem repeats (STRs), can also be used to diagnose trisomies and it has the advantage that a result is obtained within five to eight hours. However, this method is currently limited to relatively large amounts of sample, which restricts diagnostic confidence and value. Recently, genetic diagnosis using fluorescent PCR has been applied at the single cell level but is limited to sex or single gene defect diagnosis. This study, using quantitative multiplex fluorescent PCR, provides for the first time simultaneous diagnosis and confirmation of sex and trisomy in single cells. Two markers for chromosome 21 increase diagnostic confidence, informativeness, and confirmation. This system is rapid (five hours), reliable, and accurate and we believe that it will be more cost effective than alternative methods. The technique has direct application to preimplantation genetic diagnosis, early prenatal diagnosis, and other diagnostic systems where sample size is limited. PMID- 9850342 TI - HHV-8 is not associated with follicular dendritic cell tumours. AB - Follicular dendritic cell tumours are rare malignancies derived from the follicular dendritic cells of lymphoid follicles. These tumours have been associated with Epstein-Barr virus infections and with the hyaline vascular subtype of Castleman's disease. Because many examples of Castleman's disease have been associated with Kaposi's sarcoma associated herpes virus (HHV-8), this study uses polymerase chain reaction technology to examine five cases of follicular dendritic cell tumours for HHV-8. One of these cases had previously been documented to arise from pre-existing Castleman's disease. HHV-8 DNA was not detected in any of the follicular dendritic cell tumours examined, or in the original case of Castleman's disease. These findings suggest that HHV-8 plays no role in the aetiology of follicular dendritic cell tumours and the cause of this tumour remains obscure. PMID- 9850343 TI - Telomerase activity might persist in the human thymus throughout life. AB - Telomerase activity has been demonstrated at low levels in peripheral blood lymphocytes but at high levels in germinal centre B cells and thymocyte subpopulations. This study shows that telomerase is activated in the normal human thymus at different times of life. Telomerase activity was detected in thymic protein extracts from two newborn babies and from a 12 year old boy, as well as in extracts from two of six adult patients. The two positive cases were patients aged 54 and 66 years. These results suggest strongly that the thymus can remain functional despite involution in elderly patients. PMID- 9850344 TI - Ultrasound measurements at the proximal phalanges in healthy women and patients with hip fractures. AB - Measurements of bone mineral density (BMD) are useful for the assessment of fracture risk in osteoporosis. First prospective studies showed that quantitative ultrasound as measured at the calcaneus also predicts future hip fracture risk, independently of BMD and as accurately as BMD. The aim of this study was to compile a reference population for a new ultrasound device that determines amplitude-dependent speed of sound (AD-SOS) through the proximal phalanges of the hand and to prove its ability to distinguish between health volunteers and osteoporotic patients. In a case-control study we examined 139 healthy women aged 21-94 years and a group of 24 female patients aged 69-94 years with recent hip fractures. In the healthy reference population additional BMD measurements were performed with dual-energy X-ray absorptiometry (DXA) and quantitative ultrasound measurements at the calcaneus were carried out. In vivo precision of AD-SOS measurements through the phalanges was 0.52% CV. Simple regression analyses showed a negative correlation with age (r = -0.73, p < 0.001); modest significant correlations with BMD of the lumbar spine (r = 0.36, p < 0.001) and BMD of the femoral neck (r = 0.37, p = 0.002) as measured with DXA were shown. The comparison with another ultrasound device measuring SOS and broadband ultrasound attenuation (BUA) through the calcaneus showed correlation with SOS (r = 0.50, p < 0.001); no significant correlation was found with BUA measurements. Furthermore a dependency of AD-SOS values in anthropometric factors such as body mass index (r = 0.37, p < 0.001), height (r = 0.40, p < 0.001) and weight (r = 0.23, p < 0.05) was shown. First study results on 24 clinically diagnosed osteoporotic patients, defined as patients with recent (< 1 week) pertrochanteric or femoral neck fractures, showed a good separation between age- and sex-matched controls and osteoporotic patients (Z = -2.0 SD). Receiver operating characteristic (ROC) curves showed an area under the fitted curve of 0.83 +/- 0.06. These results are powerful for a device measuring AD-SOS through the proximal phalanges of the hand, and further prospective studies have proven the capability of phalangeal ultrasound in fracture risk assessment. PMID- 9850345 TI - A 2-year follow-up study on bone mineral density and markers of bone turnover in patients with long-standing insulin-dependent diabetes mellitus. AB - The aim of study was to evaluate, during 2-year follow-up, bone mineral density in sites with different cortical/cancellous bone ratios (lumbar spine, total body, distal site of radius) and selected markers of bone turnover (total alkaline phoshatase, osteocalcin, pyridinoline and deoxypirydinoline) in patients with long-standing insulin-dependent diabetes mellitus in comparison with healthy controls. Additionally, the influence of age, sex, smoking, duration of diabetes, the degree of metabolic control, or coexisting chronic complications of diabetes (retinopathy, incipient nephropathy, polyneuropathy) on the studied indices of bone metabolism in patients with insulin-dependent diabetes mellitus were evaluated. It was found that patients with long-standing diabetes mellitus had significantly lower bone mineral density than healthy controls (p < 0.003 in lumbar spine and p < 0.001 in total body). The incidence rate of osteopenia and osteoporosis was significantly higher in this group of patients in comparison with the controls (p < 0.005 for lumbar spine and total body and p < 0.001 for radius). In comparison with healthy subjects, diabetic patients and significantly higher, but within normal reference range, serum alkaline phosphate (p < 0.005) and osteocalcin (p < 0.05), accompanied by similar pyridinoline and not significantly increased deoxypyridinoline. Duration and metabolic control of diabetes, or the coexistence of its chronic complications, did not correlate with bone mineral density or the studied indices of bone turnover. In conclusion, diabetic osteopenia seems to be a normal bone turnover state, not influenced by the duration or degree of metabolic control of diabetes. PMID- 9850346 TI - The prevalence of low bone mineral density in Dutch perimenopausal women: the Eindhoven perimenopausal osteoporosis study. AB - The aim of this study was to estimate the prevalence of osteopenia and osteoporosis in perimenopausal women, and to assess determinants of low bone mineral density (BMD). All women born between 1941 and 1947 (aged between 46 and 54 years) living in the city of Eindhoven were invited to participate in the study: 5896 white Dutch women, representing 73% of the total number of Dutch women in this age group, were studied. Of these, 24% were using estrogen preparations and 19% had undergone hysterectomy, with or without oophorectomy. All women were interviewed and bone mineral density (BMD) of the lumbar spine was measured by dual-energy X-ray absorptiometry (DXA). Osteopenia and osteoporosis were defined according to the criteria proposed by a WHO working group. In the population studied the prevalence of osteopenia and osteoporosis was 27.3% and 4.1%, respectively. With progression from premenopause to menopause, the prevalence of osteoporosis increased from 0.4% to 12.7%, and that of osteopenia from 14.5% to 42.8%. An increased risk for low BMD (osteopenia and osteoporosis) was associated with age, menopausal status and smoking, while alcohol consumption, high body mass index (BMI) and use of estrogens had a protective effect. This study of a large population-based cohort of perimenopausal women revealed a high prevalence of low bone mass and, therefore, a higher risk for osteoporotic fractures. The data further suggest that, when issues on the long term efficacy and safety of preventive treatments are resolved, it may be possible to identify women at higher risk who are most likely to benefit from screening strategies. PMID- 9850347 TI - Comparison of quantitative ultrasound measurements in calcaneus with DXA and SXA at other skeletal sites: a population-based study on 280 children aged 11-16 years. AB - We performed ultrasound measurements (QUS) of the calcaneus in a population-based setting on 280 healthy children, aged 11-16 years, from a small urban area in southern Sweden. The results are compared with dual-energy X-ray absorptiometry (DXA) measurements in the total body, the lumbar spine and the hip, as well as single-energy X-ray absorptiometry (SXA) of the forearm. Normative data and correlations between the three different techniques were determined. We found significant correlations between QUS and age (r = 0.34-0.54), height (r = 0.13 0.56) and weight (r = 0.30-0.60), and between QUS and bone mineral density (BMD) measurements (r = 0.44-0.70). Boys increased all their bone mineral variables with age, whereas girls showed a decreasing trend from age 15 years. QUS had a significantly higher increase in standardized value with age than Ward's triangle BMD, but a significantly lower increase in standardized value with age than distal radius (cortical site) BMD. At other BMD sites we did not find any significant differences compared with QUS regarding changes with age. The measurements obtained by QUS, DXA and SXA, respectively, were divided into, quartiles. Of all subjects in the lowest quartile for QUS measurements, only 34 50% were also in the lowest quartiles for DXA and SXA measurements. In conclusion, QUS measurements of the calcaneus in children show similar results as for adult regarding the correlation with DXA and SXA; they also have a significant correlation with anthropometric data. QUS did not identify the same individuals with low bone mass as the X-ray techniques. PMID- 9850348 TI - Effect of inhaled corticosteroids on bone mineral density in childhood asthma: comparison of fluticasone propionate with beclomethasone dipropionate. AB - There is a dearth of data on long-term effects of inhaled corticosteroids (ICS) on bone architecture in childhood asthma. This study was designed to assess the possible effects of two different inhaled steroids on bone mineral density (BMD) in steroid-naive, prepubertal children. Twenty-three children were randomized to receive equipotent doses of either fluticasone propionate (100 micrograms twice daily) or beclomethasone dipropionate (200 micrograms twice daily). They were followed up over a period of 20 months with regular dual-energy X-ray absorptiometry scans for BMD. Densitometry of lumbar spine and total body showed a significant increase over time, which followed the normal patterns for growth. No difference was observed between the two subgroups. There was no change in fat distribution over time and no increase in percentage total body fat. As expected, girls had significantly higher total body fat. This absence of deleterious effects suggests that in the standard doses used neither beclomethasone nor fluticasone has any significant effect on bone density over a moderate period of time. Further studies should continue monitoring BMD through the critical years of bone mass accumulation during adolescence. PMID- 9850349 TI - How the new Hologic hip normal reference values affect the densitometric diagnosis of osteoporosis. AB - In February 1997, Hologic supplied new software to all QDR dual-energy X-ray absorptiometry (DXA) machines replacing the previous femoral normative reference database with the NHANES III normative data. In addition to changing the normative database (and therefore T-scores) for all regions of the hip, the new software has changed the primary region of interest from the femoral neck to the total hip. In the present study we examined how these changes influence the densitometric diagnosis of osteoporosis in a large clinical referral population (n = 2311, mean age 62.7 years). The patients had spine and hip DXA performed at either of two centers using a Hologic QDR-2000 over a 4-year period. T-scores were derived for each patient using both previous and current young normal reference databases. Intraindividual differences in T-scores were calculated. The prevalence of osteoporosis based on the two normative databases and the difference between the prevalence was calculated for each skeletal site. The average paired difference between current and previous T-scores at femoral neck is 0.64, the difference increasing with age. Using the new normative database, the percentage of osteoporotic patients decreases from 49% of all patients at the femoral neck to 28% at the femoral neck and 20% at the total hip. In conclusion, the densitometric diagnosis of osteoporosis will be affected in a significant proportion of women as a result of the implementation of the new hip normative database supplied by Hologic. Whether this will translate into fewer patients being treated remains to be seen. PMID- 9850350 TI - Effects of different regimens of sodium fluoride treatment for osteoporosis on the structure, remodeling and mineralization of bone. AB - We compared initial and final bone histomorphometric findings in 66 osteoporotic patients treated with sodium fluoride (NaF) according to three regimens, and in 7 osteoporotic patients who did not receive NaF. Fourteen patients received continuous NaF 75 mg/day (high-dose) with calcium 1500 mg/day for a mean of 41 months. Twenty-six patients received continuous NaF 50 mg/day (low-dose) with calcium 2000 mg/day for a mean of 15 months, either with (10 patients) or without (16 patients) vitamin D. Twenty-six patients received cyclical low-dose NaF, alternating with vitamin D, for a mean of 15 months and a total treatment duration of 28 months, of whom 14 were and 12 were not on NaF at the time of the second biopsy. Disregarding differences between regimens, there were significant increases in total and mineralized bone volume and trabecular thickness and nonsignificant decreases in these measurements in the control group. Fluoride induced bone formation was exclusively appositional with no evidence for the creation of new trabeculae. The effect of low-dose NaF on bone structure was the same when the same total dose was given continuously or intermittently, and when the patient was or was not taking vitamin D. The increases in total and mineralized bone volume but not trabecular thickness were greater with high-dose than with low-dose NaF. Low-dose NaF caused modest but significant increases in all osteoid indices, and modest but significant declines in adjusted apposition rate and osteoid maturation rate and no change in bone formation rate. With high dose NaF, the increase in BV/TV was greater but all indices of osteoid accumulation were much higher and all indices of impaired osteoblast function and mineralization were much lower, and 12 of 14 patients had some form of osteomalacia. This occurred also in 3 of 30 patients treated with low-dose NaF who were not taking vitamin D; the mean increase in osteoid thickness was significantly greater in these patients than in 22 low-dose patients who were taking vitamin D. We conclude: (1) The inconsistent effect of NaF in increasing bone strength is partly due to failure to restore connectivity in patients with severe bone loss and partly due to substantial osteoid accumulation. (2) Even low dose NaF causes impaired osteoblast function, but this is much greater with high dose prolonged therapy. (3) There is an unexplained discrepancy between the increase in bone formation implied by increases in spinal bone mineral and the lack of increase in bone formation measured histomorphometrically. (4) Defective mineralization is more closely related to the total cumulative dose of NaF than to the duration of treatment, and with low-dose treatment may be preventable by vitamin D. (5) Future clinical trials should be carried out with smaller doses of NaF and before there has been substantial loss of horizontal trabeculae in the spine. PMID- 9850351 TI - The Tromso Study: body height, body mass index and fractures. AB - Tall persons suffer more hip fractures than shorter persons, and high body mass index is associated with fewer hip and forearm fractures. We have studied the association between body height, body mass index and all non-vertebral fractures in a large, prospective, population-based study. The middle-aged population of Tromso, Norway, was invited to surveys in 1979/80, 1986/87 and 1994/95 (The Tromso Study). Of 16,676 invited to the first two surveys, 12,270 attended both times (74%). Height and weight were measured without shoes at the surveys, and all non-vertebral fractures in the period 1988-1995 were registered (922 persons with fractures) and verified by radiography. The risk of a low-energy fracture was found to be positively associated with increasing body height and with decreasing body mass index. Furthermore, men who had gained weight had a lower risk of hip fractures, and women who had gained weight had a lower risk of fractures in the lower extremities. High body height is thus a risk factor for fractures, and 1 in 4 low-energy fractures among women today might be ascribed to the increase in average stature since the turn of the century. Low body mass index is associated with a higher risk of fractures, but the association is probably too weak to have any clinical relevance in this age category. PMID- 9850352 TI - Volumetric bone density and geometry assessed by peripheral quantitative computed tomography in uremic patients on maintenance hemodialysis. AB - The aim of this study was to establish, by means of peripheral quantitative computed tomography (pQCT) at the distal radius, the existence of cortical and/or trabecular osteopenia, and to assess the integrity of bone geometry in uremic patients undergoing maintenance hemodialysis. Our results show a clearcut selective reduction in volumetric cortical density, more evident in women (p = 0.0001) than men (p = 0.030), which appears to be independent of age and menopausal status. Trabecular density was not significantly changed in either sex. Cortical density of the patients correlated inversely with age (p = 0.003), duration of dialysis (p = 0.002) and parathyroid hormone (PTH) levels (p = 0.03). Trabecular density correlated only with age. Normally, cortical density is age dependent and its reduction is accompanied by compensatory geometry changes. Compared with control subjects, in our female patients both cortical area and cortical thickness were reduced (p = 0.02 and 0.008), while cross-sectional area did not change (p = 0.67). Conversely, in the males only cross-sectional area was reduced (p = 0.02). In conclusion, in uremic patients we observed a selective cortical osteopenia, more evident in the female sex, and a sex-specific pattern of geometry impairment, with resultant apparent increased bone fragility in the uremic women. We suggest that the prolonged PTH excess could be responsible, directly and/or interacting with estrogen deficiency. PMID- 9850353 TI - The influence of physical activity and fractures on ultrasound parameters in elderly people. AB - In this cross-sectional study we investigated the relationship between ultrasound measurements in the calcaneus versus daily physical activity and fractures sustained in the past in elderly subjects. Ultrasound measurements were performed at both heels, which enabled us to examine determinants of differences between contralateral heels. Participants were 132 men and 578 women, aged 70 years and over (mean age and standard deviation (SD): 83 +/- 6 years), living in homes for the elderly (n = 343) or apartment houses for the elderly (n = 367). Broadband ultrasound attenuation (BUA) and speed of sound (SOS) were measured in the right and left calcaneus. The median difference (and interquartile range) between the two heels for BUA and SOS, expressed as a percentage of the mean value for each individual, was 9.6% (4.2-15.7%) and 1.0% (0.4-1.7%), corresponding to 25% and 40% of the study population SD, respectively. Greater differences in BUA between the two heels were associated with variables indicating poorer functional status, such as past fractures at the lower extremities. The level of daily physical activity was obtained by means of a questionnaire regarding household and leisure activities. Subsequently sumscores were calculated for daily physical activity and a subscore indicating weight-bearing physical activity. After adjustment for age, gender, residence, and body weight, physical activity scores were positively linearly related to both BUA and SOS. Each 5 point increase on the weight-bearing physical activity score, corresponding to, for example, walking for 2.5 h per week, was associated with a difference in BUA of 4.7 dB/MHz and in SOS of 5.1 m/s, which is similar to the differences associated with 10 kg higher body weight, or 10 years younger age. After the age of 50 years, 47 subjects had fractured a hip, 61 subjects had sustained another lower extremity fracture, 104 subjects a wrist fracture, and 62 subjects another upper extremity fracture. After adjusting for age, gender and residence, odds ratios for all fracture types in the lowest terciles of BUA and SOS versus the highest terciles ranged from 1.9 to 3.8. This study showed significant differences in ultrasound measurements between the left and the right heel, indicating that measurements at both sides are necessary for optimal evaluation of bone strength. Furthermore, after careful adjustments, ultrasound parameters had higher values with higher daily physical activity in elderly subjects and discriminated subjects with a history of fracture from those without. PMID- 9850354 TI - Hyperthyroidism influences ultrasound bone measurement on the Os calcis. AB - The objective of our study was to compare bone mineral density (BMD) measured by dual-energy X-ray absorptiometry (DXA) and quantitative ultrasound (QUS) parameters in women with hyperthyroidism and controls. In this cross-sectional study, QUS parameters and BMD values observed in untreated hyperthyroid patients were compared with data obtained from age-matched controls. Twenty-four women with Graves' disease were studied. Eight patients were postmenopausal. All patients had evidence of thyrotoxicosis as indicated by a raised total serum thyroxine and a suppressed serum thyroid stimulating hormone. BMD of the hip, lumbar spine and whole body, and body composition, were measured by DXA. Ultrasound evaluation on the os calcis was performed with an Achilles device. All measurements were performed before antithyroid therapy. The QUS parameters of BUA, SOS and Stiffness were significantly lower in hyperthyroid patients than in controls. Similar results were observed for the BMD of lumbar spine, femoral neck and total skeleton. Lean tissue and fat mass were also significantly decreased in hyperthyroid patients. In conclusion, these findings suggest that hyperthyroidism affects cortical and trabecular bone equally, as well as bone quality. QUS measurements may be helpful for assessing, using a simple and non-irradiating method, the bone effects of thyrotoxicosis. PMID- 9850355 TI - Comparative assessment of bone mineral measurements using dual X-ray absorptiometry and peripheral quantitative computed tomography. AB - A measurement of bone mass is the single most important determinant of future fracture. However, controversy exists as to which technique (dual X-ray absorptiometry (DXA) or peripheral quantitative computed tomography (pQCT), and which site of skeletal measurement (axial vs appendicular) provides the best prediction of fracture risk. The aims of this study were: (1) to determine the ability of pQCT to predict bone mass of the lumbar spine, proximal femur, and distal forearm measured using DXA, and (2) to compare the ability of DXA and pQCT to discriminate prevalent fractures in women with established osteoporosis. One hundred and sixty-five women were studied, including 47 with established osteoporosis (vertebral, hip or Colles' fractures) as well as 118 who had bone mass measurements to assess osteoporosis risk. Each subject had bone mass measured by DXA at the lumbar spine and femoral neck, and at the distal radius by both DXA and pQCT. In women with fractures, bone mass, when expressed as a standardized score, was in general lower using DXA compared with the appendicular skeleton measured using pQCT. Bone mass determinations at all sites were significantly correlated with each other. The highest correlation coefficients were observed within the axial skeleton. In women with fractures, the highest odds ratios were observed at skeletal regions measured using DXA. Likewise, the areas under the receiver-operating characteristic (ROC) curves were comparable at all skeletal regions measured using DXA; and were significantly greater than the areas under the ROC curves for pQCT measurements. In summary, the strongest discriminators of prevalent fractures were measurements using DXA. Measurements of bone mass at the appendicular skeleton, using either DXA or pQCT, were poorly associated with axial bone mass. PQCT has the poorer ability to discriminate persons with fractures, and appears to be less sensitive than measurements using DXA. PMID- 9850356 TI - Updated data on proximal femur bone mineral levels of US adults. AB - This paper describes data on bone mineral levels in the proximal femur of US adults based on the nationally representative sample examined during both phases of the third National Health and Nutrition Examination Survey (NHANES III, 1988 94), and updates data previously presented from phase 1 only. The data were collected from 14,646 men and women aged 20 years and older using dual-energy X ray absorptiometry, and included bone mineral density (BMD), bone mineral content (BMC) and area of bone scanned in four selected regions of interest (ROI) in the proximal femur: femur neck, trochanter, intertrochanter and total. These variables are provided separately by age and sex for non-Hispanic whites (NHW), non-Hispanic blacks (NHB) and Mexican Americans (MA). NHW in the southern United States had slightly lower BMD levels than NHW in other US regions, but these differences were not sufficiently large to prevent pooling of the data. The updated data provide valuable reference data on femur bone mineral levels of noninstitutionalized adults. The updated data on BMD for the total femur ROI of NHW have been selected as the reference database for femur standardization efforts by the International Committee on Standards in Bone Measurements. PMID- 9850357 TI - Improved delivery of inhaled steroids to the large and small airways. AB - The reformulation of asthma medications with non-ozone depleting propellants such as hydrofluoroalkane-134a (HFA) has provided the opportunity to apply new knowledge and inhaler technology to improve significantly the delivery of aerosol drugs to the respiratory tract. Beclomethasone dipropionate (BDP), the most commonly prescribed inhaled corticosteroid for asthma therapy, is effective therapy; however currently available chlorofluorocarbon (CFC)-BDP metered dose inhalers typically deliver no more than 10% of the inhaled drug to the lungs with the remainder deposited in the oropharynx. Compared with an average particle size of 3.5-4.0 microns for CFC-BDP, the new HFA-BDP formulation has an average particle size of 1.1 microns and a respirable fraction of approximately 60%. The lung deposition of 99mTc-radiolabelled HFA-BDP has been investigated in healthy volunteers and patients with asthma. Results showed that the HFA-BDP formulation reverses the pattern of distribution seen with CFC-BDP products, delivering most of the dose of inhaled steroid directly to the lungs rather than to the oropharynx and gut where it may lead to unwanted side-effects. As such, HFA-BDP is likely to achieve equivalent efficacy to existing CFC-BDP formulations with lower doses and with reduced potential for local adverse effects. PMID- 9850358 TI - Effect of changing the fine particle mass of inhaled beclomethasone dipropionate on intrapulmonary deposition and pharmacokinetics. AB - Reformulation of beclomethasone dipropionate (BDP) in the chlorofluorocarbon (CFC)-free propellant hydrofluoroalkane-134a (HFA) gave the opportunity to produce a solution formulation that provides a greater total mass of fine drug particles than the current CFC suspension metered dose inhaler (MDI). The HFA-BDP MDI was studied in three pharmacokinetic trials in asthmatic patients. Serum levels of BDP plus metabolites [total beclomethasone (total BOH) assay] were used to test whether the increased fine particle mass of HFA-BDP would result in improved intrapulmonary deposition and subsequent differences in serum profiles. Serum levels, maximum serum concentrations and area under the serum concentration time curves of total BOH following both single and multiple doses of HFA-BDP were similar to those obtained with approximately twice the dose of CFC-BDP. The observed lower bioavailability of CFC-BDP compared with HFA-BDP could be explained if most of each inhaled dose from the CFC-BDP MDI was swallowed and absorbed from the gastrointestinal tract, while most of each inhaled dose from the HFA-BDP MDI was absorbed from the lungs. Deposition studies have confirmed this explanation. These results suggest that asthmatic patients can be treated with lower total daily doses of drug from HFA-BDP extrafine aerosol than from CFC BDP. PMID- 9850359 TI - Efficacy of chlorofluorocarbon-free beclomethasone dipropionate 400 micrograms day-1 delivered as an extrafine aerosol in adults with moderate asthma. AB - Beclomethasone dipropionate (BDP) has been reformulated in a chlorofluorocarbon free propellant, hydrofluoroalkane-134a (HFA), resulting in an extrafine aerosol which gives improved drug delivery to the airways. The objective of this 6 week, placebo-controlled study was to evaluate the clinical efficacy of HFA-BDP 400 micrograms day-1 in 256 adult patients with moderate asthma. This is a lower daily dose than that recommended for existing BDP inhalers in current treatment guidelines for moderate asthma (NIH publication 95-3659). Another objective was to evaluate whether delivering the 400 micrograms dose as four actuations of 50 micrograms twice daily (HFA-BDP 50 micrograms) provided equivalent asthma control to delivering the dose as two actuations of 100 micrograms twice daily (HFA-BDP 100 micrograms) without the use of a spacer or holding chamber. Both active treatments produced a significant change from baseline in morning peak expiratory flow (PEF), which was significantly larger than that in the placebo group (P < or = 0.017) throughout the study. The mean changes from baseline in morning PEF at weeks 5-6 were 47.0 l min-1 in the combined HFA-BDP group and 16.5 l min-1 in the HFA-placebo group. The two dose strengths were statistically equivalent (P = 0.017 for equivalence testing). The active treatments also produced significant improvements compared with placebo in evening PEF, forced expiratory volume in the first second, forced expiratory flow over 25-75% of vital capacity, sleep disturbance scores and daily beta-agonist use. The study medication was well tolerated, with a low incidence of adverse events. The results from this study demonstrate that a daily dose of 400 micrograms HFA-BDP (given in 50 micrograms and 100 micrograms strengths) provides dose proportionality and effective control in patients with moderate asthma. PMID- 9850360 TI - Hydrofluoroalkane-134a beclomethasone dipropionate extrafine aerosol provides equivalent asthma control to chlorofluorocarbon beclomethasone dipropionate at approximately half the total daily dose. AB - The mandatory requirement to eliminate chlorofluorocarbons (CFCs) as propellants in pharmaceutical aerosols has provided the opportunity to enhance significantly the delivery of aerosol drugs to the respiratory tract. This randomized, parallel group, double-blind, double-dummy, multicentre study was undertaken to assess whether beclomethasone dipropionate (BDP) in hydrofluoroalkane-134a (HFA) provided equivalent control of moderately severe asthma to BDP in CFC but at approximately half the total daily dose, as might be expected from the improved lung deposition of the HFA-BDP extrafine aerosol. The novel study design included a 10-12 day run-in period to confirm that patients met established criteria of moderately severe asthma and were symptomatic on current therapy (inhaled beta agonist plus CFC-BDP 400-800 micrograms day-1). This run-in period was followed by a short course of oral steroid therapy (prednisolone 30 mg day-1 for 7-13 days) to demonstrate steroid responsiveness [> or = 15% improvement in morning peak expiratory flow (PEF)] and to provide a within-study baseline of improved asthma control. A total of 233 patients were randomized to treatment for 12 weeks with HFA-BDP 800 micrograms day-1 (116 patients) or CFC-BDP 1500 micrograms day-1 (117 patients). The mean change from oral steroid treatment in morning PEF with HFA-BDP was equivalent to that seen with CFC-BDP at all time intervals. Changes in other measures of pulmonary function, asthma symptom scores and beta-agonist use were equivalent in the two treatment groups throughout the 12 week treatment period. The safety profile of HFA-BDP compared favourably with that of CFC-BDP with no unexpected adverse events reported. Fewer patients on HFA-BDP than on CFC BDP had plasma cortisol levels below the normal reference range after 12 weeks of therapy (5.1% vs. 17.3%, respectively). In conclusion, HFA-BDP extrafine aerosol was found to provide equivalent control of moderately severe asthma to CFC-BDP at approximately half the daily dose with a favourable safety profile, suggesting an improved therapeutic ratio. PMID- 9850361 TI - Safety of hydrofluoroalkane-134a beclomethasone dipropionate extrafine aerosol. AB - Herein we assess the safety of an inhaled formulation of beclomethasone dipropionate (BDP) which uses the propellant hydrofluoroalkane-134a (HFA) for the treatment of asthma. Acute local tolerability (as assessed by the incidence of cough and mean forced expiratory volume after 1 s inhalation) was similar for both BDP and placebo formulated in either chlorofluorocarbon (CFC) or HFA propellants. A total of 43 patients were treated with HFA-BDP (0, 200, 400 or 800 micrograms day-1) or CFC-BDP (800 micrograms day-1) for 14 days and their 24 h urinary free cortisol (UFC) excretion and response to cosyntropin stimulation were measured. There was no difference in UFC between any of the doses of HFA-BDP and CFC-BDP. Adrenal responsiveness to cosyntropin stimulation was normal in all but one patient. Two large 12 week phase III trials compared HFA-placebo, HFA-BDP 400 micrograms day-1 and CFC-BDP 800 micrograms day-1 (n = 347), and HFA-BDP 800 micrograms day-1 and CFC-BDP 1500 micrograms day-1 (n = 233). For HFA-BDP at either dose, CFC-BDP 800 micrograms day-1 and HFA-placebo, the number of patients with morning plasma cortisol concentrations below normal was less than 4.4% but was 14.6% for CFC-BDP 1500 micrograms day-1. The incidence of adverse events was lower in the HFA-BDP groups than in the CFC-BDP groups (P = 0.012). The data indicate that, at doses of up to 800 micrograms day-1, HFA-BDP is at least as well tolerated as CFC-BDP. Other studies have found that equivalent efficacy is reached at lower doses of HFA-BDP than CFC-BDP. Equivalent efficacy at a lower dose and equivalent safety at the same dose imply that HFA-BDP may have a more favourable risk: benefit ratio than CFC-BDP when used at the recommended lower doses. PMID- 9850362 TI - Leukotriene antagonism in asthma and rhinitis. PMID- 9850363 TI - Total respiratory system compliance after thoracoplasty. AB - Recent attention has been directed towards the deleterious effects of the sequelae of pulmonary tuberculosis on respiratory function. Thus, a study was undertaken to find out to what extent the deformity of thorax after thoracoplasty increases the mechanical work of breathing. Compliance of the total respiratory system (Crs) was measured with a pressure-type body plethysmograph in nine post thoracoplasty tuberculosis patients, 15 patients with pulmonary tuberculosis and three patients with pleural diseases. Mean Crs in nine post-thoracoplasty tuberculosis patients was 0.0861 cmH2O-1, and that in six patients with moderately advanced pulmonary tuberculosis was 0.0841 cmH2O-1. Mean Crs (0.1301 cmH2O-1) in nine patients with a minimum lesion of tuberculosis was not different from the reference Crs values reported to date by several authors. Crs was significantly correlated with vital capacity, but not with functional residual capacity. There was no difference in specific Crs between post-thoracoplasty tuberculosis patients and patients with moderately advanced pulmonary tuberculosis. Crs of patients with a pleural disease was smaller than Crs of patients with a minimal lesion of pulmonary tuberculosis. The deformity of thorax after thoracoplasty caused a decrease of Crs. However, ventilation is not as severely impaired as anticipated from the degree of thorax deformity on chest X ray film, providing that movement of the diaphragm is unrestricted. PMID- 9850364 TI - Rapid diagnosis of smear-negative pulmonary tuberculosis via fibreoptic bronchoscopy: utility of polymerase chain reaction in bronchial aspirates as an adjunct to transbronchial biopsies. AB - Fibreoptic bronchoscopy was performed on 190 patients with chest radiographic lesions and negative sputum smears for acid-fast bacilli. Aside from obtaining transbronchial biopsies for histological examination, bronchial aspirate specimens were also tested for Mycobacterium tuberculosis complex DNA by a conventional polymerase chain reaction (PCR) technique. Of 177 transbronchial biopsies performed, a diagnosis was found in 64 cases [43 cases of tuberculosis (TB), 17 cases of lung carcinoma and four cases of other infective/inflammatory diseases] giving a diagnostic yield of 36.2%. PCR was positive in 105 of 108 finally diagnosed cases of TB and 22 of 82 non-TB cases. The sensitivity, specificity, positive predictive value and negative predictive value of PCR when applied to bronchial aspirate specimens for diagnosing smear-negative pulmonary TB were 97.2%, 73.2%, 82.7% and 95.2% respectively. Therefore, detection of M. tuberculosis complex DNA in bronchial aspirates by PCR might have an adjunctive place to transbronchial biopsies in the rapid diagnosis of smear-negative pulmonary tuberculosis. PMID- 9850365 TI - Prescription of nCPAP and nBIPAP in obstructive sleep apnoea syndrome: Italian experience in 105 subjects. A prospective two centre study. AB - Nasal continuous positive airway pressure (nCPAP) is the current treatment of obstructive sleep apnoea syndrome (OSAS). The indications of bilevel pressure support ventilation (BIPAP PSV) in OSAS patients remain controversial. The purpose of this investigation was to verify the frequency of prescription of BIPAP PSV in a group of OSAS patients when CPAP was ineffective or not tolerated during titration. The study included 286 consecutive patients > or = 18 years of age referred to two Sleep laboratories for sleep related breathing disorders (SRBD) between December 1994 and November 1995. Of these, 130 patients were enrolled and 105 (88 males, 77 females) with moderate to severe OSAS completed the study and were finally analysed. After a full night diagnostic polysomnography (PSGD), patients had a second full night PSG under nCPAP (PSGT). If nCPAP was not tolerated, or failed to correct breathing abnormalities during sleep, a second PSGT was performed, using a BIPAP PSV. Our study shows that nCPAP (mean 8.5 +/- 2.0 cmH20) was considered a satisfactory therapy in 81 patients (77%). Twenty four (23%) required BIPAP PSV (mean IPAP 13.9 +/- 2.9 cmH20). We found the highest prevalence of BIPAP in patients with OSAS associated to obesity hypoventilation syndrome (OHS) (11 of 17) and in OSAS associated to chronic obstructive pulmonary disease (COPD) (nine of 16). Patients treated with BIPAP PSV were more obese and had a higher PaCO2 and sleep-related desaturations and a lower FEV1, FVC, FEV1/FVC and PaO2. In conclusion our study shows that CPAP therapy in the effective therapeutic option in the majority of patients with OSAS. There is a subset of patients with OSAS associated to COPD or to OHS in whom BIPAP PSV may be a better treatment modality. PMID- 9850366 TI - Tension-time index of inspiratory muscles in COPD patients: role of airway obstruction. AB - Inspiratory muscle function has been shown to be related to general muscle weakness, weight loss, blood gas tensions, airway obstruction and hyperinflation. The aim of this study was to define (1) the factor that is the main determinant of the tension-time index of the inspiratory muscles (TTmus), and which this increases the risk of inspiratory muscle fatigue; and (2) whether a breathing strategy is adopted to avoid inspiratory muscle fatigue. Twenty-seven normal volunteers and 35 stable COPD outpatients (FEV1% predicted, range: 21-89%; and FRC/TLC, range: 49-77%) were studied. The TTmus was determined as follows: TTmus = PI/PImax.TI/Ttot, where Pi is the mean inspiratory pressure calculated from the mouth occlusion pressure (P0.1), PImax is the maximal inspiratory pressure, TI is the inspiratory time, and Ttot is the total time of the breathing cycle. COPD patients showed significantly lower PImax and higher P0.1, PI, PI/PImax, and TTmus than normal subjects. No patient had a TTmus value higher than the inspiratory muscle fatigue threshold of 0.33. The FEV1 was significantly correlated with TTmus and all its components in the patients. The FRC/TLC was also correlated with all components except PI. Body weight was only correlated with PImax. In a forward and backward stepwise regression analysis, FEV1 appeared to be the only significant factor explaining the variance of log (PI/PImax) and log (TTmus), whereas FRC/TLC was the principal determinant of PImax. In COPD patients, a non-linear relationship was found between TI and P0.1. A negative linear relationship was found between TI/Ttot and PI/PImax. In conclusion, although hyperinflation predominantly affected inspiratory muscle strength in a group of stable COPD patients with a wide range of severity, airway obstruction was the principal factor determining the magnitude of TTmus. In addition, in order to remain below the inspiratory muscle fatigue threshold, as the severity of airway obstruction increased, patients adopted a breathing strategy characterized by decreased TI/Ttot as inspiratory pressure demand increased. PMID- 9850367 TI - A 6-month comparison between formoterol and salmeterol in patients with reversible obstructive airways disease. AB - The aim of this randomized, open, parallel group study was to compare the clinical efficacy of formoterol dry powder capsule 12 micrograms b.i.d. and salmeterol dry powder 50 micrograms b.i.d. in the treatment of patients with reversible obstructive airways disease. The 6-month treatment was preceded by a 2 week run-in period. Morning pre-dose peak expiratory flow (PEF) during the last 7 days of treatment was the primary variable. Throughout the study, patients recorded morning and evening pre-dose PEF, use of rescue medication, respiratory symptoms and adverse events. Clinic visits were scheduled at monthly intervals. Of the 482 patients randomized (equal numbers in the two treatment groups), 428 completed the study. Four hundred and twenty-five patients were included in the efficacy analysis for the primary variable. For mean morning pre-dose PEF during the last 7 days of treatment, the 95% confidence interval (CI) for the treatment contrast formoterol minus salmeterol was included entirely in the pre-defined range of equivalence (CI limits = -8.69, +9.841 min-1). This was also the case for the morning PEF during the last week before each clinic visit. For mean evening pre-dose PEF, the estimated treatment contrasts showed a trend towards superiority of formoterol over salmeterol, which became statistically significant at 2, 3 and 4 months (P < 0.05; estimated contrasts 7.27, 10.45 and 10.511 min-1, respectively). No treatment group differences were found in use of rescue medication and respiratory symptom scores. The incidence of adverse events was similar in the two groups. These findings demonstrate that formoterol 12 micrograms b.i.d. and salmeterol 50 micrograms b.i.d., both formulated as dry powders, have similar long-term efficacy and safety profiles in patients with reversible obstructive airways disease. PMID- 9850368 TI - Comparison of the severity of sleep-disordered breathing in Asian and Caucasian patients seen at a sleep disorders center. AB - Race can be considered a risk factor for sleep-disordered breathing (SDB), with higher prevalences and greater severity of the disorder documented among persons of certain racial groups compared with others. Based on clinical observation, it was hypothesized that, other risk factors being equal, Asian patients with SDB have greater severity of their illness compared to Caucasian patients. A cross sectional study was conducted at a sleep disorders clinic involving 105 Asian patients diagnosed as having SDB after undergoing polysomnography and 99 similarly diagnosed Caucasian patients matched for the following variables: age, gender and body mass index (BMI). The main outcome measure of interest was objective assessment of severity based on polysomnographic data of respiratory disturbance index (RDI) and minimum oxygen saturation (SaO2) during sleep. Symptom scores between patients of the two racial groups were also compared. There were significantly larger proportions of Asians compared to Caucasians with severe obstructive sleep apnea (OSAS) as defined by respiratory disturbance index (RDI) > or = 50 (25.0% vs 11.1%; P = 0.0288) or minimum oxygen saturation (SaO2) < or = 69% (20.6% vs 4.2%; P = 0.0113). The mean minimum SaO2 was significantly lower (P = 0.0001) while the mean (log transformed) esophageal pressure (Pes) value was significantly higher (P = 0.0090) in the Asian group. Logistic regression analysis showed that race was associated with severe SDB (RDI > or = 50) independent of age, sex and BMI. The estimated odds ratio for Asians having severe OSAS compared with Caucasians was 2.51 [95% Confidence Interval (CI) 0.98 6.64]. There was no significant difference in the severity of questionnaire-based symptoms of snoring, apneas during sleep and the median Epworth scores between Asian and Caucasian patients. Based on objective polysomnographic results, Asian patients with OSAS have greater severity of their illness compared to Caucasian patients matched for age, gender and BMI. There was, however, no significant difference in severity of questionnaire-based symptoms between Asian and Caucasian patients with SDB. PMID- 9850369 TI - Symptoms and pulmonary function in asthma. AB - The relationship between symptoms and pulmonary function in asthma is important if the latter is to be held relevant to management guidelines and their audit. Associations between reported symptoms, pulmonary function and therapy were studied in 824 asthmatics (mean FEV1 75.4% predicted; best FEV1 84.6% predicted; and actual/best peak flow (PEF) 87.5%). Bronchodilator usage (reflecting symptomatic wheeze) was evenly distributed up to eight times daily; 22.5% of subjects had nocturnal disturbance and 46.3% persistent daytime symptoms. The univariate relationships between symptoms and function were generally closer with best rather than actual/best. They were further explored using quintiles of function. Symptoms were consistently less as best function increased, but were highly significantly greater in the fifth than in the third and fourth quintiles of actual/best FEV1. There was a trend to a similar U-shaped relationship of actual/best PEF with nocturnal disturbance and daytime symptoms. Best function is a good determinant of expected symptom load in an asthmatic population. Below 85% actual/best function reflects the prevalence of symptoms. In asymptomatic patients a level of at least 85-90% is a useful check of physiological control but will not exclude some symptomatic patients, irrespective of best function. PMID- 9850370 TI - Effects of positive end-expiratory pressure (PEEP) in cardiac surgery patients. AB - The role of positive end-expiratory pressure (PEEP) in the postoperative course of cardiac surgery patients remains questionable. In this prospective study, we examined the effect of different levels of PEEP on arterial oxygenation, SvO2 and PvO2 values, and on haemodynamic indices, during the early postoperative period in cardiac surgery patients. Upon transfer to the ICU, 67 adult patients with normal preoperative respiratory status were randomly assigned to receive zero PEEP (Group A), 5 cmH2O (Group B), or 10 cmH2O PEEP (Group C) during mechanical ventilatory support. PaO2/FIO2 ratio, mixed venous PvO2 and SvO2, and cardiac index, were measured 30 min, 4 h and 8 h after application of mechanical ventilation in the ICU, just prior to extubation, half an hour after extubation, and 4 h post-extubation. We found no statistically significant differences (P = n.s.) in arterial oxygenation expressed by PaO2/FIO2 ratio, SvO2 and PvO2 values, and in cardiac index among the three groups at any study interval. We conclude that low levels of PEEP have no advantage over zero PEEP in improving gas exchange in the early postoperative course of patients following open heart surgery. PMID- 9850371 TI - Neutrophils infiltrating bronchial epithelium in chronic obstructive pulmonary disease. AB - In order to characterize neutrophil and eosinophil presence in the airways of patients with chronic obstructive pulmonary disease (COPD), bronchoscopy with bronchial washings and bronchial biopsies was performed in 12 smoking stable COPD subjects and 18 normal non-smoking control subjects. Bronchial biopsies were examined by light microscopy using plastic embedding and histochemical techniques to identify different cell types. Bronchial washing fluid of COPD patients was characterized by a predominance of neutrophils (P = 0.001), and a slight, but significant (P = 0.03), increase of eosinophil fraction. Subjects with COPD had higher number of neutrophils in the epithelium (P = 0.01), and eosinophils in the lamina propria (P = 0.01) than did control subjects. The thickness of reticular basement membrane was increased for COPD patients in comparison to control subjects (P = 0.01). The present study provides evidence of neutrophil infiltration both in bronchial washing and bronchial epithelium of patients with COPD, suggesting that the source of neutrophils in airway lumen may be the bronchial mucosa. Although less common than in asthma, airways of COPD subjects reveal eosinophil presence and airway remodelling. PMID- 9850372 TI - Diabetic retinopathy in adult patients with cystic fibrosis-related diabetes. PMID- 9850373 TI - Multiple drug-resistant Nocardia asteroides isolated from a patient with pulmonary nocardiosis. PMID- 9850374 TI - Chest CT scanning: utility in lung endometriosis. PMID- 9850375 TI - Solitary columnar bronchial papilloma: a rare endoscopic finding. PMID- 9850376 TI - Thymic carcinoid with mucinous stroma: a case report. PMID- 9850377 TI - Primary mediastinal nonseminomatous germ cell tumour in an adult female. PMID- 9850378 TI - Idiopathic bronchiolitis obliterans with organizing pneumonia presenting with adult respiratory distress syndrome. PMID- 9850379 TI - Diffuse pulmonary ossification and recurrent spontaneous pneumothorax in a patient with bronchial asthma. PMID- 9850380 TI - Pleomorphic adenoma and severe tracheal obstruction. PMID- 9850381 TI - Appearance of central apnoea in a patient treated by auto-CPAP for obstructive sleep apnoea. PMID- 9850382 TI - Royal Society of Tropical Medicine and Hygiene meeting at Manson House, London, 19 February 1998. Amoebic disease. Entamoeba dispar, an organism reborn. AB - Entamoeba dispar was described by Emile Brumpt in 1925, and was promptly dismissed as a synonym of E. histolytica by almost all his contemporaries. In the 1970s, however, evidence began to accumulate indicating that Brumpt may have been correct and E. dispar is now accepted as a distinct species. This review introduces the organism's history and briefly summarizes what we currently know about its biology. The acceptance of E. dispar has profound implications for our understanding of the epidemiology of amoebiasis and, ultimately, may give us an insight into what makes E. histolytica a pathogen. PMID- 9850384 TI - Long-lasting anti-mosquito efficacy of a commercially impregnated bed net. PMID- 9850383 TI - Permethrin-treated bed nets do not have a 'mass-killing effect' on village populations of Anopheles gambiae s.l. in The Gambia. AB - In The Gambia, the use of permethrin-treated bed nets has led to a reduction in morbidity and mortality from malaria in children. However, no clear evidence has been found for a 'mass-killing effect' on the mosquito vectors as a result of this intervention. Two further entomological studies to investigate this phenomenon have been carried out. In one study, 20 villages were paired so that bed nets in one member of each pair were treated with permethrin. In the other, a cross-over design was used in which treated and untreated bed nets were exchanged between 2 villages. Longevity, biting rate and resting density of the malaria vector population and sporozoite rates were assessed in both studies. Malaria vectors were equally abundant and long-lived, and as likely to be infective, in villages with treated bed nets as in those with untreated nets. However, a clear reduction in the density of the indoor-resting population of mosquitoes in rooms with treated bed nets was found, probably reflecting the excito-repellency of the insecticide. This study confirmed that, in The Gambia, the protection against death and morbidity from malaria seen in children using treated bed nets must be due primarily to personal protection rather than to a 'mass-killing effect' on the mosquito vector population at a village level. PMID- 9850385 TI - Environmental and entomological risk factors for the development of clinical malaria among children on the Kenyan coast. AB - Several malariometric studies have examined the impact on human-vector contact of house construction, demographics, bed net and insect repellent use. However, few studies have documented the significance of these proximate determinants on the risks of clinical disease. We undertook a matched case-control study of the risks of both mild clinical malaria and severe life-threatening malaria according to a range of putative factors which would influence the frequency of child-vector encounters in Kilifi district on the Kenyan coast. Among 394 severe disease cases, 380 age-matched mild disease cases, and their respective location and age matched community controls, we were unable to demonstrate any statistically significant effect upon disease outcome of house construction, presence of domestic animals, or bed net use. Higher population density within a 250 m radius of the homes conferred significant protection from the risks of developing severe malaria compared to community controls. The risks of developing severe malaria compared to the community controls and the transition from mild to severe disease were statistically significantly lower in those who reported use of mosquito coils, local repellents or aerosol insecticides. We concluded that it is likely that the impact of household features on disease outcome is dependent upon both the density of infecting mosquitoes and acquired immunity within a given locality. PMID- 9850386 TI - Predicting the impact of school-based treatment for urinary schistosomiasis given by the Ghana Partnership for Child Development. AB - Mathematical models can be used to predict the impact of interventions to control infectious diseases. In this paper, an epidemiological model is used to predict the impact of chemotherapy of school-age children infected with Schistosoma haematobium, in a programme conducted by the Ghana Partnership for Child Development in the Volta Region, Ghana. Existing data were used to validate the predictions of the model, demonstrating convincingly the ability of the model to make correct predictions. Predictions of trends in mean egg count, infection prevalence and prevalence of heavy infection (> 50 eggs/10 mL urine) were then made for the period 1997-1999, and will be compared to the data collected in the programme in the future. PMID- 9850387 TI - The relationship between infection and disease in Wuchereria bancrofti infection in Ghana. AB - The relationship between infection and clinical disease in Wuchereria bancrofti infection was investigated in a community-based study in different endemic areas in Ghana. At the individual level, there was no association between acute adenolymphangitis and infection (microfilaraemia) status. There was a negative association between infection status and lymphoedema/elephantiasis, but a positive association with hydrocele; however, the intensity of infection was negatively associated with both elephantiasis and hydrocele. The community prevalence of infection was strongly associated with the prevalence of clinical filariasis (especially hydrocele). There was a strong positive association between the prevalence of infection in males and the odds of a case of hydrocele being microfilaraemic, suggesting that there is no acquired immunity to reinfection in cases of hydrocele. The pathophysiologies of elephantiasis and hydrocele may therefore differ from one another, and require further investigation. PMID- 9850388 TI - Seroprevalence of Rickettsia in a gold-panning population in north-eastern Gabon. PMID- 9850389 TI - Detection of Plasmodium falciparum gametocytes with the OBC test and Giemsa stained thick blood films for malaria transmission studies in Cameroon. PMID- 9850390 TI - Use of the polymerase chain reaction to assess the success of visceral leishmaniasis treatment. AB - We investigated whether the polymerase chain reaction (PCR) performed with aspirates of bone marrow or lymph node can be used as a test of cure of visceral leishmaniasis (VL). Sixty-one VL patients who had received supervised treatment with sodium stibogluconate in the health centre of Medecins sans Frontieres (MSF) Holland in Um-Kuraa, eastern Sudan, were studied. Immediately after treatment, no parasite could be demonstrated by microscopy in aspirates of bone marrow or lymph node. In contrast, PCR detected Leishmania deoxyribonucleic acid in 50 of the 61 lymph node aspirates (82%). Forty-nine patients were examined 3 and 6 months later; the other 12 were reported to be alive but had left the area. With 10 of these 49 patients, the PCR was negative and the patients remained free from signs and symptoms of VL; they were apparently cured. Of the 39 patients with a positive PCR after treatment, 14 (36%) developed post kala-azar dermal leishmaniasis and 9 (23%) had a recurrence of VL symptoms with reappearance of parasites in the aspirates. Four relapsed patients subsequently died of VL. We concluded that the PCR on lymph node aspirates can be used to assess treatment and cure of VL. The fact that 23 of 49 patients who received standard supervised treatment were not completely cured indicated that there is a need to investigate extended or alternative treatments. PMID- 9850391 TI - Evaluation of the Eiken latex agglutination test for anti-Toxoplasma antibodies and seroprevalence of Toxoplasma infection among factory workers in Addis Ababa, Ethiopia. AB - Sera from 170 factory workers aged 18-45 years enrolled in a pilot study of human immunodeficiency virus 1 (HIV-1) infection in Addis Ababa, Ethiopia, were screened for anti-Toxoplasma immunoglobulin G antibodies by the Sabin-Feldman test (reference standard) and the Eiken latex agglutination test (under evaluation for use in developing countries). Based on the Sabin-Feldman test, the prevalence of anti-Toxoplasma antibodies was 80.0% (95% confidence interval 73.9 86.1%). The sensitivity and specificity of the Eiken latex agglutination test were 96.3% and 97.1%, respectively, showing its validity for the detection of anti-Toxoplasma antibodies. The prevalence of antibodies did not differ between individuals infected and uninfected with HIV-1 (74.2% versus 83.3%, P > 0.05). However, antibody titres were higher in HIV-infected persons than in those who were uninfected (P < 0.001). Based on these findings, we expect that toxoplasmic encephalitis will be a common opportunistic infection among HIV-infected Ethiopians, and chemoprophylaxis with co-trimoxazole may be beneficial to those with low CD4+ T cell counts. The prognostic significance of high titres of anti Toxoplasma antibodies remains to be established among Ethiopian HIV-infected individuals. PMID- 9850392 TI - PCR-ELISA for the detection of Brugia malayi infection using finger-prick blood. AB - A polymerase chain reaction assay based on the enzyme-linked immunosorbent assay (PCR-ELISA) has been developed to detect Brugia malayi infection in an area of low endemicity in Malaysia. Blood samples from 239 subjects were tested: 192 amicrofilaraemic individuals, 14 microfilaraemic persons and 3 chronic elephantiasis cases from endemic areas and 30 city-dwellers (non-endemic controls). PCR products were examined by ELISA and Southern hybridization. In the PCR-ELISA, digoxigenin-labelled PCR products were hybridized to a biotin-labelled probe. This was followed by incubation in streptavidin-coated microtitre wells and detection using anti-digoxigenin-peroxidase and ABTS [2,2'-azinobis(3 ethylbenzthiazoline-6-sulphonic acid)]. All microfilaraemic samples were positive by PCR-ELISA and Southern hybridization and all samples from non-endemic subjects and chronic elephantiasis patients were negative. The PCR-ELISA detected 12 times as many B. malayi infections as did thick blood film examination. Nineteen of the 194 samples from the endemic area gave positive results by both PCR-ELISA and Southern hybridization, and an additional 5 samples were positive by PCR-ELISA only. The PCR-ELISA was specific and sensitive, detected more infections, and was more reproducible than Southern hybridization. PMID- 9850393 TI - Evaluation of the filter paper blood collection method for detecting Og4C3 circulating antigen in bancroftian filariasis. AB - Serological diagnosis of filariasis is generally known to be more reliable than detection of microfilariae. The recently developed Og4C3 enzyme-linked immunosorbent assay (ELISA) for detecting Wuchereria bancrofti circulating antigen has been shown to be very sensitive in diagnosing filiariasis using serum samples. The commercially available form of this ELISA, using whole blood collected on filter paper, has not been validated independently. We evaluated the sensitivity of this new method against standard 20 microL night blood films in 1808 paired samples from 18 communities in different endemic areas of Ghana. The diagnostic performance of the method was consistently low in all but 2 communities (sensitivity = 50.3%). This method of diagnosing filariasis is not suitable for field use in its present form. PMID- 9850394 TI - A specific antigen-detection ELISA for the diagnosis of human neurocysticercosis. The Cysticercosis Working Group in Peru. AB - An enzyme-linked immunosorbent assay (ELISA) for the detection of antigen secreted by viable Taenia solium metacestodes (Ag-ELISA) was applied to 43 pre treatment and 47 follow-up cerebrospinal fluid (CSF) samples from Peruvian patients with neurocysticercosis demonstrated by computed tomography and enzyme linked immunoelectrotransfer blot assay. The sensitivity of the assay was 86%. Negative pre-treatment results in the Ag-ELISA test were restricted to patients with only a single live cyst or only enhancing lesions. Patients with hydrocephalus had higher levels of circulating antigen. There was no difference between antigen levels in CSF taken before and immediately after treatment (day 14). Levels of parasite antigen were significantly positively correlated with the number of live cysts detected by tomography and were also proportional to the number and intensity of antibody reactions recognized by the immunoblot diagnostic test. In contrast, there was a negative correlation with the number of enhancing lesions revealed by tomography, supporting the hypothesis that enhancing lesions correspond to a terminal, moribund stage of the parasite. The use of antigen-detection tests specific for viable metacestodes has immediate utility in the clinical context, not only providing important information on the viability of the parasites but also leading to an improved understanding of the pathogenesis of neurocysticercosis before and after drug treatment. PMID- 9850395 TI - 'Improved sensitivity of direct microscopy for detection of acid-fast bacilli in sputum in developing countries. AB - Diagnosis of pulmonary tuberculosis depends on the bacteriological examination of sputum. Sputum smear microscopy is efficient and can confirm the disease. However, direct microscopy of sputum, though rapid, has low sensitivity. There is, therefore, an urgent need to develop rapid, much more sensitive and specific methods. In a field study we collected sputum samples from 488 tuberculosis suspects and compared the results of examining smears prepared after treatment with sodium hypochlorite (NaOCl) and concentration of bacteria by centrifugation and direct staining with Ziehl-Neelsen stain. Direct smears stained with auramine phenol were the reference standard. The use of NaOCl increased the sensitivity from 43.4% to 76.3%, with a specificity of 100% for both methods. The method is simple and cheap. As a potent disinfectant, NaOCl also reduces the risk of laboratory-acquired infection. Its application would increase the efficiency of tuberculosis control programmes. PMID- 9850396 TI - The ratio of reactive nitrogen intermediates to tumour necrosis factor and clinical outcome of falciparum malaria disease. AB - Serum levels of reactive nitrogen intermediates (RNI; nitrate + nitrite), interferon gamma (IFN gamma) and tumour necrosis factor (TNF) were measured in 177 Papua New Guinean children with different clinical manifestations of malaria. The groups investigated were asymptomatic parasitaemic, mild malaria, cerebral malaria survivors and cerebral malaria non-survivors. The levels of TNF were highest among the cases of cerebral malaria who died and lowest among the asymptomatic parasitaemic children (mean log TNF levels 2.183 pg/mL vs. 1.455 pg/mL; P = 0.001). Similarly, the levels of IFN gamma were highest among the cerebral and lowest among the asymptomatic patients (mean log TNF levels 0.338 pg/mL vs 0.054 pg/mL; P < 0.0001). RNI levels were high among both the asymptomatic parasitaemic group and those who died due to cerebral malaria (mean log RNI levels 1.56 microM vs. 1.412 microM; P = 0.18). The ratio of RNI to TNF, however, was significantly higher among the asymptomatic parasitaemic children and lowest among those who died due to cerebral malaria (mean log (RNI:TNF) ratio 0.118 vs. -0.789; P < 0.001). We concluded that the ratio of serum RNI to serum TNF is a more useful indicator of outcome of falciparum malaria in this population than the absolute levels of either alone. PMID- 9850397 TI - Pulmonary oedema in vivax malaria. PMID- 9850398 TI - Microsporidiosis in HIV-seronegative patients in Mali. PMID- 9850399 TI - Primary HIV infection diagnosed in South Africa masquerading as another tropical disease. AB - Eleven patients referred to a hospital in South Africa with suspected tropical diseases such as malaria, typhoid fever and South African tick bite fever were found to be suffering from primary human immunodeficiency virus (HIV) infection. Hospital records were reviewed retrospectively in those acutely ill, febrile patients where a clinical suspicion of HIV seroconversion existed and no other diagnosis could be found. A history of recent travel, particularly to malarious areas, was given by most of these patients. The clinical presentation was dominated by high fevers and headaches. The most helpful pointers to primary HIV infection included a characteristic palatal enanthem, leucopenia and thrombocytopenia. Ironically, the history of recent travel appeared to have confounded the diagnosis despite the fact that travel has often been associated with the acquisition of HIV in Africa. Recognition of primary HIV infection masquerading as a tropical disease may result in more frequent diagnosis of this serious condition. PMID- 9850400 TI - Cutaneous leucocytoclastic vasculitis from contact with Hylesia moths (Lepidoptera: Saturniidae). PMID- 9850401 TI - Artemisinin derivatives in the treatment of falciparum malaria in pregnancy. AB - An artemisinin derivative (artesunate or artemether) was used for the treatment of multidrug-resistant Plasmodium falciparum malaria in 83 Karen pregnant women in Thailand; 55 women were treated for recrudescent infection following quinine or mefloquine, 12 for uncomplicated hyperparasitaemic episodes, and 16 had not declared their pregnancy when treated. The women were followed weekly until delivery. Artesunate and artemether were well tolerated and there was no drug related adverse effect. Recrudescence within 42 d occurred in 16% of the treated episodes. Overall 73 pregnancies (88%) resulted in live births, 3 (4%) in abortions and 2 (3%) in still births, and 5 women were lost to follow-up before delivery. There was no congenital abnormality in any of the newborn children, and the 46 children followed for more than one year all developed normally. PMID- 9850402 TI - The pharmacokinetics of artemisinin suppositories in Vietnamese patients with malaria. AB - Eight male Vietnamese malaria patients received 600 mg of artemisinin in a single dose of 3 suppositories containing 200 mg each; 24 h later they received a single oral dose of mefloquine, 15 mg/kg. Plasma artemisinin concentrations were measured until 24 h after dosing, and parasites were counted until none could be detected. Artemisinin concentration versus time curves of all subjects were analysed with model-independent methods. Mean Cmax was 108 micrograms/L (SD = 60, range 29-169), mean tlag was 0.3 h (SD = 0), mean tmax was 6.5 h (SD = 3.9, range 2-14). By comparing the area under the concentration-time curve with that found in a previous study on oral artemisinin, average bioavailability relative to oral administration was estimated to be approximately 30%. Median parasite clearance time was 24 h (range 24-72). We concluded that therapeutic blood concentrations of artemisinin can be reached after rectal administration. There was a large inter-individual variation in blood concentrations attained. The dose given by rectal administration should probably be twice the usual oral dose, i.e., at least 20 mg/kg of body weight twice daily. PMID- 9850403 TI - An open randomized clinical study of intrarectal versus infused Quinimax for the treatment of childhood cerebral malaria in Niger. AB - The intrarectal route has been shown to be an alternative to parenteral therapy for the treatment of acute uncomplicated malaria. We conducted an open randomized clinical study of intrarectal Quinimax (a Cinchona alkaloids association) (20 mg/kg, then 15 mg/kg every 8 h) vs. intravenous Quinimax (8 mg/ kg infused over 4 h every 8 h) for 2 d in 76 children (39 in the intrarectal and 37 in the infusion groups) with cerebral falciparum malaria in Niger. This treatment was followed by oral chloroquine (10 mg/kg/d for 3 d). The primary end points of the study were fatal outcome and coma recovery time. In the intrarectal group, 35 children were cured (90%) and 4 died. In the infused group, 28 were cured (76%) and 9 died; mean coma recovery times were 34.6 h (SD = 12.8) and 33.0 h (SD = 14.1) for the intrarectal and infused groups, respectively. None of the differences was significant. Both treatments were well tolerated and no anal irritation was observed with intrarectal Quinimax. These findings suggest that intrarectal Quinimax can be an alternative to intravenous administration for rapid onset childhood cerebral malaria in the rural tropics, where the safety of parenteral administration cannot be guaranteed. PMID- 9850404 TI - Comparative efficacy of chloroquine plus chlorpheniramine and halofantrine in acute uncomplicated falciparum malaria in Nigerian children. AB - In the face of growing chloroquine resistance of Plasmodium falciparum, efforts to prolong the clinical usefulness of the drug have partly concentrated on its combination with potential resistance-reversing compounds. However, clinical studies on such combinations have been limited. We have compared the efficacy of halofantrine, an arylaminoalcohol effective in chloroquine resistant malaria, and a combination of chloroquine plus chlorpheniramine, a histamine H1 receptor antagonist which reverses chloroquine resistance of P. falciparum in vitro and in vivo, in 100 children with acute symptomatic uncomplicated falciparum malaria in an area in Nigeria where the rate of chloroquine resistance is 35-45%. Both chloroquine plus chlorpheniramine and halofantrine produced similar parasite and fever clearance times and cure rates (96%). Both treatment regimens were relatively well tolerated. Pruritus was commoner in patients treated with chloroquine plus chlorpheniramine than in those treated with halofantrine. Intravascular haemolysis occurred in one patient, and abdominal pain with or without diarrhoea occurred in 4 patients, treated with halofantrine. In vitro, the chloroquine resistance of P. falciparum isolates obtained from the patients was reversed by verapamil. All patients with isolates which were chloroquine resistant in vitro were cured by either therapy. These results indicate that chloroquine plus chlorpheniramine is as effective as halofantrine and is without overt deleterious effect in treating acute uncomplicated chloroquine-resistant falciparum malaria in children, and may be a clinically useful alternative for this purpose in Nigeria. PMID- 9850405 TI - Cardiac effects of halofantrine in children suffering from acute uncomplicated falciparum malaria. AB - The cardiac effects of halofantrine were assessed in 42 children with acute symptomatic uncomplicated Plasmodium falciparum malaria by electrocardiographic (ECG) and clinical monitoring over a period of 14 d. The children were treated with oral halofantrine 8 mg/kg body weight every 6 h for 3 doses. There was significant prolongation of the P-R interval (compared with the pre-treatment value) only at 8 h after drug administration. However, first degree auriculoventricular (AV) block occurred in 2 children at 8 h or 8 and 48 h, and second degree AV block in another child at 48 h. There was significant prolongation of the Q-Tc interval at 8, 16, 24, 48 and 72 h after treatment; the proportions of children with Q-Tc interval > 0.44 s were also significantly higher at all these times except 72 h. Rhythm disturbance was rare. There was no significant ECG change at 168 or 336 h. Despite the ECG abnormalities, there was no clinical symptom. These findings indicate that, in children, the currently recommended dose of halofantrine for the treatment of falciparum malaria may produce serious cardiac side effects. PMID- 9850406 TI - Treatment of late stage rhodesiense trypanosomiasis using suramin and eflornithine: report of six cases. PMID- 9850407 TI - Reversibility of Schistosoma mansoni-associated morbidity after yearly mass praziquantel therapy: ultrasonographic assessment. AB - A parasitological, clinical and ultrasonographic longitudinal study was undertaken in 1993 in a focus hyperendemic for Schistosoma mansoni infection in the central highlands of Madagascar. All the inhabitants were systematically treated with praziquantel. A complete examination and treatment were repeated each year. Among the 289 villagers who underwent the complete 3 years' follow up, 65.9% excreted eggs at the initial survey and the mean egg count of infected individuals was 202 eggs/g. In 1996, the prevalence of infection was 19.3% with a mean egg count of 27 eggs/g and, among inhabitants aged > 44 years, only one was found to be infected. The proportion of individuals complaining of bloody stool decreased from 24.9% in 1993 to 8.4% in 1996. Compared to the initial clinical examination, the age-adjusted prevalence of splenomegaly was significantly lower in 1996, but remained high: 62% in the 10-14 years age group and 59% in individuals aged > 24 years. Ultrasonographic examination after 3 years of praziquantel therapy showed a marked decrease of the overall prevalence of schistosomal hepatic fibrosis, from 28% in 1993 to 10.3% in 1996. This improvement had already been achieved during the second year of follow-up for most subjects. Usually, the reversal of morbidity affected individuals classified as stage 1 at the beginning of the study. Stage 3 was not observed in the last 2 surveys. One patient's ascites disappeared during the follow-up, associated with a significant reversal of periportal fibrosis. Our results indicate that repeated praziquantel therapy can lead to improvement of liver morbidity and the prevention of the development of schistosomal hepatic fibrosis, even in an old established hyperendemic focus. PMID- 9850408 TI - Impact of repeated large scale ivermectin treatments on the transmission of Loa loa. AB - We have studied the impact of large-scale treatment with ivermectin on the transmission of loiasis in a forest village in south Cameroon where loiasis was highly endemic, with a prevalence of 30%. After one year of parasitological and entomological surveillance without treatment, all consenting residents aged > 5 years received ivermectin 200 micrograms/kg every 3 months. For ethical reasons, treatment was interrupted after 2 years, but parasitological and entomological surveillance continued for 18 months after the end of treatment. The prevalence of loiasis was reduced to < 10% and the mean microfilaraemia decreased by 90% in 2 years. The prevalence and average intensity of infection remained stable during the 18 months after treatment ended. Two vector species were identified, Chrysops dimidiata (representing about 90% of the fly population) and C. silacea. The infection rate (all stages) in Chrysops decreased by 75% and the infective rate (percentage of Chrysops harbouring third-stage larvae of Loa loa in the head) decreased by 85% in C. dimidiata and became zero in C. silacea. After the end of treatment, the infection and infective rates increased gradually. Large-scale treatment seemed an efficient method for the control of L. loa transmission provided high drug coverage was achieved. Nevertheless, because of the high risk of adverse effects when using the current microfilaricidal drugs, such a strategy remains unacceptable. PMID- 9850409 TI - Decrease in Mansonella perstans microfilaraemia after albendazole treatment. PMID- 9850410 TI - A randomized clinical trial to compare the efficacy of erythromycin, ampicillin and tetracycline for the treatment of cholera in children. AB - To compare the clinical outcome of treatment of cholera in children with ampicillin, erythromycin or tetracycline, a double-'blind' randomized four-cell trial was carried out in Bangladesh. Ampicillin was chosen as additional therapy for acute respiratory tract infection, present in many subjects with diarrhoea. One hundred and eighty-four children aged 1-5 years who were not wasted, with diarrhoea of duration < 48 h, signs of some or severe dehydration, dark-field stool microscopy demonstrating Vibrio cholerae, and a baseline purging rate > 4 mL/kg/h over 6 h were enrolled in the study. Ampicillin, tetracycline, erythromycin or placebo were given orally every 6 h for 3 d. After 3 d of antibiotic treatment, diarrhoeal stool volume was significantly reduced in all antibiotic groups, with mean volumes per kg body weight as follows: tetracycline, 318 mL (SEM = 50), ampicillin, 335 mL (SEM = 30); erythromycin, 323 mL (SEM = 25); placebo, 498 mL (SEM = 37). Compared to tetracycline, the clinical recovery rates by 96 h were 75% with placebo, 91.3% with ampicillin, and 95.7% with eythromycin. Compared to tetracycline, the total mean times to recovery were increased by 66% with placebo (P < 0.001), 25% with ampicillin (P < 0.017), and 9% with erythromycin (P = 0.37). These results indicated comparable clinical efficacy of tetracycline, ampicillin and erythromycin. We therefore recommend that, unless V. cholerae is resistant, ampicillin should be used as a cost effective alternative to erythromycin for paediatric cholera, especially in children with concomitant acute respiratory infection. PMID- 9850411 TI - Serological similarities between hyperreactive malarial splenomegaly and splenic lymphoma in west Africa. AB - In West Africa hyperreactive malarial splenomegaly (HMS) and splenic lymphoma with villous lymphocytes (SLVL) are demographically and clinically indistinguishable. Determination of lymphocyte clonality is needed to differentiate clearly between these 2 disorders. To obtain evidence to support our hypothesis that HMS and SLVL are aetiologically related we studied the serological profile of malaria-related antibodies in HMS and SLVL in West Africa. We found that in SLVL total immunoglobulin M and antimalarial antibody levels were markedly raised, a combination which is characteristic of HMS. These findings strongly support a developmental relationship between HMS and SLVL in tropical Africa and implicate malaria in this process. PMID- 9850412 TI - Inhibition of growth in vitro of Plasmodium falciparum by vitamin E (alpha tocopherol). PMID- 9850413 TI - Pathophysiology of Ebola haemorrhagic fever. PMID- 9850414 TI - Dried blood spot technique for detecting Treponema infection. PMID- 9850416 TI - The strategy of Myxococcus xanthus for group cooperative behavior. AB - New evidence has been presented from our laboratory that the gliding bacterium, Myxococcus xanthus, does not home by chemotaxis toward a nutrient source. Our experiments, those of others, and the theory presented here combine to suggest a model, called the 'Pied Piper' model. It hypothesizes a gene that has a high mutation rate forward and back (say something in excess 10(-4) mutations per cell generation) which leads to switching between two motility states. Occasionally rare organisms become genetically, but reversibly, changed so that they move unidirectionally instead of mostly forward and back as do the bulk of the cells. When such a 'leader' cell arises, it continues to move in its original orientation, and causes a cohort of cells to move together away from the bulk of the cells. That is, in the less common mutational state it counteracts the usual tendency to just move forward and backward achieving little net movement. The assumption of a genetic element that mutates in a reversible way is suggested by numerous cases of reversible switches now known in a wide range of bacteria serving a variety of functions. A second aspect of the model is that mechanisms exist that cause cells to move in the same direction as their nearby neighbors. This process results in a regular spacing of bands of cells to form mounds in the absence of a leader. The action of C-factor, a factor-secreted by the cells which has been largely studied in the laboratory of Dale Kaiser, and extracellular fibrils, (rod-shaped protein and carbohydrate bodies) largely studied in the laboratory of Martin Dworkin, may be key elements in coordinating (or linking) the movements of neighboring cells. Based on the assumption of the absence of chemotaxis, computer simulations of pattern formation for gliding bacterial swarms and flares are consistent with observed behaviors and thus are additional evidence that chemotactic motility of the type exhibited by Escherichia coli, is not necessary for the group movements of M. xanthus. Some tests for this model are suggested. PMID- 9850415 TI - Biosynthesis of glycoproteins in Candida albicans: solubilization and partial characterization of dolichol phosphate mannose synthase and protein mannosyl transferases. AB - Incubation of a mixed membrane fraction isolated from C. albicans yeast cells with Nonidet P-40 at a detergent/protein ratio as low of 0.025 (0.016-0.019%, w/v) yielded a soluble fraction that catalyzed the transfer of mannose from GDP [14C] Man into dolichol phosphate mannose and from this intermediate into mannoproteins. Over 95% of the sugar in mannoproteins was O-linked as judged from its release after beta-elimination. Mannose was identified as the sole product after this treatment. Transfer activity did not depend on exogenous lipid acceptor indicating that the latter was solubilized along with the mannosyl transferases. Synthesis of mannolipid and mannoproteins occurred at optima temperatures of 20 degrees C, and 37 degrees C, respectively, and at a pH in the range of 7.5-9.5. Mannosyl transfer into the mannolipid was stimulated by Mg2+ and inhibited by Ca2+ and Mn2+ whereas mannoprotein labeling was stimulated by Mn2+ and to a lower extent by Mg2+. When measured as a function of substrate concentration, the synthesis of the mannolipid was a nearly linear function of GDP-Man concentration in the range of 5 to 32 microM whereas protein mannosylation exhibited hyperbolic kinetics with saturation reached at about 10 microM. The solubilized preparation was able to utilize an exogenous source of mannolipid as sugar donor for protein mannosylation. Dinucleotides and, to a higher extent trinucleotides, inhibited mannosyl transfer into the mannolipid and hence into mannoproteins. PMID- 9850417 TI - Decreased susceptibility of Malassezia furfur to UV light by synthesis of tryptophane derivatives. AB - Recently, tryptophane (Trp)-dependent synthesis of pigments and fluorochromes in Malassezia furfur was described. The possible significance of this metabolic pathway for the microorganism remains to be explored. Since the upper parts of the human epidermis are a natural habitat of M. furfur, increased exposure to UV light may be hazardous. Five reference strains and one wild type strain of M. furfur were grown on m-Dixon agar, in which the nitrogen source peptone had been substituted either by pigment-inducing tryptophane or arginine. The yeast cells thus obtained were harvested after 6 days, washed with physiological saline and inoculated on to the modified Dixon medium. Immediately after inoculation, the yeast cells were irradiated with UVA (100, 150 and 200 Jcm-2, single dose) or UVB (100, 500, 1000, 1500, 2000 mJcm-2, single dose; 500, 1500, 2500 mJcm-2 cumulative dose). Irrespective of the primary nitrogen source (Trp or Arg), unexposed controls showed nearly identical cell yield after 5 days. In the case of irradiation, however, growth reduction of cells cultured on Trp was lesser than that of cells fed with arginine. High significance (p < 0.0001) was found especially with the upper UVA and UVB doses. Differences were also found among the individual test strains, the wild strain being most sensitive. One strain (CBS 6094) failed to produce pigment on Trp medium, and there were no differences in the growth behavior of subcultures of this strain fed with either arginine or tryptophane under irradiation. In conclusion, synthesis of pigments and fluorochromes by M. furfur implies the generation of potent UV filters in the UVA and UVB spectrum. PMID- 9850418 TI - Metabolic characterization of Kloeckera apiculata strains from star fruit fermentation. AB - A total of 37 strains of Kloeckera apiculata was isolated during the spontaneous fermentation of star fruit must. Each strain was differentiated from the others on the basis of its capacity to produce acetaldehyde, ethyl acetate, higher alcohols, acetoin and acetic acid. All the strains were characterized by the low production of higher alcohols and the high production of ethyl acetate, whereas consistent differences in the production of acetaldehyde, acetoin and acetic acid served to differentiate star fruit apiculate strains into six different phenotypes, present at different stages of the fermentation process. The metabolic strain diversity found can be interpreted as a natural consequence of environmental conditions, which influenced the frequency and selection of specific apiculate strains. From the biotechnological point of view the different metabolic biotypes represent an important source of strains for potential use as starter cultures for star fruit fermentation. PMID- 9850419 TI - Inter-strain comparison by pyrolysis mass spectrometry of Staphylococcus aureus isolates associated with nosocomial infection. AB - Pyrolysis mass spectrometry was used to characterise Staphylococcus aureus isolates from an outbreak of postoperative wound infections on a mixed surgical ward. The PyMS results were compared with those of phage typing. Both suggested a single strain of S. aureus, of phage type 3C, 55, 71, was responsible for six of the 13 wound infections. PyMS differentiated an isolate from a member of staff of similar phage type to the epidemic strain, which had previously been considered to be the point source for the outbreak. PyMS is a rapid and inexpensive technique for investigating nosocomial outbreaks, including those caused by S. aureus and, in this instance, was more discriminatory than phage typing. PMID- 9850420 TI - Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. AB - Approximately 500 species of ascomycetous yeasts, including members of Candida and other anamorphic genera, were analyzed for extent of divergence in the variable D1/D2 domain of large subunit (26S) ribosomal DNA. Divergence in this domain is generally sufficient to resolve individual species, resulting in the prediction that 55 currently recognized taxa are synonyms of earlier described species. Phylogenetic relationships among the ascomycetous yeasts were analyzed from D1/D2 sequence divergence. For comparison, the phylogeny of selected members of the Saccharomyces clade was determined from 18S rDNA sequences. Species relationships were highly concordant between the D1/D2 and 18S trees when branches were statistically well supported. PMID- 9850421 TI - Biosynthesis of glycoproteins in Candida albicans: biochemical characterization of dolichol phosphate glucose synthase. AB - A mixed membrane fraction isolated from C. albicans yeast cells catalyzed the transfer of glucose from UDP-Glc into three classes of endogenous acceptors: glucolipid, glycoprotein and lipid-linked oligosaccharides. About 80% of the total radioactivity transferred into these products corresponded to the glucolipid which was identified as dolichol phosphate glucose by several criteria. The remainder was detected in about equal proportions in the other two fractions. Conditions that stimulated or inhibited glucolipid synthesis did not affect the extent of glycoprotein labeling. The synthesis of dolichol phosphate glucose exhibited a K(m) of 104 microM UDP-Glc and was stimulated by Mg2+ but not by Mn2+ or Ca2+. The latter cations were, however, better stimulators of glycoprotein labeling than Mg2+. Most nucleotides strongly inhibited the synthesis of dolichol phosphate glucose, UMP being a competitive inhibitor with a Ki of 100 microM. The dolichol phosphate glucose synthase reaction was reversed about 57% by 0.62 mM UDP but not by UMP. PMID- 9850422 TI - [Drug research in Germany--problems and chances--from the viewpoint of the drug industry]. AB - A trend can be observed for multinational research-based drug companies shifting their resources away from Germany (and partly also from Europe) and expanding their preclinical and clinical drug research and development programmes and organizations in the USA. The main reasons, which are presently under discussion in Germany, are the image and public acceptance of clinical drug research, professionalism of clinical studies (i.e. availability of an adequate infrastructure), the process of ethics committees votes, the citation impact of publications, the legal obstacles, the implementation of Good Clinical Practice (GCP), the regulatory and institutional situation and the political environment. These problems have been recognised and appraised recently by the pharmaceutical industry, universities, scientific associations, and governmental institutions. Prerequisites to improve the basic conditions of clinical research in Germany to make it internationally competitive are proposed in this paper. PMID- 9850423 TI - Inhibition of R-[3H]-baclofen binding to rat brain synaptic membranes by a homologous series of phenyl alcohol amides anticonvulsants and their evaluation as GABAB receptor blockers. AB - The inhibition of the specific R-[3H]-baclofen binding to GABAB (gamma aminobutyric acid) sites by a homologous series of phenyl alcohol amides was tested in rat brain synaptic membranes. Some of these phenyl alcohol amides were designed as anticonvulsants as well as antagonists of the GABAB receptor. These anticonvulsants showed a high affinity to the GABAB receptor. DL-2-hydroxy-2-(4' chlorophenyl)butyramide and DL-3-hydroxy-3-(4'-chlorophenyl)pentanamide (DL-Cl HEPP) were as effective as GABA and R-baclofen and were the most potent examined. DL-2-hydroxy-2-phenylbutyramide (DL-HEPA), (+)-HEPA, (-)-HEPA, DL-3-hydroxy-3 phenylpentanamide (DL-HEPP) and DL-4-hydroxy-4-phenylhexanamide (DL-HEPB) were as potent as DL-baclofen. The phenyl alcohol amides with fluorine in the para position of the phenyl ring were less active than DL-2-hydroxysaclofen. DL-4 hydroxy-4-(4'-chlorophenyl)hexanamide was the least active of the series. In addition, R-baclofen antagonized very effectively the anticonvulsant activity of both DL-HEPP and DL-Cl-HEPP in a dose-dependent fashion. These results support the assumption that some of these phenyl alcohol amides anticonvulsants are GABAB receptor blockers. PMID- 9850424 TI - Cerebral blood flow effects of sodium valproate in drug combinations in the baboon model. AB - Sodium valproate (CAS 1069-66-5, Epilim) has been used in the management of epilepsy during the last three decades. Although important information on the pharmacological actions and efficacy of sodium valproate has accrued to date, limited research has been conducted on its effects on cerebral blood flow. In recent years, with the aid of SPECT (single photon emission computed tomography) and PET (positron emission tomography) it has been shown that marked cerebral blood flow changes occur in epileptic patients. Furthermore it was established recently that sodium valproate influences the cerebral blood flow in children by decreasing the flow significantly. The present study investigated the effects of sodium valproate on the cerebral blood flow, using 99mTc-HMPAO (hexamethylpropylene amino oxime) and SPECT, in a primate model, as well as the effects of its drug interactions with therapeutic agents that influence cerebrovascular dynamics, e.g. sumatriptan, nimodipine and acetazolamide. The current study using single dose treatment with sodium valproate did not detect a decrease or increase of the cerebral blood flow when compared with control baseline results. Drug interaction between sodium valproate and nimodipine may occur as a reduction of 25% in cerebral blood flow from the baseline control was observed in this case. The effects observed for the combinations of sodium valproate respectively with sumatriptan and acetazolamide are attributed to the influences of the sumatriptan (decrease) and acetazolamide (increase) alone. The cerebral blood flow effects of these drugs and possible interactions during an acute epileptic seizure need to be investigated. PMID- 9850425 TI - Pharmacokinetics of deramciclane in rabbits. AB - The pharmacokinetics of deramciclane (CAS 120444-78-8, EGIS-3886) was investigated in rabbits after i.v., p.o. and s.c. administration of 3 mg/kg 14C phenyl-deramciclane. The plasma, concentration-time curves of total radioactivity, the parent compound (deramciclane) and its N-demethylated metabolite (EGIS-7056) were determined. The radioactivity level was measured by liquid scintillation technique while the concentration of the parent compound and its metabolite was determined by gas chromatography-mass spectrometry detection. The p.o. and i.v. studies were carried out on the same group of animals, while a separate group of rabbits was used for studying s.c. absorption. Deramciclane was readily absorbed after p.o. and s.c. treatment (tmax 1.0 to 1.4 h). The terminal elimination half-life (t1/2 beta) of the parent compound fell between 5.8 to 7.1 h, while that of the total radioactivity ranged from 21.6 and 26.0 h. The absolute bioavailability of deramciclane calculated from the AUC0-infinity values was found to be 43 and 60% after p.o. and s.c. treatment. The apparent volume of distribution (Vd) and the whole body clearance (Cl) of deramciclane after i.v. administration were 25.0 +/- 7.1 l/kg and 2.6 +/- 0.5 l/h/kg, respectively. The AUC0-infinity values of the parent compound varied between 4.6 and 7.9% of that of total radioactivity, suggesting that deramciclane was subjected to intensive metabolic conversion. The AUC0-infinity of N-desmethyl-deramciclane was 5.7%, compared to that of the parent compound after i.v. administration. PMID- 9850426 TI - Pharmacokinetics of phenprobamate after oral administration to healthy subjects. AB - Phenprobamate (CAS 673-31-4) is a centrally acting skeletal-muscle relaxant agent. There are only two studies in the literature about the pharmacokinetics of phenoprobamate in man. The inconsistency between the results of these studies can be attributed partly to the different analytical methodologies used. A sensitive, specific and reproducible HPLC-assay, which may increase the reliability of the pharmacokinetic studies of phenprobamate in plasma, has been developed recently. The objective of this investigation was to assess the single-dose kinetics of phenprobamate in human and to determine the pharmacokinetic parameters of clinical and regulatory concern. The plasma pharmacokinetics of phenprobamate have been investigated following single oral administration at a dose of 800 mg in eleven healthy volunteers. PMID- 9850427 TI - [The bioequivalence of a new amitriptyline tablet formulation in comparison with a reference preparation]. AB - An investigation of the bioequivalence of a new tablet formulation (amitriptylin 25 von ct) with 28.3 mg amitriptyline hydrochloride (CAS 549-18-8) was performed in a two-way cross-over study with 18 subjects. The relative bioavailability with respect to a reference preparation for AUC related to amitriptyline (CAS 50-48-6) was 99.3% and for Cmax 100.4%. A positive decision for bioequivalence derived from the usual confidence intervals for both parameters related to amitriptyline and the metabolite nortriptyline (CAS 72-69-5), respectively tmax showed no difference. The new formulation was bioequivalent to the reference. PMID- 9850428 TI - Studies of the cardioprotective effects of ascorbic acid in isolated rabbit hearts. AB - Ascorbic acid (CAS 50-81-7) might mediate cardioprotective effects by scavenging free oxygen radicals. The effects of exogenous ascorbic acid on acute myocardial ischemia (MI) was investigated in isolated electrically-driven rabbit hearts (Langendorff, constant pressure: 70 cm H2O, Tyrode solution, Ca2+ 1.8 mmol/l, 37 degrees C). Repetitive MI, separated by a reperfusion period of 50 min, was induced by coronary artery branch ligature and quantitated from epicardial NADH fluorescence photography. Starting after a reperfusion period of 20 min, isolated hearts were treated with ascorbic acid (10(-5) or 10(-4) mol/l). Ascorbic acid had no significant influence on the left ventricular left ventricular pressure or the coronary flow (p > 0.05). Ascorbic acid had no significant effect on epicardial NADH-fluorescence area or intensity (p > 0.05). Free radical scavenging properties reported for ascorbic acid do not mediate cardioprotective effects at the concentrations used in isolated rabbit hearts. PMID- 9850429 TI - Protective effects of delapril, indapamide and their combination on stroke occurrence and lifespan in salt-loaded stroke-prone spontaneously hypertensive rats. AB - The effects of long-term oral administration of delapril (CAS 83435-67-0), indapamide (CAS 26807-65-8) and their combination on the occurrence of stroke and on mortality were investigated in young salt-loaded stroke-prone spontaneously hypertensive rats (SHRsp) for 31 weeks of treatment (8th-39th week of age) and up to 8 weeks thereafter. Body weight and saline consumption were investigated at regular intervals and cerebrovascular lesions, renal and heart weight were assessed after sacrifice. Untreated SHRsp served as controls. About 50% of control animals died within 6 weeks of saline administration and in 56% of surviving animals cerebral lesions were present at sacrifice, while no death and no cerebral lesions were observed in animals drinking saline, to which delapril, indapamide and their combination had been added, up to the end of treatment. This protective effect was maintained even in the withdrawal period. All treatments induced a highly significant (p < 0.001) reduction of heart weight/body weight and kidney weight/body weight ratios. PMID- 9850430 TI - Pharmacokinetics of a single dose of phenylpropanolamine following oral administration at two different times of the day. AB - Pharmacokinetics of a single oral dose of phenylpropanolamine (CAS 154-41-6) was investigated by administering 50 mg of the drug at 10.00 and 22.00 h to 8 healthy male volunteers in a crossover design with a wash-out period of 10 days. Serum samples were analysed for phenylpropanolamine using high performance liquid chromatography. Pharmacokinetic parameters were calculated using model independent method. A significant (p < 0.05) elevation in Cmax (227.45 versus 181.98 micrograms/l) was observed following the drug administration at 22.00 h as compared to 10.00 h. These variations may be due to circadian changes in gastric pH contributing to the time dependent changes in the absorption of the drug. PMID- 9850432 TI - Pharmacokinetic characteristics of the novel anticancer agent CPT-11 and its active metabolite in plasma and lung lymph fluid following intravenous administration to sheep. AB - 7-Ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin (CPT-11, 100286-90-6) is one of the most promising novel anticancer agents, especially for lung cancer. 7-Ethyl-10-hydroxycamptothecin (SN-38), an active metabolic of CPT 11, has much stronger cytotoxicity than CPT-11. The present study was designed to evaluate the distribution and behavior of CPT-11 and SN-38 in lung lymph circulation following intravenous infusion. Awake sheep with chronically instrumented lung lymph fistulas were prepared. The concentrations of CPT-11 and SN-38 in plasma and lung lymph fluid were measured after intravenous infusion of 100 mg/body of CPT-11 for 90 min. SN-38 constantly showed higher lymph to plasma concentration ratios than those of CPT-11, and the % area under the curve (AUC) ratio of SN-38/CPT-11 in lymph fluid was significantly higher than that in plasma. These data indicated that SN-38 distributed in lung tissue moved more easily into lung lymph fluid than CPT-11, and might be more rapidly metabolized in lung tissue than plasma. CPT-11 may have favorable therapeutic effects on intrathoracic malignancies such as lung cancer and lymph metastasis. PMID- 9850431 TI - Determination of the active moiety of BX661A, a new therapeutic agent for ulcerative colitis, by studying its therapeutic effects on ulcerative colitis induced by dextran sulfate sodium in rats. AB - 5-[4-(2-Carboxyethylcarbamoyl)phenylazo]salicylic acid disodium salt dihydrate (CAS 80573-04-2, BX661A) is being developed as a therapeutic drug for ulcerative colitis. To determine the active therapeutic moiety of BX661A, the therapeutic effects with single and combined administration of 5-aminosalicylic acid (5-ASA), 4-aminobenzoyl-beta-alanine (4-ABA) and 4-amino-N-2-pyridinyl-benzenesulfonamide (CAS 144-83-2, sulfapyridine, SP) on ulcerative colitis induced by dextran sulfate sodium (DSS) in rats were investigated, and the following results were obtained. 1. BX661A at doses of 30, 100 and 300 mg/kg (p.o.) dose-dependently decreased the erosion area (mm2) in the large intestine with % inhibition values of 28.7, 49.1 and 61.6%, and the shortening of the large intestine with % inhibition values of 17.1, 25.7 and 48.6%, respectively. Salazosulfapyridine (SASP) at doses of 30 and 100 mg/kg (p.o.) decreased the erosion area (mm2) in the large intestine with % inhibition values of 30.7 and 45.3%, respectively, but did not improve the shortening of the large intestine. However, at a dose of 300 mg/kg (p.o.) SASP, the % inhibition value of the erosion area in the large intestine was reduced. 2. A single intrarectal administration of 5-ASA (105 mg/kg, i.r.) significantly decreased the erosion area (mm2) in the large intestine, but a single administration of 4-ABA or SP did not show any significant effect on the erosion area. Combined administration with 5-ASA (105 mg/kg, i.r.) and 4-ABA (142.8 mg/kg, i.r.) significantly decreased the erosion area (mm2) in the large intestine with a % inhibition value of 63.8%. On the other hand, the efficacy of 5-ASA disappeared with combined administration with SP (% inhibition value of 7.3%). These results suggest that 5-ASA is the active moiety for the therapeutic effects of BX661A and indicate that the efficacy of 5 ASA disappears with the combined use of SP, but not of 4-ABA. Therefore, it seems that BX661A is clinically safe and more effective than SASP in the treatment of patients with ulcerative colitis. PMID- 9850433 TI - Effects of imipenem and meropenem on serum sensitivity and surface hydrophobicity of Klebsiella pneumoniae. AB - Serum sensitivity and surface hydrophobicity of two Klebsiella pneumoniae strains (internal strain No. 378 and 3259) exposed to imipenem (CAS 64221-86-9) and meropenem (CAS 96036-03-2) at subinhibitory concentrations (sub-MICs; 1/4, 1/8 and 1/16 of the MICs) were evaluated. Carbapenems at all sub-MICs tested (with the exception of 1/16 of the MICs in strain 378) decreased susceptibility of bacteria to serum bactericidal activity. All sub-MICs of the antibiotics tested also reduced the bacterial surface hydrophobicity. The surface hydrophobicity of strain 3259 was most effectively decreased after the exposure to imipenem and meropenem at 1/4 of the MICs (to 3% or 5.2% of the control values). The highest decrease of hydrophobicity in strain 378 was found after exposure to imipenem and meropenem at 1/16 of the MICs (19.2% or 32.3%). PMID- 9850434 TI - Softgel capsule technology as an enhancer device for the absorption of natural principles in humans. A bioavailability cross-over randomised study on silybin. AB - In order to evaluate if a patented soft gelatine capsule could improve the bioavailability of silybin (CAS 22888-70-6) in comparison to a hard shell capsule, an open, single dose, two-way, balanced cross-over study, was performed. The study was conducted on 12 healthy subjects (6 M and 6 F). 80 mg of silybin in a 1:2 complex with phosphatidylcholine was administered. Blood was sampled from the subjects in two occasions at the following times after drug administrations: 0 (sample before dosing), 1, 2, 3, 4, 6 and 8 h. The pharmacokinetic parameters calculated from the results of the plasma analyses demonstrated that the mean values of both Cmax and AUC0-1 were increased when the patented soft gelatine capsule formulations were administered (i.e. Cmax more than 3-fold and AUC0-1 more than 2-fold). PMID- 9850435 TI - Somatic gene therapy. Present situation and future perspective. AB - The ultimate goal in the management of inherited as well as acquired diseases is a rational therapy with the aim to eliminate the underlying biochemical defects, rather than a symptomatic treatment. Among other approaches somatic gene therapy is a promising candidate to meet these objectives and appears to have the potential to revolutionize modern medicine. Gene therapy is characterized by the transfer of genetic information to a patient through the use of recombinant DNA technology. Several strategies for the treatment of monogenetic disorders as well as chronical diseases like cancer and AIDS have been used in various somatic gene therapy projects. So far, 329 clinical studies (phases I, I/II and II) with over 2500 patients have been initiated worldwide since 1989. No significant toxicity and adverse side effects have been observed. To allow efficient transfer of the therapeutic genes, a variety of gene delivery techniques have been developed based on viral and non-viral vector systems. For the success of this technology it is vital to achieve regulated and sustained expression of foreign genes in specific target tissues. This will be crucial for the widespread application of somatic gene therapy. So far none for the gene delivery systems is able to meet the requirements of safety, efficiency and specificity demonstrating that vector research will be an important focus in the development of optimized transfer methods. From a regulatory point of view pharmaceutical DNA-products can be regarded as drugs and are therefore subject to the same regulations. Human gene therapy must, however, be limited to manipulations affecting somatic, differentiated cells to prevent the transferred gene from being transmitted to the individual's descendants. Applications for the purpose of 'enhancement' and not for the treatment of diseases are also not acceptable. Under these prerequisites, somatic gene therapy does not raise any new ethical concerns and can be interpreted as a special form of an organ transplantation. A comparison of the different regulatory situations of gene therapy in Europe and the United States demonstrates that for the European countries a uniform regulation is desired. Today somatic gene therapy is still in its infancy. It will continue to be scientifically and technically challenging until simple and effective procedures will have been developed. Demonstration of its clinical efficacy especially in the long term will have to be the next step. Looking at the history of biotechnology and the success of the biotechnology industry that is now providing safe and efficient products from recombinant DNA-technology there is little doubt that gene therapy will become a successful treatment for various indications in the next decade. The purpose of this article is to review the current status of the development in somatic gene therapy. PMID- 9850436 TI - The pharmacological properties of lipophilic calcium antagonists. AB - Several types of calcium antagonists (CA) (verapamil, diltiazem, nifedipine and related drugs) may be used as antihypertensives. In practice, the dihydropyridines (nifedipine and related drugs) are the CA used most frequently as antihypertensives. Apart from the lowering of blood pressure CA may lead to other, theoretically beneficial, effects: regression of left ventricular and vascular hypertrophy, renal protection, weak natriuretic, weak antiplatelet, anti ischaemic and antiatherogenic activity. Several new dihydropyridine CA have been introduced in recent years. The advantages of the newer compounds, such as amlodipine, felodipine, isradipine, lacidipine and lercanidipine, may include: vasoselectivity, hence little or no cardiodepressant activity; an improved kinetic profile, resulting in a slow onset and long duration of action, fewer side-effects such as reflex tachycardia and headache, owing to the slow onset of the antihypertensive action. For a few newer CA a predominant effect on specialized circulatory beds (renal, coronary and cerebral) has been claimed. The new CA, which are clearly lipophilic, deserve special attention. Owing to the lipophilic character of such compounds considerable concentration occurs in lipid containing membrane depots. The CA thus concentrated are slowly released from these depots and, subsequently, reach their targets, the L-type calcium channels. This phenomenon explains both the slow onset and the long duration of action of these CA. Owing to the slow onset of action reflex tachycardia is virtually absent. The long duration of action allows satisfactory control of blood pressure in hypertensives by means of a single daily dose. A few lipophilic dihydropyridine CA are vasoselective. This property implies that at therapeutic, vasodilatory dosages no cardiodepressant activity occurs. Lercanidipine is a recently introduced example of a lipophilic and vasoselective dihydropyridine CA. It is an effective vasodilator/antihypertensive drug, with a slow onset and long duration of action, which is associated with neither reflex tachycardia nor cardiodepressant activity. Other examples of recently introduced lipophilic CA are lacidipine, barnidipine and manidipine. PMID- 9850438 TI - The new calcium antagonist lercanidipine and its enantiomers affect major processes of atherogenesis in vitro: is calcium entry involved? AB - Atherosclerosis results from multiple factors and involves several mechanisms, including endothelial monocyte and smooth muscle cell (SMC) changes, cholesterol accumulation, plaque rupture and thromboembolism. Calcium ions play a role in the initial and chronic development of atherosclerotic lesions. Several studies in experimental animal models have demonstrated the potential direct antiatherosclerotic effects of calcium antagonists. In this study the antiatherogenic activity of lercanidipine, a new lipophilic, second-generation calcium antagonist, was investigated. Lercanidipine and its enantiomers inhibited the replication and migration of arterial myocytes in concentrations ranging from 10 to 50 microM. The antiproliferative effect of lercanidipine was dose dependent, with a potency similar to that of lacidipine and nifedipine, and was unrelated to the stereoselectivity of enantiomers to bind L-type calcium channels. Lercanidipine and its enantiomers (25 microM) decreased the serum induced elevation of [Ca2+]i in SMC, with the (S)-enantiomer (69% inhibition) being 2.4-fold more active than the (R)-counterpart (29% inhibition). The studies performed with enantiomers of lercanidipine suggest that the observed effects are not related to the blockade of voltage-dependent Ca2+ channels and confirm, at least in vitro, the pharmacological potential of the compound to influence negatively the process of atherogenesis. PMID- 9850437 TI - Lercanidipine: short plasma half-life, long duration of action and high cholesterol tolerance. Updated molecular model to rationalize its pharmacokinetic properties. AB - Calcium-channel antagonist drugs of the 1,4-dihydropyridine type have been shown to bind to the L-type calcium channel. These drugs are not only amphiphilic, but new molecular designs have become increasingly lipophilic and can readily transport across cell membranes, accessing both hydrophilic and hydrophobic environments, despite becoming more soluble in the membrane bilayer. This biophysical understanding appears not only to define the molecular pathways for drug binding to the calcium-channel receptor, but also to explain differences in the overall clinical pharmacokinetics observed for different drugs in this class. The pharmacokinetic profile of calcium antagonists, although influenced to some degree by interactions with their target calcium-channel receptor, appears to be largely dictated by their interactions with cell membranes at the molecular level. There appears to be a correlation between the duration of action of such membrane-active drugs and the membrane partition coefficient in conjunction with the washout rate. This class of drugs has evolved from a drug such as amlodipine, with a long duration of action related to prolonged plasma half-life, to lercanidipine, which has the shortest plasma half-life relative to its intrinsically long duration of action. Recently, it was discovered that membrane cholesterol reduces the amount of calcium-channel antagonist that can partition into the membrane. Atherosclerotic disease results in increased levels of membrane cholesterol in smooth muscle cells. Latest generation calcium antagonist, which have a long duration of action, can better overcome this negative effect. Lercanidipine has now been shown to have one of the highest measured tolerances to cholesterol, which may indicate its ability to treat a broad range of hypertensive patients with varying degrees of progressive atherosclerotic disease. On what criteria should the effectiveness of calcium antagonists be evaluated? A good calcium antagonist needs to exhibit a placebo like side-effect profile, thus ensuring good patient compliance. However, an intrinsically long-lasting, once-a-day dose is also pharmacokinetically desirable. To be a truly optimal calcium antagonist, it should function and be efficacious over a broad range of hypertensive patients. It should be able to control blood pressure in light of other complications such as progressive atherosclerotic disease. Recent studies indicate that during the progression of atherosclerosis, cholesterol levels within cell membranes of the arterial wall increase, a process that can reduce the effective concentration of calcium antagonists in these membranes. What is needed is a calcium antagonist that is slow acting to reduce vasodilatory induced side-effects and intrinsically long lasting to ensure once-a-day dosage, and that possesses a high cholesterol tolerance factor to overcome the molecular and compositional changes taking place in the arterial wall, so that it can treat effectively a broad range of hypertensive patients. PMID- 9850439 TI - Clinical advantages of lipophilic dihydropyridines. AB - Lipophilic dihydropyridines have many theoretical and practical clinical advantages owing to their long permanence at the cell membrane. They have a greater chance of smoothly and permanently reducing blood pressure over 24 h than other dihydropyridines, a feature that may have positive prognostic implications since 24-h blood pressure is more closely related to the end-organ damage of hypertension. They may avoid the sympathetic activation consequent to an excessive early-dose hypotension, which is responsible for an increase in 24-h blood-pressure variability and reflex tachycardia, two phenomena that may worsen the prognosis of hypertensive patients. A further advantage which has been shown in experimental and clinical settings is the possibility of reducing the extension and progression of atherogenic lesions in blood vessels, which are responsible for cardiovascular complications in hypertension. Some of these features have been shown by the novel lipophilic dihydropyridine lercanidipine. In particular, clinical studies have shown that (i) this drug is effective in homogeneously reducing blood pressure over 24 h, (ii) its antihypertensive effect is similar to that of some common antihypertensive drugs, and (iii) the rate of adverse events experienced with lercanidipine is no greater than that observed with other antihypertensive drugs, with special reference to non-lipophilic calcium antagonists. In particular, studies performed so far have shown that lercanidipine does not exert a dangerous reflex tachycardia. PMID- 9850440 TI - The role of tacrolimus in adult kidney transplantation: a review. AB - The use of tacrolimus (FK506) in adult kidney-transplant recipients has been the subject of a number of single- and multi-center studies. This review article focuses on those studies in which tacrolimus was used either as rescue therapy in patients who developed refractory rejection on cyclosporine (CyA)-based regimens or as primary immunosuppression in adult renal-allograft recipients. Twenty-five prospective and retrospective studies conducted in the US, Japan and Europe, including single- and multi-center experiences, were identified in the medical literature. Of these studies, most show a 74-98% initial success rate for tacrolimus rescue therapy. Comparative studies reviewed herein demonstrate comparable patient- and graft-survival rates between tacrolimus- and CyA-treated patients. Many studies have shown that rejection episodes occur with similar or lower frequency among patients treated with tacrolimus than among those given CyA as primary immunosuppression. The major toxicities associated with tacrolimus are nephrotoxicity, neurotoxicity and diabetogenicity. Results from several studies have also demonstrated an association between these tacrolimus side effects and high whole-blood trough levels of tacrolimus. In many cases, a reduction in dosage can reverse these adverse effects. In summary, based on both single- and multi-center data, tacrolimus has been demonstrated to be efficacious when used for either primary immunosuppression or as rescue therapy for refractory acute rejection in adult renal-allograft recipients. PMID- 9850441 TI - Donor cerebral tissue pulmonary emboli in a functioning transplanted lung. AB - Cerebral tissue pulmonary embolus (CTPE) is a rare event following severe blunt or penetrating head trauma and is often complicated by coagulation disturbances and hemorrhage. Donor cerebral tissue pulmonary embolism has been reported to cause lethal, early graft dysfunction in lung transplant recipients. We report a case of donor cerebral tissue pulmonary embolism in a 41-year-old female single lung transplant recipient with excellent post-operative graft function. PMID- 9850442 TI - Hepatic artery pseudoaneurysm with hemobilia following angioplasty after liver transplantation. AB - A 58-yr-old female with primary biliary cirrhosis underwent an uncomplicated orthotopic liver transplantation. Elevated liver function tests 2 months post transplantation were evaluated with Doppler ultrasound and a hepatic artery stricture was documented. The hepatic artery stenosis was treated with angioplasty. She developed hemobilia 1 d after the procedure, which was confirmed by angiography. Emergent exploratory laparotomy revealed a pseudoaneurysm at the hepatic artery anastomosis. The pseudoaneurysm was resected and the proper hepatic artery of the graft was anastomosed to the splenic artery of the host using preserved homograft. Her post-operative course was uneventful and liver function tests returned to normal quickly after the surgery. This report will discuss the unusual nature of this complication, and review the problem of hemobilia and pseudoaneurysms in liver transplant recipients. PMID- 9850443 TI - Effect of recipients' peri-operative parameters on the outcome of kidney transplantation. AB - In this prospectively collected data base multifactorial study, the influence of the peri-operative recipients' parameters on the transplanted kidney function were examined. One hundred and twenty-one patients who underwent first cadaver kidney transplantation in our unit were involved and were followed up for another 2 yr. The recipients were divided into three groups. Group I, patients with non functioning grafts, group II, with delayed graft function, and group III, with good graft function. All routinely measured parameters were involved in the examination. The pre-operative haematocrit level differed significantly among the three groups. Significant intra-operative differences were found among the three groups in mean arterial pressure, (group I: 81 +/- 31 mmHg, group II: 90 +/- 24 mmHg, vs. group III: 108 +/- 26 mmHg, p < 0.01) at 5 min before revascularisation. The patients who had better post-operative kidney function, had lower pre-operative haematocrit level, and higher blood pressure in the different phase of operation, than those who had post-operative kidney function problems. The second warm ischaemic time in Group III was significantly shorter than in group I. (Group I: 51.5 min vs. group II: 47 min, vs. group III: 46.3 min, p < 0.001). Rejection rate was higher during the first 5 post-operative days in patients with non-functioning grafts. (Group I: 53% and group II: 24% vs. group III: 12% p < 0.001). The other examined parameters had not differed significantly among the three groups. The kidney function differences observed in the early (first 5 d) post-operative period, remained similar at the end of the next 2 yr. According to our results the peri-operative fluid-balance is one of the most important factors which may influence the success of the kidney transplantation. PMID- 9850444 TI - Development of Sang-35: a cyclosporine formulation bioequivalent to Neoral. AB - During a research program, SangStat Medical Corporation developed more than 270 oral cyclosporine formulations. On the basis of animal and clinical trials, Sang 35 was chosen for clinical development, and bioequivalence with the cyclosporine microemulsion Neoral was established. In a cross-over study involving 36 healthy male volunteers, single 500 mg cyclosporine doses of Sang-35 (AUC0-infinity: 13,900 +/- 2470 micrograms.h.L-1, mean +/- standard deviation (SD)) and of Neoral (AUC0-infinity: 14,000 +/- 2900 micrograms.h.L-1) resulted in equal areas-under the-time-concentration curve (AUC0-infinity). Sang-35 and Neoral were also bioequivalent in healthy male subjects after high-fat meals as well as in female and African-American subjects. In stable kidney transplant patients (n = 12) receiving a mean (+/- SD) cyclosporine dose of mg/d (3.6 +/- 1.6 mg/kg/d), AUC0 12 h after Sang-35 was, as expected, significantly higher than that after Sandimmune (4550 +/- 1858 vs 3468 +/- 1402 micrograms.h.L-1, p < 0.01). Sang-35 and Neoral resulted in equivalent cyclosporine AUC0-12 h values (4120 +/- 1508 and 4377 +/- 1579 micrograms.h.L-1, respectively) in stable kidney transplant patients (dose: 293 +/- 114 mg/d or 3.7 +/- 1.5 mg/kg/d, n = 32). In an additional study, 42 stable kidney graft patients were switched from Sandimmune to Sang-35. Based on a conversion strategy targeting AUC equivalence, only one dose adjustment was required in 55% of the patients, and 95% of patients (40 of 42) needed three or fewer dose adjustments. The mean Sang-35 dose was 7% lower than the mean Sandimmune dose. During the studies, Sang-35 and Neoral exhibited similar safety and tolerability profiles. It is concluded that Sang-35 and Neoral are bioequivalent and that patients can safely and easily be switched from Neoral or, in combination with dose adjustment, from Sandimmune to Sang-35. PMID- 9850445 TI - Trends in organ donation. AB - Renal and extrarenal transplant data were collected for seven geographical regions for the period 1989-1996. In Western Europe and North America the number of kidney donors increased by 926 and 2743, respectively. The total number of transplants also increased in both regions by 3756 and 6936, respectively. Renal transplants accounted for approximately 60% of the total number of transplants and, although the number of renal transplants did not alter in Western Europe, the number rose by 3055 in North America. Outside of these regions the number of extrarenal transplants was 3-18% of the total. The number of living kidney donors in North America increased each year and was higher than the number recruited in Western Europe (3389 vs 943 in 1996). With the exception of Eastern Europe, where virtually no renal transplants were carried out using organs from living donors, the number of living kidney donors rose in other regions: for example, in Latin America, the proportion of living kidney donors rose from 29% in 1970-88 to 51% in 1995, and, in Asia, 90% of kidneys were donated by living donors. As the quality of cadaveric donor organs is often sub-optimal, the use of living donors is likely to increase in both Western Europe and North America, but is unlikely to become the most important source of organs in these regions. PMID- 9850446 TI - Giving and taking--to whom and from whom? People's attitudes toward transplantation of organs and tissue from different sources. AB - The aims of the study were: 1) to describe the willingness of the public to receive material of different origins in one's own body; 2) to compare the willingness to donate and receive body material; 3) to compare the willingness to donate while alive and after death; 4) to compare the willingness to donate to a next-of-kin and unknown recipients. A random sample of 1500 inhabitants, 18 to 70 yr old, in the county of Uppsala, Sweden, were sent a questionnaire asking about their opinion on transplantation and transfusion issues. The response rate was 71%. Ninety-five percent accepted to receive blood transfusion, 89% bone-marrow transplantation, and 85% transplantation of a solid organ. Organs from living donors were preferred (77%), then organs from decreased donors (69%), then artificial organs (63%), and last animal organs (40%). More than half of those accepting transplants made exceptions for some types of organs. The youngest and those with higher education were more positive toward receiving all types of organs than the older ones and those with lower education. Women were less prepared than men to accept animal organs. Those who accepted organs from animals usually also accepted all other types of organs, and were willing to donate organs and tissue more often than those who did not accept to receive animal organs. The readiness to support a sick family member by giving bone-marrow and even a kidney was considerable, 89 and 81%, respectively. The attitudes were less positive with regard to giving blood and bone-marrow to unknown recipients, 54 and 41%, respectively. Sixty-one percent of the respondents were positive toward donating their own organs after death. Of those who were positive, 10% made exceptions for special organs that they did not want to donate, mostly heart, eyes, and brain. Individuals with higher education and young people were more often positive than those with lower education and old people regarding donation of blood and organs, and bone-marrow donation to a relative. Women were somewhat more accepting to donate while alive than males. Thirty-one percent, more often women than men, had signed a donor card and/or registered with the Swedish Organ Donation Registry. The results with regard to receiving organs and tissue are discussed in terms of two different sets of explanations, which can be seen as different sides of the same coin, and mutually strengthening the reactions. The great readiness to donate to a family member as well as the discrepancy between giving in life and after death is commented upon. PMID- 9850447 TI - The middle colic vein: an alternative source of portal inflow in orthotopic liver transplantation complicated by portal vein thrombosis. AB - BACKGROUND: Portal vein thrombosis (PVT) was previously considered a contraindication to orthotopic liver transplantation (OLT) since adequate portal blood supply is mandatory for graft function and patient survival. Improvements in surgical technique, however, have meant that this problem now can be circumvented in most instances. Nevertheless portal vein thrombosis remains an obstacle in OLT and is associated with increased incidence of primary non function and long-term liver failure. METHODS: A 55-yr-old patient underwent OLT for secondary biliary cirrhosis associated with hepatitis C infection and complicated by long standing PVT. Involvement of the portal, mesenteric, and splenic veins prevented standard portal venous reconstruction. Portal inflow was accomplished by a side-to-end anastomosis between the middle colic vein and the donor portal vein. RESULTS: Hepatic reperfusion and subsequent liver function were excellent. Portal blood flow, as measured by color-enhanced Doppler ultrasound, was normal following surgery until discharge. The post-operative course was complicated by abdominal wound dehiscence and recurrent cytomegalovirus (CMV) infection. The patient was discharged in good clinical condition, with excellent liver function and patent portal vein 89 d after OLT. CONCLUSIONS: The middle colic vein is a novel, not previously described, source of portal venous inflow for OLT complicated by extensive splanchnic venous inflow thrombosis. PMID- 9850448 TI - Acute rejection following renal transplantation. Evidence that severity is the best predictor of subsequent graft survival time. AB - Timely recognition of risk factors influencing graft survival time following kidney transplantation could facilitate the development of clinical interventions that would increase the longevity of graft survival. To identify these risk factors, we performed a retrospective analysis of the clinical outcome of 1949 renal transplants performed at The Ohio State University Transplant Program between 22 September 1982 and 14 June 1996. The number of acute rejection (AR) episodes was most predictive of shorter graft survival time. The first AR episode severity (indexed using the difference between post-treatment serum creatinine (RxCr) and baseline serum creatinine (BCr), or DeltaCr = RxCr - BCr) was the best predictor of graft survival time (r = 0.27, p < 0.0001) in patients experiencing AR. Subsequent AR episodes occurred more frequently in patients with a higher DeltaCr. However, DeltaCr was the best predictor of graft survival time (r = 0.44, p < 0.0001) in patients who had only one AR episode. The severity of the first AR episode (DeltaCr) correlated with the risk of subsequent AR episodes, the severity of a second AR episode and the average of mean blood pressures obtained after, but not before, the first AR episode. Therefore, we conclude that the severity of the first AR episode was the best predictor of renal allograft survival time in all patients experiencing AR, independent of subsequent AR episodes and correlated with subsequent clinical events which also influence renal allograft survival time. Thus, early diagnosis and aggressive treatment of the first AR episode are warranted to minimize AR severity and thereby maximize subsequent graft survival time. CONCLUSIONS: The severity of the first acute rejection episode following renal transplantation is the best determinant of graft survival time and correlates with subsequent clinical events which could further reduce graft survival time. PMID- 9850449 TI - Glipizide treatment of post-transplant diabetes does not interfere with cyclosporine pharmacokinetics in renal allograft recipients. AB - BACKGROUND: Glipizide is an oral antidiabetic drug that has been used in the treatment of post-transplant diabetes mellitus (PTDM). However, a published case report has indicated a possible interaction of glipizide with cyclosporine (CsA) pharmacokinetics in two renal transplant (tx) patients. The aim of this open prospective study was to investigate whether glipizide interacts with CsA pharmacokinetics in renal tx patients with PTDM. METHODS: Eleven renal tx patients (29-74 years of age) with PTDM who received Sandimmun Neoral as part of their immunosuppressive therapy were investigated. No patients had suffered any significant rise in serum creatinine (20%) from any cause over the last 2 wk before the study. Mean S-creatinine was 137 mumol/L (87-220). The mean CsA dose and whole blood concentration remained unchanged during the study. CsA whole blood concentrations were monitored over 12 h in all patients in random order, both on and off glipizide treatment. Blood samples were drawn immediately before the morning dose of CsA was given (trough) and 0.5, 1, 1.5, 2, 3, 4, 6 and 12 h after administration. RESULTS: Whole blood trough CsA concentration was not altered by glipizide treatment, 256 +/- 76 micrograms/L off and 242 +/- 73 micrograms/L (mean +/- SD) with glipizide coadministration. The area under the curve (AUC) and terminal half-life of CsA remained unchanged with glipizide treatment: 6391 +/- 1483 micrograms/L per h and 7.3 +/- 1.5 h without; and 6279 +/- 1601 micrograms/L per h and 7.1 +/- 1.8 h with glipizide, respectively. No change in the CsA peak concentration (Cmax) was observed: 1507 +/- 406 micrograms/L without and 1469 +/- 538 micrograms/L with glipizide coadministration. CONCLUSION: CsA pharmacokinetics is not significantly altered by glipizide coadministration. PMID- 9850450 TI - Limitations of pediatric donor kidneys for transplantation. AB - BACKGROUND: The shortage of grafts for kidney transplantation (Tx) has resulted in an expansion of criteria for pediatric donors. However, the limitations of pediatric donors for kidney Tx are not yet clearly established. We investigated the outcomes of recipients with pediatric kidney grafts from Colorado to elucidate the state of pediatric kidney grafts distributed in the nation and to clarify the limitations of pediatric donor kidneys for Tx. METHODS: Between January 1989 and July 1997, 675 organ donors in Colorado were utilized for Tx. Fifty-seven of these (8.4%) were 10 yr old or younger. Thirty-two of the 57 pediatric donors provided kidneys for 50 recipients both inside and outside Colorado. Forty-four of the 50 recipients were followed and are available for this study. RESULTS: Graft survival rates were 85.6, 77.3, 71.3 and 65.4% for 1, 2, 3 to 5, and 6 yr of follow-up, respectively. Thirteen recipients lost the grafts, including three graft losses within 2 wk, due to primary nonfunction, venous and arterial thrombosis. The mean serum creatinine value in 31 recipients with functioning grafts was 1.3 +/- 0.4 mg/dL at the time with follow-up of 45.9 +/- 28.3 months (5-96 months). Six grafts from 13-month to 3-yr-old donors were transplanted en-bloc and 5 of these have survived. Four of the 5 single grafts from 3 to 5-yr-old donors have functioned. Of the 33 grafts from donors aged 6-10 yr, including 30 single and 3 en-bloc grafts, 22 were still functioning. With respect to donor weight, 4 of the 5 en-bloc grafts from donors weighting 9-15 kg, and 6 of the 7 single and the 1 en-bloc grafts from donors weighing 15-20 kg were still functioning. Cold ischemic time (CIT) within 37.5 h was not associated with early graft outcome. CONCLUSIONS: The kidneys from donors 1-3 yr old and/or weighing 9-15 kg could be successfully transplanted en-bloc and those from donors more than 3 yr old and/or weighing 15 kg were best transplanted by single grafts. PMID- 9850451 TI - Serum total homocysteine concentration does not predict outcome in renal transplant recipients. AB - Established cardiovascular risk factors such as hypercholesterolemia have been claimed to adversely influence the outcome of renal transplants. The aim of the present study was to assess the effect of another risk factor, hyperhomocysteinemia, on graft outcome. This was relevant for two reasons; hyperhomocysteinemia is by now recognized as an independent risk factor for the development of atherosclerosis and it is highly prevalent in both dialysis patients and renal transplant recipients. The serum concentration of total homocysteine (tHcy) was analyzed in samples collected before transplantation in 81 patients and at 6 months after transplantation in 57 of these patients. Before transplantation, mean tHcy was 33.2 +/- 19.2 mumol/L and the prevalence of hyperhomocysteinemia was 94%. Six months after transplantation, mean tHcy was 27.7 +/- 14.6 mumol/L and the prevalence of hyperhomocysteinemia was 88%. The patients were followed up for 5 yr. Six months and 5 yr after transplantation, serum creatinine concentration and endogenous creatinine clearance were determined. After 6 months, allograft biopsy was evaluated. Neither pre- nor post transplant tHcy was found to influence patient or graft survival, graft function or histopathology. Thus, tHcy does not seem to predict either short-term or long term outcome of renal transplantation. PMID- 9850452 TI - Hyperacute rejection from antibody against class II HLA antigens. AB - The clinical significance of anti-B-cell antibodies in kidney and pancreas transplantation remains unresolved. Here, we report an isolated hyperacute rejection of the kidney, but not the pancreas, during a simultaneous kidney pancreas (SKP) transplant. The hyperacute rejection was due to IgG antibodies directed against class II antigens expressed on B-cells. Antibodies directed against class II antigens are generally not thought to produce hyperacute rejection, since class II antigens allegedly are minimally expressed on vascular endothelium in the kidney. The pancreas was spared and continues to function normally, suggesting that class II antigens were not strongly expressed in this pancreas. The differential susceptibility to B-cell antibodies of the two transplanted organs is noteworthy and should call attention to the danger from IgG antibodies to class II antigens in kidney transplantation. PMID- 9850453 TI - Necrosis of the femoral head after kidney transplantation. AB - We reviewed the medical records of 750 patients (445 men, 305 women), who had received a kidney transplant during the period 1968-1995, for any sign of necrosis of the femoral head. For post-operative immunosuppression, 374 patients had received high-dose corticosteroids (average 12.5 g during the first year post operatively), while 376 patients had received low-dose corticosteroids (average 6.5 g during the first year post-operatively) and cyclosporin A. Survival curves according to Kaplan and Meier (J Am Stat Ass 1958: 53: 457-481) were constructed. In the high-dose steroid group, 42/374 patients (11.2%) developed femoral head necrosis, at an average of 26.2 months post-transplantation. In the low-dose steroid group, only 19/376 (5.1%) patients developed this complication, at an average of 20.5 months post-transplantation. This difference in numbers of femoral head necroses was highly significant (p < 0.005). We conclude that steroid doses should be minimized whenever feasible in post-transplant immunosuppression therapy. PMID- 9850454 TI - Benefits of pre-emptive dose reduction for Sandimmune to Neoral conversion in stable renal transplant recipients. AB - In an effort to minimize nephrotoxicity resulting from greater exposure to cyclosporine after Sandimmune to Neoral conversion, we compared two conversion regimens using different dosing ratios. Serial serum creatinine concentrations and trough cyclosporine levels were measured in 26 patients converted from Sandimmune to Neoral using a 1:0.8 dosing ratio (Group 1) and compared to those of 26 patients converted using a 1:1 dosing ratio (Group 2). The percentage change in peak serum creatinine concentration after conversion was greater in Group 2. However, at last follow-up, the dose reductions in each group were comparable. Following conversion, patients in Group 1 required fewer dose adjustments and follow-up blood tests. Compared to conversion using a 1:1 dosing ratio, conversion from Sandimmune to Neoral using a 1:0.8 ratio results in comparable dose reductions and less short-term nephrotoxicity, while requiring less frequent laboratory monitoring. PMID- 9850455 TI - Management of recipients of hepatic allografts harvested from donors with malignancy diagnosed shortly after transplantation. AB - Transmission of undiagnosed malignancy with the graft is a dramatic complication of liver transplantation. Alternatives in the management of the recipients of livers, harvested from donors with malignancy diagnosed shortly after transplantation, are either early re-transplantation or close follow-up without re-operation. We reported 4 cases of liver recipients whose allografts were harvested from donors who were diagnosed with malignancy shortly after the liver transplantation. One recipient underwent re-transplantation, and the three other allografts were not removed. No recipient developed recurrence in the follow-up. While graft removal may be the only way to avoid tumor recurrence in recipients of liver graft harvested from donor with malignancy, close follow-up without re operation may also be considered. The risk of tumor transferral may depend on the histopathological aggressiveness and metastatic potential of the donor tumor, and may be low for low-grade, local tumors. This risk should be evaluated by analyzing large series, using databases of Eurotransplant or United Network for Organ Sharing. PMID- 9850456 TI - Successful pancreas and kidney transplantation: a view of metabolic control. AB - BACKGROUND: Pancreas and kidney transplantation (PKTx) has become the treatment of choice in uremic patients with insulin-dependent diabetes mellitus (IDDM). In these patients, the pancreatic graft provides an endogenous source of insulin. However, the determination of its long-term effect on metabolic control is crucial in order to establish the real efficacy of this treatment in diabetic patients. In this article, we describe the experience of the University of Barcelona in short- and long-term metabolic control in PKTx recipients. SUBJECTS AND METHODS: We performed seriated determinations of glycosilated hemoglobin (HbA1c) and oral glucose tolerance tests (OGTT) in 55 patients who underwent PKTx at our center from 1983 to 1996 with both grafts functioning for more than 1 yr (mean follow-up 4.2 +/- 2.3 yr). Basal glucose and insulin levels, areas under the curve (AUC) of glycemia and insulinemia after OGTT were analyzed throughout follow-up, as well as fasting insulin resistance index (FIRI). RESULTS: HbA1c levels throughout follow-up were within the normal range. The OGTT was normal in 80% of the patients 5 yr after transplantation. Hyperinsulinemia was present throughout follow-up. Over time, no differences were detected in the AUC of glycemia and insulinemia between controls and patients with PKTx. During the evolution, no differences were found in FIRI, in spite of increasing body weight. CONCLUSION: When successful, PKTx provides long-term normalization of glycemic control, assessed by HbA1c and OGTT, despite the existence of sustained hyperinsulinemia. Our results strongly suggest that PKTx is the most effective treatment for uremic patients with IDDM from a metabolic point of view. PMID- 9850457 TI - Immunosuppressive therapy with microemulsion cyclosporine A shortens the hospitalization of pediatric liver transplant recipients. AB - Intravenous administration of cyclosporine (Sandimmune) to rapidly and effectively achieve therapeutic serum levels in transplant recipients has been the treatment standard in many transplantation centers. With the development of microemulsion cyclosporine (Neoral), that standard is changing. Neoral has greater bioavailability than the oral form of Sandimmune and, consequently, can be more efficacious and cost-effective. To test this hypothesis, we undertook a retrospective study of Sandimmune and Neoral in the treatment of 66 children who underwent uncomplicated orthotopic liver transplantation in the Texas Medical Center between April 1991 and December 1997. Both forms of cyclosporine were evaluated in terms of in-patient treatment cost, recuperative time in the intensive care unit and duration of hospitalization. Twenty-two patients were treated orally with Neoral, and 44 patients were treated intravenously with Sandimmune for a mean time of 14 d. Once the blood concentration of Sandimmune reached a steady state, as confirmed by daily measurements of the trough level, the patients in the Sandimmune group were converted to oral cyclosporine. None of the 22 patients treated with Neoral required intravenous treatment. The mean time spent in the intensive care unit was 4 d for the Neoral group and 5.5 d for the Sandimmune group. The mean duration of hospitalization from the date of transplantation to discharge was 12 d for the Neoral group and 20 d for the Sandimmune group (p < 0.001). Based on these results, we determined that the overall cost per patient in the Neoral group was $3598 less than that per patient in the Sandimmune group. PMID- 9850459 TI - Cerebral mucormycosis after liver transplantation: a case report. AB - A fatal case of cerebral mucormycosis occurring shortly after liver transplantation is described. The patient was a 32-yr-old male with advanced end stage liver disease manifested by tense ascites, spontaneous bacterial peritonitis, deepening jaundice and anuria requiring hemodialysis. The 3rd day after successful liver transplantation the patient developed acute respiratory failure, then focal motor signs. Computed tomography showed fluid in the left maxillary sinus, partial opacification of the ethmoid and sphenoid sinuses, and diffuse low density lesions in both cerebral hemispheres. Despite treatment for cerebritis and cerebral edema, the patient's pupils became fixed and dilated, and brain death was declared. Autopsy revealed mucor sinusitis and cerebritis. Mucormycosis is an opportunistic fungal infection occurring in patients with diabetic ketoacidosis, malignancy, or immunodeficiency, and in those receiving wide-spectrum antibiotics, corticosteroids, or cytotoxic therapy. Mucor most frequently involves the face, rhinocerebral disease predominating. These infections are difficult to treat, but are curable with aggressive and frequent surgical debridement, discontinuation or reduction of immunosuppressive therapy and amphotericin. The diagnosis of mucormycosis is very difficult to make in cases such as the present one, in which the typical presentation and classical signs are not present. A high index of suspicion based on identified risk factors may assist in more rapid diagnosis of this life-threatening mycosis. PMID- 9850458 TI - Insulin allergy resolution following pancreas transplantation alone. AB - Clinically significant immunological reactions to exogenous insulin are classified as local or generalized. Most of the insulin allergies are local reactions which usually improve or resolve spontaneously. Generalized allergic reactions to insulin range in severity from simple urticaria to life-threatening anaphylaxis. Most of the allergic reactions to exogenous insulin are antibody mediated reactions to antigens such as zinc, protamine, non-insulin proteins, and aggregates of insulin molecules as well as animal antigens. Immunologic reactions to endogenous insulin usually result in insulin resistance. Herein, we report a case in which systemic insulin allergy was intractable, thus requiring a pancreas transplantation which is the first of its case according to the International Pancreas Transplant Registry/United Network for Organ Sharing (IPTR/UNOS) Registry. PMID- 9850460 TI - Thrombotic microangiopathy after liver-small bowel transplant. AB - We herein report the first case of immunosuppression-associated thrombotic microangiopathy (TMA) in which an extrarenal graft was primarily affected by the characteristic microvascular lesions. Although TMA is a well-known complication of cyclosporine (CSA) or tacrolimus therapy in renal and extrarenal (liver, heart, lung) transplant recipients, the kidney (transplanted or native) is typically the site primarily affected. We describe a combined liver-small bowel transplant recipient who developed tacrolimus-associated TMA that affected both her transplanted small bowel and her native kidneys. Involvement of the bowel, with evidence of microvascular occlusion on biopsy, led to the development of ischemic mucosal ulcers and eventual bowel perforation. Involvement of the kidney manifested with a doubling of the recipient's baseline serum creatinine level. Significant lowering of the tacrolimus dose resulted in healing of the small bowel ulcers and return to her baseline level of renal function. Therefore, it is important to note that, in transplant recipients, TMA with microvascular occlusion may affect extrarenal sites. In small bowel transplant recipients, the result might be ischemic ulcers in the graft and eventual bowel perforation. PMID- 9850461 TI - Lipid modification strategies in the production of nutritionally functional fats and oils. AB - Rapid improvements in the understanding of the nutritional requirements of both infants and adults has led to new developments in the modification of fats and oils. Specific targets include the improvement in growth and development of infants, treatment of disease in adults, and disease prevention. Efforts have been focussed on the production of structured lipids using medium-chain acids and long-chain polyunsaturated fatty acids (PUFAs), as well as the concentration of long-chain PUFAs from new and existing sources. Short- and medium-chain fatty acids are metabolized differently than long-chain fatty acids and have been used as a source of rapid energy for preterm infants and patients with fat malabsorption-related diseases. Long-chain PUFAs, specifically docosahexaenoic acid and arachidonic acid, are important both in the growth and development of infants, while n-3 PUFAs have been associated with reduced risk of cardiovascular disease in adults. Based on the requirements for individual fat components by different segments of the population, including infants, adults, and patients, ideal fats can be formulated to meet their needs. By using specific novel fat sources and lipid modification techniques, the concentrations of medium-chain, long-chain saturated, and long-chain polyunsaturated fatty acids as well as cholesterol can be varied to meet the individual needs of each of these groups. While genetic modification of oilseeds and other novel sources of specific lipid components are still being developed, chemical and lipase-catalyzed interesterification reactions have moved to the forefront of lipid modification technology. Fractionation of fats and oils to provide fractions with different nutritional properties has potential, but little work has been performed on the nutritional applications of this method. The choice of suitable lipid modification technologies will depend on the target lipid structure, production costs, and consumer demand. A combination of some or all of the present lipid modification techniques may be required for this purpose. PMID- 9850462 TI - A critical review of the quality and safety of BADGE-based epoxy coatings for cans: implications for legislation on epoxy coatings for food contact. AB - BADGE-based epoxy resins have been commercially available for more than 40 years. They are extremely versatile, finding applications in many fields in both thermal and ambient cure applications. The present review focuses on their use in the food industry as surface coatings for food and beverage cans, and in particular on their chemical stability in relation to their toxicology and sensorial effects on foods. The implications of these properties of epoxy resins for prospective EU legislation on plastic coatings for food contact is discussed. PMID- 9850463 TI - Structure and technofunctional properties of protein-polysaccharide complexes: a review. AB - Food proteins and polysaccharides are the two key structural entities in food materials. Generally, interactions between proteins and polysaccharides in aqueous media can lead to one- or two-phase systems, the latter being generally observed. In some cases of protein-polysaccharide net attraction, mainly mediated through electrostatic interactions, complex coacervation or associative phase separation occurs, giving rise to the formation of protein-polysaccharide complexes. Physicochemical factors such as pH, ionic strength, ratio of protein to polysaccharide, polysaccharide and protein charge, and molecular weight affect the formation and stability of such complexes. Additionally, the temperature and mechanical factors (pressure, shearing rate, and time) have an influence on phase separation and time stability of the system. The protein-polysaccharide complexes exhibit better functional properties than that of the proteins and polysaccharides alone. This improvement could be attributed to the simultaneous presence of the two biopolymers, as well as the structure of the complexes. Consequently, the interesting hydration (solubility, viscosity), structuration (aggregation, gelation) and surface (foaming, emulsifying) properties of these complexes can be used in a number of domains. Among others, these could be macromolecular purification, microencapsulation, food formulation (fat replacers, texturing agents), and synthesis of biomaterials (edible films, artificial grafts). PMID- 9850464 TI - Demography and the environment. AB - Demographers' interest in the environment has generally been enmeshed in broader issues of population growth and economic development. Empirical research by demographers on environmental issues other than natural-resource constraints is limited. In this paper, I briefly review past demographic thinking about population and the environment and suggest reasons for the limited scope of demographic research in this area. Next, I describe more recent demographic research on the environment and suggest several newer areas for demographic research. Finally, I consider the future of research on the environment in the field of demography. PMID- 9850465 TI - Deceleration in the age pattern of mortality at older ages. AB - The rate of mortality increase with age tends to slow down at very old ages. One explanation proposed for this deceleration is the selective survival of healthier individuals to older ages. Data on mortality in Sweden and Japan are generally compatible with three predictions of this hypothesis: (1) decelerations for most major causes of death; (2) decelerations starting at younger ages for more "selective" causes; and (3) a shift of the deceleration to older ages with declining levels of mortality. A parametric model employed to illustrate the third prediction relies on the distinction between senescent and background mortality. This dichotomy, though simplistic, helps to explain the observed timing of the deceleration. PMID- 9850467 TI - Contraceptive use in South Africa under apartheid. AB - In this paper, patterns of contraceptive use among black South African women in the late 1980s are examined. Multilevel logit models are used to evaluate the extent to which segregation of the African population into homelands gave rise to uneven patterns of contraceptive use; how this pattern was shaped by variations in family-planning acceptability; and the way in which the system of male labor migration and social and economic inequities across communities affected women's use of contraceptives. Results show that variation in contraceptive use across homeland areas diminished with the addition of community controls for development and migration. Controlling for unobserved heterogeneity indicated that localized conditions could offset the advantages or disadvantages of living in a former homeland. PMID- 9850466 TI - A methodology for identifying married couples in Medicare data: mortality, morbidity, and health care use among the married elderly. AB - We describe a method for the development of cohorts of up to three quarters of the 14 million married couples aged 65 and over in the United States. The health care experiences, illness histories, and mortality of these identified couples can be assessed longitudinally using Medicare data. We summarize strengths and limitations of using data from Medicare administrative records for the study of marriage, health, and aging. We illustrate the method by demonstrating substantial differences in survival in a cohort of hospice patients as a function of not only the patient's own diagnosis and illness burden but also the patient's spouse's illness burden. PMID- 9850469 TI - Three strikes and you're out: demographic analysis of mandatory prison sentencing. AB - Much of the debate about the costs and benefits of "three-strikes" laws for repeat felony offenders is implicitly demographic, relying on unexamined assumptions about prison population dynamics. However, even state-of-the-art analysis has omitted important demographic details. We construct a multistate life-table model of population flows to and from prisons, incorporating age specific transition rates estimated from administrative data from Florida. We use the multistate life-table model to investigate patterns of prison population growth and aging under many variants of three-strikes laws. Our analysis allows us to quantify these demographic changes and suggests that the aging of prison populations under three-strikes policies will significantly undermine their long run effectiveness. PMID- 9850468 TI - The long-term impact of parents' childbearing decisions on children's self esteem. AB - We examine the long-term impact of parents' childbearing decisions on children's self-esteem. We focus on subjective aspects of the home environment in the creation of children's internalized sense of self-worth. Unique 23-year family panel data combining measures of mothers' childbearing, mothers' childbearing intentions, and children's self-esteem allow us to examine the overall links between parents' childbearing and children's self-esteem. The results demonstrate that parents' childbearing intentions can have a significant long-term impact on their children's self-esteem. Children who were unintended by their mothers have significantly lower self-esteem 23 years later. Our findings indicate that giving birth to an unintended child can have a long-term negative impact on subjective aspects of the child's well-being, at least in terms of self-esteem. Unintended childbearing has received an increasing amount of research attention in recent years. PMID- 9850470 TI - Does it matter who answers the race question? Racial classification and income inequality in Brazil. AB - Previous studies of racial inequality have relied on official statistics that presumably use self-classification of race. Using novel data from a 1995 national survey in Brazil, we find that the estimates of racial income inequality based on self-classification are lower than those based on interviewer classification. After human capital and labor market controls, whites earn 26% more than browns with interviewer classification but earn only 17% more than browns with self classification. Black-brown differences hardly change: Blacks earn 13% and 12% less than browns with interviewer classification and self-classification, respectively. We contend that interviewer classification of race is more appropriate because analysts of racial inequality are interested in the effects of racial discrimination, which depends on how others classify one's race. PMID- 9850471 TI - Revisiting occupational sex segregation in the United States, 1910-1990: results from a log-linear approach. AB - I reexamine trends in the strength and structure of occupational sex segregation in the United States from 1910 to 1990. Log-multiplicative models show significant change in the association between gender and occupation. Contrary to conventional characterizations, a substantial proportion of this change occurred before 1970. Likewise, a margin-free index shows more integration over the century than do conventional indices. These discrepancies arise from occupation specific variations in the trajectory of sex segregation: Highly segregated occupations were especially likely to integrate between 1930 and 1940. I identify regions of the occupational structure and pivotal periods in which shifts in segregation occurred and compare these results with conventional historical accounts. PMID- 9850472 TI - Occupational gender segregation: index measurement and econometric modeling. AB - Empirical studies of gender segregation by occupation must be founded on rigorous measurement procedures. There appears to be a consensus that any index used in the analysis of time-series or international cross-section employment data must be either margin-free or decomposable to yield a margin-free component. On the other hand, Charles and Grusky (1995) advocate the use of multiplicative log models from which a margin-free odds ratio can be derived. In this paper, I contrast the construction and interpretation of the index of dissimilarity and the Karmel-MacLachlan index with the multiplicative modeling of gender segregation and the associated log index. PMID- 9850473 TI - The past, present, and future of sex segregation methodology. AB - We review the logic underlying margin-free analyses of sex segregation arrays. In the course of our review, we show that the Karmel-MacLachlan decomposition does not live up to its margin-free billing, as the index upon which it rests, Ip, is itself margin-sensitive. Moreover, because the implicit individualism of D is necessarily inconsistent with margin-free analysis, the field would do well to abandon not merely the Karmel-MacLachlan decomposition but all related efforts to purge marginal dependencies from D-inspired measures. The criticisms that Watts (1998) levels against our log-multiplicative approach are likewise unconvincing. We demonstrate that our preferred models pass the test of organizational equivalence, that the "problem" of zero cells can be solved by applying well developed methods for ransacking incomplete or sparse tables, and that simple log multiplicative models can be readily devised to analyze disaggregate arrays. We illustrate these conclusions by analyzing a new cross-national archive of detailed segregation data. PMID- 9850474 TI - The analysis of sex segregation: when is index measurement not index measurement? AB - In their paper in this issue, Grusky and Charles (1998) make a number of dubious claims about the measurement and interpretation of sex segregation. First, they incorrectly claim that only log-odds measures yield margin-free measures of segregation. Second, the estimation and testing of a limited class of log-linear models does not provide an independent test of the appropriateness of a log-odds ratio index to measure segregation. Their estimation in forms them of the statistically justifiable degree of occupational disaggregation, not whether a log-odds ratio is superior to, say, a linear index in the measurement of segregation. Finally, their index A is beset with problems of interpretation, not withstanding their arguments, and their additional measures, AW and AB, suffer similar problems. Grusky and Charles are, however, correct in arguing that measurement procedures should be margin-free. Further, I concur with the view that the adoption of a single annual summary measure of segregation cannot be justified, because it is premised on the assumption that individual occupations, or groups of occupations, exhibit similar trends in sex segregation. PMID- 9850475 TI - Comment on "Compromised birth outcomes and infant mortality among racial and ethnic groups". AB - Frisbie, Forbes, and Pullum (1996) show that it is meaningful to account for low birth weight, preterm delivery, and intrauterine growth-retardation when analyzing differences in compromised birth outcomes and infant mortality among racial and ethnic groups. I compare their findings for the 1987 U.S. birth cohort with findings for the 1988 U.S. birth cohort, using linked birth and infant death vital statistics from the National Center for Health Statistics. I focus on their calculation of fetal growth curves, which are highly at odds with the curves commonly used in the obstetric and pediatric literature. I compare birth outcome distributions and infant death probabilities using Frisbie et al.'s method and other standards. I conclude that Frisbie et al.'s method is not suited for the study of intrauterine growth-retardation at the population level because of the major flaws in gestational age measurement that exist in the type of data they use. An appropriate alternative is to apply a standard of normal intrauterine growth derived from antenatal estimation of fetal weight-for-gestational-age to the vital statistics data. PMID- 9850477 TI - The Worldwide Burden of Diabetes. Proceedings of a workshop. Phoenix, Arizona, USA. 6-7 December 1996. PMID- 9850476 TI - Birth outcome, not pregnancy process: reply to van der Veen. AB - In a recent article (Frisbie, Forbes, and Pullum 1996) we documented racial/ethnic differences in birth outcomes according to a more fine-grained classification than has typically been employed in the demographic literature. In his commentary, van der Veen focuses on the measurement of one of the dimensions of that classification, maturity of the infant, as proxied by the fetal growth ratio. The crux of the critique is easily seen in van der Veen's statement that "all of my disagreements with Frisbie et al.'s method arise from their particular use of a postnatal standard for the assessment of intrauterine growth." Our critic misunderstands our objective: He fails to realize our interest in birth outcome, not pregnancy process, and does not perceive that our intent was to extend the research extant in both the demographic and public-health literatures in which patently postnatal (i.e., ex utero) measures are taken as outcomes interesting in their own right and/or as risk factors for infant mortality and infant and childhood morbidity. Specifically, he does recognize that we purposefully expanded our focus to include moderately compromised births to determine if they were at higher risk than the normal births with whom they are conventionally categorized. Our discussion draws on research cited in the original article, on studies cited by our critic, and on a few more recent investigations. Although we have never argued that ours is the only, or even the best approach in all cases, we try to clarify the rationale for, and adduce additional empirical evidence of, the utility of the method we used. PMID- 9850478 TI - Epidemiology of type 2 diabetes: risk factors. AB - A number of cross-sectional and prospective studies that compared the insulin sensitivity of various national and ethnic populations within the U.S. to the total U.S. population were analyzed to find possible risk factors for the development of type 2 diabetes. It was found that the risks for diabetes in African-Americans, Hispanics, and Native Americans are approximately 2, 2.5, and 5 times greater, respectively, than in Caucasians. Studies of the prevalence of type 2 diabetes in Mexican Americans and non-Hispanic whites in San Antonio showed that there is an inverse relationship between socioeconomic status and the prevalence of diabetes. It also appears that cultural effects lead to an increased incidence of obesity in these populations, which may lead to insulin resistance. Genetic factors may also be a contributing factor. A 5-year, prospective study of insulin resistance in Pima Indians showed a relationship between impaired glucose tolerance and subsequent development of type 2 diabetes. In a 7-year study in Mexican Americans, those subjects who had both high insulin secretion and impaired insulin sensitivity had a 14-fold increased risk of developing type 2 diabetes. Regardless of cultural and ethnic factors, the San Antonio Heart Study, which compared Mexican Americans and non-Hispanic whites, showed that in both groups, the strongest predictors of developing type 2 diabetes are elevated fasting insulin concentrations and low insulin secretion. PMID- 9850479 TI - The economic impact of diabetes. AB - In assessing the economic impact of diabetes for a population, several factors should be considered, including the incidence and prevalence of the disease, the level of development of the health care system, and the population's overall level of economic development. Two different approaches have been used to address the economic impact of an increasing incidence of diabetes. The first approach uses disability-adjusted life-years (DALYs) to measure intangible costs associated with diabetes. It combines the number of healthy life-years lost as a result of early mortality with those lost because of disability. The second approach, which has been used more frequently, is the cost-of-illness approach, which includes the concepts of direct, indirect, and intangible costs. A study conducted by the World Bank found that of the 1,362 million DALYs lost to all illnesses in 1990, 7.97 million DALYs were lost because of diabetes. In a 1992 study that assessed the direct costs of treating diabetes in the U.S., the American Diabetes Association used the cost-of-illness approach and found that the estimated total expenditure for 1 year was $45.2 billion. The 1994 epidemiological studies by Zimmet and the World Health Organization include estimates of increased prevalence of diabetes resulting from an increase in population. Estimates of the global cost of diabetes based on these studies reveal that diabetes accounts for 2-3% of the total health care budget in every country; therefore, an increase in diabetes incidence and prevalence translates into a significant economic impact. PMID- 9850480 TI - Diabetes in America: epidemiology and scope of the problem. AB - Epidemiological studies performed over the past 40 years have shown that the prevalence of diagnosed diabetes has increased dramatically in the U.S. and that a substantial proportion of the population has undiagnosed diabetes, impaired fasting glucose, and impaired glucose tolerance. Diabetes is most prevalent in minority populations, such as African-Americans, Native Americans, and Mexican Americans. Increasing prevalence of diabetes has led to increases in microvascular complications such as blindness, end-stage renal disease, and lower limb amputations. Poor glycemic control contributes to the high incidence of these complications, yet community-based studies of diabetic patients show their mean fasting plasma glucose concentration is generally > 180 mg/dl compared with 100 mg/dl for nondiabetic individuals. In people with diabetes, risk factors for cardiovascular disease including elevated fasting plasma glucose, blood pressure, total cholesterol, triglycerides, and obesity partly explain the high proportion of deaths (60-70%) caused by cardiovascular disease in people with diabetes. More intensive diabetes management and improved glycemic control could minimize long term complications of the disease and would be expected to reduce the morbidity, mortality, and costs associated with diabetes. PMID- 9850481 TI - The public health burden of diabetes and the reality of limits. AB - Improvements in diabetes surveillance, diagnosis, and treatment have, in recent years, heightened awareness of the burden of diabetes and aroused concern about the amount of health care resources that will be necessary to manage this disease effectively in the future. Examination of diabetes from the twin perspectives of economics and public health challenges basic notions of the health care tradition in the Western world: the real-world combination of finite resources and the growing need/demand for health services forces the consideration of limits in the provision of health care. The growing need to rationally allocate limited health care resources poses emotional, potentially divisive questions of science, politics, economics, and ethics that patients and physicians must each address. PMID- 9850483 TI - Diabetes and the promise of managed care. AB - The term managed care has a positive connotation to many people and a negative connotation to others. It implies a systematic approach to the provision of care supported by a managed care organization, the underlying values of which are efficiency in care delivery and continuous health improvement of the population being served. The recent advances in knowledge surrounding the treatment of diabetes, the codification of these advances into treatment guidelines, and the significant gap between recommended and real-world treatment patterns all suggest that there is an opportunity for managed care organizations to play a beneficial role in improving the treatment of people with diabetes. Typical strategies used by managed care organizations include dissemination of treatment guidelines and information feedback to providers and patients to suggest diagnostic measures or therapeutic interventions that should be repeated with regularity to maintain or improve the health of people with diabetes. Improvements in population-level measurement of health care quality will increase the focus on preventable long term complications of diabetes and other chronic diseases and may establish increasingly quantitative quality metrics that should improve real-world treatment patterns. Models exist for a multifaceted approach to improve the care of people with diabetes through such mechanisms; this article reviews the experience of one health plan with such undertakings. PMID- 9850482 TI - The effects of treatment on the direct costs of diabetes. AB - Treatment of diabetic complications consumes health care resources. Intensive therapy was shown by the Diabetes Control and Complications Trial (DCCT) to avert complications. Economic analyses and models have been used to evaluate the cost effectiveness of intensive therapy for people with type 1 and type 2 diabetes. An economic analysis of the DCCT estimated the cost of intensive therapy to be two to three times greater than that of conventional therapy. In contrast, an economic model predicts that intensive therapy, as compared with conventional therapy, could reduce blindness from 34 to 20% or by 41%, end-stage renal disease from 24 to 7% or by 71%, and lower-extremity amputations from 7 to 4% or by 43%. Although intensive therapy is more expensive, when the costs of complications are factored in, it becomes cost-effective for treatment of type 1 diabetes. Similarly, a model to evaluate the cost-effectiveness of intensive therapy for people with type 2 diabetes found that the lifetime costs of general and diabetes related medical care would be approximately two times greater. However, the reduction in lifetime costs of complications, which would produce substantial reductions in costs of treatment, largely offsets the difference. Intensive therapy for type 1 and type 2 diabetes may be more expensive than conventional therapy, but from an economic perspective, it is comparable in cost to pharmacological therapies for people with hypertension and hypercholesterolemia. From a health system viewpoint, intensive therapy represents a fruitful long-term financial investment. PMID- 9850484 TI - The burden of diabetes: the patient's perspective. PMID- 9850485 TI - Reducing the burden of diabetes. The National Diabetes Education Program. AB - The National Diabetes Education Program (NDEP) is a joint initiative sponsored by the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health and the Division of Diabetes Translation of the Centers for Disease Control and Prevention. The program will develop a partnership of public and private organizations and implement educational activities to help reduce the morbidity and mortality associated with diabetes and its complications. Participating public and private organizations include the Indian Health Service, the Department of Veterans Affairs, the American Diabetes Association, the American College of Physicians, the American Dietetic Association, and the American Association of Diabetes Educators. Key program activities will include (1) ongoing public awareness and education (2) and identification and dissemination of existing educational tools and resources for patients and their families, health care providers, and payers and purchasers of health care. The NDEP will also sponsor the development and dissemination of new educational tools and resources, including science-based guidelines, efforts to improve quality and access to diabetes care, and program evaluation. PMID- 9850487 TI - The U.K. Prospective Diabetes Study. A review. AB - The objective of the U.K. Prospective Diabetes Study (UKPDS), initiated in 1977, was set up to determine whether improved blood glucose control in people with type 2 diabetes will prevent the complications of diabetes. The UKPDS was also designed to determine whether there are differences between conventional policy (diet therapy) and three different regimens of intensive treatment policy, based on sulfonylurea, metformin, or insulin. Interim efficacy analyses revealed that the intensive policies with sulfonylurea, insulin, and metformin were equally effective in reducing fasting plasma glucose concentrations. However, glucose and HbA1c measurements steadily increased with time, reflecting ongoing deterioration of beta-cell function. Cardiovascular disease was the major cause of complications, and the risk factors included raised LDL cholesterol concentrations, low HDL cholesterol concentrations, elevated blood pressure, elevated HbA1c concentrations, and smoking. A final study report was issued in September 1998, when the median duration of therapy was 11 years. PMID- 9850486 TI - The burden of chronic hyperglycemia. AB - Diabetic complications constitute the principal clinical and economic burdens of diabetes. Available evidence indicates that sustained reduction in hyperglycemia (as measured by HbA1c) reduces the incidence of diabetic complications, including microvascular and neurological disease, and will likely reduce the risk of macrovascular disease. In the U.S., targeting interventions to aged and ethnic subpopulations, which are currently receiving suboptimal care, and increasing efforts to lower the average HbA1c concentrations one or two percentage points in people with diabetes would be a more cost-effective goal than aiming for a theoretical HbA1c threshold below which the risk of complications is negligible. Other risk factors for diabetic complications, such as obesity, hypertension, cigarette smoking, and hypercholesterolemia, must also be addressed to reduce the burden of diabetes. PMID- 9850488 TI - Relation of glycemic control to diabetic complications and health outcomes. AB - Data from the Wisconsin Epidemiologic Study of Diabetic Retinopathy (WESDR) demonstrated that the incidence of diabetic complications is directly related to glycemic control. The results of the Diabetes Control and Complications Trial and Stockholm Study showed that intensive insulin therapy and improved glycemic control reduced diabetic complications in people with type 1 diabetes. Results of the U.K. Prospective Diabetes Study Group and the Kumamoto trial also support the relationship between glycemic control and diabetic complications in individuals with type 2 diabetes. Preliminary WESDR health outcomes data suggest that higher levels of glycemia are related to a decreasing quality of life. This study and others showing that higher levels of glycemia are associated with an increased incidence of complications suggest that it is the complications of diabetes that contribute to a decrease in quality of life. Despite evidence of the benefits of improved glycemic control, a large percentage of people with diabetes maintain poor glucose control in part because of the limitations of the therapies available for diabetes management. PMID- 9850490 TI - HIV/AIDS: a South African perspective. PMID- 9850489 TI - Valuing quality of life and improvements in glycemic control in people with type 2 diabetes. AB - Outcomes research is used increasingly for assessing the health economic benefits of new therapeutic programs and interventions. The measurement properties of the outcomes assessment tools are important. If overlooked, they can mislead health care administrators and caregivers regarding the importance and value of these programs and interventions. We reviewed the literature and conducted two analyses to determine the absolute, relative, and operative quality-of-life ranges for people with type 2 diabetes. Quality of life and fasting blood glucose and HbA1c concentrations were measured at baseline and at 4, 8, and 12 weeks of treatment in 569 men and women randomized to either glipizide gastrointestinal therapeutic system (GITS) or placebo in a double-blind, multicenter clinical trial. A subgroup of 290 patients completed a diabetes-specific health states questionnaire at endpoint (week 12 or early termination) rating 10 health-state descriptions on a health thermometer scale ranging from 0 (death) to 100 (full health). Health losses at the higher end of the scale had a greater negative utility than did comparable losses at lower health states, indicating patients' strong preferences for maintaining asymptomatic or mildly symptomatic conditions. Patients rated their current health state at 83.4 +/- 0.8% of full health and indicated that a loss of 27 points below this value would prevent them from living and working as they currently do. The calibration analysis applied to the quality-of-life scales suggested that the targeted range for clinical investigation and quality-of-care evaluation must be more narrowly focused. Effect sizes as seemingly small as 2% (0.25 responsiveness units) on the absolute scale can correspond to quality-of-life losses of 15-20% on the personal operative scale. Differences in glycemic control clearly affected quality of life. Those patients with the best HbA1c responses (decreasing 1.5% or more from baseline) versus those with the worst responses (increasing 1.5% or more from baseline) were separated by 0.6 responsiveness units for the overall quality-of life summary measure. The calibration analysis suggested that this degree of better glycemic control provides a nearly 50% gain in quality of life according to personal expectations within the operative range. In conclusion, general measures of quality of life may be too crude and insensitive to capture the important gains in health outcomes due to new therapeutic interventions and programs in diabetes. Quality-of-care evaluations for diabetes are at risk of favoring inferior programs with lower costs simply because gains or losses in health outcomes go undetected. PMID- 9850491 TI - The ethical issues involved in organ transplant. PMID- 9850492 TI - Brazilian nurses play pivotal role in organ transplant. PMID- 9850493 TI - Bringing the World together to fight HIV/AIDS. AB - There were some significant first for nursing at the 12th World AIDS Conference in Geneva: a special nursing session, the concept of "care" in one of the four tracks and ICN as a partner in organizing the event. The major drive was "bridging the gap" between the have and have not in access to therapies and continuum of care. At a Pre-Conference Nursing Satellite Meeting nurses exchanged ideas and research and were provided another perspective of the HIV/AIDS epidemic -the continually widening gap between the North and South--to which nurses should respond in unison. PMID- 9850494 TI - Nurses are the key to effective HIV/AIDS prevention and care. AB - "Connecting Experience and Knowledge"--the theme of a recent conference organized by the European Association of Nurses in AIDS Care (EANAC)--is at the heart of effective HIV/AIDS care, noted ICN President Kirsten Stallknecht in her opening statement. Below, excerpts from her speech. PMID- 9850495 TI - Global overview of HIV/AIDS--a challenge for nurses. AB - The human immunodeficiency virus (HIV) continues to spread around the world, insinuating itself into communities previously little troubled by the epidemic and strengthening its grip on areas where AIDS is already the leading cause of death in adults. (See global estimates on page 176.) Below, the evolving picture region by region. PMID- 9850496 TI - The HIV/AIDS epidemic in Malawi. AB - In Africa south of the Sahara desert, nearly 21 million people are living with HIV/AIDS. In 13 countries in the region at least 10% of all adults are infected with HIV/AIDS and the prevalence rates in many capital cities are 35% or more. Botswana and Zimbabwe have reached a prevalence rate of 25%, or one out of every four adults in the country is infected. And in nearby Malawi, the prevalence rate is 15% among adults and increasing daily. The Malawi Government recognizes HIV/AIDS as a serious public health and socioeconomic problem requiring multisectoral involvement. Below is an overview of the epidemic's magnitude, the cultural practices facilitating its proliferation and how nurses are joining the country's community mobilization strategy. PMID- 9850497 TI - Teaching about HIV/AIDS: the lessons learned. AB - To learn and teach about HIV/AIDS is to enter complex and sensitive territory that at times may be personally challenging. In the vast literature on HIV infection and nursing two main themes recur: nurses' lack of understanding, negative attitudes and anxieties related to HIV/AIDS and the need for education to change such attitudes and enhance nurses' knowledge and skills. PMID- 9850498 TI - Four ways to teach PHC in a changing world. AB - Nurse educators are challenged to revise teaching/learning methods to support community-based primary health care (PHC) systems. The key elements are: integrating education with the realities of practice and people's needs; moving communities to strive towards solving their own health problems; and recognizing the socioeconomic health determinants. Described below are four educational methods based on PHC: inquiry-based learning, IMPACTS (a game to encourage multisectoral collaboration), distance education and community-based education. PMID- 9850499 TI - Connecting to Internet--search tools. AB - The amount of information on the Internet is growing by the nanosecond. It is no longer expeditious to "surf" site to site now that automated convenient-to-use search tools can find the information you are looking for swiftly. Unfortunately, no one tool covers the entire worldwide web (www), much less the entire Internet. The following describes the components and characteristics of popular Internet search tools. PMID- 9850500 TI - Direct stimulatory effects of insulin-like growth factor-I (IGF-I) on nuclear RNA polymerase II activity and overall protein synthesis in immature rat Sertoli cells. AB - A large body of evidence support the existence of an intratesticular Insulin-like Growth Factor I (IGF-I) system that can be viewed as a positive regulator of testicular functions. IGF-I may act at the testis level as a paracrine and autocrine differentiating factor. In the present study the role of IGF-I on Sertoli cell protein synthesis at transcriptional level has been investigated by evaluating the effect of IGF-I on nuclear RNA polymerase II activity as well as on total protein synthesis. Sertoli cells isolated from midpubertal rats and cultured in the presence of physiological doses of IGF-I showed a significant increase in nuclear RNA polymerase II activity (+80%) which appears to be correlated with a 50% increase in overall protein synthesis and a 40% increase in Androgen Binding Protein (ABP) production. These data provide the first evidence for a conceivable role of IGF-I in the modulation of Sertoli cell development through a direct action at the transcriptional level resulting in augmented protein synthesis. PMID- 9850501 TI - Respiratory activity and growth of human skin derma fibroblasts. AB - A study has been made on the speed of growth and respiratory activity of fibroblast cultures from control derma, cheloid (hypertrophic) scar and stabilized scar taken from human skin. The speed of growth and the efficiency of plaque formation of fibroblasts from cheloid scar were greater in comparison with those of fibroblasts from stabilized scar and were stimulated by the addition to the culture medium of the exudate from post-traumatic ulcer. Measurement of the contents of cytochromes showed a decrease in the content of cytochromes b562 and c + c1 in the fibroblast culture from both cheloid and stabilized scar as compared to the fibroblast culture from control derma. Cytochrome aa3 content did not show significant difference among the three types of fibroblast cultures. The respiratory activities supported by pyruvate plus malate, succinate or ascorbate plus N,N,N',N'-tetramethyl-p-phenylenediamine did not show, however, significant difference among the three fibroblast cultures. These observations show that the speed of growth of skin fibroblasts does not depend on the overall respiratory capacity. The exudate stimulated the activity of cytochrome c oxidase in fibroblasts from control derma, and cheloid scar. This effect and the accompanying stimulation of fibroblast growth might be correlated with the balance of oxygen free radicals. PMID- 9850502 TI - Mobius' syndrome: features and etiology. AB - PURPOSE: To review the clinical features and possible aetiology of all cases of Mobius' syndrome presented at the Royal Alexandra Hospital for Children in Sydney, Australia, from 1963 through 1995. METHODS: All charts of patients suffering from Mobius' syndrome were reviewed, and each patient was thoroughly assessed by a geneticist and a pediatric ophthalmologist. RESULTS: Patterns of systemic and ocular clinical features became evident. Of 23 patients in the series, 10 (43%) had a history significant events in utero. CONCLUSIONS: All patients suffering from Mobius' syndrome require thorough assessment by a multidisciplinary team including a geneticist or pediatrician and a pediatric ophthalmologist. Significant vascular events of pregnancy may explain some cases of fetal vascular disruption to the brainstem. PMID- 9850503 TI - Assessing visual function in children younger than 1 1/2 years with normal and subnormal vision: evaluation of methods. AB - PURPOSE: Methods for detecting subnormal vision early in life are needed to adequately diagnose the condition and begin treatment and rehabilitation. METHODS: Forty-six children ages 1 month to 1 1/2 years with either assumed normal vision, visual impairment due to eye disease, or strabismus were examined with the following tests: Stycar rolling balls; preferential looking (Teller acuity cards); and the ability to detect raisins, puffed rice, and sugar strands on two different backgrounds, one white and one black. RESULTS: Children with visual impairment and strabismus often showed normal values for their age, and children with assumed normal vision sometimes had values below those established for normal development of vision with the Stycar rolling balls and the Teller acuity cards. CONCLUSIONS: It is not possible to use these tests to reliably differentiate among normal vision, visual impairment, or strabismus in children younger than 1 1/2. Not until recognition acuity, symbols, or letters can be tested can reliable results be obtained. However, the tests are useful to estimate the general visual function of children in this age group. PMID- 9850504 TI - Ultrasound biomicroscopy of the anterior segment of the eyes of infants. AB - BACKGROUND: Our objective was to evaluate the new method of ultrasound biomicroscopy in examining the anterior segment of the eyes of infants. METHODS: The infant was held supine during examination, which was performed under topical anesthesia only. One drop of proparacaine was placed in the eye, and a small lid speculum was inserted. The palpebral fissure was filled with 2.5% methylcellulose as a coupling medium. The ocular cup was not used. Scanning was performed by placing the probe close to the area of interest. RESULTS: Case 1: A 3230-g male infant was delivered by cesarean section at 39 weeks' gestation. The left eye was studied with ultrasound biomicroscopy. Examination revealed a narrow angle with the anterior chamber angle of 25 degrees. The trabecular-ciliary process distance was 510 microns. The depth of the anterior chamber was 1414 microns. Case 2: A 1368-g female infant was delivered by cesarean section at 30 weeks' gestation. The left eye was studied. The anterior chamber angle was 16 degrees. The trabecular-ciliary process distance was 380 microns. The depth of the anterior chamber was 1134 microns. CONCLUSION: Ultrasound biomicroscopy is clinically feasible in infants and has a potential for use in clinical research. Additional study is needed to assess the reproducibility of the measurements and to acquire additional clinical experiences. PMID- 9850505 TI - Sarcoidosis presenting as multilobular limbal corneal nodules. AB - PURPOSE: To review reported external ocular manifestations of sarcoidosis and to present bilateral, multilobular, nodular, limbal, corneal nodules as being a unique manifestation of the disease. PATIENTS AND METHODS: A 16-year-old Saudi girl presented with bilateral, multilobular, solid, limbal nodules, with a vascular supply from the conjunctival vessel, and associated membraneous conjunctivitis and healed trachoma. The Schirmer's test revealed less than 2 mm in both eyes with tear meniscus less than 2 mm. Biopsy of an associated palpebral conjunctival nodule was performed, in addition to a gallium scan, chest X-ray, and a serum angiotensin-converting enzyme (SACE) level. RESULTS: The culture showed beta-hemolytic streptococci. Gallium scan showed intake by both lacrimal glands. Her chest X-ray results were normal, as was the SACE level. Biopsy of the excised conjunctival nodule disclosed a noncaseating granulomatous reaction with epithelioid and giant cells, and chronic inflammatory cell infiltrate confirming a diagnosis of sarcoidosis. CONCLUSION: A multilobular, nodular, perilimbal mass as a unique manifestation of sarcoidosis is presented. A streptococcal membraneous conjunctivitis and healed trachoma superimposed. A review of sarcoidosis of the external eye is included. PMID- 9850506 TI - A ring hypopyon in a patient with meningococcal endophthalmitis: a case report and review of the literature. PMID- 9850507 TI - Case report: CMV retinitis in two 10-month-old children with AIDS. PMID- 9850509 TI - Viruses and virus-like particles identified in ostrich gut contents. AB - Samples of either gut content from ostriches showing symptoms of enteritis, or allantoic fluid of eggs inoculated with ostrich isolates, were examined for the presence of viral agents by direct negative-contrast electron microscopy. Only a few virus types could be identified with certainty, namely a circovirus (1 sample), a coronavirus (1 sample), a member of either the toga- or bunyaviridae (1 sample), enterovirus (16 samples) and paramyxovirus (26 samples). A large number of samples contained structures resembling myxovirus particles that were interpreted as fringed membranous particles of non-viral origin. An unusual observation of probable single-strand nucleocapsid helices, possibly originating from digested plant material and which were identified in a number of small intestine samples, is reported. This is the 1st report of a spectrum of viruses and virus-like particles occurring in enteric samples from ostriches in South Africa. The low incidence and variety of viruses reported here contribute to the multifactorial origin and complexity of enteric disease in ostriches as well as in other birds and mammalian species. PMID- 9850508 TI - Case report: essential iris atrophy in the Williams-Beuren syndrome. PMID- 9850510 TI - The effect of dietary protein on reproduction in the mare. V. Endocrine changes and conception during the early post partum period. AB - Pregnant Anglo-Arab and Thoroughbred mares (n = 24) were divided randomly according to age and breed into 4 groups of 6 mares each from approximately 6 weeks before their expected foaling date. Diets received by the 4 groups varied in essential amino-acid and total protein contents. Serum progestagen, FSH and LH concentrations were determined from the day of parturition until foal heat and during the 1st oestrous cycle following foal heat. Serum progestagen, FSH and LH concentrations did not differ between the treatment groups. Progestagen concentrations were high (mean = 7.0: 5.2-16.4 ng/ml) at parturition but decreased rapidly within 48 h. As progestagen concentrations decreased LH concentrations increased from Days 3-6 post partum to reach maximum values at, or the day after ovulation. FSH concentrations declined 3-4 d after parturition and increased 2-3 d before ovulation at foal heat. The duration of elevated progestagen concentrations during the luteal phase of the subsequent oestrous cycle affected the interovulatory period. A 12-14 d FSH cyclical releasing pattern occurred. Season/photoperiod affected the resumption of normal oestrous cyclicity during the post partum period. The duration of the 1st oestrous cycle after foal heat in mares fed a low-quality protein diet showed a greater range (13-30 d) compared to mares fed a high-quality protein diet (18-26 d). PMID- 9850511 TI - Observations on the use of midazolam for sedation, and induction of anaesthesia with midazolam in combination with ketamine in the goat. AB - Midazolam hydrochloride administered intramuscularly at a dosage of 0.4 mg/kg induced sedation and sternal recumbency in goats. Increasing the dosage to 1 mg/kg resulted in rapid onset of ataxia followed by lateral recumbency, and loss of consciousness. Light surgical anaesthesia lasted for a period of 7-15 min and was suitable for non-painful procedures. Heart rate was significantly increased (p < 0.05) at both dosage rates, while respiration rate was only increased after midazolam at 0.4 mg/kg. The combination of midazolam (0.4 mg/kg) and ketamine hydrochloride (4 mg/kg) increased heart and respiration rate significantly (p < 0.05). A light plane of surgical anaesthesia suitable for endotracheal intubation was induced, which lasted for a period of 16-39 min. PMID- 9850512 TI - Seasonal occurrence of coccidia in a mixed herd of sheep and goats at Nebo, northern province, South Africa. AB - The seasonal occurrence of coccidial oocysts was determined in a mixed herd of indigenous goats and cross-bred sheep at Nebo, Northern Province, South Africa. The herd of c. 30 sheep was housed with c. 70 goats in 1 camp. Over a period of 14 months, 114 faecal specimens were collected from sheep and 369 from goats and the number of oocysts per gram of faeces (OPG) determined. The mean OPG value for the adult sheep (1187.9) was significantly (p = 0.035) lower than the mean OPG value for the adult goats (7042.9). The highest mean value was recorded in the adult sheep during March 1996 (3044.4) and in adult goats during June 1995 (51568.7). PMID- 9850513 TI - Ketamine hydrochloride--an adjunct for analgesia in dogs with burn wounds. AB - The management of pain in patients with burn wounds is complex and problematic. Burn-wound pain is severe, inconsistent and underestimated. Patients experience severe pain, especially during procedures, until wound healing has occurred. A multi-modality approach is needed for effective management of pain, which requires an understanding of the mechanisms of pain. Altered pharmacokinetics and pharmacodynamics in burn-wound patients makes drug actions unpredictable. Opioids alone are seldom sufficient for pain control. The multi-modality approach includes the use of opioids and non-steroidal anti-inflammatory, anxiolytic and alternative drugs. Ketamine has been found to be a useful agent for analgesia in burn-wound patients; a dose of 10 mg/kg qid per os was found to be an effective adjunct to pain therapy. PMID- 9850514 TI - Tuberculosis in Kafue lechwe (Kobus leche kafuensis) and in a bushbuck (Tragelaphus scriptus) on a game ranch in central province, Zambia. AB - Mycobacteriosis was diagnosed for the first time outside a national park in free ranging wild animals on a game ranch in Zambia. A Kafue lechwe (Kobus leche kafuensis) was found dead with tuberculous lesions on a ranch near Lusaka. Acid fast bacilli were found in the affected organs. Mycobacteria were isolated from these tissues. A bushbuck (Tragelaphus scriptus) was found dead on the same ranch with multiple superficial abscesses in the neck region, extensive granulomatous lesions in the lung, the bronchial and mediastinal lymph nodes and several nodular lesions in the spleen. Few acid-fast bacilli were found in the exudate from the abscesses and lesions in the affected organs. Histologically the lesions resembled those of tuberculosis, but mycobacteria could not be isolated. In addition, 1 Kafue lechwe among 37 wild ungulates of 13 species shot on the ranch showed typical tuberculous lesions in the lungs, but the diagnosis was not confirmed by bacterial isolation. The role of the Kafue lechwe as maintenance host for tuberculosis as well as in the possible spread of this disease to other wildlife are discussed. PMID- 9850515 TI - Atypical dermoid sinus in a chow chow dog. AB - A case of multiple dermoid sinuses in the dorsal cervical and craniothoracic regions in an adult chow chow dog is described. One sinus did not open on the skin surface. This is the first reported case of the condition in this breed and the first time absence of the sinus opening on the skin is described. The use of the term pilonidal sinus to describe this condition is challenged. PMID- 9850516 TI - A case of ovine lymphosarcoma in Kenya. AB - Ovine lymphosarcoma was diagnosed clinically in a 5-year-old Blackhead sheep in Kenya, and confirmed on blood smear, leukocyte count and macroscopic pathology. Four progeny of the ewe were examined. They were normal haematologically but 2 revealed bilaterally enlarged lymph nodes. PMID- 9850517 TI - Parotid duct laceration repair in two horses. AB - Repair of parotid duct lacerations in 2 horses is described using intraluminal silastic tubing as a stent. The duct was lacerated traumatically at the facial vessel notch (incisura vasorum facialium) in the 1st horse, and iatrogenically after removal of an intraluminal sialolith after development of infection within the duct in the 2nd horse. In both cases, a silastic tube was passed retrograde into the duct via the salivary papilla, past the wound until the end lay rostroventral to the parotid salivary gland. The severed salivary ducts and the wounds were sutured. The external portion of the silastic tube was sutured to the skin and the tube left in place. Recovery in the 1st case was uneventful. In the 2nd case a salivary duct/cutaneous fistula formed at a wound distant from the sutured wound, which healed spontaneously. This technique differs from a similar described technique in that the stent tube exits the oral cavity and is attached to the outer skin surface. PMID- 9850518 TI - Clinical aspects of an outbreak of papillomatous digital dermatitis in a dairy cattle herd. AB - Digital skin lesions and lameness of several weeks duration, with a morbidity rate of 28.3%, was reported in a group of 60 Holstein-Israeli dairy cows in various stages of lactation. A clinical survey was performed to monitor recovery and to confirm eradication of bovine papillomatous digital dermatitis in the herd. The combined effects of intensive individual treatment of the 4 lame cattle with procaine penicillin and metronidazole, and subjecting all animals in the herd to a foot bath with a solution composed of formaldehyde and sodium hydroxide twice a week for 12 weeks, were found to achieve a dramatic positive response in all affected cows in the herd. During a 1-year follow-up period no recurrence and/or new cases have been diagnosed. PMID- 9850520 TI - Patent tactics in molecular diversity: things that get around, come around (or: there are always exceptions to the rule). AB - Certain activities which would constitute infringement of a U.S. patent can come within a statutory exemption when the activity is carried out to obtain FDA approval. Exemptions to infringement provided by 35 U.S.C. section 271(e)(1) are discussed along with recent cases interpreting this statute. The evolution of the statute in providing an exemption to activities necessary to obtain FDA marketing approval is described to provide background, legislative intent and show a comparative of the law before and after 1984 when 271(e)(1) was enacted. By analyzing recent cases, criteria for qualifying an activity as exempt from infringement are described--as are activities that do not qualify for the 271(e)(1) exemption. Overall there appears to be a trend to more broadly allowing for activities to qualify as exempt from infringement. This trend may affect the value of patents and determine which technologies a company will seek patent protection on. PMID- 9850519 TI - Comprehensive survey of chemical libraries yielding enzyme inhibitors, receptor agonists and antagonists, and other biologically active agents: 1992 through 1997. AB - This review is a historical accounting of chemical libraries from which biologically active agents have been obtained. The comprehensive tabulation includes citations as early as 1992, when the first descriptions of biologically active libraries were disclosed, and continues through 1997. Four tables are provided listing libraries screened against (1) proteolytic enzymes, (2) non proteolytic enzymes, (3) G-protein coupled receptors (GPCRs), and (4) other targets not classified in the first three tables (e.g. non-GPCRs, integrins, antiinfectives). A name, generic structure, and size is provided for each library citation, accompanied by the molecular screen and the structure and potency of the most active library member. In total, 86 libraries are presented with 60% of the contributions reported from pharmaceutical and biotechnology companies. Approximately 70% of the libraries have used alpha-amino acid synthons in their construction and 85% of the libraries include one or more amide bonds. PMID- 9850521 TI - Patent strategies in molecular diversity. AB - The patentability of combinatorial libraries is examined, requirement by requirement. In general, combinatorial libraries can be patented, as long as the library meets the statutory criteria of utility, novelty, and non-obviousness, and the application meets the standards of enablement, best mode, and written description. Licensing and alternatives to patenting are also considered, along with potential problems unique to combinatorial chemistry agreements. PMID- 9850522 TI - A new method for the characterization of chemical libraries--solely by HPLC retention times. AB - A method has been developed for the assignment of HPLC peaks to their corresponding compounds in libraries of single compounds (parallel syntheses). The basis of the new method is the correlation of the product retention times with the different substituents in the variable positions of the molecule. The correlation is performed automatically by a new algorithm which is part of the computer program LIBFINDER. This practical, easy-to-use tool accelerates the analysis, characterization and purification of chemical libraries, without the need for expensive HPLC-MS equipment. PMID- 9850523 TI - Synthesis of hydantoins via N,N'-ureas derived from polymer-bound amino acids. AB - Starting from carboxy-linked amino acids on trityl functionalized polystyrene resin a highly efficient solid-phase synthesis of hydantoins via N,N'-ureas was elaborated. The polymer-bound hydantoins can be used as a scaffolds for further combinatorial transformations, such as alkylation. Cleavage from the resins yielded the corresponding hydantoins in good yields and purities as shown by ESI MS and HPLC. PMID- 9850525 TI - Tolerance of the fetus by the maternal immune system. Current concepts. PMID- 9850524 TI - Synthesis and characterization of cyclic pseudopeptide libraries containing thiomethylene and thiomethylene-sulfoxide amide bond surrogates. AB - We describe the first examples of a series of cyclic pseudopeptide libraries that have been prepared in a systematic approach in order to facilitate both synthesis and subsequent deconvolution attempts. Our synthetic strategy involved the attachment of a trifunctional amino acid (Asp, Asn or Glu) to a polystyrene resin via its side chain, and stepwise chain elongation using either protected amino acids or a pseudodipeptide building block. Head to tail cyclic peptides were formed by removal of the temporary N- and C-terminal protecting groups followed by ring closure by amide formation. Cyclization of the hexa, hepta, and octapseudopeptides on the resin avoided dimer formation, as monitored by mass spectrometry. We utilized a 'psi-scan' approach in which a second fixed position was serially addressed by stepping a dipeptide surrogate, Pro psi [CH2S]Gly around the rings to generate a group of cyclic pseudopeptide sub-libraries. Oxidation of psi [CH2S] to psi [CH2SO] helped validate the synthesis and also provides a strategy for forming a new set of pseudopeptide libraries (previously described as 'libraries from libraries'). Our results suggest that libraries of cyclic pseudopeptides are an efficient method of preparing and assaying these synthetically more challenging entities as potential drug leads. PMID- 9850526 TI - HLA-G and pregnancy. PMID- 9850527 TI - Fetomaternal HLA differences and the onset and course of rheumatoid arthritis. PMID- 9850528 TI - Animal models of antiphospholipid syndrome. PMID- 9850529 TI - 1998 update on pregnancy in lupus patients. New horizons, new hopes. PMID- 9850530 TI - Antimicrobial therapy in inflammatory joint disease. PMID- 9850531 TI - Value of magnetic resonance imaging in synovitis. PMID- 9850532 TI - Cyclooxygenases and cyclooxygenase inhibitors: is a new class of drugs in the making? PMID- 9850533 TI - Estrogens and osteoporosis. PMID- 9850534 TI - Glucocorticoid therapy in common lumbar spinal disorders. PMID- 9850535 TI - New approaches to the development of virus vaccines for veterinary use. AB - The marked progress in recombinant deoxyribonucleic acid (DNA) technology during the past decade has led to the development of a variety of safe new vaccine vectors which are capable of efficiently expressing foreign immunogens. These have been based on a variety of virus types--poxviruses, herpesviruses and adenoviruses--and have led to the production of many new potential recombinant vaccines. Of these recombinant vaccines, the rabies vaccine, in which the rabies G protein is expressed in a vaccinia vector, has been widely used in the field to prevent the spread of rabies both in Europe and in the United States of America. A recombinant Newcastle disease virus vaccine, using fowlpox virus as the vector to express immunogenic proteins from the Newcastle disease virus, has been licensed as the first commercial recombinant vectored vaccine. Many other recombinant virus vaccines are still at the stage of laboratory or field testing. The most recent breakthrough in vaccinology has been the success with the use of naked DNA as a means of vaccination. This approach has shown great promise in mouse model systems and has now become the most active field in new vaccine development. Molecular redesigning of conventional ribonucleic acid (RNA) viruses to obtain more stable attenuated vaccines was previously possible only for positive-strand RNA viruses, such as poliovirus. However, recent advances in molecular biological techniques have enabled the rescuing of negative-strand viruses from DNA copies of their genomes. This has made it possible to engineer specific changes in the genomes of Rhabdoviridae and Paramyxoviridae, both of which include several viruses of veterinary importance. The authors describe the current progress in the development of vector vaccines, DNA vaccines and vaccines based on engineered positive- and negative-strand RNA virus genomes, with special emphasis on their application to diseases of veterinary importance. PMID- 9850536 TI - Sensitivity and specificity of serological and bacteriological tests for contagious bovine pleuropneumonia. AB - In 1990 an outbreak of contagious bovine pleuropneumonia (CBPP) occurred in Italy. Subsequent surveillance for CBPP was based on random sampling in bovine herds, serological controls on all animals moved from the herd of origin and controls on slaughtered animals. Official tests employed were the complement fixation test (CFT) and bacteriological isolation and typing. A total of 33,856 serum samples collected from herds in CBPP-free regions were used to define CFT specificity, while samples from 595 animals from infected herds were employed to define the sensitivity. Ninety-nine animals from three infected herds were used to estimate the sensitivity of the isolation technique. Results showed the specificity of CFT (threshold +1:10) to be 98% and sensitivity to be 63.79%. The sensitivity of the test did not change significantly, regardless of whether the lesions were caused by acute or chronic infection. The sensitivity of the isolation technique was 54.1%. PMID- 9850537 TI - [The 1996 epizootic of African swine fever in the Ivory Coast]. AB - African swine fever was reported for the first time in the Cote d'Ivoire on 16 April 1996, in Abidjan. The epizootic resulted in the death of 135,000 pigs, equivalent to 29% of the pig population, of which 80% came from the commercial sector. By November, the outbreaks had stabilised and were then eliminated. All epidemiological evidence showed that the active form of disease ceased in September 1996. A variety of factors were responsible for limiting and then halting progression of the disease from the end of August 1996, including a new strategy which gave priority to local information, better control of the situation, depopulation of the infected area in Abidjan, the epidemiological containment of most cases within the villages, and also the assistance and direct involvement of the Food and Agriculture Organization (FAO) of the United Nations. The contribution of the FAO was decisive. It provided a diagnosis of the situation, recommended emergency measures, developed a control strategy, ensured the implementation of the control programme, set up a diagnostic laboratory unit and provided training for staff, slaughtered and destroyed any remaining pigs, set up and implemented a local information programme, developed and implemented a scheme for sentinel animals and for epidemiological surveillance, and designed, performed and analysed an epidemiological survey. From the commencement of the epizootic, two major shortcomings became apparent: inadequate dissemination of information, and a shortage of technical resources allocated for the control of African swine fever. These two shortcomings were compounded by other technical and socio-economic constraints. PMID- 9850538 TI - Comparison of a reverse transcription-polymerase chain reaction assay and virus isolation for the detection of classical swine fever virus. AB - The authors evaluated the ability of a reverse transcription-polymerase chain reaction (RT-PCR) assay to detect classical swine fever virus (CSFV) in comparison with virus isolation and detection by an indirect immunoperoxidase assay (VI-IPA). To determine the specificity of the assay, samples from 60 spleens, 45 tonsils, ten submandibular lymph nodes, eight mesenteric lymph nodes and four kidneys, collected from pigs of various ages which had been slaughtered in abattoirs in Canada (a population free from CSFV), were tested. All the samples tested gave negative results by both VI-IPA and RT-PCR. A total of 20 samples were passaged in porcine kidney (PK) 15 cells and retested by both assays. All were found to be negative, giving a specificity of 100%. To determine the analytical sensitivity of the assay, a similar comparative study was conducted, using CSFV grown in tissue culture and tonsil tissues from a CSFV infected pig. For both infected tissues and tissue culture fluids, RT-PCR was ten times more sensitive than VI-IPA. Amounts as small as 0.6 infectious units per 100 mg of tissue were detected by RT-PCR, compared to 6 infectious units by VI IPA. Similarly, RT-PCR could detect as little as 0.1 infectious unit per ml in tissue culture fluids, compared to one infectious unit per ml by VI-IPA. To determine diagnostic sensitivity, three coded panels (two internal and one external), comprising 45 samples from 14 pigs, were tested. The diagnostic sensitivity of both RT-PCR and VI-IPA was found to be 100% for both internal panels. The results of the external panel, apart from two samples that were missed by both RT-PCR and VI-IPA, were found to be in total agreement. These two samples remained negative after amplification in PK15 cells. All the RT-PCR results were based on a single test whereas, for the VI-IPA results, positive results were obtained for five samples only after an amplification round in PK15 cells. Application of the RT-PCR assay for the diagnosis of CSFV would enable improved detection of the virus in a shorter time period. PMID- 9850539 TI - EpiMAN-SF: a decision-support system for managing swine fever epidemics. AB - During an outbreak of classical swine fever (hog cholera) or African swine fever, decisions in emergency headquarters have to be made under conditions of time pressure, limited resources and uncertainty. The computer program EpiMAN-SF is designed to support decision-making in this situation by providing up-to-date information and a structured approach to choosing from among competing tasks by combining electronic data management with expert system components. EpiMAN-SF provides accurate and timely summaries of the epidemic, using both text and graphics. In addition, the program provides tools for the epidemiological analysis and forecasting of the emergency, and for assessing current and alternative control strategies. The expert system and simulation models integrated in EpiMAN-SF have been evaluated using sensitivity analysis, historic outbreak data and expert opinion. The results demonstrate that EpiMAN-SF is a valid alternative to traditional data management during exotic disease epidemics. PMID- 9850540 TI - Epidemiological surveillance of rabies in Lithuania from 1986 to 1996. AB - Rabies has been endemic in Lithuania for centuries. The objective of this study was to evaluate rabies incidence and seasonal trends in domestic and wild animals in Lithuania from 1986 to 1996. Annual rabies reports and data on cattle numbers were collected. Descriptive epidemiology and time series data analyses were performed to detect seasonal trends. A total of 1,475 specimens were submitted for rabies testing. Of these, 1,248 (84.6%) were confirmed as rabies cases by immunofluorescence antibody testing. Domestic animals accounted for 73.8% (921/1,248) of all rabies cases, with cattle accounting for 61.1% (563/921) of domestic animal cases. Wildlife cases were reported principally in red foxes (56%; 183/327) and raccoon dogs (31.2%; 102/327). Time series analysis indicated a strong association between the occurrence of rabies in cattle and the season of the year (P = 0.0032), with the highest incidence of rabies occurring in autumn. The incidence declined from 3.4 to 4.8 cases per 100,000 cattle in the mid-1980s to < 1.5 cases per 100,000 cattle in 1994, but increased to 4.2 cases per 100,000 cattle in 1996. However, as the cattle population dramatically decreased (by 54%) over the eleven-year period of the study, the reduction in the total number of rabies cases in cattle was not reflected in the annual incidence rate of cattle rabies. PMID- 9850541 TI - A review of leptospirosis in farm animals in Portugal. AB - This paper presents a review of Leptospira infection in farm animals in Portugal which is based mainly on serological results obtained in the National Veterinary Research Laboratory between January 1987 and December 1993. Serum samples were tested by the microscopic agglutination test, at a minimum dilution of 1:100. Positive titres were obtained in 15.3% of the 9,543 bovine samples examined. Sejroe, Pomona, Hebdomadis, Tarassovi and Icterohaemorrhagiae were the principal serogroups which reacted in the tests. A total of 3,195 pigs were tested, of which 20.2% showed positive reactions. The main serogroups which reacted were Australis, Pomona, Cynopteri, Icterohaemorrhagiae and Autumnalis. Field observations on outbreaks of leptospirosis in pigs, along with data obtained from an experimental infection with serovar mozdok in pregnant gilts suggest that this serovar, rather than serovar pomona, may be causing Pomona group infections in pigs. Serum samples from 5,298 sheep were tested and 3.3% gave positive results. The predominant serogroups involved were Canicola, Pomona, Cynopteri, Sejroe and Icterohaemorrhagiae. From the 1,631 goats examined serologically, 5.0% gave positive results, mainly to serogroups Grippotyphosa, Canicola, Pomona, Icterohaemorrhagiae and Pyrogenes. Only 83 serum samples from horses were obtained, of which 43.4% showed positive titres. Serogroups Australis, Autumnalis, Cynopteri and Pyrogenes were those most commonly found. Serological evidence of leptospirosis in farm animals is widespread in Portugal, particularly in cattle and pigs. Leptospirosis in horses needs to be studied further. In an attempt to provide a general view on the occurrence of leptospirosis in these animal species in Portugal, the present results are compared with results obtained in previous studies and are complemented with both previous and recent bacteriological findings. PMID- 9850542 TI - Epizootics of bovine ephemeral fever on dairy farms in Saudi Arabia. AB - In 1990 and 1996, field veterinarians suspected the clinical occurrence of bovine ephemeral fever among dairy and conventional cattle in different regions of Saudi Arabia. The disease has a seasonal occurrence; it begins in early summer (May) and ends in late autumn (November). The mortality rate is low: 0.3% to 0.6%. The morbidity rate ranged from 5% to 61% within the different age groups of one affected herd in the 1996 outbreaks and from 3.4% to 19% among four affected herds in the 1990 outbreaks. A sudden sharp drop in milk production occurred in lactating animals, some of which had become dry by the end of the outbreaks. Trials to isolate the causative virus in cell culture and in baby mice were unsuccessful. Serum neutralisation tests, which used a cell culture-adapted vaccine strain of bovine ephemeral fever virus as an antigen, revealed the presence of specific antibodies with significantly increased titres in the convalescent sera of affected animals. In addition, the testing of paired sera from non-affected heifers and from both dry and milking cows, performed twice, with an interval of 21 days, revealed the presence of neutralising antibodies. In the 1990 outbreaks, comparative serological studies indicated a high percentage (67.5%; 27/40) of seropositive animals in herds in which bovine ephemeral fever had been previously suspected. No antibodies were detected in animals of herds which had no recorded clinical history of bovine ephemeral fever. Following serological confirmation of the prevalence of bovine ephemeral fever in Saudi Arabia, some dairy farms started using a live imported vaccine to control the disease. This study discusses the epizootiological findings in regard to bovine ephemeral fever, as well as its economic impact on four affected dairy farms in 1990. In addition, the authors evaluate the efficacy of immunoprophylaxis in another dairy herd during the same outbreaks. PMID- 9850543 TI - [Serological prevalence and evaluation of the risk factors of bovine enzootic leukosis in the Bogota savannah and the Ubate and Chiquinquira Valleys, Colombia]. AB - The authors present the results of a study to determine the serological prevalence of the enzootic bovine leukosis virus among dairy cows in the Sabana de Bogota region and the Ubate and Chiquinquira Valleys, the principal dairying areas of Colombia. Samples were taken from 919 animals selected at random from 420 farms in 51 municipalities, in accordance with a statistical sampling procedure developed previously, based on the cattle census maintained by the Animal Health and Disease Control Office of the Instituto Colombiano Agropecuario, and the recommendations for prevalence studies of the Pan American Health Organization. The double gel diffusion technique with gp-51 antigen was used. Serological prevalence averaged 45.28% (ranging from 42.07% to 48.49%) with a confidence level of 95%. In addition, a survey was conducted to determine how much those in charge of herds knew about the disease, and to establish the incidence of certain risk factors possibly associated with distribution of the causal agent. Data obtained from 232 completed questionnaires showed that only 54.6% of farms received professional assistance. Of these, 6.6% received constant assistance, 4.4% received visits every fortnight, 51.8% received monthly visits, 14% received visits every other month and 22.95% received occasional visits. PMID- 9850544 TI - Use of polymerase chain reaction to simultaneously detect and type bovine viral diarrhoea viruses isolated from clinical specimens. AB - The techniques of indirect immunofluorescence (IF), immuno-peroxidase (IP) staining and the one-step reverse transcriptase polymerase chain reaction (RT PCR) were compared for detection of 102 isolates of bovine viral diarrhoea virus (BVDV) in infected cell cultures. The BVDV was obtained from bovine clinical specimens, including sera, buffy coats and tissues, submitted from farms located in the States of Iowa and Wisconsin, United States of America. The IF technique detected 88/102 (86.3%) of the viral isolates, whereas IP staining detected an additional 4 isolates (92/102; 90%). The one-step RT-PCR using primers derived from the 5' untranslated region of the BVDV genome detected 102/102 (100%) of the BVDV isolates. A second-round PCR utilising another pair of PCR primers from the 5' untranslated region, allowed rapid genotyping of BVDV. The procedure used showed that the PCR assay based on the 5' untranslated region of the virus genome is the most sensitive indicator for BVDV detection in cell culture, and is also of considerable epidemiological importance since it allowed rapid genotyping of BVDV isolated from clinical specimens. In addition to detection and genotyping of BVDV isolated from clinical specimens, the RT-PCR procedure can be used for routine screening of locally produced and imported biologicals for BVDV contamination. However, the procedure requires further refinement to enable direct application on the clinical specimen. PMID- 9850545 TI - Outbreaks of Dermatophilus congolensis infection in camels (Camelus dromedarius) from the Butana region in eastern Sudan. AB - Natural Dermatophilus congolensis infection of camels has been reported in Kenya in semi-arid areas. Research is being conducted to discover how widespread the condition is in neighbouring countries with similar eco-climatic conditions. Severe skin infections of camels from the Butana region of Eastern Sudan were examined. The infections were first found in two herds of adult camels, of which 50%-75% of the animals were affected. In the other thirteen herds examined, camel calves were more likely to be infected (34%) than adults (8.9%), and lesions were more severe and involved most parts of the body. The lesions began as hair matting and later developed into hard crusts. The case fatality rate ranged from 10% to 30%. D. congolensis was isolated from the scabs. Camel dermatophilosis was found to be among the most serious problems faced by camel herders in the Butana region. PMID- 9850546 TI - An outbreak of a mixed infection of Dermatophilus congolensis and Microsporum gypseum in camels (Camelus dromedarius) in Saudi Arabia. AB - Although both Dermatophilus congolensis and Microsporum gypseum infections have been reported separately in camels, mixed infection involving both agents has not been reported to date. The authors describe a mixed infection of D. congolensis and M. gypseum in camels reared on a dairy farm in Saudi Arabia. A total of 131 out of 559 camels (23.4%) were affected. Forty-eight camels less than one year of age had discrete, circumscribed, crusty, hairless lesions, found in particular on the neck and forelegs. Eighty-three camels of varying ages had extensive hair matting with crusty, hairless lesions, especially on the flanks. Camel calves and young camels demonstrated a relatively greater amount of skin lesions. D. congolensis and M. gypseum were diagnosed by direct microscopy, isolation and histopathology. PMID- 9850547 TI - Serological survey of selected canine viral pathogens and zoonoses in grizzly bears (Ursus arctos horribilis) and black bears (Ursus americanus) from Alaska. AB - Between 1988 and 1991, 644 serum samples were collected from 480 grizzly bears (Ursus arctos horribilis) and 40 black bears (Ursus americanus) from Alaska, United States of America, and were tested for selected canine viral infections and zoonoses. Antibody prevalence in grizzly bears was 0% for parvovirus, 8.3% (40/480) for distemper, 14% (68/480) for infectious hepatitis, 16.5% (79/480) for brucellosis, 19% (93/480) for tularaemia and 47% (225/478) for trichinellosis. In black bears, prevalence ranged from 0% for distemper and parvovirus to 27.5% for trichinellosis and 32% for tularaemia. Antibody prevalence for brucellosis (2.5%) and tularaemia (32%) were identical for grizzly bears and black bears from the geographical area of interior Alaska. Links between differences in prevalence and the origin of the grizzly bears were observed. Antibodies to canine distemper virus and infectious hepatitis virus were mainly detected in grizzly bears from Kodiak Island and the Alaskan Peninsula. Brucellosis antibodies were prevalent in grizzly bears from western and northern Alaska, whereas tularaemia antibodies were detected in grizzly bears from interior Alaska and the Arctic. There was a strong gradient for antibodies to Trichinella spp. from southern to northern Alaska. For most diseases, antibody prevalence increased with age. However, for several infections, no antibodies were detected in grizzly bears aged from 0 to 2 years, in contrast to the presence of those infections in black bears. Grizzly bears served as excellent sentinels for surveillance of zoonotic infections in wildlife in Alaska. PMID- 9850548 TI - [Cost analysis of an official campaign for animal disease control: the example of Senegal in 1996]. AB - A realistic estimation of the cost of government campaigns to control rinderpest and contagious bovine pleuropneumonia is essential in order to determine profitable fees for private veterinarians with a health mandate in Senegal. A cost analysis of a vaccination campaign, organised by the Veterinary Services in Senegal between 1995 and 1996, involving 547,735 cattle, was conducted. Revenue and total costs were 50 CFA francs and 110 +/- 37 CFA francs per head, respectively. Staff-related costs accounted for 39.41% +/- 7.62% of the total cost. The gross and net margins corresponded to the average subsidies of 42 +/- 27 CFA francs and 60 +/- 37 CFA francs, respectively. Simulations showed better relative efficiency for private veterinarians who vaccinate at a price of between 97 and 84 CFA francs per head. Economies of scale varied from 8 to 13 CFA francs per head when the vaccination team increased the number of animals initially vaccinated (16,000 head) by 25% and 44%, respectively. This indicates that a fee of 125 CFA francs per head should guarantee the profitability of compulsory vaccination for private veterinarians. PMID- 9850549 TI - Observations on African horse sickness in Saudi Arabia. AB - The present epidemiological status of African horse sickness in Saudi Arabia, as shown by seroconversion, virus isolation and clinical observation of sentinel horses is described. No African horse sickness virus activity was detected throughout the duration of the study (from November 1992 to March 1995). These findings support previous reports that African horse sickness is not endemic in Saudi Arabia. PMID- 9850550 TI - A serological survey of bovine brucellosis in India. AB - A serological survey of brucellosis in cattle and buffalo was performed in 23 States of India. A total of 30,437 bovine samples, comprising 23,284 cattle and 7,153 buffalo (Bubalus bubalis), were screened. The screening initially used the rose bengal plate test: doubtful and positive samples were then titrated in the serum tube agglutination test. The overall prevalence rate of antibodies was 1.9% in cattle and 1.8% in buffalo. In a detailed study of 47 organised farms in the southern State of Karnataka, 207 of 4,995 (4.1%) serum samples from cattle showed titres for brucellosis. This result was in contrast to the low rate of seropositive results reported in cattle owned by individual farmers in Karnataka (0.7% of 2,424 serum samples). In organised farms with a history of abortion, placenta retention and repeat breeding, the prevalence rate was 17%. PMID- 9850551 TI - Implementation of quality assurance in national foot and mouth disease laboratories, based on the guidelines of the Office International des Epizooties. AB - The final act of the Uruguay Round of the General Agreement on Tariffs and Trade, 1994, contains the Agreement on the Application of Sanitary and Phytosanitary Measures, which aims to reduce the negative effects of health barriers on international trade to a minimum. Therefore, the Office International des Epizooties (OIE) Regional Commission for Europe proposed that an accreditation system based on the EN 45000 standard should be applied to achieve international recognition of certificates and testing laboratories. To this end, the OIE Standards Commission published a series of guidelines for the evaluation of laboratory quality and proficiency testing. In 1995, the Food and Agriculture Organization European Commission for the Control of Foot and Mouth Disease recommended that the OIE proposal should be applied in all foot and mouth disease (FMD) diagnostic laboratories in Europe. This review summarises the EN 45001 standard and the OIE guidelines, and proposes that these guidelines should be taken as a basis for the implementation of a quality assurance programme in FMD laboratories. PMID- 9850552 TI - Guide to epidemiological surveillance for rinderpest. Office International des Epizooties. AB - The practical implementation of epidemiological surveillance programmes for rinderpest prescribed by the standards of the Office International des Epizooties (OIE) is described. The rationale of surveillance is discussed in relation to the stages of disease control, discontinuation of vaccination, clinical surveillance and serological surveillance. These stages lead to provisional and confirmed declarations of freedom from disease, and the confirmed declaration of freedom from infection. Practical methods of stratification of livestock populations, and selection of samples of herds and animals within selected herds are explained. The actions to follow any discovery of disease, or any clinical or serological signs of disease, are discussed. In the serological surveillance stage, balancing the number of herds and the number of animals within selected herds tested can result in considerable reduction in the overall number of serum samples to be tested. The methods used in rinderpest surveillance can readily be adapted to surveillance programmes for other diseases. PMID- 9850554 TI - Recommended standards for epidemiological surveillance systems for rinderpest. Office International des Epizooties. AB - A process involving time-bound steps used to verify the transition from the status of freedom from rinderpest to that of freedom from infection with the rinderpest virus is described. The procedure, informally but widely known as the 'OIE Pathway' (OIE: Office International des Epizooties), originated with the report of the 1989 Expert Consultation on Rinderpest Surveillance Systems. The OIE Foot and Mouth Disease and Other Epizootics Commission, with the assistance of experts on the disease, proposed a revision of the recommended standards in the reference document presented at the 66th General Session of the OIE (66 SG/12/CS3 B, Appendix III). During that General Session, the standards were amended and adopted by the OIE International Committee (Resolution No. IX dated 28 May 1998). PMID- 9850555 TI - Fissure penetration and antibacterial effect in vitro of a glass ionomer cement containing chlorhexidine gluconate. AB - Chlorhexidine has been incorporated in different varnishes to provide a slow release system on the tooth surface in order to reduce mutans streptococci. To provide an alternative vehicle for chlorhexidine with better adhesion properties compared to resin-based varnishes, glass ionomer cement (GI) has been suggested. However, one disadvantage for glass ionomers is a longer setting time compared to the resin-based varnishes. The aim with this study was to compare the fissure penetration and antibacterial characteristics of a glass-ionomer cement (GI) with a GI containing chlorhexidine gluconate (GI-CHX), and GI-CHX with added tataric acid (GI-CHX-TA) to reduce its setting time. Antibacterial properties against mutans streptococci were assessed by agar diffusion. GI, GI-CHX and GI-CHX-TA were applied with a microbrush on the occlusal surfaces of 4, 4 and 6 extracted molars respectively. After setting of cements, sections were ground with 1 mm intervals and photographed. The fissure penetration and adaptation of the cements were scored excellent, acceptable or unacceptable under blind conditions according to a standard. Seventy percent scored excellent with GI-CHX-TA (n = 54) compared with 40% with GI-CHX (n = 48) and 38% with GI (n = 40), (p < 0.05). GI CHX and GI-CHX-TA had significant better antibacterial properties compared to GI or GI with added tataric acid only. PMID- 9850556 TI - Restoration of small proximal dentin lesions with the tunnel technique. A 3-year clinical study performed in Public Dental Service clinics. AB - The aim of this study was to evaluate the 3-year performance of tunnel restorations in Public Dental Service clinics in the county of Blekinge, Sweden. The restorations were made at the indication of a radiographically observed small proximal caries lesion extending into the outer part of dentin in permanent premolars and molars. A total of 302 restorations made by 17 dentists were evaluated 1 yr, 2 yrs and 3 yrs after placement using modified USPHS criteria. The preparations were filled with a cermet glass polyalkenoate cement (Ketac Silver, ESPE Gmbh, Germany). Evaluation was carried out by clinical and radiographical means. The percentage of successful and acceptable restorations was 92.7% after 1 yr, 89.5% after 2 yrs and 84.3% after 3 yrs. Eight % of the restorations failed due to caries, 6% fractured and 1% were replaced due to deep margin discoloration. The results indicate that tunnel restoration is a useful technique in small proximal dentin lesions in permanent premolars and molars. PMID- 9850557 TI - Replacements of restorations in the primary and young permanent dentition. AB - The present study focused on the type of restorative material used and the reasons for replacements of restorations in the primary and the young permanent dentition. All patients with restorations and who were 8 and 19 years of age in 1995 and were regularly treated at 11 Public Dental Health clinics in Jonkoping County, Sweden, participated in the study. Data were extracted from the records for all types of restorations in canines and molars for the preceeding of 5 years for the 8-year-olds (i.e. from 3 to 8 years of age; n = 546) and for approximal restorations in premolars and molars for the preceeding 13 years for the 19-year olds (i.e. from 6 to 19 years of age; n = 606). In all, 6012 restorations were evaluated. The two most common restorative materials used in the primary dentition were compomer and glass ionomer cement and in the young permanent dentition composite and amalgam. In the primary dentition, 29% of the restorations had been replaced and 4% of the teeth with restorations had been extracted. Thus, 33% of the restorations in the primary dentition failed. The corresponding figure for the young permanent dentition was 13%. The most common reason for replacements in the permanent dentition was secondary caries. That restorations often fail because of caries and that the development of secondary caries is not prevented by replacement of an old restoration indicate that more attention should be paid to preventive dental care for patients with restorations in the primary as well as in the young permanent dentition. PMID- 9850558 TI - Number and frequency of bitewing radiographs and assessment of approximal caries in 14-to-19-year-old Swedish adolescents. AB - The hypothesis of the present study was that previous experience of caries may serve as an indicator for individualised bitewing examinations in 15-19-old adolescents. One hundred adolescents, all 19 years old, were randomly selected from the files of public dental clinics in the County of Orebro. For 93 adolescents the total number of radiographs were summed for every patient from the age of 3 to 19 by assessing the files of public dental clinics and by assessing the files of the dental radiology department. All sets of bitewing radiographs from 14 up to and including the age of 19 were assessed with respect to approximal caries. The average number of intraoral radiographs exposed in a patient from the age of 3 through 19 was 24.4. It was shown that 70 to 80% of 14 19-year-old-adolescents had had a bitewing examination every year. The decision to perform a bite-wing examination in a 15-year-old was significantly correlated to the number of surfaces with initial caries at the age of 14. In the other age groups, none of the investigated variables was found to be significantly correlated to the performance of radiographic examinations. PMID- 9850559 TI - Tooth loss and periodontal bone level in individuals of Jonkoping County. A comparison between two adult populations living in the city and in the surrounding area. AB - Cross-sectional and longitudinal studies were performed in the community (in this paper changed to city) of Jonkoping, Sweden, over a period of 20 years to follow changes in oral health and oral health behaviour. To widen our knowledge about dental health and dental care among the adult population, we expanded the study in 1993 to cover the whole county. The specific aim of the present study was to describe tooth loss (excluding third molars) and periodontal bone level in adult residents of Jonkoping County and to compare these two parameters in adults living in the city of Jonkoping with the same in adults living in the rest of the county. Random samples of individuals 30, 40, 50, 60, and 70 years old were selected. A total of 484 persons from the city and 1219 subjects from the rest of the county were examined. A total of 32 (7%) and 27 (2%) persons were completely edentulous in the examined populations from the city and from the rest of the county, respectively. A majority belonged to the older age groups, 60 and 70 years, with 17% of the subjects in the city being edentulous compared with 13% in the rest of the county. The mean number of missing teeth in subjects in the city versus subjects in the rest of the county was 0.75/0.95, 1.37/1.60, 3.34/2.43, 6.34/7.40, and 9.95/10.26 in 30-, 40-, 50-, 60-, and 70-year-olds, respectively. Of all the different tooth types, the average number of molars per person decreased the most with increasing age from an average of 7.79/7.83 (city/county) to 3.06/3.09 (city/county) for 30- and 70-year-olds, respectively. The proportion of subjects without molars was higher in the older age groups in both the city and the rest of the county with 4.8/10.7% and 15.6/22.0% of the 60- and 70-year olds, respectively, lacking molars. In both populations, the mean periodontal bone level decreased with age. It was concluded that no important differences in tooth loss and periodontal health could be seen between the two populations. When organising dental care, dental health administrators could apply the findings from the population in the city to the entire county. PMID- 9850560 TI - Modulation of episodic adrenocorticotropin hormone secretion by cadmium in male rats. AB - The effects of cadmium on adrenocorticotropin hormone (ACTH) secretion are controversial and seem to depend on the dose and duration of the exposure to the metal. This work was undertaken to analyze the effects of acute cadmium administration on the episodic pattern of ACTH release in adult male rats. For this purpose, animals were cannulated 40 h before the experiment to allow a continuous blood withdrawal. Two and a half hours after the administration of a single dose of cadmium chloride (4.5 mg kg-1 bodyweight), the episodic pattern of ACTH was analyzed during three hours (from 10:30 to 13:30, samples being collected every seven minutes) in conscious and freely moving adult male rats. The mean values of ACTH during the bleeding period and the absolute pulse amplitude were decreased by acute cadmium chloride administration (P < or = 0.001, P < or = 0.01, respectively). By contrast, the frequency of ACTH pulses increased (P < or = 0.01). However, no changes in any other parameters of episodic ACTH secretion were observed compared with control animals. These data suggest that cadmium interferes with the regulatory mechanism of ACTH. PMID- 9850561 TI - Lead exposure in children: levels in blood, prevalence of intoxication and related factors. AB - Lead is a highly toxic metal, the main source of which is contamination from combustion of unleaded petrol. The aims of this work were to detect the degree of lead exposure in a large sample of children; determine the relationship between blood lead levels (BPb) and age, sex, habitat and season of the year; and correlate BPb with zinc protoporphyrin (ZPP) values. A cross-sectional study was carried out. Blood from routine extractions drawn at our centre was used. BPb and ZPP were measured by atomic absorption spectrophotometry and haematofluorimetry, respectively. We analysed 1158 blood samples from children. BPb (mean +/- SEM): 0.22 +/- 0.04 mumol l-1. Correlation BPb-age: BPb = 0.19 + 0.086 x age (months), r = 0.129, P < 0.0001. BPb was greater in boys (0.23 +/- 0.007 versus 0.20 +/- 0.006 mumol l-1, P < 0.0002). No differences were observed between habitats (urban versus rural). BPb were higher in the warm months (0.24 +/- 0.013 versus 0.21 +/- 0.007 mumol l-1, P < 0.0001). Prevalence of lead intoxication (BPb > 0.48 mumol l-1) was 4.2%. No differences in prevalence were found among the different groups. The correlation between BPb and ZPP showed r = 0.0969, P = 0.0024. Utility for screening: sensitivity of 53.7% and specificity of 59.3% (cut off point of 60 mumol ZPP mol-1 haem). We can conclude that lead exposure in children in our sample was in the range reported in similar studies in other areas and countries, and below the toxic limit. None of the factors analysed significantly influenced lead intoxication prevalence. There was no good correlation between ZPP and BPb in our samples and the ZPP cut-off point used did not present good specificity and sensitivity values. PMID- 9850562 TI - Stimulatory effect of ascorbate on iron transfer from bleomycin to apotransferrin. AB - The chemotherapeutic agent, bleomycin, forms a 1:1 complex with both Fe(III) and Fe(II). The rate of ferric ion transfer from bleomycin to apotransferrin is rather slow. However, when ascorbate was added to Fe(III)-bleomycin prior to exposure to apotransferrin, the transfer rate was markedly increased. Ascorbate readily reduces Fe(III)-bleomycin to Fe(II)-bleomycin. A second order rate constant of 2.4 mM-1 min-1 was estimated for this reaction. Fe(II)-bleomycin immediately combines with O2, generating the so-called 'activated bleomycin' complex. The data suggest that a reduced form of iron-bleomycin more readily donates its iron ion to apotransferrin. Reoxidation of ferrous ions, and Fe(III) transferrin formation occur rapidly. PMID- 9850563 TI - Decrease of cytochrome c oxidase protein in heart mitochondria of copper deficient rats. AB - Copper deficiency has been reported to be associated with decreased cytochrome c oxidase activity, which in turn may be responsible for the observed mitochondrial impairment and cardiac failure. We isolated mitochondria from hearts of copper deficient rats: cytochrome c oxidase activity was found to be lower than in copper-adequate mitochondria. The residual activity paralleled copper content of mitochondria and also corresponded with the heme amount associated with cytochrome aa3. In fact, lower absorption in the alpha-band region of cytochrome aa3 was found for copper-deficient rat heart mitochondria. Gel electrophoresis of protein extracted from mitochondrial membranes allowed measurements of protein content of the complexes of oxidative phosphorylation, revealing a lower content of complex IV protein in copper-deficient rat heart mitochondria. The alterations caused by copper deficiency appear to be specific for cytochrome c oxidase. Changes were not observed for F0F1ATP synthase activity, for heme contents of cytochrome c and b, and for protein contents of complexes I, III and V. The present study demonstrates that the alteration of cytochrome c oxidase activity observed in copper deficiency is due to a diminished content of assembled protein and that shortness of copper impairs heme insertion into cytochrome c oxidase. PMID- 9850564 TI - Metal ion binding to calmodulin: NMR and fluorescence studies. AB - Calmodulin is an important second messenger protein which is involved in a large variety of cellular pathways. Calmodulin is sensitive to fluctuations in the intracellular Ca2+ levels and is activated by the binding of four Ca2+ ions. In spite of the important role it plays in signal transduction pathways, it shows a surprisingly broad specificity for binding metal ions. Using 15N-Gly biosynthetically-labelled calmodulin, we have studied the binding of different metal ions to calmodulin, including K+, Na+, Ca2+, Mg2+, Zn2+, Cd2+, Pb2+, Hg2+, Sr2+, La3+ and Lu3+, by 1H,15N HMQC NMR experiments. The effects of these ions on the substrate-binding ability of calmodulin have also been studied by fluorescence spectroscopy of the single tryptophan residue in a 22-residue synthetic peptide encompassing the skeletal muscle myosin light chain kinase calmodulin-binding domain. Most of these metal ions can activate a calmodulin target enzyme to some extent, though they bind to calmodulin in a different manner. Mg2+, which is of direct physiological interest, has a distinct site preference for calmodulin, as it shows the highest affinity for site I in the N terminal domain, while the C-terminal sites III and IV are the high affinity binding sites for Ca2+ (as well as for Cd2+). At a high concentration of Mg2+ and a low concentration of Ca2+, calmodulin can bind Mg2+ in its N-terminal lobe while the C-terminal domain is occupied by Ca2+; this species could exist in resting cells in which the Mg2+ level significantly exceeds that of Ca2+. Moreover, our data suggest that the toxicity of Pb(2+)--which, like Sr2+, binds with an equal and high affinity to all four sites--may be related to its capacity to tightly bind and improperly activate calmodulin. PMID- 9850565 TI - Selenium metabolism in Escherichia coli. AB - Escherichia coli will reduce selenite (SeO3(2-)) and selenate (SeO4(2-)) to elemental selenium Se0. Selenium will also become incorporated into proteins as part of the amino acids selenocysteine or selenomethionine. The reaction of selenite with glutathione produces selenodiglutathione (GS-Se-GS). Selenodiglutathione and its subsequent reduction to glutathioselenol (GS-SeH) are likely the key intermediates in the possible metabolic fates of selenium. This review presents the possible pathways involving selenium in E. coli. Identification of intermediates and potential processes from uptake of the toxic oxyanions through to their detoxification will assist us in understanding the complexities of metalloid oxyanion metabolism in these bacteria. PMID- 9850566 TI - Properties of the membranes containing the particulate methane monooxygenase from Methylosinus trichosporium OB3b. AB - The particulate methane monooxygenase (pMMO) from Methylosinus trichosporium OB3b was partially purified and characterized by measuring the effects of reducing agents and additives, and the stability of pMMO was studied. Duroquinol was a suitable reducing agent, and pMMO was stabilized by bovine serum albumin (BSA). Among the additives, the copper (II) ion stimulated pMMO at low concentration and inhibited at high concentration. The optimum conditions for pMMO activity were as follows: 45 degrees C, pH 6.5 and 55 mM 3-morpholinopropanesulfonic acid (MOPS) buffer, and the rate of propene epoxide formation was 13.6 nmol min-1 mg-1 protein. ESR spectra indicate that the copper cluster in the membrane fraction is reduced by duroquinol and oxidized by dioxygen. The result suggests that the copper cluster is contained in the active site of pMMO. PMID- 9850567 TI - Cadmium does not inhibit pulsatile prolactin secretion through TRH. AB - This work was undertaken to analyse the effects of acute cadmium administration on the pulsatile pattern of prolactin release, in adult male rats. For this purpose, animals were cannulated 40 h before the experiment to allow a continuous blood withdrawal. Two hours after the administration of one dose of cadmium chloride (4.5 mg kg1), the pulsatile pattern of prolactin, during three hours, was studied. The effects of two pulses of thyrotropin-releasing hormone (TRH) (1 microgram per rat), given 60 and 120 min after starting the period of blood sampling, were studied. The mean values of prolactin during the bleeding period and the absolute pulse amplitude were decreased by acute cadmium chloride administration. However, no changes in any other parameters of prolactin pulsatility were observed. TRH administration to control rats increased mean prolactin levels, and absolute and relative pulse amplitudes, but decreased the mean half-life of the hormone. In animals pretreated with cadmium, TRH increased the mean levels of prolactin, and absolute and relative amplitudes of the hormone pulses. No other parameter studied was changed by TRH in cadmium pretreated rats. These data suggest that acute administration of cadmium did not inhibit the pulsatile prolactin release through TRH. PMID- 9850569 TI - Metal ion requirement of bifunctional UDP-N-acetylglucosamine 2-epimerase/N acetylmannosamine kinase from rat liver. AB - The metal ion requirement for both enzymatic activities of the bifunctional UDP-N acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (E.C. 5.1.3.14/2.7.1.60), the key enzyme of N-acetylneuraminic acid biosynthesis in rat liver, was investigated. UDP-N-acetylglucosamine 2-epimerase was active in imidazole/HCl buffer in the complete absence of any metal ion. 200 mM Na+, K+, Rb+ and Cs+ activated enzyme activity up to five-fold, whereas lower concentrations of these monovalent metal ions showed only a small effect on UDP-N acetylglucosamine 2-epimerase activity. In sodium phosphate buffer the enzyme activity was increased by 0.5 mM Mg2+, Sr2+, Ba2+ and Mn2+, while in the presence of 200 mM NaCl UDP-N-acetylglucosamine 2-epimerase activity showed a stronger activation by these divalent metal ions. In imidazole/HCl buffer, UDP-N acetylglucosamine 2-epimerase activity was partially inhibited by 0.5 mM Be2+, Mg2+, Ba2+, Mn2+, Sn2+ and Fe2+, and completely inhibited by 0.5 mM Zn2+ and Cd2+. Divalent metal ions were essential for N-acetylmannosamine kinase activity, the most effective being Mg2+, followed by Mn2+ and Co2+. The optimal concentration of these metal ions was 3 mM. Less effective were Ni2+ and Cd2+, whereas Ca2+, Ba2+, Cu2+, Fe2+ and Zn2+ showed no effect on enzyme activity. PMID- 9850568 TI - Novel catecholate-type siderophore analogs based on a myo-inositol scaffold. AB - A novel 1,3,5-triamino-myo-inositol derivative is presented as a readily available scaffold for the design of tripodal siderophore mimetics. Based on this scaffold, various hexadentate catecholate-type siderophore analogs were synthesized by attaching the catechols to the inositol scaffold via spacer units of different structure and length. The potential to tune the polarity of the inositol containing siderophore analogs has also been demonstrated by varying the protection group strategy. The siderophore activity of the prepared siderophore analogs was examined by cross-feeding tests with various Gram-negative bacteria and mycobacteria. PMID- 9850570 TI - Differential cytotoxicity of trace metals in cisplatin-sensitive and -resistant human ovarian cancer cells. AB - Cellular resistance of cisplatin is related to various factors such as membrane transformations, changes in cellular transport systems, and an increased efflux of cisplatin by the tumor cells. Deficiencies of one or more trace metals can affect normal physiological functions, leading to altered enzymatic activities and a reduction in immune responses. This in vitro investigation was undertaken to study and determine the differential cytotoxicity of certain trace metals in human ovarian cancer cells that were sensitive and resistant to cisplatin. Standard cytotoxicity assays were performed using the neutral red assay. In general, the cisplatin-resistant cells exhibited an increased resistance to the externally supplied trace metals. For both cell lines the rank order of cytotoxicity from greatest to least with the non-essential metals was Cd2+ > Bi3+, and for the macrometals, Ca2+ > K+ > Mg2+. The transition metals and selenium exhibited a slight difference between the two cell lines with respect to the order of cytotoxicity. The cisplatin-sensitive cells had a rank order of V5+ > Se6+ > Cu2+ > Zn2+ > Fe3+, from greatest to least toxicity. The cisplatin resistant cells had a rank order of Cu2+ > V5+ > Se6+ > Zn2+ > Fe3+. Since trace metals have various functions in maintaining normal health, these results provide key baseline cytotoxicity data and show that, in general, cytotoxic resistance to the trace metals tested followed a pattern similar to cellular cisplatin resistance. PMID- 9850571 TI - Transferrin response in normal and iron-deficient mice heterozygotic for hypotransferrinemia; effects on iron and manganese accumulation. AB - Hypotransferrinemia is a genetic defect in mice resulting < 1% of normal plasma transferrin (Tf) concentrations; heterozygotes for this mutation (+/hpx) have low circulating Tf concentrations. These mice provide a unique opportunity to examine the developmental pattern and response of Tf to iron-deficient diets, and furthermore, to address the controversial role of Tf in Mn transport. Twenty three weanling +/hpx mice and forty-five wild-type BALB/cJ mice were either killed at weaning or fed diets containing either 13 or 72 mg kg-1 Fe, and killed after four or eight weeks. Plasma Tf concentrations were lower in +/hpx mice, plasma Tf nearly doubled and liver Tf was only 50% of normal in response to iron deficiency. Brain iron concentration did not correlate significantly with either plasma Tf or TIBC. However, iron accumulation into brain continued with iron deficiency whereas most other organs had less iron. These results imply that either there is a selected targeting of iron to the brain by plasma Tf or there is an alternative iron delivery system to the brain. Furthermore, we observed no differences in tissue distribution of 54Mn despite the differences in circulating Tf concentrations and body iron stores; this suggests that there are non-Tf dependent mechanisms for Mn transport. PMID- 9850572 TI - Multiresidue liquid chromatographic method for determining residues of mono- and dibasic penicillins in bovine muscle tissues. AB - A liquid chromatographic method with UV detection at 325 nm was developed for simultaneous determination of amoxicillin, ampicillin, penicillin G, and cloxacillin residues in bovine muscle tissue as their mercaptide derivatives. The penicillins are extracted from bovine tissues with 0.1 M phosphate buffer (pH 8.5), cleaned up on a t-C18 Sep-Pak cartridge, and eluted with 2 mL acetonitrile. After the acetonitrile in the eluate is evaporated to dryness, the residue is dissolved in 200 microL (40 + 60, v/v) acetonitrile-phosphate buffer (pH 6.5) and derivatized with acetic anhydride and mercuric chloride in the presence of 1,2,4 triazole at 65 degrees C for 30 min. Gradient analysis on a Spherisorb 5 microns ODS(2) (octadecyl silane) analytical column using a binary mobile phase consisting of acetonitrile and 0.10 M phosphate buffer (pH 6.5) in the presence of 0.0157 M sodium thiosulfate at 1 mL/min permits determination of each intact penicillin in bovine muscle tissue at > or = 10 ppb with recoveries > or = 72%. This laboratory method provides detection sensitivities equivalent to those of rapid tests used for screening beta-lactam drug residues in bovine tissue samples for regulatory enforcement. PMID- 9850574 TI - Identification of beta-lactam antibiotics in tissue samples containing unknown microbial inhibitors. AB - Antibiotic residues in animal tissues can be detected by various screening tests based on microbial inhibition. In the 7-plate assay used by the U.S. Department of Agriculture's Food Safety and Inspection Service (FSIS), penicillinase is incorporated into all but one plate to distinguish beta-lactam antibiotics from other types. However, beta-lactams such as cloxacillin and the cephalosporins are resistant to degradation by penicillinase. They may not be identified as beta lactams by this procedure, and thus, they may be identified as unidentified microbial inhibitors (UMIs). However, these penicillinase-resistant compounds can be degraded by other beta-lactamases. The present study describes an improved screening protocol to identify beta-lactam antibiotics classified as UMIs. A multiresidue liquid chromatographic procedure based on a method for determining beta-lactams in milk was also used to identify and quantitate residues. The 2 methods were tested with 24 tissue FSIS samples classified as containing UMIs. Of these, 3 contained penicillin G, including one at a violative level, and 5 contained a metabolite of ceftiofur. The others were negative for beta-lactam antibiotics. PMID- 9850573 TI - Solid-phase extraction cleanup and liquid chromatography with ultraviolet detection of ephedrine alkaloids in herbal products. AB - A solid-phase extraction (SPE) cleanup and a liquid chromatographic (LC) method with UV detection is presented for analysis of up to 7 ephedrine alkaloids in herbal products. Alkaloids from herbal products are extracted with acidified buffer, isolated on a propylsulfonic acid SPE column, eluted with a high-ionic strength buffer, and separated by LC with detection at 255 nm. LC separation is performed by isocratic elution on a YMC phenyl column with 0.1 M sodium acetate acetic acid (pH = 4.8) containing triethyl-amine and 2% acetonitrile. Ephedrine alkaloids are completely separated in 15 min. Average recovery of 5 common alkaloids from 3 spiked matrixes is 90%, with an average relative standard deviation (RSD) of 4.4% for alkaloid spikes between 0.5 and 16 mg/g. Average quantitation of ephedrine and pseudoephedrine from 6 herbal products is 97% of declared label claims, and average quantitation of synephrine from an herbal dietary product is 85% of label claim (RSD, 3.2%). Recoveries of synephrine, norephedrine, ephedrine, pseudoephedrine, N-methylephedrine, and N methylpseudoephedrine spiked in 4 herbal products averaged 95%. Results of ruggedness testing and of a second laboratory validation of the procedure are also presented. PMID- 9850575 TI - Determination of apramycin in swine kidney tissue by liquid chromatography with fluorescence detection. AB - A liquid chromatographic method for determining apramycin in swine kidney tissue is described. Apramycin is extracted from tissue with basic methanol and purified by ion-pair extraction. By using an automated derivatization and injection procedure, the purified extract is derivatized with o-phthaldehyde, separated on a C18 column, and detected with a fluorescence detector. For fortified kidney samples, between-run coefficients of variation ranged from 4.8 to 7.1% at 1.00 ppm and from 9.6 to 14.3% at 0.50 ppm. Recoveries ranged from 76 to 86%. Standard curves were linear over the range 10-100 ng/mL. PMID- 9850576 TI - Performance tested method certification of BAX for Screening/Salmonella: a case study. AB - This report details the independent laboratory study of the BAX for Screening/Salmonella assay to complete AOAC Performance Tested Method certification. The performance of the BAX system was compared with those of BAM culture methods on food samples inoculated with Salmonella. This study validated product claims. Performance Tested Method status was granted for the screening assay. PMID- 9850577 TI - Identification of the source of reagent variability in the xanthydrol/urea method. AB - Contamination of food and food packaging material by rodent urine is evidence of insanitary conditions. Urea from rodent urine is used as a chemical indicator of contamination. The limit of detection of the xanthydrol/urea AOAC Method 959.14 by formation of dixanthylurea crystals is 4 micrograms urea isolated from urine on packaging material. Six different lots of xanthydrol from 5 different manufacturers were compared. Differences in urea detection sensitivity of the xanthydrol of up to 1000-fold were observed. Melting points showed further evidence of variability and impurities in xanthydrol lots. A liquid chromatographic method was developed to separate and identify the impurities. Confirmation of analytes was performed by gas chromatography/mass spectrometry. PMID- 9850578 TI - Variability associated with testing shelled corn for fumonisin. AB - Variances associated with sampling, sample preparation, and analytical steps of a test procedure that measures fumonisin in shelled corn were estimated. The variance associated with each step of the test procedure increases with fumonisin concentration. Functional relationships between variance and fumonisin concentration were estimated by regression analysis. For each variance component, functional relationships were independent of fumonisin type (total, B1, B2, and B3 fumonisins). At 2 ppm, coefficients of variation associated with sampling (1.1 kg sample), sample preparation (Romer mill and 25 g subsample), and analysis are 16.6, 9.1, and 9.7%, respectively. The coefficient of variation associated with the total fumonisin test procedure was 45% and is about the same order of magnitude as that for measuring aflatoxin in shelled corn with a similar test procedure. PMID- 9850580 TI - Rapid gas chromatographic method for simultaneous determination of cholesterol and alpha-tocopherol in eggs. AB - A new method was developed for simultaneous determination of cholesterol and alpha-tocopherol in eggs. It involves rapid and simple sample preparation accomplished in one tube and chromatographic separation that does not require derivatization of analytes. Total analysis time per sample is 40 min. Labor, cost, and use of hazardous chemicals are minimized. To ensure selectivity, accuracy, and precision, critical analytical parameters were investigated. Overall recoveries were 98.8 and 99.2% for cholesterol and alpha-tocopherol, respectively. Linearity was acceptable for both analytes (r = 0.9964 for cholesterol and 0.9996 for alpha-tocopherol) in the fortification range examined. Precision data based on within-day and between-days variation gave overall relative standard deviations of 2.0% for cholesterol and 7.0% for alpha tocopherol. The method was applied successfully for quantitation of cholesterol and alpha-tocopherol in eggs. PMID- 9850579 TI - Immunoaffinity column as cleanup tool for a direct competitive enzyme-linked immunosorbent assay of cyclopiazonic acid in corn, peanuts, and mixed feed. AB - An immunoaffinity column (IAC) for cyclopiazonic acid (CPA) was prepared by coupling a CPA-specific monoclonal antibody to CNBr-activated sepharose 4B. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) was used to study the chromatographic behavior of a 0.2 mL gel column with a binding capacity of 4 micrograms CPA/column as well as to evaluate its efficacy as a cleanup tool for analysis of naturally occurring CPA. Sample extract either in buffer solution or in a solution containing up to 35% methanol could be loaded onto the column. After the column is washed with 5 mL deionized water and 5 mL 50% methanol, CPA could be quantitatively eluted with 2 mL 100% methanol. The column could be regenerated at least 10 times by washing with 10 mL equilibrating buffer and then storing in a cold room overnight before reuse. Recoveries of CPA added to corn, peanut, and mixed feed extracts in the range 10-200 ng/g were 88-105, 86-100, and 90-110%, respectively. Detection limits were 2.0, 4.4, and 4.7 ng/g for corn, mixed feed, and peanuts, respectively. Twenty-two peanut samples naturally contaminated with CPA were subjected to both IAC and solvent partition cleanup followed by dc-ELISA. Although a good correlation between data obtained from IAC dc-ELISA and from SP-dc-ELISA (r = 0.75, p < 0.0001) was obtained, the slope of the linear regression was low (0.67), indicating loss during solvent partition cleanup. The overall data showed that the combination of IAC and dc-ELISA is an effective method for CPA analysis. PMID- 9850581 TI - Gas chromatographic determination of azoxystrobin, fluazinam, kresoxim-methyl, mepanipyrim, and tetraconazole in grapes, must, and wine. AB - Azoxystrobin, fluazinam, kresoxim-methyl, mepanipyrim, and tetraconazole were determined in grapes, must, and wine by a gas chromatographic method with nitrogen-phosphorus (NP) and mass spectrometric (MS) detectors. Pesticides were isolated from the matrixes by online microextraction with acetone-hexane (50 + 50, v/v). Because of the high selectivity of NP and MS detectors, no interferent peaks were present and no cleanup was necessary. Recoveries from fortified grapes, must, and wine ranged from 80 to 111%, with coefficients of variation ranging from 1 to 14%. Limits of determination were 0.05 mg/kg for kresoxim methyl and 0.10 mg/kg for the other compounds. PMID- 9850582 TI - Determination of phosphine residues in whole grains and soybeans by ion chromatography via conversion to phosphate. AB - An ion chromatographic (IC) method was developed for determining phosphine (PH3) in whole grains (barley, corn, oats, rice, rye, and wheat) and soybeans. The method converts phosphine to phosphate (i.e., orthophosphate) and isolates the phosphate by IC with eluent-suppressed conductivity detection. Recoveries of unbound phosphine by the method were similar to those obtained by an established colorimetric method for 7 different products fortified at 3 levels. Mean recoveries were low (i.e., 30-60%) and varied with product type and level of fortification. Recoveries of PH3 from previously fumigated products fortified with aluminum phosphide ranged from 19.0% for barley fortified at 0.734 ppm to 88.3% for corn fortified at 1.691 ppm. Precision data from 3 products based on replicate analyses (n = 4 or 5) gave relative standard deviations of 1.78-4.66% for mean laboratory-fumigated PH3 levels of 0.679-1.309 ppm. Estimated limits of detection (LOD) and quantitation (LOQ) for PH3 were 0.010 microgram/g (10 ppb) and 0.0275 microgram/g (27.5 ppb) at signal-to-noise ratios (S/N) of 4:1 and 10:1, respectively. These values were also determined for a nonchemically suppressed IC system with LOD of 0.02 microgram/g (20 ppb) and LOQ of 0.055 microgram/g (55 ppb) at S/N of 4:1 and 10:1, respectively. Phosphate response was linear over the concentration range equivalent to 0.30-10.0 micrograms P/mL, with a mean correlation coefficient of 0.9988 based on replicate standard curves. The relationship of product composition to recovery from various products was also examined. PMID- 9850584 TI - Automated method for cleanup and determination of benomyl and thiabendazole in table-ready foods. AB - An automated solid-phase extraction (SPE) cleanup with on-line liquid chromatographic (LC) analysis was developed to determine residues of benomyl (as carbendazim) and thiabendazole in table-ready food items from the U.S. Food and Drug Administration Total Diet Study (TDS). A strong-cation-exchange cleanup of an acetone extract replaces the methylene chloride solvent partitioning steps in the procedure described in the Pesticide Analytical Manual (PAM). LC analysis is accomplished with a C8 analytical column and tandem fluorescence and UV detection. Recoveries of both analytes from 32 representative TDS foods fortified at 0.05 and 0.5 microgram/g were determined. Method precision was evaluated with triplicate recovery assays on 11 foods fortified at both levels. Accuracy was tested further by assaying 47 foods for incurred residues in parallel with the validated PAM procedure for comparison, and good agreement was found. The automated SPE method reduces solvent consumption, analysis time, and labor. PMID- 9850583 TI - Determination of 17 beta-estradiol and its metabolites in sewage effluent by solid-phase extraction and gas chromatography/mass spectrometry. AB - The paper describes a simple and quantitative method for monitoring non conjugated 17 beta-estradiol (E2) and its metabolites estrone (E1) and estriol (E3) as environmental contaminants in municipal sewage effluents. Estrogens were preconcentrated and cleaned up by solid-phase extraction using a reversed-phase C18 cartridge. They were derivatized with pentafluoropropionic acid anhydride, and the products were analyzed by gas chromatography/mass spectrometry. Recoveries from spiked distilled water and sewage were better than 87% at fortification levels of 100 and 20 ng/L. For a 1 L sewage sample and a concentration factor of 5000, detection limits were 5 ng/L for E1 and E2 and 10 ng/L for E3. In a brief survey of Canadian wastewater, these estrogens were detected in many raw sewage and effluent samples at concentrations ranging from 6 to 109 ng/L for E1, from < 5 to 15 ng/L for E2, and from < 10 to 250 ng/L for E3. PMID- 9850585 TI - Determination of calcium by inductively coupled plasma-atomic emission spectrometry, and lead by graphite furnace atomic absorption spectrometry, in calcium supplements after microwave dissolution or dry-ash digestion: method trial. AB - A 3-laboratory method trial was conducted to evaluate 2 sample digestion procedures and instrumental determination parameters for analysis of calcium and lead in Ca supplements. Calcium supplements were treated by dry-ash digestion or microwave dissolution prior to spectrometric analysis. In each case, Pb was determined by graphite furnace atomic absorption spectrometry and Ca by inductively coupled plasma-atomic emission spectrometry. Blind duplicates of 6 Ca supplement samples were analyzed after each sample treatment procedure. Matrix pairs contained dissimilar Pb levels to cover the analyte range encountered during method development. Calcium content of the Ca supplement samples also reflected the range seen during method development. Stock solutions of Ca and Pb were supplied to collaborators for preparation of quantitation standards to remove a variable external to the method. National Institute of Standards and Technology Standard Reference Material (NIST SRM) 1486, bone meal, was included to assess method accuracy and recovery at NIST certificate Ca and Pb levels for this material (26.58 +/- 0.24% Ca and 1.335 +/- 0.014 micrograms Pb/g). Analyses of the NIST SRM yielded 25.9 +/- 1.1 and 27.2 +/- 2.3% Ca and 1.53 +/- 0.19 and 1.26 +/- 0.19 micrograms Pb/g for dry-ash and microwave procedures, respectively. Statistical analyses of data indicated acceptable repeatability and reproducibility for determination of Pb and Ca in various Ca supplements. With either sample preparation technique, the method is appropriate for determining Pb or Ca in Ca supplements. PMID- 9850586 TI - Monitoring chlorinated pesticides and toxic elements in tissues of food-producing animals in Yugoslavia. AB - According to the established monitoring program in Yugoslavia, 941 swine, 561 cattle, and 358 lamb samples collected during a 5-year period were analyzed for chlorinated pesticide residues. Less than 10% of the examined samples contained residues of HCB (hexachlorobenzene), HCH (hexachlorocyclohexane), lindane (gamma hexachlorocyclohexane), and total DDT (p,p'-DDT and metabolites) at concentrations greater than the lowest detectable limit. None of the swine and cattle samples exceeded the residue limit (RL). Among lamb samples, 2.5% contained lindane residues exceeding the RL, with a mean of 4.75 mg/kg. This finding suggests lindane's improper use as a veterinary pesticide, probably as a sheep-dip for destroying ectoparasites. During the same period, 849 swine, 584 cattle, and 350 lamb samples also were analyzed for toxic elements (Pb, Cd, Hg, and As). In the majority of samples, toxic elements were present at levels less than 50% of RLs. Only Cd and Pb RLs were exceeded in several cases. Differences in trace element contents of samples from different animals were not significant. The data indicate that residues in tissues of food-producing animals do not have a great potential impact on public health. PMID- 9850587 TI - Determination of bitertanol residues in strawberries by liquid chromatography with fluorescence detection and confirmation by gas chromatography/mass spectrometry. AB - A simple and rapid method was developed for determining bitertanol residues in strawberries. Bitertanol was extracted from samples with ethyl acetate. Bitertanol acetate was added prior to extraction as a surrogate standard. The ethyl acetate extract was cleaned up by passing through tandem solid-phase extraction columns consisting of anion-exchange (SAX) and aminopropyl (NH2) bonded silica. The eluate was evaporated to dryness and reconstituted with methanol. Bitertanol residues were determined by liquid chromatography with fluorescence detection. Recoveries at 4 fortified levels (0.05, 0.25, 0.5, and 1.0 microgram/g), calculated by the internal standard method, ranged from 92.1 to 99.1%, with coefficients of variation ranging from 0.3 to 4.0%. The detection limit was 0.01 microgram/g. Of 25 commercial strawberry samples analyzed for bitertanol residues, 5 contained bitertanol residues at concentrations ranging from 0.02 to 0.51 microgram/g. Positive samples were confirmed by gas chromatography/mass spectrometry with mass-selective detection (m/z 170 and 168). PMID- 9850588 TI - A simple method for evaluating data from an interlaboratory study. AB - Large-scale laboratory- and method-performance studies involving more than about 30 laboratories may be evaluated by calculating the HORRAT ratio for each test sample (HORRAT = [experimentally found among-laboratories relative standard deviation] divided by [relative standard deviation calculated from the Horwitz formula]). The chemical analytical method is deemed acceptable per se if HORRAT approximately 1.0 (+/- 0.5). If HORRAT is > or approximately 2.0, the most extreme values are removed successively until an "acceptable" ratio is obtained. The laboratories responsible for the extreme values that are removed should examine their technique and procedures. If > or approximately 15% of the values have to be removed, the instructions and the methods should be examined. This suggested computation procedure is simple and does not require statistical outlier tables. Proposed action limits may be adjusted according to experience. Data supporting U.S. Environmental Protection Agency method 245.1 for mercury in waters (manual cold-vapor atomic absorption spectrometry), supplemented by subsequent laboratory-performance data, were reexamined in this manner. Method performance parameters (means and among-laboratories relative standard deviations) were comparable with results from the original statistical analysis that used a robust biweight procedure for outlier removal. The precision of the current controlled performance is better by a factor of 4 than that of estimates resulting from the original method-performance study, at the expense of rejecting more experimental values as outliers. PMID- 9850589 TI - Improvement in the multiresidue liquid chromatographic analysis of residues of mono- and dibasic penicillins in bovine muscle tissues. AB - A recently developed multiresidue method using gradient liquid chromatographic conditions for analysis of residues of beta-lactam drugs in bovine muscle tissues after precolumn derivatization with acetic anhydride was modified to permit isocratic analysis. The modification included replacing the acylating reagent, acetic anhydride, with benzoic anhydride and increasing sample size for extraction from 2 to 3 g. The modifications have reduced analysis time per sample from > 1 h to about 22 min without loss in detection sensitivities for the beta lactams, resulting in a significant increase in the number of samples that can be analyzed in 1 day from 1 to 8. The modifications make the multiresidue LC method more suitable than the original method for use in regulatory programs for routine analysis of these veterinary drug residues in bovine muscle tissues. PMID- 9850590 TI - Comparison of microwave oven and convection oven for acid hydrolysis of dietary fiber polysaccharides. AB - Hydrolysis of dietary fiber polysaccharides (DFP) is an integral part of any enzymatic-chemical method for dietary fiber analysis. Residues obtained after enzyme treatments of fiber-containing foods are usually suspended in 12 M sulfuric acid and kept at or slightly above ambient temperature for at least 1 h, and then the mixtures are diluted with deionized water to a final concentration of 1 M or 2 M acid, followed by heating at 100 degrees C in a water bath or convection oven for 1 or 2 h. Under these hydrolytic conditions, some degradation of the released monosaccharides generally takes place over the duration of hydrolysis. We investigated the feasibility of using microwave energy as a heat source to reduce time and minimize degradation. Preliminary tests were done on the well-characterized soy polysaccharide Fibrim. With a microwave digestion system equipped with temperature and pressure monitors and control lines, optimum settings of power (5%, 75%), time (up to 3 min and 30 s), temperature (35 degrees 55 degrees C), and pressure (45-65 psi) were determined for different foods depending on the residue weight and volume of acid. Results were comparable for microwave oven and convection oven hydrolysis of DFP from 5 foods with good correlations for neutral sugar values; r2 = 0.997 for arabinose, 0.925 for galactose, 0.981 for glucose, 0.969 for mannose, and 0.990 for xylose. PMID- 9850591 TI - Thin-layer chromatography-mass spectrometry using activated carbon, surface assisted laser desorption/ionization. AB - The analysis of compounds separated by thin-layer chromatography (TLC) by surface assisted laser desorption ionization (SALDI) mass spectrometry has been demonstrated. The compounds are analyzed from the surface of the intact TLC plate, and the preparation of the TLC plate is rapid and robust: the gel surface is covered with 2 microns activated carbon particles, and glycerol is added. Analytes diffuse from the interior of the gel to the surface where they are adsorbed onto the activated carbon. A nitrogen laser is used to desorb analyte ions from the carbon particles in a time-of-flight mass spectrometer. A wide range of organic compounds, including peptides, can be detected, either as protonated or as cationized molecules. Interference with "matrix peaks" is limited since background TLC-SALDI mass spectra typically show only a few intense peaks at low mass. The detection limit for bradykinin from a developed plate is approximately 25 ng (calculated for S/N = 3). The mass resolution (FWHM) varied from a high of about 500 to a low of about 100. This variability was likely due to surface charging. Methods to improve both mass resolution and sensitivity of TLC-SALDI are suggested. PMID- 9850593 TI - Deep dermatophytosis caused by Trichophyton rubrum--a case report. AB - A case of deep dermatophytosis in the gluteal region in a male patient successfully treated with terbinafine is described with its clinical, mycological and histopathological features. PMID- 9850592 TI - Anticryptococcal activity of voriconazole against Cryptococcus neoformans var. gatti vs var. neoformans: comparison with fluconazole and effect of human serum. AB - Voriconazole (VCZ), a new wide-spectrum antifungal triazole currently in development, was tested for activity against Cryptococcus neoformans (CN) var. gattii and var. neoformans in RPMI-1640 (RPMI) or RPMI plus human serum. In RPMI VCZ was 10-fold more inhibitory than FCZ for both varieties of CN. In the presence of human serum neither VCZ nor FCZ had enhanced activity against CN var. gattii. By contrast, both VCZ and FCZ had significantly increased activity in the presence of serum against CN var. neoformans. The lack of serum-enhancing activity for VCZ or FCZ against CN var. gattii may reflect the in vivo situation and predict less efficacy in CN var. gattii infections. PMID- 9850594 TI - Dermatophytes isolated from selected ruminants in India. AB - A survey was carried out in different areas of Uttar Pradesh for isolating and identifying dermatophytes from ruminants. Out of 102 skin scrapings, collected from various animals, 4/22 cattle, 1/9 buffalo, 14/44 sheep and 2/28 goats were KOH positive. In respect of the cattle, the culture positive samples yielded one strain each of Trichophyton mentagrophytes, T. rubrum, T. simii and two strains of Microsporum gypseum. From the buffaloes examined, only one case yielded a dermatophyte and it was identified as T. mentagrophytes. In respect of the sheep, two samples were culture positive and the dermatophyte was T. mentagraphytes (2). Of the 28 goat scrapings, only two harboured dermatophytes viz., M. gypseum and T. mentagrophytes. Fungi other than dermatophytes isolated from the ruminants were Alternaria species, Aspergillus species, Beauveria bassiana, Chrysosporium keratinophilum, Curvularia species, Penicillium species, Pseudoarachniotus roseus and T. terrestre. Isolation of T. rubrum from the skin lesion of one of the cattle is noteworthy, since this dermatophyte is better known as an anthrophilic species. PMID- 9850595 TI - Enumeration and identification of Aspergillus group and Penicillium species in poultry feeds from Argentina. AB - A total of 180 samples of poultry feeds were collected during 1996 and 1997 from different factories in the south of the province of Cordoba-Argentina. They were examined for the occurrence of Penicillium spp. and Aspergillus group species. Likewise, the capacity to produce aflatoxins by the Aspergillus section flavi group was determined. The predominant species of Aspergillus were A. flavus and A. parasiticus. For Penicillium spp., P. brevicompactum, P. purpurogenum and P. oxalicum were identified. Less frequently isolated were A. candidus, A. fumigatus, A. niger, A. orizae, A. parvulus, A. tamarii, A. terreus, and P. expansum, P. funiculosum, P. minioluteum, P. pinophylum, P. restrictum, P. variable and others. The mean value counts ranged from 1 x 10(3) to 9.5 x 10(4) CFU/g for the Aspergillus spp. and from 1.2 x 10(3) to 2.5 x 10(5) CFU/g for the Penicillium spp. When cultured on autoclaved rice kernels for 1 week in the dark at 25 degrees C, mycotoxin production by strains of A. flavus was as follows: 21 of the 45 assayed strains (47%) produced aflatoxins. From them, 24% of the isolates produced AFB1 and AFB2 with levels from 181 to 14545 and 6 to 3640 micrograms/kg respectively. Only 10 strains produced AFB1 with levels from 10 to 920 micrograms/kg. Fifty percent of the A. parasiticus strain was toxicogenic; six aflatoxicogenic profiles were identified. Only 10% of the strains produced all of the aflatoxins. These results showed that a potential exists for the production of mycotoxins by the Aspergillus section flavi and the Penicillium spp. They also suggested an association of mycotoxicosis with poultry feeds in Argentina. PMID- 9850596 TI - Effects of thrombin on the growth, protein synthesis, attachment, clustering and alkaline phosphatase activity of cultured human periodontal ligament fibroblasts. AB - Previous studies have indicated that thrombin can activate pulp cells, including fibroblasts. Because pulp cells and periodontal ligament (PDL) fibroblasts can express thrombin receptor mRNA, the specific aim of this study was to determine whether thrombin can activate the growth, attachment, protein synthesis, alkaline phosphatase activities and cellular clustering of cultured human PDL fibroblasts. Thrombin can stimulate the growth of PDL fibroblasts in a dose dependent manner (as analyzed by MTT assay). At concentrations of 5 and 10 U/ml, thrombin increased the cell numbers to 141% and 153% greater than that of the control after 5 days of incubation, respectively. Thrombin (5-20 U/ml) also stimulated the protein synthesis rate (assayed by [3H]proline incorporation) to 1.88-2.13 fold that of the control. However, pretreatment of PDL fibroblasts with thrombin (1-20 U/ml) could not promote the attachment of PDL fibroblasts to type I collagen and fibronectin. Moreover, thrombin could induce clustering of PDL fibroblasts within a concentration range of 5-20 U/ml. However, thrombin (1-20 U/ml) exerted neither stimulatory nor inhibitory effect on cellular alkaline phosphatase activities. In conclusion, it appears that the presence of thrombin seems to have effects on PDL fibroblasts in terms of cell growth, protein synthesis and cell clustering. This suggests that thrombin might be important in the early healing process of periodontium following periodontal surgery. PMID- 9850597 TI - Transcatheter closure of atrial septal defect guided by on-line transesophageal echocardiography. AB - Transcatheter occlusion of secundum atrial septal defect has been tried since 1976. Some investigators have incorporated on-line transesophageal echocardiography so as to better monitor the procedure. Most, however, have used endotracheal intubation and general anesthesia. The aim of this study was to evaluate the feasibility of adjunct guidance using on-line transesophageal echocardiography without intubation and general anesthesia in adolescent or adult patients undergoing transcatheter occlusion of secundum atrial septal defects. Ten consecutive cases (age 15-68 years) of secundum atrial septal defects with a pulmonary to systemic flow ratio of > 1.5 and a balloon-stretched diameter of < or = 25 mm were enrolled in the study. The Sideris' buttoned devices were used. The procedure was guided by on-line transesophageal echocardiography and fluoroscopy. Endotracheal intubation and general anesthesia were not employed. The diameters of the atrial septal defects ranged from 6 to 19 mm as determined by transesophageal echocardiography, from 7 to 20 mm as determined by atrial angiography and from 11 to 25 mm as determined by balloon sizing. The Sideris' buttoned devices were successfully deployed in all the patients. On-line transesophageal echocardiography greatly facilitated balloon sizing, device development and immediate assessment. One device unbuttoned 24 hours after the procedure and was retrieved smoothly. The remaining 9 patients were followed-up for 12 months. Two patients had no shunt, 3 had a trivial (average diameter by transesophageal echocardiography = 1-3 mm) and 4 had a small (average diameter = 4-6 mm) residual shunt at the latest follow-up. The cardiothoracic ratios decreased from 0.52 +/- 0.06 to 0.48 +/- 0.06 (p = 0.0131). There was no mortality, stroke or device fracture during the follow-up period. Thus, transcatheter occlusion of secundum atrial septal defect under adjunct guidance using on-line transesophageal echocardiography without endotracheal intubation and general anesthesia is promising for selected patients. PMID- 9850598 TI - The effects of starch and protein degradation rates, hay sources, and feeding frequency on rumen microbial fermentation in a continuous culture system. AB - Six continuous culture fermenters were used in three experiments to study the effects of dietary starch and protein degradability combination, hay sources, and feeding frequency on fermentation by rumen microorganisms. Experiments 1 and 2 used a 3 x 2 factorial design in which six diets were formulated to contain low (LS), medium (MS), or high starch degradability (HS) in combination with low (LP) or high protein degradability (HP). The dietary combinations were (1) LS + HP, (2) LS + LP, (3) MS + HP, (4) MS + LP, (5) HS + HP, and (6) HS + LP. In experiment 1, pangola was used as the hay source, and in experiment 2, alfalfa hay was used. In experiment 3, two starch degradabilities (LS, MS) and two hay sources (alfalfa, A; pangola, P) were combined with two feeding frequencies (2 X/d, 12 X/d). The dietary combinations were (1) LS + A + 12 X/d, (2) MS + A + 12 X/d, (3) LS + A + 2 X/d, (4) MS + A + 2 X/d, (5) LS + P + 12 X/d, and (6) MS + P + 12 X/d. A CRD design was used for experiment 3. Two rumen-cannulated Holstein cows fed alfalfa hay ad lib were used as donors of rumen fluid for all experiments. Each period was 6 d in length, with 5 d for adaptation and 1 d for sampling. In experiments 1 and 2, the effects of starch degradability on the composition of rumen microorganisms were significant. The MS or HS with HP had the highest total bacterial and protozoal density (P < 0.05). As for VFA, pH and nitrogen products, the effects of starch and protein degradability were not significant. There was no interaction between starch and protein degradability on most of microbial composition (ammonia-N, microbial nitrogen). In experiment 3, 12 X/d feeding frequency (MS + A + 12 X/d vs. MS + A + 12 X/d) resulted in higher pH, which tended to increase bacterial and protozoal density and starch and protein digestibilities. Different hay sources altered the starch and protein synchronization effect on the ammonia-N concentration (mg/dl) and non-ammonia N content (% DM) in the continuous culture system. PMID- 9850601 TI - Atmospheric deposition levels of chosen elements in the Czech Republic determined in the framework of the International Bryomonitoring Program 1995. AB - In order to determine the atmospheric loads of 13 elements (Al, As, Cd, Co, Cr, Cu, Fe, Mo, Ni, Pb, S, V, Zn), samples of Pleurozium schreberi (81.1%), Hypnum cupressiforme (11.2%) and Pseudoscleropodium purum (7.7%) bryophytes (mosses) were taken and analysed from an approx. 20 x 20-km grid extending over the entire territory (78,864 km2) of the Czech Republic (abbreviated 'the CZ' in this study). The level of the elements found in the bryophytes reflects the relative atmospheric deposition loads of the elements at the investigated sites. Five hot spots indicating relatively high deposition levels were identified in the CZ. The marginal hot spots are the following: the CZ part of the so-called Black Triangle I territory in northwestern CZ; the CZ part of the Black Triangle II territory in northeastern CZ; and the CZ part of the Sudeten mountains (Jizerske Mts and Giant Mts) and their foothills in northern CZ. Inland hot spots were found in the southwestern industrial part of central Bohemia and in the southern Moravian industrial district. The average element contents in CZ bryophytes were comparable with the respective average values obtained in Germany and Poland. However, the CZ average bryophyte values were higher and lower in comparison to the average Austrian and Slovak values, respectively. The CZ average relative atmospheric deposition loads of the elements were found to be 2-3 times higher than the respective loads in the cleanest parts of Europe (e.g. clean parts of Nordic countries). A comparison of the analytical results obtained repeatedly at 20 identical localities in the CZ showed a significant decrease in the relative deposition loads of all of the investigated elements in 1995 as compared to 1991. This decrease has been caused by the dramatic restriction of the industrial production, mainly that of the metallurgical and chemical industries, in the CZ. Desulphurisation programs and the effective trapping of flying dust particles in CZ power plants has also had a positive influence on the deposition climate in the CZ. The average values for the absolute atmospheric deposition of the investigated elements in the CZ in 1994-1995 as found from bryomonitoring are presented in this paper. PMID- 9850599 TI - cDNA cloning and expression analysis of the human UDPglucose dehydrogenase. AB - A cDNA clone encoding the human UDPglucose dehydrogenase was isolated from a liver cDNA library. The cDNA is 2,355 bp in length with an open reading frame which is capable of encoding a protein of 494 residues. The predicted primary sequence of the gene product is in good agreement with that of the bovine enzyme determined previously found by means of protein sequencing. Two major transcripts of the UDPglucose dehydrogenase gene with sizes of 2.8 and 2.35 kb, respectively, were observed by Northern analysis. The gene was found to be expressed in a variety of tissues with the highest level in liver, consistent with the physiological function of the enzyme in excretion of endo- and xenobiotics compounds. PMID- 9850600 TI - The geochemistry of mercury in central Amazonian soils developed on the Alter-do Chao formation of the lower Tapajos River Valley, Para state, Brazil. AB - In an oxisol-spodosol system developed on the terrestrial surface of the lower Tapajos Valley, the determination of total mercury (Hg), organic carbon (C), iron and aluminum oxy-hydroxide (Fe(cdb) and Al(cdb)) concentrations in the surface soil horizons are used to characterise the geochemical processes controlling the accumulation of Hg in soils under natural vegetation cover and in deforested and cultivated sites. Oxisols from the plateau have homogeneous and relatively high background Hg contents and burdens constituting an important natural reservoir of Hg for the region (90-210 ng/g dry wt. and 19-33 mg/m2 for the first 20 cm). The Fe(cdb) and Al(cdb) contents associated with the fine fraction (< 63 microns) of the soil suggest that oxy-hydroxides and, particularly Al-substituted Fe oxy hydroxides, control the Hg concentrations observed in all of the soils of the study region. Consequently, the geochemistry of these minerals along the slopes governs the accumulation or the release of the Hg according to the natural evolution of the soil cover and/or following the degradation of soils by erosion after deforestation and cultivation. These observations have important implications for the interpretation of Hg contamination patterns observed in Amazonian aquatic systems that could be linked to different drainage sources of Hg from the terrestrial surface. The sandification and podzolisation that is characteristic of the evolution of numerous pedological systems in the equatorial Amazon could be responsible for exportation of the naturally accumulated Hg, as for other metals, by acidic complexation and migration to the black waters of the Amazon. In the central Amazon region, as a result of the fragility of the soil cover, deforestation and cultivation, affecting principally the superficial soil, promote the selective erosion of fine particles enriched in oxides and Hg. The erosion of soil could be responsible for an important release of Hg, transported in particulate form by drainage waters. PMID- 9850603 TI - [Drainage and isolation of the work field in pediatric dentistry (II). A market overview and suitability study: the dam system and its use in pediatric dentistry]. PMID- 9850602 TI - [Drainage and isolation of the work field in pediatric dentistry (I). A market overview and suitability study: isolation of the work field without dams]. PMID- 9850604 TI - Principles of agonism: undressing efficacy. PMID- 9850605 TI - Nitric oxide: from discovery to the clinic. PMID- 9850606 TI - Ca2+ channel block and inactivation: common molecular determinants. PMID- 9850607 TI - The slope parameter and the receptor reserve. PMID- 9850608 TI - Does the noncompetitive NMDA receptor antagonist dizocilpine (MK801) really block behavioural sensitization associated with repeated drug administration? . AB - The term 'behavioural sensitization' refers to the progressively augmented behavioural response that is produced by many drugs of abuse upon their repeated administration. From most of the available data, it has been concluded that co administration of an N-methyl-D-aspartate (NMDA) receptor antagonist [such as dizocilpine (MK801)] together with the sensitizing drug can block the development of behavioural sensitization. However, the picture might not be that simple. Recent experimental evidence suggests that instead of blocking sensitization, co administration of dizocilpine enhances the effect of the the sensitizing drug or has more complex effects on the development of sensitization. In this article, Thomas Tzschentke and Werner Schmidt present these two different views and emphasize that the conclusions that can be drawn from sensitization experiments about the effects of dizocilpine and related drugs on behavioural plasticity crucially depend on how, when and under what conditions a test for sensitization is conducted. PMID- 9850609 TI - From Vanadis to Atropos: vanadium compounds as pharmacological tools in cell death signalling. AB - Vanadium compounds exert a variety of biological responses, the most notable being their effects as insulin mimetics. More recently, they have been used as pharmacological tools to investigate signalling pathways. Some peroxovanadium compounds act as powerful protein tyrosine phosphatase inhibitors, modulating both the extent and duration of phosphotyrosine signals at the level of the transmembrane growth factor receptors and targets in the cytoplasm and nucleus. A brief history of vanadium compounds, selected chemical properties of vanadium compounds and the ability of peroxovanadium complexes to modulate the activities of protein tyrosine phosphatases and tyrosine kinases are presented in this review by Anne Morinville, Dusica Maysinger and Alan Shaver. From the range of biological activities of these compounds, this review focuses on cytotoxic effects and possible roles of mitogen-activated protein kinases in mediating the effects exerted by vanadium compounds. PMID- 9850610 TI - Modulation of sensory nerve function in the airways. AB - Several clinical studies document a greater discrimination between asthmatic and healthy subjects in bronchial responsiveness to a range of stimuli such as cold air, distilled water and sodium metabisulphite, than to conventional bronchoconstrictor agonists including histamine and methacholine. One of the mechanisms thought to account for the bronchoconstriction induced by these agents is via reflex activation of the cholinergic pathway. An increase in sensory nerve (afferent) activity in asthma might account for the increased responsiveness to these agents. If so, a number of strategies are available to inhibit the function of afferent nerves which could lead to a suppression of bronchial hyperresponsiveness, including (1) inhibition of afferent activity, (2) inhibition of neuropeptide release and (3) antagonism of tachykinin receptors. As there are numerous reviews dealing with the latter, in this review Domenico Spina, Saloni Shah and Selena Harrison focus on the first two strategies. PMID- 9850611 TI - New developments in the molecular pharmacology of the myo-inositol 1,4,5 trisphosphate receptor. AB - Receptor-mediated activation of phospholipase C to generate inositol 1,4,5 trisphosphate [Ins(1,4,5)P3] is a ubiquitous signalling pathway in mammalian systems. A family of three IP3 receptor subtype monomers form functional tetramers, which act as effectors for Ins(1,4,5)P3, providing a ligand-gated channel that allows Ca2+ ions to move between cellular compartments. As IP3 receptors are located principally, although not exclusively, in the endoplasmic reticular membrane, Ins(1,4,5)P3 is considered to be a second messenger that mobilizes Ca2+ from intracellular stores. Ca2+ store mobilization by Ins(1,4,5)P3 can be shown to contribute to a variety of physiological and pathophysiological phenomena, and therefore the IP3 receptor represents a novel, potential pharmacological target. In this article, Rob Wilcox and colleagues review recent developments in IP3 receptor pharmacology, with particular emphasis on ligand molecular recognition by this receptor-channel complex. The potential for designing non-inositol phosphate-based agonists and antagonists is also discussed. PMID- 9850612 TI - [Chronic virus infections and development of cancer in genital and anal regions]. PMID- 9850613 TI - [Diagnostic imaging of thyroid gland diseases]. PMID- 9850614 TI - [Normocellular bacterial meningitis]. AB - Bacterial meningitis is usually associated with pleocytosis of the cerebrospinal fluid due to the presence of polymorphonuclear leucocytes. However, 75 cases of bacterial meningitis without pleocytosis have been published. Normocellular bacterial meningitis accounts for 3.5% (1-42%) of all patients suffering from bacterial meningitis and is seen in all age groups: The finding can be explained by three different mechanisms including 1) lumbar puncture performed early in the course of meningitis, 2) immune deficiency, and 3) relative leucopenia due to severe sepsis. Normocellular bacterial meningitis is in general associated with a good prognosis except for cases with severe underlying diseases. A high concentration of bacteria in a normocellular cerebrospinal fluid might also indicate a poor prognosis. PMID- 9850615 TI - [Sexually transmitted infection as a cause of anal cancer]. AB - Interviews were carried out with 423 women and 93 men with invasive or in situ anal cancer in Denmark and Sweden in a search for clues to the aetiology of this neoplasm. Patients with rectal adenocarcinoma (n = 534) and persons drawn from the background population (n = 554) served as controls. Multivariate logistic regression analyses confirmed previous observations of a strong association between either male homosexual experience or a history of anogenital warts and the risk for anal cancer. Moreover, hitherto unknown, but strong and consistent associations were observed between measures of high heterosexual activity and the risk for anal cancer among both sexes. Polymerase chain reaction analysis revealed human papilloma-virus DNA in the majority (88%) of anal cancer specimens but in none of 20 examined rectal adenocarcinomas. It is concluded that most anal cancers appear to be caused by sexually transmitted types of human papillomaviruses and, consequently, that anal cancer is a potentially preventable neoplasm. PMID- 9850616 TI - [Alcohol drinking patterns of the Danes 1979-1992]. AB - A questionnaire-based interview on alcohol drinking pattern in a random sample of 4,016 men and 4,179 women in 1992 was compared to an identical interview from 1979. In coherence with alcohol sale statistics, the average alcohol intake was unchanged in the population. Nevertheless, during the period drinking patterns have changed among Danes. Thus, 20 to 39 year-olds drank less, 40 to 59 years olds drank more, fewer Danes drank during daytime, and fewer drank on weekdays at the second survey. PMID- 9850617 TI - [Subjective symptoms and sleep apnea among persons referred to a sleep center. A questionnaire survey]. AB - The aim of this study was to evaluate the relation between symptoms and clinical characteristics and the frequency of increased apnoea and hypopnoea activity. The study population consisted of 135 persons referred to a sleep centre. One hundred and fifteen patients (85%) completed a questionnaire and were subsequently monitored during one night's sleep. Fifty-two persons (45%) had an Apnoea Hypopnoea Index (AHI = the mean number of apnoea and hypopnoea episodes per hour of sleep) > or = 10. Forty-one (36%) had an Apnoea Index (AI = the mean number of apnoea per hour of sleep) > or = 10. Symptoms and clinical characteristics had no significant correlation to increased apnoea activity; however, there was a significant relationship between self-reported frequent breathing pauses during sleep and increased AHI and AI. Symptoms and clinical characteristics are not capable of identifying persons with increased apnoea activity. PMID- 9850618 TI - [Can injuries cause by train crash be prevented? Experiences from the train crash in Jelling in 1995]. AB - Among 69 persons involved in a train crash, 53 were injured. Twenty-two persons were admitted to hospital. No-one was killed. There was a significantly increased risk of getting hurt when facing forward in the moving train. The mechanism of injury for the majority was being thrown forward and thereby hitting parts of the seats in front. The iron frames of the seats became exposed as some parts of the seats were only loosely attached. Arrangements to reduce injuries in train crashes could be an improved maintenance of the seats and their frames and the use of safety belts. PMID- 9850619 TI - [Hereditary antithrombin deficiency and pregnancy. Pregnancy course in six women with known hereditary antithrombin deficiency]. AB - Inherited antithrombin (AT) deficiency is a major cause of venous thromboembolism, especially in relation to surgery and pregnancy. We present six AT deficient pregnant women, who successfully delivered seven babies at the Department of Gynaecology/Obstetrics, Aalborg Hospital. From conception, or if possible prior to conception, the women were treated with unfractionated (UFH) or low molecular weight heparin (LMWH) throughout the pregnancy. If the pregnancy was without complication, AT substitution was only used at delivery and for approximately a week post-partum, when warfarin treatment was re-instituted. PMID- 9850620 TI - [Long-term oxygen therapy in chronic respiratory insufficiency]. AB - The aim of this retrospective study was to investigate the accordance between the indications used for prescription of continuous domiciliary oxygen and the guidelines recommended by the Danish Society of Pulmonary Medicine. Sixty-two patients with chronic hypoxic respiratory insufficiency in chronic domiciliary oxygen therapy were investigated. Only one patient fulfilled all the prescribed recommendations and, in general, there were no registrations of compliance or of follow-ups in the patient records. PMID- 9850621 TI - [Intestinal ganglioneuromatosis--a rare cause of chronic diarrhea]. AB - A case of intestinal ganglioneuromatosis is reported. The symptoms were watery diarrhoea and abdominal pain of several months duration. Endoscopic examination of the oesophagus, ventricle, duodenum, colon and rectum was normal. Mucosal biopsies from colon and rectum revealed ganglia cells and thin nerve fibres in the lamina mucosa, giving the diagnosis ganglioneuromatosis. As a consequence of the diagnosis thyroid scintigraphy, CT-scanning of the thyroid and adrenal glands and measurement of serum calcitonin and gastrin were performed. The tests revealed an intrathoracic nodular struma, and beyond this no abnormalities. The relation of intestinal ganglioneuromatosis to Multiple Endocrine Neoplasia type II b is discussed and the necessity of performing mucosal-biopsy from endoscopically normal colonic mucosa in cases of chronic diarrhoea is emphasised. PMID- 9850623 TI - [Hepatitis C-associated osteosclerosis. A rare disease with possible relevance for osteoporosis therapy]. PMID- 9850622 TI - [Prostacyclin inhalation in pulmonary failure following smoke inhalation]. AB - A 42 year-old man was admitted to hospital due to smoke inhalation. Thirty-three hours after admission arterial oxygen saturation was 80-90% with FiO2 = 1.0. Inhalation with prostacyclin was commenced with a dose rate of 7 ng/kg/min with an immediate effect, i.e. SATaO2 increased to 100%. Prostacyclin was terminated after 24 hours, when FiO2 was reduced to 0.5. It is concluded that prostacyclin inhalation therapy may have effect in patients following pulmonary smoke injury. PMID- 9850625 TI - [Can exercise be harmful?]. PMID- 9850624 TI - [Picture of the month. Air embolism]. PMID- 9850626 TI - [Should "Memento" be anonymous?]. PMID- 9850627 TI - Body height, body mass and surface area of the Neanderthals. AB - Body size, expressed as height or stature, is an important determinant of many other biological variables. Thus, it is surprising that many textbooks portray a wrong picture of Neanderthal height as being "very short" or "just over 5 feet". Based on 45 long bones from maximally 14 males and 7 females, Neanderthals' height averages between 164 and 168 (males) resp. 152 to 156 cm (females). This height is indeed 12-14 cm lower than the height of post-WWII Europeans, but compared to Europeans some 20,000 or 100 years ago, it is practically identical or even slightly higher. Considering the body build of Neanderthals, new body weight estimates show that they are only slightly above the cm/weight or the Body Mass Index of modern Americans or Canadians. The calculation of the relative surface area (approximately 240-244 cm2/kg) is very low and supports earlier findings of a morphological and anatomical thermoregulatory adaptation to a cold climate in the Neanderthals. PMID- 9850628 TI - Facial reconstructions on the Vors-Papkert B Cemetery series. AB - Seven skulls from the Vors-Papkert B Cemetery were analysed according to the facial reconstruction method of KOLLMANN and GERASIMOV. The degree of expression of sex, metric data of the skull, morphological characteristics of the skull and characteristics of the reconstructed face are presented for each of the skulls. PMID- 9850629 TI - Patterns of faciodental sexual dimorphism in Hispanopithecus. AB - This paper examines patterns of sexual dimorphism within the Spanish hominid Hispanopithecus. The inclusion of this genus within the Hominidae suggests that extant male and female great ape sexually specific characters are appropriate features for determining the sex of these fossil hominids. It is shown that Hispanopithecus has a distinct sexually dimorphic pattern from that observed in the extant great apes. It is also demonstrated that H. laietanus and H. crusafonti each have a distinct sex pattern from each other. This result further supports the dual species hypothesis. PMID- 9850630 TI - [Is head size modified by environmental factors?]. AB - In the article the development of skull measurements and head measurements (length and breadth) and of the cephalic index, calculated from these measurements, since the Neolithic period are presented. The results obtained from the historical material are compared with those of living persons. The measurements as well of the skull as of the head show secular changes. The following general trend was found: an increase of body height is connected with a debrachycephalisation and a decrease of body height is connected with a brachycephalisation. It can be emphasized that brachycephalisation/debrachycephalisation are part of the secular trend. Therefore environmental factors are responsible for the described changes of measurements of the skull and the head in a broadest sense. PMID- 9850631 TI - Size comparison of the male and female human corpus callosum from autopsy samples. AB - 161 cases of human-brain autopsies (93 Male, 68 Female) were collected from three samples in the Kinai district of Japan. From photographs of the midsagittal plane of each brain, surface areas of the corpus callosum (CC in mm2) and of the medial view of the cerebrum (H in mm2) were measured. The absolute H and CC values from each of the three samples were larger in males than in females, with full overlapping of the male and female ranges for both H and CC values. However, the relative values of CC corrected for H (CC/H) were almost equal between sexes. The relative values for the posterior one-fifth (splenium) of the CC, which were divided by CC or H, were slightly higher in females than in males in each of the three samples, but without statistical significance. This trend was also found in the relative values for the posterior one-third of the CC and in the relative values for region, meaning the posterior one-fifth of the CC subtracted from the posterior one-third of the CC (isthmus), but not in the relative values for the anterior one-fifth (genu) of the CC. PMID- 9850632 TI - Formation of morphological traits of the quadriceps femoris muscle (QFM) before and after birth. Part IV. Vastus intermedius muscle (VIM). PMID- 9850633 TI - Longitudinal analysis of adolescent growth of foot length and stature of children living in Ogi area of Japan: a 12 years data. AB - Adolescent growth of foot length (FL) and stature were investigated in a sample of 586 children living in the Ogi area of Saga Prefecture Japan, who were traced longitudinally from 6.5 to 17.5 years of age. FL growth and its relationship to height were analyzed using the JPA2 model and the Optimal Kernel Method to fit the serial data of stature and FL. The results demonstrated that there was clear peak growth velocity (PGV) during the adolescent growth period of FL for most of the children; before 11.5 years of age there was no essential difference in mean foot length between girls and boys; the completion of FL growth was on average two years earlier than that of stature and two and a half years after the appearance of the peak height velocity (PHV); girls completed the FL growth two years earlier than boys; the longer the growth period for FL or the larger the value of FL at an early age the bigger the adult foot length; the foot grew in synchrony with the body as a whole. PMID- 9850634 TI - Prediction of maximal aerobic power in healthy Indian males 21-58 years of age. AB - An attempt has been made in this paper to predict maximal aerobic power (VO2max) in healthy Indian males by three important physical characteristics (viz. age, height and body weight) using the technique of multivariate regression analysis for a wide age group ranging from 21 to 58 years. It has been observed that these characteristics can contribute significantly to the prediction of maximal aerobic power. Three combinations of predictors of two predictors at a time have also been tried to predict maximal aerobic power. Out of these three combinations, the combination of age and body weight has the maximum multiple correlation (R = 0.536, P < 0.001) for a group of 146 subjects of healthy Indian males. A nomogram has been constructed to predict VO2max from age and body weight in order to make the developed multiple linear regression equation of more practical utility to the biomedical scientists who may not be well acquainted with the statistical computational work. PMID- 9850635 TI - New diagnostic techniques applicable to fine needle aspirates. PMID- 9850636 TI - Reverse transcriptase polymerase chain reaction on fine needle aspirates for rapid detection of translocations in synovial sarcoma. AB - OBJECTIVE: To evaluate the utilization of fine needle aspiration (FNA) biopsy to obtain material for reverse-transcriptase polymerase chain reaction (RT-PCR) in the detection of the t(X;18)(p11.2;q11.2) translocation in synovial sarcomas. STUDY DESIGN: We applied RT-PCR to detection of synovial sarcoma fusion gene transcripts on fine needle aspirates. Five clinical samples were first analyzed: one was a tumor previously diagnosed as malignant hemangiopericytoma, one was a poorly defined tumor, and three were suspected synovial sarcomas. FNA material was transferred directly to the RT-PCR reaction tube without RNA extraction. RESULTS: The t(X;18) translocation could be detected on the limited amount of material that FNA provides. In each of the cases studied the representivity of the tumor samples was confirmed microscopically. CONCLUSION: Our protocol permits analysis directly on representative samples without extraction of RNA. The results imply that RT-PCR offers reliable detection of sarcoma fusion gene transcripts on fine needle aspirates. The procedure, apart from being applicable to outpatients, is rapid and sensitive. PMID- 9850637 TI - Correlation of fine needle aspiration biopsy and fluoride-18 fluorodeoxyglucose positron emission tomography in the assessment of locally recurrent and metastatic head and neck neoplasia. AB - OBJECTIVE: Patients with primary head and neck neoplasia can present during follow-up with suspected recurrence, and both fine needle aspiration biopsy (FNAB) and fluoride-18 fluorodeoxyglucose positron emission tomography (FDG-PET) scan are available methodologies for evaluating these patients. Our objective was to retrospectively correlate patients who underwent both FNAB and FDG-PET scan in order to assess the possibility of recurrent neoplasia. STUDY DESIGN: The cytopathology files at Saint Louis University Health Sciences Center were retrospectively searched for patients with known primary head and neck malignancies beginning in 1995. Suspected recurrence and local metastases evaluated by both FNAB and FDG-PET scan were correlated. RESULTS: Twenty-eight patients received a combined total of 37 FNABs with concurrent FDG-PET scans. The majority of patients had primary oropharyngeal squamous cell carcinoma with intermixed, single cases of other primary head and neck neoplasms. Thirty of the 32 aspirates with recurrent or locally metastatic disease had combined positive findings by both FNAB and FDG-PET scan, yielding a sensitivity of 94%. One nonspecific and one negative FDG-PET scan came from a patient who had disease confirmed by FNAB. Five patients had negative findings by both methods that were supported by the subsequent clinical course. CONCLUSION: FNAB can provide confirmatory evidence of disease in a clinically suspicious abnormality with nonspecific FDG-PET results. FNAB and FDG-PET are highly sensitive for tumors in cases of clinically suspected recurrence and locally metastatic disease. PMID- 9850638 TI - Improved detection of adenocarcinoma of serous fluids with p53 immunocytochemistry. AB - OBJECTIVE: To investigate the potential value of p53 protein immunostaining in identifying malignant cells in serous fluids. STUDY DESIGN: We applied p53 immunostaining to 26 cytologically malignant, 8 suspicious and 34 benign specimens of serous fluids from 68 patients. For comparison, staining for carcinoembryonic antigen (CEA) was also done on all the specimens. RESULTS: CEA was positive in 23 of 26 (88%) cytomorphologically malignant, 3 of 8 (38%) suspicious and 1 of 34 benign cases. p53 Nuclear immunostaining was positive in 12/26 (46%) malignant, 2/8 (26%) suspicious and no benign cases. Correlation between p53 staining and serous fluid type (benign, suspicious or malignant) was significant. The P based on Fisher's exact test was < .0001. Two cases that were reported cytomorphologically as suspicious stained positively with p53; further investigation in those cases confirmed the diagnosis of metastatic adenocarcinoma. CONCLUSION: p53 Immunostaining of serous fluids seems to be of value in identifying carcinoma cells, especially in those cases that show inconclusive or bland cytologic features. Combining p53 with CEA immunostains in clinically or cytologically suspicious cases may assist in recognition of carcinoma cells and in pursuing an appropriate therapeutic approach. PMID- 9850639 TI - Fine needle aspiration cytology of penile tumors. AB - OBJECTIVE: Penile tumors are unusual and can represent neoplastic or inflammatory processes. We report a series of penile tumors diagnosed by fine needle aspiration (FNA) cytology. STUDY DESIGN: Files from the Division of Clinical Cytology, Karolinska Hospital, were searched, and cases of FNA cytology of the penis selected. Clinical information and cytologic diagnoses were recorded and observations from aspiration smears made. RESULTS: Eleven patients had presented to Karolinska Hospital for FNA cytology of the penis. They ranged from 46 to 89 years of age; all were uncircumcised. Tumors were present in various locations, including the glans and shaft. Five patients had squamous cell carcinoma (SCCA) of the penis; three were primary, one locally recurrent and one metastatic from primary lung SCCA. There were three carcinomas metastatic from primary colon, prostate and bladder tumors. There was one each of locally recurrent malignant melanoma, locally metastatic testicular mesothelioma and Peyronie's disease. Discomfort associated with the procedure was tolerable, there were no complications, and each biopsy provided sufficient material for a cytologic diagnosis. CONCLUSION: FNA cytology of penile tumors is a successful, well tolerated procedure capable of providing a cytologic diagnosis and useful information for patient management. PMID- 9850640 TI - Fine needle aspiration cytology of dermatopathic lymphadenitis. AB - OBJECTIVE: Dermatopathic lymphadenitis (DL) can be a nonneoplastic cause of an enlarged lymph node or nodes. Cytologic findings of DL have not been well described. The differential diagnosis includes Langerhans histiocytosis (LH) in children and low grade lymphoma in adults. We present three cases of dermatopathic lymphadenitis with a discussion of cytologic findings and differential diagnosis. STUDY DESIGN: Fine needle aspiration (FNA) findings of three cases of lymph nodes involved by DL were reviewed. All three were axillary lymph nodes in patients with skin rashes. Immunoperoxidase stains for S-100 and CD1a were performed on a cell block from one case, and flow cytometric analysis was performed on another. RESULTS: FNA of DL yields cellular smears with abundant histiocytoid cells with moderate cytoplasm and ovoid, vesicular nuclei with longitudinal grooves (interdigitating reticulum cells). These histiocytoid cells are S-100 and CD1a positive. The background cells are mature lymphocytes, scattered eosinophils and plasma cells. Case 1 was originally misinterpreted as consistent with LH. In case 2, flow cytometric analysis of a mixed population of lymphocytes was seen, ruling out lymphoma. In case 3, the diagnosis of DL was based on morphologic features. CONCLUSION: Interdigitating reticulum cells in dermatopathic lymphadenitis are S-100 and CD1a positive and could be confused with LH in children. In adults, flow cytometry could be used to distinguish them from low grade lymphoma. FNA findings of DL should be interpreted with the clinical history. PMID- 9850641 TI - Fine needle aspiration cytology of dermatopathic lymphadenitis. AB - OBJECTIVE: To delineate and describe the cytomorphologic features of dermatopathic lymphadenitis on fine needle aspirated material. STUDY DESIGN: Thirteen cases of previously diagnosed dermatopathic lymphadenitis (DLN) and 10 cases of reactive lymphadenitis (RLN) (5 each of sinus histiocytosis and follicular hyperplasia) were reexamined by two independent observers. The cytologic features were semiquantitated (0-3+). Histology was available in two cases. RESULTS: A variable amount of melanin pigment was seen in all 13 cases of DLN and in only 3 cases of RLN. DLN had large histiocytic collections with blood vessels at the center (12/13). The histiocytic cells had an elongated nucleus (13/13) with nuclear grooves (13/13) and pseudonucleoli (11/13). Many of these nuclei had a convoluted and crumpled appearance (11/13). On immunocytochemistry these cells were S-100 positive (six cases) and CD68 negative (one case). In contrast, in RLN with sinus histiocytosis, the histiocytes were polygonal and had rounded nuclei. Spindle-shaped histiocytes were rare, while pseudonucleoli and nuclear grooves were absent. Sinus histiocytes were CD68 positive and S-100 negative on histologic sections. Tingible body macrophages were rare (2/13) in DLN and prominent (8/10) in RLN. Mitoses were seen in DLN (6/13) and RLN (9/10). CONCLUSION: The morphologic features found to be helpful in the diagnosis of DLN on fine needle aspiration cytology are melanin-laden macrophages with variable pigment; large, histiocytic clusters with blood vessels at the center; characteristic histiocytes, with elongated vesicular nuclei, nuclear grooves, crumpled and convoluted nuclei and pseudonucleoli; and absence of or very few tingible body macrophages. Positivity on immunostaining for S-100 and negativity for CD68 aid in the diagnosis. PMID- 9850642 TI - Fine needle aspiration cytology of benign adrenal cortical nodules. A comparison of cytologic findings with those of primary and metastatic adrenal malignancies. AB - OBJECTIVE: To determine the specific cytomorphologic criteria for diagnosing benign adrenal cortical nodule (BACN) by fine needle aspiration (FNA). STUDY DESIGN: The smears from 162 adrenal FNA biopsies were reviewed. A diagnosis of BACN was rendered in 50 cases (31%). The cytologic features of BACN were compared to those of primary and metastatic malignant tumors of the adrenal gland, and the size of BACN as measured by computed tomography was recorded. RESULTS: Bubbly, vacuolated, lipid background; large, cohesive tissue fragments with a syncytial nesting arrangement admixed with sinusoidal endothelial cells; and abundant oval, round, bare nuclei are the three cytomorphologic features most often seen in BACN. The combination of these three features was observed in 40 cases (89%) of BACN and was seen in 4 cases of metastatic carcinoma (6%) in which there was also coexisting adrenal cortical hyperplasia. None of the other primary or metastatic malignancies showed this combination of cytomorphologic features. The mean size of BACN was 2.5 cm, with a standard deviation of 1 cm and a range of 1-5 cm. The sizes of the four metastatic carcinomas with coexisting adrenal cortical hyperplasia were 6.5, 6, 5 and 1.5 cm, respectively (mean, 4.8). CONCLUSION: Our data suggest that combined cytologic features of bare nuclei; bubbly, vacuolated background; and large, cohesive tissue fragments with sinusoidal endothelial cells in a small adrenal nodule (< 3.5 cm) are highly specific to FNA diagnosis of BACN. PMID- 9850643 TI - Fine needle aspiration cytodiagnosis of liver tumors. Results obtained with Riu's stain. AB - OBJECTIVE: To present our experience with liver fine needle aspiration (FNA) diagnosis based on Riu's stain. STUDY DESIGN: We reviewed a total of 322 liver fine needle aspirates from 286 patients seen in a seven-year period from April 1990 to April 1997 at Koo Foundation Sun Yat-Sen Cancer Center, Taipei. Surgical and/or clinical follow-up was available for confirmation in 292 aspirates. RESULTS: The cytologic diagnosis was categorized into four groups: benign in 81 cases, suspicious in 13, malignant in 225, and inadequate specimen in 3 cases. There were 16 false negative and no false positive diagnoses. Two suspicious aspirates were negative. Our results showed a sensitivity of 93.3% and a specificity of 100% for the detection of malignancy. If suspicious cases were considered positive, the specificity decreased to 95.1%, while the sensitivity increased to 93.6%. Among 87 hepatocellular carcinomas (HCCs) in our series, correct FNA diagnosis was made in 84 cases with an accuracy of 96.6%. Out of 135 cases of non-HCCs, 1 was incorrectly diagnosed. The accuracy of identifying a liver malignancy as non-HCC was 99.3%. CONCLUSION: Cytologic features of HCC are well demonstrated by Riu's stain, with high accuracy in identifying them. Liver FNAs using Riu's stain combined with cell block study and clinicopathologic correlation can achieve very high sensitivity and specificity in the detection of hepatic malignancies. PMID- 9850644 TI - Clinical significance of "cannibalism" in urinary cytology of bladder cancer. AB - OBJECTIVE: To investigate the cytologic characteristics and clinical significance of "cannibalism" (cytophagocytosis), a characteristic histologic finding associated with malignancy, found in urinary cytology samples during the diagnosis of bladder cancer. STUDY DESIGN: The subjects were 252 patients with bladder cancer initially treated from 1981 to 1991. For their pretreatment urine samples, membrane filtration was performed to determine the presence and amount of cannibalism, a tumor cell within a tumor cell. The urinary cytologic findings were then correlated with the histologic findings of primary bladder cancer. RESULTS: Only cells from urinary cytology-positive specimens demonstrated cannibalism. The positive rate of cannibalism was significantly higher in grade 3 (25%) than grade 1 (0%) or 2 (8%) superficial papillary cancer (P < .01) and also higher in muscle-invading bladder cancer (57%) than grade 3 superficial papillary cancer (P < .01). The rate did not differ significantly between muscle-invading cancer and superficial nonpapillary cancer (44%). In 198 patients with superficial bladder cancer, progression was observed in 19 (10%) during a mean follow-up of 72 months. All of them showed positive urinary cytology results in pretreatment samples, and among the patients with positive urinary cytology, the progression rate was significantly higher when cannibalism was present than when it was absent (38% vs. 17%, P < .05). Moreover, in high grade cases, cannibalism had significant predictive value for progression. By multivariate analysis, cannibalism was an independent factor for prediction of progression, as were tumor grade and stage. CONCLUSION: Cannibalism in urinary cytology appears to be an indicator of both the anaplastic grade and invasiveness of transitional cell carcinoma of the bladder. Furthermore, cannibalism may provide a reliable predictor of progression of superficial bladder cancer. PMID- 9850645 TI - Reliability and accuracy in reporting CIN in 14 laboratories. Developing new indices of diagnostic variability in an interlaboratory study. The Working Group for External Quality Control in Cervical Cytopathology. AB - OBJECTIVE: To report the results of the second phase of a pilot study of an Italian national external quality evaluation program (EQA) aimed at assessing whether participation in the first phase had increased the reliability and accuracy of diagnoses and to what extent. STUDY DESIGN: In the second phase, two sets of cervical smears (similar to the ones examined in the first phase) were circulated among the 14 participating cytologic laboratories throughout Italy. Responses were recorded on a standardized form. Participants were asked to judge the adequacy of each smear and to formulate a diagnosis. They were also asked to recommend management of the patient on the basis of the smear report and to evaluate the degree of diagnostic difficulty of each slide. The results were discussed in workshops, and it was possible to reach a consensus diagnosis on 37 of 40 smears. In the statistical analysis, new indices of diagnostic variability were developed and calculated; a gross index of agreement, unweighted and weighted kappas, analysis of exchangeability, sensitivity and specificity were also estimated. RESULTS: The results of the second phase are similar to those of the first phase and no substantial improvement in accuracy and little reduction in variability were observed. The interventions carried out in this study (discussion between representatives of laboratories of diagnostic differences and reassessment of the most controversial slides) were aimed at increasing consensus among the participating pathologists but were insufficient to change the diagnostic routines in their laboratories. CONCLUSION: It may be advisable to promote two kinds of interlaboratory quality programs with two separate but integrated components: (1) a core component with slides having clear-cut diagnoses, and (2) a continuing education component. PMID- 9850646 TI - Prognostic features of cervical dysplasia associated with specific types of HPV DNA and cytologic features characteristic of HPV infection in dysplasia. AB - OBJECTIVE: To study a possible etiologic relationship between the prognosis of uterine cervical dysplasia, association of type-specific human papillomavirus (HPV) DNA and cytologic features characteristic of HPV infection. STUDY DESIGN: Two hundred thirteen cases of uterine cervical dysplasia were selected in which follow-up survey for more than two years was possible. Frequency of the presence of HPV DNA in the DNA samples was determined by polymerase chain reaction. The cervical scrapings were also examined microscopically for the frequencies of cells with cytologic features characteristic of HPV infection. RESULTS: HPV was positive in 98 cases (46.0%). The high-risk type of HPV was detected at almost the same frequencies in both progressive and regressive states of dysplasia. Cytologic features were more evident in cells infected with the low-risk type of HPV. CONCLUSION: Involvement of an as-yet-unknown factor or factors coupled with infection with the high-risk type of HPV is implicated in the progression of uterine cervical dysplasia. Cytologic features characteristic of HPV infection may serve as a diagnostic marker for a favorable prognosis in dysplasia. PMID- 9850648 TI - Salivary duct carcinoma. Is a specific diagnosis possible by fine needle aspiration cytology? AB - OBJECTIVE: To review the cytomorphologic features of salivary duct carcinoma and to evaluate the likelihood of definitive diagnosis by fine needle aspiration. STUDY DESIGN: The cytomorphologic features of two cases of salivary duct carcinoma, both occurring in the parotid gland in men over 80 years of age, were evaluated by fine needle aspiration and compared to cytologic features described in the literature. Additionally, previously reported diagnoses rendered by fine needle aspiration of salivary duct carcinomas were compiled from the cytology literature. The likelihood of arriving at a definitive diagnosis by fine needle aspiration was determined from the frequency of correct cytologic diagnoses reported in the literature. RESULTS: The most characteristic features of salivary duct carcinoma by fine needle aspiration appear to be flat sheets with a cribriform pattern and tumor cells in a necrotic background with pleomorphic, eccentric nuclei and granular cytoplasm. However, no definitive diagnoses of salivary duct carcinoma by fine needle aspiration have been recorded in the cytology literature. CONCLUSION: Because of the morphologic spectrum displayed by this tumor and the absence of definitive cytologic diagnoses in the literature to date, it is unclear whether a diagnosis of salivary duct carcinoma can be rendered by fine needle aspiration. Nevertheless, if cribriform groups are noted in a salivary gland aspirate, the diagnosis of salivary duct carcinoma should at least be considered. PMID- 9850647 TI - Thyroid transcription factor-1 and cytokeratins 7 and 20 in pulmonary and breast carcinoma. AB - OBJECTIVE: To evaluate the immunohistochemical expression of a lung epithelial gene transcription factor, thyroid transcription factor-1 (TTF-1), in lung and breast carcinoma in pulmonary cytologic preparations and to correlate the results with the expression of cytokeratin 7 (CK7) and 20 (CK20). STUDY DESIGN: Cell blocks of cytologic specimens were immunostained with antibodies to TTF-1, CK7 and CK20. Specimens included 41 primary lung carcinomas (21 adenocarcinomas, 8 squamous cell carcinomas and 12 small cell undifferentiated carcinomas) and 6 metastatic breast adenocarcinomas. RESULTS: The lung adenocarcinomas showed nuclear reactivity for TTF-1 in 76% (16/21) of the cases and a staining combination of CK7+/CK20- in 95% (20/21) of the cases. Only one case was CK7+/CK20+. All the breast carcinomas were nonreactive to TTF-1, and all were CK7+/CK20-. The squamous cell carcinomas and small cell undifferentiated carcinomas showed TTF-1 positivity in 38% (3/8) and 83% (10/12), respectively. PMID- 9850649 TI - Glutathione S-transferase pi immunostaining of cisplatin-resistant ovarian cancer cells in ascites. AB - OBJECTIVE: To clarify immunohistochemically the correlation between glutathione S transferase pi (GST pi) expression of surgically resected specimens and clinical response in ovarian cancer and to evaluate ascites cytology using GST pi staining. STUDY DESIGN: Eighty-seven patients with ovarian cancer underwent initial debulking surgery and received cisplatin-based chemotherapy after surgery. Immunostaining for GST pi was performed on formalin-fixed sections of the patients' tumors. The cytologic slides of 24 cases were acquired for evaluation of GST pi staining. RESULTS: Of 87 surgically resected specimens, 55 (63.2%) were GST pi positive. Twenty-five of 28 patients (89.3%) who showed no response to chemotherapy had GST pi-positive tumor cells. The predictive value of positive GST pi staining for drug resistance was 75.8% (25/33). Of 18 cases that were GST pi positive in surgically resected specimens, 17 were positive in ascites cytology. Five cases were negative in both resected specimens and ascites cytology. There was a significant correlation in the GST pi labelling index between resected specimen and ascites cytology from the same case; the correlation coefficient was .701 and P value < .001. CONCLUSION: Overexpression of GST pi is related to resistance to cisplatin, and GST pi staining of ascites cytology can be used in pretherapeutic assessment of patients with ovarian cancer. PMID- 9850650 TI - Benign glandular cells in posthysterectomy vaginal smears. AB - OBJECTIVE: To investigate the occurrence and origin of benign glandular cells in posthysterectomy vaginal smears and to propose a categorization for these cells under the Bethesda system. STUDY DESIGN: Among 4,986 posthysterectomy vaginal smears during a 3.5-year period, 82 patients were identified with smears containing benign glandular cells. The review of the smears, related biopsies and charts form the basis of this report. A control group of 236 posthysterectomy smears without glandular cells from the same period was reviewed. RESULTS: Smears were available for review on 76 of 82 patients. All were within normal limits or showed benign cellular changes. All contained clearly benign glandular cells, most frequently present in groups. In all smears, pale pink-red intracellular mucin was identified either diffusely within the cytoplasm or within vacuoles. The glandular cells appeared as hybrid parabasal-endocervical type, squamous metaplastic or columnar cells. Associated cytologic findings included atrophy, inflammation, blood and repair. Reactive atypia was present in 10 (13%) cases. Biopsies were performed on 40 (48%), all showing benign changes. In four of five cases stained, intracellular mucin was seen within glandular cells. Chart review revealed that the patients' hysterectomies were performed for removal of a malignant tumor in 58 (71%) of the study group and in 100 (42%) in the comparison group and that 48 (58%) of the study group had received radiotherapy and/or chemotherapy, while only 26 (11%) of those in the comparison group had similar therapy. CONCLUSION: The presence of benign glandular cells in posthysterectomy vaginal smears is an infrequent but not rare finding. This study indicated an association with benign processes and showed a frequent association with prior radiation or chemotherapy. A more appropriate designation under the Bethesda System is "benign cellular changes" rather than "glandular cell abnormalities." An aggressive workup does not appear to be warranted in this group of patients. We speculate that this phenomenon represents a metaplastic process, possibly secondary to radiation or chemotherapy. PMID- 9850651 TI - Kawan bronchial brush/cell block technique. Facilitation of the routine diagnosis of bronchial neoplasms. AB - OBJECTIVE: To evaluate cell block preparations as a tool for the diagnosis of pulmonary malignancy and to correlate the findings with those from brush cytology and histology from forceps biopsy. STUDY DESIGN: One hundred twenty consecutive samples from patients with primary or metastatic lung cancer were routinely processed and stained for cytologic and histologic examination. In addition to smears, a cell block was prepared from each brushing sample. Brush smears, cell blocks and biopsy specimens were compared and evaluated for their diagnostic accuracy. RESULTS: Brush cytology samples were interpreted as positive for malignancy in all 120 cases. In 42 cases immunohistochemistry performed on cell blocks led to the final precise type diagnosis. Owing to technical problems (stenosis, bleeding, peripheral location), forceps biopsy specimens were obtained from only 51 patients. CONCLUSION: In addition to lower patient risk, cell block preparation yields high diagnostic accuracy and may thus be considered an improvement in quality assurance. PMID- 9850652 TI - Comparison of aspiration, touch and scrape preparations simultaneously obtained from surgically excised specimens. Effect of different methods of smear preparation on interpretive cytologic features. AB - OBJECTIVE: To determine whether three commonly used methods of obtaining and preparing cells for cytologic evaluation--touch preparation (TP), fine needle aspiration (FNA) and scrape preparation--yield essentially comparable material for evaluation or whether important differences exist between them that may influence interpretation. STUDY DESIGN: FNA, TP and scrape preparations were made simultaneously on surgically excised specimens. Thirty breast, 6 lung and 12 colon lesions were studied. Each slide was assessed for qualitative and quantitative features. Cellularity was semiquantitatively scored. RESULTS: Scrape preparations were the most cellular (P < .0001). Scrape and TPs often had larger tissue fragments than FNA. However, fragmentation of epithelial groups into smaller clusters and single cells was often noted on scrape preparations. FNAs tended to have cleaner backgrounds than the other two methods. CONCLUSION: Most often, there are few differences between smears prepared by the three techniques studied. However, scrape preparations may yield smears with smaller clusters and single cells as compared to the other two methods; that is a potential source of diagnostic confusion, particularly with respect to benign breast lesions. Scrape preparations uniformly yield more cellular smears. To the extent that cellularity is an interpretive factor in assessing cytologic specimens, it is important to be aware of the increased cellularity of scrape preparations as compared to the other two techniques. PMID- 9850653 TI - Diagnosis of renal malacoplakia by fine needle aspiration cytology. A case report. AB - BACKGROUND: Malacoplakia is a rare chronic inflammatory disease first described by von Hansemann in 1901. L. Michaelis and M. Gutmann published the first report in 1902, describing the characteristic round intracellular and extracellular inclusions, or Michaelis-Gutmann bodies. Renal malacoplakia is a form of chronic tubulointerstitial nephritis the clinical and radiologic findings of which are often suggestive of a neoplasm. Renal malacoplakia may be related to megalocytic interstitial nephritis and xanthogranulomatous pyelonephritis. CASE: A 67-year old male presented with acute left loin pain suggestive of uretery colic. The past medical history was unremarkable and physical examination within normal limits. Renal ultrasound and computed tomography showed a 3.5-cm, cystic mass with a thickened, irregular wall located centrally within the left renal sinus. The size of the left kidney was 11.5 cm. A renal arteriogram provided no evidence of a vascular mass of any type. Fine needle aspiration (FNA) revealed many large, foamy, granular macrophages with large, eccentric nuclei and prominent nucleoli containing round inclusions that exhibited a laminated appearance, consistent with the structure of Michaelis-Gutmann bodies. Many were also present extracellularly. CONCLUSION: To our knowledge this is the first case of renal malacoplakia diagnosed by fine needle aspiration. This case demonstrates the value of FNA in determining the nature of a radiologically nonspecific lesion. It highlights the value of FNA as a substitute for or preliminary method before more invasive procedures in the diagnosis of a treatable disease entity. PMID- 9850654 TI - Cytology of primary pulmonary meningioma. Report of the first multiple case. AB - BACKGROUND: Ectopic meningiomas arising in the lung are rare. We report here the first multiple primary case diagnosed by intraoperative imprint cytology. CASE: Asymptomatic pulmonary nodules, two in the left and three in the right lung, were found in a 61-year-old woman, and video-assisted thoracoscopic surgery was undertaken. Because the largest tumor was diagnosed as a meningioma by intraoperative imprint cytology using an excised biopsy specimen, further resection was not performed immediately. Histopathologically the tumor was characterized by whorled nests of cells accompanied by psammoma bodies intermingled with a fibrous pattern. The diagnosis was a transitional meningioma, positive for vimentin and epithelial membrane antigen and negative for keratin immunohistochemically. All the nodules were subsequently surgically resected and showed a similar cytohistologic appearance. Ultrastructurally the tumor cells demonstrated interdigitation of adjacent plasma membranes with numerous desmosomes and hemidesmosomes, typical of meningiomas. We failed to detect another primary tumor in the nervous system, and at this writing the patient was healthy three years after the operation. CONCLUSION: Because of the characteristic cytomorphologic features of primary pulmonary meningioma, the cytologic approach provides useful information for therapy. PMID- 9850655 TI - Submandibular gland carcinoma ex pleomorphic adenoma. Report of a case with cytologic features and diagnostic pitfalls. AB - BACKGROUND: Carcinoma ex pleomorphic adenoma is a rare neoplasm of the salivary gland. This lesion, also known as malignant mixed tumor, occurs when a malignant tumor arises in the epithelial component of a pleomorphic adenoma. Reports of fine needle aspiration biopsy (FNAB) diagnosis of malignant mixed tumors are rare and have been limited to cases arising in the parotid. Cytologic features and diagnostic pitfalls of this uncommon neoplasm are presented. CASE: A 75-year-old male presented with a nontender submandibular mass. The lesion had been present 12 months, with a recent increase in size. FNAB was performed, and the smears revealed a mixture of benign and malignant areas. The benign portion of the smears showed findings typical of pleomorphic adenoma. The malignant area showed large cells occurring singly and in groups. The malignant cells contained pleomorphic nuclei with irregular nuclear membranes and prominent macronucleoli; cytologically, they resembled cells from a poorly differentiated adenocarcinoma. CONCLUSION: We present the first case of carcinoma ex pleomorphic adenoma of the submandibular gland correctly diagnosed by FNAB. This rare salivary gland malignancy can be accurately diagnosed on FNAB if strict criteria are applied. PMID- 9850656 TI - Papillary Hurthle cell carcinoma (Warthin-like tumor) of the thyroid. Report of a case with fine needle aspiration findings. AB - BACKGROUND: Papillary Hurthle cell carcinoma with lymphoplasmacytic stroma is a newly recognized variant of papillary carcinoma of the thyroid gland. Fine needle aspiration findings reveal some points that have not been reported previously. CASE: A 50-year-old female presented with a left lobe thyroid mass. Fine needle aspiration of the nodule yielded cellular smears, mostly in large and small papillary clusters of Hurthle cells, with nuclear grooves and rare nuclear inclusions in a lymphoplasmacytic background. Isolated Hurthle cell and multinucleated giant cells were also seen. Histopathologic examination of the 3 cm-diameter nodule showed papillary Hurthle cell carcinoma with lymphocytic stroma, or "Warthin-like tumor" of the thyroid, with capsular invasion. CONCLUSION: The presence of papillary clusters with nuclear features of papillary carcinoma and oxyphilic cytoplasm in a lymphoplasmacytic background should raise the diagnosis of Warthin-like tumor of the thyroid. PMID- 9850657 TI - Macrofollicular variant of papillary thyroid carcinoma. Report of a case with fine needle aspiration biopsy findings. AB - BACKGROUND: Macrofollicular variant of papillary thyroid carcinoma is defined as a combination of numerous macrofollicles occupying 50% of the tumor and foci of the conventional follicular variant of papillary carcinoma. To the best of our knowledge, there have been no English-language reports on its cytologic findings. CASE: A fine needle aspiration specimen from a mass of the thyroid in a 59-year old female showed a large collection of dense, inspissated colloid and a few sheets of tumor cells with nuclei characteristic of papillary carcinoma. There were no three-dimensional, branching papillae with fibrovascular cores. CONCLUSION: Although macrofollicular variant of papillary carcinoma is very rare, fine needle aspiration cytology may contribute to its correct diagnosis. PMID- 9850658 TI - Fine needle aspiration cytology of clear cell sarcoma of the kidney. A case report. AB - BACKGROUND: Fine needle aspiration cytology is an established method for cytodiagnosis. Its application is particularly suited to the pediatric age group. A correct cytodiagnosis of especially large tumors can result in timely cytoreductive chemotherapy, thereby facilitating surgery. While the cytomorphology of Wilms' tumor is well documented, there is a paucity of literature concerning other uncommon pediatric renal tumors. CASE: A 2-year-old female underwent surgery for a renal mass following aspiration cytology. The histologic diagnosis was clear cell sarcoma of the kidney (CCSK). The cytologic smears were cellular and pleomorphic. The main feature of cytologic interest was the presence of deep nuclear indentations and grooves in many of the tumor cells. CONCLUSION: The cytologic features of CCSK are distinct and different from those of other renal tumors in children. Its recognition in cytology is important because its behavior is more aggressive than that of Wilms' tumor. A correct diagnosis can result in the institution of appropriate treatment. PMID- 9850659 TI - Fine needle aspiration of sclerosing lymphocytic lobulitis of the breast. A report of two cases. AB - BACKGROUND: Sclerosing lymphocytic lobulitis (SLL) was described in 1948 by Soler and Khardori as fibrotic disease of the breast with histologic features similar to those of Hashimoto's thyroiditis. Associations of this process with thyroiditis, artropathy and diabetes mellitus have been seen. CASES: We report the fine needle aspiration biopsy (FNAB) findings of two cases of SLL seen in our service. Both patients showed a breast nodule suspicious for malignancy. The cytologic changes vary according to the stage of the disease, from abundant lymphocytes and scanty fibrosis (case 1) to the presence of a few lymphocytes, epithelioid fibroblasts and significant sclerosis (case 2). CONCLUSION: The cytologic changes seen in FNAB are sufficient to indicate the diagnosis of SLL. The differential diagnosis has to be made with primary breast lymphoma, periductal or perilobular inflammation and different types of sclerosing breast lesions, depending on the stage of the disease. PMID- 9850660 TI - FNAC guided by computed tomography in the diagnosis of primary pancreatic adenosquamous carcinoma. A report of three cases. AB - BACKGROUND: Pancreatic adenosquamous carcinoma (ASqC) is an unusual histologic subtype of nonendocrine neoplasia of the pancreas. Although fine needle aspiration cytology (FNAC) is now accepted as a reliable procedure for the diagnosis of pancreatic malignancies, many of these unusual tumors are still diagnosed after surgery or at necropsy. CASES: Between January 1995 and July 1996, 3 of 35 primary pancreatic malignant tumors were diagnosed as ASqC based on computed tomography-guided FNAC. After cytologic diagnosis, all three patients were treated with neoadjuvant chemotherapy and radiotherapy. Two patients completed the treatment and underwent a surgical pancreatic-duodenectomy with antrectomy. The remaining patient is currently under treatment. That patient had a highly infiltrative pancreatic mass that affected the muscular small bowel wall. An endoscopic biopsy was performed. The cytologic diagnosis was confirmed by histology in all cases. Immunohistochemically both components, squamous and glandular, showed reactivity for several keratins, while only the glandular pattern was reactive with carcinoembryonic antigen (CEA). CONCLUSION: FNAC is an accurate, rapid and sensitive tool in the diagnosis of ASqC of the pancreas. We recommend a careful search for both malignant components. Immunoreactivity for CEA can be of help in the detection of the glandular component of this tumor. PMID- 9850661 TI - Adenocarcinoma in situ of the fallopian tube. A case report. AB - BACKGROUND: So far only a few cases of carcinoma in situ of the fallopian tube have been reported, but its detailed clinical and pathologic findings, including cytology, have not been fully described. CASE: A 70-year-old female was admitted to our hospital because of irregular genital bleeding. Endometrial smear revealed a small number of atypical cells with a clear background. Hysterectomy, bilateral salpingo-oophorectomy and omentectomy were performed. Grossly, a grayish white papillary tumor, measuring 1.5 x 1.0 cm, was observed within the lumen of the left fallopian tube. Microscopically, the diagnosis of papillary adenocarcinoma in situ of the left fallopian tube was made according to 1992 International Federation of Gynecologists and Obstetricians fallopian tube staging. CONCLUSION: Although endometrial brush cytology is not sensitive enough to detect a primary carcinoma of the fallopian tube, our case indicates that it may contribute useful information on extrauterine diseases and can detect a stage 0 cancer of the fallopian tube. Clinicians, as well as pathologists, should consider the possibility of fallopian tube cancer if cervical or endometrial cytology shows atypical cells with papillary patterns with a clear background but endometrial curettage cannot prove malignancy. PMID- 9850662 TI - Sertoli cell tumor of the testis. Report of a case with imprint cytology findings. AB - BACKGROUND: Sertoli cell tumor of the testis is a relatively rare neoplasm, and few reports concerning its cytologic findings have been previously documented. CASE: A 41-year-old male had Sertoli cell tumor in the left testis. An imprint smear revealed tumor cells showing a less cohesive or singly dispersed cell pattern. The cytoplasm was faintly stained, and most tumor cells were naked. The nuclei were oval or short and spindle in shape, and some of them showed coffee bean nuclei or nuclear indentation. CONCLUSION: Coffee bean nuclei can be observed in Sertoli cell tumor. PMID- 9850663 TI - Fine needle aspiration of signet-ring cell lymphoma. A case report with differential diagnostic considerations. AB - BACKGROUND: Signet-ring cell lymphoma is an unusual morphologic variant of non Hodgkin's lymphoma that, although well described histologically, is scarcely mentioned in the cytology literature. Its main significance lies in its potential for diagnostic confusion with more common lesions containing signet-ring cells. CASE: A 50-year-old, white male presented with a two-month history of persistent cervical lymphadenopathy and fatigue. Fine needle aspiration of a 2-cm, left, submandibular lymph node revealed classic signet-ring cells among small and large lymphoid cells. Also noted were multivacuolated cells. The background of the smears showed many vacuolated structures analogous to the lymphoglandular bodies seen in lymphoid proliferations without signet-ring cells. CONCLUSION: The differential diagnosis of signet-ring cell lesions by fine needle aspiration includes signet-ring cell lymphoma, sinus histiocytosis and metastatic adenocarcinoma, liposarcoma and melanoma. When confronted with such an aspirate, additional material should be obtained for immunocytochemical or flow cytometric analysis. PMID- 9850664 TI - Cystic lesion of the parotid gland with squamous metaplasia mistaken for squamous cell carcinoma. A case report. AB - BACKGROUND: The diverse range of diseases that affect the salivary glands may lead to problems and pitfalls in cyto-diagnosis. While false negative diagnosis of cystic salivary gland tumors is well known, false positive cytodiagnosis in nonneoplastic salivary cysts is less well documented. CASE: An 85-year-old female presented with a painless left parotid gland swelling of three months' duration. Fine needle aspiration cytology yielded fluid, smears of which showed keratinizing squamous cells with nuclear atypia leading to a cytologic diagnosis of cystic squamous cell carcinoma. A total radical parotidectomy followed. Histopathologic study showed cystic dilatation of many of the salivary ducts, which were lined with metaplastic squamous epithelium that showed atypia. There was no evidence of squamous cell carcinoma. CONCLUSION: Squamous metaplasia is known to occur in benign salivary gland lesions, such as pleomorphic adenoma and Warthin's tumors, as well as in salivary duct cysts and necrotizing sialometaplasia. However, atypical squamous metaplasia of salivary duct cysts mimicking squamous cell carcinoma on cytology is unusual. PMID- 9850665 TI - Ectopic breast tissue on the vulva diagnosed by fine needle aspiration. PMID- 9850666 TI - Management of suboptimal cytologic smears: persistent inflammatory smears. PMID- 9850667 TI - Criteria for cytologic reporting of breast fine needle aspiration. PMID- 9850668 TI - In situ detection of HPV in a cervical smear with in situ hybridization. PMID- 9850669 TI - Fine needle aspiration cytology and histopathology of crystalloids in a cystic lesion of parotid salivary gland. PMID- 9850670 TI - Imprint cytology of lipoma with chondroosseous metaplasia. PMID- 9850671 TI - Pleomorphic lipoma on fine needle aspiration cytology. PMID- 9850672 TI - Acute myelomonocytic leukemia initially presenting as bilateral renal enlargement and diagnosed by FNAC. PMID- 9850673 TI - Diffuse large B-cell lymphoma with sclerosis of the thymus. PMID- 9850674 TI - Mammography screening for breast cancer in Copenhagen April 1991-March 1997. Mammography Screening Evaluation Group. AB - A biennial mammography screening programme started for all women aged 50-69 in the municipality of Copenhagen, Denmark, on 1 April 1991. We report here on the outcome of the first three invitation rounds. Copenhagen has approximately 40,000 female residents aged 50-69. The Copenhagen Municipality IT Service runs the data files and issues the invitations based on the daily updated population register. Screening takes place at a special clinic at Bispebjerg Hospital, and since 1 September 1996 all assessment and surgery take place at Rigs-hospitalet. The first invitation round covered the period April 1991 to April 1993, the second invitation round the period May 1993 to May 1995, and the third invitation round the period June 1995 to March 1997. By now in total 120,772 invitations have been issued, 84,036 screening tests have been performed, 4110 women have been assessed with additional tests, 1057 women have undergone surgery, and 697 cases of invasive breast cancer cases or carcinoma in situ have been detected. The number of women screened during each of the three invitation rounds as a percentage of the target population aged 50-69 was 71%, 65% and 63%, respectively. It is possible for a woman to notify the programme if she does not want to be invited, and this caused the coverage to decline. The participation rate among the invited women was 71%, 69% and 70%, respectively. Ninety percent of those screened the first and second times and invited the third time participated. The number of screen detected invasive breast cancers of carcinoma in situ cases per 1000 participants during each invitation round was 11.9; 6.3; and 6.1, respectively. When compared with the incidence of invasive breast cancer in Copenhagen before screening, the ratios were 4-7 for all first time screened and 2 for all second or third time screened. Among the 697 screen detected cases, 84% were invasive breast cancers and 11% carcinoma in situ in women not previously known with breast cancer, and 5% were second primary breast cancers. Fifty two invasive interval cancers were found when women who tested negative during the first invitation round were followed up for two years in the Danish Cancer Register and the files of invasive breast cancers of the Danish Breast Cancer Cooperative Group. Compared with the incidence of invasive breast cancer in Copenhagen before screening this gave a proportionate interval cancer rate of 0.34 (95% confidence interval 0.26-0.45). Eight percent of the women who ever participated in the programme had experienced at least one false positive screening test, and 10% of the women who participated all three times in the programme had experienced at least one false positive test. The assessment was an efficient procedure for the sorting out of the false positive screening tests. During the third invitation round, three quarters of those who underwent surgery had either invasive breast cancer or carcinoma in situ. The participation rate in the present programme is relatively low as are the participation rates in other large cities. However, the programme is well accepted among those who participate, as a high proportion of these women come back. The detection rates compared with the previous incidence of invasive breast cancer indicate a good performance of the programme, and so does the proportionate interval cancer rate following the first invitation round. The programme is thus functioning well. The relatively low coverage and the flow in the target population limit the reduction in breast cancer mortality to be expected from the screening programme in the Copenhagen municipality. PMID- 9850675 TI - Ancient and recent horizontal transfer events: the origins of mitochondria. PMID- 9850676 TI - Bacteriophage and gene transfer. PMID- 9850677 TI - Broad-host-range plasmids and their role in gene transfer in nature. PMID- 9850678 TI - In situ detection of gene transfer in a model biofilm engaged in degradation of benzyl alcohol. PMID- 9850679 TI - Transposons as gene haulers. PMID- 9850680 TI - The potential of integrons and connected programmed rearrangements for mediating horizontal gene transfer. AB - Site-specific recombination of integrons, mediates transfer of single genes in small genomes and plasmids. Recent data suggest that new genes are recruited to the cassettes--the units moved by integrons. Integrons are resident in a class of transposons with pronounced target selectivity for resolution loci in broad host range plasmids. A resulting network of programmed transfer routes, with potential offshoots reaching into eukaryotic cells, may channel genes to unexpectedly remote organisms. It has previously been observed that the conjugation apparatus of the broad host range plasmid R751 (IncP) which contains transposon Tn5090 harbouring an integron, promotes horizontal genetic transfer between bacteria and yeast. Furthermore, it is well known and fundamental for widely used gene replacement technologies, that site-specific recombination systems (e.g. Cre-lox of bacteriophage P1) related to the integrons are functional in higher eukaryotes. It seems very clear that integrons and associated programmed transfer mechanisms have high significance for the dissemination of antibiotic resistance genes in bacteria whereas further studies are needed to assess their importance for spreading of arbitrary genes in a wider range of host systems. PMID- 9850681 TI - Bacterial gene transfer by natural genetic transformation. PMID- 9850682 TI - Conjugation in gram-positive bacteria and kinetics of plasmid transfer. PMID- 9850683 TI - Competence for natural transformation in Neisseria gonorrhoeae: a model system for studies of horizontal gene transfer. AB - A combined effort integrating studies of gonococcal Tfp biogenesis, the data made available from the gonococcal genome sequence project and applied molecular genetics have been used to identify the fibrillar filaments themselves, the PilT protein and the ComP protein as essential components for the DNA uptake phase of competence for natural transformation. Our ongoing studies are focused on identifying and understanding the complex interactions which exist between these essential constituents. These studies may be relevant not only to the early steps of genetic transformation but also to the two other venues for horizontal gene transfer based on recent findings. First, the thin pili of IncI1 conjugal plasmids required for liquid mating belong to the type IV family of pili (Yoshida et al., 1998). Secondly, type IV pili are required for lysogenic conversion of Vibrio cholerae by a filamentous phage encoding cholera toxin (Waldor and Mekalanos, 1996). How these highly conserved surface organelles contribute to such diverse forms of DNA translocation across membranes remains to be seen. PMID- 9850684 TI - Foreign DNA in mammalian systems. PMID- 9850685 TI - Genome instability. PMID- 9850686 TI - Mobile drug resistance genes among fish bacteria. PMID- 9850687 TI - Barriers to horizontal gene transfer by natural transformation in soil bacteria. AB - Bacteria can utilize horizontally transferred DNA from other bacterial species to adapt and evolve to their changing environments. Natural transformation is a process that allows bacteria, which are able to express a regulated physiological state of competence, to take up and integrate free DNA from their surroundings. This uptake of DNA does not necessarily depend on DNA sequence, thus, indicating the potential of gene transfer from diverged donor organisms. Barriers active against such interspecies transfer are present at different phases of the transformation process. The functionality of these barriers will be discussed, and seen in relation to mechanisms that may enable bacterial cells to respond to environmental stress by adaptive evolution. PMID- 9850689 TI - [Acquired cytomegalovirus infections in immunocompetent adults: 116 cases]. AB - OBJECTIVE: To analyse retrospectively the clinical, biological and epidemiological features of cytomegalovirus (CMV) infection in the immunocompetent host. PATIENTS AND METHODS: A retrospective study was conducted in 116 cases of CMV infection (74 inpatients, 42 out-patients) collected from 1981 to 1997 in a university hospital. Diagnostic was established on serological criteria in all cases. RESULTS: Fever was observed in all cases but one (mean duration: 21 days). The most frequent symptoms were headache (51%) and myalgia (46%). Splenomegaly was the most frequent sign (36%). Pulmonary interstitial opacities on chest x-ray were found in 8.5% of patients. Pulmonary (1 case) and neurological (2 cases) complications occurred. Concomitant HIV primary infection was observed in 2 patients. Mononucleosis and ALAT and LDH elevations were observed in 95%, 85% and 95% of cases respectively. Viremia was positive in 79% (30/38). pp65 antigenemia was useful to establish the diagnosis in 9 patients. Antibiotics were prescribed in 46% before diagnosis was established. CONCLUSION: CMV infection in the immunocompetent host is a frequent and rarely complicated disease. pp65 antigenemia should be evaluated more in this setting. Most hospitalizations due to this affection could be avoided. PMID- 9850690 TI - [Congenital quadricuspid aortic valves]. AB - BACKGROUND: Quadricuspid aortic valve is an uncommon congenital anomaly. We report a case in a patient hospitalized for heart failure. CASE REPORT: A 62-year old patient with exercise-induced dyspnea was hospitalized for heart failure. Heart auscultation revealed a diastolic aortic murmur attributed to a quadricuspid aortic valve evidenced at echocardiography. DISCUSSION: Quadricuspid aortic valves usually have three cusps of equivalent size and a small fourth cusp between the right coronary cusp and the non-coronary cusp. Aortic regurgitation is usually observed requiring valve replacement in 50% of the cases. The anomaly may be associated with other congenital anomalies of the coronary arteries warranting systematic coronarography prior to valve replacement. PMID- 9850691 TI - [Pneumatosis cystoides intestinalis in systemic diseases: 3 cases]. AB - BACKGROUND: Pneumatosis cystoides intestinalis (PCI) is usually a complication of digestive tract or respiratory tract diseases, but rare cases have been described in systemic diseases, mainly systemic sclerosis. CASE REPORTS: Three patients, one with temporal arteritis and two with polyarteritis nodosa (complicating rheumatoid arthritis in one case) were treated by prednisone. All three developed PCI, complicated in one case by a retropneumoperitoneum. Medical treatment led to a favorable outcome in all cases. DISCUSSION: Sixty-two cases of PCI have been reported in patients with various systemic diseases (systemic sclerosis, systemic lupus erythematosus, mixed connective tissue disease, dermatopolymyositis, polyarteritis nodosa, rheumatoid arthritis, Sjogren's syndrome, amyloidosis). Systemic sclerosis is the most frequent condition (45%). In the other cases, corticosteroid therapy or digestive tract vasculitis are the main causal factors. Outcome is usually favorable with medical treatment. Laparotomy is rarely needed. PMID- 9850692 TI - [Sudden awakening of a parasite...]. PMID- 9850693 TI - [Association of celiac disease and antiphospholipid syndrome]. PMID- 9850694 TI - [MALT-type primary pulmonary lymphoma. Recurrence 17 years after surgical excision]. PMID- 9850695 TI - [Carrington's disease associated with naso-sinusal polyposis]. PMID- 9850696 TI - [Interdigital periarterial sympathectomy: treatment of severe and refractory Raynaud's phenomenon]. PMID- 9850697 TI - [Are there still indications for tubal surgery in infertility?]. AB - In vitro fertilization (IVF) has revolutionalized management strategies proposed for infertility to the point where some have suggested that tubal surgery is no longer indicated. The objective being to obtain the largest number of viable births at minimum cost and maximum patient comfort, we strongly believe that tubal surgery and IVF are complementary tools for patients with tubal infertility. The choice depends on the severity and the localization of the lesions. For example, in patients with hydrosalpinx (the most frequent tubal lesion) and healthy or moderately altered mucosa, the rate of in utero pregnancies after tubal plasty by laparoscopic salpingostomy (53% in our series) is quite comparable with the rate after IVF. In this large group, tubal surgery should be proposed first. Tubal surgery may also be indicated in other less frequent lesions such as proximal obstructions where we have obtained excellent results (56% live births within 2 years of surgery). Of course, the final decision is made by the couple, well informed of the advantages and disadvantages of each technique. Performed by a well-trained team with extensive experience in reproduction, tubal surgery, particularly with the development of micro instruments for laparoscopy, remains a treatment of choice for patients with tubal infertility. PMID- 9850698 TI - [Leukotriene receptor antagonist effective in chronic asthma]. PMID- 9850699 TI - [Forum on bacterial resistance]. AB - Recently, in daily newspapers and on television, attention of the audience has been focused on the overuse of antibiotics and on the role it plays in the emergence and dissemination of resistance mechanisms in the human environment. The role of food from animal origin in relation to the use of antibiotic resistance, infectious diseases, medical practice and ENT infections have accepted to answer a series of questions concerning risks versus usefulness of antibiotic usage. From the answers, we may note convergent views and discrepancies: (i) there was agreement concerning the unnecessary prescription of antibiotics in rhinopharyngitis and few other common viral infections; (ii) the risk of misuse of antibiotics in patients with poor compliance and further risk of erroneous self prescription of the remaining tablets has been cited; (iii) in the problem of resistance resulting from growth promoting antibiotics in animals, it has been experimentally shown that from 2 bacteria of the same species introduced in the animal gut, one susceptible, the other resistant, the latter will be eliminated by means of the "barrier effect"; similarly in case of transfer of resistance from an exogenous bacteria to a "resident" organism of the gut, the latter will be eliminated by the homologous susceptible ones; only an antibiotic therapy may confer importance to the resistant bacteria. In this respect, care should be taken for resistance spread such as that concerning penicillin-resistant pneumococci and surveillance and control of resistance mechanisms has become necessary. However we should look with reluctance at the diffusion of inevitably simplified and truncated information from Media, showing the negative aspects of antibiotics only. Moreover, as underlined by the expert from the Institut Pasteur, there are new perspectives in the development of effective new agents based on the modern "genomic" research. PMID- 9850700 TI - [Nephrotoxicity of ritonavir]. AB - A RECOGNIZED COMPLICATION: Ritonavir is an antiprotease used in the treatment of HIV-positive patients. Among the known side effects, nephrotoxicity can be severe. We have observed acute renal failure in 8 patients. CIRCUMSTANCES: Renal failure occurs early after introducing ritonavir (3-21 days). It is often severe with major creatinine elevation. One patient was dialyzed for 16 days. In these patients, saquinavir was usually associated with ritonavir. RITONAVIR ALONE: We retrospectively analyzed creatinine levels in 87 patients treated with ritonavir without saquinavir. Twelve of these 87 patients (13.7%) developed renal failure. Creatinine clearance (Cockcroft) was reduced 116 to 71 ml/min in 12 patients. Finally, it was demonstrated in 6 patients that ritonavir can reduce creatinine clearance by 25% after only 3 days of treatment. VIGILANCE: Ritonavir has a known nephrotoxic potential. Acute renal failure may be severe and can occur with ritonavir alone or in combination with saquinavir. The pathogenic mechanism has not been demonstrated from renal biopsies or experimental studies. Renal function should be followed in these patients and risk factors controlled. PMID- 9850701 TI - [Pneumatosis cystoides intestinalis]. AB - AIR-FILLED CYSTS: Pneumatosis cystoides intestinalis (PCI) is a benign air-filled cystic formation lying in submucosal or subserous digestive tissue. PCI has been reported all along the digestive tract. CLINICAL SIGNS: Manifestations of PCI vary greatly. Some cases are asymptomatic, while others are revealed by abdominal pain or pneumoperitoneum. Outcome is usually favorable. EXPLORATION: The plain radiogram of the abdomen shows gaseous cysts of various forms lying between the liver and the diaphragm. Pneumoperitoneum may be present. Computed tomography is the ideal diagnostic test. Endoscopy may be useful for colonic localizations. PATHOGENESIS: Many causes have been suggested and debated. Fifteen percent of all cases of PCI are idiopathic. In the other cases, digestive tract or respiratory tract diseases, are usually the underlying cause. Exceptionally systemic disease may be associated with PCI, particularly systemic sclerosis. TREATMENT: Surgery should be reserved for particularly severe cases. PMID- 9850703 TI - [Observing in HIV infections]. PMID- 9850702 TI - [Human coronavirus infections: importance and diagnosis]. AB - POORLY-KNOWN VIRUS: Coronaviruses, so named because of their sun-ray-like aspect, were discovered in the sixties. The biology of these RNA viruses is complex and poorly understood. KNOWN PATHOGENS: Coronaviruses are known pathogens in veterinary medicine, causing disease states in several domestic species. In human medicine, they can cause benign respiratory infections, but few laboratories include coronaviruses in their routine diagnostic tests. SUSPECTED PATHOGENS: There is some data in the literature suggesting coronaviruses might be implicated in more severe diseases including multiple sclerosis, necrotizing enterocolitis, and lower respiratory tract infections, particularly in infants. IMPROVING DIAGNOSTIC METHODS: Due to the lack of reliable and sensitive diagnostic techniques, it is impossible to date to correctly assess the medical impact of these ubiquitous and endemic viruses. Molecular biology techniques enabling detection of human coronavirus infections should be applied to verifying the suspected implication of these viruses in diverse disease states. PMID- 9850705 TI - [Magnetic resonance of the breast]. PMID- 9850704 TI - [From science's archives, a centennial: Rutherford and the discovery of alpha and beta rays]. PMID- 9850706 TI - Diagnostic imaging of bullous pulmonary disease. A review. PMID- 9850707 TI - [Tumors and other lesions composed of adipose tissue. Integrated diagnostic imaging]. PMID- 9850708 TI - [Complex maxillofacial trauma: diagnostic contribution of multiplanar and tridimensional spiral CT imaging]. AB - INTRODUCTION: Adequate radiologic assessment of the maxillofacial trauma patient is the basis for planning reparative surgery. We investigated the yield of the integration of axial CT with multiplanar (MP) and three-dimensional (3D) reconstructions with the Spiral technique. MATERIAL AND METHODS: Thirty-five patients (21 men and 14 women, mean age: 31.2 years) with complex maxillofacial traumas were submitted to Spiral CT. Images were acquired with 2-3-mm collimation, 1:1 to 2:1 pitch, 210 mAs, 120 kV, 15-24 s Spiral scan, RI = 1. 3D reconstructions were always obtained and used to guide MPRs targeted on the single injury. The examinations were retrospectively given a score, namely 1 if 3D and MPR yielded no more important diagnostic information than axial CT, 2 if 3D and MPR permitted better detailing of some axial CT findings and thus improved image reading, and 3 if 3D and MPR showed new injuries missed on axial images. Then, a radiologist and a maxillofacial surgeon reviewed the 3D images together only to assess in which cases they were useful to optimize surgical planning. RESULTS: Nine cases (25.7%) scored 1, seventeen (48.6%) scored 2 and nine (25.7%) scored 3. MP and 3D reconstructions were useful or determinant (2 + 3) in over 74% of cases. 3D images made surgical planning easier in 15 of 32 surgical patients (46.8%), allowing the surgeon a better panoramic view of the complex fracture. CONCLUSIONS: The greatest advantage of multiplanar imaging is the improved depiction of skeletal injuries along a horizontal plane, paralleling that of axial scans. The depiction of fractures of cribrum and of orbital roof and floor was particularly useful from a clinical viewpoint. MPRs clearly depicted herniation and incarceration of the lower rectus muscle in blow-out fractures. MP and 3D reconstructions better defined the presence and grade of displaced bone fragments in nearly vertical structures, such as the upward branches and coronoid apophysis of the mandible. 3D images alone never showed any more fractures than those seen on axial and MP images. 3D images are especially useful to the surgeon because they permit the panoramic depiction of the fracture complex, which facilitates treatment planning. The radiologist can use 3D reconstructions to guide the acquisition of targeted MP reconstructions, for better diagnostic yield. The Spiral technique, with its pitch increases up to 2, permits to limit the radiation dose while preserving the quality of postprocessing reconstructions. Finally, decreasing the execution time is important in multiple trauma patients who are often clinically unstable or have damaged vital organs. PMID- 9850709 TI - [Magnetic resonance imaging of seronegative sacroiliitis]. AB - INTRODUCTION: The inflammatory involvement of the sacroiliac joint is frequent during seronegative spondylarthritis. The clinical diagnosis of sacroiliitis may be very difficult, especially in the early stage, because joint motion cannot be assessed directly and the clinical picture is very similar to that of lumbar pain. Conventional radiography is negative as long as the structural change in the joint is limited to the synovial membrane and the cartilage (early stage). Computed Tomography (CT) also has many drawbacks, and thus the changes can be shown only when chondritis and enthesitis have already damaged the bone. The disease onset is usually preceded by a long latency; early diagnosis is needed for a proper and timely treatment, which can be made only with a highly sensitive and specific technique. We investigated the diagnostic accuracy of MRI in the early detection of sacroiliitis during seronegative spondylarthritis. MATERIAL AND METHODS: Forty patients with suspected sacroiliitis and negative radiographic findings were submitted to MRI; thirty-seven of them were HLA B27 positive. MRI was performed with a .5 T superconducting unit; T1-weighted SE, T2-weighted FSE, T2* GE, and STIR images were acquired on the oblique coronal plane parallel to the anterior sacrum. Ten asymptomatic volunteers were also examined as a control group. RESULTS: An irregular sacral border and marrow changes at the insertion of the sacroiliac ligaments were seen in 3/10 asymptomatic volunteers. MRI was negative in 7/40 patients, while the synovial compartment was replaced by some tissue with low signal intensity of T1 and high signal on T2 in the other 33 patients; this finding was referred to synovial pannus. Persisting low-signal foci were seen in the synovial compartment in 16/33 patients, which were referred to spared cartilage. High-signal regions were depicted at the bone periphery in 9/33 patients, which areas were consistent with bone erosion; the subchondral bone was markedly hypointense in 5 of these patients, indicating sclerosis. Finally, diffuse high signal intensity was found in the bone marrow in 3/33 patients and referred to infectious sacroiliitis. CONCLUSIONS: MRI appears the method of choice for the early detection of seronegative sacroiliitis because it can show the early changes in cartilage and subchondral bone, filling the gap between the onset of symptoms and radiographic evidence. Moreover, MRI uses no ionizing radiations and makes therefore a precious tool for the diagnosis and follow-up of young patients, hopefully decreasing the use of CT which however provides better detailing of bone and bone degeneration. PMID- 9850710 TI - [Significance of computerized tomography in the diagnosis of post-traumatic proximal carpal instability]. AB - INTRODUCTION: Proximal carpal instability is a painful condition characterized by early or late loss of radioulnar joint (RUJ) congruence not affecting the normal bone alignment of the two carpal rows. The joint incongruence or (incomplete) dislocation which leads to proximal instability is caused by many traumatic and nontraumatic events. The diagnosis of (incomplete) dislocation of the distal RUJ may be extremely difficult to make at conventional radiography because such injuries can be seen only when the lateral joint projection is perfect; otherwise the diagnosis is not reliable. CT is the only imaging tool diagnosing the grade of distal RUJ congruence independent of the examination technique. MATERIAL AND METHODS: We studied the radiocarpal complex conditions leading to proximal instability at the Rizzoli Orthopedic Institute from December, 1995, through December, 1997. In all, 389 cases were seen, 376 from trauma and 13 of nontraumatic origin. Each injury was studied with conventional radiography, CT, and MRI. Radiography was performed in two projections, namely the posteroanterior one with hand extension and the lateral one with the forearm in neutral position and the elbow bent at 90 degrees; the projections were repeated whenever a cast brace was applied. Unenhanced CT was performed for comparison with the patient prone and the forearm and wrist in prone and neutral position, as well as with the patient, forearm and wrist supine. Three criteria of electronic image processing were adopted for the RUJ studies in the 3 projections: radioulnar lines, congruence, and epicenter. MRI was always performed after conventional radiography and CT. Only the involved radiocarpal region was studied; coronal, axial and sagittal images were acquired with T2-weighted GE and T1-weighted SE sequences. RESULTS: Proximal instability was found in 17 of 389 patients; it was early in 13 and late in 4 of them. The comparison of radiographic and CT results showed that the former method is unreliable, with 53% false negatives. Pain, a cast brace, congenital or acquired deformities of distal radius and ulna and patient mispositioning by the radiology technician can change the rotation of the forearm, wrist and hand and make a perfect laterolateral projection in neutral position unfeasible, which affects the radiographic diagnosis. Conversely, CT showed its extreme efficacy in assessing the distal RUJ congruence with no false negatives independent of the RUJ rotation and of instability type and grade. CONCLUSIONS: Conventional radiography is a poorly reliable tool for the diagnosis of joint incongruence and its grade. In contrast, CT can diagnose a RUJ (incomplete) dislocation easily and unquestionably, thanks to its axial capabilities, even when adequate radiographic studies would be unfeasible. If the anteroposterior projection of the radiocarpal complex shows a congenital or acquired deformity of distal radius, the lateral projection can be skipped and a CT scan in prone, neutral and supine position performed. The 3 CT criteria quantify incongruence type and grade, and also demonstrate the position of maximum incongruence and its decrease by position. The comparative study of the radiocarpal region makes CT a very useful and valuable tool in congenital instability because its allows the assessment of contralateral radioulnar congruence too. MRI is very useful in the diagnosis of injury or degeneration of the fibrocartilage complex, namely in patients with no bone changes at conventional radiography. PMID- 9850711 TI - [Anterior cruciate ligament tears in athletes: assessment and clinico-diagnostic control with a dedicated system]. AB - INTRODUCTION: We investigated the MR signs of anterior cruciate ligament (ACL) tears and tried to assess the patterns of both the acute and the chronic phases since more MR examinations are currently performed of the knee joint in the hyper and acute post-traumatic stage. MATERIAL AND METHODS: December, 1994, through September, 1997, a hundred and 89 sportsmen with a history of hyper and acute knee trauma were submitted to MRI: one hundred and four of them were followed-up for 3-6 months. The MR examinations were performed with a dedicated unit (Artoscan, Esaote Biomedica, Genoa) using T1-weighted SE, T2-weighted GE and Turbo ME sequences on the sagittal, coronal and axial planes, respectively (3-5 mm slice thickness). RESULTS: Of 189 patients examined in the hyper and acute post-traumatic stage, 101 had a complete ACL tear, 23 had a partial tear and 65 had an enlarged and inhomogeneous ACL: Eighty-five patients underwent surgery, while the other 104 had 3-6 months' follow-up MRI. Twenty-four of the latter had a complete ACL tear, 20 had a partial tear and 60 had no MR finding of ACL interruption, with a more or less inhomogeneous ligament. DISCUSSION: ACL tears are difficult to demonstrate in the acute phase because of perilesional swelling, synovitis and hemorrhage; high-contrast sequences on the axial plane are therefore required. In the chronic stage, an uninterrupted ACL does not necessarily imply that function is preserved, as well as the MR finding of a deflected ligament does not necessarily imply altered biomechanics. CONCLUSIONS: MRI of the knee joint is increasingly used to study hyper and acute ACL tears thanks to the availability of dedicated units. The MR study of ACL tears is not meant to refer the patients to surgery, which pertains to the physical examination, but it allows to assess lesion severity and to show associated injuries in other knee structures. PMID- 9850712 TI - [Botallo's duct calcification in children: radiologic findings]. AB - INTRODUCTION: Calcification of the ductus and ligamentum Botalli with no cardio respiratory symptoms is normally a sign of their closure. However, as the calcification can be associated with a patent duct or a ductal aneurysm, its presence must not be misinterpreted. The frequency and the different patterns of this calcification must be known, as well as is diagnostic relevance. It is also important to differentiate it from other types of upper mediastinal calcification. MATERIAL AND METHODS: Thirty-two cases of ducts Botalli calcification were collected over 16 years, during which time period 38,476 children (24,095 males, 14,381 females) were submitted to chest radiography. Patient populations were divided into the following age-groups: preterms newborns; 2-3 months; 4-6 months; 7-12 months; 2-3 years; 4-6 years; 7-10 years; 11 years on. We studied the ductus Botalli according to the following parameters; conspicuity, location, size and shape of the calcification; prevalence of the radiographic finding in 1,000 patients by gender and age; persistence of the finding in follow-up examinations. RESULTS: The calcification was found in left mediastinal site at the posterior aspect of the 4-6th ribs, ranging 1-6 mm in length, and 1-4 mm in width. It had a round, comma-like or elongated shape. In all, 32 cases of ductal calcification (.83/1000) were found: 14 (44%) were in males (age range 7 months-12 years), 18 (56%) in females (age range 11 months-10 years). Age range analysis shows that the calcification is distributed more evenly in the male population with the highest number of calcified ducts (5 = 94/1000) found in the 4-6 year age group and a relative prevalence in the age group 7-12 months (2 cases = 1.05/1000). A marked prevalence is found in females 4-6 years old (11 cases = 3.8/1000). The duct calcification was an occasional finding during chest radiography for other, more severe conditions in 9 of 14 males and in 6 of 18 females. CONCLUSIONS: No data can be found in the literature on the frequency of ductus Botalli calcification at chest radiography: our results indicate it to be less than 1/1000 (.83/1000). We have found a prevalence of ductus calcification (1.25/1000) in females even though females are less numerous in our population. The ductal calcification is often associated with severe, mostly hematologic, diseases. We believe that the conspicuity of the calcification is due to its density more than to its shape and size. Literature data indicate that ductus calcification may still be visible 6 months to 8 years after its discovery: our results, although on a small series, confirm this observation. The clinical significance of the calcification is still unclear. PMID- 9850713 TI - [Computerized tomography findings in intestinal carcinoid]. AB - INTRODUCTION: We studied the CT patterns of abdominal carcinoid tumors. MATERIAL AND METHODS: Eight patients with carcinoid tumors were examined with CT. The tumors were in the ileum (2 patients), duodenum (1), colon (1), and mesentery (4). Pathologic confirmation was obtained at surgery in all patients. The symptoms were the carcinoid syndrome in 2 patients, abdominal pain in 4, jaundice in 1 patient and a right lower quadrant mass in 1. All patients were examined with(out) i.v. contrast agent administration; 6 patients received oral contrast material and 2 were submitted to water enema, to enhance visualization of the distal ileum and colon. RESULTS: CT identified the tumor in all patients but correctly defined its site in 7 cases only. CT showed a small mass in the distal common bile duct in 1 case, which postoperative histology diagnosed as a malignant carcinoid tumor of the duodenum. The CT findings of carcinoid tumors were a rounded mesenteric mass displacing bowel loops in 4 cases, an infiltrating colonic tumor in 1 case, an apparently intracholedochal mass in 1, a large necrotic mesenteric mass in 1 case. CT showed lymph node involvement in 2 cases, but no liver metastases were found. A carcinoid tumor was diagnosed in 4 patients based on the typical CT finding of a mesenteric mass with radiating soft-tissue density bands resulting in a stellate pattern. No preoperative diagnosis was possible in some other cases because the CT patterns were aspecific and mimicked those of other lesions, namely of tumor of the distal common bile duct, adenocarcinoma of the right colon, submucosal tumor of the distal ileum, large necrotic mesenteric mass. CONCLUSIONS: CT is a useful tool in the detection of carcinoid tumors. The correct diagnosis can be made based on the characteristic CT finding of a rounded mesenteric mass. In our experience, however, carcinoid tumors may present with other CT patterns mimicking those of other tumors. PMID- 9850714 TI - [Ultrasonography in the diagnosis of enterocutaneous fistula in Crohn's disease]. AB - INTRODUCTION: Perforation seems to be a specific pathologic aspect in some types of Crohn's disease. Fistulae are caused by a transmural extension of a fissure and/or an ulcer; they are often multiple and can be internal or external. External fistulae usually occur after surgery and along the scar incision. They are frequently complicated by associated intra-abdominal abscesses. We investigated the accuracy of fistulography by ultrasonography compared to fistulography by X-rays in the diagnosis of enterocutaneous fistulae. MATERIAL AND METHODS: Eight patients resected for Crohn's ileitis and with enterocutaneous fistulae were examined by fistulography with ultrasonography and then by fistulography with X-rays. Disease recurrence was established with ultrasonography on ileum anastomosis in all patients. A7.5 MHz linear transducer was connected to a videotape. Physiologic solution was injected into the cutaneous orifice using a thin rigid catheter to evaluate the communication with the intestinal tract by fistulography. At the same time all patients underwent fistulography with contrast agent injection through a radiopaque catheter. RESULTS: On the same day two radiologists performed the ultrasonographic and the radiologic examinations and had 100% agreement between the two methods. In five of eight patients (62%) there was no intestinal communication. Ultrasonography showed hypoechoic lines turning deeply from the cutaneous surface with no communication with the intestinal tract. Radiology confirmed the ultrasonography diagnosis. In two of five patients the fistulae ended in small abscesses. The patients had been treated with immunosuppressive or parenteral therapy; four of them improved and one underwent abscess drainage. In three of eight patients cutaneous fistulae communicated with the intestinal anastomosis. In one patient there was an abscess with multiple fistulae, one of which communicated with the third duodenal segment. DISCUSSION: The origin, anatomic course and sites of communication of fistulae should be evaluated with conventional barium studies first. These studies may be limited by the fact that the origin of the fistulae could be edematous and prevent contrast opacification, in which cases conventional fistulography or ultrasonographic fistulography should be performed. CONCLUSIONS: Our experience suggests that ultrasonography is a reliable method for detecting intestinal alterations and especially the complications typical of Crohn's disease such as enterocutaneous fistulae. PMID- 9850715 TI - [Computerized tomography, magnetic resonance, and nuclear medicine in the non invasive diagnosis of focal nodular hyperplasia of the liver]. AB - INTRODUCTION: Focal nodular hyperplasia (FNH) is a benign liver lesion requiring a prompt diagnosis and a conservative management. Aim of our study was to prospectively integrate enhanced CT, MRI, nuclear medicine in the noninvasive diagnosis of FNH. MATERIAL AND METHODS: 20 FNH lesions (diameter ranging 1.5-13 cm) in 18 asymptomatic patients were investigated with MRI and nuclear medicine. MRI examinations were performed with a 1.5 T superconducting system (Philiphs NT) by acquiring T1-weighted, T2-weighted, T2-weighted fat-suppressed Turbo Spin-Echo and dynamic MRI sequences, with breath-hold T1-weighted Turbo Field Echo and Gd DTPA. Nuclear medicine included in all cases 99mTc sulfur colloid or 99mTc iminodiacetic acid studies. Diagnostic sensitivity of MRI was compared with that of nuclear medicine, and the sensitivity of the two modalities combined. The definitive diagnosis was made by percutaneous core-needle biopsy (12 lesions), surgery (2 lesions) and longterm follow-up (6 lesions). RESULTS: In 13 lesions larger than 3.5 cm typical findings, such as the central scar and homogeneous pattern, were observed in 9 cases (75%) with enhanced CT, in 10 cases (83%) with unenhanced MRI and in 11 cases (91%) after gadolinium injection. Hepatobiliary scintigraphy showed increased tracer uptake in delayed scans in 10/12 cases (83%) while sulfur colloid studies were diagnostic in 2/12 cases only (16%), showing the same tracer uptake than the surrounding liver parenchyma. In 7 lesions smaller than 3.5 cm, only 1 case showed typical findings; dynamic MRI showed typical early hypervascularity in 5 of the 6 remaining lesions (71%) which persisted on late images. In lesions smaller than 3.5 cm, sulfur colloid studies were diagnostic in 1/7 lesions (14%) and hepatobiliary scintigraphy in 3/7 lesions (42%). Considering all 20 lesions, MRI-nuclear medicine integration performed better than MRI alone. CONCLUSIONS: Enhanced MRI and biliary scintigraphy are important tools to make the final diagnosis of FNH. In small lesions both examinations should performed, but in large lesions MRI may be sufficient to make an unquestionable diagnosis, limiting hepatobiliary scintigraphy to questionable cases. PMID- 9850716 TI - [Dynamic pancreatography with magnetic resonance after functional stimulus with secretin in chronic pancreatitis]. AB - AIM OF THE STUDY: Magnetic Resonance pancreatography (MRP) was performed before and after the intravenous injection of secretin to assess the improvement in pancreatic duct visualization and to perform a dynamic study of the pancreatic exocrine function. MATERIAL AND METHODS: 20 MRP examinations were performed in 18 patients with suspected or known chronic pancreatitis. Coronal T2-weighted half Fourier SSFSE images were obtained with a phased array surface coil. Images were obtained before and up to 10 minutes after the injection of 1 cu/kg b.w. secretin. Quantitative image analysis included main pancreatic duct enlargement over time after secretin injection and the amount of duodenal filling. Qualitative image analysis included: overall image quality improvement, number of pancreatic duct segments visualized, secondary ducts dilation, intraductal filling defects, the presence of pancreas divisum. RESULTS: After secretin injection the overall image quality was judged sufficient in 2 patients and satisfactory in 18 patients. The number of pancreatic duct segments visualized increased from 40/57 (79%) to 57/57 (100%); secondary ducts were visualized in 4 patients before secretin compared to 18 after secretin. The number of stenosis visualized increased from 6 to 9, while intraluminal filling defects increased from 2 to 6. Pancreas divisum was detected in 2 patients after secretin versus 0 before secretin. The main pancreatic duct enlargement was statistically significant in the head of the pancreas (p < .05). Duodenal filling was normal in 13 patients and decreased in 7. DISCUSSION AND CONCLUSIONS: Secretin injection extends the capabilities of MRP in visualizing the morphologic features of pancreatic ducts. The depiction of pancreatic ducts, stenosis, filling defects and pancreas divisum was improved after secretin injection. The exocrine function of the pancreas can be evaluated analyzing the entity and the timing of the duodenal filling. PMID- 9850717 TI - [Computerized tomography and magnetic resonance in the study of retroperitoneal sarcomas]. AB - PURPOSE: We investigated the yield of CT and MRI in the diagnosis and staging of 15 patients with retroperitoneal sarcomas; these lesions are often asymptomatic and need radical surgery to avoid local recurrences. MATERIAL AND METHODS: April 1993, to September 1997, fifteen patients with retroperitoneal sarcomas were examined and then operated on. CT and CT-guided FNAB were always performed, while MRI was performed in 8 patients only. Because of the high risk of sampling errors, the bioptic specimens were immediately evaluated by a pathologist: a second sampling was required in 10 cases and a third sampling in 6 cases. MRI was performed with a .5 T system (GE Vectra); T1- and T2-weighted transverse images of the abdomen were acquired in all patients and additional coronal and sagittal images were acquired for each abnormal region. All patients underwent surgery and the pathologic diagnosis was compared with CT and MR findings. RESULTS: CT always allowed accurate location of the lesions and identification of their components, especially fat deposits. The cytologic examination of FNAB samples allowed the diagnosis of sarcoma in 12 of 15 cases. CT results were compared with MR findings in 8 patients and the latter method had better spatial definition of the abdominal masses, particularly of vascular structures, which is important for surgical planning. DISCUSSION: The present-day imaging techniques are very important to plan the surgical treatment of retroperitoneal sarcomas. In particular, CT has the advantage of high spatial resolution and excellent tissue contrast from abundant retroperitoneal fat tissue; it also permits the cytologic sampling of viable tumor tissue. However the bulk of such lesions often prevents CT from determining the tumor origin, in which case MRI provides better spatial resolution and vascular detailing, which helps surgical planning. CONCLUSIONS: Both CT and MRI are major tools in the diagnosis and staging of retroperitoneal sarcomas because they can accurately define the borders of large tumors and their relationships with surrounding organs. MRI has the advantage of characterizing the blood supply to the mass, but CT is better to guide FNAB. PMID- 9850718 TI - [Ultrasonography-guided percutaneous ethanol injection in large an/or multiple liver metastasis]. AB - INTRODUCTION: Percutaneous ethanol injection (PEI) under sonographic guidance is an effective therapy for hepatocellular carcinoma on cirrhosis, while less favorable results have been reported for liver metastases. Surgery and/or other new treatments (i.e., interstitial thermotherapy) are indicated only for small metastases (< 3 cm) and surgeons no longer perform the palliative debulking of neoplastic masses. PATIENTS AND METHODS: From March, 1994, to December, 1997, thirty-three patients with 62 large (> 3.5 cm) and/or multiple liver metastases, who were not eligible for surgery nor thermotherapy, were treated with one-shot PEI under general anesthesia. The diameter of the nodules ranged 35-92 mm (mean: 39); the lesions were single in 15 patients and localized in both the right and the left lobe in 19 patients. 25-110 ml ethanol were injected per session. Post treatment results were assessed with dynamic or dual-phase spiral CT; therapeutic success was defined as the absence of hyperdense lesion areas. RESULTS: Complete necrosis of the metastases was shown in 10 patients (30.3%). Necrosis rate ranged 70-90% in 21 patients (64%) and was 50% in 2 patients (5.7%). Survival rates were 94%, 80%, 80% and 44% at 12, 24, 36 and 44 months, respectively. No major complications were observed. Seeding of neoplastic cells along the needle tract has been never observed to date. DISCUSSION: Metastasis diameter and number impact on long-term survival. PEI under general anesthesia allows to treat also the patients who are not eligible for other treatments and to inject large amounts of ethanol per session in different tumor areas because metastases usually set on in an otherwise healthy liver. CONCLUSIONS: One-shot PEI can cause major, even complete, tumor necrosis in large and multiple liver metastases. The absence of any important complications and the survival rates in our series seem to indicate that one-shot PEI is effective for tumor debulking in patients not eligible for surgery and other alternative treatments. PMID- 9850719 TI - [Restaging brain computerized tomography after treatment of non-operable lung neoplasms]. AB - PURPOSE: To assess the role of CT brain scans as a routine restaging procedure after primary, aggressive, drug or radiation therapy of unresectable lung cancer. If early, asymptomatic brain metastases are detected and treated, survival could be improved relative to the patients showing brain involvement in a later CT scan performed during the follow-up, at the onset of neurological symptoms. MATERIAL AND METHODS: One hundred patients affected with lung cancer, unresectable on account of histology (small-cell carcinoma) or advanced stage (III, IV) were submitted to chemo- and/or radiotherapy, after a clinical staging including brain CT, which was negative in all patients. Brain CT was also repeated at the end of therapy (restaging), in the absence of any neurological symptom. Further scans were obtained during the subsequent follow-up only when clinical symptoms occurred, suggesting metastases to the brain. Survival values were analyzed in the patients whose brain involvement was detected during restaging, vs those showing symptomatic brain metastases during the follow-up. RESULTS: Only 4 patients had asymptomatic metastases, diagnosed with the restaging brain CT scan. Their survival rate was significantly lower than that of the 20 patients whose brain involvement was shown by a follow-up CT scan, performed after the onset of neurological symptoms. However, death was rarely a consequence of brain metastases: primary or other metastatic sites were involved in the terminal events, in the greatest majority of these cases. DISCUSSION AND CONCLUSIONS: The sudden, asymptomatic brain involvement, detected at restaging CT scan after primary therapy for unresectable lung cancer, does not correlate with a better prognosis than symptomatic metastases, diagnosed later with a follow-up CT obtained performed for clinical suspicion. Therefore the use of restaging CT scan is not warranted, as a routine procedure, except for the clinical trials intended to define optimal treatment schedules. PMID- 9850720 TI - [Radiotherapy of the tonsillar region. Analysis of prognostic factors]. AB - INTRODUCTION: We report our personal experience with the treatment of tonsillar cancers at the Otorhinolaryngology-Radiotherapy Department of Umberto I Hospital, Mestre, Italy. The results were analyzed by tumor site and stage, lymph node involvement, treatment type and patient age. MATERIAL AND METHODS: January, 1987, through December, 1995, we treated a hundred and 25 patients with carcinoma of the tonsil and tonsillar region. Most patients were men (M:F = 4:1), with a mean age of 61.9 years (range: 38-87). The lesions were staged at physical examination, chest radiography, bone scintigraphy. US of the liver and neck, CT and/or MRI of the tonsillar region and neck. Eleven patients were in stage I (8.8%), 26 in stage II (20.8%), 31 in stage III (24.8%) and 57 in stage IV (45.6%). Forty-one patients were submitted to tonsillectomy and more/less massive neck dissection: surgery was not radical-in 14 of them. All patients received gamma-photon radiotherapy with a cobalt unit: the minimum dose was 50 Gy after radical surgery and 60 Gy for exclusive irradiation and after nonradical surgery. The hemiblock field technique was always used with the conventional fractionation (2 Gy/day. 1 fraction/day, 5 fractions/week); the treatment was planned with the Theraplan V05-B method on CT scans. When the tolerance dose was reached, the spinal cord was shielded and the dose compensated with 9 MeV electrons. The treatment was discontinued only when needed, and never for more than 7-10 days. RESULTS: The overall 5-year survival and the 5-year disease-free survival rates were 28% and 45%, respectively; the overall 5-year cause-specific survival rate was 39%. Disease-free survival was 81% in stage I, 52.7% in stage II, 44.2% in stage III and 35.8% in stage IV (p = .005). The 5-year disease-free survival for the patients receiving surgery and irradiation was 62.1%, versus 38.3% for irradiation alone; the rate was 37.6% when neck nodes were involved (N+). One hundred and two patients achieved complete remission (CR), while the other 23 had partial remission (PR). Twenty-eight CR patients recurred; the most common cause of death was failure in primary tumor local control. There were no complications during or after treatment. Secondary lesions were found in 13 patients (10.1%). CONCLUSIONS: Irradiation alone yields fairly good results in early tonsil carcinoma, while the surgery-irradiation combination should be preferred in large tumors. Better results are expected from kinetic and conformal irradiation techniques with 3D calculations on CT and MR images, which should permit to deliver high doses to strictly targeted areas and to reduce side-effects. Other improvements are expected from new combination therapies. PMID- 9850721 TI - [Isolated pancreatic rupture caused by abdominal blunt trauma in a child. Study with spiral computerized tomography]. PMID- 9850722 TI - [A case of disseminated hepato-splenic involvement in infectious mononucleosis. Computerized tomography and magnetic resonance findings]. PMID- 9850723 TI - [Magnetic resonance findings in a case of multiple adenomatous hyperplastic lesions in hemosiderin-containing liver]. PMID- 9850724 TI - [Complete remission of hepatocarcinoma metastasis during palliative treatment with tamoxifen]. PMID- 9850725 TI - [Computerized tomography and magnetic resonance findings in a case of papillary carcinoma of the pancreas]. PMID- 9850726 TI - Cervical ectopic pregnancy treated with uterine arteries embolization and evacuation. A case report. PMID- 9850727 TI - [Carcinomatous degeneration in mullerian duct cyst. Findings with computerized tomography]. PMID- 9850728 TI - [Acute onset of Wilms tumor after minor injury. Assessment with computerized tomography and magnetic resonance]. PMID- 9850729 TI - [Radioprotection]. PMID- 9850730 TI - Role of dystrophin isoforms and associated proteins in muscular dystrophy (review). AB - The membrane cytoskeletal component dystrophin and its associated glycoproteins play a central role in the molecular pathogenesis of several muscular dystrophies, i.e. Duchenne/Becker muscular dystrophy, congenital muscular dystrophy and various forms of limb-girdle muscular dystrophy. Although the most frequent of these disorders, Duchenne muscular dystrophy, is mainly recognized as a disease of skeletal muscle fibers, pathophysiological changes also involve the heart and diaphragm, as well as the peripheral and central nervous system. Thus current research efforts into the elucidation of the molecular mechanisms underlying these genetic diseases are not only directed towards studying skeletal muscle necrosis but also investigate abnormalities of heart and brain dystrophin glycoprotein complexes in cardiomyopathy and brain deficiencies associated with muscular dystrophy. Furthermore, many isoforms of dystrophin and dystrophin associated components have been identified in various non-muscle tissues and their function(s) are mostly unknown. With respect to skeletal muscle fibers, the characterization of new dystrophin-associated proteins, such as dystrobrevin, sarcospan and the syntrophins, led to a modified model of the spatial configuration of the dystrophin-glycoprotein complex. However, it is generally accepted now that beta-dystroglycan forms the plasmalemma-spanning linkage between dystrophin and the laminin-binding protein alpha-dystroglycan and that this complex is associated with the sarcoglycan subcomplex of sarcolemmal glycoproteins. PMID- 9850731 TI - Inhibitory effect of theobromine on induction of angiogenesis and VEGF mRNA expression in v-raf transfectants of human urothelial cells HCV-29. AB - Neovascularisation plays a crucial role in solid tumor growth and metastasis formation. Our previous studies showed that theophylline and theobromine suppressed cutaneous neovascular reaction induced in mice by human blood leukocytes, and lung as well as ovarian cancer cells. Here, we investigated the in vivo effect of theobromine on angiogenic activity of human urothelial cell line HCV-29, v-raf transfected (mouse cutaneous assay), and the in vitro effect of this drug on VEGF, tPA, uPA and PAI mRNA expression in these cells (RT-PCR method). Theobromine suppressed angiogenesis induced in mice by HCV-29-v-raf cells, inhibited VEGF mRNA expression, and had no effect on transcription of uPA and tPA in these cells. HCV-29-v-raf transfectants do not display transcripts of PAI, in the presence or the absence of theobromine. PMID- 9850732 TI - Transforming growth factor-beta protection of cancer cells against tumor necrosis factor cytotoxicity is counteracted by hyaluronidase (review). AB - Numerous cancer cells, when exposed to transforming growth factor beta (TGF beta), become resistant to tumor necrosis factor (TNF) cytotoxicity. Pretreatment of L929 fibroblasts, for example, with TGF-beta isoforms (beta 1, beta 2 and beta 3) for at least 0.5-1 h results in resistance to TNF killing. TGF-beta 1 mediates the following sequential events in L929 cells: i) rapid induction of protein tyrosine-phosphorylation (< 30 min), ii) stimulation of protective protein synthesis and acquisition of TNF resistance (approximately 0.5-1 h), and iii) suppression of I kappa B-alpha expression (1-2 h). Two protective proteins induced by TGF-beta 1 are a 46 kDa extracellular matrix TNF-resistance triggering (TRT) protein and a putative transmembrane anti-apoptotic adhesion protein TIF2 (containing and RGD motif in the extracellular region). Both proteins enable L929 cells to resist TNF killing. Notably, testicular hyaluronidase increases TNF sensitivity in several types of cancer cells, counteracts TGF-beta-mediated TNF resistance, and suppresses TGF-beta 1 gene expression in L929 cells in a serum dependent manner. Moreover, hyaluronidase antagonizes TGF-beta-mediated inhibition of epithelial cell growth. Both TGF-beta and hyaluronidase are essential for the progression and invasiveness of breast, prostate and other cancers. Conceivably, a stage-dependent expression, as well as a balanced production, of these proteins is essential for cancer development and self protection against TNF cytotoxicity. PMID- 9850733 TI - Functional role of epoxyeicosatrienoic acids and their production in astrocytes: approaches for gene transfer and therapy (review). AB - The list of functions attributed to astrocytes in the brain is ever increasing. These cells contain cytochrome P450 enzymes that have recently demonstrated a number of exciting roles besides detoxification. The P450 monooxygenases can covert the substrate arachidonic acid to epoxyeicosatrienoic acids (EETs), metabolites that mediate vasodilation, mitogenesis, platelet aggregation, Ca2+ signaling and steroidogenesis. Integration of other physiological pathways present in astrocytes with P450 mediated EET formation has generated a number of interesting hypotheses to yield deeper insight into the role of astrocytes in the brain. In order to test these hypotheses as well as to enhance the benefits of EETs in astrocytes, we have used viral-mediated gene transfer and overexpression of one cytochrome P450 monooxygenase, 2C11, to engineer astrocytes for gene manipulation and possible gene therapy. This review outlines evidence for the presence of EETs in astrocytes, the function of EETs and the progress made with viral vectors expressing epoxygenase for gene manipulation in astrocytes. PMID- 9850734 TI - Honokiol induces apoptosis in human lymphoid leukemia Molt 4B cells. AB - The exposure of human lymphoid leukemia Molt 4B cells to honokiol led to both growth inhibition and the induction of apoptosis. Morphological change showing apoptotic bodies was observed in the cells treated with honokiol. The fragmentation by honokiol of DNA to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time dependent. These findings suggest that growth inhibition by honokiol of Molt 4B cells results from the induction of apoptosis in the cells. PMID- 9850735 TI - New medicinal treatment for severe gingivostomatitis. AB - We formulated de novo a poraprezinc-sodium alginate suspension (P-AG) as a specific treatment for severe gingivostomatitis and administered it to 15 patients who had developed such inflammation while on chemotherapy. Very high utility of P-AG was demonstrated and the response was classified as excellent in 10 patients and good in 5 patients. The mechanism of the beneficial effect of P AG in treatment of severe gingivostomatitis accompanied by hemorrhagic erosion and ulcers is considered to involve the mucosal protective effect, free radical scavenging activity and tissue repair promoting action of poraprezinc together with the hemostatic action of sodium alginate. PMID- 9850736 TI - Induction of hypothermia, hypoglycemia and hyperinsulinemia after acute leptin immunoneutralization in overnight fasted mice. AB - Acute immunoneutralization of circulating leptin, with an anti-leptin antibody, significantly reduced rectal temperature at 30 min and 75 min post-injection in overnight fasted and at 30 min in overnight fed mice, while no effects in metabolic and ponderal indicators were observed after antibody administration for 22 days. Furthermore, hyperinsulinemia and hypoglycemia were induced by passive immunization against leptin, being both influenced by the post-prandrial status. These experiments confirm through an indirect approach that leptin is involved in energy, but also in glucose homeostasis. PMID- 9850737 TI - Azidothymidine resistance of H9 human T-cell lymphoma cells is associated with decreased sensitivity to antitumor agents and inhibition of apoptosis. AB - Biology of HIV-1 associated neoplasias is modulated by viral and host factors. In addition the development of tumors and their response to therapy may be further influenced by long-term treatment of HIV-1 patients with nucleoside analogs such as AZT (3'-azido-3'deoxythymidine), ddI (2',3'-dideoxyinosine), ddC (2',3' dideoxycytidine), d4T (2',3'-didehydro-2'3'-dideoxythymidine), and 3TC [(-)-beta L-2',3'-dideoxy-3'-thiacytidine] alone or in combination. As these compounds can trigger mechanisms involved in chemoresistance, we tested whether prolonged in vitro treatment of H9 cells (T-cell lymphoma) with AZT alters sensitivity of lymphoma cells to antitumor agents used for AIDS-associated malignancies. H9 cells grown for more than two years in medium containing 250 microM AZT developed resistance to the toxic effects of AZT while retaining sensitivity for other nucleoside analogs including ddC or cytosine arabinoside (ARA-C). These cells designated H9rAZT250 were 2 to 10-fold less sensitive to the toxic effects of antitumor agents, including cisplatin (CDDP), vincristine (VCR), doxorubicin (DOX) and etoposide (VP-16), when compared with parental H9 cells. The resistance of H9rAZT250 cells to antitumor agents was associated with inhibition of apoptosis as demonstrated by ultrastructural investigations and DNA-fragmentation assay (ELISA). The expression of the antiapoptotic gene bcl-2 was increased in H9rAZT250 cells while expression of other genes involved in the regulation of apoptosis such as c-myc, p53 and Fas was not changed. These results demonstrate that prolonged in vitro treatment of H9 lymphoma cells with AZT results in the development of resistance to antitumor agents in association with inhibition of apoptosis and increased expression of bcl-2. Therefore AZT long-term treatment of some HIV-1 patients with malignancies may have affected behavior of tumor cells including response to therapy. PMID- 9850738 TI - Characterization of recombinant precursor proteins of the human seminal plasma sperm motility inhibitor synthesized in insect cells. AB - Human seminal plasma sperm motility inhibitor (SPMI) proteins which are exclusively secreted from seminal vesicles, inhibit sperm motility. It is secreted as biologically active 52 kDa and a mixture of 71 and 76 kDa precursor forms, which are identical to semenogelin-I and II (Sg-I and Sg-II), respectively. To understand the molecular mechanism underlying the inhibition of sperm motility by SPMI proteins, we expressed human Sg-I and Sg-II genes in insect cells using a baculovirus system. The baculoviruses expressing full-size Sg-I and Sg-II proteins that were N-terminally-tagged with a hexahistidine were selected, and were infected with Sf 21 cells. The Sg-I and Sg-II proteins were purified from infected cells by column chromatography using Ni-NTA resin 48 h after infection. The full-size Sg-I and Sg-II proteins were obtained in soluble forms. However, they tended to aggregate to form a gel, as expected from naturally occurring semenogelin. Both the purified recombinant Sg-I and Sg-II proteins showed strong SPMI activities with a complete inhibition of sperm motility at 60 units/mg, equivalent to the natural proteins. This production system that permits the generation of purified Sg-I and Sg-II proteins, as well as mutant derivatives, will be helpful for further study on male infertility. PMID- 9850739 TI - The feasibility of replacement therapy for inherited disorder of glycolysis: triosephosphate isomerase deficiency (review). AB - Triosephosphate isomerase (TPI, EC 5.3.1.1) is an ubiquitously expressed enzyme that catalyses the interconversion of dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate in the energy-generating glycolytic pathway. Inherited defects in the TPI gene are characterised biochemically by markedly reduced TPI enzyme activity in all tissues resulting in metabolic block in glycolysis, with accumulating DHAP particularly in red cells. Clinical TPI deficiency is a rare autosomal recessive multi-system disorder characterised by non-spherocytic haemolytic anaemia, recurrent infections, cardiomyopathy, severe and fatal neuromuscular dysfunctions. Reviews of current literature show that after 30 years since TPI deficiency was first described, the disease still remains without effective therapy. However, several potential therapeutic strategies exist for the treatment of inherited metabolic disorders such as TPI deficiency. Development of an effective therapy for TPI deficiency presents a fascinating and formidable challenge for basic laboratory and clinical research. The major aim of this overview is to discuss the current knowledge of TPI deficiency with special emphasis on research efforts directed towards reversing the metabolic effects of the disorder. PMID- 9850740 TI - Comparative epidemiology of cancers of the testis, lung, bladder and stomach with special reference to the possible implication of environmental hormones in the recent risk changes of the 4 neoplasia types. AB - The purpose of this study is to extract common features of an environmental hormone-oriented neoplasia by comparatively investigating the epidemiological characteristics of testicular cancer in Denmark with those of other cancers, of which the age-adjusted incidence rates (AAIRs) underwent remarkable increase or decrease in the time range of early 1960's to mid 1980's. Practically, the log transformed (log) AAIRs and the corresponding log ASIRs (age-specific incidence rates) were used in parallel to investigate the dynamic aspect of cancer risk changes in time and space. The present study includes cancers of the testis, lung, bladder and stomach as study subjects, and followed the chronological transition of cancer risk for each tumor type and for each population unit from early 1960's to mid 1980's. In space, the present study includes the data for 6 population units as follows: Denmark, Birmingham-England, the State of New York less New York city, Miyagi-Japan, Puerto Rico and Cali-Colombia. Since 3 neoplasias other than testicular cancer were associated with male predominance of cancer risk, 1st order regression analysis was applied to a set of 5 chronologically consecutive data of log AAIRs for a given tumor to comparatively investigate the sex discrimination of cancer risk for each of 3 neoplasia types. Results obtained are as follows: a) the ASIR profile of testicular cancer in Denmark (a high-risk country) was a composite of an adult type surge and an infant type surge (a product of in utero carcinogenic insult). Consistent ascension of both the adult type surge and the infant type surge of the ASIR profile was observed in parallel with the straight line increase of log AAIR of testicular cancer in Denmark. b) The ASIR profiles of testicular cancer for Miyagi-Japan (a low-risk country with steady increase of log AAIR) experienced new emergence of the infant type surge that was detectable in the profile for the years 1968-1971 and 1983-1987, but not in the profile for the years 1959-1960. c) The ASIR profiles of testicular cancer for Cali-Colombia (a low-risk country with no sign of risk increase) was free of the infant type surge throughout the study period. d) Temporary emergence of the infant type surge was experienced in cancers of the lung and bladder in early 1970's. In space, the infant type surge had preference of occurrence for a high-risk country to a low-risk country, and for a male population to a female population. e) Another feature of environmental hormone-oriented tumor was found in the recent risk decrease of gastric cancer of which the rate of risk decrease was distinct in Western countries and Japan, but not in Cali-Colombia. Puerto Rico was ranked as an in-between existence--a violation to the rules of Westernization effect for cancer risk. f) The relation between the male log AAIR and the male log AAIR less female log AAIR for a given tumor type was found to have a good fitness to the equilibrium model of which the members are destined to interact with each other under the law of mass action. The above mathematical strictness was found to be valid with all of cancers of the liver, skin, lung, bladder, stomach and esophagus--a finding to indicate that the relation between the changes of male cancer risk and that of female cancer risk in time and space is defined by the law of mass action regardless of the presence or absence of environmental hormone impact, and that no marker is available to detect possible implication of the environmental hormones with this system. g) The significance of the last finding in cancer etiology was discussed in the light of the steroid criminal hypothesis of human carcinogenesis in general. In conclusion, the recent risk changes of cancers of all the testis, lung, bladder and stomach in high-risk areas are in part to be explained in terms of the environmental hormone impact. PMID- 9850741 TI - Cox-2, iNOS and p53 as play-makers of tumor angiogenesis (review). AB - Cyclooxygenases (COXs) are key enzymes in the conversion of arachidonic acid to prostaglandins (PGs) and other eicosanoids. Nitric oxide synthase (NOS) is the enzyme that catalyzes the formation of nitric oxide (NO), a regulator of vascular permeability, from the guanidino nitrogen atom of L-arginine. Two isoforms of both enzymes occur: a constitutive one, Cox-1 and the inducible counterpart Cox 2; also NOS has a constitutive counterparts (cNOS) and an inducible form, called iNOS. The inducible isoforms of both enzymes are of maximum interest. It has been recently shown that cyclooxygenase-2 (Cox-2) is inducible by a variety of stimuli and that eicosanoids, mainly of the PGE2 species, are inducers of basic regulator of angiogenesis, including VEGF/VPF, bFGF, TGF-beta, PDGF, and endothelin-1. In addition, iNOS is inducible by Cox-2. p53 down-regulates the angiogenic process at various levels: it induces thrombospondin-1, a powerful antiangiogenic factor, down-regulates VEGF and NOS and, in addition, down-regulates hypoxia-induced angiogenesis, either inducing apoptosis or enhancing antiangiogenetic factors. It is noteworthy how important the p53 oncosuppressor is in the angiogenesis of solid tumor growth. Cox-2, iNOS and p53 are thus fundamental play-makers of the angiogenic process: they are discussed in detail and a tentative hierarchical cascade is proposed. PMID- 9850742 TI - Patterns of the immunohistochemical expression of melanoma-associated antigens and density of CD45R0+ activated T lymphocytes and L1-protein positive macrophages in primary cutaneous melanomas. AB - There is ample evidence for spontaneous antimelanoma immune reactivity mediated by melanocyte-differentiation-antigens (MDAs). Our aim was to determine whether MDA immunoreactivity is associated with increased tumour-infiltrating lymphocytes (TIL) and macrophages (TIM). A retrospective study was conducted in 30 medium and high grade primary cutaneous melanomas (PCM) as identified by CART-analysis. All of the cases had developed clinical evidence for metastasis within 3 years following surgical excision of the PCM. We used immunohistochemistry and computerized image analysis to quantify MDAs positive cells (Melan A/MART-1, gp100/Pmel 17/HMB45, tyrosinase), CD45R0-positive TIL and LI-protein-positive TIM. A stochastic relationship was present between the MDA immuno-reactivities and the densities in TIL and TIM. An inverse relationship was yielded between TIL and TIM. No specific pattern of PCM immunoreactivity for MDAs, TIL and TIM was found to predict metastases. PMID- 9850743 TI - Vasoactive intestinal peptide and epidermal growth factor: co-mitogens or inhibitors of keratinocyte proliferation in vitro? AB - Vasoactive intestinal peptide (VIP) is a neuropeptide with a broad range of biological activities in various tissues. Interactions of VIP and epidermal growth factor (EGF) are of particular interest for dermatology. They may be either co-mitogenic or inhibitory. HaCaT keratinocytes cultivated under serum free conditions in vitro have been used to investigate the interactions of VIP and EGF. EGF was found to induce cell growth, whereas preincubation with VIP inhibited EGF-induced proliferation in a dose-dependent manner. Maximum growth inhibition was 46% (p < 0.01) at a VIP concentration of 10(-7) M. EGF-induced growth is mediated by tyrosine kinase (TK). Therefore we studied the effect of VIP on TK activity. Cells were incubated with VIP (10(-13)-10(-7) M) for 48 h and stimulated with EGF at a final concentration of 500 ng/ml. SDS-PAGE and Western blot with the antibody RC20H against TK were performed. We found a dose dependent decrease of EGF receptor TK activity. At VIP concentration of 10(-7) M a residual TK activity of 65% was detected. To investigate the possibly involved signal transduction pathways, we performed inhibition experiments with wortmannin, pertussis toxin, 2'5'diacylglycerol and adenosine-3':5'-mono-phosphorothioate. However, none of the inhibitors was effective in abolishing growth inhibition by VIP. VIP was shown to be growth inhibitory for human keratinocytes. The data suggest that EGF receptor TK is involved in signal transduction of VIP. Thus TK activity is a possible common target of both EGF- and VIP-induced cellular responses. PMID- 9850744 TI - Emergency physician's diagnosis of stroke subtype. An accuracy study. AB - OBJECTIVE: To evaluate the accuracy of clinical unstructured and structured diagnosis of acute stroke subtypes--cerebral haemorrhage (CH), cerebral infarction (CI), subarachnoid haemorrhage (SAH). METHODS: Sixty consecutive patients with acute stroke admitted to the Emergency Ward of a Brazilian University Hospital were examined by emergency physicians and computerised tomography (CT). We also compared it (physician's unstructured diagnosis) to two published clinical scoring systems (structured diagnosis--Guy's Hospital and Siriraj Hospital) applied to three other populations--regarding the operational characteristics of the tests. RESULTS: In our personal data, among 9 variables that could discriminate CH and CI, three have statistically significant difference (p < 0.05): headache (p = 0.0002) and vomiting (p = 0.02) occurred more frequently in CH patients, but previous stroke in those with CI (p = 0.04). Unstructured diagnosis proved valid for SAH, with a +LHR = 39.7; and to a smaller degree for CI (-LHR = 0.1). However, it exhibited low sensitivity for the diagnosis of CH. Structured tests (Guy's Hospital and Siriraj Hospital) also failed to confidently diagnose stroke subtypes, especially CH. CONCLUSIONS: Both clinical diagnosis (made by emergency physicians) and the available diagnostic tests fail to confidently discriminate CH and CI. PMID- 9850745 TI - Outcome of 337 intracranial aneurysms patients operated in a public hospital. AB - Many recent series of surgery for intracranial aneurysms have been based on experience of developed countries with great resources and a state of art health care. The purpose of the current study is to correlate the outcome of patients operated for intracranial aneurysms, reported from intensive high technology neurosurgical centers with the results of low technology, environment, where we practice. Between January 1986 and December 1996, 337 patients with intracranial aneurysms were operated on at the Servidores do Estado Hospital. We retrospectively reviewed the medical and radiologic records and compared the outcome of this group with other series derived from developed countries. The overall mortality of this series was 6.9%. Of the 313 good grades surgical patients, the mortality was 4.7% and the successful results were obtained in 88.8% individuals. We conclude that patients harboring intracranial aneurysms can be satisfactory handled in less developed nations, if a meticulous intraoperative technique is employed, even though sophisticated technology and equipment are not available. PMID- 9850746 TI - Preliminary results in surgery of Parkinson's disease. AB - The authors present the preliminary results of 20 patients selected to be operated on between January 1996 and April 1997. These patients presented one of the present indications for stereotactic posteroventral pallidotomy (PVP), such as: rigidity, akinesia/bradykinesia, gait dysfunction, drug induced dyskinesias and tremor. Every patient of this protocol was evaluated by: UPDRS score, Schwab and England scale, Hoehn and Yahr Staging Scale before and after surgery. The results in 3 months showed a remarkable improvement after PVP (P < 0.01) in all functional assessments, except for facial expression, speech and posture. The morbidity was 5%. 5 patients (25%) who were in Hoehn and Yahr 5 underwent a bilateral simultaneous PVP. In 5 patients (25%), who had tremor, during the PVP, VIM thalamotomy was added. These preliminary results, suggest that PVP is highly effective for PD symptoms. PMID- 9850747 TI - Non increased neuron-specific enolase concentration in cerebrospinal fluid during first febrile seizures and a year follow-up in pediatric patients. AB - Febrile seizures are the commonest acute neurological disorder of early childhood. Studies suggested that febrile seizures are previous acute events from a more serious neurological problem. Due to neuron-specific enolase is generally accepted as a marker for neuropathological processes in the brain, 16 pediatric patients were studied during their first seizures and a year after it. Neuron specific enolase in cerebrospinal fluid and blood were analysed by an immune enzyme assay. Non pathological neuron-specific enolase values were obtained in both periods in the group of patients. There were no significative differences when paired series statistics test was performed with 95% of confidence. Neuron specific enolase appears not to be a marker for febrile seizures because its concentration not be increased in cerebrospinal fluid in this group of patients. PMID- 9850748 TI - [Glutaric aciduria type 1: phenotypic variability. Report of 6 patients]. AB - We report six patients with glutaric aciduria type 1 in four families. The patients had marked clinical variability, even within families. Three of the patients studied were normal until the onset of neurologic abnormalities, that presented as an encephalitis-like illness in the first year of age. One patient had an early and important developmental delay, but never suffered an encephalopathic crisis. Two patients have intellectual preservation; one of them has a mild tremor and choreoathetosis since the first year of age, and the other had only two afebrile seizures in infancy and no other neurologic signs. Three patients are severely handicapped, with a severe dystonic-dyskinetic disorder and unable to even sit. All the six patients have macrocephaly and in all the computed tomography showed enlarged CSF spaces and sulcal separation over the frontal and temporal lobes. Urine organic acids study of all patients showed large quantities of glutaric acid. PMID- 9850749 TI - [Migraine in childhood: difficulties in diagnosis]. AB - The study of chronical headache in children, mainly migraine, has deserved special attention in the last years. The publications point out their epidemiological importance and are unanimous to affirm that the diagnosis made do not correspond to clinical reality. They emphasize that one of the difficulties that justify this diversity would be inadequate diagnosis criteria. This study analyses a group of pediatric patients complaining about headache and reaches results that support this impression. PMID- 9850750 TI - [Headache incidence in a hospital community]. AB - The purpose was to describe the main features of headache incidence in a hospital community, its frequency and the most requested medical investigation. Due to the stressful work environment, hospital is considered to hold a high-risk population. Interviews and questionnaires were utilized. Of a 1006 files, which were randomly filled out, 987 could be analyzed. Of all, 38.5% were from headache sufferers. By using a table of pain symptoms taken from the International Headache Society classification as a pattern, headaches were assigned as migraine, tension-type and other. The mean age was 31.18 and the frequency in females was higher than in males, at any type. Family occurrence in first-degree relatives was 76.8%. Frontal location, medium intensity and pulsation were the most described features. Stress was the most frequently mentioned trigger factor. A physician was consulted only by 41.3%. Cranium X-ray was the most frequently requested exam. PMID- 9850751 TI - [Use of chlorpromazine in the treatment of headache at an emergency service]. AB - Therapeutic measures, with intravenous chlorpromazine, taken during acute headache are evaluated in fourteen patients at the emergency room in Santa Casa de Sao Paulo. Four patients had the diagnosis of migraine with aura and five patients migraine without aura. Four patients had diagnosis of chronic daily headache with intermittent and superimposed migrainous events. Finally one patient had the diagnosis of chronic paroxysmal hemicrania. All patients were diagnosed according to International Headache Society criteria. The intravenous chlorpromazine dose used was 0.7 mg/Kg diluted in 5% glucose solution and the dose never exceeded 50 mg. The time of drug administration was never less than 60 minutes. The results were considered excellent in all cases. Some patients presented side effects, particularly orthostatic hypotension, always moderate and transitory. This study has clearly demonstrated that intravenous chlorpromazine (0.7 mg/Kg) was highly effective in terminating episodes of primary headache. PMID- 9850753 TI - [Neurocysticercosis: contribution of autopsies in the consolidation of mandatory notification in Ribeirao Preto-SP,Brazil]. AB - Neurocysticercosis is a serious public health problem in several countries of Asia, Africa and Latin America. The objective of the present study is to present autopsy findings of neurocysticercosis in the Pathology Division of the University Hospital, Medical School of Ribeirao Preto, University of Sao Paulo, from 1992 to 1997. Neurocysticercosis was detected in 38 (1.5%) of 2522 autopsies. In twenty two (57.9%) of these, the diagnosis was made post-mortem, while 16 (42.1%) had a previous diagnosis of cysticercosis based on laboratory and imaging studies. Reviewing the medical records, we found that 21 patients (55.2%) were neurologically asymptomatic, while the various clinical manifestations in the others were in accordance with the literature. The high frequency of asymptomatic individuals detected only post-mortem indicates the importance of autopsies as an additional source of data to consolidate the compulsory notification of cysticercosis. Considering the results of this study, the new coefficient of prevalence of cysticercosis in Ribeirao Preto is 67 cases/100,000 inhabitants. PMID- 9850752 TI - [Chronic brain edema in neurocysticercosis]. AB - In a retrospective study, we report the clinical characteristics of chronic brain oedema (CBO) in 34 patients with neurocysticercosis (NCC) who presented diffuse brain oedema upon computed tomography (CT) as a common point. All patients received dextrochlorpheniramine, and, 94.1% of them also albendazole. A predominance of females (73.5%) aged 11-40 years (92.3%) was seen. Headache occurred in 94.1% of patients, nausea/vomiting in 47.1%, epileptic manifestations in 41.1% and psychiatric disorders in 38.2%. Hyperreflexia was obtained in 82.3% of patients, papilloedema in 58.8% and normal neurological examination in 11.8%. The CT scan of the brain showed an association of oedema with calcifications in 61.8% of cases. The cerebrospinal fluid (CSF) pressure was higher before than after treatment (p < 0.05). At the last visit, 79.4% of patients were currently asymptomatic (57.1% of them without medication). It is pointed out that CBO may be an antigenic manifestation in NCC without the concomitant presence of parasitic cysts and may represent an additional clinical condition associated with benign intracranial hypertension. PMID- 9850754 TI - [Ulnar nerve palsy in leprosy without skin changes: biopsy of the superficial branch of the ulnar nerve in the hand]. AB - Leprosy is one of the most common diseases of peripheral nerves in the world. In Brazil it is particularly frequent, being a major health problem. In tuberculoid leprosy the ulnar nerve is the most common affected nerve. Sometimes there are no skin changes. In these cases in spite of nerve thickening only the nerve biopsy is capable to make a specific diagnosis. We performed a biopsy in the dorsal sensory branch of the ulnar nerve in the hand in 17 patients with ulnar palsy with thickening of the nerve in the elbow, without skin changes. The pathological findings consisted mainly of: loss of fibers (14 cases), inflammatory infiltration (13), fibrosis (12), demyelination and remyelination (9), presence of granuloma (6) and presence of bacilli (5 cases). We conclude that in case of ulnar nerve palsy in leprosy without skin changes, the biopsy of the dorsal sensory branch of this nerve in the hand is a good procedure for the diagnosis of leprosy. PMID- 9850755 TI - [Epilepsies with midline projected foci: study with quantitative and topographic EEG (brain mapping)]. AB - We report a topographic and quantitative EEG (EEGQT) study of four patients with analogic EEG diagnostics of midline foci. The new study with EEGQT offered an increased definition of the electrical source with advantages in foci localization. These findings should be confirmed with studies including a greater number of patients. PMID- 9850756 TI - [Computerized tomography in agenesis of the corpus callosum: findings in 27 cases]. AB - We describe a retrospective study of 27 cases of complete agenesis of the corpus callosum examined by CT and not associated to schizencephaly, holoprosencephaly and Dandy-Walker complex. Partial agenesis is also not included in the present study. The imaging findings are correlated to sex, age and symptoms. PMID- 9850757 TI - [Mini mental state examination and the diagnosis of dementia in Brazil]. AB - The diagnosis of dementia is based on the presence of memory deficits and decline of other cognitive functions. Many scales have been designed to aid the clinician in the assessment of at risk subjects, such as the elderly. The Mini-Mental State Examination (MMSE) is the most widely used of such scales, although its use as an aid to the diagnosis of dementia has not as yet been studied in a Brazilian sample of patients. The current study was designed with 2 main aims: (1) to determine the best cut-off point of the MMSE for the diagnosis of dementia in a sample of elderly subjects assessed in a mental health outpatient unit and; (2) evaluate the impact of age and schooling on MMSE scores. Two hundred and eleven subjects aged 60 or over assessed at the Mental Health Outpatient Unit for the elderly at "Santa Casa de Sao Paulo" between February 1997 and February 1998 were included in the study. They were assessed with the SRQ-20, MMSE, and a clinical interview for the diagnosis of dementia according to the ICD-10. Seventy patients received the diagnosis of dementia. The MMSE cut-off point of 23/24 (cases/non cases) was associated with a sensitivity of 84.3% and specificity of 60.3%. MMSE scores were associated with age (r = -0.41, p < 0.001) and schooling (F = 12.69, p < 0.001). Analysis of covariance taking age into account showed that MMSE scores were significantly lower among those with no formal education (F = 10.51, p < 0.001). A cut-off point of 19/20 on the MMSE was associated with sensitivity of 80.0% and specificity of 70.9% for the diagnosis of dementia in this particular group. Subjects with previous school history were better classified to the diagnosis of dementia with the cut-off point of 23/24: sensitivity of 77.8% and specificity of 75.4%. The evaluation of elderly subjects with the MMSE should take education into account, and different cut-off points should be used accordingly as a guideline to the diagnosis of dementia. PMID- 9850758 TI - [Hemostatic and structural effects of the lyophilized cellulose sponge]. AB - Hemostatic effects of oxidized cellulose (Surgicel) are well known. Based on a possible similar effect of a sponge obtained after lyophilization of biosynthetic cellulose, two different experimental studies were planned. Phase I-Pieces of cellulose sponge were inserted into small provoked cortical wounds of twelve dogs. The time elapsed to obtain bloodstill after cortical damage and application of cellulose was observed in every dog, searching to detect any possible hemostatic effect of the material. The animals were sacrificed after 7, 30 and 90 days. An average time of 1 minute was elapsed until bleeding control was achieved. No clinical adverse effect was noticed. Microscopy showed histiocytic and mild foreign body reaction at 7 days, which diminished at 30 days. Almost no reaction surrounded the implant at 90 days. Lyophilized cellulose has a peculiar eosinophilic appearance, composed by thin irregular filaments which diminished their thickness with the time. At 90 days only sparse irregular cellulose filaments could be detected. Phase II-Small equal sponge fragments were inserted in the liver of twelve rats and observed 7, 30 and 90 days. At autopsy, small peritoneal adhesions were noticed at 30 and 90 days. Microscopy showed intense histioplasmocytic and foreign body reaction in all animals mainly at 7 days. In two animals, refringent intracellular cellulose particles were evident inside giant foreign body cells after 90 days. This fact evidences that cellulose can be reabsorbed by phagocytic phenomena when implanted in mammalians. A comparative group with other hemostatic material and the same method must be done to clarify the issue of hemostatic effects of this membrane. PMID- 9850760 TI - Epidural hematoma after ventriculoperitoneal shunt surgery. Report of two cases. AB - Ventriculoperitoneal shunt operations represent the most used choice for treating hydrocephalus, although some related complications have been reported. Due to its rarity, potential dangers, and mortality rate, we present two cases of epidural hematoma following ventriculoperitoneal shunt, discussing its pathophysiology and prophylaxis. PMID- 9850759 TI - [Isolated fourth ventricle: report of 2 cases]. AB - Two cases of isolated fourth ventricle are reported, the first due to cerebellar haemorrhage, and the second due to congenital hydrocephalus with multiple shunt revisions and Dandy-Walker cyst. In our opinion, there are two basic treatment for isolated forth ventricle. The direct approach to the fourth ventricle is indicated when there is presence of an intraventricular cyst. The fourth ventricular shunting, independent of the supratentorial shunt, is the best treatments for patients with an isolated fourth ventricle without the presence of a cyst. PMID- 9850761 TI - Evaluation of two infants with myotonic dystrophy by the McFie's diagram from the results of WISC. AB - In this paper the authors disclose the result of a research carried out on two brothers whose parents were first cousins, being the gene transmitted by the father. The psychological test Wechsler Intelligence Scale of Children (WISC) was used in two occasions in order to assess the verbal and non-verbal skills. FRM and IRM were nine and eleven-year-old respectively, in the first examination, being the former thirteen and the latter fifteen-year-old on the second one. A comparison between the McFie's diagram and the WISC scores was made: the McFie's diagram showed the impairment severity in each cortical lobe when the left hemisphere was compared with the right one. The McFie's diagram was made from WISC's scores: the McFie's diagram showed the impairment severity in each cortical lobe when the left hemisphere was compared with the right one. On the second examination the performance was worse than in the first, mainly in the non verbal aspects. The IRM's diagram showed a reduction in the right frontal and parietal lobes. In the FRM's diagram a reduction in the left frontal, temporal and parietal lobes, and also, in the right parietal lobe was found. The visual spatial constructive aspects showed greatest impairment in this result. PMID- 9850762 TI - [X-linked recessive bulbospinal muscular atrophy (Kennedy's disease). A family study]. AB - Kennedy's disease is a rare type of motor neuron disease with a sex-linked recessive trait. DNA studies show a mutation at the androgen receptor gene on the long arm of X chromosome (Xq 11-12) with expanded CAG triplets (more than 347 repeats). We present three patients and one carrier among ten patients of a four generation family with clinical phenotype of the disease. The patients' ages ranged from 50 to 60 years with symptomatology usually beginning around 30 years of age. Patients had gynecomastia, testicular atrophy, muscular weakness, fasciculation, amyotrophy, absent deep tendon reflexes and postural tremor. PCR techniques of DNA analysis showed expanded size of CAG repeats on Xq 11-12 in all the three patients and in the carrier asymptomatic woman. This is the first Brazilian family with genetic molecular diagnosis of Kennedy's disease. This disease must be included in the differential diagnosis of motor neuron disease since it has a distinct prognosis and genetic counseling is mandatory to the carriers. PMID- 9850763 TI - Folie a deux dissociative disorder in prepubertal children. Report of two cases with EEGs. AB - A case of folie a deux dissociative (dissociative hysteria) disorder in an 8 and 12 year-old sister and brother is presented. Illnesses of this type are very rare and there is little medical literature on this subject. Our patients, almost simultaneously, abruptly had complete loss of memory, disorientation, loss of awareness about who they were, and much anxiety, which lasted about 15 hours. Both patients were physically well and no abnormalities were found on physical examination, routines laboratory tests and EEG studies. Speculations about the emotional and interpersonal causes of this illness in these two patients are given. PMID- 9850764 TI - Kleine-Levin syndrome. Clinical course, polysomnography and multiple sleep latency test. Case report. AB - A case of Kleine-Levin syndrome, with chronic severe periodic hypersomnia is described in a 17-year-old female. The first episode started when she was 15 years old. The episodes were characterized by periodic hypersomnia accompanied by hyperphagia, lasting 5 days, and repeating at 28 to 60 day intervals. The severity of hypersomnia prevented her from attending school activities. Outside the hypersomnia periods, she was asymptomatic. EEG, brain computerized tomography and brain nuclear magnetic resonance were normal; all-night polysomnography, Multiple Sleep Latency Test (MSLT) and Epworth Sleepiness Scale (ESS) were within normal limits. During the period of hypersomnolence, polysomnography showed short sleep latency and short REM latency. MSLT mean sleep latency was 1.8 min; and REM period was present in one subtest; the ESS was markedly elevated. PMID- 9850765 TI - Jactatio capitis nocturna with persistence in adulthood. Case report. AB - Rhythmic movement disorder, also known as jactatio capitis nocturna, is an infancy and childhood sleep-related disorder characterized by repetitive movements occurring immediately prior to sleep onset and sustained into light sleep. We report a 19-year-old man with a history of headbanging and repetitive bodyrocking since infancy, occurring on a daily basis at sleep onset. He was born a premature baby but psychomotor milestones were unremarkable. Physical and neurological diagnostic workups were unremarkable. A hospital-based sleep study showed: total sleep time: 178 min; sleep efficiency index 35.8; sleep latency 65 min; REM latency 189 min. There were no respiratory events and head movements occurred at 4/min during wakefulness, stages 1 and 2 NREM sleep. No tonic or phasic electromyographic abnormalities were recorded during REM sleep. A clinical diagnosis of rhythmic movement disorder was performed on the basis of the clinical and sleep studies data. Clonazepam (0.5 mg/day) and midazolam (15 mg/day) yielded no clinical improvement. Imipramine (10 mg/day) produced good clinical outcome. In summary, we report a RMD case with atypical clinical and therapeutical features. PMID- 9850766 TI - Hemorrhagic cerebral metastasis as a first manifestation of a hepatocellular carcinoma. Case report. AB - We report herein a rare instance in which a patient presented with a hemorrhagic cerebral metastasis as the initial manifestation of a hepatocellular carcinoma (HCC). A few cases of cerebral metastasis from HCC have been reported in the literature, mainly from eastern countries. This is the first report from South America of a cerebral metastasis from hepatocellular carcinoma. PMID- 9850767 TI - Lipoblastic meningioma. Case report. AB - We describe the case of a patient presenting a right parietal mass lesion with an heterogeneous aspect on computed tomography, with hyperdense contrast uptake areas and hypodense areas with fat density. The unusual aspect of the lesion prevented preoperative and intraoperative diagnosis. The final histopathological examination revealed a meningothelial neoplasia with adipose differentiation, characterizing a lipoblastic meningioma. PMID- 9850768 TI - [Germinoma of the basal ganglia and thalamus with brain stem invasion: case report]. AB - We report an unusual case of germinoma arising from the basal ganglia and thalamus with brain stem invasion, with emphasis on computed tomography and magnetic resonance findings. Diagnosis was confirmed by histopathologic examination. Early detection of this tumor is important due to its potential response to treatment. PMID- 9850769 TI - [Tuberous sclerosis: case report with histopathological and ultrastructural study]. AB - Tuberous sclerosis complex is a group of autosomal disorders characterized by hamartomas and benign neoplastic lesions that invariably affect the central nervous system. We report a case of tuberous sclerosis that is the first presenting ultrastructural findings of this phacomatosis in the Latin American literature. The patient was a 2 year old girl presenting West syndrome non responsive to the clinical treatment with vigabatrin, trileptal and clonazepan, and undergoing left frontal lobectomy. The histopathological and ultrastructural findings were compatible with tuberous sclerosis. These results may help to further understand this controversial phacomatosis, warning to the clinical presentation as West syndrome. PMID- 9850770 TI - [Neurophysiology of pain in tentorial irritation: description of a case secondary to medulloblastoma]. AB - We report the case of a 32-year-old woman who complained of dull and compressing occipital pain, with unilateral radiation to the left frontal and supraorbital areas (of the sickening type). The radiological, clinical, neurosurgical and neuropathologic investigation disclosed a medulloblastoma bulging and tickening the tentorium cerebelli. This case shows how mechanical stimuli of the structures innervated by the tentorial nerve can cause pain with characteristics of trigeminal and cervical involvement. PMID- 9850771 TI - [Neuro-urological findings in Williams syndrome: report of a case]. AB - The Williams syndrome is a relatively rare disease with characteristic facial appearance, mental retardation, growth deficiency, cardiovascular anomalies, hypercalcemia and multiple organic dysfunctions. However, the urological findings of this syndrome (positive in up to 40% of patients) have not been frequently discussed. We present the case of a 6 year-old white girl with this diagnosis and a 3-year history of urinary incontinence. The investigation revealed bladder diverticula and detrusor hyperactivity, which was successfully treated with oxibutimin. We stress the importance of urological investigation, describe the main findings and discuss the pathophysiology and management, which significantly improves the quality of life of these children. PMID- 9850772 TI - [Treatment of Alzheimer' disease: critical evaluation of the use of anticholinesterase]. AB - Alzheimer's disease (AD) is a neurogenerative disorder that had no effective treatment until a few years ago. Many studies published during the past 20 years showed that the disease is associated with the disruption of brain systems that use acetilcholine. These findings shaped the development of the cholinergic treatment strategies for AD. The cholinesterase inhibitors were the first class of drugs to show positive results in double-blind placebo controlled studies for the treatment of AD. This paper reviews efficacy and safety studies of tacrine, donepezil, rivastigmine, and metrifonate in patients with AD. Factors that may interfere with patients' response to treatment are also discussed, as well as the limitations associated with the use of this class of drugs. PMID- 9850773 TI - [Technical recommendations for the electroencephalogram (EEG) recording in suspected brain death]. AB - Brazilian Clinical Neurophysiology Society guidelines and pertaining comments concerning electroencephalogram (EEG) recording in suspected brain death are presented. EEG is not intended as a substitute, rather as a complement to neurologic evaluation. PMID- 9850774 TI - Posterior labral injury in contact athletes. AB - Nine athletes (seven football offensive linemen, one defensive lineman, and one lacrosse player) were found at arthroscopy to have posterior labral detachment from the glenoid. In our series, this lesion is specific to contact athletes who engage their opponents with arms in front of the body. All patients had pain with bench pressing and while participating in their sport, diminishing their ability to play effectively. Conservative measures were ineffective in relieving their symptoms. Examination under anesthesia revealed symmetric glenohumeral translation bilaterally, without evidence of posterior instability. Treatment consisted of glenoid rim abradement and posterior labral repair with a bioabsorbable tack. All patients returned to complete at least one full season of contact sports and weightlifting without pain (minimum follow-up, > or = 2 years). Although many injuries leading to subluxation of the glenohumeral joint occur when an unanticipated force is applied, contact athletes ready their shoulder muscles in anticipation of impact with opponents. This leads to a compressive force at the glenohumeral joint. We hypothesize that, in combination with a posteriorly directed force at impact, the resultant vector is a shearing force to the posterior labrum and articular surface. Repeated exposure leads to posterior labral detachment without capsular injury. Posterior labral reattachment provides consistently good results, allowing the athlete to return to competition. PMID- 9850775 TI - Injury of the suprascapular nerve at the spinoglenoid notch. The natural history of infraspinatus atrophy in volleyball players. AB - From 1985 to 1996, we observed 38 cases of isolated atrophy of the infraspinatus muscle in athletes; all were involved in volleyball at a competitive level. There were 20 men and 18 women with a mean age of 26 years (range, 15 to 27). At the time of the first examination, 35 of these athletes had no pain and were treated with exercises to strengthen the external rotators. The remaining three patients underwent surgery because of pain at the posterior aspect of the shoulder. Sixteen of the 35 players treated nonoperatively were reviewed at a mean follow up of 5.5 years (range, 3 to 10). Thirteen were still involved in volleyball and three had retired symptom-free at the end of their careers. On physical examination, atrophy of the infraspinatus muscle was unchanged in all cases. The patients treated surgically were reviewed at a mean follow-up of 2 years. All of them were able to play volleyball at their preinjury levels, but one had pain at the anterior aspect of the shoulder after strenuous activity. Physical examination showed a notable reduction of the atrophy in one patient. Entrapment of the suprascapular nerve at the spinoglenoid notch is a usually painless syndrome that is frequently observed in volleyball players. Surgical treatment is indicated in the rare cases of painful neuropathies after careful patient selection. PMID- 9850776 TI - The development and evaluation of a disease-specific quality of life measurement tool for shoulder instability. The Western Ontario Shoulder Instability Index (WOSI). AB - The purpose of this study was to develop a valid, reliable, and responsive disease-specific quality of life measurement tool for patients with shoulder instability. Development included 1) identification of a specific patient population; 2) generation of issues specific to the "disease" ("items") from reviewing the literature, interviewing health caregivers, and interviewing patients representing all demographics, disease type and severity, and treatments; 3) item reduction using patient-generated frequency-importance products and correlation matrices; and 4) pretesting the prototype instrument on 2 groups of 10 patients. The final instrument, the Western Ontario Shoulder Instability Index, has 21 items representing 4 domains. The instrument attributes (validity, reliability, and responsiveness) were evaluated. Construct validation demonstrated that this index correlated predictably with other measures. Reliability was very high at 2 weeks and 3 months, and the index was more responsive (sensitive to change) than five other shoulder measurement tools (the Disabilities of the Arm, Shoulder and Hand scale; The American Shoulder and Elbow Surgeons Standardized Shoulder Assessment Form; the UCLA Shoulder Rating Scale; the Constant Score; and the Rowe Rating Scale), a global health instrument (the SF12), and range of motion. Since the patient's perception of changes in health status is the most important indicator of the success of a treatment, we suggest that this measurement tool be used as the primary outcome measure to evaluate treatments in this patient population, although it can also be used for monitoring patients' progress in clinical practice. PMID- 9850777 TI - Conscious neurosensory mapping of the internal structures of the human knee without intraarticular anesthesia. AB - The conscious neurosensory characteristics of the internal components of the human knee were documented by instrumented arthroscopic palpation without intraarticular anesthesia. With only local anesthesia injected at the portal sites, the first author (SFD) had both knees inspected arthroscopically. Subjectively, he graded the sensation from no sensation (0) to severe pain (4), with a modifier of either accurate spatial localization (A) or poor spatial localization (B). The nature of the intraarticular sensation was variable, ranging from 0 on the patellar articular cartilage to 4A on the anterior synovium, fat pad, and joint capsule. The sensation arising from the cruciate ligaments ranged from 1 to 2B in the midportion, and from 3 to 4B at the insertion sites. The sensation from the meniscal cartilages ranged from 1B on the inner rim to 3B near the capsular margin. Innervation of most intraarticular components of the knee is probably crucial for tissue homeostasis. Failure of current intraarticular soft tissue reconstructions of the knee may be due, in part, to the lack of neurosensory restoration. Research studies of the knee designed to delineate factors that restore neurosensory characteristics of the musculoskeletal system may lead to techniques that result in true restoration of joint homeostasis and function. PMID- 9850778 TI - Functional knee brace effects during walking in patients with anterior cruciate ligament reconstruction. AB - The purpose of this study was to compare lower extremity joint kinematics and kinetics during walking with and without a functional knee brace in patients with recent anterior cruciate ligament reconstructions. Seven volunteers walked at 1.26 m/s with and without one of two functional knee braces 3 weeks after surgery. Eleven uninjured subjects were also tested as a control group. Video and ground-reaction data were collected and combined with inverse dynamics to estimate the joint positions, moments, and powers during the stance phase. Patients with ligament reconstructions were more erect with the brace, using 19% less knee flexion compared with walking without the brace. Areas under the internal extensor moment curve (angular impulse) and power curve (work) at the hip increased 40% and 44%, respectively, while walking with the brace. Extensor angular impulse decreased 41% at the knee while using the brace, and plantar flexor angular impulse and work increased 21% and 30%, respectively, at the ankle. While walking with the brace, the patients still had different kinematics, moments, and powers than the control subjects. The reduced extensor moment at the knee in the braced condition indicated that the load on the recently reconstructed ligament was reduced and that the brace protected the ligament during the stance phase of walking. We concluded that functional knee braces may be one means of developing neuromuscular adaptations during gait after anterior cruciate ligament reconstruction surgery. PMID- 9850779 TI - Surgical repair of chronic complete hamstring tendon rupture in the adult patient. AB - Complete rupture of the hamstring tendons in the adult is a rare injury. This report discusses complete rupture of the hamstring tendons in nine patients treated by late operative repair. All patients were referred from outside centers for a second opinion after failed nonoperative treatment. The diagnosis was made quite easily on clinical grounds and was confirmed at surgery. Surgical treatment in all cases consisted of reattachment of the hamstring tendons to the origin on the ischium, and in all cases it was necessary to perform neurolysis of the sciatic nerve. Good results were achieved in all cases, at follow-up all patients were satisfied with the surgery. PMID- 9850780 TI - Early full weightbearing and functional treatment after surgical repair of acute achilles tendon rupture. AB - We prospectively evaluated the clinical outcomes of 20 patients (mean age, 42.8 years) with early full weightbearing and functional treatment after surgical repair of acute Achilles tendon rupture according to a prospective intra- and postoperative protocol. All patients underwent open repair using a Kessler-type suture and simple apposition sutures. The postoperative regimen included a plantigrade splint for 24 hours and 6 weeks of early full weightbearing in a removable walker. All patients were evaluated with clinical and ultrasound examination and according to a new scoring system at 3, 6, and 12 months after repair. After 3 months, the score averaged 73 of 100 points; after 6 months, 86; and after 1 year, 94. All patients reached the same level of sports activities as preoperatively and demonstrated no significant difference in ankle mobility and isokinetic strength. There were no reruptures. One patient had a deep venous thrombosis 3 weeks after the operation after having prematurely stopped thromboprophylaxis. We believe that early careful ankle mobilization and full weightbearing in a removable walker after primary Achilles tendon repair does not increase the risk of rerupture. An accelerated rehabilitation program improves early foot function with excellent recovery of plantar flexion strength and amplitude. PMID- 9850781 TI - Early active motion and weightbearing after cross-stitch achilles tendon repair. AB - Twenty-two closed Achilles tendon ruptures caused by sports injuries in 22 patients (average age, 37.6 years) were repaired with Kirschmayer core suture and cross-stitch epitenon suture, and early active ankle motion with weightbearing was implemented after surgery. This study was undertaken to evaluate the effectiveness of the repair technique and rehabilitation protocol by assessing clinical results and magnetic resonance imaging findings. The follow-up period averaged 24.6 months. Twenty of the tendons (91%) healed without rerupture, and two tendons (9%) suffered a partial rerupture at 23 and 56 days, respectively. Active ankle extension reached from the minus range to 0 degree in an average of 9.7 days, and ankle motion recovered to normal in an average of 6.0 weeks. Full weightbearing without heel raising became possible in an average of 16.4 days, and heel raising with both legs became possible in an average of 7.3 weeks. The patients returned to full sports activity in 13.1 weeks. The interval until the area of high-intensity signal at the tendon repair site on T2-weighted magnetic resonance imaging scans became intermediate-intensity signal averaged 6.9 weeks, and the tendon repair site became low-intensity signal in an average of 12.6 weeks, demonstrating excellent tendon healing. Treatment employing Kirschmayer core suture and cross-stitch epitenon suture may help athletes return to sports activity in a shorter period than that allowed by previous methods of repair for Achilles tendon ruptures. PMID- 9850782 TI - The effects of overuse combined with intrinsic or extrinsic alterations in an animal model of rotator cuff tendinosis. AB - An in vivo animal model was used to evaluate overuse and overuse plus intrinsic tendon injury or extrinsic tendon compression in the development of rotator cuff injury. Forty-four male Sprague-Dawley rats were divided into groups of 22. Each left shoulder received an intrinsic or extrinsic injury plus overuse (treadmill running), and each right shoulder received only overuse. Eleven rats from each group were sacrificed at 4 and 8 weeks. Supraspinatus tendons were evaluated histologically or geometrically and biomechanically. Ten rats constituted a cage activity control group. Both supraspinatus tendons of the experimental groups had increases in cellularity and collagen disorganization and changes in cell shape compared with control tendons. Tendons with injury plus overuse exhibited a worse histologic grade than those with overuse alone. The cross-sectional area of both supraspinatus tendons of the experimental rats was significantly more than in control tendons. The area of the injury plus overuse tendons was increased on average compared with overuse-alone tendons. Biomechanically, the tissue moduli of overuse/intrinsic injury tendons at 4 weeks and those of the overuse/extrinsic injury tendons at 8 weeks were significantly lower than in control tendons. Tissue moduli of the overuse/injury tendons were significantly lower than in the overuse-alone tendons at 8 weeks. This study demonstrated that damage to the supraspinatus tendon can be caused by overuse and intrinsic injury, overuse and extrinsic compression, and overuse alone. PMID- 9850783 TI - The thermal effect of monopolar radiofrequency energy on the properties of joint capsule. An in vivo histologic study using a sheep model. AB - The purpose of this in vivo study was to analyze the short-term tissue response of joint capsule to monopolar radiofrequency energy and to compare the effects of five power settings at 65 degrees C on heat distribution in joint capsule. In 12 mature Hampshire sheep, the medial and lateral aspects of both stifles were treated with monopolar radiofrequency energy under arthroscopic control in a single uniform pass to the synovial surface. The radiofrequency generator power settings were 0, 10, 15, 20, 25, and 30 watts (N = 8/group). The electrode tip temperature was 65 degrees C. Histologic analysis at 7 days after surgery revealed thermal damage of capsule at all radiofrequency power settings. The lesion's cross-sectional area, depth, vascularity, and inflammation were commensurate with radiofrequency power. Tissue damage was indicated by variable inflammatory cell infiltration, fusion of collagen, pyknosis of fibroblasts, myonecrosis, and vascular thrombosis, whereas synovial hyperplasia, fibroblast proliferation, and rowing of sarcolemmal nuclei demonstrated regenerative processes. This study revealed that radiofrequency power settings and heat loss through lavage solution play a significant role in heat distribution and morphologic alterations in joint capsule after arthroscopic application of monopolar radiofrequency energy. PMID- 9850784 TI - Comparison of initial fixation strength between biodegradable and metallic interference screws and a press-fit fixation technique in a porcine model. AB - The objective of this study was to evaluate the initial fixation strength of a biodegradable interference screw compared with press-fit fixation and a titanium interference screw in anterior cruciate ligament reconstruction using a bone patellar tendon-bone graft. Porcine lower limbs were used. The specimens underwent 500 loading cycles between 60 and 250 N. This corresponds to loads in the graft during aggressive rehabilitation. Thereafter, intact specimens were loaded to failure. Failure mode was defined by visual analysis. Under cyclic loads none of the interference screw fixations failed. In the press-fit group (angle between load axis and tunnel axis 80 degrees), five specimens failed. The mean maximal load to failure was 945 N (+/- 87) for the titanium screw, 797 N (+/ 60) for the biodegradable screw, and 708 N (+/- 211) for the five press-fit specimens that did not fail during cyclic loading. With respect to primary fixation strength, biodegradable screws are a reasonable alternative to titanium interference screws. The press-fit fixation did not provide a secure fixation in all cases. Five press-fit specimens failed under cyclic loads comparable with those seen under conditions of accelerated rehabilitation. PMID- 9850785 TI - Postexercise increase in nitric oxide in football players with muscle cramps. AB - Nitric oxide, a free radical inter- and intracellular messenger molecule, is important in exercise physiology. This study tested the hypothesis that serum nitric oxide concentrations change after strenuous exercise with severe generalized muscle cramps. The study group consisted of 77 professional football players in preseason training. All players' concentrations of serum nitrite and of other serum chemicals were determined during their preseason evaluations and compared with the concentrations in 40 serum samples taken from 25 of those same players who required intravenous rehydration for severe generalized muscle cramps after a training session. Player weight and percentage of body fat were significantly higher in players who received intravenous fluids than in players who did not. The serum of players requiring intravenous hydration showed evidence of skeletal muscle breakdown (increases in lactate dehydrogenase, creatinine phosphokinase, aspartate aminotransferase, and alanine aminotransferase) and of dehydration (elevations in protein, blood urea nitrogen, and cholesterol). The major finding, however, was a nearly 300% increase in serum nitrite concentrations in players requiring rehydration. There were no correlations between concentrations of nitrate and of any of the other serum chemicals. These data support the hypothesis that large amounts of nitric oxide are synthesized in professional football players after strenuous exercise with severe muscle cramps. The study design did not allow us to determine whether this increase in nitric oxide was due to exercise or muscle cramps or both, but it does provide a basis for evaluating these relationships. PMID- 9850786 TI - Dynamic impact response of human cadaveric forearms using a wrist brace. AB - The purpose of this study was to compare the dynamic impact response of braced and unbraced cadaveric wrists using a commercially available wrist guard. Twelve arms were harvested from six cadavers. Each pair of forearms, one with and one without a brace, were impacted using a modified guillotine-type drop fixture placed over a force platform. Using a piece-wise linear regression analysis, we identified four phases of dynamic loading in the vertical force profile before fracture. These phases included an initial linear loading phase starting at impact, followed by a nonlinear phase, a second rapid linear loading phase, and a final nonlinear loading phase to failure. Three transition points were identified that defined the boundaries of the linear loading phases. Vertical force and impulse were significantly higher (P < 0.01) at each transition point and at failure in all braced specimens compared with unbraced specimens. However, the most noticeable differences were found during the initial two loading phases. Time to each transition point and to failure was not significantly different (P > 0.27) between the braced and unbraced wrists. The results of this study differ from those obtained under more quasistatic loading conditions. Dynamic impact testing suggests that wrist guards may have a prophylactic effect during low energy dynamic impact situations. PMID- 9850787 TI - Biomechanical performance of Bankart repairs in a human cadaveric shoulder model. AB - The objective of this study was to develop a method to evaluate the biomechanical performance of Bankart repairs in a human cadaveric shoulder in a clinically relevant orientation. Twenty fresh-frozen human cadaveric shoulder girdles were used to compare the biomechanical performance of intact anteroinferior capsulolabral complexes with the biomechanical performance of three Bankart lesion reconstruction techniques. Repairs were performed on surgically created Bankart lesions. Evaluations were performed with the shoulders in glenohumeral abduction and external rotation. The repair techniques employed interosseous sutures, Mitek GII suture anchors, or Acufex T-Fix devices. The suture material used in all repairs was No. 2 Ti-Cron. The biomechanical performance of the three reconstruction techniques did not differ, but each was significantly inferior compared with that of the intact shoulder samples. The interosseous repairs failed by suture pullout through soft tissue. Repairs in the Mitek GII group failed by pullout of the suture anchors, suture breakage, or pullout of the suture through soft tissue. Repairs in the T-Fix group failed by pullout of the suture through soft tissue or failure of the polymer portion of the T-Fix suture. PMID- 9850788 TI - Multiplanar analysis of acromion morphology. AB - To more completely describe acromion morphology and its relationship to impingement syndrome, we performed three-dimensional magnetic resonance imaging (N = 111) or computed tomography (N = 27) on 132 symptomatic shoulders. The mean patient age was 46.2 years (range, 14 to 86). Four parameters were evaluated: the angle of anterior slope of the acromion in the midsagittal and lateral-sagittal planes, lateral acromial angulation in the coronal plane, and the presence or absence of medial encroachment in the acromioclavicular joint. Twenty-five asymptomatic age-matched shoulders were used as controls. All imaging data were combined because no significant differences existed between the two imaging techniques. The mean acromion angle was 19.4 degrees in the midsagittal plane and 20 degrees in the lateral-sagittal plane. In the coronal plane, 97 (73%) acromions were neutral and 35 (27%) were downward sloping. Medial encroachment was present in 31 (24%) shoulders. Age distribution from the 2nd to 8th decade demonstrated a consistent and gradual transition from a flat acromion in the younger decades to a more hooked acromion in the older decades that was significant in both the midsagittal and lateral-sagittal planes. Furthermore, a greater percentage of patients were found to have downward angulating acromions with increasing age. Ninety-eight patients (74%) had stage II or III impingement. Of these shoulders, 39 (40%) had type I acromions, 51 (52%) type II, and 8 (8%) type III. Twenty-eight of 33 acromions with coronal lateral downward sloping had impingement, and all 31 shoulders with medial encroachment had impingement. PMID- 9850789 TI - Sagittal plane translation of the knee during stair walking. Comparison of healthy and anterior cruciate ligament--deficient subjects. AB - With an electrogoniometer system, we made bilateral measurements of the maximal sagittal plane anterior-posterior knee translations in 15 healthy subjects (controls) and 14 patients with arthroscopically confirmed unilateral anterior cruciate ligament deficiency during two types of ascents and descents (straight and side). In both groups, during the ascent cycle the tibia moved anteriorly in relation to the femur, whereas during the descent cycle it moved posteriorly. There was wide individual variation in maximal translation in both the control and anterior cruciate ligament-deficient groups (range, 1 to 12 mm; mean, 7 mm). The maximal translations were similar in both groups (P > 0.05), but they occurred at a significantly smaller flexion angle in the injured knees (38 degrees +/- 8 degrees) than in the control and non-injured knees (44 degrees +/- 8 degrees) (P < 0.05). The translation during step ascent and descent did not differ between the injured and control knees. These findings indicate that patients with anterior cruciate ligament injuries are able to control abnormal anterior translation during normal activity. PMID- 9850790 TI - Avulsion injury to the conjoined tendons of the latissimus dorsi and teres major muscles. PMID- 9850791 TI - Straddle pelvic stress fracture in a female marathon runner. A case report. PMID- 9850793 TI - The strain behavior of the anterior cruciate ligament during squatting and active flexion-extension--a comparison of an open and a closed kinetic chain exercise. PMID- 9850792 TI - Articular cartilage lesions of the knee. PMID- 9850794 TI - Pattern of protein retention in growing boars of different breeds, and estimation of maximum protein retention. AB - Protein and energy metabolism in boars of different breeds, 10 each of Hampshire, Duroc and Danish Landrace was measured in balance and respiration experiments by means of indirect calorimetry in an open-air circulation system. Measurements were performed in four periods (Period I-IV) covering the body weight range from 25 to 100 kg. In order to achieve maximum protein retention (RP) a daily intake of digestible protein > 12 g/kg0.75 and metabolisable energy > 1100 kJ/kg0.75 was assumed to be necessary. Protein retention of Danish Landrace boars was inferior to that of Hampshire and Duroc boars in Periods III and IV, and therefore, 55 measurements on Hampshire and Duroc boars fulfilling the chosen criteria for digested protein and ME intake were used for calculation of maximum protein retention, giving the following significant quadratic relationship: RP [g/d] = 11.43.W0.75-0.144.W1.50 (n = 55, RSD = 15.2, CV = 9.2%, R2 = 0.851) with a summit of 227 g/d at 135 kg BW. In Period I, when BW was below 30 kg, 12 measurements fulfilled the chosen criterion for digested protein but not for ME, and these data were used comparatively. Protein retention of boars with a low ME intake in Period I was significantly below that of boars with a high ME intake (93 g/d vs. 107 g/d; P = 0.02). In summary, the present data have shown that boars of high genetic potential have capacity for maximum protein retention of about 230 g/d, and that there was a significant quadratic relationship between protein retention and metabolic body weight, indicating that maximum protein retention was not reached until 135 kg BW. Differences in capacity for protein retention were recorded between boars of different breeds, with Duroc and Hampshire boars being superior to Danish Landrace boars. Additionally, the crucial importance of a sufficient ME supply early in the growth period was underscored by a lower protein accretion rate of boars given a daily ME supply below 1100 kJ ME/kg0.75 at an approximate BW of 25 kg. PMID- 9850795 TI - [Energy metabolism in laying hens of different body weight genotypes]. AB - Energy metabolism and some performance parameters were investigated in laying hens of 3 different body weight-genotypes: 6 x 7 (normal-sized, crossbred from normal-sized male and female lines, group 1), 47 x 38 (dwarf-sized, breeding from a dwarf-sized male line and a normal-sized female line, group 2) and 44 x 47 (dwarf-sized, breeding from dwarf-sized male and female lines, group 3). Energy balance was measured by indirect calorimetry through C- and N-balances in 12 animals of each group during 10 consecutive days at production peak, within the period between the 27th and the 37th week of age. Hens were caged individually at 23 degrees C environmental temperature and fed ad libitum on a laying mash. The mean body weight in the dwarf-sized groups 2 and 3 was 32% lower than in the normal-sized group 1 during the energy balance period. The daily gross energy intake in group 2 and 3 was decreased by 33 and 34%, respectively. There were no significant differences in digestibility and metabolizability of gross energy between the groups. The energy requirements for maintenance [kJ ME/kg0.75.d] derived from the energy balances were lower by 4% (P > 0.05) and 14% (P < 0.05) in the dwarf-sized groups 2 and 3 as compared with the normal-sized group 1, when equal coefficients of partial efficiency of metabolizable energy utilization for energy retention in body and eggs are assumed for the 3 body-mass genotypes. There were no relevant differences in body composition at the end of the energy balance periods as well as in egg composition between the 3 experimental groups. PMID- 9850796 TI - Influence of dietary concentrate to forage ratio on the development of rumen mucosa in calves. AB - Effects of structural and non-structural carbohydrates on the development of rumen fermentation and ruminal mucosa in calves were examined during the weaning period. Barley/soybean meal (SBM) group was fed a concentrate starting from 2 weeks of age, whereas alfalfa group received a mixture of concentrate and alfalfa hay in which the proportion of the latter was gradually increased from 20% to 70% between weeks 2 and 9 of age. The total volatile fatty acid concentration in rumen fluid of calves increased with age, but at 9 weeks there were no significant differences between the two diets (barley/SBM group 153 mmol/l, alfalfa group 150 mmol/l). Rumen papillae at 9 weeks of age, as compared to 6 weeks of age, were longer and fewer in number per square centimetre mucosa, with larger cut surface. This resulted in a higher surface of papillae per square centimetre mucosa at 9 weeks (barley/SBM group 286 mm2/cm2, alfalfa group 245 mm2/cm2) than at 6 weeks of age (barley/SBM group 217 mm2/cm2, alfalfa group 198 mm2/cm2). At 9 weeks of age, the pH (barley/SBM 5.0, alfalfa 5.7), the acetate to propionate ratio (barley/SBM 2.2, alfalfa 3.2) as well as the length of the papillae in the ventral ruminal sac (barley/SBM 1.96 mm, alfalfa 2.37 mm) were increased in the alfalfa group when compared to the barley/SBM group (P < 0.1). In the former group, the proportion of butyrate revealed significantly increased values at 4 and 6 weeks of age. In animals of the barley/SBM group at 9 weeks of age, characteristic protrusions with proliferated thick epithelium occurred on the papillae and increased the surface for absorption. On the epithelium (Stratum corneum) desquamating cells with parakeratosis could be observed. In the alfalfa group the papillae of the ventral ruminal sac were longer, without protrusions. The morphotypes of the adhering rumen microflora differed between the groups. It can be concluded that feeding greater amounts of non-structural carbohydrates increases the surface for absorption of the rumen epithelium in calves. The absence of hyperkeratosis and rumenitis in the barley/SBM group indicated that there is no reason to limit high starch diets in the early weaning period of calves. PMID- 9850797 TI - Studies on the mode of action of non-starch-polysaccharides (NSP) degrading enzymes in vitro. 1. Communication: effects on the fractions of NSP. AB - An in vitro model using the model substance wheat bran as NSP source was developed to study the effects of NSP degrading enzymes (xylanase and beta glucanase) on NSP fractions. The in vitro model simulates the conditions (pH, dry matter, temperature and transit time) in the fore sections of the porcine gastrointestinal tract. By neglecting endogenous enzymes the sole effect of the added enzymes could be investigated. By application of the enzyme complex, a significant degradation of the insoluble NSP fraction was observed in all sections of the porcine gastro-intestinal tract simulated by the in vitro model. A strong dose dependent effect was determined. Compared to the control, the arabinose/xylose-ratio generally increased but differences were found between the sections simulated. Considering the arabinoxylans in the soluble NSP fraction, the effect of enzyme supplementation was reflected by a change in the arabinose and xylose contents. In comparison to the control, an increase in the amounts of arabinose and xylose was observed at low enzyme doses applied. However, with increasing enzyme activities the amount of arabinose and xylose declined. The in vitro model is considered as a useful tool for preliminary tests to estimate the effects of NSP-degrading enzymes on NSP fractions in the fore sections of the porcine gastro-intestinal tract. PMID- 9850798 TI - The use of an in vitro cellulase enzyme technique for predicting the digestibility of tropical feeds and fodders. AB - An attempt has been made to find out if the organic matter digestibility of tropical feeds and fodders can be predicted from the results of an in vitro cellulase enzyme technique. The in vivo digestibility coefficient of 27 tropical feeds and fodders was determined in wethers. The enzyme insoluble organic matter content of these feeds was determined using a cellulase enzyme technique. The indigestible OM (g/kg feed DM) in vivo (y) was plotted against enzyme insoluble organic matter (g/kg DM) (x). The equation is y = 60 + 0.817x, R2 = 0.95; RSD = 32 g/kg DM. Equations for predicting DOM (g/kg DM) and digestibility of OM (%) are calculated. The testing of the equation on an independent data base of 40 individual feeds and fodders showed that the validity of the equation is fairly general. PMID- 9850799 TI - The comparison of in vitro fermentation kinetics estimated by three different methods. AB - Three different methods for the estimation of in vitro fermentation kinetics are compared. The glass syringe, flow gasometer and pressure transducer methods were used for measurement of gas production. The rumen fluid from fistulated Merino sheep mixed with McDougall's buffer (1:1) was used as an inoculum and added at an amount of 35 ml into the fermentation vessels containing 0.25 g of meadow hay. The total gas produced was recorded after 12, 24, 36, 48, 60, 72 and 96 h of incubation. Hay dry matter degradability was the same with all three methods and achieved 56.5 to 58.2%. Total volatile fatty acids were significantly lower with the pressure transducer method than with the syringe and flow gasometer method. Lower values of mol% of butyrate and valerate obtained with the flow gasometer and pressure transducer methods in comparison with the syringe were also observed. Total gas production estimated by the flow gasometer method was lower than that stated by the two other methods. With regard to precision of the used methods syringe method was the best followed by the pressure transducer and flow gasometer method. It can be concluded that in spite of some limitations the pressure transducer method used in this experiment can be regarded as suitable for total gas estimation in in vitro rumen fermentation experiments. PMID- 9850800 TI - [The effect of size reduction of corn silage on feed intake, milk production and milk composition of cows]. AB - In three experiments (E1, E2, E3) maize silage of different physical structure and of different stage of maturity at harvest were fed to 24 (E1), 36 (E2) or 28 (E3) dairy cows. The cows were fed individually over an experimental period of five or six weeks. The maize silages had a mean DM content of 28% (E1), 32% (E2) or 36% (E3). At the stage of harvest, the stovers and the cobs had a mean DM content of < 22% (E1, E2) or 27% (E3), 40% (E1), 46% (E2) or 57% (E3), respectively. The maize was harvested with a chopping length of 4 and 8 mm (E1, E3) and of 6 and 8 mm (E2), without corn cracking (E1) or with and without corn cracking (E2, E3). The daily feed ration consisted of ad libitum offered maize silage, 1.7 kg DM hay, soya bean meal (E2, E3) and concentrate. The different chopping length of 4 mm, 6 mm or 8 mm had no effect on the maize silage intake in E1 and E2. In E3 the daily maize silage intake increased by about 1.2 kg DM per cow at a chopping length of 4 mm in comparison to 8 mm, whereas only the treatment with the combination of 4 mm chopping length and corn cracking showed a significant increase in DMI. The corn cracking improved the milk yield significantly (E2) or in a tendency (E3) at 2.0 kg (E2) or at 1.6 kg (E2), while the variation of chopping length had no effect on milk yield. The different physical structure did not influence the milk fat content with mean values of 4.65% (E1), 4.15% (E2) and 4.10% (E3), respectively. The milk protein content decreased in E2 feeding maize silage with a chopping length of 8 mm and corn cracking; but in E1 and E3 no effect was seen on protein content with mean values of 3.66% (E1) or 3.51% (E2). PMID- 9850801 TI - Research note: investigation on the metabolism of glycerol in the rumen of bulls. AB - Two bulls, each fitted with rumen and duodenal cannulas, received (in addition to a hay-grain diet) twice daily an infusion of 200 g glycerol into the rumen over a period of six days. During this preliminary in vivo investigation, the influence of a glycerol application on the rumen environment over a six-day adaptation period was examined. Samples of rumen fluid were collected daily, two hours after glycerol infusion. An additional 15N-urea application into the rumen was given on days 1 (without glycerol infusion), 3 and 7 (with glycerol infusion). Extra samples of rumen fluid and blood plasma (from puncture of vena jungularis) were taken through the 12th hour following urea application. Rumen fluid pH was reduced due to glycerol intake from 6.3 (day 1, without glycerol) to 5.4 by day 7. Molar proportion of acetic acid to propionic acid decreased from 3.5 (day 1) to 2.1 (days 6 and 7). Average glycerol disappearance rate from the rumen was 4.7 gl-1 h-1 for the first hour. Only small amounts of glycerol could be detected in the duodenal digesta. Blood plasma glycerol content was significantly higher after glycerol application (0.061 mmol l-1 vs. 0.019 mmol l-1). The incorporation of 15N into the rumen bacteria and the proportion of bacterial N (as percent of TCA-precipitable N in the rumen fluid) were lower after glycerol influsion. These results, coupled with the lower concentration of iso-acids (isobutyric and isovaleric acids) in the rumen fluid, indicate that the high amount of glycerol infusion (10% of DMI) reduced protein metabolism of rumen bacteria throughout the experimental period. PMID- 9850802 TI - Research note: effect of duodenal--infused unsaturated fatty acids on dairy milk composition. AB - Investigations were conducted to assess the effect of duodenal infusions of oleic and linoleic acids both on milk composition and fatty acid patterns of milk fat. The investigations were carried out in two trials, I and II, each with 3 cows, fitted with a ruminal and a duodenal cannula. 56 g of oleic acid (Trial I) and 46 g linoleic acid (Trial II) were infused daily into the duodenum over a period of 7 days, while no application of fatty acids acted as a control in each case. An increase in milk fat content was observed in both trials whereas the content of protein and lactose in milk remained unaffected. In both trials the milk oleic acid and linoleic acid concentrations increased. In trial I C18:1 increased from 14.8 to 18.7%. In trial II C18:2 increased from 2.5 to 5.8% of total lipids. It was estimated that 84.8% of the infused C18:1 and 86.5% of C18:2 were transferred into milk fat. PMID- 9850803 TI - Is there a connection between car accidents, near accidents, and type A drivers? AB - The authors examined Type A behavior pattern, which has been investigated primarily as a risk factor for coronary heart disease, as a risk factor for car accidents and near accidents. Type A behavior pattern, which is characterized by excessive impatience, competitiveness, hostility, and time pressure, was assessed by means of the Videotaped Structured Interview. One hundred thirty-five Swedish car drivers (66 men and 69 women) were studied: 78 Type A and 58 Type B (that is, not having, Type A behavior). Time pressure was significantly associated with near accidents when age, sex, annual mileage, and urban driving were controlled in a multivariate model. PMID- 9850804 TI - Mind machines: a controlled study on the effects of electromagnetic and optic acoustic stimulation on general well-being, electrodermal activity, and exceptional psychological experiences. AB - Mind machines are said to induce relaxation and an altered state of consciousness. The influence of optical-acoustic (OAS) and electrical stimulation (ES) mind machines on general well-being, electrodermal activity, and altered states of consciousness was investigated and compared with pure rest and a placebo. Thirty participants underwent 4 conditions in random sequence: rest, placebo stimulation, OAS, and ES. Statistical evaluations before and after the experimental stimulations showed differences in general well-being and electrodermal activity measures but no between-conditions effect. ES and OAS produced significantly more visionary experiences and fear of ego dissolution than rest and placebo. Differences in mystical experiences were found between ES and OAS. Mind machines do not have a specific effect on general well-being and physiological relaxation, although they may produce unusual psychological experiences; people with psychiatric illnesses should not use such devices. PMID- 9850805 TI - How daughters of women with breast cancer cope with the threat of the illness. AB - Coping strategies and attitudes toward medical treatment of 45 daughters of women who had had breast cancer were compared with strategies (sense of coherence) and attitudes on a health opinion survey of 51 women with no such family history who were referred to a breast health clinic in northern Israel. The daughters of women with breast cancer ranked lower on sense of coherence than the women with no such history. They also were more actively involved in the medical setting and requested more medical information than women whose mothers did not have breast cancer. The age of the daughter at the time her mother's cancer was diagnosed predicted her level of coherence and attitudes toward treatment. PMID- 9850806 TI - Religion and hypertension: testing alternative explanations among immigrants. AB - A number of researchers have found that religious commitment has a salutary effect on blood pressure levels. Levin and Vanderpool's review of several of these studies led them to offer some methodological, epistemological, and conceptual criticisms of this body of literature, and they advanced various hypotheses offering possible alternative explanations for the putative salutary effect of religion on blood pressure. In this article, the effect of religious commitment on hypertension among a sample of immigrants is examined with these hypotheses and criticisms in mind. Even when the majority of Levin and Vanderpool's criticisms are addressed, the religion effect remains. PMID- 9850807 TI - Late neurocognitive changes from neurological damage following coronary bypass surgery. AB - A detailed case study with neuropsychological data over a 26-year period was used to explore the relationship between neurocognitive vulnerability preoperatively and subsequent neurocognitive decline identified several years postoperatively. Guidelines regarding the importance of neuropsychological assessment of intelligence, attention, memory, language, and visual-spatial planning and organizational skills are provided. Such evaluations clarify postoperative treatment planning because rehabilitation of cardiac patients with premorbid neurocognitive deficits poses special rehabilitation problems. With a detailed neurologic history as part of the preoperative evaluation, healthcare providers can identify acute and subtle risk factors for postoperative neurologic syndromes. This may lead to interventions designed to provide increased patient and family support. PMID- 9850809 TI - Cobalamin metabolism in chronic myelogenous leukemia. PMID- 9850808 TI - Body fat, fat distribution, and psychosocial factors among patients with Type 2 diabetes mellitus. AB - Diabetes, a risk factor for cardiovascular disease, requires lifestyle modifications (diet, exercise, weight loss). The relations between body mass index, waist-hip ratio (WHR), and psychosocial indicators, such as affect and stress, among 302 diabetic patients from a clinic and a neighborhood health center were analyzed. Data included stress and mood scale responses, body size (height, weight, and WHR) and potential confounders (physical activity, energy intake, and diabetes duration). In univariate analyses, body mass index was positively associated with stress and inversely associated with positive affect only in women. Multiple regression analyses indicated that stress was associated with body mass index and negative mood was associated with the WHR. The findings suggested that stress and affect may be important correlates of body fat among women with Type 2 diabetes, leading to more complications. Healthcare providers can help women with Type 2 diabetes lose weight and lower the risk of cardiovascular disease by recognizing and helping them deal with these psychosocial issues. PMID- 9850810 TI - Prognosis and risk factors for bronchial asthma. PMID- 9850811 TI - Gestational diabetes mellitus and subsequent development of overt diabetes mellitus. AB - GDM develops in 1-3% of all pregnancies. Women with GDM are characterized by a relatively diminished insulin secretion coupled with a pregnancy-induced insulin resistance primary located in skeletal muscle tissue. The cellular background for this insulin resistance is not known. The binding of insulin to its receptor and the subsequent activation of the insulin receptor tyrosine kinase have significant importance for the cellular effect of insulin. Thus, the pathogenesis to the insulin resistance was studied by investigating insulin receptor binding and tyrosine kinase activity in skeletal muscle biopsies from women with GDM and pregnant controls. No major abnormalities were found in GDM wherefore it is likely that the insulin resistance is caused by intracellular defects distal to the activation of the tyrosine kinase. Glucose tolerance returns to normal postpartum in the majority of women with GDM. However, previous studies, in populations quite different from a Danish population, have shown that women with previous GDM have a high risk of developing overt diabetes mellitus later in life. Hence, we aimed to investigate the prognosis of women with previous GDM with respect to subsequent development of diabetes and also to identify predictive factors for the development of overt diabets in these women. A follow up study of diet treated GDM women diagnosed during 1978 to 1985 at the Rigshospital, Copenhagen was performed. Glucose tolerance was evaluated in 241 women (81% of the GDM population) 2-11 years after pregnancy. Abnormal glucose tolerance was found in 34.4% of the women (3.7% IDDM, 13.7% NIDDM, 17% IGT) in contrast to a control group where none had diabetes and 5.3% had IGT. Logistic regression analysis identified the following independent risk factors for later development of diabetes: a high fasting glucose level at diagnosis of GDM, a delivery more than 3 weeks before term, and an abnormal OGTT 2 months postpartum. Low insulin secretion at diagnosis of GDM was also an independent risk factor. The presence of ICA and GAD-autoantibodies in pregnancy was associated with later development of IDDM. In another study the following techniques: hyperinsulinaemic euglycaemic clamp, indirect calorimetry and tritiated glucose infusion were used to evaluate insulin sensitivity in glucose tolerant nonobese women with previous GDM and controls. A decreased insulin sensitivity due to a decreased non oxidative glucose metabolism in skeletal muscle was found in women with previous GDM. Hence, the activity of three key enzymes in intracellular glucose metabolism (GS, HK and PFK) was studied in skeletal muscle biopsies obtained in the basal state and after 3 h hyperinsulinaemia, with the aim to identify the cellular defects causing the decreased insulin sensitivity. However, no abnormalities in enzyme activity was found. The same group of previous GDM women had a relatively reduced insulin secretion evaluated by the IVGTT. A longitudinal study of 91 GDM women showed a relatively reduced insulin secretion to oral glucose in pregnancy, postpartum as well as 5-11 years later. Thus the present review has shown that even nonobese glucose tolerant women with previous GDM are characterized by the metabolic profile of NIDDM i.e. insulin resistance and impaired insulin secretion. Hence, the combination of this finding together with the significantly increased risk for development of diabetes indicates that all women with previous GDM should have a regular assessment of their glucose tolerance in the years after pregnancy. The first OGTT should be performed around 2 months postpartum in order to diagnose women already diabetic and to identify women with the highest risk for later development of overt diabetes. Women with previous GDM comprise a target group for future intervention trials with the aim to prevent or delay development of NIDDM and IDDM. PMID- 9850812 TI - Maintenance chemotherapy of acute lymphoblastic leukemia in children. PMID- 9850813 TI - Endoscopic sphincterotomy for common bile duct stones in younger patients. AB - BACKGROUND AND STUDY AIMS: Most follow-up studies of patients with common bile duct stones treated with endoscopic sphincterotomy include older patients. Therefore, the aim of the present study was to do a medium-term follow-up of patients under the age of 60 years, who underwent endoscopic sphincterotomy and removal of common bile duct stones. METHODS: Eighty-six (68 women) consecutive patients less than 60 years of age (median age 47 years) were evaluated. The median observation time after endoscopic sphincterotomy was 27.1 months. Complete follow-up was accomplished in 80 patients. RESULTS: Twenty-five patients had undergone cholecystectomy prior to the endoscopic sphincterotomy. Sixty-one patients were discharged after endoscopic sphincterotomy with gallbladder left in situ. Cholecystectomy was not needed in 27 patients, of whom 23 had gallbladder stones. In 13 of 61 patients subsequent elective cholecystectomy was planned at the time of endoscopic sphincterotomy. Recurrent attacks of biliary colic required elective cholecystectomy in another thirteen patients. Acute cholecystectomy was undertaken in eight patients due to acute cholecystitis (n = 4) or biliary colic (n = 4). Seven of eight acute cholecystectomies were performed within 152 days after the endoscopic sphincterotomy. The overall median time until cholecystectomy was 51 days (25-75 percentiles, 23-103 days). CONCLUSIONS: One third of the patients had been cholecystectomized prior to the endoscopic treatment of their common bile duct stones. Nearly half of the young patients discharged after endoscopic sphincterotomy for common bile duct stones with the gallbladder left in situ at the time of endoscopic sphincterotomy managed well without any need for cholecystectomy. PMID- 9850814 TI - [Subthreshold depressive disorders: description and importance for secondary prevention in psychiatry]. AB - "Subsyndromic" or "subthreshold" mood disorders belong to the category of mood disorders. Because newly studied, few informations are available up to date. The Appendix B of the DSM IV introduces six categories of research criteria which characterize these disorders and give us the thread of our study. From Hippocrate to contemporary specialists, many authors reported mild forms of mood disorders, including the Kraepelin or the psychoanalyst authors views. Dysthymic Disorder, Minor Depressive Disorder and Brief Recurrent Depressive Disorder are different categories of subthreshold unipolar disorders. During their course, these disorders overlap each other and with major mood disorders. Many studies, carried out in primary care practice, pointed out the severe impairment in social functioning, experienced by these patients. We propose a review of "Minor Depressive Disorders", focusing on some points: definitions, epidemiologic studies, "functional impact" of this kind of disorders, comorbidity and therapeutical considerations. Prevalence of suicide is extensive in non major depressive disorders. We discuss interest of "subsyndromic concept" aiming at the prevention of major mood disorders. Moreover, this concept leads to a new clinical approach in the care of mood disorders and provides new fields for psychopathological research. PMID- 9850815 TI - [Validation of a questionnaire of physical sensations in a French-Canadian population]. AB - The objectives of this study are three-fold. The first objective is to present the aspects of reliability and validity of a self-reported questionnaire that evaluates a specific facet of Panic Disorder with Agoraphobia (PDA) for the francophone population of Quebec, mainly the fear of fear. The Body Sensations Questionnaire (17), translated as the Questionnaire des Sensations Physiques (QSP) evaluates the degree to which individuals fear somatic symptoms commonly associated with panic. A second objective is to determine the factorial validity of the QSP and to investigate the dimensionality of the construct as assessed by the questionnaire. The third objective is to establish a reference database for the agoraphobic (N = 141 and N = 70) and the non-clinical population (N = 223). Results suggest that both questionnaires have similar internal consistency, temporal stability, construct validity, and discriminative validity. While the factorial solution of the present study uncovered a three-factor model (somatic, cardiac, and psychosensorial), which points to the multidimensional nature of the construct evaluated by the QSP, it greatly differs from the one obtained by Arrindel's study (5). Nevertheless, the extraction of the three principal components is supported by the results of the screen test and by two different implementations of parallel analysis, i.e., the computation of random eigen values and the use of an interpolation table of eigen values. Finally, it can be concluded that the psychometric properties of the French-Canadian translation of the BSQ are indications that it is an appropriate instrument to use in clinical research and in clinical practice to assess the fear of bodily sensations associated with the fear of fear. PMID- 9850816 TI - [Self-assessment questionnaire of alcoholic craving (ECCA Questionnaire: Behavior and Cognition in Relation to Alcohol: French translation and validation of the Obsessive-Compulsive Drinking Scale]. AB - Clinical, neurobiological and neuropsychological hypotheses suggest that the dimension of alcohol craving includes the concept of both obsessive thoughts about alcohol use and compulsive behaviors toward drinking. Anton et al. (1995) developed a 14 items self-rating scale, the Obsessive Compulsive Drinking Scale (OCDS) which includes items for assessing three dimensions: global, and the obsessive and the compulsive subdimensions. In this study, we included 156 patients, 105 men and 51 women, who met DSM IV diagnostic criteria for alcohol dependence. The mean age of our population was 39.1 +/- 11.2 years without difference between sexes. We did not found any correlation between the CAGE score and the OCDS total score or the obsessive and compulsive subscores (respectively, r = .15, r = .10 et r = .18). Moreover, we did not found any correlation between OCDS scores and mean daily alcohol consumption (r = .18, r = .16, r = .19). This could indicate that the dimension measured by the scale was somewhat independent of actual drinking. As such, it might act as an independent measure of the "state of illness" for alcohol-dependent patients. The test-retest correlation for the OCDS total score was .95 and the obsessive and compulsive subscales test-retest correlations were .93 and .89 respectively. The internal consistency of the items of the OCDS was high (alpha = .89). Principal component analysis had identified in the french version of the OCDS, three factors accounting for 63.5% of the total variance. These results indicate that the french version of the OCDS seems to validly measure a dimension of alcohol dependence. The ease of administration, reliability, and concurrent validity of the OCDS makes it particularly useful as an outcome measurement tool for various clinical therapeutic protocols in alcoholism. PMID- 9850817 TI - [Psychometric evaluation of the Hare Psychopathy Questionnaire in a Belgian prison population]. AB - The PCL-R (Hare, 1991) presents high reliability and validity characteristics. The instrument is the measure of choice for the assessment of psychopathic personality. The PCL-R has never been submitted to psychometric analysis in french speaking european samples. The study examines the psychometric characteristics of the PCL-R on an Belgian prison sample. The study assessed the inter-raters reliability parameters, the Alpha Cronbach coefficient and the PCL-R mean scores. The two correlated factors extracted were compared with those of others European and Nord American samples. The study also described the concomitant validity of the instrument by computing the correlations between the PCL-R scores and the type of offence. PMID- 9850818 TI - [Vulnerability to schizophrenia. I: Familial nature of of neuropsychologic indicators]. AB - The existence of a genetic background is now a well admitted notion in schizophrenia, but some individuals at genetic risk for that disease could never manifest it at a clinical level. However, several vulnerability models could help us to identify such individuals. According to them, when similar perturbations at a given test are observed both in clinically stable schizophrenics and their nonschizophrenic first degree relatives, this test could be qualify as an indicator of the vulnerability to schizophrenia. In literature, that seems the case for several neuropsychological tasks, exploring attentional abilities (degraded version of the Continuous Performance Task [DS-CPT], and Span Of Apprehension task [SOA]) and executive functions (Wisconsin Card Sorting Test [WCST]). Our study was undertaken to replicate literature data and to further explore the relationship between these three neuropsychological markers. For that purpose, performances at DS-CPT, SOA and WCST were assessed among 18 clinically stable schizophrenics, 18 of their biological full siblings and 15 unrelated control subjects matched with the two others groups for several socio-demographic factors. Comparisons were performed by non parametric analysis (Kruskal-Wallis one way ANOVA, and Mann-Whitney). Compared to controls, the siblings group performances were significantly impaired on the three tasks, while they did not statistically differ from the schizophrenic ones. No relationship was observed between the markers, except for the "d'" index at DS-CPT and the number of successfully performed categories at the WCST. Results from the sibling group suggested that the observed impaired neuropsychological performances may actually represent indicators of the genetic vulnerability to schizophrenia. Moreover, the generally admitted relationship between WCST poor performances and an impairment of the prefrontal cortex, lead us to hypothesize some role of this brain area in schizophrenia vulnerability. PMID- 9850819 TI - [Psychopharmacologic properties of Lippia multiflora]. AB - Lippia multiflora (L.m.) is a verbenacea used in Congo as conventional tea decoction. No traditional indication is known in this country. Nevertheless, in Ghana the plant is used for the treatment of arterial hypertension. The aim of this study is to investigate the psychotropic activity of the aqueous extract of L.m. using the classical tests of experimental psychopharmacology. The extract of L.m. is constituted by lyophilisated powder obtained from an infusion of dried leaves. Different doses are prepared: 200, 400, 600, 800, 1,000 and 1,200 mg/kg dissolved in 1 ml of NaCl 0.9%. L.m. is administered by intraperitoneal or oral route. The wistar rats of both sexes, weighing between 150-200 g, are used. Animal's behaviour is observed macroscopically. The spontaneous motor activity is appreciated by using the number of squares crossed by animal with the four paws in ten minutes (Martin and al. method slightly modified). The rectal temperature is measured. The effect of L.m. on stereotypies induced by apomorphin and anesthesia induced by phenobarbital are studied. The traction test is used to investigate the muscle relaxant effect of L.m. and analgesic activity is evaluated by using acetic acid and hot plate methods by comparison with diazepam 2 and 4 mg/kg. Fischer-t test is used for the statistical analysis of results. L.m. is well tolerated by rats. No mortality is observed with the doses used. So the doses of 200, 400 and 600 mg/kg were selected for experiments. At theses doses L.m. caused: a precocious ataxia, a sedation, a ptosis and a yellow coloration of urines, these effects are dose dependent; a significant reduction of spontaneous motor activity: control 61.60 +/- 6.48, L.m. 200: 16.40 +/- 5.68 (P < 0.01), L.m. 400: 12.20 +/- 2.01 and L.m. 600: 9.60 +/- 1.90 (P < 0.01); no modification of rectal temperature and apomorphin stereotypies; a reduction of sleep latence: control 22.40 +/- 1.89 min, L.m. 200: 17.20 +/- 2.74 min (P < 0.01), L.m. 400: 13.80 +/- 1.81 min (P < 0.01) and L.m. 600: 13.40 +/- 2.16 min (P < 0.01); a potentiation of phenobarbital anesthesia: L.m. 200: 209.80 +/- 29.58 min (N.S.), L.m. 400: 336.40 +/- 22.23 min (P < 0.01), L.m. 600: 342.20 +/- 16.28 min (P < 0.01) and control: 199.40 +/- 2.90 min; an increase at the dose of 400 mg/kg of the time necessary for the restoration of the paws to the metallic bar in the traction test: control; 0.8 +/- 0.1 s, L.m. 400: 7.04 +/- 2.29 s (P < 0.05); a reduction of abdominal cramps induced by acetic acid. This number is respectively 18.40 +/- 4.49 (P < 0.05); 15.00 +/- 2.90 (P < 0.01), 14.20 +/- 3.89 (P < 0.01), 11.60 +/- 4.75 (P < 0.01), 13.00 +/- 2.00 (P < 0.01) and 33.80 +/- 5.04 for L.m. 200 mg/kg, L.m. 400 mg/kg, L.m. 600 mg/kg, Diazepam 2 and 4 mg/kg and control; an increase of reaction time on the hot plate: L.m. 200: 3.26 +/- 0.46 s (N.S.), L.m. 400: 4.50 +/- 0.80 s (P < 0.01), L.m. 600: 10.50 +/- 1.56 s (P < 0.001), diazepam 2 mg/kg: 2.90 +/- 0.51 s (N.S.), diazepam 4 mg/kg: 5.90 +/- 1.09 s (P < 0.01) and control 2.10 +/- 0.26 s. Those results demonstrated that L.m. possess a tranquilizer and analgesic activities as Diazepam. But, anticonvulsant and anxiolytic tests are necessary to confirm the psychopharmacological profile of this medicinal plant. PMID- 9850820 TI - [Effects of antidepressants on symptoms, of inhibition]. PMID- 9850821 TI - [Value of tiapride for agitation in the elderly. Review of published studies]. AB - Among elderly disruptive behavior agitation and aggressiveness are frequent and often related to dementia. They are known for increasing the risk of patients' institutionalization and causing distress to families and care givers. When a drug prescription is required, physicians have to take into account the high sensitivity of elderly. Tiapride is an atypical neuroleptic which acts preferentially on D2 and D3 dopaminergic receptors. Several papers concerning results of clinical trials conducted in Europe and Japan have been published and reviewed in this article. The interest of tiapride in the treatment of elderly agitation and aggressiveness has been assessed in four double blind clinical trials including more than 700 patients. The efficacy of tiapride (aggressiveness, agitation, delusion and wandering) was demonstrated in a trial versus placebo (p = 0.027) including 324 patients treated for 28 days with 75 to 150 mg/d. Furthermore the superiority of tiapride (175 to 450 mg/d) on chlorpromazine (18 to 112.5 mg/d) was shown in two trials where 262 patients were treated for four weeks, and a recent survey highlighted that tiapride (400 mg/d) is as efficient as melperone (100 mg/d), the only neuroleptic to be indicated in treatment of elderly agitation and aggressiveness in Germany. Besides, 30 open clinical trials including around 1,000 patients have been conducted and have shown homogeneous and positive results. In the two trials versus chlorpromazine, the safety of tiapride was better, with especially less drowsiness, extrapyramidal symptoms, and dry mouth. Compared to lorazepam, in healthy subjects, tiapride caused less memory impairment. European and Japanese open studies confirmed the safety of tiapride in the elderly. In conclusion, tiapride at doses of 100 to 300 mg/d, appeared to be a therapy of elderly agitation and aggressiveness, at less as efficient as the other drugs used in this indication. Furthermore, the safety of tiapride is an advantage compared to benzodiazepines and neuroleptics. PMID- 9850822 TI - [alpha-beta L-aspartate magnesium in treatment of chronic benzodiazepine abuse: controlled and double-blind study versus placebo]. AB - OBJECTIVE: To evaluate the efficacy of alpha-beta L-Aspartate Magnesium (Asp Mg) in discontinuation of long-term benzodiazepine use and to search for a predictive model of success for BZD cessation. METHOD: Using a double-blind procedure, 144 patients selected as chronic users of one of 3 BZD lorazepam, alprazolam or bromazepam (duration of use > 6 months; regular dose > or = 3 mg lorazepam equivalent) and with clinical remission (score on Hamilton-Anxiety < 14; Raskin Depression < 6) had entered a controlled study (versus placebo) and were randomized in two parallel groups. The trial was conducted on 3 consecutive phases (co-administration of Asp Mg or placebo with BZD during 1 month; gradual taper of BZD during 1 month; follow-up during a third month after complete BZD discontinuation, with urinary BZD control on d75 and d90). RESULTS: The intent-to treat analysis showed at the endpoint an overall rate of 80% of "BZD discontinuation" and of 35.4% of "BZD cessation without withdrawal" in the total population (no significant intergroup differences were observed on these rates). However, there were some tendencies to positive differences between Asp Mg versus placebo on the following: 1) prolonged delay of BZD use if reintake (30 days vs 20 days, p [log-rank] = 0.5); 2) reduction of withdrawal intensity: 11% of important difficulties during BZD cessation versus 23% with placebo (p = 0.2) and on Benzodiazepine Withdrawal Symptoms Questionnaire (BWSQ) (final score 4.0 vs 4.8, p = 0.10); 3) lower modification of anxiety during BZD tapering and discontinuation (rate of increase on HAM-A between d30-d90 of 6% vs 23% in placebo group, p = 0.10). Moreover, 3 predictive factors of "success" (BZD cessation without withdrawal phenomenon) were identified by uni- and multivariate analysis with logistic regression: chronicity of anxiety disorder (p = 0.04) and amplitude of BWSQ change during tapering phase (p < 0.0001) as negative factors; and initial score of Speilberger Anxiety Inventory "Anxiety-Trait" (p = 0.002) as positive factor. A predictive model is constructed according to these 3 parameters. Further clinical trials are needed to explore the benefits of alpha beta L-Aspartate Magnesium in different criteria of prescription (dosage, duration of treatment, repetitive cures...). PMID- 9850823 TI - [Phototherapy: clinical and therapeutic evaluation of a 2-year experience]. AB - The concept of seasonal periodicity in psychiatry has been studied for thirty years in different domains, particularly in that affective disorders. It seems to be established that winter seasonal depressions are correlated to a particular vulnerability to the decrease in ambient light; this led directly to the development of phototherapy as primary treatment of SAD, simultaneously to the development of the clinical concept. Since the first demonstration by Rosenthal of the efficacy of phototherapy in SAD, numerous studies have confirmed that bright light provides rapid and effective treatment for this type of disorder. Phototherapy has been used at the Centre Psychotherapique des Hopitaux Civils de Colmar for five years in three ways: as an alternative treatment to medication, in addition to antidepressive medication, and as primary treatment of seasonal depression. We describe the evaluation protocol of this treatment, which was established in collaboration with the Psychiatric Clinic of the University of Basel. The principal inclusion criterion for this study was the presence or regular phases of depression or loss of energy in autumn and winter; the regularity of the appearance of symptoms in winter was predominant over the severity of the symptoms, their seasonal occurrence being the most important predictive factor in favour of phototherapy. Our prospective study enabled us to include, over a period of two years, 18 subjects presenting very different symptomatology. Though the sample was heterogeneous, the results of treatment efficacy are satisfactory: we note effectively a decrease of 67.5% in the modified Hamilton scale even though strict criteria for seasonality (Seasonal Screening Questionary, Rosenthal's Criteria) were not always respected. These data should lead us to widen the indication for phototherapy, particularly in patients presenting mood disorders which are not exclusively seasonal, and support the accumulating experimental data for the selective antidepressive effect of light. PMID- 9850824 TI - [A case of Parkinson syndrome secondary to combined amineptine and bromazepam abuse]. AB - A case report of parkinsonism secondary to chronic abuse of amineptine (3 gr/day) and bromazepam (35 mg/day) in a patient diagnosed of borderline personality disorder is presented. The patient did not take any other drugs and he was not recently on neuroleptic treatment; he recognized the abuse of amineptine, as a stimulant, and the bromazepam abuse, looking for a relief to the excessive anxiety secondary to amineptine. The parkinsonism improved after removing both drugs and taking biperiden and diazepam; lastly the patient was discharged without any medication. The patient did not suffer from other complications associated with amineptine or bromazepam abuse. There are some cases reported of parkinsonism secondary to benzodiazepines but there is none secondary to amineptine. We present a short review of the possible responsible mechanisms, thinking on a complex interaction of both drugs on dopamine and its modulatory systems. PMID- 9850825 TI - [Communication on the Internet. Possibilities and risks of the Internet for psychiatry]. AB - The article is preceded by an introduction to world-wide communication networks. Some important basic features are explained. The major impact is on temporary and future aspects of the Internet for psychiatry. A variety of advantages in World Wide Web telemedicine for patients, clients, psychotherapists, clinicians and scientists are extensively demonstrated. Useful and established possibilities are discussed. Finally, the risks of this kind of communication are shown and listed. PMID- 9850826 TI - [Sensation seeking--critical review of personality psychological concepts and its application]. AB - The term "sensation seeking" is part of a self-contained theory on personality psychology. It is meant to denote a disposition--peculiar to a certain personality, probably genetically founded and correlating with biological, f. e. neuroendocrine measures--to explore one's social environment in order to find new and diversified stimuli. In their work, researchers mainly refer to Zuckerman's Sensation Seeking Scale in its fifth version (SSS-V), which distinguishes between four aspects of sensation seeking based on factor analysis of the 40 SSS-V-items. There is partly an overlapping of the phenomenon with impulsiveness and extraversion. Whereas application studies show inter alia links between sensation seeking and dangerous driving or dangerous sexual practices, data on the expression of this trait in addictive persons do not point to definitely consistent links. In forensic terms, there is on a link between "boredom susceptibility"--the inability to stand uneventful, monotonous situations--and "disinhibition"--a lack of control of impulses--figuring as subscales of the SSS V, on the one hand, and hyperkinetic attention deficit syndromes and "disactualisation weakness", as described by Janzarik, on the other. At present, there is no German-language SSS-inventory in line with basal testing theory requirements. Because of the apparently different modi of sensation seeking influenced by other cultural traditions, such a scale must be basically adapted to the life-style in Western Europe. PMID- 9850827 TI - [The costs of schizophrenia]. AB - Schizophrenia is regarded as the most expensive mental illness because of its specific symptoms and characteristics (e.g. early onset, often chronic course, high rates of readmission to hospital treatment, high rate of disabilities and extensive rehabilitative interventions), which prove to be extremely costly. Despite this, studies on the financial aspects of schizophrenia or the provision of care for schizophrenic patients has become an issue of psychiatric research only since the beginning of the reform of mental healthcare. Early cost studies had been conducted in the United States (US) during the fifties. Since then, they have grown in number not only in the US and in Great Britain, but also on the Continent of Europe. On an international level, comprehensive literature concerning methodology has attempted to establish cost studies as an integral part of mental health services research. Germany, however, is far behind international developments. Although the fundamental lack of empirical data on costs both in psychiatry as a whole and in schizophrenia had already been ascertained in a large national survey called "Psychiatrie-Enquete" in the mid seventies, little has changed since then. One reason for these possibly great methodological problems is associated with the assessment of cost data in fragmented community mental health care networks, which in Germany include the additional obstacle--unlike abroad--of non-availability of access to data from case registers. Psychiatric case registers are not permitted in Germany because of very strict data protection laws. Despite the problems in methodology, there is an urgent need in Germany to remedy the lack of cost data for schizophrenic patient care. The pressure of curbing costs in health care will probably force the German mental health care services to provide detailed cost data with regular reports in the future. PMID- 9850829 TI - [Psychopathologic symptoms and cognitive test performance in schizophrenic patients]. AB - Psychopathological symptoms and cognitive test performance were examined in 34 acute schizophrenic patients. The results of a clusteranalytic approach in order to distinguish groups of patients with different syndromes were disappointing. Three dimensions of negative, hallucinatory-delusional, and disorganised symptoms could be established by factor analysis. The disorganised symptom dimension showed strong and significant relations to mnestic and intellectual impairments of the patients. Hallucinatory-delusional symptoms were related to deficits in tests of visual memory and visual search. Negative symptoms were not related to cognitive impairments of the patients. The results are discussed in respect of other studies reporting correlations of schizophrenic symptoms and cognitive disturbances, and with regard to hypotheses of brain dysfunction in schizophrenia. In future research, consideration of the three main dimensions of schizophrenic symptoms could be useful to reduce the heterogeneity of schizophrenic samples. PMID- 9850828 TI - [Violent and non-violent methods of parasuicide: what determines the choice?]. AB - Patients with violent methods of parasuicide share a number of common characteristics with those who complete suicide. They can be differentiated from patients with non-violent methods. Whereas surgery is usually the first-line care in cases of violent parasuicide, detoxification and/or psychiatric hospitalisation are first employed with non-violent parasuicide. Therefore it is important to know the specific needs and characteristics of both patient groups, as it may be hypothesised that patients with violent methods are at specifically high risk of committing suicide. As part of the WHO/EURO Multicentre Study on Parasuicide, we examined 120 cases of parasuicide in hospitals of the Bonn area with the instrument EPSIS 1. Violent and non-violent methods were differentiated following the WHO X-classification. RESULTS: The following independent variables differentiate between violent and non-violent methods and predict the choice of violent methods: Diagnosis (schizophrenia vs. other diagnoses, p = 0.00027), gender (male, p = 0.04), high score of anger as a trait in State-Trait-Anger Scale (p = 0.017), poor mental health within the last 3 months (p = 0.058), time of parasuicide after 6 p.m. (p = 0.024). A higher number of previous parasuicides (p = 0.008) and unemployment (p = 0.047) were predictive of the choice of non violent methods. A logistic regression analysis generated a model including the independent variables diagnosis, gender and "anger". Suicidal intent, sociodemographic variables, motives of parasuicides and "life events" did not discriminate between violent and non-violent methods. PMID- 9850830 TI - [Dissociation and schizophrenia. Schizophrenias--a dissociative nosopoietic construct?]. AB - The historical roots of the nosopoietic construct schizophrenia are traced: the nosographic construction by Kraepelin, an act of unification and split at the same time. The name schizophrenia reflects the tradition of the model of dissociation which is aetiologically connected with the idea of weakness of the synthetic power of the psyche: psychasthenia. In the establishment of the supposed nosological entity schizophrenia, the name-giving idea of dissociation was almost forgotten. Thus, nowadays Dissociative Disorders are classified in ICD and DSM separately from schizophrenic disorders, but it must be borne in mind that the schizophrenic disorders represent the most severe of disorders, hypothetically based on dissociation, namely, fragmentation of the ego. PMID- 9850831 TI - [Frequency and comorbidity of social anxiety and social phobia in adolescents. Results of a Bremen adolescent study]. AB - Using data from the Bremen Adolescent Study, this report presents findings on the frequency, comorbidity and psychosocial impairment of social phobia and social fears among 1035 German adolescents of 12-17 years of age. The adolescents were randomly selected from 36 schools in the city and provincial government area of Bremen, Germany. Social phobia and other psychiatric disorders were coded based on DSM-IV criteria using the computerized personal interview of the Munich version of the Composite International Diagnostic Interview. Seventeen (1.6%) of the adolescents had met the DSM-IV criteria for social phobia at some time in their life. More girls than boys were diagnosed as suffering from social phobia. The incidence of the disorder increased with age. The lifetime frequency of social fears is much higher. The most common types of feared social situations were fear of doing something in front of other people, followed by public speaking. Social phobia very often co-occurred with depressive disorders, somatoform disorders and disorders caused by excessive or inappropriate consumption of substances. Over 94% of those with social phobia and 54.4% with any social fears were severely impaired in their daily life during the worst episode. Despite the high level of psychosocial impairment, only a small portion of the cases received professional help. PMID- 9850832 TI - [Clinical studies on renal trauma]. AB - A total of 18 kidneys in 17 patients with renal trauma were treated at the Department of Urology, Mitoyo General Hospital between April 1991 and August 1997. Patients were between 10 and 88 years old (median; 52). Eleven of them were male and 6 were female. The main cause of injury was a traffic accident in 11 cases (64.7%), which was similar in tendency to previous reports in this country. According to the Classification of Renal Injury by the Japanese Association for the Surgery of Trauma in Japan, there were 8 cases (47.1%) of Type I (sub capsular injury) and Type II (superficial injury) that was classified a minor injury, and there were 9 cases (52.9%) of Type III (deep injury), Type IV (pedicle injury) and open injury that was classified as a major injury. Surgical treatment was performed in 8 cases (47.1%), which were all classified as a major injury. Posttraumatic plasma lactate dehydrogenase (LDH) was elevated immediately in case of major injury in comparison with a case of minor injury. Serum LDH may be useful as a parameter of the degree of renal trauma. PMID- 9850833 TI - [Clinical studies on the prognostic factors in prostate cancer]. AB - To evaluate the prognostic factors and the outcome of treatment, a retrospective study was done on 141 patients with prostate cancer who were newly diagnosed at Kitano Hospital between January 1985 and November 1996. In recent years, the number of patients and the ratio of low stage cancer have increased. The overall 5-year crude survival rate was 49.9%. The 5-year crude survival rate for clinical stage A, B, C and D was 67%, 70%, 62% and 30% respectively. The overall 5-year disease-specific survival rate was 65.6%. The 5-year disease-specific survival rate for clinical stage A, B, C and D was 100%, 89%, 72% and 42%, respectively. By univariate analysis, clinical stage, Gleason score, prostate specific antigen (PSA) level, and patient age were prognostic factors for disease-specific survival of prostate cancer. According to Cox's regression analysis by the stepwise forward regression method, clinical stage and Gleason score were selected as more valuable prognostic factors than PSA level, patient age, comorbidity, and initial treatment. In Gleason score 2 to 8, the prognosis became significantly worse as clinical stage advanced, but in Gleason score 9 and 10 the prognosis was poor regardless of clinical stage. PMID- 9850834 TI - [Clinical experience of transurethral collagen injections for urinary stress incontinence--analysis of subjective symptoms]. AB - We evaluated the results of transurethral collagen injections for urinary stress incontinence. Twenty five women (mean age was 61.3 years) with urinary incontinence were treated with transurethral collagen injections using local or spinal anesthesia. The mean follow-up was 11.7 months (range 2 to 30). We examined the results based on subjective symptoms for incontinence. We could judge convalescence efficacy to some degree 1 month after operation, but patient age, type of stress incontinence, pad test and volume of collagen were not significantly different between patients who were cured and those not cured. Of the patients who needed injections more then 2 times, treatment was effective in type III patients. The patients whose symptoms were improved 3 months after operation wanted a re-operation when their incontinence recurred. Injection of transurethral collagen appears to be a safe and effective method for treating urinary incontinence. This procedure is a first choice for urinary incontinence. PMID- 9850835 TI - [A case of accessory spleen presenting as retroperitoneal tumor]. AB - We report the 13th case of accessory spleen in the retroperitoneum in the world literature. A 53-year-old man visited our hospital complaining of lassitude. Laboratory studies revealed significant anemia. Several imaging studies revealed a large retroperitoneal tumor (70 x 63 x 48 mm) in the medial aspect of the lower pole of the left kidney. The tumor was supplied arterially from the first and second lumbar arteries. Since the tumor adhered tightly to the left kidney, the kidney was resected as well as the tumor. The tumor was very hemorrhagic. On histological examination, the tumor exhibited a structure typical of splenic tissue. The retroperitoneal accessory spleen is very rare, and the literature on it is reviewed here. PMID- 9850836 TI - [A case of surgical resection for metachronous multiple liver metastases of chromophobe cell renal carcinoma]. AB - A 64-year-old female was hospitalized because of multiple liver metastasis, during the postoperative observation of chromophobe cell renal carcinoma. Injection of Epirubicin/Lipodol into the right hepatic artery followed by transcatheter arterial embolization was performed two times, and the metastatic foci responded favorably to this therapy. Since no new metastatic tumors were observed in the liver nor any other organs, a partial hepatectomy was thus performed. The liver tumor was pathologically diagnosed to be metastasis of the chromophobe cell renal carcinoma. The postoperative course has been uneventful and at present, 6 months after the operation, no further tumors have been observed. PMID- 9850837 TI - [Urolithiasis associated with Crohn's disease: a case report]. AB - We report a case of urolithiasis caused by surgical treatment for Crohn's disease. A 28-year-old woman was referred to our department for further examination of renal stones from the medical department in September, 1995. She suffered from Crohn's disease and had a history of jejuno-ileal resection because of perforation of the ileum in 1988. Radiographs revealed multiple bilateral renal stones, and the urine oxalate concentration was elevated. She was treated with extracorporeal shock wave lithotripsy and the administration of sodium bicarbonate and citrate, but these treatments did not prevent recurrence and enlargement of stones. Renal function was gradually worsened and we performed transurethral lithotomy and percutaneous nephrolithotripsy. The stones were mainly composed of oxalate calcium monohydrate. A renal biopsy was performed at the operation, showing deposition of crystals in almost all renal tubules. Diet therapy (low oxalate and low fat) and the administration of sodium bicarbonate and citrate were performed strictly and recurrence was not recognized 10 months after complete removal of the stones. PMID- 9850838 TI - [2,8-dihydroxyadenine urolithiasis due to partial deficiency of adenine phosphoribosyltransferase: a case report]. AB - A 35-year-old female was referred to our clinic with a complaint of left flank pain in 1993. Drip infusion pyelography showed a filling defect of 25 x 24 mm in size in the left ureteropelvic junction. Computed tomography and ultrasonography revealed it as the renal stone. Percutaneous nephroureterolithotomy and extracorporeal shock-were lithotomy were performed. The stone was composed of 2,8 dihydroxyadenine (DHA). The patient was diagnosed as having a partial deficiency of adenine phosphoribosyltransferase (APRT) from the low APRT activity and a genotype of a compound heterozygote APRT*J/APRT*Q0 by T-cell analysis. The urinary excretion of 2,8-DHA crystals disappeared by the postoperative treatment with allopurinol. Cases of 2,8-DHA urolithiasis reported in the Japanese literature are discussed. PMID- 9850839 TI - [A case of xanthogranulomatous pyelonephritis followed by computed tomographic scan]. AB - Since it is difficult to differentiate xanthoglanulomatous pyelonephritis (XGP) from renal cancer, most cases of XGP are diagnosed after nephrectomy. We report a rare case of XGP followed by abdominal computed tomographic (CT) scan. A 65-year old woman was admitted with high fever. Drip infusion pyelography showed a right renal stone but did not visualize the right kidney. A swollen right kidney was revealed on abdominal CT scan. Diabetes mellitus was detected by blood examinations and XGP was suspected. Conservative therapy with antibiotics was performed, the symptoms disappeared within 2 weeks and the right atrophic kidney was revealed on abdominal CT scan 12 months after onset. Histopathological findings on a renal biopsy specimen obtained 1.5 months after treatment were compatible with XGP. The diagnosis of XGP will become easier by using a combined approach including radiographic findings, physical finding and other data. PMID- 9850840 TI - [A case of renal pelvic carcinoma in a completely duplicated pelvis and ureter]. AB - We report a case of renal pelvic carcinoma with a completely duplicated pelvis and ureter. A 72-year-old man was referred to our hospital because of asymptomatic and macroscopic hematuria. Excretory urography and retrograde pyelography revealed a tumor in the left upper renal unit of a completely duplicated pelvis and ureter. Left total nephroureterectomy was performed. Histological diagnosis was transitional cell carcinoma. This is the 8th case of upper urotherial tumor occurring in a completely duplicated pelvis and ureter in the Japanese literature. PMID- 9850841 TI - [A case of ureteral polyp prolapsing into bladder in a 3-year-old boy]. AB - A 3-year-old boy was referred to us with a diagnosis of bladder tumor. On cystoscopy, a yellowish-white pedunculated tumor was found at the right trigone and biopsy revealed an inflammatory change. Computed tomographic (CT) scan showed a soft tissue density from the lower ureter to the bladder. Open surgery confirmed a ureteral polyp originating from the lower ureter prolapsing into the bladder, which mimicked bladder tumor. The need of open surgery in such cases is discussed. PMID- 9850842 TI - [Leiomyosarcoma of the prostate: report of two cases]. AB - We herein present two cases prostatic leiomyosarcoma. The first case was in a 45 year-old man who presented at our department with the chief complaints of pain on voiding and pollakisuria on November 13, 1996. Ultrasonography and computed tomographic (CT) scan revealed a prostatic tumor. A histological examination of biopsy specimens revealed leiomyosarcoma of the prostate. Total prostatectomy and partial cystectomy were performed. No adjuvant therapy was performed. He is still alive without disease 12 months after operation. The second case was in a 63-year old man who was admitted to our hospital for treatment of a lung tumor and colon polyp on February 28, 1997. CT scans showed a large prostatic tumor and multiple tumors in the lung, liver and bilateral kidneys. He was referred to our department for evaluation of the prostatic tumor. A transrectal needle biopsy of the prostate for histological diagnosis revealed leiomyosarcoma. No treatment was performed and he died 3 months later. In addition, 57 cases of prostatic leiomyosarcoma collected from the Japanese literature are also reviewed. PMID- 9850844 TI - [MRI findings in a testicular epidermoid cyst: a case report]. AB - A 21-year-old man was admitted with a painless mass in the left testis. Physical examination revealed a firm, small-finger sized, mass lesion with a smooth surface in the left testis. The ultrasonographic appearance was a hypoechoic lesion with an echogenic rim in the left testis. Magnetic resonance imaging (MRI) findings showed the echogenic rim to be a low-signal zone on both T1 and T2 weighted images. The center, on T2-weighted images, had a low signal intensity, with peripheral high signal intensity. Under a preoperative diagnosis of epidermoid cyst, the mass was resected surgically with preservation of the testis. Histopathological diagnosis confirmed an epidermoid cyst. Preoperative ultrasonographic and MRI findings suggested a benign lesion, thus permitting preservation of the testis. PMID- 9850843 TI - [A case of neutropenic enterocolitis in high dose chemotherapy with peripheral blood stem cell transplantation for relapsed testicular tumor]. AB - High dose chemotherapy with peripheral blood stem cell transplantation (PBSCT) is safe and useful for the treatment of refractory or relapsed testicular tumors, but is sometimes accompanied by serious complications. We report a case of relapsed testicular tumor complicated by neutropenic enterocolitis during high dose chemotherapy with PBSCT. PMID- 9850845 TI - [Cavernous hemangioma of the scrotum: a case report]. AB - A 19-year-old male consulted our department, complaining of a painful scrotal mass. On palpation, the mass was discriminated from testis, epididymis, and spermatic cord in the left scrotum. He underwent total excision of the mass. Pathologic examination revealed cavernous hemangioma. Scrotal hemangioma is a relatively rare lesion and this is the 37th case reported in Japan. PMID- 9850846 TI - [Clinical utility of the free prostate specific antigen (PSA), alpha 1 antichymotrypsin-complexed PSA, and free/total PSA ratio using the specific and sensitive enzyme-linked immunosorbent assay "E-plate EIKEN PSA"]. AB - We studied the clinical significance of serum prostate specific antigen (PSA) ratio: free-PSA/total-PSA and free-PSA/complex-PSA to discriminate between prostate cancer (PC) and prostate benign disease (non-PCa) by using total-PSA, alpha 1-antichymotrypsin complexed (complex)-PSA and free-PSA enzyme-linked immunosorbent assay (ELISA) kits newly developed at EIKEN Chemical Co, Ltd. Fre PSA and complex-PSA ELISA kits demonstrated high sensitivity and specificity. Total-PSA ELISA kit also demonstrated equimolarity for free-PSA and complex-PSA. On the total-PSA range of 4-10 ng/ml, free-PSA/total-PSA% (f/t%) and free PSA/complex-PSA% (f/c%) were very useful to discriminate between PCa and non-PCa by receiver operating characteristic curve analysis as well as PSA density (PSA D) but not free-PSA level. F/t% and f/c% were even useful to discriminate early stage PCa (i.e. A1 or B0) from non-PCa by the Mann-Whitney U-test. PMID- 9850847 TI - Cell fate in the immune system: decisions, decisions, decisions. PMID- 9850848 TI - Role of interleukin-7 in T-cell development from hematopoietic stem cells. AB - All lymphocytes are derived from hematopoietic stem cells (HSC). The interleukin 7 receptor (IL-7R) transduces non-redundant signals for both T and B-cell development from HSC. The upregulation of the IL-7R occurs at the stage of the clonogenic common lymphoid progenitor, a recently identified population that can give rise to all lymphoid lineages (T, B and natural killer cells) at a single cell level. The IL-7R plays a critical role in the rearrangement of immunoglobulin heavy chain genes required for B-cell development. IL-7R expression is critically regulated in developing thymocytes; thymocytes that fail the positive selection process downregulate the IL-7R, but those undergoing positive selection upregulate or maintain IL-7R expression. Recent data indicate that IL-7 signaling enhances the survival of developing thymocytes and mature T cells, presumably by its upregulating Bcl-2. Detailed analysis of the signaling cascades activated by the IL-7R may help to reveal the differential roles of IL-7 signaling in T and B-cell development. PMID- 9850849 TI - Natural killer and T cells of innate and adaptive immunity: lymphoid compartments with different requirements for common gamma chain-dependent cytokines. AB - A group of cytokines, including interleukin-2, -4, -7, -9 and -15, are related through the usage of a shared receptor subunit, the common cytokine receptor gamma chain, gamma c. gamma c-dependent cytokines critically affect the development and maintenance of the lymphoid system. This review will highlight our current knowledge on the gamma c-dependent cytokine network and on the non redundant roles that these cytokines play in the development and homeostasis of T and natural killer cells involved in innate and adaptive immunity. PMID- 9850850 TI - The linkage between T-cell and dendritic cell development in the mouse thymus. AB - Thymic dendritic cells (DC) mediate negative selection at a relatively late stage of the T-cell developmental pathway. We present evidence that the development of thymic DC and of T-lineage cells is linked via a common precursor at an early stage of thymocyte development. T-lineage precursor populations from the adult mouse thymus, prior to T-cell receptor gene rearrangement, display a capacity to produce DC as well as T cells in the thymus, and are very efficient precursors of DC in culture. These lymphoid/DC precursors have little capacity to form myeloid cells, indicating that thymic DC are a lymphoid-related rather than myeloid related lineage. In contrast to myeloid-related DC, granulocyte-macrophage colony stimulating factor is not required for the development of these lymphoid-related DC in vivo or in vitro. DC can develop in mutant mice lacking mature T cells, provided the common precursors are present. However, in mutant mice lacking functional Ikaros transcription factors, there are deficiencies in lymphoid precursor cells, in mature lymphoid cells and in DC. PMID- 9850851 TI - Natural killer cell differentiation: insights from knockout and transgenic mouse models and in vitro systems. AB - In the last few years, the routine development of knockout and transgenic mice and the ease with which rare progenitor populations can be isolated from hematopoietic organs and cultured in vitro has facilitated significant advances in understanding the lineage and development of natural killer (NK) cells. Fluorescence-activated cell sorter analyses have identified a common lymphoid progenitor capable of giving rise to NK, T, and B cells, confirming the lymphoid origin of NK cells. Knockout and transgenic mouse models have pointed to an absolutely critical role for signals sent through the interleukin (IL)-2/15 receptor beta (CD122) chain and common gamma (gamma c) chain for NK development. Such signals are likely relayed inside the cell by the tyrosine kinase Jak3, which associates with gamma c. Recently developed IL-15 and IL-15 receptor alpha knockout mice have pinpointed IL-15 as the mediator of this signal. Other mouse models have indicated an unexpected role for flt3 ligand in early NK-cell development as well as minor roles for stem cell factor and IL-7 in expanding NK cell progenitor numbers. Finally, in vitro culture systems have proven useful in identifying the point in NK development at which each of these signals is critical. PMID- 9850852 TI - Lineage commitment and differentiation of T and natural killer lymphocytes in the fetal mouse. AB - T cells and natural killer (NK) cells are presumed to share a common intrathymic precursor. The development of conventional alpha beta T lymphocytes begins within the early fetal thymus, after the colonization of multipotent CD117+ precursors. Irrevocable commitment to the T lineage is marked by thymus-induced expression of CD25. However, the contribution of the fetal thymus to NK lineage commitment and differentiation remains largely unappreciated. Recently, we demonstrated that the development of functional mouse NK cells occurs first in the fetal thymus. Moreover, the appearance of mature fetal thymic NK cells (NK1.1+/CD117-) is preceded by a thymus-induced developmental stage (NK1.1+/CD117+) that marks lineage commitment of multipotent hematopoietic precursors to the T and NK-cell fates. Commitment to the T/NK bipotent stage is induced by fetal thymic stroma, but is not thymus dependent. Recent data indicate that CD90+/CD117lo fetal blood prothymocytes exhibit NK lineage potential and are phenotypically and functionally identical to fetal thymic NK1.1+/CD117+ progenitors. This finding also indicates that full commitment of circulating precursors to the T-cell lineage occurs after thymus colonization. In this review, we discuss recent insights into the cellular and molecular events involved in fetal mouse T and NK lineage commitment and differentiation to unipotent progenitors. PMID- 9850853 TI - Early stages in the development of human T, natural killer and thymic dendritic cells. AB - T-cell development is initiated when CD34+ pluripotent stem cells or their immediate progeny leave the bone marrow to migrate to the thymus. Upon arrival in the thymus the stem cell progeny is not yet committed to the T-cell lineage as it has the capability to develop into T, natural killer (NK) and dendritic cells (DC). Primitive hematopoietic progenitor cells in the human thymus express CD34 and lack CD1a. When these progenitor cells develop into T cells they traverse a number of checkpoints. One early checkpoint is the induction of T-cell commitment, which correlates with appearance of CD1a and involves the loss of capacity to develop into NK cells and DC and the initiation of T-cell receptor (TCR) gene rearrangements. Basic helix-loop-helix transcription factors play a role in induction of T-cell commitment. CD1a+CD34+ cells develop into CD4+CD8 alpha+ beta+ cells by upregulating first CD4, followed by CD8 alpha and then CD8 beta. Selection for productive TCR beta gene rearrangements (beta selection) likely occurs in the CD4+CD8 alpha+ beta- and CD4+CD8 alpha+ beta+ populations. Although the T and NK-cell lineages are closely related to each other, NK cells can develop independently of the thymus. The fetal thymus is most likely one site of NK-cell development. PMID- 9850854 TI - The role of the T-cell receptor (TCR) in alpha beta/gamma delta lineage commitment: clues from intracellular TCR staining. AB - T cells belong to two mutually exclusive lineages expressing either alpha beta or gamma delta T-cell receptors (TCR). Although alpha beta and gamma delta cells are known to share a common precursor the role of TCR rearrangement and specificity in the lineage commitment process is controversial. Instructive lineage commitment models endow the alpha beta or gamma delta TCR with a deterministic role in lineage choice, whereas separate lineage models invoke TCR-independent lineage commitment followed by TCR-dependent selection and maturation of alpha beta and gamma delta cells. Here we review the published data pertaining to the role of the TCR in alpha beta/gamma delta lineage commitment and provide some additional information obtained from recent intracellular TCR staining studies. We conclude that a variant of the separate lineage model is best able to accommodate all of the available experimental results. PMID- 9850855 TI - T-cell fate. AB - Recent studies suggest that lineage commitment steps, which occur during T-cell differentiation, follow principles in common with fate specification in simple invertebrates. Here we review T-cell development from the perspective of developmental biology. We present models for alpha beta vs gamma delta and CD4 vs CD8 lineage commitment that are consistent with previously published and newly presented experiments. PMID- 9850856 TI - Crucial function of the pre-T-cell receptor (TCR) in TCR beta selection, TCR beta allelic exclusion and alpha beta versus gamma delta lineage commitment. AB - The analysis of T-cell receptor (TCR) beta selection, TCR beta allelic exclusion and TCR beta rearrangement in gamma delta T cells from normal and pre-TCR deficient mice has shown that the pre-TCR has a crucial role in T-lymphocyte development: The pre-TCR is by far the most effective receptor that generates large numbers of CD4+8+ T cells with productive TCR beta rearrangements. In the absence of the pre-TCR, TCR beta rearrangement proceeds in developing cells irrespective of whether they already contain a productive TCR beta gene. The pre TCR directs developing T cells to the alpha beta lineage because gamma delta T cells from pT alpha-/- mice proceed much further in TCR beta rearrangement than gamma delta T cells from wild-type mice. It is argued that the pre-TCR commits developing T cells to the alpha beta lineage by an instructive mechanism, which has largely replaced an evolutionarily more ancient mechanism that involves stochastic alpha beta lineage commitment. PMID- 9850857 TI - Accessibility control of variable region gene assembly during T-cell development. AB - T-cell development is a complex and ordered process that is regulated in part by the progressive assembly and expression of antigen receptor genes. T cells can be divided into two lineages based on expression of either an alpha beta or gamma delta T-cell antigen receptor (TCR). The genes that encode the TCR beta and gamma chains lie in distinct loci, whereas the genes that encode the TCR alpha and delta chains lie in a single locus (TCR alpha/delta locus). Assembly of TCR variable region genes is mediated by a site-specific recombination process that is common among all lymphocytes. Despite the common nature of this process, recombination of TCR genes is tightly regulated within the context of the developing T cell. TCR beta, gamma and delta variable region genes are assembled prior to TCR alpha variable region genes. Furthermore, assembly of TCR beta variable region genes is regulated within the context of allelic exclusion. The regulation of rearrangement and expression of genes within the TCR alpha/delta locus presents a complicated problem. TCR alpha and delta variable region genes are assembled at different stages of T-cell development, and fully assembled TCR alpha and delta variable region genes must be expressed in distinct lineages of T cells, alpha beta and gamma delta, respectively. We have developed several experimental approaches to assess the role of cis-acting elements in regulating recombination and expression of TCR genes. Here we describe these approaches and discuss our analyses of the regulation of accessibility of the TCR beta and TCR alpha/delta loci during T-cell development. PMID- 9850858 TI - Developmental regulation of V(D)J recombination at the TCR alpha/delta locus. AB - The T-cell receptor (TCR) alpha/delta locus includes a large number of V, D, J and C gene segments that are used to produce functional TCR delta and TCR alpha chains expressed by distinct subsets of T lymphocytes. V(D)J recombination events within the locus are regulated as a function of developmental stage and cell lineage during T-lymphocyte differentiation in the thymus. The process of V(D)J recombination is regulated by cis-acting elements that modulate the accessibility of chromosomal substrates to the recombinase. Here we evaluate how the assembly of transcription factor complexes onto enhancers, promoters and other regulatory elements within the TCR alpha/delta locus imparts developmental control to VDJ delta and VJ alpha rearrangement events. Furthermore, we develop the notion that within a complex locus such as the TCR alpha/delta locus, highly localized and region-specific control is likely to require an interplay between positive regulatory elements and blocking or boundary elements that restrict the influence of the positive elements to defined regions of the locus. PMID- 9850859 TI - Thymocyte selection in Vav and IRF-1 gene-deficient mice. AB - T cells undergo a defined program of phenotypic and genetic changes during differentiation within the thymus. These changes define commitment of T-cell receptor (TCR) gamma delta and TCR alpha beta cells and lineage differentiation into CD4+ T helper and CD8+ cytotoxic T cells. T-cell differentiation and selection in the thymus constitute a tightly co-ordinated multistep journey through a network that can be envisaged as a three-dimensional informational highway made up of stromal cells and extracellular matrix molecules. This intrathymic journey is controlled by information exchange, with thymocytes depending on two-way cellular interactions with thymic stromal cells in order to receive essential signals for maturation and selection. Genetic inactivation of surface receptors, signal transduction molecules, and transcription factors using homologous recombination has provided novel insight into the signaling cascades that relay surface receptor engagement to gene transcription and subsequent progression of the developmental program. In this review we discuss molecular mechanisms of T lymphocyte development in mice that harbour genetic mutations in the guanine nucleotide exchange factor Vav and the interferon regulatory transcription factor 1 (IRF-1). We also propose a novel model of T-cell selection based on TCR alpha chain-directed signals for allelic exclusion and TCR alpha based selection for single receptor usage. PMID- 9850860 TI - The Syk family of protein tyrosine kinases in T-cell activation and development. AB - The processes of T-cell development and activation employ similar immature and mature receptors as well as similar signal transduction pathways to achieve different outcomes. Many signaling molecules are shared between the receptor signaling pathways, including two families of cytoplasmic protein tyrosine kinases, the Src family and the Syk family. The two Syk family members expressed in T cells, Syk and ZAP-70, are structurally similar but are expressed at different times during thymic development and during T-cell activation. These two kinases, although they share many physical features, differ in terms of biochemical activity and regulation. We discuss the overlapping and distinct characteristics of Syk and ZAP-70 in T-cell signaling and the potential biological importance of their differences. PMID- 9850861 TI - Molecular requirements for lineage commitment in the thymus--antibody-mediated receptor engagements reveal a central role for lck in lineage decisions. AB - Recent experiments in our laboratory have focused on the receptor engagements required for the differentiation of fully mature, single positive thymocytes from their double positive precursors. We have used a novel approach which involves the ligation of surface receptors on immature thymocytes with genetically engineered F(ab1)2 reagents, which, unlike conventional antibodies, do not aggregate the CD3 complex to such an extent as to induce extensive deletion of these cells. The experimental data presented in this review indicate that differentiation of the two mature CD4 and CD8 lineages occurs in response to distinct intracellular signals induced by particular receptor engagements. The data suggest that the tyrosine kinase p56lck (lck) plays a crucial role in determining lineage choice, in that maturation of thymocytes into the CD4 lineage occurs upon recruitment of active lck to the T-cell receptor (TCR)/CD3 complex, whereas CD8 maturation can be induced by CD3 ligation in the absence of co receptor-mediated lck recruitment. A central role for lck activity in determining the threshold for differentiation of the CD4 lineage is revealed in experiments with thymi deficient for a regulator of lck activity, CD45. A model of thymocyte differentiation is presented in which we propose that the relative balance of signals delivered by TCR engagement and lck activation determines lineage choice. PMID- 9850862 TI - CD4/CD8 lineage commitment: matching fate with competence. AB - The outcome of positive selection of T lymphocytes is that there is a close match between the lineage adopted by a particular cell (CD4+ or CD8+) and the specificity of the T-cell receptor for the class of Major Histocompatibility Complex molecule recognized. How this match is obtained has been a matter of debate. We review the evidence, from recent and older experiments, that indicates that the process follows a selective logic, rather than an instructive one. PMID- 9850863 TI - Thymocyte selection: not by TCR alone. AB - During development of T cells in the thymus, T-cell receptor (TCR)-mediated recognition of self-MHC/self-peptide complexes on thymic stroma dictates the developmental fate of immature CD4+CD8+ (double positive) thymocytes. Intriguingly, TCR-generated intracellular signals can elicit two entirely different cellular responses in such thymocytes: apoptosis or further differentiation. The critical issue in understanding end-stage T-cell development is how TCR occupancy can be perceived in such markedly different ways by the TCR. Here, we review the cytoplasmic and nuclear events that result from TCR signaling during thymocyte selection. Studies aimed at distinguishing molecular components involved in positive selection (resulting in signals for further differentiation) and negative selection (resulting in apoptosis) will help solve this fascinating feature of T-lymphocyte biology. We also discuss how non-TCR-derived signaling might serve to fine tune the TCR-driven selection events in thymocytes. Central to this aspect of the conceptual framework needed to explain thymocyte selection is the observation that thymic antigen-presenting cells appear to be specialized in the induction of either positive or negative selection. Finally, we suggest a hypothesis that integrates the facts currently available on developing thymocytes, and which may serve to refine our exploration of unresolved issues in thymocyte selection. This hypothesis expands our focus to include signals from receptors other than TCRs as modulating and amplifying factors in thymocyte signaling. PMID- 9850864 TI - The role of B7 co-stimulation in activation and differentiation of CD4+ and CD8+ T cells. AB - The functional significance of B7 co-stimulation in T-cell activation was described first in the context of preventing the induction of anergy. The functions of this pathway are far more complex than initially appreciated in view of the existence of two B7 molecules which have specificities for both CD28 and CTLA-4, which serve to amplify and terminate T-cell responses respectively. Mice lacking B7 co-stimulators and CD28 and CTLA-4 co-stimulatory receptors are helping to clarify the functions of this key immunoregulatory pathway. In this review we will focus on the role of B7 co-stimulation in the activation and differentiation of CD4+ helper cells and CD8+ cytotoxic cells. The contribution of B7 co-stimulation to CD4+ responses depends upon the activation history of the T-cell and the strength of the T-cell antigen receptor signal. B7 co-stimulation contributes to interleukin (IL)-2 production by both naive and previously activated CD4+ T cells. B7 co-stimulation is most critical for the differentiation of naive CD4+ T cells to IL-4 producers, but predominately influences IL-2 production by previously activated CD4+ cells. B7 co-stimulation is important in development of cytotoxic T cells through both effects on T-helper cells and by direct co-stimulation of CD8+ cells. PMID- 9850865 TI - Probing the activation requirements for naive CD8+ T cells with Drosophila cell transfectants as antigen presenting cells. AB - Activation of T cells involves multiple receptor-ligand interactions between T cells and antigen presenting cells (APC). At least two signals are required for T cell activation: Signal 1 results from recognition of MHC/peptide complexes on the APC by cell surface T-cell receptors (TCR), whereas Signal 2 is induced by the interactions of co-stimulatory molecules on APC with their complementary receptors on T cells. This review focuses on our attempts to understand these various signals in a model system involving the 2C TCR. The structural basis of Signal 1 was investigated by determining the crystal structure of 2C TCR alone and in complex with MHC/peptide. Analysis of these structures has provided some basic rules for how TCR and MHC/peptide interact; however, the critical question of how this interaction transduces Signal 1 to T cells remains unclear. The effects of Signal 1 and Signal 2 on T-cell activation were examined with naive T cells from the 2C TCR transgenic mice, defined peptides as antigen and transfected Drosophila cells as APC. The results suggest that, except under extreme conditions, Signal 1 alone is unable to activate naive CD8 T cells despite the induction of marked TCR downregulation. Either B7 or intercellular adhesion molecule (ICAM)-1 can provide the second signal for CD8 T-cell activation. However, especially at low MHC/peptide densities, optimal activation and differentiation of CD8 T cells required interaction with both B7 and ICAM-1 on the same APC. Thus, the data suggest that at least two qualitatively different co-stimulation signals are required for full activation of CD8 T cells under physiological conditions. PMID- 9850866 TI - Induction of peripheral CD8+ T-cell tolerance by cross-presentation of self antigens. AB - There is now convincing evidence that CD8+ T cells can be activated by professional antigen-presenting cells which present antigens derived from non lymphoid tissues in association with MHC class I molecules in the draining lymph nodes. This mechanism, referred to as cross-presentation, enables the immune system to respond to those microorganisms that infect only non-lymphoid tissues. Consistent with this view, cross-presentation was found to focus on antigens expressed in high concentrations and those released from dying cells, which can be expected to result from viral infections. Recent evidence, however, demonstrates that high dose self antigens can be cross-presented constitutively, resulting in the activation of autoreactive CD8+ T cells. This does not lead to auto immunity under physiologic conditions, but to CD95-mediated deletion of the T cells. Cross-presentation can thus engage a well-defined pathway of antigen induced T-cell death and purge the immune system of autoreactive CD8+ T cells. Low dose self antigens are not cross-presented and are consequently ignored. The immune system therefore uses two strategies to avoid CD8+ T-cell-mediated autoimmunity in the periphery: deletion of autoreactive CD8+ T cells responding to high dose self antigens and ignorance of self antigens expressed at low concentrations. PMID- 9850867 TI - T-cell survival. AB - Like other cells, T cells are dependent on signals from their environment for their survival. Resting T cells are supported in vitro by cytokines such as interleukin (IL)-4, IL-6 and IL-7. The latter two cytokines are made constitutively in animals and hence might affect the lifetimes of their resting T cells. Resting T cells are also kept alive by interaction with an as yet unidentified molecule on the surface of other cells. Activated T cells are also supported in vitro by members of two families of these proteins, the IL-2 family and the interferon-alpha beta family. Members of the latter family may have effects on activated cells in vivo. Thus although both resting and activated T cells require signals to keep themselves alive, the signals are different for the two types of cells. This perhaps allows the immune response to control the numbers of activated cells during infections without compromising its pool of precursor, resting T cells. PMID- 9850868 TI - Molecular regulation of interleukin-2 expression by CD28 co-stimulation and anergy. AB - The consequences of T-cell receptor engagement (signal 1) are profoundly affected by the presence or absence of co-stimulation (signal 2). T-cell receptor (TCR) stimulation in the absence of CD28-mediated co-stimulation not only results in little interleukin (IL)-2 production, but induces a long lasting hyporesponsive state known as T-cell clonal anergy. The addition of CD28 ligation to signal 1, on the other hand, results in the production of copious amounts of IL-2. Our laboratory has utilized CD4+ Th 1 clones in an effort to understand the molecular events resulting in enhanced IL-2 production by co-stimulation and the inhibition of IL-2 production in anergy. Our current studies have focused on defining the post-transcriptional effects of CD28-enhanced IL-2 production. The data suggest that a major component of CD28's ability to regulate IL-2 production occurs at the level of message stability and involves the 3'-untranslated region of the message. In terms of anergy, our recent studies support the notion that it is not the result of TCR engagement in the absence of co-stimulation, but rather signal 1 in the absence of IL-2 receptor signaling and proliferation. Furthermore, T cell anergy appears to be an active negative state in which IL-2 production is inhibited both at the level of signal transduction and by cis-dominant repression at the level of the IL-2 promoter. PMID- 9850869 TI - The role of clonal anergy in the avoidance of autoimmunity: inactivation of autocrine growth without loss of effector function. AB - Exposure of mature CD4+ T cells in the peripheral immune system to peptide antigen/MHC complexes in the absence of a threat of infection induces tolerance to the antigen as a result of both a decreased clonal frequency (peripheral deletion) and the induction of proliferative unresponsiveness (clonal anergy) in the survivors. Interestingly, Th 1-like effector functions are not automatically blocked after the development of clonal anergy. Thus, anergic T cells have the capacity to mediate Th 1-like helper activities if allowed to accumulate to high frequency. In this article, we examine those factors important to the development of tolerance versus immunity against protein antigen, and speculate on the relationship that exists between effective peripheral tolerance induction and the avoidance of autoimmune disease. PMID- 9850870 TI - Autopsy in geriatric psychiatry. PMID- 9850871 TI - Low plasma vitamin C in Alzheimer patients despite an adequate diet. AB - OBJECTIVE: To compare the vitamin C and E plasma levels in patients with Alzheimer's disease (AD) and to assess the vitamin C intake and nutritional status. DESIGN: Case-control study. Four groups of sex- and age-matched subjects were compared: severe AD and moderate AD, in patients with moderate AD and controls. SETTING: Community and hospitalized patients in the region of Toulouse, France. PARTICIPANTS: Patients with dementia who fulfilled criteria for Alzheimer's disease: severe Alzheimer group (N = 20), Mini-Mental State Examination (MMSE) score range 0-9; moderate Alzheimer group (N = 24), MMSE 10 23; hospitalized Alzheimer group (N = 9), MMSE 10-23. Control group (N = 19), MMSE 24-30. MEASURES: Plasma vitamin E and C were quantified by HPLC fluorescence. Consumption of raw and cooked fruit and vegetables was evaluated in order to determine the mean vitamin C intakes. Mini Nutritional Assessment (MNA) and plasma albumin were used to measure nutritional status. RESULTS: Institutionalized and community subjects were analysed separately. MNA scores were normal in home-living Alzheimer subjects with moderate dementia and significantly lower in those with severe disease, despite normal plasma albumin levels. In the home-living Alzheimer subjects, vitamin C plasma levels decreased in proportion to the severity of the cognitive impairment despite similar vitamin C intakes, whereas vitamin E remained stable. The hospitalized Alzheimer subjects had lower MNA scores and albumin levels but normal vitamin C intakes, but their plasma vitamin C was lower than that of community-living subjects. Institutionalized Alzheimer subjects had significantly lower MNA scores but normal vitamin C and albumin levels and vitamin C intakes compared with community dwelling subjects of similar degree of cognitive impairment. CONCLUSION: Plasma vitamin C is lower in AD in proportion to the degree of cognitive impairment and is not explained by lower vitamin C intake. These results support the hypothesis that oxygen-free radicals may cause damage. PMID- 9850872 TI - Natural history of Alzheimer's disease and other dementias: review of the literature in the light of the findings from the Kungsholmen Project. AB - The elderly population is increasing more than any other sector of the population. Dementia, a prevalent condition in the elderly, increases disability, morbidity and mortality among older people. For these reasons the possibility of predicting progression and prognosis has enormous importance. Despite the fact that dementia has gained widespread recognition in the past few decades, the knowledge of its natural history, in terms of progression and prognosis are not yet completely understood. However, thanks to longitudinal research, which has only recently begun to proliferate, not only is better comprehension of the continuity of the cognitive decline possible, but also the identification of some prognostic factors. PMID- 9850873 TI - Apolipoprotein E epsilon 4 allele in association with global cognitive performance and CSF markers in Alzheimer's disease. AB - To better define the influence of apolipoprotein E (ApoE) epsilon 4 genotype on the cognitive and biochemical features of Alzheimer's disease (AD), cross sectional analysis of global cognitive measures and cerebrospinal fluid studies gathered on AD subjects at a tertiary care facility between 1986 and 1997 was carried out. The 112 AD patients examined included 62 women and 50 men with a mean (SD) age of 64.2 (9.2) years. Patient demographics; illness onset age and duration, education level and global cognitive measures were recorded systematically. Genetic analysis for ApoE allele type and biochemical characterization of CSF, including total tau concentration, was performed. Descriptive statistics of demographics, cognitive and CSF measures were performed by chi-square, ANOVA and Tukey's tests. Overrepresentation of the epsilon 4 allele was found, with 45.5% of AD patients heterozygous and 20.5% homozygous for ApoE epsilon 4. Overall, ApoE epsilon 4 status had no effect on mean onset age of AD (F = 1.56; p = 0.214), but an earlier mean onset age of AD (F = 4.10; p = 0.02) was seen in the late-onset subjects. No differences were found with regard to ApoE epsilon 4 status and measures of disease, duration of illness or global cognitive performance. Although CSF tau was elevated in our sample (575.4 +/- 290.3 pg/ml), ApoE epsilon 4 status did not influence total CSF tau or neurotransmitter metabolite levels. ApoE epsilon 4 genotype had no impact on a variety of illness severity, cognitive and CSF examinations in the largest cross sectional analysis of AD subjects yet reported. PMID- 9850874 TI - Late paraphrenia: a variant of schizophrenia manifest in late life or an organic clinical syndrome? A review of recent evidence. AB - A large amount of research has been devoted during the past 15 years to the clinical and neurobiological aspects of the disorder named as 'late paraphrenia' (LP) in 1955. The symptomatology and diagnosis of the disorder, its prognosis, the cognitive functioning of those affected, the structural changes in the brain as revealed by modern techniques of brain imaging and its postmortem neuropathology have all been submitted to investigation. The results have been widely regarded as consistent with the concept of LP as an organic disease of the brain, but increased knowledge of the neurobiology of schizophrenia and of the age-related changes that occur in the brains of elderly people casts doubt on the validity of this interpretation. The findings are consistent with the view that LP is the form in which schizophrenia is manifest in old age. The proposal that LP has a closer kinship with affective disorder than with schizophrenia is part of a general theory of the sex differences in schizophrenia. In LP it becomes entangled with the organicity hypothesis, suggesting that neither of these explanations is adequate, and most of the evidence points to a unitary concept which views LP as a variant of a single disorder, namely schizophrenia, which, however, requires a broad definition. This concept has implications for fresh paths of enquiry. PMID- 9850875 TI - Incidence and relationship between behavioural and psychological symptoms in Alzheimer's disease. AB - OBJECTIVES: To evaluate the frequency and type of psychological and behavioural symptoms in Alzheimer's disease (AD) patients in Poland, in various stages of the disease. METHOD: One hundred and sixty-nine patients with a diagnosis of probable AD in Global Deterioration (GDS) stages 3, 4, 5, 6 and 7 of dementia were examined in a search for behavioural and psychological symptoms. RESULTS: Behavioural and psychotic symptoms were most often found in GDS stages 5 and 6 of AD, except for depressive disorder, which was observed most frequently in GDS stage 4 and whose frequently decreased towards the terminal stages of dementia. From an analysis of the relationship between behavioural symptoms in the Polish AD patients, the following syndromes may be discriminated: psychotic syndrome (delusions and hallucinations), delusions with aggressive behaviour and hallucinations and anxiety. With more severe dementia, the syndromes, which could be the result of delirium, became more common. CONCLUSIONS: Diagnosis of delirium should be considered in moderately severe and severe dementia whenever a sudden change in patients' behaviour occurs. PMID- 9850876 TI - The behavioral pathology in Alzheimer's Disease Scale (BEHAVE-AD): factor structure among community-dwelling Alzheimer's disease patients. AB - OBJECTIVE: The aims of this study were to (a) determine the factor structure of the Behavioral Pathology in Alzheimer's Disease Scale (BEHAVE-AD), and (b) examine the associations of the observed factors to the level of cognitive impairment. DESIGN: Cross-sectional study of geriatric patients evaluated at an outpatient memory disorders clinic. SAMPLE: One hundred and fifty-one consecutive patients diagnosed with Alzheimer's disease (AD) according to NINCDS-ADRDA diagnostic criteria. RESULTS: Principal factors analysis with Varimax rotation resulted in a five-factor solution that accounted for 40.0% of the common variance. The factors included agitation/anxiety (agitation, anxiety of upcoming events; other anxiety), psychosis (delusions of theft, suspiciousness/paranoia; visual hallucinations), aggression (verbal aggressiveness; physical threats/violence; fear of being left alone; other delusions), depression (tearfulness; depressed mood) and activity disturbance (wandering; delusion one's house is not one's home). Several factors were associated with level of cognitive impairment as assessed by the Mini-Mental State Examination (MMSE). CONCLUSION: The results of this study suggest that the BEHAVE-AD measures a wide range of behavioral pathology that can be empirically represented by five independent symptom clusters among outpatient AD patients. PMID- 9850877 TI - Social determinants of psychiatric morbidity and well-being in immigrant elders and whites in east London. AB - OBJECTIVES: The social conditions under which migrants to the UK live may be more significant than the experience of migration itself in leading to increased risk of mental illness. We aimed to compare the prevalence of mental, physical and social health problems in elderly Somalis, Bengalis and whites living in a deprived inner London area and examine associations between environmental circumstances, social support, physical health status, mood and life satisfaction in these groups. In addition, we wanted to test the hypothesis that differences in mental health between immigrants and whites are explained by social disadvantages rather than ethnicity. DESIGN: Cross-sectional survey with participants drawn from age-sex registers of general practices, augmented by other sources. SETTING: East London--'first-generation' Somali and Bengali immigrants and white British. SUBJECTS: A total of 274 people aged 60+ years: 72 Somalis, 75 Bengalis and 127 whites. MAIN OUTCOME MEASURES: Symptoms of Anxiety and Depression Scale (SAD), Life Satisfaction Index (LSI). High SAD scores indicate more anxiety and depression symptoms; high LSI scores indicate greater life satisfaction. MAIN RESULTS: Highest SAD scores were found among Bengalis; lowest LSI scores were found among Bengalis and Somalis. The prevalences of depression (SAD score 6+) were 25% in Somalis, 77% in Bengalis and 25% in east London whites. Physical health status and SAD scores were associated in Somalis (r = +0.31, p < or = 0.01). Bengalis (r = +0.47, p < or = 0.001) and east London whites (r = +0.27, p < or = 0.01). Physical health problems also related to lower LSI scores in Somalis (r = -0.24, p < or = 0.05) and east London whites (r = 0.24, p < or = 0.01). Social factors (i.e. poor housing conditions, low family support and reported need of community services) were strongly associated with SAD scores among Somalis (r = +0.5, p < or = 0.001) and, to a lesser extent, among Bengalis (r = +0.33, p < or = 0.01). Ethnicity (i.e. being an immigrant as opposed to a non-immigrant) became a statistically non-significant risk factor for high SAD scores after adjusting for the effects of age, weekly income, physical health and social problems (OR = 0.71, 95% CI = 0.5-1.1, p = 0.09). A residual, but much attenuated effect for ethnicity on LSI scores persisted in the estimated model after controlling for the same set of independent risk factors (OR = 0.7, 95% CI = 0.4-1, p = 0.05). CONCLUSION: The marked variation in mental health between ethnic groups in east London might be a reflection of socioeconomic and health differentials acting concomitantly and adversely. Inequalities in housing, social support, income and physical health status accounted for variation in mood observed between immigrants and whites, and may partly explain differences in life satisfaction. These results seem to support a 'multiple jeopardy' theory of ageing in ethnic minorities in east London. Greater efforts are needed to recognize anxiety and depression in immigrant elders. Better social support and housing among 'minority ethnic' elders who live alone might be expected to alleviate social stress and improve mental health and psychological well-being. PMID- 9850878 TI - Recent life events as major predictors of mental health in the oldest old. PMID- 9850879 TI - Mania associated with donepezil. PMID- 9850880 TI - Hynonatremia in elderly psychiatric inpatients. PMID- 9850881 TI - Role of iron in Alzheimer-type dementia in Down syndrome. PMID- 9850882 TI - Re: Geriatric consultation and liaison service. PMID- 9850883 TI - Current awareness in geriatric psychiatry. PMID- 9850884 TI - Reporting astigmatism data. PMID- 9850885 TI - Simplicity and complexity in topical anesthesia for cataract surgery. PMID- 9850886 TI - Assessment of surgically induced astigmatism: toward an international standard. PMID- 9850887 TI - Format for reporting surgically induced astigmatism on aggregate data. PMID- 9850888 TI - Assessment of surgically induced astigmatism: toward an international standard II. PMID- 9850889 TI - Consultation section. Cataract surgical problem. PMID- 9850890 TI - Simplified anesthesia technique for scleral tunnel phacoemulsification. AB - A simplified anesthesia technique for sutureless scleral tunnel, small incision, one-port phacoemulsification is described. Step 1: Mild intravenous sedation is achieved with 1 mg of midazolam and 10 mg of propofol. Step 2: A tetracaine- or preservative-free-lidocaine-soaked Weck cell sponge is applied to the superior bulbar conjunctiva and the superior episcleral tissue for localized anesthesia. Step 3: Preservative-free lidocaine 1%, 0.75 cc, is injected into the anterior chamber for intraocular anesthesia. This technique can be used in the presence of all types of cataracts, requires about 10 seconds to complete, and can be used with all phacoemulsification techniques. Results of the technique in a study population are reported. PMID- 9850891 TI - Relative strength of photodynamic biologic tissue glue in penetrating keratoplasty in cadaver eyes. AB - PURPOSE: To compare the relative strength of photodynamic biologic tissue glue (PBTG) with that of 10-0 nylon sutures in closing penetrating keratoplasty (PKP) wounds in cadaver eyes. SETTING: Corneal Research Laboratory, University of Chicago, Chicago, Illinois, USA. METHODS: A 7 mm central corneal trephination was performed on 8 recently enucleated human eyes. Each corneal button was removed and then resutured in its original position, using 4 interrupted 10-0 nylon cardinal sutures and PBTG (n = 7) or 16 interrupted 10-0 nylon sutures (n = 1). Wound leak and incisional bursting pressures were determined. RESULTS: The average pressure at which wound leak and iris prolapse occurred was 124 mm Hg (range 70 to 180) and 185 mm Hg (range 90 to 300), respectively. The pressure required to create wound leak increased as the time between glue application and pressurization of the globe lengthened (mean 22 minutes, range 5 to 60 minutes), indicating that the bonding and stability of the adhesive increased with time. In contrast, the control eye developed wound leak at 70 mm Hg and iris prolapse at 300 mm Hg. CONCLUSIONS: Photodynamic biologic tissue glue was comparable to sutures in providing adequate corneal wound strength in PKP in a cadaver eye model. Wound closure with PBTG may reduce the number of sutures required in corneal transplantation and decrease the incidence of suture-related complications and allograft rejection. PMID- 9850892 TI - Changes in corneal epithelial barrier function after excimer laser photorefractive keratectomy. AB - PURPOSE: To use fluorophotometry to measure corneal epithelial barrier function after excimer laser photorefractive keratectomy (PRK). SETTING: Seoul National University Hospital, Seoul, Korea. METHODS: Twenty-five eyes of 21 patients (13 women, 8 men) had PRK to correct myopia. Corneal epithelial healing time was measured and corneal epithelial permeability to sodium fluorescein evaluated by fluorophotometry 1, 2, and 3 weeks after surgery. RESULTS: Epithelial permeability showed a statistically significant increase 1 week after surgery and returned to its preoperative level 1 week later. Comparative studies according to epithelial healing day and corrected diopter showed results that were not statistically significant (P > .05). CONCLUSION: These results suggest that PRK delays complete reconstruction of corneal epithelial barrier function. In humans, the corneal epithelium regained its normal barrier function 2 weeks after PRK. Thus, at least during these weeks, care should be taken to minimize further epithelial trauma. PMID- 9850893 TI - Corneal hydration and central islands after excimer laser photorefractive keratectomy. AB - PURPOSE: To determine whether uneven corneal surface hydration during excimer laser photorefractive keratectomy (PRK) is related to postoperative occurrence of central islands. SETTING: LSU Eye Center, New Orleans, Louisiana, USA. METHODS: A retrospective study reviewed intraoperative videotapes and postoperative videokeratography of 49 eyes of 49 patients who had excimer laser PRK for myopia. The uniformity of corneal hydration within the photoablation zone, particularly the frosty appearance of the ablated zone, was characterized. The presence or absence of a topographic central island (steepening of at least 3.0 diopters and 1.5 mm in diameter) was determined from the 1 month postoperative videokeratographs. RESULTS: Twelve eyes (24.5%) developed central islands postoperatively. A statistically significant association was observed between the uneven surface hydration (central accumulation of fluid) within the ablation zone intraoperatively and the formation of central islands postoperatively (P < .001, Kruskal-Wallis test; Kendall tau rank correlation = 0.534; P < .001). CONCLUSION: Nonuniform fluid distribution during photoablation was a risk factor for central island formation after PRK. Intraoperatively, the presence of excess fluid in the central cornea appeared as a shiny area. This mirror-like surface layer may reduce the rate of central ablation by reflecting and absorbing a significant amount of the incident excimer laser light. PMID- 9850894 TI - Three year results of photoastigmatic refractive keratectomy for mild and atypical keratoconus. AB - PURPOSE: To evaluate the long-term effect of photoastigmatic refractive keratectomy (PARK) on mild keratoconus. SETTING: Laser Unit, Ein Tal Eye Center, Tel Aviv, Israel. METHODS: Eight eyes of 6 patients with stable compound myopic astigmatism and topography features of keratoconus were treated with a VISX Twenty-Twenty excimer laser. The laser beam slit width ranged between 4.8 and 5.0 mm according to the degree of cylinder. Preoperative slitlamp examination did not reveal significant thinning, ectasia, or scarring. Astigmatism ranged between 2.00 and -7.50 diopters and follow-up, between 36 and 48 months. RESULTS: In 7 of 8 eyes, uncorrected visual acuity (UCVA) before PARK ranged between 6/30 and finger counting. After PARK, it ranged between 6/9(-) and 6/15, and patients could manage in most of their daily activities without using spectacles. In 2 eyes, the final UCVA was 6/15(-) and 6/15. In 1 eye, treatment failed to improve UCVA and corneal topography revealed progression of the keratoconus. CONCLUSIONS: Photoastigmatic keratectomy was partially effective in eyes with mild stable keratoconus. However, the long-term results may be altered by progression of corneal thinning and ecstatic disease. In addition to the mild nature of the keratoconus, this is a small series and further studies are needed to confirm these results. PMID- 9850895 TI - Interface inflammation after laser in situ keratomileusis. Sands of the Sahara syndrome. AB - PURPOSE: To determine the source of the interface debris that causes the interface inflammation known as "sands of the Sahara" after laser in situ keratomileusis (LASIK). SETTING: Department of Ophthalmology, LSU Eye Center, Louisiana State University Medical Center School of Medicine, New Orleans, USA. METHODS: A microkeratome (Automated Corneal Shaper) was used to make a LASIK flap in 8 eyes of 4 rabbits. In 4 eyes, the blade was used directly from the sterile pack; in the contralateral 4 eyes, the blade was cleaned prior to use. In vivo confocal microscopy of the corneas was performed 1 day after surgery. An unused, cleaned blade and an unused, uncleaned blade, as well as blades used in the rabbit eyes, were examined by scanning electron microscopy. RESULTS: Confocal microscopy revealed numerous fragments of debris surrounded by inflammatory cells in the LASIK flap interfaces created by blades taken directly from the sterile package. Interfaces created by the cleaned blades showed only rare, scattered bits of debris. Scanning electron microscopy of the unused blades showed debris on the uncleaned blade removed directly from the sterile package. CONCLUSION: Post-LASIK interface inflammation may be caused by debris on the microkeratome blade, although other sources are possible. The interface debris and inflammation can be reduced or eliminated by cleaning the microkeratome blade before use. PMID- 9850896 TI - Preoperative measurement of scotopic pupil dilation using an office pupillometer. AB - PURPOSE: To describe a light-amplification pupillometer designed for office evaluation of refractive surgical candidates and compare its usefulness with that of a millimeter ruler and a standard pocket-card pupillary gauge. SETTING: Center for Ophthalmic Surgery, Encino, California, USA. METHOD: The pupil sizes of 100 patients with a mean age of 28 years (range 18 to 50 years) were measured at 2 levels of illumination: 15 and 3 lumens. Pupil size was measured with a Rosenbaum pocket-card pupil gauge, a millimeter ruler, and the pupillometer at approximately 15 lumens. This level of illumination was chosen by 3 technicians as the lowest at which the pupil of a brown eye could be measured with confidence using the Rosenbaum card. Pupil size was then measured using the light amplification pupillometer at approximately 3 lumens. This illumination level was chosen to simulate that of night-driving conditions. RESULTS: At 15 lumens, mean pupil diameter was 5.1 mm (range 2.5 to 8.0 mm). There was no difference in the measurements taken with the 3 instruments at this illumination level. At 3 lumens, the pupil diameter could not be measured with confidence using the Rosenbaum card or the millimeter ruler. Using the pupillometer, mean pupil size was 6.2 mm (range 3.0 to 9.0 mm). The mean difference in pupil diameter measured at 15 and 3 lumens was 1.1 mm (range 0.0 to 3.0 mm). CONCLUSION: This evaluation suggests that it would be difficult for clinicians to reliably predict the level of pupil dilation at 3 lumens by measuring pupil size at 15 lumens. The light amplification pupillometer provides a tool for measuring pupil size at an illumination level that simulates night-driving conditions. The use of this device may help surgeons identify refractive surgery candidates who are more likely to be dissatisfied with their postoperative vision at low illumination levels. PMID- 9850897 TI - Aqueous humor lidocaine concentrations in topical and intracameral anesthesia. AB - PURPOSE: To assess aqueous humor lidocaine concentrations in 2 common regimens of topical anesthesia and after intracameral injection of the anesthetic agent. SETTING: University hospital eye clinic. METHODS: Twenty patients having routine cataract surgery were randomized into 3 groups: 1 given 3 drops of lidocaine 4% before surgery; 1 given 6 drops; 1 given 3 drops plus an intracameral injection of 0.1 mL lidocaine 1%. Lidocaine concentration was measured in aqueous humor samples taken before surgery. RESULTS: With 3 drops, aqueous lidocaine concentration was 1.4 micrograms/mL +/- 0.5 (SD) and with 6 drops, 4.3 +/- 1.5 micrograms/mL (P = .0015). With an intracameral injection, it was 341.8 +/- 152.6 micrograms/mL. CONCLUSION: Measurable amounts of lidocaine entered the anterior chamber in topical anesthesia, and more entered when more drops were given. It is likely that concentrations in this range could anesthetize the iris, but they are far lower than concentrations after an intracameral injection. PMID- 9850898 TI - Topical tetracaine versus topical tetracaine plus intracameral lidocaine for cataract surgery. AB - PURPOSE: To compare topical tetracaine 0.5% alone and with intracameral lidocaine 1% as a local anesthetic agent in phacoemulsification with intraocular lens (IOL) implantation. SETTING: The Toronto Hospital-Western Division, Toronto, Canada. METHODS: Fifty-nine consecutive patients (60 eyes) having phacoemulsification with implantation of a foldable acrylic IOL (AcrySof) were randomized into 1 of 2 groups: The intracameral balanced salt solution (BSS) group received topical tetracaine 0.5% plus intracameral BSS; the intracameral lidocaine group received topical tetracaine 0.5% with preservative-free intracameral lidocaine 1%. The patients' subjective experience of pain was measured at 4 points during surgery using a 4-point pain scale. Patient and surgeon satisfaction with the anesthesia used was measured using a 5-point satisfaction scale. Central endothelial cell counts were obtained preoperatively and 1 month postoperatively. Best corrected visual acuity (BCVA) was measured preoperatively and 1 hour, 1 day, 1 week, and 1 month postoperatively. RESULTS: The mean pain score after phacoemulsification was significantly higher in the intracameral BSS group than in the intracameral lidocaine group (0.63 +/- 0.7 [SD] and 0.23 +/- 0.4, respectively, P < .019). The mean pain score at the end of surgery was also significantly higher in the intracameral BSS group than in the intracameral lidocaine group (0.60 +/- 0.6 and 0.21 +/- 0.4, respectively; P < .014). The surgeon satisfaction score was significantly lower for the intracameral BSS group than for the intracameral lidocaine group (3.90 +/- 1.2 and 4.73 +/- 0.8, respectively; P < .0007). There was no difference in patient satisfaction between the intracameral BSS and intracameral lidocaine groups (4.60 +/- 0.6 and 4.70 +/- 0.8). Endothelial cell loss 1 month postoperatively was similar between the 2 groups (6.1% +/- 8% and 6.7% +/- 6%). Ninety-seven percent of patients (29/30) in each group noted BCVA improvement from preoperatively. The rate of potential visual acuity recovery was similar in both groups. CONCLUSION: Topical tetracaine 0.5% with intracameral lidocaine was safe and effective in patients having phacoemulsification with IOL implantation. The advantage of using intracameral lidocaine 1% over a placebo was a significant decrease in the patients' subjective experience of pain and in the surgeon's satisfaction with the anesthesia used. None of the other parameters measured in this study differed significantly between the 2 groups. PMID- 9850899 TI - Pain in scleral pocket incision cataract surgery using topical and peribulbar anesthesia. AB - PURPOSE: To evaluate the pain produced during different phases of phacoemulsification cataract surgery using a scleral pocket incision under topical versus peribulbar anesthesia. SETTING: Department of Ophthalmology, Oulu University Hospital, Oulu, Finland. METHODS: This prospective study comprised 100 cataract patients who were randomly selected to have phacoemulsification with a scleral pocket incision using either topical or peribulbar anesthesia. Topical anesthesia comprised oxybuprocaine 0.4% drops. Peribulbar anesthesia was given with an inferolateral transconjuctival injection of an even mixture of lidocaine 2% and bupivacaine 0.5% with hyaluronidase. Inadvertent eye movement during surgery was recorded. Pain occurring during intravenous line cannulation, introduction of the anesthetic agent, and phacoemulsification was measured using a visual analog scale (from 0 to 10) and a descriptive verbal 5-step scale. Patients were asked about pain immediately after each phase. RESULTS: The pain during cannulation was similar in both groups (P = .498). The peribulbar injection was statistically significantly more painful than induction of topical anesthesia (2.11 and 0.10, respectively; P < .001). Surgery was statistically significantly more painful in the topical group than in the peribulbar group (2.76 and 0.85, respectively; P < .001). The mean pain score during all 3 phases was similar (1.43 topical group and 1.51 peribulbar group; P = .500). On the verbal scale, surgery was more painful under topical than under peribulbar anesthesia (P < .001). There were no statistical differences in pain during the peribulbar injection and during cannulation (P = .461 and P = .462, respectively). Inadvertent eye movement occurred more often in the topical anesthesia group. CONCLUSION: Considering the entire procedure, total pain using topical anesthesia was acceptable and equal to that using peribulbar anesthesia for phacoemulsification with a scleral pocket incision. Pain during phacoemulsification was greater under topical anesthesia but not significantly different from the pain during the peribulbar injection. PMID- 9850900 TI - Inhibition of posterior capsule opacification with an immunotoxin specific for lens epithelial cells: 24 month clinical results. AB - PURPOSE: To assess the safety and effectiveness of an immunotoxin, MDX-RA, designed to inhibit posterior capsule opacification (PCO). SETTING: Eleven private practices in the United States. METHODS: This study comprised 63 eyes of 63 patients having extracapsular cataract extraction by phacoemulsification; these patients were enrolled in a Phase I/II clinical investigation of the immunotoxin MDX-RA. At the close of surgery, 21 patients were treated with placebo, 23 patients with 50 units of the immunotoxin, and 19 patients with 175 units of the immunotoxin as an aqueous solution. The patients were monitored for 24 months after primary cataract surgery using external eye and slitlamp examinations, visual acuity assessment, ophthalmoscopy, pachymetry, tonometry, endothelial cell counts, and lens capsule photography. Posterior capsule opacification, recorded on lens capsule photographs, was graded independently by a committee of 3 cataract surgeons. The incidence of neodymium:YAG (Nd:YAG) capsulotomy was projected from the opacification results. RESULTS: The immunotoxin, at the 50 unit dose, was well tolerated and effective in inhibiting PCO. At the 175 unit dose, there was a trend toward increased postoperative inflammation that was transient with no residua. From 6 to 24 months postoperatively, the 50 unit dose significantly inhibited PCO compared with the placebo (P < .05). This significant reduction in PCO translated into a significantly lower projected need for Nd:YAG capsulotomy in the 50 unit than the placebo group (P < .004). About 60% in the placebo group and 4% in the 50 unit group were projected to need an Nd:YAG capsulotomy by 3 years postoperatively. CONCLUSION: The immunotoxin was well tolerated and was effective in reducing PCO for up to 24 months after cataract surgery. Although these preliminary results are encouraging, a larger study is underway to determine whether the reduction in PCO by the immunotoxin decreases the need for Nd:YAG capsulotomy. PMID- 9850901 TI - Mathematical model to predict the need for neodymium: YAG capsulotomy based on posterior capsule opacification rate. AB - PURPOSE: To evaluate a model to project the estimated time required before patients having primary phacoemulsification require neodymium:YAG (Nd:YAG) laser capsulotomy. SETTING: Eleven private practices in the United States. METHODS: Projections of time to capsulotomy were based on assessment of the early development of posterior capsule opacification (PCO) over time. The PCO data were collected during a clinical study to evaluate MDX-RA, an investigational immunotoxin designed to limit epithelial cell growth, preventing postsurgical PCO. From the PCO data, the estimated time to Nd:YAG capsulotomy in a placebo treated group was compared with the actual time to capsulotomy in a cohort of patients from general practice who had had phacoemulsification. RESULTS: By 6 months, the mean Opacification Index in the MDX-RA group was significantly lower than that in the placebo group (P < .05) and it remained significantly lower at 12 (P < .001), 18 (P < .001), and 24 (P < .016) months. The rate of PCO in the MDX-RA group was approximately 6 times lower than that in the placebo group (P < .0004). Fifty-seven percent in the placebo group and 4% in the MDX-RA group were projected to require an Nd:YAG capsulotomy within 3 years of primary cataract surgery. Projected values for the placebo group were similar to actual values observed in the population-based cohort. CONCLUSIONS: This technique could be used to predict the need for Nd:YAG capsulotomy using early measurements of PCO. PMID- 9850902 TI - Comparison of methods to assess visual impairment from glare and light scattering with posterior capsule opacification. AB - PURPOSE: To compare 2 glare tests to determine their relative usefulness in the assessment of posterior capsule opacification (PCO) and to evaluate the potential benefits of combined visual, acuity, contrast sensitivity, and glare testing. SETTING: Teaching hospital ophthalmology department. METHODS: Sixteen patients had glare, visual acuity, and contrast sensitivity testing before and after neodymium:YAG (Nd:YAG) capsulotomy. Results with the Brightness Acuity Tester (BAT, Mentor), which measures disability glare, and the Straylightmeter (Foundation for Eye Research, The Netherlands), which quantifies forward scatter by direct compensation techniques, were compared. The correlation between glare, ETDRS visual acuity, and Pelli-Robson contrast sensitivity was determined. RESULTS: Pretreatment visual acuity was significantly correlated with contrast sensitivity (P < .01). However, visual acuity and contrast sensitivity were poorly correlated with both the BAT and Straylightmeter (P > .05), indicating that visual acuity is predictive of contrast sensitivity but a poor predictor of glare. Glare was significantly improved (Straylightmeter, P < .0001; BAT, P < .05) following capsulotomy. While the Straylightmeter consistently measured precapsulotomy forward scatter that improved with treatment, corresponding BAT disability glare was unmeasurable in 18.8% of patients with PCO, as their visual acuities improved rather than deteriorated with glare testing. CONCLUSIONS: Glare testing provided more information than contrast sensitivity when combined with visual acuity in the evaluation of PCO. Glare related to PCO is better assessed using the Straylightmeter because the BAT may yield aberrant disability glare results. PMID- 9850903 TI - Reproducibility of posterior capsule opacification measurement using Scheimpflug videophotography. AB - PURPOSE: To examine the intraobserver and interobserver reproducibility in measuring posterior capsule opacification (PCO) using a Scheimpflug videophotography system. SETTING: Hayashi Eye Hospital, Fukuoka, Japan. METHOD: Twenty eyes with PCO and 10 without PCO had 5 independent measurements 15 minutes apart performed by 3 observers. The eyes without PCO were selected within 1 week after surgery. Intraobserver and interobserver reproducibility were evaluated using the coefficient of variation and intraclass correlation coefficient. RESULTS: The median coefficients of variation of the 3 observers were 11.6%, 11.3%, and 9.8%, respectively, which indicates an acceptably low observer variability. The intraobserver correlation coefficients of the 3 observers were all more than 0.95, and the interobserver correlation coefficient was 0.976, which indicates excellent reproducibility. CONCLUSION: The results of intraobserver and interobserver reproducibility of PCO measurement using the Scheimpflug videophotography system indicate it would be effective for use in research and clinical management of PCO. PMID- 9850904 TI - Surgically induced astigmatism with superior and temporal incisions in cases of with-the-rule preoperative astigmatism. AB - PURPOSE: To evaluate surgically induced astigmatism (SIA), postoperative astigmatism, and uncorrected visual acuity (UCVA) after cataract surgery with superior corneal, superior scleral, and temporal corneal 4.0 mm sutureless incisions in cases of preoperative with-the-rule (WTR) astigmatism. SETTING: Hopital des Quinze-Vingts, Paris, France. METHODS: This prospective evaluation included patients having phacoemulsification with foldable lens implantation through a 4.0 mm incision. Patients with preoperative WTR astigmatism were randomly assigned to 1 of 3 incisions: superior corneal (Group 1), superior scleral (Group 2), or temporal corneal (Group 3). All patients had autokeratometry preoperatively and postoperatively (1 day, 1 week, 1 month, 1 year). Surgically induced astigmatism using the vector method, postoperative astigmatism, and UCVA (patients whose spherical equivalent was with +/- 1 diopter) were evaluated. RESULTS: Ninety patients were included in the study; there were 30 in each incision group. One year postoperatively, Group 1 had 1.52 diopters (D) of SIA and 1.36 D of postoperative astigmatism; 53.5% of patients had a UCVA of 20/32 or better, Group 2 had 0.69 D of SIA (P < .05) and 0.67 D of postoperative astigmatism (P < .05); 82.7% of patients had a UCVA of 20/32 or better (P < .05). Group 3 had 0.69 D (P > .05), 0.98 D (P < .05), and 79.3% (P > .05), respectively. CONCLUSIONS: In this study, the superior corneal incision produced significant SIA, leading to high postoperative astigmatism and poor UCVA. The scleral and temporal incisions produced minimal SIA and good UCVA. PMID- 9850905 TI - Long-term course of surgically induced astigmatism after a 5.0 mm sclerocorneal valve incision. AB - PURPOSE: To study surgically induced astigmatism (SIA) after a sutureless 5.0 mm sclerocorneal valve incision. SETTING: Department of Ophthalmology, University of Vienna, Vienna, Austria. METHODS: This study evaluated SIA in 34 cases of sutureless cataract surgery with a 5.0 mm superior sclerocorneal valve incision and implantation of a poly(methyl methacrylate) intraocular lens. Keratometry was measured with a Zeiss keratometer in all cases preoperatively, and 1 day, 1 week, 1 and 3 months, and 1 and 5 years postoperatively. RESULTS: Surgically induced astigmatism, calculated by Cravy's vector analysis, showed an initial mean with the-rule shift of 0.35 diopter (D), followed by an against-the-rule shift to a mean of -0.30 D after 1 month. One year postoperatively, mean SIA was -0.46 D. Between 1 and 5 years postoperatively, there was a statistically significant increase in mean SIA calculated by Cravy's vector analysis; SIA increased from 0.46 D after 1 year to -0.76 D after 5 years postoperatively. CONCLUSIONS: A small, although statistically significant, amount of postoperatively induced astigmatism occurred 5 years after a sutureless 5.0 mm sclerocorneal valve incision. PMID- 9850906 TI - Two year follow-up of astigmatism after phacoemulsification with adjusted and unadjusted sutured versus sutureless 5.2 mm superior scleral incisions. AB - PURPOSE: To evaluate changes in astigmatism 6 months to 2 years after 5.2 mm superior scleral incision phacoemulsification using 3 closures. SETTING: Department of Ophthalmology, Vejle Hospital, Vejle, Denmark. METHODS: This long term follow-up study included 75 consecutive phacoemulsification patients who were randomly allocated to 1 of 3 incision closures: 1 intraoperatively adjusted cross suture, 1 unadjusted cross suture, no suture. Postoperative astigmatism after 1 and 2 years was evaluated by keratometric cylinder, induced astigmatism (Naeser's polar values), induced cylinder (Jaffe's vector analysis), and vector decomposition (Olsen). The data were compared with 1 week values. RESULTS: In the previous study, median astigmatism after 6 months was similar in all 3 groups, but only the sutureless group showed early stability. Significant against-the rule (ATR) changes were seen in the sutured cases (P < .01). The 1 and 2 year follow-up showed continued stability in the sutureless cases. The adjusted-suture group exhibited stability after 6 months, while the unadjusted-suture group showed a further tendency to change ATR. At 2 years, median ranges of astigmatism were -0.49 to -0.90 diopter (D) (Naeser), 0.77 to 1.02 D (Jaffe), and 96% to 98% ATR (Olsen). The intergroup differences were not statistically significant. CONCLUSION: Two year follow-up confirmed that sutureless closure led to early astigmatism stability. Using a suture prolonged the postoperative period of astigmatism instability, and if used without intraoperative adjustment, a tendency toward a less predictable outcome persisted after 2 years. PMID- 9850907 TI - Position of poly(methyl methacrylate) and silicone intraocular lenses after phacoemulsification. AB - PURPOSE: To compare the positional change of poly(methyl methacrylate) (PMMA) and silicone intraocular lenses (IOLs) in vivo after phacoemulsification. SETTING: Taipei Municipal Yang-Ming Hospital, Taipei, Taiwan. METHODS: A prospective study of 70 cataractous eyes treated by phacoemulsification with IOL implantation was carried out. The eyes were randomized into 2 groups based on IOL type: 1-piece PMMA IOL; 3-piece silicone IOL. The amount of IOL tilt and decentration was measured and anterior chamber depth (ACD) determined by Scheimpflug photography using an anterior eye segment analysis system (EAS-1000, Nidek). All eyes were examined 1 week and 1 to 6 months after surgery. RESULTS: No statistically significantly differences were found in the amount of tilt and decentration between 2 IOL types throughout the study. The ACDs were relatively constant in both groups through the early postoperative periods. CONCLUSION: The stability of PMMA and silicone IOLs were the same after phacoemulsification. PMID- 9850909 TI - Cataract formation after posterior chamber phakic intraocular lens implantation. AB - Posterior chamber phakic intraocular lens (PCP IOL) implantation is an emerging refractive procedure. We report a case of cataract formation 6 months after uneventful implantation of a Staar PCP IOL to correct high myopia. Visual recovery was achieved after explantation of the phakic IOL and phacoemulsification with implantation of a foldable IOL through the same unenlarged self-sealing corneal incision. PMID- 9850908 TI - Combined topical-peribulbar anesthesia for cataract surgery. AB - PURPOSE: To evaluate the anesthetic effect of single-point low-volume peribulbar anesthesia supplemented by topical anesthesia. SETTING: Private ambulatory ophthalmic practice. METHODS: Five hundred consecutive patients received 4 cc of lidocaine 2% with 200 units hyaluronidase as 1-point peribulbar anesthesia. This was supplemented by lidocaine 4%, 1 drop every 3 to 5 minutes for 3 instillations. Phacoemulsification and intraocular lens implantation were performed through a scleral tunnel or clear corneal approach. All patients were evaluated for intraoperative akinesia, lid closure, and anesthesia. One hour after surgery, the eye patch was removed and patients were evaluated for pain, discomfort, foreign-body sensation, diplopia, and lid closure. RESULTS: All patients had no pain to mild discomfort during surgery; 34% had total and 58% partial akinesia; 78% had poor orbicularis action (lid closure); 12% had subconjunctival hemorrhage. Postoperatively, 42% of patients had foreign-body sensation caused by conjunctival coaptation by diathermy or corneal edema. Diplopia occurred in 32% of patients but resolved within 1 hour after eye-patch removal, and partial ptosis occurred in 58%, resolving within 2 hours of patch removal. All patients had normal lid closure when the eye patch was removed. CONCLUSIONS: Low-volume 1-point peribulbar anesthesia supplemented by topical anesthesia was safe and effective and provided early visual recovery. Topical therapy can be started 1 hour postoperatively. PMID- 9850910 TI - Alzheimer's disease: the size of the problem, clinical manifestation and heterogeneity. AB - Age is the major demographic risk factor for the development of Alzheimer's disease (AD) and the ageing of our society has contributed to the growing importance of AD. The problem of AD has been tackled from numerous clinical and scientific angles. The traditional clinico-pathological disease concept, which relies on the manifestation of a dementia syndrome and the elimination of other relevant systemic and brain diseases for a diagnosis of AD, accommodates a number of genetically and clinically heterogeneous conditions, but cannot promote our understanding of the preclinical pathogenetic processes and their interaction. If new molecular biological discoveries cannot be integrated with this traditional clinico-pathological model, it may have outlived its usefulness. PMID- 9850911 TI - Progression of Alzheimer's disease: variability and consistency: ontogenic models, their applicability and relevance. AB - Much has been learned about the clinical symptomatology of Alzheimer's disease (AD) and ontogenic reciprocal relationships in the past few decades. It is now possible to describe and verify inexorable symptomatic sequences and corresponding temporal relationships. It is also possible to identify more variable symptoms in AD. Ontogenic models can be useful in providing a clearer understanding of the nature of AD symptomatology in terms of both consistency and variability. These models can also be informative in explicating the management needs of AD patients and the treatment possibilities of AD symptoms as well as the etiology of variability in AD symptoms. PMID- 9850912 TI - Mild cognitive impairment--an early stage of Alzheimer's disease? AB - The hypothesis that mild cognitive impairment (MCI) represents an early stage of Alzheimer's disease (AD) was investigated by reviewing recent research from three sources: asymptomatic and symptomatic individuals carrying mutations that cause AD, hospital-samples of non-demented patients with MCI at the initial examination that are followed longitudinally, and community-based incident cases of AD. Studies with asymptomatic mutation carriers of the amyloid precursor protein and presenilin 1 gene have shown a linear and disease-related decline in most cognitive functions that begins approximately 10 years before the expected clinical onset of AD. However, there is considerable overlap between the level of impairment for mutation carriers and non-carriers of the same age during the early preclinical stage of AD. Hospital-based longitudinal studies have shown that non-demented individuals with isolated mild episodic memory impairment may develop clinically diagnosed AD with widespread cognitive deficits in a few years time. Community-based epidemiological studies on the incidence of AD demonstrate that indices of episodic memory, in addition to measures of general cognitive functioning, are useful in predicting early AD. In contrast, subjective memory impairment or age-associated memory impairment are less powerful predictors of future dementia development. In summary, there is converging evidence to demonstrate that preclinical AD is characterized by a common behavioral phenotype, with cognitive decline in several domains, predominantly in episodic memory. The decline appears to start many years before the clinical onset of AD. Moreover, the progression of the impairment appears to be continuous. Finally, this pattern of performance generalizes across etiology of AD (familial or sporadic), clinical onset (early or late), sample composition (hospital or community), and method of assessment. PMID- 9850913 TI - The prognosis of mild cognitive impairment in the elderly. AB - PURPOSE: To determine whether or not subtypes of intellectual functioning are suitable to predict further cognitive decline in individuals with mild cognitive impairment. DESIGN: Naturalistic longitudinal study (mean interval 2.7 years). PATIENTS: 41 subjects with mild cognitive impairment who attended a memory clinic. METHODS: SIDAM, CT, SPECT, and ApoE genotype. RESULTS: At follow-up, 8 out of 41 patients (19.5%) with MCI had progressed to dementia, 8 patients (19.5%) had improved to normal levels of cognitive functioning, 25 patients (61%) had remained stable within the MCI group. At baseline the two prognostic groups differed significantly with regard to age, memory functions, orientation, and the degree of atrophy of the left medial temporal lobe on CT scan. CONCLUSION: The majority of MCI patients in this study remained cognitively stable within the observation period. Patients with older age, poorer test performance on memory tasks and orientation deficits are at higher risk of progressive decline to dementia. CT measures of medial temporal lobe atrophy may be a sensitive parameter of group discrimination. PMID- 9850915 TI - Increase of optical illusion in demented patients. AB - Assuming a particular psychological function of optical gestalt perception, its impairment would lead to a decreasing extent of gestalt related optical illusion. An increase of optical illusion would be expected in the case of a loss of adaptability and cognitive compensation, usually revising the phenomenon of optical illusion. 16 demented out-patients were compared to 16 hospitalized schizophrenics by a 'Computerized Assessment of Change in Optical Illusion' (CACOI), measuring the extent of optical illusion by patient's assessment of 12 variations of the figure of Mueller-Lyer, differing in baseline length. The results showed a significant increase of optical illusion in demented patients compared to the schizophrenic controls (p = 0.019). Taking into account that the extent of optical illusion by the figure of Mueller-Lyer usually is decreasing with age and was found to be increased in schizophrenics, our results support the hypothesis of an early loss of adaptability and cognitive compensation in dementia. PMID- 9850914 TI - Prevalence of mild cognitive impairment in an elderly community sample. AB - The term "mild cognitive impairment" refers to cognitive deficits which exceed normal physiological aging processes, but do not fulfill the criteria for dementia. While recent studies indicate that the respective deficits can be reliably assessed, different diagnostic criteria have prevented a wide application of this diagnosis in clinical practice. The aims of the present study were (1) to assess the prevalence rates of four current diagnostic concepts and (2) to investigate mild cognitive impairment with respect to psychological and sociodemographic variables. Data from 202 probands recruited from the interdisciplinary longitudinal study on adult development were analyzed. On the time of examination, probands were between 60 to 64 years old and in a good health. The following prevalence rates were determined: 13.5% for age-associated memory impairment (AAMI), 6.5% for age-consistent memory impairment (ACMI), 1.5% for late-life forgetfulness (LLF), and 23.5% for aging-associated cognitive decline (AACD). Complaints of cognitive deficits were significantly correlated with higher scores on depression and neuroticism scales but with none of the neuropsychological measures. Reduced performance in neuropsychological tests was associated with a lower educational level and socioeconomic status. We conclude that the prevalence rates of mild cognitive impairment are highly dependent on the diagnostic criteria applied. In this respect the self-report of cognitive decline might be a less useful criteria. Longitudinal studies are warranted to further elucidate the predictive value of these diagnostic criteria. PMID- 9850916 TI - Age and dementia effect on neuropsychological test performance in very old age- influence of risk factors for dementia. AB - In old age a large part of the variance in cognitive performance in population samples is explained by normal aging; in addition many subjects over 80 years are demented and therefore dementia also explains a part of cognitive variability. The question is whether the different factors for dementia (such as ApoE4, external atrophy parameter of the cranial computer tomography [cCT], education, sex or serum zinc level) influence the relation between age or dementia and Mini Mental State (MMSE) performance. In an epidemiological study data were analyzed of N = 239 subjects for the above factors. Most statistically significant variables of the MMSE do not change the amount of the partial correlation coefficient between the parameters age or dementia and MMSE. The external atrophy, however, diminishes the magnitude of the partial correlation between age and MMSE. In contrast the dementia-MMSE relation is unchanged. This points to a generally similar factor structure of cognitive aging and dementia in old age, but differences exist with respect to the importance of the external atrophy parameter of the brain. Most factors investigated explain separate parts of variance of cognitive performance in old age. PMID- 9850917 TI - Neuropathology of Alzheimer's disease: a critical update. AB - The unequivocal diagnosis of Alzheimer's disease (AD) rests on histopathological evidence at brain autopsy or biopsy. The morphology of AD includes cerebral atrophy, deposition of beta A4 amyloid (A beta) (senile plaques and amyloid angiopathy), neuritic changes (neuritic plaques, neurofibrillary tangles (NFT) and neuropil threads) with formation of paired helical filaments (PHF) containing polymerized hyperphosphorylated tau protein triplet, causing disruption of the neuronal cytoskeleton with loss of synapses and neurons, with altered cortico cortical connectivity, leading to disconnection of the cerebral cortex. Defining criteria for the morphologic diagnosis of AD is difficult due to the phenotypic heterogeneity of the disease, the absence of specific markers, and overlap of AD morphology with that observed in non-demented elderly individuals. This gray zone between normal to pathologic aging and full-fledged AD represents an important diagnostic problem and should be overcome by better standardized criteria that will allow to minimize interrater and interlaboratory variability in the diagnosis of AD. Current criteria for the morphologic diagnosis of AD are based on (semi)quantitative assessment of diffuse and neuritic plaques (NIA), exclusively neuritic plaques (CERAD), plaques and NFT in neocortex and hippocampus (Tierney et al., 1988), and staging of hierarchic spreading of neuritic AD changes (Braak and Braak, 1991); all of them have weaknesses and need to be revalidated. Multivariant analysis of an autopsy series of elderly subjects revealed significant correlations between psychostatus and both the CERAD criteria and Braak staging. Recent recommendations of the NIA-Reagan Institute for the morphologic diagnosis of AD are presented. Although the role of plaques and NFT in the pathogenesis of AD remains undetermined, clinicopathological correlative studies have shown that both lesions, if present in sufficient numbers, particularly in the neocortex, are considered the best morphological signposts for AD. Recent studies on neuron death in AD that, at least in part, appears different from classical apoptosis and may precede the symptomatic stage of AD, have shown varying results indicating only indirect relationship between DNA fragmentation and both A beta deposition and NFTs. Both these AD-typical markers appear to increase the risk of cells to degenerate, but are not the sole responsibles of the degenerative process in AD, the basic mechanisms of which remain to be elucidated. PMID- 9850918 TI - Evolution of Alzheimer's disease related cortical lesions. AB - Alzheimer's disease is an immutably progressing dementing disorder. Its major pathologic hallmark is the gradual development of neurofibrillary changes in a few susceptible nerve cell types. The cortical changes do not occur inevitably with advancing age. Once the disease has begun, spontaneous recovery or remissions are not observed. The initial changes develop in poorly myelinated areas of the temporal lobe. The destructive process then follows a predictable pattern as it extends into other cortical areas. Advanced age is not a prerequisite for the evolution of the lesions. Alzheimer's disease is thus an age related, but not an age-dependent disease. The spread of the neurofibrillary changes resembles the process of cortical myelination, however in reverse order. PMID- 9850919 TI - What are the relations between Lewy body disease and AD? AB - Several hospital based autopsy series indicate dementia with Lewy bodies (DLB) to be the second most common pathological subtype of degenerative dementia in elderly subjects. The majority of DLB cases have high densities of beta amyloid senile plaques, whereas neocortical neurofibrillary tangle density is only slightly increased above age-matched normal control values and over tenfold lower than the average in Alzheimer's disease. The interpretation of this Alzheimer type pathology is problematic, reflecting in part changing views about the neuropathological diagnosis of AD itself. AD is characterised by hyperphosphorylation of the microtubular associated protein tau, and DLB by neurofilament abnormalities including phosphorylation, ubiquitination, proteolysis, and cross-linking of constituent proteins. The two diseases appear therefore to be distinct at an ultrastructural and molecular level, a conclusion which is consistent with the fact that the clinical syndromes associated with DLB and AD are sufficiently differentiated to allow for accurate antemortem diagnosis. PMID- 9850920 TI - The influence of coincidental vascular pathology on symptomatology and course of Alzheimer's disease. AB - The aim of this review is to determine the influence of coincidental cerebrovascular pathology on the symptomatology and course of Alzheimer's disease (AD). The link between stroke and AD is probably higher than expected by chance for the following reasons: (i) both pathologies share genetic risk factors such as the epsilon 4 allele of the apolipoprotein E gene; (ii) AD patients have changes in the brain vessels that may lead to either ischemic or hemorrhagic stroke or white matter changes or both; (iii) there is evidence of an increased risk of stroke in AD patients; (iv) there is evidence of a frequent association of AD and stroke at autopsy. Because of the summation of the various types of lesions, stroke lesions may lead to an increase progression of cognitive decline in AD patients. Recognition of a vascular component in a dementia syndrome is therefore useful for the management of AD patients. Whether an optimal management of risk factors for stroke may delay the clinical expression of dementia in patients with preclinical Alzheimer pathology should be evaluated. PMID- 9850921 TI - Corpus callosum in Alzheimer's disease and vascular dementia--a quantitative magnetic resonance study. AB - We investigated atrophic alterations in different regions of the corpus callosum in Alzheimer's disease (AD) and vascular dementia (VD) with respect to clinical changes. 32 patients with AD (NINCDS-ADRDA criteria), 17 patients with VD (NINDS AIREN criteria) and 13 healthy control subjects were included. 3-D MRI sequences were acquired using a 1.5T MRI scanner. The size of the corpus callosum and its subdivisions was sampled on 5 mid-saggital slices using a personal computer-based software. Total callosal size was significantly reduced in AD but not in VD. Furthermore, the most rostral parts of the corpus callosum were significantly smaller in AD when compared to controls. Again, these changes were not found in patients with VD. Severity of dementia was significantly correlated with the size of the midbody of the corpus callosum in AD. Callosal atrophy in AD may reflect the severity and pattern of cortical neuronal damage occurring mostly in the inferior frontal, anterior parietal and midtemporal regions. Correlations between regional callosal atrophy and severity of dementia indicate that interhemispheric cortico-cortical disconnections may contribute to the dementia syndrome. PMID- 9850922 TI - What do we learn from a few familial Alzheimer's disease cases? AB - Alzheimer's disease is the most common form of dementia. About 90% of the cases occur sporadically whereas in 10% of the cases mutations were found within three different genes. Mutations in the gene encoding the beta-Amyloid precursor protein (beta APP) are located in the ultimate neighborhood of the three proteases (secretases) involved in proteolytic processing of beta APP. These mutations cause an increased production of the long form of Amyloid beta-peptide (A beta) the major component of Amyloid plaques. In contrast to the 40 amino acid form (A beta 40), the 42 amino acid form (A beta 42) aggregates more rapidly, kills cultured neurons more efficiently, and precipitates preferentially in amyloid plaques. Interestingly, mutations in the Presenilin genes which are responsible for more then 40% of all familial AD cases also cause enhanced production of the elongated form of A beta. Therefore mutations in three different genes directly effect A beta production in a pathological manner, which strongly supports the amyloid cascade hypothesis. PMID- 9850923 TI - Aberrancies in signal transduction and cell cycle related events in Alzheimer's disease. AB - Neurodegeneration in Alzheimer's disease (AD) is associated with the appearance of dystrophic neuronal growth profiles that most likely reflects an aberrant attempt of neuronal repair. This process of neuronal reorganisation, which eventually goes awry and becomes a disease itself, might be initiated physiologically as a response to neuronal injuries. Minor neuronal damage due to a variety of life events or genetic pertubations that are usually compensated in the normal adult brain by adaptation and repair might thus be amplified and accumulated, thereby resulting in a progressive neurodegeneration. The present paper summarizes recent evidence supporting the hypothesis that a primary impairment of intracellular signal transduction that is mediated by a hierarchy of phosphorylation signals and associated with a aborted attempt of neurons to re enter the cell-cycle is a key element in the pathomechanism of AD. These changes might result in malfunction of neuronal adaptation and repair and eventually lead to neuronal death. During the process of aging as well as in chronic neurodegenerative disorders such as Alzheimer's disease (AD), the continuous but rather slow action of pathogenetic factors might give room for the activation of compensatory mechanisms, serving to regain a neuronal population much of its synaptic connectivity in the presence of cell loss. An upregulation in the capacity to synthesize and store neurotransmitters (Lapchak et al., 1991), an increased expression of trophic factors (Hellweg et al., 1990; Arendt et al., 1995a,b), as well as regenerative sprouting (Fritschy and Grzanna, 1992), synaptic enlargement (Scheff et al., 1990; Lippa et al., 1992), and neosynaptogenesis (Ramirez and Ulfhake, 1992) have been described among other processes. PMID- 9850924 TI - Mechanisms of cell death in Alzheimer disease--immunopathology. AB - Lesions in such chronic neurodegenerative disorders as Alzheimer disease, Parkinson disease, the parkinsonism dementia complex of Guam and amyotrophic lateral sclerosis have associated with them a variety of proteins known to be involved in inflammatory processes. This is particularly true of Alzheimer disease where inflammatory reactions are thought to be important contributors to the neuronal loss. They include complement proteins, complement inhibitors, acute phase reactants, inflammatory cytokines, proteases and protease inhibitors. Studies of cultured human astrocytes and microglia, obtained from postmortem brain, have established that nearly all of these proteins are produced by one or another of these cell types. Human neurons also produce many inflammatory proteins and their inhibitors, creating complex interactions. Accumulations of amyloid and extracellular tangles apparently act as irritants, causing the activation of complement, the initiation of reactive changes in microglia, and the release of potentially neurotoxic products. Such products include the membrane attack complex, oxygen free radicals and excess glutamate. Twenty epidemiological studies that have been published to data indicate that populations taking antiinflammatory drugs have a significantly reduced prevalence of Alzheimer disease or a slower mental decline. One small clinical trial with indomethacin showed arrest of the disease over a 6 month period. Therapeutic intervention in key inflammatory processes holds great promise for the amelioration of Alzheimer disease and possibly other neurodegenerative disorders. PMID- 9850925 TI - Pathological immuno-reactions of glial cells in Alzheimer's disease and possible sites of interference. AB - A significant role of a pathological glial cell activation in the pathogenesis of Alzheimer's disease is supported by the growing evidence that inflammatory proteins, which are produced by reactive astrocytes, promote the transformation of diffuse beta-amyloid deposits into the filamentous, neurotoxic form. A number of vicious circles, driven by the release of TNF-a and free oxygen radicals from microglial cells, may cause an upregulated microglial activation and their production of interleukin-1 which triggers, secondarily, the crucial activation of astrocytes. Reactive functional changes of glial cells seem to be controlled by an altered balance of the second messengers Ca2+ and cAMP and can be counterregulated by the endogenous cell modulator adenosine which strengthens the cAMP-dependent signalling chain. A further reinforcement of the homeostatic adenosine effects on glial cells by pharmaca, such as propentofylline, may add to neuroprotection in Alzheimer's disease. PMID- 9850926 TI - Therapeutic strategies based on immunological hypotheses of Alzheimer's disease. AB - There are several findings demonstrating the importance of inflammatory or immunological processes in the etiopathogenesis of Alzheimer's disease (AD). Several studies on nonsteroidal anti-inflammatory drugs (NSAID) give evidence for a therapeutic effect of these drugs in AD, among other the prospective double blind placebo-controlled study on indomethacin, strongly support the therapeutic effect of these agents. Possibly also the therapeutic effects of propentofylline might be interpreted in the light of this theory. PMID- 9850927 TI - Risk factors for Alzheimer's disease during aging. Impacts of glucose/energy metabolism. AB - The majority of Alzheimer patients is of late onset and with unknown etiology. However, several risk factors have been discussed among which age is a most important one with respect to sporadic Alzheimer type dementia (SDAT). Age includes changes in brain glucose/energy metabolism, in both insulin and acetylcholine signal transduction and in membrane function to name the functionally most important ones. Variations in these parameters can form the basis for ongoing changes in terms of the principle of self-organized critically inducing catastrophic i.e. disease processes. Subsequent abnormalities at the cellular and molecular levels may develop including the formation of both amyloid plaques and neurofibrillary tangles. PMID- 9850928 TI - Aging, dementia and calcium metabolism. AB - After a brief discussion of the theory of disturbed neuronal calcium metabolism an overview is given on the results of clinical studies with nimodipine in the treatment of vascular and Alzheimer's dementia. PMID- 9850929 TI - Changes of intracellular calcium regulation in Alzheimer's disease and vascular dementia. AB - Free intracellular calcium ([Ca2+]i) represents probably the most important intracellular messenger for many signal transduction pathways. Due to this crucial role of [Ca2+]i, it has been assumed that alterations of [Ca2+]i are critically involved in brain aging and in the pathophysiology of Alzheimer's disease (AD). This hypothesis is corroborated by several studies demonstrating changes of [Ca2+]i in peripheral cells from AD patients. However, the findings are still controversial. Using blood lymphocytes and neutrophils as two different peripheral model systems, we evaluated several parameters of intracellular Ca2+ regulation in a very large group of AD patients and non-demented controls. We found no major difference in Ca2+ homeostasis, since neither the basal [Ca2+]i, nor the activation-induced Ca2+ responses differed among neutrophils or lymphocytes from aged controls and AD patients. However, we observed a delayed Ca2+ response of AD lymphocytes after phytohemagglutinin (PHA) stimulation indicating an impaired function of Ca2+ influx-controlling mechanisms. Furthermore, we studied whether differences exist in Ca2+ regulation between lymphocytes from patients with vascular dementia and AD patients, to define AD specific alterations and to distinguish between the two dementia groups and non demented control subjects respectively. First evidences indicate that Ca2+ mobilization in lymphocytes is specifically impaired in lymphocytes from patients with vascular dementia. PMID- 9850930 TI - Free radicals in Alzheimer's dementia: currently available therapeutic strategies. AB - Substantial evidence now exists that oxidative stress may play an important role in the etiopathogenesis of DAT. The different sources of oxidative stress in DAT are suggesting several pharmacological opportunities for influencing the disease. It is possible to distinguish 2 major types of possible therapeutic agents according to their pharmacological point of attack. 1. Radical scavengers, agents directly interacting with free radicals. Candidates of this type are gingko biloba, vitamins A, C, E and estrogen. 2. Antioxidants, which are able to prevent or decrease the production of free radicals by use of specific neuropharmacological properties. Candidates are selegiline, a MAO-B inhibitor well established in the therapy of Parkinson's disease, and tenilsetam, which is believed to be an AGE-inhibitor. Recent in vitro studies have demonstrated the efficacy of both types of therapeutic agents by preventing or delaying oxidative neural damage. Some clinical data exist regarding the antidementive properties particularly in terms of gingko biloba, selegiline and vitamin E. The efficacy studies about these compounds seem to indicate a promising future strategy in the therapy of DAT. But it is too early to draw definite conclusions since it is well known that all of our candidate substances do not act specifically as radical scavengers or antioxidants. PMID- 9850931 TI - Free radicals in Alzheimer's disease. AB - Alzheimer's disease is a neurodegenerative disorder comprising multisystem atrophies probably caused by multifactorial processes. The disease is characterized by typical neuropathology, impaired synaptic function and massive cell loss. The pathobiochemistry of this disorder involves oxidative stress, which accumulates free radicals leading to excessive lipid peroxidation and neuronal degeneration in certain brain regions. Moreover, radical induced disturbances of DNA, proteins and lipid membranes have been measured. The hypothesis has been proposed that cellular events involving oxidative stress may be one basic pathway leading to neurodegeneration in Alzheimer's disease. In this work we report evidence for increased oxidative stress and disturbed defense mechanisms in Alzheimer's disease, which may result in a self-propagating cascade of neurodegenerative events. Furthermore it is evident from experimental data, that aggregation of beta-amyloid and beta-amyloid toxicity is favourably caused by oxidative stress. Therefore, oxidative stress plays a key role in the conversion of soluble to unsoluble beta-amyloid, suggesting that oxidative stress is primary to the beta-amyloid cascade. PMID- 9850932 TI - Simulation of cortical cholinergic deficits--a novel experimental approach to study pathogenetic aspects of Alzheimer's disease. AB - Cholinergic lesion paradigms have been used to study the role of the cholinergic system in cognitive function, and its implication in cognitive deficits that occur in Alzheimer's disease. In the last few years an increasing number of studies have applied neurotoxins including excitotoxins or cholinotoxins to produce reductions in cortical cholinergic activity. One of the most serious limitations of these lesion paradigms is the fact that the cytotoxins used are far from being selective to cholinergic cells. Recently, a monoclonal antibody to the low-affinity nerve growth factor (NGF) receptor, 192IgG, coupled to a cytotoxin, saporin, has been described as an efficient and selective immunotoxin for the NGF-receptor bearing cholinergic neurons in rat basal forebrain. Here we demonstrate the usefulness of 192IgG-saporin as a powerful tool for producing an animal model with selective and specific basal forebrain cholinergic lesions in rats which can be applied to simulate some neurochemical sequelae of Alzheimer's disease including cholinergic mechanisms in processing of the amyloid precursor protein, and could be of particular value to elaborate and to test therapeutical strategies compensating for the reduced cortical cholinergic input. PMID- 9850933 TI - Neuronal anion exchange proteins in Alzheimer's disease pathology. AB - Anion exchange (AE) proteins are present in human neurons in the brain. Immunohistochemical data indicate that their apparent expression level increases with age, and especially with degeneration in Alzheimer's disease-affected brain areas. The increase in immunoreactivity is probably caused by changes in AE structure that lead to an increased accessibility of hitherto hidden epitopes. These epitopes correspond to regions in the membrane domain that are involved in generation of senescent cell-specific antigen from AE1 in aging erythrocytes. Elucidation of the molecular nature of these changes and the underlying mechanisms will lead to insight in the processes that govern aging- and degeneration-associated perturbation of membrane integrity. The functional consequences of changes in AE structure may range from acidosis and disturbance of cytoskeleton integrity to untimely or impaired recognition of neurons by microglia. PMID- 9850934 TI - Lymphocytes as cell model to study apoptosis in Alzheimer's disease: vulnerability to programmed cell death appears to be altered. AB - Recent evidence indicates that programmed cell death (apoptosis) may contribute to neuronal death in Alzheimer's disease (AD). In situ data derived from post mortem brain tissue indicate that DNA fragmentation which represents an important and typical apoptotic feature is markedly increased in brain cells of AD patients compared to controls. Furthermore, in vitro studies demonstrate that the peptide beta-amyloid (A beta) and its fragments induce apoptosis in neuronal cell cultures. One possible mechanism initiating apoptosis could be free radical generation by the peptide leading to oxidative stress. In a wide range of cell types common morphological and molecular events occur during apoptosis and several genes appear to be involved. Particularly in lymphocytes, apoptosis plays an important physiological role. Our data demonstrate that similar oxidative stressors induce apoptosis in mature human lymphocytes as in neurons. In addition, first evidence indicates that susceptibility to apoptosis is altered in lymphocytes from AD patients compared to non-demented controls. Our preliminary findings suggest that changes of the individual sensitivity to undergo cellular apoptosis are already detectable in lymphocytes from AD patients, probably as a consequence of genetic as well as other risk factors. Therefore, this biochemical marker might have the potential for identifying individuals at risk of the diseases. PMID- 9850935 TI - Potential link between interleukin-6 and arachidonic acid metabolism in Alzheimer's disease. AB - Prostaglandins (PGs) and cytokines, such as interleukin-1 (IL-1) and interleukin 6 (IL-6), have been implicated in the etiopathology of various inflammatory and degenerative disorders, including Alzheimer's disease (AD). Previously, we detected the presence of IL-6 in cortices of AD patients. On the other hand, non steroidal antiinflammatory drugs (NSAIDs), potent inhibitors of prostaglandin synthesis, have been shown to be beneficial in the treatment of AD. Until now, it remained unclear whether and how these two observations were functionally connected. Here, we show that PGs are able to induce IL-6 synthesis in a human astrocytoma cell line. PGE1 and PGE2, but not PGD2 and PGF2 alpha, led to a rapid and transient induction of astrocytic IL-6 mRNA, followed by IL-6 protein synthesis. Furthermore, PGE2 potentiated IL-1 beta-induced IL-6 mRNA synthesis. These results suggest a possible link between the release of PGs from activated microglia and the astrocytic synthesis of IL-6, which itself may affect neuronal cells, as hypothesized for Alzheimer's disease. Finally we demonstrate that microglia are a strong source of PGE2 synthesis indicating that these cells may act as the origin of the pathogenic cascade. PMID- 9850936 TI - Neurotrophin binding to the p75 neurotrophin receptor is necessary but not sufficient to mediate NGF-effects on APP secretion in PC-12 cells. AB - In the present study the pheochromocytoma cell line (PC-12) was used as a model system to determine the role of the two neurotrophin receptors in the regulation of amyloid precursor protein (APP) secretion by nerve growth factor (NGF). To stimulate TrkA and/or p75NTR signaling in PC-12 cells, we used NGF, brain-derived neurotrophic factor (BDNF), and NGF in the presence of an excess of BDNF or the monoclonal antibody 192IgG, to block p75NTR binding to NGF. Our results demonstrate that NGF stimulates APP secretion in a dose dependent fashion with maximum effects at 10 ng/ml, known to saturate high-affinity NGF binding sites. Treatment of PC-12 cells with varying concentrations of BDNF, 1-1,000 ng/ml, did not alter APP secretion, suggesting that binding to p75NTR alone is not sufficient to affect APP secretion. When blocking NGF binding to p75NTR with BDNF or 192IgG, on the other hand, NGF effects on APP secretion were abolished. These findings suggest that in cells expressing p75NTR and TrkA receptors, binding of NGF to the p75NTR is required to mediate NGF effects on APP secretion. Our data are also consistent with a proposed function of the p75NTR in receptor recruitment and "presentation" of NGF to receptors. PMID- 9850937 TI - Aromatic alcohols as neuroprotectants. AB - Free radicals and oxidative stress-induced neuronal cell death have been implicated in various neurological disorders including neurodegenerative diseases. We have recently shown that estrogens can protect neurons against oxidative stress due to their antioxidant potential. Here, we report that also other aromatic alcohols with intact phenolic groups and different phenol derivatives can protect neurons against oxidative cell death as induced by glutamate and hydrogen peroxide. Starting with melatonin, in this study, we degraded the chemical structure step by step and tested various aromatic alcohols as well as phenol derivatives for their potential antioxidant activity. We found that aromatic alcohols with intact phenolic groups protect neurons effectively against oxidative damage and cell death and that this neuroprotective activity is independent of the time the compound is added before the toxin. Methylation of the phenolic hydroxyl group led to a decrease or loss in neuroprotection. Moreover, the tested compounds directly inhibited peroxidation reactions suggesting that the neuroprotection is mediated by antioxidant properties. Our result may have some impact on the design of future drugs for the antioxidant treatment or prevention of oxidative stress-associated pathological conditions. PMID- 9850938 TI - The therapeutic potential of tacrine. AB - Tacrine, a non-competitive inhibitor of acetylcholine esterase, has been approved for the treatment of patients with mild to moderate Alzheimer's disease in 1995. The available evidence suggests that in a substantial proportion of patients tacrine has the potential to improve cognitive functions, to ameliorate behavioural problems, and to delay the time to significant clinical endpoints. Thus, tacrine meets realistic expectations from symptomatic treatment. The major safety concern about tacrine is the probability of liver enzyme elevations and the high frequency of adverse events, particularly gastrointestinal. Findings are presented from an open-label multicentre study showing that tacrine is a safe and manageable compound in private practice. PMID- 9850939 TI - Sustained efficacy and safety of idebenone in the treatment of Alzheimer's disease: update on a 2-year double-blind multicentre study. AB - The 2-year efficacy and safety of idebenone were studied in a prospective, randomized, double-blind multicentre study in 3 parallel groups of patients with dementia of the Alzheimer type (DAT) of mild to moderate degree. A total of 450 patients were randomized to either placebo for 12 months, followed by idebenone 90 mg tid for another 12 months (n = 153) or idebenone 90 mg tid for 24 months (n = 148) or 120 mg tid for 24 months (n = 149). The primary outcome measure was the total score of the Alzheimer's Disease Assessment Scale (ADAS-Total) at month 6. Secondary outcome measures were the ADAS cognitive (ADAS-Cog) and noncognitive score (ADAS-Noncog), the clinical global response (CGI-Improvement), the SKT neuropsychological test battery, and the Nurses' Observation Scale for Geriatric Patients (NOSGER-Total and IADL subscale). Safety parameters were adverse events, vital signs, ECG and clinical laboratory parameters. During the placebo controlled period (the first year of treatment), idebenone showed statistically significant dose-dependent improvement in the primary efficacy variable ADAS Total and in all the secondary efficacy variables. There was no evidence for a loss of efficacy during the second year of treatment, as a further improvement of most efficacy variables was found in the second year in comparison to the results at the 12 months visit. Also, a clear dose effect relationship (placebo/90 mg < idebenone 90 mg < idebenone 120 mg) was maintained throughout the second year of treatment. This suggests that idebenone exerts its beneficial therapeutic effects on the course of the disease by slowing down its progression. Safety and tolerability of idebenone were good and similar to placebo during the first year of treatment and did not change during the second year. PMID- 9850940 TI - Clinical experience with Donepezil (Aricept) in the UK. AB - Experience of the use of Donepezil (Aricept) in the UK since licensing is discussed. The results of a 30 week double blind parallel group study in the US of Donepezil 5 or 10 mg versus placebo show statistically significant improvements in cognitive and clinical global assessments. Beneficial effects were demonstrated in the absence of significant adverse effects on physical or laboratory values. The guidelines for use adopted in the UK are discussed with the emphasis on identifying non-cognitive behaviours, as significant improvements in these have been found and often have more impact on quality of life than cognitive improvements alone. PMID- 9850941 TI - Revisiting restraints. PMID- 9850942 TI - Tattoos and piercing offer psychopathology clues. PMID- 9850943 TI - Sexual predators do not always fit the "profile". PMID- 9850944 TI - Consider parents when examining suicide risk in kids. PMID- 9850945 TI - Battling back from childhood sexual abuse and surviving the journey. AB - The experience of childhood sexual abuse can be so profound that it fundamentally alters one's development and personality. Avoidance, repression, denial, and rationalization are some defense mechanisms used by the victims to deal with the abuse. Nurses working with a patient with a history of childhood sexual abuse must help the patient discover his or her own personal meaning of the abuse. PMID- 9850946 TI - Current understanding of violence and aggression: assessment and treatment. AB - Links between violence, aggression, and mental illness are well documented. Despite this association, our current understanding of the causation and optimal treatment of aggression remains limited. This lack of knowledge is alarming because nurses treating patients with mentally illness are frequent targets of patient aggression. Consequently, the aim of this article is to provide contemporary information regarding the concept of patient aggression, assessment of violent behavior, and implementation of treatment interventions. A review of pharmacological and psychosocial strategies are presented as well. These findings provide psychiatric nurses with a conceptual model as well as practical interventions for patient aggression. PMID- 9850947 TI - Prisoners as patients. The experience of delivering mental health nursing care in an Australian prison. AB - Correctional or prison mental health nursing is a highly specialized area of practice that has undergone substantive role development in recent years. However, little research has explored aspects of prison-based nursing practice or practice arrangements. The experience of delivering mental health nursing care in prison can be disempowering, resulting in feelings of frustration, isolation, and stigma. In developed nations, prison mental health nurses face the rewarding challenge of gaining greater recognition for the specialized nature of their practice and their key role in the correctional and criminal justice continuum. PMID- 9850948 TI - Levels of recovery from psychotic disorders chart. AB - The Levels of Recovery From Psychotic Disorders Chart was developed by this author at the request of the Massachusetts Alliance for the Mentally Ill (AMI). This chart is used to assess and treat patients' recovery from psychotic illnesses. By using the chart in a process of periodic assessment, patients, their families, and clinicians are prompted to consider alternative antipsychotic medications that might potentiate recovery. This article describes the development of the chart, methods of recording observations, clinical recommendations based on the chart, and implications for nursing. PMID- 9850949 TI - Electrophysiological evidence of a relationship between idiopathic carpal and tarsal tunnel syndromes. AB - Electrophysiological findings are reported suggesting a relationship between idiopathic tarsal (TTS) and carpal tunnel syndromes (CTS) to explain that the coexistence of both entrapment syndromes in the same patients is not coincidental. Sixty-five patients with idiopathic CTS and 15 with idiopathic TTS were selected. None of the patients with CTS reported any symptoms, nor did they have any signs of TTS, and vice versa. Distal sensory conduction velocity (SCV) of the tibial nerve was reduced in ten of 65 patients with CTS; in five of these ten patients, tibial distal motor latency (DML) was also delayed. Reduced SCV and increased DML were evident in the median nerve of two patients with TTS. The mean of DML and SCV of the tibial nerve in the CTS group and of the median nerve in the TTS group were significantly reduced with respect to the control group without differences in the conduction of the ulnar, deep peroneal and sural nerves. This indicates that there was subclinical involvement of the median and tibial nerves in these patients. Besides having a narrow carpal tunnel, patients with CTS presumably also have a narrow tarsal tunnel and vice versa, and are therefore prone to develop both nerve entrapment syndromes. The different incidence of the two syndromes is explained on the basis of anatomical and functional differences between the two nerves. PMID- 9850950 TI - Ageing effects on flash visual evoked potentials (FVEP) recorded from parietal and occipital electrodes. AB - The effects of ageing on flash visual evoked potentials (FVEP) recorded from 6 posterior parietal and occipital sites were studied in a sample of 73 healthy subjects of between 20 and 86 years of age. Latencies of components P1, N1 and P2, and amplitudes of components P1 and P3 increased linearly with age at all emplacements. The results obtained from occipital electrodes are in line with previous reports and additionally show that i) the effects of age constantly increase over time, and ii) age affects not only the early but also the later components (> 150 ms) of the FVEP. The overall pattern of results suggests that elderly subjects show slower transmission of visual information and deficiencies in the inhibitory regulation of activity generated during the arrival of repetitive non-attended visual stimulation. The findings with parietal electrodes show that ageing effects are more marked at these emplacements than at occipital electrodes. Furthermore, this raises the question of a possible differential involvement of primary and nonprimary visual cortex by age, but this hypothesis can only be explored with high-intensity multichannel recordings and dipolar modelling. PMID- 9850951 TI - Low intensity galvanic vestibulo-ocular reflex in normal subjects. AB - An electrical stimulation in man applied between the two mastoids could facilitate the distinction between labyrinthine and retrolabyrinthine lesions by stimulating directly the primary vestibular afferences. However, for this test to be really effective in current medical practice, the results obtained in normal subjects must be symmetrical and reproducible one day to another. The ocular responses induced by a constant electrical stimulation of 2.5 mA, applied between the two mastoids for 30 s (electrically evoked vestibulo-ocular reflex [EVOR]), in one direction and the other, were quantified in ten healthy subjects. Each subject was studied in two different sessions separated by 1 week. Horizontal eye movements were recorded in darkness by an infrared light reflection eye-tracking system. Slow-phase velocity and nystagmus frequency were about 20% higher when the cathode was on the right mastoid than when it was on the left mastoid. This directional preponderance (DP) displayed large individual differences between the two sessions. The reproducibility of the reflectivity (mean of right and left EVOR) was high (r about 0.8). The weak reproducibility of the DP makes the EVOR at weak intensity inadequate to evaluate unilateral vestibular hypofunction. On the other hand, because of the high reproducibility of reflectivity, the EVOR should be effective in detecting bilateral vestibular hypofunction. Moreover, because of the weak intensity of stimulation, no local anaesthesia is needed so the manoeuvre is easy to repeat in case of chronic diseases. PMID- 9850952 TI - [Indications for emergency EEG in perinatal pain]. AB - EEG is a basic tool in neurological evaluation of fullterm or premature newborn. An emergency recording (in hours following birth) is required in two circumstances: a suspicion of seizure (especially before starting specific therapy), and a major behavioral disorder. The main cause is, in fullterm newborn, hypoxic-ischemic encephalopathy. More precise informations are provided by recording between 10 and 48 hours after birth. In premature, emergency EEG may be recorded between 24 and 48 hours after birth. Some particular EEG features suggest specific causes such as metabolic or infectious diseases and focal lesions. In some cases, especially during seizures, conventional EEG can be completed by continuous monitoring. PMID- 9850953 TI - [[Emergency pediatric EEG in mental confusion, behavioral disorders and vigilance disorders: a retrospective study]. AB - Recording of electroencephalogram (EEG) is of value to estimate vigilance states in children as in adults. In order to determine the diagnostic and prognostic value of emergency EEG in case of mental confusion, behavioral disorders and vigilance disorders in childhood, we conducted a retrospective study in 397 children (aged 2 months to 16 years). EEG was recorded less than 24 hours after an emergency consultation for acute confusion or acute behavioral disorder (n = 106) or after admission to the intensive care unit for comatose stage (n = 291). EEG gave diagnostic indications mainly in case of convulsive pathology or hepatic encephalopathy. In comatose children, we established a 4-stage EEG scale of increasing severity. This classification was compared to EEG scales already published in the literature and appeared very similar to that from Pampiglione and Harden, established in 150 children after cardiac arrest. A highly poor prognostic value was associated with burst-suppression post-anoxic patterns and with isoelectric records signaling brain death. Our classification of emergency EEG patterns is mainly helpful in these two situations, but does not exclude strict and repeated clinical and EEG follow-up in other cases, as a relatively preserved initial EEG may later deteriorate. PMID- 9850954 TI - Clinical-neuroendocrinological syndromes due to lesions of the cingulate gyrus in humans. AB - Clinical and neuroendocrinological studies were performed in 41 patients with arteriovenous malformations (AVM) of the cingulate gyrus. Before surgery, the primary affliction in 38 patients consisted of memory disturbance, 5 of these having signs of Korsakov's syndrome. Autonomic and epileptic fits were not typical. Of 38 patients undergoing surgery, postoperative worsening of memory occurred in 23, but no qualitatively new disturbances appeared. Korsakov's syndrome appeared de novo after surgery in 3 patients. The extent of memory disturbances correlated with the extent of lesions to the cingulate gyrus and with the extent to which AVM affected the corpus callosum. Qualitative analysis of memory loss syndromes revealed alterations in trace selectivity in nearly all patients and failure to retain the meaning of stories, which was accompanied by lack of insight into the patients' abnormalities. The signs of these syndromes showed clear similarity with memory deficiencies in patients with frontal lesions. This suggests that not only the frontal lobes themselves, but also their connections, are involved in producing the clinical picture in humans. PMID- 9850955 TI - Relationships between the epiphysis and hippocampus during formation of a stress response. AB - Lesions to the dorsal hippocampus and removal of the epiphysis had differently directed effects on behavior in rats in conditions of a conflict situation and on the temporal dynamics of forced swimming. The combination of both operative procedures resulted in weakening of the behavioral changes typical of hippocampus deficiency. It is suggested that the anti-stress properties of epiphyseal factors may be mediated by changes in the functional state of the hippocampus. PMID- 9850956 TI - Effects of limiting afferent flow in early postnatal ontogenesis on the development of defensive reflexes in adult rats. PMID- 9850957 TI - Induction of the c-fos gene in the chick brain during visual imprinting. PMID- 9850958 TI - Spatial organization of electrical processes in the brain: problems and solutions. AB - Theoretical questions of the spatial organization of electrical activity in the brain are discussed in terms of a multilevel realization of the synergetic principle for formation of functional systems underlying behavior and mental function. The role of the spatial-temporal superimposition of coherent structures of biopotentials in generating fields of increased activity in the cerebral cortex is discussed, these being responsible for integrative and associative functions. A hypothesis is proposed regarding the relationship between the energy and information factors of coherent structures as one of the important characteristics describing the efficiency of energy-informational processes. Emphasis is placed on the need for considering not only linear, but also nonlinear associations of biopotentials in considerations of the form and functional sense of their spatial organization. PMID- 9850959 TI - Bursts of high-frequency synchronized electrical activity in the dog neocortex during food-related operant conditioning. AB - Bursts of high-frequency (HF, 80-90 Hz, 70-80 microV) oscillations in the electrical activity (EA, 1-200 Hz) of the dog neocortex were studied during operant conditioning. These bursts of HF oscillations appeared in the EA of interstimulus intervals at the generalization stage on a background of dominant oscillations of lower frequency and amplitude (10-40 microV). Use of a new strategy for primary analysis of EA production (specifically, a coefficient of inhomogeneity) allowed amplitude-frequency inhomogeneity of the EA to be estimated, with isolation of bursts of HF oscillations. Use of an original nonharmonic analysis, consisting of expansion of EA waves into a system of half waves which were used to construct distribution maps, revealed the regional properties of bursts of HF oscillations. The results of these investigations supplement previous data obtained using other methodological approaches (Fourier transformation and spectral density factor analysis). The properties of bursts of HF oscillations observed here provide evidence for the differential involvement of cortical areas (even close-lying areas separated by distances of 3-5 mm) in the spatial-temporal organization of potentials typical of this conditioning paradigm. PMID- 9850960 TI - Background gamma-oscillations in neuronal networks with interhemisphere connections. AB - Results obtained in studies of the high-frequency components of EEG recordings and in modeling, determining the conditions for the appearance of gamma oscillations in interneuronal interactions, were compared with features of the background gamma oscillations recorded in the activity of interacting neurons located in symmetrical loci of the right and left hemisphere motor areas in anesthetized rats. Similarities in high frequencies extracted from EEG recordings and in the most commonly observed gamma oscillation frequencies suggested that these oscillations may represent one of the mechanisms underlying the high frequency EEG component. Published modeling data indicating that the formation of these oscillations involves reciprocal inhibitory connections, along with our own data that interhemisphere oscillations are seen 1.5 times more commonly than ipsilateral oscillations, suggested that transcallosal inhibition is more effective than inhibition between neighboring cells. Simultaneously extracted background oscillations in the interacting activity of callosal cells and neighboring cells could be different, as could those characterizing the activity of individual neurons. It is suggested that these differences underlie the functional heterogeneity of local cortical neuronal networks and explain the fact that these networks contain various types of inhibitory neurons. PMID- 9850961 TI - Chaotic components in the high-frequency EEG of rabbit cortex during formation of a conditioned defensive reflex. AB - The chaotic component of the high-frequency EEG of the rabbit cortex was studied during the development of a conditioned defensive reflex to nonrhythmic light stimulation with electric reinforcement applied to the animal's paw. Regular changes in the dynamic parameters of the chaotic component of the high-frequency EEG were demonstrated. Characteristic changes in the asymptotic evaluations of the correlational dimensionality of the attractor of the chaotic component (DCC) of the high-frequency EEG were detected in the cortical regions studied both during the action of the conditioned stimulus and in background conditions before presentation of combinations. These changes affected both the afferent and efferent cortical regions of the conditioned reflex arc, as well as regions not directly addressed by the conditioned or unconditioned stimuli. These results allow the dynamics of changes in the morphofunctional structures of the conditioned reflex to be seen at the cortical level during learning. Significant differences in the early and late stages of conditioned reflex formation were detected. PMID- 9850963 TI - Role of the vagus nerves in neophobia and conditioned-reflex taste aversion. AB - Combination of ingestion of water with discomfort in rats with intact vagus nerves on selection between water and saccharine solution (an unknown taste) produced aversion not to water but to saccharine, with sharp increases in water consumption. In vagotomized rats, this combination led to a significant increase in saccharine consumption with no change in water intake. After extinction of neophobia to saccharine, combination of ingestion of water with rotation induced aversion to water in both groups (this being delayed in vagotomized rats). Vagus nerve signaling activity in selection conditions appears to determine the choice of behavior strategy. PMID- 9850964 TI - Action of modulated electromagnetic fields on the emotional component of the systems organization of behavioral acts in rats. AB - This article reviews experimental data providing evidence on the effects of modulated electromagnetic fields of 30-120 V/m with a carrier frequency of 30 MHz modulated sinusoidally at frequencies of 2-50 Hz on the emotional responses accompanying various stages in the systems organization of behavior in rats. The blocking effects of fields were demonstrated in self-stimulation models in different types of conditioned reflex behavior, as well as during extinction of conditioned reflex responses in individual conditions and emotional intercourse. It is suggested that modulated electromagnetic fields which have information effects on the body act on the information component of behavior, i.e., emotion. PMID- 9850962 TI - EEG frequency ranges during perception and mental rotation of two- and three dimensional objects. AB - Spectral EEG powers were compared in 4 frequency ranges (8-13, 15-25, 25-35, and 35-45 Hz) in a group of 20 subjects during the performance of tasks requiring mental rotation of two- and three-dimensional objects. Only those EEG segments corresponding to tasks with identical solution times were analyzed. The spectral powers of oscillations in the alpha range were higher in control conditions than during task performance. Power in the frequency range 15-45 Hz was greater during task performance than in control conditions; this supports the concept that alpha rhythm desynchronization accompanies the synchronization of higher-frequency EEG rhythms. Frequency power during task performance with two-dimensional objects was greater than that during tasks with three-dimensional objects. Since the angle of rotation between two-dimensional objects was greater than that between three dimensional objects, this factor, rather than the depth of the perceived space, increased the level of cortical activation. In all experimental situations, power at frequencies of 15-45 Hz was significantly greater in the occipital regions than any other regions, reflecting the visual modality of the stimulus. Particular changes were noted in the gamma range (35-45 Hz), where power in the first second of task performance was significantly higher than in the second second; this may provide evidence that this range is more closely associated with perception and recognition processes than with mental transformation of the image. PMID- 9850965 TI - Effects of delta-sleep-inducing peptide on NMDA-induced convulsive activity in rats. AB - Acute experiments on rats showed that the ED100 of NMDA for induction of clonic convulsions was 0.53 microgram, while the ED100 of NMDA for inducing tonic extension of the forelimbs was 5.02 micrograms/animal. Determination of these parameters after administration of delta-sleep-inducing peptide (100 micrograms/kg, i.p.) revealed 2.3- and 4.46-fold increases. These results provide evidence for a neuroprotective role of delta-sleep-inducing peptide in relation to excitatory amino acid receptor agonists. PMID- 9850966 TI - Relationship between the characteristics of visual short-term memory in monkeys and the spatial properties of images. AB - Experiments were performed on Rhesus macaques to study the relationship between delayed visual differentiation processes and stimulus properties. These investigations showed that the processes of short-term storage of visual information in monkeys has significant features associated with differences in stimulus properties. These consisted of different durations of storage and motor response times. Because of these differences, stimuli (15 pairs) could be grouped into compact clusters on the bases of similarity between their delayed differentiation characteristics. These experiments characterized the processes of short-term information storage during the differentiation of stimuli differing in terms of spatial relationships between elements, as compared with stimuli differing in terms of other attributes (shape, color, etc.); spatial information was stored for shorter periods of time and motor response times were longer. It is suggested that visual short-term memory involves a set of mechanisms operating on attributes of different types and which, along with signs and working programs associated with the visual system, stores spatial discriminatory signs, in which the major role is played by visual-vestibular interactions. PMID- 9850967 TI - Restoration of locomotion in white rats after multiple lesioning of the motor cortex and heterotopic transplantation of cortex fragments. PMID- 9850970 TI - [The acetabular labrum and labrum injury]. PMID- 9850969 TI - Projections of the amygdaloid body, ventral tegmental area, and substantia nigra to various segments of the nucleus accumbens in the dog brain. PMID- 9850968 TI - Morphofunctional specialization of the main and accessory magnocellular neuroendocrine nuclei of the hypothalamus. PMID- 9850971 TI - [The acetabular labrum in infants]. AB - The knowledge of embryology and early development of the hip joint (and especially the "Anlage" of the acetabular labrum) is necessary to correctly understand further growth disturbances and developmental dysplasias and dislocations of the hip joint. "Teratologic" luxations--based on damages of the fetal "Anlage"--should be distinguished from "developing deformations" of originally normal-shaped hip joints. By using modern imaging techniques, especially sonography and MRI, the morphologic relationships of the acetabular labrum in centered and decentered hip joints could be clarified. The tip and the basis of the labrum and their relations with the hyalin-preformed growth zone cartilage of the acetabular roof are changing during the process of decentering and can be clearly visualized by sonographic means. To understand the morphologic changes in decentered hip joints, a clear and consistent terminology should be used: The term "limbus" is misleading and should be avoided; one should use the terms "acetabular labrum" and "hyalin-preformed cartilaginous acetabular roof" for the two histologic subdivisions of the acetabular roof cartilage. We do not know much about size and shape, about nutrition and vascularity of the acetabular labrum, and only few facts about the junction zone of the labral attachment to the hyalin cartilage acetabular roof, and so on. We also do not have any clear evidence about possible micro-damages of the labral-capsular-complex during successful closed reduction and their sequelae in childhood and adolescence, possibly being one cause of labral lesions in adults. Further basic research in this field seems useful and necessary. PMID- 9850972 TI - [Anatomy of the labro-capsular complex]. AB - The acetabular labrum appears as a bundle of distinctly circular lined up collagenous fibers. It surrounds the limbus tangentially and is separated from the cartilagenous covered facies lunata through a thin gap (fissure) except of a small zone in the craniocaudal part. The labrum is strongly fixed with the transverse acetabular ligament. A vascular anastomotic ring surrounds the capsular attachment. It derives its blood supply especially from the superior gluteal vessels, the obturator artery and one ascending branch of the medial femoral circumflex artery. The innervation of the acetabular labrum is coming from a branch of the nerve to the quadratus femoris muscle and from the obturator nerve. There are all types of mechanoreceptors in the labrum. The acetabular labrum is able to exert a high tensional force on the rim of the acetabulum. This plays a very important role in view of the physiological, load depending incongruity of the articulating parts of the hip-joint. PMID- 9850973 TI - [Clinical and diagnostic imaging of labrum lesions in the hip joint]. AB - Labral lesions are common findings in residual hip dysplasia, indicating biomechanical decompensation of the hip joint. MR-Arthrography has shown an excellent accuracy of over 90% to detect these lesions. Nevertheless, so far clinical sings, radiological diagnosis and therapeutical consequences are not well known. In a prospective study, patients suspicious for labral lesions were evaluated using a standard clinical protocol, including history, clinical signs, radiography and MR-Arthrography. Clinical signs were tested by six criteria and two provocation tests. In 11% patients clinical suspicious was wrong. Best agreement with MR-Arthrography was found for "knife sharp" groin pain (100%), impingement test (100%) and painful giving way (83%). 35% of patients showed minor (grade 2) and 52% severe (grade 3 and 4) dysplasia. Independent from grade of dysplasia, no or only slight arthrosis (grade 0 and 1) was found in 64% of patients. In 16% a single acetabular cyst could be detected on radiographs, which all could be identified as intraosseous ganglia on MR-Arthrography. Labral lesions type A (post traumatic) were found in 23% of the patients with only no or minor dysplasis (grade 1 and 2), whereas labral lesions type B (dysplastic) were found in 67% of the patients with severe dysplasia (grade 3 and 4). Clinical signs for labral lesions are typical but can also be observed in other pathologies of the hip joint. Based on the findings of this study, we recommend radiographic evaluation for dysplasia and MR-Arthrography in patients with clinical suspicion for labral lesions of the hip joint. PMID- 9850974 TI - [MRI arthrography in labrum lesions of the hip joint. Method and diagnostic value]. AB - Magnetic Resonance imaging with its excellent soft tissue contrast represents the method of choice for examination of intraarticular and periarticular structures of joints. However, conventional MR-imaging without intraarticular contrast application is not suitable for the diagnosis of labral lesions. MR-arthrography (MRA) of the hip clearly demonstrates labral pathology by distension of the joint space and entrance of the contrast medium into the lesion. Multiplanar 3-D gradient echo sequences enable visualisation of the most important cranio-ventral portion of the labrum. With the MRA labral lesions can be subdivided into different, clinically relevant stages. Furthermore accompanying lesions (intra- and extraosseous ganglia, stress-bone marrow-edema) can be demonstrated. In our study including operatively verified cases, MRA showed excellent sensitivity (90%) and accuracy (91%) in detecting labral lesions. So far no complication occurred with more than 100 MRA's. Therefore this technique represents the method of choice for the radiological evaluation of labral lesions of the hip joint. In this review the technical details and results of our MRA method will be presented. PMID- 9850975 TI - [Labrum lesions from the viewpoint of arthroscopic hip surgery]. AB - The torn acetabular labrum is a classical indication for hip arthroscopy. Clinical and imaging diagnosis are not so reliable compared to hip arthroscopy, which also allows therapeutic intervention. Advances in the technique of arthroscopic examination of the hip joint enable an accurate diagnosis of a torn acetabular labrum as well as arthroscopic resection of such pathology. During 267 consecutive hip arthroscopies, 37 labral lesions (13.9%) could be identified and were resected successfully. The morphological classification of labral tears, and their frequency was allocated to radial flap tears (57%), fibrillated radial tears (22%), longitudinal peripheral tears (16%) and unstable labra (5%). One year after surgery, 78% of the patients were improved, whereas 5 were not improved and 3 had worsened. Larger series with longer follow-up should be reported enabling scientific assessment of the procedure. At this early stage, however, we would recommend that all patients with a suspected labral tear should undergo hip arthroscopy. PMID- 9850976 TI - [Legal grounds for patient information in orthopedics]. PMID- 9850977 TI - [Ligament injuries of the foot. Diagnosis and therapy]. PMID- 9850978 TI - CSF N-CAM in neuroleptic-naive first-episode patients with schizophrenia. AB - An increased concentration of neural cell adhesion molecule (N-CAM) 105-115 kDa has been reported in patients with schizophrenia in both CSF and in post-mortem brain samples. To determine whether increased N-CAM is integral to the disease process or, alternatively, results from early treatment, CSF N-CAM was measured in a blind study of first episode (FE) patients, who were either neuroleptic naive (NN) or neuroleptic-treated (NT, < 100 mg Haldol equivalents), multi episode (ME) patients, and controls. Overall, the FE patients displayed lower N CAM concentrations as compared to controls (p = 0.043). This decrease in N-CAM in FE patients was seen only in the FE-NT group as compared to both controls (p = 0.0006). The FE-NT group also showed a lower CSF N-CAM compared to that in the FE NN (p = 0.025) group. No difference in CSF N-CAM between the FE-NN and control group was found. ME patients showed an increased N-CAM as compared with FE patients (p = 0.018), but not as compared to controls (p = 0.93). Neuroleptic naive first-episode patients do not display a phenotypic increase in N-CAM. Thus, N-CAM is altered in first-episode patients following acute neuroleptic treatment and withdrawal, as compared to neuroleptic-naive first-episode patients. PMID- 9850979 TI - Differential amygdala activation in schizophrenia during sadness. AB - Several studies have reported impaired emotion processing in schizophrenic patients. However, the corresponding functional cerebral correlates of such impairment have not been fully understood, leaving the neurobiological basis of their affective symptoms unknown. Functional magnetic resonance imaging (fMRI) was utilized to examine brain activity in subcortical and cortical regions of 13 medicated male schizophrenic patients and 13 matched healthy controls during happy and sad mood induction. Results show brain activity in the amygdala of normal controls during negative affect, which is in line with previous neuroimaging findings. Unlike controls, schizophrenic patients have not demonstrated amygdala activation during sadness despite matched ratings to normal controls indicating a similar negative affect. Recognizing that structural abnormalities exist in the amygdala of schizophrenic patients, our results provide new evidence of functional abnormalities in the limbic system. PMID- 9850980 TI - Seasonal influences on admissions in schizophrenia and affective disorder in Ireland. AB - Although the seasonal patterns of admissions of affective disorder have been extensively studied, less attention has been given to the seasonal admission patterns of schizophrenia. The traditional method of aggregating the data over a study period, rather than analysing by year of admission may obscure potentially relevant fluctuations in the seasonal pattern. We examined the year-to-year variation in the admission patterns of schizophrenia and affective disorder in Ireland. Using the National Psychiatric Inpatient Reporting System (NPIRS), individuals admitted with an ICD-9/10 diagnosis of a first episode of schizophrenia or affective disorder during the 6-year period 1989-1994 were identified. Seasonal variations in their admission patterns were examined statistically and graphically. There was a significant seasonal variation in the monthly admission patterns of both schizophrenia and affective disorder. This pattern was more marked for individuals with affective disorder. However, the seasonal pattern was not constant from year to year, particularly for schizophrenia. PMID- 9850981 TI - The composition of the depressive syndrome in acute schizophrenia. AB - The composition of the depressive syndrome was examined at both the acute and chronic phases of schizophrenic illness in 86 newly admitted patients. A subgroup with pronounced depression was defined, and a discriminant analysis was performed using symptoms from the Hamilton Rating Scale for Depression (HRSD) as discriminant variables. At the acute phase, the following nine symptoms from the HRSD were significant: depressed mood, guilt, suicide, retardation, three types of insomnia, and two somatic symptoms. At the chronic stable phase, only four symptoms were significant: depressed mood, suicide, general somatic symptoms, and loss of weight. Initial insomnia, middle insomnia, genital symptoms, and loss of insight were poorly correlated. The positive and negative symptoms and extrapyramidal side-effects were not discriminators at either phase. These findings suggest that only certain items from the HRSD may be crucial when assessing depression in schizophrenia. PMID- 9850982 TI - Expressed emotion of families and negative/depressive symptoms in schizophrenia: a cohort study in Japan. AB - This study investigated whether the Expressed Emotion (EE) status of families is associated with an increased risk of negative and depressive symptoms in schizophrenia. The subjects were 52 schizophrenic patients from mental hospitals in Kochi, Japan, who satisfied the DSM-III-R or ICD-9 criteria for schizophrenia. The Japanese version of the Camberwell Family Interview was administered to 73 key relatives of the patients within 2 weeks after admission. A certified rater evaluated the EE's status of each family, using an audiotaped interview and its transcript. Using cohort study design, the subjects were followed for 9 months after their discharge and were monitored for negative and depressive symptoms. Trained psychiatrists who were blind to the EE status of the patients' households administered the Brief Psychiatric Rating scale (BPRS) at discharge, and 9 months after discharge. Repeated-measures analyses of variance showed that EE effects were significant in Emotional withdrawal in all subjects, Depressive mood and Total depressive symptoms in non-relapsers. A multiple regression analyses revealed that EE's effect was significant only in Depressive mood in non relapsers controlling confounding factors. In non-relapsers in positive symptoms, high EE families could make patients depressed. EE's effect on negative symptoms remains to be established. PMID- 9850983 TI - Burden and well-being of caregivers for the severely mentally ill: the role of coping style and social support. AB - Caregivers of persons with severe mental illness often experience a significant burden in coping with patients' symptoms. Several factors have been hypothesized to mediate the impact of caring for a mentally ill relative, including cognitive appraisal, coping strategies, and social support. The present study examined the relationships between these factors, and subjective burden and well-being in caregivers of persons with a severe mental illness. Higher levels of subjective burden were related to (1) greater perceived frequency of positive and negative symptom behaviors, (2) a tendency to use problem-focused oriented coping for dealing with negative symptom behaviors, and (3) a tendency not to use problem solving oriented coping for dealing with positive symptom behaviors. Well-being was also related to lower perceived frequency of positive symptom behaviors and social support, but not to coping style. The implications of the findings for interventions designed to reduce caregiver subjective burden are discussed. PMID- 9850984 TI - Recruitment of non-patient volunteers with schizophrenia spectrum personality symptoms. AB - Authors aimed to evaluate the yield and effectiveness of recruiting community schizophrenia spectrum personality (SSP) subjects via targeted newspaper advertisements listing SSP traits. Eight newspaper advertisements listing SSP traits were placed in regional newspapers over a 3-year period. Respondents (n = 209) were screened thoroughly via telephone, and eligible subjects were invited for face-to-face clinical interviews. One hundred and one subjects (48% of the respondents) were screened out over the phone and another 30% were no-shows or refused. Of the 46 subjects who were interviewed, a majority (24 subjects; 52%) met this study's criteria for SSP. These subjects experienced significant psychotic-like symptoms, as ascertained by a self-rating scale, and showed a downward drift in their socio-economic status. One can successfully recruit SSP subjects, with high yield, using targeted telephone advertisements combined with thorough telephone screening. PMID- 9850986 TI - Covert visual attention in patients with early-onset schizophrenia. AB - The aim of the present study is to examine attentional costs (inhibition) in covert visual attention in a group of acutely ill adolescents with schizophrenia without long histories of neuroleptic treatment. Variations in reaction time were analyzed for possible age and sex differences. Adolescents with schizophrenia (n = 19) were compared to a group of ADHD subjects (n = 20) and a group of normally functioning adolescents (n = 30) on a measure of covert visual attention. The results support a hypothesis of abnormally rapid disengagement (reduced costs) in male adolescents with schizophrenia. Such an abnormality has also been found in adults with chronic schizophrenia. Whether this holds true for both sexes of adolescents with schizophrenia or is restricted to male subjects cannot be answered with certainty due to the small number of females with schizophrenia in our sample. Our findings indicate, however, that there are some general sex differences and some specific sex differences related to covert visual attention in adolescents with schizophrenia. PMID- 9850985 TI - Deficient cancellation of the vestibular ocular reflex in schizophrenia. AB - Schizophrenia has long been associated with difficulties in visual tracking of a moving object. Deficits are most notable in tracking tasks that require inhibition of saccades during active smooth pursuit. In order to assess whether there is a more global problem in inhibition of other eye movement systems while the smooth pursuit system is active, this study examined cancellation of the vestibular ocular reflex (VOR). Cancellation of the VOR occurs in a task in which the subject is rotated while looking at a target that is also being rotated. This requires the subject to use the pursuit system to override the VOR, maintain the eye at a stable location within the orbit, and thus retain visual gaze upon the target. Thirteen individuals with schizophrenia and 15 normals were assessed during clockwise rotation at 60 degrees s-1. Schizophrenic subjects had a significant increase in counterclockwise slow velocity eye movements, suggesting an impaired ability to cancel the VOR. Cancellation of the VOR is thus another example of a breakthrough of an alternative eye movement system while the smooth pursuit system is active. Because of the simplicity of the VOR and its suitability for animal modeling, investigation of this phenomenon may delineate more precisely the mechanisms of visual tracking dysfunction in schizophrenia. PMID- 9850987 TI - Dopamine receptor D2 Ser/Cys311 variant associated with disorganized symptomatology of schizophrenia. AB - The dopamine D2 receptor gene has been proposed as a genetic risk factor for schizophrenia (Arinami et al., 1994). However, a number of replications failed to confirm the initial report. The finding of a stronger association considering schizophrenics with the absence of negative symptoms (Arinami et al., 1996) suggested that the influence of DRD2 variants should be analyzed more at the level of symptoms rather than syndromes. One hundred and four inpatients affected by schizophrenia (n = 99) and delusional disorder (n = 5) (DSM IV) were assessed at admission by the Operational Criteria for Psychotic Illness (OPCRIT) and were typed for DRD2 variants using polymerase chain reaction (PCR) techniques. Subjects with the S311C variant presented a higher score on the 'Disorganization' factor (P = 0.012). Consideration of possible stratification effects such as sex and age of onset did not reveal any deviation from the whole sample. In conclusion, our preliminary report suggests that the DRD2 S311C variant may be a liability factor for disorganized symptoms among schizophrenics or for a subtype of schizophrenia characterized by highly disorganized symptomatology. PMID- 9850988 TI - Increase in urinary peptides prior to the diagnosis of schizophrenia. PMID- 9850989 TI - Detection of antibodies to bovine viral diarrhoea virus (BVDV) and characterization of genomes of BVDV from Brazil. AB - An ELISA for the detection of antibodies to bovine viral diarrhea virus (BVDV) was developed based on antigens derived from a genotype I BVDV strain isolated in Switzerland. Using monoclonal antibodies we showed that this antigen contained the conserved non-structural protein NS3 whereas it essentially lacked the more strain-specific E2 surface glycoprotein. This ELISA has a sensitivity of 97.5% and a specificity of 99.2% as compared to the serum neutralization test (SNT). Preliminary experiments showed that this ELISA reliably detects antibodies to BVDV strains circulating in Brazil. Serum samples obtained from 430 adult cattle on 19 farms of the State of Rio Grande do Sul (Brazil) and one farm from Corrientes (Argentina) were tested for antibodies by means of this ELISA. We found antibodies in 56% +/- 15.1% of the cattle sera tested, which indicates that, in Brazil, the prevalence of infection with BVDV is similar to that found in Europe and the USA. Our sequence analysis of two BVDV isolates showed that BVDV of both genotypes I and II circulate in Brazil. PMID- 9850990 TI - Pseudorabies virus (PRV) early protein 0 activates PRV gene transcription in combination with the immediate-early protein IE180 and enhances the infectivity of PRV genomic DNA. AB - Pseudorabies virus (PRV) early protein 0 (EP0) functions as a transactivator of the viral gene promoters. In transient expression assays employing chloramphenicol acetyl transferase (CAT) reporter constructs, EP0 and the immediate-early protein IE180 act in an additive manner to activate transcription from the thymidine kinase (TK) and glycoprotein G (gG) gene promoters. EP0 enhanced the synthesis of infectious virus in cotransfection experiments with the EP0-expression plasmid and PRV genomic DNA. EP0 was detected by Western blot analysis in the purified virions. These results may indicate that EP0 in the virions acts as an important transactivator to express the immediate-early gene efficiently in the first stage of infection, and IE180 and EP0 expressed after the infection cooperatively activate the early and late gene expression in the later stage of infection. PMID- 9850991 TI - Asymptomatic bacteriuria in puppies with canine parvovirus infection: a cohort study. AB - This study aimed to investigate the possible association between canine parvoviral enteritis and asymptomatic bacteriuria. Forty-three puppies that were admitted to the outpatient service of the Animal Medical Clinic with clinical signs compatible with parvoviral enteritis formed the exposed group. The clinical diagnosis was subsequently confirmed by a positive fecal ELISA test (CITE test: IDDEX Lab., Westbrook, ME). Twenty-three (53.5%) of these puppies were males and 20 (46.5%) were females. Their age ranged from 1.5 to 5.5 months. Forty-eight clinically normal and age-matched puppies, that had been admitted to the clinic for vaccinations and had a negative result in the aforementioned ELISA test, were randomly selected to form the unexposed group. Urine samples were collected by antebupic cystocentesis from all puppies and submitted for bacterial culture. In the parvovirus exposed group, 11 of 43 puppies had detectable bacteriuria. The isolates were Escherichia coli alone (8/11-72.7%) Staphylococcus aureus alone (1/11-9.1%) and mixed cultures of E. coli and S. epidermitis (2/11-18.2%). In the unexposed group there were three puppies with detectable bacteriuria, one isolate each of E. coli, Enterococcus durans and Corynebacterium spp. Puppies with parvoviral enteritis had five (95% CI: 1.3-19.8) times higher odds of developing asymptomatic bacteriuria than puppies without the disease. The observed increased risk of asymptomatic bacteriuria among puppies with parvoviral enteritis was probably due to the fecal contamination of the external genitalia and the neutropenia these puppies exhibited. PMID- 9850992 TI - A stable cell line with a proportion of cells persistently infected with bovine viral diarrhoea virus. AB - Bovine turbinate (BTu) and lamb testis (LT) cell lines persistently infected with bovine viral diarrhoea virus (BVDV) arose as a result of a single change of medium containing commercial foetal calf serum. Infected cells comprise 30% and 50% respectively, of the total cell population, determined by immunohistochemical staining. The ratio of positive cells has remained unchanged during successive passages. Characterization of the persistently-infected BTu cells (BTuI) showed that only full length viral RNA was detected by northern blot hybridisation, indicating that DI particles were not involved. Secreted and intracellular virus from these cells was fully infectious for fresh BTu and LT cells. The BTuI cell line was fully permissive for a cytopathic BVDV isolate and a bovine herpesvirus, but non-permissive for two non-cytopathic BVDV isolates. Attempts to induce the permissive state in the BVDV-negative cells of the BTuI culture by treatment with actinomycin D and 5'-aza-cytidine failed. These cells provide a convenient model to study aspects of BVDV pathogenesis and replication. PMID- 9850993 TI - Monoclonal antibodies against conformationally dependent epitopes on porcine reproductive and respiratory syndrome virus. AB - Monoclonal antibodies (MAbs) against porcine reproductive and respiratory syndrome virus (PRRSV) were prepared and characterized. Four MAbs were developed from the mice immunized with the recombinant GP4 protein expressed in insect cells, and six MAbs were derived from the immunization with recombinant GP5 protein. All of the MAbs showed strong perinuclear fluorescence in PRRSV VR2385 infected cells by immunofluorescence staining. Among the MAbs to GP5 protein, one showed strong reactivity in ELISA and recognized a 26 kDa band of PRRSV in a western blot assay, while another showed neutralizing activity against the VR2385 isolate. Out of the four MAbs to GP4 protein, one showed mild reactivity in ELISA with detergent extracted antigen, but had no reactivity in a western-blot assay. The failure of MAb binding to detergent extracted antigen in ELISA or in western blot analysis indicated that the MAbs were against conformationally dependent epitopes. Reactivity patterns of the MAbs with PRRSV field isolates tested by fixed-cell ELISA showed that there are antigenic variations in PRRSV GP4 and GP5 proteins. Development of these MAbs will benefit further studies on PRRSV structural proteins as well as in understanding their roles in PRRSV pathogenesis. PMID- 9850994 TI - In vitro studies on the use of clay, clay minerals and charcoal to adsorb bovine rotavirus and bovine coronavirus. AB - Rotaviruses are the leading cause and coronaviruses are the major contributors of acute gastroenteritis in the young of various mammalian and avian species. Despite numerous trials and decades of research, vaccines have limited efficacy particularly for calves. As an alternative method of controlling infection, we have investigated broad spectrum antiviral agents that are not discriminatory among various viruses. This report involves testing a variety of adsorbent agents including charcoal, clay, and clay minerals to adsorb rotavirus and coronavirus in vitro. Results revealed that all the adsorbent agents had good to excellent capability of adsorbing rotavirus and excellent capability of adsorbing coronavirus. Percent adsorptions ranged from 78.74% to 99.89% for rotavirus and 99.99% for coronavirus; while sand (negative control) was < 0.01%. A high affinity binding was present as determined by a low percent desorption (0.06 3.09%). However, the adsorbent bound virus complex retained, and may have actually enhanced, infectivity. PMID- 9850995 TI - An in vitro study of theaflavins extracted from black tea to neutralize bovine rotavirus and bovine coronavirus infections. AB - Crude theaflavin was extracted from black tea and then fractionated by HPLC into five components (initial peaks (IP), TF1, TF2A, TF2B, and TF3). The crude extract and the various fractions of theaflavin were collected and tested, individually and in combination, for antirotaviral activity. The mean effective concentration (EC50) was calculated and compared. Activity varied from the most active being the uncharacterized theaflavin-like initial peaks (IP) with an EC50 of 0.125 microgram/ml to the least active being theaflavin-3 monogallate (TF2A) with an EC50 of 251.39 micrograms/ ml. The combination of TF1 + TF2A + TF2B + TF3 was more active than the sum of the activities of these four fractions individually, indicating synergism among the peaks. Only the crude extract was assayed for activity against coronavirus; the EC50 was 34.7 micrograms/ml. PMID- 9850997 TI - Influence of pig age on virus titer and bactericidal activity of porcine reproductive and respiratory syndrome virus (PRRSV)-infected pulmonary intravascular macrophages (PIMs). AB - Twelve pigs (six 4-week-old and six 4-month-old cross-bred, specific pathogen free pigs) were used as donors for both pulmonary intravascular macrophages (PIMs) and pulmonary alveolar macrophages (PAMs). The PIMs and PAMs were infected in vitro with low (ISU-55) or high (VR-2385) virulence strains of PRRSV at 1 multiplicity of infection (m.o.i.) for comparisons of virus titers at 48 h post infection (PI). PIMs were as permissive as PAMs to infection with both PRRSV isolates yielding similar progeny titers (10(4.81) vs. 10(5.22) TCID50/ml, respectively). Both ISU-55 and VR-2385 were able to infect PIMs and no significant difference in virus replication as measured by virus titers between isolates was found (10(5.33) vs. 10(4.69) TCID50/ml, respectively). PIMs from 4 weak-old pigs yielded a higher virus titer following PRRSV infection than PIMs from 4-month-old pigs (10(5.43) vs. 10(4.59) TCID50/ml, respectively; p < 0.02). VR-2385-infected PIMs had significantly decreased bactericidal (Staphylococcus aureus) activity compared with uninfected PIMS at 48 h PI (p < 0.05). There was no difference in bactericidal activity between ISU-55 (low virulence)-infected PIMs and VR-2385 (high virulence)-infected PIMs. Both ISU-55 and VR-2385 infection significantly decreased the production of superoxide anion (SOA) at 24 and 48 h PI (p < 0.01). In conclusion, (1) PRRSV had a detrimental effect on bactericidal activity and SOA production of PIMs, (2) PIMs from younger pigs were more permissive to PRRSV infection, and (3) the selected PRRSV strains, which differ in their abilities to induce pneumonia in vivo were not different when tested in vitro by measuring virus titer and bactericidal functions. PMID- 9850996 TI - A bovine respiratory syncytial virus strain with mutations in subgroup-specific antigenic domains of the G protein induces partial heterologous protection in cattle. AB - Bovine respiratory syncytial virus (BRSV) strains are tentatively divided in subgroups A, AB and B, based on antigenic differences of the G protein. A Dutch BRSV strain (Waiboerhoeve: WBH), could not be assigned to one of the subgroups, because the strain did not react with any monoclonal antibody against the G protein. We describe here that the WBH strain has accumulated critical mutations in subgroup-specific domains of the G protein gene, which also occur but then independently in G protein genes of BRSV subgroup A or B strains. Although the comparison of nucleotide residues 256-792 of the G gene of the WBH strain with those of subgroup A and B strains showed that the G gene of the WBH strain is different from that of BRSV subgroup A and B strains, the sequence divergence was not more than observed within the G genes of human respiratory syncytial virus subgroup A or B strains. The WBH strain did not induce severe disease after experimental infection of calves, and induced partial protection against a heterologous challenge. Despite the dissimilarity of the conserved central regions of the G protein of the WBH strain and that of the challenge strain, a secondary antibody response against this region was induced in WBH-infected calves after challenge. We conclude that complete BRSV virus can partially protect against a BRSV infection with a strain that contains an antigenic dissimilar G protein. PMID- 9850998 TI - Intraspecies polymorphism of vsp genes and expression profiles of variable surface protein antigens (Vsps) in field isolates of Mycoplasma bovis. AB - To assess the extent of interstrain variation, 50 isolates of Mycoplasma (M.) bovis including the type strain PG45 were examined for the presence of a family of variable membrane surface lipoproteins (Vsps) and their genes. Southern hybridization using a genomic fragment carrying three distinct vsp genes (vspAEF) revealed a striking heterogeneity, with only 2/50 strains having identical banding patterns. Cluster analysis of the data showed that most isolates from interrelated herds (groups 1, 2 and 3) were combined in a cluster of 50% homology, while isolates from distinct geographical regions (groups 4, 5 and 6) were linked only at 18% homology. Vsp antigen expression was monitored by Western immunoblotting using four specific monoclonal antibodies (MAbs). Resembling the findings at the DNA level, interstrain variation of Vsp expression among groups 1 3 was less pronounced than among non-interrelated isolates from groups 4-6. Ten out of 50 strains did not hybridize with the vspAEF gene probe at high-stringency conditions, 8/50 failed to react with any of the Vsp-related MAbs, and 6/50 proved negative in both assays. Interestingly, most of these isolates produced hybridization signals at low stringency suggesting major distinctions in their vsp gene structure. The extensive evidence obtained on interstrain vsp gene polymorphism and variation in Vsp expression could provide a basis for a future understanding of the pathogenic potential of individual M. bovis strains. PMID- 9850999 TI - Acute septicaemic pasteurellosis in Vietnamese pigs. AB - Sixteen isolates of Pasteurella multocida were cultured from cases diagnosed as acute septicaemic pasteurellosis in Vietnamese pigs. The HSB-PCR assay provided rapid presumptive determination of 10 isolates of P. multocida identified as haemorrhagic septicaemia (HS) causing type B cultures (B:2, B:5, B:2,5). Serological designation using the Carter and Heddleston typing systems confirmed these findings, and identified the six HSB-PCR negative cultures as either A:1, A:3 or D:3,4. Biochemical fermentation and REP-PCR revealed phenotypic and genotypic identity between P. multocida type A:1 isolated from Vietnamese pigs and poultry. Marked homogeneity was also demonstrated among HSB-PCR positive swine isolates, which were shown to possess genotypic identity with P. multocida type B:2 from buffaloes diagnosed with HS. PMID- 9851000 TI - Tetracycline resistance in Staphylococcus spp. from domestic animals. AB - A total of 838 staphylococcal isolates representing 19 different species were obtained from cattle, cats, dogs, ducks, guinea pigs, horses, mink, pigeons, pigs, rabbits, and turkeys. From these 228 (27.2%) isolates were shown to be resistant to tetracycline and to carry one or two of the tetracycline resistance (tet) genes tet (K), tet (L), tet (M), or tet (O) with seven different distribution patterns. Additional resistances to one or more antibiotics were observed in 153 (67.1%) of the tetracycline resistant isolates. The tet (M) gene was found in 94.3% of the resistant S. intermedius isolates while the tet (K) gene predominated in most of the other staphylococcal species irrespective of the host animal. The tet (K) and tet (L) genes were located on plasmids while the tet (M) and tet (O) genes appeared to be associated with the chromosome. PMID- 9851001 TI - Simultaneous flow cytometric measurement of Streptococcus suis phagocytosis by polymorphonuclear and mononuclear blood leukocytes. AB - A simple flow cytometric method was used to study simultaneously the phagocytosis of Streptococcus suis serotype 2 by polymorphonuclear and mononuclear blood leukocytes from swine and humans. Using this method with a bacteria-to-leukocytes ratio of 10:1 and after 60 min of incubation, 80.2 +/- 2.8% of swine granulocytes and 77.0 +/- 2.8% of swine monocytes were shown to contain FITC-labelled S. suis serotype 2 strain 735. Using the same strain, FITC-labelled bacteria were found in 95.5 +/- 3.2% of human granulocytes and in 92.8 +/- 3.6% of human monocytes. The phagocytosis rates of avirulent and virulent strains of S. suis were not significantly different. PMID- 9851002 TI - Verotoxin-producing Escherichia coli (VTEC) and eae-positive non-VTEC in 1-30 days-old diarrhoeic dairy calves. AB - Faecal samples from 221, 1-30-days-old, diarrhoeic dairy calves were screened for the presence of verotoxin-producing Escherichia coli (VTEC) and eae-positive non VTEC. Calves were grouped according to their age (1-7, 8-14, 15-21 and 22-30 days) and analyses of prevalences were done by Mantel-Haenzsel chi 2-test for trend. VTEC and eae-positive non-VTEC were detected in 20 (9.0%) and 18 (8.1%) of the diarrhoeic calves, respectively. A significant age-associated increase in the prevalence of VTEC (p = 0.0001), but not in the prevalence of eae-positive non VTEC (p = 0.381), was found. Significant differences in VTEC prevalence were found between the age-group 22-30 days and in all other age-groups. 43 (5.0%) of the 861 E. coli isolates from the 221 diarrhoeic calves were VTEC, and 30 (69.8%) of these strains produced VT1 only. More than one-half of the VTEC strains (55.8%) were positive for the eae gene and all these eae-positive VTEC strains produced VT1 only. A high percentage (76.7%) of VTEC strains belonged to E. coli serogroups (O4, O26, O39, O91, O113, O128 and O145) associated with haemorrhagic colitis and haemolytic uraemic syndrome in humans. 51 (5.9%) of the E. coli strains studied were eae-positive non-VTEC and the serogroups most prevalent among these strains were O4, O14, O26 and O123. Only four of the eae-positive strains were also espB-positive by hybridization with a probe from a human EPEC isolate and none of these strains produced VT. PMID- 9851004 TI - Characterization of mechanisms involved in uptake of Streptococcus dysgalactiae by bovine mammary epithelial cells. AB - Bovine mammary epithelial cells were pretreated with inhibitors of protein kinase activity, actin polymerization and receptor-mediated endocytosis. In addition, mammary epithelial cells and Streptococcus dysgalactiae were pretreated with inhibitors of protein synthesis. Results showed that activity of tyrosine protein kinases, intact microfilaments and de novo eukaryotic protein synthesis was required for uptake of S. dysgalactiae by bovine mammary epithelial cells; a process that appeared to occur via receptor-mediated endocytosis. In contrast, de novo bacterial protein synthesis was not required for uptake of S. dysgalactiae by MAC-T cells. This study provides insight into bacterial and cellular mechanisms involved in early host-pathogen interactions, putting into perspective the role of mammary epithelial cells in the development and establishment of intramammary infections by S. dysgalactiae. PMID- 9851003 TI - Prevalence and molecular typing of attaching and effacing Escherichia coli among calf populations in Belgium. AB - Attaching and effacing Escherichia coli are involved in diarrhea in 2 to 8-week old calves. The virulence factors of these bacteria include: (i) the secretion of proteins (i.e. EspB) involved in microvilli effacement, (ii) the production of the intimin, a 94 kDa outer membrane protein encoded by the eaeA gene and involved in the intimate attachment of bacteria to epithelial cell and (iii) the production of verotoxins: VT1 and/or VT2. We investigated the presence and the pathotype of these strains in several calf populations by colony hybridization or by genetic amplification. Using the colony hybridization method we showed first that only 5% of calves who died from diarrhea presented EaeA+ E. coli strains and secondly that 19% of healthy calves showed an asymptomatic carriage. However, using colony hybridization and genetic amplification, we identified EaeA+ strains in 91% of calves living in farms with recurrent diarrhea problems. In 66% of the calves, there was a correlation between the presence of AEEC and diarrhea. At the pathotype level, most of the EaeA+ isolates were negative for VT probes. In VT+ bacteria, the majority were VT1+. The number of VT positive bacteria was significantly higher in calves who died from diarrhea than in healthy or sick calves. This underlined the aggravating role of verotoxins in the disease. Moreover, only 25% of the bovine AEEC were positive with the EaeB probe. Surprisingly, the proportion of EaeB+ strains was significantly higher in healthy calves than in other populations. PMID- 9851005 TI - The use of pulsed field gel electrophoresis to investigate the epidemiology of Staphylococcus aureus infection in commercial broiler flocks. AB - Pulsed field gel electrophoresis (PFGE) analysis of SmaI restriction patterns was used to type 109 isolates of Staphylococcus aureus collected from broiler farms and hatcheries in Northern Ireland. Forty-seven isolates from clinical conditions in broilers and 62 strains from hatcheries, were examined. The PFGE patterns demonstrated a similarity between 85% of strains from clinical sources and 71% of the hatchery isolates. The association of disease with the predominant strain type and presence of these same strains in the hatchery, indicates that the hatchery is a potential source of the infection for clinical broiler disease. PMID- 9851006 TI - Calculated milk production losses associated with elevated somatic cell counts in dairy cows: review and critical discussion. AB - Relationships between somatic cell count (SCC) and variation in milk production at the cow level were reviewed to provide average reference values suitable for the assessment of economic losses due to subclinical mastitis. The literature analysis involved 19 papers, defining milk yield and/or its composition either at test-day level or at the whole lactation level as statistical unit. Within each type of approach, study populations and designs differed. Regression models implemented also showed large differences. At test-day level, the average trend was a loss of 0.4 kg of milk in primiparous cows and 0.6 kg in multiparous, by each 2-fold increase of SCC above 50,000 cells/mL. At the lactation level, the average trend was a loss of 80 kg of milk in primiparous and 120 kg in multiparous, by each 2-fold increase of the geometric mean of SCC above 50,000 cells/mL. Protein content of milk showed a small increase of 0.15 g/kg (at the test-day level) while fat content showed a small decrease of 0.20 g/kg (both at the test-day and at the lactation level), by each 2-fold increase of SCC. The value of further studies was underlined, especially to provide more accurate quantification of the composition changes associated with elevated SCC, and to improve the imperfect knowledge about the effects of parity and stage of lactation on the studied relationships. PMID- 9851007 TI - [Vectorial competence of Glossina tachinoides Westwood and Glossina palpalis gambiensis Vanderplank infected by Trypanosoma brucei brucei EATRO 1125]. AB - The vectorial competence (VC) of teneral (less than 32 h) Glossina tachinoides Westwood and G. palpalis gambiensis Vanderplank, fed simultaneously on a guinea pig infected with Trypanosoma brucei brucei EATRO 1125, was assessed. Statistical analysis of the experimental results revealed that female G. tachinoides had a significantly higher midgut infection rate than males. Such a sex-related difference was not observed in G. p. gambiensis. Male G. p. gambiensis had higher midgut infection rates than male G. tachinoides. The metacyclic index did not differ between both subspecies, although G. p. gambiensis showed relatively more metacyclic infections than G. tachinoides. A global VC of 0.0242 and 0.0483 was found for G. tachinoides and G. p. gambiensis, respectively. VC did not differ significantly either between sexes or between the two species. However, G. tachinoides more rapidly infected the feeding host than G. p. gambiensis. In all infected flies, the procyclic index value was superior to the metacylic index value, suggesting that the infection is established by an ascending origin. Both large and slender parts of the salivary glands were constantly infected. Longitudinally dividing trypomastigotes of unequal length have been observed in the alimentary canal of the flies. PMID- 9851008 TI - Kinematics of the equine back: a method to study the thoracolumbar flexion extension movements at the trot. AB - This study was conducted to evaluate a method for quantifying the flexion extension movements of the back of horses trotting on a track in the conditions of the clinical lameness examination. Using a 3-D kinematic analysis system, the successive positions of four markers placed at regular intervals between the withers and the tuber sacrale were recorded. To isolate the flexion-extension movements of the back, the positions of these four markers were recomputed in a trunk-related co-ordinate system of the horse. Then, for each frame, the equation of the third-order polynomial that best fitted the position of the four markers was determined. Using these equations, it was then possible to interpolate the vertical displacement of any point situated between the withers and the tuber sacrale. The accuracy of this method was evaluated using three additional markers placed between the four previous ones. The vertical displacements of these three markers were recorded and compared to the interpolated displacements provided by this method. The results showed that the experimental and interpolated values did not present any statistical difference except in the lumbar region. In this region, the general shape of the vertical displacement curve was conserved but magnified by 17%. This study showed that despite the weakness of the range of the flexion-extension movements of the equine back at trot, these movements can be quantified with reliability. This method was then used on a lame horse. The results obtained demonstrated the usefulness of this method to study the flexion extension movements of horses suffering from various locomotor disorders. PMID- 9851009 TI - Effect of dietary insulin on the response of suckling mice enterocytes to Escherichia coli heat-stable enterotoxin. AB - Effect of insulin on the response of suckling mice to the enterotoxigenic Escherichia coli heat-stable enterotoxin (STa) was studied. Four groups (8-10 in each group) of 2-day-old Swiss Webster suckling mice were used. For this study, 5, 10, 25 and 50 micrograms of insulin was given orally to half the mice in each group for 7 days. The rest of the mice in each group were given normal saline as intra-litter controls. After 7 days, a suckling mouse assay in which 1 microgram of STa was given to all mice in insulin-treated and control groups was performed. Enterocyte suspensions were prepared from mice in all groups. Intestinal tissue samples were taken for electron microscopy. Interaction of STa with its putative receptor on the enterocytes was evaluated using indirect immunofluorescence and flow cytometry. The suckling mouse assay revealed a significant increase in the gut weight to body weight ratio in all mice in the insulin-treated groups compared to control mice (P < 0.05). Flow cytometry and indirect immunofluorescence analyses suggested that insulin had an up-regulatory effect on the STa-receptor level. Similarly, insulin was found to increase intestinal brush border membrane differentiation as indicated by the increase in the inward movement of milk particles through the intestinal mucosa. Insulin seems to modify the structure-function of the brush border membrane including the response of suckling mice to STa. This study may provide further insights into the mechanism of STa/receptor interaction, which is a major cause of diarrhea in newborn animals and human infants. PMID- 9851010 TI - Comparison of the effectiveness of two protocols of antirabies bait distribution for foxes (Vulpes vulpes). AB - In a plateau and hill region of France (the Doubs), two protocols of rabies vaccine bait distribution targeted at foxes were compared: helicopter distribution of vaccine baits alone (control zone) and a combined aerial distribution by helicopter with an additional deposit of vaccine baits at fox den entrances by foot (test zone). In the test zone covering an area of 436 km2, baits were distributed by helicopter at a rate of 13.4 baits/km2. Additionally, an average of 11.4 vaccine baits at the entrances of 871 fox dens were terrestrially distributed by 110 persons (9,964 baits). In this test zone, 90% of the young foxes were marked with tetracycline which permitted estimation of the bait consumption; however, only 38% had significant titre of rabies antibodies and less than one fox cub per 2.4 of those having consumed at least one bait were immunized. In the control zone, these percentages were significantly lower: respectively, 35 and 17% and one fox cub per 4.2. The relative lack of benefit between bait uptake and rate of immunological response may be due to maternal immunity which could have interfered with fox cub active immunization. A booster effect following a second distribution of baits by foot may be suggested in both adult foxes and their offspring. That these baits needed to be terrestrially distributed in order to obtain a booster effect is uncertain. Terrestrial distribution at fox den entrances is difficult to do and entails additional expenses not incurred in aerial distribution. The cost of terrestrial vaccination is 3.5 times higher than classical aerial vaccination and takes 63.5 times longer. A cost effective analysis of this type of supplementary terrestrial intervention determined that bait deposit at den entrances can be recommended for restricted areas, where residual focii exist, as a complement to the aerial distribution of baits. PMID- 9851011 TI - High hydrostatic pressure inactivation of Salmonella typhimurium: effects of pressure, duration, pH and temperature studied by analysis of variance. AB - High hydrostatic pressure treatments are regarded as possible alternative methods for food preservation. One of the primary considerations for industrial applications is the ability of these methods to eradicate pathogenic microorganisms. This study subjected S. typhimurium suspensions, first in a phosphate buffer (pH 7.0) and then in a citrate phosphate buffer (pH 5.6), to high hydrostatic pressure treatments relative to the following variables: pressure (200-400 MPa), duration (3, 10 and 20 min), temperature (4, 20 and 40 degrees C) and the pH of the suspension medium (5.6 and 7.0). An optimal design of 40 runs was obtained using the Fedorov algorithm, and responses were studied by analysis of variance in terms of cell survival on plate count agar. Efficiency was determined by Log10 comparisons of the numbers of live cells before and after treatment. A statistically significant relationship was found between the four variables considered (pressure, pH, duration and temperature), their interactions (duration x pressure, pH x temperature, pH x pressure) and the inactivation of S. typhimurium. R-squared statistical analysis indicated that the linear model used accounted for more than 98% of the variability in the inactivation of S. typhimurium. PMID- 9851012 TI - Comparative relationship between copper-zinc plasma concentrations and superoxide dismutase activity in camels and cows. AB - On an experimental farm, five camels and five cows were fed with a similar basal diet over a 6-month period. They received an oral trace element supplement for 3 months (days 22-112). This supplement contained zinc and copper sulphate, and corresponded to twice the daily requirement generally recommended for cows. Plasma zinc and copper concentrations were significantly lower in camels (44 micrograms/100 mL for copper and 38 micrograms/100 mL for zinc) than in cows (106 and 83 micrograms/100 mL, respectively). The supplementation had no effect on the plasma zinc concentration in the camels in spite of the low observed values. The mean erythrocyte SOD activity was also significantly higher in the cows (2,404 +/ 211 IU/100 gHb) than in the camels (1,720 +/- 312 IU/100 gHb). In both species, no correlation was found between copper plasma concentration and erythrocyte SOD activity. In cows, a positive relationship was observed between plasma zinc concentration and SOD activity (r = 0.396). In contrast, a negative relationship was found in camels (r = -0.369). These results are discussed in relation to the physiological peculiarities of the camel. PMID- 9851013 TI - Absence of porcine interferon alpha secreting cells in severe combined immunodeficiency (SCID) mice inoculated with porcine leukocytes. AB - A low frequency leukocyte subpopulation, referred to as natural interferon producing cells (NIPC) is able to produce high amounts of interferon alpha (IFN alpha) following contact with noninfectious viral structures. In order to examine the possible leukocytic nature and bone marrow origin of NIPC, severe combined immunodeficiency (SCID) mice were reconstituted with porcine leukocyte populations, including bone marrow cells. At different times after reconstitution, enriched CD4 and CD45 positive porcine cells were isolated from various mouse organs and tested for the presence of porcine NIPC by porcine IFN alpha specific ELISPOT assay, after in vitro stimulation by UV inactivated transmissible gastroenteritis virus (TGEV). Although engraftment of porcine cells in SCID mice was shown by flow cytometry and by the production of pig immunoglobulins, no IFN-alpha secreting cells could be detected. This result suggests that NIPC do not derive from bone marrow precursor cells, or that growth factors needed for in vivo expansion of porcine NIPC were absent in mice. PMID- 9851014 TI - Increased severity of acute Trypanosoma brucei brucei infection in rats with alloxan-induced diabetes. AB - Twenty rats were made diabetic by treatment with alloxan monohydrate (10% solution, 100 mg/kg body weight). Ten diabetic and ten non-diabetic rats were intraperitoneally infected with the same infective doses of Trypanosoma brucei brucei (Lafia strain). The uninfected controls were ten diabetic and ten non diabetic rats. The prepatent period was shorter in the diabetics (3.5 +/- 0.5 days) than the non-diabetics (4.2 +/- 0.4 days). Although the infected diabetic and non-diabetic rats had comparable levels of peak parasitaemia, the diabetics had significantly (P < 0.05) higher parasitaemia before this peak. The survival time was shorter (P < 0.05) for the infected diabetics (12.1 +/- 3.2 days) than for the infected non-diabetics (14.8 +/- 1.7 days). The infection did not affect the level of diabetic hyperglycaemia, but caused a more severe anaemia in the diabetics than the non-diabetics, with the percentage decreases in packed cell volume in the diabetics being higher (P < 0.05) from days 3 to 12 post-infection. In conclusion, the pathogenic effects of trypanosome infection may be more severe in rats having alloxan-induced diabetes. PMID- 9851015 TI - [Detection of caprine arthritis encephalitis virus in sperm of experimentally infected bucks]. AB - Shedding of caprine arthritis encephalitis virus (CAEV) was assessed in semen and blood mononuclear cells of six bucks (four boers and two saanens) experimentally contaminated with a viral strain (CAEV Cork) and on three non-infected controls. CAEV was identified by polymerase chain reaction (PCR) in blood mononuclear cells of all infected animals but only in seminal fluid and non-spermatic cells of one buck and in non-spermatic cells of another. Presence of CAEV in semen could have implications in the dissemination and control of the disease. PMID- 9851017 TI - Applications of neural networks to colour recognition. AB - Colour specification and matching are important tasks used in a number of different industries. However, current methods can employ very complicated, elaborate pieces of laboratory equipment which are not portable, such as spectrophotometers with combinations of calibrated light sources and a large number of narrow band filters. Simpler, portable devices are typically not sophisticated enough to give a high degree of quality control due to the less sophisticated light sources and the necessarily smaller number of filters employed. Lastly, techniques such as the 'match by eye' method can lead to deceptive metameric matches. This paper outlines the beginning of the development of a colour measurement system which is portable, easy to use, and more accurate than other portable devices by virtue of an artificial neural network used to analyze the data. With specialized training of the neural network, it could even be used in applications such as teeth and skin colour measurements. PMID- 9851016 TI - Lack of humoral and cellular responses against glutathione S-transferases in Fasciola hepatica experimentally infected sheep. AB - Anti-Fasciola hepatica excretory-secretory products (FhESP) and anti-glutathione S-transferase (GST) immune response were studied in sheep infected with 250 F. hepatica metacercariae. In FhESP, 29 proteins with molecular masses (m) of 10-200 kDa were identified by SDS-PAGE. GST m were 26.7 and 27.2 kDa. The anti-FhESP humoral response was precocious and increased throughout the infection. FhESP contains several immunoreactive antigens but no anti-GST antibodies were detected either by ELISA or western blotting. The anti-FhESP cellular response increased precociously and fell after the second week of infection but no cellular response against GST was observed. PMID- 9851018 TI - Models for the cross-correlation between retinal ganglion cells. AB - Neighboring ganglion cells in the vertebrate retina not only respond to the same stimuli but also display cross-correlated activity on a millisecond time scale. Recent studies of this cross-correlation have indicated that simple linear addition of common variability to each ganglion cell signal does not account for the observations (Levine 1997). In this report, Monte Carlo simulations of various linear and nonlinear models are presented that confirm the earlier speculations. Models in which common variability alters the leakages of a pair of leaky integrate-and-fire neurons account for the data and predict the cross correlogram lag without invoking temporal delay lines. PMID- 9851019 TI - Nonuniformity in the linear network model of the oculomotor integrator produces approximately fractional-order dynamics and more realistic neuron behavior. AB - The oculomotor integrator is a network that is composed of neurons in the medial vestibular nuclei and nuclei prepositus hypoglossi in the brainstem. Those neurons act approximately as fractional integrators of various orders, converting eye velocity commands into signals that are intermediate between velocity and position. The oculomotor integrator has been modeled as a network of linear neural elements, the time constants of which are lengthened by positive feedback through reciprocal inhibition. In this model, in which each neuron reciprocally inhibits its neighbors with the same Gaussian profile, all model neurons behave as identical, first-order, low-pass filters with dynamics that do not match the variable, approximately fractional-order dynamics of the neurons that compose the actual oculomotor integrator. Fractional-order integrators can be approximated by weighted sums of first-order, low-pass filters with diverse, broadly distributed time constants. Dynamic systems analysis reveals that the model integrator indeed has many broadly distributed time constants. However, only one time constant is expressed in the model due to the uniformity of its network connections. If the model network is made nonuniform by removing the reciprocal connections to and from a small number of neurons, then many more time constants are expressed. The dynamics of the neurons in the nonuniform network model are variable, approximately fractional-order, and resemble those of the neurons that compose the actual oculomotor integrator. Completely removing the connections to and from a neuron is equivalent to eliminating it, an operation done previously to demonstrate the robustness of the integrator network model. Ironically, the resulting nonuniform network model, previously supposed to represent a pathological integrator, may in fact represent a healthy integrator containing neurons with realistically variable, approximately fractional-order dynamics. PMID- 9851020 TI - A distributed control model for the air-righting reflex of a cat. AB - A multisegment, multijoint model of a falling animal is presented to examine the effectiveness of a two-stage control scheme in a zero-momentum self-righting maneuver. The model contains a much larger number of degrees of freedom than is required to execute a self-righting maneuver and is thus capable of providing multiple solutions for the same task. The decentralized control scheme is designed to achieve gross turning in minimum time and to maintain a steady orientation relative to gravity after the turn has been achieved. The scheme is able to determine the sequence of steps necessary to execute the motor task and also incorporates learning features. Results from various simulations are presented and their implications discussed. PMID- 9851021 TI - Detection of aperiodic test patterns by pattern specific detectors revealed by subthreshold summation. AB - The hypothesis that the visual system detects, under certain conditions, stimulus patterns by means of filters matched to these patterns (Hauske et al. 1976) may be challenged by the argument that other coding mechanisms like spatial frequency channels, Gabor or Hermite filters mimic the behaviour of matched filters, a view supported by the finding of non-linear contrast-interrelationship functions (CIFs), as determined in superposition experiments. In this paper we argue that an overall non-linear CIF does not contradict the hypothesis of detection by a single matched filter: we find that the sensitivity functions determined in our experiments can be separated into two components reflecting (i) a bandpass filter and (ii) a filter characterized by the spectrum of the test-pattern. PMID- 9851022 TI - Spectral and time domain characteristics of single muscle fibre action potentials during continuous activity extracted from model considerations. AB - A model of the muscle fibre extracellular action potentials (ECAPs) calculation using experimentally recorded intracellular action potentials (ICAPs) has been applied to investigate the effect of repetitive stimulation on the electrical activity of isolated frog muscle fibres. The ECAPs were calculated both at small (0.01 mm) and at large (5 mm) radial distances to the fibre axis, and their relationship with the original ICAP parameters has been inferred. Fourier transformation of the calculated ECAPs in order to obtain the spectral characteristics and to trace out their behaviour during continuous fibre activity was performed. Stimulation frequency dependence on the ECAP time characteristics and on the shift of the maximum spectral density towards low frequencies at small and large radial distance were observed. The spectral density peak frequency is propagation velocity (PV)-dependent. The advantage of the presented method over the available experimental extracellular recording techniques from isolated muscle fibers is the possibility to show the effect of continuous muscle fibre activity on the parameters of the ECAPs and their spectral characteristics at large radial distance, which is not experimentally accessible. Our results are in agreement with those experimentally obtained. The results from the model prove the role of changes in PV of excitation along the muscle fibres (representing the last link in the complex organized motor system) in the development of fatigue. PMID- 9851023 TI - Analysis of the electroencephalographic activity during the Necker cube reversals by means of the wavelet transform. AB - In previous studies, a perceptual switching related potential was obtained during the observation of a multistable dynamic reversal pattern, where the averaging of the single responses was triggered by subjects pressing a button. The present methodological study aims to increase the signal quality of perceptual switching related potentials considering the dependence of the measurement method on the reaction time of the subject, which may vary significantly during a session, leading to low-amplitude waveform in the averaged event-related-potential (ERP). To overcome this problem in measuring the electrophysiological correlate of an internal event, a pattern selection method based on the wavelet transform (WT) is proposed to choose a subset of single ERPs with more homogenous latencies. Nine subjects observed a Necker cube and were instructed to press the button immediately after perceptual switching. A slow, low-amplitude positive wave with frontocentral amplitude maxima was observed around 250 ms prior to the button press. After the application of a 5 octave WT on single sweeps, the time frequency coefficients obtained in each octave were averaged across trials. The most dominant feature representing the averaged ERP was the delta (0.5-4 Hz) coefficient occurring between 250 and 125 ms before the button press. By averaging the subset of the single sweeps containing this property, a sharpening and significant amplitude increase of the response peak was observed. PMID- 9851024 TI - Studies on the large subunit rRNA genes and their flanking regions of Leuconostocs. AB - The 16S-23S (spacer-1) and 23S-5S (spacer-2) rRNA intergenic spacer regions of Leuconostoc lactis, Leuconostoc mesenteroides, Leuconostoc mesenteroides subsp. dextranicum, and Leuconostoc mesenteroides subsp. cremoris were amplified by polymerase chain reactions and sequenced. The 23S rRNA genes of Leuconostoc lactis, Leuconostoc mesenteroides, and Leuconostoc mesenteroides subsp. dextranicum were also sequenced. The RNase III-like and RNase E processing sites, as well as putative antitermination signals, were identified within the spacer regions. A single tRNA(Ala) gene without the 3'-terminal CCA sequence was found in spacer-1 regions. Secondary structure models are proposed showing interactions between the two spacer regions of leuconostocs. For all strains studied, spacer-1 and spacer-2 were highly conserved and therefore could not be directly used for strain typing. Sequence information on 23S rRNA genes from Leuconostoc species allowed the determination of regions that can be used as targets for diagnostic probes and amplification primers. Secondary structures of variable helical elements of leuconostocs 23S rRNA were constructed and their primary structures were compared with those of several Gram-positive bacteria with low G+C contents. Comparative analysis revealed that restriction analysis of 23S rRNA variable regions appeared to be sufficient for the search for species-specific signatures. Our experimental observations revealed that one form of the rRNA operons was present in leuconostocs. We have also demonstrated the direct linkage between the three species of rRNA genes, which are organized as follows: 5'-16S rRNA-spacer-1 tRNA(Ala)-23S rRNA-spacer-2-5S rRNA-3'. PMID- 9851025 TI - Isolation and characterization of ACC deaminase genes from two different plant growth-promoting rhizobacteria. AB - We have recently proposed that one way that plant growth-promoting rhizobacteria (PGPR) stimulate plant growth is through the activity of the enzyme 1 aminocyclopropane-1-carboxylate (ACC) deaminase, which causes a lowering of plant ethylene levels resulting in longer roots. As part of an effort to understand the role of this enzyme in PGPR, the genes for ACC deaminase from two PGPR, Enterobacter cloacae CAL2 and UW4, have been isolated. These genes are highly homologous to the ACC deaminase genes from Pseudomonas strains 6G5 and F17 and similar to the ACC deaminase gene from Pseudomonas sp. strain ACP. The region downstream (i.e., at the 3'-terminal end) of the strain UW4 ACC deaminase gene has a potential hairpin-like transcription termination site. The regions upstream of the strains UW4 and CAL2 ACC deaminase genes contain putative ribosome-binding sites; however, the promoter sequences have not yet been identified. Southern hybridization experiments suggest that there is a single copy of the ACC deaminase gene in Enterobacter cloacae strains UW4 and CAL2 and that there may be several different types of ACC deaminase genes in different microbes. The cloned ACC deaminase gene can be expressed in Escherichia coli enabling this bacterium to grow on ACC as a sole source of nitrogen and confers upon both Escherichia coli and Pseudomonas spp. strains that are transformed with this gene the ability to promote the elongation of the roots of canola seedlings. PMID- 9851026 TI - Effect of nutrient composition on the in vitro growth of urogenital lactobacilli and uropathogens. AB - Previous clinical studies have shown that nutrients and probiotic agents can alter the composition of the vaginal flora. The present in vitro study has shown that uropathogens have a growth advantage over lactobacilli, but potentially there are natural substances that could be applied vaginally to stimulate lactobacilli growth to the detriment of the pathogens. When chemically defined medium representative of vaginal fluid at pH 5.5 was supplemented with skim milk, it acted as a better substrate for Lactobacillus rhamnosus GR-1 than for uropathogenic bacteria and Candida albicans. Lactobacillus MRS medium, even at pH 4.5, supports the growth of pathogens, but when supplemented with ascorbic acid or EDTA, Lactobacillus growth was significantly higher. When L. rhamnosus GR-1 was coincubated in a combined nutrient composition of vitamins and lactose, it survived better than Escherichia coli and Enterococcus faecalis. These in vitro results provide a basis for testing nutritional supplements to alter the urogenital flora in an attempt to enhance restoration and maintenance of a normal disease-free state. PMID- 9851027 TI - Use of almond mesocarp for production of the entomopathogenic fungus Verticillium lecanii. AB - Almond mesocarp (AM) has been evaluated for production of Verticillium lecanii. Microbial flora of AM has been studied. After ground AM dilution plating, 5.3 x 10(5) +/- 2.6 x 10(4) fungal CFU.g-1 of dry AM were found. Common fungal saprophytes (Rhizopus sp. and Alternaria spp.) were found in more than 80% of the samples. Aspergillus sp. and yeasts were found less commonly. Rhizopus sp., Alternaria spp., and Aspergillus sp. inhibited growth of several V. lecanii; therefore, AM was treated with sterilization agents to eliminate endogenous mycoflora. Small samples (10 g) of AM saturated in distilled water treated with steam (120 degrees C and 100 kPa) were completely sterilized after 15 min. Ground AM incorporated on agar increased the biomass of V. lecanii compared with controls. This suggested AM as suitable substrate for the production of the fungus. In petri dishes, 9.7 x 10(7) +/- 2.9 x 10(7) conidia.g-1 of dry AM were produced after inoculating 10 conidia.g-1 of AM and incubating for 2 weeks. Viability of conidia produced was more than 90%. These conidia (5 x 10(4) conidia per larvae) caused Galleria mellonella mortality, calculated as median lethal time (LT50 5.3 +/- 1.6 days), that was significantly higher (F = 10.93; P < 0.05) than untreated controls (LT50 11.3 +/- 1.1 days). Larger scale tests have to be optimized before mass production. PMID- 9851028 TI - The effect of growth conditions on in vitro adherence, invasion, and NAF expression by Proteus mirabilis 7570. AB - Proteus mirabilis is a common cause of upper urinary tract infections. Fimbriae mediated adherence of this organism to urinary tract epithelium and invasion of host cells are factors thought to be important in its pathogenesis. We have assessed the effect of growth in serum, blood, and urine on the ability of P. mirabilis 7570 to adhere to and invade in vitro the cell line EJ/28, derived from a human urinary tract tumour, and to express nonagglutinating fimbriae (NAF). Proteus mirabilis was capable of adhering to EJ/28 cells to varying degrees depending upon the growth conditions used. It was invasive under all conditions, except when grown in urine, and was found to be particularly so when serum or blood was present in the media. Expression of NAF occurred under all growth conditions examined and was limited only by a decrease in temperature. PMID- 9851029 TI - Tannin-tolerant ruminal bacteria from East African ruminants. AB - Three strains of tannin-tolerant rumen bacteria were isolated from enrichment cultures of rumen microflora of sheep, goat, and antelope and established in medium containing high concentrations of crude tannin extract or tannic acid. These three strains (EAT2, ES3, and EG19), characterized as Selenomonas species, were curved rods, obligately anaerobic, Gram negative, highly motile, and grew in media containing 50 g of tannin extract/L and 50-70 g of tannic acid/L. Strain EAT2 was able to hydrolyze gallic acid, while strains ES3 and EG19 hydrolyzed tannic acid but not gallic acid. All isolates were able to grow in media containing up to 8 g of condensed tannins/L. Growth was very slow when soluble carbohydrate was not added to the medium. PMID- 9851030 TI - Isolation of a class IV chitin synthase gene from a zygomycete fungus, Rhizopus oligosporus. AB - We found the presence of DNA sequence which shows sequence similarity to the class IV chitin synthase gene (CHS3) of Saccharomyces cerevisiae in the genome of 14 Rhizopus species which belong to zygomycetes. We cloned a gene (chs3), which might correspond to one of these homologous sequences, from Rhizopus oligosporus by low stringency plaque hybridization probed with CHS3. The deduced amino acid sequence of this gene showed highest similarity to the class IV chitin synthase of Neurospora crassa (46.7% identity over 1087 amino acids), showing that this gene encodes a class IV chitin synthase. Northern analysis revealed the differential expression pattern of this gene in the asexual life cycle with highest expression in the early stage of asexual spore formation. This is the first report of the isolation and analysis of a class IV chitin synthase gene from zygomycete fungi. PMID- 9851031 TI - Dynamics of in vivo protein aggregation: building inclusion bodies in recombinant bacteria. AB - Time-dependent aggregation of a plasmid-encoded beta-galactosidase fusion protein, VP1LAC, has been carefully monitored during its high-rate synthesis in Escherichia coli. Immediately after recombinant gene induction, the full-length form of the protein steadily accumulates into rapidly growing cytoplasmic inclusion bodies. Their volume increases during at least 5 h at a rate of 0.4 micron3 h-1, while the average density remains constant. Protein VP1LAC accounts for about 90% of the aggregated protein throughout the building process. Minor components, such as DnaK and GroEL chaperones, have been identified in variable, but low concentrations. The homogeneous distribution of inclusion bodies among the cell population and the coexistence of large, still growing bodies with newly appearing aggregates indicate that the aggregation cores are mutually exclusive, this fact being a main determinant of the in vivo dynamics of protein aggregation. PMID- 9851032 TI - Production of cholera toxin B subunit in Lactobacillus. AB - The intracellular expression of the B subunit of cholera toxin (CTB) was first achieved in Lactobacillus paracasei LbTGS1.4 with an expression cassette including the P25 promoter of Streptococcus thermophilus combined with the translation initiation region from the strongly expressed L. pentosus D-lactate dehydrogenase gene (ldhD). Secretion of CTB was next attempted in L. paracasei LbTGS1.4 and L. plantarum NCIMB8826 with four different signal sequences from exported proteins of lactic acid bacteria (Lactococcus lactis Usp45 and PrtP, Enterococcus faecalis unknown protein and S. pyogenes M6 protein). Host-dependent secretion of CTB was clearly observed: whereas none of the secretion cassettes led to detectable CTB in the extracellular fraction of L. paracasei LbTGS1.4, secretion of CTB molecules was clearly achieved with three of the selected signal sequences in L. plantarum NCIMB8826. PMID- 9851033 TI - Malonate decarboxylase of Pseudomonas putida is composed of five subunits. AB - Two different forms of malonate decarboxylase were purified from Pseudomonas putida. The active form was composed of the five different subunits alpha (60 kDa), beta (33 kDa), gamma (28 kDa), delta (13 kDa), and epsilon (30 kDa) and the inactive form was composed of the four subunits lacking the epsilon subunit. The former catalyzed the decarboxylation of malonate to acetate, but the latter could not, although it retained both activities of acetyl-CoA:malonate CoA transferase and malonyl-CoA decarboxylase. The delta subunit of the active form was acylated by the incubation with [2-14C]malonyl-CoA, but the delta subunit of the inactive form was not labeled. From the above results and the N-terminal amino acid sequence analysis, it was concluded that the epsilon subunit was an essential subunit to function as malonyl-CoA:ACP transacylase, which was an indispensable component of the enzyme for the cyclic decarboxylation of malonate. PMID- 9851034 TI - Legionella pneumophila kills human phagocytes but not protozoan host cells by inducing apoptotic cell death. AB - Legionella pneumophila is a facultative intracellular parasite able to replicate within and to kill a variety of eukaryotic cells. One possible killing mechanism is the induction of programmed cell death. Based on electron microscopy and flow cytometry studies using the phosphatidylserine binding protein annexin V, we could demonstrate that L. pneumophila is able to induce apoptosis in human monocytes which was clearly dependent on the multiplicity of infection, the time postinfection and the intracellular location of the bacteria. Furthermore, it became evident that Legionella-induced apoptosis does not require the TNF-alpha mediated signal-transduction pathway. By studying infection in Acanthamoeba castellanii, we found that L. pneumophila is not able to induce programmed cell death in their natural host cells indicating that different mechanisms are responsible for host cell killing in protozoan and human cells. PMID- 9851035 TI - Sequence, identification and effect on conjugation of the rmoA gene of plasmid R100-1. AB - The rmoA gene was recently identified from two partially overlapping sequences corresponding to a region close to the end of the tra operon of plasmid R100. Its putative amino acid sequence showed strong homology to the Hha protein of Escherichia coli and YmoA protein of Yersinia enterocolitica, which are modulators of gene expression in response to environmental stimuli. We have cloned the rmoA gene from plasmid R100-1 in pUC19 and obtained the complete nucleotide sequence, which was previously published only partially and may have contained some mistakes. The rmoA gene product has been identified in radiolabelled minicells as a protein of the predicted molecular mass. The wild type rmoA gene of plasmid R100-1 has been mutated by gene replacement and its effect on the efficiency of conjugation has been analysed. When grown in LB medium, cells harbouring R100-1 plasmid with a disrupted copy of rmoA showed a five-fold increase in conjugation frequency compared to cells harbouring R100-1 plasmid with the wild-type rmoA gene, grown in the same conditions. When cells were grown in NaCl-free LB medium they showed a 50-fold increase in conjugation frequency. PMID- 9851036 TI - Molecular and ultrastructural characterisation of EspA from different enteropathogenic Escherichia coli serotypes. AB - Enteropathogenic Escherichia coli (EPEC) encode a type III secretion system located on a pathogenicity island known as the locus for enterocyte effacement. Four proteins are known to be exported by this type III secretion system--EspA, EspB and EspD required for subversion of host cell signal transduction pathways and a translocated intimin receptor protein (Tir) required for intimin-mediated intimate attachment and attaching and effacing lesion formation. The espA gene is located within the locus for enterocyte effacement and the EspA polypeptide from the prototype EPEC strain E2348/69 (O127:H6) has recently been shown to be a component of a filamentous structure involved in bacteria-host cell interaction and locus for enterocyte effacement-encoded protein translocation involved in attaching and effacing lesion formation. In this study we have extended our investigation of EspA to strains belonging to other classical EPEC serotypes. DNA sequencing demonstrated that the espA gene from the different EPEC strains share at least 65% DNA identity. In addition, we detected morphologically and antigenically similar EspA filaments in all but one of the bacterial strains examined including recombinant, non-pathogenic E. coli expressing espA from a cloned locus for enterocyte effacement region (HB101(pCVD462)). PMID- 9851037 TI - Maltose metabolism of Lactobacillus sanfranciscensis: cloning and heterologous expression of the key enzymes, maltose phosphorylase and phosphoglucomutase. AB - The maltose degradation operon containing genes encoding maltose phosphorylase mapA and phosphoglucomutase pgmA from Lactobacillus sanfranciscensis DSM20451T were cloned and expressed in Escherichia coli. These genes represent the first genetic data available for this species beyond taxonomic classification. MapA encodes a 754-amino acid polypeptide representing maltose phosphorylase, MapA, with a calculated molecular mass of 85.7 kDa. Comparative sequence analysis showed that mapA is of a new type distinct from other alpha-glucosidase genes sequenced so far. Putatively, pyridoxal 5'-phosphate is required as cofactor. The deduced amino acid sequence of pgmA shows an overall similarity of 39% to the phosphoglucomutase of Lactococcus lactis. pgmA is separated by a single nucleotide from the preceding mapA gene indicating effective translation by translational coupling. Upon subcloning mapA was heterologously expressed in E. coli. Additionally, upstream of the maltose-degrading operon ORF1 and ORF2 are located in the opposite direction. These genes show homology to fabZ and accB from E. coli and Bacillus subtilis, respectively, both involved in fatty acids biosynthesis. PMID- 9851038 TI - Hydrocarbon uptake by Streptomyces. AB - Oligocarbophilic Streptomyces strains capable of hydrocarbon uptake and utilization were isolated from the polluted desert of Kuwait and used in this study. Transmission electron-microscopy of hyphae revealed that they become enriched with large less electron dense areas in the cytoplasm, when biomass samples were incubated with alkanes. The Streptomyces isolate could utilize n hexadecane as sole carbon and energy source and their fatty acid content showed an increase in the fatty acids with chain length equivalent to those of the alkane substrates. Fluorescence measurements of diphenylhexatriene dissolved in the representative alkane, n-hexadecane, showed that the kinetics of hydrocarbon uptake are quite different in hydrocarbon-utilizer compared with non-utilizer Streptomyces strain. Microviscosity of the cellular membrane of the utilizer strain was also different from that of the non-utilizer control strain Streptomyces griseus after incubation in the presence of n-hexadecane. Very likely the hydrocarbon utilizer transported these compounds more efficiently across their membranes and accumulated them as inclusions in the cytoplasm. PMID- 9851039 TI - In vivo apoptosis of hepatocytes in guinea pigs infected with Leptospira interrogans serovar icterohaemorrhagiae. AB - To investigate the contribution of the previously demonstrated in vitro apoptosis to the pathogenesis of leptospirosis, guinea pigs were infected with Leptospira interrogans serovar icterohaemorrhagiae strain Verdun and sequentially killed to collect target organs involved in the natural history of the disease (liver, kidneys, lungs, spleen and heart). The combination of histopathological procedures and a specific TUNEL assay showed a significant Leptospira-induced programmed cell death of hepatocytes with a peak at 48 h post inoculation. Hepatocyte nuclei showed morphological changes including fragmented and condensed nuclei. This phenomenon occurred early in the course of the disease at a time where infecting leptospires were present at a low density between the liver parenchyma cells. PMID- 9851040 TI - Gene encoding a replication initiator protein and replication origin of conjugative plasmid pSA1.1 of Streptomyces cyaneus ATCC 14921. AB - pSA1.1 is a 9.1-kb multicopy plasmid originally isolated from Streptomyces cyaneus (formerly S. azureus) ATCC 14921. This plasmid accumulates single stranded DNA in S. lividans and is therefore considered to replicate by a rolling circle replication. In the present work, the rep gene encoding the replication initiator protein and the replication origin ori of pSA1.1 were determined. The rep and ori are located on separate regions. The Rep protein of pSA1.1 belongs to superfamily I which includes A proteins of phages. Nucleotide sequence of the surrounding putative nicking site of pSA1.1 shows good agreement with those of the pC194 group plasmids and phages. The direction of replication was also determined. PMID- 9851041 TI - Molybdate-dependent transcription of hyc and nar operons of Escherichia coli requires MoeA protein and ModE-molybdate. AB - In Escherichia coli, ModE-molybdate, a repressor of modABCD operon (molybdate transport), was previously shown to be an additional transcriptional activator of hyc operon (formate hydrogenlyase) and narGHJI operon (respiratory nitrate reductase). However, in a modE mutant, both operons were expressed at about 50% of the wild-type level in a molybdate-dependent manner. This ModE-independent, molybdate-dependent, expression of hyc, narG and narK operons required MoeA protein. An E. coli modE, moeA double mutant failed to produce formate hydrogenlyase or respiratory nitrate reductase activity irrespective of the growth medium. Tungstate substituted for molybdate in the activation of transcription of hyc and nar operons by ModE could not replace molybdate for MoeA dependent expression. It is proposed that the MoeA-catalyzed product, an activated form of molybdate, interacts with a transcriptional activator/regulator other than ModE and regulates hyc and nar operons. PMID- 9851042 TI - Analysis of the Mycobacterium bovis hsp60 promoter activity in recombinant Mycobacterium avium. AB - A clinical isolate of Mycobacterium avium was transformed with a new shuttle plasmid containing the Escherichia coli beta-galactosidase reporter gene under the control of the Mycobacterium bovis bacillus Calmette-Guerin (BCG) hsp60 promoter. beta-Galactosidase activity was assayed spectrophotometrically in bacterial homogenates of the recombinant strain (M. avium::lacZ) and used for quantification of the hsp60 promoter strength in different conditions of extra- and intracellular growth. Very low levels of beta-galactosidase were recorded during the exponential phase of in vitro growth, while they increased progressively during the late exponential and stationary phases. A significant increase in enzyme activity was also induced in exponentially growing cells by shifting the incubation temperature from 37 to 45 degrees C, but not from 37 to 42 degrees C nor from 30 to 42 degrees C. No induction of the promoter was observed by adding hydrogen peroxide to the cultures. Finally, beta-galactosidase levels were quantified during growth of M. avium::lacZ in murine macrophages. Soon after phagocytosis and, to a lesser extent at 1, 5 and 7 days after infection, increased levels of bacterial beta-galactosidase were observed indicating an increment in transcriptional activity of hsp60 promoter both at early phases of infection and during the course of intracellular growth. PMID- 9851043 TI - Importance of pre-enrichment media for isolation of Salmonella spp. from swine and poultry. AB - The performance of two new (1-day) culture methods, Salmonella Enrichment Broth (SEB) and Revive, and an alternative pre-enrichment broth, designated Universal pre-enrichment broth (UB), was compared to the internationally accepted buffered peptone water (BPW). The study was directed towards detection of Salmonella in 100 faecal samples from porcine and 100 neck-skin samples from poultry. The sensitivity (number of positive cases per method among all the positive cases) of the conventional pre-enrichment in BPW was found to be 0.77 for swine and 0.66 for poultry samples, while a combination of the BPW method with parallel pre enrichment of the same sample in UB resulted in high sensitivity for swine (0.92) and poultry (0.95) samples. A 2-h pre-enrichment in the non-selective Revive, followed by overnight enrichment in selective broth, resulted in a low sensitivity, particularly for the neck-skin samples (0.16, P = 0.001). The SEB method in the porcine samples resulted in a sensitivity (0.71) comparable to the standard method (P = 0.31). In conclusion, additional pre-enrichment of samples in UB may substantially increase the culture sensitivity. During routine screening of large numbers of samples, it may be advantageous to use SEB rather than standard culturing. PMID- 9851044 TI - Nature of Candida albicans-derived carbohydrate antigen recognized by a monoclonal antibody in patient sera and distribution over Candida species. AB - The minimal epitope of an anti-Candida albicans mannan monoclonal antibody (MAb) EB-CA1, used to detect mannanemia in patient sera, was determined, MAb EB-CA1 exhibited reactivity with oligomannosides released from the mannan acid stable domain, converted into neoglycolipids (NGLs) and coated onto ELISA plates. Reactivity occurred with mannopentaose and higher oligomers, whereas mannotriose and mannotetraose were unreactive. MAb EB-CA1 binding to mannan acid stable mannopentaose NGL displayed a dose dependent and saturable specific reactivity curve whereas there was a complete absence of binding, even at high concentrations, with NGLs constructed from the beta-1,2-linked mannopentaose derived from the mannan acid labile fraction. MAb EB-CA1 binding to acid stable mannopentaose NGL was inhibited by the homologous oligomannoside but not by mannotriose and mannotetraose. NMR analysis showed that mannotriose and mannotetraose contained exclusively alpha-1,2-linked D-mannopyranose units and that mannopentaose was a mixture of a mannopentaose alpha-1,2-linked and an isomer in which the fifth mannose was alpha-1,6-linked to the reducing unit of manno-alpha-1,2 tetraose. Western blot analysis has shown that MAb EB-CA1 epitope was expressed on a wide range of C. albicans manno-glycoconjugate as well as on manno-glycoconjugates of other pathogenic species of the genus Candida, viz. C. tropicalis, C. glabrata, C. parapsilosis and C. krusei. PMID- 9851045 TI - Target cell specificity of the Pasteurella haemolytica leukotoxin is unaffected by the nature of the fatty-acyl group used to activate the toxin in vitro. AB - The leukotoxin (LktA) of Pasteurella haemolytica is active only against cells of ruminant origin. It is synthesised as an inactive protoxin encoded by the lktA gene and post-translationally modified to the active toxin by the product of the lktC gene. The LktA and LktC proteins were expressed separately in Escherichia coli and partially purified. Active LktA was produced in vitro in the presence of LktC and acyl-acyl carrier protein (ACP) charged separately in vitro with a fatty acyl group. The toxic activity and target cell specificity of LktA and adenylate cyclase toxin (CyaA), a toxin active against a wide variety of mammalian cells, were investigated after activation with ACP charged with different fatty acids. Palmitoyl-ACP produced the most active toxin in both cases and, although other fatty acids were also effective, the fatty acid preference was the same for the in vitro activation of both toxins. Activated LktA remained ruminant cell specific whichever acyl group was used to acylate the A protoxin. PMID- 9851046 TI - High levels of transcription factor RpoS (sigma S) in mviA mutants negatively affect 1,2-propanediol-dependent transcription of the cob/pdu regulon of Salmonella typhimurium LT2. AB - Expression of the cobalamin biosynthetic (cob) and 1,2-propanediol utilization (cob/pdu) regulon of Salmonella typhimurium LT2 is controlled at the transcriptional level by global and specific regulatory proteins. In this paper we show that mutations in the mviA gene negatively affect cob/pdu transcription in response to 1,2-propanediol in the environment. The effects of mviA mutations were consistent with its role in the regulation of RpoS levels in the cell. Null mutations in rpoS eliminated the negative effect of mviA mutations on cob/pdu transcription, and restored growth on succinate, propionate and 1,2-propanediol. In addition, mviA mutants were deficient in the utilization of succinate, propionate and 1,2-propanediol as carbon and energy sources. PMID- 9851047 TI - Identification and sequence analysis of Treponema pallidum tprJ, a member of a polymorphic multigene family. AB - TnphoA mutagenesis was used to identify genes encoding exported proteins in a genomic DNA library of Treponema pallidum, the syphilis agent. The nucleotide sequence of an open reading frame (tprJ) that encodes a 755-amino acid protein with a predicted molecular mass of 81.1 kDa was determined. The deduced amino acid sequence of TprJ has homology to the major surface protein of Treponema denticola, a periodontal pathogen. Southern hybridization and genomic DNA sequence analysis indicate that tprJ is a member of a polymorphic multigene family. RT-PCR data showed that tprJ is expressed in treponemes during syphilitic infection. A putative tprJ gene was sequenced from T. pertenue, the closely related yaws agent. The deduced amino acid sequence of T. pertenue TprJ is 87.3% identical to that of T. pallidum TprJ. This is the first report of significant sequence differences within homologous genes of T. pallidum and T. pertenue. PMID- 9851048 TI - The leader sequence of the Escherichia coli lysC gene is involved in the regulation of LysC synthesis. AB - In Escherichia coli and Bacillus subtilis, long leader sequences are found upstream of the lysC coding sequences which encode lysine-sensitive aspartokinase. Highly conserved regions exist between these sequences. Mutations leading to constitutive expression of the E. coli lysC gene have been localised within these conserved regions, indicating that they participate in the lysine mediated repression mechanism of lysC expression. PMID- 9851049 TI - Both the full-length and the N-terminal domain of the meningococcal transferrin binding protein B discriminate between human iron-loaded and apo-transferrin. AB - We have readdressed the ability of the transferrin-binding protein B (TbpB) from Neisseria meningitidis to discriminate between the iron-loaded and the iron-free human transferrin (hTf) by using the BIAcore technology, a powerful experimental technique for the observation of direct interactions between a receptor and its ligands, without the use of labels. Recombinant full-length TbpB from five N. meningitidis strains were produced and purified from Escherichia coli as fusion proteins. They showed a preference for the binding to iron-loaded hTf. As for the full-length molecule, we have demonstrated that the minimal N-terminal hTf binding domain of meningococcal TbpB from B16B6 and M982 strains was able to discriminate between both hTf forms. PMID- 9851050 TI - Construction of improved plasmid vectors for promoter characterization in Pseudomonas aeruginosa and other gram-negative bacteria. AB - We report the construction of two broad host range promoter-probe plasmid vectors for rapid analysis of promoters in Gram-negative bacteria. The new vectors, pME4507 and pME4510, carry carbenicillin and gentamycin resistance genes, respectively, and are small sized (4 kb) with a flexible multiple cloning site to facilitate directional cloning of putative promoter elements. The vectors allow rapid plate-based screening for promoter activities, using beta-galactosidase as the reporter enzyme. In the absence of an inserted promoter fragment, they display very low background activity, making them a useful tool for analysis of low expression level promoters. PMID- 9851051 TI - Genetic transformation of germinated conidia of the thermophilic fungus Humicola grisea var. thermoidea to hygromycin B resistance. AB - Germinated conidia of the thermophilic fungus Humicola grisea var. thermoidea were transformed to hygromycin B resistance using the plasmid pAN7.1. Transformation was achieved using lithium acetate treatment or electroporation. The efficiency of transformation was up to 32 and 25 transformants per microgram of plasmid DNA with the two methods, respectively. Transformants obtained by the lithium acetate method were more stable and showed a high copy number of the hph gene integrated into their genome. The other transformants, from the electroporation procedure, were stable, but unable to grow in the presence of high levels of hygromycin, and detection of the hph gene was only possible by polymerase chain reaction analysis. PMID- 9851054 TI - Structure determines function of the retina, a neural center. 4. The 'duplex' nature of vision. AB - Part 4 reveals how the differences in the quality of vision at high and low illumination are explained by synaptic interaction between the neurons in the outer plexiform layer changing with illumination. Analysis of data available in the literature revealed that rod vision is active over the entire range of light intensities generating visual responses. It is revealed that the input from the retina to brain centers involves information regarding a black and white image based on rod vision, which is improved with respect to spatial brightness contrast and darkness contrast by cone interference. Cones add color to the image, further increasing image contrast. The duplicity theory is shown to be based on erratic assumptions regarding how rods and cones function. The theory lacks an experimental basis and is not required to explain differences in the quality of the perceived images at high and low illumination. PMID- 9851055 TI - Diabetes-induced structural changes of venous and arterial endothelium and smooth muscle cells. AB - The structural alterations of endothelium and smooth muscle cells of the hind limb and heart veins and arteries were investigated in Golden Syrian hamsters subjected to streptozotocin induced diabetes. Animals were examined at 5, 10, and 15 weeks after induction of diabetes. At each time point body weight and plasma glucose concentrations were recorded. Anesthetised animals were washed out of blood, fixed in situ, and the femoral vein and artery, saphenous vein and artery, and heart veins and coronaries were dissected out, and processed for electron microscopical examination. Anionic sites of the endothelial plasmalemma were visualized by in situ perfusion of cationized ferritin. The endothelial localization of von Willebrand factor was carried out by immunocytochemistry. The results showed that induction of experimental diabetes generated morphological changes of the endothelium and smooth muscle cells of both hind limb and heart vessels. The common alterations developed in endothelial cells of venous and arterial origin consisted in: 1) the development of a secretory phenotype, enriched in biosynthetic and degradative organelles; 2) the abundance of cytoskeletal elements, especially intermediary filaments; 3) the increase in number of fused plasmalemmal vesicles and transendothelial channels, and 4) the hyperplasia of the basal lamina. In contradistinction to the arterial endothelium, the peculiarities of the venous endothelium in the diabetic hamsters examined were: 1) the uniform distribution of the anionic sites exposed on the luminal plasma-lemma (as in normal animals), and 2) the increased number of copies of Weibel-Palade bodies (up to 13 copies per endothelial cell in the hind limb). Von Willebrand factor was immunodetected in Weibel-Palade bodies, Golgi cisternae and some vesicles of normal and diabetic hamsters. With time, and especially pronounced at 15 weeks of diabetes, the smooth muscle cells of veins and arteries examined exhibited a characteristic secretory phenotype, and were surrounded by a reticulated basal lamina and a hyperplasic extracellular matrix (especially pronounced in arteries). These data indicate that diabetes affects both heart and hind limb veins and arteries, producing structural changes of the endothelium and smooth muscle cells which may account, at least in part, for the specific vascular complications. PMID- 9851052 TI - Stress tolerance in a yeast sterol auxotroph: role of ergosterol, heat shock proteins and trehalose. AB - The role of ergosterol in yeast stress tolerance, together with heat shock proteins (hsps) and trehalose, was examined in a sterol auxotrophic mutant of Saccharomyces cerevisiae. Ergosterol levels paralleled viability data, with cells containing higher levels of the sterol exhibiting greater tolerances to heat and ethanol. Although the mutant synthesised hsps and accumulated trehalose upon heat shock to the same levels as the wild-type cells, these parameters did not relate to stress tolerance. These results indicate that the role of ergosterol in stress tolerance is independent of hsps or trehalose. PMID- 9851056 TI - Spermiogenesis in phytophagous bug (Hemiptera, Pentatomidae): an ultrastructural study. AB - Ultrastructural and immunocytochemical studies were carried out on spermiogenesis in two species of phytophagous bug, Acrosternum aseadum and Nezara viridula. The nucleus development involved changes in the shape and in the degree of chromatin condensation, with specific aggregation patterns of DNA-histone complex occurring during this process. The acrosome showed a three-layered and the acrosomal content a paracrystalline arrangement. The axoneme presented a 9 + 9 + 2 microtubule pattern and bridges occurred between axonemal microtubules 1 and 5, and mitochondrial derivatives. Two paracrystalline structures embedded in amorphous regions were observed in the mitochondrial derivatives. The use of the negative staining technique shows a zig-zag profile in the mitochondria due to infolding to the cristae, regularly spaced with approximately 40 nm. An electron dense rod was observed near the centriolar adjunct; it presented labelling for tubulin suggesting that this structure may be involved in the microtubule organization during spermiogenesis of these insects. PMID- 9851057 TI - The sinus endothelial cell architecture in the mouse lymph node. Structural peculiarities and close correlation with the fibroblastic reticular cells. AB - The sinus endothelial cell architecture has been studied in the mouse lymph node by light and electron microscopy after tissue fixation with an osmium-zinc iodide mixture. This histochemical method allowed visualization of sinus endothelial cells in the medullary and, partly, in the intermediate sinuses. The osmium-zinc iodide reaction also demonstrated fibroblastic reticular cells, which represent the main stromal cell population in the lymph node and form the skeletal framework of the organ as well as the adventitia of sinuses. In the mouse lymph nodes, sinuses were lined by a simple layer of sinus endothelial cells whose integrity and continuity showed interruptions of various extension. In areas devoid of complete sinus endothelial cell lining, fibroblastic reticular cells located in the nearby parenchyma were able to reach the border of the sinus, being thus in direct contact with the lumen content. Dendritic sinus endothelial cells as well as intermediate forms between sinus endothelial cells and fibroblastic reticular cells could be observed. The close structural characteristics that sinus endothelial cells have in common with fibroblastic reticular cells and the finding of transitional cells with intermediate morphology between sinus endothelial cells and fibroblastic reticular cells, suggest a possible origin of sinus endothelial cells by migration and differentiation of fibroblastic reticular cells located in the sinus adventitia. PMID- 9851058 TI - T (Thomsen-Friedenreich) and Tn epitope location and their spatial relations to adhesion plaques on human breast carcinoma cells: immunogold-silver staining studies at scanning electron microscopic level. AB - T and Tn are pancarcinoma epitopes (EPs) which can be immunodetected in about 90% of adenocarcinomas (CAs). To study the location of T and Tn EPs and their relations to adhesion plaques on CA cells, immunogold-silver staining method was employed at scanning electron microscope (SEM) level. Human breast CA cells grown on coverslips were fixed in paraformaldehyde and glutaraldehyde, reacted with cocktails of monoclonal antibodies against T or Tn EPs, followed by incubation with 10 nm gold conjugated goat anti-mouse immuno-globulins. The positive gold particle labelling was amplified with silver enhancement solution, and the specimens were then routinely critical point dried, sputter-coated with palladium and observed under SEM. The studies show that T and Tn EPs are not randomly distributed on CA cell surface; they are aggregated at the adhesion plaque area. These results confirm that T and Tn EPs play roles in CA cell adhesion, and suggest that they may represent the initial points of contact by immuno-effectors and therefore can be utilized in immunointervention and anti-adhesion therapy against CA. PMID- 9851059 TI - Storage of trachea for X-ray microanalytical studies of tracheal submucosal glands. AB - Pig trachea is a good model system to study airway fluid transport with relevance to human diseases such as cystic fibrosis, since both in pig and in man, the submucosal glands play a quantitatively important role in fluid secretion. Pig trachea can be easily obtained from the local slaughterhouse. In the present study, we investigated the procedure to transport the tissue from the slaughterhouse to the laboratory where physiological experiments can be carried out. Transport of the tissue in a solution with high K+ content (resembling the intracellular environment) maintained the morphology of the glands and a high intracellular K/Na ratio. Physiological experiments can be carried out in solutions with a high Na+ concentration resembling the extracellular environment. The glands secrete chloride on stimulation with alpha-adrenergic, beta adrenergic, cholinergic and peptidergic agonists. Transport and temporary storage of tissue in high K+ solutions expands the scope of possible X-ray microanalytical studies in the field of pathophysiology. PMID- 9851060 TI - Effects of nicotine administration on elemental concentrations in mouse granulosa cells, maturing oocytes and oviduct epithelium studied by X-ray microanalysis. AB - A normal maturation of the oocytes is dependent upon, among other things, normally functioning granulosa and corona radiata cells. Analyses performed during human in vitro fertilization programs have revealed that, in smokers, ovarian functions are affected and that smokers have a decreased fertilization rate. Further, animal studies have indicated that nicotine can reach the genital tractus, and that nicotine administration interferes with oocyte maturation, fertilization and early pregnancy. We applied X-ray microanalysis to monitor whether nicotine administration changed the ionic balance of cells in the reproductive tract (granulosa cells, oocytes and oviduct epithelial cells). The animals were given nicotine in the drinking water at a concentration of 108 mumol/l. After 15 days the animals were superovulated, ovaries and oviducts were frozen, and thick cryosections were prepared for energy-dispersive X-ray microanalysis. In the granulosa cells, the concentrations of Na and Cl increased after nicotine treatment, while the K concentrations decreased resulting in an increased Na/K ratio. The treated oocytes had a higher K concentration and a decreased Na/K ratio compared to the controls. In the epithelial cells of the oviduct, the concentrations of Na and K decreased after nicotine treatment without any changes in the Na/K ratio. Thus, heavy nicotine administration to mice causes significant changes in the ionic composition of the granulosa cells, the ovarian oocytes and the oviduct epithelium. PMID- 9851062 TI - Interstitial tissue in the testis of the giant rat, Cricetomys gambianus. AB - Besides Leydig cells macrophages form considerable part of the interstitial tissue in the giant rat, Cricetomys gambianus. Leydig cells contain bundles of usually six to twelve parallel tubes with walls of longitudinal layers alternating with circular layers of microfilaments. Polyribosomes, both free and membrane bound, are concentrated around or inside the tubes. The tubes may enclose mitochondria and microperoxisomes as well. The light cytosol of the macrophages is dotted with electron-dense 5-nm particles which might be ferritin. Specific contacts exist between Leydig cells and macrophages indicating a functional interdependence between these two cell types. PMID- 9851061 TI - Association in the same patient of autosomal dominant progressive external ophthalmoplegia with multiple mtDNA deletions and X-linked ichthyosis: clinical, biochemical, histological, submicroscopic and molecular genetic study. AB - Autosomal dominant chronic progressive external ophthalmoplegia (AdPEO) is a muscle mitochondrial disorder due to multiple large scale rearrangements of the mitochondrial DNA. This disorder is probably due to a nuclear defect which causes genetic instability or an impairment in the replication of mitochondrial DNA. X linked ichthyosis (XLI) is a skin disorder caused by a deletion in the steroid sulphatase gene. Here we report the clinical, biochemical, morphologic and molecular genetic findings in a patient affected by both AdPEO, inherited by the father, and steroid-sulphatase-deficiency, inherited by the mother. The association in the same patient of the two inherited diseases is merely casual and does not seem to influence the phenotypic expression of the two diseases. PMID- 9851063 TI - Three-dimensional and ultrastructural aspects of the lymphatic vascularization of the vermiform appendix. AB - The aim of the study is to reveal the three-dimensional distribution and ultrastructure of the peripheral absorbing lymphatic vessels of the vermiform appendix, since the gut-associated lymphoid tissue is necessary to the immune responses to the enteric antigens. Corrosion casts showed the beginning of the lymphatic vascularization at the tunica mucosa, which lacks intestinal villi, through a tight, delicate lymphatic network. This network drains the lymph by peculiar straight vessels, distributed in the mucosal beams that separate the adjacent follicle domes, in the fine network of the upper portion of the lymphatic basket, surrounding the lateral walls of the basal and medium portions of each lymphoid follicle. This network, which is made of large caliber vessels that are not dilated like sinuses, continues through small vessels into the large dome-like vessels of the submucosa, which in turn by way of the lymphatic vessels of the muscular tunica, drain into the subserous precollector valved lymphatic vessels that flow into the pre-lymph node collectors. We underlined that the particular fluidity of Neoprene latex and the direct injection method, when compared with other substances and injection methods, provided us with exceptionally clear and precise three-dimensional plastic images of the absorbing lymphatic vessels. Moreover, these images extraordinarily illustrated the preservation of the absorbing lymphatic spatial relationships with blood vessels. Ultrastructural features and three-dimensional models of ultrathin serial sections of the absorbing peripheral lymphatic vessels showed a continuous endothelial wall lacking basal lamina, as well as open junctions between adjacent cells. Moreover, we observed the presence of numerous lymphocytes, together with intense transendothelial migratory activity that occurs through intraendothelial channel formations, dynamic entities, at absorbing lymphatic vessels of the peri interfollicular lymphoid tissue. Also, we saw that the germinal center, as well as the lymphoid follicle dome, lacked lymphatic absorbing vessels. In addition, many postcapillary high endothelial venules (HEV) were observed with lymphocyte migration into the extravasal compartment. Furthermore, we maintain that the absorbing peripheral lymphatic vessels (ALPA) of the tunica mucosa play an important role in liquid drainage. For the peri-interfollicular vessels, we hypothesize a potential migratory and a reserve capacity for lymphocytes, as well as a conduction activity for the muscular tunica and submucosa vessels. PMID- 9851064 TI - High plains disease of corn and wheat: ultrastructural and serological aspects. AB - Virus-like particles with unique size and morphology were consistently associated with a new eriophyid mite-borne disease of maize and wheat, the high plains disease. In cells of symptomatic leaves, double membrane-bound particles (DMPs), quasi-spherical structures 120-200 nm in diameter, were present throughout the cytoplasm in association with electron-dense amorphous inclusions. No DMPs and inclusions were observed in symptomless plants. The DMPs were morphologically indistinguishable from those associated with eriophyid mite-borne diseases of uncertain etiology: fig mosaic, rose rosette, yellow ringspot of redbud, thistle mosaic, wheat spot chlorosis and wheat spot mosaic diseases. The DMPs and associated viroplasm-like inclusions in maize and wheat were specifically immunogold labeled in situ with an antiserum to the 32 kDa protein associated specifically with the high plains disease. Thread-like structures, present in the purified preparations from diseased maize were also immunogold labeled with the antiserum. It is suggested that the thread-like structures are derived from the DMPs. In many cells of symptomatic maize and wheat samples, DMPs occurred together with flexuous rod-shaped particles and cylindrical inclusions of wheat streak mosaic potyvirus (WSMV), indicating that these cells are infected doubly with WSMV and the agent represented by the DMPs. PMID- 9851065 TI - Localization of lamins in mammalian spermatozoa. AB - Little information is available concerning lamins in the nucleus of germinal cells. In this paper we briefly describe and compare the organization of A- and B type lamins in several mammalian spermatozoa. Nuclear lamin B is localized primarily in the postacrosomal sheath of human, bull and rabbit spermatozoa; lamin A/C is a major component of the equatorial segment in most of mammalian sperm, with the exception of rodents. In mouse sperm, devoid of equatorial segment, only lamin B appears to be expressed. The same happens in human pathological spermatozoa in which the equatorial segment is altered or absent. PMID- 9851066 TI - Atypical cilia in the oviductal epithelium of healthy dogs. AB - Specimens of the uterine tube (ampulla) were obtained from seven healthy, ovario hysterectomized dogs. Ultrastructurally, a total of 35,000 cilia were examined. Compound cilia ranged from 0.0 to 0.4%; both intracytoplasmic and swollen cilia ranged from 0.1% to 0.4%. The microtubular pattern was studied in 3,500 cross sectioned cilia and an abnormal pattern was found in 2-5%. Similarly to the other animal species, abnormalities involving the peripheral microtubules were the prevailing defect. An electron-dense plug into the lumen was seen in 1-3% of the basal bodies; occasionally an abnormal spatial configuration of them was also observed. The incidence of abnormal cilia hence is lower than found in the tracheae. PMID- 9851067 TI - Prevalence of and factors related to pneumoconiosis among foundry workers in central Taiwan. AB - The objective of this study was to investigate the prevalence of and factors related to pneumoconiosis in foundry workers. Seven hundred and eighteen workers from 50 foundries in central Taiwan were interviewed using a constructed questionnaire. A full-sized PA chest radiograph was used to diagnose pneumoconiosis, according to ILO criteria. Overall, pneumoconiosis was found in 7.5% of the workers. The highest prevalence was found among furnace workers (15.9%) and molding workers (8.40%). All foundry workers except those in administrative positions had a significantly increased risk of developing pneumoconiosis. Using a multiple logistic regression, compared to administrative workers, furnace workers had the highest risk (10.63 times), followed by post treatment workers (6.63 times), and molding workers (5.41 times). In conclusion, the prevalence of pneumoconiosis was significantly related to high concentrations of dust, especially with a high proportion of free silica, however, smoking and length of exposure were also contributing factors. PMID- 9851069 TI - A survey of present levels of radiocesium in Swedish pulp mill liquors and the implications for wood radiocesium transfer factors. AB - Assessments of the impact from radioactive fallout on the pulp industry and the accumulation processes for radionuclides in such industries, using wood and water as primary raw materials, have been conducted in previous studies. Hitherto, there is a general lack of data on present levels of radioactivity in pulp mill liquors apart from the few mills studied by Nilsson (1992); Ravila and Holm (1992); Manjon et al. (1996); Krosshavn et al. (1997). A survey was therefore initiated to describe and compare the present activity levels in pulp mill liquors from various mill locations in Sweden. The 137Cs activity in pulp mill liquors were compared with the mill location and the deposition pattern of the Chernobyl fallout and nuclear weapons fallout. The large input of wood (approx. 2500 m3 day-1) to an average Nordic Kraft mill and the relatively long-term retention time for radiocesium in the Kraft mill recovery cycle enables representative sampling of substances directly related to the activity concentration in wood. PMID- 9851068 TI - Relative risks of transformation products of pesticides for aquatic ecosystems. AB - In this study, the availability of physico-chemical and ecotoxicological information on 78 transformation products for 20 regularly used pesticides in the open literature is evaluated. Based on this information, it is attempted to predict the relative risk for the aquatic environment of each transformation product in comparison to its parent pesticide. It is concluded that for the 78 transformation products selected, the data set on physico-chemical and ecotoxicological behaviour is not very large. Measured log Kow values and other physico-chemical properties are known for only 30-40% of the selected transformation products. The overall reliability of the collected physico chemical values is considered to be moderate to slight, while for the ecotoxicological data set, reliability is considered to be sufficient. In many cases, there is a need for more information especially on the persistency and no observed-effect concentrations of the pesticide's transformation products. In general, over 50% of the transformation products of triazines, carbamates and phenoxypropionic acids pose, in theory, a similar to higher risk than their parent pesticide, while over 50% of the transformation products of synthetic pyrethroids, organophosporous pesticides and dithiocarbamates probably pose less risk. High risk was expected for products with high accumulation or persistency in sediment and/or high toxicity together with considerable bioaccumulation (potential) or relatively high concentrations or persistency in water. A generalization of the joint features that caused an increased risk for ecosystems could not be made for most pesticide classes. Exceptions are the synthetic pyrethroids, for which transformation products with a similar or even higher log Kow than the parent pesticide caused an increased potential risk, while for the carbamates the presence of the carbamate group in the transformation product was the joint characteristic of chemicals with predicted increased risk. For three transformation products, monitoring data based on concentrations measured in surface water in The Netherlands were compared with maximum permissible concentrations. This comparison indicated that two of these compounds pose a potential risk of adverse effects in the field situation in Dutch aquatic ecosystems. For all other transformation products, the potential risk in the field situation could not be established because of the absence of monitoring data. PMID- 9851070 TI - Distribution of cesium-137 in Japanese forest soils: correlation with the contents of organic carbon. AB - The spatial and vertical distributions of 137Cs in surface soils were surveyed and analyzed then correlated with the contents of organic carbon in the hinoki (Chamaecyparis obtusa Sieb. et Zucc.) plantation forest and secondary forest dominated by red pine (Pinus densiflora Sieb. et Zucc.) in Japan. The spatial variation of 137Cs activity was observed in the surface soil around the red pine. The average activity of 16 samples around the tree is 42.4 Bq/kg and the standard deviation is 25.9 Bq/kg. This finding indicates the importance in the selection of a sampling site and the number of samples from the surface soils especially around a tree. For the vertical distribution of 137Cs activity, it was found that the concentration in the surface soil is highest, 149 Bq/kg in the hinoki stand and 101 Bq/kg in the red pine stand, and decreases with depth. The relationship between 137Cs activity and carbon content in the forest soil was investigated in two undisturbed forest stands. The relations were more precisely expressed using an exponential equation than by a linear equation. From the same forest, similar regression equations were obtained. This indicates that the distribution of 137Cs could be characterized by the organic carbon content in an undisturbed forest. It is also suggested that the coefficient values in the regression equation help to define the movement of 137Cs accompanying the decomposition of organic matter. PMID- 9851071 TI - [Congenital antithrombin III deficiency]. PMID- 9851072 TI - [Congenital hypoplasminogenemia and dysplasminogenemia (type I and type II congenital plasminogen deficiency)]. PMID- 9851073 TI - [Congenital protein C deficiency and abnormality]. PMID- 9851074 TI - [Inherited protein S deficiency]. PMID- 9851075 TI - [Activated protein C resistance]. PMID- 9851076 TI - [Plasminogen activator inhibitor-1 in thrombotic disease]. PMID- 9851077 TI - [Hageman trait]. PMID- 9851078 TI - [Heparin cofactor II deficiency]. PMID- 9851079 TI - [Antiphospholipid syndrome]. PMID- 9851080 TI - [Thrombotic thrombocytopenic purpura]. PMID- 9851081 TI - [Coagulant factor VII hyperactivity]. PMID- 9851082 TI - [Hyperfibrinogenemia]. PMID- 9851083 TI - [M0, minimally differentiated acute myeloid leukemia]. PMID- 9851084 TI - [M1, acute myeloid leukemia without maturation]. PMID- 9851085 TI - [M2, acute myeloid leukemia with maturation]. PMID- 9851086 TI - [M3, promyelocytic leukemia]. PMID- 9851087 TI - [M4, acute myelomonocytic leukemia]. PMID- 9851088 TI - [M5, acute monocytic leukemia]. PMID- 9851089 TI - [M6, erythroleukemia]. PMID- 9851090 TI - [M7, acute leukemia of megakaryocyte lineage]. PMID- 9851091 TI - [B lineage acute lymphoblastic leukemia]. PMID- 9851092 TI - [T lineage acute lymphoblastic leukemia]. PMID- 9851093 TI - [Chronic lymphocytic leukemia (CLL)]. PMID- 9851094 TI - [B-cell prolymphocytic leukemia]. PMID- 9851095 TI - [T-cell prolymphocytic leukemia]. PMID- 9851096 TI - [Chronic T-lymphocytic leukemia]. PMID- 9851097 TI - [Large granular lymphocyte leukemia]. PMID- 9851098 TI - [Adult T-cell leukemia/lymphoma]. PMID- 9851099 TI - [Adult T-cell leukemia not associated with human T-lymphotropic virus type I]. PMID- 9851100 TI - [HTLV-I-associated myelopathy]. PMID- 9851101 TI - [Hairy cell leukemia]. PMID- 9851102 TI - [Central nervous system leukemia]. PMID- 9851103 TI - [Aleukemic leukemia]. PMID- 9851105 TI - [Natural killer cell leukemia]. PMID- 9851104 TI - [Mixed lineage leukemia/hybrid leukemia/biphenotypic leukemia]. PMID- 9851106 TI - [Hypocellular acute leukemia]. PMID- 9851107 TI - [Eosinophilic leukemia]. PMID- 9851108 TI - [Basophilic leukemia]. PMID- 9851109 TI - [Plasma cell leukemia]. PMID- 9851110 TI - [Smoldering leukemia]. PMID- 9851111 TI - [Acute myeloid leukemia expressing lymphoid associated antigens]. PMID- 9851112 TI - [Myeloid antigen positive B-ALL]. PMID- 9851113 TI - [Myeloid antigen positive T-ALL]. PMID- 9851114 TI - [Secondary leukemia]. PMID- 9851115 TI - [Myeloblastoma (chloroma)]. PMID- 9851116 TI - [Refractory anemia: the entity and recent findings]. PMID- 9851117 TI - [*5q--syndrome]. PMID- 9851118 TI - [Sideroblastic anemia]. PMID- 9851119 TI - [Refractory anemia with excess of blasts (RAEB)]. PMID- 9851120 TI - [Refractory anemia with excess of blasts in transformation (RAEB-t)]. PMID- 9851121 TI - [Chronic myelomonocytic leukemia]. PMID- 9851122 TI - [Therapy-related myelodysplastic syndrome (t-MDS)]. PMID- 9851123 TI - [Trilineage myelodysplasia]. PMID- 9851124 TI - [Immunological abnormalities in myelodysplastic syndromes]. PMID- 9851125 TI - [Hodgkin's disease]. PMID- 9851126 TI - [Diffuse small lymphocytic lymphoma]. PMID- 9851127 TI - [Follicular lymphoma, medium-sized cell type and mixed type]. PMID- 9851128 TI - [Follicular lymphoma, large cell type]. PMID- 9851129 TI - [Diffuse lymphoma, medium sized cell type]. PMID- 9851130 TI - [Diffuse lymphoma, mixed type]. PMID- 9851131 TI - [Diffuse lymphoma, large cell type]. PMID- 9851132 TI - [Immunoblastic lymphoma]. PMID- 9851133 TI - [Lymphoblastic lymphoma]. PMID- 9851134 TI - [Small noncleaved cell lymphoma, Burkitt's type]. PMID- 9851135 TI - [Small noncleaved cell lymphoma, non-Burkitt's type]. PMID- 9851136 TI - [Composite lymphoma, discordant lymphoma]. PMID- 9851139 TI - [Lennert's lymphoma]. PMID- 9851137 TI - [Mycosis fungoides]. PMID- 9851138 TI - [Sezary syndrome]. PMID- 9851140 TI - [T-zone lymphoma]. PMID- 9851141 TI - [MALT lymphoma]. PMID- 9851142 TI - [Signet ring cell lymphoma]. PMID- 9851143 TI - [Intravascular lymphoma (IVL)]. PMID- 9851144 TI - [Anaplastic large cell lymphoma]. PMID- 9851145 TI - [HIV-associated lymphoma]. PMID- 9851146 TI - [Mantle cell lymphoma]. PMID- 9851147 TI - [NK-cell lymphoma]. PMID- 9851148 TI - [Null-cell lymphoma]. PMID- 9851149 TI - [Peripheral gamma delta T-cell lymphoma]. PMID- 9851150 TI - [Sarcoidosis-lymphoma syndrome]. PMID- 9851151 TI - [Necrotizing lymphadenitis]. PMID- 9851152 TI - [Multicentric Castleman's disease]. PMID- 9851153 TI - [Rosai-Dorfman disease]. PMID- 9851154 TI - [Toxoplasmic lymphadenitis]. PMID- 9851155 TI - [Cutaneous pseudolymphoma]. PMID- 9851156 TI - [Multiple lymphomatous polyposis]. PMID- 9851157 TI - [Lymphomatoid granulomatosis]. PMID- 9851158 TI - [Familial hemophagocytic lymphohistiocytosis]. PMID- 9851159 TI - [Acute lupus hemophagocytic syndrome]. PMID- 9851160 TI - [Malignancy-associated hemophagocytic syndrome]. PMID- 9851161 TI - [Autoimmune-associated hemophagocytic syndrome]. PMID- 9851162 TI - [Bacteria-associated hemophagocytic syndrome]. PMID- 9851163 TI - [Virus-associated hemophagocytic syndrome]. PMID- 9851164 TI - [Malignant histiocytosis (histiocytic medullary reticulosis)]. PMID- 9851165 TI - [Eosinophilic granuloma]. PMID- 9851166 TI - [Hand-Schuller-Christian syndrome]. PMID- 9851167 TI - [Letterer-Siwe disease]. PMID- 9851168 TI - [Gaucher's disease]. PMID- 9851169 TI - [Niemann-Pick disease]. PMID- 9851170 TI - [Tay-Sachs disease]. PMID- 9851171 TI - [Wolman disease, Fabry disease and beta-galactosidase deficiency]. PMID- 9851172 TI - [Histiocytic cytophagic panniculitis, Weber-Christian disease and nodular fat necrosis]. PMID- 9851173 TI - [IgG, IgA, Bence Jones protein (BJP) myeloma]. PMID- 9851174 TI - [IgD myeloma]. PMID- 9851175 TI - [IgM myeloma]. PMID- 9851176 TI - [IgE myeloma]. PMID- 9851177 TI - [Solitary plasmacytoma of bone]. PMID- 9851178 TI - [Extramedullary plasmacytoma]. PMID- 9851179 TI - [Smoldering multiple myeloma]. PMID- 9851180 TI - [Aggressive multiple myeloma]. PMID- 9851181 TI - [Non-secretory multiple myeloma]. PMID- 9851182 TI - [Multiple myeloma with localized tumor]. PMID- 9851184 TI - [Neuron specific enolase producing multiple myeloma]. PMID- 9851183 TI - [Amylase-producing myeloma]. PMID- 9851185 TI - [Biclonal myeloma]. PMID- 9851186 TI - [Multiple myeloma with hyperammonemia]. PMID- 9851187 TI - [Primary macroglobulinemia]. PMID- 9851189 TI - [Solitary primary macroglobulinemia]. PMID- 9851188 TI - [Leukemic macroglobulinemia]. PMID- 9851190 TI - [X-linked hyper IgM syndrome]. PMID- 9851191 TI - [Heavy chain disease]. PMID- 9851192 TI - [Monoclonal gammopathy of undetermined significance (MGUS)]. PMID- 9851193 TI - [Plasma cell dyscrasia associated with polyneuropathy and endocrinological abnormality (Takatsuki disease)]. PMID- 9851194 TI - [Myeloma kidney]. PMID- 9851195 TI - [Amyloidosis associated with multiple myeloma]. PMID- 9851196 TI - [Primary amyloidosis]. PMID- 9851197 TI - [Secondary (AA) amyloidosis]. PMID- 9851198 TI - [Familial amyloidotic polyneuropathy]. PMID- 9851199 TI - [Familial Mediterranean fever]. PMID- 9851200 TI - [Solitary nodular pulmonary amyloidosis]. PMID- 9851201 TI - [Cardiac amyloidosis]. PMID- 9851202 TI - [Cerebral amyloidosis]. PMID- 9851203 TI - [Cutaneous amyloidosis]. PMID- 9851204 TI - [Dialysis-related amyloidosis]. PMID- 9851205 TI - [Single component monoclonal cryoglobulinemia (type I)]. PMID- 9851206 TI - [Monoclonal-polyclonal mixed cryoglobulinemia (type II)]. PMID- 9851207 TI - [Polyclonal mixed cryoglobulinemia (type III)]. PMID- 9851208 TI - [Essential hypergammaglobulinemia]. PMID- 9851209 TI - [Hypergammaglobulinemic purpura]. PMID- 9851210 TI - [Polyclonal hypergammaglobulinemia]. PMID- 9851211 TI - [Benign IgM gammopathy]. PMID- 9851212 TI - [Biclonal gammopathy]. PMID- 9851213 TI - [IgG3 deficiency]. PMID- 9851214 TI - [Hyper-beta 2-microglobulinemia]. PMID- 9851215 TI - [Congenital abetalipoproteinemia (Bassen-Kornzweig syndrome)]. PMID- 9851216 TI - [Tangier disease (familial high density lipoprotein deficiency)]. PMID- 9851217 TI - [Congenital analbuminemia]. PMID- 9851218 TI - [Alloalbuminemia]. PMID- 9851219 TI - [Acquired hypoalbuminemia]. PMID- 9851220 TI - [The syndrome of the sea-blue histiocyte]. PMID- 9851221 TI - [Sweet's syndrome]. PMID- 9851222 TI - [Systemic capillary leak syndrome]. PMID- 9851223 TI - [Tn syndrome]. PMID- 9851224 TI - [Light chain deposition disease]. PMID- 9851225 TI - [Acquired thrombophilia]. PMID- 9851226 TI - [Fetomaternal and twin to twin (feto-fetal) transfusion syndrome]. PMID- 9851227 TI - [Engraftment syndrome in hematopoietic stem cell transplantation]. PMID- 9851228 TI - [Chronic GVHD]. PMID- 9851229 TI - [Transfusion associated graft versus host disease (TA-GVHD)]. PMID- 9851230 TI - [Hyperviscosity syndrome]. PMID- 9851231 TI - [Thrombotic microangiopathy in hematopoietic stem cell transplantation]. PMID- 9851232 TI - Neurological examination of the newborn. PMID- 9851233 TI - Cognitive and educational outcome of very-low-birthweight children in early adolescence. AB - A cohort of 138 very-low-birthweight (VLBW) 12-year-old children and matched control children were assessed on objective cognitive and educational measures. School performance was rated by teachers and by the children themselves. VLBW children were shown to have lower IQ scores, and poorer scores on all objective educational measures compared with control children. Controlling for IQ differences, mathematics and reading-comprehension scores remained significantly lower for VLBW children. Teachers rated VLBW children lower in all curriculum areas. Significantly more VLBW children were found to be 'failing' in one or more subject and an increased proportion compared with the control children had received remedial education. The VLBW group showed no evidence of 'catch up' between 6 and 12 years of age. Multiple regression analyses were used to identify predictors of cognitive and educational outcome. The duration of mechanical ventilation in the neonatal period was inversely related to outcome. Full-Scale IQ at 6 years, motor-skills score at 6 years, and head circumference at 12 years all predicted outcome at 12 years, as did maternal education, family income and size. Individually, many VLBW children perform satisfactorily, but as a group VLBW children appear to be at a long-term disadvantage to peers in the areas of cognitive and educational performance. PMID- 9851234 TI - Neuromotor ability in 5- to 7-year-old children with very low or extremely low birthweight. AB - This study was intended to determine the effects of extremely low birthweight (ELBW, 500 to 999 g) and very low birthweight (VLBW, 1000 to 1499 g) on neuromotor ability in 5- to 7-year-old children. Fourteen ELBW and 20 VLBW children were compared with 24 term control children of normal birthweight (NBW, >2500 g). Using quantitative assessment instruments, the following data were collected: maximal cycling speed during 30 seconds of cycling at 'zero' resistance, simple reaction time of the legs, and performance on components of a whole-body coordination test. The main findings were a slower reaction time, lower maximal cycling speed, and lower coordination scores in the ELBW group compared with the NBW group and, for some variables, with the VLBW group. The reduced motor performance in these children appears for the most part to be a reflection of impaired neuromotor control and motor development, rather than merely a smaller body or muscle size. PMID- 9851235 TI - Is the long-term outcome of children following febrile convulsions favorable? AB - The study comprised 80 children aged 6 to 9 years with a history of febrile convulsions. A neurological examination, an interview to assess psychiatric anomalies, and a series of neuropsychological tests were performed on patients with previous febrile convulsions and on matched healthy controls. Children with non-febrile seizures or CNS infections were excluded. Recurrence of febrile seizures in the study group was 41% (N=33), 18 children (22%) had prolonged febrile convulsions, six (7.5%) patients and two controls showed discrete neurological abnormalities. Behavioral anomalies were exhibited by 22% of the patients and 6% of the healthy children. The neuropsychological test results did not demonstrate significant differences between the children with febrile convulsions and the healthy controls. However, in children with prolonged febrile convulsions, non-verbal intelligence was found to be significantly lower as compared with children with simple febrile seizures and with controls. None of the other parameters tested yielded any differences between patients and controls. Children with multiple recurrences of febrile convulsions performed poorer in all tests when compared with children with only one febrile seizure or with controls. Other factors such as a positive family history of epilepsy, age at onset of febrile convulsions, or duration of the seizure were not found to be of prognostic significance. PMID- 9851236 TI - Use of proprioception in normal and clumsy children. AB - This study investigates the relation between performance on simple tasks dependent on proprioception, and performance of complex perceptual-motor skills in clumsy children and age-matched control children. One hundred and forty-six right-handed children aged between 5 and 8 years were tested on non-visual aiming, non-visual posture matching, the Kinaesthetic Sensitivity Test (KST), and the Movement Assessment Battery for Children (ABC). Half of the children had scores below the 15th centile on the Movement ABC and were classed into the developmental coordination disorder (clumsiness) group. Scores on the proprioceptive tasks were used to predict performance on complex tasks of the subscales of the Movement ABC (manual dexterity, ball skills, and balance). Specific relations were found between the proprioceptive tasks and the subscales of the Movement ABC, but the KST did not predict differences in motor skills, and no relation was found between tasks carried out without vision. Simple non-visual movement tasks do predict performance in more complex skilled tasks but this is affected by many task features rather than simply the reliance on proprioception for information about movement. PMID- 9851237 TI - Eye-movement disorders and visual-perceptual impairment in diplegic children born preterm: a clinical evaluation. AB - The visual-motor behaviour of 15 preterm diplegic children and 50 control children (age range 4 to 7 years) was recorded on video as they performed a visual-perceptual task (an adaptation of the Animal House subtest of the Wechsler Preschool Primary Scale of Intelligence). The following parameters were analysed and scored: time to perform task; omissions; figure-colour association; sequence direction; sequential scanning order; accuracy of fitting target; and number of anticipatory saccadic movements to next target. The ability of the control children to perform the task improved significantly with age, as measured by performance time, mistakes in sequence direction and scanning order, accuracy of target fitting, and number of anticipatory saccadic movements. The scores of children with diplegia were not related to age and were poorer overall than those of the control group. Children with diplegia made significantly more mistakes of sequence direction and scanning order, and significantly fewer anticipatory saccadic movements than the control group. These results indicate that visual perceptual impairment in diplegic children born preterm is not attributable only to sensory visual loss and to fine manipulation difficulties but is also related to difficulties in eye movements and in using anticipatory control to process information. PMID- 9851238 TI - Linear growth velocity in children with cerebral palsy. AB - Retrospective analysis of an anthropometric database collected prospectively in children with cerebral palsy (CP) was carried out to evaluate linear growth velocity and identify risk factors for poor linear growth. Growth velocity measures were compared with published norms for prepubertal growth velocity and z scores were calculated. Mean growth velocity z score (Gvz) was -0.97+/-1.9. Boys grew more slowly than girls (mean Gvz = -1.5+/-1.9 versus -0.3+/-1.7 P = 0.003). Gvz did not correlate with type of CP or presence/absence of microcephaly. Young age was a risk factor for poor linear growth (mean Gvz = -2.40+/-2.6 for children under 2 years of age versus -0.76+/-1.9 ages 2 to 6 and -0.77+/-1.6 ages 6 to 10, P = 0.04). Children at nutritional risk (triceps skinfold thickness < or =55%) grew poorly (mean Gvz = -1.46+/-1.5 versus -0.30+/-1.8, P = 0.01). For children over 2 years, those with cognitive impairments grew more slowly than those with normal cognition (mean Gvz = -1.25+/-1.9 versus -0.12+/-1.8, P = 0.02) and non ambulatory children grew more slowly than ambulatory children (mean Gvz = -1.20+/ 1.5 versus -0.35+/-1.9, P = 0.03). Prepubertal children with CP grow more slowly than expected compared with age- and sex-based standards. Sex, age, cognitive impairment, ambulatory status, and nutritional state are factors which may contribute to slow growth. These results add to the growing data that children with CP have unique growth patterns. Further study is needed to clarify the factors which contribute to poor linear growth in this population. PMID- 9851239 TI - Evaluating motor function in children with Down syndrome: validity of the GMFM. AB - The Gross Motor Function Measure (GMFM) was developed and validated originally by Russell and colleagues as an evaluative assessment of gross motor function in children with cerebral palsy (CP). The present study reports the results of reliability and validity testing of the GMFM for use with children with Down syndrome (DS). One hundred and twenty-three children with DS were assessed twice over a 6-month period, using the GMFM and the motor scale of the Bayley Scales of Infant Development - second edition (BSID-II). In addition to the usual method of scoring the GMFM using only observed motor behaviours (standard score), parent reports of children's activities not seen by the assessor on the day of testing were also obtained and a second score (reported score) was calculated for each GMFM assessment. Test-retest and interrater reliabilities were excellent (all >0.90). Observed correlations between change on GMFM and judgements of change made independently by parents, intervenors, and masked video raters were lower than hypothesized. However, the pattern of change scores in predefined age and severity subgroups supported the contention that the GMFM was able to detect differential amounts of change as predicted. The GMFM was shown to be relatively more responsive to change in gross motor function than the motor scale of the BSID-II. The 'reported' scores on the GMFM demonstrated better evidence of reliability, validity, and responsiveness than the standard scoring method and this approach is recommended for use when assessing children with DS. PMID- 9851240 TI - Two males with childhood disintegrative disorder: a prospective 14-year outcome study. AB - A prospective 14-year outcome study of two children meeting DSM-IV criteria for childhood disintegrative disorder is presented. Their ages at first evaluation were 4 years 7 months and 6 years 3 months. Both are now adults and continue to have a severe pervasive developmental disorder, mental retardation, seizure disorder, and are non-verbal. Both require residential care. PMID- 9851241 TI - Neurologic evaluation of the newborn infant: definition and psychometric properties. PMID- 9851242 TI - Oral angle elevation induced by tapping stimulation of the supraorbital ridges: a synkinesis-like movement implies severe cerebral and brainstem dysfunction. PMID- 9851243 TI - Selective dorsal rhizotomy studies. PMID- 9851244 TI - 5E10: a prostate-specific surface-reactive monoclonal antibody. AB - We created mAb that reacted solely with prostate epithelial cells. The strategy involved immunizing mice with a mixture of six different prostatic carcinoma cell lines and selecting by flow cytometry only those antibodies that bind whole cells. The primary screening was performed using a mixture of all six prostate cell lines used in immunization together with six non-prostate cell lines in the same tube. Antibodies that gave a bimodal pattern of surface staining were selected for further evaluation. The most attractive clone, designated 5E10, produced IgG1 mAb that recognized four of the six prostatic cell lines and did not react with non-prostate tumor cell lines, peripheral blood and bone marrow mononuclear cells and endothelial and bone marrow stromal cells. 5E10 mAb reacted with both benign and malignant prostate in eight of eight histological samples and no reactivity was noted with non-prostate normal tissues. The 5E10 antigen is a transmembrane glycoprotein with a molecular weight of 110 kDa and the epitope recognized by 5E10 is extracellular. Ongoing studies are exploring the nature of this antigen in more depth. PMID- 9851245 TI - Inhibition of tumor angiogenesis by roxithromycin, a 14-membered ring macrolide antibiotic. AB - We examined the effects of roxithromycin, a 14-membered ring macrolide antibiotic, on tumor angiogenesis using a mouse dorsal air sac model. The inhibitory effect of roxithromycin was dose-dependent and 100 mg/kg of roxithromycin administered intraperitoneally twice a day reduced the dense capillary network area to about 20% of the control. However, at concentrations of up to 50 microM, roxithromycin had no effect on lung cancer cells and human vascular endothelial cell growth and lung cancer cell production of the angiogenesis-inducing factors interleukin-8 and vascular endothelial growth factor. Roxithromycin at concentrations greater than 20 microM inhibited endothelial cell migration and tube formation. PMID- 9851246 TI - Proliferating cell nuclear antigen (PCNA) immunolabeling as a prognostic factor in invasive ductal carcinoma of the breast in Taiwan. AB - To evaluate the prognostic significance of proliferating cell nuclear antigen (PCNA) in breast cancer, an immunohistochemical assay was performed in 150 patients with invasive ductal carcinomas. The PCNA labeling index (PCNA-LI) was classified into two subgroups at a cut-off point of 45% that gave the best prognostic estimates for PCNA in survival analyses. Seventy-eight tumors had a low PCNA-LI of < or =45% and 72 tumors had a high PCNA-LI of >45%. A high PCNA-LI correlated significantly with p53 overexpression (P<0.03), positive axillary node (P<0.04), short disease-free survival (P<0.001) and overall survival (P<0.0002), but not with other factors. In multivariate analysis, the PCNA-LI predicted the disease-free (P<0.008) and overall survival (P = 0.0007) independently. Our study indicates that the PCNA-LI has independent prognostic value. PMID- 9851247 TI - Inhibition of polyposis in the small intestine of BALB/c mice by intestinal bacteria. AB - The incidence of polyposis and the number of polyps per mouse were significantly lower in conventionalized (CVz) mice than in germ-free (GF) mice. There was no significant difference in the average number of polyps between GF and gnotobiotic (GB) mice monoassociated with the various strains of intestinal bacteria. However, the incidence of polyposis and the number of polyps per mouse were significantly lower for mice associated with either chloroform-resistant bacteria (CRB) or fusiform bacteria (FB) than for GF mice. This study demonstrated that polyposis was suppressed by FB and CRB in the small intestine of BALB/c mice. PMID- 9851248 TI - Angiogenesis induced by urothelial cells (HCV-29) and their v-ras and v-raf transfectants. AB - The angiogenic ability of human urothelial cells (HCV-29) and their v-ras and v raf transfectants was studied. The most pronounced angiogenesis, observed in vivo, induced v-raf-transfected cells. The lowest degree of induction of neovascularization presented cells of the parental line. The increased extent of angiogenesis correlated with the presence of VEGF mRNA as measured by RT-PCR as well as the level of VEGF as visualized by the method of Western blotting. PMID- 9851249 TI - Does the amount of an antitumor agent entrapped in liposomes influence its tissue distribution and cell uptake? AB - The effects of the amount of a drug entrapped in liposomes and polyethyleneglycol (PEG) modification on the tissue distribution in vivo and cell uptake in vitro have been examined. An increase in the amount of doxorubicin (DOX) entrapped in liposomes induced an increase in the DOX level in the plasma and tumor and a decrease in this level in the liver. The high amount of DOX entrapped demonstrated the usefulness of DOX liposomes for tumor cell uptake in vitro. The cell uptake of the liposomes depended on additional amounts of DOX and liposomal lipid. Furthermore, PEG modification of the surface of the liposomes facilitated the initial rate of liposome uptake into the tumor cells. This facilitation was attributed to the lipohydrophilic property of PEG and the fixed aqueous layer around the liposomes. Therefore, a higher amount of DOX entrapped in liposomes and PEG modification have been confirmed to be beneficial. PMID- 9851250 TI - Different ras gene mutational frequencies in thyroid papillary carcinomas in Japan and Thailand. AB - The incidence and pattern of ras oncogene mutations in human malignancies demonstrate geographic and racial differences. For example, specificity of alterations is found in cholangiocellular carcinomas in Thai patients with a different etiology from those in Japanese patients. In the present study, a comparison of ras gene mutations in thyroid papillary carcinomas from Japanese and Thai patients was performed using single-strand conformation polymorphism and direct sequencing analyses. The incidence of ras mutation differed markedly in Japanese (two of 24 carcinomas, 8.3%) and Thai (five of 10 carcinomas, 50%) patients. In addition, all but one ras mutation occurred at codon 12 of the K-ras gene in the Thai cases. These results suggest that thyroid cancers in Thailand may be due to specific genetic and/or environmental factors. PMID- 9851251 TI - Overexpression of cyclin D1 and cdk4 in tumorigenesis of sporadic hepatoblastomas. AB - Abnormality of the cyclin D1/cdk4/p16INK4a/pRb pathway during tumorigenesis has recently been reported. Hepatoblastoma is a rare malignant liver tumor of childhood, but underlying abnormalities of cell-cycle regulating protein remain to be elucidated. The expression of cyclin D1, cdk4, p16 and retinoblastoma gene product (pRb) was studied by immunohistochemistry in 17 paraffin-embedded tissues consisting of both tumor and corresponding non-neoplastic tissues. Tumor tissues showed overexpression of cyclin D1 (13/17, 76%) and cdk4 (15/17, 88%). Eleven cases showed co-overexpression of both cyclin D1 and cdk4. No abnormal p16 or pRb expression was noted. In the group with a high score (+4) for cyclin D1 expression, a positive correlation with tumor recurrence was noted (P = 0.043). These data suggest that overexpressed cyclin D1 and cdk4 protein might play an important role in the tumorigenesis of hepatoblastoma and that in the group with high cyclin D1 expression, tumor recurrence may be more frequent. PMID- 9851252 TI - Increased telomerase activity in intrahepatic cholangiocellular carcinomas induced by N-nitrosobis(2-oxopropyl)amine in hamsters. AB - Telomerase activities in intrahepatic cholangiocarcinomas induced by N nitrosobis(2-oxopropyl)amine (BOP) in female hamsters were determined using a telomeric repeat amplification protocol (TRAP) assay followed by densitometric quantification. Each determination was repeated to confirm the results and telomerase activity was also detected by gel electrophoresis. An increase was evident in all of 10 cholangiocarcinomas examined, with levels ranging from 2.48 to 4.40 times the normal liver value by densitometric quantification. This finding of a consistent increase suggests that telomerase activation is involved in the development of intrahepatic cholangiocarcinomas and immortalization of cancer cells. PMID- 9851253 TI - Inhibition of K-ras-transformed rodent and human cancer cell growth via induction of apoptosis by irreversible inhibitors of Ras endoprotease. AB - Proteolytic removal of the carboxyl terminal tripeptide of Ras oncoproteins is important in the Ras function. Two chloromethyl ketones, BFCCMK and UM96001, designed to be the Ras C-terminal sequence-specific endoprotease inhibitors, at low micromolar concentrations (5.0 microM), potently inhibit the growth of ras transformed rodent and human cancer cells, whereas untransformed NIH/3T3 cells are not affected under the same conditions. Furthermore, BFCCMK and UM96001 block more than 98% of the anchorage-independent clonogenic growth of ras-transformed rat and human cancer cells at low micromolar concentrations. The blocking of cancer cell growth may be due to the selective induction of apoptosis of ras transformed cells by these inhibitors. These results provide the first experimental evidence that the endoproteolysis of Ras oncoproteins is important for the growth and apoptosis of ras-transformed cancer cells. Therefore, the Ras C-terminal sequence-specific endoprotease may be a potential new target for the treatment of human cancers induced by ras mutations. PMID- 9851254 TI - Transcriptional activity of quinone methides derived from the tumor promoter butylated hydroxytoluene in HepG2 cells. AB - Butylated hydroxytoluene (BHT) is a pulmonary toxin and tumor promoter in mice presumably due to the formation of two quinone methides (QMs) that alkylate cellular nucleophiles. The activation of stress genes by these electrophilic metabolites was investigated with an assay system consisting of 14 recombinant cell lines derived from the human hepatoma line HepG2, each carrying a unique promoter or response element construct fused to the reporter gene for chloramphenicol acetyl transferase (CAT). The largest responses to QMs occurred in cells containing either the metallothionein IIA, glutathione S-transferase Ya, or 70 kDa heat shock protein promoter, or the xenobiotic response element. The other cell lines exhibited only small or no effects. These results are consistent with transcriptional activities reported for several other electrophiles known to undergo covalent interactions with proteins. PMID- 9851255 TI - Antitumour activity of diallyl sulfide on polycyclic aromatic hydrocarbon-induced mouse skin carcinogenesis. AB - Diallyl sulfide (DAS), a major flavour component of garlic, is known to modulate xenobiotic metabolism and possess antitoxic, bactericidal, antineoplastic, hypolipidemic and hypocholesteromic effects. In the present study, the anticarcinogenic activity of DAS on a 7,12-dimethylbenzanthracene (DMBA)- or benzo[a]pyrene (B(a)P)-induced mouse skin model of carcinogenesis was evaluated. DAS was applied topically either 1 h prior to or 1 h after the administration of DMBA or B(a)P. A significant protection from neoplasia was observed in DAS- and DMBA/B(a)P-exposed animals when DAS was applied topically compared to the animals exposed only to DMBA/B(a)P. In the animals where DAS was applied 1 h prior to the application of DMBA, a lower magnitude of neoplasia was recorded in terms of the cumulative number of tumours and average number of tumours per mouse during the entire period of study (28 weeks) compared to the animals exposed to DAS 1 h later, while in B(a)P-exposed animals, the antitumorigenic potential of DAS was more evident in the mice treated with DAS 1 h after the B(a)P exposure compared to the animals treated with DAS 1 h prior to B(a)P. The antitumour activity of DAS was of a much higher magnitude in B(a)P-induced carcinogenesis in comparison to animals exposed to DMBA in terms of tumour incidence, cumulative number of tumours and average number of tumours per mouse. The results suggest that DAS has a protective effect in PAH-induced mouse skin carcinogenesis. PMID- 9851256 TI - p53 gene mutations in rectal cancer associated with schistosomiasis japonica in Chinese patients. AB - Mutations in p53 tumor suppressor gene were examined in 44 Chinese patients with rectal cancer, including 22 cases with advanced schistosomiasis japonica and 22 cases without schistosomiasis. In schistosomal rectal cancer (SRC), 13 mutations were found in 10 cases, which included 11 base-pair substitutions and two deletions. Of 11 base substitutions, nine were transitions and two were transversions and seven of them were located at CpG dinucleotides. In non schistosomal rectal cancer (NSRC), 13 mutations were found in nine cases, all of which were base-pair substitutions. Of 13 substitutions, 10 were transitions and three were transversions and three of them were located at CpG dinucleotides. The proportion of base-pair substitutions at CpG dinucleotides was higher in SRC patients than in NSRC patients, although this was not statistically significant (P = 0.054). Point mutation was frequent at codon 248 in SRC. A higher frequency of arginine missense mutations was observed in SRC than in NSRC. These observations suggest that the mutations in SRC are the result of genotoxic agents produced endogenously through the course of schistosomiasis japonica. PMID- 9851257 TI - High alkaline phosphatase activity in 3p-induced human renal cancer cells. AB - Recent studies have suggested that a tumor suppressor gene is located on the short arm of chromosome 3 (3p) for tumorigenesis of sporadic renal cell carcinoma. In this study, we introduced 3p from normal human fibroblasts into the renal cancer cell line YCR and this introduction clearly resulted in inhibited growth in vitro and in vivo. Furthermore, we investigated several enzyme activities of original YCR cells and 3p-induced clones (YCR-151 and YCR-152) and found that only alkaline phosphatase activity in 3p-induced clones was significantly increased compared to that of the original YCR cells. PMID- 9851258 TI - Glucocorticosteroids in rheumatoid arthritis: lessons for the future. PMID- 9851259 TI - From criticism to creativity: the genesis of the ARC/BSR Clinical Trials Group. PMID- 9851260 TI - Skin hyperreactivity of Behcet's patients (pathergy reaction) is also positive in interferon alpha-treated chronic myeloid leukaemia patients, indicating similarly altered neutrophil functions in both disorders. AB - Typical manifestations of Behcet's disease (BD) and a positive pathergy reaction were observed in a few patients with chronic myeloid leukaemia (CML) on interferon alpha (IFN-alpha) therapy and the significance of this observation was assessed in a prospective study. The skin pathergy test was applied to 15 patients with CML prior to IFN-alpha therapy, 29 patients with CML following IFN alpha therapy and 30 patients with BD. Twenty-five patients with inflammatory arthropathies (IA), 20 patients with recurrent oral ulcers (ROU), 23 patients treated with IFN-alpha for various disorders and 20 normal individuals were also studied as control groups. The pathergy reaction was positive in nearly a quarter of IFN-alpha-treated CML cases (24%) as well as one-half of the patients with BD (50%). All CML patients prior to IFN-alpha treatment and all patients using IFN alpha for other diseases were negative for the pathergy reaction. These results may indicate a similarly altered neutrophil function in both BD and IFN-alpha treated CML patients. PMID- 9851261 TI - Evaluation of humeral head erosions in rheumatoid arthritis: a comparison of ultrasonography, magnetic resonance imaging, computed tomography and plain radiography. AB - The value of ultrasonography (US), magnetic resonance imaging (MRI), computed tomography (CT) and plain radiography (PR) in detecting bone erosions on the humeral head was evaluated in a study of 26 in-patients (26 shoulders) with rheumatoid arthritis (RA). MRI depicted humeral erosions in 25 (96%), US in 24 (92%), CT in 20 (77%) and PR in 19 (73%) of the 26 shoulders. MRI and US were superior to CT in detecting small erosions. US was the most sensitive method to show surface erosions on the greater tuberosity. US, CT and MRI detected large erosions quite similarly. PR frequently missed small erosions. In the evaluation of early erosions in the rheumatoid shoulder, US and MRI are more sensitive methods than the traditionally used PR. US and MRI are suitable for the evaluation of soft-tissue involvement in the rheumatoid shoulder, but also for the detection of bone erosions of the humeral head. PMID- 9851262 TI - Early effective suppression of inflammation in rheumatoid arthritis reduces radiographic progression. AB - OBJECTIVE: To evaluate the effect of early 'aggressive' drug treatment on radiographic progression in patients with recent-onset rheumatoid arthritis (RA), compared to conventional stepwise increasing intensity of treatment. DESIGN: Prospective follow-up study with an experimental group and a historical control group both divided into a high-risk subgroup and a low-risk subgroup, based on prognostic factors. The effect of the 'aggressive' and the conventional treatment strategy was compared between both high-risk groups; the low-risk groups, both treated according to the conventional treatment strategy, were used to ensure internal consistency between the experimental and the historical groups. PATIENTS: A total of 228 consecutive patients with recent-onset RA (complaints < 1 yr at study entry). METHODS: The 'aggressive' drug treatment consisted of institution of relatively fast-acting disease-modifying anti-rheumatic drugs (DMARDs) (sulphasalazine, methotrexate) immediately after diagnosis, and rapid adjustment of dosage and/or drug in the case of insufficient response as measured by a change in C-reactive protein (CRP) level. Radiographic damage was assessed according to a modified version of Sharp's method and cumulative disease activity expressed as CRP-area under the curve (CRP-AUC). The occurrence of side-effects was also evaluated. RESULTS: After 2 yr of follow-up, comparison of the two high risk subgroups showed the radiographic progression in the 'aggressively' treated subgroup to be significantly lower than that in the control group [Sharp score: median (range) 26 (0-100) vs 35 (1-188); P = 0.03]. Cumulative CRP values were also significantly lower than in the control high-risk subgroup [CRP-AUC: median (range) 1963 (212-8515) vs 3025 (46-15 632) mg.week/1; P = 0.002). This was achieved without an increase in the occurrence of side-effects. There was no difference between the two low-risk subgroups with regard to entry characteristics, CRP-AUC values or radiological progression, indicating comparability between the two groups. CONCLUSION: Early 'aggressive' drug treatment, using sulphasalazine and/or methotrexate, aimed at reduction of the CRP level, significantly reduces the (rate of) radiographic progression in RA. PMID- 9851263 TI - Autoantibodies against human calpastatin in rheumatoid arthritis: epitope mapping and analysis of patient sera. AB - Autoantibodies against calpastatin have recently been described to be highly prevalent in sera of patients with rheumatoid arthritis (RA). When the sera of 45 patients with RA were analysed for autoantibodies against calpastatin by a newly developed enzyme-linked immunosorbent assay (ELISA), only four sera (8.9%) tested positive, which is not significantly different from the frequency observed in healthy controls. Since the ELISA is based on a synthetic peptide containing the C-terminal 27 amino acids of calpastatin bound to the solid phase, this negative result might be the consequence of the small antigen used. Therefore, a systematic analysis of the epitopes for autoantibodies in calpastatin was performed using sera from RA patients and healthy individuals. Recombinant fusion proteins containing fragments of calpastatin or the complete protein were produced and sera analysed by Western blots. In the N-terminal portion (amino acids 1-369), at least two major epitopes exist, against which 65% of normal sera as well as 76% of RA sera show reactivity in Western blot assays. These epitopes are not useful for clinical diagnostics. Only five out of 45 (11.1%) RA sera reacted against the C-terminal portion (amino acids 363-708) of calpastatin, while four out of 52 (7.7%) control sera showed reactivity. Three of the five RA sera and two out of four control sera had autoantibodies against the C-terminal 27 amino acids of calpastatin. These three patient sera had already been tested positive in the ELISA. The fourth patient positive in the ELISA was Western blot negative. The differences between the group of RA patients and controls are not statistically significant. When the clinical characteristics of the four patients with autoantibodies against the carboxyl end of calpastatin were analysed, it became apparent that all four had significantly elevated C-reactive protein (>50 mg/l). This observation might indicate that calpastatin autoantibodies are found in RA patients with more active disease. Thus, while the majority of RA patients do not have an increased prevalence of calpastatin autoantibodies, it cannot be ruled out definitively that a small subgroup may be characterized by autoantibodies to the C-terminus of calpastatin. PMID- 9851264 TI - Insulin-like growth factor-I and insulin-like growth factor binding protein-3 serum levels in ankylosing spondylitis. AB - OBJECTIVE: Low bone mass, vertebral osteopenia and fractures have been described in patients with ankylosing spondylitis (AS), but the aetiology of this osteoporosis (OP) remains unknown. Insulin-like growth factor-I (IGF-I), a bone promoting peptide, may be considered as reflecting osteoblast function as well as its main binding protein, insulin-like growth factor binding protein-3 (IGFBP-3). Both were found to be decreased in post-menopausal women and male patients with idiopathic OP. In this study, we aimed to measure the circulating IGF-I and IGFBP 3 in AS patients. METHODS: Thirty-three AS patients were compared to 23 healthy controls. Bone mineral density (dual X-ray absorptiometry) was measured at the spine and the femoral neck. We determined the serum levels of growth hormone (GH), insulin, glycaemia, and the IGF-I and IGFBP-3 serum concentrations. RESULTS: A lowered lumbar spine bone mineral density was found in the AS group (AS: 0.946 g/cm2, controls: 1.02 g/cm2; P = 0.05). AS patients had a higher glycaemia than controls, but results were in the normal range. There were no significant differences in the mean values for GH and insulin. Mean IGF-I serum levels were 218.3 ng/ml (+/-72.4) in patients and 212.1 (+/-71.1) in controls (P = 0.75). The serum concentrations of IGFBP-3 were significantly lower in AS (3.29+/-0.6 microg/ml) than in healthy subjects (3.63+/-0.6 microg/ml; P = 0.05). There was a negative correlation between the serum IGFBP-3 concentration and erythrocyte sedimentation rate (r = -0.39; P = 0.025). CONCLUSIONS: Since IGFBP-3 is an important cofactor for IGF-I and modulates its bioavailability and activity in bone, these data suggest that osteoblast cell function could be impaired in AS. Inflammation could play a role in this IGFBP-3/IGF-I axis involvement. However, further studies are warranted to determine the role of the other growth factors and their binding proteins in the OP of AS. PMID- 9851265 TI - Pogosta disease: clinical observations during an outbreak in the province of North Karelia, Finland. AB - OBJECTIVE: To characterize the clinical picture of Pogosta disease. METHOD: The data of 73 patients who had had Pogosta disease in 1981 and who then had been seen by a local physician in North Karelia were analysed. RESULTS: The main manifestations were fever (23%), rash (88%) and joint symptoms (93%). The joint symptoms in some patients lasted for several months and were severe enough to cause immobilization. The clinical picture was identical in those patients who had a definite serological diagnosis and those who did not have a detectable antibody response. CONCLUSION: The symptoms of Sindbis virus-induced Pogosta discase consist of fever, rash and joint symptoms, whic may be severe and prolonged. PMID- 9851266 TI - Improvements in quadriceps sensorimotor function and disability of patients with knee osteoarthritis following a clinically practicable exercise regime. AB - OBJECTIVE: Quadriceps sensorimotor dysfunction may be important in the pathogenesis of knee osteoarthritis (OA) and a determinant of disability. Exercise regimes can increase quadriceps strength, but whether this improves proprioception and reduces disability is uncertain. Moreover, research regimes involve protracted treatment which is clinically impracticable. METHODS: We compared quadriceps sensorimotor function and disability in 60 patients with knee OA, before and after an exercise regime, with a control group (n = 37) who did not exercise. RESULTS: Exercise improved quadriceps strength (mean change, 95% CI; 73 N, 26-19 N), voluntary activation (14%, 5-20%), knee joint position sense (0.6 degrees, 0.1-1.8 degrees), and reduced the Lequesne Index (3.5, 0.5-4) and aggregate time of four activities of daily living (8.4 s, 0.2-16.7 s). At 6 month follow-up, these improvements were maintained. The parameters of the control group were unchanged. CONCLUSIONS: These results substantiate the association between quadriceps sensorimotor dysfunction and disability, emphasizing the importance of quadriceps exercise in the management of knee OA. The regimen is relatively brief and clinically practicable, but could be adapted to make it more cost effective. PMID- 9851267 TI - Increased serum levels of soluble vascular cell adhesion molecule-1 and E selectin in patients with systemic sclerosis. AB - OBJECTIVE: To determine the serum levels of soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble E-selectin (sE-selectin) in patients with systemic sclerosis (SSc). METHOD: Serum samples from 80 patients with SSc and 20 healthy control subjects were examined by a sensitive enzyme-linked immunosorbent assay. RESULTS: The serum levels of sVCAM-1 and sE-selectin were significantly higher in the patients with SSc than in the healthy controls. The serum levels of sVCAM-1 were correlated with the presence of pulmonary fibrosis, joint involvement and elevated erythrocyte sedimentation rate levels. The serum levels of sE-selectin were correlated with the presence of pulmonary fibrosis. CONCLUSION: These results suggest that endothelial activation is involved in the development of this disease. PMID- 9851268 TI - Acetabular dysplasia and hip osteoarthritis in Britain and Japan. AB - OBJECTIVE: Geographic differences in the prevalence of hip osteoarthritis (OA) have been ascribed to differences in the frequency of acetabular dysplasia among different ethnic groups. However, there are few data on the shape of the acetabulum in various populations around the world. We examined this issue in samples of pelvic radiographs from Britain and Japan. METHODS: Measurements were made on the pelvic radiographs of 1303 men and 195 women, aged 60-75 yr, who attended for i.v. urography in two British centres. These were compared with 99 men and 99 women aged 60-79 yr who were included in a population-based study in a rural community in Japan, and who agreed to undergo standardized pelvic radiography. Acetabular dysplasia was assessed by morphometric measurement of the centre-edge (CE) angle and acetabular depth. RESULTS: The mean CE angle among men was 36 degrees (95% CI 35-37 degrees ) in Britain and 31 degrees (95% CI 29-32 degrees ) in Japan; that in women was 37 degrees (95%, CI 36-38 degrees ) in Britain and 31 degrees (95% CI 29 33 degrees ) in Japan. The mean values of acetabular depth were also significantly (P < 0.001) lower in Japan than in Britain. However, the prevalence of hip OA was lower in Japan (0% in men, 2% in women) than in Britain ( 11% in men, 4.8 / in women). In a random effects model, there were negative relationships between measures of acetabular dysplasia and minimum joint space among individuals. CONCLUSIONS: We conclude that there are marked differences in pelvic morphometry between Britain and Japan. The acetabular dimensions of Japanese subjects are considerably shallower than those of their British counterparts of similar age and sex. Nevertheless, hip OA is more frequent in Britain than in Japan. Further studies are required on the risk factors for hip OA in Oriental populations, in order that the aetiology of this disorder can be better understood. PMID- 9851269 TI - Hormone replacement therapy, other reproductive variables and symptomatic hip osteoarthritis in elderly white women: a case-control study. AB - BACKGROUND: Recent epidemiological studies suggest that post-menopausal hormone replacement therapy might reduce the risk of hip osteoarthritis (OA) in women. However, the association of the disorder with other reproductive variables is controversial. We addressed this issue in a population-based case control study among 413 female cases and 413 age- and sex-matched controls. METHODS: A total of 413 women listed for hip replacement because of primary OA over an 18 month period were compared with an equal number of controls selected from the general population and individually matched for age and general practice. Information about reproductive variables was obtained by questionnaire administered at interview. RESULTS: The risk of hip OA was significantly elevated among women who had had an oophorectomy (OR = 1.9, 95% CI 1.0-3.7). After adjustment for body mass index, the presence of Heberden's nodes, previous hip injury and past leisure sporting activity (all independent risk factors for hip OA), and for other reproductive variables, there was a non-significant, protective effect of long-term hormone replacement therapy, such that > or =5 yr of use was associated with a 40% reduction in risk (OR = 0.6, 95% CI 0.2-1.8). Paradoxically, short term HRT use (up to 5 yr duration) was associated with an excess risk of hip OA (OR = 1.7, 95% CI 0.9-3.3). There was no association between the risk of hip OA and use of oral contraceptives, parity or hysterectomy. CONCLUSIONS: These data are consistent with previous studies suggesting a protective effect of long-term hormone replacement therapy on the risk of hip OA. By contrast, an elevation of risk in short-term users was demonstrated. Our results also suggest that risk is increased among women who have undergone unilateral or bilateral oophorectomy. Studies are required to investigate the mechanisms underlying these associations. PMID- 9851270 TI - Tumour necrosis factor microsatellites in reactive arthritis. AB - The purpose was to study tumour necrosis factor (TNF)-a, -b and -c microsatellites as potential new susceptibility markers for reactive arthritis (ReA). Fifty-nine patients typed for HLA-B27 were studied for frequencies of TNF microsatellite alleles and compared with allele frequencies determined from 285 random haplotypes and 46 healthy HLA-B27-positive controls. TNFa, -b and -c microsatellite sequences were amplified by the polymerase chain reaction, and the size of the product was defined by an automated sequencer. The frequencies of TNFa6 and -c1 alleles were found to be increased in patients with ReA, whereas TNFa11 and -c2 frequencies were decreased as compared to control haplotypes. The increase in the c1 allele in patients with ReA independently from HLA-B27 suggests that it might be a new susceptibility marker for the disease. The association of ReA with other alleles was due to a linkage disequilibrium with HLA-B27. PMID- 9851271 TI - Interleukin-10 inhibits the capacity of synovial macrophages to function as antigen-presenting cells. AB - OBJECTIVE: We have investigated the effects of interleukin (IL)-10, IL-4 + granulocyte/macrophage colony-stimulating factor (GM-CSF) and tumour necrosis factor alpha (TNF-alpha) on the phenotype and antigen-presenting capacity of synovial fluid (SF) macrophages from patients with rheumatoid arthritis. METHODS: The effects of IL-4, IL-10, GM-CSF and TNF-alpha on the expression of surface antigens on SF macrophages were studied using flow cytometry. The effects of these cytokines on the capacity of SF macrophages to activate T cells was investigated using the allogeneic mixed lymphocyte reaction (MLR). RESULTS: IL-10 reduced the expression of CD40, CD86 and HLA-DR, and increased the expression of CD14, on SF macrophages. IL-10 had no effect on the expression of CD80. Importantly, these effects of IL-10 on the phenotype of SF macrophages appear to have functional consequences, because cells incubated with IL-10 had a significantly reduced capacity to activate T cells in MLR. The effects of IL-4, GM-CSF and TNF-alpha were generally opposite to those observed in response to IL 10. IL-4 + GM-CSF, a combination of cytokines known to induce differentiation of dendritic cells, increased the expression of CD40, CD80 and CD86, and decreased the expression of CD14 on SF macrophages. Accordingly, IL-4 + GM-CSF increased the capacity of SF macrophages to activate T cells in MLR. IL-10 inhibited the effects of IL-4 + GM-CSF on SF macrophages. CONCLUSIONS: IL-10 inhibits the antigen-presenting capacity of SF macrophages, which further emphasizes the anti inflammatory potential of IL-10 in RA. Importantly, IL-10 is able to downregulate the APC function of SF macrophages even when they are efficiently activated. PMID- 9851272 TI - Referral and diagnosis of common rheumatic diseases by primary care physicians. AB - OBJECTIVE: To describe primary care patterns of referral and diagnoses of patients with rheumatic diseases referred to rheumatologists. METHODS: The medical records of all consecutive patients referred in 1994 by >300 primary care physicians to two rheumatologists at an academic centre were reviewed. The referring physician diagnosis was compared with the rheumatologist's diagnosis. Sensitivity, specificity and predictive values of primary care diagnoses were estimated using the rheumatologist diagnosis as the 'gold standard'. SETTING: University-based rheumatology out-patient clinic. RESULTS: Over half of the patients referred had a rheumatologist diagnosis of soft-tissue rheumatism or a spinal pain syndrome. Three hundred and forty-seven patients (49%) had a primary care diagnosis of a defined rheumatic disease. Of these, 142 (41%) of the primary care diagnoses were subsequently modified by the rheumatologist. The highest agreement between primary care physician and rheumatologist was observed for crystal-induced arthritis (kappa = 0.86), and the lowest agreement for polymyalgia rheumatica (kappa = 0.39) and systemic lupus (kappa = 0.46). Sensitivity was lowest for a primary care diagnosis of fibromyalgia (48%) and highest for ankylosing spondylitis (94%). Positive predictive values were generally low, in particular for systemic lupus erythematosus (33%) and polymyalgia rheumatica (30%). CONCLUSION: Most patients referred to an academic rheumatology centre had soft-tissue rheumatism or other pain syndromes. In general, diagnostic agreement between rheumatologists and primary care physicians was low. Increased emphasis on musculoskeletal disorders should be encouraged in medical education to increase the efficiency of rheumatology referrals. PMID- 9851273 TI - Progression in early erosive rheumatoid arthritis: 12 month results from a randomized controlled trial comparing methotrexate and gold sodium thiomalate. AB - OBJECTIVE: To compare radiographic outcomes in patients with active early erosive rheumatoid arthritis (RA) who were treated with methotrexate (MTX) and gold sodium thiomalate (GSTM). METHODS: A total of 174 patients from two centres were randomly assigned to receive weekly i.m. injections for 12 months of either 15 mg MTX or 50 mg GSTM in a double-blind fashion. Radiographic evaluations including standardized scoring of 38 joints of the hands, wrists and forefeet, and count of eroded joints, were carried out at baseline and after 6 and 12 months in all patients, including withdrawals. RESULTS: An intention-to-treat analysis revealed no statistically significant difference in the progression of radiographic scores between treatment groups after 6 months (3.4 with MTX vs 2.6 with GSTM, P = 0.66) and after 12 months (6.0 vs 4.8, P = 0.44). A similar pattern was observed for the number of joints with erosions. The slope of radiographic progression was significantly reduced in the second half-year compared to the first 6 months in both groups. Erythrocyte sedimentation rate and C-reactive protein at baseline, and the presence of rheumatoid factor (RF), were the main predictors of progression in bivariate analysis. RF remained as the only predictor for radiographic outcome in multivariable analysis. CONCLUSION: In parallel to clinical improvement, both GSTM and MTX reduce the slope of radiographic progression in patients with active erosive RA. PMID- 9851274 TI - Clinical implications of soluble intercellular adhesion molecule-1 levels in systemic sclerosis. PMID- 9851275 TI - Multiple antiphospholipid tests do not increase the diagnostic yield in antiphospholipid syndrome. AB - The family of antiphospholipid antibodies (aPL) includes a heterogeneous population of autoantibodies whose specificity is directed against not only phospholipids, but their complex with plasma proteins. Anticardiolipin antibodies (aCL) and lupus anticoagulant (LA) tests are widely performed to screen the aPL family which is associated with thrombotic complications in patients with systemic lupus erythematosus (SLE) or antiphospholipid syndrome (APS). The clinical significance of other aPL tests, including antibodies against phosphatidylserine (aPS), phosphatidylinositol (aPI), phosphatidic acid (aPA), phosphatidylcholine (aPC) and phosphatidylethanolamine (aPE), has not been established. The purpose of this study was to evaluate whether multiple aPL tests have enhanced diagnostic value for APS. We tested IgG/M/A aPS, aPI, aPA, aPC and aPE by ELISA using 10% bovine serum as blocking and sample diluent in 26 SLE patients with clinical manifestations of APS, but negative for both aCL and LA (Group 1). The results were compared with 32 SLE patients without any features of APS (Group 2) and 24 SLE patients with APS (aCL and/or LA positive) (Group 3). In Group 1, 1/26 (4%) was positive for IgA aPE, less frequent than in other groups, and none of the patients had any other aPL. In Group 2, 1/32 (3%) was positive for aPS, two (6%) for aPI, one (3%) for aPA and four (12.5%) for aPE. None was positive for aPC. In the third group, 13/24 (54%) were positive for aPS, 11 (46%) for aPI, 15 (63%) for aPA, four (17%) for aPC and seven (29%) for aPE. Since aPE was found in some patients, we extended the study, including 207 SLE patients, and tested aPE. IgG/M/A aPE was found in six (3%), 10(5%) and 21 (10%), respectively, but no association was found between aPE and any clinical features of APS. This study suggests that screening by multiple aPL tests does not increase the diagnostic yield in APS. PMID- 9851276 TI - An insight into rheumatology resources available on the World Wide Web. AB - The aim of this study was to gain an overview of rheumatology resources on the World Wide Web (WWW). A list of websites was generated using a commercial search engine and 'rheumatology' as a key word. A total of 154 websites were then evaluated with respect to origin and likely target audience; 43% of this initial group were either not accessible, repeats, or in a language other than English. Of the 87 websites we were able to analyse, we found that 67% originated from medical organizations and 51% were interpreted to be directed specifically at rheumatologists. Only 16% of websites were directed at patients only. The remainder were felt to contain information useful to both groups. Over half the websites felt to be of interest to patients contained advertisements. Although there is a lot of information relating to rheumatology on the WWW, it was invariably time consuming to access and there was little directed solely at patient education. PMID- 9851277 TI - Systemic sclerosis complicated by procainamide-induced lupus and antiphospholipid syndrome. PMID- 9851278 TI - The use of intravenous pulsed methylprednisolone in the treatment of systemic onset juvenile chronic arthritis. AB - An open prospective study using i.v. methylprednisolone in children with juvenile chronic arthritis (JCA) who had had a systemic exacerbation of disease is described. Eighteen children aged from 3 to 14 yr and 9 months (mean 9.7 yr) were treated. Ten patients (55%) had a loss of all systemic features 1 month after the pulse, and eight (45%) had a reduction in the active joint count. At this time, five of the patients on oral prednisolone had achieved a reduction in dosage. Also at 1 month, a reduction in erythrocyte sedimentation rate was observed in 11 patients (61%) and of C-reactive protein in 11 of 16 (72%). Altogether, 13 patients (72%) had a good response, while a further three (16%) went into remission. Our conclusions are that pulse methylprednisolone provides good short term benefit in patients with systemic-onset JCA; no serious side-effects were noted. Further long-term studies are warranted. PMID- 9851280 TI - George Frederic Still--registrar, Great Ormond Street Children's Hospital. PMID- 9851279 TI - Monoarticular chronic synovitis in a child. AB - Lipoma arborescens is a villous lipomatous proliferation of the synovial membrane characterized by chronic and painless synovial effusion. The aetiology is unknown. It has to be included in the differential diagnosis of chronic monoarticular disease in childhood. Magnetic resonance imaging provides a highly efficient tool for the diagnosis of this very rare condition. This is indeed the fourth paediatric case reported. Rather than resorting to the often inconvenient surgical synovectomy commonly recommended, we chose to treat the knee of this 13 yr-old boy with intra-articular osmic acid. PMID- 9851281 TI - Unusual high frequency of multifocal lesions of osteonecrosis in a young patient with systemic lupus erythematosus. PMID- 9851282 TI - Septic arthritis due to Fusobacterium nucleatum. PMID- 9851283 TI - HLA antigens in familial Behcet's disease in Ireland. PMID- 9851284 TI - Risk factors for thrombotic events in giant cell arteritis and polymyalgia rheumatica. PMID- 9851285 TI - Re: Use of cyclic etidronate and the prevention of non-vertebral fractures. PMID- 9851287 TI - No serological evidence to implicate a role for cytomegalovirus infection in the aetiology of Felty's syndrome. PMID- 9851286 TI - Serum dehydroepiandrosterone sulphate levels in patients with early rheumatoid arthritis: positive association with C-reactive protein, but not with HLA-DR genotype. PMID- 9851288 TI - Growth inhibitory factor and zinc affect neural cell cultures in a tissue specific manner. AB - Deficiency of neuronal growth inhibitory factor (GIF) and abnormalities in zinc homeostasis have been suggested to play a role in the neuropathogenesis of Alzheimer's disease. We report here that embryonic chick cerebral cell cultures zinc and copper containing GIF in the presence of marmoset hippocampal extract reduces significantly and concentration dependently mitochondrial succinate dehydrogenase activity (MTT) and cell mass. In contrast, no indications could be found that GIF affected neural retina cell cultures. Our results suggest that the observed effects of GIF are not elicited by zinc. PMID- 9851289 TI - Structural determinants of cytochrome P450 substrate specificity, binding affinity and catalytic rate. AB - The structural characteristics of cytochrome P450 substrates are summarised, showing that molecular descriptors can discriminate between chemicals of differing P450 isozyme specificity. Procedures for the estimation of P450 substrate binding interaction energies and rates of metabolism are described, providing specific examples in both individual compounds binding to P450s, including those of known crystal structure, and within series of structurally related chemicals. It is demonstrated that binding energy components are primarily hydrophobic/desolvation and electrostatic/hydrogen-bonded in nature, whereas electronic factors are of importance in determining variations in reaction rates. It is thus shown that the prediction of P450 substrate binding affinities and catalytic rates may be feasible, provided that sufficient structural information is available for the relevant enzyme-substrate complex. PMID- 9851290 TI - Redox cycling of polycyclic aromatic hydrocarbon o-quinones: metal ion-catalyzed oxidation of catechols bypasses inhibition by superoxide dismutase. AB - Several two-electron quinone reductases catalyze the redox cycling of polycyclic aromatic hydrocarbon (PAH) o-quinones. When the carbonyl reductase of human placenta catalyzes the cycling of 9,10-phenanthrenequinone in aqueous phosphate buffer, reactive oxygen species are produced. Superoxide dismutase (SOD) inhibits the cycling by more than 90%, but the addition of 1 microM Cu2+ or 15 microM ferricytochrome c (cyt c3+) completely restores the cycling rate to that of the control. Similar results are obtained for 5,6-chrysenequinone, 5,6 benz[a]anthracenequinone, 4,5-benzo[a]pyrenequinone, and 7,8 benzo[a]pyrenequinone in assay mixtures which contain dimethyl sulfoxide. The 17beta-hydroxysteroid dehydrogenase (17beta-HSD) of human placenta also catalyzes the redox cycling of these quinones, and cycling is inhibited by SOD. Although free metal ions (Cu2+ and Fe3+) inhibit the 17beta-HSD, cyt c3+ does not inhibit the enzyme. If cyt c3+ is added to assay mixtures containing SOD, cycling rates are equal to those of the corresponding controls. These experiments suggest that SOD may not protect cells from the toxic effects of PAH o-quinone cycling if certain metal ions or metal chelates are also present. PMID- 9851291 TI - Cytostatic mechanism and antitumor potential of novel 1H-cyclopenta[b]benzofuran lignans isolated from Aglaia elliptica. AB - A total of five 1H-cyclopenta[b]benzofuran lignans (1-5) isolated from the stems of Aglaia elliptica B1. (Meliaceae) inhibited the growth of human cancer cells in culture. Of particular note, the IC50 values observed with 1 (methyl rocaglate), 2 (4'-demethoxy-3',4'-methylenedioxy-methyl rocaglate) and 5 (1-O-formyl-4' demethoxy-3',4'-methylenedioxy-methyl rocaglate) were in the 1-30 ng/ml range. Prompted by the high potency of these responses, additional studies were performed with 2, a structurally representative isolate that was available in sufficient quantity as a result of the isolation process. Utilizing cultured Lu1 (human lung carcinoma) cells as a model, compound 2 induced accumulation in the G1/G0 phase of the cell cycle after 24 or 32 h of incubation; normal cell-cycle dynamics were observed at subsequent time periods. Cell proliferation was inhibited in a dose-dependent manner, but during the course of wash-out experiments, colony formation was not reduced. In addition, as judged by [3H]leucine incorporation, the test compound strongly inhibited protein biosynthesis (IC50 = 25 ng/ml). In analogous studies, nucleic acid biosynthesis was not reduced, even when cells were treated with concentrations as high as 1 microg/ml. These data suggest inhibition of protein synthesis is a key mode of action, and the compound functions by a cytostatic mechanism. Utilizing a human breast cancer cell line (BC1) sensitive to compound 2 in culture (IC50 = 0.9 ng/ml), an initial assessment of antitumor potential was performed. In accord with the in vitro results, the growth of BC1 in athymic mice was delayed by treatment with compound 2 (10 mg/kg body weight, three times per week, i.p.). Body weight was unaffected and no signs of overt toxicity were observed. However, growth paralleled that of the control group at later time points. Thus, novel 1H cyclopenta[b]benzofuran lignans are potent cytostatic inhibitors of protein biosynthesis and are capable of delaying tumor growth in an in vivo model. Their full clinical or basic utility requires further investigation. PMID- 9851292 TI - Tissue distribution of DNA adducts in male Fischer rats exposed to 500 ppm of propylene oxide: quantitative analysis of 7-(2-hydroxypropyl)guanine by 32P postlabelling. AB - 7-(2-Hydroxypropyl)guanine (7-HPG) constitutes the major adduct from alkylation of DNA by the genotoxic carcinogen, propylene oxide. The levels of 7-HPG in DNA of various organs provides a relevant measure of tissue dose. 7-Alkylguanines can induce mutation through abasic sites formed from spontaneous depurination of the adduct. In the current study the formation of 7-HPG was investigated in male Fisher 344 rats exposed to 500 ppm of propylene oxide by inhalation for 6 h/day, 5 days/week, for up to 20 days. 7-HPG was analyzed using the 32P-postlabelling assay with anion-exchange cartridges for adduct enrichment. In animals sacrificed directly following 20 days of exposure, the adduct level was highest in the respiratory nasal epithelium (98.1 adducts per 10(6) nucleotides), followed by olfactory nasal epithelium (58.5), lung (16.3), lymphocytes (9.92), spleen (9.26), liver (4.64), and testis (2.95). The nasal cavity is the major target for tumor induction in the rat following inhalation. This finding is consistent with the major difference in adduct levels observed in nasal epithelium compared to other tissues. In rats sacrificed 3 days after cessation of exposure, the levels of 7-HPG in the aforementioned tissues had, on the average, decreased by about one-quarter of their initial concentrations. This degree of loss closely corresponds to the spontaneous rate of depurination for this adduct (t 1/2 = 120 h), and suggests a low efficiency of repair for 7-HPG in the rat. The postlabelling assay used had a detection limit of one to two adducts per 10(8) nucleotides, i.e. it is likely that this adduct could be analyzed in nasal tissues of rats exposed to less than 1 ppm of propylene oxide. PMID- 9851293 TI - Spectroscopy features of the binding of polyene antibiotics to human serum albumin. AB - The alteration in the fluorescence spectra observed for the polyene antibiotics nystatin and amphotericin B in the presence of human serum albumin is due to a decrease in the polar character of the antibiotic environment when these are bound to the protein. Amphotericin B showed two types of binding sites, the first having a very high affinity (5.8 x 10(7) M(-1)) and a secondary binding site with an affinity two orders lower than the primary site. This secondary binding site was very sensitive to temperature change. Nystatin yielded only one type of binding site with an affinity of 1.1 x 10(5) M(-1). Nystatin was found to be bound to fatty acid binding sites in albumin, while amphotericin B was not, suggesting that the fatty acid binding sites are not simple, depending on the number of unsaturated bonds on the polyene antibiotic molecule. Both polyene antibiotics displaced bilirubin bound to albumin, which is in agreement with the similarities of the affinity values of this chromophore and the polyene antibiotics with albumin. PMID- 9851294 TI - Reduced baroreflex sensitivity and cardiorespiratory transfer in amyotrophic lateral sclerosis. AB - INTRODUCTION: Clinically relevant autonomic disturbances have been reported for respirator-dependent ALS patients while subclinical involvement may be present in the early course. METHODS: Eighteen patients with early-stage ALS and 18 age matched controls were studied by means of standard autonomic tests (heart off + response to deep breathing and tilt-table testing), and spectral analysis of heart rate (HR) and arterial blood pressure (ABP), using the associated transfer function as a measure of baroreflex sensitivity for the mid-frequency band (MF band, 0.05-0.15 Hz) and as a measure of cardiorespiratory transfer for the high frequency band (HF band, 0.15-0.33 Hz). RESULTS: Mean HR and ABP were increased in ALS, while results of standard autonomic tests were similar for ALS and controls. Transfer function analysis revealed reduced baroreflex sensitivity and diminished cardiorespiratory transfer during normal breathing. CONCLUSIONS: Cardiovascular autonomic functions are intact in patients with ALS. There is evidence of sympathetic enhancement and vagal withdrawal, accompanied by reduced baroreflex sensitivity. These findings are similar to those reported for essential hypertension and may point to a common central autonomic derangement in both disorders. PMID- 9851295 TI - Cortico-cortical inhibition of the motor cortical area projecting to sternocleidomastoid muscle in normals and patients with spasmodic torticollis or essential tremor. AB - OBJECTIVES: To investigate whether the cortico-cortical inhibition originally reported for the human hand motor area is present in the motor cortex for sternocleidomastoid muscle (SCM) and to evaluate the amount of inhibition in spasmodic torticollis and essential tremor. METHODS: Subjects were 14 normal healthy volunteers, 10 patients with spasmodic torticollis and 5 with essential tremor involving neck muscles. A paired-pulse magnetic stimulation was performed for the SCMs and first dorsal interosseous muscles (FDIs). RESULTS: In normal subjects, a subthreshold magnetic conditioning stimulus suppressed responses to a suprathreshold magnetic test stimulus when their interval was 1-5 ms in SCM. This indicates that the similar cortico-cortical inhibitory mechanism is present in the motor cortex for SCM as in the hand motor area. In the patients with spasmodic torticollis, the cortico-cortical inhibitory effect was reduced or absent in SCM, but normal in the FDI. In contrast, in patients with essential tremor, normal cortico-cortical inhibition was seen in both the SCM and FDI. CONCLUSIONS: The cortico-cortical inhibitory mechanisms of the motor cortex for SCM can be studied by a paired-pulse magnetic stimulation method. Our result of reduced cortico-cortical inhibition in torticollis patients suggests abnormal excitability (hyperexcitable or disinhibited) of the motor cortex for SCM in spasmodic torticollis. PMID- 9851296 TI - Comparison of descending volleys evoked by transcranial magnetic and electric stimulation in conscious humans. AB - OBJECTIVES: The present experiments were designed to compare the understanding of the transcranial electric and magnetic stimulation of the human motorcortex. METHODS: The spinal volleys evoked by single transcranial magnetic or electric stimulation over the cerebral motor cortex were recorded from a bipolar electrode inserted into the cervical epidural space of two conscious human subjects. These volleys were termed D- and I waves, according to their latency. Magnetic stimulation was performed with a figure-of-eight coil held over the right motor cortex at the optimum scalp position, in order to elicit motor responses in the contralateral FDI using two different orientations over the motor strip. The induced current flowed either in a postero-anterior or in a latero-medial direction. RESULTS: At active motor threshold intensity, the electric anodal stimulation evoked pure D activity. At this intensity, magnetic stimulation with the induced current flowing in a posterior-anterior direction evoked pure I1 activity. When a latero-medial induced current was used, magnetic stimulation evoked both D and I1 activity. Using electric anodal stimulation, at a stimulus intensity of 9% of the stimulator output above the active motor threshold (corresponding approximately to 1.5 active motor threshold), a small I1 wave appeared only in subject 1. Using magnetic stimulation with a posterior-anterior induced current, at a stimulus intensity of 21% of maximum stimulator output above the active motor threshold (corresponding approximately to 1.8 times threshold in subject 1 and to two times threshold in subject 2), a small D wave appeared in subject 1 but not in subject 2. CONCLUSIONS: Present results demonstrate that, in conscious humans at threshold intensities, electric stimulation evokes D waves and magnetic stimulation (with a posterior-anterior induced current) evokes I waves, while magnetic stimulation (with a latero-medial induced current) evokes both activities. PMID- 9851297 TI - Postural reflexes in patients with HIV-1 infection. AB - OBJECTIVE: We studied whether medium latency (ML) and long latency (LL) postural reflexes, which are abnormal in a number of neurological conditions including basal ganglia disorders, provide an early marker of CNS involvement in HIV positive patients. METHODS: Leg reflexes were elicited in 9 neurologically normal HIV-positive patients and 10 healthy controls who were standing upright, using toe-up forceplate rotations of varying amplitude (4 degrees and 10 degrees) and predictability (serial and random). RESULTS: For predictable amplitude perturbations, posturally destabilizing ML and stabilizing LL responses in HIV seropositives did not differ from controls. However, for unpredictable amplitude perturbations, HIV-positive patients inappropriately manifested a mid-size default LL response, in contrast to healthy subjects who showed a maximum size default response. CONCLUSIONS: These results suggest that impaired modulation of LL reflex processing occurs in early stages of HIV infection, prior to the onset of clinical postural instability, and this dysregulation may be influenced by cognitive factors. PMID- 9851298 TI - Facilitation of motor evoked potentials (MEPs) in first dorsal interosseous (FDI) muscle is dependent on different motor images. AB - OBJECTIVE: We investigated changes in motor evoked potentials (MEPs) to explain why mental practice can improve motor performance. METHODS: MEPs were recorded from right and left first dorsal interosseous (FDI) muscles of 9 normal, right handed subjects during different motor images of index finger movement: (1) rest, (2) flexion, (3) abduction, (4) extension. A paired t test was used to compare differences of stimulus intensities and MEP amplitudes among conditions. RESULTS: MEP amplitudes significantly increased in both FDI muscles during motor images of flexion and abduction but not of extension. Moreover, MEP amplitudes were larger in flexion than in abduction. These differences were proportional to the amount of real EMG discharge of FDI muscle in the selected direction of index finger movement. With regard to right-left differences, MEP amplitudes in the right FDI muscle were larger than those in the left. CONCLUSIONS: The primary motor cortex plays a role in the mental representation of motor acts. Furthermore, the amount of corticomotoneuronal cell activity is affected by the different motor images utilizing the same muscle. Right-left difference of MEP amplitude supports the view of left-hemisphere dominance for motor programming as an aspect of normal brain function among right-handers. PMID- 9851299 TI - Clinical and electrophysiological findings and follow-up in tarsal tunnel syndrome. AB - The authors report clinical and electrophysiological findings in 59 patients with tarsal tunnel syndrome (TTS) and follow-up in 23 of them. The entrapment was prevalent in females; was bilateral in 6 patients and involved medial plantar in 7 and lateral plantar nerves in two cases. Eleven presented with other nerve entrapment syndromes or focal mononeuropathies, due to hereditary neuropathy with liability to pressure palsy or systemic diseases. The other 48 subjects had TTS without any other related entrapment syndromes: 23 were idiopathic cases, 13 had a history of local trauma, 3 had systemic diseases and the others had external or intrinsic compressions. The most frequent symptoms were paraesthesia or dysaesthesia (86% of feet) and pain (55%). Hypoaesthesia of the sole and weakness of toe flexion were evident in 74% and 22% of feet, respectively. Absence of sensory action potential or slowing of sensory conduction velocity (SCV) of the plantar nerves were present in 77% of feet; significant differences of SCV between affected and unaffected plantar nerves and/or between distal sural and plantar nerves were evident in 14%. Abnormalities of plantar SCV were therefore absent in only 9% of feet. Distal motor latency was delayed in 55% and electromyography showed neurogenic changes in 45% of sole muscles. Five cases (6 feet) underwent surgery with excellent or good results in 5, 4 of them also showing improvement in distal conduction of the plantar nerves. Nine were treated with local steroid injections, with good results shown in 6 patients. Nine other patients who did not receive any therapy showed a disappearance of symptoms or good outcome in 6 cases. The subjects with poor therapeutic results had S1 radiculopathy or systemic diseases. The authors underline that patients with connective tissue diseases should not be treated by surgical decompression because they may have subclinical neuropathy. Some subjects with idiopathic or trauma-induced TTS recover spontaneously. Surgical release should be limited to cases with space-occupying lesions and when conservative treatments fail. PMID- 9851300 TI - Dependence of the transcranially induced silent period on the 'instruction set' and the individual reaction time. AB - OBJECTIVES AND METHODS: We looked for influences of the experimental condition on the silent period (SP) from transcranial motor cortex stimulation and analyzed how the instruction given to the subject, as well as the individual reaction time, might affect the duration of the SP in the biceps brachii muscle. RESULTS: The duration of the SP was found to critically depend on the subject's voluntary reaction of the target muscle immediately after the stimulus. With low stimulus intensity and low background force, the duration of the silent period was significantly longer in 10 of 13 subjects (P = 0.002) when they were instructed to relax quickly after the stimulus rather than to maintain the the force at a constant level. A significant shortening of the SP (P = 0.02) was observed when the subjects were instructed to perform a rapid contraction of the target muscle in reaction to the cortical stimulus. With low stimulus intensity and high background force, the same influence of the instruction set was found in 6 of 13 subjects. When the subjects were left without precise instruction, the SP duration was unpredictable. In 10 subjects, the SP corresponded to that obtained with the instruction to maintain the force at a constant level. However, in 3 subjects it was prolonged to the value observed in the 'relax' instruction. With greater stimulus intensities, the effect of the instruction set on the SP duration was generally smaller. A significant prolongation was nevertheless found at low background forces with rapid relaxation (P < 0.001), and a significant shortening was found at high background forces with rapid contraction (P < 0.001) after the stimulus. The SP duration observed with 20% of maximal voluntary contraction (MVC) significantly correlated with the individual reaction time. No such correlation was found for the SP obtained with 80% MVC. The SP was slightly longer at 20% MVC, as compared to 80% MVC within each instruction group. This effect was significant (P < 0.05) at low stimulus intensities. CONCLUSIONS: Therefore, when assessing the SP duration for diagnostic purposes, not only the stimulus intensity but also the background force and the voluntary reaction must be standardized. Furthermore, great stimulus intensities and high background forces should be used to minimise the effects of instruction set and individual reaction time. PMID- 9851301 TI - Relation between the size of motor units and the spectral characteristics of their action potentials. kazuseki@iname.com. AB - OBJECTIVE: The purpose of this study was to clarify the influence of the size of an individual motor unit on the spectral characteristics of its motor unit action potentials (MUAPs). METHODS: In 4 human subjects, we first averaged the isometric force and the surface EMG signals triggered by the intramuscularly recorded action potentials of each motor unit during voluntary isometric contraction. Then, we obtained averaged twitch contraction curves, averaged MUAPs derived from surface EMG signals (S-MUAPs), and the power spectrum of the S-MUAPs. Finally, we tested for correlations among these results in each subject. RESULTS: There was a positive correlation among the mean power frequency of the S-MUAPs, the maximal amplitude of the S-MUAPs, and the maximal twitch tension of their motor unit in each subject (r = 0.58-0.85; P < 0.05). CONCLUSIONS: Since the twitch tension and the amplitude of the S-MUAPs are well-known predictors of the size of motor units, we conclude that the spectral characteristics of the S-MUAPs reflect the size of the motor unit from which they originate. PMID- 9851302 TI - Cortical activation during fast repetitive finger movements in humans: steady state movement-related magnetic fields and their cortical generators. AB - OBJECTIVE: To study the cortical physiology of fast repetitive finger movements. METHODS: We recorded steady-state movement-related magnetic fields (ssMRMFs) associated with self-paced, repetitive, 2-Hz finger movements in a 122-channel whole-head magnetometer. The ssMRMF generators were determined by equivalent current dipole (ECD) modeling and co-registered with anatomical magnetic resonance images (MRIs). RESULTS: Two major ssMRMF components occurred in proximity to EMG onset: a motor field (MF) peaking at 37+/-11 ms after EMG onset, and a postmovement field (post-MF), with inverse polarity, peaking at 102+/-13 ms after EMG onset. The ECD for the MF was located in the primary motor cortex (M1), and the ECD for the post-MF in the primary somatosensory cortex (S1). The MF was probably closely related to the generation of corticospinal volleys, whereas the post-MF most likely represented reafferent feedback processing. CONCLUSIONS: The present data offer further evidence that the main phasic changes of cortical activity occur in direct proximity to repetitive EMG bursts in the contralateral M1 and S1. They complement previous electroencephalography (EEG) findings on steady-state movement-related cortical potentials (ssMRCPs) by providing more precise anatomical information, and thereby enhance the potential value of ssMRCPs and ssMRMFs for studying human sensorimotor cortex activation non invasively and with high temporal resolution. PMID- 9851303 TI - Human somatosensory evoked magnetic fields to vibratory stimulation of the index finger: is there frequency organization in SI? AB - OBJECTIVE: Frequency organization in the human somatosensory cortex was studied. DESIGN AND METHODS: Somatosensory evoked magnetic fields (SEFs) from 12 subjects were measured following vibratory stimulation of the index finger by using a 122 channel whole head SQUID system. Sensory stimuli comprising a 40 ms vibration at frequencies of 50, 100, 200 and 400 Hz were delivered to the volar surface of the tip of the right index finger. Using a single dipole model, the sources of the magnetic fields were estimated and mapped onto magnetic resonance images of each subject. The analysis of variance test (ANOVA) was used for statistics. RESULTS: Source localization was determined on the main two peaks (M60 and M110) of the SEFs. All of the sources were located in the area 3b of somatosensory cortex (SI). There were no statistically significant differences between the locations of the dipoles evoked by different frequency stimulations. CONCLUSIONS: These results demonstrate the absence of systematic frequency organization at the hand representation area of the SI cortex. We speculate that high frequency vibration above 100 Hz are coded by the fast-spiking interneurons which synapse with Pacinian pyramidal neurons in SI. PMID- 9851304 TI - The physiological origin of the slow afterwave in muscle action potentials. AB - OBJECTIVE: Both intramuscularly-recorded motor unit action potentials (MUAPs) and surface recorded MUAPs and compound muscle action potentials (CMAPs) have slow afterwaves which can contribute as much as half their measured duration. This study tested the hypothesis that the slow afterwave has its physiological origin in the negative afterpotential of the muscle fiber intracellular action potential (IAP). METHODS: We investigated the slow afterwave in MUAPs and CMAPs from brachial biceps, tibialis anterior, first dorsal interosseous, thenar and hypothenar muscles in 15 normal subjects, and using computer simulations. RESULTS: The slow afterwaves did not match the time constant of the amplifier's high-pass filter, and so were not filtering artifacts. They lasted long after propagation had terminated at the muscle/tendon junction, and so were not due to the temporal or spatial dispersion of propagating single-fiber potentials. Their amplitude and polarity varied with the recording site as predicted by computer simulations that modeled the IAP as having a negative afterpotential. They also changed with double-pulse stimulation and decreasing temperature in ways consistent with the results of intracellular studies of the IAP negative afterpotential. CONCLUSIONS: The presented results support our hypothesis that the slow afterwave is a manifestation of the IAP negative afterpotential. PMID- 9851305 TI - Enzyme evolution and cancer: hypothesis why natural carcinogens are more potent than synthetic ones. AB - A great deal of evidence shows that carcinogen induced mutations in human cancers point towards natural rather than man-made agents. Here, we propose a model based on the premise that the evolutionary pressure of nature renders natural carcinogens more potent than artificial ones, present in equal concentration, by suitably modifying kinetic parameters of carcinogen metabolizing enzymes. Enzymes are evolved to bind the transition state of substrates more strongly than substrates themselves, thus obtaining more elevate values of the specificity constant kcat/Km (Ksp). Natural selection optimizing the catalytic power at the proper substrate concentration by suitable raising the Km values, reduce the Gibbs standard activation energy (G(0#)), accelerating the conversion of natural precarcinogens to potent carcinogens. Conversely, "man-made" carcinogens, since the last century in the biosphere, are converted to active metabolites at a lower rate than natural chemicals and the slower rate of activation would allow protective enzymes and DNA repair machinery more time to clean up the damage. PMID- 9851306 TI - Ex vivo occupancy by tamsulosin of alpha1-adrenoceptors in rat tissues in relation to the plasma concentration. AB - At 0.5-12 h after oral administration of tamsulosin (2.3 micromol/kg) in rats, there was a significant decrease in specific [3H]prazosin binding in the prostate as compared to the control value. The greater decrease occurred in the submaxillary gland. The effect of tamsulosin was mainly due to a marked reduction of [3H]prazosin binding sites (Bmax) rather than to an increase in the dissociation constant (Kd). In contrast, there was only a slight decrease or no change in the [3H]prazosin binding in the spleen, heart, and cerebral cortex of tamsulosin-administered rats at 0.5-12 h. Oral administration of terazosin (21.7 micromol/kg) significantly increased Kd values for [3H]prazosin binding with little effect on Bmax values in the rat prostate at 3 and 6 h. The greater increases in Kd values were observed in the submaxillary gland, spleen and heart at 0.5-12 h. Terazosin had a slight effect on Kd values for the cerebral cortical [3H]prazosin binding. Tamsulosin was absorbed rapidly after oral administration at a dose of 2.3 micromol/kg in rats, and at 6 h, plasma concentration decreased markedly to approximately one-twentieth of the 0.5 h peak level. alpha1 Adrenoceptor occupancy was estimated as a percentage of decrease in Bmax values for [3H]prazosin binding in tissues of tamsulosin-treated rats compared with control rats. The alpha1-adrenoceptor occupancy by tamsulosin in the prostate and submaxillary gland occurred rapidly in parallel with the rise in plasma concentration of tamsulosin, and lasted for over 12 h despite the marked decrease in plasma concentration. Consequently, it is suggested that tamsulosin produces more selective and sustained occupancy in vivo of alpha1-adrenoceptors in the submaxillary gland and prostate of rats than in other tissues. PMID- 9851307 TI - Analysis of the promoter of the luteinizing hormone/human chorionic gonadotropin receptor gene in neuroendocrine cells. AB - We investigated the molecular basis of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor gene transcription in immortalized alphaT3 gonadotropes, hypothalamic GT1-7 and hippocampal HN33p neurons. Nuclear run-on transcription, as well as transfection assays with fusion constructs of luciferase and the 5'-flanking region of LH/hCG receptor gene, revealed that GT1 7 neurons transcribe more than the alphaT3 or HN33p cells. Transient transfection of truncated reporter gene constructs and gel mobility shift assays revealed that while all neuroendocrine cells use the same promoter, they contain different levels of promoter binding proteins. Higher levels of these proteins may explain increased transcription of the LH/hCG receptor gene in GT1-7 neurons compared with alphaT3 and HN33p cells. PMID- 9851308 TI - Differential effects of acute morphine administrations on polymorphonuclear cell metabolism in various mouse strains. AB - This paper shows that an acute morphine treatment dose-dependently alters the energetic and oxidative metabolism of polymorphonuclear leukocytes obtained from BALB/c and DBA/2 mice, while phagocytic cells from C57BL/6 were not affected. In sensitive mouse strains, i.e. BALB/c and DBA/2, morphine decreased both ATP concentration and energy charge potential. At the same time, ATP catabolic products, i.e. nucleosides (inosine+adenosine) and oxypurines (hypoxanthine+xanthine+uric acid), significantly increased, indicating an imbalance between energy production and consumption. Morphine treatment also induced malondialdehyde and superoxide anions production in leukocyte cells from sensitive mice. The opiate antagonist naloxone blocked morphine-induced modifications by the lower morphine dose. The same parameters in cells from C57BL/6 mice were not affected. These findings confirm that: i) the phagocytic cells are an important target for the in vivo effects of morphine, and ii) the genotype-dependent variation influences the immunological responsiveness to opiates. PMID- 9851309 TI - Supersensitivity of spinal opioid receptors to antagonists in intrathecal butorphanol and morphine dependence. AB - The purpose of this investigation was to evaluate changes in the sensitivity of spinal opioid receptors to selective antagonists in rats rendered dependent on intrathecal (i.t.) butorphanol and morphine. Using quantitative autoradiography, competitive binding assays with selective opioid antagonists were performed in the spinal cord sections of i.t. butorphanol- and morphine-dependent rats in which withdrawal was precipitated by i.t. naloxone. In butorphanol-dependent rats, the spinal kappa-opioid receptor developed a greater degree of antagonist supersensitivity than the spinal delta- and mu-opioid receptors did. In contrast, the spinal mu-opioid receptor became more sensitive than the delta-opioid receptor in morphine-dependent rats. These results indicate that differential supersensitivity of spinal opioid receptors was induced after chronic i.t. infusions of butorphanol and morphine. PMID- 9851310 TI - Effect of the CB1 receptor antagonist, SR141716A, on cannabinoid-induced ocular hypotension in normotensive rabbits. AB - The present study attempts to indirectly determine if a neuronal cannabinoid (CB1) receptor mediates the intraocular pressure (IOP) reduction effects of arachidonoyl ethanolamide (AEA), its R-alpha-isopropyl analog, and the non classical cannabinoid, CP-55,940. A series of these cannabinoids were dissolved in an aqueous 10-20% 2-hydroxypropyl-beta-cyclodextrin (2-HP-beta-CD) solution (containing 3% polyvinyl alcohol) and administered (25-62.5 microg) unilaterally to normotensive rabbit eyes. This was repeated on animals pre-treated with a subcutaneous injection (2.5 mg/kg) of the highly specific CB1 receptor antagonist, SR 141716A, dissolved in an aqueous 42% 2-HP-beta-CD solution. AEA, its R-alpha-isopropyl analog, and CP-55,940 reduced IOP upon topical application to a greater degree than was detected in the untreated eye. This reduction was eliminated for the latter two compounds by subcutaneous (s.c.) pretreatment of the rabbits with the CB1 receptor antagonist, but the IOP properties of AEA remained unchanged. SR 141716A administered alone (s.c.), elevated the IOP of both eyes. A CB1 receptor seems involved in the IOP reduction induced by either R alpha-isopropyl anandamide or CP-55,940. However, AEA apparently functions through a different mechanism. PMID- 9851311 TI - Chronopharmacological effects on nicotine repeated administration on heart rate, body temperature and locomotor activity circadian rhythms in rats. AB - The aim of this study was to compare morning and evening repeated nicotine administration on the circadian rhythms of heart rate (H), body temperature (T) and locomotor activity (A) in unrestrained rats by using implanted radio telemetry transmitters. The study was divided into three 7-day periods: a control period (P1), a treatment period (P2) and a recovery period (P3). During P2, four rats received nicotine (1mg.kg(-1)) subcutaneously at 09.00 h and four rats received nicotine in the same conditions at 21.00 h. For P1, P2 and P3, a power spectrum analysis was applied in order to determine the dominant period of rhythmicity. If H, T and A circadian rhythms were detected, the characteristics of these rhythms were determined by cosinor analysis, expressed as means+/-SEM and compared by ANOVA. Our results indicated: (1) a lack of detection of A circadian rhythm during P2 for the morning group while H and T circadian rhythms were detected for the morning and evening group whatever the period. (2) alterations of mesors, amplitudes and acrophases of H and T circadian rhythms for the morning and evening group during P2 and alterations of mesor, amplitude and acrophase of A circadian rhythm for the evening group. Furthermore these alterations were significantly different for the morning and evening group during P2. These results showed that the time of administration of nicotine differently affect H, T and A rhythms. The authors suggest that these effects can be mediated by central cholinergic and/or monoaminergic mechanisms. PMID- 9851312 TI - Inhibition of adenylate cyclase of rat hepatic membranes by glycosaminoglycans. AB - Glycosaminoglycans are long non-branched polysaccharides composed of repeating disaccharide units. In a previous in vitro study we have shown that such molecules are able to modulate substrate phosphorylation catalyzed by cAMP dependent protein kinase. Here, we investigate the impact of glycosaminoglycans, such as heparan sulfate, dermatan sulfate, chondroitin 4- and 6-sulfate, keratan sulfate and hyaluronic acid upon adenylate cyclase, which directly regulates cAMP dependent protein kinase activity via cAMP synthesis. In rat liver plasma membrane preparation we have determined forskolin- and guanosine-5'-beta, gamma imidotriphosphate-induced cAMP formation catalyzed by adenylate cyclase in the presence of increasing concentrations of glycosaminoglycans. The results indicate that glycosaminoglycans strongly influence enzymic conversion of ATP into cAMP. The highest reduction of adenylate cyclase activity is observed in the presence of dermatan sulfate and heparan sulfate. Moreover, the inhibitory effect of these two glycosaminoglycans is higher when guanosine-5'-beta, gamma-imidotriphosphate, instead of forskolin, is used as stimulator of adenylate cyclase. Further characterization of enzyme inhibition mediated by dermatan sulfate shows that this molecule exerts an inhibitory effect of mixed type. PMID- 9851313 TI - Characterization of ethyl (3-quinuclidinyl) acetate (EQA) as a ligand for acetylcholine receptors. AB - Recent studies suggest that neuronal nicotinic acetylcholine receptors (nAChRs) may play a role in several CNS disorders and that subtype selective nicotinic ligands may be useful in the treatment of these disorders. Ethyl (3 quinuclidinyl)acetate (EQA) is a bulky, reverse-ester analog of ACh, that produces signs of cholinergic stimulation that may be nicotinic in origin The objective of the present study was to further evaluate EQA as a potential cholinergic ligand. Behavioral studies, smooth muscle assays, and radioligand binding assays were performed on this novel ligand. The effects of EQA on blood pressure and acetylcholinesterase activity were also evaluated. The results of the study suggest that EQA is an agonist at peripheral nicotinic acetylcholine receptors and may have antagonist properties at central nicotinic receptors. PMID- 9851314 TI - Adrenomedullin regulation by thyroid hormone in the rat. AB - To investigate the effect of thyroid hormone on adrenomedullin (AM) changes during hyper- and hypothyroid states, we evaluated plasma AM concentrations and AM mRNA levels in lung tissue from hyper- and hypothyroid rats. AM peptide concentrations were significantly higher in plasma from hyperthyroid rats and lower in plasma from hypothyroid rats compared with control rats. AM mRNA transcripts were significantly increased in lung tissue from hyperthyroid rats and significantly decreased in lung tissue from hypothyroid rats compared with normal rats. These changes are paralleled alterations in AM production in cultured rat vascular smooth muscle cells, in which AM is regulated at the transcriptional level by thyroid hormone. Thus, thyroid hormone positively regulates AM production in vivo. PMID- 9851315 TI - Hypotension does not alter the antinociceptive effect of nifedipine. AB - We explored the relationship between antinociceptive and hypotensive effects of nifedipine (NIF) injected intraperitoneally ( ip, 15 mg/kg) and epidurally (epi, 20 microM), as compared to verapamil (VER, 10 mg/kg ip) and nitroglycerin (NTG, 0.1 and 0.15 mg/kg ip). The systolic blood pressure (BP) and tail-flick (TF) latencies were measured simultaneously every 10 min for 2 hours and individual values of both measurements were correlated. The highest antinociceptive as well as hypotensive effects were both measured in the group receiving NIF epi., with the correlation coefficient r2=0.2878. Injected ip., NIF revealed similar antinociceptive effect, whereas the other studied drugs were not effective. As to the degree of hypotensive activity, NIF epi was followed by VER, NTG 0.1, NIF ip. and NTG 0.15. No significant correlation was found between BP and TF latencies in any group receiving the drugs. We concluded that the antinociceptive response, measured by the tail-flick technique, is independent of the hypotensive activity of the studied drugs, including NIF. PMID- 9851316 TI - mRNAs coding for the calcium-sensing receptor along the rat nephron: effect of a low-phosphate diet. AB - We investigated the localization of mRNA encoding the calcium-sensing receptor (CaSR) along the rat nephron. For this purpose, we combined microdissection of nephron segments and RT-PCR techniques. The results indicate that mRNA encoding rat CaSR is present in rat glomeruli and distal segments (medullary thick ascending limb, cortical thick ascending limb, distal convoluted tubule and cortical collecting duct), whereas it was not detected in proximal convoluted tubules or proximal straight tubules. We also studied whether the CaSR transcription in kidney cortex was modified in response to low dietary phosphate. No significant changes were detected. Given the fact that a low-phosphate diet increased Ca2+ excretion by more than 50-fold, the results suggest that if the CaSR regulates Ca2+ reabsorption, it does so through receptor occupancy by Ca2+ rather than by changes in receptor expression. PMID- 9851317 TI - Epidermal growth factor upregulates aminopeptidase N and 5'-nucleotidase in human glomerular mesangial cells. AB - Because epidermal growth factor (EGF) is likely to be released in the glomeruli during glomerular injury and mesangial cells possess specific receptors for EGF, we thought it to be of interest to examine whether this growth factor could influence the expression of ectoenzymes in cultured human mesangial cells. EGF stimulated 5'-nucleotidase and aminopeptidase N activities in intact human mesangial cells in a time- (24-72 h) and dose-dependent (0.1-50 ng ml(-1)) manner. Maximum stimulation represented 2.7- and 2-fold basal activities for 5' nucleotidase and aminopeptidase N, respectively. EGF did not influence cyclic AMP production, and its effect on 5'-nucleotidase was additive to that of forskolin or 8-bromo-cAMP. In contrast, genistein (10 mg x ml(-1)), an inhibitor of tyrosine kinase, prevented EGF-dependent stimulation of aminopeptidase N and 5' nucleotidase, suggesting that protein phosphorylation was involved in the signaling mechanism. EGF stimulated specifically the latter two enzymes since it had no effect on other ectoenzymes including alkaline phosphodiesterase I and Mg2+-ATPase activities. These results demonstrate that EGF, via the control of 5' nucleotidase and aminopeptidase N, which are implied in adenosine formation and peptide processing, respectively, could play a role in human cultured mesangial cell contractility and proliferation. PMID- 9851318 TI - Osmotically induced conductance and capacitance changes in in vitro perfused rectal gland tubules of Squalus acanthias. AB - The rectal gland of Squalus acanthias is critically involved in the homeostasis of NaCl and water metabolism and hence in overall osmoregulation. In the present study, we have examined the acute responses of rectal gland slices and in vitro perfused rectal gland tubule (RGT) cells to the exposure to dilute and hypertonic peritubule solutions. Five series were performed. (i) With changes in osmolality, Western blots to monitor tyrosine, threonine and serine phosphorylation in rectal gland slices did not reveal clear-cut changes in phosphorylation patterns. All other series were performed in in vitro perfused RGT. (ii) Relative cell volume was estimated by fura-2 fluorescence using the emission at the isosbestic excitation wavelength of 360 nm. Hypotonic solution (-100 mmol/l NaCl) reduced fura-2 fluorescence by 16% and hypertonic solution (+100 mmol/l NaCl) had the opposite effect (+12%). (iii) Transepithelial resistance was increased markedly by hypotonic solution, probably by cell swelling, and the opposite was seen with hypertonic solutions. (iv) Whole-cell patch clamp experiments indicated that hypotonic solution hyperpolarized the cells, and increased membrane conductance and membrane capacitance. The latter two changes correlated significantly with each other. Hypertonic solution had the opposite effect. (v) Measurements of the fura-2 fluorescence ratio (340/380 nm) revealed that hypotonic solution (-NaCl) increased cytosolic Ca2+ activtiy ([Ca2+]i). Hypertonic solution had no detectable effect on [Ca2+]i. These data indicate that RGT cells are swollen by removal of NaCl from the bath solution. This causes an increase in [Ca2+]i and a predominant increase in K+ conductance and hyperpolarization. Urea apparently permeates these cells quite well and its addition (+U) or its removal (-U) had only moderate osmotic effects. The removal of urea and replacement by mannitol produced effects similar to those seen with hypertonic NaCl solution. PMID- 9851319 TI - Analysis of the role of apoptosis and cell proliferation in renal cystic disorders. AB - Cell proliferation and apoptosis were studied in 8 patients with inherited polycystic kidney disease and in 34 patients with acquired cystic kidney conditions including solitary and multilocular cysts and segmental tubular dilation. Intact renal tissue of 20 surgically removed tumorous kidneys served as control. Cell proliferation and apoptosis were demonstrated by immunohistochemical and in situ end-labeling methods. The percentage of positively stained nuclei was calculated and statistically analyzed. Both apoptosis and cell proliferation were significantly higher (p<0.001) in polycystic kidney disease. The percentage of positively stained nuclei in the whole kidney tissue with acquired cysts did not differ from controls although cell proliferation was significantly higher (p<0.001) in cells lining the cysts. Apoptotic cells were rarely found in the cystic epithelium or were even absent in these cases. Our data indicate that while polycystic kidneys seem to be characterized by abnormal cell survival, acquired renal cysts have different behavior in which so far unknown intracellular changes are more likely to cause tubular distension probably through induced cell proliferation. PMID- 9851320 TI - Role of kinins in the renoprotective effect of angiotensin-converting enzyme inhibitors in experimental chronic renal failure. AB - The aim of this study was to investigate whether the renoprotective effect of angiotensin-converting enzyme inhibitors (ACEIs) following 5/6 renal mass reduction is due in part to the potentiation of kinins. Three groups of rats with 5/6 renal mass reduction were studied during the 14 weeks following surgery. One group received no therapy (control); the second group was treated from the beginning with the ACEI ramipril (1 mg/kg/day) added to the drinking water, and the last group received ramipril plus a beta2-bradykinin antagonist, HOE 140 (500 microg/kg/day) via osmotic minipumps. Plasma creatinine did not change in any group during the study. Urinary protein excretion rose in the controls from 9.18+/-1.6 to 45.0+/-5.6 mg/24 h at the end of the study. In ramipril group proteinuria was prevented (initial 7.5+/-1.0 and final 8.6+/-0.8 mg/24h). The effect of ramipril was abolished by HOE 140 (initial 11.6+/-2.0 and final 38.9+/ 11 mg/ 24 h). The systolic blood pressure of the controls increased from 106+/-2 to 144+/-5 mm Hg at the 14th week. Ramipril abolished the increase in systolic blood pressure. The effect of ramipril was reverted by HOE 140 (initial 108+/-2 and final 140+/-9 mmHg). Control rats had more severe histopathologic changes. Those animals receiving ramipril + HOE 140 displayed less severe glomerular changes, while rats treated only with ramipril had mild alterations. Thus the glomerular injury score was 2.11+/-0.32 for controls, 1.53+/-0.52 for rats treated with ramipril + HOE 140, and 0.06+/-0.04 for rats treated only with ramipril. The glomerular area was 20,886+/-1,410, 19,693+/-2,200 and 14,352+/ 3,200 microm2, respectively, for the 3 groups. These results suggest that the protective effect of ACEIs in the development of chronic renal failure is partially mediated by kinins. PMID- 9851321 TI - Nosocomial wound infections: a prevalence study and analysis of risk factors. AB - This first German prevalence study surveilling nosocomial wound infections (NWI) was carried out in 72 representatively selected hospitals. NWI were recorded by 4 validated investigators. Seventy-nine NWI were recorded among 4983 operated patients (prevalence rate: 1.61%), most of them after amputation of limbs (6.1%) and operations on the colon or rectum (3.1%). The 3 risk factors from the NNIS (National Nosocomial Infection Surveillance) index for postoperative wound infections were for the first time applied in this prevalence study. Patients with contaminated wounds had significantly more wound infections (P=0.01, likelihood ratio test) whereas, in contrast to the NNIS index, patients with ASA score >3 (P= 0.07) or long lasting operations (>75th percentile) did not (P=0.1). Therefore, for the stratification of NWI rates wound contamination is the single most important factor. PMID- 9851322 TI - Our experience with 2693 wounded treated at the Split University Hospital during the 1991-1995 period. AB - The authors' personal experience of 2693 wounded treated at the Split University Hospital during the 1991-1995 period is described and compared with the results reported from other recent wars worldwide. Explosive wounds were more frequent than gunshot wounds (N=1490; 55.3% vs N=988; 36.7%), and wounds due to other factors were observed in 215 (8.0%) patients. There were 2494 (92.6%) men and 99 (7.4%) women. A total of 1815 (67.4%) patients were operated on in field war hospitals, and 878 (32.6%) at the Split University Hospital. Recovery and discharge were achieved in 1527 (56.7%) and improvement with the treatment or rehabilitation continued at other institutions in 850 (31.6%) patients. In 240 (8.9%) patients, no definite opinion can yet be given. A lethal outcome was recorded in 76 (2.8%) patients. Rapid transportation from the site of wounding to medical teams was found to be of paramount importance for successful treatment. These teams were placed in field war hospitals placed as close as possible (5-15 km) to the frontline. PMID- 9851323 TI - Extended membranous tracheobronchial rupture after tracheal intubation. AB - Membranous tracheo-bronchial rupture due to tracheal intubation has been infrequently reported. We present a previous unreported case of an extended membranous tracheo-bronchial laceration successfully repaired, on an emergency basis, through a right thoracotomy and cervicotomy with primary multiple interrupted sutures without the use of a free patch. We believe that in this complication the prognosis is generally good and much more linked to the underlying disease than to tracheal rupture. PMID- 9851324 TI - A prospective study comparing vertical banded gastroplasty versus laparoscopic adjustable gastric banding in the treatment of morbid and super-obesity. AB - We present our prospective comparative study on 60 patients subjected to two types of gastric restrictive procedures, i.e. Vertical Banded Gastroplasty (VBG) and Laparoscopic Adjustable Gastric Banding (LAGB) at King Abdul Aziz University Hospital, Jeddah, Saudi Arabia. The aim of our study was to compare weight reduction after both procedures. Thirty patients had VBG and thirty had LAGB based upon informed consents. The age, sex and preoperative weight and BMI were comparable in both groups. Excess weight loss of 87% was achieved in the VBG group while only 50% was noticed in the LAGB group at 6 months postoperatively. PMID- 9851325 TI - Finsterer-Bancroft operation: an option for the treatment of difficult duodenal ulcers. AB - BACKGROUND: In the surgical management of duodenal ulcers, the most feared complications are related to the treatment of the duodenal stump after Billroth lI-type gastric resections. Such complications are more evident in so-called 'difficult duodenum' cases, whose identification is directly related to the surgeon's experience. Among available techniques to avoid those complications, one is the treatment of the antralduodenal stump by the method of pre-pyloric exclusion and removal of antral mucosa, as proposed by Finsterer in 1918 and diffused by Bancroft in 1932. This method, however, was criticized, especially because of the possibility of retaining residual antral mucosa, which would be a determinant factor for the ulcer disease recurrence. The objective of the study was to verify whether the Finsterer-Bancroft operation is a valid alternative in the treatment of unresectable duodenal ulcers, as well as to encourage its application by less experienced surgeons, by the standardization of the surgical technique. METHODS: From April, 1984 to December, 1996 two hundred and six elective partial gastrectomies for duodenal ulcers were performed with Billroth II reconstruction. Of these, in thirty-one (15%), the Finsterer-Bancroft method was used. The patients' ages, varied between 23 and 65 years, constituting 25 males and 6 females. In all cases, surgery was indicated due to the presence of stenosis. RESULTS: Three patients (9.7%) had complications. There was one death (3.2%) due to leakage of duodenal stump and peritonitis, one case of duodenal fistula (3.2%), and one case of ulcer recurrence (3.2%). All three complications were caused by inappropriate application of the method. CONCLUSIONS: We conclude that the Finsterer-Bancroft operation is a valid alternative in the surgical treatment of chronic duodenal ulcers, when considered unresectable, and is within the reach of in-training and less experienced surgeons. PMID- 9851326 TI - Surgical treatment for biliary lithiasis in patients associated with chronic renal failure on hemodialysis. AB - BACKGROUND: Incidences of the variety of postoperative complications in surgical treatment for patients with chronic renal failure on hemodialysis is the clinically accepted opinion. The aim of this study was to ascertain the safety of surgical treatment for patients of biliary lithiasis with chronic renal failure on hemodialysis. METHODS: Eleven consecutive patients with biliary lithiasis, who had a history of chronic renal failure treated with hemodialysis, were selected. The peri-operative course in-each of these cases was investigated. RESULTS: There was no incidence of peri-operative problems and postoperative complications in cases treated with laparoscopic cholecystectomy. One case treated with cholecystectomy with laparotomy had a slight degree of postoperative pneumonia, which soon improved after administration of antibiotics. Only one case, treated with choledochotomy, resulted in operative death due to accidental septic shock from an unknown origin. CONCLUSIONS: Surgical treatment of patients with biliary lithiasis and chronic renal failure on hemodialysis can be safely performed with appropriate operative indications and noninvasive surgical techniques. PMID- 9851327 TI - Surgical treatment for proximal bile duct carcinoma. AB - BACKGROUND: The surgical management of proximal bile duct carcinoma is controversial. There is no consensus among surgeons as to the indications for radical resection. This article described personal experience with the different surgical procedures for patients with proximal bile duct carcinoma. METHODS: The medical records of fifty-two consecutive patients undergoing surgical resection over a 20-year period were retrospectively analysed in terms of pathology, perioperative mortality, clinical course, and overall survival. RESULTS: The 1-, 3- and 5-year survivals for the entire group were 63.3%, 24.5%, and 21.0%, respectively. The results for local resection of the extrahepatic bile duct in 35 cases were unsatisfactory. Radical resection that included the right extended hepatic lobe, caudate lobe, and extrahepatic bile duct was performed in 9 patients and demonstrated an excellent 5 year survival rate of 44.4%. However, hospital mortality was 22.2%. There was no significant difference among the operative procedures for patients with advanced disease. CONCLUSION: Radical resection remains the procedure of choice in proximal bile duct carcinoma. However, results with surgical therapy alone remain unsatisfactory. Multimodality treatment that also includes radiotherapy and/or chemotherapy is recommended. PMID- 9851328 TI - The pattern of lymphatic spread in carcinoma of the distal bile duct. AB - From 1995 to 1996, 20 patients with carcinoma of the distal bile duct (Bi) and 16 with carcinoma of the pancreatic head (Ph) underwent pancreaticoduodenectomy or total pancreatectomy with systematic extended lymphadenectomy. Histopathological specimens were examined with special reference to nodal metastases. Pancreatic parenchymal invasion was present in 10 of 20 patients with Bi. The frequency of nodal involvement in Bi with pancreatic invasion and Ph was significantly greater than in Bi without pancreatic invasion (p<0.05 and p<0.01, respectively). Metastasis to nodes around the superior mesenteric artery or abdominal aorta in Bi with pancreatic invasion occurred more frequently than in Bi without pancreatic invasion (p<0.05), though was not different from the frequency observed in Ph. The pattern of lymphatic spread in distal bile duct carcinoma with pancreatic invasion was practically equal to the mode in carcinoma of the pancreatic head. PMID- 9851329 TI - Improved technique for pancreaticogastrostomy with implantation method after pancreaticoduodenectomy. AB - We performed improved pancreaticogastrostomy after pancreaticoduodenectomy in 60 consecutive patients between August, 1993 and July, 1997. Our improved pancreaticogastrostomy was made by implantation method with pancreatic duct stent. Two-layer fusion of anastomosis between full thickness or mucosa/submucosa of gastric wall and pancreatic parenchyma was completed via gastric lumen with gastrotomy in the anterior gastric wall. The morbidity and mortality rates were 3% and 0%, respectively. There was absolutely no pancreatic leakage, and no postoperative complications directly related to the pancreaticogastrostomy. The biggest advantage of this procedure was to be able to anastomose in a good field of vision via gastric lumen with gastrotomy in the anterior gastric wall. This improved pancreaticogastrostomy seemed to be technically very easy and a safe method. PMID- 9851330 TI - Delayed massive intraperitoneal hemorrhage after pancreatoduodenectomy. AB - Of 144 cases of pancreatoduodenectomy we treated 6 patients (4.2%) with delayed massive bleeding. Massive hemorrhage occurred 14 to 38 days later. Leakage of the pancreatojejunostomy was confirmed in all patients. A "sentinel bleed" was evident in five patients with arterial bleeding. One patient with hemorrhage of the superior mesenteric vein was successfully treated conservatively. One patient with bleeding from the left gastric artery stump survived emergency re-operation. Two of four patients with hemorrhage from the gastroduodenal artery stump were successfully treated with selective embolization of the common hepatic artery. The remaining two patients died of uncontrollable re-bleeding or hepatic failure following hemostasis. Angiography and selective embolization are effective for identification and control of the bleeding site when delayed hemorrhage occurs after pancreatoduodenectomy. Intensive treatment is necessary to compensate for reduced hepatic arterial blood supply in cases requiring surgical or radiological interruption of the common hepatic artery. PMID- 9851331 TI - Long-term morphological changes of remnant pancreas and biliary tree after pancreatoduodenectomy on CT. AB - BACKGROUND: Despite the increasing number of long-term survivors after pancreatoduodenectomy, little is known about morphological appearance of the remnant pancreas or the biliary system after surgery. METHODS: To evaluate long term morphological changes of the remnant pancreas and biliary tree after pancreatoduodenectomy, computed tomograms obtained at the time of surgery and more than 2 years after operation were reviewed in 19 Japanese patients. RESULTS: Two to three years after surgery, parenchymal atrophy of the remnant pancreas occurred in 9 (56%) of 16 patients without atrophy at the time of surgery. Three patients who had had parenchymal atrophy preoperatively showed no change after surgery. Of 10 patients with a dilated pancreatic duct preoperatively, 8 (80%) patients demonstrated a decline in ductal size, while the other 2 showed persistent ductal dilatation. Of 9 patients with a normal-sized pancreatic duct preoperatively, 2 patients (22%) developed ductal dilatation after surgery, and the other 7 showed no change. In 4 (57%) of 7 patients with a dilated hepatic duct preoperatively dilatation was reduced after surgery, whereas it persisted in the remaining 3. Only one (8%) of 12 patients with a nondilated biliary tree preoperatively showed ductal dilatation 6 months following surgery. Pneumobilia was revealed in 13 (68%) of 19 patients by the follow-up examinations. CONCLUSIONS: Surgeons should be aware of these changes in morphology of the remnant pancreas and biliary tree after pancreatectoduodenectomy. PMID- 9851332 TI - Evaluation of angiographically assisted spiral CT in surgical cases of hepatocellular carcinoma considering recurrent cases. AB - Between June 1993 and July 1996, computed tomography during arterial portgraphy (CTAP) was performed on 22 patients with hepatocellular carcinoma (HCC) and liver cirrhosis before surgery. In 17 out of 22 patients, CT hepatic arteriography (CTA) was combined with CTAP. All 22 patients underwent definitive surgery. In 16 patients in which CTA was successfully performed, 20 HCC nodules and 28 pseudolesions were detected. The pseudolesions were depicted larger in size with CTA than with CTAP, formed a variety of shapes, and were enhanced homogeneously, while HCC nodules were depicted larger with CTAP than with CTA, formed a round or wedge shape, and were sometimes enhanced heterogeneously. Recurrence was diagnosed in 9 of 22 patients. Five recurrent lesions in 4 patients were already revealed by pre-operative CTAP. CTA was performed in only 1 of 5 lesions. The diagnostic accuracy for HCC significantly improves by recognizing by recognizing pseudolesions with CTA and CTAP. PMID- 9851333 TI - Prognostic factors in resected hepatocellular carcinomas and therapeutic value of transcatheter arterial embolization for recurrences. AB - The authors summarize the results of patients who had hepatectomy for hepatocellular carcinoma (HCC) over a 16-year period. Survival rates for 52 patients with HCC classified according to the Liver Cancer Study Group of Japan were calculated according to various clinicopathological variables. A univariable analysis revealed that alpha-fetoprotein > or =2000 ng/ml, portal involvement, tumor size > or =3.1 cm, and noncurative resection were associated with unfavorable outcomes, while neither the number of tumors nor underlying cirrhosis was associated with such outcomes. Furthermore, some of the patients with recurrence survived long after transcatheter arterial embolization (TAE) for recurrent tumors. Early detection as well as TAE for recurrent HCCs is necessary to improve long-term survival. PMID- 9851334 TI - Decrease in plasma levels of vasoactive intestinal polypeptide in short bowel syndrome: experimental study. AB - To investigate the behaviour of the vasoactive intestinal polypeptide (VIP) in short bowel syndrome (SBS), an experimental model of massive intestinal resection (MIR) was developed. For this purpose, 20 'minipigs' were divided into two experimental groups: A (control) and B (MIR). The parameters determined were the mean plasma levels of VIP and the degree of steatorrhea at four different times: T1 (basal), T2 (one week after surgery), T3 (two weeks after surgery), and T4 (24 weeks after surgery). The results indicated that, after MIR, a progressive decrease in the mean plasma levels of VIP takes place, with statistical significance in T3 (p < 0.05) and T4 (p < 0.01). This situation seems to be a direct result of the massive loss of intestinal tissue, and could lead to the use of this peptide to mark the evolution of the intestinal adaptation process. PMID- 9851335 TI - Crohn's disease in infancy: a case report. AB - Crohn's disease in infants is infrequent. We describe here the case of a baby girl born at the 33rd week of pregnancy, that underwent surgery at the age of 55 days for intestinal occlusion. This was found to be due to stenosis of the right colonic flexure associated with cecum, appendix and terminal ileum. Microscopic examination showed features consistent with Crohn's disease. Eleven years after surgery, followed by cortisone therapy, the patient shows normal growth and is asymptomatic. PMID- 9851336 TI - The value of endoscopic ultrasonography in preoperative evaluation of rectal cancer. AB - BACKGROUND: In rectal cancer, preoperative evaluation of the depth of cancer invasion and the presence of pararectal lymph node metastasis is essential to determine appropriate operative procedures, especially in autonomic nerve preserving operations. For this evaluation, endoscopic ultrasonography (EUS) was performed prospectively in the following cases. METHODS: During the past 5 years, 70 patients with rectal cancer underwent EUS examination by using 7.5MHz and 12MHz-EUS sonoprobe systems. RESULTS: The depth of cancer invasion was accurately predicted by EUS in 76% of cases (53/70). Seventeen cases were incorrectly estimated, 12 of those underestimated, and 5 overestimated. The evaluation of pararectal lymph node metastasis was accurate in 44 patients (69%). The sensitivity rate was 36% and specificity was 90%. CONCLUSIONS: EUS is a very useful technique for determining therapeutic procedures for rectal cancer, especially in deciding whether or not sphincter preserving and/or autonomic nerve preserving operations should be undertaken. PMID- 9851337 TI - The prediction of the prognosis for rectal carcinoma down-staged by pre-operative radiotherapy by transrectal ultrasonography. AB - BACKGROUND: We attempted to determine whether the results of transrectal ultrasonography (TRUS) could provide any prognostic information about patients with rectal carcinoma treated with pre-operative radiotherapy (RT). METHODS: Ninety-eight patients with rectal carcinoma were treated with pre-operative RT and radical surgery. We categorized the TRUS results with respect to TNM classification before the RT (Pre-RT staging), those after the RT (Post-RT staging), and the results of the pathological examination (Post-OP staging). We analysed the associations among these three stagings, and the survival time of the patients. RESULTS: The prevalence of under T2-stage (TNM classification) in the pre-RT staging was significantly lower than that in both the Post-RT and Post OP staging. The number of tumours diagnosed as T4 stage in both the Post-RT and Post-OP staging was significantly lower than that before irradiation. The survival periods of the patients with tumours remaining as T4 after radiation were significantly shorter than those of the patients diagnosed as having T4 tumours at the Pre-RT staging (Pre-RT versus Post-RT, p = 0.019; Pre-RT versus Post-OP, p = 0.030). These results suggested that radio-insensitive residual cells might associate with the poor prognosis. CONCLUSION: We could predict the possible prognosis in rectal carcinoma patients treated with pre-operative radiotherapy before operation by TRUS. PMID- 9851338 TI - Internal hernia in adults. AB - BACKGROUND: Congenital bands, congenital anomalies of the intestinal rotation and congenital defects in the mesentery or omentum, postsurgical or traumatic defects of the mesentery and omentum are important factors that predispose to internal herniation. In this retrospective study, 18 cases of internal herniation were evaluated and the factors that effect the prognosis were investigated. METHODS: The records of 609 patients who underwent laparotomy due to acute intestinal obstruction were retrospectively reviewed. The cause of intestinal obstruction was internal herniation in 18 patients. The patients were evaluated with respect to signs and symptoms, radiological findings, time elapsed between the onset of symptoms and surgery, findings, in surgery, type of operation performed, postoperative complications and postoperative stay. RESULTS: Postoperative complications were encountered more frequently in the patients with bowel strangulation (p=0.045) in our series. Time elapsed between the onset of symptoms and laparotomy was found to be significantly longer in the patients with strangulation than in those without strangulation (p=0.046). Additionally, postoperative hospital stay was shorter in the patients without strangulation (p=0.03). CONCLUSIONS: Since preoperative diagnosis of an internal herniation is very difficult because of the lack of specific signs and symptoms, postoperative complications can only be decreased with early surgical intervention in the patients with acute intestinal obstruction. PMID- 9851339 TI - Umbilical hernial defects encountered before and after abdominal laparoscopic procedures. AB - Pre-existing fascial umbilical defects may compound problems faced during laparoscopic surgery. Detection helps in preventing bowel or omental injury and repair can be done at the same time. Postoperative port incisional hernias are not uncommon and require surgical repair. METHODS: The incidence, clinical features and management of such fascial defects were studied in 2100 patients undergoing abdominal laparoscopy. RESULTS: The incidence of fascial defects was 18%. The hernias were symptomatic in 56.5% cases, with an overwhelming female preponderance. A supra-umbilical incision above the upper limit of the hernia was used to establish the umbilical port and through this the hernias were repaired with nonabsorbable sutures. Postoperative incisional hernias occurred in 41 patients out of a total follow-up of 1892 cases (2.16%). The common predisposing factors were wound infection, postoperative chest infection and previous existing diseases, like diabetes and connective tissue disorders. Twenty-eight patients underwent mesh plasty and 3 cases had laparoscopic Gortex mesh repair. The remaining 10 cases refused surgery. CONCLUSIONS: Omental fascial defects should be dealt with at the same sitting through a supra-umbilical port incision avoiding direct entry into the hernia. Use of nonabsorbable sutures is recommended. Prevention of wound infection and postoperative chest infection greatly reduces the chances of an incisional hernia. PMID- 9851340 TI - Laparoscopic versus open preperitoneal prosthetic herniorrhaphy for recurrent inguinal hernia. AB - BACKGROUND: Laparoscopic herniorrhaphy (LH) shares the same repair principle as open preperitoneal prosthetic herniorrhaphy (PPH). Theoretically, the recurrence rate of LH for recurrent inguinal hernia will match the low recurrence rate of PPH (1.2-3%). METHODS: One-hundred forty-five cases of recurrent inguinal hernia were retrospectively studied between 1990 and 1994. Forty-two cases receiving LH were compared to 103 cases receiving PPH. RESULTS: There were no differences in operative time, hospital stay, morbidity rate, satisfaction scale and recurrence rate between the LH group and the PPH group. The LH group showed significantly less postoperative pain and 2 times shorter convalescence (p<0.01). Unsuspected asymptomatic contralateral hernia was found in 4.8% of patients receiving LH. 11.9% of patients had bilateral hernia repairing at the same time in the LH group. CONCLUSIONS: LH is suitable for recurrent inguinal hernia, but further investigation of this technique is required before its wide application. PMID- 9851341 TI - Use of vascular closure staples in vascular access for dialysis, kidney and pancreas transplantation. AB - We report our experience with the use of the vascular closure staples (VCS) in vascular access for dialysis, as well as in kidney and pancreas transplantation. We used the VCS for 50 endogenous arterio-venous fistulas (AVFs). There were no complications. The use of the VCS contributed in creating an excellent anastomosis and minimising operative time. All AVFs are in use for dialysis (follow up two months to one year). The excellent results from our experience with the use of VCS for vascular access encouraged us to use them in kidney and pancreas transplantation. We performed six cadaveric kidney transplants (the first operation was the first application of the VCS in kidney transplantation in Europe) and two cadaveric pancreas-kidney transplants (the first operation being the first application of the VCS in pancreas transplantation in the world). There were no complications. The use of VCS created an excellent anastomosis and minimised warm ischaemia time. All kidney transplant recipients have normal creatinines (follow up 1-5 months) and the recipients of pancreatic transplants are insulin independent (follow up 1-3 months). PMID- 9851342 TI - Incidental cholecystectomy in the elderly. PMID- 9851343 TI - Total thyroidectomy for multinodular goiter. PMID- 9851344 TI - Moving moments in orthodontics. PMID- 9851345 TI - Soft tissue evaluation of contemporary Caucasian and African American female facial profiles. AB - Previous studies suggest that esthetic Caucasian profiles exhibit fuller lips than the norm for their race, while esthetic African American profiles are similar to those of esthetic Caucasians. The present study was undertaken to compare the profiles of female Caucasian and African American models and their nonmodel counterparts. Four groups of 30 subjects were evaluated: Caucasian models [CM], Caucasian controls [CC], African American models [AM], and African American controls [AC]. The models' profiles were photographed from current fashion magazines, the photos were scanned, and 17 landmarks were digitized. Each profile was standardized for size and oriented along the N'-Sn' line on a Macintosh 6115CD computer. Control photographs were processed in a similar manner. Twenty-six variables were measured for each profile. Means, ranges, and standard deviations were computed along with unpaired, two-tailed Student's t tests (p<0.05) to evaluate group differences. The results showed that for the AM and AC profiles, all but two of the 26 variable were similar. For the CM and CC profiles, eight variables demonstrated significant differences. Between-race comparisons demonstrated greater numbers of parameters that were significantly different: CM/AM with 18 and CM/AC, CC/AC, and CC/AM with 22 each. Most of the differences involved the lips. Vertical soft tissue proportions for the four groups did not follow a 40/20/40 ratio. Caucasian and African American models displayed significantly different profile characteristics. The African American models and controls showed similar profile features, whereas greater differences were observed between Caucasian models and controls. Based on our study, the African American profile currently presented in the mass media is not "Caucasian like." In fact, it appears that Caucasian models display more ethnic features than African American models do Caucasian features, suggesting that previously held concepts of facial beauty may no longer apply. PMID- 9851346 TI - Cephalometric floating norms for North American adults. AB - Floating norms provide a method of analysis that uses the variability of the associations among suitable cephalometric measures, on the basis of a regression model combining both sagittal and vertical skeletal parameters. This study establishes floating norms for the description of the individual skeletal pattern in North American adults. The method is based on the correlations among the following craniofacial measurements: SNA, SNB, NL-NSL, ML-NSL, and NSBa. The results are given in a graphical box-like form. This easy, practical procedure allows for the identification of either individual harmonious craniofacial features or anomalous deviations from the individual norm. The use of cephalometric floating norms may be helpful for diagnosis and treatment planning in orthognathic surgery and dentofacial orthopedics. PMID- 9851347 TI - Determining cephalometric norms for Caucasians and African Americans in Birmingham. AB - The purpose of this study was to establish age- and sex-specific normative data for Caucasians and African Americans in Birmingham, Ala. Subjects (136) between 6 and 18 years old were included in the study. Chang's method (AF-BF) and 12 other measurements were used as determinates of the skeletal sagittal jaw relationship. All subjects had acceptable facial profiles and Class I occlusion. Subjects were divided into eight subgroups based on race, gender, and age. Differences of mean cephalometric values were tested using parametric and nonparametric statistical tests. Compared with Caucasians, African Americans had greater mean values for all measurements except AFB and AF-BF. More negative values were found for the African Americans in the Wits appraisal. Most measurements were found to decrease with age. These findings support our hypothesis that cephalometric norms should be based on racial, sex, and age differences. PMID- 9851348 TI - Consistency of patient classification in orthodontic diagnosis and treatment planning. AB - The purpose of this study was to examine diagnosis and treatment planning categorizations used by experienced orthodontists. Thirty-nine clinicians were asked to evaluate six test cases and formulate a diagnosis and treatment plan for each. The information provided was categorized using a problem-oriented classification. The results indicate little agreement exists in diagnostic subcategories, such as molar relationship, degree of crowding, or the nature of skeletal discrepancies. There was also little agreement regarding some treatment strategies, such as extraction of teeth, the use of orthopedic appliances, or the use of surgery. A need exists for better definitions of diagnostic criteria and appropriate treatment options. PMID- 9851349 TI - A retrospective study of two-stage treatment outcomes assessed with two modified PAR indices. AB - This retrospective study was undertaken to evaluate the long-term outcome of two stage (functional/fixed) Class II treatment. A modified peer assessment rating (PAR) was applied to the records of 27 patients who had been recalled an average of 9 years after the completion of the second phase of treatment. UK and US weightings were applied. Analysis of variance identified significant differences among treatment stages. The greatest change in PAR score occurred during the first (functional) phase of treatment. By the end of the second phase, there had been an 83% reduction in PAR score. At recall, however, the PAR scores had increased significantly, due largely to relapse in overjet and in the lower labial segment. These results call into question the ultimate utility of early, two-stage treatment regimens. Although the differences between the UK and USA weightings were smaller than anticipated, the nature of the relapse seen here argues against the American exclusion of the lower labial segment. PMID- 9851350 TI - An assessment of extraction versus nonextraction orthodontic treatment using the peer assessment rating (PAR) index. AB - The extraction of teeth for orthodontic purposes has always been a controversial subject in the specialty. The purpose of this study was to assess the outcome of orthodontic treatment in 100 patients treated with the extraction of four premolars and compare it with the outcome of 100 patients treated without extractions, using the peer assessment rating (PAR) index. Records were selected from 1,198 consecutively completed cases treated by a single provider (MGH) between 1981 and 1995. We chose the first 100 finished patients in each group (extraction and nonextraction) who were under the age of 16 and had no deciduous teeth at the start of treatment. The results of this study show that average treatment time for the extraction group was 29.7+/-6.1 months compared with 26.0+/-7.2 months for the nonextraction group. The extraction group had significantly higher initial PAR scores (T1-PAR ext = 30.01+/-8.20 vs. T1 PAR nonext = 25.21+/-8.55), with greater initial maxillary anterior crowding (PAR ext value = 6.05+/-3.85 vs. PAR nonext value = 4.21+/-2.90) and greater initial overjet (PAR ext value = 1.82+/-1.01 vs. PAR nonext value 1.28+/-1.04). All pretreatment differences were significant at the p < or = 0.0001 level. Although significantly different at the beginning of treatment, both groups were statistically identical at the end (PAR T2 ext = 6.18 + 3.04% reduction = 79.4% compared with PAR T2 nonext = 5.64 + 3.08% reduction = 77.6%). In conclusion, the results demonstrate that, given an additional 3 months of treatment, it is possible for an orthodontist to produce dento-occlusal relationships in extraction patients that are as good as those achieved in nonextraction cases. PMID- 9851351 TI - Earlier dental maturation: fact or fiction? AB - The objective of this investigation was to determine if there is a difference in dental age of maturation between adolescents treated in the 1970s and those treated in the 1990s. Records of 150 Caucasian patients, 8.5 to 14.5 years old and treated in a private orthodontic office between 1972 and 1974, were randomly selected; records of another 150 patients of the same race and age range but treated between 1992 and 1994 were also collected. The percentage of calcification of the mandibular canines was rated according to methods used by Demirjian, who divided tooth development into eight segments, A to H. Using stage G to compare the 1970 and 1990 patient samples, we demonstrated dental age reductions of 1.21 years for males and 1.52 years for females, and a combined reduction of 1.40 years. PMID- 9851352 TI - Facial changes in extraction and nonextraction patients. AB - The purpose of this study was to determine if there are any differences in the posttreatment faces of patients treated nonextraction and those treated with premolar extraction in properly diagnosed and treated cases. The study was conducted in two parts. In Part 1, perceptions of 192 experienced general dentists and orthodontists were tested. Facial photos of 25 consecutively treated nonextraction patients and 25 consecutively treated four-premolar-extraction patients were shown to the study participants. They were asked if the patient was treated with the extraction of four premolars or without extractions. The mean score of the respondents was 54%, only slightly better than pure chance. In Part 2, profiles were evaluated based on cephalometric tracings. There was no significant difference between pretreatment and posttreatment profiles of the groups. The mean H-line values for both groups were within the desired esthetic range. It was concluded that experienced orthodontists and general dentists could not determine whether treatment was nonextraction or extraction by looking at the face alone. Also, there was no significant difference between the faces produced by the two types of treatments. Therefore, the avoidance of extracting premolars based on a fear of a significant detrimental effect on the face is unjustified when the case has been properly diagnosed and treated. PMID- 9851353 TI - Dentofacial disharmony: psychological status of patients seeking treatment consultation. AB - A sample of 194 patients whose dentofacial disharmonies were severe enough to warrant an orthognathic surgical treatment option completed the SCL-90-R, a 90 item assessment tool used to measure current level of psychological distress. Two global and nine primary dimension scores of psychological distress were computed. The average interpersonal sensitivity, psychoticism, and obsessive-compulsive dimension scores were higher than the nonpsychiatric patient population norms for both males and females, but the confidence intervals for these dimensions were still in the upper end of the normal range of functioning (< 1 SD above the normative mean). No statistically significant differences by gender or age group were observed. Over 15% of the patients were clinically elevated on obsessive compulsive behavior, interpersonal sensitivity problems, hostility, paranoid ideation, and psychoticism; and 24.7% qualified as a positive diagnosis for a psychiatric disorder. It appears that a surprisingly large number of individuals with dentofacial disharmonies who are seeking treatment consultation are experiencing a level of psychological distress that warrants intervention. PMID- 9851354 TI - Relationship between facial types and tooth and bone characteristics of the mandible obtained by CT scanning. AB - The purpose of this study was to evaluate relationships between morphological characteristics of vertical sections of the mandibular body and facial type. Of the correlation coefficients between tooth and bone inclination and facial type parameters, facial height index (FHI) was negatively associated with second premolar (P2), first molar (M1), and second molar (M2) inclinations. The angle represented by Frankfort horizontal plane to mandibular plane (FMA) was negatively associated with bone inclination of the M2 section. The buccal cortical bone was thicker in short-faced individuals than in the average and long faced groups, while lingual cortical bone thickness of the M1 and M2 sections was greater. The basal cortical bone thickness of the L1 section was greater in the short-faced group, and the inclinations of the P2, M1, and M2 axes were significantly smaller. Teeth in the short-faced group inclined more lingually than in the average- and long-faced groups. The results of this study provide evidence that a significant but complex relationship exists between structures of the mandibular body and facial types. The morphological features that relate to masticatory function and facial types are associated with cortical bone thickness of the mandibular body and the buccolingual inclination of the first and second molars. PMID- 9851355 TI - Maturation of B cells in the lamina propria of human gut and bronchi in the first months of human life. AB - Little is known of the maturation of the mucosae-associated lymphoid tissue (MALT) in man, because, for ethical reasons, tissues from newborns are not easy to obtain. We used the opportunity provided by autopsies systematically performed in infants who died of Sudden Infant Death Syndrome (SIDS) to study the maturation of the MALT after birth. Gut and bronchus samples of 90 infants from postpartum to 90 months and who died from SIDS were collected and studied by histological and immunofluorescence examination. Plasma cells, absent at birth, appeared within a few hours after birth and initially were of the IgM isotype. IgA plasma cells appeared at 12 days. These cells were first observed in gut and later in bronchi, indicating that maturation of the gut precedes that of bronchi. The number of plasma cells increased rapidly over time and IgA plasma cells became predominant after 3 weeks in the gut and 6 weeks in bronchi. At birth, only small IgM bearing B-cell foci were seen and organized germinal centers appeared to develop over a few days, first in the gut and only later in bronchi. These results confirm that, in man, the MALT organization at birth is still in its fetal form and that maturation depends on intestinal challenges and evolves over several weeks before IgA becomes the predominant isotype secreted. PMID- 9851356 TI - Positive and negative selection in the thymus and the thymic paradox. PMID- 9851357 TI - Epidermal growth factor modulates fetal thymocyte growth and differentiation. AB - In the present study, we used the fetal organ culture (FTOC) technique in order to study a putative effect of epidermal growth factor (EGF) on the thymus ontogeny. Functional EGF receptors and more recently the EGF molecule itself, respectively, on the membrane of epithelial components of thymic stroma and on a few thymocytes in adult thymus, had been reported in the literature. We could observe a dose-dependent decrease in cellularity and a progressive retention of thymocytes in the double-negative (CD4-/CD8-) stage of differentiation when exogenous EGF was added. Epidermal growth factor interfered with both fetal stroma growth and thymocyte development at a precise moment, that is, in the passage from double-negative to the double-positive (CD4+/CD8+) stage. After a 7 day FTOC in the presence of EGF, most cells recovered were Thy-1.2+, c-kit+, TSA1 /int, CD3-, and one of CD44high/CD25int, CD44-/CD25int, or CD44/CD25-. Some developed into gammadeltaTCR+ cells with a mature (CD3+) phenotype, but not into alphabetaTCR+ thymocytes. It seems that EGF addition makes the cultures "nonpermissible" for alphabetaTCR+ thymocyte generation. We report here the presence of a high Mr "EGF-like" molecule on the membrane of fetal thymocytes, which role in the observed effects is under investigation. Further biochemical characterization of this molecule is still required, because its presence was only evidenced on the basis of its antigenicity. PMID- 9851358 TI - Role of prolactin in the recovered T-cell development of early partially decapitated chicken embryo. AB - Although different experimental approaches have suggested certain regulation of the mammalian immune system by the neuroendocrine system, the precise factors involved in the process are largely unknown. In previous reports, we demonstrated important changes in the thymic development of chickens deprived of the major neuroendocrine centers by the removal of embryonic prosencephalon at 33-38 hr of incubation (DCx embryos) (Herradon et al., 1991; Moreno et al., 1995). In these embryos, there was a stopping of T-cell maturation that resulted in an accumulation of the most immature T-cell subsets (CD4-CD8- cells and CD4-CD8lo cells) and, accordingly, in decreased numbers of DP (CD4+CD8+) thymocytes and mature CD3+TcRalphabeta+ cells, but not CD3+TcRgammadelta lymphocytes. In the present work, we restore the thymic histology as well as the percentage of distinct T-cell subsets of DCx embryos by supplying recombinant chicken prolactin, grafting of embryonic pituitary gland, or making cephalic chick-quail chimeras. The recovery was not, however, whole and the percentage of CD3+TcRalphabeta+ thymocytes did not reach the normal values observed in 17-day old control Sham-DCx embryos. The results are discussed on the basis of a key role for prolactin in chicken T-cell maturation. This hormone could regulate the transition of DN (CD4 CD8 ) thymocytes to the DP (CD4+CD8+) cell compartment through its capacity for inducing IL-2 receptor expression on the former. PMID- 9851359 TI - Protective cellular immunity against influenza virus induced by plasmid inoculation of newborn mice. AB - Neonate organisms display an intrinsic disability to mount effective immune responses to infectious agents or conventional vaccines. Whereas low doses of antigens trigger a suboptimal response, higher doses are frequently associated with tolerance induction. We investigated the ability of a plasmid-expressing nucleoprotein of influenza virus to prime a specific cellular immune response when administered to newborn mice. We found that persistent exposure to antigen following plasmid inoculation of neonates leads to a vigorous priming of specific CTLs rather than tolerance induction. The CTLs were cross-reactive against multiple strains of type A influenza viruses and produced IFNgamma but no IL-4. The immunity triggered by plasmid inoculation of neonates was protective in terms of pulmonary virus clearance as well as survival rate following lethal challenge with influenza virus. Whereas the persistence of the plasmid at the site of injection was readily demonstrable in adult mice at 3 months after inoculation, mice immunized as newborns displayed no plasmid at 3 months and very little at 1 month after injection. Thus, DNA-based immunization of neonates may prove an effective and safe vaccination strategy for induction of cellular immunity against microbes that cause serious infectious diseases in the early period of life. PMID- 9851360 TI - Sequence of the rag1 and rag2 intergenic region in zebrafish (Danio rerio). PMID- 9851361 TI - Sequence analysis of the mouse RAG locus intergenic region. AB - The recombination activating genes RAG-1 and RAG-2 are highly conserved throughout evolution and are necessary and essential for the DNA rearrangement of antigen-receptor gene segments. These convergently transcribed genes are expressed primarily by developing B and T lineage cells. In addition, recent data suggest that the RAG locus can be reactivated in mouse germinal center B cells. Despite these well-defined patterns of expression, little is known about mechanism(s) regulating transcription of the RAG locus. Experiments with a mouse fibroblast line stably transfected with a genomic fragment of the RAG locus suggest that the intergenic region between RAG-1 and RAG-2 may contain information modulating RAG transcription. In order to begin testing this hypothesis, we have sequenced the 7.0-kb RAG intergenic region of the mouse. The sequence did not contain open reading frames larger than 60 amino acids. Analysis with GCG software identified several potential transcription-factor binding sequences within this region. Many of these are associated with transcriptional regulation of the Ig locus. PMID- 9851362 TI - Relation of inducible nitric oxide synthase activity to lipid peroxidation and nonprotein sulfhydryl oxidation in the development of stress-induced gastric mucosal lesions in rats. AB - We have reported that increases in lipid peroxide (LPO) formation, nonprotein sulfhydryl (NP-SH) oxidation, and inducible NO synthase (iNOS) activity and a decrease in constitutive NO synthase (cNOS) activity in the gastric mucosa of rats with water immersion restraint (WIR) stress are closely related to gastric mucosal lesion development. Peroxynitrite, which is produced by the reaction of nitric oxide (NO) with superoxide anion, can initiate intracellular LPO formation and NP-SH oxidation, resulting in producing an extreme cellular membrane damage. In this study, the relation of changes in cNOS and iNOS activities to LPO formation and NP-SH oxidation was examined in the gastric mucosa of rats with WIR stress. An increase in iNOS activity, but not a decrease in cNOS activity, correlated well with an increase in LPO concentration (r = 0.750) and NP-SH concentration (r = -0.808) in the gastric mucosa of rats with WIR stress. In addition, the above-mentioned changes in iNOS activity and LPO and NP-SH concentrations with lesion development in the gastric mucosa of rats with WIR stress were attenuated with both prevention of the lesion development and an increase in the concentration of gastric mucosal nitrite/nitrate, the breakdown products of NO, by pretreatment with aminoguanidine, a selective iNOS inhibitor. These results suggest that in the gastric mucosa of WIR-stressed rats, NO produced by increased iNOS could contribute to enhanced LPO formation and NP-SH oxidation, resulting in lesion development. PMID- 9851363 TI - The 2.03 signal as an indicator of dinitrosyl-iron complexes with thiol containing ligands. AB - The parameters of EPR signal from dinitrosyl-iron complexes (DNIC) with bovine serum albumin (BSA), horse hemoglobin (Hb), and apometallothionein (apo-Mt) of horse kidney incorporating one (BSA, Hb) or two thiol-containing ligands (apo-Mt) were compared. The EPR signal from DNIC-BSA was characterized by the rhombic symmetry of g tensor at room temperature of signal recording (ambient temperature) or at 77K in the solution frozen in the presence of glycerol. In freezing of the solution in the absence of glycerin, under the exposure of DNIC BSA to negatively charged sodium dodecyl sulfate (SDS) ions, or in the incorporation of DNIC-BSA into the reversed micelles formed by negatively charged ions of surfactant aerosol OT, the symmetry of the g tensor of DNIC-BSA EPR signal increased to axial. A similarly high symmetry of g tensor was observed for the DNIC-Hb EPR signal in the absence of any influence on this protein complex. The shape of EPR signals from these preparations recorded at 77K was identical to that of EPR signal from DNIC with cysteine in frozen solution. In this connection it was concluded that the EPR signal from this low-molecular DNIC with the (RS )2Fe+(NO+)2 structure cannot be considered as a peculiar "fingerprint" of DNIC with the same structure in biosystems. In such systems the same signal can originate from protein DNIC incorporating only one thiol-containing ligand along with a nonthiol ligand. The EPR signal displayed by DNIC with apo-Mt with a high content of cysteine residues at room temperature of registration was identical to the EPR signal from frozen solution of DNIC with cysteine. This protein DNIC is apparently characterized by the same structure as DNIC with cysteine. PMID- 9851364 TI - Ascorbate-dependent enhancement of nitric oxide formation in activated macrophages. AB - Macrophages activated with bacterial lipopolysaccharide (LPS) and cytokines produce nitric oxide through the induction of iNOS gene expression. Ascorbate increased NOx (nitrite and nitrate) formation by approximately 40% in a mouse macrophage-like cell line, J774.1, activated with LPS and interferon-gamma. Ascorbate alone exhibited no inductive activity toward NO formation. N(G) Monomethyl-L-arginine inhibited nitrite formation in cells activated in the presence or absence of ascorbate. Northern and Western blotting analyses showed that both iNOS mRNA and protein steady-state levels were increased approximately twofold in cells activated in the presence of ascorbate compared to in cells activated only with the inducers. These data suggest that ascorbate increased NO production by increasing the amount of iNOS in the activated macrophages. PMID- 9851365 TI - Complete coding sequence of inducible nitric oxide synthase from human heart and skeletal muscle of patients with chronic heart failure. AB - The presence of the inducible nitric oxide synthase (iNOS) has previously been reported in heart and skeletal muscle of patients with chronic heart failure, where it is thought to be involved in the pathogenesis of dilated cardiomyopathy or of exercise intolerance observed in patients with chronic heart failure. To identify the iNOS of these human tissues the cDNA sequence was determined using reverse transcription polymerase chain reaction (RT-PCR). The deduced amino acid sequence of human heart and skeletal muscle shows a 99% identity between each other. The amino acid sequence was distinct from the constitutively expressed nitric oxide synthases. A 98% identity at the protein level was detected to rat vascular smooth muscle iNOS, whereas the homology to human hepatocyte iNOS was only 79%. The cofactor binding sites in the reported iNOS from human heart and skeletal muscle are highly conserved. The iNOS sequence described in this report is the first one isolated directly from human tissue expressing iNOS in the circumstances of chronic heart failure. PMID- 9851367 TI - New aspects in the reaction mechanism of phenol with peroxynitrite: the role of phenoxy radicals. AB - The decomposition of peroxynitrite (PON) in aqueous solutions was investigated by monitoring the release of dioxygen as a function of pH together with the various reaction products generated from phenol. This substrate was used as a mechanistic model for tyrosine nitration in prostacyclin synthase for which we have reported a highly efficient nitration and inhibition by PON (Zou, M., Martin, C., and Ullrich, V. (1997) Biol Chem. 378, 707-713). Nitrite as a contaminant and product of PON generated 4-nitrosophenol and some nitrophenols in the acidic pH range. In the alkaline range high amounts of 4-nitrosophenol originated from the disproportionation of PON yielding dioxygen and NOx species. The hydroxylation of phenol occurred between pH 3 and 8 with a maximum at 4.5. The nitration by PON also required a pH between 4 and 8 but had a second maximum between 10 and 12, suggesting that in this pH range phenolate was the reacting species. All isomeric biphenols were found as dimerization products as well as 4-phenoxyphenol (4 hydroxydiphenyl ether), indicating phenoxy radicals as intermediates. Since anisol when incubated under the same conditions yielded only hydroxylation but virtually no nitration products, it was concluded that nitration of phenolic compounds requires a one-electron oxidation as a primary step, followed by addition of the nitrogen dioxide radical. PMID- 9851366 TI - Protein kinase C-independent selective induction of nitric oxide synthase activity in rat alveolar macrophages by staurosporine. AB - The purpose of this study was to characterize the effect of the K-252a family of protein kinase inhibitors with emphasis on staurosporine (ST), on stimulation of the inducible nitric oxide synthase activity in rat alveolar NR8383 macrophages. We found that ST, but not K-252a, K-252b, KT-5720, and KT-5823, selectively enhanced the basal or the lipopolysaccharide (LPS)-induced nitric oxide production. ST-induced NO production was blocked by L-NAME, K-252a, and phosphatase inhibitors and could not be mimicked by other protein kinase C (PKC) inhibitors such as calphostine. An additive effect between ST and PMA on NO production was observed. LPS and PMA but not ST induced PKCbeta translocation from the cytosol to the membrane fraction. ST may induce and affect the state of phosphorylation of iNOS via PKC-independent mechanisms. ST provides an important pharmacological tool to investigate PKC-independent signal transduction pathways which regulate iNOS, induction, and activity in rat NR8383 macrophages. PMID- 9851368 TI - Nitrogen oxides and hydroxyguanidines: formation of donors of nitric and nitrous oxides and possible relevance to nitrous oxide formation by nitric oxide synthase. AB - The involvement of nitric oxide in numerous biological functions has led to the intense study of nitric oxide (NO) generation by the nitric oxide synthases (NOS) responsible. In addition to NO, nitric oxide synthases produce N(G)-hydroxy-L arginine, superoxide anion and, indirectly, NOx species such as peroxynitrite and, possibly, nitrous oxide (N2O). Consequently, the interactions of N(G) hydroxy-L-arginine with NO and other oxides of nitrogen (NOx) are of considerable interest. N(G)-Hydroxy-L-arginine and other monosubstituted hydroxyguanidines react with aqueous aerobic NO, peroxynitrite, and various NOx and nitrosating agents to form compounds that subsequently release NO and N2O. Spectrometric data indicate that the nitrosation product of N(G)-hydroxy-L-arginine is of the same N nitroso-N-hydroxy/diazeniumdiolate (formerly "NONOate") structure as previously found for the nitrosation products of other model hydroxyguanidines. These decompose in aqueous solution in a pH-dependent manner to yield mainly NO and ureas at low pH, N2O and cyanamides at basic pH, and what appear to be primary nitrosamines/ nitrosoimines. Studies on purified iNOS using a mass spectrometer with a gas-permeable membrane inlet identified both NO and N2O (or 15NO and 15N15NO with 15N-labeled L-arginine as substrate) as products of NOS activity. These experiments suggest that much more NO than N2O is produced under the conditions studied and that N2O formation can be rationalized via the reaction of NOx species with N(G)-hydroxy-L-arginine. PMID- 9851369 TI - Microscopic observations of the different morphological changes caused by anti bacterial peptides on Klebsiella pneumoniae and HL-60 leukemia cells. AB - Natural anti-bacterial peptides cecropin B (CB) and its analogs cecropin B-1 (CB 1), cecropin B-2 (CB-2) and cecropin B-3 (CB-3) were prepared. The different characteristics of these peptides, with amphipathic/hydrophobic alpha-helices for CB, amphipathic/amphipathic alpha-helices for CB-1/CB-2, and hydrophobic/hydrophobic alpha-helices for CB-3, were used to study the morphological changes in the bacterial cell, Klebsiella pneumoniae and the leukemia cancer cell, HL-60, by scanning and transmission electron microscopies. The natural and analog peptides have comparable secondary structures as shown by circular dichroism measurements. This indicates that the potency of the peptides on cell membranes is dependent of the helical characteristics rather than the helical strength. The microscopic results show that the morphological changes of the cells treated with CB are distinguishably different from those treated with CB-1/CB-2, which are designed to have enhanced anti-cancer properties by having an extra amphipathic alpha-helix. The morphological differences may be due to their different modes of action on the cell membranes resulting in the different potencies with lower lethal concentration and higher concentration of 50% inhibition (IC50) of CB on bacterium and cancer cell, respectively, as compared with CB-1/:CB-2 (Chen et al. 1997. Biochim. Biophys. Acta 1336, 171-179). In contrast, CB-3 has little effect on either the bacterium or the cancer cell. These results provide microscopic evidence that different killing pathways are involved with the peptides. PMID- 9851370 TI - Solution structure of peptides from HIV-1 Vpr protein that cause membrane permeabilization and growth arrest. AB - Vpr, one of the accessory gene products encoded by HIV-1, is a 96-residue protein with a number of functions, including targeting of the viral pre-integration complex to the nucleus and inducing growth arrest of dividing cells. We have characterized by 2D NMR the solution conformations of bioactive synthetic peptide fragments of Vpr encompassing a pair of H(F/S)RIG sequence motifs (residues 71-75 and 78-82 of HIV-1 Vpr) that cause cell membrane permeabilization and death in yeast and mammalian cells. Due to limited solubility of the peptides in water, their structures were studied in aqueous trifluoroethanol. Peptide Vpr59-86 (residues 59-86 of Vpr) formed an alpha-helix encompassing residues 60-77, with a kink in the vicinity of residue 62. The first of the repeated sequence motifs (HFRIG) participated in the well-defined alpha-helical domain whereas the second (HSRIG) lay outside the helical domain and formed a reverse turn followed by a less ordered region. On the other hand, peptides Vpr71-82 and Vpr71-96, in which the sequence motifs were located at the N-terminus, were largely unstructured under similar conditions, as judged by their C(alpha)H chemical shifts. Thus, the HFRIG and HSRIG motifs adopt alpha-helical and turn structures, respectively, when preceded by a helical structure, but are largely unstructured in isolation. The implications of these findings for interpretation of the structure-function relationships of synthetic peptides containing these motifs are discussed. PMID- 9851371 TI - Biological and conformational studies on analogues of a synthetic peptide enhancing HIV-1 infection. AB - We have previously demonstrated that a 23-amino acid peptide derived from the V3 loop of the surface glycoprotein of the HIV-1 strain MN is able to bind CD4 and to enhance HIV-1 infection. Further studies have suggested that the peptide/CD4 interaction induces an increase in both CD4 expression and CD4/gp120 binding affinity. This paper describes the biological and physico-chemical characterization of three analogues of reduced sequence that have been designed in order to identify the minimum active sequence of this peptide corresponding to the MN-HIV- 1 principal neutralizing domain. Biological studies indicate that the entire sequence is required for biological activity and that the sequence 1-18 presents an inhibitory activity. CD and FT-IR absorption data are discussed here in order to identify possible structure-function correlations. PMID- 9851372 TI - Tuning micelles of a bioactive heptapeptide biosurfactant via extrinsically induced conformational transition of surfactin assembly. AB - We have studied the effects of extrinsic environmental conditions on the conformation of surfactin, a heptapeptide biosurfactant from Bacillus subtilis, in aqueous solutions. It has been made clear that temperature, pH, Ca2+ ions and the synthetic nonionic surfactant hepta-ethylene glycol (C12E7) affect the conformation of surfactin in aqueous solutions. The beta-sheet formation reached a maximum at 40 degrees C both in presence and absence of (C12E7) and the nonionic surfactant enhances the beta-sheet formation even at 25 degrees C. Ca2 + induced the formation of alpha-helices and caused this transition at 0.3 mM with surfactin monomers or at 0.5 mM with surfactin micelles, but above these transition concentrations of Ca2+ beta-sheets were observed. In micellar solution the beta-sheet structure was stabilized at pH values below 7 or upon addition of Ca2+ in concentrations above 0.5 mM. Our results indicated that the bioactive conformation of surfactin is most likely the beta-sheets when the molecules are assembled in micelles. The beta-sheet structure in micelles could be retained by tuning the micelles. Surfactin micelles could be tuned in the bioactive conformation by manipulating pH, temperature, Ca2+ or (C12E7) concentrations in surfactin solutions. Our results strongly indicated that Ca2+ and other molecules (such as C12E7) may function as directing templates in the assembly and conformation of surfactin in micelles. Thus, we suggest environmental manipulation and template-aided micellation (TAM) as a new approach for preparing predesigned micelles, microemulsions or micro-spheres for specific application purposes. PMID- 9851373 TI - The hidden epidemic of cardiovascular disease. PMID- 9851374 TI - Evoked potentials not just to confirm hopelessness in anoxic brain injury. PMID- 9851375 TI - Appendicectomy and ulcerative colitis. PMID- 9851376 TI - Zero progress with hypochondriasis. PMID- 9851377 TI - Rubella gene sequencing as a clinician's tool. PMID- 9851378 TI - Learning a culture of respect for human rights. PMID- 9851379 TI - Blood pressure, cholesterol, and stroke in eastern Asia. Eastern Stroke and Coronary Heart Disease Collaborative Research Group. AB - BACKGROUND: Stroke is a major cause of death and disability in most populations of eastern Asia, and the incidence, particularly of haemorrhagic stroke, is generally higher than in western populations. This study aimed to assess the contributions of blood pressure and blood cholesterol concentrations to stroke risk in populations from eastern Asia. METHODS: The project included 13 cohorts from the People's Republic of China and five from Japan (124,774 participants, 837,214 person-years of observation). All 18 cohorts provided data on blood pressure and 12 (69,767 participants) provided data on cholesterol concentrations. Parametric and non-parametric analyses were done, with adjustments for several potential confounding factors. Analyses were based on estimated usual diastolic blood pressure and cholesterol concentration during follow-up, rather than baseline measurements, to avoid regression dilution bias. FINDINGS: Overall mean blood pressure was 124/78 mm Hg and mean cholesterol concentration was 4.5 mmol/L. 1798 strokes occurred; 751 (42%) were classified as haemorrhagic and 707 (39%) were confirmed by computed tomography or necropsy. Each 5 mm Hg lower usual diastolic blood pressure was associated with lower risk of non-haemorrhagic stroke (odds ratio 0.61 [95% CI 0.57-0.66]) and lower risk of haemorrhagic stroke (0.54 [0.50-0.58]). With decreasing cholesterol concentrations there were trends towards a decrease in risk of non-haemorrhagic stroke (odds ratio for 0.6 mmol/L decrease, 0.77 [0.57-1.06]) and an increase in risk of haemorrhagic stroke (1.27 [0.84-1.91]). Overall, there was no clear evidence of any interaction between cholesterol and diastolic blood pressure. INTERPRETATION: Blood pressure is an important determinant of stroke risk in eastern Asian populations, whereas cholesterol concentration is less important, affecting the proportions of stroke subtypes more than overall stroke numbers. The association between blood pressure and stroke seems stronger than in western populations; a population-wide reduction of 3 mm Hg in diastolic blood pressure should eventually decrease the number of strokes by about a third. PMID- 9851380 TI - Systematic review of early prediction of poor outcome in anoxic-ischaemic coma. AB - BACKGROUND: Studies to assess the prognostic value of early neurological and neurophysiological findings in patients with anoxic-ischaemic coma have not led to precise, generally accepted, prognostic rules. We did a systematic review of the relevant literature to assess whether such rules could be derived from the combined results of these studies. METHODS: From Medline and Embase databases we selected studies concerning patients older than 10 years with anoxic-ischaemic coma in which findings from early neurological examination, electroencephalogram (EEG), or somatosensory evoked potentials (SSEP) were related to poor outcome- defined as death or survival in a vegetative state. We selected variables with a specificity of 100% for poor outcome in all studies, and expressed the overall prognostic accuracy of these variables as pooled positive-likelihood ratios and as 95% CIs of the pooled false-positive test rates. FINDINGS: In 33 studies, 14 prognostic variables were studied, three of which had a specificity of 100%: absence of pupillary light reflexes on day 3 (pooled positive-likelihood ratio 10.5 [95% CI 2.1-52.4]; 95% CI pooled false-positive test rate 0-11.9%); absent motor response to pain on day 3 (16.8 [3.4-84.1]; 0-6.7%); and bilateral absence of early cortical SSEP within the first week (12.0 [5.3-27.6]; 0-2.0%). EEG recordings with an isoelectric or burst-suppression pattern had a specificity of 100% in five of six relevant studies (pooled positive-likelihood ratio 9.0 [2.5 33.1]; 95%CI pooled false-positive test rate 0.2-5.9%). These characteristics were present in 19%, 31%, 33%, and 33% of pooled patient populations, respectively. For the 11 SSEP studies, results did not significantly differ between studies in which the treating physicians were or were not masked from the test result, prospective and retrospective studies, studies with short and long follow-up periods, and studies with high or low overall poor outcome. INTERPRETATION: SSEP has the smallest CI of its pooled positive-likelihood ratio and its pooled false-positive test rate. Because evoked potentials are also the least susceptible to metabolic changes and drugs, recording of SSEP is the most useful method to predict poor outcome. PMID- 9851381 TI - Cohort study on circumcision of newborn boys and subsequent risk of urinary-tract infection. AB - BACKGROUND: A decrease in risk of urinary-tract infection is one of the most commonly given reasons for circumcision of newborn boys. Previous studies have reported rates of UTI to be 10-20 times higher in uncircumcised than in circumcised boys. This population-based cohort study followed neonates in Ontario, Canada, prospectively to study the relation between circumcision and subsequent UTI risk. METHODS: Eligible boys were born to residents of Ontario between April 1, 1993, and March 31, 1994. We used hospital discharge data to follow up boys until March 31, 1996. FINDINGS: Of 69,100 eligible boys, 30,105 (43.6%) were circumcised and 38,995 (56.4%) uncircumcised. 888 boys circumcised after the first month of life were excluded. 29,217 uncircumcised boys were matched to the remaining circumcised boys by date of birth. The 1-year probabilities of hospital admission for UTI were 1.88 per 1000 person-years of observation (83 cases up to end of follow-up) in the circumcised cohort and 7.02 per 1000 person-years (247 cases up to end of follow-up) in the uncircumcised cohort (p<0.0001). The estimated relative risk of admission for UTI by first-year follow-up indicated a significantly higher risk for uncircumcised boys than for circumcised boys (3.7 [2.8-4.9]). 195 circumcisions would be needed to prevent one hospital admission for UTI in the first year of life. INTERPRETATION: Although our findings support the notion that circumcision may protect boys from UTI, the magnitude of this effect may be less than previously estimated. PMID- 9851383 TI - Trial of oral miltefosine for visceral leishmaniasis. AB - BACKGROUND: There is no effective oral treatment for visceral leishmaniasis (kala azar), a disseminated intracellular protozoal infection that occurs worldwide. Miltefosine, an alkyl phospholipid developed as an oral antineoplastic agent, is active against visceral infection in animal models. We tested safety, tolerance, and efficacy of miltefosine in kala-azar. METHODS: Oral doses of miltefosine were given to six groups of five Indian men for 28 days: 50 mg every second day (group 1), 100 mg every second day (group 2), 100 mg/day (group 3), 150 mg/day (group 4), 200 mg/day (group 5), and 250 mg/day (group 6). Assessment for apparent cure- taken as an afebrile state with decreased spleen size and a splenic-aspirate parasite-density score of 0--was done on days 14 and 28. Definitive cure at 8 months required a parasite-free bone-marrow aspirate and no clinical evidence of relapse. FINDINGS: 21 of 30 patients were apparently cured on day 14. Transient episodes of vomiting and diarrhoea, were common during weeks 1-2 and were seen in 22 patients. Four other patients in groups 5 and 6 had miltefosine withdrawn after 7-10 days because of vomiting. One patient in group 6 developed renal insufficiency and severe diarrhoea and died on day 21. On day 28, all 29 remaining patients were apparently cured. By 8 months, seven of ten patients in groups 1 and 2 had relapsed; however, 18 of 19 patients treated daily (groups 3 6) appeared to be cured. Among the 21 definitive cures were the four patients treated for 10 days or less and 12 for whom previous therapy with pentavalent antimony had failed. INTERPRETATION: Treatment with miltefosine at 100-150 mg/day for 4 weeks has promise as an effective oral treatment of visceral leishmaniasis including antimony-resistant infection. PMID- 9851382 TI - Organochlorine exposure and risk of breast cancer. AB - BACKGROUND: Some organochlorine compounds may have weak oestrogenic effects and are, therefore, suspected of increasing the risk of breast cancer. We assessed prospectively the risk of breast cancer in relation to serum concentrations of several organochlorine compounds. METHODS: In 1976, serum samples from 7712 women were obtained from participants in the Copenhagen City Heart Study as part of physical examinations and interviews about lifestyle factors. During 17 years of follow-up, 268 women developed invasive breast cancer. Each woman with breast cancer was matched with two breast-cancer-free women from the remaining cohort. We analysed in 1996-97 the serum samples from 240 women with breast cancer and 477 controls. FINDINGS: Dieldrin was associated with a significantly increased dose-related risk of breast cancer (adjusted odds ratio 2.05 [95% CI 1.17-3.57], p for trend 0.01). Beta-hexachlorocyclohexane increased risk slightly but not significantly (p for trend 0.24). There was no overall association between risk of breast cancer and p,p'-dichlorodiphenyltrichloroethane or metabolites or for polychlorinated biphenyls. Exclusion of women with breast cancer diagnosed within 5 years of blood sampling strengthened the result for dieldrin, but did not affect the other results. INTERPRETATION: These findings support the hypothesis that exposure to xeno-oestrogens may increase the risk of breast cancer. PMID- 9851384 TI - Dipsticks and convulsions. PMID- 9851385 TI - Non-invasive imaging of arrhythmogenic left-ventricular myocardium after infarction. PMID- 9851386 TI - Subacute cutaneous lupus erythematosus associated with interferon beta-1a. PMID- 9851387 TI - Low birthweight and renal disease in Australian aborigines. PMID- 9851388 TI - Abnormal retinal vascularisation in preterm children as a general vascular phenomenon. PMID- 9851389 TI - Trimethoprim and fasting plasma homocysteine. PMID- 9851390 TI - Borna disease virus proteins in cerebrospinal fluid of patients with recurrent depression and multiple sclerosis. PMID- 9851391 TI - Homocysteine and folate in healthy east London Bangladeshis. PMID- 9851392 TI - Increased thrombotic vascular events after change of statin. PMID- 9851393 TI - Identification of intact peanut lectin in peripheral venous blood. PMID- 9851394 TI - Comparison of effectiveness of direct recording scale and dial scale. PMID- 9851395 TI - Myasthenic crisis after botulinum toxin. PMID- 9851396 TI - Gene therapy takes further steps into the clinic. PMID- 9851397 TI - Tune in early for best results with cochlear implants. PMID- 9851398 TI - One year on, Japan has yet to accept organ transplantation. PMID- 9851399 TI - Popular drama prompts interest in HIV in Japan. PMID- 9851400 TI - Vertigo. AB - Vertigo is a subtype of dizziness, which results from an imbalance within the vestibular system. This seminar focuses on three common presentations of vertigo: prolonged spontaneous vertigo, recurrent attacks of vertigo, and positional vertigo. The patient's history is usually the key to differentiation of peripheral and central causes of vertigo. The most common cause of vertigo, benign paroxysmal positional vertigo, can be cured with a simple positional manoeuvre. Other common causes of vertigo include vestibular neuritis, Meniere's syndrome, migraine, and vertebrobasilar insufficiency. Treatment should be directed at the underlying cause whenever possible, but antivertiginous and antiemetic drugs can suppress symptoms when a specific cause cannot be found. These drugs are generally not indicated for long-term daily use, however, since they may interfere with the normal compensation process. PMID- 9851401 TI - Bone-marrow donation by mentally incapable adults. PMID- 9851402 TI - Can we afford medical care for Alice C? PMID- 9851403 TI - Truth and reconciliation...and justice. PMID- 9851404 TI - Human rights in the USA: land of the free? PMID- 9851405 TI - Torture continues in Turkey: findings of new report. PMID- 9851406 TI - Epileptology of the first-seizure presentation. PMID- 9851407 TI - Epileptology of the first-seizure presentation. PMID- 9851408 TI - Epileptology of the first-seizure presentation. PMID- 9851409 TI - Epileptology of the first-seizure presentation. PMID- 9851410 TI - GMDP and psoriasis. PMID- 9851411 TI - Atrial fibrillation. PMID- 9851412 TI - Atrial fibrillation. PMID- 9851413 TI - Atrial fibrillation. PMID- 9851414 TI - Atrial fibrillation. PMID- 9851415 TI - Bone scintigraphy in screening of torture survivors. PMID- 9851416 TI - Measuring the burden of disease. PMID- 9851417 TI - Psittacosis in a rural Australian town. PMID- 9851418 TI - Magnesium sulphate in pre-eclampsia. PMID- 9851419 TI - Semen quality and fertility. PMID- 9851420 TI - Antenatal screening for Down's syndrome. PMID- 9851421 TI - Antenatal screening for Down's syndrome. PMID- 9851422 TI - Effect of witnessed resuscitation on bereaved relatives. PMID- 9851423 TI - Euthanasia legislation. PMID- 9851424 TI - 5300 years ago, the Ice Man used natural laxatives and antibiotics. PMID- 9851425 TI - Treating HIV/AIDS information overload. PMID- 9851426 TI - Saturdays are radio days for psychiatric patients in Argentina. PMID- 9851427 TI - The Nobel chronicles. 1929: Christiaan Eijkman (1858-1930) and Frederick Hopkins (1861-1947). PMID- 9851428 TI - Consequences of neonatal seizures in the rat: morphological and behavioral effects. AB - Whereas neonatal seizures are a predictor of adverse neurological outcome, there is controversy regarding whether seizures simply reflect an underlying brain injury or can cause damage. We subjected neonatal rats to a series of 25 brief flurothyl-induced seizures. Once mature the rats were compared with control littermates for spatial learning and activity level. Short-term effects of recurrent seizures on hippocampal excitation were assessed by using the intact hippocampus formation preparation and long-term effects by assessing seizure threshold. Brains were analyzed for neuronal loss, sprouting of granule cell axons (mossy fibers), and neurogenesis. Compared with controls, rats subjected to neonatal seizures had impaired learning and decreased activity levels. There were no differences in paired-pulse excitation or inhibition or duration of afterdischarges in the intact hippocampal preparation. However, when studied as adults, rats with recurrent flurothyl seizures had a significantly lower seizure threshold to pentylenetetrazol than controls. Rats with recurrent seizures had greater numbers of dentate granule cells and more newly formed granule cells than the controls. Rats with recurrent seizures also had sprouting of mossy fibers in CA3 and the supragranular region. Recurrent brief seizures during the neonatal period have long-term detrimental effects on behavior, seizure susceptibility, and brain development. PMID- 9851429 TI - Imaging epileptogenic tubers in children with tuberous sclerosis complex using alpha-[11C]methyl-L-tryptophan positron emission tomography. AB - Several reports have indicated that cortical resection is effective in alleviating intractable epilepsy in children with tuberous sclerosis complex (TSC). Because of the multitude of cortical lesions, however, identifying the epileptogenic tuber(s) is difficult and often requires invasive intracranial electroencephalographic (EEG) monitoring. As increased concentrations of serotonin and serotonin-immunoreactive processes have been reported in resected human epileptic cortex, we used alpha-[11C]methyl-L-tryptophan ([11C]AMT) positron emission tomography (PET) to test the hypothesis that serotonin synthesis is increased interictally in epileptogenic tubers in patients with TSC. Nine children with TSC and epilepsy, aged 1 to 9 years (mean, 4 years 1 month), were studied. All children underwent scalp video-EEG monitoring, PET scans of glucose metabolism and serotonin synthesis, and EEG monitoring during both PET studies. [11C]AMT scans were coregistered with magnetic resonance imaging and with glucose metabolism scans. Whereas glucose metabolism PET showed multifocal cortical hypometabolism corresponding to the locations of tubers in all 9 children, [11C]AMT uptake was increased in one tuber (n=3), two tubers (n=3), three tubers (n=1), and four tubers (n=1) in 8 of the 9 children. All other tubers showed decreased [11C]AMT uptake. Ictal EEG data available in 8 children showed seizure onset corresponding to foci of increased [11C]AMT uptake in 4 children (including 2 with intracranial EEG recordings). In 2 children, ictal EEG was nonlocalizing, and in 1 child there was discordance between the region of increased [11C]AMT uptake and the region of ictal onset on EEG. The only child whose [11C]AMT scan showed no regions of increased uptake had a left frontal seizure focus on EEG; however, at the time of his [11C]AMT PET scan, his seizures had come under control. [11C]AMT PET may be a powerful tool in differentiating between epileptogenic and nonepileptogenic tubers in patients with TSC. PMID- 9851430 TI - Adult polyglucosan body disease in Ashkenazi Jewish patients carrying the Tyr329Ser mutation in the glycogen-branching enzyme gene. AB - Adult polyglucosan body disease (APBD) is a late-onset, slowly progressive disorder of the nervous system caused by glycogen branching enzyme (GBE) deficiency in a subgroup of patients of Ashkenazi Jewish origin. Similar biochemical finding is shared by glycogen storage disease type IV (GSD IV) that, in contrast to APBD, is an early childhood disorder with primarily systemic manifestations. Recently, the GBE cDNA was cloned and several mutations were characterized in different clinical forms of GSD IV. To examine whether mutations in the GBE gene account for APBD, we studied 7 patients from five Jewish families of Ashkenazi ancestry. The diagnosis was based on the typical clinical and pathological findings, and supported by reduced GBE activity. We found that the clinical and biochemical APBD phenotype in all five families cosegregated with the Tyr329Ser mutation, not detected in 140 controls. As this mutation was previously identified in a nonprogressive form of GSD IV and was shown in expression studies to result in a significant residual GBE activity, present findings explain the late onset and slowly progressive course of APBD in our patients. We conclude that APBD represents an allelic variant of GSD IV, but the reason for the difference in primary tissue involvement must be established. PMID- 9851431 TI - Matrilineal inheritance of complex I dysfunction in a multigenerational Parkinson's disease family. AB - Recent data suggesting complex I dysfunction in Parkinson's disease (PD) arises from mitochondrial DNA (mtDNA) mutation does not conclusively answer whether the responsible genetic lesion is inherited (primary) or somatic (secondary). To address this question, we identified a family in which multiple members over three generations are affected with PD through exclusively maternal lines. Cytoplasmic hybrids (cybrids) were created for 15 family members over two generations by transferring each individual's mtDNA to mtDNA-depleted human neuroblastoma cells. Eight of the 15 cybrid lines contained mtDNA obtained from maternally descended family members and seven contained mtDNA from paternally descended family members. After 6 weeks of culture, cybrid cell lines were assayed for complex I activity and oxidative stress, and mitochondrial morphology was analyzed by electron microscopy. Compared with the cybrid lines containing mtDNA from paternal descendants, cybrid lines containing mtDNA from maternal descendants had lower complex I activity, increased reactive oxygen species production, increased radical scavenging enzyme activities, and more abnormal mitochondrial morphologic features. These findings were present in cybrid lines containing mtDNA from maternal descendants with PD as well as in currently asymptomatic young maternal descendants, and support a precedent for inherited mtDNA mutation in some persons with PD. PMID- 9851432 TI - Beta-trace protein in cerebrospinal fluid: a blood-CSF barrier-related evaluation in neurological diseases. AB - Beta-trace protein concentrations in cerebrospinal fluid (CSF) and serum from 113 patients with various neurological diseases and 65 controls were determined with a sensitive and specific immunonephelometric assay. In adult control patients, beta-trace concentrations were 14.6+/-4.6 mg/L in CSF and 0.46+/-0.13 mg/L in serum, that is, 32-fold higher in CSF. beta-trace levels in CSF correlated with age as well as with the albumin CSF/serum ratio (Q(Alb)), which is considered a measure for blood-CSF barrier function. The relationship between CSF beta-trace levels and elevated Q(Alb) values was studied in various neurological diseases with CSF protein increase. In spinal canal stenosis, CSF beta-trace (mean=29.5+/ 10.5 mg/L) correlated positively with increasing Q(Alb) values. In bacterial meningitis, CSF beta-trace (mean=8.7+/-3.9 mg/L) remained invariant to changes of Q(Alb) values. In Guillain-Barre syndrome, CSF beta-trace (mean=14.4+/-6.8 mg/L) was below the Q(Alb)-dependent reference range. In multiple sclerosis and viral meningoencephalitis, beta-trace levels were within the reference range. Beta trace concentration in CSF can be used in conjunction with Q(AlB) to distinguish between different neurological pathologies associated with CSF protein increase. PMID- 9851433 TI - Familial partial epilepsy with variable foci: a new partial epilepsy syndrome with suggestion of linkage to chromosome 2. AB - Familial partial epilepsy with variable foci (FPEVF) joins the recently recognized group of inherited partial epilepsies. We describe an Australian family with 10 individuals with partial seizures over four generations. Detailed electroclinical studies were performed on all affected and 17 clinically unaffected family members. The striking finding was that the clinical features of the seizures and interictal electroencephalographic foci differed among family members and included frontal, temporal, occipital, and centroparietal seizures. Mean age of seizure onset was 13 years (range, 0.75-43 years). Two individuals without seizures had epileptiform abnormalities on electroencephalographic studies. Penetrance of seizures was 62%. A genome-wide search failed to demonstrate definitive linkage, but a suggestion of linkage was found on chromosome 2q with a LOD score of 2.74 at recombination fraction of zero with the marker D2S133. FPEVF differs from the other inherited partial epilepsies where partial seizures in different family members are clinically similar. The inherited nature of this new syndrome may be overlooked because of relatively low penetrance and because of the variability in age at onset and electroclinical features between affected family members. PMID- 9851434 TI - Aicardi-Goutieres syndrome: an update and results of interferon-alpha studies. AB - Twenty-seven patients with familial encephalopathy with calcification of the basal ganglia and chronic cerebrospinal fluid (CSF) lymphocytosis (Aicardi Goutieres syndrome) are reviewed. In 19 children, the onset was within the first 4 months of life. Most patients had normal head circumference at birth, but 21 developed microcephaly between 3 and 12 months. Neuroimaging showed severe and progressive brain atrophy in all patients. The extent and intensity of the calcification was variable even in the same sibship. CSF lymphocytosis persisted beyond 12 months of age in 7 children. High levels of interferon-alpha were found in serum and CSF in 14 patients. The higher CSF levels suggest intrathecal synthesis. Tubuloreticular inclusions related to the presence of interferon were found in 4 additional children. The 19 patients still alive (6 older than 10 years) are profoundly disabled. However, the syndrome may present with individual variations in severity, rapidity of evolution, and imaging features. Neuropathological examination in 2 patients failed to detect significant inflammatory lesions and showed only foci of necrosis and wide-spread demyelination. This study supports an autosomal recessive inheritance for this syndrome. The high level of interferon-alpha is not explained but may play a role in the pathogenesis of the disorder. PMID- 9851435 TI - Risk of unprovoked seizure after acute symptomatic seizure: effect of status epilepticus. AB - We asked whether acute symptomatic status epilepticus (SE) increases the risk for subsequent unprovoked seizure compared with less prolonged acute symptomatic seizure. We also explored whether the risk of unprovoked seizure differs by cause. We ascertained all first episodes of acute symptomatic seizure among residents of Rochester, Minnesota, through the Rochester Project's records linkage system. Information was collected on seizure duration, age, sex, cause, and subsequent unprovoked seizure. At 10 years of follow-up, the risk of unprovoked seizure was 41% for those with acute symptomatic seizure with SE and 13% for those without SE. Controlling for age, sex, and cause, SE increased the risk for subsequent unprovoked seizure 3.3-fold (95% confidence interval, 1.8 6.1) compared with brief acute symptomatic seizures. Among patients with SE, the risk of unprovoked seizure was increased 18.8-fold for patients with anoxic encephalopathy, 7.1-fold for patients with a structural cause, 3.6-fold for patients with a metabolic cause. The increased risk for unprovoked seizure after SE compared with shorter seizures may be due to SE being a marker for severity of injury, damage caused by SE, or a biological substrate associated with the tendency to experience SE. PMID- 9851436 TI - Neuromuscular blockade by IgG antibodies from patients with Guillain-Barre syndrome: a macro-patch-clamp study. AB - Guillain-Barre syndrome (GBS) is often associated with serum antibodies to glycoconjugates such as GM1 and GQ1b. The pathogenic role of these antibodies and other serum factors has not yet been clarified. We have investigated the effect of serum, plasma filtrate, and highly purified IgG and IgM from 10 patients with typical GBS on motor nerve terminals in the mouse hemidiaphragm. Quantal endplate currents were recorded by means of a perfused macro-patch-clamp electrode. The plasma filtrate of all GBS patients led to a 5- to 20-fold reduction of evoked quantal release within 7 to 15 minutes of continuous superfusion. In 4 patients, the amplitudes of single quanta were clearly reduced (by 10-66% of control values), indicating an additional postsynaptic action. Blocking effects could be reversed to a variable degree within 15 to 18 minutes after washout. Purified IgG was as effective as native serum, whereas a purified GBS IgM fraction did not block transmission. Sera from convalescent patients and IgG from healthy subjects were without blocking effect. The effects were complement independent and there was no link to the presence (in 6 patients) or absence (in 4 patients) of detectable antibodies to GM1 or GQ1b. In GBS, antibodies to an undetermined antigen depress the presynaptic transmitter release and, in some cases, the activation of postsynaptic channels. We suggest that weakness in the acute stage of GBS may be caused in part by circulating antibodies. PMID- 9851437 TI - Remodeling of neuronal circuitries in human temporal lobe epilepsy: increased expression of highly polysialylated neural cell adhesion molecule in the hippocampus and the entorhinal cortex. AB - Neuronal loss and axonal sprouting are the most typical histopathological findings in the hippocampus of patients with drug-refractory temporal lobe epilepsy (TLE). It is under dispute, however, whether remodeling of neuronal circuits is a continuous process or whether it occurs only during epileptogenesis. Also, little is known about the plasticity outside of the hippocampus. We investigated the immunoreactivity of the highly polysialylated neural cell adhesion molecule (PSA-NCAM) in the surgically removed hippocampus and the entorhinal cortex of patients with drug-refractory TLE (n=25) and autopsy controls (n=7). Previous studies have shown that the expression of PSA-NCAM is associated with the induction of synaptic plasticity, neurite outgrowth, neuronal migration, and events requiring remodeling or repair of tissue. In patients with TLE, the optical density (OD) of punctate PSA-NCAM immunoreactivity was increased both in the inner and outer molecular layers of the dentate gyrus, compared with controls. The intensity of PSA-NCAM immunoreactivity in the inner molecular layer correlated with the duration of epilepsy, severity of hippocampal neuronal loss, density of mossy fiber sprouting, and astrogliosis. In TLE patients with only mild neuronal loss in the hippocampus, the density of infragranular immunopositive neurons was increased twofold compared with controls, whereas in TLE patients with severe neuronal loss, the infragranular PSA-NCAM-positive cells were not present. In the hilus, the somata and tortuous dendrites of some surviving neurons were intensely stained in TLE. PSA-NCAM immunoreactivity was also increased in CA1 and in layer II of the rostral entorhinal cortex, where immunopositive neurons were surrounded by PSA-NCAM-positive fibers and puncta. Our data provide evidence that synaptic reorganization is an active process in human drug-refractory TLE. Moreover, remodeling is not limited to the dentate gyrus, but also occurs in the CA1 subfield and the entorhinal cortex. PMID- 9851438 TI - Molecular genetic analysis of a novel Parkin gene in Japanese families with autosomal recessive juvenile parkinsonism: evidence for variable homozygous deletions in the Parkin gene in affected individuals. AB - Autosomal recessive juvenile parkinsonism (AR-JP) is a distinct clinical and genetic entity characterized by selective degeneration of nigral dopaminergic neurons and young-onset parkinsonism with remarkable response to levodopa. Recently, we mapped the gene locus for AR-JP to chromosome 6q25.2-q27 by linkage analysis and we identified a novel large gene, Parkin, consisting of 12 exons from this region; mutations of this gene were found to be the cause of AR-JP in two families. Now we report results of extensive molecular analysis on 34 affected individuals from 18 unrelated families with AR-JP. We found four different homozygous intragenic deletional mutations, involving exons 3 to 4, exon 3, exon 4, and exon 5 in 10 families (17 affected individuals). In addition to the exonic deletions, we identified a novel one-base deletion involving exon 5 in two families (2 affected individuals). All mutations so far found were deletional types in which large exonic deletion accounted for 50% (17 of 34) and the one-base deletion accounted for 6% (2/34); in the remaining, no homozygous mutations were found in the coding regions. Our findings indicate that loss of function of the Parkin protein results in the clinical phenotype of AR-JP and that subregions between introns 2 and 5 of the Parkin gene are mutational hot spots. PMID- 9851439 TI - Reduction in HTLV-I proviral load and spontaneous lymphoproliferation in HTLV-I associated myelopathy/tropical spastic paraparesis patients treated with humanized anti-Tac. AB - Human T-lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a neurological disease that results from an interaction of retroviral infection and immune activation. In this study, five doses (1 mg/kg) of humanized anti-Tac antibody were administered to 9 HAM/TSP patients at weeks 0, 2, 6, 10, and 14. Preliminary immunological studies on HAM/TSP patients treated with humanized anti-Tac indicate that there is a selective down regulation of activated T cells and a decrease in the HTLV-I viral load in peripheral blood lymphocytes, most likely through the selective removal of HTLV-I infected, activated CD4+ lymphocytes. PMID- 9851440 TI - Vigabatrin increases human brain homocarnosine and improves seizure control. AB - Homocarnosine, a dipeptide of gamma-aminobutyric acid (GABA) and histidine, is thought to be an inhibitory neuromodulator synthesized in subclasses of GABAergic neurons. Homocarnosine is present in human brain in greater amounts (0.4-1.0 micromol/g) than in other animals. The antiepileptic drug vigabatrin increases human cerebrospinal fluid homocarnosine linearly with daily dose. By using 1H nuclear magnetic resonance spectroscopy, serial occipital lobe GABA and homocarnosine concentrations were measured in 11 patients started on vigabatrin. Daily low-dose (2 g) vigabatrin increased both homocarnosine and GABA. Larger doses of vigabatrin (4 g) further increased homocarnosine but changed GABA levels minimally. Seizure control improved with increasing homocarnosine and GABA concentrations. Patients whose seizure control improved with the addition of vigabatrin had higher mean homocarnosine, but the same mean GABA concentrations, than those whose seizure control did not improve. Increased homocarnosine may contribute to improved seizure control. PMID- 9851441 TI - Bilateral high-frequency stimulation of the internal globus pallidus in advanced Parkinson's disease. AB - We report here the results of an open prospective study in 9 patients suffering from severe Parkinson's disease with on/off fluctuations and restricted off period mobility, who underwent bilateral implantation of stimulating electrodes in the internal pallidum. At 3-month follow-up, the total Unified Parkinson's Disease Rating Scale (UPDRS) motor score in the medication-off state was reduced from 54.1+/-14.8 to 23.9+/-11.7 (44.2%) when stimulation was turned on. Comparison of UPDRS subscores revealed significant improvements for tremor, rigidity, bradykinesia, gait and posture, and dyskinesias. The results of the clinical scoring could be confirmed by significant changes in the quantitative assessment of hand function and walking. Bilateral pallidal stimulation reduced the amount and severity of on/off fluctuations. Additional follow-up at 6 months (n=6), 9 months (n=6), and 12 months (n=4) did not show a decline in effectiveness of stimulation. There was no permanent morbidity associated with the procedure. A subtle reduction of verbal fluency, which was not evident to the patients, was the only cognitive side effect of the procedure in neuropsychological testing. Chronic bilateral high-frequency stimulation of the internal pallidum seems to be a neurologically safe and highly effective treatment for "off" symptoms, dyskinesias, and motor fluctuations in advanced stages of Parkinson's disease. PMID- 9851442 TI - The mitochondrial DNA A8344G mutation in Leigh syndrome revealed by analysis in paraffin-embedded sections: revisiting the past. AB - In 1975, we presented the results of a study on a family with a constellation of features that included a chronic spinocerebellar syndrome, neuropathologically proven Leigh syndrome, and sudden death in infancy or childhood affecting several members over three generations. Inheritance was thought to be autosomal dominant. Twenty years later, we reinterpreted the inheritance pattern as maternal. Mitochondrial DNA (mtDNA) extracted from paraffin-embedded brain samples from the proband revealed the A8344G myoclonic epilepsy and ragged-red fiber (MERRF) mutation as the molecular basis for this multifaceted neurological syndrome. This re-evaluation of archival material is an instructive example of "medical archeopathology." PMID- 9851443 TI - The Glu318Gly mutation of the presenilin-1 gene does not necessarily cause Alzheimer's disease. AB - In early-onset familial Alzheimer's disease (AD) pathogenic mutations have been found in the amyloid precursor protein (APP) gene and in the presenilin (PS)-1 and PS-2 genes. We screened for mutations in these genes in 20 patients with familial AD from the Finnish population. In addition, we sampled 41 sporadic AD patients and 59 controls to test for mutations identified in our familial AD cases. We detected an A-to-G transition in the PS-1 gene, resulting in a glutamic acid (Glu)-to-glycine (Gly) substitution at codon 318 in 2 familial and 2 sporadic AD patients. The Glu318Gly mutation has previously been reported to cause AD. We also found the Glu318Gly mutation in 4 healthy aged controls (range, 74-87 years). We thus conclude that the mutation is most likely a rare polymorphism not related to AD. PMID- 9851444 TI - Muscle fiber immaturity and inactivity reduce myonecrosis in Duchenne muscular dystrophy. AB - We report on the first case of X-linked recessive myotubular myopathy (MTM1) coinciding with Duchenne muscular dystrophy (DMD). The muscle biopsy specimens of the patient show only the characteristic findings of MTM1, without the findings of DMD. We theorize that this was caused by the muscle fiber immaturity and inactivity. PMID- 9851445 TI - A dystrophin missense mutation showing persistence of dystrophin and dystrophin associated proteins yet a severe phenotype. AB - A muscle biopsy from an X-linked muscular dystrophy pedigree showed normal dystrophin and dystrophin-associated proteins. Linkage to multiple markers within the dystrophin gene (LOD=2.7, theta=0) indicated a primary dystrophinopathy. Sequencing of the entire dystrophin RNA revealed a single missense mutation (D3335H) in the unique carboxyl-terminal domain. This is the first report showing that a relatively severe dystrophinopathy can occur despite the correct localization of dystrophin and dystrophin-associated proteins. PMID- 9851446 TI - Utility of anti-Hu antibodies in the diagnosis of paraneoplastic sensory neuropathy. AB - Anti-Hu antibodies (Hu-Abs) were positive in 40 patients with paraneoplastic sensory neuropathy (PSN) and in 1 patient with idiopathic sensory neuropathy in a series of 126 patients who presented with clinical features suggestive of PSN. The specificity of Hu-Abs was 99% and the sensitivity was 82%. Nine (18%) PSN patients were Hu-Ab-negative, and their sera did not harbor other specific anti neuronal or anti-ganglioside antibodies. Small cell lung carcinoma (SCLC) was the leading neoplasm in the Hu-Ab-positive (79%) and Hu-Ab-negative (44%) groups. This study confirms the value of Hu-Abs for the diagnosis of PSN and SCLC and also emphasizes that in patients with possible PSN, the absence of Hu-Abs does not exclude cancer, particularly in those patients with risk factors for SCLC. PMID- 9851448 TI - Susceptibility to multiple sclerosis and the immunoglobulin heavy chain gene cluster. PMID- 9851447 TI - Increased circulating antiganglioside antibodies in primary and secondary progressive multiple sclerosis. AB - Plasma samples from 70 patients with multiple sclerosis (MS), 41 patients with other neurological diseases (OND), and 38 healthy subjects were examined for antibodies against gangliosides GM1, GM3, GD1a, GD1b, and GD3 using enzyme-linked immunosorbent assays. The percentages of subjects with increased anti-GM3 responses were significantly higher in the primary progressive MS (56.3%) and secondary progressive MS (42.9%) groups than in the relapsing-remitting MS (2.9%), healthy subject (2.6%), and OND (14.6%) groups. Elevated antiganglioside antibodies may be secondary to axonal damage or may be a cause of axonal damage and accumulating disability in progressive MS. In either case, they may serve as a marker of axonal damage in MS. PMID- 9851449 TI - Questionable role of adult-onset focal dystonia among sporadic dystonia patients. PMID- 9851450 TI - Missense mutation E318G of the presenilin-1 gene appears to be a nonpathogenic polymorphism. PMID- 9851451 TI - Prenatal white matter injury. PMID- 9851452 TI - Aging glia may not protect neurons. PMID- 9851453 TI - Reproductive medicine experts till an increasingly fertile field. PMID- 9851454 TI - Getting religion seen as help in being well. PMID- 9851455 TI - Do-it-yourself report on patient privacy. PMID- 9851456 TI - From the Centers for Disease Control and Prevention. Assessment of infant sleeping position--selected states, 1996. PMID- 9851457 TI - From the Centers for Disease Control and Prevention. Self-reported use of mammography and insurance status among women aged > or =40 years--United States, 1991-1992 and 1996-1997. PMID- 9851458 TI - A piece of my mind. Alton Small. PMID- 9851459 TI - The changing treatment paradigm for acute otitis media. PMID- 9851460 TI - The changing treatment paradigm for acute otitis media. PMID- 9851461 TI - Screening for alcoholism using CAGE. Cut down, Annoyed, Guilty, Eye opener. PMID- 9851462 TI - An even closer look at therapeutic touch. PMID- 9851463 TI - An even closer look at therapeutic touch. PMID- 9851464 TI - An even closer look at therapeutic touch. PMID- 9851465 TI - An even closer look at therapeutic touch. PMID- 9851466 TI - An even closer look at therapeutic touch. PMID- 9851467 TI - An even closer look at therapeutic touch. PMID- 9851468 TI - An even closer look at therapeutic touch. PMID- 9851469 TI - An even closer look at therapeutic touch. PMID- 9851470 TI - An even closer look at therapeutic touch. PMID- 9851471 TI - An even closer look at therapeutic touch. PMID- 9851472 TI - An even closer look at therapeutic touch. PMID- 9851473 TI - An even closer look at therapeutic touch. PMID- 9851474 TI - Endocarditis prophylaxis for transrectal prostatic biopsy. PMID- 9851475 TI - Bartenders' respiratory health after establishment of smoke-free bars and taverns. AB - CONTEXT: The association between environmental tobacco smoke (ETS) exposure and respiratory symptoms has not been well established in adults. OBJECTIVE: To study the respiratory health of bartenders before and after legislative prohibition of smoking in all bars and taverns by the state of California. DESIGN: Cohort of bartenders interviewed before and after smoking prohibition. SETTING AND PARTICIPANTS: Bartenders at a random sample of bars and taverns in San Francisco. MAIN OUTCOME MEASURES: Interviews assessed respiratory symptoms, sensory irritation symptoms, ETS exposure, personal smoking, and recent upper respiratory tract infections. Spirometric assessment included forced expiratory volume in 1 second (FEV1) and forced vital capacity (FVC) measurements. RESULTS: Fifty-three of 67 eligible bartenders were interviewed. At baseline, all 53 bartenders reported workplace ETS exposure. After the smoking ban, self-reported ETS exposure at work declined from a median of 28 to 2 hours per week (P<.001). Thirty-nine bartenders (74%) initially reported respiratory symptoms. Of those symptomatic at baseline, 23 (59%) no longer had symptoms at follow-up (P<.001). Forty-one bartenders (77%) initially reported sensory irritation symptoms. At follow-up, 32 (78%) of these subjects had resolution of symptoms (P<.001). After prohibition of workplace smoking, we observed improvement in mean FVC (0.189 L; 95% confidence interval [CI], 0.082-0.296 L; 4.2% change) and, to a lesser extent, mean FEV1 (0.039 L; 95% CI, -0.030 to 0.107 L; 1.2% change). Complete cessation of workplace ETS exposure (compared with continued exposure) was associated with improved mean FVC (0.287 L; 95% CI, 0.088-0.486; 6.8% change) and mean FEV1 (0.142 L; 95% CI, 0.020-0.264 L; 4.5% change), after controlling for personal smoking and recent upper respiratory tract infections. CONCLUSION: Establishment of smoke-free bars and taverns was associated with a rapid improvement of respiratory health. PMID- 9851476 TI - Declining blood lead levels and changes in cognitive function during childhood: the Port Pirie Cohort Study. AB - CONTEXT: Many studies have found a significant inverse association between early exposure to environmental lead and cognitive function in childhood. Whether these effects are reversible when exposure is reduced is not clear. OBJECTIVE: To assess the reversibility of the apparent effects of lead on cognitive abilities in early childhood by testing whether declines in blood lead concentrations beyond the age of 2 years are associated with improvements in cognition. SETTING: Urban and rural communities surrounding a large lead smelter in Port Pirie, South Australia. PARTICIPANTS: A total of 375 children followed up from birth to the age of 11 to 13 years. DESIGN: Long-term prospective cohort study. MAIN OUTCOME MEASURES: The Bayley Mental Development Index at age 2 years, the McCarthy General Cognitive Index at age 4 years, and IQs from the Wechsler Intelligence Scale (revised version) at ages 7 and 11 to 13 years. RESULTS: Mean blood lead concentrations in the children decreased from 1.02 pmol/L (21 .2 microg/dL) at age 2 years to 0.38 micromol/L (7.9 microg/dL) at age 1 1 to 13 years, but cognitive scores in children whose blood lead concentration declined most were generally not improved relative to the scores of children whose blood lead levels declined least. Changes in IQ and declines in blood lead levels that occurred between the ages of 7 and 11 to 13 years (r= 0.12, P= .09) suggested slightly better cognition among children whose blood lead levels declined most. CONCLUSION: The cognitive deficits associated with exposure to environmental lead in early childhood appear to be only partially reversed by a subsequent decline in blood lead level. PMID- 9851477 TI - High-altitude cerebral edema evaluated with magnetic resonance imaging: clinical correlation and pathophysiology. AB - CONTEXT: Because of its onset in generally remote environments, high-altitude cerebral edema (HACE) has received little scientific attention. Understanding the pathophysiology might have implications for prevention and treatment of both this disorder and the much more common acute mountain sickness. OBJECTIVES: To identify a clinical imaging correlate for HACE and determine whether the edema is primarily vasogenic or cytotoxic. DESIGN: Case-comparison study. SETTING: Community hospitals accessed by helicopter from mountains in Colorado and Alaska. PATIENTS: A consecutive sample of 9 men with HACE, between 18 and 35 years old, 8 of whom also had pulmonary edema, were studied after evacuation from high altitude locations; 5 were mountain climbers and 4 were skiers. The control group, matched for age, sex, and altitude exposure, consisted of 3 subjects with high-altitude pulmonary edema only and 3 who had been entirely well at altitude. Four patients with HACE were available for follow-up imaging after complete recovery. MAIN OUTCOME MEASURES: Magnetic resonance imaging (MRI) of the brain during acute, convalescent, and recovered phases of HACE, and once in controls, immediately after altitude exposure. RESULTS: Seven of the 9 patients with HACE showed intense T2 signal in white matter areas, especially the splenium of the corpus callosum, and no gray matter abnormalities. Control subjects demonstrated no such abnormalities. All patients completely recovered; in the 4 available for follow-up MRI, the changes had resolved entirely. CONCLUSIONS: We conclude that HACE is characterized on MRI by reversible white matter edema, with a predilection for the splenium of the corpus callosum. This finding provides a clinical imaging correlate useful for diagnosis. It also suggests that the predominant mechanism is vasogenic (movement of fluid and protein out of the vascular compartment) and, thus, that the blood-brain barrier may be important in HACE. PMID- 9851478 TI - Improved prognosis of thoracic aortic aneurysms: a population-based study. AB - CONTEXT: Managing thoracic aortic aneurysms identified incidentally by increased use of computed tomography, echocardiography, and magnetic resonance imaging is problematic, especially in the elderly. OBJECTIVE: To ascertain whether the previously reported poor prognosis for individuals with thoracic aortic aneurysms has changed with better medical therapies and improved surgical techniques that can now be applied to aneurysm management. DESIGN: Population-based cohort study. SETTING AND PATIENTS: All 133 patients with the diagnosis of degenerative thoracic aortic aneurysms among Olmsted County, Minnesota, residents between 1980 and 1994 compared with a previously reported cohort of similar patients between 1951 and 1980. MAIN OUTCOME MEASURES: The primary clinical end points were incidence, cumulative rupture risk, rupture risk as a function of aneurysm size, and survival. RESULTS: In contrast to abdominal aortic aneurysms, for which men are affected predominately, 51% of thoracic aortic aneurysms were identified in women who were considerably older at recognition than men (mean age, 75.9 vs 62.8 years, respectively; P= .01). The overall incidence rate of 10.4 per 100000 person-years (95% confidence interval [CI], 8.6-12.2) between 1980 and 1994 was more than 3-fold higher than the rate from 1951 to 1980. The cumulative risk of rupture was 20% after 5 years. Seventy-nine percent of ruptures occurred in women (P= .01). The 5-year risk of rupture as a function of aneurysm size at recognition was 0% for aneurysms less than 4 cm in diameter, 16% (95% CI, 4%-28%) for those 4 to 5.9 cm, and 31% (95% CI, 5%-56%) for aneurysms 6 cm or more. Overall 5-year survival improved to 56% (95% CI, 48%-66%) between 1980 and 1994 compared with only 19% between 1951 and 1980 (P<.01). CONCLUSIONS: In this population, elderly women represent an increasing portion of all patients with clinically recognized thoracic aortic aneurysms and constitute the majority of patients whose aneurysm eventually ruptures. Overall survival for thoracic aortic aneurysms has improved significantly in the past 15 years. PMID- 9851479 TI - Aspirin and risk of hemorrhagic stroke: a meta-analysis of randomized controlled trials. AB - CONTEXT: Aspirin has been widely used to prevent myocardial infarction and ischemic stroke but some studies have suggested it increases risk of hemorrhagic stroke. OBJECTIVE: To estimate the risk of hemorrhagic stroke associated with aspirin treatment. DATA SOURCES: Studies were retrieved using MEDLINE (search terms, aspirin, cerebrovascular disorders, and stroke), bibliographies of the articles retrieved, and the authors' reference files. STUDY SELECTION: All trials published in English-language journals before July 1997 in which participants were randomized to aspirin or a control treatment for at least 1 month and in which the incidence of stroke subtype was reported. DATA EXTRACTION: Information on country of origin, sample size, duration, study design, aspirin dosage, participant characteristics, and outcomes was abstracted independently by 2 authors who used a standardized protocol. DATA SYNTHESIS: Data from 16 trials with 55462 participants and 108 hemorrhagic stroke cases were analyzed. The mean dosage of aspirin was 273 mg/d and mean duration of treatment was 37 months. Aspirin use was associated with an absolute risk reduction in myocardial infarction of 137 events per 10000 persons (95% confidence interval [CI], 107 167; P<.001) and in ischemic stroke, a reduction of 39 events per 10000 persons (95% CI, 17-61; P<.001). However, aspirin treatment was also associated with an absolute risk increase in hemorrhagic stroke of 12 events per 10000 persons (95% CI, 5-20; P<.001). This risk did not differ by participant or study design characteristics. CONCLUSIONS: These results indicate that aspirin therapy increases the risk of hemorrhagic stroke. However, the overall benefit of aspirin use on myocardial infarction and ischemic stroke may outweigh its adverse effects on risk of hemorrhagic stroke in most populations. PMID- 9851480 TI - Effective medical treatment of opiate addiction. National Consensus Development Panel on Effective Medical Treatment of Opiate Addiction. AB - OBJECTIVE: To provide clinicians, patients, and the general public with a responsible assessment of the effective approaches to treat opiate dependence. PARTICIPANTS: A nonfederal, nonadvocate, 12-member panel representing the fields of psychology, psychiatry, behavioral medicine, family medicine, drug abuse, epidemiology, and the public. In addition, 25 experts from these same fields presented data to the panel and a conference audience of 600. Presentations and discussions were divided into 3 phases over 2 1/2 days: (1) presentations by investigators working in the areas relevant to the consensus questions during a 2 day public session; (2) questions and statements from conference attendees during open discussion periods that are part of the public session; and (3) closed deliberations by the panel during the remainder of the second day and morning of a third day. The conference was organized and supported by the Office of Medical Applications of Research, National Institutes of Health. EVIDENCE: The literature was searched through MEDLINE and other National Library of Medicine and online databases from January 1994 through September 1997 and an extensive bibliography of 941 references was provided to the panel and the conference audience. Experts prepared abstracts for their presentations as speakers at the conference with relevant citations from the literature. Scientific evidence was given precedence over clinical anecdotal experience. CONSENSUS PROCESS: The panel, answering predefined questions, developed its conclusions based on the scientific evidence presented in open forum and the scientific literature. The panel composed a draft statement that was read in its entirety and circulated to the experts and the audience for comment. Thereafter, the panel resolved conflicting recommendations and released a revised statement at the end of the conference. The panel finalized the revisions within a few weeks after the conference. The draft statement was made available on the World Wide Web immediately following its release at the conference and was updated with the panel's final revisions. CONCLUSIONS: Opiate dependence is a brain-related medical disorder that can be effectively treated with significant benefits for the patient and society, and society must make a commitment to offer effective treatment for opiate dependence to all who need it. All persons dependent on opiates should have access to methadone hydrochloride maintenance therapy under legal supervision, and the US Office of National Drug Control Policy and the US Department of Justice should take the necessary steps to implement this recommendation. There is a need for improved training for physicians and other health care professionals. Training to determine diagnosis and treatment of opiate dependence should also be improved in medical schools. The unnecessary regulations of methadone maintenance therapy and other long-acting opiate agonist treatment programs should be reduced, and coverage for these programs should be a required benefit in public and private insurance programs. PMID- 9851481 TI - Should physicians accept gifts from patients? PMID- 9851482 TI - Exposure to environmental tobacco smoke: identifying and protecting those at risk. PMID- 9851483 TI - Individualizing aspirin therapy for prevention of cardiovascular events. PMID- 9851484 TI - Updating protections for human subjects involved in research. Project on Informed Consent, Human Research Ethics Group. AB - For decades, all federally funded research involving human subjects has been subject to regulations that require the informed consent of the subject and oversight by the local institution. These regulations last underwent major revision in 1981 and have remained unchanged despite significant changes in the nature of clinical science, the financial sources of research support, and the institutional environment in which clinical research is conducted. In the intervening years, doubt has evolved as to whether the regulations currently in place adequately protect the welfare and rights of research subjects in today's clinical research environment and whether the costs, in terms of time, bureaucracy, and delay, are justified by the level of protection afforded. The Human Research Ethics Group, administered by the Center for Bioethics at the University of Pennsylvania Health System, extensively reviewed the status of existing human subjects protections with the aim of making recommendations to improve and reform the regulations. Here, we present recommendations constituting a consensus of the group members for reform in 3 key areas: protecting subject populations with special needs and vulnerabilities, oversight by institutional review boards, and regulatory policy. PMID- 9851485 TI - Testosterone replacement therapy for HIV-related wasting: abstract and commentary. PMID- 9851486 TI - A new HIV-1 group: abstract and commentary. PMID- 9851487 TI - JAMA patient page: secondhand smoke. PMID- 9851488 TI - Vocal mechanics as the basis for phonosurgery. AB - OBJECTIVE: A number of modifications in laryngoplastic phonosurgery have recently been proposed. This report is intended to clarify the concept on which the surgery should be based, vocal mechanics, for further rational development of the surgery. STUDY DESIGN: The results of various previous surgeries were compared. In an attempt to elucidate what makes the difference in the results, simulation of voice production was conducted with the use of excised larynges. METHODS: Excised larynges were mounted on a tube so as to be blown from below. Experimental variables in the model included the glottal area initially set, stiffness of the vocal folds, and subglottal pressure. The conditions under which the voice became hoarse were examined. RESULTS: The voice became hoarse under the following major conditions: 1) the initial glottal area exceeded a certain value, 2) stiffness was too high, and 3) the glottis was too tightly closed. Clinical representation for each condition was made, particularly for excessively tight glottal closure such as in spasmodic dysphonia. A new type of surgical treatment for spasmodic dysphonia, lateralization thyroplasty, was briefly reported, which restored the voice to normal without recurrence for 1 year at the time of this writing. CONCLUSIONS: A potential new type of laryngoplastic phonosurgery should be conformed to the mechanics of voice production. In treating dysphonia, it is often necessary to switch from etiologic or radical treatment, if infertile, to symptomatic treatment instead, at the level of mechanics. PMID- 9851489 TI - Acoustic and perceptual evaluation of laryngeal reinnervation by ansa cervicalis transfer. AB - OBJECTIVES/HYPOTHESIS: To evaluate the acoustic and perceptual results of laryngeal reinnervation with ansa cervicalis to recurrent laryngeal nerve anastomosis. STUDY DESIGN: Retrospective study of voice samples from 12 patients with unilateral recurrent laryngeal nerve paralysis, treated with ansa cervicalis to recurrent laryngeal nerve anastomosis. Samples were recorded before surgery and at least 8 months after surgery. METHODS: The samples were subjected to several acoustic analyses sensitive to paralytic dysphonia, including cepstral peak prominence, noise-to-harmonics ratio, and measures of frequency and amplitude perturbation. The voice samples from the patients were randomized with age- and sex-matched samples from normal subjects and judged by trained listeners for overall dysphonia, roughness, breathiness, asthenia, and strain. The preoperative and postoperative results were compared statistically, and the postoperative results were compared with the matched normal subjects. RESULTS: As a group, the patients showed improvement (P < .05) in cepstral peak prominence, frequency perturbation, and perceptual judgments of overall dysphonia, breathiness, and asthenia. The best results occurred in patients with isolated vocal fold paralysis. The postoperative group as a whole did not improve to the level seen in matched normals. Suboptimal results were seen primarily in patients with untreated laryngeal or extralaryngeal pathology beyond the laryngeal paralysis. CONCLUSIONS: These data suggest that laryngeal reinnervation has the potential to bring about a return to normal or near-normal voice in patients with isolated unilateral vocal fold paralysis. PMID- 9851490 TI - Videostroboscopy of the pharyngoesophageal segment in total laryngectomees. AB - OBJECTIVES: The reconstructed pharyngoesophageal segment (PES) serves as the neoglottis following total laryngectomy, as it provides the source of vibration for production of tracheoesophageal puncture (TEP) voice. To date, little information exists regarding the vibratory characteristics of the PES. The purpose of this investigation was to study the anatomy and physiology of the PES using videostroboscopy. STUDY DESIGN: Prospective study investigating the anatomy and physiology of the PES in 34 laryngectomees who used TEP speech as their primary form of communication. MATERIALS AND METHODS: Videostroboscopy and voice recordings were graded by three trained, blinded judges using a seven-point scale. RESULTS: The patients demonstrated differences that allowed for separation of patients into two main groups: "poor" and "effective" TEP speakers. The voice quality differences were explained by anatomic and physiologic characteristics of the PES. Redundant, thick, and dyssynchronous PES features were observed in patients with poor TEP speech skills; the effective speakers exhibited less redundant, thinner mucosa and more synchronous vibratory patterns. Moreover, the latter subgroup consistently demonstrated a greater degree of volitional PES control and less spasmodic activity than their poorly speaking counterparts. Length of the PES opening (measured in the horizontal plane) as well as amount and consistency of secretions did not appear to influence TEP speech or voice proficiency. CONCLUSION: Videostroboscopy in laryngectomees is a noninvasive, inexpensive, easily performed procedure that may contribute valuable information regarding the anatomy and physiology of the PES, especially in patients who experience difficulties achieving satisfactory TEP voice and speech production. PMID- 9851491 TI - Functional magnetic resonance imaging of auditory cortex in children. AB - OBJECTIVE: TO obtain images of auditory cortex activation in children by using functional magnetic resonance imaging (FMRI). METHODS: Seven healthy children (three girls and four boys), ages 6 to 10 years, were studied. Hearing evaluation was performed by puretone audiometry on the day of FMRI study. Brain imaging was performed on a commercial 1.5 T imager using a three-axis local gradient coil. During scanning the children were instructed to lie still and avoid any lip, eye, jaw, or other facial movements. Subjects were asked to listen to a standard text presented in on-off sequences. Functional images of the auditory cortex were acquired with FMRI technique. Functional imaging processing was done using cross correlation techniques with a coefficient of 0.5 (P < .0001). RESULTS: Functional correlation images of the auditory cortex activation were obtained in six of seven children after image processing. All children showed activation in the superior temporal gyrus, Heschl's gyrus, planum temporale, frontal lobe, and parietal regions. There was no significant difference in the number and percentage of activated pixels on right and left auditory cortices. CONCLUSIONS: Functional images of auditory cortex activation were obtained in healthy children following binaural text presentation. Consistent activation was observed in primary and secondary auditory cortices with no hemispheric dominance. FMRI characteristics of the auditory cortex activation in healthy children should be established in order to study those with hearing impairment. PMID- 9851492 TI - Endoscope-assisted vestibular neurectomy. AB - OBJECTIVE/HYPOTHESIS: In some instances endoscopes offer better visualization than the microscope and frequently allow less invasive surgery. This study was undertaken to determine whether endoscopy is safe and effective during neurectomy of the vestibular nerve. METHOD: Ten patients with intractable unilateral Meniere's disease underwent a retrosigmoid craniotomy for neurectomy of the vestibular nerve. Endoscopy with a Hopkins telescope was used during each procedure to study posterior fossa anatomic relationships and to assist the neurectomy. Preoperative and postoperative audiometric evaluation was performed in all patients undergoing vestibular neurectomy. Nine of these patients had preoperative electronystagmography, and four patients completed postoperative electronystagmography. The 1995 American Academy of Otolaryngology-Head and Neck Surgery's Committee on Hearing and Equilibrium guidelines for the diagnosis and evaluation of therapy in Meniere's disease were used. RESULTS: Complete neurectomy was achieved in all 10 patients. Endoscopy allowed improved identification of the nervus intermedius and the facial, cochlear, and vestibular nerves and adjacent neurovascular relationships without the need for significant retraction of the cerebellum or brainstem. In addition, endoscopic identification of the cleavage plane between the cochlear and vestibular nerves medial to or within the internal auditory canal (n = 3) was not made with the 0-degree endoscope; however, identification was made with the 30- or 70-degree endoscope in all cases. In all patients with Meniere's disease, elimination of the recurrent episodes of vertigo (n = 10) or otolithic crisis of Tumarkin (n = 1) was achieved. CONCLUSIONS: Posterior fossa endoscopy can be performed safely. Endoscope-assisted neurectomy of the vestibular nerve may offer some advantages over standard microsurgery including increased visualization, more complete neurectomy, minimal cerebellar retraction, and a lowered risk of cerebrospinal fluid leakage. PMID- 9851493 TI - A review of 308 cases of revision stapedectomy. AB - OBJECTIVE/HYPOTHESIS: Identify causes of primary and revision stapedectomy failure in 308 patients, assess whether these are different based on source of initial surgery, and evaluate hearing results in revision stapedectomy to improve outcome. STUDY DESIGN: Retrospective, nonrandomized chart review of patients undergoing revision stapedectomy in a referral otology practice in a large metropolitan region. MATERIALS AND METHODS: Intraoperative findings, preoperative and postoperative revision stapedectomy air and bone conduction pure-tone averages, speech discrimination scores, postoperative air-bone gaps, complications, and repeated revisions were noted in 308 patients. RESULTS: Leading causes of primary stapedectomy failure included dislocated prosthesis (24.4%), inadequate prosthesis length (14%), long process resorption (14%), and fibrous adhesions (13.6%). Revision stapedectomy air-bone gaps were less than 10 dB in 80% and greater than 30 dB in 6.8% of cases. Increased sensorineural hearing loss occurred in 0.8% of revision stapedectomy cases. Five of seven cases of vertigo associated with primary stapedectomy resolved after revision surgery. CONCLUSION: Revision stapedectomy by experienced surgeons is highly effective in attaining successful air-bone gap closure in 80% and improved closure in 84.8% of operative cases. Risk of vertigo and/or sensorineural hearing loss was not any higher in this patient population when compared with reports of primary stapedectomy. PMID- 9851494 TI - Congenital aural atresia: stability of surgical results. AB - OBJECTIVES/HYPOTHESIS: To compare early (<1 y) and longer-term (1.0-7.5 y; mean follow-up, 2.8 y) hearing results following surgery for congenital aural atresia and to analyze revision cases for cause of failure, time of occurrence, and outcome. STUDY DESIGN: Retrospective study of 55 consecutive patients (59 ears) undergoing surgery during an 11-year period for congenital aural atresia. METHODS: The best speech reception threshold (SRT) during the first postoperative year was compared with the most recent SRT beyond the first postoperative year. The complication rate and long-term hearing results (>1 y) for initial and revision surgeries were compared. RESULTS: In the early postoperative period, an SRT of < or =25 dB was achieved in 60% and an SRT of < or =30 dB in 70%. With longer follow-up (mean, 2.8 y), 46% of patients maintained an SRT < or =25 dB. Revision surgery was necessary in approximately one third of patients. Longer term hearing results following initial surgery only or after revision surgery showed an SRT < or =25 dB in 53% and an SRT < or =30 dB in 64%. For primary and revisions surgeries the incidence of temporary facial paralysis was 1.5% and for significant hearing loss, 3.0%. CONCLUSIONS: Some degradation in hearing does occur as patients are followed beyond the first postoperative year. Revision surgery will be necessary in approximately one third of cases. With revisions, an SRT < or =25 dB can be expected in half of cases and an SRT < or =30 dB in two thirds of cases. The ability to provide these hearing results supports this surgery in unilateral cases. PMID- 9851495 TI - Airway management in children with major craniofacial anomalies. AB - OBJECTIVES: Delineation of clinical characteristics affecting the airway in a cohort of craniofacially deformed children. What factors differ between patients requiring and those not requiring surgical airway intervention? What factors predispose to the need for tracheotomy? When can decannulation be expected if tracheotomy is required? What interventions aid decannulation? STUDY DESIGN: Five year retrospective chart review at tertiary center. METHODS: Two hundred fifty one patients met the following entry criteria: enrollment in the New York University Institute of Reconstructive and Plastic Surgery's Craniofacial Clinic and admission to Tisch Hospital in Manhattan for surgery from 1990 to 1994. Hospital, clinic, and departmental office records were reviewed. All patients had major craniofacial bony anomalies and underwent administration of general anesthesia at least once. RESULTS: Nearly 20% of all children required tracheotomy (47/251). Craniofacial synostosis patients (Crouzon, Pfeiffer, or Apert syndrome) had the highest rate of tracheotomy (48% [28/59]). Mandibulofacial dysostoses patients (Treacher Collins or Nager syndrome) had the next highest rate (41% [28/59]). Patients with oculo-auriculo-vertebral sequence were less likely to undergo tracheotomy (22% [9/41]). Children with craniosynostosis rarely required a surgical airway, unless there was marked associated facial dysmorphism (1% [1/72]). The duration of cannulation was related to the age at tracheotomy in a bimodal distribution. Generally, tracheotomies required before age 4 years remained for several years, whereas those placed after age 4 were removed after several weeks. The presence of a cleft palate correlated with reduced risk for tracheotomy, but the presence of a ventriculoperitoneal shunt correlated with an increased risk for tracheotomy. Procedures selectively used to improve the airway included midface advancement, mandibular expansion, tonsillectomy and adenoidectomy, uvulopalatopharyngoplasty, anterior tongue reduction, and endoscopic tracheal granuloma excision. CONCLUSIONS: The likelihood for surgical airway management is related to specific craniofacial diagnosis. The length of tracheal cannulation is greatest for infants and young children who manifest severe airway compromise, often because of nasal obstruction in combination with other anatomic factors. Early tracheotomy is advocated for these patients to promote optimal growth and development. Choanal atresia is often misdiagnosed in these infants; nasal obstruction is actually secondary to midface retrusion. Staged surgical interventions can allow eventual successful decannulation in nearly all cases of craniofacial syndromes. PMID- 9851496 TI - Safety of paranasal sinus surgery in patients with cystic fibrosis. AB - OBJECTIVE: Document the safety of paranasal sinus surgery in cystic fibrosis patients and review the changing trends in paranasal sinus surgery in the cystic fibrosis population. STUDY DESIGN: Retrospective review. MATERIALS AND METHODS: Chart review of cystic fibrosis patients who underwent paranasal sinus surgery from 1955 to 1997. RESULTS: Indications for surgery included chronic sinusitis, nasal obstruction, purulent rhinorrhea, head pain, and pyocele. Average duration of anesthesia was 2.1 hours. The complication rate from general anesthesia was zero. Excessive bleeding and significant hypoxia did not occur. CONCLUSIONS: Paranasal sinus surgery and general anesthesia can be safely performed in cystic fibrosis patients. The indications for paranasal sinus surgery are changing from symptomatic nasal obstruction to pre-lung transplantation care. Today, treatment has evolved to include placement of sinus catheters for direct topical instillation of antibiotics and consideration of maxillary and frontal sinus obliteration. PMID- 9851497 TI - Changes in nasal epithelium in patients with severe chronic sinusitis: a clinicopathologic and electron microscopic study. AB - OBJECTIVE: Defective ciliary ultrastructure and impaired mucociliary clearance play an important role in the development of respiratory disease and sinusitis. Changes in the ciliary ultrastructure of the sinonasal epithelium have been documented in patients with primary ciliary dyskinesia. However, secondary ciliary dyskinesias and epithelial cytopathologic changes have been underappreciated as a consequence of respiratory dysfunction and chronic sinusitis. STUDY DESIGN: Thirty-two patients with severe chronic sinusitis were evaluated for ciliary and epithelial abnormalities. MATERIALS AND METHODS: Fourteen patients (44%) were children who underwent full allergy, sweat, and immunologic workups. Eighteen patients (56%) were adults who had severe refractory sinusitis and had failed previous sinus surgery. All patients underwent nasal epithelium biopsies of the middle turbinate and evaluation by light and transmission electron microscopy. RESULTS: Ciliated cells were found in 23 patients (72%) with 9 patients (28%) having no cilia. Foci of normal ciliated epithelium were found in only 19% of the patients, often in epithelial invaginations. Variable numbers (usually a minor population) of cilia in 20 cases (87%) exhibited ultrastructural defects including compound cilia and microtubule and dynein arm defects. All of the patients showed variable loss of differentiated epithelial cells ranging from denuded epithelium to basal cell hyperplasia often associated with squamous metaplasia, secondary to chronic sinonasal disease. The lamina propria was often edematous with dilated capillaries, plasma cells, lymphocytes, and hyperplastic seromucous glands. CONCLUSIONS: This study demonstrates that ciliary dyskinesias are primarily the result rather than the cause of chronic sinusitis. Patients with chronic sinusitis of uncertain origin exhibit a prominent loss of differentiated epithelial cells, as well as ciliary defects, most of which are likely to be secondary to the chronic disease process. These changes slow down mucociliary clearance and lead to a vicious cycle leading to chronicity. PMID- 9851498 TI - Long-term follow-up after subtotal parathyroidectomy in patients with renal failure. AB - OBJECTIVES/HYPOTHESIS: The most appropriate type of surgery for hyperparathyroidism secondary to renal failure remains controversial. We report a 5-year experience of patients with hyperparathyroidism secondary to end-stage renal disease who underwent subtotal parathyroidectomy. We believe that this is the procedure of choice, offering several advantages over total parathyroidectomy with and without reimplantation. STUDY DESIGN: Retrospective review. METHODS: Review of 14 consecutive renal failure patients who underwent subtotal parathyroidectomy by one surgeon (A.K.) was performed. Follow-up ranged from 4 to 54 months. All patients were receiving chronic maintenance dialysis. All patients came to surgery with clinical symptoms of parathyroid bone disease, elevated serum calcium levels (10.1-12.4 mg/dL), and intact parathyroid hormone levels (619-4160 pg/mL), despite maximal medical therapy. At exploration four glands were identified in all patients and three and a half were removed. RESULTS: All patients experienced symptomatic relief postoperatively with normalization or near-normalization of serum calcium concentration and intact parathyroid hormone concentrations. One patient developed recurrent disease 4 months after surgery, and on re-exploration a supernumerary substernal gland was identified. A second patient developed recurrent symptoms 4 years after surgery and at the time of this writing was awaiting re-exploration. CONCLUSIONS: All patients had either resolution of or marked improvement in their subjective complaints. There have been no cases of permanent hypoparathyroidism. We believe that subtotal parathyroidectomy is the best procedure for patients with refractory symptoms of secondary hyperparathyroidism. PMID- 9851499 TI - Early postoperative calcium levels as predictors of hypocalcemia. AB - OBJECTIVE: Early, safe postoperative hospital discharge has become a priority in the current environment of cost containment and managed care. One determinant for this following operations of the thyroid or parathyroid glands is a stable postoperative calcium level. This study sought to determine whether early postoperative calcium levels could safely predict hypocalcemia following operations of the thyroid or parathyroid glands. STUDY DESIGN: A retrospective chart review of 197 patients undergoing such operations was undertaken. METHODS: The study population was divided into patients undergoing parathyroid exploration with removal of one or more glands for parathyroid disease ("parathyroid" group) and patients undergoing bilateral thyroid operations ("nonparathyroid" group). Postoperative calcium levels were plotted as a function of time, and the slope between the first two postoperative calcium levels examined. RESULTS: A positive slope predicted normocalcemia in 100% of "nonparathyroid" and 90% of "parathyroid" surgeries. A negative slope was predictive in its magnitude. The nonparathyroid patients who developed postoperative hypocalcemia had an average slope significantly more negative (-0.84% change/h) than patients remaining normocalcemic (-0.49% change/h) (P = .03). Similarly, the parathyroid patients who became hypocalcemic had an average initial slope nearly twice as negative ( 1.4% change/h) as patients remaining normocalcemic (-0.78% change/h) (P = .005). CONCLUSIONS: These results indicate that an initial upsloping postoperative calcium curve based on two early postoperative calcium measurements is strongly predictive of a stable postoperative calcium level, and a steeply downsloping initial calcium curve is worrisome for eventual hypocalcemia. PMID- 9851500 TI - Noise-induced hearing loss in young adults: the role of personal listening devices and other sources of leisure noise. AB - CONCEPT: No consensus exists regarding the magnitude of the risk of noise-induced hearing loss (NIHL) associated with leisure noise, in particular, personal listening devices in young adults. OBJECTIVE: Examine the magnitude of hearing loss associated with personal listening devices and other sources of leisure noise in causing NIHL in young adults. STUDY DESIGN: Prospective auditory testing of college student volunteers with retrospective history exposure to home stereos, personal listening devices, firearms, and other sources of recreational noise. METHODS: Subjects underwent audiologic examination consisting of estimation of pure-tone thresholds, speech reception thresholds, and word recognition at 45 dB HL. RESULTS: Fifty subjects aged 18 to 30 years were tested. All hearing thresholds of all subjects (save one-a unilateral 30 dB HL threshold at 6 kHz) were normal, (i.e., 25 dB HL or better). A 10 dB threshold elevation (notch) in either ear at 3 to 6 kHz as compared with neighboring frequencies was noted in 11 (22%) subjects and an unequivocal notch (15 dB or greater) in either ear was noted in 14 (28%) of subjects. The presence or absence of any notch (small or large) did not correlate with any single or cumulative source of noise exposure. No difference in pure-tone threshold, speech reception threshold, or speech discrimination was found among subjects when segregated by noise exposure level. CONCLUSION: The majority of young users of personal listening devices are at low risk for substantive NIHL. Interpretation of the significance of these findings in relation to noise exposure must be made with caution. NIHL is an additive process and even subtle deficits may contribute to unequivocal hearing loss with continued exposure. The low prevalence of measurable deficits in this study group may not exclude more substantive deficits in other populations with greater exposures. Continued education of young people about the risk to hearing from recreational noise exposure is warranted. PMID- 9851501 TI - Relationship between middle ear pressure, mucosal lesion, and mastoid pneumatization. AB - OBJECTIVES: The inflammatory changes of pneumatic space mucosa are thought to affect the development of pneumatic space and the function of transmucosal gas exchange. So, it is assumed that the mucosal change is deeply involved in the onset and healing process of otitis media with effusion (OME). The objective of this study is to investigate whether the growth of the mastoid cells and the middle ear transmucosal gas exchange function in patients with OME are affected by the histopathologic changes in middle ear mucosa, and whether these two factors are changed by the treatment of OME. MATERIALS AND METHODS: From 65 children with OME, middle ear mucosal specimens were collected during indwelling of a tympanic tube, and their histopathologic changes were compared with cell growth area measured on radiographic films and with the change in the middle ear total pressure (METP). RESULTS: Suppression of the cell growth area was stronger and the METP stayed lower in cases with a higher degree of mucosal lesion. Significant expansion of pneumatic area was observed after 2 years of tube indwelling, and the peak METP showed a significant increase after 1.5 years. This increase in the METP was caused by diffusion/absorption of CO2 and O2 and indicated recovery of the transmucosal gas exchange function. CONCLUSIONS: The growth of the mastoid cells and the middle ear transmucosal gas exchange function are closely affected by the subepithelial histopathologic changes in middle ear mucosa, and these two factors recover by the treatment of OME. PMID- 9851502 TI - Development of the tubotympanum in the rat. AB - OBJECTIVE: To investigate the relationship between the anatomical maturation of the middle ear and that of the eustachian tube and paratubal muscles in the rat. DESIGN: Wistar rats ranging from gestational day 12 to postnatal day 40 were used. METHODS: Tissue specimens were examined with routine light microscopy and electron microscopy. Epithelial differentiation was studied immunohistochemically with antibodies to different cytokeratins. RESULTS: The epithelial lining of the tubotympanum showed differentiation-related cytokeratin expression throughout the whole developmental period. The mucociliary epithelium reached mature features around birth. A dorsal extension and its framing cartilage started forming around 5 days after birth. This extension became lined by stratified nonciliated epithelium and attained maturity around 10 days after birth concurrently with the attachment of the dilatory muscles. This process was immediately followed by aeration of the middle ear cavity. CONCLUSIONS: The continuous expression of cytokeratins demonstrates that the epithelial lining of the tubotympanum is only derived from the embryonal endoderm. Furthermore, this study demonstrates that the eustachian tube shows a two-stage postnatal development. First, the mucociliary system matures, providing protection/clearance when the animal starts respiration and swallowing. Subsequently, the dorsal part attains maturity. The features of the epithelial lining of the dorsal part of the eustachian tube and the coincidence of the maturation of this part with the attachment of the dilating muscle fibers and the aeration of the middle ear indicates that this part provides ventilation. These findings support the authors' hypothesis that different parts of the eustachian tube serve different purposes: clearance, protection and ventilation. PMID- 9851503 TI - Second malignant neoplasia in patients with T1 glottic cancer treated with radiation. AB - OBJECTIVE/HYPOTHESIS: To evaluate incidence, site of occurrence and outcome of second malignant neoplasia (SMN) in patients with T1 glottic cancer treated with radiation. STUDY DESIGN: Retrospective. METHODS: Between February 1964 and May 1993, 158 patients with T1 squamous carcinoma of the larynx were treated with definitive radiation. Incidence, site (aerodigestive tract or not) and outcome of SMN were analyzed. Median follow-up was 63 months (range, 12-245 mo). RESULTS: Thirty-four patients developed SMN, for an overall incidence of 22%. Twenty-four (67%) SMNs occurred in an aero-upper-digestive-tract site compared with nine (25%) occurring in a non-aero-upper-digestive tract site. The incidence of SMN observed was higher than would be expected for the general population at risk. The observed-to-expected ratios (OER) for all SMN, aero-upper-digestive SMN and non-aero-upper-digestive SMNs were 1.73, 5.53, and 0.62, respectively. Overall 5- and 10-year survivals were 76% and 57%, respectively, for those who did not develop SMN, as compared with 68% and 26%, for those who developed SMN (P = .003). Overall, 13 patients (8.2%) have died from laryngeal cancer, while 23 (15%) died from SMN (P = .001). CONCLUSION: This study confirms a higher incidence of SMN in T1 glottic cancer patients, compared with the general population. The majority of cases occur in aero-upper-digestive sites. These patients are more likely to die from their SMN than from glottic cancer. Patients with T1 squamous cell carcinoma of the glottic larynx represent a group of head and neck cancer patients who should be targeted in studies evaluating the potential benefits of chemoprevention, and aggressively counseled for social and/or behavioral modifications. PMID- 9851505 TI - Prospective randomized comparison of endonasal endoscopic dacryocystorhinostomy and external dacryocystorhinostomy. AB - OBJECTIVES AND STUDY DESIGN: The advent of the rigid endonasal endoscope and the development of functional endoscopic sinus surgery (FESS) technique have awakened interest in an endonasal endoscopic dacryocystorhinostomy (EESC-DCR) in treating nasolacrimal obstruction. This prospective, randomized study compares EESC-DCR with traditional external dacryocystorhinostomy (EXT-DCR) for their success rates, surgical duration, and postoperative symptoms. PATIENTS AND METHODS: Sixty four cases in 60 patients with primary acquired nasolacrimal sac or duct obstruction were divided into two subgroups by symptoms (simple epiphora/chronic dacryocystitis). These patients were randomized within both subgroups into two operation groups. Altogether 32 EESC-DCRs and 32 EXT-DCRs were performed. The final follow-up visit was at 1 year. The patency of the lacrimal passage was investigated by irrigation and patients were questioned about their symptoms. RESULTS: The success rate at 1 year after surgery was 75% for EESC-DCR and 91% for EXT-DCR after primary surgery. The difference was not statistically significant (P = .18). The success rate after secondary surgery with a follow-up time of 1 year was 97% in both study groups. The average duration for EESC-DCR was 38 minutes, and 78 minutes for EXT-DCR, (P < .001). CONCLUSIONS: EXT-DCR, when compared with EESC-DCR, appears to give a higher, although not statistically significant, primary success rate, but the secondary success rates are equal, indicating that these two different DCR techniques are acceptable alternatives. PMID- 9851504 TI - Pretreatment and midtreatment measurement of oxygen tension levels in head and neck cancers. AB - OBJECTIVE: Considerable evidence exists to suggest that tumor hypoxia results in radioresistance. Historically, it has been difficult to assess tumor oxygen tension levels reliably. These levels can now be assessed in head and neck malignancies using the Eppendorf pO2 histograph, which uses a fine-needle electrode and a computerized micromanipulator. This technology was used to compare the pretreatment tumor oxygen tension level in lymph node metastases of patients with head and neck cancer to measurements taken during nonsurgical treatment after a partial response had been achieved. STUDY DESIGN: Prospective study. METHODS: Oxygen tension levels were measured in the cervical lymph nodes of 10 patients with biopsy-proven head and neck squamous cell carcinoma and cervical metastases who were being treated with nonsurgical management. These levels were obtained using the Eppendorf pO2 histograph system. Measurements were taken before the start of treatment and were repeated when the size of the cervical metastatic node had decreased by 50%. Normal subcutaneous tissue was measured during the same session. The mean and median pO2 levels, as well as the percentage of measurements with pO2 less than 5 mm Hg were determined. RESULTS: A mean of 72.6 measurements per session was taken from each lymph node. The median tumor pO2 measurement fell from a mean (+/-SD) of 13.9+/-8.0 mm Hg to 7.3+/-9.9 mm Hg. Even more dramatic, however, was the substantial increase in the percentage of values less than 5 mm Hg, a rise from 29% to 52%. CONCLUSIONS: While there is variability both in the pretreatment oxygenation of head and neck cervical metastases and in the change in tumor oxygen tension during treatment, there appears to be a decrease in the overall oxygenation of the tumors. The dramatic increase in very low oxygen measurements may reflect selective survival of radioresistant or chemoresistant hypoxic tumor cells. Cells at the very low level would be expected to be radiobiologically hypoxic (resistant to radiation induced cell kill). PMID- 9851506 TI - A review of the toxicosis and biological properties of the genus Eupatorium. AB - Eupatorium genus grows wild in many parts of the world. A number of species of Eupatorium are toxic to grazing animals. Milk sickness in humans is caused by ingestion of milk of the animals reared on the pastures infested with Eupatorium rugosum (white snakeroot). While some information is available on the toxins in various species of Eupatorium, ambiguities still persist in extrapolation of the data to field incidence of toxicosis. Eupatorium genus has been used for its medicinal properties for many decades. A number of bioactive natural products have been reported in the extracts of Eupatorium spp. and the genus is a promising bioresource for preparation of drugs and value-added products. PMID- 9851507 TI - Haemorrhagic activity of neotropical rattlesnake (Crotalus vegrandis Klauber, 1941) venom. AB - In this work we have demonstrated for the first time in any Venezuelan Crotalus, haemorrhagic activities that are present in the Neotropical Uracoan rattlesnake (Crotalus vegrandis) venom. This venom has been little studied, perhaps because the snake is restricted to a small habitat located in the dry savannahs of northeastern Venezuela. In our experiments Crotalus vegrandis venom caused a very evident haemorrhagic area consisting of approximately 2/3 diameter of the area caused by a positive control Bothrops lanceolatus venom. Crotalus vegrandis venom affects blood coagulation and causes intense haemorrhages. It does not clot fibrinogen, therefore it has neither thrombin-like activity which transforms fibrinogen to fibrin nor procoagulant enzymatic function which produces thrombin. On the other hand, it degrades fibrinogen making it incoagulable to thrombin. The venom, when injected in the animals, resulted in a high increase of the Partial Time of Thromboplastin (PTT) tests. It is interesting to observe that the haemorrhagic capacity in the Crotalus genus (widely distributed in the American continent) increases from south to the north, being present in North American Crotalus, a venom with wide haemorrhagic activities, and almost non-existent in most of the South American Crotalus species and subspecies. PMID- 9851508 TI - Differential biodistribution of native and 2 kGy 60Co irradiated crotoxin in tissues of CBA/J mice. AB - Crotalus durissus envenomation is treated using antivenins produced in horses. During production, animals have problems, sometimes followed by death, due to the high toxicity of the main toxin, crotoxin. Several methods tested to detoxify this toxin often resulted in decreased immunogenicity. Gamma irradiation has proved to be a successful method for crotoxin detoxification without loss of immunogenicity. We have studied the biodistribution of 2 kGy 60Co irradiated crotoxin (iCTX) in mouse tissues. We used both 125I-labeled iCTX or its detection by a specific immunohistochemistry assay (IHA). Both approaches showed similar early excretion of toxins by the kidneys. Higher iCTX uptake was seen in spleen and liver, which are rich in immune responder cells. In contrast to previous reports concerning native crotoxin (nCTX), we failed to detect iCTX in the neuromuscular junction, but both toxins were found on the kidney tubular cell surface, with rapid excretion that was more intense for iCTX. Kupffer cells and splenocyte macrophages presented IHA staining, as shown by the increased uptake of 125I toxin by these organs. No staining was observed in the brain, lung or heart, which also showed very low 125I counts. Allied to reduced toxicity, irradiation induced early endocytosis of crotoxin by phagocytic cells, improving antigen processing. PMID- 9851509 TI - Effects of Stachybotrys chartarum (atra) conidia and isolated toxin on lung surfactant production and homeostasis. AB - This study evaluated the effects of Stachybotrys chartarum conidia and a trichothecene, isosatratoxin-F, on choline incorporation into DSPC by fetal rabbit alveolar type II cells and on alveolar surfactant subtypes in mice. Exposure of fetal rabbit type II cells to S. chartarum conidia at concentrations of 10(3) to 10(6) conidia ml(-1) significantly depressed [3H] choline incorporation after 24 h of exposure. Exposure of the rabbit cells to 10(5) to 10(6) conidia ml(-1) also resulted in significantly depressed [3H] choline uptake after 48 h. Additionally, fetal rabbit alveolar type II cells exposed to isosatratoxin-F in concentrations ranging from 10(-9) to 10(-4) M showed a significant reduction in [3H] choline incorporation into DSPC. Alveolar surfactant phospholipid concentrations in the different metabolic subfractions of lung lavage fluid of mice intratracheally exposed to either 50 microl of 10(7) ml(-1) S. chartarum conidia or 50 microl 10(-7) M isosatratoxin-F showed some significant changes at 12, 24, 48, and 72 h post-exposure, compared to the surfactant subfractions of control mice which were either untreated, exposed to saline or to 50 microl of 10(-7) ml(-1) Cladosporium cladosporioides conidia. In both the S. chartarum- and the isosatratoxin-F-treated mice, exposure significantly increased P10, P100, and S100 phospholipid concentrations, while the P60 phospholipid concentrations were depressed. In contrast, C. cladosporioides-treated mice showed only one significant change in subfraction phospholipid concentration: P60 was depressed at 48 h post-exposure. These results reveal that alveolar type II cells are sensitive to exposure to S. chartarum conidia and to isosatratoxin F. Sensitivity is manifest by alterations in the normal metabolic processing of alveolar surfactant. In exposed mice, this effect appears to involve a significant increase in newly secreted surfactant and an accumulation of the used surfactant forms. PMID- 9851510 TI - Biological activities of prymnesin-2 isolated from a red tide alga Prymnesium parvum. AB - Previously we isolated prymnesin-1 (PRM1) and prymnesin-2 (PRM2) as the major hemolytic and ichthyotoxic agents in the red tide organism Prymnesium parvum and disclosed the structure of PRM2 as a novel glycoside with unusual multiple functionality. PRM2 caused 50% hemolysis of a 1% suspension of dog red blood cells at 0.5 nM. The potency exceeded that of plant saponin by 50000 times. The lethality of PRM2 on freshwater fish Tanichthys albonubes was comparable to that of brevetoxin and also the ichthyotoxicity was markedly enhanced by Ca2+ and by a slight elevation of pH: LC50 in a Ca2+ free medium (pH 7.0) was 300 nM and in the presence of 2 mM Ca2+ (pH 8.0) was 3 nM. The hemolytic activity of PRM2 was not affected by Ca2+ but was markedly affected by blood cell origin. Also, the observation of competitive inhibition by the PRM2 analogues allowed us to assume the presence of a specific binding site on the blood cell surface. PMID- 9851511 TI - Placental and lactational transfer of ochratoxin A in rats. AB - The placental and lactational transfer of ochratoxin A (OA) was investigated in a cross-fostering study in rats. Dams were given 50 microg OA(-1) kg body weight by gastric intubations 5 times a week for 2 weeks before mating, during gestation and then 7 days a week during lactation. Neonates from OA-treated dams were cross fostered at birth to control dams treated with only vehicle. In the same way, neonates from control dams were cross-fostered to OA-treated dams. Treatment with OA did not result in any effects on birth weight or growth development of the pups during the first 21 days of life. There were no effects on milk quality as measured by milk lipids, protein or lactose concentrations, or on milk production, assessed by the mammary gland content of RNA and DNA. A mean milk:blood ratio of approximately 0.6 was found. The dose of OA from milk to the suckling pup at 14 days of age can be calculated to about 50 microg kg(-1) body weight(-1) day, which is similar to the dose given to the dams. Pups exposed to OA only via milk had blood and kidney levels of OA approximately 3 times higher than their dams, indicating a high absorption and/or a low excretion of OA in the sucklings. At 14 days of age the highest blood and kidney levels of OA were found in offspring exposed both via placenta and milk, with the highest contribution from milk. Offspring exposed only via milk had about 4-5 times higher levels of OA in blood and kidney compared to offspring exposed only via placenta. As milk could be a significant source of OA exposure in newborns, adverse health effects resulting from postnatal exposure should be studied and evaluated in the risk assessment of OA. PMID- 9851512 TI - From antigen presentation to T-cell activation. PMID- 9851513 TI - Development of accessory phenotype and function during the differentiation of monocyte-derived dendritic cells. AB - Dendritic cells (DCs) are professional antigen-presenting cells (APCs), and classical DCs, such as Langerhans cells (LCs) or interdigitating DCs (IDCs) are known to be the most potent stimulators of T lymphocytes. Earlier, several groups described the generation of DCs from monocytes, starting with peripheral blood mononuclear cells (PBMCs), adherent cells or magnetic bead-purified CD14+ cells. Although modifications of the original protocols have already been described, some questions relevant to clinical application and basic studies have not yet been addressed. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL4) appear to be necessary, but are not sufficient for the differentiation of monocyte-derived dendritic cells (MoDCs), as indicated by the failure to generate such cells under serum-free conditions. Using adherence purified monocytes, we first investigated the amount of GM-CSF and IL4 required for the differentiation of DCs. Consecutive kinetic studies during the differentiation period were designed to demonstrate how monocytes acquire the phenotype and function of DCs. The results showed that small amounts of GM-CSF and IL4 were required to generate MoDC which acquired their phenotype and function within 4 days. IL13 may substitute for IL4, whereas IL10, TNF alpha or IFN gamma inhibited the generation of MoDCs. PMID- 9851514 TI - Langerhans cells do not express alternative macrophage activation-associated CC chemokine (AMAC)-1. AB - We have cloned a novel human CC chemokine, alternative macrophage activation associated CC chemokine (AMAC)-1 that is highly homologous to macrophage inflammatory protein (MIP)-1alpha. In contrast to MIP-1alpha, AMAC-1 is induced in macrophages by Th2-associated cytokines IL4, IL13, and IL10 in vitro; in addition, AMAC-1 is expressed by Th1-suppressive alveolar macrophages in vivo. Surprisingly, however, AMAC-1 is also expressed by GM-CSF-induced, in vitro monocyte-derived dendritic cells when treated by IL4. Here, we present a detailed analysis of AMAC-1 expression in monocyte-derived dendritic cells in vitro and show that the prime dendritic cells in vivo, i.e. epidermal Langerhans cells, do not express AMAC-1 mRNA. In conclusion, AMAC-1 is a novel CC chemokine whose Th2 associated expression pattern in alternatively activated suppressor macrophages in vivo and in vitro and its absence from epidermal Langerhans cells in vivo suggest that it may be involved in inhibition of Th1 reactions and in tolerance induction. PMID- 9851515 TI - Regulation of chemokine receptor expression in dendritic cells. PMID- 9851516 TI - Monocyte-derived dendritic cells: development of a cellular processor for clinical applications. AB - Since dendritic cells (DCs) are the most professional antigen-presenting cells, (Schuler et al., 1997), increasing interest in their use in clinical approaches has been observed. (Nestle et al., 1998; Murphy G. et al., 1996). We have developed an ex vivo standardized process for the generation of dendritic-like cells (MAC-DCs) from human blood circulating monocytes. Human monocytes can differentiate into very different functional cells according to the conditions of culture, media and cytokines used. In the present study, we demonstrate that both pure monocytes and mononuclear cells differentiate into DCs when they are grown in defined medium AIM-V in the presence of granulocyte-macrophage colony stimulating factor (GM-CSF) plus IL13 and in approved biocompatible non-adherent bags. Quality and functional controls of the immature DCs obtained rely on bacterial sterility, viability, morphology and recovery. The MAC-DCs also present an immature DC phenotype with a low expression of CD14 and CD64, and high expression of MHC-I, MHC-II and CD40. They also express B7 costimulatory molecules (CD80, CD86), CD83, and CD1a molecules. They induce strong allogenic T cell proliferation (mixed lymphocyte reaction as well as proliferation of autologous memory T lymphocytes when incubated in the presence of recall antigens (tuberculosis, Candida albicans, and tetanus toxoid). They also show an increase in phagocytic uptake of yeast, tumour cells and debris. The global closed system which, under reproducible good medical practice (GMP) conditions, enables the production of dendritic cells of clinical quality, has been optimized ("Vac Cell Processor"). It contains all bags, connections, media, reagents, washing solutions, control antibodies, standard operating procedures, data management, traceability and help in the form of dedicated software. PMID- 9851517 TI - Cellular vaccines. AB - This project is devoted to the development of novel cellular vaccines designed to treat cancer patients. These cellular vaccines present and enhance immunogens, which will elicit a potent immune response. The goal is to achieve safe and effective immune reaction against the patient's own tumour. (1) Autologous cellular vaccines are prepared by processing circulating blood mononuclear cells outside of the patient's body (ex vivo) to differentiate them into antigen presenting cells (APCs). Monocyte-derived APCs (MD-APCs) are then grown in the presence of exogenous target antigens (tumour cell debris, or apoptotic bodies) to become fully mature APCs. (2) Functionality for antigen presentation to T cells of ex vivo MD-APCs is evaluated in vivo. (3) Cellular vaccines are tested in selected rodent animal models. Efficiency and immune response are monitored in pertinent experimental systems for cancer. Pharmacological data are generated for clinical investigation. Tolerance and biologic effects are documented in primates. (4) The first clinical trials on cancer patients are taking place in 1998 on melanoma and prostate cancer to validate the concept. Specialized cell processors with dedicated software and standardized controls are being developed and used for the preparation of cellular vaccines. (5) The evaluation of new non viral vectors and the validation of new non-viral transfection methods of mononuclear cells with marker genes is in progress and will lead to the ex vivo transfection of genes coding for immunostimulating cytokines or for tumour antigens in MD-APCs. Efficiency will be validated in vitro and in animal models. The ex vivo and animal model studies validate the clinical relevance of this new cellular immunotechnology. Clinical validation of individual autologous cellular vaccines in specific indications for which no treatment is presently available will allow the development of cellular and gene immunotherapy for other types of cancers. PMID- 9851518 TI - Macrophage- and lymphocyte-produced Th1 and Th2 cytokines in the tumour microenvironment. PMID- 9851519 TI - Macrophage-targeted killing and vaccines. PMID- 9851520 TI - Anti-tumour immunotherapy using dendritic-cell-derived exosomes. PMID- 9851521 TI - Autologous human macrophages and anti-tumour cell therapy. AB - Most technical problems concerning the production of human macrophages have been resolved by cultures in hydrophobic plastic, gas-permeable bags. This process enables collection of non-adherent macrophages and is well adapted to the safety requirements of cell therapy. Under optimized culture conditions, about one billion macrophages are currently obtained from a single leukapheresis product. In most clinical trials, macrophages have been activated by interferon-gamma (IFNgamma). The injections have little or no toxic effect. The anti-tumour activity of the intravenous (i.v.) administrations is more pronounced in certain protocols than in others. The mechanism remains poorly understood. In vitro, the cytolytic effect of macrophages requires cell-to-cell contact but macrophages injected i.v. show no particular tropism for tumour tissue. This could result from modifications in adhesion molecules occurring during monocyte-macrophage differentiation which might modify recruitment in inflammatory foci. Macrophages can, however, infiltrate tumour cell clusters, which could explain their improved efficacy when injected intratumorally (i.t.). Moreover, several arguments would favour the use of macrophages as human tumour antigen-presenting cells (APCs). In vitro, macrophages are as efficient as monocyte-derived dendritic cells (MDDCs) in stimulating cytotoxic T lymphocyte (CTL) clones or circulating CTL precursors. PMID- 9851522 TI - Induction of antigen presentation by macrophages after phagocytosis of tumour apoptotic cells. AB - Due to their resistance to classical chemotherapies, most human colorectal cancers have a high incidence and a poor prognosis. Immunotherapy using interleukin 2 (IL2) has provided disappointing results in the treatment of these cancers. Recently, however, we have demonstrated that a treatment combining a cell-differentiating agent, sodium butyrate (NaBut) with IL2 resulted in a remission of established peritoneal colorectal carcinomatosis in rats. Separately, neither NaBut nor IL2 treatment cured these tumour-bearing rats. NaBut is known to induce cell differentiation and subsequent apoptosis in epithelial cells, while IL2 stimulates the immune cells capable of participating in tumour rejection. We postulated that the significant therapeutic effect of NaBut/IL2 treatment could be attributed to a NaBut-induced increase in the immunogenicity of the cancer cells. We report here that NaBut induced an apoptotic process in rat colon tumour cells in vivo and in vitro. We observed, in an efficient cure, colocalization of apoptotic bodies and monocytes/macrophages at the periphery of the tumour. We propose that these apoptotic bodies are phagocytosed in vivo by the macrophages. We also showed in vitro that a subpopulation of macrophages involved in the phagocytic clearance of apoptotic cells expresses cell surface molecules associated with antigen presentation and stimulates the proliferation of naive splenocytes. Our data suggest that therapies that recruit massive induction of the apoptotic process in tumour cells could favour tumour antigen presentation via their specific phagocytosis by antigen-presenting cells (APCs). We propose that the development of specific therapies that stimulate both tumour cell apoptosis and the immune system could offer new opportunities in anti-cancer treatments of poorly immunogenic cancer cells. PMID- 9851523 TI - Cancer immunotherapy by direct in vivo transfer of immunomodulatory genes. PMID- 9851524 TI - The role of the macrophage in immune regulation. AB - The macrophage plays an important role in both the innate and acquired (humoral and cellular) immune responses. Their specialized derivatives, the dendritic cells (DCs), are uniquely potent in induction of naive T and B lymphocytes, whereas macrophages influence a range of immune responses by antigen recognition, capture, clearance and transport. They recruit haemopoietic cells to local sites of inflammation and immunity and regulate their activities. We have used various myeloid-restricted membrane antigens and receptors as markers and functional contributors to these activities, and briefly review their role in immune regulation in vivo and in vitro. PMID- 9851525 TI - Survival strategies of Leishmania donovani in mammalian host macrophages. AB - Microbes have evolved a variety of strategies to survive inside their host cells. Some have the molecular machinery to survive in the hostile environment of phagolysosomes; others escape the phagosome to the more cozy environment of the cell cytoplasm; others inhibit the phagosome fusion with hydrolase-enriched endocytic organelles. This is the case for the promastigote form of the protozoan parasite Leishmania donovani which resides in a phagosome displaying poor fusogenic properties towards endosomes and lysosomes. Recent results indicate that the lipophosphoglycan (LPG), the major cell surface molecule of Leishmania, is involved in the inhibition of phagosome maturation. Further studies in our laboratories are addressing the molecular mechanisms of action of LPG to modulate phagosome fusion properties and its effect on the biogenesis of phagolysosomes. PMID- 9851527 TI - Modulation of phagosome processing as a key strategy for Mycobacterium avium survival within macrophages. PMID- 9851526 TI - Checkpoints in antibacterial immunity. PMID- 9851529 TI - Prions, new molecular forms of biological information. PMID- 9851528 TI - An endogenous human immunostimulant acting on mononuclear phagocytes: a study in laboratory mice. PMID- 9851530 TI - Human monocytes and cytokines. PMID- 9851531 TI - The contribution of inflammation to acute and chronic neurodegeneration. PMID- 9851532 TI - Macrophages and heat shock proteins. PMID- 9851533 TI - The role of cytokines in monocyte apoptosis. PMID- 9851534 TI - The interleukin-11/receptor complex: rational design of agonists/antagonists and immunoassays potentially useful in human therapy. PMID- 9851535 TI - Nitric oxide synthase inhibition by haem oxygenase decreases macrophage nitric oxide-dependent cytotoxicity: a negative feedback mechanism for the regulation of nitric oxide production. AB - Nitric oxide (NO) production in macrophages by inducible nitric oxide synthase (NOS2) has multiple tissue damaging effects and is involved in the pathogenesis of inflammation and graft rejection. Haem oxygenase (HmOx) is the enzyme which degrades haem. Its inducible isoform, HmOx1, was recently shown to increase cellular resistance against oxidative stress and to decrease inflammation and graft rejection. Since haem is an essential cofactor for NOS2 activity, we investigated the effects of HmOx1-induction upon NO secretion in macrophages. We induced HmOx1 in BALB/c bone-marrow-derived macrophages by short-term exposure to haemin (20 micromol/l, 30 min); then we incubated them for 24 h to allow maximal expression of HmOx1 activity. Next, we activated the macrophages with lipopolysaccharide (LPS) and measured their NO production and their NO-dependent cytotoxicity against P815 cells. We found that HmOx induction 24 h before LPS activation in mouse macrophages suppresses their production of NO, while HmOx inhibition (with zinc protoporphyrin) increases NO secretion. NOS2 inhibition is reflected by the decrease of macrophage NO-dependent cytotoxicity against the P815 targets. We therefore propose that HmOx1 is a physiological inhibitor of NOS2 in activated macrophages because it decreases haem availability for NOS2 synthesis. NOS2 inhibition may explain the antinflammatory effects of HmOx induction which could also be used therapeutically in situations when NO hyperproduction leads to cytotoxic effects such as inflammation or transplant rejection. PMID- 9851536 TI - Subtypes of functional alpha1- and alpha2-adrenoceptors. AB - In this review, subtypes of functional alpha1- and alpha2-adrenoceptors are discussed. These are cell membrane receptors, belonging to the seven transmembrane spanning G-protein-linked family of receptors, which respond to the physiological agonists noradrenaline and adrenaline. Alpha1-adrenoceptors can be divided into alpha1A-, alpha1B- and alpha1D-adrenoceptors, all of which mediate contractile responses involving Gq/11 and inositol phosphate turnover. A 4th alpha1-adrenoceptor, the alpha1L-, has been postulated to mediate contractions in some tissues, but its relationship to cloned receptors remains to be established. Alpha2-adrenoceptors can be divided into alpha2A-, alpha2B- and alpha2C adrenoceptors, all of which mediate contractile responses. Prejunctional inhibitory alpha2-adrenoceptors are predominantly of the alpha2A-adrenoceptor subtype (the alpha2D-adrenoceptor is a species orthologue), although alpha2C adrenoceptors may also occur prejunctionally. Although alpha2-adrenoceptors are linked to inhibition of adenylate cyclase, this may not be the primary signal in causing smooth muscle contraction; likewise, prejunctional inhibitory actions probably involve restriction of Ca2+ entry or opening of K+ channels. Receptor knock-out mice are beginning to refine our knowledge of the functions of alpha adrenoceptor subtypes. PMID- 9851537 TI - Downregulation of brain mineralocorticoid and glucocorticoid receptor by antisense oligodeoxynucleotide treatment fails to alter spatial navigation in rats. AB - Adult male Brown Norway rats were long-term intracerebroventricularly (i.c.v.) infused with antisense oligodeoxynucleotides (18-mer, double endcapped phosphorothioate protected) targeting either mineralocorticoid or glucocorticoid receptor mRNA, or received the respective mixed bases sequence or vehicle. Mineralocorticoid receptor-mixed bases and glucocorticoid receptor-mixed bases oligodeoxynucleotide infusion (1 microg/0.5 microl/h) over a time period of seven days did not alter hippocampal mineralocorticoid receptor and glucocorticoid receptor binding when compared to vehicle treatment. In contrast, i.c.v. administration of mineralocorticoid receptor, as well as glucocorticoid receptor antisense over the same time period resulted in a significantly reduced binding of mineralocorticoid receptor and glucocorticoid receptor in the hippocampus [mineralocorticoid receptor-antisense group approx. 72% of mineralocorticoid receptor-mixed bases and vehicle groups (100%); glucocorticoid receptor antisense group approx. 77% of glucocorticoid receptor-mixed bases and vehicle]. The specificity of these antisense effects is indicated by the finding that rats treated with mineralocorticoid receptor-antisense did not show any changes in glucocorticoid receptor and vice versa. Animals treated according to this infusion protocol and tested in the Morris water maze for their spatial navigation abilities failed to show significant differences among the groups. These data indicate that a reduction of hippocampal mineralocorticoid receptor or glucocorticoid receptor binding capacity by 20-30% does not interfere with spatial navigation. PMID- 9851538 TI - Antipsychotic drugs induce similar effects on the release of dopamine and noradrenaline in the medial prefrontal cortex of the rat brain. AB - In the present study we have compared the effects of the classical antipsychotic drug haloperidol and four different atypical antipsychotics (clozapine, risperidone, olanzapine, ziprasidone) on extracellular levels of dopamine and noradrenaline in the medial prefrontal cortex (MPFC) of conscious rats. Haloperidol (10, 100 and 800 nmol/kg), clozapine (0.3, 1, 10 and 30 micromol/kg), risperidone (100, 500 and 5000 nmol/kg), olanzapine (10, 100 and 500 nmol/kg) and ziprasidone (10, 100 and 1000 nmol/kg) were administered subcutaneously to rats. All compounds induced increases in dialysate levels of dopamine and noradrenaline in the medial prefrontal cortex. The increases induced by the four antipsychotic agents in extracellular levels of dopamine and noradrenaline displayed a striking co-variation both in dose and time. A similar co-variation was seen in the decrease of dopamine and noradrenaline, after administration of a low dose (30 nmol/kg, s.c.) of the dopamine D2/3 receptor agonist (+)-7-hydroxy-2-(N,N-di-n propylamino) tetralin ((+)-7-OH-DPAT). It is concluded that there is a close coupling between the release of dopamine and noradrenaline in the medial prefrontal cortex. The mechanism of action of this interaction, that might be of importance for a better understanding of the mechanism of action of antipsychotic drugs, is discussed. PMID- 9851539 TI - Characterisation of beta2-adrenoceptors, using the agonist [11C]formoterol and positron emission tomography. AB - The agonist radioligand N-[2-hydroxy-5-[1-hydroxy-2-[[2-(4-[11C]-methoxyphenyl)-1 methylethyl]am ino]ethyl]phenyl]formamide ([11C]formoterol) was synthesised in order to test its ability to visualise pulmonary beta2-adrenoceptors in vivo, with positron emission tomography (PET). Formoterol was labelled via reaction of a dibenzyl-protected precursor with [11C]CH3I. Subsequent deprotection with Pd/C and H2 yielded [11C]formoterol in 5-15% (corrected for decay) and the specific activity ranged from 5.5-22.2 TBq mmol (150-600 Ci mmol(-1)), 60-70 min after end of bombardment. Biodistribution studies with [11C]formoterol were performed in male Wistar rats which were either untreated or predosed with (D,L)-propranolol hydrochloride (2.5 mg kg(-1), beta-adrenoceptor antagonist), erythro-DL-1-(7 methylindan-4-yloxy)-3-isopropylaminobuta n-2-ol hydrochloride (ICI 118551, 0.15 mg kg(-1), beta2-adrenoceptor antagonist), isoprenaline (15 mg kg(-1), non subtype selective beta-adrenoceptor agonist) or (+/-)-(2-hydroxy-5-[2-((2-hydroxy 3-(4-((1-methyl-4-trifluoromethyl)1H-i midazol-2-yl )phenoxy)propyl)amino)ethoxy]benzamide)monomethane sulfonate (CGP 20712A, 0.15 mg kg(-1), beta1-adrenoceptor antagonist). Lungs, heart, liver and plasma were analysed for radioactive metabolites. The kinetics of [11C]formoterol in the lungs of male Wistar rats were investigated by means of a dynamic PET study. The biodistribution studies showed significant specific binding in tissues known to contain beta2-adrenoceptors (lungs, spleen, and heart). Binding in these organs was blocked by ICI 118551 and isoprenaline, but not by CGP 20712A. [11C]Formoterol was rapidly metabolised in rats but lungs and heart did not substantially take up the labelled metabolites. The binding of [11C]formoterol in various tissues of rats is consistent with the beta2-selectivity of formoterol. Whether [11C]formoterol selectively binds to the high affinity state of beta2 adrenoceptors remains to be elucidated. [11C]Formoterol is potentially useful for studying beta2-adrenoceptors with PET and this radioligand may provide new insights in the mechanisms underlying prolonged sympathomimetic action. PMID- 9851540 TI - Hydralazine-induced vasodilation involves opening of high conductance Ca2+ activated K+ channels. AB - The purpose of this study was to investigate whether high conductance Ca2+ activated K+ channels (BK(Ca)) are mediating the vasodilator action of hydralazine. In isolated porcine coronary arteries, hydralazine (1-300 microM), like the K+ channel opener levcromakalim, preferentially relaxed contractions induced by K+ (20 mM) compared with K+ (80 mM). In addition, concentration relaxation curves for hydralazine (pD2 = 5.38 +/- 0.06; Emax = 85.9 +/- 3.6%) were shifted 10-fold to the right by the BK(Ca) blockers tetraethylammonium (1 mM) and iberiotoxin (0.1 microM). In contrast, nimodipine (a Ca2+-entry blocker), relaxed contractions induced by K+ (20 mM) and K+ (80 mM) equally and nimodipine induced relaxations were neither antagonized by tetraethylammonium nor by iberiotoxin. In isolated perfused rat hearts, hydralazine (1 microM) increased coronary flow by 28.8 +/- 2.7%. Iberiotoxin (0.1 microM) suppressed this response by 82% (P < 0.05). In conscious, chronically catheterized rats the hypotensive response to hydralazine (0.6 mg kg(-1) min(-1)) was significantly reduced by 41% during infusion of iberiotoxin (0.1 mg kg(-1)). It is concluded, that opening of BK(Ca) takes part in the mechanism whereby hydralazine produces vasodilation. PMID- 9851541 TI - Prevention of arterial structural alterations with verapamil and trandolapril and consequences for mechanical properties in spontaneously hypertensive rats. AB - We compared the chronic effects in spontaneously hypertensive rats (SHR) of low doses of an angiotensin converting enzyme inhibitor, trandolapril, a Ca2+ channel antagonist, verapamil, and their combination (trandolapril-verapamil), on arterial mechanical properties, arterial wall hypertrophy and extracellular matrix proteins. Four-week-old SHR were randomly allocated to oral treatment with verapamil (50 mg kg(-1) day(-1)), trandolapril (0.3 mg kg(-1) day(-1)), the combination of verapamil (50 mg kg(-1) day(-1)) plus trandolapril (0.3 mg kg(-1) day(-1)), or placebo for 4 months. A group of Wistar Kyoto (WKY) control rats received placebo for the same period of time. At the end of the treatment, mean blood pressure was lower in verapamil-trandolapril than in trandolapril SHR, but remained higher than in WKY. Verapamil had no effects on blood pressure. Equivalent reduction in aortic wall hypertrophy was obtained in all treated SHR. Trandolapril and verapamil-trandolapril combination produced a significant reduction of aortic collagen density compared with placebo SHR. Carotid total fibronectin, EIIIA fibronectin isoform and alpha5beta1 integrin, were higher in the media of placebo SHR than in WKY. EIIIA fibronectin isoform and alpha5beta1 integrin were reduced in verapamil-SHR compared with placebo-SHR and normalized in trandolapril and verapamil-trandolapril-SHR compared with WKY. SHR-placebo and SHR treated with either verapamil or trandolapril as single-drug treatment showed a 4-fold increase in total fibronectin compared to the WKY. Only SHR treated with verapamil-trandolapril combination had total fibronectin not significantly different from that of WKY. Carotid arterial distensibility increased only in verapamil-trandolapril treated rats. Multivariate analysis showed arterial distensibility to be negatively correlated to mean blood pressure (P < 0.0001) and total fibronectin (P < 0.01). In conclusion, chronic treatment with the verapamil-trandolapril combination significantly improved in vivo arterial distensibility in SHR. The most important effects of the combination on arterial mechanics compared to those of verapamil or trandolapril alone may have been the consequence of its stronger action on arterial pressure, arterial wall hypertrophy and total fibronectin density. However we suggest that, in addition to the structural effects, complete normalization of blood pressure is necessary to obtain normal arterial distensibility. PMID- 9851543 TI - Effects of gender on the sensitivity of rat cardiac muscle to extracellular Ca2+. AB - Experiments were designed to determine if the inotropic response to increasing buffer calcium concentration differs in male and female cardiac muscle. Left atrial and papillary muscles were isolated from hearts of 3-4-month old male and female rats, bathed in Krebs-Henseleit solution (30 degrees C), and stimulated at 1.5 Hz. Isometric developed tension was monitored continuously as extracellular Ca2+ was increased in a cumulative fashion. When compared to male atrial muscle, female atrial preparations were more sensitive to the resulting positive inotropic action; EC50 values were 2.89 +/- 0.22 and 1.86 +/- 0.21 mM in male and female atria, respectively. Two-way analysis of variance (ANOVA) also indicated that there was a significant gender-associated difference in the Ca2+ dose response curves in atrial muscle. In contrast, papillary muscle did not show a significant gender-related difference in EC50 values (0.88 +/- 0.07 and 0.74 +/- 0.06 mM in males and females); however, the Ca2+ dose-response curves obtained from male and female preparations were found to be significantly different when compared by ANOVA. PMID- 9851542 TI - Pressor and bradycardic effects of tacrine and other acetylcholinesterase inhibitors in the rat. AB - The cardiovascular effects of three different acetylcholinesterase inhibitors: physostigmine, tacrine and rivastigmine injected by intravenous (i.v.) route were compared in freely moving Wistar rats. The three drugs significantly increased both systolic and diastolic blood pressure and decreased heart rate. Compared to physostigmine, a 20-fold higher dose of tacrine and a 40-fold higher dose of rivastigmine was necessary to induce a comparable pressor effect. Tacrine was chosen as a model to study the mechanisms underlying the cardiovascular effects of i.v. cholinesterase inhibitors. Atropine totally abolished while methylatropine did not affect tacrine pressor effects. Conversely, both drugs abolished tacrine-induced bradycardia. The alpha1-adrenoceptor antagonist prazosin or the vasopressin V1 receptor antagonist, [beta-mercapto-beta,beta cyclopenta-methylenepropionyl1, O-Me-Tyr2, Arg8] vasopressin partially but significantly reduced tacrine pressor effect and mostly abolished it when administered concomitantly. The tacrine pressor response was inhibited in a dose dependent manner by the i.c.v. administration of the non-selective muscarinic receptor antagonist atropine (ID50 = 1.45 microg), the muscarinic M1 receptor antagonist pirenzepine (ID50 = 4.33 microg), the muscarinic M2 receptor antagonist methoctramine (ID50 = 1.39 microg) and the muscarinic M3 receptor antagonist para-fluoro-hexahydro-sila-difenidol (ID50 = 31.19 microg). Central injection of such muscarinic receptor antagonists did not affect tacrine-induced bradycardia. Our results show that acetylcholinesterase inhibitors induce significant cardiovascular effects with a pressor response mediated mainly by the stimulation of central muscarinic M2 receptors inducing a secondary increase in sympathetic outflow and vasopressin release. Conversely, acetylcholinesterase inhibitor-induced bradycardia appears to be mediated by peripheral muscarinic mechanisms. PMID- 9851544 TI - Functional characterisation of the pharmacological profile of the putative alpha1B-adrenoceptor antagonist, (+)-cyclazosin. AB - We studied the functional pharmacological profile of (+)-cyclazosin, which has been characterised as a selective, high-affinity (pKi = 9.68) alpha1B adrenoceptor ligand in binding experiments with rat liver membranes. The pKa/pA2 values for antagonism of contractions mediated via alpha1A/L-adrenoceptors of rat small mesenteric artery, alpha1B-adrenoceptors of rat aorta and alpha1B adrenoceptors of rat spleen were 7.78 +/- 0.04, 6.86 +/- 0.07 and 7.96 +/- 0.08, respectively. Furthermore, in mouse spleen, which is also regarded as an alpha1B adrenoceptor preparation, (+)-cyclazosin displayed low potency and did not act as a competitive antagonist. Thus, in contrast with results obtained in radioligand binding experiments, (+)-cyclazosin does not behave as a selective alpha1B adrenoceptor antagonist in functional tissues. Whether this discrepancy has consequences for the classification of alpha1-adrenoceptors requires further investigation. PMID- 9851546 TI - Modulation by lipid mediators of immune complex-induced lung inflammation in mice. AB - The present study characterized a murine model of immune complex-induced pneumonitis and investigated the role of platelet-activating factor (PAF) and eicosanoids as mediators of lung neutrophil infiltration and hemorrhagic lesions. Rabbit antibodies to bovine serum albumin were injected into the airways and bovine serum albumin was injected intravenously into C3H/HePas and BALB/c mice. After 24 h, a significant increase in neutrophil infiltration and hemoglobin concentration in the bronchoalveolar lavage fluid and lung parenchyma was observed in both strains despite the C3H/HePas strain being 10 times more sensitive to PAF. Neutrophil influx and vascular lesions were not affected by pre treatment of the mice with the PAF receptor antagonist, WEB 2170 (5-(2 chlorphenyl)carbonyl)-3,4-dihydro- 10-methyl-3-((4-morpholinyl)-2H,7H cyclopenta(4,5)thieno(3,2-f)(1,2,4)-t riazolo-(4,3-a)(1,4)-diazepine). In contrast, neutrophil influx and vascular lesions were increased by the cyclo oxygenase inhibitor, indomethacin, and reduced by the inhibitor of leukotriene synthesis, MK 886 (3-[1-(4-chlorobenzyl-3-t-butyl-thio-t-isopropyl-indol-2y-1]-2 2-+ ++dimethylpropanoic acid) and by the leukotriene B4 receptor antagonist, RO 0254094 (2-[(5-carboxypentyl)-6-[6-[3,4-dihidro-4-oxo-8-propyl-2H-1-benzop yran-7 yl)hexyl] benzenepropanoic acid). Increased levels of leukotriene B4, leukotriene C4/D4, thromboxane B2 were found in bronchoalveolar lavage fluid 4 h after induction of the reaction. There is also a tendency to increased prostaglandins E2 levels. Neutrophil infiltration and vascular lesions in immune complex-induced pneumonitis in mice are mediated by leukotriene B4. PMID- 9851545 TI - A possible role of S-nitrosothiols at the nitrergic relaxations in the mouse corpus cavernosum. AB - Relaxations induced by electrical field stimulation and acetylcholine were compared with those induced by acidified sodium nitrite, sodium nitroprusside, S nitrosoglutathione and S-nitroso-N-acetyl-D,L-penicillamine in the mouse corpus cavernosum precontracted with phenylephrine. NG-nitro-L-arginine inhibited electrical field stimulation- or acetylcholine-induced relaxation, but was ineffective on relaxations caused by the other stimuli. Hydroquinone and pyrogallol had no inhibitory action on the relaxations caused by any stimulus except acidified sodium nitrite. Incubation of the tissue with diethyldithiocarbamic acid significantly inhibited the relaxations induced by all stimuli except papaverine. In the tissues pre-treated with diethyldithiocarbamic acid, superoxide dismutase, hydroquinone and pyrogallol failed to yield restore or further inhibit the relaxations in response to electrical field stimulation or acetylcholine. LY 83583 (6-anilino-5,8-quinolinedione) and hydroxocobalamin clearly inhibited the relaxant responses to electrical field stimulation, acetylcholine, S-nitrosoglutathione and acidified sodium nitrite whereas there was significant enhancement of the relaxation produced by S-nitroso-N-acetyl-D,L penicillamine. These findings suggest that the relaxant factor released from non adrenergic non-cholinergic nerves or endothelial cells in mouse cavernosal tissue may be a superoxide anion-resistant nitric oxide-containing molecule and that S nitrosoglutathione rather than S-nitroso-N-acetyl-D,L-penicillamine could be a suitable candidate for this. PMID- 9851547 TI - Sulfatide reduces leucocyte accumulation and reverts vascular failure in splanchnic artery occlusion shock. AB - Selectin-mediated leucocyte accumulation is implicated in the pathogenesis of splanchnic artery occlusion. Sulfatide is recognized by P-selectin and blocks this adhesion molecule. We investigated the effects of sulfatide in rats subjected to splanchnic artery occlusion shock. Anaesthetized rats, subjected to total occlusion of the superior mesenteric artery and the coeliac trunk for 45 min developed severe shock resulting in death within 85-90 min after the release of occlusion. Sham operated animals were used as controls. Splanchnic artery occlusion shocked rats had marked hypotension, enhanced levels of tumor necrosis factor-alpha (TNF-alpha) in serum and macrophages, leucopenia and increased ileal leucocyte accumulation, studied by the means of myeloperoxidase activity. Furthermore, aortae from shocked rats showed marked hyporeactivity to phenylephrine (1 nM-10 microM), reduced responsiveness to acetylcholine (10 nM-10 microM) and an increased staining for P-selectin in the vasculature. In vivo administration of sulfatide (10 mg/kg, i.v., 5 min after occlusion of the splanchnic arteries) increased survival rate (90%, 4 h after splanchnic artery occlusion shock), enhanced mean arterial blood pressure, reduced serum TNF-alpha (37 +/- 11 U/ml vs. 398 +/- 18 U/ml), ameliorated leucopenia and reduced ileal myeloproxidase activity (1.2 +/- 0.4 U/g tissue vs. 8.2 +/- 0.8 U/g tissue). Aortae from splanchnic artery occlusion shocked rats treated with sulfatide exhibited a greater contractile response to phenylephrine and improved responsiveness to acetylcholine. Moreover sulfatide-treated rats showed a reduced staining for P-selectin in the aorta and in the superior mesenteric artery. Finally, passive immunization with specific monoclonal antibodies raised against P-selectin significantly protected from the lethality induced by splanchnic artery occlusion shock. Our results suggest that sulfatide protects against splanchnic artery occlusion shock. PMID- 9851548 TI - Cyclic GMP-dependent cytoprotection against ethanol-induced damage in rabbit isolated gastric parietal cells. AB - Prostaglandin E2 stimulates a nitric oxide/cyclic GMP (NO/cGMP) pathway which activates basolateral Cl- channels in rabbit gastric parietal cells. We examined whether the NO/cGMP pathway protects parietal cells from ethanol (EtOH)-induced cytotoxicity, using a parietal cell-rich suspension purified from rabbit gastric mucosa. Cytotoxicity was assayed by measuring the release of a fluorescent dye from the cells. N2,O2-dibutyryl guanosine 3',5'-cyclic monophosphate (DBcGMP) showed a concentration-dependent protective effect against EtOH-induced cytotoxicity. The half-maximal effect of DBcGMP was observed at 24 microM. DBcGMP in a concentration-dependent manner opened the basolateral Cl- channels of parietal cells, the EC50 value being 44 microM. The EtOH-induced cytotoxicity decreased as the Cl- concentration of medium decreased. A 30-s treatment with 5 nitro-2-(3-phenylpropylamino)-benzoate (NPPB), an inhibitor of the Cl- channel, had a cytotoxic effect which was not prevented by pre-incubation with DBcGMP. The cytotoxic effects of EtOH and NPPB were additive and the NPPB effects did not depend on the medium Cl- concentration. The present study showed that cGMP protects the gastric parietal cell from EtOH-induced cytotoxicity, and this cytoprotection is related to basolateral Cl- channel activity in the plasma membrane via an unknown mechanism(s). PMID- 9851549 TI - Modulation of the effects of extracellular ATP on [Ca2+]i in rat brain microvacular endothelial cells. AB - This study examined the intracellular regulation of signal transduction initiated by activation of the P2Y2 purinoceptor in a cultured rat brain microvascular endothelial cell line (RBE4). Intracellular free Ca2+ ([Ca2+]i) was monitored in single cells, using FURA-2 fluorimetry. As previously described [Nobles, M., Revest, P.A., Couraud, P.-O., Abbott, N.J., 1995. Characteristics of nucleotide receptors that cause elevation of cytoplasmic calcium in immortalized rat brain endothelial cells, RBE4, and in primary cultures. Br. J. Pharmacol., 115, 1245 1252], extracellular ATP (100 microM, 20 s) evoked a transient increase in intracellular free calcium concentration ([Ca2+]i). The amplitude of the Ca2+ transient evoked by ATP decreased with successive applications (desensitisation), as expected for a P2 purinoceptor. The modulation of the Ca2+ signal downstream to the activation of the ATP receptor was investigated, using agents selected for their ability to interfere with the intracellular pathways activated by ATP. The amplitude of the Ca2+ transient observed on the second application of ATP was compared in the presence and absence of these agents. The Ca2+ transient triggered by ATP was decreased by the inhibitor of nitric oxide synthesis, N omega-nitro-L-arginine methyl ester (L-NOARG). The inhibition induced by 100 microM L-NOARG was reversed by coapplication of the permeant cGMP analogue 8 brcGMP (100 microM). 8-BrcGMP caused a transient increase in [Ca2+]i when applied alone, and a dose-dependent inhibition of the increase in [Ca2+]i elicited by ATP. Indomethacin, an inhibitor of prostaglandin synthesis, inhibited the response to ATP. The inhibition caused by 10 microM indomethacin was reversed by coapplication of the permeant analogue of cAMP, 8-brcAMP (100 microM). 8-BrcAMP caused a transient rise in [Ca2+]i when applied alone, and a dose-dependent inhibition of the Ca2+ response evoked by ATP. The non-permeant cyclic nucleotides cAMP and cGMP did not affect the desensitising response to ATP, nor did they reverse the inhibitory actions of L-NOARG or indomethacin. It is concluded that cyclic nucleotides, nitric oxide, and prostaglandin synthesis pathways are able to interact with the Ca2+ second messenger pathway in rat brain endothelial cells activated by extracellular ATP. PMID- 9851550 TI - Dopamine D1 and D2 receptor selectivities of phenyl-benzazepines in rhesus monkey striata. AB - Several phenyl-benzazepine compounds, putatively selective dopamine D1 receptor agonists, have been used to study the effects of dopamine D1 receptor stimulation in rodents and nonhuman primates. However, the dopamine receptor selectivities of these compounds have not been established in nonhuman primates. Accordingly, the relative selectivities of six phenyl-benzazepines for dopamine D1-like and D2 like receptors were assessed in rhesus monkey and, for comparison, rat striata. The compounds tested had higher affinity for D1 than D2 receptors in both species; however, their selectivity varied by up to three orders of magnitude. GTP (100 microM) reduced agonist binding at the high-affinity state of the dopamine D1 receptor, but the magnitude of the effect of GTP did not reliably predict a compound's efficacy. Furthermore, a history of cocaine self administration did not appear to influence dopamine receptor binding characteristics in the rhesus monkeys in this study. The present results will aid the comparison of dopamine receptor binding characteristics and behavioral effects of D1 dopamine receptor agonists. PMID- 9851551 TI - Fluvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase, scavenges free radicals and inhibits lipid peroxidation in rat liver microsomes. AB - We investigated the effect of fluvastatin sodium (fluvastatin) and pravastatin, 3 hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, on the formation of thiobarbituric acid reactive substances both in vivo and in vitro in rat liver microsomes and on active oxygen species. Oral administration of fluvastatin at low doses (3.13 and 6.25 mg/kg) inhibited the formation of thiobarbituric acid reactive substances in rat liver microsomes, but high doses (12.5 and 25 mg/kg) did not change the formation of thiobarbituric acid reactive substances. Fluvastatin at any dose used had no effect on the content of cytochrome P-450 and the activity of NADPH-cytochrome P-450 reductase. In in vitro experiments, concentrations of fluvastatin ranging from 1 x 10(-6) - 1 x 10(-4) M markedly inhibited NADPH-dependent lipid peroxidation in liver microsomes, but pravastatin weakly inhibited lipid peroxidation. The order of magnitude of inhibition of each drug on in vitro lipid peroxidation was butylated hydroxytoluene > probucol > or = fluvastatin > pravastatin. Moreover, fluvastatin chemically scavenged active oxygen species such as hydroxyl radicals and superoxide anion generated by the Fenton reaction and by the xanthine-xanthine oxidase system, respectively, but pravastatin showed no scavenging of superoxide anion. These results indicate that the suppression of in vivo and in vitro lipid peroxidation in liver microsomes may be, at least in part, due to the scavenging by fluvastatin of free radicals. PMID- 9851552 TI - Furosemide and digoxin inhibit thiamine uptake in cardiac cells. AB - Heart cells in culture were used to clarify whether furosemide or digoxin cause thiamine deficiency and if so, by what mechanism. The intracellular level of thiamine pyrophosphate gradually decreased, with a half-life of 16-19 days, after treatment of cardiac cells with furosemide or digoxin. When thiamine was excluded from the growth medium, thiamine pyrophosphate levels gradually decreased, with a half-life of 5-6 days. No additive effect was observed in the presence of the above drugs when thiamine was excluded from the medium. Thiamine uptake by cardiac cells grown in a thiamine-free medium for 7 days decreased significantly in the presence of furosemide or digoxin. The effect of furosemide or digoxin on thiamine uptake was found to be dose dependent. Co-administration of furosemide and digoxin to the cardiac cell cultures resulted in an additive effect on thiamine uptake. Our results demonstrate that furosemide and digoxin inhibit thiamine uptake by cardiac cells in culture and may therefore cause thiamine deficiency in patients undergoing chronic treatment with these drugs. PMID- 9851553 TI - Sigma-binding site ligands inhibit K+ currents in rat locus coeruleus neurons in vitro. AB - Biological actions of novel sigma1- and sigma2-selective binding site ligands (trishomocubanes: 4-azahexacyclo [5.4.1.0.(2,6).0(3,10).0(5,).0(8,11)]dodecanes), and the reference ligands, 1,3-di(2-tolyl)-guanidine (DTG), haloperidol, (+) pentazocine and dextromethorphan, were studied in rat locus coeruleus neurons using intracellular and whole-cell patch clamp recordings. High concentrations of trishomocubanes produced small inward currents and affected some parameters of action potential waveforms suggesting modest potency to inhibit ionic conductances underlying action potentials. Sigma-ligands produced large inward currents in the presence of mu-opioid, alpha2-adrenoceptor and ORL1 receptor agonists. These reversed polarity near the K+ equilibrium potential, suggesting that sigma-ligands act as ligand activated K+-channel blockers or interfere with the coupling between these receptors and K+-channels. However, no correlation was found between binding affinities at sigma1- or sigma2-binding sites and potency to inhibit K+-currents, suggesting that these effects on K+-channels are not directly related to occupancy of sigma binding sites. PMID- 9851554 TI - Recent advances in imidazoline receptor research: ligands--localization and isolation--signaling--functional and clinical studies. AB - In this article we outline the highlights of this special issue of the journal containing a series of articles covering many aspects of current interest in the field of imidazoline receptor research. This volume is the result of an international symposium held in September 1997 in Melbourne as an official satellite of the inaugural meeting of the International Society of Autonomic Neurosciences held in Cairns, Australia. A wide range of topics relating to imidazoline receptors were canvassed, including endogenous and synthetic ligands, identification and localisation of binding sites, putative transduction mechanisms and experimental and clinical functional studies. PMID- 9851555 TI - Agmatine: an endogenous ligand at imidazoline receptors may be a novel neurotransmitter in brain. AB - Agmatine, which in other life forms serves as a metabolic intermediate for polyamine biosynthesis, appears to have properties in mammals consistent with its actions as a neurotransmitter/neuromodulator. Thus, agmatine is synthesized unequally in brain by arginine decarboxylase (ADC); is stored in neurons and axon terminals with a heterogeneous distribution; is released from synaptosomes by depolarization; is enzymatically converted by agmatinase to putrescine; interacts not only with alpha2-adrenergic and I-receptors in the CNS, but also may selectively block NMDA receptor channels; and, when administered centrally, has several potent biological actions. Clarification of its role in normal brain function, however, has not yet been fully established, in part because of the absence of agents that selectively affect its biosynthesis or degradation. PMID- 9851556 TI - Isolation and partial structure determination of a clonidine-displacing substance from bovine lung and brain. AB - A large scale extraction and isolation method was developed for the purification of clonidine-displacing substance (CDS) activity from bovine lung or brain. This optimised method used direct freeze drying of tissue, hexane removal of lipids, and methanol extraction of CDS activity. Using a bioassay directed isolation strategy a new CDS compound was purified from an extract of bovine lung. The isolation strategy involved subsequent steps of flash C-18 chromatography, ion exchange, size exclusion, and C-18 HPLC. An HPLC detection method was developed and applied to show that the new CDS is present in both lung and brain tissue. Spectroscopic data for this new CDS indicates that it is related to guanosine, but is not noradrenaline, adrenaline, histamine, agmatine, guanosine, GMP, GDP or GTP. PMID- 9851557 TI - Does a second generation of centrally acting antihypertensive drugs really exist? AB - The site of the hypotensive action of imidazoline compounds, such as clonidine, was first identified within the rostroventrolateral part of the brainstem: the nucleus reticularis lateralis. After that, it was shown that imidazolines and related substances reduced blood pressure when applied in this area whereas catecholamines were not capable of producing such an effect. These data led us to suggest the existence of receptors specific for imidazoline-like compounds different from the alpha2-adrenoceptors. Soon after, the existence of imidazoline binding sites was reported in the brain and in a variety of peripheral tissues including the human kidney. As expected, these specific binding sites do not bind the catecholamines. The imidazoline binding sites are already subclassified in two groups: the I1-subtype sensitive to clonidine and idazoxan, and the I2 subtype, sensitive to idazoxan and nearly insensitive to clonidine. Functional studies confirmed that the hypotensive effects of clonidine-like drugs involved imidazoline receptors while their most frequent side effects only involved alpha2 adrenoceptors. However, recent functional evidence suggests that a cross talk between imidazoline receptors and alpha2-adrenoceptors is necessary to trigger a hypotensive effect within the ventral brainstem. Rilmenidine and Moxonidine are the leader compounds of a new class of antihypertensive drugs selective for imidazoline receptors. At hypotensive doses, these drugs are devoid of significant sedative effect. Rilmenidine evoked hypotension when injected within the nucleus reticularis lateralis region; it competed for [3H]-clonidine bound to specific imidazoline binding sites in human medullary membrane preparations but proved more selective for cerebral imidazoline receptors than clonidine. It is suggested that this selectivity might explain the low incidence of their side effects. Additional potentially beneficial actions on cardiac arrhythmias or congestive heart failure enlarge the therapeutic interest of imidazoline-related drugs. Recent binding and functional data throw a new light on the optimal pharmacological profile of this second generation of centrally acting antihypertensive drugs. PMID- 9851558 TI - Characterization of a partial cDNA clone detected by imidazoline receptor selective antisera. AB - A cDNA clone has been isolated from a human hippocampal cDNA expression library by relying on the selectivity of two antisera that are specific for imidazoline binding proteins. A 1789 bp cDNA clone was sequenced and shown to contain a single open-reading frame that predicts a 66 kDa polypeptide, but it is truncated based on its lack of a stop codon and poly-A+ tail. Two regions of homology exist for the predicted amino acid sequence in common with chromogranin-A and B proteins, a zinc finger protein, and the ryanodine receptor. Northern blot analyses of poly-A+ mRNA from 36 human tissues indicated two differentially expressed transcripts of 6.0 and 9.5 kb. The 6.0 kb mRNA form was enriched in brain and endocrine tissues as compared to other tissues, but not in strict concordance with I1-imidazoline binding sites. The highest overall amounts of the combined transcripts were found in pituitary. In situ hybridization histochemistry revealed an enrichment of the message in neuronal cell bodies of the rat hippocampus and cerebellar cortex. This clone has some of the properties expected of an imidazoline receptor. PMID- 9851559 TI - Characterization of [3H]idazoxan binding proteins in solubilized membranes from rabbit and human liver. AB - Several studies have shown that I2 imidazoline binding sites are localized on monoamine oxidases. Recent results obtained after solubilization of rat brain membranes and analysis by size-exclusion chromatography suggested the existence of additional I2 imidazoline binding sites located on proteins distinct from monoamine oxidases. In order to define whether such binding sites are expressed in human and rabbit liver, we solubilized I2 imidazoline binding sites and monoamine oxidases and compared their elution profile by size-exclusion chromatography. I2 binding sites were labeled using [3H]idazoxan. Monoamine oxidases were identified by the measure of [14C]tyramine oxidation and Western blot analysis using an anti-MAO-A/MAO-B polyclonal antiserum. After solubilization of rabbit or human liver using 1% digitonin, 90% of [3H]idazoxan binding eluted in a major peak corresponding to a Mr of approximately 175000 Da. A minor peak, (Mr approximately equal to 100000 Da) representing 10% of the recovered [3H]idazoxan binding, was also observed. [14C]tyramine oxidation as well as immunoreactive bands corresponding to MAOs were exclusively detected in fractions containing [3H]idazoxan binding. These results show that solubilized I2 imidazoline binding sites distinct from monoamine oxidases are not detectable in rabbit and human liver. PMID- 9851560 TI - [3H]Rilmenidine-labelled imidazoline-receptor binding sites co-localize with [3H]2-(benzofuranyl)-2-imidazoline-labelled imidazoline-receptor binding sites and monoamine oxidase-B in rabbit, but not rat, kidney. AB - The distribution and relative densities of imidazoline-receptor binding sites (I RBS) and monoamine oxidase (MAO)-A and -B enzyme(s) in rat and rabbit kidney were compared autoradiographically using fixed nanomolar concentrations of [3H]rilmenidine and [3H]2-(benzofuranyl)-2-imidazoline ([3H]2-BFI) to label I RBS, and [3H]RO41-1049 and [3H]RO19-6327 to label MAO-A and -B isoenzymes, respectively. In rat kidney, high densities of I-RBS labelled by [3H]rilmenidine were observed in the cortex and outer stripe (120-280 fmol/mg tissue), in contrast to low I-RBS densities labelled by [3H]2-BFI (<4 fmol/mg). A relatively high density of [3H]RO41-1049 binding to MAO-A enzyme was present in all regions of the rat kidney (160-210 fmol/mg) compared with a low density of [3H]RO19-6327 binding to MAO-B (< 25 fmol/mg). Comparison of MAO-A and -B distributions with that of [3H]rilmenidine-labelled I-RBS strongly suggests a lack of association in rat kidney. Similarly, the extremely low densities of [3H]2-BFI-labelled I2-RBS in rat kidney contrasts with the density of MAO-A, but is consistent with the low density of MAO-B. Rabbit kidney cortex and outer stripe contained high relative densities of [3H]rilmenidine-labelled I-RBS (200-215 fmol/mg) and [3H]2-BFI labelled I2-RBS (45-60 fmol/mg) with lower densities in the inner stripe and inner medulla (< or = 100 and 30 fmol/mg respectively). A high density of MAO-A binding was observed in the inner stripe (515 fmol/mg) with lower levels in the cortex and outer stripe (100-240 fmol/mg), while high densities of MAO-B binding were observed in the cortex and outer stripe (290-450 fmol/mg) with lower levels in the inner stripe (65 fmol/mg). The correlation between the localization of [3H]rilmenidine-labelled I-RBS and [3H]RO19-6327-labelled MAO-B in rabbit kidney (r = 0.87, P = 0.057) suggest that [3H]rilmenidine may label a binding site co existent with MAO-B, but not MAO-A (n.s.), in this tissue, but rilmenidine did not inhibit [3H]RO41-1049 or [3H]RO19-6327 binding. The distribution of [3H]2-BFI labelled I2-RBS overlapped the combined distributions of both MAO-A and -B isoenzymes, suggesting that [3H]2-BFI may label sites on both enzymes in the rabbit, but [3H]2-BFI binding only correlated with [3H]RO19-6327 (r = 0.84, P = 0.07), not [3H]RO41-1049 binding (n.s.). Moreover, 2-BFI only inhibited [3H]RO19 6327, not [3H]RO41-1049 binding. These data are consistent with reports that I2 RBS are located on MAO-B and allosterically influence the catalytic site. The relationship of [3H]rilmenidine- and [3H]2-BFI-labelled I-RBS and the identity of non-MAO-associated [3H]rilmenidine-labelled I-RBS requires further investigation. PMID- 9851562 TI - Investigation of I1-imidazoline receptors using microphysiometry and molecular modelling. AB - The molecular identity and structure of imidazoline receptors is still poorly understood. For example the I1-imidazoline binding site (I1-site) is localised to the plasma membrane, but it is not clear if this represents a conventional receptor. The I1-site reportedly has both high and low affinity binding states. Again it is not clear if these sites represent different states of the same receptor, or distinct molecular entities. The signal transduction mechanisms of I1-imidazoline receptors are beginning to be unravelled. There is clear evidence that ligands with high affinity for I1-sites stimulate phosphatidylcholine selective phospholipase C in the rat adrenal medullary tumour cell line PC-12, but this may not be the case in all cell types. We investigated the possible role of this novel pathway in bovine adrenal medullary cells. Radioligand binding studies with [3H]clonidine confirmed the presence of I1-sites in membranes from these cells. Using microphysiometry, a recently developed technique for determining cellular activation, the extracellular acidification rates of cultured bovine adrenal medullary cells were unaffected by a number of imidazolines considered to be agonists at the I1-site. This suggests that there is no I1-site mediated stimulation of phosphatidylcholine specific phospholipase C in these cells. However, nicotine-stimulated increases in extracellular acidification were blocked by 100 microM clonidine. Ion channels have been suggested as another possible I1-imidazoline 'receptor' family, and may represent the low affinity I1-site detected in binding studies. I1-Site ligands can be shown to bind to, or block, several members of the ligand-gated ion channel superfamily, including the 5HT3, K+ATP, NMDA and nicotinic acetylcholine receptors. The I1-site ligands appear to be binding to, and acting at, the previously described phencyclidine binding site in these channels. Furthermore, molecular modelling suggests that I1-site selective ligands share a common three dimensional structure with phencyclidine, and that I2-site selective ligands do not have this structure. This suggests that a phencyclidine-binding site motif may represent a novel site of action for I1-site ligands, and a search for receptors based on this motif may reveal novel imidazoline 'receptors'. PMID- 9851561 TI - Pharmacology and subcellular distribution of [3H]rilmenidine binding sites in rat brain. AB - We have previously reported that in rat brain membranes, [3H]rilmenidine, in addition to labelling alpha2-adrenoceptors and the I2B-subtype of imidazoline receptor binding site (I2B-RBS), may label an additional I-RBS population, distinct from previously classified I1-RBS and I2-RBS. In this study, using crude or fractionated rat brain membranes we examined the possible association of [3H]rilmenidine-labelled I-RBS with the A- and B-isoforms of monoamine oxidase (MAO) by studying the inhibition of [3H]rilmenidine binding by a number of MAO inhibitors; and comparing the maximal binding density (Bmax) and subcellular distribution of [3H]rilmenidine binding sites with that of MAO-A and MAO-B catalytic sites labelled by [3H]RO41-1049 and [3H]RO19-6327 and 12-RBS labelled by [3H]2-BFI. Inhibition of [3H]rilmenidine binding by all MAO inhibitors tested produced very shallow curves (slope 0.29-0.56). Clorgyline and moclobemide (selective MAO-A inhibitors) displayed moderate affinities (60-140 nM), while pargyline (non-selective MAO-inhibitor), RO41-1049 (selective MAO-A inhibitor) and RO19-6327 (selective MAO-B inhibitor) exhibited very low affinities (> 2 microM) for 50-75% of [3H]rilmenidine-labelled I-RBS in crude brain membranes and even lower affinity for the remaining binding. Under identical buffer conditions, the Bmax of [3H]rilmenidine-labelled I-RBS (1.45+/-0.14 pmol/mg protein) was considerably lower than those of MAO-A (13.10+/-0.15 pmol/mg) and MAO-B (10.35+/ 0.50 pmol/mg) sites. These results suggest that [3H]rilmenidine does not interact directly with the active catalytic site of either MAO enzyme and could at best only associate with a subpopulation of MAO molecules. Binding studies on five fractions of rat cortex homogenates-nuclear (N), heavy (M) and light (L) mitochondrial, microsomal non-mitochondrial (P), and soluble cytosolic (S) fractions-revealed that 45% of total [3H]rilmenidine binding was present in the P fraction cf. 20 and 23% in the M and L fractions, in contrast to [3H]RO19-6327 and [3H]2-BFI which bound 11-13% in the P fraction and 36-38% and 35-44% in the M and L fractions, respectively. Binding of all ligands in the N fraction was 6-15% of total. These studies reveal that [3H]rilmenidine-labelled I-RBS, unlike the I2 RBS, are not predominantly associated with mitochondrial fractions containing the MAO enzymes (and cytochrome oxidase activity), but appear to be distributed in both the mitochondrial and plasma membrane fractions in rat cerebral cortex. PMID- 9851563 TI - Arachidonic acid release from PC12 pheochromocytoma cells is regulated by I1 imidazoline receptors. AB - Rat PC 12 pheochromocytoma cells lack alpha2-adrenergic receptors but express plasma membrane I1-imidazoline receptors. In response to the I1-agonist moxonidine, diglycerides are generated via phosphatidylcholine-selective phospholipase C, and prostaglandin E2 is released. This report characterizes I receptor-mediated release of arachidonic acid, the precursor to the prostaglandins. PC12 cells were incubated with [3H]arachidonic acid for 24 h and superfused with 0.01% bovine serum albumin in Krebs' physiological buffer at 1 ml/min. Calcium ionophore increased arachidonic acid release only marginally, implying that in PC12 cells arachidonic acid release is not driven by calcium. The I1-agonist moxonidine at concentrations between 10 nM and 1.0 microM rapidly elicited up to two-fold increases in [3H]arachidonic acid release. Guanabenz, a potent alpha2-agonist and I2-ligand, had no effect. The selective I1-antagonist efaroxan blocked the action of moxonidine. The phospholipase A2 inhibitor aristolochic acid had no effect, suggesting that arachidonic acid release may be through an indirect pathway, possibly involving diglycerides. Thus, I1 imidazoline receptors in PC12 cells are coupled to arachidonic acid release through an as yet unknown pathway. PMID- 9851564 TI - Imidazoline receptor mediated natriuresis: central and/or peripheral effect? AB - The ability of imidazoline agonists, such as moxonidine and rilmenidine, to lower blood pressure has been attributed to a central effect resulting in a decrease in peripheral sympathetic nerve activity. A similar decrease in sympathetic nerve activity to the kidney has been proposed to explain the increase in sodium excretion. The observed increase in sodium excretion following an intrarenal infusion of moxonidine or rilmenidine suggested the existence of a direct renal action. We therefore tested the hypothesis that direct renal infusions were acting at a central rather than a peripheral site. Thus, interventions which would decrease the natriuretic effects of central administered moxonidine would also block the effects of intrarenal administered moxonidine. Studies were performed in anesthetized Sprague-Dawley rats (280-320 g) which had undergone unilateral nephrectomy 7 to 10 days prior to the experiment. The interventions utilized resulted in minimal effects on blood pressure and creatinine clearance. Intracerebroventricular (icv) or intrarenal (ir) administration of moxonidine produced a significant increase in urine flow rate and sodium excretion. Intravenous (iv) prazosin was used to block the ability of the sympathetic nerves to alter sodium excretion secondary to alpha1-adrenoceptor stimulation. Prazosin prevented the natriuresis following icv moxonidine but only partially antagonized the effects of ir moxonidine. To determine if central imidazoline receptors mediated the effects of moxonidine, animals were pretreated with icv idazoxan. Following icv idazoxan, the effects of icv moxonidine were blocked, whereas the response to intrarenal moxonidine was only partially blocked. Peripheral (iv) administration of idazoxan blocked the actions of intrarenal moxonidine but left the response to icv moxonidine intact. Finally, chemical sympathectomy with reserpine did not alter the response to intrarenal moxonidine suggesting that this effect was independent of the sympathetic nervous system. In conclusion, these studies indicate the ability of central and peripheral moxonidine to increase urine flow rate through sodium excretion at two unique sites of action, one central and the other one peripheral, most conceivably within the kidney. PMID- 9851565 TI - Relationship between imidazoline and alpha2-adrenoceptors involved in the sympatho-inhibitory actions of centrally acting antihypertensive agents. AB - Since the first suggestion of the existence of imidazoline receptors, there has been a continuing and yet unresolved debate as to their contribution to the antihypertensive actions of clonidine-like agents. In this review we bring together a number of studies from our laboratory which have examined the importance and interdependence of imidazoline receptors and alpha2-adrenoceptors in the mechanism of action of centrally acting antihypertensive drugs. Using conscious rabbits and a range of imidazoline and specific alpha2-adrenoceptor antagonists we have consistently found that second generation agents rilmenidine and moxonidine preferentially act via imidazoline receptors but that alpha2 adrenoceptors are important for the hypotension produced by clonidine and alpha methyldopa. Despite this difference in receptor mechanism, the hypotension produced by all these drugs is dependent on central noradrenergic pathways. In other studies using anaesthetised rabbits and direct measures of sympathetic nerve activity we confirmed the generally held view that the major site of sympatho-inhibitory actions and sympathetic baroreflex effects of centrally acting antihypertensive agents is the rostral ventrolateral medulla (RVLM). We also found, using microinjection of specific antagonists, that alpha2 adrenoceptors in this nucleus appear to be activated as a consequence of imidazoline receptor activation. Thus, there appears to be a close relationship between imidazoline receptors and alpha2-adrenoceptors located in the RVLM in mediating the hypotension and inhibition of renal sympathetic nerve activity. Furthermore in recent studies using a noradrenergic neurotoxin microinjected into the RVLM we found that this treatment selectively blocked the actions of moxonidine but not clonidine, suggesting that I1-imidazoline receptors may be located on adrenergic terminals in situ. By contrast, clonidine acts predominantly via alpha2-adrenoceptors, perhaps located on cell bodies in the nucleus. We conclude that there is indeed a close nexus between 'presynaptic' imidazoline receptors on noradrenergic terminals and 'downstream' alpha2 adrenoceptors within the RVLM. Our hypothesis brings together opposing points of view that the mechanism for hypotension must involve either the imidazoline receptor or the alpha2-adrenoceptor. Clearly both are important. PMID- 9851566 TI - Chronopharmacological dependence of antihypertensive effects of the imidazoline like drugs in stroke-prone spontaneously hypertensive rats. AB - In the present study we investigated the chronopharmacological dependence of dose dependent hypotensive and cardiochronotropic effects of the imidazoline-like drugs (clonidine, rilmenidine and moxonidine) in stroke-prone spontaneously hypertensive rats (SHR-SP), using radio-telemetric system (Data Sciences, USA). The 24-h blood pressure, heart rate and locomotor activity profiles showed peak values during the rats' active phase during the night period. The degree of hypotensive and bradycardic effects of all drugs were most evident at this time and occurred in the absence of a change in locomotor activity. These studies show that clonidine, rilmenidine and moxonidine decrease blood pressure and heart rate in a time-dependent manner in SHR-SP. It was demonstrated that the degree and duration of hypotensive action of imidazoline-like drugs vary with the time of drug administration. PMID- 9851567 TI - Effect of rilmenidine on the cadiovascular responses to stress in the conscious rabbit. AB - Environmental stress can cause an increase in sympathetic nerve activity both in humans and animals. While centrally acting antihypertensive drugs such as rilmenidine are known to reduce sympathetic tone, it is not clear whether they also influence the cardiovascular responses to acute stress. In the present study we examined the effects of systemic treatment with rilmenidine on the sympathetic and haemodynamic responses to air jet or noise stress. Twelve conscious rabbits previously implanted with a renal nerve recording electrode were subjected to an 8 l/min stream of air directed at their face for 10 min or exposure to 10 min of white noise (approximately 85 dB). Both air jet and noise stress elicited increases in renal sympathetic nerve activity (RSNA) which were greatest in the first minute (+55+/-9% and +40+/-6%, respectively), but which quickly reached a stable level over the subsequent 9 min (+24+/-6% and +9+/-5%, respectively). This was accompanied by a small increase in heart rate (HR) and mean arterial pressure (MAP). Intravenous rilmenidine (273 microg/kg) reduced MAP from 85+/-3 mm Hg to 68+/-2 mm Hg and HR from 203+/-10 b/min to 188+/-10 b/min and lowered basal RSNA by 54%. Rilmenidine reduced the increase in RSNA seen during the first minute of air jet stress by 35% and reduced the average increase over the next 9 min by 68%. However, rilmenidine had little effect on either the initial or stable RSNA responses to noise stress. Saline treatment did not alter the RSNA responses to either air jet or noise stress. The results show that centrally-acting antihypertensive agents not only lower basal RSNA, but can differentially influence environmentally induced sympathetic responses. In addition, the differential effect of rilmenidine on noise and air jet stress suggests that they may involve quite different central processing. PMID- 9851568 TI - Interaction of the dopamine D2 receptor agonist quinpirole with sympathetic vasomotor tone and the central action of rilmenidine in conscious rabbits. AB - Previous studies in conscious rats have shown that systemic administration of the dopamine D2 receptor agonist quinpirole causes a centrally-mediated increase in blood pressure which is associated with increased plasma levels of noradrenaline and adrenaline. In addition, treatment with quinpirole caused a marked inhibition of the antihypertensive effect of centrally-acting sympatho-inhibitory drugs such as clonidine, rilmenidine and alpha-methyldopa, suggesting an interaction at the level of sympathetic vasomotor tone. The main aim of the present study was investigate in conscious rabbits the effect of quinpirole on renal sympathetic nerve activity. In addition, we studied the effect of pretreatment with quinpirole on responses to additional quinpirole injections or rilmenidine treatment. Quinpirole treatment caused a prolonged dose-dependent increase in blood pressure and heart rate. Additional injection of quinpirole, 30 min after the first treatment, caused a significantly smaller pressor response (7+/-2 vs. 17+/-2 mm Hg). Injection of rilmenidine caused a larger decrease in blood pressure in rabbits which had been pretreated with quinpirole than in controls ( 28+/-3 vs. -14+/-3 mm Hg). Total renal sympathetic nerve activity was markedly increased by quinpirole treatment (3.5-fold), an effect which could be attributed to both increased amplitude and increased frequency of the renal nerve signal. After a second injection of quinpirole, 30 min after the first treatment, only total renal sympathetic nerve activity and amplitude were increased and the effects were reduced. These results show marked actions of quinpirole on renal sympathetic nerve activity in conscious rabbits. However, the previously described apparent desensitisation to the antihypertensive effect of rilmenidine could not be observed in rabbits, suggesting marked species differences in the mechanism and site of action of rilmenidine. PMID- 9851569 TI - Comparison of the baroreceptor-heart rate reflex effects of moxonidine, rilmenidine and clonidine in conscious rabbits. AB - In 10 conscious rabbits, the baroreceptor-heart rate (HR) reflex effects of centrally acting antihypertensive agents with high affinity for imidazoline receptors (IRs), moxonidine and rilmenidine, were compared with clonidine which acts predominantly via central alpha2-adrenoceptors. Dose regimens were chosen to give similar hypotension (-17+/-1 mm Hg) and bradycardia (-27+/-2 b/min) for all three agents given into the fourth ventricle. Baroreceptor-HR reflex curves were assessed by i.v. drug induced changes in blood pressure. With all treatments, the baroreflex curves with both vagal and sympathetic effectors intact were shifted to the left, corresponding to the hypotension, and the bradycardia plateau was reduced. Rilmenidine and moxonidine also reduced the upper plateau such that the curves were shifted parallel down the HR scale with no change in the HR range. By contrast, clonidine only decreased the lower plateau, and thus increased HR range (+19+/-6%). Moxonidine, but not rilmenidine, reduced the baroreflex gain by reducing the curvature. Clonidine also decreased curvature but this did not result in a reduction in gain as it was offset by the increase in HR range. The gain and range of the cardiac sympathetic component, as assessed after vagal blockade, was reduced by rilmenidine by 53 and 40% respectively, but was not affected by the other agents. The calculated vagal component of the curves showed that all agents produced a greater vagal bradycardia in response to a rise in pressure and that both rilmenidine and clonidine increased vagal HR range. The present study results show that many of the baroreflex effects of clonidine, such as facilitating cardiac vagal responses, are shared by the second generation agent rilmenidine, suggesting that they are primarily due to alpha2-adrenoceptor activation. In addition, the inhibition of the sympathetic component of the baroreflex, observed with rilmenidine, and not clonidine suggests that this effect may involve IRs. By contrast moxonidine, the most specific agent for I1 receptors, produces mainly a baroreflex independent inhibition of cardiac sympathetic activity with little effect on vagal activity. PMID- 9851570 TI - Influence of sino-aortic barodenervation on the cardiovascular effects of imidazoline-like drugs. AB - Earlier findings have shown that hypotensive effects of centrally acting drugs, such as clonidine, are enhanced in animals after denervation of arterial baroreceptors. The purpose of this study was to investigate the dynamic of changes in arterial pressure, heart rate and hypotensive effects of clonidine, rilmenidine and moxonidine in Wistar rats after sino-aortic denervation (SAD) using radio-telemetry. SAD was followed by significant elevation of arterial pressure lability (the standard deviation of the mean arterial pressure), while the baseline mean arterial pressure (MAP) and heart rate in barodenervated rats (12 days after SAD) was similar to intact rats. The hypotension produced by clonidine, rilmenidine and moxonidine was much greater in SAD rats than in intact rats. The study suggests that baroreflex mechanisms are not only important for maintaining levels of blood pressure in the very short term, but also for buffering the effects of centrally acting antihypertensive drugs. PMID- 9851571 TI - Increased sympathetic nervous system activity and its therapeutic reduction in arterial hypertension, portal hypertension and heart failure. AB - Although the underlying mechanisms no doubt differ, activation of the sympathetic nervous system is an important pathophysiological feature in primary arterial hypertension, in portal hypertension accompanying hepatic cirrhosis, and in heart failure, and is a logical therapeutic target for centrally acting sympathetic nervous system suppressant drugs. Portal hypertension: The sympathetic outflows to skeletal muscle vasculature, the heart, the kidneys and to the hepatomesenteric circulation are stimulated in patients with alcoholic cirrhosis of the liver, perhaps as a reflex response to the vasodilatation and vascular shunting present. Acute dosing with clonidine produces dose dependent reduction in noradrenaline spillover from visceral organs and reduction in hepatic vein wedge pressure, with preservation of hepatic blood flow and negligible fall in arterial pressure. These findings indicate the clinical potential of drugs such as clonidine, moxonidine and rilmenidine for chronically lowering portal venous pressure in cirrhosis. Arterial hypertension: Activation of the sympathetic outflow to the heart, kidneys and skeletal muscle vasculature is commonly present in younger (< 45 years) patients with essential hypertension. The sympathetic stimulation appears to have adverse consequences in hypertensive patients beyond blood pressure elevation. Neural vasoconstriction in skeletal muscle has metabolic effects by impairing glucose delivery, which is a basis for insulin resistance and hyperinsulinemia. Within the heart a trophic effect of sympathetic activation on cardiac growth, contributing to the development of left ventricular hypertrophy, and an arrhythmogenic effect are also likely. Cardiac failure: The cardiac sympathetic nerves are preferentially stimulated in severe heart failure, with norepinephrine release from the failing heart at rest being increased as much as 50-fold, similar to the level seen in healthy people during near maximum exercise. This preferential activation of the cardiac sympathetic outflow contributes to arrhythmogenesis and possibly to progression of the heart failure, and has been directly linked to mortality; a high rate of spillover of noradrenaline from the heart is a strong, independent predictor of poor prognosis in severe cardiac failure. The mechanisms underlying sympathetic nervous stimulation are not entirely clear. Increased intracardiac diastolic pressure seems to be one peripheral signal, and increased forebrain norepinephrine turnover an important central mechanism. Following the demonstration of the beneficial effect of the beta-adrenergic blocker, carvedilol, and with second generation centrally acting sympathetic suppressants now under clinical investigation, elucidation of the abnormalities in central nervous control of sympathetic outflow in heart failure has become clinically relevant. PMID- 9851572 TI - Social phobia: the nature of the disorder. AB - Social phobia is characterized by a fear of social situations, which are either avoided or endured with intense anxiety or distress. There is a high prevalence of social phobia in the community and the course of the condition tends to be chronic. In spite of the personal impact of social phobia, few sufferers seek professional help. This is due to a lack of awareness about the condition on the part of health professionals and the public. Social phobia demands increased awareness, so that sufferers receive the treatment they need and deserve, so giving them the opportunity to improve their quality of life. PMID- 9851573 TI - Subtypes of social phobia and comorbidity with depression and other anxiety disorders. AB - Epidemiological studies have identified two subtypes of social phobia: speaking only social phobia which is characterized by the fear of public speaking situations and complex social phobia which is characterized by the fear of multiple social situations. Speaking-only social phobia most closely corresponds to the DSM-IV's 'nongeneralized social phobia' while complex social phobia resembles 'generalized social phobia'. In contrast to the speaking-only social phobia, the complex form is usually more disabling, familial and longer-lasting. In addition, the complex form has a lower chance of spontaneous recovery and carries a higher risk of comorbidity and impairment. Overall, both types of social phobia tend to be underdiagnosed and under-treated. Effective treatments which can manage not only complex social phobia, but also its spectrum of comorbid conditions, are required. PMID- 9851574 TI - Implications of the severity of social phobia. AB - This paper presents a post hoc analysis of a recent large, 12-week, placebo controlled trial of paroxetine in the treatment of social phobia, which analyzes subgroups of patients classified as suffering from severe (Liebowitz Social Anxiety Scale [LSAS] total score > or = 82, n = 85) or moderate (LSAS total score 52-81, n = 78) social phobia. With respect to the reduction in LSAS total score, the paroxetine-placebo difference was greater in patients in the severe subgroup (20.0, P < 0.001) than those in the moderate subgroup (13.7, P < 0.02). Likewise, for the number of patients rated as 'very much' or 'much' improved according to their Clinical Global Impressions improvement scores, the paroxetine-placebo difference was greater in the severe subgroup (34.2%, P < 0.001) than in the moderate subgroup (29.1%, P < 0.02). In conclusion, paroxetine is effective compared with placebo in both moderate and severe social phobia, and the response is more clear cut in patients with more severe symptoms. PMID- 9851575 TI - Social phobia and alcoholism: a complex relationship. AB - The relationship between social phobia and alcoholism is complex. Alcohol problems typically develop secondary to social phobia, with patients reporting that they find alcohol helpful in coping with the symptoms of anxiety. However, excessive alcohol consumption may actually precipitate anxiety symptoms, and thus a vicious circle of anxiety and alcoholism is established. The clinician must consider the possibility of comorbidity in patients presenting with either alcoholism or social phobia. Only when both components of the condition are adequately assessed and treated can the cycle of social phobia and alcohol abuse be broken. PMID- 9851576 TI - The clinical relevance of treating social phobia. AB - Social phobia is an extreme fear of being scrutinized by others in social situations that often culminates in avoidance of those situations. Studies have demonstrated marked educational, occupational and social disabilities in patients with social phobia. However, many sufferers do not seek help and are not detected when seeking medical advice for other conditions. The advent of a range of effective treatments, in particular the selective serotonin reuptake inhibitors, has substantially improved the prognosis in these sufferers, both symptomatically and in terms of quality of life. PMID- 9851577 TI - The role of drug therapy in social phobia. AB - Social phobia is a potentially disabling condition requiring effective and well tolerated treatment. The monoamine oxidase inhibitors, benzodiazepines and, more recently, the selective serotonin reuptake inhibitors (SSRIs) have all shown promise as treatments for this disorder. Paroxetine, the most thoroughly studied SSRI, has been shown to be effective in treating the symptoms of generalized social phobia in a large-scale, placebo-controlled trial. Paroxetine and the other SSRIs show promise as a new and safe therapeutic choice for patients with social phobia. PMID- 9851578 TI - Novel hexarelin analogs stimulate feeding in the rat through a mechanism not involving growth hormone release. AB - Growth hormone-releasing peptides (GHRPs) are a class of small peptides that stimulate growth hormone (GH) release in several animal species, including the human. Moreover, GHRPs injected into the brain ventricles stimulate feeding in the rat. The aim of this study was to evaluate the GH-releasing properties of a series of novel GHRP analogs and the possible existence of functional correlations between the GH-releasing activity and the effects on feeding behavior. Two well-known hexapeptides, GHRP-6 and hexarelin, given s.c., dose dependently stimulated both GH release and feeding behavior in satiated rats. However, in a series of tri-, penta- and hexapeptide analogs of hexarelin, some compounds were active either on GH release or on eating behavior. Interestingly, even minor structural modifications resulted in major changes of the pharmacological profile. We conclude that GHRPs have orexigenic properties after systemic administration which are largely independent from the effects they exert on GH release. PMID- 9851580 TI - Biphasic elevation of plasma histamine induced by water immersion stress, and their sources in rats. AB - The effect of water immersion stress on the plasma concentration of histamine, in Wistar and mast cell-deficient (Ws/Ws) rats, was investigated. The histamine content of the plasma, skin and gastric mucosa, as well as the level of activity of histidine decarboxylase in the gastric mucosa, were determined by high performance liquid chromatography (HPLC)-fluorometry. In Wistar rats exposed to water immersion stress for a total of 6 h, an initial, acute, four-fold, transient increase in the plasma histamine level, followed by a sustained, though lower, elevation of the plasma histamine level, was observed. The initial acute increase in plasma histamine level was also seen in gastrectomized Wistar rats exposed to water immersion stress, but not in Ws/Ws rats exposed to stress. The sustained elevation of the plasma histamine level was observed in the Ws/Ws rats. However, in both the gastrectomized Wistar rats and gastrectomized Ws/Ws rats, the sustained elevation in plasma histamine level was not observed. The histamine content of the skin of Wistar rats after 15 min or more exposure to water immersion stress, was 20% lower than that of control rats. The mucosal histamine content of both Wistar rats and Ws/Ws rats, was 20% lower, whereas histidine decarboxylase activity in the gastric mucosa was enhanced by two-fold, during exposure to stress for 4 h. These findings indicate that water immersion stress causes a biphasic increase in plasma histamine concentration in Wistar rats; the initial acute increase in plasma histamine level originates from mast cells, and the second, sustained increase is attributed to enterochromaffin-like cells. PMID- 9851579 TI - The effect of methamphetamine on the release of acetylcholine in the rat striatum. AB - We examined the effect of methamphetamine on the release of acetylcholine in the striatum of freely moving rats, using an in vivo microdialysis method. The basal level of acetylcholine was 3.67+/-0.47 pmol/30 microl per 15 min in the presence of neostigmine (10 microM). Tetrodotoxin (1 microM), a selective blocker of voltage-dependent Na+ channels, markedly inhibited the release of acetylcholine in the striatal perfusates. Apomorphine (1.0 mg/kg, i.p.), a dopamine receptor agonist, also significantly attenuated acetylcholine release. Methamphetamine (0.1 and 0.5 mg/kg, i.p.) did not immediately affect acetylcholine release in the striatum, but a dose of 1.0 mg/kg (i.p.) induced an increase of acetylcholine release in the striatum at 15-60 min. Striatal infusion of methamphetamine (5 and 10 microM) did not influence acetylcholine release. The increase following intraperitoneal administration of methamphetamine was slightly diminished by haloperidol (0.5 mg/kg). After microinjection of the neurotoxin, 6 hydroxydopamine (6 microg/3 microl), in the substantia nigra 7 days before, the increase of acetylcholine induced by the administration of methamphetamine (1.0 mg/kg) was slightly attenuated, whereas the administration of reserpine (2 mg/kg, i.p.) 24 h before, combined with alpha-methyl-p-tyrosine (300 mg/kg, i.p.) 2.5 h before, completely blocked the increase in release of acetylcholine. These findings suggest that methamphetamine exerts an excitatory influence on striatal acetylcholine release in freely moving rats, and that this excitatory effect involves the dopaminergic system and the catecholaminergic system. PMID- 9851581 TI - Effects of water deprivation and angiotensin II intracerebroventricular administration on brain nitric oxide synthase activity. AB - Intracranial administration of L-arginine causes a reduction of the water intake induced by water deprivation or by intracerebroventricular (i.c.v.) injection of angiotensin II (angiotensin II), through the release of nitric oxide (NO) in the central nervous system. We studied the effects of i.c.v. angiotensin II (120 ng/rat) in association with i.c.v. L-arginine (2.5-10 microg/rat) on blood pressure. We also studied the effects of both peripheral and central angiotensin II injection (1.5-6 mg kg(-1) i.p. and 30-120 ng rat(-1) i.c.v., respectively) on NO synthase activity in the cortex, diencephalon and brainstem, after water deprivation (24 h), conditions producing activation of the renin-angiotensin system. L-arginine dose dependently antagonized the increase in blood pressure induced by i.c.v. angiotensin II (P < 0.001). Peripheral administration of angiotensin II produced a dose-dependent reduction of NO synthase activity in the brainstem and cortex (P < 0.001), but not in the diencephalon. Water deprivation produced similar effects on brain NO synthase activity. Angiotensin II i.c.v. injection caused NO synthase activity reduction in all brain regions studied (P < 0.001). Our findings suggest that NO and angiotensin II could play opposite roles in brain regulation of blood pressure and drinking behaviour. PMID- 9851582 TI - Effects of cytisine on hydroxyl radicals in vitro and MPTP-induced dopamine depletion in vivo. AB - The potential new iron-chelator cytisine and the radical scavenger N-tert-butyl alpha-(2-sulfophenyl) nitrone (S-PBN) were incubated in a Fenton system and hydroxyl radical formation was measured with the salicylate trapping assay. Both cytisine and S-PBN reduced hydroxyl radical formation in a concentration dependent manner. For in vivo studies, C57BL/6 mice were injected repeatedly with cytisine (0.5 mg/kg or 2.0 mg/kg s.c.) or saline seven days before and after a single 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) injection (30 mg/kg s.c.). Seven days after MPTP treatment alone dopamine levels were significantly reduced to 12% of the control values (p < 0.001), whereas MPTP + cytisine treatment (2 mg/kg) led to more than twofold higher dopamine levels (p < 0.01) compared with MPTP alone. We have shown for the first time that cytisine attenuates hydroxyl radical formation in vitro and reduces MPTP-induced dopamine depletion. Thus, cytisine may be useful for the treatment of Parkinson's Disease where the chelation of iron ions could prevent neuronal cell death. PMID- 9851583 TI - Altered neuropeptide Y Y1 responses in mesenteric arteries in rats with congestive heart failure. AB - The aim of the present study was to elucidate if the potentiating effect of neuropeptide Y on various vasoactive agents in vitro is (1) altered in mesenteric arteries from rats with congestive heart failure and (2) mediated by the neuropeptide Y Y1 receptor. The direct vascular effects of neuropeptide Y and its modulating effects on the contractions induced by endothelin-1-, noradrenaline-, 5-hydroxytryptamine (5-HT)-, U46619-(9,11-dideoxy-11alpha, 9alpha-epoxymethano prostaglandin F2alpha) and ATP, and acetylcholine-induced dilatations were studied in the presence and absence of the neuropeptide Y Y1 antagonist, BIBP3226 (BIBP3226?(R)-N2-(diphenylacetyl)-N-[(4-hydroxyphenyl)methyl ]-D-arginine amide?). Neuropeptide Y, per se, had no vasoactive effect in the arteries. The potency of endothelin-1 was significantly decreased in congestive heart failure rats. Neuropeptide Y and neuropeptide Y-(13-36) potentiated the endothelin-1 induced contraction in congestive heart failure mesenteric arteries. In 20% of the congestive heart failure rats, sarafotoxin 6c induced a contraction of 31+/ 4%. Neuropeptide Y also potentiated U46619- and noradrenaline-induced contractions but not 5-HT-induced contractions in congestive heart failure arteries. In sham-operated animals neuropeptide Y potentiated noradrenaline- and 5-HT-induced contractions. These potentiations were inhibited by BIBP3226. Acetylcholine induced an equipotent relaxation in both groups which was unaffected by neuropeptide Y. In conclusion, neuropeptide Y responses are altered in congestive heart failure rats. The potentiating effect differs between vasoactive substances. Neuropeptide Y Y1 and non-neuropeptide Y1 receptors are involved. PMID- 9851585 TI - Tyrosine kinase inhibitors suppress prostaglandin F2alpha-induced phosphoinositide hydrolysis, Ca2+ elevation and contraction in iris sphincter smooth muscle. AB - We investigated the effects of the protein tyrosine kinase inhibitors, genistein, tyrphostin 47, and herbimycin on prostaglandin F2alpha- and carbachol-induced inositol-1,4,5-trisphosphate (IP3) production, [Ca2+]i mobilization and contraction in cat iris sphincter smooth muscle. Prostaglandin F2alpha and carbachol induced contraction in a concentration-dependent manner with EC50 values of 0.92 x 10(-9) and 1.75 x 10(-8) M, respectively. The protein tyrosine kinase inhibitors blocked the stimulatory effects of prostaglandin F2alpha, but not those evoked by carbachol, on IP3 accumulation, [Ca2+]i mobilization and contraction, suggesting involvement of protein tyrosine kinase activity in the physiological actions of the prostaglandin. Daidzein and tyrphostin A, inactive negative control compounds for genistein and tyrphostin 47, respectively, were without effect. Latanoprost, a prostaglandin F2alpha analog used as an antiglaucoma drug, induced contraction and this effect was blocked by genistein. Genistein (10 microM) markedly reduced (by 67%) prostaglandin F2alpha-stimulated increase in [Ca2+]i but had little effect on that of carbachol in cat iris sphincter smooth muscle cells. Vanadate, a potent inhibitor of protein tyrosine phosphatase, induced a slow gradual muscle contraction in a concentration dependent manner with an EC50 of 82 microM and increased IP3 generation in a concentration-dependent manner with an EC50 of 90 microM. The effects of vanadate were abolished by genistein (10 microM). Wortmannin, a myosin light chain kinase inhibitor, reduced prostaglandin F2alpha- and carbachol-induced contraction, suggesting that the involvement of protein tyrosine kinase activity may lie upstream of the increases in [Ca2+]i evoked by prostaglandin F2alpha. Further studies aimed at elucidating the role of protein tyrosine kinase activity in the coupling mechanism between prostaglandin F2alpha receptor activation and increases in intracellular Ca2+ mobilization and identifying the tyrosine phosphorylated substrates will provide important information about the role of protein tyrosine kinase in the mechanism of smooth muscle contraction, as well as about the mechanism of the intraocular pressure lowering effect of the prostaglandin in glaucoma patients. PMID- 9851584 TI - Acetylcholine and tachykinin receptor antagonists attenuate wood smoke-induced bronchoconstriction in guinea pigs. AB - To study the mechanisms of wood smoke-induced bronchoconstriction, we measured total lung resistance (RL) and dynamic lung compliance (Cdyn) in anesthetized and mechanically ventilated guinea pigs. Airway exposure to various doses of wood smoke (lauan wood; 5, 10, and 15 breaths) resulted in a dose-dependent increase in RL and decrease in Cdyn. The smoke-induced changes in RL and Cdyn were significantly attenuated by pretreatment with atropine, CP-96,345 [(2S,3S)-cis-2 (diphenylmethyl)-N-((2-methoxyphenyl)-methyl)-1-aza bicyclo(2.2.2.)-octan-3 amine; a tachykinin NK1 receptor antagonist], and SR-48,968 [(S)-N-methyl-N(4-(4 acetylamino-4-phenylpiperidino)-2-(3,4-dichlorophen yl)-butyl)benzamide; a tachykinin NK2 receptor antagonist] in combination, atropine alone, and SR-48,968 alone, but were not significantly affected by pretreatment with the inactive enantiomers of CP-96,345 and SR-48,968, CP-96,345 alone, indomethacin (a cyclooxygenase inhibitor), and MK-571 [((3-(3-(2-(7-chloro-2 quinolinyl)ethenyl)phenyl((3-dimethyl amino-3-oxo-propyl)thio)methyl)propanoic acid; a leukotriene D4 receptor antagonist]. The activity of airway neutral endopeptidase, a major enzyme for tachykinin degradation, was not significantly influenced by wood smoke during the development of bronchoconstriction. We conclude that: (1) both cholinergic mechanisms and endogenous tachykinins, but not cyclooxygenase products or leukotriene D4, play an important role in the acute bronchoconstriction induced by wood smoke, and (2) the contribution of tachykinins to this airway response is primarily mediated via the activation of tachykinin NK2 receptors, but is not associated with inactivation of the airway neutral endopeptidase. PMID- 9851587 TI - Effects of Ca2+ channel blockers on Ca2+ loading induced by metabolic inhibition and hyperkalemia in cardiomyocytes. AB - The effects of the L-type (nifedipine and verapamil) and the T-type (mibefradil) Ca2+ channel blockers on the increase in intracellular Ca2+ concentration ([Ca2+]i) induced by NaCN metabolic inhibition and hyperkalemia were examined in chicken cardiomyocytes using fluorescence imaging with Fura-2. NaCN induced a slow and sustained rise in [Ca2+]i, which was not affected by pretreating the cells for 5 min with nifedipine, verapamil, or mibefradil at 100 nM or 10 microM. Pretreatment of the cells with 10 microM nifedipine, verapamil, or mibefradil for 5 min remarkably inhibited the K+-induced increase in [Ca2+]i. These inhibitory effects diminished after 48-h pretreatment with nifedipine or verapamil but not with mibefradil. Ryanodine also induces an increase in [Ca2+]i, and this effect was enhanced by 48-h pretreatment of the cells with 10 microM verapamil but not with 10 microM mibefradil. We conclude that the NaCN-induced increase in [Ca2+]i is independent of the Ca2+ influx though the L-type or T-type Ca2+ channels. Chronic inhibition of the L-type Ca2+ channels but not T-type channels may enhance the ryanodine receptor-mediated Ca2+ release, which may be responsible for the development of tolerance to their inhibitory effects on K+-induced increase in [Ca2+]i. PMID- 9851586 TI - Pharmacological analysis of protein kinases responsible for chemotaxis of rat peritoneal neutrophils. AB - Several types of kinase inhibitors were used to investigate the possible signaling pathways leading to the chemotaxis of rat peritoneal neutrophils toward macrophage inflammatory protein-2, cytokine-induced neutrophil chemoattractant-1, and platelet-activating factor. The chemotaxis and shape changes induced by each of these chemoattractants were strongly inhibited by a tyrosine kinase inhibitor (herbimycin A) and protein kinase C inhibitors (H-7 (1-(5-isoquinolinesulphonyl) 2-methylpiperazine dihydrochloride) and calphostin C). The formation of phosphatidyl 3,4,5-triphosphate in chemoattractant-stimulated neutrophils was completely inhibited by 100 nM of wortmannin, an inhibitor of phosphatidylinositol 3-kinase, whereas the chemotaxis toward each of these chemoattractants was partially inhibited (50% inhibition). The mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK-1) inhibitor PD 98059 did not inhibit the neutrophil chemotaxis. These findings suggest that the activation of tyrosine kinase and protein kinase C strongly participates in neutrophil chemotaxis and that the activation of phosphatidylinositol 3-kinase is partially involved, but that the activation of mitogen-activated protein kinase is not involved in neutrophil chemotaxis. The cross-linking of the signaling pathways for chemotaxis toward each chemoattractant was also examined. PMID- 9851588 TI - Inhibition of nitric oxide formation and superoxide generation during reduction of LY83583 by neuronal nitric oxide synthase. AB - 6-Anilino-5,8-quinolinedione (LY83583) has been widely used as an agent to reduce levels of nitric oxide (NO)-dependent cGMP in tissues. We report here that suppression of NO formation and production of superoxide during enzymatic reduction of LY83583 by neuronal NO synthase appeared to be potentially involved in the pharmacological action caused by LY83583. LY83583 suppressed neuronal NO synthase activity of 20,000 x g rat cerebellar supernatant preparation in a concentration-dependent manner (IC50 value = 12.9 microM). A kinetic study revealed that LY83583 is a competitive inhibitor with respect to NADPH, with a Ki value of 2.57 microM. With purified neuronal NO synthase it was found that LY83583 was a potent inhibitor of NO formation by the enzyme and served as efficient substrate for reduction with a specific activity of 173 nmol of NADPH oxidized per mg of protein per minute. The reductase activity was stimulated about 19.8-fold by addition of CaCl2/calmodulin, indicating that the presence of CaCl2/calmodulin is essential to express maximal activity of LY83583 reduction. Although LY83583 was a good substrate for both NADPH-cytochrome P450 reductase (P450 reductase) and DT-diaphorase, these flavin enzymes-catalyzed reductions of LY83583 were less than the neuronal NO synthase-mediated reduction in the presence of CaCl2/calmodulin. Enzymatic generation of superoxide during reduction of LY83583 by neuronal NO synthase, P450 reductase or DT-diaphorase was confirmed by electron spin resonance (ESR) experiments. Thus the present results indicate that a benzoquinone derivative LY83583 appears to interact with the P450 reductase domain on neuronal NO synthase, resulting in inhibition of NO formation and superoxide generation, which is involved in suppression of intracellular cGMP content. PMID- 9851589 TI - Receptor binding characteristics of [3H]NAD-299, a new selective 5-HT1A receptor antagonist. AB - In vitro receptor binding properties of the novel tritiated 5-hydroxytryptamine1A (5-HT1A) receptor antagonist (R)-3-N,N-dicyclobutylamino-8-fluoro-[6-3H]-3,4 dihydro-2H-1-benzo pyran-5-carboxamide ([3H]NAD-299, generic name robalzotan) were evaluated and compared with those of the agonist 8-hydroxy-2-[2,3-3H]di-n (propylamino)tetralin ([3H]8-OH-DPAT). [3H]NAD-299 binding displayed a Kd value of 0.17 nM and a Bmax value of 26.7 pmol/g wet weight of rat hippocampus. Same binding affinity (Kd = 0.16 nM) was found to cloned human 5-HT1A receptors. Addition of the nonhydrolyzable GTP analog guanylylimidodiphosphate had no effect on the binding characteristics of [3H]NAD-299, while it significantly decreased both the affinity and density of receptors labeled with [3H]8-OH-DPAT. The rank order of potency of various compounds to inhibit [3H]NAD-299 binding is consistent with the labeling of 5-HT1A receptors. This newly developed high affinity and selective antagonist radioligand provides a valuable tool for studies of 5-HT1A receptors both in vitro and in vivo. PMID- 9851590 TI - Pharmacological properties of homomeric and heteromeric GluR1o and GluR3o receptors. AB - Homomeric and heteromeric alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor subunits GluR1o and GluR3o were expressed in Spodoptera frugiperda (Sf9) insect cells. Membranes containing the recombinant receptors showed a doublet of bands of the expected size (99-109 kDa) after western immunoblotting which was shifted to a single band upon deglycosylation. In (R,S) [3H]AMPA binding experiments, high expression was seen (Bmax = 0.8-3.8 pmol/mg protein) along with high affinity binding to a single site (Kd, nM+/-S.D.): GluR1o, 32.5+/-2.7; GluR3o, 23.7+/-2.4; GluR1o + GluR3o, 18.1+/-2.9. The pharmacological profiles of these receptors resembled that of native rat brain AMPA receptors: AMPA analogues > L-glutamate > quinoxaline-2,3-diones > kainate. In the Xenopus oocyte expression system we had previously shown that the agonist (R,S)-2-amino-3-(3-carboxy-5-methyl-4-isoxazolyl)propionate (ACPA) exhibited an 11-fold selectivity for GluR3o vs. GluR1o. In this study, it was found that ACPA has 3-fold higher affinity at homomeric GluR3o and heteromeric receptors than at homomeric GluR1o, suggesting that its efficacy and/or desensitisation properties are different at GluR1o vs. GluR3o. PMID- 9851592 TI - The pharmacological profile of L-glutamate transport in human NT2 neurones is consistent with excitatory amino acid transporter 2. AB - The human teratocarcinoma cell line NTera2/D1 can be differentiated to produce post-mitotic neurones (NT2-N cells) by prolonged (> 3 week) exposure to retinoic acid. In this study, we describe the characterisation of high-affinity Na+ dependent L-glutamate transport activity in post-mitotic differentiated NT2-N cells. NT2-N cells, but not the undifferentiated precursor cells, transported L glutamate in a Na+-dependent manner, as determined by equimolar replacement of Na+ with choline. L-glutamate uptake was saturable and Eadie-Hofstee transformation of the saturation data revealed a Km of 10.6+/-0.8 microM, and a maximum transport capacity (Vmax) of 100.3+/-12.3 pmol min(-1) mg(-1) protein. Pharmacological characterisation of the transport activity in NT2-N cells produced a rank order of inhibitory activity which was identical to that determined for the human excitatory amino acid transporter 2 which we have analysed in a stable mammalian cell line (Madin Darby Canine Kidney (MDCK) cells). Of particular note, L-glutamate transport by NT2-N cells was sensitive to both dihydrokainate and kainate. The expression of human excitatory amino acid transporter mRNAs was studied using reverse transcriptase polymerase chain reaction. NT2-N cells expressed transcripts for excitatory amino acid transporters 2 and 3, but not for the subtypes 1, 4 and 5. We conclude that although the mRNA expression studies suggest the presence of transcripts for both excitatory amino acid transporter 2 and 3 in NT2-N cells, the sensitivity to dihydrokainate and kainate determined in the pharmacological analysis indicates that, of the known transporter subtypes, excitatory amino acid transporter 2 contributes to the bulk of the L-glutamate transport activity present in these cells. PMID- 9851591 TI - Effects of UTP on membrane current and potential in rat aortic myocytes. AB - The electrophysiological effects of UTP on freshly isolated rat aortic myocytes were examined using the perforated patch clamp technique. Application of alpha,beta-methylene ATP (alphabeta-meATP) and UTP, putative P2X and P2Y2 or P2Y4 purinoceptor agonists, induced transient and oscillatory inward currents, respectively. Experiments with Cl- channel blockers and different external Cl- concentrations demonstrated that the oscillatory current elicited by UTP is attributable to activation of Cl- channels. The transient component elicited by (alphabeta-meATP appeared to be responsible for a non-selective cationic current. With internal application of low-molecular-weight heparin, a blocker of inositol 1,4,5-trisphosphate (InsP3), the oscillatory current elicited by UTP was abolished. The oscillatory current was activated in an all-or-none manner by UTP over the concentration range 0.1 and 1 microM and the frequency and amplitude were independent of the UTP concentration. Under current-clamp mode, UTP produced an oscillatory membrane potential. These results show that rat aortic myocytes have at least two types of P2 receptors. Activation of the P2Y receptor by UTP produces InsP3, which releases Ca2+ from the store site. The resulting increase in intracellular Ca2+ concentration causes the oscillatory Cl- current and the subsequent membrane potential changes. PMID- 9851593 TI - HA1077, a protein kinase inhibitor, inhibits calponin phosphorylation on Ser175 in porcine coronary artery. AB - Calponin is a thin filament-associated protein which has been implicated in the modulation of the contractile state of smooth muscle via its interaction with actin and inhibition of the actin-activated myosin Mg-ATPase. This inhibitory effect is alleviated by phosphorylation of calponin at Ser175 in vitro by protein kinase C. The issue of calponin phosphorylation in intact smooth muscle in response to agonists that activate protein kinase C is controversial. We have produced a monoclonal antibody that specifically recognizes calponin phosphorylated at Ser175 and used it to analyze calponin phosphorylation in porcine coronary arterial smooth muscle stimulated with prostaglandin F2alpha or phorbol 12,13-dibutylate (PDB). Calponin phosphorylation increased rapidly in response to prostaglandin F2alpha concomitant with the increase in tension. Calponin was then dephosphorylated while force was maintained. Tension development in response to PDB was significantly slower, but again calponin phosphorylation paralleled force development. In this case, calponin dephosphorylation was very slow, consistent with prolonged activation of protein kinase C. The protein kinase inhibitors, HA1077 (1-5-(isoquinoline sulfonyl) homopiperazine HCl) and HA1100 (1-hydroxy HA1077; 1-(hydroxy-5-isoquinoline sulfonyl-homopiperazine), inhibited tension development and calponin phosphorylation in a concentration-dependent manner with similar ED50 values in response to prostaglandin F2alpha and PDB. These results support physiological roles for calponin in force development in smooth muscle in response to agonists which trigger protein kinase C activation and in the latch state, i.e., force maintenance at low energy cost. Furthermore, the vasodilator effect of HA1077 and HA1100 is more likely due to inhibition of protein kinase C than of myosin light chain kinase. PMID- 9851595 TI - Induction of apoptotic DNA fragmentation by nonsteroidal anti-inflammatory drugs in cultured rat gastric mucosal cells. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to cause apoptosis in several cell lines including transformed chicken embryo fibroblasts and human colon cancer cells. We herein report the apoptotic effect of NSAIDs in a non transformed cell line derived from the rat gastric mucosa, RGMI (rat gastric mucosa cell first). 1-[p-Chlorobenzoyl]-5-methoxy-2-methylindole-3-acetic acid (indomethacin) and sodium 2-(2,6-dichloroanilino)phenylacetate (sodium diclofenac), potent and non-selective inhibitors of cyclooxygenase, were found to induce DNA fragmentation in RGM1 cells in a time- and concentration-dependent manner. The expression of mRNA for cyclooxygenase-2 was hardly detected in the intact cells but was clearly enhanced when the cells were incubated with the two NSAIDs. In contrast, the expression of mRNA for cyclooxygenase-1 was constitutive and was never affected by NSAIDs. The effect of [3,4-di(4-methoxyphenyl)-5 isoxazolyl] acetic acid (mofezolac), a potent and highly preferential inhibitor of cyclooxygenase-1, and N-[2-(cyclohexyloxy)-4-nitrophenyl]methanesulphonamide (NS-398), a selective inhibitor of cyclooxygenase-2, on DNA fragmentation and cyclooxygenase-2 mRNA expression was weak compared to the effect of indomethacin or sodium diclofenac. The DNA fragmentation induced by sodium diclofenac was hardly affected by the exogenous addition of 16,16-dimethyl prostaglandin E2 but was inhibited by caspase inhibitors such as Ac-YVAD-CHO and Ac-DEVD-CHO. The present data provide the first evidence that NSAIDs, such as indomethacin and sodium diclofenac, cause apoptotic DNA fragmentation in cultured gastric mucosal cells, and also indicate the involvement of caspases rather than the inhibition of cellular prostaglandin synthesis in the apoptotic process. PMID- 9851594 TI - Neurotensin is an antagonist of the human neurotensin NT2 receptor expressed in Chinese hamster ovary cells. AB - The human levocabastine-sensitive neurotensin NT2 receptor was cloned from a cortex cDNA library and stably expressed in Chinese hamster ovary (CHO) cells in order to study its binding and signalling characteristics. The receptor binds neurotensin as well as several other ligands already described for neurotensin NT1 receptor. It also binds levocabastine, a histamine H1 receptor antagonist that is not recognised by neurotensin NT1 receptor. Neurotensin binding to recombinant neurotensin NT2 receptor expressed in CHO cells does not elicit a biological response as determined by second messenger measurements. Levocabastine, and the peptides neuromedin N and xenin were also ineffective on neurotensin NT2 receptor activation. Experiments with the neurotensin NT1 receptor antagonists SR48692 and SR142948A, resulted in the unanticipated discovery that both molecules are potent agonists on neurotensin NT2 receptor. Both compounds, following binding to neurotensin NT2 receptor, enhance inositol phosphates (IP) formation with a subsequent [Ca2+]i mobilisation; induce arachidonic acid release; and stimulate mitogen-activated protein kinase (MAPK) activity. Interestingly, these activities are antagonised by neurotensin and levocabastine in a concentration-dependent manner. These activities suggest that the human neurotensin NT2 receptor may be of physiological importance and that a natural agonist for the receptor may exist. PMID- 9851596 TI - Muscarinic interactions of bisindolylmaleimide analogues. AB - We have used radioligand binding studies to determine the affinities of seven bisindolylmaleimide analogues, six of which are selective inhibitors of protein kinase C, at human muscarinic M1-M4 receptors. The compounds were most potent at M1 receptors, and Ro-31-8220 was the most potent analogue, with a Kd of 0.6 microM at M1 receptors. The weakest compounds, bisindolylmaleimide IV and bisindolylmaleimide V, had Kd values of 100 microM. If it is necessary to use protein kinase C inhibitors at concentrations of 10 microM or more in studies involving muscarinic receptors then bisindolylmaleimide IV may be the most appropriate inhibitor to use. PMID- 9851597 TI - Integrating microbial decontamination with organic acids in HACCP programmes for muscle foods: prospects and controversies. AB - A considerable literature reports the antibacterial efficacy of dilute solutions of organic acids (lactic, acetic). With carcasses an overall reduction in surface contaminants of 1.5 log cycles can be expected. Carcass decontamination may not improve the safety of the resultant meat, but laboratory trials confirm that acid decontamination of subprimal and retail cuts is more efficacious. An advantage over many other intervention strategies is that residual antimicrobial activity is demonstrable over extended periods of storage. These studies have also shown that some meatborne pathogens are particularly sensitive to organic acids (i.e., Yersinia enterocolitica) while others are resistant (i.e., E. coli O157:H7). Dilute solutions of organic acids (1 to 3%) are generally without effect on the desirable sensory properties of meat when used as a carcass decontaminant. However, dependent on treatment conditions, lactic and acetic acid can produce adverse sensory changes when applied directly to meat cuts, with irreversible changes in appearance being a frequent occurrence. It is speculated that organic acid decontamination will be implemented in American abattoirs in an effort to meet specified performance standards for pathogen reduction as part of an overall HACCP program. In contrast, the EU advocates that strictly controlled processing hygiene is sufficient to ensure the safety of the product. Additional research is necessary to establish a set of treatment conditions that may permit a practicable reduction in bacterial contamination throughout the processing chain with a measurable effect on safety and storage life, without imposing any change in sensory properties. It will also be necessary to develop standard, objective measures to assess HACCP and the efficacy of decontamination procedures. Without such commercial studies controversy on the practicality of acid decontamination will persist. PMID- 9851598 TI - Nonacid meat decontamination technologies: model studies and commercial applications. AB - Increased consumer awareness and concern about microbial foodborne diseases has resulted in intensified efforts to reduce contamination of raw meat, as evidenced by new meat and poultry inspection regulations being implemented in the United States. In addition to requiring operation of meat and poultry slaughtering and processing plants under the principles of the hazard analysis critical control point (HACCP) system, the new regulations have established microbiological testing criteria for Escherichia coli and Salmonella, as a means of evaluating plant performance. These developments have renewed and intensified interest in the development and commercial application of meat and poultry decontamination procedures. Technologies developed and evaluated for decontamination include live animal cleaning/washing, chemical dehairing, carcass knife-trimming to remove physical contaminants, steam/hot water-vacuuming for spot cleaning/decontamination of carcasses, spray washing/rinsing of carcasses with water of low or high pressures and temperatures or chemical solutions, and exposure of carcass sides to pressurized steam. Under appropriate conditions, the technologies applied to carcasses may reduce mean microbiological counts by approximately one-three log colony forming units (cfu)/cm2, and some of them have been approved and are employed in commercial applications (i.e., steam-vacuuming; carcass spray-washing with water, chlorine, organic acid or trisodium phosphate solutions; hot water deluging/spraying/rinsing, and pressurized steam). The contribution of these decontamination technologies to the enhancement of food safety will be determined over the long term, as surveillance data on microbial foodborne illness are collected. This review examines carcass decontamination technologies, other than organic acids, with emphasis placed on recent advances and commercial applications. PMID- 9851599 TI - Irradiation as a method for decontaminating food. A review. AB - Despite substantial efforts in avoidance of contamination, an upward trend in the number of outbreaks of foodborne illnesses caused by nonsporeforming pathogenic bacteria are reported in many countries. Good hygienic practices can reduce the level of contamination but the most important pathogens cannot presently be eliminated from most farms nor is it possible to eliminate them by primary processing, particularly from those foods which are sold raw. Several decontamination methods exist but the most versatile treatment among them is the processing with ionizing radiation. Decontamination of food by ionizing radiation is a safe, efficient, environmentally clean and energy efficient process. Irradiation is particularly valuable as an endproduct decontamination procedure. Radiation treatment at doses of 2-7 kGy--depending on condition of irradiation and the food--can effectively eliminate potentially pathogenic nonsporeforming bacteria including both long-time recognized pathogens such as Salmonella and Staphylococcus aureus as well as emerging or "new" pathogens such as Campylobacter, Listeria monocytogenes or Escherichia coli O157:H7 from suspected food products without affecting sensory, nutritional and technical qualities. Candidates of radiation decontamination are mainly poultry and red meat, egg products, and fishery products. It is a unique feature of radiation decontamination that it can also be performed when the food is in a frozen state. With today's demand for high-quality convenience foods, irradiation in combination with other processes holds a promise for enhancing the safety of many minimally processed foods. Radiation decontamination of dry ingredients, herbs and enzyme preparations with doses of 3-10 kGy proved to be a viable alternative to fumigation with microbicidal gases. Radiation treatment at doses of 0.15-0.7 kGy under specific conditions appears to be feasible also for control of many foodborne parasites, thereby making infested foods safe for human consumption. Microorganisms surviving low- and medium-dose radiation treatment are more sensitive to environmental stresses or subsequent food processing treatments than the microflora of unirradiated products. Radiation treatment is an emerging technology in an increasing number of countries and more-and-more clearances on radiation decontaminated foods are issued or expected to be granted in the near future. PMID- 9851600 TI - Salmonella spp. on pork at cutting plants and at the retail level and the influence of particular risk factors. AB - This article describes the contamination of pork with Salmonella spp. in cutting plants and butchers' shops in The Netherlands and quantifies the influence of several risk factors. When contaminated carcasses are being processed, the main risk factors regarding cross contamination are inapt cleaning and disinfection (OR 12.8), manipulation of contaminated materials as such (OR 4.7) and (re)contaminated surfaces (OR 4.4). However, in the current situation, where contaminated carcasses are constantly being brought into cutting lines, interim cleaning and disinfection of surfaces and utensils during breaks and at the end of the working day will most likely prevent not more than about 10% of all cross contamination that takes place during a working day. Thus, as long as contaminated carcasses are being processed, about 90% of the cross contamination that occurs in cutting plants is practically unavoidable. It can therefore also be concluded that under these circumstances the implementation of codes of good manufacturing practices (GMP) and Hazard Analysis Critical Control Point (HACCP) inspired production methods will only be marginally effective in the control of Salmonella spp. cross contamination in cutting lines. The same is more or less true for the processing of contaminated cuts or carcasses by butchers in shops and supermarkets. Furthermore, in contrast to the situation in cutting plants, it may be that up to 10% of butcher's shops or kitchens of restaurants become colonized for several weeks or months with their own endemic 'house flora' of Salmonella spp., which are originally introduced via the purchased contaminated products of animal origin. Though there are no hard data to substantiate this, it can be suspected that these shops and restaurants represent the more badly managed, i.e. poorly cleaned and disinfected, enterprises. However, several analytical limitations hinder an exact determination of the prevalence of Salmonella spp. contaminated pork and an exact quantification the influence of risk factors. The diagnostic value (i.e. the sensitivity, specificity, precision and predictive value) of the combination of swabbing of carcasses and cuts and the usually employed culturing methods, in particular, is largely unknown, and there are indications that it may be seriously questioned. Without a more thorough knowledge about the diagnostic value of current and future methods of sampling and identification, it is impossible to provide for more accurate estimations of the prevalence of Salmonella positive carcasses and cuts. Based on the research data, the incidence of contaminated cuts and retail-ready pork can not be estimated more precise than as somewhere between 5-40%. When compensating for the discussed methodological flaws, it must be assumed that currently the true prevalence of contaminated primal cuts and retail-ready pork in butchers' shops is about 25-30%, and that of minced pork and pork sausages about 50-55%. Lastly it is concluded that if carcasses were Salmonella-free, consumers could in principle be provided with virtually Salmonella-free pork. It is therefore recommended that the EU allows for a decontamination step in slaughterhouses with a substance that is generally recognized as safe, provided that the producers strictly adhere to GMP-principles. PMID- 9851601 TI - Impact on human health of Salmonella spp. on pork in The Netherlands and the anticipated effects of some currently proposed control strategies. AB - The impact on human health of Salmonella spp. on pork in The Netherlands is described. Subsequently, the effects of some currently proposed control strategies in the Dutch pork production chain are evaluated and quantified with the aid of a simple mathematical model. The estimated average incidence of cases of salmonellosis in the Netherlands is about 450 cases per 100,000 person years at risk (pyar). Some special risk groups for which the risks could be quantified are (1) persons with underlying diseases, such as neoplasms or diabetes mellitus (1200 cases/100,000 pyar); (2) persons with achlorhydria or who excessively use antacids (1100 cases/100,000 pyar); (3) persons who have recently been treated with antibiotics that disturb the normal gut flora (1700 cases/100,000 pyar); (4) nurses (900 cases/100,000 pyar); (5) caterers (900 cases/100,000 pyar); (6) slaughterline personnel (1800 cases/100,000 pyar). Furthermore, it is estimated that 15% (5-25%) of all cases of salmonellosis in The Netherlands are associated with the consumption of pork. Currently, proposed control measures regarding Salmonella in pigs and on pork in The Netherlands are codes of good manufacturing practices (GMP) that, in fact, formalize recommendations that can be found in many handbooks about pig breeding and pig slaughtering. When evaluated by a mathematical model constructed for this purpose, the proposed GMP codes from farm to cutting/retail could, at best, reduce the current levels of Salmonella positive pigs and pork by 50-60%. If pigs were bred according to the rather costly specific pathogen-free concept (SPF), the prevalence of contaminated carcasses and pork could in total be reduced by 95% or more. However, implementing GMP codes from the transport phase up to the cutting/retail phase coupled with a decontamination step at the end of the slaughterline would be just as effective as GMP in combination with breeding using the SPF-concept. It is therefore concluded that the most efficient and cost-effective way of reducing the 'Salmonella problem' entailed by the consumption of pork would be to decontaminate carcasses under the precondition that the entire production chain strictly adheres to GMP principles. Therefore, the EU should also allow for more possibilities regarding the decontamination of carcasses than is currently the case. It is also concluded that current EU regulations relying on hazard analysis of critical control points (HACCP)-inspired production in cutting plants will not be effective in reducing the prevalence of Salmonella spp. on pork. This is mainly because (1) there is currently an almost steady stream of Salmonella positive carcasses that enter the cutting process; (2) when contaminated carcasses are being processed, further cross-contamination during working hours is unavoidable; (3) no steps in the cutting process are intentionally designed to effectively reduce the risks or consequences of cross contamination of cuts and retail-ready products. PMID- 9851602 TI - Host nuclear abnormalities and depletion of nuclear antigens induced in Trichinella spiralis-infected muscle cells by the anthelmintic mebendazole. AB - Infection by the parasitic nematode Trichinella spiralis induces cell cycle repositioning (chronic suspension in apparent G2/M) and genetic reprogramming in differentiated mammalian skeletal muscle cells. These changes occur in association with dramatic enlargement of infected host cell nuclei (as large as 17 microm in diameter) and nucleoli. Nuclear antigens (NA) that colocalize with host chromatin have been detected by antibodies to T. spiralis antigens, but the functions of these NA are unresolved. Mebendazole (MBZ) preferentially binds parasite versus host beta-tubulins, is implicated in inhibiting secretion in nematodes and induces cytoplasmic changes in muscle cells infected with T. spiralis. These infected cell changes might be indirect via MBZ inhibition of parasite secretions. This effect would have implications for host/parasite interactions and was evaluated here. MBZ treatment of chronically infected mice caused: (1) a significant deformation of host nuclei and diminution of nucleoli by 4 and 6 days of treatment (dot), respectively; (2) a reduction of nuclear lamins A/C in infected cell nuclei that was concomitant with nuclear deformation; and (3) significant reductions in total RNA, general protein and acid phosphatase activity levels. These changes were associated with the depletion of NA from host nuclei detected by 4 dot. However, DNA content of infected cell nuclei was not detectably reduced and muscle gene expression was not reactivated. The cellular changes documented are likely to account for previously described cytoplasmic alterations induced by MBZ. Concomitant depletion of NA from infected cell nuclei suggests a role of these products in regulating nuclear functions of host cells. PMID- 9851603 TI - Characterization of Sm20.8, a member of a family of schistosome tegumental antigens. AB - Two cDNA clones each encoding a 20.8-kDa protein (Sm20.8) were identified from the human blood fluke Schistosoma mansoni sporocyst and adult worm cDNA expression libraries by antibodies derived from rabbits vaccinated with irradiated cercariae and purified over an NP-40 extract of 3h schistosomula. Each identified cDNA has an open reading frame encoding a protein of 181 amino acids and shows homology (29-30%) with Sm21.7, Sm22.6, and Sj22.6, previously identified as belonging to a family of soluble schistosome tegumental antigens. An EF-hand calcium-binding motif is found in Sm20.8 protein in two different positions. However, neither motif binds 45calcium (45Ca) Recombinant Sm20.8 showed immunoreactivity with sera from infected humans and rabbits vaccinated with irradiated cercariae. Polyclonal rabbit sera against the Sm20.8 recognized the native protein in an extract of infected snail (sporocyst), cercariae, 3 hour schistosomules (3 h NP-40) and an adult worm preparation but not in uninfected snail tissue or eggs. Further demonstration that Sm20.8 was expressed in the different developmental stages of the parasite was by RT-PCR. Confocal microscopy demonstrates that Sm20.8 localizes to the tegument of adult worms and 3 h np-40. The IgG fraction specific to Sm20.8 mediated complement killing of schistosomules in vitro by 34%. Vaccination of mice with naked DNA containing the Sm20.8 gene and subsequently challenged with cercariae showed 30% reduction in worm burden compared to controls. PMID- 9851604 TI - Plasmodium yoelii YM MAEBL protein is coexpressed and colocalizes with rhoptry proteins. AB - We have previously cloned genes from multiple rodent malaria species exhibiting characteristics of the genes encoding Duffy binding like-erythrocyte binding proteins (DBL-EBP). Homology is seen in the intron/exon structure of the genes and in the carboxyl terminal region (including the deduced carboxyl cysteine-rich domain) of the proteins they encode. However, the amino termini of these proteins are not homologous to the DBL-EBP but contain tandem cysteine-rich regions that are similar to the cysteine-rich region of AMA-1 (apical membrane antigen-1), a rhoptry protein. This new family of proteins has been termed MAEBL and these are paralogues of both AMA-1 and the DBL-EBP. Serum against the carboxyl cysteine rich region of the Plasmodium yoelii YM MAEBL reacted to parasites with a punctate fluorescence pattern characteristic of apical organelle proteins and also localized MAEBL to the surface of merozoites within schizonts. This antiserum immunoprecipitated a protein doublet (120/128 kDa) that was unexpectedly insoluble when compared to members of the DBL-EBP. Characterization of MAEBL was extended through colocalization studies comparing the P. yoelii YM MAEBL to other parasite proteins. This protein appeared to be located in the rhoptry organelles as it colocalized with both AMA-1 and the P. yoelii 235 kDa rhoptry proteins within parasites. In addition, MAEBL is expressed relatively early in schizont development and appears on the merozoite surface after segmentation. Both the pattern and time of expression of the P. yoelii YM MAEBL are consistent with a rhoptry rather than a microneme protein. PMID- 9851605 TI - Processing of Toxoplasma ROP1 protein in nascent rhoptries. AB - Secretion in the obligate intracellular parasite, Toxoplasma gondii, occurs through a number of regulated compartments. Among these are the apical organelles known as rhoptries which release their contents as part of the invasion process. We are interested in the processing, targeting and ultimate function of rhoptry proteins (and have focused our analyses on rhoptry protein 1 (ROP1). In this paper, we address the issue of processing: using a number of engineered forms of the ROP1 gene (introduced into a ROP1- background), we show that ROP1 is synthesized as a pre-pro-protein that is subject to proteolytic cleavages to remove the pre-sequence and the 'pro' region, at the N-terminus. Using brefeldin A (BFA) and reduced temperature we show that this processing occurs late in the secretory pathway of the parasite. Immunolocalization studies with epitope-tagged constructs indicate that processing is apparently occurring in the nascent rhoptries of dividing parasites. The results are discussed in the context of the targeting and possible function of the ROP1 protein. PMID- 9851606 TI - Ether lipids and their possible physiological function in adult Schistosoma mansoni. AB - Schistosomes have lost the capability to synthesize fatty acids de novo, but they can modify fatty acids by chain elongation. This has a profound effect on the molecular species composition of the two main phospholipid fractions of schistosomes, phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Molecular species of phospholipids are increasingly recognized as important mediators, or precursors thereof, in signal transduction, immune response modulation, and events like membrane fusion. As these are all important aspects of schistosome membranes and of the tegumental membranes in particular, we analysed the PE and PC molecular species of the tegumental membranes, the worm body and the blood of the host. With the aid of on-line mass spectrometry, we unequivocally identified a large number of PC and PE species in schistosomes, among which considerable amounts of plasmalogen species. This was unexpected, as this lipid subclass has been assumed to be absent in the parasite. Species, like (20:1-16:0) diacyl PC and (16:0-20:1) plasmalogen PE, found to be main constituents in schistosomes, were absent from the blood of the host. Large differences were also found between the molecular species composition of the tegumental membranes and the membranes of the worm body. In the tegumental membranes, 1-hexadecyl 2-palmitoyl PC was detected, which could possibly function as a precursor for platelet activating factor (PAF). PMID- 9851607 TI - Cloning of a trans-spliced glyceraldehyde-3-phosphate-dehydrogenase gene from the potato cyst nematode Globodera rostochiensis and expression of its putative promoter region in Caenorhabditis elegans. AB - Reverse genetics to determine the relative importance of individual pathogenicity factors of the potato cyst nematode Globodera rostochiensis depends, apart from an efficient transformation protocol for this obligatory plant parasite, on the availability of an efficient promoter. PCR-based cloning was used to isolate a cDNA encoding glyceraldehyde-3-phosphate-dehydrogenase (GAPDH, a crucial enzyme in glycolysis and gluconeogenesis; this gene was designated gpd) and its 5' flanking region. The cDNA includes 1047 nucleotides encoding an open reading frame that shows high homology with GAPDHs from Caenorhabditis elegans and other species. Analysis of the 745 bp 5'-flanking region of the gpd gene showed no homology with a similar region in C. elegans. In this region several eukaryotic promoter elements are present. 5' Rapid amplification of cDNA ends revealed this gene was trans-spliced with a SL1 spliced leader. The 5'-flanking region of the gpd gene was fused to green fluorescent protein reporter gene and microinjected into the gonads of C. elegans. Green fluorescent protein expression, under the transcriptional control of the 5'-flanking region of gpd, was mainly observed in body wall muscles of transgenic animals. This putative promoter region of GAPDH could be a valuable tool to drive gene expression in transgenic G. rostochiensis and other related plant-parasitic nematode species. PMID- 9851608 TI - Gene structure, activity and localization of a catalase from intracellular bacteria in Onchocerca volvulus. AB - Within the context of studies on the antioxidant enzymes in Onchocerca volvulus, DNA clones encoding catalase (CAT) were isolated from an O. volvulus adult lambda zapII cDNA library. Analysis of their nucleotide and encoded amino acid sequences revealed that they derive from intracellular bacteria, rather than the O. volvulus nuclear genome. The endobacterial CAT gene was found to lie in a gene cluster, followed by a ferritin gene and an excinuclease gene. The endobacterial CAT gene encodes a functional enzyme capable of detoxifying H2O2, demonstrated by producing an active recombinant protein in an E. coli expression system. The purified 54 kDa protein has CAT activity over a broad pH range, with a specific activity of 103,000 +/- 3000 U mg(-1). The optical spectrum of the endobacterial CAT shows that it is a ferric haem-containing protein with a Soret band at 405 nm. To investigate the phylogeny of the intracellular bacterium in O. volvulus, a segment of the 16S rRNA gene was amplified from total genomic DNA by a polymerase chain reaction using universal eubacterial primers. A phylogenetic analysis of the O. volvulus-derived 16S rRNA sequence revealed that the endobacterium belongs to a distinct Wolbachia clade of the order Rickettsiales. Onchocercomata and biopsies containing different onchocercal species were immunohistochemically stained using polyclonal antibodies raised against the recombinant endobacterial CAT. CAT was detected in the endobacteria in the hypodermis of adult male and female O. volvulus, O. ochengi, O. gibsoni and O. fasciata. The endobacterial enzyme was also detected in onchocercal oocytes and all embryonic stages including intrauterine microfilariae as well as skin microfilariae. O. volvulus thus harbours Wolbachia-like endosymbionts which are transovarially transmitted and show particular affinity for the hypodermal tissues of the lateral chords. PMID- 9851609 TI - Isolation and partial characterization of a broad specificity aminotransferase from Leishmania mexicana promastigotes. AB - A broad specificity aminotransferase (BSAT), with high activity with both, aromatic amino acids and aspartate as substrates, was purified to homogeneity from promastigotes of Leishmania mexicana by a method involving chromatography on DEAE-cellulose, Red-120-Sepharose and Mono Q, and gel filtration on Sephacryl S 200. The purified enzyme showed a single band in SDS-polyacrylamide gel electrophoresis, with an apparent molecular mass of 45 kDa. Since the apparent molecular mass of the native enzyme, determined by gel filtration, was 90 kDa, the native enzyme is a dimer of similar subunits. The amino acid composition was determined, as well as the sequence of four internal peptides obtained by tryptic digestion. Two of these peptides, consisting of 49 amino acid residues in total, showed high similarity (57%) with corresponding sequences of plant aspartate aminotransferases, whereas they had only 33% identity with the aromatic aminotransferase of Escherichia coli, and 16% identity with the tyrosine aminotransferase from the related parasite Trypanosoma cruzi. The BSAT contained only one 1/2 Cys residue per monomer. The optimal pH for the enzyme reaction, with tyrosine and alpha-oxoglutarate as substrates, was 7.0. The apparent Km values for tyrosine, phenylalanine, tryptophan and glutamate, with oxaloacetate as co-substrate, were 1.3, 0.9, 0.9 and 171.8 mM, respectively; the value for aspartate with alpha-oxoglutarate as co-substrate was 2.5 mM, and that for alanine with alpha-oxoglutarate as co-substrate was 216 mM. The values for pyruvate, alpha-oxoglutarate and oxaloacetate, with tyrosine as co-substrate, were 5.6, 0.71 and 0.12 mM, respectively. These results suggest that the enzyme is a broad-specificity aminotransferase, able to transaminate the aromatic amino acids, aspartate, and to a lower extent alanine, with high sequence similarity to aspartate aminotransferases. PMID- 9851610 TI - A novel multi-domain mucin-like glycoprotein of Cryptosporidium parvum mediates invasion. AB - Cryptosporidium parvum is a protozoan parasite which produces self-limited disease in immunocompetent hosts and devastating, persistent diarrhea in immunocompromised individuals. There is no effective treatment for cryptosporidiosis and little is known about the basic biology of the organism. Cloning and sequence analysis of the gene encoding GP900, a previously identified > 900 kDa glycoprotein, predicts a mucin-like glycoprotein composed of distal cysteine-rich domains separated by polythreonine domains and a large membrane proximal N-glycosylated core region. A trinucleotide repeat composed predominantly of the triplet ACA encodes the threonine domains. GP900 is stored in micronemes prior to appearance on the surface of invasive forms. The concentration of native GP900 which inhibits 50% (IC50) of invasion in vitro is low picomolar; the IC50 for a recombinant cysteine rich-domain is low nanomolar. These observations indicate that GP900 is a parasite ligand for a host receptor involved in attachment/invasion and suggest that immunotherapy or chemotherapy directed against GP900 may be feasible. PMID- 9851611 TI - Cloning, expression and reconstitution of the trypanothione-dependent peroxidase system of Crithidia fasciculata. AB - As a consequence of aerobic metabolism, trypanosomatids are exposed to reactive oxygen intermediates such as superoxide, hydrogen peroxide and the hydroxyl radical. Metabolism of hydrogen peroxide in Crithidia fasciculata is accomplished by three distinct proteins, tryparedoxin, tryparedoxin peroxidase and trypanothione reductase, working in concert with the substrates NADPH and trypanothione. Here, we report the cloning and characterisation of the tryparedoxin (TryX) and tryparedoxin peroxidase (TryP) genes from C. fasciculata. Both genes are multicopy and organized in distinct tandem arrays in the genome. TryX encodes a 16 kDa protein, which belongs to the thioredoxin superfamily, sharing the WCPPC motif, whereas TryP encodes a 21 kDa protein belonging to a new class of peroxidases called 2-Cys peroxidoxins. Both TryX and TryP were expressed in Escherichia coli and the purified recombinant proteins shown to utilise hydrogen peroxide in the presence of NADPH, trypanothione and trypanothione reductase, similar to the native proteins. TryX is rapidly reduced by trypanothione, but weakly by glutathionylspermidine, glutathione or ovothiol A. TryP shows a broad substrate specificity and can reduced hydrogen peroxide, t butyl hydroperoxide and cumene hydroperoxide with equal efficiency. PMID- 9851612 TI - Identification and characterisation of a functional peroxidoxin from Leishmania major. AB - Leishmania spp. encounter damaging oxygen metabolites from endogenous metabolic processes as well as from exogenous sources, such as inside the gut of the sandfly vector and within host macrophages. The recently described peroxidoxin protein family form part of a novel pathway for metabolising hydrogen peroxide that, in trypanosomatids, links peroxide reduction to NADPH oxidation via trypanothione. Here we report the cloning and characterisation of the Leishmania major peroxidoxin gene, tryparedoxin peroxidase (TryP). TryP is a multi-copy gene arranged in a complex tandem array located on the size polymorphic homologues of chromosome 15. Northern analysis showed that TryP expresses a single 1.6 kb mRNA throughout promastigote development. TryP encodes a 22-kDa protein with two conserved cysteine-containing domains that defines it as a 2-Cys peroxidoxin. Purified recombinant TryP protein catabolised hydrogen peroxide in the presence of the tryparedoxin homologue from Crithidia fasciculata (Cf-TryX), trypanothione, trypanothione reductase and NADPH. The demonstration that L. major utilises a three-protein peroxidase system confirms that this is a mechanism of protection against oxidative damage in this parasite. PMID- 9851613 TI - Stage-specific activity of the Leishmania major CRK3 kinase and functional rescue of a Schizosaccharomyces pombe cdc2 mutant. AB - Cell cycle control by cdc2-related kinases (CRKs) is essential to the regulation of cell proliferation and developmental processes in many organisms. Alternating phases of growth, arrest, and differentiation are characteristics of the infectious cycle of many trypanosomatid parasites, raising the possibility that members of the trypanosomatid CRK gene family participate in the regulation of these essential processes. Here we describe properties of the CRK3 gene from Leishmania major, which encodes a 36 kDa protein kinase showing 60% amino acid sequence identity with human CDK2, including several conserved sites implicated in regulation of kinase activity. CRK3 mRNA was constitutively expressed throughout the parasite life cycle, but histone H1 kinase activity of an epitope tagged CRK3 protein was greater in log-phase than in stationary-phase promastigotes. When integrated into the genome and expressed at the optimal level, CRK3 was able to rescue the growth defect of a Schizosaccharomyces pombe cdc2 mutant (cdc2-33(ts)), indicating that CRK3 is a functional homolog of cdc2. Mutants of CRK3 at several key regulatory residues showed the expected dominant negative effects on the S. pombe mutant. This is the first example of functional expression of a trypanosomatid CRK in yeast, opening the way for further genetic studies within this amenable organism. PMID- 9851614 TI - Protein synthesis in Giardia lamblia may involve interaction between a downstream box (DB) in mRNA and an anti-DB in the 16S-like ribosomal RNA. AB - Giardia lamblia, a parasitic protozoan, has been regarded as one of the most conserved eukaryotes evolved from the prokaryotes. One of its unique features appears to be the unusually short 5'-untranslated regions (UTR) (1-6 nucleotides (nts)) and the apparent absence of 5'-cap structures from its mRNAs. Transfection of the Giardia trophozoites with luciferase-encoding chimeric transcripts, flanked by the 5'- and 3'-ends of giardiavirus (GLV) (+)-strand RNA, indicated that the translational efficiency was enhanced by 5000-fold when the 5'-viral sequence extended 264 nts into the capsid coding region and fused with the luciferase open reading frame (ORF). A 13-nt downstream box (DB) was identified within this region which complements a 15-nt sequence between nts # 1382 and 1396 near the 3'-end of the Giardia 16S-like ribosomal RNA (the anti-DB). Deletion or scrambling of this DB in the mRNA leads to a significant loss of the translational efficiency in Giardia. A Shine-Dalgarno (SD)-like element was also identified at 9-14 nts upstream from the initiation codon in the viral (+)-strand RNA, but alteration of its sequence led to no change in translation. Using the sequence complementary to ribosomal anti-DB to probe the Giardia mRNAs available in the databases, each mRNA was found to contain a putative DB with an average length from 8 to 13 nts. It is thus possible that initiation of translation in Giardia may involve a DB in the coding region of mRNA that may bind to a putative anti-DB in the small ribosomal RNA through base pairing. This mechanism of ribosome recruitment, which finds a potential parallel in Escherichia coli, could illustrate a relatively close distance between Giardia and prokaryotes in terms of translation initiation, and may provide a model for studying the evolution of translation machinery. PMID- 9851616 TI - Characterization of an Onchocerca volvulus L3-specific larval antigen, Ov-ALT-1. PMID- 9851615 TI - Overexpression of superoxide dismutase in Trypanosoma cruzi results in increased sensitivity to the trypanocidal agents gentian violet and benznidazole. AB - The parasitic protozoan Trypanosoma cruzi is exposed to toxic oxygen metabolites which arise from drug metabolism or immune mechanisms, in addition to those produced by endogenous processes. Identification and functional analysis of parasite enzymes which confer protection against oxidative stress is therefore of importance. To investigate the role of T. cruzi superoxide dismutase (SOD) we transfected epimastigotes with an expression vector containing a putative Fe-SOD gene homologue and achieved overexpression of enzyme activity (5-8 fold). Inhibition studies carried out on the partially purified enzyme revealed azide and H2O2 sensitivity and cyanide insensitivity, the profile expected of an Fe isoform. Phenotypic analysis of transformed parasites showed that they were more susceptible than control cells to growth inhibition by the trypanocidal drug benznidazole and by gentian violet, an agent which can be used to decontaminate blood supplies in endemic areas. These results may reflect an imbalance in the antioxidant defences of the parasite produced as a result of overexpression of Fe SOD. PMID- 9851617 TI - Molecular cloning of the glycosomal malate dehydrogenase of Trypanosoma brucei. PMID- 9851618 TI - Increased urinary excretion of bilirubin metabolites in association with hyperbilirubinemia after esophagectomy. AB - This study was conducted to investigate the rationale for postoperative hyperbilirubinemia after major surgery. The serum bilirubin values and urinary excretion of bilirubin metabolites (BM) were monitored in 11 patients who underwent esophagectomy via right thoracotomy for esophageal cancer. Both the serum bilirubin values and the urinary excretion of BM increased postoperatively in all patients. The maximum serum bilirubin level in four patients with septic complications, two of whom developed pneumonia and two, anastomotic leakage (group A), was significantly higher at 5.25+/-4.16 mg/dl than in the remaining patients without septic complications (group B), at 2.11+/-0.07 mg/dl. The peak value of urinary BM was 99.5+/-88.2 micromol/day in group A and 23.5 +/-26.7 micromol/day in group B. The correlation between the level of serum bilirubin and urinary BM excretion was found to be significant. Thus, the metabolism of bilirubin was increased by extensive surgical stress and septic insult. PMID- 9851619 TI - Changes and results of surgical strategies for hepatocellular carcinoma: results of a 15-year study on 452 consecutive patients. AB - In an attempt to define better surgical strategies for patients with hepatocellular carcinoma (HCC), we conducted a retrospective analysis of 452 patients who underwent hepatic resection at our institute during a period of 15 years. The patients were divided into two groups: group A, comprising 188 patients who underwent hepatic resection before 1988, and group B, comprising 264 patients after 1989. These groups were compared clinicopathologically. The percentage of patients with Child's A disease but without cirrhosis, in group A was lower. The diameter of the resected tumor was larger in group A, and major hepatic resections and curative operations were more frequently performed in group A. In group B, there was less blood loss, the specimen weighed less, and the hospital mortality was lower. Although the tumor-free survival rates were similar between the two groups, the survival rate in group B was significantly better. While even minor hepatic resection accompanied by a lower rate of surgical margin-free surgery has contributed to making hepatic resection safer, it has not improved the tumor-free survival rate. Conversely, recent advances in imaging modalities used during follow-up for the early detection of recurrence and for planning multimodality treatment have contributed to increasing the survival rate. PMID- 9851620 TI - Complications and management of microwave coagulation therapy for primary and metastatic liver tumors. AB - Microwave coagulation therapy (MCT) has been widely used, both percutaneously and directly, as effective minimal invasive therapy for liver tumors. To facilitate the use of MCT, we describe the complications we have encountered, and their possible management and prophylaxis. MCT was performed for 42 patients with hepatocellular carcinoma (HCC) and for 29 with metastatic liver tumors, following which complications developed in 14.2% and 20.6% of the HCC and metastatic groups, respectively. The complications included abscess, biloma, bleeding, hepatic failure, and dissemination of cancer cells. In the HCC group, the mean value of tumor size and the clinical stage of patients with complications were significantly larger (P = 0.006) and higher (P = 0.032), respectively, than those of patients without complications. The incidence of complications increased significantly when the tumor size was more than 4cm (P = 0.008). Abscesses and bleeding were successfully treated using percutaneous drainage and interventional angiography, respectively, but as the other serious complications were not able to be treated effectively once induced, prophylaxis is important to facilitate MCT. Transcatheter cooling of the intrahepatic bile duct during MCT and the administration of an anticancer agent into the abdominal cavity are recommended to prevent biloma and dissemination, respectively. MCT is indicated for tumors less than 4 cm in diameter to reduce the risk of complications. The prophylaxis and treatment of these complications enhance the safety of MCT. PMID- 9851621 TI - Allograft rejection of small bowel transplantation in pigs. AB - Small bowel allograft rejection in large animals has yet to be well defined. There are no specific early signs of graft rejection. The present experiments were undertaken to compare acute small bowel allograft rejection in pigs with and without FK506 and also to examine the usefulness of mucosal biopsies. Thirty-six outbred Large-White pigs were divided into (1) group 1 (n = 9): nonimmunosuppressed recipients; (2) group 2 (n = 8): FK506-immunosuppressed recipients; (3) group 3 (n = 2): autotransplant controls; and (4) donors (n = 17). Orthotopic small bowel transplantations were performed with Thiry-Vella loops for daily biopsies. The survival rate of group 2 was significantly longer than that of group 1 (P < 0.05). One best survivor in group 2 was killed at postoperative day (POD) 365. Treatment by FK506 prevented rejection, but most of the pigs died of pneumonia. In group 1, rejection began on POD 3 and progressed to severe rejection rapidly within 7 days. In group 2, rejection began from POD 6 to POD 8, but either remained mild or spontaneously improved. The differences in the routine laboratory data and the tumor necrosis factor-alpha level were not evident between the groups. Histological studies of repeated graft biopsies are thus considered to be essential for detecting signs of graft rejection. PMID- 9851622 TI - Beneficial effects of prostaglandin E1 on ischemic colitis following surgery on the abdominal aorta. AB - To determine the effects of prostaglandin E1 (PGE1) on the intestinal circulation, an experimental dog model of ischemic colitis following abdominal aortic reconstruction was made by ligating the inferior mesenteric artery (IMA) and the internal iliac artery, with the creation of a 50% stenosis in the superior mesenteric artery (SMA). The parameters of enteric ischemia included the blood flow of the SMA, the tissue blood flow in the small intestine and left colon, the mean stump pressure of the IMA (IMAP), and the IMAP/mean systemic blood pressure ratio. With the continuous infusion of 10 ng/kg per minute of PGE1 into the descending thoracic aorta, these values increased significantly on the seventh postoperative day compared with those on the day of operation. These results thus suggest that the continuous intraarterial infusion of PGE1 may reduce ischemic changes in the colon following abdominal aortic reconstruction. PMID- 9851623 TI - Effects of dynamic cardiomyoplasty on left ventricular systolic and diastolic performance in an acute canine model. AB - Although dynamic cardiomyoplasty (DCMP) is currently being evaluated as an alternative to end-stage congestive heart failure, the overall results of DCMP are variable and inconclusive. We evaluated the effect of classic DCMP on systolic and diastolic cardiac function in normal heart using reliable indicators which minimize the influences of load conditions. Six experimental dogs were evaluated with the acute nonpreconditioning model. The slope of the linear preload recruitable stroke work relationship (Mw) showed a significant increase with latissimus dorsi muscle (LDM) stimulation (postwrap non-stimulation 59.1+/ 6.3, postwrap stimulation 98.6+/-9.7 erg cm(-3) x 10(3); P < 0.01), and the x intercept (V0) was unchanged; these were utilized as the indicators of left ventricular systolic function. The constant of pressure decay (tau) increased after LDM wrap (prewrap 45.8+/-6.0, postwrap nonstimulation 69.3+/-10.3, postwrap stimulation 72.3+/-13.9 ms; P < 0.05), and the peak filling rate was unchanged after LDM wrap, which were utilized as the indicators of diastolic function. We concluded that classic dynamic cardiomyoplasty is effective in assisting systolic cardiac function, but may to some degree have a detrimental effect on the diastolic cardiac function. PMID- 9851624 TI - Esophageal intramural metastasis from an adenocarcinoma of the gastric cardia: report of a case. AB - We report herein the rare case of a 65-year-old man found to have esophageal intramural metastasis from cancer of the gastric cardia. Endoscopic examination initially revealed an infiltrating ulcerative tumor of the gastric cardia involving the esophagogastric junction, as well as a submucosal tumor of the lower esophagus. A total thoracic esophagogastrectomy with lower mediastinal lymphadenectomy was performed, and the resected specimen demonstrated that both the cardia and esophageal tumors were adenocarcinomas with the same cellular differentiation. As lymphatic invasion and metastases to the paracardial and mediastinal lymph nodes were observed, the esophageal submucosal tumor was considered to be an intramural metastasis from the carcinoma of the gastric cardia resulting from extensive lymphatic spread. The patient died of recurrent disease 9 months after the resection. This case report serves to demonstrate that intramural metastasis may be a local indicator of the systemic spread of disease in patients with gastric carcinoma, as it is in esophageal carcinoma. PMID- 9851625 TI - Complete response of esophageal cancer achieved by combination therapy with 5 fluorouracil, low-dose cisplatin, and radiation: report of a case. AB - To improve the survival rate of patients with esophageal cancer, several protocols of a preoperative combination of chemotherapy and radiotherapy, known as chemoradiation therapy, have been developed, recently characterized by the combination of 5-fluorouracil (5-FU), cisplatin, and radiation. Although some of these combinations have been demonstrated to be effective, the optimal chemoradiation dose and schedule are not yet precisely established. Recent investigations have elucidated that the radiosensitizing effects of cisplatin are able to be achieved more effectively by the daily administration of cisplatin before each fraction of radiation. Based on these investigations, we report herein the case of a patient with esophageal cancer with direct invasion to the trachea, in whom a complete response was achieved by the continuous administration of 5-FU, 600 mg/m2 per day, from days 1-5 combined with the daily administration of low-dose cisplatin, 10 mg/m2 per day before each fraction of radiation, given as 2Gy each time, throughout the entire treatment period of 3 weeks beginning on day 1. The benefits of our preoperative chemoradiation therapy included no severe side effects, down-staging and resectability of the tumor, as well as a pathological complete response, which could prolong the survival time. Our experience of this case prompts us to recommend the concurrent daily preoperative chemoradiation therapy for patients with locally advanced esophageal cancer. PMID- 9851626 TI - Signet-ring cell carcinoma of the ileum: report of a case and review of the Japanese literature. AB - We report herein the extremely unusual case of a 71-year-old woman with signet ring cell carcinoma of the ileum. She originally presented with a 6-month history of intermittent nausea and abdominal distention, but initial examinations, including gastrointestinal fiberscopy, ultrasonography, and computed tomography (CT) scan, failed to reveal any cause of her symptoms. A barium-enema study performed 11 months after her initial visit demonstrated a narrow portion of the terminal ileum. An ileocecal resection was subsequently performed, and an epigastric subcutaneous tumor was simultaneously excised. The specimen contained a tumor with a stenotic lumen resembling a "lead pipe", an ulcerative portion, and mucosa with a granular appearance adjoining its proximal site. Many small aphthous lesions with IIa + IIc appearance were seen in the apparently normal mucosa. Histopathological examination confirmed a diagnosis of signet-ring cell carcinoma. The small aphthous lesions seemed to be metastases spread via the lymphatic vessels. Our review of the medical literature revealed three cases of signet-ring cell carcinoma of the jejunum; however, this is the first reported case of signet-ring cell carcinoma of the ileum. PMID- 9851627 TI - Inflammatory pseudotumor of the abdomen: report of a case. AB - We report herein the case of a 38-year-old woman in whom a large tumor, 15 cm in diameter, was detected in the right lower quadrant of the abdomen. The tumor was removed and histological examination indicated that it was an inflammatory pseudotumor (IPT). IPT is not completely understood, but according to the few documented cases of IPT of the abdomen, its clinical behavior is benign and no recurrence has ever been reported. Thus, local resection is the most effective treatment. PMID- 9851628 TI - Early local recurrence of rectal cancer showing extremely rapid growth after curative surgery: report of a case. AB - We report herein the case of a 59-year-old woman who developed a local recurrence of rectal cancer which showed extremely rapid growth. The patient had undergone a curative low anterior resection with total mesoexcision, and was discharged on postoperative day 25 after an uneventful recovery. However, 2 months after the operation, she developed bleeding from the rectum during defecation, the quantity of which gradually increased. A colonoscopy performed during the fifth postoperative month revealed a circular tumor at the suture line. The tumor was unresectable because it had firmly invaded not only the sacrum, but also the right ureter. Despite the administration of 5-fluorouracil and leucovorin, the patient died of cancer 18 months after her initial surgery. Considering that local recurrence of rectal cancer does not usually occur within 1 year after surgery, this case is unusual because the local recurrence developed very early and showed extremely rapid growth, occupying the entire lumen of the rectum by the time it was detected by colonoscopy during the fifth postoperative month. PMID- 9851629 TI - Per anal suturing of a bleeding ulcer to achieve successful hemostasis of massive hemorrhage associated with ulcerative colitis: report of two cases. AB - We report herein the cases of two patients with ulcerative colitis complicated by massive hemorrhage who were successfully treated by per anal suturing of a bleeding ulcer. A 40-year-old woman and a 22-year-old man had suffered from extensive colitis for 8 and 2 years, respectively, and both followed a similar clinical course. They were admitted under emergency conditions and, despite treatment with intravenous high-dose prednisolone and thereafter with intraarterial injections of prednisolone, suffered several attacks of massive hemorrhage with hypovolemic shock. Emergency subtotal colectomy with ileostomy and rectal mucous fistula were performed, but recurrent massive hemorrhage occurred during the early postoperative period. Bleeding from a discrete ulcer in the rectum was subsequently located, and both patients underwent per anal suturing of the ulcer. No recurrent bleeding occurred thereafter, and an ileoanal anastomosis was performed several months later. Massive hemorrhage is a rare complication of ulcerative colitis which is most often resolved by subtotal colectomy without proctectomy. Although bleeding associated with ulcerative colitis usually occurs diffusely, these case reports serve to demonstrate that massive bleeding can also result from a discrete ulcer. We advocate that this condition be treated by an endoscopic or local hemostatic procedure. PMID- 9851630 TI - Arteriovenous malformation of the rectum: report of a case. AB - We report herein the case of a 38-year-old man found to have a rectal arteriovenous malformation (AVM). The patient was admitted to our hospital for investigation of fresh anal bleeding and general malaise. Barium-enema examination showed a slightly elevated lesion in the rectum, and a selective superior rectal angiogram subsequently revealed an AVM in the peripheral region of the superior rectal artery, which was presumed to be the cause of the anal bleeding. Colonoscopic examination disclosed a submucosal tumor-like lesion in the left posterior wall of the rectum, 3cm above the anal verge. After marking the boundaries by clipping, transanal resection of the lesion was performed. Histological examination revealed an irregularly expanded arteriovenous aggregation in the submucosal layer. The patient had a favorable postoperative course, and no residual AVM was seen on a postoperative selective inferior mesenteric arteriogram. There have been no signs of recurrence in the 2 years since his operation. PMID- 9851631 TI - Laparoscopic enucleation of a pancreatic insulinoma: report of a case. AB - We report herein the case of a 48-year-old Japanese woman in whom a pancreatic insulinoma was successfully treated by laparoscopic enucleation. The patient presented after developing episodic neurohypoglycemic symptoms, and an insulinoma in the pancreatic tail, 1.0 cm in diameter, was diagnosed by the results of biochemical and radiological examinations. A laparoscopic intraoperative ultrasonogram demonstrated a solitary hypoechogenic tumor in the pancreatic tail. After the tail and body of the pancreas with the spleen were mobilized, laparoscopic enucleation was performed without any complications. The total operative time was 225min and the estimated blood loss was 20 ml. Serial blood sugar measurements demonstrated a sharp rise in blood sugar levels at the time of enucleation. The patient's postoperative course was uneventful, and she was discharged on the seventh postoperative day. She has remained well for 33 months following surgery without any hypoglycemic symptoms. PMID- 9851632 TI - Neuroendocrine carcinoma of the extrahepatic biliary tract with positive immunostaining for gastrin-releasing peptide: report of a case. AB - We report herein the first documented case of gastrin-releasing peptide-positive neuroendocrine (NE) carcinoma of the extrahepatic biliary tract. An invasive tumor measuring 2.5 x 1.5 cm was located in the confluence portion of the cystic duct in a 70-year-old Japanese man. Histologically, the tumor was found to be composed of small polygonal cells which formed a solid and trabecular structure, and the frequencies of both mitoses and small necrotic areas were dominant. The tumor cells were immunoreactive to the NE markers chromogranin-A and neuron specific enolase, as well as to carcinoembryonic antigen and gastrin-releasing peptide. Although a few cases of gastrin-releasing peptide-positive small-cell lung carcinoma have been documented, there have been no reports of gastrin releasing peptide-positive NE carcinoma occurring in the gastrointestinal tract. We consider our case not merely to be of pathological interest, but also to have clinical and therapeutic implications. PMID- 9851633 TI - Syringomatous adenoma of the nipple: report of a case. AB - We report herein a rare case of syringomatous adenoma (SA) of the nipple. Only 22 cases of SA of the nipple have been documented in the world literature, and to the best of our knowledge this is the first case to be reported in Japan. The patient was a 68-year-old woman who presented with a painful mass in the right subareolar region of over 15 years' duration. Clinical examination including needle biopsy indicated a high possibility of carcinoma; however, the final histopathological diagnosis after mastectomy proved to be SA. SA is a benign locally infiltrating neoplasm of the nipple which shares many clinicopathological features with adenoma of the nipple. Both lesions sometimes show deceptive clinicopathological findings, causing then to be easily confused with carcinomas. However, SA can be distinguished by its unique histological appearance resembling sweat duct, or syringomatous, tumors and locally infiltrating growth, which is probably responsible for its misleading clinical findings and higher rate of recurrence. Surgeons should be aware of the possibility of diagnosing this extremely rare tumor, and the appropriate treatment. PMID- 9851634 TI - The surgical approach for descending necrotizing mediastinitis: report of two cases. AB - Descending necrotizing mediastinitis (DNM) is a potentially life-threatening complication that can develop after an oropharyngeal infection. Patients with DNM are usually treated with mediastinal drainage through a cervical or transthoracic approach, as well as by the systemic administration of antibiotics. We report herein the cases of two patients we recently treated for DNM. In the first patient, the DNM was limited to the upper mediastinum, and cervicomediastinal drainage was performed. In the second patient, the mediastinitis spread below the tracheal bifurcation down into the lower mediastinum, and an abscess was drained through a thoracotomy approach. The principles of the surgical approach for DNM are discussed, with a brief review of the literature following these case reports. PMID- 9851635 TI - Double cardiac rupture after direct infarct coronary angioplasty: report of a case. AB - The occurrence of double cardiac rupture (DCR) after a successful direct infarct coronary angioplasty (DICA) is extremely uncommon. We describe herein the case of a patient who underwent delayed DICA and despite successful recanalization, subsequently suffered DCR, as a postinfarction ventricular septal defect (PIVSD) with left ventricular free wall rupture (LVFWR). Emergency surgery was performed, and the patient is now doing well more than 2 years after his operation. The possible mechanisms of DCR after delayed DICA are discussed following the presentation of this unusual case. PMID- 9851636 TI - Successful treatment of bilateral deep femoral aneurysms and multiple iliac aneurysms associated with severe aortic valve disease: report of a case. AB - We report herein the case of a patient in whom aneurysms of the bilateral deep femoral arteries (DFA) and multiple iliac aneurysms associated with severe aortic valve disease were successfully treated by a two-staged operation. The patient was a 74-year-old man who had dense calcification of the ascending aorta and aortic arch. Prior to aortic valve replacement (AVR), the aneurysms of the DFA and internal iliac arteries were resected. The terminal end of the abdominal aorta and bilateral common iliac arteries were then reconstructed with a Y graft to be used as a possible alternative arterial input route in place of the ascending aorta for extracorporeal circulation during the AVR. The inferior mesenteric artery (IMA) was well developed, and the external iliac arteries and their branches were preserved at aneurysmectomy. Postoperatively, there was no ischemia of the pelvic organs or the hip muscles. The AVR was subsequently performed 5 weeks after the first operation, and the patient was discharged after an uneventful postoperative course. PMID- 9851637 TI - Multiple urethral calculi in children: report of two cases. AB - Multiple urethral calculi are rarely found in children. We report herein two unusual cases of children found to have multiple urethral calculi. In one, the calculi formed proximal to a stricture and in the other they formed within a diverticulum. Both patients required urethrotomy for removal of the calculi. A unique cause of urethral stricture is also described in case 1. PMID- 9851638 TI - A unique method of closure for an aortocaval fistula in association with a ruptured abdominal aortic aneurysm: report of a case. AB - We report herein the case of a 78-year-old man in whom an aortocaval fistula caused by spontaneous rupture of an abdominal aortic aneurysm (AAA) was successfully treated by a unique surgical technique. The aortocaval fistula had been revealed by an aortography after the patient presented with high-output heart failure. During the operation, massive bleeding from the fistula was evident. The fistula measured 2 cm in diameter, and was located between the right posterior wall of the AAA and the inferior vena cava (IVC). Direct suturing of the defect in the IVC failed to close the fistula because the tissue around it would not hold together due to degeneration. However, the bleeding was finally able to be controlled by plugging the fistula with isolated and properly trimmed omentum packed within the excluded aneurysmal sac. Unfortunately, the patient died due to respiratory failure on the 201st postoperative day. A pathological autopsy revealed that the aortocaval fistula had been closed by fibrous tissue and that the IVC was patent. Although such a drastic operative measure to repair an aortocaval fistula has never before been reported, it could be an alternative when direct closure proves unsuccessful. PMID- 9851639 TI - Preparation of a conjugate of mitomycin C and anti-neural cell adhesion molecule monoclonal antibody for specific chemotherapy against biliary tract carcinoma. AB - To develop a specific chemotherapy modality for the perineural invasion of neural cell adhesion molecule (NCAM) positive biliary tract cancer, an anticancer drug, mitomycin C (MMC), was covalently bound to anti-NCAM monoclonal antibody (anti NCAM MoAb) to form a conjugate using the cyanogen bromide method. The substitution ratio of the conjugate determined spectrophotometrically was 3.5 (MMC mol/immunoglobulin G mol). The cytotoxic activity of the conjugate, which was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test in the growth inhibition of the neuroblastoma cell line with NCAM expression, was maintained at 65.8% to 94.5% compared with the same concentration of MMC solution. The binding activity of the conjugate to the NCAM-positive bile duct cancer was examined by immunohistochemical staining and proved to be the same level as that of free anti-NCAM MoAb. The conjugate prepared in this study therefore appeared to be a potentially useful tool for immunotargeting specific chemotherapy against biliary tract cancer with NCAM expression. PMID- 9851640 TI - Complete dissection of the lymph nodes located along and at the roots of the left gastroepiploic vessels during distal gastrectomy. AB - Achieving complete dissection of the lymph nodes located at the roots of the left gastroepiploic vessels is difficult when distal gastrectomy is the surgical goal; firstly, because the short gastric vessels must be preserved to supply blood to the remnant stomach; and also, because the roots of the left gastroepiploic vessels at the splenic hilus are obscured intraoperatively by adipose tissues. We describe herein a technique which enables complete and easy dissection of the lymph nodes located along and at the roots of the left gastroepiploic vessels. PMID- 9851641 TI - Laparoscopic orchiopexy of intraabdominal undescended testis associated with congenital diaphragmatic hernia. AB - Laparoscopy is an advantageous method for the repair of intraabdominal undescended testis since both an abdominal exploration and vascular elongation can be effectively performed by laparoscopic assistance. A 3-year-old boy and a 1 year-old boy complaining of unilateral nonpalpable left testes were observed following previous congenital diaphragmatic hernia repairs on the day of birth. In these operations, a laparoscopic working sheath was inserted through a small supraumbilical incision. On each boy, a laparoscopically intraabdominal testis was found close to the left internal inguinal ring. In addition, a 10-mm trocar port was placed in the right lower abdominal quadrant and a 5-mm port was placed in the left lower quadrant. For the purpose of orchiopexy, the left testicular vessels were isolated over their full length. The left side of the testis was pulled through the inguinal ring to an inguinal canal divided by the abdominal wall, and then was retracted into the scrotum. The internal ring was thereafter closed externally by the fascia transversalis. Both patients are doing well, with no testicular atrophy or inguinal hernia for 1 and 2 years, respectively, after the above operations. PMID- 9851642 TI - ALS/MND and the perspective of health economics. AB - In health care, choices are constantly being made about alternative uses of scarce resources, and health economics offers a framework for analysing these choices and for improving resource allocation. In cost-effectiveness analysis, the costs and consequences of alternatives are systematically measured and compared, with the objective of achieving maximum health gain with the available resources. Treatment options for patients with ALS/MND are severely limited, but riluzole has been shown to offer modest improvements in survival. However, decision-makers are likely to want convincing evidence on the cost-effectiveness of this therapy before recommending widespread adoption. Here, some initial estimates of cost-effectiveness are provided, using published effectiveness data and considering only the costs of therapy and of tracheostomy. Compared with placebo, the incremental cost per life year gained of 50 mg/day of riluzole is pound sterling 45630, and of 100 mg/day is pound sterling 44890. Increasing the estimated costs of tracheostomy reduces the cost per life year gained of 50 mg/day to pound sterling 34940. However, if quality of life during the increased period of survival is 80% of full health, the cost per quality adjusted life year gained of 50 mg/day becomes pound sterling 57040. These cost-effectiveness ratios are well in excess of the range that is normally considered to be acceptable in UK health technology assessment. However, the comparatively small number of ALS/MND patients and the lack of treatment alternatives should also be considered. Meanwhile, better information on costs is required in order to produce more precise estimates of cost-effectiveness. PMID- 9851643 TI - Diagnosis of amyotrophic lateral sclerosis. AB - This review of the differential diagnosis of amyotrophic lateral sclerosis focuses on two themes. The first is practical, how to establish the diagnosis based primarily on clinical findings buttressed by electrodiagnosis. The main considerations are multifocal motor neuropathy and cervical spondylotic myelopathy. The second theme is the relationship of motor neuron disease to other conditions, including benign fasciculation (Denny-Brown, Foley syndrome), paraneoplastic syndromes, lymphoproliferative disease, radiation damage, monomelic amyotrophy (Hirayama syndrome), as well as an association with parkinsonism, dementia and multisystem disorders of the central nervous system. PMID- 9851644 TI - Clinical neurophysiology in the diagnosis of amyotrophic lateral sclerosis: the Lambert and the El Escorial criteria. AB - For many years, the only published criteria for the electrodiagnostic (EDX) recognition of amyotrophic lateral sclerosis (ALS) were those formulated by Lambert (1957; 1969). In 1990, different EDX guidelines were incorporated in the all-inclusive diagnostic criteria formulated by a subcommittee on ALS of the World Federation of Neurology, which met in El Escorial, Spain. Unfortunately, particularly in regard to the EDX requirements, the 'El Escorial criteria' have several flaws which compromise their usefulness. These include: (1) they ignore the fact that whenever upper and lower motor neuron disorders co-exist, as they characteristically do with ALS, the motor unit potential firing pattern is controlled by the upper motor neuron lesion; (2) they markedly devalue the usefulness of detecting fasciculations and, through presumably typographical error, state that the 'absence' rather than the 'presence' of fasciculations supports the diagnosis of ALS; this view is in direct conflict with the opinions expressed by most electromyographers; (3) they contain a statement regarding how the diagnosis of ALS is confirmed by the EDX studies which is confusing and, for two of the body regions (bulbar; thoracic), unrealistic; (4) finally, many of the EDX features they listed supporting the recognition of possible LMN degeneration appear to be mislabeled, while a few features in the EDX criteria are incorrect. PMID- 9851645 TI - Creatine kinase values in amyotrophic lateral sclerosis. AB - Serum creatine kinase (CK1) values were recorded while or soon after the diagnosis of amyotrophic lateral sclerosis (ALS) was established in 140 of 217 (65%) consecutive patients seen in the Motor Neuron Disease Clinic at this center during a 5-year period. Second creatine kinase (CK2) determinations were recorded on 48 of 140 (34%) ALS patients later in the course of the disease. The mean CK1 and CK2 values (unit, times the upper limit of normal) were 1.22 (range, 0.1 to 3.0; median, 0.80) and 1.34 (range, 0.1-9.2; median, 0.80) (P=0.11, r=0.92, mean difference=28.2%). Of the 140 ALS patients, 58 (41%) (male:female ratio: 2.9:1.0) had elevated CK1 values ranging from 1.03 to 13.0 (mean, 2.21; median, 1.67). Twelve patients (8.6%) had CK1 values greater than 3.0. Mean CK1 values were significantly greater (P=0.001) in male (mean, 1.56) versus female (mean, 0.80) ALS patients, and significantly greater (P=0.009) in limb-onset (mean, 1.34) versus bulbar-onset (mean, 0.80) disease. CK1 values correlated poorly with and were not predictive of age of onset or survival. PMID- 9851646 TI - Early diagnosis of ALS/MND. AB - With the advent of a specific biological therapy for ALS there is an increasing imperative for early diagnosis. As new, more effective therapies become available, this will become more important. It is intuitively probable that early therapy will prevent disability in this otherwise relentlessly progressive disorder. The difficulties posed by this need for early diagnosis in clinical practice are discussed. PMID- 9851647 TI - Linear estimates of rates of disease progression as predictors of survival in patients with ALS entering clinical trials. AB - Maximal voluntary isometric grip and foot dorsiflexion (FD) strength and forced vital capacity (FVC) were obtained in 62 patients with ALS at or close to enrollment into two clinical trials. The agents tested did not slow disease progression. Isometric strength data were standardized, and the worse side was taken. FVC was expressed as a percentage of the predicted value (FVC%). We derived linear estimates of rates of disease progression based on the isometric myometry and FVC measures and on disease duration. Forty one patients were known to have died or to have undergone tracheostomy for ventilatory support. Probability of tracheostomy-free survival was calculated using the Kaplan-Meier method. The measured values, the linear estimates for rates of decline of these values, gender, age at onset, bulbar vs. spinal onset, height and weight were tested as risk factors within the Cox proportional hazards model, using regression techniques. When tested individually, estimates of rates of decline based on all three measures (FD, grip and FVC%) were the only statistically significant risk factors (P<0.005). Multivariate analysis resulted in a 3 variable model (chi-square=75.3, P<0.00001) in which estimated rates of decline of FD strength and of FVC%, and bulbar onset were independently significant (P<0.0001, P<0.0007 and P<0.05, respectively). We conclude that linear estimates of the rate of disease progression till enrollment into a clinical trial may be better predictors of patient survival than demographic data or discrete biologic measures. PMID- 9851648 TI - Severe progression of ALS/MND after intervertebral discectomy. AB - We observed seven patients who developed their first signs and symptoms of motor neuron disease together with signs of protrusion/prolapse of intervertebral disc. The age of the patients was between 55 and 67, of which one female and six male patients. All of them suffered from cervical spine pain or low back pain. The female patient and one male patient developed weakness in the small feet muscles as initial symptom and they complained of paresthesia along dermatomes L5S1 and of severe pain. The other five patients developed wasting of the hands muscles. They had a rather mild pain in the cervical spine and early morning paresthesia as well as severe causalgia along dermatomes C5C6 or C6C7. After the diagnosis of compressive radiculopathy in all patients, they underwent surgical treatment and very soon developed very severe progression of muscle wasting which included muscles of limbs, trunk and bulbar innervated muscles with signs and symptoms of lower and upper motor neuron lesion. Five patients died from 12 to 15 months after surgical treatment and two patients are still living. PMID- 9851649 TI - Language impairment in motor neurone disease. AB - Evidence from recent neuropsychological studies of patients with motor neurone disease (MND) has demonstrated that some patients have cognitive impairment. This challenges traditional teaching that MND is a disease with only motor symptoms. Language processing in MND patients has so far not been studied in any depth, but has only been touched upon as a part of general cognition. This study has assessed nine MND subjects on a range of standardised language assessments and compared their performance to that of nine control subjects. Although this is a small experimental sample of subjects, the results of this study indicate that language impairment is present in some patients with MND. The deficits are subtle and only exposed on formal testing. It is not possible to define the nature of the language impairment from this restricted set of data, but there was evidence of some deficits in a sub-group of MND patients on tasks involving naming, auditory comprehension of complex sentences, some semantic tests and spelling. The implications of these findings are discussed in relationship to the clinical management of patients with MND. PMID- 9851650 TI - Mood disturbances in motor neurone disease. AB - This study explores the severity of depression and anxiety experienced by patients suffering from motor neurone disease and seeks to link this to illness progression, recent life events and personality factors. The 18 patients who participated in the study were assessed for the presence of psychiatric symptoms using the Beck Depressive Inventory and the Hospital Anxiety and Depression Scale. The severity of the illness and the life events during the course of the illness were measured using the Norris scale and the Holmes and Rahe Social Readjustment Rating Scale. A personality assessment was made using the Eysenck Personality Questionnaire (EPQ). Only four out of these 18 patients (23%) in the study suffered depression and of these three (17%) were mild and one (6%) was of moderate severity. A further six patients (33%) were rated as borderline depression. Only two patients suffered from an anxiety state and both had concurrent mild depression. No correlations of depression were found with the life events or illness progression. Only the L sub-scale of the EPQ (15.6+/-4.9) exceeds values expected for the general population. Despite the severity and frequency of life events during the course of the illness, most patients do not develop a depressive illness. The reason for this unexpected result is unclear. Depression is believed to result from underactivity of the serotonergic neurotransmitter system. However, in MND this system is relatively unaffected in contrast to dementia and other neurodegenerative disorders. The patients' high L scores suggest that many may cope with their illness by widespread use of denial as a psychological defence. PMID- 9851652 TI - Corticomotorneuronal hyper-excitability in amyotrophic lateral sclerosis. AB - We have analysed how the behaviour of a voluntarily activated motor unit changes when subjected to 100-150 threshold cortical stimuli using peristimulus time histograms (PSTHs). This is a measure of the integrity of the corticomotoneuronal core innervating a single anterior horn cell. One hundred and thirty units in 29 patients with ALS and 35 units in eight age-matched normal controls were studied. PSTHs were constructed using 1-ms bins of stimulus triggered sweeps with a total analysis time of 250 ms (50 ms before and 200 ms after the stimulus). In ALS the primary peak of the PSTH was delayed in onset and prolonged in duration. The primary peak was further analysed by finer 0.2-ms bins, which showed in ALS there were more sub-components than normally occur. Additional sub-components in the PSTH primary peak implies a hyper-excitable corticomotoneuron that fires excessively. Excitability could be glutamate induced and/or due to failure of GABA inhibitory mechanisms. Some glutamate antagonsits may be therapeutic in ALS because of their anticonvulsant or GABergic properties rather than their anti glutamate properties. GABA(B) agonists might have a role in future therapeutic combined therapies for ALS. PMID- 9851651 TI - The natural history and the effects of gabapentin in amyotrophic lateral sclerosis. AB - Glutamate excitotoxicity seems to play an important role in the aetiopathogenesis and progression of Amyotrophic Lateral Sclerosis (ALS). Gabapentin is a modulator of the glutamatergic system and has been shown to prolong survival in the transgenic model of familial ALS. It has also been demonstrated to slow the decline of arm strength in human sporadic cases. The aim of our study was to assess the effects of different dosages and duration of treatment of gabapentin on the natural history and survival of ALS patients. A total of 110 patients affected by definite ALS entered the study. After a 6-12 month period of observation, patients were randomly assigned to receive oral gabapentin 500 mg/day (Group A) or 1000 mg/day (Group B) for 6 months. In addition a group of patients received gabapentin 500 mg/day for 6 months and 1000 mg/day for a further 6 months (Group C). A group of 121 patients referred to our Institute, who received only symptomatic treatment, was considered as the control group (Group D). Each patient was seen at entry and every 3 months. All average slopes were negative but the comparison of all slopes showed a trend toward a slower rate of decline of muscle strength loss in all treated groups of patients compared with the control group. The differences were statistically significant. Analysis between the pretreatment and treatment period showed a statistically significant decrease of the decline of muscle strength and Norris score during the treatment period. Survival analysis showed a significantly longer survival in treated patients of Groups B and C. Our study suggests that gabapentin may be an effective drug for ALS; hence a controlled trial involving a sufficient large number of patients is warranted. PMID- 9851653 TI - Riluzole has no acute effect on motor unit parameters in ALS. AB - Riluzole is the only drug to have been approved for the treatment of amyotrophic lateral sclerosis (ALS/MND). Its mechanism of action is complex and includes actions on NMDA and kainate receptors and modulation of voltage gated Na channels. In ALS, its effects on measurable parameters of the motor units utilising current neurophysiological techniques are unknown. In an acute randomized, double-blind, placebo-controlled, cross-over experiment, we serially assessed the effects of riluzole on motor units in muscles affected by ALS/MND using EMG. We discuss the results of our observations in the light of previous clinical trials, and their implications. PMID- 9851654 TI - Serpin=serine protease-like complexes within neurofilament conglomerates of motoneurons in amyotrophic lateral sclerosis. AB - Neurofilamentous conglomerates (NfCg), as axonal spheroids or conglomerates in motoneurons, are the histopathologic hallmarks for early stages of amyotrophic lateral sclerosis (ALS). We hypothesize that NfCg may be formed by post translational modifications of altered Nf proteins that include: (1) hyperphosphorylation, (2) glycosylation (or glycoxidation), (3) nitration, (4) ubiquitination and/or (5) crosslinking by the Ca++-dependent transglutaminase (TGase). These, as well as other changes, are predicted to be initiated or accentuated by oxidative damage. The damaged Nf proteins then activate cascades of intracellular protein degradation which include ATP-dependent ubiquitin/proteasome proteolysis. Other proteolytic systems, either Ca++ dependent or independent, may also be activated, such as serine and cysteine protease systems. These enzymes, either lysosomal or non-lysosomal may also participate in the degradation of damaged Nf proteins being balanced by their cognate inhibitors. Protein complexes formed by these protease=inhibitor systems, along with damaged Nf proteins, may accumulate within the cell bodies as neuronal inclusions, since a number of intracellular inclusions are found in motor neurons in ALS. In the current study, we investigated the involvement of serine proteases and their serpins in NfCg formation. Pairs of three serine proteases (trypsin, chymotrypsin and thrombin) and their cognate serpins (alpha1-anti-trypsin, alpha1 anti-chymotrypsin, and protease nexin I) were probed in motoneurons with their antibodies for both NfCg and inclusions. Positive immunoreactivities for all serine proteases and their cognate serpins support the contention that the imbalance of serine proteases and internalized serpins may have a role in formation of NfCg and inclusions, and hence, the pathogenesis of ALS. PMID- 9851655 TI - Motor neurone acetylcholinesterase release precedes neurotoxicity caused by systemic administration of excitatory amino acids and strychnine. AB - We have proposed that neuronal overactivation by either stimulation of excitatory receptors or hypofunction of inhibitory circuits is a cause of excessive acetylcholinesterase (AChE) release, which, in turn, can contribute to ALS/MND pathogenesis. We investigated histochemical and histopathological changes in cell populations of the mouse spinal ventral horn upon in vivo stimulation of glutamate receptors with L-aspartate (ASP, 10-50 mg/kg, intraperitoneal: i.p.), or blockade of glycine receptors with strychnine (STRY, 2 mg/kg, i.p.). ASP in P4 P13 (postnatal age in days) but not in older mice, and STRY irrespective of age, provoked rapid, striking depletions of motor neurone AChE, and appearance of AChE activity in astrocytes. This was followed by recovery of the enzyme in most motor neurones, astrocyte activation and statistically significant changes in: brain macrophage infiltration, loss of interneurones and motor neurones and neuronophagic images including rosettes of glial cells surrounding a central 'ghost-like' motor neurone. Although AChE release preceded the neuropathology found, it is not known if its uptake is a cause of glial activation. However, it has been shown that the enzyme potentiates non-N-metyl-D-aspartate receptors identical to those that mediate astrocyte activation. AChE activity produces protons and choline, possible microglial activators. These are putative routes towards long-lasting neuropathology. PMID- 9851656 TI - Expression of nitric oxide synthase in the spinal cord in amyotrophic lateral sclerosis. AB - The expression of nitric oxide synthase was studied in human postmortem cervical spinal cord from four individuals with amyotrophic lateral sclerosis (ALS) and four individuals who had died from non-neurological causes. A novel autoradiographic method employing [3H]nitro-L-arginine, a potent inhibitor of the enzyme, as the binding ligand, was used. The expression was quantified in four discrete subregions of the grey matter, and in the dorsal and ventral white matter. The maximum density of binding in the superficial dorsal, deeper dorsal horn, and intermediate subregions of the grey matter was similar in ALS and control tissue, but it was higher in the ALS tissue, in the ventral horn, and in the dorsal and ventral white matter, compared to the corresponding regions in the control tissue. Saturation analysis of the binding indicated a single population of high affinity binding sites in all subregions in the control tissue. However, in the ALS tissue, in all subregions, the presence of two populations of binding sites with different ligand binding affinities were indicated. It is possible therefore that the expression of an inducible isotype of nitric oxide synthase may contribute to the neuropathic changes seen in ALS. PMID- 9851657 TI - SR57746A: a survival factor for motor neurons in vivo. AB - SR57746A(1-[2-(naphth-2-yl)ethy]]-4-(3-trifluoromethyl phenyl)-1,2,5,6 tetrahydropyridine, hydrochloride) is a non-peptide compound which has been shown to exhibit a wide range of neurotrophic effects both in vitro and in vivo. Here we examine the ability of SR57746A on axotomized spinal motor neuron death in the developing rat spinal cord. After postnatal unilateral section of rat sciatic nerve, there was approximately a 50% survival of motor neurons in the fourth lumbar segment (L4). Intraperitoneal injection of SR57746A for consecutive 14 days rescued motor neuron death but did not preserve the motor neuron diameter both on axotomy and non-axotomy side. These results suggest that SR57746A is a survival factor for motor neurons in vivo and may serve as therapeutic agent for damaged motor neurons. PMID- 9851658 TI - Adenovirus-mediated transfer of the neurotrophin-3 gene into skeletal muscle of pmn mice: therapeutic effects and mechanisms of action. AB - Several neurotrophic factors (CNTF, BDNF, IGF-1) have been suggested for the treatment of motor neuron diseases. In ALS patients, however, the repeated subcutaneous injection of these factors as recombinant proteins is complicated by their toxicity or poor bioavailability. We have constructed an adenovirus vector coding for neurotrophin-3 (AdNT-3) allowing for stable and/or targeted delivery of NT-3 to motoneurons. The intramuscular administration of this vector was tested in the mouse mutant pmn (progressive motor neuronopathy). AdNT-3-treated pmn mice showed prolonged lifespan, improved neuromuscular function, reduced motor axonal degeneration and efficient reinnervation of muscle fibres. NT-3 protein and also adenovirus vectors, when injected into muscle, can be transported by motoneurons via retrograde axonal transport to their cell bodies in the spinal cord. Using ELISA and RT-PCR analyses in muscle, spinal cord and serum of AdNT-3-treated pmn mice, we have investigated the contribution of these processes to the observed therapeutic effects. Our results suggest that most if not all therapeutic benefit was due to the continuous systemic liberation of adenoviral NT-3. Therefore, viral gene therapy vectors auch as adenoviruses, AAVs, lentiviruses and new types of gene transfer not based on viral vectors that allow for efficient in vivo liberation of neurotrophic factors have potential for the future treatment of human motor neuron diseases. PMID- 9851659 TI - LIF (AM424), a promising growth factor for the treatment of ALS. AB - Growth factors are theoretically promising agents for ALS therapy, but have been disappointing in subcutaneous delivery due to either toxicity or lack of major efficacy. Leukaemia inhibitory factor (LIF), was named after its effect on haemopoietic cells, and belongs to a group of cytokines which includes CNTF, IL 6, CT-1, OM and IL-11. All group members use the gp130 signal transducing subunit for intracellular signalling, but show differences in biological effect. In vitro and in vivo studies on axotomy and nerve crush models demonstrate a powerful effect of LIF in the survival of both motor and sensory neurones, while reducing denervation induced muscle atrophy. Its effects in muscle also include stimulating myoblast proliferation in vitro, and up-regulation after muscle injury. LIF will also stimulate muscle regeneration in vivo when applied exogenously after injury. In published studies of both axotomy induced neuronal death and in the Wobbler mouse models LIF is active at doses of 10 microg/kg delivered systemically, well below the expected maximum tolerated dose suggested by primate safety studies. LIF is expressed in low levels by spinal cord neurones with significant up-regulation when the neurones are damaged by BOAA toxin, an excitatory amino acid associated with a form of ALS. This augments other evidence suggesting LIF is a trauma factor playing a role in the injury response of adult neuronal tissue, and may be more effective than related growth factors. Taken together, the data suggests LIF is a physiologically relevant trophic factor with implications in clinical medicine as a therapy for ALS, and a human recombinant form (AM424), entered human clinical trials during 1998. PMID- 9851660 TI - The psychological impact of MND on patients and carers. AB - Nineteen patients with Motor Neurone Disease (MND) who had been living with their partners for at least two years prior to the onset of their illness, together with their partners, completed self-report questionnaires to investigate the impact of MND on both patients and carers. Physical disability and impact of the illness on aspects of everyday functioning were related to levels of anxiety and depression in the patients; psychological coping strategies adopted depended to some extent on symptom duration. Carers also demonstrated signs of anxiety and depression, with the latter correlating with aspects of the patients' functional impairment. Perceived strain in carers over caring for the patient correlated with a loss in intimacy in their relationship, which in turn was predicted by patients' cognitive/behavioural and communication changes. Changes in patients' social performance also correlated with the extent to which carers felt that the illness was affecting other areas of their life, the extent to which their partner dominated their thoughts and the extent to which they could control their reactions when thinking about the patient. Satisfaction with formal services and the number of social groups to which carers belonged correlated with carers' self predicted future ability to cope. PMID- 9851661 TI - The European Amyotrophic Lateral Sclerosis Health Profile Study. ALS-HPS Steering Group. AB - Quality of life is increasingly regarded as an important outcome measure in the evaluation of treatment regimes. The last decade has seen an enormous growth in the application of measures designed to assess quality of life in a vast array of medical specialities. However, the use of such measures in neurology has been limited and is virtually non-existent in amyotrophic lateral sclerosis (ALS). The European ALS Health Profile Study is a longitudinal survey of patients diagnosed with ALS or motor neurone disease in which patients are asked to complete questionnaires concerning their subjective health status. Data from clinical assessments is also collected. It is intended that the information collected will provide more systematic and detailed evidence of the impact of the disease from the perspective of the patient. This paper outlines the purpose and methodology of the project. PMID- 9851662 TI - Breaking the news in amyotrophic lateral sclerosis. AB - Telling the diagnosis to patients with amyotrophic lateral sclerosis (ALS) is a daunting task for any neurologist. Obviously, breaking the news in ALS is not a standardizable procedure. However, proven techniques exist to reduce the trauma to the patient and ease the burden on the doctor, thus reducing the risk of burnout and the tendency to 'pull away' from the patient. Such communication skills are of fundamental importance to clinical practice and should be more prominent in medical teaching. The way the patient is told the diagnosis is now recognized to be the first and one of the most delicate steps in palliative care. Information is best offered in a stepwise fashion at the patient's pace with an emphasis on positive aspects, and in the presence of the patient's family. Reviewing available therapeutic options and current research efforts may foster hope for the future, while pointing out that almost all symptoms of ALS can be alleviated by palliative therapy may help to reduce fears. Encouraging patients to ask questions and disclose anxieties is important for their psychological wellbeing. Available options for mechanical ventilation should be reviewed early enough to allow for unhurried decision-making. We believe that the terminal phase of the disease should be discussed at the latest when dyspneic symptoms appear, in order to prevent unwarranted fears of 'choking to death'. PMID- 9851663 TI - Quality of life for ventilator-dependent ALS patients and their caregivers. AB - Seven ventilator-dependent ALS patients and eleven caregivers were interviewed in order to assess the impact of ventilator-dependence on patients and their families. The ALS Care Database questionnaires were administered with special attention to components derived from the Health Status Survey (SF-12) and ALS Quality-of-Life Index (ALSQLI) as well as the ALS Patient Caregiver Form. Six patients had difficulty communicating and one patient was totally unable to communicate. Patients had maximal limitation of daily activities as measured by The ALS QLI, yet a self-reported satisfactory quality-of-life. Caregivers were heavily burdened and their outside activities were severely limited. PMID- 9851664 TI - Building a care and research team. AB - The provision of high quality care for people with Motor Neurone Disease (MND) is challenging. The physical and psychological health needs experienced in this progressively disabling disease necessitate input from many disciplines. In order to integrate the delivery of care, the preferred model for MND is a disease specific team. Such teams can offer a coordinated range of skills, expertise and clinical experience in a setting of interprofessional support. Teams may adopt several formats; multidisciplinary, interdisciplinary and transdisciplinary. In multidisciplinary teams the different professions work to individually set goals and meet to discuss their progress. In interdisciplinary teams goals are first agreed by the team, whose members then coordinate their input to the common treatment plan. In transdisciplinary teams, not only goals but skills are shared. Teamwork allows the coordination of input and the sharing of skills and experience in solving complex clinical problems. Greater flexibility and integration of work make best use of resources. Disease-specific teams act as an educational resource and stimulate awareness of the condition. Teams should undertake clinical research, to strengthen their practice and demonstrate their effectiveness. Effective teamwork, whether in a clinical or research team, requires issues of responsibility, leadership, interprofessional rivalry and communication to be addressed. PMID- 9851665 TI - Service provision for patients with ALS/MND: a cost-effective multidisciplinary approach. AB - Optimal management of patients with ALS/MND requires a team approach, with early referral to paramedical services for clinical assessment and prompt intervention. As the condition progresses, a flexible approach to management must be adopted by the medical team, with an ability to intervene at very short notice. We have developed an efficient multi-disciplinary clinic that services the ALS/MND population of Ireland by combining the existing infrastructure of community services with a hospital-based specialist clinic. The clinic operates on a weekly basis, and is staffed by a core team including a neurologist, a liaison nurse, and the director of the ALS/MND Association. On-site and same-day physiotherapy, occupational therapy and speech therapy is available, as is pulmonary evaluation. All patients utilising the clinical services are automatically included on the Irish Register of Motor Neurone Disease, and are tracked by the liaison nurse. The core members of the clinic interact regularly with paramedical staff within the community, ensuring that necessary community services are made available within 1-2 weeks of the clinic visit. Equipment necessary for the patient's well being is made available free of charge by the Irish Motor Neurone Disease Association, following an appropriate request from the regional para-medical staff. We have thus demonstrated that an effective multi- disciplinary care service for ALS/MND can be developed at modest cost by close personal liaison between the existing health care structures and core members of a multidisciplinary team. PMID- 9851666 TI - DNA vaccination with glutamic acid decarboxylase (GAD) generates a strong humoral immune response in BALB/c, C57BL/6, and in diabetes-prone NOD mice. AB - The technique of DNA-based vaccination was used to generate a T-cell-dependent antibody response to glutamic acid decarboxylase (GAD) in BALB/c, C57BL/6, and non-obese diabetic (NOD) mice. Plasmids were constructed in which the expression of the rat GAD65 (rGAD65) or the rat GAD67 (rGAD67) gene was driven by the immediate early region promoter of the human cytomegalovirus (pCMV). This "naked" plasmid DNA was then injected into the regenerating muscles of the studied mice. In the vaccinated animals, antibody responses to GAD65 or to GAD67 were induced. Epitope recognition of GAD was studied by protein footprinting, a technique which makes use of a limited proteolysis of antibody-bound antigen. Different epitope recognition patterns were found, corresponding to strain-specific patterns. Mild trypsin treatment generated 50 kD, 46 kD, 40 kD, 30 kD, and 21 kD proteolytic fragments. In NOD mice, 50, 46 and 40 kD bands were the most prominent signals. In non-diabetes prone BALB/c mice, a faint 40 kD band appeared suggesting a rather weak protection of GAD from tryptic lysis. The pattern observed in C57BL/6 mice was more comparable to the NOD mice pattern with prominent 40 kD and 30 kD signals and a faint 21 kD fragment. Diabetes incidence was unchanged in NOD mice, and no diabetes was observed in C57BL/6 and BALB/c mice, respectively. The data demonstrate that genetic immunization is a suitable novel tool to stimulate and to manipulate an immune response against the diabetes-associated protein glutamic acid decarboxylase. Interestingly, our results indicate that, by genetic vaccination, distinct B-cell epitopes were generated in the various studied mouse strains. PMID- 9851667 TI - Monoclonal antibody enhanced stimulation of human growth hormone action in vitro using ovine costal cartilage growth plate chondrocytes. AB - Investigations into the mechanism underlying antibody-mediated enhancement of growth hormone action have been hampered by the lack of an in vitro assay system. In this work, we have used isolated ovine costal cartilage growth plate chondrocytes to demonstrate, for the first time, that monoclonal antibody EB1 can enhance the proliferative actions of human growth hormone on this cell type. Chondrocytes were cultured for 14 days prior to exposure to GH+/-monoclonal antibody EB1 for a 4-day treatment period. Human growth hormone alone promoted a significant dose-dependent increase in chondrocyte proliferation; maximal stimulation was achieved at about 3.3-10 ng/ml growth hormone, and at higher doses of growth hormone, the response declined. Monoclonal antibody EB1 was shown to enhance the proliferative activity of 10 ng/ml human growth hormone in a significant dose dependent fashion. In conclusion, our results demonstrate that an antibody capable of enhancing GH activity in vivo also has the capacity to potentiate GH activity in vitro. This system may provide an important tool for investigations into the mechanism of GH action, and how this is modified by GH enhancing MAbs. PMID- 9851668 TI - Effect of adrenalectomy and glucocorticoid therapy on lipid metabolism of lactating rats. AB - Although adrenal glucocorticoids were known to be important for adequate milk production, little is known about their effects on the metabolism of mammary glands during lactation. In this study, lactating Wistar rats on the 12th day of lactation were divided in the following groups: sham-operated (SO) and adrenalectomized (ADX) receiving no treatment; SO and ADX starved for 24 h and refed intragastrically with 2.5 ml of 50% glucose solution, 2 h before the experiment (SOR and ADXR) and ADX receiving substitute therapy with dexamethasone (ADX + DEX). Sacrifices were performed 2 days after surgery. Weight, lipid content and in vivo lipogenesis rate were evaluated in mammary gland (M.GLAND), liver, parametrial white adipose tissue (PARA) and interescapular brown adipose tissue (BAT). ATP citrate lyase activity was measured in M.GLAND, liver and PARA of SO, ADX and ADX + DEX. The rate of lipogenesis and 14CO2 production from 14C glucose by isolated acini from M.GLAND and plasma glucose were also determined. In ADX rats, food intake, lipid content, in vivo lipogenesis rate and ATP citrate lyase activity in M.GLAND were significantly lower than those in SO rats. The M.GLAND lipogenesis rate of SOR group was similar to the value found in SO rats. In ADXR rats, the M.GLAND lipogenesis rate was not normalized. However, the therapy with DEX elevated lipid content, in vivo lipogenesis rate and ATP citrate lyase activity to levels similar to those in SO. These results suggest that the glucocorticoids are essential for the occurrence of normal lipid synthesis in M.GLAND during lactation. PMID- 9851669 TI - Expression of insulin-like growth factor binding protein 1-6 genes in adrenocortical tumors and pheochromocytomas. AB - The insulin-like growth factor (IGF) system appears to be important in the regulation of adrenal growth and hormone synthesis. As IGF-binding proteins (IGFBPs) modify IGF bioactivity, we investigated the expression of IGFBP 1-6 genes in different adrenal tumors and hyperplasias to further clarify the role of the IGF system in adrenal pathophysiology. IGFBP-1 mRNA levels were too low to be detected by Northern blot analysis, but could be found by RT-PCR in some tumors and hyperplastic adrenals. Other IGFBPs were detected by Northern blotting. IGFBP 3 mRNA levels were very low in normal adrenals. In adrenal tumors and hyperplastic adrenals, IGFBP-3 mRNA expression was usually higher than in normal adrenals. In hormonally active adrenocortical carcinomas, IGFBP-2, -4, -5 and -6 mRNA levels were lower than in nonfunctional carcinomas and normal adrenals. The low IGFBP mRNA expression in the hormone-producing carcinomas was associated with high IGF-II mRNA content. In adrenocortical adenomas from patients with Cushing's or Conn's syndrome, mean IGFBP mRNA levels were higher than in normal adrenals or in hormonally inactive adenomas. In nodular and bilateral hyperplasias, IGFBP-2, 3 and -4 mRNA expression was on average higher than in normal adrenals but varied substantially, as did IGFBP mRNA levels in pheochromocytomas. In comparison to normal adrenals, pheochromocytomas expressed on average higher levels of IGFBP-2 and -4 but less IGFBP-5 and -6 mRNAs. Our data show that the six IGFBPs 1-6 are expressed at variable level in adrenal tumors and hyperplasias. The low level of IGFBP mRNAs in hormonally active adrenocortical carcinomas was of particular interest. PMID- 9851670 TI - Expression of transforming growth factor-beta1 and transforming growth factor beta Type-II receptor mRNA in papillary thyroid carcinoma. AB - Transforming growth factor-beta1 (TGF-beta1) is a potent inhibitor of epithelial cell proliferation. Signal transduction by TGF-beta1 involves direct binding to the TGF-beta Type-II receptor and then the formation of a heterodimeric complex of TGF-beta Type-I and Type-II receptor. To explore the role of TGF-beta1 in thyroid carcinoma, we examined the expression of TGF-beta1 and TGF-beta Type-II receptor mRNA by northern blotting analysis in both 14 papillary thyroid carcinomas and surrounding normal thyroid tissues. Relative mRNA level was determined by scanning densitometry of the autoradiogram and corrected for loading differences using a human beta-actin cDNA probe. The relative mRNA levels of TGF-beta1 in 12 out of 14 papillary thyroid carcinomas were higher than those in surrounding normal thyroid tissues. In contrast, the relative mRNA levels of TGF-beta Type-II receptor were reduced to 60.1+/-18.3% of those of normal thyroid tissues in 10 papillary thyroid carcinomas. There were no clear relationships between the relative mRNA levels for TGF-beta1 and TGF-beta Type-II receptor and the histological characteristics of papillary thyroid carcinomas. The relative mRNA levels for TGF-beta1 and TGF-beta Type-II receptor did not show significant differences in thyroid carcinomas with or without lymph node metastases. There was a negative correlation between the TGF-beta Type-II receptor mRNA level and tumor size, while no significant correlation was observed between the TGF-beta1 mRNA level and tumor size. In conclusion, most papillary thyroid carcinomas overexpress TGF-beta1 mRNA but exhibit a reduction in TGF-beta Type-II receptor mRNA. The reduction of TGF-beta Type-II receptor mRNA may play a role in the pathogenesis of papillary thyroid carcinoma. PMID- 9851671 TI - Beta3-adrenergic receptor gene polymorphism is not associated with hypertension in NIDDM patients without nephropathy. AB - OBJECTIVE: We studied the relationship between a Trp64Arg mutation of beta3 adrenergic receptor gene and hypertension in Japanese NIDDM patients without nephropathy. RESEARCH DESIGN AND METHODS: Eighty-three Japanese NIDDM patients (46 men, 37 women, known duration of 12 [5-31], age of 55 [40-69] years, median and [range]) with normoalbuminuria and microalbuminuria were studied. The Trp64Arg mutation in the beta3-adrenergic receptor gene was determined by the PCR RFLP method. RESULTS: The frequency of the mutated allele was not greater in the hypertensive patients (18.2%, n=37) than in the normotensive (20.2%, n=46) patients. Genotypes with a mutate allele (i.e. Trp64/Arg64 + Arg64/Arg64) were observed in 23.5% in the hypertensive patients and in 35.7% in the normotensive patients, respectively. A similar finding in the allele frequency was observed when the analysis was made separately in the normoalbuminuric and microalbuminuric patients, respectively. CONCLUSION: These results suggest that a Trp64Arg mutation in the beta3-adrenergic receptor gene is not associated with hypertension in Japanese NIDDM patients without nephropathy. PMID- 9851672 TI - Determination of plasma ET-1 by RIA--risks and limitations. AB - Plasma concentrations of the vasoconstrictor peptide endothelin-1 (ET-1) were determined in nonobese healthy subjects as well as in unspecified patients using three different commercially available radioimmunoassays (RIAs) and the respective recommended prepurification procedures. Depending on the antibody and extraction procedure applied different ET-1 concentrations were measured in identical plasma samples. Thus, this study clearly documents that ET-1 concentrations measured in plasma samples represent apparent concentrations which have to be interpreted with caution. PMID- 9851673 TI - Growth hormone improves weight velocity and height velocity in prepubertal children with cystic fibrosis. AB - We undertook this study to determine if growth hormone treatment of prepubertal children with cystic fibrosis could improve their height and weight. Nine prepubertal children with cystic fibrosis were treated with human recombinant growth hormone for one year. Results obtained during this year were compared to similar measurements made for each patient for the one year prior to the treatment year. Anthropometric data including: height, height velocity, weight, weight velocity and skin fold thickness were measured at three month intervals. Pulmonary function and skeletal muscle strength were measured at three month intervals. Glucose tolerance was evaluated by HbAlc and by fasting blood glucose and insulin levels every three months. Our results demonstrate that growth hormone treatment resulted in significant improvement in height velocity and height Z scores. Weight increased in all subjects, with a significant increase in weight velocity (year prior to treatment = 1.7+/-1.0 kg/yr, treatment year = 3.8+/-1.6 kg/yr; p=0.03). Measurements of skin fold thickness suggests that lean body mass improved with growth hormone treatment. Pulmonary function improved in all but two patients, whose pulmonary function remained the same and muscle strength improved in all subjects. These results suggest that growth hormone used in prepubertal children with cystic fibrosis can improve height and weight and may improve lean body mass. PMID- 9851675 TI - Relationships of serum leptin to body composition and resting energy expenditure. AB - Body weight (BW), body mass index (BMI), body fat, serum leptin concentration and resting energy expenditure (REE) were fitted in a multiple linear regression model in a group of individuals with stable body weight. While serum leptin concentration was well related to BW, to BMI and to body fat, no correlations with the REE values were found. This suggests that serum leptin concentration would represent an index of adiposity and poorly reflects energy metabolism. PMID- 9851674 TI - Effect of testosterone replacement on whole body glucose utilisation and other cardiovascular risk factors in males with idiopathic hypogonadotrophic hypogonadism. AB - BACKGROUND: Excessive testosterone in males or estrogens in females could explain their differences in coronary heart disease event rates. As a contraceptive testosterone is likely to be used at large scale the role of testosterone in increasing the risks of coronary heart disease needs investigation. AIM: To look at the role of testosterone in development of insulin resistance and other cardiovascular risk factors. DESIGN: Prospective, before-after study on ten male subjects with idiopathic hypogonadotrophic hypogonadism pre- and post testosterone replacement therapy; outcome measures: anthropometry, lipoprotein profile and M value (whole body glucose disposal rates on standard hyperinsulinemic euglycemic clamp; at insulin infusion rate: 40 mU x (m-2)). RESULTS: Pre-treatment serum testosterone was 0.43 (0.515) ng x mL(-1), LH was 1.29 (0.08) IU x L(-1), and FSH was 1.54 (0.08) IU x L(-1). None had glucose intolerance. After replacement testosterone levels increased to 9.4 ng x mL(-1) (p=0.0005); weight increase of 5.0 kg (p=0.140), body mass index increase of 1.2 kg x m(-2) (p=0.28), and the change in waist to hip ratio (p=0.31) were not statistically significant. M-value (mg x kg x min(-1)) did not change after testosterone therapy (5.86 [0.72] vs 5.29 [0.82], p=0.62). Insulin levels (mU x L(-1)) achieved during the clamps were 89.5 (14.2) before and 146 (32.2) after androgen therapy (p=0.127). There was no change in glucose area under curve (mg x min x dL(-1)) (14406 [502.2] vs 12557 [826.5], p=0.312). On testosterone replacement therapy total and LDL cholesterol levels (mg x dL(-1)) declined (122.5 [13.4] vs 91.6 [5.0], p=0.04; 65.9 [9.9] vs 39.4 [7.3], p=0.05); Ratio of total cholesterol to HDL ratio also decreased significantly (p=0.05). Changes of serum triglycerides (p=0.25) and HDL cholesterol (p=0.19) did not attain statistical significance. CONCLUSIONS: Insulin sensitivity does not decrease on testosterone replacement therapy of male subjects with idiopathic hypogonadotrophic hypogonadism. Testosterone replacement was associated with decrease in other cardiovascular risk factors. PMID- 9851676 TI - Kinetic characterization of CYP2E1 inhibition in vivo and in vitro by the chloroethylenes. AB - Trans- and cis-1,2-dichloroethylene (DCE) isomers inhibit their own metabolism in vivo by inactivation of the metabolizing enzyme, presumably the cytochrome P450 isoform, CYP2E1. In this study, we examined cytochrome P450 isoform-specific inhibition by three chloroethylenes, cis-DCE, trans-DCE, and trichloroethylene (TCE), and evaluated several kinetic mechanisms of enzyme inhibition with physiological models of inhibition. Trans-DCE was more potent than cis-DCE, and both were much more effective than TCE in inhibiting CYP2E1. The kinetics of in vitro loss of p-nitrophenol hydroxylase (pNP-OH) activity (a marker of CYP2E1) in microsomal incubations and of the in vivo gas uptake results were most consistent with a mechanism in which inhibition of the metabolizing enzyme (CYP2E1) was presumed to be related to interaction of a reactive DCE metabolite with remaining substrate-bound, active CYP2E1. The kinetics of inhibition by TCE, a weak inhibitor in vitro, were very different from that of the dichloroethylenes. With TCE, parent compound concentrations influenced enzyme loss. Trans-DCE was a more potent inhibitor of CYP2E1 than cis-DCE based on both in vivo and in vitro studies. Quantitative differences in the inhibitory properties of the 1,2-DCE isomers may be due to the different stability of epoxides formed from bioactivation by CYP2E1. Epoxide intermediates of DCE metabolism, reacting by water addition, would yield dialdehyde, a potent cross-linking reagent. PMID- 9851677 TI - Species differences in the glutathione transferase GSTT1-1 activity towards the model substrates methyl chloride and dichloromethane in liver and kidney. AB - Glutathione transferase (GST) GSTT1-1 is involved in the biotransformation of several chemicals widely used in industry, such as butadiene and dichloro methane DCM. The polymorphic hGSTT1-1 may well play a role in the development of kidney tumours after high and long-term occupational exposure against trichloroethylene. Although several studies have investigated the association of this polymorphism with malignant diseases little is known about its enzyme activity in potential extrahepatic target tissues. The known theta-specific substrates methyl chloride (MC) dichloromethane and 1,2-epoxy-3-(p-nitrophenoxy)propane (EPNP) were used to assay GSTT1-1 activity in liver and kidney of rats, mice, hamsters and humans differentiating the three phenotypes (non-conjugators, low conjugators, high conjugators) seen in humans. In addition GSTT1-1 activity towards MC and DCM was determined in human erythrocytes. No GSTT1-1 activity was found in any tissue of non-conjugators (NC). In all organs high conjugators (HC) showed twofold higher activity towards MC and DCM than low conjugators (LC). The activity in human samples towards EPNP was too close to the detection limit to differentiate between the three conjugator phenotypes. GSTT1-1 activity towards MC was two to seven-times higher in liver cytosol than in kidney cytosol. The relation for MC between species was identical in both organs: mouse > HC > rat > LC > hamster > NC. In rats, mice and hamsters GSTT1-1 activity in liver cytosol towards DCM was also two to seven-times higher than in the kidney cytosol. In humans this activity was twice as high in kidney cytosol than in liver cytosol. The relation between species was mouse > rat > HC > LC > hamster > NC for liver, but mouse > HC > LC/rat > hamster/NC for kidney cytosol. The importance to heed the specific environment at potential target sites in risk assessment is emphasized by these results. PMID- 9851678 TI - Characterization of CYP1A in hepatocytes of cynomolgus monkeys (Macaca fascicularis) and induction by different substituted polychlorinated biphenyls (PCBs). AB - Cynomolgus monkeys (Macaca fascicularis) have been used previously as a model to study effects on cytochrome P450 (CYP) regulation. Until now it has not been elucidated which CYP1A proteins are present in this primate species. The aim of this study was to characterize CYP1A in untreated hepatocytes of cynomolgus monkey using two specific CYP1A inhibitors (alpha-naphthoflavone and furafylline). The effect of different substituted polychlorinated biphenyls (PCBs) on CYP1A regulation was also studied in these hepatocytes. Small quantities of CYP1A2 have been identified in untreated hepatocytes. Northern blots showed the presence of a CYP1A mRNA in untreated hepatocytes, when hybridizations where performed with human CYP1A2 cDNA. Inhibitions with furafylline and alpha-naphthoflavone also suggested the presence of CYP1A2 properties. After induction with different PCBs, (probably) CYP1A1 mRNA and enzyme activity were induced in cynomolgus monkey hepatocytes. As expected, 2,3',4,4',5-PeCB (PCB no. 118), a mono-ortho substituted congener, was a potent CYP1A inducer but 2,2',3,4,4',5',5'-HpCB (PCB no. 180), a di-ortho and 2,2',3,4',5,5',6-HpCB (PCB no. 187), a tri-ortho substituted PCB, could induce CYP1A mRNA and enzyme activity in cynomolgus monkey hepatocytes as well. PMID- 9851679 TI - Inhibition of prostaglandin-H-synthase by o-phenylphenol and its metabolites. AB - Chronic administration of o-phenylphenol (OPP) is known to induce urinary bladder tumours in the Fischer rat. The underlying toxic mechanism is poorly understood. Recently, arachidonic acid (ARA)-dependent, prostaglandin-H-synthase (PHS) catalysed metabolic activation of the OPP metabolite phenylhydroquinone (PHQ) to a genotoxic species was suggested to be involved in OPP toxicity. To investigate this hypothesis in more detail, we have studied the effects of OPP and its metabolites on PHS. When microsomal PHS from ovine seminal vesicles (OSV) was used as enzyme source, both OPP, PHQ, and 2-phenyl-1,4-benzoquinone (PBQ) inhibited PHS-cyclooxygenase. The inhibitory potency was inversely related to the ARA concentration in the assay; at 7 microM ARA IC50-values were: 13 microM (OPP), 17 microM (PHQ), and 190 microM (PBQ). In cells cultured from OSV, which express high PHS activity, 40 microM OPP almost completely suppressed prostaglandin formation. Studies with microsomal PHS demonstrated that PHQ was an excellent substrate for PHS-peroxidase; both ARA and hydrogen peroxide supported oxidation to PBQ. OPP was only a poor substrate for PHS, but inhibited the ARA mediated and to a lesser extent also the hydrogen peroxide-mediated in vitro oxidation of PHQ. Moreover, PHQ at up to moderately cytotoxic concentrations (50 microM) did not induce micronuclei in OSV cell cultures. Taken together, our findings do not provide evidence for an ARA-dependent, PHS-catalysed formation of genotoxic species from PHQ. Moreover, it seems to be questionable whether such activation can effectively occur in vivo, since OPP and PHQ turned out to be efficient cyclooxygenase inhibitors, and high levels of OPP and PHQ were found at least in the urine of OPP-treated rats. On the other hand, inhibition of the formation of cytoprotective prostaglandins in the urogenital tract may play a crucial role in OPP-induced bladder carcinogenesis. PMID- 9851680 TI - Mutagenicity testing of organic extracts of diesel exhaust particles after spiking with polycyclic aromatic hydrocarbons (PAH). AB - In the present study, spiking was used as a strategy to evaluate the mutagenicity of individual compounds in a mixture. Mutagenicity of individual polycyclic aromatic hydrocarbons (PAH) was evaluated in an organic extract of diesel exhaust particles (DEP). The particles were extracted with dichloromethane (DCM). After replacing DCM with dimethylsulphoxide (DMSO), the extract was spiked with four individual PAH: benzo(a)pyrene, benzo(a)anthracene, pyrene and fluoroanthene. The PAH were added separately and in various combinations to the extract to determine the effects of each variable and to identify possible interactions between the individual PAH and between the PAH and the extract. The study was designed as a fractional factorial experiment with the five variables (the DEP extract and the four PAH), giving 16 (instead of 32) mixtures plus a triplicate centrepoint and background, i.e. a total of 20. The fractionated factorial design used in the present work supports a model with linear and interaction terms. The mixtures were tested for mutagenicity in the Ames assay using four strains of Salmonella typhimurium in the presence of rat liver xenobiotic enzymes (S9-mix). Projections to Latent Structures (PLS) was used to quantify the mutagenicity of each compound and possible interactions. The four individual PAH and the DEP extract acted additively in the Ames test with 10% S9-mix. PMID- 9851681 TI - Murine strain differences and the effects of zinc on cadmium concentrations in tissues after acute cadmium exposure. AB - The role of strain differences in cadmium tissue distribution was studied using sensitive (129/J) and resistant (A/J) mice. These murine strains have previously been shown to differ in their susceptibility to cadmium-induced testicular toxicity. Cadmium concentration was measured in testis, epididymis, seminal vesicle, liver, and kidney at 24 h after cadmium chloride exposure (4, 10, and 20 micromol/kg CdCl2). The 129/J mice exhibited a significant increase in cadmium concentration in testis, epididymis, and seminal vesicle at all cadmium doses used, compared to A/J mice. However, cadmium concentrations in liver and kidney were not different between the strains, at any dose, indicating that cadmium uptake is similar in these organs at 24 h. These murine strains demonstrate similar hepatic and renal cadmium uptake but significantly different cadmium accumulation in the reproductive organs at 24 h. The mechanism of the protective effect of zinc on cadmium toxicity was studied by assessing the impact of zinc acetate (ZnAc) treatment on cadmium concentrations in 129/J mice after 24 h. Zinc pretreatment (250 micromol/kg ZnAc), given 24 h prior to 20 micromol/kg CdCl2 administration, significantly decreased the amount of cadmium in the testis, epididymis, and seminal vesicle of 129/J mice, and significantly increased the cadmium content of the liver after 24 h. Cadmium levels in the kidney were unaffected at this time. Zinc pretreatment also prevented the cadmium-induced decrease in testicular sperm concentration and epididymal sperm motility seen in 129/J mice. These findings suggest that the differences in the two murine strains may be attributed partly to the differential accumulation of cadmium in murine gonads. This may be caused by strain differences in the specificity of cadmium transport mechanisms. The protective role of zinc in cadmium-induced testicular toxicity in the sensitive strain may be due to an interference in the cadmium uptake by susceptible reproductive organs. PMID- 9851682 TI - Regional selectivity to ochratoxin A, distribution and cytotoxicity in rat brain. AB - Ochratoxin A (OTA) a chlorodihydro-isocoumarin linked through an amide bond to phenylalanine, is a mycotoxin found as a contaminant in foodstuffs and shown to be nephrotoxic, teratogenic, immunosuppressive, genotoxic, mutagenic and carcinogenic in rodents. Ochratoxin A is known to induce teratogenic effects in neonates (rats and mice) exposed in utero, characterised by microcephaly and modification of the brain levels of free amino acids. Since OTA has been found to accumulate in the brain according to the duration of exposure to doses in the range of natural contamination of feedstuffs, experiments were designed to determine more precisely the structural target of OTA in the brain. After intracerebral injection, OTA (403 ng/10 microl) was not found in the following parts of the brain: the frontal cortex (FC), striatum (ST), ventral mesencephalon (VM) and the cerebellum (CB) in contrast to the rest of the brain, probably due to the detection limit of 0.1 ng/g of tissue. However lactate dehydrogenase (LDH) was increased in extracellular space in the VM to a greater extent than in the rest of the brain, indicating that this structure could be one of the targets of OTA in the brain. Contents of free amino acids were morever similarly modified in the VM and in the rest of the brain. Male rats were given OTA (289 microg/kg per 24 h) by gastric intubation for 8 days and the main brain structures analysed for OTA content and cytotoxicity. OTA was found in the following structures in decreasing order: rest of the brain (50.3%), cerebellum (34.4%), VM (5.1%), striatum (3.3%) and hippocampus (2.9%) of the total OTA amount found in the brain, which represents 0.022% to 0.028% of the given dose. Interestingly cytotoxicity as measured by lactate dehydrogenase (LDH) release in the extracellular space was much more pronounced in the VM, hippocampus, and striatum than in the cerebellum, whereas no cytotoxicity was observed in the rest of the brain. Similarly deoxyribonuclease (DNase) activity in relation to possible necrotic cells was increased in the VM and cerebellum. Altogether these results designated the ventral mesencephalon, hippocampus, striatum and cerebellum as the main OTA-targets in the brain of adult rats and excluded the rest of the brain. PMID- 9851683 TI - Similar effects of cis-diamminedichloroplatinum(II) and cis-diammine-1,1 cyclobutanedicarboxylatoplatinum(II) on sodium-coupled glucose uptake in renal brush-border membrane vesicles. AB - cis-Diamminedichloroplatinum(II) (cDDP) has been shown to interfere with reabsorption processes in renal tubular epithelia, leading to polyuria, magnesium and sodium wasting and glucosuria. cDDP inhibits the Na+-coupled uptake of methyl alpha-D-glucopyranoside (MGP) in renal proximal tubular cells in primary culture. cis-Diammine-1,1-cyclobutane dicarboxylatoplatinum(II) (CBDCA) produces tubular injury qualitatively similar to that of cDDP with a reduced severity. CBDCA inhibits Na+-coupled MGP uptake in renal proximal tubular cells in primary culture at concentrations 20- to 30-times higher than those of cDDP. The Na+/glucose cotransport protein possesses sulphydryl groups (SH) essential for its activity. Platinum complexes have strong affinity for SH groups. We compared the direct effects of cDDP (0.04-1.0 mM) and CBDCA (1-30 mM) on Na+-coupled MGP uptake in rabbit renal brush-border membrane (BBM) vesicles. cDDP and CBDCA inhibited Na+-coupled MGP uptake in a concentration-dependent manner, mainly through a decrease in Vmax of the cotransport protein. These effects were associated with platinum binding to BBM and decreases in protein-bound SH groups. CBDCA altered Na+-coupled MGP uptake at concentrations 30-times higher than those of cDDP. When BBM vesicles were preincubated with cDDP or CBDCA, diethyldithiocarbamate (an antidote against cDDP-induced nephrotoxicity) partly restored Na+-coupled MGP uptake and reduced the amount of platinum bound to BBM, but did not restore protein-bound SH groups. These findings strongly suggest that the inhibition of Na+-coupled MGP uptake by cDDP and CBDCA is mainly mediated by direct chemical binding of platinum to essential SH groups of the cotransport protein but may also involve other nucleophilic groups, such as the SCH3 group of methionine residues. PMID- 9851684 TI - Quantitative analysis of O-isopropyl methylphosphonic acid in serum samples of Japanese citizens allegedly exposed to sarin: estimation of internal dosage. AB - A convenient and rapid micro-anion exchange liquid chromatography (LC) tandem electrospray mass spectrometry (MS) procedure was developed for quantitative analysis in serum of O-isopropyl methylphosphonic acid (IMPA), the hydrolysis product of the nerve agent sarin. The mass spectrometric procedure involves negative or positive ion electrospray ionization and multiple reaction monitoring (MRM) detection. The method could be successfully applied to the analysis of serum samples from victims of the Tokyo subway attack and of an earlier incident at Matsumoto, Japan. IMPA levels ranging from 2 to 135 ng/ml were found. High levels of IMPA appear to correlate with low levels of residual butyrylcholinesterase activity in the samples and vice versa. Based on our analyses, the internal and exposure doses of the victims were estimated. In several cases, the doses appeared to be substantially higher than the assumed lethal doses in man. PMID- 9851685 TI - Cloning and sequence of guinea pig interleukin 2 (IL-2). AB - Interleukin 2 (IL-2) is a T-cell proliferation factor released from TH0- and TH1 type helper T-cells and is an essential cytokine for certain immune responses. We reported here cloning and sequence of IL-2 cDNA in guinea pigs, which have been used for a long time in various immunological experiments and in vivo screening tests for skin sensitization potential of chemicals. Consequently, a cDNA clone was obtained encoding guinea pig IL-2 of 520 bp in length, which contained a complete open reading frame. Alignment of the amino acid sequence with human IL-2 indicates that guinea pig IL-2 is composed of 20 amino acids (aa) of a signal peptide and 132 aa of a mature peptide with a predicted molecular weight of 15133. Guinea pig IL-2 has an amino acid homology of 62% with human IL-2, 52% with murine IL-2, and 55% with rat IL-2. In addition, guinea pig IL-2 has a possible N-linked glycosylation site as seen in bovine and porcine IL-2. PMID- 9851686 TI - Biochemistry of tropoelastin. PMID- 9851687 TI - Under respiratory growth conditions, Bcl-x(L) and Bcl-2 are unable to overcome yeast cell death triggered by a mutant Bax protein lacking the membrane anchor. AB - We have reported earlier that cytosolic expression of the full-length human apoptosis inducer Bax-alpha (Bax) in the yeast Saccharomyces cerevisiae suppresses growth and induces mortality in cells containing functional mitochondria. Human Bcl-x(L) overcomes this toxicity. Here we describe that a mutant Bax protein, with a missing membrane anchor region (Bax delta), also inhibits growth and causes cell death in yeast. However, the death inhibitory proteins Bcl-x(L) and Bcl-2 fail to rescue Bax delta-mediated growth inhibition under conditions promoting respiration, although they bind Bax delta in the cell. Results in Jurkat T-cells corroborate that Bcl-x(L) is much less efficient at rescuing mammalian cells from the effect of Bax delta than from full length Bax. We have also inquired if the respiration-dependent toxicity of Bax and Bax delta in yeast is nullified by Bcl-x(L)delta and Bcl-2delta, molecules which lack membrane anchors but bind Bax in the yeast two-hybrid system. It appears that, under conditions which facilitate respiration in yeast, Bcl-x(L)delta and Bcl 2delta are incapable of rescuing both Bax-containing and Bax delta-containing cells. Our results open up the interesting possibility that there might exist proteins, unrelated to the Bcl-2 family, which could negate death induced by a membrane anchor-free form of Bax. PMID- 9851688 TI - Characterization of cytochrome c-556 from the purple phototrophic bacterium Rhodobacter capsulatus as a cytochrome-c peroxidase. AB - A cytochrome c-556 was purified from Rhodobacter capsulatus and the complete amino acid sequence was determined. It contains 328 amino acid residues and two typical heme-binding sites at cysteine residues 54 and 57 and at residues 200 and 203. It is homologous to the family of bacterial cytochrome c peroxidases (BCCP) with 69% identity to Paracoccus denitrificans BCCP and 60% identity to Pseudomonas aeruginosa BCCP for which there is a three-dimensional structure. There is lesser similarity to the mauG gene products from methylotrophic bacteria which are thought to be involved in biosynthesis of the quinone cofactor of methylamine dehydrogenase. Translated genes from Escherichia coli and Helicobacter pylori are also related to the bacterial cytochrome c peroxidases. The divergence of this family of proteins is reflected in the fact that the reported sixth heme ligands are not conserved, except in Pseudomonas, Rhodobacter and Paracoccus. This suggests that homologs of BCCP may fold differently and/or may not have the same enzymatic activity as the prototypic protein from Ps. aeruginosa. We found that the Rb. capsulatus BCCP is active with both Rb. capsulatus cytochrome c2 and with horse cytochrome c as substrates (Km values 60 microm and 6 microm, respectively). The turnover number was 40 s(-1) and the Km for peroxide was 33 microm. We have thus confirmed that the Rb. capsulatus protein is a cytochrome c peroxidase. PMID- 9851689 TI - Human matrix metalloproteinase-9: activation by limited trypsin treatment and generation of monoclonal antibodies specific for the activated form. AB - For many studies on matrix metalloproteinases in immunohistochemistry it is important to be able to distinguish between the zymogen and activated forms of the enzymes. Activated human matrix metalloproteinase-9 (MMP-9, gelatinase B) was produced from the proenzyme by limited digestion with trypsin. The products of cleavage were characterised by SDS/PAGE and N-terminal sequencing. Trypsin treatment led to a stepwise removal of the propeptide domain and also caused cleavage within the C-terminal domain. Monoclonal antibodies specific for the activated form of human MMP-9 were raised by using a peptide corresponding to the N-terminus of the activated enzyme as immunogen. The antibodies do not recognise the MMP-9 proenzyme or the active or proenzyme forms of matrix metalloproteinase 2 (MMP-2, gelatinase A) and do not react with unrelated proteins in an unfractionated tissue extract. The antibodies were used to detect, by immunohistochemistry, activated MMP-9 in formalin-fixed, wax-embedded sections from a series of oesophageal cancer cases previously shown to contain MMP-9. All of the tumours contained activated MMP-9 localised to tumour cells and macrophages. As the antibodies are effective in immunohistochemistry on formalin fixed, wax-embedded sections, they should prove useful for the detection of activated MMP-9 in various disease processes. PMID- 9851690 TI - Structural analysis of the chicken hydroxyindole-O-methyltransferase gene. AB - Hydroxyindole-O-methyltransferase (HIOMT) catalyzes the final step of melatonin synthesis, a neurohormone involved in photoperiodism and produced specifically in the pineal gland and in the retina. In the chicken, HIOMT gene transcription appears to be controlled by a circadian oscillator located in the pineal gland. We have characterized the chicken HIOMT gene over 17 kb, including 2.9 kb of 5' flanking sequence. The major transcript (1.6 kb) is composed of eight exons distributed over 7.5 kb of genomic DNA. A ninth alternative exon was identified 6 kb downstream of exon 8. It was found in minor transcripts in the pineal gland and in the retina. Sequence similarity between exons 8 and 9 suggests their origin by exon duplication. Due to early stop codons, inclusion of exon 9 truncates the open reading frame by up to 33%. A restriction fragment length polymorphism was detected for a BglII site in intron 8. Fluorescence hybridization localized the HIOMT gene on chicken chromosome 1q22. The 5' flanking region contains GATTAA and TAATCC sequences that may be related to tissue-specific expression. An ATTTAAAT sequence at position -29 would play the role of a TATA box, as evidenced by electrophoretic mobility shift assay. Information obtained in this study open the way to further studies aimed at analyzing the circadian rhythm of transcription at promoter level. PMID- 9851691 TI - The gene for the Alzheimer-associated beta-amyloid-binding protein (ERAB) is differentially expressed in the testicular Leydig cells of the azoospermic by w/w(v) mouse. AB - In order to discover possible new testicular paracrine factors involved in the establishment of spermatogenesis, a modified differential display reverse transcription, polymerase chain reaction (DDRT-PCR) procedure was used to detect gene transcripts preferentially expressed in the testes of the azoospermic w/w(v) mutant mouse. One of the differentially expressed gene products showed partial similarity to members of the short-chain alcohol dehydrogenase family of enzymes. This cDNA fragment was used to obtain the full-length mouse cDNA sequence, which initially showed moderate similarity to a 20beta-steroid dehydrogenase from lower organisms, and later shown to have >85% similarity to a novel endoplasmic reticulum-associated-binding protein (ERAB) from the human brain, implicated in Alzheimer's disease. A recently cloned bovine sequence also of high similarity suggests that this molecule might also represent an isozyme of 3-hydroxyacyl-CoA dehydrogenase. Using the mouse cDNA as probe, northern hybridization showed enrichment of the transcript to the testicular Leydig cells, and showed a specific approximately 20-fold enrichment in the azoospermic mouse testis. The level of the testicular ERAB transcript does not seem to change through puberty, suggesting that a lack of germ cells alone is not responsible for the up regulation in the w/w(v) testis. Using the three-dimensional coordinates of the published 20beta-hydroxysteroid dehydrogenase structure as template, it was additionally possible to construct a molecular model of the novel protein which showed it to have a very similar structure to this enzyme, including the substrate-binding domain. PMID- 9851692 TI - Beta2-microglobulin can be refolded into a native state from ex vivo amyloid fibrils. AB - Beta2-microglobulin fibrils have been extracted from the femoral head of a patient who has been under chronic haemodialysis for 11 years. The primary structure of the N-terminal portion of the protein and mass determination by electrospray mass spectrometry demonstrate that beta2-microglobulin, extracted as fibrils by the water extraction procedure, was not glycated and that Asn17 was not deamidated. Limited proteolysis was observed in more than 20% of beta2 microglobulin molecules and the main cleavage sites were at the C-terminus of Lys6 and Tyr10. Beta2-microglobulin from fibrils has been purified by gel filtration in 6 M Gdn/HCl and submitted to a refolding procedure. The refolding conditions have been determined through the study of the unfolding pathway of the native protein. Beta2-microglobulin is stable at neutral pH where it displays a lower tendency to self-aggregate than in acidic conditions. Pulse dilution and extensive dialysis in refolding buffer at pH 7.5 yields beta2-microglobulin with a tertiary structure identical to that of the native form. The CD spectrum in the near-ultraviolet region and the spectrum of the intrinsic fluorescence of Trp overlap those of the native protein, but the CD spectrum in the far-ultraviolet region is affected by the contribution of oligomers created by beta2 microglobulin fragments that reduce the positive light polarisation at 205 nm typical of native beta2-microglobulin. PMID- 9851693 TI - Stimulation of fructose 1,6-bisphosphate production in melanoma cells by alpha melanocyte-stimulating hormone-- 31P/13C-NMR and 32P-labeling studies. AB - In a 31P-NMR spectroscopic study of cultured M2R mouse melanoma cells, we previously demonstrated the acute stimulation of three peaks in the phosphomonoester region of the spectrum by [Ahx4, DPhe7]alpha-melanotropin (concomitant with an increase in cellular adenosine 3',5'-phosphate (cAMP) and a decrease in ATP [Degani, H., DeJordy, J. O. & Salomon, Y. (1991) Proc. Natl Acad. Sci. USA 88, 1506-1510]. Chemical identification of these metabolites was performed in this study using 32P metabolic labeling and polyethyleneimine cellulose thin layer chromatography in combination with 31P-NMR and 13C-NMR spectroscopic methods. Two of the stimulated signals were identified as P1 and P6 of fructose 1,6-bisphosphate (FruP2) and their mode of regulation by alpha melanotropin was examined. The FruP2 response to alpha-melanotropin coincided in time and dose with a rise in cAMP and a decrease in levels of ATP, while elevation of cAMP by forskolin alone did not increase FruP2. The stimulatory effect of alpha-melanotropin was not associated with a change in the overall rate of glycolysis, suggesting that FruP2 levels were not rate limiting in this process. The data suggest the presence of a previously unknown response of M2R melanoma cells to alpha-melanotropin, which coincides in time with enhanced cAMP accumulation but is not mediated by cAMP and may relate to the control of FruP2 in a non glycolytic context. PMID- 9851694 TI - Corticotropin-releasing factor receptor type 1 from Tupaia belangeri--cloning, functional expression and tissue distribution. AB - A cDNA clone encoding corticotropin-releasing factor (CRF) type 1 (CRF-R1) has been isolated from the tree shrew Tupaia belangeri with a PCR-based approach. The full-length cDNA encoded a 415-amino-acid protein with highest sequence identity (approximately 98%) to human CRF-R1 and slightly less identity to rat or mouse CRF-R1 (approximately 97%). Only eight amino acids (residues 3, 4, 6, 35, 36 and 39 in the N-terminus, residue 232 in transmembrane domain 4 and residue 410 in the C-terminus) differed between tree shrew CRF-R1 (tCRF-R1) and human CRF-R1 (hCRF-R1). tCRF-R1 mRNA was detected by semiquantitative RT-PCR and RNase protection analysis in the pituitary and in brain areas such as amygdala, brainstem, cerebellum, cortex, olfactory bulb, and striatum. In peripheral organs, only weak expression of tCRF-R1 mRNA was observed in ovary, testis, and adrenal gland. Binding studies using human embryonic kidney 293 (HEK293) cells stably transfected with tCRF-R1 showed that the CRF agonists ovine CRF (KD = 1.28 nM), human/rat CRF (KD = 1.09 nM), urocortin (KD = 0.37 nM) and sauvagine (KD = 0.77 nM), respectively, were bound with significantly higher affinities than the CRF antagonist astressin (KD = 12.4 nM). In agreement with the binding data half maximum effective EC50 values of 0.83 nM (human/rat CRF), 1.41 nM (ovine CRF), 1.25 nM (rat urocortin) and 0.71 nM (sauvagine) were calculated when the cAMP production in HEK293 cells stably transfected with tCRF-R1 was stimulated with the four CRF analogues. These data underline the close relationship between human and tree shrew CRF-R1. PMID- 9851696 TI - Widespread presence in mammals and high binding specificity of a nuclear protein that recognises the single-stranded telomeric motif (CCCTAA)n. AB - We have recently identified a protein in HeLa nuclear extracts which recognises the single-stranded telomeric sequence (CCCTAA)n in vertebrates [Marsich, E., Piccini, A., Xodo, L. E. & Manzini, G. (1996) Nucleic Acids Res. 24, 4029-4033]. In this paper we provide further experimental evidence, using electrophoretic mobility shift assays, SDS/PAGE after ultraviolet cross-linking, and gel permeation chromatography techniques, that: (a) this protein displays remarkably stringent requirements for the telomeric motif sequence, as (CCCTAAA)n, (CCCCAA)n and (TCCCAA)n are tightly bound, but (CCTAA)n is not; (b) it requires at least four CCC-block repeats properly spaced to bind strongly to DNA, e.g. the polypurine stretch of the murine Ki-ras promoter d(CTCCCTCCCTCCCTCCTTCCCTCCCTCCC), the CarG-motif-containing sequence d(CCATTTCCTAATTAGGTAAAAG), and d(C)22 are not recognised by this protein; (c) it is present in nuclear extracts from several vertebrate sources including human, rat, pig, hamster and chicken; (d) its molecular mass is about 40 kDa, as determined by SDS/ PAGE and non-denaturing gel permeation chromatography, suggesting that this protein is monomeric under native conditions. PMID- 9851695 TI - Lyotropic-salt-induced changes in monomer/dimer/tetramer association equilibrium of formyltransferase from the hyperthermophilic Methanopyrus kandleri in relation to the activity and thermostability of the enzyme. AB - Formyltransferase from Methanopyrus kandleri is composed of only one type of subunits of molecular mass 32 kDa. The enzyme requires the presence of lyotropic salts for activity and thermostability. We report here that the enzyme is in a monomer/dimer/tetramer association equilibrium, the association constant being affected by lyotropic salts. At 0.01 M K2HPO4/KH2PO4, pH 7.2, the enzyme (0.4 mg/ml) was mainly present in a monomeric form. Upon increase of the phosphate concentration, the concentration of the dimer increased up to a phosphate concentration of 0.6 M, then decrease at the expense of tetramer formation up to a phosphate concentration of 1.0 M. The specific activity at 4 C increased from <0.1 U/mg at 0.01 M, over 1.5 U/mg at 0.6 M to 3.6 U/mg at 1.0 M. Similar results were obtained with ammonium sulfate as lyotropic salt. The findings indicate that both oligomerization and activity increase with increasing salt concentrations, suggesting that there is a causal connection. To determine this, we exploited the observation that oligomer formation was not induced by the weak lyotropic salt NaCl up to a concentration of 1.5 M and that the dissociation of the dimer into the monomer at 4 degrees C proceeded very slowly (50% in approximately 6 h). This allowed us to study the effect of NaCl on the activity of the oligomers at NaCl concentrations not sufficient to induce oligomerization. At 4 degrees C, the activity of the oligomers increased from 0.3 U/mg at 0.25 M NaCl to 3.4 U/mg at 1.0 M NaCl. At these NaCl concentrations, the monomers were inactive. The findings indicate that oligomerization is a prerequisite for enzyme activity in the presence of NaCl. The salt-dependent induction of oligomerization was parallelled by an increase in thermostability; strong lyotropic salts conferred thermostability at much lower concentrations than the weak lyotropic NaCl. PMID- 9851697 TI - Self-association of the C-terminal domain of the hepatitis-C virus core protein. AB - The N-terminal region of the hepatitis-C virus (HCV) core protein is rich in basic residues, while the C-terminal end of the protein comprises of a stretch of hydrophobic amino acids. Between these two extremes is an amphipathic region with two predicted alpha-helical segments. This region embodies Leu or hydrophobic residues in positions of heptad repeats and is possibly capable of self association. To investigate this possibility, the core sequence was divided into two fragments and expressed separately as recombinant proteins. Recombinant proteins with the N-terminal fragment remained as monomers even at high concentrations in SDS/PAGE. Recombinant protein with the C-terminal fragment appeared largely monomeric on denaturing gels but some oligomers were also detected. Furthermore, proline mutations in either one of the predicted alpha helices adversely affected the observed oligomerization. The self-association capacity of the core protein C-terminal region was further supported by results from a yeast two-hybrid system. To affirm our conclusion, a peptide covering the heptad repeats and the predicted alpha helices was synthesized. Data from mass spectrometry and gel-filtration chromatography concluded that this peptide readily self-associated into the homodimer. Therefore, our results suggest that the oligomerization motifs of the HCV core protein may not be limited to the previously suggested N-terminal region. PMID- 9851698 TI - Determinants of the fidelity of processing glucoamylase-lysozyme fusions by Aspergillus niger. AB - Fusion proteins are used to enhance the yields of heterologous proteins secreted from filamentous fungi. In Aspergillus niger, the target protein is normally fused downstream of the carrier protein glucoamylase with a Lys-Arg KEX2-like cleavage site at the junction. This is cleaved in vivo to release mature protein but the processing is not always accurate. We have used N-terminal mutant lysozymes to vary the sequence immediately downstream of the KEX site, and also varied the amino acid sequence upstream of the KEX processing site, to study the fidelity of processing. The sequences both upstream and downstream of the KEX2 site affected the fidelity of cleavage. With some constructs, a range of processing sites were apparent and the relative proportions were time dependent in batch cultures of A. niger. Aberrant processing was related to the secondary structure preferences of the amino acids in and around the KEX site. Downstream of the processing site, the fidelity of processing decreased in proportion to the tendency for helix formation. PMID- 9851699 TI - An SP1-like 5'-GACCACGCC-3' sequence is critical for activity of the inflammatory phospholipase A2 promoter and binds several non-zinc finger proteins. AB - We have previously shown that the promoter of the type IIa secreted phospholipase A2 gene contains a strong positive regulatory proximal element [-125 to -85] element B. Mutation of this element abolishes the activation of the phospholipase A2 promoter by C/EBPbeta in HepG2 cells. Liver nuclear proteins form three major and two minor complexes with this element. The [-107 to -99] 5'-GACCACGCC-3' sequence is critical for the formation of these complexes and the activity of the promoter. Although the sequence of element B is highly similar to those of Sp1 binding sites, it does not bind Sp1 or other zinc-finger proteins. Each major complex contains a single protein, the molecular masses of these proteins being 100, 90 and 75 kDa. These proteins have the same nucleotide requirements for binding, with the cytosines at positions -102, -100, -99 and the adenosine at 103 being the most important nucleotides. The activity of the phospholipase A2 promoter in HeLa cells was lower than in HepG2 cells, and was correlated with the absence of complex 3 in HeLa cell nuclear extracts. Our results suggest different roles for the proteins bound to the 5'-GACCACGCC-3' sequence. In particular, the 75-kDa protein which forms the third complex is critical for the activity of the promoter of the secretory phospholipase A2 gene. PMID- 9851700 TI - Transcriptional regulation of the tissue-type plasminogen-activator gene in human endothelial cells: identification of nuclear factors that recognise functional elements in the tissue-type plasminogen-activator gene promoter. AB - The gene encoding human tissue-type plasminogen activator (t-PA) is regulated in a cell-type-specific manner. Previous studies in non-endothelial cells have indicated that basal and phorbol ester mediated induction is controlled by a cAMP response element (CRE) referred to as the tPACRE, and an activating protein 2 (AP 2)-like site. The classification of the AP-2-like site was assigned on the basis of its sequence homology, but has been shown in some cell systems to be recognised by promoter-specific transcription factor-1 (Sp-1). Here, we have investigated the transcriptional regulation of the t-PA gene in endothelial cells and addressed the functional roles of the tPACRE and the Sp-1/AP-2-like sites. 5' RACE experiments indicate that the t-PA gene uses two transcription initiation sites in these cells with the downstream site being preferred. Functional analyses of the t-PA promoter using reporter-gene constructs transfected into C11STH endothelial cells demonstrate that the first 410 bp of the t-PA promoter confers an increase in reporter-gene activity on treatment with 4beta-phorbol 12 myristate 13-acetate (PMA). Mutagenesis of either the tPACRE or the Sp-1/AP-2 site weakens both basal and inducible expression, while disruption of both sites renders the promoter completely unresponsive. Using supershift assays, we identify the predominant tPACRE-binding proteins in nuclear extracts prepared from both C11STH cells and primary umbilical vein endothelial cells (HUVECs) as activating transcription factor 2, CREB (cAMP-responsive-element-binding protein), CREM (cAMP response element modulator) and c-jun. Treatment of cells with PMA results in a selective recruitment of jun-D to the tPACRE, while Sp-1 was identified as the major transcription factor that recognises the AP-2-like site. Based on this data and previous reports, we have reassigned this as a Sp-1 binding site. Finally, the identification of specific endothelial-derived t-PACRE binding proteins suggests an integral role for these factors in the regulation of t-PA gene expression in human endothelial cells. PMID- 9851701 TI - MtDNA mutations associated with sideroblastic anaemia cause a defect of mitochondrial cytochrome c oxidase. AB - We have recently described heteroplasmic mutations of mitochondrial DNA in patients suffering from sideroblastic anaemia. The mutations change conserved residues 1280 and M273 in subunit I of cytochrome oxidase, the terminal enzyme of the mitochondrial respiratory chain. As a step towards elucidating the pathogenic mechanism, we studied the biochemical consequences of the mutations by transferring mtDNA from these patients' platelets into a permanent human cell line lacking a mitochondrial genome. Mutation-induced changes of the enzyme and the energy metabolism of the cells were characterised in the transmitochondrial cell lines. One of the mutations resulted in a decreased cellular concentration of the enzyme and a corresponding decrease in activity. The second mutation changed the structure around the binuclear centre and forced the cells to rely more strongly on glycolysis. PMID- 9851702 TI - Structural determination of the O-antigenic polysaccharide from Escherichia coli O35 and cross-reactivity to Salmonella arizonae O62. AB - The structure of the O-antigenic polysaccharide from Escherichia coli O35 has been investigated with the aid of NMR spectroscopy, sugar and methylation analyses. The sequence of the sugar residues could be determined by NOESY and heteronuclear-multiple-bond-connectivity NMR experiments. The polysaccharide is composed of hexasaccharide repeating units with the following structure, where Rha and GalNAcAN represent rhamnose and 2-acetamido-2-deoxy-galacturonamide, respectively: carbohydrate sequence [see text]. The O-antigen of Escherichia coli O35 is similar to the O-specific polysaccharide from Salmonella arizonae O62, which instead has a terminal 2-acetamido-2-deoxy-alpha-D-galacturonic acid residue. Immunochemical analyses using a rabbit antiserum specific for the Salmonella arizonae O62 O-antigen showed an identical reactivity with both lipopolysaccharides. PMID- 9851703 TI - Stimulation of Ca2+-dependent exocytosis and arachidonic acid release in cultured mast cells (RBL-2H3) by a GTPase-deficient mutant of G alpha i3. AB - Gi3, a member of the Gi family of heterotrimeric GTP-binding proteins, regulates vesicle trafficking along both the constitutive and regulated pathways. In mast cells, specialized secretory cells which secrete a variety of inflammatory mediators by regulated exocytosis, activation of Gi3 provides a sufficient signal for exocytosis [Aridor, M., Rajmilevich, G., Beaven, M. A. & Sagi-Eisenberg, R. (1993) Science 262, 1569-1572]. Such activation can be achieved in patch-clamped or streptolysin-O (SLO)-permeabilized mast cells by a combination of Ca2+ and nonhydrolyzable analogs of GTP. In contrast, Ca2+-activated exocytosis in intact cells is Gi3 independent. We show here that overexpression of a GTPase-deficient mutant (G alpha i3Q204L), but not of the wild-type form of G alpha i3, in rat basophilic leukemia cells (RBL-2H3), a tumor analog of mucosal mast cells, resulted in marked potentiation of exocytosis and release of arachidonic acid in intact cells activated by a Ca2+ ionophore alone or in combination with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate. In contrast, exocytosis and arachidonic acid release stimulated by aggregation of the cell surface receptors for immunoglobulin E (IgE) were unaffected. These results strongly suggest that the intracellular receptor, responsible for the activation of Gi3, is a low affinity Ca2+-binding protein that can only be activated during Ca2+ ionophore stimulation. Moreover, these results also suggest that the propagation of the Ca2+-activated and Gi3-mediated signaling pathway requires the blocking of Gi3 GTPase activity. Finally, our results indicate that release of arachidonic acid is at least one of the downstream effectors of Gi3. PMID- 9851704 TI - Lipophosphoglycan of Leishmania donovani inhibits lipid vesicle fusion induced by the N-terminal extremity of viral fusogenic simian immunodeficiency virus protein. AB - Lipophosphoglycan (LPG), the major glycoconjugate of Leishmania parasites, was recently shown to be a potent inhibitor of viral infection. The mechanism by which this natural membrane amphiphile compound inhibits membrane fusion was investigated in this study using a simple model membrane system and a synthetic peptide corresponding to the fusion peptide of simian immunodeficiency virus (SIV). At low concentration (< 10 microM), LPG inhibits SIV-induced lipid mixing of large unilamellar vesicles composed of an equimolar mixture of egg phosphatidylcholine and egg phosphatidylethanolamine. Importantly, this inhibition was observed regardless of which LPG was inserted in the inner monolayer, the outer monolayer or both sides of the membrane, suggesting that the inner monolayer plays a determining role in membrane fusion. Fourier transform infrared spectroscopy revealed that LPG induced a conformational change of SIV fusion peptide without affecting its capacity to interact with the lipid membrane. This structural change was shown not to depend on the LPG localization and was observed even when LPG was exclusively associated to the inner lipid membrane. PMID- 9851705 TI - The N-terminal region of non-A beta component of Alzheimer's disease amyloid is responsible for its tendency to assume beta-sheet and aggregate to form fibrils. AB - Examination of the N-terminal sequence of non-A beta component of Alzheimer's Disease amyloid (NAC) revealed a degree of similarity to regions crucial for aggregation and toxicity of three other amyloidogenic proteins, namely amyloid beta peptide (A beta), prion protein (PrP) and islet amyloid polypeptide (IAPP), leading us to believe that this might be the part of the molecule responsible for causing aggregation. Secondary structure prediction analysis of NAC indicated that the N-terminal half was likely to form a beta-structure whereas the C terminal half was likely to form an alpha-helix. NAC in solution altered from random coil to beta-sheet structure upon ageing, a process that has previously been shown to lead to fibril formation. To delineate the region of NAC responsible for aggregation we synthesised two fragments, NAC-(1-18)-peptide and NAC-(19-35)-peptide, and examined their physicochemical properties. Upon incubation, solutions of NAC-(1-18)-peptide became congophilic and aggregated to form fibrils of diameter 5-10 nm, whereas NAC-(19-35)-peptide did not bind Congo Red and remained in solution. Circular dichroism spectroscopy was used to study the secondary structure of NAC and the two fragments. In trifluoroethanol/water mixtures, NAC and NAC-(19-35)-peptide adopted alpha-helical structure but NAC-(1 18)-peptide did not. NAC-(1-18)-peptide and NAC formed beta-sheet in acetonitrile/water mixtures more readily than did NAC-(19-35)-peptide. CD spectra of NAC or NAC-(1-18)-peptide in aqueous solution indicate the formation of beta sheet on ageing. We propose that the N-terminal region of NAC is the principal determinant of aggregation. Our results indicate that NAC resembles A beta, and other amyloidogenic proteins, in that aggregation is dependent upon beta-sheet development. These results lend support to a role for NAC in the development of neurodegenerative disease. PMID- 9851706 TI - Rapid analysis of epitope-paratope interactions between HIV-1 and a 17-amino-acid neutralizing microantibody by electrospray ionization mass spectrometry. AB - Progress in therapeutic or prophylactic immune intervention in HIV-1 infections may only come about with a detailed understanding at the molecular/atomic level of how antibodies neutralize (inactivate) virus infectivity. Currently information on the molecular aspects of antibody-virus interaction comes predominantly from X-ray crystallography, a process that is dependent on the production of suitable crystals. NMR can also be valuable but is complex and time consuming, while mass spectrometry has been limited to matrix-assisted laser desorption ionization (MALDI) analysis of peptides eluted from the cognate antibody. Here, we have used electrospray ionization mass spectrometry (ESI-MS) to detect directly the interactions of a novel 17-amino-acid microantibody (MicroAb) that has HIV-1-inhibitory activity, and peptides representing the V3 regions of primary HIV-1 strains isolated from Brazil (clade B) and Africa (clade A). The MicroAb is based on the third complementarity-determining region of the heavy chain (CDR-H3) of a murine monoclonal IGGI (F58) specific for the V3 loop of the gp120 envelope glycoprotein of HIV-1. ESI-MS proved to be rapid (taking < 3 h for the entire analysis), sensitive (analytes at 2 mmol/ml), and accurate (RMM estimation to 0.01-0.1%). With it, we showed that the MicroAb forms complexes with the V3 peptides, implying that its antiviral activity is mediated by binding directly to the virus particle. In addition, through controlled protease digestion of the V3 peptides, we concluded that the CDR-H3 MicroAb bound to RKXXXIGPGR, a region similar to the epitope of the whole IgG as determined by ELISA. We believe that the approach exemplified here will be applicable generally to the identification of groups involved in receptor-ligand interactions. PMID- 9851707 TI - Structural alterations of alpha-crystallin during its chaperone action. AB - The small heat-shock protein, alpha-crystallin, has chaperone ability whereby it stabilises proteins under stress conditions. In this study, alterations in the structure of alpha-crystallin during its interaction with a variety of substrate proteins (insulin, alpha-lactalbumin, ovotransferrin and serum albumin) under stress conditions have been examined using visible absorption, 31P-NMR and 1H-NMR and fluorescence spectroscopy. The fluorescence and 31P-NMR data imply that during the chaperone action of alpha-crystallin under reducing conditions, there is a slight increase in hydrophilicity of its N-terminal region and an alteration in flexibility of its C-terminal region, but overall, alpha-crystallin does not undergo a gross structural change. The fluorescence data suggest that substrate proteins interact with alpha-crystallin in a molten globule or intermediately folded state. The same conclusion is made from 1H-NMR spectroscopic monitoring of the interaction of alpha-crystallin with substrate proteins, e.g. the insulin B chain. The stoichiometry of interaction between alpha-crystallin and the various substrate proteins reveals that steric factors are important in determining the efficiency of interaction between the two proteins, i.e. on a molar subunit basis, alpha-crystallin is a more efficient chaperone protein with smaller substrate proteins. Comparison is also made between the high-molecular-mass (HMM) complexes formed between alpha-crystallin and ovotransferrin when reduced and heat stressed. Under heating conditions, fluorescence spectroscopy indicates that the HMM complex has a greater exposure of hydrophobicity to solution than that formed by reduction. Furthermore, in interacting with heated ovotransferrin, the C-terminal extension of the alphaB-crystallin subunit preferentially loses its flexibility suggesting that it is involved in stabilising bound ovotransferrin. By contrast, this extension is only partially reduced in flexibility in the HMM complex formed after reduction of ovotransferrin. The functional role of the C terminal extensions in the chaperone action and the overall quaternary structure of alpha-crystallin is discussed. PMID- 9851708 TI - Protein engineering of the restriction endonuclease EcoRV--structure-guided design of enzyme variants that recognize the base pairs flanking the recognition site. AB - We generated variants of the restriction endonuclease EcoRV that discriminate between recognition sites with different flanking sequences. This was achieved by designing new contacts to the bases in the major groove of the DNA preceding and following the EcoRV recognition site. We selected Ala181 as the starting point for the extension of the site specificity of EcoRV because, according to the structure of the specific EcoRV x DNA complex, this residue is involved in a water mediated contact with the bases flanking the recognition sequence on the 5' side. A substitution of this alanine residue by other amino acid residues changes the protein-DNA interface in this region and potentially creates new contacts, such that EcoRV variants could have an extended specificity, i.e. a greater selectivity for EcoRV sites within a particular sequence context. EcoRV variants with naturally occurring amino acid residues at position 181 were produced and their selectivity analyzed with oligodeoxynucleotide and plasmid substrates that differ only in the base pairs immediately flanking the EcoRV site. Some variants, having amino acid residues with long or bulky side chains at position 181 showed altered preferences for the base pairs flanking the recognition sequence with oligodeoxynucleotide substrates without loosing their catalytic efficiency. One variant, A181K, is able to discriminate between purine and pyrimidine bases on the 5' side of the recognition sequence, probably by means of a new hydrogen bond to the N7 of the purine base. Another variant, A181E, strongly prefers a thymine base on the 5' side of the recognition sequence, presumably due to a hydrogen bond formed between the protonated glutamic acid residue and the O4 of thymine. PMID- 9851709 TI - Side-chain selective and covalent labelling of proteins by organometallic complexes of heavy transition metals--possible application in radio crystallography of proteins. AB - Organo-rhenium and organo-tungsten N-succinimidyl and N-sulfosuccinimidyl esters react at amino groups of proteins to form stable amide bonds between the protein and the heavy transition metal complexes. We show here that factors such as pH of the reaction medium and nature of the reagent (coordinating metal, surrounding ligands, type of ester) had a marked influence on the final number of coupled organometallic groups per protein molecule, defined as the coupling ratio. By operating with stoichiometric quantities of reagent with respect to the number of aqino groups, up to 30 organometallic groups were introduced into the model protein, bovine serum albumin (BSA). BSA conjugates were characterized by gel electrophoresis in non-denaturating conditions. Hen egg-white lysozyme organo tungsten conjugates were prepared in the same manner and the peptides resulting from their tryptic hydrolysis were analyzed by reverse-phase HPLC. A tentative assignment of the preferential amino sites of conjugation is suggested from the latter results. PMID- 9851710 TI - Progress curves--a mean for functional classification of cellulases. AB - The chain-end preference and processivity of the cellulases 1,4-beta-D-glucan cellobiohydrolase I (CBH I) and 1,4-beta-D-glucan-cellobiohydrolase II (CBH II) from Trichoderma reesei and 1,4-beta-D-glucan-cellobiohydrolase 50 (CBH 50) and 1,4-beta-D-glucan-cellobiohydrolase 58 (CBH 58) from Phanerochaete chrysosporium were studied by comparing experimental degradation data on reducing end-labelled bacterial microcrystalline cellulose with computer simulations of different models. Our results with T. reesei and P. chrysosporium cellulases show that there is a common pattern of hydrolysis for CBH-I-type enzymes and another clearly distinguishable pattern for CBH-II-type enzymes, especially in the initial part of the progress curve. PMID- 9851711 TI - Site-directed mutagenesis studies of the metal-binding center of the iron dependent propanediol oxidoreductase from Escherichia coli. AB - The amino acid residues involved in the metal-binding site in the iron-containing dehydrogenase family were characterized by the site-directed mutagenesis of selected candidate residues of propanediol oxidoreductase from Escherichia coli. Based on the findings that mutations H263R, H267A and H277A resulted in iron deficient propanediol oxidoreductases without catalytic activity, we identified three conserved His residues as iron ligands, which also bind zinc. The Cys362, a residue highly conserved among these dehydrogenases, was considered another possible ligand by comparison with the sequences of the medium-chain dehydrogenases. Mutation of Cys362 to Ile, resulted in an active enzyme that was still able to bind iron, with minor changes in the Km values and decreased thermal stability. Furthermore, in an attempt to produce an enzyme specific only for the zinc ion, three mutations were designed to mimic the catalytic zinc binding site of the medium-chain dehydrogenases: (1) V262C produced an enzyme with altered kinetic parameters which nevertheless retained a significant ability to bind both metals, (2) the double mutant V262C-M265D was inactive and too unstable to allow purification, and (3) the insertion of a cysteine at position 263 resulted in a catalytically inactive enzyme without iron-binding capacity, while retaining the ability to bind zinc. This mutation could represent a conceivable model of one of the steps in the evolution from iron to zinc dependent dehydrogenases. PMID- 9851712 TI - Papaya glutamine cyclase, a plant enzyme highly resistant to proteolysis, adopts an all-beta conformation. AB - Glutamine cyclases catalyse the conversion of L-glutaminyl-peptides into 5 oxoprolyl-peptides with the concomitant liberation of ammonia. We report here biophysical characterisation of the glutamine cyclase present in the laticiferous cells of the plant Carica papaya. After purification to near homogeneity, this enzyme was subjected to limited proteolysis and found to exhibit a high resistance to degradation and nicking. The structural reasons for this property were examined using circular dichroism and infrared spectroscopies. By combining the analyses of the infrared and CD spectra of papaya glutamine cyclase, its susceptibility to proteolysis, and its hydrogen-deuterium exchange characteristics, we conclude that this protein contains extensive beta-sheet structure and is likely to have only short immobile loops connecting its beta strands. PMID- 9851713 TI - Cloning and biochemical characterisation of Aspergillus niger hexokinase--the enzyme is strongly inhibited by physiological concentrations of trehalose 6 phosphate. AB - The Aspergillus niger hexokinase gene hxkA has been cloned by heterologous hybridisation using the Aspergillus nidulans hexokinase gene as a probe. The DNA sequence of the gene was determined, and the deduced amino acid sequence showed significant similarity to other eukaryotic hexokinase and glucokinase proteins, in particular to those of the budding yeasts. The encoded protein was purified from a multicopy hxkA transformant, and extensively characterised. The hexokinase protein has a molecular mass of 54090, a pI of 4.9 and is a homodimer. D-Glucose, the glucose analogue 2-deoxy-D-glucose, D-fructose, D-mannose and D-glucosamine are phosphorylated by hexokinase, whereas the hexoses D-galactose, L-sorbose, methyl alpha-D-glucoside and the pentoses L-arabinose and D-xylose are not. The enzyme has high affinity for glucose (Km = 0.35 mM at pH 7.5) and for fructose (Km = 2.0 mM at pH 7.5) and is inhibited by ADP. The enzyme is strongly inhibited by physiological concentrations (0.1-0.2 mM) of trehalose 6-phosphate, which may be of importance for in vivo regulation of the enzyme. Inhibition of A. niger hexokinase by trehalose 6-phosphate is competitive towards the sugar substrate (Ki = 0.01 mM). Based on the kinetic constants of hexokinase and glucokinase their relative contribution to in vivo glucose phosphorylation was calculated and found to be strongly dependent on intracellular pH and glucose concentration. At pH 7.5 glucokinase is predominant, whereas at pH 6.5 hexokinase is predominant at glucose concentrations higher than 0.5 mM. Expression of the hexokinase and the glucokinase gene requires active carbon metabolism. Also on carbon sources which are not substrates for hexokinase or glucokinase, clear expression is observed. The hexokinase and glucokinase enzymes are quite stable in vivo. Even in the absence of transcription, active glucokinase and hexokinase remain present in the cells at almost the same level for at least 3-4 h after depletion of the carbon source. PMID- 9851714 TI - Modulation of carcinoembryonic antigen release by glucosylceramide--implications for HT29 cell differentiation. AB - Previous work suggested that glucosylceramide (GlcCer) plays a role in the regulation of cell differentiation of HT29 human colon tumor cells. In the present study, we investigated the role of GlcCer in the cellular release of carcinoembryonic antigen (CEA), a marker for cell differentiation. This was done by modulating the intracellular level of the glycolipid, according to two different approaches. The cells were treated with D,L-threo-1-phenyl-2 decanoylamino-3-morpholino-1-propanol (PDMP), which resulted in a specific lowering of the cellular GlcCer pool. Alternatively, by exogenous addition of a short-chain analog of the lipid, hexanoyl(C6)-GlcCer, the cellular pool was enhanced. The results demonstrate that PDMP causes an increase in the release of CEA, while exogenous C6-GlcCer suppresses its release. Furthermore, the enhanced release of CEA in the presence of PDMP, could be completely reversed upon exogenous addition of C6-GlcCer. Control experiments reveal that a potential interference of the well-known modulator of cell physiology, ceramide (Cer), can be excluded. Long-term depletion of GlcCer resulted in a change in a morphological feature of differentiation of the cells, i.e. an increase in apical membrane surface with microvilli brush borders, accompanied by an enhanced expression of the cytoskeletal protein villin. These results, together with the observations on modulation of the differentiation marker CEA by GlcCer, provide support for the conclusion that GlcCer interferes with the differentiation of HT29 cells. PMID- 9851716 TI - Fibroblast growth factor-2 has opposite effects on human breast cancer MCF-7 cell growth depending on the activation level of the mitogen-activated protein kinase pathway. AB - In human breast cancer MCF-7 and MCF-7ras cells, we demonstrated that whereas insulin had a mitogenic effect on both cell lines, fibroblast growth factor-2 (FGF-2) had opposite effects, stimulating MCF-7 and inhibiting MCF-7ras cell proliferation. The inhibitory signal induced by FGF-2 was related to sustained mitogen-activated protein kinase (MAPK) activation in MCF-7ras cells, while transient MAPK activation was associated with MCF-7 cell proliferation. FGF-2 was further used in combination with insulin or cAMP. In MCF-7 cells, insulin and cAMP reversed the mitogenic effect of FGF-2. In MCF-7ras cells, insulin did not modify the inhibitory effect of FGF-2, but cAMP markedly enhanced it. These effects were also associated with an increased level and duration of MAPK activation. PD98056 abolished the effect of FGF-2 on DNA synthesis in both cell lines, demonstrating that the dual effect of FGF-2 on cell proliferation is dependent on the activity of the extracellular-signal-regulated kinase 1 and 2 (ERK1/2) signalling pathway. PMID- 9851715 TI - Sialylated N-glycans in adult rat brain tissue--a widespread distribution of disialylated antennae in complex and hybrid structures. AB - This paper extends our earlier work on the analysis of neutral N-glycans from adult rat brain to glycans carrying NeuAc residues as their sole charged groups. These structures comprised at least 40% of the total (acidic and neutral) N glycan pool. Compounds were identified by a combination of endoglycosidase and exoglycosidase digestions, anion-exchange chromatography, normal and reverse phase high-performance liquid chromatography, matrix-assisted laser desorption/ionisation-mass spectrometry and combined gas chromatography/mass spectrometry. Mono-, di- and trisialylated components, together with components substituted with four (or more) NeuAc residues, showed abundances of approximately 12, 10, 7 and 7%, respectively, relative to the total N-glycan pool. In addition, neuraminidase digestion resulted in the neutralisation of a fraction of highly charged species, possibly indicating the presence of N-glycans substituted with short chains of polysialic acid. Sialylated bi-, tri- [mainly the (2,4)-branched isomer], tetraantennary complex, polylactosamine and hybrid structures were detected. Typically, for 'brain-type' N-glycosylation, these sialylated structures were variously modified by the presence of core alpha1-6 linked and outer-arm alpha1-3-linked fucose residues and by a bisecting GlcNAc. Structural groups such as sialyl Lewis(x) and NeuAc alpha2-3 substituted Galbeta1 4GlcNAc antennae were common. In contrast to the neutral glycans, however, a widespread distribution of terminal beta1-3-linked galactose residues was observed. The presence of beta1-3-linked galactose allowed for a high degree of sialylation as afforded by the presence of the NeuAc alpha2-3Galbeta1-3(NeuAc alpha2-6)GlcNAc structural group. This revealed a number of novel structures including the presence of tetraantennary N-glycans with more than one beta1 3galactose residue and (2,4)-branched triantennary oligosaccharides containing three such residues. Disialylated hybrid glycans containing beta1-3-linked galactose and 'polylactosamine' N-glycans with one to three terminal beta1 3galactose residues were additional novel features. The N-glycans modified by polysialylation lacked outer-arm fucose and bisecting GlcNAc residues but all contained one or more terminal beta1-3-linked galactose residues. These may be representative, therefore, of the polysialylated N-glycans expressed mainly on neural cell-adhesion molecules and known to be present in adult rat brain. The diversity of presentation of terminal sialylated groups in rat brain implies potential specificity for possible charge or lectin-mediated interactions. The distinguishing sets of sialylated structures described here are indicative of differences in the natural glycosylation processing pathways in different cell types within the central nervous system, a specificity that may be further magnified on the individual glycoproteins. PMID- 9851717 TI - Characterization of soybean seed coat peroxidase: resonance Raman evidence for a structure-based classification of plant peroxidases. AB - Electronic absorption and resonance Raman spectra of ferric and ferrous forms of a peroxidase from soybean seed coat (SBP) at neutral and alkaline pH values together with the spectra of the ferric-fluoride complex are reported. At neutral pH a quantum mechanically mixed spin state, resulting from the admixture of intermediate spin, S = 3/2, and high spin, S = 5/2, configurations, has been identified which coexists with five- and six-coordinate high-spin hemes. A complete conversion to a fluoride-ligated six-coordinate high-spin and a hydroxy ligated six-coordinate low-spin heme are observed at acid pH in the presence of fluoride and at alkaline pH, respectively. The spectral features suggest that both the fluoride and hydroxo ligands are stabilized by hydrogen-bond interactions with the distal Arg residue and through a water molecule with the distal His residue. The ferrous form shows a single nu(Fe-Im) at 246 cm(-1) at neutral pH. The data indicate that SBP shares many characteristics with peroxidases belonging to class III of the "plant peroxidase" superfamily. PMID- 9851718 TI - Resonance Raman spectroscopic study of the caa3 oxidase from Thermus thermophilus. AB - The terminal caa3 oxidase of Thermus thermophilus has been studied by resonance Raman spectroscopy. Using different excitation wavelengths in the Soret band region, it was possible to disentangle the resonance Raman spectra of the fully oxidized and fully reduced state in terms of the component spectra of the individual hemes a, a3, and c. For the heme a and a3 groups, the spectra reveal only minor differences compared to those of beef heart cytochrome c oxidase attributable to subtle modifications of the heme environment. These differences are not more pronounced than those between the oxidases from beef heart and Paracoccus denitrificans confirming the view that this oxidase of Th. thermophilus is a typical member of the aa3 oxidase superfamily. The heme c component spectra display far-reaching similarities with those of c-type cytochromes which serve as mobile electron carriers in the respiratory chain. These results imply that caa3 oxidase represents an integrated version of the noncovalent redox complex between cytochrome c and cytochrome c oxidase in higher organisms. On the other hand, the structural changes of cytochrome c in the noncovalent complex have no counterpart in the heme c component of the caa3 oxidase indicating a specific cytochrome c binding site for the mitochondrial enzyme. PMID- 9851719 TI - On the use of ultraviolet resonance Raman intensities to elaborate molecular force fields: application to nucleic acid bases and aromatic amino acid residues models. AB - Normal modes analyses for different molecules with biological interest have been performed and checked via the calculation of resonance Raman intensities. For this purpose, molecular orbital calculations were used to determine bond order changes in the lowest-lying electronic transitions. These bond order changes were used to calculate resonance Raman intensities in order to obtain correct vibrational assignments and reliable force fields. PMID- 9851721 TI - Composition, constitution, and interaction of bone with hydroxyapatite coatings determined by FT Raman microscopy. AB - An optimized FT Raman microscope (inverted microscope with high throughput of radiation) was developed that allows minimal sample preparation and Raman spectroscopy without fluorescence. A quantitative determination of the mineralization of bone tissue and hydroxyapatite (HA) coatings of hip and knee prostheses was performed. The lateral resolution reached down to 10 microm. The distribution of the HA content in the coatings investigated was found to be similar all the time. This result was independent of the composition of the coatings and the history of the whole prosthesis. In the immediate vicinity of the prosthesis a large HA content could be observed that decreased to a minimum towards the periphery of the coating and increased at the site of the ongrown bone. For the interface between bone and HA coating a transitional zone was observed at a lateral distance of 30-40 microm to the implant. PMID- 9851720 TI - Fluorescent properties of amino acids labeled with ortho-aminobenzoic acid. AB - ortho-Aminobenzoic acid (Abz) has been used as a convenient fluorescent donor group in internally quenched fluorescent peptides, which are employed as substrates for several proteolytic enzymes. As Abz is usually bound to the N amino terminal of these peptides, it is of interest to investigate the Abz group fluorescent properties bound to different amino acids. We report in this article the optical absorption and fluorescent properties, in aqueous media, of Abz bound to the alpha-amino group of Ala, Gly, Leu, Ile, Val, Pro, Phe, Arg, Glu, Met, Asn, Tyr, and Trp, with monomethylamidated alpha-carboxyl group. In order to explore the origin of the drastic reduction of Abz attached to Nalpha amino group of prolyl-peptides, we also examined the fluorescence properties of Abz-NHCH3, Abz-N(CH3)2, and Abz-pyrrolidine. Molecular dynamics simulation and NMR data indicated a lack of periplanarity of the Abz-dimethylamide, which could be the origin of low fluorescence quantum yield of Abz-prolyl-peptides. PMID- 9851722 TI - Applications of Raman spectroscopy to ophthalmology: spectroscopic characterization of disposable soft contact lenses. AB - Disposable soft contact lenses based on HEMA-MAA hydrogels are examined using Raman and ATR/FTIR vibrational spectroscopies and thermal analysis. The main factors dealing with physical, chemical, and biological biocompatibility are evaluated in relation to those of long-wear soft contact lenses with the aim of individuating the most biocompatible lens. The Raman spectra of HEMA-MAA lenses show that some biocompatibility factors are affected by environmental conditions and, in particular, by changes in pH and ionic strength values. PMID- 9851723 TI - Mechanism of action of intravenous immunoglobulin and therapeutic considerations in the treatment of autoimmune neurologic diseases. AB - On the basis of controlled clinical trials, high-dose intravenous immunoglobulin (IVIg) has emerged as a critical therapy in the management of patients with various autoimmune neurologic diseases. Different interpretations of the clinical trial results, the expected benefit of IVIg compared to that of alternate therapies, off-label use, results from anecdotal or empirical clinical trials, issues about safety and cost and, most importantly, the uncertainty about mechanisms of action of IVIg have generated concerns among practicing neurologists. Understanding the mechanisms of action of IVIg and its potential risks or side effects is expected to help us make more judicious use of this powerful immunodulating agent. This article provides evidence that IVIg has multiple actions which may operate in concert with each other but that for each disease a predominant mechanism may prevail according to the underlying immunopathogenetic cause of the respective disorder. The most relevant actions of IVIg in the therapy of neurological diseases include (a) inhibition of complement binding and prevention of MAC formation, (b) neutralization of certain pathogenic cytokines, (c) downregulation of antibody production, and (d) modulation of Fc receptor mediated phagocytosis. Additional actions include the effect of Mg on superantigens, modulation of T-cell function and antigen recognition, and enhancement of remyelination. On the basis of our experience with more than 130 closely monitored patients, I provide guidelines on how to use the drug, kinetics of IVIg, adverse reactions, and risk factors. In addition, the incidence, morbidity prevention and treatment of the common or rare side effects, including thromboembolic events, increased serum viscosity, aseptic meningitis, headaches, skin reactions, renal failure, and hemolysis are described. Spurious results of serologic tests, such as false hyponatremia and elevated sedimentation rate, as well as a transient rise in various viral titers, are described in an effort to avoid misinterpretations by treating neurologists. Finally, details on the latest findings of viral safety of IVIg are provided. PMID- 9851724 TI - Treatment of Guillain-Barre syndrome with intravenous immunoglobulin. AB - Guillain-Barre syndrome (GBS) is an acute polyneuropathy that typically presents as a progressive flaccid paralysis. The pathology is believed to be caused by both cellular and humoral immune processes. This inflammatory neuropathy has a mortality rate of 4-5%. About 30% of patients require mechanical ventilation, and these patients are often hospitalized for months before regaining the ability to walk. Immunomodulation is used to improve the recovery rate and shorten hospital stays. Plasma exchange was shown to be effective in improving recovery time in GBS in several controlled trials during the 1980s. In this decade, intravenous immunoglobulin (IVIg) therapy has been shown to be equally effective for therapy of GBS and its variants. Although the precise mechanisms of immunomodulation by IVIg are unknown, it probably directly inactivates specific anti-myelin antibodies and indirectly inhibits their production. IVIg offers some advantages over plasma exchange by being better tolerated in some patients and being easily administered without special equipment. However, because of the possibility of progression, the treatment of GBS patients requires qualified neurologic and supportive care. PMID- 9851725 TI - Treatment of chronic inflammatory demyelinating polyneuropathy with intravenous immunoglobulin. AB - Chronic inflammatory demyelinating polyneuropathy (CIDP) is a defined clinical entity with a chronic progressive or chronic relapsing course, lasting months to years. It causes variable but often severe chronic disability. CIDP is considered an autoimmune disorder caused by both cellular and humoral immune processes. Various immunomodulatory therapies, i.e., IVIg, therapeutic plasma exchange (PE), and prednisone, are of proven benefit. Comparative studies indicate that IVIg and PE confer equal short-term benefit. Efficacy of IVIg is maintained; regularly timed pulse treatments may stabilize relapsing CIDP. The combination of IVIg and prednisone may be advantageous in long-term management. Despite the high cost, IVIg is considered the preferred first treatment. The safety profile is similar to that reported for other conditions; close monitoring during the infusion is recommended. The precise mechanisms of IVIg action in CIDP are not known. Anti idiotypic neutralization of autoantibodies, binding of complement, and blockade of macrophages may prevent the ongoing inflammatory demyelination. PMID- 9851726 TI - Multifocal motor neuropathy: diagnosis and treatment. AB - Multifocal motor neuropathy (MMN) and some lower motor neuron syndromes are immune-mediated and treatable. These motor syndromes produce weakness that is typically distal and asymmetric, involves the arms early in the course of disease, and progresses slowly. Electrophysiologic abnormalities often include evidence of demyelination, especially focal conduction block, selectively on motor axons. High titers of serum IgM binding to GM1 ganglioside, alone or in a membrane environment, occur in 80-90% of patients with MMN. Treatments for MMN that commonly produce increased strength include IV human immune globulin (HIG) and cyclophosphamide. After an initial treatment with 2 g/kg of HIG, up to 80% of patients with MMN show short-term improvement. Long-term HIG treatment is useful in 60% of MMN patients and has few side effects but is costly. Intravenous cyclophosphamide treatment is effective in 70% of MMN patients but has significant toxicity, and is reserved for patients who have severe disease and do not respond adequately to HIG. PMID- 9851727 TI - Intravenous immunoglobulins in multiple sclerosis. AB - The spectrum of diseases being treated with intravenous immunoglobulins (IVIg) appears to be ever broadening, including use in neurologic diseases. After a period of anectodal reports and smaller uncontrolled series, recently there have been several randomized, prospective, double-blind, placebo-controlled studies employing IVIg in patients with relapsing remitting (RR) multiple sclerosis (MS). Reduction in relapse rate and some evidence of decreased MRI activity has been reported, but to date no effect on disability or MRI lesion burden has been noted. Because of differences in methodologic design and patient populations, as well as the relatively small number of patients in some of these studies, a rigorous direct comparison of efficacy with the type I interferons and glatiramer acetate is not possible. Given these data and the high cost of IVIg, routine use of this mode of therapy cannot be recommended, certainly not as a first-line treatment. Larger studies would clearly be helpful. At present there is no evidence to support the use of IVIg in secondary progressive (SP) or primary progressive (PP) MS. PMID- 9851728 TI - Intravenous immunoglobulin for the treatment of acquired myasthenia gravis. AB - Acquired myasthenia gravis (MG) is an autoimmune disorder characterized by exertional fatigue and weakness that is made worse with activity and improved with rest, only to recur with the resumption of activity. The pathology results from an antibody-mediated attack to several different epitopes of the acetylcholine receptor (AChR) complex. The consensus of an expert panel is that intravenous immunoglobulin (IVIg) is effective in reversing myasthenic weakness. Although the mechanism of action is not known, it is likely that there is a downregulation of antibody production. IVIg appears to have a role as an acute treatment intervention in rapidly progressive weakness or as a chronic maintenance therapy when all other treatment modalities have failed. Its response is similar to but slower than the response of plasma exchange (PE), but it offers advantages when therapeutic apheresis is not available or when vascular access is problematic. PMID- 9851729 TI - Controlled studies with high-dose intravenous immunoglobulin in the treatment of dermatomyositis, inclusion body myositis, and polymyositis. AB - There are three major subsets of the inflammatory myopathies: polymyositis (PM), dermatomyositis (DM), and inclusion-body myositis (IBM). High-dose intravenous immunoglobulin (IVIg) has been tried in controlled clinical trials in patients with DM and IBM but not with PM. In patients with DM that is resistant or partially responsive to conventional therapies, IVIg was very effective. The treated patients experienced dramatic improvement not only in muscle strength but also of their skin rash. Repeated muscle biopsies with quantitative histologic studies showed the IVIg-treated patients had a statistically significant improvement of the muscle cytoarchitecture, with resolution of the aberrant immunopathologic parameters. In two controlled clinical trials conducted in IBM patients, IVIg showed marginal improvements in muscle strength which were nonsignificant. However, a few IBM patients had a definite clinical improvement with increased activities of daily living, but when analyzed within the entire IVIg-treated group, their total gains in muscle strength did not reach statistical significance compared to the placebo-treated group. Of interest is that certain muscle groups in the IVIg-treated patients, such as the muscles of swallowing, showed significant improvement compared to those of the placebo treated patients, implying mild regional effects. In PM, uncontrolled trials have shown improvements in muscle strength, but the controlled clinical trial is still ongoing. PMID- 9851730 TI - Novel inherited risk factors for venous thrombosis. PMID- 9851731 TI - Aspirin analogs as antithrombotic agents. PMID- 9851732 TI - Soluble CD27 in thyroid disorders. PMID- 9851733 TI - Prevalence of the prothrombin 20210 G-to-A variant in blacks: infants, patients with venous thrombosis, patients with myocardial infarction, and control subjects. AB - A genetic variation in the prothrombin gene is located in the 3-untranslated region at position 20210 where a G-->A transition occurs. The prevalence of the mutation is 1% to 2% in white populations, and the mutation is associated with an increased risk of venous thrombosis and myocardial infarction. We report the prevalence of the A allele in blacks at birth; in black control subjects with no history of heart attack, stroke, or blood clots; in black patients with venous thrombosis; and in black patients with myocardial infarction. Among 318 infants, the prevalence of the A allele was 0.2% (1 heterozygote), with an exact, one sided upper 95% confidence limit of 0.7%. Among 185 control subjects the variant was absent, yielding an exact, one-sided upper 95% confidence limit of 0.8% for the A allele. The heterozygous genotype was found in 2 of 91 subjects with deep vein thrombosis and in none of 123 subjects with myocardial infarction. The very low prevalence of the A allele indicates that the prothrombin variant is not a major cause of venous thrombosis or myocardial infarction in blacks. PMID- 9851734 TI - Endothelial cell hypoxic stress proteins. AB - The vascular endothelium is an important mediator of vascular tone, inflammatory immune reactions, vascular permeability, angiogenesis, and hemostasis. Endothelial functions may be altered by changes in the local cellular environment, particularly changes in oxygen tension. The mechanisms by which endothelial cells (ECs) respond and adapt to hypoxia are unknown; however, the EC is one of the more hypoxia-tolerant mammalian cell types. Cultured ECs exposed to hypoxia up-regulate a set of stress proteins, termed hypoxia-associated proteins (HAPs), that are distinct from the classically described stress proteins induced by heat shock (heat-shock proteins, HSPs) or glucose deprivation (glucose regulated proteins, GRPs). Two of these proteins have been identified as the glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and non neuronal enolase (NNE). GAPDH expression during hypoxia is regulated primarily at the level of transcription, while the mechanism of NNE mRNA accumulation remains unclear. GAPDH, NNE, and the other HAPs are up-regulated by transitional metals and deferoxamine; however, unlike the situation with other hypoxia-regulated proteins such as erythropoietin, the up-regulation of GAPDH, NNE, and the other HAPs by hypoxia is not inhibited by carbon monoxide. Subcellular fractionation of hypoxic EC has shown that GAPDH and NNE are up-regulated in the cytoplasmic fraction as would be expected for a glycolytic enzyme; however, a protein corresponding to GAPDH is also up-regulated in the nuclear fraction. This suggests that GAPDH and perhaps NNE have functions aside from their catalytic function in glycolysis. It is unknown whether the up-regulation of GAPDH, NNE, and the other HAPs in ECs is related to the relative ability of ECs to adapt to hypoxia; however, other more-hypoxia-sensitive cells do not up-regulate HAPs. PMID- 9851735 TI - Mammalian iron transport: an unexpected link between metal homeostasis and host defense. AB - Iron deficiency and iron overload disorders are common in clinical practice. Both can result from perturbations in the flux of iron across the absorptive intestinal enterocyte. Until recently iron transport has been poorly understood. In 1997 two independent cloning strategies identified Nramp2 (DCT1) as the first mammalian transmembrane iron transporter. In this review we discuss evidence that Nramp-related proteins play essential roles in metal homeostasis and host defense. PMID- 9851736 TI - Focused antithrombotic therapy: novel anti-platelet salicylates with reduced ulcerogenic potential and higher first-pass detoxification than aspirin in rats. AB - The use of aspirin as an anti-platelet drug is limited by its propensity to induce gastric injury and by its adverse effect on vascular prostacyclin formation. Two phenolic non-steroidal anti-inflammatory drugs (salicylic acid and diflunisal) were modified by esterification with a series of O-acyl moieties. The short-term ulcerogenic in vitro and in vivo anti-platelet properties, pharmacodynamic profiles, and extent of hepatic extraction of these phenolic esters were compared with aspirin (acetylsalicylic acid). The more lipophilic esters (longer carbon chain length in O-acyl group) show significantly less gastrotoxicity in stressed rats than does aspirin after a single oral dose. The in vitro and in vivo anti-platelet studies show that these phenolic esters inhibited (1) arachidonate-triggered human platelet aggregation and (2) thrombin stimulated rat serum thromboxane A2 production by platelets in the clotting process almost as effectively as aspirin. The hepatic extractions of these O-acyl derivatives are significantly higher than those of aspirin. The pharmacodynamic studies show that these O-acyl derivatives of salicylic acid and diflunisal probably bind to, or combine with, the same site on the platelet cyclooxygenase as aspirin. Replacing the O-acetyl group with longer chain O-acyl moiety in this series of phenolic esters markedly reduced the potential of these agents to induce short-term gastric injury but did not lessen their activity as inhibitors of platelet aggregation. These non-acetyl salicylates may therefore represent a novel class of anti-platelet drugs with less ulcerogenic potential. PMID- 9851737 TI - Soluble CD27 in thyroid disorders. AB - We measured the soluble cytokine CD27 in a variety of thyroid disorders. Soluble CD27 was increased in untreated Graves' hyperthyroidism and in euthyroid ophthalmopathy. Levels of sCD27 were normal after the establishment of euthyroidism with propylthiouracil (PTU) or radio iodine in primary hypothyroidism, chronic thyroiditis, and the hyperthyroid and euthyroid phases of subacute thyroiditis. Soluble CD27 is a marker for cellular activation as in Graves' hyperthyroidism, but it is not predictive of the outcome of PTU therapy. PMID- 9851738 TI - Relationship of body water compartments to age, race, and fat-free mass. AB - Water compartments were studied in 72 black and 128 white women, aged 20 to 70 years. Total body water (TBW) was measured by tritiated water dilution, and extracellular water (ECW) was measured by using delayed gamma neutron activation for the determination of total body chloride. Intracellular water (ICW) was assessed as the difference between TBW and ECW. Fat-free mass (FFM) was estimated by the measurement of total body potassium (TBK) and total body fat (TBF) by measurement of total body carbon (TBC). Total body protein was calculated from total body nitrogen (TBN). TBW was found to decline with age and to also be significantly influenced by body size. The extracellular water space was 41.5% of TBW. The decline of TBW with age is due primarily to a decline in ICW. The hydration of the FFM increased with age for the overall population because of a decline in TBK and TBN and an increase in ECW. Body composition models that have assumed constancy of hydration should be adjusted not only for gender but for age and body size. PMID- 9851739 TI - Involution of keloid implants in athymic mice treated with pirfenidone or with triamcinolone. AB - The experimental drug pirfenidone (PFD) has been evaluated as an inhibitor of keloid proliferation and compared with triamcinolone (TAC) injections by studying the involution of active human keloid implants in athymic nude mice (nu-nu). PFD was fed to mice with keloid implants at a level of 2.75 mg/g of feed. At this level PFD had no adverse effect on the body weights of the mice. Implant weights in both PFD-fed and control mice decreased with time. The weights of the implants from the PFD group were significantly lower than those of the control implants at 60 and 90 days after implantation. Consequently PFD may cause an increased degradation and absorption of keloid tissue. The implants from the PFD mice were not significantly different histologically from the implants of the mice with corresponding implants. The chondroitin-4-sulfate (C4S) levels of the implants from PFD-fed mice were not significantly different from those of the implants from control mice. Therefore the mechanism of action of PFD apparently is not mediated by an effect on C4S metabolism. In contrast, the injections of TAC at a level that caused temporary body weight loss in the mice resulted in significant decreases in both hyaluronic acid (HA) and C4S in the keloid implants. Histologically, fibroblasts disappeared from the implants treated with TAC by 20 days after injection. At 30 days after TAC injection, HA and C4S were not detected by electrophoresis in keloid implants; only dermatan sulfate appeared to be present. PMID- 9851741 TI - Changes in plasma carotenoid and vitamin E profile during supplementation with oil palm fruit carotenoids. AB - It has been hypothesized that supplementation with a single source of carotenoids causes perturbations in the plasma level of diet-derived carotenoids and that this may explain the lack of association between disease rates and the intake of carotenoid supplements. This article describes the effect of supplementation with an oil palm fruit extract, rich in beta-and alpha-carotene, on the plasma carotenoid profile of 15 healthy women volunteers. Volunteers were supplemented for 35 days with 15 mg/d of total carotenoids. Blood samples were taken at regular intervals during the supplementation period and analyzed for a range of carotenoids. Results indicate that the hydrocarbon carotenoid components of the supplement are absorbed and appear in the plasma disproportionately to the ratios in the supplement and that the plasma concentration of diet-derived lutein, a dihydroxy carotenoid ((3R,3S,6R)-beta,epsilon-carotene-3,3-diol), is depressed, whereas that of lycopene is unaffected. Plasma alpha-tocopherol concentrations were unaffected by supplementation. It is concluded that supplementation with carotenoids from a single source results in plasma carotenoid profile changes that are not predictable from a knowledge of supplement composition and that such changes should be monitored and considered when drawing conclusions as to the effect of carotenoid supplementation on health outcomes. PMID- 9851740 TI - Differential regulation of human alveolar macrophage-derived interleukin-1beta and tumor necrosis factor-alpha by iron. AB - Human lungs accumulate iron with the aging process. In some circumstances associated with lung injury (eg, smoking), this acquisition of iron in lung tissue and alveolar macrophages (AMs) is escalated. We hypothesized that excess cellular iron interfered with the production of tumor necrosis factor-alpha (TNF alpha) and interleukin-1-beta (IL-1-beta) by AMs. To examine this hypothesis, we acquired AMs from smokers and nonsmokers by bronchoalveolar lavage. AMs were stimulated by lipopolysaccharide (LPS), with and without deferoxamine (DFA), a chelator of iron. Enzyme-linked immunosorbent assay and Northern analysis were used to quantitate cytokine concentrations and mRNA. The addition of DFA increased the release of IL-1-beta, but not TNF-alpha, from AMs from smokers and nonsmokers. The DFA augmentation of LPS-induced IL-1-beta was more pronounced in smokers' AMs than in those from non-smokers (4.5-fold vs 2.6-fold increase). The addition of FeCl3 to DFA diminished the augmenting effect on the release of IL-1 beta, suggesting that the mechanism of action involved iron chelation. Conversely, as the intensity of iron chelation increased, the release of IL-1 beta and TNF-alpha decreased, as was also shown with hydroxyl radical scavenging by dimethylthiourea. This inhibition, however, occurred at very different thresholds for each cytokine. These data support a relationship between excess alveolar iron and the generation of inflammation within the lung. PMID- 9851742 TI - Utility of endoscopic biopsy samples to quantitate human duodenal ion transport. AB - Duodenal mucosal bicarbonate secretion (DMBS) prevents acid-peptic damage and facilitates nutrient absorption. DMBS is diminished in patients with duodenal ulcers and is normalized after Helicobacter pylori eradication. The measurement of DMBS in human patients in vivo requires intubation with a multi-lumen balloon tube and permits limited testing with putative agonists and antagonists. Our purpose was to develop a means to investigate transport events in human duodenal biopsy samples in vitro. After validation studies in a modified mini-Ussing chamber were performed, duodenal transport events were examined in proximal endoscopic biopsy samples from normal volunteers (n = 17). Tissues were mounted in modified mini-Ussing chambers (volume 2.5 ml, surface area 3.8 mm2). Short circuit current (Isc), potential difference (PD), and bicarbonate secretion were determined under basal conditions and after stimulation with graded doses of prostaglandin E2 (PGE2)(10(-8) to 10(-4) mol/L) and dibutyryl cAMP (db-cAMP)(10( 4) to 10(-2) mol/L). Duodenal tissues remained viable for at least 2 hours and exhibited stable basal HCO3(-) secretion and electrical parameters. Stimulation with PGE2 and db-cAMP resulted in dose-related increases in both Isc and HCO3(-) secretion (P < .05) that were abolished by ouabain and anoxia. It is concluded (1) that human duodenal bulb biopsy samples maintain their inherent transport function in mini-Ussing chambers and (2) that by using this novel method it will be possible to define the transport events that modulate human duodenal secretion, in particular bicarbonate secretion, in both health and disease. PMID- 9851743 TI - Hypoxia increases thrombospondin-1 transcript and protein in cultured endothelial cells. AB - The exposure of endothelial cells to hypoxic environments regulates the expression of a number of genes with products that are vasoactive or mitogenic for vascular tissue, including platelet-derived growth factor, endothelin-1, and endothelial nitric oxide synthase. Hypoxia is also known to alter the adhesive properties of endothelium toward a variety of blood cell types. Thrombospondin-1 (TSP-1) is a glycoprotein with major roles in cellular adhesion and vascular smooth muscle proliferation and migration. We report here that hypoxia induces TSP-1 gene and protein expression. Oxygen tensions of < or =30 torr resulted in TSP-1 transcript induction initially apparent at 1 to 6 hours, with maximal induction (6.5-fold+/-1.2-fold) within 24 to 48 hours in both human and bovine endothelial cells. TSP-1 protein levels remain elevated after 72 hours of continuous hypoxic exposure. The induction of TSP-1 steady-state transcript levels is caused in large part, if not entirely, by post-transcriptional stabilization of the TSP-1 mRNA. The TSP-1 induction by hypoxia is a graded and reversible physiologic response and can be mimicked by the use of cobalt chloride or the inhibition of nitric oxide production, suggesting both the involvement of a heme-containing oxygen sensor and a role for the endogenous production of nitric oxide in TSP-1 regulation. The effects of hypoxia both on the stabilization of the TSP-1 transcript and the stimulation of TSP-1 protein production are completely inhibited by arginine butyrate. PMID- 9851744 TI - Human allogeneic lymphocytes trigger endothelial cell tissue factor expression by a tumor necrosis factor-dependent pathway. AB - Lymphocyte adhesion to endothelial cells and the extravascular deposition of fibrin are 2 important processes during pathologic situations such as allograft rejection. Tissue factor (TF) expression was therefore measured on human umbilical vein endothelial cells (HUVECs) after coculture with allogeneic lymphocytes (PBLs) by a factor Xa generation assay. When cocultured with PBLs, HUVECs expressed strong procoagulant activity related to the TF/factor VII dependent pathway, which was enhanced when endothelial cells were treated with interferon-gamma (IFN-gamma). The highest TF activity was measured when 10(5) lymphocytes were incubated with 10(4) HUVECs (ratio 10: 1) for 4 hours, a time dependent course similar to that obtained with tumor necrosis factor-alpha (TNF alpha), and direct contact between the 2 cell types was necessary. PBL-induced TF activity was inhibited by cycloheximide or actinomycin D, indicating active protein synthesis that was confirmed by the increase in TF mRNA detected by reverse transcription-polymerase chain reaction. It was then demonstrated that 1 of the primary signaling pathways leading to endothelial cell TF expression was a rapid initial interaction between membrane TNF expressed on PBLs and the 75-kd TNF receptor, with subsequent involvement of platelet-activating factor and P selectin. Finally, we showed that the transduction of external signals involving the activation of protein kinase C and protein tyrosine kinases also contributed to the regulation of TF expression. PMID- 9851745 TI - Prevalence of impaired gastric emptying of solids in systemic sclerosis: diagnostic and therapeutic implications. AB - The aims of this study were to evaluate the gastric emptying of solids in patients with progressive systemic sclerosis, correlate the esophageal motility abnormalities with their gastric emptying status, delineate the symptoms suggestive of abnormal gastric emptying, and assess the effect of metoclopramide in patients with abnormally slow gastric emptying. Twenty patients underwent esophageal motility evaluation and gastric emptying studies with a radiolabeled solid meal. Gastric emptying was also measured in 13 healthy volunteers. Four patients in whom esophageal motility was normal also had an accompanying normal rate of gastric emptying. In 16 patients with abnormal esophageal motility, mean gastric emptying was significantly delayed as compared with that in normal subjects (67.4% vs 49.8% retention of isotope at 2 hours, P < .05). Ten patients had absolute criteria for slow gastric emptying (>+2 SD). However, only postprandial bloating and early satiety were symptoms that accurately predicted slow radionuclide emptying. In four of these patients in whom gastric emptying was slow, 10 mg intramuscular metoclopramide significantly (P < .05 vs baseline) accelerated the gastric emptying of the same test meal. We conclude that (1) gastric emptying of solids was delayed in approximately two thirds of patients with abnormal esophageal motility, whereas it was normal in patients with normal esophageal motor function; (2) metoclopramide significantly accelerated this slow gastric emptying; and (3) delayed gastric emptying contributes to the severity of the gastroesophageal reflux frequently present in patients with progressive systemic sclerosis, and promotility agents offer a valuable therapeutic approach. PMID- 9851747 TI - The first commercial bottle of aspirin: March 6, 1899. PMID- 9851746 TI - Effects of glucagon on glycogenolysis and gluconeogenesis are region-specific in periportal and perivenous hepatocytes. AB - It has been established, mainly by histochemical and immunohistochemical studies, that liver cells are functionally heterogeneous, with periportal hepatocytes (PPHs) being predominantly gluconeogenic and perivenous hepatocytes (PVHs) being glycolytic. We therefore investigated the region-specific functional effects of glucagon on glycogenolysis and gluconeogenesis in isolated PPHs and PVHs prepared by the digitonin-collagenase method. BB rats, a model of insulin-dependent diabetes, were used to study the region-specific heterogeneity of gluconeogenesis in the diabetic state. Although glycogen content was not different between PVHs and PPHs in rats fed the normal diet, basal glucose release was 1.37 times greater in PVHs than in PPHs (P <.05). The increase in glucose release stimulated by 0.01 to 0.1 nmol/L glucagon was 1.52 times greater in PVHs than in PPHs (P < .05), whereas no differences were seen in response to 1 to 100 nmol/L glucagon. Glucose release from gluconeogenic substrates was 1.57 times greater in the PPHs than in the PVHs of fasted normal rats (P < .05), whereas the increase in gluconeogenesis produced by glucagon was not different between PPHs and PVHs. The glucagon-binding capacity, the cAMP release, and the increase in intracellular Ca2+ stimulated by glucagon were not different between PPHs and PVHs in the fed or fasted states. Gluconeogenesis from gluconeogenic substrates was 1.52 times greater in the PPHs than in the PVHs of fasted nondiabetic BB rats (P < .05). After the development of diabetes, the gluconeogenic capacity in PVHs increased to the level observed in PPHs, but that in PPHs did not change. Thus there was no difference in gluconeogenesis between the PPHs and PVHs of diabetic BB rats. In both the PPHs and PVHs of diabetic BB rats, the 0.01 to 100 nmol/L glucagon induced increase in gluconeogenesis was greater than that in PPHs from nondiabetic BB rats (2.30 and 3.07 times, P < .01, respectively). We conclude that PPHs and PVHs of normal rat liver express region-specific differences in their glycogenolytic and gluconeogenic responses to glucagon. In diabetic BB rats, the difference in the gluconeogenic capacity between PPHs and PVHs disappeared, whereas glucagon-induced gluconeogenesis was enhanced. PMID- 9851748 TI - Clinical review 100: Evaluation and treatment of the infertile couple. AB - Infertility by itself does not threaten physical health but has a strong impact on the psychological and social well-being of couples. In the last two decades, progress in caring for the infertile couple, in particular progress in the field of assisted reproduction and micromanipulation, has provided significant hope for many couples for whom hope could not have been offered in the past. This is especially true for bilateral tubal disease and for male factor infertility, as nearly all couples with male factor infertility can now undergo either one (or more) IVF or ICSI attempt(s). For couples with other causes of infertility, however, the differences in pregnancy rates often do not reach statistical significance. We must also remember that the total cost incurred for successful delivery for couples with a better chance of successful IVF (i.e. those with tubal disease) increases from approximately $55,000 in American dollars for the first cycle to $73,000 by the sixth cycle. Because of these high costs, many insurers in the United States and many public health systems in Europe do not cover or only partially cover these procedures. Consequently, the availability of IVF and related therapies frequently depends on the couple's ability to pay. Therefore, after having established the correct diagnosis, appropriate treatment should be counseled to the infertile couple keeping in mind the following points: 1) in subfertile couples expectant management should be reasonably counselled if the age of the woman is less than 30 yr and the duration of infertility is less than 36 months, even if oligozoospermia is present; 2) superovulation and timed intercourse seems also to be a reasonable approach in couples with anovulatory, mild/moderate endometriosis, and unexplained infertility; 3) in unexplained infertility, ovarian stimulation (with clomiphene or gonadotropin) and IUI seem to offer some advantage over ovarian stimulation and timed intercourse; 4) IVF can be a firstline approach in tubal sterility and when IUI or IPI cannot be performed because the number of motile sperm is insufficient, but is usually also the final treatment attempt when other methods have failed. The outcome of IVF is negatively influenced mainly by the woman's age; however, the number of deliveries is also generally lower in couples with male factor; 5) ICSI is a further option, which should be limited to couples: a) with very poor semen parameters; b) previous failed fertilization; c) presence of obstructive or nonobstructive azoospermia in which ICSI is combined with sperm extraction from the epididymis or the testis; 6) international register studies demonstrate that the risk of malformation after conventional IVF is not increased; 7) some reports suggest that incidence of congenital major and minor malformations is not increased in children born after ICSI. However, the rate of sex chromosome anomalies in ICSI fetuses has been reported to be approximately 1% in 585 prenatal diagnoses, a frequency increased by a factor of 4 if compared with naturally conceived live-born babies. ICSI bypasses the physiological selection of spermatozoa that occurs at the level of the testis and epididymis, and in the female reproductive tract as well as at the sperm-oocyte interface. As genetic abnormalities are present in a significant percentage of infertile males with impaired spermatogenesis, karyotyping and analysis of the Y chromosome for microdeletions should be carried out in all potential ICSI fathers. Screening for cystic fibrosis gene mutations should also be performed in azoospermia caused by congenital absence of the vas deferens and seminal vesicles. Appropriate genetic counseling should be made available to all ICSI couples whenever a gene or chromosomal anomaly has been identified. With most ARTs the average delivery rate per cycle is approximately 15% and the cumulative delivery rate after several cycles is about 50%. (ABSTRACT TRUNCATED) PMID- 9851749 TI - The diagnosis of recurrent thyroid cancer--a new approach. PMID- 9851750 TI - Germline RET codon 918 mutation in apparently isolated intestinal ganglioneuromatosis. PMID- 9851751 TI - The use of radioactive iodine in patients with papillary and follicular thyroid cancer. PMID- 9851752 TI - Three sisters with Addison's disease. PMID- 9851753 TI - Adrenocorticotropin and cortisol hyperresponsiveness to hexarelin in patients with Cushing's disease bearing a pituitary microadenoma, but not in those with macroadenoma. AB - We previously reported that in Cushing's disease (CD) the ACTH- and cortisol (F) releasing activity of Hexarelin (HEX), a GH secretagogue, is exaggerated with respect to that in normal subjects and is higher than that of human CRH (hCRH), but it is absent in Cushing's syndrome. Our aim was to extend the study about the effects of HEX (2.0 microg/kg, iv) on ACTH and F secretion in 21 patients with CD (3 men and 18 women, 16-68 yr old). Based on magnetic resonance imaging, 15 CD patients had pituitary microadenoma, and 6 had macroadenoma. The results in CD patients were compared with those in 27 normal age-matched controls (NS; 10 men and 17 women, 24-69 yr old). Basal ACTH and F levels in CD were similar in patients with microadenom (mean+/-SEM, 78.3+/-7.2 pg/mL and 237.1+/-23.6 microg/L, respectively) and macroadenoma (57.4+/-9.0 pg/mL and 196.9+/-20.1 microg/L, respectively) and were higher (P < 0.001) than those in NS (17.7+/-2.0 pg/mL and 115.3+/-6.7 microg/L, respectively). In microadenoma CD patients, HEX induced marked ACTH and F increases (delta peak, mean+/-SEM: 261.2+/-77.6 pg/mL and 226.1+/-87.2 microg/L, respectively), which were higher (P < 0.04) than those induced by hCRH (45.6+/-16.9 pg/mL and 84.6+/-25.7 microg/L, respectively). Moreover, in microadenoma CD patients, the ACTH and F responses to HEX were higher (P < 0.001) than those in NS (18.5+/-4.0 pg/mL and 36.1+/-6.8 microg/L, respectively). In macroadenoma CD patients, HEX induced a slight, but significant increase (P < 0.02) in ACTH and F levels (33.9+/-18.0 pg/mL and 89.6+/-34.3 microg/L, respectively), which was not significantly different from that elicited by hCRH (20.0+/-7.0 pg/mL and 54.8+/-21.3 microg/L, respectively). In macroadenoma CD patients, the ACTH and F responses to HEX and hCRH were, in turn, similar to those in NS. In conclusion, our findings demonstrate that the ACTH and F hyperresponsiveness to HEX is present in Cushing's disease with micro-, but not macro- ACTH-secreting pituitary adenoma. This finding agrees with other evidence pointing toward differences in the hormonal behavior between micro- and ACTH secreting pituitary macroadenomas. PMID- 9851754 TI - Prostate-sparing effects in primates of the potent androgen 7alpha-methyl-19 nortestosterone: a potential alternative to testosterone for androgen replacement and male contraception. AB - 7alpha-Methyl-19-nortestosterone (MENT) is a potent synthetic androgen that cannot be converted to dihydrotestosterone. In this study we determined the relative androgenic, antigonadotropic, and anabolic potencies of testosterone vs. MENT in the nonhuman primate M. fascicularis. In castrated monkeys, dose-response relationships were generated for the effects of testosterone and MENT on gonadotropin levels, prostate growth, body weight, and lipid metabolism. In a pilot study, four monkeys were castrated, and magnetic resonance imaging (MRI) was used to document a 50% loss of prostate volume within 8 weeks, verifying that MRI is a reliable means to measure prostate size in this species. Two additional groups of six monkeys each were then castrated and serially administered four graded dosages of testosterone or MENT via osmotic minipumps over 20 weeks. Complete suppression of LH was achieved with a minimum of 0.3 mg/day MENT, compared to 3.0 mg/day testosterone. MENT supported body weight 10 times more potently than did testosterone. Baseline prostate volumes were maintained with 0.1-0.2 mg/day MENT vs. 0.3 mg/day testosterone. Thus, in monkeys, MENT is 10 times more potent than testosterone with regard to the clinically desirable end points of gonadotropin suppression and anabolism, but only twice as potent at stimulating prostate growth. These results suggest that MENT may have a wider therapeutic index than testosterone for human androgen replacement and male contraception. PMID- 9851755 TI - High frequency of luteal phase deficiency and anovulation in recreational women runners: blunted elevation in follicle-stimulating hormone observed during luteal follicular transition. AB - The purposes of this investigation were to evaluate the characteristics of three consecutive menstrual cycles and to determine the frequency ofluteal phase deficiency (LPD) and anovulation in a sample of sedentary and moderately exercising, regularly menstruating women. For three consecutive menstrual cycles, subjects collected daily urine samples for analysis of FSH, estrone conjugates (E1C), pregnanediol-3-glucuronide (PdG), and creatinine (Cr). Sedentary (n=11) and exercising (n=24) groups were similar in age (27.0+/-1.3 yr), weight (60.3+/ 3.1 kg), gynecological age (13.8+/-1.2 yr), and menstrual cycle length (28.3+/ 0.8 days). Menstrual cycles were classified by endocrine data as ovulatory, LPD, or anovulatory. No sedentary women (0%) had inconsistent menstrual cycle classifications from cycle to cycle, but 46% of the exercising women were inconsistent. The sample prevalence of LPD in the exercising women was 48%, and the 3-month sample incidence was 79%. In the sedentary women, 90% of all menstrual cycles were ovulatory (SedOvul; n=28), whereas in the exercising women only 45% were ovulatory (ExOvul; n=30); 43% were LPD (ExLPD; n=28), and 12% were anovulatory (ExAnov; n=8). In ExLPD cycles, the follicular phase was significantly longer (17.9+/-0.7 days), and the luteal phase was significantly shorter (8.2+/-0.5 days) compared to ExOvul (14.8+/-0.9 and 12.9+/-0.3 days) and SedOvul (15.9+/-0.6 and 12.9+/-0.4 days) cycles. Luteal phase PdG excretion was lower (P < 0.001) in ExLPD (2.9+/-0.3 microg/mg Cr) and ExAnov (0.8+/-0.1 microg/mg Cr) cycles compared to SedOvul cycles (5.0+/-0.4 microg/mg Cr). ExOvul cycles also had less (P < 0.01) PdG excretion during the luteal phase (3.7+/-0.3 microg/mg Cr) than the SedOvul cycles. E1C excretion during follicular phase days 2-5 was lower (P=0.05) in ExOvul, ExLPD, and ExAnov cycles compared to SedOvul cycles and remained lower (P < 0.02) in the ExLPD and ExAnov cycles during days 6 12. The elevation in FSH during the luteal-follicular transition was lower (P < 0.007) in ExLPD (0.7+/-0.1 ng/mg Cr) cycles compared to SedOvul and ExOvul cycles (1.0+/-0.1 and 1.1+/-0.1 ng/mg Cr, respectively). Energy balance and energy availability were lower (P < 0.05) in ExAnov cycles than in other menstrual cycle categories. The blunted elevation in FSH during the luteal-follicular transition in exercising women with LPD may explain their lower follicular estradiol levels. These alterations in FSH may act in concert with disrupted LH pulsatility as a primary and proximate factor in the high frequency of luteal phase and ovulatory disturbances in regularly menstruating, exercising women. PMID- 9851756 TI - The role of cytotoxic chemotherapy in the management of aggressive and malignant pituitary tumors. AB - Pituitary tumors are mostly benign lesions, although 5-35% are locally invasive. A small number exhibit a more aggressive course, infiltrating dura, bone and sinuses, and are designated highly aggressive. However, the presence of metastases separate from the pituitary in the central nervous system or at a distance is necessary to designate pituitary tumors as carcinomas, i.e. truly malignant. When conventional therapeutic modalities fail, systemic chemotherapy remains the last option. We report seven such patients, three with highly aggressive and four with malignant pituitary tumors (n=4) four women; median age, 32 yr; range, 23-48 yr), who received one or more courses of chemotherapy with lomustine and 5-fluorouracil (median, two courses; range, one to six courses). Three patients with systemic metastatic disease had a shorter survival (median, 5 months; range, 1-14 months) than the one patient with central nervous system metastases alone (10 yr). A patient with an aggressive nonmetastatic prolactinoma who initially responded to chemotherapy died from another nondisease-associated cause. Two patients, one with an aggressive and one with a metastatic tumor, achieved symptomatic improvement with a median duration of 6 months. A hormonal reduction greater than 50% was observed in two of seven patients; only one patient who had an aggressive tumor obtained an objective tumor response. The median survival from the time of initiation of chemotherapy in patients with malignant tumors ranged from 3-65 months. Two patients with malignant tumors developed disease progression while receiving chemotherapy; no patient with extracranial metastases showed a response. Treatment was well tolerated, with minimal individual side-effects. Three patients with no response to initial treatment received different chemotherapeutic regimens with no additional response. All patients with metastatic malignant tumors eventually died. Treatment with cytotoxic chemotherapy is noncurative, and current experience is limited. Until another more specific form of treatment is available, chemotherapy may still be of some value in patients with highly aggressive and malignant pituitary tumors, at least in achieving a temporary remission or delay in progression. The combination of lomustine/5-fluorouracil proved easy to administer with minimal toxicity, although the response rate was only 14%. Until a more specific treatment is found, an optimal chemotherapeutic regimen needs to be established. PMID- 9851757 TI - Familial clustering of insulin and abdominal visceral fat: the HERITAGE Family Study. AB - Abdominal visceral fat (AVF) is an obesity-related phenotype thought to be associated with insulin resistance, diabetes mellitus, and atherosclerosis. Significant genetic influences on both AVF and insulin levels have been reported. However, information is lacking as to whether common genetic influences on AVF and insulin levels exist. AVF was assessed by computed tomography scan, and fasting insulin was measured by RIA in 512 members of 98 sedentary Caucasian families participating in the HERITAGE Family Study. Baseline data, collected before exercise training, were used in the present investigation. A bivariate familial correlation model was applied to evaluate whether there are familial influences that are common to insulin and AVF before and after adjustment for total fat mass (FM), and to assess the overall heritability of insulin and AVF. The maximal heritability for AVF, before and after adjustment for total FM, was 42% and 50%, respectively; and for insulin, it was 21%. Interestingly, 29% of the familial influences on insulin were also common to AVF, whereas 14% of the familial influences on AVF were shared by insulin. Furthermore, after AVF was adjusted for total FM, these common familial influences were increased to 48% and 20%. Genes and/or familial nongenetic factors with pleiotropic effects seem to influence both AVF and plasma insulin levels to a certain degree. Genes involved in the regulation of lipid storage and mobilization in the abdominal fat depot are potential candidates for these genetic pleiotropic effects. PMID- 9851758 TI - Additive effects of the mutations in the beta3-adrenergic receptor and uncoupling protein-1 genes on weight loss and weight maintenance in Finnish women. AB - This study examined whether the Trp64Arg mutation in the beta3-adrenergic receptor (beta3AR) and the A-->G mutation in the uncoupling protein-1 (UCP-1) genes have associations with weight loss and subsequent weight maintenance. Seventy-seven obese (body mass index range, 29-46 kg/m2), clinically healthy, premenopausal women were studied. A 12-wk weight reduction by very low calorie diet (VLCD) was followed by a 40-wk weight maintenance phase. The subjects were divided into four groups according to their beta3AR and UCP-1 genotype: no mutation (control; n=37), only Trp64Arg mutation in the beta3AR gene (n=12), only A-->G mutation in the UCP-1 gene (n=23), and both mutations (n=5). Subjects with both mutations had a lower weight reduction during VLCD than the controls [ 10.5+/-0.6 (+/-SEM) vs. -14.0+/-0.5 kg; P=0.051, by ANOVA]. During the maintenance phase, weight in subjects with both mutations increased by 5.8+/-1.5 kg, but remained unchanged in the controls (-0.5+/-0.8 kg; P=0.041). The changes in weight in subjects with only one of the mutation were close to the results in the controls. Resting energy expenditure, adjusted for fat mass, fat-free mass, and maximal aerobic power, did not change differently between the groups throughout the study. The results suggest that a combination of the Trp64Arg mutation in the beta3AR and the A-->G mutation in the UCP-1 genes may be associated with faster weight gain after a VLCD. PMID- 9851759 TI - Effects of androgen administration on the growth hormone-insulin-like growth factor I axis in men with acquired immunodeficiency syndrome wasting. AB - It is unknown whether hypogonadism contributes to decreased insulin-like growth factor I (IGF-I) production and/or how testosterone administration may effect the GH-IGF-I axis in human immunodeficiency virus (HIV)-infected men with the acquired immunodeficiency syndrome (AIDS) wasting syndrome (AWS). In this study, we investigate the GH-IGF-I axis in men with the AWS and determine the effects of testosterone on GH secretory dynamics, pulse characteristics determined from overnight frequent sampling, arginine stimulation, and total and free IGF-I levels. Baseline GH-IGF-I parameters in hypogonadal men with AWS (n=51) were compared before testosterone administration (300 mg, im, every 3 weeks vs. placebo for 6 months) with cross-sectional data obtained in two age-matched control groups: eugonadal men with AIDS wasting (n=10) and healthy age-matched normal men (n=15). The changes in GH-IGF-I parameters were then compared prospectively in testosterone- and placebo-treated patients. Mean overnight GH levels [1.8+/-0.3 and 2.4+/-0.3 vs. 0.90+/-0.1 microg/L (P=0.04 and P=0.003 vs. healthy controls)] and pulse frequency [0.35+/-0.06 and 0.37+/-0.02 vs. 0.22+/ 0.03 pulses/h (P=0.06 and P=0.002 vs. healthy controls)] were comparably elevated in the eugonadal and hypogonadal HIV-positive groups, respectively, compared to those in the healthy control group. No significant differences in pulse amplitude, interpulse interval, or maximal GH stimulation to arginine administration (0.5 g/kg, i.v.) were seen between either the eugonadal and hypogonadal HIV-positive or healthy control patients. In contrast, IGF-I levels were comparably decreased in both HIV-positive groups compared to the healthy control group [143+/-16 and 165+/-14 vs. 216+/-14 microg/L (P=0.004 and P=0.02 vs. healthy controls)]. At baseline, before treatment with testosterone, overnight GH levels were inversely correlated with IGF-I (r=-0.42; P=0.003), percent ideal body weight (r=-0.36; P=0.012), albumin (r=-0.37; P=0.012), and fat mass (r=-0.52; P=0.0002), whereas IGF-I levels correlated with free testosterone (r=0.35; P=0.011) and caloric intake (r=0.32; P= 0.023) in the hypogonadal HIV positive men. In a stepwise regression model, albumin (P=0.003) and testosterone (P=0.011) were the only significant predictors of GH [mean GH (microg/L)=-1.82 x albumin (g/dL) + 0.003 x total testosterone (microg/L) + 6.5], accounting for 49% of the variation in GH. Mean overnight GH levels decreased significantly in the testosterone-treated patients compared to those in the placebo-treated hypogonadal patients (0.9+/-0.3 vs. 0.2+/-0.4 microg/L; P=0.020). In contrast, no differences in IGF-I or free IGF-I were observed in response to testosterone administration. The decrement in mean overnight GH in response to testosterone treatment was inversely associated with increased fat-free mass (r=-0.49; P= 0.024), which was the only significant variable in a stepwise regression model for change in GH [change in mean GH (microg/L)=-0.197 x kg fat-free mass - 0.53] and accounted for 27% of the variation in the change in GH. In this study, we demonstrate increased basal GH secretion and pulse frequency in association with reduced IGF-I concentrations, consistent with GH resistance, among both hypogonadal and eugonadal men with AIDS wasting. Testosterone administration decreases GH in hypogonadal men with AIDS wasting. The change in GH is best predicted by and is inversely related to the magnitude of the change in lean body mass in response to testosterone administration. These data demonstrate that among hypogonadal men with the AWS, testosterone administration has a significant effect on the GH axis. PMID- 9851760 TI - Association between insulin-like growth factor I and bone mineral density in older women and men: the Framingham Heart Study. AB - Few studies of the GH axis and bone have focused specifically on elderly people. The objective of this study was to determine the association between insulin-like growth factor I (IGF-I) and bone mineral density (BMD) in 425 women and 257 men aged 72-94 who participated in the Framingham Osteoporosis Study component of the Framingham Heart Study in 1992-1993. Serum IGF-I level was determined by RIA. BMD at three femoral sites and the lumbar spine was determined by dual x-ray absorptiometry, and at the radius by single-photon absorptiometry. IGF-I level was positively associated with BMD at all five sites (Ward's area, femoral neck, trochanter, radius, and lumbar spine) in women after adjustment for weight loss and other factors (P < or = 0.01) and protein intake in a subset of participants (0.006 < P < 0.07). A threshold effect of higher BMD was evident at each of the 3 femoral sites and the spine (P < 0.03) but not at the radius for women in the highest quintile of IGF-I (> or = 179 g/liter) vs. those in the lowest four quintiles. IGF-I was not significantly associated with BMD in men. These results indicate that higher IGF-I levels are associated with greater BMD in very old women, and suggest that future clinical trials employing GH may have a role in the development of treatments for older women with osteoporosis. PMID- 9851761 TI - Cortisol secretion is related to electroencephalographic alertness in human subjects during daytime wakefulness. AB - To determine whether human hypothalamo-pituitary-adrenal axis activity is related to the alertness level during wakefulness, 10 healthy young men were studied under resting conditions in the daytime (0900-1800 h) after an 8-h nighttime sleep (2300-0700 h). A serial 70-sec gaze fixation task was required every 10 min throughout the daytime experimental session. The corresponding waking electroencephalographic (EEG) segments were submitted to quantitative spectral analysis, from which EEG beta activity (absolute power density in the 13-35 Hz frequency band), an index of central alertness, was computed. Blood was collected continuously through an indwelling venous catheter and sampled at 10-min intervals. Plasma cortisol concentrations were measured by RIA, and the corresponding secretory rates were determined by a deconvolution procedure. Analysis of individual profiles demonstrated a declining tendency for EEG beta activity and cortisol secretory rate, with an overall temporal relationship indicated by positive and significant cross-correlation coefficients between the two variables in all subjects (average r=0.565, P < 0.001). Changes in cortisol secretion lagged behind fluctuations in EEG beta activity, with an average delay of 10 min for all the subjects. On the average, 4.6+/-0.4 cortisol secretory pulses and 4.9+/-0.5 peaks in EEG beta activity were identified by a detection algorithm. A significant, although not systematic, association between the episodes in the two variables was found: 44% of the peaks in EEG beta activity (relative amplitude, near 125%; P < 0.001) occurred during an ascending phase of cortisol secretion, cortisol secretory rates increasing by 40% (P < 0.01) 10-min after peaks in EEG beta activity. However, no significant change in EEG beta activity was observed during the period from 50 min before to 50 min after pulses in cortisol secretion. In conclusion, the present study describes a temporal coupling between cortisol release and central alertness, as reflected in the waking EEG beta activity. These findings suggest the existence of connections between the mechanisms involved in the control of hypothalamo-pituitary-adrenal activity and the activation processes of the brain, which undergoes varying degrees of alertness throughout daytime wakefulness. PMID- 9851762 TI - Body weight and body composition changes after treatment of hyperthyroidism. AB - Body composition changes in nine adults with hyperthyroidism were determined with dual energy x-ray absorptiometry and computed tomography at diagnosis and after 3 and 12 months of euthyroidism achieved by surgery, antithyroid drugs, or treatment with radioiodine. Mean body weight was 67.6 kg at diagnosis and increased 2.7 kg (P=0.06) and 8.7 kg (P < 0.001) after 3 and 12 months of euthyroidism, respectively. Basal metabolic rate decreased from 2087 Cal/24 h at diagnosis to 1601 Cal/24 h at 12 months (P=0.001), whereas reported energy intake dropped from 3244 to 2436 Cal/24 h (P=0.01). According to dual energy x-ray absorptiometry, body fat was unchanged at 3 months, but increased by 5.3 kg (P < 0.0001) at 12 months. Fat-free mass increased 2.7 kg (P=0.003) at 3 months and 3.5 kg (P < 0.0001) at 12 months. Changes in bone mineral content and density did not reach significance. According to computed tomography, skeletal muscle plus skin areas increased by 11% (trunk) and 18% (thigh) at 3 months and by 17% (trunk) and 25% (thigh) at 12 months. There was no increase in sc adipose tissue (AT) at 3 months, but at 12 months this AT depot increased by 15% (thigh) and 33% (trunk). Intraperitoneal AT showed a borderline significant increase by 28% (P=0.08) at 3 months and by 40% (P=0.015) at 12 months. Areas of visceral organs and bone tissue of femur did not change significantly during the study. It is concluded that during early recovery from hyperthyroidism, priority is given to the replenishment of skeletal muscles and ip AT, whereas sc AT is increased at a later stage. PMID- 9851763 TI - Bone mass in prepubertal children: gender differences and the role of physical activity and sunlight exposure. AB - Retrospective studies have suggested that the prepubertal years may be an important window of opportunity to increase bone mass, but there have been few direct studies and little exploration of gender differences in this age group. In this study, we report the associations among physical activity measures, sunlight exposure, body composition, and bone mass in 8-yr-old children. We studied 330 children in 1996 (115 girls and 215 boys; response rate, 60%) who had previously taken part in a cohort study of cot death in 1988. They had measurement of anthropometrics (height, weight, and body composition), sunlight exposure (by questionnaire), and physical activity [questionnaire, muscle strength by dynamometry, and bicycle ergometric physical work capacity at a pulse of 170 beats/min (PWC170)]. Bone mineral density (BMD) was assessed at the spine, femoral neck, and total body by a Hologic QDR2000 densitometer. In females only, PWC170 [hip, 2.4%/quartile (95% confidence interval (CI), 0.3-4.5); spine, 1.7%/quartile (95% CI, 0.0-3.4); size adjusted] was associated with BMD, whereas in males only, BMD was associated with both sports participation (hip, 4.2% (95% CI, 1.1-7.3); spine, 4.3% (95% CI, 0.9-7.7)] and muscle strength [hip, 1.7%/quartile (95% CI, 0.0-3.4); but not spine; size adjusted]. Winter sunlight exposure was associated with BMD in girls [hip, 2.9%/category (95% CI, 0.7-5.0); spine, 3.6%/category (95% CI, 1.4-5.8)], but not in boys [hip, 0.3%/category (95% CI, -1.4 to +2.0); spine, 1.4%/category (95% CI, -0.7 to +3.5)]. Males and females were very similar in body size. However, males had higher size-adjusted BMD at the hip (9.6%; 95% CI, 6.9-14), whereas females had higher size-adjusted BMD at the spine (3.2%; 95% CI, 0.8-5.6%). In conclusion, this study has suggested that physical activity and exposure to sunlight are important in the bone mineralization of prepubertal male and female children. The magnitude of both gender and environmental differences in bone mass in this age group is substantial, suggesting that modification at this stage of life may influence peak bone mass and possibly fracture risk in later life. PMID- 9851764 TI - Normal volumetric bone mineral density and bone turnover in young men with histories of constitutional delay of puberty. AB - It has been suggested that an appropriate timing of puberty is necessary for normal bone mineral density (BMD) acquisition, which may not be achievable in children with constitutional delay of puberty (CDP). To assess the effect of pubertal delay on BMD, we measured areal BMD (aBMD) at lumbar spine, by dual energy x-ray absorptiometry (DEXA), in a group of patients with CDP (n=21; mean age, 21.8+/-1.7 yr) at final height and in healthy controls (n=12; mean age, 19.3+/-1.3 yr). A subset of seven patients (group a) were untreated, whereas six subjects (group b) had received im testosterone depot (100 mg/month, for 6-12 months) and 8 boys (group c) oral oxandrolone (1.25-2.5 mg/daily, for 6-28 months) for their pubertal delay. Volumetric BMD (vBMD) was calculated from DEXA measurements. aBMD was reduced in patients with CDP (1.101+/-0.134 g/cm2), in comparison with controls (1.222+/-0.091 g/cm2; P < 0.009); no significant differences were found among the groups (group a, 1.089+/-0.133 g/cm2; group b, 1.111+/-0.118 g/cm2; group c, 1.103+/-0.160 g/cm2). vBMD was not significantly different in patients with CDP (0.327+/-0.021 g/cm3) and in controls (0.337+/ 0.017 g/cm3; P= not significant); no significant differences were found among the groups (group a, 0.326+/-0.016 g/cm3; group b, 0.332+/-0.022 g/cm3; group c, 0.330+/-0.021 g/cm3). No differences were found in mineral metabolism and in bone markers between patients and controls; patients did not report an increased fracture rate, compared with controls. Our data indicate that: 1) men with CDP have normal vBMD; 2) the reduced aBMD may be the result of uncritical use of DEXA measurements in subjects with altered growth pattern; and 3) androgen administration during pubertal years did not improve BMD in young men with a history of CDP. PMID- 9851765 TI - Relation among left ventricular mass, insulin resistance, and blood pressure in nonobese subjects. AB - Because left ventricular (LV) mass (LVM) is a powerful predictor of future cardiovascular events, it is important to identify hemodynamic and nonhemodynamic factors that increase LVM. We studied the separate contribution to LVM of daily arterial blood pressure (BP) and insulin resistance in a consecutive series of 29 (mean+/-SD age, 43+/-13 yr) nonobese (body mass index, 24+/-1.8 kg/m2), nondiabetic, glucose-tolerant subjects with untreated borderline or mild hypertension. The insulin sensitivity index (SI) was quantitatively determined from the frequently sampled iv glucose tolerance test. BP was characterized by ambulatory 24-h BP monitoring, and LVM index (LVMI) was determined by two dimensional directed M-mode echocardiography. LVMI was directly related to 24-h mean BP (r=0.47; P=0.01). LMVI was also significantly related to Si (r=-0.43; P=0.02). In this nonobese group, neither LVMI nor Si was related to body mass index or age. After adjustment for the influence of BP on LVMI, a significant relation remained between LVMI and Si (P < 0.05). We conclude that in nonobese subjects with high normal BP, insulin sensitivity is related to LVM independently of BP and may be an important modulator of LV growth. In addition to a reduction of arterial BP, optimal prevention of LV hypertrophy in hypertensives may require improved insulin sensitivity. PMID- 9851766 TI - Energy and water metabolism, body composition, and hormonal changes induced by 42 days of enforced inactivity and simulated weightlessness. AB - Inactivity causes profound deleterious changes. We investigated in eight healthy men the impact of a 42-day head-down bed rest (HDBR) on energy and water metabolism and their interrelationships with body composition (BC) and catabolic and anabolic hormones. Total energy expenditure (TEE), total body water, water turnover, and metabolic water formation were assessed by the doubly labeled water method 15 days before and for the last 15 days of HDBR. Resting energy expenditure was determined by indirect calorimetry, and BC was determined by dual energy x-ray absorptiometry. Urinary excretion of cortisol, GH, normetanephrine, metanephrine, urea, and creatinine were measured daily. HDBR resulted in significant reductions in body weight (2%), total body water (5%), metabolic water (17%), and lean body mass (LBM; 4%), but fat mass and water turnover did not change. Segmental BC showed a decreased LBM in legs and trunk, whereas fat mass increased, no significant changes were noted in the arms. The hydration of LBM was unchanged. TEE and energy intake decreased significantly (20% and 13%), whereas resting energy expenditure was maintained. Expenditure for physical activity dropped by 39%. Subjects were in energy balance during HDBR, whereas it was negative during the control period (-1.5 MJ/day). There were decreases in urinary normetanephrine (23%) and metanephrine (23%), but urinary cortisol (28%; weeks 2 and 3), GH (75%; weeks 2-4), and urea (15%; weeks 3 and 4) increased. It was concluded that during prolonged HDBR no relevant modifications in water metabolism were triggered. BC changes occurred in the nonexercised body segments, and the reduction in TEE was due to inactivity, not to LBM loss. Moreover, body weight alone does not accurately reflect the subject's energy state, and energy balance alone could not explain the body weight loss, which involves a transient metabolic stress. PMID- 9851767 TI - Changes in bone mass and bone biomarkers of cynomolgus monkeys during pregnancy and lactation. AB - A substantial amount of calcium is transferred from the mother to the fetus and infant during pregnancy and lactation. Involvement of the skeleton in meeting this demand should be reflected in changes in bone mass and turnover. The purpose of the study was to determine the effects of pregnancy, lactation, and recovery on the skeleton in 43 young (prepeak bone mass) female monkeys. Whole body (WBBMC) and lumbar vertebrae 2-4 bone mineral content were determined by dual x ray absorptiometry at baseline and 1, 4, and 10 months postpartum. Alkaline phosphatase, bone Gla protein, and urinary crosslinks were measured at baseline, during the third trimester, and 1, 4, and 10 months postpartum. Compared to nonpregnant, nonlactating monkeys, pregnant monkeys had similar rates of bone mass gain (nonpregnant, nonlactating WBBMC, 25+/-9 mg/day; pregnant WBBMC, 20+/ 14 mg/day). Compared to pregnant monkeys, lactating females had increased bone turnover, as indicated by elevated bone biomarker levels (lactating alkaline phosphatase, 259+/-20 IU/L) and decreased bone mass (lactating WBBMC, -99+/-21 mg/day). Densitometry showed that bone mass gain in the lactating monkeys did not compensate for lactational loss by 10 months postpartum (WBBMC, 6.95+/-9 mg/day). This lack of recovery may have been due to the fact that serum estrogen concentrations were just beginning to return to baseline at 10 months postpartum. In conclusion, the cynomolgus monkey skeleton responds similarly to that of women during pregnancy and lactation. Recovery from lactational bone loss is not complete by 10 months postpartum. PMID- 9851768 TI - Azoospermia associated with a mutation in the ligand-binding domain of an androgen receptor displaying normal ligand binding, but defective trans activation. AB - Although male infertility affects a significant proportion of couples trying to conceive, the cause of defective spermatogenesis is not known in a large number of cases. Ligand binding studies indicate that a number of these subjects may have defects of the androgen receptor (AR). Genetic screening in subjects with defective spermatogenesis and in 110 fertile controls identified an azoospermic (no sperm in any ejaculates) patient with an amino acid substitution (Gln-->Glu) in residue 798 of the AR gene. This germline mutation was pathogenic because it was not observed in fertile controls, was associated with features of minimal androgen insensitivity in our patient, has been related to more severe grades of androgen insensitivity, and caused a subtle, but significant, decrease in receptor trans-activation function in vitro that is consistent with the phenotype. Despite being located in the middle of the ligand-binding domain of the receptor, the Q798E mutation did not cause any ligand binding defect, indicating that this highly conserved residue has a trans-activation function but does not directly form part of the ligand binding pocket of the receptor. The trans-activation defect of the mutant receptor can be rectified in vitro with the androgenic drug, fluoxymesterone, but not with mesterolone or nortestosterone. Further studies are required to determine the therapeutic relevance of this finding. PMID- 9851769 TI - Mutation of the RET proto-oncogene is correlated with RET immunostaining in subpopulations of cells in sporadic medullary thyroid carcinoma. AB - Mutations in the RET proto-oncogene, which encodes a receptor tyrosine kinase, are associated with the pathogenesis of medullary thyroid carcinoma (MTC). Somatic mutations in RET, predominantly at codon 918, and very rarely at codon 883, have been found in a proportion of sporadic MTC. We have previously shown that approximately 80% of sporadic MTCs had at least one subpopulation with a somatic RET mutation. Uneven distribution of somatic mutation within a single tumor or among metastases from a single individual was notable. In the present study, we sought to correlate RET expression, as demonstrated by RET immunohistochemistry, with mutation status in sporadic MTC for each tumor. Seventy evaluable subpopulations, belonging to 28 unrelated sporadic cases, comprising primary MTC and metastases, were immunostained with two different polyclonal antibodies raised against the C-terminus of RET. The regional presence of codon 918 or 883 seemed to coincide with increased RET immunopositivity in at least 62 of 70 (89%, P < 0.000001) tumor subpopulations. The reasons for this concordance are not entirely clear but could be related to either RNA or protein stability. Preliminary studies have suggested that the presence of somatic codon 918 mutation in MTC has a prognostic significance. If these preliminary results prove true, then given our data, we can further explore the feasibility of RET immunocytochemistry as a rapid assessment for the presence of somatic codon 918 for molecular diagnostic and prognostic purposes. PMID- 9851770 TI - Growth hormone secretagogue receptor expression in human pituitary tumors. AB - The GH secretagogue (GHS) receptor (GHS-R) has been characterized and cloned. It is a member of a family of seven transmembrane receptors and is closely related to the neurotensin and TRH receptors. To determine the expression of this receptor in normal anterior pituitary and in 24 human pituitary adenomas, we analyzed GHS-R messenger ribonucleic acid (mRNA) using a RT-PCR assay. We found that normal human pituitary was positive for the GHS-R signal. In addition, all GH-secreting adenomas and the one TSH-secreting adenoma demonstrated the presence of GHS-R mRNA. Three of four ACTH-secreting tumors and three of nine gonadotroph adenomas were also positive for the GHS-R mRNA. To determine the amounts of GHS-R mRNA in normal pituitary and in representative tumors, semiquantitative competitive PCR was performed. We determined that normal pituitary had approximately 750 molecules/L GHS-R mRNA. The acromegalic tumor had approximately 1.5 x 10(5) molecules/L, and the TSH-secreting tumor had approximately 7.5 x 10(3) molecules/L. Other tumor types contained considerably less, with the ACTH secreting and gonadotroph tumors expressing 7.5 x 10(2) and 3 x 10(2) GHS-R mRNA molecules/L, respectively. These results suggest that GH- and TSH-producing adenomas express GHS-R mRNA at levels 200 and 10 times higher, respectively, than the normal pituitary, and that this receptor expression may be involved in the pathogenesis and growth of these pituitary adenomas. PMID- 9851771 TI - Linkage analysis of candidate genes in autoimmune thyroid disease. III. Detailed analysis of chromosome 14 localizes Graves' disease-1 (GD-1) close to multinodular goiter-1 (MNG-1). International Consortium for the Genetics of Autoimmune Thyroid Disease. AB - The autoimmune thyroid diseases [Graves' and Hashimoto's diseases (GD and HT)] develop in genetically susceptible individuals, but the genes responsible for this susceptibility remain unknown. To identify such genes, we have been testing candidate genes and chromosomal regions using highly polymorphic microsatellite markers. We recently reported evidence for the first locus linked to GD (GD-1) on chromosome 14q31 in a small group of families. We have now extended these studies and analyzed 53 multiplex families with GD and/or HT (323 individuals). Chromosome 14 was screened using 16 microsatellite markers spanning the entire chromosome. Three additional markers located inside candidate genes on chromosome 14 were also studied. Microsatellite markers were amplified using fluorescent labeled primers and separated on an ABI-310 genetic analyzer. The data were analyzed using LIPED software for two-point logarithm of odds (LOD) score analysis and GeneHunter software for multipoint linkage analysis. No linkage of any marker was found to HT or autoimmune thyroid diseases (GD+HT). The previously identified GD-1 locus on 14q31 continued to show evidence of linkage to GD in this much larger set of families. The maximum LOD score was 2.1 obtained for marker D14S81 (theta=0.01), assuming a recessive mode of inheritance and a penetrance of 0.3. Multipoint analysis yielded a maximum LOD score of 2.5 between markers D14S81 and D14S1054. There was no evidence for heterogeneity in our sample. These data again suggest the presence of a major Graves' disease susceptibilitygene (GD-1) on chromosome 14q31. This locus is close to the recently identified multinodular goiter-1 locus. PMID- 9851772 TI - Rapid diagnosis and identification of cross-over sites in patients with glucocorticoid remediable aldosteronism. AB - Glucocorticoid remediable aldosteronism (GRA) is an autosomal dominant cause of primary aldosteronism and high blood pressure resulting from a chimeric 11beta hydroxylase/aldosterone synthase gene. Abnormal expression of aldosterone synthase causes primary aldosteronism, which can be inhibited by glucocorticoids. Diagnosis of GRA has depended on the identification of a restriction enzyme product in genomic DNA of affected individuals. Recently, a two-tube long PCR method was described that allowed diagnosis of GRA in a kindred in Australia. A similar long PCR method confirmed the diagnosis of GRA in members of five northeastern Scotland families previously identified by Southern blotting and detected affected members of five GRA families previously identified in Glasgow. A multiplex PCR protocol is described here that allows the control aldosterone synthase amplification and chimeric gene amplification to be carried out in the same tube. We describe the regions of cross-over in each of 10 kindreds identified in Scotland. To identify cross-over regions in each of the kindreds, the chimeric long PCR product was cloned and sequenced. Five cross-over sites were identified ranging from intron 2 to exon 4, indicating the reliability of the method in identifying chimeric genes resulting from different sites of cross over. PMID- 9851773 TI - Labor-associated changes in interleukin-10 production and its regulation by immunomodulators in human choriodecidua. AB - Parturition is associated with increased production of proinflammatory mediators by gestational tissues. Interleukin-10 (IL-10) is an antiinflammatory cytokine produced by human chorion, decidual, and trophoblast tissues. To study the effects of immunomodulators on IL-10, IL-6, and PGE2 production by human choriodecidua before and after labor, an organ explant system was established. Tissue disks (6 mm) were excised from choriodecidual membranes obtained at term by cesarean section before labor (n=6-7) or after spontaneous vaginal delivery (n=7-8). After 24-h equilibration in medium, the tissues were treated with IL 1beta (10 ng/mL), tumor necrosis factor-alpha (100 ng/mL), lipopolysaccharide (5 microg/mL), dexamethasone (1 micromol/ L), or an appropriate vehicle control (n=3 wells/treatment) for 24 h. Media were harvested, and IL-10, IL-6, and PGE2 concentrations were determined by immunoassay. Basal choriodecidual production rates of IL-10 were significantly decreased with labor (P < 0.001), whereas PGE2 and IL-6 production rates increased. The production of all three substances was increased by IL-1beta, tumor necrosis factor-alpha, and lipopolysaccharide, but inhibited by dexamethasone. In contrast to PGE2 and IL-6, there was significantly increased responsiveness of IL-10 production to inflammatory stimuli after labor, but decreased responsiveness to the inhibitory effects of dexamethasone. These data indicate that IL-10 could play a role in modulating or promoting resolution of the inflammatory processes associated with labor at term and with intrauterine infection-associated preterm labor. PMID- 9851774 TI - The frequency of an inactivating point mutation (566C-->T) of the human follicle stimulating hormone receptor gene in four populations using allele-specific hybridization and time-resolved fluorometry. AB - We have described previously in the Finnish population an inactivating point mutation (566C-->T) in the human FSH receptor (FSHR) gene. In women, this mutation causes hypergonadotropic ovarian failure with arrest of follicular maturation and infertility, whereas in men, there is variable suppression of spermatogenesis, but no absolute infertility. To determine whether the same FSHR mutation occurs in other populations, its frequency was determined in Finland, Switzerland, Denmark, and the Chinese population of Singapore. The mutation was screened for using genomic DNA extracted from whole blood or dried blood spots. Exon 7 of the FSHR gene was first amplified using a pair of biotinylated primers. The PCR products were then immobilized on streptavidin-coated microtitration wells and hybridized using short allele-specific oligonucleotide probes labeled with europium. Time-resolved fluorometry was used for europium signal detection. To test the reliability of this method, 40 isolated DNA samples and 35 dried blood spot samples were blindly tested for the 566C-->T FSHR mutation. The analyses yielded identical results with denaturing gradient gel electrophoresis and allele-specific restriction enzyme digestion of the same samples, thus demonstrating the reliability of the tested method. Automation of this procedure allows the screening of large numbers of samples, which was subsequently carried out to investigate the frequency of the 566C-->T mutation in the study populations. A total of 4981 samples from the above-mentioned 4 countries were analyzed. The frequency of the 566C-->T mutation was 0.96% for all Finnish samples (n=1976), with a strong enrichment of the mutant allele in the northeastern part of the country. Only 1 mutation carrier was identified in the samples from Switzerland (n=1162), whereas none was found in samples from Denmark (n=1094) and the Singapore Chinese (n=540). These results suggest that the 566C- >T mutation of the FSHR gene is enriched in Finland, but is uncommon in other populations. PMID- 9851775 TI - How is whole body protein turnover perturbed in growth hormone-deficient adults? AB - Adult patients with GH deficiency have reduced lean body mass (LBM), muscle mass, and muscle strength, suggesting an underlying abnormality of protein metabolism. As acute GH administration has previously been reported to decrease protein oxidation and increase protein synthesis in GH-deficient (GHD) adults, we investigated whether the converse might occur in untreated GH deficiency by undertaking studies of whole body protein turnover in 10 GHD and 13 normal subjects using a 3-h primed constant infusion of 1-[13C]leucine. Dual energy x ray absorptiometry was used to quantify LBM and fat mass (FM). In normal subjects, LBM was the major, independent determinant of leucine appearance (Ra; r=0.80; P=0.0009), leucine oxidation (r=0.81; P=0.0008), and leucine incorporation into protein (r=0.75; P= 0.003). However, in an analysis of covariance, FM was also a significant independent determinant of leucine Ra (P=0.002) and leucine incorporation into protein (P=0.003). After correcting for LBM and FM, GHD patients had significantly reduced rates of leucine Ra (109.9+/ 4.4 vs. 125.5+/-3.7 micromol/min, respectively; P=0.02) and leucine incorporation into protein (87.0+/-3.9 vs. 100.3+/-3.3 mmol/ min; P=0.02) compared to normal subjects. There was no significant difference in the corrected rates of leucine oxidation between the two groups (22.9+/-1.3 vs. 25.2+/-1.0, GHD vs. normal; P=0.20). In summary, GHD adults have reduced rates of protein synthesis and protein breakdown, but normal rates of irreversible oxidative loss; these findings are discordant with what was predicted from the acute changes in protein metabolism observed with GH administration. We conclude that normalization of protein oxidation may be a homeostatic mechanism that operates to constrain protein loss in GHD adults. PMID- 9851776 TI - Effect of acute pharmacological reduction of plasma free fatty acids on growth hormone (GH) releasing hormone-induced GH secretion in obese adults with and without hypopituitarism. AB - In obesity, there is a markedly decreased GH secretion. The diagnosis of GH deficiency (GHD) in adults is based on peak GH responses to stimulation tests. In the severely obese, peak GH levels after pharmacological stimulation are often in the range that is observed in hypopituitary patients. To distinguish obese subjects from GHD patients, it will be necessary to demonstrate that reduced GH responsiveness to a given test is reversible in the former, but not in the latter, group. Recent studies have shown that reduction of plasma free fatty acids (FFA) with acipimox in obese patients restores their somatotrope responsiveness. There are no data evaluating GH responsiveness to acipimox plus GHRH in obese adults with hypopituitarism. The aim of the present study was to evaluate the effect of acute pharmacological reduction of plasma FFA on GHRH mediated GH secretion in obese normal subjects and obese adults with hypopituitarism. Eight obese patients with a body mass index of 34.2+/-1.2; eight obese adults with hypopituitarism, with a body mass index of 35.5+/-1.9; and six control subjects were studied. All the patients showed an impaired response to an insulin-tolerance test (0.15 U/kg, i.v.), with a peak GH secretion of less than 3 microg/L. Two tests were carried out. On one day, they were given GHRH (100 microg, i.v., 0 min), preceded by placebo; and blood samples were taken every 15 min for 60 min. On the second day, they were given GHRH (100 microg, i.v., 0 min), preceded by acipimox (250 mg, orally, at -270 min and -60 min); and blood samples were taken every 15 min for 60 min. The administration of acipimox induced a FFA reduction during the entire test. Normal control subjects had a mean peak (microg/L) of 23.8+/-4.8 after GHRH-induced GH secretion; previous acipimox administration increased GHRH-induced GH secretion, with a mean peak of 54.7+/-14.5. In obese patients, GHRH-induced GH secretion was markedly reduced, with a mean peak (microg/L) of 3.9+/-1; previous administration of acipimox markedly increased GHRH-mediated GH secretion, with a mean peak of 16.0+/-3.2 (P < 0.05). In obese adults with hypopituitarism, GHRH-induced GH secretion was markedly reduced, with a mean peak (microg/L) of 2+/-0.7; previous acipimox administration did not significantly modify GHRH-mediated GH secretion, with a mean peak of 3.3+/-1.1 (P < 0.05). The GH response of obese patients and obese adults with hypopituitarism was similar after GHRH alone. In contrast, the GH response after GHRH plus acipimox, was markedly decreased in obese adults with hypopituitarism (mean peak, 3.3+/-1.1), compared with obese patients (mean peak, 16.0+/-3.2) (P < 0.05) and control subjects (mean peak, 54.7+/-14.5) (P < 0.01). In conclusion, GH secretion, after GHRH-plus-acipimox administration, is reduced in obese adults with hypopituitarism patients, when compared with obese normal patients. Testing with GHRH plus acipimox is safe and is free from side effects and could be used for the diagnosis of GHD in adults. PMID- 9851777 TI - Characterization of insulin-like growth factor-binding protein-related protein-1 in prostate cells. AB - Insulin-like growth factor-binding protein-related protein-1 (IGFBP-rP1; also known as Mac25, TAF, and PSF) is a member of the IGFBP superfamily. It is a cysteine-rich protein that shares structural and functional similarities with the conventional IGFBPs. In situ hybridization of prostate tissue sections show intense IGFBP-rP1 messenger ribonucleic acid (mRNA) expression in normal stroma and glandular epithelium. There was a significant loss of detectable IGFBP-rP1 mRNA in metastatic prostate tissue. IGFBP-rP1 mRNA (Northern blots) and protein (immunoblots) were detectable in primary cultures ofprostatic stromal and epithelial cells as well as in the immortalized nonmalignant prostatic human epithelial cells, P69, and in the P69 metastatic subline, M12. IGFBP-rP1 expression was not detectable in the prostatic cancer cell lines PC-3, DU145, and LNCaP. IGFBP-rP1 expression was regulated in P69 cells but not in M12 cells. Protein and mRNA expression was up-regulated by IGF-I, transforming growth factor beta, and retinoic acid. The observations that IGFBP-rP1 expression is significantly diminished in prostate tumorigenesis and is regulated in nonmalignant prostate cells suggest IGFBP-rP1 is important in normal prostatic cell growth. PMID- 9851778 TI - Gender differences in basal protein kinetics in young adults. AB - Gender affects energy expenditure and influences the relative utilization of carbohydrate and fat as fuels. However, little is known about the possible effects of gender on protein metabolism. Thus, we compared whole body and plasma (albumin and fibrinogen) protein kinetics in the basal postabsorptive state in young, untrained volunteers divided into two groups according to gender (women: n=17; age, 24+/-4 yr; men: n=17; age, 25+/-2 yr). The two groups were matched for body mass index. Protein kinetics were measured by means of L-[1-14C]leucine infusion. The leucine whole body rate of appearance, an index of proteolysis, and nonoxidative rate of disappearance, an index of protein synthesis, were similar in the two groups. However, the leucine oxidation rate was significantly lower in women compared to men (0.23+/-0.07 vs. 0.31+/-0.08 micromol/kg min; P=0.0062). Similar results were obtained when data were adjusted for estimated body composition. Albumin and fibrinogen fractional secretion rates were not different in the two groups. In conclusion, in the basal state leucine oxidation is lower in women than in men regardless of body composition. This could be one of the factors contributing to the lower metabolic rate in women. PMID- 9851779 TI - Decrease of telomere length in thyroid adenomas without telomerase activity. AB - In somatic cells, telomeres shorten with population doubling, thus limiting their capacity to divide. Telomerase, which synthesizes telomeric repeats, can compensate for such shortening. Telomerase activity is known to be absent from most somatic differentiated cells but is present in germline cells, immortal cell lines, or a large majority of malignant tumors. Autonomous thyroid adenomas are benign tumors composed of highly differentiated cells characterized by TSH independent function and growth. Telomere length and telomerase activity were measured in autonomous and hypofunctioning adenomas and their surrounding tissues. A significant decrease of 3.8+/-1.0 kilobases (kb) was observed in the length of the terminal restriction fragments (TRF) in 12 autonomous adenomas (8.6+/-1.1 kb), compared with the TRF length of their surrounding tissues (12.4+/ 1.6 kb). The same kind of decrease, 3.5+/-1.2 kb, was also observed in 16 hypofunctioning adenomas (12.3+/-1.7 kb in surrounding tissue and 8.8+/-1.6 kb in the adenomas). No telomerase activity was detected either in the 12 autonomous adenomas studied or in most of the quiescent tissues (10 of 12). Most of the hypofunctioning adenomas tested (15 of 16) did not display telomerase activity. These results suggest that the cells have undergone a higher number of cell divisions in the adenomas than in the surrounding tissue. Moreover, there is a larger spread of the TRF length distribution in autonomous adenomas than in the collateral tissue. This could reflect the heterogeneity in proliferation status of the cells in the nodule, some of which have reached the end of their life span, whereas others are still proliferating (but with no malignant potential for the autonomous adenomas). In conclusion, benign adenomas exhibit a shorter and more variable telomere length than the normal collateral quiescent tissue, with no telomerase activity to compensate this loss in telomere length. PMID- 9851780 TI - Human immunodeficiency virus induction of corticotropin in lymphoid cells. AB - Disruption of the linkage among the immune, nervous, and endocrine systems may contribute to the pathology and symptoms of acquired immunodeficiency syndrome (AIDS). We investigated the role of human immunodeficiency virus (HIV) in altering these linkages via induction of corticotropin (ACTH) by lymphocytes. Cultured T lymphocytes (H9 cell line) were infected with HIV-1, after which ACTH production was measured and characterized at various time intervals by immunofluorescence and Western blotting. We report a coordinate expression of ACTH and p24 HIV core protein in H9 cells. Also, the kinetics of HIV-induced ACTH production by H9 T lymphoma cells are demonstrated using three different strains of HIV as well as UV-inactivated HIV. ACTH production corresponded with the appearance of p24 antigen and was maximal 35 days after infection. UV-inactivated HIV and the viral envelope protein, gp120, were also able to induce ACTH production in these cells, indicating that viral replication was not required for the ACTH induction. The HIV-induced ACTH was synthesized de novo and had the size and biological activity of pituitary ACTH. Inhibition of ACTH in HIV-infected lymphocyte cultures by anti-ACTH antiserum enhanced viral p24 expression. The significance of lymphocyte ACTH in AIDS is not clear, but these results suggest that it may restrict HIV replication and possibly infection. PMID- 9851781 TI - Evidence for leptin regulation of food intake in humans. AB - The adipocyte hormone leptin regulates body weight in mice by decreasing food intake and increasing energy expenditure. Whether leptin is of physiological importance for these processes in humans is, however, not clear. We therefore studied the relation between leptin and habitual food intake in 64 healthy postmenopausal women. Dietary habits were assessed with a modified diet history method. Body fat content was measured using bioelectrical impedance. In the 64 women, aged 58.6+/-0.4 yr (mean+/-SD), serum leptin was 19.3+/-12.7 ng/mL, body mass index was 25.0+/-3.5 kg/m2, body fat content was 31.6+/-4.3%, fasting glucose was 4.6+/-0.5 mmol/L, and fasting insulin was 56+/-21 pmol/L. Leptin levels were negatively correlated to total energy intake (r=-0.34; P=0.006), carbohydrate intake (r=-0.36; P=0.004), and total (r=-0.27; P=0.034) as well as saturated fat intake (r=-0.31; P=0.014). Leptin was correlated to the absolute, but not to the percent, intake of these nutrients. When normalized for body fat content, the correlations remained significant. Our results suggest that plasma leptin is involved in the physiological regulation of food intake in humans, and that leptin is related to the quantity rather than the quality of habitual food intake. PMID- 9851782 TI - Activating mutations of the Gs alpha gene are associated with low levels of Gs alpha protein in growth hormone-secreting tumors. AB - Evidence suggests the existence of a direct relationship between cellular Gs alpha content and activation of the adenylyl cyclase system. Data on Gs alpha levels in endocrine tumors that depend on cAMP for growth, particularly pituitary adenomas, are still limited. The levels of Gs alpha protein were evaluated in 11 GH-secreting adenomas with Gs alpha mutations (gsp+) and 15 without (gsp). Complementary DNAs from gsp+ tumors contained very low amounts of wild-type Gs alpha sequences, indicating a preponderance of the mutant Gs alpha transcripts in these tumors. Immunoblotting of Gs alpha protein showed that the two isoforms were present at high levels in all gsp-, but were undetectable or barely detectable in gsp+. The low Gs alpha content in gsp+ tumors was not due to a reduction in ribonucleic acid synthesis or stability, as Gs alpha messenger ribonucleic acid levels were similar in wild-type and mutant tissues. Treatment of gsp- cells with cholera toxin caused a marked reduction of Gs alpha levels. As in other cell systems cholera toxin increases Gs alpha degradation, our data are consistent with an accelerated removal of mutant Gs alpha. This may represent an additional mechanism of feedback response to the constitutive activation of cAMP signaling in pituitary tumors with mutations in the Gs alpha gene. PMID- 9851783 TI - The codon 213 of the 11beta-hydroxysteroid dehydrogenase type 2 gene is a hot spot for mutations in apparent mineralocorticoid excess. AB - In the kidney, the 11beta-hydroxysteroid dehydrogenase type 2 enzyme (11betaHSD2) inactivates glucocorticoids to their inactive ketoforms and thus prevents endogenous glucocorticoids from occupying the nonselective mineralocorticoid receptor in epithelial tissues. Several mutations have been described in the 11betaHSD2 gene in the congenital syndrome of apparent mineralocorticoid excess. These mutations generate partially or completely inactive 11betaHSD2 enzymes. In the present work, we describe an already known mutation in a new patient affected by apparent mineralocorticoid excess, which results in an arginine-to-cysteine mutation (R213C) in the 11betaHSD2 enzyme. This mutation has been found in two other independent families. In vitro expression studies of this mutant provide evidence that the mutant protein is normally expressed, but its activity is abolished. The CGC-to-TGC (C-toT) transition at codon 213 can be considered a typical CpG-consequence mutation. The present finding suggests that the codon R213 of 11betaHSD2 is a hot spot for mutations in this gene, as shown by the occurrence of an R213C point-mutation in several families unrelated to each other. PMID- 9851784 TI - Lactate metabolism of subcutaneous adipose tissue studied by open flow microperfusion. AB - Open flow microperfusion and a novel calibration technique (ionic reference technique) were evaluated for the frequent measurement of the absolute lactate concentration in sc adipose tissue. Furthermore, the influence of the plasma insulin concentration on the lactate concentration of sc adipose tissue was investigated during hyperglycemia. Sixteen lean healthy young men participated in the studies. In the postabsorbtive state the mean sc lactate concentrations were 1.29 and 1.36 mmol/L for the ionic reference technique and the no net flux protocol, respectively (not significant, P > 0.05). The simultaneously measured arterialized plasma lactate concentration was significantly lower at 0.77 mmol/L (P < 0.05). Both the sc lactate concentration (1.8+/-0.33 mmol/L) and the plasma lactate concentration (0.96+/-0.03 mmol/L) were significantly elevated during a hyperinsulinemic euglycemic clamp experiment. During a hyperglycemic clamp experiment the sc lactate concentration reached a significantly elevated plateau (2.15+/-0.27 mmol/L) that was not influenced by the increasing plasma insulin concentration. It is concluded that 1) open flow microperfusion combined with the ionic reference technique enables frequent measurement of the sc lactate concentration; 2) sc adipose tissue is a significant source of lactate release in the postabsorbtive state as well as during hyperinsulinemic clamp conditions; and 3) insulin concentrations greater than 180 pmol/L have no further influence on adipocyte stimulation of sc adipose tissue with respect to lactate release. PMID- 9851785 TI - Magnesium responsiveness to insulin and insulin-like growth factor I in erythrocytes from normotensive and hypertensive subjects. AB - Depletion of intracellular free magnesium (Mg(i)) is a characteristic feature of insulin resistance in essential hypertension, but it is not clear to what extent low Mg(i) levels contribute to insulin resistance, result from it, or both. As insulin-like growth factor I (IGF-I) may improve insulin resistance, we investigated whether this peptide could similarly improve Mg(i) responsiveness to insulin in hypertension, and whether this effect was related to any direct IGF-I effect on Mg(i). 31P-Nuclear magnetic resonance spectroscopy was used to measure Mg(i) in erythrocytes from 13 fasting normotensive and 10 essential hypertensive subjects before and 30, 60, and 120 min after incubation with a physiologically maximal dose of insulin (200 microU/mL) and with different doses of recombinant human IGF-I (0.1-100 nmol/L). In normotensive subjects, IGF-I elevated Mg(i) (P < 0.05) in a dose- and time-dependent fashion, as did insulin (P < 0.05). However, in hypertensive subjects, maximal Mg(i) responses to insulin, but not to IGF-I, were blunted [insulin, 163+/-11 to 177+/-10 micromol/L (P=NS); IGF-I, 164+/-6 to 190+/-11.7 micromol/L (P < 0.05)]. Furthermore, for insulin, but not for IGF-I, cellular Mg(i) responsiveness was closely and directly related to basal Mg(i) levels (insulin: r=0.72; P < 0.01; IGF-I: r=0.18; P=NS). Lastly, blunted Mg(i) responses to insulin could be reversed by preincubation of hypertensive cells with IGF-I. We conclude that 1) both IGF-I and insulin stimulate erythrocyte Mg(i) levels; 2) cellular Mg(i) responses to insulin, but not to IGF-I, depend on basal Mg(i) levels, i.e. the higher the Mg(i) the greater the sensitivity to insulin; and 3) IGF-I potentiates insulin-induced stimulation of Mg(i) at doses that themselves do not raise Mg(i). These effects of IGF-I may underlie at least in part its ability to improve insulin sensitivity clinically. Together, these data support a role for IGF-I in cellular magnesium metabolism and emphasize the importance of magnesium as a determinant of insulin action. PMID- 9851786 TI - The acid-labile subunit of human ternary insulin-like growth factor binding protein complex in serum: hepatosplanchnic release, diurnal variation, circulating concentrations in healthy subjects, and diagnostic use in patients with growth hormone deficiency. AB - Circulating insulin-like growth factor-I (IGF-I) is predominantly bound in the trimeric complex comprised of IGF binding protein-3 (IGFBP-3) and acid-labile subunit (ALS). Circulating concentrations of IGF-I, IGFBP-3 and ALS are believed to reflect the GH secretory status, but the clinical use of ALS determination is not known. We therefore, determined the: 1) hepatosplanchnic release of ALS by liver vein catheterization (n=30); 2) 24-h diurnal variation of ALS (n=8); 3) normal age-related ranges of circulating ALS (n=1158); 4) diagnostic value of ALS in 108 patients with childhood-onset GH deficiency (GHD). We found: 1) no significant arteriovenous gradient over the liver ofALS, IGF-I, and IGFBP-3; 2) the diurnal variation of ALS was 12% (mean coefficient of variation percent); 3) ALS levels increased throughout childhood with maximal levels in puberty, with a subsequent decrease with age in adults; and 4) ALS levels were below -2 SD in 57 of 79 GHD patients (sensitivity 72%) and above 2 SD in 22 of 29 patients with normal GH response (specificity 76%), which was similar, compared with the diagnostic utility of IGF-I and IGFBP-3. Finally, our findings indicate that hepatic ALS production is not measurable by this approach or, alternatively, that the liver is not the primary source of circulating ALS, IGF-I, or IGFBP-3 in humans. In conclusion, we have provided extensive normal data for a novel ALS assay and found that circulating ALS levels exhibit minor diurnal variation. We suggest that ALS determination may be used in future classification of adults suspected of GHD. PMID- 9851787 TI - Molecular genotyping in Brazilian patients with the classical and nonclassical forms of 21-hydroxylase deficiency. AB - The aim of our study was to determine, by allele-specific PCR, the frequency of point mutations in 130 Brazilian patients with the classical and nonclassical forms of 21-hydroxylase deficiency and to correlate genotype with phenotype. The most frequent mutations were 12 splice (41.8% in salt wasting), I172N (32.6% in simple virilizing), and V281L (40.2% in late onset form). The frequency of the 9 most common point mutations was similar to that reported for other countries, except for Del 8 nt and Cluster, which were less frequent in the classical form. Rarer mutations such as P453S, G291S, I7 splice, W405X, R483P, and R483- >frameshift were rarely found or were absent. The 93 fully genotyped patients were classified into 3 mutation groups, based on the degree of enzymatic activity (group A, <2%; group B, approximately 2%, and group C, >18%). In group A, 62% of the cases presented the salt wasting form; in group B, 96% the simple virilizing form; and in group C, 88% the late onset form. We diagnosed 80% of the affected alleles after screening for large rearrangements and 15 point mutations. The absence of previously described mutations in 20% of the affected alleles suggests the presence of new mutations in our population. PMID- 9851788 TI - Effects of methyltestosterone on insulin secretion and sensitivity in women. AB - The frequent coexistence of hyperandrogenism and insulin resistance is well established; however, whether a cause and effect relationship exists remains to be established. In this study we tested the hypothesis that short-term androgen administered to women would induce insulin resistance. To test this hypothesis, regularly menstruating, nonobese women were studied before and during methyltestosterone administration (5 mg tid for 10-12 days) by the hyperglycemic (n=8) and euglycemic, hyperinsulinemic (n=7) clamp techniques. Short-term methyltestosterone administration had no significant effects on the fasting levels of glucose, insulin, c-peptide, glucagon, or glucose turnover. During the hyperglycemic clamp studies, the mean glucose level during the final hour was 203+/-2 and 201+/-1 mg/dL in the methyltestosterone and control studies, respectively. The insulin response to this hyperglycemic challenge was slightly but not significantly greater during methyltestosterone treatment (first phase 59+/-8 vs. 50+/-8 microU/mL in controls; second phase 74+/-9 vs. 67+/-9 microU/mL in controls; total insulin response 133+/-16 vs. 117+/-15 microU/mL in controls). In spite of this, glucose uptake was reduced from the control study value of 10.96+/-1.11 to 7.3+/-0.70 mg/kg/min by methyltestosterone (P < 0.05). The ratio of glucose uptake per unit of insulin was also significantly reduced from a control study value of 14.3+/-1.4 to 9.4+/-1.3 mg/kg/min per microU/mL x 100 during methyltestosterone administration. In the euglycemic hyperinsulinemic clamp studies, insulin was infused at rates of 0.25 and 1.0 mU/kg/min to achieve insulin levels of approximately 25 and 68 microU/mL, respectively. During low dose insulin infusion, rates of endogenous hepatic glucose production were equivalently suppressed from basal values of 2.37+/-0.29 and 2.40+/-0.27 mg/kg/min to 0.88+/-0.25 and 0.77+/-0.26 mg/kg/min in the methyltestesterone and control studies respectively. Whole body glucose uptake during low-dose insulin infusion was minimally affected. During the high-dose insulin infusion, endogenous hepatic glucose production was nearly totally suppressed in both groups. However, whole body glucose uptake was reduced from the control value of 6.11+/-0.49 mg/kg/min to 4.93+/-0.44 mg/kg/min during methyltestosterone administration (P < 0.05). Our data demonstrate that androgen excess leads to the development of insulin resistance during both hyperglycemic and euglycemic hyperinsulinemia. These findings provide direct evidence for a relationship between hyperandrogenemia and insulin resistance, and its associated risk factors for cardiovascular disease. PMID- 9851789 TI - Dual regulation of insulin-like growth factor binding protein-1 levels by insulin and cortisol during fasting. AB - Insulin-like growth factor (IGF) binding protein-1 (IGFBP-1) gene transcription is known to be inhibited by insulin in vivo and in vitro. Levels of IGFBP-1 typically rise during fasting but also rise after acute hypoglycemia, including that induced by insulin, through an unknown mechanism that may involve counterregulatory hormones such as cortisol. To study the regulation of IGFBP-1 secretion during fasting, we measured IGFBP-1, insulin, cortisol, GH, and glucose during the course of standardized fasting studies in a total of 21 children. The fasting studies lasted 13-32 h and were terminated for a whole-blood glucose concentration of less than 50 mg/dL (2.8 mmol). Of the children studied, 9 children had no disorder, 8 had ketotic hypoglycemia, 2 had isolated GH deficiency, and 2 had fatty acid oxidation disorders. During fasting, IGFBP-1 rose above the mean baseline levels of 28+/-5 ng/mL to a mean level+/-SEM of 336+/-59 ng/mL at the time of hypoglycemia (P=0.001). IGFBP-1 was strongly associated with serum insulin and cortisol levels over the entire course of fasting (P < 0.0001)). The interaction of the 2 hormones across time was also strongly significant (P < 0.0001). There was no statistically significant association between IGFBP-1 and GH or glucose. At the time of hypoglycemia, insulin levels were suppressed to 1.7 microU/mL or less, and there was no correlation between IGFBP-1 levels at the end of fasting and final insulin level. In contrast, cortisol levels correlated with IGFBP-1 in the final hypoglycemic sample (r=0.56, P < 0.01). Partial correlation analysis revealed that the relationship between IGFBP-1 and cortisol was unchanged when the data was controlled for insulin levels. These data show that insulin and cortisol both regulate IGFBP-1 secretion during fasting; the effects of insulin and cortisol are strong during the course of fasting. Significant hypoglycemia stimulates a further rise in IGFBP-1, which seems to be regulated, in part, by cortisol. The cortisol-induced rise in IGFBP-1 during fasting and during hypoglycemia potentially serves to prevent the hypoglycemic effects of free IGFs. PMID- 9851790 TI - Evidences for an allelic variant of the human LC/CG receptor rather than a gene duplication: functional comparison of wild-type and variant receptors. AB - Two different human LH receptor sequences have been published, differing by a six base pair insertion encoding Leu-Gln at position 55-60. It has recently been proposed that this would reflect the existence of two LH receptor loci in the human genome. The present results demonstrate that both sequences exist as allelic variants in the Caucasian population. Allelic frequency of"LQ variant" and "wild-type" (alphaLQ) allele are 0.26 and 0.74 respectively. In contrast, the LQ allele is virtually absent from the Japanese population. Functional characterization of both alleles by transient expression in COS-7 cells did not reveal any difference between the two receptors, neither for cell surface expression nor for cAMP production and sensitivity to hCG/LH. PMID- 9851791 TI - Molecular diagnosis of residual and recurrent thyroid cancer by amplification of thyroglobulin messenger ribonucleic acid in peripheral blood. AB - Serum thyroglobulin measurement by immunoassay is used to detect residual or recurrent thyroid cancer after thyroid ablation. However, the usefulness of immunoassay is limited by both the requirement for thyroid hormone withdrawal to attain optimal test sensitivity and interference by antithyroglobulin antibodies. To circumvent these problems, we amplified thyroglobulin messenger ribonucleic acid (mRNA) in peripheral blood using RT-PCR and compared the accuracy of this test to serum thyroglobulin immunoassay in patients with thyroid cancer. Thyroglobulin mRNA was amplified from peripheral blood of 77 patients who had undergone thyroidectomy for well differentiated thyroid cancer, 68 of whom while taking thyroid hormone for TSH suppression. Patient staging was based on the most recent radioiodine scan after thyroid hormone withdrawal. Ten normal control subjects were also studied. Among patients taking T4, thyroglobulin mRNA was detected in 26 of 33 patients with either thyroid bed or metastatic iodine-avid tissue on most recent withdrawal scan (79%), whereas serum thyroglobulin was detected in 12 of these 33 patients (36%; P < 0.001). Thyroglobulin mRNA was detected in 7 of 35 patients (20%) with negative radioiodine scans, 12 of 19 patients (63%) with radioiodine uptake in the thyroid bed, and all 14 patients with metastases, including 2 patients with antithyroglobulin antibodies. Thyroglobulin mRNA was detected in all 10 normal subjects. Epithelioid cells that stained strongly with antithyroglobulin antibodies were identified in blood. Detection of circulating thyroglobulin mRNA is a more sensitive marker of residual thyroid tissue or cancer than immunoassay for serum thyroglobulin, particularly in patients treated with thyroid hormone or who have circulating antithyroglobulin antibodies. PMID- 9851792 TI - Progesterone induces calcitonin gene expression in human endometrium within the putative window of implantation. AB - The human endometrium acquires the ability to implant the developing embryo within a specific time window that is thought to open between days 19-24 of the secretory phase of the menstrual cycle. During this period the endometrium undergoes pronounced structural and functional changes induced by the ovarian steroids, estrogen and progesterone, that prepare it to be receptive to invasion by the embryo. The identification of reliable biochemical markers to assess this critical receptive phase in the context of the natural cycle remains one of the major challenges in the study of human reproduction. Our previous studies in a rat model system demonstrated that the expression of calcitonin, a peptide hormone involved in calcium homeostasis, is transiently induced by progesterone in the glandular epithelium at the onset of implantation. Attenuation of calcitonin synthesis in the uterus during the preimplantation phase by administration of calcitonin antisense oligodeoxynucleotides severely impairs implantation of rat embryos, suggesting that this peptide hormone plays a critical role in uterine receptivity. To investigate whether calcitonin is also expressed in the human endometrium during implantation, we monitored the spatio temporal expression of calcitonin on various days of the menstrual cycle. Our studies employing RT-PCR showed that calcitonin messenger ribonucleic acid is expressed in human endometrium during the postovulatory midsecretory phase (days 17-25) of the menstrual cycle, with maximal expression occurring between days 19 21. Very little calcitonin expression was detected in the endometrium in either the preovulatory proliferative (days 5-14) or the late secretory (days 26-28) phase. In situ hybridization and immunocytochemical analyses localized the calcitonin expression predominantly in the glandular epithelial cells of the endometrium. Our studies further showed that calcitonin expression in the human endometrium is under progesterone regulation. Treatment of women with an antiprogestin, mifepristone (RU-486), drastically reduced calcitonin expression in the endometrium. Collectively, these findings reveal that progesterone-induced expression of calcitonin in the secretory endometrium temporally coincides with the putative window of implantation in the human. PMID- 9851793 TI - Different roles of prepubertal and postpubertal germ cells and Sertoli cells in the regulation of serum inhibin B levels. AB - To elucidate the role of germ cells in the regulation of inhibin B secretion, serum inhibin B levels in prepubertal boys and adult men whom had a concurrent testicular biopsy showing either normal or impaired testicular function were compared. In addition, by immunohistochemistry the cellular localization of the two subunits of inhibin B (alpha and betaB) were examined in adult testicular tissue with normal spermatogenesis, spermatogenic arrest, or Sertoli cell only tubules (SCO) as well as in normal testicular tissue from an infant and a prepubertal boy. Adult men with testicular biopsy showing normal spermatogenesis (n=8) or spermatogenic arrest (n=5) had median inhibin B levels of 148 pg/mL (range, 37-463 pg/mL) and 68 pg/mL (range, 29-186 pg/mL), respectively, corresponding to normal or near-normal levels of our reference population (165 and 31-443 pg/mL; n=358). Men with SCO (n=9) had undetectable or barely detectable (n=1) serum levels of inhibin B. In contrast to adults, prepubertal boys with SCO (n=12) all had measurable serum inhibin B levels that corresponded to our previously determined normal range in healthy prepubertal boys (n=114). However, in postpubertal samples from the same SCO boys, inhibin B levels were undetectable as in the adult SCO men. Intense inhibin alpha-subunit immunostaining was evident in Sertoli cells in both prepubertal and adult testes. In the prepubertal testis, positive immunostaining for the betaB-subunit was observed in Sertoli cells. In the adult testis, intense immunostaining for the betaB-subunit was evident in germ cells from the pachytene spermatocyte to early spermatid stages and to a lesser degree in Leydig cells, but not in Sertoli cells or other stages of germ cells. Thus, surprisingly, in adult men the two subunits constituting inhibin B were expressed by different cell types. We speculate that during puberty Sertoli cell maturation induces a change in inhibin subunit expression. Thus, immature Sertoli cells express both alpha and betaB inhibin subunits, whereas fully differentiated Sertoli cells only express the alpha subunit. The correlation in adult men between serum inhibin B levels and spermatogenesis may be due to the fact that inhibin B in adult men is possibly a joint product of Sertoli cells and germ cells, including the stages from pachytene spermatocytes to early spermatids. PMID- 9851794 TI - Expression of Ob receptor in normal human adrenals: differential regulation of adrenocortical and adrenomedullary function by leptin. AB - The major effects of leptin, an adipostatic hormone produced in fat tissue, are exerted through the hypothalamic-pituitary-adrenal axis and the systemic sympathetic/adrenomedullary system at the level of the central nervous system. Here, we examined the direct effects of leptin on the adrenal gland, a peripheral end organ of both the hypothalamic-pituitary-adrenal axis and the sympathetic/adrenomedullary system. As cortical and chromaffin tissues are intermingled in the human adrenal, we employed the novel technique of laser capture microdissection to analyze these systems separately. Functional full length leptin receptor messenger ribonucleic acid and all human isoforms Ob219.1 3 were demonstrated by RT-PCR in both cortical and medullary tissue. Immunohistochemical staining of leptin receptor protein, however, demonstrated a strong signal only in the adrenal cortex, whereas there was weak positive staining in the medulla. Corticotropin (ACTH)-induced adrenal aldosterone, cortisol, and dehydroepiandrosterone secretion was inhibited by leptin in a concentration-dependent manner, whereas this hormone had no significant effect on catecholamine release by primary cultures of human adrenal chromaffin cells. Leptin itself was not expressed in human adrenal tissue, excluding a local paracrine or autocrine function of this peptide. In conclusion, this is the first report identifying functional leptin receptor in human adrenal tissue and showing a differential action of leptin on human adrenocortical and chromaffin hormone production. This peripheral action of leptin on the adrenal gland provides an additional important link between the human stress response and body weight regulation. PMID- 9851795 TI - Human spermatogenesis in vitro: respective effects of follicle-stimulating hormone and testosterone on meiosis, spermiogenesis, and Sertoli cell apoptosis. AB - In spite of the availability of abundant data about in vitro spermatogenesis in laboratory animals, studies on human in vitro spermatogenesis are scarce. This study employed a relatively simple culture system, involving all cell types of seminiferous tubules, to analyze the effects of FSH and testosterone (T) on different characteristics of human germ and Sertoli cells in culture. By using fluorescence in-situ hybridization, we show that in vitro reduction of germ cell ploidy can be stimulated by FSH but not by T. FSH, but not T, also induced unexpectedly rapid (24-48 h) morphological changes resembling spermiogenesis, although individual changes (spermatid nucleus condensation and protrusion, cell body elongation, and flagellar growth) proceeded in an uncoordinated way and mostly resulted in the development of abnormal forms of elongated spermatids. Though ineffective alone, T potentiated the effects of FSH on meiosis and spermiogenesis. These effects of T were probably caused by the prevention of Sertoli cell apoptosis, an effect that could not be mimicked by FSH. These data show that, in the presence of high concentrations of FSH and T, human spermatogenesis can proceed in vitro with an unusual speed, but the resulting gametes are morphologically abnormal. The potential practical relevance of these findings to assisted reproduction remains to be assessed. PMID- 9851796 TI - Deficient 17beta-hydroxysteroid dehydrogenase type 2 expression in endometriosis: failure to metabolize 17beta-estradiol. AB - Aberrant aromatase expression in stromal cells of endometriosis gives rise to conversion of circulating androstenedione to estrone in this tissue, whereas aromatase expression is absent in the eutopic endometrium. In this study, we initially demonstrated by Northern blotting transcripts of the reductive 17beta hydroxysteroid dehydrogenase (17betaHSD) type 1, which catalyzes the conversion of estrone to 17beta-estradiol, in both eutopic endometrium and endometriosis. Thus, it follows that the product of the aromatase reaction, namely estrone, that is weakly estrogenic can be converted to the potent estrogen, 17beta-estradiol, in endometriotic tissues. It was previously demonstrated that progesterone stimulates the inactivation of 17beta-estradiol through conversion to estrone in eutopic endometrial epithelial cells. Subsequently, 17betaHSD type 2 was shown to catalyze this reaction, and its transcripts were detected in the epithelial cell component of the eutopic endometrium in secretory phase. Because 17beta-estradiol plays a critical role in the development and growth of endometriosis, we studied 17betaHSD-2 expression in endometriotic tissues and eutopic endometrium. We demonstrated, by Northern blotting, 17betaHSD-2 messenger ribonucleic acid (RNA) in all RNA samples of secretory eutopic endometrium (n=12) but not in secretory samples of endometriotic lesions (n=10), including paired samples of endometrium and endometriosis obtained simultaneously from eight patients. This messenger RNA was not detectable in any samples of proliferative eutopic endometrium or endometriosis (n=4) as expected. Next, we confirmed these findings by demonstration of immunoreactive 17betaHSD-2 in epithelial cells of secretory eutopic endometrium in 11 of 13 samples employing a monoclonal antibody against 17betaHSD-2, whereas 17betaHSD-2 was absent in paired secretory endometriotic tissues (n=4). Proliferative eutopic endometrial (n=8) and endometriotic (n=4) tissues were both negative for immunoreactive 17betaHSD-2, except for barely detectable levels in 1 eutopic endometrial sample. Finally, we sought to determine whether deficient 17betaHSD-2 expression in endometriotic tissues is due to impaired progesterone action in endometriosis. We determined by immunohistochemistry the expression of progesterone and estrogen receptors in these paired samples of secretory (n=4) and proliferative (n=4) eutopic endometrium and endometriosis, and no differences could be demonstrated. In conclusion, inactivation of 17beta-estradiol is impaired in endometriotic tissues due to deficient expression of 17betaHSD-2, which is normally expressed in eutopic endometrium in response to progesterone. The lack of 17betaHSD-2 expression in endometriosis is not due to alterations in the levels of immunoreactive progesterone or estrogen receptors in this tissue and may be related to an inhibitory aberration in the signaling pathway that regulates 17betaHSD-2 expression. PMID- 9851797 TI - Four contiguous amino acid substitutions, identified in patients with Laron syndrome, differently affect the binding affinity and intracellular trafficking of the growth hormone receptor. AB - We have analyzed the GH receptor (GHR) gene in four individuals with Laron syndrome, and a missense mutation was identified for each patient in the extracellular domain of the GHR (D152H, I153T, Q154P, and V155G). The D152H mutation was previously reported. We have reproduced the three novel mutations in the GHR complementary DNA and analyzed their consequences in human 293 transfected cells. In cells expressing the I153T and V155G mutants, binding of [125I]human GH at the cell surface was very low, whereas binding to total membrane fractions was much less affected, suggesting impaired cell surface expression. Binding assays with cells expressing the Q154P mutant revealed severe defects both at the cell surface and in total particulate membrane fractions. Immunofluorescence experiments confirmed that cell surface expression of the three mutants was altered, and colocalization studies suggested that most of the mutant receptors are retained in the endoplasmic reticulum. Endoglycosidase H resistance tests also indicated that the majority of I153T and V155G GHRs are trapped in the endoplasmic reticulum. Thus, mutations on contiguous amino acids of the GHR result in various defects. The I153T, Q154P, and V155G mutations mainly affect intracellular trafficking and binding affinity of the receptor, whereas the D152H mutation affects receptor expression, dimerization, and signaling. PMID- 9851798 TI - Expression of type 2 11beta-hydroxysteroid dehydrogenase and corticosteroid hormone receptors in early human fetal life. AB - In adult life, the type 2 isozyme of 11beta-hydroxysteroid dehydrogenase (11betaHSD2) protects the mineralocorticoid receptor (MR) from glucocorticoid by inactivating cortisol to cortisone. 11betaHSD2 activity has been reported in human fetal tissues, where glucocorticoids may impair fetal growth yet are also required for normal fetal development. Using digoxigenin-labeled complementary ribonucleic acid (RNA) probes and an in-house 11betaHSD2 antiserum, we have analyzed the expression of 11betaHSD2, MR, and glucocorticoid receptor (GR) in human fetal tissues of gestational age 6-17 weeks (n=15). 11BetaHSD2 expression was absent at gestational age 6+ weeks, but was expressed in abundance in many fetal tissues between 8-12 weeks. At this time, 11betaHSD2 colocalized with GR messenger RNA (mRNA) expression in metanephros, gut, muscle, spinal cord and dorsal root ganglia, periderm, sex chords of testis, and adrenal. In particular within fetal kidney, intense expression of 11betaHSD2 and GR mRNA was observed over Bowman's capsule and the vascular tufts of developing glomeruli as they migrated from the surface of the kidney to the inner cortex. Only lung and adrenal medullary rests demonstrated high levels of GR mRNA but low levels of 11betaHSD2. 11BetaHSD2 mRNA and immunoreactivity staining patterns were similar, with the exception of the fetal adrenal, where mRNA was localized to the outer definitive zone but immunoreactivity was localized to the inner fetal zone. Colocalization of 11betaHSD2 (and GR mRNA) with MR mRNA was observed principally within epithelial cells of collecting ducts, particularly after 16 weeks gestation when the pattern of distribution of 11betaHSD2 became more adult in nature. High levels of MR mRNA were observed within developing bone. The data indicate that 11betaHSD2 in fetal life principally modulates ligand access to the GR in most fetal tissues, notably glomeruli and tubules in the developing kidney, testis, and periderm, and this may be have ramifications for fetal sodium homeostasis and differentiation. The development of tissues previously shown to have a critical requirement for glucocorticoids, such as lung and adrenal medulla, is facilitated by the expression of GR mRNA, but not 11betaHSD2. The expression of MR mRNA in high abundance in bone suggests a role for corticosteroids in human bone development, and the low/absent expression of 11betaHSD2 at this site suggests that it is functionally acting as a GR. PMID- 9851799 TI - Brain opioid receptor measurements by positron emission tomography in normal cycling women: relationship to luteinizing hormone pulsatility and gonadal steroid hormones. AB - The regulation of central mu-opioid receptors in women during the menstrual cycle was explored with positron emission tomography and the selective radiotracer [11C]carfentanil. Ten healthy women were studied twice, during their follicular and luteal phases. Plasma concentrations of estradiol, progesterone, testosterone, and beta-endorphin were determined immediately before scanning. LH pulsatility was measured over the 9 h preceding each of the two positron emission tomography scans. No significant differences in the binding potential of mu opioid receptors (binding capacity/Kd) were observed between phases of the menstrual cycle. However, significant negative correlations were observed between circulating levels of estradiol during the follicular phase and mu-receptor binding measures in the amygdala and hypothalamus, two regions thought to be involved in the regulation of GnRH pulsatility. LH pulse amplitude was positively correlated with mu binding in the amygdala, whereas LH pulse number was negatively correlated with binding in this same region. No significant associations were noted between LH pulse measures and the hypothalamus for this sample. These results suggest that amygdalar mu-opioid receptors exert a modulatory effect on GnRH pulsatility, and that circulating levels of estradiol also regulate central mu-opioid function. PMID- 9851800 TI - The Ala/Val98 polymorphism of the hepatocyte nuclear factor-1alpha gene contributes to the interindividual variation in serum C-peptide response during an oral glucose tolerance test: evidence from studies of 231 glucose-tolerant first degree relatives of type 2 diabetic probands. AB - The third form of maturity-onset diabetes of the young is caused by mutations in the hepatocyte nuclear factor-1alpha gene. Recently, we demonstrated an association between a prevalent polymorphism at codon 98, Ala/Val98, of this gene and a 20% decreased insulin release during an oral glucose tolerance test (OGTT) in middle-aged glucose-tolerant Danish Caucasian subjects. The major objective of the present study was to replicate this finding among glucose-tolerant first degree relatives of type 2 diabetic patients of the same ethnic origin. All participants, 231 glucose-tolerant offspring of 62 type 2 diabetic probands, underwent an OGTT with measurements of plasma glucose, serum insulin, and serum C peptide during the test. Thirty-three heterozygous carriers of the Ala/Val variant were identified, whereas no subjects had the variant in its homozygous form. Ala/Val carriers had a 20% reduction in serum C peptide at 30 min during the OGTT (1225+/-636 vs. 1507+/-624 pmol/L; P=0.02) compared to wild-type carriers. No significant differences in serum insulin levels during the OGTT were observed between carriers of the variant and Ala/Ala homozygotes. In conclusion, among Danish glucose-tolerant first degree relatives of type 2 diabetic patients the Ala/Val98 polymorphism of the hepatocyte nuclear factor-1alpha gene is associated with a decreased serum C-peptide secretion during an OGTT. This finding confirms our previously reported observation of the functional importance of the variant to insulin secretion during an OGTT among middle-aged healthy subjects. PMID- 9851801 TI - Vasodilator actions of urocortin and related peptides in the human perfused placenta in vitro. AB - Urocortin, is a recently isolated peptide belonging to the CRH family that binds with high affinity to the CRH2 receptor. Like CRH, urocortin causes hypotension in the rat, but its vasoactive actions have not yet been studied in the human. We have compared the vasoactive properties of urocortin, CRH, and urotensin-1 in the human fetal placental vasculature in vitro. Single placental lobules were bilaterally perfused (maternal and fetal sides, 5 mL/min each; 95% O2-5% CO2; 37 C), and changes in fetal arterial perfusion pressure were recorded. Submaximal vasoconstriction was induced by PGF2alpha (4+/-0.7 micromol/L), which increased perfusion pressure from 19.6+/-1.4 to 100.7+/-3.1 mm Hg (n=38; P < 0.001). Subsequent fetal arterial infusion of urocortin (0.001-1 nmol/L) caused concentration-dependent vasodilatation. Urocortin was equipotent with urotensin-1 and 25 times more potent than CRH in causing vasodilatation. Nevertheless, the maximum vasodilator responses to each of the peptides were similar (P > 0.05). The CRH receptor antagonist, alpha-helical CRH-(9-41) (0.2 nmol/L) significantly attenuated the vasodilatation produced by urocortin, urotensin-1, and CRH (P < 0.05). These results indicate a possible physiological role for urocortin in the modulation of human fetal placental vascular tone by activation of CRH2-like receptors. PMID- 9851802 TI - Food-dependent Cushing's syndrome: characterization and functional role of gastric inhibitory polypeptide receptor in the adrenals of three patients. AB - In the present work, the presence of gastric inhibitory polypeptide (GIP) receptors and their functional role in the adrenal cells of three patients with food-dependent Cushing's syndrome were studied. RT-PCR and in situ hybridization studies demonstrated the presence of GIP receptor in the adrenals of the three patients. The presence of this receptor was also demonstrated in two human fetal adrenals, but not in two normal adult human adrenals or in the adrenals of one patient with nonfood-dependent Cushing's syndrome. Freshly isolated cells from patient adrenals responded in a dose-dependent manner to the steroidogenic action of both ACTH and GIP, whereas cells from normal adrenals responded only to ACTH. Treatment of cultured normal adrenal cells with ACTH, but not with GIP, increased the messenger ribonucleic acid (mRNA) levels of cholesterol side-chain cleavage cytochrome P-450, P450c17, and 3beta-hydroxysteroid dehydrogenase, whereas both hormones enhanced these mRNAs in patients' adrenal cells, although the effects of ACTH were greater than those of GIP. Moreover, pretreatment with ACTH enhanced the steroidogenic responsiveness of both normal and patient adrenal cells, whereas GIP caused homologous desensitization, and this was associated with a marked reduction of GIP receptor mRNA levels, as demonstrated by RT-PCR and in situ hybridization. Finally, both ACTH and GIP inhibited DNA synthesis in one patient's adrenal cells, whereas in normal adrenal cells only ACTH had this effect. In conclusion, the present data demonstrate that ectopic expression of functional GIP receptors is the main cause of food-dependent Cushing's syndrome. PMID- 9851803 TI - COUP-TFI expression in human adrenocortical adenomas: possible role in steroidogenesis. AB - Chicken ovalbumin upstream promoter-transcription factor I (COUP-TFI) is an orphan nuclear receptor essential for neurogenesis, organogenesis, and cell fate determination. CYP17 gene transcription has recently been shown to be activated by SF-1 (steroidogenic factor-1) binding to a cyclic AMP-responsive sequence within the promoter region of the gene, and inhibited by COUP-TF binding to the sequence. Thus, COUP-TF and SF-1 act as a transcriptional repressor and activator, respectively, of CYP17 gene expression. Transcriptional repression by COUP-TFI is mediated by corepressors, N-CoR (nuclear receptor-corepressor) and SMRT (silencing mediator for retinoid and thyroid hormone-receptor), whereas transcriptional activation by SF-1 is mediated by coactivator SRC-1 (steroid receptor coactivator-1). We therefore examined the expression of COUP-TFI, SF-1, SRC-1, N-CoR, and SMRT in a variety of adrenocortical adenomas and compared the results with CYP17 mRNA levels. We found significantly high COUP-TFI mRNA expression in nonfunctional adenomas (n=8: 220+/-16%; normal 96+/-4%), a deoxycorticosterone-producing adenoma (n=1: 200%), and a pre-clinical Cushing's adenoma (n=1: 280%), intermediate COUP-TFI expression in cortisol-producing adenomas (n=8: 63+/-5%), and low COUP-TFI expression in aldosterone-producing adenomas (n=8: 49+/-4%). In contrast to COUP-TFI, SF-1 mRNA expression did not vary significantly among adrenals. We did not detect the expected negative correlation between COUP-TFI and CYP17 mRNA levels in adrenocortical adenomas. High COUP-TFI expression was associatedwith a nonfunctioning phenotype. Interestingly, the pattern of COUP-TFI expression was similar to the profile of N CoR expression, but not of SMRT expression. These results indicate that COUP-TFI and N-CoR may play a role in steroidogenesis by human adrenocortical adenomas. PMID- 9851804 TI - Systematic mutation screening of the estrogen receptor beta gene in probands of different weight extremes: identification of several genetic variants. AB - Estrogens are known to have an inhibitory effect on food intake in rodents and primates. Decreased estrogen levels that are found for instance in menopausal woman and in ovarectomized rodents result in body weight gain. Estrogen can act both in the periphery and in the central nervous system via at least two different estrogen receptors (alpha and beta). We systematically screened the coding region and part of the 5' and 3'regions of the estrogen receptor beta gene (ER beta) in 96 extremely obese children and adolescents, 50 patients with anorexia nervosa (AN), 28 patients with bulimia nervosa (BN), and 25 healthy underweight individuals. We detected five different sequence variants in the ER beta: a) A 21 bp deletion (codons 238 to 244) was detected in two obese probands and an underweight individual. b) An 846G-->A transition leading to a nonconservative amino acid substitution (G-250-S) was found in two obese male probands. Both a) and b) were located within the flexible hinge region between DNA and ligand binding domain. c) For a 1082G-->A polymorphism we found suggestive evidence for an association between the more common 1082G-allele and anorexia nervosa (nominal p=0.04). d) One silent mutation (1421T-->C) was found solely in two obese probands. e) A common variant is located in the 3' nontranslated region at position 1730(A-->G). We did not detect association of this polymorphism to any of the analyzed phenotypes. We conclude that the ER beta harbors several different mutations and polymorphisms, none of which can readily be associated with the phenotypes under study. PMID- 9851805 TI - A comment on normal intelligence in growth hormone receptor deficiency. PMID- 9851806 TI - Comments on the comparison of low and high dose corticotropin stimulation tests in patients with pituitary disease. PMID- 9851807 TI - Comment on the low-dose corticotropin stimulation test is more sensitive than the high-dose test. PMID- 9851808 TI - Commentary to the article: comparison of low and high dose corticotropin stimulation tests in patients with pituitary disease. PMID- 9851809 TI - Comments on adults with childhood-onset GH deficiency: the psychosocial adjustment can be negatively conditioned by a final "unsatisfactory" height. PMID- 9851810 TI - The effect of number of days in culture and plasminogen activator inhibitor-1 (PAI-1) 4G/5G genotype on PAI-1 antigen release by cultured human umbilical vein endothelial cells. AB - An insertion/deletion (4G/5G) polymorphism in what has been shown to be an enhancer/repressor binding site in the promoter region of the PAI-1 gene has been related to plasma PAI-1 activity. Transfection studies demonstrated increased interleukin-1 stimulated PAI-1 synthesis in cells containing the 4G sequence. To study this response in endothelial cells, first passage HUVEC from 26 umbilical cords were stimulated with interleukin-1 and tumor necrosis factor-alpha. PAI-1 antigen was measured in 24-hour conditioned medium and allele-specific PCR utilized to determine genotype at the 4G/5G locus. Analysis of covariance was used to determine whether the effect of a variable time in culture was masking a difference between genotypes. A trend towards higher PAI-1 levels with increasing time in culture was observed. The geometric mean (95% confidence interval) of the basal rate of PAI-1 release was, 4G/4G 9.7 (7.0, 13.5) ng/24 hours (n=11), 4G/5G 9.5 (6.5, 13.9) ng/24 hours (n=9), and 5G/5G 10.9 (7.8, 15.1) ng/24 hours (n=6). In cells of the same cultures, the interleukin-1 stimulated levels were 25.9 (23.1, 29.1), 27.2 (23.6, 31.3), and 23.1 (19.5, 27.3) ng/24 hours, respectively, corresponding to ratios of stimulated to basal levels of 2.68, 2.87, and 2.12. After adjustment for time in culture the basal PAI-1 release was 4G/4G 10.7, 4G/5G 9.1, and 5G/5G 9.7 ng/24 hours. For interleukin-1 stimulated release the adjusted levels were 26.3, 27.0, and 22.7 ng/24 hours, respectively. Adjusted levels in 4G/4G genotype cells were non-significantly greater than those in cells of 5G/5G genotype by a factor of 1.16 (0.95, 4.08). This study did not demonstrate a significant difference in basal or cytokine stimulated PAI-1 release from cells of different PAI-1 promoter (4G/5G) genotypes but does not exclude increased interleukin-1 stimulated PAI-1 release in the 4G/4G compared with the 5G/5G genotype. PMID- 9851811 TI - Association between polymorphisms in the fibrinogen alpha- and beta-genes on the post-trauma fibrinogen increase. AB - Fibrinogen is an acute phase reactant, and therefore its plasma levels increase after severe injury. Polymorphisms in the fibrinogen alpha and beta genes have been found to be associated with plasma levels of fibrinogen, and it has also been suggested that they are associated with the fibrinogen increase in acute phase situations. In forty-five consecutive patients admitted to the Intensive Care Unit after acute cranial or thoracic trauma, we investigated the influence of four polymorphisms at the fibrinogen loci (-455G/A and BclI (beta gene), TaqI and T/A312 (alpha gene)) on the post-trauma increase of the fibrinogen levels. At admission, fibrinogen levels were comparable in the patients with the different genotypes for the four polymorphisms studied. However, patients carrying the 455A allele of the -455G/A polymorphism had a significantly wider variation and higher peak levels of fibrinogen, during their stay at the intensive care unit, than did the -455GA homozygotes (5.1 g/l (SD 1.3) and 5.9 g/l (SD 1.0), respectively, p<0.05). Such difference was not found for the other studied polymorphisms. The present study suggests that the increase of fibrinogen level in acute phase situations like severe trauma is associated with the beta-gene 455G/A polymorphism. PMID- 9851812 TI - Rat brain capillary thrombomodulin: structure and function. AB - The anticoagulant transmembrane glycoprotein thrombomodulin (TM) is expressed at the luminal surface of vascular endothelial cells. Recently, we showed that TM antigen and TM mRNA are expressed in brain microvessels in several species and that brain capillaries have the capability to activate protein C. The activation of protein C in brain microcirculation was greatly impaired by major stroke risk factors in rats due to downregulation of TM. In this study, a partial sequence of TM was determined from TM mRNA from brain capillaries examined in brain capillaries of the rat, a species that provides a useful model to investigate stroke mechanisms in relation to brain hemostasis. The predicted deduced amino acid sequences for rat TM were compared with other TM sequences. Particularly high homology (77-100%) among functional domains of the protein, i.e., the epidermal growth factor repeats (EGFRs) 1-6 and the transmembrane region, was observed between mice and rats. Somewhat less degree of homology was observed for bovine and human EGFRs 1-6, while the homology of the transmembrane region was 92 96%. All cysteine residues were conserved among the TM sequences, and specific amino acids previously suggested to be essential for activation of protein C by thrombin TM were highly conserved. We conclude that the highly conserved mRNA and protein sequences may reflect a similar anticoagulant role of TM in brain endothelial and systemic vascular endothelial cells across different species. PMID- 9851813 TI - A new system to detect native microaggregates of platelets in vivo, with a novel platelet aggregometer employing laser light scattering. AB - Circulating platelets are susceptible to various stimuli in vessels, and a certain portion of platelets may thereby form native microaggregates (NMAs), which can be visualized by a bother scanning electron microscopy (SEM). We developed an easy method to evaluate the existence and extent of NMAs and the subsequent spontaneous platelet aggregation (SPA) induced by not any agonist but stirring force with a novel platelet aggregometer (PA-200; Kowa, Tsukuba, Japan) employing a particle-counting method based on laser light scattering. In this system, platelet-rich plasma obtained by a delicate centrifugation was infused in a cuvette to replace the content and to avoid stirring shear. An insertion attachment was designed in a coaxial shape to maintain a constant sample volume. Our data indicated that NMAs, tentatively defined as a "small" division by their light scattering (25-400 of the intensity threshold), could be detected by not only a complicated SEM but also by our simple system with the PA-200 in a few minutes. No correlation was found in the comparison of the existence of NMAs and the SPA generation, in aliquot platelets, respectively. These results suggested that NMAs and SPA might be mutually different cluster and have meanings as representations of platelet status. PMID- 9851814 TI - Some effects of prophylactic aspirins and heparins on concurrent arterial and venous thrombosis in the same animal. AB - The antithrombotic effect of unfractionated heparin, a low-molecular weight heparin, oral acetylsalicylic acid, intravenous lysine-aspirin and oral soluble aspirin was measured on platinum wire simultaneously in artery and vein, in groups of 12-20 rats with concurrent untreated controls. Subcutaneous low molecular weight heparin reduced mean thrombus weight by 26% in the artery only. Venous thrombosis was reduced 56% by oral soluble acetylsalicylic acid and 27% by oral aspirin. Other reductions did not reach statistical significance. While thrombus deposition was induced by both 2-cm (upper site) and 1-cm (lower site) wires, the results obtained on the 2-cm wires in the upper vein and artery were more reliable [corrected]. PMID- 9851815 TI - Phosphorylation-dephosphorylation states at different sites affect phosphoprotein phosphatase 1 activity. AB - We have previously reported that the binding of type I collagen to its receptor initiates platelet aggregation involving phosphoprotein phosphatase 1 (PP 1), which coprecipitates with the 65-kDa platelet type I collagen receptor. Phosphorylation of the anti-PP1 precipitation PP1 decreases its enzyme activity. In the present investigation, the mechanism of the decreased enzyme activity was studied by examining the phosphorylation of PP 1 on serine/threonine or tyrosine residues. Phosphoamino acid analysis of the PP 1 indicates that serine, threonine, and tyrosine can all be phosphorylated. We find that the activity of PP 1 decreases with serine/threonine phosphorylation but that phosphorylation of tyrosine residue activates enzyme activity. These results indicate that the activity of platelet phosphoprotein phosphatase 1 is controlled by phosphorylation and dephosphorylation states at multiple, different site(s). PMID- 9851816 TI - A kinetic assay to determine prothrombin binding to membranes. AB - Activation of prothrombin by multisquamase, the prothrombin activator from the venom of Echis multisquamatus (Central Asian sand viper), is inhibited by membranes containing negatively charged anionic phospholipids. This inhibition appears to be due to the fact that the venom activator cannot activate membrane bound prothrombin. Initial steady state rates of prothrombin activation by multisquamase in the presence of phospholipids appeared to depend on the fraction unbound prothrombin only and this phenomenon was used to quantitate binding of prothrombin to membranes of varying phospholipid composition. In this method, the initial rate of prothrombin activation by multisquamase is measured in the absence (total prothrombin) and in the presence of a procoagulant surface (rate depending only on free prothrombin) and from the difference in activation rates the amount of membrane-bound prothrombin is calculated. The validity of the method was established by determination of the binding parameters for prothrombin binding to 100 microM phospholipid vesicles composed of 20 mole% phosphatidylserine and 80 mole% phosphatidylcholine. The binding parameters obtained were Kd=0.84 microM and n=0.021 micromoles prothrombin bound per micromole phospholipid which is in agreement with literature. Due to the nature of the measurement the method is especially suitable to quantitate binding of prothrombin at concentrations as low as 5 nM prothrombin. PMID- 9851817 TI - Polyethyleneglycol-stabilized manganese-substituted hydroxylapatite as a potential contrast agent for magnetic resonance imaging: particle stability in biologic fluids. AB - RATIONALE AND OBJECTIVES: Polymer-stabilized manganese(II)-substituted hydroxylapatite (MnHA) has been investigated as a particulate contrast agent for magnetic resonance imaging. The MnHA core requires a polymer coating to retard opsonization, thereby prolonging its systemic persistence. Therefore, the aim of this study was to assess the stability of various formulations in biologic media in vitro. METHODS: Polyethyleneglycol-coated manganese(II)-substituted hydroxylapatite particles were studied in bovine plasma as a function of the concentration of polymer in the formulation. Particle sizing techniques and nuclear magnetic resonance proton relaxometry were used to evaluate both in vitro and in vivo stability. RESULTS: A small-sized particle (approximately 10 nm diameter) that is stable in bovine plasma and rabbit whole blood was formed in formulations with high amounts of polymer concentration. In formulations with low amounts of polymer concentration, larger-sized particles (approximately 100 nm diameter) were present along with the small-sized population. The larger particles de-aggregated into the small-size particle distribution on dispersion in bovine plasma and rabbit whole blood. CONCLUSIONS: Ultrasmall particles with high surface coat were stable in plasma, whereas larger aggregates de-aggregated. Unlike Mn2+, the interaction of polyethyleneglycol-stabilized manganese(II) substituted hydroxylapatite with plasma proteins was weak. PMID- 9851818 TI - Efficacy of thrombolytic therapy in pulmonary embolism determined by MION enhanced MRA: an experimental study in rabbits. AB - RATIONALE AND OBJECTIVES: This study determined whether contrast-enhanced magnetic resonance angiography could be used as a noninvasive imaging technique to determine the therapeutic effect and endpoint in thrombolysis of acute pulmonary embolism in an animal model. METHODS: New Zealand white rabbits (n = 18) were anesthetized and mechanically ventilated. Single (n = 12 emboli) or dual (n = 12 emboli in 6 animals) pulmonary emboli were created by injecting autologous thrombi through a right internal jugular venous approach. Three dimensional time of flight (TOF) magnetic resonance angiograms were obtained after intravenous administration of 2 mg Fe of a long circulating monocrystalline iron oxide. Animals then received 5000 IU heparin and 1.3 mg recombinant tissue plasminogen activator/kg intravenously, and magnetic resonance angiography was repeated 30 minutes and 60 minutes after initiation of thrombolytic therapy. RESULTS: MION-enhanced magnetic resonance angiography accurately detected pulmonary emboli in all rabbits. Thrombolysis during the observation period was successful in 8 of the 18 animals. In animals with a single embolus, the revascularization rate was 50% (6 of 12 emboli). The rate was 33% (4 of 12 emboli) in animals with multiple emboli. Magnetic resonance angiography allowed determination of thrombus resolution or thrombus persistence. CONCLUSIONS: It was feasible to diagnose pulmonary embolism accurately in this experimental study and to monitor thrombolysis of pulmonary emboli by MION-enhanced magnetic resonance angiography. PMID- 9851819 TI - Prevention of contrast medium-induced renal vasospasm by phosphodiesterase inhibition. AB - RATIONALE AND OBJECTIVES: This study investigated the involvement of cyclic adenosine monophosphate (cAMP) in contrast medium-induced renal vasomotor effects and the efficacy of selective phosphodiesterase (PDE) inhibitors influencing cAMP in preventing contrast medium-induced renal vasospasm. METHODS: Isometric contractions of rabbit renal artery rings were subjected to increasing concentrations of the ionic contrast medium sodium/meglumine diatrizoate (DIA) and the nonionic contrast media iopamidol (IOP) and iodixanol (IOD) and compared with a potassium chloride control. Subsequently increasing concentrations of the nonselective phosphodiesterase inhibitors theophylline and papaverine and the following selective phosphodiesterase inhibitors were applied: vinpocetine, trequinsin, zardaverine, rolipram, and dipyridamole (subtypes I-V) before restimulation of the arterial tissue with contrast medium. RESULTS: Diatrizoate, iopamidol, and iodixanol induced contractions up to 30%, 15%, and 3.5% of the potassium chloride control, respectively. All phosphodiesterase inhibitors markedly inhibited the contrast medium-induced contractions in a dose-dependent manner. The selective phosphodiesterase inhibitors rolipram and trequinsin attenuated these contractions significantly more (92% and 94%) than did zardaverine, dipyridamole, and vinpocetine, with an inhibitory potency of 37%, 41%, and 62%, respectively. CONCLUSIONS: Nonionic contrast media induced renal vasoconstriction less potently than ionic contrast media. Significant differences in the ability to prevent contrast medium-induced vasoconstriction were observed among the various phosphodiesterase subtypes studied. selective phosphodiesterase inhibition with inhibitor subtypes II and IV showed the most promising results in specifically preventing contrast medium-induced renal vasospasm. PMID- 9851820 TI - Direct video-microscopic observation of the dynamic effects of medical ultrasound on ultrasound contrast microspheres. AB - RATIONALE AND OBJECTIVES: Ultrasound can cause destruction of microbubble contrast agents used to enhance medical ultrasound imaging. This study sought to characterize the dynamics of this interaction by direct visual observation of microbubbles during insonification in vitro by a medical ultrasound imaging system. METHODS: Video microscopy was used to observe air-filled sonicated albumin microspheres adsorbed to a solid support during insonation. RESULTS: Deflation was not observed at lowest transmit power settings. At higher intensities, gas left the microparticle gradually, apparently dissolving into the surrounding medium. Deflation was slower for higher microsphere surface densities. Intermittent ultrasound imaging (0.5 Hz refresh rate) caused slower deflation than continuous imaging (33 Hz). CONCLUSIONS: Higher concentrations of microbubbles, lower ultrasound transmit power settings, and intermittent imaging each can reduce the rate of destruction of microspheres resulting from medical ultrasound insonation. PMID- 9851821 TI - Effects of inhaled gases on the ultrasound contrast produced by microspheres containing air or perfluoropropane in anesthetized dogs. AB - RATIONALE AND OBJECTIVES: Inhaled gas mixtures with increased amounts of oxygen cause air containing ultrasound contrast agents to lose efficacy faster than during the inhalation of air. The authors hypothesized that contrast materials containing relatively insoluble gases would decrease the effects of inhaled gases on the ultrasound contrast. METHODS: Anesthetized dogs were ventilated with compressed air and different oxygen/nitrogen gas mixtures. Video densitometric analysis was performed on end diastolic ultrasound images of the heart after administration of Albunex (air-filled microspheres) or Optison (perfluoropropane filled microspheres). RESULTS: Increased concentrations of oxygen caused no change in the contrast intensity produced by Optison in the left ventricular chamber. In the myocardium, however, increases in oxygen caused Optison to produce significantly less enhancement of the myocardial tissue. CONCLUSIONS: The use of perfluoropropane within albumin microspheres prevented the effects of inhaled gas mixtures on contrast produced within the left ventricular chamber. In the myocardium, increased concentrations of oxygen in the inhaled gas mixtures reduce contrast intensity. PMID- 9851822 TI - Binding and lysing of blood clots using MRX-408. AB - RATIONALE AND OBJECTIVES: A thrombus-specific ultrasound contrast agent, MRX-408, has been developed recently. This agent consists of phospholipid-coated microbubbles with a ligand capable of targeting the GPIIb/IIIa receptor, thereby allowing the microbubbles to bind with thrombi rich in activated platelets. In vitro and in vivo animal experiments have been conducted to examine imaging enhancement and sonothrombolysis using this agent compared with a nontargeted agent. METHODS: For clot binding, blood-smeared slides were incubated with microbubbles and examined under a light microscope. Change in backscatter signals from the blood clots after binding was examined by both an ultrasound scanner and two single-element transducers arranged in a transmitter-receiver pair. For clot lysis, either 1-MHz or 20-KHz ultrasound was used to enhance the lysing effects of MRX-408 with or without urokinase. RESULTS: Evidence of binding was demonstrated under a microscope. In vitro experiments showed that the "acoustic signature", or properties, of blood clots changed after binding. Clots became more echogenic and nonlinear. In vivo fundamental ultrasound imaging confirmed that as a result of binding, blood clots were more visible, the area of detection was improved, and shadowing behind clots was more noticeable. Under 1-MHz ultrasound and 30 minutes of treatment, lysis efficiency reached 34% with MRX 408, whereas there was no visible clot lysis with saline. CONCLUSION: The results of these preliminary studies show that as a contrast agent, MRX-408 enhanced clots under ultrasound imaging and facilitated sonothrombolysis with or without thrombolytic drugs. PMID- 9851823 TI - Acoustically active lipospheres containing paclitaxel: a new therapeutic ultrasound contrast agent. AB - RATIONALE AND OBJECTIVES: Paclitaxel-carrying lipospheres (MRX-552) were developed and evaluated as a new ultrasound contrast agent for chemotherapeutic drug delivery. METHODS: Paclitaxel was suspended in soybean oil and added to an aqueous suspension of phospholipids in vials. The headspace of the vials was replaced with perfluorobutane gas; the vials were sealed, and they were agitated at 4200 rpm on a shaking device. The resulting lipospheres containing paclitaxel were studied for concentration, size, acute toxicity in mice, and acoustic activity and drug release with ultrasound. Lipospheres containing sudan black dye were produced to demonstrate the acoustically active liposphere (AAL)-ultrasound release concept. RESULTS: Acoustically active lipospheres containing paclitaxel had a mean particle count of approximately 1 x 10(9) particles per mL and a mean size of 2.9 microns. Acute toxicity studies in mice showed a 10-fold reduction in toxicity for paclitaxel in AALs compared with free paclitaxel. The AALs reflected ultrasound as a contrast agent. Increasing amounts of ultrasound energy selectively ruptured the AALs and released the paclitaxel. CONCLUSIONS: Acoustically active lipospheres represent a new class of acoustically active drug delivery vehicles. Future studies will assess efficacy of AALs for ultrasound mediated drug delivery. PMID- 9851824 TI - Transcutaneous interruption of ultrasound contrast agents for blood flow evaluation. AB - RATIONALE AND OBJECTIVES: The ability to create short boluses in targeted arteries with rapid rise times is limited by the transport of bubbles from the venous to arterial portion of the circulation. Acoustic interruption of contrast agent in arteries may create the short boluses necessary for simple wash-in/wash out measures of blood flow. METHODS: An ultrasound contrast agent was used with spectral Doppler ultrasound to observe contrast interruption in femoral arteries and VX2 carcinoma in a rabbit model. At an upstream location in the femoral artery, single, sinusoidal ultrasound tone bursts at 1.8 MHz with durations of 0.25 to 1 seconds were applied to interrupt the flow of contrast agent injected intravenously. RESULTS: In VX2 carcinoma, bursts as short as 40 cycles produced contrast interruption lasting only one cardiac cycle within the tumor periphery and I(SPPA) <3 W/cm2 produced measurable interruptions. CONCLUSIONS: Acoustic fields applied transcutaneously interrupted flow of contrast agents to form temporally short negative boluses. PMID- 9851825 TI - Changes in magnesium content and subcellular distribution during retinoic acid induced differentiation of HL60 cells. AB - Magnesium (Mg) is required for cellular proliferation; however, the differences in subcellular regulation of Mg between proliferating and differentiated cells has not been determined. We used electron probe microanalysis (EPMA) to investigate the subcellular distribution of Mg in HL60 cells (a promyelocytic leukemia cell line) before and after retinoic acid (RA)-induced differentiation. Most intracellular Mg is bound to ATP and the Mg-ATP complex regulates several metabolic enzymes. We also compared alterations in Mg content following differentiation with the changes in ATP and ADP levels. Using atomic absorption spectrophotometry, we observed a significant decrease (-20%) in cellular Mg content in RA-differentiated HL60 cells. To investigate which intracellular compartments were involved in these changes, we analyzed subcellular elemental composition in freeze-dried cryosections of rapidly frozen undifferentiated and differentiated HL60 cells by EPMA. Following differentiation of HL60 cells, we observed an 18% decrease in Mg content in both the cytoplasm (regions of the cell excluding mitochondria and nuclei) and mitochondria. There was also a significant (40%) decrease in cytoplasmic Ca content after RA-induced differentiation. Nuclear Mg concentration was not significantly different between differentiated and undifferentiated HL60 cells, although differentiation was accompanied by a 30% decrease in the nuclear K/Na ratio. After differentiation, cellular ATP and ADP content decreased by 31 and 40%, respectively. We conclude that during exit from the cell cycle, Mg redistributes within cells and that the decrease in cytoplasmic and mitochondrial Mg is accompanied by a decrease in ATP and ADP content. PMID- 9851826 TI - NADPH oxidase activity and cytochrome b558 content of human Epstein-Barr-virus transformed B lymphocytes correlate with expression of genes encoding components of the oxidase system. AB - We investigated the NADPH oxidase activity, cytochrome b558 content, and gene expression of gp91-phox and p47-phox in normal Epstein-Barr-virus (EBV) transformed B lymphocytes, compared to EBV-transformed B lymphocytes from patients with X-linked chronic granulomatous disease (CGD), normal peripheral blood neutrophils or mononuclear cells, and the A301 or C8166 lymphoblastoid cell lines. CGD phenotypes included both "classic" disease with no detectable gp91 phox protein (termed X91(0)) and "variant" phenotype with reduced but detectable gp91-phox protein (X91(-)). Normal EBV-transformed B lymphocytes show a dose dependent PMA-induced superoxide release. Culturing these cells with IFN-gamma (100 U/ml) and TNF-alpha (1000 U/ml), alone or in combination for 7 days, caused a modest increase in their NADPH oxidase activity (P > 0.05 in all situations). Normal EBV-transformed B lymphocytes have lower NADPH oxidase activity and cytochrome b558 content than peripheral blood neutrophils or mononuclear cells (P < 0.05 in all situations). In contrast, they have higher NADPH oxidase activity and cytochrome b558 content than X91(-) CGD EBV-transformed B lymphocytes (P < 0.05 in all situations). A301 or C8166 lymphoblastoid cell lines and X91(0) CGD EBV-transformed B lymphocytes have barely detectable NADPH oxidase activity or cytochrome b558 content (P < 0.05 in all situations). Gene expression studies also show a modest increase in expression and transcription rates of gp91-phox and p47-phox genes in normal EBV-transformed B cells cultured with IFN-gamma (100 U/ml) and TNF-alpha (1000 U/ml), alone or in combination for 7 days. We conclude that NADPH oxidase activity and cytochrome b558 content correlate with gp91-phox and p47-phox gene expression in EBV-transformed B lymphocytes. PMID- 9851827 TI - Fluorescent labeling of the leukocyte NADPH oxidase subunit p47(phox): evidence for amphiphile-induced conformational changes. AB - The leukocyte NADPH oxidase of neutrophils is a membrane-bound enzyme that catalyzes the production of O-2 from oxygen using NADPH as the electron donor. Dormant in resting neutrophils, the enzyme acquires catalytic activity when the cells are exposed to appropriate stimuli. During activation, the cytosolic oxidase components p47(phox) and p67(phox) migrate to the plasma membrane, where they associate with cytochrome b558, a membrane-integrated flavohemoprotein, to assemble the active oxidase. Oxidase activation can be mimicked in a cell-free system using an anionic amphiphile, such as sodium dodecyl sulfate (SDS) or arachidonic acid, as an activating agent. It has been proposed that conformational changes in the protein structure of cytosolic factor p47(phox) may be an important part of the activation mechanism. The purpose of the present study was to develop an approach to directly monitor conformational changes in p47(phox) when treated with amphiphiles. Cysteines in recombinant p47(phox) were covalently labeled with a sulfhydryl-reactive, environmentally sensitive, fluorescent probe N, N'-dimethyl-N(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1, 3-diazol 4-yl)ethyleneamine (IANBD). A series of mutant p47(phox) proteins in which the individual cysteine (C98, C111, C196, and C378) was replaced with alanine revealed that all four cysteines of p47(phox) are reactive to IANBD. We found that anionic amphiphiles elicited a dose-dependent increase in fluorescence at an emission maximum of 537 nm from IANBD-labeled p47(phox). Furthermore, a blue shift of emission maximum and a decrease in quenching by the ionic quencher, potassium iodide, were observed in the presence of amphiphiles. These results indicate that the amphiphile-mediated increase in fluorescence from IANBD-labeled p47(phox) is due to the conformational change as seen in the leukocyte NADPH oxidase activation. We propose that this alteration in conformation results in the appearance of a binding site through which p47(phox) interacts with cytochrome b558 during the activation process. In addition, recombinant p67(phox) or a peptide containing proline-rich sequence of p22(phox) (residues 149-162) induces the attenuation of the amphiphile-mediated enhancement of fluorescence from IANBD-labeled p47(phox). This supports the notion that both p67(phox) and p22(phox) influence the conformation of p47(phox). PMID- 9851828 TI - Class I heme peroxidases: characterization of soybean ascorbate peroxidase. AB - An efficient expression system [D. A. Dalton et al. Arch. Biochem. Biophys. 328, 1-8, 1996) for soybean nodule ascorbate peroxidase (APX) has, for the first time, been used to generate enzyme in large enough quantities for detailed biophysical analysis. The recombinant APX has been characterized by electronic absorption, EPR, NMR and circular dichroism spectroscopies, and by electrochemistry. Electronic, EPR, and NMR spectra are consistent with a high-spin ferric resting state for the enzyme at 298 K. Low-temperature EPR (7 K) and electronic absorption (77 K) experiments indicate formation of a low-spin heme derivative at these temperatures. The midpoint reduction potential for the Fe(III)/Fe(II) redox couple, determined by spectroelectrochemistry, is -159 +/- 2 mV vs SHE (pH 7.0, 25.0 degrees C, mu = 0.10 M). Circular dichroism spectra of pea and soybean APXs are very similar, indicating common structural features for the two enzymes. The melting temperature of soybean APX, as monitored by circular dichroism spectroscopy, is 49 degrees C. These results represent the first detailed spectroscopic and electrochemical analysis of soybean ascorbate peroxidase and are discussed in the broader context of other class I peroxidases. PMID- 9851829 TI - Interaction of the wheat endosperm lipid-binding protein puroindoline-a with phospholipids. AB - Puroindoline-a is the main component of a new family of proteins that has been suggested to exert an antimicrobial activity in plant seeds through an interaction with lipid membranes. Here the interaction of puroindoline-a with model phospholipid membranes and micelles has been studied using intrinsic tryptophan fluorescence, fluorescence polarization of diphenyl hexatriene, and proteolysis experiments. The protein appears to interact with both zwitterionic and negative phospholipids. The interaction with phosphatidylcholine is characterized by low-affinity surface binding with very limited penetration into the hydrophobic membrane interior. On the other hand, the interaction with phosphatidylglycerol displays a high affinity and involves a partial penetration of the protein into the bilayer interior that disrupts acyl chain packing. The specificity appears to be due to the presence of a stretch of positively charged residues in the protein sequence. In all, the lipid-binding properties of puroindoline-a resemble those of cardiotoxins, another family of proteins for which a disruptive effect on the membrane structure has been involved to explain their biological function. PMID- 9851830 TI - Glyceraldehyde-3-phosphate dehydrogenase inactivation by peroxynitrite. AB - Rabbit muscle glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was inactivated by peroxynitrite under biologically relevant conditions. The decrease of enzymatic activity followed an exponential function, and the concentration of peroxynitrite needed to inactivate 50% of 7 microM GAPDH (IC50) was 17 microM. Hydroxyl radical scavengers did not protect GAPDH from inactivation, but molecules that react directly with peroxynitrite such as cysteine, glutathione, or methionine and the substrate, glyceraldehyde 3-phosphate, afforded significant protection. Assuming simple competition kinetics between scavengers and the enzyme, we estimated a second-order rate constant of (2.5 +/- 0.5) x 10(5) M-1 s-1 at 25 degreesC and pH 7.4 for the GAPDH tetramer. The loss of enzyme activity was accompanied by protein thiol oxidation (two thiols oxidized per subunit) with only one critical thiol responsible of enzyme inactivation. Indeed, the pH profile of inactivation was consistent with the reaction of GAPDH sulfhydryls (GAPDH-SH) with peroxynitrite. Peroxynitrite-inactivated GAPDH was resistant to arsenite reduction and only 15% recovered by 20 mM dithiothreitol, suggesting that GAPDH SH has been mainly oxidized to sulfinic or sulfonic acid, with a minor proportion yielding a disulfide. On the other hand, under anaerobic conditions the peroxynitrite precursor, nitric oxide (*NO), only slowly inactivated GAPDH with a rate constant of 11 M-1 s-1. The remarkable reactivity of the critical thiol group in GAPDH (Cys-149) toward peroxynitrite, which is one order of magnitude higher than that of previously studied sulfhydryls, indicate that it may constitute a preferential intracellular target for peroxynitrite. PMID- 9851831 TI - Effect of NADH-X on cytosolic glycerol-3-phosphate dehydrogenase. AB - At pH 7.05 NADH-X prepared by incubating NADH with glyceraldehyde-3-phosphate dehydrogenase (E.C. 1.2.1.12) was a potent noncompetitive inhibitor, with respect to coenzyme, of NADPH oxidation by pure rabbit muscle cytosolic glycerol-3 phosphate dehydrogenase (E.C. 1.1.1.8) and also a potent inhibitor of NADPH oxidation catalyzed by this enzyme in a rat pancreatic islet cytosolic fraction. It was a much less potent inhibitor of NADPH oxidation catalyzed by this enzyme in a rat liver cytosolic fraction and of NADH oxidation catalyzed by this enzyme from all three sources. Glycerol-3-phosphate dehydrogenase purified from muscle cytosol contains tightly bound NADH-X, NAD, and ADP-ribose, each in amounts of about 0.1 mol per mole of enzyme polypeptide chain. A deproteinized supernatant of this enzyme contained these three ligands and produced the same type of inhibition of the enzyme described above for prepared NADH-X with a Ki, in the reaction with NADPH at pH 7.05, in the range of 0.2 microM with respect to the total concentration of ligands ([ADP-ribose] + [NAD] + [NADH-X] = 0. 2 microM). However, only the NADH-X component could account for the potent inhibition because NAD, ADP-ribose, and the primary acid product (which can be produced from NADH-X) each had a Ki considerably higher than 0.2 microM. Although at pH 7.05 NADH-X inhibited NADPH oxidation considerably more than NADH oxidation, the reverse was the case at pH 7.38. Since the enzyme purified from muscle contains tightly bound NADH-X, NADH-X might become attached to the enzyme in vivo where it could play a role in regulating the ratio of NADH to NADPH oxidation of the enzyme. PMID- 9851832 TI - Protein phosphatase 1 catalytic subunit isoforms from alfalfa: biochemical characterization and cDNA cloning. AB - The catalytic subunit of protein phosphatase 1 (PP1c) was purified from an alfalfa (Medicago sativa) microcallus cell culture. The preparation was inhibited by rabbit muscle inhibitor-2 and okadaic acid and had a molecular mass of 35 kDa. Five distinct cDNAs termed MsPP1alpha, -beta, -gamma, -delta, and -epsilon were cloned from a M. sativa somatic embryo library. MsPP1alpha was identical to a cDNA reported earlier [A. Pay, M. Pirck, L. Bogre, H. Hirt, and E. Heberle-Bors Mol. Gen. Genet. 244, 176-182, 1994], while the others represented novel isoforms encoded by separate genes. The predicted amino acid sequences of MsPP1alpha, beta, -gamma, -delta, and -epsilon were highly similar to each other and to other known PP1c sequences. The GST-MsPP1ss fusion protein expressed in Escherichia coli was catalytically active and was inhibited by inhibitor-2 and okadaic acid. Affinity-purified polyclonal MsPP1antipeptide antibody detected a protein of 36 kDa in crude cell extracts. These results proved that the cDNA clone encoded an active PP1c which was very similar to the purified enzyme. The mRNA and protein concentrations of PP1c as well as the specific activity of protein phosphatase 1 did not change during the cell cycle in a synchronized alfalfa cell culture. On the other hand, the isoforms exhibited different steady-state mRNA levels in different plant organs suggesting tissue-specific functions. PMID- 9851833 TI - Kinetics of Mn3+-oxalate formation and decay in reactions catalyzed by manganese peroxidase of Ceriporiopsis subvermispora. AB - The kinetics of Mn3+-oxalate formation and decay were investigated in reactions catalyzed by manganese peroxidase (MnP) from the basiomycete Ceriporiopsis subvermispora in the absence of externally added hydrogen peroxide. A characteristic lag observed in the formation of this complex was shortened by glyoxylate or catalytic amounts of Mn3+ or hydrogen peroxide. MnP titers had a minor effect on this lag and did not influence the decay rate of the complex. In contrast, Mn2+ and oxalate drastically affected maximal concentrations of the Mn3+-oxalate complex formed, the decay of which was accelerated at high Mn2+ levels. The highest concentration of complex was obtained at pH 4.0, whereas an inverse relationship was found between the pH of the reaction and the decay rate of the complex with MnP present. In the absence of MnP, the best fit for the decay kinetics of the complex gave an order of 3/2 at concentrations in the range of 30-100 microM, with a kobs = 0.012 min-1 M-0.5 at pH 4.0. The rate constant increases at lower pH values and decreases at high oxalate concentrations. The physiological relevance of these findings is discussed. PMID- 9851834 TI - Oligomeric structure and tissue distribution of ficolins from mouse, pig and human. AB - Mouse plasma ficolin was purified by GlcNAc affinity and anion-exchange chromatography. Gel-filtration chromatography and gradient sedimentation indicated that mouse plasma ficolin is a 12-mer of approximately 35 kDa subunits, and electron microscopy showed the same parachute-like structure previously characterized for the pig ficolin 12-mer. Whereas the predominant form in pig plasma is a 24-mer, mouse and human plasma ficolin showed only the 12-mer form. We conclude that mouse plasma ficolin corresponds to the recently described ficolin A. We have identified a second mouse ficolin gene, ficolin B, which means that pig and mouse each have two ficolin genes, and human has three. One ficolin gene in all species is expressed in liver and is the primary source of plasma ficolin. Expression of this gene in other tissues, and expression of the second ficolin gene, appears to vary in different species. PMID- 9851835 TI - 2-Chloro-1,4-dimethoxybenzene cation radical: formation and role in the lignin peroxidase oxidation of anisyl alcohol. AB - 2-Chloro-1,4-dimethoxybenzene (2Cl-1,4-DMB) oxidation by lignin peroxidase (LiP) proceeds via the formation of the 2Cl-1,4-DMB cation radical as indicated by ESR and UV/vis spectroscopy. The products of the LiP-catalyzed oxidation of 2Cl-1,4 DMB were identified as 2-chloro-1,4-benzoquinone and the dimers dichlorotetramethoxybiphenyl and chloro(chlorodimethoxyphenyl)benzoquinone. The addition of anisyl alcohol (AA) rapidly quenched the 2Cl-1,4-DMB cation radical optical absorption bands, suggesting that the cation radical directly mediates the oxidation of AA. When LiP reactions are conducted in the presence of 50 microM 2Cl-1,4-DMB, the enzyme is inactivated; however, this inactivation can be prevented by the addition of AA. This also suggests that the 2Cl-1,4-DMB cation radical formed in the reaction, in turn, oxidizes AA. PMID- 9851838 TI - Cumulative subject index for volumes 349-360 PMID- 9851836 TI - Mechanism of the synergistic induction of CYP2H by isopentanol plus ethanol: comparison to glutethimide and relation to induction of 5-aminolevulinate synthase. AB - We had previously found that combined treatment with isopentanol and ethanol synergistically induced CYP2H protein and activity in cultured chick nepatoytes. Here we investigated the mechanism of induction of CYP2H by the alcohols and whether they caused a coordinate induction of 5-aminolevulinate synthase (ALAS) mRNA. Treatment with isopentanol alone or in combination with ethanol resulted in coordinate increases in CYP2H1 and ALAS mRNAs. With isopentanol alone, the amounts of CYP2H1 and ALAS mRNAs at 4 to 6 h were similar to those observed after treatment with the alcohol combination, but declined by 11 h. Readdition of isopentanol at 11 h again increased the expression of both mRNAs, indicating that the decreases at 11 h were due to limiting amounts of inducer. Similar results were observed in cells exposed to low concentrations of glutethimide. In the combined alcohol treatment, increases in CYP2H1 and ALAS mRNAs were sustained from 4 h to 11 h after addition of the alcohols, but decreased to control levels by 24 h. Using pulse labeling to measure de novo synthesis of CYP2H1/2 protein, we found that the increases in CYP2H1/2 protein reflected the increases in CYP2H1 mRNA. The half-life of CYP2H1/2 protein, measured from pulse-chase experiments, was approximately twofold greater than the half-life of CYP2H1 mRNA. Our results indicate that the alcohols and glutethimide coordinately increase ALAS and CYP2H1 mRNA, and that increases in CYP2H1/2 protein arise from increases in its mRNA. PMID- 9851839 TI - Specification of gut cell fate differs significantly between the nematodes Acrobeloides nanus and caenorhabditis elegans. AB - The classic view of a strictly cell-autonomous development in nematode embryos has been overturned in recent years with the demonstration of various inductive interactions during early development of Caenorhabditis elegans. To examine how conserved the pattern of embryonic cell specification is among nematodes, we have begun to study the pattern in other species after selective elimination of certain early blastomeres. Here we report considerable differences in specification of the gut lineage between C. elegans and Acrobeloides nanus, another free-living soil nematode belonging to the same order. In C. elegans none of the early blastomeres is by itself able to establish a gut lineage for which an inductive interaction between the somatic EMS cell and its germline sister P2 is required. In contrast, in A. nanus all blastomeres of the 3-cell stage carry the potential to generate gut cells. Our data suggest that repressive interactions take place among blastomeres to ensure that under normal conditions only one of them executes the gut fate. Thus, in related species of nematodes with a very conserved morphology, the assignment of cell fate during early embryogenesis appears to involve quite different strategies. PMID- 9851840 TI - A delayed role for nitric oxide-sensitive guanylate cyclases in a migratory population of embryonic neurons. AB - Neuronal differentiation requires a coordinated intracellular response to diverse extracellular stimuli, but the role of specific signaling mechanisms in regulating this process is still poorly understood. Soluble guanylate cyclases (sGCs), which can be stimulated by diffusible free radical gasses such as nitric oxide (NO) and carbon monoxide (CO) to produce the intracellular messenger cGMP, have recently been found to be expressed within a variety of embryonic neurons and implicated in the control of both neuronal motility and differentiation. Using the enteric nervous system (ENS) of the moth, Manduca sexta, we examined the role of NO and NO-sensitive sGCs during the migration and differentiation of an identified set of migratory neurons (the EP cells). Shortly after the onset of their migration, a subset of EP cells began to express NO-sensitive sGC activity (visualized with an anti-cGMP antiserum). Unlike many neurons in the central nervous system, the expression of sGC activity in the EP cells was not transient but persisted throughout subsequent periods of axon elongation and terminal branch formation on the gut musculature. In contrast, nitric oxide synthase activity (visualized using NADPH-diaphorase histochemistry) was undetectable in the vicinity of the EP cells until the period of synapse formation. Manipulations designed to alter sGC and NOS activity in an in vivo embryonic culture preparation had no discernible effect on either the migration or axonal outgrowth of the EP cells. In contrast, inhibition of both of these enzymes resulted in a significant reduction in terminal synaptic branch formation within the postmigratory neurons. These results indicate that while NO-sensitive sGC activity is expressed precociously within the EP cells during their initial migratory dispersal, a role for this signaling pathway can only be demonstrated well after migration is complete, coincident with the formation of mature synaptic connections. PMID- 9851841 TI - The amn gene product is required in extraembryonic tissues for the generation of middle primitive streak derivatives. AB - The primitive streak is the defining feature of the gastrulating mouse embryo. Currently, little is known in the mouse about the mechanisms that mediate the assembly of the primitive streak or about the signaling pathways that specify the different types of mesoderm and endoderm generated from the streak. To gain insight into primitive streak assembly and function, we have conducted a detailed phenotypic characterization of amnionless, a transgene-induced insertional mouse mutation that arrests embryonic development during gastrulation. Our histological and molecular analyses, when examined in the context of the mouse gastrula fate map, lead to the model that middle streak formation is specifically impaired in the amnionless mutant. Significantly, these observations argue that the formation of the middle streak is mediated by a pathway that is genetically separable from those that direct the specification of the distal and proximal streak regions. Intriguingly, our findings from wt ES cell left and right arrow amnionless-/- blastocyst chimeras indicate that this proposed separate pathway for middle streak formation is dependent on amnionless gene functions in the visceral endoderm. PMID- 9851842 TI - Sperm-activating proteins obtained from the herring eggs are homologous to trypsin inhibitors and synthesized in follicle cells. AB - The activation of sperm motility by the egg is an ubiquitous phenomenon in the animal kingdom, but the molecules by which the egg activates sperm motility have been clarified in only a few invertebrate species. In the Pacific herring, Clupea pallasii, mature unfertilized eggs release the sperm-activating proteins which are prerequisite to successful fertilization. Complementary DNA clones encoding herring sperm-activating proteins were isolated from a herring ovarian complementary DNA library and amino acid sequences were deduced. The herring sperm-activating protein(s) is a secretory product(s) with a strong homology to Kazal-type trypsin inhibitors, such as mammalian acrosin inhibitors. The sperm activating proteins were globally distributed in the outermost layer of the egg chorion and its gene was found to be expressed in the follicle cells which surround developing oocytes. These results suggest that in the Pacific herring, trypsin inhibitor-like proteins are synthesized in the follicle cells, secreted, accumulated in the egg chorion during oocyte development, and released into the milieu at spawning to activate the motility of spermatozoa at the time of gamete interaction. PMID- 9851843 TI - Morphogenetic mechanisms of epithelial tubulogenesis: MDCK cell polarity is transiently rearranged without loss of cell-cell contact during scatter factor/hepatocyte growth factor-induced tubulogenesis. AB - Many organ systems are composed of networks of epithelial tubes. Recently, molecules that induce development of epithelial tubules and regulate sites of branching have been identified. However, little is known about the mechanisms regulating cell rearrangements that are necessary for tubule formation. In this study we have used a scatter factor/hepatocyte growth factor-induced model system of MDCK epithelial cell tubulogenesis to analyze the mechanisms of cell rearrangement during tubule development. We examined the dynamics of cell polarity and cell-cell junctions during tubule formation and present evidence for a multistep model of tubulogenesis in which cells rearrange without loss of cell cell contacts and tubule lumens form de novo. A three-dimensional analysis of markers for apical and basolateral membrane subdomains shows that epithelial cell polarity is transiently lost and subsequently regained during tubulogenesis. Furthermore, components of cell-cell junctional complexes undergo profound rearrangements: E-cadherin is randomly distributed around the cell surface, desmoplakins I/II accumulate intracellularly, and the tight junction protein ZO-1 remains localized at sites of cell-cell contact. This suggests that differential regulation of cell-cell junctions is important for the formation of tubules. Therefore, during tubulogenesis, cell-cell adhesive contacts are differentially regulated while the polarity and specialization of plasma membrane subdomains reorganize, enabling cells to remain in contact as they rearrange into new structures. PMID- 9851844 TI - Normal differentiation of cultured lens cells after inhibition of gap junction mediated intercellular communication. AB - The cells of the vertebrate lens are linked to each other by gap junctions, clusters of intercellular channels that mediate the direct transfer of low molecular-weight substances between the cytosols of adjoining cells. Although gap junctions are detectable in the unspecialized epithelial cells that comprise the anterior face of the organ, both their number and size are greatly increased in the secondary fiber cells that differentiate from them at the lens equator. In other organs, gap junctions have been shown to play an important role in tissue development and differentiation. It has been proposed, although not experimentally tested, that this may be true in the lens as well. To investigate the function of gap junctions in the development of the lens, we have examined the effect of the gap junction blocker 18beta-glycyrrhetinic acid (betaGA) on the differentiation of primary cultures (both dissociated cell-derived monolayers and central epithelium explants) of embryonic chick lens epithelial cells. We found that betaGA greatly reduced gap junction-mediated intercellular transfer of Lucifer yellow and biocytin throughout the 8-day culture period. betaGA did not, however, affect the differentiation of these cells into MP28-expressing secondary fibers. Furthermore, inhibition of gap junctions had no apparent effect on either of the two other types of intercellular (adherens and tight) junctions present in the lens. We conclude that the high level of gap junctional intercellular communication characteristic of the lens equator in vivo is not required for secondary fiber formation as assayed in culture. Up-regulation of gap junctions is therefore likely to be a consequence rather than a cause of lens fiber differentiation and may primarily play a role in lens physiology. PMID- 9851845 TI - erbB genes in the mouse uterus: cell-specific signaling by epidermal growth factor (EGF) family of growth factors during implantation. AB - We previously described spatiotemporal expression of various epidermal growth factor (EGF)-like ligands and receptor subtypes, ErbB1 and ErbB2, during the peri implantation period. To better understand the roles of these ligands and their possible signaling schemes in implantation, it is important to define the status of all the ligands and receptor subtypes in the uterus/embryo. No information is available about uterine and embryonic status of ErbB3 or ErbB4 during implantation. We cloned mouse erbB3 and erbB4 cDNAs and examined their expression and bioactivity in the peri-implantation uterus (days 1-8). Two erbB3 (cytoplasmic and extracellular) and three erbB4 (two cytoplasmic and one extracellular) clones were generated. Both forms of the erbB3 clone showed similar transcript profiles, while different transcript profiles were obtained with erbB4 clones. The steady-state levels of erbB3 and erbB4 mRNAs in whole uterine poly(A)+ RNA samples showed little changes during the peri-implantation period, while their unique cell-specific accumulation was noted. erbB3 is predominantly expressed in the epithelial cells, although decidual and embryonic cells also accumulate this mRNA. In contrast, the erbB4 mRNA is primarily expressed in the submyometrial stroma and myometrial connective tissues during this period. Additionally, the extracellular form of the erbB4 clone detected signals in a subpopulation of stromal cells. Autophosphorylation and immunoprecipitation studies provided evidence that uterine ErbB3 and ErbB4 are biologically active. This study provides a comprehensive analysis of possible ligand-receptor signaling schemes for EGF-like ligands in implantation. PMID- 9851846 TI - The cell lineage of a polyclad turbellarian embryo reveals close similarity to coelomate spiralians. AB - Recent molecular evidence suggests the turbellarian Platyhelminthes may represent the extant basal members of the Spiralia and therefore probably exhibit ancient features of the spiralian developmental program. The stereotypic quartet spiral cleavage pattern of the polyclad turbellarian embryo, among other features, indicates that this group may be closely related to the ancestral flatworm; however, polyclad embryos have been the subject of few experimental studies. Here we report the results of a cell lineage analysis of the embryo of the polyclad Hoploplana inquilina based on microinjection of DiI into cleavage-stage blastomeres following formation of each of the four quartets of micromeres. The first quartet gives rise to most of the lateral and anterior ectoderm of the Muller's larva; the second quartet forms largely dorsal and ventral ectoderm as well as the circular muscles; the third quartet forms only small clones of ectoderm; and only the 4d cell of the fourth quartet contributes to larval structure, forming the longitudinal muscles, mesenchyme, and probably endoderm. Our results demonstrate a striking similarity between the cell lineages of polyclad and higher spiralian embryos, in which the four quadrants also bear the same relationships to the larval axes and give rise to comparable larval structures, including derivation of mesoderm from both ectodermal (2b) and endodermal precursors (4d). PMID- 9851847 TI - Lineage and clonal development of gastric glands. AB - Individual gastric glands of the stomach are composed of cells of different phenotypes. These are derived from multipotent progenitor stem cells located at the isthmus region of the gland. Previous cell lineage analyses suggest that gastric glands, as in the colon and small intestine, are invariably monoclonal by adult stages. However, little is known about the ontogenetic progression of glandular clonality in the stomach. To examine this issue, we employed an in situ cell lineage marker in female mice heterozygous for an X-linked transgene. We found that stomach glands commence development as polyclonal units, but by adulthood (6 weeks), the majority progressed to monoclonal units. Our analysis suggests that at least three progenitor cells are required to initiate the development of individual gastric glands if they are analyzed just after birth. Hence, unlike the colon and small intestine, stomachs showed a significant fraction (10-25%) of polyclonal glands at adult stages. We suggest that these glands persist from polyclonal glands present in the embryonic stomach and hypothesize that they represent a subpopulation of glands with larger numbers of self-renewing stem cells. PMID- 9851848 TI - The differential sensitivities of inner ear structures to retinoic acid during development. AB - In order to examine the mechanisms that underlie development of the inner ear, the normal processes were perturbed using all-trans-retinoic acid (RA). By implanting a resin exchange bead saturated with RA into stage 16 (Hamburger and Hamilton, 1951, J. Morphol. 88, 49-92) embryonic day 2.5 chick ears, it was possible to analyze its in vivo effects on inner ear development. There is a temporal window during which the developing chick inner ear is particularly susceptible to the effects of RA (stages 16-19). This RA period of sensitivity precedes evidence of gross morphologic or histologic differentiation by at least 24 h, suggesting that mechanisms controlling formation of key inner ear structures are already in progress. There is a dose dependence on RA, with increasing doses of RA generating increasingly severe phenotypic abnormalities. Data indicate that these effects are due to differential sensitivities of the various inner ear structures to RA during their formation. In general, the vestibular structures were more susceptible to RA effects than the cochlear duct. Furthermore, nonsensory structures such as semicircular canals seemed to display a greater susceptibility to RA than their associated sensory structures (i.e., cristae). Among the three semicircular canals, the superior canal was the most susceptible to RA treatment, whereas the common crus was particularly resistant, suggesting that the molecular mechanisms for each structure's formation are different. The defect in semicircular canal formation is due to problems in the initial outgrowth of the canal plate which in turn is related to a down regulation of early otocyst cell proliferation. This perturbation model provides valuable insight into the processes involved in producing the intricate patterning of the inner ear. PMID- 9851849 TI - Cellularization in Drosophila melanogaster is disrupted by the inhibition of rho activity and the activation of Cdc42 function. AB - Regulation of cytoskeletal dynamics is essential for cell shape change and morphogenesis. Drosophila melanogaster embryos offer a well-defined system for observing alterations in the cytoskeleton during the process of cellularization, a specialized form of cytokinesis. During cellularization, the actomyosin cytoskeleton forms a hexagonal array and drives invagination of the plasma membrane between the nuclei located at the cortex of the syncytial blastoderm. Rho, Rac, and Cdc42 proteins are members of the Rho subfamily of Ras-related G proteins that are involved in the formation and maintenance of the actin cytoskeleton throughout phylogeny and in D. melanogaster. To investigate how Rho subfamily activity affects the cytoskeleton during cellularization stages, embryos were microinjected with C3 exoenzyme from Clostridium botulinum or with wild-type, constitutively active, or dominant negative versions of Rho, Rac, and Cdc42 proteins. C3 exoenzyme ADP-ribosylates and inactivates Rho with high specificity, whereas constitutively active dominant mutations remain in the activated GTP-bound state to activate downstream effectors. Dominant negative mutations likely inhibit endogenous small G protein activity by sequestering exchange factors. Of the 10 agents microinjected, C3 exoenzyme, constitutively active Cdc42, and dominant negative Rho have a specific and indistinguishable effect: the actomyosin cytoskeleton is disrupted, cellularization halts, and embryogenesis arrests. Time-lapse video records of DIC imaged embryos show that nuclei in injected regions move away from the cortex of the embryo, thereby phenocopying injections of cytochalasin or antimyosin. Rhodamine phalloidin staining reveals that the actin-based hexagonal array normally seen during cellularization is disrupted in a dose-dependent fashion. Additionally, DNA stain reveals that nuclei in the microinjected embryos aggregate in regions that correspond to actin disruption. These embryos halt in cellularization and do not proceed to gastrulation. We conclude that Rho activity and Cdc42 regulation are required for cytoskeletal function in actomyosin-driven furrow canal formation and nuclear positioning. PMID- 9851850 TI - p53-mediated germ cell quality control in spermatogenesis. AB - Spontaneous germ cell death is a common cellular process in the mammalian testis, although the function of this process during spermatogenesis is unclear. An investigation was undertaken to determine whether p53 serves as a mechanism in germ cell quality control by causing spontaneous germ cell death. Using an annexin V assay, lower levels of spontaneous apoptosis were found in the testes of p53-/- mice compared to p53+/+ mice. Propidium iodine staining revealed that the greatest reduction in apoptosis and the largest increase in cell numbers occurred in the tetraploid germ cell population of p53-/- mice. Microscopic examination of sperm morphology showed an increased percentage of abnormal forms in p53-/- mice. Furthermore, p53-/- mice sired fewer offspring than p53+/+ mice did when both groups were mated with p53+/+ females. These results suggest that p53 mediates spontaneous testicular germ cell apoptosis and failure to remove defective germ cells by this mechanism results in increased percentages of abnormal sperm and reduced fertility. p53-mediated apoptosis may be an effector of cellular proofreading that acts to maintain the cellular integrity of germ cells during spermatogenesis. PMID- 9851851 TI - Activin-induced factors maintain goosecoid transcription through a paired homeodomain binding site. AB - Previous studies in both Xenopus and zebrafish have shown that goosecoid is one of the first genes to be transcribed at the onset of gastrulation. Goosecoid transcription still initiates when embryos are treated with protein synthesis inhibitors, indicating that it is mediated by preexisting factors and suggesting that goosecoid transcription is immediately downstream of the maternal mesoderm inducing signal. However, goosecoid transcription continues long after this maternal signal has ceased to be active, indicating that there are mechanisms to maintain activin-induced transcription. Our study has focused on understanding the factors required to maintain this transcription. We have defined an element within the zebrafish goosecoid promoter that is sufficient for activin inducibility in both Xenopus and zebrafish embryos. This element, the goosecoid activin element, interacts with two developmentally regulated proteins from Xenopus embryos. A maternal protein interacts through cleavage stages until the midblastula transition, and a second protein binds from the onset of gastrulation. The second protein is zygotically expressed, and its binding is required for activin inducibility in our assay system. We suggest that the zygotic protein we have identified is a good candidate to be involved in the maintenance of goosecoid transcription. Furthermore, this zygotic protein is likely to contain a paired class homeodomain since a consensus binding site for such proteins is present within the goosecoid activin element and is essential for its function. PMID- 9851852 TI - Tinman function is essential for vertebrate heart development: elimination of cardiac differentiation by dominant inhibitory mutants of the tinman-related genes, XNkx2-3 and XNkx2-5. AB - In Drosophila, the tinman gene is absolutely required for development of the dorsal vessel, the insect equivalent of the heart. In vertebrates, the tinman gene is represented by a small family of tinman-related sequences, some of which are expressed during embryonic heart development. At present however, the precise importance of this gene family for vertebrate heart development is unclear. Using the Xenopus embryo, we have employed a dominant inhibitory strategy to interfere with the function of the endogenous tinman-related genes. In these experiments, suppression of tinman gene function can result in the complete elimination of myocardial gene expression and the absence of cell movements associated with embryonic heart development. This inhibition can be rescued by expression of wild type tinman sequences. These experiments indicate that function of tinman family genes is essential for development of the vertebrate heart. PMID- 9851853 TI - Distinct regulatory elements govern Fgf4 gene expression in the mouse blastocyst, myotomes, and developing limb. AB - Embryonic development requires a complex program of events which are directed by a number of signaling molecules whose expression must be rigorously regulated. We previously showed that expression of Fgf4, which plays an important role in postimplantation development and growth and patterning of the limb, is regulated in EC cells by the synergistic interaction of Sox2 and Oct-3 with the Fgf4 EC cell-specific enhancer. To verify whether this mechanism was also operating in vivo, and to identify new elements controlling Fgf4 gene expression in distinct developmental stages, we have analyzed the expression of LacZ reporter plasmids containing different fragments of the Fgf4 gene in transgenic mouse embryos. Utilizing these transgenic constructs we have been able to recapitulate, for the most part, Fgf4 gene expression during embryonic development. We show here that most of the cis-acting regulatory elements determining Fgf4 embryonic expression are located in conserved regions within the 3' UTR of the gene. The EC cell specific enhancer is required to drive gene expression in the ICM of the blastocyst, and its activity requires the Sox and Oct-proteins binding sites. We were also able to identify specific and distinct enhancer elements that govern postimplantation expression in the somitic myotomes and the limb bud AER. The myotome-specific elements contain binding sites for bHLH myogenic regulatory factors, which appear to be essential for myotome expression. Finally, we present evidence that the very restricted pattern of expression of Fgf4 transcripts in the AER results from the combined action of positive and negative regulatory elements located 3' of the Fgf4 coding sequences. Thus the Fgf4 gene relies on multiple and distinct regulatory elements to achieve stage- and tissue-specific embryonic expression. PMID- 9851854 TI - Specific glycoconjugates are present at the oolemma of the fertilization site in the egg of Discoglossus pictus (Anurans) and bind spermatozoa in an in vitro assay. AB - In the egg of the anuran Discoglossus pictus, the site of fertilization is restricted to the central portion of an animal hemisphere indentation (the dimple). Previous studies showed that the acrosome reaction of D. pictus sperm is triggered in the jelly, and yet sperm arrive at the dimple surface with the plasma membrane at an early stage of vesiculation. Reactivity of the dimple surface with specific lectins suggests that fucose might be utilized as a marker of glycoproteins located at the dimple surface. In this paper, proteins of the egg surface were labeled with the membrane impermeable sulfo-NHS-biotin. Four main bands of 200, 230, 260, and 270 kDa labeled only at the dimple surface, although they were detected in the cortex of the whole egg. The 270-kDa band reacted with Galanthus nivalis agglutinin only in the cortex of the dimple, suggesting that this band is differently glycosylated according to its localization. The alpha-l-fucose-specific lectin Ulex europaeus agglutinin I was utilized both in lectin blotting and in affinity chromatography and cross-reacted with the 200- and 270/260-kDa bands. Furthermore, two polypeptides were obtained by exposure of intact eggs to lysylendoproteinase C. They were also reactive to Ulex europaeus agglutinin I. The 200- and 270/260-kDa bands were eluted from the acrylamide gels and adsorbed to polystyrene beads. An assay for sperm binding to 200-kDa glycoprotein-bound beads was developed. Sperm stuck to the beads before but not after Ca-ionophore treatment. When the beads were coated with the 270/260 kDa glycoproteins, binding occurred after ionophore treatment. In these assays, the 200- and 270/260-kDa glycoproteins competitively inhibited sperm binding to the beads coated with the corresponding glycoprotein. These results indicate that the assayed glycoproteins, located either in the glycocalyx or in the plasma membrane of the fertilization site, are involved in sperm binding. PMID- 9851856 TI - Bottle cells are required for the initiation of primary invagination in the sea urchin embryo. AB - Invagination of epithelial tissue occurs during gastrulation, neurulation, and organogenesis in many organisms. However, the underlying morphogenetic mechanisms of invagination are not understood. To elucidate these mechanisms, we have analyzed the initial invagination of the vegetal plate in the sea urchin embryo, a process termed primary invagination. At the onset of invagination, a ring of cells with highly constricted apices (bottle cells) encircles a group of two to eight round, central cells. To investigate the morphogenetic role of the bottle cells in the process of primary invagination, we have undertaken a series of laser ablation studies in which different proportions of various cell types were ablated and the effects were recorded using 4-D microscopy. Elimination of a 90 degrees-180 degrees arc of bottle cells markedly retards invagination, but only within the ablated region. Ablation of other cell types does not result in a statistically significant effect on primary invagination. These studies indicate that the number and arrangement of the bottle cells are critical factors for proper initiation of invagination. In addition, we have used the perturbing anti hyalin antibody mAb183 to show that cell attachment to the hyaline layer is necessary for bottle cell formation and the initiation of primary invagination. PMID- 9851855 TI - Heart malformations in transgenic mice exhibiting dominant negative inhibition of gap junctional communication in neural crest cells. AB - Transgenic mice were generated expressing an alpha1 connexin/beta-galactosidase fusion protein previously shown to exert dominant negative effects on gap junctional communication. RNase protection analysis and assays for beta galactosidase enzymatic activity showed that the transgene RNA and protein are expressed in the embryo and adult tissues. In situ hybridization analysis revealed that in the embryo, expression was predominantly restricted to neural crest cells and their progenitors in the dorsal neural tube, regions where the endogenous alpha1 connexin gene is also expressed. Dye-coupling analysis indicated that gap junctional communication was inhibited in the cardiac neural crest cells. All of the transgenic lines were homozygote inviable, dying neonatally and exhibiting heart malformations involving the right ventricular outflow tract-the same region affected in the alpha1 connexin knockout mice. As in the knockout mice, the conotruncal heart malformations were accompanied by outflow tract obstruction. Histological analysis showed that this was associated with abnormalities in the differentiation of the conotruncal myocardium. These results suggest that the precise level of gap junctional communication in cardiac neural crest cells is of critical importance in right ventricular outflow tract morphogenesis. Consistent with this possibility is the fact that cardiac crest cells from the alpha1 connexin knockout mice also exhibited a greatly reduced level of gap junctional communication. These studies show the efficacy of a dominant negative approach for manipulating gap junctional communication in the mouse embryo and demonstrate that targeted expression of this fusion protein can be a powerful tool for examining the role of gap junctions in mammalian development. PMID- 9851857 TI - Germ-line regulation of the Caenorhabditis elegans sex-determining gene tra-2. AB - The Caenorhabditis elegans sex-determining gene tra-2 promotes female development of the XX hermaphrodite soma and germ line. We previously showed that a 4.7-kb tra-2 mRNA, which encodes the membrane protein TRA-2A, provides the primary feminizing activity of the tra-2 locus. This paper focuses on the germ-line activity and regulation of tra-2. First, we characterize a 1.8-kb tra-2 mRNA, which is hermaphrodite-specific and germ-line-dependent. This mRNA encodes TRA 2B, a protein identical to a predicted intracellular domain of TRA-2A. We show that the 1.8-kb mRNA is oocyte-specific, suggesting that it is involved in germ line or embryonic sex determination. Second, we identify a tra-2 maternal effect on brood size that may be associated with the 1.8-kb mRNA. Third, we investigate seven dominant tra-2(mx) (for mixed character) mutations that sexually transform hermaphrodites to females by eliminating hermaphrodite spermatogenesis. Each of the tra-2(mx) mutants possesses a nonconserved missense change in a 22-amino-acid region common to both TRA-2A and TRA-2B, called the MX region. We propose that the MX region mediates a posttranslational regulation of tra-2 essential for the onset of hermaphrodite spermatogenesis. Finally, we discuss aspects of tra-2 function and regulation that are specific to the unusual control of cell fate in the hermaphrodite germ line. PMID- 9851858 TI - The cellular mechanism of epithelial rearrangement during morphogenesis of the Caenorhabditis elegans dorsal hypodermis. AB - The mechanism by which epithelial cells rearrange is a process that is central to epithelial morphogenesis, yet remains poorly understood. We have investigated epithelial cell rearrangement in the dorsal hypodermis of the Caenorhabditis elegans embryo, in which two rows of epithelial cells rearrange in a morphogenetic process known as dorsal intercalation. The intercalating cells extend basal protrusions which squeeze between their opposing neighbors beneath their adherens junctions. As the intercalating cells move forward, these protruding tips become broader in the anterior-posterior and dorsoventral dimensions, effectively "plowing through" the adherens junctions and forcing an opening for the remainder of the intercalating cell to insert between the contralateral cells. These cell movements are dependent upon intact cytoarchitecture, since the pharmacological disruption of microtubules or actin filaments blocks cell rearrangement. The cells appear to intercalate independently of immediately adjacent neighboring hypodermal cells because dorsal intercalation is not blocked by the ablation of the progenitors for either half of the lateral hypodermal cells or the posterior half of the dorsal hypodermis. This is the first case in which the protrusive mechanism underlying epithelial cell rearrangement has been characterized, and we propose a model describing how epithelial cells rearrange within the confines of an epithelial monolayer, and discuss the mechanisms that may be guiding these directed cell movements. PMID- 9851859 TI - Activation of the Jak-STAT-signaling pathway in embryonic lens cells. AB - Previous studies showed that lens epithelial cells proliferate rapidly in the embryo and that a lens mitogen, most likely derived from the blood, is present in the anterior chamber of the embryonic eye (Hyatt, G. A., and Beebe, D. C., Development 117, 701-709, 1993). Messenger RNAs for several growth factor receptors have been identified in embryonic lens epithelial cells. We tested several growth factors that are ligands for these receptors for their ability to maintain lens cell proliferation. Embryo serum, PDGF, GM-CSF, and G-CSF maintained lens cell proliferation, but NGF, VEGF, and HGF did not. This and a previous study (Potts, J. D., Harocopos, G. J., and Beebe, D. C., Curr. Eye Res. 12, 759-763, 1993) detected members of the Janus kinase family (Jaks) in the developing lens. Because Jaks are central players in the Jak-STAT-signaling pathway, we identified STAT proteins in the lens and tested whether they were phosphorylated in response to mitogens. STAT1 and STAT3, but not STAT 5 were detected in chicken embryo lens epithelial cells. Only STAT3 was found in terminally differentiated lens fiber cells. STAT1 and STAT3 were phosphorylated in lens cells analyzed immediately after removal from the embryo and when lens epithelial explants were treated with embryo serum, PDGF, or GM-CSF, but not with NGF. Chicken embryo vitreous humor or IGF-1, factors that stimulate lens cell differentiation, but not proliferation, did not cause STAT phosphorylation. When lens epithelial cells were cultured for 4 h in unsupplemented medium, STAT1 and STAT3 declined to nearly undetectable levels. Treatment with PDGF or embryo serum for an additional 15 min restored STAT1 and -3 levels. This recovery was blocked by cycloheximide, but not actinomycin D, suggesting that STAT levels are regulated at the level of translation. STAT levels were maintained in epithelial explants by lens mitogens, but not by factors that stimulated lens fiber differentiation. Both factors that stimulated lens cell proliferation and those that caused fiber differentiation protected cultured lens epithelial cells from apoptosis. These data suggest that the factor(s) responsible for lens cell proliferation in vivo activates the Jak-STAT-signaling pathway. They also indicate that growth factors maintain STAT protein levels in lens epithelial cells by promoting the translation of STAT mRNA, an aspect of STAT regulation that has not been described previously. Signaling by most of the growth factors and cytokines known to activate the Jak-STAT pathway has been disrupted in mice by mutation or targeted deletion. Consideration of the phenotypes of these mice suggests that the factor responsible for lens cell proliferation in vivo may be a growth factor or cytokine that has not yet been described. PMID- 9851862 TI - Peter Thorogood (1947-1998). PMID- 9851860 TI - The TATA binding protein in the sea urchin embryo is maternally derived. AB - The cDNA encoding the TATA binding protein was isolated from 8- to 16-cell and morula-stage embryonic libraries of two distantly related species of sea urchin, Strongylocentrotus purpuratus and Lytechinus variegatus, respectively. The two proteins are 96% identical over both the N- and C-terminal domains, suggesting a conservation of transcriptional processes between the two species. The prevalence of SpTBP transcripts at several developmental time points was determined using the tracer excess titration method, and the corresponding number of TBP protein molecules was determined by quantitative Western blot analysis. Our results indicate that the amount of TBP mRNA and protein per embryo remains relatively constant throughout development. An initial large pool of TBP protein (>10(9)) molecules in the egg becomes diluted as a consequence of cell division and decreases to about 2 x 10(6) molecules per cell by the gastrula stage. We found by in situ RNA hybridization that the oocyte contains a large amount of TBP mRNA which is depleted late in oogenesis so that the eggs and early embryos have extremely low levels of TBP mRNA. We conclude that the oocyte manufactures nearly all of the TBP protein necessary for embryogenesis. PMID- 9851861 TI - The 350-kDa sea urchin egg receptor for sperm is localized in the vitelline layer. AB - Previous studies have established by several methods that the 350-kDa egg receptor for sperm is localized on the plasma membrane-vitelline layer complex of the egg of the sea urchin Strongylocentrotus purpuratus. In addition, it has been found that molecules which are cross-reactive with anti-receptor antibody are present in the cortical granules located at the inner face of the plasma membrane. The objective of this study was to define more precisely the locale of the cell surface receptor. We have found that following fertilization, the immunoreactive receptor initially found on the egg surface moved to the fertilization envelope (FE) and then disappeared in parallel with the loss of sperm binding activity. A cross-linked, high-molecular-weight derivative of soybean trypsin inhibitor (hMW-SBTI) which was unable to pass through the elevating FE blocked the loss of both immunoreactivity and the sperm binding activity of the FE, but did not inhibit the vitelline delaminase activity that has been implicated in FE formation. Western blot analysis following SDS-PAGE of the proteins of the FE isolated in the presence of hMW-SBTI and benzamidine revealed the presence of the 350/320-kDa proteins which cross-reacted with anti receptor antibody. Experiments in which molecules on the surface of unfertilized eggs were labeled with biotin and traced after FE formation revealed that a significant portion of the 350/320-kDa glycoproteins that were incorporated into the FE originated from the cell surface, rather than from the cortical granules. These findings provide strong evidence that in unfertilized eggs the egg receptor for sperm exists as part of the protein complex known as the vitelline layer which serves as a precursor of the FE. Evidence is presented indicating that some of the receptor in the vitelline layer is cryptic and a possible function for this cryptic form of the receptor is proposed. PMID- 9851863 TI - Alterations in both heparan sulfate proteoglycans and mitogenic activity of fibroblast growth factor-2 are triggered by inhibitors of proliferation in normal and breast cancer epithelial cells. AB - Heparan sulfate proteoglycans (HSPG) are involved in the regulation of cellular proliferation, differentiation, and migration. We have studied the effect of three inhibitors of proliferation on 35S incorporation into HSPG of the breast cancer cell lines MCF-7 and MDA-MB-231 and the normal breast epithelial cells (NBEC). Transforming growth factor beta-1 (TGFbeta-1), which inhibits the proliferation of NBEC, but not of MCF-7 and MDA-MB-231, cells induced an increase in 35S incorporation of HSPG in NBEC, but had no effect on cancer cells. Sodium butyrate (NaB), which inhibits NBEC as well as cancer cell proliferation, induced an increase in 35S incorporation into HSPG in all cell types studied. In contrast, retinoic acid had no effect on HSPG of breast epithelial cells. Modification of HSPG induced by TGFbeta-1 or NaB treatments in normal and breast cancer epithelial cells resulted in an increase in 125I-fibroblast growth factor 2 (FGF-2) binding on HSPG. More importantly, NaB pretreatment resulted in an inhibition of the MCF-7 cell responsiveness to FGF-2, even though these cells remained sensitive to growth stimulation induced by serum or epidermal growth factor. These results indicate that changes in HSPG production are a key process involved in the mechanism of breast epithelial cell growth regulation. PMID- 9851864 TI - In vitro culture system for iris-pigmented epithelial cells for molecular analysis of transdifferentiation. AB - Dissociated cells of the iris-pigmented epithelium (IPE) from a 1-day-old chick grew in monolayer culture and stably maintained their differentiated state when cultured with standard culture medium. After replacement of the control medium by EdFPH medium, which is effective in inducing dedifferentiation of retinal pigmented epithelium (RPE) cells, all cells rapidly lost pigment granules, proliferated intensively, and dedifferentiated. By further addition of ascorbic acid, dedifferentiated cells accumulated and formed a large number of lentoids. This system provides a useful opportunity for analyzing cellular and molecular mechanism involved in each step of transdifferentiation. Furthermore, Northern blot data indicates that the up-regulation of pax-6 gene could be an important event during lens regeneration as well as during normal lens development. PMID- 9851865 TI - Regulation of astrocyte morphology by RhoA and lysophosphatidic acid. AB - Astrocytes in the CNS undergo morphological changes and start to proliferate after breakdown of the blood-brain barrier. In culture, proliferating astrocytes have a flat, polygonal shape. When treated with cAMP-raising agents, astrocytes adopt a stellate, process-bearing morphology resembling their in vivo appearance. Stellation is accompanied by loss of actin stress fibers and focal adhesions. Lysophosphatidic acid (LPA), a blood-borne mitogen that signals through its cognate G protein-coupled receptor, stimulates DNA synthesis in astrocytes and causes rapid reversal of cAMP-induced stellation. LPA reversal of stellation is initiated by f-actin reassembly and tyrosine phosphorylation of focal adhesion proteins such as paxillin. Botulinum C3 toxin, which inactivates the Rho GTPase, mimics cAMP-raising agents in inducing stellation, f-actin disassembly, paxillin dephosphorylation, and growth arrest. However, unlike cAMP-induced stellation, C3 induced stellation cannot be reversed by LPA. Conversely, astrocytes expressing activated RhoA fail to undergo cAMP-induced stellation. Thus, RhoA controls astrocyte morphology in that active RhoA directs LPA reversal of stellation, while inactivation of RhoA is sufficient to induce stellation. PMID- 9851866 TI - Proteinase inhibitor 6 (PI-6) expression in human skin: induction of PI-6 and a PI-6/proteinase complex during keratinocyte differentiation. AB - Proteinase inhibitor 6 (PI-6) is a 42-kDa intracellular protein present in epithelial cells and endothelial cells. It is capable of inhibiting a number of serine proteinases, including trypsin and chymotrypsin. In this study we examined PI-6 expression in human skin and its primary cell type, the keratinocyte. By immunohistochemical analysis, PI-6 staining is absent from the basal cells, weak in the spinous layer, and strongest in the granulosa layer of human epidermis. Immunoblotting of cultured primary keratinocytes revealed that PI-6 production increases 24-fold on differentiation. Analysis of an immortalized keratinocyte cell line, HaCat, showed a 5-fold increase in PI-6 mRNA and a 7-fold increase in PI-6 protein upon differentiation, and indirect immunofluorescence revealed that this is due to an increase in the number of differentiated cells expressing high levels of PI-6. Of particular interest is the appearance of a preformed complex between PI-6 and an endogenous serine proteinase in differentiating HaCat cells, which was detected by a monoclonal antibody demonstrated to preferentially recognize PI-6 in complex with a proteinase. This identification of a PI 6/proteinase complex is the first example of a serpin bound to a proteinase in keratinocytes. We postulate that a physiological role of PI-6 is to regulate a serine proteinase associated with keratinocyte differentiation. PMID- 9851867 TI - Identification of a novel mRNA-associated protein in oocytes of Pleurodeles waltl and Xenopus laevis. AB - Amphibian oocytes accumulate a large pool of mRNA molecules for future embryonic development. Due to their association with specific proteins the stored maternal RNAs are translationally repressed. The identification of these RNA-binding proteins and the characterization of their functional domains may contribute to the understanding of the translational repression mechanisms and the subsequent activation processes during early embryogenesis. Here we present the complete Pleurodeles cDNA sequence of a cytoplasmic protein which is present in oocytes, eggs, and very early cleavage stage embryos but undetectable in postcleavage embryo and adult tissues. The predicted molecular mass of the protein is 55 kDa and the apparent molecular mass as determined by SDS-PAGE, 68 kDa. The deduced amino acid sequence reveals proline- and serine-rich domains in the aminoterminal part as well as two RGG boxes which represent characteristic motifs of several RNA-binding proteins. No distinct homologies to the consensus RNA recognition motif were found. The 55-kDa protein was recovered in cytoplasmic ribonucleoprotein (RNP) particles containing poly(A)+ RNA. It was therefore termed RAP55 for mRNA-associated protein of 55 kDa. However, a direct interaction of RAP55 with mRNA could not be demonstrated by UV-crosslinking experiments, indicating that it is bound to mRNP complexes via protein-protein interactions. RAP55 is evolutionarily conserved since antibodies raised against a recombinant Pleurodeles RAP55 fragment recognize the protein from Pleurodeles and Xenopus. The expression pattern and intracellular distribution of RAP55 suggest that it is part of those mRNP particles which are translationally repressed during oogenesis and become activated upon progesterone-induced oocyte maturation. PMID- 9851868 TI - Evidence for different MCM subcomplexes with differential binding to chromatin in Xenopus. AB - MCM proteins are molecular components of the DNA replication licensing system in Xenopus. These proteins comprise a conserved family made up of six distinct members which have been found to associate in large protein complexes. We have used a combination of biochemical and cytological methods to study the association of soluble and chromatin-bound Xenopus MCM proteins during the cell cycle. In interphase, soluble MCM proteins are found organized in a core salt resistant subcomplex that includes MCM subunits which are known to have high affinity for histones. The interphasic complex is modified at mitosis and the subunit composition of the resulting mitotic subcomplexes is distinct, indicating that the stability of the MCM complex is under cell cycle control. Moreover, we provide evidence that the binding of MCM proteins to chromatin may occur in sequential steps involving the loading of distinct MCM subunits. Comparative analysis of the chromatin distribution of MCM2, 3, and 4 shows that the binding of MCM4 is distinct from that of MCM2 and 3. Altogether, these data suggest that licensing of chromatin by MCMs occurs in an ordered fashion involving discrete subcomplexes. PMID- 9851869 TI - The density of a homogeneous population of cells controls resetting of the program for swarmer formation in the unicellular marine microorganism Noctiluca scintillans. AB - Noctiluca scintillans is a luminescent marine dinoflagellate. The life cycle of Noctiluca consists of a vegetative stage and a swarmer stage. The swarmer stage of Noctiluca is initiated by formation of a swarmer-mother cell instead of binary fission of vegetative cells. We studied the formation of swarmers under various conditions and became convinced that the cells have a strict program for the formation of swarmers which starts to operate in every cell after a defined number of cell cleavages. The probability that the program will be executed appeared to be affected by the presence of other cells. In other words, a high density of cells suppressed the expression of the program. Suppression was achieved by resetting the mechanism and was related to the number of cell divisions. Our findings provide one of the simplest examples of a mechanism by which a large population produces individuality in a group of genetically homogeneous organisms. PMID- 9851870 TI - Galectin-3 stimulates cell proliferation. AB - Galectin-3, a beta-galactoside-binding protein, has been shown to be involved in multiple biological processes through interaction with its complementary glycoconjugates. Here we provide the first evidence of galectin-3 as a mitogen. Incubation of quiescent cultures of normal human lung fibroblast IMR-90 cells with recombinant galectin-3 (rgalectin-3) stimulated DNA synthesis as well as cell proliferation in a dose-dependent manner. This mitogenic activity was dependent on the lectin property of galectin-3, as it could be significantly inhibited by lactose, a disaccharide competitive for carbohydrate-binding by galectin-3. Chemical cross-linking and affinity-purification experiments identified binding of rgalectin-3 to cell surface glycoproteins, which were not recognized by antibodies directed against lysosome-associated membrane proteins (LAMPs), putative cellular ligands for galectin-3. Moreover, pulse-chase analysis revealed no secretion of galectin-3 by IMR-90 cells. These results indicate that galectin-3 is a mitogen capable of stimulating fibroblast cell proliferation in a paracrine fashion through interaction with cell surface glycoconjugates different from LAMPs and suggest a possible involvement of galectin-3 in tissue remodeling. PMID- 9851871 TI - Apoptosis-independent alterations in membrane dynamics induced by curcumin. AB - Curcumin is a well-known natural compound with antiinflammatory properties. Its antiproliferative effect and ability to modulate apoptotic response are considered essential in cancer therapy. The physicochemical properties of curcumin suggest membranous localization, which prompted an investigation of the mechanisms of membrane disturbances evoked by curcumin. We chose the erythrocyte as a convenient model for studying membrane effects of curcumin and showed its nonspecific, apoptosis-independent way of action. Curcumin was found to expand the cell membrane, inducing echinocytosis. Changes in cell shape were accompanied by transient exposure of phosphatidylserine. Membrane asymmetry was recovered by the action of aminophospholipid translocase, which remained active in the presence of curcumin. Lipids rearrangements and drug partitioning caused changes of lipid fluidity. Such nonspecific effects of curcumin on cellular membranes would produce artifacts of apoptosis measurement, since several methods are based on membrane changes. PMID- 9851872 TI - Induction of differentiation and apoptosis in human promyelocytic leukemia HL60 cell line by a new type of steroids. AB - Mer-NF8054X is a new type of steroid whose structure has been established as 11 oxo-18, 22-cycloergosta-6, 8(14)-diene-3beta, 5beta, 9beta, 23S-tetraol (an 18, 22-cycloergostane), which has been reported to have antifungal activity against Aspergillus fumigatus. However, other biological activities are unknown. Herein, we reported that Mer-NF8054X inhibited cell growth of HL60 human leukemia cells, when used either singly or in combination with retinoic acid (RA). In addition, Mer-NF8054X alone induced differentiation and apoptosis of HL60 cells. The induction of differentiation of HL60 cells by Mer-NF8054X was synergistic in combination with RA. On the other hand, Emesterone A, an analogue of Mer-NF8054X which is missing a hydroxy residue from the third position, showed much lower activity than Mer-NF8054X on the inhibition of cell growth and the induction of cell differentiation and apoptosis. However, Emesterone B, an analogue of Emesterone A which is missing a hydroxy residue from the fifth position, showed higher activity than Emesterone A but lower activity than Mer-NF8054X when examined for the inhibition of cell growth and the induction of cell differentiation and apoptosis. These results suggested that Mer-NF8054X and its analogs may be a new type of differentiation inducing agent. The hydroxy residue at the third position or fifth position in Mer-NF8054X may be necessary, but not essential, for inhibition of growth and induction of both differentiation and apoptosis of HL60 cells. In addition, Mer-NF8054X enhanced the differentiation of HL60 cells induced by RA. Based on these results, Mer-NF8054X may have utility in the clinic in combination with RA for leukemia patients. PMID- 9851873 TI - Identification of chromosomal bands replicating early in the S phase of normal human fibroblasts. AB - Normal human fibroblasts (NHF1) were released from confluence arrest (G0) and replated in medium containing bromodeoxyuridine (BrdU) and aphidicolin. Despite severe reduction in the rate of DNA synthesis by aphidicolin, cells reentering the cell cycle incorporated BrdU at regions of the human genome that replicated very early in S phase. After removal of aphidicolin and BrdU from the tissue culture medium, cells were collected in mitosis. Q-banding with 4', 6-diamidino-2 phenylindole/actinomycin D was used to identify metaphase chromosomes. A monoclonal anti-BrdU antibody and a fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse antibody were used to identify the BrdU-labeled sites. The criterion for scoring DNA replication sites was the detection of FITC fluorescence at homologous regions of both sister chromatids. Early replicating regions mapped within R-bands, but not all R-bands incorporated BrdU. Chromosomal bands 1p36.1, 8q24.1, 12q13, 15q15, 15q22, and 22q13 were labeled in 53% or more of the copies of these chromosomes in the data set, suggesting that these sites replicated very early in S phase. Chromosomal band 15q22 was the most frequently labeled site (64%), which indicates that it contains some of the earliest replicating sequences in normal human fibroblasts. PMID- 9851874 TI - Characterization of Cbl-Nck and Nck-Pak1 interactions in myeloid FcgammaRII signaling. AB - Fc receptors modulate inflammatory processes, including phagocytosis, serotonin and histamine release, superoxide production, and secretion of cytokines. Aggregation of FcgammaRIIa, the low-affinity receptor for monomeric IgG, activates nonreceptor protein tyrosine kinases such as Lyn, Hck, and Syk, potentially driving the phosphorylation of the downstream adaptor proteins, including Cbl and/or Nck. Previous work from our laboratory using interferon gamma-differentiated U937 (U937IF) myeloid cells investigated mechanisms which regulate Fcgamma receptor-induced assembly of adaptor complexes. Herein we report that FcgammaRII receptor signaling in U937IF and HEL cells involves Cbl and Nck, suggesting that Cbl-Nck interactions may link FcgammaRII to downstream activation of Pak kinase. FcgammaRII crosslinking induced the phosphorylation of Cbl and Nck on tyrosine. The alphaCbl immunoprecipitations revealed constitutive binding of Nck and Grb2 to Cbl and FcgammaRII-inducible binding of CrkL to Cbl. The interactions of Cbl with Nck and CrkL were phosphorylation dependent since dephosphorylation of cellular proteins with potato acid phosphatase abrogated binding. GST-Nck fusion protein pulldown experiments show that Cbl and Pak1 bind to the second SH3 domain of Nck. A specific Src inhibitor, PP1, was shown to completely abrogate the FcgammaR-induced superoxide response, correlating with a decrease in Cbl and Nck tyrosine phosphorylation. Our results provide the first evidence that Src is required for FcgammaR activation of the respiratory burst in myeloid cells and suggest that Cbl-Nck, Cbl-Pak1, and Nck-Pak1 interactions may regulate this response. PMID- 9851875 TI - Expression of CCAAT/enhancer binding proteins during porcine preadipocyte differentiation. AB - The expression of three CCAAT/enhancer-binding proteins (C/EBPs) was examined with immunocytochemistry and Western blot analysis during preadipocyte differentiation in porcine stromal vascular (S-V) cell cultures. Regardless of treatment and time in culture, immunoreactivity for all three C/EBP isoforms was restricted to cell nuclei. At day 1, 50 +/- 6% of S-V cells were C/EBPdelta positive, whereas 13 +/- 3 and 11.7 +/- 3% of S-V cells were AD-3 and C/EBPalpha positive, respectively. After 3 days of seeding in fetal bovine serum (FBS) and dexamethasone (DEX), C/EBPdelta; AD-3, and C/EBPalpha-positive cells increased to 67 +/- 5, 42 +/- 4, and 32 +/- 3%, respectively. Double staining clearly showed that most of the C/EBPalpha reactive cells had not accumulated appreciable lipid after 3 days of FBS + DEX. Following 3 days of insulin treatment, the percentage of C/EBPdelta cells was 50 +/- 6, whereas the percentage of AD-3- and C/EBPalpha positive cells was 41 +/- 4 and 31 +/- 3, respectively. After insulin treatment all fat cells were AD-3, C/EBPalpha, and C/EBPdelta positive. Double staining demonstrated that fat cells were C/EBPdelta reactive throughout the culture period. Western blotting showed changes in C/EBP isoform expression that were consistent with the immunocytochemical results. We conclude that C/EBPalpha is a terminal differentiation marker which is expressed later than AD-3 but further studies are needed to determine the relationship between C/EBPdelta and adipogenesis in porcine S-V cultures. PMID- 9851876 TI - Blockade of tumor cell transforming growth factor-betas enhances cell cycle progression and sensitizes human breast carcinoma cells to cytotoxic chemotherapy. AB - We have examined the effect of neutralizing TGF-beta antibodies on cisplatin mediated cytotoxicity against MDA-231 human breast tumor cell spheroids. These tridimensional in vitro systems have been shown to recapitulate the drug sensitivity pattern of tumor cells in vivo. MDA-231 tumor cell spheroids exhibit higher protein levels of the cyclin-dependent kinase (Cdk) inhibitors p21 and p27 and >10-fold lower Cdk2 activity compared to adherent cell monolayers, as well as pRb hypophosphorylation, a predominant G1 population, and a cisplatin 1-h IC50 of approximately 100 microM. Treatment of MDA-231 cells in monolayer with cisplatin for 1 h, subsequently grown as spheroids, increased steady-state TGF-beta1 mRNA levels, secretion of active TGF-beta, cellular Cdk2 activity, pRb phosphorylation, and p21 protein levels, while downregulating p27. Accumulation of cells in G2M and progression into S were noted 48 h after treatment with 100 microM cisplatin. We tested whether drug-induced upregulation of TGF-beta1 and p21, perhaps by preventing cell cycle progression, were protective mechanisms against drug-mediated toxicity by using neutralizing anti-TGF-beta antibodies. Anti-TGF-beta antibodies diminished the induction of p21, enhanced the activation of Cdk2, and facilitated progression into S and G2M following cisplatin treatment. This resulted in a >twofold enhancement of drug-induced DNA fragmentation and a shift in the cisplatin 1-h IC50 from 100 to <10 microM. These data suggest that tumor cell TGF-beta1 may protect from DNA damage and that postchemotherapy administration of TGF-beta inhibitors may facilitate progression beyond G1/S, potentially increasing the efficacy of cytotoxic chemotherapy. PMID- 9851877 TI - Assessment of rapid morphological changes associated with elevated cAMP levels in human orbital fibroblasts. AB - Orbital fibroblasts exhibit a phenotype distinct from that of other types of fibroblasts. Addition of prostaglandin E2 (PGE2) to culture medium elicits a dramatic change in orbital fibroblast morphology. That response is mediated through the generation of cAMP. Orbital fibroblasts can generate high levels of PGE2 through induction by proinflammatory cytokines of prostaglandin endoperoxide H synthase-2 (PGHS-2). Here we compare the influence on fibroblast morphology of exogenous PGE2, forskolin, and 8-br-cAMP to that mediated through PGHS-2 induction by a lymphocyte-derived cytokine. Within a few hours, orbital fibroblasts treated with any of these test compounds appear under phase-contrast microscopy to exhibit a stellate morphology. When these changes were assessed quantitatively by electric cell-substrate impedance sensing (ECIS), it became evident that 8-br-cAMP, forskolin, and PGE2 initiated shape changes within 30 min of addition to the culture medium, while effects of the cytokine were first evident after approximately 3.5 h. Dermal fibroblasts failed to respond to any of these compounds with regard to changes in cellular morphology. Analysis of micromotion, manifested as small impedance fluctuations, revealed that orbital fibroblasts treated with 8-br-cAMP exhibit less motion than did untreated cells. These results suggest that orbital fibroblast shape can be altered by several compounds known to alter intracellular cAMP levels. They demonstrate the utility of ECIS in the assessment of very rapid and dynamic cellular events associated with changes in cell morphology. PMID- 9851878 TI - Role of RAS2 in recovery from chronic stress: effect on yeast life span. AB - The replicative life span of Saccharomyces cerevisiae was previously shown to be modulated by the homologous signal transducers Ras1p and Ras2p in a reciprocal manner. We have used thermal stress as a life span modulator in order to uncover functional differences between the RAS genes that may contribute to their divergent effects on life span. Chronic exposure of cells throughout life to recurring heat shocks at sublethal temperatures decreased their replicative life span. ras2 mutants, however, suffered the largest decrease compared to wild-type and ras1 mutant cells. The decrease was correlated with a substantial delay in resumption of budding upon recovery from these heat shocks, indicating an impaired renewal of cell cycling. Detailed analysis of gene expression showed that, during recovery, ras2 mutants were selectively impaired in down-regulation of stress-responsive genes and up-regulation of growth-promoting genes. Our results suggest that one of the functions of RAS2 in maintaining life span, for which RAS1 does not substitute, is to ensure renewal of growth and cell division after bouts of stress that cells encounter during their life. This activity of RAS2 is effected by the cyclic AMP pathway. Overexpression of RAS2, but not RAS2(ser42) which is deficient in the activation of adenylate cyclase, completely reversed the effect of chronic stress on life span. Thus, RAS2 is limiting for longevity in the face of chronic stress. Since RAS2 is known to down-regulate stress responses, this demonstrates that for longevity the ability to recover from stress is at least as important as the ability to mount a stress response. PMID- 9851879 TI - Heat stress-induced life span extension in yeast. AB - The yeast Saccharomyces cerevisiae has a limited life span that can be measured by the number of times individual cells divide. Several genetic manipulations have been shown to prolong the yeast life span. However, environmental effects that extend longevity have been largely ignored. We have found that mild, nonlethal heat stress extended yeast life span when it was administered transiently early in life. The increased longevity was due to a reduction in the mortality rate that persisted over many cell divisions (generations) but was not permanent. The genes RAS1 and RAS2 were necessary to observe this effect of heat stress. The RAS2 gene is consistently required for maintenance of life span when heat stress is chronic or in its extension when heat stress is transient or absent altogether. RAS1, on the other hand, appears to have a role in signaling life extension induced by transient, mild heat stress, which is distinct from its life-span-curtailing effect in the absence of stress and its lack of involvement in the response to chronic heat stress. This distinction between the RAS genes may be partially related to their different effects on growth-promoting genes and stress-responsive genes. The ras2 mutation clearly hindered resumption of growth and recovery from stress, while the ras1 mutation did not. The HSP104 gene, which is largely responsible for induced thermotolerance in yeast, was necessary for life extension induced by transient heat stress. An interaction between mitochondrial petite mutations and heat stress was found, suggesting that mitochondria may be necessary for life extension by transient heat stress. The results raise the possibility that the RAS genes and mitochondria may play a role in the epigenetic inheritance of reduced mortality rate afforded by transient, mild heat stress. PMID- 9851880 TI - Activation of cysteine proteases in cowpea plants during the hypersensitive response--a form of programmed cell death. AB - There is increasing evidence that the hypersensitive response during plant pathogen interactions is a form of programmed cell death. In an attempt to understand the biochemical nature of this form of programmed cell death in the cowpea-cowpea rust fungus system, proteolytic activity in extracts of fungus infected and uninfected cowpea plants was investigated, using exogenously added poly(ADP-ribose) polymerase as a marker. Unlike the proteolytic cleavage pattern of endogenous poly(ADP-ribose) polymerase in apoptotic animal cells, exogenously added poly(ADP-ribose) polymerase in extracts of fungus-infected plants was proteolytically cleaved into fragments of molecular masses 77, 52, 47, and 45 kDa. In vitro and in vivo protease inhibitor experiments revealed the activation of cysteine proteases, and possibly a regulatory role, during the hypersensitive response. PMID- 9851881 TI - Cumulative subject index for volumes 238-245 PMID- 9851882 TI - A human succinate-ubiquinone oxidoreductase CII-3 subunit gene ending in a polymorphic dinucleotide repeat is located within the sulfonylurea receptor (SUR) gene. AB - We report the cloning of two variant genes encoding the CII-3 subunit of succinate-ubiquinone oxidoreductase complex II. One gene is located within intron 10 of the human sulfonylurea receptor gene. The 3' boundary of this gene ends in a polymorphic dinucleotide repeat. The second gene CII-3b is expressed at a low level and contains a 102-bp internal deletion compared to CII-3 cDNA. These genes should prove valuable in the characterization of Complex II disorders. PMID- 9851883 TI - Regulation of uropathogenic Escherichia coli adhesin expression by DNA methylation. AB - Pap pili play an important role in the pathogenesis of upper urinary tract infections by enabling uropathogenic Escherichia coli to adhere to host epithelial cells. Pap pili are coded for by the pyelonephritis-associated pili (pap) operon, which consists of 11 genes required for the expression and assembly of Pap pili. Expression of Pap pili is regulated by a phase variation mechanism in which the pili expression state of the bacterial population is skewed between phase-on (expression positive) and phase-off (expression negative) states. Pap phase variation is controlled by the cooperative binding of leucine-responsive regulatory protein (Lrp) to two sets of Lrp binding sites in the upstream regulatory region of the pap operon. A single GATC sequence, which is the target site for deoxyadenosine methylase (Dam), is centrally located within each Lrp binding region. Dam plays a critical role in the expression of Pap pili via the formation of DNA methylation patterns. Methylation of GATC-I reduced the affinity of Lrp for pap DNA by about twofold. Conversely, Lrp specifically blocked methylation of pap GATC-I in vitro. These data support the hypothesis that Lrp and Dam compete for binding to GATC-I, and are consistent with previous results indicating that methylation of GATC-I is important for stability of the phase-off state. PMID- 9851884 TI - Amyloidosis in familial mediterranean fever is associated with a specific ancestral haplotype in the MEFV locus. AB - Familial Mediterranean fever (FMF) is a recessive disease characterized by recurrent attacks of inflammation of serosal membranes, and the gene responsible, MEFV, has been recently identified. Amyloidosis is considered to be the most severe complication. Since colchicine is effective in preventing FMF amyloidosis and since this process can develop even prior to the FMF symptoms, lifelong colchicine treatment is recommended for all FMF patients. Identification of the factor which determines amyloidosis will allow treatment to be directed only to those at risk. In order to investigate the association between amyloidosis and MEFV haplotypes, we studied 56 families from three ethnic groups. We compared the haplotypes of FMF patients with and without amyloidosis in each ethnic group separately and identified 14 different MEFV core haplotypes. A significant association (P < 0.004) was found between amyloidosis and a specific core haplotype, 153bp:104bp at markers D16S3370 and D16S2617, respectively. Amyloidosis was present in 20 out of 70 homozygotes for this haplotype and in 6 out of 35 compound heterozygotes for this and other core haplotypes. None of the patients who did not carry this allele had amyloidosis. There was no association between the various haplotypes and severity of the FMF symptoms, age of onset, or age at commencement of colchicine. Further investigation of the MEFV haplotypes in additional patients is recommended as such an association may save many mildly affected or asymptomatic patients with non-amyloidotic genotypes from receiving unnecessary lifelong colchicine treatment. PMID- 9851885 TI - Modification of development by the CFTR gene in utero. AB - The in utero infection of rats at 16-17 days gestation with a recombinant adenovirus carrying the human cystic fibrosis transmembrane conductance regulator (cftr) gene resulted in altered lung development and morphology. These structural alterations prompted an evaluation of concurrent functional changes in the cftr treated lung. CFTR protein could be detected in treated lungs for up to 30 days postinfection, although it was not detected in the intestines at this time. Increased levels of secreted glycoconjugates and lipids were found in lungs treated in utero with human cftr and large vacuoles containing glycoconjugates were detected within cells of the intestines. The scope and durability of these changes suggested that in utero cftr treatment influenced the activity of secretory cells in the developing lung. Altered secretory products in the lungs of cystic fibrosis patients are thought to be associated with increased susceptibility to Pseudomonas aeruginosa infection. We challenged 3-month-old rats (treated in utero with the human cftr gene) with a lethal, intratrachial dose of this bacteria. Rats treated with cftr exhibited enhanced resistance to Pseudomonas infection when compared to controls. These animals displayed little or no associated inflammatory response. No evidence of the adenovirus transgene was detectable at the time of P. aeruginosa inoculation, indicating that continuous ectopic expression of hcftr was not required for enhanced protection. These data demonstrate that in utero, cftr expression influenced the development and function of cells involved in the primary host defense against bacterial infection in the lung. PMID- 9851886 TI - Insulin stimulates binding of phosphofructokinase to cytoskeleton and increases glucose 1,6-bisphosphate levels in NIH-3T3 fibroblasts, which is prevented by calmodulin antagonists. AB - We report here a novel mechanism of insulin action in cultures of NIH-3T3 fibroblasts. Our experiments revealed that in these cells, insulin induced a rapid and transient increase in cytoskeleton-bound phosphofructokinase (EC 2.7.1.11), the rate-limiting enzyme in glycolysis, with a corresponding decrease in soluble (cytosolic) activity. Insulin also induced a slower increase in the levels of glucose 1,6-bisphosphate, the potent activator of cytosolic glycolysis. Both the rapid and the slower stimulatory actions of insulin were prevented by treatment with structurally different calmodulin antagonists, which strongly suggest that calmodulin is involved in these effects of insulin. The present and our previous experiments in muscle suggest that rapid, Ca2+-calmodulin-mediated increase in the binding of glycolytic enzymes to cytoskeleton, as well as the slower increase in glucose 1,6-bisphosphate, may be a general mechanism, in different cells, in signal transduction of insulin. PMID- 9851887 TI - Conductive choline transport by alveolar epithelial plasma membrane vesicles. AB - Choline is an important substrate in alveolar epithelia for both surfactant production and cellular maintenance. The underlying mechanisms of uptake and sites of membrane transport remain uncertain. To test the hypothesis that choline transport occurs at the basolateral side of alveolar epithelia by both Na+ independent and -dependent mechanisms, plasma membrane vesicles were prepared from the apical and basolateral membranes of mature porcine type II pneumocytes. Choline+ transport was assayed by uptake of [3H]choline+ by enriched apical or basolateral vesicles. In the presence of imposed, inside-negative charge gradients, basolateral vesicles exhibited early overshoot of [3H]choline+ uptake unaffected by the presence or absence of external Na+ (541 +/- 53 vs 564 +/- 79 pmol/mg protein (NS)). High sensitivity to hemicholinium-3 was observed in the presence or absence of Na+. In the absence of inside-negative charge gradients, uptake was reduced 12-fold in the presence or absence of Na+, and external choline+ induced internal alkalization of acidified basolateral vesicles. Accumulative [3H]choline+ uptakes by apical vesicles in the presence or absence of inside-negative charge gradients and Na+ were insignificant. We conclude that predominant choline+ uptake by type II pneumocytes occurs at the basolateral membrane by Na+-independent, electrogenic choline+ conductance. The presence of electroneutral choline+/H+ exchange is suggested. PMID- 9851888 TI - Phosphodiesterase activity in intrapulmonary arteries and veins of perinatal lambs. AB - The transition from fetal to newborn life is marked by a reduction in pulmonary vascular tone mediated by the intracellular second messengers, cGMP and cAMP. We have compared the rates of phosphodiesterase (PDE)-catalyzed hydrolysis of cGMP and cAMP in intrapulmonary vessels of fetal (146 +/- 2 days gestation) and newborn (3-7-day-old) lambs, each n = 6. Lung vessels of second to sixth generations were dissected and cytosol was prepared by differential centrifugation. PDE activity in cytosol was determined by radiometric assay of the hydrolysis of exogenous nucleotides at 30 degrees C for 10 min. Rates of hydrolysis (pmol/min/mg protein) of cGMP were 225 +/- 38 in fetal arteries and different from 151 +/- 7 in veins. In newborn vessels, the rates were 155 +/- 49 and 63 +/- 13 in arteries and veins, respectively. Rates of cAMP hydrolysis by the fetus were 80 +/- 11 in arteries and 45 +/- 16 veins. In newborn lambs the rates were 69 +/- 10 in arteries and different from 18 +/- 4 in veins. Inhibition of PDE activity by zaprinast, a cGMP-specific PDE inhibitor, and rolipram, a cAMP specific PDE inhibitor, was more in veins of fetal and newborn lambs. Our data show that rates of hydrolysis of the cyclic nucleotides were faster in fetal vessels than in the newborn. We speculate that this would result in a greater accumulation of the cyclic nucleotides in newborn vessels, particularly the veins, and therefore endow the veins with less vascular tone. PMID- 9851889 TI - Plasma glutathione peroxidase and its relationship to renal proximal tubule function. AB - Selenium-dependent extracellular glutathione peroxidase (E-GPx) is found in plasma and other extracellular fluids. Previous studies have indicated that patients with chronic renal failure on dialysis have low plasma GPx activity. In this study, dialysis patients had approximately 40% of control plasma GPx activity, while anephric individuals had lowest plasma GPx activities ranging from 2 to 22% of control. The residual plasma GPx activity in anephric individuals could be completely precipitated by anti-E-GPx antibodies, indicating that all plasma GPx activity can be attributed to E-GPx in both normal and anephric individuals. Plasma GPx activity rises rapidly following kidney transplantation, often reaching normal values within 10 days. The plasma GPx activity in some transplanted patients rises to levels higher than the normal range, followed by a return to the normal range. Since E-GPx in the kidney is primarily synthesized in the proximal tubules, we investigated whether nephrotoxic agents known to disrupt proximal tubule function also affected plasma GPx activity. The beta-lactam antibiotic cephaloglycin rapidly caused a decrease in plasma GPx activity in rabbits. In addition, the chemotherapeutic agent ifosfamide caused a decrease in plasma GPx activity in pediatric osteosarcoma patients. Fanconi syndrome associated with either ifosfamide therapy or valproic acid therapy also caused a decrease in plasma GPx activity. Thus plasma GPx activity is related to kidney function and is decreased in certain situations where nephrotoxic drugs are administered. Monitoring plasma GPx activity may have predictive value in evaluating the function of transplanted kidneys or in predicting those patients particularly at risk of nephrotoxic injury associated with certain medications. PMID- 9851890 TI - Studies on human porin XVIII: the multicompartment effector ruthenium red reduces the voltage dependence of human VDAC in planar lipid bilayers. AB - The initial data on the effect of ruthenium red on mature human type-1 VDAC are presented. Highly enriched human type-1 porin in planar lipid bilayers shows lowered voltage-dependence whenever a commercially available ruthenium red preparation is applied. The hexavalent polycationic dye ruthenium red affects different functions in varying cell compartments. Concerning the plasma membrane of cells the actual data, together with our former measurements on the interaction of VDAC and the polycationic synthetic polyamine Compound 48/80, refer to a second VDAC opener, which is relevant for studies on the stimulation of exocytotic processes of different cell types. PMID- 9851892 TI - Three new mutations in a gene causing Hermansky-Pudlak syndrome. PMID- 9851891 TI - At least six different mutations in HEXA gene cause Tay-Sachs disease among the Turkish population. AB - Twenty-five Turkish infants with Tay-Sachs disease (TSD) have been diagnosed in the past 8 years. All are from consanguineous, nonrelated families. The present study deals with the molecular basis of six Turkish TSD patients from five unrelated families in which the parents were first cousins. The five mutations identified in this study were INS-5 G-->A, R393X, R137X, 12-bp deletion in exon 10, and G454D. The first three were reported in earlier studies, two in Turkish TSD infants and one in a French TSD infant. PMID- 9851894 TI - alpha2-macroglobulin and C1-inactivator are plasma inhibitors of human glandular kallikrein. AB - Human glandular kallikrein (hK2) is a possible new marker for prostate cancer that is homologous to prostate specific antigen. Purified hK2 added to serum or plasma reacted with endogenous protease inhibitors to form complexes of >350, 135, and 80 kDa, and some hK2 remained free, as judged by immunoblotting. The former two complexes could be removed by specific antibodies to alpha2 macroglobulin and to C1- inactivator, respectively, and they comigrated on SDS PAGE with complexes formed between hK2 and purified alpha2-macroglobulin or C1 inactivator. hK2 complexes of 80 kDa could not be completely removed with any anti-serpin antibody used. Thus, these may consist of more than one type of hK2 complex. In contrast, essentially all hK2 complexes were removed from seminal plasma by antibody to protein C inhibitor, demonstrating that protein C inhibitor is the only significant inhibitor of hK2 in semen. hK2 reacted more rapidly with alpha2-macroglobulin than with any other inhibitor in plasma or serum. Divalent metal ions and heparin did not appreciably affect the rate of formation of any of the hK2 complexes in serum or plasma or with purified alpha2-macroglobulin or C1 inactivator. Measurement of one or more of the hK2 forms identified here may have diagnostic or prognostic potential for prostate cancer. PMID- 9851893 TI - Expression, purification, and characterization of a recombinant erythroid specific hexokinase isozyme. AB - Hexokinase type I (HK I; ATP: D-hexose 6-phosphotransferase, EC 2.7.1.1), the predominant glucose-phosphorylating enzyme in red blood cells, exists in human erythrocytes in multiple molecular forms that differ in isoelectric point and are separable by ion-exchange chromatography. The major forms, designated HK Ia, Ib and Ic, have similar kinetic properties but are characterized by different age dependent decay and different intracellular distribution in reticulocytes. HK Ib, which elutes between HK I and HK II in the DEAE ion-exchange chromatography, appears to be unique to RBCs and different from any other hexokinase isozyme previously described. Indeed, Murakami and Piomelli recently reported the presence of a specific HK isozyme (named HKr) expressed in K562 cells and in human reticulocytes and, moreover, the resolution of the human HK I gene structure provided the direct evidence of an erythroid-specific exon 1. To further investigate the microheterogeneity of HK I in human RBCs we established a prokaryotic expression system for the HKr isozyme, using the pET plasmid, inducible with IPTG. The recombinant HKr, expressed in bacterial cells as a catalytically active enzyme, was purified to homogeneity by a combination of DEAE ionexchange chromatography followed by hydrophobic interaction chromatography and dye-ligand affinity chromatography. The kinetic and chromatographic properties of the homogeneous recombinant HKr suggest that this erythroid-specific HK isozyme in fact corresponds to the HK isoform previously described in human RBCs and referred to as HK Ib. PMID- 9851895 TI - A novel complex allele and two new point mutations in type 2 (acute neuronopathic) Gaucher disease. AB - Gaucher disease, the most prevalent inherited sphingolipidosis, is characterized by lipid laden histiocytes in the spleen, liver and bone marrow sinusoids of affected individuals. It results from deleterious mutations in the functional gene of glucocerebrosidase (acid beta-glucosidase, EC. 3.2.1.45) and is transmitted as an autosomal recessive trait. Three clinical forms of Gaucher disease have been described: Type 1 non-neuronopathic, type 2 acute neuronopathic, and type 3 subacute neuronopathic. In this report, we describe the identification and characterization of three novel mutations from two patients who died with type 2 Gaucher disease. Two heterozygous missense point mutations, one at cDNA nucleotide 238A (E41L) and the other at cDNA nucleotide 508T (R131C) were identified, both in the context of a cDNA nucleotide 1448C (L444P) mutation in the second allele. One of these L444P mutations was identified as a novel complex allele resulting from a crossover involving the glucocerebrosidase functional gene and pseudogene beginning between genomic nucleotides 5689 and 5723 and extending through the rest of the coding sequence. Based on the recent identification and sequence analysis of the metaxin gene and pseudogene contiguous with the glucocerebrosidase pseudogene and functional gene respectively, we have developed a PCR-based method for the analysis of the origin and extent of this recombination. PMID- 9851896 TI - Hemochromatosis in Ireland and HFE. AB - Sixty patients diagnosed with hereditary hemochromatosis with grade 3 or 4 hepatic iron overload and 18 patients diagnosed with hereditary hemochromatosis who had less than grade 3 hepatic iron overload were examined for the HFE gene mutations, 845A (C282Y) and 187G (H63D). Control samples were obtained from 109 randomly selected individuals. Fifty-six of 60 unrelated hereditary hemochromatosis patients (93%) with grade 3 or 4 hepatic iron deposition were homozygous for the C282Y mutation. Fourteen of the 18 hereditary hemochromatosis patients with <3+ iron deposition (76%) were homozygous for the C282Y mutation. Three of 8 patients who were heterozygous for the C282Y mutation were also heterozygous for the H63D mutation. Thirty-one of 109 control individuals were heterozygous for the C282Y mutation and 27 were heterozygous for the H63D mutation. Our finding that 93% of hereditary hemochromatosis patients who fulfil standard diagnostic criteria are homozygous for the C282Y mutation provides clear evidence that this mutation is strongly associated with hereditary hemochromatosis. The allele frequency of 14% for the C282Y mutation in our control population is the highest reported and supports the hypothesis of a Celtic origin for the hereditary hemochromatosis gene. PMID- 9851897 TI - Celtic origin of the C282Y mutation of hemochromatosis. AB - The C282Y mutation in the HFE gene is the main mutation causing hemochromatosis, and C282Y frequencies have been reported for various European populations. The aim of this review is to compile the Y allele frequencies of the C282Y mutation for twenty European populations. The most elevated value (6.88%) is observed in residual Celtic populations in UK and France, in accordance to the hypothesis of Simon et al. concerning a Celtic origin of the hereditary hemochromatosis mutation. PMID- 9851899 TI - Colour vision of the foraging swallowtail butterfly papilio xuthus AB - This paper demonstrates that foraging summer-form females of the Japanese yellow swallowtail butterfly Papilio xuthus have colour vision. The butterflies were trained to feed on sucrose solution placed on a disk of a particular colour in a cage set in the laboratory. After a few such training runs, a butterfly was presented with the training colour randomly positioned within an array of disks of other colours, but with no sucrose solution. The results indicate that the butterflies learn rapidly to select the training colour reliably among different colours. The training colour was also correctly selected when it was covered with neutral density filters to reduce its brightness, or even when the colour was presented together with disks of a variety of shades of grey. These results demonstrate convincingly, for the first time, that a butterfly has true colour vision. PMID- 9851900 TI - Patterned activation of unpaired median neurons during fictive crawling in manduca sexta larvae AB - The unpaired median neurons are common to the segmental ganglia of many insects. Although some of the functional consequences of their activation, among them the release of octopamine to modulate muscle contraction, have been described, less is understood about how and when these neurons are recruited during movement. The present study demonstrates that peripherally projecting unpaired median neurons in the abdominal and thoracic ganglia of the larval tobacco hornworm Manduca sexta are recruited rhythmically during the fictive crawling motor activity that is produced by the isolated central nervous system in response to pilocarpine. Regardless of the muscles to which they project, the efferent unpaired median neurons in all segmental ganglia are depolarized together during the phase of the crawling cycle when the thoracic leg levator motoneurons are active. During fictive crawling, therefore, the unpaired median neurons are not necessarily active in synchrony with the muscles to which they project. The rhythmical synaptic drive of the efferent unpaired median neurons is derived, at least in part, from a source within the subesophageal ganglion, even when the motor pattern is evoked by exposing only the more posterior ganglia to pilocarpine. In pairwise intracellular recordings from unpaired median neurons in different ganglia, prominent excitatory postsynaptic potentials, which occur with an anterior-to-posterior delay in both neurons, are seen to underlie the rhythmic depolarizations. One model consistent with these findings is that one or more neurons within the subesophageal ganglion, which project posteriorly to the segmental ganglia and ordinarily provide unpatterned synaptic inputs to all efferent unpaired median neurons, become rhythmically active during fictive crawling in response to ascending information from the segmental pattern generating network. PMID- 9851901 TI - The synaptic junctions of LE and RF cluster sensory neurones of Aplysia californica are differentially modulated by serotonin. AB - The monosynaptic component of withdrawal reflexes in Aplysia californica, from sensory neurones to motor neurones, is a critical site of the synaptic modulation occurring during short-term and long-term behavioural changes. There are four clusters of sensory neurones (LE, rLE, RE, RF) innervating the receptive field for the gill and siphon withdrawal reflex. The receptive fields of these cells are located on the siphon, the mantle, the branchial cavity and the gill itself. In most studies, the synapses made by the sensory neurones of the LE cluster of the abdominal ganglion or the VC cluster of the pleural ganglion have been investigated and shown to be facilitated by the neuromodulator serotonin (5 hydroxytryptamine, 5-HT). In this report, we have examined the effect of 5-HT on the synaptic junctions of the RF cluster neurones. The duration of action potentials in these cells, unlike those of the other clusters, is barely affected by serotonin. We found that while the LE synapses are facilitated by 5-HT (10 micromol l-1), the RF synapses are not. In fact, the RF-L14 connections are actually depressed by 5-HT; this effect is not due to shunting in the postsynaptic neurone. The RF-L7 connections are also depressed by 5-HT, although the effect is smaller. The RF-L14 connections are blocked by the non-NMDA receptor agonist CNQX (100 micromol l-1), suggesting that the transmitter and the postsynaptic receptors involved are similar to those present on the LE or VC cluster cells. The absence of serotonin-induced facilitation of the RF cluster cells may provide the animal with a means of reducing the nonspecific effects of aversive sensitization and therefore of allowing a greater specificity and more flexibility in plastic behavioural changes. PMID- 9851902 TI - Directional hearing in grasshoppers: neurophysiological testing of a bioacoustic model AB - A recently proposed biophysical model for directional hearing in grasshoppers was tested using complex stimulus situations, with two loudspeakers, one on either side of the animal, synchronously emitting sinusoids with defined phase and amplitude relationships. Hearing responses were determined from whole nerve recordings and compared with the predictions of the model. In Schistocerca gregaria, there were only minor differences between the predictions of the model and measurements and, by reducing the value of the gain of the internal sound path measured previously, a close agreement was achieved between model and measured hearing responses. In Chorthippus biguttulus, larger discrepancies between model calculations using the values measured previously and neuronal response functions were found in both shape and amplitude. A better fit between measurements and model predictions was achieved by increasing the values of the internal delay over those measured previously. The measurements presented here indicate high inter-individual variability of the parameters of the internal pathway, with a range of 60 degrees for the internal phase delay. Calculating the directional characteristics using this range of values for the internal delay indicated that sufficient directional information was available down to 5 kHz. Increasing the value of the internal delay over that measured in an earlier study therefore provides an explanation for the discrepancy between the poor directional information attributed to C. biguttulus in that study and the excellent lateralization ability of males of this species at 5 kHz. PMID- 9851903 TI - Elastic energy storage and release in white muscle from dogfish scyliorhinus canicula AB - The production of work by the contractile component (CC) and the storage and release of work in the elastic structures that act in series (the series elastic component, SEC) with the contractile component were measured using white muscle fibres from the dogfish Scyliorhinus canicula. Heat production was also measured because the sum of work and heat is equivalent to the energy cost of the contraction (ATP used). These energy fluxes were evaluated in contractions with constant-velocity shortening either during stimulation or during relaxation. The muscle preparation was tetanized for 0.6 s and shortened by 1 mm (approximately 15 % of L0) at 3.5 or 7.0 mm s-1 (approximately 15 or 30 % of V0), where L0 is the muscle length at which isometric force is greatest and V0 is the maximum velocity of shortening. In separate experiments, the stiffness of the SEC was characterized from measurements of force responses to step changes in the length of contracting muscle. Using the values of SEC stiffness, we evaluated separately the work and heat associated with the CC and with the SEC. The major findings were (1) that work stored in the SEC could be completely recovered as external work when shortening occurred during relaxation (none of the stored work being degraded into heat) and (2) that, when shortening occurred progressively later during the contraction, the total energy cost of the contraction declined towards that of an isometric contraction. PMID- 9851904 TI - Effects of incline and speed on the three-dimensional hindlimb kinematics of a generalized iguanian lizard (Dipsosaurus dorsalis) AB - Lizards commonly move on steep inclines in nature, but no previous studies have investigated whether the kinematics of the limbs of lizards differ on inclined surfaces compared with level surfaces. Therefore, we examined how the kinematics of the hindlimb were affected by both incline (downhill 30 degrees, level and uphill 30 degrees) and different speeds of steady locomotion (50-250 cm s-1) in the morphologically generalized iguanian lizard Dipsosaurus dorsalis. On the uphill surface, the strides of lizards were shorter and quicker than those at a similar speed on the level and downhill surfaces. A multivariate analysis revealed that the kinematics of locomotion on all three inclines were distinct, but several kinematic features of locomotion on the downhill surface were especially unique. For example, downhill locomotion had the lowest angular excursions of femur rotation, and the knee and ankle were flexed more at footfall which contributed to a very low hip height. For D. dorsalis, changes in knee and ankle angles on the uphill surface were similar to those described previously for mammals moving up inclines, despite fundamental differences in limb posture between most mammals and lizards. Several features of the kinematics of D. dorsalis suggest that a sprawling limb enhances the ability to move on inclines. PMID- 9851905 TI - Cholecystokinin affects gastric emptying and stomach motility in the rainbow trout Oncorhynchus mykiss. AB - In this study, we describe new methods for recording gastric emptying and in vivo measurements of intragastric pressure in fish. Using these methods, we investigated the effects of the sulphated octapeptide of cholecystokinin (CCK8) on gastric emptying and on stomach motility in vivo and in vitro. Gastric emptying of 99Tcm-labelled food was measured in swimming fish by using a gamma camera, counting consecutive 2.5 min periods for 18-42 h. After 20 h, 55.3+/-4.0 % of the labelled food remained in the stomach of the control fish (mean s.e.m., N=9). Vascular infusion of CCK8 (25 pmol kg-1 h-1) delayed gastric emptying so that 70.4+/-4.8 % of the labelled food remained in the stomach after 20 h (N=8). Gastric pressure changes in vivo were measured using a balloon surgically fitted into the cardiac or pyloric part of the stomach. In the cardiac part, intra arterial infusion of CCK8 at 0.1 nmol kg-1 h-1 resulted in a decrease in the frequency and amplitude of rhythmic contractions, while higher doses started/increased contractions. Atropine blocked much of the basal contractile activity, but did not influence the CCK8-induced inhibition of contractile activity. The pyloric part of the stomach was unaffected by intra-arterial infusion of CCK8 or atropine. In vitro perfusion of the stomach (with a balloon placed in the cardiac part to record motility) with CCK8 at high concentrations (10(-7 )mol l-1 and above) augmented the spontaneous contractions, while lower concentrations had inconsistent effects. In addition, CCK8 (10(-7) to 10(-6 )mol l-1) decreased the amplitude of spontaneous contractions in longitudinal strip preparations, usually in combination with an increase in the resting tension. The decrease in amplitude was not affected by the nitric oxide synthase inhibitor NG nitro-l-arginine methyl ester hydrochloride (L-NAME; 10(-4 )mol l-1). Depending on the concentration and experimental arrangement, CCK8 had either inhibitory or excitatory effects on the cardiac stomach, suggesting the possible presence of different types of CCK receptor. We conclude that the predominant effect of CCK8 in vivo may be a slowing down of gastric emptying, presumably coinciding with a release of bile into the duodenum. PMID- 9851906 TI - Control of abdominal extension in the freely moving intact crayfish cherax destructor. I. Activity Of the tonic stretch receptor AB - Electrical recordings were made from the sensory neurone of the tonic muscle receptor organ in the abdomen of the intact, freely behaving crayfish Cherax destructor. Slow extensions of the abdomen were evoked by lowering a platform from beneath the suspended crayfish, and the movements and tonic sensory neurone activity were video-recorded simultaneously. The recordings showed that the tonic sensory neurone was active when the abdomen was fully flexed prior to the extension. When the extension began, however, the sensory neurone ceased firing shortly after movement was detected, irrespective of the load applied to the abdomen. When the abdomen was physically blocked from extending fully, the sensory neurone did not fire. The tonic muscle receptor organ is considered to be the length-detecting sensor for a load-compensating servo-loop, but the results demonstrate that its activity pattern during extensions evoked by a platform-drop in C. destructor are not consistent with that role. PMID- 9851907 TI - Control of abdominal extension in the freely moving intact crayfish cherax destructor. II. Activity Of the superficial extensor motor neurones AB - The activity of the superficial extensor motor neurones was recorded during slow abdominal extension in the crayfish Cherax destructor. Postural extensions were evoked by lowering a platform from beneath the suspended crayfish. During extensions where the abdomen was physically blocked from achieving full extension, the largest superficial extensor motor neurone (SEMN6) fired at a higher rate than during unhindered extensions. Blocking a segment neighbouring that being examined also increased SEMN6 activity, demonstrating an intersegmental spread of the reflex. The increase in SEMN6 firing rate occurred in the absence of activity in the sensory neurone of the tonic muscle receptor organ, demonstrating that the tonic sensory neurone is not necessary for load compensation during these abdominal extensions in C. destructor. The findings support earlier evidence suggesting that other receptor systems can mediate load compensation in the abdomen of the crayfish. PMID- 9851908 TI - Escape trajectories of the brown shrimp crangon crangon, and a theoretical consideration of initial escape angles from predators AB - Tail-flip escape trajectories of the brown shrimp Crangon crangon have been investigated in response to a natural predator, the cod Gadus morhua, and an artificial stimulus. Shrimps escaped by rolling to their left or right during the initial tail-flip of a response, and thereafter swam on their side. As a result of the laterally directed first tail-flip, initial escape angles always lay between 75 degrees and 156 degrees with respect to the (pre-escape) longitudinal axis (anterior=0 degrees) of the shrimp. Symmetrical attacks from either head-on or tail-on produced escapes to the shrimp's left or right in equal proportions, although a contralateral bias did occur if the shrimp experienced a looming object from one side before a symmetrical attack was applied. Lateral attacks produced a significantly greater proportion of contralateral responses than ipsilateral ones. Empirical and theoretical analyses indicate that the initial escape direction is influenced by an interaction between the range of first tail flip escape angles that the shrimp is capable of performing and the risk of being intercepted by a predator during the initial stage of an escape. Thus, the unpredictability ('protean behaviour') of the response may be affected by the conditions of the interaction. Subsequent tail-flips of an escape usually directed the response away from the stimulus, but sometimes escapes were instead steered to the side of the stimulus and then behind it. The probability of each type of escape occurring changed with attack direction. The elements of protean behaviour that have been identified in both the initial and subsequent stages of the escape may prevent predators from learning a fixed pattern of response, but a trade-off occurs when escape trajectories infringe upon zones of high capture risk. PMID- 9851909 TI - The self-referencing oxygen-selective microelectrode: detection of transmembrane oxygen flux from single cells. AB - A self-referencing, polarographic, oxygen-selective microelectrode was developed for measuring oxygen fluxes from single cells. This technique is based on the translational movement of the microelectrode at a known frequency through an oxygen gradient, between known points. The differential current of the electrode was converted into a directional measurement of flux using the Fick equation. Operational characteristics of the technique were determined using artificial gradients. Calculated oxygen flux values matched theoretical values derived from static measurements. A test preparation, an isolated neuron, yielded an oxygen flux of 11.46+/-1.43 pmol cm-2 s-1 (mean +/- s.e.m.), a value in agreement with those available in the literature for single cells. Microinjection of metabolic substrates or a metabolic uncoupler increased oxygen flux, whereas microinjection of KCN decreased oxygen flux. In the filamentous alga Spirogyra greveilina, the probe could easily differentiate a 16.6% difference in oxygen flux with respect to the position of the spiral chloroplast (13.3+/-0.4 pmol cm-2 s-1 at the chloroplast and 11.4+/-0.4 pmol cm-2 s-1 between chloroplasts), despite the fact that these positions averaged only 10.6+/-1.8 microm apart (means +/- s.e.m.). A light response experiment showed real-time changes in measured oxygen flux correlated with changes in lighting. Taken together, these results show that the self-referencing oxygen microelectrode technique can be used to detect local oxygen fluxes with a high level of sensitivity and spatial resolution in real time. The oxygen fluxes detected reliably correlated with the metabolic state of the cell. PMID- 9851910 TI - The Alzheimer's plaques, tangles and memory deficits may have a common origin. Part IV: can calpain act as alpha-secretase? AB - Abnormality of protease activities and imbalance of intracellular calcium are two most salient aberrant events in Alzheimer's disease (AD). As such, calcium dependent proteases such as calpain, as a critical link between these two events, must play a key role in the pathogenesis of AD, particularly in the abnormal processing of beta-amyloid precursor protein. Because alpha-secretase in this process appears to be a calcium-dependent protease and its enzymatic characteristics are impressively similar to those of calpain, a challenging possibility arises: Calpain might act as alpha-secretase in vivo. However, as the experimental evidence both for and against this possibility is compelling, the issue currently remains as a theoretical dilemma in which a central question is whether calpain, a cytosolic enzyme, can somehow reach the cell surface. This difficult issue needs to be addressed now. As a first attempt to explore the issue, we propose a working model for the membrane orientation of calpain and suggest several experiments that will critically test this model. The quest to this dilemma will not only impact our understanding of AD, but may also expand the current knowledge about Ca2+ signal transduction pathway. Finally, we discuss several competing models and the potential role of presenilins as "regulators" of alpha-secretase. It is of interest to note that some of our previous theoretical predictions have been experimentally observed. PMID- 9851911 TI - The outer parts of the mycobacterial envelope as permeability barriers. AB - The permeability of mycobacteria to substances in their environment is controlled by the properties of their envelopes. Two special features are important: an outer lipid barrier based on a monolayer of characteristic mycolic acids and a capsule-like coat of polysaccharide and protein. The mycolate layer prevents entry of small hydrophilic molecules, which obtain access to the cell by way of pore-forming proteins resembling porins of Gram-negative bacteria. More lipophilic molecules can diffuse through the lipid layer. The capsule probably impedes access by macromolecules; in intracellular pathogenic species it forms the electron-transparent zone that separates the bacterium from the membrane of the host phagosome. The structure of the outer lipid barrier seems common to all mycobacteria, fast- and slow-growing, but the capsule is more abundant in slow growing species, a group which includes all the important mycobacterial pathogens. Mycobacteria secrete proteins into their environment, which are likely to be important in the pathogenesis of mycobacterial diseases. Knowledge of how these proteins, and the polysaccharides of the capsule, cross the outer lipid barrier is minimal at present. It is likely that proper knowledge of mycobacterial permeability will enable new approaches to treatment of mycobacterial disease. PMID- 9851912 TI - The clinical significance of sperm-zona pellucida binding. AB - The development of homologous functional bio-assay for sperm quality assessment has been a focal point of reproductive biologists; in order to provide a scientific based diagnosis in cases of fertilization failure. The availability of oocytes still remains an important limiting factor for laboratories to embark on the methodology of the assay. The use of zonae pellucidae, derived from post mortem and different in vitro fertilization oocytes, enhanced to availability of zonae. Sperm-zona binding has been illustrated to be an essential requisite during human fertilization and can be measured under hemizona assay as well intact zona pellucida conditions. The sensitivity and specificity of sperm-zona binding results indicated the assay to be positively and significantly correlated with in vitro fertilization outcome. Furthermore, a highly significant correlation were illustrated to exist between the normal sperm morphology, hyperactivation, sperm creatine kinase activity and the zona binding capacity of a given sperm sample. It was concluded that andrology testing remains an ever growing component in the work-up of the infertile couple. We enter the next millennium with many questions that remain to be answered by the hand of efficacious screening techniques and a new formidable therapy in intra cellular sperm injection. PMID- 9851913 TI - Neuroendocrine and immune mediators in prostate cancer progression. AB - Cytokines constitute a diverse group of intercellular signaling proteins that regulate local and systemic, immune and inflammatory responses as well as wound healing and hematopoiesis. The proliferation and maturation of cells of the immune system, both normal and malignant, is regulated by cytokines such as the interleukins. Such cytokines may also influence the proliferation and differentiation of other cell types. Prostate epithelial cells differentiate along two pathways, exocrine or neuroendocrine. Elevation in the exocrine marker prostate-specific antigen and/or the neuroendocrine marker chromogranin A in serum has been associated with prostate cancer progression. Interleukin-1 (IL-1) mRNA is expressed by two androgen-insensitive (AI) but not by three androgen sensitive prostate cancer cell lines. IL-1 inhibits while IL-2 stimulates the growth of the androgen-sensitive LNCaP cell line. Neither affects growth of AI PC 3 or DU-145 cell lines. IL-1 promotes the neuroendocrine phenotype and IL-2 promotes the exocrine phenotype in prostate cancer. The influence of the immune mediators IL-1 and IL-2 on the growth and differentiation of prostate cancer cells and its implication in tumor progression is described herein. Relationship of IL-1 with bone metastasis and the involvement of ss-2 microglobulin in the development and progression of prostate cancer are also discussed. PMID- 9851914 TI - Role of PP2A in intracellular signal transduction pathways. AB - Intracellular signal transduction pathways play a crucial role in a variety of cellular processes, such as differentiation, proliferation, or apoptosis, and the reversible phosphorylation of their components is a major regulatory mechanism to control their activities. While much has been learned about the contribution of kinases, the involvement of phosphatases in these events is less clear and has only recently received more investigative attention. The availability of various natural product inhibitors of phosphatases has helped enormously to gain insight into the role that these enzymes exert in various signal transduction processes. This review will focus on serine/threonine protein phosphatase type 2A (PP2A) and will present findings pertaining to its involvement in cellular signal transduction pathways. Since the majority of these studies were done with the use of phosphatase inhibitory compounds, some pros and cons of their application will be presented. PMID- 9851915 TI - Treatment of severe male infertility by micromanipulation-assisted fertilization: news and views. AB - Recent progress in cell micromanipulation has made it possible to achieve fertilization even with highly deficient male gametes, by injecting them directly to oocyte cytoplasm. This technique, known as intracytoplasmic sperm injection (ICSI), has proved efficient in most types of sperm pathology. However, ICSI fails when injected spermatozoa are not capable of supporting the process of oocyte activation or when they carry genetic abnormalities incompatible with normal embryonic development. Abnormalities of oocyte activation and the transmission of genetic anomalies to the progeny are also two major concerns relating to the clinical use of ICSI. The use of micromanipulation-assisted fertilization in cases of male germ cell maturation arrest, namely fertilization by round spermatid injection (ROSI), has brought preliminary clinical results, and the improvement of ROSI clinical efficacy remains a major challenge for future research. PMID- 9851916 TI - Genome sequence of the nematode C. elegans: a platform for investigating biology. AB - The 97-megabase genomic sequence of the nematode Caenorhabditis elegans reveals over 19,000 genes. More than 40 percent of the predicted protein products find significant matches in other organisms. There is a variety of repeated sequences, both local and dispersed. The distinctive distribution of some repeats and highly conserved genes provides evidence for a regional organization of the chromosomes. PMID- 9851917 TI - Zinc fingers in Caenorhabditis elegans: finding families and probing pathways. AB - More than 3 percent of the protein sequences inferred from the Caenorhabditis elegans genome contain sequence motifs characteristic of zinc-binding structural domains, and of these more than half are believed to be sequence-specific DNA binding proteins. The distribution of these zinc-binding domains among the genomes of various organisms offers insights into the role of zinc-binding proteins in evolution. In addition, the complete genome sequence of C. elegans provides an opportunity to analyze, and perhaps predict, pathways of transcriptional regulation. PMID- 9851919 TI - Neurobiology of the Caenorhabditis elegans genome. AB - Neurotransmitter receptors, neurotransmitter synthesis and release pathways, and heterotrimeric GTP-binding protein (G protein)-coupled second messenger pathways are highly conserved between Caenorhabditis elegans and mammals, but gap junctions and chemosensory receptors have independent origins in vertebrates and nematodes. Most ion channels are similar to vertebrate channels but there are no predicted voltage-activated sodium channels. The C. elegans genome encodes at least 80 potassium channels, 90 neurotransmitter-gated ion channels, 50 peptide receptors, and up to 1000 orphan receptors that may be chemoreceptors. For many gene families, C. elegans has both conventional members and divergent outliers with weak homology to known genes; these outliers may provide insights into previously unknown functions of conserved protein families. PMID- 9851918 TI - Comparison of the complete protein sets of worm and yeast: orthology and divergence. AB - Comparative analysis of predicted protein sequences encoded by the genomes of Caenorhabditis elegans and Saccharomyces cerevisiae suggests that most of the core biological functions are carried out by orthologous proteins (proteins of different species that can be traced back to a common ancestor) that occur in comparable numbers. The specialized processes of signal transduction and regulatory control that are unique to the multicellular worm appear to use novel proteins, many of which re-use conserved domains. Major expansion of the number of some of these domains seen in the worm may have contributed to the advent of multicellularity. The proteins conserved in yeast and worm are likely to have orthologs throughout eukaryotes; in contrast, the proteins unique to the worm may well define metazoans. PMID- 9851920 TI - The taxonomy of developmental control in Caenorhabditis elegans. AB - The Caenorhabditis elegans genome sequence was surveyed for transcription factor and signaling gene families that have been shown to regulate development in a variety of species. About 10 to 25 percent of the genes in most of the gene families already have been genetically analyzed in C. elegans, about half of the genes detect probable orthologs in other species, and about 10 to 25 percent of the genes are, at present, unique to C. elegans. Caenorhabditis elegans is also missing genes that are found in vertebrates and other invertebrates. Thus the genome sequence reveals universals in developmental control that are the legacy of metazoan complexity before the Cambrian explosion, as well as genes that have been more recently invented or lost in particular phylogenetic lineages. PMID- 9851921 TI - Caenorhabditis elegans is a nematode. AB - Caenorhabditis elegans is a rhabditid nematode. What relevance does this have for the interpretation of the complete genome sequence, and how will it affect the exploitation of the sequence for scientific and social ends? Nematodes are only distantly related to humans and other animal groups; will this limit the universality of the C. elegans story? Many nematodes are parasites; can knowledge of the C. elegans sequence aid in the prevention and treatment of disease? PMID- 9851922 TI - Observations of the north polar region of Mars from the Mars orbiter laser altimeter. AB - Elevations from the Mars Orbiter Laser Altimeter (MOLA) have been used to construct a precise topographic map of the martian north polar region. The northern ice cap has a maximum elevation of 3 kilometers above its surroundings but lies within a 5-kilometer-deep hemispheric depression that is contiguous with the area into which most outflow channels emptied. Polar cap topography displays evidence of modification by ablation, flow, and wind and is consistent with a primarily H2O composition. Correlation of topography with images suggests that the cap was more spatially extensive in the past. The cap volume of 1.2 x 10(6) to 1.7 x 10(6) cubic kilometers is about half that of the Greenland ice cap. Clouds observed over the polar cap are likely composed of CO2 that condensed out of the atmosphere during northern hemisphere winter. Many clouds exhibit dynamical structure likely caused by the interaction of propagating wave fronts with surface topography. PMID- 9851923 TI - A 3.3-Ma impact in argentina and possible consequences AB - Enigmatic glassy materials (escorias) and red bricklike materials (tierras cocidas) occur at a restricted stratigraphic level (the top of the Chapadmalal Formation). Materials from one locality near Mar del Plata are attributed to a mid-Pliocene impact event with a radiometric and magnetostratigraphic age of 3.3 million years ago (Ma). An extinction of endemic fauna (including the glyptodonts and flightless cariamid birds) correlates with the unit containing the impact glasses. Moreover, the age of the glasses is coincident within dating uncertainties with a pulselike change in the oxygen isotope marine record in the Atlantic and Pacific Oceans just before the late Pliocene deterioration of the climate. PMID- 9851924 TI - The dusty atmosphere of the brown dwarf Gliese 229B. AB - The brown dwarf Gliese 229B has an observable atmosphere too warm to contain ice clouds like those on Jupiter and too cool to contain silicate clouds like those on low-mass stars. These unique conditions permit visibility to higher pressures than possible in cool stars or planets. Gliese 229B's 0.85- to 1.0-micrometer spectrum indicates particulates deep in the atmosphere (10 to 50 bars) having optical properties of neither ice nor silicates. Their reddish color suggests an organic composition characteristic of aerosols in planetary stratospheres. The particles' mass fraction (10(-7)) agrees with a photochemical origin caused by incident radiation from the primary star and suggests the occurrence of processes native to planetary stratospheres. PMID- 9851925 TI - Photoemission evidence for a remnant fermi surface and a d-wave-like dispersion in insulating Ca2CuO2Cl2 AB - An angle-resolved photoemission study is reported on Ca2CuO2Cl2, a parent compound of high-Tc superconductors. Analysis of the electron occupation probability, n(k), from the spectra shows a steep drop in spectral intensity across a contour that is close to the Fermi surface predicted by the band calculation. This analysis reveals a Fermi surface remnant, even though Ca2CuO2Cl2 is a Mott insulator. The lowest energy peak exhibits a dispersion with approximately the &cjs3539;coskxa - coskya&cjs3539; form along this remnant Fermi surface. Together with the data from Dy-doped Bi2Sr2CaCu2O8+delta, these results suggest that this d-wave-like dispersion of the insulator is the underlying reason for the pseudo gap in the underdoped regime. PMID- 9851926 TI - Promotion of trophoblast stem cell proliferation by FGF4. AB - The trophoblast cell lineage is essential for the survival of the mammalian embryo in utero. This lineage is specified before implantation into the uterus and is restricted to form the fetal portion of the placenta. A culture of mouse blastocysts or early postimplantation trophoblasts in the presence of fibroblast growth factor 4 (FGF4) permitted the isolation of permanent trophoblast stem cell lines. These cell lines differentiated to other trophoblast subtypes in vitro in the absence of FGF4 and exclusively contributed to the trophoblast lineage in vivo in chimeras. PMID- 9851927 TI - Role of alpha-dystroglycan as a Schwann cell receptor for Mycobacterium leprae. AB - alpha-Dystroglycan (alpha-DG) is a component of the dystroglycan complex, which is involved in early development and morphogenesis and in the pathogenesis of muscular dystrophies. Here, alpha-DG was shown to serve as a Schwann cell receptor for Mycobacterium leprae, the causative organism of leprosy. Mycobacterium leprae specifically bound to alpha-DG only in the presence of the G domain of the alpha2 chain of laminin-2. Native alpha-DG competitively inhibited the laminin-2-mediated M. leprae binding to primary Schwann cells. Thus, M. leprae may use linkage between the extracellular matrix and cytoskeleton through laminin-2 and alpha-DG for its interaction with Schwann cells. PMID- 9851928 TI - Identification of alpha-dystroglycan as a receptor for lymphocytic choriomeningitis virus and Lassa fever virus. AB - A peripheral membrane protein that is interactive with lymphocytic choriomeningitis virus (LCMV) was purified from cells permissive to infection. Tryptic peptides from this protein were determined to be alpha-dystroglycan (alpha-DG). Several strains of LCMV and other arenaviruses, including Lassa fever virus (LFV), Oliveros, and Mobala, bound to purified alpha-DG protein. Soluble alpha-DG blocked both LCMV and LFV infection. Cells bearing a null mutation of the gene encoding DG were resistant to LCMV infection, and reconstitution of DG expression in null mutant cells restored susceptibility to LCMV infection. Thus, alpha-DG is a cellular receptor for both LCMV and LFV. PMID- 9851929 TI - Proofreading and aminoacylation of tRNAs before export from the nucleus. AB - After synthesis and processing in the nucleus, mature transfer RNAs (tRNAs) are exported to the cytoplasm in a Ran.guanosine triphosphate-dependent manner. Export of defective or immature tRNAs is avoided by monitoring both structure and function of tRNAs in the nucleus, and only tRNAs with mature 5' and 3' ends are exported. All tRNAs examined can be aminoacylated in nuclei of Xenopus oocytes, thereby providing a possible mechanism for functional proofreading of newly made tRNAs. Inhibition of aminoacylation of a specific tRNA retards its appearance in the cytoplasm, indicating that nuclear aminoacylation promotes efficient export. PMID- 9851930 TI - Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. AB - Mutations of the gene Lps selectively impede lipopolysaccharide (LPS) signal transduction in C3H/HeJ and C57BL/10ScCr mice, rendering them resistant to endotoxin yet highly susceptible to Gram-negative infection. The codominant Lpsd allele of C3H/HeJ mice was shown to correspond to a missense mutation in the third exon of the Toll-like receptor-4 gene (Tlr4), predicted to replace proline with histidine at position 712 of the polypeptide chain. C57BL/10ScCr mice are homozygous for a null mutation of Tlr4. Thus, the mammalian Tlr4 protein has been adapted primarily to subserve the recognition of LPS and presumably transduces the LPS signal across the plasma membrane. Destructive mutations of Tlr4 predispose to the development of Gram-negative sepsis, leaving most aspects of immune function intact. PMID- 9851931 TI - Exploiting the basis of proline recognition by SH3 and WW domains: design of N substituted inhibitors. AB - Src homology 3 (SH3) and WW protein interaction domains bind specific proline rich sequences. However, instead of recognizing critical prolines on the basis of side chain shape or rigidity, these domains broadly accepted amide N-substituted residues. Proline is apparently specifically selected in vivo, despite low complementarity, because it is the only endogenous N-substituted amino acid. This discriminatory mechanism explains how these domains achieve specific but low affinity recognition, a property that is necessary for transient signaling interactions. The mechanism can be exploited: screening a series of ligands in which key prolines were replaced by nonnatural N-substituted residues yielded a ligand that selectively bound the Grb2 SH3 domain with 100 times greater affinity. PMID- 9851932 TI - Defective T cell differentiation in the absence of Jnk1. AB - The c-Jun NH2-terminal kinase (JNK) signaling pathway has been implicated in the immune response that is mediated by the activation and differentiation of CD4 helper T (TH) cells into TH1 and TH2 effector cells. JNK activity observed in wild-type activated TH cells was severely reduced in TH cells from Jnk1-/- mice. The Jnk1-/- T cells hyperproliferated, exhibited decreased activation-induced cell death, and preferentially differentiated to TH2 cells. The enhanced production of TH2 cytokines by Jnk1-/- cells was associated with increased nuclear accumulation of the transcription factor NFATc. Thus, the JNK1 signaling pathway plays a key role in T cell receptor-initiated TH cell proliferation, apoptosis, and differentiation. PMID- 9851933 TI - Eight calves cloned from somatic cells of a single adult. AB - Eight calves were derived from differentiated cells of a single adult cow, five from cumulus cells and three from oviductal cells out of 10 embryos transferred to surrogate cows (80 percent success). All calves were visibly normal, but four died at or soon after birth from environmental causes, and postmortem analysis revealed no abnormality. These results show that bovine cumulus and oviductal epithelial cells of the adult have the genetic content to direct the development of newborn calves. PMID- 9851934 TI - Elevating the vitamin E content of plants through metabolic engineering. AB - alpha-Tocopherol (vitamin E) is a lipid-soluble antioxidant synthesized only by photosynthetic organisms. alpha-Tocopherol is an essential component of mammalian diets, and intakes in excess of the U.S. recommended daily allowance are correlated with decreased incidence of a number of degenerative human diseases. Plant oils, the main dietary source of tocopherols, typically contain alpha tocopherol as a minor component and high levels of its biosynthetic precursor, gamma-tocopherol. A genomics-based approach was used to clone the final enzyme in alpha-tocopherol synthesis, gamma-tocopherol methyltransferase. Overexpression of gamma-tocopherol methyltransferase in Arabidopsis seeds shifted oil compositions in favor of alpha-tocopherol. Similar increases in agricultural oil crops would increase vitamin E levels in the average U.S. diet. PMID- 9851935 TI - Pathophysiological role of angiotensin II type 2 receptor in cardiovascular and renal diseases. AB - Since the discovery of nonpeptidic ligands, the receptors for angiotensin (Ang) II have been classified into 2 subtypes (Ang II type 1 receptor [AT1-R] and Ang II type 2 receptor [AT2-R]). AT1-R mediates most of the cardiovascular actions of Ang II. AT2-R is expressed at very high levels in the developing fetus. Its expression is very low in the cardiovascular system of the adult. The expression of AT2-R can be modulated by pathological states associated with tissue remodeling or inflammation. In failing hearts or neointima formation after vascular injury, AT2-R is reexpressed in cells proliferating in interstitial regions or neointima and exerts an inhibitory effect on Ang II-induced mitogen signals or synthesis of extracellular matrix proteins, resulting in attenuation of the tissue remodeling. An extreme form of cell growth inhibition ends in programmed cell death, and this process, which is initiated by the withdrawal of growth factors, is also enhanced by AT2-R. Cardiac myocyte- or vascular smooth muscle-specific mice that overexpress AT2-R display an inhibition of Ang II induced chronotropic or pressor actions, suggesting the role of AT2-R on the activity of cardiac pacemaker cells and the maintenance of vascular resistance. AT2-R also activates the kinin/nitric oxide/cGMP system in the cardiovascular and renal systems, resulting in AT2-R-mediated cardioprotection, vasodilation, and pressure natriuresis. These effects, transmitted by AT2-R, are mainly exerted by stimulation of protein tyrosine or serine/threonine phosphatases in a Gi protein dependent manner. The expression level of AT2-R is much higher in human hearts than in rodent hearts, and the AT2-R-mediated actions are likely enhanced, especially by clinical application of AT1-R antagonists. Thus, in this review, the regulation of AT2-R expression, its cellular localization, its pathological role in cardiovascular and kidney diseases, and pharmacotherapeutic effects of AT2-R stimulation are discussed. PMID- 9851936 TI - Bay K 8644 increases resting Ca2+ spark frequency in ferret ventricular myocytes independent of Ca influx: contrast with caffeine and ryanodine effects. AB - Bay K 8644, an L-type Ca2+ channel agonist, was shown previously to increase resting sarcoplasmic reticulum (SR) Ca2+ loss and convert post-rest potentiation to decay in dog and ferret ventricular muscle. Here, the effects of Bay K 8644 on local SR Ca2+ release events (Ca2+ sparks) were measured in isolated ferret ventricular myocytes, using laser scanning confocal microscopy and the fluorescent Ca2+ indicator fluo-3. The spark frequency under control conditions was fairly constant during 20 s of rest after interruption of electrical stimulation. Bay K 8644 (100 nmol/L) increased the spark frequency by 466+/-90% of control at constant SR Ca2+ load but did not change the spatial and temporal characteristics of individual sparks. The increase in spark frequency was maintained throughout the period of rest. The increase in Ca2+ spark frequency induced by Bay K 8644 was not affected by superfusion with Ca2+-free solution (with 10 mmol/L EGTA) but was suppressed by the addition of 10 micromol/L nifedipine (which by itself did not alter resting Ca2+ spark frequency). This suggests that the effect of Bay K 8644 on Ca2+ sparks is mediated by the sarcolemmal dihydropyridine receptor but is also independent of Ca2+ influx. Low concentrations of caffeine (0.5 mmol/L) increased both the average frequency and duration of sparks. Ryanodine (50 nmol/L) increased the spark frequency and also induced long-lasting Ca2+ signals. This may indicate long-lasting openings of SR Ca2+ release channels and a lack of local SR Ca2+ depletion. In lipid bilayers, Bay K 8644 had no effect on either single-channel current amplitude or open probability of the cardiac ryanodine receptor. It is concluded that Bay K 8644 activates SR Ca2+ release at rest, independent of Ca2+ influx and perhaps through a functional linkage between the sarcolemmal dihydropyridine receptor and the SR ryanodine receptor. In contrast, caffeine and ryanodine modulate Ca2+ sparks by a direct action on the SR Ca2+ release channels. PMID- 9851937 TI - Targeted overexpression of the sarcoplasmic reticulum Ca2+-ATPase increases cardiac contractility in transgenic mouse hearts. AB - Cardiac hypertrophy and heart failure are known to be associated with a reduction in Ca2+-ATPase pump levels of the sarcoplasmic reticulum (SR). To determine whether, and to what extent, alterations in Ca2+ pump numbers can affect contraction and relaxation parameters of the heart, we have overexpressed the cardiac SR Ca2+-ATPase specifically in the mouse heart using the alpha-myosin heavy chain promoter. Analysis of 2 independent transgenic lines demonstrated that sarco(endo)plasmic reticulum Ca2+-ATPase isoform (SERCA2a) mRNA levels were increased 3.88+/-0. 4-fold and 7.90+/-0.2-fold over those of the control mice. SERCA2a protein levels were increased by 1.31+/-0.05-fold and 1.54+/-0. 05-fold in these lines despite high levels of mRNA, suggesting that complex regulatory mechanisms may determine the SERCA2a pump levels. The maximum velocity of Ca2+ uptake (Vmax) was increased by 37%, demonstrating that increased pump levels result in increased SR Ca2+ uptake function. However, the apparent affinity of the SR Ca2+-ATPase for Ca2+ remains unchanged in transgenic hearts. To evaluate the effects of overexpression of the SR Ca2+ pump on cardiac contractility, we used the isolated perfused work-performing heart model. The transgenic hearts showed significantly higher myocardial contractile function, as indicated by increased maximal rates of pressure development for contraction (+dP/dt) and relaxation (-dP/dt), together with shortening of the normalized time to peak pressure and time to half relaxation. Measurements of intracellular free calcium concentration and contractile force in trabeculae revealed a doubling of Ca2+ transient amplitude, with a concomitant boost in contractility. The present study demonstrates that increases in SERCA2a pump levels can directly enhance contractile function of the heart by increasing SR Ca2+ transport. PMID- 9851938 TI - Overexpression of the cardiac Na+/Ca2+ exchanger increases susceptibility to ischemia/reperfusion injury in male, but not female, transgenic mice. AB - Influx of Ca2+ into myocytes via Na+/Ca2+ exchange may be stimulated by the high levels of intracellular Na+ and the changes in membrane potential known to occur during ischemia/reperfusion. This increased influx could, in turn, lead to Ca2+ overload and injury. Overexpression of the cardiac Na+/Ca2+ exchanger therefore may increase susceptibility to ischemia/reperfusion injury. To test this hypothesis, the hearts of male and female transgenic mice, overexpressing the Na+/Ca2+ exchange protein, and hearts of their wild-type littermates, were perfused with Krebs-Henseleit buffer and subjected to 20 minutes of ischemia and 40 minutes of reperfusion. Preischemic left ventricular developed pressures and +dP/dtmax, as well as -dP/dtmin, were higher in the male transgenic hearts compared with wild-type, implying a role for Na+/Ca2+ exchange in the contraction, as well as the relaxation, phases of the cardiac beat. Postischemic function was lower in male transgenic than in male wild-type hearts (7+/-2% versus 32+/-6% of preischemic function), but there was no difference between female transgenic and female wild-type hearts, both at approximately 30% of preischemic function. To assess whether this male/female difference was due to female-specific hormones such as estrogen, the hearts of bilaterally ovariectomized and sham-operated transgenic females were subjected to the same protocol. The functional recoveries of ovariectomized female transgenic hearts were lower (17+/-3% of preischemic function) than those of wild-type and sham operated transgenic females. The lower postischemic functional recovery in the male transgenic and female ovariectomized transgenic hearts correlated with lower recoveries of the energy metabolites, ATP and phosphocreatine, as measured by 31P nuclear magnetic resonance spectroscopy. Alternans were observed during reperfusion in male transgenic and female ovariectomized transgenic hearts only, consistent with intracellular Ca2+ overload. Western analyses showed that alterations in the expression of the Na+/Ca2+ exchange or L-type Ca2+ channel proteins were not responsible for the protection observed in the female transgenic hearts. In conclusion, in males, overexpression of the Na+/Ca2+ exchanger reduced postischemic recovery of both contractile function and energy metabolites, indicating that the Na+/Ca2+ exchanger may play a role in ischemia/reperfusion injury. From the studies of females, however, it appears that this exacerbation of ischemia/reperfusion injury by overexpression of the Na+/Ca2+ exchanger can be overcome partially by female-specific hormones such as estrogen. PMID- 9851939 TI - Quinidine interactions with human atrial potassium channels: developmental aspects. AB - Clinical studies have suggested that quinidine is less effective when used for the treatment of atrial arrhythmias in pediatric patients compared with its clinical effectiveness in the adult patient population. Age-related changes in the cardiac actions of quinidine on action potential duration and interaction with potassium channels in several mammalian species also have been reported. We investigated the effects of postnatal development on quinidine's interaction with major repolarizing currents (Ito, IKur, Ins, and IK1) in human atrial myocytes, using the whole-cell configuration of the voltage-clamp technique. Our results indicate that there are age-related changes in both the IC50 for quinidine blockade of Ito, as well as the mechanism of quinidine unblocking. In contrast, quinidine was found to inhibit both adult and pediatric IK1 and IKur in an age independent manner, whereas the nonselective cation current (Ins), which contributes to the sustained outward current (Isus), was insensitive to quinidine. The results from this study help to clarify the electrophysiological mechanism by which quinidine elicits its antiarrhythmic effect in the pediatric and adult human population. PMID- 9851940 TI - Reduced reperfusion-induced Ins(1,4,5)P3 generation and arrhythmias in hearts expressing constitutively active alpha1B-adrenergic receptors. AB - Reperfusion of globally ischemic rat hearts causes the generation of inositol(1,4,5)trisphosphate [Ins(1,4,5)P3] and the initiation of arrhythmias. These responses are mediated by alpha1-adrenergic receptors (ARs), but the subtype of receptor involved has not been identified. Under normoxic conditions, hearts from transgenic animals expressing constitutively active alpha1B-ARs in heart (alpha1B-constitutively active mutant [CAM]) showed higher [3H] inositol phosphate responses to norepinephrine (2.3-fold) than hearts from nontransgenic animals (alpha1B-WT) (1.6-fold). alpha1B-WT hearts responded to 2 minutes of reperfusion after 20 minutes of global ischemia by generation of Ins(1,4,5)P3 (5301+/-1310 to 11 413+/-1597 CPM/g tissue; mean+/-SEM; n=6; P<0.01 in [3H] labeling studies and 3.8+/-0.2 to 6.3+/-0.6 nmol/g by mass analysis, n=6; P<0.05). In contrast to findings in normoxia, hearts from alpha1B-CAM animals showed no Ins(1,4,5)P3 response in early reperfusion. In parallel studies, alpha1B-WT hearts developed ventricular tachycardia and ventricular premature beats (VPB) during 5 minutes of reperfusion after 20 minutes of ischemia. The incidence of these arrhythmias was reduced in the alpha1B-CAM hearts (95% to 62% for VPB and 47% to 12% for ventricular tachycardia; both P<0.05). The resistance of the alpha1B-CAM hearts was not due to alpha1B-AR-mediated preconditioning, as the Ins(1,4,5)P3 response to thrombin receptor activation during reperfusion was not different between the 2 groups. To investigate the possibility of reduced alpha1A-receptor activity in the alpha1B-CAM hearts, expression of the mRNA for alpha1A- and alpha1B-receptors was measured. alpha1B-WT hearts contained mRNA for both receptor subtypes, but the levels of alpha1B-receptor mRNA were 5-fold higher than alpha1A-receptor mRNA. alpha1B-CAM hearts contained very high levels of alpha1B-receptor mRNA (26-fold increase), but the expression of mRNA for the alpha1A-receptors (0.141+/-0.035 amol/ microg RNA; mean+/-SEM; n=6) was reduced by 50% relative to alpha1B-WT controls (0.276+/-0.046 amol/ microg RNA; n=6; P<0.01). The reduction in arrhythmogenic and Ins(1,4,5)P3 responses in alpha1B CAM hearts provides evidence that these response are not mediated by alpha1B receptors. PMID- 9851941 TI - Nicotine enhances presynaptic and postsynaptic glutamatergic neurotransmission to activate cardiac parasympathetic neurons. AB - Although peripheral cholinergic neurotransmission has long been known to play a pivotal role in the control of heart rate and blood pressure, recent evidence has suggested that central cholinergic mechanisms may be involved in the genesis of hypertension, anxiety, cardiorespiratory control, and, in particular, the respiratory modulation of heart rate. Yet, the sites, mechanisms, and receptor subtypes involved in the action of nicotine within the central nervous system are controversial. The present study demonstrates that nicotine has at least 3 sites of action to increase the activity of vagal cardiac neurons. Nicotine, but not muscarinic agonists, activates postsynaptic receptors and a depolarizing inward current in vagal cardiac neurons studied with the perforated patch-clamp technique in a visualized brain stem slice. In addition, nicotine acts at different presynaptic and postsynaptic sites to facilitate glutamatergic neurotransmission. Presynaptic nicotinic receptors increase the frequency of transmitter release and are sensitive to block by alpha-bungarotoxin. Nicotine also elicits a previously undescribed augmentation of postsynaptic non-NMDA currents. The presynaptic and postsynaptic receptors may prove to be future targets in the search for agonists to increase vagal cardiac activity and reduce the fatality associated with cardiac hyperexcitability and for antagonists to reduce cardiac vagal activity in pathological conditions associated with abnormally low heart rates and cardiac function such as sudden infant death syndrome. PMID- 9851942 TI - Individual gap junction plaques contain multiple connexins in arterial endothelium. AB - Gap-junctional intercellular communication in endothelial cells is implicated in the coordination of growth, migration, and vasomotor responses. Up to 3 connexin types, connexin40 (Cx40), Cx37, and Cx43 may be expressed in vascular endothelium according to vascular site, species, and physiological conditions. To establish how these connexins are organized at the level of the individual endothelial gap junction, we used affinity-purified connexin-specific antibodies raised in 3 different species to permit double and triple immunolabeling in combination with confocal and electron microscopy. Using HeLa cells transfected with Cx37 and Cx40 for characterization, the anti-Cx37 antibody (raised in rabbit) and the anti-Cx40 antibody (raised in guinea pig) were shown to recognize single bands of 37 and 40 kDa, respectively, on Western blots and to give prominent punctate labeling at the cell borders, specifically in the corresponding transfectant. By applying these antibodies together with mouse monoclonal anti-Cx43 for double and triple immunofluorescence labeling at confocal microscopy, rat aortic and pulmonary arterial endothelia were found to express all 3 connexin types, whereas coronary artery endothelium expressed Cx40 and Cx37 but lacked Cx43. High-resolution en face confocal viewing of the aortic endothelium after double labeling demonstrated frequent colocalization of connexins, with distinct variation in the expression pattern within a given cell, where it made contact with different neighbors. Triple immunogold labeling at the electron-microscopic level revealed that aortic endothelial gap junctions commonly contain all 3 connexin types. This represents the first definitive demonstration of any cell type in vivo expressing 3 different connexins organized within the same gap-junctional plaque. PMID- 9851943 TI - Expression of tissue factor pathway inhibitor in vascular smooth muscle cells and its regulation by growth factors. AB - Tissue factor pathway inhibitor (TFPI) in vivo is thought to be synthesized mainly by endothelial cells. To date, no significant regulator of TFPI synthesis has been described. Vascular smooth muscle cells (VSMC) express tissue factor in vitro and in vivo, which may contribute to vascular thrombosis. We hypothesized that VSMC might also express TFPI. To determine this, we examined growth-arrested coronary VSMC in culture and found that VSMC secreted an amount of TFPI similar to that seen in endothelial cells. Immunohistochemistry of normal human coronary arteries showed TFPI staining throughout the media and intima of the vessel with localization to VSMC and endothelial cells. To determine regulation of TFPI expression in VSMC, we examined the effects of serum stimulation on TFPI secretion and found that FBS induced a 5-fold increase in TFPI antigen and activity levels in conditioned medium at 48 hours (P<0.001) when compared with serum-free conditions. A similar stimulatory effect was seen with 10% pooled human serum. Moreover, epidermal growth factor and platelet-derived growth factor B increased TFPI secretion by 4- to 5-fold and 2- to 3-fold, respectively (P<0.05), and these growth factors accounted for approximately 50% of the TFPI secretion effects of human serum. The serum effect was associated with a 3-fold increase in TFPI mRNA 24 hours after release from growth arrest and a 50% decrease in TFPI secretion after treatment with actinomycin D. Taken together, this study suggests that there is significant TFPI expression in VSMC in culture and in VSMC within the intima and media of the normal coronary artery wall. We present the first evidence for TFPI regulation by serum in VSMC and more specifically by its constituent growth factors, epidermal growth factor and platelet-derived growth factor-B. PMID- 9851946 TI - Corrections PMID- 9851944 TI - Neuronal nitric oxide synthase is expressed in rat vascular smooth muscle cells: activation by angiotensin II in hypertension. AB - The nitric oxide synthase (NOS) inhibitor nitro-L-arginine augmented the contractions to angiotensin (Ang) II in carotid artery rings without endothelium from spontaneously hypertensive rats (SHR) but not normotensive Wistar-Kyoto rats, suggesting the possibility of nonendothelial NOS activity in SHR arteries. In SHR artery without endothelium, the potentiation of Ang II contraction by nitro-L-arginine was prevented by L-arginine, but not by D-arginine, and was observed also in the presence of oxyhemoglobin, monomethyl-L-arginine, and 7 nitroindazole, but not in the presence of aminoguanidine. In further support of NOS activation by Ang II in nonendothelial cells, Ang II but not acetylcholine stimulated cGMP levels by 2-fold in SHR arteries without endothelium; nitro-L arginine decreased both basal and Ang II-stimulated cGMP levels. When NOS activity in SHR arteries was measured, no calcium-independent L-citrulline formation was detectable, while up to 47% of the total calcium-dependent NOS activity was present in nonendothelial cells. Expression of neuronal NOS was revealed in the media of SHR arteries by immunohistochemistry, Western blot, and reverse transcriptase-polymerase chain reaction. Expression of this NOS isoform was greater in SHR than in Wistar-Kyoto rat preparations. Finally, endothelial NOS was observed in the endothelium, but no detectable levels of inducible NOS were found in these tissues. These results demonstrate the expression of neuronal NOS in rat vascular smooth muscle cells and its activation on stimulation by Ang II in spontaneously hypertensive, but not normotensive, animals. PMID- 9851945 TI - Kidney is the only source of human plasma renin in 45-kb human renin transgenic mice. AB - Prorenin is expressed in certain extrarenal tissues, but normally only the kidneys process prorenin to renin and secrete renin into the circulation. Although transgenic animal lines containing the human renin (hREN) structural gene with either 0.9-kb or 3-kb 5'-flanking DNA express the transgene appropriately in renal juxtaglomerular cells and secrete hREN into the circulation, the source of the circulating renin is not known. In the present study, we observed that 13-kb hREN transgenic mice that contain the structural gene and 0.9-kb 5'-flanking DNA express hREN mRNA in many unusual tissues. We also observed that circulating hREN levels in 13-kb hREN mice increased after bilateral nephrectomy. These results suggested that the hREN gene is expressed at inappropriate locations where prorenin might be processed to renin. To determine if more distal sequences flanking the hREN gene might contribute to cell and tissue specificity, we used a 45-kb hREN genomic fragment that contained the structural gene and about 25-kb 5'- and 8-kb 3'-flanking DNA sequences to generate 3 separate transgenic lines that contained the intact transgene sequences. Ribonuclease protection assays revealed a much narrower tissue distribution of hREN expression than in the 13-kb hREN transgenic mice. In each 45-kb hREN line, hREN mRNA was present only in the kidney, adrenal, lung, eye, ovary, and brain. Moreover, 24 hours after nephrectomy, human plasma renin fell to very low levels, indistinguishable from those of nontransgenic littermates, indicating that their circulating hREN is of renal origin. These studies suggest that sequences flanking the structural gene, missing from previous hREN transgenic lines, suppress renin gene expression at inappropriate extrarenal sites where cellular proteases, to which prorenin is not normally exposed, could convert prorenin to renin, resulting in abnormal secretion of renin into the plasma. PMID- 9851947 TI - Ten years of benefit from a one-hour intervention. PMID- 9851948 TI - Endothelium as a therapeutic target in heart failure. PMID- 9851949 TI - Nonischemic chest pain induced by coronary interventions: a prospective study comparing coronary angioplasty and stent implantation. AB - BACKGROUND: Chest pain frequently occurs without any signs of ischemia within the first 24 hours after coronary interventions. To test the hypothesis that this pain may be due to local vessel injury ("stretch pain"), we performed a prospective study enrolling patients after PTCA, stent implantation, or diagnostic coronary angiography alone. METHODS AND RESULTS: A total of 145 patients after coronary angiography were evaluated by a validated questionnaire for quantifying postinterventional chest pain within 24 hours. To detect myocardial ischemia, all patients were evaluated with a 12-lead ECG and cardiac isoenzymes immediately after the procedure and the morning after. After stent implantation, 21 of the 51 patients (41.2%) developed chest pain, compared with 4 of the 33 patients (12.1%) undergoing PTCA and 6 of the 61 patients (9.8%) with a diagnostic angiography (P<0.001). Of these 31 patients who developed chest pain, only 3 (9.7%) felt that the pain was similar to previously experienced angina pectoris. The minimal lumen diameter after intervention was significantly larger in the stent group than in the PTCA group (3.14+/-0.75 versus 1.95+/-0.67 mm; P<0. 001). No patient had changes in the ECG compared with before intervention, but 3 patients after stent implantation had a rise in cardiac isoenzymes. No other major adverse cardiac events occurred until discharge. CONCLUSIONS: Nonischemic chest pain develops in almost half of all patients undergoing stent implantation and seems to be related to vessel overexpansion caused by the stent in the diseased vessel segment. PMID- 9851951 TI - Measurement of atherosclerotic carotid plaque size in vivo using high resolution magnetic resonance imaging. AB - BACKGROUND: Current imaging modalities, such as contrast angiography, accurately determine the degree of luminal narrowing but provide no direct information on plaque size. Magnetic resonance imaging (MRI), however, has potential for noninvasively determining arterial wall area (WA). This study was conducted to determine the accuracy of in vivo MRI for measuring the cross-sectional maximum wall area (MaxWA) of atherosclerotic carotid arteries in a group of patients undergoing carotid endarterectomy. METHODS AND RESULTS: Fourteen patients scheduled for carotid endarterectomy underwent preoperative carotid MRI using a custom-made phased-array coil. The plaques were excised en bloc and scanned using similar imaging parameters. MaxWA measurements from the ex vivo MRI were used as the reference standard and compared with MaxWA measurements from the corresponding in vivo MR study. Agreement between the in vivo and ex vivo measurement was analyzed using the Bland-Altman method. The paired in vivo and ex vivo MaxWA measurements strongly agreed: the mean difference (in vivo minus ex vivo) in MaxWA was 13.1+/-6.5 mm2 for T1-weighted (T1W) imaging (mean MaxWA in vivo=94.7 mm2, ex vivo=81.6 mm2) and 14.1+/-11.7 mm2 for proton density-weighted (PDW) imaging (mean MaxWA in vivo=93.4 mm2, ex vivo=79.3 mm2). Intraobserver and interobserver variability was small, with intraclass correlation coefficients ranging from 0.90 to 0.98. CONCLUSIONS: MRI is highly accurate for in vivo measurement of artery WA in atherosclerotic carotid lesions. This imaging technique has potential application monitoring lesion size in studies examining plaque progression and/or regression. PMID- 9851950 TI - Ten-year follow-up of the first megatrial testing thrombolytic therapy in patients with acute myocardial infarction: results of the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto-1 study. The GISSI Investigators. AB - BACKGROUND: We conducted a 10-year follow-up of the 11 712 patients with acute myocardial infarction randomized in the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto-1 study, the first large trial assessing thrombolytic therapy. METHODS AND RESULTS: Information on survival at 10 years was obtained for the 93% of all randomized patients through the census offices of their towns of residence. The difference in survival produced by streptokinase and sustained up to 1 year was still significant at 10 years (log-rank test, P=0.02), with the absolute benefit of 19 (95% CI 1 to 37) lives saved per 1000 patients treated. The time dependence of the extent of the benefit was confirmed, as the higher mortality rate reductions found in patients treated earlier were still present at 10 years. In the overall population, most of the benefit was obtained before hospital discharge (RR 0.81, 95% CI 0.72 to 0.90), since no difference in survival between thrombolyzed and control patients discharged alive was found at 10 years (RR 0.98, 95% CI 0.90 to 1.06). However, a slight albeit nonsignificant divergence of the survival curves of patients randomized within the first hour was observed [90 (95% CI 34 to 146) lives saved per 1000 at 10 years versus 72 (95% CI 37 to 107) lives saved at hospital discharge]. CONCLUSIONS: The benefits of a single intravenous infusion of 1.5 million units of streptokinase in prolonging survival of patients with acute myocardial infarction is sustained up to 10 years, with a still-evident trend in favor of the patients admitted earlier. PMID- 9851952 TI - Development of transplantation vasculopathy and progression of donor-transmitted atherosclerosis: comparison by serial intravascular ultrasound imaging. AB - BACKGROUND: Transplant coronary artery disease is a combination of atherosclerosis transmitted from the donor and new lesions of allograft vasculopathy. We sought to determine the morphological characteristics of allograft vasculopathy and differentiate it from donor-transmitted atherosclerosis with serial intravascular ultrasound. METHODS AND RESULTS: Intravascular ultrasound examination was performed in 93 patients at 27.2+/-15.0 and 369. 7+/-23.9 days after transplantation. The maximally and minimally diseased sites were selected in each segment as defined by Coronary Artery Surgery Study classification. For each matched site, maximal plaque thickness was measured. Lesions (maximum plaque thickness >/=0.5 mm) present at baseline examination were defined as donor lesions. On follow-up, lesions that developed at previously normal sites were defined as de novo lesions. The distribution and severity of donor and de novo lesions were similar in proximal, mid, and distal segments. The de novo lesions were less focal (43% vs 74%) and more circumferential (69% vs 45%) compared with the donor lesions, but there was significant morphological heterogeneity. Similar numbers of patients with and those without donor lesions developed de novo lesions. Moreover, progression of donor lesions was not associated with the presence or absence of de novo lesions. CONCLUSIONS: Differentiation between early allograft vasculopathy from conventional atherosclerosis by distribution and morphology of lesions alone is difficult. Serial intravascular ultrasound imaging with early baseline examination is necessary to make this distinction. This distinction is important because the progression of donor lesions and the development of de novo lesions are independent of each other. PMID- 9851953 TI - Long-term prognostic value of exercise echocardiography compared with exercise 201Tl, ECG, and clinical variables in patients evaluated for coronary artery disease. AB - BACKGROUND: The accuracy of exercise echocardiography and 201Tl single photon emission computed tomography (SPECT) is similar in the diagnosis of coronary artery disease (CAD). However, comparative data on long-term prognosis are lacking. METHODS AND RESULTS: Clinical variables and exercise, echocardiographic, and 201Tl tomographic parameters were studied in 248 patients (age, 56+/-12 years [mean+/-SD]; 189 men) who underwent simultaneous treadmill exercise 201Tl SPECT and echocardiography. Follow-up was obtained in 225 patients (91%) at a mean of 3.7+/-2.0 years. A total of 64 cardiac events occurred. With the use of stepwise logistic regression, 4 models simulating clinical stress testing scenarios were evaluated in the prediction of all cardiac events, ischemic events, and/or cardiac death. The best clinical models were exercise echocardiography with exercise ECG and exercise 201Tl SPECT with exercise ECG. Both models were comparable in the prediction of cardiac events. For the exercise echocardiography model, exercise wall motion score index and induction of ischemia were the strongest predictors of events with ORs of 2.63 per unit increment (95% CI, 1. 34 to 5.17; P=0.005) and 4.1 (95% CI, 1.32 to 12.79; P=0.015), respectively. For the model with exercise 201Tl SPECT, the strongest predictor was ischemic perfusion defect (OR, 4.93; 95% CI, 1.72 to 14.08; P=0.003). The absence of ST changes during exercise decreased the risk of events. For the prediction of ischemic events and/or cardiac death, echocardiographic and 201Tl parameters were the only predictive variables. CONCLUSIONS: In patients evaluated for CAD, exercise echocardiography and 201Tl combined with ECG variables provide comparable prognostic information and can be used interchangeably for risk stratification. PMID- 9851954 TI - Contrast magnetic resonance imaging in the assessment of myocardial viability in patients with stable coronary artery disease and left ventricular dysfunction. AB - BACKGROUND: The utility of contrast MRI for assessing myocardial viability in stable coronary artery disease (CAD) with left ventricular dysfunction is uncertain. We therefore performed cine and contrast MRI in 24 stable patients with CAD and regional contractile abnormalities and compared MRI findings with rest-redistribution 201Tl imaging and dobutamine echocardiography. METHODS AND RESULTS: Delayed MRI contrast enhancement patterns were examined from 3 to 15 minutes after injection of 0.1 mmol/kg IV gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA). Comparable MRI and 201Tl basal and midventricular short-axis images were subdivided into 6 segments. Segments judged nonviable by quantitative and qualitative assessment of 201Tl scans showed persistent, systematically greater MRI contrast signal intensity than segments judged viable (P200 times more efficient in A. tumefaciens than in Escherichia coli, as judged by the level of enzymatic activity of a lacZ-pgm fusion. PMID- 9852000 TI - The lantibiotic nisin induces transmembrane movement of a fluorescent phospholipid. AB - Nisin is a pore-forming antimicrobial peptide. The capacity of nisin to induce transmembrane movement of a fluorescent phospholipid in lipid vesicles was investigated. Unilamellar phospholipid vesicles that contained a fluorescent phospholipid (1-acyl-2-(6-[(7-nitro-2-1, 3-benzoxadiazol-4-yl)amino]caproyl)-sn glycero-3-phosphocholine) in the inner leaflet of the bilayer were used. Nisin induced movement of the fluorescent phospholipid from the inner leaflet to the outer leaflet of the membrane reached stable levels, which were dependent on the concentration of nisin added. The rate constant k of this nisin-induced transmembrane movement increased with the nisin concentration but was not dependent on temperature within the range of 5 to 30 degrees C. In contrast, the rate constant of movement of fluorescent phospholipid from vesicle to vesicle strongly depended on temperature. The data indicate that nisin transiently disturbs the phospholipid organization of the target membrane. PMID- 9852001 TI - Cytochrome bd biosynthesis in Bacillus subtilis: characterization of the cydABCD operon. AB - Under aerobic conditions Bacillus subtilis utilizes a branched electron transport chain comprising various cytochromes and terminal oxidases. At present there is evidence for three types of terminal oxidases in B. subtilis: a caa3-, an aa3-, and a bd-type oxidase. We report here the cloning of the structural genes (cydA and cydB) encoding the cytochrome bd complex. Downstream of the structural genes, cydC and cydD are located. These genes encode proteins showing similarity to bacterial ATP-binding cassette (ABC)-type transporters. Analysis of isolated cell membranes showed that inactivation of cydA or deletion of cydABCD resulted in the loss of spectral features associated with cytochrome bd. Gene disruption experiments and complementation analysis showed that the cydC and cydD gene products are required for the expression of a functional cytochrome bd complex. Disruption of the cyd genes had no apparent effect on the growth of cells in broth or defined media. The expression of the cydABCD operon was investigated by Northern blot analysis and by transcriptional and translational cyd-lacZ fusions. Northern blot analysis confirmed that cydABCD is transcribed as a polycistronic message. The operon was found to be expressed maximally under conditions of low oxygen tension. PMID- 9852002 TI - Involvement of both dockerin subdomains in assembly of the Clostridium thermocellum cellulosome. AB - Clostridium thermocellum produces an extracellular cellulase complex termed the cellulosome. It consists of a scaffolding protein, CipA, containing nine cohesin domains and a cellulose-binding domain, and at least 14 different enzymatic subunits, each containing a conserved duplicated sequence, or dockerin domain. The cohesin-dockerin interaction is responsible for the assembly of the catalytic subunits into the cellulosome structure. Each duplicated sequence of the dockerin domain contains a region bearing homology to the EF-hand calcium-binding motif. Two subdomains, each containing a putative calcium-binding motif, were constructed from the dockerin domain of CelS, a major cellulosomal catalytic subunit. These subdomains, called DS1 and DS2, were cloned by PCR and expressed in Escherichia coli. The binding of DS1 and DS2 to R3, the third cohesin domain of CipA, was analyzed by nondenaturing gel electrophoresis. A stable complex was formed only when R3 was combined with both DS1 and DS2, indicating that the two halves of the dockerin domain interact with each other and such interaction is required for effective binding of the dockerin domain to the cohesin domain. PMID- 9852003 TI - Transcriptional regulation and organization of the dcuA and dcuB genes, encoding homologous anaerobic C4-dicarboxylate transporters in Escherichia coli. AB - The dcuA and dcuB genes of Escherichia coli encode homologous proteins that appear to function as independent and mutually redundant C4-dicarboxylate transporters during anaerobiosis. The dcuA gene is 117 bp downstream of, and has the same polarity as, the aspartase gene (aspA), while dcuB is 77 bp upstream of, and has the same polarity as, the anaerobic fumarase gene (fumB). To learn more about the respective roles of the dcu genes, the environmental and regulatory factors influencing their expression were investigated by generating and analyzing single-copy dcuA- and dcuB-lacZ transcriptional fusions. The results show that dcuA is constitutively expressed whereas dcuB expression is highly regulated. The dcuB gene is strongly activated anaerobically by FNR, repressed in the presence of nitrate by NarL, and subject to cyclic AMP receptor protein (CRP) mediated catabolite repression. In addition, dcuB is strongly induced by C4 dicarboxylates, suggesting that dcuB is under the control of an uncharacterized C4-dicarboxylate-responsive gene regulator. Northern blotting confirmed that dcuA (and aspA) is expressed under both aerobic and anaerobic conditions and that dcuB (and fumB) is induced anaerobically. Major monocistronic transcripts were identified for aspA and dcuA, as well as a minor species possibly corresponding to an aspA-dcuA cotranscript. Five major transcripts were observed for dcuB and fumB: monocistronic transcripts for both fumB and dcuB; a dcuB-fumB cotranscript; and two transcripts, possibly corresponding to dcuB-fumB and fumB mRNA degradation products. Primer extension analysis revealed independent promoters for aspA, dcuA, and dcuB, but surprisingly no primer extension product could be detected for fumB. The expression of dcuB is entirely consistent with a primary role for DcuB in mediating C4-dicarboxylate transport during anaerobic fumarate respiration. The precise physiological purpose of DcuA remains unclear. PMID- 9852004 TI - Stability of the Agrobacterium tumefaciens VirB10 protein is modulated by growth temperature and periplasmic osmoadaption. AB - Export of oncogenic T-DNA from the phytopathogen Agrobacterium tumefaciens is mediated by the products of the virB operon. It has recently been reported (K. J. Fullner and E. W. Nester, J. Bacteriol. 178:1498-1504, 1996) that DNA transfer does not occur at elevated temperatures; these observations correlate well with much earlier studies on the temperature sensitivity of crown gall tumor development on plants. In testing the hypothesis that this loss of DNA movement reflects a defect in assembly or maintenance of a stable DNA transfer machinery at high temperature, we have found that steady-state levels of VirB10 are sensitive to growth temperature while levels of several other VirB proteins are considerably less affected. This temperature-dependent failure to accumulate VirB10 is exacerbated in an attachment-deficient mutant strain (chvB) which exhibits pleiotropic defects in periplasmic osmoadaption, and virulence of a chvB mutant can be partially restored by lowering the temperature at which the bacteria and the plant tissue are cocultivated. Furthermore, the stability of VirB10 is diminished in cells lacking functional VirB9, but only under conditions of low osmolarity. We propose that newly synthesized VirB10 is inherently labile in the presence of a large osmotic gradient across the inner membrane and is rapidly degraded unless it is stabilized by VirB9-dependent assembly into oligomeric complexes. The possibility that VirB10-containing complexes are not assembled properly at elevated temperatures suggests an explanation for the decades-old observation that tumor formation is exquisitely sensitive to ambient temperature. PMID- 9852005 TI - A MADS box protein consensus binding site is necessary and sufficient for activation of the opaque-phase-specific gene OP4 of Candida albicans. AB - The majority of strains of Candida albicans can switch frequently and reversibly between two or more general phenotypes, a process now considered a putative virulence factor in this species. Candida albicans WO-1 switches frequently and reversibly between a white and an opaque phase, and this phenotypic transition is accompanied by the differential expression of white-phase-specific and opaque phase-specific genes. In the opaque phase, cells differentially express the gene OP4, which encodes a putative protein 402 amino acids in length that contains a highly hydrophobic amino-terminal sequence and a carboxy-terminal sequence with a pI of 10.73. A series of deletion constructs fused to the Renilla reniformis luciferase was used to functionally characterize the OP4 promoter in order to investigate how this gene is differentially expressed in the white-opaque transition. An extremely strong 17-bp transcription activation sequence was identified between -422 and -404 bp. This sequence contained a MADS box consensus binding site, most closely related to the Mcm1 binding site of Saccharomyces cerevisiae. A number of point mutations generated in the MADS box consensus binding site as well as a complete deletion of the consensus site further demonstrated that it was essential for the activation of OP4 transcription in the opaque phase. Gel mobility shift assays with the 17-bp activation sequence identified three specific complexes which formed with both white- and opaque phase cell extracts. Competition with a putative MADS box consensus binding site from the promoter of the coordinately regulated opaque-phase-specific gene PEP1 (SAP1) and the human MADS box consensus binding site for serum response factor demonstrated that one of the three complexes formed was specific to the OP4 sequence. PMID- 9852007 TI - Topological analysis of the aerobic membrane-bound formate dehydrogenase of Escherichia coli. AB - Besides formate dehydrogenase N (FDH-N), which is involved in the major anaerobic respiratory pathway in the presence of nitrate, Escherichia coli synthesizes a second isoenzyme, called FDH-O, whose physiological role is to ensure rapid adaptation during a shift from aerobiosis to anaerobiosis. FDH-O is a membrane bound enzyme complex composed of three subunits, alpha (FdoG), beta (FdoH), and gamma (FdoI), which exhibit high sequence similarity to the equivalent polypeptides of FDH-N. The topology of these three subunits has been studied by using blaM (beta-lactamase) gene fusions. A collection of 47 different randomly generated Fdo-BlaM fusions, 4 site-specific fusions, and 3 sandwich fusions were isolated along the entire sequence of the three subunits. In contrast to previously reported predictions from sequence analysis, our data suggested that the alphabeta catalytic dimer is located in the cytoplasm, with a C-terminal anchor for beta protruding into the periplasm. As expected, the gamma subunit, which specifies cytochrome b, was shown to cross the cytoplasmic membrane four times, with the N and C termini exposed to the cytoplasm. Protease digestion studies of the 35S-labelled FDH-O heterotrimer in spheroplasts add further support to this model. Consistently, prior studies regarding the bioenergetic function of formate dehydrogenase provided evidence for a mechanism in which formate is oxidized in the cytoplasm. PMID- 9852006 TI - The Hsc66-Hsc20 chaperone system in Escherichia coli: chaperone activity and interactions with the DnaK-DnaJ-grpE system. AB - Hsc66, a stress-70 protein, and Hsc20, a J-type accessory protein, comprise a newly described Hsp70-type chaperone system in addition to DnaK-DnaJ-GrpE in Escherichia coli. Because endogenous substrates for the Hsc66-Hsc20 system have not yet been identified, we investigated chaperone-like activities of Hsc66 and Hsc20 by their ability to suppress aggregation of denatured model substrate proteins, such as rhodanese, citrate synthase, and luciferase. Hsc66 suppressed aggregation of rhodanese and citrate synthase, and ATP caused effects consistent with complex destabilization typical of other Hsp70-type chaperones. Differences in the activities of Hsc66 and DnaK, however, suggest that these chaperones have dissimilar substrate specificity profiles. Hsc20, unlike DnaJ, did not exhibit intrinsic chaperone activity and appears to function solely as a regulatory cochaperone protein for Hsc66. Possible interactions between the Hsc66-Hsc20 and DnaK-DnaJ-GrpE chaperone systems were also investigated by measuring the effects of cochaperone proteins on Hsp70 ATPase activities. The nucleotide exchange factor GrpE did not stimulate the ATPase activity of Hsc66 and thus appears to function specifically with DnaK. Cross-stimulation by the cochaperones Hsc20 and DnaJ was observed, but the requirement for supraphysiological concentrations makes it unlikely that these interactions occur significantly in vivo. Together these results suggest that Hsc66-Hsc20 and DnaK-DnaJ-GrpE comprise separate molecular chaperone systems with distinct, nonoverlapping cellular functions. PMID- 9852008 TI - The two-component regulators GacS and GacA influence accumulation of the stationary-phase sigma factor sigmaS and the stress response in Pseudomonas fluorescens Pf-5. AB - Three global regulators are known to control antibiotic production by Pseudomonas fluorescens. A two-component regulatory system comprised of the sensor kinase GacS (previously called ApdA or LemA) and GacA, a member of the FixJ family of response regulators, is required for antibiotic production. A mutation in rpoS, which encodes the stationary-phase sigma factor sigmaS, differentially affects antibiotic production and reduces the capacity of stationary-phase cells of P. fluorescens to survive exposure to oxidative stress. The gacA gene of P. fluorescens Pf-5 was isolated, and the influence of gacS and gacA on rpoS transcription, sigmaS levels, and oxidative stress response of Pf-5 was determined. We selected a gacA mutant of Pf-5 that contained a single nucleotide substitution within a predicted alpha-helical region, which is highly conserved among the FixJ family of response regulators. At the entrance to stationary phase, sigmaS content in gacS and gacA mutants of Pf-5 was less than 20% of the wild-type level. Transcription of rpoS, assessed with an rpoS-lacZ transcriptional fusion, was positively influenced by GacS and GacA, an effect that was most evident at the transition between exponential growth and stationary phase. Mutations in gacS and gacA compromised the capacity of stationary-phase cells of Pf-5 to survive exposure to oxidative stress. The results of this study provide evidence for the predominant roles of GacS and GacA in the regulatory cascade controlling stress response and antifungal metabolite production in P. fluorescens. PMID- 9852009 TI - A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus. AB - We cloned several genes encoding an Na+/H+ antiporter of Staphylococcus aureus from chromosomal DNA by using an Escherichia coli mutant, lacking all of the major Na+/H+ antiporters, as the host. E. coli cells harboring plasmids for the cloned genes were able to grow in medium containing 0.2 M NaCl (or 10 mM LiCl). Host cells without the plasmids were unable to grow under the same conditions. Na+/H+ antiport activity was detected in membrane vesicles prepared from transformants. We determined the nucleotide sequence of the cloned 7-kbp region. We found that seven open reading frames (ORFs) were necessary for antiporter function. A promoter-like sequence was found in the upstream region from the first ORF. One inverted repeat followed by a T-cluster, which may function as a terminator, was found in the downstream region from the seventh ORF. Neither terminator-like nor promoter-like sequences were found between the ORFs. Thus, it seems that the seven ORFs comprise an operon and that the Na+/H+ antiporter consists of seven kinds of subunits, suggesting that this is a novel type of multisubunit Na+/H+ antiporter. Hydropathy analysis of the deduced amino acid sequences of the seven ORFs suggested that all of the proteins are hydrophobic. As a result of a homology search, we found that components of the respiratory chain showed sequence similarity with putative subunits of the Na+/H+ antiporter. We observed a large Na+ extrusion activity, driven by respiration in E. coli cells harboring the plasmid carrying the genes. The Na+ extrusion was sensitive to an H+ conductor, supporting the idea that the system is not a respiratory Na+ pump but an Na+/H+ antiporter. Introduction of the plasmid into E. coli mutant cells, which were unable to grow under alkaline conditions, enabled the cells to grow under such conditions. PMID- 9852010 TI - Expression of the Bacillus subtilis acsA gene: position and sequence context affect cre-mediated carbon catabolite repression. AB - In Bacillus subtilis, carbon catabolite repression (CCR) of many genes is mediated at cis-acting carbon repression elements (cre) by the catabolite repressor protein CcpA. Mutations in transcription-repair coupling factor (mfd) partially relieve CCR at cre sites located downstream of transcriptional start sites by abolishing the Mfd-mediated displacement of RNA polymerase stalled at cre sites which act as transcriptional roadblocks. Although the acsA cre is centered 44.5 bp downstream of the acsA transcriptional start site, CCR of acsA expression is not affected by an mfd mutation. When the acsA cre is centered 161.5 bp downstream of the transcriptional start site for the unregulated tms promoter, CCR is partially relieved by the mfd mutation. Since CCR mediated at an acsA cre centered 44.5 bp downstream of the tms start site is not affected by the mfd mutation, the inability of Mfd to modulate CCR of acsA expression most likely results from the location of the acsA cre. Higher levels of CCR were found to occur at cre sites flanked by A+T-rich sequences than at cre sites bordered by G and C nucleotides. This suggests that nucleotides adjacent to the proposed 14-bp cre consensus sequence participate in the formation of the CcpA catabolite repression complex at cre sites. Examination of CCR of acsA expression revealed that this regulation required the Crh and seryl-phosphorylated form of the HPr proteins but not glucose kinase. PMID- 9852011 TI - Functional similarities between the Listeria monocytogenes virulence regulator PrfA and cyclic AMP receptor protein: the PrfA* (Gly145Ser) mutation increases binding affinity for target DNA. AB - Most Listeria monocytogenes virulence genes are positively regulated by the PrfA protein, a transcription factor sharing sequence similarities with cyclic AMP (cAMP) receptor protein (CRP). Its coding gene, prfA, is regulated by PrfA itself via an autoregulatory loop mediated by the upstream PrfA-dependent plcA promoter. We have recently characterized prfA* mutants from L. monocytogenes which, as a result of a single amino acid substitution in PrfA, Gly145Ser, constitutively overexpress prfA and the genes of the PrfA virulence regulon. Here, we show that about 10 times more PrfA protein is produced in a prfA* strain than in the wild type. Thus, the phenotype of prfA* mutants is presumably due to the synthesis of a PrfA protein with higher promoter-activating activity (PrfA*), which keeps its intracellular levels constantly elevated by positive feedback. We investigated the interaction of PrfA and PrfA* (Gly145Ser) with target DNA. Gel retardation assays performed with a DNA fragment carrying the PrfA binding site of the plcA promoter demonstrated that the PrfA* mutant form is much more efficient than wild type PrfA at forming specific DNA-protein complexes. In footprinting experiments, the two purified PrfA forms interacted with the same nucleotides at the target site, although the minimum amount required for protection was 6 to 7 times lower with PrfA*. These results show that the primary functional consequence of the Gly145Ser mutation is an increase in the affinity of PrfA for its target sequence. Interestingly, similar mutations at the equivalent position in CRP result in a transcriptionally active, CRP* mutant form which binds with high affinity to target DNA in the absence of the activating cofactor, cAMP. Our observations suggest that the structural similarities between PrfA and CRP are also functionally relevant and support a model in which the PrfA protein, like CRP, shifts from transcriptionally inactive to active conformations by interaction with a cofactor. PMID- 9852012 TI - Physiological role for the GlnK protein of enteric bacteria: relief of NifL inhibition under nitrogen-limiting conditions. AB - In Klebsiella pneumoniae, NifA-dependent transcription of nitrogen fixation (nif) genes is inhibited by a flavoprotein, NifL, in the presence of molecular oxygen and/or combined nitrogen. We recently demonstrated that the general nitrogen regulator NtrC is required to relieve NifL inhibition under nitrogen (N)-limiting conditions. We provide evidence that the sole basis for the NtrC requirement is its role as an activator of transcription for glnK, which encodes a PII-like allosteric effector. Relief of NifL inhibition is a unique physiological function for GlnK in that the structurally related GlnB protein of enteric bacteria apparently a paralogue of GlnK-cannot substitute. Unexpectedly, although covalent modification of GlnK by uridylylation normally occurs under N-limiting conditions, several lines of evidence indicate that uridylylation is not required for relief of NifL inhibition. When GlnK was synthesized constitutively from non NtrC-dependent promoters, it was able to relieve NifL inhibition in the absence of uridylyltransferase, the product of the glnD gene, and under N excess conditions. Moreover, an altered form of GlnK, GlnKY51N, which cannot be uridylylated due to the absence of the requisite tyrosine, was still able to relieve NifL inhibition. PMID- 9852013 TI - The modified beta-ketoadipate pathway in Rhodococcus rhodochrous N75: enzymology of 3-methylmuconolactone metabolism. AB - Rhodococcus rhodochrous N75 is able to metabolize 4-methylcatechol via a modified beta-ketoadipate pathway. This organism has been shown to activate 3 methylmuconolactone by the addition of coenzyme A (CoA) prior to hydrolysis of the butenolide ring. A lactone-CoA synthetase is induced by growth of R. rhodochrous N75 on p-toluate as a sole source of carbon. The enzyme has been purified 221-fold by ammonium sulfate fractionation, hydrophobic chromatography, gel filtration, and anion-exchange chromatography. The enzyme, termed 3 methylmuconolactone-CoA synthetase, has a pH optimum of 8.0, a native Mr of 128,000, and a subunit Mr of 62,000, suggesting that the enzyme is homodimeric. The enzyme is very specific for its 3-methylmuconolactone substrate and displays little or no activity with other monoene and diene lactone analogues. Equimolar amounts of these lactone analogues brought about less than 30% (most brought about less than 15%) inhibition of the CoA synthetase reaction with its natural substrate. PMID- 9852014 TI - The yvyD gene of Bacillus subtilis is under dual control of sigmaB and sigmaH. AB - During a search by computer-aided inspection of two-dimensional (2D) protein gels for sigmaB-dependent general stress proteins exhibiting atypical induction profiles, a protein initially called Hst23 was identified as a product of the yvyD gene of Bacillus subtilis. In addition to the typical sigmaB-dependent, stress- and starvation-inducible pattern, yvyD is also induced in response to amino acid depletion. By primer extension of RNA isolated from the wild-type strain and appropriate mutants carrying mutations in the sigB and/or spo0H gene, two promoters were mapped upstream of the yvyD gene. The sigmaB-dependent promoter drives expression of yvyD under stress conditions and after glucose starvation, whereas a sigmaH-dependent promoter is responsible for yvyD transcription following amino acid limitation. Analysis of Northern blots revealed that yvyD is transcribed monocistronically and confirmed the conclusions drawn from the primer extension experiments. The analysis of the protein synthesis pattern in amino acid-starved wild-type and relA mutant cells showed that the YvyD protein is not synthesized in the relA mutant background. It was concluded that the stringent response plays a role in the activation of sigmaH. The yvyD gene product is homologous to a protein which might modify the activity of sigma54 in gram-negative bacteria. The expression of a sigmaL-dependent (sigmaL is the equivalent of sigma54 in B. subtilis) levD-lacZ fusion is upregulated twofold in a yvyD mutant. This indicates that the yvyD gene product, being a member of both the sigmaB and sigmaH regulons, might negatively regulate the activity of the sigmaL regulon. We conclude that (i) systematic, computer aided analysis of 2D protein gels is appropriate for the identification of genes regulated by multiple transcription factors and that (ii) YvyD might form a junction between the sigmaB and sigmaH regulons on one side and the sigmaL regulon on the other. PMID- 9852015 TI - The first gene of the Bacillus subtilis clpC operon, ctsR, encodes a negative regulator of its own operon and other class III heat shock genes. AB - The Bacillus subtilis clpC operon is regulated by two stress induction pathways relying on either sigmaB or a class III stress induction mechanism acting at a sigmaA-like promoter. When the clpC operon was placed under the control of the isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible Pspac promoter, dramatic repression of the natural clpC promoters fused to a lacZ reporter gene was noticed after IPTG induction. This result strongly indicated negative regulation of the clpC operon by one of its gene products. Indeed, the negative regulator could be identified which is encoded by the first gene of the clpC operon, ctsR, containing a predicted helix-turn-helix DNA-binding motif. Deletion of ctsR abolished the negative regulation and resulted in high expression of both the clpC operon and the clpP gene under nonstressed conditions. Nevertheless, a further increase in clpC and clpP mRNA levels was observed after heat shock, even in the absence of sigmaB, suggesting a second induction mechanism at the vegetative promoter. Two-dimensional gel analysis and mRNA studies showed that the expression of other class III stress genes was at least partially influenced by the ctsR deletion. Studies with different clpC promoter fragments either fused to the reporter gene bgaB or used in gel mobility shift experiments with the purified CtsR protein revealed a possible target region where the repressor seemed to bind in vivo and in vitro. Our data demonstrate that the CtsR protein acts as a global repressor of the clpC operon, as well as other class III heat shock genes, by preventing unstressed transcription from either the sigmaB- or sigmaA-dependent promoter and might be inactivated or dissociate under inducing stress conditions. PMID- 9852017 TI - Genome size, complexity, and ploidy of the pathogenic fungus Histoplasma capsulatum. AB - The genome size, complexity, and ploidy of the dimorphic pathogenic fungus Histoplasma capsulatum was determined by using DNA renaturation kinetics, genomic reconstruction, and flow cytometry. Nuclear DNA was isolated from two strains, G186AS and Downs, and analyzed by renaturation kinetics and genomic reconstruction with three putative single-copy genes (calmodulin, alpha-tubulin, and beta-tubulin). G186AS was found to have a genome of approximately 2.3 x 10(7) bp with less than 0.5% repetitive sequences. The Downs strain, however, was found to have a genome approximately 40% larger with more than 16 times more repetitive DNA. The Downs genome was determined to be 3.2 x 10(7) bp with approximately 8% repetitive DNA. To determine ploidy, the DNA mass per cell measured by flow cytometry was compared with the 1n genome estimate to yield a DNA index (DNA per cell/1n genome size). Strain G186AS was found to have a DNA index of 0.96, and Downs had a DNA index of 0.94, indicating that both strains are haploid. Genomic reconstruction and Southern blot data obtained with alpha- and beta-tubulin probes indicated that some genetic duplication has occurred in the Downs strain, which may be aneuploid or partially diploid. PMID- 9852016 TI - Selectivity of ferric enterobactin binding and cooperativity of transport in gram negative bacteria. AB - The ligand-gated outer membrane porin FepA serves Escherichia coli as the receptor for the siderophore ferric enterobactin. We characterized the ability of seven analogs of enterobactin to supply iron via FepA by quantitatively measuring the binding and transport of their 59Fe complexes. The experiments refuted the idea that chirality of the iron complex affects its recognition by FepA and demonstrated the necessity of an unsubstituted catecholate coordination center for binding to the outer membrane protein. Among the compounds we tested, only ferric enantioenterobactin, the synthetic, left-handed isomer of natural enterobactin, and ferric TRENCAM, which substitutes a tertiary amine for the macrocyclic lactone ring of ferric enterobactin but maintains an unsubstituted catecholate iron complex, were recognized by FepA (Kd approximately 20 nM). Ferric complexes of other analogs (TRENCAM-3,2-HOPO; TREN-Me-3,2-HOPO; MeMEEtTAM; MeME-Me-3,2-HOPO; K3MECAMS; agrobactin A) with alterations to the chelating groups and different net charge on the iron center neither adsorbed to nor transported through FepA. We also compared the binding and uptake of ferric enterobactin by homologs of FepA from Bordetella bronchisepticus, Pseudomonas aeruginosa, and Salmonella typhimurium in the native organisms and as plasmid mediated clones expressed in E. coli. All the transport proteins bound ferric enterobactin with high affinity (Kd /=50 pmol/min/10(9) cells) in their own particular membrane environments. However, the FepA and IroN proteins of S. typhimurium failed to efficiently function in E. coli. For E. coli, S. typhimurium, and P. aeruginosa, the rate of ferric enterobactin uptake was a sigmoidal function of its concentration, indicating a cooperative transport reaction involving multiple interacting binding sites on FepA. PMID- 9852018 TI - New small, acid-soluble proteins unique to spores of Bacillus subtilis: identification of the coding genes and regulation and function of two of these genes. AB - Eleven small, acid-soluble proteins (SASP) which are present in spores but not in growing cells of Bacillus subtilis were identified by sequence analysis of proteins separated by acrylamide gel electrophoresis of acid extracts from spores which lack the three major SASP (alpha, beta, and gamma). Six of these proteins are encoded by open reading frames identified previously or by analysis of the complete sequence of the B. subtilis genome, including two minor alpha/beta-type SASP (SspC and SspD) and a putative spore coat protein (CotK). Five proteins are encoded by short open reading frames that were not identified as coding regions in the analysis of the complete B. subtilis genomic sequence. Studies of the regulation of two of the latter genes, termed sspG and sspJ, showed that both are expressed only in sporulation. The sspG gene is transcribed in the mother cell compartment by RNA polymerase with the mother cell-specific sigma factor for RNA polymerase, sigmaK, and is cotranscribed with a downstream gene, yurS; sspG transcription also requires the DNA binding protein GerE. In contrast, sspJ is transcribed in the forespore compartment by RNA polymerase with the forespore specific sigmaG and appears to give a monocistronic transcript. A mutation eliminating SspG had no effect on sporulation or spore properties, while loss of SspJ caused a slight decrease in the rate of spore outgrowth in an otherwise wild type background. PMID- 9852019 TI - Comparison in vitro of a high- and a low-abundance chemoreceptor of Escherichia coli: similar kinase activation but different methyl-accepting activities. AB - In Escherichia coli, high-abundance chemoreceptors are present in cellular amounts approximately 10-fold greater than low-abundance chemoreceptors. Cells containing only low-abundance receptors exhibit abnormally low tumble frequencies and do not migrate effectively in spatial gradients. These defects reflect an inherent activity difference between the two receptor classes. We used in vitro assays to investigate this difference. The low-abundance receptor Trg mediated an approximately 100-fold activation of the kinase CheA, only twofold less than activation by the high-abundance receptor Tar. In contrast, Trg was less than 1/20 as active as Tar for in vitro methylation. As observed for high-abundance receptors, kinase activation by Trg varied with the extend of modification at methyl-accepting sites; low methylation corresponded to low kinase activation. Thus, in Trg-only cells, low receptor methylation would result in low kinase activation, correspondingly low content of phospho-CheY, and a decreased dynamic range over which attractant binding could modulate kinase activity. These features could account for the low tumble frequency and inefficient taxis exhibited by Trg-only cells. Thus, the crucial functional difference between the receptor classes is likely to be methyl-accepting activity. We investigated the structural basis for this functional difference by introducing onto the carboxy terminus of Trg a CheR-binding pentapeptide, usually found only at the carboxy termini of high-abundance receptors. This addition enhanced the in vitro methyl accepting activity of Trg 10-fold. PMID- 9852020 TI - Coupled changes in translation and transcription during cobalamin-dependent regulation of btuB expression in Escherichia coli. AB - The level of the vitamin B12 transport protein BtuB in the outer membrane of Escherichia coli is strongly reduced by growth in the presence of cobalamins. Previous analyses of regulatory mutants and of btuB-lacZ fusions indicated that the primary site of btuB gene regulation was at the translational level, and this required sequences throughout the 240-nucleotide (nt) leader region. Cobalamin dependent regulation of transcriptional fusions was of a lesser magnitude but required, in addition to the leader, sequences within the first 100 nt of the coding sequence, termed the translated regulatory region (TRR). To analyze the process of transcription-level regulation of btuB in E. coli, the levels and metabolism of btuB RNA were analyzed by S1 nuclease protection assays, and mutations that alter the coupling of translational and transcriptional control were analyzed. Expression of transcriptional fusions was found to correlate with changes in the level of intact btuB RNA and was related to changes in the metabolic stability of the normally long-lived RNA. Mutational analysis showed that the btuB start codon and a hairpin structure that can sequester the Shine Dalgarno sequence are necessary for cobalamin-dependent regulation and that translation of the TRR is necessary for extended RNA stability and for expression of the transcriptional fusion. The absence of regulation at the stage of transcription initiation was confirmed by the findings that several truncated btuB RNA fragments were expressed in a constitutive manner and that the normal regulatory response occurred even when the btuB promoter and upstream sequences were replaced by the heterologous bla and lac promoters. Transcription driven by phage T7 RNA polymerase was not regulated by cobalamins, although some regulation at the translational level was retained. Cobalamin-dependent changes in RNA structure were suggested from the RNase III-dependent production of a transcript fragment that is made only in the presence of cobalamin and is independent of the regulatory outcome. These results indicate that the primary control of btuB expression by cobalamin occurs at the level of translation initiation, which directly affects the level and stability of btuB RNA in a process that requires the presence of the intact translated regulatory region. PMID- 9852021 TI - Role of the gerI operon of Bacillus cereus 569 in the response of spores to germinants. AB - Bacillus cereus 569 (ATCC 10876) germinates in response to inosine or to L alanine, but the most rapid germination response is elicited by a combination of these germinants. Mutants defective in their germination response to either inosine or to L-alanine were isolated after Tn917-LTV1 mutagenesis and enrichment procedures; one class of mutant could not germinate in response to inosine as a sole germinant but still germinated in response to L-alanine, although at a reduced rate; another mutant germinated normally in response to inosine but was slowed in its germination response to L-alanine. These mutants demonstrated that at least two signal response pathways are involved in the triggering of germination. Stimulation of germination in L-alanine by limiting concentrations of inosine and stimulation of germination in inosine by low concentrations of L alanine were still detectable in these mutants, suggesting that such stimulation is not dependent on complete functionality of both these germination loci. Two transposon insertions that affected inosine germination were found to be located 2.2 kb apart on the chromosome. This region was cloned and sequenced, revealing an operon of three open reading frames homologous to those in the gerA and related operons of Bacillus subtilis. The individual genes of this gerI operon have been named gerIA, gerIB, and gerIC. The GerIA protein is predicted to possess an unusually long, charged, N-terminal domain containing nine tandem copies of a 13-amino-acid glutamine- and serine-rich sequence. PMID- 9852022 TI - Disruption of PMR1, encoding a Ca2+-ATPase homolog in Yarrowia lipolytica, affects secretion and processing of homologous and heterologous proteins. AB - The Yarrowia lipolytica PMR1 gene (YlPMR1) is a Saccharomyces cerevisiae PMR1 homolog which encodes a putative secretory pathway Ca2+-ATPase. In this study, we investigated the effects of a YlPMR1 disruption on the processing and secretion of native and foreign proteins in Y. lipolytica and found variable responses by the YlPMR1-disrupted mutant depending on the protein. The secretion of 32-kDa mature alkaline extracellular protease (AEP) was dramatically decreased, and incompletely processed precursors were observed in the YlPMR1-disrupted mutant. A 36- and a 52-kDa premature AEP were secreted, and an intracellular 52-kDa premature AEP was also detected. The acid extracellular protease activity of the YlPMR1-disrupted mutant was increased by 60% compared to that of the wild-type strain. The inhibitory effect of mutations in secretory pathway Ca2+-ATPase genes on the secretion of rice alpha-amylase was also observed in the Y. lipolytica and S. cerevisiae PMR1-disrupted mutants. Unlike rice alpha-amylase, the secretion of Trichoderma reesei endoglucanase I (EGI) was not influenced by the YlPMR1 disruption. However, the secreted EGI from the YlPMR1-disrupted mutant had different characteristics than that of the control. While wild-type cells secreted the hyperglycosylated form of EGI, hyperglycosylation was completely absent in the YlPMR1-disrupted mutant. Our results indicate that the effects of the YlPMR1 disruption as manifested by the phenotypic response depend on the characteristics of the reporter protein in the recombinant yeast strain evaluated. PMID- 9852023 TI - Superimposition of tyrR protein-mediated regulation on osmoresponsive transcription of Escherichia coli proU in vivo. AB - Osmotic regulation of proU expression in the enterobacteria is achieved, at least in part, by a repression mechanism involving the histone-like nucleoid protein H NS. By the creation of binding sites for the TyrR regulator protein in the vicinity of the sigma70-controlled promoter of proU in Escherichia coli, we were able to demonstrate a superposed TyrR-mediated activation by L-phenylalanine (Phe), as well as repression by L-tyrosine, of proU expression in vivo. Based on the facts that pronounced activation in the presence of Phe was observed even at a low osmolarity and that the affinity of binding of TyrR to its cognate sites on DNA is not affected by Phe, we argue that H-NS-mediated repression of proU at a low osmolarity may not involve a classical silencing mechanism. Our data also suggest the involvement of recruited RNA polymerase in the mechanism of antirepression in E. coli. PMID- 9852024 TI - Cloning and expression of the gene for the Na+-coupled serine transporter from Escherichia coli and characteristics of the transporter. AB - We cloned a gene (sstT) for the Na+/serine symporter from the chromosome of Escherichia coli by using a low-copy-number vector and sequenced it. According to the deduced amino acid sequence, the transporter (SstT) consists of 414 amino acid residues. Hydropathy analysis suggested that the SstT protein possesses 9, instead of 12, hydrophobic domains. PMID- 9852025 TI - Insertion mutagenesis of the ferric pyoverdine receptor FpvA of Pseudomonas aeruginosa: identification of permissive sites and a region important for ligand binding. AB - Insertion of an 18-amino-acid-encoding sequence within the fpvA gene identified permissive sites at residues Y350, A402, R451, R521, and R558, consistent with these residues occurring in extramembranous loop regions of the protein. Insertions at R451, R521, and R558 did not adversely affect receptor function, although insertions at Y350 and A402 compromised ferric pyoverdine binding and uptake. The latter region likely contributes to or interacts with the ligand binding site. PMID- 9852026 TI - The aadA gene of plasmid R100 confers resistance to spectinomycin and streptomycin in Myxococcus xanthus. AB - Plasmids with the aadA gene from plasmid R100, which confers resistance to the aminoglycosides spectinomycin and streptomycin in Escherchia coli, can be introduced into wild-type Myxococcus xanthus, strain DK1622, by electroporation. Recombinant M. xanthus strains with integrated plasmids carrying the aadA gene acquire resistance to high levels of these antibiotics. Selection for aadA in M. xanthus can be carried out independently of, or simultaneously with, selection for resistance to kanamycin. The kinds and frequencies of recombination events observed between integrative plasmids with aadA and the M. xanthus chromosome are similar to those observed after the transformation of yeast. Cleavage of integrative plasmid DNA at a site adjacent to a region of homology between the plasmid and the M. xanthus genome favors the targeted disruption of M. xanthus genes by allele replacement. PMID- 9852027 TI - Nitrite-specific active transport system of the cyanobacterium Synechococcus sp. strain PCC 7942. AB - Studies on the nitrite uptake capability of a mutant of Synechococcus sp. strain PCC 7942 lacking the ATP-binding cassette-type nitrate-nitrite-bispecific transporter revealed the occurrence of a nitrite-specific active transport system with an apparent Km (NO2-) of about 20 microM. Similar to the nitrate-nitrite bispecific transporter, the nitrite-specific transporter was reversibly inhibited by ammonium in the medium. PMID- 9852028 TI - A novel serine/threonine protein kinase homologue of Pseudomonas aeruginosa is specifically inducible within the host infection site and is required for full virulence in neutropenic mice. AB - A genetic locus of Pseudomonas aeruginosa was identified that is highly and specifically inducible during infection of neutropenic mice. This locus, ppkA, encodes a protein that is highly homologous to eukaryote-type serine/threonine protein kinases. A ppkA null mutant strain shows reduced virulence in neutropenic mice compared to the wild type. Overexpression of the PpkA protein greatly inhibited the growth of Escherichia coli or P. aeruginosa. However, a single amino acid change at the catalytic site of the kinase domain eliminated the toxic effect of PpkA on bacterial cells, suggesting that the kinase domain of PpkA is functional within bacterial cells. PMID- 9852029 TI - Active efflux of organic solvents by Pseudomonas putida S12 is induced by solvents. AB - Induction of the membrane-associated organic solvent efflux system SrpABC of Pseudomonas putida S12 was examined by cloning a 312-bp DNA fragment, containing the srp promoter, in the broad-host-range reporter vector pKRZ-1. Compounds that are capable of inducing expression of the srpABC genes include aromatic and aliphatic solvents and alcohols. General stress conditions such as pH, temperature, NaCl, or the presence of organic acids did not induce srp transcription. Although the solvent efflux pump in P. putida S12 is a member of the resistance-nodulation-cell division family of transporters, the srpABC genes were not induced by antibiotics or heavy metals. PMID- 9852030 TI - A mutant of Mycobacterium smegmatis defective in dipeptide transport. AB - A mutant of Mycobacterium smegmatis unable to use the dipeptide carnosine (beta alanyl-L-histidine) as a sole carbon or nitrogen source was isolated. Carnosinase activity and the ability to grow on beta-Ala and/or L-His were similar in the mutant and the wild type. However, the mutant showed significant impairment in the uptake of carnosine. This study is the first description of a peptide utilization mutant of a mycobacterium. PMID- 9852031 TI - Sucrose-phosphate synthase from Synechocystis sp. strain PCC 6803: identification of the spsA gene and characterization of the enzyme expressed in Escherichia coli. AB - The first identification and characterization of a prokaryotic gene (spsA) encoding sucrose-phosphate synthase (SPS) is reported for Synechocystis sp. strain PCC 6803, a unicellular non-nitrogen-fixing cyanobacterium. Comparisons of the deduced amino acid sequence and some relevant biochemical properties of the enzyme with those of plant SPSs revealed important differences in the N-terminal and UDP-glucose binding site regions, substrate specificities, molecular masses, subunit compositions, and regulatory properties. PMID- 9852032 TI - Identification of two binding domains, one for peptidoglycan and another for a secondary cell wall polymer, on the N-terminal part of the S-layer protein SbsB from Bacillus stearothermophilus PV72/p2. AB - First studies on the structure-function relationship of the S-layer protein from B. stearothermophilus PV72/p2 revealed the coexistence of two binding domains on its N-terminal part, one for peptidoglycan and another for a secondary cell wall polymer (SCWP). The peptidoglycan binding domain is located between amino acids 1 to 138 of the mature S-layer protein comprising a typical S-layer homologous domain. The SCWP binding domain lies between amino acids 240 to 331 and possesses a high serine plus glycine content. PMID- 9852033 TI - The fur-regulated gene encoding the alternative sigma factor PvdS is required for iron-dependent expression of the LysR-type regulator ptxR in Pseudomonas aeruginosa. AB - We previously identified a novel regulator of the exotoxin A gene (toxA) in Pseudomonas aeruginosa, PtxR, that belongs to the LysR family of prokaryotic regulatory proteins. Preliminary data also suggest that PtxR affects the expression of siderophores in P. aeruginosa. Because toxA expression and siderophore production in this organism are coordinately regulated by the ferric uptake regulator (Fur) and the Fur-regulated alternative sigma factor PvdS, regulation of ptxR itself in the context of these regulators was examined. RNase protection analyses of ptxR transcription revealed that there are two independent transcription initiation sites (T1 and T2). While transcription from the promoter of T1 is constitutive throughout the growth cycle of PAO1, transcription from the second promoter (P2) is negatively affected by iron. Transcription from the P2 promoter is constitutive in a fur mutant under microaerobic conditions but still iron regulated during aerobic growth. High concentrations (>100 nM) of the ferric uptake regulatory protein (Fur) failed to bind to either of the promoter regions of ptxR in either gel mobility shift assays or DNase I footprint experiments. These results indicate that Fur indirectly regulates the iron-dependent expression of ptxR. Iron-regulated transcription of ptxR from the P2 promoter, but not constitutive expression from the P1 promoter, was dependent on the Fur regulated alternative sigma factor gene pvdS, even under aerobic conditions. Consequently, there are two levels of iron-regulated expression of ptxR. The iron regulated expression of ptxR under microaerobic conditions from the P2 promoter of ptxR is mediated indirectly by Fur through the iron-regulated expression of pvdS. In contrast, pvdS-mediated iron regulation of ptxR under aerobic conditions is Fur independent. PMID- 9852034 TI - Identification and characterization of the Erwinia amylovora rpoS gene: RpoS is not involved in induction of fireblight disease symptoms. AB - The Erwinia amylovora rpoS gene, encoding the alternative sigma factor RpoS, has been cloned and characterized. Though highly sensitive to a number of environmental stresses, an E. amylovora rpoS mutant was not compromised in its ability to grow or cause disease symptoms within apple seedlings or in an overwintering model. PMID- 9852035 TI - Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene. AB - Recently, the growth hormone secretagogue receptor (GHS-R) cDNA has been isolated from the pituitary and hypothalamus. To evaluate the regulation of human (h) GHS R gene expression, we cloned the hGHS-R gene containing the 5'-flanking region of 0.6-2.9 kilobase pairs. Analysis of the hGHS-R transcripts with 5'-rapid amplification of cDNA ends suggested that the putative transcription initiation site was approximately -453 base pairs upstream of the translation initiation site (+1). There is no typical TATA, CAAT, or GC box but an initiator-like sequence and putative binding sites for several transcription factors around the putative transcription start site. The 5'-flanking region inserted into a luciferase reporter vector had promoter activity in GH3 cells but had activity indistinguishable from background in HeLa or EP1 cells. The hGHS-R promoter activity in GH3 cells increased by deletion of nucleotides from -1224 to -734, whereas it was decreased by further deletion from -734 to -608. Knowledge of the promoter region of the hGHS-R gene will facilitate elucidation of its transcriptional control. PMID- 9852036 TI - A new protein conjugation system in human. The counterpart of the yeast Apg12p conjugation system essential for autophagy. AB - Autophagy is an intracellular process for bulk degradation of cytoplasmic components. We recently found a protein conjugation system essential for autophagy in the yeast, Saccharomyces cerevisiae. The C-terminal glycine of a novel modifier protein, Apg12p, is conjugated to a lysine residue of Apg5p via an isopeptide bond. This conjugation reaction is mediated by Apg7p, a ubiquitin activating enzyme (E1)-like enzyme, and Apg10p, suggesting that it is a ubiquitination-like system (Mizushima, N., Noda, T., Yoshimori, T., Tanaka, Y., Ishii, T., George, M. D., Klionsky, D. J., Ohsumi, M. , and Ohsumi, Y. (1998) Nature 395, 395-398). Although autophagy is a ubiquitous process in eukaryotic cells, no molecule involved in autophagy has yet been identified in higher eukaryotes. We reasoned that this conjugation system could be conserved. Here we report cloning and characterization of the human homologue of Apg12 (hApg12). It is a 140-amino acid protein and possesses 27% identity and 48% similarity with the yeast Apg12p, but no apparent homology to ubiquitin. Northern blot analysis showed that its expression was ubiquitous in human tissues. We found that it was covalently attached to another protein. This target protein was identified to be the human Apg5 homologue (hApg5). Mutagenic analyses suggested that this conjugation was formed via an isopeptide bond between the C-terminal glycine of hApg12 and Lys-130 of hApg5. These findings indicate that the Apg12 system is well conserved and may function in autophagy also in human cells. PMID- 9852037 TI - Protein-tyrosine phosphatase Shp-1 is a negative regulator of IL-4- and IL-13 dependent signal transduction. AB - Binding of interleukin (IL)-4 to its transmembrane receptor results in the Jak mediated tyrosine phosphorylation of a number of protein components of the IL-4 signaling cascade, including Jak1, Jak2, Jak3, Tyk2, IL-4Ralpha, IRS-1, IRS-2, and Stat6 in appropriate cell types. However, the protein-tyrosine phosphatases (PTPs) that dephosphorylate these proteins and terminate signaling remained unidentified. We have noted that IL-4-dependent activation of Stat6 is sustained longer in fibroblasts than in lymphoid cells. Because Shp-1, an SH2 domain containing PTP, is expressed primarily in hematopoietic cells, we examined whether Shp-1 activity could regulate IL-4-dependent cell signaling. Expression of an Shp-1 transgene in NIH 3T3 cells markedly reduces both IL-4-dependent Stat6 activation and Stat6-mediated transcription of IL-4-responsive genes. In accord with this, IL-4 treatment of bone marrow-derived macrophages from viable motheaten mice that express substantially reduced levels of Shp-1 activity show remarkably enhanced activation of Stat6. In addition, Stat6 activation by IL-4 is significantly enhanced in pre-B cells derived from motheaten (Shp-1 null mutant) mice compared with normal pre-B cells derived from control animals. These data clearly implicate Shp-1 in the negative regulation of the IL-4/IL-13-activated Jak-Stat pathway. PMID- 9852038 TI - The death effector domain of PEA-15 is involved in its regulation of integrin activation. AB - Increased integrin ligand binding affinity (activation) is triggered by intracellular signaling events. A Ras-initiated mitogen-activated protein kinase pathway suppresses integrin activation in fibroblasts. We used expression cloning to isolate cDNAs that prevent Ras suppression of integrin activation. Here, we report that PEA-15, a small death effector domain (DED)-containing protein, blocks Ras suppression. PEA-15 does not block the capacity of Ras to activate the ERK mitogen-activated protein kinase pathway. Instead, it inhibits suppression via a pathway blocked by a dominant-negative form of the distinct small GTPase, R Ras. Heretofore, all known DEDs functioned in the regulation of apoptosis. In contrast, the DED of PEA-15 is essential for its capacity to reverse suppression of integrin activation. Thus, certain DED-containing proteins can regulate integrin activation as opposed to apoptotic protease cascades. PMID- 9852039 TI - Interferon-gamma selectively induces Rab5a synthesis and processing in mononuclear cells. AB - Macrophage activation by interferon (IFN)-gamma is characterized by enhanced phagocytosis and killing of internalized pathogens. We studied the effects of IFN gamma on Rab5a, a GTPase involved in both endocytosis and phagocytosis. IFN-gamma induced the synthesis of Rab5a in mononuclear cells as detected by immunoprecipitation and by Western blotting. Rab5a messenger RNA levels were also increased. Elevated protein expression was detected as early as 6 h following IFN gamma and was maximal at 24 h. Following IFN-gamma, membrane association of Rab5a:GTP was substantially increased. Rab5b and Rab5c, as well as Rab7 and Rab11, Rab GTPases localized in the endosomal-lysosomal pathway, were unaffected by IFN-gamma. Moreover, Rab5a expression in non-macrophages was unaltered by IFN gamma. Rab5a is a prenylated protein, and newly synthesized Rab5a was rapidly processed following IFN-gamma. However, elevated geranylgeranylation was not Rab5a-specific since all the Rab5 isoforms were more rapidly prenylated in vitro using cytosol from IFN-gamma-treated cells. Last, guanine nucleotide exchange on Rab5a was elevated about 3-fold in the presence of cytosol from IFN-gamma-treated cells. The selective effect of IFN-gamma on Rab5a, synthesis, processing, and nucleotide exchange suggests that Rab isoforms have closely associated but not identical functions and that selective enhancement of membrane trafficking may play a key role in intracellular killing. PMID- 9852040 TI - The changes in adenine nucleotides measured in glucose-stimulated rodent islets occur in beta cells but not in alpha cells and are also observed in human islets. AB - Glucose metabolism by pancreatic beta and alpha cells is essential for stimulation of insulin secretion and inhibition of glucagon secretion. Studies using rodent islets have suggested that the ATP/ADP ratio serves as second messenger in beta cells. This study compared the effects of glucose on glucose oxidation ([U-14C]glucose) and adenine nucleotides (luminometric method) in purified rat alpha and beta cells. The rate of glucose oxidation at 1 mM glucose was higher in beta than alpha cells (4.5-fold, i.e. approximately 2-fold after normalization for cell size). It was more strongly stimulated by 10 mM glucose in beta cells (9-fold) than in alpha cells (5-fold). At 1 mM glucose, ATP levels were similar in both cell types, which corresponds to an approximately 2-fold higher concentration in alpha cells ( approximately 6.5 mM) than in beta cells ( approximately 3 mM). In beta cells, glucose dose-dependently increased ATP and decreased ADP levels, causing a rise in the ATP/ADP ratio from 2.4 to 11.6 at 1 and 10 mM, respectively. In alpha cells, glucose did not affect ATP and ADP levels, and the ATP/ADP ratio remained stable around 7.5. In human islets, the ATP/ADP ratio progressively increased between 1 and 10 mM glucose. In duct cells, which often contaminate human islet preparations, an increase in the ATP/ADP ratio sometimes occurred between 1 and 3 mM glucose. In conclusion, the present observations establish that the regulation of glucagon secretion by glucose does not involve changes in alpha cell adenine nucleotides and further support the role of the ATP/ADP ratio in the control of insulin secretion. PMID- 9852041 TI - Abrogation of the presenilin 1/beta-catenin interaction and preservation of the heterodimeric presenilin 1 complex following caspase activation. AB - beta-Catenin has previously been shown to interact with presenilin 1 (PS1) in transfected cells. Here we report that beta-catenin co-immunoprecipitates with the endogenous C-terminal fragment of presenilin 1 (PS1-CTF) but not with the endogenous CTF of presenilin 2 (PS2-CTF) in H4 human neuroglioma cells. During staurosporine (STS)-induced cell death, beta-catenin and PS1-CTF undergo a caspase-mediated cleavage. After 12 h of STS treatment, the beta-catenin.PS1-CTF interaction is abrogated. While PS1-CTF immunoprecipitated with all caspase cleaved species of beta-catenin, beta-catenin holoprotein did not co immunoprecipitate with the "alternative" caspase-derived PS1-CTF (PS1-aCTF). Thus, the abrogation of the beta-catenin.PS1-CTF complex was due to caspase cleavage of PS1-CTF. beta-Catenin co-immunoprecipitated with PS1-NTF, but only when PS1-NTF was associated with PS1-CTF. Even though PS1-NTF.CTF complex stability was not altered by caspase cleavage, its ability to bind beta-catenin was abolished. Thus, while the PS1-NTF.CTF complex is preserved after caspase cleavage, it may no longer be fully functional. PMID- 9852042 TI - A mutational analysis of the baculovirus inhibitor of apoptosis Op-IAP. AB - A family of antiapoptotic regulators known as inhibitors of apoptosis (IAPs) was initially identified and functionally described in baculoviruses, and IAP homologues are now known in insects, birds, and mammals. Baculovirus and Drosophila IAPs inhibit apoptosis induced by Drosophila proapoptotic proteins Reaper, HID, and GRIM and physically interact with them through their baculovirus IAP repeat (BIR) region. Here we examined the functional importance of BIR and RING finger motifs of Orgyia pseudotsugata nuclear polyhedrosis virus Op-IAP and D-IAP1 in binding to and inhibiting HID. In the absence of both the BIR1 and RING motifs, the BIR2 regions of Op-IAP and D-IAP1 were able to associate with HID and block HID-induced apoptosis. Mutation of conserved amino acid residues within the BIR and RING finger motifs revealed that the conserved residues within BIR2 were essential for Op-IAP to inhibit apoptosis. However, most of the conserved residues of the BIR2 were not required for HID binding. A region at the carboxy proximal end of BIR2 was essential for the association of Op-IAP with HID. Thus binding to HID is necessary but not sufficient to block HID-induced apoptosis: the conserved residues within BIR2 must have an additional role in blocking apoptosis. These findings demonstrate that the region encompassing a single BIR of Op-IAP and D-IAP1 can be sufficient for physical interaction with and inhibition of apoptosis induced by HID. PMID- 9852043 TI - The inositol phosphatase SHIP inhibits Akt/PKB activation in B cells. AB - The serine-threonine kinase Akt/PKB is activated downstream of phosphatidylinositol 3-kinase in response to several growth factor stimuli and has been implicated in the promotion of cell survival. Although both phosphatidylinositol 3,4,5-trisphosphate (PIP3) and phosphatidylinositol 3,4 bisphosphate (PI 3,4-P2) have been implicated in the regulation of Akt activity in vitro, the relative roles of these two phospholipids in vivo are not well understood. Co-ligation of the B cell receptor (BCR) and the inhibitory FcgammaRIIB1 on B cells results in the recruitment of the 5'-inositol phosphatase SHIP to the signaling complex. Since SHIP is known to cleave PIP3 to generate PI 3,4-P2 both in vivo and in vitro, and Akt activity has been reported to be regulated by either PIP3 or PI 3,4-P2, we hypothesized that recruitment of SHIP through FcgammaRIIB1 co-cross-linking to the BCR in B cells might regulate Akt activity. The nature of this regulation, positive or negative, might also reveal the relative contribution of PIP3 and PI 3,4-P2 to Akt activation in vivo. Here we report that Akt is activated by stimulation through the BCR in a phosphatidylinositol 3-kinase-dependent manner and that this activation is inhibited by co-cross-linking of the BCR to FcgammaRIIB1. Using mutants of FcgammaRIIB1 and SHIP-deficient B cells, we demonstrate that inhibition of Akt activity is mediated by the immune cell tyrosine-based inhibitory motif within FcgammaRIIB1 as well as SHIP. The SHIP-dependent inhibition of Akt activation also suggests that PIP3 plays a greater role in Akt activation than PI 3,4-P2 in vivo. PMID- 9852044 TI - Molecular cloning, expression, and characterization of novel human SULT1C sulfotransferases that catalyze the sulfonation of N-hydroxy-2 acetylaminofluorene. AB - Upon sulfonation, carcinogenic hydroxyarylamines such as N-hydroxy-2 acetylaminofluorene (N-OH-2AAF) can be further activated to form ultimate carcinogens in vivo. Previous studies have shown that a SULT1C1 sulfotransferase is primarily responsible for the sulfonation of N-OH-2AAF in rat liver. In the present study, two novel human sulfotransferases shown to be members of the SULT1C sulfotransferase subfamily based on sequence analysis have been cloned, expressed, and characterized. Comparisons of the deduced amino acid sequence encoded by the human SULT1C sulfotransferase cDNA 1 reveal 63.7, 61.6, and 85.1% identity to the amino acid sequences of rat SULT1C1 sulfotransferase, mouse SULT1C1 sulfotransferase, and rabbit SULT1C sulfotransferase. In contrast, the deduced amino acid sequence of the human SULT1C sulfotransferase 2 cDNA displays 62.9, 63.1, 63.1, and 62.5% identity to the amino acid sequences of the human SULT1C sulfotransferase 1, rat SULT1C1 sulfotransferase, mouse SULT1C1 sulfotransferase, and rabbit SULT1C sulfotransferase. Recombinant human SULT1C sulfotransferases 1 and 2, expressed in Escherichia coli and purified to near electrophoretic homogeneity, were shown to cross-react with the antiserum against the rat liver SULT1C1 sulfotransferase and exhibited sulfonating activities with N-OH-2AAF as substrate. Tissue-specific expression of these novel human SULT1C sulfotransferases were examined by employing the Northern blotting technique. The results provide a foundation for the investigation into the functional relevance of these new SULT1C sulfotransferases in different human tissues/organs. PMID- 9852045 TI - A single amino acid in the DNA binding regions of STAT5A and STAT5B confers distinct DNA binding specificities. AB - STAT5A and STAT5B are two highly related transcription factors encoded by two distinct genes. STAT5A and STAT5B are activated by a broad range of cytokines and growth factors. Although they can be differentially activated, the functional difference between these two molecules relative to their structure is not known. Here we demonstrated that STAT5A and STAT5B homodimers have distinct DNA binding preferences. Chimeric STAT5 molecules allowed us to identify a region between amino acid 420 and 545 responsible for the DNA binding specificity. This region is located in the previously characterized DNA binding region of STAT proteins. Sequence comparison between STAT5A and STAT5B from different species showed a difference of 5 amino acids in the region 420-545 between STAT5A and STAT5B. Substitution of these amino acids demonstrated that a glycine residue at position 433 in STAT5B and a glutamic residue at a similar position in STAT5A determined the DNA binding specificity. These data indicate that STAT5A and STAT5B homodimers may have distinct function and probably regulate the expression of common as well as distinct genes. PMID- 9852046 TI - Activation of mitochondria and release of mitochondrial apoptogenic factors by betulinic acid. AB - Different classes of anticancer drugs may trigger apoptosis by acting on different subcellular targets and by activating distinct signaling pathways. Here, we report that betulinic acid (BetA) is a prototype cytotoxic agent that triggers apoptosis by a direct effect on mitochondria. In isolated mitochondria, BetA directly induces loss of transmembrane potential independent of a benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone-inhibitable caspase. This is inhibited by bongkrekic acid, an agent that stabilizes the permeability transition pore complex. Mitochondria undergoing BetA-induced permeability transition mediate cleavage of caspase-8 (FLICE/MACH/Mch5) and caspase-3 (CPP32/Yama) in a cell-free system. Soluble factors such as cytochrome c or apoptosis-inducing factor released from BetA-treated mitochondria are sufficient for cleavage of caspases and nuclear fragmentation. Addition of cytochrome c to cytosolic extracts results in cleavage of caspase-3, but not of caspase-8. However, supernatants of mitochondria, which have undergone permeability transition, and partially purified apoptosis-inducing factor activate both caspase-8 and caspase-3 in cytosolic extracts and suffice to activate recombinant caspase-8. These findings show that induction of mitochondrial permeability transition alone is sufficient to trigger the full apoptosis program and that some cytotoxic drugs such as BetA may induce apoptosis via a direct effect on mitochondria. PMID- 9852047 TI - Oligomerization state of water channels and glycerol facilitators. Involvement of loop E. AB - The major intrinsic protein (MIP) family includes water channels aquaporins (AQPs) and facilitators for small solutes such as glycerol (GlpFs). Velocity sedimentation on sucrose gradients demonstrates that heterologous AQPcic expressed in yeast or Xenopus oocytes behaves as an homotetramer when extracted by n-octyl beta-D-glucopyranoside (OG) and as a monomer when extracted by SDS. We performed an analysis of GlpF solubilized from membranes of Escherichia coli or of mRNA-injected Xenopus oocytes. The GlpF protein extracted either by SDS or by nondenaturing detergents, OG and Triton X-100, exhibits sedimentation coefficients only compatible with a monomeric form of the protein in micelles. We then substituted in loop E of AQPcic two amino acids predicted to play a role in the functional/structural properties of the MIPs. In two expression systems, yeast and oocytes, the mutant AQPcic-S205D is monomeric in OG and in SDS. The A209K mutation does not modify the tetrameric form of the heterologous protein in OG. This study shows that the serine residue at position 205 is essential for AQPcic tetramerization. Because the serine in this position is highly conserved among aquaporins and systematically replaced by an acid aspartic in GlpFs, we postulate that glycerol facilitators are monomers whereas aquaporins are organized in tetramers. Our data suggest that the role of loop E in MIP properties partly occurs through its ability to allow oligomerization of the proteins. PMID- 9852048 TI - 1alpha,25-dihydroxy-vitamin-D3-induced store-operated Ca2+ influx in skeletal muscle cells. Modulation by phospholipase c, protein kinase c, and tyrosine kinases. AB - In skeletal muscle cells the steroid hormone 1alpha, 25-dihydroxy-vitamin-D3 (1,25(OH)2D3) nongenomically promotes Ca2+ release from intracellular stores and cation influx through both L-type and store-operated Ca2+ (SOC) channels. In the present work we evaluated the regulation and kinetics of the 1, 25(OH)2D3 stimulated SOC influx in chick muscle cells. Stimulation with 10(-9) M 1,25(OH)2D3 in Ca2+-free medium resulted in a rapid (40-60 s) but transient [Ca2+]i rise, which correlated with sterol-dependent inositol 1,4,5-trisphosphate production. The SOC influx stimulated by the hormone was insensitive to both L type channel antagonists and polyphosphoinositide-specific phospholipase C (PPI PLC) inhibitors but was fully inhibitable by La3+ and Ni2+. PPI-PLC blockade prior to 1,25(OH)2D3 stimulation suppressed both the [Ca2+]i transient and the SOC influx. 1,25(OH)2D3-induced SOC entry was markedly increased after 3 min of treatment (30% above basal) and then rapidly reached a steady-state level. The sterol-stimulated SOC influx was prevented by protein kinase C and tyrosine kinase inhibitors but unaffected by blockade of the protein kinase A pathway. None of these inhibitors altered the thapsigargin-induced SOC entry, suggesting the operation of a signaling mechanism different from that for sterol-dependent SOC influx. The present results indicate that 1,25(OH)2D3-induced activation of PPI-PLC is upstream to Ca2+ influx through SOC channels and point for a role of both protein kinase C and tyrosine kinases but not protein kinase A in the regulation of the sterol-dependent SOCE pathway. PMID- 9852049 TI - Artificial chaperone-assisted refolding of citrate synthase. AB - The power of genetic engineering methods, along with increasing genomic information, makes heterologous expression of proteins an extremely important biochemical tool. Unfortunately, proteins obtained in this way often are not in their native form, and folding becomes a crucial step in protein production. We have recently developed a strategy that promotes the folding of chemically denatured proteins via the sequential addition of low molecular weight "artificial chaperones." Here we describe in detail the application of this method to porcine heart citrate synthase. Refolding yields of as high as 65% have been achieved. Mechanistic studies indicate that there are significant differences between artificial chaperone-assisted refolding of citrate synthase and artificial chaperone-assisted refolding of two other proteins that have been examined, carbonic anhydrase B (Rozema, D., and Gellman, S. H. (1996) J. Biol. Chem. 271, 3478-3487) and lysozyme (Rozema, D., and Gellman, S. H. (1996) Biochemistry 35, 15760-15771). The differences among these three test proteins reveal the range of procedural variation that must be considered in the application of the artificial chaperone method to new proteins. PMID- 9852050 TI - Generation of superoxide anion by succinate-cytochrome c reductase from bovine heart mitochondria. AB - Production of superoxide anion (O-2), measured as the chemiluminescence of the 2 methyl-6-(p-methoxyphenyl)-3, 7-dihydroimidazo[1,2-a]pyrazin-3-one hydrochloride (MCLA)-O-2 adduct, was observed during electron transfer from succinate to cytochrome c by reconstituted succinate-cytochrome c reductase-phospholipid vesicles replenished with succinate dehydrogenase. Addition of carbonyl cyanide p trifluoromethoxyphenylhydrazone or detergent to the reconstituted reductase phospholipid vesicles abolished O-2 production, suggesting that O-2 generation is caused by the membrane potential generated during electron transfer through the cytochrome bc1 complex. Production of O-2 was also observed during electron transfer from succinate to cytochrome c by antimycin-treated reductase, in which approximately 99.7% of the reductase activity was inhibited. The rate of O-2 production was closely related to the rate of antimycin-insensitive cytochrome c reduction. Factors affecting antimycin-insensitive reduction of cytochrome c also affected O-2 production and vice versa. When the oxygen concentration in the system was decreased, the rate of O-2 production and cytochrome c reduction by antimycin-treated reductase decreased. When the concentrations of MCLA and cytochrome c were increased, the rate of O-2 production and cytochrome c reduction by antimycin-treated reductase increased. The rate of antimycin insensitive cytochrome c reduction was sensitive to Qo site inhibitors such as 5 undecyl-6-hydroxy-4,7-dioxobenzothiazole. These results indicate that generation of O-2 during the oxidation of ubiquinol by the cytochrome bc1 complex results from a leakage of the second electron of ubiquinol from its Q cycle electron transfer pathway to interact with oxygen. The electron-leaking site is located at the reduced cytochrome b566 or ubisemiquinone of the Qo site because addition of MCLA to antimycin-treated cytochrome bc1 complex, in the presence of catalytic amounts of succinate-cytochrome c reductase, delayed cytochrome b reduction by succinate. In the presence of oxidized cytochrome c, purified succinate dehydrogenase also catalyzed oxidation of succinate to generate O-2. When succinate dehydrogenase was reconstituted with the bc1 particles to form succinate-cytochrome c reductase, the production of O-2 diminished. These results suggest that reduced FAD of succinate dehydrogenase is the electron donor for oxygen to produce O-2 in the absence of their immediate electron acceptor and in the presence of cytochrome c. PMID- 9852051 TI - A gene-targeted mouse model for familial hypobetalipoproteinemia. Low levels of apolipoprotein B mRNA in association with a nonsense mutation in exon 26 of the apolipoprotein B gene. AB - Familial hypobetalipoproteinemia, a syndrome characterized by abnormally low plasma levels of low density lipoprotein cholesterol, is caused by mutations in the apolipoprotein (apo) B gene that interfere with the synthesis of a full length apoB100. In many cases of familial hypobetalipoproteinemia, nonsense or frameshift mutations result in the synthesis of a truncated apoB protein. To understand why these mutations result in low plasma cholesterol levels, we used gene targeting in mouse embryonic stem cells to introduce a nonsense mutation (N1785Stop) into exon 26 of the mouse Apob gene. The sole product of this mutant Apob allele was a truncated apoB, apoB39. Mice homozygous for this "apoB39-only" (Apob39) allele had low plasma levels of apoB39 and markedly reduced plasma levels of very low density lipoprotein and low density lipoprotein cholesterol when fed a high fat diet. Analysis of liver and intestinal RNA from heterozygous apoB39-only mice revealed that the Apob39 mRNA levels were 60-70% lower than those from the wild-type allele. Interestingly, apoB39 was not cleared as rapidly from the plasma as apoB48. The apoB39-only mice provide new insights into the mechanisms of familial hypobetalipoproteinemia and the structural features of apoB that are important for lipoprotein metabolism. PMID- 9852052 TI - Physical and functional interaction between p72(syk) and erythropoietin receptor. AB - Erythropoietin (Epo) regulates the proliferation and differentiation of erythroid cells through interaction with a cell surface receptor (EpoR) that belongs to the cytokine receptor family. The Jak2 tyrosine kinase was previously shown to bind to the EpoR, to be activated upon Epo stimulation, and to play a critical role in Epo-induced proliferation. However, little is known about the role of other tyrosine kinases in Epo signaling. In this paper, we examined whether Syk was involved in EpoR activation. Coimmunoprecipitation experiments showed that the phosphorylated EpoR was associated with the Syk kinase in activated UT7 cells. The interaction of Epo with its receptor led to an increased kinase activity. The use of recombinant Syk Src homology 2 (SH2) domains expressed in tandem or individually revealed that both N- and C-SH2 domains of Syk participated in EpoR binding with a major contribution of the C-terminal SH2 domain. Far Western blotting further indicated that Syk directly binds to the EpoR and that the interaction of Syk with EpoR only occurred after Epo activation. These data suggest that phosphorylation of EpoR on tyrosine residues may mediate Syk binding to the receptor through interaction between the two SH2 domains of Syk and tyrosines of the receptor. We propose that in addition to Jak2, Syk protein kinase may be a component of EpoR signaling. PMID- 9852053 TI - Drosophila Rrp1 domain structure as defined by limited proteolysis and biophysical analyses. AB - Drosophila Rrp1 is a DNA repair nuclease whose C-terminal region shares extensive homology with Escherichia coli exonuclease III, has nuclease activity, and provides resistance to oxidative and alkylating agents in repair-deficient E. coli strains. The N-terminal 421 amino acid region of Rrp1, which binds and renatures homologous single-stranded DNA, does not share homology with any known protein. Proteolysis by endoproteinase Glu-C (protease V8) reduces the Rrp1 protein to a single, cleavage-resistant peptide. The peptide (referred to as Rrp1 C274) begins with the sequence TKTTV, corresponding to cleavage between Glu-405 and Thr-406 of Rrp1. We determined that nuclease activity is intrinsic to Rrp1 C274 although altered when compared with Rrp1; 3'-exonuclease activity is reduced 210-fold, 3'-phosphodiesterase activity is reduced 6.8-fold, and no difference in apurinic/apyrimidinic endonuclease activity is observed. Rrp1 and Rrp1-C274 are both monomers with frictional coefficients of 2.2 and 1.4, respectively. Circular dichroism results indicate that Rrp1-C274 is predominantly alpha-helical, while the N-terminal 399 amino acids is predominantly random coil. These results suggest that Rrp1 may have a bipartite structural organization; a highly organized, globular C-terminal domain; and an asymmetric, protease-sensitive random coil-enriched N-terminal region. A shape model for this bipartite structure is proposed and discussed. PMID- 9852054 TI - Sp1 and CREB mediate gastrin-dependent regulation of chromogranin A promoter activity in gastric carcinoma cells. AB - Chromogranin A (CgA) is a multifunctional acidic protein that in the stomach is expressed predominantly in enterochromaffin-like cells (ECL cells) where it is regulated by gastrin. In order to investigate the transcriptional response of the mouse CgA (mCgA) promoter to gastrin stimulation, we studied a 4.8-kilobase mCgA promoter-luciferase reporter gene construct in transiently transfected AGS-B cells. 5'-Deletion analysis and scanning mutagenesis of mCgA 5'-flanking DNA showed that a Sp1/Egr-1 site spanning -88 to -77 base pairs (bp) and a cyclic AMP responsive element (CRE) at -71 to -64 bp are essential for gastrin-dependent mCgA transactivation. Gastrin stimulation increased cellular Sp1 protein levels and Sp1-binding to the mCgA -88 to -77 bp element, as well as binding of CREB to its consensus motif at -71 to -64 bp. Gastrin also stimulated CREB Ser-133 phosphorylation, and abundance of cellular CREB protein levels. Overexpression of either Sp1 or phosphorylated CREB transactivated the mCgA promoter dose dependently, while coexpression of both transcription factors resulted in an additive mCgA promoter response. mCgA -92 to -64 bp, comprising the Sp1/Egr-1 site and the CRE motif, conferred gastrin responsiveness to a heterologous thymidine kinase promoter system, and therefore functions as a "true" enhancer element. This report demonstrates that Sp1 and CREB mediate CCK-B/gastrin receptor-dependent gene regulation, and that the effect of gastrin on the CgA gene is brought about by cooperative action of both transcription factors. PMID- 9852055 TI - Probing the function of the invariant glutamyl residue 312 in spinach ferredoxin NADP+ reductase. AB - Ferredoxin-NADP+ reductase, the prototype of a large family of structurally related flavoenzymes, pairs single electrons carried by ferredoxin I and transfers them as a hydride to NADP+. Four mutants of the enzyme, in which Glu 312 was replaced with Asp, Gln, Leu, and Ala to probe the role of the residue charge, size, and polarity in the enzyme activity, have been heterologously expressed, purified, and characterized through steady-state, rapid kinetic studies, ligand-binding experiments, and three-dimensional structure determination by x-ray crystallography. The E312L mutant was the only one that was almost inactive (approximately 1%), whereas unexpectedly the E312A reductase was 10-100% active with the various acceptors tested. Rapid kinetic absorption spectroscopy studies demonstrated that flavin reduction by NADPH was impaired in the mutants. Furthermore, NADP(H) binding was partially perturbed. These functional and structural studies lead us to conclude that Glu-312 does not fulfil the role of proton donor during catalysis, but it is required for proper binding of the nicotinamide ring of NADP(H). In addition, its charge modulates the two one-electron redox potentials of the flavin to stabilize the semiquinone form. PMID- 9852056 TI - Arg278, but not Lys229 or Lys236, plays an important role in the binding of retinoic acid by retinoic acid receptor gamma. AB - The diverse biological actions of retinoic acid (RA) are mediated by retinoic acid receptors (RARalpha, beta and gamma) and retinoid X receptors (RXR alpha, beta, and gamma). Although the ligand-binding domains of RARs share the same novel folding pattern, many RAR subtype-specific retinoids have been synthesized indicating that the ligand-binding pocket of each RAR subtype has unique features. Previously we have demonstrated the importance for RA binding and RA dependent transactivation of Arg276 of RARalpha alone and in RARbeta Arg269 in conjunction with Lys220. In this study, we have examined the role of the homologous amino acid residues (Lys229 and Arg278) in RARgamma for these activities. Like RARalpha but dissimilar to RARbeta, Arg278 in RARgamma alone was found to play an important role in RA binding and RA-dependent transactivation. Since Lys236 in RARgamma was suggested from the crystal structure of holo RARgamma to interact with RA, we also examined its role and that of its homologs in RARalpha and RARbeta. Despite the suggestion from the crystal structure, neither Lys236 nor its homologs in RARalpha and RARbeta play a role in the binding of RA or RA-dependent transactivation. It is likely that Lys236 in RARgamma and its homologs in RARalpha and RARbeta are solvent exposed rather than pointing into the RA-binding pocket. PMID- 9852057 TI - Contributions to maxima in protein kinase C activation. AB - In many lipid systems, the activity of protein kinase C (PKC) exhibits a peak followed by a decline as the mol % of one component is increased. In these systems, an increase in one lipid component is always at the expense of another or accompanied by a change in total lipid concentration. Here we report that in saturated phosphatidylserine (PS)/phosphatidylcholine (PC)/diacylglycerol (DAG) mixtures, increasing PS or DAG at the expense of PC revealed an optimal mol % PS, dependent on mol % DAG, with higher mol % PS diminishing activity. The decrease at high mol % PS is probably not attributable simply to more gel-phase lipid due to the higher melting temperature of saturated PS versus PC because a similar peak in activity occurred in unsaturated lipid systems. Increasing the total lipid concentration at suboptimal mol % PS provided the same activity as higher mol % PS at lower total lipid concentration. However, at optimal mol % PS, activity increased and then decreased as a function of total lipid concentration. PKC autophosphorylation also exhibited an optimum as a function of mol % PS, and increasing the PKC concentration increased the mol % PS at which activity decreased, both for autophosphorylation and for heterologous phosphorylation. Formation of two-dimensional crystals of PKC on lipid monolayers also exhibited a peak as a function of mol % PS, and the unit cell size of the crystals formed shifts from 50 x 50 A at low mol % PS to 75 x 75 A at higher PS. Collectively, these data suggest the existence of optimal lipid compositions for PKC activation, with increased quantity of these domains serving to dilute out enzyme substrate aggregates and/or enzyme-enzyme aggregates on the lipid surface. PMID- 9852058 TI - Regulation of the Salmonella typhimurium flavohemoglobin gene. A new pathway for bacterial gene expression in response to nitric oxide. AB - Flavohemoglobins, a family of two-domain proteins with homology to vertebrate hemoglobins, are found in a variety of prokaryotic and eukaryotic microorganisms. Recent studies suggest a role for these proteins in nitrogen oxide metabolism. We now show that nitric oxide donors positively regulate a chromosomal flavohemoglobin (hmp)/lacZ operon fusion in Salmonella typhimurium. hmp gene expression in the presence of NO. is independent of the SoxS, OxyR, and FNR transcription factors and instead relies on inactivation of the iron-dependent Fur repressor. Other Fur-repressed promoters in S. typhimurium are also activated by an NO. donor. In contrast to the wild-type strain, an hmp- mutant requires markedly lower concentrations of NO to induce the hmp/lacZ fusion, whereas its response to iron chelation is equivalent to wild type. These data unveil a new pathway for NO-dependent gene expression in S. typhimurium. PMID- 9852059 TI - Evidence for the location of bicyclomycin binding to the Escherichia coli transcription termination factor Rho. AB - The commercial antibiotic bicyclomycin (Bcm) has been shown to target the essential transcription termination factor Rho in Escherichia coli. Little is known about the Bcm binding domain in Rho. A recent structure-activity relationship study led us to evaluate the reductive amination probe, 5a-(3 formylanilino)dihydrobicyclomycin (FD-Bcm). Biochemical studies showed that FD Bcm possessed inhibitory activities comparable to Bcm in Rho-dependent ATPase and transcription termination assays. Incubation of Rho with FD-Bcm, ATP, and poly(C) followed by NaBH4 reduction and dialysis led to an appreciable loss of ATPase activity. Inclusion of Bcm with FD-Bcm in the reductive amination reaction protected Rho, indicating that Bcm and FD-Bcm competed for the same binding site in Rho. Incubation of Rho with FD-Bcm and poly(C) followed by NaBH4 reduction provided a sample with residual ATPase activity (12%). Mass spectrometric analysis indicated the presence of two proteins in an approximate 1.2:1 ratio, whose masses corresponded to wild-type Rho (47,010 Da) and lysine-modified Rho (47,417 Da), respectively. Trypsin digestion of the Rho sample followed by high performance liquid chromatography separation and tandem mass spectrometry analysis identified the site of modification as Lys181 within the combined tryptic fragment, Gly-Leu-Ile-Val-Ala-Pro-Pro-Lys-Ala-Gly-Lys (residues 174-184). Similar analysis of a lesser modified sample (following incubation with inclusion of ATP) showed that addition had again occurred at Lys181. These findings provide the first structural information concerning the site of Bcm binding in Rho. PMID- 9852060 TI - Abortive initiation of transcription at a hybrid promoter. An analysis of the sliding clamp activator of bacteriophage T4 late transcription, and a comparison of the sigma70 and T4 gp55 promoter recognition proteins. AB - Bacteriophage T4 late promoters are transcribed by an RNA polymerase holoenzyme comprising the Escherichia coli core, E, the phage gene 55-encoded promoter recognition subunit, gp55, and the gene 33-encoded co-activator, gp33. Transcriptional initiation is activated by the T4 gene 45-encoded sliding clamp, which is loaded on to DNA at enhancer-like sites by its clamp-loader. Correct initiation of transcription at late promoters in basal mode requires only RNA polymerase core and gp55 (E.gp55). Dinucleotide-primed abortive initiation of basal and activated T4 late transcription has been compared. Only the trinucleotide non-productive transcript is made at a high rate; all other short transcripts are made at rates of less than one molecule per productive transcript. Gp45 increases abortive trinucleotide synthesis along with productive transcription, although the proportion of productive transcripts is also elevated. Nevertheless, this increase accounts for only a small part of the activation of T4 late transcription that is generated by its activator and co activator. The pattern of production of short transcripts differs subtly between basal and enhanced transcription, indicating that linking the RNA polymerase with its sliding clamp activator only generates minor changes in the transition from abortive to productive RNA chain elongation. The T4 late promoter is converted to a strong sigma70 promoter by inserting an appropriate -35 promoter element. A direct comparison at such a hybrid promoter shows sigma70 and gp55 generating qualitatively and quantitative different patterns of abortive initiation at the same start site. PMID- 9852061 TI - Binding of 9-anthroylcholine monitors the interactions of adenosine cyclic 3',5' phosphate-dependent protein kinase with MgATP, substrates, and regulatory subunits. AB - The isolated catalytic subunit of cAMP-dependent protein kinase and smooth muscle myosin light chain kinase undergo interactions with the fluorescent dye 9 anthroylcholine (9AC) that are responsive to the two enzymes' associations with substrates and effectors. Additionally, the binding of 9AC is highly sensitive to subtle structural or functional differences among closely related protein kinases. Skeletal muscle myosin light chain kinase and the catalytically active chymotryptic fragment of the gamma-subunit of phosphorylase kinase do not associate with 9AC. The 1:1 fluorescent complex of the isolated catalytic subunit of cAMP-dependent protein kinase with 9AC exhibits a dissociation constant of 21 microM. The association of the catalytic subunit with either of the regulatory subunits, RI and RII, results in decreases in the observed 9AC fluorescence that are reversed upon the addition of cAMP. The effects of MgATP and of polypeptide substrates (Kemptide, troponin I, protamine) on the 9AC-catalytic subunit complex are consistent with a general noncompetitive model in which the interactions of 9AC and the other ligands with the enzyme are mutually antagonistic but not purely competitive. PMID- 9852062 TI - Identification of Glu-540 as the catalytic nucleophile of human beta glucuronidase using electrospray mass spectrometry. AB - Human beta-glucuronidase is a member of the Family 2 glycosylhydrolases that cleaves beta-D-glucuronic acid residues from the nonreducing termini of glycosaminoglycans. The enzyme is shown to catalyze glycoside bond hydrolysis with net retention of anomeric configuration, presumably via a mechanism involving a covalent glucuronyl-enzyme intermediate. Incubation of human beta glucuronidase with 2-deoxy-2-fluoro-beta-D-glucuronyl fluoride resulted in time dependent inactivation of the enzyme through the accumulation of a covalent 2 deoxy-2-fluoro-alpha-D-glucuronyl-enzyme, as observed by electrospray mass spectrometry. Regeneration of the free enzyme by hydrolysis or transglycosylation and removal of excess inactivator demonstrated that the covalent intermediate was kinetically competent. Peptic digestion of the 2-deoxy-2-fluoro-alpha-D glucuronyl-enzyme intermediate and subsequent analysis by liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry indicated the presence of a 2-deoxy-2-fluoro-alpha-D-glucuronyl peptide. Sequence determination of the labeled peptide by tandem mass spectrometry in the daughter ion scan mode permitted the identification of Glu-540 as the catalytic nucleophile within the sequence SEYGAET. PMID- 9852063 TI - Acute suppression of inwardly rectifying Kir2.1 channels by direct tyrosine kinase phosphorylation. AB - Signaling via cytosolic and receptor tyrosine kinases is associated with cell growth and differentiation but also targets onto transmitter receptors and ion channels. Here, regulation by tyrosine kinase (TK) activity was investigated for inwardly rectifying K+ (Kir2.1) channels that control membrane excitability in many central neurons. In mammalian tsA-201 cells, the membrane-permeable protein tyrosine phosphatase inhibitor, perorthovanadate (100 microM), suppressed currents through recombinant Kir2.1 channels by 60 +/- 20%. Coapplication of the TK inhibitor genistein (100 microM) completely abolished this effect. Native Kir2.1 channels in rat basophilic leukocytes were affected by manipulation of the TK and protein tyrosine phosphatase activity in a qualitatively similar manner. Site mutation of a tyrosine consensus residue for TK phosphorylation in the C terminal domain of Kir2.1 generated channel properties indistinguishable from wild-type Kir2.1 channels. However, Kir2.1Y242F channels were no longer suppressed following exposure to perorthovanadate, indicating that the channel is a direct substrate for TKs. After coexpression of nerve growth factor receptor with Kir2.1 channels in tsA-201 cells and Xenopus oocytes, the activity of Kir2.1 was rapidly suppressed by applied nerve growth factor (0.5 microgram/ml) by 31 +/ 10 and 21 +/- 15%, respectively. Acute inhibition was also evoked by epidermal growth factor and insulin via endogenous insulin receptors, indicating that Kir2.1 channels may serve as a general target for neurotrophic growth factors in the brain. PMID- 9852064 TI - MinK-KvLQT1 fusion proteins, evidence for multiple stoichiometries of the assembled IsK channel. AB - IsK, a slowly activating delayed rectifier K+ current through channels formed by the assembly of two channel proteins KvLQT1 and MinK, modulates the repolarization of cardiac action potentials. Mutations that map to the KvLQT1 and minK genes account for more than 50% of an inherited cardiac disorder, the Long QT syndrome (Splawski, I., Tristani-Firouzi, M., Lehmann, M. H., Sanguinetti, M. C., and Keating, M. T. (1997) Nat. Genet. 17, 338-340). Despite the importance of these channels to human cardiac function, the molecular basis of their uniquely slow gating properties as well as the stoichiometry and interaction sites of these two subunits are still unclear. We have constructed several fusion channel proteins to begin investigating the stoichiometry of these two subunits and the role of voltage-dependent subunit assembly in channel gating. Functional properties of these constructs were measured using whole cell patch clamp recordings of transiently transfected Chinese hamster ovary cells. The constructs we tested are as follows: MK24 (C terminus of MinK linked to N terminus of KvLQT1); KK40 (a tandem homodimer of KvLQT1); and MKK44 (C terminus of MinK linked to N terminus of KK40). In control experiments (no DNA, control DNA, or only MinK), no time-dependent K+ current was observed. Expression of KvLQT1 or KK40 produced currents that activate and inactivate in a voltage-dependent manner as reported by others for KvLQT1. In contrast, expression of MK24 and MKK44 elicited current with activation kinetics and voltage dependence very similar to native IsK and identical to currents expressed by cells co-transfected with independent MinK and KvLQT1 cDNA. Expression of MK24 plus additional MinK significantly slows current kinetics. Our data raise the possibility 1) of multiple MinK/KvLQT1 stoichiometries and 2) indicate that uniquely slow kinetics of IsK channels is due to voltage-dependent conformational changes of the channel protein and not to assembly of channel subunits. PMID- 9852065 TI - Substrate mutations that bypass a specific Cpn10 chaperonin requirement for protein folding. AB - The bacteriophage T4 GroES homologue, gp31, in conjunction with the Escherichia coli chaperonin GroEL, is both necessary and sufficient to fold the T4 major capsid protein, gp23, to a state competent for capsid assembly as shown by in vivo expression studies. GroES is unable to function in this role as a productive co-chaperonin. The sequencing and characterization of mutations within gp23 that confer GroEL and gp31 chaperonin-independent folding of the mutant protein suggest that the chaperonin requirements are due to specific sequence determinants or structures in critical regions of gp23 that behave in an additive fashion to confer a chaperonin bypass phenotype. Conservative amino acid substitutions in these critical regions enable gp23 to fold in a GroEL-gp31 chaperonin-independent mode, albeit less efficiently than wild type, both in vivo and in vitro. Although the presence of functional GroEL-gp31 enhances folding of the mutated gp23 in vivo, GroEL-GroES has no such effect. Site-directed mutagenesis experiments suggest that a translational pausing mechanism is not responsible for the bypass mutant phenotype. Polyhead reassembly experiments are also consistent with direct, post-translational effects of the bypass mutations on polypeptide folding. Given our finding that gp31 is not required for the binding of the major capsid protein to GroEL and that active GroES is incapable of folding the gp23 polypeptide chain to native conformation, our results suggest co-chaperonin specificity in the folding of certain substrates. PMID- 9852066 TI - Phosphorylation and O-glycosylation sites of human chromogranin A (CGA79-439) from urine of patients with carcinoid tumors. AB - Because of their water-soluble properties, chromogranins (CGs) and chromogranin derived fragments are released together with catecholamines from adrenal chromaffin cells during stress situations and can be detected in the blood by radiochemical and enzyme assays. It is well known that chromogranins can serve as immunocytochemical markers for neuroendocrine tissues and as a diagnostic tool for neuroendocrine tumors. In 1993, large CGA-derived fragments have been shown to be excreted into the urine in patients with carcinoid tumors and the present study deals with the characterization of the post-translational modifications (phosphorylation and O-glycosylation) located along the largest natural CGA derived fragment CGA79-439. Using mild proteolysis of peptidic material, high performance liquid chromatography, sequencing, and mass spectrometry analysis, six post-translational modifications were detected along the C-terminal CGA derived fragment CGA79-439. Three O-linked glycosylation sites were located in the core of the protein on Thr163, Thr165, and Thr233, consisting in di-, tri-, and tetrasaccharides. Three phosphorylation sites were located in the middle and C-terminal domain, on serine residues Ser200, Ser252, and Ser315. These modified sites were compared with sequences of others species and discussed in relation with the post-translational modifications that we have reported previously for bovine CGA. PMID- 9852067 TI - Allosteric regulation of Trypanosoma brucei ribonucleotide reductase studied in vitro and in vivo. AB - Trypanosoma brucei is the causative agent for African sleeping sickness. We have made in vitro and in vivo studies on the allosteric regulation of the trypanosome ribonucleotide reductase, a key enzyme in the production of dNTPs needed for DNA synthesis. Results with the isolated recombinant trypanosome ribonucleotide reductase showed that dATP specifically directs pyrimidine ribonucleotide reduction instead of being a general negative effector as in other related ribonucleotide reductases, whereas dTTP and dGTP directed GDP and ADP reduction, respectively. Pool measurements of NDPs, NTPs, and dNTPs in the cultivated bloodstream form of trypanosomes exposed to deoxyribonucleosides or inhibited by hydroxyurea confirmed our in vitro allosteric regulation model of ribonucleotide reductase. Interestingly, the trypanosomes had extremely low CDP and CTP pools, whereas the dCTP pool was comparable with that of other dNTPs. The trypanosome ribonucleotide reductase seems adapted to this situation by having a high affinity for the CDP/UDP-specific effector dATP and a high catalytic efficiency, Kcat/Km, for CDP reduction. Thymidine and deoxyadenosine were readily taken up and phosphorylated to dTTP and dATP, respectively, the latter in a nonsaturating manner. This uncontrolled uptake of deoxyadenosine strongly inhibited trypanosome proliferation, a valuable observation in the search for new trypanocidal nucleoside analogues. PMID- 9852068 TI - Hepatocyte nuclear factor-4 controls transcription from a TATA-less human sex hormone-binding globulin gene promoter. AB - Hepatocytes are the major source of sex hormone-binding globulin (SHBG), a glycoprotein that transports sex steroids in the blood and regulates their access to target tissues. The human SHBG proximal promoter was analyzed by DNase I footprinting, and the functional significance of 6 footprinted regions (FP1-FP6) within the proximal promoter was studied in human HepG2 hepatoblastoma cells. Two footprinted regions (FP1 and FP3) contain binding sites for the chicken ovalbumin upstream promoter-transcription factor (COUP-TF) and hepatocyte nuclear factor-4 (HNF-4). In experiments where SHBG promoter-luciferase reporter gene constructs were co-transfected into HepG2 cells with COUP-TF and/or HNF-4 expression vectors, HNF-4 markedly increased transcription, whereas COUP-TF suppressed this probably by displacing HNF-4 from their common FP1-binding site. This COUP-TF/HNF 4-binding site within FP1 includes a TTTAA sequence, located at nucleotides -30/ 26 upstream of the transcription start site, which fails to interact with human TFIID, TATA-binding protein in vitro. When this sequence was replaced with an idealized HNF-4-binding site, the transcriptional activity of the promoter increased in HepG2 cells. Taken together, these data imply that an interplay between COUP-TF and HNF-4 at a site within FP1 regulates human SHBG expression and that HNF-4 controls transcription from this TATA-less promoter by somehow substituting for TATA-binding protein in the recruitment of a transcription preinitiation complex. PMID- 9852069 TI - Gene structure, expression pattern, and biological activity of mouse killer cell activating receptor-associated protein (KARAP)/DAP-12. AB - Natural killer cell and T cell subsets express at their cell surface a repertoire of receptors for MHC class I molecules, the natural killer cell receptors (NKRs). NKRs are characterized by the existence of inhibitory and activating isoforms, which are encoded by highly homologous but separate genes present in the same locus. Inhibitory isoforms express an intracytoplasmic immunoreceptor tyrosine based inhibition motif, whereas activating isoforms lack any immunoreceptor tyrosine-based inhibition motif but harbor a charged amino acid residue in their transmembrane domain. We previously characterized KARAP (killer cell activating receptor-associated protein), a novel disulfide-linked tyrosine-phosphorylated dimer that selectively associates with the activating NKR isoforms. We report here the identification of the mouse KARAP gene, its localization on chromosome 7 and its genomic organization in five exons. Point mutation and transfection studies revealed that KARAP is a novel signaling transmembrane subunit whose transduction function depends on the integrity of an intracytoplasmic immunoreceptor tyrosine-based activation motif. In contrast to previous members of the immunoreceptor tyrosine-based activation motif polypeptide family, KARAP is ubiquitously expressed on hematopoietic and nonhematopoietic cells, suggesting its association with a broad range of activating receptors in a variety of tissues. PMID- 9852070 TI - The involvement of multiple tumor necrosis factor receptor (TNFR)-associated factors in the signaling mechanisms of receptor activator of NF-kappaB, a member of the TNFR superfamily. AB - Receptor activator of NF-kappaB (RANK) is a recently identified member of the tumor necrosis factor receptor superfamily and is expressed on activated T cells and dendritic cells. Its cognate ligand (RANKL) plays significant roles in the activation of dendritic cell function and osteoclast differentiation. We demonstrate here the interaction of RANK with tumor necrosis factor receptor associated factors (TRAFs) 1, 2, 3, 5, and 6 both in vitro and in cells. Mapping of the structural requirements for TRAF/RANK interaction revealed multiple TRAF binding sites clustered in two distinct domains in the RANK cytoplasmic tail. These TRAF binding domains were shown to be functionally important for the RANK dependent induction of NF-kappaB and c-Jun NH2-terminal kinase activities. Site directed mutagenesis demonstrated that these TRAF binding sites exhibited selective binding for different TRAF proteins. In particular, TRAF6 interacted with membrane-proximal determinants distinct from those binding TRAFs 1, 2, 3, and 5. When this membrane-proximal TRAF6 interaction domain was deleted, RANK mediated NF-kappaB signaling was completely inhibited while c-Jun NH2-terminal kinase activation was partially inhibited. An NH2-terminal truncation mutant of TRAF6 inhibited RANKL-mediated NF-kappaB activation, but failed to affect constitutive signaling induced by receptor overexpression, revealing a selective role for TRAF6 in ligand-induced activation events. PMID- 9852071 TI - Activation/attenuation model for RNase H. A one-metal mechanism with second-metal inhibition. AB - Ribonucleases H (RNases H) comprise a family of metal-dependent enzymes that catalyze the hydrolysis of the 3'-O---P bond of RNA in RNA.DNA hybrids. The mechanism by which RNases H use active-site metal(s) for catalysis is unclear. Based upon the seemingly contradictory structural observations of one divalent metal bound to Escherichia coli RNase HI and two divalent metals bound to the HIV RNase H domain, two models explaining RNase H metal dependence have been proposed: a one-metal mechanism and a two-metal mechanism. In this paper, we show that the Mn2+-dependent activity of E. coli RNase HI is not consistent with either of these mechanisms. RNase H activity in the presence of Mn2+ is complex, with activation and inhibition of the enzyme at low and high Mn2+ concentrations, respectively. Mutations at Asp-134 result in a partial loss of this inhibition, with little effect on activation. Neutralization of His-124 by mutation to Ala results in an enzyme with a significantly decreased specific activity and an absolute loss of Mn2+ inhibition. Inhibition by high Mn2+ concentrations is shown to be due to a reduction in kcat; this attenuation has a critical dependence on the presence of His-124. Based upon these results, we propose an "activation/attenuation" model explaining the metal dependence of RNase H activity where one metal is required for enzyme activation and binding of a second metal is inhibitory. PMID- 9852072 TI - A new remote subsite in ribonuclease A. AB - The interaction between bovine pancreatic ribonuclease A (RNase A) and its RNA substrate extends beyond the scissile bond. Enzymic subsites interact with the bases and the phosphoryl groups of a bound substrate. We evaluated the four cationic residues closest to known subsites for their abilities to interact with a bound nucleic acid. Lys-37, Arg-39, Arg-85, and Lys-104 were replaced individually by an alanine residue, and the resulting enzymes were assayed as catalysts of poly(cytidylic acid) (poly(C)) cleavage. The values of Km and kcat/Km for poly(C) cleavage were affected only by replacing Arg-85. Moreover, the contribution of Arg-85 to the binding of the ground state and the transition state was uniform---Km increased by 15-fold and kcat/Km decreased by 10-fold. The contribution of Arg-85 to binding was also apparent in the values of Kd for complexes with oligonucleotides of different length. This contribution was dependent on salt concentration, as expected from a coulombic interaction between a cationic side chain and an anionic phosphoryl group. Together, these data indicate that Arg-85 interacts with a particular phosphoryl group of a bound nucleic acid. We propose that Arg-85 comprises a new distal subsite in RNase A-- the P(-1) subsite. PMID- 9852073 TI - Werner syndrome protein. I. DNA helicase and dna exonuclease reside on the same polypeptide. AB - Werner Syndrome (WS) is a human progeroid disorder characterized by genomic instability. The gene defective in WS encodes a 3' --> 5' DNA helicase (Gray, M. D., Shen, J.-C., Kamath-Loeb, A. S., Blank, A. , Sopher, B. L., Martin, G. M., Oshima, J., and Loeb, L. A.(1997) Nat. Genet. 17, 100-103). Sequence alignment analysis identified an N-terminal motif in WRN that is homologous to several exonucleases. Using combined molecular genetic, biochemical, and immunochemical approaches, we demonstrate that WRN also exhibits an integral DNA exonuclease activity. First, whereas wild-type recombinant WRN possesses both helicase and exonuclease activities, mutant WRN lacking the nuclease domain does not display exonucleolytic activity. In contrast, WRN proteins with defective helicase activity are active in exonucleolytic digestion of DNA. Second, the exonuclease co-purifies with the 160-kDa WRN protein and its associated DNA helicase and ATPase activities through successive steps of ion exchange and affinity chromatography, suggesting that all three activities are physically associated. Lastly, anti-WRN antiserum specifically co-precipitates the WRN helicase and exonuclease activities indicating that both activities reside on the same antigenic WRN polypeptide. The association of an exonuclease with WRN distinguishes it from other RecQ homologs and raises the possibility that the distinct phenotypic characteristics of WS may be due in part to a defective exonuclease. PMID- 9852074 TI - Werner syndrome protein. II. Characterization of the integral 3' --> 5' DNA exonuclease. AB - In addition to its DNA helicase activity, Werner syndrome protein (WRN) also possesses an exonuclease activity (Shen, J.-C., Gray, M. D., Kamath-Loeb, A. S., Fry, M., Oshima, J., and Loeb, L. A. (1998) J. Biol. Chem. 273, 34139-34144). Here we describe the properties of nearly homogeneous WRN exonuclease. WRN exonuclease hydrolyzes a recessed strand in a partial DNA duplex but does not significantly digest single-stranded DNA, blunt-ended duplex, or a protruding strand of a partial duplex. Although DNA is hydrolyzed in the absence of nucleoside triphosphates, nuclease activity is markedly stimulated by ATP, dATP, or CTP. WRN exonuclease digests DNA with a 3' --> 5' directionality to generate 5'-dNMP products, and DNA strands terminating with either a 3'-OH or 3'-PO4 group are hydrolyzed to similar extents. A recessed DNA strand with a single 3' terminal mismatch is hydrolyzed more efficiently by WRN than one with a complementary nucleotide, but the enzyme fails to hydrolyze a DNA strand terminating with two mismatched bases. WRN exonuclease is distinguished from known mammalian DNA nucleases by its covalent association with a DNA helicase, preference for a recessed DNA strand, stimulation by ATP, ability to equally digest DNA with 3'-OH or 3'-PO4 termini, and its preferential digestion of DNA with a single 3'-terminal mismatch. PMID- 9852075 TI - Yeast and viral RNA 5' triphosphatases comprise a new nucleoside triphosphatase family. AB - Saccharomyces cerevisiae Cet1p catalyzes the first step of mRNA capping, the hydrolysis of the gamma phosphate of triphosphate-terminated RNA to form a 5' diphosphate end. The RNA triphosphatase activity of Cet1p is magnesium-dependent and has a turnover number of 1 s-1. Here we show that purified recombinant Cet1p possesses a robust ATPase activity (Km = 2.8 microM; Vmax = 25 s-1) in the presence of manganese. Cobalt is also an effective cofactor, but magnesium, calcium, copper, and zinc are not. Cet1p displays broad specificity in converting ribonucleoside triphosphates and deoxynucleoside triphosphates to their respective diphosphates. The manganese- and cobalt-dependent nucleoside triphosphatase of Cet1p resembles the nucleoside triphosphatase activities of the baculovirus LEF-4 and vaccinia virus D1 capping enzymes. Cet1p, LEF-4, and D1 share three collinear sequence motifs. Mutational analysis establishes that conserved glutamate and arginine side chains within these motifs are essential for the RNA triphosphatase and ATPase activities of Cet1p in vitro and for Cet1p function in vivo. These findings are in accord with the effects of single alanine mutations at analogous positions of vaccinia capping enzyme. We suggest that the metal-dependent RNA triphosphatases encoded by yeast and DNA viruses comprise a novel family of phosphohydrolase enzymes with a common active site. PMID- 9852076 TI - Reversible phosphorylation of Bcl2 following interleukin 3 or bryostatin 1 is mediated by direct interaction with protein phosphatase 2A. AB - Interleukin 3 (IL-3) stimulates the net growth of murine factor-dependent NSF/N1.H7 and FDC-P1/ER myeloid cells by stimulating proliferation and suppressing apoptosis. Recently, we discovered that Bcl2 is phosphorylated at an evolutionarily conserved serine residue (Ser70) after treatment with the survival agonists IL-3 or bryostatin 1, a potent activator of protein kinase (Ito, T., Deng, X., Carr, B., and May, W. S. (1997) J. Biol. Chem. 272, 11671-11673). In addition, an intact Ser70 was found to be required for Bcl2's ability to suppress apoptosis after IL-3 withdrawal or toxic chemotherapy. We now show that phosphorylation of Bcl2 occurs rapidly after the addition of agonist to IL-3 deprived cells and can be reversed by the action of an okadaic acid (OA) sensitive phosphatase. A role for protein phosphatase (PP) 2A as the Bcl2 regulatory phosphatase is supported by several observations: 1) dephosphorylation of Bcl2 is blocked by OA, a potent PP1 and PP2A inhibitor; 2) intracellular PP2A, but not PP1, co-localizes with Bcl2; 3) the purified PP2Ac catalytic subunit directly dephosphorylates Bcl2 in vitro in an OA-sensitive manner; 4) the purified PP2Ac catalytic subunit preferentially dephosphorylates Bcl2 in vitro compared with PP1 and PP2B; 5) reciprocal immunoprecipitation studies indicate a direct interaction between PP2A and hemagglutinin (HA)-Bcl2; and 6) treatment of factor-deprived cells with bryostatin 1 dramatically increases the association between PP2A and Bcl2. Increased association between Bcl2 and PP2A occurs 15 min after agonist stimulation when Bcl2 phosphorylation has peaked and immediately before dephosphorylation. An agonist-induced increased association of PP2A and Bcl2 fails to occur in cells expressing the inactive, phosphorylation-negative S70A Bcl2 mutant, which indicates that an intact Ser70 site is necessary and sufficient for the interaction to occur. Functional phosphorylation of Bcl2 at Ser70 is proposed to be a dynamic process regulated by the sequential action of an agonist-activated Bcl2 kinase and PP2A. PMID- 9852077 TI - Effects of Asp-369 and Arg-372 mutations on heme environment and function in human endothelial nitric-oxide synthase. AB - Eight polar amino acid residues in the putative substrate-binding region from Thr 360 to Val-379 in human endothelial nitric-oxide synthase (eNOS) (Thr-360, Arg 365, Cys-368, Asp-369, Arg-372, Tyr-373, Glu-377, and Asp-378) were individually mutated. Only two of these residues, Asp-369 and Arg-372, were found to be essential for enzyme activity. A further series of mutants was generated by replacing these two residues with various amino acids and the mutant proteins were expressed in a baculovirus system. Mutant eNOS had a very low L-citrulline formation activity with the exception of D369E and R372K, which retained 27% and 44% of the wild-type enzyme activity, respectively. Unlike the wild-type enzyme, all mutants except D369E, R372K, and R372M had a low spin heme (Soret peak at 416 nm). All the Asp-369 mutants had higher Kd values for L-arginine (1-10 mM) than wild-type eNOS (0.4 microM) and an unstable heme-CO complex, and except for D369E, had a very low (6R)-5,6,7, 8-tetrahydro-L-biopterin (BH4) content. In contrast, each of Arg-372 mutants retained a considerable amount of BH4, had a moderate reduction in L-arginine affinity, and had a more stable heme-CO complex. 1-Phenylimidazole did not bind to wild-type eNOS heme, but bound to all Asp-369 and Arg-372 mutants (Kd ranged from 10 to 65 microM) except R372K. Heme spin state changes caused by binding of 3, 5-lutidine appeared to depend on both charge and size of the side chains of residues 369 and 372. Furthermore, all Asp 369 and Arg-372 mutants were defective in dimer formation. These results suggest that residues Asp-369 and Arg-372 in eNOS play a critical role in oxygenase domain active-site structure and activity. PMID- 9852078 TI - Three novel proteins of the syntaxin/SNAP-25 family. AB - Intracellular membrane traffic is thought to be regulated in part by soluble N ethylmaleimide-sensitive factor-attachment protein receptors (SNAREs) through the formation of complexes between these proteins present on vesicle and target membranes. All known SNARE-mediated fusion events involve members of the syntaxin and vesicle-associated membrane protein families. The diversity of mammalian membrane compartments predicts the existence of a large number of different syntaxin and vesicle-associated membrane protein genes. To further investigate the spectrum of SNAREs and their roles in membrane trafficking we characterized three novel members of the syntaxin and SNAP-25 (synaptosome-associated protein of 25 kDa) subfamilies. The proteins are broadly expressed, suggesting a general role in vesicle trafficking, and localize to distinct membrane compartments. Syntaxin 8 co-localizes with markers of the endoplasmic reticulum. Syntaxin 17, a divergent member of the syntaxin family, partially overlaps with endoplasmic reticulum markers, and SNAP-29 is broadly localized on multiple membranes. SNAP 29 does not contain a predicted membrane anchor characteristic of other SNAREs. In vitro studies established that SNAP-29 is capable of binding to a broad range of syntaxins. PMID- 9852079 TI - Affinity purification and partial characterization of a yeast multiprotein complex for nucleotide excision repair using histidine-tagged Rad14 protein. AB - The nucleotide excision repair (NER) pathway of eukaryotes involves approximately 30 polypeptides. Reconstitution of this pathway with purified components is consistent with the sequential assembly of NER proteins at the DNA lesion. However, recent studies have suggested that NER proteins may be pre-assembled in a high molecular weight complex in the absence of DNA damage. To examine this model further, we have constructed a histidine-tagged version of the yeast DNA damage recognition protein Rad14. Affinity purification of this protein from yeast nuclear extracts resulted in the co-purification of Rad1, Rad7, Rad10, Rad16, Rad23, RPA, RPB1, and TFIIH proteins, whereas none of these proteins bound to the affinity resin in the absence of recombinant Rad14. Furthermore, many of the co-purifying proteins were present in approximately equimolar amounts. Co elution of these proteins was also observed when the nuclear extract was fractionated by gel filtration, indicating that the NER proteins were associated in a complex with a molecular mass of >1000 kDa prior to affinity chromatography. The affinity purified NER complex catalyzed the incision of UV-irradiated DNA in an ATP-dependent reaction. We conclude that active high molecular weight complexes of NER proteins exist in undamaged yeast cells. PMID- 9852080 TI - Specific Cu2+-catalyzed oxidative cleavage of Na,K-ATPase at the extracellular surface. AB - This paper describes specific Cu2+-catalyzed oxidative cleavage of alpha and beta subunits of Na,K-ATPase at the extracellular surface. Incubation of right side out renal microsomal vesicles with Cu2+ ions, ascorbate, and H2O2 produces two major cleavages of the alpha subunit within the extracellular loop between trans membrane segments M7 and M8 and L7/8. Minor cleavages are also detected in loops L9/10 and L5/6. In the beta subunit two cleavages are detected, one before the first S-S bridge and the other between the second and third S-S bridges. Na,K ATPase and Rb+ occlusion are inactivated after incubation with Cu2+/ascorbate/H2O2. These observations are suggestive of a site-specific mechanism involving cleavage of peptide bonds close to a bound Cu2+ ion. This mechanism allows several inferences on subunit interactions and spatial organization. The two cleavage sites in L7/8 of the alpha subunit and two cleavage sites of the beta subunit identify interacting segments of the subunits. L7/8 is also close to L9/10 and to cation occlusion sites. Comparison of the locations of Cu2+-catalyzed cleavages with Fe2+-catalyzed cleavages (Goldshleger, R., and Karlish, S. J. D. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 9596-9601) suggests division of the membrane sector into two domains comprising M1-M6 and M7 M10/Mbeta, respectively. PMID- 9852081 TI - Highly cooperative homodimerization is a conserved property of neural POU proteins. AB - POU-domain proteins have been shown to play important roles in the development of the nervous, endocrine, and immune systems. However, the distinctive DNA recognition properties of the six major POU subclasses have not been well defined. Here, we have used random oligonucleotide selection and competitive binding assays to determine the optimal DNA recognition elements for the POU-III and POU-VI protein classes, represented by Brn-2 and Brn-5, respectively. The optimal Brn-5 consensus binding sequence GCATAA(T/A)TTAT strongly resembles that previously determined for the POU-IV (Brn-3) class, whereas Brn-2 exhibits highest affinity for non-octamer sites of the form ATG(A/C)AT(A/T)0-2ATTNAT and for octamer sites that contain a full associated heptamer sequence. Brn-2, Brn 3.0, and their invertebrate homologues all exhibit highly cooperative homodimerization on the Brn-2 consensus sequence, demonstrating that cooperative dimerization is a general property of these neural POU proteins. However, modified sites to which Brn-2 binds only as a monomer mediate the transcriptional effects of Brn-2 better than the consensus sequence, demonstrating that dimerization on these sites diminishes the transactivation ability of the protein. Together with the findings of our prior studies these data greatly facilitate the identification of functional POU recognition elements in the regulatory regions of neural genes. PMID- 9852082 TI - Heterologous expression of the Rhodobacter capsulatus BchI, -D, and -H genes that encode magnesium chelatase subunits and characterization of the reconstituted enzyme. AB - Magnesium chelatase inserts Mg2+ into protoporphyrin IX in the chlorophyll and bacteriochlorophyll biosynthetic pathways. In photosynthetic bacteria, the products of three genes, bchI, bchD, and bchH, are required for magnesium chelatase activity. These genes from Rhodobacter capsulatus were cloned separately into expression plasmids pET3a and pET15b. The pET15b constructs produced NH2-terminally His6-tagged proteins. All proteins were highly expressed and were purified to near homogeneity. The BchI and BchH proteins were soluble. BchD proteins were insoluble, inactive inclusion bodies that were renatured by rapid dilution from 6 M urea. The presence of BchI in the solution into which the urea solution of BchD was diluted increased the yield of active BchD. A molar ratio of 1 BchI:1 BchD was sufficient for maximum renaturation of BchD. All of the proteins were active in the magnesium chelatase assay except His-tagged BchI, which was inactive and inhibited in incubations containing non-His-tagged BchI. Expressed BchH proteins contained tightly bound protoporphyrin IX, and they were susceptible to inactivation by light. Maximum magnesium chelatase activity per mol of BchD occurred at a stoichiometry of 4 BchI:1 BchD. The optimum reaction pH was 8.0. The reaction exhibited Michaelis-Menten kinetics with respect to protoporphyrin IX and BchH. PMID- 9852083 TI - Assembly of a ternary complex by the predicted minimal coiled-coil-forming domains of syntaxin, SNAP-25, and synaptobrevin. A circular dichroism study. AB - The assembly of target (t-SNARE) and vesicle-associated SNAP receptor (v-SNARE) proteins is a critical step for the docking of synaptic vesicles to the plasma membrane. Syntaxin-1A, SNAP-25, and synaptobrevin-2 (also known as vesicle associated membrane protein, or VAMP-2) bind to each other with high affinity, and their binding regions are predicted to form a trimeric coiled-coil. Here, we have designed three peptides, which correspond to sequences located in the syntaxin-1A H3 domain, the C-terminal domain of SNAP-25, and a conserved central domain of synaptobrevin-2, that exhibit a high propensity to form a minimal trimeric coiled-coil. The peptides were synthesized by solid phase methods, and their interactions were studied by CD spectroscopy. In aqueous solution, the peptides were unstructured and showed no interactions with each other. In contrast, upon the addition of moderate amounts of trifluoroethanol (30%), the peptides adopted an alpha-helical structure and displayed both homomeric and heteromeric interactions. The interactions observed in ternary mixtures induce a stabilization of peptide structure that is greater than that predicted from individual binary interactions, suggesting the formation of a higher order structure compatible with the assembly of a trimeric coiled-coil. PMID- 9852084 TI - Characterization of FEN-1 from Xenopus laevis. cDNA cloning and role in DNA metabolism. AB - cDNAs for the Xenopus laevis homologue of the endo/exonuclease FEN-1 (DNase IV) have been cloned using a polymerase chain reaction strategy. Products were obtained from two nonallelic Xenopus genes (xFEN-1a and xFEN-1b) that differ from each other by 4.5% in amino acid sequence. Both are 80% identical to mammalian FEN-1 proteins and 55% identical to the yeast homologues. When expressed in Escherichia coli, the Xenopus enzymes showed flap endonuclease activity, a unique feature of this class of nucleases. In addition, expression from the Xenopus cDNAs complemented the temperature and methyl methanesulfonate sensitivity of a yeast rad27 deletion, which eliminates the endogenous FEN-1 gene product. Antiserum raised against xFEN-1 was used to show that the protein accumulates during the middle and late stages of oogenesis, in parallel with other DNA metabolic activities, and that it is localized to the oocyte nucleus. Flap endonuclease activity was demonstrated in oocyte nuclear extracts, and this was inhibited by the anti-xFEN-1 antiserum. The antiserum did not inhibit the major oocyte 5' --> 3' exonuclease activity. DNA synthesis in oocyte extracts was blocked by the antiserum, and the nature of this inhibition suggests that xFEN-1 may be part of a large complex of replication factors. Chromatographic evidence was obtained for the existence of a complex that forms during DNA synthesis and includes proliferating cell nuclear antigen in addition to xFEN-1. These observations support a critical role for xFEN-1 in DNA replication, but indicate that another enzyme must be responsible for the exonuclease function required for homologous recombination in Xenopus oocytes. PMID- 9852085 TI - NMR identification of an acyl-adenylate intermediate in the aryl-aldehyde oxidoreductase catalyzed reaction. AB - A new one-pot synthesis was designed to prepare benzoyl-AMP under anhydrous conditions in N,N-dimethylformamide. Reaction of benzoic acid with N,N' carbonyldiimidazole and subsequently with 5'-adenosyl monophosphate gave the mixed anhydride in 76% isolated yield. The structure of benzoyl-AMP was confirmed by mass spectroscopy and 1H-, 31P-, and 13C-NMR. The purity of the preparation was greater than 98% as indicated by 31P- and 13C-NMR. Purified aryl-aldehyde oxidoreductase was incubated in NMR tubes together with either carboxy-13C benzoyl-AMP or carboxy-13C-benzoic acid to demonstrate that benzoyl-AMP is an active intermediate during the enzymatic reduction of benzoic acid to benzaldehyde. PMID- 9852086 TI - Mutational analysis of Tyr-318 within the non-nucleoside reverse transcriptase inhibitor binding pocket of human immunodeficiency virus type I reverse transcriptase. AB - The highly conserved Tyr-318 is part of the non-nucleoside reverse transcriptase inhibitor (NNRTI)-specific lipophilic pocket of human immunodeficiency virus type I reverse transcriptase (RT) and makes contact within 4 A with the NNRTIs in all reported RT/NNRTI complexes. Using site-directed mutagenesis, six mutant RTs were constructed bearing the mutations Y318H, Y318K, Y318L, Y318C, Y318W, and Y318F. We found that only the Y318W and Y318F mutant RTs retained substantial RT activity, whereas the catalytic activities of the Y318K, Y318C, Y318H, and Y318L RT mutants were less than 5% of the wild-type activity. The Y318F mutant RT retained substantial sensitivity to the majority of NNRTIs tested, whereas the Y318W mutant RT showed varying degrees of resistance to NNRTIs. Subunit-specific site-directed mutagenesis revealed that there was no difference in the catalytic activity or resistance/sensitivity spectrum toward NNRTIs regardless of whether the Tyr-318 mutation was introduced in both subunits or only in the p66 subunit of RT. Recombinant viruses harboring the Y318F or Y318W mutation in the RT showed a similar resistance/sensitivity pattern to NNRTIs as their corresponding 318 mutant recombinant RTs. Our findings stress a functional or structural role for Tyr-318 in wild-type RT and argue for the design of novel NNRTIs that interact more closely with this amino acid in the NNRTI-specific pocket of human immunodeficiency virus type I RT. PMID- 9852087 TI - Mutations in the AF-2/hormone-binding domain of the chimeric activator GAL4.estrogen receptor.VP16 inhibit hormone-dependent transcriptional activation and chromatin remodeling in yeast. AB - GAL4.estrogen receptor.VP16 (GAL4.ER.VP16), which contains the GAL4 DNA-binding domain, the human ER hormone binding (AF-2) domain, and the VP16 activation domain, functions as a hormone-dependent transcriptional activator in yeast (Louvion, J.-F., Havaux-Copf, B., and Picard, D. (1993) Gene (Amst.) 131, 129 134). Previously, we showed that this activator can remodel chromatin in yeast in a hormone-dependent manner. In this work, we show that a weakened VP16 activation domain in GAL4.ER.VP16 still allows hormone-dependent chromatin remodeling, but mutations in the AF-2 domain that abolish activity in the native ER also eliminate the ability of GAL4.ER.VP16 to activate transcription and to remodel chromatin. These findings suggest that an important role of the AF-2 domain in the native ER is to mask the activation potential of the AF-1 activation domain in the unliganded state; upon ligand activation, a conformational change releases AF-2-mediated repression and transcriptional activation ensues. We also show that the AF-2 domain, although inactive at simple promoters on its own in yeast, can enhance transcription by the MCM1 activator in hormone-dependent manner, consistent with its having a role in activation as well as repression in the native ER. PMID- 9852088 TI - 1.85-A resolution crystal structure of human ornithine transcarbamoylase complexed with N-phosphonacetyl-L-ornithine. Catalytic mechanism and correlation with inherited deficiency. AB - The crystal structure of human ornithine transcarbamoylase complexed with the bisubstrate analog N-phosphonacetyl-L-ornithine has been solved at 1.85-A resolution by molecular replacement. Deleterious mutations produce clinical hyperammonia that, if untreated, results in neurological symptoms or death (ornithine transcarbamylase deficiency). The holoenzyme is trimeric, and as in other transcarbamoylases, each subunit contains an N-terminal domain that binds carbamoyl phosphate and a C-terminal domain that binds L-ornithine. The active site is located in the cleft between domains and contains additional residues from an adjacent subunit. Binding of N-phosphonacetyl-L-ornithine promotes domain closure. The resolution of the structure enables the role of active site residues in the catalytic mechanism to be critically examined. The side chain of Cys-303 is positioned so as to be able to interact with the delta-amino group of L ornithine which attacks the carbonyl carbon of carbamoyl phosphate in the enzyme catalyzed reaction. This sulfhydryl group forms a charge relay system with Asp 263 and the alpha-amino group of L-ornithine, instead of with His-302 and Glu 310, as previously proposed. In common with other ureotelic ornithine transcarbamoylases, the human enzyme lacks a loop of approximately 20 residues between helix H10 and beta-strand B10 which is present in prokaryotic ornithine transcarbamoylases but has a C-terminal extension of 10 residues that interacts with the body of the protein but is exposed. The sequence of this C-terminal extension is homologous to an interhelical loop found in several membrane proteins, including mitochondrial transport proteins, suggesting a possible mode of interaction with the inner mitochondrial membrane. PMID- 9852089 TI - Escherichia coli DnaA protein. The N-terminal domain and loading of DnaB helicase at the E. coli chromosomal origin. AB - Initiation of DNA replication at the Escherichia coli chromosomal origin occurs through an ordered series of events that depends first on the binding of DnaA protein, the replication initiator, to DnaA box sequences followed by unwinding of an AT-rich region. A step that follows is the binding of DnaB helicase at oriC so that it is properly positioned at each replication fork. We show that DnaA protein actively mediates the entry of DnaB at oriC. One region (amino acids 111 148) transiently binds to DnaB as determined by surface plasmon resonance. A second functional domain, possibly involving formation of a unique nucleoprotein structure, promotes the stable binding of DnaB during the initiation process and is inactivated in forming an intermediate termed the prepriming complex by removal of the N-terminal 62 residues. Based on similarities in the replication process between prokaryotes and eukaryotes, these results suggest that a similar mechanism may load the eukaryotic replicative helicase. PMID- 9852090 TI - Adenovirus-mediated gene transfer of cGMP-dependent protein kinase increases the sensitivity of cultured vascular smooth muscle cells to the antiproliferative and pro-apoptotic effects of nitric oxide/cGMP. AB - Studies in vitro have underestimated the importance of cGMP-dependent protein kinase (PKG) in the modulation of vascular smooth muscle cell (SMC) proliferation and apoptosis in vivo. This is attributable, in part, to a rapid decline in PKG levels as vascular SMC are passaged in culture. We used a recombinant adenovirus encoding PKG (Ad.PKG) to augment kinase activity in cultured rat pulmonary artery SMC (RPaSMC). Incubation of Ad. PKG-infected RPaSMC (multiplicity of infection = 200) with 8-Br-cGMP decreased serum-stimulated DNA synthesis by 85% and cell proliferation at day 5 by 74%. The effect of 8-Br-cGMP on DNA synthesis in Ad.PKG infected RPaSMC was blocked by KT5823 (PKG inhibitor), but not by KT5720 (cAMP dependent protein kinase inhibitor). A nitric oxide (NO) donor compound, S nitrosoglutathione, at concentrations as low as 100 nM, inhibited DNA synthesis in Ad. PKG-infected RPaSMC, but not in uninfected cells or in cells infected with a control adenovirus. In addition, 8-Br-cGMP and S-nitrosoglutathione induced apoptosis in serum-deprived RPaSMC infected with Ad.PKG, but not in uninfected cells or in cells infected with a control adenovirus. These results demonstrate that modulation of PKG levels in vascular SMC can alter the sensitivity of these cells to NO and cGMP. Moreover, these observations suggest an important role for PKG in the regulation of vascular SMC proliferation and apoptosis by NO and cGMP. PMID- 9852091 TI - Bcl-2 prevents caspase-independent cell death. AB - Nitric oxide (NO) is implicated in apoptosis and has both cytotoxic and cytoprotective effects. Exogenous NO induced the death of PC12 and HeLa cells via a process showing features of both apoptosis and necrosis, with chromatin condensation, nuclear compaction, and mitochondrial swelling. Activation of caspases was not observed during NO-induced cell death. In addition, cell death was not inhibited by peptide caspase inhibitors or by expression of p35, a baculovirus-encoded caspase inhibitor, indicating that NO-induced cell death was independent of caspases. NO-induced cell death was enhanced by Bax expression in a caspase-independent manner and prevented by the anti-cell death protein Bcl-2. Although Bcl-2 has previously been shown to prevent cell death by inhibiting caspase activation, these results indicate that it can also prevent cell death via a caspase-independent mechanism. PMID- 9852092 TI - Granzyme B mimics apical caspases. Description of a unified pathway for trans activation of executioner caspase-3 and -7. AB - Granzyme B (GrB) is predicted to trigger apoptosis by activating preferred caspases, but the zymogens that are directly processed by the granzyme and the requirements for these interactions remain unclarified. We examined this dilemma by comparing the kinetics and pattern of GrB-mediated activation of the executioner caspase-7 in vitro and in vivo. GrB rapidly activates procaspase-7 in vitro by cleaving between the large and small subunits leaving the propeptide intact. During GrB-mediated apoptosis, the caspase-7 propeptide is removed and cleavage occurs between the subunits. Strikingly, caspase-7 is unprocessed in caspase-3-deficient MCF-7 cells exposed to GrB but is rapidly activated when the cells are solubilized. Transfection with caspase-3 restores the removal of the caspase-7 propeptide and the capacity of GrB to subsequently activate the caspase. The data suggest that GrB activates caspase-3, which then removes the propeptide of caspase-7 allowing activation by GrB. Thus GrB initiates the death pathway by processing the accessible caspase-3, and the caspase-7 propeptide regulates trans-activation of the zymogen by granzyme. As a consequence, two proteases, caspase-3 and GrB, are required to activate procaspase-7. PMID- 9852093 TI - Mechanism of allosteric regulation of the rod cGMP phosphodiesterase activity by the helical domain of transducin alpha subunit. AB - The G protein alpha subunit (Galpha) is composed of two distinct folding domains: a GTP-binding Ras-like domain and an alpha helical domain (HD). We have recently reported that the helical domain (HDt) of the vertebrate visual transducin alpha subunit (Galphat) synergizes activation of retinal cyclic GMP phosphodiesterase (PDE) by activated Galphat (Liu, W., and Northup, J. K., (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 12878-12883). Here, we examine the molecular basis for this HD based signaling regulation, and we provide a new model for the activation of the target effector. The HD proteins derived from visual transducin or taste gustducin alpha subunits, but no other Galpha HD proteins, each attenuate the PDE catalytic core (Palphabeta) and synergize Galphat stimulation of the holoPDE (Palphabetagamma2) with similar apparent affinities. The data from studies of both HDt-mediated attenuation and stimulation indicate that the HDt and the PDE inhibitory subunit (Pgamma) interact with PDE at independent sites and that Palphabeta contains the binding sites for HD. The saturation of both processes by HDt displays positive cooperativity with Hill coefficients of 1.5 for the attenuation of Palphabeta activity and 2.1 for synergism of holoPDE activation. Our data suggest the that Galphat-HDt regulates PDE by allosterically decreasing the affinity of Palphabeta for Pgamma and thus simultaneously facilitating the interaction of the activated Galphat-Ras-like domain with Pgamma. Thus, we propose a new model for the high efficiency of PDE activation as well as deactivation, and, overall, a novel mechanism for controlling fidelity, sensitivity, and efficacy of G protein signaling. PMID- 9852094 TI - Sorbitol dehydrogenase of Drosophila. Gene, protein, and expression data show a two-gene system. AB - The Drosophila melanogaster sorbitol dehydrogenase (SDH) is characterized as a two-enzyme system of the medium chain dehydrogenase/reductase family (MDR). The SDH-1 enzyme has an enzymology with Km and kcat values an order of magnitude higher than those for the human enzyme but with a similar kcat/Km ratio. It is a tetramer with identical subunits of approximately 38 kDa. At the genomic level, two genes, Sdh-1 and Sdh-2, have a single transcriptional start site and no functional TATA box. Expression is greater in larvae and adults than in pupae, where it is very low. At all three stages, Sdh-1 constitutes the major transcript. Sdh-1 and Sdh-2 genes were located at positions 84E-F and 86D in polytene chromosomes. The deduced amino acid sequences of the two genes show 90% residue identity. Evaluation of the sequence and modeling of the structure toward that of class I alcohol dehydrogenase (ADH) show altered loop and gap arrangements as in mammalian SDH and establishes that SDH, despite gene multiplicity and larger variability than the "constant" ADH of class III, is an enzyme conserved over wide ranges. PMID- 9852096 TI - Structural and thermodynamic responses of mutations at a Ca2+ binding site engineered into human lysozyme. AB - Structural determinants of Ca2+ binding sites within proteins typically comprise several acidic residues in appropriate juxtaposition. Three residues (Ala-83, Gln 86, and Ala-92) in human lysozyme are characteristically mutated to Lys, Asp, and Asp, respectively, in natural Ca2+ binding lysozymes and alpha-lactalbumins. The effects of these mutations on the stability and Ca2+ binding properties of human lysozyme were investigated using calorimetry and were interpreted with crystal structures. The double mutant, in which Glu-86 and Ala-92 were replaced with Asp, clearly showed Ca2+ binding affinity, whereas neither point mutant showed Ca2+ affinity, indicating that both residues are essential. The further mutation of Ala-83 --> Lys did not affect the Ca2+ binding of the double mutant. The point mutations Ala-83 --> Lys and Glu-86 --> Asp did not affect the stability, whereas the mutation Ala-92 --> Asp was about 1.3 kcal/mol less stable. Structural analyses showed that both Asp-86 and Lys-83 were exposed to solvent. Side chains of Asp-86 and Asp-91 were rotated in opposite directions about chi1 angle, as if to reduce the electrostatic repulsion. The charged amino acids at the Ca2+ binding site did not significantly affect stability of the protein, possibly because of the local conformational change of the side chains. PMID- 9852095 TI - E3-ubiquitin ligase/E6-AP links multicopy maintenance protein 7 to the ubiquitination pathway by a novel motif, the L2G box. AB - Ubiquitin ligases are generally assumed to play a major role in substrate recognition and thus provide specificity to a particular ubiquitin modification system. The multicopy maintenance protein (Mcm) 7 subunit of the replication licensing factor-M was identified as a substrate of the E3-ubiquitin ligase/E6-AP by its interaction with human papillomavirus-18E6. Mcm7 is ubiquitinated in vivo in both an E6-AP-dependent and -independent manner. E6-AP functions in these reactions independently of the viral oncogene E6. We show that recognition of Mcm7 by E6-AP is mediated by a homotypic interaction motif present in both proteins, called the L2G box. These findings served as the basis for the definition of substrate specificity for E6-AP. A small cluster of proteins whose function is intimately associated with the control of cell growth and/or proliferation contains the L2G box and is thereby implicated in an E6-AP and, by default, HPV-E6-dependent ubiquitination pathway. PMID- 9852097 TI - cDNA cloning of mouse and human cholesterol 25-hydroxylases, polytopic membrane proteins that synthesize a potent oxysterol regulator of lipid metabolism. AB - Oxysterols regulate the expression of genes involved in cholesterol and lipid metabolism and serve as intermediates in cholesterol catabolism. Among the most potent of regulatory oxysterols is 25-hydroxycholesterol, whose biosynthetic enzyme has not yet been isolated. Here, we report the cloning of cholesterol 25 hydroxylase cDNAs from the mouse and human. The encoded enzymes are polytopic membrane proteins of 298 and 272 amino acids, respectively, which contain clusters of histidine residues that are essential for catalytic activity. Unlike most other sterol hydroxylases, cholesterol 25-hydroxylase is not a cytochrome P450, but rather it is a member of a small family of enzymes that utilize diiron cofactors to catalyze the hydroxylation of hydrophobic substrates. The cholesterol 25-hydroxylase gene lacks introns, and in the human it is located on chromosome 10q23. The murine gene is expressed at low levels in multiple tissues. Expression of cholesterol 25-hydroxylase in transfected cells reduces the biosynthesis of cholesterol from acetate and suppresses the cleavage of sterol regulatory element binding protein-1 and -2. The data suggest that cholesterol 25 hydroxylase has the capacity to play an important role in regulating lipid metabolism by synthesizing a co-repressor that blocks sterol regulatory element binding protein processing and ultimately leads to inhibition of gene transcription. PMID- 9852098 TI - Evidence for a salt bridge between transmembrane segments 5 and 6 of the yeast plasma-membrane H+-ATPase. AB - The plasma-membrane H+-ATPase of Saccharomyces cerevisiae, which belongs to the P2 subgroup of cation-transporting ATPases, is encoded by the PMA1 gene and functions physiologically to pump protons out of the cell. This study has focused on hydrophobic transmembrane segments M5 and M6 of the H+-ATPase. In particular, a conserved aspartate residue near the middle of M6 has been found to play a critical role in the structure and biogenesis of the ATPase. Site-directed mutants in which Asp-730 was replaced by an uncharged residue (Asn or Val) were abnormally sensitive to trypsin, consistent with the idea that the proteins were poorly folded, and immunofluorescence confocal microscopy showed them to be arrested in the endoplasmic reticulum. Similar defects are known to occur when either Arg-695 or His-701 in M5 is replaced by a neutral residue (Dutra, M. B., Ambesi, A., and Slayman, C. W. (1998) J. Biol. Chem. 273, 17411-17417). To search for possible charge-charge interactions between Asp-730 and Arg-695 or His-701, double mutants were constructed in which positively and negatively charged residues were swapped or eliminated. Strikingly, two of the double mutants (R695D/D730R and R695A/D730A) regained the capacity for normal biogenesis and displayed near-normal rates of ATP hydrolysis and ATP-dependent H+ pumping. These results demonstrate that neither Arg-695 nor Asp-730 is required for enzymatic activity or proton transport, but suggest that there is a salt bridge between the two residues, linking M5 and M6 of the 100-kDa polypeptide. PMID- 9852099 TI - Release of the neuregulin functional polypeptide requires its cytoplasmic tail. AB - Based on both in vivo and in vitro studies, we have shown previously that the intracellular domain of a membrane-bound isoform of the growth factor, neuregulin, regulates proteolytic release of its extracellular domain ErbB receptor-activating ligand. To investigate the mechanism(s) involved in this regulation, a series of intracellular domain mutants were constructed and tested for susceptibility to proteolytic processing after transient transfection in COS 7 cells. These studies revealed that regulation of extracellular domain cleavage by the intracellular domain is sequence-specific and involves three distinct 30 60-residue segments. The presence of any two of these three segments is both necessary and sufficient for proteolytic processing, and resistance to proteolysis is not due to an alteration in cellular localization or transport. Evidence was also obtained that regulation of extracellular domain processing involves initial intracellular domain dimerization. Thus, with expression of a construct encoding only the intracellular domain, dimerization could be demonstrated in cross-linking experiments. Furthermore, resistance to proteolytic processing of a construct lacking a large portion of the intracellular domain was rescued with a chimera, in which the intracellular domain was replaced with a spontaneously dimerizing Fc fragment. Taken together these studies indicate that intracellular domain interactions are critically involved in the spacial and temporal control of growth and development by membrane-bound neuregulin isoforms. PMID- 9852100 TI - The cellular localization of the murine serine/arginine-rich protein kinase CLK2 is regulated by serine 141 autophosphorylation. AB - Pre-mRNA splicing is catalyzed by a multitude of proteins including serine/arginine-rich (SR) proteins, which are thought to play a crucial role in the formation of spliceosomes and in the regulation of alternative splicing. SR proteins are highly phosphorylated, and their kinases are believed to regulate the recruitment of SR proteins from nuclear storage compartments known as speckles. Recently, a family of autophosphorylating kinases termed CLK (CDC2/CDC28-like kinases) was shown to phosphorylate SR proteins and to influence alternative splicing in overexpression systems. Here we used endogenous CLK2 protein to demonstrate that it displays different biochemical characteristics compared with its overexpressed protein and that it is differentially phosphorylated in vivo. Furthermore, CLK2 changed its nuclear localization upon treatment with the kinase inhibitor 5, 6-dichloro-1-beta-D ribofuranosylbenzimidazole. We have also identified a CLK2 autophosphorylation site, which is highly conserved among all CLK proteins, and we show by site directed mutagenesis that its phosphorylation influences the subnuclear localization of CLK2. Our data suggest that CLK2 localization and possibly activity are influenced by a balance of CLK2 autophosphorylation and the regulation by CLK2 kinases and phosphatases. PMID- 9852101 TI - Lactosylceramide mediates tumor necrosis factor-alpha-induced intercellular adhesion molecule-1 (ICAM-1) expression and the adhesion of neutrophil in human umbilical vein endothelial cells. AB - The endothelial expression of adhesion molecules by proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) has been suggested to contribute to the initiation of atherosclerotic plaque formation. Since lactosylceramide (LacCer) accumulates in large quantities in human atherosclerotic plaque, we have explored its role in TNF-alpha-induced expression of intercellular adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells and their consequent adhesion to polymorphonuclear leukocytes (PMNs). We found that TNF alpha increased LacCer synthesis by way of stimulating the activity of UDP galactose:glucosylceramide beta(1-->4)-galactosyltransferase in a time-dependent fashion. The TNF-alpha-induced expression of ICAM-1 was abrogated by D-1-phenyl-2 decanoylamino-3-morpholino-1-propanol (D-PDMP), an inhibitor of UDP galactose:glucosylceramide beta(1-->4)-galactosyltransferase. However, the addition of LacCer reversed the D-PDMP effect on TNF-alpha-induced ICAM-1 expression in human umbilical vein endothelial cells. Northern hybridization analysis of mRNA levels and enzyme-linked immunosorbent assays revealed that LacCer (5 microM) specifically stimulated ICAM-1 at both the transcriptional and translational levels. This was accompanied by the adhesion of PMNs, which was visualized by confocal microscopy. Further studies revealed that LacCer stimulated the endogenous generation of superoxide radicals (O-2) about 5-fold compared with the control by specifically activating plasma membrane-associated NADPH-dependent oxidase. This phenomenon was blocked by the antioxidant N-acetyl L-cysteine, pyrrolidine dithiocarbamate, and the NADPH oxidase inhibitor, diphenylene iodonium. Overexpression of endogeneous CuZn-superoxide dismutase via an adenoviral vector carrying cDNA for CuZn-superoxide dismutase, also inhibited LacCer-induced ICAM-1 expression in endothelial cells. In sum, our findings suggest that LacCer may play the role of a lipid second messenger in TNF-alpha induced pathogenesis by activating an oxidant-sensitive transcriptional pathway that leads to the adhesion of PMNs to endothelial cells. PMID- 9852102 TI - Evolution, organization, and expression of alpha-tubulin genes in the antarctic fish Notothenia coriiceps. Adaptive expansion of a gene family by recent gene duplication, inversion, and divergence. AB - To assess the organization and expression of tubulin genes in ectothermic vertebrates, we have chosen the Antarctic yellowbelly rockcod, Notothenia coriiceps, as a model system. The genome of N. coriiceps contains approximately 15 distinct DNA fragments complementary to alpha-tubulin cDNA probes, which suggests that the alpha-tubulins of this cold-adapted fish are encoded by a substantial multigene family. From an N. coriiceps testicular DNA library, we isolated a 13.8-kilobase pair genomic clone that contains a tightly linked cluster of three alpha-tubulin genes, designated NcGTbalphaa, NcGTbalphab, and NcGTbalphac. Two of these genes, NcGTbalphaa and NcGTbalphab, are linked in head to-head (5' to 5') orientation with approximately 500 bp separating their start codons, whereas NcGTbalphaa and NcGTbalphac are linked tail-to-tail (3' to 3') with approximately 2.5 kilobase pairs between their stop codons. The exons, introns, and untranslated regions of the three alpha-tubulin genes are strikingly similar in sequence, and the intergenic region between the alphaa and alphab genes is significantly palindromic. Thus, this cluster probably evolved by duplication, inversion, and divergence of a common ancestral alpha-tubulin gene. Expression of the NcGTbalphac gene is cosmopolitan, with its mRNA most abundant in hematopoietic, neural, and testicular tissues, whereas NcGTbalphaa and NcGTbalphab transcripts accumulate primarily in brain. The differential expression of the three genes is consistent with distinct suites of putative promoter and enhancer elements. We propose that cold adaptation of the microtubule system of Antarctic fishes is based in part on expansion of the alpha and beta-tubulin gene families to ensure efficient synthesis of tubulin polypeptides. PMID- 9852103 TI - The box-1 region of the leukemia inhibitory factor receptor alpha-chain cytoplasmic domain is sufficient for hemopoietic cell proliferation and differentiation. AB - Leukemia inhibitory factor (LIF) is a pleiotropic cytokine that acts on a variety of cell types and regulates cell proliferation and differentiation. The functional receptor for LIF is composed of LIFR alpha-chain (LIFRalpha) and gp130 both of which are shared in the functional receptors for oncostatin M, ciliary neurotrophic factor, and cardiotrophin-1. By using stable transfection of wild type or cytoplasmic deletion mutants of LIFRalpha together with full-length gp130 into Ba/F3 cells, we found that cells expressing gp130 and an extensively deleted mutant LIFRalpha containing only the box-1 region were capable of proliferating in response to LIF, although LIF-dependent long term growth of these cells was seriously impaired. Using a similar strategy to generate WEHI-3BD+ cells expressing gp130 and wild-type or truncation mutants of LIFRalpha, studies revealed that the box-1 region of the LIFRalpha was also sufficient for LIF dependent induction of different aspects of differentiation, including up regulation of macrophage surface marker expression, morphological change, and cell migration in agar culture. However, the C-terminal region of the LIFRalpha, although not essential for intracellular signaling, was important for efficient receptor-mediated ligand internalization. In summary, the membrane-proximal box-1 region plays a dominant role in LIF-induced signal transduction of both proliferation and differentiation. PMID- 9852104 TI - Identification of tethering domains for protein kinase A type Ialpha regulatory subunits on sperm fibrous sheath protein FSC1. AB - The fibrous sheath is a unique cytoskeletal structure in the sperm flagellum believed to modulate sperm motility. FSC1 is the major structural protein of the fibrous sheath. The yeast two-hybrid system was used to identify other proteins that contribute to the structure of the fibrous sheath or participate in sperm motility. When FSC1 was used as the bait to screen a mouse testis cDNA library, two clones were isolated encoding the type Ialpha regulatory subunit (RIalpha) of cAMP-dependent protein kinase. Deletion analysis using the yeast two-hybrid system and in vitro binding assays with glutathione S-transferase-FSC1 fusion proteins identified two RIalpha tethering domains on FSC1. A domain located at residues 219-232 (termed domain A) corresponds to the reported tethering domain for a type II regulatory subunit (RII) of cAMP-dependent protein kinase, indicating that this binding domain has dual specificity to RI and RII. Another RIalpha tethering site (termed domain B) at residues 335-344 shows specific binding of RIalpha and had no significant sequence homology with known RII tethering domains. However, helical wheel projection analysis indicates that domain B is likely to form an amphipathic helix, the secondary structure of RII tethering domains of protein kinase A anchoring proteins. This was supported by the finding that site-directed mutagenesis to disrupt the amphipathic helix eliminated RIalpha binding. This is apparently the first report of an RIalpha specific protein kinase A anchoring protein tethering domain. PMID- 9852105 TI - A Mycoplasma fermentans-derived synthetic lipopeptide induces AP-1 and NF-kappaB activity and cytokine secretion in macrophages via the activation of mitogen activated protein kinase pathways. AB - Mycoplasma lipoproteins have been demonstrated to stimulate monocytic cells and induce proinflammatory cytokine secretion. In this paper, we show that a synthetic analog of the Mycoplasma fermentans membrane-associated lipopeptide macrophage-activating lipopeptide-2 (MALP-2) induces mRNA synthesis and protein secretion of interleukin-1beta and tumor necrosis factor-alpha in human monocytes/macrophages and the murine macrophage cell line RAW 264.7, whereas the nonlipidated counterpart lacks this effect, underscoring the importance of protein acylation for cell activation. Synthetic MALP-2 (sMALP-2) induced the activation of MAPK family members extracellular signal regulated kinases 1 and 2, c-Jun NH2-terminal kinase, and p38 and induced NF-kappaB and AP-1 transactivation in macrophages. Whereas the specific p38 inhibitor SB203580 abrogated both cytokine synthesis and NF-kappaB and AP-1 transactivation in response to MALP-2, the selective MAPK/extracellular signal-regulated kinase-1 inhibitor PD-98059 decreased interleukin-1beta and tumor necrosis factor-alpha production in response to sMALP-2 without affecting the transactivation of NF-kappaB or AP-1. These results indicate that activation of MAPKs by sMALP-2 is a crucial event leading to the expression of proinflammatory cytokines. Our findings demonstrate that the synthetic analog of MALP-2 reproduces the macrophage stimulation activity found in different fractions of mycoplasmas. Given that MALP-2 has been recently shown to be expressed at the surface of M. fermentans as a molecular entity, sMALP-2 constitutes a valuable surrogate for investigating immunomodulation by these microorganisms and evaluating the role that this activity plays in the development of inflammatory diseases associated with mycoplasma infections. PMID- 9852106 TI - Loss of Drs2p does not abolish transfer of fluorescence-labeled phospholipids across the plasma membrane of Saccharomyces cerevisiae. AB - The yeast DRS2 gene, which is required for growth at 23 degreesC or below, encodes a member of a P-type ATPase subgroup reported to transport aminophospholipids between the leaflets of the plasma membrane. Here, we evaluated the potential role of Drs2p in phospholipid transport. When examined by fluorescence microscopy, a drs2 null mutant showed no defect in the uptake or distribution of fluorescent-labeled 1-palmitoyl-2[6-(7-nitrobenz-2-oxa-1, 3 diazol-4-yl (NBD))aminocaproyl]phosphatidylserine) or 1-myristoyl-2[6-NBD aminocaproyl]phosphatidylethanolamine. Quantification of the amount of cell associated NBD fluorescence using flow cytometry indicated a significant decrease in the absence of Drs2p, but this decrease was not restricted to the aminophospholipids (phosphatidylserine and phosphatidylethanolamine) and was dependent on culture conditions. Furthermore, the absence of Drs2p had no effect on the amount of endogenous PE exposed to the outer leaflet of the plasma membrane as detected by labeling with trinitrobenzene sulfonic acid. The steady state pool of Drs2p, which was shown to reside predominantly in the plasma membrane, increased upon shift to low temperature or exposure to various divalent cations (Mn2+, Co2+, Ni2+, and Zn2+ but not Ca2+ or Mg2+), conditions that also inhibited the growth of a drs2 null mutant. The data presented here call into question the identification of Drs2p as the exclusive or major aminophospholipid translocase in yeast plasma membranes (Tang, X., Halleck, M. S., Schlegel, R. A., and Williamson, P. (1996) Science 272, 1495-1497). PMID- 9852107 TI - Human HtrA, an evolutionarily conserved serine protease identified as a differentially expressed gene product in osteoarthritic cartilage. AB - The human homologue of the Escherichia coli htrA gene product was identified by the differential display analysis of transcripts expressed in osteoarthritic cartilage. This transcript was identified previously as being repressed in SV40 transformed fibroblasts (Zumbrunn, J., and Trueb, B. (1996) FEBS Lett. 398, 187 192). Levels of HtrA mRNA were elevated approximately 7-fold in cartilage from individuals with osteoarthritis compared with nonarthritic controls. Differential expression of human HtrA protein was confirmed by an immunoblot analysis of cartilage extracts. Human HtrA protein expressed in heterologous systems was secreted and exhibited endoproteolytic activity, including autocatalytic cleavage. Conversion by mutagenesis of the putative active site serine 328 to alanine eliminated the enzymatic activity. Serine 328 was also found to be required for the formation of a stable complex with alpha1-antitrypsin. We have determined that the HtrA gene is highly conserved among mammalian species: the amino acid sequences encoded by HtrA cDNA clones from cow, rabbit, and guinea pig are 98% identical to human. In E. coli, a functional htrA gene product is required for cell survival after heat shock or oxidative stress; its role appears to be the degradation of denatured proteins. We propose that mammalian HtrA, with the addition of a new functionality during evolution, i.e. a mac25 homology domain, plays an important role in cell growth regulation. PMID- 9852108 TI - Identification of novel pancreas-specific regulatory sequences in the promoter region of human pancreatic secretory trypsin inhibitor gene. AB - The human pancreatic secretory trypsin inhibitor (PSTI) genes introduced into mice are specifically expressed in pancreas. The 1.0 kilobase pairs of PSTI 5' flanking sequence directed preferential expression of a linked reporter chloramphenicol acetyltransferase, which was active in a PSTI-expressing pancreatic cell line (AR42j) but not in a PSTI-nonexpressing fibroblast cell line (XC). Two positively acting elements were found, Region I (-161/-116) and Region II (-103/-74), as defined by transfection and binding assays with AR42j cells. Region II is sufficient for the pancreas-specific expression, but the presence of both Regions I and II is needed for the maximum activity. Sequence studies also revealed that these two elements differ from the previously identified recognition sequence for pancreas transcription factor 1 (PTF1). When the same set of experiments was done with XC cells, one negatively acting element was identified, Region IV (-154/-137). Interestingly, Regions I and IV share a core sequence (-149/-139), CAATCAATAAC. These results suggest that this novel element regulates the human PSTI gene expression positively in pancreatic cells but negatively in nonpancreatic cells. PMID- 9852109 TI - Regulation of rat pyruvate carboxylase gene expression by alternate promoters during development, in genetically obese rats and in insulin-secreting cells. Multiple transcripts with 5'-end heterogeneity modulate translation. AB - A previous study on the gene structure of rat pyruvate carboxylase revealed that two tissue-specific promoters are responsible for the production of multiple transcripts with 5'-end heterogeneity (Jitrapakdee, S., Booker, G. W., Cassady, A. I., and Wallace, J. C. (1997) J. Biol. Chem. 272, 20522-20530). Here we report transcription and translation regulation of pyruvate carboxylase (PC) expression during development and in genetically obese rats. The abundance of PC mRNAs was low in fetal liver but increased by 2-4-fold within 7 days after birth, concomitant with an 8-fold increase in the amount of immunoreactive PC and its activity and then decreased during the weaning period. Reverse transcriptase polymerase chain reaction analysis indicated that the proximal promoter was activated during the suckling period and reduced in activity at weaning. In genetically obese Zucker rats, adipose PC was 4-5-fold increased, concomitant with a 5-6-fold increase in mRNA level. Reverse transcriptase-polymerase chain reaction analysis also showed that the proximal promoter was activated in the hyperlipogenic condition. Conversely, transcription of the proximal promoter was not detectable in various liver cell lines, suggesting that this promoter was not functional under cell culture conditions. In rat pancreatic islets and insulinoma cells, only transcripts D and E, generated from the distal promoter of the PC gene, were expressed. Glucose increased PC transcripts from the distal promoter when the insulinoma cells were maintained in 10 mM glucose. We conclude that the proximal promoter of the rat PC gene plays a major role in gluconeogenesis and lipogenesis, whereas the distal promoter is necessary for anaplerosis. In vitro translation and in vivo polysome profile analysis indicated that transcripts C and E were translated with similar translational efficiencies that are substantially greater than that of transcript D, suggesting that 5'-untranslated regions play a role in translational control. PMID- 9852110 TI - Differential activity of the G protein beta5 gamma2 subunit at receptors and effectors. AB - The G protein beta5 subunit differs substantially in amino acid sequence from the other known beta subunits suggesting that beta gamma dimers containing this protein may play specialized roles in cell signaling. To examine the functional properties of the beta5 subunit, recombinant beta5 gamma2 dimers were purified from baculovirus-infected Sf9 insect cells using a strategy based on two affinity tags (hexahistidine and FLAG) engineered into the N terminus of the gamma2 subunit (gamma2HF). The function of the pure beta5 gamma2HF dimers was examined in three assays: activation of pure phospholipase C-beta in lipid vesicles; activation of recombinant, type II adenylyl cyclase expressed in Sf9 cell membranes; and coupling of alpha subunits to the endothelin B (ETB) and M1 muscarinic receptors. In each case, the efficacy of the beta5 gamma2HF dimer was compared with that of the beta1 gamma2HF dimer, which has demonstrated activity in these assays. The beta5 gamma2HF dimer activated phospholipase C-beta with a potency and efficacy similar to that of beta1 gamma2 or beta1 gamma2HF; however, it was markedly less effective than the beta1 gamma2HF or beta1 gamma2 dimer in its ability to activate type II adenylyl cyclase (EC50 of approximately 700 nM versus 25 nM). Both the beta5 gamma2HF and the beta1 gamma2HF dimers supported coupling of M1 muscarinic receptors to the Gq alpha subunit. The ETB receptor coupled effectively to both the Gi and Gq alpha subunits in the presence of the beta1 gamma2HF dimer. In contrast, the beta5 gamma2HF dimer only supported coupling of the Gq alpha subunits to the ETB receptor and did not support coupling of the Gi alpha subunit. These results suggest that the beta5 gamma2HF dimer binds selectively to Gq alpha subunits and does not activate the same set of effectors as dimers containing the beta1 subunit. Overall, the data support a specialized role for the beta5 subunit in cell signaling. PMID- 9852112 TI - Immunoaffinity purification and functional characterization of human transcription factor IIH and RNA polymerase II from clonal cell lines that conditionally express epitope-tagged subunits of the multiprotein complexes. AB - Purification of multiprotein complexes such as transcription factor (TF) IIH and RNA polymerase II (pol II) has been a tedious task by conventional chromatography. To facilitate the purification, we have developed an effective scheme that allows human TFIIH and pol II to be isolated from HeLa-derived cell lines that conditionally express the FLAG-tagged p62 subunit of human TFIIH and the RPB9 subunit of human pol II, respectively. An approximate 2000-fold enrichment of FLAG-tagged TFIIH and a 1000-fold enhancement of total pol II are achieved by a one-step immunoaffinity purification. The purified complexes are functional in mediating basal and activated transcription, regardless of whether TATA-binding protein or TFIID is used as the TATA-binding factor. Interestingly, repression of basal transcription by the positive cofactor PC4 is alleviated by increasing amounts of TFIID, TFIIH, and pol II holoenzyme, suggesting that phosphorylation of PC4 by these proteins may cause a conformational change in the structure of PC4 that allows for preinitiation complex formation and initiation of transcription. Furthermore, pol II complexes with different phosphorylation states on the carboxyl-terminal domain of the largest subunit are selectively purified from the inducible pol II cell line, making it possible to dissect the role of carboxyl-terminal domain phosphorylation in the transcription process in a highly defined in vitro transcription system. PMID- 9852111 TI - The p85 subunit of phosphatidylinositol 3-kinase associates with the Fc receptor gamma-chain and linker for activitor of T cells (LAT) in platelets stimulated by collagen and convulxin. AB - There is extensive evidence to show that phosphatidylinositol 3-kinase plays an important role in signaling by the immune family of receptors, which has recently been extended to include the platelet collagen receptor, glycoprotein VI. In this report we present two potential mechanisms for the regulation of this enzyme on stimulation of platelets by collagen. We show that on stimulation with collagen, the regulatory subunit of phosphatidylinositol 3-kinase associates with the tyrosine-phosphorylated form of the adapter protein linker for activator of T Cells (LAT) and the tyrosine-phosphorylated immunoreceptor tyrosine-based activation motif of the Fc receptor gamma-chain (a component of the collagen receptor complex that includes glycoprotein VI). The associations of the Fc receptor gamma-chain and LAT with p85 are rapid and supported by the Src-homology 2 domains of the regulatory subunit. We did not obtain evidence to support previous observations that the regulatory subunit of phosphatidylinositol 3 kinase is regulated through association with the tyrosine kinase Syk. The present results provide a molecular basis for the regulation of the p85/110 form of phosphatidylinositol 3-kinase by GPVI, the collagen receptor that underlies activation. PMID- 9852113 TI - The role of ubiquitin conjugation in glucose-induced proteolysis of Saccharomyces maltose permease. AB - In Saccharomyces, the addition of glucose induces a rapid degradation of maltose permease that is dependent on endocytosis and vacuolar proteolysis (Medintz, I., Jiang, H., Han, E. K., Cui, W., and Michels, C. A. (1996) J. Bacteriol. 178, 2245 2254). Here we report on the role of ubiquitin conjugation in this process. Deletion of DOA4, which causes decreased levels of available ubiquitin, severely decreases the rate of glucose-induced proteolysis, and this is suppressed by the overproduction of ubiquitin. Overexpression of ubiquitin in an endocytosis deficient end3-ts strain results in the glucose-stimulated accumulation of a larger molecular weight species of maltose permease, which we demonstrate is a ubiquitin-modified form of the protein by utilizing two ubiquitin alleles with different molecular weights. The size of this ubiquitinated species of maltose permease is consistent with monoubiquitination. A promoter mutation that reduces expression of RSP5/NPI1, a postulated ubiquitin-protein ligase, dramatically reduces the rate of glucose-induced proteolysis of maltose permease. The role of various ubiquitin-conjugating enzymes was investigated using strains carrying mutant alleles ubc1Delta ubc4Delta, ubc4Delta ubc5Delta, cdc34-ts2/ubc3, and ubc9 ts. Loss of these functions was not shown to effect glucose-induced proteolysis of maltose permease, but loss of Ubc1, -4, and -5 was found to inhibit maltose permease expression at the post-transcriptional level. PMID- 9852114 TI - Two adjacent protein binding motifs in the cbh2 (cellobiohydrolase II-encoding) promoter of the fungus Hypocrea jecorina (Trichoderma reesei) cooperate in the induction by cellulose. AB - The cellulase system of the filamentous fungus Hypocrea jecorina (Trichoderma reesei) consists of several cellobiohydrolases, endoglucanases, and beta glucosidases, encoded by separate genes, which are coordinately expressed in the presence of cellulose or the disaccharide sophorose. Using cell-free extracts from sophorose-induced and noninduced mycelia and various fragments of the cbh2 promoter of H. jecorina in electrophoretic mobility shift assay (EMSA) analysis and performing in vitro and in vivo footprinting analysis, we detected the nucleotide sequence 5'-ATTGGGTAATA-3' (consequently named cbh2-activating element (CAE)) to bind a protein complex with different migration in EMSA of induced and noninduced cell-free extracts. EMSA analysis, employing oligonucleotide fragments containing specifically mutated versions of CAE, revealed that protein binding requires the presence of an intact copy of either one of two adjacent motifs: a CCAAT (=ATTGG) box on the template strand and a GTAATA box on the coding strand, whereas a simultaneous mutation in both completely abolished binding. H. jecorina transformants, containing correspondingly mutated versions of the cbh2 promoter fused to the Escherichia coli hph gene as a reporter, expressed hph in a manner paralleling the efficacy of CAE-protein complex formation in EMSA, suggesting that the presence of either of both motifs is required for induction of cbh2 gene transcription. Antibody supershift experiments with anti-HapC antiserum as well as EMSA competition experiments with CCAAT binding promoter fragments of the Aspergillus nidulans amdS promoter suggest that the H. jecorina CCAAT box binding complex contains a homologue of HapC. The nature of the adjacent, GTAATA-binding protein(s) and its cooperation with the HapC homologue in cbh2 gene induction is discussed. PMID- 9852115 TI - Purification and characterization of a membrane bound neutral pH optimum magnesium-dependent and phosphatidylserine-stimulated sphingomyelinase from rat brain. AB - Sphingomyelin hydrolysis and ceramide generation catalyzed by sphingomyelinases (SMase) are key components of the signaling pathways in cytokine- and stress induced cellular responses. In this study, we report the partial purification and characterization of the membrane bound, neutral pH optimal, and magnesium dependent SMase (N-SMase) from rat brain. Proteins from Triton X-100 extract of brain membrane were purified sequentially with DEAE-Sephacel, heparin-Sepharose, ceramic hydroxyapatite, Mono Q, phenyl-Superose, and Superose 12 column chromatography. After eight purification steps, the specific activity of the enzyme increased by 3030-fold over the brain homogenate. The enzyme hydrolyzed sphingomyelin but not phosphatidylcholine and its activity was dependent upon magnesium with an optimal pH of 7.5 and a native pI of 5.2. Delipidation of the enzyme through chromatographic purification or by extraction with 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonic acid followed by gel filtration revealed that the enzyme became increasingly dependent on phosphatidylserine (PS). Up to 20-fold stimulation was observed with PS whereas other lipids examined were either ineffective or only mildly stimulatory. The Km of the enzyme for substrate sphingomyelin (3.4 mol %) was not affected by PS. The highly purified enzyme was inhibited by glutathione with a >95% inhibition observed with 3 mM glutathione and with a Hill number calculated at approximately 8. The significance of these results to the regulation of N-SMase is discussed. PMID- 9852116 TI - Role of cysteine residues in human angiotensinogen. Cys232 is required for angiotensinogen-pro major basic protein complex formation. AB - The M235T polymorphism of human angiotensinogen is associated with essential and pregnancy-induced hypertension. A covalent complex is formed between angiotensinogen and the proform of the eosinophil major basic protein (proMBP) during pregnancy. The sequence of human angiotensinogen contains four cysteines. Their function was analyzed. Presence of free cysteines was demonstrated by their alkylation with iodo[14C]acetic acid. A disulfide bond between Cys18 and Cys138 using a fully N-deglycosylated mutant of human angiotensinogen was identified by tryptic digestion and mass spectrometry. We produced angiotensinogen. proMBP complex by co-transfection of COS-7 cells and by co-culturing transfected CHO-K1 cells. Experiments with 8 mutated recombinant angiotensinogen, in which one or more of the four cysteines were replaced by alanine, demonstrated that Cys232 is involved in complex formation and could interact with the M235T variant. The angiotensinogen.proMBP complex was isolated by molecular sieving. Hydrolysis of the complex by human renin was 7 times slower than hydrolysis of monomeric form, whatever the M235T genotype. The complex:monomeric angiotensinogen ratio was greater for Met235 (72%) than for Thr235 (58%) angiotensinogen. These data suggest a new pathophysiological explanation for the genetic association between M235T angiotensinogen polymorphism and pregnancy-induced hypertension. PMID- 9852117 TI - Identification of serine 356 and serine 363 as the amino acids involved in etorphine-induced down-regulation of the mu-opioid receptor. AB - Agonist-induced internalization of G protein-coupled receptors is influenced by many structural determinants including the carboxyl tail. To investigate the role of serine and threonine residues within the carboxyl tail, several mutants were constructed by truncating the carboxyl tail of the hemagglutinin-tagged mu-opioid receptor, thereby removing serines and threonines systematically. Neuro2A cells stably expressing the truncated receptors did not exhibit a significant alteration in the affinity of [3H]diprenorphine or etorphine for the receptor or the potency of etorphine to inhibit forskolin-stimulated adenylyl cyclase activity. Chronic etorphine treatment resulted in a time-dependent down regulation of all the truncated receptors, except MOR1TAG355D, thus revealing the importance of the four amino acids between Ser355 and Glu359 (STIE). Surprisingly, deletion of the STIE sequence resulted in a receptor that down regulated the same as the wild-type receptor. The involvement of multiple amino acids within the carboxyl tail was demonstrated by combining alanine substitutions of several putative G-protein-coupled receptor kinase phosphorylation sites. Systematic analysis of these receptors indicated that mutation of Ser356 and Ser363 to alanine attenuated agonist-mediated down regulation. The magnitude of etorphine-induced phosphorylation of this mutant receptor, however, was similar to that of the wild-type mu-opioid receptor. Thus, phosphorylation of the carboxyl tail of the mu-opioid receptor is not an obligatory event for etorphine-induced down-regulation of the receptor. PMID- 9852118 TI - Attenuation of mammalian target of rapamycin activity by increased cAMP in 3T3-L1 adipocytes. AB - Incubating 3T3-L1 adipocytes with forskolin, which increases intracellular cAMP by activating adenylate cyclase, mimicked rapamycin by attenuating the effect of insulin on stimulating the phosphorylation of four (S/T)P sites in PHAS-I, a downstream target of the mammalian target of rapamycin (mTOR) signaling pathway. To investigate the hypothesis that increasing cAMP inhibits mTOR, the protein kinase activity of mTOR was measured in an immune complex assay with recombinant PHAS-I as substrate. Both forskolin and 8-(4-chlorophenylthio)adenosine 3'-5' monophosphate (CPT-cAMP) prevented the activation of mTOR by insulin in adipocytes, but neither agent affected mTOR activity when added directly to the immunopurified protein. In contrast, the cAMP phosphodiesterase inhibitor, theophylline, inhibited mTOR activity not only when added to intact adipocytes but also when added to immunopurified mTOR in vitro, demonstrating that certain methylxanthines are able to inhibit mTOR independently of increasing cAMP. Forskolin and CPT-cAMP blocked the effect of insulin on increasing mTOR phosphorylation, which was assessed using mTAb1, an antibody whose binding is inhibited by phosphorylation of mTOR. Although the mTAb1 epitope contains a consensus site for protein kinase B, neither agent inhibited the activation of protein kinase B produced by insulin. These findings support the interpretation that increasing cAMP attenuates the effects of insulin on PHAS-I, p70(S6K), and other downstream targets of the mTOR signaling pathway by inhibiting the phosphorylation and activation of mTOR. PMID- 9852119 TI - Rabbit lung indolethylamine N-methyltransferase. cDNA and gene cloning and characterization. AB - Indolethylamine N-methyltransferase (INMT) catalyzes the N-methylation of tryptamine and structurally related compounds. This reaction has been studied because of its possible role in the in vivo synthesis of psychoactive compounds or neurotoxins and has been characterized biochemically in preparations of rabbit lung. Therefore, we set out to purify rabbit lung INMT, to clone and express its cDNA, and to clone and structurally characterize its gene as steps toward understanding the function and regulation of this enzyme. Rabbit lung INMT was purified and partial amino acid sequence was obtained. A polymerase chain reaction-based approach was then used to clone a rabbit lung INMT cDNA with a 792 base pair open reading frame that encoded a 263-amino acid protein with a predicted molecular mass of 29 kDa. When the cDNA was expressed in COS-1 cells, the encoded protein catalyzed the methylation of tryptamine and structurally related compounds, and was inhibited by two products of the reaction, S-adenosyl L-homocysteine (AdoHcy) and N,N-dimethyltryptamine, as well as antimigraine drugs that are structurally related to N,N-dimethyltryptamine. Northern blot analysis demonstrated the presence of 2.0-kilobase mRNA species in rabbit lung, liver and, at lower levels, in brain. The cDNA was then used to clone the rabbit INMT gene. That gene had three exons and was structurally similar to the genes for nicotinamide N-methyltransferase and phenylethanolamine N-methyltransferase in several species. Cloning and expression of a rabbit lung INMT cDNA and cloning of the rabbit INMT gene represent important steps toward determination of the function and regulation of this mammalian methyltransferase enzyme. PMID- 9852120 TI - Template activating factor-I remodels the chromatin structure and stimulates transcription from the chromatin template. AB - To study the mechanisms of replication and transcription on chromatin, we have been using the adenovirus DNA complexed with viral basic core proteins, called Ad core. We have identified template activating factor (TAF)-I from uninfected HeLa cells as the factor that stimulates replication and transcription from the Ad core. The nuclease sensitivity assays have revealed that TAF-I remodels the Ad core, thereby making transcription and replication apparatus accessible to the template DNA. To examine whether TAF-I remodels the chromatin consisting of histones, the chromatin structure was reconstituted on the DNA fragment with core histones by the salt dialysis method. The transcription from the reconstituted chromatin was completely repressed, while TAF-I remodeled the chromatin and stimulated the transcription. TAF-I was found to interact with histones. Furthermore, it was shown that TAF-I is capable not only of disrupting the chromatin structure but also of preventing the formation of DNA-histone aggregation and transferring histones to naked DNA. The possible function of TAF I in conjunction with a histone chaperone activity is discussed. PMID- 9852121 TI - Differential regulation of phospholipase A2 (PLA2)-dependent Ca2+ signaling in smooth muscle by cAMP- and cGMP-dependent protein kinases. Inhibitory phosphorylation of PLA2 by cyclic nucleotide-dependent protein kinases. AB - Both cAMP- and cGMP-dependent protein kinases inhibit agonist-stimulated phospholipase C-beta (PLC-beta) activity and inositol 1,4,5-trisphosphate dependent Ca2+ release in vascular and visceral smooth muscle. In smooth muscle of the intestinal longitudinal layer, however, the initial steps in Ca2+ mobilization involve activation of cytosolic PLA2 (cPLA2) and arachidonic acid (AA)-dependent stimulation of Ca2+ influx. The present study examined whether cAMP- and cGMP-dependent protein kinases are capable of regulating these processes also. Agents that activated cAMP-dependent protein kinase (5, 6 dichloro-1-beta-D-ribofuranosylbenzimidazole 3',5'-cyclic monophosphothioate (Sp isomer) and isoproterenol), cGMP-dependent protein kinase (8-(4-chlorophenylthio) guanosine 3',5'-cyclic monophosphate and Na nitroprusside), or both kinases (vasoactive intestinal peptide and isoproterenol >1 microM) induced phosphorylation of cPLA2 and inhibition of agonist-stimulated cPLA2 activity. Phosphorylation and inhibition of cPLA2 activity by cAMP- and cGMP-dependent protein kinases were blocked by the corresponding selective inhibitors (cAMP dependent protein kinase, N-[2(p-bromocinnamylamino)ethyl]-5-isoquinoline sulfonamide hydrochloride (H-89) and myristoylated protein kinase inhibitor () amide; cGMP-dependent protein kinase, (8R,9S, 11S)-(-)-9-methoxy-carbamyl-8 methyl-2,3,9,10-tetrahydro-8, 11-epoxy-1H,8H,11H,-2,7b,11a trizadizobenzo(a,g)cycloocta(c, d, e)-trinden-1-one (KT-5823)). In contrast, AA stimulated Ca2+ influx was inhibited by agents that activated cGMP-dependent protein kinase only; the inhibition was selectively blocked by KT-5823. The study provides the first evidence of inhibitory phosphorylation of cPLA2 in vivo by cAMP- and cGMP-dependent protein kinases. Inhibition of cPLA2 activity and AA induced Ca2+ influx partly account for the ability of cAMP-dependent protein kinase and/or cGMP-dependent protein kinase to cause relaxation. Their importance resides in their location at the inception of the Ca2+ signaling cascade. PMID- 9852122 TI - Identification of the half-cystine residues in porcine submaxillary mucin critical for multimerization through the D-domains. Roles of the CGLCG motif in the D1- and D3-domains. AB - Plasmids encoding the amino-terminal region of porcine submaxillary mucin were modified by site-specific mutagenesis to assess the roles of individual half cystine residues in the assembly of disulfide-linked multimers of mucin. COS-7 cells with the plasmid containing C1199A expressed primarily monomers, suggesting that half-cystine 1199 in the D3-domain is involved in forming mucin multimers. This residue is in the sequence C1199SWRYEPCG, which is highly conserved in the D3-domain of other secreted mucins and human prepro-von Willebrand factor. In contrast, cells with the plasmid containing C1276A expressed trimers like those with unmutated plasmid, suggesting that half-cystine 1276 is not involved in formation of disulfide-bonded multimers. The roles of the half-cystines in the CGLCG motifs in the assembly of disulfide-bonded multimers of mucin were also assessed. Cells with plasmids in which both half-cystines in the motif in the D1- or D3-domain of mucin are replaced by alanine expressed proteins that were poorly secreted, suggesting that these mutations impair normal folding of the expressed proteins. A plasmid with a mutant D1-domain motif expressed monomers, whereas one with a mutant D3-domain motif expressed monomers and trimers. However, the trimers expressed by the latter plasmid were assembled in non-acidic compartments, as judged by expression studies in the presence of monensin, which inhibits trimer formation by unmutated plasmid, but not by the mutant plasmid. These results suggest that the CGLCG motif in the D1-domain is required for multimerization in the trans-Golgi complex. However, the CGLCG motif in the D3 domain appears to prevent formation of mucin multimers in non-acidic compartments of the cell. Plasmids encoding the D1- and D2-domains, the D1- and D3-domains, or only the D3-domain also expressed oligomers in the presence of monensin, suggesting that the three D-domains must be contiguous to avoid multimerization in non-acidic compartments. It is possible that these motifs in mucins are engaged in the thiol-disulfide interchange reactions during the assembly of disulfide-bonded multimers of mucin. PMID- 9852123 TI - Negative control of the poly(A)-binding protein mRNA translation is mediated by the adenine-rich region of its 5'-untranslated region. AB - Translation of the mRNA for the poly(A)-binding protein (PABP) may be autoregulated by the binding of PABP to the A-rich segment of its 5'-untranslated region (UTR). To test this hypothesis, we examined the effect of different fragments of the 5'-UTR from human PABP cDNA on the translation of the beta galactosidase (beta-Gal) reporter gene. Presence of the A-rich sequence from the 5'-UTR of PABP mRNA inhibited expression of the chimeric beta-Gal gene in transfected HeLa cells. The differences in expression of beta-Gal polypeptide was due to the translational repression of beta-Gal mRNA containing the A-rich 5'-UTR of PABP mRNA. The A-rich region of the 5'-UTR located within nucleotides 58-146 of PABP mRNA was sufficient to mediate translational control of this mRNA expression. We also examined the effect of overexpression of PABP mRNA in HeLa cells. The ectopic PABP mRNA without the A-rich 5'-UTR region was translated efficiently, whereas the translation of the endogenous PABP mRNA was substantially inhibited in the transfected cells. In contrast, the ectopic PABP mRNA containing the A-rich 5'-UTR region did not show similar effect on the translation of the endogenous PABP mRNA in these cells. These results suggest that feedback control of mRNA translation is involved in regulating PABP expression in HeLa cells. PMID- 9852124 TI - Insulin receptor substrate 2 and Shc play different roles in insulin-like growth factor I signaling. AB - The major substrates for the type I insulin-like growth factor (IGF-I) receptor are Shc and insulin receptor substrate (IRS) proteins. In the current study, we report that IGF-I induces a sustained tyrosine phosphorylation of Shc and its association with Grb2 in SH-SY5Y human neuroblastoma cells. The time course of Shc tyrosine phosphorylation parallels the time course of IGF-I-stimulated activation of extracellular signal-regulated kinase (ERK). Transfection of SH SY5Y cells with a p52 Shc mutant decreases Shc tyrosine phosphorylation and Shc Grb2 association. This results in the inhibition of IGF-I-mediated ERK tyrosine phosphorylation and neurite outgrowth. In contrast, IGF-I induces a transient tyrosine phosphorylation of IRS-2 and an association of IRS-2 with Grb2. The time course of IRS-2 tyrosine phosphorylation and IRS-2-Grb2 and IRS-2-p85 association closely resembles the time course of IGF-I-mediated membrane ruffling. Treating cells with the phosphatidylinositol 3'-kinase inhibitors wortmannin and LY294002 blocks IGF-I-induced membrane ruffling. The ERK kinase inhibitor PD98059, as well as transfection with the p52 Shc mutant, has no effect on IGF-I-mediated membrane ruffling. Immunolocalization studies show IRS-2 and Grb2, but not Shc, concentrated at the tip of the extending growth cone where membrane ruffling is most active. Collectively, these results suggest that the association of Shc with Grb2 is essential for IGF-I-mediated neurite outgrowth, whereas the IRS-2-Grb2 phosphatidylinositol 3'-kinase complex may regulate growth cone extension and membrane ruffling. PMID- 9852125 TI - Overlapping cis sites used for splicing of HIV-1 env/nef and rev mRNAs. AB - Alternative splicing is used to generate more than 30 human immunodeficiency virus type 1 (HIV-1) spliced and unspliced mRNAs from a single primary transcript. The abundance of HIV-1 mRNAs is determined by the efficiencies with which its different 5' and 3' splice sites are used. Three splice sites (A4c, A4a, and A4b) are upstream of the rev initiator AUG. RNAs spliced at A4c, A4a, and A4b are used as mRNAs for Rev. Another 3' splice site (A5) is immediately downstream of the rev initiator. RNAs spliced at A5 are used as mRNAs for Env and Nef. In this report, primer extension analysis of splicing intermediates was used to show that there are eight branch points in this region, all of which map to adenosine residues. In addition, cis elements recognized by the cellular splicing machinery overlap; the two most 3' branch points overlap with the AG dinucleotides at rev 3' splice sites A4a and A4b. Competition of the overlapping cis sites for different splicing factors may play a role in maintaining the appropriate balance of mRNAs in HIV-1-infected cells. In support of this possibility, mutations at rev 3' splice site A4b AG dinucleotide dramatically increased splicing of the env/nef 3' splice site A5. This correlated with increased usage of the four most 3' branch points, which include those within the rev 3' splice site AG dinucleotides. Consistent with these results, analysis of a mutant in which three of the four env/nef branch points were inactivated indicated that use of splice site A5 was inhibited and splicing was shifted predominantly to the most 5' rev 3' splice site A4c with preferential use of the two most 5' branch points. Our results suggest that spliceosomes formed at rev A4a-4b, rev A4c, and env/nef A5 3' splice sites each recognize different subsets of the eight branch point sequences. PMID- 9852126 TI - Identification of the sites of N-linked glycosylation on the human calcium receptor and assessment of their role in cell surface expression and signal transduction. AB - The human calcium receptor (hCaR) is a G-protein-coupled receptor containing 11 potential N-linked glycosylation sites in the large extracellular domain. The number of potential N-linked glycosylation sites actually modified, and the effect on cell surface expression and signal transduction of blocking glycosylation at these sites, was examined by site-directed mutagenesis. Asparagine residues of the consensus sequences (Asn-Xaa-Ser/Thr) for N-linked glycosylation were mutated to glutamine individually and in various combinations to disrupt the potential N-linked glycosylation sites in the context of the full length receptor. The cDNA constructs were transiently transfected into HEK-293 cells lacking endogeneous hCaR, and expressed receptors were analyzed by mobility differences on immunoblots, glycosidase digestion, intact cell enzyme-linked immunoassay, and extracellular calcium-stimulated phosphoinositide hydrolysis assay. Immunoblot analyses and glycosidase digestion studies of the wild type versus mutant receptors demonstrate that, of the 11 potential sites for N-linked glycosylation, eight sites (Asn-90, -130, -261, -287, -446, -468, -488, and -541) are glycosylated; the three remaining sites (Asn-386, -400, and -594) may not be efficiently glycosylated in the native receptor. Sequential mutagenesis of multiple N-linked glycosylation sites and analyses by immunoblotting, immunofluorescence, biotinylation of cell surface proteins, and intact cell enzyme-linked immunoassay indicated that disruption of as few as three glycosylation sites impairs proper processing and expression of the receptor at the cell surface. Disruption of five glycosylation sites reduced cell surface expression by 50-90% depending on which five sites were disrupted. Phosphoinositide hydrolysis assay results for various glycosylation-defective mutant receptors in general correlated well with the level of cell surface expression. Our results demonstrate that among 11 potential N-linked glycosylation sites on the hCaR, eight sites are actually utilized; glycosylation of at least three sites is critical for cell surface expression of the receptor, but glycosylation does not appear to be critical for signal transduction. PMID- 9852127 TI - Purification of the lysosomal sialic acid transporter. Functional characteristics of a monocarboxylate transporter. AB - Sialic acid and glucuronic acid are monocarboxylated monosaccharides, which are normally present in sugar side chains of glycoproteins, glycolipids, and glycosaminoglycans. After degradation of these compounds in lysosomes, the free monosaccharides are released from the lysosome by a specific membrane transport system. This transport system is deficient in the human hereditary lysosomal sialic acid storage diseases (Salla disease and infantile sialic acid storage disease, OMIM 269920). The lysosomal sialic acid transporter from rat liver has now been purified to apparent homogeneity in a reconstitutively active form by a combination of hydroxyapatite, lectin, and ion exchange chromatography. A 57-kDa protein correlated with transport activity. The transporter recognized structurally different types of acidic monosaccharides, like sialic acid, glucuronic acid, and iduronic acid. Transport of glucuronic acid was inhibited by a number of aliphatic monocarboxylates (i.e. lactate, pyruvate, and valproate), substituted monocarboxylates, and several dicarboxylates. cis-Inhibition, trans stimulation, and competitive inhibition experiments with radiolabeled glucuronic acid as well as radiolabeled L-lactate demonstrated that L-lactate is transported by the lysosomal sialic acid transporter. L-Lactate transport was proton gradient dependent, saturable with a Km of 0.4 mM, and mediated by a single mechanism. These data show striking biochemical and structural similarities of the lysosomal sialic acid transporter with the known monocarboxylate transporters of the plasma membrane (MCT1, MCT2, MCT3, and Mev). PMID- 9852128 TI - pH-dependent gating of ROMK (Kir1.1) channels involves conformational changes in both N and C termini. AB - ROMK channels (Kir1.1) are members of the superfamily of inward rectifier potassium channels (Kir) and represent the channels underlying K+ secretion in the kidney. As their native counterparts, Kir1.1 channels are gated by intracellular pH, with acidification leading to channel closure. Although a lysine residue (Lys80) close to the first hydrophobic segment M1 has been identified as the pH sensor, little is known about how opening and closing of the channel is accomplished. Here we investigate the gating process of Kir1.1 channels exploiting their state-dependent modification by water-soluble oxidants and sulfhydryl reagents. Mutagenesis of all intracellular cysteines either alone or in combination revealed two residues targeted by these reagents, one in the N terminus (Cys49) and one in the C terminus (Cys308) of the channel protein. Both sites reacted with the thiol reagents only in the closed state and not in the open state. These results indicate that pH-dependent gating of Kir1.1 channels involves movement of protein domains in both N and C termini of the Kir1.1 protein. PMID- 9852129 TI - Localization of the Rab3 small G protein regulators in nerve terminals and their involvement in Ca2+-dependent exocytosis. AB - The Rab3 small G protein subfamily (Rab3) consists of four members, Rab3A, -B, C, and -D. We have recently isolated and characterized the Rab3 regulators, GDP/GTP exchange protein (GEP) and GTPase activating protein (GAP), both of which are specific for the Rab3 subfamily. Rab3 GEP stimulates the conversion of the GDP-bound inactive form to the GTP-bound active form, whereas Rab3 GAP stimulates the reverse reaction. Of the four members of the Rab3 subfamily, evidence is accumulating that Rab3A is involved in Ca2+-dependent exocytosis, particularly in neurotransmitter release. We first analyzed the subcellular localization of Rab3 GEP and GAP in rat brain. Subcellular fractionation analysis showed that both Rab3 GEP and GAP were enriched in the synaptic soluble fraction. Immunocytochemical analysis in primary cultured rat hippocampal neurons showed that both Rab3 GEP and GAP were concentrated at the presynaptic nerve terminals. We then examined whether Rab3 GEP and GAP were involved in Ca2+-dependent exocytosis by use of human growth hormone (GH) co-expression assay system of cultured PC12 cells. Overexpression of the deletion mutant of Rab3 GEP possessing the catalytic activity reduced the high K+-induced GH release without affecting the basal GH release, whereas that of the deletion mutant lacking the catalytic activity showed no effect on the high K+-induced GH release. In contrast, overexpression of Rab3 GAP or its deletion mutant possessing the catalytic activity did not affect the high K+-induced GH release or the basal GH release. These results indicate that Rab3 GEP and GAP are colocalized with Rab3A at the synaptic release sites and suggest that they regulate the activity of Rab3A and are involved in Ca2+-dependent exocytosis. PMID- 9852130 TI - In vivo autopolysialylation and localization of the polysialyltransferases PST and STX. AB - A select group of mammalian proteins have been shown to possess alpha2,8 polysialylated oligosaccharide chains. The best studied of these proteins is the neural cell adhesion molecule (NCAM). Polysialylation of NCAM has been shown to decrease NCAM-dependent and independent cell adhesion. PST (ST8Sia IV) and STX (ST8Sia II) are the two polysialyltransferases responsible for NCAM polysialylation. Recent studies revealed that PST itself is autopolysialylated in vitro (Muhlenhoff, M., Eckhardt, M., Bethe, A., Frosch, M., and Gerardy-Schahn, R. (1996) EMBO J. 15, 6943-6950). Here we report studies on the biosynthesis and localization of the PST and STX polysialyltransferases. Both PST and STX are expressed as high molecular mass, polydisperse forms that are associated with the cell and found soluble in the medium. Analysis of these high molecular mass forms by glycosidase digestion and serial immunoprecipitation/immunoblot experiments demonstrated that PST and STX are autopolysialylated in vivo. Indirect immunofluorescence microscopy and immunoprecipitation analyses demonstrated that autopolysialylated PST and STX are localized in the Golgi, on the cell surface, and in the extracellular space. The cell surface and extracellular localization of these polysialylated polysialyltransferases suggest that their polysialic acid chains, like those of NCAM, may modulate cell interactions. PMID- 9852132 TI - Multiple forms of the U2 small nuclear ribonucleoprotein auxiliary factor U2AF subunits expressed in higher plants. AB - Requirements for intron recognition during pre-mRNA splicing in plants differ from those in vertebrates and yeast. Plant introns contain neither conserved branch points nor distinct 3' splice site-proximal polypyrimidine tracts characteristic of the yeast and vertebrate introns, respectively. However, they are strongly enriched in U residues throughout the intron, property essential for splicing. To understand the roles of different sequence elements in splicing, we are characterizing proteins involved in intron recognition in plants. In this work we show that Nicotiana plumbaginifolia, a dicotyledonous plant, contains two genes encoding different homologs of the large 50-65-kDa subunit of the polypyrimidine tract binding factor U2AF, characterized previously in animals and Schizosaccharomyces pombe. Both plant U2AF65 isoforms, referred to as NpU2AF65a and NpU2AF65b, support splicing of an adenovirus pre-mRNA in HeLa cell nuclear extracts depleted of the endogenous U2AF factor. Both proteins interact with RNA fragments containing plant introns and show affinity for poly(U) and, to a lesser extend, poly(C) and poly(G). The branch point or the 3' splice site regions do not contribute significantly to intron recognition by NpU2AF65. The existence of multiple isoforms of U2AF may be quite general in plants because two genes expressing U2AF65 have been identified in Arabidopsis, and different isoforms of the U2AF small subunit are expressed in rice. PMID- 9852131 TI - Mitosis-specific phosphorylation and subcellular redistribution of the RIIalpha regulatory subunit of cAMP-dependent protein kinase. AB - Phosphorylation of the RII regulatory subunits of cyclic AMP-dependent protein kinases (PKAs) was examined during the HeLa cell cycle. Three RIIalpha isoforms of 51, 54, and 57 kDa were identified by RIIalpha immunodetection and labeling with 8-azido[32P]cAMP in different cell cycle phases. These isoforms were characterized as different phosphorylation states by the use of selective PKA and cyclin-directed kinase inhibitors. Whereas RIIalpha autophosphorylation by PKA caused RIIalpha to shift from 51 to 54 kDa, phosphorylation of RIIalpha by one other or a combination of several kinases activated during mitosis caused RIIalpha to shift from 51 to 57 kDa. In vivo incorporation of [32P]orthophosphate into mitotic cells and RIIalpha immunoprecipitation demonstrated that RIIalpha was hyperphosphorylated on a different site than the one phosphorylated by PKA. Deletion and mutation analysis demonstrated that the cyclin B-p34(cdc2) kinase (CDK1) phosphorylated human recombinant RIIalpha in vitro on Thr54. Whereas RIIalpha was associated with the Golgi-centrosomal region during interphase, it was dissociated from its centrosomal localization at metaphase-anaphase transition. Furthermore, particulate RIIalpha from HeLa cell extracts was solubilized following incubation with CDK1 in vitro. Our results suggest that at the onset of mitosis, CDK1 phosphorylates RIIalpha, and this may alter its subcellular localization. PMID- 9852134 TI - Characterization of dominant negative arrestins that inhibit beta2-adrenergic receptor internalization by distinct mechanisms. AB - Arrestins have been shown to act as adaptor proteins that mediate the interaction of G protein-coupled receptors with the endocytic machinery. In this study, the role of arrestin-3 in receptor internalization was investigated by constructing different arrestin-3 minigenes that could potentially act as dominant negative inhibitors of arrestin function. Expression of arrestin-3 proteins containing amino acids 1-320 or 201-409 resulted in the inhibition of beta2-adrenergic receptor internalization in HEK-293 cells by approximately 40%. Both of these arrestins were diffusely localized within the cytoplasm of transfected cells, were unable to mediate redistribution of receptors to clathrin-coated pits, and did not localize to coated pits in either the presence or absence of receptor and agonist. Arrestin-3(1-320), but not arrestin-3(201-409), bound to light-activated phosphorylated rhodopsin with an affinity comparable with that of wild-type arrestin-3. In contrast, expression of arrestin-3 proteins composed of only the clathrin binding domain, arrestin-3(284-409), and arrestin-3(290-409) resulted in the constitutive localization of these arrestins to coated pits. Arrestin-3(284 409) and arrestin-3(290-409) acted as dominant negative inhibitors of wild-type arrestin function, inhibiting receptor internalization by 70 and 30%, respectively. Carboxyl-terminal deletions of arrestin-3 retained the ability to promote internalization until residues amino-terminal to amino acid 350 were deleted, suggesting that residues in this region also compose part of the clathrin binding domain in addition to the major binding site between residues 371-379. These studies characterize at least two distinct mechanisms, competition for either receptor or clathrin binding, by which dominant negative arrestins inhibit receptor internalization and further define residues within arrestin-3 that constitute the clathrin binding domain. PMID- 9852133 TI - BMCP1, a novel mitochondrial carrier with high expression in the central nervous system of humans and rodents, and respiration uncoupling activity in recombinant yeast. AB - We report here the cloning and functional analysis of a novel homologue of the mitochondrial carriers predominantly expressed in the central nervous system and referred to as BMCP1 (brain mitochondrial carrier protein-1). The predicted amino acid sequence of this novel mitochondrial carrier indicates a level of identity of 39, 31, or 30%, toward the mitochondrial oxoglutarate carrier, phosphate carrier, or adenine nucleotide translocator, respectively, and a level of identity of 34, 38, or 39% with the mitochondrial uncoupling proteins UCP1, UCP2, or UCP3, respectively. Northern analysis of mouse, rat, or human tissues demonstrated that mRNA of this novel gene is mainly expressed in brain, although it is 10-30-fold less expressed in other tissues. In situ hybridization analysis of brain showed it is particularly abundant in cortex, hippocampus, thalamus, amygdala, and hypothalamus. Chromosomal mapping indicates that BMCP1 is located on chromosome X of mice and at Xq24 in man. Expression of the protein in yeast strongly impaired growth rate. Analysis of respiration of total recombinant yeast or yeast spheroplasts and in particular of the relationship between respiratory rate and membrane potential of yeast spheroplasts revealed a marked uncoupling activity of respiration, suggesting that although BMCP1 sequence is more distant from the uncoupling proteins (UCPs), this protein could be a fourth member of the UCP family. PMID- 9852135 TI - The identification of a cis-element and a trans-acting factor involved in the response to polyamines and polyamine analogues in the regulation of the human spermidine/spermine N1-acetyltransferase gene transcription. AB - The superinduction of spermidine/spermine N1-acetyltransferase (SSAT) gene has been associated with a cytotoxic response to a new class of antineoplastic polyamine analogues. The initial mechanism of SSAT superinduction is an increase in transcription in response to analogue exposure. This increased transcription appears to be modulated through the association between a nuclear protein factor and a cis-element described here as the polyamine-responsive element (PRE). The PRE was identified as a 9-base pair sequence, 5'-TATGACTAA-3', in the context of a 31-base pair stretch from -1522 to -1492 base pairs with respect to the SSAT transcriptional start site. This element binds a nuclear factor from polyamine analogue-responsive cells, but not from polyamine analogue-insensitive cells. The labeled PRE was used to clone and identify the transcription factor, Nrf-2, that binds constitutively to the PRE sequence. Although the PRE sequence shares homology to the originally identified Nrf-2 recognition sequence, the two sequences are not identical. The Nrf-2 transcription factor appears only to be present in cell types that are capable of expressing high amounts of SSAT. The results of these studies suggest that Nrf-2, bound to the PRE, plays an important regulatory role of expression of the human SSAT gene. PMID- 9852136 TI - The function of the p190 Rho GTPase-activating protein is controlled by its N terminal GTP binding domain. AB - p190 is a GTPase-activating protein (GAP) for the Rho family of GTPases. The GAP domain of p190 is at the C terminus of the protein. At its N terminus, p190 contains a GTP binding domain of unknown significance. We have introduced a mutation (Ser36 --> Asn) into this domain of p190 that decreased its ability to bind guanine nucleotide when expressed as a hemagglutinin (HA)-tagged protein in COS cells. In vitro, both the wild type and S36N mutant HA-p190 proteins showed similar GAP activities toward RhoA, but when expressed in NIH 3T3 fibroblasts only wild type p190 appeared able to function as a RhoGAP. Wild type HA-p190 induced a phenotype of rounded cells with long, beaded extensions similar to that seen when Rho function is disrupted by ADP-ribosylation. HA-p190(S36N), although expressed at a similar level to the wild type protein, had no discernible effect on the cells. The beaded extension phenotype induced by wild type HA-p190 required GAP function. A GAP-defective mutant, p190(R1283A), had no effect on cell morphology. Moreover, the beaded extension phenotype could be suppressed by co-expression of a gain-of-function Rho mutant, RhoA(G14V), or Rac mutant, Rac1(G12V). Activation of the Jun kinase (JNK) via muscarinic receptors was inhibited by wild type HA-p190, but JNK activity was enhanced by the S36N mutant. Co-expression of HA-p190 with a fragment containing only the mutated GTP binding domain partially inhibited the beaded extension phenotype, suggesting that it may sequester a factor required for p190 function. Taken together these data demonstrate that within the cell, the Rho/Rac GAP activity of p190 can be regulated by the N-terminal GTP binding domain. PMID- 9852138 TI - The coactivator CBP stimulates human T-cell lymphotrophic virus type I Tax transactivation in vitro. AB - Tax interacts with the cellular cyclic AMP-responsive element binding protein (CREB) and facilitates the binding of the coactivator CREB binding protein (CBP), forming a multimeric complex on the cyclic AMP-responsive element (CRE)-like sites in the human T-cell lymphotrophic virus type I (HTLV-I) promoter. The trimeric complex is believed to recruit additional regulatory proteins to the HTLV-I long terminal repeat, but there has been no direct evidence that CBP is required for Tax-mediated transactivation. We present evidence that Tax and CBP activate transcription from the HTLV-I 21 base pair repeats on naked DNA templates. Transcriptional activation of the HTLV-I sequences required both Tax and CBP and could be mediated by either the N-terminal activation domain of CBP or the full-length protein. Fluorescence polarization binding assays indicated that CBP does not markedly enhance the affinity of Tax for the trimeric complex. Transcription analyses suggest that CBP activates Tax-dependent transcription by promoting transcriptional initiation and reinitiation. The ability of CBP to activate the HTLV-I promoter does not involve the stabilization of Tax binding, but rather depends upon gene activation properties of the co-activator that function in the context of a naked DNA template. PMID- 9852137 TI - Protein kinase Cdelta-dependent induction of manganese superoxide dismutase gene expression by microtubule-active anticancer drugs. AB - Bacterial lipopolysaccharide can induce manganese superoxide dismutase (MnSOD) gene expression in a variety of cells. Paclitaxel (taxol) shares many properties of lipopolysaccharide. Here we report that paclitaxel can induce MnSOD gene expression in human lung adenocarcinoma cell line A549 in a time- and dose dependent manner. Additional anticancer drugs, vinblastine and vincristine, also induced MnSOD gene expression. We have shown previously (Das, K. C., and White, C. W. (1997) J. Biol. Chem. 272, 14914-14920) that these drugs can activate protein kinase C (PKC). The PKC agonists thymeleatoxin (0.5 microM) and 12 deoxyphorbol 13-phenylacetate 20-acetate (dPPA; 10 nM) potently induced MnSOD gene expression. Calphostin C and GF109203X, both specific inhibitors of PKC, each inhibited MnSOD gene expression by anticancer agents. Down-regulation of PKC by prolonged treatment with phorbol 12-myristate 13-acetate (PMA) also inhibited induction of MnSOD by anticancer drugs, indicating an important role of PKC in MnSOD signaling by these agents. Of 11 PKC isoenzymes, only PKCdelta translocated to the cell membrane after stimulation with anticancer drugs. By contrast, dPPA, PMA, and thymeleatoxin caused translocation of PKCalpha, betaI, delta, and mu isotypes. Anticancer drug-stimulated cells also had increased total PKC activity in membrane and cytosolic fractions. Thus, paclitaxel, vinblastine, and vincristine each specifically activate PKCdelta, whereas PMA, thymeleatoxin, and dPPA activate multiple isoenzymes. PKCdelta was the only isoform activated by each agent in both groups of compounds effective in MnSOD induction. PMID- 9852139 TI - Promoter structure and transcriptional activation with chromatin templates assembled in vitro. A single Gal4-VP16 dimer binds to chromatin or to DNA with comparable affinity. AB - To gain a better understanding of the role of chromatin in the regulation of transcription by RNA polymerase II, we examined the relation between promoter structure and the ability of Gal4-VP16 to function with chromatin templates assembled in vitro. First, to investigate whether there are synergistic interactions among multiple bound factors, we studied promoter constructions containing one or five Gal4 sites and found that a single recognition site is sufficient for Gal4-VP16 to bind to chromatin, to induce nucleosome rearrangement, and to activate transcription. Notably, we observed that Gal4-VP16 binds to a single site in chromatin with affinity comparable with that which it binds to naked DNA, even in the absence of ATP-dependent nucleosome remodeling activity. Second, to explore the relation between translational nucleosome positioning and transcriptional activation, we analyzed a series of promoter constructions in which nucleosomes were positioned by Gal4-VP16 at different locations relative to the RNA start site. These experiments revealed that the positioning of a nucleosome over the RNA start site is not an absolute barrier to transcriptional activation. Third, to determine the contribution of core promoter elements to transcriptional activation with chromatin templates, we tested the ability of Gal4-VP16 to activate transcription with TATA box- versus DPE-driven core promoters and found that the TATA box is not required to achieve transcriptional activation by Gal4-VP16 with chromatin templates. These results suggest that a single protomer of a strong activator is able to bind to chromatin, to induce nucleosome remodeling, and to activate transcription in conjunction with a broad range of chromatin structures and core promoter elements. PMID- 9852141 TI - Chromosomal proteins HMG-14 and HMG-17 are released from mitotic chromosomes and imported into the nucleus by active transport. AB - The high mobility group 14/17 (HMG-14/-17) proteins form specific complexes with nucleosome core particles and produce distinct footprints on nucleosomal DNA. Therefore, they could be an integral part of the chromatin fiber. Here we show that during the cell cycle these proteins are transiently dissociated from chromatin. They colocalize with the nuclear DNA in interphase and prophase but not in metaphase and anaphase. They relocate into the nucleus and colocalize again with the DNA in late telophase, concomitantly with the appearance of the nuclear envelope. Thus, these nucleosomal binding proteins are not always associated with chromatin. Using reconstituted nuclei and permeabilized cells, we demonstrate that these two small proteins, with a molecular mass <10 kD, are actively imported into the nucleus. We identify the major elements involved in the nuclear import of these chromosomal proteins: HMG-14/-17 proteins contain an intrinsic bipartite nuclear localization signal, and their entry into the nucleus through nuclear pores requires energy and the participation of importin alpha. These findings suggest that the cell cycle-related association of HMG-14/-17 with chromatin is dependent on, and perhaps regulated by, nuclear import processes. PMID- 9852140 TI - Spatial and temporal dynamics of DNA replication sites in mammalian cells. AB - Fluorescence microscopic analysis of newly replicated DNA has revealed discrete granular sites of replication (RS). The average size and number of replication sites from early to mid S-phase suggest that each RS contains numerous replicons clustered together. We are using fluorescence laser scanning confocal microscopy in conjunction with multidimensional image analysis to gain more precise information about RS and their spatial-temporal dynamics. Using a newly improved imaging segmentation program, we report an average of approximately 1,100 RS after a 5-min pulse labeling of 3T3 mouse fibroblast cells in early S-phase. Pulse-chase-pulse double labeling experiments reveal that RS take approximately 45 min to complete replication. Appropriate calculations suggest that each RS contains an average of 1 mbp of DNA or approximately 6 average-sized replicons. Double pulse-double chase experiments demonstrate that the DNA sequences replicated at individual RS are precisely maintained temporally and spatially as the cell progresses through the cell cycle and into subsequent generations. By labeling replicated DNA at the G1/S borders for two consecutive cell generations, we show that the DNA synthesized at early S-phase is replicated at the same time and sites in the next round of replication. PMID- 9852142 TI - pEg7, a new Xenopus protein required for mitotic chromosome condensation in egg extracts. AB - We have isolated a cDNA, Eg7, corresponding to a Xenopus maternal mRNA, which is polyadenylated in mature oocytes and deadenylated in early embryos. This maternal mRNA encodes a protein, pEg7, whose expression is strongly increased during oocyte maturation. The tissue and cell expression pattern of pEg7 indicates that this protein is only readily detected in cultured cells and germ cells. Immunolocalization in Xenopus cultured cells indicates that pEg7 concentrates onto chromosomes during mitosis. A similar localization of pEg7 is observed when sperm chromatin is allowed to form mitotic chromosomes in cytostatic factor arrested egg extracts. Incubating these extracts with antibodies directed against two distinct parts of pEg7 provokes a strong inhibition of the condensation and resolution of mitotic chromosomes. Biochemical experiments show that pEg7 associates with Xenopus chromosome-associated polypeptides C and E, two components of the 13S condensin. PMID- 9852143 TI - A novel nuclear import pathway for the transcription factor TFIIS. AB - We have identified a novel pathway for protein import into the nucleus. We have shown that the previously identified but uncharacterized yeast protein Nmd5p functions as a karyopherin. It was therefore designated Kap119p (karyopherin with Mr of 119 kD). We localized Kap119p to both the nucleus and the cytoplasm. We identified the transcription elongation factor TFIIS as its major cognate import substrate. The cytoplasmic Kap119p exists as an approximately stoichiometric complex with TFIIS. RanGTP, not RanGDP, dissociated the isolated Kap119p/TFIIS complex and bound to Kap119p. Kap119p also bound directly to a number of peptide repeat containing nucleoporins in overlay assays. In wild-type cells, TFIIS was primarily localized to the nucleus. In a strain where KAP119 has been deleted, TFIIS was mislocalized to the cytoplasm indicating that TFIIS is imported into the nucleus by Kap119p. The transport of various substrates that use other karyopherin-mediated import or export pathways was not affected in a kap119Delta strain. Hence Kap119p is a novel karyopherin that is responsible for the import of the transcription elongation factor TFIIS. PMID- 9852144 TI - Recruitment and the role of nuclear localization in polyglutamine-mediated aggregation. AB - The inherited neurodegenerative diseases caused by an expanded glutamine repeat share the pathologic feature of intranuclear aggregates or inclusions (NI). Here in cell-based studies of the spinocerebellar ataxia type-3 disease protein, ataxin-3, we address two issues central to aggregation: the role of polyglutamine in recruiting proteins into NI and the role of nuclear localization in promoting aggregation. We demonstrate that full-length ataxin-3 is readily recruited from the cytoplasm into NI seeded either by a pathologic ataxin-3 fragment or by a second unrelated glutamine-repeat disease protein, ataxin-1. Experiments with green fluorescence protein/polyglutamine fusion proteins show that a glutamine repeat is sufficient to recruit an otherwise irrelevant protein into NI, and studies of human disease tissue and a Drosophila transgenic model provide evidence that specific glutamine-repeat-containing proteins, including TATA binding protein and Eyes Absent protein, are recruited into NI in vivo. Finally, we show that nuclear localization promotes aggregation: an ataxin-3 fragment containing a nonpathologic repeat of 27 glutamines forms inclusions only when targeted to the nucleus. Our findings establish the importance of the polyglutamine domain in mediating recruitment and suggest that pathogenesis may be linked in part to the sequestering of glutamine-containing cellular proteins. In addition, we demonstrate that the nuclear environment may be critical for seeding polyglutamine aggregates. PMID- 9852145 TI - Interaction of ZPR1 with translation elongation factor-1alpha in proliferating cells. AB - The zinc finger protein ZPR1 is present in the cytoplasm of quiescent mammalian cells and translocates to the nucleus upon treatment with mitogens, including epidermal growth factor (EGF). Homologues of ZPR1 were identified in yeast and mammals. These ZPR1 proteins bind to eukaryotic translation elongation factor 1alpha (eEF-1alpha). Studies of mammalian cells demonstrated that EGF treatment induces the interaction of ZPR1 with eEF-1alpha and the redistribution of both proteins to the nucleus. In the yeast Saccharomyces cerevisiae, genetic analysis demonstrated that ZPR1 is an essential gene. Deletion analysis demonstrated that the NH2-terminal region of ZPR1 is required for normal growth and that the COOH terminal region was essential for viability in S. cerevisiae. The yeast ZPR1 protein redistributes from the cytoplasm to the nucleus in response to nutrient stimulation. Disruption of the binding of ZPR1 to eEF-1alpha by mutational analysis resulted in an accumulation of cells in the G2/M phase of cell cycle and defective growth. Reconstitution of the ZPR1 interaction with eEF-1alpha restored normal growth. We conclude that ZPR1 is essential for cell viability and that its interaction with eEF-1alpha contributes to normal cellular proliferation. PMID- 9852147 TI - Recycling of golgi-resident glycosyltransferases through the ER reveals a novel pathway and provides an explanation for nocodazole-induced Golgi scattering. AB - During microtubule depolymerization, the central, juxtanuclear Golgi apparatus scatters to multiple peripheral sites. We have tested here whether such scattering is due to a fragmentation process and subsequent outward tracking of Golgi units or if peripheral Golgi elements reform through a novel recycling pathway. To mark the Golgi in HeLa cells, we stably expressed the Golgi stack enzyme N-acetylgalactosaminyltransferase-2 (GalNAc-T2) fused to the green fluorescent protein (GFP) or to an 11-amino acid epitope, VSV-G (VSV), and the trans/TGN enzyme beta1,4-galactosyltransferase (GalT) fused to GFP. After nocodazole addition, time-lapse microscopy of GalNAc-T2-GFP and GalT-GFP revealed that scattered Golgi elements appeared abruptly and that no Golgi fragments tracked outward from the compact, juxtanuclear Golgi complex. Once formed, the scattered structures were relatively stable in fluorescence intensity for tens of minutes. During the entire process of dispersal, immunogold labeling for GalNAc T2-VSV and GalT showed that these were continuously concentrated over stacked Golgi cisternae and tubulovesicular Golgi structures similar to untreated cells, suggesting that polarized Golgi stacks reform rapidly at scattered sites. In fluorescence recovery after photobleaching over a narrow (FRAP) or wide area (FRAP-W) experiments, peripheral Golgi stacks continuously exchanged resident proteins with each other through what appeared to be an ER intermediate. That Golgi enzymes cycle through the ER was confirmed by microinjecting the dominant negative mutant of Sar1 (Sar1pdn) blocking ER export. Sar1pdn was either microinjected into untreated or nocodazole-treated cells in the presence of protein synthesis inhibitors. In both cases, this caused a gradual accumulation of GalNAc-T2-VSV in the ER. Few to no peripheral Golgi elements were seen in the nocodazole-treated cells microinjected with Sar1pdn. In conclusion, we have shown that Golgi-resident glycosylation enzymes recycle through the ER and that this novel pathway is the likely explanation for the nocodazole-induced Golgi scattering observed in interphase cells. PMID- 9852146 TI - Kinetic analysis of secretory protein traffic and characterization of golgi to plasma membrane transport intermediates in living cells. AB - Quantitative time-lapse imaging data of single cells expressing the transmembrane protein, vesicular stomatitis virus ts045 G protein fused to green fluorescent protein (VSVG-GFP), were used for kinetic modeling of protein traffic through the various compartments of the secretory pathway. A series of first order rate laws was sufficient to accurately describe VSVG-GFP transport, and provided compartment residence times and rate constants for transport into and out of the Golgi complex and delivery to the plasma membrane. For ER to Golgi transport the mean rate constant (i.e., the fraction of VSVG-GFP moved per unit of time) was 2.8% per min, for Golgi to plasma membrane transport it was 3.0% per min, and for transport from the plasma membrane to a degradative site it was 0.25% per min. Because these rate constants did not change as the concentration of VSVG-GFP in different compartments went from high (early in the experiment) to low (late in the experiment), secretory transport machinery was never saturated during the experiments. The processes of budding, translocation, and fusion of post-Golgi transport intermediates carrying VSVG- GFP to the plasma membrane were also analyzed using quantitative imaging techniques. Large pleiomorphic tubular structures, rather than small vesicles, were found to be the primary vehicles for Golgi to plasma membrane transport of VSVG-GFP. These structures budded as entire domains from the Golgi complex and underwent dynamic shape changes as they moved along microtubule tracks to the cell periphery. They carried up to 10,000 VSVG GFP molecules and had a mean life time in COS cells of 3.8 min. In addition, they fused with the plasma membrane without intersecting other membrane transport pathways in the cell. These properties suggest that the post-Golgi intermediates represent a unique transport organelle for conveying large quantities of protein cargo from the Golgi complex directly to the plasma membrane. PMID- 9852148 TI - Two functional states of the CD11b A-domain: correlations with key features of two Mn2+-complexed crystal structures. AB - In the presence of bound Mn2+, the three- dimensional structure of the ligand binding A-domain from the integrin CR3 (CD11b/CD18) is shown to exist in the "open" conformation previously described only for a crystalline Mg2+ complex. The open conformation is distinguished from the "closed" form by the solvent exposure of F302, a direct T209-Mn2+ bond, and the presence of a glutamate side chain in the MIDAS site. Approximately 10% of wild-type CD11b A-domain is present in an "active" state (binds to activation-dependent ligands, e.g., iC3b and the mAb 7E3). In the isolated domain and in the holoreceptor, the percentage of the active form can be quantitatively increased or abolished in F302W and T209A mutants, respectively. The iC3b-binding site is located on the MIDAS face and includes conformationally sensitive residues that undergo significant shifts in the open versus closed structures. We suggest that stabilization of the open structure is independent of the nature of the metal ligand and that the open conformation may represent the physiologically active form. PMID- 9852149 TI - The localization of myosin VI at the golgi complex and leading edge of fibroblasts and its phosphorylation and recruitment into membrane ruffles of A431 cells after growth factor stimulation. AB - Myosin VI is an unconventional myosin that may play a role in vesicular membrane traffic through actin rich regions of the cytoplasm in eukaryotic cells. In this study we have cloned and sequenced a cDNA encoding a chicken intestinal brush border myosin VI. Polyclonal antisera were raised to bacterially expressed fragments of this myosin VI. The affinity purified antibodies were highly specific for myosin VI by immunoblotting and immunoprecipitation and were used to study the localization of the protein by immunofluorescence and immunoelectron microscopy. It was found that in NRK and A431 cells, myosin VI was associated with both the Golgi complex and the leading, ruffling edge of the cell as well as being present in a cytosolic pool. In A431 cells in which cell surface ruffling was stimulated by EGF, myosin VI was phosphorylated and recruited into the newly formed ruffles along with ezrin and myosin V. In vitro experiments suggested that a p21-activated kinase (PAK) might be the kinase responsible for phosphorylation in the motor domain. These results strongly support a role for myosin VI in membrane traffic on secretory and endocytic pathways. PMID- 9852150 TI - Heterotrimeric kinesin II is the microtubule motor protein responsible for pigment dispersion in Xenopus melanophores. AB - Melanophores move pigment organelles (melanosomes) from the cell center to the periphery and vice-versa. These bidirectional movements require cytoplasmic microtubules and microfilaments and depend on the function of microtubule motors and a myosin. Earlier we found that melanosomes purified from Xenopus melanophores contain the plus end microtubule motor kinesin II, indicating that it may be involved in dispersion (Rogers, S.L., I.S. Tint, P.C. Fanapour, and V.I. Gelfand. 1997. Proc. Natl. Acad. Sci. USA. 94: 3720-3725). Here, we generated a dominant-negative construct encoding green fluorescent protein fused to the stalk-tail region of Xenopus kinesin-like protein 3 (Xklp3), the 95-kD motor subunit of Xenopus kinesin II, and introduced it into melanophores. Overexpression of the fusion protein inhibited pigment dispersion but had no effect on aggregation. To control for the specificity of this effect, we studied the kinesin-dependent movement of lysosomes. Neither dispersion of lysosomes in acidic conditions nor their clustering under alkaline conditions was affected by the mutant Xklp3. Furthermore, microinjection of melanophores with SUK4, a function-blocking kinesin antibody, inhibited dispersion of lysosomes but had no effect on melanosome transport. We conclude that melanosome dispersion is powered by kinesin II and not by conventional kinesin. This paper demonstrates that kinesin II moves membrane-bound organelles. PMID- 9852151 TI - Role of xklp3, a subunit of the Xenopus kinesin II heterotrimeric complex, in membrane transport between the endoplasmic reticulum and the Golgi apparatus. AB - The function of the Golgi apparatus is to modify proteins and lipids synthesized in the ER and sort them to their final destination. The steady-state size and function of the Golgi apparatus is maintained through the recycling of some components back to the ER. Several lines of evidence indicate that the spatial segregation between the ER and the Golgi apparatus as well as trafficking between these two compartments require both microtubules and motors. We have cloned and characterized a new Xenopus kinesin like protein, Xklp3, a subunit of the heterotrimeric Kinesin II. By immunofluorescence it is found in the Golgi region. A more detailed analysis by EM shows that it is associated with a subset of membranes that contain the KDEL receptor and are localized between the ER and Golgi apparatus. An association of Xklp3 with the recycling compartment is further supported by a biochemical analysis and the behavior of Xklp3 in BFA treated cells. The function of Xklp3 was analyzed by transfecting cells with a dominant-negative form lacking the motor domain. In these cells, the normal delivery of newly synthesized proteins to the Golgi apparatus is blocked. Taken together, these results indicate that Xklp3 is involved in the transport of tubular-vesicular elements between the ER and the Golgi apparatus. PMID- 9852152 TI - Centriole disassembly in vivo and its effect on centrosome structure and function in vertebrate cells. AB - Glutamylation is the major posttranslational modification of neuronal and axonemal tubulin and is restricted predominantly to centrioles in nonneuronal cells (Bobinnec, Y., M. Moudjou, J.P. Fouquet, E. Desbruyeres, B. Edde, and M. Bornens. 1998. Cell Motil. Cytoskel. 39:223-232). To investigate a possible relationship between the exceptional stability of centriole microtubules and the compartmentalization of glutamylated isoforms, we loaded HeLa cells with the monoclonal antibody GT335, which specifically reacts with polyglutamylated tubulin. The total disappearance of the centriole pair was observed after 12 h, as judged both by immunofluorescence labeling with specific antibodies and electron microscopic observation of cells after complete thick serial sectioning. Strikingly, we also observed a scattering of the pericentriolar material (PCM) within the cytoplasm and a parallel disappearance of the centrosome as a defined organelle. However, centriole disappearance was transient, as centrioles and discrete centrosomes ultimately reappeared in the cell population. During the acentriolar period, a large proportion of monopolar half-spindles or of bipolar spindles with abnormal distribution of PCM and NuMA were observed. However, as judged by a quasinormal increase in cell number, these cells likely were not blocked in mitosis. Our results suggest that a posttranslational modification of tubulin is critical for long-term stability of centriolar microtubules. They further demonstrate that in animal cells, centrioles are instrumental in organizing centrosomal components into a structurally stable organelle. PMID- 9852153 TI - Distinct mutants of retrograde intraflagellar transport (IFT) share similar morphological and molecular defects. AB - A microtubule-based transport of protein complexes, which is bidirectional and occurs between the space surrounding the basal bodies and the distal part of Chlamydomonas flagella, is referred to as intraflagellar transport (IFT). The IFT involves molecular motors and particles that consist of 17S protein complexes. To identify the function of different components of the IFT machinery, we isolated and characterized four temperature-sensitive (ts) mutants of flagellar assembly that represent the loci FLA15, FLA16, and FLA17. These mutants were selected among other ts mutants of flagellar assembly because they displayed a characteristic bulge of the flagellar membrane as a nonconditional phenotype. Each of these mutants was significantly defective for the retrograde velocity of particles and the frequency of bidirectional transport but not for the anterograde velocity of particles, as revealed by a novel method of analysis of IFT that allows tracking of single particles in a sequence of video images. Furthermore, each mutant was defective for the same four subunits of a 17S complex that was identified earlier as the IFT complex A. The occurrence of the same set of phenotypes, as the result of a mutation in any one of three loci, suggests the hypothesis that complex A is a portion of the IFT particles specifically involved in retrograde intraflagellar movement. PMID- 9852154 TI - Role of polo kinase and Mid1p in determining the site of cell division in fission yeast. AB - The fission yeast Schizosaccharomyces pombe divides symmetrically using a medial F-actin- based contractile ring to produce equal-sized daughter cells. Mutants defective in two previously described genes, mid1 and pom1, frequently divide asymmetrically. Here we present the identification of three new temperature sensitive mutants defective in localization of the division plane. All three mutants have mutations in the polo kinase gene, plo1, and show defects very similar to those of mid1 mutants in both the placement and organization of the medial ring. In both cases, ring formation is frequently initiated near the cell poles, indicating that Mid1p and Plo1p function in recruiting medial ring components to the cell center. It has been reported previously that during mitosis Mid1p becomes hyperphosphorylated and relocates from the nucleus to a medial ring. Here we show that Mid1p first forms a diffuse cortical band during spindle formation and then coalesces into a ring before anaphase. Plo1p is required for Mid1p to exit the nucleus and form a ring, and Pom1p is required for proper placement of the Mid1p ring. Upon overexpression of Plo1p, Mid1p exits the nucleus prematurely and displays a reduced mobility on gels similar to that of the hyperphosphorylated form observed previously in mitotic cells. Genetic and two-hybrid analyses suggest that Plo1p and Mid1p act in a common pathway distinct from that involving Pom1p. Plo1p localizes to the spindle pole bodies and spindles of mitotic cells and also to the medial ring at the time of its formation. Taken together, the data indicate that Plo1p plays a role in the positioning of division sites by regulating Mid1p. Given its previously known functions in mitosis and the timing of cytokinesis, Plo1p is thus implicated as a key molecule in the spatial and temporal coordination of cytokinesis with mitosis. PMID- 9852155 TI - Morphogenesis beyond cytokinetic arrest in Saccharomyces cerevisiae. AB - The budding yeast lyt1 mutation causes cell lysis. We report here that lyt1 is an allele of cdc15, a gene which encodes a protein kinase that functions late in the cell cycle. Neither cdc15-1 nor cdc15-lyt1 strains are able to septate at 37 degreesC, even though they may manage to rebud. Cells lyse after a shmoo-like projection appears at the distal pole of the daughter cell. Actin polarizes towards the distal pole but the septins remain at the mother-daughter neck. This morphogenetic response reflects entry into a new round of the cell cycle: the preference for polarization from the distal pole was lost in bud1 cdc15 double mutants; double cdc15-lyt1 cdc28-4 mutants, defective for START, did not develop apical projections and apical polarization was accompanied by DNA replication. The same phenomena were caused by mutations in the genes CDC14, DBF2, and TEM1, which are functionally related to CDC15. Apical polarization was delayed in cdc15 mutants as compared with budding in control cells and this delay was abolished in a septin mutant. Our results suggest that the delayed M/G1 transition in cdc15 mutants is due to a septin-dependent checkpoint that couples initiation of the cell cycle to the completion of cytokinesis. PMID- 9852156 TI - AIR-2: An Aurora/Ipl1-related protein kinase associated with chromosomes and midbody microtubules is required for polar body extrusion and cytokinesis in Caenorhabditis elegans embryos. AB - An emerging family of kinases related to the Drosophila Aurora and budding yeast Ipl1 proteins has been implicated in chromosome segregation and mitotic spindle formation in a number of organisms. Unlike other Aurora/Ipl1-related kinases, the Caenorhabditis elegans orthologue, AIR-2, is associated with meiotic and mitotic chromosomes. AIR-2 is initially localized to the chromosomes of the most mature prophase I-arrested oocyte residing next to the spermatheca. This localization is dependent on the presence of sperm in the spermatheca. After fertilization, AIR-2 remains associated with chromosomes during each meiotic division. However, during both meiotic anaphases, AIR-2 is present between the separating chromosomes. AIR 2 also remains associated with both extruded polar bodies. In the embryo, AIR-2 is found on metaphase chromosomes, moves to midbody microtubules at anaphase, and then persists at the cytokinesis remnant. Disruption of AIR-2 expression by RNA- mediated interference produces entire broods of one-cell embryos that have executed multiple cell cycles in the complete absence of cytokinesis. The embryos accumulate large amounts of DNA and microtubule asters. Polar bodies are not extruded, but remain in the embryo where they continue to replicate. The cytokinesis defect appears to be late in the cell cycle because transient cleavage furrows initiate at the proper location, but regress before the division is complete. Additionally, staining with a marker of midbody microtubules revealed that at least some of the components of the midbody are not well localized in the absence of AIR-2 activity. Our results suggest that during each meiotic and mitotic division, AIR-2 may coordinate the congression of metaphase chromosomes with the subsequent events of polar body extrusion and cytokinesis. PMID- 9852157 TI - Distinct roles for the p110alpha and hVPS34 phosphatidylinositol 3'-kinases in vesicular trafficking, regulation of the actin cytoskeleton, and mitogenesis. AB - We have examined the roles of the p85/ p110alpha and hVPS34 phosphatidylinositol (PI) 3'-kinases in cellular signaling using inhibitory isoform-specific antibodies. We raised anti-hVPS34 and anti-p110alpha antibodies that specifically inhibit recombinant hVPS34 and p110alpha, respectively, in vitro. We used the antibodies to study cellular processes that are sensitive to low-dose wortmannin. The antibodies had distinct effects on the actin cytoskeleton; microinjection of anti-p110alpha antibodies blocked insulin-stimulated ruffling, whereas anti hVPS34 antibodies had no effect. The antibodies also had different effects on vesicular trafficking. Microinjection of inhibitory anti-hVPS34 antibodies, but not anti-p110alpha antibodies, blocked the transit of internalized PDGF receptors to a perinuclear compartment, and disrupted the localization of the early endosomal protein EEA1. Microinjection of anti-p110alpha antibodies, and to a lesser extent anti-hVPS34 antibodies, reduced the rate of transferrin recycling in CHO cells. Surprisingly, both antibodies inhibited insulin-stimulated DNA synthesis by 80%. Injection of cells with antisense oligonucleotides derived from the hVPS34 sequence also blocked insulin-stimulated DNA synthesis, whereas scrambled oligonucleotides had no effect. Interestingly, the requirement for p110alpha and hVPS34 occurred at different times during the G1-S transition. Our data suggest that different PI 3'-kinases play distinct regulatory roles in the cell, and document an unexpected role for hVPS34 during insulin-stimulated mitogenesis. PMID- 9852158 TI - Calcium and protein kinase C regulate the actin cytoskeleton in the synaptic terminal of retinal bipolar cells. AB - The organization of filamentous actin (F-actin) in the synaptic pedicle of depolarizing bipolar cells from the goldfish retina was studied using fluorescently labeled phalloidin. The amount of F-actin in the synaptic pedicle relative to the cell body increased from a ratio of 1.6 +/- 0.1 in the dark to 2.1 +/- 0.1 after exposure to light. Light also caused the retraction of spinules and processes elaborated by the synaptic pedicle in the dark. Isolated bipolar cells were used to characterize the factors affecting the actin cytoskeleton. When the electrical effect of light was mimicked by depolarization in 50 mM K+, the actin network in the synaptic pedicle extended up to 2.5 micrometer from the plasma membrane. Formation of F-actin occurred on the time scale of minutes and required Ca2+ influx through L-type Ca2+ channels. Phorbol esters that activate protein kinase C (PKC) accelerated growth of F-actin. Agents that inhibit PKC hindered F-actin growth in response to Ca2+ influx and accelerated F-actin breakdown on removal of Ca2+. To test whether activity-dependent changes in the organization of F-actin might regulate exocytosis or endocytosis, vesicles were labeled with the fluorescent membrane marker FM1-43. Disruption of F-actin with cytochalasin D did not affect the continuous cycle of exocytosis and endocytosis that was stimulated by maintained depolarization, nor the spatial distribution of recycled vesicles within the synaptic terminal. We suggest that the actions of Ca2+ and PKC on the organization of F-actin regulate the morphology of the synaptic pedicle under varying light conditions. PMID- 9852159 TI - Neurite fasciculation mediated by complexes of axonin-1 and Ng cell adhesion molecule. AB - Neural cell adhesion molecules composed of immunoglobulin and fibronectin type III-like domains have been implicated in cell adhesion, neurite outgrowth, and fasciculation. Axonin-1 and Ng cell adhesion molecule (NgCAM), two molecules with predominantly axonal expression exhibit homophilic interactions across the extracellular space (axonin- 1/axonin-1 and NgCAM/NgCAM) and a heterophilic interaction (axonin-1-NgCAM) that occurs exclusively in the plane of the same membrane (cis-interaction). Using domain deletion mutants we localized the NgCAM homophilic binding in the Ig domains 1-4 whereas heterophilic binding to axonin-1 was localized in the Ig domains 2-4 and the third FnIII domain. The NgCAM-NgCAM interaction could be established simultaneously with the axonin-1-NgCAM interaction. In contrast, the axonin-1-NgCAM interaction excluded axonin-1/axonin 1 binding. These results and the examination of the coclustering of axonin-1 and NgCAM at cell contacts, suggest that intercellular contact is mediated by a symmetric axonin-12/NgCAM2 tetramer, in which homophilic NgCAM binding across the extracellular space occurs simultaneously with a cis-heterophilic interaction of axonin-1 and NgCAM. The enhanced neurite fasciculation after overexpression of NgCAM by adenoviral vectors indicates that NgCAM is the limiting component for the formation of the axonin-12/NgCAM2 complexes and, thus, neurite fasciculation in DRG neurons. PMID- 9852160 TI - p53 is essential for developmental neuron death as regulated by the TrkA and p75 neurotrophin receptors. AB - Naturally occurring sympathetic neuron death is the result of two apoptotic signaling events: one normally suppressed by NGF/TrkA survival signals, and a second activated by the p75 neurotrophin receptor. Here we demonstrate that the p53 tumor suppressor protein, likely as induced by the MEKK-JNK pathway, is an essential component of both of these apoptotic signaling cascades. In cultured neonatal sympathetic neurons, p53 protein levels are elevated in response to both NGF withdrawal and p75NTR activation. NGF withdrawal also results in elevation of a known p53 target, the apoptotic protein Bax. Functional ablation of p53 using the adenovirus E1B55K protein inhibits neuronal apoptosis as induced by either NGF withdrawal or p75 activation. Direct stimulation of the MEKK-JNK pathway using activated MEKK1 has similar effects; p53 and Bax are increased and the subsequent neuronal apoptosis can be rescued by E1B55K. Expression of p53 in sympathetic neurons indicates that p53 functions downstream of JNK and upstream of Bax. Finally, when p53 levels are reduced or absent in p53+/- or p53-/- mice, naturally occurring sympathetic neuron death is inhibited. Thus, p53 is an essential common component of two receptor-mediated signal transduction cascades that converge on the MEKK-JNK pathway to regulate the developmental death of sympathetic neurons. PMID- 9852161 TI - Overexpression of VEGF in testis and epididymis causes infertility in transgenic mice: evidence for nonendothelial targets for VEGF. AB - Vascular endothelial growth factor (VEGF) is a key regulator of endothelial growth and permeability. However, VEGF may also target nonendothelial cells, as VEGF receptors and responsiveness have been detected for example in monocytes, and high concentrations of VEGF have been reported in human semen. In this work we present evidence that overexpression of VEGF in the testis and epididymis of transgenic mice under the mouse mammary tumor virus (MMTV) LTR promoter causes infertility. The testes of the transgenic mice exhibited spermatogenic arrest and increased capillary density. The ductus epididymidis was dilated, containing areas of epithelial hyperplasia. The number of subepithelial capillaries in the epididymis was also increased and these vessels were highly permeable as judged by the detection of extravasated fibrinogen products. Intriguingly, the expression of VEGF receptor-1 (VEGFR-1) was detected in certain spermatogenic cells in addition to vascular endothelium, and both VEGFR-1 and VEGFR-2 were also found in the Leydig cells of the testis. The infertility of the MMTV-VEGF male mice could thus result from VEGF acting on both endothelial and nonendothelial cells of the male genital tract. Taken together, these findings suggest that the VEGF transgene has nonendothelial target cells in the testis and that VEGF may regulate male fertility. PMID- 9852162 TI - Roles for laminin in embryogenesis: exencephaly, syndactyly, and placentopathy in mice lacking the laminin alpha5 chain. AB - Laminins are the major noncollagenous glycoproteins of all basal laminae (BLs). They are alpha/beta/gamma heterotrimers assembled from 10 known chains, and they subserve both structural and signaling roles. Previously described mutations in laminin chain genes result in diverse disorders that are manifested postnatally and therefore provide little insight into laminin's roles in embryonic development. Here, we show that the laminin alpha5 chain is required during embryogenesis. The alpha5 chain is present in virtually all BLs of early somite stage embryos and then becomes restricted to specific BLs as development proceeds, including those of the surface ectoderm and placental vasculature. BLs that lose alpha5 retain or acquire other alpha chains. Embryos lacking laminin alpha5 die late in embryogenesis. They exhibit multiple developmental defects, including failure of anterior neural tube closure (exencephaly), failure of digit septation (syndactyly), and dysmorphogenesis of the placental labyrinth. These defects are all attributable to defects in BLs that are alpha5 positive in controls and that appear ultrastructurally abnormal in its absence. Other laminin alpha chains accumulate in these BLs, but this compensation is apparently functionally inadequate. Our results identify new roles for laminins and BLs in diverse developmental processes. PMID- 9852163 TI - Gap junction-mediated cell-cell communication modulates mouse neural crest migration. AB - Previous studies showed that conotruncal heart malformations can arise with the increase or decrease in alpha1 connexin function in neural crest cells. To elucidate the possible basis for the quantitative requirement for alpha1 connexin gap junctions in cardiac development, a neural crest outgrowth culture system was used to examine migration of neural crest cells derived from CMV43 transgenic embryos overexpressing alpha1 connexins, and from alpha1 connexin knockout (KO) mice and FC transgenic mice expressing a dominant-negative alpha1 connexin fusion protein. These studies showed that the migration rate of cardiac neural crest was increased in the CMV43 embryos, but decreased in the FC transgenic and alpha1 connexin KO embryos. Migration changes occurred in step with connexin gene or transgene dosage in the homozygous vs. hemizygous alpha1 connexin KO and CMV43 embryos, respectively. Dye coupling analysis in neural crest cells in the outgrowth cultures and also in the living embryos showed an elevation of gap junction communication in the CMV43 transgenic mice, while a reduction was observed in the FC transgenic and alpha1 connexin KO mice. Further analysis using oleamide to downregulate gap junction communication in nontransgenic outgrowth cultures showed that this independent method of reducing gap junction communication in cardiac crest cells also resulted in a reduction in the rate of crest migration. To determine the possible relevance of these findings to neural crest migration in vivo, a lacZ transgene was used to visualize the distribution of cardiac neural crest cells in the outflow tract. These studies showed more lacZ-positive cells in the outflow septum in the CMV43 transgenic mice, while a reduction was observed in the alpha1 connexin KO mice. Surprisingly, this was accompanied by cell proliferation changes, not in the cardiac neural crest cells, but in the myocardium- an elevation in the CMV43 mice vs. a reduction in the alpha1 connexin KO mice. The latter observation suggests that cardiac neural crest cells may have a role in modulating growth and development of non-neural crest- derived tissues. Overall, these findings suggest that gap junction communication mediated by alpha1 connexins plays an important role in cardiac neural crest migration. Furthermore, they indicate that cardiac neural crest perturbation is the likely underlying cause for heart defects in mice with the gain or loss of alpha1 connexin function. PMID- 9852164 TI - Cellular interaction of integrin alpha3beta1 with laminin 5 promotes gap junctional communication. AB - Wounding of skin activates epidermal cell migration over exposed dermal collagen and fibronectin and over laminin 5 secreted into the provisional basement membrane. Gap junctional intercellular communication (GJIC) has been proposed to integrate the individual motile cells into a synchronized colony. We found that outgrowths of human keratinocytes in wounds or epibole cultures display parallel changes in the expression of laminin 5, integrin alpha3beta1, E-cadherin, and the gap junctional protein connexin 43. Adhesion of keratinocytes on laminin 5, collagen, and fibronectin was found to differentially regulate GJIC. When keratinocytes were adhered on laminin 5, both structural (assembly of connexin 43 in gap junctions) and functional (dye transfer) assays showed a two- to threefold increase compared with collagen and five- to eightfold over fibronectin. Based on studies with immobilized integrin antibody and integrin-transfected Chinese hamster ovary cells, the interaction of integrin alpha3beta1 with laminin 5 was sufficient to promote GJIC. Mapping of intermediate steps in the pathway linking alpha3beta1-laminin 5 interactions to GJIC indicated that protein trafficking and Rho signaling were both required. We suggest that adhesion of epithelial cells to laminin 5 in the basement membrane via alpha3beta1 promotes GJIC that integrates individual cells into synchronized epiboles. PMID- 9852165 TI - Release of cAMP gating by the alpha6beta4 integrin stimulates lamellae formation and the chemotactic migration of invasive carcinoma cells. AB - The alpha6beta4 integrin promotes carcinoma in-vasion by its activation of a phosphoinositide 3-OH (PI3-K) signaling pathway (Shaw, L.M., I. Rabinovitz, H.H. F. Wang, A. Toker, and A.M. Mercurio. Cell. 91: 949-960). We demonstrate here using MDA-MB-435 breast carcinoma cells that alpha6beta4 stimulates chemotactic migration, a key component of invasion, but that it has no influence on haptotaxis. Stimulation of chemotaxis by alpha6beta4 expression was observed in response to either lysophosphatidic acid (LPA) or fibroblast conditioned medium. Moreover, the LPA-dependent formation of lamellae in these cells is dependent upon alpha6beta4 expression. Both lamellae formation and chemotactic migration are inhibited or "gated" by cAMP and our results reveal that a critical function of alpha6beta4 is to suppress the intracellular cAMP concentration by increasing the activity of a rolipram-sensitive, cAMP-specific phosphodiesterase (PDE). This PDE activity is essential for lamellae formation, chemotactic migration and invasion based on data obtained with PDE inhibitors. Although PI3-K and cAMP specific PDE activities are both required to promote lamellae formation and chemotactic migration, our data indicate that they are components of distinct signaling pathways. The essence of our findings is that alpha6beta4 stimulates the chemotactic migration of carcinoma cells through its ability to influence key signaling events that underlie this critical component of carcinoma invasion. PMID- 9852167 TI - Glycosylation and palmitoylation are not required for the formation of the X linked cone opsin visual pigments. AB - PURPOSE: This study was designed to test whether palmitoylation and glycosylation are required for the formation of the green opsin visual pigment. METHODS: Stable cell lines were established by transfecting EBNA-293 cells with a pMEP4ss recombinant plasmid containing wild-type bovine rhodopsin or wild-type or mutant (N32S) green opsin cDNA molecules that included a tag for the eight amino acid residues located at the C-terminus of rhodopsin. The opsins were induced by addition of CdCl2 into the medium and then reconstituted with 11-cis-retinal. The reconstituted opsins were purified by immunoaffinity chromatography, then analyzed by difference spectra, and by binding 35S-GTP in the presence of bovine transducin. Non-reconstituted opsins were analyzed by Western blotting and by pulse-labeling with 3H-palmitic acid followed by immunoprecipitation. RESULTS: Elimination of glycosylation by mutagenesis of the N-linked glycosylation site did not impair the ability of the resulting cone opsin to absorb light at the appropriate wavelength nor to activate transducin. Furthermore, as judged by pulse-labeling with 3H-palmitic acid and immunoprecipitation and by gas chromatography-mass spectroscopy, the wild type green opsin differs from rhodopsin by not being palmitoylated. CONCLUSIONS: Glycosylation and palmitoylation are not required for the formation of cone opsin visual pigments. For the previously described green opsin C203R mutation, disruption of folding and transport, rather than altered glycosylation is sufficient to explain the associated color vision deficiency. PMID- 9852166 TI - The cloning of GRK7, a candidate cone opsin kinase, from cone- and rod-dominant mammalian retinas. AB - PURPOSE: Desensitization in the rod cell of the mammalian retina is initiated when light-activated rhodopsin is phosphorylated by the G protein-coupled receptor kinase (GRK), GRK1, often referred to as rhodopsin kinase. A distinct kinase that specifically phosphorylates cone opsins in a similar manner has not been identified in mammals. To determine the existence of a cone opsin kinase, RNA from the retinas of cone- and rod-dominant mammals was analyzed by PCR. METHODS: RNA prepared from the retinas of two cone-dominant mammals, the thirteen lined ground squirrel and the eastern chipmunk, and a rod-dominant mammal, the pig, was used to clone a new GRK family member by RT-PCR. The tissue distribution and localization of the kinase in retina were determined by Northern blot hybridization and in situ hybridization. The protein encoded by this cDNA was expressed in human embryonic kidney-293 (HEK-293) cells and compared with bovine GRK1 for its ability to phosphorylate bovine rhodopsin and to undergo autophosphorylation. RESULTS: The cDNA cloned from ground squirrel contains an open reading frame encoding a 548 amino-acid protein. Sequence analysis indicates that this protein is orthologous to GRK7 recently cloned from O. latipes, the medaka fish. Partial cDNA fragments of GRK7 were also cloned from RNA prepared from eastern chipmunk and pig retinas. In situ hybridization demonstrated widespread labeling in the photoreceptor layer of the ground squirrel retina, consistent with expression in cones. Recombinant ground squirrel GRK7 phosphorylates bovine rhodopsin in a light-dependent manner and can be autophosphorylated, similar to bovine GRK1. CONCLUSIONS: These results indicate that cone- and rod-dominant mammals both express GRK7. The presence of this kinase in cones in the ground squirrel and its ability to phosphorylate rhodopsin suggests that it could function in cone cells as a cone opsin kinase. PMID- 9852168 TI - Primary reconstruction of the ala nasi in cleft Lip. AB - Displacement of the lateral crus of the alar cartilage of the nose in cleft lip is demonstrable in diagrams showing the pathological anatomy of this deformity. An account is given of our method used for several years, whereby the lateral limb is returned to a more natural position, alar convexity is restored, and the angle between the limbs is corrected. Two flaps taken from the inside surface of the nostril are alternated. The caudal one is a full-thickness flap composed of vestibular skin, while the cranial flap comprises the cartilage and the vestibular mucosa; the lateral limb is left attached to the mucosa. Alternation does not require any interruption of cartilage continuity, so there is no risk of surgical damage to the perichondrium and cartilage in young subjects. The two limbs (particularly the lateral one) are positioned in such a way that their normal growth can be expected, as shown by the aesthetic results on follow-up after a period of many years. This method is a primary reconstruction technique of the ala nasi, applied at the same time as the lip repair and compatible with current methods. PMID- 9852169 TI - Lipoplasty-another plea for safety! PMID- 9852170 TI - Liposculpture with ultrasound: biomedical considerations. AB - The authors, after a careful review of the scientific literature and on the basis of biophysical concepts with their own experience, conclude that high-intensity ultrasounds (US) are responsible for some biological lesions, which are partly unknown. US can cause burns and skin necrosis; thus, our employment of this technique must be correct, justified, and practiced very carefully and shrewdly and its advantages versus traditional liposuction must be reviewed. PMID- 9852171 TI - Tumescent abdominoplasty: an ambulatory office procedure. AB - Abdominoplasty has traditionally been a procedure that has been performed in a hospital setting, usually with an overnight stay, particularly when full rectus plication has been part of the operative procedure. With the development of office-based operating rooms and improved office anesthesia, the ability to perform more extensive surgery outside the hospital is now possible. Additionally, the cost savings of office-based surgery compared with noncontrolled hospital charges makes it much easier for patients to plan the exact charges that they will encounter. Experience with the tumescent technique for liposuction has demonstrated to the plastic surgeon that extensive surgery can be performed with minimal postoperative discomfort. The tumescent technique has also been used to perform miniabdominoplasty [1]. By using this technique in patients undergoing full abdominoplasty, this procedure can be performed in the office with minimal blood loss and almost no need for pain management in the early postoperative period, and the patients can be discharged home within 3 h of the completion of the procedure. PMID- 9852172 TI - Liposuction of the kneecap area under tourniquet: a superdry procedure. AB - In this paper, we report our experience and describe the technique of liposuction under tourniquet. This technique facilitates liposuction in the kneecap area and makes it bloodless. The approach described is effective and necessitates a relatively brief intraoperative and postoperative period, with inherently less morbidity. We obtained satisfactory results using this simple procedure. PMID- 9852173 TI - Life satisfaction, self-esteem, and body image: a psychosocial evaluation of aesthetic and reconstructive surgery candidates. AB - Psychology should be an important part of a plastic surgery procedure in order to buoy up the patient so that he gains a consequent relief of psychiatric and social problems. It is a fact that reconstructive surgery patients also seek psychological stability and a better appearance that is acceptable to society. While the life satisfaction, self-esteem, and body image of the individual who seeks aesthetic plastic surgery has been a special concern in the plastic surgery literature, data about reconstructive surgery patients are usually lacking. To understand the factors influencing the aim of an individual seeking plastic surgery, this prospective controlled clinical study was designed. We hypothesized that they should have lower life satisfaction and self-esteem and more distorted body images than the "normal" population. Three hundred individuals were included in the study. Both the aesthetic surgery group (AG) and the reconstructive surgery group (RG) consist of 100 (n = 100) subjects. Individuals chosen randomly from the total population seen in the 12-month period were included regardless of the type of procedure, body region, or type of trauma as well as whether or not their wish for surgery was justified by the staff. The third group, the control group (CG), also consists of 100 (n = 100) subjects and was chosen from the "normal" population. All 300 subjects were asked to complete four scales: the Socio-Demographic Questionnaire, Life-Satisfaction Index (LSI), Self-Esteem Inventory (SEI), and Body-Image Inventory (BII). Results were collected and compared. The LSI results for the two surgical groups were found to be similar to those for the CG, with the RG demonstrating the lowest level. For the SEI there was a significant difference between the two surgical groups, being in favor of the AG, and a greater difference was noted between the CG and the RG. BII results showed no significant difference among the three groups. As a result, although there may be some, a regular person who seeks an aesthetic procedure should not be considered a psychologically disturbed individual at face value and each case should be evaluated individually in the preoperative consultation. Similarly, individuals who present for reconstructive procedures and wish further aesthetic refinements should be evaluated and treated exactly as pure aesthetic candidates. PMID- 9852174 TI - Motivation for rhinoplasty: changes in 5970 cases, in three groups, 1964 to 1997. AB - The motivational factors were divided into three categories: (1) independent decision, (2) observation of previous patients, and (3) external influences. The results of the percentage breakdown of the patients in our present study are compared with previous results are presented in 1983 in this journal. Once again, the independent decision of the patient was the most important factor in deciding to have cosmetic rhinoplasty, followed by their observation of results in other patients. These two factors have been the motivation in approximately 80% of all the patients in all three studies. The age breakdown of cosmetic rhinoplasty patients shows that after the age of 21, as patients get older, a smaller and smaller percentage of patients was motivated for surgery by independent decision alone. These older patients need external influences to motivate them to have surgery. In most cases, the desire for surgery had smoldered in them since adolescence, and they needed an external "OK" to have it done. PMID- 9852175 TI - "Flip-flap" mastopexy. AB - The "flip-flap" mastopexy modifies the McKissock vertical bipedicle design by creating a wide superiorly based flap of breast tissue deep to the vertical bipedicle. Transposition of this flap up and beneath the upper breast, with suture anchoring to the pectoralis fascia at the level of the second rib, restores upper breast fullness, decreases mastopexy-wrecking lower breast bulk, and provides a pleasant forward thrust of the nipple-areolar complex. The "flip flap" is effective for improving the long-term aesthetic outcome for both reduction mammaplasty and mastopexies of moderate- to full-sized breasts. PMID- 9852176 TI - A variant of Tietze's syndrome occurring after reconstructive breast surgery. AB - Costosternal chondrodynia (Tietze's syndrome variant) should be considered in those rare patients who present with severe breast pain several months after reconstructive breast surgery. The authors treated 25 cases, 3 of which are discussed in detail, where the origin of the pain was traced to the costosternal cartilages. The differential diagnosis and management of this hitherto undescribed problem after breast surgery are discussed. PMID- 9852177 TI - Asian blepharoplasty with a short-pulsed contact Nd-Yag laser: limited-incision resectable laser double fold with internal medial and lateral functional epicanthoplasty. AB - The double-fold operation is the most common aesthetic procedure in Asia, but complications such as gross asymmetry, loss of folds, and scars on the medial canthal region and upper eyelid are relatively common. To prevent these complications, caused by displacement of fixation sutures secondary to excessive edema or hematoma, we use fine fiber-delivered contact surgical Nd-Yag (Japan SLT) laser for incision, excision, coagulation, dissection, ablation, and fusion of the soft tissue. Comparing this laser treatment to conventional electrocautery techniques or to CO2 laser blepharoplasty, this contact short-pulsed Nd-Yag laser gives us a tactile sensation, providing a more precise, controlled surgical procedure without lateral thermal damage, and it is possible to operate through a small incision. A long-term follow-up study shows that there was no gross asymmetry, loss of folds, or visible or hypertrophic scars on the upper eyelid and medial canthal regions. PMID- 9852178 TI - Tissue adhesives and applications in plastic and reconstructive surgery. AB - The search for the perfect surgical "glue" is ongoing. Throughout the centuries, surgeons and scientists have sought an ideal sealant and adhesive: one that is safe, biologically compatible, effective, and affordable. A tissue sealant with these attributes would have unlimited applications in the field of plastic and reconstructive surgery. Simple wound closure, sealing of cavities, attachment of graft and flaps, and decreasing of bleeding, bruising, and edema are problems that all aesthetic and reconstructive surgeons face every day in their practice. PMID- 9852179 TI - Bioplastique as a complement in conventional plastic surgery. AB - This research presents reports of cases where a biocompatible and alloplastic biomaterial-Bioplastique-was used, associated with conventional plastic surgery or as a complement to it, with the aim of achieving a better final aesthetic result. Four cases are presented where Bioplastique was used in association with rhytidoplasty, rhinoplasty, and other surgical techniques. This material has shown itself to be appropriate to complement surgery; achieving a final result which would not be possible without any resort to a complement or any other hard procedure by the surgeon and is not more traumatic for the patient. PMID- 9852181 TI - FORUM: Enforcement Lessons from the Vienna Ubahn. AB - / This paper combines a review of recent publications on the effectiveness of environmental enforcement in the United States with new data to address the question of what type of enforcement activity is most productive. Using data on 39 state National Pollutant Discharge Elimination System (NPDES) programs, the measures of effort and cost typically applied to environmental enforcement and inspection activities were tested for relationships with compliance outcomes. No statistically significant relationships were found. In the absence of any improving trend in traditional measures of compliance, this lack suggests all enforcement systems presently used by states are failing. To work, an environmental enforcement system needs to include maintenance and restoration of compliance, real deterrence, mobilization of public opinion, minimal obtrusiveness, conformity with legal search requirements, cost-effectiveness for all parties, effective primary role for skilled inspectors, self-monitoring, compatibility with environmental management systems (e.g., ISO 14000), environmental auditing, robustness in the face of changing strategies by permittees, segregation of technical assistance from enforcement, team orientation, adaptability to multimedia, and, most difficult of all, disconnection from today's timely, appropriate, proportionate standard. These goals can be achieved through a proposed new environmental enforcement approach built on infrequent, random, but thorough inspections leading to fully documented enforcement actions resulting in high penalties. The enforcement system used by the Vienna Ubahn, or subway, can serve as a model for this type of environmental enforcement system. KEY WORDS: Enforcement; Deterrence; Environmental water quality; Discharge elimination systems PMID- 9852180 TI - A triangular suture to simplify reduction mammaplasty. AB - A technique to facilitate correct preoperative marking for an en bloc resection in superiorly based mammaplasty is presented. A temporary triangular suture is placed to control the estimated resection lines for the removal of the medial and lateral tissue block. The described technique helps to avoid the formation of medial and lateral dog ears and the unpleasant and visible scars that result. PMID- 9852182 TI - Waste Management in the European Union: National Self-Sufficiency and Harmonization at the Expense of Economic Efficiency? AB - / Since 1975, the European Union (EU) has developed an impressive body of waste management legislation, although, according to the principle of subsidiarity, responsibilities belong to the most local level at which the tasks can be carried out effectively. Using economic theory, this paper examines several factors that influence the efficient distribution of regulatory authority among the EU and the member states. We specifically analyze the role of harmonization that, in our opinion, will often lead to an overcentralization. Within this context, a second issue is becoming more important, as ongoing discussions show. If no consensus can be reached about harmonization, should member states be allowed to stop cross border shipments of waste, or should the EU strive for a common market for waste? We take the position that most objections against waste shipments are not convincing, especially if the member states implement minimum standards for landfills and waste processing facilities and common information and control systems. Competition between different national regulations within adequate constraints that, e.g., control transboundary externalities leads to more efficient waste management structures in Europe than national self-sufficiency or centralized decision-making at the EU level. KEY WORDS: Waste management; European Union; Subsidiarity; Institutional competition; Harmonization; Cross border shipments of waste PMID- 9852183 TI - Curriculum Development in Environmental Science: A Case Study on Paradigm and Institutions. AB - / The case of "Environment and Development" at Leiden University, the Netherlands, offers an example of developing a new environmental science curriculum in a conservative, disciplines-oriented university context. The core of this history is the long-term struggle of environmental science to evolve from the level of doing applied interdisciplinary studies and establish itself as a distinct body of knowledge with its own theory level, i.e., a discipline of its own. The struggle itself as well as its final outcome, a "bidisciplinary" curriculum in which both environmental science and one social science are expressed as disciplines (hence not environmental science as a mere "field of application") may be of value in other "classical" universities, too. KEY WORDS: Environmental science; Curriculum; Interdisciplinarity; Universities PMID- 9852184 TI - PROFILE: Urban Stream Rehabilitation: A Design and Construction Case Study. AB - / This paper describes the fundamental design features, and construction methods and sequence, of a rehabilitation project on a small suburban creek in Moscow, Idaho, USA. A meandering channel pattern was reestablished for approximately 280 m of straightened, dredged channel, a new floodplain was excavated, and the new riparian zone was replanted. The new stream channel was sized to accommodate an estimated natural bankfull discharge ( approximately 5.6 cms), and floodplain design attempted to match the conveyance of the old enlarged channel (14-20 cms). The project was coordinated by a local nonprofit environmental organization, and the design and construction were tailored to donated materials and a largely volunteer labor force. A high-magnitude flood event (ca. 50-year recurrence interval) six months after construction had no significant impact on the newly constructed channel and revetments, but underscored the need for important detailing of the structures. The use of volunteer labor, while entailing certain benefits, complicates project planning and construction. The most general lesson learned from this project is that sponsoring agencies and clients need to be informed of the many steps and sequencing of properly constructed, complex stream rehabilitation projects as well as the high time and cost requirements for these tasks. KEY WORDS: Stream corridor restoration; Channel design; Streambank revetments PMID- 9852185 TI - Establishing a Regional Monitoring Strategy: The Pacific Northwest Forest Plan. AB - / This paper identifies lessons learned and issues raised during the development of an ecosystem monitoring strategy intended to support the Northwest Forest Plan. Adaptive ecosystem management, which requires monitoring as essential feedback to management, recognizes that action is necessary or appropriate, although knowledge may be imperfect. We suggest that this principle be explicitly acknowledged in the design of monitoring programs, and we coin the term adaptive monitoring design. Adaptive monitoring design is an iterative process that refines the specifications for monitoring over time as a result of experience in implementing a monitoring program, assessing results, and interacting with users. An adaptive design therefore facilitates ecosystem management. We also discuss lessons of temporal and spatial scales raised by the consideration of a design for ecosystem management. Three additional issues-integration of information from different sources, institutional infrastructure, and the roles of individuals working in an interagency setting-are also identified, but not developed in detail. KEY WORDS: Northwest Forest Plan; Monitoring; Ecosystem management; Adaptive management; Adaptive monitoring PMID- 9852186 TI - Computer Simulation Modeling for Recreation Management: A Study on Carriage Road Use in Acadia National Park, Maine, USA. AB - / The number of visits to outdoor recreation areas has increased dramatically in the last three decades, leading managers and researchers to wonder if there is a limit to the amount of use a resource such as a park can accommodate. One of the difficulties in addressing this carrying capacity-related question has been the complex nature of visitor travel patterns on often extensive networks of roads and trails. Systematic direct observation is often impractical and anecdotal information is usually inadequate. This study explores the utility of computer simulation as a tool for describing visitor travel by building a dynamic model of visitor travel on the carriage roads of Acadia National Park, Maine, USA. The simulation model uses empirical inputs such as travel routes and travel speeds to generate simulated recreation days on the carriage roads. Data on persons-per viewscape (PPV) conditions were then gathered from multiple model runs and incorporated into the National Park Service's visitor experience and resource protection (VERP) planning process. Results show that PPV conditions under present-day use levels do not violate proposed standards of quality. Results also show likely PPV conditions under scenarios of increasing use and in different areas within the carriage road system. Goodness-of-fit validity tests indicate the model is an accurate representation of the actual system. The findings of this study suggest that computer simulation is useful for estimating current carrying capacity conditions, predicting future conditions, and guiding related research. KEY WORDS: Computer simulation modeling; Carrying capacity; Recreation management; Acadia National Park PMID- 9852187 TI - Towards a Sustainable Natural Resource Management of Tribal Communities: Findings from a Study of Swidden and Wetland Cultivation in Remote Hill Regions of Eastern India. AB - / Environmental degradation in many hill forest regions of Asia, inhabited by indigenous/tribal communities is growing at an unprecedented rate. The case of Orissa State in eastern India is no exception. The government is of the view that the local population is responsible for forest degradation as they practice swidden cultivation and forest gathering indiscriminately to sustain their livelihoods. Based on economic merit alone, the government has undertaken some policy initiatives. Such initiatives that are meant to stop swidden cultivation have not been successful. This study recommends an integrated framework for developing a sustainable natural resource management practice for tribal communities. The framework has taken into consideration both economic and noneconomic factors in evaluating various alternatives. Furthermore, it has been applied to two tribal communities in Orissa (Juang and Saora). Based on an economic analysis comparing returns from swidden as well as wetland cultivation, it is observed that where forest degradation is not serious, wetland cultivation does not have a significant economic merit vis-a-vis swidden cultivation. However, in view of the long-term sustainability issue within ecological limits, swidden cultivation may be phased out in favor of wetland cultivation with an appropriate transition period. During this transition, government must adopt suitable policy initiatives to provide tribals tenurial rights to land, help financially in creating settled lands through terracing, introduce certain improved agroforestry techniques and train tribal people in other income generating activities. Furthermore, all such interventions made by the government should have a strong sociocultural component in order to attract the tribal people to give up swidden cultivation. PMID- 9852188 TI - RESEARCH: Effects of Recent Volcanic Eruptions on Aquatic Habitat in the Drift River, Alaska, USA: Implications at Other Cook Inlet Region Volcanoes. AB - / Numerous drainages supporting productive salmon habitat are surrounded by active volcanoes on the west side of Cook Inlet in south-central Alaska. Eruptions have caused massive quantities of flowing water and sediment to enter the river channels emanating from glaciers and snowfields on these volcanoes. Extensive damage to riparian and aquatic habitat has commonly resulted, and benthic macroinvertebrate and salmonid communities can be affected. Because of the economic importance of Alaska's fisheries, detrimental effects on salmonid habitat can have significant economic implications. The Drift River drains glaciers on the northern and eastern flanks of Redoubt Volcano. During and following eruptions in 1989-1990, severe physical disturbances to the habitat features of the river adversely affected the fishery. Frequent eruptions at other Cook Inlet region volcanoes exemplify the potential effects of volcanic activity on Alaska's important commercial, sport, and subsistence fisheries. Few studies have documented the recovery of aquatic habitat following volcanic eruptions. The eruptions of Redoubt Volcano in 1989-1990 offered an opportunity to examine the recovery of the macroinvertebrate community. Macroinvertebrate community composition and structure in the Drift River were similar in both undisturbed and recently disturbed sites. Additionally, macroinvertebrate samples from sites in nearby undisturbed streams were highly similar to those from some Drift River sites. This similarity and the agreement between the Drift River macroinvertebrate community composition and that predicted by a qualitative model of typical macroinvertebrate communities in glacier-fed rivers indicate that the Drift River macroinvertebrate community is recovering five years after the disturbances associated with the most recent eruptions of Redoubt Volcano. KEY WORDS: Aquatic habitat; Volcanoes; Lahars; Lahar-runout flows; Macroinvertebrates; Community structure; Community composition; Taxonomic similarity PMID- 9852189 TI - The Effects of Boat Docks on Eelgrass Beds in Coastal Waters of Massachusetts. AB - / The effects of docks on eelgrass beds were measured using plant population characteristics (shoot density, canopy height, and growth rates), light levels, and an assessment of eelgrass bed quality in Massachusetts estuaries. Eelgrass populations were impacted under and directly adjacent to docks, as shown by depressed shoot density and canopy structure. Severe impacts can cause fragmentation of eelgrass beds, thus contributing to large-scale declines in estuaries such as Waquoit Bay. Impacts were fewer under docks supported by piers than under floating docks, and taller piers resulted in fewer impacts. Based on this investigation, we conclude that docks should be greater than 3 m in height above the bottom in areas with tidal ranges less than 1 m to allow enough light to sustain eelgrass beds under the docks. In addition to dock height, orientation and width were also found to be important factors affecting eelgrass. Narrow docks with a north-south orientation can best ensure the long-term survival of eelgrass under and near the dock. KEY WORDS: Docks; Shade; Eelgrass beds; Zostera marina; Seagrass PMID- 9852190 TI - Biological Control of Water Hyacinth Under Conditions of Maintenance Management: Can Herbicides and Insects Be Integrated? AB - / We hypothesized that repeated herbicidal (maintenance) control of water hyacinth infestations in Florida suppressed biological control agent populations, especially the weevils Neochetina eichhorniae and N. bruchi. We therefore sampled water hyacinth and weevil populations at 54 sites distributed statewide. Half were under maintenance control, half were not treated with herbicides. General site conditions were assessed, demographic data were collected on weevil and plant populations, the reproductive condition of the weevils was determined, and plant nutrient and proximate composition of water hyacinth leaves were analyzed. Water hyacinth infestations under maintenance control were minimal when compared to unmanaged sites. Likewise, on a population basis, all weevil cohorts were much lower due to the paucity of plants. Plants at unmanaged sites, where weevil intensities were much higher, suffered high levels of stress and showed low growth potential. Lower percentages of the female weevils were reproductive at unmanaged sites when compared to managed sites, so densities of reproductives and immatures were similar at both site types. Reproductive status of the weevils improved with increased plant quality. Plant quality, in turn, declined as stresses arising from weevil feeding increased. Plant quality was positively correlated with plant growth potential and flower production. Thus, maintenance control improved plant nutritive quality thereby inducing reproductive vigor of the weevils, but ensuring plant regrowth and the need for future control. This suggests that biological and herbicidal controls should be integrated, using herbicides to maintain water hyacinth infestations below management thresholds but in a manner that conserves biological control agent populations. This approach would lead to improved plant nutritional quality that would, in turn, stimulate reproduction in biological control agent populations. KEY WORDS: Eichhornia crassipes; Neochetina eichhorniae; Neochetina bruchi; Phytophagy; Integrated control; Aquatic weeds PMID- 9852191 TI - ENVIRONMENTAL AUDITING: Assessing Biotic Integrity of Streams: Effects of Scale in Measuring the Influence of Land Use/Cover and Habitat Structure on Fish and Macroinvertebrates. AB - / Fish and macroinvertebrate assemblage composition, instream habitat features and surrounding land use were assessed in an agriculturally developed watershed to relate overall biotic condition to patterns of land use and channel structure. Six 100-m reaches were sampled on each of three first-order warm-water tributaries of the River Raisin in southeastern Michigan. Comparisons among sites and tributaries showed considerable variability in fish assemblages measured with the index of biotic integrity, macroinvertebrate assemblages characterized with several diversity indexes, and both quantitative and qualitative measurements of instream habitat structure. Land use immediate to the tributaries predicted biotic condition better than regional land use, but was less important than local habitat variables in explaining the variability observed in fish and macroinvertebrate assemblages. Fish and macroinvertebrates appeared to respond differently to landscape configuration and habitat variables as well. Fish showed a stronger relationship to flow variability and immediate land use, while macroinvertebrates correlated most strongly with dominant substrate. Although significant, the relationships between instream habitat variables and immediate land use explained only a modest amount of the variability observed. A prior study of this watershed ascribed greater predictive power to land use. In comparison to our study design, this study covered a larger area, providing greater contrast among subcatchments. Differences in outcomes suggests that the scale of investigation influences the strength of predictive variables. Thus, we concluded that the importance of local habitat conditions is best revealed by comparisons at the within-subcatchment scale. KEY WORDS: Stream; Biomonitoring; Land use; Scale; Habitat; Fish; Macroinvertebrates PMID- 9852192 TI - Modified GDP Through Health Cost Analysis of Air Pollution: The Case of Turkey. AB - / Economic growth and performance is monitored through the gross domestic product (GDP) of a nation. It has long been recognized that the traditionally computed GDP does not account for gains and losses observed due to the consumption of natural resources and environmental services. Hence, the objective of this study is to modify the Turkish GDP by taking social cost associated with air pollution into consideration. To this end health benefits and economic costs of air-quality improvement are estimated. The computations are based on the dose-response coefficients reported in several studies. The results indicate that a decrease in PM10 and SO2 levels to the WHO guideline would have resulted in a total of 48.309 x 10(10) and 153.38 x 10(10) Turkish lira savings in 1990 and 1993, respectively. These correspond to 0.12% and 0.08% of 1990 and 1993 GDPs, respectively. KEY WORDS: Health costs; Air pollution; Modified GDP PMID- 9852193 TI - Autoimmune diseases as stem cell disorders: normal stem cell transplant for their treatment (Review). AB - Using various animal models for autoimmune diseases, we have found that autoimmune diseases are stem cell disorders. The transplantation of normal hemopoietic stem cells (HSCs) can be used to treat autoimmune diseases, whereas the transplantation of abnormal HSCs from autoimmune-prone mice to normal mice leads to the induction of autoimmune diseases in recipients. To elucidate the differences between normal and abnormal HSCs, we have established a new method for purifying pluripotent-HSCs (P-HSCs), and have compared the qualitative differences. Although normal P-HSCs cannot proliferate under major histocompatibility complex (MHC)-incompatible microenvironments (stromal cells), abnormal P-HSCs can proliferate under such conditions. In addition, the proliferation of abnormal P-HSCs is much faster than that of normal P-HSCs. These findings indicate that abnormal P-HSCs are more resilient than normal P-HSCs. Therefore, we propose that allogeneic bone marrow transplantation (BMT) (not autologous BMT) should be applied to the treatment of autoimmune diseases. human data on BMT in autoimmune diseases is reviewed, and the conditions essential for successful BMT, including tolerance induction, are discussed. PMID- 9852194 TI - The energy substrate switch during development of heart failure: gene regulatory mechanisms (Review). AB - During cardiac hypertrophy and in the failing heart, the chief myocardial energy substrate switches from fatty acids to glucose. In this review, we describe recent progress in the elucidation of the molecular regulatory events involved in the dramatic downregulation of the expression of fatty acid utilization enzymes during development of cardiac hypertrophy and failure. Much of this work has focused on the gene encoding medium-chain acyl-CoA dehydrogenase (MCAD), which catalyzes a pivotal step in the mitochondrial fatty acid -oxidation (FAO) cycle. In vivo ventricular pressure overload studies performed in mice transgenic for human MCAD promoter fragments linked to reporter genes have shown that transcription is markedly downregulated within seven days of pressure overload. The temporal pattern of this alteration in MCAD gene expression has also been characterized in a rat model of progressive pressure overload-induced left ventricular hypertrophy (LVH) and heart failure (HF) [SHHF/Mcc-facp (SHHF) rat]. MCAD mRNA levels are downregulated (>70%) during both the LVH and HF stages in the SHHF rats compared with controls. In contrast, the activity and immunodetectable levels of MCAD enzyme were not significantly reduced until the HF stage, indicating additional compensatory control at the translational or post translational levels in the hypertrophied but non-failing ventricle. FAO enzyme expression was also shown to be downregulated in human subjects with dilated cardiomyopathy compared to age-matched controls. Taken together, these results have identified a gene regulatory program that is involved in the alterations in myocardial energy substrate utilization in the failing heart. The temporal correlation of diminished enzyme expression with onset of heart failure suggests that this alteration in lipid metabolism may play a role in the pathogenesis of pressure-overload induced heart failure. This gene regulatory pathway should be a useful target for experimental studies aimed at the molecular pathogenesis of the transition from stable cardiac hypertrophy to overt heart failure. PMID- 9852196 TI - Cardiolipin: biosynthesis, remodeling and trafficking in the heart and mammalian cells (Review). AB - Cardiolipin is the principal polyglycerophospholipid found in the heart and most mammalian tissues. This phospholipid is the only phospholipid localized exclusively to the mitochondria of mammalian cells. Cardiolipin appears to be involved, either directly or indirectly, in the modulation of a number of cellular processes including the activation of mitochondrial enzymes and hence production of energy by oxidative phosphorylation. The regulatory properties which govern cardiolipin biosynthesis, its remodeling and trafficking are beginning to emerge. Studies in the isolated perfused rat heart and H9c2 cardiac myoblast cells have indicated that the rate-limiting step of cardiolipin biosynthesis, via the cytidine-5'-diphosphate-1,2-diacyl-sn-glycerol pathway, is the conversion of phosphatidic acid and cytidine-5'-triphosphate to cytidine-5' diphosphate-1,2-diacyl-sn-glycerol. The cellular level of cytidine-5' triphosphate appears to control the production of cardiolipin in H9c2 cells. The activities of the other enzymes of the cytidine-5'-diphosphate-1,2-diacyl-sn glycerol pathway of cardiolipin biosynthesis in the heart may be modulated by thyroid hormone and unsaturated fatty acids. In addition, extra-mitochondrial cytidine-5'-diphosphate-1,2-diacyl-sn-glycerol and phosphatidylglycerol may be utilized for cardiolipin biosynthesis in the heart and permeabilized cells. Cardiolipin may be readily hydrolyzed by phospholipases and may be remodeled by a deacylation-reacylation pathway. Studies with a Chinese hamster lung fibroblast cell line CCL16-B2 have indicated that the remodeling of cardiolipin is markedly altered in the mitochondria of these cells and that this alteration in remodeling may be one of the underlying mechanisms for the mutation in oxidative energy production in these cells. Host cell cardiolipin may be trafficked from the mitochondria to an intracellular bacterial parasite Chlamydia trachomatis. The purpose of this review is to briefly discuss some of the more recent findings in cardiolipin metabolism in the heart and mammalian cells and to provide insight into their possible implications in the regulation of some cellular functions in mammalian tissues and cells. PMID- 9852197 TI - The role of eosinophils in pulmonary fibrosis (Review). AB - Pulmonary fibrosis is commonly characterized by inflammation of the alveolar wall, leading to derangement of normal alveolar architecture, and interstitial as well as intra-alveolar fibrosis. The process involves cellular interactions via a complex cytokine network and heightened collagen gene expression with abnormal deposition in the lung. Recent studies have identified a myriad of cytokines with potential roles in pulmonary fibrosis. Based on in vivo antibody neutralization studies, important roles for tumor necrosis a (TNFa), macrophage inflammatory protein 1a (MIP-1a) and transforming growth factor (TGF), have been established. The recent demonstration that the eosinophil is a major source for several of these key pro-fibrogenic cytokines during the early stages of fibrosis, strongly suggest a role for the eosinophil in pulmonary fibrosis. In vitro, eosinophils can elaborate factors capable of stimulating fibroblast proliferation, and their presence in lungs undergoing many forms of pulmonary fibrosis has been well documented. Further support for a role for eosinophils in pulmonary fibrosis are suggested by clinical data showing a correlation between lung eosinophil count and a poor prognosis and decreased responsiveness to therapy. This review will focus on the recent findings, which suggest novel potential roles for the eosinophil in the pathogenesis of pulmonary fibrosis. PMID- 9852198 TI - In vivo electroporation: a powerful and convenient means of nonviral gene transfer to tissues of living animals (Review). AB - Although in vivo electroporation is currently an unfamiliar nonviral means of gene transfer, accounting for only about 1% of total studies related to in vivo gene transfer and gene therapy, it may be extensively used for experimental and therapeutic purposes in the near future. Like other nonviral methods, in vivo electroporation has a variety of advantages over viral vectors as: any types of cells and tissues in theory could become a target, handling is easy and quickly done within a matter of second, repeated administration of DNA is possible, no immunogenicity is expected, and there is no constraints on amounts and sizes of DNA to be used. Gene transfer efficiency of in vivo electroporation was found to be equivalent to or even superior to that of in vivo lipofection, gene gun and direct DNA injection methods. Although gene expression exerted is transient and foreign genes are likely to be present in an episomal form when transferred by in vivo electroporation, foreign gene products could be detected for more than 1 month depending on tissues and DNA constructs used. Gene expression generated by in vivo electroporation could be controlled to a certain extent in a tissue- or cell-specific manner, and be induced as intended. Perhaps better appraisal of in vivo electroporation as a nonviral gene transfer method should be brought forth in the future after more detailed analyses. PMID- 9852195 TI - Stress proteins and glycoproteins (Review). AB - Proteins represent both structural and functional elements of biological organisms, however, their structural and catalytic function is directly linked to the acquisition and maintenance of a complex three-dimensional conformation. A molecular machinery to accomplish protein folding and maintenance in vivo is provided by a variety of molecular chaperones that include both heat shock proteins (Hsps), glucose-regulated proteins (Grps), and a separate class of stress glycoproteins (S-Gps). Different chaperones associate to form functional complexes (chaperone) and work coordinately to accomplish specific functions during the folding of particular proteins. In this review, we will summarize recently acquired new insights into the complexities of chaperones, the current state of S-Gps and their interactions with Hsps, and of specific chaperones that appear to be designed for the folding of cellular glycoproteins. Finally, we discuss the physiological role of chaperones by examining their function in specific cellular processes, namely tumor/host interactions and diseases associated with aberrant prion protein folding. PMID- 9852199 TI - Three-dimensional conformation of the epitopes of human superoxide dismutase-2 recognized by antibody against HIV-1p17. AB - We have previously reported that antibodies against HIV-1p17 protein cross-react with human superoxide dismutase-2 (SOD-2). To identify and characterize the epitopes of human SOD-2 recognized by anti-HIV-1p17 antibody, we synthesized several peptide fragments of human SOD-2 and found that the antibody did not bind to the LQPALK hexapeptide which is contained in both HIV-1p17 and human SOD-2. Instead, this antibody bound to four peptides which contain amino acid sequences similar, but not identical, to the known epitopes of HIV-1p17. These peptides have two features in three-dimensional (3-D) conformation: two protruded carbohydrate side chains on the peptide chain or a concave structure in the molecule. It is suggested that the antibody does not strictly recognize the amino acid sequence itself, but may recognize these 3-D conformations. PMID- 9852200 TI - Multiparametric analysis for the enumeration of CD34+ cells from bone marrow and stimulated peripheral blood. AB - Determination of CD34+ cells was performed in bone marrow and G-CSF mobilised peripheral blood samples. We adopted three different protocols of analysis: the Milan/Mulhouse protocol, the ISHAGE guidelines for CD34+ cell determination and our own protocol based upon the use of PAINT-A-GATEPRO software analysis program. An excellent correlation was demonstrated between the three methods (r2 0.98); however the analysis of variance showed a statistically significant difference between the results generated with the three methods (P=0.001). The differences between the three procedures are discussed with a special focus on the value of CD34+dim cells and the role of CD45 in the setting of a double staining. We have in fact identified a minor subset (CD34+CD38+CD45-) which would go unrecognised based upon its CD45 negativity. PMID- 9852201 TI - Characterization of the nuclear localization signal in the DNA helicase involved in Werner's syndrome. AB - The nuclear localization signal (NLS) of the DNA helicase involved in Werner's syndrome (WS) was studied. Previously, we noted that the C-terminal region of WS helicase contains the NLS. In this study, we generated in HeLa cells various chimeric proteins consisting of the N-terminal tagged with an enhanced green fluorescent protein and the C-terminal fragments of the WS helicase that were truncated either from N- or C-termini, and we examined the ability of fragments to transfer the fusion proteins to the nucleoplasm by fluorescence microscopy. A small C-proximal region containing 34 amino acid residues (residues 1369-1402) was found to contain full nuclear migration activities. Subsequent amino acid substitution experiments showed that a sequence of three positively charged amino acids (Lys1371-Arg1372-Arg1373) in this region are particularly important. Similar sequence has previously been defined as the nuclear localization signal of SV-40 large T antigen that also acts as a viral DNA helicase. Conservation of this motif was found in the C-terminal regions of the other RecQ type DNA helicases, including murine WS helicase, yeast sgs1 and rqh+1 and human Bloom syndrome DNA helicases. PMID- 9852203 TI - A 5'-monophosphate form of bredinin selectively inhibits the activities of mammalian DNA polymerases in vitro. AB - Bredinin is an immunosuppressive drug which is used clinically in Japan. In this study, we investigated bredinin's molecular mode of action to clarify its immunosuppressive effects. We focused on the DNA polymerases in the somatic DNA synthesis which may be required in the process of lymphocyte differentiation. We found that bredinin-5'-monophosphate (breMP) could be a potent inhibitor of mammalian DNA polymerase alpha(pol.alpha) and (pol.beta) in vitro, although bredinin itself has no such effects. BreMP inhibited the pol. alpha activity at less than 7 micrograms/ml and the pol. activity at 7 micrograms/ml. Neither breMP nor bredinin influenced the activities of a plant DNA polymerase, prokaryotic DNA polymerases such as E. coli DNA polymerase I and Taq DNA polymerase, or DNA metabolic enzymes such as DNase I, indicating that breMP selectively suppressed the activities of the mammalian DNA polymerases. For pol., beta breMP acted by competing with both the substrate and template-primer. For pol. alpha, it acted by competing only with the substrate, and non-competitively with the template primer. The ribose of bredinin is quickly and quantitatively converted to its ribose-5'-phosphate form in vivo as soon as it is incorporated into cells. The action mode of bredinin and its use as an immunosuppressive drug are discussed based on these results. PMID- 9852202 TI - Drosophila antibacterial protein, cecropin A, differentially affects non bacterial organisms such as Leishmania in a manner different from other amphipathic peptides. AB - The effects of the antibacterial protein Drosophila cecropin A on developmental forms of Leishmania were compared with the effect of Hyalophora cecropin A in vitro. Both cecropins had a potent lytic activity on the promastigotes at concentrations not far from those occurring in vivo in the respective insect. Drosophila cecropin A had strong differential effects on the two maturation forms of Leishmania aethiopica at high concentrations: inhibiting intracellular amastigotes and stimulating extracellular promastigotes to take up thymidine. Hyalophora cecropin A also inhibited amastigotes by up to 50% at concentrations of 0.250 mg/ml, and inhibited promastigotes at high concentrations but had no enhancing effects at any of the concentrations tested. In contrast to the results with Leishmania, Drosophila cecropin A had no discernible effect on any developmental stage of P. falciparium and showed no lytic effects on haemocytes. The two enantiomers of a synthetic amphipathic peptide, D- and L-KALA, were also tested. D- and L-KALA had some in vitro antimalarial effects at 0.025 and 0.05 mg/ml respectively but both forms were haemolytic at 0.1 mg/ml. Potential uses of naturally occurring proteins and their derivatives in the control of insect born infections and topical use of cecropins against leishmaniasis are discussed. PMID- 9852204 TI - Reduction of platelet phospholipase C-delta1 activity in Alzheimer's disease associated with a specific apolipoprotein E genotype (epsilon3/epsilon3). AB - The epsilon4 allele of apolipoprotein E (apo E) is increased among patients with sporadic or familial Alzheimer's disease (AD). We examined platelet phospholipase C (PLC)-delta1 activity in AD patients either homozygous for apoepsilon3 or having at least one apo epsilon4 allele. We found that platelet PLC-delta1 activity is reduced from control levels in patients homozygous for apo epsilon3, but not changed in patients with an apo epilson4 allele. Reduced PLC-delta1 activity and apo epsilon3 may contribute to AD pathogenesis apart from the apo epsilon4 allele. PMID- 9852205 TI - Correlation of activated monocytes or B cells with T lymphocyte subsets in patients with Graves' disease. AB - We analyzed the phenotypic characteristics of PBMC from 34 patients with Graves' disease (GD) at different stages of the disease to explore the sequence of immunological events associated with it. In all cases their monocytes were in a state of activation and differentiation more advanced than those of a group of 23 healthy individuals. Strikingly, some patients had CD14++DR- immature monocytes, which were absent in healthy individuals. CD14+CD16+DRhigh monocytes were more abundant in patients. We found a positive correlation between the CD14++DR- monocyte and CD4+CD45RA- helper cells and a negative correlation between the same monocyte subset and CD4+CD45RA+ naive cells. CD14+/++DRlow monocytes directly correlated with this latter T4 subset and CD14+ CD16+DRhigh with CD4+CD45RO+ memory lymphocytes. There was also a positive correlation between memory T4 cells and the subset of activated B lymphocytes (CD19+CD5+) and suppressor T8 cells (CD8+CD11b+). T8 cytotoxic cells (CD8+CD11b-) positively correlated with T4 naive cells. The circulating levels of T3 and TSI (thyroid-stimulating immunoglobulin) directly correlated with a decrease in naive cells and an increase in T8 suppressors. The results suggest that the imbalance suppression/cytotoxicity in GD may be due to a reiterated presentation of autoantigens, or mimetic antigens, to T helpers by mature monocytes and activated B cells. PMID- 9852207 TI - A progressive postoperative increase in the serum level of soluble receptors for interleukin-2 is an indicator of a poor prognosis in patients with gastric cancer. AB - Activated T lymphocytes release a soluble form of IL-2R (SoIL-2R) into the bloodstream, which can be detected by CD25 monoclonal antibody. Perioperative changes of serum levels of SoIL-2R and the number of CD25-positive cells were monitored simultaneously to clarify the clinical implications of SoIL-2R in patients with gastric cancer (n=91). Preoperative levels of SoIL-2R were significantly higher than in normal controls and levels were a useful indicator of possible lymph node involvement. Postoperative levels of SoIL-2R increased independently of the number of CD25-positive cells. Patients with progressive postoperative increases in levels of SoIL-2R had both a significantly high frequency of postoperative relapse and a poor prognosis. Increased SoIL-2R may reduce the availability of IL-2 by binding to it. Postoperative progressive increases in SoIL-2R appear to be a good indicator for a poor prognosis in patients with gastric cancer. PMID- 9852206 TI - The storage of noradrenaline, neuropeptide Y and chromogranins in and stoichiometric release from large dense cored vesicles of the undifferentiated human neuroblastoma cell line SH-SY5Y. AB - Sucrose gradient centrifugation combined with electron microscopy revealed that undifferentiated SH-SY5Y cells contain predominantly one population of noradrenaline containing vesicles, i.e. large dense cored vesicles. These vesicles have been purified approximately twenty times using sucrose/D2O gradients. Electron microscopy of sucrose/D2O fractions confirms that large dense cored vesicles are enriched in the fractions containing predominantly dopamine- hydroxylase, chromogranin A, noradrenaline and neuropeptide Y. The membranes of these vesicles contain the typical large dense cored vesicle markers dopamine- hydroxylase, synaptotagmin, cytochrome b561 and rab 3. Stimulation of SH-SY5Y cells with carbachol and KCl shows that noradrenaline and neuropeptide Y are released in the same proportion as stored in the large dense cored vesicles. The immuno-blot pattern and intensity of chromogranin A and chromogranin B present in large dense cored vesicles and in the released material were definitely the same. This suggests that noradrenaline and the proteins/peptides are released in the same molar stoichiometry as they are stored in large dense cored vesicles. These data provide for the first time experimental evidence that the neuroblastoma cell line SH-SY5Y contains functionally active large dense cored vesicles similar to those of sympathetic neurons and indicate that this cell line is a suitable experimental cell model to study the exocytotic pathway of large dense cored vesicles. PMID- 9852208 TI - Experimental dermatophyte infection abated by povidone-iodine: assessment by computerized-assisted corneofungimetry. AB - The effect of antiseptics upon fungal growth inside human stratum corneum is poorly documented. Therefore the present study was undertaken to assess the anti dermatophyte activity of a diluted povidone-iodine wash solution using the corneofungimetry bioassay. When compared to a non-medicated cleansing product, regular applications of the antimicrobial for two weeks significantly abated the fungal invasiveness inside human stratum corneum. The present findings are most likely relevant for the clinical practice. PMID- 9852209 TI - Apoptosis-gene expression in hematopoietic system: normal and pathological conditions (Review). AB - Gene expression involving apoptosis in the hematopoietic system is reviewed. In normal and hematological disorders, Fas-Fas ligand and tumor necrosis factor alpha-receptor interaction play a major role in enhancing apoptosis. On the other hand, bcl-2 or certain novel proteins (including FADD, RIP, TRADD and sentrin) prevent apoptosis. Apoptosis is involved in myelodysplastic syndrome and pathogenesis of leukemia. Expression of Fas antigen plays a role in negative regulation of hematopoiesis in the bone marrow as does interferon-gamma. PMID- 9852210 TI - Bcl-2- and CrmA-inhibitable dephosphorylation and cleavage of retinoblastoma protein during etoposide-induced apoptosis. AB - Cell numbers are regulated by a balance between proliferation and apoptosis (programmed cell death). Recent evidence suggests that proteins regulating cell proliferation also mediate apoptosis. Therefore, cellular fate might be determined by cross talk between regulators of cell cycle progression and apoptosis. Previously, we had found that during DNA damage-induced apoptosis, retinoblastoma protein (RB), an important G1/S regulator and tumor suppressor, became dephosphorylated and then immediately cleaved into p48 and p68 fragments. Here, we report that expression of the Bcl-2 oncoprotein, an inhibitor of caspases (interleukin 1 -converting enzyme-like proteases), blocked RB dephosphorylation, RB cleavage and apoptosis in etoposide-treated human Jurkat T cells. In addition, expression of the cowpox virus CrmA protein, a direct inhibitor of caspases, also inhibited both RB changes and apoptosis. Taken together, our findings demonstrate important roles for caspases in the processes of etoposide-induced RB dephosphorylation, RB proteolysis and apoptosis. PMID- 9852211 TI - The control of lytic replication of Epstein-Barr virus in B lymphocytes (Review). AB - Uncontrolled replication of a virus, which is harmful to the host is also disadvantageous to the virus. Most viruses cannot compete with the various immune mechanisms and become eliminated in the course of infection. Therefore, only the time between infection and eradication remains for these viruses to proliferate. A few viruses, like the Herpesviruses or the papillomaviruses, however, have developed a sophisticated strategy for persisting lifelong, usually asymptomatically in the host, hiding from the immune system and producing infectious progeny at the same time. This strategy depends on a separation of latency and the lytic replication, either by time due to differentiation dependent mechanisms or by spatial separation as the result of different host cell types. Both are true for the Epstein-Barr virus (EBV). B cells and epithelial cells have a pivotal role in the life cycle of the virus. The former can become latently infected and are thought to be the virus reservoir in vivo, whereas the latter were shown to be permissive for lytic replication. However, replication of EBV in vivo is controlled primarily by host immune mechanisms selecting for cells that are not permissive for viral replication as the result of a particular set of transcription factors. These factors control the activity of the regulatory immediate-early genes and, in addition, lytic and latent cycle regulatory genes negatively interfere with each other and thus link cellular and viral gene regulatory mechanisms. Disturbance of both the immune surveillance as well as viral gene regulation may result in EBV-associated disease. PMID- 9852212 TI - Dehydroepiandrosterone sulfate deficiency in chronic fatigue syndrome. AB - The chronic fatigue syndrome (CFS) is a condition of unknown etiology, characterized by a persistent debilitating fatigue, the muscle-related symptoms and the neuropsychiatric symptoms. Recently, it has been reported that the patients with CFS might have impaired activation of the hypothalamic-pituitary adrenal axis, and suggested that a part of the patho-genesis of CFS might be associated with abnormalities of the endocrine system. Herein, we show that the majority of Japanese patients with CFS had a serum dehydroepiandrosterone sulfate (DHEA-S) deficiency. Serum DHEA-S is one of the most abundantly produced hormones which is secreted from the adrenal glands, and its physiological function is thought to be a precursor of sex steroids. DHEA-S has recently been shown to have physiological properties, such as neurosteroids, which are associated with such psychophysiological phenomena as memory, stress, anxiety, sleep and depression. Therefore, the deficiency of DHEA-S might be related to the neuropsychiatric symptoms in patients with CFS. PMID- 9852213 TI - Stalling of DNA methyltransferase in chromosome stability and chromosome remodelling (Review). AB - As a consequence of their mechanism of action, DNA (cytosine-5) methyltransferases from both prokaryotes and eukaryotes necessarily recognize mispaired bases in unusual DNA structures as catalytic transition-state analogs. A review of the available data suggests that the enzymes are designed to stall at these sites because they are unable to release substrates or products that are fixed in a conformation resembling the transition state. The enzymes can operate by a two-step process in which they first methylate extrahelical cytosines satisfying their recognition requirements and subsequently stall at the site of methylation. On RNA and DNA RNA hybrids they may operate by a similar one-step process in which they stall at transition-state analogs without methylating cytosine moieties. These natural capacities suggest that the enzymes may physically participate in stable nucleoprotein assemblies formed as components of normal chromatin structure or as intermediates in the repair of unusual structures. The methyltransferases, themselves, may physically participate in chromosome remodelling as part of a mechanism of inactivation or imprinting by stabilizing RNA DNA hybrids or RNA RNA secondary structure involving cis-acting untranslated RNAs like the product of the Xist gene. Methyl-transferase may physically participate in the repair of certain unusual structures by serving as a nucleation point. The affinity for secondary structure in nucleic acids may account for the spreading of DNA methylation patterns. Titration of host methyltransferase by RNA DNA hybrids and RNA secondary structure formed during retroviral replication in certain tumorigenic retroviruses, like MMTV, may account for global hypomethylation observed in retrovirally transformed cells. In a similar fashion, titration of methyltransferase by secondary structures associated with chromosome instability may account for global hypomethylation observed in association with local hypermethylation in tumorigenesis. PMID- 9852214 TI - MHC class II-independent, Vbeta-specific activation of T cells by superantigen mutants fused to anti-tumor Fab fragments: implications for use in treatment of human colon carcinoma. AB - Genetically engineered fusion proteins of the super-antigen staphylococcal enterotoxin A (SEA) and tumor-reactive monoclonal antibodies, C215Fab-SEA and C242Fab-SEA, have been generated and shown to be effective in mediating superantigen-antibody directed cellular cytotoxicity against human carcinoma cells expressing the CA215 or CA242 antigens in an MHC class II-independent manner. In an attempt to reduce the in vivo toxicity of superantigen administration, alanine substitution mutations in SEA at residues F47 and D227 that affect SEA binding to class II molecules have been created and genetically linked to C215Fab or C242Fab. The purpose of this study was to determine whether these Fab-SEA mutant fusion proteins, that have low MHC class II binding affinities, were still able to stimulate human T cells in a Vbeta-specific manner in the presence or absence of MHC class II molecules. The SEA wt- and SEA-D227A based fusion proteins shared the ability to activate V beta5. 2-, Vbeta6-, Vbeta7 , Vbeta9- and Vbeta18-bearing T cells, whereas Fab-SEA-F47A protein activated only Vbeta6- and Vbeta7-bearing T cells. The fusion of Fab fragments onto SEA wt, SEA-F47A or SEA-D227A had no effect on the Vbeta specificity of these superantigens. Fab fusion proteins containing either SEA wt or SEA mutants were presented, in the absence of class II molecules, by CHO cells transfected with CA215 and CD80 and all induced the expansion of only Vbeta6-, Vbeta7- and Vbeta 18-bearing T cells. Fab-SEA mutant fusion proteins may provide attenuated therapeutic agents that, while still able to specifically target high affinity T cells for MHC class II-independent local tumor killing, will not induce excessive systemic toxicity. PMID- 9852215 TI - The molecular regulation of protein breakdown following burn injury is different in fast- and slow-twitch skeletal muscle. AB - We compared the effect of burn injury on the energy-ubiquitin-dependent proteolytic pathway in the fast-twitch extensor digitorum longus (EDL) and the slow-twitch soleus muscle in rats. Rats were subjected to a 30% total body surface area full-thickness burn or sham procedure. At various time points after injury, total and myofibrillar protein breakdown rates were determined in incubated EDL and soleus muscles. The energy-dependent component of protein break down was determined by incubating muscles in energy-depleting medium. Messenger RNA levels for ubiquitin and RC3, a 20S proteasome subunit, were measured by Northern blot analysis. Burn injury resulted in an approximately 50% increase in total protein breakdown and a 3-4 fold increase in myofibrillar protein breakdown in EDL muscles, and this response reflected increased energy-dependent protein breakdown. In contrast, protein breakdown rates were not significantly influenced by the burn injury in soleus muscles. Ubiquitin mRNA levels were increased almost 10-fold in EDL and approximately 4.5-fold in soleus muscles following burn injury. Burn injury resulted in a 2-fold increase in RC3 mRNA in EDL with no significant changes noted in soleus muscles. The results suggest that the more pronounced effect of burn injury on protein breakdown in fast-twitch than in slow twitch muscle may reflect different regulation of proteolysis at the molecular level. PMID- 9852216 TI - Interactions between human embryonic forebrain cells and the cytokines interferon gamma and interleukin-4. AB - In the present study we investigated the capacity of human first trimester forebrain cells at different gestational ages to produce IFN-gamma and IL-4. We also studied the effects of IFN-gamma on their proliferation, survival and expression of MHC antigens. IFN-gamma but not IL-4 was spontaneously produced after 24 h cultures. Furthermore, IFN-gamma exhibited anti-proliferative effects and induced MHC expression on these cells. However, the IFN-gamma exposed cultures showed significantly higher cell survival compared to un-exposed cultures. Co-culture with IL-4 blocked the IFN-gamma production and reversed its anti-proliferative effects. These interactions suggest important roles for cytokines in the survival, proliferation and differentiation of human embryonic and fetal forebrain cells. PMID- 9852217 TI - Cytokines and anti-cytokine autoantibodies during experimental african trypanosomiasis in mice with disrupted interferon-gamma and interferon-gamma receptor genes. AB - We studied cytokines and anti-cytokine autoantibodies (Aabs) during T.b.brucei infections in IFN-gamma-/-, IFN-gammaR-/- and wild-type mice. Increased serum levels of IFN-gamma, TNF-gamma and IL-4 with decreased Aabs to these cytokines were recorded early during infections in all mice (except IFN-gamma in IFN-gamma /- mice). Later, these responses were reversed, and surprisingly Aabs reacting to IFN-gamma in the IFN-gamma -/- mice were detected. To examine the possibility that an IFN-c immunoreactive molecule might be expressed due to infections and upon gene deletion, anti-IFN-gamma antibody was inoculated and resulted in abrogation of such Aabs. The scenario was different for IL-10 and TGF- since IFN gammaR-/- and wild-type mice showed low cytokines and high Aabs early during infections, but later high cytokines and low Aabs were registered. Interestingly, IFN-gamma-/- mice exhibited reversed levels of both IL-10 and TGF-beta, and also of their Aabs. Fab fragments of purified serum immunoglobulins showed binding and neutralizing effects in biological assays. Pre-absorption of the Fab fragments with a cytokine inhibited the binding and neutralization effects of this cytokine, but not of other cytokines. These results highlight an important role for autoimmunity in cytokine regulation, and that genomic deletion of IFN-gamma modulates cytokines and their Aab responses in experimental African trypanosomiasis. PMID- 9852218 TI - Immunoreactivity of sera to a peptide derived from the serotonin 5-HT1A receptor in a group of children with developmental disorders: possible role in non autistic epilepsy. AB - The presence of autoantibodies against the serotoninergic 5-HT1A receptor has been reported in serum from an autistic child using radioligand binding studies. It is now well established that, in cardiovascular diseases with an autoimmune component, patients present in their sera autoantibodies directed against the second extracellular loop of some G-protein coupled membrane receptors. We thus investigated by an enzyme-immunoassay method the presence of anti-5-HT1A receptor antibodies in sera of children with developmental disorders using synthetic peptides corresponding to the first and the second extracellular loops of this receptor. The population of children with developmental disorders was divided in autistic children with or without EEG abnormalities, and in non-autistic children with or without EEG abnormalities. We found that 6 out of 10 sera of non-autistic children with an abnormal EEG recognized the second extracellular loop of the 5 HT1A receptor. This is significantly higher than the other groups of children with developmental disorders or a healthy control group. These observations support the existence of an autoimmune component in epilepsy. PMID- 9852219 TI - CpG transition strand asymmetry and hitch-hiking mutations as measures of tumorigenic selection in shaping the p53 mutation spectrum. AB - By the genetic code, the average protein perturbation expected from a CpG-->TpG transition is strand-specific and smallest when it originates with the C on the transcribed (noncoding) strand. To distinguish the effects of selection from mutagenesis, we measured strand asymmetry for CpG-->TpG transitions fixed in active p53 genes and pseudogenes during vertebrate evolution, and for p53 genes from human tumors with one (singlet) and two (doublet) p53 point mutations. Mutagenesis appears to generate the transitions symmetrically while selection usually acts asymmetrically being most sensitive to the larger protein perturbations. Tumorigenic selection acting on the central domain of the p53 gene appears exceptional in that it often senses gain of function amino acid substitutions whose altered function is unrelated to degree of protein perturbation. In doublets, the selection on some gain of function substitutions is relaxed as evidenced by a return to the transition strand symmetry. PMID- 9852220 TI - Hypersensitive cytokine response to beta-amyloid 25-35 in astroglial cells from IL-1 receptor type I-deficient mice. AB - betaA25-35, a neurotoxic fragment of the Alzheimer beta-amyloid peptide (betaA), acts as a strong inducer of pro-inflammatory cytokines, such as interleukin (IL) 1 and IL-6, in glial cells. Since IL-1 is known to induce expression of both IL-1 and IL-6, we have investigated to what extent the induction of IL-1alpha and IL-6 by betaA25-35, is dependent on the IL-1 receptor type I (IL-1RI), the only known signalling IL-1 receptor. Primary astroglial cell cultures prepared from wild type and IL-1RI-deficient mice were incubated in the presence of betaA25-35 (100 microM) for 19 h, followed by analysis of mRNA levels of IL-1alpha and IL-6. Cell cultures treated with betaA25-35 showed a significant increase in mRNA levels for IL-1alpha and IL-6 and in addition increased levels of IL-1alpha immunoreactivity. A supersensitive IL-1alpha response was observed in astroglial cell cultures lacking the IL-1 RI as compared to betaA25-35 treated cell cultures from wild-type mice. In contrast the betaA25-35-induced increase of IL-6 was lower in the absence of IL-1RI. In conclusion, these results suggest that a functional IL-1 signal transduction is not necessary for induction of mRNA levels of IL-1alpha and IL-6 in astroglial cell cultures treated with betaA25-35, but that induction of IL-6 involves at least two distinct mechanisms, one of which occurs via activation of the IL-1RI. PMID- 9852221 TI - CD95 predicts responsiveness to tretinoin in acute promyelocytic leukemia. AB - We describe a predictive marker (CD95) for the responsiveness to tretinoin (RA) in acute promyelocytic leukemia (APL). Functional CD95 expression during RA treatment have been observed only in those patients who responded to RA. Expression of CD95 (Fas antigen), which plays a major role in apoptosis, was determined by fluorescence activated cell sorter (FACS) analysis. APL cases in which no enhancement of CD95 expression was observed showed no response to RA and did not obtain complete remission. We propose that CD95 can predict the clinical response to RA probably due to differentiation. PMID- 9852222 TI - CCAAT/enhancer binding proteins are critical components of the transcriptional regulation of hematopoiesis (Review). AB - The coordinated expression of four different CCAAT/enhancer binding proteins (C/EBPs), C/EBPalpha, C/EBPbeta, C/EBPdelta, and C/EBPepsilon constitutes a critical component of the myeloid differentiation program. C/EBPs are modular proteins, consisting of an activation domain, DNA binding domain and leucine zipper dimerization region. Recent studies including the analysis of mice deficient in several C/EBP proteins emphasize the effects of these molecules in hematopoiesis. C/EBPalpha is a master regulator of myeloid progenitors, C/EBPbeta plays an important role in macrophage and B-cell development, C/EBPgamma is involved in B-cell development, and C/EBPdelta is upregulated during myelopoiesis. Furthermore, C/EBPepsilon is a regulator of terminal differentiation of eosinophils and functional maturation of neutrophils. The formation of alternative combinations of tissue-specific and cell-stage specific C/EBP dimers may allow differential regulation of target genes in hematopoietic cells and commitment to distinctive hematopoietic lineages. PMID- 9852223 TI - Isolation and characterization of tubular basement membrane antigen common to humans and rats. AB - Using Brown Norway (BN) rats, we isolated and characterized the tubular basement membrane (TBM) antigens that are immunologically common to humans. The renal basement membrane (RBM) of BN rat, as an antigen source, was solubilized with 8 M urea instead of collagenase followed by extraction with 0.5 M NaCl. On frozen section-immunohistochemistry, the autoantibody obtained from BN rats, which had been immunized with human RBM and showed tubulointerstitial nephritis, bound to the TBM, the basement membrane of the Bowman's capsule, and the brush border of the proximal tubules, but not to the GBM of the normal BN rat kidney. Nephritogenic antigens were isolated by immunoaffinity chromatography using Sepharose-bound purified autoantibody. By Western blot analysis of the eluate, bands with molecular weight of 200 kDa and 180 kDa were positively reacted to anti-FX1A (brush border antigen) antibody and were apparently different from the major bands with molecular weight of 145 kDa and 130 kDa. The bands with molecular weight of 145 and 130 kDa showed major cross reactivity with antibodies to fibronectin and laminin. In contrast with these high molecular weight (HMW) bands, the major 60 kDa band with three minor bands showed no reactivity with any type of antibody tested. These results indicated that the non-enzymatic solubilization of RBM is one of the possible procedures for isolating the HMW form of antigens. These antigens may be epitopically modified pre-existing constitutions of the basement membrane and may play a role in the induction of tubulointerstitial nephritis. PMID- 9852224 TI - Altered binding of the NF-GMa transcription factor is involved in the upregulated production of TNF-alpha by macrophages of mammary tumor bearing mice. AB - Macrophages from D1-DMBA-3 mammary tumor bearing mice have profound alterations in various functions, i.e. diminished antigen presentation, decreased cytolytic activity and depressed synthesis of IL-12. In contrast, these cells display a significant enhancement in the levels of TNF-alpha, which may be involved in the cachexia observed in animals bearing large mammary tumors. The molecular mechanisms involved in the upregulation of TNF-alpha in macrophages from tumor bearers were investigated. The levels of TNF-alpha RNA were increased in macrophages from tumor bearers, but, this was not due to an increase in the RNA half-life. An analysis of the binding of transcription factors relevant to the TNF-alpha gene regulatory region by electrophoretic mobility shift assays (EMSA) revealed no differences in the binding of any NF-kB complex component between macrophages from normal and tumor bearing mice. Likewise, there were no changes in the binding patterns of SP-1 and NF-Y. In contrast, the binding of the transcription factor, NF-GMa, was altered in macrophages from tumor bearers. Our results and those reported in other models of disease suggest that the excessive production of cytokines in pathological conditions, could be the result of alterations of the production and/or binding of transcription factors. PMID- 9852225 TI - Identification of a neurotrophic sequence in erythropoietin. AB - Erythropoietin (Epo) is a hematopoietic factor that facilitates erythroid progenitor cell proliferation and differentiation. Recently, trophic effects of Epo have been observed in central cholinergic neurons. We have confirmed the neurotrophic factor activity of Epo and moreover, demonstrated sprouting and signaling by Epo in neural cells. Further, we have identified a 17-mer peptide sequence (epopeptide AB) in Epo (AEHCSLNENITVPDTKV) with activity similar to that of the holoprotein. This peptide induces differentiation and prevents cell death in both murine NS20Y and human SK-N-MC neuroblastoma cell lines. However, epopeptide AB does not promote the proliferation of erythropoietic cell lines or mouse primary spleen cells. The biological activities in neural cells were blocked by the addition of an antibody to the extracellular domain of the Epo receptor, indicating that the bioactive effects of epo-peptide AB in neural cells are Epo receptor mediated. Both epopeptide AB and Epo stimulated phosphorylation of ERKs in PC12 cells. When epopeptide AB or Epo was locally injected into mice, the frequency of motor end plate sprouting in adjacent muscles increased in a manner similar to that induced by CNTF. These findings indicate that neural cells and not hematological cells respond to a peptide sequence within erythropoietin and suggests that Epo may have separate domains for neurotrophic and hematotrophic function. PMID- 9852226 TI - Positron emission tomographic analysis of central dopamine D1 receptor binding in normal subjects treated with the atypical neuroleptic, SDZ MAR 327. AB - SDZ MAR 327 is a new neuroleptic agent with high in vitro affinity for dopamine D1 and D2 receptors. The goal of this study was to determine the effect of time after SDZ MAR 327 administration on central dopamine D1 receptor occupancy in healthy humans. Positron emission tomography (PET) with the dopamine D1 receptor ligand, [11C] SCH 23390, was performed in 6 male volunteers (age 22-34), in both the drug naive state and at 1, 2 and 4 h after a single oral dose of SDZ MAR 327 (9 mg). The pre and post drug treatment [11C] SCH 23390 dynamic data were analyzed using two different methods, each yielding a parameter proportional to the receptor density: i) a simple regional comparison approximating the specifically bound to free fraction, B/F; and ii) a two compartment, two parameter model yielding the apparent distribution volume DV". With both methods, a metabolite corrected arterial input function was used and the vascular fraction of tissue (Vb) was fixed at a previously determined value of 4%. Method I served as a qualitative comparison of the paired studies and demonstrated little difference between the pre and post drug conditions, method II also confirmed that there was no significant change in binding of [11C] SCH 23390 in the striatum. These data indicate that SDZ MAR 327 produces little if any effect on dopamine D1 receptor binding at the dose used. PMID- 9852227 TI - Construction and in vitro evaluation of RFT5(scFv)-ETA', a new recombinant single chain immunotoxin with specific cytotoxicity toward CD25+ Hodgkin-derived cell lines. AB - The data of a closed phase I/II trial in patients with resistant Hodgkin's lymphoma indicate promising results using a chemically linked anti-CD25 ricin-A immunotoxin (IT) (RFT5-SMPT-dgA). This IT is based on the high-affinity moab RFT5. Since recombinant DNA technology permits the readier production of large amounts of ITs, we constructed a new RFT5-based fusion toxin [RFT5(scFv)-ETA']. We isolated mRNA from the hybridoma cell line RFT5, synthesized first strand cDNA and performed RT-PCR. Amplified coding regions of the light and heavy chain variable domains were joined together with a synthetic (Gly4-Ser)3 linker. The resulting single chain variable fragment (scFv) was fused to a modified Pseudomonas aeruginosa exotoxin A (ETA') lacking its cell-binding domain I. After IPTG-induced expression in Escherichia coli, the 70 kDa His-tagged fusion protein [RFT5(scFv)-ETA'] was isolated by osmotic shock and sonication under denaturing conditions. The recombinant toxin was purified on a Ni2+-NTA chelating sepharose and eluted with 250 mM imidazole. Pooled protein was renatured, dialyzed and concentrated by precipitation. Binding properties of RFT5(scFv)-ETA' were assessed on the CD25-expressing cell line L540cy by ELISA, immunohistochemistry and FACS analysis. CD25-specific binding was confirmed by immunoprecipitation experiments with recombinant human IL-2 receptor alpha. The in vitro toxicity of the chimeric protein was tested on the Hodgkin-derived cell lines L540cy, L428, L1236, a monocyte cell line U937 and a Burkitt lymphoma cell line BL38. RFT5(scFv)-ETA' inhibited protein biosynthesis of L540cy and L428 cells by 50% at concentrations (IC50) of 18 and 12 ng/ml, respectively. CD25-specific toxicity was confirmed by competitive toxicity assays. These data confirm for the first time binding specificity and toxicity of a recombinant anti-CD25 immunotoxin, against Hodgkin-derived cell lines; its applicability on Hodgkin's lymphoma needs yet to be evaluated in vivo. PMID- 9852228 TI - Use of the bnx/hu xenograft model of human hematopoiesis to optimize methods for retroviral-mediated stem cell transduction (Review). AB - The potentiality of primitive human hematopoietic cells can be profoundly affected by in vitro culture. Due to the growing number of protocols proposed for stem cell gene therapy and ex vivo expansion, it is crucial to define methods to preserve the generative capacity of human stem cells in culture while promoting self-renewal divisions. Stem cell division, homing, and subsequent lineage development can only be studied definitively by marking of pluripotent cells, followed by tracking and clonal analysis of the progeny in a long-term transplantation system. We have developed a bnx/hu xenograft model, in which transduced human hematopoietic cells can be individually tracked into different lineages over the course of one year post-transplantation. The tracking is accomplished by single cell cloning of individual T lymphoid and myeloid progenitors recovered from the marrow of the mice, and clonal integration analysis by the sensitive technique of single-colony inverse PCR. All cells derived from a stem cell transduced by a retroviral vector will carry the unique restriction fragment length polymorphism (RFLP) created by the random integration event. We have used the bnx/hu xenograft system coupled with single-colony inverse PCR to determine that human stem cells require stromal support, fibronectin support with cytokines, or the presence of Flt3 ligand during a 72-h ex vivo culture to maintain the ability to sustain long-term multilineage hematopoiesis. PMID- 9852230 TI - Gap junctional communication and the tyrosine phosphorylation of connexin 43 in interaction between breast cancer and endothelial cells. AB - Abnormal gap junction communication (GJC) has been associated with carcinogenesis. We investigated the role of endothelial cell GJC and connexin 43 (Cx43), the main gap junction protein in these cells, during tumour cell extravasation. GJC was determined by the ability of cells to transfer Lucifer yellow, to neighbouring cells. Tumour-endothelial interaction was assessed by DiI assay. Connexin 43 expression and tyrosine phosphorylation were measured by immunoprecipitation and Western blotting. Co-culturing of ECV304 endothelial cells with human breast cancer cells resulted in a rapid and transient loss of communication competence of ECV304. This inhibition was maximal within 5 min. GJC was almost fully restored in 2 h. The co-culturing also resulted in an increase in the tyrosine phosphorylation of CX43. The pattern of phosphorylation was similar to the loss and the recovery of GJC in ECV304. We conclude that interaction of tumour cells with endothelium effectively inhibits GJC of endothelial cells, which is attributed to the increased tyrosine phosphorylation of connexin 43. This may contribute to the extravasation of tumour cells from the circulation, an essential step in the establishment of metastasis. PMID- 9852229 TI - The role of 12-lipoxygenase in pancreatic -cells (Review). AB - Leukocyte type 12-lipoxygenase (12-LO) catalyzes the conversion of arachidonic acid (AA; C20:4) to 12-hydroperoxyeicosatetraenoic acid (12-HPETE) and linoleic acid (LA; C18:2) to 13-hydroperoxyoctadecadienoic acid (13-HPODE). Previous studies have demonstrated that 12-LO, but not 5- or 15-lipoxygenase (5-LO, 15-LO respectively), is specifically expressed in pancreatic -cells and is involved in regulating glucose-stimulated insulin secretion. Lipoxygenase products also have been linked with inflammatory pathways in endothelial cells, kidney mesangial cells, inflammatory bowel disease, and corneal epithelial cells. Therefore, 12-LO may play a role in cytokine mediated inflammation in pancreatic beta-cells (i.e. beta -cell dysfunction and cytotoxicity). Cytokines such as IL-1 stimulate both de novo 12-LO protein synthesis and enzyme activity in pancreatic beta-cells. The products generated by 12-LO may ultimately be involved in cellular events that lead to lipid peroxidation. Hydroperoxide and free radical production in beta cells can activate intracellular signaling pathways that lead to cell death or may directly damage mitochondrial and plasma membranes. Increased 12-LO expression has also been found in islets from prediabetic Zucker fatty rats, a model that demonstrates insulin secretory defects similar to human type 2 diabetes. In this review, we present an overview of the 12-LO pathway in regulating glucose-stimulated insulin secretion in beta-cells as well as more recent data which supports the hypothesis that the 12-LO pathway participates in cytokine mediated beta-cell dysfunction and cytotoxicity. PMID- 9852231 TI - Cellular and molecular mechanisms in transplant arteriosclerosis (Review). AB - Although significant progress has been made towards the understanding of cellular and molecular mechanisms underlying the pathogenetic pathways of transplant arteriosclerosis, its knowledge is still not comprehensive. Nevertheless, experimental and clinical studies have enabled us to discover some of the complex processes involved in the progression and evolution of transplant arteriosclerosis. Despite the advances in transplantation immunology and atherosclerosis research, transplant arteriosclerosis still remains a major cause of allograft failure. A curative treatment, in order to inhibit or at least modify the development of transplant arteriosclerosis, is urgently needed. This review article highlights some of the more recent aspects of cellular and molecular pathology of transplant arteriosclerosis that may add to our current and future diagnostic and curative interventions. PMID- 9852232 TI - The safety of treatment with recombinant human erythropoietin in clinical use: a review of controlled studies. AB - Recombinant human erythropoietin (rhEPO) has now been approved for the treatment of renal anemia, anemia of prematurity, cancer-associated anemia, AIDS-associated anemia and as concomitant treatment for patients with or without autologous blood donation awaiting elective surgery. The purpose of this review is to provide an overview, based on the results of controlled studies, of the anticipated safety profile of rhEPO in various indications and to assess whether treatment with rhEPO influences the incidences of certain adverse events in these indications. The anticipated adverse events differ from indication to indication and generally reflect the corresponding underlying illness. With most indications, no relevant differences in the incidences of adverse events are observed between rhEPO and placebo-control/patients. Only in the rhEPO therapy of renal anemia is an increased incidence of hypertensive events observed in the rhEPO groups, a finding that is not reproduced with the other indications. The controlled studies forming the basis of this review provide no evidence of a relevant increase in the risk of thromboembolic events during rhEPO therapy. Overall, it may be stated that rhEPO treatment, where strictly indicated, is a safe form of therapy. As with any other treatment, the risk of side effects in certain predisposed patients must also be weighed against the desired clinical benefits. PMID- 9852233 TI - Mechanisms of immune cell-mediated tissue injury in inflammatory bowel disease (Review). AB - This review discusses the mechanisms and pathways of immune cell-mediated intestinal inflammation and tissue injury in inflammatory bowel disease (IBD). Our lack of understanding of how the mucosal immune system normally functions to maintain the balance between tolerance and immunity to innumerable dietary and bacterial constituents of the gut is perhaps the biggest obstacle to understanding the cause(s) of IBD, and to developing more effective treatments for these debilitating disorders. Evidence that abnormalities or disruptions in the interaction of immune cells and gut bacteria can trigger or contribute to changes in the composition, regulation and activity of the mucosal immune system that result in inflammatory immune responses and tissue injury are discussed. Based upon these studies, we propose a model to explain how a breakdown in regulation and failure to resolve immune responses in the gut mucosa results in persistent activation of T lymphocytes and other immune cells and the uncontrolled production of soluble inflammatory mediators that directly or indirectly produce the pathophysiological changes and tissue injury characteristic of IBD. PMID- 9852234 TI - Mosaicism for Charcot-Marie-Tooth disease type 1A: onset in childhood suggests somatic reversion in early developmental stages. AB - A 1.5 Mb duplication on chromosome 17p11.2 is typical for the great majority of patients suffering from Charcot-Marie-Tooth type 1A (CMT1A) disease. A female child of 4 years with clinical signs and symptoms of a demyelinating neuropathy was examined for the presence of this duplication. Analysis of MspI polymorphisms in DNA extracted from peripheral blood failed due to homozygosity for probes pVAW409R3a and pEW401HE. Also, no EcoRI/SacI 3.2 kb junction fragment or dosage difference with probe pLR7.8, characteristic of the CMT1A duplication, was found. However, fluorescence in situ hybridization (FISH) analysis with the PMP22 specific probe c132G8 revealed in peripheral blood lymphocytes 60% of interphase nuclei with CMT1A duplication indicating the probability of mosaicism. In interphase nuclei extracted from nerve tissue the duplication was detectable in 88%, in muscle tissue in 72% of the analyzed nuclei. This suggests the presence of a somatic CMT1A duplication mosaicism that can only be reliably detected by FISH. The early onset and severity of the phenotype indicates that the hypothesized somatic reversion is probably fixed to early developmental stages. PMID- 9852235 TI - Glucocorticoid effects on extracellular matrix proteins and integrins in bovine trabecular meshwork cells in relation to glaucoma. AB - The trabecular meshwork (TM) is a specialized eye tissue essential for regulation of the aqueous humor outflow and control of the intraocular pressure. Disturbances of TM cells may lead to elevated intraocular pressure and glaucoma. This study assessed the dexamethasone effects on levels of extracellular matrix proteins and their integrin receptors in bovine TM cells. Instillation of glucocorticoids such as dexamethasone is known to result in ocular hypertension. The histologic changes induced resemble those seen in glaucoma. Examination of the effects of glucocorticoid therefore may provide insights into the pathogenesis of glaucoma. TM cells in either tissue culture or organ cultures were treated with 0 (control), 0.1, or 1 microM of dexamethasone for 72 h. Immunostaining, Western, Northern and dot blot analyses showed that dexamethasone caused an increase in levels of fibronectin and collagen type IV in tissue cultured TM cells. Increased focal contacts were also observed but the levels of laminin and collagen type I were unaffected. The dexamethasone effect was similarly demonstrated in organ cultures, with the exception that collagen type I also was enhanced. These results suggest that dexamethasone modulates extracellular matrices in the TM. Glucocorticoid may exert its effect through such a modulation in the development of steroid glaucoma. PMID- 9852236 TI - Stimulatory effects of substance P on CD34 positive cell proliferation and differentiation in vitro are mediated by the modulation of stromal cell function. AB - Substance P (SP) is a neuropeptide widely distributed in the nervous system. Extensive study has shown SP stimulates production of various cytokines by bone marrow stromal cells, although, the role of SP in hematopoietic phenomena is still unclear. Recently, we established a human cloned stromal cell line, HAS303, which can support hematopoietic stem cell proliferation and differentiation in vitro. We used this culture system to examine the effects of SP. Expression of the mRNAs of neurokinin (NK)-1R, NK-2R and NK-3R, specific SP receptors, on HAS303 cells was demonstrated by the RT-PCR. CD34+ cells isolated from bone marrow were co-cultivated with HAS303 cells in the presence and absence of SP and the total hematopoietic cells and progenitors were counted every 5 days. Introducing SP (10(-8) M) to the co-cultures significantly increased the number of total cells and progenitors compared with control cultures. SP showed no enhancing activity on CD34+ cells cultured alone. SP also stimulated IL-3 dependent colony formation of whole bone marrow MNCs in a soft agar culture system, but showed no such activity on isolated CD34+ cells in this system. These observations suggest that SP stimulated HAS303 cells, activated HAS303 cells, and stimulated the proliferation and differentiation of CD34+ cells. Treating HAS303 cells with SP increased the intracellular Ca2+ concentration and stimulated production of G-CSF, GM-CSF, SCF and IL-6, but not IL-1alpha, IL-1beta and TNF alpha, but did not enhance proliferation. All these findings suggest that SP mediates hematopoietic cell proliferation and differentiation in vitro by activating stromal cell function. PMID- 9852237 TI - Histamine-induced calcium mobilization in single cultured cells expressing histamine H1 receptors: a relationship between its sensitivity and the density of H1 receptors. AB - Stimulation of histamine H1 receptors initiates the hydrolysis of phosphatidylinositides and results in the production of inositol (1, 4,5) triphosphate and intracellular Ca2+ mobilization. Although the mechanism for signal transduction via the H1 recptor has been extensively investigated, little is known about the correlation between the sensitivity of histamine-induced Ca2+ mobilization and the density of H1 receptors in cultured cells. Cytosolic free Ca2+ concentration ([Ca2+]i) after stimulation by histamine was monitored in single CHO and rat C6-glioma cells stably expressed with H1 receptors and astrocytoma 1321N1 cells using the Ca2+-sensitive dye Indo-1 and dynamic single cell imaging techniques (ACAS 570 laser cytometer). Both of the H1 receptor expressed CHO cells and C6-glioma cells were over 10 times more sensitive to histamine than astrocytoma 1321N1 cells in which H1 receptors were naturally present. The density of H1 receptors in the transfected cells was also more than 10-fold that of 1321N1 cells. In addition, inhibition of intracellular Ca2+ ATPase by thapsigargin elicited an increase in [Ca2+]i in H1 receptor overexpessed cells and astrocytoma 1321N1 cells with similar sensitivity. These data suggest that the sensitivity of Ca2+ mobilization by histamine in these cells was correlatively augmented with the increase in the density of H1 receptors. PMID- 9852238 TI - The role of HIV-1 activated leukocyte adhesion mechanisms and matrix metalloproteinase secretion in AIDS pathogenesis (Review). AB - The following is a review of an emerging topic in the literature which has led to new hypotheses regarding the mechanisms of pathogenesis of the various tissue specific AIDS associated syndromes. The fundamental hypothesis in this review proposes that HIV-1 is able to increase lymphocyte and monocyte localization in tissues where released HIV-1 proteins cause local tissue damage leading to any one of the various AIDS associated syndromes. It is also hypothesized here that syndromes associated with other lymphotrophic viruses result from the ability of these viruses to direct leukocyte extravasation of blood vessel walls and to initiate tissue specific pathogenesis. Further, it is suggested here that new concepts and strategies for delivering gene therapy to specific tissues can be derived from our understanding of the mechanisms through which lymphotrophic viruses localize in specific tissues. HIV-1 infection of lymphocytes and monocytes leads to increased adhesion of these cells to vascular endothelium and extracellular matrix molecules. In addition, HIV-1 infection of various leukocytes leads to increased secretion of extracellular matrix degrading matrix metalloproteinases. Increases in leukocyte adhesion and matrix metalloproteinase secretion are associated with the normal mechanisms through which leukocytes localize in tissues during inflammation. The ability of HIV-1 to activate leukocyte adhesion and matrix metalloproteinase secretion suggests that HIV-1 has evolved a way to take advantage of leukocyte inflammatory mechanisms in order to exit the blood stream and gain access to body tissues. The ability of HIV-1 to use infected cells to localize in various tissues may lead to the establishment of HIV-1 reservoirs in tissues. Such viral reservoirs may cause the various tissue specific AIDS associated syndromes. AIDS patients have been found to have elevated adhesion molecules (integrins, and cell adhesion molecules or CAMs) on their peripheral blood lymphocytes (PBLs). While there is little clinical evidence that the tissue localization of HIV-1 infected leukocytes are the cause of the HIV-1 related syndromes, studies in vitro and with animal models have shown that the HIV-1 gene products Tat, Rev and gp120 are potent neurotoxins. It has also been shown that Tat can contribute to the growth of cells from Kaposi's sarcoma lesions. Further, HIV-1 infected cells have been shown to secrete cytotoxic levels of a variety of growth factors and small molecules. Thus, it is likely that the localization of HIV-1 infected cells in specific tissues could contribute to the HIV-1 associated syndromes such as AIDS dementia, HIV-1 related interstitial lung disease, HIV-1 associated nephropathy, the HIV-1 wasting syndrome and perhaps AIDS associated Kaposi's sarcoma and hyperproliferative skin disorders. This review will examine studies in the literature which demonstrate that HIV-1 infection increases leukocyte adhesion and matrix metalloproteinase secretion. Clinical reports of AIDS patient's leukocyte integrin levels will also be reviewed and evidence that tissue localized HIV-1 infected cells could contribute to a variety of HIV-1 associated syndromes will be presented. PMID- 9852239 TI - Airway epithelial cells as regulators of airway inflammation (Review). AB - Airway epithelial cells (AEC) are known to play an integral role in the airway defense mechanism via mucociliary system as well as mechanical barriers. Recent studies further indicate that AEC can produce and release biologically active compounds including lipid mediators, growth factors, endothelin and a variety of cytokines/chemokines important in the pathogenesis of airway disorders. Human bronchial epithelial cells were isolated from normal and diseased states, and purely cultured in hormonally defined, serum-free medium. Culture supernatants of AEC contained detectable amounts of cytokines such as IL-1, IL-6, IL-8, G-CSF and GM-CSF. Proinflammatory cytokines IL-1 and TNFalpha generally upregulated expression and release of these cytokines. Moreover, human bronchial epithelial cells from patients with airway diseases such as asthma showed increased levels of mRNA for the cytokines. AEC are considered to interact with immune and inflammatory cells by direct adhesion as well as by humoral factors including cytokines. For example, eosinophil adhesion to AEC may be an important signal for the activation and degranulation of eosinophils. AEC is also believed to take part in the airway mucosal immunity by interacting with lymphocytes. Finally, AEC may play a crucial role in the processes of airway remodelling found in chronic airway inflammatory diseases. These findings strongly suggest that AEC are actively involved as regulators of airway inflammatory responses playing an important role in the pathogenesis of airway disorders, and become a target for therapeutic intervention. PMID- 9852240 TI - A transgenic strategy for analyzing the regulatory regions of the human prostate specific antigen gene: potential applications for the treatment of prostate cancer (Review). AB - Prostate-specific antigen (PSA) has been used clinically as a marker for the diagnosis and staging of prostate cancer due to its specific expression in prostate epithelial cells. In addition to its medical importance, its complex hormonal and tissue-specific regulation makes it an attractive model to study gene regulation. Two approaches have been applied to the identification of regulatory regions which confer this specific expression pattern. In vitro analysis of the regulatory regions of the human PSA gene using promoter reporter constructs and tumor cell lines has revealed a number of the DNA sequences involved in the hormone-dependent expression of PSA. We have pursued an alternative in vivo approach using transgenic animal technology, which is the focus of this review. Using this second approach, a transgenic mouse was generated using a 14 kilobase (kb) region of the human PSA gene encompassing the coding region and intervening sequences as well as 6 kb of upstream sequence and 2 kb of downstream sequence. This genomic DNA clone confers a PSA expression pattern in mice which appears to be very similar if not identical to that of humans, allowing us to investigate tissue-specificity and developmental regulation of PSA expression. In addition, these mice, for which PSA is a self antigen, provide a model to test the feasibility and efficacy of PSA-directed immunotherapy for prostate cancer. The further identification of the PSA regulatory regions important for tissue-specificity may ultimately allow the design of new therapeutics for the treatment of prostate cancer. PMID- 9852241 TI - Gram-negative and gram-positive bacterial products induce differential cytokine profiles in the brain: analysis using an integrative molecular-behavioral in vivo model. AB - Bacterial-derived products [e.g., lipopolysaccharide (LPS) from Gram-negative and muramyl dipeptide (MDP) from Gram-positive bacteria] are proposed to play a pivotal role in the generation of neurological and neuroinflammatory/immunological responses during bacterial infections of the nervous system. LPS and MDP may act through cytokines; cytokine-neuropeptide interactions may also be involved. Here, we investigated cytokine and neuropeptide mRNA profiles in specific brain regions in response to the intracerebroventricular administration of LPS and MDP. IL-beta1 system components (ligand, signalling receptor, receptor accessory proteins, receptor antagonist), TNF-alpha, TGF-beta1, glycoprotein 130 (IL-6 receptor signal transducer), OB protein (leptin) receptor, neuropeptide Y, Y5 receptor, and pro-opiomelanocortin (opioid peptide precursor) mRNAs were analyzed. The same brain region sample was assayed for all components. LPS and MDP administration induced significantly different behavioral and molecular profiles. LPS was significantly more potent than MDP in inducing anorexia and in up-regulating pro-inflammatory cytokines (IL beta1 and TNF-alpha mRNAs in the cerebellum, hippocampus and hypothalamus; MDP was more potent in up-regulating anti-inflammatory cytokine (IL-1 receptor antagonist and TGF-beta1) mRNAs. LPS and MDP also modulated hypothalamic IL-1 receptor mRNA components, but did not affect any of the neuropeptide-related components examined. The results suggest that the magnitude of neurological manifestations induced by LPS and MDP may involve the ratio between stimulatory and inhibitory cytokines, and this ratio may have implications for the neuroinflammatory/neurotoxic events associated with bacterial infections of the central nervous system. PMID- 9852242 TI - Vascular cell-derived fibrinolytic regulators and atherothrombotic vascular disorders (Review). AB - Fibrinolysis is essential for maintaining the fluency of blood flow. Attenuated fibrinolytic activity has been frequently detected in coronary artery disease, peripheral vascular diseases, diabetes, hyperlipidaemia and obesity. The biologically active product of fibrinolytic system is plasmin. Generation of plasmin is regulated by plasminogen activators (PA) and their inhibitors (PAI). Vascular endothelial and smooth muscle cells synthesize tissue-type and urokinase type PA (tPA and uPA) and their major physiological inhibitor, PAI-1. The production of fibrinolytic regulators is modulated by a number of biological factors related to thrombosis and atherosclerosis, including coagulation factors, hormones, growth factors, inflammatory mediators and lipoproteins. Several anticoagulants, including heparin, hirudin and hirulog-1, affect the production of fibrinolytic regulators in vascular cells. Studies in knockout mice demonstrated that mice deficient in PA or plasminogen are susceptible to thrombosis during inflammation or injury. Overexpression of uPA or deficiency of PAI-1 promotes neointima and aneurysm formation, which is probably due to active remodelling of extracellular matrix in vascular wall caused by excess plasmin. Long-term effect of treatment with thrombolytic agents or in atheroscleronic cardiovascular diseases remains to be defined. Future studies on determination of the role of PA and PAI in vascular remodelling may help understand the mechanism for neointima formation and orient the prevention of restenosis following vascular procedures. PMID- 9852243 TI - Possible protective and pathogenic roles of gamma delta T lymphocytes in HIV infections (Review). AB - alpha beta and gamma delta T lymphocytes are largely responsible for the specificity of coordinated immune responses that are of crucial importance for protection from exogenous invaders and elimination of endogenous aberrations. One of the prominent distinguishing characteristics of primate gamma delta T lymphocytes is that most of their T cell receptors for antigen (gamma delta TCRs) are, unlike alpha beta TCRs, capable of recognizing nonpeptidic antigens in an MHC-unrestricted manner. Another interesting feature is that the gamma delta T cell subpopulation displays profound qualitative and quantitative changes in individuals with various infectious diseases. For example, the most frequent human peripheral blood gamma delta T cell subset expressing Vgamma9Vdelta2 TCRs is functionally disabled and numerically decreased in some HIV-infected persons. The nonresponsiveness of Vgamma9Vdelta2 T cells is accompanied by their decreased IFNc and TNFa production. The overall level of gamma T cell activation at different stages of HIV-infection may be clinically relevant. At an initial stage of HIV-infection, the extremely potent antiviral cytotoxic activities of Vgamma9Vdelta2 T cells may limit the viral spread. At later stages of disease, Vgamma9Vdelta2 T cell dysfunction may contribute to the loss of resistance to opportunistic pathogens (such as atypical mycobacteria) and neoplasms (e.g., lymphomas) frequently associated with HIV-infections. Also, it is possible that chronic stimulation of gamma delta T cells may result in immunopathology. In particular, the massive immunologic activation that appears to be the major contributing element of AIDS immunopathogenesis could be, at least in part, driven by gamma delta T cells overstimulated by repetitive exposures to HIV. PMID- 9852244 TI - Interferon-gamma receptor deficiency: relationship between genotype, environment, and phenotype (Review). AB - Interferon-gamma receptor ligand binding chain (IFNgammaR1) deficiency is an autosomal recessive inherited immune disorder. Mutations in the IFNgR1 gene are associated with severe infections due to mycobacteria. However, several aspects of the phenotype of IFNgammaR1-deficient children have recently been found to vary from case to case. This review thus discusses the respective roles of the genotype and of the environment in determining phenotypic variations among affected children. PMID- 9852245 TI - Melanomas, from the cell cycle point of view (Review). AB - Transformation of melanocytes to metastatic melanoma cells is characterized by unrestricted proliferation under growth-factor-deprived conditions, genetic loss of cyclin dependent kinase (CDK) inhibitors (CKI, e.g. p16INK4A), and aberrant production of autocrine growth factors (e.g. basic fibroblast growth factor). The latter induces increased expression of positive CDK regulators (e.g. cyclin D1) and reduced expression of additional CKIs (e.g. p27KIP1). Combined, these events lead to sustained CDK activity and hyperphosphorylation/inactivation of the retinoblastoma tumor suppressor protein (Rb). The persistent Rb phosphorylation causes the accumulation of E2F and the transcription of its target genes whose products promote cell cycle progression. PMID- 9852246 TI - Cytotoxic action of 2-deoxy-D-glucose tetraacetate upon human lymphocytes, fibroblasts and melanoma cells. AB - The tetraacetate ester of 2-deoxy-D-glucose was found to cause a concentration related (10 microM to 1.0 mM) inhibition of activated T cell proliferation and decrease in viability of both unstimulated and anti-CD3 monoclonal antibody stimulated peripheral blood mononuclear cells. Comparable findings were made in primary fibroblast cultures initiated from human skin explants. Likewise, 2-deoxy D-glucose tetraacetate exerted a time-related (4-72 h) and concentration-related (0.1-1.0 mM) cytotoxic action upon human melanoma cells of the Colo 38 line. In this cell line, multiparameter flow cytometric analysis of cells stained with annexin V and propidium iodide revealed a necrotic, rather than apoptotic, form of cell death after exposure to 2-deoxy-D-glucose tetraacetate. PMID- 9852247 TI - Host H-2 haplotype modulates the induction of host-versus-graft disease after the induction of neonatal tolerance to H-2 alloantigens. AB - Mice injected at birth with semiallogeneic spleen cells develop a host-versus graft disease (HVGD) characterized by the polyclonal activation of donor B cells by alloreactive host CD4+ T cells, the production of autoantibodies (autoAb) and the development of an inmmunocomplex-mediated glomerulonephritis. It has been demonstrated that the recognition of MHC class II, but not class I or non MHC, alloantigens triggers the development of the autoimmune syndrome (AIS). The finding of different expression patterns of Ia molecules in different mouse strains, and a closed restriction of some immune responses to particular H-2 haplotypes, prompted us to analyze whether variations in the expressed MHC class II molecules modify the HVGD. First, newborn BALB/c mice received spleen cells from F1 hybrid mice obtained by mating BALB/c mice with several mouse strains differing in the H-2 haplotype. Second, spleen cells from different F1 mice were neonatally injected in mice of both parental strains. All groups of BALB/c mice injected with different combinations of F1 mice showed an HVGD with a very similar serological course. However, in some instances, duration was different when comparing both parental strains injected with spleen cells from the mutual F1 hybrids. These results suggest that host MHC, but not donor MHC haplotype may modulate the AIS associated with the induction of neonatal tolerance. PMID- 9852248 TI - The role of anti-endothelial antibodies in the immunopathogenesis of transplant associated coronary artery disease (Review). AB - Transplant coronary artery disease (TxCAD) is manifest as a diffuse, concentric intimal proliferation which results in occlusion of the allograft vessel lumen, and is responsible for limiting the long-term success of cardiac transplantation. The recent discovery of high circulating levels of anti-endothelial antibodies (AEAs) in patients with TxCAD has resulted in increased clinical and experimental research interests in understanding their patho-physiological roles in TxCAD. Increasing evidence suggests that AEAs are cross-reactive towards an endothelial protein doublet of 56-58 kDa which has now been characterised and identified as the cytoskeletal protein vimentin. Despite this recent progress the immunopathogenesis of TxCAD remains unclear. In this review recent developments and mechanisms of the involvement of AEAs in the immunopathogenesis of TxCAD are discussed. PMID- 9852249 TI - The role of the soluble p53 antigen and its autoantibodies as markers for diagnosis of colon cancer: a comparative study. AB - The role of various serological tumor markers, p53 soluble antigen and its autoantibodies, was evaluated for the detection of colon cancer in humans. An HPLC technique was used to measure serum levels of both forms of p53 protein after their partial isolation on gel fiberglass affinity chromatography columns. Tumor-associated proteins (TAP) in the form of either antigens or autoantibodies were about 4% of the total protein isolated from the serum of colon cancer patients. The absolute amount of each of the two types of TAP was also similar: 14.69 2.88 and 18.29 3.85 mg ml-1, respectively. The amount of p53 autoantibodies was higher than those of p53 antigen, but the difference was not significant: 9.35 3.48 and 6. 19 3.87 mg/ml, respectively (p>0.05). A good correlation was found between the total amount of tumor-associated antigens (TAA) and the amount of p53 antigen (r=0.69), total amount of IgG and the amount of p53 autoantibodies (r=0.46), and between both forms of p53 protein (r=0.46). A high coefficient of regression was found between the total amount of TAA and the amount of p53 antigen (b=2.4). Relationships between Duke's stage in colon cancer and the serum levels of p53 protein were weak: 0.33 and -0.32 for p53 antigen and its autoantibodies, respectively. Serum determination of p53 autoantibodies has no advantage over the determination of p53 antigen. Both forms of p53 protein can be isolated with extremely high accuracy for the pathological diagnosis of cancer (87-93%). Specificity of the method was proved using of commercial p53 PAb OD1: the GFG columns with this antibody were able to isolate the same proteins which were isolated by GFG columns with anti-p53 IgG. Moreover, the isolation of p53 protein was performed with higher effectiveness using the GFG columns with entrapped anti-p53 IgG than those columns with commercial DO1 PAb. PMID- 9852250 TI - Soluble fractions of tumor necrosis factor-alpha, interleukin-6 and of their receptors in toxic epidermal necrolysis: a comparison with second-degree burns. AB - Drug-induced toxic epidermal necrolysis (TEN) is a rare bullous disease characterized by severe epidermal necrosis and sloughing. Soluble TNF-alpha(sTNF alpha), soluble IL-6 (sIL-6) and their reactive soluble receptors (sTNF-Rp55 or R1, sTNF-Rp75 or-R2, sIL-6R) were quantified in blister fluid and serum of 6 TEN patients and 13 cases of second-degree burn. The amounts of sTNF-alpha, sTNF-R1 and sTNF-R2 were significantly higher in TEN blisters than in burns reflecting the probable involvement of the TNF-alpha system in the specific pathomechanism of TEN. The ratio sTNF-alpha/sTNF-R2 was significantly lower in TEN blisters than in burns. The concentrations of sTNF-R2 in TEN blisters and serums were significantly greater than those of sTNF-R1. This suggests a potential important role for sTNF-R2 in TEN by enhancing the cytotoxic effect of TNF-alpha. In addition, both sTNF-R1 and sTNF-R2 were significantly more abundant in TEN blisters than in serums, indicating that the TNF-alpha processing was mainly a local event in the TEN skin. No significant difference could be established for sIL-6 and sIL-6R between TEN and burns. Although a role for IL-6 cannot be ruled out, its production has no specific characteristics in TEN compared to burns. PMID- 9852251 TI - Use of the recursion formula of the Gompertz function for the quantitation of PCR amplified templates. AB - One common drawback of the currently used procedures to quantitate the polymerase chain reaction (PCR) is that the statistical evaluation of the experimental data depends on many, not just trivial, model assumptions. In the present study we report on an improvement in this crucial step of the quantitative PCR. The experimental design underlying the introduced method is exactly the same as in the case of the so-called PCR. However, by applying growth curve analysis based on the recursion formula of the Gompertz function the kinetics of the accumulation of the amplicon are estimated conjointly from data spanning both the and phases of the reaction. We demonstrate the method by determining the relative number of templates (a 206 bp segment spanning the exon 3 of the X-chromosomal murine Hprt-gene) contained in known orders of dilutions of DNA isolated from the spleen of the C57BL/6J-mouse. [32P]-dATP incorporation was used in duplicate experiments to quantify the amplicons as a function of amplification cycles. Our results: i) indicate that the accumulation of the PCR product as a function of PCR cycles follows a sigmoidal pattern compatible with the Gompertz growth model (P<0.0000001); ii) directly support the thesis that the kinetical pattern of accumulation of amplicons of a given DNA fragment does not depend on the number of corresponding DNA templates provided to the reaction; iii) permit a simple direct evaluation of the parallelity in the course of the accumulation of amplicons from different template numbers as a function of amplification cycles, which is a silent preposition in the evaluation of the so-called PCR; iv) allow an easy quantitation of the relative number of provided templates. PMID- 9852252 TI - DNA mismatch repair genes and their association with colorectal cancer (Review). AB - Mismatch repair genes are involved in increasing the fidelity of replication by specific repair of DNA polymerase incorporation errors. In Escherichia coli, the best studied mismatch repair (MMR) pathway is the methyl-directed long patch repair system which is mediated by three gene products; MutS, MutL and MutH. These are conserved in higher eukaryotes. Mutations in human homologues of these proteins have been shown to be implicated in hereditary non-polyposis colorectal cancer (HNPCC). Alterations in the coding regions of MMR genes result in a mutator phenotype with marked instability of microsatellite sequences, indicative of a deficiency in DNA repair. PMID- 9852253 TI - Regulation of B cell apoptosis by Bcl-2 and Bcl-XL and its role in the development of autoimmune diseases (Review). AB - Cell death is a common event during B cell development. The demise of developing B cells is a regulated process that serves to select cell populations bearing functional receptors and to remove cells that are no longer needed or potentially autoreactive. Bcl-2 and Bcl-XL, two members of the bcl-2 gene family of programmed cell death regulators with anti-apoptotic activity, are expressed in a highly regulated pattern during B cell maturation. Overexpression of Bcl-2 in developing B cells of transgenic mice, in the presence of T cell dependent costimulatory signals, results in the generation of a modified B cell repertoire and in the production of pathogenic autoantibodies. While disregulation of programmed cell death in B cells may cause autoimmune manifestations in mice, the involvement of such alterations in the pathogenesis of autoimmune diseases in humans merits further investigation. PMID- 9852254 TI - Cytosolic Ca2+ signalling in inflammatory neutrophils: implications for rheumatoid arthritis (Review). AB - Recognition of the ways in which neutrophil behaviour is regulated may be crucial for a full understanding of their role in inflammation and in rheumatoid arthritis. Although it is well established that changes in cytosolic free Ca2+ play a central role in triggering neutrophil responses, only recently has evidence accumulated which points strongly to the existence of two distinct Ca2+ pathways in neutrophils. One pathway is mediated by conventional agonists, such as formylated peptides, IL-8, C5a and PAF, and the other by cross-linking and immobilisation of surface receptors, such as integrins, and the Fc receptors, CD32 and CD16. In this review, we give evidence for these two signalling pathways in neutrophils, highlighting the roles of two Ca2+ storage and release organelles, one centrally located and stationary, and the other peripheral and mobile. We point out the significance of these two routes of Ca2+ signalling for the correct sequence of neutrophil responses, and suggest that aberration of this sequence could result in pathogenic neutrophil activation. PMID- 9852255 TI - Anthracycline-derived chemotherapeutics in apoptosis and free radical cytotoxicity (Review). AB - Anthracycline-derivatives are frequently used chemotherapeutics in treatment of numerous human malignancies. Anthracyclines are known for their complex cytotoxic mechanism involving i) inhibition of enzymes such as topoisomerase II, RNA polymerase, cytochrome c oxidase and others; ii) intercalation into DNA; iii) chelation of iron and generation of reactive oxygen species (ROS); iv) induction of apoptosis. Here, mechanistic aspects for successful cytostasis and for side effects, e.g. cardiomyopathy, are discussed. We emphasize recent developments in anthracycline-mediated apoptosis and focus on a well known representative, doxorubicin (adriamycin, adriblastin). We reflect on the role of oxidative stress and interactions with intracellular signaling pathways. PMID- 9852256 TI - Autosomal dominant Charcot-Marie-Tooth disease type 1A and hereditary neuropathy with liability to pressure palsies: detection of the recombination in Slovene patients and exclusion of the potentially recessive Thr118Met PMP22 point mutation. AB - Charcot-Marie-Tooth disease type 1 (CMT1) and hereditary neuropathy with liability to pressure palsies (HNPP) are the most frequent autosomal dominantly inherited disorders of the peripheral nervous system. The recessive inheritance is observed only exceptionally. Unequal crossing-over of misaligned flanking CMT1A-REP elements on chromosome 17p11.2 is the most frequent cause of the CMT1A duplication and of the reciprocal deletion in HNPP patients. Recently a recombination was noted. In our study 71 Slovene CMT1 patients from 36 families, 12 HNPP patients from 6 families and their 31 healthy relatives were analysed for the presence of these recombination mutations. In 29 of 36 unrelated CMT1 (81%) and in all 6 unrelated HNPP patients the duplication or the deletion, on chromosome 17p11.2-12 was detected. In 26 out of 29 duplication patients (CMT1A) (90%) a 3.2 kb EcoRI/SacI duplication junction fragment was observed. The analogous 7.8 kb EcoRI/EcoRI deletion junction fragment was found in 4 out of 6 unrelated HNPP deletion patients (67%). Overall we found a recombination mutation inside the in 86% of unrelated Slovene CMT1A and HNPP patients. One hundred and thirty-six DNA samples of the CMT1 and HNPP patients and of the healthy controls were negative for the potentially recessive Thr118Met PMP22 amino acid substitution. Dominantly inherited CMT1A duplications and HNPP deletions on chromosome 17p11.2 are thus, as in most other European countries, the most common mutations in Slovene CMT1 and HNPP patients. No signs of polymorphism or of potentially recessive mutation were found at the specific Thr118Met PMP22 site. PMID- 9852257 TI - Molecular dissection of B cell antigen receptor signaling (review). AB - In B lymphocytes, signaling through the B cell antigen receptor (BCR) contributes to cell fate decisions with different extents of receptor engagement leading to such outcomes as cell death, survival, or proliferation. During the past several years we have seen significant strides in our understanding of the signaling pathways that connect the BCR to the nucleus. Stimulation of the BCR leads to the activation of three types of intracellular protein tyrosine kinases Lyn, Syk, and Btk. Concerted action of these tyrosine kinases leads to the phosphorylation of multiple substrates and to activation of a variety of signaling pathways including phospholipase C-gamma, Ras, and phosphatidylinositol 3-kinase activation. The ability of B lymphocytes to react appropriately to a wide variety of environment stimuli requires a high degree of regulation on these multiple signaling pathways. PMID- 9852258 TI - A mutation in the lipoprotein lipase gene associated with hyperlipoproteinemia type I in mink: studies on lipid and lipase levels in heterozygotes. AB - Severe hypertriglyceridemia was previously observed in mink. Affected animals had no detectable lipoprotein lipase activity, but normal amounts of lipoprotein lipase protein in post-heparin plasma. We have now cloned cDNA for lipoprotein lipase from normal mink and identified a single point mutation in the affected animals which most likely explains the deficiency of active lipase. The mutation is located in exon 6 and results in a Pro214Leu substitution. In heterozygote mink the levels of lipoprotein lipase activity and mass in post-heparin plasma were lower than in normal mink, but could not be used to identify carriers of the mutation. In some tissues (heart, muscle, kidney and lung), lipoprotein lipase activity was decreased to about 50%. In adipose tissue there seemed to be a mechanism to compensate for the mutation, resulting in increased mass and approximately the same activity of lipoprotein lipase as in animals not carrying the mutation. Mink had high lipoprotein lipase activity and mass in kidneys, although the levels of mRNA in kidney were many fold lower than in adipose tissue. Mink had very low levels of cholesteryl ester transfer protein activity in plasma. This may contribute to the high levels of HDL in this animal species. PMID- 9852259 TI - The role of melatonin in the human fetus (review). AB - Melatonin, an indole amine, primarily derived from the pineal gland is secreted during the hours of darkness. Melatonin acts as a hormonal transduction of photoperiod influencing the timing of seasonal and daily (circadian) physiological rhythms. Maternal melatonin crosses the placenta and enters the fetal circulation providing photoperiodic information to the fetus influencing the subsequent circadian and seasonal rhythms of the offspring. The function of melatonin in humans is more obscure. However, melatonin has attained prominence as a treatment for disturbed circadian rhythms and sleep patterns which occur as a result of transmeridian travel, shift work or blindness. The biological clock, the hypothalamic suprachiasmatic nuclei (SCN), possesses melatonin receptors, in both the adult and fetal human. This concurs with the reported influence of melatonin on human circadian rhythmicity and indicates that this influence may begin in utero. Melatonin receptors are widespread in the human fetus and occur in both central and peripheral tissue from early in fetal development. Thus, the influence of melatonin on the developing human fetus may not be limited to entraining circadian rhythmicity. Considering the transplacental availability of melatonin to the fetus the ingestion of melatonin by pregnant women may be inadvisable. PMID- 9852261 TI - Structure of and signal transduction through interleukin-4 and interleukin-13 receptors (review). AB - We have recently demonstrated that two different forms of IL-4R exist; classical or alternative. The classical IL-4R is predominantly expressed in hematopoietic cells and consist of IL-4R and IL-2Rgammac (gammac) chains. On the other hand, alternative form of IL-4R is predominantly expressed in non-hematopoietic cells and consists of IL-4R and IL-13Ralpha' chains. Moreover, the alternative form of IL-4R is also utilized as a functional component IL-13R complex. It has been shown that the phosphorylation and activation of JAK3 tyrosine kinase is crucial for IL-4 activation of STAT6 in hematopoietic cells. However, we have recently demonstrated that non-hematopoietic cells lack JAK3 expression. We also demonstrated that in these cells, STAT6 activation is mediated through JAK1 and JAK2 tyrosine kinases instead. Furthermore, our results show that IL-4 and IL-13 signals are transmitted through the alternative form of IL-4R in these cells. Thus, major differences exist between hematopoietic and non-hematopoietic cells with regard to structure and signal transduction through IL-4R and IL-13R systems. PMID- 9852260 TI - Telomerase activity as a novel marker of lung cancer and immune-associated lung diseases. AB - Telomerase maintains telomere length and is considered to be necessary for the indefinite proliferation of human cells. Telomerase activity is detected not only in germline and immortal cancer cells, but also in stem/progenitor cells of renewal tissues and activated lymphocytes. While it is generally agreed that telomerase is a useful tumor marker, the utility of telomerase activity in non cancerous cells should also be considered. In the present study, we quantitatively examined telomerase activity in 56 cytology samples and 106 bronchoalveolar lavage samples obtained from patients with various respiratory diseases. Fourteen of 34 samples obtained from lung cancer patients showed detectable telomerase activity, while only 7 of 128 samples obtained from patients without lung cancer did (p<0.001). Moreover, 12 of 14 telomerase positive samples with lung cancer showed strong signals, while none without lung cancer did. Among 106 non-cancerous bronchoalveolar lavage samples, 4 telomerase positive samples had increased number of lymphocytes and increased disease progression. These findings indicate that evaluation of telomerase activity may not only be a useful diagnostic test for lung cancer, but may also be a marker of disease aggressiveness for immune-associated lung diseases. PMID- 9852263 TI - A heterozygous germline mutation of the PTEN/MMAC1 gene in a patient with Cowden disease. AB - Cowden disease, or multiple hamartoma syndrome, is an autosomal dominant inherited cancer syndrome with a high risk of thyroid and breast cancers. Its susceptibility gene has been mapped to chromosome 10q22-23. Because a newly found tumor suppressor gene, PTEN/MMAC1, often mutated in glioblastoma and in prostatic and breast cancers, has been mapped to the same chromosomal locus, it is suspected that it may be the gene responsible for Cowden disease. germline mutations of the gene have been reported in 4 of 5 families with Cowden disease. We performed a genetic analysis of the PTEN/MMAC1 gene in a sporadically found patient with the disease who had no apparent family history of the disease. We found a germline heterozygous mutation of the PTEN/MMAC1 gene in a patient with Cowden disease. The mutation, a C to T substitution of a single base at codon 130, leads to a formation of stop codon, generating a truncated protein lacking both protein phosphatase signature motif and tensin-like domain. Our finding supports the hypothesis of the PTEN/MMAC1 gene as being responsible for Cowden disease even in a sporadic case. PMID- 9852262 TI - Establishment of a new human megakaryoblastic cell line, CMY, with chromosome 17p abnormalities. AB - A new megakaryoblastic cell line CMY was established from a Down's syndrome patient suffering from acute megakaryoblastic leukemia. The karyotypes of CMY showed deletion of chromosome 17 or the translocation of 17p, whereas the blasts of the patient did not reveal these abnormalities of chromosome 17 by conventional karyotype analysis. Blasts of the patient failed to respond to chemotherapy and complete remission could not be attained. The abnormalities of 17p became progressively predominant in the patient. These results suggest that the blasts of a minor clone which had the abnormalities of chromosome 17p might have existed in the patient from the beginning and CMY was established from the minor clone. Investigation of p53 gene by PCR-SSCP analysis revealed that blasts of the patient showed normal patterns, while CMY showed an abnormally migrating band in exon 5 alone. This result suggests that another novel oncogenic factor(s) besides p53 might be present on chromosome 17p and other tumor suppresser genes need to be studied. PMID- 9852264 TI - Classical and molecular cytogenetic methods in diagnosis of a rare translocation t(3;21). AB - We report the characterization by traditional cytogenetic methods and fluorescence in situ hybridization (FISH) of a rare balanced reciprocal translocation t(3;21) in a male spouse connected with several miscarriages. G- and C-banding analysis of the male karyotype was suggestive of breakpoints in chromosomal bands 3q11.1 and 21p11.1. Multicolor-FISH analysis using chromosome specific alpha-satellite probes in combination with a locus-specific bacterial artificial chromosome (BAC) from 21q22.3 allowed us both to identify the origin of centromeres in the derivative chromosomes and to establish centromere-telomere orientation of the translocated chromosome 21 segment. Characterization of the translocated chromosomes by FISH analysis permitted describing the exact karyotype of the carrier as: 46,XY,-3,-21,+der(3)(21qterO21q11. 2::3p11.2O cenO3qter),+der(21)(3pterO3p11.2::21q11.2OcenO21pte r). PMID- 9852265 TI - Detection of cancer cells in the blood by FACS sorting of CD45- cells. AB - We describe a simple and sensitive method for detection of low number of cancer cells in the blood. The method is based on FACS sorting of leukocytes labelled with anti-CD45 monoclonal antibody and examining CD45- cells by conventional cytology and immunostaining for cytokeratin 18. In a model study, cancer cells seeded at the frequency of 1 per 106 and 1 per 107 leukocytes were detected in CD45- population. Sensitivity of this method was comparable to reverse transcription polymerase chain reaction (RT-PCR) used for detection of cancer cells expressing CD44 variants-mRNA. In a pilot study, cancer cells were also isolated from the blood of some patients with locally advanced gastric cancer. This method may be useful for detection of circulating tumour cells in cancer patients. PMID- 9852266 TI - Intestinal and hepatic cell kinetics and mucous changes in vitamin-A-deficient rats. AB - Changes in tissue histopathology, cell morphology and kinetics and the number of mucus secreting cells were evaluated in the duodenum and colon of normal and vitamin-A-deficient rats. A lower number of goblet cells was detected both in the colon and in the duodenum of vitamin-A-deficient rats in comparison with control rats (p<0.05). vitamin-A deficiency was found to increase the proliferate rate of mucous cells in the duodenum significantly. No significant difference was noted in the proliferate index in the colon specimens in either control group of experimental rats. We suggest that the higher number of proliferating cells in the duodenal crypt and villi in the vitamin-A-deficient animals indicates that vitamin-A deficiency affects the cell proliferation and therefore could be of some importance in understanding the potency of retinoids in differentiation and their association with formation of epithelial metaplasia. PMID- 9852267 TI - Pathologic characterization of hypotensive C57BL/6J-agt: angiotensinogen deficient C57BL/6J mice. AB - a fpreviously produced angiotensinogen-deficient mice, i.e. mice with deleted renin-angiotensin system (RAS), with a genetic background on C57BL/6J - C57BL/6J agt (-/-) -, but no C57BL/6J-agt (-/-) which survived long enough to be weaned. In the present study, we attempted to prevent neonatal death and analyzed pathological development in C57BL/6J-agt (-/-). We indicate that mortality in C57BL/6J-agt (-/-) derived from C57BL/6J-agt (+/-) can be reduced by hypodermic saline injection in the 7 days following birth, that hydronephrosis developed by day 14 in association with polydiplasia and polyuria by day 30, and that chronic hypotension occurs. Hydronephrosis is less damaging to electrolyte resorption in younger mice, but not in adults. We also observed that C57BL/6J-agt (-/-) derived from C57BL/6J-agt (-/-) frequently develop fetal hydronephrosis and die of respiratory failure at birth. These results suggest that maternal RAS is associated with structural maturation of kidney and lung in late fetus and that postnatal RAS plays important roles in structural and functional maintenance of the kidneys. PMID- 9852268 TI - Vertical transmission of human immunodeficiency virus type 1: frequency and correlation of transmission (review). AB - Child bearing age women constitute the fastest growing subset of the human immunodeficiency virus (HIV) infected population. Consistent with this fact is the increasing number of HIV infected children. Vertical transmission from mother to child is the most common route of infection. Because of the nature of the virus, it is difficult to determine the timing of infection. There are three time periods during which infection can occur: prenatally, perinatally, or postnatally. Evidence for each is presented within this review. PMID- 9852269 TI - Detection of differential gene expression in human osteoblastic cells by non radioactive RNA arbitrarily primed PCR. AB - The aim of the present study was to detect differentially expressed genes in the human osteoblast-like osteosarcoma cell line SaOS-2 using non-radioactive RNA fingerprinting (RNA arbitrarily primed polymerase chain reaction, RAP-PCR). RNA was isolated at different time points from SaOS-2 cells grown with and without dexamethasone (DEX). By RAP-PCR we detected changes in band patterns of cells treated with DEX compared with untreated cells. PCR fragments further characterized and sequences from three of these gave perfect matches to the coding sequences of the human nucleophosmin gene B23, cDNA clone 4_c6 from P1 H25 and the human TRA1 gene, respectively. differential regulation of these genes in DEX-stimulated SaOS-2 cells could be demonstrated by RT-PCR. PMID- 9852270 TI - A novel autoantibody in paraneoplastic sensory-dominant neuropathy reacts with brain-type creatine kinase. AB - We have previously reported on a novel autoantibody in a patient with paraneoplastic sensory-dominant neuropathy. This autoantibody immunostains the rat primary sensory system and reacts with a 47 kDa protein on immunoblotting. Here, we report on the isolation from rat spinal cord of a molecule that is recognized by this autoantibody. By ammonium sulfate cut and gel filtration, affinity and ion exchange chromatographies, the immunoreactive protein was purified to homogeneity and identified as brain-type creatine kinase (B-CK). Our study revealed that the autoantibody of the patient reacted with B-CK in the primary sensory system. PMID- 9852271 TI - AZT side effect on mitochondria does not depend on either inhibition of electron flow or mitochondrial uncoupling. AB - The mitochondrial myopathy associated with long-term AZT therapy limits the clinical efficacy of this drug in AIDS therapy. Thus, in order to determine how AZT can affect mitochondria bioenergetics, the capability of AZT to both uncouple oxidative phosphorylation and inhibit electron flow in isolated rat liver mitochondria was investigated. The failure of AZT to oxidize intramitochondrial pyridine nucleotides, to stimulate mitochondrial swelling in K+-acetate plus valinomycin or to cause ATP hydrolysis shows that AZT is not an uncoupler. PMID- 9852272 TI - Increased serum levels of tumor necrosis factor-alpha are correlated to soluble intercellular adhesion molecule-1 concentrations in non-small cell lung cancer patients. AB - Tumor necrosis factor (TNF)-alpha has been shown to induce shedding of ICAM-1. Experimental studies report that soluble intercellular adhesion molecule-1 (sICAM 1) may interfere with the host immunesurveillance system. Serum levels of TNF alpha and sICAM-1 were determined by ELISA in 112 non-small cell lung cancer (NSCLC) patients. Serum concentration of TNF-alpha and sICAM-1 were related to tumor burden and progression; a significant correlation was observed between circulating levels of TNF-alpha and sICAM-1. Our study suggests that ICAM-1 could be a marker of TNF-alpha activity and that high levels of these molecules may have a prognostic value in lung cancer. PMID- 9852273 TI - Enhancement of liposomal gene delivery in human breast cancer cells by dimethyl sulfoxide. AB - Non-toxic concentrations ( 1%) of dimethyl sulfoxide (DMSO) enhance the liposomal delivery of DNA to both MCF-7 and MDA-MB-231 human breast tumor cells. Uptake of SV-40-luciferase was enhanced in MCF-7 cells by 14-fold while uptake of CMV-beta galactosidase was increased 10-fold. In MDA-MB-231 cells, uptake of SV-40 luciferase was increased by approximately 70%. A mixture of ethanol and polyethylene glycol (45:55) at a concentration of 1% produced less pronounced improvements in transgene delivery to MCF-7 cells (a 70% increase in SV-40 luciferase uptake and a 4-fold increase in CMV-beta-galactosidase uptake) but no improvement in SV-40-luciferase gene delivery to MDA-MB-231 cells. These studies suggest that select pharmaceutical adjuvants which dissolve clinically useful drugs may have promise as non-toxic vehicles for improving transgene delivery. However, the relative effectiveness of these adjuvants is likely to vary depending on both the nature of the gene being delivered as well as the tumor cell which is the target for uptake of the exogenous gene. PMID- 9852274 TI - Analysis of the DNA content in Bowen's disease. AB - The aim of this study was to evaluate the DNA content in Bowen's disease in comparison to healthy epidermis applying image cytophotometry. The material investigated was derived from 50 patients with Bowen's disease and 10 patients with healthy skin. For comparison of both groups the Kruskal-Wallis test was applied. Slides were stained with Feulgen and were evaluated with CAS-200 image analyzer. Only 7/50 morbus Bowen cases represented euploid histogram. The others 43/50 were either conspicious to be aneuploid (29/50) or clearly aneuploid (14/50). In contrast, all normal epidermis (10/10) were clearly euploid. Morbus Bowen cases demonstrated significantly higher 5c exceeding rate (p=0.0012) and significantly more cells in the S-phase (p=0.017). High aneuploidy rate and increased proliferative activity in morbus Bowen cases support the classification of these lesion as carcinoma in situ. PMID- 9852275 TI - Alternative splicing of ERCC1 and cisplatin-DNA adduct repair in human tumor cell lines. AB - Alternative splicing is a common natural tool for the inhibition of function of full length gene products. We explored whether there was evidence that alternative splicing of ERCC1 may serve such a function for nucleotide excision repair. The ratio of alternatively spliced species to full length species was assessed for the protein and/or for the mRNA, for a series of human cell lines and tissues. This ratio was plotted against the amount of cisplatin-DNA adduct repair in each cell line (n=9), as measured by atomic absorbance spectrometry. As the percentage of alternatively spliced protein and/or mRNA increased, the amount of cisplatin-DNA adduct that was repaired was reduced. This inverse relationship was associated with a substantial amount of scatter (r=0.635), particularly at low levels of repair. These data demonstrate an association between alternative splicing of ERCC1, and reduction in cellular capability to repair cisplatin-DNA adduct. PMID- 9852276 TI - Regulation of cell adhesion, a central mechanism in the anticancer action of essential fatty acids (review). AB - Certain members of essential fatty acids (EFAs) are known to have anticancer functions. It has been established in recent years that some of these anticancer actions are via the effects of these fatty acids on the adhesive properties of cancer cells, including cell-cell and cell-matrix adhesions. These discoveries have implicated EFAs as a group of novel agents, able to regulate adhesive function of cancer cells and thus bearing clinical importance. PMID- 9852277 TI - Effects of islet hormones on nerve-mediated and acetylcholine-evoked secretory responses in the isolated pancreas of normal and diabetic rats. AB - This study employs the pancreas of normal and diabetic rats to investigate the relationship between the endocrine and exocrine pancreas in the control of exocrine secretion employing enzyme and immunohistochemical and physiological techniques. Acetylcholine esterase (ACh-E) positive nerves were distributed in the interacinar regions of the pancreas lying close to the exocrine cells. There was no difference between the cholinergic innervation of the pancreas in normal and diabetic rat. Insulin (INS) immunopositive cells were observed in the peripheral and central portions of the Islet of Langerhans in the pancreas of normal rat. In the diabetic animals the number of INS-positive cells were decreased. In contrast, glucagon (GLU) and somatostatin (SOM)-immunopositive cells were identified mainly in the peripheral parts of the Islets of Langerhans and their numbers increased markedly in the diabetic pancreas. Insulin alone had no significant effect on amylase secretion in the normal pancreas whereas GLU and SOM evoked small increases in amylase out compared to basal. In contrast, the islet hormones have no detectable secretory effect on the diabetic pancreas compared to control. Both electrical field stimulation (EFS) of intrinsic secretomotor nerves and exogenous application of acetylcholine (ACh) resulted in marked increases in amylase secretion. In pancreatic acini and acinar cells ACh evoked dose-dependent increases in amylase release. In normal pancreatic segments a combination of either INS or GLU with EFS or ACh resulted in marked potentiation of amylase output. In contrast, SOM inhibited the EFS-evoked amylase output but enhanced the secretory response to ACh. In pancreatic acini and acinar cells from normal rat and in pancreatic segments from diabetic rats, the islet hormones had no potentiating effect on the ACh-evoked secretory response. Similarly, in the diabetic rat the islet hormone had no effect on EFS-evoked amylase output. In fura-2 loaded pancreatic acinar cells ACh-induced a marked increase in intracellular free calcium concentration [Ca2+]i compared to basal. Either INS or GLU, but not SOM, elicited a small increase in [Ca2+]i. Combining either INS or GLU with ACh resulted in a potentiation of [Ca2+]i compared with ACh alone. In contrast, SOM had no significant effect on the ACh-induced [Ca2+]i compared to the response obtained with ACh alone. In pancreatic acinar cells of diabetic rat ACh-elicited similar magnitude of [Ca2+]i compared to acinar cells of normal rat. However, when the islet hormones were combined with ACh there was no enhancement of [Ca2+]i compared to ACh alone. The results indicate that the potentiation of either EFS or ACh-evoked secretory responses by the islet hormones seem to occur only in pancreatic segments which have intact viable Islets of Langerhans and not in either acini and acinar cells or from the pancreas of diabetic rat. Moreover, it is apparent that cellular Ca2+ is involved with the interaction of ACh with either INS or GLU. PMID- 9852278 TI - The human polyomavirus, JCV, and neurological diseases (review). AB - JC virus, a human neurotropic polyomavirus, is the established etiologic agent of the fatal demyelinating disease, progressive multifocal leukoencephalopathy, which usually affects individuals with defects in cell-mediated immunity. Cytolytic destruction of oligodendrocytes, the myelin-producing cells of the central nervous system is attributed as the mechanism by which JCV induces demyelination. PML was at one time a rare complication however it is now a much more common disease affecting patients of all ages due to the prevalence of AIDS. Of interest, in vitro evidence points to a cooperative interaction between JCV and HIV-1, via the HIV-1 regulatory protein, Tat. In addition, JCV has been demonstrated to induce tumors of neural origin in several animal models, including rodents and non-human primates, and several clinical reports have suggested a possible association between JCV and glial-origin tumors in humans. The viral regulatory protein, T-antigen, which has been shown to play a key role in orchestrating the events of the viral lytic cycle, is also capable of altering cellular functions, by nature of its direct interaction with cellular regulatory proteins and by its effect on cellular transcription. In this review, we discuss clinical aspects of PML, the ability of JCV to induce tumors in animal models, and the ability of JCV T-antigen to alter cellular function in vitro. PMID- 9852279 TI - Retinal pigment epithelial cell-based gene therapy against hemoglobin toxicity. AB - To determine if overexpression of the human heme oxygenase (HO-1) protects retinal pigment (RPE) cells from hemoglobin toxicity, a human RPE cell line was infected by an adenoviral vector containing the HO-1 (Ad-HO-1) gene or transfected with a plasmid containing the cytomegalovirus promoter and HO-1 cDNA (pRc/CMV-HO-1) complexed to cationic liposomes. Phase contrast microscopy and acid phosphatase activity were examined to insure homogeneity of the cell line. Mitochondrial cytochrome and microsomal heme content were measured in both transduced and control cells. RPE cells were then challenged with hemoglobin and their viability estimated. We determined that cells transfected with Ad HO-1 overexpressed HO-1 compared to control cells: HO-1 mRNA levels were increased 3 fold within 3 days, decreasing in 7 days. In addition, we permanently transfected RPE cells with HO-1 gene. Transfected cell clones selected for neomycin resistance had elevated levels of HO activity 3-fold higher than control. Transfected cells exposed to hemoglobin had a survival rate of 93%; non transfected cells had a 65-75% rate of survival. Transfected cells overexpressing HO-1 proved highly viable when challenged with hemoglobin. HO-1 appears to be an important component of the cellular anti-oxidant defense mechanisms against hemoglobin toxicity. However, the choice of transient or permanent expression of HO-1 against hemoglobin toxicity and hemorrhage needs to be further evaluated. PMID- 9852280 TI - Identification of a novel Rac3-interacting protein C1D. AB - Rac3 is a small GTPase of the Rho family, members of which have been implicated in tumorigenesis, cell growth/death and organization of the actin cytoskeleton. Many Rac-interacting proteins or effectors identified to date have a role in cytoskeletal organization. Using the yeast two-hybrid system, we have isolated a novel Rac3-interacting protein from a human placenta cDNA library which has no homology to other previously identified Rac-interacting proteins. Sequence analysis revealed that this protein is C1D, the human homolog of the murine SUN CoR protein which acts as a corepressor for the thyroid hormone receptor. In yeast cells, C1D binds to constitutively activated but not to GDP-bound Rac3. When coexpressed with Rac3 in COS cells, C1D complexed with constitutively active Rac3 but not with wild-type Rac3, demonstrating that C1D-Rac3 interactions take place in vivo in mammalian cells and that C1D appears to be an effector of Rac3. The C1D gene was mapped to human chromosome 2, which frequently shows deletions in human follicular thyroid carcinomas. PMID- 9852281 TI - Retrovirus-mediated gene therapy for human hepatocellular carcinoma transplanted in athymic mice. AB - Gene therapy using a retrovirus vector carrying herpes simplex virus thymidine kinase gene under the control of the 0.3-kb human alpha-fetoprotein (AFP) gene promoter (LNAF0.3TK virus) in combination with ganciclovir (GCV) treatment was performed in athymic mice harboring AFP-producing HuH-7 human hepatoma cells. GCV treatment resulted in pronounced growth inhibition of the virus-infected HuH-7 xenograft in mice, but did not affect growth of the parental xenograft. These results indicate that the AFP gene promoter sequence allows enough therapeutic gene expression to induce the GCV-mediated cytotoxicity in vivo in AFP-producing human hepatoma cells. PMID- 9852282 TI - Excitation-contraction-relaxation cycle: role of Ca2+-regulatory membrane proteins in normal, stimulated and pathological skeletal muscle (review). AB - Extremely large protein complexes involved in the Ca2+-regulatory system of the excitation-contraction-relaxation cycle have been identified in skeletal muscle, i.e. clusters of the Ca2+-binding protein calsequestrin, apparent tetramers of Ca2+-ATPase pump units and complexes between the transverse-tubular alpha1 dihydropyridine receptor and ryanodine receptor Ca2+-release channel tetramers of the sarcoplasmic reticulum. While receptor interactions appear to be crucial for signal transduction during excitation-contraction coupling, avoidance of passive disintegration of junctional complexes and stabilization of receptor interactions may be mediated by disulfide-bonded clusters of triadin. Oligomerization of Ca2+ release, Ca2+-sequestration and Ca2+-uptake complexes appear to be an intrinsic property of these muscle membrane proteins. During chronic low-frequency stimulation, the expression of triad receptors is decreased while conditioning has only a marginal effect on Ca2+-binding proteins. In contrast, muscle stimulation induces a switch from the fast-twitch Ca2+-ATPase to its slow twitch/cardiac isoform. These alterations in Ca2+-handling might reflect early functional adaptations to electrical stimulation. Studying Ca2+-homeostasis in transformed muscles is important regarding the evaluation of new clinical applications such as dynamic cardiomyoplasty. Studies of Ca2+-handling in skeletal muscle fibers have not only increased our understanding of muscle regulation, but have given important insights into the molecular pathogenesis of malignant hyperthermia, hypokalemic periodic paralysis and Brody disease. PMID- 9852283 TI - Cationic trypsinogen produced by human pancreatic ductal cancer has the characteristics of spontaneous activation and gelatinolytic activity in the presence of proton. AB - We examined whether human pancreatic ductal cancer cells express and secrete pancreatic cationic trypsinogen in vitro which can be spontaneously converted into active trypsin at acidic pH (pH 4.5-5. 5), in contrast to anionic trypsinogen. Cationic trypsinogen expression at the mRNA level was observed in differentiated Capan-1 and BxPC-3 cell lines. However, expression was not detected in either poorly-differentiated Panc-1 or undifferentiated MIAPaCa-2 cell line. The gelatinolytic activity of the activated form of trypsinogen in each conditioned medium in the presence of enterokinase (1.0 microg/ml) (a band with a molecular weight of approximately 23 kDa) corresponded well to the level of cationic trypsinogen mRNA. The spontaneous activation of trypsinogen also was observed by gelatin zymography of the acid-loaded conditioned medium (pH 5.5). These findings suggest that trypsinogen produced by human pancreatic ductal cancer has the characteristics of spontaneous activation and gelatinolytic activity in the presence of proton. PMID- 9852284 TI - Conjugate chemistry, iodination and cellular binding of mEGF-dextran-tyrosine: preclinical tests in preparation for clinical trials. AB - A conjugate with specific binding to the epidermal growth factor receptor, EGFR, and of interest for clinical tests was prepared using mouse epidermal growth factor, mEGF, and dextran. The mEGF was first coupled to dextran by reductive amination in which the free amino group on the N-terminal of mEGF was reacted with the aldehyde group on the reductive end of the dextran chain. The end-end coupled intermediate was further activated by the cyanopyridinium agent CDAP and tyrosines introduced to the dextran part of the conjugate. The mEGF-dextran tyrosine conjugate was, with high efficiency, iodinated with the chloramine-T method. Approximately 25-35% of the radioactivity could be removed from the conjugate after exposure to protease K while 65-75% of the radioactivity could be removed after exposure to dextranase. Thus, the largest amount of the iodine was on the dextran part of the conjugate. The iodinated mEGF-dextran-tyrosine had EGFR specific binding since the binding to an EGFR rich human glioma cell line could be displaced by an excess of non-radioactive mEGF. The conjugate was to a large extent internalized in these cells and the administrated radioactivity was thereby retained inside the cells for at least up to 50 h. PMID- 9852286 TI - ATP synthesis and export in heart left ventricle mitochondria from spontaneously hypertensive rat. AB - Use was made of mitochondria isolated from heart left ventricles of either spontaneously hypertensive or age-matched Wistar-Kyoto rats used as a control to find out whether hypertrophy (5-week-old rats) or hypertrophy/hypertension (24 week-old rats) can cause change in the mechanisms by which ATP is synthesised via ATP synthase and subsequently exported via the ADP/ATP translocator outside mitochondria. To do this, photometric measurements were made of the rate of ATP appearance in the extramitochondrial phase, which occurs as a result of ADP addition to mitochondria. In mitochondria from spontaneously hypertensive rats deficit of ATP production was found dependent on changes in the KmADP and Vmax values of both the ADP/ATP translocator and the ATP synthase. The ADP/ATP translocator was found to determine the rate of ATP production outside mitochondria in all the tested samples. In an initial investigation carried out to ascertain how cell ATP deficit can be counterbalanced, an increase in both adenylate kinase and creatine kinase activities was found in both hypertrophy and hypertrophy/hypertension. A possible increase in anaerobic glycolysis was also suggested by the increased lactate dehydrogenase activity. PMID- 9852285 TI - Determination of p53 protein and high-risk human papillomavirus DNA in carcinomas of the renal pelvis and ureter. AB - The presence and distribution of high-risk human papillomavirus (HPV) DNA type-16 and -18 or overexpression of the p53 protein were determined in 31 patients with renal pelvic and ureteral carcinoma in Saga prefecture of Japan, consisting of 28 transitional cell carcinomas (TCCs) and 3 squamous cell carcinomas (SCCs). In situ hybridization with DNA probes for HPV-16 and -18 was performed as well as immunohistochemical techniques using antibody to p53 protein. Fresh tumor tissues from 8 patients were also studied for p53 mutations in exons 5 through 8 by direct sequencing. Of 28 TCC patients at Saga prefecture in Japan, 10 tested were positive for HPV DNA, 5 for HPV type-16, 4 for HPV type-18 and one doubly positive case. Of the remaining 18 tumors, seven showed positive nuclear staining in cancer cells with p53 antibody. One of the 3 patients with SCCs also revealed the nuclear positivity of the cancer cells with p53 antibody but not with HPV infection. p53 point mutation was detected in 2 cases also showing p53 immunopositivity. Overexpressed p53 protein was statistically more common in invasive and non-papillary (p<0.05) and in high-grade TCCs (p<0.05). These findings suggest that HPV infection, and expression (mutation) of p53 protein may be intimately related to the pathogenesis of urothelial tumor cell growth and progression. PMID- 9852287 TI - A novel nuclear substructure, ND10: distribution in normal and neoplastic human tissues. AB - ND10 are recently characterized nuclear domains that are composed of 0.5 microm sized, precisely circumscribed dots in cultured human cell lines. To investigate the distribution and number of ND10 on various types of normal and neoplastic human tissues, we carried out immunostaining and immunoprecipitation analyses with monoclonal antibodies 138 and 1150. The number of ND10 varied from 1 to 10 or more in various tissues as did their size. ND10 were diffusely located in early embryonic and normal tissues, except for the exocrine and endocrine cells of the pancreas and for hepatocytes. In normal squamous mucosa, basal cells had more ND10 than did differentiated superficial squamous cells. The number and size of ND10 were markedly increased in malignant neoplasms but were similar in benign tumors and corresponding normal tissues. Sex hormone-related normal tissues, such as the endometrium or myometrium, and neoplasms strongly stained for ND10. The distribution pattern of ND10 in human tissues indicates that they are conserved nuclear substructures that are closely associated with cellular differentiation, hormonal stimulation, and oncogenesis. PMID- 9852288 TI - Black tea theaflavins induce programmed cell death in cultured human stomach cancer cells. AB - The exposure of human stomach cancer KATO III cells to black tea theaflavin extract, free theaflavin, and theaflavin digallate that are main components of the extract, led to both growth inhibition and the induction of programmed cell death (apoptosis). Morphological changes showing apoptotic bodies were observed in the cells treated with black tea theaflavin extract, theaflavin and theaflavin digallate. The fragmentations by these theaflavin compounds of DNA to oligonucleosomal-sized fragments that are characteristics of apoptosis were observed to be concentration- and time-dependent. These data suggest that drinking of black tea in large amounts is recommended to protect humans from stomach cancer. PMID- 9852289 TI - Lymph nodes and human tumors (review). AB - This review examines the crucial role of regional lymph nodes (RLN) in defense against tumor progression. RLN are one of the first major components of the immune system to come into contact with tumor cells or tumor-cell products and are important in the generation of tumor-directed immune responses. Involvement of RLN by tumor cells is a prognostic index of survival and a biological indicator of a more distant metastatic disease. Enlargement of lymph nodes as a consequence of the increase in the number of lymphoid cells, is a common finding in humans. These changes of cellular organization display the most decisive evidence of the existence of an immune response within a draining lymph node. The variety of cells mediating immune response to tumors are summarized briefly. The lymphocyte subpopulations involved reflect the nature of the response and may determine the outcome of the tumor-host interaction. The composition of the lymphocyte subpopulations can be recognized in tumor-draining lymph nodes by distinctive surface-membrane markers assessable by flow cytometry. In human patients with solid tumors limited quantification of the lymphocyte subpopulations within RLN has been carried out using this technique and the results indicated that an increase in B lymphocytes in tumor-reactive lymph nodes is marked in the adenocarcinomas (colon and breast) while in other tumors, such as melanoma and squamous cell carcinoma, this increase in B lymphocytes is less pronounced. The increased number of B lymphocytes in the reactive lymph nodes suggests the existence of an immune response involving interactions between T and B cells. B lymphocytes expression of CD80 appears to increase in some reactive lymph nodes to adenocarcinomas, possibly indicating the state of activation of CD80+ B cells, and their role as antigen-presenting cells. Any improvement in the antitumor activity of RLN would be important in the immunotherapy of cancer patients. The ability to generate a large number of tumor-reactive T lymphocytes is a critical requirement for adoptive immunotherapy. Tumor-draining lymph nodes (TDLN) are an excellent source of tumor-reactive T lymphocytes and the adoptive transfer of these cells is capable of mediating the regression of tumors established both in the lung and in the brain. Although cancers elicit a vigorous immune response during the early part of their growth, the immune response is soon downregulated, permitting progressive cancer growth. Furthermore, there are date suggesting the existence of immunosuppressive mechanisms within RLN in the antitumor response. However, there are no yet conclusive data concerning the characteristics of the response or its effectiveness. PMID- 9852290 TI - Epidermal growth factor downregulates the expression of SH-PTP2. AB - Protein phosphorylation/dephosphorylation on tyrosine residues are regulated by tyrosine kinases/phosphatases. The tyrosine phosphatase SH-PTP2 (PTP1D, PTP2C) interacts physically through its SH2 domain with phosphorylated epidermal growth factor receptor (EGFR). In KB cells, an oral epidermoid carcinoma, high epidermal growth factor (EGF) concentrations (10-9, 10-8 and 10-7 M) downregulate its receptor for the duration of the incubation with EGF. Thus, it was hypothesized that in KB cells, SH-PTP2 expression would also be downregulated by high EGF concentrations. This hypothesis was investigated by incubating the KB cells with increasing concentrations of EGF (0, 10-11, 10-10, 10-9, 10-8, 10-7 M) and by evaluating the expression of SH-PTP2 protein under these conditions. This study showed that EGF 10-7 and 10-8 M significantly decreased SH-PTP2 protein expression compared to controls. EGF 10-10 and 10-11 M did not change the expression of SH-PTP2 protein. We conclude that high EGF concentrations downregulate the expression of SH-PTP2 protein. PMID- 9852291 TI - Prothymosin alpha1 antagonizes the inhibitory effects of transforming growth factor-beta1 on the adhesion of peripheral blood lymphocytes to human umbilical vein endothelial cells. AB - We investigated the in vitro effects of both prothymosin alpha1 (Pro alpha1) and transforming growth factor-beta1 (TGF-beta1) on the adhesion of peripheral blood lymphocytes (PBL) and on the expression of the adhesion molecules, endothelial leukocyte adhesion molecule-1 (ELAM-1), intracellular adhesion molecule-1 (ICAM 1) and vascular cell adhesion molecule-1 (VCAM-1) on human umbilical vein endothelial cells (HUVECs). TGF-beta1 moderately but significantly decreased PBL binding to interleukin-1 (IL-1)- stimulated HUVECs. However, Pro alpha1 in combination with TGF-beta1 completely restored the TGF-beta1 mediated effects. Other thymic peptides tested were ineffective. On HUVECs, TGF-beta1 diminished ICAM-1 and VCAM-1 expression, leaving ELAM-1 unchanged. Pro alpha1 in combination with TGF-beta1 showed no significant effects on ELAM-1 expression, but antagonized the TGF-beta1-induced decrease of ICAM-1 and VCAM-1 expression on activated HUVECs. PMID- 9852292 TI - Effect of dietary phytochemicals on cancer development (review) AB - Vegetables, fruits, and whole grains contain a wide variety of phytochemicals that have the potential to modulate cancer development. There are many biologically plausible reasons why consumption of plant foods might slow or prevent the appearance of cancer. These include the presence in plant foods of such potentially anticarcinogenic substances as carotenoids, chlorophyll, flavonoids, indole, isothiocyanate, polyphenolic compounds, protease inhibitors, sulfides, and terpens. The specific mechanisms of action of most phytochemicals in cancer prevention are not yet clear but appear to be varied. Considering the large number and variety of dietary phytochemicals, their interactive effects on cancer risk may be extremely difficult to assess. Phytochemicals can inhibit carcinogenesis by inhibiting phase I enzymes, and induction of phase II enzymes, scavenge DNA reactive agents, suppress the abnormal proliferation of early, preneoplastic lesions, and inhibit certain properties of the cancer cell. PMID- 9852293 TI - Bcl-2 protein and apoptosis in pediatric acute lymphoblastic leukemia. AB - Many chemotherapeutic agents are thought to exert their genotoxic effects through induction of programmed cell death (PCD) (apoptosis) in tumor cells. The bcl-2 is an anti-apoptotic oncoprotein and can confer a survival advantage to tumor cells by preventing apoptosis. Overexpression of bcl-2 may therefore be implicated in resistance to chemotherapy. We studied the significance of bcl-2 expression and the PCD index in pediatric acute lymphoblastic leukemia. Evaluation of bcl-2 by immunocytochemistry and PCD by an enzymatic end labelling technique using biotin dUTP was carried out in a total of 55 cases and 40 controls. Bcl-2 was found to be expressed in 47% (26/55) of the acute lymphoblastic leukemia cases. The positive cells varied from 0-49% among individual samples. Pre-treatment (spontaneous) apoptosis was observed in 62% (34/55) cases. The mean pre-treatment PCD index was 8.27 1.3%, while the median PCD index was 5. The PCD value for the leukemic samples analyzed were then classified as either high apoptosis values ( 5) and low apoptosis values (<5). PCD index was high in 53% (29/55) and low in 47% (26/55). However, 23% (13/55) of cases did not show presence of either apoptosis or bcl-2. There was no association between clinical and laboratory parameters with the apoptotic index or bcl-2 protein expression. However, evaluation of apoptotic index and bcl-2 expression on day 7 of induction chemotherapy showed a borderline correlation between these markers and initial WBC count, presence of mediastinal mass and hepatosplenomegaly. Follow-up of these patients is being done to look for any association between treatment response and apoptosis. PMID- 9852294 TI - Keratinocyte growth inhibition by streptococcal proteins. AB - M proteins are receptor proteins and one of the virulence factors of streptococci. M proteins seem to play a role in inflammatory skin disorders such as psoriasis. It is however unknown whether M proteins have a direct influence on proliferative activity of human keratinocytes. In the present study human HaCaT keratinocytes were exposed to M proteins (M1, M3, M5, M12) and the proliferative and proinflammatory response was analyzed. We found a dose-dependent inhibition of keratinocyte proliferation with crude extract of strain M3 4/55. Following affinity chromatography we found inhibitory activity for keratinocyte proliferation with a maximum of 80% at 10-8 M in the M protein. Additionally tested M1 protein preparation showed an inhibitory activity of 55% whereas other M preparations (5 and 12) did not show any effect. In supernatants from HaCaT cultures IL-1alpha, IL-1beta, IL-6, IL-8, TNFalpha and ICAM-1 were measured by ELISA. The levels of IL-8 were high and TNFalpha was upregulated, whereas ICAM-1 was decreased from around 20 ng/ml to almost zero. In contrast to the streptococcal-derived M3 protein preparation the recombinant M3 did not interfere with the proliferation of HaCaT cells. Because neither recombinant M3 protein nor M3 protein purified by ion exchange chromatography on a Q-resource column had any antiproliferative activity on keratinocytes we suggest, that a component different from M3 protein was responsible. PMID- 9852295 TI - Specificity of polyclonal anti-p53 IgG for isolation of the soluble p53 antigen from human serum. AB - The possibility to use anti-p53 IgG for isolation of the soluble p53 antigen as a serological tumor marker has been shown in our previous studies. In order to prove the specificity of such IgG, we compared the effectiveness of columns with anti-p53 IgG and IgG isolated from non-treated rabbits (regular IgG). The gel fiberglass (GFG) columns for affinity chromatography were prepared separately with both types of IgG. The same serum was percolated simultaneously through both columns and the results of elution were compared. The total concentration of tumor-associated antigens (TAA) eluted from the serum of cancer patients was similar in both TAA mixtures isolated either with anti-p53 IgG or with the regular IgG. Differences are manifest in the content of total proteins and amount of each eluted protein: a mixture eluted with the regular IgG contains several proteins whereas the anti-p53 IgG isolated only two proteins, p64 and p53. The amount of the soluble p53 antigen isolated from the cancer serum was significantly higher when it was isolated with anti-p53 IgG. The method developed in our laboratory was shown to be highly specific both to isolate proteins related to cancer (p53) and non-cancer disorders (p64). Data presented in this report show that such specificity can be achieved only if the anti-p53 IgG is used: the regular IgG obtained from non-treated animals isolate many proteins among which the concentration of specific p53 protein is lost. PMID- 9852296 TI - Association of N-acetyllactosamine with tumor progression in human breast cancer: a study using a 16 kDa chick embryo lectin. AB - This study documents modifications in the expression and the cellular distribution of binding sites for a 16 kDa chick embryo lectin (CL16-BS) in breast cancer. This lectin binds preferentially to terminal and penultimate N acetyllactosamine residues (Galbeta1-4GlcNAc). BS density and distribution, studied by lectin binding followed by indirect immunofluorescence, were compared in normal breast tissues and 45 invasive carcinomas (lobular and ductal). Increased number of fluorescent epithelial cells (ETC+) were observed in normal ducts adjacent to lobular carcinomas and in tumors from both types when compared to normal glands. In ductal carcinomas, a significant diminution of ETC+ percentage was observed in the highest anatomopathological SBR grades: 32.7% in grade III, 80.8% in grade II and 66.5% in grade I (p<0.001). For both lobular and ductal carcinomas, ETC+ percentages were also positively correlated with low versus high MSBR grades (p<0.002). The subcellular distribution of CL16-BS varied according to the tumor type and/or the histological grades. It was mostly membrane-associated in low SBR and MSBR grades (p<0.001 and p<0.01, respectively) and cytoplasm-associated in high grades (p<0.02 and p<0.05). Some of these parameters were also correlated with certain other clinicopathological factors, such as tumor size (p<0.02), high S-phase cell fraction (p<0.04 and p<0.03) and low density estrogen receptors (p<0.05). Diminution in CL16-BS density and cytoplasmic versus membrane localization may be considered as indicators of tumor progression but not of metastasis. PMID- 9852297 TI - Oxidation, tyrosine nitration and cytostasis induction in the absence of inducible nitric oxide synthase. AB - In the present study, we evaluated the impact of the lack of the gene for inducible nitric oxide synthase (iNOS) on oxidation, tyrosine nitration and cytotoxicity reactions triggered by immunostimulation. In mice injected with E. coli endotoxin (bacterial lipopolysaccharide, LPS, 50 mg/kg i.p.), there was a significant increase in the degree of oxidation of dihydrorhodamine 123 to rhodamine 123. This response was attenuated by inhibition of NO biosynthesis with NG-methyl-L-arginine (L-NMA, 30 mg/kg i.p.). In mice lacking functional iNOS gene (iNOS knock-out mice), the degree of the LPS-induced, L-NMA inhibitable increase in dihydrorhodamine oxidation was decreased, but not completely abolished. LPS stimulation induced a marked increase in the immunoreactivity for nitrotyrosine (an indicator of peroxynitrite formation), as measured in the aorta and lung. An L-NMA inhibitable increase in nitrotyrosine staining induced by LPS was also observed in the tissues of the iNOS knockout animals. LPS treatment induced the appearance of DNA single strand breakage and a suppression of mitochondrial respiration in peritoneal macrophages ex vivo. A significant degree of LPS induced DNA single strand breakage and suppression of mitochondrial respiration was still observed in the peritoneal macrophages obtained from the iNOS knockout animals. Macrophages from wild-type mice stimulated with LPS and interferon-gamma suppressed the proliferation of various target cells (P815 mastocytoma, L929 fibrosarcoma and embryonic lung fibroblast cell line): this effect was abolished by in vitro treatment with L-NMA (1 mM). Macrophages from the iNOS knockout animals exhibited a reduced degree of target cell cytostatic activity. The remainder of the cytostasis in iNOS knockout macrophages was abolished by preventing cell contact and neutralizing tumor necrosis factor a. The present results demonstrate that the lack of iNOS gene does not fully abolish oxidation, tyrosine nitration and cytostatic activity in response to immunostimulation. The current findings may have implications for the development of NO-based approaches for the experimental therapy of inflammation. PMID- 9852298 TI - The presenilin 2 loop domain interacts with the mu-calpain C-terminal region. AB - Presenilin 2 (PS2) is a gene responsible for the early-onset familial Alzheimer's disease (AD). PS2 mutations are considered to be closely related to the pathogenesis of AD. We screened for proteins that interact with PS2 to understand its pathological and physiological functions. Using the PS2 loop domain as the bait, the yeast two-hybrid system was used for screening, and mu-calpain was identified as a PS2 binding protein. In COS-1 cells, the interaction of PS2 with mu-calpain was confirmed by immunoprecipitation. These results suggested that PS2 and mu-calpain interact with each other, and might regulate each other's functions. PMID- 9852299 TI - Characterization of the breakpoints in unbalanced t(5;11)(p15;p15) constitutional chromosome translocations in two patients with beckwith-wiedemann syndrome using fluorescence in situ hybridisation. AB - Although the majority of patients with Beckwith-Wiedemann syndrome (BWS) have a normal karyotype, the study of those rare patients with a cytogenetic abnormality has given considerable insight into the genetics of this condition. The karyotypic abnormalities found include partial chromosome duplications of paternal origin and maternally derived translocations which usually involve the 11p15 region and provide one of the lines of evidence for the location of the BWS gene(s). Because the extent of the duplicated region in these patients is variable, the phenotypic expression of BWS is presumably due to the presence of a common duplicated region. Two unrelated patients with BWS were both noted to have a similar unbalanced t(5;11)(p15;p14) translocation. The parents in both families were unaffected but both fathers carried a balanced translocation involving the same chromosomes. Since the extent and nature of the duplication apparently determines the complex phenotypes seen in these patients, we undertook a detailed analysis of the extent of the triplicated region using fluorescent in situ hybrisation (FISH). Despite having markedly different phenotypes and presenting in disimilar ways the two patients had apparently identical duplication breakpoints. PMID- 9852300 TI - CD46 (membrane cofactor protein of complement, measles virus receptor): structural and functional divergence among species (review). AB - Human CD46 was identified as a complement regulator and was later shown to be a measles virus receptor. The ubiquitous distribution profile of CD46 accounted for systemic measles infection and general protection of host tissue/organs from autologous complement. A similar ubiquitous distribution was observed for swine and simian CD46 homologues based upon subsequent cDNA cloning and Northern analysis, reinforcing the roles of CD46. In contrast, recent cDNA cloning and distribution analyses of murine and guinea-pig CD46 revealed the predominant expression of these rodent CD46 homologues in the testis, especially in mature testicular germ cells. These results do not support the established functions of human CD46 but support the hypothesis that CD46 on sperm serves as a fertilization-related adhesion molecule toward eggs. Here, we review the structure, function and distribution of human CD46 and discuss the possible differences between human CD46 and its homologues recently cloned from a variety of non-human primates and other animals. PMID- 9852301 TI - Inhibition in a microgravity environment of the recovery of Escherichia coli cells damaged by heavy ion beams during the NASDA ISS phase I program of NASA Shuttle/Mir mission no. 6. AB - We participated in a space experiment, part of the National Space Development Agency of Japan (NASDA) Phase I Space Radiation Environment Measurement Program, conducted during the National Aeronautics and Space Administration (NASA) Shuttle/Mir Mission No. 6 (S/MM-6) project. The aim of our study was to investigate the effects of microgravity on the DNA repair processes of living organisms in the in orbit. Heavy ion beam radiation- or c-irradiation-damaged biological samples of Escherichia coli and the radioresistant bacterium Deinococcus radiodurans were prepared and placed in a biospecimen box, which was loaded into the RRMD III sensor unit of the Space Shuttle. Two identical sets of samples were left in the Spacehab's Payload Processing Facility (SPPF) in Florida, USA, as a control. (flight No. STS-84) was launched from NASA John F. Kennedy Space Center (KSC) in Florida, USA, on May 15, 1997. The mission duration was 9.22 days. An astronaut activated the biological samples in the biospecimen box in the Spacehab during orbit in order to start repair of the DNA damaged by heavy ion beams or c irradiation and the samples were incubated for 19 h 35 min at about 22uC, the cabin temperature. The control specimens in the SPPF were subjected to the same treatment under terrestrial gravity. After returned to earth, we investigated cell recovery by comparing the repair of the radiation-damaged DNA of E. coli and D. radiodurans in the microgravity environment in space with that on Earth. The results indicated that the DNA repair process of E. coli, but not of D. radiodurans, cells was inhibited in a microgravity environment. PMID- 9852302 TI - p53 alteration, proliferating cell nuclear antigen, and nucleolar organizer regions in thymic epithelial tumors. AB - We examined p53 protein expression, p53 gene mutation, proliferating cell nuclear antigen (PCNA), and argyrophilic nuclear organizer regions (AgNOR), in 30 patients with surgically-treated thymic tumors (26 thymoma and 4 thymic carcinoma cases). p53 expression ratio with DO-1 was divided as p53 negative (0% positivity), low expressor (<10% positivity), high expressor (>10% positivity). The incidence of p53 low and high expressor in thymoma were 19% (5/26) and 8% (2/26), respectively. p53 immunopositivity in thymoma was significantly correlated with PCNA labeling index (LI). p53 expression ratio in invasive thymoma (33%) tended to be higher than that in non-invasive thymoma (18%). p53 expression was detected in one of the thymic carcinoma. There were no p53 gene mutations in 15 invasive thymoma, although one of four (25%) thymic carcinomas showed two point mutations. p53 gene alterations seem to be associated with malignant activity of tumor cells, and therefore detection of p53 gene mutations is useful as a diagnostic factor. PMID- 9852303 TI - Signal transduction in arteriosclerosis: mechanical stress-activated MAP kinases in vascular smooth muscle cells (review). AB - Vascular smooth muscle cell (SMC) proliferation is a key event in the development of (spontaneous) atherosclerosis, hypertension-related arteriosclerosis, angioplasty-induced restenosis and venous bypass graft arteriosclerosis. Many factors or environmental stimuli are believed to be responsible for SMC growth or hypertrophy in the vessel wall. How these environmental stimuli or signals applied onto the surface of SMCs are transduced into the cell nucleus resulting in quantitative and qualitative changes in gene expression in SMCs of arterial walls is largely unknown. Mitogen-activated protein (MAP) kinases are rapidly activated in cells stimulated with various extracellular signals by dual phosphorylation of tyrosine and threonine residues. They are thought to play a pivotal role in transmitting transmembrane signals required for cell growth and differentiation. Recent studies have focused on the signalling events in vascular tissues in vivo and in cultured SMCs in vitro. It has been demonstrated that acute hypertension and angioplasty rapidly induced MAP kinase activation in the arterial wall. Kinase activation is followed by an increase in c-fos and c-jun gene expression and enhanced transcription factor AP-1 DNA-binding activity. A similar MAP kinase activation can be mimicked in in vitro cultured SMCs stimulated by either shear stress or cyclic strain stretch, suggesting direct effects of mechanical force. Interestingly, physical forces rapidly resulted in phosphorylation of platelet-derived growth factor (PDGF) receptor, an activated state, in cultured SMCs. Thus, mechanical stresses may directly perturb the cell surface or alter receptor conformation, thereby initiating signalling pathways usually used by growth factors. These findings have significantly enhanced our knowledge concerning the pathogenesis of arteriosclerosis and provide a basis for therapeutic intervention on vascular diseases. PMID- 9852304 TI - Telomerase activity in malignant and benign bladder conditions. AB - This study investigated whether the enzyme telomerase is active in bladder tumours, whether there is any correlation between activity and grade, and whether the enzyme is expressed in non-malignant conditions. Fifty-two patients undergoing cystoscopy or TURBT at a district general hospital were included, 25 with current bladder tumours, 13 with previous but no current tumours, and 14 with non-malignant pathology. Specimens were analysed by the telomerase repeat amplification protocol (TRAP assay), a highly-sensitive polymerase chain reaction (PCR)-based assay, and a commercially-available ELISA kit. Telomerase activity was detected in 80% of bladder tumours, more frequently in moderate- or poorly differentiated (93%) than well-differentiated (56%) tumours. Activity was not uniform across individual tumours. Telomerase was also frequently (61%) detected in inflammatory lesions found in patients being followed up for previous bladder tumours, and in two (14%) patients with benign pathologies. In conclusion, telomerase was frequently but not uniformly detected in bladder tumours; its presence was not specific to malignancy. There is a possible correlation between tumour grade and telomerase activity. PMID- 9852305 TI - Expression of somatostatin receptor genes and acetylcholine receptor development in rat skeletal muscle during postnatal development. AB - Our laboratory reported previously that somatostatin (SST) is transiently expressed in rat motoneurons during the first 14 days after birth. We investigated the possibility that the SST receptor (SSTR) is expressed in skeletal muscle. We found that two of the five subtypes of SSTR (SSTR3 and SSTR4) are expressed in skeletal muscle with a time course that correlates with the transient expression of SST in motoneurons. In addition, SSTR2A is expressed from birth to adulthood in skeletal muscle. Both SSTR2A and SSTR4 are also expressed in L6 cells, a skeletal muscle cell line. Somatostatin acting through its receptors has been shown to stimulate tyrosine phosphatase activity in a number of different tissues. We found that several proteins (50, 65, 90, 140, 180 and 200 kDa) exhibited a reduced degree of tyrosine phosphorylation following SST treatment. Inhibition of tyrosine phosphatase activity with sodium orthovanadate increased expression of the nicotinic acetyl-choline receptor (nAChR) epsilon subunit mRNA by three fold. Somatostatin reversed the elevated epsilon mRNA following orthovanadate treatment. These findings show that SSTR is expressed in skeletal muscle and that SST acting via the SSTR regulates tyrosine phosphorylation and expression of the epsilon subunit of the AChR in the rat skeletal muscle. PMID- 9852307 TI - p185 overexpression in 220 samples of breast cancer undergoing primary surgery: comparison with c-erbB-2 gene amplification. AB - In breast cancer, DNA amplification of the oncogene c-erbB-2, encoding for the p185 protein, is associated with a poor prognosis. A retrospective study on a population of 220 cases of primary breast cancer permitted a quantitative measure of p185 oncoprotein overexpression by an immunoenzymetric assay and the determination of c-erbB-2 amplification by the Southern blot method. A correlation existed between the two measurements (r=0.85) using the double cut off: DNA 2 copies and p185 400 U/mg protein, and only 2.7% of the cases were discordant. 13.2% of the tumors showed p185 overexpression. The percentage of tumors overexpressing p185 was significantly different between the groups with amplified and non-amplified c-erbB-2. We observed a significant correlation between p185 levels and tumor grade (p=0.03), and an inverse correlation with hormonal receptors (p=0.0001). The p185 assay could be an additional prognostic factor to better define patient subgroups with node negative, grade II, and positive or negative hormonal receptors. PMID- 9852306 TI - Abnormal cortical development; towards elucidation of the LIS1 gene product function (review). AB - Lissencephaly is a relatively common brain malformation. Lissencephaly type 1 is characterized by the smooth appearance of the cortex and the presence of four abnormally positioned layers instead of the normal six. Lissencephaly is considered to be an abnormality in neuronal migration. The gene mutated in type 1 lissencephaly was cloned by us and designated LIS1. Recently, several genes involved in cortical development have been cloned in the mouse. In human an additional X-linked lissencephaly gene has been identified. We summarize here our current knowledge on the LIS1 gene and its function. It has been identified as a non-catalytic subunit of PAF-acetylhydrolase, a heterotrimeric enzyme which inactivates the platelet-activating factor (PAF). In addition, we have demonstrated that LIS1 interacts with tubulin, and affects the dynamics properties of microtubles. LIS1 contains seven WD repeats and may structurally resemble the beta-subunit of heterotrimeric G proteins. Interestingly, the catalytic subunit of PAF-acetylhydrolase was found to resemble the alpha subunit of heterotrimeric G proteins. We raise the possibility that LIS1 is part of an intracellular signaling pathway involved in neuronal migration. PMID- 9852308 TI - Analysis of kappa light chain contribution to anti-DNA antibody activity of a human VH4-21-encoded monoclonal antibody (NE-1) by antibody-phage display technique. AB - Although contribution of light chain to DNA reactivity of some murine anti-DNA antibodies (Abs) has been demonstrated, similar studies on human anti-DNA Abs are limited. To investigate this contribution, we reproduced Fab molecules on the surface of phages from a human B cell line producing IgM anti-DNA monoclonal Ab (NE-1) by an Ab-phage display technique. Expressed Fab molecules (p4-1 clone) were similar to the parental mAb in their binding activities and idiotypic expression. We constructed a light chain shuffled library containing Vkappa genes derived from peripheral blood lymphocytes of a patient with systemic lupus erythematosus (SLE) in combination with the NE-1 heavy chain gene. After panning to ss- or dsDNA, 7 Fab-phage clones which showed significant bindings to ss- or dsDNA were isolated. Many other Fab-phage clones from the library did not bind to ss- nor dsDNA. Sequence analysis revealed that light chains of the 7 clones are derived from diverse Vkappa germline genes including rarely used ones such as the L5 and A30. Most of the Vk germline genes have been used for previously reported anti-DNA antibodies. These findings suggest that diverse Vkappa genes can pair with the NE-1 heavy chain for anti-DNA Ab activity. In addition, kappa light chains seem to modulate DNA binding activities in various ways. PMID- 9852309 TI - ATP in iron overload-induced intracellular calcium changes. AB - The cellular iron uptake from low molecular weight iron complexes (ferric citrate, ferric lactate and ferric ATP complex) is concentration-dependent, and only a small part of the iron penetrates the cell as shown by deferoxamine treatment. A threshold of iron concentration in the cell must be reached for the iron complex-induced increase of cellular Ca2+-uptake. ATP seems to play a key role in an iron translocation that enhances the effects of the iron complexes. A non-specific and competitive iron-binding by proteins seems to act as a buffer system that reduces the iron overload effects. Calcium channel blockers have no effects on the iron complex-cell interaction or iron-induced Ca2+-uptake modification. An iron complex concentration-dependent inhibition of the CaATPase activity, and its consequent Ca2+-extrusion impairment appear as the likely cause of calcium overload. The relevance of these findings in iron overload-induced pathologies is discussed. PMID- 9852310 TI - Human immunodeficiency virus type-1 transcription: role of the 5'-untranslated leader region (review). AB - Human immunodeficiency virus type-1 (HIV-1) transcription is dependent on the interaction of host-cell transcription factors with cis-regulatory DNA elements within the viral long terminal repeat (LTR). Much attention has focused on the series of sequence elements upstream of the transcriptional initiation site in the U3 region of the LTR including the Sp1 and NF-kappaB binding sites. Recent studies, however, demonstrate that the transcribed 5'-untranslated leader region (5'-UTR) also contains important transcriptional elements. These regulatory elements situated downstream of transcription interact with constitutive and inducible transcription factors, mediate transmission of cellular activation signals, and are important for efficient HIV-1 transcription and replication. The 5'-UTR contains binding sites for the transcription factors AP-1, NF-kappaB, NF AT, IRF, and Sp1. Mutations in these binding sites can interfere with the viral response to cell activation signals, decrease LTR transcription, and inhibit viral replication. The 5'-UTR also interacts with a specific nucleosome that is rapidly displaced during transcriptional activation of the latent provirus. We propose that the inducible transcription factor binding sites in the 5'-UTR comprise a downstream enhancer domain that can function independent of, or in concert with, the LTR promoter to rapidly increase latent proviral transcription in response to cell activation signals. In this review, we describe the host-cell transcription factors that interact with the 5'-UTR and discuss their role in the transcriptional regulation of HIV-1 gene expression. PMID- 9852311 TI - Antitumor effects elicited by antisense-mediated downregulation of the insulin like growth factor I receptor (review). AB - Antisense-mediated downregulation of the insulin-like growth factor I receptor results in massive apoptosis of tumor cells in vivo, leading to abrogation of tumorigenesis. In addition to the apoptotic effect, antitumor responses are elicited in syngeneic immunocompetent animals, protecting them from subsequent tumor challenge and causing regression of established tumors with no further recurrence. The mechanisms involved in the antitumor responses elicited in the animals following exposure to antisense cells are discussed, focusing on the immunogenicity of the antisense cells as well as the effectors that participate in these responses. PMID- 9852312 TI - Allelic status of 1p, 14q, and 22q and NF2 gene mutations in sporadic schwannomas. AB - Schwannomas are common benign tumours of schwann cell origin, frequently found in patients with neurofibromatosis type 2 (NF2). Inactivation of the NF2 tumour suppressor gene appears to be a molecular event responsible for the development of up to 60% of cases, but no data are available on other superimposed secondary or alternative molecular abnormalities in those schwannomas lacking NF2 gene inactivation. We analysed 23 sporadic schwannomas for mutations in the NF2 gene and for the allelic status at 1p, 14q and 22q, as alterations of these genomic regions appear to be related to tumour progression in meningiomas, another NF2 associated neoplasm. Nine samples displayed allelic losses for markers on chromosome 22, and deletions at 1p were detected in two. No case showed losses for 14q. Three tumours displayed NF2 gene mutations, at exons 2, 7 and 12. Our results confirm that inactivation of the NF2 gene is a primary event in schwannoma development, and provide data suggesting that allelic loss at 1p may contribute to the pathogenesis of a small subgroup of this histological tumour type. PMID- 9852313 TI - Cell biology and possible therapeutic applications of anti-CD3-activated killer-T cells (review). AB - Polyclonal T lymphocyte populations can be stimulated with anti-CD3 antibody to proliferate, secrete cytokines, and mediate MHC-unrestricted cytotoxic activity against a wide range of tumor target cells. Because anti-CD3-activated killer-T (AK-T) cells may be useful in the immunotherapy of human cancers, it is important to understand the signaling pathways and cell-surface structures involved in the induction and tumoricidal effector function of AK-T cells. Studies in the mouse model system have characterized the cytokines, signal transduction pathways, and costimulatory molecules involved in AK-T cell development. The recognition/adhesion and subsequent signaling events which lead to tumoricidal activity by mouse AK-T cells have also been defined. These findings, providing they translate accurately to the human system, may allow for the design of effective strategies to use AK-T cells for the treatment of human cancers. However, to date, the encouraging results obtained with anti-CD3 antibody/AK-T cell-based immunotherapies in mouse models of cancer have not been duplicated in clinical trials. The most likely explanation for this dis-appointing result is that tumor-reactive T lymphocytes in long term tumor-bearers fail to function correctly in the tumor microenvironment due to tumor-induced immune suppression and defects in key signal transduction molecules. It is clear that a detailed understanding of the inhibitory effect of established tumors on host T cells and the means to overcome tumor-induced immunosuppression are needed before anti-CD3 antibody/AK-T cell-based immunotherapies can be expected to succeed in the clinical setting. PMID- 9852314 TI - Microdialysis update: optimizing the advantages. AB - Microdialysis was introduced in the early 1970s as a method to measure dynamic release of substances in the brain (see Tossman & Ungerstedt, 1986). The technique has been refined over the past three decades due to the development of new materials for dialysis membranes and commercial availability of smaller, more consistently fabricated probes. A typical microdialysis probe consists of rigid metal concentric tubing with a semipermeable region at the tip (Fig. 1). Molecules of restricted size passively diffuse from the brain through the dialysis membrane into an infusion solution which is then directed out of the brain and collected in tubes for serial analysis of substance content. Probes are inserted into the brain region of interest, typically making lesions during their travel through the brain and at the sampling site. Once the trauma of insertion subsides, usually after an hour or so, probes collect substances released from axons projecting to dendrites and cell bodies of the targeted area. Substances surrounding the semipermeable region of the probe passively diffuse down a concentration gradient into the solution infused through the probe. Substance recovery from the brain decreases exponentially with faster infusion rates. A high precision infusion pump is critical for maintaining constant flow through the probes to ensure that altered substance content in the dialysates reflects changes in release by the brain and not variable diffusion gradients resulting from sporadic changes in flow rates through the probes. High performance liquid chromatography (HPLC) is commonly used to measure target substances in the dialysates, but other methods such as radioimmunoassay may be employed. The development of microbore columns for HPLC (Durkin et al. 1985) and their commercial availability by the mid 1990s has made it possible to accurately measure smaller amounts of substances in the dialysates. PMID- 9852315 TI - Changes in conformation of myosin heads during the development of isometric contraction and rapid shortening in single frog muscle fibres. AB - 1. Two-dimensional X-ray diffraction patterns were recorded at the European Synchrotron Radiation Facility from central segments of intact single muscle fibres of Rana temporaria with 5 ms time resolution during the development of isometric contraction. Shortening at ca 0.8 times the maximum velocity was also imposed at the isometric tetanus plateau. 2. The first myosin-based layer line (ML1) and the second myosin-based meridional reflection (M2), which are both strong in resting muscle, were completely abolished at the plateau of the isometric tetanus. The third myosin-based meridional reflection (M3), arising from the axial repeat of the myosin heads along the filaments, remained intense but its spacing changed from 14.34 to 14.56 nm. The intensity change of the M3 reflection, IM3, could be explained as the sum of two components, I14.34 and I14.56, arising from myosin head conformations characteristic of rest and isometric contraction, respectively. 3. The amplitudes (A) of the X-ray reflections, which are proportional to the fraction of myosin heads in each conformation, changed with half-times that were similar to that of isometric force development, which was 33.5 +/- 2. 0 ms (mean +/- s.d., 224 tetani from three fibres, 4 C), measured from the end of the latent period. We conclude that the myosin head conformation changes synchronously with force development, at least within the 5 ms time resolution of these measurements. 4. The changes in the X-ray reflections during rapid shortening have two temporal components. The rapid decrease in intensity of the 14.56 nm reflection at the start of shortening is likely to be due to tilting of myosin heads attached to actin. The slower changes in the other reflections were consistent with a return to the resting conformation of the myosin heads that was about 60 % complete after shortening of 70 nm per half-sarcomere. PMID- 9852316 TI - Effects of ivermectin and midecamycin on ryanodine receptors and the Ca2+-ATPase in sarcoplasmic reticulum of rabbit and rat skeletal muscle. AB - 1. Ryanodine receptor (RyR) Ca2+ channels in the sarcoplasmic reticulum (SR) of skeletal muscle are regulated by the 12 kDa FK506- (or rapamycin-) binding protein (FKBP12). Rapamycin can also activate RyR channels with FKBP12 removed, suggesting that compounds with macrocyclic lactone ring structures can directly activate RyRs. Here we tested this hypothesis using two other macrocyclic lactone compounds, ivermectin and midecamycin. 2. Rabbit skeletal RyRs were examined in lipid bilayers. Ivermectin (cis, 0.66-40 microM) activated six of eight native, four of four control-incubated and eleven of eleven FKBP12-'stripped' RyR channels. Midecamycin (cis, 10-30 microM) activated three of four single native channels, six of eight control-incubated channels and six of seven FKBP12 stripped channels. Activity declined when either drug was washed out. 3. Neither ivermectin nor midecamycin removed FKBP12 from RyRs. Western blots of terminal cisternae (TC), incubated for 15 min at 37 C with 40 microM ivermectin or midecamycin, showed normal amounts of FKBP12. In contrast, no FKBP12 was detected after incubation with 40 microM rapamycin. 4. Ivermectin reduced Ca2+ uptake by the SR Ca2+-Mg2+-ATPase. Ca2+ uptake by TC fell to approximately 40% in the presence of ivermectin (10 microM), both with and without 10 microM Ruthenium Red. Ca2+ uptake by longitudinal SR also fell to approximately 40% with 10 microM ivermectin. Midecamycin (10 microM) reduced Ca2+ uptake by TC vesicles to approximately 76% without Ruthenium Red and to approximately 90 % with Ruthenium Red. 5. The rate of rise of extravesicular [Ca2+] increased approximately 2-fold when 10 microM ivermectin was added to TC vesicles that had been partially loaded with Ca2+ and then Ca2+ uptake blocked by 200 nM thapsigargin. Ivermectin also potentiated caffeine-induced Ca2+ release to approximately 140% of control. These increases in Ca2+ release were not seen with midecamycin. 6. Ivermectin, but not midecamycin, reversibly reduced Ca2+ loading in four of six skinned rat extensor digitorum longus (EDL) fibres to approximately 90%, and reversibly increased submaximal caffeine-induced contraction in five of eight fibres by approximately 110% of control. Neither ivermectin nor midecamycin altered twitch or tetanic tension in intact EDL muscle fibres within 20 min of drug addition. 7. The results confirm the hypothesis that compounds with a macrocyclic lactone ring structure can directly activate RyRs. Unexpectedly, ivermectin also reduced Ca2+ uptake into the SR. These effects of ivermectin on SR Ca2+ handling may explain some effects of the macrolide drugs on mammals. PMID- 9852317 TI - Cell-type specific expression of ATP-sensitive potassium channels in the rat hippocampus. AB - 1. The distribution of ATP-sensitive K+ channels (KATP channels) was investigated in four cell types in hippocampal slices prepared from 10- to 13-day-old rats: CA1 pyramidal cells, interneurones of stratum radiatum in CA1, complex glial cells of the same area and granule cells of the dentate gyrus. The neuronal cell types were identified visually and characterized by the shapes and patterns of their action potentials and by neurobiotin labelling. 2. The patch-clamp technique was used to study the sensitivity of whole-cell currents to diazoxide (0.3 mM), a KATP channel opener, and to tolbutamide (0.5 mM) or glibenclamide (20 microM), two KATP channel inhibitors. The fraction of cells in which whole-cell currents were activated by diazoxide and inhibited by tolbutamide was 26% of pyramidal cells, 89 % of interneurones, 100% of glial cells and 89% of granule cells. The reversal potential of the diazoxide-induced current was at the K+ equilibrium potential and a similar current activated spontaneously when cells were dialysed with an ATP-free pipette solution. 3. Using the single-cell RT-PCR method, the presence of mRNA encoding KATP channel subunits (Kir6.1, Kir6.2, SUR1 and SUR2) was examined in CA1 pyramidal cells and interneurones. Subunit mRNA combinations that can result in functional KATP channels (Kir6.1 together with SUR1, Kir6.2 together with SUR1 or SUR2) were detected in only 17% of the pyramidal cells. On the other hand, KATP channels may be formed in 75% of the interneurones, mainly by the combination of Kir6.2 with SUR1 (58% of all interneurones). 4. The results of these combined analyses indicate that functional KATP channels are present in principal neurones, interneurones and glial cells of the rat hippocampus, but at highly different densities in the four cell types studied. PMID- 9852318 TI - A rise in intracellular Ca2+ underlies light adaptation in dogfish retinal 'on' bipolar cells. AB - 1. This investigation was to determine the extent to which retinal 'on' bipolar cells contribute to the adaptive changes that occur with light, which enable the rod visual system to operate over a wide range of ambient light intensities, and to elucidate the underlying adaptive mechanism. 2. Whole-cell voltage clamp recordings were obtained from bipolar cells in dark-adapted dogfish retinal slices. Current responses to brief flashes and steps of light were analysed. 'On' bipolar cell inward current light responses are mediated by a metabotropic glutamate receptor linked to the control of a cGMP cascade, with cGMP opening cation channels. Outward current responses to light of 'off' bipolar cells are mediated by the closure of ionotropic glutamate receptor channels. 3. When Ca2+ buffer was omitted from the patch pipette solution, 'on' bipolar cells rapidly desensitized to steps of light as dim as one rhodopsin molecule bleached per rod per second (1 Rh* s-1), whereas 'off' bipolar cells did not desensitize. Responses of 'on' bipolar cells to flashes in the presence of dim backgrounds recovered after a delay, but with diminished sensitivity, i.e. the cells adapted. 4. With the Ca2+ chelator BAPTA in the patch pipette solution, step responses of 'on' bipolar cells were sustained and flash responses following steps showed rapid recovery. Buffering Ca2+ in the patch pipette solution to 1 microM prevented desensitization, whereas 50 microM free Ca2+ reduced the 'on' bipolar cell flash responses, suppressed inward dark current and decreased input conductance. 5. We conclude that a major component of adaptation of the visual system is due to a reduction in gain at the rod-'on' bipolar cell synapse as a result of Ca2+ loading of the dendrites when their cGMP-gated cation channels open with light. PMID- 9852319 TI - Evidence that multiple P2X purinoceptors are functionally expressed in rat supraoptic neurones. AB - 1. The expression, distribution and function of P2X purinoceptors in the supraoptic nucleus (SON) were investigated by reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, and Ca2+-imaging and whole-cell patch-clamp techniques, respectively. 2. RT-PCR analysis of all seven known P2X receptor mRNAs in circular punches of the SON revealed that mRNAs for P2X2, P2X3, P2X4, P2X6 and P2X7 receptors were expressed in the SON, and mRNAs for P2X3, P2X4 and P2X7 were predominant. 3. In situ hybridization histochemistry for P2X3 and P2X4 receptor mRNAs showed that both mRNAs were expressed throughout the SON and in the paraventricular nucleus (PVN). 4. ATP caused an increase in [Ca2+]i in a dose-dependent manner with an ED50 of 1.7 x 10-5 M. The effects of ATP were mimicked by ATPgammaS and 2-methylthio ATP (2MeSATP), but not by AMP, adenosine, UTP or UDP. alphabeta-Methylene ATP (alphabetaMeATP) and ADP caused a small increase in [Ca2+]i in a subset of SON neurones. 5. The P2X7 agonist 2'- & 3'-O (4-benzoylbenzoyl)-ATP (BzATP) at 10-4 M increased [Ca2+]i, but the potency of BzATP was lower than that of ATP. In contrast, BzATP caused a more prominent [Ca2+]i increase than ATP in non-neuronal cells in the SON. 6. The effects of ATP were abolished by extracellular Ca2+ removal or by the P2 antagonist pyridoxal phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS), and inhibited by extracellular Na+ replacement or another P2 antagonist, suramin, but were unaffected by the P2X7 antagonist oxidized ATP, and the inhibitor of Ca2+-ATPase in intracellular Ca2+ stores cyclopiazonic acid. 7. Two patterns of desensitization were observed in the [Ca2+]i response to repeated applications of ATP: some neurones showed little or moderate desensitization, while others showed strong desensitization. 8. Whole-cell patch-clamp analysis showed that ATP induced cationic currents with marked inward rectification. The ATP-induced currents exhibited two patterns of desensitization similar to those observed in the [Ca2+]i response. 9. The results suggest that multiple P2X receptors, including P2X3, are functionally expressed in SON neurones, and that activation of these receptors induces cationic currents and Ca2+ entry. Such ionic and Ca2+ signalling mechanisms triggered by ATP may play an important role in the regulation of SON neurosecretory cells. PMID- 9852320 TI - Brainstem pathways responsible for oesophageal control of gastric motility and tone in the rat. AB - 1. Previous anatomical studies indicate that the nucleus of the solitary tract, pars centralis (NSTc) contains the neurones which receive vagal afferent input from the oesophagus. The purpose of the present study was to characterize the NSTc circuits in the medulla that may be responsible for oesophageal control of gastric motility. 2. Moderate balloon distension of the oesophagus of the rat (14 18 mmHg) provoked a significant reduction in gastric motility and tone recorded with strain gauges. This receptive relaxation effect was eliminated by bilateral lesions centred on the NSTc. 3. NSTc cells activated by oesophageal distension were labelled extracellularly and juxtacellularly with neurobiotin. NSTc neurones send axonal projections throughout the entire rostral-caudal extent of the dorsal motor nucleus of the vagus (DMN). These NSTc-DMN connections were confirmed by retrograde transport of neurobiotin from DMN to NSTc. NSTc neurones were observed with dendrites arborizing within the ependymal lining of the fourth ventricles. Thus, NSTc neurones may be in position to monitor blood-borne or ventricular agents and to alter the function of gastric-vago-vagal reflexes in response to these stimuli. 4. Neurophysiological recordings identified two subpopulations of DMN neurones which may be either activated or inhibited by oesophageal distension. Neurones excited by oesophageal distension were located mainly lateral and caudal in the DMN; neurones inhibited by oesophageal stimulation were located in medial and rostral DMN. 5. Our neurobiotin tracing results verified earlier studies showing that the NSTc projects to the intermediate reticular nucleus and the compact division of the nucleus ambiguus. Additionally, we found that the NSTc may be involved in reciprocal connections with the anterior, rostrolateral NST. 6. These results suggest that the gastric relaxation evoked by oesophageal distension is critically dependent on intact brainstem vago-vagal circuits. The NSTc, the recipient of oesophageal afferent projections from the vagus nerve, sends axons to the entire DMN, the source of parasympathetic control of the stomach. DMN neurones respond differentially to oesophageal distension, reinforcing the view that oesophageal afferents may provoke gastric relaxation by activating a vagal inhibitory pathway while simultaneously inhibiting a vagal excitatory pathway. PMID- 9852321 TI - Glucocorticoid modulation of Ca2+ homeostasis in human B lymphoblasts. AB - 1. We determined the effect of cortisol (200 nM for 48 h) on the intracellular Ca2+ concentration ([Ca2+]i) and parameters of Ca2+i signalling in 19 lymphoblastoid cell lines (LCLs). 2. Using the fluorescent dye fura-2, the basal [Ca2+]i in Ca2+-containing medium was 63.5 +/- 2.4 nM in vehicle (ethanol) treated LCLs and 55.7 +/- 2. 6 nM (mean +/- s.e.m.) in cortisol-treated LCLs. 3. Ca2+i signalling following platelet-activating factor (PAF, 100 nM) addition was enhanced by cortisol treatment, with LCLs having small PAF responses showing the largest percentage increase after cortisol treatment. Mean peak [Ca2+]i responses to PAF were enhanced 67.0% and 55.7% in Ca2+-free and Ca2+-containing medium, respectively. 4. The endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin (100 nM) caused a transient increase in [Ca2+]i in Ca2+-free medium in which the peak change was increased in cortisol-treated cells (98.5 +/- 5.8 vs. 79.8 +/- 4.5 nM). Peak changes in the freely exchangeable Ca2+ in response to 5 microM ionomycin were also enhanced in cortisol-treated cells (923.7 +/- 113.9 vs. 652.2 +/- 64.5 nM) and correlated to the PAF-evoked [Ca2+]i response. 5. Cortisol treated LCLs exposed to thapsigargin to empty intracellular Ca2+ stores (10 min treatment in Ca2+-free medium) and exposed to CaCl2 or MnCl2 had a greater rate of Ca2+ entry (18.6 +/- 1.8 vs. 13.8 +/- 1.5 nM s-1) and higher rate constant for Mn2+ entry (0.0345 +/- 0.0029 vs. 0. 0217 +/- 0.0020) than vehicle-treated cells. Peak [Ca2+]i in cells exposed to CaCl2 was also enhanced (869.4 +/- 114.7 vs. 562.6 +/- 61.7 nM). Parameters of divalent cation influx were highly correlated to the peak [Ca2+]i elicited by thapsigargin or ionomycin. 6. Inclusion of RU 486 (a glucocorticoid antagonist) with cortisol prevented the decrease in basal [Ca2+]i and stimulatory actions of cortisol on all Ca2+i parameters. RU 486 alone had no apparent effects on basal [Ca2+]i or Ca2+i signalling. 7. Based on data obtained over a wide range of responses (in the presence and/or absence of cortisol or RU 486), the results show that cortisol stimulation of glucocorticoid receptors decreases basal [Ca2+]i and enhances PAF-evoked [Ca2+]i signalling, most probably through its effects on intracellular Ca2+ stores. In turn, the extent of Ca2+ entry via store-operated plasma membrane Ca2+ channels is closely linked to the size of the Ca2+ stores. PMID- 9852322 TI - Divalent cation permeability and blockade of Ca2+-permeant non-selective cation channels in rat adrenal zona glomerulosa cells. AB - 1. The effects of the divalent cations Ca2+, Mg2+ and Ni2+ on unitary Na+ currents through receptor-regulated non-selective cation channels were studied in inside-out and cell-attached patches from rat adrenal zona glomerulosa cells. 2. External Ca2+ caused a concentration-dependent and voltage-independent inhibition of inward Na+ current, exhibiting an IC50 of 1.4 mM. The channel was also Ca2+ permeant and external Ca2+ shifted the reversal potential as expected for a channel exhibiting a constant Ca2+ : Na+ permeability ratio near to 4. 3. External and internal 2 mM Mg2+ caused voltage-dependent inhibition of inward and outward Na+ current, respectively. Modelling Mg2+ as an impermeant fast open channel blocker indicated that external Mg2+ blocked the pore at a single site exhibiting a zero voltage Kd of 5.1 mM for Mg2+ and located 19 % of the distance through the transmembrane electric field from the external surface. Internal Mg2+ blocked the pore at a second site exhibiting a Kd of 1.7 mM for Mg2+ and located 36% of the distance through the transmembrane electric field from the cytosolic surface. 4. External Ni2+ caused a voltage- and concentration-dependent slow blockade of inward Na+ current. Modelling Ni2+ as an impermeant slow open channel blocker indicated that Ni2+ blocked the pore at a single site exhibiting a Kd of 1.09 mM for Ni2+ and located 13.7% of the distance through the transmembrane electric field from the external surface. 5. External 2 mM Mg2+ increased the Kd for external Ni2+ binding to 1.27 mM, consistent with competition for a single binding site. Changing ionic strength did not substantially affect Ni2+ blockade indicating the absence of surface potential under physiological ionic conditions. 6. It is concluded that at least two divalent cation binding sites, separated by a high free energy barrier (the selectivity filter), are located in the pore and contribute to Ca2+ selectivity and permeability of the channel. PMID- 9852324 TI - Gating of skeletal and cardiac muscle sodium channels in mammalian cells. AB - 1. Sodium channel ionic current (INa) and gating current (Ig) were compared for rat skeletal (rSkM1) and human heart Na+ channels (hH1a) heterologously expressed in cultured mammalian cells at approximately 13 C before and after modification by site-3 toxins (Anthopleurin A and Anthopleurin B). 2. For hH1a Na+ channels there was a concordance between the half-points (V ) of the peak conductance voltage (G-V) relationship and the gating charge-voltage (Q-V) relationship with no significant difference in half-points. In contrast, the half-point of the Q-V relationship for rSkM1 Na+ channels was shifted to more negative potentials compared with its G-V relationship with a significant difference in the half points of -8 mV. 3. Site-3 toxins slowed the decay of INa in response to step depolarizations for both rSkM1 and hH1a Na+ channels. The half-point of the G-V relationship in rSkM1 Na+ channels was shifted by -8.0 mV while toxin modification of hH1a Na+ channels produced a smaller hyperpolarizing shift of the V by -3.7 mV. 4. Site-3 toxins reduced maximal gating charge (Qmax ) by 33% in rSkM1 and by 31% in hH1a, but produced only minor changes in the half-points and slope factors of their Q-V relationships. In contrast to measurements in control solutions, after modification by site-3 toxin the half-points of the G-V and the Q-V relationships for rSkM1 Na+ channels demonstrated a concordance similar to that for hH1a. 5. Qmax vs. Gmax for rSkM1 and hH1a Na+ channels exhibited linear relationships with almost identical slopes, as would be expected if the number of electronic charges (e-) per channel was comparable. 6. We conclude that the faster kinetics in rSkM1 channels compared with hH1a channels may arise from inherently faster rate transitions in skeletal muscle Na+ channels, and not from major differences in the voltage dependence of the channel transitions. PMID- 9852323 TI - Isoprenaline can activate the acetylcholine-induced K+ current in canine atrial myocytes via Gs-derived betagamma subunits. AB - 1. G protein betagamma subunits activate the acetylcholine-induced potassium current IK,ACh. There is no evidence of specificity at the level of the betagamma subunits. Therefore all G protein-coupled receptors in atrial myocytes should be able to activate IK,ACh. Paradoxically, it is often stated that isoprenaline does not activate IK,ACh. Rationales to explain this negative result include insufficient concentrations of Gs in the atrium or restricted access of Gs derived betagamma subunits to the IK,ACh channel. We took advantage of a non specific increase in Gs that results after infection with adenovirus. 2. Adenoviral infection unmasked a 1 microM isoprenaline-induced IK,ACh which was prevented by propranolol. Isoprenaline occasionally activated IK,ACh in uninfected and freshly dissociated atrial myocytes but the effect was larger and more consistent in infected myocytes. 3. Pertussis toxin pretreatment (100 ng ml 1 overnight) did not block the effect of isoprenaline. The effect of isoprenaline became persistent if cells were pretreated with cholera toxin (200 ng nl-1). 4. Signal transduction events distal to adenylyl cyclase were not involved in isoprenaline-induced IK,ACh. Forskolin (10 microM) did not activate IK,ACh. Inhibition of adenylyl cyclase with cytoplasmic application of 300 microM 2' deoxyadenosine 3'-monophosphate did not prevent the activation of IK,ACh by isoprenaline. 5. Cytoplasmic application of a betagamma binding peptide derived from the C terminus of beta-adrenergic receptor kinase 1 (50 microM) prevented the effect of isoprenaline on IK,ACh. The peptide did not prevent the stimulation of the L-type calcium current by isoprenaline. 6. The results indicate that beta adrenoceptors can activate IK,ACh in atrial myocytes through the release of betagamma subunits from Gs. PMID- 9852325 TI - High intracellular chloride delays the activation of the volume-sensitive chloride conductance in mouse L-fibroblasts. AB - 1. The relationship between cell volume and volume-sensitive Cl- conductance during hyposmotic cell swelling of patched cells and the effects of intracellular chloride on the conductance have been studied in mouse L-fibroblasts. To this end, swelling-activated current and cell volume were measured simultaneously in cells dialysed with low-Cl- (16 mM) or high-Cl- (130 mM) solutions using the whole-cell patch-clamp technique and videomicroscopy. 2. The increase in cell volume of patched cells and the volume-sensitive conductance saturated during a 4 5 min exposure to mildly hyposmotic solutions (15-20 % less than isosmotic). The swelling of patched cells varied considerably and was greater than the swelling of intact cells. No correlation between the maximal values of the volume sensitive conductance and the maximal volumes of swollen cells was evident for cells dialysed with the low-Cl- solutions. 3. The amplitude of the volume sensitive conductance decreased with a reduction in either extracellular or intracellular Cl- concentration; the size of the maximal conductance was not modulated by intracellular Cl- ions. 4. The activation of the volume-sensitive conductance was slower in high-Cl- cells than in low-Cl- cells whether it was induced by hypotonic cell swelling or by cell inflation; in low-Cl- cells the conductance saturated before the cell volume had reached its maximal value. 5. It is concluded that in patched cells an increase in cell volume triggers activation of the volume-sensitive Cl- conductance but does not determine its amplitude and that the rate of activation of the conductance is affected by the intracellular Cl- concentration. PMID- 9852326 TI - Sr2+-dependent asynchronous evoked transmission at rat striatal inhibitory synapses in vitro. AB - 1. At striatal inhibitory synapses in cell culture, replacement of extracellular Ca2+ with Sr2+ desynchronized inhibitory postynaptic currents (IPSCs), reducing their peak amplitude and producing a succession of late, asynchronous synaptic events (late release). In the averaged IPSC waveform this resulted in an increase in both the fast and the slow decay time constant as well as in the time to peak. 2. Rapid removal of extracellular Sr2+ during late release was without effect on the time course of the averaged IPSC. Thus, late release is not dependent on continuous Sr2+ influx, but must be related to the way in which Sr2+, as opposed to Ca2+, interacts with constituents of the intracellular space. 3. After application of the membrane-permeant acetoxymethyl ester (AM) form of the Ca2+ chelator BAPTA, Sr2+-induced late release was greatly reduced and the kinetics of the Sr2+-dependent IPSC approached those of the Ca2+-dependent response. EGTA AM had a similar but less pronounced effect. 4. Using rapid solution exchange, we stimulated synapses first in Sr2+- or Ca2+- and 100-300 ms afterwards in Ca2+ containing solution. Paired-pulse facilitation of late release was the same whether the conditioning pulse induced a presynaptic influx of Sr2+ or of Ca2+. 5. It is concluded that Sr2+-mediated asynchrony is probably due to a less efficient intraterminal buffering of Sr2+ as opposed to Ca2+, allowing for Sr2+ ions to activate release in an area less confined to the immediate vicinity of the presynaptic Ca2+ channel. This hypothesis explains both the action of endogenous buffers and the apparent lack of specific facilitatory interaction between Ca2+-mediated and Sr2+-induced late release. PMID- 9852327 TI - Spontaneous and neurally activated depolarizations in smooth muscle cells of the guinea-pig urethra. AB - 1. Membrane potential recordings were made from longitudinal smooth muscle cells of the guinea-pig urethra using conventional microelectrode techniques. 2. Smooth muscle cells of the urethra developed spontaneous transient depolarizations (STDs) and slow waves. Single unit STDs had amplitudes of approximately 5 mV and slow waves seemed to occur as amplitude multiples of single unit STDs. 3. STDs and slow waves were abolished by niflumic acid or low chloride solution and also by cyclopiazonic acid (CPA), BAPTA or high concentrations of caffeine. Lower concentrations of caffeine abolished slow waves but not STDs. Nifedipine inhibited slow waves but not STDs. 4. When stochastic properties of STDs were examined, it was found that the intervals between occurrences were not well modelled by Poisson statistics, instead the STDs appeared to be clustered. 5. Transmural stimulation evoked excitatory junctional potentials (EJPs) and triggered slow waves which were abolished by either alpha,beta-methylene-ATP or tetrodotoxin. Evoked slow waves were also abolished by caffeine, co-application of caffeine and ryanodine or by CPA which left EJPs unaffected. 6. In conclusion, smooth muscle cells of urethra exhibit STDs which are clustered rather than random events, and are the result of spontaneous Ca2+ release from intracellular stores and subsequent activation of Ca2+-activated chloride channels. STDs sum to activate L-type Ca2+ channels which contribute to the sustained phase of slow waves. Stimulation of purinoceptors by neurally released ATP initiates EJPs and also causes the release of Ca2+ from intracellular stores to evoke slow waves. PMID- 9852328 TI - Interference of H2O2 with stimulus-secretion coupling in mouse pancreatic beta cells. AB - 1. We have reported previously that in mouse pancreatic beta-cells H2O2 hyperpolarizes the membrane and increases the ATP-sensitive K+ current recorded in the perforated patch configuration of the patch-clamp technique. The present study was undertaken to elucidate the underlying mechanisms. 2. The intracellular ATP concentration measured by chemoluminescence was reduced by H2O2. The ADP concentration increased in parallel during the first 10 min, resulting in a pronounced decrease in the ATP/ADP ratio. 3. Consistent with these results, glucose-stimulated insulin secretion from isolated islets was inhibited by H2O2. 4. Membrane hyperpolarization measured with intracellular microelectrodes in intact islets and inhibition of insulin secretion were counteracted by tolbutamide, indicating that the channels are still responsive to inhibitors and that the ATP concentration is not too low to trigger exocytosis. However, the sensitivity of the beta-cells to tolbutamide was reduced after treatment with H2O2. 5. H2O2 increased the intracellular Ca2+ activity ([Ca2+]i) in a biphasic manner. A first transient rise in [Ca2+]i due to mobilization of Ca2+ from intracellular stores was followed by a sustained increase, which was at least partly dependent on Ca2+ influx. The first phase seems to reflect Ca2+ mobilization from mitochondria. 6. Our results demonstrate that H2O2 interferes with glucose metabolism, which influences the membrane potential and ATP sensitive K+ current via the intracellular concentration of ATP. These events finally lead to an inhibition of insulin secretion despite an increase in [Ca2+]i. PMID- 9852329 TI - Chronic hypoxia enhances the secretory response of rat phaeochromocytoma cells to acute hypoxia. AB - 1. Amperometric recordings were made from individual phaeochromocytoma (PC12) cells using carbon fibre microelectrodes to investigate the effects of chronic hypoxia (10% O2) on the secretory responses evoked by acute hypoxia. 2. Exposure to chronic hypoxia for 21-26 h increased the frequency of exocytotic events evoked in response to acute hypoxia (PO2 ca 10-60 mmHg). 3. Chronic hypoxia increased the value of Q1/3, determined by the integration of amperometric events, indicating an increase in quantal size: this reflects either an increase in vesicular dimensions or vesicular catecholamine concentration. 4. Exocytotic frequency evoked by bath application of tetraethylammonium (1-10 mM) was significantly enhanced following chronic hypoxia. 5. In both control and chronically hypoxic PC12 cells, exocytosis in response to acute hypoxia was completely abolished in Ca2+-free solutions. Cd2+ (200 microM) completely inhibited exocytosis from control cells, but left a significant residual release in chronically hypoxic PC12 cells. 6. The Cd2+-resistant release evoked by acute hypoxia in chronically hypoxic PC12 cells was inhibited by inorganic ions (0.01 10 mM) in a potency order of La3+ > Gd3+ > Zn2+. Ni2+ (10 mM) was without effect. 7. Our results suggest that chronic hypoxia enhances the secretory response of PC12 cells in part by increasing the depolarization mediated by an oxygen sensitive K+ channel. In addition, acute hypoxia activates a Cd2+-resistant Ca2+ influx pathway in chronically hypoxic PC12 cells. PMID- 9852330 TI - Resetting and postnatal maturation of oxygen chemosensitivity in rat carotid chemoreceptor cells. AB - 1. Carotid chemoreceptor sensitivity is minimal immediately after birth and increases with postnatal age. In the present study we have investigated the peri- and postnatal developmental time course of [Ca2+]i responses to hypoxia in clusters of type I cells isolated from near-term fetal rats and rats that were 1, 3, 7, 11, 14 and 21 days old, using the Ca2+-sensitive fluoroprobe fura-2. 2. In type I cells from all age groups a graded increase in [Ca2+]i occurred in response to lowering the PO2 from 150 mmHg to 70, 35, 14, 7, 2 and 0 mmHg. The graded [Ca2+]i response to hypoxia was hyperbolic at all ages. 3. Type I cells from rats near-term fetal to 1 day old exhibited small [Ca2+]i responses, mainly to the most severe levels of hypoxia. After day 1, an increase in the [Ca2+]i responses to submaximal hypoxia stimulation resulted in a rightward shift in the O2 response curve. Using the Delta[Ca2+]i between 35 and 2 mmHg PO2 as an index of O2 sensitivity, type I cell O2 sensitivity increased approximately 4- to 5 fold between near-term fetal to 1 day old and 11 to 14 days of age. 4. Exposure to elevated extracellular potassium (10, 20 and 40 mM K+) caused a dose-dependent [Ca2+]i rise in type I cells from all age groups. There were no age-related changes in [Ca2+]i responses to any level of K+ between near-term fetal and 21 days. 5. We conclude that the maximal type I cell [Ca2+]i response to anoxia, as well as the sensitivity to submaximal hypoxic stimulation, of rats aged from near term fetal to 21 days depends on the level of postnatal maturity. The lack of an age-related increase in the [Ca2+]i response to elevated K+ during the timeframe of maximal development of O2 sensitivity suggests that resetting involves maturation of O2 sensing, rather than non-specific developmental changes in the [Ca2+]i rise resulting from depolarization. PMID- 9852331 TI - Endothelium-dependent hyperpolarization and intercellular electrical coupling in guinea-pig mesenteric arterioles. AB - 1. Using the conventional whole-cell clamp method, the electrical responses of individual smooth muscle and endothelial cells to acetylcholine (ACh) were observed in multicellular preparations where the two types of cells remained in close apposition. 2. In both types of cells, ACh induced similar hyperpolarizing responses which, when recorded in current clamp mode, had two phases (an initial fast and a second slower phase). 3. After blocking gap junctions, including myoendothelial junctions, with 18beta-glycyrrhetinic acid, ACh induced an outward current with two phases in voltage-clamped endothelial cells. The outward current appeared around -90 mV and increased linearly with the membrane depolarization. 4. In smooth muscle cells, ACh failed to induce a membrane current after gap junctions had been blocked with 18beta-glycyrrhetinic acid. The inhibition of ACh induced response by 18beta-glycyrrhetinic acid was observed using either sharp or patch electrodes. 5. Nominally Ca2+-free solution reduced the initial phase and abolished the second phase of ACh-induced responses of endothelial cells. Both phases were also reduced by charybdotoxin (CTX). 6. Our results indicate that in guinea-pig mesenteric arterioles, ACh hyperpolarizes endothelial cells by activating Ca2+-activated K+ channels which are sensitive to CTX. On the other hand, hyperpolarizing responses detected in smooth muscle cells seem to originate in endothelial cells and conduct to the muscle layer via myoendothelial gap junctions. PMID- 9852332 TI - Identification of rhythmically active cells in guinea-pig stomach. AB - 1. When intracellular recordings were made from the antral region of guinea-pig stomach, cells with different patterns of electrical activity were detected. 2. One group of cells, slow-wave cells, generated slow waves which consisted of initial and secondary components. When filled with either Lucifer Yellow or neurobiotin, the cells identified as smooth muscle cells lying in the circular muscle layer. 3. A second group of cells, driving cells, generated large, rapidly rising, potential changes, driving potentials. They had small cell bodies with several processes. With neurobiotin, a network of cells was visualized that resembled c-kit positive interstitial cells of the myenteric region. 4. A third group of cells generated sequences of potential changes which resembled driving potentials but had smaller amplitudes and slow rates of rise. These cells resembled smooth muscle cells lying in the longitudinal muscle layer. 5. When simultaneous recordings were made from the driving and slow-wave cells, driving potentials and slow waves occurred synchronously. Current injections indicated that both cell types were part of a common electrical syncytium. 6. The initial component of slow waves persisted in low concentrations of caffeine, but the secondary component was abolished; higher concentrations shortened the duration of the residual initial component. Driving potentials continued in the presence of low concentrations of caffeine; moderate concentrations of caffeine shortened their duration. 7. Hence three different types of cells were distinguished on the basis of their electrical activity, their responses to caffeine and their structure. These were smooth muscle cells, lying in the longitudinal and circular layers, and interstitial cells in the myenteric region. The observations suggest that interstitial cells initiate slow waves. PMID- 9852333 TI - Calcium channels coupled to neurotransmitter release at neonatal rat neuromuscular junctions. AB - 1. The effects of different calcium channel blockers (omega-agatoxin IVA (omega Aga IVA), omega-conotoxin GVIA (omega-CgTx GVIA) and dihydropyridines) were tested on spontaneous and evoked transmitter release at embryonic and newborn rat neuromuscular junctions (NMJs). 2. The nerve-evoked transmitter release quantal content (m) was strongly reduced by the P/Q-type voltage-dependent calcium channel (VDCC) blocker omega-Aga IVA (100 nM) at newly formed endplates of embryos and 0- to 11-day-old rats, in agreement with the effect of this blocker on transmitter release at mature and reinnervating muscles. 3. omega-CgTx GVIA (1 5 microM), the N-type VDCC blocker, also caused a significant reduction in m at newly formed NMJs early in development (embryos and 0- to 4-day-old rats), while it was ineffective in more mature animals (5- to 11-day-old rats). 4. L-type channel blockers, nitrendipine (1 microM) and nifedipine (1 microM), did not significantly affect neurally evoked release at developing NMJs. However, nifedipine (10 microM) was able to increase m significantly at 0- to 4-day-old rat NMJs. 5. At developing NMJs, K+-evoked transmitter release was dependent on Ca2+ entry through VDCCs of the P/Q-type family (100 nM omega-Aga IVA reduced 70 % of the K+-evoked miniature endplate potential frequency). N- and L-type VDCC blockers did not affect this type of release. 6. We conclude that at rat neuromuscular junctions the presynaptic calcium channel types involved in transmitter release undergo developmental changes during the early postnatal period. PMID- 9852334 TI - Binocular phase interactions in area 21a of the cat. AB - 1. Binocular interactions related to retinal disparity were investigated in single neurons in area 21a of extrastriate cortex in the anaesthetized cat using sinusoidal luminance gratings. 2. The responses of approximately two-thirds of neurons were profoundly modulated by a relative phase difference between identical drifting gratings presented to each eye. This modulation included both facilitatory and inhibitory interocular interactions. The selectivity for binocular disparity was about twice as sharp as the selectivity for monocular spatial position. 3. Significant phase modulation was retained in many neurons at interocular orientation differences exceeding 45 deg. The response suppression associated with stimulation at a phase shift 180 deg from the optimum was stronger than the response suppression to an interocular orientation difference of 90 deg. 4. The proportion of phase modulated neurons and the potency of modulation in area 21a neurons exceed that reported for phase-selective complex cells in area 17. Neurons in area 21a show sharp disparity tuning that is relatively insensitive to changes in orientation and monocular position, which suggests that this extrastriate region has a role in stereoscopic depth perception. PMID- 9852335 TI - Topographical organization of projections to cat motor cortex from nucleus interpositus anterior and forelimb skin. AB - 1. The activation of the motor cortex from focal electrical stimulation of sites in the forelimb area of cerebellar nucleus interpositus anterior (NIA) was investigated in barbiturate-anaesthetized cats. Using a microelectrode, nuclear sites were identified by the cutaneous climbing fibre receptive fields of their afferent Purkinje cells. These cutaneous receptive fields can be identified by positive field potentials reflecting inhibition from Purkinje cells activated on natural stimulation of the skin. Thereafter, the sites were microstimulated and the evoked responses were systematically recorded over the cortical surface with a ball-tipped electrode. The topographical organization in the motor cortex of responses evoked by electrical stimulation of the forelimb skin was also analysed. 2. Generally, sites in the forelimb area of NIA projected to the lateral part of the anterior sigmoid gyrus (ASG). Sites in the hindlimb area of NIA also projected to lateral ASG and in addition to a more medial region. Sites in the face area of NIA, however, projected mainly to the middle part of the posterior sigmoid gyrus (PSG). 3. For sites in the forelimb area of NIA, the topographical organization and strength of the projections varied specifically with the cutaneous climbing fibre receptive field of the site. The largest cortical responses were evoked from sites with receptive fields on the distal or ventral skin of the forelimb. 4. Microelectrode recordings in the depth of the motor cortex revealed that responses evoked by cerebellar nuclear stimulation were due to an excitatory process in layer III. 5. Short latency surface responses evoked from the forelimb skin were found in the caudolateral part of the motor cortex. At gradually longer latencies, responses appeared in sequentially more rostromedial parts of the motor cortex. Since the responses displayed several temporal peaks that appeared in specific cortical regions for different areas of the forelimb skin, several somatotopic maps were seen. 6. The cerebellar and cutaneous projections activated mainly different cortical regions and had topographical organizations that apparently were constant between animals. Their patterns of activation may constitute a frame of reference for investigations of the functional organization of the motor cortex. PMID- 9852336 TI - Neurotransmitters and neuromodulators controlling the hypoxic respiratory response in anaesthetized cats. AB - 1. The contributions of neurotransmitters and neuromodulators to the responses of the respiratory network to acute hypoxia were analysed in anaesthetized cats. 2. Samples of extracellular fluid were collected at 1-1.5 min time intervals by microdialysis in the medullary region of ventral respiratory group neurones and analysed for their content of glutamate, gamma-aminobutyric acid (GABA), serotonin and adenosine by high performance liquid chromatography. Phrenic nerve activity was correlated with these measurements. 3. Levels of glutamate and GABA increased transiently during early periods of hypoxia, coinciding with augmented phrenic nerve activity and then fell below control during central apnoea. Serotonin and adenosine increased slowly and steadily with onset of hypoxic depression of phrenic nerve activity. 4. The possibility that serotonin contributes to hypoxic respiratory depression was tested by microinjecting the 5 HT-1A receptor agonist 8-OH-DPAT into the medullary region that is important for rhythmogenesis. Hypoxic activation of respiratory neurones and phrenic nerve activity were suppressed. Microinjections of NAN-190, a 5-HT-1A receptor blocker, enhanced hypoxic augmentation resulting in apneustic prolongation of inspiratory bursts. 5. The results reveal a temporal sequence in the release of neurotransmitters and neuromodulators and suggest a specific role for each of them in the sequential development of hypoxic respiratory disturbances. PMID- 9852337 TI - Gastrectomy induces impaired insulin and glucagon secretion: evidence for a gastro-insular axis in mice. AB - 1. Mice were subjected to gastrectomy (GX) or food deprivation (24 h). The release of insulin and glucagon in response to different secretagogues was monitored in vivo and in isolated islets 3-4 weeks after surgery. 2. GX animals responded to glucose with an impaired glucose tolerance and a poor increase in plasma insulin. Islets from GX or food-deprived mice displayed impaired insulin release to high glucose and enhanced glucagon release at low glucose. 3. After GX the insulinogenic index, Delta insulin (microU ml-1)/Delta glucose (mg ml-1), was suppressed by 65% after oral glucose and by 59% after i.v. glucose. The integrated insulin response after oral glucose was reduced by 90% in GX mice. After i.v. glucose the reduction was 67%. 4. Carbachol-induced insulin release in vivo was reduced after food deprivation and exaggerated after GX. Carbachol stimulated glucagon secretion was suppressed after GX and after food deprivation. A similar pattern was found in vitro. 5. Cyclic AMP activation (by the phosphodiesterase inhibitor isobutylmethylxanthine or the adenylate cyclase stimulator forskolin) induced a greater insulin response in GX or food-deprived mice than in sham-operated, fed mice. A similar pattern was found in vitro. The glucagon response was enhanced in vitro but not in vivo. 6. Crude extracts of rat oxyntic mucosa enhanced basal as well as glucose-induced insulin release from isolated islets, whereas glucagon release was markedly inhibited. The effects were dose dependent, the inhibition of glucagon release being achieved at lower concentrations than the potentiation of glucose-induced insulin release. The active principle was inactivated by incubation with trypsin or leucine aminopeptidase. 7. The data suggest that a circulating agent, probably a peptide, from gastric oxyntic mucosa stimulates glucose-induced insulin secretion. It also suppresses glucagon secretion. The GX-evoked impairment of the insulin (and glucagon) response to glucose is partly compensated for by an enhanced insulin response to cholinergic and/or cyclic AMP activation. PMID- 9852338 TI - Excitability changes in sacral afferents innervating the urethra, perineum and hindlimb skin of the cat during micturition. AB - 1. Excitability changes in afferents innervating the urethra, perineum and hindlimb were measured in decerebrated cats during micturition and in response to stimulation of lumbosacral afferents. Increases in excitability were interpreted as primary afferent depolarization (PAD) and decreases as primary afferent hyperpolarization. 2. Excitability increases were observed in 11 of 19 urethral pudendal afferents during micturition. Four of these 11 afferents showed an excitability increase during voiding. Seven of these showed a biphasic change with a decrease in excitability when sphincter activity resumed at the end of the void. Three of 19 afferents showed an excitability decrease during micturition and no change was detected in five afferents. 3. During micturition, the peak amplitude of urethral afferent-evoked excitatory postsynaptic potentials in seven of eight sphincter motoneurones was diminished to a mean of 36% of control values. 4. Eighty per cent of hindlimb cutaneous afferents and 50% of dorsal penile/clitoral and superficial perineal nerve afferents in the sacral cord showed increased excitability during voiding. No excitability increases were measured in 13 hindlimb cutaneous fibres examined in the lumbar segments. 5. PAD was observed in sacral urethral, perineal and hindlimb cutaneous afferents in response to electrical stimulation of other perineal, urethral, hindlimb cutaneous and group II muscle afferents. 6. It is concluded that control of transmission from urethral afferents by the micturition circuitry is different to that by sensory transmission from hindlimb and perineal regions during micturition. We hypothesize that more than one population of sacral PAD-mediating interneurones is involved. PMID- 9852339 TI - Perceptual distortions of the human body image produced by local anaesthesia, pain and cutaneous stimulation. AB - 1. Knowledge of the size and orientation of the hand is essential if it is to be moved accurately in space. We used two psychophysical methods to determine whether the perceived size of a body part changes when its sensory input is changed: first, the selection of scaled drawings which matched the apparent size of a body part, and second, a motor task in which the subject drew the body part to depict its perceived size. 2. Complete anaesthesia of the thumb (with a digital nerve block) significantly increased its perceived size by 60-70% when assessed with both psychophysical methods. During this anaesthesia, the perceived size of the adjacent index finger or digits on the contralateral side was unaltered. However, the size of the unanaesthetized lips increased (by approximately 50%). 3. Marked sensory loss for the lips (produced by topical anaesthetics) significantly increased their perceived size when assessed with both methods of measurement. There was a small increase in apparent size of the thumb. 4. To determine whether changes in perceived size could also be produced by an elevation of peripheral inputs, innocuous electrical stimulation of the digital nerves and also painful cooling of the digit were used. Both procedures produced small but significant increases in perceived size of the stimulated part. 5. The results highlight lability in the perceived size of parts of the body and how this affects motor output. The data may reveal perceptual consequences of acute changes in central somatosensory maps, changes which are known to occur with deafferentation. PMID- 9852340 TI - Perspectives on pituitary adenylate cyclase activating polypeptide (PACAP) in the neuroendocrine, endocrine, and nervous systems. AB - PACAP is a pleiotropic neuropeptide that belongs to the secretin/glucagon/VIP family. PACAP functions as a hypothalamic hormone, neurotransmitter, neuromodulator, vasodilator, and neurotrophic factor. Its structure has been remarkably conserved during evolution. The PACAP receptor is G protein-coupled with seven transmembrane domains and also belongs to the VIP receptor family. PACAP, but not VIP, binds to PAC1-R, whereas PACAP and VIP bind to VPAC1-R and VPAC2-R with a similar affinity. Despite the sizable homology of the structures of PACAP and VIP and their receptors, the distribution of these peptides and receptors is quite different. At least eight subtypes of PACAP specific, or PAC1 R, result from alternate splicing. Each subtype is coupled with specific signaling pathways, and its expression is tissue or cell specific. Although PACAP fulfills most requirements for a physiological hypothalamic hypophysiotropic hormone, it does not consistently stimulate secretion of the adenohypophysial hormones, except for stimulation of IL-6 release from the FS cells of the pituitary. The major regulatory role of PACAP in pituitary cells appears to be the regulation of gene expression of pituitary hormones and/or regulatory proteins that control growth and differentiation of the pituitary glandular cells. These effects appear to be exhibited directly and indirectly through a paracrine or autocrine action. Although PACAP stimulates the release of AVP, the physiological role of neurohypophysial PACAP remains unknown. One important action of PACAP in the endocrine system is its role as a potent secretagogue for adrenaline from the adrenal medulla through activation of TH. PACAP also stimulates the release of insulin and increases [Ca2+]i from pancreatic beta cells at an extremely small concentration. The stage-specific expression of PACAP in testicular germ cells during spermatogenesis suggests its regulatory role in the maturation of germ cells. In the ovary, PACAP is transiently expressed in the granulosa cells of the preovulatory follicles and appears to be involved in the LH-induced cellular events in the ovary, including prevention of follicular apoptosis. In the central nervous system, PACAP acts as a neurotransmitter or neuromodulator, which has been supported by IHC and electrophysiological methods. More important, PACAP is a neurotrophic factor that may play an important role during the development of the brain. In the adult brain, PACAP appears to function as a neuroprotective factor that attenuates the neuronal damage resulting from various insults. PMID- 9852342 TI - An increase in the threshold of citric acid-induced cough during chest wall vibration in healthy humans. AB - This study tested the hypothesis that the afferent input from the respiratory muscles may be involved in the neural mechanisms inducing cough responses. Coughing was evoked in conscious healthy humans by the inhalation of citric acid aerosol of several concentrations either during or not during chest wall vibration (100 Hz) at the right second intercostal space or during vibration of the right thigh. The mean threshold citric acid concentration to induce coughing was significantly higher during chest wall vibration (geometric mean, 131.8 mg/ml) than without vibration (75.9 mg/ml). Vibration after topical anesthesia of the chest wall skin did not significantly change the threshold concentration of citric acid. The threshold citric acid concentration during vibration of the right thigh did not significantly differ from that without vibration. We concluded that inputs from the chest wall afferent, presumably from the intercostal muscle or costovertebral joint, may have an inhibitory effect on the initiation of coughing at the higher neural structure in conscious humans. PMID- 9852341 TI - Plasma levels of 5-HT and 5-HIAA increased after intestinal ischemia/reperfusion in rats. AB - Intestinal ischemia/reperfusion (I/R) causes serious systemic injury, mainly from a variety of bioactive substances released from the injured intestine. To assess the possible roles of serotonin (5-hydroxytryptamine, 5-HT), a bioactive amine mainly stored in the intestine, in I/R injury, we assayed the levels of tryptophan, 5-HT, and 5-hydroxyindole acetic acid (5-HIAA) in the blood and intestine in a rat I/R model. Plasma 5-HT increased significantly over time after reperfusion; the plateau level was obtained 4 h after reperfusion and was associated with an increase in 5-HIAA. Plasma tryptophan levels declined gradually after reperfusion. The ratio of 5-HIAA/5-HT was significantly higher in I/R rats than in control rats, suggesting that elevated 5-HT was quickly metabolized in the systemic circulation. In the intestine, 5-HT decreased dramatically, whereas tryptophan increased. This phenomenon was prominent in the severely damaged intestine. These findings suggest that the injured intestine released large amounts of 5-HT, whereas its synthesis in the injured intestine was suppressed. An increase in 5-HT in the circulation may be related to various circulatory disturbances observed in humans after intestinal ischemia. PMID- 9852343 TI - Unloaded skeletal muscle O2 uptake decreases with decreased venous PO2 at high frequency stimulation. AB - The aim of the present study was to clarify the effects of O2 diffusion limitation resulting from hypoxic interventions on O2 uptake (V.O2) in unloaded (that is, near-zero initial force) and loaded skeletal muscle in a high-frequency stimulation. We measured V.O2, muscle venous PO2 (PvO2) and initial force in gastrocnemius-plantaris muscle in situ of anesthetized dogs: (1) during hypoxic hypoxia at 1 Hz tetanic stimulation, and (2) during hypoxia induced by the perfusion with high O2-affinity erythrocytes (having a low value of PO2 at 50% saturation of hemoglobin (P50)) at 4 Hz twitch stimulation. Averaged unloaded V.O2 during normoxia was 10.2 ml.min-1.100 g-1 at averaged blood flow of 74 ml.min-1.100 g-1 (n = 6). Hypoxic hypoxia of a decreased O2 delivery (arterial O2 concentration x flow) significantly decreased both unloaded and loaded V.O2 with a decrease in PvO2 (p<0.05). The unloaded V.O2 was reduced to 8.5 ml.min-1.100 g 1. Low P50-hypoxia decreased V.O2 at high and low initial force conditions with a decrease in PvO2 (p<0.05) at the same O2 delivery. If these decreases in V.O2 correspond with a decrease in V.O2 at zero initial force (unloaded V.O2), the unloaded V.O2 value is calculated to be 7.57 ml.min-1.100 g-1 from V. O2-initial force data. Despite the different conditions of O2 delivery, the unloaded V.O2 decreased by both hypoxia showed similar values. Thus the decreased unloaded V.O2 does not seem to be derived from only the limited O2 delivery. Some other factors such as the limitation of O2 diffusion may contribute to the decreased V.O2. PMID- 9852345 TI - Endurance exercise training increases peripheral vascular response in human fingers. AB - The purpose of this study was to clarify whether peripheral vascular response to alteration of transmural pressure is changed by endurance exercise training. The healthy male subjects (training group; n = 6) performed endurance exercise training that consisted of cycle ergometer exercise 5 d.week-1 and 30 min.d-1 for a period of 8 weeks. Changes in the peripheral vascular response to alteration of transmural pressure in the human finger were measured by a differential digital photoplethysmogram (DeltaDPG) and blood pressure during passive movement of the arm to different vertical hand positions relative to heart level. Following 8 weeks of endurance training, percent changes in DeltaDPG from heart level in the training group increased significantly (mean +/- SD, -48.1 +/- 7. 3 to -58.7 +/- 9.3% at the lowered position, 46.1 +/- 13.4 to 84.6 +/- 8.8% at the elevated position, p<0.05). Similarly, the arterial compliance index, which was calculated from DeltaDPG-P wave amplitude and arterial pulse pressure, also significantly changed in the training group over the 8 weeks (5.6 +/- 1.3 to 2.7 +/- 1.6 mV. V 1.s-1.mmHg-1 at the lowered position, 30.0 +/- 12.4 to 54.4 +/- 18. 9 mV.V-1.s 1.mmHg-1 at the elevated position ). Maximal oxygen uptake (V.O2 max) was significantly increased in the training group. On the other hand, the control group (n = 6) showed no significant changes in all parameters for 8 weeks. Therefore these results suggest that endurance exercise training induces an increase in peripheral vascular response to alteration of transmural pressure in the human finger. PMID- 9852344 TI - Thermogenesis and fatty acid composition of brown adipose tissue in rats rendered hyperthyroid and hypothyroid-with special reference to docosahexaenoic acid. AB - The effects of hyperthyroidism and hypothyroidism on brown adipose tissue (BAT) thermogenesis and phospholipid fatty acid composition were investigated in rats. Chronic triiodothyronine (T3) treatment (hyperthyroidism) increased the interscapular BAT pad weight, its triacylglycerol content, and its DNA content. It did not affect basal and noradrenaline-stimulated in vitro oxygen consumption of BAT expressed per microg DNA, although it significantly increased the oxygen consumption of the whole BAT pad. T3 treatment had little effect on phospholipid content and phospholipid fatty acid composition. In contrast, chronic methimazole treatment (hypothyroidism) decreased the BAT pad weight and the triacylglycerol content, but did not significantly change the DNA content in comparison with the control. It significantly decreased the noradrenaline-stimulated BAT oxygen consumption expressed per microg DNA and per BAT pad, but did not change the basal oxygen consumption. Methimazole treatment significantly affected phospholipid content and phospholipid fatty acid composition. Among the major fatty acids of BAT, it decreased docosahexaenoic acid (DHA), arachidonic acid, palmitic acid, palmitoleic acid, and oleic acid, and it increased linoleic acid, stearic acid, and eicosapentaenoic acid. A regression analysis revealed a positive relationship between in vitro respiration and DHA levels in phospholipids (r = 0.404, p<0.05). These results suggest that thyroid hormones have trophic action on BAT and are necessary for BAT thermogenic activity. This study also suggests that DHA is involved in the regulation of BAT thermogenic activity, as we previously indicated. PMID- 9852346 TI - Effects of intracoronary caffeine on left ventricular mechanoenergetics in Ca2+ overload failing rat hearts. AB - How different the effects of caffeine on cardiac mechanoenergetics in failing hearts are from those of normal hearts remains to be fully elucidated. First we successfully instituted a new experimental model of acute mild heart failure in the rat by 0.005 mM Ca2+ Tyrode perfusion. These failing hearts neither decreased left ventricular end-systolic pressure nor increased left ventricular end diastolic pressure, indicating unchanged left ventricular mechanics. However, their myocardial mitochondrial respiratory function examined by respiratory control index (RCI) and oxygen consumption rate in state III (State III O2) was significantly depressed compared with normal hearts. From these results, we judged that this Ca2+ protocol could make mild Ca2+ overload acute failing hearts and that this model would be appropriate for comparing the effects of caffeine on cardiac mechanoenergetics between normal hearts and these failing hearts. We investigated the effects of caffeine on cardiac mechanoenergetics above a concentration of 0.05 mM that corresponds to the maximum blood concentration after a healthy human subject drinks a cup of coffee or tea. We obtained results indicating that caffeine depressed left ventricular systolic and diastolic functions and decreased a measure of total mechanical energy per beat in terms of systolic pressure-volume area (PVA) more severely in these failing hearts at concentrations (20-fold higher than the concentration in a cup of coffee) lower than those in normal hearts. This result implies that these acute failing hearts are Ca2+ overloaded. PMID- 9852347 TI - Evidence for the essential role of myosin subfragment-2 in the ATP-dependent actin-myosin sliding in muscle contraction. AB - The role of myosin subfragment-2 (myosin S-2) in muscle contraction was studied by using an in vitro motility assay system in which the ATP-dependent sliding between myosin-coated polystyrene beads and actin filament arrays (actin cables) of giant algal cells were recorded under constant external loads provided with a centrifuge microscope. With antibody to myosin S-2 below 0.3 mg/ml, the maximum "isometric" force generated by myosin molecules on the bead decreased markedly, but the unloaded bead-sliding velocity along actin cables did not change appreciably, indicating a decrease in the number of myosin molecules interacting with actin cables. The antibody at 0.3-1.5 mg/ml decreased not only the maximum isometric force, but also the unloaded bead-sliding velocity in a dose-dependent manner. With the antibody at 1.5-3 mg/ml, the beads eventually stopped moving to remain attached to actin cables. These beads could be readily detached from actin cables with very small centrifugal forces, indicating very weak actin-myosin linkages. The antibody had no effect on rigor actin-myosin linkages formed before the antibody application. These results are consistent with the view that myosin S-2 plays an essential role in muscle contraction. PMID- 9852348 TI - Tyrosine kinase-independent extracellular action of genistein on the CFTR Cl- channel in guinea pig ventricular myocytes and CFTR-transfected mouse fibroblasts. AB - The effects of genistein, a protein tyrosine kinase inhibitor, on the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel were studied in guinea pig ventricular myocytes and in NIH3T3 mouse fibroblasts stably transfected with CFTR cDNA by the whole-cell patch-clamp technique. Genistein did not activate whole-cell Cl- currents when applied to the intracellular (pipette) solution. In contrast, when applied to the extracellular solution, genistein alone promptly activated the Cl- current in a fully reversible manner. Also, extracellular genistein reversibly potentiated the forskolin-activated Cl- current. However, both basal and forskolin-activated Cl- currents were not affected by other protein tyrosine kinase inhibitors, including herbimycin A, lavendustin A, tyrphostin 21, tyrphostin 47, and tyrphostin 51. A nonspecific inhibitor of protein phosphatases, orthovanadate, had no effect on the genistein induced activation of CFTR. Pretreatment with a protein kinase inhibitor, either H-89 or H-7, or with an adenylate cyclase inhibitor, SQ 22536, also had no effect on the genistein-induced response. Thus, it is concluded that genistein alone activates CFTR by a protein tyrosine kinase-independent and protein phosphatase independent mechanism from the extracellular side, but not from the intracellular side. PMID- 9852350 TI - Distribution and cytological properties of macrophages in human Fallopian tubes. AB - The macrophages in human Fallopian tubes of women in the reproductive and postmenopausal periods were examined with a focus on their morphological properties by immunohistochemical staining and transmission electron microscopy. The fine structure of the smooth muscle cells in the Fallopian tubes was also investigated during the reproductive period. For immunohistochemical staining, we used two monoclonal antibodies that were specific for human macrophages, namely PM-1K and PM-2K. PM-1K recognizes human monocytes/ macrophages corresponding to CD68, and PM-2K recognizes tissue macrophages. PM-1K-positive cells were always present and their numbers increased significantly during the menstrual and early to mid-secretory phases. In contrast, relative numbers of PM-2K-positive cells were small throughout the menstrual cycle. In the postmenopausal period, few PM 1K-positive cells were detected, but PM-2K-positive cells remained. The macrophages during the secretory phase in the endosalpingeal stroma had well developed intracytoplasmic organelles, but relatively few cytoplasmic vacuoles and granules. In the same phase, many cells of the monocyte/macrophage lineage appeared in the vascular lumen of the endosalpingeal stroma. The macrophages during the menstrual phase had well-developed intracellular organelles, with cytoplasmic vacuoles and granules of various sizes and configurations. During the late secretory phase, just prior to menstruation, the smooth muscle cells contained few cytoplasmic filaments but electron-lucent or electron-dense lysosome-like bodies were seen. These findings suggested the presence of macrophages in human Fallopian tubes. It is possible that such macrophages might be involved in the physiological functions of the tubes during the reproductive period and moreover that they might participate in the reconstruction of the muscle layer of the tubes. PMID- 9852349 TI - Epithelial rests colocalize with cementoblasts forming acellular cementum but not with cementoblasts forming cellular cementum. AB - Epithelial rests of Malassez and cementoblasts were examined in the rat molars during the early stages of root formation using an antilaminin antibody and/or peanut agglutinin (PNA), and an antiosteocalcin (OC) antibody, respectively. The roots of the first molars were used for study. The antilaminin antibody stained the basement membrane surrounding the epithelial root sheath and epithelial rests. The basement membrane of the epithelial root sheath was continuous, but that of the epithelial rests was discontinuous. The cells of epithelial rests and epithelial root sheath were positive for PNA. The structural characteristics of the epithelial rests were seen in the sections stained doubly with PNA and the antilaminin antibody. The cells of epithelial rests were fibroblast-like and formed a fine mesh in 2-week-old rats. In 3-week-old rats, the epithelial rests were also present at the coronal half of root surface, showing typical cell cords, but were not present at the apical part of the root surface where the cellular cementum covered the root dentin. At the root apex of 3-week-old rats, the cells of epithelial rests forming fine meshes were seen near the epithelial root sheath. The anti-OC antibody stained cementoblasts lining acellular and cellular cementum. The sections doubly stained with the anti-OC and the antilaminin antibodies or PNA further revealed the close relation between epithelial rests and cementoblasts. The OC-positive cells lining acellular cementum or dentin were localized very close to the epithelial rests. In contrast, the OC-positive cells lining cellular cementum did not show close association with the epithelial cells, except the cells located most apically where the basement membrane of the epithelial root sheath is disrupted and the initial cellular cementum begins to be formed. The present results suggest that the epithelial rests and/or the discontinuous basement membrane of them may have a role for the acellular cementum formation at least in the early stage of root formation. PMID- 9852351 TI - Electron-microscopic detection of cytoskeletal actin in the epithelial cells of bovine abomasum. AB - The fine structural distribution of cytoskeletal actin was investigated in the oxyntic gland region and in the pyloric gland region of bovine abomasal mucosa using immunoelectron methods. In all exocrine cell types of the gastric epithelium, actin filaments exhibit a polarized pattern of distribution. In the members of the mucus- and protein-synthesizing cell line, the actin filaments form a prominent barrier to the exocytosis of secretory granules underneath the apical membrane. Therefore, actin is supposed to be involved in the control of exocytosis. The basolateral subplasmalemmal cortex of the exocrine cells, however, shows weaker labeling for actin filaments. In this position actin might be responsible for the movement of epithelial cells along the length of the gastric gland during physiological cell renewal and for the maintenance of the integrity of the epithelial unit and of cell polarization. In parietal cells of bovine abomasum, actin filaments are, in addition, associated with the intracellular canaliculus, but not with the tubulovesicular compartment. We assume that actin filaments reorganize the canalicular membrane after membrane translocations in the course of acid secretion. For the first time, actin filaments were documented in the rare brush cell type of bovine abomasum, where they do not form a terminal web beneath the apical membrane and therefore point out the nonexocrine function of the brush cell type. In all polarized cells of the bovine abomasum, actin filaments underlay the two apical members of the junctional complex, the tight junction and the adherens junction, where they might be involved in the regulation of cell adhesion, cell motility and cell shape determination. PMID- 9852352 TI - Adrenal microvascularization in the common tree shrew (Tupaia glis) as revealed by scanning electron microscopy of vascular corrosion casts. AB - The blood supply of the adrenal gland in the common tree shrew (Tupaia glis) was studied by use of transmission electron microscopy and vascular corrosion cast/scanning electron microscopy techniques. It was found that the gland receives its blood supply from branches of the inferior phrenic, aorta and renal arteries. Upon reaching the gland, these arteries divide into cortical and medullary arteries. The cortical arteries give rise to the subcapsular capillary plexuses which partially enclose the clusters of cells in the zona glomerulosa (ZG) and appear as lobular-like microvascular networks before running among the cellular cords in the zona fasciculata (ZF) and zona reticularis (ZR). It was noted that the capillaries in ZG and ZR are with more anastomoses than those in the ZF. Capillaries from the ZR become the sinusoidal capillaries in the adrenal medulla before proceeding to the peripheral radicles of the central vein. The medullary arteries penetrate the adrenal cortex and occasionally give off small branches to supply the inner cortex, especially the ZR. Their main branches break up into small or conventional capillaries in the adrenal medulla. These capillaries drain the blood into the peripheral radicles of the central vein and medullary collecting veins which proceed further into a very large central vein. The present findings illustrate that the adrenal medulla receives two blood supplies that yield somewhat different influences upon the adrenal medulla. The portal blood vessel could not be illustrated in the tree shrew adrenal gland. PMID- 9852353 TI - High-field magnetic resonance imaging of the developing human brain from the 10th to the 16th week of gestational Age. AB - In the present work, high-field magnetic resonance imaging (HF-MRI) was applied to study the developing human brain paying particular attention to the structures of interest in pathology of malformation. The aim of the work was to evaluate the possible application of HF-MRI to the analysis of brain development in the absence of some limits of conventional histological technique. Seven formalin fixed human fetuses of 50, 65, 70, 85, 110, 116 and 125 mm crown/ rump length (corresponding to a gestational age ranging from 10 to 16 weeks) were examined in an imager-spectrometer equipped with a 4. 7-tesla horizontal magnet with a 33-cm bore. In the brain of all the fetuses the telencephalic, mesencephalic and rhombencephalic vesicles were recognizable and an easy quantitative evaluation of the brain curvatures in the absence of distortion due to dissection was possible. Comparing fetuses at different gestational ages, the spatial modification of the different vesicles was evident. In fetuses at 16 weeks of gestational age, stratified compartments of the telencephalic wall were evident. The germinal zone and the cortical plate were visible: the germinal layer was identifiable as a hypointensity in the periventricular area. The subplate zone and the intermediate zone emitted a strong intensity signal. Our study demonstrates that HF-MRI can contribute to the study of the complex developmental events in the human brain from the 10th to 16th week of gestational age in a submillimetric scale of resolution. This technique can provide information about the morphology of the encephalic vesicles and their relations with the bone cavity that cannot be obtained with conventional methods and may be a useful adjunct to histological techniques. PMID- 9852354 TI - Blebs in the mouse cerebellar granular layer as a sign of structural inhomogeneity. 2. Posterior lobe vermis. AB - When the adult mouse cerebellum is perfusion-fixed with 70% ethanol, paraffin embedded, sectioned, and the sections are rehydrated, the granular layer wrinkles into an elaborate array of blebs. In the posterior lobe vermis these blebs are seen in both transverse and sagittal sections, are symmetrical about the midline, reproducible between neighboring sections and between individuals, and bear a constant relationship to the Purkinje cell bands as revealed by zebrin II immunocytochemistry. The data suggest that blebs represent individual cytoarchitectonic units and thus that the mouse cerebellum is subdivided into several thousand modules, each comprising >10(2) Purkinje cells and their associated interneurons and glial cells. PMID- 9852355 TI - Articular facets of the coracoclavicular joint in Koreans. AB - This study was performed to investigate the occurrence of the coracoclavicular joint in Koreans. Materials used in this study were paired clavicles and scapulae obtained from 102 adult Korean cadavers (61 males, 41 females) ranging from 18 to 97 years in age. The occurrence of the joint was identified by a definite articular facet on the conoid tubercle of the clavicle and also by one on the superomedial surface of the coracoid process of the scapula. To compare the morphometric differences between specimens with and without coracoclavicular joint, the clavicle length, scapular border length, glenoid length, coracoid height, coracoacromial (CA) arch height, coracoid slope, and acromion slope were measured on all bones examined. The coracoclavicular joint was found in 10 (9.8%) of 102 individuals examined. Among them, nine (8.8%) showed bilateral coracoclavicular joints, and a unilateral right joint was noted in one case (1.0%). No significant difference in the incidence of the joint was noted between the right and left side. The incidence of the joint in males, a frequency of 9.8%, was the same as that in females. The joint was not found in individuals under 40 years old. However, the joint was present in 9.5% of those aged 40-59 years, and in 11.4% of those aged 60 years or over. There was no significant difference between various measurements taken from specimens with and without a coracoclavicular joint. These findings suggest that the occurrence of the coracoclavicular joint is related to aging, but not to the size of the scapulae or the slopes and heights of some CA arch elements. PMID- 9852356 TI - Nonimmune hydrops fetalis: fetal and neonatal outcome during 1983-1992. AB - Prognostic factors for survival of 62 fetuses and neonates with nonimmune hydrops fetalis (NIHF) were studied retrospectively. Twenty-eight infants survived >/=28 days which is 45% for all fetuses and newborns diagnosed with NIHF and 61% for liveborns with unresolved NIHF. Univariate analysis identified that mortality was associated with the presence of >/=2 serous cavity effusions and a need for chest compressions at birth. Multivariate logistic regression analysis confirmed that the presence of >/=2 serous cavity effusions was significantly associated with mortality from NIHF <28 days after birth [OR = 48.2 (CI 3.6, 662.9) (p < 0.004)]. We conclude that, compared to published cases from the 1970s and early 1980s, survival of liveborns with NIHF seems improved. The decrease in stillbirths is more notable. The severity of hydrops at birth is the key determinant for survival. PMID- 9852357 TI - Beta-glucuronidase and hyperbilirubinemia in breast-Fed babies. AB - In the present study we analyzed breast milk samples from mothers of breast-fed, healthy, term babies with unexplained prolonged jaundice for beta-glucuronidase activity. The differences of enzyme activity were not statistically significant in comparison to a control group of nonjaundiced newborns. We conclude that beta glucuronidase is not the only cause of prolonged jaundice in neonates. PMID- 9852358 TI - CO2 reactivity of the cerebral hemoglobin concentration in healthy term newborns measured by near infrared spectrophotometry. AB - CO2 reactivity of cerebral hemoglobin concentration was studied in 16 healthy term neonates on days 1 and 4 after birth using the near infrared spectrophotometry (NIRS) technique. The aim was to establish data on the physiological range of CO2 reactivity in healthy newborns and to investigate the influence of postnatal age on it. The CO2 reactivity measured by NIRS is expressed as the change of the total cerebral hemoglobin concentration (tHbR) per change of CO2 tension in micromol/l/kPa. We evaluated CO2 reactivity during increases and decreases of transcutaneous CO2 partial pressure and found in our methodological setting the data of the increases more reliable. In all infants but 1 we found a tHbR on day 1 with a mean value of 8.19 micromol/l/kPa (-1.39 to 18.87), in all infants on day 4 with a mean value of 9.54 micromol/l/kPa (2.76 25. 88). There is a trend to higher values between day 1 and day 4 (difference = 2.25 micromol/l/kPa; p = 0.08). The noninvasive NIRS technique enabled us to test the cerebrovascular CO2 reactivity of the tHbR for the first time in healthy term newborns. Data on its physiologic range and variability are presented and compared to findings from ventilated infants and other age groups. As the CO2 reactivity might be an indicator for infants at risk of cerebral damage, it is necessary to have data on the physiological range of this parameter. PMID- 9852359 TI - Elastin of the umbilical cord arteries and its alterations in EPH gestosis (preeclampsia). AB - Edema, proteinuria, hypertension (EPH) gestosis is accompanied by an increase of collagen content and premature replacement of hyaluronic acid by sulfated glycosaminoglycans both in the umbilical cord arteries and in Wharton's jelly. The effect of EPH gestosis on elastin content and metabolism in the umbilical cord arterial wall was the aim of this work. Studies were performed on normal umbilical cord arteries and those taken from newborns of mothers with EPH gestosis. Elastin was isolated from the arterial wall and quantified by a dye binding method. Biosynthesis and degradation of this protein was evaluated by a pulse-chase experiment with the use of 14C-proline. It was found that EPH gestosis is associated with a significant reduction of elastin content in the umbilical cord arteries as a result of decrease in elastin biosynthesis rate and accelerated degradation of this protein. The replacement of elastin by collagen, and hyaluronate by sulfated glycosaminoglycans, may decrease the hydration of arterial wall and reduce its elasticity. Such rearrangement of extracellular matrix of the umbilical cord arteries may affect mechanical properties of these vessels and disturb fetal blood circulation. PMID- 9852360 TI - Measurement of cerebral oxyhaemoglobin saturation and jugular blood flow in term healthy newborn infants by near-infrared spectroscopy and jugular venous occlusion. AB - Data of cerebral haemodynamics and oxygenation are important for optimal treatment and prognosis in neonatal intensive care. Mostly premature and asphyxiated infants have been examined, but near-infrared spectroscopy (NIRS) allows estimations in healthy term newborns. In this study, cerebral venous saturation (CVsO2) and jugular blood flow (JBF) was estimated in 11 healthy term newborns. Mean CVsO2 was 64.12 +/- 4.6%, which conform with expectations. Mean JBF was only 6.1 ml/100 g/min, which is unacceptably low compared to earlier published data. We discuss physiological and methodological aspects and conclude that the combination of NIRS and venous occlusion appears to be a reliable method for estimation of CVsO2 in normally healthy newborns, whereas the reason for the failure of the method for estimation of JBF is unclear. PMID- 9852361 TI - Factors influencing pulmonary surfactant protein A levels in cord blood, maternal blood and amniotic fluid. AB - The purpose of this study was to evaluate factors that influence pulmonary surfactant protein A (SP-A) levels in cord blood, maternal blood and amniotic fluid, as well as to establish the normal range of serum SP-A. Labor significantly influenced cord blood SP-A levels. The SP-A levels in maternal blood after delivery were correlated with, but higher than those before delivery. There was a correlation between SP-A levels in amniotic fluid and cord blood. Neonatal serum SP-A was not correlated with maternal SP-A levels. The normal range of SP-A in cord blood was 2.7-21.7 ng/ml following cesarean section without labor, 4.8-50.2 ng/ml after labor at gestational weeks 36-38, and 12.2-44.6 ng/ml at gestational weeks 39-41. SP-A levels in maternal blood before and after delivery and amniotic fluid were 6. 0-74.0, 9.6-73.6 and 403.4-24,540 ng/ml, respectively. PMID- 9852362 TI - Effect of age and weaning on gastric mucosal injury in developing rats. AB - We evaluated ethanol- and HCl-induced mucosal damages in developing rats. The degree of damage induced by ethanol and HCl was greatest in 1-week-old rats and decreased significantly with age until 4 weeks; thereafter it increased again. To evaluate the effect of weaning on the maturational changes in mucosal defense, we compared ethanol-induced mucosal damage among three groups of newborn rats: (1) receiving milk only; (2) receiving only rat chow from 14 days of age, and (3) having free access to milk and chow. There were no significant differences at 18 and 21 days of age. The mucus thickness increased with age until 8 weeks and was not affected by weaning. In conclusion, developmental changes occur in gastric mucosal protection in rats. Weaning does not have a significant effect on these changes. PMID- 9852363 TI - High-frequency oscillatory ventilation, partial liquid ventilation, or conventional mechanical ventilation in newborn piglets with saline lavage-induced acute lung injury. A comparison of gas-exchange efficacy and lung histomorphology. AB - It has been reported that, in diseased lungs, either partial liquid ventilation (PLV) or high-frequency oscillatory ventilation (HFOV) can improve oxygenation better and with less lung injury than conventional mechanical ventilation (CMV). This study was intended as a preclinical comparison between the effects of HFOV, PLV and CMV on gas exchange, lung mechanics and histology. Fifteen anesthetized newborn piglets, with respiratory insufficiency due to repeated saline lung lavage, were allocated to either a PLV, HFOV or CMV (n = 5 each) strategy, and treated for 4 h. Within 30 min of commencing therapy, PLV, HFOV, and CMV improved arterial PO2 (Pa,O2), alveoloarterial oxygen gradient (P(A-a),O2), oxygenation index (OI), venous admixture (va), and arterial PCO2 (Pa,CO2). After 4 h, oxygenation parameters (Pa,O2, P(A-a),O2, OI and venous admixture) were significantly better in the HFOV group than in the PLV group; the CMV group showed a higher Pa,O2 and lower OI than the PLV group. Gas exchange at the end of the experiment was not different from baseline in the HFOV and CMV groups. Lung histology and morphometry were performed after perfusion-fixation at endotracheal deflation pressure corresponding to mean airway pressure at the end of the experiment. Lung injury score and mean linear intercept were not different between the three treatment groups. We conclude that in this model, gas exchange improved significantly in all three ventilation strategies. Indices of oxygenation improved less during PLV. The saline lavage-induced acute lung injury model used as in this study, is less stable than previously thought. The final lung injury is not influenced by the ventilation strategy. We speculate that the impaired gas exchange during PLV is an expression of diffusion limitation and ventilation-perfusion mismatch in a recovering lung. PMID- 9852364 TI - Effect of exchange transfusion on brain perfusion and electrocortical brain activity in newborn lambs. AB - Changes in brain perfusion (using ultrasonic-determined changes in carotid artery blood flow: Qcar) and electrocortical brain activity (ECBA, obtained by a Lectromed cerebral function monitor) were studied during exchange transfusion using the push-pull method in the newborn lamb. Changes in mean arterial blood pressure (MABP), Qcar, heart rate (HR), and ECBA were observed during all exchange transfusions: MABP, Qcar, and ECBA decreased during the withdrawal period and increased during the infusion period, whereas HR showed the opposite phenomenon. Changes in ECBA appeared to be primarily associated with changes in brain perfusion (p < 0.05). These changes in brain perfusion were, however, caused by changes in MABP (p < 0.0001). We concluded that exchange transfusions caused moderate changes in brain perfusion, however changes in ECBA appeared to be very small, and have probably no clinical importance. PMID- 9852365 TI - Diagnostic value of troponin T in neonates of mild pre-eclamptic mothers. AB - The measurement of myocardial damage by newer, highly specific markers of myocardial damage is now possible, including cardiac structural proteins such as troponin T (TnT). In neonates of pre-eclamptic mothers, it identifies minor myocardial damage missed by other biochemical markers. The present study was designed to determine the diagnostic value of TnT concentrations in neonates of pre-eclamptic mothers. Fifteen neonates of pre-eclamptic mothers were studied (9 boys and 6 girls), and 17 healthy full-term neonates (9 boys and 8 girls) were selected as a control group. The serum TnT concentration in neonates of pre eclamptic mothers (0.70 ng/ml) was significantly higher than that in the control group (0.10 ng/ml). In an echocardiographic study, the mean mitral peak velocity at an atrial contraction (A) value of 39 cm/s in neonates of pre-eclamptic mothers was significantly lower than that in the control group (53 cm/s), and the mean mitral peak velocity of early diastole to peak velocity of the atrial contraction (E/A) value (1.75) in neonates of pre-eclamptic mothers was significantly higher than that in the control subjects (1.23). In conclusion, our study demonstrated high levels of cardiac TnT, lower mitral A values and high mitral E/A values in neonates of pre-eclamptic mothers, presumably associated with mild myocardial damage in the neonates of pre-eclamptic mothers. PMID- 9852366 TI - Reliability of the Romanian version of the alcohol module of the WHO Alcohol Use Disorder and Associated Disabilities: Interview Schedule --Alcohol/Drug-Revised. AB - Alcohol Use Disorder and Associated Disabilities Interview Schedule - Alcohol/Drug-Revised (AUDADIS-A/D-R) is a fully structured, standardized and precoded instrument designed to evaluate alcohol and drug use disorders according to DSM-III-R, DSM-IV, and ICD-10 criteria. The AUDADIS-A/D-R has shown good to excellent reliability in both large clinical and general population samples, but prior to the conduct of the present study no data on the reliability of the Romanian version of the AUDADIS-A/D-R existed. The purpose of the present study was to examine the test-retest reliability of the alcohol module of the AUDADIS A/D-R in a general population and clinical sample in Romania. The overall reliability of ICD-10 and DSM-IV abuse, harmful and dependence diagnoses, was found to be good to excellent, but was somewhat lower for abuse and harmful use diagnoses. The results are discussed in terms of the cultural applicability of the symptom items and within the context of the analysis of discrepant responses between the test and retest interviews. PMID- 9852367 TI - Alcohol and disinhibition. AB - This review investigates research evaluating the disinhibition hypothesis. This hypothesis postulates that in a sober state behavior is inhibited. When people are influenced by alcohol the inhibitions are supposed to be weakened and the motivating drives are postulated to become disinhibited and potent to influence behavior. This report reviews the effect of alcohol on nerve functions, on human sexuality, aggression, eating behavior, psychological conflicts, fluency in talk, social anxiety, violent crimes and the interaction of alcohol and social norms. It has been shown that individual subjective experiences sometimes indicate disinhibition (reduction of the forces holding back impulses) and objective behavior in some respects was different when the subject was intoxicated, but the mechanism that mediates behavior is not clear. It seems to be difficult to measure independently the forces restraining (inhibiting) the driving forces (uncontrolled impulses) at the same time as measuring these driving forces. The review concludes that there is no unambiguous support of the disinhibition hypothesis. An alternative hypothesis that seems to explain many behaviors in an inebriated individual is the 'time out' hypothesis which states that drunken behavior is influenced more by norms about what it should be than by the pharmacological effect of alcohol. PMID- 9852368 TI - West German drinking patterns in 1984 and 1990. AB - Basic drinking patterns are examined with regard to sociodemographic correlates for the years 1984/1986 and 1990/1992 by a secondary analysis of data from representative samples of the former West German general population. Data come from the National Health Survey of the German Cardiovascular Prevention Study in which the former West German non-institutionalized general population aged 25-69 years was sampled through multistage probability methods. In 1984, 91% of West Germans were drinkers, 16% were daily drinkers, and 8% consumed a mean of 60 g or more of ethanol per day. These figures decreased slightly but significantly by 1990. Despite a downward trend in consumption between 1984 and 1990, Germany remains a 'wet' culture. PMID- 9852369 TI - Drug policies and harms: a conceptual framework. AB - Existing indicators of drug-related policies and harms - official statistics, treatment data, and so on - are difficult to interpret and to compare across jurisdictions. We propose a conceptual framework including key dimensions of policy and harm. The framework is designed to guide creation of a new dataset in which existing empirical data can be used to assign city-specific scores on policy and harm dimensions. By an interpretive process involving local experts and a coordinating body, we believe that existing data can be evaluated and synthesized to derive these scores. We also propose a cumulative case-study method of querying the dataset to test hypotheses regarding relationships between policies and harms. PMID- 9852370 TI - Drugs on prescription--the Swedish experiment of 1965-67 in retrospect. AB - The programme of prescribing amphetamines and opiates to drug users in Stockholm carried out from 1965 to 1967 has frequently been referred to in the European drug policy debate. In this article, originally presented at the Conference on Drug Use and Drug Policy in Amsterdam in September 1996, the programme is described with regard to design, setting and outcome. The relevance of the programme for the general policy debate is discussed and a comment is given on the programme's impact on the Swedish drug epidemic and Swedish drug policy. PMID- 9852371 TI - Characteristics of long-term cannabis users in Sydney, Australia. AB - Two hundred long-term cannabis users (58% male) were interviewed on their characteristics and experience of use. Respondents had been regularly using cannabis for an average of 11 years and more than half used daily (56%). The most common route of administration was in a waterpipe, and nearly all (93%) smoked the flowering heads ot the plant. One in 5 (21%) had a cannabis-related conviction. The benefits of use were perceived to be its relaxing, mood-enhancing effects, and its ability to alter consciousness. The most commonly cited negative aspects of use were cost, negative psychological effects and legal status. Polydrug use was common, with alcohol and tobacco almost universally used on a regular basis. More than half the drinkers in the sample were consuming alcohol at hazardous or harmful levels. PMID- 9852373 TI - German research network 'Analytical Epidemiology of Substance Abuse' (ANEPSA). PMID- 9852372 TI - Outpatient opiate detoxification treatment with buprenorphine. Preliminary investigation. AB - In an open study design, 50 opioid-dependent subjects (DSM-IV: 304. 0) were investigated in a gradual detoxification treatment with buprenorphine. The study was performed at the drug addiction outpatient clinic of the Department of General Psychiatry at the University of Vienna. Subjects had to contact the outpatient clinic on a daily basis and buprenorphine was administered according to their clinical status. Withdrawal symptoms were evaluated by applying the WANG scale. Urine samples were screened for drug toxicology to exclude additional consumption. In this investigation buprenorphine was applied sublingually in a free dosage scheme aimed at completing detoxification treatment within 10 days by reducing buprenorphine on a daily basis. A mean daily dosage of 2.3 mg buprenorphine was required by patients on day 1 of the treatment period. The highest mean daily buprenorphine dosage was given on day 2, followed by a daily reduction over the study period. The result of this open study design revealed that a gradual daily reduction of buprenorphine might be a successful alternative outpatient detoxification treatment in opioid-dependent subjects. Compliance was 70%, the reported and evaluated withdrawal symptoms during the study period were moderate. PMID- 9852375 TI - Age increases leucine enkephalin hydrolysis in human plasma. AB - BACKGROUND: The existence of age-associated alterations in immune functions and neuropeptides capable of modulating these functions prompted us to advance the hypothesis that the degradation of plasma neuropeptides, specifically opioid peptides, may be altered by aging. OBJECTIVE: To verify the possible existence of age-induced variations in neuropeptide hydrolysis in human plasma, using leucine enkephalin as the model substrate. METHODS: The hydrolysis of leucine enkephalin and the formation of its hydrolysis byproducts in the presence of plasma enzymes were studied by kinetic and chromatographic techniques in a group of elderly individuals and a control group. RESULTS: The results obtained indicate that in elderly individuals the activity of enkephalin-degrading plasma enzymes is greater than in controls. ANOVA analysis of these data indicates that the dependency of the variation of hydrolysis upon the 2 age groups is statistically significant. Increased substrate hydrolysis, and a modified hydrolysis pattern, appear to be associated with increased activity of the enzymes involved, and with different distribution of the individual enzymes within each class, as well as with severely reduced activity of the low molecular weight plasma inhibitors. CONCLUSION: The combination of the above-mentioned factors appears to define a characteristic hydrolysis pattern for elderly individuals which is different from that found in controls. PMID- 9852374 TI - Toward understanding age-related memory loss in late adulthood. AB - BACKGROUND: While laboratory tests indicate that older adults typically perform more poorly than do younger adults on many types of memory tasks, the question arises as to whether, or to what extent, it is valid to attribute these differences to ageing per se or to some variable or class of variables that intervene between age and remembering. OBJECTIVE: The purpose of this review is to present three current views that might explain the relationship between age and remembering. They can be construed as variants on resource theories and include: the processing speed hypothesis, the executive function hypothesis, and the common cause hypothesis. METHODS: The review samples results pertinent to these hypotheses that derive from behavioural research. Studies involving various imaging techniques were considered beyond the scope of the review. RESULTS: The balance of research strongly implicates reductions in the speed of information processing as a fundamental contributor to normal age-related memory loss. Nonetheless there are circumstances where other mechanisms, such as working memory, executive function, and sensory processes, are important. CONCLUSION: Despite the phenomenological and empirical reality of age-related memory loss and the breadth of attempts to explain it, much work remains to be done to understand why it occurs. Contemporary debates about the nature and means of identifying shared and unique effects promise to shape future directions for research on memory aging. PMID- 9852376 TI - How age changes the content of neuroendocrine peptides in the murine gastrointestinal tract. AB - BACKGROUND: Motility disorders in the gastrointestinal tract increase with ageing, and colorectal carcinoma is a tumor of the middle-aged and elderly. Gastrointestinal secretion, absorption, motility, cell proliferation, local immune defense and blood flow are all regulated by the neuroendocrine peptides. It is conceivable that gastrointestinal disorders at an advanced age may be accompanied by changes in this regulatory system. OBJECTIVE: To ascertain possible age-related changes in neuroendocrine peptides in a rodent animal model. METHODS: The concentrations of various neuroendocrine peptides were determined by radioimmunoassays in tissue extracts from the antrum, duodenum and colon of mice in four different age groups: 1, 3, 12 and 24 months. The neuroendocrine peptides investigated were: secretin, gastric inhibitory polypeptide (GIP), gastrin, motilin, peptide YY (PYY), somatostatin, vasoactive intestinal polypeptide (VIP), substance P, neuropeptide Y (NPY), galanin and neurotensin. RESULTS: Antrum: Concentrations of somatostatin, VIP and substance P decreased significantly in 1 month-old mice (but that of neurotensin increased) compared with 3-month-old mice. In 12-month-old and 24-month-old mice, concentrations of gastrin, somatostatin, VIP, substance P, NPY, galanin and neurotensin all decreased vis-a vis 3-month-old mice. Duodenum: Whereas the levels of secretin, GIP and neurotensin increased, those of gastrin, motilin, somatostatin, VIP, substance P, NPY and galanin decreased in 1-month-old mice vis-a-vis 3-month-old mice. In both 12-month-old and 24-month-old mice, the concentrations of secretin and GIP increased, compared with those of 3-month-old mice. The levels of gastrin, motilin, somatostatin, VIP, NPY and galanin decreased in both 12-month-old and 24 month-old mice vis-a-vis 3-month-old mice. Substance P and neurotensin concentrations decreased in 12-month-old mice, but not in 24-month-old mice. Colon: In 1-month-old mice the levels of PYY, somatostatin, VIP, substance P and galanin decreased vis-a-vis 3-month-old mice. In 12-month-old mice, the concentrations of PYY, somatostatin, VIP, NPY, galanin and neurotensin decreased compared with those in 3-month-old mice. In 24-month-old mice, the VIP level decreased, whereas the substance P level increased. CONCLUSION: The changes in neuroendocrine peptides observed in the gastrointestinal tract of this murine animal model could be of some relevance for the increased gastrointestinal dysfunction in the elderly human. They may also be involved in the development of colorectal cancer. PMID- 9852377 TI - Effect of in vitro aging on the modulation of protein and fibronectin biosynthesis by the elastin-laminin receptor in human skin fibroblasts. AB - The 67-kD elastin-laminin receptor (ELR) subunit which carries the recognition site for elastin peptides (EP) is a lectin. Its binding with galactosides can modulate the kinetics of its interaction with its ligand, EP. In this study the biosynthesis of proteins, collagen and fibronectin were evaluated in the presence of agonists and antagonists of the receptor on human skin fibroblasts. The biosynthesis of total proteins determined by 3H-proline incorporation and of fibronectin (by immunoprecipitation) were shown to increase with passage number. The presence of 1 microg/ml kappa-elastin (EP) in the culture medium increased both total proteins and fibronectin biosynthesis. Melibiose, an agonist of the receptor at 5 microg/ml (140 microM), decreased both proteins and fibronectin biosynthesis in the culture medium of human skin fibroblast at the 10th and 15th passage. These results show that the ELR can control the biosynthetic mechanisms of some of the macromolecular constituents of extracellular matrix such as fibronectin and moderate its age and passage-dependent upregulation. PMID- 9852378 TI - Antibody response to 1995-1996 influenza vaccine in institutionalized and non institutionalized elderly women. AB - BACKGROUND: Concern about poor responsiveness to influenza vaccination by institutionalized elderly people. OBJECTIVE: To determine whether institutionalized elderly volunteers develop a significant antibody response following influenza vaccine and to compare this response with that of non institutionalized subjects. METHODS: The haemagglutination-inhibiting antibody response after 1995-1996 influenza vaccination [A/Shangdong/9/93 (H3N2), A/Taiwan/1/86 (H1N1), B/Panama/45/90] was estimated in 80 elderly women living in a nursing home and compared with that of 51 non-institutionalized women. RESULTS: No differences were found in the prevaccination status, and, after vaccination, a significant humoral response was elicited both in institutionalized and non institutionalized elderly subjects against all three influenza strains tested. The immune response of institutionalized patients was satisfactory and significantly higher than that observed in non-institutionalized women. These results were confirmed both by a separate analysis of homogeneous subgroups stratified according to the presence in the two cohorts of potential causes of differential antibody response (prevaccination antibody titre, age, long-term drug treatment, risk factors for influenza infection, and physical disability) and by logistic regression analysis in order to adjust immune responses for the different variables. CONCLUSION: Influenza vaccination is effective in elderly people living in nursing homes. However, the postvaccination antibody response to influenza vaccine is influenced by different factors directly or indirectly related to residence. PMID- 9852379 TI - Assessment of body composition in elderly: accuracy of bioelectrical impedance analysis. AB - BACKGROUND: In the literature there are several body impedance analysis (BIA) prediction equations generally determined in younger populations and their accuracy in the elderly has not been adequately confirmed. OBJECTIVE: We verified the reliability of the BIA method in a body composition study in the elderly. METHODS: To assess the accuracy of bioelectrical impedance analysis we compared this method with dual photon absorptiometry (DPA), assumed as a gold standard; body composition was predicted by seven BIA prediction equations in 24 healthy elderly individuals. RESULTS: The best equation in fat-free mass (FFM) estimation is the RJL System formula (published by Segal in 1988); nevertheless, the wide range of the error in FFM estimating may limit its clinical application. The FFM hydration variability seems to be the principal variable which explains the error in FFM estimation by BIA prediction equations. CONCLUSION: These findings indicate that the use of BIA equations is not interchangeable, when FFM is calculated in an elderly population, and more validation studies are necessary in this age group to evaluate the clinical application of this method. PMID- 9852380 TI - A neuropathological study of dementia in nursing homes over a 17-year period, in Shimane Prefecture, Japan. AB - BACKGROUND: In Japan, vascular dementia (VD) has been responsible for the majority of all dementia cases in both epidemiological and neuropathological studies. Recently, however, several epidemiological investigations have shown that the rate of VD has decreased and senile dementia of the Alzheimer type (SDAT) is now the major cause of dementia, though few neuropathological investigations show that the majority of dementia is SDAT. OBJECTIVE: The purpose of the study was to clarify the rates of dementia types in Japanese nursing home residents and to compare the ratio of SDAT to VD in the first and second halves of the study. METHODS: One hundred and twenty-two demented nursing home residents (48 men and 74 women) were evaluated neuropathologically over a period of 17 years in Shimane prefecture, Japan. All subjects died between 1976 and 1992. The average age at death was 81.4 +/- 9.4 years for men, 85.0 +/- 7.2 for women and 83.6 +/- 8.3 for total samples. RESULTS: In classifying dementia type, SDAT accounted for 34% (41 cases); VD 35% (42); mixed dementia 11% (14); and 'other' dementia 20% (25) of all samples. Comparison of the first 9-year period with the second 8-year period revealed the ratio of SDAT to VD tended to increase (from 0.83 to 1.15). This increasing tendency was found both in men (from 0.53 to 0.7) and in women (from 1.13 to 1.44). These tendencies, however, were not statistically significant. CONCLUSION: There was no change statistically in the ratio of SDAT to VD between the first half of the study and the second half. However, there was an increasing tendency of the ratio in our study. PMID- 9852381 TI - Oral flora of elderly patients following acute medical admission. AB - The human oral microflora is diverse and is usually predominately composed of Gram-positive bacteria. It is uncommon to find Gram-negative bacilli (GNB) in healthy mouths. The incidence of infection with GNB rises in institutionalised, frail elderly subjects. There is also evidence of an association between intra oral GNB presence and denture wearing. There have been few studies which have investigated intra-oral GNB carriage in acutely ill elderly patients. The aim of this study was to evaluate the oral flora of a group of elderly patients during an acute medical admission and to investigate any associations between the oral microflora and existing medical or oral factors. A total of 28 patients (17 females and 11 males; age: 74-93 years) on a care for the elderly ward were studied. Epidemiological data, detailed medical histories and oral examinations were undertaken. In addition, oral swabs of the palate area were taken to determine their oral flora. Twelve (43%) of the patients had GNB in their oral cavities. These patients were suffering from a variety of medical conditions and were on various drug regimes. There was a correlation between oral GNB presence and denture use. There was no association between GNB presence and denture hygiene. As oropharyngeal GNB colonisation can be associated with infections such as aspiration pneumonia, it is important in patients at risk that intra-oral organisms are identified and managed. PMID- 9852382 TI - Effect of aging on cardiothoracic ratio in women: a longitudinal study. AB - BACKGROUND: The effect of aging on the components of the cardiothoracic ratio (CTR) in the chest roentgenogram in women remains obscure. There have been only a few reported studies that have investigated longitudinal changes of the CTR in women. OBJECTIVE: To determine whether there is an effect of aging on the CTR in women. METHODS: The study population consisted of 110 women who attended an annual health screening and underwent chest roentgenography both in 1981 and 1990 in a mountain village in Japan. We used the data obtained from an annual health screening in order to determine thoracic diameter (TD), cardiac diameter (CD), and lung height (LH) and then calculated the CTR. We analyzed CTR, TD, CD, and LH on the basis of the age of the participants. RESULTS: In the cross-sectional analysis of the groups stratified by age in 1981, mean CD and CTR increased, and TD decreased in the advanced-age group. The percentage of radiographic cardiomegaly (CTR >50%) strikingly increased in the advanced-age group. In the longitudinal analysis between 1981 and 1990, the mean CD increased, and the mean TD decreased significantly. The mean CTR increased by 2.0% (95% CI 1. 2-2.8%). The changes of the CTR in 9 years were positively correlated with those of CD and negatively correlated with those of TD and LH. The longitudinal data generally accorded with the cross-sectional data, except LH. CONCLUSIONS: The CTR in women increases with aging as a result of the increase in CD, except in the case of advanced old age, and the decrease in TD. LH could be another major factor that has an influence on CTR. Caution should be exercised to avoid overestimating CTR when LH changes. The classical upper limit of 50% for CTR in our adopted method should not be applied to elderly women because of the possibility that changes with normal aging are misclassified as abnormal findings. PMID- 9852383 TI - Upper body obesity and hyperinsulinemia are associated with anovulation. AB - The objective of this study was to determine the effect of body fat distribution and hyperinsulinemia on the occurrence of ovulation. Fifty-six women (20-35 years old) either with overweight or obesity (body mass index >/=25) were studied. They were classified in two groups according to waist/hip ratio (WHR); one with predominance of adiposity in the upper body segment (n = 29, WHR >0.85) and the other with predominant adiposity in the lower body segment (n = 27, WHR 4 (p < 0.002), para > 3 (p < 0.01) and previous caesarean section (p < 0.02). There is no increase in the incidence of placenta previa with increasing maternal age and previous abortion. CONCLUSION: The risk of placenta previa is increased with higher gravidity, higher parity, previous lower segment caesarean section, while increasing maternal age and previous abortion had no significant effect. PMID- 9852385 TI - Diurnal variation in excretion of N-acetyl-beta-glucosaminidase during pregnancy: implications for the prediction of pre-eclampsia. AB - Significant differences in urinary excretion of N-acetyl-beta-glycosaminidase (NAG) were observed between nulliparous and multiparous patients and between fasted and nonfasted individuals. No significant difference was observed between patients with normal and those with abnormal glucose tolerance. Multiple regression analysis confirmed that the significant independent determinants of NAG/creatinine ratio were age, parity, gestation and fasting state. Significant diurnal variation in urinary NAG/Cr ratio was observed, the highest levels being recorded in early morning fasting specimens, falling in each postprandial specimen and beginning to rise again by midnight. The urinary NAG/Cr ratio is influenced by fasting, parity, gestation and age. More consistent results for prediction of pre-eclampsia are therefore likely to be obtained using fasting (early morning) urine specimens and adjusting cut-off criteria for the other factors. PMID- 9852386 TI - Clinical experience of long-term transdermal treatment with nitric oxide donor for women with preeclampsia. AB - Isosorbide dinitrate (ISDN), a nitric oxide donor, was applied transdermally for 4-16 days to 4 preeclamptic women with oligohydramnios, intrauterine fetal growth retardation (IUGR), and elevated resistance of blood flow in the uterine arteries. Pulsed Doppler ultrasonography revealed immediate and drastic improvement of pulsatility index (PI) of uterine arteries following treatment with ISDN. The average PI in uterine arteries of the 4 patients was reduced to approximately 67% of that of the untreated state. In 2 patients the amniotic fluid gradually increased over a few days which suggested improvement of fetoplacental circulation during administration of ISDN. This study suggests that long-term transdermal ISDN is an effective therapy, at least in a portion of preeclamptic women, to avoid maternal hypertension, fetal distress, oligohydramnios, and IUGR, and consequentially to prolong the gestational period. PMID- 9852387 TI - Ultrasonographic blood flow measurement in the carotid arteries in postmenopausal women. AB - OBJECTIVES: The aim of the present study was to evaluate blood flow in postmenopausal women on hormone replacement therapy (HRT) compared to controls. Blood flow was ultrasonographically measured in the great arteries of the neck instead of in the vessels of the internal genital organs. METHODS: Fifty healthy women with climacteric complaints, at least 6 months postmenopausal, participated in the study. They were randomly divided into two groups. One group received 2 mg estradiol (E2) for 12 days, continued with 2 mg E2 and 1 mg norethisterone acetate for 10 days, followed by 1 mg E2 for 6 days, cyclically during 6 months. The other group received placebo tablets the first 3 months and the same HRT as the first group for the last 3 months. Blood flow was measured ultrasonographically by color flow pulsed Doppler in the common (CCA), internal (ICA) and external (ECA) carotid arteries, before the start of the study, after 3 and 6 months of therapy. RESULTS: CCA and ICA, both low resistance vessels, and ECA, a high resistance vessel, and their waveforms were identified. Pulsatility index did not decrease statistically significant (p > 0.05) in any of the great vessels during 6 months of HRT in this study. There were no differences in blood flow between the HRT-treated group compared to control group during 3 months of therapy, except for the right ECA (p = 0.04). CONCLUSION: The difference in blood flow and wave-forms of the major arteries of the neck were clearly shown, but HRT did not have any important impact on the blood flow in this study. No difference was shown concerning blood flow between the two groups of postmenopausal women, on active therapy or placebo. PMID- 9852388 TI - Endometrial response to unopposed estrogens remains unaltered in patients with chronic renal failure receiving hemodialysis. AB - Most patients with chronic renal failure who are on maintenance hemodialysis are anovulatory and have menstrual abnormalities. This study was designed to determine the prevalence of organic causes of abnormal uterine bleeding in this group of patients exposed to unopposed estrogens. Eighteen patients with chronic renal failure and abnormal uterine bleeding underwent vacuum curettage. The histopathologic findings were compared with a group of 154 premenopausal women who had abnormal uterine bleeding without detectable organic causes. Excluding patients with secretory and atrophic endometrium, only 2 of 8 patients (25%) with chronic renal failure had endometrial lesions while 44 of 131 patients (33.6%) had either endometrial polyp, simple or atypical endometrial hyperplasia or endometrial carcinoma (p > 0.05). The uremic environment caused by chronic renal failure does not alter the endometrial responsiveness to unopposed estrogens and may lead to the development of endometrial lesions. PMID- 9852389 TI - Memory functioning at menopause: impact of age in ovariectomized women. AB - Estrogens are known to act selectively on some components of memory, exerting beneficial effects on cognitive performances. However, there are few data on the long-term effect of the lack of estrogen in postmenopausal women. Therefore, we investigated attentive and verbal memory performances in physiological and surgical menopause, drawing attention to the impact of age at menopause, and we compared a well-known aging and estrogen-dependent index, the entity of bone mass loss to memory functioning. No significant differences were found in the mean scores of attentive and psychomotor performances between physiological and surgical menopause, whereas a lower number of recalled words (recency effect = PS2) was found in surgical menopause (p < 0.001) in comparison to physiological menopause. In addition, both the age at the time of ovariectomy (r = 0.47; p = 0. 014) and the years since surgery (r = -0.64; p = 0.000) correlated to short-term verbal memory performance (PS2) with better scores when surgery occurred later in women's lives. Surgical menopause is able to affect short-term verbal memory more than physiological menopause and seems to represent a critical negative event within the female brain, in particular when it occurs prematurely. PMID- 9852390 TI - Effects of add-back therapy on bone mineral density and pyridinium crosslinks in patients with endometriosis treated with gonadotropin-releasing hormone agonists. AB - Treatment of endometriosis with gonadotropin-releasing hormone agonists (GnRHa) is limited to 6 months because of possible adverse effects on bone metabolism. We designed a randomized, double-blind, placebo-controlled, prospective study of 27 patients with endometriosis who were given GnRHa with or without hormone add-back therapy (+ 20 microg of ethinyl estradiol with 0.15 mg desogestrel) designed to suppress the adverse effects of hypoestrogenism while preserving the efficacy of GnRHa. Both regimens showed significant improvements in endometriosis, dysmenorrhea, and pelvic pain; effects were significantly better in the GnRHa + placebo group. The GnRHa + placebo group had significantly higher serum calcium levels and a significantly higher loss of lumbar spine bone mineral density (BMD). Urinary levels of pyridinium crosslinks increased significantly in the GnRHa + placebo group, and declined to normal in the GnRHa + add-back group. The add-back therapy protects women taking GnRHas from severe loss of BMD and accelerated bone collagen resorption, but reduces the efficacy of the GnRHa. PMID- 9852391 TI - Urinary bacterial flora of women with urethral syndrome and interstitial cystitis. AB - The aerobic and anaerobic bacterial flora in the first voided and in the midstream urine of healthy females (n = 5) and female patients with either urethral syndrome (US) (n = 5) or interstitial cystitis (IC) (n = 14) were studied. Bacteria were grown on media enabling isolation of fastidious and aerobic as well as obligatory anaerobic species. In healthy females only gram positive rods were found whereas US patients also harbored streptococci. Patients with IC presented also with Enterobacteriae and anaerobic bacteria. Five IC patients with severe symptoms were treated with metronidazole; 2 out of 3 patients with anaerobic bacteria in the pretreatment specimens had no anaerobes after metronidazole therapy and in 1 patient streptococci disappeared after the therapy. One patient with severe symptoms and Bacteroides fragilis in the midstream urine became symptom-free after 2 weeks of metronidazole treatment. Although there is uncertainty whether the US and IC are not related to an infectious etiology, the bacterial flora in urethral and in midstream urine in these conditions differs considerably from that of healthy females. PMID- 9852393 TI - Bilateral oophorectomy in asymptomatic women over 50 years old selected by ovarian cancer screening. AB - The purpose of this study was to evaluate bilateral oophorectomy in women over 50 years old found to have an adnexal mass using transvaginal ultrasonography (TVS) as a mass screening. With TVS a total of 23,451 women without symptoms were examined for ovarian cancer at annual screening for uterine cervical cancer. Two hundred fifty-eight women over 50 years old persistently had abnormal TVS results and 95 women gave informed consent for surgical tumor removal. In the 95 women operated, 7 malignant ovarian cancers were found. Especially adnexal masses which were thought to be benign were treated by laparoscopic surgery. PMID- 9852392 TI - Preoperative determination of several serum tumor markers in patients with primary epithelial ovarian carcinoma. AB - This study was designed to evaluate the clinical significance of the use of preoperative serum tumor markers in primary epithelial ovarian cancer. Subjects comprised 111 patients with primary epithelial ovarian cancer. Lactate dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (HBDH), carcinoembryonic antigen (CEA), CA19-9, tissue polypeptide antigen (TPA), CA125 and sialyl TN (STN) serum levels were measured within 7 days before surgery. The tumor marker values were compared with the histopathologic diagnosis. The overall agreement between the test results and the actual outcome was calculated using Student's t test and analysis of variance (ANOVA). Survival curves were constructed according to the Kaplan-Meier method, and differences in survival were assessed with the log-rank test. The prognostic significance of tumor markers for survival was assessed in a multivariate analysis with the Cox proportional hazards model. Of the tumor markers examined in this study, CA125 showed the highest positive rate (77.6%), followed by 63.2% for STN and 55.9% for CA19-9. When the positive rate was compared according to histologic types, serous cystadenocarcinoma, mucinous cystadenocarcinoma, endometrioid adenocarcinoma and clear cell carcinoma showed the highest positive rates for CA125 (94.1%), CA19-9 (76.9%), CA125 (91.7%) and STN (75.0%), respectively. Regarding the distribution of tumor marker levels according to the FIGO stage, LDH, HBDH, TPA and CA125 were correlated with the clinical stage while CEA, CA19-9 and STN did not show any correlation. From analyses of tumor marker levels according to histologic types, all patients with a ratio of CA125 to CEA of >1, 000 had serous cystadenocarcinoma and a ratio of CA125 to CA19-9 of >50 showed serous cystadenocarcinoma or endometrioid adenocarcinoma. On the other hand, all patients with a ratio of LDH or HBDH to CA19-9 of <1.0 had mucinous cystadenocarcinoma or clear cell carcinoma. From univariate analysis, the survival time of patients with elevated CA125, TPA or STN was significantly shorter than that of patients with normal CA125, TPA or STN levels. When the Cox's proportional hazard model was used, we identified age, clinical stage, clear cell carcinoma and serum STN as independent prognostic factors. Serum CA125, TPA or STN may give significant prognostic information in epithelial ovarian carcinoma. It is noteworthy that STN has been identified as an independent prognostic factor and has a high rate of positivity in clear cell carcinoma. PMID- 9852394 TI - Sacrospinous fixation for the prolapsed vaginal vault. AB - In 54 patients, transvaginal sacrospinous ligament fixation procedures were reviewed retrospectively. The mean operation time was 15 (12-45) min. The mean blood loss was 126 (110-175) cm3. The only intraoperative complication was a rectal laceration that was repaired primarily. The mean duration of follow-up was 28 (4-54) months. There were only 2 recurrent vaginal vault prolapses. There were 3 cases of cystocele (5.5%), 1 case of rectocele (1.8%), 5 cases of enterocele (9.2%), 3 cases of stress incontinence (5.5%), and 5 cases of dysparonia (9.2%). Sacrospinous ligament fixation can be used as an alternative treatment to vaginal hysterectomy in aged women with medical problems and young women suffering from genital descent with infertility. The procedure has the advantage of avoiding laparatomy, facilitating other vaginal repairs needed during the same operation, preserving vaginal function and shortening the time necessary for anesthesia and surgery. PMID- 9852395 TI - Malignant mixed mesodermal tumors of the ovary: preoperative diagnosis. AB - Early diagnosis of malignant mixed mesodermal tumors of the ovary is very difficult because of the rarity and the insidious onset. The purpose of this report is to review the magnetic resonance imaging features of an ovarian malignant mixed mesodermal tumor, which occurred in a 52-year-old woman, aiding in the differential diagnosis. PMID- 9852396 TI - Molecular mechanism of a cross-talk between estrogen and growth-factor signaling pathways. AB - The actions of estrogen (E2) are considered to be mediated through its nuclear E2 receptor (ER). In cancer development, growth factors are shown to act synergistically with E2. Recently, we found that the mitogen-activated protein kinase, activated by growth factors, phosphorylates human ERalpha and this phosphorylation potentiates the transactivation function of human ERalpha demonstrating a novel cross-talk between E2 and growth factor-signaling pathways. In this review, the molecular mechanism of this cross-talk is discussed. PMID- 9852397 TI - Genetic and epigenetic alterations of the estrogen receptor gene and hormone independence in human breast cancer. AB - The existence of hormone-independent tumors is a substantial problem for the present endocrine treatment of breast cancers. Estrogen receptor (ER) gene mutation can change the biochemical activity of the protein and can affect hormone responsiveness. However, quite a few mutations of significance have been described in breast cancer. Recently, numerous variant ERs have been detected at the mRNA level with alternative splicing, yielding deletion of exon 3, 5, or 7. The truncated ER protein induced from variant mRNA could mainly be exhibited as a repressor through dominant negative effects on normal ER protein. The mechanism of the loss of hormone dependency is, however, still very complex. Further work to assess the correlation between clinical behavior and ER variants is required to determine whether these variants play a role in hormone-resistant disease. Additionally, the DNA methylation of the ER gene itself may control ER expression. These epigenetic changes can play an important role in the loss of hormone dependence in breast cancer. PMID- 9852398 TI - Endocrine therapy in pancreatic carcinoma. AB - There are indications of the possible effects of sex hormones on pancreatic carcinoma. Estrogen receptor (ER) has been demonstrated in pancreatic tumors in experimental animals and in humans and it has been suggested that endocrine manipulation may be effective in the treatment of pancreatic carcinoma. However, it is still controversial whether this lethal cancer can potentially benefit from endocrine therapy. One explanation for the conflicting data on the benefit of hormonal manipulation in the treatment of pancreatic carcinoma may stem from the fact that there is no adequate marker to assess estrogen dependency of the pancreatic tumors. In this article, we review our work on tissue plasminogen activator as a prognostic guide to evaluate the efficacy of hormonal therapy in human pancreatic carcinoma, and also suggest that a selected subgroup of patients with this lethal cancer may have a potential clinical benefit from endocrine therapy, especially medroxyprogesterone acetate treatment. PMID- 9852399 TI - A pure antiestrogen, ICI 182,780, stimulates the growth of tamoxifen-resistant KPL-1 human breast cancer cells in vivo but not in vitro. AB - The critical mechanisms responsible for antiestrogen resistance have not yet been elucidated. We previously established a breast cancer cell line, KPL-1, derived from a patient with recurrent disease which appeared under tamoxifenadministration. In a previous study, we suggested that this cell line is estrogen receptor (ER)-positive but tamoxifen-resistant. In the present study, the effects of a pure antiestrogen, ICI 182,780, on this cell line were investigated. Although tamoxifen inhibited neither cell growth nor estradiol stimulated transcriptional activity in vitro, ICI 182,780, significantly inhibited both of them. Tamoxifen and ICI 182,780 were then administered to female nude mice bearing KPL-1 tumors. Tamoxifen had no effect on tumor growth, but ICI 182,780 unexpectedly stimulated it (p = 0.022). Estradiol tended to inhibit tumor growth (p = 0.198). Immunohistochemical analysis revealed that ICI 182,780 significantly increased the Ki6-labeling index (p<0.001) but estradiol decreased it (p = 0.035). To explore the possible mechanisms of these phenotypes, the mRNA levels of ER-alpha,ER-beta, transforming growth factor-beta1, fibroblast growth factor (FGF)-1 and FGF-4 in KPL-1 cells were compared with those in other ER-positive human breast cancer cell lines by reverse-transcription polymerase chain reaction. FGF-1 was overexpressed only in KPL-1 cells. This cell line is the first breast cancer cell line to be growth-stimulated by ICI 182,780 in vivo. Paracrine interaction between tumor cells and stromal cells mediated by growth factors, such as FGF-1, might be a key factor to explain the unique hormone responsiveness of KPL-1 cells. PMID- 9852400 TI - Role of estrogen and estrogen-related growth factor in the mechanism of hormone dependency of endometrial carcinoma cells. AB - The role of estrogen and estrogen-related growth factors in the mechanism of hormone dependency of endometrial adenocarcinoma cells was investigated. The proliferation of hormone-responsive human endometrial adenocarcinoma cells (Ishikawa cells), which possess both estrogen and progesterone receptors, was optimally stimulated by 10 nM estradiol. Both transforming growth factor (TGF) alpha and epidermal growth factor (EGF), added to the culture media, stimulated the proliferation of Ishikawa cells in a dose-dependent manner. Anti-TGF-alpha antibody completely eliminated the stimulatory effects of TGF-alpha. Anti-EGF receptor antibody inhibited the proliferation of these cells. The production of TGF-alpha into culture media was 5-40 pg/10 cells/24 h in 9 human endometrial adenocarcinoma cells. Ten nanomoles of estradiol increased the TGF-alpha production of Ishikawa cells by approximately 2.5-fold of the control level. In contrast, the production of TGF-alpha in hormone-unresponsive HEC-50 cels was not influenced by estradiol. C-erbB-2 oncoprotein expression of human endometrial adenocarcinoma cells, detected by both immunocytochemical staining and Western blot analysis, was associated with the tumor grade of the original tumor tissues. Ten nanomoles of estradiol clearly increased the c-erbB-2 oncoprotein levels at an optimal incubation period of 72 h, whereas estradiol did not affect the expression in HEC-50 cells. PMID- 9852401 TI - Requirement of estrogen receptor expression and function for [12Val] K-Ras mediated NIH3T3 cell transformation. AB - We investigated the biological significance of estrogen receptors (ERs) in NIH3T3 cell transformation by the [12Val] K-Ras mutant. This mutant enhanced the steady level and transcriptional activity of ER. Coexpression of the progesterone receptor with mutant K-Ras led to suppression of tumorigenicity and inhibition of the activation of ER. The antisense oligomers complementary to the ER suppressed proliferation and transformed phenotypes of K12V cells. These observations support the importance of ER in Ras-mediated cell transformation. PMID- 9852402 TI - Estrogen dependency in uterine endometrial cancers. AB - Estrogen dependency in uterine endometrial cancers involves complicated tumor biology. A transformed phenotype of uterine endometrial cancers is supported by estrogen-dependent oncogene (c-Ha-ras, c-fos and c-jun) expressions. The relative overexpression of estrogen receptor exon 5 splicing variant and the damaged expression of progesterone receptor A, which belong to a lack of estrogen dependency, are related to metastatic potential. The estrogen-related metastatic processes, detachment, invasion and angiogenesis in some uterine endometrial cancers can be inhibited by progestins. PMID- 9852403 TI - Creutzfeldt-Jakob disease and the risk from blood or blood products. AB - The occurrence of iatrogenic cases of Creutzfeldt-Jakob disease (CJD) and the isolation of infectivity in some laboratory transmission studies in transmissible spongiform encephalopathies raises the possibility that CJD might be accidentally transmissible through blood or blood products. Epidemiological evidence, although not conclusive, does not suggest that classical CJD is transmitted through this route. However, new variant CJD (nvCJD) might pose greater risks of accidental transmission of infection and mechanisms to reduce the theoretical risk are under consideration. The theoretical risks from CJD and nvCJD must be balanced against the established therapeutic benefits of blood and blood products. PMID- 9852404 TI - Comparison of filtration properties of hepatitis B virus, hepatitis C virus and simian virus 40 using a polyvinylidene fluoride membrane filter. AB - BACKGROUND AND OBJECTIVES: We examined the ability of a modified polyvinylidene fluoride membrane filter to remove blood-borne and surrogate viruses. MATERIALS AND METHODS: Phages PR772 and PP7, hepatitis B virus (HBV), hepatitis C virus and simian virus 40 (SV40) were spiked in minimum essential media with 10% fetal calf serum and the concentration of these viruses compared before and after filtration by either plaque assay or polymerase chain reaction. RESULTS: Viruses >50 nm were removed to below detection limits (>10(6) logs) for all filters tested. A 5-log reduction of HBV (42 nm) and 2- to 3-log reduction of HCV (30-65 nm) was observed. CONCLUSION: A predictable size-based removal of viral agents was observed. The results also suggest the possible utility of SV40 as a surrogate to HBV for membrane filter challenge studies. PMID- 9852405 TI - GB virus C/hepatitis G virus and intravenous immunoglobulins. AB - BACKGROUND AND OBJECTIVES: Different intravenous immunoglobulins (IVIGs) were found to be GB virus C/hepatitis G virus polymerase chain reaction (GBV-C/HGV PCR)-positive. The potential transmission of this virus to recipients by a PCR positive IVIG batch was investigated. MATERIALS AND METHODS: Polyclonal IVIGs of different manufacturers and with different virus inactivation procedures were analyzed by GBV-C/HGV-PCR and anti-E2-ELISA. Follow-up sera of 13 participants of a clinical trial performed with a GBV-C/HGV-PCR-positive batch were retrospectively investigated for GBV-C/HGV seroconversion (specific antibodies, viral RNA). RESULTS: Four out of ten IVIGs analysed by GBV-C/HGV-PCR were - at least for some batches - virus genome-positive. Virus inactivation by solvent/detergent treatment resulted in GBV-C/HGV-PCR-negative products. GBV C/HGV-specific antibodies were detectable in all IVIGs analyzed. There was no transmission of GBV-C/HGV observed when recipients of a large amount of a GBV C/HGV-PCR-positive batch were analyzed by an antibody test and specific PCR. CONCLUSIONS: Despite PCR positivity of an IVIG preparation no transmission of GBV C/HGV to recipients was observed. Possible explanations are a sufficient virus inactivation procedure and/or presence of specific antibodies in the final products. PMID- 9852406 TI - Comparison of seroprevalences of human herpesvirus-6 and -7 in healthy blood donors from nine countries. AB - BACKGROUND AND OBJECTIVES: The purpose of the study was to register antibody prevalences of HHV-7 in various locations of the world in comparison to the closely related HHV-6. MATERIALS AND METHODS: Sera of healthy blood donors from nine countries in five continents were titered by indirect immunofluorescent assays using HHV-6 infected HSB2 and HHV-7 infected SupT1 cells. RESULTS: Antibody prevalence for HHV-7 is high (75-98%) in practically all countries except for Northern Japan (44%), with no simple correlation to elevated HHV-6 antibody titers. There were regions of low, intermediate and high mean antibody titers against HHV-7 such as 78.5-91.3 for Belgium, Israel, Japan, USA and Australia, 175.4-182.6 for Mexico and Cologne/Germany, and 389.2 for South Africa for which geographic characteristics may be responsible. CONCLUSION: HHV-7, similar to HHV-6, is a widespread human herpesvirus with elevated antibody titers in the healthy human population essentially everywhere. The data warrant further studies to evaluate its possible pathologic potential, preferentially in persons with defective immune responses. PMID- 9852407 TI - Increased lipid peroxidation of erythrocytes in blood stored in polyvinyl chloride blood storage bags plasticized with di-[2-ethyl hexyl] phthalate and effect of antioxidants. AB - BACKGROUND AND OBJECTIVES: Previous work in this laboratory has shown significant decrease in vitamin E in erythrocytes in blood stored in polyvinyl chloride (PVC) bags plasticized with di-[2-ethyl hexyl] phthalate (DEHP), and in erythrocytes incubated in vitro with DEHP. Since vitamin E is a major antioxidant, a study was carried out to find out whether this decrease observed in vitamin E has an effect on lipid peroxidation in blood stored in DEHP-plasticized PVC blood bags. MATERIALS AND METHODS: Blood was collected in Penpol blood storage bags (which is a DEHP-plasticized PVC bag) and parameters of lipid peroxidation, i.e. activity of superoxide dismutase (SOD), catalase, glutathione peroxidase, glutathione reductase, concentration of malondialdehyde (MDA), conjugated dienes, hydroperoxides, glutathione and vitamin E studied in erythrocytes after various periods of storage as compared to glass bottles. Erythrocytes were also incubated in vitro with DEHP with and without vitamin E, and changes in lipid peroxidation studied. RESULTS: Blood stored in Penpol bags showed increased lipid peroxidation in erythrocytes as compared to that stored in glass bottles, as is evident from a greater increase in MDA and a greater decrease in glutathione and a significant decrease in vitamin E. The addition of vitamin E decreased the formation of MDA and conjugated dienes and prevented the decrease in vitamin E. However in spite of increased lipid peroxidation in the presence of DEHP, the release of K+ and hemoglobin from erythrocytes was lower. When there was an increase in DEHP taken up by erythrocytes, there was a corresponding decrease in vitamin E. More important, whenever there was an increase in vitamin E in erythrocytes (when RBCs in the presence of DEHP were incubated with vitamin E), there was a progressive decrease in DEHP. CONCLUSION: DEHP caused increased lipid peroxidation in erythrocytes. At the same time, it decreased the release of K+ and hemoglobin from erythrocytes. It is possible that the stabilizing effect of DEHP on the erythrocyte membrane may offset the detrimental effects of the increased lipid peroxidation it causes. PMID- 9852409 TI - Evaluation of in vivo and in vitro quality of apheresis-collected RBC stored for 42 days. AB - BACKGROUND AND OBJECTIVES: New technological developments make it possible to collect red blood cells (RBCs) by apheresis, which allows for better product consistency and has the potential for improved RBC quality. The purpose of these studies was to evaluate the quality and consistency of units of RBCs collected by apheresis using the MCS+(R) machine (Haemonetics Corp., Braintree, Mass., USA). MATERIALS AND METHODS: Two studies were performed. In study 1 (n = 10), using containers and CP2D/AS-3 solutions from Medsep Corp. (Covina, Calif. USA), one unit apheresis RBCs were compared to manually collected RBCs in a random crossover design. In study 2 (n = 12), 6 subjects had one unit collected, while the remaining 6 subjects had two units of RBCs collected with comparison to previously manually collected RBCs from the same donors. Haemonetics containers and solutions were used in study 2. RESULTS: Low RBC volume variability was found for the apheresis collections with a standard deviation of only 6 ml difference between actual and target volumes. Combining the data from the two studies (n = 21 pairs), at 42 days of storage, the apheresis units showed slightly lower hemolysis (0.44+/-0.26 vs. 0.61+/-0.50%), lower supernatant potassium levels (50+/-3 vs. 53+/-3 mEq/l), and improved tolerance to osmotic shock (47+/-3 vs. 49+/-3%) as compared to manual units (p < 0.05). There was no statistically significant difference in RBC ATP (3.0+/-0.6 vs. 2.9+/-0.5 micromol/g Hb) or in 24-hour percent recoveries (81+/-6 for apheresis vs. 81+/-4% for apheresis red cells). Apheresis RBC quality was not affected by the manufacturer (Haemonetics vs. Medsep) of solutions and containers. CONCLUSIONS: RBC units collected by apheresis demonstrated low variability in volume of RBC mass collected, and showed similar RBC properties as compared to manually collected RBCs after processing and after 42 days of storage. PMID- 9852408 TI - Erythrocytes and proinflammatory mediators in wound drainage. AB - BACKGROUND AND OBJECTIVES: Retransfusion of shed blood collected after operation has become popular, but recent reports of side effects led to a search for possible causes. MATERIALS AND METHODS: In a randomized study of 28 patients undergoing total hip arthroplasty, shed blood was collected in Solcotrans, Orth Evac, and ordinary Redon drainage. Osmotic fragility was measured and electron microscopic pictures of erythrocytes from selective samples were taken. Serotonin, prostaglandin E2 (PGE2), and histamine were measured with enzyme linked immunosorbent assays. RESULTS: Higher osmotic fragility of erythrocytes collected with Solcotrans appeared to be due to ACD which was used only with that system. Serotonin concentrations did not differ significantly. However, there was a great increase in histamine (Solcotrans 477.7, Orth-Evac 344.0, Redon drainage 453.1 nmol/ml) and PGE2 (Solcotrans 1,908.3, Orth-Evac 1,225.0, Redon drainage 2,666.7 microgram/ml) in shed blood compared with venous blood (histamine 9.5 nmol/l, PGE2 4.2 microgram/ml). CONCLUSION: Unwashed wound drainage blood collected after operation contains levels of proinflammatory mediators that can account for the reported side effects. PMID- 9852410 TI - Platelets stored in a new-generation container differences between plasma and platelet additive solution II. AB - BACKGROUND AND OBJECTIVES: In order to preserve platelet concentrates (PCs) with high yields, a new polyolefin platelet storage container (PL 2410, 1.3L, Baxter, La Chatre, France) with increased gas permeability in combination with a larger surface area has been developed. The storage capacity was studied with platelets in plasma and platelets additive solution. MATERIALS AND METHODS: Platelet concentrate pools (PCs) of different yields suspended in either plasma (PCs-PL; n = 30) or PAS II (PCs-PAS; n = 37) were prepared. For preparation of PCs with a low, intermediate and high number of platelets 3, 5 and 6 buffy coat (BCs) were pooled with different volumes of plasma and 5 and 6 BCs were pooled with different volumes of PAS II, in order to obtain PCs of equal volumes comparable with routine conditions. All PCs were stored on a flatbed shaker at 22+/-2 degrees C and evaluated on days 1, 2, 5 and 7 by determining platelet and white cell counts, pH (37 degrees C), pO2, pCO2 and swirling score. RESULTS: Platelet yields ranged from 1.5 up to 5.5 x 10(11) platelets/U. On day 7 all PCs-PL (n = 4) with platelet yields above 4.5 x 10(11)/U had a pH value below 6.8 (range 5.91 6.79). While 7 of 8 PCs-PAS units with platelet yields above 4. 0 x 10(11)/U showed a pH value below 6.8 (range 6.31-6.70). CONCLUSION: During 7 days of storage in a new 1.3-liter platelet container, the pH was maintained above 6.8 in PCs in plasma with a yield between 1.5- and 4.5 x 10(11)/U; when PAS II was used, the maximum platelet yield allowed for comparable pH maintenance was somewhat lower (4.0 x 10(11)/U). PMID- 9852411 TI - Autologous platelet collection and storage to support thrombocytopenia in patients undergoing high-dose chemotherapy and circulating progenitor cell transplantation for high-risk breast cancer. AB - OBJECTIVES: The use of circulating progenitor cell support following high-dose chemotherapy for malignancies decreases but does not entirely abolish platelet transfusion requirement. We investigated the feasibility of supporting the posttransplant thrombocytopenic phase exclusively with autologous platelets collected by apheresis and cryopreserved. METHODS: 25 patients underwent plateletpheresis during the platelet rebound occurring after high-dose cyclophosphamide. Autologous platelets were cryopreserved in 5% dimethylsulfoxide, thawed and transfused during the aplastic phase after the myeloablative regimen whenever clinically required. RESULTS: A single plateletpheresis was carried out in all patients, allowing the harvest of a platelet concentrate with a mean value of 7.7 x 10(11) platelets. No significant procedure- or transfusion-related side effects were recorded. Mean platelet recovery after freezing and thawing was 63% and the mean number of platelet reinfused was 4.8 x 10(11); 23 of 25 patients were fully supported with autologous platelets. CONCLUSION: Plateletpheresis performed in our selected group of patients was found to be a safe and effective procedure to collect large amounts of autologous platelets; the numbers obtained proved to be sufficient for the transfusion demand of almost all patients. PMID- 9852412 TI - Two examples of an inseparable antibody that reacts equally well with DW+ and Rh32+ red blood cells. AB - BACKGROUND AND OBJECTIVES: Anti-DW is a rare specificity that detects an antigen on DVa red blood cells (RBCs). Some anti-DW contain an inseparable component that cross-reacts weakly with RBCs expressing the low-incidence Rh antigen Rh32. RH32 is expressed by RBCs with either the R=Nor the DBT phenotype. CASE REPORT: We describe here an antibody found in the serum of 2 patients that reacts equally well with RBCs possessing either DVa, R=N, or DBT phenotypes. The reactivity for DW and Rh32 antigens could not be separated by adsorption onto and elution from DW+Rh32- or from DW-Rh32+ RBCs. CONCLUSIONS: We suggest that amino acids encoded by nucleotides at the junction of exon 4 of RHD to exon 5 of RHCE may induce a conformation that is recognized by these equally reactive inseparable antibodies. Until such time that the epitope recognized by these antibodies is defined, we recommend use of the descriptive name anti-DW/Rh32. PMID- 9852413 TI - A flow-cytometric equivalent of the Kleihauer test. AB - BACKGROUND AND OBJECTIVES: The Kleihauer slide test is in general use to screen obstetric patients for possible fetomaternal haemorrhage. Since 1993, Rh(D) negative patients have been tested in our laboratory by a flow-cytometric method detecting Rh(D)-positive fetal cells, a method which offers improved sensitivity and accuracy. We report another flow-cytometric method of broader application which quantitates cells according to haemoglobin F (HbF) content. MATERIALS AND METHODS: The red cells are fixed with glutaraldehyde and permeabilized by exposure to Triton X-100. A polyclonal sheep antibody to HbF is incubated with the cells followed by a fluorescein-labelled anti-sheep antibody. RESULTS: Quantitation of the percentage of fetal cells following a FMH can be achieved irrespective of the blood groups of either mother or infant, and the presence of maternal F cells need not interfere since the intensity of staining is usually less than that of fetal cells. Two of 19 transfusion-dependent patients with beta thalassaemia have been found to have red cells indistinguishable from fetal cells on the basis of HbF content, but these patients also have been found to give positive results by the Kleihauer test. CONCLUSIONS: The flow-cytometric method may serve to replace the traditional Kleihauer test since it appears to offer improved accuracy and objectivity. PMID- 9852414 TI - Three cases of platelet alloimmunisation associated with the presence of a novel platelet-specific antibody. AB - BACKGROUND AND OBJECTIVES: In three cases of platelet alloimmunisation, a platelet-specific alloantibody was detected which could not be classified within the known human platelet alloantigen or HLA systems. The first case was of a family in which two siblings suffered neonatal alloimmune thrombocytopenia at birth. In the second case, the newborn was suffering from phocomelia with hypoplastic thrombocytopenia. The third case was a male who became refractory to transfusions of HLA-matched platelets after a related bone marrow transplantation. MATERIALS AND METHODS: The serum samples were investigated by: enzyme-linked immunosorbent assay, platelet suspension immunofluorescence test (PSIFT), monoclonal antibody immobilisation of platelet antigens assay (MAIPA), and by the lymphocytotoxicity test. RESULTS: The antibody gave positive reactions with 26% of normal donor platelets. Surprisingly, no platelet-specific antibody was detected by PSIFT or by MAIPA and there was no evidence found to support classifying the antibody within the HLA system. CONCLUSION: The reactivity pattern of the antibody detected and the clinical presentation of the three cases described, strongly suggest the presence of an additional platelet-specific alloantigen system. PMID- 9852415 TI - NA1/NA2 antisera inhibit FcgammaRI- but not FcgammaRII- mediated phagocytosis. AB - BACKGROUND AND OBJECTIVES: Alloantibodies against the granulocyte-specific NA antigens play an important role in alloimmune neonatal neutropenia. As the NA system is located on the FcgammaRIIIb, the influence of NA-specific antibodies on granulocyte function is of special interest. MATERIALS AND METHODS: We tested alloantisera specific for NA1 and NA2 for their ability to influence FcgammaR mediated phagocytosis of polymorphonuclear neutrophils by use of different FcgammaR-specific targets. Red blood cells coated with human IgG anti-D served as specific targets for FcgammaRI-mediated phagocytosis while mouse IgG1 anti glycophorin A was used to coat red blood cells (RBCs) to obtain FcgammaRII specific targets. To test for a hypothetical induction of phagocytosis by FcgammaRIIIb we used D-- RBCs coated with human monoclonal anti-D as target cells for unprimed neutrophils. RESULTS: Granulocyte phagocytosis was directly induced by FcgammaRI and FcgammaRII but not by FcgammaRIIIb. NA1 alloantisera significantly inhibited FcgammaRI-mediated phagocytosis of IFN-gamma-stimulated neutrophils if the corresponding antigen was expressed. Conversely, NA2 alloantisera inhibited FcgammaRI-mediated phagocytosis in NA2-positive individuals. There was no effect of NA1- and NA2-specific alloantibodies on FcgammaRII-mediated phagocytosis. CONCLUSION: NA-specific alloantisera inhibit the FcgammaRI-induced phagocytosis in primed neutrophils, but they do not significantly inhibit their FcgammaRIIa-specific phagocytosis of mIgG1-coated RBCs. PMID- 9852416 TI - Rapid detection of genetic mutations associated with haemochromatosis. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to establish a rapid method suitable for large-scale population screening, including blood donors, for the detection of two genetic mutations at codons 63 and 282 on the HFE gene that are associated with haemochromatosis. MATERIALS AND METHODS: A method using the polymerase chain reaction with sequence-specific primers (PCR-SSP) was designed and tested using a panel of 185 individuals previously typed for HFE mutations by PCR-RFLP. RESULTS: The PCR-SSP method detected the two mutations showing complete agreement with the genotypes obtained by PCR-RFLP. Three HFE alleles, termed HFE 1, -2, and -3, were identified. CONCLUSIONS: This PCR-SSP method allows efficient HFE genotyping for large numbers of individuals. PMID- 9852417 TI - Autoimmune haemolytic anaemia induced by propyphenazone. PMID- 9852418 TI - Follow-up to HBV and HCV carriers in a blood center. PMID- 9852420 TI - Re: Grand rounds #48. A case of residual superior oblique palsy and contralateral inferior oblique palsy (surgically induced) after bilateral surgery for presumed bilateral superior oblique palsy. PMID- 9852419 TI - Re: Caldeira JAF: Stage I intraoperative adjustment of eye muscle surgery forexodeviations based on eye position during general anesthesia: a prospective study of 35 patients. PMID- 9852421 TI - Botulinum toxin chemodenervation of the inferior oblique muscle for chronic and acute IV nerve palsies: results in 15 cases. PMID- 9852422 TI - Tchebysheff! Yet another reason you should NOT use "statistical significance" = a probability < or = .05. PMID- 9852423 TI - Downshoot in infra-adduction following selected superior oblique surgical weakening procedures for A-pattern strabismus. AB - PURPOSE: To determine the incidence of postoperative persistent or recurrent, true or pseudo, superior oblique muscle overaction (SOOA) following treatment of A-pattern strabismus by a selective surgical approach; and to seek its cause. SUBJECTS AND METHODS: Retrospective analysis of the records of 104 consecutive patients with A-pattern strabismus and SOOA treated, with 4 different superior oblique (SO) weakening procedures, selected according to the amount of preoperative anisotropia (in order, smallest to largest): posterior (partial) tenectomy, scleral disinsertion, tenectomy at the insertion and recession with posterior transposition. After exclusions for cause, there were 93 patients who received the same procedure in both eyes. The mean preoperative A anisotropia was 18.30delta+/-8.05delta (range 8delta to 50delta). The mean preoperative SOOA was +2.06 (range +1 to +4). RESULTS: The mean correction obtained by surgery was 16.20delta+/-8.7delta (88.5% of the preoperative deviation). The mean reduction of SOOA was -1.89 (-91. 7% of the preoperative SOOA, range -75.8% to - 103.1%). During postoperative followup, a pattern of persistence of downshoot in the infra adducted position of gaze (true/pseudo SOOA) was found in 37. 6% of the cases (28% following tenectomy, 30.7% after recession, 40. 4% after posterior tenectomy and 62.5% after disinsertion). A complication, Superior Oblique Inclusion Syndrome was discovered in two cases. CONCLUSIONS: Persistence or recurrence of postoperative downshoot in the infra-adducted position is a common finding following any of the SO weakening procedures analyzed in this study. Its occurrence does not seem to be related either to the amount of preoperative SOOA and/or A anisotropia, nor to the amount of A anisotropia correction obtained. Relatively uncontrolled procedures, such as disinsertion, showed the highest incidence of this sequel of surgery. PMID- 9852424 TI - A protocol for extraocular muscle surgery after orbital floor fracture ("blow out") AB - PURPOSE: If, after primary repair of an orbital fracture by maxillo-facial surgeons, diplopia persists, extraocular muscle surgery may be necessary. It was the purpose of this study to examine proposed surgical treatment in these cases to determine their efficacy. SUBJECTS AND METHODS: We analyzed, retrospectively, the files of 14 patients who were treated in our strabismological department for persistent diplopia caused by injury to the extraocular muscles and/or to the surrounding structures. The aim of our treatment protocol was to restore comfortable binocular single vision in primary position and downgaze. The selection of the surgical procedure depended on the deviation in primary position and on the degree of ocular muscle imbalance. Surgery consisted of recession of the inferior rectus muscle of the affected eye in 4 cases, resection of the inferior rectus muscle of the affected eye in 4 other cases, and large recessions of the vertical muscles of the contralateral normal eye in 6 cases. RESULTS: In all 14 patients, we obtained the desired comfortable field of binocular single vision, considered "good" (20 degrees up to 30 degrees downgaze) or "satisfactory" (a useful field). In 11 cases this was achieved in one procedure. Two patients needed a reintervention because of initial overcorrection, and one patient for an initial undercorrection. All 14 patients had a "good" or "satisfactory" final result (useful binocular single vision). CONCLUSIONS: When extraocular muscle surgery is necessary, we recommend in patients with limited forced elevation, recession of the taut rectus inferior muscle; in patients with the clinical characteristics of an inferior rectus palsy, a resection of this muscle; and in cases with a normal or only slightly limited forced duction test, weakening the contralateral synergists. PMID- 9852425 TI - Grand rounds #49: A case of bilateral asymmetric Duane syndrome with fixation with the more affected eye. PMID- 9852426 TI - Diagnosis and treatment of isolated inferior oblique muscle palsy: a report of seven cases. AB - PURPOSE: To review and report our experience in attempting to surgically eliminate the vertical deviations and the anomalous head posture in patients with inferior oblique palsy (IOP) and to add to the literature another etiology for IOP: denervation/myectomy of that muscle. SUBJECTS AND METHODS: Seven patients demonstrating findings characteristic of an isolated IOP are reported. Six were unilateral and one was bilateral. All met the Bielschowsky/Parks' Three Step Test criteria to identify an isolated IOP. Six patients had surgery that consisted of weakening the superior oblique (SO) muscle of the affected eye alone or in combination with superior rectus recession of the fellow eye. RESULTS: All patients were ultimately successfully alleviated of their vertical deviations and anomalous head postures. (Two patients developed an iatrogenic superior oblique palsy (SOP) which responded well to additional surgery.) CONCLUSIONS: The treatment of choice for IOP is tenotomy or tenectomy of the ipsilateral superior oblique muscle alone or in combination with recession of the contralateral superior rectus muscle. PMID- 9852427 TI - Case report: benign recurrent abducens nerve palsy in children: another triggering factor: impetigo. PMID- 9852428 TI - 'Managed' care somewhat 'mangled'? PMID- 9852429 TI - Re: van Eeckhoutte L. et al. A protocol for extraocular muscle surgery afterorbital floor fracture ("blow-out") PMID- 9852431 TI - Results of large (8-9 mm) bilateral lateral rectus muscle recessions for exotropia. AB - PURPOSE: Commonly recommended maximum lateral rectus muscle recession for exotropia ranges from as little as 7 mm to as much as 11 mm. Published studies on recessions of 8 mm or more are scarce. Many ophthalmic surgeons still avoid recessions larger than 7 mm because of the fear of creating abduction deficits and surgical overcorrections. Therefore, we sought to determine the incidence of clinically detectable abduction deficits and their correlation with early surgical overcorrection in exotropic patients who had undergone large 8 to 9 mm lateral rectus muscle recessions. SUBJECTS AND METHODS: A retrospective chart review identified 30 patients with exotropia ranging from 35delta to 65delta who were treated with 8 to 9 mm recessions of both lateral rectus muscles. Patients with concurrent oblique muscle surgery were included. Mean followup time was 15 months (range 3-30 mos). RESULTS: Twenty-four patients (80%) required only one operation. The remaining 6 patients (20%) required a second operation, four for overcorrection and two for undercorrection. Nine patients (30%) had mild but persistent abduction deficits postoperatively. However, abduction deficits were not predictive of poor outcome (p=0.959). Other variables that did not significantly affect outcome included age (p=0.894), systemic anomalies (p=0.127), size of preoperative exotropia (p=0.987) and amount of rectus muscle recession (p=0.480). However, concurrent oblique muscle surgery was associated with a higher risk of a poor result. CONCLUSIONS: 8-9 mm lateral rectus recessions are not associated with a poor outcome, except in association with concurrent oblique muscle surgery. PMID- 9852432 TI - Video loop MRI of ocular motility: a new technique: turbo rare sequence at 2 T for the study of horizontal gaze. AB - PURPOSE: To improve existing MRI (Magnetic Resonance Imaging) methods of assessing ocular motility in horizontal gaze by using a multi-static fast imaging T2W MRI sequence, preventing patient head motion and convergence of the eyes during the study for the diagnosis and management of complex paralytic strabismus. SUBJECTS AND METHODS: A turbo-RARE image of the orbits was acquired in a set plane each time the patient fixed on a virtual image of one of 15 LEDs (Light Emitting Diodes). Positioning and securing the head was by a security cushion with an 8 degree tilt of the 25 cm quadrature head coil. After completion of the study (5 minutes) the MRI images were transferred to a work station and on to a magnetic-video tape to be replayed in cine mode on a PC using standard software. The qualitative analysis was completed by the study of quantitative parameters: visual axis, minor axis of the lens, the lines and angles of the horizontal rectus muscles with the optic nerve and the visual axis. We used digital image analysis to measure the cross-sectional areas of these muscles and also to determine muscle volumes. Twenty normal volunteers and 3 cases of oculomotor palsies were so studied. RESULTS: The experimental setup allowed us to obtain: 1) rapid (14 s) and high resolution images (256 x 196 x 3 mm); 2) which were free artifacts; 3) with no detectable eye convergence; 4) in which the lateral and medial rectus muscles and anatomical details of the orbit were clearly depicted; 5) from which additional functional information was obtained from the cine loop analysis; 6) wherein the visual axes did not converge during horizontal versions; 7) which permitted horizontal rectus muscle angles to be followed during contraction and relaxation; and 8) allowed the cross-sectional area of the four rectus muscles to be compared during contraction and relaxation (difference of 0.2 cm2). CONCLUSIONS: This method will provide useful functional quantitative information in the evaluation of ocular motility disorders. The procedure is readily clinically applicable PMID- 9852433 TI - Grand rounds #50: A case of a lost medial rectus muscle in an eye in which the inferior rectus muscle "pulled in two". PMID- 9852435 TI - Major review: ocular sighting dominance: a review and a study of athletic proficiency and eye-hand dominance in a collegiate baseball team. AB - PURPOSE: REVIEW: Laterality preference in sensory and motor functions of symmetrically disposed organisms have been studied for centuries. The relation between handedness and the eyes and vision (ocular sighting dominance) has been a focal point despite their physiologic dissimilarity. STUDY: To examine a college varsity baseball team for handedness and ocular sighting dominance to determine if their patterns of eye-hand dominance differed from the normal population and/or contributed to their individual relative success compared to their peers. Specifically: whether crossed eye- hand dominance favors the batter and uncrossed eye-hand dominance favors the pitcher. SUBJECTS AND METHODS: Twenty five UF varsity players were examined. All were male. Their visual acuity, stereoscopic vision, ocular motility and ocular sighting dominance were determined, the last by a pointing test which allowed the diagnosis of a central form of ocular dominance, but was not per se affected by handedness. Handedness was determined by preferred arm for throwing or hitting. No subject was ambidextrous. A control population was established consisting of the first 100 consecutive adults seen by the first author in the UF Eye Center with 20/20 vision O.U. and a normal eye exam. RESULTS: The control group displayed eye-hand dominance patterns similar to those previously reported in the literature for the general population. In the experimental group of baseball players, the incidence of conventionally predominant (in normals) ipsilateral or uncrossed eye-hand dominance was much lower (39%) than the normal control population (65%). The incidence of contralateral or crossed eye- hand dominance was 35%, twice that of the normal control population (18%) (p<0.01). The incidence of central ocular dominance with right or left handedness was 26% or 50% higher than a normal control population (17%) (p<0.25). With regard to individual performance, those players with central ocular dominance, whether right or left handed, were the most successful players in both pitching and batting. Pitchers who were uncrossed eye-hand dominant were distinctly more successful than crossed. Batters who were crossed eye-hand dominant were slightly more successful than uncrossed. CONCLUSIONS: The pattern of eye-hand dominance appears related to athletic proficiency for baseball. Warning: Note well: This information may be useful in career guidance but does not justify or medically indicate attempts to alter ocular sighting dominance or eye-hand dominance patterns as these are determined probably genetically or at such an early age that they cannot be successfully altered later. Attempts to so alter them are historically fraught with irremediable psychological or physical injury to the subject, including permanent incapacitating double vision. PMID- 9852437 TI - Respecting the superior oblique tendon. PMID- 9852436 TI - Ten days and nine nights in Moldova. PMID- 9852438 TI - Blue filter amblyopia treatment protocol for strabismic amblyopia: a prospective comparative study of 50 cases. AB - PURPOSE: Previous studies of monochromatic visual evoked potentials confirm the strong suppression of (the cortical representation of) paracentral retinal areas of functionally amblyopic eyes, by a flat response to a blue stimulus. A clinical trial stimulating these areas with blue light was encouraging, and justified a prospective comparison of this treatment with conventional classic treatment. SUBJECTS AND METHODS: In 50 strabismic children with amblyopia, the blue filter treatment protocol (flash stimulation with, and the wearing of, a blue filter during occlusion of the better eye for one hour daily) was compared with the classical treatment (full time total occlusion by patch) in a prospective matched and randomized study. Patients 3 to 7 years old without previous treatment and a visual acuity up to 0.3 were admitted to the study. Visual acuity and fixation behavior were used as the parameters of comparison. Visual Evoked Potentials by monochromatic flashes were also studied. Results of treatment were compared after 6 months. RESULTS: The visual acuity outcome for the blue filter treatment was "statistically significantly" better (p=0.005). The greatest improvement was seen in the subgroup of children with eccentric fixation (p=0.01). Fixation behavior also showed a better outcome from the blue filter treatment (p=0.05) favoring especially children between 3 and 5 years. In children of this age with a visual acuity better than 0.1 we found a very "statistically significant" difference between the two treatments (p=0.004). In children 3 to 5 years old with poorer visual acuity we also found a "statistically significant" difference in the two treatments (p=0.04). The interocular difference of amplitude on the Visual Evoked Potentials also demonstrated more improvement in children treated with the blue filter. This treatment improved especially the cortical response to blue flash stimulation, correlating to paracentral retinal areas. CONCLUSIONS: The blue filter treatment protocol provided better results for treating amblyopia than the conventional classic occlusion treatment method. We propose that stimulation of these paracentral retinal areas triggers a better disinhibition of a functionally amblyopic eye. PMID- 9852439 TI - Long term outcome of treated congenital/infantile esotropia: does early surgical binocular alignment restoring (subnormal) binocular vision guarantee stability? AB - PURPOSE: To answer this important outcome question. SUBJECTS AND METHODS: Retrospective review of a series of 60 qualified cases of congenital/infantile esotropia surgically binocularly "aligned" (= orthotropia +/- 8 PD) before 30 months of age and showing at one postoperative examination date at least some degree of (subnormal) binocular vision. Every patient was monitored over a very long postoperative followup period (mean 14.7 +/- 3.7 years). RESULTS: Early binocular alignment with some binocular cooperation does not guarantee a stable outcome. However, even though the final percentage of patients losing "alignment" is similar to that observed in other non-qualified series of "aligned" congenital/infantile esotropias, the loss of "alignment" takes place at a much later age, if some binocular cooperation was present at some time in the past. CONCLUSIONS: For congenital/infantile esotropia, achieving, at an early age, binocular "alignment" with some degree of binocular vision delays the loss of "alignment" and seems to reduce the chance of developing oblique muscle overactions. PMID- 9852440 TI - A biomechanical model of congenital/infantile esotropia and its treatment. AB - PURPOSE: To evaluate the ability of a computer assisted model of ocular motility, to reproduce this syndrome using known ocular anatomic biomechanical muscular parameters, and to hypothesize pathogenetic mechanisms deduced from clinical and laboratory experiences. METHODS: The manipulation of anatomic, elastic, contractile and innervational parameters available in the computer software program Orbit 1.6, were used to model: a) anatomic ocular and orbital parameters found in the 12 months old infant; b) static (biomechanical) changes measured intraoperatively in the extraocular muscles (medial and lateral rectus muscles) of infantile esotropes; and c) dynamic components of the horizontal deviation, including crossed fixation and bilateral limitation of abduction. Bilateral medial rectus recession surgery was also simulated on the completed model by moving backwards the insertions of both medial rectus muscles. RESULTS: Small, fairly comitant, esotropic deviation were simulated using real data of the mechanical status of medial and lateral rectus muscles collected intraoperatively (length-tension measurements). The more typical large esotropic deviations with bilateral limitation of abduction of the congenital/infantile esotropia syndrome were only obtained when subtle modifications of the medial and lateral rectus muscles' thresholds to innervation were added to the anatomical-mechanical model. An orthotropic primary position binocular alignment with full rotations was obtained in this model following standard bilateral medial rectus recessions, showing surgical dose/response figures close to those usually found in the treatment of real cases of congenital/infantile esotropia. CONCLUSIONS: This biomechanical computer-assisted model may provide a very useful tool not only to test new or different surgical procedures and dosages but also to explore different pathogenetic mechanisms. PMID- 9852441 TI - Synostotic plagiocephaly causing pseudoparalysis of the superior oblique and ocular torticollis: report of a case with unique sensory findings. AB - PURPOSE: To report a case in which strabismus surgery did not eliminate the abnormal head posture although it did improve the vertical strabismus; and to consider the causes of this situation. CASE REPORT: The patient had a right eso hypertropia and ocular torticollis to the left shoulder as a result of a right superior oblique mechanical deficiency, secondary to her craniosynostosis. A myectomy of the right inferior oblique muscle and a tucking procedure of the right superior oblique muscle were performed. Postoperatively, the abnormal head posture persisted, although strabismus surgery improved the vertical deviation. However, improvement of the abnormal head posture upon patching the deviated eye only indicated that the origin of the persistent abnormal head posture must be ocular. Further tests revealed that an anomalous retinal correspondence (ARC) with eccentric fixation was responsible. CONCLUSIONS: In patients with ocular torticollis, ARC with eccentric fixation must be ruled out prior to surgery as these may negatively affect the outcome. PMID- 9852442 TI - Outcome of surgical management of superior oblique palsy: a study of 123 cases. AB - PURPOSE: To determine the outcome of the surgical management of superior oblique palsy at our institution. SUBJECTS AND METHODS: Retrospective review of 123 patients who underwent surgical correction of superior oblique paresis at Bascom Palmer Eye Institute from 1976 to 1996. Subject-Patients: 67% were male and 33% female. The mean age at surgery was 30.5 years (range, 2-78 years). Etiologies of the pareses were trauma (34%), congenital (33%), and acquired/non-traumatic (33%). The mean angle of preoperative vertical deviation in primary gaze was 14.0 delta (range, 0-45 delta). SURGERY: 109/123 (89%) patients underwent single muscle surgery. Of these 109, 57 had single oblique muscle surgery: a superior oblique tuck in 34/57 (60%); an inferior oblique weakening procedure in 22/57 (38%); and a Harada-Ito procedure in 1/57 (2%). The other 14 patients (11%) had bilateral surgery. RESULTS: The final postoperative vertical deviation in primary gaze was < or =3 PD in 60% of patients and < or =7 PD in 80%. The mean change in primary position vertical deviation postoperatively was 10.4 PD for distance and 13.0 PD for near. An "excellent" outcome (final vertical deviation &le3 PD in primary and reading gazes) was achieved most frequently in those patients with congenital pareses and isolated oblique muscle surgery. COMPLICATIONS: Clinically significant Brown's Syndrome occurred in 43/72 (60%) of those cases who had undergone a superior oblique tuck. The incidence of Brown's Syndrome was unrelated to tuck size. Reoperation was three times more likely to be necessary in traumatic cases than in congenital cases (35.0% vs 11.9%, p=0.02). CONCLUSIONS: Based on these results we recommend oblique muscle surgery as the initial procedure to correct superior oblique palsy when appropriate. PMID- 9852443 TI - Grand rounds #52: a case of a residual left exotropia and hypertropia after previous surgery for a third nerve palsy. PMID- 9852444 TI - Bacteremia with esophageal dilation. AB - BACKGROUND: Antibiotic prophylaxis has been recommended for selected patients undergoing esophageal stricture dilation because of a reported high rate of bacteremia. The aim of this study was to determine the rate of bacteremia after esophageal dilatation in a large series and the source of the organisms recovered. METHODS: Blood cultures and oral temperatures were obtained before esophageal dilation and at 5 and 30 minutes after dilation. Dilators were cultured immediately before dilation. Procedural data collected included type of dilation, number of passes, and presence of malignancy. RESULTS: Of 100 procedures in 86 patients undergoing esophageal dilation, 22 (22%) were associated with a positive post-dilation blood culture. Bacteremia was more frequent with dilation of malignant strictures compared with benign strictures (9 of 17 [52.9%] vs. 13 of 83 [15.7%], respectively, p = 0.002) and with passage of multiple dilators compared with passage of a single dilator (16 of 46 [34.8%] versus 6 of 54 [11.1%], respectively, p = 0.007). Bacterial isolates from 22 positive blood cultures matched those from a dilator in only one episode (4.5%). CONCLUSION: The rate of bacteremia after esophageal dilation is 22% and is associated with dilation of malignant strictures or passage of multiple dilators. Organisms cultured from the blood are not transmitted from the dilator. PMID- 9852445 TI - Viridans streptococcal bacteremia after esophageal stricture dilation. AB - BACKGROUND: The incidence of bacteremia with organisms that may cause infective endocarditis after esophageal stricture dilation is unknown. There is disagreement among physicians regarding the need for antibiotic prophylaxis for patients with valvular heart disease undergoing dilation. Our aim was to determine the frequency and duration of bacteremia associated with esophageal stricture dilation. METHODS: Blood cultures were obtained before and after stricture dilation in patients without valvular heart disease and in a control group of patients undergoing upper endoscopy without dilation. RESULTS: A total of 103 patients undergoing dilation and 50 control patients were studied; 22 of 103 patients (21%) undergoing dilation had at least one post-procedure blood culture positive for viridans streptococcus, compared with 1 of 50 (2%) of control patients (p = 0. 001). Blood cultures obtained 1 minute after stricture dilation were positive for viridans streptococcus in 19 of 81 (23%), in 16 of 96 (17%) 5 minutes post-dilation, and in 3 of 63 (5%) 20 to 30 minutes post dilation. Of the 19 patients with viridans streptococcus bacteremia 1 minute after dilation, cultures were still positive in 14 of 19 (74%) at 5 minutes and in 2 of 19 (10%) 20 to 30 minutes post-dilation. CONCLUSIONS: These data support the use of antibiotic prophylaxis before esophageal stricture dilation for patients with valvular heart disease at risk for the development of infective endocarditis. PMID- 9852446 TI - Randomized trial of stent placed above and across the sphincter of Oddi in malignant bile duct obstruction. AB - BACKGROUND: Placement of stents above an intact sphincter of Oddi might prevent migration of bacteria and deposition of organic material into the stent. In patients with malignant obstructive jaundice prolongation of function time of the stent would be expected if it is placed above the sphincter of Oddi. METHODS: Thirty-four patients were randomized to stent placement either above (n = 17) or across (n = 17) the sphincter of Oddi. Straight 10F gauge Teflon stents were used. The patients were evaluated clinically and biochemically at monthly intervals during follow-up. RESULTS: The median stent function time (i.e., the time from insertion of the stent until stent replacement, patient death, or study termination) were 110 days (25th to 75th percentiles, 61 to 320 days) for stents placed above the sphincter of Oddi and 126 days (25th to 75th percentiles, 89 to 175 days) for stents placed across the sphincter of Oddi (nonsignificant [NS]). Stent replacement rates were 58.8% (10 of 17) in patients with stents placed above the sphincter and 29.4% (5 of 17) in patients with stents placed across the sphincter (NS). Significantly more patients in the former group experienced stent migration (9 vs. 2, p = 0.026). The median time from stent insertion until replacement of the stents placed above and across the sphincter of Oddi were 82 days (25th to 75th percentiles, 31 to 185 days) and 89 days (25th to 75th percentiles, 13 to 150 days), respectively (NS). CONCLUSIONS: No significant difference in overall stent performance between the two groups was found, although more stents placed above the sphincter of Oddi migrated. The time until dysfunction of the stent might be increased if migration of stents placed inside the common bile duct could be avoided. PMID- 9852447 TI - Dynamic imaging of the pancreas using real-time endoscopic ultrasonography with secretin stimulation. AB - BACKGROUND: Obstructive disorders of the pancreas, including strictures, stones, sphincter of Oddi dysfunction, and pancreas divisum, are diagnostic and therapeutic challenges. Conventional extracorporeal ultrasound with secretin stimulation has been used as a noninvasive study to detect obstruction and predict outcome of therapy. Inconsistent results have been obtained because of the inherent limitations of standard ultrasonography. The aim of this study was to evaluate the behavior of the main pancreatic duct by endoscopic ultrasonography during secretin stimulation and to diagnose obstructive disorders of the pancreas. METHODS: Secretin-stimulated endoscopic ultrasound (SSEUS, 1 IU/kg secretin) was performed in 20 control subjects (no pancreatic or biliary disease), 40 patients with symptomatic chronic pancreatitis, 40 patients with symptomatic pancreas divisum, 20 patients with suspected sphincter of Oddi dysfunction, and 20 patients with suspected occlusion of pancreatic duct stents. Ductal diameter was measured by endoscopic ultrasonography at baseline and at 1 minute intervals, after administration of secretin, for 15 minutes. A result was determined to be abnormal when a 1 mm or greater dilation of the pancreatic duct was observed from baseline after secretin administration. RESULTS: Of the 40 patients with symptomatic chronic pancreatitis, SSEUS correctly predicted obstructive pathology (stones, strictures) in 12 of 13 patients (92%). Of the 40 patients with symptomatic pancreas divisum, 22 underwent stent therapy (16 of 22 with resolution of symptoms). SSEUS accurately predicted response to stent therapy in 13 patients (81%). Seven of twenty patients with suspected sphincter of Oddi dysfunction had abnormal sphincter manometry. SSEUS accurately predicted sphincter dysfunction in only 4 of 7 patients (57%). Finally, 20 patients with suspected occlusion of pancreatic duct stents were studied. Of the 14 stent occlusions confirmed at ERCP, SSEUS correctly predicted premature occlusion in 12 patients (86%). CONCLUSIONS: SSEUS appears to be a useful diagnostic modality in the evaluation of patients with suspected obstructive disorders of the pancreas and it can predict which patients may respond to endoscopic therapy. PMID- 9852448 TI - Observation of thoracic duct morphology in portal hypertension by endoscopic ultrasound. AB - BACKGROUND: Thoracic duct dilation has been demonstrated in portal hypertension and hepatic cirrhosis by lymphangiography and laparotomy and at autopsy. It is thought to be secondary to increased hepatic lymph flow and has been described in the absence of ascites or esophageal varices. The aim of the present study was to observe thoracic duct morphology by endoscopic ultrasound in various subsets of patients with portal hypertension and hepatic cirrhosis and also to validate existing radiologic/surgical data. METHODS: The thoracic duct of 33 patients with cirrhosis and portal hypertension was studied by endoscopic ultrasound. Patients were divided into four groups: 1, patients with ascites and esophageal varices; 2, esophageal varices without ascites; 3, without esophageal varices or ascites; 4, extrahepatic portal hypertension due to pancreatic malignancy. The thoracic duct diameter was also measured in 14 control subjects (group 5). RESULTS: When the thoracic duct diameter for the five groups was compared with analysis of variance, significance was p < 0.0001; by pairwise comparison, group 1 differed from the other four groups (p < 0.05). Thoracic duct dilation (5.61 mm) was seen in group 1 patients, whereas no dilation was present in groups 2 through 4. Additionally, thoracic duct diameter in 33 portal hypertensive and/or cirrhotic patients was significantly different from that in the 14 control subjects (p = 0. 003). CONCLUSION: The thoracic duct can be reliably identified by EUS in patients with hepatic cirrhosis and portal hypertension. Dilation of the duct is seen only in patients with hepatic cirrhosis, ascites, and esophageal varices. No thoracic duct dilation is present in extrahepatic portal hypertension. Contrary to existing radiologic/surgical data, thoracic duct dilation is not seen in all patients with hepatic cirrhosis and portal hypertension signifying advanced disease. A dilated thoracic duct by endoscopic ultrasound should be considered yet another sign of portal hypertension. PMID- 9852449 TI - Effectiveness of endoscopic sphincterotomy in complicated hepatic hydatid disease. AB - BACKGROUND: The aim of this study was to evaluate the results of endoscopic sphincterotomy in patients with hepatic hydatid cysts that have ruptured into the biliary tract. METHODS: Over a 10-year period, 25 patients (11 men, 14 women, mean age 60 years) underwent treatment; 13 with no prior surgery were treated for biliary obstruction. Postoperative treatment was undertaken in 12 cases because of persistent drainage (8), duct obstruction (3), and postoperative pancreatitis (1). RESULTS: In patients who had not undergone previous surgery, cholangiographic findings were hydatid vesicles in the biliary tract (6), dilation of the biliary tract (3), biliary fistula (2), distal stenosis (1), and purulent bile content after sphincterotomy (1). In all of these cases, sphincterotomy resolved duct obstruction with no complications. All patients with persistent postoperative drainage had a fistula tract between the biliary duct and the cavity, which was resolved by endoscopic treatment in all 8. The 3 patients with postoperative biliary obstruction had hydatid vesicles in the biliary tract, one with a long stenosis resembling sclerosing cholangitis, whereas the patient with postoperative pancreatitis had a distal stenosis. Satisfactory results were obtained with endoscopic sphincterotomy, although 1 patient required a biliary prosthesis. CONCLUSION: Endoscopic sphincterotomy resolves biliary obstruction and postoperative fistulae in most patients with hepatic hydatid cysts that have ruptured into the biliary tract. In some cases, a biliary prosthesis may be required. PMID- 9852450 TI - Randomized controlled studies of injection Gold Probes compared with monotherapies for hemostasis of bleeding canine gastric ulcers. AB - BACKGROUND: There is a significant interest in combination therapy using endoscopic epinephrine injection and thermal coagulation for nonvariceal hemostasis. The purpose of the study was to compare the relative effectiveness, ease of use, and safety of new Injection Gold Probes to other hemostasis techniques in three randomized, controlled laboratory studies of bleeding canine gastric ulcers. METHODS: Fifteen dogs with prehepatic portal hypertension were heparinized and bleeding gastric ulcers were induced with jumbo biopsy forceps. Three different prototypes of Injection Gold Probes were compared with monotherapy (thermal, electrocoagulation, or epinephrine injection alone), control, or combination therapy with separate injector and thermal probes. The treatment times, total number of pulses or injections, volume of epinephrine injected, and ease of applications were recorded. Gastric ulcer size, ulcer healing, and complications were evaluated at 1 and 4 weeks. RESULTS: All endoscopic treatments were effective for acute hemostasis compared with control. Thermal coagulation alone was the fastest treatment to perform. The performance of the first Injection Gold Probe prototype was restricted by its small-gauge needle. The second and third Injection Gold Probe prototypes had a larger-gauge needle and irrigation channel which made them faster and easier to use than separate injection catheters and thermal probes. CONCLUSIONS: The advantages of Injection Gold Probes were the ability to irrigate, inject, and coagulate without probe removal. Combination therapy did not increase treatment-related complications compared with monotherapies. PMID- 9852451 TI - Acute lower intestinal bleeding: part I: clinical presentation and diagnosis. PMID- 9852452 TI - Ileocecal tuberculosis. PMID- 9852453 TI - Ultrathin endoscopy in the assessment and treatment of upper and lower gastrointestinal tract strictures. AB - BACKGROUND: Ultrathin (1 million years, were investigated to reconstruct patterns of microbial ecological succession that have occurred since sediment burial. The relative importance of microbial transport and survival to succession was inferred from sediment ages, porewater ages, patterns of abundance (measured by direct counts, counts of culturable cells, and total phospholipid fatty acids), activities (measured by radiotracer and enzyme assays), and community composition (measured by phospholipid fatty acid patterns and Biolog substrate usage). Core samples were collected at two sites 40 km apart in the Palouse region of eastern Washington State, near the towns of Washtucna and Winona. The Washtucna site was flooded multiple times during the Pleistocene by glacial outburst floods; the Winona site elevation is above flood stage. Sediments at the Washtucna site were collected from near surface to 14.9 m depth, where the sediment age was approximately 250 ka and the porewater age was 3700 years; sample intervals at the Winona site ranged from near surface to 38 m (sediment age: approximately 1 Ma; porewater age: 1200 years). Microbial abundance and activities declined with depth at both sites; however, even the deepest, oldest sediments showed evidence of viable microorganisms. Same-age sediments had equal quantities of microorganisms, but different community types. Differences in community makeup between the two sites can be attributed to differences in groundwater recharge and paleoflooding. Estimates of the microbial community age can be constrained by porewater and sediment ages. In the shallower sediments (<9 m at Washtucna, <12 m at Winona), the microbial communities are likely similar in age to the groundwater; thus, microbial succession has been influenced by recent transport of microorganisms from the surface. In the deeper sediments, the populations may be considerably older than the porewater ages, since microbial transport is severely restricted in unsaturated sediments. This is particularly true at the Winona site, which was never flooded. PMID- 9852514 TI - Assessment of the Microbiological Potential for the Natural Attenuation of Petroleum Hydrocarbons in a Shallow Aquifer System. AB - Abstract A multidisciplinary field study investigating the fate and transport of petroleum hydrocarbons commonly associated with jet-fuel contamination is currently underway at Columbus Air Force Base (AFB), Mississippi. Sixty sediment cores from 12 boreholes were recovered from the study aquifer. The goal of this initial sampling was to characterize the potential microbial activity using 14C labeled substrates, as well as the presence, abundance, and distribution of specific hydrocarbon degrading genotypes using DNA:DNA hybridization. Enumeration of total microbial abundance using a 16S rDNA universal oligonucleotide probe was compared to traditional enumeration methods. Total culturable populations determined by spread plate analysis ranged from a low of 10(4) to more than 10(6) organisms per gram sediment. Microbial abundance estimated by DNA hybridization studies with 16S rDNA genes ranged from 10(7) to 10(8) organisms per gram sediment. Molecular analysis of aquifer samples using DNA probes targeting genes encoding the degradative enzymes alkane hydroxylase (alkB), catechol 2,3 dioxygenase (nahH), naphthalene dioxygenase (nahA), toluene dioxygenase (todC1C2), toluene monooxygenase (tomA), and xylene monooxygenase (xylA), as well as two probes measuring methanogenic microorganisms, codh (carbon monoxide dehydrogenase) and mcr (methyl coenzyme reductase), revealed that each target gene sequence was present in nearly all 60 samples. The presence of organisms demonstrating the phenotype to degrade BTEX and naphthalene was further supported using mineralization assays with 14C-labeled benzene, toluene, naphthalene, and phenanthrene. Minimal activity occurred during the first 24 hours. After a period of 5-7 days, greater than 40% of the target compounds were mineralized in aquifer sediments. PMID- 9852515 TI - A Comparison of the Use of In Vitro-Transcribed and Native rRNA for the Quantification of Microorganisms in the Environment. AB - Abstract Nearly full-length, small subunit (SSU) rRNA was transcribed in vitro from clones of SSU rDNA genes. Comparing the use of in vitro-transcribed and native rRNA indicated that, when in vitro-transcribed rRNA was used as a standard for quantitative hybridizations with oligonucleotide probes, the population was consistently underestimated. The population abundance was expressed as a percentage of specific target SSU rRNA (determined with a specific oligonucleotide probe), relative to the total SSU rRNA (measured with a universal probe). Differences in hybridization signals could be related to specific probe target locations and rRNA denaturation conditions, suggesting that higher order structure is important in quantitative membrane hybridizations. Therefore, in vitro-transcribed rRNA cannot always be used for the absolute quantification of microbial populations, but can be employed as a standard to quantify shifts in population abundance over time, and to compare community structure in various environments. PMID- 9852516 TI - Encounter with New Resources Causes Polarized Growth of the Cord-Forming Basidiomycete Phanerochaete velutina on Soil. AB - Abstract The development and physiology of cord-forming saprotrophic basidiomycetes, which form extensive and persistent mycelial networks in woodland ecosystems, can be conveniently studied on non-sterile soil in laboratory microcosms mimicking field conditions. Morphological responses of Phanerochaete velutina mycelial systems to resource encounters, and decay partitioning following encounters, varied according to whether simulated woody litter was unsterile or autoclaved and on whether encounter took place at the mycelial foraging front or behind the margin (simulating litter fall onto established systems in the field). Results show that encounter of discrete resources by P. velutina is rapidly communicated to the entire mycelial system; that resource capture takes high priority at the expense of continued system extension and decay-derived carbon reallocation; and that polarized growth toward newly encountered resources, previously considered to occur infrequently with this species, may be readily detected using image analysis techniques. Potential advantages of polarized development of P. velutina are discussed. PMID- 9852517 TI - Microbial Ecology Becomes the Official Journal of the International Society for Microbial Ecology. PMID- 9852518 TI - Factors Affecting Predation by Cyclidium sp. and Euplotes sp. on PAH-Degrading and Nondegrading Bacteria. AB - Abstract If predators select for or against contaminant-degrading bacteria, it will affect bacterial survival and has important implications for bioremediation. Protozoa are important predators of bacteria. In order to determine whether protozoa preyed differentially on bacteria with different degradation abilities, two ciliates (Euplotes sp. and Cyclidium sp.) and three strains of PAH-degrading bacteria (Vibrio spp., degrading naphthalene, anthracene, or phenanthrene) were isolated from sediment from New York/New Jersey Harbor. By manipulating growth conditions, bacterial strains with different PAH-degradation abilities and different cell properties were produced. Stepwise regression models were used to analyze how clearance rates on suspended bacteria and grazing rates on bacteria attached to particles were affected by bacterial size, hydrophobicity, C:N ratio, protein content, and PAH-degradation ability. Clearance rates ranged from 0 to 49 nl ciliate-1 h-1 for Euplotes sp. and from 0 to 1.7 nl ciliate-1 h-1 for Cyclidium sp. Clearance rates of both ciliates were positively correlated with bacterial size, hydrophobicity, and protein content, and negatively correlated with C:N ratio. PAH degradation ability had no (for Euplotes sp.) or small (for Cyclidium sp.) effects on clearance rates. The models accounted for 63-75% of the variation in clearance rates on different bacteria. Only Euplotes sp. grazed on attached bacteria, at rates from 3 to 176 bacteria ciliate-1 h-1. A regression model with only C:N ratio and protein content explained 45% of the variation in grazing rates. These models indicate that multiple properties of bacteria affect their susceptibility to predation by ciliates, but PAH-degradation ability per se has little effect. PMID- 9852519 TI - Growth Rates of Marine Ciliates on Diverse Organisms Reveal Ecological Specializations within Morphospecies. AB - Abstract The growth rate of 31 stocks of Euplotes, a cosmopolitan, marine, unicellular protist, on six food species representing two different food types, microalgae and bacteria, has been determined. The 31 stocks represented nine reproductively isolated groups (biological species?) based on breeding relationships. Three morphospecies, E. vannus, E. crassus, and E. minuta, each with both autogamous and cross-breeding breeding groups, were included. The mean number of fissions completed in 5 days of a breeding group growing on one of the six food species varied from zero to 17.06. There is a strong interaction between morphospecies and food type. The largest morphospecies, E. vannus, translates the nutritional content of algae into growth better than it translates that of bacteria, while the reverse is true for E. minuta, the smallest morphospecies. Autogamous breeding groups grow more rapidly on algae than on bacteria when compared to cross-breeding groups in the same morphospecies. Two breeding groups cannot grow on Escherichia coli. ANOVA of fissions completed in 5 days revealed significant main effects and interactions between many hierarchical levels of stocks and food species. These significant interactions indicate that genetically determined ecologically important information is present at all taxonomic levels morphospecies, breeding system, breeding group, and stock. As all these levels are biologically meaningful, measuring biodiversity in the E. vannus-crassus minuta complex solely on morphospecies will inadequately represent the ecological diversity present in the organisms and their environment. PMID- 9852520 TI - Influence of Three Contrasting Detrital Carbon Sources on Planktonic Bacterial Metabolism in a Mesotrophic Lake. AB - Abstract Lakes receive organic carbon from a diversity of sources which vary in their contribution to planktonic microbial food webs. We conducted a mesocosm study to test the effects of three different detrital carbon sources (algae, aquatic macrophytes, terrestrial leaves) on several measures of microbial metabolism in a small meso-eutrophic lake (DOC approximately 5 mg/L). Small DOC additions (DeltaC < 1 mg/L) affected bacterial numbers, growth, and pathways of carbon acquisition. Macrophyte and leaf detritus significantly increased TDP and color, but bacterial densities initially (+12 h) were unaffected. After 168 h, densities in systems amended with terrestrial detritus were 60% less than in controls, while production rates in mesocosms with macrophyte detritus were 4 fold greater. Detritus treatments resulted in greater per-cell production rates either through stable cell numbers and greater growth rates (macrophyte-C) or lower densities with stable production rates (terrestrial-C). After only 12 h, rates of leucine aminopeptidase (LAPase) activity were 2.5x greater in macrophyte C systems than in controls, but LAPase and beta-N-acetylglucosamindase activities in systems amended with terrestrial-C were only 50% of rates in controls. After 168 h, beta-xylosidase rates were significantly greater in communities with terrestrial and phytoplankton detritus. Microbial utilization of >20% of 102 carbon sources tested were affected by at least one detritus addition. Macrophyte C had positive (6% of substrates) and negative (14%) effects on substrate use; terrestrial detritus had mainly positive effects. An ordination based on carbon use profiles (+12 h) revealed a cluster of macrophyte-amended communities with greater use of psicose, lactulose, and succinamic acid; controls and algal detritus systems were more effective in metabolizing two common sugars and cellobiose. After 168 h, communities receiving terrestrial detritus were most tightly clustered, exhibiting greater use of raffinose, pyroglutamic acid, and sebacic acid. Results suggest that pelagic bacterial communities respond to changes in organic carbon source rapidly and by different routes, including shifts in per-cell production rates and variations in degradation of a variety of compounds comprising the DOC pool. PMID- 9852521 TI - Ectoenzymatic Activity and Uptake of Monomers in Marine Bacterioplankton Described by a Biphasic Kinetic Model. AB - Abstract The kinetics of bacterial hydrolytic ectoenzymatic activity and the uptake of monomeric compounds were investigated in the Northwestern Mediterranean Sea. Aminopeptidase and alpha- and beta-glucosidase activities were analyzed by using fluorogenic substrates at 15-22 concentrations ranging from 1 nM to 500 uM. Radiolabeled glucose and a mixture of amino acids were chosen as representatives of monomeric compounds, and the bacterial uptake rates (assimilation plus respiration) were determined over a wide range of substrate concentrations (from 0.2 nM to 3 uM). We found biphasic kinetics both for hydrolytic enzymes and uptake systems: high affinity enzymes at low concentrations of substrates (Km values ranged from 48 nM to 2.7 uM for ectoenzymes and from 1.4 nM to 42 nM for uptake systems), and low affinity enzymes at high concentrations of substrates (Km values ranged from 18 uM to 142 uM for ectoenzymes and from 0.1 uM to 1.3 uM for uptake systems). Transition between high and low affinity enzymes was observed at 10 uM for aminopeptidase and from 1 uM to 25 uM for glucosidases, and it was more variable and less pronounced for the uptake of glucose (40 nM-0.28 uM) and amino acids (10 nM-0.16 uM). Results showed that the potential rates of hydrolysis and uptake are tightly coupled only if the high affinity hydrolytic ectoenzymes and the low affinity uptake systems are operating simultaneously. PMID- 9852522 TI - Pelagic Bacteria-Particle Interactions and Community-Specific Growth Rates in Four Lakes along a Trophic Gradient. AB - Abstract The relationships between bacterial concentration, bacterial production, and cell-specific activity of both free and attached bacteria and environmental factors such as suspended solids, nutrients, and temperature were examined in four lakes, two in New Zealand and two in Switzerland. Estimates of cell-specific production were obtained by microautoradiographic counts of [3H]thymidine-labeled cells. Bacteria attached to particles accounted for only 1.3 to 11.6% of the total bacterial abundance, but showed overall 20-fold higher specific growth rates and were relatively more active than their free counterparts. On average, 80 to 100% of epibacteria were attached to organic particles. The abundance and production of free and attached bacteria were positively correlated; however, relationships between these fractions and some environmental variables differed. Cell-specific activities of active bacteria were not equivalent to mean cellular activities of the entire bacterial community and differed in their relationship to trophic state. [3H]Thymidine-positive bacteria were more tightly linked to chlorophyll a than were total bacteria. Our findings indicate that production by attached bacteria, fueled by phytoplankton carbon, supplies "new" free bacteria to the bacterial community. Our results support the idea that particulate organic matter acts as a source of dissolved nutrients to free bacteria. Bottom-up control of bacterial biomass, as shown by regressions of biomass vs production, appeared to be stronger in two ultraoligotrophic lakes than in two more eutrophic ones. PMID- 9852523 TI - Changes in DNA Content and Cellular Death during a Starvation-Survival Process of Escherichia coli in River Water. AB - Abstract Four nucleoid staining procedures were compared during the starvation survival process of Escherichia coli in river water. Only the method performed as a modification of the standard acridine orange direct procedure allowed us to visualize nucleoids during the 95 days of experimentation. Moreover, with this method the total number of cells and nucleoid-containing cells can be simultaneously enumerated. The decrease of the chromosomal DNA content of population and of the nucleoid-containing cells indicates that ghosts form and cellular death occurs throughout the starvation-survival process. A long time (<30 days) is needed for non-nucleoid-containing cells to appear in river water; plasmid DNA is also negatively affected by environmental stress. After 4 days of storage in river water, the need to increase the volume of lysed cells used for the plasmid band visualization as well as the decrease in the plasmid band intensity would indicate a decrease in the plasmid DNA content during the starvation-survival process. According to our results, both chromosomal and plasmid DNA content decrease during the starvation-survival process of E. coli in river water. PMID- 9852524 TI - The Impact of Oxygen Tension on Cell Density and Metabolic Diversity of Microbial Communities in Alkane Degrading Continuous-Flow Cultures. AB - Abstract The impact of the dissolved O2 tension (DOT) and the dilution rate on the metabolic diversity of an autochthonous hexadecane-degrading community in continuous-flow cultures containing hexadecane-coated intertidal sediment was determined in a set of experiments. The DOT was kept constant within each culture at values of 80% (168 umol O2L-1) or 0.4% (0.84 umol O2 L-1). The dilution rate was increased from D = 0.012 h-1 to D = 0.06 h-1. To determine the culture activity, we analyzed the hexadecane degradation rate, the protein production rate, and the oxygen consumption rate. The cell concentration of different metabolic groups was determined by colony forming units (CFU), and by most probable number (MPN). The metabolic diversity was determined by the substrate utilization spectrum in Biolog GN microtiter plates. The substrate utilization pattern of the cultures decreased considerably as D increased. This effect was more pronounced at 0.4% of DOT than at 80% of DOT. The MPN and CFU revealed that as D increased, only minor changes occurred in the community structure. The hexadecane degradation rate, the protein production rate, and the oxygen consumption rate increased parallel to D independently of the DOT. This means that the biocenosis at 0.4% of DOT was different from the biocenosis at 80% of DOT, although the metabolic activity of the cultures was unaffected by a 200 factor difference in the oxygen tension and revealed a considerable buffer capacity with respect to changes in DOT. PMID- 9852525 TI - Preoperative hearing predicts postoperative hearing. AB - Optimal hearing is one goal of otologic surgery. It is generally presumed that hearing thresholds after tympanoplasty-mastoidectomy surgery are determined by remaining anatomy. We assessed the effect of the disease process on hearing after surgery, and our data do not support this presumption. We studied the relationship between preoperative and postoperative hearing across tympanoplasties with differing anatomies in 124 patients without ossicular reconstruction with prostheses or bone. Our results indicate that poor hearing before surgery is associated with poor hearing after surgery, regardless of anatomy (type of tympanoplasty). This relationship holds for both the airbone gap and the speech-reception threshold. When considering ossicular reconstruction, the surgeon should remember that the disease process itself affects hearing in addition to the physical attributes of the ear. These results should be considered in preoperative counseling, as well as in the interpretation of the otologic literature. PMID- 9852526 TI - Epidermal growth factor and transforming growth factor-alpha in middle ear effusion. AB - Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) promote the differentiation and proliferation of epithelia as well as the proliferation and chemotaxis of fibroblasts. Additionally, EGF promotes wound healing in tissues composed largely of epithelial cells and fibroblasts. We hypothesized that EGF and TGF-alpha regulate the differentiation and proliferation of the epithelial lining and the migration and proliferation of fibroblasts in the subepithelial space of the middle ear mucosa in children with otitis media. As an initial test of this hypothesis, EGF and TGF-alpha concentrations were measured in 82 middle ear effusions of children undergoing tympanostomy tube placement. EGF was present in 45% of these effusions, and TGF alpha was present in 6%. The mean concentration +/- SEM values for EGF and TGF alpha were 19+/-7.6 and 3.7+/-7.9 pg/mL, respectively. In addition, neutrophils, macrophages, and lymphocytes in middle ear effusions stained for EGF by immunocytochemistry. We conclude that growth factors are frequently present in middle ear effusions of children with otitis media. PMID- 9852527 TI - Radiofrequency volumetric tissue reduction for treatment of turbinate hypertrophy: a pilot study. AB - OBJECTIVES: Ideal treatment for nasal obstruction caused by turbinate hypertrophy remains in question. Medical therapy is often ineffective. Surgical procedures have associated morbidity including pain, bleeding, crusting, adhesion, infection, and dryness. Radiofrequency has recently been shown to be safe and effective in volumetric tissue reduction of the tongue in the animal model and of the palate in human beings. We prospectively evaluate the safety and effectiveness of radiofrequency volumetric tissue reduction (RFVTR) for the treatment of nasal obstruction caused by inferior turbinate hypertrophy. METHODS: Twenty-two consecutive patients with nasal obstruction and associated inferior turbinate hypertrophy refractory to medical therapy were evaluated for RFVTR. The study design limited the region of treatment to the anterior third of the inferior turbinate. The procedures were performed in an ambulatory facility with patients under local anesthesia. Clinical examinations, patient questionnaires, and visual analog scales were used to assess treatment outcomes. RESULTS: No adverse effects were encountered, including bleeding, crusting, dryness, infection, adhesion, or a worsening of obstruction. Mild edema was noted in all patients but was of short duration (24 to 48 hours). Posttreatment discomfort was well controlled with acetaminophen. Eight weeks after treatment, nasal breathing improved in 21 of 22 patients, with a 58.5% reduction in severity and a 56.5% decrease in the frequency of nasal obstruction. CONCLUSION: The results of this study demonstrate that RFVTR of the hypertrophic inferior turbinate is associated with minimal adverse effects. Furthermore, this new treatment modality achieves subjective improvement in patients with symptoms of nasal obstruction. However, because of the small sample size and short follow-up, future investigations are needed to fully evaluate the use of RFVTR in the treatment of nasal obstruction caused by turbinate hypertrophy. PMID- 9852528 TI - Experimental model of cisplatin ototoxicity in chinchillas. AB - Cisplatin is an important antineoplastic agent. Its ototoxicity has been well defined, both in human and animal studies. However, animal models of systemic cisplatin administration have been complicated by multiple toxic effects. We studied cisplatin ototoxicity in an animal model involving topical application of cisplatin to the round-window membrane. Adult chinchillas were anesthetized with ketamine and pentobarbital, and auditory function was tested with the use of auditory brain-stem responses to various stimuli (clicks and 8-and 16-kHz tone bursts). Each animal was used as its own control. The middle-ear cavity was exposed through the bulla. In the experimental ear, a 25-microl solution of 0.25 mg cisplatin/1.0 ml normal saline solution was applied to the round-window membrane. In the control ear, 25 microl normal saline solution was applied to the round-window membrane. Follow-up auditory brain-stem response testing was conducted 7 days after treatment. A significant increase in threshold in the experimental ears was seen on comparison with the control ears. This finding suggests that application of cisplatin to the round-window membrane is a useful animal model in which to study cisplatin ototoxicity. PMID- 9852529 TI - Protection against aminoglycoside otic drop-induced ototoxicity by a spin trap: I. Acute effects. AB - Topical administration of aminoglycoside antibiotic drops containing neomycin and polymyxin B disrupts cochlear structure and function in rodents, possibly as a result of reactive oxygen species generation. This study investigated the ability of a spin trap, alpha-phenyl-tert-butyl-nitrone (PBN), to prevent acute aminoglycoside antibiotic drop-induced cochlear dysfunction. Guinea pigs were monitored for compound action potential thresholds and 1.0 microV root-mean square cochlear microphonic isopotential curve values, then injected intraperitoneally with PBN (60 mg/kg) or saline solution. After 10 minutes, 50 microl of PBN (100 mmol/L) or artificial perilymph was applied to the round window membrane, followed after 10 minutes with artificial perilymph or aminoglycoside antibiotic drops (50 microl). From 10 to 60 minutes after exposure, mean compound action potential thresholds progressively increased in the artificial perilymph-aminoglycoside antibiotic drop group, beginning with high frequencies and later including ever-lower frequencies. These threshold shifts in compound action potentials were significantly greater (p<0.05) than those seen in the artificial perilymph-artificial perilymph or PBN-aminoglycoside antibiotic drop groups. This finding indicates that PBN provided protection against acute aminoglycoside antibiotic drop-induced compound action potential threshold sensitivity loss. Mean cochlear microphonic shift values at 60 minutes in the artificial perilymph-aminoglycoside antibiotic drop group significantly exceeded those of the other groups only at the highest frequencies. These data suggest that acute aminoglycoside antibiotic drop-induced cochlear disruption primarily affects high frequency compound action potential function and may be partially reactive oxygen species-mediated and preventable. PMID- 9852530 TI - Auditory-nerve integrity after middle-fossa acoustic-tumor removal. AB - We sought to investigate the functional integrity of the auditory nerve in patients with postoperative hearing loss after middle cranial fossa acoustic tumor removal in a case-series descriptive study. The study setting was a tertiary referral center, a private otologic practice. The study population comprised seven patients who underwent a middle-cranial-fossa approach for unilateral acoustic-tumor resection and sustained postoperative anacusis with an anatomically intact auditory nerve. Four were men and three women; they ranged in age from 30 to 60 years; all underwent surgery between 1990 and 1994 and agreed to return to the center to participate in the study during 1995. Diagnostic electrical promontory stimulation was used to determine the functional integrity of the auditory nerve. Our main outcome measures were the presence or absence of discrete tone perception, electrical threshold, maximum acceptable level and dynamic range, gap detection and temporal difference limen during electrical promontory stimulation. Three of the seven patients demonstrated positive responses to electrical promontory testing (e.g., discrete tone perception). All three were able to perform the gap-detection and temporal difference limen tests. None of the preoperative characteristics was related to performance on promontory stimulation testing. We conclude that the middle cranial fossa approach permits anatomic--and, in some cases, functional auditory-nerve preservation. These data suggest that auditory rehabilitation in some patients who sustain anacusis after the middle cranial fossa approach to acoustic tumors, as in neurofibromatosis 2, may be provided with cochlear implantation. PMID- 9852532 TI - Fine-needle aspiration biopsy diagnosis of follicular variant of papillary thyroid cancer: therapeutic implications. AB - OBJECTIVE: Frozen-section analysis of follicular lesions is often inconclusive because capsular or vascular invasions are the hallmarks of malignancy with this technique and may not be seen in the particular field studied. Using preoperative fine-needle aspiration biopsy specimens, we attempted to identify malignant follicular lesions and compare these results against frozen-section analysis. METHODS: A retrospective study of 1000 consecutive thyroid fine-needle aspiration biopsy specimens was performed. Surgical pathologic correlation was available in 179 cases. RESULTS: Fine-needle aspiration biopsy yielded a suspicious or positive rate of 23%. Surgical pathology was available in 179 patients, of which 95% had thyroid cancer. The follicular variant of papillary cancer was identified in 26 cases, or 15% of the positive cases. Frozen-section analysis yielded false negative results in 7 of these 26 cases (27%). CONCLUSIONS: Preoperative recognition of the follicular variant of papillary cancer by fine-needle aspiration biopsy may reduce the overall incidence of false-negative frozen section findings. PMID- 9852531 TI - Inhibition of cell proliferation in head and neck squamous cell carcinoma cell lines with antisense cyclin D1. AB - Cyclin D1 and cyclin G are essential regulatory factors in the progression of the cell cycle from G0 through G1 and S phase. Aberrations in expression of these cyclins may lead to dysregulated cellular proliferation that could result in neoplasia. Amplification and overexpression of cyclin D1 have been observed in many human cancers, whereas cyclin G is a new cyclin recently described in osteosarcoma cells. This study was performed to determine whether these cyclins were amplified in head and neck squamous cell carcinoma (HNSCC) tumors. Polymerase chain reaction of DNA extracted from 22 HNSCC primary tumors and three HNSCC cell lines did not reveal amplification of cyclin D1 in any of the tumor samples. Southern blot analysis identified amplification of cyclin D1 in a single tumor. Amplification of cyclin G was not observed in any of the tumors by Southern blot hybridization with a cyclin G probe. HNSCC cell lines transfected with antisense cyclin D1 were tested for cell proliferation by the incorporation of 3H-thymidine into cells grown in serum-free media. By 72 hours of incubation, there was a greater than 30% reduction in proliferation of cells transfected with antisense cyclin D1 as compared with non-transfected control cells. The results indicate that cyclin D1 may play an important role in the growth and proliferation of HNSCC cells. PMID- 9852533 TI - Electroneurography during facial nerve expansion. AB - With the use of tissue-expansion techniques, it is possible to elongate the facial nerve without impairing its function. The rate of expansion is limited by ischemic events imposed by stretching of the nerve and by anatomic characteristics of the nerve segment. In this study, we used various electroneurography techniques to determine the first sign of facial dysfunction during expansion. The main objective of the study was to ascertain test-retest variability of electroneurography techniques. Facial nerves were expanded in 16 cats with the use of a tissue expander secured more deeply to the main trunk. We conducted electroneurography measurements with the use of surface electrodes, temporary needle electrodes, and permanently implanted electrodes. Technique dependent variations encountered with the surface electrodes made this method unreliable. Using implanted electrodes, we noted gradual worsening of the compound-action potential amplitude with increasing expansion. During acute expansion, reduction in compound-action potential amplitude was correlated with clinically observed deterioration of facial-nerve function. In this study, electroneurography with implanted electrodes was found to be the most reliable predictor of the rate of successful facial nerve expansion. PMID- 9852534 TI - Acute intracranial complications of temporal bone trauma. AB - Temporal bone trauma can be disastrous for the individual and his or her family. With the increase in violent crime throughout our society, the number of intracranial complications associated with temporal bone injury has increased significantly. Although multiple reports concerning the diagnosis and management of temporal bone trauma have been published, few studies on its intracranial complications have been addressed. In this study, we assessed the cases of 43 patients treated for temporal bone fractures between January 1992 and December 1994. The number of temporal bone fractures increased from 6 in 1992 to 21 in 1994. Twenty-one patients (49%) presented with decreased mental status. Significant alcohol and drug use was documented in 28% and 58% of patients, respectively. The most common radiologic finding was skull and intracranial injury. Thirty-six patients (84%) had at least one abnormal intracranial finding; of these patients, 14 (39%) had more than one abnormal finding on computed tomography. Cerebral midline shift, subarachnoid hemorrhage, subdural hemorrhage, and cerebral edema were the most common radiologic findings. Nineteen patients (44%) required an open neurosurgical procedure, and almost all patients with more than one abnormal radiologic finding underwent neurosurgery (86%). Seven patients required further institutional care after discharge. Four patients died (9% mortality); all the deaths were neurologically related. Mean hospital charges increased from $30,900 in 1992 to $63,000 in 1994. PMID- 9852535 TI - Plasmacytomas of the head and neck. AB - Plasmacytomas are rare tumors that often appear in the head and neck region and are characterized by a monoclonal proliferation of plasma cells. On both clinical presentation and pathologic examination these tumors may be confused with more common tumors of the head and neck. The purpose of this article is to review our experience with these rare neoplasms, with emphasis on clinical, pathologic, and therapeutic features. On retrospective chart review, we identified 20 patients with the diagnosis of plasmacytoma of the head and neck region at the Cleveland Clinic Foundation between 1976 and 1993. Records were reviewed with regard to initial symptoms, location of the neoplasm, diagnostic evaluation, treatment modalities, and survival. Of the 20 cases we identified, the tumor arose in the sinonasal/nasopharyngeal region in 11 (55%). Two cases (10%) represented medullary plasmacytomas, arising in the clavicle and presenting as supraclavicular masses. The mean follow-up was 60.2 months (range 6 to 131 months). In 15 of the 20 cases, immunohistochemistry staining for immunoglobulin light chain production was conducted. One of the two cases (50%) classified as medullary plasmacytoma demonstrated conversion to multiple myeloma, whereas only 2 of 18 cases of extramedullary plasmacytoma (11%) converted to multiple myeloma. The primary modality of treatment was radiation therapy with typical doses of 4500 to 6000 cGy. Kaplan-Meier survival estimates demonstrated 95% survival at 1 year, 82% survival at 5 years, and 10-year estimated survival of 72%. Plasmacytomas of the head and neck region are rare and on initial evaluation must be distinguished from multiple myeloma. The diagnostic evaluation includes appropriate radiologic and pathologic studies including immunohistochemistry. Despite the typical presentation as a locally destructive tumor, plasmacytomas are highly radiosensitive, and 70% to 80% survival may be obtained with the use of radiotherapy. Patients with plasmacytomas require long-term follow-up to detect conversion to multiple myeloma. PMID- 9852536 TI - Metachronous cancer: prognostic factors including prior irradiation. AB - In this article we evaluate two factors that may be responsible for the reported increased mortality rate in metachronous cancers: prior radiation therapy and stage at presentation. A select group of 358 patients was split into three groups: no prior cancer (group 1), prior cancer treated with radiation therapy (group 2), and prior cancer treated with surgery alone (group 3). We compared survival among the three groups according to stage (T1 or T2 vs. T3 or T4) using the Lifetest procedure. Survival in patients with advanced (T3 or T4) cancers was uniformly poor, and survival in patients with low-staged (T1 or T2) cancers was disproportionately poor only for patients in group 2. Metachronous cancers are not necessarily more lethal, except when the cancer arises within prior irradiated tissue. Initial treatment decisions for patients with primary cancers must always provide for the contingency of a metachronous cancer, and the judicious use of radiation therapy is essential. PMID- 9852537 TI - Computed tomography in the evaluation of pediatric neck infections. AB - In children, infections involving both the superficial and deep neck spaces are common. Children so affected typically present with fever, neck mass, neck stiffness, and, occasionally, airway compromise. Radiologic modalities used in the evaluation of neck infections include plain lateral neck radiography, ultrasound, computed tomography, and magnetic resonance imaging. All these modalities have proved useful in the treatment of such infections, specifically the decision to perform incision and drainage. The charts of 66 patients-33 with superficial and 33 with deep neck infections-were analyzed with respect to symptoms, signs, computed tomography findings, and need for surgical intervention. Computed tomography was not particularly helpful in superficial neck infections with regard to the decision to perform surgical drainage; however, it did localize and demonstrate the extent of infection. In deep neck infections we found a 92% correlation between computed tomographic evidence of an abscess and surgical confirmation of one. Contrast-enhanced computed tomography remains an excellent tool in the treatment of neck infections in children. PMID- 9852538 TI - Influence of computed tomography on pretherapeutic tumor staging in head and neck cancer patients. AB - Refinements in radiographic techniques have resulted in increased use of radiographic studies in the evaluation of patients with head and neck cancer over the past 20 years. To assess the impact of such studies, we compared tumor clinical stages based solely on physical-examination findings with those obtained with the addition of CT findings. This study was accomplished through case review of 81 head and neck cancer patients who underwent CT after preliminary TNM-stage assignment as determined on the basis of physical examination alone. In this cohort, 44 patients (54%) had a change in assigned clinical stage. We reviewed individual anatomic sites to determine where CT was found to be most useful in modifying tumor stage. Changes in tumor and nodal stage were found across all major sites of the head and neck. Tumors of the hypopharynx were the most likely to change stage (90%) on the basis of CT findings, whereas tumors of the glottic larynx were least likely to undergo a change in stage (16%). The therapeutic implications of these findings are discussed in the context of the published literature. PMID- 9852539 TI - Biomechanics of combined arytenoid adduction and medialization laryngoplasty in an ex vivo canine model. AB - Arytenoid adduction (AA) and medialization laryngoplasty (ML) are being performed concurrently in patients with unilateral vocal fold paralysis with a large posterior glottal gap. The biomechanical effects of this combined procedure on the larynx have not been studied. An excised canine larynx model was used to study the effects of AA, ML, and combined AA and ML (AA-ML) on vocal fold configuration (length and degree of medialization) and tension. AA-ML lengthens the affected vocal fold relative to the opposite vocal fold, although both were slightly shortened compared with the control state (nonsignificant trends). AA-ML medializes the entire length of the vocal fold more effectively than AA or ML alone. Midmembranous vocal fold tension did not increase with AA-ML. With AA-ML, the vocal process resisted significantly greater lateralizing forces than with the control state or ML. We conclude that in an excised canine model, AA-ML combines the biomechanical properties of AA and ML. The larynx appears to be divided into 2 biomechanical subunits: membranous vocal fold (anterior) and arytenoid cartilage (posterior). When surgical rehabilitation of both laryngeal subunits is required, AA-ML appears to be a better choice than AA or ML alone. PMID- 9852540 TI - Efficacy of laser-assisted uvulopalatoplasty in obstructive sleep apnea. AB - Laser-assisted uvulopalatoplasty (LAUP) enlarges the oropharyngeal airway by reshaping the uvula, soft palate, and tonsillar pillars. In contrast to a uvulopalatopharyngoplasty performed in the operating room, LAUP is staged over several office visits, is less costly, is bloodless, and is not associated with velopharyngeal insufficiency or stenosis. Previous studies have documented the efficacy of LAUP in the treatment of snoring. We explore the efficacy of LAUP in the treatment of obstructive sleep apnea. Forty-eight patients comprised the study group. Each patient snored, exhibited >50% palatal collapse on Muller's maneuver, and had nocturnal polysomnography showing a respiratory disturbance index (RDI) >10. Each patient underwent LAUP until he or she reported that snoring had ceased. Of the 29 patients who underwent post-LAUP nocturnal polysomnography, 7 had RDIs <10, oxygen saturation >86%, and no cardiac arrhythmias. LAUP reduced RDI to <10 in patients with pre-LAUP apnea indexes <3 (P = 0.05) or pre-LAUP RDIs <30 (P = 0.01). The following variables did not correlate with the success of LAUP in treating obstructive sleep apnea: age, sex, pre-LAUP weight, pre-LAUP body mass index, perioperative weight change, perioperative body mass index change, pre-LAUP snoring, and post-LAUP snoring. We conclude that LAUP is an effective means of treating patients with RDIs <30. PMID- 9852541 TI - Immunotherapy for allergic fungal sinusitis: three years' experience. AB - Since August 1994, we have treated patients with histologically proven allergic fungal sinusitis with surgery followed by immunotherapy, employing fungal and nonfungal antigens to which hypersensitivity has been demonstrated. Our results continue to be encouraging. Not only have we encountered no indication that fungal immunotherapy has worsened these patients' condition or caused a recurrence of disease, we have confirmed dramatic improvement in these patients compared with the generally accepted course of this disease. Of 11 patients who have received immunotherapy for 1 to 3 years (mean 28 months), none has required regular or frequent treatment with a single brief course of systemic steroids, and only three are receiving topical nasal steroids. No repeat surgeries for recurrent allergic fungal sinusitis have been required in the treatment group. This combination of surgery and immunotherapy has continued to prove beneficial, and we urge others to consider this approach to therapy. PMID- 9852542 TI - Diffuse neurofibroma of the orbit associated with temporal meningocele and neurofibromatosis-1. PMID- 9852543 TI - Basaloid squamous cell carcinoma and fine-needle aspiration: a potential diagnostic pitfall. PMID- 9852544 TI - Meningiomas presenting in the temporal bone: the pathways of spread from an intracranial site of origin. PMID- 9852545 TI - Juvenile aggressive ossifying fibroma presenting as an ethmoid sinus mucocele. PMID- 9852546 TI - Facial nerve dysfunction associated with cystic lesions of the mastoid. PMID- 9852547 TI - Oral lymphoma in human immunodeficiency virus infection: a report of six cases and review of the literature. PMID- 9852549 TI - Intracochlear schwannoma. PMID- 9852548 TI - Ammonia capsule ingestion causing upper aerodigestive tract injury. PMID- 9852550 TI - Subcochlear petrous cholesterol granuloma involving the infratemporal fossa. PMID- 9852551 TI - Air in the vestibule: computed tomography scan finding in traumatic perilymph fistula. PMID- 9852552 TI - Simple, new device for needle aspiration biopsy. AB - Vertigo and dizziness are not common in childhood, but are probably present more often than was formerly thought. These symptoms caused mainly by o700is media and middle ear effusion, two of the most common diseases in children, have been neglected for a long time, both in the literature and in practice, until recently. The purpose of this study was to determine objectively the incidence of balance-related symptoms in children with long-lasting middle ear effusion and to discover whether these symptoms resolve after the insertion of ventilation tubes. One hundred thirty-six children, ages 4 to 9 years, were given electronystagmographic tests and the Bruininks-Oseretsky tests for motor proficiency before and after tube ventilation of the middle ear. The results were compared with those in 74 healthy children with no history of middle ear diseases. Pathologic findings were found in 58% of the children with chronic middle ear effusion, as compared with only 4% of the control group. The symptoms and signs of balance disturbances resolved in 96% of the children after ventilation tube insertion. The results of this study indicate that balance related symptoms often encountered in young children may result from chronic middle ear effusion and that these symptoms will resolve after evacuation of the effusion and ventilation of the middle ear. PMID- 9852553 TI - Effects of middle ear effusion on the vestibular system in children. AB - Vertigo and dizziness are not common in childhood, but are probably present more often than was formerly thought. These symptoms caused mainly by otitis media and middle ear effusion, two of the most common diseases in children, have been neglected for a long time, both in the literature and in practice, until recently. The purpose of this study was to determine objectively the incidence of balance-related symptoms in children with long-lasting middle ear effusion and to discover whether these symptoms resolve after the insertion of ventilation tubes. One hundred thirty-six children, ages 4 to 9 years, were given electronystagmographic tests and the Bruininks-Oseretsky tests for motor proficiency before and after tube ventilation of the middle ear. The results were compared with those in 74 healthy children with no history of middle ear diseases. Pathologic findings were found in 58% of the children with chronic middle ear effusion, as compared with only 4% of the control group. The symptoms and signs of balance disturbances resolved in 96% of the children after ventilation tube insertion. The results of this study indicate that balance related symptoms often encountered in young children may result from chronic middle ear effusion and that these symptoms will resolve after evacuation of the effusion and ventilation of the middle ear. PMID- 9852554 TI - Sucralfate for posttonsillectomy analgesia. AB - Pain is one of the most troublesome complications of tonsillectomy. The pain appears as throat pain, otalgia, or both, and continues until mucosal recovery on the tonsillar fossae is complete. Some surgical and hemostasis techniques may increase pain. Analgesics, antibiotics, steroids, and local and topical anesthetics are used to relieve posttonsillectomy pain, but none has the desired effectiveness. The pain reliever must not increase bleeding and must have minimal side effects. Sucralfate, a basic amino salt of sucrose octasulfate, binds to the matrix protein of a peptic ulcer and produces a protective barrier. Tonsillectomy leaves two large ulcerous wounds, and sucralfate may bind those wounds as it does peptic ulcers. In this controlled study, the efficacy of sucralfate on posttonsillectomy throat pain, otalgia, analgesic requirement, degree of strength, bleeding, body temperature, and mucosal recovery is investigated in 80 patients. Sucralfate is found to significantly reduce throat pain and analgesic requirement after surgery. PMID- 9852555 TI - Retropharyngeal abscess associated with vertebral osteomyelitis and spinal epidural abscess. PMID- 9852556 TI - Tonsil cancer and Sweet's syndrome. PMID- 9852557 TI - Intramucosal spread of malignant melanoma of the oral cavity. PMID- 9852558 TI - Unilateral laryngeal dysgenesis. PMID- 9852559 TI - Elisa comparison of three whole-cell antigens of Borrelia burgdorferi sensu lato in serological study of dogs from area of Kosice, eastern Slovakia. AB - Three antigens of Borrelia burgdorferi sensu lato (B. burgdorferi sensu stricto- Slovak strain Ir 105, B. garini - Slovak strain K 24 and B. burgdorferi sensu stricto--American strain B 31) were compared by ELISA on a group of dogs from urban agglomeration of Kosice, eastern Slovakia. Of 256 serum samples from dogs examined for the presence of anti-Borrelia IgG antibodies, 128 (50%) were positive with Ir 105 antigen, 107 (41.7%) with K 24 and 74 (28.9%) with B 31. The seroprevalence between strains B 31 and K 24 and B 31 and Ir 105 differed statistically significantly (test chi2, p < 0.05), however, the difference between strains K 24 and Ir 105 was insignificant. A significantly higher seroprevalence of all the strains examined was detected in hunting dogs (test chi2, p < 0.05) when compared with service and pet dogs. The seroprevalence correlated with the frequency of outing the dogs in woody areas with the occurrence of borreliae in ticks (R = 0.5 or 0.7) as well as with the frequency of finding engorged ticks (R = 0.5). An epizootiological anamnesis showed a fair specificity in all the strains examined. The Slovak strains showed the higher consistency of positive and negative findings (70%) but statistically lower specificity than the American strain. PMID- 9852561 TI - Synaptic physiology and ultrastructure in comatose mutants define an in vivo role for NSF in neurotransmitter release. AB - N-Ethylmaleimide-sensitive fusion protein (NSF) is a cytosolic protein thought to play a key role in vesicular transport in all eukaryotic cells. Although NSF was proposed to function in the trafficking of synaptic vesicles responsible for neurotransmitter release, only recently have in vivo experiments begun to reveal a specific function for NSF in this process. Our previous work showed that mutations in a Drosophila NSF gene, dNSF1, are responsible for the temperature sensitive paralytic phenotype in comatose (comt) mutants. In this study, we perform electrophysiological and ultrastructural analyses in three different comt alleles to investigate the function of dNSF1 at native synapses in vivo. Electrophysiological analysis of postsynaptic potentials and currents at adult neuromuscular synapses revealed that in the absence of repetitive stimulation, comt synapses exhibit wild-type neurotransmitter release at restrictive (paralytic) temperatures. In contrast, repetitive stimulation at restrictive temperatures revealed a progressive, activity-dependent reduction in neurotransmitter release in comt but not in wild type. These results indicate that dNSF1 does not participate directly in the fusion of vesicles with the target membrane but rather functions in maintaining the pool of readily releasable vesicles competent for fast calcium-triggered fusion. To define dNSF1 function further, we used transmission electron microscopy to examine the distribution of vesicles within synaptic terminals, and observed a marked accumulation of docked vesicles at restrictive temperatures in comt. Together, the results reported here define a role for dNSF1 in the priming of docked synaptic vesicles for calcium-triggered fusion. PMID- 9852560 TI - Enhancement of neurotransmitter release induced by brain-derived neurotrophic factor in cultured hippocampal neurons. AB - Brain-derived neurotrophic factor (BDNF), like other neurotrophins, has long-term effects on neuronal survival and differentiation; furthermore, recent work has shown that BDNF also can induce rapid changes in synaptic efficacy. We have investigated the mechanism(s) of these synaptic effects on cultured embryonic hippocampal neurons. In the presence of the GABAA receptor antagonist, picrotoxin, the application of BDNF (100 ng/ml) for 1-5 min increased the amplitude of evoked synaptic currents by 48 +/- 9% in 10 of 15 pairs of neurons and increased the frequency of EPSC bursts to 205 +/- 20% of the control levels. There was no detectable effect of BDNF on various measures of electrical excitability, including the resting membrane potential, input resistance, action potential threshold, and action potential amplitude. In addition, BDNF did not change the postsynaptic currents induced by the exogenous application of glutamate. BDNF did increase the frequency of miniature EPSCs (mEPSCs) (268.0 +/- 46.8% of control frequency), however, without affecting the mEPSC amplitude. The effect of BDNF on mEPSC frequency was blocked by the tyrosine kinase inhibitor K252a and also by the removal of extracellular calcium ([Ca2+]o). Fura-2 recordings showed that BDNF elicited an increase in intracellular calcium concentration ([Ca2+]c). This effect was dependent on [Ca2+]o; it was blocked by K252a and by thapsigargin, but not by caffeine. The results demonstrate that BDNF enhances glutamatergic synaptic transmission at a presynaptic locus and that this effect is accompanied by a rise in [Ca2+]c that requires the release of Ca2+ from IP3-gated stores. PMID- 9852562 TI - NSF function in neurotransmitter release involves rearrangement of the SNARE complex downstream of synaptic vesicle docking. AB - The SNARE hypothesis has been proposed to explain both constitutive and regulated vesicular transport in eukaryotic cells, including release of neurotransmitter at synapses. According to this model, a vesicle targeting/docking complex consisting primarily of vesicle- and target-membrane proteins, known as SNAREs, serves as a receptor for the cytosolic N-ethylmaleimide-sensitive fusion protein (NSF). NSF dependent hydrolysis of ATP disassembles the SNARE complex in a step postulated to initiate membrane fusion. While features of this model remain tenable, recent studies have challenged fundamental aspects of the SNARE hypothesis, indicating that further analysis of these components is needed to fully understand their roles in neurotransmitter release. We have addressed this issue by using the temperature-sensitive Drosophila NSF mutant comatose (comt) to study the function of NSF in neurotransmitter release in vivo. Synaptic electrophysiology and ultrastructure in comt mutants have recently defined a role for NSF after docking in the priming of synaptic vesicles for fast calcium-triggered fusion. Here we report that an SDS-resistant neural SNARE complex, composed of the SNARE polypeptides syntaxin, n-synaptobrevin, and SNAP-25, accumulates in comt mutants at restrictive temperature. Subcellular fractionation experiments indicate that these SNARE complexes are distributed predominantly in fractions containing plasma membrane and docked synaptic vesicles. Together with the electrophysiological and ultrastructural analyses of comt mutants, these results indicate that NSF functions to disassemble or otherwise rearrange a SNARE complex after vesicle docking and that this rearrangement is required to maintain the readily releasable pool of synaptic vesicles. PMID- 9852563 TI - Cocaine acts as an apparent competitive inhibitor at the outward-facing conformation of the human norepinephrine transporter: kinetic analysis of inward and outward transport. AB - The inhibition by cocaine of inward and outward transport of dopamine (DA) at the cloned human norepinephrine transporter (hNET) and the relationship of the inhibitory patterns of cocaine to the conformational requirements of the transporter were investigated. This was done using rotating disk electrode voltammetry in transfected cells. The uphill uptake of external DA, the lack of inhibition by internal substrates on DA uptake, and the accelerated exchange of internal DA by external m-tyramine support a carrier model in which the hNET alternates between outward-facing and inward-facing conformations. Cocaine exhibited competitive inhibition of DA uptake, which was insensitive to intracellular substrates. In contrast, the inhibition by cocaine of the m tyramine-induced DA efflux appeared noncompetitive relative to intracellular DA, but competitive relative to extracellular m-tyramine. Simultaneous measurement of m-tyramine uptake and accompanying DA efflux at various concentrations of intracellular DA showed that cocaine did not alter the ratio of DA efflux to m tyramine uptake. Moreover, cocaine displayed similar potency for inhibiting DA uptake and efflux. Additionally, the inhibition profile of cocaine was unrelated to the addition time of cocaine, simultaneously with or earlier than a substrate. All of the findings are consonant with a competitive interaction between cocaine and substrates at the outward-facing conformation of the hNET. This action directly prevents the inward transport of external substrates, thereby inhibiting the outward transport of internal substrates by reducing the availability of the inward-facing conformation. Consequently, the experimental inhibition pattern of cocaine depends on the conformation of the hNET to which the transported substrate is exposed. PMID- 9852565 TI - Mitochondrial control of acute glutamate excitotoxicity in cultured cerebellar granule cells. AB - Mitochondria within cultured rat cerebellar granule cells have a complex influence on cytoplasmic free Ca2+ ([Ca2+]c) responses to glutamate. A decreased initial [Ca2+]c elevation in cells whose mitochondria are depolarized by inhibition of the ATP synthase and respiratory chain (conditions which avoid ATP depletion) was attributed to enhanced Ca2+ extrusion from the cell rather than inhibited Ca2+ entry via the NMDA receptor. Even in the presence of elevated extracellular Ca2+, when [Ca2+]c responses were restored to control values, such cells showed resistance to acute excitotoxicity, defined as a delayed cytoplasmic Ca2+ deregulation (DCD) during glutamate exposure. DCD was a function of the duration of mitochondrial polarization in the presence of glutamate rather than the total period of glutamate exposure. Once initiated, DCD could not be reversed by NMDA receptor inhibition. In the absence of ATP synthase inhibition, respiratory chain inhibitors produced an immediate Ca2+ deregulation (ICD), ascribed to an ATP deficit. In contrast to DCD, ICD could be reversed by subsequent ATP synthase inhibition with or without additional NMDA receptor blockade. DCD could not be ascribed to the failure of an ATP yielding metabolic pathway. It is concluded that mitochondria can control Ca2+ extrusion from glutamate-exposed granule cells by the plasma membrane in three ways: by competing with efflux pathways for Ca2+, by restricting ATP supply, and by inducing a delayed failure of Ca2+ extrusion. Inhibitors of the mitochondrial permeability transition only marginally delayed the onset of DCD. PMID- 9852564 TI - Enhanced opioid efficacy in opioid dependence is caused by an altered signal transduction pathway. AB - Chronic morphine administration induces adaptations in neurons resulting in opioid tolerance and dependence. Functional studies have implicated a role for the periaqueductal gray area (PAG) in the expression of many signs of opioid withdrawal, but the cellular mechanisms are not fully understood. This study describes an increased efficacy, rather than tolerance, of opioid agonists at mu receptors on GABAergic (but not glutamatergic) nerve terminals in PAG after chronic morphine treatment. Opioid withdrawal enhanced the amplitudes of electrically evoked inhibitory synaptic currents mediated by GABAA receptors and increased the frequency of spontaneous miniature GABAergic synaptic currents. These effects were not blocked by 4-aminopyridine or dendrotoxin, although both Kv channel blockers abolish acute opioid presynaptic inhibition of GABA release in PAG. Instead, the withdrawal-induced increases were blocked by protein kinase A inhibitors and occluded by metabolically stable cAMP analogs, which do not prevent acute opioid actions. These findings indicate that opioid dependence induces efficacious coupling of mu-receptors to presynaptic inhibition in GABAergic nerve terminals via adenylyl cyclase- and protein kinase A-dependent processes in PAG. The potential role of these adaptations in expression of withdrawal behavior was supported by inhibition of enhanced GABAergic synaptic transmission by the alpha2 adrenoceptor agonist clonidine. These findings provide a cellular mechanism that is consistent with other studies demonstrating attenuated opioid withdrawal behavior after injections of protein kinase A inhibitors into PAG and suggest a general mechanism whereby opioid withdrawal may enhance synaptic neurotransmission. PMID- 9852566 TI - Thiol oxidation and loss of mitochondrial complex I precede excitatory amino acid mediated neurodegeneration. AB - Human ingestion of "chickling peas" from the plant Lathyrus sativus, which contains an excitatory amino acid, L-BOAA (L-beta-N-oxalylamino-L-alanine), leads to a progressive corticospinal neurodegenerative disorder, neurolathyrism. Exposure to L-BOAA, but not its optical enantiomer D-BOAA, causes mitochondrial dysfunction as evidenced by loss of complex I activity in vitro in male mouse brain slices and in vivo in selected regions of mouse CNS (lumbosacral cord and motor cortex). Loss of complex I activity in lumbosacral cord after L-BOAA administration to mice was accompanied by concurrent loss of glutathione. The inhibited complex I activity in mitochondria isolated from lumbosacral cord of animals treated with L-BOAA rebounded after incubation with the thiol-reducing agent dithiothreitol, indicating that oxidation of protein thiols to disulfides was responsible for enzyme inhibition. The inhibition of complex I could be abolished by pretreatment with antioxidant thiols such as glutathione ester and alpha-lipoic acid. Chronic treatment of male mice, but not female mice, with L BOAA resulted in loss of complex I activity and vacuolation and dendritic swelling of neurons in the motor cortex and lumbar cord, paralleling the regionality of the aforementioned biochemical effects on CNS mitochondria. These results support the view that thiol oxidation and concomitant mitochondrial dysfunction (also implicated in other neurodegenerative disorders), occurring downstream of glutamate receptor activation by L-BOAA, are primary events leading to neurodegeneration. Maintenance of protein thiol homeostasis by thiol delivery agents could potentially offer protection against excitotoxic insults such as those seen with L-BOAA. PMID- 9852567 TI - A dopamine/D1 receptor/protein kinase A/dopamine- and cAMP-regulated phosphoprotein (Mr 32 kDa)/protein phosphatase-1 pathway regulates dephosphorylation of the NMDA receptor. AB - We have investigated the mechanism by which activation of dopamine (DA) receptors regulates the glutamate sensitivity of medium spiny neurons of the nucleus accumbens. Our results demonstrate that DA regulates the phosphorylation state of the NR1 subunit of NMDA-type glutamate receptors. The effect of DA was mimicked by SKF82526, a D1-type DA receptor agonist, and by forskolin, an activator of cAMP-dependent protein kinase (PKA), and was blocked by H-89, a PKA inhibitor. These data indicate that DA increases NR1 phosphorylation through a PKA-dependent pathway. DA-induced phosphorylation of NR1 was blocked in mice bearing a targeted deletion of the gene for dopamine- and cAMP-regulated phosphoprotein of Mr 32 kDa (DARPP-32), a phosphoprotein that is a potent and selective inhibitor of protein phosphatase-1, indicating that the effect of PKA is mediated, in part, by regulation of the DARPP-32/protein phosphatase-1 cascade. In support of this interpretation, NR1 phosphorylation was increased by calyculin A, a protein phosphatase-1/2A inhibitor. A model is proposed in which the ability of DA to regulate NMDA receptor sensitivity is attributable to a synergistic action involving increased phosphorylation and decreased dephosphorylation of the NR1 subunit of the NMDA receptor. PMID- 9852568 TI - Protein kinase A activity may kinetically upregulate the striatal transporter for dopamine. AB - The neuronal dopamine transporter (DAT) plays a key role in terminating dopaminergic chemical neurotransmission; thus, the study of the regulation of DAT activity is important in defining parameters relevant to the control of dopaminergic neurotransmission. Interpretation of the results from previous work of this laboratory suggests that occupation of presynaptic autoreceptors increases DAT activity. Second messenger signaling related to kinetic upregulation of DAT has not been examined previously. However, others have shown that protein kinase C activity may downregulate DAT activity, whereas protein kinase A has shown variable results. Herein it is shown that protein kinase A activity mediates the kinetic upregulation of DAT. Quinpirole increased DAT activity that was blocked by sulpiride and the protein kinase A selective inhibitor H-89. Brief incubations with forskolin and 8-bromo-cAMP (8-Br-cAMP) were found to stimulate striatal DAT activity by increasing the Vmax of transport without affecting the Km. Exposures >15 min had no effect. The 8-Br-cAMP stimulated increases in DAT activity were blocked by pre-exposure to H-89. Thus, second messenger signaling via the cAMP cascade may mediate kinetic upregulation of DAT. Kinetic analyses of the results suggest that either insertion of DAT into the membrane or activation of pre-existing DAT within the membrane mediates the regulation. PMID- 9852569 TI - Use-dependent decline of paired-pulse facilitation at Aplysia sensory neuron synapses suggests a distinct vesicle pool or release mechanism. AB - We have characterized paired-pulse facilitation at Aplysia sensory neuron-to motoneuron synapses. This simple form of very short-term synaptic plasticity displayed an unusual feature: it decreased dramatically with repeated testing. Synaptic depression at these synapses and this use-dependent decrease in paired pulse facilitation occurred independently of each other. Paired-pulse facilitation was inversely correlated with the size of the initial synaptic connection and was absent at stronger synapses. The use-dependent decrease in paired-pulse facilitation occurred at the same rate at large synapses as at small synapses, although the initial paired-pulse facilitation at large synapses was substantially smaller. Rates of synaptic depression were also independent of initial synaptic strength. Paired-pulse facilitation was blocked by presynaptic EGTA injection, but not by postsynaptic EGTA or BAPTA injection. These results indicate that presynaptic Ca2+ influx plays a critical role in paired-pulse facilitation. However, the persistence of the decrease in paired-pulse facilitation for longer than 15 min suggests that Ca2+ from the first paired action potential produces facilitation via a modulatory mechanism rather than by summating with Ca2+ influx during the second paired action potential in activating the Ca2+ binding sites that initiate exocytosis. This modulatory mechanism may not involve protein phosphorylation because paired-pulse facilitation was unaffected by the protein kinase inhibitors H7 and KN-62. These findings further suggest that release by the second paired action potential occurs at sites distinct from those that mediate release by the first action potential. PMID- 9852570 TI - Gating of the L-type Ca channel in human skeletal myotubes: an activation defect caused by the hypokalemic periodic paralysis mutation R528H. AB - The skeletal muscle L-type Ca channel serves a dual role as a calcium-conducting pore and as the voltage sensor coupling t-tubule depolarization to calcium release from the sarcoplasmic reticulum. Mutations in this channel cause hypokalemic periodic paralysis (HypoPP), a human autosomal dominant disorder characterized by episodic failure of muscle excitability that occurs in association with a decrease in serum potassium. The voltage-dependent gating of L type Ca channels was characterized by recording whole-cell Ca currents in myotubes cultured from three normal individuals and from a patient carrying the HypoPP mutation R528H. We found two effects of the R528H mutation on the L-type Ca current in HypoPP myotubes: (1) a mild reduction in current density and (2) a significant slowing of the rate of activation. We also measured the voltage dependence of steady-state L-type Ca current inactivation and characterized, for the first time in a mammalian preparation, the kinetics of both entry into and recovery from inactivation over a wide range of voltages. The R528H mutation had no effect on the kinetics or voltage dependence of inactivation. PMID- 9852571 TI - Mammalian nicotinic receptors with alpha7 subunits that slowly desensitize and rapidly recover from alpha-bungarotoxin blockade. AB - One of the most abundant nicotinic receptors in the nervous system is a species that contains the alpha7 gene product, rapidly desensitizes, and binds alpha bungarotoxin with great affinity. The receptor has a high relative permeability to calcium and performs a variety of functions including presynaptic modulation of transmitter release and postsynaptic generation of synaptic currents. Fast excitatory transmission in mammalian intracardiac ganglia is mediated primarily by nicotinic receptors, and although intracardiac ganglion neurons express the alpha7 gene, no toxin-sensitive response has been detected previously in them. We report here that whole-cell patch-clamp recordings from freshly dissociated intracardiac ganglion neurons reveal a nicotinic response that desensitizes slowly and is blocked by alpha-bungarotoxin in a rapidly reversible manner. The only rat gene previously thought capable of forming such receptors was alpha9, but no evidence suggests that the alpha9 gene is expressed in neurons. We find that reverse transcription (RT)-PCR detects alpha7 but not alpha9 mRNA in the ganglia. In addition, the pharmacology of the nicotinic response is typical of alpha7-containing receptors but differs in several respects from that expected for alpha9. Binding experiments with immunotethered receptors identifies a ganglionic species that contains the alpha7 gene product. Moreover, intracellular perfusion of the cells with an anti-alpha7 monoclonal antibody specifically reduces the amplitude of the toxin-sensitive response. The results indicate that alpha7-containing receptors are responsible for the slowly desensitizing, toxin reversible response and suggest that the receptors are modified in cell-specific ways to influence their functional properties. PMID- 9852572 TI - Modulation of TTX-R INa by PKC and PKA and their role in PGE2-induced sensitization of rat sensory neurons in vitro. AB - A tetrodotoxin-resistant voltage-gated Na+ current (TTX-R INa) appears to be the current primarily responsible for action potential generation in the cell body and terminals of nociceptive afferents. Although other voltage-gated Na+ currents are modulated by the activation of protein kinase C (PKC), protein kinase A (PKA), or both, the second messenger pathways involved in the modulation of TTX-R INa are still being defined. We have examined the modulation of TTX-R INa in isolated sensory neurons with whole-cell voltage-clamp recording. Activation of either PKC or PKA increased TTX-R INa. PKA activation also produced a leftward shift in the conductance-voltage relationship of TTX-R INa and an increase in the rates of current activation, deactivation, and inactivation. Inhibitors of PKC decreased TTX-R INa, whereas inhibitors of PKA had no effect on the current. Investigating the interaction between PKC and PKA revealed that although inhibitors of PKA had little effect on PKC-induced modulation of TTX-R INa, inhibitors of PKC significantly attenuated PKA-induced modulation of the current. Finally, although PGE2-induced modulation of TTX-R INa was more similar to PKA induced modulation of the current than to PKC-induced modulation, PGE2-induced effects were inhibited by inhibitors of both PKC and PKA. Thus, although TTX-R INa is a common target for cellular processes involving the activation of either PKA or PKC, PKC activity is necessary to enable subsequent PKA-mediated modulation of TTX-R INa. PMID- 9852573 TI - Nerve growth factor-dependent activation of NF-kappaB contributes to survival of sympathetic neurons. AB - Neurotrophins activate multiple signaling pathways in neurons. However, the precise roles of these signaling molecules in cell survival are not well understood. In this report, we show that nerve growth factor (NGF) activates the transcription factors NF-kappaB and AP-1 in cultured sympathetic neurons. Activated NF-kappaB complexes were shown to consist of heterodimers of p50 and Rel proteins (RelA, as well as c-Rel), and NF-kappaB activation was found to occur independently of de novo protein synthesis but in a manner that required the action of the proteasome complex. Treatment with the NF-kappaB inhibitory peptide SN50 in the continuous presence of NGF resulted in dose-dependent induction of cell death. Under the conditions used, SN50 was shown to selectively inhibit NF-kappaB activation but not the activation of other cellular transcription factors such as AP-1 and cAMP response element-binding protein. Cells treated with SN50 exhibited morphological and biochemical hallmarks of apoptosis, and the kinetics of cell killing were accelerated relative to death induced by NGF withdrawal. Finally, experiments were conducted to test directly whether NF-kappaB could act as a survival factor for NGF-deprived neurons. Microinjection of cells with an expression plasmid encoding NF-kappaB (c-Rel) resulted in enhanced neuronal survival after withdrawal of NGF, whereas cells that were transfected with a vector encoding a mutated derivative of c-Rel lacking the transactivation domain underwent cell death to the same extent as control cells. Together, these findings suggest that the activation of NF kappaB/Rel transcription factors may contribute to the survival of NGF-dependent sympathetic neurons. PMID- 9852574 TI - Effects of transforming growth factor-beta (isoforms 1-3) on amyloid-beta deposition, inflammation, and cell targeting in organotypic hippocampal slice cultures. AB - The transforming growth factor-beta (TGF-beta) family consists of three isoforms and is part of a larger family of cytokines regulating differentiation, development, and tissue repair. Previous work from our laboratory has shown that TGF-beta1 can increase amyloid-beta protein (Abeta) immunoreactive (Abetair) plaque-like deposits in rat brain. The aim of the current study was to evaluate all three isoforms of TGF-beta for their ability to affect the deposition and neurotoxicity of Abeta in an organotypic, hippocampal slice culture model of Abeta deposition. Slice cultures were treated with Abeta either with or without one of the TGF-beta isoforms. All three isoforms can increase Abeta accumulation (over Abeta treatment alone) within the slice culture, as determined by ELISA. However, there are striking differences in the pattern of Abetair among the three isoforms of TGF-beta. Isoforms 1 and 3 produced a cellular pattern of Abeta staining that colocalizes with GS lectin staining (microglia). TGF-beta2 produces dramatic Abeta staining of pyramidal neurons in layers CA1-CA2. In addition to cellular Abeta staining, plaque-like deposits are increased by all of the TGF betas. Although no gross toxicity was observed, morphological neurodegenerative changes were seen in the CA1 region when the slices were treated with Abeta plus TGF-beta2. Our results demonstrate important functional differences among the TGF beta isoforms in their ability to alter the cellular distribution and degradation of Abeta. These changes may be relevant to the pathology of Alzheimer's disease (AD). PMID- 9852576 TI - CREB in the mouse SCN: a molecular interface coding the phase-adjusting stimuli light, glutamate, PACAP, and melatonin for clockwork access. AB - The suprachiasmatic nucleus (SCN) is a central pacemaker in mammals, driving many endogenous circadian rhythms. An important pacemaker target is the regulation of a hormonal message for darkness, the circadian rhythm in melatonin synthesis. The endogenous clock within the SCN is synchronized to environmental light/dark cycles by photic information conveyed via the retinohypothalamic tract (RHT) and by the nocturnal melatonin signal that acts within a feedback loop. We investigated how melatonin intersects with the temporally gated resetting actions of two RHT transmitters, pituitary adenylate cyclase-activating polypeptide (PACAP) and glutamate. We analyzed immunocytochemically the inducible phosphorylation of the transcription factor Ca2+/cAMP response element-binding protein (CREB) in the SCN of a melatonin-proficient (C3H) and a melatonin deficient (C57BL) mouse strain. In vivo, light-induced phase shifts in locomotor activity were consistently accompanied by CREB phosphorylation in the SCN of both strains. However, in the middle of subjective nighttime, light induced larger phase delays in C57BL than in C3H mice. In vitro, PACAP and glutamate induced CREB phosphorylation in the SCN of both mouse strains, with PACAP being more effective during late subjective daytime and glutamate being more effective during subjective nighttime. Melatonin suppressed PACAP- but not glutamate induced phosphorylation of CREB. The distinct temporal domains during which glutamate and PACAP induce CREB phosphorylation imply that during the light/dark transition the SCN switches sensitivity between these two RHT transmitters. Because these temporal domains are not different between C3H and C57BL mice, the sensitivity windows are set independently of the rhythmic melatonin signal. PMID- 9852575 TI - Nitric oxide-producing islet cells modulate the release of sensory neuropeptides in the rat substantia gelatinosa. AB - The substantia gelatinosa of the spinal cord (lamina II) is the major site of integration for nociceptive information. Activation of NMDA glutamate receptor, production of nitric oxide (NO), and enhanced release of substance P and calcitonin gene-related peptide (CGRP) from primary afferents are key events in pain perception and central hyperexcitability. By combining reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry for NO-producing neurons with immunogold labeling for substance P, CGRP, and glutamate, we show that (1) NO-producing neurons in lamina IIi are islet cells; (2) these neurons rarely form synapses onto peptide-immunoreactive profiles; and (3) NADPH diaphorase-positive dendrites are often in close spatial relationship with peptide-containing terminals and are observed at the periphery of type II glomeruli showing glutamate-immunoreactive central endings. By means of confocal fluorescent microscopy in acute spinal cord slices loaded with the Ca2+ indicator Indo-1, we also demonstrate that (1) NMDA evokes a substantial [Ca2+]i increase in a subpopulation of neurons in laminae I-II, with morphological features similar to those of islet cells; (2) a different neuronal population in laminae I IIo, unresponsive to NMDA, displays a significant [Ca2+]i increase after slice perfusion with either substance P and the NO donor 3morpholinosydnonimine (SIN 1); and (3) the responses to both substance P and SIN-1 are either abolished or significantly inhibited by the NK1 receptor antagonist sendide. These results provide compelling evidence that glutamate released at type II glomeruli triggers the production of NO in islet cells within lamina IIi after NMDA receptor activation. The release of substance P from primary afferents triggered by newly synthesized NO may play a crucial role in the cellular mechanism leading to spinal hyperexcitability and increased pain perception. PMID- 9852577 TI - Heteromultimeric delayed-rectifier K+ channels in schwann cells: developmental expression and role in cell proliferation. AB - Schwann cells (SCs) are responsible for myelination of nerve fibers in the peripheral nervous system. Voltage-dependent K+ currents, including inactivating A-type (KA), delayed-rectifier (KD), and inward-rectifier (KIR) K+ channels, constitute the main conductances found in SCs. Physiological studies have shown that KD channels may play an important role in SC proliferation and that they are downregulated in the soma as proliferation ceases and myelination proceeds. Recent studies have begun to address the molecular identity of K+ channels in SCs. Here, we show that a large repertoire of K+ channel alpha subunits of the Shaker (Kv1.1, Kv1.2, Kv1.4, and Kv1.5), Shab (Kv2.1), and Shaw (Kv3.1b and Kv3.2) families is expressed in mouse SCs and sciatic nerve. We characterized heteromultimeric channel complexes that consist of either Kv1.5 and Kv1.2 or Kv1.5 and Kv1.4. In postnatal day 4 (P4) sciatic nerve, most of the Kv1.2 channel subunits are involved in heteromultimeric association with Kv1.5. Despite the presence of Kv1. 1 and Kv1.2 alpha subunits, the K+ currents were unaffected by dendrotoxin I (DTX), suggesting that DTX-sensitive channel complexes do not account substantially for SC KD currents. SC proliferation was found to be potently blocked by quinidine or 4-aminopyridine but not by DTX. Consistent with previous physiological studies, our data show that there is a marked downregulation of all KD channel alpha subunits from P1-P4 to P40 in the sciatic nerve. Our results suggest that KD currents are accounted for by a complex combinatorial activity of distinct K+ channel complexes and confirm that KD channels are involved in SC proliferation. PMID- 9852578 TI - Postsynaptic Ca2+ influx mediated by three different pathways during synaptic transmission at a calyx-type synapse. AB - Whole-cell recordings and Ca2+ flux measurements were made at a giant calyx-type synapse in rat brainstem slices to determine the contribution of glutamate receptor (GluR) channels and voltage-dependent Ca2+ channels (VDCCs) to postsynaptic Ca2+ influx during synaptic transmission. A single presynaptic action potential (AP) evoked an EPSP, followed by a single AP. The EPSP-AP sequence caused a postsynaptic Ca2+ influx of approximately 3.0 pC, primarily through VDCCs ( approximately 70%) and NMDA-type (up to 30%) channels but also through AMPA-type (<5%) GluR channels. At -80 mV, the fractional Ca2+ current (Pf) mediated by AMPA receptor (AMPAR) and NMDA receptor (NMDAR) channels was 1.3 and 11-12%, respectively. Simulations of the time course of Ca2+ influx through GluR channels suggested that the small contribution of AMPAR channels occurred only during the first few milliseconds of an EPSP, whereas influx through NMDAR channels dominated later. The NMDAR-mediated Ca2+ influx was localized in regions covered by the presynaptic terminal, whereas the Ca2+ influx mediated by VDCCs was more homogeneously distributed. Because of the temporal and spatial differences, calcium ions entering through the three different pathways are likely to activate different intracellular targets in the postsynaptic cell. PMID- 9852579 TI - p21 ras and phosphatidylinositol-3 kinase are required for survival of wild-type and NF1 mutant sensory neurons. AB - Nerve growth factor (NGF) is a required differentiation and survival factor for sympathetic and a majority of neural crest-derived sensory neurons in the developing vertebrate peripheral nervous system. Although much is known about the function of NGF, the intracellular signaling cascade that it uses continues to be a subject of intense study. p21 ras signaling is considered necessary for sensory neuron survival. How additional intermediates downstream or in parallel may function has not been fully understood yet. Two intracellular signaling cascades, extra cellular regulated kinase (erk) and phosphatidylinositol-3 (PI 3) kinase, transduce NGF signaling in the pheochromocytoma cell line PC12. To elucidate the role these cascades play in survival and differentiation, we used a combination of recombinant adenoviruses and chemical inhibitors to perturb these pathways in sensory neurons from wild-type mice and mice deficient for neurofibromin in which the survival and differentiation pathway is constitutively active. We demonstrate that ras activity is both necessary and sufficient for the survival of embryonic sensory neurons. Downstream of ras, however, the erk cascade is neither required nor sufficient for neuron survival or overall differentiation. Instead, the activity of PI 3 kinase is necessary for the survival of the wild-type and neurofibromin-deficient neurons. Therefore, we conclude that in sensory neurons, NGF acts via a signaling pathway, which includes both ras and PI 3 kinase. PMID- 9852580 TI - Neurite outgrowth stimulated by neural cell adhesion molecules requires growth associated protein-43 (GAP-43) function and is associated with GAP-43 phosphorylation in growth cones. AB - The mechanisms whereby cell adhesion molecules (CAMs) promote axonal growth and synaptic plasticity are poorly understood. Here we show that the neurite outgrowth stimulated by NCAM-mediated fibroblast growth factor (FGF) receptor activation in cerebellar granule cells is associated with increased GAP-43 phosphorylation on serine-41. In contrast, neither NCAM nor FGF was able to stimulate neurite outgrowth in similar neurons from mice in which the GAP-43 gene had been deleted by homologous recombination. Integrin-mediated neurite outgrowth was unaffected by GAP-43 deletion. Both neurite outgrowth and rapid phosphorylation of GAP-43 in isolated growth cones required the first three Ig domains of a NCAM-Fc chimera and were stimulated maximally at 5 micrograms/ml (approximately 50 nM). Likewise, GAP-43 phosphorylation in isolated growth cones also was stimulated by an L1-Fc chimera. Both neurite outgrowth and NCAM stimulated GAP-43 phosphorylation were inhibited by antibodies to the FGF receptor and a diacylglycerol lipase inhibitor (RHC80267) that blocks the production of arachidonic acid in response to activation of the FGF receptor. Direct activation of the FGF receptor and the arachidonic acid cascade with either basic FGF or melittin also resulted in increased GAP-43 phosphorylation. These data suggest that the stimulation of neurite outgrowth by NCAM requires GAP 43 function and that GAP-43 phosphorylation in isolated growth cones occurs via an FGF receptor-dependent increase in arachidonic acid. PMID- 9852581 TI - UNC-55, an orphan nuclear hormone receptor, orchestrates synaptic specificity among two classes of motor neurons in Caenorhabditis elegans. AB - Loss of UNC-55 function in the nematode Caenorhabditis elegans causes one motor neuron class, the ventral D (VD) motor neurons, to adopt the synaptic pattern of another motor neuron class, the dorsal D (DD) motor neurons. Here we show that unc-55 encodes a member of the nuclear hormone receptor gene family that is similar to the vertebrate chicken ovalbumin upstream promoter transcription factors. Although the VD and DD motor neuron classes arise from different lineages at different developmental stages, they share a number of structural and functional features that appear to be the product of identical genetic programs. UNC-55 is expressed in the VD but not the DD motor neurons to modify this genetic program and to create the synaptic pattern that distinguishes the two motor neuron classes from one another. PMID- 9852582 TI - Chronic interleukin-6 alters NMDA receptor-mediated membrane responses and enhances neurotoxicity in developing CNS neurons. AB - Recent studies show that the cytokine interleukin-6 (IL-6) is expressed at elevated levels in the CNS in several disease states and contributes to the neuropathological process. The mechanisms through which IL-6 exerts its CNS effects are primarily unknown. We have investigated the pathophysiological effects of IL-6 on developing CNS neurons using a culture model system and a chronic treatment paradigm. Here, we show, using current- and voltage-clamp recordings, that chronic IL-6 treatment of developing cerebellar granule neurons increases the membrane and current response to NMDA and that these effects are the primary mechanism through which IL-6 produces an enhanced calcium signal to NMDA. We also show that calcium influx through voltage-sensitive calcium channels contributes to the enhanced calcium signal to NMDA in the IL-6-treated neurons in a developmentally regulated manner and that the membrane depolarization to NMDA is more sensitive to the NMDA receptor antagonist ifenprodil in the IL-6-treated neurons compared with control neurons at a late developmental stage, consistent with a larger proportion of NMDA receptors containing the NMDAR2B subunit in the IL-6-treated neurons. Additional studies show that IL-6 treatment reduces the number of granule neurons in culture and enhances neurotoxicity involving NMDA receptors. These results support a pathological role for IL-6 in the CNS and indicate that NMDA receptor-mediated functions are likely to play a critical role in neuropathological changes observed in CNS diseases associated with elevated CNS levels of IL-6. PMID- 9852583 TI - The chemokine growth-regulated oncogene-alpha promotes spinal cord oligodendrocyte precursor proliferation. AB - Chemokines, (chemotactic cytokines) are a family of regulatory molecules involved in modulating inflammatory responses. Here we demonstrate that the chemokine growth-regulated oncogene-alpha (GRO-alpha) is a potent promoter of oligodendrocyte precursor proliferation. The proliferative response of immature spinal cord oligodendrocyte precursors to their major mitogen, platelet derived growth factor (PDGF), is dramatically enhanced by GRO-alpha present in spinal cord conditioned medium. One source of GRO-alpha is a subset of spinal cord astrocytes. Cultures of astrocytes contain GRO-alpha mRNA and protein and secrete biologically active concentrations of GRO-alpha. In postnatal spinal cord white matter the location of GRO-alpha-immunoreactive cells is developmentally regulated: GRO-alpha+ cells first appear in ventral and later in dorsal spinal cord white matter. These results suggest that localized proliferation of oligodendrocytes is mediated by synergy between PDGF and GRO-alpha. PMID- 9852584 TI - Synaptic modifications in cultured hippocampal neurons: dependence on spike timing, synaptic strength, and postsynaptic cell type. AB - In cultures of dissociated rat hippocampal neurons, persistent potentiation and depression of glutamatergic synapses were induced by correlated spiking of presynaptic and postsynaptic neurons. The relative timing between the presynaptic and postsynaptic spiking determined the direction and the extent of synaptic changes. Repetitive postsynaptic spiking within a time window of 20 msec after presynaptic activation resulted in long-term potentiation (LTP), whereas postsynaptic spiking within a window of 20 msec before the repetitive presynaptic activation led to long-term depression (LTD). Significant LTP occurred only at synapses with relatively low initial strength, whereas the extent of LTD did not show obvious dependence on the initial synaptic strength. Both LTP and LTD depended on the activation of NMDA receptors and were absent in cases in which the postsynaptic neurons were GABAergic in nature. Blockade of L-type calcium channels with nimodipine abolished the induction of LTD and reduced the extent of LTP. These results underscore the importance of precise spike timing, synaptic strength, and postsynaptic cell type in the activity-induced modification of central synapses and suggest that Hebb's rule may need to incorporate a quantitative consideration of spike timing that reflects the narrow and asymmetric window for the induction of synaptic modification. PMID- 9852585 TI - Segmental specificity of chick sympathetic preganglionic projections is influenced by preganglionic neurons from neighboring spinal cord segments. AB - Sympathetic preganglionic neurons of the chick are located between the brachial and lumbosacral enlargements of the spinal cord. Their axons exit the spinal cord via their adjacent ventral roots and project rostrally or caudally along the sympathetic trunk to innervate sympathetic ganglia. The projections of sympathetic preganglionic neurons are segmentally specific. Neurons from the 16th cervical (C16) and the first thoracic (T1) spinal cord segments project predominantly in the rostral direction, whereas those from the fifth thoracic (T5) to the first lumbar (L1) spinal segments project predominantly in the caudal direction. Neurons from intervening spinal cord segments (T2-T4) project in rostral and caudal directions. In the present study, neural tube manipulations show that the direction of preganglionic projections is altered by both the elimination and addition of preganglionic neurons projecting into the sympathetic trunk from neighboring segments. The present study also compares the projections of preganglionic neurons from transplants of multiple neural tube segments with those from transplants of single neural tube segments reported in a previous study (Yip, 1987). In the previous study when single thoracic neural tube segments were transplanted to the cervical level, preganglionic neurons did not maintain their original projection patterns. The present study found that, when contiguous neighboring segments were transplanted to the cervical level, preganglionic neurons maintained projection patterns characteristic of their original segmental levels. These results indicate that the direction of preganglionic projections can be influenced by neurons from neighboring segments, suggesting that the formation of segmentally specific preganglionic projections during embryogenesis may involve the interactions of preganglionic neurons with those from neighboring spinal cord segments. PMID- 9852586 TI - Expression pattern and neurotrophic role of the c-fms proto-oncogene M-CSF receptor in rodent Purkinje cells. AB - To investigate whether the c-fms proto-oncogene plays a role in the CNS, we examined its expression in mouse brain. We found that c-fms-positive Purkinje cells first appeared in caudal cerebellum at postnatal day 0 (P0) arranged in a parasagittal manner, and most Purkinje cells gradually became positive by P6. This differential expression was not seen from P7 to adulthood, and the parasagittal pattern until P5 was different from those of L7, zebrins, and the integrin beta1 subunit. No neuronal expression of c-fms was found in the other brain regions examined. In both reeler and weaver mutant mice in the adult stage, all Purkinje cells were positive for c-fms as in the wild-type controls; however, the parasagittal bands of c-fms-positive Purkinje cells were observed even in the adult staggerer mutant. To check the neurotrophic effect of macrophage colony stimulating factor (M-CSF), we immunostained cerebella derived from osteopetrotic mutant mice, that is, those devoid of active M-CSF. We found that the number of calbindin-positive Purkinje cells in a given cerebellum began to decrease substantially during the initial 4-5 weeks of the postnatal period. In addition, cultured Purkinje cells were dependent on M-CSF for their survival. These data suggest that expression of the c-fms gene is intrinsically programmed in the Purkinje cells and never affected by the afferent synaptic input and that neuronal survival of Purkinje cells is dependent on M-CSF after weaning. Therefore, c-fms is considered to be a new developmental marker for Purkinje cells. PMID- 9852587 TI - The role of nitric oxide and NMDA receptors in the development of motor neuron dendrites. AB - Nitric oxide (NO) has been implicated in the establishment of precise synaptic connectivity throughout the neuroaxis in several species. To determine the contribution of NO to NMDA receptor-dependent dendritic growth in motor neurons, we administered the NMDA antagonist MK-801 to wild-type mice and neuronal nitric oxide synthase (nNOS) knock-out mice between postnatal days 7 and 14. Compared to saline-treated wild-type animals the number of dendritic bifurcations was significantly reduced in nNOS knock-out animals and MK-801-treated wild-type animals. There was no significant difference in dendritic bifurcation between MK 801-treated wild-type, MK-801-treated nNOS knock-out, and saline-treated nNOS knock-out animals, suggesting that nNOS knock-out and NMDA receptor block had similar effects. The path of the longest dendrite and the number of primary dendrites was the same in all treatment groups, indicating an effect specific to bifurcation. Sholl analysis revealed that differences in bifurcation numbers occurred between 160 and 320 micrometers from the cell body, the distance at which second, third, and fourth order dendrites are most prevalent. Dendrite order analyses confirmed a significant reduction in numbers, but not lengths, of third and fourth order dendrites in nNOS knock-out and drug-treatment groups. Finally, immunohistochemical examination of the developing spinal cord indicated that NMDA receptors and nNOS are colocalized within interneurons surrounding the motor neuron pool. These results support the view that at least part of NMDA receptor-dependent arborization of motor neuron dendrites is mediated by the local production of NO within the developing spinal cord. PMID- 9852588 TI - Randomized retinal ganglion cell axon routing at the optic chiasm of GAP-43 deficient mice: association with midline recrossing and lack of normal ipsilateral axon turning. AB - During mammalian development, retinal ganglion cell (RGC) axons from nasal retina cross the optic chiasm midline, whereas temporal retina axons do not and grow ipsilaterally, resulting in a projection of part of the visual world onto one side of the brain while the remaining part is represented on the opposite side. Previous studies have shown that RGC axons in GAP-43-deficient mice initially fail to grow from the optic chiasm to form optic tracts and are delayed temporarily in the midline region. Here we show that this delayed RGC axon exit from the chiasm is characterized by abnormal randomized axon routing into the ipsilateral and contralateral optic tracts, leading to duplicated representations of the visual world in both sides of the brain. Within the chiasm, individual contralaterally projecting axons grow in unusual semicircular trajectories, and the normal ipsilateral turning of ventral temporal axons is absent. These effects on both axon populations suggest that GAP-43 does not mediate pathfinding specifically for one or the other axon population but is more consistent with a model in which the initial pathfinding defect at the chiasm/tract transition zone leads to axons backing up into the chiasm, resulting in circular trajectories and eventual random axon exit into one or the other optic tract. Unusual RGC axon trajectories include chiasm midline recrossing similar to abnormal CNS midline recrossing in invertebrate "roundabout" mutants and Drosophila with altered calmodulin function. This resemblance and the fact that GAP-43 also has been proposed to regulate calmodulin availability raise the possibility that calmodulin function is involved in CNS midline axon guidance in both vertebrates and invertebrates. PMID- 9852589 TI - Regeneration of adult rat corticospinal axons induced by transplanted olfactory ensheathing cells. AB - Precisely localized focal stereotaxic electrolytic lesions were made in the corticospinal tract at the level of the first to second cervical segments in the adult rat. This consistently destroyed all central nervous tissue elements (axons, astrocytes, oligodendrocytes, microglia, and microvessels) in a highly circumscribed area. In a group of these rats immediately after lesioning, a suspension of cultured adult olfactory ensheathing cells was transplanted into the lesion site. Within the first week after transplantation, the cut corticospinal axons (identified by anterograde transport of biotin dextran) extended caudally along the axis of the corticospinal tract as single, fine, minimally branched sprouts that ended in a simple tip, often preceded by a small varicosity. By 3 weeks, the regenerating axons, ensheathed by P0-positive peripheral myelin had accumulated into parallel bundles, which now extended across the full length of the lesioned area and reentered the caudal part of the host corticospinal tract. The transplants contained two main types of cells: (1) p75-expressing S cells, which later formed typical peripheral one-to-one myelin sheaths around individual ensheathed axons, and (2) fibronectin-expressing A cells, which aggregated into tubular sheaths enclosing bundles of myelinated axons. The point of reentry of the axons into the central nervous territory of the caudal host corticospinal tract was marked by the resumption of oligodendrocytic myelination. Thus the effect of the transplant was to form a "patch" of peripheral-type tissue across which the cut central axons regenerated and then continued to grow along their original central pathway. PMID- 9852590 TI - The connection from cortical area V1 to V5: a light and electron microscopic study. AB - Area V5 (middle temporal) in the superior temporal sulcus of macaque receives a direct projection from the primary visual cortex (V1). By injecting anterograde tracers (biotinylated dextran and Phaseolus vulgaris lectin) into V1, we have examined the synaptic boutons that they form in V5 in the electron microscope. Nearly 80% of the target cells in V5 were spiny (excitatory). The boutons formed asymmetric (Gray's type 1) synapses with spines (54%), dendrites (33%), and somata (13%). All somatic targets and some (26%) of the target dendritic shafts showed features characteristic of smooth (inhibitory) cells. Each bouton formed, on average, 1.7 synapses. The larger boutons formed multiple synapses with the same neuron and completely enveloped the entire spine head. On most dendritic shafts and all somata the postsynaptic density en face was disk-shaped but in about half the cases the reconstructed postsynaptic densities of synapses on spines appeared as complete or partial annuli. Even in the zones of densest innervation only 3% of the asymmetric synapses were formed by the labeled boutons. Although the V1 projection forms only a small minority of synapses in V5, its affect could be considerably amplified by local circuits in V5, in a way analogous to the amplification of the small thalamic input to area V1. PMID- 9852591 TI - A role for transforming growth factor alpha as an inducer of astrogliosis. AB - TGFalpha is a member of the epidermal growth factor (EGF) family with which it shares the same receptor, the EGF receptor (EGFR). Synthesis of TGFalpha and EGFR in reactive astrocytes developing after CNS insults is associated with the differentiative and mitogenic effects of TGFalpha on cultured astrocytes. This suggests a role for TGFalpha in the development of astrogliosis. We evaluated this hypothesis using transgenic mice bearing the human TGFalpha cDNA under the control of the zinc-inducible metallothionein promoter. Expression levels of glial fibrillary acidic protein (GFAP) and vimentin and morphological features of astrocytes were used as indices of astroglial reactivity in adult transgenic versus wild-type mice provided with ZnCl2 in their water for 3 weeks. In the striatum, the hippocampus, and the cervical spinal cord, the three CNS areas monitored, transgenic mice displayed enhanced GFAP mRNA and protein levels and elevated vimentin protein levels. GFAP-immunoreactive astrocytes exhibited numerous thick processes and hypertrophied somata, which are characteristic aspects of reactive astrocytes. Their number increased additionally in the striatum and the spinal cord, but no astrocytic proliferation was observed using bromodeoxyuridine immunohistochemistry. Neither the morphology nor the number of microglial cells appeared modified. A twofold increase in phosphorylated EGFR was detected in the striatum and was associated with the immunohistochemical detection of numerous GFAP-positive astrocytes bearing the EGFR, suggesting a direct action of TGFalpha on astrocytes. Altogether, these results demonstrate that enhanced TGFalpha synthesis is sufficient to trigger astrogliosis throughout the CNS, whereas microglial metabolism is unaffected. PMID- 9852592 TI - D1 receptor in interneurons of macaque prefrontal cortex: distribution and subcellular localization. AB - Working memory performance is influenced by dopamine activation of D1 family dopamine receptors in the prefrontal cortex; working memory performance is maximal at moderate stimulation of D1 family receptors and is reduced by either higher or lower levels of D1 stimulation. The neuronal mechanisms that underlie this complex relationship are not yet understood. Previous work from this laboratory has demonstrated that the D1 family receptors, D1 and D5, are located in different compartments of pyramidal cells. Here we use an antibody specific to the D1 receptor and double-label immunohistochemistry at the light and electron microscopic level to demonstrate that D1-like immunoreactivity (D1-LIR) is also present in interneurons. D1 receptor is prevalent in parvalbumin-containing interneurons and is less common in calretinin-containing interneurons. At the ultrastructural level, D1-LIR is found associated with the Golgi apparatus and endoplasmic reticulum in the soma, with the membranes of vesicles in proximal dendrites, and with the plasma membrane on distal dendrites, where it is often located near asymmetric synapses. In addition, D1-LIR is also seen in presynaptic axon terminals, which give rise to symmetric synapses onto dendritic shafts and soma. These results raise the possibility that the circuit basis of working memory in the prefrontal cortex involves a D1-mediated inhibitory component. PMID- 9852593 TI - Dopamine modulates the responsivity of mediodorsal thalamic cells recorded in vitro. AB - The mediodorsal thalamic nucleus (MD) receives convergent inputs from subcortical limbic structures that overlap with a dopaminergic (DA) innervation. In this study, we describe the effects of DA agonists on the basal and evoked electrophysiological activity of identified thalamic cells of rats recorded in vitro. Administration of the D1 agonist SFK 38393 (10 microM) did not produce a clear effect on the physiological properties of the thalamic cells recorded. In contrast, bath administration of the D2 agonist quinpirole (10 microM) resulted in an enhancement of membrane excitability, facilitation of the occurrence of low threshold spikes (LTSs), and changes in the resting membrane potential of the thalamic cells tested. The quinpirole-mediated responses were reversed by administration of the D2 antagonist haloperidol. Results from experiments performed with different [K+] and K+ channel blockers suggest that the effects of quinpirole are mediated at least in part by changes in K+ conductances. The results from this study suggest that DA can modulate the excitability of thalamic cells and in turn may influence the way that the thalamocortical system integrates information. PMID- 9852595 TI - Neurotransmitter coupling through gap junctions in the retina. AB - Although all bipolar cells in the retina probably use the excitatory transmitter glutamate, approximately half of the cone bipolar cells also contain elevated levels of the inhibitory transmitter glycine. Some types of cone bipolar cells make heterologous gap junctions with rod amacrine cells, which contain elevated levels of glycine, leading to the hypothesis that the bipolar cells obtain their glycine from amacrine cells. Experimental support for this hypothesis is now provided by three independent lines of evidence. First, the glycine transporter GLYT1 is expressed by the glycine-containing amacrine cells but not by the glycine-containing bipolar cells, suggesting that only the amacrine cells are functionally glycinergic. Second, the gap-junction blocker carbenoxolone greatly reduces exogenous 3H-glycine accumulation into the bipolar cells but not the amacrine cells. Moreover, when the endogenous glycine stores in both cell classes are depleted by incubating the retina with a glycine-uptake inhibitor, carbenoxolone blocks the subsequent glycine replenishment of the bipolar cells but not the amacrine cells. Third, intracellular injection of rod amacrine cells with the gap-junction permeant tracer Neurobiotin secondarily labels a heterogenous population of cone bipolar cells, all of which show glycine immunoreactivity. Taken together, these findings indicate that the elevated glycine in cone bipolar cells is not derived by high-affinity uptake or de novo synthesis but is obtained by neurotransmitter coupling through gap junctions with glycinergic amacrine cells. Thus transmitter content may be an unreliable indicator of transmitter function for neurons that make heterologous gap junctions. PMID- 9852594 TI - Amphetamine-induced behavior, dopamine release, and c-fos mRNA expression: modulation by environmental novelty. AB - We have shown recently that the psychomotor activating effects of amphetamine in the rat are much greater when this drug is administered in association with environmental novelty than when it is given in a home environment. The main purpose of the present study was to explore the neural basis of this phenomenon. We found, using in situ hybridization of c-fos mRNA, that the pattern of neuronal activation in the cortex, in the caudate, in the shell and core of the nucleus accumbens, and in other subcortical structures was markedly different when amphetamine (2.0 mg/kg, i.p.) was given in association with exposure to environmental novelty relative to when it was given at home. In most brain regions the magnitude of c-fos expression was over two times greater in rats given amphetamine plus novelty than in rats given amphetamine alone. In contrast, an in vivo microdialysis study indicated that environmental novelty did not affect amphetamine-induced dopamine release in either caudate or nucleus accumbens. Furthermore, a unilateral 6-hydroxydopamine lesion of the mesostriatal dopamine system reduced amphetamine- but not novelty-induced c-fos expression. Finally, we found no differences in the amount of corticosterone secreted after exposure to novelty, amphetamine, or both, suggesting that corticosterone does not play a critical role in the ability of novelty to modulate amphetamine induced psychomotor activation. In conclusion, it seems that environmental novelty alters the neurobiological effects of amphetamine independently of the primary neuropharmacological actions of this drug in the striatum. PMID- 9852596 TI - Anatomic evidence of a three-dimensional mosaic pattern of tonotopic organization in the ventral complex of the lateral lemniscus in cat. AB - The ventral complex of the lateral lemniscus (VCLL, i.e., the ventral and intermediate nuclei) is composed of cells embedded in the fibers of the lateral lemniscus. These cells are involved in the processing of monaural information and receive input from the collaterals of the fibers ascending to the inferior colliculus. Whereas tonotopic organization is a feature of all other nuclei of the auditory system, this functional principle is debated in the VCLL. We have made focal injections of the tracer biotinylated dextran amine into different frequency band representations of the inferior colliculus in cat. Retrogradely labeled cells and terminal fibers (collaterals of efferent local axons and other ascending lemniscal fibers) were found in the ipsilateral VCLL. The spatial distribution of the labeling was analyzed using three-dimensional (3-D) reconstruction and computer graphical visualization techniques. A complex topographic organization was found. In all cases, labeled fibers and cells were distributed in multiple clusters throughout the dorsoventral extent of the VCLL. The shape, size, and location of the labeled clusters suggest an interdigitation of clusters assigned to different frequency-band representations. But an overall mediolateral distribution gradient was observed, with high frequencies represented medially and lower frequencies progressively more laterally. We conclude that the clusters may represent discontinuous frequency-band compartments as a counterpart to the continuous laminar compartments in the remaining auditory nuclei. The 3-D orderly mosaic pattern indicates that the VCLL preserves the spectral decomposition originated in the cochlea in a way that facilitates across-frequency integration. PMID- 9852597 TI - Noradrenergic excitation of magnocellular neurons in the rat hypothalamic paraventricular nucleus via intranuclear glutamatergic circuits. AB - Noradrenergic projections to the hypothalamus play a critical role in the afferent control of oxytocin and vasopressin release. Recent evidence for intrahypothalamic glutamatergic circuits prompted us to test the hypothesis that the excitatory effect of noradrenergic inputs on oxytocin and vasopressin release is mediated in part by local glutamatergic interneurons. The voltage response to norepinephrine (30-300 microM) was tested with whole-cell recordings in putative magnocellular neurons of the paraventricular nucleus (PVN) in hypothalamic slices (400 micrometers). Norepinephrine elicited an alpha1 receptor-mediated direct depolarization in 23% of the magnocellular neurons tested; however, the most prominent response, seen in 42% of the magnocellular neurons, was an alpha1 receptor-mediated increase in the frequency of EPSPs. The norepinephrine-induced increase in EPSPs was blocked by tetrodotoxin and by ionotropic glutamate receptor antagonists, suggesting that norepinephrine excited presynaptic glutamate neurons to cause an increase in spike-mediated transmitter release. The increase in EPSPs also was observed in a surgically isolated PVN preparation (64% of cells) and with microdrop applications of norepinephrine (1 mM, 33% of cells) and glutamate (0.5-1 mM, 28%) in the PVN, indicating that the norepinephrine sensitive presynaptic glutamate neurons are located within the PVN. Biocytin injection and subsequent immunohistochemical labeling revealed that both oxytocin and vasopressin neurons responded to norepinephrine. Our data indicate that magnocellular neurons of the PVN receive excitatory inputs from intranuclear glutamatergic neurons that express alpha1-adrenoreceptors. These glutamatergic interneurons may serve as an excitatory relay in the afferent noradrenergic control of oxytocin and vasopressin release under certain physiological conditions. PMID- 9852598 TI - Extraocular motor unit and whole-muscle responses in the lateral rectus muscle of the squirrel monkey. AB - Because primate studies provide data for the current experimental models of the human oculomotor system, we investigated the relationship of lateral rectus muscle motoneuron firing to muscle unit contractile characteristics in the squirrel monkey. Also examined was the correlation of whole-muscle contractile force with the degree of evoked eye displacement. A force transducer was used to record lateral rectus whole-muscle or muscle unit contraction in response to abducens whole-nerve stimulation or stimulation of single abducens motoneurons or axons. Horizontal eye displacement was recorded using a magnetic search coil. (1) Motor units could be categorized based on contraction speed (fusion frequency) and fatigue. (2) The kt value (change in motoneuronal firing necessary to increase motor unit force by 1.0 mg) of the units correlated with maximum tetanic tension. (3) There was some tendency for maximum tetanic tension of this unit population to separate into three groups. (4) At a constant frequency of 100 Hz, 95% of the motor units demonstrated significantly different force levels dependent on immediately previous stimulation history (hysteresis). (5) A mean force change of 0.32 gm/ degrees and a mean frequency change of 4.7 Hz/ degrees of eye displacement were observed in response to whole-nerve stimulation. These quantitative data provide the first contractile measures of primate extraocular motor units. Models of eye movement dynamics may need to consider the nonlinear transformations observed between stimulation rate and muscle tension as well as the probability that as few as two to three motor units can deviate the eye 1 degrees. PMID- 9852599 TI - Selective neuronal expression of green fluorescent protein with cytomegalovirus promoter reveals entire neuronal arbor in transgenic mice. AB - In simple nervous systems, identified groups of neurons can be studied in depth. To allow the same advantage in the mammalian brain, we have generated green fluorescent protein (GFP) transgenic mice in which only a few types of neurons are strongly labeled with a fluorescent molecule, which the neurons synthesize internally, allowing the cells, their dendrites, filopodia, and axons to be identified in both living and fixed CNS, in slices and culture. The same neurons, with GFP expression controlled by part of the major immediate early promoter of human cytomegalovirus (CMV), show GFP beginning early in development, from one generation to the next, allowing cellular and physiological studies of axonal and dendritic growth, fate mapping, anatomical connections, and synapse formation in identified neurons. The human CMV promoter sequence we used was different from that used in previous work with other reporter genes and gave a dramatically different pattern of expression. Two transgenic lines with the same CMV promoter show similar anatomical patterns of expression in the present study. Strong GFP labeling was found in a subpopulation of mossy fibers that innervated parasagittal bands in the cerebellar cortex and olfactory axons that projected into the olfactory bulb, subsets of motoneurons and dorsal root ganglion cells, granule but not mitral cells of the olfactory bulb, and a group of neurons in the hypothalamic suprachiasmatic nucleus. A novel type of neuron was strongly labeled in the olfactory bulb external plexiform layer. In normal brains, CMV does not constitute a threat, but in the developing brain, CMV can cause debilitating neurodegeneration and death; studies using the CMV promoter aid in understanding the affinity of CMV that has been suggested for specific brain regions. PMID- 9852600 TI - Calcium currents of rhythmic neurons recorded in the isolated respiratory network of neonatal mice. AB - To obtain a quantitative characterization of voltage-activated calcium currents in respiratory neurons, we performed voltage-clamp recordings in the transverse brainstem slice of mice from neurons located within the ventral respiratory group. It is assumed that this medullary region contains the neuronal network responsible for generating the respiratory rhythm. This study represents one of the first attempts to analyze quantitatively the currents in respiratory neurons. The inward calcium currents of VRG neurons consisted of two components: a high voltage-activated (HVA) and a low voltage-activated (LVA) calcium current. The activation threshold of the HVA current was at -40 mV. It was fully activated (peak voltage) between -10 and 0 mV. The half-maximal activation (V50) was at 27. 29 mV +/- 1.15 (n = 24). The HVA current was inactivated completely at a holding potential of -35 mV and fully deinactivated at a holding potential of -65 mV (V50, -52.26 mV +/- 0.27; n = 18). The threshold for the activation of the LVA current was at -65 mV. This current had its peak voltage between -50 and -40 mV (mean, V50 = -59. 15 mV +/- 0.21; n = 15). The LVA current was inactivated completely at a holding potential of -65 mV and deinactivated fully at a holding potential of -95 mV (mean, V50 = -82.40 mV +/- 0.32; n = 38). These properties are consistent with other studies suggesting that the LVA current is a T-type current. The properties of these inward currents are discussed with respect to their role in generating Ca2+ potentials that may contribute to the generation of the mammalian respiratory rhythm. PMID- 9852601 TI - Suppression of ethanol-reinforced behavior by naltrexone is associated with attenuation of the ethanol-induced increase in dialysate dopamine levels in the nucleus accumbens. AB - The opiate antagonist naltrexone suppresses ethanol-reinforced behavior in animals and decreases ethanol intake in humans. However, the mechanisms underlying these actions are not well understood. Experiments were designed to test the hypothesis that naltrexone attenuates the rewarding properties of ethanol by interfering with ethanol-induced stimulation of dopamine activity in the nucleus accumbens (NAcc). Simultaneous measures of the effects of naltrexone on dialysate dopamine levels in the NAcc and on operant responding for oral ethanol were used. Male Wistar rats were trained to self-administer ethanol (10 15%, w/v) in 0.2% (w/v) saccharin during daily 30 min sessions and were surgically prepared for intracranial microdialysis. Experiments began after reliable self-administration was established. Rats were injected with naltrexone (0.25 mg/kg, s.c.) or saline and 10 min later were placed inside the operant chamber for a 20 min waiting period with no ethanol available, followed by 30 min of access to ethanol. A transient rise in dialysate dopamine levels was observed during the waiting period, and this effect was not altered by naltrexone. Ethanol self-administration reliably increased dopamine levels in controls. Naltrexone significantly suppressed ethanol self-administration and prevented ethanol induced increases in dialysate dopamine levels. Subsequent dose-effect analyses established that the latter effect was not merely a function of reduced ethanol intake but that naltrexone attenuated the efficacy of ethanol to elevate dialysate dopamine levels. These results suggest that suppression of ethanol self administration by opiate antagonists is the result of interference with dopamine dependent aspects of ethanol reinforcement, although possible additional effects via nondopaminergic mechanisms cannot be eliminated as a factor in opiate antagonist-induced reduction of ethanol intake. PMID- 9852602 TI - Mechanisms of action and targets of nitric oxide in the oculomotor system. AB - Nitric oxide (NO) production by neurons in the prepositus hypoglossi (PH) nucleus is necessary for the normal performance of eye movements in alert animals. In this study, the mechanism(s) of action of NO in the oculomotor system has been investigated. Spontaneous and vestibularly induced eye movements were recorded in alert cats before and after microinjections in the PH nucleus of drugs affecting the NO-cGMP pathway. The cellular sources and targets of NO were also studied by immunohistochemical detection of neuronal NO synthase (NOS) and NO-sensitive guanylyl cyclase, respectively. Injections of NOS inhibitors produced alterations of eye velocity, but not of eye position, for both spontaneous and vestibularly induced eye movements, suggesting that NO produced by PH neurons is involved in the processing of velocity signals but not in the eye position generation. The effect of neuronal NO is probably exerted on a rich cGMP-producing neuropil dorsal to the nitrergic somas in the PH nucleus. On the other hand, local injections of NO donors or 8-Br-cGMP produced alterations of eye velocity during both spontaneous eye movements and vestibulo-ocular reflex (VOR), as well as changes in eye position generation exclusively during spontaneous eye movements. The target of this additional effect of exogenous NO is probably a well defined group of NO-sensitive cGMP-producing neurons located between the PH and the medial vestibular nuclei. These cells could be involved in the generation of eye position signals during spontaneous eye movements but not during the VOR. PMID- 9852603 TI - Psychostimulant-induced Fos protein expression in the thalamic paraventricular nucleus. AB - Lesions of glutamatergic afferents to the nucleus accumbens have been reported to block psychostimulant-induced behavioral sensitization. However, thalamic glutamatergic projections to the nucleus accumbens have received little attention in the context of psychostimulant actions. We examined the effects of acute amphetamine and cocaine administration on expression of Fos protein in the thalamic paraventricular nucleus (PVT), which provides glutamatergic inputs to the nucleus accumbens and also receives dopaminergic afferents. Immunoblot and immunohistochemical studies revealed that both psychostimulants dose-dependently increased PVT Fos expression. PVT neurons retrogradely labeled from the nucleus accumbens were among the PVT cells that showed a Fos response to amphetamine. D2 family dopamine agonists, including low doses of the D3-preferring agonist 7-OH DPAT, increased the numbers of Fos-like-immunoreactive neurons in the PVT. Conversely, the effects of cocaine and amphetamine on PVT Fos expression were blocked by pretreatment with the dopamine D2/3 antagonist raclopride. Because PVT neurons express D3 but not other dopamine receptor transcripts, it appears that psychostimulants induce Fos in PVT neurons through a D3 dopamine receptor. We suggest that the PVT may be an important part of an extended circuit subserving both the arousing properties and reinforcing aspects of psychostimulants. PMID- 9852604 TI - An oscillatory short-term memory buffer model can account for data on the Sternberg task. AB - A limited number (7 +/- 2) of items can be held in human short-term memory (STM). We have previously suggested that observed dual (theta and gamma) oscillations could underlie a multiplexing mechanism that enables a single network to actively store up to seven memories. Here we have asked whether models of this kind can account for the data on the Sternberg task, the most quantitative measurements of memory search available. We have found several variants of the oscillatory search model that account for the quantitative dependence of the reaction time distribution on the number of items (S) held in STM. The models differ on the issues of (1) whether theta frequency varies with S and (2) whether the phase of ongoing oscillations is reset by the probe. Using these models the frequencies of dual oscillations can be derived from psychophysical data. The derived values (ftheta = 6-10 Hz; fgamma = 45-60 Hz) are in reasonable agreement with experimental values. The exhaustive nature of the serial search that has been inferred from psychophysical measurements can be plausibly explained by these oscillatory models. One argument against exhaustive serial search has been the existence of serial position effects. We find that these effects can be explained by short-term repetition priming in the context of serial scanning models. Our results strengthen the case for serial processing and point to experiments that discriminate between variants of the serial scanning process. PMID- 9852605 TI - Differential regulation of neurotrophin and trk receptor mRNAs in catecholaminergic nuclei during chronic opiate treatment and withdrawal. AB - The neurotrophins brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT 3) and their receptors trkB and trkC, respectively, are expressed in the locus coeruleus (LC) and ventral tegmental area (VTA), brain regions known to be involved in opiate addiction. Previously, administration of exogenous neurotrophins has been shown to oppose effects of chronic morphine treatment on LC and VTA neurons. However, the response of endogenous neurotrophins in LC and VTA to opiate treatment is unknown. In this study, BDNF, NT-3, trkB, and trkC mRNAs were analyzed in these regions after chronic morphine treatment and during antagonist precipitated withdrawal. Although chronic morphine exposure resulted in only modest increases in BDNF and NT-3 mRNA expression in LC, precipitated withdrawal led to a marked, rapid, and prolonged increase in BDNF mRNA and a delayed decrease in NT-3 mRNA. Levels of trkB and trkC mRNAs, which were unchanged by chronic morphine treatment, were elevated in LC at 2 and 6 hr of withdrawal. By 20 hr, trkB mRNA levels in LC had returned to control, whereas trkC mRNA levels fell below control values. In contrast to the substantial alterations observed in LC, there was no regulation of the neurotrophins or trk mRNAs within the VTA during chronic opiate treatment or withdrawal, with the exception of an increase in trkB mRNA at 6 hr of withdrawal. These results suggest that neurotrophins and their receptors per se may be involved in opiate induced plasticity of the LC, whereas other mechanisms would appear to be involved in the VTA. PMID- 9852606 TI - Rhythmic properties of the hamster suprachiasmatic nucleus in vivo. AB - We recorded multiple unit neural activity [multiunit activity (MUA)] from inside and outside of the suprachiasmatic nucleus (SCN) in freely moving male golden hamsters housed in running-wheel cages under both light/dark cycles and constant darkness. The circadian period of MUA in the SCN matched the period of locomotor activity; it was approximately 24 hr in wild-type and 20 hr in homozygous tau mutant hamsters. The peak of MUA in the SCN always occurred in the middle of the day or, in constant darkness, the subjective day. There were circadian rhythms of MUA outside of the SCN in the ventrolateral thalamic nucleus, the caudate putamen, the accumbens nucleus, the medial septum, the lateral septum, the ventromedial hypothalamic nucleus, the medial preoptic region, and the stria medullaris. These rhythms were out-of-phase with the electrical rhythm in the SCN but in-phase with the rhythm of locomotor activity, peaking during the night or subjective night. In addition to circadian rhythms, there were significant ultradian rhythms present; one, with a period of approximately 80 min, was in antiphase between the SCN and other brain areas, and another, with a period of approximately 14 min, was in-phase between the SCN and other brain areas. The periods of these ultradian rhythms were not significantly different in wild-type and tau mutant hamsters. Of particular interest was the unique phase relationship between the MUA of the bed nucleus of the stria terminalis (BNST) and the SCN; in these two areas both circadian and ultradian components were always in-phase. This suggests that the BNST is strongly coupled to the SCN and may be one of its major output pathways. In addition to circadian and ultradian rhythms of MUA, neural activity both within and outside the SCN was acutely affected by locomotor activity. Whenever a hamster ran on its wheel, MUA in the SCN and the BNST was suppressed, and MUA in other areas was enhanced. PMID- 9852607 TI - Control of grip force when tilting objects: effect of curvature of grasped surfaces and applied tangential torque. AB - When we manipulate objects in everyday tasks, there are variations in the shape of the grasped surfaces, and the loads that potentially destabilize the grasp include time-varying linear forces and torques tangential to the grasped surfaces. Previous studies of the control of fingertip forces for grasp stability have dealt principally with flat grip surfaces and linear force loads. Here, we studied the regulation of grip force with changes in curvature of grasped surfaces and changes in tangential torque applied by the index finger and thumb when humans lifted an object and rotated it about the horizontal grip axis through an angle of 65 degrees. The curvatures of the matched pair of spherical surfaces varied from -50 m-1 (concave with radius 20 mm) to 200 m-1 (convex with radius 5 mm). The applied tangential torque at the orientation of 65 degrees was varied sixfold. Regardless of the values of curvature and end torque, grip force and tangential torque were coordinated, increasing in parallel throughout the tilt with an approximately linear relationship; the slope of the line increased progressively with increasing surface curvature. This parametric scaling of grip force was directly related to the minimum grip force required to prevent rotational slip, resulting in an adequate safety margin against slip in all cases. We conclude that surface curvature parametrically influences grip force regulation when the digits are exposed to torsional loads. Furthermore, the sensorimotor programs that control the grip force apparently predict the effect of the total load comprising linear forces and tangential torques. PMID- 9852608 TI - Perinatal gonadectomy exerts regionally selective, lateralized effects on the density of axons immunoreactive for tyrosine hydroxylase in the cerebral cortex of adult male rats. AB - The catecholamine innervation of the cerebral cortex is essential for its normal operations and is implicated in cortical dysfunction in mental illness. Previous studies in rats have shown that the maturational tempo of these afferents is highly responsive to changes in gonadal hormones. The present findings show that perinatal hormone manipulation also has striking, region- and hemisphere-specific consequences for cortical catecholamines in adulthood. The effect of perinatal gonadectomy on catecholamines was examined in representative sensory, motor, and association cortices of adult male rats by combining hormone manipulation with immunocytochemistry for tyrosine hydroxylase, a rate-limiting enzyme in catecholamine biosynthesis. Qualitative and quantitative comparison of immunoreactivity in rats perinatally gonadectomized or sham-operated revealed complex changes in gonadectomized subjects; in cingulate cortex, TH immunoreactivity was strongly and bilaterally diminished, in sensory and motor cortices, axon density was decreased in left hemispheres, but was minimally affected on the right, and in a premotor cortex, gonadectomy was without significant effect in either hemisphere. Corresponding analyses in gonadectomized rats supplemented with testosterone revealed a protective influence, albeit one in which TH immunoreactivity so showed regional and hemispheric variability in responsiveness to hormone replacement. These complex patterns of TH sensitivity suggest highly asymmetric hormone stimulation of cortical catecholamines. Such discriminative action may contribute to sex differences in the functional maturation and lateralization of the cortex and may also have bearing on disorders such as dyslexia, which show sexual dimorphisms, and in which functional laterality of the cortex may be particularly at issue. PMID- 9852610 TI - Isoprenoid synthesis gene for geranylgeranyl diphosphate synthase from a hyperthermophile, Pyrococcus sp. strain OT3. AB - Pyrococcus sp. strain OT3, a hyperthermophilic archaeon that was isolated by the authors was found to contain tetraether lipid mainly in the membrane lipid, which was quite different from the other hyperthermophiles (Masuchi et al. 1997). Those isoprenoids are synthesized by a family of isoprenyl diphosphate synthases from isopentenyl diphosphate to allylic diphosphates. The gene that encodes one of these families, geranylgeranyl diphosphate synthase (GGPPSase), from this strain was cloned and sequenced. This coding gene has a 960-bp (320aa) sequence. The putative Shine-Dalgarno sequence was six bases upper of start codon, exactly the same as Methanobacterium thermoautotrophicum, a methnogenic thermophile. Comparison of the amino acid sequence of 13 organisms including Eukarya, Bacteria, and Archaea showed that Archaea strains including Pyrococcus sp. strain OT3 consisted of a separate group from the others, but five conservative regions are very homologous. PMID- 9852609 TI - Glutamate inhibits GABA excitatory activity in developing neurons. AB - In contrast to the mature brain, in which GABA is the major inhibitory neurotransmitter, in the developing brain GABA can be excitatory, leading to depolarization, increased cytoplasmic calcium, and action potentials. We find in developing hypothalamic neurons that glutamate can inhibit the excitatory actions of GABA, as revealed with fura-2 digital imaging and whole-cell recording in cultures and brain slices. Several mechanisms for the inhibitory role of glutamate were identified. Glutamate reduced the amplitude of the cytoplasmic calcium rise evoked by GABA, in part by activation of group II metabotropic glutamate receptors (mGluRs). Presynaptically, activation of the group III mGluRs caused a striking inhibition of GABA release in early stages of synapse formation. Similar inhibitory actions of the group III mGluR agonist L-AP4 on depolarizing GABA activity were found in developing hypothalamic, cortical, and spinal cord neurons in vitro, suggesting this may be a widespread mechanism of inhibition in neurons throughout the developing brain. Antagonists of group III mGluRs increased GABA activity, suggesting an ongoing spontaneous glutamate mediated inhibition of excitatory GABA actions in developing neurons. Northern blots revealed that many mGluRs were expressed early in brain development, including times of synaptogenesis. Together these data suggest that in developing neurons glutamate can inhibit the excitatory actions of GABA at both presynaptic and postsynaptic sites, and this may be one set of mechanisms whereby the actions of two excitatory transmitters, GABA and glutamate, do not lead to runaway excitation in the developing brain. In addition to its independent excitatory role that has been the subject of much attention, our data suggest that glutamate may also play an inhibitory role in modulating the calcium-elevating actions of GABA that may affect neuronal migration, synapse formation, neurite outgrowth, and growth cone guidance during early brain development. PMID- 9852611 TI - An active Ac-like transposable element in teleost fish. AB - The i4/i4 genotype of the medakafish, Oryzias latipes, exhibits a quasi albino phenotype due to insertion of a novel transposable element, Tol2, into the tyrosinase gene. Tol2 is 4681 bp in length, has short inverted terminal repeats, and contains four ORFs with the potential to encode a transposase protein. Excision activity of the element has been detected by PCR analysis. Genomic Southern of the Tol2 element revealed that about 20 copies are present in the diploid genome. Dot-matrix comparisons of amino acid sequences of ORFs show relatively high similarity with transposases from Ac of maize, hobo of Drosophila, and Tam3 of snapdragon, which are all active transposable elements. Tol2 is thus concluded to be an active Ac-like transposable element probably encoding a transposase protein. It should therefore find application as a unique material for establishing a gene tagging system in fish. PMID- 9852612 TI - Antitumor glycogen from scallops and the interrelationship of structure and antitumor activity. AB - Hot water extract of scallop was treated with actinase E and fractionated by Sephadex G-25 gel-filtration and DEAE Sephadex A-25 ion-exchange chromatography. The antitumor activity of these fractions against Meth-A fibrosarcoma was examined. The nonadsorbed fraction (SCA25A) and weakly adsorbed fraction (SCA25B) obtained on DEAE Sephadex A-25 anion-exchange gel showed strong antitumor activity. Chemical analyses and NMR spectra identified SCA25A and SCA25B as glycogen. However, glycogen extracted from the scallop with trichloroacetic acid and from abalone showed no antitumor activity. This difference was thought to be due to variations in the fine structure of the glycogen molecule. The fine structure of glycogen was investigated by a sequential enzyme digestion method using beta-amylase and pullulanase, while the unit chain was analyzed by high performance anion exchange chromatography. The results showed that the antitumor active glycogen was highly branched with a shorter chain than glycogens without antitumor activity. PMID- 9852613 TI - Isolation of cDNAs for trypsinogen from the winter flounder, Pleuronectes americanus. AB - Knowledge of the timing of digestive enzyme expression in developing larvae is essential for evaluating the appropriateness of formulated larval diets. Since little is known at the molecular biological level about the ontogeny of digestive enzyme function in flatfish, we have isolated cDNA clones for key digestive enzymes such as trypsinogen. Portions of trypsinogen genes have been amplified from winter flounder cDNA libraries by PCR using primers based on sequence motifs conserved among trypsinogen genes from other organisms. The PCR products were sequenced, cloned, and used as radioactively labeled probes to screen the libraries. Three distinct trypsinogen cDNAs have been isolated, representing the first cDNAs for winter flounder digestive enzymes. The first type (Flounder 1) is very similar to a trypsinogen sequence reported from a related flounder, Pleuronectes platessa. The second type (Flounder 2) is related to sequences reported from Atlantic salmon, cod, and the Antarctic fish, Paranotothenia magellanica. The third type (Flounder 3) shows limited similarity to the Antarctic fish trypsinogen sequence. All three cDNAs encode a cleavable signal sequence at the amino-terminal end, and Flounder 1 and 2 contain the characteristic acidic sequence for activation of the trypsinogen proenzyme to trypsin. Southern hybridization experiments indicate that Flounder 1 and 3 are single-copy genes, whereas Flounder 2 may be present in more than one copy. In addition, Flounder 2 detects homologs in a wide variety of other fish species. The sequence information will be used to establish RT-PCR assays for larval winter flounder (Pleuronectes americanus) digestive enzyme expression. PMID- 9852614 TI - Cryopreservation of Eisenia bicyclis (Laminariales, Phaeophyta) in liquid nitrogen. AB - Gametophytic cells of Eisenia bicyclis (Kjellman) Setchell were cryopreserved in liquid nitrogen (LN). Ethylene glycol was the most effective cryoprotectant when used alone, and in combination with 10% (w/v) proline improved the survival. Viability of female and male cryopreserved cells after thawing reached 62.0% and 52.6% immediately after thawing, but these levels decreased to 31.1% and 27.2% after 4 days postthawing culture. The optimal prefreezing temperature was -40 degreesC, and cells prefrozen to temperatures >-40 degreesC were damaged mainly by intracellular ice crystal formation during immersion in LN, while those prefrozen to temperatures <-40 degreesC were damaged mainly by excessive dehydration during prefreezing. The survival rates were not affected after storage in LN for at least 200 days. PMID- 9852615 TI - Nutrient regeneration in coastal seas by Noctiluca scintillans, a red tide causing dinoflagellate. AB - Ammonium nitrogen, phosphate, and silicate contents in Noctiluca scintillans cell fluid were estimated at 2470, 183, and 54 pmol per individual, respectively. Ammonium nitrogen and phosphate concentrations at the surface layer of the water column seemed to be related to cell abundance of N. scintillans at the sampling location. Patches of N. scintillans provided 16 and 25 times greater concentrations of ammonium nitrogen and phosphate in the uppermost layer (0-10 cm depth) of the water column than those in the ambient seawater, respectively. Silicate concentration within patches, however, was close to that in surrounding water. The morphology of fecal pellets of N. scintillans fed on the diatom, Thalassiosira sp., was examined with a scanning electron microscope. The fecal pellet was composed entirely of visible structural diatom cells. It could be inferred that N. scintillans excretes silica from the diatom undigested in its fecal pellets and thus no remarkably high silicate concentration within red tide patches was observed. PMID- 9852616 TI - Antifouling activity of Indian marine invertebrates against the green mussel Perna viridis L. AB - Thirty-one species of marine invertebrates collected from the Indian Coast, belonging to six phyla (Porifera, Coelenterata, Annelida, Mollusca, Echinodermata, and Eurochordata) were tested for antifouling activity against the green mussel Perna viridis L. The repulsive action of the foot and production of byssus threads by green mussels were used for assaying the above activity. Subergorgia suberosa, Sinularia numerosa, Sinularia compressa, Cladiella pachyclados (phylum Coelenterata), Haliclona sp. (phylum Porifera), and Planaxis sulcatus (phylum Mollusca) have shown to be potential sources of antifoulants. PMID- 9852617 TI - Metabolites from the sponge-associated bacterium Micrococcus luteus. AB - In an ongoing survey of the bioactive potential of microorganisms associated with marine invertebrates, the extract of the sponge-associated bacterial strain Micrococcus luteus was found to exhibit potent antimicrobial activity. The previously known synthetic 2,4,4'-trichloro-2'-hydroxydiphenylether was found to be responsible for the antimicrobial activity. The major metabolite isolated was a new acyl-1-(acyl-6'-mannobiosyl)-3-glycerol. PMID- 9852618 TI - Continuous and simultaneous cultivation of benthic food diatom Nitzschia sp. and abalone Haliotis sieboldii by using deep seawater. AB - By using low-temperature, clean, and nutrient-rich properties of deep seawater (DSW; seawater below the euphotic layer), a continuous and simultaneous cultivation system for a benthic food diatom, Nitzschia sp., and juvenile abalone was established. Cell suspension of Nitzschia sp. was added to a bioreactor made of acrylic pipe (7 cm diameter x 50 cm long) containing short vinyl tubes (2 cm diameter x 2 cm long) as substrata. DSW collected from 320 m depth at Muroto City, in Kochi Prefecture, Japan, was supplied to the reactor and incubated under natural light (ca. 6000 lux) with a continuous DSW flow rate of 40 turnovers per hour. After growing enough benthic diatoms in the reactor, juveniles of abalone, Haliotis sieboldii (shell length ca. 10-20 mm) were put into a reactor, and cultivated simultaneously with food diatoms in the continuous flow system. During the four-month incubation, 7-month-old abalone juveniles with a shell length of 12.4 (average) +/-0.2 (SD) mm were grown to 19.4 (+/-1.7) mm in the reactor. Daily growth rates of abalones were 50-110 um/day. These results indicate that the continuous cultivation system with DSW supports the growth of juvenile abalone without any supply of seaweed until it grows to release size. PMID- 9852619 TI - Induction of apoptosis by a marine microalgal polysaccharide in a human leukemic cell line. AB - An extracellular polysaccharide produced by a marine microalga Gymnodinium A3 OKU 1 strain showed a strong cytotoxicity for the human leukemia K-562 cells (CC50 = 11.6 ug/ml). We revealed that this polysaccharide induced apoptosis of K-562 cells, as shown by cell morphology, nuclear fragmentation, in situ end-labeling of fragmented DNA, and internucleosomal DNA fragmentation, suggesting that the mechanism of cytotoxicity of this polymer was based on the induction of apoptosis. PMID- 9852620 TI - Antioxidative activity for human low density lipoprotein oxidation by a novel compound purified from marine microbial origin. AB - This study was designed to investigate the antioxidative activity of a substance isolated and purified from culture broth of Bacillus sp. The antioxidative substance was identified as 2,6-bis(1,1-dimethylethyl)phenol (Phen-251). The antioxidative activity of Phen-251 was higher than that of dl-tocopherol and BHA against low-density lipoprotein (LDL) oxidation by thiobarbituric acid reactive substance (TBARS). Phen-251 inhibited the copper-mediated oxidation of human LDL in a dose-dependent manner with almost complete inhibition at 40 ug/ml. Phen-251 at a concentration of 60 ug/ml also inhibited oxidation of LDL induced by J774. LDL oxidized by copper-mediated or cell-induced oxidation was degraded at a much greater rate than native LDL. These results suggested the importance of further research on Phen-251 in the investigation of atherosclerosis and free radical induced injury. PMID- 9852621 TI - Genetic structure in wild populations of black tiger shrimp (Penaeus monodon) using randomly amplified polymorphic DNA analysis. AB - Randomly amplified polymorphic DNA (RAPD) analysis was used to examine genetic variation in wild black tiger shrimp, Penaeus monodon. Specimens were collected from five geographically separated locations (Satun-Trang, Phangnga, and Medan in the Andaman Sea and Chumphon and Trad in the Gulf of Thailand). A total of 100 P. monodon individuals were investigated using seven arbitrarily selected primers. Fifty-eight (72.5%) of eighty reproducible RAPD fragments ranging in size from 200 to 2200 bp were polymorphic. The percentages of polymorphic bands of the five geographic populations investigated varied from 51.5 to 57.7%. The genetic distance between populations and UPGMA dendrograms indicated that the Medan population was genetically different from Thai P. monodon (Dij = 14.976%). Within Thailand, the Satun-Trang P. monodon was separated from the remaining geographic populations with a genetic distance of 2.632%. RAPD analysis in the present study yielded a total of 252 genotypes. A Monte Carlo analysis illustrated geographic heterogeneity in genotype frequencies within this species, suggesting that genetic population structure does exist in this taxon (P < 0.001 for all primers). Signficant differences in genotype frequencies between Thai and Indonesian (Medan) P. monodon were observed (P < 0.0001). Within Thailand, the Andaman Sea P. monodon was significantly different from that of the Gulf of Thailand (P values between 0.0000 and 0.0387), indicating population differentiation between P. monodon from these two main fishery regions of Thailand. PMID- 9852622 TI - Cell yield and superoxide dismutase activity of the marine yeast Debaryomyces hansenii under different culture conditions. AB - The effect of aeration, pH, stirring rate, and temperature on the biomass production and superoxide dismutase (SOD) activity of the marine yeast Debaryomyces hansenii strain C-11 was determined. The cell biomass yield was approximately 50% in a seawater-formulated medium using glucose as the carbon source. The SOD activity increased by application of a pulse of oxygen or 0.8 mM sulfate copper into the chemical reactor. The SOD enzyme had an activity of 400 units/mg of protein in a crude extract produced under such conditions, the best activity ever reported for this enzyme in a crude preparation. PMID- 9852623 TI - Purification and characterization of agarases from a marine bacterium, Vibrio sp. PO-303. AB - A marine bacterium, Vibrio sp. PO-303, produced three kinds of extracellular agarases. These enzymes were purified to homogeneity by ammonium sulfate precipitation and successive column chromatographies. The molecular masses of agarase-a, -b, and -c were estimated to be 87.5, 115, and 57 kDa by SDS-PAGE with isoelectric point of 6.6, 3.4, and 8.4, respectively. These enzymes had maximal activity at pH 6.5-7.5 and at around 38-55 degreesC. They differed in their sequences at the amino termini of the protein chains. All enzymes were inhibited completely by Hg2+. Ag+, Cu2+, and Zn2+ strongly inhibited agarase-a and -c compared with agarase-b, and the activity of agarase-c fell wide by Al3+, Fe3+, and EDTA. Agarase-a hydrolyzed agarose to give neoagarotetraose and -hexaose as predominant products, but could not cleave neoagarotetraose. The main hydrolysis products of agarase-b were neoagarobiose from agarose and neoagarooligosaccharides more than dimer. Agarase-c could not cleave neoagarohexaose. PMID- 9852624 TI - Vibrio spp., the dominant flora in shrimp hatchery against some fish pathogenic viruses. AB - Two strains of Vibrio spp., NICA 1030 and NICA 1031, which are the dominant flora in shrimp hatcheries, were studied for antiviral activity by exposing them to the infectious hematopoietic necrosis virus (IHNV) and Oncorhynchus masou virus (OMV) before being inoculated to the CHSE-214 cells. Antiviral activities were exhibited as plaque reduction percentages of 98 and 92% for the NICA 1030 and of 99 and 62% for the NICA 1031, respectively. PMID- 9852625 TI - p53 and protein kinase C independent induction of growth arrest and apoptosis by bryostatin 1 in a highly metastatic mammary epithelial cell line: In vitro versus in vivo activity. AB - We have evaluated the effects of bryostatin 1 on growth of a highly malignant p53 null mouse mammary tumor line, 4T1, and the mechanism by which bryostatin 1 inhibits in vitro growth and in vivo development of tumor and metastases from the orthotopic site. Bryostatin 1 at 20-400 nM concentrations inhibits growth of 4T1 cells by approximately 60% in two-day cultures. Inhibition of growth is associated with an increase in the number of cells undergoing apoptosis with concomitant elevation in the steady state levels of bax protein and drop in bcl-2 levels. The cytotoxic effect of bryostatin 1 on 4T1 cells occurs independently of p53, since there was no evidence of p53-mediated transcriptional activity in 4T1 cells following treatment with bryostatin 1.4T1 cells respond in vivo to bryostatin 1 therapy (75 microg/kg body weight). Intraperitoneal administration of bryostatin 1 inhibits both primary and secondary tumor growth by approximately 50%. However, although bryostatin 1 has a remarkable capacity to slow tumor growth and progression, it is unable to completely eradicate tumor growth and progression due to in vivo development of tumor resistance to bryostatin 1. Levels and cellular distribution of PKCalpha and delta do not correlate with the growth inhibitory effects of bryostatin 1 on 4T1 cells; however, reduction in cytosolic PKCalpha and delta without associated increase in membrane compartment appear to correlate with bryostatin-resistance. Our results suggest that the therapeutic effects of bryostatin 1 in our system do not involve alterations in levels and distribution of PKC but rather a direct upregulation of bax/ bcl-2 ratios that is independent of p53. PMID- 9852626 TI - A heterozygous frameshift mutation of the PTEN/MMAC1 gene in a patient with Lhermitte-Duclos disease - only the mutated allele was expressed in the cerebellar tumor. AB - Lhermitte-Duclos disease is a rare disorder of the cerebellum which exhibits a focally indolent growth of the cerebellar cortex. The disease is sometimes associated with multiple hamartoma syndrome, or Cowden disease, an autosomal, dominantly inherited cancer syndrome. Germline mutations of the PTEN/MMAC1 gene, a gene recently discovered to be a tumor suppressor, have been documented in four families with Cowden disease. A family member in one of these families have had Lhermitte-Duclos disease, indicating that mutations of the gene are responsible for the development of Lhermitte-Duclos disease. However, the occurrence of Lhermitte-Duclos disease is mostly sporadic. It is unknown whether the PTEN/MMAC1 gene is involved in these cases. We performed a genetic analysis on a patient with Lhermitte-Duclos disease, as well as analyses on his family members, and found a germline mutation of the PTEN/MMAC1 gene. An insertion of A at nucleotide 83 in codon 28 was apparent in both the patient and members of his family. This mutation caused a frame shift that generated a premature stop codon in codon 43. The mutation was heterozygous, although only the mutated allele was expressed in the cerebellar tumor of the patient. A monoallelic expression of the mutated PTEN/MMAC1 gene may have been responsible for the development of a cerebellar tumor in the patient. PMID- 9852627 TI - Involvement of apoptosis and cyclin D1 gene repression in growth inhibition of T 47D human breast cancer cells by methylglyoxal bis(cyclopentylamidinohydrazone). AB - Polyamines are considered to be important intracellular molecules for the proliferation of the cancer cells. In this study, effects of methylglyoxal bis(cyclopentylamidinohydrazone) (MGBCP), a potent inhibitor of the polyamine biosynthetic pathway, on the growth and cell cycle of T-47D human breast cancer cells were investigated. MGBCP dose-dependently inhibited the growth of T-47D cells, in which the contents of spermine, spermidine and putrescine decreased concomitantly. The gene expression of cyclin D1 was also repressed by the MGBCP treatment. The MGBCP-treated cells clearly exhibited morphological changes indicating the blebbing and chromatin condensation which are characteristic of apoptosis. Flow cytometric analysis showed hypo-diploid subpopulations due to apoptotic cells, and characteristic oligonucleosomal-sized DNA fragments were clearly observed for MGBCP-treated cells as the concentration of the drug was increased. These findings suggest that the inhibition of polyamine synthesis results in the repressions of cyclin D1 expression and cell cycle progression, eventually inducing apoptosis in these human breast cancer cells. PMID- 9852628 TI - Germinated barley foodstuff increases fecal volume and butyrate production in humans. AB - Germinated barley foodstuff (GBF), derived from the aleurone layer, scutellum and germ of germinated barley, contains a large quantity of fermentable dietary fibers, especially hemicellulose. Ten grams of GBF were given to 10 healthy volunteers 3 times a day (30 g/day/person) for 28 consecutive days. Fecal weight, water contents and short chain fatty acid content were measured before GBF administration and from days 25 to 28 after initiation of GBF administration. GBF intake significantly increased fecal butyrate content as well as fecal weight and water content. No significant change in body weight resulted from consumption of GBF for 28 days. No major laboratory abnormalities were found in hematologic and urinary analysis. These findings indicate that GBF promotes defecation, produces bacterial short chain fatty acids, especially butyrate, without adverse effects, and is a safe foodstuff for humans. PMID- 9852630 TI - Molecular correlates of bcl-2-enhanced growth following androgen-ablation in prostate carcinoma cells in vivo. AB - Androgen-independent growth of prostate cancer is correlated with expression of bcl-2. The impact of bcl-2 expression on the growth of prostate cancer cells following androgen ablation, was examined in the androgen-sensitive prostatic carcinoma cell line, LNCaP. Vector control and bcl-2 expressing LNCaP cells were grown subcutaneously in male nude mice. Tumor volume, apoptosis, and proliferation were assessed following castration. The levels of c-myc, p53, p21, bax, and bcl-2 protein were assessed by Western blotting. Bcl-2 expressing tumors exhibited a significant augmentation in growth compared to controls (p 0.01). No difference in the spontaneous rate of proliferation was observed between bcl-2 and control tumors, however, bcl-2 expressing tumors exhibited lower rates of apoptosis. Following orchiectomy the apoptotic index remained significantly lower in bcl-2 expressing tumors (p 0.002 at day 3). The proliferative index was maintained in bcl-2 expressing, but not control tumors following castration. This resulted in a significant growth advantage in bcl-2 tumors subsequent to androgen ablation (p 0.001). These changes were accompanied by alterations in the levels of gene products known to regulate the cell cycle and/or apoptosis. These results emphasize the significance of bcl-2 expression during prostate cancer progression and suggest possible mechanisms for the acquisition of androgen-independent tumor growth. PMID- 9852629 TI - In vitro effects of GM-CSF on mature peripheral blood neutrophils. AB - GM-CSF can play a crucial role in regulating the neutrophil-mediated inflammatory response. This growth factor is a proliferative stimulus for bone marrow neutrophil stem cell precursors and has at least 3 important roles in regulating neutrophil-mediated immunity: a) a direct effect on the proliferation and development of neutrophil progenitors; b) synergistic activity with other haemopoietic growth factors; c) stimulation of the functional activity of mature neutrophils. The production of GM-CSF may be triggered directly by exogenous factors such as antigens and endotoxins, or indirectly through the release of cytokines by a variety of cells including lymphocytes, activated macrophages and endothelial cells exposed to products of mononuclear phagocytes. Such production of GM-CSF may serve to quickly release mature neutrophils from the bone marrow in response to infections. Moreover, enhancement of the function of mature neutrophils may also augment their ability to migrate to infective sites and then phagocytose and kill pathogens. Increased expression of CD11b/CD18 may play a fundamental part in this mechanism because this receptor is essential for the adhesion of neutrophils to the endothelium. Both phagocytosis and oxidative burst activity increase as a result of the action of GM-CSF and the increased expression of complement- and Fc-receptors can augment opsono-phagocytosis. A further level of neutrophil up-regulation occurs by increasing the functional life span of neutrophils by GM-CSF. Thus, by delaying neutrophil apoptosis, GM CSF greatly extends the time over which neutrophils may function at inflammatory sites. GM-CSF can thus exert a variety of important regulatory controls of neutrophil function during bacterial infections. Both the number and the functional status of neutrophils is highly regulated by GM-CSF. It is also possible that GM-CSF produced within localised sites of acute inflammation or infection may attract, trap and then activate neutrophils within this site. PMID- 9852631 TI - Preventive effects of germinated barley foodstuff on methotrexate-induced enteritis in rats. AB - The preventive effects of the dietary germinated barley foodstuff (GBF), which increases the contents of protein, RNA and DNA in the intestinal mucosa of rats on the mucosal damage and diarrhea were examined in a methotrexate (MTX)-induced enteritis model in rats. Sprague-Dawley rats intraperitoneally injected with MTX (10 mg/kg body weight) were used as an enteritis model. After consumption of diets containing GBF, glutamine or a glutamine-rich stuff (gluten), mucosal damage, contents of mucosal protein, RNA and DNA, myeloperoxidase (MPO) activity, bacterial translocation and DNA synthetic activity in the small intestine were assessed. GBF more effectively prevented diarrhea and mucosal damages, and increased mucosal protein, DNA and RNA contents than glutamine or gluten. The bacterial trans-location and elevation of MPO activity induced by MTX were depressed only by the consumption of GBF. GBF has a potential as therapeutic diet to decrease the adverse effects of anti-cancer chemotherapy. PMID- 9852632 TI - Effects of D-mannoheptulose and its hexaacetate ester upon D-glucose metabolism in rat hepatocytes. AB - D-mannoheptulose, which inhibits hexokinase isoenzymes in a predominantly competitive manner, has been found to decrease much more modestly D-glucose metabolism in pancreatic islets exposed to a low, as distinct from high, concentration of the hexose. In the present study, which aimed at investigating the factor(s) possibly responsible for such a phenomenon, a comparable situation was found to prevail in rat hepatocytes. However, when the hexaacetate ester of D mannoheptulose was used instead of the unesterified heptose, the relative extent of inhibition of D-[5-3H]glucose utilization and D-[U-14C]glucose conversion to 14C-labelled acidic metabolites was comparable in hepatocytes exposed to either 1.7 or 8.3 mM D-glucose. Moreover, at the low D-glucose level, the incorporation of 3-O-methyl-D-glucose (6.6 mM) into the incubation medium increased the inhibitory action of unesterified D-mannoheptulose upon D-glucose metabolism. These findings suggest that an insufficient uptake of the heptose accounts, in part at least, for its poor efficiency as inhibitor of D-glucose catabolism in liver, and presumably islet cells exposed to low concentrations of the hexose. PMID- 9852633 TI - Protein turnover in skeletal muscle of the diabetic rat: activation of ubiquitin dependent proteolysis. AB - Induction of experimental insulin-deficiency by a single administration of streptozotocin to rats resulted in substantial changes in heart and skeletal muscle size and protein content. This was accompanied by a marked loss of total body (carcass) nitrogen and raised concentrations of circulating branched-chain amino acids. These changes were related to alterations in protein turnover in skeletal muscle. Thus, the diabetic animals showed changes in both the fractional protein rates of synthesis (decreased by 37%) and degradation (increased by 141%). The increased protein degradation observed in the muscle of the diabetic animals was associated only with an increase in the expression of the genes controlling ubiquitin-dependent proteolysis. It may be suggested that the hormonal changes associated with the diabetic state play an important role in the regulation of the activity of the ubiquitin-dependent proteolytic system in skeletal muscle, highlighting the major role of this system in the diabetes related cachexia. PMID- 9852634 TI - Immunological profile of endemic and epidemic Kaposi's sarcoma patients in Dar-es Salaam, Tanzania. AB - Kaposi's sarcoma (KS) presents in four clinicopathological types namely classical/sporadic (CKS), endemic African (EKS), iatrogenic (IKS) and that associated with AIDS (AKS). Recently a putative herpes virus (HHV-8) was described and shown to be present in all four types of KS. The immunological status of patients with EKS has been conflicting. In this study total leucocyte counts, total lymphocyte counts and lymphocyte subsets of patients with EKS and AKS were determined by flow cytometry and compared to those of healthy HIV-1 seronegative controls. Results show that 50% of EKS lesions were of nodular type. Patients with EKS had significantly lower levels of CD4+ T- lymphocytes and CD4:CD8 ratio but significantly higher CD8+ T-lymphocytes compared to controls. Patients with AKS had significantly lower levels of CD4+ T-lymphocytes and also CD4:CD8 ratios but significantly higher percentage of CD8+ T-lymphocytes when compared with EKS patients. These findings indicate that in both forms of KS there is a certain degree of immunological disturbance which is more conspicuous in AKS because of HIV infection and suggests that HIV-1 acts synergistically with the aetiological agent (HHV-8) to cause a more aggressive type of KS. PMID- 9852635 TI - Molecular analysis of critical sequences within the EBNA-2 type 1 gene from Epstein-Barr virus isolates from patients with infectious mononucleosis, tonsillar hyperplasia, and HIV infection. AB - EBNA-2 is the first protein to be detected after infection of primary B lymphocytes by Epstein-Barr virus (EBV) and plays an essential role as transcriptional activator in EBV-induced lymphocyte transformation. We analysed by PCR and sequencing regions of the EBNA-2 type 1 gene from isolates from 13 children with infectious mononucleosis (IM), 6 children with tonsillar hyperplasia (TH), and 9 patients with HIV infection followed longitudinally. We found in all three groups of patients frequent non-silent point mutations at positions 48990, 48991, 49021, 49057, 49083, 49089, 49091, 49113, 49119, 49140, 49156, and a triplet insertion at position 49136. While 4 out of 13 samples from patients with IM showed a mosaic pattern suggesting co-existence of more than 1 substrain of EBNA-2 type 1, none of the samples from TH showed this pattern consistent with substrain selection during clinical latency. No sequence changes were noted over time in samples derived from patients with HIV infection. We conclude that in analogy to the coexistence of several subtypes of EBNA-1 in healthy EBV carriers, samples from IM can harbor more than one subtype of the EBNA-2 type 1 gene. PMID- 9852636 TI - Detection of polymorphic triplet repeats in the genomes of patients suffering from bipolar affective disorder. AB - Evidence for the operation of expanded trinucleotide repeats in the pathogenesis of bipolar affected disorder has recently been found at the molecular genetic level. For the screening of these repeat motifs in genomes of patients with bipolar affective disorder, we established a modified PCR-based fingerprinting technique, called triplet repeat enhanced arbitrarily primed PCR (TREAP-PCR). Using this approach, 40 patients suffering from bipolar affective disorder (ICD10: F31) and 15 healthy controls were investigated. Interindividual polymorphisms generated by TREAP-PCR seemed to depend on the type of triplet. Using CCG triplet primers, polymorphisms could be observed more often in the genomes of patients compared with controls, whereas no significant differences could be found using primers of the CAG or AAT type. These data might indicate the existence of subgroups of manic-depressive patients based on molecular genetic differences. PMID- 9852637 TI - Detection of cytokine gene expression in human monocytes and lymphocytes by fluorescent in situ hybridization in cell suspension and flow cytometry. AB - The use of digoxigenin (DIG)- and biotin-labelled dsDNA probes to detect TNFalpha mRNA accumulation in human peripheral blood mononuclear cells (PBMC) and isolated monocytes is described. The fragment of the glyceraldehyde-3-phosphate dehydrogenase GAPDH-cDNA was used as a control probe. The hybridization signals were detected by staining with fluorescein isothiocyanate (FITC)-labelled anti DIG antibody and avidin-FITC, respectively. The cells were stimulated in vitro with lipopolysaccharide (LPS) for 0.5-6 h. The TNFalpha-mRNA was detected in monocytes 1 h after stimulation with LPS, and the highest accumulation was seen around 2 h. The TNFalpha-mRNA in stimulated PBMC was detected at the lower level peaking around 4 h. The TNFalpha-mRNA accumulation was lower in lymphocytes than in monocytes when PBMC were studied. There was no difference in the level of GAPDH-mRNA between unstimulated and stimulated cells. Finally, an enhanced accumulation of TNFalpha-mRNA was observed in PBMC from some patients with sepsis or cancer. Thus, this study shows that cytokine gene expression may be detected in cells ex vivo. This opens the possibility of studying the level of cytokine gene activation in PBMC of patients with diseases where the role of cytokines in their pathophysiology is implicated. PMID- 9852638 TI - The genotoxicity and cytotoxicity of dermally-administered cadmium: effects of dermal cadmium administration. AB - Cadmium, unlike zinc, selenium and copper, has no known biological importance, and therefore, it is classified as a carcinogen in humans, as well as in animals. The effect(s) of levels of dermally-administered cadmium on cadmium genotoxicity and cytotoxicity was investigated in Harlan Sprague-Dawley rats for 14, 21, 28, 35 and 42 days at concentrations of 14 and 28 mg/kg/day. Exposure of rats to cadmium via dermal application caused lesions on the skin (hyperkeratosis, acanthosis and scabbing, alopecia and erythema) and tumors in the scrotum. Anatomical changes, such as distention of the stomach, atrophy of kidney and liver and loss of body weight were also observed in these rats. The toxic effects of cadmium on cell ultrastructure were nuclear membrane damage, chromatin condensation, regression of mitochondrial cristae and ultimately cell death. Analyses of the brain, kidney and liver cells of rats exposed to cadmium, clearly showed DNA damage. Of the three organs examined, DNA from kidney cells sustained the most damage followed by DNA in liver cells. There is a positive correlation between Cd dose(s) and duration of exposure and the extent of DNA damage. PMID- 9852639 TI - Identification of endothelial antigens relevant to transplant coronary artery disease from a human endothelial cell cDNA expression library. AB - Accelerated transplant coronary artery disease (TxCAD) results in increased expression of antiendothelial antibodies whose target antigens remain largely unidentified. One of these endothelial antigens has been identified as vimentin, a cytoskeletal protein present in cells of the blood vessel walls. In the present study, SDS-PAGE and Western blot analysis of human endothelial cell (EAHy 926) lysates probed with sera from a TxCAD patient were used to confirm immunoreactivity of antiendothelial antibodies towards several endothelial proteins. To further elucidate the identity of these putative antigens, a human endothelial cell (EAHy 926) cDNA expression library was immunoscreened with serum obtained from a TxCAD patient. Two positive cDNA clones were identified by partial nucleotide sequence analysis and GenBank/EMBL database searches for homology as the 85 kDa human CD36 antigen (a cell surface glycoprotein expressed in various cells including epithelial and endothelial cells) and a 50 kDa keratin like protein (a member of the intermediate filament protein expressed in epithelial cells). These results are the first to demonstrate that human CD36 antigen and a keratin-like protein may be additional target proteins for the anti endothelial antibodies associated with TxCAD. PMID- 9852640 TI - Molecular approach in human tumor investigation: oncogenes, tumor suppressor genes and DNA tumor polyomaviruses (review). AB - Molecular analysis are useful for diagnosis, prognosis and follow-up of the patients, as well as for addressing therapeutic choices. Most of the molecular methods are based on the analysis of nucleic acids. The DNA and RNA methodologies of routine applicability include Southern and Northern hybridizations and polymerase chain reaction (PCR) techniques. Southern blot hybridization recognizes major DNA rearrangements, and detection of oncogenic viral sequences present in high copy number, whereas PCR-based methods allow the detection of gross chromosomal modifications, fine gene alterations and low amount of tumor virus footprints. PCR techniques also allow the analysis of the partially degraded nucleic acids from formalin-fixed paraffin-embedded tissues. We present an overview of the use of molecular techniques for the analysis, diagnosis, prognosis and follow-up of neoplastic diseases, using examples from our experience in both leukemias and solid tumors. PMID- 9852641 TI - Post graduate education. PMID- 9852642 TI - [Alcohol in clinical medicine]. PMID- 9852644 TI - [Risk factors related to esophageal cancer in Rio Grande do Sul, Brazil]. AB - BACKGROUND: Rio Grande do Sul, a State in southern Brazil, shows high mortality rates for esophageal cancer. Important geographical variations suggest external factors in the etiology. OBJECTIVE: The relationship between smoking, alcohol and "mate" to esophageal cancer. METHOD: 55 patients with esophageal cancer were interviewed, soon after the endoscopy, about risk factors; 110 patients who did not showed esophageal tumor in the endoscopy were the controls. RESULTS: Among the patients with esophageal cancer there were more farmer-workers (OD 3.3; 95% CI 0.9-11.2), father with antecedent cancer (OD 6.9; 95% CI 1.9-25.6), smoking and ex-smoking (OD 2.5; 95% CI 1.1-5.9), drinkers and ex- of alcohol (OD 5.3; 95% CI 2.6-11.0), drinkers and ex- of "mate" (OD 3.6; 95% CI 1.3-9.8). Others variables more frequent in the cases were: smoking for > 20 years; kind of alcohol (spirits), drinking alcohol every day, drinking "mate" every day and adding salt to food. CONCLUSIONS: Smoking, alcohol, "mate", farmer-workers and antecedent of father with cancer were significantly more frequent in the cases of esophageal cancer than in the controls. PMID- 9852643 TI - [Effect of nosocomial bacteremia caused by oxacillin-resistant Staphylococcus aureus on mortality and length of hospitalization]. AB - OBJECTIVES: To identify the attributed mortality rate of bloodstream hospital infection by Staphylococcus aureus resistant to methicillin (MRSA) and its effect on length of hospital stay. DESIGN: Case-control study. SETTING: Hospital Sao Paulo da Universidade Federal de Sao Paulo, a 660-bed, tertiary-care teaching hospital in Sao Paulo, Brazil. PATIENTS: Seventy one adults patients with hospital-acquired MRSA bacteremia diagnosed between January 1, 1991, and September 30, 1992, and 71 MRSA-free controls were matched by the following criteria: age, sex, underlying disease, surgical procedure, same risk time and admission date. RESULTS: The incidence of patients with hospital sepsis by MRSA accounted for 73.22% of the patients with hospital bloodstream infection by Staphylococcus aureus. The mortality rate of the cases was 56.33 (40/71) and 11.26 (8/71) of the controls. The attributable mortality rate was 45.07% (OR = 17.0; IC 95% = 3.58-202.26; p = 0.000001). The length of hospital stay median time was of 32.55 days for the cases and 29.75 for the controls (p = 0.32). CONCLUSION: A high level of sepsis by MRSA was observed in all the Staphylococcus aureus bacteremia. The bloodstream hospital infection by MRSA itself does provide a high level of mortality independently from the patients base disease, without however, increasing their hospital length of stay. PMID- 9852645 TI - [Alpha-fetoprotein: normal values in amniotic fluid between 14 and 21 weeks]. AB - BACKGROUND: To define the normal values of amniotic fluid alphafetoprotein in pregnant women, whose gestational ages range from 14 to 21 weeks, in the Hospital de Clinicas de Porto Alegre. MATERIAL AND METHOD: One hundred thirty seven women with indication for amniocentesis were studied. The alphafetoprotein was measured in all samples using enzyme immunoassay. One hundred and nine normal pregnancies were selected. All of these fetuses had normal karyotype and had no malformation. They were not twins and their amniotic fluid samples were not bloody. These samples were divided by their gestational ages. Then the medians of the alphafetoprotein values and their multiples were calculated. RESULTS: The medians of alphafetoprotein (KUI/ml) for each gestational age were as follows: 14 weeks: 16.32; 15 weeks: 14.36; 16 weeks: 13.43; 17 weeks: 10.93; 18 weeks: 8.22; 19 weeks: 7.35; 20 weeks: 5.62; 21 weeks: 4.47. CONCLUSION: The establishment of alphafetoprotein normal values in our service allows us to use this assay for patients at risk of neural tube defects. It also makes possible to analyze samples sent for cytogenetic or metabolic studies, in order to identify elevated levels of alphafetoprotein, so that these fetuses could have a more detailed sonography study to look for malformations. PMID- 9852646 TI - [IgA deficiency: clinical and laboratory evaluation of 60 patients from the "Instituto da Crianca"]. AB - OBJECTIVES: To evaluate the main clinical findings that suggest DIgA and describe the complications observed. PATIENTS AND METHOD: Sixty IgA deficient patients (IgA < 5mg/dl) were included, retrospectively. They were submitted to clinical evaluation and specific laboratorial tests. Their evolution was followed up to 15 years. RESULTS: The main complaints were: recurrent infections (50%), allergies (34%) or autoimmune diseases (10%). The respiratory system was the most affected by infections, and asthma and rhinitis were the most frequent allergic symptoms. No immunological impairment was detected, except for IgA deficiency. IgG and IgM levels were elevated in 50% of the patients. There was improvement of clinical symptoms and some of the patients became asymptomatic during the follow-up. CONCLUSION: IgA deficiency has a large clinical spectrum and early diagnosis would indicate prophylaxis for infections and allergy. PMID- 9852647 TI - [In vitro antimicrobial activity of cefpirome compared to other broad-spectrum beta-lactam drugs against 804 clinical isolates from 9 Brazilian hospitals]. AB - OBJECTIVE: To evaluate the in vitro activity of the fourth-generation cephalosporin cefpirome in comparison to that of ceftazidime, ceftriaxone, cefotaxime and imipenem in a multicenter study involving nine hospitals from six cities (four States). MATERIAL AND METHOD: A total of 804 isolates from patients hospitalized in either intensive care units or Oncology/Hematology units was evaluated. The isolates were collected between June and November of 1995, i.e. before cefpirome became commercially available in Brazil, and susceptibility tested by broth microdilution following the NCCLS procedures. All isolates resistant to cefpirome were retested by E-test. RESULTS: Against Enterobacteriaceae (n = 344), cefpirome demonstrated an activity 2 to 32-fold higher than that of the third-generation cephalosporins (TGCs) and similar to that of imipenem. The percentages of Enterobacteriaceae susceptible were: 88%, 69% and 96% for cefpirome, TGCs and imipenem, respectively. The cefpirome spectrum was greater or equal than that of imipenem against Citrobacter freundii, Enterobacter aerogenes, Morganella morganii and Serratia marcescens. Against Acinetobacter sp. (n = 77), cefpirome was slightly more active than ceftazidime; however, the percentages of isolates resistant to these compounds were high (84% and 88%, respectively). The activities of cefpirome, ceftazidime and imipenem were very similar against P. aeruginosa isolates (n = 128), with MIC50(mg/ml)/percent susceptible of 8/59%, 8/62% and 4/62% respectively. Against aerobic gram-positive bacteria, the cefpirome activity was 4 to 16-fold higher than that of TGCs but 2 to 8-fold lower than that of imipenem. CONCLUSION: The results suggest that, in Brazil, cefpirome has a spectrum of activity which is higher than that of the TGCs against aerobic gram-negative (Enterobacteriaceae and non-Enterobacteriaceae) and gram-positive bacteria and similar to that of imipenem against some Enterobacteriaceae species and P. aeruginosa. PMID- 9852648 TI - Long-term thalidomide use in refractory cutaneous lesions of systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the efficacy of long-term thalidomide treatment in cutaneous lesions of systemic lupus erythematosus (SLE), not responsive to conventional therapy. PATIENTS AND METHODS: Were selected 18 SLE patients (ACR criteria) with active cutaneous lesions not responsive to chloroquine, photoprotectors and low doses prednisone and who presented good response to thalidomide but relapsed after withdrawal of the drug. All female patients had no risk of pregnancy. Thalidomide was reintroduced and maintained at low dose (25 100 mg/day) for a minimum of 6 months. RESULTS: Eighteen patients (16 females) with mean age of 34.2 yo (16-57 y.o.) received thalidomide for 6-21 months (mean 8.5 m). The mean dose of prednisone at beginning of study was 38.3 mg/d and at the end was 9.7 mg/d (p < 0.05). Complete remission of cutaneous lesions was observed in thirteen patients (72%) and partial remission in five (28%). Side effects observed were: drowsiness in eight patients, intestinal constipation in 5, transient oliguria in 1, paresthesia of hand with normal electromyography in another one. All side effects disappeared with reduction of thalidomide dose and no patient needed to stop treatment owing to side effect. CONCLUSION: Thalidomide is a good alternative therapy to SLE patients with refractory cutaneous lesions and without any risk of pregnancy. PMID- 9852650 TI - [Clinical reasoning--the diagnostic and therapeutic decision process]. AB - The goal of this review is to demonstrate the steps and the main items of the cognitive process used by doctors in clinical reasoning of diagnostic and therapeutic decisions. The clinical problem-solving process makes use of the hypothetic deductive scientific method to solve problems. As soon as the doctor meets his (her) patient, many diagnostic hypotheses emerge in his (her) mind, which are evaluated and refuted or corroborated. The diagnostic decision occurs when a hypothesis reach a certain degree of likelihood. The therapeutic decision is based on the intended objectives and the waited effectiveness among many available alternatives. PMID- 9852649 TI - [Evaluation of nutritional recovery pattern of malnourished children examined at the center of Recovery and Educational Nutrition, Brazil]. AB - PURPOSE: To assess the pattern of nutritional recovery in malnourished children. This prospective analysis was conducted between September 1995 and November 1996 at the Center for Recovery and Nutrition Education (CREN) in Sao Paulo, Brazil. Of the thirty-eight children studied, 47.4% were infants (nine girls and nine boys) and 52.6% preschool children (twelve boys and eight girls). METHOD: Thirty eight children were studied by velocity of W/A and H/A gain in standard deviation or Z score, according to weight and height median of NCHS. RESULTS: There was a statistically significant increase in the weight-for-age (W/A)(p = 0.012), height for-age (H/A) (p = 0.001) from the time of admission. The difference in the velocity of W/A gain between infants (0.299-Z/year) and preschool children (0.684 Z/year) with more than six months of treatment, was found to be statistically significant (p = 0.038). The difference in the velocity of H/A gain for infants (0.794-Z/year) and for preschool children (0.506-Z/year), was not statistically significant (p = 0.166). In regards to gender, there was not a statistically significant difference between boys and girls in the velocity of W/A gain (0.540 and 0.524-Z/year, respectively). The difference in velocity of H/A gain was statistically significant, 0.485-Z/year for boys and 0.856-Z/year for girls (p = 0.043). CONCLUSION: These findings show that there are differences in the nutritional recovery patterns between gender and age groups. PMID- 9852651 TI - [Deaths caused by venomous snakes in the State of Sao Paulo: evaluation of 43 cases from 1988 to 1993]. AB - The prognostic factors and the causes for obit occurrence in ophidian envenoming are yet not completely clear. PURPOSE: To determine the prognostic factors and the most probable causes for obit occurrence in ophidian envenoming. METHODS: In the State of Sao Paulo were notified 12,639 cases of accidents by venomous snakes from 1988 to 1993. There were 43 deaths (0.34%). The variables from the accident notification reports were compared with the promptuary notes and/or with the death records (in lethal cases). RESULTS: The snake genus was classified in 11,297 accidents and in 41 from the lethal ones. Bothrops was responsible for 9,828 (87%) accidents and 28 (68.3%) deaths, Crotalus for 1,359 (12.0%) accidents and 13 deaths (31.7%) and Micrurus for 110 (1%) accidents and no death (p < 0.05). The information regarding to sex and age of the patients was available, respectively, in 12,620 and 12,527 accidents and in all lethal ones. There were 9,783 (77.5%) male patients in the accidents and 35 (81.4%) in the lethal cases (p > 0.05). Regarding to age, 15.9% of the patients and 41.8% of the ones who died were 50 years-old or more (p < 0.05). The most frequently bitten anatomic regions were: foot (42.2%), hand (20.6%), leg (17.6%), and ankle (13.1%) in the accidents, and foot (35.7%) and leg (35.7%) in the lethal cases (p < 0.05). Coagulation disorders occurred in 34 (91.9%) from 37 patients who died (those where this datum was available). The information above was not available in non lethal cases. The most common manifestations and complications implicated as possible death causes were: acute renal failure (34-79.1%), acute respiratory failure (28-65.1%), shock (18-41.9%), and sepsis (18-41.9%). Among all lethal cases but one without information, 29.4% of the patients died within the first two days after bite and 67.6% within the first 5 days. Acute respiratory failure was most common among the patients who died owing to crotalic envenoming, and sepsis was only seen in bothropic envenoming. CONCLUSION: Most of the accidents and obits are caused by Bothrops; Crotalus envenoming, leg bites, and accidents in 50 year-old patients (or more) are frequently lethal; the most common complication in lethal cases is acute renal failure. PMID- 9852652 TI - [Experience of the Baby Friendly Hospital initiative]. AB - In the study is analyzed and described the initiative called "Initiative Baby Friendly Hospitals", a program which started in Brazil, 1992. This initiative intends to support, to protect and to promote the breastfeeding as proposed in a meeting in 1990 in Florence, Italy, which was promoted by WHO and UNICEF. The basic goal of this initiative is to mobilize health professionals and hospital or maternity workers for changing their routines and conducts aiming to prevent the early wean. The health establishments are evaluated based on the "ten steps for success of breastfeeding, a group of goals created in the same meeting. In Brazil, the evaluation is coordinated by the Federal Government through the PNIAM (Programa Nacional de Incentivo ao Aleitamento Materno). A baby friendly hospital, if approved, receives from the Minister of Health, a Federal Governmental Agency (SUS) a differential payment for childbirth assistance and prenatal accompaniment, 10% and 40%, more respectively. Until 1998 year there were 103 baby friendly hospitals in Brazil, with the majority of them located in the northeast area (68.1%). However, taking in accounting the number of 5650 hospitals linked to SUS in the country, less than 2.0% are baby friendly hospitals. On the basis of the experience and according with PNIAM data the implementation of the ten steps and the incentive to breastfeeding through baby friendly hospitals have resulted in a significant increase of breastfeeding incidence and duration in Brazil. PMID- 9852653 TI - [Clinical value of ambulatory monitoring of blood pressure]. PMID- 9852654 TI - [Multifunctional territories in human pathology]. PMID- 9852655 TI - [Perception on and registration of alcohol abuse and alcohol-related diseases at a university general hospital]. AB - BACKGROUND: The high frequency of alcohol related problems highlights the importance of its approach. The association of both the alcohol-abuse and alcohol related diseases has been pointed in the literature and are often the main reason for hospitalization. Physicians use different tools to detect abusive drinkers, and one of them is the CAGE-test, validated by Mayfield et al. in 1974. OBJECTIVES: To study the association between the result of the CAGE-test and the perception and registration of alcohol abuse by the medical staff and the nurses in charge of inpatients of Hospital de Clinicas de Porto Alegre, southern Brazil. MATERIAL AND METHOD: The design involved three phases. First, the application of the CAGE-test to the 385 eligible patients. Second, review of all 51 CAGE positive patients' records as well the controlled evaluation of 51 randomized CAGE-negative patients, to seek alcoholic habits and alcohol-related diseases. Third, interview with the medical staff to find out their perception about these subjects. RESULTS: We found 51 patients CAGE-positive (prevalence = 13%). When evaluated through the registration and through a standardized interview, medical staff and nurses were not aware of more than half of CAGE-positive patients. No significant differences were found between medical staff registration and perception; the prevalence of alcohol related diseases was the same for both: 26 and 27% (p = .861). The prevalence of alcohol related diseases was higher among CAGE-positive patients (p = .0003). CONCLUSIONS: In spite of the high prevalence among our patients, alcohol abuse is not pursued by our health teams as it should be. As for the future, there is reason to envision a continuing alcoholism rapprochement. PMID- 9852656 TI - [Obstruction of terminal ileum due to heterotopic gastric mucosa]. AB - Heterotopic gastric mucosa situated in the small bowel distal to the Treitz suspensory ligament is very rare, except in Meckel's diverticulum and in intestinal duplications. There are two forms of this disease, congenital and acquired. The former is secondary to inflammatory bowel disease. The main difference between these forms is histological, although determining diverse physiopathological aspects. A case of a 34 year old man with heterotopic gastric mucosa in the terminal ileum manifested by intestinal obstruction is reported. He was treated surgically by enterectomy of two small bowel segments, both reconstructed by primary suture. His postoperative course was remarkable. The histopathologic study showed a typical pattern of the acquired type because of the presence of antral the antral mucosa and intense fibrosis. That is probably related to intestinal tuberculosis, but was not histologically confirmed. Individual and family recent history of pulmonary tuberculosis corroborates the suspicion. This is a unique report in the literature, among 28 other heterotopic gastric mucosa situated in the jejunum and ileum. PMID- 9852657 TI - [Langerhans cell histiocytosis in anogenital region]. AB - The Langerhans cell histiocytosis (LCH) of the genital tract is rare, with only 48 cases related in the literature. There were reported only 2 cases in the anogenital region. We reported the third case of LCH in the anogenital region; patient was female, 31 years-old, caucasian and the diagnosis was confirmed by electron microscopic magnification. The treatment was local surgical excision and systemic chemotherapy. PMID- 9852658 TI - A comparison of temperature measurements using three ear thermometers. AB - Following anecdotal and research-based reports of inaccuracies encountered with the use of ear thermometers in patient care settings, this two-phase study was designed to compare the accuracy of three ear thermometers when used in a multioperator environment. The within-subject variation (limits of agreement) in ear temperature measurements obtained with the three ear thermometers in a multioperator condition by using an oral temperature reference standard ranged from +/- 1.25 degrees F to 1.85 degrees F. In multioperator, multisubject patient care environments using a pulmonary artery catheter core temperature as the reference standard, the limits of agreement for ear temperature measurements obtained with the three different ear thermometers ranged from +/- 2.19 degrees F to 2.85 degrees F. These results suggest that there is substantial variation in ear measurements and raise questions about the use of ear thermometers when there are multiple personnel taking temperatures, as occurs in hospital-based clinical practice environments. Handedness of the operator, position of the patient, and the ear used for measurement did not produce clinically significant variability in ear temperature measurements. PMID- 9852659 TI - Late-stage dementia and participation in therapeutic activities. AB - This study examined the participation and behavior of people with late-stage dementia during various therapeutic activities. Subjects were able to actively participate for 10 minutes or less 54% of the time. Caregivers were not responding to meet apparent resident needs 37.5% of the time. Active participation was significantly correlated with having more senses stimulated. In addition, as the length of resting time increased before a scheduled therapy, active participation significantly decreased. Subjects exhibited various behaviors during therapeutic activities. Behaviors were categorized as approaching and withdrawing, relaxing and tensing, mimicking, nonverbal connecting, and verbal connecting. PMID- 9852660 TI - Split-thickness skin graft donor site care: a quantitative synthesis of the research. AB - Split-thickness skin grafting (STSG) is a frequently used reconstructive technique but is associated with a large variation in practice. The purposes of this article are to integrate and synthesize the available empirical evidence regarding STSG donor site dressings, identify which dressings are associated with the best outcomes, and provide practice recommendations. This review of 33 studies found transparent film to be the best dressing for the care of STSG donor site wounds. Transparent film was associated with one of the fastest healing rates (9.47 days), a smooth epithelialized surface, a low infection rate (10 out of 394 patients or 3%), the least amount of pain (1.59 on 0 to 10 scale), and minimal cost ($.005 per square inch) when compared with other dressings. PMID- 9852661 TI - Recruitment barriers in rural community hospitals: a comparison of nursing and nonnursing factors. AB - Despite encouraging signs suggesting that many rural hospitals are experiencing less severe staff shortages, the challenge of successfully recruiting nurses to rural practice settings continues to be a major obstacle. Based on a 1993 survey of 164 directors of nursing (DONs) practicing in rural community hospitals, exploratory factor analysis (EFA) was used to cluster 21 DON-rated recruitment barriers into factors associated with nursing delivery, local community, and professional interaction. DONs, most notably those practicing in the smallest rural facilities (< or = 25 beds), reported community-related barriers such as spouse's employment as the most severe obstacles to successful registered nurse (RN) recruitment. However, upon examination conducted by using multiple regression analyses, only those factors related to nursing delivery and professional interaction were found to be statistically significant predictors of existing staff RN full-time equivalency vacancy rates. Given that barriers related to individual nursing staffs are likely to be far more amenable to change than those associated with rural environments, these findings offer encouragement to nursing administrators who are faced with the difficulties of attracting RNs to rural practice settings. PMID- 9852662 TI - Nursing intervention to increase the duration of breastfeeding. PMID- 9852663 TI - Utilization-focused integrative reviews in a nursing service. PMID- 9852664 TI - Syntax and morphology in Williams syndrome. AB - Williams syndrome (WS) is a neuro-developmental disorder which is characterized by an unusual fractionation of language abilities and other cognitive functions. We have investigated four cases of English-speaking subjects with WS, and we show that despite their low IQs the WS children's performance on syntactic tasks and on regular inflection is not impaired. Irregular inflection, however, is affected causing many errors. We also report results from studies investigating the same linguistic phenomena in children with specific language impairment. These children exhibit a different pattern of impairment, with relatively poor performance on syntactic tasks and regular inflection. We suggest a linguistic characterization of the morphosyntax in WS according to which WS subjects are impaired in accessing (particular kinds of) information from lexical entries, with their computational system for language appearing to be intact. We interpret the selective impairments found in WS and SLI as supporting the theoretical distinction between a computational system and an associative memory system for language. PMID- 9852665 TI - Mapping attractor fields in face space: the atypicality bias in face recognition. AB - A familiar face can be recognized across many changes in the stimulus input. In this research, the many-to-one mapping of face stimuli to a single face memory is referred to as a face memory's 'attractor field'. According to the attractor field approach, a face memory will be activated by any stimuli falling within the boundaries of its attractor field. It was predicted that by virtue of its location in a multi-dimensional face space, the attractor field of an atypical face will be larger than the attractor field of a typical face. To test this prediction, subjects make likeness judgments to morphed faces that contained a 50/50 contribution from an atypical and a typical parent face. The main result of four experiments was that the morph face was judged to bear a stronger resemblance to the atypical face parent than the typical face parent. The computational basis of the atypicality bias was demonstrated in a neural network simulation where morph inputs of atypical and typical representations elicited stronger activation of atypical output units than of typical output units. Together, the behavioral and simulation evidence supports the view that the attractor fields of atypical faces span over a broader region of face space that the attractor fields of typical faces. PMID- 9852666 TI - Mental rotation within linguistic and non-linguistic domains in users of American sign language. AB - American sign language (ASL) uses space itself to encode spatial information. Spatial scenes are most often described from the perspective of the person signing (the 'narrator'), such that the viewer must perform what amounts to a 180 degrees mental rotation to correctly comprehend the description. But scenes can also be described, non-canonically, from the viewer's perspective, in which case no rotation is required. Is mental rotation during sign language processing difficult for ASL signers? Are there differences between linguistic and non linguistic mental rotation? Experiment 1 required subjects to decide whether a signed description matched a room presented on videotape. Deaf ASL signers were more accurate when viewing scenes described from the narrator's perspective (even though rotation is required) than from the viewer's perspective (no rotation required). In Experiment 2, deaf signers and hearing non-signers viewed videotapes of objects appearing briefly and sequentially on a board marked with an entrance. This board either matched an identical board in front of the subject or was rotated 180 degrees. Subjects were asked to place objects on their board in the orientation and location shown on the video, making the appropriate rotation when required. All subjects were significantly less accurate when rotation was required, but ASL signers performed significantly better than hearing non-signers under rotation. ASL signers were also more accurate in remembering object orientation. Signers then viewed a video in which the same scenes were signed from the two perspectives (i.e. rotation required or no rotation required). In contrast to their performance with real objects, signers did not show the typical mental rotation effect. Males outperformed females on the rotation task with objects, but the superiority disappeared in the linguistic condition. We discuss the nature of the ASL mental rotation transformation, and we conclude that habitual use of ASL can enhance non-linguistic cognitive processes thus providing evidence for (a form of) the linguistic relativity hypothesis. PMID- 9852667 TI - A phoneme effect in visual word recognition. AB - In alphabetic writing systems like English or French, many words are composed of more letters than phonemes (e.g. BEACH is composed of five letters and three phonemes, i.e./biJ/). This is due to the presence of higher order graphemes, that is, groups of letters that map into a single phoneme (e.g. EA and CH in BEACH map into the single phonemes /i/ and /J/, respectively). The present study investigated the potential role of these subsyllabic components for the visual recognition of words in a perceptual identification task. In Experiment 1, we manipulated the number of phonemes in monosyllabic, low frequency, five-letter, English words, and found that identification times were longer for words with a small number of phonemes than for words with a large number of phonemes. In Experiment 2, this 'phoneme effect' was replicated in French for low frequency, but not for high frequency, monosyllabic words. These results suggest that subsyllabic components, also referred to as functional orthographic units, play a crucial role as elementary building blocks of visual word recognition. PMID- 9852668 TI - Predictive saccade behavior is enhanced in schizophrenia. AB - In this study, we investigated the ability of schizophrenic patients to produce predictive saccades in response to a visual target moving with predictable timing and location. The performance of visually guided saccades and predictive saccades (gain and latency) were analyzed in a group of 12 schizophrenic patients as compared with a group of ten control subjects. Our main finding was an enhancement of the predictive tracking ability in schizophrenics. In particular, the predictive tracking built-up and became installed as a steady pattern of highly anticipatory saccades (in terms of percentage of negative latency saccades) much faster in the schizophrenic group than in the control group. These data are discussed in terms of an involvement of the fronto-striatal system in this specific enhancement of predictive saccade behavior in schizophrenia. PMID- 9852669 TI - Conditional reasoning, causality, and the structure of semantic memory: strength of association as a predictive factor for content effects. AB - Available evidence indicates that responses to conditional inferences using concrete causal premises is affected by the relative number of available alternate causes (Cummins, D.D., 1995. Memory and Cognition 23 (5), 646-658). We propose that another important factor that may influence the kinds of inferences made to causal conditionals is the relative strength of association between such causes and the consequent term. We present a study with adult participants that examines the effect of strength of association on performance on a conditional reasoning task using causal premises for which there exist one highly associated potential cause for the given consequent term. We predicted that adults would produce a greater proportion of biconditional responses to invalid forms with strongly associated premises than weakly associated ones, while valid forms would not be affected by strength of association. The results are consistent with this hypothesis. PMID- 9852670 TI - Anticipation in Japanese families with schizophrenia. AB - The identification of anticipation in schizophrenia is a recent focus in the genetic epidemiology of schizophrenia, although it involves some controversial methodological issues. We explored the evidence of anticipation among 44 Japanese two-generation pairs with schizophrenia found by reviewing nine years of admission records (1986-1994) at the Department of Neuropsychiatry, Nagasaki University Hospital and Michino-o Hospital, Nagasaki. The 44 pairs consisted of 27 two-generation pairs of first-degree relatives group (FDRG) and 17 pairs of second-degree relatives group (SDRG). On pairwise comparison and a life table analysis, the age at onset (AO) was significantly earlier in the lower generation (G2) than in the upper generation (G1) in all of the pairs and in the two subgroups, FDRG and SDRG. Earlier AO was shown in G2 even after minimizing some statistical biases for the study of anticipation in schizophrenia. A significant earlier mean AO was found in G2 even when a cohort effect was controlled for. There was no marked difference in AO between paternal and maternal transmission. These results provide further evidence for epidemiological anticipation, suggesting biological anticipation such as the involvement of trinucleotide repeats expansion in G2. The limitations of the study are also discussed. PMID- 9852671 TI - New dominant syndrome of microcephaly, facial abnormalities, micromelia, and mental retardation. AB - We report on three brothers, aged 6, 3, and 2 years, with a hitherto undescribed combination of microcephaly, facial abnormalities, micromelia, and mild mental retardation. Their facial abnormalities included a forehead with bitemporal constriction, upslanting palpebral fissures, synophrys, a short nose with anteverted nostrils, a short columella, a cupid bow-shaped, thin vermilion border of the upper lip, and micrognathia. Their mother had similar clinical manifestations, but was of normal intelligence. The disease was apparently transmitted in a dominant fashion. PMID- 9852672 TI - Infrequent genetic alterations of the PTEN gene in Japanese patients with sporadic prostate cancer. AB - Prostate cancer is a major cause of cancer death among elderly men in America, Europe, and Japan. However, the molecular mechanism of carcinogenesis is not yet well characterized. Frequent loss of heterozygosity (LOH) on chromosome 10q was reported in prostate cancer, and a candidate tumor suppressor gene, PTEN, was isolated on chromosome band 10q23.3. To investigate the genetic alterations of PTEN, we examined 45 primary prostate cancer specimens. LOH at the PTEN locus was observed in two (11.1%) of 18 tumors. However, no mutations were observed in any of the primary prostate cancers. These data suggest that mutation of the PTEN gene does not play a major role in prostate carcinogenesis of Japanese patients. PMID- 9852673 TI - Mutation analysis of the phenylalanine hydroxylase gene and its clinical implications in two Japanese patients with non-phenylketonuria hyperphenylalaninemia. AB - We describe a mutation analysis for the phenylalanine hydroxylase gene and the clinical outcome of two Japanese patients with non-phenylketonuria (PKU) hyperphenylalaninemia (serum phenylalanine level below 600 mumol/l under a free diet) detected by a mass-screening program. Single strand conformation polynorphism analysis and direct sequencing of their genomic DNAs revealed that non-PKU hyperphenylalaninemia resulted from compound heterozygosity for a mutation causing classical PKU and a mutation with a milder effect on phenylalanine hydroxylase activity. The mutations were R241C and R243Q in exon 7, and R413P in exon 12. The mutation genotypes of the two patients were R241C/R243Q and R241C/R413P. It has been demonstrated that homozygosity for the R243Q or R413P mutation is associated with a severe phenotype of PKU and low in vitro expression activity. In contrast, the R241C mutation has much less effect on phenylalanine hydroxylase activity. The metabolic consequence of each variant allele was confirmed by a phenylalanine loading test in the patients and their parents. The patients achieved normal results in all intellectual and neurologic tests. Brain magnetic resonance imaging revealed no abnormalities. The dietary restriction was continued until 10 years of age in order to maintain the serum phenylalanine level below 400 mumol/l. The genetic analysis to distinguish non PKU hyper-phenylalaninemia from classical PKU helps to determine the principles of dietary management in the early infantile period. PMID- 9852674 TI - Molecular diagnosis of patients with beta-thalassemia major in central Taiwan by amplified created restriction site analysis. AB - beta-Thalassemia, a hematologic disorder characterized by the deficiency or the absence of beta-globin production, is the most widespread inherited disorder in the world; it is also common in Taiwan. We studied 38 patients in central Taiwan with beta-thalassemia major, using amplified created restriction site analysis for detection. On analysis, six different point mutations were found among 76 chromosomes, of which 32 chromosomes (42.1%) had a C to T substitution at nucleotide 654, 30 (40%) had frameshift codons 41/42 with four nucleotides (TCTT) deletion, 7 (9.2%) had an A to T substitution at codon 17, 3 (3.9%) had frameshift codons 71/72 (insertion of A), 2 (2.6%) had an A to G substitution at position -28, and 2 (2.6%) had frame-shift codons 27/28 (insertion of C). The first two mutations accounted for 62 of the 76 beta-thalassemia mutations in this study. As to mutations in each individual with beta-thalassemia major, the incidence of compound heterozygotes of two different mutations was higher than that of homozygotes of a single mutation (60% vs 40%). Compound heterozygotes of C to T substitution at nucleotide 654 of IVS-2 and frameshift codons 41/42 with four-nucleotide deletion was the most common pattern of beta-thalassemia mutations in each individual (23.7%). Our results were unique compared with those from similar studies performed in southern China. Frequencies of beta-thalassemia mutations found in the current study were assessed and compared with frequencies found in previous studies conducted in northern and southern Taiwan. PMID- 9852675 TI - Mitochondrial DNA mutations in Japanese patients with optic neuropathy unassociated with a mutation at nucleotide position 11,778. AB - We examined for mitochondrial DNA (mtDNA) mutations at nucleotide positions(nt) 3460, 14,484, 9438, 9804, and 15,257 in ten Japanese patients with idiopathic optic neuropathy unassociated with a mutation at nt11,778. The mtDNAs were amplified by polymerase chain reaction (PCR), the products were digested with restriction enzymes, and the sizes of the fragments were analyzed on 8% polyacrylamide gel. Of the ten patients, one had an mtDNA mutation at nt3460 and another patient had a mutation at nt14,484. We suggest that mtDNA mutations in Japanese patients with optic neuropathy unassociated with a mutation at nt11,778 should be further investigated. PMID- 9852676 TI - Further evidence for a major ancient mutation underlying myotonic dystrophy from linkage disequilibrium studies in the Japanese population. AB - The myotonic dystrophy (DM) mutation is an unstable (CTG)n repeat, present at a copy number of 5-37 repeats on normal chromosomes but amplified to 50-3000 copies on DM chromosomes. Previous findings in Caucasian populations of a DM founder chromosome raise a question about the molecular events involved in the expansion mutation. To investigate whether a founder chromosome for the DM mutation exists in the Japanese population, we genotyped families using polymorphic markers near the (CTG)n repeat region and constructed haplotypes. Six different haplotypes were found and DM alleles were always haplotype A. To find an origin of the (CTG)n repeat mutation and to investigate the mechanism of the expansion mutation in the Japanese population we have studied 90 Japanese DM families comprising 190 affected and 130 unaffected members. The results suggest that a few common ancestral mutations in both Caucasian and Japanese populations have originated by expansion of an ancestral n = 5 repeat to n = 19-37 copies. These data support multistep models of triplet repeat expansion that have been proposed for both DM and Friedreich's ataxia. PMID- 9852677 TI - Five familial hypercholesterolemic kindreds in Japan with novel mutations of the LDL receptor gene. AB - In the course of investigations of familial coronary artery disease in Hokkaido, the northern island of Japan, we identified five families in which multiple members showed elevated plasma levels of low-density lipoprotein (LDL) cholesterol. To determine the genetic etiology of their lipoprotein abnormalities, we screened DNA samples from these families for mutations in all 18 exons and the exon-intron boundaries of the LDL receptor (LDLR) gene. Novel point mutations were identified in each family: (1) a C-to-A transversion at nucleotide 285, causing a nonsense mutation at codon 74, in eight members of family A; (2) a G-to-A transition at nucleotide 1136, causing substitution of Tyr for Cys at codon 358, in six members of family B; (3) a C-to-T transition at nucleotide 1822, causing substitution of Ser for Pro at codon 587, in five members of family C; (4) a one-base insertion of G to a five-G stretch at nucleotides 1774-1778 (codons 571-572), causing a frameshift, in six members of family D; and (5) a one-base deletion of T at nucleotide 1963-1964 (codon 634), causing a frameshift, in three members of family E. Through the molecular genetic approach a total of 28 individuals in these families were diagnosed unequivocally as heterozygous for the respective LDLR mutations. This method also helped us to diagnose familial hypercholesterolemia, or to exclude from carrier status, 11 children with borderline high cholesterol levels. PMID- 9852678 TI - Isolation and mapping of a novel human kidney- and liver-specific gene homologous to the bacterial acetyltransferases. AB - Using the differential display method to detect tissue-specific genes, we isolated a novel human gene, designated TSC501, that is expressed in kidney and liver. The cDNA contained an open reading frame of 681 nucleotides encoding 227 amino acids. The predicted product showed homologies in amino acid sequence to three different bacterial acetyltransferases, enzymes that are involved in drug resistance. Radiation hybrid mapping localized this gene to human chromosome bands 2p12-p13.1. PMID- 9852679 TI - Novel missense mutation resulting in the substitution of tyrosine by cysteine at codon 597 of the type X collagen gene associated with Schmid metaphyseal chondrodysplasia. AB - Schmid metaphyseal chondrodysplasia (SMCD) is one of the most common forms of the osteochondrodysplasias. Mutations or deletions in the COL10A1 gene that encodes type X collagen have been shown to cause this disorder. Most of the gene mutations and deletions are located in the non-collagenous carboxy (C)-terminal (NC1) domain. We describe a novel missense mutation in a patient with SMCD that leads to the substitution of Tyr at codon 597 by Cys in the NC1 domain. Sequence analysis indicated that the proband was heterozygous for the mutation. Her parents were homozygous for the normal sequence, indicating the de-novo occurrence of this mutation. PMID- 9852680 TI - Cloning and characterization of the murine P2XM receptor gene. AB - We have isolated the murine counterpart of the human P2XM gene (mP2XM), a P2X purinoceptor that is expressed predominantly in skeletal muscle. The mP2XM gene, consisting of 12 exons that span 10 kb of genomic DNA, encodes a 379-amino-acid product with 83% identity to the human homologue. Two potential transmembrane domains (M1 and M2) are present in the predicted product, and a segment resembling the H5 region of voltage-gated ion channels is completely conserved between human and mouse P2XM proteins. Predominant expression of mP2XM in murine skeletal muscle was confirmed by Northern-blot analysis, and a low level of expression was detectable in lung tissue, although human lung does not express P2XM. PMID- 9852681 TI - Cloning, expression analysis, and chromosomal localization of HIP1R, an isolog of huntingtin interacting protein (HIP1). AB - Huntington disease (HD) is an inherited neurodegenerative disorder which is associated with CAG expansion in the coding region of the gene for huntingtin protein. Recently, a huntingtin interacting protein, HIP1, was isolated by the yeast two-hybrid system. Here we report the isolation of a cDNA clone for HIP1R (huntingtin interacting protein-1 related), which encodes a predicted protein product sharing a striking homology with HIP1. RT-PCR analysis showed that the messenger RNA was ubiquitously expressed in various human tissues. Based on PCR assisted analysis of a radiation hybrid panel and fluorescence in situ hybridization, HIP1R was localized to the q24 region of chromosome 12. PMID- 9852682 TI - Isolation, tissue expression, and chromosomal assignment of a novel human gene which encodes a protein with RING finger motif. AB - We identified a novel gene encoding a RING finger (C3HC4-type zinc finger) protein from a human neuroblastoma full-length enriched cDNA library. This cDNA clone consists of 1919 nucleotides with an open reading frame of a 485-amino acid protein. From reverse transcription (RT)-polymerase chain reaction (PCR) analysis, the messenger RNA was ubiquitously expressed in various human adult tissues. The chromosomal location of the gene was determined on the chromosome 6p21.3 region by PCR-based analyses with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid mapping panel. PMID- 9852683 TI - Chromosomal assignment of the gene for human DNA-PKcs interacting protein (KIP) on chromosome 15q25.3-q26.1 by somatic hybrid analysis and fluorescence in situ hybridization. AB - We report the chromosomal location of the gene for DNA-PKcs interacting protein KIP. Based on fluorescence in situ hybridization and polymerase chain reaction (PCR)-based analyses with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid mapping panel, this gene was mapped to q25.3-q26.1 region on chromosome 15. PMID- 9852684 TI - A highly polymorphic CA repeat marker at the human tumor necrosis factor alpha (TNFA alpha) locus. AB - Tumor necrosis factor alpha (TNF alpha) in activated monocytes exerts cytotoxic activity and has a variety of other biological effects. We isolated a polymorphic dinucleotide (CA) repeat sequence from a genomic clone containing the gene located at 6p21.3. High heterozygosity (0.80) makes this polymorphism a useful marker in the genetic study of disorders affecting immunological response and cell differentiation. PMID- 9852686 TI - A HhaI/BstUI polymorphism in a novel gene at human chromosome 11p15.5. AB - We found a HhaI/BstUI polymorphism in the 3' untranslated region of a novel gene which was localized to 11p15.5. This region is one of prominent imprinting domains and contains multiple imprinted genes, such as H19, IGF2, KVLQT1, and p57KIP2, which suggests that regional factors might contribute to the imprinting. This polymorphism will be useful in the allelic analysis of expression and methylation of the novel gene. PMID- 9852685 TI - Isolation and radiation hybrid mapping of a dinucleotide repeat polymorphism at the human calcium-sensing receptor (CASR) locus. AB - Calcium-sensing receptor (CASR) in parathyroid gland regulates calcium homeostasis by sensing decreases in extracellular calcium levels and effecting an increase in secretion of parathyroid hormone. A polymorphic dinucleotide (CA) sequence was isolated from a genomic clone containing the human CASR gene and was mapped to 3q13.3-q21. This polymorphism will be useful in the genetic study of disorders affecting calcium metabolism, such as hypercalcemia, hypocalcemia, osteoporosis, hyperparathyroidism, and hypoparathyroidism. PMID- 9852688 TI - Isolation of a polymorphic CA repeat sequence at the human progesterone receptor (PGR) locus. AB - We isolated a polymorphic dinucleotide (CA) repeat sequence from a genomic clone containing the human progesterone receptor (PGR) gene. This polymorphism will be a useful marker in the genetic study of disorders affecting female endocrine systems, such as progesterone resistance and breast, uterine, and ovarian cancers. PMID- 9852687 TI - A GT dinucleotide repeat polymorphism in intron 1 of the H-cadherin (CDH13) gene. AB - A polymorphic dinucleotide (GT) repeat sequence was isolated from a genomic clone containing the human H-cadherin (CDH13) gene at 16q24.1-q24.2. This polymorphic marker will be a useful tool in the genetic study of various cancers. PMID- 9852689 TI - A highly polymorphic CA repeat marker at the human interleukin 6 receptor (IL6R) locus. AB - A polymorphic dinucleotide (CA) sequence was isolated from a genomic clone containing the human interleukin 6 receptor (IL6R) gene. High heterozygosity (0.81) makes this polymorphism a useful marker in the genetic study of disorders affecting the inflammation process and bone resorption. PMID- 9852690 TI - [Effects of cardiac output on blood and tissue pH during general anesthesia with constant ventilation]. AB - We investigated the effects of cardiac output on blood and tissue pH in 106 adult patients undergoing cardiac or non-cardiac surgery under general anesthesia. After anesthetic induction, the minute ventilation volume was kept constant at 10 ml.kg-1 x 10 cycles.min-1. A pulmonary artery catheter and a nasogastric tube incorporating a tonometer were inserted. During surgery, cardiac index (CI), pH, Pco2, BE, So2 and Hb of arterial and mixed venous blood as well as gastric intramucosal pH (pHi) were measured simultaneously. Oxygen uptake index (Vo2I) and blood CO2 contents were calculated. The measurements were repeated every 10 to 20 minutes during surgery or during the prebypass period. Two patients with preoperative cardiogenic shock were excluded from data analysis because of development of severe acidosis and 624 sets of data from 104 patients were analyzed. Arterial and mixed venous pH correlated negatively with CI. Blood Pco2 and base excess (BE) correlated positively and negatively, respectively, with CI. Blood lactate concentration measured 142 times in the last 30 patients correlated positively with CI. Vo2I correlated positively with CI and Paco2 correlated positively with Vo2I. Veno-arterial differences in Pco2 and Cco2 correlated negatively with CI. Due to the difference, Caco2 correlated positively with CI, while Cvco2 did not correlate with CI. pHi correlated negatively with CI but only marginally. By multiple regression analysis, pHi was not affected significantly by CI, while it showed positive correlation with pHa, Hb, Sao2 and negative correlation with blood temperature. When cardiac output increased, blood pH decreased due to increased Pco2 and decreased BE. An increase in Paco2 might result from both an increase in Vo2 or Vco2 and decreased ventilation-to perfusion ratio. A decrease in BE might result from increased washout of acids (e.g. lactate) from the tissue to the central circulation. In contrast to blood pH, pHi or tissue pH was not affected significantly by cardiac output unless patients were in cardiogenic shock. PMID- 9852691 TI - [Effect of ketamine on the Ca(2+)-related functions of skinned skeletal muscle fibers from the guinea pigs]. AB - To determine whether ketamine has a possibility to predispose a patient to malignant hyperthermia (MH), the author investigated the effects of ketamine on the intracellular calcium kinetics with the skinned fiber technique in skeletal muscle cells of guinea pigs. Intracellular calcium kinetics including calcium induced calcium release (CICR) from sarcoplasmic reticulum (SR), calcium uptake into SR, and sensitivity of contractile proteins for calcium were assessed by using calcium concentration-response relationships. The effects of ketamine on the calcium concentration-response curves, which were constructed by changing calcium concentration from 10(-7) to 10(-4.5) M, were determined with ketamine concentrations of 0.1, 0.3, 1.0 and 3.0 mM. The author found that the rate of CICR was not altered with any ketamine concentration, while initial rate of calcium uptake into SR decreased and calcium rate (pCa)-tension curves shifted leftward with ketamine concentration of 3 mM at low concentrations of Ca2+ (below pCa 5.5). Thus, ketamine is not associated directly with occurrence of MH because ketamine has no significant effects on CICR. However, an increase in intracellular calcium concentration due to depressed calcium uptake into SR and an increased sensitivity of contractile proteins for calcium may lead to symptoms and signs like MH. PMID- 9852692 TI - [The inhibition of lidocaine metabolism by various barbiturates in rat hepatic microsome]. AB - To evaluate the effects of various barbiturates on lidocaine metabolism by cytochrome P-450 (P-450), enzyme kinetics were analyzed in an in vitro study using rat hepatic microsomes. Phenobarbital, amobarbital, hexobarbital, pentobarbital, and thiamylal showed the mixed type inhibition of lidocaine metabolism with inhibition constants being 4.89, 1.08, 2.76, 0.77 and 0.65 mM, respectively. Same as lidocaine, all barbiturates used in the present study, corresponding to binding with P-450, induced the I type of spectral change of P 450. Since these did not affect cytochrome C reductase activity, it was suggested that this inhibition of lidocaine metabolism in hepatic microsomes may have been caused by the reduction of activity on P-450 by the barbiturates. PMID- 9852693 TI - [Evaluation of preincisional mexiletine administration to alleviate postoperative pain]. AB - Mexiletine, an antiarrhythmic agent, was preincisionally administered intravenously for the purpose of reducing postoperative pain. Twenty-eight female patients for mastectomy were studied. The patients were divided into three groups. Group 1 received no mexiletine. Group 2 received bolus administration of mexiletine 1 mg.kg-1 with additional continuous administration of 1 mg.kg-1.hr-1 for 75 minutes. Group 3 received bolus administration of mexiletine 2 mg.kg-1. The requirement of butorphanol as a postoperative analgesic within 1 hour after mastectomy in Group 3 was significantly lower than that in Group 1 (P < 0.05), but butorphanol requirement in Group 2 was not significantly lower than that in Group 1. Plasma mexiletine concentration was slightly higher in Group 3 (1.7 micrograms.ml-1) than that in Group 2 (1.0 microgram.ml-1) immediately after the intravenous mexiletine administration, although there was no significant difference. The results indicate that mexiletine 2 mg.kg-1 as preoperative bolus administration maintains its plasma concentration above 1.7 micrograms.ml-1, and is clinically effective for reducing the postoperative pain after mastectomy. PMID- 9852694 TI - [Comparison of propofol and pentazocine combined with thiamylal for laryngeal mask insertion]. AB - We evaluated the combination of pentazocine and thiamylal as induction agents for laryngeal mask airway (LMA) insertion and compared this with propofol. Ninety four patients, ASA grade 1 or 2, were randomly assigned to one of four induction groups as follows; group P: propofol 2.5 mg.kg-1, group p 0.3: pentazocine 0.3 mg.kg-1 followed by thiamylal 5 mg.kg-1, group p 0.6: pentazocine 0.6 mg.kg-1 followed by thiamylal 5 mg.kg-1, and group T: thiamylal 5 mg.kg-1. In group T (n = 5), insertion of LMA was impossible due to inadequate anesthesia. In contrast, good and acceptable conditions for LMA insertion were obtained in 85.2%, 86.7%, and 96.9% of group P, p 0.3, and p 0.6, respectively. Apneic interval, endtidal CO2 and arterial CO2 were significantly greater in pentazocine groups than in propofol group (group P < p 0.3 < p 0.6). Systolic pressure decreased after induction in all groups. Decreases in systolic and diastolic pressure were significantly greater in group P. Heart rate did not show any significant change. We conclude that the induction with the combination of pentazocine and thiamylal provides suitable conditions for LMA insertion with more stable hemodynamics compared with propofol. Doses of 0.3 mg.kg-1 seem to be desirable for LMA insertion. PMID- 9852695 TI - [Anesthesia induction for laryngeal mask insertion--comparison among sevoflurane, isoflurane and propofol]. AB - We compared the patient's response to laryngeal mask (LM) insertion and hemodynamics among three anesthesia induction methods; S group used sevoflurane and slowly increased to 5% in 50% nitrous oxide; I group used isoflurane and slowly increased to 3% in 50% nitrous oxide; and P group used 2.5 mg.kg-1 propofol with 0.2 microgram.kg-1 fentanyl. Thirty patients, 35 to 65 years, for elective mastectomy were the subjects of study in each of the three groups. Preanesthetic medication was composed of i.m. injection of 0.5 mg atropine and 5 mg midazolam 30 min. prior to the induction. The number of patients with difficult insertion or demonstrating body movement or gagging in each group was in the order of P group > I group > S group. Blood pressure (BP), heart rate (HR) and rate pressure product (RPP) in the P group were significantly lower than those in the other two groups. I group showed significant increase in BP, HR and RPP. It was concluded that sevoflurane enabled the most smooth insertion of LM among the three methods of anesthesia with the least hemodynamic change. PMID- 9852696 TI - [Current topics in the regulation of prostanoids-1. Inducible cyclooxygenase COX 2 and selective blockers]. AB - Cyclooxygenases (COX) are prostaglandin synthases and are the main therapeutic targets for non-steroidal anti-inflammatory drugs. Recently it has been established that apart from the constitutive isoform, an inducible isoform of this enzyme is upregulated after injurious stimuli. The structures of the genes encoding these enzymes and protein structures have been determined, and with the utilization of knockout mice, the physiological properties of each isozyme are being revealed. COX-1 is responsible for the production of physiological levels of prostanoids, whereas COX-2 is upregulated during inflammatory states and produces prostanoids responsible for the generation of fever, pain or other inflammatory responses. Selective COX-2 blockers may be a safe and useful alternative in the treatment of inflammatory disorders, pain, and fever. PMID- 9852697 TI - [Pulmonary edema due to acute airway obstruction immediately after tracheal extubation]. AB - A 33-year-old male was scheduled for tonsillectomy and pharyngoplasty due to sleep apnea syndrome. The intubation was uneventful following induction with thiamylal and vecuronium. Anesthesia was maintained with O2-N2O-sevoflurane. No complications were observed during the 90 min operation. After the termination of the anesthesia, a hyperadrenergic state was observed: arterial pressure and heart rate rose to 230/135 mmHg and 135 bpm, respectively. Immediately after extubation, he developed dyspnea with tracheal tag and stridor, and became cyanotic despite the use of a simple oxygen mask and assisted ventilation. Laryngospasm was suspected. The patient was reintubated and suctioned; pink, frothy sputum was not obtained. Arterial blood gases 5 minutes after reintubation revealed a pH of 7.24, Pao2 86 mmHg (FIo2 1.0), and Paco2 54 mmHg. Chest X-ray 30 minutes after reintubation revealed bilateral diffuse alveolar infiltration. The diagnosis was interstitial pulmonary edema. The patient was ventilated mechanically by applying a positive end-expiratory pressure of 5cm H2O, and furosemide and dopamine were administered intravenously. The patient was extubated the next day, and discharged from hospital ten days later. We considered that the lung edema was induced by the severe negative pressure generated by inspirating against a closed upper airway, as well as by the hyperadrenergic state and severe hypoxemia observed during and after extubation. PMID- 9852698 TI - [Pulmonary fat embolism during bipolar hip endoprosthesis]. AB - Two patients had fatal fat pulmonary embolism during bipolar hip endoprosthesis. Two women, 71-year-old and 76-year-old, with femoral neck fracture underwent bipolar hip endoprosthesis under combined lumbar epidural/general anesthesia. Soon after the placement of bone cement and a femoral stem with a bipolar endoprosthesis in the femoral shaft, the patients developed circulatory collapse. Immediate cardiopulmonary resuscitation did not restore adequate circulation and the patients died 3.5 hours and 1 hour thereafter, respectively. Pathological examination revealed the presence of fat particles within the entire pulmonary arteries, arterioles and capillaries. Main cause of the present pulmonary fat embolism may be an increase in the intramedullary pressure during cement pressurization and femoral stem placement. Anesthetists have to be aware of occurrence of this serious syndrome during bipolar hip endoprosthesis. For earlier detection of this serious complication we recommend to monitor central venous pressure and pulmonary artery pressure in patients undergoing bipolar hip endoprosthesis. PMID- 9852699 TI - [Surface-induced mild hypothermia anesthesia for hepatectomy in a patient with a giant hemangioma of the liver (Kasabach-Merritt syndrome)]. AB - A 63-year-old woman presenting with thrombocytepenia and signs of intravascular coagulation (prothronbin time, 59%: FDP, 100 micrograms.ml-1) due to a giant hemangioma of the liver (Kasabach-Merritt syndrome) was scheduled for the resection of the right lobe of the liver. In order to protect the liver on occlusion of the right hepatic artery and portal vein, we induced mild hypothermia technique with vasodilation and surface cooling by convecting warming device together with hepatoprotective agents of PGE1 and ulinastatin. Severe, acute massive bleeding occurred due to the injury of the middle hepatic vein and from the resected surface of the liver. Her rectal temperature was 31.9 degrees C during massive bleeding. Her hemoglobin decreased to 3.9 g.dl-1. Total estimated blood loss was 22,000 ml. The weight of the resected liver was 2.5 kg. The maximum postoperative levels of T-Bil and GOT were 2.47 mg.dl-1 and 171 IU.l-1, respectively. The liver seemed to have been well preserved and no other complications were observed. The hemangioma was completely removed by excision of the right lobe of the liver. Subsequently, all coagulation parameters returned to normal, indicating a complete reversibility of the coagulopathy. Surface-induced mild hypothermia is a useful and valuable method for protecting the liver during severe massive bleeding. PMID- 9852700 TI - [Effect of propofol as an agent for anesthetic induction on pituitary adrenocortical function during anesthesia and surgery]. AB - Effect of propofol as an agent for anesthetic induction on plasma levels of cortisol, beta-endorphin-like immunoreactivity (beta-ELI), growth hormone (GH) and prolactin were evaluated in 20 non-abdominal surgical patients ranged in ages from 19 to 64 years. Anesthesia was induced with either intravenous propofol 2 2.5 mg in ten patients or intravenous thiopental 4-5 mg in the remaining 10 patients as the control group, and succinylcholine was administered intravenously to facilitate tracheal intubation. Enflurane-nitrous oxide-oxygen was then given to maintain anesthesia in all the patients of both groups. Plasma cortisol levels decreased slightly with anesthesia in the propofol group, but they increased slightly after anesthetic induction in the control group. Therefore they were significantly lower in the propofol group than those in the control group. They tended to increase gradually during surgery and reached the peak value after the emergence from anesthesia in both groups. Plasma beta-ELI levels were unchanged with anesthesia alone in the patients of both groups. They tended to increase gradually during surgery and reached the peak value after the emergence from anesthesia in both groups. Plasma GH levels were not affected with anesthesia, but they increased slightly during surgery in both groups. Plasma prolactin levels increased significantly during anesthesia and surgery in both groups, and they decreased after the emergence from anesthesia but were still significantly higher than the preanesthetic values in both groups. The authors' findings suggest that effects of propofol as an agent for anesthetic induction on pituitary-adrenocortical function during anesthesia and surgery are comparable to those of thiopental, and that propofol does not exert inhibitory effect on pituitary-adrenocortical function during anesthesia and surgery. PMID- 9852701 TI - [Hemorrhagic shock during laparoscopic cholecystectomy detected by transesophageal echocardiography]. AB - A 47-year-old man was scheduled for laparoscopic cholecystectomy under general anesthesia supplemented with epidural anesthesia. A direct arterial line and a transesophageal echocardiogram probe were inserted before surgery. Anesthesia was maintained with nitrous oxide and isoflurane but without epidural anesthesia. Severe hypotension occurred about 30 minutes after introducing pneumoperitoneum but surgeons denied massive bleeding in the operative field. Although this made us difficult to diagnose the incident as massive bleeding or pulmonary air embolism (PAE), a collapsed heart was detected by transesophageal echocardiography (TEE). Its end-diastolic diameter of the left ventricle was reduced to 20 mm and left ventricular end-systolic cavity obliteration was demonstrated. We could easily diagnose the decrease of blood volume due to PAE using TEE. PMID- 9852702 TI - [Intraoperative transient incomplete left bundle branch block in a patient with left axis deviation in pre-anesthetic electrocardiogram]. AB - We encountered a case of transient incomplete left bundle branch block (TILBBB) during standard mastectomy under general anesthesia. The patient was a 40 year old female (70 kg, 164 cm) without any abnormalities on preanesthetic examinations except -61 degrees left axis deviation in exercise electrocardiogram. Adriamycin 20 mg was administered preoperatively. After the skin incision, heart rate increased from 104 min-1 to 130 min-1 and the cardiac axis gradually rotated leftward with increasing Q wave depth on leads I and aVL. We diagnosed this as blockade of the anterior branch in the left bundle branch. After the administration of fentanyl (0.2 mg) and sevoflurane (3%), the heart rate decreased to 105 min-1 and the electrocardiogram returned to the initial wave form. This anesthetic course indicated that adriamycin had slightly damaged the cardiac muscle and inadequate anesthesia had caused tachycardia and transient left bundle branch block. Left axis deviation on preoperative exercise electrocardiogram suggests that the left bundle branch can easily be blocked with an increasing heart rate. Adequate depth of anesthesia would have prevented the increase in heart rate and abnormality in the cardiac conduction process. PMID- 9852703 TI - [Anesthetic management of total en bloc spondylectomy]. AB - We analyzed the anesthetic management of 20 patients with total en bloc spondylectomy (TES) for solitary spinal metastases. Anesthesia was maintained with isoflurane, nitrous oxide in oxygen, and fentanyl in all patients. The duration of anesthesia was 733 minutes, and the amount of bleeding was 5371 ml on the average. During operation, hypovolemic shock occurred in 5 cases and huge hemothorax in 4 cases. Postoperative, huge hemothorax occurred in 6 cases and severe pneumonia in 3 cases. Maintenance of hemodynamic and respiratory function is important in perioperative management of TES. PMID- 9852704 TI - [A newly designed parallel connecting tube makes feasible high speed blood transfusion with a transfusion pump]. AB - Use of a transfusion pump (BP 101, Terumo, Tokyo) makes it feasible to obtain a stable and almost constant ejection volume at a flow rate of 99 ml.min-1, with no untoward effects of the intravenously placed needle and the microfilter located in the circuit. An air sensor ensures that the pump will cease operation immediately and automatically if an air bubble occurrs in the circuit. When a blood bag is conventionally connected to a connecting tube, at the maximum flow rate, one must set up a new blood bag within a few minutes, and in emergency situations with a shortage of hands, this would be unduly troublesome. When 5-7 blood bags (400 ml) is connected to 5-7 parallel connecting tubes, the pump will continuously eject blood approximately to 2000 ml at 99 ml.min-1, without replacing the blood bags. PMID- 9852705 TI - [A survey of opioid use in preanesthetic medication]. AB - A mailing survey was carried out to assess the current practice of sedative premedication in anesthesia. Questionnaires were sent by mail to 77 university hospitals. We especially evaluated use of opioids for premedication. Sixty-one percent (n = 47) of the hospitals answered these questionnaires and 31 percent (n = 15) of them were using opioids for the preanesthetic medication. Pethidine was used most frequently for premedication of all opioids, and morphine was often used for the premedication before cardiac surgery. All opioids were administrated intramusculary both in adults and children. PMID- 9852706 TI - The diversity of peripheral nervous system disorders in Sjogren's syndrome. PMID- 9852707 TI - Primary Sjogren's syndrome presenting as autonomic neuropathy. Case report. AB - Abnormalities of the autonomic nervous system have been described in several connective tissue diseases, but the relation with primary Sjogren's syndrome is unclear. This report describes a patient with primary Sjogren's syndrome who presented with severe autonomic failure. The present knowledge on dysfunction of the autonomic nervous system in primary Sjogren's syndrome and other connective tissue diseases is shortly reviewed. PMID- 9852708 TI - Non-conventional mechanical ventilation in severe ARDS, illustrated by a complicated case. AB - When conventional respiratory strategies fail to maintain adequate oxygenation treatment of severe ARDS is largely empirical. Modern techniques such as inverse ratio ventilation, permissive hypercapnia, NO inhalation and lowering tidal volumes/pressures are advocated. We report on a patient with severe ARDS who showed all the complications of the disease. The risks and benefits of (non)conventional ventilatory strategies are discussed and illustrated. PMID- 9852709 TI - Severe muscle weakness secondary to paraneoplastic hypophosphatemia in neuroblastoma. AB - We report a 28-year-old woman who presented with severe proximal muscle weakness secondary to paraneoplastic hypophosphatemia and associated with recurrent neuroblastoma. The biochemical findings included hyperphosphaturia, a reduced serum level of 1,25-dihydroxyvitamin-D3, elevated alkaline phosphatase and normocalcemia which are pathognomic for paraneoplastic hypophosphatemia. Following systemic chemotherapy and supplementation of 1,25-dihydroxyvitamin-D3 a complete remission of the neuroblastoma was achieved and all features of the paraneoplastic hypophosphatemia gradually disappeared. In the differential diagnosis of muscle weakness, hypophosphatemia should be included. Paraneoplastic hypophosphatemia associated with metastatic neuroblastoma has not been reported previously. Diagnosis, mechanism and therapy of paraneoplastic hypophosphatemia are shortly reviewed. PMID- 9852710 TI - ARDS, searching for treatment modes or understanding the "rules"? PMID- 9852711 TI - Community-acquired pneumonia "State-of-the-art". PMID- 9852712 TI - Effect of intravenous infusions on laboratory results in blood specimens drawn proximal to the insertion site of an intravenous canula. AB - BACKGROUND: Blood sampling contralateral to the insertion site of an intravenous line is always first choice. When blood must be drawn proximal to an i.v. insertion site, dilution and contamination by the infused substance can occur. To study the effects of obtaining blood specimens proximal to a discontinued intravenous line a clinical trial in a small community hospital was designed. METHODS: Ten patients received an intravenous infusion with sodium chloride 0.45%/dextrose 2.5%. Bloodsamples were taken simultaneously from the i.v.-arm and the control-arm with the infusion running and 1, 3, and 5 min after shutting off the infusion. RESULTS: Between-arm differences are significant for all analytes while the infusion is running, but not after the infusion is discontinued. Wide ranges in differences in terms of percentage exist for many analytes 1 min after shutting off the infusion, and for glucose also 3 min after shutting off the infusion. CONCLUSIONS: A three minute interval is recommended when drawing blood proximal to a shut-off infusion. Longer intervals may be advisable for analytes present in the infused substance. PMID- 9852713 TI - Tolerability and effects of high doses acipimox as additional lipid-lowering therapy in familial hypercholesterolemia. AB - BACKGROUND: Acipimox, a derivative of nicotinic acid, lowers serum lipid levels by reducing the production of very-low-density and low-density lipoproteins (LDL). METHODS: We studied the additional lipid-lowering effect of high doses of acipimox in 12 patients with severe familial hypercholesterolemia (FH) who were on treatment with an HMG CoA reductase inhibitor, in some cases in combination with a resin. RESULTS: There was a significant reduction in total serum cholesterol (-9%), LDL-cholesterol (-9%) and serum triglycerides (-21%) when the standard doses of acipimox (750 mg/day) was added to treatment with simvastatin (and a resin). However, higher doses had no further hypolipidemic effect. In concordance with the reduction of serum cholesterol and LDL-cholesterol there was a significant decrease in apolipoprotein (apo)-B (-11%). There was no change in HDL-cholesterol, apo-A1 and lipoprotein(a). Acipimox in high doses up to 2250 mg/d was well tolerated except for initial gastric complaints and of flushing; because of these side effects one patient dropped out of the study. CONCLUSIONS: Acipimox in high doses, which were well tolerated, has no additional lowering effect on LDL-levels compared to the standard dose in patients with severe FH who are already treated with simvastatin. PMID- 9852714 TI - [Developmental abnormalities of the eye]. PMID- 9852715 TI - [Effect of matrix metalloproteinase inhibitor on experimental pseudomonal corneal ulceration]. AB - We examined the effect of matrix metalloproteinase (MMP) inhibitor (CS-610) on experimental pseudomonal corneal ulceration by clinical and histological evaluation. Intrastromal injection of 3.5 microliters sterile culture broth of P. aeruginosa, IID-1117 (13.5 unit Type I collagenase equivalent proteinase activities), was done to induce corneal ulcers in guinea pigs. The animals were divided into two groups of 23 each. The CS-610 group received topical CS-610 (400 micrograms/ml) treatment at 2-hour intervals and the control group received only the vehicle of CS-610 at the same intervals. In the control group, corneas developed acute corneal damage following corneal ulcerations at 6-12 hours. In the CS-610 group, these corneal lesions were inhibited in most of the eyes (p < 0.01). In the late period, as inflammatory cells migrated into the cornea, some animals of the CS-610 group developed corneal ulcer. The results indicated that CS-610 had a potent inhibitory activity against pseudomonal proteinases in vivo. The results also suggested that the mechanism of the ulceration model involved not only pseudomonal proteinases but also endogenous responses. PMID- 9852716 TI - [Accumulation of a photosensitizer ATX-S 10 Na (II) to experimental corneal neovascularization]. AB - In order to determine the most appropriate time point for laser irradiation in photodynamic therapy with a new photosensitizer, ATX-S 10 Na (II), which produces selective occlusion of new vessels, we investigated the time course of plasma levels of ATX-S 10 Na (II) after intravenous administration and degree of dye accumulation in the corneal neovascularization in rabbit eyes. Plasma ATX-S 10 Na (II) concentration decreased rapidly after injection and become virtually undetectable at 24 h, indicating rapid excretion from the body. Nitrogen-pulsed laser spectrofluorometry demonstrated that the amount of ATX-S 10 Na (II) in new corneal vessels increased and reached a maximal level 2 to 4 hours after dye injection. Under a fluorescence microscope, ATX-S 10 Na (II) was localized in the wall new corneal vessels and in extravascular tissue 2 to 4 hours after dye injection. These results indicate that the appropriate time for laser irradiation in selective PDT is between 2 and 4 hours after dye injection, when a larger amount of dye is accumulated in neovasculature tissue compared to normal tissue. PMID- 9852717 TI - [Vascular endothelial growth factor levels in the aqueous and serum in diabetic retinopathy with or without neovascular glaucoma]. AB - We determined the levels of vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) in the aqueous and serum in non-insulin dependent diabetic patients with proliferative retinopathy (n = 12) and neovascular glaucoma (n = 11). The aqueous levels of PEGF/VPF were significantly higher in both groups than in 10 diabetics without such complications. The levels were very high in patients with neovascular glaucoma, suggesting that VEGF/VPF is involved in the pathogenesis and progression of diabetic neovascular glaucoma. The serum levels were not significantly related to the presence or the stage of retinopathy. The findings suggest the possibility of treatment of neovascular glaucoma using anti- VRGF/VFP preparations. PMID- 9852718 TI - [A familial case of proliferative diabetic retinopathy associated with a mutation in the mitochondrial gene]. AB - We report a 39-year-old male patient with proliferative diabetic retinopathy associated with a mutation in the mitochondrial DNA and with his family. The patient was referred for fundus examination in 1993. He had been diagnosed as having diabetes mellitus 15 years before but his diabetic control was not good. He was thin and short and presented with bilateral proliferative diabetic retinopathy. When he received pan retinal photocoagulation, an exudative retinal detachment was found in his left eye, but it disappeared spontaneously. The patient's history included sensorineural deafness and autonomic nerve system disorder. Mitochondrial analysis obtained from blood showed mutation of adenine to guanine at mitochondrial DNA 3243. Generally speaking, mitochondrial disorders often include pigmentary degeneration. For this reason diabetic patients with mitochondrial disorders rarely suffer proliferative diabetic retinopathy. In this case, the retinitis pigmentosa was not so severe as to cause the proliferative change, so that it was probably derived from heteroplasmy. PMID- 9852719 TI - [Glaucoma with microcornea; morphometry and differential diagnosis]. AB - To evaluate the characteristics of 10 glaucomatous cases with microcornea, we measured corneal diameter, curvature, axial length, and depth of anterior chamber, and examined the eyes with a specular microscope and an ultrasound biomicroscope. The ten cases examined in this study included 1 case of cornea plana, 2 cases of sclerocornea, 2 cases of nanophthalmos, and 5 cases of anterior microphthalmos. Three of the 10 cases were combined with open angle glaucoma, and the others with closed angle glaucoma. Open-angle glaucoma seemed like developmental glaucoma with angle maldevelopment. The closed angle type may appear at a younger age than in patients who have simple pupilary block. There are probably complex mechanisms related with closed angle glaucoma with microcornea. While differential diagnosis among these cases is relatively easy, we should evaluate how or why the intraocular pressure rises as far as possible before forming a treatment plan for each patient. PMID- 9852720 TI - [Relationship between the retinal nerve fiber layer thickness and the effect of aging in normal eyes]. AB - We studied the correlation between the thickness of the retinal nerve fiber layer and the role of age in 155 normal eyes of 83 subjects without significant ocular diseases. Subjects with a refractive error exceeding +/- 5 diopters were excluded. We divided the peripapillary area into four quadrants: superior (S), inferior (I), temporal (T), and nasal (N). We measured peripapillary retinal nerve fiber layer thickness (NFLT) in these four quadrants and in the total area (T(o)) using a confocal scanning laser polarimeter (Nerve Fiber Analyzer), and determined the correlation coefficients. The retinal nerve fiber layer thickness of the total area and of the four quadrants was not significantly associated with an increase in age. Since changes in the thickness were small in the nasal and temporal quadrants, we measured the relative ratios of the thickness in each quadrant to that in the nasal and temporal quadrants. The ratio of the total area to the nasal quadrant (T(o)/N), the superior to the nasal quadrant (S/N), the inferior to the nasal quadrant (I/N), the total area to the temporal quadrant (T(o)/T), the superior to the temporal quadrant (S/T), and the inferior to the temporal quadrant (I/T) was significantly associated with an increase in age. Multiple comparison in each age group revealed no age-related significant decreases in any of the parameters. These findings suggest that these parameters may reflect the degree of aging in normal eyes, but age-related abnormality in the retinal nerve fiber layer thickness may be small. PMID- 9852721 TI - [Ultrasound biomicroscopic findings of anterior hyaloidal fibrovascular proliferation]. AB - We performed ultrasound biomicroscopy (UBM) on 5 eyes with anterior hyaloidal fibrovascular proliferation (AHFVP), which had developed after vitrectomy for proliferative diabetic retinopathy. AHFVP was observed as a thick membrane or a mass image which extended from the vitreous base to the pars plicata, and was apparently differentiated from the anterior vitreous membrane and cilliary body. UBM showed proliferative stalks extending from sclerotomy sites toward the anterior vitreous membrane. These UBM images indicated that AHFVP originated from sclerotomy sites. UBM also enabled us to detect traction detachment of the peripheral retina associated with AHFVP, which could not be observed by conventional B-mode echography. Thus UBM is useful in establishing a diagnosis of AHFVP, and this technique is valuable to determine the indication and the timing of surgical treatment for AHFVP. PMID- 9852722 TI - [Meige's syndrome associated with basal ganglia and thalamic functional disorders]. AB - Magnetic resonance imaging (MRI) or single positron emission computed tomography (SPECT) or both were performed and the responses of surface electromyography (EMG) were examined in seven cases of Meige's syndrome. MRI or SPECT or both demonstrated lesions of the basal ganglia, the thalamus, or both in five of the cases. Surface EMG revealed abnormal burst discharges in the orbicularis oculi and a failure of reciprocal muscular activity between the frontalis and orbicularis oculi in all the cases. These findings suggest that voluntary motor control and reciprocal activity in the basal ganglia-thalamocortical circuits are impaired in Meige's syndrome. In addition, good responses were seen to clonazepam, tiapride and trihexyphenidyl in these cases. Therefore, we conclude that dopaminergic, cholinergic, and gamma-aminobutyric acid (GABA) ergic imbalances in the disorders of the basal ganglia and thalamus in Meige's syndrome cause control in the excitatory and inhibitory pathways to be lost, resulting in the failure of integration in reciprocal muscular activity and voluntary motor control. This failure subsequently causes the symptoms of Meige's syndrome. PMID- 9852723 TI - [Synthetic protease inhibitors for human acute pancreatitis]. PMID- 9852724 TI - [A randomized controlled study of total parenteral nutrition and enteral nutrition by elemental and polymeric diet as primary therapy in active phase of Crohn's disease]. AB - We performed randomized controlled study to compare the short-term therapeutic effect of total parenteral nutrition (TPN), elemental diet (ED) and polymeric diet (PD) given as primary therapy in active phase of Crohn's disease. In hospital for Crohn's disease, twenty-eight patients were given nutritional therapy: 9 patients by TPN, 10 by ED, and 9 by PD. Nutritional state, inflammatory reactions, disease activity and clinical remission rate were assessed two weeks and four weeks after treatment, and morphological findings were assessed before and after each nutritional therapy by radiographic and colonoscopic findings. Inflammatory reactions were more effectively controlled by TPN and ED than by PD, and early improvement achieved by TPN and ED was especially note-worthy. Clinical remission rate after treatment by TPN was highest in three types of nutritional approach, but no significant difference was seen at any point. In nutritional state, disease activity and morphological findings, comparable changes were effected without preference. These results suggest that nutritional therapy by total parenteral nutrition and elemental diet is superior to polymeric diet for treating active phase of Crohn's disease with marked inflammatory reactions. PMID- 9852725 TI - [Changes of 24-h Holter monitor recordings in association with interferon alpha therapy for chronic hepatitis C]. AB - We examined cardiovascular complication of interferon (IFN) therapy in 23 patients with chronic hepatitis C who did not have cardiac disease prospectively. Twenty four-h Holter monitor recordings were performed before and during IFN therapy. Seven of these patients (30%) showed abnormalities on their 24-h Holter monitoring recordings. Premature ventricular contraction (PVC) occurred in two patients, intermittent WPW syndrome in one, and ST-T change in four. Only one patient with PVC complained of palpitation. These complications were not severe and immediately after IFN therapy was stopped. There was no correlation between Holter ECG abnormalities and sex, age, quantity of HCV, or 2-5 oligoadenylate synthetase activity. It was suggested that cardiovascular complications caused by IFN therapy occurred more frequently than expected. However, diagnosis of these complications is difficult because most patients have no subjective symptoms and there is scarcely any change in laboratory test results. Careful observation of patients may be required during IFN therapy regardless of cardiovascular symptoms. PMID- 9852726 TI - [Continuous arterial infusion of protease inhibitor with supplementary therapy for the patients with severe acute pancreatitis--clinical effect of arterial injection of ulinastatin]. AB - We treated five patients with severe acute pancreatitis by continuous arterial infusion (CAI) of protease inhibitor, nafamostat mesilate. Arterial injection (AI) of ulinastatin was performed in four cases and AI of antibiotics (IPM/CS) was done in one case, as supplemental therapies of CAI. Abdominal pain disappeared in 7.9 hours on the average, abdominal tenderness disappeared in 5.0 days and laboratory data lately recovered. All five cases treated by these therapies were cured without hemodialysis or surgical treatment in acute phase. AI of ulinastatin through arterial infusion catheter is pharmacokinetically more effective, because it yields a relatively high concentration of the drug at the acting site when compared with that of intravenous injection. Furthermore ulinastatin inhibits different types of protease from nafamostat mesilate. Therefore the clinical effect of CAI of nafamostat mesilate is enhanced by the combined therapy with AI of ulinastatin. It is also suggested that arterial injection of ulinastatin might be effective for the control of abdominal pain and that arterial injection of antibiotics might have an advantage on prevention of infectious pancreatic necrosis. PMID- 9852727 TI - [Malignant gastrointestinal stromal tumor of the duodenum after gastrectomy due to early gastric cancer. Report of a case]. PMID- 9852728 TI - [A case report of intussusception due to an inflammatory fibroid polyp in the ileum]. PMID- 9852729 TI - [A case of Cowden's disease associated with rectal carcinoid]. PMID- 9852730 TI - [A report of a case with collagenous ileocolitis]. PMID- 9852731 TI - [A case of primary hepatic lymphoma]. PMID- 9852732 TI - [An adult case of extra-hepatic portal obstruction with bleeding from esophageal varices]. PMID- 9852733 TI - [A case of idiopathic iliopsoas hematoma associated with liver cirrhosis]. PMID- 9852734 TI - [Biliobiliary fistula (Corlette type I): a case report with preoperative diagnosis by magnetic resonance cholangio-pancreatography]. PMID- 9852735 TI - [Two cases of anorectal Crohn's disease with characteristic appearances of anus]. PMID- 9852736 TI - Mechanical properties of vertebrate hard tissues. PMID- 9852737 TI - Collagen--calcium phosphate composites. AB - There is a widespread clinical need for bone augmentation and replacement. The major solid phases of bone are collagen and calcium phosphate and a bone analogue based on these two constituents should have some useful properties. In this review this theme is developed and the properties of natural and naturally based composites are compared. Composites have been produced by the precipitation of calcium phosphates on to collagen and a summary of the methods and results from mechanical testing and scanning electron microscopy are presented. Composites with mechanical properties intermediate between cancellous and cortical bone have been produced. The review concludes by explaining some of the mechanical properties of the composites, using knowledge of the hierarchical architecture of bone and results from microscopical examination of the fractured composites. PMID- 9852738 TI - Osteotransductive bone cements. AB - Calcium phosphate bone cements (CPBCs) are osteotransductive, i.e. after implantation in bone they are transformed into new bone tissue. Furthermore, due to the fact that they are mouldable, their osteointegration is immediate. Their chemistry has been established previously. Some CPBCs contain amorphous calcium phosphate (ACP) and set by a sol-gel transition. The others are crystalline and can give as the reaction product dicalcium phosphate dihydrate (DCPD), calcium deficient hydroxyapatite (CDHA), carbonated apatite (CA) or hydroxyapatite (HA). Mixed-type gypsum-DCPD cements are also described. In vivo rates of osteotransduction vary as follows: gypsum-DCPD > DCPD > CDHA approximately CA > HA. The osteotransduction of CDHA-type cements may be increased by adding dicalcium phosphate anhydrous (DCP) and/or CaCO3 to the cement powder. CPBCs can be used for healing of bone defects, bone augmentation and bone reconstruction. Incorporation of drugs like antibiotics and bone morphogenetic protein is envisaged. Load-bearing applications are allowed for CHDA-type, CA-type and HA type CPBCs as they have a higher compressive strength than human trabecular bone (10 MPa). PMID- 9852739 TI - Histomorphological and biomechanical characterization of calcium phosphates in the osseous environment. AB - The standardization of characterization techniques is becoming increasingly important for bone replacement materials as it becomes apparent that, for the field to advance, testing must be developed to allow the biocompatibility or bioactivity of a new material to be assessed and directly compared with existing materials. Currently there are many forms of biocompatibility test for materials destined for the osseous environment, ranging from immersion in simulated body fluid to implantation into living bone. However, the variety of ways in which the data from these tests may be acquired and interpreted, as a result of changes in parameters such as surgical technique and mechanical test conditions, means that much of the published data within the field is not comparable. This paper will introduce the concept of biocompatibility by considering calcium phosphate bioceramics, and discusses some aspects of in vivo experimental design, including simple histomorphometry techniques, in addition to considering practical methods for the assessment of the biomechanical characteristics of an osseointegrated implant. PMID- 9852740 TI - Polymers in dentistry. AB - There is a wide choice of materials available for restorative dentistry covering a range of requirements. Fundamental knowledge about the properties of the polymers in use in dentistry is an advantage as it provides information relevant to clinical practice. Dentistry, perhaps, has the unique distinction of using the widest variety of materials, ranging from polymers, metal and metal alloys, ceramics, inorganic salts and composite materials. In the present paper, polymers and polymer composites used directly or indirectly for restorations, prostheses or for production of appliances in dentistry is discussed. PMID- 9852741 TI - Biomaterials and cardiovascular devices. AB - In the field of cardiovascular surgery there is presently a lack of biomaterials possessing essential characteristics of the native tissue or organ which is to be replaced. This paper describes various biomaterials that have been introduced into the circulatory system and the complex reactions that subsequently occur. The risk of infection is also discussed as well as prevention and treatment regimes that can be used. Examples of future biomaterial development are outlined in an attempt to achieve biocompatibility. PMID- 9852742 TI - Shape memory alloys for medical applications. AB - The shape memory alloys exhibit a number of remarkable properties, which open new possibilities in engineering and more specifically in biomedical engineering. The most important alloy used in biomedical applications is NiTi. This alloy combines the characteristics of the shape memory effect and superelasticity with excellent corrosion resistance, wear characteristics, mechanical properties and a good biocompatibility. These properties make it an ideal biological engineering material, especially in orthopaedic surgery and orthodontics. In this work the basis of the memory effect lies in the fact that the materials exhibiting such a property undergo a thermoelastic martensitic transformation. In order to understand even the most elementary engineering aspects of the shape memory effect it is necessary to review some basic principles of the formation and the characteristics of the martensitic phase. The different properties of shape memory, superelasticity, two-way shape memory, rubber-like behaviour and a high damping capacity are reviewed. Some applications proposed in recent years are described and classified according to different medical fields. PMID- 9852743 TI - Applications of computer modelling for the design of orthopaedic, dental and cardiovascular biomaterials. AB - Biomaterials do not escape from the general trend present in all contemporary science and technology towards increasing use of computers and information technology. In this paper the use of computer modelling for the design of biomaterials is discussed. The word 'biomaterials' is interpreted in its broadest sense, i.e. referring to any foreign object brought into the body for temporary or permanent use. Computer modelling will first be discussed as a tool to model biological structures (bones, arteries) or to investigate and simulate biological interactions at implant-host interfaces. It will then be illustrated how computer modelling, using insights gained from the modelling of the biological structures themselves, is used in the design process of dental, orthopaedic and cardiovascular prostheses. The area of computer modelling for biomaterials applications has become so vast that an exhaustive overview is impossible in the framework of one paper. Rather, some illustrative case studies will be discussed which are, in the opinion of the authors, representative of general trends in this challenging domain of science on the boundary between engineering and medicine. PMID- 9852744 TI - [A viewpoint: overlap syndrome in diffuse collagen diseases]. PMID- 9852745 TI - [Diagnosis for human parvovirus B19-polyarthritis: usefulness of empty particle B19.ELISA and B19-DNA.PCR]. AB - OBJECTIVE: To evaluate the usefulness of new ELISA for human parvovirus B19 (B19) antibodies and PCR for the diagnosis of acute onset of B19 polyarthritis. METHODS: We evaluated the reproducibility and sensitivity on the detection of anti-B19 antibody by ELISA using recombinant VP-1 and VP-2 (empty particle), and then studied for the prevalence of IgM and IgG B19 antibody in 125 samples for anti-B19 tests. The random study on anti-B19 antibody assay as well as PCR for B19-DNA was also performed in 130 cases with acute onset of arthritis excluding those with known origins, 224 with rheumatoid arthritis and 149 with other categories. RESULTS: The results by using B19-empty particle ELISA were reproducible and showed the assay was a sensitive way for clinical use. IgM anti B19 antibodies were positive not only in all samples from erythema infectiosum, but also often in those from hemolytic anemia, pure red cell aplasia, fetal hydrops, hepatic injury, fever of unknown origin. Among 130 with acute onset of arthritis, 21 showed positive tests for IgM anti-B19 antibody and/or B19 DNA. On the other hand, 4 among 224 patients with rheumatoid arthritis were positive for IgM anti-B19 antibody, but all of 149 in control group were negative for IgM anti B19 antibodies and for B19 DNA. CONCLUSION AND DISCUSSION: Anti-B19 ELISA using B19-empty particle which has been introduced as a routine test system, is a useful tool for the diagnosis of acute onset of B19 arthritis. An additional examination using PCR for B19 DNA may contribute for understanding persistent B19 polyarthritis or reactivation of B19 infection. PMID- 9852746 TI - [Clinical study on five myeloperoxidase specific anti-neutrophil cytoplasmic antibody (MPO-ANCA) positive Churg-Strauss syndrome cases]. AB - OBJECTIVE: In order to clarify the clinical characteristics of ANCA positive Churg-Strauss syndrome (CSS), ten CSS patients were analyzed. METHODS: ANCA was detected using an immunofluorescence (IF) assay and ELISA. ANCA was measured in both active and inactive stage of the disease. RESULTS: ANCA was present in 5 patients (5/10 : 50%) with active CSS phase and all ANCA patient results showed perinuclear pattern by IF assay and myeloperoxidase specific results by ELISA. Patients with ANCA were older (average age : 58.0) and had higher incidence of renal involvement (4/5 : 80%) than was found with ANCA negative patients (average age; 38.4, no patients with renal involvements). Among 5 patients with ANCA, one had rapidly progressive glomerulonephritis and one had alveolar hemorrhage. Remission was induced for 4 ANCA positive patients by corticosteroid and/or immunosuppressive drug and one required long term maintenance hemodialysis. ANCA titer changed in parallel with disease activity, such as proteinuria, hematuria and alveolar hemorrhage. No significant differences in regard to eosinophilia, serum IgE and CRP titer were found between ANCA positive and negative patients. CONCLUSION: These results strongly indicate that MPO-ANCA may play an important role in the pathogenesis of vascultis, such as renal injury and alveolar hemorrhage in CSS. PMID- 9852747 TI - [Plain film and CT findings of pulmonary involvement in rheumatoid arthritis]. AB - OBJECTIVES: To analyze the plain film and CT findings of rheumatoid arthritis (RA)-associated lung diseases and to correlate them with clinical manifestations. METHODS/MATERIALS: We retrospectively reviewed the plain chest and CT findings of 51 RA patients with lung diseases. Sixteen CT findings were separately corded as present or absent, and drew up our classification of lung diseases as follows; 1. interstitial pneumonia/pulmonary fibrosis (IP/PF), 2. airway disease (AD), 3. parenchymal disease (PD), and 4. mixed. Four plain film findings were also recorded, and classified in IP/PF or AD group. This classification was analyzed with reference to clinical parameters and courses. RESULTS: Fifty-one patients were classified as follows; 1. IP/PF (n = 32, 62.6%), 2. AD (n = 15, 29.4%), 3. PD (n = 3, 5.9%), 4. mixed (n = 1, 2.0%). CT findings of PD were compatible with bronchiolitis obliterans organizing pneumonia with rapid improvement. IP/PF diagnosed with CT was depicted by the plain chest in 25 patients (78.1%). There is no false positive of the plain chest diagnosis in terms of IP/PF. As compared with IP/PF, AD consisted of significantly higher prevalence in female patients and patients with severe articular involvement and sinusitis. In spite of variable outcome in patients with AD, those with IP/PF showed the progression of honeycombing, and 4 of them were died from respiratory failure. CONCLUSIONS: CT based classification of rheumatoid lung diseases correlates with clinical manifestations and prognosis and is useful for clinical management in patients of RA. CT diagnosis is useful especially when plain chest does not demonstrate typical IP/PF pattern. PMID- 9852748 TI - [Three cases of relapsing polychondritis--triggering factor and treatment of relapses]. AB - We present three cases of relapsing polychondritis (RP), all of which were confirmed by biopsy. In the first case, where the symptoms appeared after treatment for kidney and lung cancers, it was thought that interferon alpha, radiation or photodynamic therapy had been the causative or triggering factor. In the second case, eye surgery may have triggered the recurrence of RP. The possibility that medical intervention might cause, promote or trigger RP by disturbing the immune system was discussed. In the third case, where the chief complaint was dyspnea due to severe bronchial constriction, the importance of assessing the state of the respiratory tract, for instance by spiral 3 D CT imaging of the chest, was suggested. The appropriate use of tracheobronchial stents for some cases associated with dyspnea was also discussed. PMID- 9852749 TI - [A case developing tetraplegia due to systemic lupus erythematosus which was remitted by a steroid]. AB - The case reported here was a 58-year-old woman who was diagnosed as having systemic lupus erythematosus (SLE) in 1985 because she had erythema in the cheeks arthritis, a hematological abnormality (decreased white blood cell count), an immunological abnormality (LE-positive cells), and a positive result of antinuclear antibody test. Although the patient was once remitted by treatment with prednisolone (PSL) at 60 mg/day, and continuously received PSL at a maintenance dose of 2.5 mg/day, she was admitted in June 1996 by our hospital with chief complaints of fever and decreased muscular strength in the four extremities. At admission, she had symmetrical tetraplegia, which was peripherally predominant and severer in the lower extremities, and hypoesthesia accompanied by numbness. She was negative for anti-phospholipid antibody and showed no abnormality in cerebrospinal fluid examination. No lesions responsible for tetraplegia were detected at brain MRI, spinal MRI, or myelography. Because fever, multiple arthralgia, an increased erythrocyte sedimentation rate, a decreased lymphocyte count, hypocomplementemia, and a high immune complex level indicated the active stage of SLE (recurrence), she was given PSL at dose increased to 60 mg/day. After about 2 months, SLE was remitted and her tetraplegia and hypoesthesia was gradually improved thereafter. Although tetraplegia associated with peripheral nervous disorder, in which angitis occurring as a symptom of the active stage of SLE was thought to be Involved, was remitted by a steroid in out patient, no such cases have been reported in Japan, to our knowledge. Thus our patient was thought to be a very rare case of value. PMID- 9852750 TI - [Hemophagocytic syndrome in a patient with rheumatoid arthritis]. AB - A 62-year-old man with 16 year-history of rheumatoid arthritis (RA) was admitted due to progressive pancytopenia, general fatigue, and high fever. He was treated with 5 mg methotrexate weekly in RA. His bone marrow examination revealed a decreased nuclear cell count (2.1 x 10(4)/microliter), megakaryocyte count (16/microliter), and macrophages phagocytizing blood cells (13.2%), indicating the presence of hemophagocytic syndrome. No infections agent was detected in cultures derived from his blood or other sources. The serological tests for several viruses revealed no obvious viral etiology. The systemic lymphonodes were not swelling. Administration of 40 mg prednisolone daily improved his abnormal hematological findings. This is a case of RA accompanied by hemophagocytic syndrome, which is a rare complication of RA. PMID- 9852751 TI - [A case report of selective IgA deficiency in rheumatoid arthritis and anti-IgA antibody induced anaphylactic transfusion]. AB - We described a case of rheumatoid arthritis (RA) with selective IgA deficiency. A 69 year-old female with RA was admitted because of gall bladder cancer, and also had selective IgA deficiency which serum IgA level was less than 5.0 mg/dl, and IgA 1 and IgA 2 subclasses were not detected. Prior to the operation, she was given red cell compatible blood transfusion because of severe anemia. After 30 min of transfusion, she developed chill, nausea, vomiting and hypotension. These anaphylactic reactions might be induced by the presence of anti-IgA antibody, since the level of this antibody titers in her serum was elevated, assessed by the methods of ELISA and Western blotting. Although a case of RA associated with selective IgA deficiency, and also with elevated serum anti-IgA antibody level is extremely uncommon, attention should be paid to the presence of anti-IgA antibody in patients with selective IgA deficiency to avoid any unexpected anaphylactic reactions. PMID- 9852752 TI - [A case with rheumatoid arthritis complicated with ANCA-associated vasculitis]. AB - A 50-year-old woman with rheumatoid arthritis (RA) developed constitutive symptoms such as fever and weight loss, mononeuritis multiplex and rapidly progressive glomerulonephritis (RPGN). The renal biopsy revealed crescentic glomerulonephritis (CrGN) with few immune deposits such as IgG and C 3 and necrotizing vasculitis, which led to the pathological diagnosis of microscopic polyangiitis (MPA). Moreover the high serum level of anti-myeloperoxidase (MPO) antibody, one of the anti-neutrophil cytoplasmic antibody (ANCA), suggested that she had ANCA-associated vasculitis. The renal prognosis of the ANCA-associated vasculitis is said to be poor unless patients were treated in the early phase of the disease. As we started to treat the patient when her serum creatinine level (sCr) was 1.7 mg/dl and creatinine clearance (CCr) was 27 ml/min, her renal function returned to the almost normal level (sCr = 0.6, CCr = 91). It is well known that patients with RA may develop various kinds of extraarticular manifestations which are usually related to immune-complex mediated vasculitis, what we call malignant RA. We emphasize that ANCA-associated vasculitis is another important complication of RA. When we see a RA patient with constitutional symptom, abnormal urinary sediments and other clinical signs of vasculitis such as mononeuritis multiplex, ANCA-associated vasculitis should be considered. Since the early diagnosis and treatment of ANCA-associated vasculitis is a key to prevent renal failure, it is encouraged to measure the serum ANCA titer for not only the diagnosis of such patients but the evaluation of their clinical course. PMID- 9852753 TI - [Possible pathogenic role of IL-16 on SLE: with reference to HIV-1 infection]. PMID- 9852754 TI - [Diagnosis and management of early rheumatoid arthritis]. PMID- 9852755 TI - Cytoplasmic regulatory functions of the KH-domain proteins hnRNPs K and E1/E2. PMID- 9852756 TI - Does NADH play a central role in energy metabolism in Helicobacter pylori? AB - The complete genome sequence of Helicobacter pylori reveals an unusual NADH quinone oxidoreductase (NDH-1 or Complex I) that might lack the NADH-binding domain. H. pylori also lacks various NADH-generating enzymes. What are the consequences for electron transfer to H. pylori NDH-1 and could NADPH be involved? PMID- 9852757 TI - Cofactor- and substrate-binding domains in flavin-dependent N-hydroxylating enzymes. PMID- 9852758 TI - The frizzled motif: in how many different protein families does it occur? PMID- 9852759 TI - Agonist-independent regulation of constitutively active G-protein-coupled receptors. AB - G-protein-coupled receptors constitute one of the largest protein super-families in mammals. Since the cloning of the encoding genes, these important drug targets have been subjected to thorough biochemical and pharmacological studies. It has become clear that G-protein-coupled receptors not only transmit signals after stimulation by agonists but can also spontaneously couple to signal-transduction pathways. Recent findings show that constitutively active G-protein-coupled receptors can also be regulated in an agonist-independent manner, which has important implications for the interpretation of the actions of (inverse) agonists and the results of site-directed-mutagenesis studies. PMID- 9852760 TI - Eukaryotic lipid-biosynthetic enzymes: the same but not the same. AB - The discovery that different eukaryotic enzymes catalyse the same lipid biosynthetic reaction has paralleled the recognition that segregated pools of lipids have unique biological functions. This review considers enzymes involved in the synthesis of diverse classes of lipids--glycerolipid precursors, phospholipids, sterols and eicosanoids--and summarizes recent data that show that these duplicate enzymes are frequently encoded by different genes and have unique subcellular locations. Does this duality merely represent redundancy or do the different isoforms provide pools of lipids for specific biological purposes? PMID- 9852762 TI - Contrasting lifestyles of rolling-circle phages and plasmids. AB - The rolling-circle mechanism of DNA replication is used by small prokaryotic genomes, such as single-stranded phages and plasmids. However, phages and plasmids have adapted the rolling-circle mechanism differently to suit their contrasting biological needs. The phi X174 phage uses a monomeric initiator protein catalytically, displays incomplete termination and recycles the initiator protein, in order to mass-produce phage progeny. By contrast, to control replication precisely, the pT181 plasmid uses a dimeric initiator protein stochiometrically, completes termination and inactivates the initiator after each replication cycle. The phi X174 phage and the pT181 plasmid represent paradigmatic adaptations of the rolling-circle mechanism and could provide models for other replicons. PMID- 9852761 TI - The structural and mechanochemical cycle of kinesin. AB - Kinesin is a microtubule-based motor protein that pulls vesicles or organelles towards the plus end of microtubules. Structural changes in the protein that drive motility are coupled to ATP binding and hydrolysis. Here, we attempt to integrate recent structural and kinetic results into a picture of the mechanochemical cycle of kinesin. PMID- 9852763 TI - Harnessing free radicals: formation and function of the tyrosyl radical in ribonucleotide reductase. AB - Ribonucleotide reductases (RNRs) are uniquely responsible for converting nucleotides to deoxynucleotides in all organisms. The cofactor of class-I RNRs comprises a di-iron cluster and a tyrosyl radical, and is essential for initiation of radical-dependent nucleotide reduction. Recently, much progress has been made in understanding the mechanism by which this cofactor is generated in vitro and in vivo, as well as the function of the tyrosyl radical in nucleotide reduction. The Escherichia coli RNR cofactor provides a paradigm for cofactors in other di-iron requiring or tyrosyl-radical-requiring proteins. PMID- 9852764 TI - Iterated profile searches with PSI-BLAST--a tool for discovery in protein databases. PMID- 9852765 TI - Eukaryotic rRNA methylation: the calm before the Sno storm. PMID- 9852766 TI - Biochemical and hormonal changes associated with experimental infection of chicks with infectious bursal disease virus. AB - The inoculation of chicks with the infectious bursal disease (IBD) virus manifested typical clinical signs indicative of IBD viral infection. The inoculated birds seroconverted and showed significantly decreased total protein, lipid and a decrease in the albumin to globulin ratio. A significant increase was seen in the concentration of corticosterone and thyroxine but not in the triiodothyronine level. PMID- 9852767 TI - A case control study of potential enteric pathogens for calves raised in cow-calf herds. AB - A matched case control study was performed to describe the epidemiological features of potential enteric pathogens for calves reared in 53 cow-calf herds located in western Switzerland. A total of 106 diarrhoeic calves and 126 healthy control calves were collected, all calves were less than 4 months old. Faecal samples were analysed for presence of infectious agents related to calf diarrhoea including enterotoxigenic E. coli, Verotoxin producing E. coli (VTEC), Campylobacter sp., Yersinia sp., Salmonella sp., rotavirus, coronavirus, helminths and coccidian protozoa. Multivariate logistic models were used to analyse the relationship between presence of infection and onset of diarrhoea. The study provided evidence of significant associations between diarrhoea and infection with rotavirus, Campylobacter coli and the presence of Verotoxin in faecal samples. With the exception of Cryptosporidium parvum intestinal parasites including Strongylidae and Eimeria sp. were found to be less prevalent in cases than in controls. Control calves were significantly more frequently infected with Strongyloides papillosus than case animals. PMID- 9852768 TI - Apoptosis of lymphocytes in mice administered lipopolysaccharide from Leptospira interrogans. AB - We report that administration of leptospiral lipopolysaccharide (LPS) in mice results in massive surface marker changes in the lymphocytes of the spleen. It appears that many of these changes relate to the large number of cells undergoing apoptosis. It is also shown that tumor necrosis factor-alpha (TNF-alpha) induces similar effects and is produced in large quantities after injection of leptospiral LPS. It is likely that cytokines such as TNF-alpha are involved in apoptosis. PMID- 9852769 TI - Decreased growth of Streptococcus uberis in milk from mammary glands of cows challenged with the same mastitis pathogen. AB - Milk samples from mammary glands challenged with Streptococcus uberis and from unchallenged mammary glands were selected for analyses of bacterial growth, antibody response, and lactoperoxidase activity. All challenged mammary glands became infected with isolation of S. uberis and elevated somatic cell counts in milk during the first week after challenge. In vitro growth of the homologous challenge strain and a heterologous strain of S. uberis was significantly lower in milk from challenged mammary glands than in milk from control mammary glands at 3, 5, and 7 days after challenge. Removal of casein significantly reduced bacterial growth. In general, antibodies specific to S. uberis started to increase at day 3 post-challenge and were higher in milk from challenged mammary glands than in milk from control mammary glands. There was also a marked increase in total IgG in milk from challenged mammary glands. Growth of S. uberis increased following heat treatment at 56 degrees C of pooled milk or whey samples from challenged mammary glands. Growth of S. uberis correlated negatively with the specific antibody response to the bacteria (P < 0.001). Lactoperoxidase activity varied among cows and among different samples over time and did not appear to contribute to decreased growth of S. uberis. These results suggest that decreased growth of S. uberis in milk from challenged mammary glands in comparison to milk from control mammary glands could result from the interaction of antibodies with complement components. PMID- 9852770 TI - [Model investigations of the impedance effectiveness conerning bacterial relevant to food hygiene]. AB - The impedance technique mostly meets today's requirements of microbiological rapid methods. At relatively high prime cost for the equipment the advantages are marked by low personnel and material costs as well as swiftness combined with highly flexible usage. The method is applicable for both quantitative and qualitative examinations but can fail occasionally in total count determination, especially if the sample material contains heterogeneous microbes. In model investigations with 53 strains of 17 different genera Enterobacteriaceae strains, Aeromonads and Enterococcus strains proved to be highly impedance effective. Lactobacillus strains and Pseudomonads as well as Staphylococcus aureus strains showed a low impedance effectiveness. Several strains, for example of the genera Micrococcus, Acinetobacter and Brochothrix, did not show any changes of the medium impedance under the chosen conditions. Criterion for characterization of impedance effectiveness was the impedance detection time starting with identical initial counts (10(3) cfu/ml). Impedance effectiveness of microbes was determined at highly varying degree by the parameters of generation time, lag-phase duration and relative activity. This can lead either to wrong negative (underestimations) or wrong positive (overestimations) results of bacterial count. PMID- 9852771 TI - Properties of serological group B streptococci of dog, cat and monkey origin. AB - This study was designed to identify and characterize further Streptococcus agalactiae isolated during routine diagnostics from three diseased dogs and a cat, as well as from the inner organs of a monkey which died on a sepsis with beta-haemolytic streptococci. The cultures could be identified as streptococci of serological group B by cultural, biochemical and serological properties and by restriction fragment length polymorphism analysis of polymerase chain reaction (PCR)-amplified 16S ribosomal DNA. A further characterization of the isolates by serotyping and by determination of antibiotic resistances revealed a close relationship of these isolates to the human biotype of this species. PMID- 9852772 TI - Haemonchus contortus: characterization and purification of excretory/secretory protease(s). AB - The activity of protease(s) has been examined separately in excretory/secretory (E/S) products from male and female Haemonchus contortus (Nematoda: Trichostrongylidae). The E/S proteolytic activity indicated the presence of preabsorptive digestion of host blood and/or tissues. Protease activity was optimum at 37 degrees C, pH 8.5 and 8.0 mg casein. These protease(s) were purified to 32.16- and 88.80-folds from male and female E/S products, respectively, by sequential purification with saturated ammonium sulphate followed by ion-exchange chromatography. The purification study revealed the presence of isomeric forms of protease(s) in the E/S products of H. contortus. PMID- 9852773 TI - Erysipelothrix septicaemia in neonatal lambs. PMID- 9852775 TI - Toxicants in the environment: are we being exposed? PMID- 9852776 TI - A microscale electrospray interface incorporating a monolithic, poly(styrene divinylbenzene) support for on-line liquid chromatography/tandem mass spectrometry analysis of peptides and proteins. AB - A methodology is described for creating a monolithic chromatography support within a pulled fused-silica electrospray needle. The monolith was formed from a mixture of styrene, divinylbenzene, 1-dodecanol, and toluene using 2,2' azobis(isobutyronitrile) as the catalyst. The mixture was loaded into 150-micron i.d. fused-silica capillary tubing with a pulled 5-10-micron needle tip at one end. Polymerization at 65 degrees C followed by removal of the porogen material yielded a stable, porous, monolithic support which had excellent properties for the separation and on-line, electrospray, mass spectrometry analysis of peptides and proteins. The performance of the monolith-filled electrospray needles was compared with similar needles filled with commercial C18 silica and polymeric particulate supports. Separation efficiencies for both protein and peptide mixtures were generally equal to or better than the particulate supports at comparable pressures and flow rates. The ion chromatograms derived from the on line MS analysis were remarkably free from chemical background signals that often complicate the LC/MS analysis of femtomole amounts of sample. Good sequence coverage was obtained by LC/MS/MS analysis of the peptide mixture obtained from a protein isolated by silver-stained gel electrophoresis. The capability of the monolith to do peak parking experiments was demonstrated by the characterization of an immunoreactive HPLC fraction. The simple fabrication method, chromatographic performance, and robust nature of these microscale integrated column electrospray sources make them ideally suited for high-sensitivity tandem LC/MS analyses. PMID- 9852777 TI - Applications of 1.06-micron IR laser desorption on a Fourier transform mass spectrometer. AB - Various sugars, peptides, and lipids were analyzed on a Fourier transform mass spectrometer using laser desorption and ionization with and without the assistance of matrixes. A compact Nd:YAG laser with an output at 1.06 microns corresponding to fundamental frequency was employed. Gram-negative and Gram positive bacteria were also subjected to laser desorption mass spectrometry. Characteristics ions of conjugated lipid, formed by attachment of alkali metal cations, endogenous to the cells, were observed. Particle/liquid matrixes (e.g., cobalt in glycerol) proved to be useful with the 1.06-micron laser. The particles absorb efficiently laser radiation in a broad wavelength range. The liquid provides the same advantages as in fast atom bombardment: increased signal-to noise ratios and enhanced sample lifetimes. The effect of laser power on total ion current was shown to differ for samples with and without the particle/liquid matrix. The Fourier transform analyzer provides MS/MS capability for both positive and negative ions from complex mixtures. Ions desorbed externally are introduced into the cell via a quadrupole ion guide with a lower mass cutoff. Such a setup allows matrix ions to be excluded and thus provides excellent signal to-noise ratios for lower mass range fragment ions formed inside the cell. PMID- 9852778 TI - Development and cellular applications of fiber optic nitric oxide sensors based on a gold-adsorbed fluorophore. AB - A new design for optochemical sensors has been applied to the development of a nitric oxide selective fiber optic sensor. This sensor is composed of a fluorescein derivative dye attached to colloidal gold. The fluorescein dye rearranges as nitric oxide adsorbs onto the gold, inducing a decrease in the fluorescence intensity of the dye. This mechanism has allowed preparation of fiber optic dye-based nitric oxide sensors, which have been made ratio-metric by addition of reference dye microspheres. Previously developed fast, selective optical sensors for detection of aqueous nitric oxide involved a protein, such as cytochrome c'. The new fluorescein derivative chemical sensors have characteristics similar to those of the protein-based biosensors, including fast response times, excellent selectivity, and complete reversibility. In addition, the chemical sensors have advantages such as greater stability and commercially available components. These sensors were utilized to measure nitric oxide production by BALB/c mouse macrophages. PMID- 9852779 TI - Analysis of phospho- and glycopolypeptides with infrared matrix-assisted laser desorption and ionization. AB - The analytical characteristics of infrared (IR) matrix-assisted laser desorption and ionization (MALDI) were investigated for the analysis of phosphopeptides, a phosphopolypeptide, and glycopeptides. Two commercially available instruments, a high-resolution delayed extraction (DE) reflectron time-of-flight (RETOF) mass spectrometer and a high-power pulsed Er:YAG laser, were interfaced to produce a high-resolution MALDI-DE-RETOF instrument that is easy to use and can be switched between UV- and IR-MALDI mode within seconds. In the interface design, particular attention was paid to maintaining the same professional operating environment for the new IR-MALDI mode as exists for the commercial UV-MALDI mode. This instrument configuration facilitates comparative observation and investigation of the relative analytical merits of IR- and UV-MALDI. The results of studies of the tryptic alpha-casein phosphopeptides, RP1 (a Thr45-monophosphorylated congener of the recombinant protein hirudin variant 1), and fetuin Asn81 tryptic glycopeptides are presented. The elimination of labile substituents such as phosphoric acid and sialic acid is suppressed in IR-MALDI-RETOF mass spectrometry, with concomitant higher analyte ion yields. These results reflect the advantages that accrue from deposition of significantly less internal energy in the case of IR-MALDI. PMID- 9852780 TI - Characterization of microdialysis acidification for capillary isoelectric focusing-microelectrospray ionization mass spectrometry. AB - A microdialysis junction, based on a microdialysis membrane connecting a separation capillary and a short, sharply tapered microelectrospray emitter capillary, is demonstrated for on-line combination of capillary isoelectric focusing (CIEF) with electrospray ionization mass spectrometry (ESI-MS). The microdialysis junction provides the necessary electrical connection across the dialysis membrane for defining the electric fields needed for the CIEF separation and the electrospray process. Additionally, postseparation acidification of focused protein zones eluted from the CIEF capillary is achieved using the microdialysis junction while separation efficiency and resolution is maintained. A microelectrospray emitter produces a stable electrospray of protein analytes without the need for a makeup liquid flow and eliminates any subsequent sample dilution and reduction in MS sensitivity. The microdialysis junction is advantageous over the coaxial liquid sheath interface as evidenced by the simplicity in operation procedures, the enhancement in detection sensitivity, and the linear correlation between protein migration time and isoelectric point in CIEF-ESI-MS. PMID- 9852781 TI - Structural assignment of permethylated oligosaccharide subunits using sequential tandem mass spectrometry. AB - The sequential tandem mass spectrometry (MSn) capabilities offered by quadrupole ion trap instruments have been explored in a systematic study of permethylated oligosaccharides. Under collision-induced dissociation, protonated molecular species generated in the electrospray ionization mode yield simple and predictable mass spectra. Information on sequence, branching, and, to some extent, interglycosidic linkages can be deduced from fragments resulting from the cleavage of glycosidic bonds. Simple rules for the structural assignment of carbohydrates have been established for the fragmentation of protonated species and subunits thereof and corroborated by 18O-labeling experiments. Moreover, sequential tandem mass spectrometry was demonstrated to allow the straightforward structural characterization of unknown carbohydrate moieties by comparing their CID spectra with those of a set of references. As the collision-induced dissociation patterns are not dependent on the number of prior tandem mass spectrometric steps, structures can be unambiguously assigned by match of the spectra. These findings establish the basis of MSn performed on a quadrupole ion trap instrument for elucidating structures of large carbohydrates, which can be virtually degraded in the mass spectrometer into smaller entities in one or several steps. This powerful technique has been applied, used in conjunction with specific CD3 labeling, to the characterization of series of subunits generated from fucosylated and sialylated oligosaccharides, which are among the most important structures as far as biological activities are concerned. PMID- 9852782 TI - Isolation of priority polycyclic aromatic hydrocarbons from natural sediments and sludge reference materials by an anti-fluorene immunosorbent followed by liquid chromatography and diode array detection. AB - The selective isolation of PAHs from complex environmental mixtures was accomplished by means of a new methodology based on antigen--antibody interactions. This method consists on the extraction of PAHs from water samples onto an anti-fluorene immunosorbent (IS) followed by liquid chromatography with diode array detection. Environmental sediments and a sludge reference material containing PAHs were analyzed using this methodology in order to validate the performance of the IS for the cleanup procedure of such materials. Sediments were extracted by sonication with dichloromethane/methanol (2:1), and the extracts were brought to a volume of 100 mL of water in order to perform the extraction with the anti-fluorene IS. The reliability of the cleanup achieved by the IS was well demonstrated in the analysis of sediment and sludge complex samples containing the priority PAHs established by the U.S. EPA at concentrations varying from 56 micrograms/kg to 26 mg/kg. Results were compared to those obtained with conventional cleanup procedures showing a better selectivity for PAHs. The chromatograms presented a clear baseline allowing the determination and quantification of PAHs at the ppb level. Using immunosorbents, extraction, trace enrichment, and cleanup were accomplished in only one step. PMID- 9852783 TI - Determination of C-21 ketosteroids in serum using trifluoromethanesulfonic acid catalyzed precolumn dansylation and 1,1'-oxalyldiimidazole postcolumn peroxyoxalate chemiluminescence detection. AB - A new procedure for the quantitation of C-21 ketosteroids using trifluoromethanesulfonic acid-catalyzed precolumn dansylation and coupled column liquid chromatographic separation, followed by postcolumn 1,1'-oxalyldiimidazole peroxyoxalate chemiluminescence detection is presented. In the simultaneous optimization of chromatographic resolution and chemiluminescence intensity, a coupled column chromatographic system and a stopped-flow system were used. An eluent containing 20 mM phosphate buffer at pH 6.7 accomplished an efficient separation of 3 alpha-hydroxy-5 beta-pregnan-20-one from a mixture containing 10 C-21 ketosteroids. Phosphate buffer also proved to be the most advantageous, among the six buffers tested, for sensitive detection. Experimental design and multivariate data analysis were used to characterize and optimize the postcolumn reaction chemistry in the chromatographic system. A valid full factorial design with excellent predictability showed that the flow rates for both 1,1' oxalyldiimidazole and hydrogen peroxide were the factors most strongly affecting the sensitivity of the system. The theoretical plate numbers were above 11,000 for all 10 dansylated ketosteroids. The 3 sigma detection limit estimated from 3 alpha-hydroxy-5 beta-pregnan-20-one calibration curve data was 1.6 pmol (n = 4, 125 microL injected) and spiked serum containing 0-74 pmol of this compound showed overall recoveries of 73 +/- 9% (n = 12). Quantitation of 3 alpha-hydroxy 5 beta-pregnan-20-one was finally carried out on 45 serum samples and the results compared to those from a radioimmunoassay (RIA) method. The data acquired with the procedure described in this work compare well with the results from RIA, which confirms the reliability of the new analytical procedure. PMID- 9852784 TI - RP-HPLC binding domains of proteins. AB - Procedures have been developed to identify the chromatographic binding domains of horse heart cytochrome c (Cyt c) and bovine growth hormone (bGH) during their interaction with reversed-phase sorbent materials. The procedure involves adsorption of the protein solute to the chromatographic sorbent, followed by proteolytic cleavage. Comparison of the proteolytic map obtained for Cyt c and bGH in free solution with the corresponding map obtained when these proteins are adsorbed to the chromatographic sorbent revealed significant differences in the digestion pattern. Following characterization of the peptides generated in both maps, the results indicated that specific regions on the surface of both Cyt c and bGH are inaccessible to tryptic cleavage when adsorbed to the hydrophobic surface of both a C-4 and a C-18 sorbent. Based on the assumption that the region of the protein surface that is in contact with the sorbent remains intact and bound to the sorbent during the digestion step, while the protein surface that is exposed to the solvent is accessible to proteolysis, the regions that were inaccessible to tryptic digestion were found to correspond to hydrophobic domains on the protein surface. These results also suggest that the three-dimensional structures of these proteins remain largely intact upon adsorption to the hydrophobic surface. PMID- 9852785 TI - Biosensor for direct determination of organophosphate nerve agents using recombinant Escherichia coli with surface-expressed organophosphorus hydrolase. 2. Fiber-optic microbial biosensor. AB - A fiber-optic microbial biosensor suitable for direct measurement of organophosphate nerve agents was developed. The unique features of this novel microbial biosensor were the recombinant Escherichia coli cells expressing the enzyme organophosphorus hydrolase on the cell surface and the optical detection of the products of enzyme-catalyzed organophosphate hydrolysis. The use of cells with the metabolic enzyme expressed on the cell surface as a biological sensing element provides advantages of no resistance to mass transport of the analyte and product across the cell membrane and low cost due to elimination of enzyme purification, over the conventional microbial biosensors based on cells expressing enzyme intracellularly and enzyme-based sensors, respectively. The use of an optical transducer allows the detection of different organophosphates in a mixture, presently not feasible with acetylcholinesterase-based biosensors. E. coli cells expressing organophosphorus hydrolase (OPH) on the cell surface were immobilized in low melting temperature agarose on a nylon membrane and attached to the common end of a bifurcated fiber-optic bundle. OPH-expressing E. coli cells catalyzed the hydrolysis of organophosphorus pesticides to form stoichiometric amounts of chromophoric products that absorb light at specific wavelengths. The backscattered radiation of the specific wavelength incident light was measured using a photomultiplier detector and correlated to the organophosphate concentration. The best sensitivity and response time were obtained using a sensor constructed with 1.5 mg of cells operating in pH 9, 50 mM HEPES buffer with 100 mM NaCl and 0.05 mM CoCl2 at 30 degrees C. At optimized conditions, the biosensor measured paraoxon, parathion, and coumaphos pesticides with high selectivity against triazine and carbamate pesticides in approximately 10 min. The lower detection limits were 3 microM for paraoxon and parathion and 5 microM for coumaphos. When stored in the buffer at 22 degrees C, the biosensor was stable for over a 1-month period and showed no decline in the response for over 75 repeated usages. The new fiber-optic microbial biosensor is an ideal tool for on-line monitoring of the detoxification process for organophosphate pesticides-contaminated wastewaters but may not be suitable for environmental monitoring. PMID- 9852786 TI - Immunosensor for 2,4-dichlorophenoxyacetic acid in aqueous/organic solvent soil extracts. AB - The development of a simple electrochemical immunoassay procedure for the field based quantification of the herbicide 2,4-D in methanolic soil extracts is presented. The sensor utilizes a competitive immunoassay format incorporating an immobilized antigen complex at the surface of a disposable screen-printed working electrode element. The extent of glucose oxidase-labeled antibody binding to the antigen--electrode is determined amperometrically and is related to sample analyte concentration. The performance of the sensor is assessed in buffer, 30% methanol, and methanolic soil extracts. The device is capable of quantifying 2,4 D in all three matrixes at the low ppm level with coefficient of variation values of 6.2-33.6%. The causes of the variation observed in the sensor response in different soil matrixes are examined and improvements proposed. The sensor, tested in parallel with a commercial 2,4-D immunoassay test kit, yields comparable quantitative data and detection limits while exhibiting greater assay simplicity. PMID- 9852787 TI - Electrodeposited iridium oxide pH electrode for measurement of extracellular myocardial acidosis during acute ischemia. AB - In the present paper, fabrication, characterization, and physiological applications of a solid-state pH electrode are described. The pH sensing layer was based on an anodic electrodeposited iridium oxide film (AEIROF). Sputtered platinum electrodes (1 mm diameter) fabricated on flexible Kapton films or platinum wires were used as planar or cylindrical supports. Each electrode site was coated with Nafion to attenuate the interference of anionic redox species and to protect the electrode surface during in vivo measurements. Performance of the AEIROF was evaluated, for the first time, as a pH electrode and proved to have a slightly super-Nernstian response with slope of -63.5 +/- 2.2 mV/pH unit for both wire and planar sputtered platinum electrodes. Linear pH responses were obtained in the pH range 2-10. The electrodes have a working lifetime of at least 1 month with accuracy of about 0.02 pH unit and fast response time. The electrodes showed very low sensitivities for different species, such as Na+, K+, Li+, NH4+, Ca2+, Mg2+, dissolved oxygen, lactate, ascorbate, and urate, which are important for physiological applications. The electrodes were applied in extracellular pH measurements during brief regional ischemia in a swine heart and no-flow ischemia in an isolated rabbit papillary muscle. A first report on extracellular pH, K+, and lactate simultaneous measurements during no-flow ischemia using the AEIROF pH electrode and the previously described K+ and lactate electrodes is presented as well. PMID- 9852788 TI - Fast-scan cyclic voltammetry of protein films on pyrolytic graphite edge electrodes: characteristics of electron exchange. AB - The rapid electron-exchange characteristics of metalloproteins adsorbed at a pyrolytic graphite "edge" electrode have been studied by analog dc cyclic voltammetry at scan rates up to 3000 V s-1. The voltammetry of four proteins, azurin (a "blue" copper protein) and three 7Fe ferredoxins, reveals oxidation and reduction peaks that display only modest increases in width and peak separation as the scan rate is raised. This is indicative of a substantially homogeneous population of noninteracting centers which undergo rapid electron exchange with the electrode. Both the Butler--Volmer and Marcus models have been tested. The electrochemical kinetics, as reflected by k0 (the rate at zero overpotential), are too fast to allow the determination of reorganization energies by this method. Nonetheless, the rapid and energetically coherent nature of the electron transfer enables the cyclic oxidation and reduction of protein redox centers to be examined on a time scale sufficiently short to recognize coupled processes occurring in the millisecond time domain, which are characteristic of the protein under investigation. Two of the ferredoxins display increasingly asymmetric voltammetry as the scan rate is increased, which is attributed to the coupling of electron transfer to conformational (or orientational) changes. For azurin, the use of higher electrolyte concentrations enables studies to be made at scan rates up to 3000 V s-1, from which a standard electron-transfer rate constant in the region of 5000 s-1 is obtained. At these high scan rates, azurin still shows very symmetrical voltammograms but with peak shapes displaying a more gradual decrease in current, at increasing overpotential, than is predicted using realistic values of the reorganization energy. The ability to measure even faster rate constants and access coupled reactions occurring in shorter time domains is likely to be limited by complex processes occurring on the graphite surface. PMID- 9852789 TI - Immunoelectrodes in protein detection: comparison between glassy carbon and a semimetallic Ni/P thin film as binding support. Biological applications. AB - Though immunoelectrodes can allow direct detection of very low protein amounts (about 0.1 pmol) in vitro and in vivo, they are not yet widely used because they need quality improvement. Based on a few works devoted to the basic electrochemical phenomenon occurring when antibodies are linked onto a solid support and during antigen/antibody complex formation, we have coated two different supports with antibodies: the classical glassy carbon fiber or an epoxy plate covered with an amorphous semimetallic (nickel/phosphorus) thin film obtained by means of an electrochemical deposit. The antibody/antigen complex formation induces direct and/or indirect ionic movements and a current flow through the conductive support toward a very low-noise and high-sensitivity preamplifier stage in an I/V configuration. The proposed electrochemical treatment (hydrophilization), applied to both carbon and Ni/P electrodes, improves antibody binding and reliability of the response to antigens. The Ni/P probes present several advantages when compared to carbon fiber: better conductivity, possibility of surface quality control, and semimetallic nature, making them unbreakable. Several applications were proposed: somatostatin-14 detection with both carbon fiber and Ni/P plate electrodes, and histamine detection in simple and complex fluid media. Dose-response curves and analysis of the results lead us to conclude that the obtained currents are directly related to the quantity of antigen. PMID- 9852790 TI - System for preparing microhybridization arrays on glass slides. AB - We describe the construction and operation of an arrayer system to produce patterns of DNA sequences for analytical uses such as microarrays of oligonucleotide on microchips. Detailed documentation on construction is provided, as well as added electronic circuitry and the software for the instrument, including programs to machine its own working surface as well as those to operate it as an arrayer. Its cost is modest, and with a single droplet tip it can deposit 96 spots per slide on 32 slides in about 200 min (readily upgraded to higher speeds). As currently operated, it can place 400 spots in 1 cm2, and this density, too, can be increased easily. We discuss design features and performance to demonstrate utility and flexibility. PMID- 9852792 TI - Yoga. The ultimate mind-body workout. PMID- 9852793 TI - When I tell my husband that the perpetual haze of pipe smoke in our house is hazardous, he says, "all those statistics refer to cigarette smoke, not pipe smoke." Is this true? PMID- 9852791 TI - Insomnia. Get a good night's sleep. PMID- 9852794 TI - Eye disorders. Spotting retinal detachment. PMID- 9852796 TI - Skin cancer and mortality. PMID- 9852795 TI - Prostate study favors surgery. PMID- 9852797 TI - Very low fat diets questioned. PMID- 9852798 TI - Implants and veneers. PMID- 9852799 TI - Dental implant outcomes in postmenopausal women undergoing hormone replacement. AB - To determine if hormone supplementation reduces the risk of failure for osseointegration of dental implants in postmenopausal women, the treatment outcomes of 116 women older than age 50, treated with 450 endosseous dental implants, were analyzed in this retrospective study. The findings indicated that hormone replacement therapy may not be linked with improved outcomes of endosseous dental implant treatment in postmenopausal women. Smoking, however, appears to significantly increase the implant failure rate in the group observed in this study. PMID- 9852800 TI - Oral sedation. AB - "I fear a trip to the dentist more than I fear death" is the response one person gave in a national survey recently cited in USA Today. While clearly representing an extreme, the results of many surveys suggest that fear of dentistry is still prevalent and is a measure of the failure of current therapeutic approaches to reduce pain and anxiety sufficiently to enable people, especially those with special needs, to visit the dentist. Patients who are fearful would likely seek oral health care more regularly if anesthesia and sedation were more readily available. Taking into consideration that the safety of anxiolytic drugs is highly dependent on the drug, dose, and route of administration used, oral premedication should be the sedative technique used by most dentists because it is efficacious, requires little monitoring when appropriate doses are used, and is unlikely to result in serious morbidity. PMID- 9852801 TI - Handling characteristics of resin composites in posterior teeth. AB - In the last 10 years, tremendous improvements in strength and shade selection for resin composites have been achieved. Also, a new generation of enamel-dentin bonding systems has been developed, and patient expectations of esthetic treatment have risen. Several techniques are available for restoring posterior teeth. When a caries lesion is limited, a direct esthetic restoration is indicated. Essential elements for obtaining good, long-term clinical results for direct esthetic restorations of posterior teeth are: (1) cavity preparation; (2) knowledge of the characteristics of the three dental substrates; (3) rubber dam use and matrix and wedge placement; (4) correct use of the enamel-dentin bonding system; (5) proper selection of the resin composite material; (6) use of the multilayering technique; (7) finishing and polishing procedures; and (8) maintenance of the restoration. PMID- 9852802 TI - The Spline implant interface: analysis and initial clinical experiences. AB - A new implant with a spline-shaped interface (Spline) was recently introduced. A common and preferred method of connecting two cylinders in many fields of industry, the Spline offers implant dentistry improved strength and precision compared to earlier and current implant connection designs. A comparative engineering analysis of the Spline and the major implant interfaces is offered, including the possible clinical implications. Initial surgical and restorative experiences with cement and screw-retained prostheses are presented in case reports. PMID- 9852803 TI - What do you mean, "My insurance did not pay"? PMID- 9852804 TI - Complications associated with diabetes mellitus after guided tissue regeneration: case report. AB - Dental practitioners need to be aware of the complications that can arise in the management and treatment of patients with diabetes mellitus. Patients with diabetes, and patients with a family history of diabetes, are at-risk dental patients. They are more likely to develop periodontal disease, and the periodontitis is more likely to be severe. Diabetes influences the progression and severity of periodontitis through changes in the small blood vessels, decreased collagen formation, and impairment of the host's defense mechanisms. Furthermore, complications associated with diabetes, such as impaired wound healing, can affect the patient's response to periodontal therapies like guided tissue regeneration (GTR). The case report in this article discusses the postsurgical complications that occurred during GTR treatment of a patient with non-insulin-dependent diabetes. The diabetic's susceptibility to periodontal disease and impaired wound healing can affect the progression of the disease and its treatment. Dental patients with diabetes require close supervision and frequent monitoring of their medical and dental health by the dental clinician. PMID- 9852805 TI - Underutilization of MRI. A suggested protocol. PMID- 9852806 TI - TMD treatment outcomes: a statistical assessment of the effects of psychological variables. AB - This study analyzes the degree to which pretreatment psychosocial factors (psychological dysfunctions and stress) effect outcome in 269 consecutive temporomandibular disorder (TMD) patients at the completion of treatment. Employing the TMJ Scale, a validated measure of TMD symptoms, it is found that pretreatment TMD pain and overall symptom levels (excluding internal derangement symptoms) are weakly but nevertheless, significantly related to pretreatment psychological dysfunction and stress. However, the latter appeared totally unrelated to four treatment outcome measures. Additionally, the data supports the hypothesis that both initial and post-treatment intrascapular symptoms (TMJ Scale, Joint Dysfunction sub-scale) are unrelated to psychosocial factors. Data from this study call into question the value of categorizing the TMD patients by means of psychosocial "profiling" and "dual-axis" classification methods proposed by some researchers. PMID- 9852807 TI - Exogenous estrogen may exacerbate thrombophilia, impair bone healing and contribute to development of chronic facial pain. AB - A 32 year old white female, in apparently good health, failed to respond to conservative wound care for alveolar osteitis after a routine mandibular first molar extraction. Curettage and biopsy of necrotic alveolar bone from the #30 socket escalated her pain such that hospitalization was necessary for pain management with intravenous morphine. Twelve months prior to admission she had been placed on exogenous estrogen (Premarin, 0.625 mg/day) after a partial oophorectomy. While hospitalized, she was found to have resistance to activated protein C (APCR). Premarin was discontinued. After discharge, weekly changes of an antibiotic impregnated dressing allowed for progressive regeneration of bone and epithelium with gradual reduction in her pain. She was found to be heterozygous for the mutant Factor V Leiden, a heritable factor for increased tendency to form thrombi, so-called thrombophilia. We speculate that the exogenous estrogen administration exacerbated the thrombophilia associated with the Factor V Leiden mutation by compounding the patient's resistance to activated protein C thereby contributing to her development of osteonecrosis and severe alveolar neuralgia. PMID- 9852808 TI - Risk factors and initial surgical failures of TMJ arthrotomy and arthroplasty: a four to nine year evaluation of 303 surgical procedures. AB - Studies exist which support the efficacy of TM joint arthrotomy, arthroplasty and arthroscopic surgery. Few, if any, studies have evaluated failures of arthroplasty and/or diskectomy and specific risk factors that might invite initial surgical failure. This paper is a retrospective review of 210 patients operated with arthrotomy/arthroplasty for painful and dysfunctional TM joint derangement. There were 303 surgical procedures evaluated over a follow-up period of 4-9 years. Patient ages ranged from 16-72 years. There was no age correlation seen with degree of joint derangement. All cases were operated by one surgeon. There were no cases of alloplastic materials in this group of patients. There were no cases of autograph such as auricular cartilage for dermal grafting or other disc substitution materials. Operations consisted of capsular arthroplasty in Wilkes' stage II, III, and IV. Diskectomy was performed in Wilkes' stages IV and V. Comparisons are made among staged groups and operation performed. Two hundred seventy-three of 303 operated joints met the criteria for surgical success for a technical success rate of 90.1%. Potential risk factors of missing molar teeth, preoperative joint collapse, and skeletal malocclusion were evaluated. The frequency of their presence in successful and non-successful surgical outcomes is noted. Patients with imaging confirmed osteoporosis were evaluated as group with potential systemic disease or a result of systemic disease that may influence long term surgical outcome. Predictable preoperative risk factors that may influence initial surgical outcome do appear to be significant in long term success. There were 30 cases of failure to evaluate. It is concluded that reconstructive arthroplasty is a stage specific operation with excellent results in Wilkes' stage II and good results in stage III derangement. Attempted arthroplasty failed significantly (50%) in a small number of attempts in stage IV cases. However, diskectomy was successful in stage IV and V cases. Osteoporosis may be the most significant risk factor and the presence of risk factors studied may jeopardize initial surgical outcomes. Preoperative staging of joint derangement is strongly suggested and evaluation of risk factors may necessitate selection of specific initial surgical procedures that minimize the influence of concomitant risk factors to long term success. PMID- 9852809 TI - Range of motion of the temporomandibular joint in rheumatoid arthritis: relationship to the severity of disease. AB - The purpose of this study was to determine the frequency and nature of temporomandibular joint (TMJ) involvement in rheumatoid arthritis (RA) patients with symptoms in this joint, and to investigate the relationship of symptoms to the C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and Steinbrocker stage. Clinical examination of the range of motion of the TMJ was performed in 218 RA patients. In addition, correlations between the maximal mouth opening and the severity of RA were studied. Restriction in opening the mouth (defined as < or = 30 mm movement in the central incisor region from the fully occluded to maximally open positions) was observed in 12.8% of the RA patients (28/218). The CRP, ESR and Steinbrocker stage were all correlated with maximal opening (p < 0.05). There was a positive correlation between the severity of RA and the range of motion of the jaw. PMID- 9852810 TI - Visual input effect on EMG activity of sternocleidomastoid and masseter muscles in healthy subjects and in patients with myogenic cranio-cervical-mandibular dysfunction. AB - This study was conducted in order to determine the input visual effect on electromyographic (EMG) activity of the sternocleidomastoid and masseter muscles in the supine and lateral decubitus positions. The study was performed on 22 patients with myogenic cranio-cervical-mandibular dysfunction (CMD) and 18 healthy subjects. EMG activity at rest and during swallowing of saliva and maximal voluntary clenching was recorded in the supine and lateral decubitus positions in the following conditions: 1. with eyes open; and 2. with eyes closed after 5 minutes in a dark room. A significant decrease of EMG activity at rest with closed eyes in both groups was observed in the sternocleidomastoid (lateral decubitus position) and in the masseter muscle (supine position). During swallowing of saliva a significant decrease of EMG activity with closed eyes was observed only in the sternocleidomastoid muscle (lateral decubitus position) in healthy subjects. During maximal voluntary clenching any significant differences were observed upon variation in the visual input. The significant change in EMG activity, mainly observed at rest, suggests that the visual input effect is weak. The absence of a significant change in EMG activity during maximal voluntary clenching upon variation in the visual input could be clinically relevant in patients with myogenic CMD who habitually brux. PMID- 9852811 TI - Health care utilization by patients with temporomandibular joint disorders. AB - The claims data base of a large New England managed care organization was used to compare the health care utilization patterns of patients with TMJ disorders to non-TMJ subjects. Inpatient, outpatient and psychiatric claims data were examined over a wide range of diagnostic categories. Age and sex adjusted results showed that, overall, patients with TMJ disorders were greater utilizers of health care services and had higher associated costs than non-TMJ subjects. For some of the major diagnostic categories, such as nervous, respiratory, circulatory, and digestive, the inpatient and outpatient claims differences in utilization and costs were as large as 3 to 1. For only one diagnostic category, pregnancy and childbirth, were utilization and costs greater for non-TMJ subjects than TMJ patients. The psychiatric claims for TMJ patients exhibited differences that were at least twice as large as those for the non-TMJ subjects. PMID- 9852812 TI - Cervical signs and symptoms in patients with Meniere's disease: a controlled study. AB - This study compares the frequency of signs and symptoms from the cervical spine in 24 patients diagnosed with Meniere's disease and 24 control subjects from a population sample. From a previous controlled comparative study concerning signs and symptoms of craniomandibular disorders, 24 patients diagnosed with Meniere's disease (10 males and 14 females) and their 24 matched control subjects participated in this investigation on the state of the cervical spine. Symptoms of cervical spine disorders, such as head and neck/shoulder pain, were all significantly more frequent in the patient group than in the control group. Most of the patients (75%) reported a strong association between head neck movements in the atlanto-occipital and atlanto-axial joints and triggered attacks of vertigo. Also, 29% of the patients could influence their tinnitus by mandibular movements. Signs of cervical spine disorders, such as limitations in side-bending and rotation movements, were significantly more frequent in the patient group than in the control group. Tenderness to palpation of the transverse processes of the atlas and the axis, the upper and middle trapezius, and the levator scapulae muscle were also significantly more frequent in the patient group. The study shows a much higher prevalence of signs and symptoms of cervical spine disorders in patients diagnosed with Meniere's disease compared with control subjects from the general population. PMID- 9852813 TI - A skull base epidermoid cyst causing the symptoms of a craniomandibular disorder. AB - This article describes a case report of a 52 year old patient with a ten year history of orofacial pain who was misdiagnosed with a craniomandibular disorder (CMD) and trigeminal neuralgia. After a epidermoid cyst in the skull base had been diagnosed and removed the complaints diminished and finally disappeared. PMID- 9852814 TI - Sports-related dental injuries and sports dentistry. PMID- 9852815 TI - Exceeding expectations: may the workforce be with you the significant other ... the dental assistant. PMID- 9852816 TI - Professional and predictable home bleaching. PMID- 9852817 TI - Basic computing for dental practitioners: 1. The principles of computers and computing. AB - The purpose of this series of six articles is to introduce the interested general dental practitioner to computers and computing, to remove much of the mystery surrounding computers and to explain the technology in straightforward terms. The first two articles will concentrate on the basic principles; later papers will discuss the most commonly used software, practice management systems and multimedia. PMID- 9852818 TI - An update on antimicrobial chemotherapy: 1. The mechanisms of action of antibiotics. AB - This three-part series will provide an update on the modes of action of currently available antibiotics, review the mechanisms of antibiotic resistance and discuss the impact of antibiotic resistance on prescribing for oral infections. PMID- 9852819 TI - An overview of dental care for the young patient: 3. Treatment measures. AB - In this final part of the series the reasons for providing high-quality restorations for the young patient are considered. For such treatment to be successful it must be underpinned by effective management of the child's behaviour. Appropriate techniques and materials for a range of commonly encountered situations are discussed, with particular emphasis on the restoration of carious teeth. PMID- 9852820 TI - Malignant melanoma of the head and neck. AB - Malignant melanoma is rapidly increasing in incidence. The primary lesion is frequently seen in the head and neck and general dental practitioners thus have the opportunity to identify suspicious lesions at an early stage. This article gives an overview of the pathology and clinical features of melanoma. PMID- 9852821 TI - Papillon-Lefevre syndrome: a case report. AB - This case report discusses a very rare condition; Papillon-Lefevre Syndrome. This is an autosomal recessive condition in which the main features are hyperkeratosis of the palms and the soles and severe periodontal destruction. PMID- 9852822 TI - Young adult patients with established dental erosion--what should be done? AB - This clinical paper discusses the problem of managing established erosion in the young patient. For a significant number of these patients it is not possible to establish conclusively the erosive source. Intermediate composite restorations, in addition to the use of normal preventive measures, can be used to protect eroded teeth from further damage. The main goal of such restorations is to protect the viability of the worst-affected teeth until such time as the aetiological factors can be identified and controlled. They can also improve overall aesthetics and create interocclusal space. PMID- 9852823 TI - An update on antimicrobial chemotherapy: 3. Antimicrobial resistance and the oral cavity. AB - As mentioned in the preceding articles, the widespread appearance of antimicrobial resistance has become a major threat to the effective treatment of infectious disease. However, does this have any relevance to the dental profession? This article relates current knowledge to everyday dental practice. PMID- 9852824 TI - Treatment of loss of tooth substance using dentine-bonded crowns: report of a case. AB - This article discusses the treatment of a bulimic patient with severe tooth substance loss. The patient was provided with dentine-bonded crowns in view of the need for only minimal tooth preparation, their potential for good aesthetics and good patient acceptance. The crowns have performed satisfactorily in clinical service for 4 years. PMID- 9852825 TI - Basic computing for dental practitioners: 3. Word processors. AB - The three computer applications most widely used on personal computers are word processors, spreadsheets and databases. A word processor is essentially a program designed to create, print and store documents and letters. Microsoft Word and Lotus WordPro are examples. The term spreadsheet is used to describe a type of program that is designed to undertake financial, mathematical and even statistical calculations on rows and columns of numeric data. A spreadsheet program is useful for maintaining practice accounts, calculating simple statistics and trying out 'what if' scenarios on numerical data. A database is analogous to a card index system, and is used to store information in a format that facilitates rapid retrieval of data using a multitude of search methods. Many databases are capable of processing this information to generate data summaries, reports and charts based on the search criteria. In this article the basic features of word processing packages are described. Spreadsheets and databases will be discussed in a later paper. PMID- 9852826 TI - A review of dentifrices. AB - Dentifrices have a role to play in the prevention of certain oral diseases, such as dental caries and gingivitis. The use of fluoridated dentifrices has had a dramatic effect on the caries rate in the western world over the last 30 years, and antibacterial, desensitizing and anticalculus dentifrice formulations have recently been developed. This paper provides a brief overview of the dentifrices currently available. PMID- 9852827 TI - Diver's mouth syndrome: a report of two cases and construction of custom-made regulator mouthpieces. AB - Scuba diving is becoming more and more popular and dentists are increasingly likely to encounter patients with diving-related problems. Protruded mandibular positions and the biting forces exercised on the anterior occlusion during diving can cause pain and dysfunction, particularly where a predisposing factor such as bruxism exists. In this article, two cases are reported of patients with diving exacerbated pain dysfunction syndrome, which improved following the construction of custom-made mouthpieces which distributed the forces more evenly. Laboratory procedures are described for construction of a custom-made scuba regulator mouthpiece that allows more comfortable diving. PMID- 9852828 TI - The MFDS/MFD examination. AB - The MFDS is a new examination designed and administered by the Royal Surgical Colleges of England, Scotland and Ireland. It can be taken after GPT and will be a pre-requisite to the commencement of specialist training. PMID- 9852829 TI - Economic evaluation in dentistry: an ethical imperative? AB - In this paper a case is made for the use of principles and techniques of economic evaluation to aid the planning of oral health services. The reader is taken through a case study, which highlights not only the importance of economic principles but also the challenges involved in carrying out economic evaluations in dentistry. These challenges need to be faced if oral health services are to be planned in a way that is of maximum benefit to the community. PMID- 9852830 TI - A practical approach to presurgical orthodontic preparation. PMID- 9852831 TI - JCO interviews. Samir E. Bishara, BDS, DOrtho, DDS, MS on growth and orthodontic treatment. PMID- 9852832 TI - A safer debonding/debanding technique. PMID- 9852833 TI - Modified arrowhead clasps for removable biteplanes. PMID- 9852834 TI - The Mandibular Protraction Appliance No. 3. PMID- 9852835 TI - Use of nickel titanium coil springs for partially impacted second molars. PMID- 9852836 TI - Frictional resistance of the Damon SL bracket. PMID- 9852837 TI - Simplified activation of closing loops. PMID- 9852838 TI - The universal bite jumper. PMID- 9852839 TI - A method for light-cured indirect bonding. PMID- 9852840 TI - Orthodontic management of a dilacerated maxillary central incisor with an unusual sequela. PMID- 9852841 TI - Bond strengths of two resin-modified glass polyalkenoate cements under different surface conditions. PMID- 9852842 TI - Orthodontic marking pencils: a potential source of cross-contamination. PMID- 9852843 TI - Bonded acrylic lingual biteplanes. PMID- 9852844 TI - The lingual pearl. PMID- 9852845 TI - A quick and easy diagnostic setup technique. PMID- 9852846 TI - CDA hones its marketing message. AB - This article discusses the many tacks CDA has taken with its marketing campaigns over the years. The keys to success have been reasonable goals and sufficient funds to make an impact. The current campaign, while underfunded due to budget constraints, carries on the successful theme of positioning CDA and its member dentists as the trusted sources of dental information for consumers, legislators, and health care decision-makers. An accompanying article discusses the proposed ADA national marketing campaign. PMID- 9852847 TI - Making people want dentistry. AB - With millions of dollars being spent by advertisers seeking a share of consumers' disposable income, dentistry is up against fierce competition for patients. Dentistry will have to sell itself to keep up, and one way to do that is to focus on the area in which consumers are increasing their pending-esthetics. Once patients are drawn into a practice by appeals to their desire to look good, they can then be assessed for necessary work to improve their overall dental health. PMID- 9852848 TI - The dynamics of a village: marketing strategies for a new millennium. AB - Successful marketing of a dental practice involves more than a yellow pages ad. picking a dentist is an emotionally based decision. Dentists who take steps to become involved in their communities and interact with the people within them will reap the personal and professional rewards necessary for a successful practice. PMID- 9852849 TI - Marketing on a tight budget. AB - Marketing is necessary to build a dental practice. A consistent, repeating, and ongoing set of marketing strategies is necessary to put a practice on the map and get people excited about becoming patients. Often, however, a dentist doesn't realize the necessity of marketing until such efforts and least affordable. This paper discusses a variety of low-cost ways of marketing a dental practice. PMID- 9852850 TI - Using public relations for a dental practice. AB - Public relations is a way of marketing a dental practice through the media and directly to potential patients without purchasing advertising time or space. Well written press releases and follow-up phone calls targeted to the specific audience of various media outlets can result in stories that are worth more than purchased advertising. One key to successfully communicating to others through public relations is straightforward writing. The following article not only covers the variety of opportunities available to a dental practice through PR, it also serves as an excellent example of the style of writing that should be used for PR communications. PMID- 9852851 TI - Snoring and obstructive sleep apnea from a dental perspective. AB - Proper diagnosis and treatment of sleep-related disorders are best handled via a team approach. This team may include a general dentist treating in conjunction with other sleep specialists. However, to provide care, dentists must have a basic understanding of sleep disorders. This paper provides a dental overview of sleep-related breathing disorders, including key definitions, an outline of a diagnostic protocol, a discussion of the factors involved in decision-making, and a summary of the wide variety of treatment modalities. PMID- 9852852 TI - Diagnostic sleep testing in the evaluation of snoring and sleep apnea. AB - The dilemma frequently encountered by the dentist wishing to offer an appliance for treatment of snoring or sleep apnea is whether there is a need for extensive testing of patients who present with a complaint of snoring. This article describes diagnostic sleep testing in the evaluation of patients with snoring and sleep apnea. PMID- 9852853 TI - Medical and nondental treatments of snoring and sleep apnea syndrome. AB - The prevalence, pathophysiology, and clinical and polysomnographic evaluation of obstructive sleep apnea are reviewed. The history of the development of nasal continuous positive airway pressure, diagnostic titration of the treatment, abolition of nocturnal apneas, and consolidation of sleep architecture by nasal continuous positive airway pressure and long-term patient compliance with the treatment are discussed. The effects of weight gain and weight loss on the severity of obstructive sleep apnea, and cardiovascular and behavioral complications of obstructive sleep apnea are reviewed. Behavioral treatments are discussed. PMID- 9852854 TI - Surgical options for obstructive sleep apnea. AB - The pathology associated with obstructive sleep apnea is cumulative and progressive. When patients fail to improve with continuous nasal airway pressure or other, less-invasive treatments, surgery should be considered. The initial approach to the surgical patient is identification of all areas of potential obstruction. There are often several sites of obstruction, which can occur anywhere in the upper respiratory tract. One or more procedures may be needed to address these areas. The objective of surgery is to relieve these obstructing sites without interfering with the normal functionality of the upper airway. PMID- 9852855 TI - Dentistry's role in the recognition and treatment of sleep-breathing disorders: the need for cooperation with the medical community. AB - While oral appliance therapy for the treatment of sleep-disordered breathing can be an exciting and rewarding adjunct to the practice of dentistry, it is essential that dentists realize that snoring and obstructive sleep apnea are medical and not dental problems. Sleep-disordered breathing and its sequelae are diseases that should remain in the purview of the medical community. While the dentist can identify patients with sleep-breathing disorders and participate in their treatment, it is essential to emphasize that sleep-breathing disorders are potentially life-threatening diseases whose diagnosis and treatment are the domain of the medical profession. Accepting dentistry's position as part of a treatment team, ongoing review of scientific literature, cooperation with medical colleagues, and attendance at educational meetings dedicated to the study of sleep-related disorders are essential to proper and ethical dental participation in the treatment of sleep-disordered breathing. PMID- 9852856 TI - Should the dentist independently assess and treat sleep-disordered breathing? AB - Sleep-disordered breathing is a chronic problem of the inappropriate mechanical collapse of the upper airway. Symptoms range from mild occasional snoring to severe obstructive sleep apnea. The standard of care for the diagnosis and treatment of sleep-disordered breathing by sleep medicine has been the use of the polysomnogram and continuous positive airway pressure. This approach is burdensome, costly, and ineffective due to lack of compliance with or rejection of treatment. Oral appliances are highly effective in managing the mild snorer to the moderate sleep apneic and are approaching the efficacy of continuous positive airway pressure with the severe apneic. The dentist can and should manage these patients. However, the dental practitioner must acquire sufficient training and knowledge to appropriately treat these patients. PMID- 9852857 TI - Comparisons of oral devices for snoring. AB - A study was performed on 19 types of oral appliances for snoring/obstructive sleep apnea. The appliances were categorized into two groups, mandibular advancers and tongue advancers. A set of limited criteria was developed by which an appliance could be evaluated, and the criteria were weighted. Evaluations were then performed on the appliances, and they were rated according to satisfaction of criteria. The rating table should help a dentist in selecting an appliance that will be accepted by the patient and effective in treating snoring/obstructive sleep apnea. PMID- 9852858 TI - Telehealth technology: a new medium for in-service education. PMID- 9852859 TI - CDA information bulletin: dental unit waterlines. Canadian Dental Association, Board of Governors. PMID- 9852860 TI - The use of tongue cleaners in the treatment of halitosis. PMID- 9852862 TI - Identity crisis. PMID- 9852861 TI - Case #7: Reticular lichen planus. PMID- 9852863 TI - The trauma of cancer. PMID- 9852864 TI - Working for her. PMID- 9852865 TI - Former Fen-Phen users may need prophylactic antibiotic before treatment. PMID- 9852866 TI - Case #5. Hyperplastic candidiasis. PMID- 9852868 TI - Nutritionist discovers outlet for combining her careers. PMID- 9852867 TI - Beyond the veggies. PMID- 9852869 TI - Managing managed care. PMID- 9852870 TI - A patient's complicated medical history calls for a great deal of analytical thinking. PMID- 9852871 TI - Florida transplant shares her homework from hygiene board exams with peers. PMID- 9852872 TI - Bilateral internal thoracic artery grafting may improve outcome of coronary artery surgery. Risk-adjusted survival. AB - BACKGROUND: Excellent clinical results of the patients with single left internal thoracic artery (ITA) grafting compared with saphenous vein grafting have prompted surgeons to use both ITAs. However, long-term benefits of the bilateral ITA grafting have not been proven. METHODS AND RESULTS: We reviewed our experience of 2826 patients (age 62 +/- 9 years [mean +/- 1 SD], 2350 men, mean follow-up 52 months) who underwent surgery with ITAs, supplemented by saphenous vein grafts when required, between 1985 and 1995. Single ITA grafting (n = 1557) was compared with double (n = 1269), by means of the Cox proportional hazards model. Significant predictors of all-cause mortality were as follows: (1) peripheral vascular disease, rate ratio (RR) = 2.4 (1.7 to 3.4 [95% CI]); (2) prior myocardial infarction, RR = 2.1 (1.5 to 3.1); (3) severe left ventricular dysfunction, RR = 3.9 (2.6 to 5.9) and moderate left ventricular dysfunction, RR = 2.0 (1.5 to 2.6); (4) age > or = 70 years, RR = 3.4 (2.4 to 4.8), and age 60 to 69 years, RR = 1.7 (1.3 to 2.4); (5) diabetes mellitus, RR = 1.7 (1.3 to 2.4); (6) carotid disease, RR = 1.7 (1.2 to 2.4); and (7) single ITA (versus bilateral ITA), RR = 1.4 (1.1 to 1.8). Number of vessels diseased, surgical status (i.e., urgent versus elective), hypertension, and sex were not significant predictors. Unadjusted actuarial survival rates at 10 years were 86 +/- 3% (mean +/- 95% CI) for bilateral ITA group and 71 +/- 5% for single ITA. Single ITA was also a predictor of all-cause mortality, late myocardial infarction, or late reoperation (RR = 1.3 [1.1 to 1.6]). CONCLUSIONS: Bilateral ITA grafting may improve long term survival and freedom from late myocardial infarction or reoperation after coronary artery surgery. Mathematical modeling may assist in developing a strategy for use of bilateral ITA grafts. PMID- 9852873 TI - Multiple arterial grafts. Radial versus right internal thoracic arteries. AB - BACKGROUND: Left internal thoracic artery (LITA) grafts to the left anterior descending coronary artery (LAD) during coronary bypass surgery (CABG) have greater patency rates than saphenous vein grafts and reduce long-term cardiac morbidity and mortality rates. The benefits of multiple versus single arterial grafts and the role of different arterial conduits with respect to short- and medium-term outcome remains controversial. The purpose of this study was to compare the perioperative and intermediate-term results of: (1) patients receiving 2 arterial grafts versus 1 arterial graft and (2) patients receiving a right internal thoracic artery (RITA) versus a radial artery (RA) as the second arterial graft. METHODS AND RESULTS: Retrospective analysis of prospectively gathered data on consecutive patients undergoing isolated CABG at our institution between 1989 and 1996 was conducted. The first section of the study compared outcomes for 1 arterial graft (LITA to LAD, n = 2333) versus 2 arterial grafts (LITA + RA or LITA + RITA, n = 378). The second section of the study compared outcomes for the RITA (n = 132) versus the RA (n = 171) as second arterial grafts since 1992, when the radial series was initiated. Part I: By multivariable stepwise logistic regression, the use of 1 arterial graft was associated with an increased incidence of perioperative cardiac morbidity and mortality (odds ratio 2.2, 95% confidence interval 1.4 to 3.3), with the use of our current patient selection criteria. Double-arterial graft patients had a nonsignificant trend toward increased intermediate-term actuarial survival (P = 0.12) and cardiac event-free survival (P = 0.09). Part II: Comparison of preoperative demographics revealed a higher incidence of diabetes (27% vs 11%, P < 0.001), peripheral vascular disease (16% vs 8%, P = 0.03), and elderly age (13% vs 2%, P = 0.001) in patients receiving an RA versus those receiving a RITA as the second arterial graft. Perioperative outcome analysis revealed a decreased intensive care unit stay in the RA versus RITA group (median 30.4 vs 36.2 hours, respectively, P = 0.005) but no significant difference in hospital length of stay. There was no significant difference in perioperative mortality or cardiac morbidity rates. RITA patients had a higher incidence of sternal wound infection (5.3% vs 0.6%, P = 0.01), however, and tended to have increased blood product transfusion rates (51% vs 40%, P = 0.06). CONCLUSIONS: The use of 2 arterial grafts is safe, with a reduction in perioperative cardiac morbidity or mortality rates compared with 1 arterial graft after adjustment for other risk variables. When comparing RITA to RA as second arterial grafts, patients receiving an RA have a lower incidence of sternal wound infection and decreased transfusion requirements, with no difference in perioperative or intermediate-term cardiac morbidity or mortality rates. PMID- 9852874 TI - New approaches to prevention and treatment of radial artery graft vasospasm. AB - BACKGROUND: There has been renewed interest in radial artery (RA) conduits for coronary artery bypass because of the relative resistance of arterial grafts to atherosclerosis compared with autogenous vein grafts. Although improved drug therapy for arterial spasm is now available, vasospasm still occurs in at least 5% to 10% of RA grafts. We systematically evaluated the effectiveness of calcium channel blockers and organic nitrates for inhibition or reversal of RA contraction in vitro. Additionally, we investigated the efficacy of novel gene therapy with endothelial nitric oxide synthase (eNOS) to inhibit RA contractions. METHODS AND RESULTS: Segments of RA from 28 patients undergoing coronary artery bypass grafting were mounted in organ chambers. In control experiments, KCl (5 to 50 mmol/L) produced dose-dependent increases in tension (maximum tension, 14.3 +/ 3.0 g, n = 7). Addition of diltiazem or verapamil had no significant effect on KCl contraction (128 +/- 36% and 88 +/- 24% control, respectively); however, nifedipine markedly inhibited KCl contraction (27 +/- 4% control, P = 0.005). Norepinephrine (NE, 10(-9) to 10(-4) M) produced dose-dependent increases in tension (maximum tension, 15.7 +/- 2.7 g in control rings, n = 8). Diltiazem and verapamil pretreatment had no significant effect on NE contraction (103 +/- 14% and 90 +/- 14% control, respectively); nifedipine significantly inhibited NE contraction (70 +/- 11% control, P = 0.02). Isosorbide dinitrate and nitroglycerin markedly inhibited KCl contractions (47 +/- 9% and 30 +/- 8% of controls, n = 6) and NE contractions (42 +/- 10% and 31 +/- 9% of controls, n = 6). Nifedipine, isosorbide, and nitroglycerin were further evaluated for the ability to reverse an established contraction (KCl 40 mmol/L); nitroglycerin was most effective in reversing RA contraction. In separate experiments, RA underwent adenoviral-mediated gene transfer with vehicle, Escherichia coli beta galactosidase, or eNOS (eNOS, 10(10) PFU/mL x 1 hour). Transgene expression was confirmed by beta-galactosidase activity and eNOS immunohistochemistry after 40 hours of ex vivo incubation. Immunohistochemistry demonstrated recombinant NOS in adenovirus encoding bovine eNOS (Ad.CMVeNOS) RA only. Ad.CMVeNOS arteries contracted only 46.6 +/- 13.7% of controls to KCl (n = 5) and 48.2 +/- 11.4% of controls to prostaglandin F2 alpha a (10(-9) to 10(-6) M, n = 5). CONCLUSIONS: Diltiazem, which is used empirically to prevent RA vasospasm, had little effect on human RA contractions (receptor-independent and receptor-dependent). Organic nitrates inhibited and reversed RA contractions. Adenoviral transfer of NOS suggests that future clinical application of gene therapy may play an important role in prevention of RA vasospasm. PMID- 9852875 TI - Improving cost and outcome of coronary surgery. AB - BACKGROUND: There has been increasing interest in improving the outcome of coronary surgery while also seeking to minimize cost. It was the purpose of the present study to determine changes in the outcome and cost of CABG between 1988 and 1996. METHODS AND RESULTS: The outcome and costs for 12,266 patients undergoing CABG were evaluated. Clinical data were gathered from the Emory Cardiovascular Database, and financial data were obtained from the UB92 formulation of the hospital bill. Charges were reduced to cost through the use of departmental cost-to-charge ratios. Costs were inflated to 1996 costs by using the medical care inflation rate. The patients became sicker, especially with increased incidences of hypertension, diabetes, and prior myocardial infarctions and a decrease in ejection fraction over the study period. Mortality rates tended to decrease from 4.7% to 2.7% (P = 0.07). After accounting for increasing indexes of severity of disease over the period, there was a significant decrease in death (OR, 0.90/y; P = 0.0001). Q-wave myocardial infarction rate fell from 4.1% to 1.3% (P < 0.0001). Mean hospital cost decreased from $22,689 to $15,987. Length of stay after surgery decreased from 9.2 to 5.9 days. After accounting for other variables, cost decreased by $1118 per year, and annual length of stay decreased by 0.55 day. CONCLUSIONS: The outcome of CABG continues to improve with declines in mortality rate and Q-wave myocardial infarction. This was accomplished while decreasing costs and length of stay. Whether these favorable trends will continue remains to be seen. PMID- 9852876 TI - Impact of managed waiting for coronary artery bypass graft surgery on patients' perceived quality of life. AB - BACKGROUND: Current demand for CABG surgery remains high, often exceeds available resources, and has led to the development of managed waiting lists. This study was designed to determine how being placed on a managed waiting list for > 6 weeks for CABG surgery affected patients' perceived quality of life in a Canadian center. METHODS AND RESULTS: Telephone interviews were carried out in the setting of a large urban hospital in northern Alberta. All participants were identified from 3 waiting lists of adult patients waiting for open heart surgery. A master list of patient statements was compiled to formulate the 47-item Waiting List Impact Questionnaire (WLIQ). A total of 102 patients completed the WLIQ by telephone interview. Patients (87.5%) indicated that their quality of life had deteriorated since being placed on the waiting list. None of the patients perceived an improvement in their quality of life. Frequency data for the WLIQ provided a broad, multidimensional perspective of the experience of waiting for CABG surgery and its impact on perceived quality of life. Negative impact was found in each of 5 main themes: employment and income, physical stress, social support, frustration, and quality of life. CONCLUSIONS: This study indicates that patients perceived a negative impact on their quality of life after being placed on a managed waiting list for CABG surgery. In the allocation of healthcare resources, attention should be paid to the impact of waiting on patients' physical well-being as well as on quality of life. PMID- 9852877 TI - Hospital readmission after cardiac surgery. Does "fast track" cardiac surgery result in cost saving or cost shifting? AB - BACKGROUND: Intense medical and economic pressures have created "fast track" cardiac surgery in which clinical services are streamlined and early discharge is encouraged. Does this strategy promote significant cost saving or merely cost shifting? In a global system of reimbursement, the economic benefit of decreasing patient length of stay may be offset by high rates of patient readmission. This study was undertaken to determine the 30-day readmission rate after cardiac surgery and to analyze trends of readmission diagnoses. METHODS AND RESULTS: From October 1, 1996 to July 31, 1997, 460 consecutive cardiac surgical operations were performed at 1 institution. There were 25 deaths and 8 patients who remained as inpatients at the 30-day postoperative deadline for readmission. Two patients had 2 operations. Therefore, 527 operations were performed on 525 patients. There were 110 readmissions after 527 operations for a readmission rate of 20.9%. A significant number of readmissions (49%) were to outside hospitals. Readmission diagnoses were: atrial fibrillation (23%); angina, congestive heart failure, or ventricular tachycardia (20%); leg wound (15%); sternal wound (5%); pneumonia (5%); gastrointestinal complaints (5%); neurologic event (2%); and miscellaneous (25%). Patients discharged > or = 7 days postoperatively were twice as likely to be readmitted as those discharged on postoperative days 4, 5, or 6. CONCLUSIONS: Readmission after cardiac surgery is common and frequently (49%) to outside institutions. Patients discharged > or = 7 days postoperatively represent the patients at greatest risk of readmission and, therefore, warrant closer scrutiny before discharge. PMID- 9852878 TI - Does age limit the effectiveness of clinical pathways after coronary artery bypass graft surgery? AB - BACKGROUND: Clinical pathways have been shown to be effective in reducing the length of hospital stay after isolated CABG. Few studies, however, have focused specifically on the outcomes of the pathways in regard to the elderly population. METHODS AND RESULTS: We reviewed our experience with 445 consecutive patients (299 < 70 years old [mean age, 58.2 +/- 0.5 years] and 146 > or = 70 years old [mean age, 75.6 +/- 0.3 years]) who underwent isolated CABG with the expectation of progressing through the same 5-day postoperative pathway. Preoperatively, the elderly had a smaller body surface area (1.87 +/- 0.02 versus 2.00 +/- 0.01; P < 0.001) and a higher incidence of female gender (45.9% versus 26.8%; P = 0.001), cerebrovascular disease (13.7% versus 7.0%; P = 0.022), congestive heart failure (22.6% versus 13.4%; P = 0.013), and 3-vessel coronary artery disease (76.7% versus 65.9%; P = 0.024). Postoperatively, the elderly had a higher incidence of red blood cell transfusion (28.8% versus 9.0%; P = 0.001), atrial fibrillation (37.6% versus 11.7%; P = 0.001), and overall rate of complications (46.6% versus 23.4%; P = 0.001). Mortality rate and length of stay were 5.5% and 7.9 +/- 0.4 days for the elderly versus 1.0% and 6.4 +/- 0.4 days for those < 70 years old (P = 0.004 and P = 0.008), respectively. Of those > or = 70 years old, 34% were discharged in < or = 5 days, 64% in < or = 7 days, and 82% in < or = 10 days versus 64%, 85%, and 93%, respectively, for younger patients (P = 0.001 for all). Multivariate analysis of preoperative variables identified age (P < 0.001), female gender (P < 0.001), hypertension (P = 0.017), chronic obstructive pulmonary disease (P = 0.002), preoperative intra-aortic balloon pumping (P = 0.002), and body surface area (P = 0.003) as significantly related to length of stay. However, when the postoperative variables found to be different by univariate analysis are added to the model, age is only marginally significant (P = 0.079), and red blood cell transfusion and atrial fibrillation are the strongest predictors of increased length of stay, along with intra-aortic balloon pumping and pneumonia (P < 0.001 for all). CONCLUSIONS: These data suggest that extraordinary modifications of clinical pathways are not needed for success with elderly patients. The increased length of stay is largely attributable to the increased incidence of atrial fibrillation. PMID- 9852879 TI - Effect of payer status on outcomes of coronary artery bypass surgery in blacks. AB - BACKGROUND: Black patients with coronary artery disease have a higher mortality rate than white Americans. They also have a higher prevalence of hypertension, diabetes mellitus, and renal disease, which may have an effect on mortality rates. The deleterious effect of these comorbidities may be exacerbated by impaired access to secondary prevention strategies and longitudinal care. Therefore, the presence or absence of comprehensive care as indicated by payer status may then affect survival on surgically treated patients. In this study we examined the role of cardiovascular risk factors and insurance carrier status on early outcomes of coronary artery bypass grafting (CABG) surgery in blacks versus white Americans. METHODS AND RESULTS: From January 1990 to December 1996, 2776 patients (2003 men, 773 women; mean age 63 +/- 10 years), underwent isolated CABG in a multispecialty practice serving a major metropolitan population. There were 494 (17.8%) black patients and 2282 (82.2%) white patients. The proportion of black patients in each payer category was 17.8% commercial, 14.1% managed care, 52.9% Medicaid, and 19.5% Medicare. The effect of preoperative risk factors, including status of operation (elective, urgent, or emergent), sex, race, redo CABG, presence of renal disease, diabetes mellitus, congestive heart failure, myocardial infarction, the completeness of revascularization, age, and left ventricular ejection fraction were analyzed with the chi 2 test for categorical variables and the Student t test for age and ejection fraction. A multiple logistic regression analysis was performed to assess the effect of all variables on mortality rates simultaneously. Black patients had a higher incidence of diabetes mellitus, hypertension, and renal disease than white patients (P < 0.001). Overall, 30-day mortality rate was 2.5% (58 of 2282) in white patients versus 5.5% (25 of 494) for black patients (P < 0.003). Multivariate analysis showed that only emergency surgery status (OR 3.59, P < 0.01), redo CABG (OR 3.78, P < 0.001), hypertension (OR 2.32, P < 0.03), history of congestive heart failure (OR 2.1, P < 0.004), older age (OR 1.07, P < 0.001), and low ejection fraction (OR 0.98, P < 0.003) correlated with mortality rates. Race and payer status were not significant predictors of death. CONCLUSIONS: These data on CABG surgery in black patients suggest that early death is due to associated risk factors and not due to race or insurance payer status. PMID- 9852881 TI - Presence of angiographic coronary collaterals predicts myocardial recovery after coronary bypass surgery in patients with severe left ventricular dysfunction. AB - BACKGROUND: Patients with coronary artery disease and left ventricular dysfunction (LVD) may have areas of hibernating myocardium that improve functionally after revascularization. Coronary collateral circulation may sustain ischemic, dysfunctional myocardium and favor myocardial recovery after revascularization. We evaluated the effect of angiographic coronary collaterals on myocardial functional recovery after coronary bypass graft (CABG) surgery in a group of patients with severe LVD. METHODS AND RESULTS: Forty-one patients with multivessel coronary artery disease and advanced LVD (left ventricular ejection fraction [LVEF] 25 +/- 5%) undergoing CABG were identified from a prospective database. Preoperative coronary angiograms were evaluated for collaterals, which were graded according to Rentrop's classification (0 to 3), and a collateral index was calculated (collateral sum divided by 3). Preoperative and postoperative radionuclide ventriculograms provided global LVEF and regional ejection fractions. Of 123 regions evaluated, 120 were dysfunctional at baseline. Virtually all (122 of 123) regions were subtended by an artery with > or = 70% stenosis that was bypassed. Thirty-eight (81%) of 47 dysfunctional regions with grade 2 or 3 collaterals improved regional ejection fraction after surgery versus 38 (52%) of 73 dysfunctional regions with grade 0 or 1 collaterals (P = 0.0018). Global LVEF was 34 +/- 10% after surgery (P < 0.001 versus before surgery). Among patients with a global LVEF increase > or = 10%, collateral index was 1.81 versus 0.83 in those with an LVEF increase < 10% (P = 0.005). CONCLUSIONS: In this population of patients with coronary artery disease with severe LVD, the presence of angiographic grade 2 or 3 collaterals predicted recovery of regional and global myocardial function after CABG. PMID- 9852880 TI - Delay in revascularization is associated with increased mortality rate in patients with severe left ventricular dysfunction and viable myocardium on fluorine 18-fluorodeoxyglucose positron emission tomography imaging. AB - BACKGROUND: The identification of high-risk patients who require early revascularization has become increasingly important with the present emphasis on reducing health care resources. This is particularly relevant to health care systems with prolonged waiting times for interventions. Myocardial viability imaging with the use of fluorine 18-fluorodeoxyglucose (FDG) PET may help to identify high-risk patients with severe left ventricular dysfunction. The aim of this study was to evaluate the consequences of prolonged waiting time on cardiac outcomes in patients with left ventricular dysfunction directed to revascularization based on FDG PET imaging. METHODS AND RESULTS: Forty-six patients with coronary disease and an ejection fraction of < or = 35% were considered candidates for revascularization based on FDG PET viability imaging. Thirty-five of 46 patients were subsequently accepted for revascularization. Patients were divided into 2 groups based on the median waiting time after PET: an early group (< 35 days; n = 18) and a late group (> or = 35 days; n = 17). Preoperative mortality rates were significantly increased in the late group (4 of 17 [24%] versus 0 of 18 in the early group; P < 0.05). In postoperative follow-up (17 +/- 7 months), cardiac events occurred in 2 of 18 (11%) and 1 of 13 (7.8%) patients in the early and late groups, respectively. Left ventricular ejection fraction increased after early revascularization (24 +/- 7% to 29 +/- 8%, P < 0.001, baseline versus 3 months) but not in the late group (27 +/- 5% to 28 +/- 6%, P = NS). CONCLUSIONS: Preoperative FDG PET can be used to identify a high risk group of patients who may benefit from early revascularization. A long waiting time for revascularization is associated with a high mortality rate and suggests that early revascularization is desirable after the identification of hibernating viable myocardium. PMID- 9852882 TI - Angiogenesis in transmyocardial laser revascularization. A nonspecific response to injury. AB - BACKGROUND: The mechanism of action of transmyocardial laser revascularization (TMR) is poorly understood. TMR has been shown to stimulate angiogenesis in porcine and canine myocardium. METHODS AND RESULTS: We examined the possibility that angiogenesis also occurs in ovine myocardium and that it is a nonspecific tissue injury response. Five Dorset sheep underwent creation of transmyocardial channels of equal diameter in both the apical and basal regions of the left ventricle through the use of both CO2 laser in 1 region and a power drill in the alternate region of the same heart. All channels were closed at 4 weeks. Histology showed channel remnants composed of granulation tissue, fibrosis, and new vessels (NV). These changes were not distinguishable on the basis of the method of channel creation. The average diameter of the channels was similar (laser, 630 +/- 180 microns; drill, 750 +/- 280 microns) (P = NS). NV with smooth muscle media were seen within the channel remnant and immediately surrounding this region using Verhoeff-Van Gieson (elastic) stain. The densities of the NV within the channel remnants were similar (laser, 1.87 +/- 1.05 NV/high-power field [hpf]; drill, 1.92 +/- 1.09 NV/hpf; P = NS), and both were significantly greater than the density of vessels in remote regions, > 5 mm from the channel center (remote laser area, 0.09 +/- 0.28 NV/hpf; remote drill area, 0.04 +/- 0.21 NV/hpf; P = NS for remote areas, P < 0.001 for laser versus remote laser, P < 0.001 for drill versus remote drill area). CONCLUSIONS: These findings demonstrate that the CO2 laser stimulates angiogenesis in normal ovine myocardium and suggest that this response represents a nonspecific reaction to tissue injury. PMID- 9852883 TI - Nontransmural laser treatment incompletely denervates canine myocardium. AB - BACKGROUND: Clinical experience with transmyocardial laser revascularization (TMLR) has reproducibly demonstrated an improvement in angina class. Denervation has been implicated as a mechanism whereby this clinical effect may be achieved. Because endovascular techniques for TMLR are currently under development, we investigated the impact of nontransmural endoventricular laser treatment on cardiac nerves in a canine model. METHODS AND RESULTS: Five mongrel dogs underwent creation of nontransmural endoventricular channels in the anterior left ventricle with a Holmium:YAG laser. Cardiac afferent nerve function was assessed in control and treatment regions by the epicardial application of bradykinin, a potent algesic, at initial thoracotomy before laser treatment, and at repeat thoracotomy 2 weeks later. The resulting central nervous system-mediated decrease in systemic mean arterial pressure seen in all animals at baseline was reduced by 90% at 2 weeks in the laser-treated territory but was preserved in controls. Immunoblot analysis of tissue samples taken from laser-treated regions demonstrated a 66% reduction in tyrosine hydroxylase, a sympathetic nerve specific enzyme, as assessed by densitometry. Enzyme content was unchanged in control regions. CONCLUSIONS: These data suggest that nontransmural endoventricular laser treatment only partially denervates the heart. This may have implications for the clinical efficacy of the endovascular approach in the relief of angina pectoris. PMID- 9852884 TI - Transmyocardial revascularization in patients with refractory, unstable angina. AB - BACKGROUND: Previous reports of transmyocardial revascularization (TMR) indicate a significant mortality in patients with refractory, unstable angina. We hypothesized that TMR with a holmium laser would result in significant angina relief with acceptable mortality in this patient population. METHODS AND RESULTS: Patients were defined as unstable if they were unweanable from intravenous antianginal medications or were too unstable for a persantine thallium scan. Patients had a left ventricular ejection fraction (LVEF) of > 25% and were not amenable to CABG or PTCA. Before treatment, all patients had class IV angina. TMR was performed in 85 patients, with a mean of 35 +/- 11 transmural laser channels. Mean age was 63 +/- 10 years. Mean LVEF was 48 +/- 11%. Of these patients, 79% were men. Prior CABG and/or PTCA had been performed in 87% of patients, and 72% of patients had a history of prior MI. Operative mortality was 12% (10 of 85). There were 2 deaths between discharge and 3 months after surgery and 7 late deaths from 6 to 12 months after surgery. Twelve-month mortality was 22.4% (19 of 85). At 3 months, 86% of patients had class II angina or better. At 6 and 12 months, 77% and 75% of patients, respectively, had class II angina or better. Mean angina class at 6 and 12 months' follow-up was 1.5 +/- 1.1 and 1.6 +/- 1.3, respectively. CONCLUSIONS: In patients with refractory unstable angina, TMR with a holmium laser provided significant angina relief. Moreover, 30-day operative mortality and 12-month mortality were acceptable, especially given this subset of unstable patients with refractory angina. PMID- 9852886 TI - Quality of life in patients with bioprostheses and mechanical prostheses. Evaluation of cohorts of patients aged 51 to 65 years at implantation. AB - BACKGROUND: The purpose of this study was 3-fold: to compare the quality of life (QOL) in age- and sex-matched patients who received biological and mechanical prosthetic valves in isolated aortic valve replacement, to compare the QOL of patients with aortic valve replacement with the general population, and to compare patients with biological and mechanical prostheses with certain valve specific questions and relate these responses to overall QOL. METHODS AND RESULTS: Patient-perceived QOL was evaluated in 200 patients who were sampled from a population of 420 patients (age range 51 to 65 years) who underwent isolated aortic valve replacement in the period of 1986 to 1996. One hundred of the sampled patients had a mechanical valve inserted and an equal number had porcine bioprostheses. Three survey instruments were used to examine perceived QOL: the SF-12 form, a 7-valve specific question form, and the Lamy Smiley Faces form. The response to the questionnaires was 89.5% (179 patients). Patients with mechanical valves were more bothered by valve sounds (P < 0.01) and had a negative correlation (P < 0.01) between valve sound and QOL on the mental scale only. Patients with biological valves were more fearful of the need for reoperation (P < 0.01), but there was no correlation between fear and QOL. The mechanical valve group had a negative correlation (P < 0.01) between fear of reoperation and QOL on both the mental and physical scales. There was no difference between the 2 cohorts with respect to fear of valve failure. Patients with mechanical valves were more concerned about frequency of medical visits and blood tests (P < 0.01) as well as the possibility of anticoagulant-related bleeding events (P < 0.01). QOL was equivalent between the 2 groups and to the general population for the same age group. Ninety-seven percent of the patients indicated they would make the same surgical decision again with regard to valve replacement; there was no difference between the 2 valve groups on this question. CONCLUSIONS: Patient-perceived QOL is similar between patients with aortic mechanical and biological valve replacement in the studied age group and comparable to the general population of similar age. Although certain valve specific differences exist between the 2 prosthetic types, these differences do not appear to affect overall QOL as described by these patients. PMID- 9852885 TI - Effect of implantable cardioverter-defibrillator implantation on surgical morbidity in the CABG Patch Trial. Surgical Investigators of the Coronary Artery Bypass Graft Patch Trial. AB - BACKGROUND: The Coronary Artery Bypass Graft (CABG) Patch Trial tested the hypothesis that prophylactic insertion of an implantable cardioverter defibrillator (ICD) improves survival rates after high-risk CABG. We compared group-specific perioperative morbidity and mortality rates. METHODS AND RESULTS: Patients were randomized intraoperatively to undergo CABG (control subjects, n = 454) or CABG plus ICD implantation (n = 446). There were no significant differences between groups in the incidence of diabetes, ejection fraction < 0.25, end-diastolic pressure, prior myocardial infarction, or congestive heart failure. Cardiopulmonary bypass time averaged 106 minutes in control subjects and 127 minutes in the ICD group. At the inception of the trial, investigators were concerned that ICD therapy could increase surgical mortality rates or the incidence of shock, bleeding, congestive heart failure, arrhythmias, or deep sternal wound infection. Of these, only sternal wound infection was significantly more frequent in the ICD group (2.2% versus 0.4%, P < 0.05). Also more common in the ICD group were infection at a wound or catheter site (12% versus 6%), urinary tract infection (4% versus 1%), pneumonitis (8% versus 4%), respiratory insufficiency (13% versus 8%), transient central nervous system deficit (6% versus 2%), and psychotic reaction (4% versus 1%). The all-cause death rate was 6.7% in the ICD group and 4.6% for control patients (P = NS) at the time of the last surgical death, postoperative day 48. CONCLUSIONS: Epicardial ICD insertion during CABG is associated with an increase in perioperative infection. Although reporting bias may have influenced the data, if ICD insertion is indicated in CABG patients, metachronous endocardial implantation should be considered. PMID- 9852887 TI - Twenty-year, three-institution evaluation of the Hancock Modified Orifice aortic valve durability. Comparison of actual and actuarial estimates. AB - BACKGROUND: Information regarding the incidence of structural valve deterioration (SVD) is used in selecting the type of valve for patients. Standard actuarial statistical techniques have been used widely but do not provide the most appropriate information for patient populations experiencing competing hazards. "Actual," or cumulative incidence, methods may provide a better estimate of the durability of tissue valves for these patients. The purpose of this article is to compare actuarial and actual estimates of the durability of the Hancock Modified Orifice bioprosthesis aortic valve in a multi-institutional study. METHODS AND RESULTS: Valves were implanted between 1976 and 1985 in 3 institutions. This sample contains follow-up data on 727 patients (42% female) with a mean age of 63 +/- 13 years. The difference between actuarial and actual rates of SVD became more important over time. At 5 years, the difference is significant only in the elderly (aged > or = 65 and > or = 70), whereas at 17 years, the difference was significant for all patients. Similarly, the magnitude of the difference increases over time. Freedom from SVD for patients > 65 at 5 years is estimated at 98 +/- 0.01% by actuarial methods and 100 +/- 0.00% actual methods. The difference between estimates is larger at 10 years, 93 +/- 0.02% versus 96 +/- 0.01%. This difference is greater at 17 years, 78 +/- 0.04% versus 93 +/- 0.01%. CONCLUSIONS: The results of this study demonstrate that the particular statistical analysis method used to calculate SVD can provide strikingly different conclusions. These observations indicate that the actual method is able to identify the lower risk of SVD in older patients. Generalizability issues must be considered, however, when using actual SVD rates to make decisions regarding valve selection in larger populations. PMID- 9852888 TI - Anticoagulation is unnecessary after biological aortic valve replacement. AB - BACKGROUND: Opinion differs as to whether anticoagulation is beneficial in preventing ischemic stroke in the early postoperative period after biological aortic valve replacement (AVR). The purpose of this study was to determine whether early anticoagulation with heparin and warfarin confers any significant advantage for patients undergoing such replacement. METHODS AND RESULTS: Patients undergoing biological AVR between 1987 and 1996 were divided retrospectively into 2 groups based on their postoperative anticoagulation. Group A (109 patients) received heparin followed by warfarin for 3 months (prothrombin time, 20 to 25 seconds). Group B (76 patients) received no postoperative anticoagulation. Patients were followed for cerebral ischemic events, bleeding, repeat operation, hospital stay, and survival. There were 5 (4.6%), 3 (2.8%), and 12 (11%) postoperative cerebral ischemic events for group A at time points of < 24 hours, 24 hours to 3 months, and > 3 months, respectively; for group B patients, 3 (3.9%), 2 (2.6%), and 9 (11.8%) events were seen during the same respective time periods. There were no statistically significant differences for ischemic events during any of these time periods for the 2 groups. Bleeding complications occurred in 10 (9.2%) group A and 7 (9.2%) group B patients. Mean hospital stay was 12 days for both groups. Repeat operative AVR was required in 6 (5.5%) group A and 7 (9.2%) group B patients. A comparison of Kaplan-Meier survival rates between groups A and B (mean follow-up, 47 +/- 26 and 59 +/- 30 months, for groups A and B, respectively) was not statistically significant (P = 0.60). Survival rates were 93%, 84%, and 62% at 1, 5, and 7 years for group A and 87%, 74%, and 67% for group B, respectively. CONCLUSIONS: Early anticoagulation after AVR confers no advantage in the prevention of early cerebral ischemic events after biological AVR. No disadvantage in terms of bleeding or prolonged hospital stay was incurred by early anticoagulation. Long-term valve function and survival were not adversely affected by withholding early anticoagulation. We conclude that early anticoagulation after biological AVR is unnecessary. PMID- 9852889 TI - Valve replacement in chronic aortic regurgitation. True predictors of survival after extended follow-up. AB - BACKGROUND: The influence of preoperative clinic, hemodynamic, and surgical procedures on extended long-term prognosis after valve replacement for chronic aortic regurgitation is still unclear. METHODS AND RESULTS: One hundred ninety two patients (mean age, 44 years) who underwent valve replacement for chronic aortic regurgitation between 1970 and 1983 were followed for 13 to 26 years (mean follow-up, 13.8 years; surviving patients, 18.7 years). Perioperative mortality was 2.6%; survival rates after 10 and 20 years were 76% and 55%; 65 of 80 deaths were cardiac; 21% of patients were free of cardiac complications after 20 years; and 83 of 100 long-term survivors were in NYHA classes I and II. In a univariate analysis, age at operation (P < 0.0001), higher preoperative NYHA class (P < 0.0001), lower left ventricular (LV) ejection fraction (P = 0.0001), higher end systolic volume (P = 0.0007), history of endocarditis (P = 0.0004), and additional surgical intervention besides valve replacement (P = 0.004) were the main predictors of late survival. In a multivariate stepwise Cox analysis, age (P = 0.0004), high LV end-systolic volume (P = 0.0004), higher NYHA class (P = 0.01), and previous endocarditis (P = 0.006) were independent predictors. CONCLUSIONS: In chronic aortic regurgitation, symptoms and LV systolic function are the main predictors for extended long-term outcome after valve replacement. Low operative mortality and good extended survival make valve replacement mandatory in mildly or asymptomatic patients when LV systolic function decreases and considerable enlargement of systolic dimensions occur. PMID- 9852890 TI - Excess mortality due to coronary artery disease after valve surgery. Secular trends in valvular regurgitation and effect of internal mammary artery bypass. AB - BACKGROUND: During the 1980s, mortality from coronary artery disease (CAD) decreased markedly in the United States. This raises the question of whether a parallel decrease occurred in excess mortality due to CAD in patients undergoing surgical correction of valvular regurgitation. METHODS AND RESULTS: Survival of 752 patients (age, 64 +/- 13 years) with isolated left-sided valvular regurgitation operated on from 1980 to 1991 was analyzed. Of 242 patients with CAD (stenosis > or = 70%), 208 had coronary artery bypass grafting. Multivariate analysis identified CAD as an independent predictor of operative mortality (odds ratio [OR] = 2.35, P = 0.012), overall (hazard ratio [HR] = 1.65, P < 0.0001) and late mortality (HR = 1.57, P = 0.0006), and postoperative congestive heart failure (HR = 2.35, P = 0.0001). Comparison of patients operated on in 1980 to 1985 with those operated on in 1986 to 1991, excess of operative, overall, and late mortality and postoperative congestive heart failure (adjusted for age and gender) related to associated CAD did not decrease significantly (P = 0.23, P = 0.64, P = 0.90, and P = 0.61, respectively). Overall survival was better for patients receiving an internal mammary artery graft than those receiving vein grafts only (HR = 0.57, P = 0.011). CONCLUSIONS: In contrast to the secular trend for decreased mortality from CAD, excess mortality related to associated CAD after surgery for valvular regurgitation has not decreased. Internal mammary artery grafts were associated with improved outcome. In patients with valvular regurgitations, these results support continued active search of associated CAD, wide use of internal mammary artery graft, and vigorous efforts for secondary prevention of complications of CAD. PMID- 9852891 TI - Choice of mitral prosthesis in the elderly. An analysis of actual outcome. AB - BACKGROUND: In younger patients requiring mitral valve replacement (MVR), mechanical prostheses (MPs) have been reported to give better freedom from all valve-related complications (VRCs) because of the high incidence of late valve degeneration (VD) associated with bioprostheses (BPs). In older patients, however, the risk of VD may be reduced because of the large competing risk of noncardiac death (NCD). Previous studies on VD in the elderly have used actuarial analysis, which overestimates the risk of VD in this population because it assumes that dead patients are still at risk. In contrast, cumulative incidence (actual) analysis acknowledges that patients who die have no risk of VD. This study compares the results of both "actual" and "actuarial" analyses of the freedom from VD in elderly patients undergoing MVR. METHODS AND RESULTS: From June 1976 through January 1996, 504 patients > or = 70 years of age underwent MVR at our institution. Isolated mitral operations were performed in 159 patients, and 169 had concomitant CABG. Hospital mortality was 59 of 374 (15.9%) for tissue prosthesis versus 24 of 130 (18.5%) for mechanical prosthesis (P = NS). For tissue versus mechanical prosthesis, 10-year freedom from noncardiac death was 75.0% versus 67.6% (P = NS); 10-year actuarial freedom from valve degeneration was 79.8% versus 93.4% (P = NS); 10-year actual freedom from valve degeneration was 92.6% versus 95.4% (P = NS); and 10-year actual freedom from all VRCs was 84.4% versus 92.3% (P = NS). CONCLUSIONS: In elderly patients undergoing MVR, actuarial analysis overestimates the 10-year risk of VD compared with actual analysis (20.2% versus 7.4% for BP, 6.6% versus 4.6% for MP). In these patients, the actual freedoms from VD and all VRCs do not differ significantly between BP and MP. Thus, in this age group, the necessity for anticoagulation or its avoidance may be the predominant factor in choosing a replacement mitral valve. PMID- 9852892 TI - Comparison of open mitral commissurotomy with mitral valve replacement with or without chordal preservation in patients with mitral stenosis. AB - BACKGROUND: Percutaneous balloon mitral valvuloplasty and durable mitral prostheses have made the role of open mitral commissurotomy (OMC) uncertain. METHODS AND RESULTS: Results from the use of St Jude mitral valve replacement (SJMVR) were compared with those of the use of OMC in 312 consecutive patients with mitral stenosis between 1983 and the present. OMC and SJMVR patients were well matched for age, sex, and comorbidity except that SJMVR patients had more severe stenosis and were more likely to undergo concurrent aortic valve replacement. Compared with OMC, SJMVR without chordal preservation involved a longer pump time (158 +/- 81 versus 87 +/- 41 min, P < 0.05), more frequent in hospital complications or death (57 of 219 [26%] versus 4 of 52 [8%], P < 0.01), and longer hospital stay (13 +/- 11 versus 10 +/- 6 days, P = 0.001). Preservation of chordae to at least 1 mitral valve leaflet decreased early morbidity and mortality rates of SJMVR to values comparable to those of OMC (3 of 41 [7%]). Survival was greater at 10 years for OMC versus SJMVR (86 +/- 5% versus 67 +/- 4%, P = 0.03). Ten-year freedom from cardiac events was not different between groups (49 +/- 9% for OMC versus 55 +/- 4% for SJMVR, P = 0.7). Freedom from subsequent mitral procedures at 10 years was better for SJMVR (96 +/- 2% versus 58 +/- 8%, P < 0.001). CONCLUSIONS: In the modern era, SJMVR offers significantly greater durability than does OMC. Chordal preservation at the time of SJMVR may reduce perioperative complications to levels comparable to those of OMC. PMID- 9852893 TI - Mitral valve repair in cardiomyopathy. AB - BACKGROUND: Volume overload secondary to mitral regurgitation (MR) in cardiomyopathy is considered critical in the pathogenesis of subsequent ventricular dysfunction. Open mitral valve repair (OMVP) is hypothesized to improve symptomatology and ventricular function by reducing the volume overload of the left ventricle. METHODS AND RESULTS: All patients who underwent OMVP with a left ventricular ejection fraction (EF) of < 0.30 (n = 81) from 1984 through 1997 were reviewed (1 patient was lost to follow-up). Fifteen operations (18.5%) were repeat operations after previous coronary artery bypass graft surgery. Preoperative and postoperative EFs and NYHA class were compared. Survival probabilities were calculated, and multivariate analysis was performed. The average age of all patients was 67.1 years (range, 41 to 83 years). Mean follow up was 1.7 years (range, 2 months to 8.5 years). The most common mitral repair was ring annuloplasty. Sixty-two patients (77%) had concomitant coronary artery bypass graft surgery. The surgery mortality rate was 11% (9 of 81); 6 of these 9 patients were > 70 years old. The overall Kaplan-Meier survival probability rate at 1, 2, 3, 4, and 5 years was 0.73, 0.68, 0.58, 0.50, and 0.38, respectively. EF improved significantly (0.24 to 0.32; P < 0.0001), as did the NYHA class (3.2 to 1.6; P < 0.0001), at follow-up. There was no difference in late survival between patients with an EF of < 0.20 (21 patients) and those with an EF between 0.20 and 0.30 (P = NS). Risk factors for death included heart failure and old age. CONCLUSIONS: OMVP for MR in the setting of ischemic cardiomyopathy and low EF appear to improve ventricular function, medium-term patient symptomatology, and survival. PMID- 9852894 TI - Semirigid or flexible mitral annuloplasty rings do not affect global or basal regional left ventricular systolic function. AB - BACKGROUND: Previous studies have revealed that rigid mitral annuloplasty rings may be associated with left ventricular (LV) systolic dysfunction, but whether ring type affects regional systolic function at the base of the LV, in the region near the mitral annulus, is unclear. We tested the hypothesis that rigid fixation of the mitral annulus results in significant regional systolic dysfunction at the base of the LV. METHODS AND RESULTS: Twenty-six adult male sheep underwent placement of 13 miniature tantalum markers into the LV epicardium and around the mitral annulus to allow calculation of LV volume and regional epicardial area. Group I (n = 7) sheep served as controls; animals randomized to groups II (n = 11) and III (n = 8) underwent mitral annuloplasty with either a semirigid or flexible ring, respectively. After a 7- to 10-day recovery period, animals were studied in a closed-chest, sedated, autonomically blocked state. Global LV systolic function (end-systolic elastance and preload recruitable stroke work) were not significantly different among the 3 groups (P = 1.0, ANOVA). Regional systolic function at the base of the LV (fractional area shrinkage [FAS] of 4 epicardial areas) at comparable LV preload and afterload was similar in the 4 basal areas (P = 0.223, MANOVA). With the use of load-insensitive indexes (slope and area intercept of the end-systolic pressure-regional area relationship and regional stroke work-end-diastolic area relationship), regional systolic function also was not different between groups at baseline or with inotropic stimulation in any basal region (P > 0.05, MANOVA). Furthermore, neither annuloplasty ring perturbed the regional pattern of basal LV systolic function. CONCLUSIONS: Postoperative LV systolic function, both globally and in the region of the base of the LV (near the mitral annulus), was not altered with either semirigid or flexible ring fixation of the mitral annulus. PMID- 9852895 TI - Decreasing incidence of stroke during valvular surgery. AB - BACKGROUND: The predictors and causes of stroke after valvular surgery are incompletely defined. We examined the incidence, predictors, and mechanisms of stroke during valvular procedures over a 15-year time period. METHODS AND RESULTS: We retrospectively reviewed prospectively gathered data on 5954 consecutive patients undergoing valvular procedures at our institution from 1982 to 1996. Stroke was defined as persistent central nervous system deficit, usually with confirmatory CT imaging. Patients were divided into 3 groups according to date of operation: group 1, 1982 to 1986 (n = 1819); group 2, 1987 to 1991 (n = 2022); and group 3, 1992 to 1996 (n = 2113). Chart review was undertaken of all patients who developed stroke (n = 189). Stroke occurred in 3.8% of group 1 patients, 3.3% of group 2, and 2.6% of group 3 (P = 0.120). The decreasing incidence of stroke over time was confirmed by multivariable logistic regression analysis, in which earlier date of operation was an independent risk factor for stroke (P < 0.001). Predictors of stroke identified by multivariable logistic regression were (listed in decreasing order): (1) endocarditis (OR, 3.0; 95% CI, 1.8 to 5.0); (2) age > 74 years (OR, 2.3; 95% CI, 1.5 to 3.7); (3) earlier time period of operation (1982 to 1986: OR, 2.2; 95% CI, 1.5 to 3.2; 1987 to 1991: OR, 1.5; 95% CI, 1.0 to 2.2); (4) urgent timing (OR, 2.0; 95% CI, 1.4 to 2.8); (5) concomitant coronary bypass (OR, 2.0; 95% CI, 1.4 to 2.8); and (6) reoperation (OR, 1.7; 95% CI, 1.2 to 2.4). In more recent years of operation, we found an increasing prevalence of age > 74 years (7.4% in group 1, 9.5% in group 2, and 15.3% in group 3; P < 0.001), urgent timing (11%, 26%, and 34%, P < 0.001), and concomitant coronary bypass surgery (25%, 27%, and 33%; P < 0.001). CONCLUSIONS: The incidence of stroke during valvular surgery has decreased with time, despite an increased prevalence of risk factors. Predictors of stroke suggest 3 major causes (multivariable predictors in parentheses): atherosclerotic emboli (elderly age, concomitant coronary bypass), shock (urgent timing, reoperation), and septic emboli (endocarditis). PMID- 9852896 TI - Elevated insulin-like growth factor-I and transforming growth factor-beta 1 and their receptors in patients with idiopathic hypertrophic obstructive cardiomyopathy. A possible mechanism. AB - BACKGROUND: Idiopathic hypertrophic obstructive cardiomyopathy (HOCM) is characterized by regional myocardial hypertrophy. In our previous study, we demonstrated that mRNA levels for insulin-like growth factor-I (IGF-I) and transforming growth factor-beta 1 (TGF-beta 1) were elevated in HOCM tissue. In this study, we investigated IGF-I and TGF-beta 1 protein levels and their respective receptor levels and localization. METHODS AND RESULTS: Myocardial growth factor protein levels were quantified with the use of chemiluminescent slot blot analysis with monoclonal antibodies against IGF-I and TGF-beta. The growth factor receptor binding sites were evaluated with 125I-labeled IGF-I and TGF-beta 1. The receptors were localized with immunohistochemistry. Data were expressed as mean +/- SEM. IGF-I and TGF-beta protein levels in HOCM myocardium (351.8 +/- 46.5 and 17.4 +/- 2.0 ng/g tissue, respectively; n = 6) were significantly higher (P < 0.01 for all groups) than in non-HOCM myocardium obtained from patients with aortic stenosis (AS, 182.1 +/- 22.7 and 8.0 +/- 1.2 ng/g tissue, respectively; n = 5), stable angina (SA, 117.4 +/- 20.9 and 7.5 +/- 2.7 ng/g tissue, respectively; n = 5), and transplanted hearts (TM, 166.3 +/- 30.1 and 6.4 +/- 1.2 ng/g tissue, respectively; n = 5). Maximal and high-affinity binding sites for IGF-I receptor in the HOCM were greater (P < 0.01 and P < 0.05) than the levels in AS, SA, and TM. The maximal receptor binding sites for TGF beta 1 in HOCM were greater (P < 0.05) than those for SA and TM. Immunohistochemistry demonstrated that IGF-I and TGF-beta 1 receptors were located on the cardiomyocytes and TGF-beta 1 receptors were located on the fibroblasts. CONCLUSIONS: Increased IGF-I and TGF-beta 1 gene expression previously observed in HOCM myocardium results in elevated protein levels. IGF-I and TGF-beta 1 signals may be further amplified by increased receptor numbers on cardiomyocytes and fibroblasts. The data suggest a possible autocrine mechanism of IGF-I-stimulated cardiomyocyte hypertrophy and a paracrine mechanism of TGF beta 1-stimulated extracellular matrix overproduction in HOCM. PMID- 9852897 TI - Can cellular transplantation improve function in doxorubicin-induced heart failure? AB - BACKGROUND: Transplantation of fetal cardiomyocytes has been shown to improve function of regionally infarcted myocardium, but its effects on global heart failure are still unknown. METHODS AND RESULTS: Heart failure was induced in female mice by intraperitoneal injection of doxorubicin (2 mg/kg twice per week over 2 cycles of 2 weeks separated by a 2-week drug-free period). One week after the end of treatment, left ventricular function was assessed by transthoracic echocardiography (baseline). Animals were then randomized into 3 groups: The treated group (n = 12) received an intramyocardial injection of fetal cardiomyocytes (1 x 10(6) in 10 microL) harvested from transgenic mice expressing the gene of beta-galactosidase, the control group (n = 15) received an equivalent volume of culture medium alone, and 10 sham mice had no surgery. Two weeks and 1 month after transplantation, function was again assessed echocardiographically. At baseline, fractional shortening was not significantly different between the 3 groups. It then significantly increased in cell-treated mice at 2 weeks and 1 month after transplantation (P < 0.002 and P < 0.03 versus baseline, respectively), whereas it did not change in untreated animals. Transplanted cells could not be identified by beta-galactosidase activity or presence of Y chromosome (with 1 exception). CONCLUSIONS: Cellular transplantation can improve function of globally failing hearts by a mechanism that might not necessarily involve the sustained presence of transplanted cells but rather the effects of cardioprotective factors released by them. PMID- 9852898 TI - Impact of University of Wisconsin solution on clinical heart transplantation. A comparison with Stanford solution for extended preservation. AB - BACKGROUND: The University of Wisconsin Solution (UW) has extended preservation of abdominal organs but has not allowed equally extended preservation of the heart. Therefore, the impact of UW on clinical heart transplantation has remained unclear. METHODS AND RESULTS: Between June 1986 and March 1994, 161 orthotopic heart transplants were performed at our center. Of these, 66 were preserved for > or = 3 hours. Of these, 17 hearts were preserved with Stanford solution (STNF), which was used before 1990, and 49 were preserved with UW. These groups were compared for indexes of ischemic injury, ventricular function, and survival. The UW group contained more status-1 recipients (57% versus 29%, P < 0.05) and a higher mean donor age (30.7 versus 22.1 years, P = 0.008). Mean ischemic time was slightly but not significantly higher with UW (228 versus 205 minutes for UW versus STNF, respectively; P = 0.085). The time to wean from bypass after cross clamp removal was nearly twice as long with STNF than with UW (80.6 versus 44.3 minutes, P < 0.001). There was no difference in the incidence of primary graft failure (2% for UW versus 6% for STNF, P = 0.43). The average need for inotropic support over the first 8 posttransplant hours was significantly higher with STNF than UW. Neither hospital stay nor survival differed. Nevertheless, the ability to use donor organs from more distant sites was increased. Of all hearts preserved with STNF, 26% were stored for > or = 3 hours, whereas 51% of all hearts preserved in UW were stored for this length of time. Donor use of hearts increased from 20% in 1989 to 63% in 1993, largely because of greater use of more distant donors. CONCLUSIONS: We conclude that heart preservation with UW limits ischemic damage from prolonged storage and improves myocardial function in the early posttransplant period, thus allowing greater use of available donors from distant sites to patients awaiting heart transplantation. PMID- 9852899 TI - Tacrolimus-based partial conditioning produces stable mixed lymphohematopoietic chimerism and tolerance for cardiac allografts. AB - BACKGROUND: Thoracic organ transplantation remains limited by the reciprocal problems of rejection and the toxicities of nonspecific immunosuppression. Mixed bone marrow chimerism reliably produces donor-specific transplantation tolerance without immunosuppressive drugs. We have previously described a nonmyeloablative conditioning regimen based on recipient treatment with antilymphocyte serum, tacrolimus, and low-dose total-body irradiation that yields long-term multilineage allogeneic bone marrow chimerism in the rat. We have now investigated whether mixed bone marrow chimerism that arises from this partial conditioning strategy produces permanent acceptance of donor-specific cardiac allografts. METHODS AND RESULTS: Mixed allogeneic chimeras (ACI-->WF) were prepared by treating Wistar Furth recipients with a single dose of antilymphocyte serum 5 days before bone marrow transplantation and tacrolimus 1 mg/kg/d from days -1 to 10. Five hundred cGy total-body irradiation was administered immediately before infusion of 1 x 10(8) donor (ACI) T-cell depleted marrow cells. All recipients were chimeric, with a mean level of donor chimerism = 26.3 +/- 3.5%. Chimeras underwent heterotopic cardiac transplantation 4 weeks after bone marrow transplantation. All donor-specific (ACI) grafts were permanently accepted (follow-up, 230 to 360 days). Third-party grafts were rapidly rejected. Histology of long-surviving donor-specific grafts was without evidence of acute or chronic rejection. Second-set donor-specific skin grafts transplanted to chimeras 135 days after heart transplantation showed long-term survival (> 130 days), whereas third-party skin grafts were rapidly rejected. Mixed lymphocyte reaction demonstrated in vitro donor-specific hyporeactivity. CONCLUSIONS: A tacrolimus-based nonmyeloablative recipient conditioning regimen produces mixed bone marrow chimerism and donor-specific tolerance to cardiac allografts in the rat. PMID- 9852900 TI - Minimum hematocrit at differing cardiopulmonary bypass temperatures in dogs. AB - BACKGROUND: The purpose of this study was to determine the minimum hematocrit (Hct) supporting cerebral oxygenation over the range clinically relevant cardiopulmonary bypass (CPB) temperatures in dogs. The effect of hemodilution on cerebral blood flow (CBF), cerebral metabolic rate (CMRO2), and cerebral oxygen delivery (CDO2) was determined over a range of Hcts during CPB at 38 degrees C, 28 degrees C, and 18 degrees C. METHODS AND RESULTS: Measurements were obtained at target CPB temperature and after progressive normovolemic hemodilution in 3 groups of 8 anesthetized animals. Dextran 70 (6%) was used as a diluent. CBF was measured by use of the sagittal sinus outflow technique. CMRO2 and CDO2 were calculated by the use of standard formulae. In each temperature group, hemodilution was associated with a reciprocal rise in CBF. As Hct was reduced to 0.10 +/- 0.02 at 38 degrees C, 28 degrees C, and 18 degrees C, CBF increased 260%, 220%, and 160% of the control nonhemodiluted value. Increases in CBF helped compensate for decreased arterial oxygen content and maintain CDO2. With progressive temperature reduction, these compensatory flow increases were reduced and CDO2 was decreased at lesser degrees of hemodilution. Statistical analysis indicated that cerebral oxygen demand was maintained to an Hct of 0.14, 0.11, and < or = 0.10 in the 38 degrees C, 28 degrees C, and 18 degrees C groups; however, physiologically important changes in cerebral oxygen supply occur at Hcts of approximately 0.18, 0.15, and 0.12, respectively, at those temperatures. CONCLUSIONS: This investigation systematically characterizes the critical Hct supporting cerebral oxygenation at differing CPB temperatures. Over a range of CPB temperatures, the curve describing the relationship between Hct and cerebral oxygen balance has a broad plateau, indicating cerebral tolerance for a wide range of Hcts. The minimum Hct that supports cerebral oxygenation is shifted leftward as temperature is reduced, but the reduction in critical Hct is not proportional to the reduction in CMRO2. Although we do not advocate hemodilution to these extreme values, we find that these data provide a physiological foundation for our hemodilution practice and provide some guidance for management of Hct as body temperature changes during CPB. PMID- 9852901 TI - ATP-sensitive potassium channel activation before cardioplegia. Effects on ventricular and myocyte function. AB - BACKGROUND: Pretreatment with potassium channel openers (PCOs) has been shown to provide protective effects in the setting of myocardial ischemia. The goal of the present study was to examine whether PCO pretreatment would provide protective effects on left ventricular (LV) and myocyte function after cardioplegic arrest. METHODS AND RESULTS: The first study quantified the effects of PCO pretreatment on LV myocyte contractility after simulated cardioplegic arrest. LV porcine myocytes were randomly assigned to 3 groups: (1) normothermic control: 37 degrees C x 2 hours (n = 116); (2) cardioplegia: K+ 24 mEq/L, 4 degrees C x 2 hours followed by reperfusion and rewarming (n = 62); and (3) PCO/cardioplegia: 5 minutes of PCO treatment (50 mumol/L, SR47063, 37 degrees C; n = 94) followed by cardioplegic arrest and rewarming. Myocyte contractility was measured after rewarming by videomicroscopy. The second study determined whether the effects of PCO pretreatment could be translated to an in vivo model of cardioplegic arrest. Pigs (weight 30 to 35 kg) were assigned to the following: (1) cardioplegia: institution of cardiopulmonary bypass (CPB) and cardioplegic arrest (K+ 24 mEq/L, 4 degrees C x 2 hours) followed by reperfusion and rewarming (n = 8); and (2) PCO/cardioplegia: institution of CPB, antegrade myocardial PCO perfusion without recirculation (500 mL of 50 mumol/L, SR47063, 37 degrees C), followed by cardioplegic arrest (n = 6). LV function was examined at baseline (pre-CPB) and at 0 to 30 minutes after separation from CPB by use of the preload-recruitable stroke work relation (PRSWR; x 10(5) dyne.cm/mm Hg). LV myocyte velocity of shortening was reduced after cardioplegic arrest and rewarming compared with normothermic control (37 +/- 3 vs 69 +/- 3 microns/s, P < 0.05) and was improved with 5 minutes of PCO treatment (58 +/- 3 microns/s). In the intact experiments, the slope of the PRSWR was depressed in the cardioplegia group compared with baseline with separation from CPB (1.07 +/- 0.15 vs 2.57 +/- 0.11, P < 0.05) and remained reduced for up to 30 minutes after CPB. In the PCO-pretreated animals, the PRSWR was higher after cessation of CPB when compared with the untreated cardioplegia group (1.72 +/- 0.07, P < 0.05). However, in the PCO pretreatment group, 50% developed refractory ventricular fibrillation by 5 minutes after CPB, which prevented further study. CONCLUSIONS: PCO pretreatment improved LV myocyte contractile function in an in vitro system of cardioplegic arrest. The in vivo translation of this improvement in contractile performance with PCO pretreatment was confounded by refractory arrhythmogenesis. Thus the application of PCO pretreatment as a protective strategy in the setting of cardiac surgery may be problematic. PMID- 9852902 TI - Optimal myocardial preconditioning in a human model of ischemia and reperfusion. AB - BACKGROUND: Adenosine (ADE) may mediate the protective effects of preconditioning (PC). However, human data are lacking, and the optimal method of ADE administration and the mechanism of protection remain unresolved. METHODS AND RESULTS: We have developed a model of simulated "ischemia" (I) and "reperfusion" (R) in quiescent human ventricular cardiomyocytes. Cellular injury and metabolic parameters were assessed after various interventions: Cells were preconditioned with anoxia (PC0), hypoxia (PC16), anoxic supernatant (SUP0), or hypoxic supernatant (SUP16) with or without the ADE receptor antagonist (SPT) or ADE deaminase (ADA). ADE was applied before, during, or after I or continuously with and without SPT. Cells were treated with the PKC agonist PMA. PC cells were incubated with the protein kinase-C (PKC) antagonist Calphostin-C (Cal-C). PKC translocation and PKC activity were assessed. PC0 was most protective. Protection was transferable via SUP0, which produced the highest concentrations of ADE. Protection was lost with SPT or ADA. Intracellular ATP fell after PC and prolonged I and R. Exogenous ADE was most protective when administered before I at 50 mumol. ADE during I was partially protective. No additional protection was provided with continuous ADE treatment. ADE prevented ATP degradation but increased lactate immediately after its administration. SPT abolished the protective effects of ADE. PMA conferred protection, which was abolished with Cal C. ADE stimulated PKC translocation and PKC activity in the absence of SPT. CONCLUSIONS: Maximal I confers maximal PC. The degree of I is reflected in supernatant ADE concentrations. The initial ATP fall with PC may account for a lack of ATP preservation after I and R. ADE reproduces the protective effects of PC, preserves ATP, and increases lactate production, perhaps by stimulating glycolysis. Clinical trials of ADE administered during cardiac surgery are necessary to further define its beneficial effects in humans. PMID- 9852903 TI - Effects of ischemic preconditioning on myocardial perfusion, function, and microvascular regulation. AB - BACKGROUND: Ischemic preconditioning (PC) has been advocated as a method to preserve myocardial function and perfusion during minimally invasive direct coronary bypass (MIDCAB). We examined the effects of PC on indexes of myocardial function, perfusion, and endothelial and beta-adrenergic coronary regulation after 30 minutes of ischemia and 60 minutes of reperfusion (IR). METHODS AND RESULTS: Five groups of pigs were studied: (1) PC-IR: PC by 3 cycles of 5-minute left anterior descending coronary artery occlusion (CO) and 5-minute reperfusion (Rep) + 30 minutes of CO + 60 minutes of Rep; (2) IR alone: 30 minutes of CO + 60 minutes of Rep; (3) PC alone; (4) PC-IR-glibenclamide (GLIB): PC-IR + infusion of GLIB; (5) control: noninstrumented. Reactivity (in vitro) of coronary arterioles (70 to 150 microns) from the myocardial area at risk was examined with video microscopy. beta-Adrenergic microvascular relaxations to isoproterenol, forskolin, and 8-bromo-cAMP were significantly reduced after IR alone (P < 0.05 versus control, 2-way ANOVA). PC before IR restored these responses to normal (P < 0.05 PC-IR versus IR alone), and GLIB abolished this effect of PC. Subepicardial endothelium-dependent microvascular relaxation to ADP was significantly reduced after IR alone (P < 0.01 versus control) but was preserved in both the PC-IR and PC-IR-GLIB groups (P < 0.05 versus IR alone). The response of vessels to ADP from the subendocardium was significantly reduced in all groups compared with the control response (all P < 0.05 versus control). Nitroprusside elicited a similar response in vessels from all groups. PC before IR did not affect the reduced myocardial percent segmental shortening or left ventricular maximal dP/dt, did not affect myocardial perfusion in the subepicardium or subendocardium, and did not change expression of the inducible or the constitutively expressed isoforms of nitric oxide synthase. CONCLUSIONS: PC before IR preserves beta-adrenergic signal transduction in coronary smooth muscle through a KATP channel mechanism, whereas PC preserves endothelium-dependent relaxation in the subepicardium through a mechanism not related to KATP channels or the enhanced expression of nitric oxide synthase. Nevertheless, PC does not improve short-term myocardial function or baseline myocardial perfusion after IR. Thus, the short-term beneficial role of PC in myocardial protection during MIDCAB may be limited. PMID- 9852904 TI - Ischemic preconditioning in immature myocardium. AB - BACKGROUND: Protection by ischemic preconditioning (PC) has not been studied extensively in immature hearts. We studied the developmental differences in the ability of the rat heart to precondition with ischemia. METHODS AND RESULTS: Hearts from 4-, 7-, 14-, and 21-day-old and adult (approximately 50-day-old) rats were aerobically perfused in Langendorff mode before a PC stimulus of either (1) 3-minute ischemia (I), 3-minute reperfusion (R), 5-minute I, 5-minute R, or (2) 4 cycles of 5-minute I and 5-minute R, before a prolonged I (chosen to give 30% to 40% recovery) and 60-minute R. LVDP recovery was expressed as percent of baseline reading (after 20-minute aerobic perfusion). Protection was seen after protocol 1 in 14- and 21-day-old and adult hearts (45 +/- 5%, 53 +/- 7%, and 58 +/- 5% versus 30 +/- 4%, 29 +/- 3%, and 32 +/- 2% in controls, respectively) but not in 4- and 7-day (neonatal) hearts; neonatal hearts were also not protected in protocol 2. To determine whether this inability of neonatal hearts to precondition was due to insufficient duration of the PC cycle, they were subjected to increased I durations up to 20 minutes before 5-minute R, prolonged I (90 minutes and 60-minute R) (protocol 3); protection was not seen. To determine whether the inability to precondition was due to an excessively prolonged ischemic duration, neonatal hearts were subjected to only 45 minutes of prolonged I (protocol 4); again, PC protection was not evident. CONCLUSIONS: Protection by PC develops after 7 days; the inability of neonatal hearts (< 7 days old) to precondition is not due to insufficient stimulus or extended ischemia. PMID- 9852905 TI - Ischemic preconditioning decreases postischemic myocardial tumor necrosis factor alpha production. Potential ultimate effector mechanism of preconditioning. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is an autocrine contributor to myocardial dysfunction and cardiomyocyte death in ischemia-reperfusion (I/R) injury, sepsis, chronic heart failure, and cardiac allograft rejection. Cardiac resident macrophages, infiltrating leukocytes, and cardiomyocytes themselves produce TNF-alpha. Although adenosine reduces macrophage TNF-alpha production and protects myocardium against I/R, it remains unknown whether ischemic preconditioning, which is mediated by adenosine, decreases postischemic myocardial TNF-alpha production. METHODS AND RESULTS: Isolated rat hearts were crystalloid perfused with the Langendorff method and subjected to global, normothermic I/R (20/40 minutes), with or without prior transient ischemic preconditioning (5 minutes) or adenosine pretreatment. Postischemic cardiac TNF alpha (ELISA) and function were determined (Langendorff). I/R increased cardiac TNF-alpha and impaired myocardial function. Ischemic preconditioning or adenosine decreased myocardial TNF-alpha and improved postischemic functional recovery. Sequestration of myocardial TNF-alpha (TNF binding protein) during the I/R experiments similarly improved postischemic myocardial function. CONCLUSIONS: This study constitutes the initial demonstration that in addition to its other beneficial effects, preconditioning decreases postischemic myocardial TNF-alpha, an autocrine contributor to postischemic myocardial dysfunction. Reduced myocardial TNF-alpha production may represent the distal effector mechanism of preconditioning. PMID- 9852906 TI - Hibernation triggers and myocardial protection. AB - BACKGROUND: Hypothermic cardioplegia provides myocellular protection, yet postischemic dysfunction remains a significant problem. Interestingly, the subcellular changes in hibernation parallel the altered biology of induced cardiac ischemia but are well tolerated by hibernated mammalian myocardium. An uncharacterized factor derived from hibernating animals, hibernation induction trigger (HIT), has been shown to induce hibernation in active animals and afford myocardial protection after ischemia-reperfusion injury. Therefore, it was of interest to further characterize the cardioprotective effects of HIT in the setting of ischemia-reperfusion injury. METHODS AND RESULTS: To determine whether HIT could improve myocardial recovery after global ischemia, isolated rabbit hearts received either standard cardioplegia or HIT in the cardioplegia or underwent preperfusion with HIT before cardioplegia. Alternatively, to determine whether HIT requires metabolic alteration, additional rabbits had in vivo pretreatment with HIT from 15 minutes to 5 days before ischemia. All hearts underwent 2 hours of global ischemia at 34 degrees C. Recovery of postischemic isovolumic developed pressure, coronary flows, and MVO2 were compared. Compared with vehicle pretreatment, HIT pretreatment (1 hour) significantly enhanced indexes of functional recovery, including developed pressure (38 +/- 3 versus 69 +/- 7 mm Hg) and coronary flow (46 +/- 2 versus 82 +/- 11 mL/min). In addition, ultrastructural morphology was preserved but only with in vivo pretreatment. Liver protein content was not increased in rabbits treated from 12 hours to 5 days with HIT versus controls, belying a protein neosynthesis mechanism. However, the temporal sequences suggested conversion of an inactive HIT profactor to an active form. CONCLUSIONS: Administration of serum derived from hibernating black bears to rabbits affords protection against ischemia-reperfusion injury compared with vehicle (saline)-treated animals in a rabbit isolated heart preparation. It is apparent that HIT deserves further identification and mechanistic study in the setting of ischemia-reperfusion injury. PMID- 9852907 TI - Phase 2 studies of adenosine cardioplegia. AB - BACKGROUND: Laboratory evidence supports the use of adenosine-supplemented cardioplegia. An initial phase 1 dose-ranging clinical evaluation demonstrated that an adenosine concentration of 15 mumol/L could be safely administered with warm blood cardioplegia and suggested that phase 2 studies were warranted. METHODS AND RESULTS: Two separate double-blind, randomized, placebo-controlled trials were performed in patients undergoing primary, isolated, nonemergent coronary artery bypass graft surgery. Patients were randomized to receive adenosine 15 mumol/L versus placebo in the first study (n = 200) and adenosine 50 or 100 mumol/L versus placebo in the second study (n = 128). Adenosine was infused with both initial and final doses of warm antegrade blood cardioplegia. The data from the 2 trials were combined using the methods of Mantel and Haenszel, and the results of the meta-analysis are presented as the relative risk with their associated 95% confidence intervals (CI). The different study groups were comparable with respect to all preoperative clinical characteristics, angiographic findings, and intraoperative variables. In both trials 1 and 2, no differences were found between groups in the incidence of the individual primary or secondary outcomes. Similarly, when both studies were combined, there was no significant evidence of any consistent treatment benefit (primary: death: relative risk [RR] = 1.02, 95% CI = 0.06, 16.6; myocardial infarction by CK-MB: RR = 0.84, CI = 0.54, 1.31; low output syndrome: RR = 1.38, CI = 0.29, 6.42; any of the above: RR = 0.98, CI = 0.78, 1.25; secondary: Q-wave myocardial infarction: RR = 1.30, CI = 0.41, 4.13; myocardial infarction by troponin T: RR = 0.7, CI = 0.40, 1.21; inotrope requirement: RR = 0.9, CI = 0.46, 1.79; intra aortic balloon pump requirement: RR = 0.6, CI = 0.07, 4.81; P > 0.20). CONCLUSIONS: Despite promising experimental data, adenosine supplementation of warm blood cardioplegia did not demonstrate any statistically significant benefit in patients undergoing elective coronary artery bypass graft surgery. Although sample sizes were relatively small, based on our interim analyses, it is unlikely that increased patient enrollment would reveal any substantive clinical differences between groups. PMID- 9852908 TI - Glucose transporter upregulation improves ischemic tolerance in hypertrophied failing heart. AB - BACKGROUND: Achieving successful myocardial preservation of hypertrophied hearts remains a difficult problem. Despite reportedly higher glycolytic potential, we have achieved limited benefit in hypertrophied hearts with strategies that successfully promote anaerobic glycolysis and long-term ischemic preservation in nonhypertrophied models. We therefore tested the hypotheses that (l) glucose transport into myocytes is a critical limiting factor in hypertrophied heart and (2) stimulation of glucose transport with vanadyl sulfate would improve postischemic recovery. METHODS AND RESULTS: Left ventricular hypertrophy in rabbits was created by aortic banding of 7- to 10-day-old rabbits. At 4 weeks of age, 1 group of animals received oral vanadyl sulfate for 3 to 4 weeks. Glucose transport (measured by the conversion of 2-deoxyglucose to 2-deoxyglucose-6 phosphate; 31P-nuclear magnetic resonance), myocardial glucose transporter content (GLUT-1 and GLUT-4 by immunoblotting), and functional recovery from ischemia-reperfusion (isolated perfused Langendorff model) were measured. Myocardial glucose transport rate was significantly reduced in hypertrophied hearts without significant reductions in glucose transporter content; these hearts were significantly less tolerant of ischemia-reperfusion than age-matched controls. Vanadyl sulfate normalized glucose transport rate and improved tolerance to ischemia-reperfusion so that postischemic function equaled that seen in controls. Lactate production during ischemia, an indication of anaerobic glycolysis, was significantly higher in hearts from vanadate-treated animals. CONCLUSIONS: Despite reportedly higher glycolytic enzyme activities, maximal glucose transport appears to be reduced and is rate limiting in hypertrophied heart. Stimulation of membrane glucose transport with vanadyl sulfate significantly improved glycolytic flux and ischemic preservation in hypertrophied hearts. PMID- 9852909 TI - Myocardial VEGF expression after cardiopulmonary bypass and cardioplegia. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a heparin-binding glycoprotein that plays a critical role in angiogenesis, vascular remodeling, and regulation of vascular tone and permeability. Because myocardial and peripheral edema and systemic hypotension occur frequently after cardiac operations, we examined the effects of cardiopulmonary bypass (CPB) and cardioplegia on gene expressions of VEGF protein and the VEGF tyrosine kinase receptor flk-1 and coronary vascular responses to VEGF. METHODS AND RESULTS: Pigs (n = 6) were placed on normothermic CPB and hearts were arrested for 1 hour with a hyperkalemic, cold blood cardioplegic solution. Pigs were then separated from CPB and perfused off CPB for an additional 2 hours. Myocardial and skeletal muscle specimens were obtained for Northern analysis of VEGF protein, flk-1 receptor, and basic fibroblast growth factor (bFGF) mRNA before CPB and after 2 hours of reperfusion. Isolated, precontracted coronary arterioles in pre-CPB dilated potently to exogenous VEGF (dilation = 26 +/- 4% of precontraction at 10(-12) mol/L VEGF). Cardioplegia-reperfusion was associated with a 4 +/- 2-fold (P < 0.05 vs pre-CPB) increase in myocardial VEGF protein mRNA, whereas no similar increase was observed in the skeletal muscle. Flk-1 mRNA was increased 6 +/- 3 fold (P < 0.05 vs pre-CPB) after reperfusion, whereas it was unchanged in the skeletal muscle. Relaxations of precontracted coronary arterioles to VEGF were significantly increased (40 +/- 6% at 10(-12) mol/L, P < 0.05 vs pre-CPB) after 2 hours of reperfusion, but those to the endothelium-dependent vasodilator ADP and the endothelium-independent vasodilator nitroprusside were not changed, suggesting that the VEGF receptors remain intact and function is selectively upregulated. In contrast, relaxation responses of microvessels to bFGF were not altered after cardioplegia-reperfusion, and there was no increase in bFGF mRNA in either myocardium or skeletal muscle. CONCLUSIONS: This study shows that VEGF protein and its flk-1 receptor gene expressions are selectively increased and the potent VEGF-induced vascular responses are enhanced in the coronary microcirculation after blood cardioplegia. The respective parameters are unchanged in the skeletal muscle after normothermic CPB. These findings may have important implications regarding postoperative coronary blood flow regulation, increases in myocardial edema, and vascular remodeling after cardioplegia reperfusion. PMID- 9852910 TI - Myocardial recovery after ischemia and reperfusion injury is significantly impaired in hearts with transgenic overexpression of beta-adrenergic receptor kinase. AB - BACKGROUND: beta-Adrenergic receptor kinase 1 (beta ARK1) mediates beta adrenergic receptor signaling via receptor phosphorylation, which results in functional uncoupling. The physiological importance of beta ARK1 on cardiac performance in the setting of ischemia and reperfusion injury, however, has not been clearly established. In this study, the effects of beta ARK1 overexpression on myocardial recovery after ischemia and reperfusion injury were evaluated in transgenic mice with the use of an isolated work-performing murine heart preparation and computerized analysis of functional data. METHODS AND RESULTS: A controlled, experimental study was performed to compare cardiac function in the hearts of both transgenic mice with a 3-fold overexpression of beta ARK1 (n = 6; weight, 25 to 29 g) and littermate controls (n = 9; weight, 25 to 28 g). Preload dependent cardiac output, contractility, heart rate, stroke work, and stroke volume were evaluated in the 2 groups before and after a 6-minute period of normothermic ischemia. Before ischemia, significant decreases were observed in all parameters of myocardial performance in beta ARK1 mice compared with control mice. After ischemia and reperfusion, significant decreases in cardiac function were observed in both experimental groups; however, significantly lower percentages of myocardial recovery occurred in beta ARK1 hearts compared with control hearts. CONCLUSIONS: After global normothermic ischemia, significant decreases in cardiac function were observed in both beta ARK1 and control mice; however, significantly lower percentages of myocardial recovery occurred in beta ARK1 mice. These data suggest that myocardial beta ARK1 overexpression significantly impairs cardiac function in the setting of ischemia and reperfusion injury. PMID- 9852911 TI - Recombinant adenovirus-mediated cardiac gene transfer of superoxide dismutase and catalase attenuates postischemic contractile dysfunction. AB - BACKGROUND: Coronary revascularization entails obligatory myocardial ischemia followed by reperfusion with occasional resultant postischemic contractile dysfunction, a state associated with significant morbidity and mortality. This injury is attributed in part to oxygen free radicals and has been partially ameliorated with exogenous antioxidants, a strategy limited by agent instability, low titer, and inadequate cardiomyocyte uptake. Cardiac gene transfer with antioxidant encoding vectors may significantly enhance intracellular free radical scavenger activity. METHODS AND RESULTS: C57/BL6 neonatal mice (age, 2 days; n = 131) underwent intrapericardial delivery of recombinant adenoviruses encoding superoxide dismutase (SOD) and catalase (Cat) (n = 76) or beta-galactosidase (LacZ) as a control (n = 55). After 3 days, hearts were explanted, and SOD and Cat transgene expression was detected by Western blot analysis. Spectrophotometric enzyme assays demonstrated enhanced SOD activity 1.6-fold (P < 0.0001) and Cat 3.6-fold (P < 0.00001) in experimental versus LacZ hearts. Isolated perfused hearts were subjected to 5 minutes of warm ischemia, and at 5, 10, and 15 minutes after initiation of reperfusion, LacZ controls lost 24%, 33%, and 41% of peak systolic apicobasal force, respectively, whereas experimental hearts lost 5%, 12%, and 20% (P < 0.001, each time point). In controls, rate of force generation diminished 8%, 17%, and 35%; in experimental hearts, it increased 1% at 5 minutes and decreased 5% and 15% and 10 and 15 minutes (P < 0.01, P < 0.05, P < 0.05). LacZ hearts exhibited dysfunction similar to hearts from uninjected animals (P = NS, each time point). CONCLUSIONS: Adenovirus mediated cardiac gene transfer and expression of SOD and Cat augment antioxidant enzyme activity and minimize contractile dysfunction after ischemic reperfusion in the isolated perfused neonatal mouse heart. PMID- 9852912 TI - In vivo transfer of a beta 2-adrenergic receptor gene into the pressure overloaded rat heart enhances cardiac response to beta-adrenergic agonist. AB - BACKGROUND: In the failing heart, the density and affinity of beta-adrenergic receptors in the myocardium both tend to decrease as the severity of cardiac dysfunction increases. If this suppression of beta-adrenergic receptors could be prevented or reversed, this could serve as a fundamental form of treatment for the failing heart. We report here a possible new therapeutic approach for the failing heart involving the transfer of the beta 2-adrenergic receptor (B2AR) gene. METHODS AND RESULTS: The B2AR cDNA was transfected in vivo to the concentric hypertrophied rat heart by intracoronary infusion with a hemagglutinating virus of Japan liposome method, and the transfected heart was transplanted into the abdomen of another rat. Four days after transfection, the sarcolemma of the cardiomyocytes was well labeled by immunohistochemical labeling. An overexpression of BAR of approximately 5 times in the heart (140 +/- 35 versus 24 +/- 3 fmol/mg protein for the transfected versus control hearts) was demonstrated by a ligand binding assay. The cardiac response of the transfected heart to isoproterenol was shown to be enhanced in a Langendorff perfusion system: After isoproterenol, developed pressure and maximal derivative of the left ventricle were greater than in the control heart (260 +/- 20 versus 230 +/- 10 mm Hg and 5500 +/- 300 versus 4500 +/- 300 mm Hg/s), and the minimal derivative of the left ventricle was remarkably smaller (-4500 +/- 300 versus 3300 +/- 200 mm Hg/s). CONCLUSIONS: These results indicate that in vivo transfection of the gene for B2AR enhances the cardiac response to isoproterenol in the pressure-overloaded rat heart (in which the disease causes a decrease in receptor density), suggesting that transfer of this gene by intracoronary infusion during cardiac arrest has potential as a novel therapeutic approach for failing hearts. PMID- 9852913 TI - Hybrid artificial lung with interleukin-10 and endothelial constitutive nitric oxide synthase gene-transfected endothelial cells attenuates inflammatory reactions induced by cardiopulmonary bypass. AB - BACKGROUND: Recently, the research on hybrid artificial organs such as heart, liver, and kidney has been reported to yield new possibilities for clinical use. However, there have been few reports on the practical use of a hybrid artificial lung, primarily due to difficulties in the concept for its application and techniques for cell attachment. In this study, we investigated the possibility for developing a novel strategy: a hybrid artificial lung with constitutive nitric oxide synthase (NOS) and interleukin (IL)-10 gene-transfected endothelial cells to attenuate inflammatory reactions induced by cardiopulmonary bypass. METHODS AND RESULTS: First, we performed an in vitro study to confirm the efficacy of our gene transfection into endothelial cells. Constitutive nitric oxide synthase and IL-10 cDNA were transfected into endothelial cells according to the hemagglutinating virus of Japan-liposome method. Levels of nitric oxide released from the endothelial constitutive NOS (ecNOS)-transfected endothelial cells were significantly higher than those of control cells (24 hours after the stimulation by lipopolysaccharide: 284.5 +/- 54.0 versus 95.7 +/- 27.9 mumol/L, P = 0.0001). On the other hand, IL-8 levels in the transfected endothelial cells were significantly lower than those in the control group (48 hours after stimulation by tumor necrosis factor-alpha: 3.1 +/- 2.4 versus 62.1 +/- 1.3 ng/mL, P = 0.0002), and IL-10 was detected in the transfected group but not in the control group. Next, we performed an in vivo study to evaluate the possibility of developing a hybrid artificial lung. One-hour partial cardiopulmonary bypass with this lung was established in rats undergoing femorofemoral bypass. Artificial lungs with no cells (group C; n = 5), that were coated with untreated endothelial cells (group E; n = 5), or that were coated with ecNOS and IL-10 gene-transfected endothelial cells (group T; n = 3) were used. At 45 minutes after the initiation of cardiopulmonary bypass, group T showed higher nitric oxide levels than groups C and E (T versus C versus E: 75.2 +/- 6.8 versus 67.2 +/- 4.3 versus 68.6 +/- 5.2 mumol/L, P = NS). The serum IL-8 levels just after cardiopulmonary bypass in group T were significantly lower than those in group C (1728 +/- 282 versus 4275 +/- 145 pg/mL, P = 0.0151). The Pao2 levels in group T just after weaning from cardiopulmonary bypass were significantly higher than those in group C (271.3 +/- 41.7 versus 136.6 +/- 12.3 mm Hg, P = 0.0362). CONCLUSIONS: These results demonstrate that a hybrid artificial lung with IL-10 and ecNOS gene-transfected endothelial cells inhibited IL-8 release and increased nitric oxide production. This suggests the possibility of developing a hybrid artificial lung capable of preserving native lung function by attenuating cardiopulmonary bypass-induced inflammatory reactions via inhibition of IL-8 release and enhanced nitric oxide production. PMID- 9852914 TI - Acute myocardial beta-adrenergic receptor dysfunction after cardiopulmonary bypass in patients with cardiac valve disease. Duke Heart Center Perioperative Desensitization Group. AB - BACKGROUND: Patients with cardiac valve disease (CVD) frequently have congestive heart failure (CHF) and chronic myocardial beta-adrenergic receptor (beta AR) desensitization. Cardiac surgery requiring cardiopulmonary bypass (CPB) is associated with increased plasma catecholamine concentrations, which might worsen myocardial beta AR function. We therefore tested the hypothesis that acute beta AR dysfunction occurs during CPB in patients with CVD. METHODS AND RESULTS: After informed consent, 50 patients were enrolled. Right atrial biopsy samples were obtained at initiation and conclusion of CPB to assess beta AR density and adenylyl cyclase (AC) activity. Plasma catecholamine concentrations increased 3 fold during CPB (P < 0.01). Although beta AR density remained constant, isoproterenol-stimulated AC activity decreased significantly (approximately 30%; P < 0.005). AC activity decreased 22% and 24% with direct G protein (NaF) or AC (manganese) activation, respectively. Patients with or without preoperative CHF exhibited similar degrees of acute myocardial beta AR dysfunction during CPB. CONCLUSIONS: Acute myocardial beta AR dysfunction occurs during CPB in patients with severe CVD requiring surgical correction, with or without preexisting CHF. The primary underlying mechanism involves functional uncoupling of the beta AR signal transduction pathway at the level of the AC moiety. This information should facilitate development of agents designed to prevent acute myocardial beta AR dysfunction during CPB, potentially leading to improved outcome in this high risk population. PMID- 9852915 TI - Inhibition of nuclear factor-kappa B nuclear localization reduces human E selectin expression and the systemic inflammatory response. AB - BACKGROUND: One proinflammatory property observed during endothelial cell activation is the expression of the neutrophil adhesion molecule E-selectin on the surface of endothelial cells. An important regulatory element in endothelial cell E-selectin expression is the nuclear localization of the transcription factor nuclear factor (NK)-kappa B, which binds to and affects the function of several genes encoding proteins mediating inflammation. METHODS AND RESULTS: In this study, we investigated the ability of pyrrolidine dithiocarbamate (PDTC), an agent that inhibits the nuclear localization of NF-kappa B, to (1) block endothelial cell E-selectin expression in vitro in response to tumor necrosis factor (TNF)-alpha, interleukin (IL)-1, and lipopolysaccharide (LPS) and (2) reduce neutrophil infiltration in a rabbit model of systemic inflammation. As measured with the use of an enzyme-linked immunosorbent assay, TNF-alpha, IL-1, and LPS each induced a significant increase in surface expression of E-selectin in cultured human umbilical vein endothelial cells (HUVECs) compared with HUVECs treated with medium alone. In contrast, E-selectin surface expression was blocked in HUVECs pretreated with PDTC before TNF-alpha, IL-1, or LPS stimulation. NF kappa B was present in HUVEC nuclei treated with TNF-alpha, whereas translocation of NF-kappa B to the nucleus was absent in TNF-alpha-treated HUVECs pretreated with PDTC. In vivo, rabbits pretreated with PDTC before LPS infusion showed significantly less neutrophil infiltration in the lungs, liver, and heart compared with animals infused with LPS alone. This correlated with a reduction in E-selectin expression in vivo. CONCLUSIONS: Our data suggest that NF-kappa B regulation of gene expression in the vascular endothelium may be an important cellular mechanism in endothelial cell activation. PMID- 9852916 TI - Evidence for unconscious memory processing during elective cardiac surgery. AB - BACKGROUND: Many anesthetic drugs have been shown to disrupt conscious recall (explicit memory) in volunteers. However, unconscious processing (implicit memory) of intraoperative auditory material may occur during general anesthesia and may provide an opportunity for intraoperative therapeutic intervention. In this study, we examined patients undergoing elective cardiac surgery for evidence of intraoperative implicit and explicit memory. METHODS AND RESULTS: Twenty-five subjects provided written informed consent and underwent general anesthesia and cardiopulmonary bypass for cardiac surgery. During the operation, patients were randomized to receive 1 of 2 different audiotapes of associated word pairs. Postoperatively, a blinded observer conducted a standardized interview to determine the extent of intraoperative implicit and explicit memory. With the use of free association, significant intraoperative implicit memory was found. In contrast, no patient had spontaneous or directed recall of intraoperative events, and we did not find evidence of intraoperative explicit memory with a recognition task. CONCLUSIONS: Patients undergoing general anesthesia for cardiac surgery were reliably able to reinforce associations between word pairs solely on the basis of their intraoperative presentation. This provides further evidence that patients are capable of processing intraoperative auditory information. PMID- 9852917 TI - Perioperative risk factors for mortality in patients with acute type A aortic dissection. AB - BACKGROUND: Patients with acute type A aortic dissection are associated with a high mortality rate and postoperative complications. This study was designed to explore perioperative risk factors for death in patients with acute type A aortic dissection. METHODS AND RESULTS: One hundred nine consecutive surgical patients with acute type A aortic dissection were included in the present study. Thirty five perioperative risk factors were used in the statistical analysis for mortality prediction. The 30-day mortality rate for all patients was 30% (33 of 109). Univariate analysis revealed 5 preoperative and intraoperative risk factors that were found to be statistically significant predictors for death: older age, renal failure, preoperative hemodynamic instability, preoperative cardiopulmonary resuscitation, and lack of retrograde cerebral perfusion (P < 0.05). Rupture of aneurysm, hypothermic circulatory arrest time, and operation date were not found to be risk factors for death. Stepwise multiple logistic regression confirmed that older age and lack of retrograde cerebral perfusion were statistically significant independent risk factors for death (P < 0.05). Four postoperative complications were found to be statistically significant prognostic indicators for death: liver failure, stroke, sepsis, and reoperation after the initial surgery (P < 0.05). CONCLUSIONS: Perioperative risk factors for death after the operation for acute type A aortic dissection were identified. This will allow physicians and surgeons to better assess the patient's risk, which will lead to better outcome. PMID- 9852918 TI - Autopsies in acute type A aortic dissection. Surgical implications. AB - BACKGROUND: This study was undertaken to review autopsy findings in operative and nonoperative settings to define anatomic pathology, cause of death, and theoretical implications of operative and perfusion management strategies for acute Stanford type A dissection. METHODS AND RESULTS: Fifty autopsies for acute type A dissection performed between 1977 and 1995 were reviewed. Twenty-nine patients had no operative therapy (group A). Twenty-one patients received an operation (group B). Cause of death in the nonoperated group was tamponade or rupture in 23 patients (23 of 29, 79%). In the operated group, 1 patient died of rupture (1 of 21, 5%). The 2 most common causes of death in the operated group were major brain injury (7 of 21, 33%) and cardiac failure (7 of 21, 33%). Intimal disruption was present in 92% of patients (45 of 49). Twenty-four percent of these (12 of 49) had a secondary intimal injury site. Theoretic femoral arterial perfusion and application of an ascending aortic cross clamp would have left 42% of patients (21 of 50) at risk of false lumen perfusion against the aortic cross clamp. Replacement of the ascending aorta alone would have left residual arch intimal disruption in 32% of patients (16 of 50). Open arch inspection and repair when appropriate would have left a minimum number of patients with distal intimal disruption (8 of 50, 16%). CONCLUSIONS: Operative therapy successfully manages the primary cause of death in acute type A dissection. A theoretical repair that uses the open arch technique followed by antegrade perfusion eliminates or minimizes the risk of false lumen perfusion in the greatest number of patients. This technique may diminish neurological injury and distal anastomosis bleeding, thereby improving outcomes. PMID- 9852919 TI - Outcomes of stent-graft treatment of false lumen in aortic dissection. AB - BACKGROUND: Stent-graft treatment for aortic disease promises to lead to a less invasive therapy than conventional surgical therapy. However, this treatment has not been established as a therapy for aortic dissection. The aim of this study was to examine the effect of stent-graft treatment of aortic dissection, as measured by follow-up data on entry site closure, clotting in the false lumen, and the outcome of the false lumen from the acute to chronic phase after operation. METHODS AND RESULTS: We used a stent-graft of our own design to close the entry site of aortic dissection in 21 cases: 9 were acute (< 14 days), 8 subacute (< 6 months), and 4 chronic (> 6 months). Nine were type A and 12 were type B. In 15 cases, the stent-graft was inserted through the transverse arch, which had been surgically opened under median sternotomy. In the other 6 cases, the stent-graft was inserted by means of a transcatheter through a femoral artery. Using computed tomographic scans, we followed and examined clot formation in the false lumen and reduction in size of the false lumen at 3 levels: at the level of maximum aortic diameter, at the distal end of the stent-graft, and 50 mm distal to the stent-graft. The entry sites were successfully closed in 19 cases (90.4%); in the remaining 2 (both treated with a transcatheter) there was perigraft leakage into the false lumen. The hospital mortality rate of these stent-graft treatments was 14.3% (3 of 21). At 2 weeks after operation, the false lumen had completely clotted, respectively, in 100%, 77%, and 38% of cases at the 3 measurement levels. Substantial shrinkage (> 50% in diameter) of the false lumen at 6 months after the operation was observed in 72%, 60%, and 38% of cases at the respective levels. Shrinkage of the false lumen was particularly enhanced in patients treated within 6 months from the onset of dissection: The false lumen shrank by > 50% in 93%, 75%, and 50% of patients. CONCLUSIONS: In cases of aortic dissection, the stent-graft is an effective tool for closing the entry site and promoting clot formation in the false lumen and for reducing the size of the false lumen within 6 months of the onset of dissection if the entry site has been closed. PMID- 9852920 TI - Retrograde cerebral perfusion results in flow distribution abnormalities and neuronal damage. A magnetic resonance imaging and histopathological study in pigs. AB - BACKGROUND: In the past few years, although significant efforts have been made to assess flow distribution during retrograde cerebral perfusion with microspheres, dye, or hydrogen clearance, flow distribution in real time is still undefined. We used MR perfusion imaging to monitor flow distribution in the brain during and after deep hypothermic circulatory arrest (DHCA) with antegrade or retrograde cerebral perfusion (ACP or RCP). METHODS AND RESULTS: Thirteen pigs were divided into 2 groups and exposed to 120 minutes of either RCP (n = 7) or ACP (n = 6) at 15 degrees C, followed by 60 minutes of cardiopulmonary bypass (CPB) at 37 degrees C. During DHCA, the brain was perfused antegradely through the common carotid artery or retrogradely through the superior vena cava at pressures of 60 to 70 mm Hg and 20 to 25 mm Hg in the ACP and RCP groups, respectively. Esophageal temperature was monitored continuously. MR perfusion images were acquired every 30 minutes before, during, and after DHCA. The brain was perfusion fixed with formaldehyde solution for histopathology at the completion of each experiment. During initial normothermic CPB, MR perfusion imaging showed a nearly uniform distribution of flow in the brain. The same pattern was maintained with a significant increase in regional cerebral blood volume during ACP and reperfusion in the ACP group. RCP provided little or no detectable blood distribution to the brain, resulting in poor reperfusion of many areas of the brain on reflow with CPB at 37 degrees C. The total area suffering poor reperfusion was significantly higher in the RCP group than the ACP group. Histopathology showed no morphological changes in any area of the brain in the ACP group, whereas varying severity of neuronal damage was observed in different regions of the brain in the RCP group. CONCLUSIONS: ACP preserves uniform blood distribution and normal morphology of brain tissue after prolonged DHCA. RCP provides very little blood to the tissue of the brain. A 120-minute period of RCP results in abnormal flow distribution and neuronal damage during reperfusion. The damage resulting from shorter periods of RCP remains to be assessed. PMID- 9852921 TI - Improvement of patency rate in heparin-coated small synthetic vascular grafts. AB - BACKGROUND: Graft thrombosis and intimal hyperplasia represent the major causes of graft failure. Heparin has been shown to have a beneficial effect on long-term patency and on prevention of intimal hyperplasia. Thus, the purpose of the present study was to evaluate the effect of heparin coating on patency rate and intimal hyperplasia in small synthetic vascular grafts. METHODS AND RESULTS: Two synthetic grafts (expanded polytetrafluoroethylene [ePTFE], and polyurethane) with and without heparin coating were implanted in the infrarenal aorta (diameter, 2 mm) of 40 Whistar rats. Animals survived 8 weeks after implantation. Graft patency, intimal thickness, and percentage of diameter stenosis were determined by light microscopy at the proximal respectively distal anastomosis and in the middle of the graft. Uncoated grafts showed a patency rate of 70% for ePTFE and 60% for polyurethane grafts. Heparin-coated grafts showed a patency rate of 100% for ePTFE and 90% for polyurethane grafts. Intimal hyperplasia was observed in all grafts mainly at the anastomosis site. Intimal wall thickness and percentage of stenosis were significantly more pronounced in the polyurethane than ePTFE grafts (P < 0.01). Heparin coating significantly reduced overall graft thrombosis (P < 0.05) but had no significant effect on intimal hyperplasia. CONCLUSIONS: Small grafts show a high rate of graft thrombosis and an enhanced intimal hyperplasia. ePTFE grafts show significantly less intimal hyperplasia and percentage of stenosis than polyurethane grafts. Heparin coating significantly reduced graft thrombosis but had no significant effect on intimal hyperplasia. Thus, heparin coating seems to be beneficial for graft patency, and ePTFE appears to be superior to polyurethane as graft material. PMID- 9852922 TI - Topography of cell replication in human vein graft stenoses. AB - BACKGROUND: Analysis of the cellular composition of human autogenous vein graft lesions at the time of revision provides an opportunity to identify the cellular processes leading to the development of stenosis in humans after vascular reconstruction. METHODS AND RESULTS: Human vein graft-threatening stenotic lesions were identified by duplex scanning within 3 to 18 months after infrainguinal bypass and surgically removed. They were serially studied by immunocytochemistry for expression of the proliferating cell nuclear antigen (PCNA) in different cell types: alpha-actin-positive smooth muscle cells (SMCs), endothelial cells (ECs), monocytes, and macrophages. Proliferation indexes were separately obtained for each layer of the vessel wall by determining the mean percentage of PCNA-positive nuclei among the total number of nuclei present within the intima, the media, and the adventitia, respectively. The percentage distribution of the replicating cell types was also determined. We report that in autogenous vein graft (n = 14) the intima of the lesion displayed fewer PCNA + nuclei (1.03 +/- 0.88) than the underlying media (3.14 +/- 0.74) or the adventitia (3.01 +/- 0.74). Replicating SMCs were predominantly in the medial layer (68% of PCNA + cells) of stenotic vein grafts. In the adventitia, the proliferation was most intense in the endothelium of microvessels (65% of PCNA + nuclei). CONCLUSIONS: Our findings reveal a 3-fold greater proliferative activity in the media and the adventitia as compared with the intima of autogenous vein graft lesions, in contrast to cellular proliferation identified in recurrent coronary stenotic plaques. Moreover, there are distinctive patterns of distribution of the different cell populations among the 3 layers. The results indicate a proliferative response of the media and the adventitia of autogenous vein grafts transplanted into the arterial circulation, in addition to the cellular proliferation observed in the intima of the lesion. PMID- 9852923 TI - Immunohistochemistry of matrix metalloproteinases and their inhibitors in thoracic aortic aneurysms and aortic valves of patients with Marfan's syndrome. AB - BACKGROUND: Thoracic aortic aneurysms (TAAs) and valvular insufficiency, the main cardiovascular lesions in Marfan's syndrome, are associated with destruction of connective tissue; however, their pathogenesis remains unclear. METHODS AND RESULTS: To test the hypothesis that changes in the activity of the matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are responsible for the damage to connective tissue in these lesions, histochemical studies of the immunoreactivity (IR) for MMPs and their tissue TIMPs (MMP-1, MMP-2, MMP-3, MMP 9, TIMP-1, and TIMP-2) were made in TAAs (n = 7) and aortic valves (n = 5) from 7 patients with Marfan's syndrome. All TAAs showed cystic medial necrosis (CMN), with loss of elastic fibers and smooth muscle cells. Extensive areas of myxoid change were found in all aortic valves. Areas of CMN showed no IR for any MMPs or TIMPs. The IR of smooth muscle cells at the borders of areas of CMN was stronger for all MMPs, especially MMP-2 and MMP-9, than in other regions. The surfaces of disrupted elastic fibers showed IR for MMP-2 and MMP-9. Areas of myxoid change showed similar but less pronounced alterations. CONCLUSIONS: We hypothesize that the defect in fibrillin-1 in Marfan's syndrome leads to (1) formation of elastin that is abnormally aggregated and more easily degraded by MMPs than is normal elastin, (2) upregulation of the synthesis of MMPs, (3) progressive destruction of connective tissue by these enzymes, and (4) development of TAAs and valvular lesions. PMID- 9852924 TI - Nonthrombogenic, adhesive cellular lining for left ventricular assist devices. AB - BACKGROUND: The textured, blood-contacting surfaces of the Thermocardiosystems HeartMate left ventricular assist device (LVAD) promote the passivation of the biomaterial caused by the accumulation of an integral coagulum. Commonly, acute, postimplantation thrombocytopenia causes significant bleeding, requiring surgery or blood transfusions. Chronic complications include thromboembolic microevents that can affect central nervous system function. Pumps, explanted during donor organ transplantation, are often found to have an extensive cellular panus associated with the blood-contacting surfaces of the device. This natural cellular lining suggests a possible strategy for improving the blood biocompatibility of the HeartMate. Therefore, seeding of LVADs with cells genetically engineered to enhance their antithrombotic properties before implantation was investigated as a means to improve biocompatibility for long term use. METHODS AND RESULTS: Bovine vascular smooth muscle cells genetically engineered to produce nitric oxide were seeded on LVAD biomaterials and exposed to elevated shear stresses to determine cell-adhesive capabilities. Comparative studies were performed with vascular endothelial cells isolated from the same vessel. To assess the thrombogenic potential of the genetically engineered smooth muscle cells, monolayers were exposed to whole blood in parallel plate flow chambers and were platelet-adhesion quantified. This procedure used scanning electron microscopy and computer image-capture software. Endothelial cell monolayers and mock-transduced smooth muscle cells were assayed in a comparative manner. LVADs were seeded with genetically engineered smooth muscle cells and maintained under cell culture conditions for 96 hours. Thereafter, seeded LVADs were incorporated into in vitro flow loops. Cell retention within the pump was determined by sampling the effluent culture medium downstream of the pump and cell counting in a Coulter counter. After 18 hours of in vitro flow, a seeded pump was implanted into the abdominal cavity of a calf and anastomosed to the apex of the heart and to the descending aorta. More genetically engineered smooth muscle cells were retained on the surface of LVAD biomaterials when they were subjected to shear stresses up to 75 dyne/cm than endothelial cells assayed in the identical manner. Adherence of platelets to the surface of smooth muscle cells was significantly reduced after their transduction with nitric oxide synthase with GTP cyclohydrolase genes. Platelet deposition on the genetically modified myocyte layers was similar to that associated with endothelial cell layers. Cell loss from cell-seeded LVADs incorporated into in vitro flow loops remained < 5% of the total cell number seeded regardless of the duration of flow. CONCLUSIONS: LVADs seeded with smooth muscle cells, transduced with the genes to optimize nitric oxide production, adhered well to the pump surface under in vitro and in vivo flow conditions. PMID- 9852925 TI - Long-term dynamic cardiomyoplasty improves chronic and acute myocardial energetics in a model of left ventricular dysfunction. AB - BACKGROUND: We present the first long-term evaluation of myocardial energetics after dynamic cardiomyoplasty (CMP) in a model of left ventricular (LV) dysfunction. METHODS AND RESULTS: Seventeen dogs underwent rapid ventricular pacing (RVP) to create heart failure. Eight dogs were randomly selected to undergo cardiomyoplasty. All dogs continued RVP for 6 additional weeks, whereas the CMP dogs underwent a simultaneously delivered synchronized muscle wrap conditioning protocol. After termination of RVP at 10 weeks in all dogs, myoplasty dogs continued to receive muscle wrap stimulation until the terminal study. Pressure-volume analysis to assess LV energetics was conducted at baseline and 4 weeks and 3 months after termination of RVP (6 months after baseline). At 6 months, CMP dogs displayed enhanced contractility, lower volumes, and more optimal energetics compared with control animals. Acute muscle wrap stimulation further increased effective contractility and myocardial efficiency compared with unassisted beats. CONCLUSIONS: The decrease in NYHA functional class that occurs in patients after dynamic cardiomyoplasty may be secondary to its beneficial effects on long-term myocardial function, volume, and energetics. PMID- 9852926 TI - Sinus node function after a systematically staged Fontan procedure. AB - BACKGROUND: Sinus node dysfunction has been previously reported to occur in 13% to 16% of patients after the Fontan operation. Although there is concern that an intermediate cavopulmonary connection may increase the risk of sinus node dysfunction, previous studies have not reported on patients routinely staged to a Fontan operation. This study sought to determine the early and late incidences of sinus node dysfunction in patients systematically and uniformly staged to a Fontan operation after a prior hemi-Fontan. METHODS AND RESULTS: To determine the early incidence of sinus node dysfunction, hospital records and perioperative ECGs were reviewed in all 287 patients having had a staged Fontan operation between January 1990 and December 1995. A cross-sectional analysis was performed on 220 of 239 surviving patients (92%) to determine the late incidence of sinus node dysfunction. Sinus node dysfunction was present in 7% of the patients before and in 15% after the hemi-Fontan. Although most patients (81%) regained normal sinus node function between the 2 stages, 23% had sinus node dysfunction in the early postoperative period after the Fontan. Of the 95 patients followed for > 4 years after the Fontan operation, 44% had sinus node dysfunction. However, at a mean follow-up of 3.5 +/- 1.7 years, only 16 patients (6.7%) had received a pacemaker and only 10 (4.1%) had documented atrial flutter. CONCLUSIONS: Perioperative sinus node dysfunction is common after both the hemi-Fontan and the Fontan procedures. Although many patients regain sinus node function between the 2 stages, late sinus node dysfunction is common and more likely to occur in patients with early sinus node dysfunction and those with longer follow-up. PMID- 9852927 TI - Biventricular repair for double-outlet right ventricle. Results and long-term follow-up. AB - BACKGROUND: The purpose of the present study was to define the optimal management and to identify the risk factors for death and repeat operation in patients with double-outlet right ventricle. METHODS AND RESULTS: From 1985 through 1996, 154 consecutive patients underwent biventricular repair for double-outlet right ventricle. The presence of bilateral infundibular structures was the major inclusion criteria (142 patients). According to the relationship of the ventricular septal defect (VSD) to the great arteries, there were 86 patients with a subaortic VSD (56%), 45 patients with a subpulmonary VSD (29%), 18 patients with a noncommitted VSD (12%), and 5 patients with a doubly committed VSD (3%). Sixty-five patients (42%) had undergone previous palliative procedures. At repair, the median age was 10 months, and the median weight was 6.5 kg. Two main types of repair were used: intraventricular baffle repair (n = 115) and arterial switch operation with VSD-to-pulmonary artery baffle (n = 39). There were 14 hospital deaths (9%; 70% confidence limit [CL], 7% to 12%). The only significant risk factor for early death was the presence of congenital mitral valve anomalies (P = 0.02). Twenty-eight patients (18%) required 39 repeat operations. The repeat operation rate was higher in patients with associated VSD enlargement at baffle construction (n = 29; 19%) (P = 0.01). There were 6 late deaths (4%; 70% CL, 2% to 7%). Patients presenting with pulmonary stenosis constituted a low-risk group for global death (P = 0.008). The median follow-up was 52 months. Ten-year actuarial survival and survival with freedom from repeat operation rates were 86% and 62% (70% CL, 83% to 89% and 54% to 70%), respectively. CONCLUSIONS: Long-term survival with good quality of life can be achieved after either 1- or 2-stage repair of this complex anomaly. PMID- 9852928 TI - New approach for reconstruction of the pulmonary outflow tract during the Ross procedure. AB - BACKGROUND: This study describes and evaluates the early results of a new surgical technique to reconstruct the right ventricular outflow tract and fit it with a new valve after the Ross procedure. METHODS AND RESULTS: Between March 1996 and November 1997, 20 patients underwent a Ross operation with a new approach consisting of a direct anastomosis between the remaining pulmonary artery trunk and the infundibulum and of the creation of a monocusp tailored from the anterior pulmonary artery wall as a means of reconstructing the valve. The 20 patients (12 males and 8 females) had a mean age of 27.4 years (range, 17 to 42 years). Ten of them had predominant aortic regurgitation, 8 had aortic stenosis, and 2 had mixed disease. There were no deaths during the follow-up period of up to 20 months, no early or late repeat operations, and no specific complications secondary to the surgical technique. During the follow-up, at the aortic autograft site, 19 patients had no or trivial regurgitation, and a mild regurgitation was found in only 1 patient. Across the pulmonary monocusp, color flow Doppler demonstrated no or trivial incompetence in 10 patients, mild incompetence in 7, and moderate incompetence in 3. No significant pressure gradient was shown. CONCLUSIONS: Our experience supports the use of this new surgical procedure and allows extension of the Ross operation to where there are no facilities for homografts. It may be an alternative for right ventricular outflow tract reconstruction with a homograft should the results be confirmed at long-term follow-up. PMID- 9852929 TI - Hyperventilation impairs oxygenation after bidirectional superior cavopulmonary connection. AB - BACKGROUND: Bidirectional superior cavopulmonary connection (BSCC) may be complicated by systemic hypoxemia. Hyperventilation, which is standard therapy for postoperative hypoxemia, has opposing effects on the pulmonary and cerebral vascular beds, which are connected after BSCC. It is unknown which of these effects predominates and, therefore, whether hyperventilation improves or impairs systemic oxygenation after BSCC. METHODS AND RESULTS: Twelve consecutive patients (median age, 6.4 months; age range, 6.0 to 32.0, months) undergoing BSCC were studied prospectively. Patients were studied in the intensive care unit within 6 hours of surgery and while sedated, paralyzed, and mechanically ventilated. Inotropes were not altered, and no transfusions were given. FIO2 was set at 100%, and peak end-expiratory pressure was set at 0. Each patient was studied first during normal ventilation, then during hyperventilation, and finally again during normal ventilation. Hyperventilation resulted in significant decreases in arterial PO2, systemic oxygen saturation, and transpulmonary gradient. Cerebral blood flow velocity was measured in 6 patients through transcranial Doppler sonography of the middle cerebral artery. Mean cerebral flow velocity decreased significantly during hyperventilation. CONCLUSIONS: Hyperventilation significantly impairs systemic oxygenation after BSCC. This fall in oxygenation occurs despite a decrease in transpulmonary gradient. A possible mechanism for this effect is that hyperventilation lowers arterial PCO2, raising cerebral vascular resistance, and lowering cerebral, superior vena caval, and pulmonary blood flows. Supportive evidence for this mechanism is the decrease in cerebral flow velocity that occurs during hyperventilation. After BSCC, normal ventilation rather than hyperventilation should be used to improve systemic oxygen levels. PMID- 9852930 TI - High-volume continuous hemofiltration during cardiopulmonary bypass attenuates pulmonary dysfunction in neonatal lambs after deep hypothermic circulatory arrest. AB - BACKGROUND: Cardiopulmonary bypass (CPB) induces an inflammatory reaction that activates neutrophils and releases free radicals in tissue. Ischemia-reperfusion further aggravates inflammation. Hemofiltration (HF) could potentially remove inflammatory mediators and reduce injury. This study assessed the effect of continuous high-volume HF during CPB on systemic edema formation and pulmonary function after deep hypothermic circulatory arrest (DHCA). METHODS AND RESULTS: Anesthetized lambs (n = 16) underwent CPB with systemic cooling (40 minutes), DHCA (120 minutes at 18 degrees C), and rewarming (40 minutes). All animals were weaned from CPB and observed for 3 hours after reperfusion. Continuous HF was used in 8 lambs at a flow rate of 300 mL/kg per hour throughout CPB, simultaneously replacing hemofiltration volume with a balanced salt solution (HF group). In 8 control animals, no hemofiltration was performed. Hematocrit remained at 23% to 25% during the experiment in both groups. Pulmonary vascular resistance (PVR), lung dynamic compliance (Cdyn), alveolar-arterial oxygen difference (AaDO2), and total body water content (bioimpedance) were measured. Malondialdehyde (MDA), a product of lipid peroxidation, was assayed in lung tissue. Percent increase of body water content at 180 minutes of reperfusion was significantly lower in the HF group than in control (132 +/- 2% vs 152 +/- 5%, P < 0.005). There was less of a rise in PVR compared with baseline at 180 minutes of reperfusion in the HF group than in control (131 +/- 8% vs 238 +/- 26%, P < 0.005). In addition, percent recovery of Cdyn and AaDO2 in the HF group was preserved significantly (respectively, P < 0.05) 2 hours after reperfusion than in the control group. Lung tissue MDA in the HF group (46.2 +/- 12.6 vs 65.3 +/- 17.1 nmol/L per gram of tissue, P < 0.05) was significantly lower than in the control group. CONCLUSIONS: High-volume, continuous hemofiltration during CPB attenuates systemic edema formation, pulmonary hypertension, the extent of lung dysfunction, and depression of cardiac output and reduces free radical-mediated tissue injury after CPB with DHCA. This technique may have a clinical application to reduce the morbidity rate of CPB. PMID- 9852931 TI - Neurological function after deep hypothermic circulatory arrest in the rat. AB - BACKGROUND: Integrated neurological function, behavior, and somatic recovery were studied in 35 rats undergoing 5 to 80 minutes of hypothermic circulatory arrest (HCA). METHODS AND RESULTS: A closed extracorporeal circulation system (ECC) consisting of a miniature oxygenator and heat exchanger, primed with 6 mL of asanguinous solution, was connected to a closed-chest rat with cannulae in the right atrium for venous drainage (ID = 1.7 mm) and in the ascending aorta for arterial return (ID = 1.0 mm). The rat was surface- and core-cooled until rectal temperature reached 18 degrees C, when ECC was stopped and cardioplegic solution delivered. After 5, 10, 20, 40 (each n = 5), and 80 minutes (n = 15) of HCA, the rat was reperfused, weaned from ECC, and followed with behavioral scoring, passive avoidance tasks, and cardiopulmonary exercise testing until euthanized for morphological study. Every rat resumed weight gain in the first week after HCA and regained preoperative exercise capacity by the fourth week. Only rats undergoing 80 minutes of HCA showed behavioral abnormalities such as stereotypy and incomplete righting reflex, which eventually disappeared in the fourth week. Learning ability was preserved in all except for rats after 80 min of HCA, who failed to acquire new memory to avoid electric stimuli (n = 10) up to 3 months after HCA, when pyramidal cells were partly replaced by astroglia in the cerebral cortex and CA1 sector of hippocampus. Nonetheless, old memory established before HCA was preserved even after 80 minutes of HCA and allowed rats (n = 5) to avoid electric stimuli. CONCLUSIONS: Homogeneity of animals, miniature ECC system, and an established testing system allowed evaluation of rats after HCA, which disclosed learning disability (functional disorder) and pyramidal cell loss (organic defect) after 80 minutes of HCA despite recovery of somatic function, behavior, and growth. PMID- 9852932 TI - Effects of a monoclonal antibody to P-selectin on recovery of neonatal lamb hearts after cold cardioplegic ischemia. AB - BACKGROUND: The interaction between endothelium and leukocytes plays a crucial role in ischemia-reperfusion injury. P-selectin, which is expressed on activated endothelium, mediates the first step in leukocyte adherence to the endothelium. This study examined the effects of a monoclonal antibody (mAb) against P-selectin on the recovery of cardiac function and myocardial neutrophil infiltration after ischemia. METHODS AND RESULTS: Thirteen blood-perfused, isolated neonatal lamb hearts underwent 2 hours of hypothermic cardioplegic arrest and 2 hours of reperfusion. Immediately before reperfusion, mAb to P-selectin was administered to the perfusate (15 micrograms/mL) in 6 hearts (group P-sel). In control (n = 7), the same volume of saline was added. Isovolumic left ventricular function and coronary blood flow were measured. At 2 hours after reperfusion, myocardial myeloperoxidase activity, an index of neutrophil accumulation, was assayed. At 30 minutes of reperfusion, hearts treated with mAb to P-selectin achieved significantly greater recovery of maximum developed pressure (70 +/- 4% in control versus 77 +/- 2% in group P-sel, P < 0.01), maximum positive first derivative of pressure (dP/dt) (64 +/- 7% in control versus 73 +/- 5% in group P sel, P < 0.05), and maximum negative dP/dt (61 +/- 6% in control versus 70 +/- 6% in group P-sel, P < 0.05) compared with control. Percent baseline of coronary blood flow was also significantly increased in group P-sel (135 +/- 40% in control versus 205 +/- 43% in group P-sel, P < 0.05). Myocardial myeloperoxidase activity was significantly lower (P < 0.05) in group P-sel (4.7 +/- 3.2) versus control (16.0 +/- 10.1). (Units are change in absorbance/min/g tissue.) CONCLUSIONS: The functional blockade of P-selectin resulted in better recovery of cardiac function and attenuated neutrophil accumulation during early reperfusion. Strategies to block P-selectin mediated neutrophil adherence may have clinical application in improving myocardial function at early reperfusion. PMID- 9852934 TI - Canada to welcome new guide to physical activity. PMID- 9852933 TI - Maze procedure for atrial fibrillation associated with atrial septal defect. AB - BACKGROUND: Atrial fibrillation (AF) is a relatively common complication in elderly patients with an atrial septal defect (ASD). However, use of the simultaneous maze procedure for AF associated with ASD remains controversial. We examined the efficacy and risk of the use of the maze procedure in these patients. METHODS AND RESULTS: Between March 1992 and April 1997, 26 patients underwent the maze procedure as a concomitant operation with ASD closure (maze group). Kosakai's modified maze procedure was performed in 17 patients, the modified Cox maze II or III procedure was performed in 6, and the restrictive right-sided maze procedure was performed in 3. The mean +/- SD age at surgery was 58.2 +/- 9.1 years. The mean +/- SD duration of AF was 7.8 +/- 8.5 years. The mean +/- SD left atrial dimension was 47 +/- 9 mm, and 24 patients (92%) had a larger-than-normal (> 35 mm) left atrium. The mean +/- SD follow-up period was 2.7 +/- 1.7 years. There were no hospital or late deaths. There was no thromboembolic episode in the late follow-up period. Sinus rhythm was regained in all patients who underwent conventional the right- and left-sided maze procedure except for 1 patient. However, 3 patients who underwent the restrictive right sided maze procedure showed a return to AF rhythm. The atrial A wave was detected with the use of pulsed Doppler study in all patients who had a restored sinus rhythm. During the same period, 45 patients who were > 40 years old and without AF underwent only ASD closure (control group). The incidence of reopening the chest for bleeding was significantly (P = 0.046) higher in the maze group (12%) than in the control group (0%). Paroxysmal AF more frequently (P = 0.023) occurred in the control group (18%) than in the maze group (0%). CONCLUSIONS: These results suggest that the standard maze procedure should be considered in patients with AF associated with ASD. The restrictive right-sided maze procedure was not reliable, probably due to preoperative enlargement of the left atrium. PMID- 9852935 TI - Outcomes of coronary artery bypass surgery in elderly people. AB - OBJECTIVES: To document the impact of coronary artery bypass (CABG) surgery on quality of life in elderly people three months after surgery; to compare quality of life measures; to examine predictors of poor quality of life; and to assess the predictive validity of global clinical rating scales. STUDY DESIGN: Prospective cohort study with postoperative, three-month and one-year follow-up. SETTING: A 700-bed tertiary care teaching hospital in Halifax, Nova Scotia. PATIENTS: One hundred of 200 consecutive patients, 75 years and older, undergoing CABG. MEASURES: Demographic information, clinical data and global clinical ratings were collected at baseline to assess risk for adverse outcome (death, stroke, functional impairment) at each follow-up. Quality of life three months post-CABG was documented using the RAND 36-Item Health Survey and the Seattle Angina Questionnaire. MAIN RESULTS: Perioperative deaths occurred in four patients, disabling strokes in six and postoperative complications in seven. Three additional deaths occurred by the three-month follow-up and nine more by one year. On average, important improvements in quality of life were observed at three months' follow-up, using both assessments of quality of life. CONCLUSIONS: Many elderly patients do well after CABG surgery, but a higher proportion than is seen in younger patients have complications leading to death and disability. A global clinical measure by the attending cardiologist at baseline did not identify patients at a higher risk. Follow-up studies are needed to assess long term outcomes of older patients at higher risk of death or a poor quality of life after CABG surgery. PMID- 9852936 TI - The effect of early ambulation on hematoma formation and vascular complications following 7 French diagnostic cardiac catheterization. AB - OBJECTIVE: To assess the feasibility and safety of early ambulation 3 to 4 h after diagnostic 7 French cardiac catheterization. DESIGN: Randomized, single blind assignment to one of 3, 4 or 6 h ambulation postcardiac catheterization groups. SETTING: Tertiary care community hospital in an urban region. PATIENTS: Eight hundred and seventy-four consecutive inpatients and out-patients presenting for routine diagnostic cardiac catheterization. INTERVENTION: Hematoma formation and other vascular complications recorded at the time of discharge and 24 h later. MAIN RESULTS: No significant difference in hematoma formation rates was noted among patients mobilized at 3 h (3.6%), 4 h (4.8%) or 6 h (3.2%). Late hematoma formation occurred in 2.3% of patients. Other vascular complications were very rare. Reported rates of hematoma formation varied significantly (P < 0.05) among physicians, ranging from 0.9% to 8.0%. CONCLUSIONS: Early ambulation of patients 3 to 4 h after routine diagnostic 7 French cardiac catheterization is both safe and feasible. These findings could result in more efficient recovery bed utilization, reduced nursing costs and improved patient compliance with bed rest. PMID- 9852937 TI - Prehospital management of acute myocardial infarction: Electrocardiogram acquisition and interpretation, and thrombolysis by prehospital care providers. AB - OBJECTIVE: To review prehospital management of patients with suspected ST elevation acute myocardial infarction (AMI) based on the acquisition and interpretation of electrocardiograms (ECGs), and the effects of thrombolytic therapy initiated by prehospital care providers. DESIGN: MEDLINE was searched by combining the search phrases 'thrombolysis,' 'paramedics' and 'myocardial infarction' to identify all pertinent articles. The bibliographies were reviewed to search for other relevant articles. RESULTS: The earlier that treatment is initiated in AMI, the better the prognosis. Multiple randomized and nonrandomized trials indicate that prehospital care providers (including paramedics, nurses and doctors) are able to acquire prehospital ECGs with negligible increases in on scene time, ranging from 30 s to 7 mins. With minimal training, they are capable of accurately interpreting ECGs and diagnosing ST elevation AMI, with results comparable with control ECGs obtained by physicians. Numerous studies have investigated the role of specially trained prehospital personnel in initiating thrombolysis. Trials outside of North America have predominantly used physicians, whereas North American studies employed paramedics. Thrombolysis has been shown to be safe and effective when started outside the hospital by physicians or paramedics, with a reduction in time to treatment and no increase in complications. The further a patient with ST elevation AMI is from hospital, the greater the potential benefit of prehospital thrombolysis. The European Myocardial Infarction Project (EMIP), the largest randomized trial of prehospital thrombolysis, demonstrated a trend towards reduced mortality but was underpowered to detect significant mortality differences. The Grampian Region Early Anistreplase Trial (GREAT), a rural study, is the only randomized trial to demonstrate a statistically significant mortality difference in patients receiving prehospital thrombolysis. Despite trends in favour of prehospital diagnosis and treatment of AMI, no urban study has been sufficiently powered to demonstrate mortality benefits. CONCLUSION: Prehospital treatment of patients with chest pain using ECGs and thrombolysis is safe. Though rural patients have significant reductions in total mortality when treated with thrombolysis in a prehospital setting, this has not been documented with an urban population. Prehospital identification of thrombolysis-eligible patients with ST elevation AMI via acquisition and interpretation of ECGs followed by triage to a hospital 'lytic team' has the potential to improve patient outcome and requires further investigation. A prehospital paramedic program for identifying and treating thrombolysis-eligible patients requires intensive planning, retrospective feasibility work, implementation and monitoring to establish effectiveness. PMID- 9852938 TI - Atrial fibrillation: overview of therapeutic trials. AB - Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia treated by physicians, and a plethora of therapeutic trials deal with selected aspects of its management. This overview attempts to categorize and summarize the available studies. A key to management of AF is a clinical classification schema that provides a framework for application of the available treatment modalities. Such a classification is provided. Antithrombotic trials have demonstrated the remarkable efficacy of warfarin and more modest effect of acetylsalicylic acid for prevention of stroke; these data are summarized. Cardioversion to restore sinus rhythm is an important aspect of management of AF, particularly of persistent and new onset AF. In this review pharmacological cardioversion is emphasized. The data concerning the use of various drugs for pharmacological cardioversion are reviewed. Many, but not all, agents have been shown to have efficacy in this regard, but efficacy with drugs is lower than that with electrical cardioversion and, in the case of amiodarone, may be delayed. For recurrent AF, the two major rhythm management approaches are maintenance of rhythm and heart rate control. Trials of pharmacological and nonpharmacological therapies for these purposes are reviewed and summarized. Management of AF is an active area of research, and the present review is intended as a foundation upon which new information can be added. PMID- 9852939 TI - Technology and application of ultraminiature catheter pressure transducers. AB - After a brief historical account of the methods for pressure measurements in the cardiovascular system, the basic structural elements of a new generation of miniaturized catheter pressure transducers are described. These catheters have an outside diameter at the tip of 0.9 mm (3 French) and have been routinely applied in left and right heart catheterization in intact, anesthetized rats. Together with cardiac output measured by the thermodilution technique, a complete set of basal functional parameters can be obtained in vivo. The method of cardiac catheterization in rats is accurate, reliable and easy to perform. As to left heart function, changes occurring in several models of cardiac hypertrophy and heart failure have been recorded and correlated with morphological and metabolic alterations. In addition, the functional effects of catecholamines and thyroid hormones have been evaluated. In addition to the routine catheterization procedure, a double catheter method has been introduced recently, which allows measurement of left ventricular isovolumetric pressure in intact rats. Catheterization of the right ventricle requires a more refined catheter with a characteristic bend at the tip so that it can be comfortably slid from the right atrium into the right ventricle. With this method it was found that right ventricular systolic pressure was elevated markedly in rats with chronic myocardial infarction induced by ligation of the left anterior descending coronary artery, by pulmonary artery banding, by intermittent chronic hypoxia and by noradrenaline administration. The ultraminiature catheter pressure transducer has also been successfully applied in an isolated working rat heart preparation. Recent modifications of this kind of catheters also enabled the catheterization of the left ventricle in mice. Future applications of ultraminiature catheter pressure transducers may be directed to catheterization of the pulmonary artery in rats and to the in vivo and in vitro assessment of heart function of transgenic mice. PMID- 9852940 TI - Atrial fibrillation: what are the effects of drug therapy on the effectiveness and complications of electrical cardioversion? AB - Atrial fibrillation is the most common cardiac rhythm disorder associated with hospitalization. Two therapeutic options have been available: antiarrhythmic drug therapy, and external or internal electrical cardioversion. Electrical cardioversion of atrial fibrillation remains one of the most widely used and effective treatments for the restoration of normal sinus rhythm. However, many patients continue to receive an antiarrhythmic drug before and after cardioversion in an attempt either to cardiovert the arrhythmia chemically or to maintain sinus rhythm after successful cardioversion. Because some pharmacological agents can affect the cardioversion procedure for atrial fibrillation or flutter, and because many patients with such arrhythmias may require electrical cardioversion when they are taking antiarrhythmic drugs, the question of a possible effect of drug therapy on the efficacy and safety of electrical cardioversion of atrial fibrillation arises. Early reports of direct current cardioversion provoking potentially lethal ventricular arrhythmias raised suspicions of an arrhythmogenic role for digoxin antiarrhythmic therapy, and it is customary to withhold these drugs for 24 to 48 h before cardioversion is attempted. However, this complication is likely to arise only in patients who are close to, or actually manifesting, signs of drug toxicity. On the other hand, treatment with therapeutic concentrations of antiarrhythmic drugs before cardioversion may in some cases be associated with a significant reduction in the number of shocks and decreased energy required to restore sinus rhythm, a lower incidence of postshock arrhythmias and a reduced risk of early recurrence of atrial fibrillation. PMID- 9852941 TI - Possible adverse skin reaction to carvedilol. AB - As experience with beta-blocker use in patients with congestive heart failure is increasing, enthusiasm for carvedilol use in these patients is also increasing. Dermatological reactions to carvedilol are rare and have not been well described in the literature. A patient who developed an acute dermatological reaction after one dose of 3.125 mg carvedilol is presented. Given the rapid onset of symptoms experienced by the patient after a single, small dose of carvedilol, clinicians should be aware of the potential adverse effects of the drug. PMID- 9852942 TI - Primary cardiac angiosarcoma with superior vena caval syndrome: review of surgical resection and interventional management of venous inflow obstruction. AB - Primary cardiac angiosarcoma occurs rarely, and surgical resection is often required to relieve symptoms. A 66-year-old man whose presenting symptoms were palpitations and general fatigue is presented. Echocardiography revealed a large tumour occupying most of the right atrium. When superior vena caval (SVC) syndrome developed, surgical resection of the tumour was attempted. The tumour was found to have invaded the inflow of SVC, left atrium, right ventricle and ascending aorta. The bulk of the tumour was resected, requiring reconstruction of the right atrium and caval inflows. Histopathological diagnosis of the tumour was primary angiosarcoma. In the postoperative period, symptoms of SVC syndrome recurred, which were relieved temporarily by balloon angioplasty. After one month, recurrence of symptoms was again managed by catheter dilation. On this occasion a metallic stent was deployed, which prevented further symptoms of SVC syndrome during the eight months before the patient died with generalized metastases. PMID- 9852943 TI - The Drosophila ebony gene is closely related to microbial peptide synthetases and shows specific cuticle and nervous system expression. AB - The previously detected ebony (e) locus (Caizzi et al., 1987) consists of a complex gene structure that is divided into seven exons. An open reading frame encoding the putative Ebony protein of 98.5 kDa exhibits homology to a family of peptide synthetases (Stachelhaus and Marahiel, 1995), in good correlation with the proposed function as beta-alanyl-dopamine synthetase. Multiple ebony transcripts are detected throughout development. P-factor mediated transformation of genomic DNA rescues the cuticle, electrophysiological and behavioural phenotypes. Fusion of the ebony reading frame with that of beta-galactosidase of E. coli reveals expression in cuticle and nervous system. Strong staining in the first and, to a lesser extent, in the second optic neuropile may reflect the pronounced visual defect observed in ebony mutants. In addition, weak central brain and thoracic ganglion expression is detected in flies. Conservation of a multidomain protein structure known from peptide synthetases should have functional implications on the putative reaction mechanism of peptide bond formation. PMID- 9852944 TI - Identification of the gene and the protein of RNA polymerase II subunit 9 (Rpb9) from the fission yeast Schizosacharomyces pombe. AB - Both the rpb9 gene and its cDNA encoding the subunit 9 of RNA polymerase II were cloned from the fission yeast Schizosaccharomyces pombe. From the DNA sequences, Rpb9 was predicted to consist of 113 amino acid residues with a molecular mass of 13,175. S. pombe Rpb9 is 47, 40 and 36% identical in amino acid sequence to the corresponding subunits from Saccharomyces cerevisiae, human and Drosophila melanogaster, respectively. Previously, we failed to detect Rpb9 in the purified RNA polymerase II by amino-terminal micro-sequencing of proteolytic fragments of subunits separated by SDS-gel electrophoresis. After Western blot analysis using antibodies raised against the protein product of the newly isolated rpb9 gene, we found that the purified RNA polymerase II contains Rpb9. PMID- 9852945 TI - Isolation of a nitrogen response regulator gene (nrr1) from Metarhizium anisopliae. AB - Attempts to improve the effectiveness of entomopathogenic fungi as biological control agents require a clear understanding of the pathogenicity determinants at both the biochemical and molecular level. Proteases play a key role in entomopathogenicity, allowing the fungus to penetrate the insect cuticle and rapidly invade the host. The most extensively studied of these protease activities, PR1A and PR2, are both subject to nitrogen derepression. The Metarhizium anisopliae nrr1 (nitrogen response regulator 1) gene was identified using a PCR-based strategy; it encodes a putative DNA-binding protein with a single zinc finger motif defined by the C-X2-C-X17-C-X2-C sequence. M. anisopliae NRR1 shows a significant sequence similarity to Neurospora crassa NIT2. Sequence analysis identified the presence of two introns, suggesting a greater degree of similarity to N. crassa nit2 than to the areA-like genes that have been identified. However, functional equivalence of nrr1 to areA was demonstrated, by co-transformation and complementation of an A. nidulans areA loss-of-function mutant (areA18 argB2 pabaA1 inoB2) with the M. anisopliae nrr1 gene. The areA /nrr1+ Aspergillus transformants were able to grow on media with nitrate and glutamate as the sole nitrogen source, whereas the areA- strain is unable to grow under these conditions. The possible relevance of nitrogen regulation to pathogenicity is discussed. PMID- 9852946 TI - A new membrane-bound OprI lipoprotein expression vector. High production of heterologous fusion proteins in gram (-) bacteria and the implications for oral vaccination. AB - We have previously described the development of cloning vectors for the production of OprI-based outer membrane fusion proteins in E. coli (Cornelis et al., 1996) and now describe the construction of a new vector, containing a lacI(q) gene, resulting in tight repression of the promotor and allowing its use in other Gram (-) bacteria. The new pVUB3 expression vector encodes a truncated but active LacI(q)(341) repressor which binds to the single operator in the vector. A high repression of the trc promotor was observed, resulting in a very low basal leakage of expression and very high production levels of OprI or derivatives after IPTG induction in E. coli. Bacterial viability was not affected under uninduced conditions, but the number of viable cell counts decreased after production of large amounts of the outer membrane-bound OprI lipoprotein and its derivatives, both in E. coli and Salmonella typhimurium. This highly repressible system allows us to extend the use of OprI vectors in other Gram (-) bacteria, resulting in the production of outer membrane-bound lipid-modified molecules, opening the possibility for its application in the design of potential live Salmonella-based subunit vaccines. PMID- 9852947 TI - Cloning vectors for the expression of green fluorescent protein fusion proteins in transgenic plants. AB - A series of versatile cloning vectors has been constructed that facilitate the expression of protein fusions to the Aequorea victoria green fluorescent protein (GFP) in plant cells. Amino-terminal- and carboxy-terminal protein fusions have been created and visualized by epifluorescence microscopy, both in transgenic Arabidopsis thaliana and after transient expression in onion epidermal cells. Using tandem dimers and other protein fusions to GFP, we found that the previously described localization of wild-type GFP to the cell nucleus is most likely due to diffusion of GFP across the nuclear envelope rather than to a cryptic nuclear localization signal. A fluorescence-based, quantitative assay for nuclear localization signals is described. In addition, we have employed the previously characterized mutants GFP-S65T and GFP-Y66H in order to allow for the expression of red-shifted and blue fluorescent proteins, respectively, which are suitable for double-labeling studies. Expression of GFP-fusions was controlled by a cauliflower mosaic virus 35S promoter. Using the Arabidopsis COP1 protein as a model, we confirmed a close similarity in the subcellular localization of native COP1 and the GFP-tagged COP1 protein. We demonstrated that COP1 was localized to discrete subnuclear particles and further confirmed that fusion to GFP did not compromise the activity of the wild-type COP1 protein. PMID- 9852948 TI - The exon/intron organization of the globin gene of Scapharca inaequivalvis homodimeric hemoglobin: unusual intron homology with other bivalve mollusc globin genes. AB - In this study, we have investigated the positions of introns in the globin gene of Scapharca inaequivalvis homodimeric hemoglobin. We found the three exon/two intron organization typical of vertebrate globin genes, with the two introns in highly conserved positions, as it occurs in the A and B globin genes of the tetrameric hemoglobin from the same organism, confirming the absence of the so called 'central intron' found in the globin genes of plants and of some invertebrates. We identified two homodimeric globin genes (3207 and 2723 bp) that differ only with respect to the size of the first intron. Sequence analysis of the two first introns (1668 and 1364 bp) has revealed that they are highly homologous, except for a 569- and 296-bp insertion in each intron I. Interestingly, the two first introns contain regions with an unusually high identity (approximately 80%) with regions of the first intron of the congeneric clam Anadara trapezia and the related clam Barbatia reveana globin genes, suggesting that these uncoding regions may have played a regulatory role that has subsequently been lost during the course of the evolution. PMID- 9852949 TI - Molecular cloning of bovine telomerase RNA. AB - Telomerase is a ribonucleoprotein reverse transcriptase essential for the maintenance of telomere length. However, the available information concerning the biochemical and structural aspects of mammalian telomerases is scarce, primarily due to the low abundance of these enzymes and the difficulty and expense involved in its purification. To overcome these problems, we started to purify and characterize telomerase from bovine testis. Bovine telomerase was purified over columns of hydroxyapatite, DEAE-Sepharose, heparin-agarose, phenyl-agarose and spermine-agarose. In a sedimentation study performed using a 15-40% glycerol gradient, partially purified bovine telomerase exhibited an apparent molecular weight of more than 1000 kDa. One 435-bp RNA, bTR, was cloned from the most purified telomerase fraction and shown to be co-purified with telomerase activity in a glycerol gradient. bTR shares 83% similarity to the human telomerase RNA and 65% to the mouse telomerase RNA. A putative template region encompassing 10 nucleotides (5'-CUAACCCUAA-3') complementary to the mammalian telomere sequence (TTAGGG)n is located between nucleotides 38-47 of bTR. Besides, the bTR 5' flanking region shares only three regulatory elements with that of hTR: a TATA like sequence, a CCAAT box and an E1A-F box. Furthermore, the addition of in vitro transcribed bTR reconstituted telomerase activity after removal of the endogenous bTR by micrococcal nuclease digestion. Northern blot analysis identified a single RNA of about 430-440 nucleotides expressed in the bovine testis and an immortalized bovine cell line. Taken together, these data strongly suggest that bTR is the RNA component of bovine telomerase. PMID- 9852950 TI - Vectors for the expression of tagged proteins in Schizosaccharomyces pombe. AB - A series of vectors is described which enables the episomal expression of proteins fused to different tag sequences in Schizosaccharomyces pombe. Proteins can be expressed with their amino termini fused to GFP/EGFP, three copies of the HA or Pk epitopes or a combined tag which contains two copies of the myc epitope and six histidine residues (MH). Fusion of the carboxyl terminus of a protein to a tag is possible with GFP/EGFP or Pk. Expression of the fusion proteins is controlled by the medium strength mutant version of the regulatable nmt1 promoter. PMID- 9852951 TI - Constitutive and barbital-induced expression of the Cyp6a2 allele of a high producer strain of CYP6A2 in the genetic background of a low producer strain. AB - The levels of one or more cytochrome P450 (CYP) enzymes and the respective mRNAs are found to be higher in insecticide-resistant insects than in susceptible insects. To understand better how insects regulate the levels of CYPs, we examined the expression of the Cyp6a2 gene in various strains of Drosophila melanogaster. We also took a transgenic approach to understand the molecular mechanisms that are involved in strain variation of Cyp6a2 expression. RNA blot analysis showed that the constitutive expression of Cyp6a2 varies from strain to strain; the level of CYP6A2 mRNA is barely detectable in the underproducer ry506 strain, whereas it is very high in the overproducer 91-R and MHIII-D23 strains. The long terminal repeat (LTR) of mobile element 17.6 that is found in the 3' untranslated region (UTR) of the Cyp6a2 gene of some strains does not appear to have any role on the steady-state CYP6A2 mRNA level. We also found that the Cyp6a2 gene is inducible by barbital in 91-R, ry506 as well as 91-C, which carries an LTR insertion. To examine the genetic background of the underproducer ry506 strain with respect to Cyp6a2 expression, we transformed the ry506 strain with the Cyp6a2 allele of the overproducer 91-R strain (Cyp6a2-91 R) and measured the constitutive and barbital-induced expression of the Cyp6a2-91 R transgene in the transformed flies. The Cyp6a2-91 R transgene carrying 129 bp of DNA upstream of the ATG codon did not show any constitutive or barbital-induced expression in the ry506 host genome. However, transgenes with 1331 and 985 bp upstream DNA showed similar levels of constitutive expression that were higher than that of the endogenous Cyp6a2 gene of the ry506 host strain, but lower than the expression of the same gene in the 91-R strain. Both these transgenes, with 1331 and 985 bp upstream DNA, also showed induction with 0.1 M barbital. DNA sequence analysis revealed that in both 91-R and ry506, the upstream DNA between +1 and 985 bp contains a distal and a proximal group of three potential barbie boxes, i.e. cis-elements that are thought to be involved in barbiturate-mediated induction of CYP genes. Except for four bases located near the distal cluster of barbie boxes and two other bases, the base sequence of the upstream DNA is identical in ry506 and 91-R strains. These results suggest that the underproducer ry506 strain has the trans-regulatory factors to support constitutive and induced expression of the Cyp6a2-91 R allele carrying DNA between -129 and -1331 bp regions. Possible reasons for low constitutive expression of the endogenous Cyp6a2 gene and moderate level of expression of the Cyp6a2-91 R allele in the ry506 genetic background are discussed. PMID- 9852952 TI - Mapping protein-ligand interactions using whole genome phage display libraries. AB - The function of many genes cannot be deduced from sequence similarity, and biochemical methods are usually required. Whole genome sequences can be thought of as not only a set of genes but also collections of functional domains. These domains can be studied by affinity methods whereby identification of the ligand can provide information on biochemical function. To take advantage of this method, one must express all functional domains in a form suitable for affinity studies. Phage display technology provides a means for accomplishing this. The pJuFo phage display system, based on the interaction between the leucine zippers Jun and Fos, has been modified and used to create a genomic phage display library from Escherichia coli MG1655. The system has been tested by using the library to map the dominant binding epitopes for an anti-RecA protein polyclonal antibody sera. This methodology provides a general biochemical approach to functional analysis of protein-ligand interactions on a genomewide basis. PMID- 9852953 TI - Germ cell-specific enhancer activity of a repeated element in a variable region of the mouse genome. AB - We recently described a complex genetic structure on mouse chromosome 8, a region of the murine genome in which genetic rearrangements frequently occur. A large repeated element specific to this chromosome was found to overlap with one of the cadherin genes (Cad11). An additional degree of complexity became apparent with the identification, in a subset of laboratory strains of mice, of a retrogene integrated into one of the repeated units. Designated Sycp1-ps2, it originated from the early meiotic gene encoding Synaptonemal Complex Protein 1. We now report that, among wild Mus species in which the retrogene is not present, this region of Chr 8 shows a high degree of variability. Sequence analysis showed that integration of Sycp1-ps2 created a 5' transcription initiator element. Transcription of the pseudogene in the testis was directly demonstrated. A germ cell-specific enhancer activity was localized within a 1117 bp region of the repeat, which was sufficient to direct the expression of reporter genes in transgenic mice to late meiotic and post-meiotic spermatogenic cells. PMID- 9852954 TI - Generation of expressed sequence tags as physical landmarks in the genome of Trypanosoma brucei. AB - Previous molecular genetic studies on the African trypanosome have focused on only a few genes and gene products, the majority of which are concerned with surface antigenic variation; consequently, an insignificant number of the genes of this organism have been characterized to date. In order to: (1) identify new genes and analyze their expression profile, (2) generate expressed sequence tags (ESTs) for derivation of a physical map of the trypanosome genome, and (3) make available the partial sequence information and the corresponding clones for general biomedical research on the parasite, we have performed single-pass sequencing of random, directionally cloned cDNAs from a bloodstream form Trypanosoma brucei rhodesiense library. Analysis of 2128 such ESTs sequenced so far in this study showed significant similarities [BLASTX P(n)-value < 10(-4), and a match > 10 amino acid residues] with proteins whose genes have been described in diverse organisms including man, rodents, kinetoplastids, yeasts and plants. A number of the ESTs encode homologues of proteins involved in various functions including signal reception and transduction, cell division, gene regulation, DNA repair and replication, general metabolism, and structural integrity. Although some of these genes may have been expected to be present in the African trypanosomes, the majority of them had not previously been described in these organisms. A large proportion, 768 individual ESTs (36%, representing 385 different transcripts), had a significant homology with genes described in organisms other than the African trypanosomes; however, 15% of the ESTs were from genes already described in trypanosomes. Among the ESTs analysed were 462 distinct known genes, only 77 of which have been described in T. brucei. Approximately 52% of the ESTs did not show any significant homology with the sequences in any of the public domain databases. PMID- 9852955 TI - New alternatively spliced variants of calmodulin-dependent protein kinase II from rabbit liver. AB - Polymerase chain reaction analysis revealed four alternatively spliced variants of each of the gamma and delta isoforms of calmodulin-dependent protein kinase II (CaM-kinase II) in rabbit liver. Among the four variants of the gamma isoform, two were novel ones, designated as CaM-kinase II gamma-H and gamma-I. The gamma-I variant possessed both of the two deletable exons; D2a and D2b, which had never been found together in any variant. Sequence analysis of the gamma-I indicated that the D2a was upstream of the D2b and that they were contiguous with each other in the gamma-I. Among the four variants of the delta isoform, two were also novel ones, designated as CaM-kinase II delta-11 and delta-12, and the other two were the already-reported ones, delta-2 and delta-6. The delta-11 and delta-12 were identical to the delta-2 and delta-6, respectively, except that three bases (CAG) located at a splicing junction was deleted in the delta-11 and delta-12, suggesting two splicing sites of a single intron. Thus, the diverse splicing patterns may produce many more variants than those so far considered. PMID- 9852956 TI - Isolation of an Arabidopsis thaliana cDNA by complementation of a yeast abc1 deletion mutant deficient in complex III respiratory activity. AB - The yeast Abc1 protein acts as a chaperone-like protein essential for the proper conformation and efficient functioning of the respiratory complex III. By functional complementation of a yeast abc1 mutant, we have identified an Arabidopsis thaliana cDNA that corresponds to a single copy gene and encodes a protein sharing 45% similarity with the yeast Abc1p protein. Cytochrome spectra and respiratory activity measurements have shown that the plant protein allows a partial restoration of the complex III activity. No major difference in the steady-state level of ABC1At mRNA was observed in various plant tissues, suggesting that ABC1At is constitutively expressed in A. thaliana. Phylogenetic analysis revealed that the Abc1At protein belongs to a large family of proteins composed of two eukaryotic and one prokaryotic subgroups differing by their degree of similarity and probably by their function. PMID- 9852957 TI - Molecular cloning of Drosophila melanogaster cDNAs that encode a novel histone deacetylase dHDAC3. AB - The steady-state level of histone acetylation in eukaryotes is established and maintained by multiple histone acetyltransferases (HATs) and histone deacetylases (HDACs) and affects both the structure and the function of chromatin. Histone deacetylases play a key role in the regulation of transcription, and form a highly conserved protein family in many eukaryotic species. Here we describe the cloning, sequencing and genetic mapping of two histone deacetylase genes in Drosophila melanogaster: dHDAC1 is essentially identical to the previously cloned D. melanogaster d-Rpd3 gene and dHDAC3, a novel gene, is orthologous to the human and the chicken (Gallus gallus) HDAC3 genes. The predicted amino acid sequence (438 aa) of dHDAC3 shows 58.1% identity with dHDAC1/d-Rpd3, the only previously known member of the HDAC family in this organism. The map positions on polytene chromosomes for dHDAC1 and dHDAC3 were determined as 64C1-6 and 83A3-4 respectively. A search for other dHDAC3-like genes failed to find other potential paralogues in D. melanogaster, but identified significant homologies with bacterial and fungal genes encoding enzymes that metabolise acetyl groups, and with genes for other hydrolyases such as carboxypeptidase. In addition, histone deacetylase activity in D. melanogaster nuclear extracts can be inhibited by high concentrations of zinc and activated by low concentrations, which is identical to the properties of bovine carboxypeptidase A. On the basis of sequence and functional similarities, we suggest that histone deacetylases are metal substituted enzymes. PMID- 9852958 TI - Molecular cloning of FKHRL1P2, a member of the developmentally regulated fork head domain transcription factor family. AB - Here we report the expression of a fork head domain protein in human T helper cells. We cloned and characterized a fork head cDNA from human T helper cell mRNA using differential display RT-PCR. The cDNA contains a 546-nucleotide (nt) open reading frame (ORF) that codes for the carboxyl-terminal 180 amino acids (aa) of the recently identified fkhrl1 gene. This ORF does not contain the characteristic DNA-binding domain found in members of the forkhead protein family. In-vitro transcription/translation of this cDNA expressed a protein of approximately 20 kDa. We have generated antibodies that specifically immunoprecipitated the in vitro-translated 20-kDa protein. This antibody also recognizes in human T lymphocytes a 70-kDa protein corresponding in size to that predicted for the fkhrl1 gene product. The mRNA levels for fkhrl1 is elevated in T helper-induced lymphocytes in comparison to PHA-stimulated T lymphocytes. Further characterization of FKHRL1 and its related family members should shed light on the transcriptional mechanisms of this fork head gene subfamily and their role in T helper cell differentiation and regulation of cell growth. PMID- 9852960 TI - Cloning and characterization of Planctomyces limnophilus rpoN: complementation of a Salmonella typhimurium rpoN mutant strain. AB - The rpoN gene, which encodes the alternative sigma factor sigma 54, was cloned from the budding, peptidoglycan-less bacterium Planctomyces limnophilus. P. limnophilus rpoN complemented the Ntr- phenotype of a Salmonella typhimurium rpoN mutant strain. The P. limnophilus rpoN gene encoded a predicted polypeptide that was 495 residues in length and shared a significant homology with other members of the sigma 54 family. The protein sequence displayed all of the characteristic motifs found in members of this family, including the C-terminal helix-turn-helix motif and the well-conserved RpoN box. A potential sigma 54-dependent activator was also identified in P. limnophilus. These findings extend the range of phylogenetic groups within the Domain Bacteria that are known to contain sigma 54. PMID- 9852959 TI - Cloning, bacterial synthesis, and characterization of immunoglobulin variable regions of a monoclonal antibody specific for the hepatitis B virus X protein. AB - The nucleotide (nt) sequences encoding the variable regions of the heavy (H) and light (L) chains were determined for a murine monoclonal antibody, 12/231/93, which is specific for a linear epitope located between amino acids 90 and 102 of the hepatitis B virus (HBV) X protein (HBx). The variable (V) regions of the H and L chains were shown to belong to the mouse H chain subgroup II (C) and kappa L chain group III, respectively. The cloned variable region sequences were used for the production of a Fab fragment in Escherichia coli, which had binding activity for membrane immobilized recombinant HBx. These gene sequences may be useful for the study of HBx function in cells that will support HBV replication. PMID- 9852962 TI - CINEMA--a novel colour INteractive editor for multiple alignments. AB - CINEMA is a new editor for manipulating and generating multiple sequence alignments. The program provides both an interface to existing databases of alignments on the Internet and a tool for constructing and modifying alignments locally. It is written in Java, so executable code will run on most major desktop platforms without modification. The implementation is highly flexible, so the applet can be easily customised with additional functions; and the object classes are reusable, promoting rapid development of program extensions. Formerly, such extended functionality might have been provided via browser plug-ins, which have to be downloaded and installed on every client before loading data. Now, for the first time, an applet is available that allows interactive client-side processing of an alignment, which can then be stored or processed automatically on the server. The program is embedded in a comprehensive help file and is accessible both as a stand-alone tool on UCL's Bioinformatics Server; http:/(/)www.biochem.ucl.ac.uk/bsm/dbbrowser+ ++/CINEMA2.02/, and as an integral part of the PRINTS protein fingerprint database. Exploitation of such novel technologies revolutionises the way users may interact with public databases in the future: bioinformatics centres need not simply provide data, but are now able to offer the means by which information is visualised and manipulated, without the requirement for users to install software. PMID- 9852961 TI - Molecular cloning of XNLRR-1, a Xenopus homolog of mouse neuronal leucine-rich repeat protein expressed in the developing Xenopus nervous system. AB - We report the isolation and characterization of a Xenopus sequence, XNLRR-1, that is closely related to a gene for mouse neuronal leucine-rich repeat protein (NLRR 1). The cDNA clone is 4179 bp long and encodes a putative transmembrane glycoprotein of 718 amino acids, containing 12 leucine-rich repeats followed by one C2-type immunoglobulin-like domain and one fibronectin type-III repeat. XNLRR 1 is transcribed mainly in the developing eye area and the ventricular zone from diencephalon to hindbrain and slightly in spinal cord in Xenopus tadpoles. The similarity of the XNLRR-1 gene to other known cell adhesion molecules, together with the expression pattern, suggests that XNLRR-1 is involved in interactions at the neuronal cell surface. PMID- 9852963 TI - Can functional regions of proteins be predicted from their coding sequences? The case study of G-protein coupled receptors. AB - A filter based on a set of unsupervised neural networks trained with a winner take-all strategy discloses signals along the coding sequences of G-protein coupled receptors. By comparing with the existing experimental data it appears that these signals correlate with putative functional domains of the proteins. After protein alignment within subfamilies, signals cluster in protein regions which, according to the presently available experimental results, are described as possible functional domains of the folded proteins. The mapping procedure reveals characteristic regions in the coding sequences common and/or characteristic of the receptor subtype. This is particularly noticeable for the third cytoplasmic loop, which is likely to be involved in the molecular coupling of all the subfamilies with G-proteins. The results indicate that our mapping can highlight intrinsic representative features of the coding sequences which, in the case of G-protein coupled receptors, are characteristic of protein functional regions and suggest a possible application of the filter for predicting functional determinants in proteins starting from the coding sequence. PMID- 9852964 TI - Varicella-zoster virus glycoprotein gE: endocytosis and trafficking of the Fc receptor. AB - Varicella-zoster virus (VZV) encodes a functional cell membrane Fc receptor called glycoprotein gE. VZV gE resembles other mammalian cell membrane receptors, such as the mammalian Fc receptor. In further analyses by transient transfection, the cellular trafficking of VZV gE was compared to other cell surface receptors. VZV gE was shown to undergo endocytosis from the cell membrane when visualized by laser scanning confocal microscopy. The endocytosis and trafficking pathway of VZV gE followed closely the pathway defined for the human transferrin receptor. Receptor-mediated endocytosis of VZV gE was dependent on a YAGL motif in its cytoplasmic tail. In addition, VZV gE underwent receptor-mediated endocytosis when it bound the Fc portion of immunoglobulin. Thus, this detailed study of VZV gE cellular trafficking has revealed potential roles for gE during viral infection. PMID- 9852965 TI - Intracellular transport of varicella-zoster glycoproteins. AB - Previous observations have established that varicella-zoster virus (VZV) is enveloped in the trans-Golgi network (TGN) in cultures infected with VZV and that the glycoprotein gE is targeted to the TGN by a signal sequence (AYRV) and an acidic TGN signal patch in its cytosolic domain. Neither sequence is present in other VZV glycoproteins. Like gE, gI was targeted to the TGN when it was expressed in transfected cells, suggesting that gI also contains TGN targeting information (colocalized with gE and the AP-1 adaptin complex). In contrast, gB, gC, gH, and gL immunoreactivities were not detected in the TGN when they were expressed individually in transfected cells. In VZV-infected cells, gE, gI, gH, and gL were all concentrated in the TGN. Since VZV glycoproteins that lack targeting sequences (gB, gC, gH, and gL) concentrated in the TGN of infected cells, it is proposed that gE and gI, which have such sequences, serve as navigator glycoproteins, forming complexes that direct the signal-deficient glycoproteins to the TGN. PMID- 9852966 TI - Baculovirus expression, purification, and properties of varicella-zoster virus gE, gI, and the complex they form. AB - Varicella-zoster virus (VZV) expresses six known glycoproteins. High level expression of recombinant soluble forms of the VZV glycoproteins E and I (gE and gI) was achieved in the baculovirus system. gE and gI associate in VZV-infected cells to form an intermolecular complex. To purify large amounts of these glycoproteins, gE was produced with a C-terminal six-histidine (HIS-6) tag sequence, and gI was produced both with and without the HIS-6 sequence. The individual glycoproteins or the gE/gI complex were purified in their native forms by use of affinity chromatography. Recombinant soluble VZV gE and gI provided important tools in the biochemical analysis and may contribute further to the functional and immunologic studies of these VZV envelope components. PMID- 9852967 TI - Regulated nuclear localization of the varicella-zoster virus major regulatory protein, IE62. AB - The major transcriptional regulatory protein encoded by varicella-zoster virus (VZV), IE62, accumulates within the nucleus of transfected and VZV-infected cells. Data are presented to show that nuclear localization of IE62 is dependent upon charged amino acids mapping to residues 677-685 of the 1310 residue protein. Furthermore, coexpression of VZV open-reading frame (ORF) 66 with IE62 results in the accumulation of cytoplasmic forms of IE62, suggesting that the ORF 66 protein can override the IE62 nuclear localization signal. Consistent with this, cytoplasmic forms of IE62 were detected in VZV-infected cells at late but not immediate early times after infection. The ORF 66 protein, a putative protein kinase, did not affect nuclear localization of other proteins, and IE62 nuclear localization was not affected by coexpression with VZV ORF 47, another putative kinase. These results suggest that IE62 nuclear functions may be specifically regulated by the VZV ORF 66 protein. PMID- 9852968 TI - Analysis of the glycoproteins I and E of varicella-zoster virus (VZV) using deletional mutations of VZV cosmids. AB - The contributions of the glycoproteins gI (ORF67) and gE (ORF68) to varicella zoster virus (VZV) replication were investigated in deletion mutants made by using cosmids with VZV DNA derived from the Oka strain. These experiments demonstrated that gI was not required for VZV replication in vitro but gE appeared to be. Although VZV gI was not required, its deletion or mutation resulted in a significant decrease in infectious virus yields, and it disrupted syncytial formation and altered the conformation and distribution of gE in infected cells. Normal cell-to-cell spread and replication kinetics were restored when gI was expressed from a non-native locus in the VZV genome. The expression of intact gI, the ORF67 gene product, is required for efficient VZV replication. PMID- 9852969 TI - Anatomy of the varicella-zoster virus open-reading frame 4 promoter. AB - The regulation of varicella-zoster virus (VZV) gene expression is largely controlled at the transcriptional level by a few key viral proteins cooperating with one another and with cellular transcription factors. However, the mechanisms involved are largely unclear. To identify the sequences important for the transcriptional regulation of open-reading frame (ORF) 4, itself encoding a transcriptional regulator, a mutation analysis of the promoter was done. These studies identified an element between -69 and -59 (relative to the transcriptional start site), which was critical to the activity of the promoter upon stimulation by the VZV transactivator IE62 and by VZV infection. DNA-protein interaction studies revealed that VZV induced the binding of a specific protein complex to this element, which contained the ubiquitous transcription factor USF. ORF 4 is the second VZV gene (in addition to VZV ORF 29) in which USF binding plays a critical role in gene expression. PMID- 9852970 TI - The varicella-zoster virus (VZV) open-reading frame 29 protein acts as a modulator of a late VZV gene promoter. AB - Transient expression assays have shown that the varicella-zoster virus (VZV) open reading frame (ORF) 29 gene product can act as a modulator of VZV gene expression. The ORF 29 protein alone does not appear to have any effect on transcription; however, in its presence, changes in the level of reporter gene activity mediated by the VZV immediate early (IE) 62 major transactivator are seen. Increased expression was observed in human fibroblasts, MeWo cells, HeLa cells, and T cells. In contrast, the presence of the ORF 29 protein results in a down-regulation of IE62 activation in PC-12 rat neuronal cells. Competition filter binding assays indicate that the ORF 29 protein binds specifically to the glycoprotein I promoter. Since transcripts for ORF 29 and ORF 62 have been detected in latently infected ganglia, the gene regulatory properties of the ORF 29 protein may be relevant to maintenance or establishment of VZV latency. PMID- 9852971 TI - Isolation and utilization of human dendritic cells from peripheral blood to assay an in vitro primary immune response to varicella-zoster virus peptides. AB - A human dendritic cell-based assay used to monitor a T cell proliferation response to viral peptides in vitro is described. Dendritic cells and autologous CD4+ T cells were isolated from peripheral blood by a series of density-gradient centrifugations or magnetic bead separations (or both). Peptides corresponding to residues of the immediate early protein, IE62, of varicella-zoster virus (VZV) were used as stimulating antigens, and persons with no history of varicella and no humoral or cellular immunity to VZV served as naive donors for the assays. Three VZV-susceptible donors were tested, and all demonstrated an in vitro response to multiple VZV peptides. This assay has potential as a screen to establish the immunogenicity of viral antigens in vitro using T cells from naive donors. PMID- 9852972 TI - Varicella-zoster virus IE63, a virion component expressed during latency and acute infection, elicits humoral and cellular immunity. AB - Varicella-zoster virus (VZV) latency in human dorsal root ganglia is characterized by the transcription of large regions of its genome and by the expression of large amounts of some polypeptides, which are also expressed during lytic cycles. The immediate early 63 protein (IE63) is a virion component expressed very early in cutaneous lesions and the first viral protein detected during latency. Immune response against IE63 has been evaluated among naturally immune adults with a history of chickenpox: Specific antibodies were detected in serum, and most subjects who had a T cell proliferation with unfractionated VZV antigens had T cell recognition of purified IE63. The cytotoxic T cell (CTL) response to IE63 was equivalent to CTL recognition of IE62, the major tegument component of VZV, whose immunogenicity has been previously described. T cell recognition of IE63 and other VZV proteins is one of the likely mechanisms involved in controlling VZV reactivation from latency. PMID- 9852973 TI - Evidence of latent varicella-zoster virus in rat dorsal root ganglia. AB - Latent varicella-zoster virus (VZV) was studied in ganglia of rats that had been inoculated subcutaneously with either a high-passaged wild-type, a low-passaged wild-type, or the vaccine strain of virus using in situ hybridization. Nine of 11 rats injected with virus and no control rats developed serum VZV antibodies as demonstrated by fluorescent antibody membrane antigen. Polymerase chain reaction 2 weeks following inoculation did not detect viremia in the rats. VZV was detected by in situ hybridization in ganglia of 10 of the 11 infected rats but not in ganglia of the control rats. The distribution of VZV DNA is identical to that seen in humans; satellite cells and neurons contain VZV DNA. Although all animals received unilateral injections of virus, VZV DNA was in ipsilateral and contralateral ganglia in 6 animals, suggesting that virus replication and viremia had occurred. PMID- 9852974 TI - Adult susceptibility to varicella in the tropics is a rural phenomenon due to the lack of previous exposure. AB - In the industrialized countries of the West, varicella is largely a childhood disease, whereas reports from tropical countries indicate a significant incidence of varicella among adults. High ambient temperature, epidemiologic interference from other viruses, and race have been blamed. We tested our hypothesis that less exposure to varicella-zoster virus (VZV) during childhood in rural areas accounts for the reported greater frequency of varicella in adults in tropical climates by comparing rates of VZV seropositivity among urban and rural adult Bengalee populations living in identical climatic conditions in India. Only 5 (3.4%) of 153 urban adults were seronegative compared with 74 (31.1%) of 246 rural adults. Ninety-six percent of urban adults were immune by the age of 25, compared with 42% in the rural group. The results suggest that higher adult susceptibility to varicella is seen only in rural areas of the Tropics and is due to reduced transmission of VZV. PMID- 9852975 TI - Single amino acid change in DNA polymerase is associated with foscarnet resistance in a varicella-zoster virus strain recovered from a patient with AIDS. AB - The genetic characterization of a foscarnet-resistant strain of varicella-zoster virus (VZV) that was isolated from a patient with AIDS is reported. Compared with the sequence of the Dumas reference strain, this strain, which was isolated from a patient in whom foscarnet treatment failed, had two point mutations. The emergence of one of the mutations, which includes a change from a glutamic acid to a lysine at position 512 in the DNA polymerase, suggests that this mutation is implicated in the VZV foscarnet resistance. The other mutation, which replaces serine 863 by a glycine, is also present in 2 susceptible strains--Oka and a wild type isolate. PMID- 9852976 TI - Cost of varicella in France: a study in day care centers. AB - To assess the cost of varicella in young children in France, a prospective study was done in day care centers. Children (1263), who were 3 months to 3 years old and attending day care, were followed over a varicella season. For every child who developed varicella (n = 200), detailed information was obtained by use of parental questionnaires. Questions concerned medical care, days missed from work for parents, and the need for extra baby-sitting. On average, each sick child had one consultation with a physician and received three medications. In half of the families (52%), at least 1 parent had to miss work an average of 4.5 days to care for a sick child. Total costs to society were estimated to be US$352 per family, with medical costs accounting for 22% of the cost. The average eventual cost to parents was $89 per family, including $80 of non-medical costs. This study emphasizes the important socioeconomic impact of varicella in the day care setting in France. PMID- 9852977 TI - Polymerase chain reaction and restriction fragment length polymorphism analysis of varicella-zoster virus isolates from the United States and other parts of the world. AB - A polymerase chain reaction (PCR) assay that identifies and differentiates wild type (wt) and vaccine strains of varicella-zoster virus (VZV) was used to determine if VZV strains with restriction fragment length polymorphisms resembling those of the Japanese Oka vaccine strain were present in the wt pool outside of Japan. Virus samples (n = 114) from patients with chickenpox and zoster from various parts of the United States and Australia were analyzed. The assay correctly identified 113 samples as wt strain. The 1 sample identified as Oka vaccine strain came from a child with leukemia who developed a vaccine associated rash after receiving the live attenuated varicella vaccine. At this point, there is no evidence that wt strains resembling the vaccine are circulating outside of Japan. This indicates that this PCR assay can be utilized to distinguish rashes due to vaccine and wt VZV. PMID- 9852978 TI - Racial and psychosocial risk factors for herpes zoster in the elderly. AB - The effects of black race and psychologic stress on the risk of acquiring herpes zoster in late life were examined. Subjects were participants of a stratified probability sample of community-dwelling persons > or = 65 years old. A comprehensive health survey was administered in 1986-1987 (P1), 1989-1990 (P2), and 1992-1994 (P3). Incident cases of zoster between P1 and P2 and P2 and P3 served as the dependent variables. Hypothesis-testing variables included race, negative life events, and measures of social support. Control variables included age, sex, education, cancer, other chronic diseases, hospitalization, activities of daily living, self-rated health, depression, and cigarette smoking. From P1 to P2, 1.4% of black and 3.4% of white subjects developed zoster (P < .001). From P2 to P3, 2.9% of black and 7.5% of white subjects developed zoster (P < .001). After controlling for variables, black subjects were significantly less likely to develop zoster than were white subjects (adjusted odds ratio, 0.37; 95% confidence interval, 0.26, 0.53; P = .0001). Most measures of stress were not significantly related to zoster; however, study limitations preclude definitive conclusions. PMID- 9852979 TI - The identification of risk factors associated with persistent pain following herpes zoster. AB - Demographic and clinical characteristics of patients with herpes zoster at the time of presentation predict the duration and severity of pain on long-term follow-up. Analyses by Cox's proportional hazard models of six databases from controlled trials of antiviral drugs (total subjects = 2367) identified covariates for zoster-associated pain; all tests for significance were two-sided. Age strongly influenced pain outcome: patients > or = 50 years old were significantly more likely to have prolonged zoster-associated pain compared with those < 30 years old. Patients with prodromal symptoms or moderate or severe pain at presentation were also more likely to experience prolonged zoster-associated pain. Neither time to initiating treatment after rash onset nor sex of patient influenced pain outcome. Advancing age, prodromal symptoms, and acute pain severity at presentation predicted those individuals most at risk of prolonged pain and postherpetic neuralgia. When two or more of these factors were present, the risk of persistent pain was increased. PMID- 9852980 TI - Postherpetic neuralgia: impact of famciclovir, age, rash severity, and acute pain in herpes zoster patients. AB - New and previously reported analyses of the data from a placebo-controlled trial of famciclovir are reviewed in light of recently proposed recommendations for the analysis of pain in herpes zoster trials. The analyses examined the effect of famciclovir treatment on the duration of postherpetic neuralgia (PHN), which was defined as pain persisting after rash healing, pain persisting > 30 days after study enrollment, or pain persisting > 3 months after study enrollment; the baseline characteristics of patients in the famciclovir and placebo groups who developed PHN; the impact of famciclovir treatment on the duration of PHN, while controlling for significant covariates; and the prevalence of PHN at monthly intervals from 30 to 180 days after enrollment. The results of these analyses indicated that greater age, rash severity, and acute pain severity are risk factors for prolonged PHN. In addition, they demonstrated that treatment of acute herpes zoster patients with famciclovir significantly reduces both the duration and prevalence of PHN. PMID- 9852981 TI - Treatment of acute herpes zoster: effect of early (< 48 h) versus late (48-72 h) therapy with acyclovir and valaciclovir on prolonged pain. AB - The efficacy of early versus late treatment with acyclovir and valaciclovir on zoster-associated pain was assessed from two databases (1076 patients) that were compiled from randomized trials. Early treatment was started < 48 h and late treatment was started 48-72 h after the onset of cutaneous herpes zoster. Median times to complete resolution of zoster-associated pain were 28 and 62 days, respectively, for patients (> or = 18 years of age) treated with acyclovir and placebo within 48 h (hazard ratio [HR], 1.68; 95% confidence limit [95% CL], 1.19, 2.38) and 28 and 58 days, respectively, for those treated later (HR, 2.20; 95% CL, 1.03, 4.71). In the valaciclovir versus acyclovir study (in patients > or = 50 years of age), the corresponding figures were 44 and 51 days for patients treated early (HR, 1.28; 95% CL, 1.03, 1.60) and 36 and 48 days for those treated later (HR, 1.40; 95% CL, 1.04, 1.87). Acyclovir significantly shortened the time to complete resolution of zoster-associated pain compared with placebo (and valaciclovir was superior to acyclovir in this regard) even when therapy was delayed up to 72 h after rash onset. PMID- 9852982 TI - Cost effectiveness of newer antiviral agents for herpes zoster: is the evidence spotty? AB - Famciclovir and valaciclovir were approved for use in the treatment of herpes zoster despite controversy over antiviral therapy in zoster due to high costs and uncertain benefits. To explore these issues, a Markov decision model was developed, and the incremental cost effectiveness of antiviral treatment for herpes zoster was estimated using these agents compared with no antiviral therapy. A third-party payer perspective was taken. Sensitivity analyses were performed, modeling differences in antiviral efficacy, postherpetic neuralgia (PHN) risk, and other illness parameters. Treatment of severely symptomatic acute zoster was found reasonable from a cost-effectiveness standpoint in base-case and worst-case scenarios. Treatment of mildly symptomatic acute zoster was more expensive but would likely be considered cost effective in scenarios where PHN risk was higher, PHN duration longer, or antiviral shortening of PHN greater. Further research comparing antiviral efficacy in herpes zoster is needed. PMID- 9852983 TI - Postherpetic neuralgia: the importance of preventing this intractable end-stage disorder. AB - An argument is presented here for postherpetic neuralgia as an intractable end stage disorder for many patients. The exciting possibility of prevention of this disorder by early, aggressive treatment exists; however, the extent to which therapy can be effective is unknown. Early, aggressive treatment of the pain of herpes zoster is, nevertheless, urged, and the options for treatment are discussed. These options include antiviral therapy within the first 72 h, if possible, from the onset of rash or radicular pain and the use of analgesics, including opioids (if necessary), nerve blocks, and early antidepressant therapy. In addition, the extent to which vaccination of older adults will prevent postherpetic neuralgia is unknown but appears to hold promise. PMID- 9852984 TI - Cytokine production in varicella-zoster virus-stimulated cultures of human blood lymphocytes. AB - Estimates of responder cell frequency (RCF) based on limiting dilution analyses are laborious, and alternative means to quantitate cell-mediated immunity to immunogens are desirable. It was shown that levels of interleukin (IL)-2 in the supernatant of varicella-zoster virus-stimulated blood lymphocytes from immune adults peaked at 48 h of culture and correlated partially with estimates of RCF (r = .74, P = .003). Levels of gamma-interferon, IL-4, and IL-10 increased through the first 4 days of culture, and gamma-interferon levels showed some correlation with peak IL-2 levels (r = .48, P = .03). Nevertheless, correlations between levels of these cytokines and RCF did not reach statistically significant levels. PMID- 9852985 TI - A dose-response study of a live attenuated varicella-zoster virus (Oka strain) vaccine administered to adults 55 years of age and older. AB - Decreased cell-mediated immune (CMI) response to varicella-zoster virus (VZV) is correlated with an increased risk of reactivation of latent virus from dorsal root sites, leading to herpes zoster. The cell-mediated and humoral immunogenicity of three concentrations (3200, 8500, and 41,650 pfu/dose) of a live attenuated VZV vaccine (Oka strain; VZV/Oka) was compared with a control pneumococcal polysaccharide vaccine in 200 healthy adults who were > or = 55 years old. Six weeks after vaccination, the VZV-specific CMI response (as measured by stimulation index values and precursor cell frequencies) was enhanced in all VZV/Oka vaccine groups compared with the control group (for all VZV/Oka groups combined vs. controls, tested with VZV crude antigen: stimulation index, P < .001; precursor cell frequency, P < .001). Geometric mean titers of anti-VZV antibodies increased in all VZV/Oka vaccine groups but remained unchanged in the control vaccine group. No dose effect of VZV/Oka vaccine was observed for CMI or humoral responses. PMID- 9852986 TI - Cellular immunity to varicella-zoster virus in patients with major depression. AB - The incidence of herpes zoster increases markedly with advancing age, and this appears to be causally related to an age-dependent decline in varicella-zoster virus (VZV)-specific cellular immunity. Psychologic stress has also been linked to the occurrence of herpes zoster, but the mechanism involved has not been investigated. This study examined the relationship between major depression and VZV-specific cellular immunity by comparing VZV-specific responder cell frequency (RCF) in adults with major depression (n = 11) to that in age- and sex-matched nondepressed controls (n = 11) and in a larger group of nondepressed adults who were > or = 60 years old. VZV-specific RCF in depressed patients was markedly reduced compared with the RCF in matched controls (t = 2.7, P < .02). In fact, the levels of VZV-specific RCF in the depressed patients were comparable in magnitude to the low levels found in adults > or = 60 years of age. These data indicate that major depression is associated with a marked decline in VZV specific cellular immunity. PMID- 9852987 TI - Use of a live attenuated varicella vaccine to boost varicella-specific immune responses in seropositive people 55 years of age and older: duration of booster effect. AB - Varicella-zoster virus (VZV)-specific T cell immunity was measured in 130 persons > or = 55 years of age 6 years after they received a live attenuated VZV vaccine. Circulating T cells, which proliferated in vitro in response to VZV antigen, were enumerated (VZV responder cell frequency assay). Six years after the booster vaccination, the VZV-responding cell frequency (1/61,000 circulating cells) was still significantly (P < .05) improved over the baseline measurements (1/70,000) and appears to have diminished the expected decline in frequency as these vaccinees aged (to 1/86,000). Ten herpes-zoster--like clinical events were recorded. Although the frequency of these events, approximately 1/100 patient years, is within the expected range of such events for this age cohort, the number of lesions was small, there was very little pain, and there was no postherpetic neuralgia. These results support the development of a vaccine to prevent or attenuate herpes zoster. PMID- 9852988 TI - Vitamin C mediated protection on cisplatin induced mutagenicity in mice. AB - In present studies the development of chromosomal aberrations, micronuclei in bone marrow cells and sperm head abnormalities were used as mutagenic bioassay in Swiss albino mice treated with cisplatin alone or ascorbic plus cisplatin. It was noted that in the combined treated hosts the frequency of all the mutagenic parameters were always significantly less than that treated with cisplatin alone. These findings suggest a protective role of ascorbic acid against cisplatin induced mutagenic potentials. Interestingly, in combined treated hosts glutathione (GSH) level in bone marrow cells increased significantly which may suggest a possible mechanism of ascorbic acid mediated protection against cisplatin induced mutagenic potentials in the hosts. PMID- 9852989 TI - Deletion-pattern analysis of alpha-particle and X-ray induced mutations at the HPRT locus of V79 Chinese hamster cells. AB - To investigate the mutagenic mechanisms of low-energy alpha particles V79 Chinese hamster cells were irradiated with 241Am-alpha particles (mean LET of 112 keV/micron). Parallel experiments were performed using 300 kV X-rays. Cell inactivation and mutation induction cross sections were measured. At approximately 20%--survival level, DNA deletions were analysed at the HPRT locus by multiplex-PCR-analysis of all nine exons of 47 alpha-irradiated and 36 background mutants. 92 HPRT- mutants isolated after 300 kV-X-irradiation were analysed similarly for comparison, along with 15 corresponding background mutants. The resulting mutant deletion-pattern distributions were corrected for background mutations. alpha Particles induced a larger fraction of deletions than X-rays. Furthermore, non-contiguous partial deletions were present among the alpha-induced mutants, a type not found after X-irradiation. PMID- 9852990 TI - In situ detection of frameshift mutations in mouse cells. AB - A reporter gene system that allows in situ detection of cells that have suffered a specific frameshift mutation was developed. To construct the reporter gene, the open reading frame of a human placental alkaline phosphatase (PLAP) gene was disrupted by insertion of either 5 or 7 G:C basepairs, which formed mutant alleles carrying 9 and 11 consecutive G residues, respectively. The mutant PLAP genes did not produce alkaline phosphatase activity in cultured mouse cells in transient transfection assays. Several cell lines that contained integrated copies of the mutant PLAP genes were made. Histochemical staining of fixed cells showed that these cell lines contained a small number of cells that expressed PLAP activity and bound antibodies directed against PLAP. Cells carrying the allele with 11 consecutive G residues (G11 allele) acquired PLAP activity at a rate between 2 x 10(-3) and 2 x 10(-4) events per cell per generation, depending on the cell line. Cells carrying the allele with 9 consecutive G residues (G09 allele) acquired PLAP activity at a rate between 2 x 10(-5) and 2 x 10(-6) events per cell per generation, depending on the cell line. Cultures of PLAP+ cells were derived from cell lines carrying PLAP mutant genes. All the cells in these cultures had PLAP activity and bound anti-PLAP antibody. PLAP mRNA levels were the same in cultures where all cells were PLAP+ and in cultures where less than 1% of the cells expressed PLAP activity. DNA sequence analysis of PLAP+ cells showed that the G11 allele reverted by losing one basepair, and the G09 allele reverted by gaining one basepair. PMID- 9852991 TI - Detection of structural chromosome damage in rat interphase cells using region specific fluorescence in situ hybridization probes developed by microdissection. AB - Cytogenetic analysis using fluorescence in situ hybridization (FISH) was employed to detect structural chromosome aberrations in interphase. We generated DNA probes specific for rat chromosome regions 1q11-12, 1q31-35 and 1q51-53 by microdissection and degenerate oligonucleotide-primed PCR. Targeted regions were labeled in unique colors by FISH. Abnormal cells were identified on the basis of alterations in the physical distance between the hybridization signals. To evaluate the ability of these probes to quantify chromosome aberrations in interphase, rats were acutely exposed whole-body to 0, 1, 2, 3 or 4 Gy of 137Cs gamma rays. Eight days later, peripheral blood, bone marrow, lung and pancreas were removed and hybridized with the probes. Multi-color FISH analysis showed dose-responsive frequencies of abnormal interphase nuclei in peripheral blood and bone marrow cells. In lung and pancreas, on the contrary, no increase in the frequency of the abnormal interphases was observed. However, chromosome damage was observed when primary lung cells, obtained from rats irradiated 8 days previously, were cultured for three days. Detection of rearranged signals after in vitro tissue culture was attributed to the movement of chromosome domains that accompanies mitosis. The use of region-specific painting probes appears useful for detecting structural chromosome damage in interphase cells of rat tissues, although further optimization is still needed to improve the method. PMID- 9852993 TI - Micronucleated erythrocytes frequency and radiocesium bioconcentration in pikes (Esox lucius) caught in the Tom River near the nuclear facilities of the Siberian Chemical Complex (Tomsk-7). AB - Between 1957 and 1993, the Siberian Chemical Complex (Tomsk-7) located in the Tomsk region (Russia) regularly discharged radioactive liquid wastes into the Tom River which resulted in an extensive contamination of large territories with long lived radionuclides such as cesium-137 and strontium-90. In the summers of 1996 and 1997, Research Team of Siberian Medical University conducted biodosimetry and cytogenetic monitoring of pikes (Esox lucius) caught in the Tom River at various distances downstream from the Siberian Chemical Complex (SCC) using the micronucleus test and the gamma spectroscopy. Our findings demonstrated that the difference in frequency of micronucleated erythrocytes between the radiation exposed fish caught downstream from the SCC and the controls was statistically significant (P < 0.01). Moreover, we found a good correlation between radiocesium concentration and micronucleated erythrocyte frequency in pikes. It was found that both the micronucleated erythrocyte frequency in pike blood and the level of the pike radiocesium concentration depended on the age of pikes. The micronucleated erythrocyte frequency gradually increased from the 1-year-old pikes to the over 20-year-old pikes. On the other hand, the average level of radiocesium concentration gradually increased from the 1-year-old pikes only up to the 10-year-old pikes. There is no correlation between radiocesium concentration and micronucleated erythrocyte frequency in the over 10-year-old pikes. PMID- 9852992 TI - Mutation of the uracil DNA glycosylase gene detected in glioblastoma. AB - Despite extensive characterization of genetic changes in gliomas, the underlying etiology of these tumors remains largely unknown. Spontaneous DNA damage due to hydrolysis, methylation, and oxidation is a frequent event in the brain. Failure of DNA repair following this damage may contribute to tumorigenesis of gliomas. Uracil DNA glycosylase (UDG), an enzyme which excises uracil from DNA, is an important component of the base excision repair pathway. The sequence of a human homologue of uracil DNA glycosylase gene (UNG) has been recently identified. We performed PCR-based SSCP mutational analysis of UNG in 11 sporadic gliomas (six glioblastomas, two anaplastic astrocytomas, and three oligodendrogliomas) and eight glioblastoma cell lines. One out of six sporadic glioblastomas had a point mutation in exon 3, which resulted in a missense mutation in codon 143. None of the eight glioblastoma cell lines or the five non-glioblastoma sporadic gliomas showed a mutation. Genetic alterations of UNG may play a role in the development of a subset of primary glioblastomas. PMID- 9852994 TI - Mitochondrial DNA rearrangements, including partial duplications, occur in young and old rat tissues. AB - Using polymerase chain reaction (PCR) with back-to-back primers, 85 different mitochondrial DNA (mtDNA) rearrangements, consisting of partial duplications or mini-circles, were detected in brain, liver, and heart tissue from Fischer 344 rats. The regions around the mitochondrial tRNALeu(UUR) gene, the cluster of three tRNA genes [His, Ser(AGY), Leu(UUC)], as well as the region of the displacement loop were analyzed separately with different primer sets. Rearrangements were detected in all regions analyzed in samples taken throughout the animal life span, ranging from 1 day old to 33 months of age (senescent). Two thirds of the rearrangements terminated at short (3-9-bp) direct repeats. Three of the different rearrangements were detected in more than one animal; the most common rearrangement was found in nine different template preparations. Two loci (hot spots) were found to be particularly susceptible to rearrangement, and both were located at sequences that exhibited highly conserved potential for secondary structure formation. The displacement loop region of 10 samples exhibited the presence of multiple tandem duplications ranging between 324 and 449 bp in length. One of these consisted of heterologous, but overlapping, repeating units. Identical PCR protocols were carried out in control experiments using a cloned fragment of mtDNA that encompassed the most common hot spot sequence. The results showed that this fragment did not artifactually generate a rearrangement junction under our PCR conditions and suggested that this sequence does not promote rearrangement mutations in bacteria during the cloning process. PMID- 9852995 TI - Hypermutation as a means to globally re-stabilize the genome following environmental stress. PMID- 9852996 TI - Transient appearance of the mutator phenotype during carcinogenesis as a possible explanation for the lack of mini/microsatellite instability in many late stage tumors. PMID- 9852997 TI - Brussels sprouts: an exceptionally rich source of ambiguity for anticancer strategies. PMID- 9852998 TI - Decreased biliary secretion of proteins and phospholipids by rats with 1,1 dichloroethylene-induced bile canalicular injury. AB - 1,1-Dichloroethylene (DCE, 50 mg/kg) rapidly and selectively injures the bile canalicular membrane of zone 3 hepatocytes. Thus, DCE is of value as a tool to assess the consequences of alterations in canalicular membrane integrity on bile formation. Our objective was to characterize the effects of DCE on the biliary secretion of proteins and phospholipids in freely moving rats. DCE treatment caused a rapid and sustained decrease in total biliary protein output. In contrast, canalicular membrane-localized enzyme activities more slowly increased to 8- to 15-fold in bile from DCE-treated rats. Biliary output of lysosomal enzymes was altered in a biphasic manner. Specifically, there was a transient fivefold increase within 30 min of DCE treatment and then a progressive decrease to approximately 10% basal levels by 4 h. Secretion of phospholipids into bile decreased rapidly in a striking and sustained manner, after DCE. Our findings of diminished lysosomal protein and phospholipid secretion following DCE treatment are consistent with an important role for canalicular membrane integrity in their entry into bile. PMID- 9852999 TI - Benzo(a)pyrene-induced alterations in growth-related gene expression and signaling in precision-cut adult rat liver and kidney slices. AB - Benzo(a)pyrene (BaP) and related aromatic hydrocarbons are suspected carcinogens; however, the molecular basis underlying tumorigenesis remains unclear. To identify acute molecular targets of BaP within the liver and kidney, precision cut slices harvested from naive, adult female Sprague-Dawley rats were challenged with BaP (0.3-30 microM) for 0.5 to 24 h. BaP did not elicit cytotoxicity, as assessed by intracellular K+ and ATP content and histological evaluation over the 24-h period. To determine if molecular signaling pathways were maintained in precision-cut slices, induction of the aryl hydrocarbon receptor (AhR) pathway was assessed following BaP challenge. Induction of cytochrome P450IA1 (P450IA1) mRNA and protein expression was observed in both liver and kidney slices. c-fos and c-Ha-ras gene expression was enhanced in liver, but not kidney, slices by BaP. c-jun mRNA levels were decreased in liver and kidney slices, although the effect was earlier (0.5 h) in liver slices compared to kidney slices. BaP increased the DNA binding of nuclear proteins to the AP-1 consensus recognition element in liver, but decreased DNA binding in kidney slices. In contrast, DNA binding of NF-kappa B was not affected by BaP in either liver or kidney slices. These results suggest that acute BaP challenge is associated with altered expression of several growth-related genes and AP-1 signaling and establish precision-cut slices as a useful in vitro system to investigate the molecular basis of BaP-induced tumorigenesis, including organ-specific differences. PMID- 9853000 TI - Effects of dietary selenium species on Se concentrations in hair, blood, and urine. AB - The effects of the chemical species and concentration of selenium (Se) in diets on the concentrations of Se in hair, blood serum, red blood cells (RBCs), and urine were studied to gain an insight into the toxicological and nutritional significance of different chemical forms of Se. Male Wistar rats were fed an Se deficient diet (Se, less than 0.03 microgram/g) for 3 weeks, and then an Se adequate (Se, 0.2 microgram/g) or Se-excess diet (Se, 2.0 micrograms/g), including seleno-L-methionine (SeMet) or selenite for up to 12 weeks. Hair, blood, and urine specimens were obtained every two weeks, and the concentrations of Se and its distribution in serum and urine on a size-exclusion column were determined. The concentrations of Se in hair, serum, and urine attained constant levels 2 weeks after a change of in the dietary Se concentration irrespective of the chemical species, the levels being dependent on the chemical species and the concentration. Specifically, in hair and serum, selenite gave the lowest constant levels irrespective of the dose, while SeMet resulted in higher levels than selenite in a dose-dependent manner. The two major selenoproteins in serum exhibited comparable concentrations. On the other hand, in urine, the concentration of Se was dependent on the dose but not on the chemical species. The results could be explained by regulated metabolism of selenite, and both nonregulated and regulated aspects of the metabolism of SeMet. PMID- 9853001 TI - Role of angiotensin II, endothelin-1, and nitric oxide in HgCl2-induced acute renal failure. AB - To elucidate the mechanisms underlying the development of HgCl2-induced acute renal failure (ARF), we examined the expression of endothelin (ET)-1, endothelial (e) nitric oxide synthase (NOS) and inducible (i) NOS, and a role of angiotensin II (ANG II) and tumor necrosis factor (TNF) in glomeruli and cortices from rats at 20 h after exposure of HgCl2. Prepro-ET-1 and iNOS mRNA were significantly increased in glomeruli and cortices from rats with HgCl2-induced ARF. However, eNOS mRNA was markedly decreased in glomeruli of rats with HgCl2-induced ARF. Blockade of the action of endogenous ANG II with TCV-116, an ANG II receptor type 1 antagonist, or prior administration of TNF antibody (Ab) neutralizing TNF bioactivity or aminoguanidine, an iNOS inhibitor, substantially suppressed the increase in the expression of prepro-ET-1 or iNOS mRNA seen in rats with HgCl2 induced ARF. Both TCV-116 and TNF Ab had no effects on the expression of eNOS mRNA. The abundance of ET-1, iNOS, and eNOS proteins was paralleled by the magnitude of each mRNA expression. Additionally, the aggravation of blood urea nitrogen and serum Cr observed in rats with HgCl2-induced ARF were significantly ameliorated together with the alleviation of proximal tubule epithelial cell injury when the expression of prepro-ET-1 or iNOS mRNA was blunted by prior administration of TCV-116 or prior injection of TNF Ab or aminoguanidine. These observations indicate that ANG II, ET-1, and NO may play an important role in the progression of HgCl2-induced ARF through the acceleration of proximal tubule epithelial cell injury and the deterioration of glomerular hemodynamics. In HgCl2 induced ARF, the gene expression of ET-1 or iNOS is at least in part up-regulated at the transcription level by endogenous ANG II or TNF. PMID- 9853002 TI - Bile duct obstruction is not a prerequisite for type I biliary epithelial cell hyperplasia. AB - Biliary obstruction, produced by common bile duct ligation or alpha naphthylisothiocyanate (ANIT) treatment in rats, has been associated with the development of type I biliary epithelial cell (BEC) hyperplasia. However, the exact mechanism(s) by which bile duct obstruction lead(s) to this proliferative lesion are not clear. The present studies were designed to determine if cholestasis, in the absence of biliary obstruction, would result in type I BEC hyperplasia. Male Sprague-Dawley rats were given a single oral dose of 150 mg/kg ANIT or i.v. doses of estradiol glucuronide (E2-17G; 21 mumol/kg/h for 48 h) to produce obstructive and non-obstructive cholestasis, respectively. E2-17G treatment resulted in cholestasis that was comparable in extent and duration to that observed following ANIT treatment. E2-17G and ANIT treatments produced comparable increases in serum bile acids (55- to 60-fold) and activities of ALT (36- to 38-fold), ALP (4- to 5-fold), and 5'-nucleotidase (7- to 11-fold), respectively, compared to controls. Both ANIT and E2-17G also increased serum bilirubin concentrations. ANIT treatment resulted in significant increases in biliary glucose concentrations that were associated with BEC damage/necrosis and obstruction of the bile duct lumen. Conversely, no evidence of BEC damage was observed in E2-17G-treated rats. Nonetheless, BEC hyperplasia was observed in the majority of rats following treatment with either ANIT or E2-17G, assessed by light microscopy and by BrdU immunohistochemistry. These data indicate that E2 17G treatment produces nonobstructive cholestasis and type I BEC hyperplasia, suggesting that biliary obstruction is not a prerequisite for type I BEC hyperplasia in rats. Differences in the time of onset of hyperplasia were observed: hyperplasia was noted immediately following 48 h of E2-17G-induced cholestasis but occurred several days after ANIT-induced cholestasis had subsided. Since the magnitude/duration of cholestasis was similar in the two models but the temporal association between cholestasis and type I BEC hyperplasia were different, these data suggest that the proliferative stimulus may be different in the two models and that E2-17G-induced type I BEC hyperplasia may not be attributed solely to cholestasis. PMID- 9853003 TI - A human physiologically based pharmacokinetic model for trichloroethylene and its metabolites, trichloroacetic acid and free trichloroethanol. AB - Nine male and eight female healthy volunteers were exposed to 50 or 100 ppm trichloroethylene vapors for 4 h. Blood, urine, and exhaled breath samples were collected for development of a physiologically based pharmacokinetic (PBPK) model for trichloroethylene and its two major P450-mediated metabolites, trichloroacetic acid and free trichloroethanol. Blood and urine were analyzed for trichloroethylene, chloral hydrate, free trichloroethanol and trichloroethanol glucuronide, and trichloroacetic acid. Plasma was analyzed for dichloroacetic acid. Trichloroethylene was also measured in exhaled breath samples. Trichloroethylene, free trichloroethanol, and trichloroacetic acid were found in blood samples of all volunteers and only trace amounts of dichloroacetic acid (4 12 ppb) were found in plasma samples from a few volunteers. Trichloroethanol glucuronide and trichloroacetic acid were found in urine of all volunteers. No chloral hydrate was detected in the volunteers. Gender-specific PBPK models were developed with fitted urinary rate constant values for each individual trichloroethylene exposure to describe urinary excretion of trichloroethanol glucuronide and trichloroacetic acid. Individual urinary excretion rate constants were necessary to account for the variability in the measured cumulative amount of metabolites excreted in the urine. However, the average amount of trichloroacetic acid and trichloroethanol glucuronide excreted in urine for each gender was predicted using mean urinary excretion rate constant values for each sex. A four-compartment physiological flow model was used for the metabolites (lung, liver, kidney, and body) and a six-compartment physiological flow model was used for trichloroethylene (lung, liver, kidney, fat, and slowly and rapidly perfused tissues). Metabolic capacity (Vmaxc) for oxidation of trichloroethylene was estimated to be 4 mg/kg/h in males and 5 mg/kg/h in females. Metabolized trichloroethylene was assumed to be converted to either free trichloroethanol (90%) or trichloroacetic acid (10%). Free trichloroethanol was glucuronidated forming trichloroethanol glucuronide or converted to trichloroacetic acid via back conversion of trichloroethanol to chloral (trichloroacetaldehyde). Trichloroethanol glucuronide and trichloroacetic acid were then excreted in urine. Gender-related pharmacokinetic differences in the uptake and metabolism of trichloroethylene were minor, but apparent. In general, the PBPK models for the male and female volunteers provided adequate predictions of the uptake of trichloroethylene and distribution of trichloroethylene and its metabolites, trichloroacetic acid and free trichloroethanol. The PBPK models for males and females consistently overpredicted exhaled breath concentrations of trichloroethylene immediately following the TCE exposure for a 2- to 4-h period. Further research is needed to better understand the biological determinants responsible for the observed variability in urinary excretion of trichloroethanol glucuronide and trichloroacetic acid and the metabolic pathway resulting in formation of dichloroacetic acid. PMID- 9853004 TI - Verapamil protection against mercuric chloride-induced renal glomerular injury in rats. AB - We have examined the effects of the calcium channel blocker verapamil on the renal glomerular structural damage produced by mercuric chloride in rats. Verapamil (75 micrograms/kg body wt iv) was administered 30 min prior to mercuric chloride injection (HgCl2, 5 mg/kg body wt sc). Verapamil prevented the glomerular proteinuria observed in HgCl2-treated rats. Isolated glomeruli from mercury-treated rats 1 h after injection presented a diminished cross-sectional area as compared with control glomeruli (control [micron2], 26,310 +/- 2545; HgCl2 [micron2], 18,474 +/- 1828) and increased glomerular calcium content (control, 23 +/- 6 nmol/mg protein; HgCl2, 43 +/- 7 nmol/mg protein). Verapamil pretreatment prevented glomerular cross-sectional area (GCSA) diminution and glomerular calcium content rise (GCSA [micron2] Vp + Hg, 28,281 +/- 4654, Ca2+ [nmol/mg protein] Vp + Hg, 18 +/- 5). Renal sections prepared for immunohistochemical detection and histochemical analysis showed increased deposits of fibronectin and lipids and enhanced cellularity in glomerular structures from HgCl2-treated rats. Renal sections from animals pretreated with verapamil showed fibronectin and lipid contents not different from control sections and their histological studies did not show any changes when compared with control. Verapamil pretreatment also protected glomeruli from enhanced leukocyte content (myeloperoxidase activity/mg protein): control, 59 +/- 7; HgCl2, 134 +/- 10; Vp + Hg, 79 +/- 11). HgCl2 also contracts GCSA in vitro; Vp prevented this GCSA diminution. The results described in this study indicate that mercuric chloride nephrotoxicity may be associated not only with changes in renal glomerular haemodynamics, but also with a direct effect on glomerular cells. PMID- 9853005 TI - Enzymatic methylation of arsenic compounds. VI. Characterization of hamster liver arsenite and methylarsonic acid methyltransferase activities in vitro. AB - Methylation of inorganic arsenic to methylarsonic acid (MMA) and dimethylarsinic acid (DMA) has been considered to be the major pathway of inorganic arsenic biotransformation and detoxification. Comparative studies, in vivo, have demonstrated variation in the abilities of animals to methylate inorganic arsenic. We propose that the rate of inorganic arsenite methylation may be one of the factors responsible for observed species variation. Arsenite and MMA methyltransferases of Golden Syrian hamster liver have been partially purified 40 and 67-fold, respectively. The monothiol L-cysteine promotes greater activities, in vitro, of these enzymes than similar concentrations of either glutathione or dithiothreitol. The pH optima of the partially purified arsenite and MMA methyltransferase activities are 7.6 and 8.0, respectively. Both activities display classic Michaelis-Menten enzyme kinetics. The K(m) and Vmax of hamster liver arsenite methyltransferase are 1.79 x 10(-6) M and 0.022 pmol/mg protein/60 min, respectively. Hamster liver MMA methyltransferase has K(m) and Vmax values of 7.98 x 10(-4) M and 0.007 pmol/mg protein/60 min, respectively. A similar kinetic relationship of these activities is also observed in the liver of the rabbit, which, like the hamster, excretes higher amounts of MMA than most other species studied. The higher K(m) and lower Vmax of MMA methyltransferase, compared to arsenite methyltransferase, measured in these two species suggests that MMA may be produced at a rate higher than it can be subsequently methylated to DMA, thereby allowing MMA to accumulate and be excreted. PMID- 9853006 TI - In vitro to in vivo extrapolation for trichloroethylene metabolism in humans. AB - The use of in vitro systems in the assessment of xenobiotic metabolism has distinct advantages and disadvantages. While isolated hepatocytes and microsomes prepared from human liver may be used to generate data for comparisons among species and in vitro systems, such comparisons are generally performed on the basis of microsomal protein or million (viable) hepatocytes. Recently, in vitro data have been investigated for their value as quantitative predictors of in vivo metabolic capacity. Because of the existence of large amounts of trichloroethylene (TRI) data in the human, we have examined the metabolism of TRI as a case study in the development of a method to compare metabolism across species using in vitro systems and for extrapolation of metabolic rates from in vitro to in vivo. TRI is well metabolized by human hepatocytes in culture with a K(m) of 266 +/- 202 ppm (mean +/- SD) in headspace and a Vmax of 16.1 +/- 12.9 nmol/h/10(6) viable hepatocytes. We determined that human liver contains approximately 116 x 10(6) hepatocytes and 20.8 mg microsomal protein/g, based on DNA recovery and glucose-6-phosphatase activity, respectively. Thus, the microsomal protein content of hepatocytes is 179 micrograms microsomal protein/10(6) isolated hepatocytes. The microsomal apparent Vmax value of 1589 pmol/min/mg microsomal protein extrapolates to 17.07 nmol/h/10(6) hepatocytes. The combination of protein recovery and metabolic rate predicted a Vmax of approximately 1400 nmol/h/g human liver, which, when extrapolated and incorporated into an existing physiologically based pharmacokinetic (PBPK) model for TRI, slightly underpredicted TRI metabolism in the intact human. The quantitation, extrapolation, and inclusion of extrahepatic and cytochrome P450 (CYP)-independent TRI metabolism may increase the predictive value of this approach. PMID- 9853007 TI - Expression and activity of urokinase and its receptor in endothelial and pulmonary epithelial cells exposed to asbestos. AB - An elongated endothelial cell phenotype, which demonstrated increased ICAM-1 dependent neutrophil adherence, was induced when these cells were exposed to noncytotoxic concentrations of asbestos (Treadwell et al., Toxicol. Appl. Pharmacol. 139, 62-70, 1996). The present study examined mechanisms underlying this phenotypic change by investigating the effects of asbestos on transcription factor activation and expression of urokinase-type plasminogen activator (uPA) and its receptor uPAR. In situ zymography was used to compare the effects of these fibers on the activity of uPA. Cultures incubated with chrysotile or crocidolite asbestos, but not refractory ceramic fiber 1 (RCF-1), demonstrate localized cleavage of plasminogen, which was inhibited by amiloride. Immunocytochemistry showed that chrysotile-stimulated uPA activity was associated with a time-dependent augmentation of uPAR protein levels. RT-PCR analysis was used to investigate molecular mechanisms for these increases. Chrysotile asbestos, but not RCF-1, increased endothelial cell uPA message, relative to changes in beta-actin mRNA. This response to asbestos was not limited to endothelial cells, since both uPA and uPAR mRNA levels increase in human bronchial epithelial BEAS-2B cells exposed to chrysotile fibers. Finally, both types of asbestos, but not RCF-1, increased nuclear levels of nuclear factor kappaB (NF-kappa B), a transcription factor common to increased expression of ICAM-1 and uPA. These data demonstrate that asbestos caused fiber-specific activation of endothelial and pulmonary epithelial cells, resulting in phenotypes capable of facilitating tissue remodeling. PMID- 9853008 TI - Inhibition of poly(ADP-ribose) polymerase rescues human T lymphocytes from methylmercury-induced apoptosis. AB - The objective of this investigation was to determine the role of poly(ADP-ribose) polymerase (PARP) in methylmercuric chloride (MeHgCl)-induced T-cell apoptosis. Following exposure of human T-cells to 2.5 microM MeHgCl, we observed PARP activation within 45 min. Maximal activation was observed at 90 min after MeHgCl treatment; thereafter, PARP activity declined. The loss in enzyme activity was coincidental with the cleavage of 116-kDa intact PARP protein to an 85-kDa fragment. To address the relationship between PARP activation and induction of apoptosis, we first examined the redox status of T cells treated with MeHgCl. We found that exposure of T cells to low concentrations of this toxicant resulted in decreased levels of reduced pyridine nucleotides and an increase in the relative amounts of oxidized flavoproteins. Thus, the possibility exists that activation of PARP leads to NAD+ depletion and thereby alters mitochondrial redox status. To determine if PARP activation is indeed part of the proapoptotic (destructive) response or a component of the antiapoptotic (protective) response, we employed two inhibitors: 3-aminobenzamide and nicotinamide. Pretreatment of T cells with these inhibitors protected cells from MeHgCl-induced apoptosis; this was seen as a reduction in the uptake of Hoechst 33258 and DNA fragmentation. Moreover, these inhibitors blocked MeHgCl-induced oxidative stress as evidenced by a reduction in reactive oxygen species (ROS) generation. These agents, however, failed to block MeHgCl-dependent decline in mitochondrial transmembrane potential (delta psi m). We conclude that PARP activation leads to proapoptotic events that contribute to MeHgCl-induced cell death. PMID- 9853010 TI - Renal nerve stimulation restores tubuloglomerular feedback after acute renal denervation. AB - Renal nerves play an important role in the setting of the sensitivity of the tubuloglomerular feedback (TGF) mechanism. We recently reported a time-dependent resetting of TGF to a lower sensitivity 3-4 h after acute unilateral renal denervation (aDNX). This effect persisted after 1 week, but was then less pronounced. To determine whether normal TGF sensitivity could be restored in aDNX kidneys by low-frequency renal nerve stimulation (RNS), the following experiments were performed. Rats with aDNX were prepared for micropuncture. In one experimental group proximal tubular free flow (Pt) and stop flow pressures (Psf) were measured during RNS at frequencies of 2, 4 and 6 Hz. In another series of experiments the TGF sensitivity was evaluated from the Psf responses at different loop perfusion rates after 20 min of RNS at a frequency of 2 Hz. The maximal drop in Psf (delta Psf) and the tubular flow rate at which half the maximal response in delta Psf was observed (turning point, TP), were recorded. At RNS frequencies of 2, 4 and 6 Hz, Pt decreased from the control level of 14.1 +/- 0.8-13.1 +/- 1.0, 12.4 +/- 1.1 and 11.2 +/- 0.8 mmHg (decrease 21%, P < 0.05), respectively, while at zero perfusion and during RNS at 2 and 4 Hz Psf decreased from 42.5 +/- 1.6 to 38.2 +/- 1.4 and 32.8 +/- 4.3 mmHg (decrease 23%, P < 0.05), respectively. The TGF characteristics were found to be reset from the normal sensitivity with TP of 19.0 +/- 1.1 nL min-1 and delta Psf of 8.7 +/- 0.9 mmHg to TP of 28.3 +/- 2.4 nL min-1 (increase 49%, P < 0.05) and delta Psf of 5.8 +/- 1.2 mmHg (decrease 33%) after aDNX. After 20 min of RNS at 2 Hz TP was normalized and delta Psf was 33% higher. Thus the present findings indicate that the resetting of the TGF sensitivity that occurred 2-3 h after aDNX could be partially restored by 20 min of RNS at a frequency of 2 Hz. These results imply that renal nerves have an important impact on the setting of the sensitivity of the TGF mechanism. PMID- 9853009 TI - Carbaryl, a carbamate insecticide, is a ligand for the hepatic Ah (dioxin) receptor. AB - The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that mediates many of the biological and toxicological actions of a variety of hydrophobic natural and synthetic chemicals, including the environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin). Induction of CYP1A1 gene expression is one such response that is known to be regulated by the AhR complex. It was recently reported (Ledirac et al., Toxicol. Appl. Pharmacol. 144, 177-182, 1997) that, although carbaryl, a carbamate insecticide, can induce AhR-dependent expression of CYP1A1, it was not an AhR ligand. Since this apparent ligand-independent activation of the AhR is difficult to reconcile given what is known about the mechanism of AhR action, we have examined the ability of carbaryl to stimulate the AhR signaling pathway. Not only was dioxin responsive element driven luciferase gene expression induced by carbaryl in stably transfected mouse, rat, guinea pig, and human cells, gel retardation analysis revealed that carbaryl stimulated AhR transformation and DNA binding in vitro and in cells in culture. Dose-response experiments revealed that carbaryl was 300,000-fold less potent that the prototypical inducer, TCDD, in both inducing luciferase gene expression and stimulating AhR transformation and DNA binding in vitro, suggesting that carbaryl itself was the inducing agent. The identification of carbaryl as an AhR ligand was demonstrated by its ability to competitively inhibit [3H]-TCDD to the guinea pig hepatic cytosolic AhR. Our results confirm that carbaryl is both a weak AhR ligand and inducer of AhR-dependent gene expression and argue against its proposed ligand-independent mechanism of AhR activation. PMID- 9853011 TI - Autonomic nervous influence of the female guinea-pig urinary bladder during the oestrus cycle. AB - The mammalian urinary bladder receives dual innervation. The excitatory innervation is considered to be partly cholinergic and partly mediated via NANC receptors. Several (co-)transmitters have been suggested. The adrenergic inhibitory innervation is mediated via alpha- and beta-receptors. Female sex hormones could change autonomic influence of urogenital organs. It was considered to be of interest to characterize the spontaneous and nerve stimulation-induced muscular activity in the urinary bladder of the female guinea-pig during the oestrus cycle. Both the spontaneous activity and nerve-induced activity varied according to the hormonal status of the animal. An alpha-adrenergic inhibitory influence was identified. It was further confirmed that the excitatory innervation could not be blocked by the cholinergic antagonist scopolamine, while alpha-beta-methylene ATP partly inhibited nerve stimulation-induced smooth muscle response, most prominent at cycle day 6. Indomethacin did not impair spontaneous activity or nerve stimulation-induced activity. Nitric oxide reduced nerve stimulation-induced responses on cycle day 12. Imperative urinary bladder contractions are reported to diminish after oestrogen use and in the female a hormonal effect of the nervous influence on the urinary bladder smooth muscle is suggested. PMID- 9853012 TI - Changes in peak oxygen uptake and plasma volume in fit and unfit subjects following exposure to a simulation of microgravity. AB - To test the hypothesis that the magnitude of reduction in plasma volume and work capacity following exposure to simulated microgravity is dependent on the initial level of aerobic fitness, peak oxygen uptake (VO2peak) was measured in a group of physically fit subjects and compared with VO2peak in a group of relatively unfit subjects before and after 10 days of continuous 6 degrees head-down tilt (HDT). Ten fit subjects (40 +/- 2 year) with mean +/- SE VO2peak = 48.9 +/- 1.7 mL kg-1 min-1 were matched for age, height, and lean body weight with 10 unfit subjects (VO2peak = 37.7 +/- 1.6 mL kg-1 min-1). Before and after HDT, plasma, blood, and red cell volumes and body composition were measured and all subjects underwent a graded supine cycle ergometer test to determine VO2peak period needed. Reduced VO2peak in fit subjects (-16.2%) was greater than that of unfit subjects (-6.1%). Similarly, reductions in plasma (-18.3%) and blood volumes (-16.0%) in fit subjects were larger than those of unfit subjects (blood volume = -5.6%; plasma volume = -6.6%). Reduced plasma volume was associated with greater negative body fluid balance during the initial 24 h of HDT in the fit group (912 +/- 154 mL) compared with unfit subjects (453 +/- 200 mL). The percentage change for VO2peak correlated with percentage change in plasma volume (r = +0.79). Following exposure to simulated microgravity, fit subjects demonstrated larger reductions in VO2peak than unfit subjects which was associated with larger reductions in plasma and blood volume. These data suggest that the magnitude of physical deconditioning induced by exposure to microgravity without intervention of countermeasures was influenced by the initial fitness of the subjects. PMID- 9853013 TI - Cardiovascular and muscular adaptations to combined endurance and strength training in elderly women. AB - Twenty-one women aged 60-75 years were examined to determine whether combined endurance and strength training resulted in greater increase in peak oxygen consumption, sub-maximal time to fatigue, cardiac output, stroke volume, and leg extension load when compared to endurance training alone. Subjects in both the endurance training (E) and endurance and strength (E & S) groups trained 3 days a week, for 12 weeks, at an intensity of 70-80% Vo2 peak for 30 min on a cycle ergometer. Subjects in the E & S groups also used resistance equipment to train the knee extensors. The workload for resistance training was based on an initial assessment of 10 repetitions maximum (10 RM), with 80% of that value used for training, three times weekly. Peak oxygen consumption increased to an average of 24.8 and 29.9% in the E and E & S groups, respectively, with no difference between groups. Subjects in the E & S and E groups significantly increased sub maximal endurance time by 396 and 165%, respectively. Cardiac output, stroke volume, and arteriovenous oxygen difference at 80% peak VO2 were unchanged by either of the training methods. A needle biopsy was taken from the vastus lateralis before and after 12 weeks of training. Chi-square analysis of fibre area data showed an increase in the frequency of larger type I fibres in the post training data from the E & S group, but an increase in the frequency of smaller fibres in the E group post-training; however, mean fibre area was not significantly changed by training. These data suggest that greater improvements in sub-maximal time to fatigue and strength is achieved when resistance training is added to an aerobic training programme in healthy elderly women. PMID- 9853014 TI - Contribution of pH, diprotonated phosphate and potassium for the reflex increase in blood pressure during handgrip. AB - The relative importance of pH, diprotonated phosphate (H2PO4-) and potassium (K+) for the reflex increase in mean arterial pressure (MAP) during exercise was evaluated in seven subjects during rhythmic handgrip at 15 and 30% maximal voluntary contraction (MVC), followed by post-exercise muscle ischaemia (PEMI). During 15% MVC, MAP rose from 92 +/- 1 to 103 +/- 2 mmHg, [K+] from 4.1 +/- 0.1 to 5.1 +/- 0.1 mmol L-1, while the intracellular (7.00 +/- 0.01 to 6.80 +/- 0.06) and venous pH fell (7.39 +/- 0.01 to 7.30 +/- 0.01) (P < 0.05). The intracellular [H2PO4-] increased 8.4 +/- 2 mmol kg-1 and the venous [H2PO4-] from 0.14 +/- 0.01 to 0.16 +/- 0.01 mmol L-1 (P < 0.05). During PEMI, MAP remained elevated along with the intracellular [H2PO4-] as well as a low intracellular and venous pH. However, venous [K+] and [H2PO4-] returned to the level at rest. During 30% MVC handgrip, MAP rose to 130 +/- 3 mmHg, [K+] to 5.8 +/- 0.2 mmol L-1, the intracellular and extracellular [H2PO4-] by 20 +/- 5 mmol kg-1 and to 0.20 +/- 0.02 mmol L-1, respectively, while the intracellular (6.33 +/- 0.06) and venous pH fell (7.23 +/- 0.02) (P < 0.05). During post-exercise muscle ischaemia all variables remained close to the exercise levels. Analysis of each variable as a predictor of blood pressure indicated that only the intracellular pH and diprotonated phosphate were linked to the reflex elevation of blood pressure during handgrip. PMID- 9853015 TI - Albumin infusion in humans does not model exercise induced hypervolaemia after 24 hours. AB - We rapidly infused 234 +/- 3 mL of 5% human serum albumin in eight men while measuring haematocrit, haemoglobin concentration, plasma volume (PV), albumin concentration, total protein concentration, osmolality, sodium concentration, renin activity, aldosterone concentration, and atrial natriuretic peptide concentration to test the hypotheses that plasma volume expansion and plasma albumin content expansion will not persist for 24 h. Plasma volume and albumin content were expanded for the first 6 h after infusion (44.3 +/- 1.9-47.2 +/- 2.0 mL kg-1 and 1.9 +/- 0.1-2.1 +/- 0.1 g kg-1 at pre-infusion and 1 h, respectively, P < 0.05), but by 24 h plasma volume and albumin content decreased significantly from 1 h post-infusion and were not different from pre-infusion (44.8 +/- 1.9 mL kg-1 and 1.9 +/- 0.1 g kg-1, respectively). Plasma aldosterone concentration showed a significant effect of time over the 24 h after infusion (P < 0.05), and showed a trend to decrease at 2 h after infusion (167.6 +/- 32.5(-1) 06.2 +/- 13.4 pg mL-1, P = 0.07). These data demonstrate that a 6.8% expansion of plasma volume and 10.5% expansion of plasma albumin content by infusion does not remain in the vascular space for 24 h and suggest a redistribution occurs between the intravascular space and interstitial fluid space. PMID- 9853016 TI - Alpha 2-adrenoceptor activation may trigger the increased production of endothelium-derived nitric oxide in skeletal muscle during acute haemorrhage. AB - Our previous studies indicated that acute haemorrhage leads to a pronounced increase in the release of endothelium-derived nitric oxide (EDNO) graded in relation to the magnitude of the blood loss. The EDNO-induced vasodilatation, confined selectively to the arterial 'feeder' vessels, attenuates the concomitant reflex adrenergic constriction and thereby prevents deleterious reduction of blood flow. The present study aimed at investigating whether the reflex release of blood-borne catecholamines might trigger this EDNO release via activation of endothelial alpha 2-adrenoceptors. The study was performed on the sympathectomized vascular bed of cat skeletal muscle with a technique permitting quantitative recordings of resistance (tone) in consecutive vascular sections. Selection alpha 2-adrenoceptor blockade with idazoxan applied at steady state vasoconstriction after a 35% blood loss evoked an initial generalized dilator response (attributable to inhibition of post-synaptic smooth muscle alpha 2 adrenoceptors), followed by a constrictor response selectively in the arterial feeder vessels, the latter compatible with the hypothesis of reduced EDNO release by alpha 2-adrenoceptor blockade. More direct evidence for the hypothesis was obtained from studies of the vascular response to EDNO blockade (L-NAME) after haemorrhage in the presence and absence of alpha 2-adrenoceptor blockade. The constrictor response to EDNO blockade, which is a measure of the pre-existing EDNO dilator influence (EDNO production), was significantly smaller (P < 0.01) in the presence than absence of alpha 2-adrenoceptor blockade. The results indicate that blood-borne catecholamines, via activation of endothelial alpha 2 adrenoceptors, trigger the increase in the EDNO release in acute haemorrhage, implying a functionally important negative feedback in the integrated control of vascular tone in bleeding. PMID- 9853017 TI - Initiation of increase in muscle sympathetic nerve activity delay during maximal voluntary contraction. AB - To investigate the effects of maximal voluntary exercise on sympathetic nerve activity, contraction force and muscle sympathetic nerve activity (MSNA) were recorded during maximal (MVG) and submaximal voluntary isometric handgrip (SVG) for 2 min in eight healthy subjects. MSNA was determined by a microneurographic technique, and handgrip force, heart rate (HR) and arterial blood pressure (ABP) were measured by a non-invasive method during exercise. Grip force decayed rapidly to 58% of maximal grip force (MGF) at 10 s after commencement of exercise and was almost constant (approximately 30% of MGF) 40 s after exercise. MSNA increase was delayed by 20 s during MVG, followed by a gradual increase. HR was elevated immediately after onset of exercise, while mean ABP rise showed a 20 s lag from initiation of MVG exercise. During SVG increases in MSNA, HR and mean ABP were delayed by 50, 40 and 20 s, respectively, relative to commencement of exercise. Thereafter, these parameters increased time-dependently. These results suggested that the MSNA increase during MVG may be predominantly because of the metaboreflex. PMID- 9853018 TI - Fibre-type specific expression of fast and slow essential myosin light chain mRNAs in trained human skeletal muscles. AB - The fibre-type specific expression patterns of fast and slow isoforms of essential (alkali) myosin light chains (ELC) was analysed in trained, untrained and pathological human muscles. Biopsies from m. vastus lateralis of moderately trained and untrained persons, as well as highly trained endurance and strength athletes were analysed, by in situ hybridization, for the expression of the 'fast' ELC 1f/3f and the 'slow' ELC 1 sb. We wanted to investigate if changes in the fibre-type specific ELC mRNA pattern could be used as markers for training adaptation, especially, if the mRNA of the slow ELC 1sb isoform would appear in type IIA fibres as a result of endurance training (Baumann et al. 1987). We found the fast/slow ELC expression patterns in the fibre types to be remarkably stable. Physiological stress, even high training loads, did not affect it. No IIA fibres expressing ELC 1sb mRNA were found. They could be detected, however, in pathological muscle samples, where fast/slow ELC patterns not found in normal muscles were frequent. Our data suggest that in healthy muscles, only a subset of the theoretically possible combinations of myosin heavy and light chain isoforms are expressed at the level of their mRNAs. PMID- 9853019 TI - Beta-adrenergic stimulation of cellular K+ uptake in rat distal colon. AB - We recently demonstrated that the ratio between colonic K+ absorptive and K+ secretive pathways was higher in infant than in adult rats. To test the hypothesis that hormones selectively affect these pathways during ontogeny we examined the effect of adrenergic agonists on cellular K+ uptake in distal colon from infant (10-day-old) and adult (50-day-old) rats. Here we describe that adrenaline (10(-5) M) increased total and ouabain-insensitive 86Rb uptake in both age groups, but it did not affect ouabain-sensitive 86Rb uptake. This stimulation was more pronounced in adult than in infant rats. The effect of adrenaline was mediated via beta-adrenergic receptors. Incubation in vitro with beta-agonist, isoproterenol, stimulated SCH-28080-sensitive, i.e. H+, K(+)-ATPase-dependent, 86Rb uptake in adult but not in infant rats. The threshold dose of beta-agonist was at 10(-7) M, and the maximal activation was observed at 10(-5) M. In vivo inhibition of beta-adrenergic system with propranolol caused a significant decrease in H+, K(+)-ATPase-dependent 86Rb uptake in infant but not in adult colon. In conclusion, this study suggests that the higher colonic K+ absorption in infant rats may be as a result of a selective beta-adrenergic up-regulation leading to stimulation of the apical H+, K(+)-ATPase. PMID- 9853020 TI - Intraduodenal neuropeptide levels, but not plasma levels, vary in a cyclic fashion with the migrating motor complex. AB - The neurohormonal control of the migrating motor complex (MMC) is not fully understood. The hypothesis of the present study was that neuropeptide levels might vary with the different phases of the MMC and that a similar variation might be found in the secretions of the gastrointestinal tract. Thus, plasma and intraduodenal concentrations of vasoactive intestinal peptide (VIP), somatostatin (SOM), substance P (SP) and neurokinin A (NKA) were determined by radioimmunoassay every 10 min during two complete MMC cycles in eight male subjects. For comparison, plasma motilin (MOT) concentrations were measured. Plasma concentrations of MOT (mean peak value +/- SEM; 39 +/- 6 pmol L-1), but none of the neuropeptides studied, showed a cyclic variation in plasma with the different phases of the MMC. Peak intraduodenal concentrations of VIP (79 +/- 23 pmol L-1), SOM (2437 +/- 432 pmol L-1) and SP (718 +/- 326 pmol L-1) occurred at or at the time point before the onset of phase III of the MMC. No such correlation was observed for NKA. These results demonstrate that intraduodenal but not plasma concentrations of the neuropeptides VIP, SOM and SP show an association with phase III of the MMC. The biological relevance of this finding is yet unclear, but the results raise the possibility that gut neuropeptides may regulate fasting motility through a luminal release. PMID- 9853021 TI - Glucose consumption by various tissues in pregnant rats: effects of a 6-day euglycaemic hyperinsulinaemic clamp. AB - In the course of pregnancy maternal tissues become increasingly more insensitive to insulin. As 6 days of euglycaemic hyperinsulinaemic clamping, from day 8 until 14 of gestation, ameliorates total glucose consumption, we analysed the contribution of individual tissues in this phenomenon. We measured not only glucose consumption, but also concentrations of the glucose transporter protein GLUT4 in selected tissues. On day 15 of pregnancy in saline-infused (pregnant control) rats, 18F-fluoro-2-deoxy-D-glucose (FDG) consumption, as measured by positron emission tomography, as well as GLUT4 content were diminished in heart (P < 0.05), and in brown (P < 0.05) and white adipose tissues (P < 0.05) when compared with non-pregnant controls. During clamping, on day 13 of the experiments, both in pregnant and non-pregnant rats FDG consumption was increased in liver (P < 0.05), skeletal muscle (P < 0.05), brown (P < 0.05) and white adipose tissues (P < 0.05) when compared with saline-infused controls. In both the reproductive conditions, only in white adipose tissue was this increased FDG consumption associated with increased GLUT4 levels (P < 0.05); GLUT4 content of m. gastrocnemius, m. soleus, heart and brown adipose tissue was unaffected by clamping. In non-pregnant rats, 24 h after clamping ceased, FDG consumption was diminished in heart compared with control non-pregnant rats (P < 0.05). In pregnant rats, however, 24 h after clamping (i.e. day 15 of pregnancy) both FDG consumption (P < 0.05) and GLUT4 content (P < 0.05) were still increased in white adipose tissue only, when compared with saline-infused pregnant rats on the same day of gestation. It is suggested that hyperinsulinaemic euglycaemic clamping ameliorates insulin action halfway pregnancy, in particular by stimulation of glucose consumption and GLUT4 protein content in white adipose tissue. PMID- 9853022 TI - GABA and human spermatozoa: characterization and regulation of GABA transport proteins. PMID- 9853023 TI - [Human herpesvirus 8 ( HHV-8 ): I. Characteristics and epidemiology]. AB - HHV8 is a new herpesvirus recently identified in the Kaposi's sarcoma lesions, and initially named Kaposi's sarcoma-associated herpesvirus. It is a member of gamma-2 herpesvirus family and it shows a number of homologies with the Epstein Barr virus and the herpesvirus saimiri. HHV8 genome also codes for several proteins which are homologous to cellular proteins and could disturb the regulation mechanisms of cellular proliferation and apoptosis. This is the case for a viral IL6, an antiapoptotic factor homologous to Bcl2, a viral cyclin, a member of the IRF family (interferon regulatory factors) and a G-protein-coupled receptor homologous to the IL8 receptor. Seroprevalence studies showed that HHV8 infection was not ubiquitous but rather limited to some geographic areas (Italy, Greece, Africa), and to some populations of homosexual and bisexual individuals with sexually transmitted diseases. To date, several lines of epidemiologic evidence suggest that this virus is sexually transmitted, although other routes of transmission cannot be excluded. PMID- 9853024 TI - [Apolipoprotein E and its alleles in healthy subjects and in atherosclerosis]. AB - Apolipoprotein E (apo E) plays a central role in lipoprotein metabolism as a ligand for LDL-, VLDL- and remnant receptors. It is mainly synthesized by hepatocytes and also by monocytes-macrophages. There are three common codominant isoforms E2, E3 and E4. Apo E2 binds defectively to LDL- and to remnant receptors. Apo E alleles frequency varies from population to population around the world. In Europe, E4 allele frequency increases from south to north along the cardiovascular disease frequency gradient. On the other hand, the association between E2 allele and these diseases remains to be proved except for type III hyperlipoproteinemia. Apo E4 role in atherosclerosis could be explained, at least in part, by its high solubility in apo B lipoproteins. The average cholesterolemia of E4/E3 subjects is higher than E3/E3 subjects and E3/E3 subjects' cholesterolemia is higher than in E3/E2 subjects, probably because of a faster uptake of chylomicrons and VLDL remnants in E4/E3 subjects. The E4 allele also seems to play a role not only in diet response in combination with other genetic factors such as the Eco RI polymorphism of apo B but also in hypolipemic treatment. Apo E plays also a role in triglyceridemia control in association with other genetic and environmental factors such as lipoprotein lipase, apo B, LDL receptor and diet. Models of apo E knockout mice suggest a protective effect of monocyte-macrophageapo E in the vessel wall and HDL-apo E probably induces a high cholesterol efflux from macrophages. This effect counteracts the proatherogenic effect of VLDL- and IDL-apo E. The balance between these effects could be influenced by apo E phenotype. PMID- 9853026 TI - [Is microalbuminuria, an early marker of clinical nephropathy, also a cardiovascular risk factor?]. AB - Microalbuminuria is not only a predictor of diabetic nephropathy in type 1 diabetes, but also a potent marker of cardiovascular risk, especially in type 2 diabetes. Microalbuminuria also predicts cardiovascular morbidity in the general population. We describe semi-quantitative and quantitative methods for determination of low urinary excretion of albumin. Pathogenetic hypotheses common to both renal and endothelial dysfunction are discussed, suggesting that microalbuminuria may be a link between micro- and macroangiopathy. Improved glycemic control and antihypertensive treatment postpone and potentially prevent development of nephropathy in diabetic patients with microalbuminuria. These interventions must be instituted early in the development of diabetic nephropathy. In type 2 diabetes, prospective studies are needed to evaluate the precise impact of such a therapy on the cardiovascular risk. PMID- 9853025 TI - [Focalized matrix proteolysis and inflammation]. AB - The zinc metalloproteinases (MMPs or matrixins) are capable of damaging most of the constituents of the extra-cellular matrix and the basement membrane. The matrix proteolysis is the result of an imbalance both in the turnover of these constituants and in the ratio of the tissue inhibitors of metalloproteinases (TIMPs) versus metalloproteinases. After a brief description of the nature and structure of MMPs and TIMPs, this article reports on recent progress concerning the intra and extra-cellular activation mechanisms of proenzymes (proMMPs) which bring into play a series of proteolytic activations involving different proteinase families. Two points are stressed: 1) the main sites of focalized matrix proteolysis regulation, illustrated in the cellular interaction of inflammation, and 2) the wide phenotypic variety of MMPs and TIMPs. PMID- 9853027 TI - [Bartonellosis: I. Bartonella henselae]. AB - The recent discovery of the bacterium Bartonella henselae was mainly due to the development of molecular biology techniques adapted to microbial diagnosis and to the description of new human diseases linked to Aids. About 10% of pet cats and 33% of stray cats harbour that bacterium in their blood. In immunocompetent patients, that bacterium is responsible for human cat scratch disease, characterized essentially by a localized lymph nodes enlargement in the vicinity of the entry site of the bacteria. This disease occurs more likely in pet cats less than 1-year-old and infested with fleas. The bacterium is transmitted to humans by scratches or bites; the role of fleas is possible, but is not yet documented. In 5 to 13% of cases, the cat scratch disease appears as more severe, including health impairment, hepatitis, Parinaud's oculo-glandular syndrome, neurological complications or stellate retinitis. In immunocompromised patients, B. henselae is responsible for various clinical presentations: bacillary angiomatosis, bacillary peliosis, recurrent or persistent bacteremia or endocarditis. Diagnosis of infections due to B. henselae can be performed by serological specific testing with sensitivity and specificity values ranging from 75 to 100%. Cultivation of the bacterium is fastidious, particularly in cases of cat scratch disease. The most efficient diagnostic test is the in vitro DNA amplification which has the drawback to require a lymph node sample. Antibiotics are usually inefficient for the treatment of cat scratch disease. By contrast, in immunocompromised patients, these infections are successfully treated for a more or less long time by macrolides or tetracyclines or rifampin. PMID- 9853028 TI - [Virus resistance in a hospital environment: overview of the virucide activity of disinfectants used in liquid form]. AB - Human pathogenic viruses can be detected in the hospital environment, on contaminated surfaces or medical instruments. Their transmission to patients or staff has already been reported. Lipophilic viruses (HIV, HBV, HCV,...) are susceptible to many liquid chemicals, but they can survive during short time on inadequately disinfected surfaces. Hydrophilic viruses, without envelope, are more resistant, but generally not associated with severe illnesses. Viruses survival in environment depends on many factors and is always improved with viral aggregation and low temperature, whereas organic matters and relative humidity effects are contrasted. The mechanism of virucide disinfectants is not yet well established, and their targets are not known with precision. Different disinfection procedures (disinfectant concentration, contact time, temperature, pH) can provide a similar virucidal activity on a given virus. The virucidal activity of a disinfectant is evaluated with a cell culture assay in Afnor guidelines. But, there are three major problems with this method, concerning need of high viruses titers, residual disinfectant cytotoxicity on cell culture, and non cultivable viruses. Non standardized tests are also described in papers, but their results can generally not be compared. Molecular biology improvements may lead to reproducible and sensitive tests. At present, no general disinfection procedure effective for most of the viruses, without risks for staff or materials, and with an acceptable economic cost can be recommended. PMID- 9853029 TI - [Cardiovascular risk factors in ethiopian medical students]. AB - Selected cardiovascular risk factors, serum cholesterol and triglyceride levels, presence of overweight, blood pressure and lifestyle patterns were assessed in 66 male medical students from Ethiopia aged between 18 and 26 years. Mean serum cholesterol concentration was 134.4 mg/dl with a standard deviation (SD) of 39.2 mg/dl; mean triglyceride concentration was 93.8 mg/dl (SD 43.6 mg/dl) and mean HDL-cholesterol in a subsample (n = 26) was 41.4 mg/dl (SD 14.2 mg/dl). Eight students (12.1%) were classified as having a sedentary life-style, 1 student was hypertensive, 5 (7.6%) had a body mass index above 28 kg m-2, 6 (9.1%) had serum cholesterol levels above 200 mg/dl and 5 (7.6%) triglyceride levels above 180 mg/dl. There was a trend for some risk factors to increase with the socio economic status. In multiple regression analysis, body mass index was the only independent factor associated with systolic blood pressure, serum cholesterol and triglyceride concentrations, while body mass index itself was significantly related to parental income. PMID- 9853030 TI - [Assay of cardiac troponin T on Elecsys 2010: comparison with cardiac troponin I]. AB - Cardiac troponin T (troponin Tc) of second generation has been measured on the Boehringer Elecsys 2010 analyzer. Cardiac specificity has been studied in patients presenting a rhabdomyolysis syndrome and the results compared with those obtained for cardiac troponin I (troponin Ic) measured on the Dade-Behring Stratus analyzer. The results clearly demonstrated that both troponin Tc and Ic showed similar cardiac specificity. Moreover, troponin Tc and troponin Ic can be indifferently used for the biological diagnostic of myocardial infarction or to asses reperfusion after percutaneous transluminal coronary angioplasty for acute myocardial infarction. In renal disease, troponin Tc was upper the reference limit (0.10 microgram/l) in 25% of the patients studied (20 patients). By contrast, troponin Ic was upper the reference limit (0.6 microgram/l) in only one patient. PMID- 9853031 TI - [Value of PCR for the early diagnosis of atypical zoster]. PMID- 9853032 TI - [Determination of total proteinuria: a comparative study of automated trichloracetic acid and pyrogallol red techniques for samples with monoclonal light chains]. PMID- 9853033 TI - [A particular myelodysplasia: 17p- syndrome]. PMID- 9853034 TI - [When immunology helps cytology....]. PMID- 9853035 TI - [Macrophage activation in a 74-year-old man]. PMID- 9853036 TI - [Evaluation of high performance liquid chromatography modular system for the diagnosis of hemoglobinopathies]. PMID- 9853037 TI - [Heparin induced thrombocytopenia associated with thrombosis]. PMID- 9853038 TI - [GBEA service conselors]. PMID- 9853039 TI - [Obsolete emergency biology. The confluence of critical care analysis and near patient testing]. PMID- 9853040 TI - [Resistance to penicillin G and Streptococcus pneumoniae infection at the Hopital Foch, Paris, France, in 1995]. AB - Decreased susceptibility to penicillin G of pneumococcal strains is continuously increasing in France. OBJECTIVE: We assessed effect of resistance to penicillin on therapeutic management and mortality in adults with pneumococcal pneumonia in our hospital. METHODS: This one-year retrospective study (1995) included patients with proven pneumococcal infection (positive blood culture, pleural fluid, or specimens from the lower respiratory tract). Strains of Streptococcus pneumoniae were screened for susceptibility to antimicrobial agents. Resistance to penicillin G was defined as a minimal inhibitory concentration > or = 0.12 microgram/ml. Age immune and nosocomial status, first and second line antibiotherapy and death were compared according to the strains susceptibility to penicillin G. A p value below 0.05 was statistically significant. RESULTS: In 15 cases a pneumococcal strain susceptible to penicillin G was isolated while 23 patients were infected with a strain with a decreased susceptibility to penicillin G. Age was significantly higher in the latest group (61.6 versus 54.7 years) while no difference was noted between the 2 groups according to immune and nosocomial status, therapeutic management and death. DISCUSSION: Resistance to penicillin did neither appear to increase mortality nor to influence therapeutic management in patients with pneumococcal infection. PMID- 9853041 TI - [Emphysematous pyelonephritis]. AB - Pyelonephritis is a frequent infection of variable severity. Mortality is low since the era of antibiotics. Emphysematous pyelonephritis is however a rare but life threatening form of kidney infection, which affect more frequently diabetic subjects, characterized by gas producing bacteria. These gaseous lesions can be localized in the renal parenchyma or in the perirenal or retroperitoneal space. Diagnosis became easier with the advent of CT scan. Treatment usually requires surgery and antibiotics. We report a case of emphysematous pyelonephritis in a diabetic patient. The CT scan strongly suggested the diagnosis which was confirmed per-operatively. Proteus mirabilis is exceptionally described in emphysematous pyelonephritis. PMID- 9853042 TI - [Vaccinations]. PMID- 9853043 TI - [Vaccinations of the traveller]. AB - Travelers' immunization has 2 aims: for the traveler, to prevent the risk of contracting an endemic disease during his stay abroad; for the community to prevent the risk of importing an infectious agent yet unknown in the country. Travelling offers an opportunity to update routine immunizations: tetanus, diphtheria, poliomyelitis, hepatitis B; for young people: measles and rubella; for elderly people: influenza. Two vaccinations are compulsory: yellow fever for travelers to tropical Africa and Amazonian forest; meningococcus A + C for Mecca pilgrims. Other vaccines are recommended for travelers to specific areas: typhoid fever, hepatitis A, cholera in countries with poor hygiene; rabies for exposed travelers (expatriates, trekkers...); Japanese encephalitis for persons spending a month or longer in rural agricultural areas during the monsoon season; tickborne encephalitis for persons visiting forested areas of central Europe from may to september. Yet, most of travelers' diseases such as malaria cannot be prevented by vaccination and appropriate preventive measures (chemoprophylaxis and protection against insects) should be taken. PMID- 9853044 TI - [Vaccination against diarrheal diseases and typhoid fever. Current status and prospects]. AB - Diarrheal diseases and typhoid fever are still common in developing countries and there is still a search for effective control measures able to prevent the epidemics they cause from time to time. There are recommended preventive measures based on health education and improvement of sewage and water facilities; however these recommendations given for many years have not reached the expected results for different reasons. Antibiotherapy was very effective for many years; unfortunately increasing antibiotic resistance has been reported, particularly in Shigella and typhoid fever treatment. This explains the re-kindled interest currently taking place in vaccines development against infections due to V. cholerae, Shigella, E.coli ETEC, S. typhi and rotavirus. The new available vaccines are very effective and provide greater protection than that given by the old killed injectable vaccines. They paved the way for development of new candidate vaccines easier to deliver (oral vaccines or one dose parenteral vaccine), which already give promising results. Some of these candidate vaccines like those related to Shigella infections are considered as a future promising tool for controlling diarrhea due to Shigella. PMID- 9853045 TI - [Vaccinations and systemic diseases. Vaccinations and immunosuppression]. AB - The hypothesis of systemic disease flare up or onset triggered by vaccination cannot be confirmed by the analysis of data from the literature. In immune deficiency of any cause, immunization is of great interest to avoid preventable infectious diseases but the risk of adverse events and suboptimal immunologic responses has to be considered. For live, attenuated vaccines the main risks are postimmunization complications, specially in persons severely immunocompromised. For killed or inactivated vaccines there are no contraindications, but the immune response is reduced leading to consider higher vaccine doses or more frequent boosters. However the response to such modifications of vaccine schedules has not been systematically evaluated and firm recommendations cannot be made at this time. In addition, in HIV infection, the effect of vaccination on viral replication has to be considered. In developing countries some live vaccines are still recommended in HIV+ children because of the prevalence of the target diseases (measles, poliomyelitis, tuberculosis...) and the rarity of complications. PMID- 9853046 TI - [Risks associated with vaccinations]. AB - Since Jenner and Pasteur, various vaccines have been developed and administered in immunization program conducted by WHO in order to diminish the circulation of pathogenic agents and eradicate some diseases. Risks associated with immunization are revealed by the collection and assessment of adverse events reported after the use of these drugs. They vary according to the type of vaccines. With high rates of immunization and a low incidence of infectious diseases, adverse events receive increasing attention. Frequent and mostly expected adverse events are reported in clinical trials. Unexpected rare adverse events are reported after marketing authorization by spontaneous reporting and post marketing surveillance studies. Post marketing surveillance should be adapted to vaccines (vaccino vigilance) and should take into account the risk linked to the disease they may protect against. Adverse events are often temporally associated with vaccines, that does not mean they are causally related. Specific studies should be conducted to assess the causal relationship between vaccines and post immunization adverse events. In order to reduce the risk associated with immunization, a strict follow-up of recommendations, warnings and contraindications in addition to appropriate information being delivered to both vaccinees and physicians are required. PMID- 9853047 TI - [Expanded program of vaccination. Results and prospects]. AB - The expanded program on immunization, jointly launched by WHO and UNICEF in 1974 aimed in the beginning at immunizing 80% of the children of the world against measles, tetanus, pertussis, poliomyelitis, diphtheria and tuberculosis. After reaching the objectives in 1990, countries have been urged towards eradication of poliomyelitis, elimination of neonatal tetanus, and measles control. Immunization against hepatitis B and yellow fever were also proposed according to local epidemiology. Tremendous results have been already delivered: the American continent has eliminated poliomyelitis since August 1991, it has reduced dramatically the incidence of measles. All the countries, including those from Africa, are organizing campaigns aiming at polio eradication. Active surveillance systems have been implemented. But financial and operational constraints persist, linked to vaccines prices, their heat sensitivity, injection techniques, and to the sterilisation and waste disposal. Major efforts are still needed. PMID- 9853048 TI - [Vaccination calendar and new risks of infection]. AB - The French program for immunization has undergone great changes since the last 90', with new vaccines and strategy improvement. The resurgence of pertussis in young adults with transmission to young infants not adequately immunized is due to the lack of immunity and reduced potential to acquire exposure-induced immunity. A late booster is recommended at 11 years by the new acellular pertussis vaccine (combined with DT I.P.V. Vaccines). The coverage rate for measles-mumps-rubella vaccine has reached 82% for 2 year old infants, but the stability for many years has left many susceptible children (not immunized or not seroconverted). A recent mathematical study shows a next risk for large epidemics in older children, adolescents and adults. Rubella has the same development with a recent rise of rubella in pregnant women. The recent recommendation of Health authorities of giving a second dose of measles-mumps-rubella vaccine at 4-6 years is associated to necessary enhancement coverage in 2 year-old infants to 95%. Universal immunization against hepatitis B has been promoted in 1994 for infants and adolescents. The decision is related to the growing number of new hepatitis B infections in spite of a well conducted risk-subject immunization. The 1997 coverage is better for adolescents (78%) than for infants (36%). PMID- 9853049 TI - [Hepatic tuberculosis: a case with pseudotumours form]. PMID- 9853050 TI - [Spontaneous Escherichia coli infection of pleural effusion in alcoholic cirrhosis without ascites]. PMID- 9853051 TI - [Meningoencephalitis caused by Toxocara canis]. PMID- 9853052 TI - [What role do physical activity and sedentary life style play in development and maintenance of excess pounds in the child?]. PMID- 9853053 TI - [Could management of the pediatric victim of severe head injury be guided by research in the adult?]. PMID- 9853054 TI - [Molecular bases of glycogenoses]. PMID- 9853055 TI - [Fanconi disease: study of 43 cases in southern Tunisia]. AB - BACKGROUND: To report the epidemiologic, clinical, biological features and course of Fanconi's anemia in southern Tunisia. PATIENTS AND METHODS: During a period of 12 years we observed 43 cases. For each patient, careful clinical, biological (hemogram, myelogram, bone marrow biopsy, hemoglobin electrophoresis, karyotype) and radiological (skeleton X-rays, abdominal echography and intravenous urography) examinations were performed. All the patients who were at a pancytopenia stage were given androgens. None had a bone marrow allograft. RESULTS: There were 24 girls and 19 boys. The mean age at diagnosis was 10 years and 9 months. The familial character was present in 53% of the cases. The most frequent initial complaint was anemic syndrome (69%). In ten cases (24%), the diagnosis has been established during a familial investigation. Malformations were present in all cases (abnormal pigmentation: 86%; skeletal maturation retardation: 83%; facial dysmorphy: 76%; statural hypotrophy: 65%; bone abnormalities: 53%; renal malformations: 44%). Anemia was present in 88% of the cases, thrombocytopenia and neutropenia in all cases. Bone marrow was hypoplastic or aplastic in all cases on biopsies. Spontaneous chromosomal breaks were found in 79% of the studied cases. Fetal hemoglobin was increased in 80% of the studied cases with a mean level of 20.5%. Actuarial survival rate at 5 years was 48%, but long survival durations were rare (eight out of 43 patients). DISCUSSION: This disease, rare in the world, seems to be frequent in southern Tunisia. A normal karyotype (with classical techniques), found in five patients, could not discard the diagnosis; for this reason, the use of sensitizing agents should improve the sensitivity of the test. Besides, an increased level of fetal hemoglobin enabled us to suggest the diagnosis in some cases. Androgenotherapy increased the survival duration to more than 5 years in eight patients. However, bone marrow allograft remains the only possibility of cure. PMID- 9853056 TI - [Fetomaternal hemorrhage: a series of 9 cases]. AB - AIM: This study was designed to stress the importance of early diagnosis of fetomaternal hemorrhage (FMH) in attempt to prevent the subsequent adverse outcome on the fetus and the newborn. PATIENTS AND METHODS: Nine newborns were admitted because of neonatal anemia to our neonatal unit from October 1989 through September 1995. The diagnosis of FMH was made by the sigma diagnostic fetal hemoglobin that is the Kleihauer test in our hematologic laboratory. Other causes of neonatal anemia have been ruled out. RESULTS: Seven out of the nine cases have expressed early signs of fetal distress in term of abnormal fetal monitoring and/or thick meconium associated with decreased fetal movements. At birth, a wide clinical spectrum depending on the amount of the hemorrhage was seen, ranging from mild anemia with no symptoms (four cases), hypovolemic shock (one case), respiratory distress syndrome (two cases) and maladjustment to extra uterine life (one case). There was one death at 48 hours after birth; one infant survived with severe encephalopathy. CONCLUSION: These results indicate that it is mandatory to carry out a Kleihauer test whenever a high suspicious index of FMH is faced or an unexplained neonatal anemia is found. PMID- 9853057 TI - [Vitamin D status in the adolescent: seasonal variations and effects of winter supplementation with vitamin D3]. AB - BACKGROUND: Recently, 25 hydroxyvitamin D (25 OHD) blood concentrations measured in adolescents during or at the end of winter were found very low. A concomitant stimulation of parathyroid function was observed. The aim of the present study was to test the biological effects of a treatment with vitamin D3 during winter. POPULATIONS AND METHODS: The effects of vitamin D3 supplementation (100,000 IU, twice, at the end of November and of January) were assessed in 24 male Caucasian adolescents (mean age +/- SD: 14 y 6 m +/- 9 m). They were pupils in a lad jockeys training center located in the countryside near Chantilly (49 degrees northern latitude). Blood concentrations of 25 OHD, calcium and intact parathormone (PTH) were measured three times: before each oral intake of vitamin D3 and 2 months after the last intake (March). A group of 32 male adolescents (mean age +/- SD: 14 y 9 m +/- 6 m), pupils in the same center, receiving no vitamin D and sampled in November and in March, served as controls. RESULTS: In March, mean concentrations of 25 OHD (8.36 +/- 2.73 micrograms/L) were very low in vitamin D-not supplemented adolescents since 34% had levels less than 6 micrograms/L. In March, PTH concentrations (40.5 +/- 12.2 ng/L) were significantly (P = 0.0001) higher than in November (28.8 +/- 9.9 ng/L). In boys receiving vitamin D3 25 OHD serum concentrations measured in January (17.5 +/- 3.2 micrograms/L) and in March (18.7 +/- 4.0 micrograms/L) remained at a level not very different from that measured in November (16.6 +/- 3.8 micrograms/L). During the same period, calcium and PTH concentrations (32.2 +/- 11.7 ng/L in November; 32.4 +/- 14.3 in January and 32.9 +/- 13.5 ng/L in March) remained at their basal level as well. CONCLUSIONS: The observation that, after winter, a relatively large number of adolescents presented low concentrations of 25 OHD suggests that, during winter, usual dietary intakes and/or vitamin D stores are not sufficient to provide for their needs. Administration of two oral doses of 100,000 IU of vitamin D3 could maintain the vitamin D status at its initial level. The efficiency of such a prophylactic treatment is also assessed by its effect on parathyroid function. PMID- 9853058 TI - [Intensive nocturnal hemodialysis, a choice for survival in renal failure without hope for a graft]. PMID- 9853059 TI - [Bacteriologic surveillance of nosocomial septicemia and bacteremia in a pediatric hospital]. AB - BACKGROUND: Nosocomial bloodstream infections in pediatrics are an important cause of morbidity and mortality. To identify pathogens causing nosocomial bloodstream infections, evaluate associated risk factors and take preventive measures, we conducted a prospective study from January 1995 to December 1995 at Saint-Vincent-de-Paul Hospital (Paris). PATIENTS AND RESULTS: All patients hospitalized more than 48 hours were included in the study. During this period, we recorded 21 bloodstream infections in 20 children. The incidence rate of nosocomial bloodstream infection was 1/1,000 admissions. Sixteen children were hospitalized in surgery, three in medical intensive care unit; the median day onset of infection was approximately 20 days. Recorded risk factors were: surgery, invasive procedures, central catheterization, bladder catheters, parenteral nutrition, device, endotracheal tube, antibiotic therapy before infection. The number of risk factors ranged from zero to six per patient. The most common isolated pathogens were in ten cases Gram positive cocci: five methicillin-sensible Staphylococcus aureus, four methicillin-resistant coagulase negative staphylococci and one Streptococcus milleri. Other bacteria were seven enterobacteria, three Pseudomonas sp and three Candida sp. In 11 cases, the same bacteria as in bloodstream infection could be found: in three urine samples, in two tracheal samples, in two gastro-intestinal samples, two puncture sites, one device, and one umbilical catheter. CONCLUSION: In our study, 6.2% of positive blood culture were due to a nosocomial infection. We confirm the importance of Gram positive cocci, and particularly of methicillin-resistant coagulase negative staphylococci. PMID- 9853060 TI - [Risk of phosphate enemas in the infant]. AB - BACKGROUND: Hypertonic sodium phosphate enema (Fleet) are available for relief of constipation. They may be responsible for life-threatening electrolyte disturbances (hyperphosphatemia, hypernatremia, hypocalcemia) and severe dehydration. OBSERVATION: A 14-month-old child with a neonatal repaired Hirschsprung's disease was urgently admitted for apathy. The clinical diagnosis was sepsis from intestinal origin. A few hours before admission, the child had received a pediatric phosphate enema ("Fleet Enema"). The clinical symptoms and the observed electrolyte disturbances were the consequences of the intoxication by the enema. CONCLUSION: In children with renal insufficiency or bowel dysfunction, phosphate enemas are dangerous. Even in normal children, they should not be used under 2 years of age and only with extreme caution between 2 and 5 years. PMID- 9853061 TI - [Children who swallow air]. AB - Gas in the gut derives from swallowed air, intra-luminal production and diffusion from the blood. Excessive swallowed air may cause pathologic aerophagy. CASE REPORTS: This report describes four children aged from 3 to 12 years with pathologic aerophagy due to excessive air swallowing. One of them had a "Gas Bloat Syndrome". Clinical presentation was very incongruous. However, the clinical features were characteristic: abdominal distention increasing throughout the day, associated with visible and often audible air swallowing and excessive flatus. The questioning and clinical examination helped for diagnosing. Medical symptomatic treatment was associated with a psychotherapy approach. CONCLUSION: A better recognition of this condition might contribute towards earlier diagnosis. PMID- 9853062 TI - [Diagnosis of phenylketonuria in a 35-year-old mother in relation to prenatal diagnosis of intrauterine growth retardation with microcephaly]. AB - BACKGROUND: Some French pregnant women with phenylketonuria (PKU), born before 1978, have not been tested with the Guthrie method during the neonatal period. They have a risk of spontaneous abortion and their infants are often mentally retarded with microcephaly and/or congenital heart anomaly. CASE REPORT: A woman with a moderate mental retardation became pregnant at the age of 31 years. Her newborn had a severe intrauterine growth retardation with microcephaly and developed mental retardation. This mother became pregnant again 4 years later. Repeated fetal ultrasonography showed progressive growth retardation. Maternal blood phenylalanine concentration was 18 mg/100 mL. Therapeutic abortion at 27 weeks of gestational age showed a fetus with several abnormalities, particularly cardiovascular and cerebral. CONCLUSION: It is still possible to meet women with unrecognized atypical PKU. Fetus or infant born with unexplained growth retardation and microcephaly requires search for maternal PKU. PMID- 9853063 TI - [Chronic meningococcemia: 3 cases in the immunocompetent child]. AB - Chronic meningococcemia is a part of extra meningeal manifestations of meningococcal disease. Its diagnosis can be difficult because of lack of sensitivity of blood cultures. CASE REPORT: Three cases, concerning immunocompetent children, respectively aged of 14, 10 and 4 years are reported. The clinical course was characterized by recurrent fever, inflammatory joint manifestations and diffuse maculopapules secondary centered by petechiae. Microbiological findings revealed in one case a positive throat culture and presence of meningococcal soluble antigens in blood and urine. In the other two cases, diagnosis was done after done after positive blood culture at the 7th, and 13th days of course. CONCLUSION: The diagnosis should be considered in any children with a prolonged, recurrent fever and cutaneous and joint manifestations even if blood cultures remain negative. The response to therapy by usual antimeningococcal antibiotics is dramatic and curative while a prolonged untreated course may be complicated by metastatic infection. PMID- 9853064 TI - [Fetal echocardiography]. AB - Cardiac malformations are the most frequent and serious fetal malformations. However their prenatal diagnosis is the less easy. Fetal echocardiography is the main diagnostic tool; in addition it is helpful for the therapeutic orientation. Only 30% of fetal heart diseases are detected on account of maternal or fetal indications, most of them being discovered without any known risk factor. In order to improve its diagnostic sensitivity, fetal echocardiography must include a four chambers view and a study of ventricular and arterial connection. Nevertheless, many heart diseases remain undetected, most of them being minor with favorable prognosis. Search of an extracardiac abnormality and karyotype analysis must be performed in order to explicit the prognosis. Fetal arrhythmias can be analyzed by study of the atrial and ventricular contraction using M-mode echocardiogram or Doppler mode; most of them are supraventricular and accessible to maternal therapy. The sustained fetal bradycardia is frequently a complete heart block whose prognosis depends upon its association with a heart defect and/or a ventricular escape. Antibodies SS-A and SS-B are frequently associated with complete heart block without structural heart defects. Hypertrophic cardiomyopathy may be observed in fetus of diabetic mothers needing an accurate evaluation of the thickness of the interventricular septum. Echocardiography is a part of fetal medicine, which includes the different specialties devoted to the fetus. PMID- 9853065 TI - [Mucoviscidosis: therapeutic strategies are multiplying]. AB - Since the cloning of the defective gene in cystic fibrosis, much has been learned on the function of CFTR and on the mechanisms regulating its expression. Based on the current understanding of the processes involved in lung disease progression, a number of approaches have been developed using gene therapy and pharmacological agents. Several of these agents have been reported to restitute a function to CFTR with specific mutations. Other molecules act on channels other than CFTR, and may be effective by bypassing CFTR itself. In the present review the various therapeutical strategies currently investigated are discussed. PMID- 9853066 TI - [Radiology case of the month. Pulmonary hypertrophic osteoarthropathy in the course of Hodgkin's disease]. PMID- 9853067 TI - [What is new in pediatric pneumology?]. AB - Have been selected for this review: 1) the recent and impressive development of high-resolution and spiral CT scan in pediatric thoracic imaging; 2) the emerging of new and promising therapies for asthma (long acting inhaled beta 2-agonists, leukotriene antagonists, anti-IgE monoclonal antibodies); 3) the multifactorial origin of asthma in childhood; 4) the development of thoracoscopic surgery, a minimal-invasive approach beneficial in numerous circumstances. PMID- 9853068 TI - [Management of Rhesus isoimmunization. Viewpoint of the obstetrician]. AB - Although rare, severe forms of Rhesus isoimmunization are still observed. Early diagnosis and treatment with intrauterine transfusions allow an 80% survival rate. Anti-D alloimmunizations usually result from missed prophylaxis with anti-D serum at delivery, thus underlying the need for a rigorous application of this prophylaxis. PMID- 9853069 TI - [Pharmacologic treatment of neonatal jaundice. A new approach]. AB - Pharmacological treatment of neonatal jaundice is again topical. At the beginning of the eighties, clofibrate was added to phenobarbital which was difficult to use and inefficient. Clofibrate is a better enhancer of glucuronosyl transferase induction than phenobarbital and causes 100% increase of hepatic bilirubin clearance within 6 hours. In the treatment of early jaundice in full term neonate, it significantly reduces bilirubinemia in 16 hours, and decreases the intensity and duration of jaundice and also phototherapy requirement. At the end of the eighties, new molecules inhibiting hepatic production of heme to bilirubin, like metalloporphyrins, were introduced. These molecules block the transformation of heme to biliverdin and bilirubin. Among them, the Sn mesoporphyrin seems to have the best efficacy when used prophylactically in premature infants between 30 and 36 weeks of gestational age, and also curatively in full-term neonates, with minimal side effects. However the product is not yet manufactured and can not be used in pediatrics practice. Therefore clofibrate represents the only pharmacological treatment of neonatal jaundice actually available. PMID- 9853070 TI - [Pediatrics on the internet]. AB - It is easy to use Internet tools with a computer's mouse but it is more difficult to obtain available addresses in order to reply to concrete and accurate search. The aim of this article is to help new Internet users in their quest for pediatric informations and medical knowledge. PMID- 9853072 TI - [Cerebrocostomandibular syndrome]. PMID- 9853073 TI - [Teratogenicity of carbimazole]. PMID- 9853074 TI - [Morbidity and mortality in pediatric intensive care at the Brazzaville University Hospital Center after 5 months of war, October 1997-April 1998]. PMID- 9853075 TI - [Hypopituitarism during the first year of life. A collaborative Italian study]. AB - Literature data on statural growth effect of growth hormone (GH) treatment of children with GH deficiency (GHD) appears disappointing when treatment had late onset. It seems therefore that GH treatment should be started as early as possible, thus requiring early diagnosis. This study was undertaken: 1) to define the main characteristics of early onset GHD which may help the clinician for early diagnosis; 2) to evaluate the effect of a GH treatment on the statural growth of early onset GHD, when treatment is started before 1 year of age. Sixteen children with GHD diagnosed before 1 year of age were included in the study; all of them received a GH treatment before 1 year of age and for a minimal duration of 5 years; four had isolated GHD secondary to a GH gene deletion (type 1A GHD). Based on this series, the main anamnestic and clinical characteristics of early onset GHD are: frequent perinatal pathology, short birth height in more than half of the cases, relatively elevated birth weight in all cases, poor statural growth with neonatal onset, associated signs (hypoglycemia, special facies, micropenis, cryptorchidism, liver dysfunction). The effect of GH treatment (> or = 5 years) on statural growth was limited and transient in children with type 1A GHD, and resulted in an incomplete catch-up growth in children with sporadic GHD, their mean height remaining significantly below their target height. PMID- 9853076 TI - [Catch-up growth in growth hormone deficit children treated with GH since the first year of life]. AB - Long-term effects of growth hormone (GH) treatment were studied in 59 GH deficient (GHD) children, whose treatment was started before the age of 1 year, during the period 1978-1992, under the supervision of the France-Hypophyse Association. Mean duration of treatment was 8 +/- 3.6 years. At the last evaluation, 20 patients (34%) had a height above normal mean for age, and 50 (85%) had a height above the second lower standard deviation (SD) for age. Final height prediction performed in 29 patients was greater than target size (-0.35 +/ 1.4 SD vs -0.56 +/- 0.79 SD). These results show that early treatment of GHD infants allows normal statural growth in the majority of the cases; in addition, they underline the importance of early diagnosis of GH deficiency. PMID- 9853077 TI - [Does the growth hormone pharmacokinetic profile help to predict the treatment response?]. AB - Pharmacokinetics of subcutaneously administered recombinant growth hormone (rGH) were studied in nine GH deficient children at the time of the first rGH injection at the beginning of treatment. Serum GH levels were determined by four different methods: immunofunctional assay, immunofluorometric assay and two bioassays on Nb2 cells. The results showed similar profiles whatever the type of assay with a concentration peak reached 2 to 6 hours after subcutaneous injection; however, large individual variations in peak amplitude were observed. They were related to individual variations in the growth velocity during the first year of rGH treatment. These variations are possibly related to individual differences in rGH degradation at the site of injection. Study of rGH absorption profiles appears useful in the evaluation of rGH treatment in GH deficient patients. PMID- 9853078 TI - [Growth hormone deficiency. Treatment with growth hormone and body composition]. AB - Increased fat mass, decreased lean mass, muscular mass and bone mineral density are characteristic of the body composition in GH deficiency, GH treatment reverses these abnormalities. Body composition was determined in 20 young adults with GHD diagnosed in childhood, whose GH treatment was stopped 1 year earlier. Reevaluation of GH secretion in these patients showed that 12 remained GH deficient (confirmed GHD) while eight recovered normal GH secretion (transient GHD). One year after stopping the GH treatment, patients with confirmed GHD showed an increased fat mass as compared with value at the end of the treatment; in addition a decreased bone mineral content was observed in the patients with low physical activity. There was no increased fat mass in transient GHD; however, these patients presented with low bone mineral content, as previously reported in adults with history of delayed growth and adolescence. PMID- 9853079 TI - [Genes and intra-uterine growth retardation]. AB - Genetic analysis of growth from birth to adulthood shows the existence of a strong genetic component in the variance of size at particular milestones in the growth process, such as height at take-off, at peak velocity and at adulthood. While it is well known that postnatal growth is strongly genetically determined, the importance of genes in normal variations of prenatal growth is less known. Genetic analysis of prenatal growth in human is not an easy problem and weighing the genetic and non-genetic component in intra-uterine growth retardation an almost impossible task. It is now well known that adults who had a low birthweight or who were thin at birth, with a low ponderal index, tend to be insulin resistant and have an increased risk of developing non-insulin-dependent diabetes mellitus later in life. According to the thrifty genotype hypothesis, genes predisposing to type 2 diabetes mellitus are very likely to have been survival genes for our ancestors, helping them to store energy during long periods of starvation. Both epidemiological surveys of adults born after prenatal exposure to exposure to famine and biochemical investigations of insulin resistance in low birthweight children show that the association between a low birthweight and an increased risk of developing type 2 diabetes mellitus later in life has a genetic basis. While low birthweight infants have a decreased survival probability in infancy, having a small baby may have been a selective advantage during long periods of starvation. This could explain why the same genetic variants cause low birthweight phenotype and insulin resistance predisposing to non-insulin-dependent diabetes. PMID- 9853080 TI - [Somatotropin function in term and premature newborns during the first month of life]. AB - Neonatal somatotropic function is characterized by a discrepancy between elevated growth hormone (GH) levels and low IGF I levels. This study aimed at explaining this discrepancy, particularly to examining if it could result from low GH bioactivity. Serum concentrations of bioactive GH (bio GH), GH measured by radioimmunoassay (riGH), GH binding protein (GHBP), IGF I and IGF binding proteins (IGFBP) were determined in 27 premature and term newborns during the first month of life. At day 4, riGH and bio GH concentrations were elevated in both premature and term newborns as compared with normal prepubertal children; GHBP and IGF I levels were low, with a positive correlation with gestational age (P < 0.001). There was a positive correlation between GHBP and IGF I levels. IGFBP-1 and IGFBP-2 levels were elevated and negatively correlated with gestational age (P < 0.005). IGFBP-3 levels were within the range of prepubertal children values and positively correlated with gestational age (P < 0.005). During the first month, riGH and bio GH levels decreased in all infants, while IGFI levels increased in premature infants only, and GHBP levels in term infants only. The elevated levels of bio GH during the first days of life appear to be related to the low levels of IGF I due to a reduced number or function of GH receptors. In premature infants the decrease in GH levels observed afterwards appears to be secondary to the increase in IGF I levels. In term infants, in the absence of increase in IGFI levels other(s) factor(s) seem(s) to be involved. PMID- 9853081 TI - [Growth hormone treatment of children with short stature secondary to intrauterine growth retardation]. AB - There is now good evidence that short-term growth hormone treatment improves growth of children with short stature secondary to intrauterine growth retardation. A 3-year treatment with dosages greater than 1.2 U/kg/week allows these children to reach normal height, without any currently known side effect. Long-term efficacy, i.e., improvement of final height, is probable, but depends on the possibility of reconducting the treatment in case of loss of the initial benefits. The possible effect of treatment on puberty is presently unknown and will need special attention when treated children reach pubertal age. PMID- 9853083 TI - [Critical evaluation of growth hormone treatment in children with chronic renal insufficiency]. AB - Growth failure remains a major complication of chronic renal insufficiency in children, which greatly affects their quality of life. Based upon the data published in the literature it appears that growth hormone therapy improves growth in these children, with little secondary effects. However, some uncertainties persist concerning the safety and efficacy of this therapy, so that careful evaluations must continue. PMID- 9853082 TI - [Physiopathology of the somatotropin axis in chronic renal insufficiency]. AB - Many factors contribute to the growth failure of chronic renal failure: water and electrolytes disturbances, hypertonicity, phosphate or calcium wasting, secondary hyperparathyroidism, anemia, hypertension, metabolic acidosis, and malnutrition. In addition, the pubertal growth spurt is usually stunted. Growth hormone (GH) resistance is observed with low GH binding protein (GHBP) level, and normal or low IGF I levels despite elevated GH level. Elevated IGFBP levels may contribute to a reduced IGF activity, especially in dialysed patients. Glucocorticoid therapy in transplanted patients further contribute to poor growth and inhibited IGF I activity. As conventional treatments have a limited effect to improve growth, adult height is often far below -2 SD. GH therapy has proved to be successful, especially in young children, overpassing the hormonal resistance so that an adult height within the normal range may be reached. PMID- 9853084 TI - [Growth hormone treatment of familial hypophosphatemic rickets]. AB - X-linked hypophosphatemic rickets (XLHR) is frequently associated with growth retardation and short adult stature, even with an appropriate conventional treatment associating phosphate and calcitriol or 1 alpha-hydroxyvitamin D. Its pathogenesis is unclear; growth hormone (GH) secretion is usually normal. Six children with XLHR and growth retardation were treated with GH during 6 years. In addition, they received the conventional treatment. At the beginning of the treatment mean age was 7.8 +/- 1.8 years, and height mean Z score was -3.4 +/- 0.5. A control group was composed of six children with XLHR (age: 7.9 +/- 2.5 years) receiving the conventional treatment only. Under GH treatment statural growth was improved, with significant increase in Z score and predicted adult height; the height gain was significantly higher in the GH treated group as compared with the group receiving the conventional treatment only. In addition, radial bone mineral density increased significantly under GH treatment. GH treatment thus appears to be a useful treatment to improve statural growth in children with XLHR. PMID- 9853085 TI - [Phenotype-genotype relations in growth hormone insensitivity]. AB - Growth hormone (GH) insensitivity is associated with several different mutations of the GH receptor gene and a recently described new genetic disorder of the IFGI gene. The phenotype and biochemical characteristics were studied in 82 patients with growth hormone insensitivity, from 23 different countries, with a mean age of 8.25 years. Mean height SDS was -6.09. SDS of the IGF binding protein -3 (IGF BP3) was 7.99. Twenty three per cent of the patients were GH binding protein (GHBP) positive (> 10%). Mean height SDS score was -6.5 in the GHBP negative patients and -4.9 in the GHBP positive patients (p < 0.001). Fifteen different mutations of the GH receptor gene were identified in 27 patients. There were no relationships between the type of mutation or the involved GH receptor gene exon and height or IGFBP-3 SDS. The new phenotype due to a partial deletion of the IGFI gene was described in a 15-year-old boy who presented with a severe intrauterine growth retardation, a very poor postnatal statural growth, a neurosensorial deafness and a mild mental retardation. He had elevated GH levels, normal levels of IGFBP3, undetectable levels of IGFI, and showed no response to GH treatment. A partial deletion concerning the exons 4 and 5 of the IGFI gene was found. Thus, GH insensitivity is associated with large variations in the clinical and biochemical phenotypes. PMID- 9853086 TI - [Effect of hormonal treatment on bone mineralization in Turner syndrome]. AB - A review of the literature together with personal results show that both growth hormone (GH) and estrogen treatments improve bone mineral density (BMD) in patients with Turner syndrome. Insofar as GH treatment alone appears to normalize BMD in girls with Turner syndrome it is suggested that substitutive estrogen treatment could be delayed in order to guarantee optimal result for their statural growth, without affecting the quality of their bone mineralization. PMID- 9853087 TI - [Activating mutations of the TSH receptor: a synthesis highlighting certain pediatric aspects]. AB - Mutations of the TSH receptor gene cause constitutive activation of the TSH receptor responsible for cases of familial hereditary hyperthyroidism (germline mutations), cases of sporadic congenital hyperthyroidism (de novo mutations), and thyroid hyperfunctioning autonomous adenomas (somatic mutations). The discovery of these mutations not only clarifies the pathogenesis of some forms of thyroid autonomy, such as cases of persistent neonatal hyperthyroidism without maternal thyroid autoimmunity, but also contributes to a better understanding of the structure-function relationships of the TSH receptor. PMID- 9853088 TI - [Activating mutations of the gonadotrophin receptors]. AB - The different types of activating mutations of LH and FSH receptors genes are described. They result in a constitutive permanent activation of the LH or FSH function responsible for functional disorders which is also observed in some ovarian tumours. Two types of functional disorders have been reported: male precocious puberty through activating mutation of the LH receptor, male fertility in the absence of FSH through activating mutation of the FSH receptor. Activating mutations of the FSH receptor observed in certain ovarian tumours result in hypersecretion of oestrogens. PMID- 9853089 TI - [Activating mutations of the parathyroid hormone and parathyroid hormone related peptide]. AB - Parathyroid hormone (PTH) and PTH related peptide (PTHrP) have a common main receptor: type I PTH-PTHrP receptor. PTH expresses its main metabolic actions, and PTHrP part of its autocrine or paracrine actions through this receptor. Jansen chondrodysplasia is a very rare disease mainly characterized by severe metaphyseal growth disorders leading to dwarfism and hypercalcemia. The group of Juppner from the Massachusetts General Hospital in Boston recently demonstrated mutations on the gene of the type I PTH-PTHrP receptor in five patients with Jansen chondrodysplasia. These mutations result in permanent activation of the receptor responsible for the observed hypercalcemia and bone growth abnormalities due to the disease. Therefore Jansen chondrodysplasia appears as a remarkable clinical model outlining the major role of PTHrP in bone growth regulation. PMID- 9853090 TI - [New growth hormone secretagogues: from physiology to the first therapeutic applications]. AB - Growth hormone releasing peptides (GHRP) are synthetic hexapeptides that physiologically stimulate GH release through two different pathways: 1) central and 2) direct action on somatotropic cells. Animal experiments and first clinical trials show that synthetic GHRP and synthetic analogues could be useful substitutes to recombinant GH in the treatment of GH deficiency, and in pathological conditions which may benefit from amplification of the GH-IGF I axis activity. PMID- 9853091 TI - [Genetic diseases in the Mediterranean region: a historical perspective]. PMID- 9853092 TI - Object-related attention: an event-related potential study. AB - In two experiments reaction time (RT) and event-related potentials (ERPs) were measured to visual stimuli consisting of two objects (Duncan, 1984; Vecera & Farah, 1994). Target stimuli were defined by two features, which either were distributed between the two objects or belonged to a single object. In Experiment 1 the two objects appeared in spatial overlap, whereas in Experiment 2 the two objects were spatially separated. In both experiments RT was shorter in the single-object condition. The late positive component of the ERP (P3) emerged, peaked, and declined earlier in the single-object condition. It is suggested that object-related attention may facilitate the earlier onset of search for the relevant features. PMID- 9853093 TI - Chronic electrical stimulation of the left ventrointermediate (Vim) thalamic nucleus for the treatment of pharmacotherapy-resistant Parkinson's disease: a differential impact on access to semantic and episodic memory? AB - Thalamotomy for medically refractory Parkinson's disease (PD) is considered to be efficacious and relatively safe. Because a minority of patients experience decrements in language and memory (often mild and transient) after thalamotomy, chronic thalamic deep brain stimulation (DBS) might be a safer treatment given its reversibility and the modifiability of stimulation parameters. Two preliminary studies support the relative cognitive safety of unilateral DBS of the ventral intermediate (Vim) thalamic nucleus, but it is unclear whether possibly subtle changes in language and memory represent effects of "microthalamotomy" or of stimulation per se. This report provides preliminary data concerning effects of left thalamic stimulation on information processing speed, semantic memory (verbal fluency and visual confrontation naming), and verbal episodic memory in a patient with PD. In addition to being evaluated before and 3 and 6 months after surgery, the patient was tested 18 months after surgery either on or off medications and with the stimulator turned either on or off (order counterbalanced across medication conditions). Test performance differences between the stimulation conditions were attenuated "off" as compared to "on" medication. Vim stimulation consistently, albeit subtly, improved semantic verbal fluency but interfered with immediate recall of word lists. Parallels to findings from acute, intraoperative thalamic stimulation studies are explored. The hypothesis is offered that left Vim stimulation might facilitate access to semantic memory, but interfere with episodic memory processes. PMID- 9853094 TI - Interpreting ambiguous advertisements: the effect of frontal lobe damage. AB - Despite intact primary language processes patients with frontal lobe deficits often have impaired communication skills including impaired capacity to understand conversational inference. This study examined the ability of three patients with demonstrated frontal lobe pathology to interpret lexically ambiguous advertisements. When compared to a nonbrain-damaged control group it was found that the frontal lobe patients were poorer at comprehending the abstract or inferred meanings inherent in the advertisements. The pattern of performance across the patients did, nevertheless, differ despite a similar end result. These findings are discussed in relation to theories concerning the contribution of the frontal lobes to language function. PMID- 9853095 TI - Comparative assessment of distance processing and hemispheric specialization in humans and baboons (Papio papio). AB - This comparative study explored the ability to process distance and its lateralization in humans and baboons. Using a conditional matching-to-sample procedure in a divided-field format, subjects had to decide whether or not the distance between a line and a dot belonged to a short- or a long-distance category. Experiments 1, 2, and 4 demonstrated the ability of baboons to process and categorize distances. Moreover, humans showed better distance processing for right visual field/left hemisphere presentations than for left visual field/right hemisphere (LVF-RH) displays (Experiments 1-2). The same bias was found in baboons (Experiment 1), but in a weaker way. In Experiment 3, naive human individuals were tested and the difficulty of the discrimination was enhanced. There was a LVF-RH advantage which vanished with practice. Results are discussed by referring to theories (i.e., Kosslyn, 1987) of visuospatial processing for coordinate and categorical judgments. PMID- 9853096 TI - Awareness of remembering achieved through automatic and conscious processes in multiple sclerosis. AB - Fifty multiple sclerosis (MS) and forty-one matched control subjects were administered a word stem completion task and the California Verbal Learning. Test (CVLT). As predicted, priming in a conventional "implicit" task did not differ for MS and control groups, and under "explicit" instructions verbal list recall was significantly lower in the MS group with recognition intact. Application of the process dissociation procedure (Jacoby, Toth, & Yonelinas, 1993) to the word stem task demonstrated that conscious and unconscious processes both contribute to remembering in MS. Exploratory analyses found no group differences in words remembered under inclusion nor exclusion conditions, though preliminary estimates for MS automatic processing exceeded those for controls. As predicted, prospective metamemory judgments reflected subsequent performance for both groups in the inclusion but not the exclusion condition. By contrast, retrospective monitoring for both groups reflected memory performance in both conditions. However post hoc analyses demonstrated that those MS subjects who employed more automatic processing were less aware retrospectively of their successful remembering under exclusion than MS subjects who used less automatic processing. PMID- 9853097 TI - The role of hemispheric activation in person perception: evidence for an attentional focus model. AB - Two studies assessed the role of hemispheric activation in person perception. Results of Study 1 supported an attentional focus model of hemispheric differences in person perception as opposed to a stereotyping/individuating model. Relative to left hemisphere-activated participants, right hemisphere activated participants made more individuating judgments and engaged in more trait level stereotyping, whereas left hemisphere-activated participants engaged in more global stereotyping. In Study 2, greater individuation by participants under higher levels of motivation and ability was shown to be mediated by relatively greater right hemisphere activation. Results of both studies are interpreted in terms of an attentional focus model of person perception. Implications for dual process models of social perception are also discussed. PMID- 9853098 TI - Ideomotor apraxia in early Alzheimer's disease: time and accuracy measures. AB - Twenty-six Alzheimer's disease (AD) and 42 healthy control (NC) subjects were evaluated with neuropsychological and apraxia batteries. ADs produced a greater range of error types, but did not differ from NCs in their most frequent error types. Hand sequencing ability contributed significantly to AD praxis with no predictors for NCs. Although groups did not differ in gesture time, the AD group had significantly longer response latencies for periods prior to gesture execution and the effect was prominent for transitive tasks and nondominant hand use. Results illustrate the sensitivity of timing measures in identifying abnormal praxis in early stages of AD. PMID- 9853099 TI - Fundamentals of process neuropsychology. AB - An examination of the whole-to-part transition over phases, from potential to actual in the specification of a concrete entity, as in the momentary mind/brain state, reveals patterns of change that can be considered a first approximation to the foundational laws of cognition. These laws, which amount to a theory of universal change, apply as well to the becoming or actualization of non-cognitive entities. Thus, the thesis is advanced that the mental and the physical actualize a generic process and that a theory of this process, process monism, is a metaphysics of the antecedents of occasions of fact or the laws of change that deliver objects. The commonality of the mental and the physical lies in the conceptuality of the duration of becoming and its continuity with the duration of the conscious present. The before/after relation that characterizes the phase transitions in a non-cognitive entity is the seed of the past/present relation in consciousness. The connectedness of past and present arises as a feeling of the relation of antecedent phases imminent in a concrete particular. The theory rejects as regressive the elimination of consciousness by a reduction to the material, or the reverse, in idealism, as well as an emergence of consciousness from material states. A deep current of connectedness runs from the nature of conscious phenomena to the categories of existence at the level of the atom. PMID- 9853100 TI - The role of speed of processing, inhibitory mechanisms, and presentation order in trail-making test performance. AB - The roles of speed of processing, inhibition, and presentation order in the Trail Making Test (TMT) performance were examined. One-hundred ten undergraduates performed the TMT (1/2 received order Part A-Part B, 1/2 received order Part B Part A) and also completed computerized tests of inhibitory functioning and speed of processing. Neither speed of processing nor inhibitory functioning affected TMT performance. However, order of presentation significantly affected TMT performance. Presentation order is an important variable to consider in TMT performance. PMID- 9853101 TI - A note on the possibility of explaining why a color cannot be both red and green. AB - To provide a neurophysiological basis for the opponent nature of color vision it has been previously argued that a color cannot be both red and green because color-opponent neurons cannot respond to both red and green at the same time. The present analysis shows that such arguments hinge on the possibility of excluding statements of the kind "a color can be both red and green." For an empirical fact to exclude such statements, these statements would have to be meaningful. However, statements like "a color is both red and green" are not meaningful and are not allowed in our language. Thus, the properties of neurons are not in a position to exclude the possibility of "a color that is both red and green." This means that this attempt to establish a neurophysiological basis for opponent colors is flawed. PMID- 9853102 TI - The differential effects of cueing on recall in Parkinson's disease and normal subjects. AB - Patients with Parkinson's disease (PD) are known to be impaired in supraspan verbal learning due to reduced availability of internal encoding capacity. PD patients may suffer from poor semantic, source and sequential processing capacity. Thirty-three patients with idiopathic PD and 42 matched normal control subjects (NC) were administered the California Verbal Learning Test under 3 conditions of graded cueing. While the PD patients benefited significantly and progressively from increasingly explicit cueing, the NC group did not since their performance was optimal even without cueing. Results are discussed in relation to frontal executive control of attentional resources and the establishment of encoding strategies. PMID- 9853103 TI - Line bisection and neglect severity. AB - The relationship between (horizontal) line length and bisection accuracy continues to be of theoretical interest in the study of visuospatial neglect. In a recent paper, Koyama, Ishiai, Seki and Nakayama (1997) claim that line bisection performance by patients with severe neglect is "not only quantitatively but also qualitatively different" from that of patients with moderate or mild neglect. In particular, they argue that line length does not control bisection displacements in severe neglect. Contrary to that position, I first demonstrate that the conclusion of Koyama et al. (1997) does not follow from their own data; second, I describe empirical results showing that bisection displacement in severe neglect is exquisitely sensitive to line length. The standard psychophysical models of line bisection in visuospatial neglect are not falsified by the results of Koyama et al. PMID- 9853104 TI - Collaterals of recurrent laryngeal nerve fibres innervate the thymus: a fluorescent tracer and HRP investigation of efferent vagal neurons in the rat brainstem. AB - The origin and course of efferent vagal fibers, which innervate the rat thymus, were investigated by a fluorescent retrograde double labeling method, using Fast blue (FB) and Diamidino yellow dihydrochloride (DY) as tracers. In the same animal, one tracer was injected into the cranial portion of the right lobe of the thymus and the other dye was deposited around the cut end of the right recurrent laryngeal nerve. The neuronal population giving origin to the recurrent nerve was mapped by using retrograde labeling with HRP applied to the central stump of the nerve. The HRP retrograde axonal transport showed that most efferent vagal fibers of the recurrent nerve have their perikarya in the nucleus retroambigualis (NRA), nucleus ambiguus (NA), and to a lesser extent in the nucleus retrofacialis (NRF). In fluorescent retrograde double labeling of thymus and recurrent laryngeal nerve both single and double labeled cells were found. The cells labeled by the injections into the thymus were colocalized with the neurons labeled by the tracer deposited in the recurrent laryngeal nerve to the NRA, NA, and NRF. Moreover along the rostrocaudal extent of the NRF and NA double labeled cells were present, showing that some of the thymic efferents are collaterals of the recurrent nerve fibers. Our experiments shown that some thymic vagal fibres originate from neurons of nucleus dorsalis nervi vagi (NDV) as demonstrated both by HRP and FB injected thymuses. The possible role of these efferents in thymic function is briefly discussed. PMID- 9853105 TI - Blockade of bepridil on IA and IK in acutely isolated hippocampal CA1 neurons. AB - The effects of bepridil, an antianginal agent with antiarrhythmic action, on voltage-dependent K+ currents in the CA1 pyramidal neurons acutely isolated from rat hippocampus were studied by means of whole-cell patch clamp techniques. Current recordings were made in the presence of TTX to block Na+ current. Depolarizing test pulses activated two components of outward K+ currents: a rapidly activating and inactivating component, IA; and a delayed component, IK. Results showed that bepridil reduced the amplitude of IA and IK, and exerted its inhibitory action in time- and dose-dependent manner. Half-blocking concentrations (IC50) of bepridil on IA and IK were 17.8 microM and 1.7 microM, respectively. 10 microM bepridil suppressed IA and IK by 46.7% and 77.1% at +30 mV of depolarization, respectively. When IK was activated nearly uncontaminated with IA by holding at -50 mV, 10 microM bepridil inhibited IK by 71.6% at +30 mV of depolarization; 10 microM bepridil positively shifted the voltage-dependent of activation curves of IA and IK 12.1 mV and 28.7 mV, respectively. These results suggested that blockade on K+ currents by bepridil is preferential for IK, and contributes to the protection brain against ischemic damage. PMID- 9853106 TI - Ca(2+)-dependent enhancement of [3H]noradrenaline uptake in PC12 cells through calmodulin-dependent kinases. AB - Ca(2+)-dependent regulation of [3H]noradrenaline ([3H]NA) uptake through the NA transporter was studied using PC12 cells. Preincubation for 10 min in the presence of 0.3-10 mM ca2+ in Krebs-Ringer (KR) buffer induced marked enhancement of the uptake (at 1 mM Ca2+, 6.6 times greater than that observed in the absence of Ca2+), which reflected both an increase in Vmax and a decrease in K(m) of the uptake process. Preincubation with 1 mM Ca2+ also induced a significant increase in the Bmax and Kd of [3H]desipramine binding. The uptake was still enhanced after washing cells with Ca(2+)-free buffer following preincubation with 1 mM Ca2+. 1-[N, O-bis(5-Isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62), 2-[N-(2-hydroxyethyl)-N-(4-methoxybenzenesulfonyl)]amino-N-(4-c hlo rocinnamyl) -N-methylbenzylamine (KN-93) (inhibitors of Ca2+/calmodulin-dependent kinase II), N-(6-aminohexyl)-5-chloro-1-naphthalenesulonamide (W-7) (a calmodulin antagonist), wortmannin (a myosin light chain kinase inhibitor) significantly reduced Ca(2+)-dependent enhancement of the uptake. Mycalolide B (an inhibitor of actin-myosin interaction) also inhibited the enhancement. Although calphostin C (a protein kinase C (PKC) inhibitor) did not affect the enhancement, 12-o tetradecanoylphorbol 13-acetate (TPA) inhibited the uptake. A synthetic peptide with a sequence (KKVIYKFFS579 IRGSLW) contained in the intracellular COOH terminal domain of a rat NA transporter was phosphorylated by purified brain Ca2+/calmodulin-dependent protein kinase II. These results suggest that Ca(2+) dependent enhancement of the [3H]NA uptake in PC12 cells are mediated by activation of calmodulin-dependent protein kinases, probably through stimulation of translocation of the NA transporter to the plasma membrane and/or direct phosphorylation of the transporter itself. PMID- 9853107 TI - Effect of bilateral lesions of the suprachiasmatic nucleus on hyperglycemia caused by 2-deoxy-D-glucose and vasoactive intestinal peptide in rats. AB - In mammals, the brain usually uses glucose as a sole energy source. Thus, under a central glucopenic condition after intracranial injection of 2-deoxy-D-glucose (2DG), an inhibitor of glucose utilization, it has been shown that rats elevate their blood glucose level through excitation of the sympathetic nerves. Experiments were conducted with rats to examine the role of the hypothalamic suprachiasmatic nucleus (SCN) in the hyperglycemic response to intracerebroventricular injection of either 2DG or vasoactive intestinal peptide (VIP). It was observed that, (1) intracerebroventricular injection of a VIP antagonist inhibited the hyperglycemic and hyperglucagonemic responses to the intracranial injection of 2DG; (2) bilateral electrolytic lesioning of the SCN suppressed the hyperglycemic and hyperglucagonemic responses to intracranial injection of 2DG, and intracerebroventricular injection of VIP restored these responses to 2DG; and (3) bilateral electrolytic lesioning of the SCN also suppressed the hyperglycemic and hyperglucagonemic responses to the VIP injection, and additional intracerebroventricular injection of 2DG caused hyperglycemia. These findings indicate that in rats with bilateral lesions of the SCN intracranial injection of 2DG is able to elicit hyperglycemia when VIP was administered intracranially, and suggest that neurons containing VIP-like immunoreactive substance (VIP-neurons) in the SCN have an important role in the mechanism of hyperglycemia elicitation following intracranial injection of 2DG. Moreover, these findings show that 2DG and VIP are able to realize their functions through acting on the brain sites outside the SCN. PMID- 9853109 TI - Effects of GABAB receptor antagonism on the development of pentylenetetrazol induced kindling in mice. AB - Pentylenetetrazol (PTZ) administered chronically in rodents induces kindling which is considered to be a model of chronic epilepsy mediated through a specific interaction with the GABA-gated chloride ionophore. PTZ kindling also impairs shuttle-box learning indicating a possible modulation of memory storage [A. Becker, G. Grecksch, H. Mathies. The influence of diazepam on learning processes impaired by pentylenetetrazol kindling. Naunyn-Schmiedeberg's Arch. Pharmacol. 349 (1994) 429-496]. Since GABAB receptor antagonism has been shown to improve cognitive performance in rodents and primates we have examined the effects of 3 antagonists; CGP 36742 (3-amino-propyl-n-butyl-phosphinic acid), CGP 56433 ([3-?1 (S)-[?3-(cyclohexylmethyl) hydroxyphosphinyl]-2-(S)-hydroxypropyl] amino]ethyl]benzoic acid) and CGP 61334 ([3 ?[3[(diethoxymethyl)hydroxyphosphinyl]-propyl-amino?meth yl]-benzoic acid) on the induction of PTZ kindling in mice at 48 h intervals for 8 weeks. Subsequently the mice were tested in an active avoidance paradigm. At the end of the experiment GABAB receptor autoradiography was performed on brain sections from these animals. Seizure intensity increased progressively in control mice reaching by 8 weeks a mean score which corresponded to clonic seizures. The GABAB antagonists suppressed kindling during the first 4 weeks and after that restored the seizure intensity to the level of control animals. The level of kindling was proportional to the avoidance score. The density of GABAB receptor binding in brain sections from PTZ kindled mice was significantly greater than in controls. This was not altered by pretreatment with the GABAB antagonists except in the cerebellum. PMID- 9853108 TI - Expression of glial cell line-derived neurotrophic factor and GDNFR-alpha mRNAs in human peripheral neuropathies. AB - The steady-state mRNA levels of glial cell line-derived neurotrophic factor (GDNF), GDNFR-alpha and RET were examined in various human peripheral neuropathies to determine the relationship with myelinated fiber pathology, and T cell and macrophage invasions in the diseased nerves. GDNF and GDNFR-alpha mRNA levels were elevated to variable extent in the diseased nerves, although they were not specific to the type of diseases. The increase of GDNFR-alpha mRNA levels was correlated with the extent of the nerves with axonal pathology, and was proportional to the extent of invasion of the nerves by T cells and macrophages. The GDNF mRNA levels were not related to axonal, demyelinating pathology, or inflammatory cell invasions. RET mRNA expression was not detected in normal nor diseased nerves. The GDNF and GDNFR-alpha expression in the diseased human nerves is regulated by an underlying pathology-related process, and could play a role in peripheral nerve repair. PMID- 9853110 TI - Prolonged seizure suppression by a single implantable polymeric-TRH microdisk preparation. AB - Thyrotropin-releasing hormone (TRH; Protirelin) is an endogenous neuropeptide known to have anticonvulsant effects in several seizure models and in intractable epileptic patients. Like most neuropeptides, its duration of action may be limited by a lack of sustained site-specific bioavailability. To attempt to provide long-term delivery, we attached TRH to a biodegradable polyanhydride copolymer as a sustained-release carrier. Utilizing the rat kindling model of temporal lobe epilepsy, a single TRH microdisk implanted stereotaxically into the seizure focus (amygdala) significantly suppressed kindling expression when assessed by the number of stimulations required to reach each behavioral stage and to become fully kindled (8.63 +/- 0.92 vs. 16.17 +/- 1.37; Mean +/- S.E.M.). Two indices of seizure severity, afterdischarge duration (Mean +/- S.E.M., sec.) (stimulated amygdala [87.40 +/- 5.47 vs. 51.80 +/- 15.65] and unstimulated amygdala [89.60 +/- 5.55 vs. 48.67 +/- 15.8] and clonus duration (71.2 +/- 5.94 vs. 29.40 +/- 8.87; Mean +/- S.E.M., sec.), were also significantly reduced by a single polymeric-TRH implant. Fifty days after initiation of the study a significant reduction in clonus duration (53.90 +/- 3.27 vs. 40.09 +/- 4.14) still remained in the TRH-implanted groups. This report is the first to provide evidence in support of in situ microdisk pharmacotherapy for potential neuropeptide delivery in intractable epilepsy and possibly other neurological disorders. PMID- 9853111 TI - The retinal projections to the ventral and dorsal divisions of the medial terminal nucleus and mesencephalic reticular formation in the Japanese monkey (Macaca fuscata): a reinvestigation with cholera toxin B subunit as an anterograde tracer. AB - After a monocular injection of the cholera toxin B subunit (CTB) into the vitreous chamber of the eye, retinal projections to the medial terminal nucleus (MTN) of the accessory optic system (AOS) were studied in the Japanese monkey. The anterogradely transported tracer was visualized with the peroxidase antibody technique by using an anti-cholera toxin antibody. One small accumulation of the CTB-immunopositive retinofugal terminals was located in a small area just medial to the medial edge of the cerebral peduncle and anterior to the attachment of the oculomotor nerve, suggesting the existence of a ventral division of the MTN of the AOS. Caudally, one very small bundle of the retinofugal fibers extending dorsally from this accumulation was seen running along the medial edge of the cerebral peduncle and substantia nigra to the small region corresponding to the dorsal division of the MTN. A few small bundles of CTB-immunopositive retinal fibers were observed to leave the superior fasciculus of the AOS at various points. These fibers coursed medially through the cerebral peduncle and substantia nigra to reach some restricted areas of the mesencephalic reticular formation between the medial lemniscus and the substantia nigra. PMID- 9853112 TI - The pre-Botzinger complex and phase-spanning neurons in the adult rat. AB - To characterise respiratory neurons in the pre-Botzinger complex of adult rats, extracellular recordings were made from 302 respiratory neurons in the ventral respiratory group of sodium pentobarbitone anaesthetised adult rats. Neurons were located 0 to 1.6 mm caudal to the facial nucleus, and ventral to the nucleus ambiguus. The pre-Botzinger complex comprised expiratory neurons (22%, 22/100), inspiratory neurons (37%, 37/100) and phase-spanning neurons (41%, 41/100). In contrast, 80% (125/157) of Botzinger neurons were expiratory, and 80% (36/45) of rostral ventral respiratory group neurons were inspiratory. Rostrocaudally, the pre-Botzinger complex extended about 400 microns, starting at the caudal pole of the nucleus ambiguus compact formation. The pre-Botzinger complex was also characterised by a predominance of propriobulbar neurons (81%, 13/16). Furthermore, 68% (33/48) of expiratory-inspiratory neurons found were located within the pre-Botzinger complex. The variety of neuronal subtypes in the pre Botzinger complex, including many firing during the expiratory-inspiratory transition is consistent with the hypothesis that this nucleus plays a key role in respiratory rhythm generation in the adult rat. PMID- 9853113 TI - Activation of metabotropic glutamate receptors is necessary for transneuronal regulation of ribosomes in chick auditory neurons. AB - Elimination of auditory nerve activity results in atrophy and death of nucleus magnocellularis (NM) neurons in the chick. One early event in the degeneration of NM neurons is a disruption of their ribosomes. This experiment examines the role of metabotropic glutamate receptors in afferent regulation of ribosomes. The auditory nerve on one side of a chick brainstem slice was stimulated in vitro. Rapid stimulation-dependent changes in ribosomes were visualized by immunolabeling using an antibody, called Y10B, that recognizes ribosomal RNA. In normal media, NM neurons on the stimulated side of the slice show greater Y10B labeling than the unstimulated NM neurons on the opposite side of the same slice. The role of metabotropic glutamate receptors was evaluated by unilaterally stimulating the auditory nerve in media containing the metabotropic glutamate receptor antagonist (RS)-alpha-methyl-4-carboxyphenyl-glycine (MCPG). Addition of MCPG to the bath did not block EPSPs produced by stimulating the auditory nerve. However, MCPG did prevent the stimulation-dependent regulation of ribosomes in NM neurons (as indexed by Y10B labeling). These data suggest that glutamate may play a trophic role in the young auditory system through activation of metabotropic glutamate receptors. PMID- 9853114 TI - NMDA receptor subunit NR1-immunoreactivity in the rat pons and brainstem and colocalization with Fos induced by nasal stimulation. AB - In the present study, we examined the distribution of neurons in the parabrachial nucleus (PB), the Kolliker-Fuse nucleus (KF), the spinal trigeminal nucleus caudalis (Sp5C), the nucleus of the solitary tract (NTS) and the ventrolateral medulla (VLM), which are activated by evoking the nasotrigeminal reflex and which exhibit immunoreactivity for the N-methyl-D-aspartate (NMDA) receptor subunit NR1. By stimulating the nasal mucosa with saline, we induced the expression of the immediate early gene c-fos and combined the immunocytochemical detection of the Fos protein with the detection of the NR1 subunit. Cell counts revealed that nasal stimulation, compared to anesthesia controls, resulted in highly significant increases (p < or = 0.001) of Fos-immunoreactive (-ir) neurons in the midlevel KF, the external lateral PB, and the Sp5C. In the central lateral PB, the rostral ventrolateral medulla including the Botzinger/pre-Botzinger complex, and in the ventrolateral and commissural NTS the increases were only moderately significant (p < or = 0.05). With respect to the numbers of NR1-/Fos-ir double labeled neurons, significant increases were only observed in a subset of these pontomedullary nuclei. Increases were highly significant in the Sp5C (p < or = 0.001) and the midlevel KF (p < or = 0.01) and moderately significant (p < or = 0.05) in the external lateral PB, Botzinger/pre-Botzinger complex, and ventrolateral NTS. The present study revealed that nasotrigeminally activated neurons in mandatory and potential relay sites of the nasotrigeminal reflex circuit express the NR1 subunit. This finding strongly suggests that NMDA-type glutamate receptors are involved in the mediation of the nasotrigeminally evoked cardiovascular and respiratory responses. PMID- 9853115 TI - Activity-dependent survival and enhanced turnover of calcium in cultured rat cerebellar granule neurons. AB - Neurons survive when their activity is maintained. An influential hypothesis on the cellular mechanism underlying this phenomenon is that there is an appropriate range of intracellular Ca2+ concentration ([Ca2+]i) for survival. The rat cerebellar granule neuron in culture serves as the most often used model system for the analysis of activity-dependent survival, since it does not survive unless an excitant (KCl or glutamate) is added to the culture medium. Against the above mentioned hypothesis, we found in our previous examination no difference between steady-state [Ca2+]i in granule neurons cultured under high KCl (i.e., survival) and low KCl (i.e., death) conditions. In this report, we present the quantitative background of unchanged [Ca2+]i between the two culture conditions. Influx of Ca2+ due predominantly to L-type voltage-dependent calcium channels was higher in high KCl cultures than in low KCl cultures. At the same time, efflux of Ca2+ due to the activity of Ca2+/Na+ antiport was also higher in high KCl cultures. Additionally, we found that the endocytotic activity was greater in high KCl cultures than in low KCl cultures, as monitored by the rate of uptake of horseradish peroxidase added to medium. Since the uptake was blocked by an internal Ca2+ chelator, the increased endocytotic activity in high KCl cultures might be a consequence of the enhanced Ca2+ turnover. PMID- 9853116 TI - Neuronal nicotinic acetylcholine receptors in rat trigeminal ganglia. AB - The application of nicotine to the various epithelia served by the trigeminal nerve produces irritation and/or pain by activating neuronal nicotinic acetylcholine receptors (NnAChRS) in sensory neurons. In this study the NnAChRs were identified in rat trigeminal ganglia (TG) using RT-PCR and immunocytochemistry. With RT-PCR the subunits of NnAChRs in rat TG were determined, and with immunocytochemistry the localization of three prominent subunits (alpha 7, alpha 4 and beta 2) were localized in intact TG neurons. The relative abundance of the alpha and beta subunits were: alpha 7 approximately alpha 3 > alpha 6 > alpha 4 approximately alpha 5 > alpha 9 > or = alpha 2, and beta 2 approximately beta 3 > beta 4. This is the first report of the alpha 9 subunit in TG. Immunohistochemical studies revealed that almost all TG neurons contained alpha 7-LI and alpha 4-LI, and that 85% had beta 2-LI. For these three subunits much of the label was internalized. Immunocytochemical studies using antibodies raised against chick alpha 8 subunits did not specifically label rat TG. These data reveal that rat TG neurons contain the entire spectrum of mammalian NnAChR subunits. PMID- 9853117 TI - The influence of removing extracellular Ca2+ in the desensitization responses to capsaicin, zingerone and olvanil in rat trigeminal ganglion neurons. AB - Desensitization is a process that describes the diminishing effect of a drug upon repeated applications. In regard to capsaicin, the pungent compound in hot pepper, it is well established that removal of extracellular calcium markedly diminishes desensitization. To explore whether this behavior extends to other analogues of capsaicin, we have determined the effect of removing extracellular calcium with capsaicin analogues, zingerone and olvanil, by whole-cell patch clamping cultured rat trigeminal ganglion neurons. Zingerone, like capsaicin, is pungent but has a shorter acyl chain, whereas olvanil is non-pungent and has a longer acyl chain. The currents evoked by 30-s applications of 30 mM zingerone or 1 microM olvanil repeated every 3 min differ in two important ways from the responses evoked by 1 microM capsaicin under these same conditions. In the presence of extracellular calcium, repeated applications of zingerone and olvanil produce nearly complete desensitization. Also in contrast to capsaicin, removing extracellular calcium for these two agonists does not diminish desensitization. These data analyses suggest the existence of calcium-independent pathways that can result in desensitization, and that pungency is not related to the phenomenon of desensitization. PMID- 9853118 TI - The origin and development of the vagal and spinal innervation of the external muscle of the mouse esophagus. AB - Retrograde and anterograde tracing and immunohistochemical techniques were used to examine the origin of the extrinsic innervation, and the development of the vagal innervation to the mouse esophagus. Cholinergic nerve terminals were localised using an antiserum to the vesicular acetylcholine transporter and cholinergic cell bodies were localised using an antiserum to choline acetyltransferase. Cholinergic nerve terminals, which also contained calcitonin gene-related peptide, were present at the motor end plates in the external (striated) muscle of the esophagus. Following injection of Fast Blue into subdiaphragmatic or cervical levels of the esophagus, the only retrogradely labelled cholinergic nerve cell bodies that also contained calcitonin gene related peptide were found in the nucleus ambiguus. Neurons in the dorsal motor nucleus of the vagus, the nodose ganglia and dorsal root ganglia gave rise to a number of different types of nerve terminals within the myenteric plexus. Retrogradely-labelled neurons in the dorsal motor nucleus of vagus contained cholinergic markers only, nitric oxide synthase only or cholinergic markers plus nitric oxide synthase, retrogradely-labelled neurons in the dorsal root ganglia contained calcitonin gene-related peptide only, and a small number of retrogradely-labelled neurons in the nodose ganglia contained tyrosine hydroxylase. The development of the vagal innervation to the esophagus was examined following application of DiI to the vagus nerve of fixed mouse embryos. Anterogradely-labelled nerve fibres, which arose from both nodose ganglia and the medulla, were already present in the esophagus of embryonic day 12 (E12) mice. Some of the DiI-labelled vagal nerve fibres were present in among the smooth muscle cells of the external muscle layer prior to their transdifferentiation to striated muscle. We conclude that the neurons in the nucleus ambiguus that project to the esophagus differ from other extrinsic neurons in their chemistry as well as their targets within the esophagus. The development of the extrinsic innervation precedes the transdifferentiation of the external muscle to striated muscle, raising the possibility that, during development, smooth muscle of the esophagus is innervated transiently by vagal neurons. PMID- 9853119 TI - Effect of low-intensity 50-Hz magnetic fields on kindling acquisition and fully kindled seizures in rats. AB - The possibility that chronic exposure to power-line frequency (50 Hz) magnetic fields (MFs) might affect the acquisition or characteristics of focal and generalized seizures in amygdala kindled rats was studied. Acute, short-lasting 50-Hz MF exposure of fully kindled rats at either 1 or 100 microT had no effect on afterdischarge threshold (ADT) or seizure parameters recorded at ADT. In the chronic experiments, rats with electrodes implanted in the basolateral amygdala were exposed to a 50-Hz, 100-microT (1 Gauss) MF or to a sham field condition before and after onset of daily electrical stimulations over the whole period of kindling development. The focal seizure threshold (ADT) was determined before and after kindling development in MF exposed and sham exposed rats. Pre-kindling ADT was significantly increased by MF exposure. Exposed rats needed about the same number of stimulations to kindle than sham exposed rats, but the cumulative afterdischarge duration to reach criterion (i.e., a stage 5 seizure) was significantly reduced in MF exposed animals. Post-kindling ADT was similar in the two groups, but MF exposed rats showed a significantly higher threshold for generalized seizures. The data indicate that chronic exposure of rats to a 50-Hz, 100-microT MF exerts weak inhibitory effects on some seizure parameters of the kindling model. PMID- 9853120 TI - Astrocytosis and amyloid deposition in scrapie-infected hamsters. AB - In scrapie infection, prion protein (PrPSc) is localized in areas where there is neurodegeneration and astrocytosis. It is thought that PrPSc is toxic to neurons and trophic for astrocytes. In our study, paraffin sections from scrapie infected (263K and 139H) and control hamsters were examined with histological and immunocytochemical staining. We found that PrPSc was present in the ependymal cells of both 263K- and 139H-infected hamsters. In 139H-infected hamsters, PrPSc was found in the cytoplasm of neurons in cerebral cortex and in hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. In contrast, neuronal cytoplasm and nuclei, were positive for PrPSc in most areas such as cortex, hippocampus, and thalamus in 263K-infected hamsters. Many aggregations of PrPSc could be seen in the cortex, hippocampus, substantia nigra and around the Pia mater, corpus callosum, fimbria, ventricles, and blood vessels in sections from 139H- and/or 263K-positive animals. Furthermore, PrPSc was also co-localized with glial fibrillary acidic protein (GFAP) in many reactive astrocytes (approximately 90%) in certain areas such as the hippocampus in 263K-infected hamsters, but not 139H-infected hamsters. The patterns of astrocytosis and PrPSc formation were different between 139H- and 263K-infected hamsters, which may be used for a diagnosis purpose. Our results suggest a hypothesis that multiple cell-types are capable of PrPSc production. Our results also confirm that reactive astrocytes can produce and/or accumulate PrPSc during some scrapie strain infections. The findings suggest a 'snowball effect', that is: astrocytosis might play an important role in amyloidosis, while amyloidosis may induce further astrocytosis at least in 263K-infected hamsters. PMID- 9853121 TI - Persistence and atrophy of septal/diagonal band neurons expressing the p75 neurotrophin receptor in pilocarpine-induced chronic epilepsy in the rat. AB - Systemic administration of pilocarpine, which results in status epilepticus followed by recurrent seizures in rats, is a widely used experimental model of chronic limbic epilepsy. Marked structural alterations have been documented in pilocarpine-induced epilepsy, and these include cell loss in the hippocampus and other brain areas, and sprouting of mossy and cholinergic fibers in the hippocampus. Evidence is accumulating that neurotrophins and neurotrophin receptors are involved in the cascade of these events. Two and 4 months after pilocarpine-induced epilepsy, neurons containing the 75-kDa low affinity neurotrophin receptor (p75NTR) were investigated with immunohistochemistry in the medial septal and diagonal band nuclei. No significant differences in the distribution and number of immunoreactive neurons were found in the epileptic rats compared to control saline-treated animals. However, in the epileptic animals, a significant decrease in the perikaryal size of p75NTR-immunoreactive neurons of the septal/diagonal band region was found by 60 days, and such atrophic changes were more marked in the diagonal band nuclei by 120 days. These findings indicate that the p75NTR-containing cell bodies, which include the neurons projecting to the hippocampal formation and are cholinergic in the normal brain, survive after months of spontaneous recurrent seizures, during which, therefore, a supply of p75NTR to target regions is maintained in the chronic epileptic brain. However, the present data point out that these p75NTR-containing neurons undergo a significant shrinkage in pilocarpine-induced chronic epilepsy, thus indicating that they are involved in the brain pathology of temporal lobe epilepsy. PMID- 9853122 TI - A microdialysis study investigating the mechanisms of hydroxyl radical formation in rat striatum exposed to glutamate. AB - Considerable evidence has linked hydroxyl radicals (.OH) to excitotoxicity. Glutamate infused through a microdialysis probe into rat striatum induced a massive .OH production, which was completely blocked by PBN and attenuated by dizocilpine, 2-amino-5-phosphonopentanoic acid (AP-5), NG-nitro-L-arginine methyl ester (L-NAME) and mepacrine. Thus, we suggest that the neurotoxic effects of glutamate in vivo may derive from an increased formation of .OH resulting from excessive activation of NMDA receptors and downstream enzymes such as NOS and PLA2. PMID- 9853123 TI - Adenosine and ADP prevent apoptosis in cultured rat cerebellar granule cells. AB - Cerebellar granule cells (CGCs) explanted in vitro undergo death via apoptosis when the concentration of potassium is shifted from 25 mM to 5 mM. We report that adenosine and ADP, which act as neurotransmitters and neuromodulators in the brain, exert in cultured cerebellar granule cells a specific and marked antiapoptotic action with half-maximal effect in the 10-100 microM range. The action of adenosine is partly inhibited by the A1AR antagonist 8-cyclopentyl-1,3 dipropylxanthine (DPCPX) and is mimicked by the A1AR agonist 2-chloro-N6 cyclopentyladenosine (CCPA), while ADP effect, that is completely blocked by the P2x, P2y receptors noncompetitive antagonist suramine, is restored in the presence of the selective P2x purinoceptors agonist beta, gamma-methylene-L-ATP. These findings demonstrate that adenosine and ADP markedly inhibit the program of cell death in cerebellar granule cells and suggest that such an action is mediated via interaction with, respectively, A1 and P2x receptors. PMID- 9853124 TI - Feeding effects of hypothalamic injection of melanocortin 4 receptor ligands. AB - It has been reported that intraventricular administration of the melanocortin 4 receptor (MC4-R) agonist MT II and antagonist SHU9119 alter food intake. We found that MT II and SHU9119 have extremely potent effects on feeding when injected in the paraventricular nucleus (PVN), a site where MC4-R gene expression is very high. Our finding provides direct evidence that MC4-R signaling is important in mediating food intake and that melanocortin neurons in the PVN exert a tonic inhibition of feeding behavior. Chronic disruption of this inhibitory signal is a possible explanation of the agouti-obesity syndrome. PMID- 9853125 TI - Brainstem carbachol injections in the urethane anesthetized rat produce hippocampal theta rhythm and cortical desynchronization: a comparison of pedunculopontine tegmental versus nucleus pontis oralis injections. AB - Previous research has demonstrated that brainstem injections of acetylcholine agonists (e.g., carbachol) produced electrophysiological indicators of rapid-eye movement (REM) sleep in the cat. Recent reports now indicate that this phenomenon may hold true for rats as well. Relatively few reports, however, have examined the effect of these injections on REM indicators in the anesthetized rat, a preparation useful for elucidating underlying neurobiological mechanisms controlling REM sleep processes. The present study compared the effect of injections of carbachol (5 micrograms in 250 nl) into the pedunculopontine tegmental nucleus (PPTg) or the nucleus pontis oralis (NPO) on two tonic indicators of REM sleep in the urethane-anesthetized rat. Namely, changes in the hippocampal EEG and in the cortical EEG. Carbachol injections into either site produced a change in both the hippocampal EEG and cortical EEG to a REM-like state at short latencies. The length of these changes (duration of effect), however, was site-dependent. Thus, PPTg carbachol injections induced significantly longer lasting effects in both the hippocampal and cortical EEG than did NPO injections. The results that brainstem carbachol injections in rats, as in cats, may provide a useful model for investigating tonic REM sleep processes. PMID- 9853126 TI - Morphological changes in the hippocampal CA3 region induced by non-invasive glucocorticoid administration: a paradox. AB - Repeated stress induces atrophy, or remodeling, of apical dendrites in hippocampal CA3 pyramidal neurons. In rats, the stress effect is blocked by adrenal steroid synthesis inhibitors, and mimicked by daily injection of corticosterone. We report that non-invasive administration of corticosterone in the drinking water (400 micrograms/ml) also produced atrophy of apical dendrites in CA3. Unexpectedly, the combination of daily stress and oral corticosterone negated the effects of either treatment alone, and no changes in the apical dendritic length or branching pattern of CA3 pyramidal neurons were observed compared to control unstressed rats. PMID- 9853127 TI - Mild kainate toxicity produces selective motoneuron death with marked activation of CA(2+)-permeable AMPA/kainate receptors. AB - Motoneuron death could be produced by higher sensitivity to excitoxicity during the development and pathological conditions. We report here that in ventral spinal cord cultures mild kainate exposure (12.5 microM, 20 min or 100 microM, 2.5 min) induced selective cobalt stain of motoneurons, indicating a specific Ca2+ entry through the Ca(2+)-permeable AMPA/kainate receptors. This result was associated with a selective motoneuron death as previously described. In these cultures, motoneuron immunoreactivity for the Ca2+ buffering protein, calretinin was negative. These findings suggest that the selective motoneuron death due to a mild excitotoxic insult could be linked to a marked Ca2+ influx associated with the lack of some Ca2+ buffering proteins. PMID- 9853128 TI - Light-increased cGMP and K+ conductance in the hyperpolarizing receptor potential of Onchidium extra-ocular photoreceptors. AB - The phototransduction mechanism of the extra-ocular photoreceptor cells Ip-2 and Ip-1 in the mollusc Onchidium ganglion was examined. Previous work showed that the depolarizing receptor potential of another extra-ocular photoreceptor cell, A P-1 is produced by a decrease of the light-sensitive K+ conductance activated by a second messenger, cGMP and is inactivated by the hydrolysis of cGMP. Here, a hyperpolarizing receptor potential of Ip-2 or Ip-1 was associated with an increase in membrane conductance. When Ip-2 or Ip-1 was voltage-clamped near the resting membrane potential, light induced an outward photocurrent corresponding to the above hyperpolarization. The spectral sensitivity had a peak at 510 nm. The shift of reversal potentials of the photocurrent depended on the Nernst equation of K(+)-selective conductance. The photocurrent was blocked by 4-AP and L-DIL, which are effective blockers of the A-P-1 light-sensitive K+ conductance. These results suggested that the hyperpolarization is mediated by increasing a similar light-sensitive K+ conductance to that of A-P-1. The injection of cGMP or Ca2+ into a cell produced a K+ current that mimicked the photocurrent. 4-AP and L DIL both abolished the cGMP-activated K+ current, while TEA suppressed only the Ca(2+)-activated K+ current. These results indicated that cGMP is also a second messenger that regulates the light-sensitive K+ conductance. The photocurrent was blocked by LY-83583, a guanylate cyclase (GC) inhibitor, but was unaltered by zaprinast, a phosphodiesterase (PDE) inhibitor. Together, the present results suggest that increasing the internal cGMP in Ip-2 or Ip-1 cells light-activates GC rather than inhibits PDE, thereby leading to an increase of the light sensitive K+ conductance and the hyperpolarization. PMID- 9853129 TI - The Janeway Lecture. Gynecologic oncology in the last quarter century. PMID- 9853130 TI - Lymphatic mapping and sentinel node biopsy: a new era in the management of solid neoplasms? PMID- 9853131 TI - Prostate cancer: patient's dilemma? PMID- 9853132 TI - How to do more with less. PMID- 9853133 TI - Universal application of intraoperative lymphatic mapping and sentinel lymphadenectomy in solid neoplasms. AB - PURPOSE: Regional lymph node involvement is the most important prognostic indicator in patients with solid tumors. Conventional lymph node dissection has not been shown to affect survival and is often associated with considerable morbidity. Intraoperative lymphatic mapping and sentinel lymph node dissection were therefore designed as a minimally invasive alternative to routine elective lymph node dissection in patients with primary cutaneous melanoma. This study examined whether introperative lymphatic mapping and sentinel lymph node dissection were accurate in staging patients with other solid malignancies. PATIENTS AND METHODS: Between 1985 and 1998, 107 patients with breast cancer, 17 with thyroid tumors, 14 with gastrointestinal/gynecologic cancers, six with Merkel cell cancers, and five with squamous cell carcinomas of the head and neck have undergone mapping and sentinel lymph node dissection at the John Wayne Cancer Institute. RESULTS: The sentinel node was identified in 96% of patients (98% melanoma). In 36% of patients the sentinel node was the only tumor-positive node (71% melanoma). Eighteen percent of sentinel nodes were negative by hematoxylin and eosin staining but were positive by immunohistochemical staining (15% melanoma). CONCLUSION: These data suggest that many solid neoplasms have a primary lymphatic channel and lymph node to which it drains. Although sentinel lymph node dissection has been popularized in melanoma therapy, we have found it feasible for treatment of other solid malignancies. This technique may ultimately replace conventional dissection with more accurate staging. PMID- 9853134 TI - Simultaneous radiotherapy for prostate cancer: 125I prostate implant followed by external-beam radiation. AB - PURPOSE: Using a rigorous prostate-specific antigen definition of disease freedom, the 10-year disease-free survival rates after simultaneous radiation of prostate cancer are presented. PATIENTS AND MATERIALS: From January 1984 through December 1996, 1020 men with clinical stage T1T2N0 prostate cancer were treated by simultaneous radiation: radioactive 125I prostate implantation followed by external-beam radiation. The median pretreatment prostate-specific antigen was 7.5 ng/mL (range, 0.2-188 ng/mL). Implantation was performed by both the retropubic and the transperineal technique, always followed by external-beam radiation. None received hormone treatment. Disease freedom is defined as achieving and maintaining a posttreatment prostate-specific antigen of < or = 0.5 ng/mL. The median follow-up is 3 years (range, 1-14 years). RESULTS: The overall 5- and 10-year disease-free survival rates are 79% and 72%, respectively, after which a plateau is reached. At 5 years posttreatment, significantly better disease-free survival results are documented with simultaneous radiation by the ultrasound technique (92%) compared with the retropubic implant technique (73%). On multivariate analysis, pretreatment prostate-specific antigen is the most significant factor associated with disease-free survival, followed by implant technique. DISCUSSION: The 10-year disease-free survival rate after simultaneous radiation is comparable to the 10-year results after radical prostatectomy. Disease freedom is defined by the same prostate-specific antigen criteria used for surgery. A plateau in the disease-free curve suggests cure. Of equal importance, the information described in this report should form only a baseline relative to future results as men treated with simultaneous radiation using the transperineal implant technique reach longer follow-up. PMID- 9853135 TI - Selective radiosensitization of 9L glioma in the brain transduced with double suicide fusion gene. AB - PURPOSE: Suicide gene therapy has proved to be successful in enhancing the therapeutic index by sensitizing genetically modified tumor cells to prodrugs. Two of the most widely studied suicide genes, herpes simplex virus type 1 thymidine kinase and Escherichia coli cytosine deaminase, have proved effective at selectively eliminating malignant tumor cells. We previously demonstrated that transduced 9L glioma cells expressing E. coli cytosine deaminase and herpes simplex virus type 1 thymidine kinase concomitantly as a fusion protein exhibited greater levels of targeted cytotoxicity and radiosensitization than could be achieved by single suicide gene therapy. The present in vivo studies were carried out to determine whether double suicide gene therapy would enhance the tumor control rate of orthotopically implanted malignant glioma growing in the brain when coupled with radiotherapy. MATERIALS AND METHODS: Rat 9L gliosarcoma cells were transfected with retroviral vectors containing an E. coli cytosine deaminase and herpes simplex virus type 1 thymidine kinase fusion gene and maintained in Dulbecco's modified Eagle's medium. The antitumor response of 9L E. coli cytosine deaminase and herpes simplex virus type 1 thymidine kinase tumors growing in the brain of Fischer rats was evaluated with small tumors (6-day-old tumors) versus large tumors (14-day-old tumors) against single versus double prodrug treatments. In the large brain tumors, the therapeutic efficacy of the combined single and double prodrugs coupled with radiotherapy was evaluated. RESULTS: Double suicide gene therapy using two prodrugs, 5-fluorocytosine (500 mg/kg) and ganciclovir (30 mg/kg), was effective in achieving long-term tumor control (50% survival) against early-stage brain tumors (6 days after implantation) but was only marginally effective against advanced stage tumors (14 days old). However, when these prodrugs were combined with radiotherapy and double suicide gene therapy against advanced-stage tumors, more than 70% of the animals were cured, whereas radiotherapy alone (20 Gy) failed to achieve any cure at all. Combined radiotherapy and single prodrug therapy showed a moderate increase in the animal survival rate (17% and 40% for 5-fluorocytosine and ganciclovir, respectively) but was inferior to the combination therapy of radiation and double prodrugs. CONCLUSION: The present in vivo results indicate that double suicide gene therapy combined with radiotherapy may represent a new, effective approach to achieve a high tumor cure rate without producing any excessive normal tissue damage. PMID- 9853136 TI - Identification of a high-risk clinically localized prostate cancer subgroup receiving maximum benefit from whole-pelvic irradiation. AB - PURPOSE: We recently identified a progression-free survival advantage for clinically localized high-risk prostate cancer patients receiving whole-pelvic irradiation. We now seek to identify a subgroup most likely to benefit from whole pelvic irradiation. METHODS: Between October 1987 and December 1995, 506 clinically localized prostate cancer patients were treated with definitive radiotherapy consisting of whole-pelvic irradiation followed by a prostate-only boost, or prostate-only treatment (median follow-up, 35 months vs 30 months). Prostate-specific antigen (PSA) failure was defined as (1) a PSA value > or = 1 ng/mL or (2) a PSA value that rose > or = 0.5 ng/mL in < or = 1 year posttreatment on two consecutive measurements, with the first rise defined as the time of failure. The calculated risk of lymph node positivity (%rLN+) was defined as 2/3 (initial PSA) + 10(Gleason score - 6), with intermediate risk defined as 15% < or = %rLN+ < 35% and highest risk defined as %rLN+ > or = 35%. Univariate and multivariate analyses were performed. RESULTS: Intermediate-risk patients receiving whole-pelvic irradiation had significantly improved freedom from PSA failure compared with those receiving prostatic irradiation only (median progression-free survival 39.5 months vs 22.5 months; P < 0.0001); highest-risk patients did not (median progression-free survival 27.2 months vs 20.8 months, P = NS). Multivariate analysis revealed type of radiation treatment to be the most significant independent predictor of outcome (P < 0.0001). DISCUSSIONS: Whole pelvic radiotherapy most significantly improves the PSA failure-free survival in patients with an intermediate calculated risk of lymph node positivity, suggesting that highest-risk patients may present with distant micrometastases. PMID- 9853137 TI - A phase II/III clinical study of tin ethyl etiopurpurin (Purlytin)-induced photodynamic therapy for the treatment of recurrent cutaneous metastatic breast cancer. AB - BACKGROUND: Chest wall recurrence of breast cancer after mastectomy, radiation therapy, and chemotherapy poses a therapeutic dilemma. Further intervention with any or all of these modalities is often futile and morbid. Left untreated, severe pain, infection, and suffering occur. OBJECTIVE: To ascertain whether photodynamic therapy may present a palliative option for these individuals. METHODS: A total of 86 lesions (2.4-cm mean diameter) were treated on eight patients who had biopsy-proven chest wall recurrence despite surgery, chemotherapy, and radiation therapy. Each patient underwent a single photodynamic therapy session in which 1.2 mg/kg of the drug tin ethyl etiopurpurin (Purlytin) was injected and followed 24 hours later by laser light treatment at 660 +/- 3 nm (at 150 mW/cm2 for a total light dose of 200 J/cm2). RESULTS: With a minimum 6 month follow-up, the objective response rates after photodynamic therapy were complete response, 92%; partial response, 8%; and no response, 0%. Lesions less than 0.5 cm had a 100% complete response. Morbidity was minimal with no systemic toxicity. One patient had a wound infection that responded to oral antibiotics. No photosensitivity reactions were reported in this set of patients. Posttreatment pain was reported and could be treated with medication and application of cold compresses. CONCLUSIONS: Photodynamic therapy offers an excellent local control rate of chest wall recurrence with minimal morbidity after multimodality treatment failure. The treatment is given in a single session and on an outpatient basis. In patients who may register a partial response or have recurrence or the incidence of further chest wall nodules after photodynamic therapy, the treatment is repeatable. PMID- 9853138 TI - A preliminary report: frequency of A-T heterozygotes among prostate cancer patients with severe late responses to radiation therapy. AB - PURPOSE: To investigate whether a significant proportion of prostate cancer patients who have late sequelae after high-dose external-beam conformal radiation therapy are radio-sensitive because they are carriers of ataxia-telangiectasia, that is, are heterozygous for mutations in the ATM gene. PATIENTS AND METHODS: A group of prostate cancer patients were selected who experienced severe late sequelae, specifically proctitis or cystitis, after high-dose external-beam conformal radiation therapy, together with a control group of patients treated in the same way but who did not have severe late effects. Blood samples were taken from these patients, genomic DNA extracted, and mutations sought in the ATM gene. RESULTS: Of 17 late-effect patients in whom most or all of the ATM gene has been examined, significant mutations (17.6%) were identified in three. No significant mutations were found in the control group. The incidence of ataxia- telangiectasia heterozygotes in the United States population is 1% to 2%. DISCUSSION: These preliminary data suggest that a disproportionate number, but by no means all, of prostate cancer radiotherapy patients who experience severe late effects are ataxia-telangiectasia heterozygotes. If this conclusion is confirmed, these individuals could be identified prospectively and, with dose de-escalation, spared a great deal of discomfort and suffering. As a corollary, if most of the small late-effects population were prospectively identifiable, the dose to the remaining population could potentially be escalated. Present methods of identifying mutations in a large gene, such as ATM, are cumbersome and expensive, but the technology is evolving rapidly, so that rapid screening of the ATM gene is imminent. PMID- 9853139 TI - Uterine artery embolization for fibroid disease. PMID- 9853140 TI - Complications of lower-limb percutaneous transluminal angioplasty: a prospective analysis of 410 procedures on 295 consecutive patients. AB - PURPOSE: To evaluate complications and their predictors in percutaneous transluminal angioplasty (PTA) of lower-limb arteries. METHODS: Complications in 410 angioplasty procedures in 295 consecutive patients (192 claudicants and 103 suffering from chronic critical ischemia) were prospectively analyzed. RESULTS: The total complication rate was 10.5% (43/410). There were 21 major complications (5%), eight of which required surgical treatment, including four hematomas, two arteriovenous fistulae, and two pseudoaneurysms at the puncture site, two retroperitoneal hematomas, and 11 thrombotic/thromboembolic complications. There were significantly more complications with treatment of occlusions compared with stenoses (18% vs 7%, p = 0.002). Women had significantly more bleeding complications than men (15% vs 6%, p = 0.032). The 30-day mortality rate in patients with critical ischemia was 10%. CONCLUSION: In lower-limb PTA a few target lesion- and patient-related determinants of complications could be identified. In patients with critical ischemia, the 30-day mortality was rather high and mainly due to associated coronary and cerebrovascular diseases. PMID- 9853141 TI - Comparison of angiography and intravascular ultrasound before and after balloon angioplasty of the femoropopliteal artery. AB - PURPOSE: To compare angiographic and intravascular ultrasound (IVUS) data before and after balloon angioplasty (PTA) of the femoropopliteal artery. METHODS: Qualitative and quantitative analyses were performed on corresponding angiographic and IVUS levels obtained from 135 patients. RESULTS: IVUS detected more lesions, calcified lesions, and vascular damage than angiography. Sensitivity of angiography was good for the presence of a lesion (84%), moderate for eccentric lesions (53%) and for vascular damage (52%), and poor for calcified lesions (30%). The increase in angiographic diameter stenosis was associated with a decrease in lumen area and increase in percentage area stenosis on IVUS. CONCLUSIONS: Angiography is less sensitive than IVUS for detecting lesion eccentricity, calcified lesions, and vascular damage. Presence of a lesion and amount of plaque were underestimated angiographically. Only before PTA was good agreement found between angiographic diameter stenosis and lumen size on IVUS. PMID- 9853142 TI - Low-molecular-weight heparin (reviparin) reduces the incidence of femoropopliteal in-stent stenosis: preliminary results of an ongoing study. AB - PURPOSE: To examine the efficacy of the low-molecular-weight heparin, reviparin, for prevention of femoropopliteal stent restenosis. METHODS: Forty-two patients who had implantation of flexible tantalum stents for the treatment of stenosis (n = 24) or occlusion (n = 18) of the femoral (n = 27) or popliteal (n = 15) arteries were included in this study protocol. An intraarterial bolus of 5000 IU heparin was given before percutaneous transluminal angioplasty (PTA), and in the case of stent implantation due to unsuccessful PTA, an additional dose of reviparin (3500 anti-factor Xa IU) was given. Postprocedurally, 10,500 anti factor Xa IU of reviparin were administered intravenously over 24 hr, followed by 3500 anti-factor Xa IU subcutaneously twice a day for 23 days. Oral aspirin (100 mg/day) was prescribed for the long term. Follow-up criteria (maximum follow-up 37 months) were clinical symptoms, Doppler ankle arm indices, color and duplex sonography, and angiography for suspicion of restenosis. RESULTS: Early stent thromboses were not observed. Overall primary patency rate (PPR) was 88% +/- 6.0% (1 year) and 74% +/- 10.1% (2 years). Major hemorrhagic complications have not occurred. CONCLUSION: Reviparin administered in a high dose over a period of 24 days is a safe medication regimen and provides excellent patency rates after stent implantation. PMID- 9853143 TI - Renal artery stenosis: changes in intrarenal Doppler waveform following percutaneous transluminal angioplasty. AB - PURPOSE: To examine the extent of the changes in the intrarenal spectral waveform patterns after percutaneous transluminal angioplasty (PTA) and whether there is a correlation with the angiographic and clinical results. METHODS: In 44 patients with 68 PTAs we analyzed the intrarenal spectral waveform regarding the existence of a tardus-parvus pattern before and after PTA. RESULTS: In 51 of the 60 cases with a tardus-parvus pattern prior to PTA, a complete normalization of the spectral wave-form was noted. There was no correlation between the Doppler result after PTA and the angiographic and clinical result. In contrast there was a significant correlation between the Doppler result before PTA and the clinical outcome: patients with a normal intrarenal spectral waveform before PTA showed no improvement in their arterial hypertension. CONCLUSION: Our results indicate that a patient with a normal spectral waveform analysis does not response to PTA. However, there is still an unpredictable clinical response even if a patient has an abnormal intrarenal spectral waveform prior to PTA and a complete normalization after PTA. PMID- 9853144 TI - Primary stenting for complex atherosclerotic plaques in aortic and iliac stenoses. AB - PURPOSE: To evaluate the efficacy of primary stenting for complex atherosclerotic plaques in aortic and iliac stenoses that are not amenable to balloon angioplasty alone. METHODS: Nineteen patients with complex atherosclerotic plaques were treated with a Palmaz stent (n = 19), Wallstent (n = 1), Strecker stent (n = 1), or Memotherm stent (n = 1). A total of 22 stenoses presenting with complex plaque morphology including ulcerated plaques, ulcerated plaques with focal aneurysms, plaques with heavy calcification, severely eccentric plaques, plaques with overhanging edge, and plaques with spontaneous dissection were stented. The lesions were in the aorta (n = 1), common iliac artery (n = 19), or external iliac artery (n = 2). RESULTS: Immediate angiography after stent placement revealed restoration of patency of the stented segment. Focal aneurysms and ulcerated areas were occluded in the follow-up angiographies obtained 4-12 weeks after the procedure. In one case with poor distal runoff and multiple complex lesions of the iliac artery, subacute occlusion occurred. Clinical and angiographic follow-up (3-46 months) revealed patency of all other stented segments. CONCLUSION: Primary stenting is an effective and reliable approach for complex plaques in stenoses. Patency of the arterial segment with a smooth lumen can be created without the risk of acute complications such as distal embolization, dissection, or occlusion. PMID- 9853145 TI - Effect of preoperative radiological treatment of hepatocellular carcinoma before liver transplantation: a retrospective study. AB - PURPOSE: To assess the results of radiological treatment of patients with hepatocellular carcinoma (HCC) performed before orthotopic liver transplantation (OLT). METHODS: Sixty-two transplanted patients with a total of 89 HCC nodules were studied; 50 lesions in 38 patients had been treated prior to OLT with transcatheter arterial chemoembolization (TACE; n = 29), percutaneous ethanol injection (PEI; n = 10), or combined therapy (TACE + PEI; n = 11). The induced necrosis was pathologically evaluated. The recurrence rate after OLT in the treated group of patients (n = 38) was compared with that in the non-treated group (n = 24). RESULTS: After TACE, necrosis was complete in 7 of 29 lesions (24.1%), partial in 11 of 29 (37.9%), and absent in 11 of 29 (37.9%). After PEI, necrosis was complete in 8 of 10 lesions (80%), and partial in 2 of 10 (20%). Using combined therapy, necrosis was complete in 11 of 11 lesions (100%). Four of 24 untreated and 4 of 38 treated patients did not survive OLT from causes not related to the HCC; 3 of 20 non-treated patients (15%) and 4 of 34 treated patients (11.8%) had post-OLT recurrence (these last four patients had undergone only TACE and did not have tumor necrosis at pathological examination). CONCLUSION: TACE of HCC prior to OLT had no influence on the recurrence rate. PEI and combined therapy (TACE + PEI) may be recommended in patients awaiting OLT. PMID- 9853146 TI - Percutaneous mitoxantrone injection for primary and secondary liver tumors: preliminary results. AB - PURPOSE: To determine the effects of percutaneous intratumoral chemotherapy with mitoxantrone (PIM) in the palliative treatment of malignant liver lesions. METHODS: We treated 15 progressive lesions in nine patients in whom either previous therapy failed or serious complications developed as a result. Seven lesions were metastatic and eight were due to foci of hepatocellular carcinoma. Under computed tomography (CT) guidance, we percutaneously injected 10-20 mg of mitoxantrone mixed with 0.5 ml of contrast medium into the tumor, performing one to three treatments at intervals of 1 month. RESULTS: There were no complications. The morphologic responses of the tumors after treatment were: minor response in one case, no change in 11 cases, progressive disease in three cases. Mitoxantrone induced tumor necrosis with no viable cancer tissue in eight of 11 biopsies. Recurrence was observed in nine of the treated lesions 2-9 months after treatment. New lesions were observed in five of nine patients 1-9 months after treatment. CONCLUSION: In patients with malignant liver lesions with no other therapeutic possibilities, minimally invasive intratumoral mitoxantrone injection was carried out safely with good tumor delivery of chemotherapy, and tumor necrosis was demonstrated at biopsy. We feel this approach warrants further investigation. PMID- 9853147 TI - MR-guided percutaneous angioplasty: assessment of tracking safety, catheter handling and functionality. AB - PURPOSE: Magnetic resonance (MR)-guided percutaneous vascular interventions have evolved to a practical possibility with the advent of open-configuration MR systems and real-time tracking techniques. The purpose of this study was to assess an MR-tracking percutaneous transluminal angioplasty (PTA) catheter with regard to its safety profile and functionality. METHODS: Real-time, biplanar tracking of the PTA catheter was made possible by incorporating a small radiofrequency (RF) coil in the catheter tip and connecting it to a coaxial cable embedded in the catheter wall. To evaluate potentially hazardous thermal effects due to the incorporation of the coil, temperature measurements were performed within and around the coil under various scanning and tracking conditions at 1.5 Tesla (T). Catheter force transmission and balloon-burst pressure of the MR tracking PTA catheter were compared with those of a standard PTA catheter. The dilatative capability of the angioplasty balloon was assessed in vitro as well as in vivo, in an isolated femoral artery segment in a swine. RESULTS: The degree of heating at the RF coil was directly proportional to the power of the RF pulses. Heating was negligible with MR tracking, conventional spin-echo and low-flip gradient-echo sequences. Sequences with higher duty cycles, such as fast spin echo, produced harmful heating effects. Force transmission of the MR-tracking PTA catheter was slightly inferior to that of the standard PTA catheter, while balloon-burst pressures were similar to those of conventional catheters. The MR tracking PTA catheter functioned well both in vitro and in vivo. CONCLUSION: The in vivo use of an MR-tracking PTA catheter is safe under most scanning conditions. PMID- 9853148 TI - Single and tandem stents in sheep iliac arteries: is there a difference in patency? AB - PURPOSE: To compare patency and neointima formation of single and tandem arterial stents. METHODS: In each of six sheep, two Memotherm nitinol stents (tandem stents) were inserted into the external iliac artery on one side and a single stent into the artery on the opposite side. The size of the iliac lumen was assessed in the proximal, middle, and distal segments of the stents by intravascular ultrasound (IVUS) before, immediately after, and 1 month after implantation when the sheep were killed. Neointimal thickness was determined in the proximal, middle, and distal segments of each stent by light microscopy. RESULTS: All stents remained patent. There was no significant difference in lumen and neointimal thickness between single and tandem stents. Cranial tandem stents showed a significantly wider lumen and smaller neointimal thickness than caudal tandem stents. In the proximal and distal segments, the lumen of the stents was significantly smaller and the neointimal thickness greater than in the middle segment; differences in neointimal thickness were significant only between the proximal and the middle segment. CONCLUSION: In an experimental setting, tandem stents did not interfere with one another with regard to patency and neointima formation when compared with a single contralateral stent. Neointimal thickening after stent insertion seems to be inversely related to the original arterial diameter. PMID- 9853149 TI - Intracerebral hemorrhage after transcatheter thrombolysis of non-occluding superior mesenteric artery thrombosis. AB - We performed transcatheter thrombolysis on a 64-year-old man with non-occluding superior mesenteric artery (SMA) thrombosis because his severe symptoms could not be controlled with medication. An enhanced computed tomography (CT) scan revealed intramural thrombosis in the SMA. We were concerned that the narrowing of the SMA lumen might progress to complete occlusion, resulting in a high likelihood of mortality. After dissolution of the SMA thrombosis, the original symptoms almost completely disappeared. However, intracranial hemorrhage occurred 8 hr after thrombolysis, requiring surgical intervention. Transcatheter thrombolysis is thought to be a useful treatment for SMA thrombosis, especially in elderly patients with a high operative risk; however, the possibility of intracerebral hemorrhage must be taken into consideration. PMID- 9853150 TI - Congenital intrahepatic portosystemic shunt. AB - The authors report a congenital intrahepatic portosystemic shunt detected by angiography in a young patient with acute onset of hyperammoniemia and hepatic encephalopathy. PMID- 9853151 TI - Entrapment of J-tip guidewires by Venatech and stainless-steel Greenfield vena cava filters during central venous catheter placement: percutaneous management in four patients. AB - We present four patients in whom bedside placement of a central venous catheter was complicated by entrapment of a J-tip guidewire by a previously placed vena cava (VC) filter. Two Venatech filters were fragmented and displaced into the superior VC or brachiocephalic vein during attempted withdrawal of the entrapped wire. Two stainless-steel Greenfield filters remained in place and intact. Fluoroscopically guided extraction of both wires entrapped by Greenfield filters was successfully performed in the angiography suite. PMID- 9853152 TI - Delayed esophageal hemorrhage caused by a metal stent: treatment with embolization. AB - We report a case of life-threatening esophageal hemorrhage after metal stent implantation successfully treated by arterial embolization. An 85-year-old woman was admitted in shock secondary to massive hematemesis and melena. Recent medical history revealed esophageal cancer treated 8 weeks previously by endoesophageal radiotherapy (40 Gy) and endoscopic placement of a covered Wall-stent prosthesis. Selective arteriography of the fifth posterior right intercostal artery showed massive contrast extravasation in the esophagus. Embolization was performed with 150-250-micron polyvinyl alcohol particles. Follow-up at 5 months was uneventful. Arteriography and embolization are advised when severe hemorrhage occurs after esophageal implantation of metal stents. PMID- 9853153 TI - A balloon protection sheath to prevent peripheral embolization during aortoiliac endovascular procedures. AB - Peripheral embolization is a known complication occurring during aortoiliac recanalization. In particular, acute, fresh thrombotic occlusions, with a history shorter than 6 months, have a high risk for peripheral embolization. We describe a balloon occlusion sheath used to manage embolic material while preventing peripheral embolization. PMID- 9853154 TI - Percutaneous retrieval of a right atrioventricular embolus. AB - Percutaneous retrieval of a 12-cm-long serpiginous clot lodged in the right atrium and ventricle is reported. Following bilateral common femoral vein puncture, a Bird's Nest cava filter was first positioned ready to deploy immediately below the renal veins via the right femoral vein. From the left femoral vein, a Cook intravascular retrieval basket was advanced to the right atrium. Under transthoracic echocardiographic visualization, the basket was used to engage, trap, and gently withdraw the clot in a single long strand below the prepositioned inferior vena cava filter. The filter was immediately deployed, leaving the clot trapped inferior to the renal veins, in the cava and left iliac vein. The patient remained well and asymptomatic at discharge. PMID- 9853155 TI - Recanalization of obstructed Tenckhoff peritoneal dialysis catheter: wire/stylet manipulation combined with endoluminal electrocauterization. AB - We report the results of fluoroscopically guided wire/stylet manipulation combined with endoluminal electrocauterization in seven patients with obstructed Tenckhoff peritoneal dialysis catheters. In preparation for clinical application, electrocauterization was performed using a stone basket to recanalize surgically removed Tenckhoff catheters obstructed with omental fat ingrowing through the side holes. All ingrowing omental fat was removed easily by electrocauterization with the rotating movement of a stone basket. The technique was then applied in vivo in seven cases with ingrowing omental fat and malpositioned catheter; six (86%) were successfully recanalized. Among those six cases with initial success, four maintained good catheter function with durable patency (mean 261.3 days). No significant complication was noted. PMID- 9853156 TI - Pseudocoarctation of the aorta: complementary findings on plain film radiography, CT, DSA, and MRA. PMID- 9853157 TI - Re: Stent placement at the venous anastomosis of a dialysis graft: be cost effective and demanding with regard to the result. PMID- 9853158 TI - Plea for a "surgical conscience" in the interventional radiology suite. PMID- 9853159 TI - Surgical face masks: are they mandatory for interventional endovascular procedures? PMID- 9853160 TI - Nasolacrimal polyurethane stent: complications with CT correlation. AB - PURPOSE: To evaluate initial results in patients with epiphora secondary to obstruction of the nasolacrimal duct treated by placement of a polyurethane stent, and to discuss the technical problems and complications arising during the procedure, with visualization of the anatomy of the drainage apparatus using computed tomography (CT). METHODS: We inserted 20 polyurethane Song stents under fluoroscopic guidance after dacryocystography in 19 patients with grade 3-4 epiphora caused by idiopathic obstruction of the nasolacrimal duct. CT scans were obtained following stent placement in all patients. RESULTS: We focus on the technical problems and complications that arose during these procedures. During negotiation of the guidewire past the obstruction at the level of the junction of the duct with the lacrimal sac, the guidewire created a false passage in a posterior suborbital direction in two cases and towards the posterior midline in another. In all cases the guidewire was withdrawn and reinserted through the proper anatomic route without further difficulty or complications. In two cases the stent was improperly positioned wholly or partially outside the nasolacrimal system (one medially, one posteriorly). In one case the stent was removed and reinserted; in the other it remains in place and functional. CT was performed in all these cases to ensure proper anatomic alignment and determine what had gone wrong. The epiphora was completely resolved in 13 cases and partially relieved in four; there were three cases of stent obstruction. Epistaxis of short duration (1 hr) occurred in seven patients and headache in one. CONCLUSIONS: Treatment of epiphora with polyurethane stents is a technique that is well tolerated by patients and achieves a high success rate, yet problems in placement may be encountered. Though no major consequences for patients are involved, cognizance of such difficulties is important to avoid incorrect positioning of stents. PMID- 9853161 TI - Penetrating atherosclerotic ulcer of the descending thoracic aorta: treatment by endovascular stent-graft. AB - PURPOSE: To present four cases of penetrating ulcer of the descending thoracic aorta treated by transfemoral insertion of an endoluminal stent-graft. METHODS: Four patients with penetrating aortic ulcers were reviewed. Three cases were complicated by rupture, false aneurysm, or retrograde dissection. All patients were treated by endovascular stent-graft and were followed by helical computed tomography (CT). RESULTS: Endovascular stent-graft deployment was successful in all patients. However, in one case we observed a perigraft leak that spontaneously disappeared within the first month, and two interventions were needed for another patient. Following treatment, one episode of transient spinal ischemia was observed. The 30-day survival rate was 100%, but one patient died from pneumonia with cardiac failure 34 days after the procedure. In one patient, helical CT performed at 3 months showed a false aneurysm independent of the first ulcer. This patient refused any further treatment and suddenly died at home (unknown cause) after a 6-month follow-up period. CONCLUSION: Transluminal placement of endovascular stent-grafts for treatment of penetrating ulcers of the descending thoracic aorta appears to be a possible alternative to classical surgery. After treatment, follow-up by CT is essential to detect possible complications of the disease. PMID- 9853162 TI - Does coronary stenting following balloon angioplasty improve myocardial fractional flow reserve? AB - PURPOSE: Suboptimal distal coronary flow reserve after successful balloon angioplasty has been attributed to angiographically unrecognized inadequate lumen expansion, and adjunct coronary stenting has been shown to improve coronary flow reserve. The aim of this study was to investigate whether myocardial fractional flow reserve (FFRmyo) would increase further after coronary stenting compared with balloon angioplasty alone in the same patient group. METHODS: FFRmyo and quantitative coronary angiography were obtained before and after pre-stent balloon dilation, and again after stent placement in 11 patients (7 left anterior descending artery, 3 right coronary artery and 1 left circumflex artery). FFRmyo was calculated as the ratio of Pd/Pa during intracoronary adenosine 5' triphosphate (50 micrograms and 20 micrograms in the left and right coronary arteries, respectively)-induced maximum hyperemia, where Pd represents mean distal coronary pressure measured by a 2.1 Fr infusion catheter and Pa represents mean aortic pressure measured by the guiding catheter. RESULTS: Percent diameter stenosis significantly decreased after balloon angioplasty (74% +/- 15% vs 37% +/ 17%, p < 0.001), and decreased further after stent placement (18% +/- 10%, p < 0.001 vs baseline and balloon angioplasty). FFRmyo after coronary stenting (0.85 +/- 0.09) was significantly higher than that at baseline (0.51 +/- 0.16, p < 0.001) and after balloon angioplasty (0.77 +/- 0.11, p < 0.05). There was a significant correlation between angiographic variables and FFRmyo. The increase in lumen dimensions after coronary stenting was followed by a further significant improvement of FFRmyo. CONCLUSION: These results suggest that coronary stenting may provide a more favorable functional status and lumen geometry of residual coronary stenosis compared with balloon angioplasty alone. PMID- 9853163 TI - Indications, management, and complications of temporary inferior vena cava filters. AB - PURPOSE: We describe the results of a preliminary prospective study using different recently developed temporary and retrievable inferior vena cava (IVC) filters. METHODS: Fifty temporary IVC filters (Gunther, Gunther Tulip, Antheor) were inserted in 47 patients when the required period of protection against pulmonary embolism (PE) was estimated to be less than 2 weeks. The indications were documented deep vein thrombosis (DVT) and temporary contraindications for anticoagulation, a high risk for PE, and PE despite DVT prophylaxis. RESULTS: Filters were removed 1-12 days after placement and nine (18%) had captured thrombi. Complications were one PE during and after removal of a filter, two minor filter migrations, and one IVC thrombosis. CONCLUSION: Temporary filters are effective in trapping clots and protecting against PE, and the complication rate does not exceed that of permanent filters. They are an alternative when protection from PE is required temporarily, and should be considered in patients with a normal life expectancy. PMID- 9853164 TI - Percutaneous transluminal angioplasty and enclosed thrombolysis versus percutaneous transluminal angioplasty in the treatment of femoropopliteal occlusions: results of a prospective randomized trial. AB - PURPOSE: To determine whether percutaneous transluminal angioplasty (PTA) and enclosed thrombolysis (ET) is superior to PTA alone in the treatment of femoropopliteal occlusions. METHODS: Twenty-five patients with 5-15-cm-long occlusions in the femoropopliteal segments, with otherwise normal run-in arteries and at least one normal tibioperoneal artery to the foot, were randomized to ET/PTA or PTA alone. Ankle brachial systolic index (ABI) was measured before the procedure and at 24 hr and 12 months after the procedure, when a duplex scan was also carried out. End points in the study were patency at, or repeat intervention before, 12 months. RESULTS: Procedures were successful in 23 of 25 patients. There was one immediate occlusion of tibioperoneal arteries, and one early reocclusion of a reopened segment in the ET/PTA group. There was one early reocclusion in the PTA group. At 12 months patency was 70% and 69.2% in the ET/PTA and PTA groups respectively. Covariant analysis showed no significant difference in ABI between the two groups at any of the three measurement times. CONCLUSION: This trial demonstrated no difference between ET/PTA and PTA alone in femoropopliteal occlusions associated with normal proximal arteries and at least one normal tibioperoneal artery. PMID- 9853165 TI - Directional atherectomy in iliac stent failure: clinical technique and histopathologic correlation. AB - PURPOSE: To assess the feasibility and efficacy of directional atherectomy in the treatment of iliac stent stenosis or occlusion and to evaluate the histologic composition of excised atherectomy specimens. METHODS: Directional atherectomy of six occluded and 10 severely stenosed iliac stents was undertaken in 12 patients at a mean interval of 28 months (range 3-69 months) after stent insertion for occlusive aortoiliac disease. In cases of stent occlusion, atherectomy was preceded by low-dose thrombolysis. In all patients stent clearance with return of femoral pulses was achieved within 24 hr and there were no significant complications. All excised specimens were sent for histologic examination. RESULTS: Eleven patients (92%) remain symptom free with unlimited walking distance at a mean follow-up interval of 11.5 months (range 3-31 months) after treatment. Histologic examination revealed typical myointimal hyperplasia at three excision sites, intimal fibrosis at three sites, atheroma at four sites and organized thrombus at six sites. CONCLUSION: Atherectomy offers an effective treatment in iliac stent occlusion and restenosis with no significant adverse effects. Debulking of these lesions seems to offer a more logical approach than simple balloon angioplasty. Clinical and duplex follow-up confirms satisfactory outcome within the first year but longer-term results are not yet known. The histologic data obtained demonstrate that stent restenosis and occlusion are likely to be multifactorial, and challenge the assumption that myointimal hyperplasia is the sole cause of iliac stent occlusion. PMID- 9853166 TI - Can rotational atherectomy cause thermal tissue damage? A study of the potential heating and thermal tissue effects of a rotational atherectomy device. AB - PURPOSE: Thermal tissue damage (TTD) is customarily associated with some lasers. The thermal potential of rotational atherectomy (RA) devices is unknown. We investigated the temperature profile and potential TTD as well as the value of fluid flushing of an RA device. METHODS: We used a high-resolution infrared imaging system that can detect changes as small as 0.1 degree C to measure the temperature changes at the tip of a fast RA device with and without fluid flushing. To assess TTD, segments of porcine aorta were subjected to the rotating tip under controlled conditions, stained by a special histochemical stain (picrisirius red) and examined under normal and polarized light microscopy. RESULTS: There was significant heating of the rotating cam. The mean "peak" temperature rise was 52.8 +/- 16.9 degrees C. This was related to rotational speed; thus the "peak" temperature rise was 88.3 +/- 12.6 degrees C at 80,000 rpm and 17.3 +/- 3.8 degrees C at 20,000 rpm (p < 0.001, t-test). Fluid flushing at 18 ml/min reduced, but did not abolish, heating of the device (11.8 +/- 2.9 degrees C). A crater was observed in all segments exposed to the rotating tip. The following features were most notable: (i) A zone of "thermal" tissue damage extended radially from the crater reaching adventitia in some sections, especially at high speeds. This zone showed markedly reduced or absent birefringence. (ii) Fluid flushing of the catheter reduced the above changes but increased the incidence and extent of dissections in the media, especially when combined with high atherectomy speeds. (iii) These changes were observed in five of six specimens exposed to RA without flushing, but in only one of six with flushing (p < 0.05). (iv) None of the above changes was seen in control segments. CONCLUSION: RA is capable of generating significant heat and potential TTD. Fluid flushing reduced heating and TTD. These findings warrant further studies in vivo, and may influence the design of atherectomy devices. PMID- 9853167 TI - Effect on intimal hyperplasia of dexamethasone released from coated metal stents compared with non-coated stents in canine femoral arteries. AB - PURPOSE: Polymer-coated, dexamethasone (DXM)-releasing stents were tested in order to assess the efficacy of DXM released locally for the prevention of stent restenosis due to intimal hyperplasia. METHODS: Strecker stents coated with a biodegradable membrane containing DXM were implanted percutaneously into the femoral artery in 14 dogs. The contralateral artery received a conventional non coated stent serving as control. The drugs are eluted by degradation of the carrier membrane. Follow-up intraarterial digital subtraction angiography (DSA) was obtained at 3, 6, 9, 12, and 24 weeks with subsequent autopsy. Specimens for gross and microscopic pathology were obtained and histomorphometry was performed. RESULTS: Four of 14 DXM-coated stents showed thrombotic occlusion within the first 3 weeks; ten DXM-coated stents remained patent. At follow-up DSA, DXM coated stents showed a significantly wider lumen than the non-coated stents. At morphometry there was less intimal hyperplasia over DXM-coated stents than over non-coated stents (p < 0.05). CONCLUSION: DXM-coated stents reduce neointimal hyperplasia in dogs when compared with non-coated stents. PMID- 9853168 TI - Polyurethane-coated Dacron-covered stent-grafts for TIPS: results in swine. AB - PURPOSE: To evaluate shunt patency after placing a new polyurethane-coated Dacron covered spiral Z stent-graft to create a transjugular intrahepatic portosystemic shunt (TIPS). METHODS: TIPS stent-grafts were placed in six young swine, using polyurethane-coated, Dacron-covered spiral Z stent-grafts. Animals were followed weekly by transhepatic portal venography until shunt occlusion or up to 6 weeks, then were sacrificed for gross and histologic evaluation. RESULTS: Five of six TIPS stent-grafts were found to be occluded by 3 weeks and none were patent at 6 weeks. Histologic evaluation demonstrated a marked foreign body reaction with superimposed thrombosis occluding the graft lumen. CONCLUSION: Polyurethane coated Dacron-covered stent-grafts do not improve TIPS patency. PMID- 9853169 TI - Treatment of a malignant esophageal perforation with a prototype conical Wallstent. AB - A 60-year-old man with a malignant esophageal perforation could not be treated by conventional covered metallic stents because the upper esophagus was dilated. The perforation was eventually closed by deployment of a prototype, conical covered Wallstent. PMID- 9853170 TI - Transcatheter treatment of life-threatening lower gastrointestinal bleeding due to advanced pelvic malignancy. AB - We present two patients with life-threatening, massive, lower gastrointestinal (GI) bleeding and locally advanced cervical carcinoma. Selective pelvic arteriography demonstrated that the site of bleeding originated from a pseudoaneurysm of the right internal iliac artery with fistulous communication to the sigmoid colon in one patient and from the left internal iliac artery into the rectum in the second patient. Transcatheter embolotherapy was then performed using balloon occlusion in one patient and coil embolization in the second patient. The iliac arteries should also be evaluated in patients with pelvic cancer who present with lower GI bleeding. PMID- 9853171 TI - Treatment of a urinoma and a post-traumatic pseudoaneurysm using selective arterial embolization. AB - We report a case of severe renal trauma giving rise to a pseudoaneurysm of a renal branch artery and a large urinoma secondary to fracture of a calix disconnected from the rest of the collecting system. Both conditions were successfully treated using selective arterial embolization. PMID- 9853172 TI - Use of a puncture needle for recanalization of an occluded right subclavian vein. AB - We report a patient in whom we used a puncture needle to initiate percutaneous recanalization of a chronic occlusion of the junction between the right subclavian vein and the right brachiocephalic vein. Under fluoroscopic guidance, an 18-gauge needle was used to puncture the right subclavian vein. When contrast material injected through the needle confirmed intravascular location, the needle was advanced until it deflected and perforated an occlusion balloon target positioned within the right brachiocephalic vein. This technique may be useful in patients with central venous occlusions that are refractory to traversal using traditional catheter and guidewire techniques. PMID- 9853173 TI - An improved reversal technique from retrograde to antegrade femoral artery cannulation. AB - Converting retrograde femoral access to antegrade access may be required following diagnostic arteriography to perform an interventional procedure. This technical note describes a modification of a previously described technique that allows reliable catheter reversal which was successful in 27 attempted procedures. There were no complications. PMID- 9853174 TI - Re: Friday afternoon pulmonary arteriography (FAPA): replacement by Friday afternoon urokinase (FAUK) PMID- 9853175 TI - [Clinical experience with Danaparoid-sodium: measures to be taken in heparin induced thrombocytopenia]. PMID- 9853176 TI - Sudden cardiac death--can it be defined with precision? PMID- 9853177 TI - Medical and psychological aspects of hormone replacement therapy in postmenopausal women: points of view from a cardiologist and a psychotherapist. PMID- 9853178 TI - Salutary effects of aspirin in coronary artery disease are not limited to its platelet inhibitory effects. AB - Aspirin is widely used in the treatment and prevention of coronary artery disease (CAD). However, other platelet inhibitory agents, which inhibit platelet activation, have not been found to be effective or as effective as aspirin. The discrepancy between the efficacy of these compounds and aspirin suggests that the therapeutic efficacy of aspirin may not be limited to its platelet inhibitory effect. In this review, the basis for a unique place for aspirin in the therapy of patients with CAD is discussed. The author believes that the nonplatelet mediated effects of aspirin could be more important than the platelet inhibitory effect, or at least may complement the platelet inhibitory effects of aspirin in patients with acute myocardial ischemia and in others undergoing intracoronary procedures. PMID- 9853179 TI - Diabetic cardiomyopathy. AB - Prior to 1972, the increased cardiovascular morbidity and mortality that diabetics endure had been attributed to vascular disease. In 1972, Rubler et al. proposed the existence of a diabetic cardiomyopathy based on their expereince with four adult diabetic patients who suffered from congestive heart failure (CHF) in the absence of discernable coronary artery disease, valvular or congenital heart disease, hypertension, or alcoholism. Alternative explanations for CHF, such as anemia and vascular and renal disease in these four patients, gave rise to criticisms, but a wave of subsequent studies in the 1970s and 1980s provided credence to this new disease entity. This review of the studies done since 1972 appears to support the concept of a diabetic cardiomyopathy independent of atherosclerotic cardiovascular disease. The exact mechanism is still questionable, and several mechanisms have been proposed including small and microvascular disease, autonomic dysfunction, metabolic derangements, and interstitial fibrosis. However, the weight of evidence leans toward the development of fibrosis, possibly caused by the accumulation of a peroxidase acid schiff (PAS)-positive glycoprotein, leading to myocardial hypertrophy and diastolic dysfunction. PMID- 9853180 TI - Effects of dichloroacetate in patients with congestive heart failure. AB - BACKGROUND: Conventional approaches to management of congestive heart failure (CHF) rely on drugs that increase myocardial contractility or reduce ventricular afterload. These approaches often improve cardiac symptoms and survival, but may be associated with significant deleterious effects. An alternative approach is to enhance myocardial energy production. Dichloroacetate (DCA) stimulates pyruvate dehydrogenase activity and accelerates aerobic glucose, pyruvate, and lactate metabolism in myocardial cells. These alterations would be expected to improve myocardial function. HYPOTHESIS: The purpose of the investigation was to assess the efficacy of DCA in patients with left ventricular systolic dysfunction and to examine the mechanism by which improvement occurs. METHODS: A total of 25 patients (16 men, 9 women; age range 31-72 years, mean 59) with CHF and ejection fraction < or = 40% received an intravenous infusion of 50 mg/kg DCA over 15 min. Indices of systolic and diastolic function were obtained by two-dimensional and Doppler echocardiography performed at baseline, 30 min, and 60 min following completion of DCA infusion. RESULTS: Baseline ventricular ejection fraction was 27.3 +/- 9.1%; 17 patients (68%) had nonischemic cardiomyopathy. Heart rate increased after DCA infusion from 73.9 +/- 14.5 to 79.2 +/- 14.9 beats/min at 60 min; p = 0.02. Left ventricular diastolic and systolic volumes increased at 30 min compared with baseline (248.7 +/- 98.1 vs. 259.6 +/- 99.6; p = 0.04, and 180.1 +/- 80.4 vs. 192.2 +/- 84.9; p = 0.002, respectively), but stroke volume (49.2 +/- 19.1 vs. 48.9 +/- 18.1; p = 0.9) and ejection fraction (27.3 +/- 9.1 vs. 25.7 +/- 9.8; p = 0.2) were unchanged. Indices of diastolic function were also unchanged. CONCLUSION: Dichloroacetate infusion in patients with CHF is not associated with improvement in noninvasively assessed left ventricular function. PMID- 9853181 TI - The effects of an angiotensin-converting inhibitor (enalapril) on patients with mild cardiac failure--evaluating cardiac function based on the relationship between daily walking pace and heart rate. AB - BACKGROUND: Heart failure has been evaluated by several methods, the New York Heart Association (NYHA) classification of heart failure based on symptoms being used most frequently. However, the degree of heart failure assessed by these criteria does not always correlate with cardiac function in daily life. HYPOTHESIS: The aim of the study was to evaluate cardiac function based on the walking pace/heart rate (HR) relationship to assess the effects of enalapril, an angiotensin-converting enzyme inhibitor, in patients with mild to moderate cardiac function. METHODS: To evaluate cardiac function objectively, we developed a method using a pedometer to count the steps walked while simultaneously recording HR using a Holter electrocardiograph (ECG). Step-count walk rate (WR) was recorded on the magnetic tape of the Holter apparatus, and both HR and walking pace were calculated automatically by the Holter ECG analysis system. Data were determined every hour, and mean pace and HR were plotted along the x and y axes, respectively. The slope of HR x WR was calculated using the least squares method. The slope and the total number of steps were regarded as indicators of cardiac function and quality of life, respectively. We analyzed 36 subjects, consisting of 8 normal volunteers, 8 patients in New York Heart Association (NYHA) class I. 11 in class II, and 9 in class III chronic mild heart failure, during maximal exercise work load by bicycle ergometer; furthermore, fractional shortening of the left ventricle on echocardiogram was determined in 14 patients with chronic mild heart failure and was compared with the slope of HR x WR. Enalapril was administered at a daily dose of 2.5-10 mg for 1-24 months (mean 6 months) in 60 patients to evaluate the effects of this drug on these parameters. RESULTS: There was a significant inverse relationship between maximal work load and the HR x WR slope, and also between the fractional shortening and the slope, suggesting that the slope may reflect the severity of cardiac dysfunction. Furthermore, the slope decreased significantly from 1.8 +/- 1.26 before enalapril to 1.0 +/- 0.94 (mean +/- standard deviation) after drug administration, while the total number of steps increased significantly from 4842 +/- 3581 to 7804 +/- 4793. CONCLUSION: The slope of the graph relating step count and HR proved to be a good, objective indicator of cardiac function, and enalapril therapy improved this parameter. PMID- 9853182 TI - The impact of beta-receptor blocker addition to high-dose angiotensin-converting enzyme inhibitor-nitrate therapy in heart failure. AB - BACKGROUND: The natural history of heart failure is one of continued worsening of cardiac function. Beta-receptor blocker therapy has been effective in improving clinical status and left ventricular function in patients with heart failure. Similarly, high doses of angiotensin-converting enzyme (ACE) inhibitors with nitrates partially reverse the ventricular remodeling of heart failure. HYPOTHESIS: We tested the hypothesis that beta-blocker therapy added to high-dose ACE inhibitor-nitrates would potentiate the reversal of heart failure. METHODS: Thirteen patients, aged 52 +/- 8 years, with moderate to severe heart failure, 12 of whom were referred for transplant consideration, with heart failure duration of 4.8 +/- 2.2 years, were prospectively followed for 12 months. Baseline echocardiographic ejection fraction was 19 +/- 8%, and presenting New York Heart Association class was 2.9 +/- 0.7. Angiotensin-converting enzyme inhibitors and nitrates were uptitrated over 6 months to a final dose of lisinopril 53 +/- 31 mg/day, and isosorbide dinitrate 217 +/- 213 mg/day. At 6 months, beta-blocker therapy with atenolol was initiated and titrated to a final dose of 46 +/- 23 mg/day. Two-dimensional Doppler echocardiography and metabolic stress testing were performed at baseline, at 6 months on lisinopril-nitrates only, and at 12 months on combined ACE inhibitor-nitrate and beta-blocker therapy. RESULTS: New York Heart Association classification improved from 2.9 +/- 0.7 to 1.8 +/- 0.8 on lisinopril-nitrates (p < 0.05), and to 1.5 +/- 0.5 with the addition of beta blockade (p = NS). On follow-up, peak oxygen consumption rose from 17 +/- 7 ml O2/kg/min at baseline to 21 +/- 5 ml O2/kg/min at 6 months on lisinopril-nitrates (p = 0.06) without further change on beta blockade. Ejection fraction rose from 19 +/- 8 to 33 +/- 14% on lisinopril-nitrates at 6 months (p = 0.005) and to 36 +/- 18% on beta blockade at 12 months (p = NS). CONCLUSION: High-dose ACE inhibitor-nitrate therapy significantly improved patient clinical status and left ventricular systolic function in heart failure. The addition of beta-receptor blockade over and above high-dose ACE inhibitor-nitrates was well tolerated but had no further impact on symptomatic status, exercise tolerance, or left ventricular systolic function. The potential for pharmacologic reversal of heart failure remodeling may be finite despite the use of complementary therapies. Larger placebo-controlled and randomized trials of beta-receptor blockade added to high-dose ACE inhibitor-nitrate therapy are needed to confirm these observations. PMID- 9853183 TI - Prognostic implications of ventricular ectopy one week, one month, and sixteen months after an acute myocardial infarction. Danish Study Group on Verapamil in Myocardial Infarction. AB - BACKGROUND: Ventricular ectopy early after an acute myocardial infarction (AMI) has previously been demonstrated to predict mortality. Less information is available about the prognostic implications of ventricular ectopy occurring late after an AMI, and no information is available about the prognostic implication of the development of ventricular ectopy during the first year after an AMI. HYPOTHESIS: The purpose of the present prospectively conducted trial, a part of the Danish Verapamil Infarction Trial II (DAVIT II), was to evaluate the prognostic implication of (1) ventricular premature complexes (VPCs) recorded by 24-h Holter monitoring 1 week, 1 month, and 16 months after an AMI; and (2) development of > 10 VPCs/h or of any complex ventricular ectopy, that is, pairs, more than two types of VPCs, ventricular tachycardia, or > 10 VPCs/h during follow-up after an AMI. METHODS: Patients were monitored 1 week (n = 250), 1 month (n = 210), and 16 months (n = 201) after AMI. RESULTS: Multivariate analyses based on history, clinical findings, and ventricular ectopy showed the following results: After 1 week, > 10 VPCs/h (p = 0.0006) and heart failure (p < 0.007); after 1 month, > 10 VPCs/h (p = 0.003) and resting heart rate (p < 0.02); and after 16 months, ventricular tachycardia (p = 0.002) independently predicted long-term mortality. Mortality was significantly predicted by the development of > 10 VPCs/h from 1 week to 1 month (p = 0.003) and 16 months (p = 0.03), and from 1 to 16 months (p = 0.007) after AMI, as well as by the development of any complex ventricular ectopy from 1 week to 1 month (p = 0.02) and 16 months (p = 0.01), and from 1 to 16 months (p = 0.04) after AMI. CONCLUSION: The present study demonstrated that 1 week and 1 month after an AMI the quantity of VPCs, that is, > 10 VPCs/h, predicted mortality, whereas 16 months after an AMI the quality of VPCs, that is, ventricular tachycardia, predicted mortality. PMID- 9853184 TI - Characteristics of patients less than 45 years of age compared with older patients undergoing coronary artery bypass grafting. AB - BACKGROUND: Coronary artery disease (CAD) was not recognized as common among young patients until the study by Yater in 1948. Subsequent studies further elucidated the nature of the disease, which had become more apparent in the younger groups. HYPOTHESIS: The study was undertaken to determine the prevalence of risk factors and severity of coronary disease among young patients aged < or = 45 years undergoing coronary artery bypass grafting (CABG) compared with older patients. METHODS: In all, 112 young patients aged < or = 45 years (Group 1) and 798 older patients aged > 45 years (Group 2) were analyzed for trends to hypertension, smoking, diabetes, family history of heart disease, hypercholesterolemia, obesity, and history of previous myocardial infarction (MI). The severity of disease was examined in terms of number of diseased vessels, vessel size, number of grafts performed, performance of endarterectomies, and left ventricular function. RESULTS: Group 1 had a higher incidence of positive family history (68.5 vs. 51.2%, p < 0.05), and lower incidences of hypertension (62.7 vs. 81.5%, p < 0.05), obesity (42.9 vs. 83.9%, p < 0.05), and history of previous MI (54.5 vs. 94.6%, p < 0.05). Group 2 had a higher incidence of left main disease (22.6 vs. 11.4%, p < 0.05). The distribution of the affected vessels of the young patients was most commonly the left anterior descending (90.4%) followed by the right coronary (79.8%) and circumflex arteries (69.2%). Group 2 had more grafts per patient (3.82 vs. 3.37, p < 0.05). The size of the diseased vessels measured intraoperatively was similar (1.56 vs. 1.58 mm, p = NS) in both groups. Endarterectomy was performed almost three times more often in Group 1 patients (8.2 vs. 3.0%, p < 0.05). Operative mortality was less in Group 1 mean (1.8 vs. 6.3%, p < 0.05). Group 2 had a greater mean left ventricular ejection fraction (53.8 vs. 49.7%, p < 0.05). CONCLUSION: Compared with the older population, patients < or = 45 years of age who underwent CABG had (1) a higher incidence of positive family history of CAD, (2) a higher likelihood of requiring an endarterectomy, and (3) lower operative mortality rate despite a slightly poorer ventricular function. PMID- 9853185 TI - Left ventricular aneurysm after myocardial infarction. PMID- 9853186 TI - An unusual cardiomegaly: the giant left atrium. PMID- 9853187 TI - Endovascular stenting of an unprotected left main coronary artery stenosis in a heart transplant patient. AB - Endoluminal revascularizaion of left main coronary artery vessels is considered to be relatively contraindicated because of a high procedural mortality and restenosis rate. This report describes the first successful case of endovascular stenting in an unprotected left main coronary artery stenosis in a heart transplant patient. PMID- 9853188 TI - Noble O. Flower: the complete cardiologist. PMID- 9853189 TI - The role of third-generation beta-blocking agents in chronic heart failure. AB - Third-generation beta-blocking agents developed for the hypertension market are proving useful in the treatment of chronic heart failure (HF). These compounds share the ancillary property of vasodilation, which improves acute tolerability by unloading the failing left ventricle at a time when beta-adrenergic withdrawal produces myocardial depression. In the case of carvedilol and bucindolol, this allows for the administration of nonselective beta blockade. Because of blockade of both beta 1 and beta 2 adrenergic receptors as well as other properties, these compounds possess a more comprehensive antiadrenergic profile than second generation, beta 1-selective compounds. For this and potentially other reasons, third-generation beta-blocking agents have theoretical efficacy advantages that have yet to be demonstrated in large-scale trials. Ongoing trials with either second- or third-generation compounds and one trial directly comparing a compound from each class will provide the answer as to whether third-generation compounds have an advantage in the treatment of chronic HF. PMID- 9853190 TI - Cardiac remodeling as a consequence and cause of progressive heart failure. AB - Natural history studies in heart failure have shown that increases in left ventricular (LV) volume and LV mass are directly related to future deterioration in LV performance and a less favorable clinical course. Despite the recognized importance of remodeling in heart failure, very little is known about the basic mechanisms that lead to cardiac remodeling. In this review, we will summarize recent clinical and experimental studies that highlight the importance of the remodeling process during the progression of heart failure. The intent of this review is to provide an integrated view of the mechanisms that contribute to LV remodeling at the cellular level, the myocardial level, and the level of the chamber. PMID- 9853191 TI - The effects of norepinephrine on myocardial biology: implications for the therapy of heart failure. AB - Increased sympathetic nervous system (SNS) activity in patients with heart failure may help to support cardiovascular function. However, increased SNS activity, particularly if prolonged, may exert deleterious effects on cardiovascular structure and function by stimulating pathologic myocardial remodeling. In vitro, norepinephrine mimics many features of myocardial remodeling, including hypertrophy of individual myocytes and reinduction of fetal genes. Furthermore, stimulation of the beta-adrenergic pathway has been shown to stimulate apoptosis of cardiac myocytes in vitro, in rats infused with isoproterenol, and in mice that overexpress the stimulatory G-protein, Gs. Thus, increased SNS activity, acting via beta-adrenergic receptors, may play an important role in the progression of myocardial failure by acting directly on myocytes and other cell types in the heart to regulate fundamental biologic properties such as growth, apoptosis, and the composition of the extracellular matrix. This thesis provides a mechanism by which beta-adrenergic antagonists may inhibit or reverse pathologic remodeling, improve myocardial structure and function, and prolong patient survival. PMID- 9853192 TI - Significance of reduction in heart rate in cardiovascular disease. AB - Epidemiologic studies suggest that lower heart rate is associated with decreased cardiovascular and all-cause mortality. Heart rate has also been reported to be an independent predictor of outcome after myocardial infarction (MI). Because it is a major determinant of oxygen consumption and metabolic demand, a decrease in heart rate would be expected to decrease cardiac workload. Among patients with restricted coronary blood flow, increased heart rate is associated with more severe myocardial ischemia, angina, and an increase in size of MI. Pharmacologic interventions that reduce heart rate, such as beta blockers, generally reduce mortality and improve outcome. A number of clinical trials using beta blockers after MI has shown a relationship between reduction in heart rate and reduction in mortality. Most beta blockers demonstrate a nearly linear relationship between reductions in mortality and in heart rate. The evidence from trials with calcium antagonists is more equivocal, possibly because until very recently none were available that decreased heart rate without decreasing myocardial contractility. Drugs that do not reduce the heart rate after an MI and in congestive heart failure have not been found to improve survival. In light of the findings thus far reported, reduction in heart rate should be a therapeutic goal in the treatment of cardiovascular disease. PMID- 9853193 TI - Why we should attempt to eliminate (silent and symptomatic) ischemia in patients with chronic stable angina. AB - There is no doubt that angina patients with symptomatic ischemia face a poor prognosis and that treatment of these patients is necessary. However, there is no consensus regarding the prognostic value or treatment of silent ischemia in patients with stable angina. It is in this context that the advantages of treating silent ischemia are presented. PMID- 9853194 TI - The safety of calcium-channel blockers. AB - Calcium-channel blockers are widely used as an effective treatment for hypertension and angina. Several studies have raised questions about their safety, suggesting that calcium-channel blockers can increase the rates of myocardial infarction (MI) and death, particularly in patients with heart disease. Reviews of these studies have uncovered serious methodological shortcomings or have found them restricted to short-acting drugs, frequently at high doses or used inappropriately. One study was based on old data regarding only short-acting nifedipine, which has never been indicated for patients who have suffered an MI or unstable angina. A case-control study of short-acting verapamil, diltiazem, and nifedipine suggested an increased MI rate was confounded by the higher rates of diabetes and preexisting heart disease in the patients treated with calcium-channel blockers. A third study reported significantly decreased survival only in patients taking short-acting nifedipine; in most of the cases reported, blood pressure was not controlled. While these studies alert us to the limitations of short-acting calcium-channel blockers and the necessity of considering side effects such as neurohormonal stimulation, a number of more recent, better-controlled studies have not confirmed increased risk with calcium-channel blockers when appropriately employed. Calcium-channel blockers should still be considered first-line therapy in appropriately selected patients with hypertension or angina. PMID- 9853195 TI - Is coronary revascularization the optimum strategy for patients with non-Q wave myocardial infarction? A point-counterpoint. PMID- 9853196 TI - Update on diagnostic methods, natural history and outcome variables in Alzheimer's disease. AB - The diagnosis of Alzheimer's disease (AD) currently relies on history obtained from family or friends and on mental status assessment matched to National Institute of Neurological and Communicative Disorders and Stroke criteria. Progression over time may or may not be typical, suggesting alternate diagnoses such as Lewy body or frontotemporal dementias. Apolipoprotein E genotype does not appear to be useful as a diagnostic marker. The usefulness of brain imaging in AD must be reexamined. Critical events in the natural history of AD, such as institutionalization and loss of ability for self-care, could be used as end points. Loss of ability for instrumental tasks, such as driving, traveling alone, or managing finances, would be preferable for early-stage stabilization studies. Different symptomatic domains of AD (mood, cognition, functional autonomy, behavior, motoricity) can be quantified using specific outcome measures. Although cognitive loss has been considered a core symptom of AD from a regulatory perspective, loss of functional autonomy and behavioral disinhibition are considered more important by clinicians and families. Recently, the availability of new scales has led to an interest in all of these domains. Results from symptomatic drug studies suggest a differential effect of cholinesterase inhibitors on cognition versus muscarinic agonists on functional autonomy and behavior. Hence there is a need to measure these domains separately and, eventually, to attempt combination therapy. Quality of life is a difficult but important dimension of AD therapeutic research, and it requires further methodological research. PMID- 9853197 TI - Global change assessments in anti-Alzheimer clinical drug trials. AB - The assessment of efficacy in clinical trials in Alzheimer's disease has been a controversial area. The US Food and Drug Administration has not accepted mental status assessment as a sole determinant of efficacy because of concerns about oversensitivity and lack of direct linkage to clinical importance. Global change instruments offer a number of advantages as primary outcome measures, chief among which is a wide range of responsiveness, a property well suited for trials of 1 to 2 years. Initial investigations with global change instruments have shown encouraging validity. Reliability has been only moderate, suggesting that the methodology needs further attention. A standardized format for interview and the acceptance of a set of anchor points that better define the ratings of change are two modifications that should improve reliability. Global change instruments, if properly explained to the patients, caregivers and primary physicians, will capture the concept of clinical importance. PMID- 9853198 TI - Clinical subtypes of Alzheimer's disease. AB - Alzheimer's disease (AD) can present as a variety of clinical profiles. Although the most common presentation is that of a progressive amnestic disorder with subsequent involvement of other cognitive functions and personality alterations, there are numerous other clinical profiles. AD can present as a focal cortical degenerative syndrome with the clinical features dependent on the regions of the brain involved. Some of these syndromes include disturbances of language, visuospatial skills, attentional functions, executive processes and praxis. The neuropathological substrate of these disorders is variable and can include AD. Recently, the Lewy body variant of AD has been described. Finally, other modifying features that affect the progression of AD, such as extrapyramidal symptoms and myoclonus, are also discussed. Although the progressive amnestic form of AD is the most common presentation, other variations on the clinical syndrome can be important to identify because they may have implications for prognosis and treatment. PMID- 9853199 TI - The caregiver's role in Alzheimer's disease. AB - The majority of persons with Alzheimer's disease (AD) are cared for by family members in their home. Research from several studies has shown that these care providers have rates of emotional morbidity three times that seen in age-similar individuals. This paper will review the epidemiological data on the incidence and prevalence of emotional disability in caregivers. It will also review the 11 treatment studies in the literature in which blinded intervention was carried out. Nine of these 11 studies show that intervention was more beneficial than the placebo comparison treatment. Both educational and emotional support were effective. However, when compared with each other, emotional support was more effective; the combination may be more effective than either alone. Preliminary evidence suggests that intervention on behalf of caretakers may delay the need for institutionalization of family members with AD. PMID- 9853200 TI - Perspectives in the management of Alzheimer's disease: clinical profile of donepezil. AB - Donepezil HCI is a piperidine-based reversible acetylcholinesterase (AChE) inhibitor, chemically distinct from other cholinesterase (ChE) inhibitors and rationally designed to treat the symptoms of Alzheimer's disease (AD). It is highly selective for AChE in the central nervous system (CNS), with little or no affinity for butyrylcholinesterase (BuChE). In preclinical studies in animals, donepezil produced increased CNS acetylcholine. The resultant enhancement of cholinergic activity gave rise to improved performance by rats on tests of learning and memory, with no evidence of hepatic or renal toxicity. In subsequent phase I clinical evaluations in healthy volunteers, donepezil demonstrated favorable pharmacokinetic, pharmacodynamic and safety profiles. Its long terminal disposition half-life supported once-daily administration, with no requirement for dose modification in the elderly or in patients with renal or hepatic impairment. A 14-week, phase II dose-finding study in patients with mild to moderate AD (Clinical Dementia Rating [CDR], 1-2; Mini-Mental State Examination [MMSE], 10-26) showed that donepezil at a dose of 5 mg/day produced highly significant improvements in cognition (as measured by the Alzheimer's Disease Assessment Scale, cognitive subscale [ADAS-cog]). Subsequently, two pivotal parallel-group, placebo-controlled phase III trials (of 15 and 30 weeks' duration) showed highly statistically significant improvements in ADAS-cog, MMSE, Clinician's Interview-Based Impression of Change with caregiver input (CIBIC plus) and CDR-SB (Sum of the Boxes) scores, compared with placebo, in mild to moderate AD patients treated with either 5 or 10 mg/day donepezil. Adverse events in the phase II and III trials were mild and transient and resolved with continued donepezil administration. The donepezil clinical trials program has shown that this drug is a clinically effective and well-tolerated once-daily treatment for the symptoms of mild to moderate AD. PMID- 9853201 TI - Efficacy and acceptability of two monophasic oral contraceptives containing ethinylestradiol and either desogestrel or gestodene. AB - OBJECTIVE: To assess the contraceptive efficacy, cycle control and acceptability of two monophasic oral contraceptives containing either 30 micrograms ethinylestradiol plus 150 micrograms desogestrel or 30 micrograms ethinylestradiol plus 75 micrograms gestodene. METHODS: In a randomized, open label, six-cycle, group-comparative, multicenter study performed in Brazil, pregnancies, cycle-control parameters, incidence of side-effects and the presence and severity of acne vulgaris were assessed, and blood pressure and body weight were measured at pretreatment and after one, three and six cycles of oral contraceptive use. RESULTS: Of the 595 women enrolled, 274 (86.7%) in the desogestrel/ethinylestradiol group and 227 (81.4%) in the gestodene/ethinylestradiol group completed the six cycles, providing data for 1753 and 1487 treatment cycles, respectively. Two pregnancies occurred, one of which (in the desogestrel/ethinylestradiol group) was attributed to user failure, whilst the other (in the gestodene/ethinylestradiol group) was thought to result from method failure. Cycle control was observed to be excellent; the incidences of irregular bleeding and minor side-effects were low in both groups and decreased after an initial increase in the first cycle. Pre-existing acne improved in both groups, whereas blood pressure and body weight remained essentially unchanged. CONCLUSIONS: Both desogestrel/ethinylestradiol and gestodene/ethinylestradiol provide effective oral contraception with comparable cycle control and acceptability. PMID- 9853203 TI - Increasing the effectiveness of contraceptive usage in university students. AB - Prevention of pregnancy requires correct and consistent use of an effective method of contraception and knowledge alone is not sufficient to ensure such use, as many complex social and behavioral factors influence contraceptive behavior. Women are particularly likely to change their contraceptive method after a contraceptive 'shock'. In this study, the change in contraceptive behavior of a group of university students who presented for emergency contraception is studied. Each student participated in a single individualized educational session. A total of 465 women requested emergency contraception in a 3.5-year period at a large student health center. Of these, 24% had not previously used contraception, 50% had previously used condoms and 25% had taken the pill (COC). Of those who usually used condoms, 79% had had a condom accident and 21% had not used them at the last intercourse. Only 30% of COC users had had a problem with the pill and the remaining 70% were not taking it at the time of last intercourse, for social rather than medical reasons. Follow-up is available for 309 (66.5%). Paired-sample analysis of these women shows a decrease in the number using no contraception (22% to 8%), and an increase in the number using COCs (33% to 66%). These changes reached statistical significance (p < 0.0001). University students, despite their intelligence, exhibit a high degree of risk-taking behavior but become more effective contraceptive users after an interactive counselling session following a contraceptive scare. PMID- 9853204 TI - Inhibins, activins and follistatins: a review of complex regulators of the reproductive system. PMID- 9853202 TI - Flexible starting schedule for oral contraception: effect on the incidence of breakthrough bleeding and compliance. AB - OBJECTIVE: To compare the effect of starting oral contraceptives on the first day of menses with the effect of starting on the day of menses' cessation (but no later than the 5th day following its onset), on the incidence of early breakthrough bleeding. METHOD: Oral contraceptives containing 30 micrograms ethinylestradiol and 75 micrograms gestodene were prescribed to 200 consecutive healthy women in whom oral contraceptives were found to be the most suitable method of contraception. In the first 100 women, treatment was started on the 1st day after the onset of menses (Day 1 group), and in the remainder, treatment was started on the day of menses' cessation, but no later than the 5th day following its onset (Flexible group). RESULTS: The Flexible group had better compliance and a reduced incidence of breakthrough bleeding. No differences were observed between the two groups for age, parity and gravity, or contraceptive failure. CONCLUSIONS: Oral contraceptives may be initiated on the day of menses' cessation, but no later than the 5th day following its onset. This regimen might increase patient compliance and lower the incidence of breakthrough bleeding, probably without adversely affecting contraceptive efficacy. PMID- 9853205 TI - Improvement of surveillance programs for sexually transmitted infections and related conditions. AB - The present communication considers possibilities of how to improve surveillance programs for sexually transmitted infections and some conditions not yet included among the 'established' sexually transmitted infections. An extended list of sexually transmitted infection agents is proposed for monitoring and a syndrome based surveillance program for what and when to screen pregnant women is discussed. Problems of how to deal with recurrences in chronic sexually transmitted infections are discussed. The value of concomitant surveillance programs, for clinicians and laboratories, is stressed. The importance of reporting the method(s) used to establish the diagnosis, as well as the circumstances under which the sampling was made, for example at screening, is also highlighted. PMID- 9853206 TI - Limitations of current surveillance programs for sexually transmitted infections and allied conditions affecting reproductive health care. AB - The present communication concerns the potential of current surveillance programs for understanding the epidemiology of sexually transmitted infections. The often limited knowledge or the negligence of such infections both among health providers and in the general population, and the non-existence of laboratory services or poor utilization thereof, for example the lack of sampling material, lack of transport services for samples, lack of laboratory reagents and the use of unsensitive and unspecific detection methods, challenge the value of almost any surveillance system of these infections. Other factors are poor health seeking behavior in the population, including hard-core groups for sexually transmitted infections, poor health economy, limited possibilities for attendees themselves to pay for etiological tests and lack of educated health providers for this type of infectious disease. Still other factors adding to the difficulty of interpreting sexually transmitted infection epidemics on the basis of monitored data are, for example, anonymous testing, double or multiple registration of detected sexually transmitted infection cases, and the chronicity of many sexually transmitted infections with relapses, which makes it difficult to differentiate between incidence and prevalence. Furthermore, partner notification legislations and bureaucracy in reporting cases can influence surveillance programs for sexually transmitted infections. PMID- 9853207 TI - Legal abortion in Denmark during the past 25 years: aspects of public health and ethics. AB - Since October 1, 1973, Denmark has granted its permanent residents the right to legal abortion up to the end of the 12th week of gestation. In the beginning of the period of legal abortion, the numbers of induced abortions per year were high, although they decreased during the 1980s and 1990s. Probably, information campaigns concerning the use of contraception have had some effect. Abortion figures are, however, of interest of studied as rates of abortion, i.e. numbers per 1000 women of fertile age or as age-related rates of abortion. Aspects of legal abortion comprise various topics. One of these is the ethical questions for physicians and nurses, and also for the entire population in a country with legal, and free, abortion to a certain limit. Some central ethical questions are discussed, and it is stated that we cannot perform abortion if we grant the fetus the same moral status as an adult human being. PMID- 9853208 TI - The FemCap: a new contraceptive choice. AB - OBJECTIVE: To develop a female barrier to, first, prevent pregnancy and, second, protect against sexually transmitted diseases. DESIGN: The contraceptive is made of inert, soft material and is an anatomical design that utilizes the physiology of the vagina. It incorporates the use of microbicidal spermicide to kill sexually transmitted disease organisms. RESULTS: FemCap has been proven in clinical trials to be safe, and has no systemic or local side-effects, and does not increase the risk of urinary tract infections. Effectiveness in pregnancy prevention in 86.5% in typical use and 98% if used properly with emergency contraceptives as back-up. FemCap is highly acceptable to women and to their partners. CONCLUSIONS: A new method of birth control has arrived that is safe, effective, highly acceptable and may protect against sexually transmitted diseases of the upper female genital tract. PMID- 9853209 TI - Magnetic resonance imaging of normal bone marrow. AB - The appearance in magnetic resonance imaging (MRI) of the bones depends, to a large extent, on the unmineralized content of the bone cavities. Because yellow marrow contains a large number of fat protons and red marrow a significant number of water protons, MRI offers the opportunity to map the distribution of red and yellow marrow. In addition, red marrow MR appearance varies according to the relative proportion of fat and nonfat cells. Variations in the composition of red marrow and its distribution among normal subjects, mainly in relation to age and sex, contribute to creating a wide spectrum in bone MR appearance, which must be known in order to avoid confusion with bone marrow abnormalities. PMID- 9853210 TI - Magnetic resonance imaging of the bone marrow in hematological malignancies. AB - Despite its lack of specificity, magnetic resonance imaging (MRI) of the bone marrow has the potential to play a role in the management of patients with primary neoplastic disorders of the hematopoietic system, including lymphomas, leukemias and multiple myeloma. In addition to its use in the assessment of suspected spinal cord compression, bone marrow MRI could be used as a prognostic method or as a technique to assess the response to treatment. The current review addresses the common patterns of bone marrow involvement observed in primary neoplasms of the bone marrow, basic technical principles of bone marrow MRI, and several applications of MRI in selected clinical situations. PMID- 9853211 TI - MRI of bone metastases. AB - Some knowledge of MR theory is required to be able to achieve high contrast between bone metastases and normal marrow. Three factors are used in MR to diagnose bone metastases; fat-water distribution, artifacts induced by bone trabeculae, and uptake of contrast medium. Using MR-histological correlations based on specimens of the lumbar spine, and studies of patients, we explain the advantages and limitations of sequences studying fat and water (spin-echo T1, STIR, in- and out-of-phase gradient echo, fat presaturation), bone trabeculae (gradient echo with long TE), and the injection of contrast medium. PMID- 9853212 TI - Amyloid arthropathy of the hip joint: MR demonstration of presumed amyloid lesions in 152 patients with long-term hemodialysis. AB - The aim of this study was to determine the spectrum of MR findings of presumed amyloid arthropathy of the hip joints in patients on long-term hemodialysis. We prospectively performed T1- and T2-weighted spin-echo imaging on 152 consecutive patients on hemodialysis. The duration of hemodialysis ranged from 5 months to 24 years, 2 months (mean: 8 years, 8 months). The frequency, location, and signal intensity of bone lesions were assessed. In 12 cases with contrast-enhanced MR examination, enhancement pattern of bone lesions, synovial lesions, and intra articular lesions were characterized. Bone lesions presumed to be amyloid deposits were identified in 60 patients (39%). Magnetic resonance imaging revealed that amyloid lesions were more extensive than anticipated by plain radiographs. All bone lesions showed decreased signal intensity on T1-weighted images. On T2-weighted images, bone lesions showed increased signal intensity in 32 patients (54%), decreased signal intensity in 11 patients (18%), and both increased and decreased signal intensity in 17 patients (28%). Following intravenous injection of gadolinium-based contrast, all bone lesions showed moderate enhancement. Synovial thickening could not be identified on T1- and T2 weighted images. However, contrast-enhanced images showed thickened synovial membrane, which could be differentiated from joint fluid. Intra-articular nodules showed decreased or intermediate signal intensity on T1-weighted images and decreased signal intensity on T2-weighted images; the intra-articular nodules were contiguous with subchondral bone lesions. Magnetic resonance imaging is useful for evaluating the distribution and extent of amyloidosis of the hip joints in patients undergoing long-term hemodialysis. PMID- 9853213 TI - Langerhans cell histiocytosis of the clavicle: a case report. AB - A case of Langerhans cell histiocytosis in a 47-year-old male presenting as an aggressive appearing lesion of the clavicle is reported. It illustrates the difficulties of the radiological diagnosis of a solitary bone lesion. PMID- 9853214 TI - MR imaging of a malignant schwannoma and an osteoblastoma with fluid-fluid levels. Report of two new cases. AB - One case of malignant schwannoma of the sacrum and another of occipital osteoblastoma were evaluated by MR imaging. Both tumors showed fluid-fluid levels with different signal intensities in the sequences performed. Pathologic examination revealed hemorrhagic fluid in both tumors. Malignant schwannoma and osteoblastoma should be included in the list of bone and soft-tissue with fluid fluid levels. Our data confirm the non-specificity of this finding, which only suggests the presence of previous intratumoral hemorrhage. PMID- 9853215 TI - Subcutaneous granuloma annulare: MR imaging and literature review. AB - Subcutaneous granuloma annulare (SGA) is little known to radiologists. Better knowledge of this lesion may prompt accurate diagnosis. A typical case is presented with plain radiography, ultrasound and MR imaging, and is confirmed by histology. When an otherwise healthy child presents with a rapidly growing, solitary, nontender, subcutaneous soft tissue mass, located on the scalp or extensor aspect of the limbs, that radiologically presents as an indistinct radiodense and hypoechoic mass, isointense to muscle on T1- and slightly hypointense to fat on T2-weighted MR images, without calcifications, bone involvement or extracompartmental invasion, SGA should be suspected. PMID- 9853216 TI - Bone and skeletal muscle metastases from gastric adenocarcinoma: unusual radiographic, CT and scintigraphic features. AB - Skeletal metastatic lesions arising from gastric cancer are uncommon and usually of the osteolytic type. In order to gain a better understanding of its radiological spectrum, we report two atypical cases of skeletal metastases from gastric adenocarcinoma presenting with unusual radiographic, CT and scintigraphic features. In one patient multiple ossifying skeletal muscle metastases and bone metastases with spiculated periosteal reaction occurred as a presenting manifestation of the malignant disease. The other patient developed widespread osteosclerotic metastases with a superscan pattern on bone scintigraphy. PMID- 9853217 TI - High-resolution magnetic resonance angiography of the internal carotid artery: 2D vs 3D TOF in stenotic disease. AB - The aim of this study was to compare high-resolution 2D TOF with high-resolution 3D TOF in the study of internal carotid artery disease. Sixty-four patients with clinical signs of cerebrovascular insufficiency were studied with a superconductive 1.5 T magnet using two techniques: 2D and 3D TOF. Digital subtraction angiography (DSA) was the gold standard. The 2D TOF technique was performed using the following parameters: TR/TE/FA/MA 49 ms/9 ms/60 degrees/512 x 256; the 3D TOF was performed with the following parameters: TR/TE/FA/MA 50 ms/8 ms/20 degrees/512 x 256. The 2D TOF agreed with DSA in 116 of 128 diagnostic judgments (90%) and overestimated seven times. The 3D TOF technique agreed with DSA in 125 of 128 diagnostic judgments (97%) with one overestimation and two underestimations. There was no statistically significant difference (P < 0.05) between the two different techniques. Our study confirms the high reliability of the methodology carried out with the high-resolution 2D and 3D technique. PMID- 9853218 TI - Aortitis with antiphospholipid antibodies: CT and MR findings. AB - Two cases of aortitis associated with the presence of antiphospholipid antibodies (APAs) are reported. Only CT and MR imaging were able to show these unusual form of aortitis preferentially affecting the outer aortic tunics. We conclude that aortitis could be a new manifestation of primary antiphospholipid syndrome (APS) and the initial pathological process before the development of aortic thrombosis, reported as a classical complication of APS. PMID- 9853219 TI - Spiral CT of pulmonary embolism: diagnostic approach, interpretive pitfalls and current indications. AB - The introduction of spiral CT has recently modified the diagnostic work-up of pulmonary embolism, because it is possible to depict noninvasively endoluminal clots in second-to-fourth-division pulmonary arteries. If this technique is currently considered a powerful imaging alternative for the detection of acute central emboli, it is mainly related to the possibility to obtain a uniform and high degree of arterial enhancement of pulmonary arteries down to 2-3 mm in diameter. Minimal experience in spiral CT angiography is necessary to achieve this goal and requires familiarity with both data acquisition and contrast medium injection. Numerous interpretive pitfalls exist in assessing spiral CT images, and certain caveats have to be heeded. However, their recognition becomes increasingly less problematic as the radiologist gains experience with spiral CT of the pulmonary vasculature. Therefore, the purpose of this article is to review the diagnostic approach to pulmonary embolism with spiral CT, with special emphasis on protocol parameters and scan interpretation. PMID- 9853220 TI - Expiratory CT in cigarette smokers: correlation between areas of decreased lung attenuation, pulmonary function tests and smoking history. AB - The aim of this study was to determine the correlation between cigarette-smoke related bronchial disease and air trapping as assessed by expiratory high resolution CT (HRCT) scans. Thirty healthy subjects (11 non-smokers, 7 ex-smokers for > 2 years, 12 current smokers; age range 35-55 years) with a smoking history between 0 and 28.5 pack-years underwent pulmonary function tests (PFT) and HRCT in inspiration and expiration in supine and prone position. The extent of air trapping was scored in ventral and dorsal aspects of the upper, middle and lower lung portions. In 24 subjects (7 non-smokers, 7 ex-smokers, 10 current smokers) areas of focal air trapping were found, and were present significantly more often in dependent lung portions (p < 0.05) compared with non-dependent portions. No significant differences were found between apical and basal lung zones. Scores of focal air trapping were not significantly different between smokers and ex smokers, but were significantly lower (p < 0.05) in non-smokers and showed a significant (p < 0.0005) correlation with pack-years. The degree of air trapping was also associated with several lung function tests, especially RV, DLCO, FRC, FEV1 and FEV1/VC. Air trapping is seen in smokers with normal PFT and correlates with the severity of the smoking history, independently of current smoking status. PMID- 9853221 TI - Bronchial carcinoid tumor: demonstration by dynamic inversion recovery turbo flash MR imaging. AB - A 38-year-old woman was referred to our hospital following the discovery of a right hilar mass on chest radiograph. Retrospectively, a hilar mass could be seen on a chest X-ray which had been obtained 5 years earlier. A coronal dynamic inversion recovery turbo-FLASH gadolinium-enhanced sequence was performed, demonstrating the right lesion which enhanced during the systemic arterial phase indicating an arterial supply from the bronchial arterial circulation. The surgical and pathological findings were a bronchial carcinoid tumor, with foci of bone formation. PMID- 9853222 TI - Pulmonary arteriovenous fistula: detection with magnetic resonance angiography. AB - Pulmonary arteriovenous fistula are an uncommon disorder, and are most frequently congenital, usually then associated with hereditary hemorrhagic telangectasia (Rendu-Osler-Weber disease). We present, to our knowledge, the first case of a pulmonary arteriovenous fistula detected by gadolinium-enhanced pulmonary magnetic resonance angiography and confirmed by digital subtraction pulmonary angiography in a patient where the CT scan was unremarkable. PMID- 9853223 TI - Laparoscopic ultrasonography for abdominal tumor staging. AB - In recent years, laparoscopic ultrasonography has been introduced as an adjunct to diagnostic laparoscopy for staging of tumors of the upper gastrointestinal tract, liver, biliary tree, and pancreas. It has proved feasible to visualize most anatomic structures in the upper abdomen consistently and in detail with laparoscopic ultrasonography. Recent publications indicate that laparoscopic ultrasonography may be useful for detecting small liver metastases, lymph node metastases, small primary tumors of the pancreas and bile ducts, and for the assessment of the local extension of tumors of the pancreas and stomach. The ongoing improvements in US technology and the results of larger studies will in the near future determine the precise place of this new imaging modality for staging of abdominal tumors. PMID- 9853224 TI - Acute abdomen due to torsion of wandering spleen: CT diagnosis. AB - Two adults and a child with acute abdomen and surgically confirmed torsion of wandering spleen are presented. Computed tomography provided a spectrum of findings including an ovoid or comma-shaped abdominal mass, hypertrophy of the liver's left lobe, a whirled appearance of hyperdense, nonenhancing splenic vessels, and an enlarged spleen, exhibiting minimal or no enhancement. Computed tomography also indicated the point of torsion and the viability of splenic parenchyma. PMID- 9853225 TI - Diameter of the common bile duct in the elderly patient: measurement by ultrasound. AB - We wanted to assess the width of the common bile duct (CBD) in an asymptomatic elderly (> 75 years) population, since no previous report has particularly considered the important age-related increase in CBD diameter for this patient group. CBD width of 92 asymptomatic people over 75 years of age (mean 84.7 years, range 75-96 years) without clinical or laboratory evidence for biliary, hepatocellular or pancreatic disease was measured by real-time high-resolution ultrasound of the upper abdomen. All examinations were performed by one experienced radiologist. Mean values were calculated for three groups: patients with and without cholecystolithiasis and after cholecystectomy. Statistical analysis was carried out by analysis of variance. Considering the measurements of all patients, the mean width (+/- SD) of the CBD was calculated as 6.5 +/- 2.5 mm. The difference between patients without cholecystolithiasis (6.2 +/- 2.3 mm) and patients after cholecystectomy (8.7 +/- 2.9 mm) was statistically significant (P < 0.0001). Cholecystolithiasis (6.0 +/- 1.6 mm) proves not to be an additional factor for CBD widening in comparison with patients without stone disease. The CBD of these very elderly subjects shows a considerable increase in comparison with the recommended borderline values in the ultrasound literature. An upper limit of 10 mm seems reasonable for patients with and without cholecystolithiasis. A significant increase in CBD width after cholecystectomy was found and measurements up to 14 mm may occur. Measurements have to be correlated with clinical and laboratory findings. PMID- 9853226 TI - Transcatheter embolization of symptomatic hepatic venous malformations. AB - The aim of this study was to evaluate our results in treatment and management of symptomatic hepatic venous malformations using transcatheter embolization therapy. From 1991 to 1997 five venous malformations were embolized in one man and four women ranging in age from 31 to 50 years. All patients presented nonspecific abdominal pain and were assessed in the general surgery unit. Percutaneous embolization was recommended. In all cases polyvinyl alcohol was used to embolize the afferent arterioles. Clinical and echographic control follow up was carried out on an outpatient basis in our Vascular and Interventional Radiology Unit. Embolization was possible in all cases. No complications developed during the procedure, except in one case where the subject suffered a spasm of the hepatic artery. Mean follow-up time was 42 months (6-73 months). Four patients remained asymptomatic during the entire follow-up period, whereas one patient required reembolization. In our experience, transcatheter embolization of hepatic venous malformations is a noninvasive technique which requires few admissions and presents few complications. With further use, it could become the treatment of choice in symptomatic hepatic venous malformations as an alternative to surgery. PMID- 9853227 TI - Veno-occlusive disease of the colon--CT findings. AB - Venous occlusion is a rare cause of ischemic bowel disease and is usually brought about by thrombosis that may occur as a complication of systemic disorders like systemic lupus erythematosus. Behcet disease or Churg-Strauss syndrome. This report describes a patient with veno-occlusive disease of the colon caused by lymphocytic phlebitis. Typical CT findings included homogeneous bowel wall thickening and vascular engorgement. PMID- 9853228 TI - Omental fibroma: CT and US findings. AB - The US, CT, and X-ray findings in a patient with omental fibroma of the lesser omentum are described. Ultrasound showed a solid mass with cystic areas in the central region. At CT the lesion showed peripheral enhancement and central hypodensity. On X-ray studies with barium, there was border distortion in the lesser curvature of the stomach. The mass was resected surgically. A pathologic diagnosis of fibroma was confirmed. PMID- 9853229 TI - Primary choriocarcinoma of the stomach and pancreas: CT findings. AB - In this report we present the CT findings of two non-gestational, extragonadal choriocarcinomas, one arising within the stomach and one in the pancreas. These are rare tumours and a pancreatic primary site has not been previously described. PMID- 9853230 TI - Acute obstruction of the renal collecting system: the intrarenal resistive index is a useful yet time-dependent parameter for diagnosis. AB - The aim of this study was to determine whether the intrarenal resistive index (RI) can be used for the diagnosis of acute obstruction in patients with renal colic and to determine whether the index is time-related. Seventy patients referred to the Emergency Department with acute renal colic and without known associated renal disease underwent duplex Doppler ultrasonography to determine the intrarenal RI at the symptomatic and asymptomatic side. The age range of the patients was 18-72 years. An RI greater than 0.68 and/or an interrenal difference in RI greater than 0.06 and/or an increase in RI of more than 11% compared with the normal side proved reliable cut-off values to diagnose acute renal obstruction. In addition, time dependency of the increase in RI was noted. No significant differences were observed within the first 6 h after the onset of symptoms. From 6 to 48 h, however, the mean RI in the affected kidney (0.70 +/- 0.06; mean +/- SD) was significantly different from that in the normal kidney (0.59 +/- 0.04) (P < 0.001). In the same period the mean difference in RI was 0.08-0.13 (P < 0.001). After 48 h the sensitivity of RI dropped substantially. It is concluded that renal duplex Doppler ultrasonography is useful for diagnosing acute renal obstruction between 6 and 48 h after the onset of symptoms. PMID- 9853231 TI - MRI of the uterus, uterine cervix, and vagina: diagnostic performance of dynamic contrast-enhanced fast multiplanar gradient-echo imaging in comparison with fast spin-echo T2-weighted pulse imaging. AB - The aim of this study was to compare the diagnostic performance of contrast enhanced fast multiplanar gradient-echo (GRE) and T2-weighted fast spin-echo (FSE) image sets in the assessment of uterus, cervix, and vagina. Fast (up to 20 contiguous sections in 23 s) multiplanar GRE and FSE images of 45 patients referred for imaging of the female pelvis were evaluated retrospectively with regard to overall image quality and the ability to detect normal anatomic structures, as well as lesion conspicuity. Results were compared with histologic findings (n = 29) or clinical follow-up. Furthermore, a quantitative assessment of contrast-to-noise ratios among normal uterine and cervical structures as well as uterine lesions was performed for both sequences. On GRE images, uterine and cervical differentiation was best seen on the image sets acquired 15 and 60 s following contrast enhancement and results were significantly better compared with delayed images (p < 0.05). Delineation of the junctional zone was significantly (p < 0.05) better on FSE compared with GRE images; no significant difference was seen for the other anatomic structures. Overall image quality of GRE and FSE images was similar. Sensitivity for lesion detection based on both GRE and FSE images was 96% with a sensitivity of 93% for GRE, and 81% for FSE images alone, respectively. Using the extended McNemar chi 2 test, the difference in diagnostic performance between FSE and GRE revealed no significant difference, whereas the combination of both techniques performed better than FSE imaging alone (p < 0.05). The presented data suggest that dynamic contrast-enhanced GRE imaging should be part of an MR examination of the female pelvis. Combined GRE and FSE imaging provide an excellent sensitivity in the assessment of uterine and vaginal pathologies. PMID- 9853232 TI - Metastatic RCC arising in a transplant kidney. AB - We report a case of metastatic renal cell carcinoma arising in a cadaver transplant kidney 6 years after transplantation. Due to molecular analysis of the tumor tissue we could prove that the carcinoma originated from the male donor. After tumor resection and interruption of immunotherapy, the concomitant bone and lymph node metastases resolved with alpha-interferon and interleukin-2-based immunotherapy. PMID- 9853233 TI - Primary leiomyoma of the ovary: a case report. AB - The imaging features of primary ovarian leiomyoma have seldom been described because of the very low prevalence of the tumor. Herein we present a case report on its magnetic resonance imaging features. Both T1- and T2-weighted images revealed a low-signal-intensity mass, which was well circumscribed and sharply demarcated from the uterus. Magnetic resonance imaging was useful in distinguishing it from a malignant ovarian tumor and uterine leiomyoma. PMID- 9853234 TI - Cranial CT revisited: do we really need contrast enhancement? AB - The aim of this study was to define guidelines for intravenous contrast administration in cranial CT, as currently there are no recent guidelines based on a large series of patients. In 1900 consecutive patients (1480 adults and 420 children) pre- and post-contrast scan was analysed in order to assess the contribution of contrast enhancement to the diagnosis. The findings were grouped according to whether abnormalities were seen on the pre- and/or post-contrast scan, or whether no abnormalities were seen at all. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy of a pre contrast scan were used to determine validity. Intravenous contrast enhancement only contributes to the diagnosis if a suspicious abnormality is seen on the unenhanced scan or in the appropriate clinical setting (33.6%). In the remaining patients (65.6%) there is no diagnostic contribution, except for a small number of abnormalities (0.8%). These are often anatomical variants and have no therapeutic impact. The number of contrast-enhanced cranial CT examinations can significantly be reduced by using four general guidelines for contrast administration resulting in considerable cost savings without affecting the quality of service to the patient. These guidelines are defined by the clinical findings/presentation or by the findings on the unenhanced scan. The number of contrast-related complications will be reduced, which may have medicolegal implications. These guidelines can be applied in any radiology department. PMID- 9853235 TI - Magnetic resonance epidurography with gadolinium-DTPA. AB - The aim of this study was to evaluate and describe MRI epidurography as a new imaging tool. Five volunteers and one patient were investigated with MR epidurography after injection of 20 ml Gd-DPTA solution (1:250/1 ml Gd-DPTA/250 ml normal saline). Magnetic resonance epidurography is possible. With fat suppression techniques, the contrast between Gd-DPTA solution in the epidural space and surrounding soft tissue proved adequate. Using the multiplanar capability of MRI with MR epidurography coronal and sagittal projections similar to conventional epidurography, axial slices comparable to CT epidurography can be obtained. Magnetic resonance epidurography is superior to conventional and CT epidurography. Presently, due to high costs as compared with conventional and CT epidurography, MRI is not suitable for the routine monitoring of peridural catheters, but it may have a place in the future with decreasing costs for MRI and for the evaluation of patients with spine pathology, especially in describing epidural processes. PMID- 9853236 TI - Chronic intussusception in childhood. AB - A rare case of chronic intussusception is reported. Radiological investigation including ultrasound, CT and barium enema provided the correct diagnosis. Ultrasound revealed a solid mass near the transverse colon in the right lower abdomen. CT demonstrated the real nature of the solid mass as being the classical 'coiled spring' of intussusception. The barium enema was unsuccessful as an attempt at hydrostatic reduction, but confirmed the diagnosis by a 'crescent shaped' filling defect in the ascending colon. The patient was discharged after an uneventful surgery and recovery. No organic lesion that precipitated the chronic intussusception was discovered at operation. Radiological findings, the typical clinical picture and adequate treatment are discussed. PMID- 9853237 TI - Application of M-mode sonography to functional evaluation in pediatric patients. AB - To assess the feasibility and advantages of functional motion-mode (M-mode) sonography in pediatric patients with various non-cardiac M-mode applications, 50 patients aged 6 days to 14.5 years with megaureter, diaphragmatic palsy and spinal cord malformation were studied. In addition to initial conventional brightness-mode gray-scale ultrasound. M-mode sonography was performed for evaluation of motion and possible impairment. The findings were compared with other imaging results. The impact of the sonographic results on clinical management was evaluated retrospectively. Impaired motion was demonstrated by conventional sonography in all cases. Only M-mode sonography enabled adequate documentation for comparison with follow-up examinations, but allowed only semiquantitative assessment. Thus even gradual improvement or deterioration can be followed in an unbiased way by using a better-documented investigation for comparison, though an objective 'ranking', especially of diaphragmatic movement, could not be achieved. M-mode sonography enables a non-invasive and non-ionizing evaluation and documentation of motion impairment in pediatric patients. It improves documentation of motion and enables a better comparison with previous findings. It is especially useful for follow-up in evolving disease. PMID- 9853238 TI - Differentiation of benign and malignant tumors of the parotid gland: value of pulsed Doppler and color Doppler sonography. AB - To establish criteria for the differentiation of benign and malignant tumors of the parotid gland using color Doppler sonography (CDS) and pulsed Doppler sonography (PDS) we examined 37 patients with parotid tumors by gray-scale ultrasound, CDS, and PDS. Tumor vascularization displayed by CDS was graded subjectively on a 4-point scale (0 = no vascularization, 3 = high vascularization). From the Doppler spectrum, the highest systolic peak flow velocity, the resistive index (RI), and the pulsatility index (PI) were calculated. There were 11 malignant and 26 benign tumors. Tumor vascularization by CDS was grade 0 or 1 in 88.5% of benign lesions, whereas it was grade 2 or 3 in 82% of malignant lesions (P < 0.0001). The highest systolic peak flow velocity was statistically significantly higher in malignant lesions than in benign lesions. Using a threshold systolic peak flow velocity of 25 cm/s, sensitivity was 72% and specificity was 88% for the detection of a malignant tumor. Evaluation of tumor vascularization by CDS and PDS cannot differentiate between benign and malignant parotid tumors with certainty. However, high vascularization and high systolic peak flow velocity in tumor vessels should raise the suspicion of malignancy, even if tumor morphology on gray-scale sonography indicates a benign lesion. PMID- 9853239 TI - The impact of stereotactic large-core needle biopsy in the treatment of patients with nonpalpable breast lesions: a study of diagnostic accuracy in 510 consecutive cases. AB - The objective of this study was to assess the usefulness of stereotactic large core needle biopsy (LCNB) in the management of nonpalpable breast lesions (NBL) and compare it with stereotactic fine-needle aspiration biopsy (SFNA) performed simultaneously in a significant number of cases. From November 1993 through June 1997, 510 consecutive patients with NBL underwent 14-gauge LCNB with 354 women undergoing simultaneous 21-gauge SFNA in the same lesson. Mammographic findings lesion size, number of core biopsy specimens, complications and diagnoses of both techniques were analysed. Surgical biopsy, tumorectomy or mastectomy was indicated for malignancy or poor correlation between SFNA or LCNB results and clinical or radiological findings. Values of diagnostic accuracy of both LCNB and SFNA were determined. The ratio benign surgical biopsies/malignant surgical biopsies (BB/CB) of the series was calculated. A total of 171 patients underwent surgical treatment: in 31 (18.1%) a benign process or atypical ductal hyperplasia was the final diagnosis. The ratio BB/CB was 0.22. Sensitivity and specificity were 93.2 and 100%, respectively, for LCNB, and 77.2 and 92.3%, respectively, for SFNA with cytological analysis. Large-core needle biopsy provides more accurate diagnosis than SFNA in the management of nonpalpable breast lesions and obviates a surgical diagnostic procedure in a significant number of cases. PMID- 9853240 TI - Imaging of tuberculous disease involving breast. AB - The aim of our study was to evaluate radiologic findings of the tuberculosis involving breast. We evaluated the radiologic features of 17 patients (18 lesions) with tuberculous disease involving the breast. The radiologic examinations, including mammography (16 patients), ultrasonography (12 patients), and Gd-DTPA-enhanced dynamic MRI (6 patients), were analyzed. Mammographic findings included mass (12 of 17 lesions), calcification (3 of 17 lesions), asymmetric density with spiculated margin (5 of 17 lesions), and axillary lymph node enlargement (8 of 17 lesions). On ultrasonography, a smooth bordered mass (7 of 13 lesions) with thin boundary (7 of 13 lesions) and heterogeneous, intermediate internal echoes (9 of 13 lesions) were most commonly demonstrated. On Gd-DTPA-enhanced dynamic MRI, 3 lesions showed significant enhancement at the first minute after injection (3 of 7 lesions). The maximum enhancing amount was greater than 500 normalized units, and the enhancing pattern was smooth or irregular ring appearance. Breast involvement with tuberculosis is rare but should be considered in the differential diagnosis of a woman living in an endemic area or when extramammary foci of tuberculosis are present. A multimodality imaging approach with clinical evaluation will help to establish the diagnosis of tuberculosis involving breast. PMID- 9853241 TI - Positron emission tomography as a diagnostic tool in oncology. AB - Early diagnosis in oncology is important for treatment by surgical intervention, which generally has the highest curative potential. For higher stages of disease involvement, initiation of rapid treatment is indicated to provide the patient with the optimal therapy regimen. Although this may not improve the prognosis, it will maintain the quality of life. Anatomic imaging modalities, such as CT, MR imaging, and US, are clinically important high-resolution imaging techniques that are well suited to reveal structural abnormalities. However, the differentiation of lesions as being benign or malignant is still problematic. Metabolic imaging modalities in nuclear medicine (NM), i.e., single photon emission computed tomography (SPECT) and positron emission tomography (PET), can reveal biochemical parameters of the lesions such as glucose, oxygen, or amino acid metabolism, or measure the receptor density status. These parameters may allow a completely new clinical perspective in the management and understanding of diseases such as cancer. Although PET has been around since the early 1960s, it has only recently emerged as a powerful diagnostic tool in oncology. Society has great difficulty accepting this clinical imaging modality because of its high cost and complexity. Current applications of PET in oncology have been in characterizing lesions, differentiating recurrent disease from treatment effects, staging tumors, evaluating the extent of disease, and therapy monitoring. Here, the role of PET in diagnosis, staging, and restaging of cancer is reviewed and compared with the other tumor imaging modalities. We cover articles published in the past 3 years. We utilize the typical radiology format, in which the contribution in each body area is reviewed (topographic orientation), instead of the more organ-based approach used in internal medicine. PMID- 9853242 TI - Tossed on the horn of humerus: a legal case report. AB - Radiological cases constituted 2% of the total number of legal cases in Sweden during 1996. The young man in this report accused a radiologist of having missed a fracture 7 years earlier based on a new radiology examination and report. In reality the latter report was incorrect and the patient had a rare exostosis that explained the clinical signs and symptoms. The alleged fracture was an epiphyseal line to a non-unified ossification centre at the tip of the unique exostosis. PMID- 9853243 TI - Quiz case of the month. The microscopic features defined this pancreatic mass as a macrocystic variant of serous cystadenoma. PMID- 9853244 TI - Pneumatosis intestinalis in AIDS-associated chronic intestinal cryptosporidiosis: a benign course in a severe-looking disease. PMID- 9853245 TI - The role of the radiologist. PMID- 9853246 TI - Treatment of acute renal failure. AB - Acute renal failure is a life threatening illness whose mortality has remained high since the introduction of hemodialysis 25 years ago, despite advances in supportive care. Acute renal failure is an extremely morbid and costly disorder with a significant proportion of patients progressing to end-stage renal disease requiring dialysis. To the nephrologist, acute renal failure remains an extremely frustrating disease, because the pathophysiology is not well understood and the limited therapeutic options force the nephrologist to sit on the sidelines and wait for renal function to return. For example, dialysis remains the only FDA approved treatment for acute renal failure, but dialysis may also cause renal injury that prolongs renal failure. The purpose of this perspective is to understand the results of the recent, largely negative, clinical trials in view of recent advances in the epidemiology of ARF. This review will also discuss diagnostic tools, strategies for improved design of clinical trials, and other therapeutic interventions that will be needed to properly treat acute renal failure in the 21st century. PMID- 9853247 TI - Architectural patterns in branching morphogenesis in the kidney. AB - During kidney development, several discrete steps generate its three-dimensional pattern including specific branch types, regional differential growth of stems, the specific axes of growth and temporal progression of the pattern. The ureteric bud undergoes three different types of branching. In the first, terminal bifid type, a lateral branch arises and immediately bifurcates to form two terminal branches whose tips induce the formation of nephrons. After 15 such divisions (in humans) of this specifically renal type of branching, several nephrons are induced whose connecting tubules fuse and elongate to form the arcades. Finally, the last generations undergo strictly lateral branching to form the cortical system. The stems of these branches elongate in a highly regulated pattern. The molecular basis of these processes is unknown and we briefly review their potential mediators. Differential growth in three different axes of the kidney (cortico-medullary, dorsoventral and rostro-caudal) generate the characteristic shape of the kidney. Rapid advances in molecular genetics highlight the need for development of specific assays for each of these discrete steps, a prerequisite for identification of the involved pathways. The identification of molecules that control branching (the ultimate determinant of the number of nephrons) has acquired new urgency with the recent suggestion that a reduced nephron number predisposes humans to hypertension and to progression of renal failure. PMID- 9853248 TI - Genetic polymorphisms of the renin-angiotensin system and the outcome of focal segmental glomerulosclerosis in children. AB - BACKGROUND: The clinical course of primary focal segmental glomerulosclerosis (FSGS) in children is variable, with some patients having a much more rapidly progressing course than others. The purpose of our study was to compare the frequency of three polymorphisms of the renin-angiotensin system (RAS) in children with FSGS with that in healthy controls of matching ethnic groups, and to determine whether the clinical outcome of FSGS was associated with different RAS genotypes. METHODS: Three RAS genotypes were examined in 47 Jewish and Arab children with biopsy-proven primary FSGS and in a large control group: the ACE insertion/deletion polymorphism in intron 16, the M235T mutation in the angiotensinogen gene, and the A1166C in the angiotensin II type 1 receptor gene (AT1R). RESULTS: Arab patients showed a greater tendency towards progressive renal disease than their Jewish counterparts (12 of 21 vs. 9 of 26, P = 0.05) and were less likely to achieve remission (3 of 21 vs. 11 of 26, P < 0.04), despite similar clinical presentation, medical management and follow-up. The RAS allele prevalence was similar among patients and controls of matching ethnic backgrounds, and no difference in allele frequency was found between Arabs and Jews. Homozygotes for the ACE insertion genotype (II) were significantly less likely to have progressive renal disease than patients with the other genotypes (ID and DD; 0 of 6 vs. 21 of 41; P < 0.022). The other RAS polymorphisms were not associated with variations in the clinical course of childhood FSGS. CONCLUSIONS: Homozygosity for the ACE insertion allele may have a protective effect in children with FSGS and can serve as a positive prognostic indicator at diagnosis. The D allele may exert a detrimental dominant effect on outcome. Neither the ACE gene polymorphism nor the other RAS polymorphisms studied are associated with disease prevalence. The AT1R and angiotensinogen gene polymorphisms are not associated with progression of renal disease in FSGS. Ethnic differences in the clinical course of the disease are not linked to these polymorphisms. PMID- 9853250 TI - Role of distinct type IV collagen networks in glomerular development and function. AB - BACKGROUND: In X-linked Alport syndrome, mutations in the COL4A5 gene encoding the alpha 5 chain of type IV collagen result in progressive renal failure. This nephropathy appears to relate to the arrest of a switch from an alpha 1/alpha 2 to an alpha 3/alpha 4/alpha 5 network of type IV collagen in the developing glomerular basement membrane (GBM; Kalluri et al, J Clin Invest 99:2470, 1997). METHODS: We examined the role of this switch in glomerular development and function using a canine model of X-linked nephritis with a COL4A5 mutation. The electron microscopic appearance and the expression of the alpha 1-alpha 6 chains of type IV collagen in the GBM was correlated with glomerular function. RESULTS: In normal neonatal glomeruli, once capillary loops were present, there was staining of GBM for the alpha 1-alpha 5 chains. Prior to this stage, only alpha 1 and alpha 2 chains were present, with rare glomeruli positive for the alpha 5 chain. As glomeruli matured, the alpha 1 and alpha 2 chains tended to disappear from the GBM, with the alpha 3-alpha 5 chains remaining. In affected male dogs, only the alpha 1 and alpha 2 chains were detected at any stage. GBM ultrastructure in these dogs remained normal until one month and proteinuria did not appear until two months. CONCLUSION: Our results show that normal glomerular development involves a switch in type IV collagen networks. In affected male dogs, a failure of this switch results in an absence of the alpha 3/alpha 4/alpha 5 network and a persistence of the alpha 1/alpha 2 network in GBM. GBM ultrastructure and glomerular function remain normal for one month, indicating that GBM deterioration in Alport syndrome begins as a postnatal process. Hence, only the alpha 1/alpha 2 network is essential for normal glomerular development, whereas the alpha 3/alpha 4/alpha 5 network is essential for long-term maintenance of glomerular structure and function. PMID- 9853249 TI - Functional characterization of renal chloride channel, CLCN5, mutations associated with Dent'sJapan disease. AB - BACKGROUND: The annual urinary screening of Japanese children above three years of age has identified a progressive renal tubular disorder characterized by low molecular weight proteinuria, hypercalciuria and nephrocalcinosis, and this represents a variant of Dent's disease. Hitherto, 12 mutations of the X-linked renal specific chloride channel, CLCN5, have been reported in the Dent'sJapan variant. To further identify such CLCN5 mutations and to define the structure function relationships of this channel, we have investigated five unrelated, non consanguinous Japanese families with this disorder. METHODS: Leukocyte DNA from probands was used with CLCN5 primers for PCR amplification of the coding region, and the DNA sequences of the products determined. Functional studies were performed by expressing the mutants in Xenopus oocytes. RESULTS: Five CLCN5 mutations consisting of two nonsense (R648X and R704X), two missense (S270R and L278F) and one acceptor splice site mutation (ag-->cg) in intron 4 were identified. The missense and splice site mutations represent novel abnormalities. Heterologous expression in Xenopus oocytes of wild-type and the missense mutants demonstrated that the mutations, which were translated, either abolished or markedly reduced chloride conductance. CONCLUSIONS: These results expand the spectrum of CLCN5 mutations associated with this renal disorder and provide insight into possible structure-function relationships. For example, both the missense mutations are located within a short putative loop between two transmembrane domains, and our results suggest that this region may have an important functional role in the regulation of channel activity. PMID- 9853251 TI - Platelet-activating factor acetylhydrolase gene mutation in Japanese nephrotic children. AB - BACKGROUND: Platelet-activating factor (PAF) may be involved in the pathogenesis of steroid-responsive nephrotic syndrome (SRNS). PAF is degraded to inactive products by PAF acetylhydrolase. We have investigated whether PAF acetylhydrolase gene mutation is involved in SRNS in Japanese children. METHODS: We identified a point mutation in the PAF acetylhydrolase gene (G994T) using the polymerase chain reaction in 101 Japanese children with SRNS and 100 healthy Japanese. RESULTS: There was no difference in the genotype and allele frequencies between patients with SRNS and normal controls. The mean number of relapses during the first year after onset was significantly higher in the 26 patients who were heterozygous for the mutant allele (GT) than in 75 wild-type homozygotes (GG) (2.61 +/- 1.98 vs. 1.33 +/- 1.35; P = 0.0019). CONCLUSIONS: We conclude that analysis of the PAF acetylhydrolase gene mutation at position 994 in Japanese children with SRNS allows the identification of patients who are more likely to have a disease relapse. PMID- 9853252 TI - Sequential effects of high glucose on mesangial cell transforming growth factor beta 1 and fibronectin synthesis. AB - BACKGROUND: Transforming growth factor (TGF)-beta is recognized as the final common mediator of the principal lesions of diabetic nephropathy such as renal hypertrophy and mesangial expansion. To gain better understanding of the temporal relationships between high glucose (HG) and mesangial cell (MC) TGF-beta 1 synthesis and between TGF-beta 1 and extracellular matrix (ECM) synthesis, the present study examined early and sequential effects of HG on TGF-beta 1 and fibronectin (FN) mRNA expression and protein synthesis. METHODS: Confluent primary rat MC was stimulated with 5.6 (control) or 30 (high) mM glucose after synchronizing the growth by incubation with serum-free media for 48 hours. RESULTS: Mesangial cell TGF-beta 1 mRNA expression increased significantly in six hours and continued to increase until 48 hours in response to HG. The level of TGF-beta 1 mRNA was 1.5-fold higher than that of control glucose at six hours and 1.8-fold at 48 hours. TGF-beta activity in heat-activated conditioned media under HG increased 1.5- and 1.6-fold at 24 and 48 hours, respectively, compared to control glucose. FN mRNA increased significantly at 24 and 48 hours and 1.4-fold that of control glucose at both time points. FN protein also increased 1.5-fold that of control glucose at 48 hours. Anti-TGF-beta antibody completely abolished HG-induced FN synthesis. CONCLUSIONS: The present finding demonstrate that HG stimulates TGF-beta 1 very early and prior to FN production and that HG-induced FN production is mediated by TGF-beta. This finding is consistent with the view that TGF-beta mediates increased ECM accumulation by MC under high glucose conditions. PMID- 9853253 TI - Growth of long bones in renal failure: roles of hyperparathyroidism, growth hormone and calcitriol. AB - BACKGROUND: The treatment of secondary hyperparathyroidism (2 degrees HPT) associated with chronic renal failure adversely affects skeletal growth. METHODS: We assessed epiphyseal growth plate morphology by quantitative histology and measured mRNA levels for selected markers of chondrocyte proliferation and differentiation by in situ hybridization in the growth plate cartilage of subtotally nephrectomized rats with either mild or advanced 2 degrees HPT. RESULTS: The width of the growth plate cartilage in the proximal tibia and mRNA levels for PTH/PTHrP receptor were unchanged in rats with mild 2 degrees HPT, however, they were markedly less in nephrectomized rats with advanced 2 degrees HPT than in intact controls. Treatment with growth hormone 10 IU/kg/day increased growth plate thickness both in mild and in advanced 2 degrees HPT and raised mRNA levels for type II and type X collagen in rats with advanced 2 degrees HPT. The administration of calcitriol 50 ng/kg/day attenuated these responses in animals with advanced 2 degrees HPT. Overall, PTH/PTHrP receptor mRNA levels did not correspond to the serum levels of PTH indicating that PTH/PTHrP receptor expression is down-regulated in renal failure by a PTH-independent mechanism. CONCLUSION: Calcitriol counteracts the trophic actions of growth hormone on epiphyseal growth plate cartilage and modifies chondrocyte differentiation in vivo, and these mechanisms may contribute to disturbances in longitudinal bone growth in renal failure. PMID- 9853254 TI - Differential induction of functional B1-bradykinin receptors along the rat nephron in endotoxin induced inflammation. AB - BACKGROUND: Under physiological conditions, the effects of kinins in the kidney are mainly mediated by the bradykinin B2-receptor, whereas the kinin B1-receptor is strongly induced under inflammatory conditions in a variety of tissues. Knowledge of the distribution of the B1-receptor along the nephron is of importance since the B1-receptor might replace B2-receptors under these conditions. METHODS: Using a RT-PCR/Southern blot approach allowing relative quantification of mRNA levels, ten different microdissected rat nephron segments were analyzed for the presence of the B1- and B2-receptor before and after endotoxin treatment to induce experimental inflammation. The functionality of the expressed receptors was assessed by kinin-induced intracellular calcium ([Ca2+]i) mobilization in microdissected nephron segments. RESULTS: While under physiological conditions no B1-receptor mRNA could be detected, after 18 hours of treatment with bacterial lipopolysaccharide (LPS) the expression of B1-receptor mRNA was strongly induced in the efferent arteriole, the medullary and inner medullary thin limb, and in the distal tubule. Moderate expression was found in the glomerulus, proximal convoluted and straight tubules, and in the medullary thick ascending limb. Small but detectable expression was observed in the cortical collecting duct. The induction of B1-receptor mRNA expression resulted in functional receptor expression, since increases in [Ca2+]i were observed upon B1-agonist stimulation. LPS treatment also increased the expression of B2 receptor mRNA in all nephron segments except in the glomerulus, the inner medullary thin limb and the outer medullary collecting duct. However, no related changes in B2-agonist induced rises in [Ca2+]i were found. CONCLUSIONS: These studies show a functional induction of the B1-kinin receptor along the rat nephron, which should be taken in account to address the effects of kinins under inflammatory conditions in the kidney. PMID- 9853255 TI - Localization of the prostacyclin receptor in human kidney. AB - BACKGROUND: Prostacyclin is an important mediator of renal hemodynamics. Furthermore, recent studies argue for a role of this arachidonic acid metabolite in the regulation of salt and water handling in the distal nephron. To gain insight into the network of prostacyclin signal transduction, we analyzed the intrarenal distribution of the prostacyclin receptor (IP receptor) in adult human kidney. METHODS: Specific polyclonal antibodies against a synthetic peptide of the human IP receptor were generated. By means of immunohistology the localization of IP receptor protein was studied. The mRNA expression for IP receptor was analyzed by in situ hybridization using specific cRNA probes. RESULTS: In human kidney sections both IP receptor-immunoreactive protein and mRNA were expressed in smooth muscle cells and endothelial cells. Expression of the IP receptor was observed in glomerular cells, namely mesangial cells, endothelial cells, and podocytes. Both mRNA and protein expression for IP receptor was observable in Tamm-Horsfall-negative distal tubules and collecting ducts. CONCLUSIONS: The vascular expression of the IP receptor is consistent with the known vasodilatory effect of prostacyclin in vascular beds. Glomerular expression argues for a role of this autacoid in the regulation of glomerular hemodynamics. The tubular distribution might point towards the involvement of prostacyclin in renal salt and water handling. PMID- 9853256 TI - AVPR2 variants and V2 vasopressin receptor function in nephrogenic diabetes insipidus. AB - BACKGROUND: The AVPR2 gene encodes the type 2 vasopressin receptor, a member of the vasopressin/oxytocin receptor subfamily of G protein-coupled receptors. Disruption of AVPR2 causes X-linked congenital nephrogenic diabetes insipidus (NDI), yet the functional significance of most gene sequence variations found in association with NDI has not been proven. The large number of naturally occurring AVPR2 mutations constitutes a model system for studying the structure-function relationship of G protein-coupled receptors. This analysis can be aided by examining amino acid sequence variation and conservation among evolutionarily disparate members of the subfamily. METHODS: Twenty-five new NDI patients were evaluated by DNA sequencing for mutations in AVPR2. Receptors encoded by eighteen NDI alleles were tested for physiologic signaling activity in response to varying concentrations of arginine vasopressin (AVP) in a sensitive cell culture assay. Seventeen amino acid sequences from the vasopressin/oxytocin receptor subfamily were aligned and conserved residues were identified and correlated with the locations of NDI associated variations. RESULTS: Twenty-four variant alleles were found among the 25 new patients. Thirteen had no prior family history of expressed NDI. All 18 of the NDI-associated AVPR2 alleles tested for function demonstrated diminished response to stimulation with AVP. Twelve failed to respond at all, whereas six signaled only at high AVP concentrations. Evolutionarily conserved residues clustered in the transmembrane domains and in the first and second extracellular loops, and NDI-associated missense mutations appeared mostly in the conserved domains. CONCLUSIONS: Sporadic cases are frequent and they usually represent the X-linked rather than the autosomal form of NDI. Genetic and functional testing can confirm this in individual cases. Mutations in this study affecting ligand binding domains tend to retain partial signaling in vitro, whereas those that introduce a charged residue in a transmembrane domain are inactive. The minimal partial signaling observed in cultured cells is unlikely to correlate with clinically significant urine concentrating ability. Other AVPR2 mutations with milder effects on receptor function probably exist, but may not be expressed clinically as typical NDI. PMID- 9853257 TI - Angiotensin II-mediated expression of p27Kip1 and induction of cellular hypertrophy in renal tubular cells depend on the generation of oxygen radicals. AB - BACKGROUND: Angiotensin II (Ang II) induces hypertrophy of cultured proximal tubular cells. We have previously demonstrated that this Ang II-mediated hypertrophy occurs in the G1-phase of the cell cycle and depends on the induction of p27Kip1, an inhibitor of G1-phase cyclin/cyclin-dependent kinase complexes. The present study was undertaken to investigate whether Ang II may stimulate superoxide anions (O2.) formation in cultured LLC-PK1 and cultured mouse proximal tubule (MCT) cells, and to gain further insight into a potential relationship between O2. and cell cycle regulation. METHODS: Reactive oxygen species were measured with the lucigenin method in intact cells. The effects of various inhibitors were tested on Ang II-induced O2. production. Cells were transiently transfected with phosphorothioate-modified rat p22phox antisense oligonucleotides to investigate the potential role of NAD(P)H oxidase. Expression of p22phox mRNA after Ang II-treatment was detected with Northern blots. Incorporation of [3H]leucine into de novo synthesized proteins was used as a parameter of cell hypertrophy. Expression of p27Kip1 was evaluated in cell lysates by Western blotting. RESULTS: Ang II stimulated the accumulation of O2. in tubular cells; however, an addition of two different antioxidants completely abolished measurable O2. This effect was transduced by angiotensin receptor type-1 (AT1) and was inhibited by a flavoprotein inhibitor (DIP) or p22phox antisense oligonucleotides, indicating the involvement of membrane NAD(P)H oxidase. Ang II stimulated de novo protein synthesis was attenuated by DIP, antioxidants, and p22phox antisense oligonucleotides. The Ang II-induced expression of p27Kip1 protein and cellular hypertrophy were reduced by similar treatments. Generation of O2. by xanthine supplementation also stimulated p27Kip1 expression and induced hypertrophy in LLC-PK1 cells. CONCLUSIONS: This study provides the first evidence, to our knowledge, that Ang II induces O2. in cultured tubular cells. Ang II-mediated activation of membrane bound NAD(P)H oxidase, probably by an increase in p22phox transcripts, is likely responsible for this induction. Generation of O2. subsequently induces p27Kip1 expression and stimulates hypertrophy, suggesting a novel mechanism of how Ang II can modulate cell cycle regulation. PMID- 9853258 TI - The plasma membrane-actin linking protein, ezrin, is a glomerular epithelial cell marker in glomerulogenesis, in the adult kidney and in glomerular injury. AB - BACKGROUND: Ezrin belongs to a family of plasma membrane-cytoskeleton linking, actin binding proteins (Ezrin-radixin-Moesin family) involved in signal transduction, growth control, cell-cell adhesion, and microvilli formation. METHODS: The expression of ezrin was examined in glomerular cells in culture, during kidney development, in the mature kidney, and in five different experimental kidney disease models in the rat. RESULTS: Ezrin was specifically expressed in glomerular epithelial cells in developing glomeruli in mature glomeruli and in glomerular epithelial cells in culture. Distinct from its other family members, moesin and radixin, which are predominantly expressed in glomerular endothelial and mesangial areas, ezrin protein (by immunohistochemistry) was specifically and exclusively modulated during podocyte injury and regeneration. Ezrin immunohistochemistry was able to visualize cell body attenuation, pseudocysts, and in particular vacuolation of injured podocytes, a feature that usually has to be identified at the ultrastructural level, and was strikingly increased in binucleated podocytes or podocytes that were partially or completely detached from the underlying GBM (frequently also binucleated). Infiltrating macrophages also express ezrin, but can easily be differentiated from podocytes by their round shape and higher level of expression. CONCLUSIONS: Ezrin likely has a role in the cytoskeletal organization, such as reassembling of acting filaments accompanying podocyte injury and regeneration. Since suitable light microscopic markers for the identification of glomerular epithelial cells are rare, ezrin may also be a useful marker for podocytes in normal and injured glomeruli. PMID- 9853259 TI - Chemokine receptor (CCR5) expression in human kidneys and in the HIV infected macaque. AB - BACKGROUND: The chemokine receptor, CCR5, has been identified as an essential co receptor with CD4, which permits entry of human immunodeficiency virus (HIV) into mammalian cells. This receptor may also mediate leukocyte and parenchymal responses to injury by virtue of its binding to locally released chemokines such as RANTES, MIP-1 alpha and MIP-1 beta during inflammation. The localization of CCR5 in human or primate kidney is unknown. In this study we sought to identify sites of CCR5 synthesis through localization of mRNA coding for this peptide. METHODS: CCR5 cDNA cloned into an expression vector was transcribed into a 1.1 Kb antisense riboprobe that was utilized for in situ hybridization (ISH) and Northern blotting studies. RESULTS: Northern analysis demonstrated positive hybridization for CCR5 mRNA in total RNA isolated from allograft nephrectomy tissue with features of severe transplant rejection as well as in kidney tissue with focal interstitial nephritis. No comparable hybridization signal was achieved with human kidney tissue uninvolved by disease. CCR5 mRNA was not identified in intrinsic renal cell types by ISH in normal human (N = 6), normal macaque kidney (N = 5), in kidneys from macaques with established infection by HIV-2 (N = 9), kidneys from macaques infected with HIV-1 (N = 4), nor in kidneys from SIV-infected macaques (N = 5). CCR5 was identified by ISH in human kidneys with features of interstitial nephritis (N = 3) and in rejected human allograft kidneys (N = 14). The expression of CCR5 was restricted to infiltrating mononuclear leukocytes at sites of chronic tubulointerstitial injury and at sites of vascular and interestitial rejection, respectively. CONCLUSIONS: Understanding the localization of CCR5 as well as other chemokine receptors may help us understand how specificity in leukocyte trafficking is achieved in renal inflammatory processes such as allograft rejection and interstitial nephritis. They provide additional evidence that chemokines may be critical mediators of leukocyte trafficking in renal allograft rejection. These findings may account in part for the difficulty in demonstrating HIV infection of renal cells in human HIV infection, since these cells appear to lack constitutive expression of an essential co-receptor needed for viral entry. PMID- 9853260 TI - Apoptosis and necrosis: mechanisms of cell death induced by cyclosporine A in a renal proximal tubular cell line. AB - BACKGROUND: The mechanisms of cyclosporine (CsA)-induced nephrotoxicity are not fully understood. While hemodynamic changes may be involved in vivo, there is also some evidence for tubular involvement. We previously showed direct toxicity of CsA in the LLC-PK1 renal tubular cell line. In the current study we examined mechanisms (apoptosis or necrosis) of cell death induced by CsA in the LLC-PK1 renal proximal tubular cell line. The possible role of the Fas (APO-1/CD95) antigen-Fas ligand system in the mediation of CsA-induced cell death was also investigated. METHODS: Cells were treated with CsA (0.42 nM to 83 microM) for 24 hours and alterations in DNA and protein synthesis and membrane integrity were examined. Flow cytometry was used to investigate: (i) alterations in the DNA content and cell cycle; (ii) the forward (FSC) and side (SSC) light scattering properties (indicators of cell size and granularity, respectively); (iii) the externalization of phosphatidylserine (PS) as a marker of early apoptosis using FITC-annexin V binding; and (iv) expression of the apoptotic Fas protein. DNA fragmentation in apoptotic cells was also determined by the TUNEL assay. RESULTS: CsA (all doses) caused a block in the G0/G1 phase of the cell cycle as indicated by a decrease in DNA synthesis and supported by an increase in the % of cells in the G0/G1 phase with concurrent decreases of those in the S and G2/M phases. The effect on protein synthesis appeared to be much less. Lower doses of CsA (4.2 nM) caused the appearance of a "sub-G0/G1" peak, indicative of reduced DNA content, on the DNA histogram that was paralleled by a reduction in cell size and an increased cell granularity and an increase in FITC-annexin V binding. DNA fragmentation was evident in these cells as assessed using the TUNEL assay. Higher doses of CsA increased cell size and decreased cell granularity and reduced membrane integrity. Expression of Fas, the cell surface molecule that stimulates apoptosis, was increased following low dose CsA exposure. CONCLUSIONS: These results indicate that CsA is directly toxic to LLC-PK1 cells with reduced DNA synthesis and cell cycle blockade. The mode of cell death, namely apoptosis or necrosis, is dose dependent. Fas may be an important mediator of CsA induced apoptosis in renal proximal tubular cells. PMID- 9853261 TI - Expression of CD27 and ischemia/reperfusion-induced expression of its ligand Siva in rat kidneys. AB - BACKGROUND: Studies identifying genes that are differentially expressed following induction of acute ischemic injury have been useful in delineating the pathophysiology of acute renal failure. METHODS: A differential cDNA library screening technique was used to identify genes that are differentially expressed in rat kidney following induction of acute ischemic renal injury. RESULTS: Levels of mRNA with a high homology to that coding for Siva, a human proapoptotic protein, were increased approximately 4.5-fold in kidneys obtained from rats within 12 hours following ischemia, compared to kidneys from sham-operated rats. A partial cDNA sequence for the rat protein (rat Siva) was determined that overlaps 92% of the human open reading frame. The cDNA sequence predicts a protein 177 amino acids in length with 76% homology to human Siva. Levels of rat Siva in kidneys were elevated at one, five and seven days post-ischemia were not different from those in kidneys from sham-operated controls. In situ hybridization demonstrated that rat Siva mRNA was expressed in cells lining damaged sections in the S3 segment of the proximal tubule at 12 hours and one day post-ischemia. At five and seven days, Siva mRNA was located in epithelial cells of regenerating tubules including in papillary proliferations. TdT-mediated dUTP biotin nick end-labeling (TUNEL)-positive cells colocalized with cells containing Siva mRNA. CD27, the receptor for Siva was localized by immunohistochemistry to sloughed cells in the lumens of damaged S3 segments at 12 hours post-ischemia and to cells within papillary proliferations at five days post-injury. CONCLUSIONS: Siva that is produced within the kidney could be a mediator of apoptosis post ischemia via an interaction with CD27. PMID- 9853262 TI - Soluble latent membrane-type 1 matrix metalloprotease secreted by human mesangial cells is activated by urokinase. AB - BACKGROUND: Matrix metalloprotease 2 (MMP2) is secreted in a latent inactive form (pro-MMP2) that is activated on the cell surface by a membrane-type 1 MMP (MT1 MMP) in the presence of the tissue inhibitor of MMP (TIMP2). In spite of evidence for the synthesis of MT1-MMP shown by immunoblotting, immunocytochemistry and RT PCR, and of TIMP2, MMP2 was found exclusively in a latent form in human mesangial cells (HMC) serum-free culture medium. METHODS AND RESULTS: On purified membranes of HMC, MT1-MMP was found in a 63 kD latent form and as a faint band of 55 kD. The 55 kD band was also present in the ultracentrifuged conditioned medium and likely represented MT1-MMP cleaved from its transmembrane domain, since Northern blot analysis showed only one transcription product. The addition of urokinase plasminogen activator (uPA, 100 nM) to HMC membranes induced the activation of pro-MMP2 via the activation of latent membrane-associated MT1-MMP as reflected by the cleavage of the 63 and 55 kD forms. In addition, when the conditioned medium was successively incubated with uPA and alpha 2-macroglobulin and analyzed by immunoblotting, MT1-MMP decreased, indicating that the soluble MT1-MMP was in a latent form and was activated by uPA. CONCLUSION: Our results provide the first evidence, to our knowledge, of the existence of a soluble latent form of MT1-MMP secreted by primary human cells in culture, confirming that MT1-MMP is an ectoenzyme, and show that uPA can regulate MT1-MMP activity in a soluble phase. PMID- 9853263 TI - Effects of oral antihyperglycemic agents on extracellular matrix synthesis by mesangial cells. AB - BACKGROUND: Increased expression of the glucose transporter GLUT1 in mesangial cells (MCs) markedly stimulates glucose transport and the formation of extracellular matrix (ECM), even when ambient glucose concentrations are low. Certain antihyperglycemic agents cause GLUT1 overexpression and increase glucose transport in various tissues. However, their effects on the kidney are unknown. Because diabetic glomerulosclerosis is characterized by the accumulation of mesangial matrix, was studied the effects of antihyperglycemic agents on matrix metabolism in MCs cultured either in 8 or 20 mM glucose. METHODS: Membrane associated GLUT1 was measured by immunoblotting. The initial rate of glucose transport was determined according to the 2-deoxy-D[14C(U)]glucose uptake. Collagen metabolism was studied by metabolic radiolabeling with [14C]-proline. Fibronectin in the medium was measured by ELISA. GLUT1 mRNA was estimated by Northern analysis. RESULTS: The sulfonylurea tolazamide increased GLUT1 protein expression by 107 and 69% in 8 and 20 mM glucose-grown cells, respectively. However, GLUT1 mRNA levels remained unchanged. Transporter-dependent deoxyglucose uptake was increased by tolazamide up to 184% in a dose-dependent fashion and was evident at both glucose concentrations after three or five days of exposure to the drug. Tolazamide significantly stimulated transforming growth factor-beta 1 (TGF-beta 1) secretion and the total synthesis of collagen and collagen and fibronectin accumulation in the medium of MCs maintained in high or low glucose concentrations. The biguanide metformin did not alter GLUT1 expression, glucose transport, fibronectin formation, or collagen metabolism, except at high concentrations. CONCLUSION: Tolazamide markedly enhances ECM synthesis and accumulation in MCs probably by stimulating GLUT1 expression, glucose transport and TGF-beta 1 secretion, irrespective of the ambient glucose concentration. This effect was dose-dependent and minimally inducible by metformin. PMID- 9853264 TI - Strain differences rather than hyperglycemia determine the severity of glomerulosclerosis in mice. AB - BACKGROUND: We reported that ROP, but not C57, mice were prone to glomerulosclerosis (GS) after nephron reduction (J Clin Invest 97:1242, 1996). METHODS: In this study, we induced diabetes in ROP and C57 mice to determine if the glomerulosclerotic response was stimulus specific. We used the oligosyndactyly mutation (Os), to produce a congenital 50% reduction in nephron number. Stable hyperglycemia was induced by streptozotocin and mice were maintained for 12 weeks without insulin treatment. RESULTS: Glomerular hypertrophy occurred in diabetic ROP +/+ and C57 +/+ mice, but glomeruli of diabetic ROP +/+ mice had 1.92-fold higher laminin B1 and 1.5-fold higher tenascin mRNA levels than diabetic C57 +/+ mice. Diabetic ROP Os/+ mice had severe glomerulosclerosis with arteriolar and tubulointerstitial lesions while there was only moderate mesangial sclerosis in diabetic C57 Os/+ mice. Glomerular size was increased in all non-diabetic Os/+ mice. It was further increased in diabetic ROP Os/+ mice, but not in diabetic C57 Os/+ mice. Glomerular mRNA levels were higher in diabetic ROP OS/+ than in diabetic C57 OS/+ mice [alpha 1 (i.v.) collagen 3.2-fold, laminin B1 2.1-fold, and tenascin 1.6-fold]. CONCLUSION: Overall, our data further support the hypothesis that the susceptibility to glomerulosclerosis is inherited, and suggest that hyperglycemia serves principally as a triggering event in the development of diabetic nephropathy. Since the acceleration of diabetic nephropathy by nephron reduction was also largely strain dependent, it appears that the propensity to glomerulosclerosis is a general renal response and is not stimulus specific. PMID- 9853265 TI - Spontaneous apoptosis of podocytes in explanted glomeruli. Technical note. AB - Despite the extensive use of isolated glomeruli for kidney research, little is understood about whether and how isolation and explantation affect the structure and function of the glomerulus. In this report, we investigate the incidence of apoptosis in explanted, normal rat glomeruli. By using ladder detection assay, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and histological analysis, the present study provides evidence that significant number of resident cells, especially podocytes, spontaneously undergo apoptosis immediately after explantation of glomeruli. PMID- 9853266 TI - Renovascular adaptive changes in chronic hypoxic polycythemia. AB - BACKGROUND: Chronic hypoxia in rats produces polycythemia, and the plasma fraction falls, reducing renal plasma flow (RPF) relative to renal blood flow (RBF). Polycythemia also causes increased blood viscosity, which tends to reduce RBF and renal oxygen delivery. We studied how renal regulation of electrolyte balance and renal tissue oxygenation (which is crucial for erythropoietin regulation) are maintained in rats during hypoxic exposure. METHODS: Rats of two strains with differing polycythemic responses, with surgically implanted catheters in the urinary bladder, femoral artery, and left renal and right external jugular veins, were exposed to a simulated high altitude (0.5 atm) for 0, 1, 3, 14, and 30 days, after which RPF (para-aminohippurate clearance), glomerular filtration rate (GFR, polyfructosan clearance), hematocrit and blood gases were measured, and RBF, renal vascular resistance and hindrance (resistance/viscosity), renal oxygen delivery, and renal oxygen consumption were calculated. RESULTS: During chronic hypoxia RBF increased, but RPF decreased because of the polycythemia. GFR remained normal because the filtration fraction (FF) increased. Renal vascular resistance decreased, and renal vascular hindrance decreased more markedly. Renal oxygen delivery and consumption both increased. CONCLUSIONS: During chronic hypoxia GFR homeostasis apparently took precedence over RBF autoregulation. The large decrease in renal vascular hindrance suggested that renal vascular remodeling contributes to GFR regulation. The reduced hindrance also prevented a vicious cycle of increasing polycythemia and blood viscosity, decreasing RBF, and increasing renal hypoxia and erythropoietin release. PMID- 9853267 TI - Regulation of macula densa Na:H exchange by angiotensin II. AB - BACKGROUND: Angiotensin II (Ang II) is a positive modulator of tubuloglomerular feedback (TGF). At the present time, the site(s) at which Ang II interacts with the signal transmission process remains unknown. In certain renal epithelia, Ang II is known to stimulate apical Na:H exchange. Since macula densa cells possess an apical Na:H exchanger and Ang II subtype I receptors (AT1-receptors), we tested the possibility that Ang II might stimulate exchanger activity in these cells. METHODS: Using the isolated perfused thick ascending limb with attached glomerulus preparation dissected from rabbit kidney, macula densa intracellular pH (pHi) was measured with fluorescence microscopy using BCECF. RESULTS: Control pHi, during perfusion with 25 mM NaCl and 150 mM NaCl in the bath, averaged 7.22 +/- 0.02 (N = 24). Increasing luminal [NaCl] to 150 mM elevated pHi by 0.54 +/- 0.04 (N = 7, P < 0.01). Ang II (10(-9) M), added to the bath in the same paired experiments, significantly elevated baseline pHi by 0.17 +/- 0.04, increased the magnitude of change in pHi (delta = 0.71 +/- 0.05) and initial rate of alkalinization (by 69%) to increased luminal [NaCl]. Ang II produced similar effects when added exclusively to the luminal perfusate. In addition, low-dose Ang II (10(-9) M) stimulated while high-dose Ang II (10(-6) M) inhibited Na dependent pH-recovery from an acid load. AT1 blockade prevented the stimulatory but not the inhibitory effects of Ang II. CONCLUSION: Through the AT1, Ang II may influence macula densa Na transport and regulate cell alkalinization via the apical Na:H exchanger. Thus, Ang II may modulate the TGF signal transmission process, at least in part, through a direct effect on macula densa cell function. PMID- 9853268 TI - A theoretical four-compartment model to evaluate separate kidney technetium-99m MAG3 kinetics in humans. AB - BACKGROUND: Pharmacokinetic modeling based on compartmentalization has provided a valuable tool to assess the clearance patterns of various glomerular and tubular agents. However, no models have been proposed thus far that combine vascular data and imaging data in order to gain a deeper knowledge on renal pathophysiology, and to obtain more diagnostic information of clinical relevance. To this aim, we propose a four-pool model for the evaluation of separate renal function. METHODS: In a group of ten normal volunteers and twenty patients with various renal diseases, we tested the four-pool model based on the identification of the two kidneys as two distinct pools. This approach made it possible to integrate the separate kidney contributions deriving from in vivo imaging data, and allows the researcher to quantitate many parameters specific to each kidney. RESULTS: The parameters TERR, TERL, MRTR, MRTL, vR, vL, k3R-1, K3L-1 permit the normal from abnormal states of renal function to be differentiated, as well as monolateral from bilateral renal disease to be separated within the abnormal function group. CONCLUSIONS: The proposed approach combines the advantages of plasma clearance methods with those derived by gamma-camera imaging, and makes it possible to quantitate the differential renal function. This feature may be clinically relevant in renal transplant donors, where full knowledge of renal pathophysiology could guide the procedure. PMID- 9853269 TI - Nephroprotection by long-term ACE inhibition with ramipril in spontaneously hypertensive stroke prone rats. AB - BACKGROUND: The effect of life-long treatment with the ACE inhibitor ramipril on hypertension-induced histological changes in the kidney was tested in stroke prone spontaneously hypertensive rats (SHR-SP). METHODS: One-month-old pre hypertensive SHR-SP were randomized into three groups of 45 animals each, and exposed via drinking water for their lifetime to a dose of: 1 mg.kg-1.d-1 ramipril (antihypertensive dose, HRA); 10 micrograms.kg-1.d-1 slight dose of ramipril (non-antihypertensive dose, LRA); or placebo. Histological and biochemical assessments were conducted after 15 months in ten rats each, when about 80% of the placebo group had died. RESULTS: Kidneys from placebo treated SHR-SP showed pronounced arterial wall hypertrophy and sclerosis, arterial fibrinoid necrosis, glomerulopathy and tubular interstitial injury that were, in concert with normalized blood pressure, completely prevented by HRA treatment. LRA treatment did not affect any blood pressure increase, and also attenuated the development of arterial wall hypertrophy, sclerosis and arterial fibrinoid necrosis, though to a minor extent only, but did not change glomerular and tubulointerstitial degeneration. These effects of ramipril were associated with a dose-dependent inhibition of plasma and renal tissue ACE activities as well as lower serum concentrations of creatinine, but there were no changes in serum potassium. CONCLUSIONS: Life-long HRA-induced ACE inhibition protects against hypertension-induced renal damages in SHR-SP. This is associated with a doubling of the lifespan in these animals. PMID- 9853270 TI - Regulation of human renin gene promoter activity: a new negative regulatory region determines the responsiveness to TNF alpha. AB - BACKGROUND: The renin-angiotensin system has been known to regulate blood pressure and body fluid homeostasis. Several lines of evidence have shown that renin gene expression and release are up-regulated by beta-adrenergic stimulation, sodium depletion, and angiotensin converting enzyme inhibition, but down-regulated by cytokines. To further characterize the human renin gene (hREN) promoter structure, its regulation, and to identify an appropriate cell system for study, we examined five cell lines and investigated drug effects on the hREN promoter expression. METHODS: Using the hREN-luciferase reporter gene constructs in the DNA transfection assays, approximately 5 kb of the hREN 5' flanking region was assessed for promoter activity in five different cell lines. Regulation of the hREN promoter activity was investigated using Y-1 adrenal cells that were transfected with the hREN-luciferase DNA and were treated with forskolin, calcium ionophore A23187, phorbol ester, angiotensin II (Ang II), or cytokines. RESULTS: Transient transfection analysis showed that the 5 kb hREN 5' flanking DNA alone was able to confer significant promoter activity in Y-1 adrenal cells. In transfected Y-1 cells, luciferase reporter expression was induced by forskolin, suppressed by the calcium ionophore A23187, and phorbol ester in a dose-dependent manner, but was unaffected by angiotensin II (Ang II). However, when Y-1 reporter cells were transfected with human angiotensin II receptor type 1 (AT1) cDNA, hREN promoter activity was dose-dependently down-regulated by Ang II, which was blockable by losartan, an AT1-selective antagonist. Further studies also showed that hREN promoter activity in Y-1 cells was selectively down-regulated by TNF alpha. Deletion of the hREN promoter sequences between position -3916 and -2822 not only enhanced hREN promoter activity by approximately tenfold, but also caused a failure of down-regulation by TNF alpha. In contrast, neither interleukin (IL)-1 alpha, IL-1 beta, IL-2, nor IL-6 exerted any significant effect. CONCLUSIONS: Together the results suggest that TNF alpha is a negative regulator of the hREN expression in the adrenal cells, and that the TNF alpha responsiveness may be controlled by elements located between the positions -3916 and -2822 of the hREN promoter. Moreover, the Y-1 cell line may provide a valuable model system for studying renin gene regulation. PMID- 9853271 TI - Temporal relationships between hormonal and hemodynamic changes in early human pregnancy. AB - BACKGROUND: The systemic hemodynamic profile of human pregnancy is characterized by a decrease in mean arterial pressure, a rise in cardiac output and plasma volume in association with an increase in renal plasma flow and glomerular filtration rate. The factors and the time course responsible for the initial hemodynamic changes seen in human pregnancy have not been completely documented. We hypothesize that systemic and renal hemodynamic changes occur early, prior to the presence of the fetal-placental unit. METHODS: Thirteen women were studied prior to and immediately following conception in identical fashion at gestational weeks 6, 8, 10, 12, 24 and 36. Individuals underwent mean arterial pressure, cardiac output, inulin and PAH clearance determinations. RESULTS: Mean arterial pressure decreased by six weeks gestation (mid follicular 81.5 +/- 2.6 vs. six weeks 68.7 +/- 2.0 mm tig, P < 0.001) in association with a significant increase in cardiac output, a decrease in systemic vascular resistance and an increase in plasma volume. Renal plasma flow and glomerular filtration rate increased by six weeks gestation. Plasma renin activity and aldosterone concentration increased significantly by six weeks, whereas norepinephrine levels did not change throughout pregnancy. Atrial natriuretic peptide levels increased later, at 12 weeks gestation. Plasma cGMP levels decreased and cGMP clearance increased by six and eight weeks, respectively. CONCLUSIONS: Peripheral vasodilation occurs early in pregnancy prior to full placentation in association with renal vasodilation and activation of the renin-angiotensin-aldosterone system. Plasma volume expansion occurs early, followed later by increases in ANP concentration, suggesting that ANP increases in response to changes in intravasular volume. PMID- 9853272 TI - A simultaneous study of the metabolism of apolipoprotein B and albumin in nephrotic patients. AB - BACKGROUND: The nephrotic syndrome is characterized by proteinuria, hypoalbuminemia and hyperlipidemia. Despite intensive research it is not clear at present what the causal links are between these pathological findings. METHODS: Stable isotope labeled amino acid tracer kinetic analysis was used to simultaneously investigate the metabolism of four apolipoprotein B-containing lipoproteins (VLDL1, VLDL2, IDL and LDL) and albumin in seven patients with nephrotic syndrome and marked hypercholesterolemia, in two additional nephrotic patients with concomitant renal failure and mixed hyperlipidemia, and in a matched group of normolipidemic controls. RESULTS: Increased concentrations of VLDL2, IDL and LDL were due to (a) impaired VLDL2 and IDL delipidation, (b) reduced LDL catabolism, and (c) a trend towards an increased rate of total apolipoprotein B production. The rate of fractional albumin elimination was three times higher in patients than in controls and the rate of albumin synthesis was increased by 45%. No correlations were detectable between rates of apolipoprotein B production and the rate of albumin synthesis. CONCLUSIONS: The results of this study suggest that hyperlipidemia in nephrotic syndrome is predominantly the result of delayed lipoprotein delipidation and catabolism. There is no evidence that it is driven by a general increase of the rate of hepatic protein synthesis. PMID- 9853273 TI - An epidemic of proteinuria in Pima Indians with type 2 diabetes mellitus. AB - BACKGROUND: The risk of proteinuria in Type 1 diabetes declined > or = 30% over the past 50 years, and improvements in metabolic control are believed to be largely responsible. Little is known about secular changes in the risk of proteinuria in Type 2 diabetes. METHODS: We examined trends in the incidence rate of proteinuria in Pima Indians > or = 20 years of age with diabetes diagnosed between January 1, 1955 and December 31, 1994. RESULTS: Among 1305 initially non proteinuric diabetic subjects, 433 developed proteinuria during a median follow up of 8.0 years (range 0.8 to 30.2 years). With subjects with diabetes diagnosed between 1955 and 1964 serving as the reference group, the rate of proteinuria was similar (rate ratio 1.0; 95% confidence interval, 0.79 to 1.3) in the cohort diagnosed between 1965 and 1974, 1.5 times as high (95% confidence interval, 1.1 to 2.0) in the cohort diagnosed between 1975 and 1984, and 1.9 times as high (95% confidence interval, 1.1 to 3.0) in the cohort diagnosed between 1985 and 1994, after adjusting for potential confounders in a generalized additive proportional hazards model. Between the first and last cohorts, plasma glucose concentration declined, on average, by 17% (P = 0.0001) and the mean arterial pressure declined by 11% (P = 0.0001). CONCLUSIONS: The incidence rate of proteinuria in Pima Indians with Type 2 diabetes increased nearly twofold in the last 40 years, despite improvements in plasma glucose and blood pressure. Rapidly changing environmental or behavioral factors must play an important role in the pathogenesis of diabetic renal disease in this population. PMID- 9853274 TI - Effect of indomethacin on the renal response to angiotensin II receptor blockade in healthy subjects. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs are known to promote sodium retention and to blunt the blood pressure lowering effects of several classes of antihypertensive agents including beta-blockers, diuretics and angiotensin converting enzyme (ACE) inhibitors. The purpose of the present study was to investigate the acute and sustained effects of indomethacin on the renal response to the angiotensin II receptor antagonist valsartan and to the ACE inhibitor enalapril. METHODS: Twenty normotensive subjects maintained on fixed sodium intake (100 mmol sodium/day) were randomized to receive for one week: valsartan 80 mg o.d., enalapril 20 mg o.d., valsartan 80 mg o.d. + indomethacin 50 mg bid and enalapril 20 mg o.d. + indomethacin 50 mg bid. This single-blind study was designed as a parallel (valsartan vs. enalapril) and cross-over trial (valsartan or enalapril vs. valsartan + indomethacin or enalapril + indomethacin). Renal hemodynamics and urinary electrolyte excretion were measured for six hours after the first and seventh administration of each treatment regimen. RESULTS: The results show that valsartan and enalapril have comparable renal effects characterized by no change in glomerular filtration rate and significant increases in renal plasma flow and sodium excretion. The valsartan- and enalapril induced renal vasodilation is not significantly blunted by indomethacin. However, indomethacin similarly abolishes the natriuresis induced by the angiotensin II antagonist and the ACE inhibitor. CONCLUSIONS: This observation suggests that although angiotensin receptor antagonists do not affect prostaglandin metabolism, the administration of a non-steroidal anti-inflammatory drug blunts the natriuretic response to angiotensin receptor blockade. PMID- 9853275 TI - Proteinuria selectivity index based upon alpha 2-macroglobulin or IgM is superior to the IgG based index in differentiating glomerular diseases. Technical note. AB - BACKGROUND: The proteinuria selectivity index (SI) may be used to describe changes of the glomerular permeability for macromolecules in glomerular diseases. Proteins the size of alpha 2-macroglobulin (alpha 2 M) or IgM cannot normally pass the glomerular barrier, whereas IgG can pass through the large pores of glomerular basement membrane. Comparison of the clearance of the three high molecular-weight proteins to that of albumin may be useful in characterization and diagnosis of different glomerular diseases as well as in understanding of the permeability characteristics of the glomerular filter. METHODS: Three types of SI, each calculated as a ratio of clearance of either IgG, alpha 2M or IgM to that of albumin, were measured in 199 proteinuric patients. The patients were subdivided into eight different biopsy-verified glomerular diseases. RESULTS: Two diagnoses could be clearly distinguished using SI based on alpha 2M (alpha 2 M SI) or IgM (IgM SI). Both alpha 2M SI and IgM SI were significantly lower in minimal change nephropathy and higher in crescentic necrotizing glomerulonephritis than in all the other diagnoses. The SI based on IgG (IgG SI) was less useful in determining specific diagnoses, since patients with minimal change nephropathy could not be distinguished from those with other types of primary glomerulonephritis and patients with crescentic necrotizing glomerulonephritis did not differ from those with diabetic nephropathy. CONCLUSIONS: The findings of this study indicate that alpha 2M SI and IgM SI are superior to IgG SI in characterization of glomerular disorders and might replace the IgG SI for this purpose. PMID- 9853276 TI - Interleukin-1 receptor antagonist synthesis by peripheral blood mononuclear cells in hemodialysis patients. AB - BACKGROUND: Pro-inflammatory cytokines like interleukin (IL)-1 beta and tumor necrosis factor-alpha (TANF-alpha) are believed to play a significant role in dialysis-related morbidity. It has been previously demonstrated that the endogenous synthesis of interleukin-1 receptor antagonist (IL-1Ra) is a reliable marker of the level of IL-1 beta synthesis in hemodialysis (HD) patients. In this study, we assessed the impact of clinical and laboratory variables on IL-1Ra synthesis by peripheral blood mononuclear cells (PBMC) in patients on HD with unsubstituted cellulose dialyzers. METHODS: IL-1Ra by PBMC was measured by a specific non-cross-reactive radioimmunoassay. Day to day variation in cytokine synthesis, the correlation between cytokine synthesis under different in vitro stimulatory conditions, and the influence of clinical and laboratory variables on cytokine synthesis were studied. RESULTS: Although there was a trend towards greater IL-1Ra synthesis by unstimulated, endotoxin-stimulated and IgG-stimulated PBMC drawn before the second and third dialysis sessions of the week when compared to the first dialysis treatment, this was not statistically significant. There was a strong correlation between IL-1Ra synthesis by PBMC cultured under different stimulatory conditions that was best observed between IL-1Ra cell content and from endotoxin-stimulated PBMC (r = 0.51, P = 0.0001), and endotoxin- and IgG-stimulated PBMC (r = 0.44, P = 0.0001). In addition, there was a close correlation between total synthesis (cell associated and secreted) and secreted levels of IL-1Ra in unstimulated (r = 0.59, P = 0.0001) and endotoxin-stimulated PBMC (r = 0.69, P = 0.0001). Interestingly, there was an inverse correlation between IL-1Ra synthesis and duration of dialysis that was strongest for secreted IL-1Ra from unstimulated (r = -0.50, P = 0.002) and endotoxin-stimulated PBMC (r = -0.34, P = 0.04). There was no significant correlation between IL-1Ra synthesis by PBMC and other clinical and laboratory indices. CONCLUSIONS: The observations from this study indicate that: (1) in HD patients, there were no significant differences in cytokine synthesis by PBMC drawn before the three different dialysis treatments during the week; (2) there is a close relationship between IL 1Ra synthesis from PBMC cultured under different stimulatory conditions; (3) the secreted levels of IL-1Ra correlate directly with total synthesis (cell associated and secreted); (4) with the exception of duration of dialysis, none of the other clinical or laboratory parameters correlated with cytokine synthesis; and (5) the diminished endotoxin- or IgG-stimulated IL-1Ra synthesis with increasing time on dialysis is possibly another sign of the impaired host-defense system in patients on long-term hemodialysis. PMID- 9853277 TI - Inhibition of intercellular adhesion molecule-1 with antisense deoxynucleotides prolongs renal isograft survival in the rat. AB - BACKGROUND: Delayed graft function from ischemia-reperfusion injury has a negative impact on long-term renal graft survival. We tested the utility of antisense oligodeoxynucleotide (ODN) against intercellular adhesion molecule-1 (ICAM-1) in the pretransplant treatment of renal isografts in improving long-term graft survival. METHODS: Three groups of 16 inbred Lewis rats each underwent unilateral nephrectomy and were then transplanted with a kidney from a Lewis donor rat, which had received antisense ODN, reverse sense ODN, or saline vehicle six hours prior to nephrectomy. The kidneys were subjected to one hour of warm ischemia and 30 minutes of cold ischemia, which when untreated results in delayed graft function. The remaining native kidney was removed 10 days later. Serum creatinine and urinary protein excretion were measured in surviving rats at weeks 2, 4, 6, 8, 12, 16, and 20 after native nephrectomy. RESULTS: A Kaplan-Meier analysis revealed that by week 6 one half of the animals receiving reverse sense ODN and saline vehicle treatment had died, while all but 2 rats in the antisense ODN-treatment group survived to 20 weeks. Serum creatinine concentrations and urine protein excretion of surviving reverse sense and saline vehicle-treated rats were significantly higher than antisense treated rats at every time point. Histology at week 20 revealed marked interstitial fibrosis, focal glomerular sclerosis, vascular intimal and medial thickening and tubular atrophy in reverse sense and saline vehicle-treated kidneys, while antisense ODN-treated kidneys showed only modest changes. Immunohistochemistry showed macrophage and lymphocyte infiltration, as well as substantial up-regulation of MHC class II, in reverse sense and saline vehicle-treated kidneys compared to antisense ODN-treated kidneys. CONCLUSIONS: These results suggest that by ameliorating acute nonimmunological renal isograft injury, the long-term chronic nonimmunologic processes are improved as well. Furthermore, the data suggest that an antisense ODN strategy directed against ICAM-1 may have utility in human kidney transplantation. PMID- 9853278 TI - Prevention of peritonitis with disconnect systems in CAPD: a randomized controlled trial. The Mexican Nephrology Collaborative Study Group. AB - BACKGROUND: Recently, disconnect systems for CAPD that are associated with a reduced frequency of peritonitis have been introduced. Our objective was to compare the incidence of peritonitis using three current CAPD systems in a high risk population with low educational and socioeconomic levels, and high prevalence of malnutrition. METHODS: In a prospective controlled trial, 147 patients commencing CAPD were randomly assigned to one of three groups: 29 to the conventional, 57 to the Y-set, and 61 to the twin bag systems. The number of peritonitis episodes was registered, and patients were followed up for an average of 11.3 months. RESULTS: The average peritonitis-free interval for the conventional group was 6.1 months, for the Y system was 12.0 months, and for the twin bag was 24.8 months (P < 0.001). By multivariate analysis, the only factor associated with peritonitis was the CAPD system. Peritonitis-related hospitalization was 5.3 +/- 2.0, 2.7 +/- 1.0, and 1.5 +/- 0.9 days/patient/year in the conventional, Y system, and twin bag groups, respectively. The cost per bag was similar for the conventional and Y system, but higher for the twin bag. However, the total costs of treatment (pesos/patient/year) were lower for twin bag (62,159 for the conventional, 70,275 for the Y system, and 54,387 for the twin bag), due to the lower peritonitis incidence and associated hospitalizations. CONCLUSIONS: Y system and twin bag use was associated with a reduction of 50 and 75% peritonitis incidence, respectively, in patients on CAPD. The cost of the twin bag was actually lower, because of savings from a decreased usage of antibiotics and fewer hospitalizations. PMID- 9853279 TI - Secular trends in recombinant erythropoietin therapy among the U.S. hemodialysis population: 1990-1996. AB - BACKGROUND: Chronic anemia is a major cause of morbidity among the end-stage renal disease (ESRD) population. Recombinant erythropoietin (rHuEPO) has been recognized as a major advance in the treatment of anemia among the ESRD population. This study examines the secular trends in the use of and response to rHuEPO therapy among severely, moderately and mildly anemic hemodialysis patients. METHODS: We designed a cohort analytic study using seven years of claims data. The study population comprised all facility-based adult hemodialysis patients receiving rHuEPO therapy, who were initially reimbursed by Medicare in each of the first quarter of the calendar years 1990 through 1996 (N = 64,957). RESULTS: Between 1990 and 1996, the mean rHuEPO dose increased by 139% for the patient cohorts with a first observed hematocrit < 0.25, 122% for the 0.25 to 0.29 cohorts, and 107% for the > or = 0.30 cohorts, and produced a 0.02 to 0.03 increase in achieved hematocrit (A-Hct) over this time. Dosing of rHuEPO did not appear to be influenced by patient or provider characteristics, although African Americans, the elderly, non-diabetics and persons receiving dialysis in a non profit facility had a larger percent change in hematocrit compared to their counterparts (P < 0.001). CONCLUSIONS: The results of the clinical use of rHuEPO seven years after FDA approval found in the general ESRD hemodialysis population have not equaled the results obtained in the initial clinical trials. Overall, our findings suggest that substantial increases in rHuEPO dose provided to anemic patients have resulted in only modest increases in hematocrit in the seven years since rHuEPO's introduction. Resistance to rHuEPO, prior rHuEPO treatment, inadequate use of supplemental iron, and policy and financial incentives may explain this finding. PMID- 9853280 TI - Effect of serum phosphate on parathyroid hormone secretion during hemodialysis. AB - BACKGROUND: Recent studies have demonstrated that a high concentration of phosphate directly stimulates parathyroid hormone (PTH) secretion. High serum levels of phosphate are usually observed in patients with end-stage renal disease. The aim of the present study was to evaluate whether serum phosphate concentration had an acute effect on PTH secretion in hemodialysis patients. The levels of serum phosphate were manipulated during the hemodialysis session by using a phosphate free dialysate or a dialysate with a high content of phosphate. METHODS: Ten stable hemodialysis patients with PTH values above 300 pg/ml were included in the study. A PTH-calcium curve was obtained during both high phosphate and phosphate free hemodialysis. RESULTS: The serum phosphate concentration remained high (2.17 +/- 0.18 mM) throughout the high phosphate hemodialysis and decreased progressively to normal levels (1.02 +/- 0.06 mM) during the phosphate free hemodialysis. The serum PTH levels at maximal inhibition by hypercalcemia (minimal PTH) were greater during the high phosphate than the phosphate free hemodialysis (413 +/- 79 vs. 318 +/- 76 pg/ml, P < 0.003). In all patients the values of minimum PTH were greater during the high phosphorus than the phosphorus free hemodialysis. The values of maximally stimulated PTH during hypocalcemia and the set point of the PTH-calcium curve were similar during the high phosphate and the phosphate free hemodialysis. CONCLUSION: The maintenance of high serum phosphorus levels during hemodialysis prevented, in part, the inhibition of PTH secretion by calcium, which strongly suggests that in hemodialysis patients high serum phosphate contributes directly to the elevation of PTH levels despite normal or high serum calcium concentration. PMID- 9853281 TI - Abbreviated kinetic profiles in area-under-the-curve monitoring of cyclosporine therapy. Technical note. AB - BACKGROUND: The new microemulsion formulation of cyclosporine (CsA-ME) displays more consistent pharmacokinetic properties than the original formulation and may allow successful implementation of an abbreviated area-under-the-curve (AUC) strategy. METHODS: Here we compared two limited sampling strategies in order to define the one that best predicts AUC after CsA-ME in 51 renal transplant recipients with stable renal function. Pharmacokinetics were based on analysis of blood samples collected over 12 hours after drug administration by high performance liquid chromatography (HPLC). Predicted AUC was estimated by using a three-point (0, 1 and 3 hr) or a two-point (2 and 6 hr or 0 and 2 hr) sampling strategy. RESULTS: A simplified strategy with three time points of blood collection at 0, 1, and 3 hours after CsA-ME allowed adequate and accurate prediction of the daily exposure to CsA. AUC prediction with two-point sampling at 2 and 6 hours was less good with a very large error in prediction (only 59% of the estimated AUC were within the accepted range). This limitation was even more evident when the 0 and 2 hour time points were examined, in which only 51% of AUC estimates were included in the accepted range of variation (-10 to 10%). CONCLUSIONS: A limited strategy of three-point sampling taken early after dosing allows an excellent and perfectly reliable prediction of the actual AUC. PMID- 9853282 TI - Novel roles for chemokines and fibroblasts in interstitial fibrosis. AB - BACKGROUND: Regardless of its involvement in either wound healing or excessive fibrosis, the interstitial fibroblast can now be considered an important early participant in inflammatory responses. Although it is recognized that certain immune cells and proinflammatory mediators are intricately linked to fibrotic disease, little is presently known about the manner in which these mediators and cells are orchestrated to a fibrotic finale. Experimental studies have shown that interstitial fibroblasts are capable of participating in an inflammatory response by promoting direct fibroblast-to-immune cell communication and/or modulating the release of soluble mediators that are mutually recognized by both types of cells. METHODS: Primary cultures of murine fibroblasts were recovered from either normal tissue or tissue undergoing a cell-mediated inflammatory response. These stromal cells were assessed for the expression of various cytokines and chemokines indicative of a type 1 or type 2 response. In addition, the fibroblasts were co cultured with mononuclear cells to assess the cell-to-cell communication. RESULTS: Fibroblasts recovered from different cell-mediated inflammatory responses demonstrated a dramatic alteration in their cytokine profile. Fibroblasts recovered from the type 2 immune response produced high levels of monocyte chemotactic protein-1 (MCP-1), as compared to the normal fibroblasts and fibroblasts recovered from the type 1 lesion. Mononuclear cells co-cultured with fibroblasts induced a contact-dependent expression of elevated levels of chemokines, especially the macrophage-derived MIP-1 alpha. Thus, both fibroblasts themselves and fibroblasts co-cultured with immune-inflammatory cells have the ability to participate in the maintenance of an inflammatory response via the expression of chemokines. CONCLUSIONS: Our laboratory and others have addressed the role of chemotactic cytokines or chemokines in the fibrotic process, and have demonstrated that fibroblasts are capable of modulating the activation of various immune cells that have been implicated in fibrotic disease. In addition, the interstitial fibroblast is capable of regulating its own behavior within the interstitial environment via the expression of chemokines and chemokine receptors. Thus, novel strategies aimed at preventing fibrotic disease will likely need to address the early engagement of inflammatory cells by fibroblasts, and possibly modulate the ability of fibroblasts to generate and/or recognize profibrotic signals supplied by chemokines. PMID- 9853283 TI - Response of the human peritoneal mesothelial cell to injury: an in vitro model of peritoneal wound healing. AB - BACKGROUND: The denudation of the peritoneal mesothelium and damage to the underlying interstitium is a frequent finding in patients receiving continuous ambulatory peritoneal dialysis as a treatment for end-stage renal failure. The response of the mesothelium to injury from repeated episodes of infection or from exposure to dialysis fluids has not been extensively studied. The present study describes a simple and reproducible method with which to investigate the response of human mesothelial cells to injury. METHODS: The model of peritoneal injury consists of mechanically wounding a monolayer of human peritoneal mesothelial cells with a glass probe and following the repopulation of the denuded area by time-lapse photomicroscopy. In addition immunohistochemistry was used to follow the response of marker proteins for stress fibers and focal adhesions as well as macromolecules associated with the extracellular matrix. RESULTS: Under serum free conditions the wound (0.58 +/- 0.094 mm; mean +/- SD; N = 20) closed within 72 +/- 5 hours (N = 8). This rate of healing was enhanced by fetal calf serum, by human serum (10%) and by undiluted spent non-infected dialysate. The repair process over the first 48 hours was the result of cell migration, was independent of cell proliferation and involved the de novo synthesis of several different extracellular matrix components. An early event in the healing process was the rapid reorganization of intracellular stress fibers together with the formation of associated focal adhesions in cells at the wound edge. CONCLUSION: This in vitro model should prove invaluable in characterizing the process of wound healing within the peritoneal cavity, thus allowing a better understanding of the response to infection as well as any effect of dialysis fluids in this pattern of cell behavior. PMID- 9853284 TI - Leukocyte migration across human peritoneal mesothelial cells is dependent on directed chemokine secretion and ICAM-1 expression. AB - BACKGROUND: Leukocyte migration into the peritoneal cavity is a diagnostic feature of peritonitis in patients treated with peritoneal dialysis (PD). While neutrophil (PMN) influx is characteristic of the acute phase of peritoneal infection, significant mononuclear cell (MNC) infiltration, occurs throughout the whole period of infection. Recent data suggests that human peritoneal mesothelial cell (HPMC) adhesion molecule expression and the synthesis of chemotactic cytokines may be important in the process. METHODS: In the present study we have examined, the regulation and directed secretion of chemokines (IL-8, MCP-1 and RANTES) and the basolateral to apical migration of unstimulated leukocytes across mesothelial cell monolayers using an in vitro model where HPMC were grown on the porous membrane of tissue culture inserts. Separate experiments have defined the importance of chemokine synthesis and ICAM-1 expression in the transmigration process. RESULTS: Apical stimulation of HPMC with IL-1 beta or TNF alpha resulted in a time and dose dependent up-regulation of IL-8, MCP-1 and RANTES mRNA expression and synthesis. This secretion was predominately into the apical compartment (> 85%) with all chemokines. Apical pre-stimulation of HPMC resulted in a dose- and time-dependent migration of both PMN and MNC across HPMC. Neutrophil migration was significantly reduced in the presence of appropriate concentrations of polyclonal IL-8 antibody (IL-1 beta (100 pg/ml) 153 +/- 12 versus anti-IL-8 (100 ng/ml) 71 +/- 7 (X 10(3)) PMN, N = 6, P < 0.02) and in the presence of anti-ICAM-1 F(ab)'2 fragments or soluble ICAM-1. Constitutive and cytokine stimulated mononuclear cell migration was significantly reduced in the simultaneous presence of polyclonal MCP-1 or RANTES antibody. CONCLUSIONS: These data demonstrate that HPMC synthesize IL-8, MCP-1 and RANTES in response to inflammatory cytokines. HPMC-derived C-x-C and C-C chemokines might contribute to the intra-peritoneal recruitment of leukocytes during peritoneal inflammation. PMID- 9853285 TI - Biocompatibility and buffers: effect of bicarbonate-buffered peritoneal dialysis fluids on peritoneal cell function. AB - BACKGROUND: Conventional peritoneal dialysis fluids (PDF) have been shown to compromise the function of both leukocytes and human peritoneal mesothelial cells (HPMC). Various in vitro studies have identified the low initial pH in combination with high lactate content, as well as the hyperosmolality and high glucose concentration present in currently used solutions as the primary determinants of their bioincompatibility. Bicarbonate buffered PDF (at neutral pH) display improved in vitro biocompatibility as compared to conventional, lactate buffered PDF. However, little information is currently available regarding the potential impact of PDF on the function of human peritoneal fibroblasts (HPFB), the major cell population present in peritoneal interstitium. METHODS: The current study compares the effect of bicarbonate and lactate buffered PDF in a model system of resting peritoneal mesothelial cells and fibroblasts cultured from human omentum. Interleukin-1 beta-stimulated IL-6 release from HPMC and HPFB was used as the cell functional parameter. RESULTS: While short (30 min) pre-exposure to lactate buffered PDF significantly reduced the IL-1 beta-stimulated IL-6 release from HPMC during a subsequent recovery period (24 hr), a significant decrease in HPMC IL-6 secretion with bicarbonate buffered PDF was only observed after prolonged (> or = 60 min) exposure. In contrast, no significant IL-6 inhibition was detected with HPFB pre-exposed to PDF for up to 90 minutes. A significant suppression of HPFB IL-6 secretion was only observed in coincubation experiments (24 hr) with dilutions of both types of PDF. CONCLUSIONS: These results indicate that (i) bicarbonate buffered PDF are less inhibitory to peritoneal cell function as compared to conventional, lactate buffered PDF; and (ii) HPFB may be more resistant than HPMC to bioincompatible PDF. PMID- 9853286 TI - Experimental models in peritoneal dialysis: a European experience. AB - BACKGROUND: The development of adequate animal models is important for the in vivo study of selected aspects of peritoneal dialysis (PD) that cannot be evaluated by an in vitro model, such as peritoneal membrane transport, the influence of local defense mechanisms, and for testing new osmotic agents and their biocompatibilities. METHODS: Our experience with animal models for PD, including the acute Stockholm model in non-uremic rats, the acute and chronic Amsterdam model in non-uremic rats, and the chronic Gent model in uremic rats, is described. RESULTS: The Stockholm model proved to be useful in understanding the normal physiology of peritoneal transport, and for testing new dialysis solutions and their biocompatibilities. It is a rather simple and inexpensive model, and thus is suitable for screening new solutions and additives. The Amsterdam model permits the study of chemokines and mesothelial cell regeneration in vivo, and is applied in a model of chronic peritonitis. The results of the Gent model suggest that chronic peritoneal dialysis in uremic rats is feasible for at least eight weeks. This model is, however, very laborious, time consuming, and expensive. CONCLUSION: Further improvement of the technique and increase of the dialysis dose should result in a better and more realistic model for peritoneal dialysis. It is hoped that in the future these models will be useful to test the effects of long-term intraperitoneal application of different dialysis solutions and additives in uremic animals. PMID- 9853287 TI - What really happens to people on long-term peritoneal dialysis? AB - BACKGROUND: Several risk factors for patients treated with peritoneal dialysis (PD) have now been identified. These include age, comorbid disease, nutritional status, loss of residual renal function (RRF) and high peritoneal solute transport. This is not the same, however, as knowing what actually happens to these patients, particularly in the long-term. The purpose of this review was to give as complete a description as is currently possible of the long-term PD patient. METHODS: The literature was surveyed for publications that provide longitudinal cohort data of either selected or unselected patient groups. Detailed data from the Stoke PD Study is presented in the context of these studies. Three principle aspects of what really happens to patients were considered: (1) death, both cause and mode of death; (2) technique failure, with reference to peritoneal function and how the cause of technique failure related to patient survival; and (3) evolution of clinically relevant parameters of patients on PD, such as nutrition and peritoneal function. RESULTS: Sudden death and debilitation were the predominant modes of death, with sepsis playing a contributory role. Debilitation was important regardless of co-existent comorbid disease, and time to death was not influenced by the mode of death. Predominant causes for technique failure remain peritonitis and ultrafiltration, the latter becoming more important with time on treatment. Technical failure is associated with poorer survival, particularly when due to multiple peritonitis or failure to cope with treatment. Cox regression demonstrated that whereas low albumin, loss of RRF and high solute transport predicted patient death, only high solute transport predicted technique failure. Longitudinal changes over the first five years of treatment included loss of RRF, increasing solute transport and following an initial improvement in nutritional state, a decline after two years. Patients surviving long-term PD (at least five years, N = 25) were characterized by prolonged RRF, maintained nutrition and lower solute transport in the medium term. CONCLUSIONS: Several studies of long-term PD in the literature now complement each other in providing a picture of what really happens to PD patients. The links between loss of solute clearance and poor peritoneal ultrafiltration combining to exacerbate sudden or debilitated death and technique failure are emerging. For PD to be successful as a long-term therapy, strategies that maintain nutrition and preserve peritoneal membrane function must be developed. PMID- 9853289 TI - Prevention of renal osteodystrophy in peritoneal dialysis. AB - BACKGROUND: Renal osteodystrophy (ROD) is still one of the major long-term complications in end-stage renal disease leading to considerable morbidity. Despite some progress in understanding the pathogenesis of secondary hyperparathyroidism (sHPT) during recent years, prevention and treatment of ROD is still suboptimal, requiring surgical parathyroidectomy in 6 to 10% of all patients on dialysis after 10 years. In addition, the spectrum of bone lesions has changed, with non-aluminum-related adynamic bone disease (ABD) found in up to 43% of peritoneal dialysis (PD) patients. METHODS: Current recommendations concerning prevention of ROD in PD based on the literature and personal recent data were reviewed. The focus is on (i) the importance of early prophylactic intervention to prevent parathyroid gland hyperplasia, (ii) the pathogenesis of ABD, and (iii) the role of metabolic acidosis in ROD. RESULTS: There is ample evidence that sHPT starts early during the course of renal failure and results from both hypersecretion of PTH by parathyroid cells and glandular hyperplasia. As shown by experimental and clinical studies, established parathyroid cell hyperplasia is hardly reversible by pharmacological means, and therefore prevention of parathyroid cell proliferation needs to start early. Recent data from randomized trials document the efficacy and safety of low dose active vitamin D (0.125 to 0.25 microgram/day) and/or an oral calcium substitute to prevent progression of sHPT in patients with mild to moderate renal failure. Since little is known about the pathogenesis, natural course and clinical impact of ABD in PD, specific therapeutic concepts have not yet been generated. Diabetes and advanced age are established risk factors, whereas the role of calcium and vitamin D overtreatment or the type of dialysis (PD vs. HD) are still controversial. Currently no evidence for different functional behavior of the parathyroids in ABD and sHPT has been found. The role of circulating or local factors such as cytokines, growth factors or the presence of advanced glycation end-product (AGE)-modified matrix proteins for the pathogenesis of either type of ROD deserves further investigation. Avoiding oversuppression of parathyroid gland and the use of low calcium dialysate may help prevent ABD. There is growing evidence that a correction of metabolic acidosis will influence ROD by both direct effects on the bone and on parathyroid cell function. New dialysate composition for CAPD with a high HCO3 concentration will allow normalization of acid-based metabolism in PD patients. Their effects on ROD under long term conditions remain to be determined. CONCLUSION: Therapeutic efforts should aim to prevent the development of parathyroid gland hyperplasia and sHPT early during the course of renal failure, and should include the use of low dose vitamin D therapy and oral calcium substitution as well as correction of metabolic acidosis. Concerning ABD, more information is needed regarding the causes and consequences of this type of bone lesion to develop a more specific therapy. PMID- 9853288 TI - Augmenting solute clearance in peritoneal dialysis. AB - BACKGROUND: The removal of low molecular weight solutes by peritoneal dialysis is less than by hemodialysis. The targets for Kt/Vurea and creatinine clearance formulated in the Dialysis Outcome Quality Initiative are unlikely to be achieved in a substantial portion of peritoneal dialysis patients. Possibilities to increase small solute clearances have therefore been subject to many investigations. METHODS: A review of the literature and of recent new data on determinants of solute removal, such as residual renal function, the role of drained dialysate volume and manipulation of the diffusive capacity of the peritoneum are presented. RESULTS: The contribution of residual GFR is more important for the clearance of creatinine than for Kt/Vurea. It is even more important for the removal of organic acids that are removed from the body by tubular secretion. High dosages of furosemide increase the urinary volume and the fractional Na+ excretion, but have no effect on the magnitude of residual GFR, renal creatinine clearance, renal urea clearance, and peritoneal transport characteristics. The drained dialysate volume per day is the main determinant of the peritoneal removal of urea. Its effect decreases the higher the molecular weight of a solute. It can be augmented by using large instillation volumes, by the application of more exchanges, and by increasing peritoneal ultrafiltration. A large exchange volume is especially effective in patients with an average transport state, but in those with high solute transport rates, Kt/Vurea is especially influenced by the number of exchanges. Possibilities to increase ultrafiltration are discussed. The diffusive capacity of the peritoneum can be augmented by using low dosages of intraperitoneally administered nitroprusside. This increases solute transport most markedly when it is applied in combination with icodextrin as osmotic agent. CONCLUSIONS: Small solutes clearances cannot be increased by furosemide. Increasing the instilled volume of dialysis fluid and the number of exchanges both affect solute clearance. Studies are necessary on long-term effects of manipulation of the peritoneal membrane with nitroprusside. PMID- 9853290 TI - What is the place of peritoneal dialysis in the integrated treatment of renal failure? AB - The role of peritoneal dialysis (PD) in renal replacement therapy (RRT) remains unclear. There are no controlled trials to provide hard evidence of its efficacy. Comparative studies with haemodialysis from different centres and countries have given conflicting results even when allowing for case mix. Data from the United States on patients starting or receiving treatment in the late 1980s suggested a worse prognosis for older patients, particularly diabetics receiving PD as compared to HD. Analysis of the USRDS data base for patients starting in the early 1990s shows an improvement in outcome but with no difference in overall mortality. The Canadian registry has recently published data showing a better survival with PD than with HD in the first two years of RRT. Morbidity is similar with both therapies, although hospitalization is increased with PD. Unfortunately long-term technique survival is not as good with PD. However, PD has certain medical advantages, particularly the maintenance of residual renal function that contributes to solute and fluid removal. It may also postpone the onset of amyloidosis. Patients transplanted after previous PD have a decreased risk of early acute renal failure and equally good long-term results when compared to those patients who were on HD before transplantation. The quality of life is as good with PD as with center HD, and there are social advantages to PD including an increased chance of employment, more flexible holidays and avoidance of thrice weekly travel to a dialysis center. PD also has logistical advantages and can be utilized by the majority of new patients. We therefore conclude that PD has potential advantages early in the course of RRT, and should therefore be offered as a first option to all suitable new patients. Whether PD has a major or minor role in later years (> 5) remains unclear. PMID- 9853291 TI - The growing role for the cyclin kinase inhibitor p27Kip1 in renal disease. PMID- 9853292 TI - Potential role for G-protein coupled receptors in HIV-associated nephropathy. PMID- 9853293 TI - Macrophage migration inhibitory factor in IgA nephropathy. PMID- 9853294 TI - The aging kidney. PMID- 9853295 TI - Effects of a triple enzyme digestion method on a diamine reaction for glycosaminoglycans of the rat aorta in electron microscopy. AB - Effects of a triple enzyme digestion method using glycosaminoglycans-degrading enzymes upon a diamine reaction have been tested for electron microscopic histochemical detection of glycosaminoglycans in extracellular matrix of the rat aorta. The triple enzyme digestion method consists of a sequence of chondroitinase B, testicular hyaluronidase and heparitinase. The results obtained by the present experimental and control studies indicated that dermatan sulfates, chondroitin sulfates (A and/or C) or heparan sulfates were apparently observed in various ultrastructural features of aortic extracellular matrix, such as bundle of collagen fibers and soluble matrix of interstitial space. Particularly, we found that both heparan sulfates and chondroitin sulfates (A and/or C) were detected in association with the basal lamina of smooth muscle cells and the external surface of elastic lamina, and in the latter heparan sulfates were frequently recognized as a mass, whereas chondroitin sulfates (A and/or C) were found intermittently along the external surface of elastic lamina. This suggests that the triple enzyme digestion method which combines the glycosaminoglycans degrading enzymes with the diamine reaction can be postulated to represent efficient and useful technique for precise electron microscopic histochemical detection of the glycosaminoglycans in the extracellular matrix of the rat aorta. PMID- 9853296 TI - Expression pattern of mitochondrial capsule selenoprotein mRNA in the hamster testis. AB - Mitochondrial capsule selenoprotein (MCS) has been known as a structural protein of the mitochondrial sheath in spermatozoa. In the present study, to determine the expression pattern of MCS mRNA in the hamster testis, northern blot and in situ hybridization analyses using digoxigenin-labeled riboprobes for the hamster MCS were performed in the testes of 10-week-old golden hamsters. According to the northern blot analysis, hamster MCS was detected as a single transcript of about 1 kb in the testis. During spermatogenesis, the hamster MCS mRNA first appeared in step 6 spermatids, gradually increased in round spermatids during spermiogenesis, reached a peak in step 8 spermatids, and persisted a high level until step 13 spermatids. After step 14, the signal began to show a progressive decline in the spermatids and was weakly detected in the tails of step 17 spermatids. However, the signal was not observed in spermatogonia, spermatocytes, Sertoli cells, peritubular myoid cells, and interstitial cells. These findings indicate that hamster MCS is mainly related to the spermiogenesis during spermatogenesis. PMID- 9853297 TI - Effect of interleukin-12 and interleukin-10 on the virus replication and apoptosis in T-cells infected with feline immunodeficiency virus. AB - Interleukin-12 (IL-12) is an important cytokine for Th1 response which stimulates the T-cell population to produce cytokines for cellular immunity. Interleukin-10 (IL-10) is a pleiotropic cytokine capable of suppressing cytokine production from macrophages and T-cells and participants in Th2 immune response. The present study was carried out to examine the effect of these cytokines on virus replication and apoptosis in T-cells infected with feline immunodeficiency virus (FIV). Infection of a feline T-lymphoid cell line (Fel-039) resulted in an increase of the reverse transcriptase (RT) activity in the culture supernatant accompanied by cell death from apoptosis. Addition of human recombinant IL-12 significantly inhibited the virus replication and apoptosis in Fel-039 cells in a dose dependent manner. Furthermore, the antiviral activity of IL-12 was associated with the expression of IFN-gamma in the FIV-infected Fel-039 cells. In contrast, human recombinant IL-10 did not show any inhibitory effect on the virus replication and apoptosis in the Fel-039 cells infected with FIV. These results suggest that the inhibitory effect of IL-12 on both virus replication and apoptosis has potential implications for the design of immunotherapy strategies using IL-12 in FIV infection. PMID- 9853298 TI - Inhibition of apoptosis and virus replication in feline immunodeficiency virus infected cells by N-acetylcysteine and ascorbic acid. AB - Infection of feline immunodeficiency virus (FIV) has been shown to induce apoptosis that might be associated with the lymphocyte depletion in the infected cats. To investigate the inhibitory effect of antioxidants on FIV-induced apoptosis, we examined the effect of N-acetylcysteine (NAC) and ascorbic acid (AA) on apoptosis and virus replication in feline lymphoblastoid (Fel-039) and fibroblastoid (CRFK) cell lines infected with FIV. The treatment with NAC or AA induced a significant inhibition of viral replication and apoptosis in Fel-039 cells and tumor necrosis factor alpha (TNF-alpha)-treated CRFK cells infected with FIV. Both cell lines in the presence of noncytotoxic concentrations of NAC or AA showed in increase of intracellular glutathione (GSH) level, which might protect the cells against oxidative stresses exerted by FIV infection and TNF alpha treatment. On the basis of these in vitro results, we suggest that antioxidant therapies aimed at restoring depleted GSH level might be effective for inhibition of viral replication and cell death associated with the development of immunodeficiency. PMID- 9853299 TI - Seroprevalence and field isolation of bovine immunodeficiency virus. AB - A seroprevalence study of bovine lentivirus, known as bovine immunodeficiency virus (BIV), was conducted in 12 different dairy herds in Hokkaido, where some herds were a high prevalence of bovine leukemia virus (BLV) infection. Amongst 611 cattle, 28.6% of cattle were BLV-seropositive, and 11.7% of cattle were seropositive for BIV, while 4.2% of cattle were seropositive for both BIV and BLV. For the isolation of BIV, 19 samples of peripheral blood mononuclear cells (PBMC) and one sample of milk-derived leukocytes were prepared from BIV seropositive cows. These PBMC and leukocyte preparations were then co-cultivated with cc81 cells, a cat cell line transformed by mouse sarcoma virus. BIV was isolated from 17 PBMC and one milk-derived leukocyte samples. The isolated viruses showed slow replication and syncytia formation. Major core antigen, p26 from these isolates were reacted with anti-BIV (American isolate R-29) serum. In addition, proviral DNA was detected in blood and milk samples by nested polymerase chain reaction and subsequent Southern blot hybridization. Nucleotide sequence analysis of the amplified pol gene products showed its 99.0 to 99.7% homology to that of BIV R-29. These results indicate that the Japanese BIV isolates appear to be antigenically and genetically similar to the American R-29. Since BIV was isolated from milk samples, BIV could possibly be transmitted through milk. This is the first report of BIV isolation in Japan. PMID- 9853300 TI - Production of recombinant porcine tumor necrosis factor alpha in a baculovirus expression system. AB - Porcine tumor necrosis factor alpha (TNF-alpha) was produced using a baculovirus system in Spodoptera frugiperda (SF21AE) cells. Cytotoxic activity was detected in supernatant of sonicated SF21AE cells infected with recombinant viruses. The recombinant protein was also demonstrated to be functionally active by its ability to cause apoptosis in Wehi 164 cells. Three distinct bands of 26, 17 and 14 kDa were revealed by Western blot analysis using anti-human TNF-alpha antibody. Moreover, the anti-human TNF-alpha antiserum significantly neutralized the cytotoxic activity of the supernatant of sonicated SF21AE cells infected with recombinant viruses. PMID- 9853301 TI - Molecular identification of a recent type of canine distemper virus in Japan by restriction fragment length polymorphism. AB - Restriction fragment length polymorphism analysis was used to differentiate recent field viruses of canine distemper virus (CDU) from vaccine strains. Virus genomes were amplified by using reverse transcriptase-polymerase chain reaction in part of the haemagglutinin gene. After digestion with EcoRV, the PCR products of recent field isolates were cut into two fragments that differ from the uncut form of old strains including all of vaccine strains. This method could be applied to fresh or stored brains, spleens and peripheral blood mononuclear cells of infected dogs. This molecular approach is useful for determining the causative agent of postvaccinated CDV infection. PMID- 9853302 TI - Enzyme-linked immunosorbent assay for serum procollagen type III peptide in rats with hepatic fibrosis. AB - The process of hepatic fibrosis, and the changes in contents of hepatic hyproxyproline (HYP) and serum procollagen type III peptide (PIIINP) were examined in two rat models for hepatic fibrosis, i.e. bile duct ligation/scission (BDL/s)- and dimethylnitrosamine (DMN)-induced models. In addition, an expression of type III collagen mRNA in the liver of BDL/s model was also examined. In BDL/s model, hepatic fibrosis started at 2 weeks after operation (WAO) and cirrhosis with prominent bile duct hyperplasia was detected at and after 5 WAO. Serum PIIINP content measured using a modified double armed inhibition enzyme-linked immunosorbent assay (ELISA) method proposed by us started to increase at 1 WAO and continued to increase thereafter. Hepatic HYP content measured colorimetrically started to increase at 3 WAO and it continued to increase until 7 WAO. An expression of type III collagen mRNA in the liver was enhanced at and after 2 WAO, especially at 4 and 5 WAO. In DMN model, marked hepatic fibrosis was detected at 1 week after the last DMN administration (WAA), and the degree of fibrosis was apparently reduced at 4 WAA. Serum PIIINP content prominently increased at 1 WAA and decreased at and after 3 WAA. Hepatic HYP content showed a marked increase at 1 WAA and decreased thereafter. The present results indicated that the sequences of hepatic fibrosis, hepatic HYP content and serum PIIINP content were well correlated with each other in both BDL/s and DMN models. In conclusion, ELISA system for the detection of serum PIINP content is considered to be reliable method for assessment of cirrhotic liver, and the present two rat models for liver fibrosis/cirrhosis seems to be a good tool for researching antifibrotic agents. PMID- 9853303 TI - The application of electrocecography for evaluation of cecum motility in horses. AB - Electrogastrography (EGG), in which the electrical activity of the smooth muscular layer of the stomach is recorded percutaneously through the abdominal wall, has been applied in recent years to humans as a non-invasive method. In acute abdominal disease in horses, it is considered diagnostically useful to analyze digestive activity using EGG. Electrocecography (ECG) was examined to determine its effectiveness in evaluating equine digestive motility through comparison, after xylazine administration, between the results of the percutaneous ECG method and the results obtained using a strain-gauge force transducer (Force Transducer) chronically attached to the serous membrane of the cecum. As subjects, the test used six male thoroughbreds (average weight: 457.5 +/- 9.2 kg). The test showed a reduction in both the percutaneous electrical potential of the cecum in ECG and in cecal contractions measured with the Force Transducer. After xylazine administration, an average rates of decrease of the amplitude from the control period were 17.8 +/- 3.4% and 20.0 +/- 4.6% respectively, demonstrating a significant correlation (r = 0.90) between the two methods. On the other hand, power distribution centered around 6 cycles per minute in a Fourier transform (FFT) analysis of ECG, thought similar to the contraction frequency of 5.4 +/- 3.0 per minute observed with the Force Transducer. After xylazine administration, the total frequency band (1.8-12 cycle per min) in the running spectrum total power in ECG decreased to 37.0 +/- 5.1% of the pre-xylazine value. Based on these findings, it appears that the ECG potential reflected electrical activity of cecal origin, suggesting high clinical applicability of ECG to the percutaneous evaluation of equine cecal motility. PMID- 9853304 TI - Potentiation by neurotensin of carbachol-induced tension development in beta escin-skinned smooth muscle of guinea-pig ileum. AB - Effect of neurotensin (NT) on carbachol(CCh)-induced tension development due to Ca2+ release from intracellular stores was investigated in beta-escin-skinned smooth muscle of guinea-pig ileum. NT (10 nM) increased the tension development in response to CCh. NT also increased the tension response to caffeine, another store-Ca2+ releaser. NT did not exert such an effect in pertusis toxin (PTX) treated preparations. Treatment with isoprenaline to elevate endogenous cyclic AMP levels or with dibutyryl cyclic AMP did not affect the effect of NT. A nonpeptide NT antagonist, SR 48692, failed to block the effect of NT. NT shifted the pCa-tension relationship in the lower direction of Ca2+ concentration. NT was incapable of releasing Ca2+ from intracellular stores. The results suggest that NT may cause an increase in Ca2+ sensitivity of contractile elements to potentiate the CCh-induced tension development due to release of stored Ca2+ and that the effect is mediated by SR 48692-insensitive NT receptors linked to a PTX sensitive G protein which works with no relation to a change in cytosolic cyclic AMP levels. PMID- 9853305 TI - Salmonella in broiler chickens in Thailand with special reference to contamination of retail meat with Salmonella enteritidis. AB - The present study was performed to assess the prevalence of Salmonella in chickens in Thailand. In 1997, 22 serovars of Salmonella were isolated from 72 of 100 chicken meat samples purchased from 10 retail markets in Bangkok and 20 of 200 chicken meat samples from one slaughterhouse for export and 19 of 285 chicken feces obtained from three farms located in the east region of Thailand. The most predominant serovar was S. Enteritidis, which was isolated from 28% of the retail chicken meat, 4.5% of the chicken meat from slaughterhouse, and 6.6% of chickens feces samples examined. PMID- 9853306 TI - Circulatory arrest under hypothermic anesthesia using abdominal cavity cooling. AB - The purpose of this study was to investigate the potential for circulatory arrest during surgery under systemic hypothermic anesthesia, using the abdominal cavity cooling method. Eighteen beagles, each weighing 10.5 +/- 2.3 kg, were cooled by filling the abdominal cavity with crushed ice. Just after the esophageal temperature reached 30 degrees C, the heart was exposed, and a left-heart bypass from the left atrium to the aortic root was created. At 20-23 degrees C, the heart was arrested by infusing cooled Young's solution into the aortic root. After a period of cardiac arrest, resuscitation and rewarming were initiated simultaneously. Throughout these procedures, an electrocardiogram (ECG) and the arterial blood pressure (ABP) were monitored continuously. Hematocrit (Ht), total protein (TP), and arterial blood pH and gases were measured every 30 min. The recoveries after surgery were divided into three types as follows, 1) recovery without any complications-11 dogs, 2) not extubated with spontaneous breathing-4 dogs, 3) no reappearance of heart beat-3 dogs. PaO2 during resuscitation was significantly higher in dogs which recovered completely than in the rest of the dogs. These results suggest that hypothermia induced by the abdominal cavity cooling method could be useful for organ-protection during open-heart surgery, and that successful recovery may be attained through protection of the lung as well as the myocardium. PMID- 9853307 TI - Nitroblue tetrazolium reduction of neutrophils in heat stressed goats is not influenced by selenium and vitamin E injection. AB - Experiment was designed to determine whether heat stress suppresses neutrophil function and injections of selenium and vitamin E prior to heat stress prevent suppression of neutrophil function in goats. Twelve female goats were divided into 2 groups of 6 each and were kept at 25 degrees C. Goats in the treatment group were injected intramuscularly with 0.1 mg/kg of selenium and 2.72 IU/kg of vitamin E at 8 and 1 day prior to the initiation of heat stress. The other group was kept as control. All goats were exposed to hot environment at 38 degrees C from day 0 through 8. Decreased tendency in plasma cortisol concentrations and temporary increase in plasma glucose concentrations were shown in both groups. In the control group, plasma selenium concentration gradually increased and alpha tocopherol concentration decreased during the first 2 days. After the second injection with selenium and vitamin E, plasma selenium and alpha-tocopherol concentration significantly increased and remained higher than those in the control group. Whole blood glutathione peroxidase (GSH-Px) activity in the treatment group tended to be greater than that in the control group, but no significant difference was observed between 2 groups. The nitroblue tetrazolium (NBT) reduction by activated neutrophils significantly decreased on day 6 in the control group but not in the treatment group. The NBT reduction by resting neutrophils significantly decreased in both groups. These data suggest that heat stress depresses neutrophil function, and selenium and vitamin E injection prior to heat stress has no apparent effect on neutrophil function during the stress. PMID- 9853308 TI - Metastatic intracavitary cardiac aortic body tumor in a dog. AB - A mobile right-ventricular mass dynamically occluding the right ostium atrioventriculare in the systolic phase was detected in a 3-year-old male Tosa dog by echocardiography. At necropsy, multiple tumor masses of various sizes were observed in the heart base right ventricular lumen, myocardium, lung and liver. Dysplasia of tricuspid valve characterized by irregular shape of leaflets, upward malposition of large papillary muscles, and shortened and stout chordae tendineae was also detected. Histopathologically, the tumor cells, arranged in sheets or nests, were polyhedral with lightly eosinophilic and finely granular cytoplasm, and contained a hyperchromatic round or oval nucleus. By Grimelius' silver stain, tumor cells had cytoplasmic positive granules. Ultrastructurally, tumor cells contained characteristic small membrane-limited granules. This is the first report of metastatic intracavitary cardiac aortic body tumor in a dog. PMID- 9853309 TI - Reactivity of native conglutinin in bovine serum with rabbit antibody against recombinant bovine conglutinin with deletion of the N-terminal and collagen-like regions. AB - The reactivity of native bovine conglutinin (Kg) with antibody against recombinant Kg (rKg), with deletion of the N-terminal and collagen-like regions of the native Kg molecule, was studied by sandwich enzyme-linked immunosorbent assay. With anti-recombinant Kg antibody as the coating antibody, rKg reacted with biotinylated homologous anti-rKg and heterologous anti-Kg antibodies as probing antibodies, while native Kg did not. With anti-native Kg antibody as coating antibody, native Kg reacted with biotinylated homologous antibody as probing antibody, while recombinant Kg reacted weakly with both biotinylated homologous and heterologous antibodies. Consequently the N-terminal and collagen like regions of native Kg molecule are essential to express the complete immunogenicity and/or antigenicity of the native Kg molecule. PMID- 9853310 TI - A study of metabolites isolated from the urine samples of cats and dogs administered orbifloxacin. AB - Urine samples of cats and dogs collected for 24 hr after a subcutaneous injection of orbifloxacin (OBFX) were analyzed. The metabolites were examined using HPLC. In the dog urine, 87% of total was the parent compound and 13% glucuronide compound of OBFX and 96% was parent and 4% metabolite in the cat urine. The metabolite of cat urine was identified as N-hydroxy OBFX, determined by comparison of the extraction of urine with chloroform with the standard compound of N-hydroxy OBFX, using LC/APCIMS. N-hydroxy OBFX had a weaker antibacterial activity against fluoroquinolone sensitive bacteria than the parent compound. PMID- 9853311 TI - A rare case of patent ductus arteriosus in a dog with glomerular mesangiolysis. AB - Rare case of patent ductus arteriosus (PDA) was observed in a 2-year and 9 month old Miniature Dachshund which had been diagnosed as severe cardiac failure. Pulmonary artery from the right ventricle connected to dilated ductus arteriosus, and pulmonary artery-ductus arteriosus-descending aorta formed a continuous duct, which seemed to be the main route of bloodstream. Ascending aorta from the left ventricle was hypoplastic, and connected to the dilated ductus arteriosus. Glomerular mesangiolysis due to heart failure was also observed in the kidney. PMID- 9853312 TI - Relationship of the blood endotoxin concentration and prognosis in dogs with pyometra. AB - The blood concentrations of endotoxin in dogs with pyometra (n = 45) were compared with those in healthy dogs (n = 17). The blood endotoxin concentrations in the healthy dogs (n = 17), in those with good prognosis (n = 41) and those with poor prognosis (n = 4) were 3.4 +/- 2.8 pg/ml, 9.5 +/- 11.3 pg/ml and 74.2 +/- 18.3 pg/ml, respectively. The concentrations in the dogs with good prognosis and poor prognosis were significantly (p < 0.01) higher than those in the healthy dogs. The dogs with poor prognosis had significantly (p < 0.01) higher endotoxin concentrations than those with good prognosis. Blood endotoxin concentrations were measured in 9 dogs after surgery, and were found to be decreased. These results suggest the possible involvement of endotoxin in the pathophysiological changes due to pyometra in dogs, and also that the blood endotoxin concentration could be used as a marker to determine prognosis. PMID- 9853313 TI - Apoptosis induction of POS canine osteosarcoma cells by vitamin D and retinoids. AB - Vitamin D3: 1-alpha, 25(OH)2D3 (calcitriol), 22-oxa-1,25(OH)2D3 (OCT), cholecalciferol (vitamin D3), and retinoids: all-trans retinoic acid (ATRA) and 9 cis retinoic acid, induced morphological changes in POS canine osteosarcoma cells into elongated, spindle or fibroblast like-shaped cells, and apoptotic like cell death characterized by cell shrinkage, condensation and margination of the nucleus for all drugs at 10(-6)M-10(-9)M after 72 to 120 hr culture. Apoptosis as shown by DNA laddering was induced at 48 hr by all drugs at 10(-6)M, 10(-7)M at 96 hr, 10(-8)M and 10(-9)M at 120 hr respectively. These vitamins are suggested to adjunct antineoplastic agents in canine osteosarcoma therapy by induction of apoptosis. PMID- 9853315 TI - Seroprevalence of infectious bursal disease virus in free-living wild birds in Japan. AB - Serum samples collected from 739 free-living wild birds of 44 species from Gifu, Mie and Hyogo Prefectures in Japan during the period 1989 to 1997 were tested for antibodies to infectious bursal disease virus (IBDV) serotypes 1 and 2 by a virus neutralization test. Serological evidence of infection with serotypes 1 and 2 was found in 15 (2%) of the sera of 6 species and 36 (4.9%) of the sera of 11 species, respectively. Antibodies to IBDV were detected from both sedentary and migratory species. These findings suggest that free-living wild birds have an important role in the natural history of IBDV. These findings raise the possibility that the IBDV prevalent in the breeding grounds of these birds in other countries could be imported by the migratory species. This is the first report of an extensive serological survey of IBDV in wild birds. PMID- 9853314 TI - Seroepidemiological survey of feline retrovirus infections in domestic and leopard cats in northern Vietnam in 1997. AB - The prevalence of infections with three feline retroviruses (feline immunodeficiency virus (FIV), feline leukemia virus (FeLV) and feline syncytial virus (FSV)) was examined in northern Vietnam in 1997. We collected a total of 77 blood samples from 69 domestic and 8 leopard cats, and examined the presence of anti-FIV and FSV antibodies and FeLV p27 antigen in the plasma samples by the indirect immunofluorescence and/or two commercial kits. None of the samples was positive for FIV and FeLV. The overall positive rate of FSV was 31% and the positive rates among the domestic and leopard cats were 29 and 50%, respectively. We isolated FSV from peripheral blood mononuclear cells of 6 domestic and one leopard cats. PMID- 9853316 TI - Production of exfoliative toxin A by Staphylococcus aureus isolated from mastitic cow's milk and farm bulk milk. AB - The production of exfoliative toxins A and B (ETA and ETB) by Staphylococcus aureus isolated from mastitic cow's milk and farm bulk milk was examined by the reverse passive latex agglutination method (RPLA). ETA was detected in 2 (1.2%) of 162 isolates from mastitic cow's milk and in 1 (0.6%) of 166 isolates from farm bulk milk. RPLA titers of these isolates were much lower than in human isolates. No ETB was detected in any of the isolates tested. These ETA-positive isolates belonged to bovine ecovar. They were non-typable using the international phage set for human strains. When these ETA-positive isolates were subcutaneously inoculated into neonatal mice, general exfoliation of the epidermis accompanied by the so-called Nikolsky sign was not recognized. By the immunoblotting and PCR methods, however, ETA and eta gene were recognized in the ETA-positive isolates from mastitic cow's milk and farm bulk milk. These data suggest that ETA is also produced by bovine isolates of S. aureus, but in smaller quantities. PMID- 9853317 TI - Tracing marine biomass into tidal freshwater ecosystems using stable sulfur isotopes. PMID- 9853318 TI - On the reintroduction of the Mhorr gazelle in Tunisia and Morocco. AB - The Mhorr gazelle (Gazella dama mhorr) became extinct in the wild in the 1960s. The current world zoo population amounts to some 240 animals, going back to only 12 founders. In a cooperate international operation Mhorr gazelles of European zoo populations were successfully reintroduced to national parks and nature reserves in Morocco and Tunisia in the years 1990/1992. The genetical and ecological prerequisites as well as the breeding aspects are discussed. PMID- 9853319 TI - Should 'single-use' nebulizers for aerosol lung scans be re-used? PMID- 9853320 TI - Nuclear cardiology in the UK: British Nuclear Cardiology Society survey 1994. AB - This study surveyed practice in nuclear cardiology in the UK in 1994. A questionnaire was sent to 219 centres performing nuclear imaging asking for details of current practice in nuclear cardiology. Replies were received from 192 centres (88%). Activity in performance of myocardial perfusion imaging (MPI) and radionuclide ventriculography (RNV), anticipated changes in activity, differences between regional and district general hospitals, technical imaging parameters and referral sources were surveyed. Of the responding centres, 125 (65%) performed nuclear cardiology. More regional centres (85 vs 55%, P < 0.0003) performed a higher proportion (62 vs 24%, P < 0.001) of nuclear cardiology activity compared with district general hospitals. Nuclear medicine activity was estimated at 9.3 scans/1000/year, of which 8.9% was cardiology (0.82/1000/year; MPI, 0.56/1000/year; RNV, 0.26/1000/year). A comparison with previous surveys showed a significant increase of 24% in nuclear cardiology since 1988, with a strong rise in MPI (350%); however, RNV has fallen by 47%. Myocardial perfusion activity in the UK remains very low (25 and 5% for MPI and RNV respectively) when compared with the average of 2.2/1000/year for Europe and 10.8/1000/year for the USA. In conclusion, MPI has increased on average by 23% per annum (compound rate) since 1988, but in 1994 was still only 32% of the British Cardiac Society target of 2.6 scans/1000/year. Proper resourcing for capital expenditure on new equipment and new staff will be important to maintain momentum in closing the gap. Also important is clinical understanding, as already implemented by including nuclear cardiology in guidelines for specialist cardiology training. PMID- 9853321 TI - Effect of 360 degrees and 180 degrees rotation SPET acquisitions on myocardial polar map: comparison of 201Tl-, 99Tcm- and 123I-labelled radiopharmaceuticals. AB - Both 360 degrees and 180 degrees rotation acquisition methods have been used in myocardial single photon emission tomography (SPET) studies. We compared both methods using 201Tl, 99Tcm and 123I radiopharmaceuticals with phantoms and clinical models. Myocardial phantom studies with anterior and inferior defects were performed using 201Tl, 99Tcm and 123I. Clinical models of 14 typical situations, including normal subjects, patients with anterior or inferior defects and a high right hemi-diaphragm, were studied. The radiopharmaceuticals were 201Tl, 99Tcm-sestamibi, 123I-BMIPP and 123I-MIBG. Four sets of 180 degrees anterior rotation data with starting angles of (A) posterior, (B) LPO 30 degrees, (C) LPO 60 degrees and (D) left lateral direction were generated and compared with 360 degrees rotation SPET. A polar map display was used for quantification. In phantom studies, the defect contrast on the map was higher in the anterior defect with 180 degrees rotation than with 360 degrees rotation. However, it was decreased in the inferior defect, particularly with 201Tl, because of decreased wall activity around the defect. In the patient model with anterior or inferior defects, the defect contrast was improved with 180 degrees SPET by up to 10%. A slight decrease in the normal region was also noted in the 180 degrees reconstruction. The effect of diffuse liver activity on the inferior region depended on the rotation range. A patient with a high right hemi-diaphragm showed a lower inferior count with 360 degrees SPET. In conclusion, the 360 degrees acquisition was superior to the 180 degrees acquisition in the phantom with defects. Clinically, the quantitative differences in radionuclide types (99Tcm, 123I or 201Tl) were not significant for quantifying a moderate degree (50-60% of peak count) of defect. However, we note quantitative variation depending on the rotation range in the 180 degrees method. PMID- 9853322 TI - Basic competencies for the nuclear medicine technologist. British Nuclear Medicine Society Technology Group. PMID- 9853323 TI - Clinical relevance of 201Tl-chloride SPET in the differential diagnosis of brain tumours. AB - Magnetic resonance imaging (MRI) and computed tomography (CT) may not be reliable in the differential diagnosis of tumour necrosis, scar and recurrent tumour. We compared 201Tl-chloride SPET with CT and MRI for the differential diagnosis of these cerebral lesions. Brain SPET was performed in 40 patients after the intravenous injection of 201Tl-chloride. All 40 patients also had a CT or MRI scan, and a histological diagnosis was available for 27 of the patients. For each patient, the ratio of counts in the lesion region of interest (ROI) to counts in the contralateral ROI was calculated and found to be between 0.58 and 9.60. The ratios for high-grade gliomas, metastases and meningiomas were high (> 2.7), especially in tumours with good vascularization. A low ratio (< 1.7) was noted in patients with low-grade astrocytoma, necrosis or ischaemic lesions. There were two exceptional cases of ischaemic lesions in the luxury perfusion stage (ratios of 3.61 and 3.87), as verified by HMPAO-SPET. We found that 201Tl-chloride SPET helps to differentiate between malignant tumours, poorly vascularized benign lesions and necrosis. Differentiation between low-grade astrocytoma and non malignant lesions was not possible, but there was a trend towards differentiating between low-grade astrocytoma and ischaemic infarction. The timing of the investigation is important to avoid false-positive results in hyperperfused ischaemic tissue. PMID- 9853324 TI - Ultrasonographic versus scintigraphic measurement of thyroid volume in patients referred for 131I therapy. AB - The activity of 131I to be administered as therapy to patients with thyroid disease is usually calculated from 24 h radioiodine uptake and thyroid volume. The aim of the present study was to compare thyroid volume, measured by scintigraphy and ultrasonography, to evaluate the impact of these methods on the calculated 131I dose. Forty patients (20 with diffuse toxic goiter and 20 with multinodular toxic or nontoxic goiter) were investigated. On the same day, thyroid volume was measured by ultrasonography (using transverse scans at 5 mm intervals) and by scintigraphy, using either the ellipsoid formula (SC-E: [symbol: see text]/6 x height x width x depth) or the Himanka formula (SC-H: 0.33 x (planimetric surface in pixels)3/2). With ultrasonography, the size of diffuse goiters was smaller than that of nodular goiters (median values and range: 18 ml (11-46) and 50 ml (14-198) respectively, P < 0.001). Both scintigraphic methods, however, failed to demonstrate a significant difference between diffuse and nodular goiter size. In patients with diffuse goiter, thyroid volume measured by SC-E did not differ from that measured by ultrasonography, whereas thyroid size was overestimated by 53% using the Himanka formula. In contrast, in patients with nodular goiter, thyroid volume measured by SC-H did not differ from that measured by ultrasonography, whereas the ellipsoid formula underestimated thyroid size by 48%. The overestimation of diffuse goiter size by the Himanka formula resulted in a relatively modest median excess of 96 MBq (range -118 to +248 MBq) of the calculated 131I dose. The underestimation of nodular goiter size by the ellipsoid formula resulted in a calculated dose that was 278 MBq lower (range -1624 to +141 MBq). The median calculated 131I dose based on the Himanka formula was not different from that based on ultrasound, but large differences in calculated 131I dose (up to 1280 MBq) were found in individual cases. In conclusion, thyroid volume can be assessed with accuracy by scintigraphy using the ellipsoid formula in patients with diffuse goiter. Wide differences, however, are observed in the size of nodular goiters measured by scintigraphy and ultrasonography. PMID- 9853325 TI - Relationship between extraosseous accumulation in bone scintigraphy with 99Tcm HMDP and histopathology. AB - We assessed the relationship between 99Tcm-HMDP extraosseous accumulation (EOA), the histopathology of primary lesions and various laboratory findings. In 155 of 4824 patients, 163 EOA were noted. Of these, 33.7% were in the abdomen, 27.6% in the chest, 22.7% in the extremities, 9.8% in the pelvic area and 6.1% in the head and neck area. We found that 72.4% of EOA were due to malignant processes and 27.6% to benign processes. In the abdomen, 36.6% of the EOA were due to hepatocellular carcinoma and intestinal carcinoma. In the chest, 46.7% of EOA were a result of breast carcinoma. In the extremities, 28.7% of the EOA were due to sarcoma. The mean white blood cell count was elevated (8.0 +/- 6.3 x 10(3)) in patients with malignant processes. The mean serum haemoglobin and haematocrit in benign and malignant processes, for both males and females, were below normal values. All other laboratory findings were within normal limits. Significant differences in serum haemoglobin and haematocrit were noted between male and female patients with benign processes (P = 0.04 and P = 0.04, respectively). No other significant differences between benign and malignant processes were noted. Therefore, EOA is more frequently associated with malignant processes of primary lesions and is often accompanied by leukocytosis and anaemia. PMID- 9853326 TI - Out-of-hours weekend scintigraphy: luxury or necessity? PMID- 9853328 TI - The role of nuclear medicine in the post-infarction period. PMID- 9853329 TI - Detection of lung lesions and lymph nodes with 201Tl SPET. AB - The aim of this study was to characterize lung lesions by means of the uptake, retention and washout of 201Tl, and to quantify the sensitivity of 201Tl SPET (single photon emission tomography) in detecting mediastinal and hilar lymph nodes. A total of 68 patients with lung lesions (tumours and inflammation) underwent both helical computed tomography (CT) and 201Tl SPET examinations. Uptake, retention and washout parameters were calculated from early and delayed 201Tl SPET by drawing regions of interest over the primary lesion and the background of the lung. The findings on 201Tl SPET were compared with those on CT and surgical findings. A significant difference in the retention index (RI) was observed between primary malignant and benign lesions. In detecting lymph nodes, the sensitivity, specificity and negative predictive value were 83%, 60% and 90% respectively for early 201Tl SPET, and 50%, 80% and 80% respectively for delayed 201Tl SPET. 201Tl SPET with semi-quantitative analysis, especially RI, helps to differentiate between malignant and benign lung lesions. Lymph nodes larger than 1 cm on CT but not showing 201Tl uptake could be considered non-metastatic lesions. PMID- 9853330 TI - Follow-up of patients with superior vena cava syndrome by functional analysis of radionuclide venography. AB - In addition to imaging, radionuclide venography can be used for studying haemodynamic changes in superior vena cava syndrome (SVCS) by using the indices of transit time (TT), time of half peak count (TH) and peak count ratio (PC ratio). The objective of this study was to examine the utility of applying these indices, alongside images, in the post-therapy assessment of SVCS patients. Radionuclide venography was performed twice on 36 patients with SVCS due to thoracic malignancies, 29 of whom received specific anti-cancer therapy between the two studies (Group I); the other 7 did not receive such therapy (Group II). On the images, 12 patients in Group I showed improvement, one a deterioration and 16 no change; in Group II, 4 showed a deterioration and 3 no change. Using the indices, nearly all patients in Group I had decreased post-therapy values indicating haemodynamic improvement. A significant difference was seen between the mean (+/- S.E.) pre- and post-therapy values for TT (pre vs post: 6.2 +/- 0.7 vs 2.7 +/- 0.3 s) and TH (pre vs post: 16.9 +/- 2.3 vs 7.9 +/- 0.8 s) (P < 0.001). Although the PC ratio showed a non-significant decrement, it related to the status of collateral change. The above indices provide clinically valuable information about the haemodynamic status of patients with SVCS and can be used for their post-therapy evaluation. PMID- 9853331 TI - Rhenium-188 microspheres: a new radiation synovectomy agent. AB - Radiation synovectomy is efficacious in controlling the symptoms of rheumatoid arthritis. However, the procedure is not widely used because of concerns about leakage of radiopharmaceuticals from the treated joints. Leakage can be minimized by selecting particles of an appropriate size. In this study, we labelled microspheres with 188Re and analysed its biodistribution after intra-articular injection in rabbits with antigen-induced arthritis. Gamma camera imaging was performed to quantify the mean retention of 188Re in the knees. The mean retention of 188Re was 98.7, 94.6 and 93.6% at 1, 24 and 48 h, respectively. The biodistribution data revealed very low radioactivity in all organs at different times, which suggests the leakage of radiotracer from the knee was negligible. Our preliminary results indicate that 188Re microspheres are a potentially effective radiopharmaceutical for radiation synovectomy. PMID- 9853332 TI - Simultaneous biplane first-pass radionuclide ventriculography using 99Tcm tetrofosmin: a comparison with magnetic resonance imaging. AB - First-pass radionuclide ventriculography (FPRNV) using 99Tcm-labelled myocardial perfusion agents allows the assessment of myocardial function and perfusion simultaneously. We have assessed the feasibility of biplane FPRNV using 99Tcm tetrofosmin, and have validated global and regional functional measurements by comparison with magnetic resonance imaging (MRI). FPRNV was performed at rest in 18 patients referred for assessment of known or suspected coronary artery disease (5 with previous myocardial infarction). A dual-headed camera was used to acquire RAO and LAO projections simultaneously. Left ventricular ejection fraction (LVEF) was calculated using standard methods and regional wall motion was evaluated visually in five myocardial segments using a 4-point scale and also by Fourier analysis. Cine MRI was performed in four oblique planes, LVEF was calculated using a biplane area-length method and regional motion was assessed visually in a similar fashion to FPRNV. Agreement between the techniques for LVEF was good using RAO FPRNV (mean +/- S.D. difference = 0.7 +/- 4.7%), but less good in the LAO projection (difference = 10.5 +/- 7.1%). Wall motion was normal by both FPRNV and MRI in 5 patients without CAD and 3 of the 13 patients with CAD. In the remaining 10 patients, wall motion by MRI was abnormal in 20 segments; FPRNV with visual analysis was abnormal in 8 patients (80%) and 16 (80%) segments. Fourier analysis showed regional abnormalities in 7 patients (70%) and 13 (65%) segments. There was good agreement (kappa = 0.68) between FPRNV (visual analysis) and MRI for wall motion scores, but moderate agreement (kappa = 0.55) between Fourier analysis and MRI. Thus, LVEF measured by FPRNV in the RAO projection agrees well with MRI. Normal regional wall motion is accurately identified, but regional abnormalities are better assessed with MRI. PMID- 9853333 TI - The value and correlates of left ventricular cavity assessment in dipyridamole 201Tl SPET studies. AB - Left ventricular cavity (LVC) enlargement during SPET dipyridamole 201Tl myocardial perfusion imaging studies is a proven marker of severity of coronary artery disease. Nevertheless, the influence of the extent of myocardial infarct and ischaemia on the degree of LVC enlargement both at rest and with dipyridamole has not been clearly analysed. One hundred and one patients were studied by both dipyridamole myocardial perfusion imaging and radionuclide ventriculography within 1 week. The left ventricular ejection fraction (LVEF) was 57 +/- 9 in normal resting LVC patients (group I), 43 +/- 8 in mild LVC enlargement patients (group II) and 28 +/- 5 in moderate-to-severe LVC enlargement patients (group III). The number of fixed defects was increased in patients in group II and group III, but there was no significant differences in the number of ischaemic segments among groups. The number of ischaemic segments was much higher in patients with transient cavity dilatation than those without cavity change; nonetheless, both LVEF and the numbers of fixed segments were unchanged. The degree of LVC enlargement at rest strongly reflects the resting left ventricular systolic function as well as the extent of previous myocardial infarct. On the other hand, transient cavity dilatation during dipyridamole infusion can only reflect the extent of viable myocardium at risk. PMID- 9853334 TI - Cardiac sympathetic nervous disintegrity is related to exercise intolerance in patients with chronic heart failure. AB - The aim of this study was to clarify the relationship between cardiac sympathetic nervous activity assessed by 123I-MIBG imaging and exercise capacity in patients with congestive heart failure. The subjects were 24 patients with heart failure (NYHA class II to III) and 7 normal controls. A dose of 148 MBq of 123I-MIBG was administered intravenously, and 5-min anterior planar images were obtained 20 min (early) and 4 h (delayed) after the injection of 123I-MIBG. Regions of interest were placed over the left ventricle and mediastinum, and the heart-to-mediastinum ratio (H/M ratio) was calculated as a fraction of the mean counts per pixel in the heart divided by those in the mediastinum. The washout rate from the myocardium was determined by: (early counts delayed counts) x 100/early counts (%). Treadmill exercise tests were performed using a ramp method within 1 week of the 123I-MIBG studies. During exercise, expired gas was analysed, and peak VO2 and VO2 at the anaerobic threshold (ATVO2) were measured. A significant linear correlation was found between the H/M ratio on delayed 123I-MIBG images and exercise duration (r = 0.48, P < 0.05), peak VO2 (r = 0.49, P < 0.05) and ATVO2 (r = 0.56, P < 0.01). The washout rate of 123I-MIBG was inversely correlated with exercise duration (r = -0.45, P < 0.05), peak VO2 (r = -0.51, P < 0.01), and ATVO2 (r = -0.50, P < 0.05). In conclusion, enhanced cardiac sympathetic nervous activity relates to exercise intolerance in patients with congestive heart failure. The non-invasive assessment of cardiac sympathetic nervous activity by 123I-MIBG can predict exercise capacity in human heart failure. PMID- 9853335 TI - Physical performance of the Siemens E.CAM gamma camera. AB - We evaluated the physical performance of the new Siemens E.CAM gamma camera. The camera is a dual-headed, large detector system with an open gantry dedicated to planar and to multislice single photon emission tomographic (SPET) imaging of the whole body. The energy-independent digital detectors with high-quality photo multiplier tubes seem to keep the physical performance of the E.CAM camera very stable. The imaging resolution is very good. The SPET resolution is 9.1 mm full width at half maximum in scatter for high-resolution collimators with a rotational radius of 15 cm. Both planar (< or = 2%) and SPET uniformity (< or = 4%) are excellent. The detectors can be configured at variable angles which allows flexibility, especially in heart studies. Autocontouring performs whole body and SPET scans as close to the patient as possible. The patient couch, with its very low gamma-ray attenuation, is firm and can be lowered to make patient loading easier. Camera tuning, calibration, acquisition and processing are controlled by two Power Macintosh 8500 computers. The ICON software is easy to use (a mouse-driven system). However, acquisition interrupts and autocontouring errors were noted during our initial experience. The image quality of routine bone and heart scans was excellent. The E.CAM system allows fast and accurate planar and SPET imaging both in clinical practice and research. PMID- 9853336 TI - 99Tcm-labelled chimeric human/mouse anti-granulocyte antibody bone marrow scintigraphy: a preliminary clinical study. AB - Bone marrow scintigraphy using 99Tcm-labelled chimeric anti-granulocyte antibody (anti-NCA-95) was performed in 17 patients with haematological disorders and skeletal metastases. Chimeric anti-NCA-95 antibody (chNCA95 Ab, 0.2 mg) labelled with 444 MBq 99Tcm was administered to obtain bone marrow images 4 h post injection. One week later, an 111In-chloride bone marrow scan was performed on nine patients with haematological disorders. Lumbar bone marrow-to-background (L/B) and ilium-to-background (I/B) uptake ratios were calculated for each scan. In six patients with suspected skeletal metastases, 99Tcm-HMDP bone scans were performed. No patient had any adverse reaction or any immune reaction over 20 weeks. In the patients with haematological disorders, the L/B and I/B ratios of the 99Tcm-chNCA95 Ab scan were 3.41 +/- 0.90 and 1.23 +/- 0.31, whereas those of the 111In-chloride scan were 1.58 +/- 0.32 and 1.00 +/- 0.32, respectively. In assessing findings of irregular central bone marrow uptake and peripheral expansion of the bone marrow, the 99Tcm-chNCA95 Ab scan was much better than the 111In-chloride scan. In the six patients with suspected skeletal bone metastases, three true-positive and two true-negative results were observed. This preliminary study has revealed that 99Tcm-chNCA95 Ab scanning is safe and useful in the diagnosis of haematological disorders and skeletal metastases. PMID- 9853337 TI - Preparation of 99Tcm-MAG3: no confirmation that sodium chloride injections from plastic containers affect radiochemical purity. AB - Reports have suggested that when sodium chloride injections from a plastic ampoule are used during the preparation of 99Tcm-mercaptoacetyltriglycine (99Tcm MAG3), the radiochemical purity of the final product might be reduced. A study was therefore undertaken to examine the effect of sodium chloride injections from five manufacturers on the radiochemical purity and stability of 99Tcm-MAG3. One sodium chloride injection was supplied in a glass vial, three in plastic ampoules and one in a plastic infusion bag. Three batches of sodium chloride injections from each manufacturer were tested. The radiopharmaceutical was prepared at a radioactive concentration of 1.1 GBq in 10 ml according to the instructions of the manufacturer of TechneScan MAG3. Analysis of radiochemical purity was performed by high-performance liquid chromatography immediately after preparation and 6 h later. Using 95% as the minimum acceptable radiochemical purity, all the products were satisfactory over the 6 h test period. No manufacturer's sodium chloride injection was found to have a statistically significant effect on the radiochemical purity. Based on the 15 batches of sodium chloride injection tested, this study cannot confirm that sodium chloride injections from a plastic container affect the radiochemical purity of 99Tcm-MAG3. However, in view of the known sensitivity of some 99Tcm radiopharmaceuticals to external influences, it is probably good practice to test radiochemical purity when new batches of ancillary materials, such as sodium chloride injections, are introduced. PMID- 9853338 TI - Stability of Adenoscan (adenosine 3 mg ml-1) in plastic syringes. AB - Adenosine is an effective and safe agent for myocardial stressing in nuclear medicine. It is commercially available as Adenoscan solution (adenosine 3 mg ml 1) and can be conveniently administered to patients via an anaesthesia syringe pump. The solution is dispensed in plastic syringes prior to administration to the patient. The aim of this study was to assess the stability of Adenoscan in plastic syringes to determine if it is possible to pre-dispense the solution and to store dispensed syringes ready for administration to patients. A 4 week stability analysis using high-performance liquid chromatography was carried out on Adenoscan that had been pre-dispensed into polypropylene syringes and stored at 2-8 degrees C. It was found that the concentration of adenosine in the pre dispensed syringes remained stable during the 4 week period. PMID- 9853339 TI - Comparison of HPLC, TLC and minicolumn for measuring the radiochemical purity of 99Tcm-Q12. AB - TechneScan Q12 (99Tcm-Q12) is a new agent for clinical myocardial perfusion imaging. A product with high radiochemical purity is essential for high-quality imaging. We compared three methods of radiochemical purity analysis for 99Tcm Q12: thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and minicolumn. Thin-layer chromatography resulted in lower purity (87 +/- 8%) than the minicolumn method (aluminium oxide minicolumn, ethanol) (95 +/- 4%). The HPLC method resulted in the lowest purity (79 +/- 11%). The main impurity was a polar compound, but three further impurities were found using HPLC. Using HPLC, we found the percentage of the parent compound (99Tcm-Q12) in plasma at 40-90 min post-injection to be 19.3 +/- 6.2%. We suggest that gradient HPLC is the most effective method for the analysis of the radiochemical purity of 99Tcm-Q12, and that it can be used to determine the concentration of 99Tcm-Q12 in plasma. PMID- 9853340 TI - Radiopharmaceutical kit modification. PMID- 9853341 TI - The influence of generator eluate on the radiochemical purity of 99Tcm-sestamibi prepared using fractionated Cardiolite kits. AB - The reconstitution of fractionated Cardiolite kits with sodium 99Tcm pertechnetate eluted from a wet-column 99Mo/99Tcm generator results in a preparation that contains elevated amounts of a polar radiochemical impurity, which is normally present in only trace amounts when such kits are reconstituted with eluate from a dry-column generator. A consequence of this impurity, which appears to be an organotechnetium compound, is that it causes a positive bias in several quality control methods for 99Tcm-sestamibi, including that recommended by the manufacturer. This impurity was isolated, and its biodistribution determined in Wistar rats. Two alternative methods for the preparation of 99Tcm sestamibi from fractionated Cardiolite kits using eluate from a wet-column generator were investigated. A rapid procedure, based on a C18 solid phase extraction (SPE) cartridge, for measuring the radiochemical purity of 99Tcm sestamibi samples, which had been prepared using fractionated Cardiolite kits, was developed and compared with two other methods including high-performance liquid chromatography (HPLC). Over a radiochemical purity range of 49-98%, the values obtained using our quality control method deviated from those obtained using HPLC by 1.1 +/- 1.3% (n = 26). In contrast to some of the rapid quality control methods assessed in our investigation, the SPE technique was not biased by the presence of the organotechnetium impurity. PMID- 9853342 TI - Regional glucose utilization in infarcted and remote myocardium: its relation to coronary anatomy and perfusion. AB - We studied the relationship between coronary anatomy, perfusion and metabolism in myocardial segments exhibiting transient and persistent perfusion defects on stress/rest 99Tcm-MIBI single photon emission tomography in 35 patients (31 males, 4 females, mean age 56 +/- 7 years) with a previous myocardial infarction. Quantitative coronary angiography and assessment of myocardial perfusion reserve and glucose metabolism were performed within 1 week of one another. Perfusion was assessed by SPET after the intravenous injection of 740 MBq of 99Tcm-MIBI at rest and after exercise. Regional myocardial glucose metabolism was assessed by position emission tomography at rest (200 MBq of 18F-2-deoxyglucose, FDG) after an overnight fast with no glucose loading. All 35 patients exhibited persistent perfusion defects consistent with the clinically identified infarct site, and 27 (77%) also showed various degrees of within-infarct FDG uptake; 11 patients developed exercise-induced transient perfusion defects within, or in the vicinity of, 15 infarct segments and resting FDG uptake was present in 10 of these segments (67%). Five patients also showed exercise-induced transient perfusion defects in nine segments remote from the site of infarct: resting FDG uptake was present in six of these regions (67%). Finally, nine patients had increased glucose uptake in non-infarcted regions not showing transient perfusion defects upon exercise testing and perfused by coronary arteries with only minor irregularities. Our results confirm the presence of viable tissue in a large proportion of infarct sites. Moreover, FDG uptake can be seen in regions perfused by coronary arteries showing minor irregularities, not necessarily resulting in detectable transient perfusion defects on a MIBI stress scan. Since the clinical significance of such findings is not clear, further studies should be conducted to assess the long-term evolution of perfusion, function and metabolism in non revascularized patients of those remote areas which are apparently normally perfused, but show abnormal fasting FDG uptake after myocardial infarction. Such studies may have important implications for the management of post-infarct patients, as the preservation of coronary vasodilator reserve and myocardial metabolism in remote myocardium may be seen as an additional goal in the treatment of such patients. PMID- 9853343 TI - Stress and rest myocardial SPET quantification with 201Tl and 99Tcm-tetrofosmin: a comparison. AB - 99Tcm-tetrofosmin is a new myocardial perfusion agent that has excellent physical and pharmacokinetic characteristics for performing tomographic myocardial perfusion studies. The aim of this study was to compare the behaviour of 99Tcm tetrofosmin and 201Tl in the assessment of ischaemia and viability in patients with previous myocardial infarction. Twenty consecutive patients who had suffered infarction and been referred for assessment of ischaemia and myocardial viability were enrolled into the study. Each patient underwent two stress tests performed no more than 10 days apart, one with a 201Tl exercise-reinjection-redistribution method and one with a 99Tcm-tetrofosmin short protocol (exercise-rest). The results were quantified using polar maps to represent images for stress, rest and reversibility. The post-stress images showed there was a slight tendency to overestimate defect size with 99Tcm-tetrofosmin in the lateral region (P < 0.006). We found no significant differences between the two tracers when comparing reversibility. The same defect size at rest were obtained when the maps for 201Tl with uptake of 50% were compared with those for 99Tcm-tetrofosmin with uptake of 45%. Agreement between the two agents for assessment of viability was 93%. We conclude that the quantitative assessment of myocardial ischaemia and uptake of 99Tcm-tetrofosmin at rest are comparable to those obtained with 201Tl in patients who have suffered myocardial infarction. PMID- 9853344 TI - Metabolic (PET) and receptor (SPET) imaging of well- and less well-differentiated tumours: comparison with the expression of the Ki-67 antigen. AB - [111In-DTPA-D-Phe1]-pentetreotide has been shown to localize well-differentiated and slowly growing neuroendocrine tumours, whereas increased FDG uptake is associated with malignancy. This prospective study explores the role of metabolic (PET) and receptor (SPET) imaging in well- and less well-differentiated tumours- gastroenteropancreatic (GEP) tumours, medullary thyroid carcinomas (MTC) and thymic carcinomas--in comparison with the expression of the Ki-67 antigen. Ten patients with GEP tumours, five with MTC and five with thymic carcinomas were studied. Prior to PET, somatostatin receptor scintigraphy (SRS) was performed in all patients. Sixty minutes after the intravenous administration of 18F-FDG (370 MBq), whole-body PET was performed. In addition, the resected tissues were prepared for immunocytochemistry examination (cell cycle-associated Ki-67 antigen). Preoperative SRS detected multiple primary tumours and metastatic lesions in four patients with well-differentiated carcinoids (low Ki-67 expression). Whole-body PET demonstrated normal distribution of FDG in all of these patients. In patients with recurrent MTC and rapidly increasing CEA levels, SRS showed no in vivo somatostatin receptor expression, whereas whole-body PET localized 24 locoregional lymph node metastases with increased FDG uptake. Immunocytochemistry of the resected lymph nodes demonstrated high Ki-67 expression associated with a high proliferative activity. Similar results in receptor scintigraphic and metabolic behaviour were obtained from patients with metastasizing thymic carcinomas (high Ki-67 expression). In conclusion, SRS has been shown to localize well-differentiated GEP tumours. In contrast, FDG PET is only valuable for predicting malignancy in less well-differentiated GEP tumours and malignant MTC associated with rapidly increasing CEA levels. Therefore, an additional 18F-FDG PET procedure should only be performed if SRS is negative. Furthermore, our preliminary results suggest that increased FDG metabolism reflects the invasiveness of thymic carcinomas. PMID- 9853346 TI - Detection of lung and chest tumours using 99Tcm-tetrofosmin: comparison with 201Tl. AB - The aim of this study was to assess the usefulness of 99Tcm-tetrofosmin for detecting tumours of the lungs and adjacent structures and to compare the results with those for 201Tl. In the 18 patients studied, there were 30 lesions in total. Dual-isotope SPET acquisitions were performed 10 min and 3 h after the administration of 99Tcm-tetrofosmin and 201Tl. Image quality and semi quantitative parameters were evaluated for both tracers. All lesions were detected by the two radiopharmaceuticals on both the early and delayed images. In seven patients, the image quality of 99Tcm-tetrofosmin was better than that of 201Tl. For the detection of metastatic lymph nodes, the delayed 201Tl and the early 99Tcm-tetrofosmin images were superior. No significant difference was observed between the uptake ratios of the two tracers. 99Tcm-tetrofosmin showed significantly (P < 0.001) higher washout than 201Tl from both the lesions and normal lungs, although washout of 201Tl from normal lungs was significantly higher than that from the lesions. In conclusion, 99Tcm-tetrofosmin may be helpful for detecting tumours of the lungs and adjacent structures, but has limited applicability in differentiating between malignant and benign tumours and may not be used instead of 201Tl to detect malignant tumours of the lungs. PMID- 9853345 TI - Use of 99Tcm-MIBI in the assessment of patients with suspected recurrent breast cancer. AB - A prospective trial was performed to assess the accuracy of 99Tcm-methoxyisobutyl isonitrile (99Tcm-MIBI) scintimammography and X-ray mammography in 18 patients (mean age 58 years, range 46-79 years) with suspected recurrent breast cancer in the breast and/or loco-regional tissues. All patients had been diagnosed to have breast cancer 1-23 years before scintimammography. Two patients had undergone mastectomy, so that a total of 34 breasts were studied with X-ray mammography and with prone-lateral and anterior scintimammography. Any abnormalities on X-ray mammograms and scintimammograms were noted, as was any additional local or nodal uptake of 99Tcm-MIBI. The nature of any lesion seen with either modality, or in which there was a clinical suspicion of recurrence, was confirmed by cytological or histological examination of tissue samples. There were nine breasts with recurrent cancer in eight patients. X-ray mammography identified six of these cancers. 99Tcm-MIBI scintimammography identified eight of nine recurrent breast cancers (the one cancer not seen was positive on X-ray mammography). In the 26 breasts without cancer, two were reported as having changes consistent with cancer and one was reported equivocal. There were three false-positive and one equivocal scintimammograms. The accuracy of 99Tcm-MIBI scintimammography within the breast (85%) was similar to X-ray mammography (82%). Axillary lymph node recurrence occurred in five patients, four of whom were positive with 99Tcm-MIBI. An additional patient had focal uptake of 99Tcm-MIBI at the site of recurrent cancer on the anterior chest wall. In conclusion, 99Tcm-MIBI scintimammography may provide useful complementary information in patients with suspected recurrence of cancer in the breast or loco-regional tissues. PMID- 9853347 TI - Evaluation of 99Tcm-HIG as a lymphoscintigraphic agent in rabbits. AB - In this pilot study, 99Tcm-labelled human immunoglobulin G (99Tcm-HIG) was evaluated as a lymphoscintigraphic agent in five rabbits. It was injected intradermally into the web space of the hind legs of the rabbits (37 MBq/0.1 ml). Sequential scintigrams were obtained using a gamma camera for 120 min. The injection site and the hind legs were massaged post-injection. Blood samples were obtained at 5, 15, 30, 60, 90 and 120 min. Two of the rabbits were killed after 2 h. Their organs were weighed and tissue specimens were obtained, weighed and counted against a standard using a gamma counter. The lymph channels and the lymph nodes were well visualized on the scintigrams. The background activity was very low, making interpretation easier. About 30% of the injected dose migrated from the injection site by 2 h. The mean popliteal lymph node uptake was 5.71 +/- 4.62% per gram of tissue. The lymph node to other tissue concentration ratios were very high, ranging from 63:1 for the kidneys to 1099:1 for the heart. We conclude that 99Tcm-HIG is a promising new agent for the visualization of the lymphatic system due to its easy labelling procedure, the stability of the label, its widespread availability and good image quality. It may potentially be useful in detecting and evaluating inflammatory lymph nodes. PMID- 9853349 TI - Determination of small objects from pinhole scintigrams. AB - This study assessed the possibility of measuring the linear dimensions of small structures using pinhole scintigraphy. A number of glass objects were made with a spherical, cylindrical or conical shape. Their maximum dimensions (diameters and heights) were 3.5-22.5 mm. These glass objects were filled with 131I, placed inside a plastic neck phantom and imaged using a gamma camera equipped with a pinhole collimator. The source-to-collimator distance was varied from 2 to 12 cm. An algorithm for image segmentation (threshold selection) was used to divide the image into object and background. On the segmented image, the number of non-zero pixels in the direction of the principal axes was multiplied by the appropriate calibration factor to obtain the linear dimensions of the object. Spatial resolution of the pinhole collimator, expressed as the full-width at half-maximum (FWHM), varied from 8 to 10 mm for the range of source-to-collimator distances examined. We found that, for dimensions up to 1.5 x FWHM, finite spatial resolution affects the accuracy of measurement. Non-linear correlation between true and calculated dimensions was used to take the latter into account. Our results are now being used to improve quantitation of remnant thyroid tissue masses for the calculation of radioiodine ablation doses. PMID- 9853348 TI - Baumkuchen map analysis to detect postural cerebral hypoperfusion with upright 99Tcm-HMPAO brain SPET. AB - We have evaluated a polar cortical flow Baumkuchen map (BK map), constructed from projection data by inwardly layering cortical flow stripes from the base to the apex, to detect postural cerebral hypoperfusion using 99Tcm-HMPAO brain SPET. A circular cortical perfusion outline was constructed from sinogram data at a 20% threshold; a ring was then formed by constructing an inner line with a radius equal to half that of the outer line. This ring on the transaxial image was superimposed onto the cortical image, then divided into 60 portions at 6 degrees intervals from the centre. The maximal number of counts per pixel within each sector was determined, translated into polar coordinate profiles and displayed as a polar map. We studied 11 patients with unilateral stenosis of the internal carotid artery or middle cerebral artery (MCA) and 7 patients without stenosis, using upright and supine (rest) 99Tcm-HMPAO brain SPET Cerebral perfusion was visibly altered on the BK maps of all 11 patients with arterial stenosis and on the transaxial images of 8 of them. A semi-quantitative analysis of the BK maps using a ratio of the counts in the regions of interest over the right and left MCA areas, showed that the mean of the ratios was significantly lower in the upright images than in the upright + rest and rest images (P = 0.0044 and 0.0033, respectively). Although the difference in the means of the ratios between the upright and upright + rest images was significant for both the BK map and transaxial image data (P = 0.0044 and 0.0051, respectively), the BK map was accurate in detecting changes in perfusion in the upright position. We conclude that BK map analysis is useful for detecting changes in patients examined using upright brain SPET. PMID- 9853350 TI - Does renal damage occur after the administration of 32P for palliation of pain from skeletal metastases? AB - In normal adults, the maintenance of phosphate balance involves the reabsorption of 85-90% of filtered phosphate by the proximal tubule. At a glomerular filtration rate (GFR) of 125 ml min-1 and a plasma phosphate concentration of 1.3 mmol l-1, the filtered phosphate is approximately 235 mmol day-1. Following intravenous administration, 25-50% of 32P is excreted over 4-6 days in normal subjects. In spite of such extensive renal handling of phosphate and, therefore, of 32P, there are no data in the literature concerning possible 32P-related nephrotoxicity. Adult dosimetry values for the kidney after 32P are reported as 4.8 rad mCi-1 h-1 (0.048 mSev 37 MBq-1 h-1). The entry criteria for 32P therapy insist on a normal serum creatinine value, reflecting awareness of potential renal damage. To answer the fundamental question of whether there is demonstrable renal damage after 32P, we undertook serial measurements of GFR in patients given 32P for treatment of pain from skeletal metastases. Twenty-one patients who had normal pre-treatment renal function as shown by normal serum creatinine values were administered 32P orally in doses ranging from 277.5 to 466.2 MBq, with a mean of 425.5 MBq. Pre-treatment, GFR was estimated with 99Tcm-diethylenetriamine pentaacetate renography using the Gates protocol. Post-treatment, GFR was estimated serially as far as possible, at weeks, 1, 2, 3 and 4 and then every 4 weeks for another 3 months, at which point follow-up ceased. Serum creatinine was assessed pre-treatment and every 2 weeks until the end of follow-up, in addition to all other parameters and a clinical evaluation. Mean pre-treatment GFR was 87.5 ml min-1, with a range of 48.7-110 ml min-1. Not all patients could fulfil the entire follow-up schedule as designed, but we obtained a minimum of four follow-up tests, two before and two after 4 weeks post-treatment. GFR fell to 72% of the pre-therapy value during the first 4 weeks following therapy. By the sixteenth week, however, the mean value had returned to or exceeded the pre treatment value. There was no change in serum creatinine values. At a time when multiple therapies are being considered, this early depression of GFR may be of importance. A closer assessment of altered renal function may be warranted and other sensitive tests of renal damage like microalbuminuria could be used. PMID- 9853352 TI - The importance of 99Tcm-DMSA renal scintigraphy in the follow-up of acute pyelonephritis in children: comparison with urographic data. AB - At present, 99Tcm-dimercaptosuccinic acid (DMSA) renal scintigraphy is the most sensitive examination for the detection of parenchymal damage during acute pyelonephritis (APN) in children. This prospective study had three aims: (1) to evaluate the medium-term evolution of the scintigraphic abnormalities, to find a prognostic criterion of scintigraphic evolution; (2) to assess the correlation between the severity of early or late scintigraphic damage and selected clinical factors; and (3) to compare the permanent scintigraphic renal scars with intravenous urography (IVU) 2 years after the acute infection. Seventy-four children (mean age 32 months), presenting with a first clinical episode of pyelonephritis and an initial scintigraphic abnormality, were included in the study. Patients with a history of urinary tract infection (UTI), uropathy other than vesico-ureteral reflux (VUR) and a relapse of acute pyelonephritis were excluded. All children underwent control scintigraphy (mean 9 months after APN) and 43 had an IVU (mean 26 months after APN). Fifty-seven children (77%) still have scintigraphic abnormalities of varying severity (7 atrophic kidneys). Initial relative DMSA uptake of less than 45% results in a worse scintigraphic prognosis. The age of the child has no bearing on the severity of the initial renal involvement or on the evolution of the scintigraphic abnormalities. The rapid introduction of antibiotics (< 12 h) significantly improves the scintigraphic prognosis (P < 0.01). The presence of reflux (n = 39) leads to more serious initial damage, but we did not find any effect on later evolution in this study, in which all reflux was low grade in nature. Among the 43 children who had an IVU, 5 showed typical urographic and scintigraphic renal scars in the corresponding region and 38 showed a normal IVU with 28 cases of scintigraphic abnormalities. A DMSA scan is more sensitive than IVU for the detection of renal scarring after a first episode of APN. PMID- 9853351 TI - Administered activity of 99Tcm-DMSA for kidney scintigraphy in children. AB - Kidney scintigraphy using 99Tcm-dimercaptosuccinic acid (DMSA) is a common gamma camera investigation in paediatric patients. The present study deals with methods of calculating the activity to be administered to children. Our aim is to find a method of calculation which gives the same count density in the gamma camera images for all ages. Four different methods, based on body weight, height, body surface area and a theoretical model, are compared. The count density in the images of the kidneys was studied retrospectively in 85 patients aged between 3 weeks and 19.5 years. Fourteen young adults aged 22.0-31.9 years were also studied. Two of the calculation methods, based on body surface area and the theoretical model, showed no apparent age dependency (i.e. a constant count density was observed for all ages). The variation between individuals was similar with both methods, but only the body surface area method gave a count density comparable to that for the young adults. As the method based on body surface area is also easy to implement, this method is recommended. PMID- 9853353 TI - Quality assurance in renography: a review. AB - Despite the fact that renography is practised by most nuclear medicine departments, some doubt remains about its clinical utility in certain settings. Unfortunately, lack of standardization has made it difficult to judge whether some reports of poor diagnostic accuracy are due to a fundamental limitation of the test, or simply to sub-optimal implementation. Some important work may therefore have been overlooked. Technical factors, to do with data acquisition and processing, are the most obvious reasons for variability of results between centres, but patient selection can also have a profound effect on diagnostic accuracy in situations where disease prevalence is low (e.g. renovascular hypertension). Belatedly, these issues are being addressed by the nuclear medicine community and this review summarizes the efforts that have been made to date. Published guidelines need to be reinforced by an audit process if real improvements in quality and consistency are to be realized. PMID- 9853354 TI - False-negative bone scan and metastatic breast carcinoma. PMID- 9853355 TI - Increasing the cyclodextrin complexation of drugs and drug biovailability through addition of water-soluble polymers. AB - For a variety of reasons, including cost, production capabilities and toxicology, the amounts of cyclodextrin (CD) that can be included in drug formulations is limited and, thus, it is important to develop methods which can be applied to enhance the complexation efficacy. Moreover, bioavailability of drugs in CD containing vehicles can be rather limited resulting in less than optimal drug delivery. Addition of small amounts of a water-soluble polymer to the aqueous complexation media enhances the CD complexation of drugs. Thus, less CD is needed to dissolve a given amount of drug when a polymer is present than when it is not present in the aqueous complexation media. Furthermore, the bioavailability of drugs is enhanced through addition of polymers. In general, the water-soluble polymers improve both pharmaceutical and biological properties of drug-CD complexes. This polymer enhancement of drug complexation and bioavailability is independent of the physiochemical properties of the drug. Thermodynamic studies and other physicochemical observations indicate that the polymers participate directly in the drug-CD complexation. PMID- 9853356 TI - A novel synthesis of fused pyrazole systems as antimicrobial agents. AB - The pyrazolo[3,4-b]pyridine derivatives 3 could be prepared by condensing compounds 1 with the 3-aminopyrazolone derivative 2. The pyrazolo[5,2-b]-1,3 oxazine derivative 11 and polyfunctionally substituted 1,4-dihydropyridines 15, 18 were also synthesized. Some of the obtained compounds were tested for their antimicrobial activity. PMID- 9853357 TI - Synthesis and biological evaluation of 1,2,5-oxadiazole N-oxide derivatives as hypoxia-selective cytotoxins. AB - Synthesis and biological evaluation of new 1,2,5-oxadiazole N-oxide derivatives with potential cytotoxic effects are described. From the series of compounds tested, compounds 2 and 6 proved to be very active, although non-selective. PMID- 9853358 TI - Investigations of the cross reactivity of isothiazol-3(2H)-one 1,1-dioxides. AB - Mono- and bicyclic isothiazol-3(2 H)-one 1,1-dioxides and tetrahydrosaccharines 7 were produced by oxidation of the 3-unsubstituted isothiazoles 6. Determination of cross reactivity indicates high values for Compounds 7e and 7l. PMID- 9853359 TI - [Basic substituted thieno(3,4-d)pyrimidine carboxylic acids: synthesis via ANRORC reaction starting from 7-cyanthieno(3,2-d)oxazines]. AB - The synthesis of 7-cyan-thieno[3,2-d]-1,3-oxazine 7 is reported. Reaction of 7 with primary amines gives the thieno[3,4-d]pyrimidincarboxylic acids 14 in a unusual ring transformation. Several synthesized compounds were tested as antiallergic agents. PMID- 9853360 TI - Effects of benzohydroxamic acids on L1210 lymphoid leukemia cell nucleic acid metabolism. PMID- 9853361 TI - Penetration of zinc ions from zinc ointment through the skin of suckers (pig) in vitro. PMID- 9853362 TI - Inhibition of neutral metalloendopeptidase and angiotensin-converting enzyme by selected naturally occurring chromone derivatives. PMID- 9853363 TI - [Leiocarposid--lead structure for the quality assurance of Solidaginis virgaureae herba]. PMID- 9853364 TI - Linoleic acid peroxidation--the dominant lipid peroxidation process in low density lipoprotein--and its relationship to chronic diseases. AB - Modern separation and identification methods enable detailed insight in lipid peroxidation (LPO) processes. The following deductions can be made: (1) Cell injury activates enzymes: lipoxygenases generate lipid hydroperoxides (LOOHs), proteases liberate Fe ions--these two processes are prerequisites to produce radicals. (2) Radicals attack any activated CH2-group of polyunsaturated fatty acids (PUFAs) with about a similar probability. Since linoleic acid (LA) is the most abundant PUFA in mammals, its LPO products dominate. (3) LOOHs are easily reduced in biological surroundings to corresponding hydroxy acids (LOHs). LOHs derived from LA, hydroxyoctadecadienoic acids (HODEs), surmount other markers of LPO. HODEs are of high physiological relevance. (4) In some diseases characterized by inflammation or cell injury HODEs are present in low density lipoproteins (LDL) at 10-100 higher concentration, compared to LDL from healthy individuals. PMID- 9853365 TI - Cutinase binding and activity at the triolein-water interface monitored by oil drop tensiometry. AB - Changes of the oil-water interfacial tension resulting from binding of Fusarium solani pisi cutinase and subsequent lipid hydrolysis were investigated using the oil drop technique. An ELISA was developed to determine the amount of cutinase bound to the triolein-water interface after biotinylation of the enzyme. Cutinase irreversibly adsorbs to a maximum value of about 2 mg/m2. A minimal specific activity of 110 mumol/min/mg was calculated for cutinase acting on a single oil droplet, which is close to the activity found for triglyceride emulsions. At a maximum surface load cutinase could generate one monolayer of fatty acid products per second at the interface. It was found that oleic acid rapidly dissolves into the oil phase under the conditions used. The interfacial tension measured reflects the adsorption of cutinase to the oil droplet and also responds to the fate of the hydrolysis products. A model is presented that describes the catalytic events at the oil-water interface during lipid hydrolysis. PMID- 9853366 TI - Characterization of cytotoxic factors of Yersinia pseudotuberculosis using the MDBK cell line. AB - The cytotoxin of four strains of Yersinia pseudotuberculosis was characterized using the MDBK cell line and by application of the MTT colorimetric test. The highest cytotoxin yield was obtained in tryptic soy broth medium after 24 h. It was detected in the cell-free culture filtrate, and treatment of the cells with CHAPS as a membrane detergent did not decrease significantly their cytotoxic activity. The cytotoxin was inhibited by trypsinization and by increasing values of either acidity or alkalinity. The cytotoxin was inactivated partially by heating at 70 degrees C for 20 min and totally at 90 degrees C for 10 min. The results obtained indicate that the cytotoxin is protein in nature and produced mainly as free exotoxin. PMID- 9853367 TI - Electrophoretic investigation on seven species of Indian frogs. AB - Studies on the occurrence of protein polymorphism are of importance not only in determining interspecific relationships but also in revealing genetically controlled variants among the populations of the same species. This study deals with the analysis of the isozyme pattern of three dehydrogenases in seven species of Indian frogs. The results correlate with the respective habitats and it is suggested that protein polymorphism is adaptively important and is maintained by natural selection. PMID- 9853368 TI - An alpha-actinin isoform which may cross-link intermediate filaments and microfilaments. AB - The G.3.5 antigen (named for the monoclonal antibody which recognizes it) has been characterized as an intermediate filament-associated protein found in a variety of tissue types, including human and rat astrocytes, rat skeletal and cardiac myocytes, fibroblasts, rat hepatocytes, and chicken and fish retinal tissues. Sequencing of proteolytic fragments indicated a high degree of similarity to alpha-actinin. Comparison of the G.3.5 antigen to alpha-actinin revealed that alpha-actinin and the G.3.5 antigen migrated similarly in reducing and non-reducing environments and had similar molecular masses (approximately 100,000). Overlay-immunoblotting assays indicated that the G.3.5 antigen and alpha-actinin could bind filamentous actin and desmin simultaneously. In contrast, immunocytochemistry indicated the G.3.5 antigen and alpha-actinin were immunologically distinct in tissue sections. The results of this study suggest that the G.3.5 antigen is an isoform of alpha-actinin which may serve to cross link intermediate filaments to microfilaments, and that other isoforms of alpha actinin may also share this property. PMID- 9853369 TI - A comparison between the use of a high-resolution CCD camera and 35 mm film for obtaining coloured micrographs. AB - In light microscopy, colour CCD cameras are now capable of generating image data sets that contain more information than can be captured with slow 35 mm colour reversal film. The resolution of colour CCD cameras with a high density of sensor elements (> or = 3300 x 2200 per channel of colour) is equivalent to that of slow 35 mm colour film over typical fields of view for objectives with a wide range of magnifications and numerical apertures. The contrast that can be achieved in images derived from the data sets obtained with colour CCD cameras far exceeds that found with film and can exceed that of human vision. Finally, the data sets collected with high-resolution colour CCD cameras are capable of being displayed at a wide range (four-fold) of different magnifications easily and interchangeably. Consequently, the combination of a data set that describes a relatively large field of view with one or two data sets that describe specific details taken with an eight-fold increase in magnification are all that is necessary to describe the salient features of the vast majority of stained specimens examined with transmitted light microscopy. PMID- 9853371 TI - Extended confocal microscopy of myocardial laminae and collagen network. AB - Ventricular myocardium has a complex three-dimensional structure which has previously been inferred from two-dimensional images. We describe a technique for imaging the 3D organization of myocytes in conjunction with the collagen network in extended blocks of myocardium. Rat hearts were fixed with Bouin's solution and perfusion-stained with picrosirius red. Transmural blocks from the left ventricular free wall were embedded in Agar 100 resin and mounted securely in an ultramicrotome chuck. Confocal fluorescence laser scanning microscopy was used to obtain 3D images to a depth of 60 microns in a contiguous mosaic across the surface. Approximately 50 microns was then cut off the surface of the block with an ultramicrotome. This sequence was repeated 20 times. Images were assembled and registered in 3D to form an extended volume 3800 x 800 x 800 microns 3 spanning the heart wall from epicardium to endocardium. Examples are given of how digital reslicing and volume rendering methods can be applied to the resulting dataset to provide quantitative structural information about the 3D organization of myocytes, extracellular collagen matrix and blood vessel network of the heart. PMID- 9853370 TI - An optical method for recording the activity of single transporters in membrane patches. AB - We have previously introduced an optical technique for recording the transport of fluorescent substrates by single membrane transporters. Referred to as optical single-transporter recording (OSTR), the method was restricted to cases in which membrane transporters occurred at extremely small densities, namely at one or a few transporters per cell. Here we describe the extension of OSCR from whole cells harbouring a small number of transporters to small membrane patches containing transporters at normal densities. A technique was developed for firmly attaching cells to isoporous filters, i.e. very thin transparent sheets containing homogeneous populations of cylindrical pores. The flux of fluorescent transport substrates across the tiny membrane pieces spanning the filter pores was measured by scanning microphotolysis, a combination of fluorescence microphotolysis and confocal laser scanning microscopy. The technique was tested by attaching erythrocytes to filters containing pores of 1.2, 2.0 or 3.0 microns diameter. After treating filter-attached erythrocyte membranes with streptolysin O, the transport of the fluorescent protein B-phycoerythrin through single streptolysin O pores was observed. From the flux data the functional radius of the streptolysin O pore was derived to be 12.5 +/- 0.9 nm, in very good agreement with previous electron microscopic estimates. The new technique features a number of unique properties: (i) the size of the membrane patch can be chosen within wide limits according to transporter density, (ii) transport can be recorded on many membrane patches in parallel, (iii) both influx and efflux may be analysed employing either photobleaching of fluorescent or photorelease of caged nonfluorescent substrates, (iv) two or more transport substrates may be monitored simultaneously. The new technique can be used, for instance, for analysing the activity of protein/particle pumps, a membrane transport domain not previously accessible to a single-transporter analysis. PMID- 9853372 TI - Measurement accuracy in confocal microscopy. AB - Confocal laser scanning microscopy provides optical serial sections through thick biological samples, making it possible to perform both three-dimensional visualization and three-dimensional quantitative analysis. On human lymphocytes, we measured geometrical features, cell contents in DNA and in cyclin A and CDK1 proteins, localization and colocalization of these two proteins. Cells were acquired at a vertical sampling step of 0.5 micron, which gives sufficient information about cell labelling. For the purpose of obtaining fast and reliable data at a reduced time cost, we examined various possibilities to simplify acquisition. For example, it might be possible to increase the vertical sampling step to 2.0 microns while preserving an acceptable accuracy of measurements. Further limiting the acquisition to the central sections appeared to give only rough estimations about the whole cells. Finally, we compared confocal microscopy to conventional two-dimensional epifluorescence microscopy. Confocal microscopy appeared slightly less accurate as regards content estimation, but was an invaluable tool when investigating three-dimensional structures and, more especially, localization of proteins. PMID- 9853373 TI - The coefficient of error of optical fractionator population size estimates: a computer simulation comparing three estimators. AB - The optical fractionator is a design-based two-stage systematic sampling method that is used to estimate the number of cells in a specified region of an organ when the population is too large to count exhaustively. The fractionator counts the cells found in optical disectors that have been systematically sampled in serial sections. Heretofore, evaluations of optical fractionator performance have been made by performing tests on actual tissue sections, but it is difficult to evaluate the coefficient of error (CE), i.e. the precision of a population size estimate, by using biological tissue samples because they do not permit a comparison of an estimated CE with the true CE. However, computer simulation does permit making such comparisons while avoiding the observational biases inherent in working with biological tissue. This study is the first instance in which computer simulation has been applied to population size estimation by the optical fractionator. We used computer simulation to evaluate the performance of three CE estimators. The estimated CEs were evaluated in tests of three types of non random cell population distribution and one random cell population distribution. The non-random population distributions varied by differences in 'intensity', i.e. the expected cell counts per disector, according to both section and disector location within the section. Two distributions were sinusoidal and one was linearly increasing; in all three there was a six-fold difference between the high and low intensities. The sinusoidal distributions produced either a peak or a depression of cell intensity at the centre of the simulated region. The linear cell intensity gradually increased from the beginning to the end of the region that contained the cells. The random population distribution had a constant intensity over the region. A 'test condition' was defined by its population distribution, the period between consecutive sampled sections and the spacing between consecutive sampled disectors. There were 1000 independently simulated cell populations for each test condition, and a 'trial' was conducted for each of these cell populations. In each trial we calculated the (unique) true CE of the population size estimate and the three CE estimates obtained by applying the Scheaffer-Mendenhall-Ott (SMO) and both Gundersen-Jensen (GJ) estimators. We compared the estimated CEs with the true CEs for each population distribution. We found that the CE estimates obtained by the SMO estimator were closer to the true CEs and had less scatter than those of the nugget-modified GJ estimator. Both had small positive bias. The CE estimates obtained by the unmodified GJ estimator exhibited widely varying bias and large scatter. In all the population distributions we tested, the average true CE was very nearly proportional to 1/square root of QT, where QT is the average number of cells counted in the two stage systematic sample. PMID- 9853374 TI - Electron and light microscopy studies on particulate EPR spin probes lithium phthalocyanine, fusinite and synthetic chars. AB - This is the first detailed study on the morphology of the widely used particulate electron paramagnetic resonance (EPR) probes lithium phthalocyanine (LiPc), methyl-LiPc, methoxy-LiPc, fusinite and synthetic carbon-based chars, by means of both light and electronmicroscopy (LM and EM). The importance of these EPR probes for the measurement of O2/NO has been reported previously. Under LM, LiPc with its distinct crystalline structure differs significantly from the noncrystalline black flakes of both LiPc derivatives, methyl-LiPc and methoxy-LiPc. Unlike the shiny carbon-based synthetic chars, which have no characteristic morphology, fusinite, a fraction of fossilized coal maceral, displays distinctive fine, parallel channels. SEM studies reveal a striated surface and interlocking multilayered structure of LiPc that is markedly different from either the multilayered stacked methyl-LiPc or- botryoidal methoxy-LiPc. The regularly spaced pores and channels of fusinite, a reflection of its plant origin, contrast sharply with the randomly distributed pores of all sizes of the various synthetic chars. Furthermore, the combined results of both LM and EM studies strongly suggest the nonperturbing nature of fusinite and LiPc when they were used as EPR probes to measure oxygen in cells or tissues. We hope that this study, in conjunction with the numerous EPR functional studies of probes reported earlier, provides valuable information for the future development of new particulate EPR spin probes. PMID- 9853375 TI - Towards automatic cell identification in DIC microscopy. AB - A general method is proposed for constructing templates of cells in differential interference contrast (DIC) microscopy. This takes account of the optics which generate DIC images, and is applicable to both transparent and semi-transparent cells of simple and complex shapes. Then, a template matching methodology is presented, which uses fast Fourier transforms to fit templates of a range of sizes and orientations to images. For illustration, this is used to automatically identify and measure individual Candida yeast cells in clusters. PMID- 9853376 TI - Electron beam-induced changes in vitreous sections of biological samples. AB - Nonpretreated high pressure frozen samples of Zea mays, cartilage and human erythrocytes were cryosectioned and observed at 110K in a cryoelectron microscope. Changes induced by medium doses of electron irradiation (< 10 ke nm 2) are described. After some ke nm-2, the most conspicuous cutting artefacts are erased to a large extent and the visibility of the cell organelles is improved. The sections, compressed in the cutting direction by the sectioning process, shrink once more, in the same direction, when irradiated. This shrinkage depends on the section support and on how the section is adsorbed to it. Shrinkage is not uniform: it is most pronounced in mitochondria, condensed chromatin and nucleolus. This differential shrinkage improves the visibility of major structures on the section and, as a result, 'nicer' images are recorded. However, this apparent improvement is a beam-induced artefact that must be paired with a loss of high resolution information. PMID- 9853377 TI - Rapid staining of ultrathin sections with the use of a microwave oven. AB - The microwave oven has many potential applications, ranging from tissue fixation to staining for light and electron microscopy. This study was planned to speed up the staining of ultrathin sections. The first set of grids was stained conventionally with uranyl acetate and Reynold's lead citrate solutions. The other grids were stained with the same solutions by microwave irradiation. The electron micrographs of grids stained using the microwave technique were as satisfactory as the grids stained conventionally. Microwave-treated grids demonstrated more uniform staining and less precipitate. The use of a microwave oven shortened staining time by approximately 38 min. PMID- 9853378 TI - Characterization of arylamine N-acetyltransferase in Enterobacter aerogenes. AB - N-acetyltransferase (NAT) activity was determined by incubation of purified Enterobacter aerogenes enzyme with 2-aminofluorene (2-AF) as the substrate, followed by high pressure liquid chromatography assays. The NAT activity from E. aerogenes was 0.58 +/- 0.08 nmol/min/mg protein for 2-AF. The values of apparent K(m) and Vmax were 0.72 +/- 0.14 mM and 2.45 +/- 0.29 nmol/min/mg protein, respectively, for 2-AF. The optimal pH value for the enzyme activity was 7.5 for the 2-AF tested. The optimal temperature for enzyme activity was 37 degrees C for the 2-AF substrate. The molecular weight of NAT from E. aerogenes was 44.9 kD. Among a series of divalent cations and salts, Zn2+, Ca2+, and Fe2+ were demonstrated to be the most potent protease inhibitors, and only ethylenediaminetetraacetic acid significantly protected the NAT. Iodoacetamide, in contrast to other agents, markedly inhibited NAT. PMID- 9853379 TI - HPLC analysis of quinolinic acid, a NAD biosynthesis intermediate, after fluorescence derivatization in an aqueous matrix. AB - Quinolinic acid (2,3-pyridine dicarboxylic acid), a biological intermediate in nicotinamide adenine dinucleotide (NAD) biosynthesis in microbes and mammals and a brain excitotoxin, is not fluorescent nor electrochemically active and its detection sensitivity by UV absorption is comparatively low. Quinolinic acid was successfully derivatized in water-based samples by monodansylcadaverine, a fluorescence tag, and analysed by high-performance liquid chromatography (HPLC). No extraction procedure was needed and quinolinic acid was activated by water soluble carbodiimide and derivatized under mild conditions. As little as 3 pmol (500 pg) of quinolinic acid in 5 microliter of artificial cerebrospinal fluid sample volume could be derivatized and detected at a signal to noise ratio of 3:1. Thus, detection on a mass basis by HPLC after fluorescence derivatization is about 300 times as sensitive as direct determination of quinolinic acid by UV absorbance (500 pg vs 150 ng). A variety of activators, fluorescent tags and reaction solvents and conditions were tested but found to be less effective. PMID- 9853380 TI - Evidence for an heterogeneous glycosylation of the Clostridium tyrobutyricum ATCC 25755 flagellin. AB - Glycosylation analysis of the flagellin from the Gram-positive species Clostridium tyrobutyricum has been supplemented. Amino acid analysis of the glycopeptides obtained after pronase digestion of flagellin indicated that O glycosylation which was previously demonstrated after nonreductive beta elimination, probably occurred via the hydroxyl group of serine. Otherwise, beta elimination partly deglycosylated flagellin. After this treatment carbohydrates were still linked to protein as shown by a digoxigenin-hydrazide labelling. Therefore, in addition to linkages via serine, alkaline resistant linkages exist on the flagellin and some glycans may be linked to the protein core via the amide nitrogen of asparagine or via the hydroxyl group of tyrosine. Furthermore, according to an immunological analysis, glycans attached to flagellin via alkaline sensitive linkages may be different from those attached via alkaline resistant linkages. PMID- 9853381 TI - A high resolving power ion selector for post-source decay measurements in a reflecting time-of-flight mass spectrometer. AB - An electrostatic deflector has been designed and constructed that can be used in a reflecting time-of-flight mass spectrometer for either single-deflector or dual deflector velocity selection in post-source decay measurements. The deflector consists of an interleaved set of parallel deflection electrodes as in a Loeb/Cravath/Bradbury device, but thin metal ribbon instead of wire is used for the deflection electrodes. The time for reversing the electric field, which depends on various factors such as the electronics for pulsing the voltage and the time constant of a particular electrode geometry, is about 19 ns for the deflectors used in this study. By properly timing the reversal of the electric field, the time-window for ion transmission can be made substantially less than the switching time of each individual deflector. In conjunction with matrix assisted laser desorption/ionization, the single-deflector's resolving power and transmission are robust with respect to laser fluence, i.e. they remain high even when the fluence is raised well above threshold. By contrast the operational features of the dual-deflector gate offer more versatility in locating and sizing the selection window. Operating the ion selector in a single-deflector mode, we have achieved a resolving power of approximately 710 full width at half maximum (FWHM) for different isotopes of protonated, sodiated, and potassiated substance P (m/z 1348.6, 1370.6 and 1386.6 respectively; 10.073 keV). Operating it in the dual-deflector mode under two different sets of conditions, we have succeeded in obtaining resolving powers of approximately 1100 (FWHM) for protonated substance P (m/z 1348.6; 10.8 keV) and approximately 5200 (FWHM) for an isotopomer of PEG 6000 (approximately m/z 6000; 10.04 keV). This accomplishment implies that high resolution ion selection can be coupled to post-source decay analyses. PMID- 9853383 TI - Monitoring the hybridization of the components of oligonucleotide mixtures to immobilized DNA via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Reported here is how the hybridization of individual components of oligonucleotide mixtures to solid-phase bound complementary strands can be monitored simultaneously by quantitative matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). Three oligonucleotides, a DNA heptamer, a DNA octamer and a DNA octamer with a terminal cholic acid appendage were used as the test mixture. Upon cooling in the presence of a complementary undecamer on controlled pore glass, depletion of the components from the solution was observed. The resulting hybridization curves show the same relative affinities as traditional UV melting curves with single components and their complement. Assays of the kind described here may be used to select high affinity binders from combinatorial libraries of modified antisense oligonucleotides. PMID- 9853382 TI - Mass spectrometric analysis of 2-deoxyribonucleoside and 2'-deoxyribonucleotide adducts with aldehydes derived from lipid peroxidation. AB - An important emerging issue in chemical carcinogenesis is the role that products of endogenous metabolism play in formation of covalently modified DNA. One example is the formation of alpha, beta-unsaturated aldehydes as a result of endogenous and drug-stimulated lipid peroxidation. Malondialdehyde (MDA), crotonaldehyde (CR), 2-hexenal (HX), and 4-hydroxy-2-nonenal (HNE) react covalently with 2'-deoxyguanosine (dG) and 2'-deoxyadenosine (dA) residues on DNA to form promutagenic cyclic adducts that may be important in the etiology of cancer in humans and animals. The accurate quantification of such adducts provides a powerful tool in molecular epidemiology for assessing carcinogenic risks from various lifestyle choices (e.g. diet, drug use) in humans. 32P Postlabeling is recognized as one of the most sensitive methods available for detection of DNA adducts in human tissues, but without adequate validation such methodology can yield inaccurate quantitative measurements. We have used LC separations in conjunction with electrospray ionization MS and tandem MS (triple quadrupole and hybrid quadrupole-orthogonal acceleration time of flight analyzers) to characterize MDA-, CR-, HX- and HNE-modified dG and nucleotide (3'- and 5'-monophosphate; 3',5'-bisphosphate) adducts. These data have been used to validate 32P-postlabeling methods for quantification of low level MDA-dG adducts formed in DNA of human and animal tissues. Availability of reliable methods for quantification of endogenous DNA damage in humans and animals is essential for determining unknown etiologies of cancer and for the assessment of cancer risks in humans. PMID- 9853384 TI - Determination of four anabolic steroid metabolites by gas chromatography/mass spectrometry with negative ion chemical ionization and tandem mass spectrometry. AB - A gas chromatography/mass spectrometry method is described which uses negative ion chemical ionization and tandem mass spectrometry for the determination of anabolic steroid metabolites. Four anabolic steroid metabolites to be derivatized by Pentafluoropropionic anhydride (PFPA) were determined using gas chromatography/mass spectrometry (GC/MS) with negative chemical ionization (NCI) and NCI/MS/MS. The repeatability and reproducibility of this procedure were in the range of 5.3-9.7% and 6.1-10.2%, respectively. This method of derivatization with PFPA for NCI and NCI/MS/MS was useful to determine four metabolites of nandrolone, dromostanolone, methenolone and boldenone. The derivatized metabolites of boldenone could be detected to 2 ppb and the other three steroids could be detected to 25 ppb in urine at a signal-to-noise ratio of S/N = 3. PMID- 9853385 TI - Identification of drug metabolites in biological matrices by intelligent automated liquid chromatography/tandem mass spectrometry. AB - A rapid and systematic strategy for the identification of drug metabolites in biological matrices based on liquid chromatography-tandem mass spectrometry (LC/MS/MS) techniques was utilized for the identification of drug metabolites of the HIV protease inhibitor Indinavir. This strategy integrates intelligent realtime mass spectrometry with HPLC detection and a predictive strategy for detecting metabolites arising from common biotransformations, to rapidly elucidate structures of drug metabolites. Structures of metabolites generated from in vitro incubation mixtures of Indinavir were characterized from a single chromatographic analysis using the automated LC/MS/MS methodology, thus reducing data acquisition time and improving efficiency. PMID- 9853386 TI - Identification of oxytocin and vasopressin from neurohypophyseal cell culture. AB - Our observation that dispersed cultures of neurohypophysis obtained from adult rats are capable of synthesizing and releasing oxytocin and vasopressin is unexpected, because in whole animals these hormones are known only to be stored, not to be produced in the posterior lobe of the pituitary. The hormone content of cell culture medium was elevated from 0 to 129 +/- 14 pg/mg protein for oxytocin and from 0 to 42 +/- 4 pg/mg protein for vasopressin during two weeks as determined by specific radioimmunoassay. By molecular mass and structure determination (tandem mass spectrometry) we have proved that the supernatant of the cell cultures contains not only immunologically but mass spectrometrically identified neurohypophyseal hormones. PMID- 9853387 TI - Structural analysis and quantitative evaluation of the modifications produced in human hemoglobin by methyl bromide using mass spectrometry and Edman degradation. AB - The present study reports a procedure developed for the identification and quantitative analysis of the adducts formed by interaction of methyl bromide with human hemoglobin, based on combined analysis by electrospray mass spectrometry and automated Edman degradation of either intact globin chains or tryptic peptides of globin chains. The procedure has allowed identification of the reactive sites in human hemoglobin, and has been applied to the analysis of samples modified in vitro by methyl bromide. The results obtained represent the basis for the complete structural characterization of the modified hemoglobin and demonstrate the usefulness of the proposed analytical approach for the evaluation of the degree of alkylation and the identification of modified amino acids in proteins. PMID- 9853388 TI - Identification of synaptic metabolites of dynorphin A (1-8) by electrospray ionization and tandem mass spectrometry. AB - Synaptic metabolism of the endogenous opioid octapeptide dynorphin (Dyn) A (1-8) (Tyr-Gly-Gly-Phe-Leu-Arg-Arg-Ile) was studied in vitro upon its incubation with synaptosomes and with synaptosomal plasma membranes isolated from rat brain tissue. Electrospray ionization (ESI) and tandem mass spectromety were performed using an ion trap instrument, and afforded the identification of Dyn A (2-8), Dyn A (1-6) and leucine enkephalin [Dyn A (1-5)] as major metabolites. Preliminary quantitative data on the kinetics of Dyn A (1-8) degradation and metabolite formation was obtained by size-exclusion chromatography/ESI tandem mass spectrometry, and revealed an apparent involvement of several enzymes in the metabolism upon incubation with synaptosomes. Predominant formation of Dyn A (1 6) was observed with the synaptosomal plasma membranes. PMID- 9853389 TI - Negative ion electrospray ionization tandem mass spectrometry in the structural characterization of penicillins. AB - The mass spectrometry (MS) behaviour of ten commercially available penicillins has been studied by means of electrospray and multiple-stage MS/MS experiments performed using an ion trap instrument. For all the examined compounds negative ions are produced under ESI conditions, with a yield two or three orders of magnitude higher than that observed for positive ions. MSn experiments indicate the occurrence of a fragmentation pathway related to the beta-lactam ring, different from that usually described for positive ions of these compounds, and provide structural information on both the beta-lactam ring and the side chains. PMID- 9853390 TI - Eyes closed: simple, intuitive, statistically sound, and efficient methods for estimating parameters of clonal growth cancer models. PMID- 9853391 TI - Rating threat mitigators: faith in experts, governments, and individuals themselves to create a safer world. AB - This research explores public judgments about the threat-reducing potential of experts, individual behavior, and government spending. The data are responses of a national sample of 1225 to mail surveys that include measures of several dimensions of public judgments about violent crime, automobile accidents, hazardous chemical waste, air pollution, water pollution, global warming, AIDS, heart disease, and cancer. Beliefs about who can best mitigate threats are specific to classes of threats. In general, there is little faith that experts can do much about violent crime and automobile accidents, moderate faith in their ability to address problems of global warming, and greater expectations for expert solutions to the remaining threats. People judge individual behavior as effective in reducing the threats of violent crime, AIDS, heart disease, and automobile accidents but less so for the remaining threats. Faith in more government spending is highest for AIDS and the other two health items, lowest for the trio of violent crime, automobile accidents, and global warming, and moderate for the remaining threats. For most threats, people are not distributed at the extremes in judging mitigators. Strong attitudinal and demographic cleavages are also lacking, although some interesting relationships occur. This relative lack of sharp cleavages and the generally moderate opinion indicate ample opportunity for public education and risk communication. PMID- 9853392 TI - A risk assessment for consumers of mourning doves. AB - Recreational and subsistence hunters and anglers consume a wide range of species, including birds, mammals, fish and shellfish, some of which represent significant exposure pathways for environmental toxic agents. This study focuses on the Department of Energy's (DOE's) Savannah River Site (SRS), a former nuclear weapons production facility in South Carolina. The potential risk of contaminant intake from consuming mourning doves (Zenaida macroura), the most popular United States game bird, was examined under various risk scenarios. For all of these scenarios we used the mean tissue concentration of six metals (lead, mercury, cadmium, selenium, chromium, manganese) and radiocesium, in doves collected on and near SRS. We also estimated risk to a child consuming doves that had the maximum contaminant level. We used the cancer slope factor for radiocesium, the Environmental Protection Agencies Uptake/Biokinetic model for lead, and published reference doses for the other metals. As a result of our risk assessments we recommend management of water levels in contaminated reservoirs so that lake bed sediments are not exposed to use by gamebirds and other terrestrial wildlife. Particularly, measures should be taken to insure that the hunting public does not have access to such a site. Our data also indicate that doves on popular hunting areas are exposed to excess lead, suggesting that banning lead shot for doves, as has been done for waterfowl, is desirable. PMID- 9853393 TI - Retrospective temporal and spatial mobility of adult Iowa women. AB - Human exposure assessments require a linkage between toxicant concentrations in occupied spaces and the receptor's mobility pattern. Databases reporting distinct populations' mobility in various parts of the home, time outside the home, and time in another building are scarce. Temporal longitudinal trends in these mobility patterns for specific age and gender groups are nonexistent. This paper describes subgroup trends in the spatial and temporal mobility patterns within the home, outside the home, and in another building for 619 Iowa females that occupied the same home for at least 20 years. The study found that the mean time spent at home for the participants ranged from a low of 69.4% for the 50-59 year age group to a high of 81.6% for the over 80-year-old age group. Participants who lived in either one- or two- story homes with basements spent the majority of their residential occupancy on the first story. Trends across age varied for other subgroups by number of children, education, and urban/rural status. Since all of these trends were nonlinear, they indicate that error exists when assuming a constant, such as a 75% home occupancy factor, which has been advocated by some researchers and agencies. In addition, while aggregate data, such as presented in this report, are more helpful in deriving risk estimates for population subgroups, they cannot supplant good individual-level data for determining risks. PMID- 9853394 TI - Sample timing and mathematical considerations for modeling breath elimination of volatile organic compounds. AB - Real-world exposure measurements are a necessary ingredient for subsequent detailed study of the risks from an environmental pollutant. For volatile organic compounds, researchers are applying exhaled breath analysis and the time dependence of concentrations as a noninvasive indicator of exposure, dose, and blood levels. To optimize the acquisition of such data, samples must be collected in a time frame suited to the needs of the mathematical model, within physical limitations of the equipment and subjects, and within logistical constraints. Additionally, one must consider the impact of measurement error on the eventual extraction of biologically and physiologically relevant parameters. Given a particular mathematical model for the elimination kinetics (in this case a very simple pharmacokinetic model based upon a multiterm exponential decay function that has been shown to fit real-world data extremely well), we investigated the effects on synthetic data caused by sample timing, random measurement error, and number of terms included in the model. This information generated a series of conditions for collecting samples and performing analyses dependent upon the eventual informational needs, and it provided an estimate of error associated with various choices and compromises. Though the work was geared specifically toward breath sampling, it is equally applicable to direct blood measurements in optimizing sampling strategy and improving the exposure assessment process. PMID- 9853395 TI - The efficacy of different methods for informing the public about the range dependency of magnetic fields from high voltage power lines. AB - The AC electric and magnetic fields associated with high voltage power lines have become a concern as a possible health risk. In most cases the strength of these fields decreases as the inverse square of the distance from the line. In earlier work, we found that laypeople do not understand how rapidly field strength decreases with distance. Most believe that any high voltage power line they can see is exposing them to strong fields. This paper confirms the earlier finding and explores a number of strategies which might be used in risk communications to correct this misperception. We found it relatively easy to provide subjects with a better understanding of the range-dependency of magnetic field strength. Moreover, the quality of this acquisition was apparently independent of the manner in which they were instructed. Such successful instruction is markedly different from the well-established difficulty of teaching people about many qualitative domains, such as physics or ideas in probability. Clearly, while some erroneous beliefs are highly resistant to change, others can be altered quite readily. We suspect that an important distinction between knowledge about the range-dependency of power-frequency magnetic fields and less tractable topics involves the presence or absence of prior folk-theories or "mental models" of the domain. PMID- 9853396 TI - A quantitative test of the cultural theory of risk perceptions: comparison with the psychometric paradigm. AB - This paper seeks to compare two frameworks which have been proposed to explain risk perceptions, namely, cultural theory and the psychometric paradigm. A structured questionnaire which incorporated elements from both approaches was administered to 129 residents of Norwich, England. The qualitative risk characteristics generated by the psychometric paradigm explained a far greater proportion of the variance in risk perceptions than cultural biases, though it should be borne in mind that the qualitative characteristics refer directly to risks whereas cultural biases are much more distant variables. Correlations between cultural biases and risk perceptions were very low, but the key point was that each cultural bias was associated with concern about distinct types of risks and that the pattern of responses was compatible with that predicted by cultural theory. The cultural approach also provided indicators for underlying beliefs regarding trust and the environment; beliefs which were consistent within each world view but divergent between them. An important drawback, however, was that the psychometric questionnaire could only allocate 32% of the respondents unequivocally to one of the four cultural types. The rest of the sample expressed several cultural biases simultaneously, or none at all. Cultural biases are therefore probably best interpreted as four extreme world views, and a mixture of qualitative and quantitative research methodologies would generate better insights into who might defend these views in what circumstances, whether there are only four mutually exclusive world views or not, and how these views are related to patterns of social solidarity, and judgments on institutional trust. PMID- 9853397 TI - Evaluating risk communication: examining target audience perceptions about four presentation formats for fish consumption health advisory information. AB - Information format can influence the extent to which target audiences understand and respond to risk-related information. This study examined four elements of risk information presentation format. Using printed materials, we examined target audience perceptions about: (a) reading level; (b) use of diagrams vs. text; (c) commanding versus cajoling tone; and (d) use of qualitative vs. quantitative information presented in a risk ladder. We used the risk communication topic of human health concerns related to eating noncommercial Great Lakes fish affected by chemical contaminants. Results from the comparisons of specific communication formats indicated that multiple formats are required to meet the needs of a significant percent of anglers for three of the four format types examined. Advisory text should be reviewed to ensure the reading level is geared to abilities of the target audience. For many audiences, a combination of qualitative and quantitative information, and a combination of diagrams and text may be most effective. For most audiences, a cajoling rather than commanding tone better provides them with the information they need to make a decision about fish consumption. Segmenting audiences regarding information needs and communication formats may help clarify which approaches to take with each audience. PMID- 9853398 TI - A wild-type Japanese measles virus strain inducing predominant early down regulation of CD46. AB - Down-regulation of CD46 secondary to stimulation with measles virus (MV) was investigated using CD46-positive cell lines and Japanese wild-type MV strains. The cells used were simian cell lines B95a and Vero in which MV strains have been adapted to be amplified, and Chinese hamster ovary (CHO) cell transfectants expressing human CD46 with (CHO(tail+)) or without (CHO(tail-)) the cytoplasmic tail. Of four Vero-adapted and three B95a-adapted MV strains, one Vero-adapted strain named Khono (KO), down-regulated CD46 within 60 min (early down regulation) in all cell lines examined except Vero. No strains other than Toyoshima (TY), which induced early down-regulation only in CHO(tail+) cells, induced early down-regulation of CD46 in any combination. On the other hand, conventional down-regulation of CD46 was observed 24 h post-MV inoculation (late down-regulation) when cell lines used were adapted to MV strains. Thus, we concluded that there are two modes of CD46 down-regulation by MV and the unique strain KO markedly induces early down-regulation. Also, the CD46 homologue of B95a, which fails to act as a MV receptor, is down-regulated concomitantly with MV replication (>24 h) in cells principally by competent virus strains. PMID- 9853399 TI - pH-dependent growth retardation of Escherichia coli caused by overproduction of Na+/H+ antiporter. AB - The overproduction of Na+/H+ antiporter NhaA in Escherichia coli caused growth retardation. Quantities and the activity of the antiporter were studied for their causative roles in terms of this retardation. We constructed a series of nhaA expression plasmids differing in their transcriptional and translational efficiencies. Low-level nhaA expression complemented the defect of an nhaA mutant and allowed the mutant to survive on a high-NaCl or high-LiCl medium. However, when the production of NhaA was strongly induced by the combination of a stronger promoter, an efficient translational initiation signal and a high copy number plasmid, the growth of the cells carrying the plasmid was severely retarded. This growth retardation correlated well with the amount of NhaA protein produced from the plasmids. Surprisingly, the growth retardation caused by overproduction of NhaA was enhanced more extensively at an alkaline pH than at a neutral pH, in which the antiporter activity was stimulated. However, these retardations were also observed for mutant NhaA antiporters without the activity. These results indicated that the growth retardation was due to an overproduction of the antiporter rather than its increased antiporter activity, and also affected by a pH-dependent change in NhaA, possibly its structural change. PMID- 9853400 TI - Effect of sulfated compounds on acid sialidase. AB - The inhibitory effects of various sulfated compounds on the activities of sialidases purified from porcine liver and human placenta were investigated. Among the sulfated compounds tested, heparin, dextran sulfate, condroitin sulfates and sulfatide significantly inhibited the 4-methylumbelliferyl-alpha-N acetylneuraminic acid (4-MU-NeuAc) sialidase activities of the two enzyme preparations, but glucose 6-sulfate and glucosamine 6-sulfate did not. Potassium sulfate showed an inhibitory effect only at high concentrations. When the sialidase activities were measured using natural substrates, the sialidase activities for the (alpha2-3) and (alpha2-6) sialyllactoses, and colominic acid, were markedly inhibited by heparin and sulfatide similar to 4-MU-NeuAc, although the fetuin sialidase activity was not significantly influenced by them. The sialidase activity hydrolyzing GM3 was strongly inhibited by heparin, but not by sulfatide. PMID- 9853401 TI - Identification of proteins whose amounts are altered by mutation in the pgsA gene of Escherichia coli. AB - We identified proteins whose amounts were altered in an Escherichia coli pgsA3 mutant lacking the potential to synthesize phosphatidylglycerolphosphate, a precursor of phosphatidylglycerol. Proteins whose amounts were increased in the mutant were protease Do, periplasmic oligopeptide-binding protein, tryptophanase, and an unidentified protein, while the decreased one was flagellin. Transformation of the mutant with a plasmid containing the wild type pgsA gene complemented the phenotype, indicating that the pgsA3 mutation is responsible for the phenotype. PMID- 9853402 TI - Comparison of the effects of gemfibrozil and clofibric acid on peroxisomal enzymes and cholesterol synthesis of rat hepatocytes. AB - We studied whether the peroxisomal proliferation, induction of 3-hydroxy-3 methylglutaryl-CoA reductase (HMG-CoA reductase) and activation of cholesterol synthesis by gemfibrozil shown in whole body (Hashimoto F., Ishikawa T., Hamada S. and Hayashi H., Biochemical. Pharm., 49, 1213-1221 (1995)) is also detected at a culture cell level, and we made a comparative analysis of the effects of clofibric acid. Gemfibrozil at 0.25 mM increased the activity of some peroxisomal enzymes (catalase and the cyanide-insensitive fatty acyl-CoA oxidizing system) after incubation for 72 h. However, contrary to whole body experiments, gemfibrozil decreased the activity of HMG-CoA reductase and cholesterol synthesis from [14C]acetate. At 1 mM, gemfibrozil decreased not only the activity of HMG CoA reductase and cholesterol synthesis, but also the protein content of the cells and peroxisomal enzyme activity, indicating nonspecific inhibition at this concentration. Clofibric acid (0.25 and 1 mM) increased the activity of peroxisomal enzymes, but decreased the activity of HMG-CoA reductase and cholesterol synthesis. With respect to the direct effect on HMG-CoA reductase in the cell homogenate, gemfibrozil at 0.25 mm did not affect the activity, but it clearly inhibited the activity at 2 mM and above. Clofibric acid at 2 mM hardly affected the activity, but it clearly decreased the activity at 5 mM and over. That is, gemfibrozil directly inhibited the activity more strongly than clofibric acid. The direct inhibition of the enzyme itself required higher concentrations of both agents than did inhibition at the culture cell level. These results suggest that the cytotoxicity of gemfibrozil is greater than that of clofibric acid, and that gemfibrozil, as well as clofibric acid, can induce peroxisomal enzymes in the culture cell level. In contrast to whole body results, gemfibrozil may suppress cholesterol synthesis from [14C]acetate through the inhibition of HMG-CoA reductase at the culture cell level. The decreases in the reductase activity caused by gemfibrozil and clofibric acid at the culture cell level may not be caused by the direct inhibition of the enzyme. PMID- 9853403 TI - Dual effects of anti-inflammatory 2-arylpropionic acid derivatives on a major isoform of human liver 3alpha-hydroxysteroid dehydrogenase. AB - Nonsteroidal anti-inflammatory drugs have been shown to be potent inhibitors of mammalian 3alpha-hydroxysteroid dehydrogenase. Here, we report that the drugs of the 2-arylpropionic acid class act as both activators and inhibitors for a predominant isoform of the human liver enzyme which is involved in the metabolism of steroid hormones, bile acids, drug ketones and xenobiotic aromatic hydrocarbons. Flurbiprofen, fenoprofen, ibuprofen, naproxen, ketoprofen and suprofen stimulated the activity of the human enzyme (1.5-2.4-fold) at low concentrations of less than 20-100 microM, whereas at higher concentrations they inhibited the activity. Comparison of the effects of the structurally related compounds with the drugs revealed that the essential structure required as the activator molecule is 2-phenylpropionic acid with a hydrophobic substituent on the aromatic ring. In addition, an R-enantiomer of ibuprofen showed higher activation (3-fold) than its S-enantiomer. Kinetic analysis with respect to NADP+ concentration indicated that R- and S-ibuprofens are nonessential activators showing binding constants of 23 and 34 microM, respectively. Neither enantiomers activated, but rather inhibited the enzyme, with Met replacing Arg-276 which has been shown to interact with the 2'-phosphate of NADP+. The inhibitions of the mutant enzyme by R- and S-ibuprofens were competitive with respect to the substrate, giving Ki values of 95 and 18 microM, respectively. The results suggest that the human liver 3alpha-hydroxysteroid dehydrogenase isoform possesses the two distinct sites, activator and inhibitor sites, to which anti inflammatory 2-arylpropionates stereoselectively bind. PMID- 9853404 TI - Anti-metastatic and immunomodulating properties of the water extract from Celosia argentea seeds. AB - We have investigated the anti-metastatic effect of Celosia argentea seed extracts (CAE), which have traditionally been used as a therapeutic drug for eye and hepatic diseases in China and Japan. Intraperitoneal (i.p.) administration of CAE for 7 d before tumor inoculation significantly inhibited liver metastasis caused by intraportal injection of colon 26-L5 carcinoma cells in a dose-dependent manner. CAE also showed concentration dependent mitogenic activity on BALB/c whole splenocytes, whereas incubation of the non-adherent fraction of splenocytes with CAE did not induce this activity. CAE has the ability to induce interleukin (IL)-12 production from macrophages in vitro. Following i.p. administration of CAE the maximal levels of IL-12 and interferon (IFN)-gamma production in serum were achieved at 2-3 and 6 h, respectively. Experiments using macrophage- or NK cell-deficient mice revealed that CAE-induced IL-12 in serum was not mediated by macrophages and that IFN-gamma production was mainly dependent on natural killer (NK) cells. Since CAE was inactive when the contributions of macrophages were removed in our system, its inhibitory mechanism is likely to be mainly associated with the activation of macrophages to an anti-metastatic state rather than NK cells. CAE administration resulted in increased production of IL-2, IFN-gamma and decreased production of a Th2 cytokine (IL-4) from splenocytes stimulated by PMA and A23187. Thus, the anti-metastatic effect by CAE is based on its immunomodulating properties including induction of cytokines such as IL-12, IL-2 and IFN-gamma leading to a Th1 dominant immune state and activating macrophages to the tumoricidal state. This may provide a basis for the inhibition of cancer metastasis. PMID- 9853405 TI - Phenobarbital-mediated induction of the CYP2B subfamily is not antagonized by picrotoxin, a potent antagonist of barbiturates in the central nervous system. AB - Our previous study indicated that picrotoxin competes with phenobarbital (PB) for the binding to liver microsomes, although the target of binding and the physiological role were not determined (unpublished observation). To seek information on the target site of PB, reflecting the induction of hepatic enzymes, we examined here the effect of picrotoxin on PB-mediated induction of hepatic cytochrome P450 and UDP-glucuronosyltransferase in vivo in rats. The induction of the CYP2B1/2 was estimated by immunoblot analysis and by measuring the activity of testosterone 16beta-hydroxylation. Intraperitoneal injection of picrotoxin alone slightly increased CYP2B1/2 protein. An experiment on co treatment of picrotoxin and PB showed that picrotoxin enhanced rather than antagonized the inducing effect of PB. The results suggest two possibilities: that 1) picrotoxin increases the CYP2B subfamily by binding to the same site as PB; or 2) the site in microsomes for the competition between PB and picrotoxin does not reflect the induction of P450. PMID- 9853406 TI - Vasoactive effects of cimicifugic acids C and D, and fukinolic acid in cimicifuga rhizome. AB - Vasoactive effects of cimicifugic acids A-E, fukinolic acid and fukiic acid isolated from Cimicifuga plants were investigated using rat aortic strips. Cimicifugic acid D and fukinolic acid at 3x10(-4) M caused a sustained, slowly developing relaxation of aortic strips precontracted with norepinephrine (NE) in preparations with or without endothelium. Cimicifugic acid C inversely caused a weak contraction. Cimicifugic acids A, B and E and fukiic acid showed no vasoactivity at 3x10(-4) M. Cimicifugic acids A-E and fukinolic acid are esters between cinnamic acids and the hydroxyl group of benzyltartaric acids. For the manifestation of vasoactivity in the rat aorta, it is concluded that in the cinnamic acid moiety, a caffeoyl group might be necessary for the relaxation activity, and the p-coumaroyl group causes contraction. Concentration response curves for the Ca2+-induced contracture of depolarized aortic strips with isotonic high K+ were not affected by cimicifugic acid D or fukinolic acid. The Ca2+-induced contraction of aortic strips, preincubated with 10(-6) M NE in the presence of 10(-6) M nicardipine and 0.01 mM EGTA in Ca2+-free solution, were inhibited by cimicifugic acid D and fukinolic acid. These results indicated that the inhibition by cimicifugic acid D and fukinolic acid of the NE-induced contraction of rat aorta are attributable to the suppression of Ca2+ influx from the extracellular space enhanced by NE. PMID- 9853407 TI - Pharmacological properties of traditional medicines. XXIV. Classification of antiasthmatics based on constitutional predispositions. AB - It is commonly accepted that bronchial asthma or rhinitis accompanies disorders in body fluids and body temperature. The effects of ephedrine and the traditional antiasthmatics "Makyo-kanseki-to" and "Goko-to" were therefore studied on such constitutional predispositions as insensible perspiration and body temperature. Ephedrine markedly increased body temperature and exhibited a strong increased action on respiratory insensible perspiration, whereas Makyo-kanseki-to and Goko to not only prevented the elevation of body temperature, but also increased respiratory insensible perspiration following the reduction of non-evaporative heat loss from the body surface. Thus, the diagnostic criteria of these two medicines used to treat hot-type asthma or dry cough were experimentally determined. The results also suggest that there is a great possibility that the administration of antiasthmatics may elicit side effects or make diseases worse unless their actions on constitutional predispositions are taken into account, such as body temperature and body fluids. PMID- 9853408 TI - Effects of paeoniflorin derivatives on scopolamine-induced amnesia using a passive avoidance task in mice; structure-activity relationship. AB - Paeoniflorin (1) and its derivatives having in common a cage-like pinane skeleton with hemiketal-acetal system, were evaluated for their effects on memory impairment induced by scopolamine in mice using a step-down type passive avoidance task. In the test session, 1 and its derivatives were intraperitoneally (i.p.) administered at doses of 0.002, 0.01, 0.02 and 0.2 mmol/kg, and 30 min later (15 min before the experiment), scopolamine (1 mg/kg, i.p.) was given. These compounds showed dose-dependent attenuation in a dose range of 0.002-0.02 mmol/kg and also enhancement of scopolamine-induced decrease in step-down latency. The effects of these compounds, except that of 2',3',4',5'-O-tetraacetyl 3-O-methylpaeoniflorin (8), followed a bell-shaped dose response profile. 8 Debenzoyl-6-deglucosyl-3-O-methylpaeoniflorin (6) showed no significant increase in the step-down latency at all tested doses. Maximum step-down latency was obtained by 3-O-methylpaeoniflorin (3) and 2',3,3',4',5'-penta-O methylpaeoniflorin (7) (the minimal effective dose was 0.002 mmol/kg). Relative to 3, debenzoylation, as in 8-debenzoyl-3-O-methylpaeoniflorin (4), slightly increased the latency, while deglucosylation, as in 6-deglucosyl-3-O methylpaeoniflorin (5), significantly reduced the prolongation of latency. Removal of both glucose and benzoyl moieties resulted in the loss of activity as seen in 6. These results revealed that, in addition to the cage-like pinane skeleton, the benzoyl and the glucosyl moieties are important structural elements of the paeoniflorin skeleton as its effects on scopolamine-induced amnesia. PMID- 9853409 TI - In vivo properties of the conjugates of mitomycin C with estradiol benzoate and estradiol: pharmacokinetics and antitumor characteristics against P388 leukemia and sarcoma 180. AB - Conjugates of mitomycin C (MMC) with estradiol benzoate and estradiol via glutaric acid, abbreviated to EB-glu-MMC and E-glu-MMC, respectively, as previously reported, were examined concerning their pharmacokinetic behaviors and antitumor effects against two kinds of general and popular tumors, P388 leukemia and Sarcoma 180. EB-glu-MMC and E-glu-MMC were dissolved in propylene glycol. Their solution was administered intraperitoneally to rats and mice in order to examine their plasma concentration-time profiles and antitumor characteristics, respectively. After the administration of EB-glu-MMC, EB-glu-MMC was detected slightly in blood only in the initial stage, while E-glu-MMC and MMC were observed there for a prolonged period. In the administration of E-glu-MMC, a similar phenomenon was observed but the drug retention effect seemed lower than that in EB-glu-MMC. In the antitumor test against P388 leukemia, E-glu-MMC exhibited a better effect than EB-glu-MMC; however, neither conjugate surpassed the effect of MMC. The toxic side effect was improved in each conjugate. As to the growth inhibition against Sarcoma 180, EB-glu-MMC and E-glu-MMC produced good effect and improved the toxic side effect. Especially, in the administration of EB-glu-MMC at the dose of 30 mg eq MMC/kg, a decrease in tumor volume was observed in the latter stage. EB-glu-MMC and E-glu-MMC were demonstrated to produce prolonged retention, to enter the systemic circulation to a fair extent, and to exhibit a good effect against the general solid tumor, Sarcoma 180, in vivo. PMID- 9853410 TI - Studies of drug delivery systems for a therapeutic agent used in osteoporosis. I. Pharmacodynamics (hypocalcemic effect) of elcatonin in rabbits following rectal administration of hollow-type suppositories containing elcatonin. AB - In this study, we developed a new hollow-type suppository containing elcatonin ((Asu1,7)-eel calcitonin, ECT), a synthetic derivative of eel calcitonin, which produces hypocalcemia, as a pharmaceutical preparation for self administration, to be used instead of parenteral injections for patients with osteoporosis. The absorption of ECT from the rectal mucous membrane was evaluated by observation of the decrease in serum calcium (Ca) concentrations following rectal administration in rabbits. ECT was efficiently absorbed from the rectum and effectively decreased serum Ca concentrations. The data of the area under the percent decrease in serum Ca concentration (deltaCa%)-time curve (deltaCa%-AUC), assumed to be an index of the pharmacodynamics (pharmacological effect) of ECT, indicated that similar hypocalcemic effects were obtained following rectal and intravenous administrations of ECT. In regard to the effect of coadministration of other compounds on rectal absorption of ECT, no significant difference in the deltaCa% AUC between rectal ECT administration with or without nafamostat mesilate (a protease inhibitor) was observed. However, the coadministration of ECT with cytochalasin B or monensin (endocytosis inhibitors) significantly decreased the deltaCa%-AUC, indicating that rectal ECT absorption is probably inhibited by endocytosis inhibitors. On the other hand, it was found that sodium decanoate, a medium-chain fatty acid (sodium salt), significantly enhanced the rectal absorption of ECT. We conclude that this ECT hollow-type suppository offers promise as a new method for the administration of ECT. PMID- 9853411 TI - Studies of drug delivery systems for a therapeutic agent used in osteoporosis. II. Enhanced absorption of elcatonin from nasal mucosa in rabbits. AB - In this study, the effects of a protease inhibitor, endocytosis inhibitors and an absorption-enhancing agent on the absorption of ((Asu1,7)-eel calcitonin, ECT) from the nasal mucous membrane in rabbits were examined, and the results were compared with those obtained following the rectal absorption of ECT reported in our previous paper. ECT was efficiently absorbed from the nasal mucous membrane and effectively decreased serum calcium (Ca) concentrations. The increase in the area under the percent decrease in serum Ca concentration (deltaCa%)-time curve (deltaCa%-AUC) value, assumed to be an index of the pharmacodynamics (hypocalcemic effect) of ECT, depended on the dose of ECT administered intranasally. When nafamostat mesilate, a protease inhibitor, was coadministered with ECT, the deltaCa%-AUC markedly increased. It is presumed that the influence (enzymatic barrier function) of protease on the nasal absorption of ECT is significant. However, no significant difference in the deltaCa%-AUC value was observed when an endocytosis inhibitor (cytochalasin B or monensin) was coadministered with ECT. ECT administration in combination with sodium decanoate, the sodium salt of a medium-chain fatty acid, effectively increased the deltaCa% AUC value due to the enhancing effect of sodium decanoate on the nasal absorption of ECT. We conclude that the nasal application offers a promising approach for the administration of pharmaceutical preparations containing ECT with additives such as nafamostat mesilate and sodium decanoate. PMID- 9853412 TI - Factors determining drug residence in skin during transdermal absorption: studies on beta-blocking agents. AB - The factors determining drug residence in skin during penetration across rat abdominal skin were investigated using five beta-blocking agents with different lipophilicities as model drugs in vivo and in vitro. The amount of beta-blocking agent in the skin at steady state correlated well with lipophilicity. The distribution of beta-blocking agents to the stratum corneum and the contribution of intercellular lipids in the stratum corneum to their skin distribution were also correlated with their lipophilicity, suggesting that the stratum corneum, especially intercellular lipids in the stratum corneum, would be responsible for the residence of beta-blocking agents in the skin. Furthermore, cholesterol-3 sulfate, palmitic acid, stearic acid and oleic acid were found to interact with the beta-blocking agents, which are cationized under the physiological condition, and were assumed to play an important role in the skin accumulation. On the other hand, the binding to keratinocyte was so small that keratinocyte might have little effect on the skin accumulation of the beta-blocking agents. Drug transport from the stratum corneum to viable skin was suggested to be regulated by the lipophilicity of these agents. To investigate the residence of these drugs in viable skin, in vitro transport studies using stripped skin were performed. The transport rate constant across viable skin to receptor cells (k23) was inversely correlated with the lipophilicity of the drugs. The elimination rate constants from viable skin (k(vs)) obtained in the in vivo study were much smaller than the values of k23 obtained in the in vitro study, and they were inversely correlated with the binding to cytosol components of viable skin but not the lipophilicity. The viable skin-to-muscle concentration ratio of these drugs, obtained at the beta-phase of the plasma concentration-time curve after intravenous administration, was also inversely correlated with the binding to the cytosol components of viable skin. These results suggest that k(vs) reflects the transport from viable skin to muscle rather than to blood circulation and that the binding of drugs to cytosol components in viable skin would be one of the important factors determining the residence in viable skin. PMID- 9853413 TI - Evaluation of poly(vinyl alcohol)-gel spheres containing chitosan as dosage form to control gastrointestinal transit time of drugs. AB - Two types of poly(vinyl alcohol)-gel spheres were prepared with chitosan (CS/PVA GS) and without chitosan (PVA-GS), and comparative studies were performed using these gel spheres (GSs). No change in particle size was observed by the addition of chitosan: nearly 45% of both particles were in the 5-10 microm range. In an in vivo gastrointestinal transit test, CS/PVA-GS prolonged the small-intestinal transit time more than PVA-GS. In an in vitro intestinal perfusion study, the mean transit time of these GSs was markedly reduced by pretreatment of the intestinal surface with a mucolytic agent, N-acetyl-L-cysteine, suggesting that the mucous layer on the intestinal surface plays an important role in controlling the transit rate of these GSs. The oral administration of aminophylline (theophylline) and ampicillin as model drugs incorporated in PVA-GS and CS/PVA-GS was examined in rats. While theophylline absorption from PVA-GS was not affected by the addition of chitosan, the improvement of ampicillin absorption by PVA-GS was enhanced by the chitosan combination. PMID- 9853414 TI - Simultaneous analysis of benzphetamine and its metabolites, and quantitation of urinary p-hydroxy-N-benzylamphetamine by micellar electrokinetic chromatography. AB - We developed a method for simultaneous analysis of benzphetamine (BZ) and its metabolites, p-hydroxy-N-benzylamphetamine (pHBA), p-hydroxybenzphetamine (pHBZ), amphetamine (AP), methamphetamine and p-hydroxymethamphetamine by micellar electrokinetic chromatography (MEKC). Urine samples from 0-15 h (3-h intervals) after oral administration of BZ (10 mg) were hydrolyzed with beta-glucuronidase (EC 3.2.1.31) at 37 degrees C overnight. The treated urine was applied to a solid phase extraction column Bond Elut Certify. After sequentially washing the column with water, 0.1 mol/l acetic acid and methanol, the samples were eluted with dichloromethane:isopropanol:28% ammonium hydroxide=78.4:19.6:2.0 (v/v %). The eluate was evaporated and the residue dissolved in running buffer was analyzed by MEKC. In urine from 0-3 h, AP, pHBZ and pHBA were detected. After that, only pHBA, which is one of the major metabolites of BZ in human urine, could be detected in the urine by the present method. A method for quantitation of pHBA by MEKC is described here. The effects of acetonitrile and sodium dodecyl sulfate in the running buffer of MEKC on the separation of BZ and its metabolites are also reported. PMID- 9853415 TI - Prevention of co-elution of steroid sulfates with serum proteins from pre-column in column-switching HPLC system. AB - A method to prevent co-elution of steroid sulfates with proteins in serum from the pre-column in column-switching HPLC was developed. The pre-column, a polymer coated mixed function column, was used for ion-pair chromatography with 5 mM tetra-n-butylammonium (TBA) ion. As steroid sulfates, estriol 3-sulfate, dehydroepiandrosterone 3-sulfate and pregnenolone 3-sulfate were used. Human serum (25 microl) was diluted with mobile phases including 5, 100 and 500 mM TBA ion, and then injected directly into the pre-column. The peak areas of the steroid sulfates in serum samples were compared with those of the steroid standards without serum. When 25/microl of serum was diluted with mobile phase including 100 or 500 mM TBA ion, the steroid sulfates in serum were retained in the pre-column; however, the steroid sulfates from the same sample diluted with mobile phase containing 5 mM TBA ion were not retained in the pre-column. Addition of an excess amount of counter ion (TBA ion) into the serum sample made it possible to retain the steroid sulfates in the pre-column. This method was applied to column-switching HPLC for measurement of steroid sulfates in serum using a semi-microcolumn as the analytical column. PMID- 9853416 TI - Evaluation of fluorescence polarization method for binding study in carbohydrate lectin interaction. AB - The fluorescence polarization (FP) technique was evaluated to determine molecular interaction between plant lectins and polysaccharides, yeast cells and glycopeptide after labeling the lectins with fluorescein isothiocyanate. Use of Lycoris radiata agglutinin allowed determination of the molecular interactions with large biomolecules containing mannose oligomers and polymers. Another example using a fluorescein-labeled glycopeptide also indicated that use of the FP method would allow easy observation of the molecular interactions on the quantitative base. The present technique is highly sensitive and facile because it does not require any washing procedures before measurement. PMID- 9853417 TI - Characterization of high- and low-molecular weight zinc-dependent acid phosphatases in bovine liver. AB - We have purified two forms of Zn2+-dependent acid phosphatase (Zn2+-APase) from bovine liver, both of which require Zn2+ to hydrolyze the substrate p-nitrophenyl phosphate in an acidic environment. The apparent molecular weights of these two forms of Zn2+-APase were estimated to be about 100,000 and 62,000 by gel filtration, and about 44,000 and 31,000 by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, respectively. The low-molecular weight (LMW) Zn2+-APase catalyzed the hydrolysis of myo-inositol-1-phosphate in the presence of 3 mM Mg2+ at physiological pH, but the high-molecular weight (HMW) enzyme did not. The LMW-Zn2+-APase of bovine liver was recognized by polyclonal antibodies developed against the Zn2+-APase of bovine brain, but the HMW-Zn2+ APase was not. PMID- 9853418 TI - Inhibitory effect of zinc and copper on phosphatase activity in the brain cytosol of rats: involvement of SH groups. AB - The effect of various metal ions on neutral phosphatase activity in the brain cytosol of rats was investigated. p-Nitrophenylphosphate was used as the substrate for the enzyme assay. Brain cytosolic phosphatase activity was significantly decreased by the presence of zinc or copper in the range of 10(-5) to 10(-3) M in the enzyme reaction mixture, while it was significantly increased by manganese, nickel or cobalt (10(-5) to 10(-3) M). Calcium (10(-5) to 10(-3) M) did not have an appreciable effect on brain cytosolic phosphatase activity. The inhibitory effect of zinc (10(-5) M) or copper (10(-5) M) on phosphatase activity was seen in the presence of manganese, nickel or cobalt with 10(-4) M concentration, indicating a potent inhibitory effect of these metals. Zinc or copper-decreased phosphatase activity was clearly prevented by dithiothreitol (5 mM), although the effect of each metal was further enhanced by N-ethylmaleimide (5 mM), suggesting that the metals act on the SH-groups of enzyme. Moreover, the inhibitory effect of zinc (10(-5) M) was significantly enhanced by S-100A (10(-6) M) or calbindin (10(-7) M), although neither protein had any effect in the presence of calcium (10(-5) M) or copper (10(-5) M). These results suggests that zinc has an unique role in the regulation of neutral phosphatase activity in rat brain cytosol. PMID- 9853419 TI - Studies of aloe. VI. Cathartic effect of isobarbaloin. AB - The cathartic effect of isobarbaloin, a stereoisomer of barbaloin (compound principally responsible for the cathartic activity of Aloe), was examined in male rats by oral administration. Individual differences in sensitivity in the laxative activity of isobarbaloin and barbaloin was not found. The cathartic activity (ED50) of isobarbaloin in barbaloin positive rats was 19.2 mg/kg, nearly equal to that of barbaloin (19.5 mg/kg). Also, isobarbaloin administered orally was demonstrated to decompose to aloe-emodin-9-anthrone (active metabolite of barbaloin) as well as to barbaloin. Therefore, it is considered that the mechanism underlying the cathartic effect of isobarbaloin is the same as that of barbaloin. PMID- 9853420 TI - An alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) antagonist and an N-methyl-D-aspartate (NMDA) channel blocker synergistically impair spatial memory in rats. AB - The interaction of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) antagonist YM90K with the competitive N-methyl-D-aspartate (NMDA) antagonist CGS19755 or with the NMDA channel blocker MK801 on the spatial memory of rats was examined by the 4-arm-baited radial maze task. When administered alone, these drugs had no effect on the spatial memory. The combination of YM90K and MK801 synergistically disrupted working and reference memories, whereas the combination of YM90K and CGS19755 had no effect. These results indicate that the blockade of the AMPA receptor and NMDA channels produces a synergistic impairment of spatial memory and suggest that interaction between the AMPA and NMDA receptors plays a role in cognitive function. PMID- 9853421 TI - Studies on Kochiae Fructus. V. Antipruritic effects of oleanolic acid glycosides and the structure-requirement. AB - We examined the antipruritic effects of various oleanolic acid glycosides from natural medicines such as Kochiae Fructus (the fruit of Kochia scoparia SCHRAD.) and Momordicae Radix (the roots of Momordica cochinchinensis SPRENG.) using a compound 48/80-induced pruritic model in mice. Oleanolic acid 3-O-monodesmosides showed an antipruritic effect, while oleanolic acid 3,28-O-bisdesmosides and their common sapogenol oleanolic acid lacked the activity. This evidence indicated that the 3-O-glycoside moiety and the 28-carboxyl group in oleanolic acid glycosides were essential for exhibiting the antipruritic effect. Furthermore, it was found that the 3-O-glucuronides showed more potent activity than the corresponding 3-O-glucosides. PMID- 9853422 TI - Structure-activity relationships of saponins and cardiac glycosides. III. Beta-L xylopyranosyl-(1-->6)-alpha- and -beta-D-glucopyranosides. AB - Comparisons of the biological activities of diosgenyl, methyl glycyrrhetinate or digitoxigenyl 3-O-beta-L-xylopyranosyl-(1-->6)-beta-D-glucopyranoside with those of other previously tested glycosides confirmed our assumption that both the hemolytic and antifungal activities of steroid saponins are generally parallel to each other, while almost all hemolytic triterpenoid saponins and nonhemolytic ones have no antifungal activity, and that cardiac diglycosides having a (1-->4) sugar linkage have stronger activities than those with a (1-->6) or a (1-->2) linkage. On the other hand, the case of the diosgenyl 3-O-beta-L-xylopyranosyl-(1 ->6)-alpha-D-glucopyranoside didn't conform to the above assumption, but those of methyl glycyrrhetinate and digitoxigenyl did. PMID- 9853423 TI - Change in erythrocyte shape induced by cyclosporine administration. AB - In order to estimate the effect of the long term administration of cyclosporine (CsA) on the shape change of erythrocytes, erythrocyte shapes which are observed with a scanning electron microscope were classified according to the nomenclature of Bessis for stomatocyte-echinocyte shape transformation. As a result of observing the erythrocyte shape of fifty-six patients with kidney transplantation treated with CsA, the morphological index of the erythrocytes of patients significantly increased to 0.0835+/-0.0085*** in comparison with 0.0004+/-0.0051 of those from healthy volunteers (control) (***: p<0.001, ANOVA). Such transformations had no relation to the subjects' sex or age. On the other hand, the erythrocytes of patients administered more than 100 ng/ml of CsA and posttransplanted within less than two years were transformed by CsA from the state of discocyte to echinocyte. In rats, the morphological index of erythrocytes of rats treated with 3 mg/kg/d or 5 mg/kg/d of CsA significantly increased in comparison with rats treated with saline (control). Furthermore, the erythrocytes of two patients were observed in terms of shape before the treatment with CsA. In both patients, the echinocyte type of erythrocyte increased by treatment with CsA. In vitro, the morphological index of the erythrocytes incubated with plasma containing CsA significantly increased, to 0.459+/-0.066*** in comparison with 0.064+/-0.029 of the control. It is suggested from these results that CsA treatment induces the echinocyte type of erythrocyte. PMID- 9853424 TI - Determination of insulin content in pancreatic beta cell line MIN6 cells by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. AB - The insulin content in mouse insulinoma MIN6 cells was determined using matrix assisted laser desorption ionization (MALDI) time-of-flight mass spectrometry (TOF/MS). A mass spectrum of cellular insulin was obtained with a cell burst of MIN6 by hypotonic water. Signal intensities of intracellular insulin were proportional to the number of MIN6 cells. The present method was applied to the determination of intracellular insulin content of MIN6 before and after glucose stimulation. PMID- 9853425 TI - Prognosis and prediction of response in breast cancer: the current role of the main biological markers. AB - In the medical literature there are frequently conflicting reports on the utility of biological tumour markers available in the clinical management of breast cancer. In this review we analyse current information on the relationships between the most widely investigated breast cancer biological markers including oestrogen and progesterone receptors, p53, Bcl-2, c-erbB-2, cyclin expression, proliferative activity, DNA ploidy and the urokinase plasminogen activation system, as well as their relevance to prognosis and response to clinical treatment. By biological prognostic indicator, we mean a marker that correlates with survival and disease-free survival; the term predictor marker indicates a marker that is capable of predicting tumour sensitivity or resistance to various therapies. Similarly to other authors' experiences, our analysis suggests that oestrogen receptors are weak prognostic indicators and good predictors of response to endocrine therapy. Furthermore, there are consistent data suggesting that proliferation indices are good indicators of prognosis, and that they are directly related to response to chemotherapy and closely related to response to hormonotherapy. On the contrary, there is no evidence or conflicting data for all of the other biological markers. These should be considered in the context of randomized trials in order to precisely define their prognostic and predictive roles. p53 and c-erbB-2 seem to be the most promising factors, but their use in routine practice still needs validation. PMID- 9853426 TI - Differences in C/EBPs in normal tissue and papillomas of the larynx. AB - Transcription factors belonging to the family of CAAT enhancer binding proteins (C/EBPs) programme differentiation in a wide variety of cells. We asked about the expression of C/EBPs in squamous epithelium. Using immunohistochemistry, C/EBPalpha and C/EBPbeta were easily detected in tissue sections from normal larynx or laryngeal papillomas, benign tumours with a papillomavirus aetiology and characterized by abnormal differentiation. A temporal expression of these C/EBPs occured as keratinocytes differentiated. In both tissues, C/EBPbeta appeared to be exclusively nuclear. In normal tissue, the greatest amount of C/EBPbeta was present in the spinous layer, and much less occurred in the granular layer. The basal layer of the laryngeal papillomas contained the most C/EBPbeta. Less was present in the spinous layer. Little C/EBPbeta was in the granular layer. Much C/EBPalpha was in the cytoplasm in both tissues. In normal tissue, nuclear C/EBPalpha staining was virtually absent in the basal layer, and became present in the spinous layer. Nuclear C/EBPalpha was randomly distributed in all layers in papillomas. Using immunoblots and Southwestern blots, we detected abundant truncated isoforms of C/EBPalpha in the papillomas. Since differentiation of many tissues is determined by the relative amounts of different C/EBPs, our data supports a role for C/EBPs in the differentiation of squamous epithelium. PMID- 9853427 TI - Prognostic significance of proliferative activity, DNA-ploidy, p53 and Ki-ras point mutations in colorectal liver metastases. AB - Paired colorectal liver metastases (CLM) and normal tissue samples from a consecutive series of 36 patients were studied prospectively. MIB-1 expression was studied by immunohistochemistry on paraffin-embedded sections. DNA ploidy and S-phase fraction (SPF) measurements were performed by flow cytometry on frozen tissues. Mutations within the p53 (exons 5-8) and c-Ki-ras (codons 12 and 13) genes were detected by PCR single-strand conformation polymorphism analysis followed by sequencing. A high correlation was observed between the MIB-1 LI and SPF value (rho=0.81; P<0.01). Moreover, p53 gene mutations were associated with either high MIB-1 LI and high SPF. In univariate analysis, SPF and MIB-1 levels were related to risk of death. The association between overall survival and DNA ploidy or p53 mutations did not reach statistical significance, but a slightly better survival was observed for patients either with DNA-diploid tumours or without mutations (P=0.05 and P=0.06, respectively). SPF was shown by multivariate Cox model analysis to be an independent prognostic variable and thus it might be a useful prognostic factor in patients with CLM. PMID- 9853428 TI - Co-ordination between localized wound-induced Ca2+ signals and pre-wound serum signals is required for proliferation after mechanical injury. AB - The signals which initiate proliferation of endothelial cells after injury are important for selective blood vessel growth during wound healing or tumour growth. Upon mechanically wounding quiescent cells, a transient [Ca2+]i increase was induced in cells at the wound edge. These same cells proliferated 18-24 h post wounding, as measured by bromodeoxyuridine incorporation. The localized Ca2+ signal was required specifically during wounding since blocking Ca2+ influx reduced proliferation by 40-50%. Proliferation also required serum since starvation reduced proliferation by 80%. Serum-starved cells proliferated if briefly primed with serum prior to wounding. The signals derived from serum and [Ca2+]i combined at least additively to induce proliferation. Therefore, serum priming followed by a single, transient Ca2+ signal induced by mechanical injury must occur in a temporally and spatially regulated manner for normal proliferation. Co-ordination between signalling cascades induced by growth factors and release from contact inhibition might be obligatory for localized re endothelialization after injury. PMID- 9853429 TI - Detention of potentially dangerous people. PMID- 9853430 TI - Facts, at last, on management of small infrarenal aortic aneurysms. PMID- 9853431 TI - Excitatory neurons and schizophrenia. PMID- 9853432 TI - Molecular epidemiology of familial hypercholesterolaemia. PMID- 9853433 TI - Substance-P antagonists: a new treatment for depression? PMID- 9853434 TI - Search for pathophysiology of panic disorder. PMID- 9853435 TI - Implications of genetic testing for insurance in the UK. PMID- 9853436 TI - Mortality results for randomised controlled trial of early elective surgery or ultrasonographic surveillance for small abdominal aortic aneurysms. The UK Small Aneurysm Trial Participants. AB - BACKGROUND: Early elective surgery may prevent rupture of abdominal aortic aneurysms, but mortality is 5-6%. The risk of rupture seems to be low for aneurysms smaller than 5 cm. We investigated whether prophylactic open surgery decreased long-term mortality risks for small aneurysms. METHODS: We randomly assigned 1090 patients aged 60-76 years, with symptomless abdominal aortic aneurysms 4.0-5.5 cm in diameter to undergo early elective open surgery (n=563) or ultrasonographic surveillance (n=527). Patients were followed up for a mean of 4.6 years. If the diameter of aneurysms in the surveillance group exceeded 5.5 cm, surgical repair was recommended. The primary endpoint was death. Mortality analyses were done by intention to treat. FINDINGS: The two groups had similar cardiovascular risk factors at baseline. 93% of patients adhered to the assigned treatment. 309 patients died during follow-up. The overall hazard ratio for all cause mortality in the early-surgery group compared with the surveillance group was 0.94 (95% CI 0.75-1.17, p=0.56). The 30-day operative mortality in the early surgery group was 5.8%, which led to a survival disadvantage for these patients early in the trial. Mortality did not differ significantly between groups at 2 years, 4 years, or 6 years. Age, sex, or initial aneurysm size did not modify the overall hazard ratio. INTERPRETATION: Ultrasonographic surveillance for small abdominal aortic aneurysms is safe, and early surgery does not provide a long term survival advantage. Our results do not support a policy of open surgical repair for abdominal aortic aneurysms of 4.0-5.5 cm in diameter. PMID- 9853437 TI - Health service costs and quality of life for early elective surgery or ultrasonographic surveillance for small abdominal aortic aneurysms. UK Small Aneurysm Trial Participants. AB - BACKGROUND: Uncertainty surrounds the best approach to management of patients with symptomless abdominal aortic aneurysms of 4.0-5.5 cm in diameter. The UK Small Aneurysm Trial showed that surveillance and early prophylactic elective surgical repair had similar survival benefits. We compared the impact on health service costs and quality of life of these two management strategies. METHODS: We measured and valued direct health service costs for use of National Health Service resources. We used the Medical Outcomes Study short-form patients' health survey to measure health-related quality of life in several areas of functional status and well-being. We did analyses by intention to treat for all 1090 patients in the 93 trial centres. FINDINGS: The mean cost of treatment in the early-surgery group was significantly higher than that for ultrasonographic surveillance (UK pounds sterling 4978 vs 3914, difference pounds sterling 1064 [95% CI 799-1328]). This finding was robust for a range of assumptions that varied the time between surveillance visits and the mean unit cost of elective aneurysm repair. Health-related quality of life was generally similar 12 months after randomisation for the two groups, but early-surgery patients reported positive improvement in current health perceptions and less negative change in bodily pain. INTERPRETATION: Cost was higher for early surgery than for ultrasonographic surveillance for small aortic aneurysms. Early surgery is, however, associated with improvement in some features of health-related quality of life that should be taken into account with the finding that early surgical repair gives no significant survival advantage over surveillance. PMID- 9853438 TI - Randomised placebo-controlled trial of use of topical recombinant bovine basic fibroblast growth factor for second-degree burns. AB - BACKGROUND: Wound healing is a dynamic process that could be accelerated by growth factors. We investigated the effect of recombinant bovine basic fibroblast growth factor (rbFGF) on burn healing in a randomised placebo-controlled trial. METHODS: We recruited 600 patients with superficial or deep second-degree burns. Patients received 150 AU/cm2 daily topical rbFGF (n=300) or placebo (n=300) plus vehicle. We assessed healing by photography, punch-biopsy, and clinical examination. FINDINGS: All patients treated with rbFGF had faster granulation tissue formation and epidermal regeneration than those in the placebo group. Superficial and deep second-degree burns treated with rbFGF healed in a mean of 9.9 (SD 2.5) days and 17.0 (4.6) days, respectively, compared with 12.4 (2.7) and 21.2 (4.9) days (p=0.0008 and p=0.0003, respectively). No adverse effects were seen locally or systemically with rbFGF. INTERPRETATION: rbFGF effectively decreased healing time and improved healing quality. Clinical benefits would be shorter hospital stays and the patient's skin quickly becoming available for harvesting and grafting. PMID- 9853439 TI - Differences in sexual risk behaviour between young men and women travelling abroad from the UK. AB - BACKGROUND: Identification of people who most frequently engage in sexual risk behaviour while travelling abroad would be useful for the design and targeting of health education and promotion campaigns. METHODS: Eligible participants were people living in the UK aged 18-34 years who had travelled abroad without a partner in the previous 2 years. Respondents were first screened for eligibility as part of representative face-to-face and telephone surveys by a market research company. Eligible individuals who agreed to take part then underwent a computer assisted telephone interview. Reinterviewing continued until 400 eligible people had been contacted. We also interviewed a control group of 568 young people who had travelled abroad without a partner in the previous 2 years but who did not report a new sexual relationship during their travels. FINDINGS: One in ten of the eligible participants reported sexual intercourse with a new partner. Travellers who reported a new sexual relationship abroad were also likely to report large numbers of sexual partners at home. Of the 400 people who had a new sexual partner abroad, 300 (75%) used condoms on all occasions with the new partner. Logistic regression modelling showed differences between men and women in those factors linked to the practice of unsafe or safer sex while travelling. For men, patterns of condom use abroad with casual partners (p<.0001) reflected patterns of use at home (p<0.001), whereas for women, patterns of condom use varied according to their partners' backgrounds (p<.0001). INTERPRETATION: Condoms are widely used among young travellers, but patterns of use vary by sex. Campaigns about sexual health targeted at international travellers should continue, not least because young people who meet new sexual partners abroad may be a convenient proxy group for that minority of the population who report most sexual partners at home. Such campaigns should be designed differently for men and women. PMID- 9853440 TI - Preferential involvement of excitatory neurons in medial temporal lobe in schizophrenia. AB - BACKGROUND: The anatomical basis of schizophrenia involves the cytoarchitecture of the cerebral cortex, but the phenotype of the affected neurons and synapses remains unclear. In mice, the presynaptic protein complexin I is a marker of axosomatic (inhibitory) synapses, whereas complexin II is a marker of axodendritic (mainly excitatory) synapses. These findings suggest that the complexins might be useful in the investigation of the synaptic pathology of schizophrenia. METHODS: We characterised the expression of the complexins in tissue taken at necropsy from human medial temporal lobe (hippocampus, parahippocampal gyrus) and cerebellum using in-situ hybridisation and immunoautoradiography. We then measured the concentrations of the complexins and their messenger RNAs (mRNAs) in the medial temporal lobe of 11 patients with schizophrenia and 11 non-schizophrenic controls. FINDINGS: The distribution of complexin I and II was consistent with the data on mice, with predominant expression of complexin I by inhibitory neurons, and complexin II by excitatory neurons. The amounts of both complexin mRNAs were lower in schizophrenic than in control patients (p<0.001), but the difference of complexin II mRNA was greater. The amount of complexin I protein was unchanged in schizophrenia, but complexin II protein was decreased (p<0.001). For both mRNA and protein, the complexin II/complexin I ratio was lower in schizophrenia, confirming the relatively greater loss of the excitatory marker. The findings did not seem attributable to medication. INTERPRETATION: The synaptic pathology of schizophrenia, at least in medial temporal lobe, primarily affects excitatory (glutamatergic) neurons. The inferred imbalance between excitatory and inhibitory circuitry may contribute to the involvement of this region in the pathophysiology of the disorder. PMID- 9853441 TI - Cranial-nerve palsies and vomiting. PMID- 9853443 TI - Anticipation in familial cavernous angioma: a study of 52 families from International Familial Cavernous Angioma Study. IFCAS Group. PMID- 9853442 TI - Successful fetal surgery for spina bifida. PMID- 9853444 TI - Adverse effect of blood group ABO mismatching on corneal epithelial cells. PMID- 9853445 TI - Prediction of cognitive decline in early Alzheimer's disease. PMID- 9853446 TI - L5-S1 disc origin for a pyramidal syndrome? PMID- 9853447 TI - Oral contraceptive intake in women with focal nodular hyperplasia of the liver. PMID- 9853448 TI - Ambient temperatures and potassium concentrations. PMID- 9853449 TI - Follow-up of patients with chronic anal fissure treated with topical glyceryl trinitrate. PMID- 9853450 TI - Detection of Mycobacterium tuberculosis in nasopharyngeal aspirate samples in children. PMID- 9853451 TI - Bartonella henselae infection from a dog. PMID- 9853452 TI - Chemoprevention controversies continue. PMID- 9853453 TI - HAART and host: balancing the response to HIV-1. Highly active antiretroviral therapy. PMID- 9853454 TI - Dodging bullets while searching for health care in Kosovo. PMID- 9853455 TI - Ghost of AIDS musical returns to haunt South Africa's health minister. PMID- 9853456 TI - Kidney cancer. AB - In the USA, the incidence of kidney cancer has increased 43% since 1973. The risk of the disorder is higher in men than in women and increases with age. The von Hippel-Lindau tumour-suppressor gene is inactivated in over 75% of sporadic cases. Metastatic disease is present in 20-30% of patients at diagnosis. Early stage kidney cancer is treated with a radical nephrectomy, but under certain circumstances a partial nephrectomy may be done. Tumour thrombus into the vena cava or right atrium requires thoracotomy and hypothermic circulatory arrest for successful removal of the tumour, but should not be done if extensive nodal or frank metastatic disease is present. Interleukin-2 is the systemic therapy of choice for metastatic disease at present, with long-term relapse-free survival of 5-8%. Several treatments including anti-angiogenesis drugs, cyclin-dependent kinase inhibitors, and differentiating agents are being actively investigated. Fluorouracil has a 10-15% response rate, and surgical excision of isolated metastases should always be considered. Therapy for metastatic renal cancer remains inadequate, but recent developments in basic and clinical research suggest future improvement. PMID- 9853457 TI - The discovery of heroin. PMID- 9853458 TI - The arterial organ in cardiovascular disease: ADAPT (arterial disease assessment, prevention, and treatment) clinic. PMID- 9853459 TI - Long-chain polyunsaturated fatty acids, infant formula, and breastfeeding. PMID- 9853460 TI - Long-chain polyunsaturated fatty acids, infant formula, and breastfeeding. PMID- 9853461 TI - Breastfeeding and maternal employment. PMID- 9853462 TI - Landfill sites and congenital abnormalities. PMID- 9853463 TI - Diagnosis of orthostatic hypotension. PMID- 9853464 TI - Socioeconomic status, standard of living, and neurotic disorder. PMID- 9853465 TI - Integration of human herpesvirus 6 genome in human chromosomes. PMID- 9853466 TI - Trends in prescribing calcium-channel blockers. PMID- 9853467 TI - Losartan and anaemia of end-stage renal disease. PMID- 9853468 TI - Imported epidemic typhus. PMID- 9853469 TI - Beta2 agonists and heart failure. PMID- 9853470 TI - Adverse effects of human insulin. PMID- 9853471 TI - Simultaneous infection of immunocompetent individuals with multiple cytomegalovirus strains. PMID- 9853472 TI - Framingham at 50. PMID- 9853473 TI - European breast-cancer meeting. PMID- 9853474 TI - Diabetes and renal cholesterol emboli. PMID- 9853475 TI - Hepatitis treatment in developing countries. PMID- 9853476 TI - Racism in the medical profession. PMID- 9853477 TI - Dengue shock syndrome and acute respiratory distress syndrome. PMID- 9853478 TI - Paul Ehrlich's ingredients for success. PMID- 9853479 TI - Exploring medical genetics on the web. PMID- 9853480 TI - The Nobel chronicles. 1927: Julius Wagner-Jauregg (1857-1940). PMID- 9853481 TI - The control of new prosthetic implants. PMID- 9853482 TI - Neonatal detection of developmental dysplasia of the hip (DDH) PMID- 9853483 TI - The Burch-Schneider anti-protrusio cage in revision total hip arthroplasty: indications, principles and long-term results. AB - Patients who had a revision total hip arthroplasty using the Burch-Schneider anti protrusio cage (APC) by a single surgeon have been reviewed after a minimum of five years. There were 63 operations in 58 patients with an average age of 63 years (41 to 83) at the time of revision. At an average follow-up of 8.5 years (5 to 18), 15 patients (25.9%) rated their results as excellent, 38 (65.5%) as good, and five (8.6%) as fair. Five further revisions of the acetabular prosthesis were required, three due to aseptic loosening, one for recurrent dislocation and one due to sepsis. Of the remainder, one was definitely loose, two probably loose, and 12 possibly so. Impressive augmentation of bone stock can be achieved with the anti-protrusio cage, while enabling the hip to be centred in its anatomical position. PMID- 9853484 TI - Estimation of the extent of osteonecrosis of the femoral head using MRI. AB - The extent of necrosis is the main determining factor in the outcome of osteonecrosis of the femoral head and is best measured by MRI. Using statistical analyses, we have evaluated the accuracy and repeatability of seven different methods of measuring the extent of necrosis, of which three demonstrate good or acceptable accuracy. For the general observer their repeatability is poor; only for experienced observers were they reasonable. We conclude that within an institution or for personal use, the accuracy of some methods is acceptable, and repeatability reasonable. None is appropriate for international use. PMID- 9853485 TI - Medium-term results of percutaneous, arthroscopically-assisted osteosynthesis of fractures of the tibial plateau. AB - We carried out percutaneous, arthroscopically- and fluoroscopically-assisted osteosynthesis of fractures of the tibial plateau in 52 patients, of whom 38 were assessed using the HSS knee score and standing radiographs. We reviewed 31 AO type-B fractures and seven type-C fractures after a mean follow-up of five years (1 to 14). Fixation was achieved using percutaneous screws and/or an external frame; 33 associated intra-articular injuries, diagnosed in 21 out of the 38 patients, were treated arthroscopically. Subjectively, 94.7% of the patients reviewed were satisfied. According to the HSS knee score 78.9% of the results were excellent, 13.2% good, 7.9% fair and none was poor. Narrowing of the joint space was found in 28.9% of the injured and 5.3% of the unaffected knees and axial deviation of 5 degrees to 10 degrees in 15.8% of the injured and 10.5% of the unaffected knees. Of the 52 fractures, reduction was incomplete in one, and in two secondary displacement occurred, of which one required corrective osteotomy. Deep-venous thrombosis occurred in four cases. The technique has proved to be safe but demanding. It facilitates diagnosis and appropriate treatment of associated intra-articular lesions. PMID- 9853486 TI - Treatment of painful articular cartilage defects of the patella by carbon-fibre implants. AB - Between 1989 and 1993 we implanted a matrix support prosthesis made of carbon fibre for the treatment of chronic painful articular defects of the patella in 27 patients. The mean period of follow-up was 33 months (11 to 54) with clinical and arthroscopic evaluation of the implant. Overall, there were four excellent, three good, seven fair and 13 poor results. Nine patients subsequently had a patellectomy for persistent pain at a mean of 27 months after surgery (14 to 47). The mean visual analogue pain scores in those who retained their patella were 7.6 before operation and 5.5 at the time of the latest follow-up. Patient satisfaction for the entire group was 41%. There appeared to be good incorporation of the prosthesis and a satisfactory ingrowth of fibrocartilage, but the poor results in 48% and low patient satisfaction discourage us from recommending the procedure for lesions of the articular cartilage of the patella. The consistent seeding of the joint with carbon-fibre debris and a histiocytic giant-cell reaction in the synovium are also a cause for concern. PMID- 9853487 TI - Randomised, prospective study comparing cemented and cementless total knee replacement: results of press-fit condylar total knee replacement at five years. AB - Early implants for total knee replacement were fixed to bone with cement. No firm scientific reason has been given for the introduction of cementless knee replacement and the long-term survivorship of such implants has not shown any advantage over cemented forms. In a randomised, prospective study we have compared cemented and uncemented total knee replacement and report the results of 139 prostheses at five years. Outcome was assessed both clinically by independent examination using the Nottingham knee score and radiologically using the Knee Society scoring system. Independent statistical analysis of the data showed no significant difference between cemented and cementless fixation for pain, mobility or movement. There was no difference in the radiological alignment at five years, but there was a notable disparity in the radiolucent line score. With cemented fixation there was a significantly greater number of radiolucent lines on anteroposterior radiographs of the tibia and lateral radiographs of the femur. At five years, our clinical results would not support the use of the more expensive cementless fixation whereas the radiological results are of unknown significance. Longer follow-up will determine any changes in the results and conclusions. PMID- 9853488 TI - Wear of congruent meniscal bearings in unicompartmental knee arthroplasty: a retrieval study of 16 specimens. AB - Many designs of unicompartmental knee replacement show early and mid-term failure due to polyethylene wear. We studied the wear rate of congruent polyethylene meniscal bearings retrieved from failed Oxford unicompartmental knee replacements. We examined 16 bearings, 0.8 to 12.8 years after implantation, measuring their thickness and comparing it with that of 14 unused bearings. The mean rate of penetration, which included the effects of wear at both upper and lower surfaces, was 0.036 mm per year (maximum 0.08). Bearings as thin as 3.5 mm wore no faster than thicker models, but ten with evidence of impingement had greater wear. The six bearings with no impingement showed a mean rate of penetration of 0.01 mm per year. In unicompartmental knee replacement, careful implantation of fully congruous meniscal bearings can avoid failure due to polyethylene wear. PMID- 9853489 TI - The Oxford medial unicompartmental arthroplasty: a ten-year survival study. AB - Retrieval studies have shown that the use of fully congruent meniscal bearings reduces wear in knee replacements. We report the outcome of 143 knees with anteromedial osteoarthritis and normal anterior cruciate ligaments treated by unicompartmental arthroplasty using fully congruous mobile polyethylene bearings. At review, 34 knees were in patients who had died and 109 were in those who were still living. The mean elapsed time since operation was 7.6 years (maximum 13.8). We established the status of all but one knee. There had been five revision operations giving a cumulative prosthetic survival rate at ten years (33 knees at risk) of 98% (95% CI 93% to 100%). Considering the knee lost to follow-up as a failure, the 'worst-case' survival rate was 97%. No failures were due to polyethylene wear or aseptic loosening of the tibial component. One bearing which dislocated at four years was reduced by closed manipulation. The ten-year survival rate is the best of those reported for unicompartmental arthroplasty and not significantly different from the best rates for total knee replacement. PMID- 9853490 TI - Expansion of an osteoarthritic cyst associated with wear debris: a case report. AB - We present a case in which the growth of an intraosseous cyst arising from the proximal tibiofibular joint appeared to have been increased by polyethylene wear particles from a medial unicompartmental knee replacement. Histological examination of the cyst wall showed a histiocytic response associated with numerous polyethylene wear particles. This case demonstrates that there is a direct communication between the joint cavity and the cyst. Such communication is probably through openings in the articular cartilage large enough to allow the passage of these particles. PMID- 9853491 TI - A technique for enhancing union of allograft to host bone. AB - The aim of limb-salvage surgery in malignant bone tumours in children is to restore function and eradicate local disease with as little morbidity as possible. Allografts are associated with a high rate of complications, particularly malunion at the allograft-host junction. We describe a simple technique which enhances union of allograft to host bone taking advantage of the discrepancy in size between the adult allograft and the child's bone. This involves lifting a flap of periosteum before resection from the host bone, which is then telescoped into the allograft medullary canal, which may require internal burring or splitting, for a distance of 1.5 to 2 cm and covering the bone junction with the periosteal flap. This is more stable than conventional end-to end opposition. For each centimetre of telescoping the surface area available for bony union is increased more than three times. The periosteal flap also augments union. Additional surface fixation with a plate and screws is not necessary. We have used this technique in nine children, in eight of whom there was complete union at a mean of 16 weeks. Delayed union, associated with generalised limb osteoporosis, occurred in one. Early mobilisation, with weight-bearing by three weeks, was possible. There was only one fracture of the allograft. PMID- 9853492 TI - Surgical stabilisation of the lower limb in osteogenesis imperfecta using the Sheffield Telescopic Intramedullary Rod System. AB - The Sheffield Expanding Intramedullary Rod System was developed after experiencing problems with existing rod systems in the management of osteogenesis imperfecta. Between 1986 and 1996 we treated 74 bones in the lower limb in 28 children at a median follow-up of 5.25 years. We have reviewed 24 children with a total of 60 rods. Before surgery, all children had had multiple fractures of the lower limb. At review eight patients had experienced no further fractures, but three had suffered five or more subsequently. Before initial stabilisation, 15 children had never walked, and only three (13%) used walking as their main means of mobility. After surgery, half of those who showed motor arrest were able to walk (p = 0.016). The number of patients able to walk, with or without aids, increased to 17 (p = 0.0001). We have experienced no evidence of epiphyseal damage after the procedure, and complication rates requiring rod exchange have been low (7%). PMID- 9853493 TI - A new method of stabilising the elevated laminae in open-door laminoplasty using an anchor system. AB - We treated 12 patients with multilevel stenosis of the cervical canal after spondylosis or ossification of the posterior longitudinal ligament by an expansive open-door laminoplasty, stabilised by using an anchor system. The preoperative sagittal diameter of the canal was 9.8 mm(+/-2.2) which was increased to 16.1 mm(+/-2.9) after surgery. The mean expansion ratio of the canal was 64% (42 to 100). The anchoring systems did not fail during the follow-up period (mean 29.5 months), and the decompression was maintained. The use of anchor systems to stabilise the posterior elements after laminoplasty is a simple and effective technique for maintaining the increased sagittal diameter of the canal. PMID- 9853494 TI - Spinal motion in lumbar degenerative disc disease. AB - We have measured the dynamic movement of the lumbar spine in 57 patients with degenerative lumbar disc disease. Each completed a questionnaire which recorded pain and subjective signs and symptoms. From plain lateral radiographs, the subjects were graded using the criteria of Kellgren and Lawrence and those of Lane et al, which are both based on the severity of degenerative changes. Measurements of the height of the disc space and the vertebral height were obtained and expressed as a ratio. We found no relationship between the characteristics of spinal movement and the overall grading of degenerative disc disease with either system. Both were influenced (p < 0.01) by age, walking distance, severity of symptoms, drug intake and frequency of pain. The present systems for grading degenerative disc disease from plain lateral radiographs have limited application. PMID- 9853495 TI - Dynamic external fixation for injuries of the proximal interphalangeal joint. AB - Pain, stiffness, instability and degenerative arthritis are common sequelae of complex fracture-dislocations of the proximal interphalangeal (PIP) joint. Operations were carried out to obtain stability, followed by application of a dynamic external fixator in 20 patients with a mean age of 29 years. This provided stability and distraction, and allowed controlled passive movement. Most (70%) of the patients had a chronic lesion and the mean time from injury to surgery was 215 days (3 to 1953). The final mean range of movement was 12 to 86 degrees. Complications included redislocation and septic arthritis, which affected the outcome. Four pin-track infections and two breakages of the hinge did not influence the result. The PIP Compass hinge is a useful adjunct to surgical reconstruction of the injured PIP joint. PMID- 9853496 TI - The use of external fixators for finger injuries: pin placement and tethering of the extensor hood. AB - External fixation is useful for the treatment of selected injuries to the hand. Some authors have suggested that external fixation of a phalanx may tether the extensor hood, thereby hindering active movements and predisposing to permanent adhesions. There is no consensus as to the best site for placement of the pin to minimise these problems. This study was performed on cadaver specimens to investigate the influence of the pin site on the range of simulated active movement of the interphalangeal joint. The dorsal midline position produces least interference with the extensor mechanism; radial and ulnar to this, interdigitating oblique fibres prevent a clean longitudinal split in the direction of gliding thus limiting movement of the extensor hood. At the proximal phalanx, positioning of the pin just off the midline avoids the thickening of the proximal median hood, whereas at the middle phalanx, a true midline position utilises the bare area at its base. PMID- 9853497 TI - Prognostic factors for major limb re-implantation at both immediate and long-term follow-up. AB - We performed a prospective study in 186 patients with a minimum follow-up of two years in order to attempt to predict successful major re-implantation of the upper limb. There were 137 men and 49 women with 24 amputations of the palm, 75 of the wrist, 50 of the forearm, 9 disarticulations through the elbow, and 28 amputations through the upper arm. The degree of injury to the amputated segment and the stump were good predictors of the rate of success and the final outcome. Adequate preservation, contraction of the muscle in the amputated part after stimulation, the level of injury and a smoking habit were fair indicators, but the serum potassium concentration in the amputated segment was the best objective predictor. When it is higher than 6.5 mmol/l 30 minutes after re-perfusion, re implantation should be avoided. A high systemic venous serum potassium concentration was also found before clinical signs of the re-perfusion syndrome were seen. PMID- 9853498 TI - Management of recurrent, complex instability of the elbow with a hinged external fixator. AB - We have treated 16 patients with recurrent complex elbow instability using a hinged external fixator. All patients had instability, dislocation or subluxation of the ulnohumeral joint. The injuries were open in eight patients and were associated with 20 other fractures and five peripheral nerve injuries. Two patients had received initial treatment from us; 14 had previously had a mean of 2.1 unsuccessful surgical procedures (1 to 6). The fixator was applied at a mean of 4.8 weeks (0 to 9) after the injury and remained on the elbow for a mean of 8.5 weeks (6 to 11). After treatment we found the mean range of flexion-extension to be 105 degrees (65 to 140). At a final follow-up of 23 months (14 to 40), the mean Morrey score was 84 (49 to 96): this translated into one poor, three fair, ten good and two excellent results. Complications included one fractured humeral pin, one temporary palsy of the radial nerve, one recurrent instability, one wound infection, one severe pin-track infection and one patient with reflex sympathetic dystrophy. Although technically demanding, the use of the fixator is an important advance in the management of recurrent complex elbow instability after failure of conventional treatment. PMID- 9853499 TI - A primary intrapelvic hydatid cyst presenting with foot drop and a gluteal swelling: a case report. AB - A primary hydatid cyst in the pelvis is rare, and usually presents with pressure symptoms affecting the adjacent abdominal organs. We describe such a cyst which protruded through the sciatic notch and presented as a gluteal swelling with a foot drop due to compression of the lumbosacral nerve roots. Surgical excision and postoperative treatment with albendazole for six weeks were effective in controlling the disease and preventing recurrence. PMID- 9853500 TI - The treatment of congenital brachymetatarsia by one-stage lengthening. AB - We performed one-stage lengthening using intercalary autogenous bone graft in 34 metatarsals and seven proximal phalanges in 21 patients with congenitally short metatarsals. At operation, in order to decrease the tension in the surrounding soft tissues, we gradually distracted the osteotomies of the affected bones for 20 to 30 minutes. The patients, all women, were followed up for a mean period of 2.1 years (1 to 6.5). The average gain in length for the 34 metatarsal procedures was 14 mm (6 to 21), equivalent to an increase of 32% (11 to 51), and for the seven proximal phalangeal lengthenings 8 mm (5 to 11), an increase of 54% (47 to 65). There was no evidence of neurovascular impairment. The technique of gradual distraction during operation is simple and effective. It overcomes the disadvantages of one-stage lengthening such as a small gain in length and neurovascular damage. PMID- 9853501 TI - The use of antibiotic-impregnated cement in infected reconstructions after resection for bone tumours. AB - We have studied 35 patients with infected reconstructions after segmental resection for bone tumours. Two different regimes of treatment were employed both of which entailed debridement and the use of cement impregnated with antibiotic. In 25 patients gentamicin-PMMA beads were used and in ten an antibiotic impregnated cement spacer. Better results were achieved with the second procedure in terms of cases healed, the number of operations, time of healing, time of recovery and the functional score. The use of vancomycin in the cement spacer gave better local control. PMID- 9853502 TI - Genotype phenotype correlation in achondroplasia and hypochondroplasia. AB - Recent studies of the fibroblast growth factor receptor 3 (FGFR3) gene have established that achondroplasia and hypochondroplasia are allelic disorders of different mutations. To determine whether the genotype could be distinguished on the basis of the phenotype, we analysed height, arm span, and skeletal radiographs from 23 patients with achondroplasia and the G380R mutation of FGFR3 and eight with hypochondroplasia and the N540K mutation. Both conditions share the classical pathological features of micromelic short stature, reduced or unchanged interpedicular distances in the lumbar spine, disproportionately long fibulae, and squared and shortened pelvic ilia. These were significantly more severe in the G380R patients than in the N540K patients. Our findings have shown a firm statistical correlation between the genotype and the phenotype, although there were a few exceptional cases in which there was phenotypic overlap between the two conditions. PMID- 9853503 TI - Venous haemodynamics after total knee arthroplasty: evaluation of active dorsal to plantar flexion and several mechanical compression devices. AB - We performed a crossover study to evaluate the haemodynamic effect of active dorsal to plantar flexion and seven pneumatic compression devices in ten patients who had a total knee arthroplasty. Using the Acuson 128XP/10 duplex ultrasound unit with a 5MHz linear array probe, we assessed the augmentation of peak venous velocity and venous volume above and below the junction of the greater saphenous and common femoral veins in order to study both the deep and superficial venous systems. The pneumatic compression devices evaluated included two foot pumps (A-V Impulse System and PlexiPulse Foot), a foot-calf pump (PlexiPulse Foot-Calf), a calf pump (VenaFlow System) and three calf-thigh pumps (SCD System, Flowtron DVT and Jobst Athrombic Pump). The devices differed in a number of ways, including the length and location of the sleeve and bladder, the frequency and duration of activation, the rate of pressure rise, and the maximum pressure achieved. A randomisation table was used to determine the order of the test conditions for each patient. The enhancement of peak venous velocity occurred primarily in the deep venous system below the level of the saphenofemoral junction. The increases in peak venous velocity were as follows: active dorsal to plantar flexion 175%; foot pumps, A-V Impulse System 29% and PlexiPulse 65%; foot-calf pump, PlexiPulse, 221%; calf pump, VenaFlow, 302% and calf-thigh pumps, Flowtron DVT 87%, SCD System 116% and Jobst Athrombic Pump 263%. All the devices augmented venous volume, the greatest effect being seen with those incorporating calf compression. The increases in ml/min were found in the deep venous system as follows: foot pumps, A-V Impulse System 9.6 and PlexiPulse Foot 16.7; foot-calf pump, PlexiPulse, 38.1; calf pump, VenaFlow, 26.2; calf-thigh pumps, Flowtron DVT 61.5, SCD System 34.7 and Jobst Athrombic Pump 82.3. Active dorsal to plantar flexion generated 8.5 ml for a single calf contraction. PMID- 9853504 TI - Observations on the direction of wear in Charnley sockets retrieved at revision. AB - The direction of wear in the acetabular socket has implications for the amount of debris that is generated during movement, for the magnitude of eccentric loading and for the incidence of impingement of the neck. We observed the direction of penetration with respect to a global co-ordinate system in 84 acetabular components retrieved at reoperation. The mean direction of wear relative to the open face of the sockets was found to be 37 degrees with a range from 0 degrees to 87 degrees. For those values determined using the inclination of the socket on the prerevision radiograph, the mean direction of penetration in the coronal plane had a lateral, rather than a medial, component. The mean angle was 84 degrees (SD 17 degrees) with respect to the horizontal. The angle of penetration was found to correlate significantly with the depth, in that the lateral component became larger as the wear progressed. There was also a significant correlation between the rate of penetration and the direction of wear. Despite the theoretical advantage of penetration in the superolateral direction, i.e., along the margin of the socket, in reducing the probability of impingement of the neck, no significant correlation was seen between the angle of penetration and the period of use in vivo. This may suggest that impingement of the femoral neck on the rim of the socket may not be the dominant factor in loosening of the socket but can still be important in a few cases. PMID- 9853505 TI - Spinal canal capacity in simulated displacements of the atlantoaxial segment: a skeletal study. AB - In normal, physiological circumstances there is ample room in the spinal canal to accommodate the spinal cord. Our study aimed to identify the degree of compromise of the spinal canal which could be anticipated in various atlantoaxial pathological states. We examined paired atlas and axis vertebrae using high definition radiography and simultaneous photography in both normal and simulated pathological orientations in order to measure the resultant dimension of the spinal canal and its percentage occlusion. At the extreme of physiological axial rotation (47 degrees) the spinal canal is reduced to 61% of its cross-sectional area in neutral rotation. The spinal cord is thus safe from compromise. Atlantoaxial subluxation of up to 9 mm reduces the area of the spinal canal, in neutral rotation, to 60% with no cord compromise. Any rotation is, however, likely to cause cord compression. The mechanism of fixation in atlantoaxial rotatory subluxation could be explained by bony interlocking of the facet joint, reproducible in dry bones. PMID- 9853506 TI - Academic orthopaedics: a discipline at risk. PMID- 9853507 TI - Academic orthopaedics: a discipline at risk. PMID- 9853508 TI - Academic orthopaedics: a discipline at risk. PMID- 9853509 TI - Academic orthopaedics: a discipline at risk. PMID- 9853511 TI - Academic orthopaedics: a discipline at risk. PMID- 9853510 TI - Academic orthopaedics: a discipline at risk. PMID- 9853512 TI - The search for the "best" cytokine to induce antitumor immunity. PMID- 9853513 TI - Insertion of two independent enhancers in the long terminal repeat of a self inactivating vector results in high-titer retroviral vectors with tissue-specific expression. AB - The use of retroviral vectors (RVs) derived from the murine oncoretroviruses for gene therapy is associated with the risk of malignant transformation of infected cells and ectopic expression of the proteins of interest. Targeting retroviral vectors to specific tissues would increase their safety and clinical applicability. To explore the potential of targeting vector expression to skeletal muscle, the murine leukemia virus broad transcriptional tropism was modified by substituting the viral promoter and/or enhancer with a transcriptional cassette containing the human T cell leukemia virus type I Tax responsive element and the minimal muscle creatine kinase enhancer and promoter. The resulting retroviral vectors could be transcriptionally trans-activated by tax. In the absence of Tax, however, the viruses showed muscle-specific expression. Trans-complementing packaging and indicator cells stably expressing Tax were used to isolate high-titer producer cell clones (10(6) CFU/ml). In vitro, the levels of expression of these RVs in Tax-expressing fibroblasts were 10,000-fold higher than in normal fibroblasts and 1000-fold higher in C2C12 myotubes than in C2C12 myoblasts. Expression of the vectors and the endogenous muscle creatine kinase gene was similarly dependent on the maturity of the muscle cultures. One vector with modified LTRs was also tested in vivo in regenerating muscle and showed a delayed pattern of expression in myofibers compared with the vector containing the wild-type LTRs. These vectors can be easily modified to contain different tissue-specific enhancer and promoter elements and the availability of complementing packaging and indicator cells expressing Tax should allow their application in a variety of gene therapy settings. PMID- 9853514 TI - Amphotropic retroviral vectors displaying hepatocyte growth factor-envelope fusion proteins improve transduction efficiency of primary hepatocytes. AB - The development of retroviral vectors with cell-specific targeting capabilities will be an important step toward successful in vitro gene therapy. This article describes the generation of a retroviral vector with enhanced binding abilities for cells bearing the c-Met receptor: the Madin-Darby canine kidney (MDCK) cell line and primary hepatocytes. The human hepatocyte growth factor (HGF) was displayed on murine amphotropic retroviral vectors by fusion to the viral transmembrane envelope glycoprotein (TM). The resulting chimeric envelope HGF-TM was expressed in an amphotropic packaging cell line producing viral particles that display both HGF-TM and the wild-type envelope. These modified viral particles had a titer equivalent to that of unmodified particles. Modified particles infected MDCK cells more efficiently than did unmodified amphotropic retrovirus. Adding anti-HGF antibodies to the viral vector particle supernatant prior to infection confirmed that the increased infection was mediated by the HGF moiety. The chimeric viruses also infected primary mouse and nonhuman primate fetal hepatocytes more effectively. Furthermore, these cells could be induced to proliferate by the modified HGF-TM viruses. Since exogenous HGF is primarily taken up by the liver, these results may have implications for retroviral vector design for liver-directed human gene therapy. PMID- 9853515 TI - Canarypox virus-mediated interleukin 12 gene transfer into murine mammary adenocarcinoma induces tumor suppression and long-term antitumoral immunity. AB - The antitumoral activity of recombinant canarypox virus vectors (ALVAC) expressing murine interleukin 12 (IL-12) was evaluated in the syngeneic, nonimmunogenic murine mammary adenocarcinoma model (TS/A). Seven-day preestablished subcutaneous tumors (5- to 6-mm mean diameters) were injected on days 7, 10, 14, 17, 21, and 24 with the vector ALVAC-IL12 at 2.5 x 10(5) TCID50 (50% tissue culture infective dose). Total tumor regression occurred in 40 to 50% of the treated mice. Furthermore, 100% of the cured mice were protected against a contralateral subsequent challenge with the TS/A parental cells on day 28. The ALVAC-IL12 treatment is not effective in nude mice, suggesting the critical role of T cells. CD4 and CD8 T cells infiltrated the tumors treated with ALVAC-IL12 in the BALB/c model. Furthermore, in vivo depletion of CD4+ T cells totally abrogated the induction of the long-term antitumoral immune response by ALVAC IL12. Interestingly, some tumor growth inhibition was also observed with ALVAC betaGal treatment and a vaccinal effect was found in 33% of the treated animals, suggesting an adjuvant effect of the vector itself. Other ALVAC vectors expressing murine cytokines (IL-2, GM-CSF, IFN-gamma) were evaluated in the same model. Major antitumoral activity was observed with ALVAC-GM-CSF. However, a combination of ALVAC-GM-CSF and ALVAC-IL12 had no synergistic effect. These results suggest that in vivo gene transfer with canarypox virus expressing IL-12 may provide an effective and safe strategy for the treatment of human cancers. PMID- 9853516 TI - Lipofectamine and related cationic lipids strongly improve adenoviral infection efficiency of primitive human hematopoietic cells. AB - Adenoviral vectors have the potential to infect a large number of cell types including quiescent cells. Their use in hematopoietic cells is limited by the episomal form of their DNA, leading to transgene loss in the progeny cells. However, the use of this vector may be interesting for short-term in vitro modifications of primitive human hematopoietic cells. Therefore, we have investigated the ability of adenovirus to transduce cord blood CD34+ cells. Several promoters were tested using the lacZ reporter gene. The PGK and CMV promoters induced transgene expression in 18-25% of the cells, whereas the HTLV-I and especially the RSV promoter were almost inactive. To improve infection efficiency, adenovirus was complexed with cationic lipids. Lipofectamine, Cellfectin, and RPR120535b, but not Lipofectin, Lipofectace, or DOTAP, markedly improved transgene expression in CD34+ cells (from 19 to 35%). Lipofectamine strongly enhanced infection efficiency of the poorly infectable primitive CD34+CD38low cells (from 11 to 28%) whereas the more mature CD34+CD38+ cells were only slightly affected (from 24 to 31%). Lipofectamine tripled the infection of CFU-GMs and LTC-ICs derived from the CD34+CD38low cell fraction (from 4 to 12% and from 5 to 16%, respectively) and doubled that of BFU-Es (from 13 to 26%). We conclude that cationic lipids can markedly increase the efficiency of adenovirus mediated gene transfer into primitive hematopoietic cells. PMID- 9853517 TI - Generation of fiber-mutant recombinant adenoviruses for gene therapy of malignant glioma. AB - Recombinant adenovirus (Adv)-mediated gene transduction is a powerful technology for cancer gene therapy. In this article, we report the generation of a fiber mutant Adv vector, using the Adv genomic DNA-terminal protein complex (DNA-TPC) cotransfection method. First, a fiber-mutant construct in a plasmid carrying the right-side two-thirds of the human adenovirus type 5 (Ad5) genome (pTR) was cotransfected with Ad5 DNA-TPC, yielding the recombinant Adv with the desired fiber mutation. The DNA-TPC from the mutant Adv was then utilized to produce a second-step recombinant Adv with an expression cassette in the place of E1. By this procedure, we generated a fiber mutant, F/K20, that has a linker and a stretch of 20 lysine residues added at the C terminus of the fiber. By using Adv carrying a reporter lacZ gene (AxCAZ2) with either F/K20 or wild-type fiber (F/wt), we examined the transduction efficiency of F/K20-Adv. No significant difference in the transduction efficiency between F/K20 and F/wt-Adv was observed for a human fibroblast line, WI-38, or various tumor cell lines, including melanoma, prostate, esophageal, and pancreatic cancer lines. In clear contrast, F/K20-Adv showed a remarkably enhanced efficiency in genetic transduction of human glioma cells. In all four human glioma lines tested, the multiplicities of infection (MOIs) for transduction of 50% of the population (ED50) were decreased with F/K20-Adv compared with F/wt-Adv: 7-fold for T98G, 14-fold for U251, 9-fold for U373, and 42-fold for U87 cells. Therefore, we attempted to apply F/K20-Adv for gene therapy of malignant glioma. Glioma cells infected with F/K20-Adv carrying genes for interleukin 2 or interleukin 12 produced a high level of each cytokine at a much lower MOI than did cells infected with F/wt-Adv. Infection with F/K20-Adv carrying the wild-type p53 tumor suppressor gene resulted in an enhanced level of p53 protein expression and an increased incidence of F/K20-Adv in transduction efficiency for malignant glioma, providing promising tools for gene therapy. PMID- 9853518 TI - Helper-free foamy virus vectors. AB - Retroviral vectors based on human foamy virus (HFV) have been developed and show promise as gene therapy vehicles. Here we describe a method for the production of HFV vector stocks free of detectable helper virus. The helper and vector plasmid constructs used both lack the HFV bel genes, so recombination between these constructs cannot create a wild-type virus. A fusion promoter that combines portions of the cytomegalovirus (CMV) immediate-early and HFV long terminal repeat (LTR) promoters was used to drive expression of both the helper and vector constructs. The CMV-LTR fusion promoter allows for HFV vector production in the absence of the Bel-1 trans-activator protein, which would otherwise be necessary for efficient transcription from the HFV LTR. Vector stocks containing either neomycin phosphotransferase or alkaline phosphatase reporter genes were produced by transient transfection at titers greater than 10(5) transducing units/ml. G418 resistant BHK-21 cells obtained by transduction with neo vectors contained randomly integrated HFV vector proviruses without detectable deletions or rearrangements. The vector stocks generated were free of replication-competent retrovirus (RCR), as determined by assays for LTR trans-activation and a marker rescue assay developed here for the detection of Bel-independent RCR. PMID- 9853519 TI - Behavioral recovery in 6-hydroxydopamine-lesioned rats by cotransduction of striatum with tyrosine hydroxylase and aromatic L-amino acid decarboxylase genes using two separate adeno-associated virus vectors. AB - Parkinson's disease (PD) is characterized by the progressive loss of the dopaminergic neurons in the substantia nigra and a severe decrease in dopamine in the striatum. A promising approach to the gene therapy of PD is intrastriatal expression of enzymes in the biosynthetic pathway for dopamine. Tyrosine hydroxylase (TH) catalyzes the synthesis of L-dopa, which must be converted to dopamine by aromatic L-amino acid decarboxylase (AADC). Since the endogenous AADC activity in the striatum is considered to be low, coexpression of both TH and AADC in the same striatal cells would increase the dopamine production and thereby augment the therapeutic effects. In the present study, the TH gene and also the AADC gene were simultaneously transduced into rat striatal cells, using two separate adeno-associated virus (AAV) vectors, AAV-TH and AAV-AADC. Immunostaining showed that TH and AADC were coexpressed efficiently in the same striatal cells in vitro and in vivo. Moreover, cotransduction with these two AAV vectors resulted in more effective dopamine production and more remarkable behavioral recovery in 6-hydroxydopamine (6-OHDA)-lesioned rats, compared with rats receiving AAV-TH alone (p < 0.01). These findings suggest an alternative strategy for gene therapy of PD and indicate that the simultaneous transduction with two AAV vectors can extend their utility for potential gene therapy applications. PMID- 9853520 TI - Coexpression of cytidine deaminase and mutant dihydrofolate reductase by a bicistronic retroviral vector confers resistance to cytosine arabinoside and methotrexate. AB - The transfer of a drug resistance gene into hematopoietic cells is an approach being investigated to overcome the problem of myelosuppression produced by anticancer drugs. Chemotherapeutic agents are often given in combination in order to increase their effectiveness. Consequently, there is an advantage in designing vectors for gene transfer that are capable of expressing two drug resistance genes. We have constructed a bicistronic retroviral vector, MFG-DHFR-IRES/CD, which contains the mutated human dihydrofolate reductase (DHFR) cDNA with a phenylalanine-to-serine substitution at codon 31 (F31S) and the human cytidine deaminase (CD) cDNA. Murine fibroblast and hematopoietic cells were transduced with this vector and evaluated for their resistance to methotrexate (MTX) and cytosine arabinoside (ARA-C). The transduced fibroblast cells showed high levels of resistance to MTX and to ARA-C as determined by a clonogenic assay. Using enzymatic assays, we observed a coordinate increase in resistance to MTX and DHFR enzyme activity following an ARA-C selection. In addition, MTX selection produced an increase in CD enzyme activity and ARA-C resistance. Murine hematopoietic cells transduced with the bicistronic vector also showed drug resistance to both MTX and ARA-C. Interestingly, the double-gene construct conferred an equivalent level of drug resistance compared with single-gene vectors bearing only CD or DHFR genes in the hematopoietic cells. These results demonstrate the potential of the MFG-DHFR-IRES/CD vector to confer drug resistance to both MTX and ARA-C and may have future application in chemoprotection of normal hematopoietic cells in patients with cancer. PMID- 9853521 TI - Enhancer and promoter chimeras in plasmids designed for intramuscular injection: a comparative in vivo and in vitro study. AB - Enhancers and promoters from various muscle-specific genes were substituted for or combined with the enhancer/promoter of the human cytomegalovirus (CMV) IE gene in a luciferase reporter gene plasmid in an effort to identify new promoter chimeras with increased expression activity after direct intramuscular injection. The regulatory sequence substitutions or additions varied in content, location, and orientation relative to the CMV regulatory sequences. The expression activities of the derivative and parent plasmids were compared quantitatively in vivo using a standard mouse intramuscular injection assay, and in vitro by transfection of differentiated C2C12 mouse myoblasts and BHK hamster kidney cells, to test whether cultured cell transfection could substitute for at least some animal experimentation. In vivo, 1 of 19 of the enhancer/promoter chimeras increased expression levels. In vitro, some chimeras showed significant expression augmentation in C2C12 cells, but not in BHK cells. We conclude that because of differences in plasmid expression profiles, these cell culture systems cannot readily substitute for in vivo testing of new plasmid constructs. PMID- 9853522 TI - Attenuation of skeletal muscle wasting with recombinant human growth hormone secreted from a tissue-engineered bioartificial muscle. AB - Skeletal muscle wasting is a significant problem in elderly and debilitated patients. Growth hormone (GH) is an anabolic growth factor for skeletal muscle but is difficult to deliver in a therapeutic manner by injection owing to its in vivo instability. A novel method is presented for the sustained secretion of recombinant human GH (rhGH) from genetically modified skeletal muscle implants, which reduces host muscle wasting. Proliferating murine C2C12 skeletal myoblasts stably transduced with the rhGH gene were tissue engineered in vitro into bioartificial muscles (C2-BAMs) containing organized postmitotic myofibers secreting 3-5 microg of rhGH/day in vitro. When implanted subcutaneously into syngeneic mice, C2-BAMs delivered a sustained physiologic dose of 2.5 to 11.3 ng of rhGH per milliliter of serum. rhGH synthesized and secreted by the myofibers was in the 22-kDa monomeric form and was biologically active, based on downregulation of a GH-sensitive protein synthesized in the liver. Skeletal muscle disuse atrophy was induced in mice by hindlimb unloading, causing the fast plantaris and slow soleus muscles to atrophy by 21 to 35% ( < 0.02). This atrophy was significantly attenuated 41 to 55% (p < 0.02) in animals that received C2-BAM implants, but not in animals receiving daily injections of purified rhGH (1 mg/kg/day). These data support the concept that delivery of rhGH from BAMs may be efficacious in treating muscle-wasting disorders. PMID- 9853523 TI - Targeted delivery of DNA encoding cytotoxic proteins through high-affinity fibroblast growth factor receptors. AB - Nonviral DNA delivery strategies for gene therapy have generally been limited by a lack of specificity and efficacy. However, ligand-mediated endocytosis can specifically deliver DNA in vitro to cells bearing the appropriate cognate receptors. Similarly, in order to circumvent problems related to efficacy, DNA must encode proteins with high intrinsic activities. We show here that the ligand basic fibroblast growth factor (FGF2) can target FGF receptor-bearing cells with DNA encoding therapeutic proteins. Delivery of genes encoding saporin, a highly potent ribosomal inactivating protein, or the conditionally cytotoxic herpes simplex virus thymidine kinase, a protein that can kill cells by activating the prodrug ganciclovir, is demonstrated. The saporin gene was codon optimized for mammalian expression and demonstrated to express functional protein in a cell free assay. FGF2-mediated delivery of saporin DNA or thymidine kinase DNA followed by ganciclovir treatment resulted in a 60 and 75% decrease in cell number, respectively. Specificity of gene delivery was demonstrated in competition assays with free FGF2 or with recombinant soluble FGF receptor. Alternatively, when histone H1, a ligand that binds to cell surface heparan sulfate proteoglycans ("low-affinity" FGF receptors), was used to deliver DNA encoding thymidine kinase, no ganciclovir sensitivity was observed. These findings establish the feasibility of using ligands such as FGF2 to specifically deliver genes encoding molecular chemotherapeutic agents to cells. PMID- 9853524 TI - Efficient construction of a recombinant adenovirus vector by an improved in vitro ligation method. AB - An efficient method for constructing a recombinant adenovirus (Ad) vector, based on an in vitro ligation, has been developed. To insert the foreign gene into an adenoviral DNA, we introduced three unique restriction sites, I-CeuI, SwaI, and PI-SceI, into the E1 deletion site of the vector plasmid, which contains a complete E1, E3-deleted adenovirus type 5 genome. I-CeuI and PI-SceI are intron encoded endonucleases with a sequence specificity of at least 9-10 and 11 bp, respectively. A shuttle plasmid, pHM3, containing multiple cloning sites between the I-CeuI and PI-SceI sites, was constructed. After the gene of interest was inserted into this shuttle plasmid, the plasmid for E1-deleted adenovirus vector could be easily prepared by in vitro ligation using the I-CeuI and PI-SceI sites. SwaI digestion of the ligation products prevented the production of a plasmid containing a parental adenovirus genome (null vector). After transformation into E. coli, more than 90% of the transformants had the correct insert. To make the vector, a PacI-digested, linearized plasmid was transfected into 293 cells, resulting in a homogeneous population of recombinant virus. The large number and strategic location of the unique restriction sites will not only increase the rapidity of production of new first-generation vectors for gene transfer but will allow for rapid further improvements in the vector DNA backbone. PMID- 9853525 TI - A phase I/II dose-escalation study of herpes simplex virus type 1 thymidine kinase "suicide" gene therapy for metastatic melanoma. Study Group on Gene Therapy of Metastatic Melanoma. AB - We performed a dose-escalating phase I/II study of retrovirus-mediated herpes simplex virus type 1 thymidine kinase (HSV-1-TK) suicide gene therapy for metastatic melanoma. HSV-1 TK expression, which specifically sensitizes transduced and bystander cancer cells to ganciclovir (GCV) toxicity, was mediated by one (four patients, first dose step) to three (four patients, second dose step) injections of "M11" retrovirus vector-producing cells in melanoma cutaneous nodules. After a 7-day period allowed for cancer cell transduction, GCV was administered for 14 days. Safety was assessed by clinical and laboratory evaluations, and efficacy was assessed by tumor measurements and histology. M11 doses ranged from 76 to 1247 x 10(6) cells. Treatment-related adverse events were mild and transient, limited to inflammatory skin reactions at injection and fever on repeated injections. Plasma GCV was in the active range (>0.2 microg/ml); transgene was detected by polymerase chain reaction in three of six patients; treated tumor size was moderately affected under GCV as compared with untreated tumors, although 2 weeks after GCV administration important (>50%) treated-tumor necrosis was evidenced on histology in three of eight patients. All patients showed disease progression on long-term follow-up. Thus, M11-mediated HSV-1 TK gene therapy was well tolerated over a wide dose range. The limited tumor response is likely to be related to poor gene transfer efficiency. However, necrosis following GCV administration in transduced tumors indicates a potential for treatment efficacy. PMID- 9853526 TI - A phase I/II study of herpes simplex virus type 1 thymidine kinase "suicide" gene therapy for recurrent glioblastoma. Study Group on Gene Therapy for Glioblastoma. AB - Despite extensive surgery for glioblastoma, residual tumor cells always lead to relapse. Gene therapy based on retrovirus-mediated gene transfer of herpes simplex virus type 1 thymidine kinase (HSV-1 TK), which specifically sensitizes dividing cells to ganciclovir (GCV) toxicity, may help eradicate such cells. During glioblastoma surgery, HSV-1 TK retroviral vector-producing cells (M11) were injected into the surgical cavity margins after tumor debulking. After a 7 day transduction period, GCV was administered for 14 days. Safety was assessed by clinical and laboratory evaluations, and efficacy was assessed by MRI-based relapse-free survival at month 4 and by overall survival. Twelve patients with recurrent glioblastoma were treated without serious adverse events related to M11 cell administration or GCV. Quality of life was not negatively influenced by this treatment. Overall median survival was 206 days, with 25% of the patients surviving longer than 12 months. At 4 months after treatment, 4 of 12 patients had no recurrence; their median overall survival was 528 days, compared with 194 days for patients with recurrence (p=0.03 by the log rank test). One patient is still free of detectable recurrence, steroid free and independent, 2.8 years after treatment. Thus, brain injections of M11 retroviral vector-producing cells for glioblastoma HSV-1 TK gene therapy were well tolerated and associated with significant therapeutic responses. These results warrant further development of this therapeutic strategy in brain tumor, including recurrent glioblastoma. PMID- 9853527 TI - Central nervous system delivery of interleukin 4 by a nonreplicative herpes simplex type 1 viral vector ameliorates autoimmune demyelination. AB - Multiple sclerosis (MS) is a T cell-mediated organ-specific inflammatory disease leading to central nervous system (CNS) demyelination. On the basis of results obtained in experimental autoimmune encephalomyelitis (EAE) models, MS treatment by administration of antiinflammatory cytokines such as interleukin 4 (IL-4) is promising but is hampered by the limited access of the cytokines to the CNS and by the pleiotropic effects of systemically administered cytokines. We established a cytokine delivery system within the CNS using non-replicative herpes simplex type 1 (HSV-1) viral vectors engineered with cytokine genes. These vectors injected into the cisterna magna (i.c.) of mice diffuse in all ventricular and subarachnoid spaces and infect with high efficiency the ependymal and leptomeningeal cell layers surrounding these areas, without obvious toxic effects. Heterologous genes contained in the vectors are efficiently transcribed in infected ependymal cells, leading to the production of high amounts of the coded proteins. For example, 4.5 ng of interferon gamma (IFN-gamma) per milliliter is secreted into the cerebrospinal fluid (CSF) up to day 28 postinjection (p.i.) and reaches the CNS parenchyma in bioactive form, as demonstrated by upregulation of MHC class I expression on CNS-resident cells. We then exploited the therapeutic potential of the vectors in EAE mice. An HSV-1 derived vector containing the IL-4 gene was injected i.c. in Biozzi AB/H mice at the time of EAE induction. We found the following in treated mice: (1) delayed EAE onset, (2) a significant decrease in clinical score, (3) a significant decrease in perivascular inflammatory infiltrates and in the number of macrophages infiltrating the CNS parenchyma and the submeningeal spaces, and (4) a reduction in demyelinated areas and axonal loss. Peripheral T cells from IL-4 treated mice were not affected either in their antigen-specific proliferative response or in cytokine secretion pattern. Our results indicate that CNS cytokine delivery with HSV-1 vectors is feasible and might represent an approach for the treatment of demyelinating diseases. Advantages of this approach over systemic cytokine administration are the high cytokine level reached in the CNS, the absence of effects on the peripheral immune system, and the long-lasting cytokine production in the CNS after a single vector administration. PMID- 9853528 TI - RNA levels of human retrovirus receptors Pit1 and Pit2 do not correlate with infectibility by three retroviral vector pseudotypes. AB - The gibbon ape leukemia virus (GaLV) and the amphotropic murine leukemia virus (A MuLV) infect human cells via specific receptors, Pit1 and Pit2, respectively. mRNA levels of these receptors were determined by Northern analysis and for Pit2 in addition by quantitative RT-PCR. Pit1 and Pit2 were expressed in different amounts in human tissues and cell lines; Pit1-specific mRNA was generally more abundant than Pit2 mRNA. No correlation was found between Pit1 and Pit2 RNA levels and infectibility by GaLV and A-MuLV pseudotyped vectors, respectively. GaLV and A-MuLV revealed a partial reciprocal interference. MuLV-10A1 can utilize both Pit1 and Pit2 for entry into cells but could not infect any of the 14 human cell lines more efficiently than A-MuLV or GaLV. Interference assays suggested that MuLV-10A1 has a higher affinity for and infected most cells predominantly by Pit2. However, at least in one cell line it used Pit1 more efficiently for entry. We conclude that (1) Pit1 and Pit2 mRNA levels in human cells are not indicative of the infectibility by GaLV and A-MuLV pseudotypes, respectively; (2) A-MuLV can infect target cells as efficiently as can GaLV, although Pit2 RNA is less abundant than Pit1 RNA; (3) factor(s) in addition to the presence of Pit1 and Pit2 are involved in retroviral infection; and (4) MuLV-10A1 pseudotype does not infect human cells more efficiently than do A-MuLV and GaLV pseudotypes. PMID- 9853529 TI - Retroviral transfer of the glucocerebrosidase gene into CD34+ cells from patients with Gaucher disease: in vivo detection of transduced cells without myeloablation. AB - Retroviral gene transfer of the glucocerebrosidase gene to hematopoietic progenitor and stem cells has shown promising results in animal models and corrected the enzyme deficiency in cells from Gaucher patients in vitro. Therefore, a clinical protocol was initiated to explore the safety and feasibility of retroviral transduction of peripheral blood (PB) or bone marrow (BM) CD34+ cells with the G1Gc vector. This vector uses the viral LTR promoter to express the human glucocerebrosidase cDNA. Three adult patients have been entered with follow-up of 6-15 months. Target cells were G-CSF-mobilized and CD34 enriched PB cells or CD34-enriched steady state BM cells, and were transduced ex vivo for 72 hr. Patient 1 had PB cells transduced in the presence of autologous stromal marrow cells. Patient 2 had PB cells transduced in the presence of autologous stroma, IL-3, IL-6, and SCF. Patient 3 had BM cells transduced in the presence of autologous stroma, IL-3, IL-6, and SCF. At the end of transduction, the cells were collected and infused immediately without any preparative treatment of the patients. The transduction efficiency of the CD34+ cells at the end of transduction was approximately 1, 10, and 1 for patients 1, 2, and 3, respectively, as estimated by semiquantitative PCR on bulk samples and PCR analysis of individual hematopoietic colonies. Gene marking in vivo was demonstrated in patients 2 and 3. Patient 2 had vector-positive PB granulocytes and mononuclear bone marrow cells at 1 month postinfusion and positive PB mononuclear cells at 2 and 3 months postinfusion. Patient 3 had a positive BM sample at 1 month postinfusion but was negative thereafter. These results indicate that gene-marked cells can engraft and persist for at least 3 months postinfusion, even without myeloablation. However, the level of corrected cells (<0.02%) is too low to result in any clinical benefit, and glucocerebrosidase enzyme activity did not increase in any patient following infusion of transduced cells. Modifications of vector systems and transduction conditions, along with partial myeloablation to allow higher levels of engraftment, may be necessary to achieve beneficial levels of correction in patients with Gaucher disease. PMID- 9853530 TI - The treatment of malignant mesothelioma with a gene modified cancer cell line: a phase I study. AB - Malignant mesothelioma is a tumor of the pleura for which there is no satisfactory treatment. It is almost universally fatal, regardless of the stage of the tumor at the time of diagnosis. Current treatment modalities include surgery, chemotherapy, and radiation therapy, although in some series none of these modalities is superior to no treatment at all. Because of the dismal prognosis for patients with malignant mesothelioma, a new mode of treatment is desperately needed. A promising area of research into the treatment of various malignancies is gene therapy. Recent studies have demonstrated the utility of exposing tumor cells to cells transduced to express the Herpes simplex virus gene for thymidine kinase (HSV-tk). By virtue of their expression of HSV-tk, the transduced cells are rendered susceptible to the antiviral drug, ganciclovir (GCV). and nearby tumor cells are killed by a phenomenon termed the bystander effect. In this protocol we propose a Phase I trial to study the safety and determine the maximal tolerated dose of an HSV-tk-transduced ovarian cancer cell line (PA1-STK cells) infused into the pleural cavities of patients with malignant pleural mesothelioma, followed by systemic administration of ganciclovir. The hope is that administration of ganciclovir will result in killing of the HSV-tk transduced ovarian cancer cells as well as the nearby malignant mesothelioma cells. This is a standard dose-escalation protocol. PMID- 9853531 TI - Zinc deficiency in humans: a neglected problem. PMID- 9853532 TI - Chromium research from a distance: from 1959 to 1980. AB - More than 50 years of work have led to the recognition of trivalent chromium as an essential element. Shortly after its identification as an essential element in 1959, its interaction with insulin in vitro and in vivo was established, and the site of action identified as the insulin-sensitive cell membrane. Despite other early clinical successes with chromium supplementation, four major problems have influenced the rate of progress since then: 1) chromium analysis; 2) interaction of chromium with other dietary factors; 3) diagnosis of chromium status; and 4) other controversies, such as the carcinogenic potential of chromium (since disproved) and the lack of an effect on glucose tolerance even in chromium deficient organisms (now explained). These controversies have mostly dissipated as new knowledge integrated seemingly irreconcilable facts and opinions. It is now known that chromium may potentiate the action of insulin either by an effect on insulin dependent functions, or by maintaining these functions with less insulin, or by a combination of both. Despite much progress in the last 30 years, major challenges in chromium research remain, such as the development of practical methods for diagnosing chromium deficiency. Of several approaches for solving this problem, the most feasible might be to standardize the urinary chromium response following an insulinogenic challenge, such as an oral load of glucose or of glucose plus fructose (for maximal stimulation) with urine collection before and during the 2-hour test. PMID- 9853533 TI - Chromium, glucose intolerance and diabetes. AB - Within the last 5 years chromium (Cr) has been shown to play a role in glucose intolerance, Type 2 diabetes mellitus (Type 2 DM), and gestational diabetes. In addition, diabetes and the neuropathy of a patient on home parenteral nutrition were alleviated when supplemental Cr was added to total parenteral nutrition (TPN) solutions. In a study conducted in China that has been supported by studies in the United States, supplemental Cr as Cr picolinate improved the blood glucose, insulin, cholesterol, and hemoglobin A1C in people with Type 2 DM in a dose dependent manner. Follow-up studies of > 1 year have confirmed these studies. The requirement for Cr is related to the degree of glucose intolerance: 200 microg/day of supplemental Cr is adequate to improve glucose variables of those who are mildly glucose intolerant. However, people with more overt impairments in glucose tolerance and diabetes usually require more than 200 microg/day. Daily intake of 8 microg of Cr per kg body weight was also more effective than 4 microg/kg in women with gestational diabetes. The mechanism of action of Cr involves increased insulin binding, increased insulin receptor number, and increased insulin receptor phosphorylation. In summary, supplemental Cr has been shown to have beneficial effects without any documented side effects on people with varying degrees of glucose intolerance ranging from mild glucose intolerance to overt Type 2 DM. PMID- 9853534 TI - Zinc metabolism in patients with exocrine pancreatic insufficiency. AB - OBJECTIVE: Metabolism of zinc was studied by administering radioactive zinc (Zn65), and by performing metabolic balance studies in eight patients with exocrine pancreatic insufficiency and eighteen control subjects. METHODS: Retention of radioactive zinc was measured by total body counter, and its urinary and fecal excretion by gamma scintillation counter. Metabolic balance studies were carried out by measuring dietary zinc intake as well as fecal and urinary excretion of zinc by atomic absorption spectrophotometry in this group of patients. RESULTS: These studies revealed a 50% reduction in intestinal absorption of Zn65 in patients with exocrine pancreatic insufficiency as compared to alcoholic and non-alcoholic control subjects. In addition, there was a 2 to 4 fold increase (p<0.05) in urinary excretion of zinc in subjects with pancreatic disease. In pancreatic insufficiency, reduced zinc absorption and increased urinary zinc excretion were balanced by lower (p<0.05) endogenous excretion of zinc as evidenced by reduced excretion of Zn65 in feces during the second 4-day period. The mean biological half-life of Zn65 tended to be lower in patients with pancreatic insufficiency as compared to alcoholic control subjects, however the difference did not reach statistical significance. CONCLUSION: These observations indicate marked alterations in zinc metabolism in patients with advanced chronic pancreatic disease and provide greater insight into development of zinc deficiency in this group of patients. PMID- 9853536 TI - Protein-energy undernutrition and the risk of mortality within six years of hospital discharge. AB - OBJECTIVE: The primary objective was to determine whether protein-energy undernutrition among elderly patients discharged from the hospital remains a significant risk factor for mortality beyond 1 year. DESIGN: Prospective Survey (cohort study). SETTING: Outpatient follow-up of patients discharged from a Geriatric Rehabilitation Unit (GRU) of a Veterans Administration hospital. PARTICIPANTS: Of 350 randomly selected admissions to the GRU, 322 were discharged alive from the hospital. These 322 patients represented the study population of whom 99% were male, and 75% were white. The average age of the study patients was 76 (range 58 to 102) years. MEASUREMENTS: At admission and again at discharge, each patient completed a comprehensive medical, functional, neuro-psychological, socioeconomic, and nutritional assessment. Subsequent to discharge, each subject was tracked for an average of 6 years. In addition to including serum albumin and other putative nutrition indicators in the data set, a "nutrition-risk" indicator variable was created. Subjects were stratified into the nutrition "high-risk" group if their albumin was less than 30 g/L or BMI was less than 19; and, "low risk" group if albumin was equal to or greater than 35 g/L and BMI equal to or greater than 22. All others represented the "moderate-risk" group. RESULTS: Within the 6-year post-hospital-discharge follow-up period, 237 study subjects (74%) died. Based on the Cox proportional hazards survival model, the variable most strongly associated with mortality was discharge "nutrition-risk" followed by the Katz Index of ADL Score, diagnosis of congestive heart failure, discharge location (home vs. institution), age, and marital status. Within the first 4.5 years of follow-up, the relationship between "nutrition-risk" and mortality remained constant. After 4.5 years, the strength of the correlation began to diminish. CONCLUSIONS: Among the elderly, protein-energy undernutrition present at hospital discharge appears to be a strong independent risk factor for mortality during the subsequent 4.5 years or longer. PMID- 9853535 TI - Current zinc intake and risk of diabetes and coronary artery disease and factors associated with insulin resistance in rural and urban populations of North India. AB - OBJECTIVE: To determine the association between current zinc intake and prevalence of coronary artery disease (CAD) and diabetes as well as factors associated with insulin resistance. DESIGN, SUBJECTS AND METHODS: In this cross sectional survey, 3575 subjects, aged 25 to 64 years, including 1769 rural (894 men. 875 women) and 1806 urban (904 men, 902 women) subjects were studied. The survey methods included questionnaires for 7-day food intake record, physical examination, and electrocardiography using World Health Organization criteria. RESULTS: The prevalence of CAD, diabetes and glucose intolerance was significantly higher among subjects consuming lower intakes of dietary zinc. There was a higher prevalence of hypertension, hypertriglyceridemia and low high density lipoprotein cholesterol levels which showed significant upward trend with lower zinc intakes. Serum lipoprotein (a) and 2-hour plasma insulin levels also were associated with low zinc intake. Multivariate logistic regression analysis after adjustment for age showed that zinc intake and CAD were inversely associated. Serum zinc (odds ratio:men 0.77, women 0.57), serum triglycerides (men 0.86, women 0.81), blood pressure (0.83 men, women 0.76), diabetes mellitus (men 0.90, women 0.85), central obesity (men 0.88, women 0.87), glucose intolerance (men 0.66, women 0.57) and low high-density lipoprotein cholesterol (men 0.72, women 0.70) were significant risk factors for CAD (explained by tertiles of zinc status) in urban subjects. These associations were not observed in rural subjects. CONCLUSION: Lower consumption of dietary zinc and low serum zinc levels were associated with an increased prevalence of CAD and diabetes and several of their associated risk factors including hypertension, hypertriglyceridemia and other factors suggestive of mild insulin resistance in urban subjects. PMID- 9853537 TI - Nutrient intake and food group consumption of 10-year-olds by sugar intake level: the Bogalusa Heart Study. AB - OBJECTIVE: The effect of total sugar intake on nutrient intake and food group consumption was examined in children. METHODS: Twenty-four hour dietary recalls were collected on a sample of 568 ten-year-olds from two cross-sectional surveys. The population was stratified according to total sugar intake quartiles: < or = 25th (46 g/1000 kcal), 25-50th (67 g/1000 kcal), 50-75th (81 g/1000 kcal) and > or = 75th (106 g/1000 kcal). RESULTS: No significant difference was shown in energy intakes across the quartiles and no significant race or gender differences were observed in mean total sugar intakes. However, with increasing total sugar intake, there was a significant linear decrease in mean intakes of protein, fat, saturated fat, starch, cholesterol, sodium, vitamins B6 and E, thiamin, niacin, iron, and zinc; and a significant linear increase in mean intakes of carbohydrate, fructose, lactose, sucrose, vitamin D, and calcium. Eating patterns reflected the differing nutrient intakes, with high sugar consumers having significantly higher intakes of total g of candy, beverages and milk and lower intakes of total g of meats, and cheese than lower sugar consumers. The nutritional quality of children's diets high in total sugar appear to be adequate regarding vitamin and mineral intakes and are closer to meeting current dietary fat recommendations. PMID- 9853538 TI - Effects of dietary n-6 and n-3 lipids on antioxidant defense system in livers of exercised rats. AB - OBJECTIVE: The present study was designed to investigate the effects of dietary n 6 and n-3 lipids and exercise on the activities of hepatic antioxidant enzymes and microsomal lipid composition and peroxidation in Fischer-344 male rats. METHODS: Weanling male Fischer-344 rats were fed ad libitum semipurified diets containing 10% corn oil (CO) or 10% fish oil (FO), with equal levels of antioxidants. After 2 months on the diets, weight-matched animals in each diet group were divided into sedentary (S) and exercised (Ex) groups, and the diets were continued. The animals in the exercise group were run on a treadmill 30 to 40 minutes to exhaustion 6 days/week for 2 months. At the end of 2 months, the rats were sacrificed and livers were collected; antioxidant enzymes were determined in the cytosol, fatty acid composition was analyzed in the microsomes, and vitamin E levels were analyzed in the sera. RESULTS: The rats in the FO-S group exhibited significantly higher liver cytosolic catalase activity, while their superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were significantly lower compared to the CO-S group. The GSH-Px activity was significantly higher in the FO-Ex group compared to FO-S group. The source of dietary lipids significantly influenced the fatty acid composition of the total lipids in the microsomes. Feeding the FO-based diet significantly increased 18:0 and n-3 fatty acids incorporation into the microsomes (18:3, 20:5, 22:5, and 22:6), whereas ingestion of CO resulted in a significant increase in 14:0, 14:1, 18:1, and n-6 fatty acids (18:2 and 20:4). The serum vitamin E levels were significantly higher in the CO groups, and exercise had no effect on vitamin E levels. Exercise significantly decreased the generation of thiobarbituric acid reactive substances (TBARS) by liver microsomes. Consumption of FO, which is highly susceptible to oxidation, did not show any significant changes in membrane lipid peroxidation. CONCLUSIONS: The present study suggests that feeding FO increases the activity of liver cytosolic catalase in FO-S rats and GSH-Px in FO Ex rats. In addition, exercise significantly decreased the generation of TBARS by the liver microsomal lipids. Serum vitamin E levels were higher in the CO group and exercise did not alter vitamin E levels. This suggests that the amount of vitamin E included in the diets was possibly adequate to cope with the oxidative stress induced during exercise. PMID- 9853539 TI - Utility of a short-term 25% carbohydrate diet on improving glycemic control in type 2 diabetes mellitus. AB - OBJECTIVE: To determine if introduction of a low carbohydrate diet might be a useful option for type 2 diabetic patients who do not achieve glucose target levels despite conventional treatment. METHODS: Subjects with type 2 diabetes, either treated with diet alone (n=9) or second generation sulfonylurea agents (n= 19), which were discontinued, were placed on a diet based on ideal body weight and comprised of 25% carbohydrate. After a mean of 8 weeks, they were then switched to a caloricly equivalent diet, but composed of 55% carbohydrate. RESULTS: Compared to baseline diet, after 8 weeks of a 25% diet, subjects showed significantly improved glycemia as evidenced by fasting blood glucose values (p<0.005) and hemoglobin A1c levels (p<0.05). Those previously treated with oral hypoglycemic agents showed, in addition, a significant decrease in weight and diastolic blood pressure despite the discontinuation of the oral agent. When then placed on a 55% carbohydrate diet, the hemoglobin A1c rose significantly over the ensuing next 12 weeks (p<0.05). CONCLUSION: A low carbohydrate, caloricly restricted diet has beneficial short-term effects in subjects with type 2 who have failed either diet or sulfonylurea therapy and may obviate the necessity for insulin. Our study also affirms the need for reassessing the role of diet whenever type 2 diabetic patients manifests hyperglycemia, despite conventional oral treatment or diet management. PMID- 9853540 TI - Cookies enriched with psyllium or oat bran lower plasma LDL cholesterol in normal and hypercholesterolemic men from Northern Mexico. AB - BACKGROUND: Psyllium and oat bran have been shown to lower plasma LDL cholesterol levels in different populations. Hypercholesterolemia is prevalent in the Northern part of Mexico and might be associated to dietary habits and sedentary lifestyle. METHODS: Sedentary normal (cholesterol<200 mg/dL) (n=36) and hypercholesterolemic (cholesterol>220 mg/dL) (n=30) men from the Northern part of Mexico aged 20 to 45 years of age participated in an 8-week study to determine the effects of dietary soluble fiber, either psyllium or oat bran, in lowering plasma LDL cholesterol in this population. Fiber was administered by feeding the subjects an amount of cookies (100 g) equivalent to 1.3 or 2.6 g/day of soluble fiber from psyllium or oat bran, respectively. Subjects were randomly allocated to three groups: a control group consuming cookies with wheat bran, a known source of fiber with no cholesterol lowering effects, psyllium, or oat bran. RESULTS: Food frequency questionnaires indicated that subjects from the three groups had similar intakes of foods classified as hypercholesterolemic (p>0.05). Plasma LDL cholesterol concentrations were reduced by an average of 22.6 and 26% in the psyllium and oat bran groups (p<0.001) while a non-significant reduction of 8.4% was observed in the hypercholesterolemic individuals from the control group. No effects on plasma HDL or triglycerides levels were observed among the three dietary treatments except for hypercholesterolemic individuals supplemented with oat bran where a 28% reduction in plasma triglycerides was observed after 8 weeks (p<0.01). CONCLUSION: These results indicate that psyllium and oat bran are efficacious in lowering plasma LDL cholesterol in both normal and hypercholesterolemic individuals from this population. PMID- 9853541 TI - Physiological effects of resistant starches on fecal bulk, short chain fatty acids, blood lipids and glycemic index. AB - OBJECTIVE: To assess the effects on fecal bulking, fecal short chain fatty acid (SCFA) production, blood lipids and glycemic indices of two different forms of resistant starch (RS2 and RS3) from a high-amylose cornstarch. METHODS: Twenty four healthy subjects (12 men; 12 women) consumed four supplements taken for 2 weeks in random order separated by 2-week washout periods. The supplements were a low-fiber (control) and supplements providing an additional 30 g dietary fiber as wheat bran (high-fiber control) or the equivalent amount of resistant starch analyzed gravimetrically as dietary fiber from RS2 or RS3. Four-day fecal collections and 12-hour breath gas collections were obtained at the end of each period. Fasting blood was taken at the beginning and end of each period. Glycemic indices of supplements were also assessed. RESULTS: The wheat bran supplement increased fecal bulk 96+/-14 g/day compared with the low-fiber control (p<0.001) with the mean for both resistant starches also being greater (22+/-8 g/day) than the low-fiber control (p=0.013). On the resistant starch phases, the mean fecal butyrate:SCFA ratio, which has been suggested to have positive implications for colonic health, was significantly above the low-fiber control by 31+/-14% (p=0.035). Resistant starches did not alter serum lipids, urea or breath H2 or CH4. No significant differences in glycemic index were seen between the RS and control supplements. CONCLUSION: The potential physiological benefits of the resistant starches studied appear to relate to colonic health in terms of effects on fecal bulk and SCFA metabolism. PMID- 9853542 TI - Effect of UV exposure and beta-carotene supplementation on delayed-type hypersensitivity response in healthy older men. AB - OBJECTIVES: The aims of this study were to determine if ultraviolet light (UV) is immunosuppressive in healthy older males, if beta-carotene (betaC) supplementation could prevent any observed UV-induced immunosuppression, and to compare these effects with those observed previously in younger men. METHODS: The study was a placebo-controlled, randomized trial that employed a 2 x 2 factorial design. Healthy older men (mean age 65.5 years) received 30 mg betaC or placebo daily throughout the 47-day trial, while on a low carotenoid diet. After 28 days, half of each group received 12 suberythemic exposures to UV over a 16-day period. Delayed-type hypersensitivity (DTH) tests and plasma carotenoid assays were performed at baseline, pre-UV and post-UV time points, with DTH testing performed on an area of skin protected from UV exposure. RESULTS: UV exposure resulted in significantly suppressed DTH response in the placebo group but not in the betaC UV group. While there was no significant interaction between betaC supplementation and UV on DTH response, there was a significant inverse relationship between final plasma betaC concentration and extent of UV-induced suppression of DTH response. A similar correlation existed among subjects not exposed to UV. CONCLUSIONS: Suberythemic UV exposure was immunosuppressive, as measured by DTH response, in healthy older men as in younger men. Higher plasma betaC was significantly associated with maintenance of DTH response, although the extent of protective effect of betaC appeared less than previously observed in younger subjects. The attenuated effect of betaC in the older UV-exposed subjects may have resulted in part from muted plasma betaC responses to betaC supplementation and/or higher plasma vitamin E levels than those of younger men. The finding that stronger DTH responses were associated with higher plasma betaC concentrations in both UV and non-UV subjects further supports a role for this nutrient in immunomodulation. PMID- 9853543 TI - Absorption of dietary and supplemental folate in women with prior pregnancies with neural tube defects and controls. AB - BACKGROUND: The Public Health Service of the United States recommends that all women capable of childbearing consume .4 mg (400 microg) folic acid per day to decrease the risk of having a pregnancy affected by a neural tube defect such as spina bifida or anencephaly. Three strategies are available to women to achieve this goal: use of dietary supplements; use of fortified foods; and/or increased intake of naturally occurring folate from foods. Identification of the most effective vehicle for delivery of folate to all women is critical in order to prevent these devastating congenital defects. OBJECTIVE: To investigate the difference in response to an oral load of folate both from naturally occurring food sources and synthetic supplements among women with prior pregnancies affected by neural tube defects and controls. METHODS: We compared the absorption of test doses of 400 microg pteroylglutamic acid (unconjugated or synthetic folic acid found in supplements) and 400 microg pteroylpolyglutamic acid (conjugated or food folate) in 10 women with a history of neural tube defect affected pregnancies and eight controls with normal birth outcomes. The folate test dose was given as either 32 fluid ounces of orange juice or a folic acid single supplement pill. All participants received each test dose at separate clinic visits. The response to each test dose was measured by constructing an area under the curve (AUC) from the serum folate levels at 1, 2 and 3 hours post dose and applying a t-test to compare within and between cases and controls. We also compared red cell folate, vitamin B12, zinc and homocysteine between cases and controls. RESULTS: Within group comparisons showed that the area under the curve was significantly greater for the pteroylglutamic acid dose compared to the pteroylpolyglutamic acid dose for both cases and controls (p=0.02 and p=0.03, respectively). In a between group comparison, control women had a greater serum folate response to both forms of the vitamin compared to the case women, but the difference reached statistical significance only for the pteroylglutamic acid dose (p=0.02). Other measured nutrients differed between cases and controls, but did not reach statistical significance. CONCLUSION: We conclude that for all women synthetic folic acid as supplements or fortified foods may be the best way to increase acute folate levels in the blood, and thus delivery to the developing embryo. Further, since case women had a diminished response to both forms of the vitamin, and some case women had almost no response, we speculate that women with prior affected pregnancies may need a larger dose of folate to elicit a plasma response equivalent to the general population. PMID- 9853544 TI - Lactose intolerance and self-reported milk intolerance: relationship with lactose maldigestion and nutrient intake. Lactase Deficiency Study Group. AB - BACKGROUND: The relationship between lactose-maldigestion, self-reported milk intolerance and gastrointestinal symptoms has not been clearly defined. OBJECTIVES: To evaluate: a) the prevalence of lactose maldigestion and lactose intolerance in a sample of the general population taken from a rural center; b) the frequency of self-reported milk-intolerance and its correlation with lactose maldigestion; c) the influence of lactose maldigestion, lactose intolerance and self-reported milk intolerance on dietary habits and consumption of total calories, protein, and calcium. SUBJECTS: We studied a randomized sample of the general population in a small center in Sicily. 323 subjects (150 males, 173 females), age range 5 to 85 years (median 44) were included and underwent H2 breath test after 25 g lactose load. The preliminary dietary investigation spanned 7 consecutive days using a printed dietary form and was under the daily control of a team of dietitians. METHODS: The dietary investigation was completed in the first part of the study and the results were analyzed for nutrient composition by a computerized database. The subjects were then divided into self reported milk-intolerants and self-reported milk-tolerants and they underwent H2 breath testing; subjects with H2 concentration >20 ppm over the baseline concentration were considered maldigesters and those with one or more symptoms were classified as intolerants. RESULTS: 104/323 subjects (32.2%) were lactose maldigesters but tolerants, while 13/323 (4%) were lactose maldigesters and intolerants. In each age-class group (pediatric, adult, and elderly subjects) only the lactose maldigester and intolerant subjects showed differences in nutrient intake with a significantly lower daily consumption of milk and a lower calcium intake. 49/323 subjects were self-reported milk-intolerants; of these, 26 (53%) were lactose maldigesters but tolerants, 18 (37%) were lactose digesters and tolerants and only 5 (10%) were lactose maldigesters and intolerants. In the whole group of self-reported milk-intolerants, dietary milk consumption was significantly reduced and calcium intake was lower than in all the other subjects studied (320 mg/day vs. 585 mg/day, p<0.05). CONCLUSIONS: In studies of the general population, the frequency of lactose intolerance is much lower than that of lactose maldigestion. Gastrointestinal symptoms after lactose load in self reported milk-intolerants are found in only a very low number of these subjects. Furthermore, in these subjects we observed an unnecessary reduction in milk consumption and an insufficient dietary calcium intake. PMID- 9853545 TI - Nutritional rickets in suburbia. AB - OBJECTIVE: Vitamin D deficiency continues to be a problem in pediatrics. This report presents four children, one Caucasian male and three African-American females aged 4 to 24 months who were treated for vitamin D deficiency rickets. METHODS: One female was diagnosed in the Emergency Department during evaluation of a viral syndrome, another presented with hypocalcemic seizures and the third was a self-referral for evaluation of widened wrists. The male had biochemical rickets discovered incidentally during a hospitalization for pneumonia. All were breastfed without formula supplements. The 24-month female had severe cow and soy protein allergies and received multivitamin supplements intermittently. Birth order was from third to sixth child. Two families practiced Islam and the mothers wore veils. The females had a weight deficit for height. The females demonstrated a rachitic rosary, widening of the wrists and leg bowing. At diagnosis the serum calcium was 5.0-8.6 mg/dl, the inorganic phosphorus was 1.5-3.9 mg/dl and the alkaline phosphatase was 408-3324 U/L. The serum intact parathormone levels and the vitamin D levels were measured at Nichols Laboratories. The 25-OH vitamin D levels were 2-22 ng/ml and the 1,25(OH)2 vitamin D levels were 14-122 pg/ml. All had elevated parathormone levels. The three females had roentgenographic evidence of rickets. Two of the children also demonstrated iron deficiency. RESULTS: All patients responded to Vitamin D supplements, beginning at 2000 IU for the male and 8,000-10,000 IU daily for the females. Two children were also given calcium supplements. The three females all showed complete healing of the rickets radiologically within six months. The serum intact parathormone demonstrated an inverse correlation with the serum calcium during recovery (r=-0.669; p<0.05). CONCLUSION: Vitamin D deficiency does still occur. Breastfed children of multiparous mothers, with increased skin pigmentation, living in the higher latitudes are at increased risk and would benefit from vitamin D supplementation while breastfeeding. PMID- 9853547 TI - Carnitine may benefit athletes. PMID- 9853546 TI - Calcium and phosphorus in milk of Brazilian mothers using oral contraceptives. AB - OBJECTIVE: Oral contraceptives (OC) are the most efficient method of contraception and it is the most prescribed by doctors in developing countries. Therefore we studied the effects of combination pill and mini-pill on calcium and phosphorus in milk of breast-feeding mothers at different stages of lactation. METHODS: Fifty-four breast-feeding mothers made up three study groups: 33 mothers who had been advised by their doctors to use either combination pill (12), or mini-pill (21), as well as a control group of 21 mothers that used no hormonal contraceptives. All mothers completed a questionnaire and provided samples of milk before and after a measured period of observation. Mean duration of study was 76, 120, and 101 days, respectively for users of mini-pill, combination pill, and controls. Determination of calcium and phosphorus was done by inductively coupled plasma-atomic absorption spectrometry. RESULTS: Overall the decrease in milk concentrations of phosphorus (6%) and calcium (26.3%) during the study period was not influenced by OC treatment. Regression analyses which took into consideration length of treatment, socioeconomic status, number of children, duration of previous lactation, type of contraceptive, and age of mothers and repeated measurements (before and after OC) showed that milk calcium was significantly affected by stage of lactation (p=0.0013). CONCLUSION: The use of hormonal contraceptive such as the combination pill (levonorgestrel 0.15 mg+ethynilestradiol 0.03 mg) and mini-pill (norethindone 0.35 mg) does not seem to affect the secretion of calcium and phosphorus in milk of mothers. PMID- 9853548 TI - Predicting basal metabolic rate in obese children. PMID- 9853549 TI - Guide for judicious use of the paracetamol absorption technique in a study of gastric emptying rate of liquids. AB - The paracetamol absorption technique, a widely used method for evaluating the gastric emptying rate of liquids, appears to be performed inappropriately, resulting from a lack of consideration of pharmacokinetics in paracetamol absorption. This review suggests that appropriate study designs and logical choice of the parameters for the rate of paracetamol absorption are the cornerstone of reliable investigation of gastric emptying using the paracetamol method. PMID- 9853550 TI - Nitric oxide: a regulator of mucosal defense and injury. AB - There is an abundance of evidence that nitric oxide plays a critical role in regulating several components of gastrointestinal mucosal defense. Suppression of nitric oxide synthesis increases susceptibility to injury, while administration of nitric oxide donors increases resistance to injury. On the other hand, nitric oxide has been implicated as a mediator of tissue injury in the gastrointestinal tract during inflammatory reactions. In these cases, the nitric oxide is generally believed to be derived from an inducible isoform of nitric oxide synthase. In this review, we provide an overview of the evidence for and against these dual roles of nitric oxide in modulating gastrointestinal mucosal defense and injury. We also highlight the potential therapeutic benefits that may be realized through modulation of tissue nitric oxide levels. PMID- 9853551 TI - Genetic changes in colorectal adenoma and cancer in relation to various morphologic aspects. PMID- 9853552 TI - Expression of sulfated carbohydrate chain and core peptides of mucin detected by monoclonal antibodies in Barrett's esophagus and esophageal adenocarcinoma. AB - Columnar epithelium-lined esophagus (Barrett's esophagus) is an acquired disorder associated with a high incidence of adenocarcinoma of the lower esophagus. Columnar epithelium resembling intestinal metaplasia (IM) is especially important, since it is considered to be a premalignant condition. The aim of this study was to define the sulfated carbohydrate chain of mucin and its core peptide profile in Barrett's esophagus (BE) and to compare it with the profile in Barrett's adenocarcinoma and lower esophageal adenocarcinoma. The sulfated carbohydrate chain was not expressed in 16 specimens of normal esophageal epithelium, but in BE, it was expressed in 50% (8/16) of the specimens. This chain was detected in 100% (7/7) of esophageal adenocarcinoma specimens, including four cases of Barrett's adenocarcinoma. These data suggest that the sulfated carbohydrate chain may be associated with malignant phenotype of the esophagus. MUC1 core peptide was positive in 87% (13/15) of BE specimens and in 29% (2/7) of the esophageal adenocarcinoma specimens. MUC2 core peptide was present in 57% (8/14) and 43% (3/7) of these specimens, respectively. These data suggest that Barrett's epithelium, which is similar to IM, but not normal esophageal epithelium, expresses the sulfated sugar chain which is known to be present in gastric IM and colonic mucosa. However, there was no significant correlation between the expression of the sulfated sugar chain and the expression of either mucin core peptide MUC1 or MUC2. Thus, this carbohydrate chain may be expressed on as yet unidentified core proteins, other than MUC1 or MUC2 core peptide, in BE and esophageal adenocarcinoma. Identification of these proteins may be very important in helping to detect premalignant status in BE. PMID- 9853553 TI - Expression of manganese superoxide dismutase in esophageal and gastric cancers. AB - The tumor-killing activity of radiotherapy and chemotherapy for cancer is closely associated with the production of active oxygen, and the relation between therapeutic resistance and active oxygen scavengers produced by the tumor itself is gaining more attention. It is considered that manganese superoxide dismutase (MnSOD) protects host cells from oxidative stress, in synergy with other antioxidant enzymes. In this study, we used a quantitative polymerase chain reaction assay to measure MnSOD mRNA in resected specimens from patients with esophageal and gastric cancers. In both esophageal and gastric cancers, the level of MnSOD mRNA was significantly elevated in cancer tissue compared to non-cancer tissue (P < 0.01). In gastric cancer tissue, the MnSOD mRNA level was significantly higher than in esophageal cancer tissue (P < 0.01). The significance of MnSOD in cancer tissue was investigated further by measuring MnSOD content in resected specimens using an enzyme-linked immunosorbent assay, and by examining its location by an immunohistochemical method. Upregulation of MnSOD in cancer tissue most likely serves as a protective mechanism against anti cancer therapies known to produce superoxide radicals as a key component of their tumor-killing activity. PMID- 9853554 TI - Effects of trimebutine maleate on gastric motility in patients with gastric ulcer. AB - The effects of trimebutine maleate (TM), a prokinetic drug, on gastrointestinal motility in patients with gastric ulcer were investigated. Twenty patients with active gastric ulcers were allocated to two groups; 10 patients received a proton pump inhibitor alone (PPI group), given orally, and 10 patients received oral TM in combination with a PPI (PPI + TM group), each for a period of 8 weeks. Electrogastrography (EGG) and gastric emptying were measured before and after the treatment period. During the active ulcer stage, tachygastria (more than 0.06 Hz) or bradygastria (less than 0.04 Hz) in the EGG frequency were observed in 9 patients either before or after meals. During the healed ulcer stage, tachygastria or bradygastria was observed in 4 of 10 patients in the PPI group, while in the PPI + TM group, 1 patient had tachygastria and none had bradygastria. Postprandial dip (PD) was observed in 3 of the 20 patients during the active stage, while after treatment, PD was observed in 3 patients in the PPI group and in 6 patients in the PPI + TM group, respectively. Gastric emptying in the PPI group did not show any change between before and after treatment, while that in the PPI + TM group improved significantly after treatment. These results suggest that TM may have an ameliorative effect on abnormal gastric motility in patients with gastric ulcer. PMID- 9853555 TI - Adrenomedullin in the gastrointestinal tract. Distribution and gene expression in rat and augmented gastric adrenomedullin after fasting. AB - The aim of this study was to investigate the regional distribution, molecular forms, and gene expression of adrenomedullin in the rat gastrointestinal tract and to examine physiological changes in gastric adrenomedullin after 24-h fasting. The tissue concentration was measured by radioimmunoassay. The molecular forms were analyzed by high performance liquid chromatography. mRNA levels were quantified by Northern blotting and cells positive for adrenomedullin immunoreactivity were localized by immunohistochemistry. A high concentration of adrenomedullin was found in stomach, cecum, and colon (450-520 fmol/g wet tissue). Adrenomedullin immunoreactivity was also detected in duodenum, jejunum, and ileum (200-250 fmol/g wet tissue). Transcripts of the adrenomedullin gene were widely expressed throughout the gastrointestinal tract. The major form of adrenomedullin immunoreactivity in stomach and colon corresponded precisely with authentic adrenomedullin peptide. Adrenomedullin immunoreactive cells were present in the gastrointestinal endocrine system. The concentration and mRNA level of gastric adrenomedullin after fasting were significantly increased compared with findings in controls. Adrenomedullin is ubiquitous in the gastrointestinal tract, and may be produced by endocrine cells. The results suggest that adrenomedullin, through its potent vasodilating activity, may play some role, in the stomach including the regulation of the mucosal blood flow. PMID- 9853556 TI - Mechanism of inhibitory effect of glucagon on gastrointestinal motility and cause of side effects of glucagon. AB - Glucagon is commonly used during gastrointestinal examinations for the temporary inhibition of gastroduodenal movements. Three preparations of glucagon are now clinically available: those prepared by extraction from the pancreas (GL-P), by chemical synthesis (GL-S), and by genetic recombination (GL-G). The aim of this study was examine the mechanism of the inhibitory effect of glucagon on gastrointestinal motility and the cause of its side effects by comparing three glucagon preparations. In four conscious dogs, gastrointestinal contractions were monitored by means of chronically implanted force transducers. Each glucagon preparation (GL-P [15 microg/kg], GL-S [5, 15, 45 microg/kg], GL-G [15 microg/kg]), scopolamine butylbromide (0.4 mg/ kg), or saline was administered intravenously 20 min after the termination of spontaneous phase III contractions, and blood samples were taken at 5- to 10-min intervals. Barium was administered into the stomach 10 min after the infusion of each drug. The arrival of a barium meal in the stomach immediately stimulated gastrointestinal contractions, and the barium meal was expelled into the duodenum and jejunum from the stomach. Intravenous injection of 15 microg GL-S first stimulated duodenal contractions that propagated to the jejunum, followed by strong inhibition of the barium induced gastrointestinal contractions. This inhibitory effect of glucagon and the activity of the glucagon-induced duodenal contractions were dose-related. The inhibitory effects of GL-G and GL-S were stronger than that of GL-P. Blood glucose and plasma insulin concentrations were raised after intravenous injection of each glucagon preparation, but there was no difference among the three preparations and no dose relationship. The inhibitory effects of glucagon depend on the material purity and dose, and the inhibitory mechanism was independent of any effect on carbohydrate metabolism. Glucagon administration caused phase III like contractions in the duodenum and jejunum, which may be responsible for the side effects of glucagon. PMID- 9853557 TI - Characteristics of two cancer cell lines derived from metastatic foci in liver and peritoneum of a patient with colon cancer. AB - Two cancer cell lines were established in vitro from a single patient with colon cancer; AKT-CC-K-LM cells from liver metastatic nodules and AKT-CC-K-PC cells from peritoneal dissemination nodules. The two cell lines were similar in doubling time, number of chromosomes, and chromosomal abnormalities. However, they differed in morphology in vitro, in the expression level of cell surface adhesion molecules (carcinoembryonic antigen; CEA, E-cadherin, sialyl Le(a), sialyl Le(x), and CD44v6), and in their metastatic properties. AKT-CC-K-LM cells grew in vitro as adherent clusters and AKT-CC-K-PC cells as adherent single cells. The expression levels of CEA, E-cadherin, sialyl Le(a), and sialyl Le(x) was significantly higher in AKT-CC-K-LM cells. The expression of CD44v6 was significantly higher in AKT-CC-K-PC cells. After the injection of AKT-CC-K-LM cells to the spleen or peritoneal cavity of severe combined immune deficiency mice, metastatic nodules were observed only in the liver. In contrast, the injection of AKT-CC-K-PC cells to the spleen or peritoneal cavity yielded metastatic nodules only in the peritoneal cavity. These cell lines may contribute to elucidating the relationship between cell surface adhesion molecules and the metastatic properties of cancer cells. PMID- 9853558 TI - Prevalence and clinical features of hepatitis delta virus infection in the Miyako Islands, Okinawa, Japan. AB - The aims of this study were twofold: (1) to determine the prevalence and clinical features of hepatitis delta virus (HDV) infection among subjects positive for hepatitis B surface antigen (HBsAg) living in the Miyako Islands, Okinawa Prefecture, Japan, and (2) to clarify the relationship between HDV-RNA level and severity of HDV-related liver disease. One hundred and ninety-nine HBsAg-positive subjects (123 asymptomatic carriers [ASCs], 3 patients with acute hepatitis [AH], 50 patients with chronic hepatitis [CH], 15 patients with liver cirrhosis [LC], and 8 patients with hepatocellular carcinoma [HCC], were tested for antibody to HDV (anti-HDV) by radioimmunoassay. Anti-HDV-positive individuals were examined to determine semi-quantified HDV-RNA level by polymerase chain reaction (PCR). The overall prevalence of anti-HDV among the 199 subjects was 21.1%. The positivity rate tended to increase with age or the severity of the underlying liver disease: anti-HDV-positive rates were 10.6% (13/123) in ASCs, 32.0% (16/50) in patients with CH, 40.0% (6/15) in patients with LC, and 87.5% (7/8) in patients with HCC. None of the patients with AH were positive for anti-HDV. There was no correlation between semi-quantified serum HDV-RNA levels and the severity of chronic liver disease in patients positive for anti-HDV. The present study showed the local spread of HDV infection in the Miyako Islands, Okinawa, Japan. Although the anti-HDV positivity rate tended to increase with the severity of the underlying liver disease, the severity of HDV-related liver disease did not correlate with the semi-quantified serum HDV-RNA level. PMID- 9853559 TI - Suppressive effect of ethanol on the expression of hepatic asialoglycoprotein receptors augmented by interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha. AB - Blood levels of inflammatory-related cytokines, including interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)-alpha, are elevated in patients with alcoholic liver diseases. We investigated the effects of these cytokines and ethanol on the expression of hepatic asialoglycoprotein receptors (AGPRs) in a human hepatoblastoma cell line, HepG2. An [125I]-asialo-orosomucoid binding assay showed significant increases in surface AGPR numbers in HepG2 cells by treatment with IL-1beta, IL-6, and TNF-alpha, to levels which were approximately 130% of the values in untreated control cells. However, the enhanced AGPR numbers induced by treatment with these cytokines were markedly suppressed, to 70%-80% of the number in the untreated cells, by treatment with ethanol. Immunological detection of AGPR with a specific antibody demonstrated that the modulation of surface AGPR numbers was correlated with the cellular expression levels of AGPR. These results suggest that, although IL-1beta, IL-6, and TNF-alpha stimulate the synthesis of hepatic AGPR, ethanol suppresses the expression of AGPR augmented by these cytokines. This leads to an increase in serum asialo-orosomucoid levels caused by the disordered catabolism mediated by AGPR in patients with alcoholic liver disease. PMID- 9853560 TI - Vector manometric study of the sphincter of Oddi in the dog: functional and morphological correlation. AB - The relationship between sphincter of Oddi pressure and the morphological structure of the sphincter was studied in eight dogs prepared with a duodenal cannula. Sphincter of Oddi manometry was performed in awake animals in three directions, ventral, left dorsal, and right dorsal, using a catheter with three radial side holes for recording at one level. The pressure in the ventral direction (26.6+/-1.06 mmHg) (mean+/-SEM) was significantly lower than that in the left and right dorsal directions (30.6+/-1.42 and 31.2+/-1.23 mmHg, respectively). This functional manometric difference in the three directions correlated closely with the morphological structure of the sphincter of Oddi; the sum of the thickness of the sphincter of Oddi muscle and duodenal proper muscle was greater on the dorsal than on the ventral side. To our knowledge, this is the first report of axial asymmetry in sphincter of Oddi pressure. PMID- 9853561 TI - Complete response of early gastric cancer to uracil and tegafur. AB - A 74-year-old Japanese woman with early gastric cancer was successfully treated with uracil and tegafur (UFT). She was diagnosed by endoscopy (including endoscopic biopsy and endosonography) with an early gastric cancer, type IIa + IIc, on the greater curvature of the angulus. Surgical procedures or endoscopic therapy could not be performed because the patient had severe ischemic heart disease. Therefore, chemotherapy with UFT was administered at 300 mg/day for 15 months. Follow-up endoscopy, endosonography, and biopsy showed disappearance of the gastric cancer. To our knowledge, this is the first case report of the complete response of an early gastric cancer to UFT in the English-language literature. PMID- 9853562 TI - Appendico-ileo-vesical fistula. AB - Appendico-vesical fistula is a rare condition. In total, 109 cases, most secondary to appendicitis, have been reported in the English-language literature. We report the first case, to our knowledge, of appendico-ileo-vesical fistula secondary to appendiceal diverticulitis. An enterovesical fistula was diagnosed by urine culture, cystoscopy, and computed tomography. The locations of enteric opening sites were demonstrated by barium enema and colonoscopy. Ileocecal resection and fistulectomy with primary reconstruction were performed. We believe that accurate pre- and intra-operative diagnosis is essential for cure. This case demonstrates the importance of barium enema and colonoscopic examinations in the diagnosis and treatment of complicated enterovesical fistula. PMID- 9853563 TI - A schwannoma of the cecum: case report and review of Japanese schwannomas in the large intestine. AB - A 66-year-old Japanese man had a positive fecal occult blood test at a regular check-up, and a large polypoid mass was detected in the cecum by barium enema study. Colonoscopy showed a submucosal tumor with ulcer protruding into the cecal lumen. A large-forceps biopsy specimen was taken from the bottom of the ulcer. With the tentative diagnosis of neurogenic tumor, ileocecal resection was performed. The tumor showed spindle-cell proliferation in a concentric or fascicular pattern. Immunohistochemically, the tumor cells were diffusely positive for S-100 protein, and they had intracytoplasmic periodic acid Schiff (PAS)-positive crystalloids. The mitosis count was low (about 1 per 20 high-power fields). The pathological diagnosis of this tumor was benign gastrointestinal schwannoma. A large number of schwannoma cases have been reported since 1910 when Verocay reported it as a true tumor that stemmed from Schwann cells and did not contain neuroganglion cells. However, gastrointestinal schwannomas are rare, and schwannomas of the large intestine are extremely rare. We reviewed 40 cases already reported in Japan and this present case in order to clarify the clinicopathological features of this tumor. PMID- 9853564 TI - Zinc deficiency manifested by dermatitis and visual dysfunction in a patient with Crohn's disease. AB - We report a case of Crohn's disease with low serum zinc concentration in a 26 year-old woman. She demonstrated acrodermatitis enteropathica and decreased visual acuity during total parenteral nutrition. Subsequent intravenous zinc supplementation resulted in alleviation of the skin lesions and improvement of visual acuity. This case supports the notion that depressed serum zinc in Crohn's disease may cause clinical manifestations, such as acrodermatitis enteropathica and retinal dysfunction, which may be correctable with zinc supplementation. PMID- 9853565 TI - Acute hepatic failure due to hepatosplenic B-cell non-Hodgkin's lymphoma in a patient infected with hepatitis C virus. AB - We report a 75-year-old Japanese man infected with hepatitis C virus (HCV) who died of acute hepatic failure due to the hepatic infiltration of B-cell non Hodgkin's lymphoma (NHL) cells. He suddenly developed jaundice, fatigue, fever, and hepatosplenomegaly during the course of chronic infection with HCV. Postmortem liver necropsy revealed extensive infiltration of lymphoma cells into the liver. Although the association between HCV infection and NHL has recently become a matter of concern, we believe this to be the first reported case of acute hepatic failure caused by hepatic involvement of non-Hodgkin's lymphoma in an HCV-infected patient. PMID- 9853566 TI - Hepatocellular carcinoma with obstructive jaundice successfully treated with a self-expandable metallic stent. AB - We report on a 71-year old man with hepatocellular carcinoma (HCC) whose obstructive jaundice was successfully treated with external irradiation and a self-expandable metallic stent (EMS); Wallstent; Schneider (Europe) AG, Bulach, Switzerland. He was admitted to our hospital because of jaundice. HCC was found in S8; the tumor had invaded the bile duct with growth in the common hepatic duct. Endoscopic nasobiliary drainage was performed with difficulty. Radiation therapy to the stenosis enabled us to place a Wallstent endoscopically. He survived without icterus for 1 year. PMID- 9853567 TI - Malignant insulinoma presenting a non-functioning metastatic liver tumor 14 years after resection of the primary tumor. AB - A 58-year-old woman who had undergone resection of insulinoma 14 year earlier visited our clinic complaining of abdominal discomfort. Computerized tomographic scan showed multiple liver tumors, and a diagnosis of metastatic tumor of malignant islet cell tumor was confirmed histologically. No oversecretion of hormones or hypoglycemic episode was observed on readmission. Thus, the insulinoma seemed to have transformed to a clinically non-functioning tumor. The patient was treated with transcatheter arterial embolization, resulting in clinical improvement with marked reduction in tumor size. Features of interest in this case included; (1) transformation to non-functioning metastatic liver tumor 14 years after resection of insulinoma, (2) the usefulness of transcatheter arterial embolization for multiple metastatic tumor of malignant islet cell tumor. PMID- 9853568 TI - Extrahepatic biliary cystadenocarcinoma arising from the left hepatic duct. AB - A 54-year-old man, who had no clinical symptoms, underwent a routine health checkup at our hospital. Abdominal ultrasonography disclosed a well demarcated tumor containing a solid portion occupying the dilated left hepatic duct and a cystic portion expanding into the parenchyma of the left hepatic lobe, with mild dilatation of the intrahepatic bile ducts. These findings were later confirmed by computed tomography (CT) and magnetic resonance imaging. Endoscopic retrograde cholangiography revealed a complete defect at the level of the left hepatic duct, while drip infusion cholangiographic-CT (DIC-CT) disclosed a defect of the left hepatic duct only, with the distal portions of the left intrahepatic ducts being visualized on the image. Hepatic angiography revealed light stains in the solid portion in the parenchymal phase. At left lobectomy, a multiloculated polyp-like tumor was found arising from the left hepatic duct and expanding into the parenchyma of the left hepatic lobe. Microscopically, all the lining cells in the cysts and the tumor cells in the solid portion showed the features of papillary adenocarcinoma. In this patient with extrahepatic biliary cystadenocarcinoma, DIC CT was useful in identifying the site of origin of the tumor, and hepatic angiography was also useful in differentiating this rare malignant tumor from benign cystadenoma. PMID- 9853569 TI - Survival of patient with late onset hepatic failure by living-related liver transplantation from maternal donor with incompatible blood type. AB - A 14-year-old girl with blood type B with late onset hepatic failure (LOHF) of unknown cause has survived through living-related liver transplantation (LRLT). No hepatitis virus, including HAV, HBV, HCV, and HGV, was positive at the onset of LOHF. Autoimmune hepatitis was thought to be the cause because of positive results for serum anti-nuclear antibody at 80 times dilution and elevated gamma globulin, but treatment with glucocorticoid did not suppress the progressive hepatic failure. Supportive therapy, including pulse therapy with 1g methylprednisolone for 3 days, ursodesoxycholic acid, branched-chain amino acid, and azathioprine did not resolve the hepatic failure. She was treated by repeated plasmapheresis and plasma absorption for 10 months, and then received the left lobe of her mother's liver. (Her mother's blood type was AB). The patient had been well, being treated with tacrolimus and prednisolone, although the serum titer of anti-blood type B antibody was high just after LRLT and mild liver dysfunction continued for more than 3 years after LRLT. Follow-up biopsy 3 years after LRLT revealed chronic hepatitis and progression to liver cirrhosis. Re transplantation is now under consideration; the patient is now aged 19 years. PMID- 9853570 TI - Multiple focal nodular hyperplasia. AB - Focal nodular hyperplasia (FNH) is a benign lesion of the liver which usually presents with one or two localizations. We report a patient with history of resection of a biliary cyst, and who had been taking oral contraceptives for the past 18 years, who had multiple localizations of FNH (more than 30 lesions). The largest lesion measured 10.5 x 11 x 12cm. The imaging characteristics of our patient were atypical. A central scar could be demonstrated only in the largest lesion, in an eccentric location. In the other lesions, no scar formations could be detected. Furthermore, imaging characteristics suggested that several of the lesions contained fat. This was confirmed by biopsy. The patient had an associated inflammatory syndrome which could not be otherwise explained. The patient was advised to stop taking the oral contraceptives. Follow-up after 2 years showed that the lesions were unchanged; the inflammatory syndrome persisted. Multiple localizations of FNH are very rare. Sometimes they are associated with malformations in other organs (vascular malformations and neoplasia, mostly of the brain). Often they occur as isolated cases, however. Usually their prognosis seems to be good. PMID- 9853571 TI - Three patients with autoimmune cholangiopathy treated with prednisolone. AB - We describe three patients with autoimmune cholangiopathy, i.e., anti mitochondrial antibody-negative and anti-nuclear antibody-positive primary biliary cirrhosis, who were treated with prednisolone. Serum anti-mitochondrial antibody and anti-pyruvate dehydrogenase-E2 component antibody were determined by immunofluorescence of frozen sections and enzyme-linked immunosorbent assay, respectively. Immunoblotting using mitochondria prepared from rat liver was performed to analyze anti-mitochondrial antibody in detail. Serum from one patient reacted with a 48-kilodalton protein, but sera from the other two patients failed to react with the mitochondrial proteins. There was a marked improvement in liver function test results after prednisolone treatment. Before treatment, liver biopsy in all three patients showed histological features of primary biliary cirrhosis with hepatocellular necrosis. Repeat biopsy during treatment showed marked amelioration of hepatocellular damage in all three patients, although bile duct involvement persisted in two patients. These findings suggest that prednisolone is an effective treatment for hepatocellular damage in patients with autoimmune cholangiopathy, but has little impact on the bile duct involvement. PMID- 9853572 TI - Expression of sulfated carbohydrate in Barrett's esophagus: sugar for sure? PMID- 9853573 TI - Gastric ulcer, motility, and trimebutine. PMID- 9853574 TI - New information about gastrointestinal adrenomedullin. PMID- 9853575 TI - Mechanisms of ethanol-induced impairment in asialoglycoprotein receptor expression and function. PMID- 9853576 TI - Vector manometry of the sphincter of Oddi: a new parameter of pancreatobiliary physiology. PMID- 9853577 TI - Zinc deficiency in Crohn's disease. PMID- 9853578 TI - Hepatic failure due to myeloma-associated amyloidosis. PMID- 9853579 TI - Lack of chromosome 15q11-q13 region involvement in a family with Cowden disease/Bannayan-Zonana syndrome. PMID- 9853580 TI - Multiplicity of glucocorticoid action in inhibiting allograft rejection. AB - Glucocorticoids (GCs) are used as immunosuppressive and antiinflammatory agents in organ transplantation and in treating autoimmune diseases and inflammatory disorders. GCs were shown to exert their antiproliferative effects directly through blockade of certain elements of an early membrane-associated signal transduction pathway, modulation of the expression of select adhesion molecules, and by suppression of cytokine synthesis and action. GCs may act indirectly by inducing lipocortin synthesis, which in turn, inhibits arachidonic acid release from membrane-bound stores, and also by inducing transforming growth factor (TGF) beta expression that subsequently blocks cytokine synthesis and T cell activation. Furthermore, by preferentially inhibiting the production of Th1 cytokines, GCs may enhance Th2 cell activity and, hence, precipitate a long lasting state of tolerance through a preferential promotion of a Th2 cytokine secreting profile. In exerting their antiproliferative effects, GCs influence both transcriptional and posttranscriptional events by binding their cytosolic receptor (GR), which subsequently binds the promoter region of cytokine genes on select DNA sites compatible with the GCs responsible elements (GRE) motif. In addition to direct DNA binding, GCs may also directly bind to, and hence antagonize, nuclear factors required for efficient gene expression, thereby markedly reducing transcriptional rate. The pleiotrophy of the GCs action, coupled with the diverse experimental conditions employed in assessing the GCs effects, indicate that GCs may utilize more than one mechanism in inhibiting T cell activation, and warrant careful scrutiny in assigning a mechanism by which GCs exert their antiproliferative effects. PMID- 9853581 TI - Xenogeneic cell therapy: current progress and future developments in porcine cell transplantation. AB - The multitude of distinct cell types present in mature and developing tissues display unique physiologic characteristics. Cellular therapy is a novel technology with the promise of utilizing this diversity to treat a wide range of human degenerative diseases. Intractable diseases, disorders, and injuries are characterized by cell death or aberrant cellular function. Cell transplantation can replace diseased or lost tissue to provide restorative therapy for these conditions. The limited use of cell transplants as a basis for current therapy can, in part, be attributed to the lack of available human cells suitable for transplantation. This has prevented further realization of the promise of cell transplantation as a platform technology. Accordingly, cell-based therapies such as blood transfusions, for which the cells are readily available, are a standard part of current medical practice. Despite numerous attempts to expand primary human cells in tissue culture, current technological limitations of this approach in regard to proliferative capacity and maintenance of the differentiated phenotype has prevented their use for transplantation. Further, use of human stem cells for the derivation of specific cell types for transplantation is an area of future application with great potential, but hurdles remain in regard to deriving and sufficiently expanding these multipotential cells. Thus, it appears that primary cells are at present a superior source for transplantation. This review focuses on pigs as a source of a variety of primary cells to advance cell therapy to the clinic and implement achievement of its full potential. We outline the advantages and disadvantages of xenogeneic cell therapy while underscoring the utility of transplantable porcine cells for the treatment of human disease. PMID- 9853582 TI - Overcoming the histocompatibility barrier: transferrins as carriers and modulators of immunogenic identity. AB - Proteic molecules were found in the bone marrow that were later identified as transferrins. When applied to transplantation of genetically incompatible bone marrow in supralethally irradiated recipient mice, the transferrins obtained from plasma of bone marrow donors promoted engraftment, permanent hemopoietic chimerism, and donor-type immune character. A combination of donor-matched transferrins and antigens was needed for induction of xenogeneic (interspecies) "tolerance" or unresponsiveness to donor antigens in chemically immunosuppressed mice treated with human transferrins and donor leucocytes. This novel and unique property of transferrins may explain the genesis and maintenance of immunogenic identity and allow a reshaping of the immune system. PMID- 9853583 TI - The hNT human neuronal cell line survives and migrates into rat retina. AB - The present studies were undertaken to determine if hNT cells can survive in the vitreous of the eye and migrate into the retina. The hNT neuronal cell line represents a uniform source of human tissue that may be of use in retinal grafts. hNT cells stored in liquid nitrogen were thawed and labeled with the fluorescent dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI). Thirty thousand cells in 1 microL were injected epiretinally in rat. At survival times of 3, 14, 28, or 56 days, retinal sections were examined quantitatively by epifluorescence to reveal DiI-labeled cells. hNT cells survived in the vitreous at all time points without evidence of vascularization. At 3 days, essentially no hNT cells were found in deep retina, and only very few were attached to retina. At days 14, 28, and 56, hNT cells were found to cluster on the vitreal/retinal interface, and in deeper layers. The clusters of hNT cells took on the shape of a funnel at 14 days, and inverted funnel at 28 days, and by 56 days, populated the photoreceptor layer as a stratum. It is possible that hNT cells took on the morphology and function of photoreceptors. These results suggest that hNT cells injected epiretinally survive in the vitreous at least 56 days, migrate to the retinal/vitreous interface, and may migrate through the retina. This system permits the independent and quantitative evaluation of survival and migratory trophic responses. PMID- 9853584 TI - Intraocular transplantation of E1A-immortalized retinal precursor cells. AB - The purpose of this study was to examine the effect of the ocular environment on the survival, tumorigenicity, and phenotypic marker expression of immortalized retinal precursor cells transplanted into immunocompetent adult and neonatal Sprague-Dawley rats. EIA-NR.3, a rat immortalized retinal precursor cell culture, was used as an inexhaustible source of experimental graft material. These cells were prelabeled with the fluorescent marker dil (1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate) and transplanted intravitreally (50,000 cells per microL) into 11 adult and 31 neonatal Sprague-Dawley rat eyes. At 1 mo posttransplant, animals were sacrificed and retinal tissue sections examined histologically for the presence of grafted cells, signs of tumor formation, and retinal phenotypic marker expression. No obvious signs of tumor formation or rejection were seen in a total of 42 eyes in the immunocompetent hosts. Our results indicate that EIA-NR.3 cells survive at least 1 month in vivo, and can migrate from the vitreous into neuroretinal cell layers. Subpopulations of surviving grafted cells were seen to express photoreceptor markers rhodopsin and recoverin comparably between in vitro and in vivo conditions. However, the number of cells immunoreactive for vimentin and E1A decreased significantly under in vivo conditions. This report represents the first experimental intravitreal transplantation of E1A-immortalized retinal precursor cells into adult and neonatal rats. The intraocular location and environment appears to affect phenotypic expression of surviving grafted cells, especially with respect to vimentin and E1A expression. The fact that E1A-NR.3 cells survived intraocularly at least 1 mo without tumor formation suggests that the cells may continue to be useful for further in vivo studies of experimental retinal transplantation, and effects of histological location on retinal cell phenotype and histogenesis in immunocompetent hosts. PMID- 9853585 TI - Effects of surgical anaesthesia on the viability of nigral grafts in the rat striatum. AB - Only a small proportion of dopamine neurons in nigral grafts typically survive transplantation into the adult striatum. Since many anaesthetics reduce blood flow and disturb a variety of brain metabolites, surgical anaesthesia may be one of the factors that compromise graft survival. Conversely, the lowered core body temperature induced by some anaesthetics might promote the survival of grafted cells by slowing their metabolism. In an initial screen, the widely-used surgical anaesthetic, equithesin, was found to reduce core temperature, mean arterial blood pressure, and to increase the partial pressure of oxygen in arterial blood without producing any significant alteration in arterial pH or the partial pressure of carbon dioxide. In the main experiment, rats with unilateral 6 hydroxydopamine lesions of the nigrostriatal bundle received dopamine-rich embryonic nigral grafts injected into the deafferented neostriatum via previously implanted guide cannulae, which allowed comparison to be made of graft survival after transplantation into awake and in re-anaesthetised animals. There were no significant differences between groups in either the functional effects of the grafts to compensate amphetamine-induced rotation, or in the survival and growth of the grafts as measured in post mortem histology. We therefore conclude that anaesthesia per se is not a major contributory factor in the relatively poor survival of dopamine neurons following transplantation into the rat striatum. PMID- 9853586 TI - The development of intracerebral cell-suspension implants is influenced by the grafting medium. AB - The effect of preparing and grafting embryonic striatal and nigral tissue in four different media was evaluated in vitro and in vivo. The proportion of TH-positive and DARPP-32-positive neurons was determined after 2 days in vitro in standard culture medium following preparation in the different media. The effects were more marked for striatal neurons where DARPP-32 expression in tissue prepared in HBSS was poor compared to other media. TH expression was unaffected by the preparation medium. Striatal grafts derived from tissue prepared and grafted in HBSS were smaller, with fewer DARPP-32 cells, compared to other media. Survival of grafts in combined HBSS and DMEM was very poor. Graft volume and TH cell content was enhanced in tissue prepared in DMEM. These results suggest that preparation protocols optimized for one type of embryonic neuronal population do not necessarily transfer to other neuronal populations. PMID- 9853587 TI - Development of approaches to improve the healing following muscle contusion. AB - Muscle injuries are a challenging problem in traumatology, and the most frequent occurrence in sports medicine. Muscle contusions are among the most common muscle injuries. Although this injury is capable of healing, an incomplete functional recovery often occurs, depending on the severity of the blunt trauma. We have developed an animal model of muscle contusion in mice (high energy blunt trauma) and characterized the muscle's ability to heal following this injury using histology and immunohistochemistry to determine the level of muscle regeneration and the development of scar tissue. We have observed a massive muscle regeneration occurring in the first 2 wk postinjury that is subsequently followed by the development of muscle fibrosis. Based on these observations, we propose that the enhancement of muscle growth and regeneration, as well as the prevention of fibrotic development, could be used as approach(es) to improve the healing of muscle injuries. In fact, we have identified three growth factors (bFGF, IGF-1, and NGF) capable of enhancing myoblast proliferation and differentiation in vitro and improving the healing of the injured muscle in vivo. Furthermore, the ability of adenovirus to mediate direct and ex vivo gene transfer of beta-galactosidase into the injured site opens possibilities of delivering an efficient and persistent expression of these growth factors in the injured muscle. These studies should help in the development of strategies to promote efficient muscle healing with complete functional recovery following muscle contusion. PMID- 9853588 TI - The potential contribution of rejection to survival of transplanted human islets. AB - Clinical islet transplantation is potentially the treatment of choice for people with type I diabetes. Rates of insulin independence in islet transplant recipients are disappointingly low, and the relative contribution of the rejection response compared with the loss of islet function is still unclear. We have compared the mixed lymphocyte islet coculture (MLIC) with the mixed lymphocyte acinar cell coculture (MLAC) and the mixed lymphocyte response (MLR) as in vitro models of allograft rejection to MHC and tissue-specific antigens expressed by human islets and acinar cells. The reduced number of MHC class II antigen-positive cells in islets and acinar tissue compared to those in the stimulator lymphocyte population of the MLR, correlated with a reduced proliferative response in the MLIC and MLAC. Enhancement of MHC class II antigen expression by islets using TNFalpha and IFNgamma did not increase their stimulatory capacity in the islet cocultures, which may have been due to a corresponding absence of B7 expression. The lack of T cell proliferation to acinar cells despite cytokine-induced enhancement of MHC class II expression and detectable B7 expression appeared to be due to the inhibitory effect of exocrine enzymes on lymphocyte proliferation. In conclusion, we suggest that a rejection response to islets and acinar tissue is possible due to the accompanying MHC class II-positive cells and that, in this model, islet and acinar-specific antigens do not significantly contribute to that response. Acinar cells may have the potential to stimulate lymphocytes directly, but this was not evident by proliferation in the MLAC. Rejection appears to contribute to the low survival rate of human islet allografts, but it is unlikely that this is the sole explanation, and other factors should be considered. PMID- 9853590 TI - Taking stock of spin science. PMID- 9853589 TI - Long-term amelioration of bilirubin glucuronidation defect in Gunn rats by transplanting genetically modified immortalized autologous hepatocytes. AB - Ex vivo gene therapy, in which hepatocytes are harvested from mutants, retrovirally transduced with a normal gene and transplanted back into the donor, has been used for correction of inherited metabolic defects of liver. Major drawbacks of this method include limited availability of autologous hepatocytes, inefficient retroviral transduction of primary hepatocytes, and the limited number of hepatocytes that can be transplanted safely. To obviate these problems, we transduced primary hepatocytes derived from inbred bilirubin-UDP-glucuronosyl transferase (BUGT)-deficient Gunn rats by infection with a recombinant retrovirus expressing temperature-sensitive mutant SV40 large T antigen (tsT). The immortalized cells were then transduced with a second recombinant retrovirus expressing human B-UGT, and a clone expressing high levels of the enzyme was expanded by culturing at permissive temperature (33 degrees C). At 37 degrees C, tsT antigen was degraded and the cells expressed UGT activity toward bilirubin at a level approximately twice that present in normal rat liver homogenates. For seeding the cells into the liver bed, 1 x 10(7) cells were injected into the spleens of syngeneic Gunn rats five times at 10-day intervals. Excretion of bilirubin glucuronides in bile was demonstrated by HPLC analysis and serum bilirubin levels were reduced by 27 to 52% in 40 days after the first transplantation and remained so throughout the duration of the study (120 days). None of the transplanted Gunn rats or SCID mice transplanted with the immortalized cells developed tumors. PMID- 9853591 TI - Bioinformatics. Gene expression patterns and cancer. PMID- 9853592 TI - Protein-site targeting. Diversification of the drug discovery process. PMID- 9853593 TI - Not quite pharmacogenomics. PMID- 9853594 TI - Industrial postdocs at risk. PMID- 9853595 TI - European biotechnology firms approve of stock market mergers. PMID- 9853596 TI - DPI delivers biotechnology superstore. Discovery Partners International. PMID- 9853597 TI - Restored credibility for sepsis therapeutics? PMID- 9853598 TI - New German government muddies the biotech waters. PMID- 9853599 TI - Naked DNA vaccines come of age. PMID- 9853600 TI - Smaller US firms more aggressive with federal grants. PMID- 9853601 TI - Monsanto unaffected by merger halt? PMID- 9853602 TI - Trimmed down Chiron pitches cancer genomics. PMID- 9853603 TI - Taiwan promotes biotechnology. PMID- 9853604 TI - Sampling the universe of gene expression. PMID- 9853605 TI - Long-lasting gene repair. PMID- 9853606 TI - Genotyping by mass spectrometry takes flight. PMID- 9853607 TI - A singular gene doubles up pest resistance. PMID- 9853608 TI - Modeling the oddities of biology. PMID- 9853609 TI - New antimicrobials--sequence and serendipity in San Diego. PMID- 9853610 TI - Antisense 98: work in progress. PMID- 9853611 TI - The demise of public data on the web? AB - Data rich but cash poor, many free biological databases are on the verge of financial collapse. PMID- 9853612 TI - Out on the farm with DNA vaccines. AB - DNA vaccination is a rapidly developing technology that offers new approaches for the prevention of disease. This technology may permit the production of new vaccines against diseases that have no current vaccine, as well as allowing the development of improved vaccines to replace existing products. We describe how DNA vaccination is being developed for use in commercial animal production, with an emphasis on viral diseases, and discuss the existing hurdles to its development and use. PMID- 9853613 TI - A genome-wide functional assay of signal transduction in living mammalian cells. AB - We describe a genome-wide functional assay for rapid isolation of cell clones and genetic elements responsive to specific stimuli. A promoterless beta-lactamase reporter gene was transfected into a human T-cell line to generate a living library of reporter-tagged clones. When loaded with a cell-permeable fluorogenic substrate, the cell library simultaneously reports the expression of a large number of endogenous genes. Flow cytometry was used to recover individual clones whose reporter-tagged genes were either induced or repressed following T-cell activation. Responsive clones were expanded and analyzed pharmacologically to identify patterns of regulation associated with specific genes. Although demonstrated using T cells, the genomic assay could be applied to map downstream transcriptional consequences for any propagating cell line in response to any stimulus of interest. PMID- 9853614 TI - Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast. AB - We describe a procedure for isolating agonists for mammalian G protein-coupled receptors of unknown function. Human formyl peptide receptor like-1 (FPRL-1) receptor, originally identified as an orphan G protein-coupled receptor related to the formyl peptide receptor (FPR1), was expressed in Saccharomyces cells designed to couple receptor activation to histidine prototrophy. Selection for histidine prototrophs among transformants obtained with a plasmid-based library encoding random peptides identified six different agonists, each of whose production yielded autocrine stimulation of the receptor expressed in yeast. A synthetic version of each peptide promoted activation of FPRL-1 expressed in human embryonic kidney (HEK293) cells, and five of the peptides exhibited significant selectivity for activation of FPRL-1 relative to FPR1. One selective peptide was tested and found to mobilize calcium in isolated human neutrophils. This demonstrates that stimulation of FPRL-1 results in neutrophil activation and suggests that the receptor functions as a component of the inflammatory response. This autocrine selection protocol may be a generally applicable method for providing pharmacological tools to evaluate the physiological roles of the growing number of mammalian orphan G protein-coupled receptors. PMID- 9853615 TI - Selection system for genes encoding nuclear-targeted proteins. AB - Nuclear proteins have essential roles in cell proliferation and differentiation. We have developed a yeast selection system-the nuclear transportation trap (NTT) to identify genes encoding nuclear transport signals. Both unknown and previously identified nuclear localization signals were identified from a human fetal brain cDNA library. The majority (75%) of the unknown proteins examined were exclusively localized to the nucleus in COS-7 cells. We propose that NTT is an efficient method for isolating cDNAs that encode nuclear targeted proteins that can be applied to the retrieval of novel nuclear proteins and to annotate gene function. PMID- 9853616 TI - Stable and inheritable changes in genotype and phenotype of albino melanocytes induced by an RNA-DNA oligonucleotide. AB - Experimental strategies have been developed to correct point mutations using chimeric oligonucleotides composed of RNA and DNA. We used these RNA-DNA oligonucleotides to correct a point mutation in mouse tyrosinase, a key enzyme for melanin synthesis and pigmentation. Melanocytes derived from albino mice contain a homozygous point mutation (TGT-->TCT) in the tyrosinase gene, resulting in an amino acid change from Cys-->Ser. Correction of this point mutation results in the restoration of tyrosinase activity and melanin synthesis, thus changing the pigmentation of the cells. Upon transfection of the RNA-DNA oligonucleotide to albino melanocytes, we detected black-pigmented cells and isolated multiple single clones. All black-pigmented clones exhibited a correction of the point mutation in a single allele of the tyrosinase gene. A full-length tyrosinase was detected by an antityrosinase antibody, and the enzymatic activity was restored in all converted black-pigmented clones. Only degraded fragments were detected in albino cells due to proteolytic cleavage of mutant tyrosinase. The phenotype and genotype of converted black-pigmented clones was stable. These results demonstrate a permanent and stable gene correction by the RNA-DNA oligonucleotide at the level of genomic sequence, protein, and phenotypic change by clonal analysis. PMID- 9853617 TI - High level multiplex genotyping by MALDI-TOF mass spectrometry. AB - A primer extension assay is used to perform highly multiplexed genotyping of single nucleotide polymorphisms (SNPs) present in genomic DNA amplified by a multiplex PCR. The assay uses matrix-assisted laser desorption ionization time-of flight mass spectrometry to accurately measure the masses of short oligonucleotide primers extended by a single dideoxynucleotide. The multiplexed genotyping assays rely on the natural molecular weight differences of DNA bases. By careful analysis of primer composition complementary to the target, or by judicious addition of one or more noncomplementary 5' bases to the genotyping primers, mass spectra of interleaved genotyping products can be generated with no ambiguity in allele assignment. Using a model multiplex PCR system, we demonstrate the ability to perform 12-fold multiplex SNP analysis. PMID- 9853618 TI - Genotyping by mass spectrometric analysis of short DNA fragments. AB - A method has been developed to produce small DNA fragments from PCR products for analysis of defined DNA variations by mass spectrometry. The genomic region to be analyzed is PCR-amplified with primers containing a sequence for the type IIS restriction endonuclease Bpml. Bpml digestion of the resultant PCR products yields fragments as small as seven bases, which are then analyzed by electrospray ionization mass spectrometry. The approach was validated using seven different variants within the APC tumor suppressor gene, in which a perfect correlation was obtained with DNA sequencing. Both the sense and antisense strands were analyzed independently, and several variants can be analyzed simultaneously. These results provide the basis for a generally applicable and highly accurate method that directly queries the mass of variant DNA sequences. PMID- 9853619 TI - Cell surface expression of a human IgG Fc chimera activates macrophages through Fc receptors. AB - Antibody-dependent cell-mediated cytotoxicity plays an important role in the macrophage-mediated destruction of target cells. While the selectivity is based on antibody specificity, the lytic attack is triggered by Fc receptor-mediated respiratory burst. To mimic IgG opsonization, a chimeric antibody-like molecule, containing human IgG1 Fc, was expressed on the surface of mammalian cells. The transmembrane domain of the human transferrin receptor was fused in-frame to the N-terminus of the second and third domains of human IgG1 heavy-chain constant region. This fusion molecule was designed to take advantage of the type II membrane anchor property of the transferrin receptor to express the Fc portion of the molecule in a reverse orientation, such that the Fc portion projected away from the cell surface. This is in contrast to the conventional cell surface IgG, which is anchored by a C-terminal type I transmembrane domain. The cell surface expressed reverse Fc no longer activated complement, but retained Fc receptor binding capability and activated superoxide production by macrophages. This activity was completely blocked by an FcgammaR I-specific monoclonal antibody. PMID- 9853620 TI - A humanized monoclonal antibody produced in transgenic plants for immunoprotection of the vagina against genital herpes. AB - The ability to produce monoclonal antibodies (Mabs) in plants offers the opportunity for the development of an inexpensive method of mucosal immunoprotection against sexually transmitted diseases. To investigate the suitability of plant-expressed Mabs for vaginal preventive applications, we compared a humanized anti-herpes simplex virus 2 (HSV-2) Mab expressed in mammalian cell culture with the same antibody expressed in soybean. We found these Mabs to be similar in their stability in human semen and cervical mucus over 24 h, their ability to diffuse in human cervical mucus, and their efficacy for prevention of vaginal HSV-2 infection in the mouse. PMID- 9853622 TI - Negotiating the maze of biotech "tool patents". AB - Circumventing existing patents may be a shortcut to getting your recombinant protein to market. PMID- 9853621 TI - The tomato Mi-1 gene confers resistance to both root-knot nematodes and potato aphids. AB - Mi-1, a Lycopersicon peruvianum gene conferring resistance to the agricultural pests, root-knot nematodes, and introgressed into tomato, has been cloned using a selective restriction fragment amplification based strategy. Complementation analysis of a susceptible tomato line with a 100 kb cosmid array yielded a single cosmid clone capable of conferring resistance both to the root-knot nematode Meloidogyne incognita and to an unrelated pathogen, the potato aphid Macrosiphum euphorbiae. This resistance was stable. The Mi-1 gene encodes a protein sharing structural features with the nucleotide-binding site leucine-rich repeat containing type of plant resistance genes. PMID- 9853624 TI - Industry trends. Biotechnology in 1998 and beyond. PMID- 9853623 TI - Prediction of antisense oligonucleotide efficacy by in vitro methods. PMID- 9853625 TI - Pharmacy organizations form credentialing council to integrate efforts. PMID- 9853626 TI - Mississippi moves forward with disease-specific credentialing. PMID- 9853627 TI - Pew Commission calls for ongoing competency assessment of health professionals, greater say for public on regulatory boards. PMID- 9853628 TI - Once-daily nonnucleoside reverse-transcriptase inhibitor approved. PMID- 9853629 TI - U.S. health system short on quality, consensus panel says. PMID- 9853630 TI - One fourth of PBM pharmacists' phone calls lead to therapy changes. PMID- 9853631 TI - FDA's Rx for keeping educational programs free of industry bias. PMID- 9853632 TI - Depression in older adults. PMID- 9853633 TI - The Kaiser Permanente/USC Patient Consultation Study: change in use and cost of health care services. AB - The impacts of three alternative models of pharmacist consultation on the use and cost of health care services were studied. Two studies were conducted concurrently in an HMO over two years. In one, 6000 patients were randomly assigned to one of three consultation models; in the other, the three models were implemented in six geographic regions of California (4600 patients). The models were (1) consultation about new or changed prescriptions as mandated by state law (state model), (2) consultation focused on selected high-risk ambulatory care patients (Kaiser Permanente [KP] model), and (3) a control model. The patients were surveyed three times about their health status and satisfaction, and computerized data on health care use and cost were collected. The effect of the consultation models on the use and cost of health care services was examined across five risk groups that were based on drug-use profiles. An additional 37,750 patients (10% of the patients residing in the areawide study sites) were included in a supplemental analysis of the use and cost of health care services. There was no indication in the random-assignment study that pharmacist consultations affected either drug costs or the cost of office visits. Similar results were found in the areawide study, with the exception that the KP model was associated with lower drug costs than the control model. In the 10% sample, the KP model appeared to be associated with lower office visit costs but higher drug costs. Both models were associated with a lower likelihood of a hospital admission and with lower total health care costs for some high-risk patients compared with the control model. Counseling patients about their medications may be unlikely to reduce medication costs or the cost of office visits but may reduce the likelihood of hospital admissions and the overall costs of health care services; a combination of counseling patients at high risk for drug-related problems and counseling all patients about any new or changed prescription should be considered. PMID- 9853634 TI - Continuity of care from acute to ambulatory care setting. AB - Pharmacists' perceptions of a pharmacy-to-pharmacy referral form designed to promote continuity of care for patients being discharged from a hospital were studied. Patients being discharged from a nonteaching, tertiary care hospital to a private residence and having at least one prescription for a new medication were invited to participate in the study. A pharmacy-to-pharmacy referral form was completed on the basis of each patient's medical record and discharge orders. Patients were provided with a copy of the form and told to give it to their community pharmacist with their discharge prescriptions; an additional copy was mailed the same day to the pharmacy. The pharmacist was instructed to telephone the hospital pharmacy on receipt of the form to schedule a telephone interview. A total of 127 patients were enrolled in the study; the average number of discharge medications per patient was 4.8. A total of 120 interviews were conducted with 91 community or ambulatory care pharmacists. In 102 cases (85%), the pharmacists indicated that the referral form provided information that resulted in a benefit to the patient through indication-specific counseling. In 104 cases (87%), pharmacists indicated that the form provided information that had a positive impact on patient-pharmacist interactions, and 75 interviews (62%) indicated that pharmacists were able to incorporate the information into a database to direct future patient care. Community and ambulatory care pharmacists who received information from hospital pharmacists about patients being discharged from the hospital reported that the information aided in patient care. PMID- 9853635 TI - Improved use of time after structural and staffing changes to an ambulatory care pharmacy. PMID- 9853636 TI - Restricting home glucose-monitoring strips in patients taking oral antidiabetic agents. PMID- 9853637 TI - Stability of granisetron hydrochloride in two oral suspensions. PMID- 9853638 TI - ASHP therapeutic position statement on the recognition and treatment of depression in older adults: American Society of Health-System Pharmacists. PMID- 9853639 TI - Report of the ASHP Task Force on Science: American Society of Health-System Pharmacists. PMID- 9853640 TI - Promoting education about complementary or alternative medical therapies. PMID- 9853641 TI - Dosage adjustment of antiretroviral agents in patients with organ dysfunction. PMID- 9853642 TI - Microbial contamination of benzalkonium chloride products. PMID- 9853643 TI - Outsourcing radiopharmaceutical services. PMID- 9853644 TI - Applying classifications of sleep disorders to children with neurologic conditions. AB - Information concerning sleep ontogeny and sleep disorders in children is required by many pediatric specialists. Pediatric neurologists, for instance, frequently are called upon to assist in the evaluation of children with undiagnosed symptoms and signs during sleep, as well as to care for children and adolescents with specific neurologic diseases who also experience sleep disturbances. This review discusses pediatric sleep disturbances with specific reference to sleep in children with neurologic conditions. PMID- 9853645 TI - Fosphenytoin in infants. AB - In comparison with phenytoin preparations, which have a pH value of 11, fosphenytoin, a phosphorylated prodrug of phenytoin, has a pH value of only 8.6, which decreases the risk of cardiovascular and cutaneous side effects. The near neutral pH value of fosphenytoin allows effective intravenous or intramuscular administration. A 1-mg phenytoin equivalent (PE) of fosphenytoin is converted to 1 mg of phenytoin in adults. We describe four infants whose seizures were treated with intravenous fosphenytoin. We had difficulty maintaining therapeutic serum phenytoin levels of 10 to 20 microg/mL on doses of 5 to 8 mgPE/kg/day, and many bolus doses of 5 to 10 mgPE/kg or maintenance doses of more than 10 mgPE/kg/day were given. Despite increased doses in three out of the four patients, a therapeutic serum phenytoin level was not maintained. From our experience, careful and individual dosing of fosphenytoin in this age group can be considered. PMID- 9853646 TI - Electroencephalogram and clinical focalities in juvenile myoclonic epilepsy. AB - Partial seizures and asymmetric abnormalities seen on electroencephalogram (EEG) are infrequent in juvenile myoclonic epilepsy, but when present, can lead to a misdiagnosis of partial seizures. We report four patients with juvenile myoclonic epilepsy who had generalized spike or polyspike and wave discharges on EEG in addition to clinical and EEG evidence of focality. The clinical course and response to therapy was similar to that in typical juvenile myoclonic epilepsy. PMID- 9853647 TI - Serum carnitine levels in epileptic children before and during treatment with valproic acid, carbamazepine, and phenobarbital. AB - Serum levels of free, acyl, and total carnitine were determined in 32 patients with seizures, before and after 3, 6, and 12 months of treatment with valproic acid (17 patients), carbamazepine (10 patients), or phenobarbital (5 patients). In all three treated groups, both free and total carnitine levels showed a significant decline with respect to pretreatment levels. This decline was most marked and most consistent in patients treated with valproic acid. In 35% of the patients in this group, carnitine deficiency (ie, total carnitine < 30 micromol/L) was observed by month 12. In none of the three groups were serum carnitine levels significantly correlated with the serum concentration of the drug. These findings suggest a need to monitor serum carnitine levels in children treated with any of these drugs. PMID- 9853648 TI - Developmental changes of cortical and cerebellar motor control: a clinical positron emission tomography study with children and adults. AB - Functional neuroimaging data regarding the development of motor organization in normal children and adolescents are virtually unavailable because of ethical concerns. As an alternative approach, we studied child and adult lesion patients, focusing on movement of the hand ipsilateral to the lesion and on brain activations in the contralesional hemisphere. [15O]-water positron emission tomography was performed during rest and sequential finger-thumb tapping in 10 children (aged 6 to 14 years) and 15 adults (aged 18 to 74 years) with unilateral lesion. We expected more distinct activation/deactivation patterns during movement in adults than in children. While there were no group differences in activation of primary and secondary motor cortices, deactivations in nonmotor cortex were significantly more pronounced in adults than in children. This indirectly supports our hypothesis of developmental focalization of cerebral motor control. Activations in the cerebellum and vermis were significantly stronger in the adults than in the children, possibly reflecting normal developmental patterns. PMID- 9853649 TI - Beta-interferon plus inosiplex in the treatment of subacute sclerosing panencephalitis. AB - We treated seven patients with subacute sclerosing panencephalitis with beta interferon and oral inosiplex for 2 to 15 months. Stabilization or improvement was observed in three patients. The effect of treatment was equivocal in two other patients who became stable. The disease continued its progression in the remaining two patients who died. Treatment shorter than 2 months was not effective. Changes in electroencephalograms (EEG), magnetic resonance images (MRI), or cerebrospinal fluid measles antibody levels did not have a close correlation with clinical course. These results suggest that beta-interferon might be efficient in some patients with subacute sclerosing panencephalitis and justify its trial in larger studies with longer follow-up. PMID- 9853650 TI - Citation and impact of the Journal of Child Neurology. PMID- 9853651 TI - Chronic meningoencephalitis due to cytomegalovirus infection in a nonimmunocompromised patient: an autopsy case. PMID- 9853652 TI - A large-scale mitochondrial DNA deletion causing progressive ataxia. PMID- 9853653 TI - Vein of Galen malformation with diencephalic syndrome: a clinical pathologic report. PMID- 9853654 TI - Phenytoin-induced dermatomyositis: case report and literature review. PMID- 9853655 TI - Vertical gaze palsy induced by midbrain lesions and its structural imaging. AB - We experienced four cases of vertical gaze palsy induced by midbrain lesions. Lesions commonly covered the rostral midbrain, including the rostral interstitial nucleus, dorsomedial to the red nucleus. Two of the four cases resulted from vascular insult, in which a single, unpaired perforator is supposed to innervate the rostral midbrain and medial thalamus bilaterally. One case showed vertical gaze palsy accompanied by bilateral ptosis. The findings agree with recent experimental evidence that a neural substrate in eyelid control lies in the supraoculomotor area immediately dorsal to the oculomotor nucleus. The remaining two cases, a brain hemorrhage and an inflammatory tumor, showed unilateral lesions of the rostral midbrain. In these cases, vertical gazes were not abolished, but were limited in an incomplete way. This may be explained by partial damages of the descending fibers, some of which decussate through the posterior commissure before it reaches the oculomotor nucleus. Thus, clinical signs and symptoms were clarified based on anatomical and physiological points of view. PMID- 9853656 TI - The effect of respiratory mode on human middle ear pressure. AB - The purpose of this study was to examine the effect of respiratory mode on the human middle ear pressure. The partial pressure of carbon dioxide in the venous blood (PvCO2) and the middle ear pressure were increased by hypoventilation, and decreased by hyperventilation. The partial pressure of oxygen in the venous blood showed little change. It was considered that CO2 diffused into the middle ear cleft according to the partial pressure gradient when the PvCO2 was elevated by hypoventilation, resulting in increase of the middle ear pressure, whereas CO2 diffused into the blood from the middle ear cleft when the PvCO2 was reduced by hyperventilation, resulting in decrease of the middle ear pressure. These findings suggest that bidirectional gas exchange via the middle ear mucosa functions actively in the human middle ear and plays an essential role in the regulation of the middle ear pressure. PMID- 9853657 TI - Short-latency EMG potentials elicited by head taps in sternocleidomastoid muscles: a study on normal human subjects and patients with central or vestibular lesions. AB - The aim of the present study was to determine whether a tap on the forehead evokes a vestibulospinal reflex in sternocleidomastoid muscles in normal human subjects and patients with central or vestibular lesions. While first positive negative short-latency EMG potentials with a mean positive potential peak of about 12 ms and a mean negative potential peak of about 17 ms were found in the bilateral sternocleidomastoid muscles in normal subjects, they disappeared or were attenuated in patients with central or peripheral vestibular lesions. PMID- 9853658 TI - Timing for removal of tympanic ventilation tube in children. AB - The medical records of 220 ears of 137 pediatric patients (85 male and 52 female) in which three kinds of ventilation tubes were inserted for treating otitis media with effusion (OME) were reviewed. The tubes selected were the Shepard grommet (75 ears), Goode-T (39 ears), and Paparella type II tube (106 ears). The criteria for tube placement were as follows: (1) continuous conductive hearing loss with over 25 dB air-bone gap, (2) resistance to conservative therapy for over 6 months, and (3) retracted and glue-colored tympanic membrane with type B tympanogram. The tubes that remained in place for over 18-24 months were removed intentionally in combination with a freshening of the perforation edge and tape patch technique using Steri-Strip tape (3M) for preventing permanent eardrum perforation, because the incidence of persistent perforation became higher after long-term intubation. Shepard grommets tended to be extruded earlier, while Paparella type II tubes tended to stay longer. The OME recurrence rate decreased 12 months or more after tubal insertion. There was a tendency for the recurrence rate to decrease the longer the tube stayed in the eardrum. The number of recurrences decreased when the patient's age at the tube removal or extrusion was 7-8 years old. Adenoidectomy did not influence the recurrence rate of OME. Although the Goode-T and Paparella tube II tubes showed high perforation rates, the perforation rate after extrusion or removal of the tube was decreased by the use of the tape patch technique in combination with a freshening of the perforation edge. From these findings, it was concluded that the appropriate intubation period for the treatment of OME in children is over 12 months with the use of a long-term tube, and that if the patient's age at the time of tube insertion was below 6 years, it might be better that the removal of the tube is postponed until the patient is 8 years of age. PMID- 9853659 TI - Hearing parameters in noise exposed industrial workers. AB - This paper presents the results of a study carried out in a group of noise exposed workers in a hydro-electric power plant. Thus, the main focus of the study is on 130 industrial workers who were exposed to high level of noise. The control group was consisted of 33 subjects with normal hearing. Hearing and acoustic reflex thresholds were obtained from all subjects and the results from age-matched subgroups were compared. The sensorineural hearing loss which were detected in 71 workers were bilateral, symmetrical and affected mainly frequencies of 4-6 kHz. In essence, the hearing losses were developed within the first 10 years of noise exposure and associated with slight progress in the following years. When acoustic reflex thresholds derived from the study and control groups were compared, statistically significant difference was determined only for the thresholds obtained at 4 kHz (p < 0.0005). PMID- 9853660 TI - Response of EP and cochlear blood flow to angiotensin II during hypoxic condition. AB - To understand the importance of oxygen transport to the inner ear tissue, we studied, using guinea pigs, the relationship between endocochlear DC potential (EP) and cochlear blood flow (CoBF) under hypoxic conditions. 33 guinea pigs weighing 200-400 g were used. EP and CoBF were recorded from the basal turn of the cochlea. Hypoxia (10 min) was induced by inspiring a mixture gas (2-10% oxygen in nitrogen). By inducing hypoxia, the EP declined to a intermediate level of the positive range in a few minutes. In the midpoint of 10-min loading of hypoxia, Angiotensin II (1 microg/kg, 1 ml/kg) was infused in 60 s, resulting in raising of systemic blood pressure (BP). Responding to this, CoBF and EP showed generally transient elevations (deltaCoBF, deltaEP) in accordance with BP change. Obtained data were analyzed for the value of deltaEP/deltaCoBF, which means the sensitivity of EP to CoBF change. Distribution of the value of deltaEP/deltaCoBF was found to be in a correlation with the EP level just before Angiotensin II infusion (EPi). That is, the sensitivity of EP to CoBF change became greater as a intermediate level of EP (EPi) was lowered in the range from 70 to 100% of EPi. Among several factors, we thought the hyperactivity of glycolysis in the stria vascularis may occur in such a mild hypoxemic condition. In addition to this, it could be presumable that cyclic AMP in the stria vascularis may increase by the sympathetic stimulation during hypoxemia, and increased cyclic AMP may produce more EP with a given O2 consumption. PMID- 9853661 TI - Detection of herpes simplex virus-1 by nested PCR. An experimental model. AB - Nested polymerase chain reaction (nested PCR) was performed using a reaction mix batch-prepared and kept frozen in single reaction tubes at -20 degrees C until use. Twenty-one New Zealand white rabbits were infected with herpes simplex virus type 1 (HSV-1). Eleven animals were killed on day seven and the other ten were sacrificed on day 21. Viral culture and nested PCR was used to determine the presence of HSV-1 in samples from the tongue, HSV-1 was detected in 90.47% of the animals; in 84.21% by nested PCR and in 52.63% by culture. Nested PCR assay had greater sensitivity than culture in animals sacrificed on day seven with significative difference (p < 0.05). Higher sensitivity and faster results were obtained with this method, so we found it reliable and useful in the setting of a clinical laboratory dealing with diagnosis of herpes virus infections. PMID- 9853662 TI - Squamous cell carcinoma of the nasal septum with Wegener's granulomatosis treated with cyclophosphamide and corticosteroids. AB - Well differentiated squamous cell carcinoma of the nasal septum developed in a 55 year old man with Wegener's granulomatosis. It is suggested that the malignancy was induced by immunosuppressive state from an increased and prolonged use of cyclophosphamide and corticosteroids. Although the efficacy of the therapeutic concept using cyclophosphamide and corticosteroids is well established, there have been some few reports that cyclophosphamide could be implicated in the genesis of malignancies. The pathophysiology of Wegener's granulomatosis should be better understood, and effective and less toxic alternative protocol should be established. PMID- 9853663 TI - The significance of CT scan or MRI in the evaluation of salivary gland tumors. AB - Imaging modalities such as CT scan or MRI are frequently employed for the diagnosis of neoplastic lesions in the salivary glands. To evaluate the efficacy of the CT scan and the MRI in differentiating malignant neoplasm from benign lesions, 120 CT scans and 31 MRIs were retrospectively analyzed from 147 patients with salivary gland masses. All images were analyzed focusing on the presence of several relevant features. The pathologic results were matched with radiological features and also tabulated with radiological assessment. For the CT scans, the contour and margin of the lesion and tissue plane obliteration were found to be statistically significant indicators for malignant neoplasms. Among 69 CT scans interpreted as 'benign' by a radiologist, five cases (7%) were histologically diagnosed as 'malignant'. On the other hand, 20 out of 51 CT scans (39%) were misinterpreted as 'malignant'. For MRI, two out of 14 cases (14%) were radiologically misdiagnosed as 'benign' and six out of 17 patients (35%) as 'malignant'. In conclusion, whereas both the CT and MRI showed a similar level of accuracy in evaluation of salivary gland tumors, they showed a considerable tendency of misdiagnosis, especially by interpreting benign tumors as 'malignant'. PMID- 9853664 TI - Retropharyngeal abscess: recent trends. AB - Retropharyngeal abscess (RPA) is relatively rare today. A study of 17 cases of RPA treated at our hospital in the past 10 years showed a shift in the disease from children below 6 years of age (41%) to older children and adults (58%). Upper respiratory tract infection (URTI) was found to be the commonest (52%) aetiological predisposing factor in all age groups. Other aetiological factors were septicaemia (11%) in children below the age of 6 years and trauma due to foreign body (35%) in the older children and adult age groups. Klebsiella, Staphylococcus and Streptococcus were the commonest species of microorganisms grown from pus. The changing clinical trends, microbiology, choice of antibiotics, usefulness of radiology, and complications of this potentially fatal illness are discussed. PMID- 9853665 TI - Long term follow-up study of laryngeal precancer. AB - The goal of this study was to determine subsequent malignancy on a discrete group of precancerous laryngeal lesions, and to assess the mortality. In a series of 227 patients, average age 51.8 years (ranging from 13 to 80 years). The changes were followed-up for 12.3 years (minimum of 5 years and maximum of 40 years). 58% are living without any sign of premalignant laryngeal mucosal disease, 13% with controlled precancer, and 3% in remission after surgery for carcinoma. 11% died (9% due to cause unrelated to the cancer) and 15% were lost for follow-up. 17% of the group with mucosal hyper- or metaplasia progressed to mild dysplasia, but none progressed to carcinoma. Reinke's oedema recurred in 4%, no malignancy was observed. Carcinoma developed in 16% of laryngeal papilloma (8% in situ and 8% invasive). 15% of mild dysplasia progressed in severity, but none transformed to malignancy. Moderate dysplasia progressed to severe dysplasia in 12%, carcinoma in situ in 4%. Of cases with severe dysplasia 13% developed in situ carcinoma while 43% progressed to invasive cancer. In the whole series progression to severe grade was seen in 7.1%, and malignant transformation in 4.4%. Three patients (1.3%) died due to subsequent carcinoma. Our results agree with some authors; but the majority of them reports higher incidence of malignant transformation. Invasive carcinoma was diagnosed in the follow-up in seven patients. Those represent only 3% of all laryngeal carcinomas diagnosed in our department in the same period of time. Based on the data we have evaluated the intensity of follow-up in patients with hyperplasia, metaplasia, keratosis and mild dysplasia. PMID- 9853666 TI - Chemoprevention with interferon alfa and 13-cis retinoic acid in the adjunctive treatment of head and neck cancer. AB - BACKGROUND: Retinoids have been increasingly used since the mid-1960s for the treatment of leukoplakia and dysplasia of the head and neck. Studies of their use in the treatment of carcinomas of the head and neck, usually in combination with interferons, have also been published in recent years. METHODS: 30 patients (25 men, five women) with UICC stage IV were given adjunctive treatment with a combination of 3 x 3 million IU of interferon alfa SC weekly and 0.5 mg/kg body weight of 13-cis retinoic acid PO daily for a maximum duration of 6 months. The therapeutic benefits and side effects are reported here. RESULTS: Therapy was completed as scheduled in 16 out of 30 patients. Reasons for stopping treatment: progressive disease (ten patients), side effects (four patients). 18 patients were tumor-free following treatment. 16 patients displayed a complete response one year after completion of adjunctive treatment. Retinoic-associated side effects observed included xerostomia (90%), dysphagia (67%), weight loss (50%), flush (50%) and cachexia (7%). Interferon-associated side effects included pyrexia and moderate hematological changes. CONCLUSION: Adjunctive combination treatment with interferon alfa and 13-cis retinoic acid appears to be beneficial to patients with head and neck cancer. The side effects are moderate. PMID- 9853667 TI - Malignancies of human thyroid tumors and dynamic magnetic resonance imaging (MRI). AB - Time intensity curves for gadolinium-diethylene triaminepentacetic acid (Gd-DTPA) enhanced magnetic resonance imaging (MRI), namely dynamic MRI, were determined for thyroid diseases and compared with findings of histopathologic examination. Time intensity curves for solid lesions were determined, excluding cases with secondary changes such as calcification, hemorrhage, necrosis and fibrosis. Three different patterns of time intensity curves were observed: rapid washout, delayed washout and no change. In our previous study, malignant grades of thyroid tumors were estimated immunohistochemically by epidermal growth factor receptor (EGFR) antibody. In most of malignant diseases and a few benign diseases that had marked cell proliferative activity with staining EGFR strongly, the time intensity curve displayed a delayed washout pattern, in which intensity was above 1/2-maximal value within 10 min after injection Gd-DTPA. Almost all benign diseases and a few well differentiated carcinomas displayed a rapid washout pastern, in which intensity was decreased to lower than 1/2 of peak grade within 10 min following injection and showed staining EGFR weakly. Benign diseases showing no change of time intensity curve, did not almost show aEGFR positive cell. These findings suggested that the time intensity curve obtained from dynamic MRI might indicate differentiated grades and cell proliferating activity of thyroid tumors. PMID- 9853669 TI - Embryonal rhabdomyosarcoma of the nose in an elderly patient, case report and review of the literature. AB - Embryonal rhabdomyosarcoma of the head and neck is very rare in adults. We report on an embryonal rhabdomyosarcoma of the nose, occuring in a 74-year-old female patient presenting with nasal congestion as the only clinical symptom. Suspected to be an amelanotic melanoma, further biopsies and histological examinations provided the correct diagnosis. She first underwent a course of chemotherapy which led to a partial shrinkage of the tumor mass which was followed by a complete resection of the residual tumor mass. Biologic behaviour of this tumor must be fully appreciated as treatment is interdisciplinary. Rhabdomyosarcoma should be considered in the differential diagnosis of tumors of the nose and paranasal sinuses not only in children, but also in adults. PMID- 9853668 TI - Antigen retrieval immunohistochemistry used for routinely processed celloidin embedded human temporal bone sections: standardization and development. AB - The use of immunohistochemistry (IHC) in routinely processed celloidin-embedded human temporal bone section has created a fruitful field in understanding the pathogenesis and pathophysiology of the human inner ear at a molecular level since the early 1990s when the antigen retrieval (AR) technique was developed. This review article focuses on several critical technical issues of AR technique based predominantly on our experiences and suggestions concerning further development and standardization of AR-IHC for IHC study of human temporal bone section, as well as other tissues embedded in celloidin. Examples of using AR-IHC in human temporal bone sections collected include our unpublished data in order to indicate the potential utility of this novel method. Suggestions of further development of AR techniques are proposed for references and comments. PMID- 9853670 TI - Acinic cell carcinoma of maxillary sinus. AB - Acinic cell carcinoma is a tumor that occurs most commonly in the parotid glands. We have experienced a rare case of acinic cell carcinoma of the left maxillary sinus. The patient was a 71-year-old female. She noticed her left nasal congestion and epiphora from left eye in April 1994. She had a history of left maxillary sinus surgery in February, 1972. Antrotomy was performed and histopathological examination of specimen from the maxillary sinus proved acinic cell carcinoma. The tumor was considered to be the recurrence 22 years after initial surgery, because it coincided with the surgical specimen of previous surgery. Radical resection of maxilla was performed for complete resection of tumor with palate reconstruction by left scapula and latissimus dorsi myocutaneous flap. No recurrence is observed for more than 4 years after surgery. Although this tumor is of low grade malignancy, complete resection with adequate surgical margin is advisable due to high incidence of this tumor to recur after long periods of time. PMID- 9853671 TI - Endodermal sinus (yolk sac) tumor of oral cavity originating from gingiva. AB - Endodermal sinus tumor is a malignant germ cell tumor which usually arises in gonads. Extragonadal endodermal sinus tumors in the head and neck are very rare. Here a gingival endodermal sinus tumor is reported. The lesion demonstrated typical microscopic features of the endodermal sinus tumors of gonads. The tumor cells showed alpha-fetoprotein immunoreactivity in immunohistochemical evaluation. The serum alpha-fetoprotein level was high. This is the first gingival case in the related literature. PMID- 9853672 TI - Analysis of essential histidine residues of maize branching enzymes by chemical modification and site-directed mutagenesis. AB - Incubation of maize branching enzyme, mBEI and mBEII, with 100 microM diethylpyrocarbonate (DEPC) rapidly inactivated the enzymes. Treatment of the DEPC-inactivated enzymes with 100500 mM hydroxylamine restored the enzyme activities. Spectroscopic data indicated that the inactivation of BE with DEPC was the result of histidine modification. The addition of the substrate amylose or amylopectin retarded the enzyme inactivation by DEPC, suggesting that the histidine residues are important for substrate binding. In maize BEII, conserved histidine residues are in catalytic regions 1 (His320) and 4 (His508). His320 and His508 were individually replaced by Ala via site-directed mutagenesis to probe their role in catalysis. Expression of these mutants in E. coli showed a significant decrease of the activity and the mutant enzymes had Km values 10 times higher than the wild type. Therefore, residues His320 and His508 do play an important role in substrate binding. PMID- 9853673 TI - Structural dissection of the DNA-binding domain of the yeast transcriptional activator GAL4 reveals an alpha-helical region responsible for dimerization. AB - Limited proteolysis of the DNA-binding domain (residues 1-147) of the yeast transcriptional activator GAL4 has been used to define more precisely the subdomain structure required for DNA binding and dimerization. Two regions of the protein were found to be resistant to proteolysis: the cysteine-rich, zinc binding region (residues 6-43) and a hydrophobic sequence between residues 52 and 97. Carboxy-terminal deletion fragments of the DNA-binding domain were generated and assayed by DNase 1 footprinting. This showed that the affinity of DNA binding depends on the sequence between residues 65 and 94. Structural comparisons by UV circular dichroism (CD) were made and the difference CD spectra indicate that strong alpha-helical content is found specifically in the region between residues 65 and 94, which previous studies have shown to enable dimerization and in this study the formation of a stable protein-DNA complex. PMID- 9853674 TI - Two dual-specific (anti-IgG and anti-dsDNA) monoclonal autoantibodies derived from the NZB/NZW F1 recognize an epitope in the hinge region. AB - The anti-IgG properties of two dual-specific (anti-dsDNA and anti-IgG) monoclonal NZB/NZW F1-derived autoantibodies, BV 17-45 and BV 16-13, were studied to resolve the location and possible commonality of the IgG epitope. To determine if BV 17 45 and BV 16-13 recognized the same IgG epitope, the relative temperature sensitivity of the conformational IgG epitopes were evaluated using the conformational sensitive immunoassay. Comparison of the temperature sensitivity of the conformational immunoglobulin epitopes over a temperature range of 25-100 degrees C suggested that the epitope recognized by BV 17-45 was the same as the IgG epitope recognized by BV 16-13. Further studies with papain- and pepsin generated F(ab')2, Fab, and Fc fragments of BV 17-45 and BV 16-13 revealed that the dual-specific autoantibodies BV 17-45 and BV 16-13 both bound an epitope in the hinge region of the IgG molecule. The potential correlation between these studies and the pathogenic nature of dual-specific autoantibodies is discussed. PMID- 9853675 TI - Artificial neural network method for predicting HIV protease cleavage sites in protein. AB - Knowledge of the polyprotein cleavage sites by HIV protease will refine our understanding of its specificity, and the information thus acquired will be useful for designing specific and efficient HIV protease inhibitors. The search for inhibitors of HIV protease will be greatly expedited if one can find an accurate, robust, and rapid method for predicting the cleavage sites in proteins by HIV protease. In this paper, Kohonen's self-organization model, which uses typical artificial neural networks, is applied to predict the cleavability of oligopeptides by proteases with multiple and extended specificity subsites. We selected HIV-1 protease as the subject of study. We chose 299 oligopeptides for the training set, and another 63 oligopeptides for the test set. Because of its high rate of correct prediction (58/63 = 92.06%) and stronger fault-tolerant ability, the neural network method should be a useful technique for finding effective inhibitors of HIV protease, which is one of the targets in designing potential drugs against AIDS. The principle of the artificial neural network method can also be applied to analyzing the specificity of any multisubsite enzyme. PMID- 9853676 TI - Influence of chemical modification of cysteine and histidine side chains upon subunit reassembly of alpha crystallin. AB - Alpha crystallin, the important multimeric structural protein of mammalian eye lens, is an assembly composed of 30 alpha-A and 10 alpha-B subunits. The influence of either partial or complete chemical modification of two important amino acid side chains, cysteine and histidine, upon the integrity of native alpha crystallin assembly and also upon the mode of subunit reassembly has been investigated. It has been found that chemical modification of surface-exposed cysteine and histidine side chains does not affect the subunit-subunit interactions stabilizing the native aggregate. Cysteine modifications, either partial or complete, unlike histidine modifications, do not seem to affect the backbone conformation of the subunits refolded after denaturation. Both cysteine and histidine modifications, however, affect the packing of the refolded structural elements forming the tertiary structure of the subunits and also the mode of oligomeric reorganization. The most striking effect of histidine modification is the considerable increase in size of the aggregates upon reassociation of the modified subunits. The chaperone activity, however, has been found to remain almost unaffected in spite of these chemical modifications. PMID- 9853677 TI - Primary structure of apolipophorin-III from the greater wax moth, Galleria mellonella. AB - The complete amino acid sequence of apolipophorin-III (apoLp-III), a lipid binding hemolymph protein from the greater wax moth, Galleria mellonella, was determined by protein sequencing. The mature protein consists of 163 amino acid residues forming a protein of 18,075.5 Da. Its sequence is similar to apoLp-III from other Lepidopteran species, but remarkably different from the apoLp-IIIs of insects from other orders. As shown by mass spectrometric analysis, the protein carries no modifications. Thus, all of its known physiological functions, including its recently discovered immune response-stimulating activity, must reside in the protein itself. PMID- 9853678 TI - Inhibition of human cytochrome P450 1A2 by flavones: a molecular modeling study. AB - Cytochrome P450 1A2 metabolizes a number of important drugs, procarcinogens, and endogenous compounds. Several flavones, a class of phytochemicals consumed in the human diet, have been shown to differentially inhibit human P450 1A2-mediated methoxyresorufin demethylase. A molecular model of this P450 was constructed in order to elucidate the molecular basis of the P450-flavone interaction. Flavone and its 3,5,7-trihydroxy and 3,5,7-trimethoxy derivatives were docked into the active site to assess their mode of binding. The site is hydrophobic and includes several residues that hydrogen bond with substituents on the flavone nucleus. The binding interactions of these flavones in the modeled active side are consistent with their relative inhibitory potentials, namely 3,5,7-trihydroxylflavone > flavone > 3,5,7-trimethoxylflavone, toward P450 1A2-mediated methoxyresorufin demethylation. PMID- 9853679 TI - The cDNA-derived amino acid sequence of indoleamine dioxygenase like-myoglobin from the gastropod mollusc Omphalius pfeifferi. AB - Myoglobin was isolated from the radular muscle of the archaegastropod mollusc Omphalius pfeifferi (Trochidae). The molecular mass was estimated by SDS-PAGE to be about 40 kDa, 2.5 times larger than that of usual myoglobin. The cDNA for Omphalius myoglobin was amplified by polymerase chain reaction, and the cDNA derived amino acid sequence of 375 residues was determined, of which 73 residues were identified directly by the chemical sequencing of internal peptides. The amino acid sequence of Omphalius myoglobin showed no significant homology with any other usual 16-kDa globins, but showed 84% and 36% identities with indoleamine dioxygenase-like myoglobins from Battilus (Turbinidae) and Sulculus (Haliotiidae), respectively. It also shows significant homology (26% identity) with human indoleamine 2,3-dioxygenase, a tryptophan-degrading enzyme containing heme. The distribution of indoleamine dioxygenase-like myoglobins suggests that they must have arisen exclusively along the specified lineage including the three families Haliotiidae, Turbinidae, and Trochidae of Archaegastropoda in molluscan evolution. PMID- 9853680 TI - A bovine dander allergen, comparative modeling, and similarities and differences in folding with related proteins. AB - The most important allergenic protein in cow dander and urine is Bos d 2. It is proposed to belong to the family of lipocalins, which are proteins capable of binding small hydrophobic molecules. The allergenic properties of Bos d 2 indicate an interaction between the accessible regions of the native protein and IgE. In this work, a three-dimensional model was created for Bos d 2 by comparative modeling, and features characteristic of outlier lipocalins were observed. The protruding regions of the surface were characterized and used in predicting the possible B-cell epitopes. There is a pocket inside the core and its size is appropriate for small molecules. The model shows a hydrophilic amino acid side chain of glutamic acid 115 on the inner surface of the hole and a phenylalanine as the "gatekeeper" instead of tyrosine, which is common in experimentally modeled lipocalins. PMID- 9853681 TI - Podophyllotoxin and nocodazole counter the effect of IKP104 on tubulin decay. AB - Tubulin, the subunit protein of microtubules, undergoes a time-dependent loss of functional properties known as decay. We have previously shown that the drug 2-(4 fluorophenyl)- -(2-chloro-3,5-dimethoxyphenyl)-3-methyl-6-phenyl-4(1H)-pyridinone (IKP104) accelerates decay, but that in the presence of colchicine, IKP104 becomes a stabilizer of tubulin. To see if this is due to conformational effects specific to colchicine or simply to occupancy at the colchicine site, we examined the effects of nocodazole and podophyllotoxin, two well-known competitive inhibitors of colchicine for binding to tubulin, on IKP104's acceleration of decay. We found that podophyllotoxin abolished IKP104's accelerating effect and, like colchicine, turned it into a stabilizer of tubulin. Nocodazole's effects were similar to those of podophyllotoxin and colchicine, in that it abolished IKP104-induced enhancement of decay; however, in the presence of nocodazole, IKP104 caused little or no stabilization of tubulin. Since colchicine, nocodazole, and podophyllotoxin have very different interactions with tubulin, but all inhibit the IKP104-induced enhancement of decay, our findings suggest that this inhibition arises from occupancy of the colchicine site rather than from a direct conformational effect of these two drugs. PMID- 9853682 TI - Chemistry of the "molecular trap" of protease-catalyzed splicing reaction of complementary segments of alpha-subunit of hemoglobin A. AB - The complementary fragments of human Hb alpha, alpha1-30, and alpha31-141 are spliced together by V8 protease in the presence of 30% n-propanol to generate the full-length molecule (Hb alpha-semisynthetic reaction). Unlike the other protease catalyzed protein/peptide splicing reactions of fragment complementing systems, the enzymic condensation of nonassociating segments of Hb alpha is facilitated by the organic cosolvent induced alpha-helical conformation of product acting as the "molecular trap" of the splicing reaction. The segments alpha24-30 and alpha31-40 are the shortest complementary segments that can be spliced by V8 protease. In the present study, the chemistry of the contiguous segment (product) alpha24-40 has been manipulated by engineering the amino acid replacements to the positions alpha27 and alpha31 to delineate the structural basis of the molecular trap. The location of Glu27 and Arg31 residues in the contiguous segment alpha24-40 (as well as in other larger segments) is ideal to generate (i, i + 4) side-chain carboxylate-guanidino interaction in its alpha-helical conformation. The amino acid residue replacement studies have confirmed that the side chains at alpha27 and alpha31 facilitate the semisynthetic reaction. The relative influence of the substitute at these sites on the splicing reaction depends on the chemical nature of the side chain and the location. The gamma-carboxylate guanidino side-chain interaction appears to contribute up to a maximum of 85% of the thermodynamic stability of the molecular trap. The studies also demonstrate that the thermodynamic stability of the molecular trap is determined by two interdependent conformational aspects of the peptide. One is an amino acid-sequence-specific event that facilitates the induction of an alpha-helical conformation to the contiguous segment in the presence of organic cosolvent that imparts some amount of protease resistance to Glu30-Arg31 peptide bond. The second structural aspect is a site-specific event, an i, i + 4 side-chain interaction in the alpha-helical conformation of the peptide which imparts an additional thermodynamic stability to the molecular trap. The results suggest that conformationally driven "molecular traps" of protease-mediated ligation reactions of peptides could be designed into products to facilitate the modular assembly of peptides/proteins. PMID- 9853683 TI - Conformational change of human lens insoluble alpha-crystallin. AB - Human lens alpha-crystallin becomes progressively insoluble with age and is the major crystallin component in the water-insoluble (WI) fraction. The mechanism that causes the originally water-soluble (WS) alpha-crystallin to become insoluble is unknown. A conformational change by chemical modification may be the cause, but the nature of insolubility renders it impossible to study protein conformation in the WI fraction by most spectroscopic measurements. In the present study, alpha-crystallin in the WI fraction was extracted by urea and reconstituted to a folded protein by dialysis. The refolded urea-soluble (US) alpha-crystallin was compared with WS alpha-crystallin. The US alpha-crystallin has a greater amount of polymeric species, but fewer degraded subunits than the WS alpha-crystallin as shown by SDS-PAGE and Western blot. Circular dichroism (CD) measurements indicate that they have the same secondary structure but a different tertiary structure, possibly a partial unfolding in the US alpha crystallin. This is supported by fluorescence measurements: Trp residues are more exposed and protein has a more-hydrophobic surface in the US than in the WS alpha crystallin. Blue fluorescence further indicates that the US alpha-crystallin has a greater amount of pigment than the WS alpha-crystallin. Together, these results indicate that the US alpha-crystallin is a chemically and conformationally modified protein. PMID- 9853684 TI - Distinct and overlapping binding sites for IKP104 and vinblastine on tubulin. AB - IKP104 is one of a group of tubulin-binding drugs whose interaction with tubulin suggests that it may bind to the protein at or close to the region where vinblastine binds. By itself IKP104 is a potent enhancer of tubulin decay as evidenced by the fact that it induces the exposure of the sulfhydryl groups and hydrophobic areas on tubulin. In this respect, IKP104 differs from vinblastine and other drugs such as phomopsin A, dolastatin 10, rhizoxin, and maytansine which are competitive or noncompetitive inhibitors of vinblastine binding. In contrast, however, in the presence of colchicine, IKP104 behaves differently and strongly stabilizes tubulin, to an extent much greater than does colchicine alone. IKP104 appears to have two classes of binding site on tubulin, differing in affinity; the acceleration of decay appears to be mediated by the low-affinity site (Chaudhuri et al., 1998, J. Protein Chem., in press). We investigated the relationship of the binding of IKP104 and vinblastine. We found that the high affinity site or sites of IKP104 overlap with or interact with the vinblastine binding sites, but that the low-affinity site is distinctly different. PMID- 9853685 TI - Importance of long-range interactions in (alpha/beta)8 barrel fold. AB - Protein structures are stabilized by both local and long-range interactions. In this work, we analyzed the importance of long-range interactions in (alpha/beta)8 barrel proteins in terms of residue distances. We found that the residues occurring in the range of 21-30 residues apart contribute more toward long-range contacts. Indeed, about 50% of successive strands in these proteins are found to occur at a sequential distance of 21-30 residues. The aromatic amino acid residues Phe, Trp, and Tyr prefer the 4-10 range and all other residues prefer the 21-30 range. Hydrophobic-hydrophobic residue pairs are the most preferred ones for long-range interactions and they may play a key role in the folding and stabilization of (alpha/beta)8 barrel proteins. PMID- 9853686 TI - The constituent tryptophans and bisANS as fluorescent probes of the active site and of a secondary binding site of stromelysin-1 (MMP-3). AB - The active site of the catalytic domain of stromelysin-1 (matrix metalloproteinase-3, MMP-3) was probed by fluorescence quenching, lifetime, and polarization of its three intrinsic tryptophans and by the environmentally sensitive fluorescent reporter molecule bisANS. Wavelength-dependent acrylamide quenching identified three distinct emitting tryptophan species, only one of which changes its emission and fluorescence lifetime upon binding of the competitive inhibitor Batimastat. Significant changes in the tryptophan fluorescence polarization occur upon binding by any of the three hydroxamate inhibitors Batimastat, CAS108383-58-0, and Celltech CT1418, all of which bind in the P2'-P3' region of the active site. In contrast, the inhibitor CGS27023A, which is thought to bind in the P1-P1' region, does not induce any change in tryptophan fluorescence polarization. The use of the fluorescent probe bisANS revealed the existence of an auxiliary binding site extrinsic to the catalytic cleft. BisANS acts as a competitive inhibitor of stromelysin with a dissociation constant of Ki=22 microM. In addition to this binding to the active site, it also binds to the auxiliary site with a dissociation constant of 3.40+/-0.17 microM. The auxiliary site is open, hydrophobic, and near the fluorescing tryptophans. The binding of bisANS to the auxiliary site is greatly enhanced by Batimastat, but not by the other competitive inhibitors tested. PMID- 9853687 TI - Amino acid sequence of piratoxin-I, a myotoxin from Bothrops pirajai snake venom, and its biological activity after alkylation with p-bromophenacyl bromide. AB - The complete sequence of the 121 amino acid residues of piratoxin-I (PrTX-I), a phospholipase A2 (PLA2)-like myotoxin from Bothrops pirajai snake (Bahia jararacussu) venom, is reported. From the sequence, an M, of 13,825 and an approximate pI of 8.3 were calculated. PrTX-I shows a high sequence homology with Lys-49 myotoxins from other bothropic (approximately 95%) and nonbothropic (approximately 80%) venoms, but only 70-75% homology when aligned with the catalytically active Asp-49 PLA2s. When compared with bothropstoxin-I from Bothropsjararacussu, which is morphologically almost identical to B. pirajai, only two changes out of 121 total amino acid residues have been observed. The approximate minimal lethal dose LD50 (mice, i.p., 24 hr) of PrTX-I was 8 (6.8 9.1) mg/kg, and the minimal edematogenic dose (MED) in a rat paw model was 39.5+/ 1.8 ug. After alkylation of His-48 with p-bromophenacyl bromide, the MED was 40.1+/-1.9 ug, but up to 4 LD50 were unable to cause death in any of a group of eight mice after 72 hr. Therefore the edematogenic activity was retained and apparently did not involve His-48, suggesting that at least two biologically active sites are present in PrTX-I. PMID- 9853688 TI - Characterization of a subunit structure and stability of the recombinant porin from Neisseria gonorrhoeae. AB - An outer membrane PIA protein from Neisseria gonorrhoeae strain FA19 was expressed in Escherichia coli and refolded in vitro in the presence of zwitterionic detergent. Its proper folding and subunit organization was confirmed by comparison with the native counterpart. The unfolding of PIA has been investigated using fluorescence spectroscopy and analytical size-exclusion chromatography methods. Analysis of the denaturation pathway of the PIA revealed that it forms an unusually labile quaternary structure. In the presence of 1 M guanidinium chloride (GdmCl) or upon heating up to 50 degrees C, dissociation of the PIA oligomer was observed resulting in the formation of folded monomeric intermediates. Unfolding of monomers occurs at 80 degrees C or in the presence of 4.3 M GdmCl, indicating high intrinsic stability toward both GdmCl and elevated temperatures. Both oligomeric and monomeric forms of PIA exhibited affinity to the hydrophobic probe 1-anilinonaphthalene-8-sulfonic acid (ANS) and bind with Kd=80 and 130 microM, respectively. Denaturation of the PIA completely abolished affinity to ANS, suggesting that hydrophobicity is a property of the folded state of the porin. PMID- 9853689 TI - Weight gain in adolescents treated with risperidone and conventional antipsychotics over six months. AB - Weight gain is a serious side effect of antipsychotic therapy. Potential consequences of obesity include noncompliance with medications as well as significant morbidity and mortality. Because atypical antipsychotic agents are increasingly being used in adolescents, it is important to determine and further characterize the rate of weight gain in this population. This study compares weight gain associated with the use of conventional antipsychotic agents with that seen with risperidone in adolescents. This retrospective study identified 60 adolescent inpatients who were taking risperidone (n = 18), conventional neuroleptics (n = 23), or no antipsychotic medication (n = 19). Monthly height and weight measurements were recorded over a 6-month period, allowing body mass index (BMI) values to be calculated. Subjects treated with risperidone gained a mean of 8.64 kg and increased in body mass by 3.67 kg/m2 over the 6-month observation period. Those taking conventional antipsychotics gained 3.03 kg or 0.31 kg/m2 in body mass, whereas the control group actually lost weight (-1.04 kg, or -1.01 kg/m2) in the same time frame. Although both antipsychotic groups gained in weight and BMI, the risperidone-treated group gained significantly more body mass than did the conventional antipsychotic group (p = 0.0011). Gains in body weight did not correlate with dose, and concomitant medications such as stimulants and lithium did not contribute to changes in BMI. PMID- 9853690 TI - A review of developmental aspects of cytochrome P450. AB - This article surveys the development of human hepatic P450 cytochromes (CYPs) involved in xenobiotic metabolism from the fetus through the life span and explores possible clinical consequences of developmental issues. These hepatic P450 CYPs come "on line" at different times during fetal and infant development, and each one is discussed in that temporal sequence. CYP3A7. the major fetal hepatic cytochrome, is present during organogenesis, and it is involved in steroid metabolism. Variably expressed in some fetuses, CYP3A5 is also present at significant levels in about half of all children. In adults, CYP3A4 is the major functional member of the CYP3A subfamily. CYP1A1 is also present during organogenesis, and it metabolizes exogenous toxins, some of which are procarcinogens. CYP2E1 may be present in some second-trimester fetuses, and it may be involved in prenatal alcohol metabolism. After birth, hepatic CYP2D6 and CYP2C8/9 and CYP2C18/19 become active. Both CYP2D6 and CYP2C19 have genetic polymorphisms that can bring about differing capacities to metabolize exogenous drugs, including psychotropic drugs. CYP1A2 becomes active in the fourth to fifth postfetal months. It provides the best current examples of the importance of developmental changes in xenobiotic-metabolizing P450 CYPs through its metabolism of caffeine and theophylline in premature infants, neonates, and adolescents. PMID- 9853691 TI - Prescription of neuroleptics for children and adolescents in Italy. AB - The objective of this study was to describe the use of neuroleptics among children and adolescents in the province of Rome. Subjects 5 to 19 years of age who received neuroleptic prescriptions within the National Health Service between 1986 and 1991 were identified. Prevalence and incidence of use were estimated. The yearly prevalence of use during the study period ranged from 1.3 to 1.7 per 1000 inhabitants. The incidence during 1989 was 1.1 per 1000, with a marked increase with age. Haloperidol was the drug most frequently prescribed; 59.2% of the subjects received only one prescription during the year. Among subjects with more than one prescription, 42.3% received neuroleptics in combination. The observed pattern of use suggests that in current practice neuroleptics are also used for the treatment of nonpsychotic conditions. The analysis of data derived from a prescription monitoring system may provide an important contribution to the description of neuroleptic use among children and adolescents. PMID- 9853692 TI - Mixed mania associated with tricyclic antidepressant therapy in prepubertal delusional depression: three cases. AB - This report describes the sudden appearance of mixed mania in three children with delusional depression soon after the commencement of tricyclic antidepressant therapy. The observation is consistent with reports linking psychomotor abnormalities and psychosis in severely depressed juveniles to an increased propensity for manic switching, as well as adult studies that report a greater risk of antidepressant-induced cycling in bipolar compared with unipolar affective illness. The cases described suggest the need for caution when considering tricyclic pharmacotherapy in juveniles with severe depressive disease. PMID- 9853693 TI - Sertraline treatment of mood disorder associated with prednisone: a case report. AB - The use of corticosteroids, particularly high-potency steroids, for the treatment of various inflammatory conditions has been frequently associated with mood disturbance and psychosis. We report on a 12-year-old white boy treated with high doses of prednisone chronically for 7 years who presented with severe depression, irritability, violence, and psychosis. Sertraline was used to treat depressive as well as psychotic symptoms without the use of antipsychotics. This successful treatment of steroid-induced mood disorder and psychosis with a serotonin reuptake inhibitor is consistent with the literature describing a decrease in central and peripheral serotonin secretion due to steroids, as well as a possible relationship between mood and psychotic symptoms and low cerebrospinal fluid serotonin levels. PMID- 9853694 TI - An adolescent male with multiple paraphilias successfully treated with fluoxetine. AB - Little is known about the psychiatric diagnosis and treatment of adolescents who sexually offend. We therefore describe an adolescent sex offender who met DSM-IV criteria for multiple paraphilias (except for the age criterion), bipolar type II disorder, and OCD, whose paraphilic urges and behaviors, depression, and violent obsessions responded to open label fluoxetine after failing to respond to long term residential treatment. Although only a single and uncontrolled observation, this case suggests that some adolescent sex offenders may in fact have paraphilias, and that paraphilias in adolescents, like those in adults, may respond to serotonin reuptake inhibitors. PMID- 9853695 TI - Estimating imprecision profiles in biochemical analysis. AB - We describe a computer program IMPROFIL which determines an imprecision profile of an analytical method from replicated measurements of samples. It calculates the variance function, the coefficient of variation, the power of definition, the critical limit, the limit of detection and the lower limit of quantification. The primary property, the variance function, is determined by two alternative methods: the conventional maximum approximate conditional likelihood method and the newly developed weighted absolute deviation method. For all quantities, confidence intervals are obtained using the bootstrap procedure. The program combines the use of robust numerical techniques, user-friendliness and integration into a spreadsheet program for data pre- and post-processing. The algorithms used are described in detail. Tests with synthetic data sets are used to validate the method and to establish its powers and limitations. Finally, its application to a practical analytical task (tumor marker CA 15-3 in human sera) is reported. For the method to yield a reliable estimate of the variance function and the derived properties, certain minimum requirements on the raw data must be met: They have to be spread throughout the concentration range of interest, there should not be less than three replicates per specimen, and there must be at least of the order of 25 (better at 50) specimens. PMID- 9853696 TI - Influence of fasting on circulating levels of alpha-tocopherol and beta-carotene. Effect of short-term supplementation. AB - We investigated the influence of fasting on the levels of alpha-tocopherol in plasma, erythrocytes and platelets, and on plasma beta-carotene. Six apparently healthy adults were subjected to 17-h feed-fasting experiments at various days before, during and after supplementation with alpha-tocopherol (455 mg/day, 41 days) and beta-carotene (25 mg/day, 24 days). Adipose tissue alpha-tocopherol and beta-carotene were measured at regular intervals. Supplementation increased alpha tocopherol and beta-carotene in all compartments, except for beta-carotene in adipose tissue. Discontinuation caused a rapid return to baseline, except for adipose tissue alpha-tocopherol and plasma beta-carotene. Fasting caused linear increases of free fatty acids, consistent (but small) increases of plasma alpha tocopherol and inconsistent increases of plasma beta-carotene. There were no fasting-related changes in other compartments. We conclude that fasting is unable to increase alpha-tocopherol and beta-carotene in circulating lipoproteins and cells to a considerable extent, both at baseline levels and after short-term supplementation. Maintenance of high levels may necessitate regular high oral intakes. PMID- 9853697 TI - Effects of postprandial hyperlipemia on the vitamin E content of lipoproteins. GERBAP section Lipoproteines. Groupe d'Evaluation et de Recherche de l'Assistance Publique des Hopitaux de Paris. AB - Delayed postprandial clearance of triglyceride-rich lipoproteins (TGRL) could induce a decrease of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) vitamin E content through its transfer into TGRL. We thus studied lipoprotein vitamin E content during postprandial hypertriglyceridemia induced by a high fat meal without vitamin E supplement. Venous blood was drawn from five healthy male volunteers following a 12 h fast at (t0) then 2 h (t2), 4 h (t4), 6 h (t6) and 8 h (t8) after a 80 g fat meal. In plasma, only TG significantly varied during the postprandial period with a large interindividual variability. Mean composition of lipoproteins in terms of mass was not significantly modified. The amount of vitamin E significantly increased in TGRL and decreased in LDL plus HDL at t4 and t6 relative to t0. Vitamin E content of TGRL and LDL but not of HDL decreased significantly at t4. The mean decrease was 20% (range 5%-54%) for the LDL. LDL- and HDL vitamin E content correlated inversely with plasma TGRL levels. Our data suggest that LDL from subjects with delayed chylomicron clearance could be less protected against oxidation. PMID- 9853698 TI - Glomerular filtration rate determination in diabetic patients using iohexol clearance--comparison of single and multiple plasma sampling methods. AB - Use of iohexol clearance has been described as the gold standard for the measurement of glomerular filtration rate (GFR). It is suggested that multiple plasma sampling following iohexol injection is required to accurately determine GFR by area under plasma clearance curve. The aim of this study was to determine whether single plasma sampling 4 h after injection of iohexol could accurately determine GFR in diabetic patients with mild to moderate renal failure, compared to multiple plasma sampling. A total of 120 GFR determinations in 36 patients with non-insulin dependent diabetic renal disease were done over 1 year. No acute deterioration was seen in renal function following injection of contrast in any patient. Strong correlation in GFR measurement was observed between the multiple plasma sampling method and the single plasma sampling method (r2 = 0.975). Single plasma sampling 4 h after bolus injection of iohexol is a safe and accurate method of determining GFR and change in GFR in diabetic subjects with mild to moderate renal impairment. PMID- 9853699 TI - Comparison of advanced glycation endproducts on haemoglobin (Hb-AGE) and haemoglobin A1c for the assessment of diabetic control. AB - Glycation process in vivo results in two different products: early and advanced glycation endproducts (AGEs). The mechanism of early product formation has been well described, with HbA1c as the best-studied example. The finding that advanced glycation endproducts are also formed on haemoglobin suggests that HbA1c is a precursor for Hb-AGE formation. HbA1c has been well established as an important indicator for glycaemia monitoring, but the diagnostic role of Hb-AGE has not yet been clarified. A question is whether HbA1c and Hb-AGE are competitive or complementary parameters. In our study, Hb-AGE was quantified by the competitive ELISA technique using polyclonal anti-AGE-RNase antibodies to detect AGE immunoreactivities of proteins precipitated in red cell hemolysate. Results are expressed as AGE units/mg Hb. Hb-AGE was analysed in three groups of patients divided according to HbA1c values as follows: group I (n = 25) HbA1c < 7%, Hb-AGE = 6.93 (5.7-7.3) U/mg; group II (n = 25) HbA1c = 7-10%, Hb-AGE = 8.62 (7.7-10.2) U/mg; and group III (n = 25) HbA1c > 10%, Hb-AGE = 12.47 (10.8-13.9) U/mg (median (interquartile range)). A close relation between the amounts of red cell HbA1c and Hb-AGE was observed in all diabetic subjects (n = 75) r = 0.77, P < 0.001. Patients with HbA1c level > 8% were considered to be in poor glycaemic control and those with HbA1c < 8% in good control. In the well-controlled subgroup (n = 33), HbA1c and Hb-AGE were less tightly correlated (r = 0.37, P <0.001). However, in those patients with a higher level of HbA1c = 12.55 (8.9-13.3)% (n = 42), the related Hb-AGE was 11.5 (10.3-12.8) U/mg Hb, yielding a more significant correlation (r = 0.51, P < 0.001). The content of Hb-AGE did not correlate with age (r = 0.09), diabetes duration (r = 0.05) or severity of retinopathy and/or nephropathy. The observed difference may reflect a different kinetic rate of HbA1c production and subsequently the rate of Hb-AGE formation. The discrepancy in the correlation between HbA1c and Hb-AGE suggests that they are complementary rather than opposed parameters. The amount of haemoglobin-linked AGEs does not correlate with the presence or absence of retinopathy and/or nephropathy. It seems that Hb-AGE represents only the metabolic status, equally in the subjects with and without diabetic microangiopathy. PMID- 9853700 TI - Comparison of two immunoradiometric assays for parathyroid hormone-related protein in the evaluation of cancer patients with and without hypercalcemia. AB - Hypercalcemia is a common paraneoplastic syndrome due to the secretion by tumors of parathyroid hormone-related protein (PTHrP) and/or other osteolytic factors. In the present study, we have measured plasma PTHrP using two immunoradiometric assays for PTHrP, assay N (Nichols) and assay I (INCSTAR), recognizing the 1-86 domain of PTHrP, for the evaluation of malignancy-associated hypercalcemia. The study included 25 tumor patients with hypercalcemia (HCa) [corrected serum calcium (SCa) > or = 2.70 mmol/L], 20 normocalcemic patients with cancer (NCa), and ten healthy control subjects. Plasma PTHrP was either undetectable or within the respective normal range in the majority of NCa patients and in the control subjects, with both assays. Plasma PTHrP was increased in 13 and 15 of HCa cases with assay N and assay I, respectively. PTHrP was elevated in plasma in 5/6 (assay N) and 3/6 (assay I) HCa patients with squamous tumors. However, plasma PTHrP was high in only 2/9 (assay N) and 1/9 (assay I) HCa cases with hematological tumors. Less than 40% of HCa patients with bone metastases, and >75% of those without bone involvement, had elevated plasma PTHrP with both assays. Detectable plasma PTHrP and SCa were significantly correlated using assay N (p = 0.025) and assay I (p = 0.01), in the HCa group. A highly significant correlation (p <0.001) was found between detectable plasma PTHrP with both assays, and a high agreement between them based on simple kappa statistics (p < 0.001), in the latter group. Our results indicate that each assay may be similarly useful in detecting PTHrP hyperproduction in cancer patients. PMID- 9853701 TI - Portal but not peripheral serum levels of interleukin 6 could interfere with glucose metabolism in patients with pancreatic cancer. AB - Interleukin 6 (IL-6), an autocrine growth factor for many tumors, seems to favour tumor spread to the liver. Our aims were first to evaluate the pattern of portal and systemic IL-6 levels in patients with pancreatic cancer (PC, n = 18) and chronic pancreatitis (CP, n = 22) compared with controls (CS, n = 20); and second, to ascertain whether there was any relation between IL-6 levels and tumor spread or PC-associated Diabetes mellitus. For all subjects, a fasting serum sample was obtained from a cubital vein; a portal serum sample was obtained from nine PC and three CP patients. In cubital and portal sera we measured IL-6, interleukin 1 beta (IL-1b), CA 19-9, c-reactive protein (CRP) and amylase. Systemic IL-6 levels were significantly higher in PC patients than in CS. In PC, portal IL-6 levels were significantly higher than the corresponding systemic values. The same pattern was found in the three CP patients, whereas IL-1b, CA 19 9, CRP and amylase portal levels were the same as systemic values. No correlation was found between PC stage and systemic or portal IL-6 levels. Portal IL-6 levels were correlated with the corresponding fasting serum glucose values. A significant correlation was found between IL-6 values and CRP, ALT, total bilirubin, GGT and creatinine, but not amylase. IN CONCLUSION: (1) Portal IL-6, which is partly of pancreatic origin, is first metabolised in the liver; (2) Systemic IL-6 reflects hepatic and renal functions rather than local conditions in the pancreas; (3) IL-6 does not appear to influence PC spread; (4) IL-6, which is released in large amounts by the inflamed pancreas, may contribute to determining diabetes, thus interfering with the signal transducing pathways involved in glucose metabolism in liver cells. PMID- 9853702 TI - Differential expression of double-band apolipoprotein(a) phenotypes in healthy Spanish subjects detected by SDS-agarose immunoblotting. AB - A sodium dodecyl sulphate-agarose apolipoprotein(a) [apo(a)] phenotyping method was set up to attain accurate scanning densitometry of proteins. Serum samples from 99 healthy Spanish men were analysed and twenty-five different apo(a) isoforms (12 to 37 kringle 4 repeats) were detected. Double-band phenotypes accounted for 39.4% (n = 39) and three different patterns of protein expression were identified: pattern A (20.5% of double-band phenotyped samples) predominantly expressed the highest molecular weight isoform; pattern B (53.9%) mainly the lowest molecular weight isoform, and pattern AB (25.6%), expressed both isoforms equally. A significant linear association between expression pattern and lipoprotein(a) [Lp(a)] concentration > or = 0.30 g/l was observed. Single-band phenotyped samples (n = 60) were stratified according to apo(a) kringle 4 repeat categories and showed that 90% of isoforms < 20 K4 repeats had high Lp(a) concentrations (> or = 0.30 g/l), whereas isoforms with 20 to 24 or more than 24 kringle 4 repeats had Lp(a) concentrations > or = 0.30 g/l in 47% and 14%, respectively. A logistic regression model was fitted to test the association between apo(a) size, expression pattern and Lp(a) concentration. In this model, apo(a) isoform < 25 kringle 4 repeats was significantly associated with serum Lp(a) concentration > or = 0.30 g/l in both single and double-band phenotyped samples (odds ratio = 8.9, p < 0.001). In the latter, a differential expression pattern with respect to smaller size isoforms (pattern AB vs A) was significantly associated with Lp(a) concentration > or = 0.30 g/l (odds ratio = 17.97, P = 0.045). Heterogeneity in protein apo(a) size expressed according to kringle 4 repeat number could be categorized in heterozygous phenotypes as three patterns. When small-sized isoform was expressed (pattern B) or both isoforms were equally expressed (pattern AB), the probability of having Lp(a) > or = 0.30 g/l is higher. PMID- 9853703 TI - Cocaine induced T cell proliferation in the rat: role of amygdala dopamine D1 receptors. AB - The immunomodulatory effects of local administration of cocaine into the amygdala were studied in the rat. Intra-amygdala infusion of cocaine significantly and dose-dependently increased the proliferative response of splenocytes to concanavalin A (Con A). A similar effect on the immune response was also observed in rats, microinfused into the central amygdala with the selective D1 receptor agonist SKF 38393. The increase of the proliferative response of splenocytes to Con A was inhibited by coinfusion within the central amygdala of the dopamine D1 receptor antagonist SCH 23390, together with cocaine, but not by coinfusion of the dopamine D2 receptor antagonist eticlopride. These results suggest that cocaine may produce at least some of its effects on the immune system through the activation of brain dopamine neurotransmission and that the central amygdala may represent a critical structure mediating cocaine-induced T cell proliferation. PMID- 9853704 TI - Simultaneous assay of ornithine decarboxylase and polyamines after central nervous system injury in gerbil and rat. AB - Ornithine decarboxylase (ODC) is considered the rate-limiting enzyme in polyamine biosynthesis. An increase in putrescine (a natural polyamine) synthesis after central nervous system (CNS) injury appears to be involved in blood-brain barrier dysfunction, development of vasogenic edema and neuronal death. An improved method is described to determine the ODC activity as well as polyamine levels from the same brain tissue. The polyamine results showed no significant differences from data obtained with the conventional assay. The advantages of this method are to: (1) minimize the number of animals needed for the study, and (2) eliminate any internal inconsistencies resulting from use of two independent groups of animals for ODC and polyamine measurements. Using this method, ODC activities and polyamine levels were measured in cortices and hippocampi from global transient ischemia of gerbils and traumatic brain injury (TBI) of rats. PMID- 9853705 TI - Differences between 'congenital mirror movements' and 'associated movements' in normal children: a neurophysiological case study. AB - In this study we analysed how far physiological associated movements in normal children (which may be present up to the age of 10 years) share the same physiological mechanism with clinically apparent mirror movements. Transcranial magnetic stimulation (TMS) and kinematic movement analysis were applied in a 4 year-old child with congenital mirror movements (CMM). The results were compared with a normative data base of clinically normal children. In the child with CMM focal TMS of one motor cortex induced bilaterally symmetrical responses in distal and proximal upper extremities muscles with identical ipsi- and contralateral latencies. Also kinematic analysis showed a precise symmetrical onset of intended and unintended contralateral movements, whereas normal children with associated movements showed a variable movement onset delay between extremities. The data suggest a different physiological mechanism underlying these two varieties of elementary associated motor activity in childhood. PMID- 9853706 TI - Isoform-specific vasoconstriction induced by apolipoprotein E and modulation of this effect by Alzheimer's beta-amyloid peptide. AB - Abeta peptides are thought to be centrally involved in Alzheimer's disease (AD) pathogenesis, although Abeta's pathophysiological mechanisms remain to be elucidated. We previously showed that soluble beta-amyloid1-40 (Abeta) and Abeta1 42 exhibit vasoactive properties, and are able to promote vasoconstriction in rat aortae induced by an endogenous vasoconstrictor, endothelin-1. It is well established that the APOE epsilon4 allele confers risk for both familial and sporadic AD, as well as for hypertension. We now report that physiologic amounts (10 nM) of specific human recombinant apoE isoforms are vasoactive (E4 > E3, and not E2) in isolated rat aortae. In order to investigate if various apoE isoforms could modulate Abeta vasoactivity, we co-incubated Abeta1-40 with various isoforms of apoE in our tissue bath system. Our results show that, while none of the APOE isoforms are able to affect the maximum constriction induced by Abeta; the apoE E4 isoform synergistically enhances the rate of vasoconstriction induced by Abeta. Our data suggest that apoE may promote hypertension and contribute to AD pathogenesis via enhancement of vasoconstriction, and support a link between hypertension, cerebral amyloid angiopathy and AD. PMID- 9853707 TI - Evoked quantal currents at neuromuscular junctions of wild type Drosophila larvae. AB - Evoked excitatory postsynaptic currents (EPSC) were recorded with an extracellular macropatch electrode from glutamatergic neuromuscular junctions of Drosophila larvae. At 20 degrees C quantal current amplitude was about -400 pA and the 10-90% rise time was slightly below 0.2 ms for the fastest rising events and on average 0.3+/-0.1 ms in our best recordings. The quantal currents often had 'shoulders' but decayed approximately monoexponentially from half amplitude. The time constant of the exponential fit varied with mean values ranging from 2.5 ms to 7.7 ms in 13 experiments and an average value of 4.4+/-1.6 ms. Comparison of these results with data obtained earlier with outside-out patches of larval muscle membrane (Heckmann, M. and Dudel, J., Biophys. J., 72 (1997) 2160-2169) leads us to conclude that glutamate has to reach a saturating peak concentration of at least 10 mM in the synaptic cleft to allow the observed short quantal current rise times. To account for the time course of the quantal current decay one has to assume that the glutamate concentration in the synaptic cleft remains in the millimolar range for more than a millisecond and that the time course of the decay of the quantal currents is in part due to desensitization of the postsynaptic receptor channels. PMID- 9853708 TI - A comparative study of the beta-adrenoceptors in higher visual centres of birds. AB - This report describes the distribution of beta-adrenergic receptors in the telencephalic visual nuclei of chick, duck, pigeon, parakeet and goldfinch using [3H]CGP 12177 (4-3-t-butylamino-2-hydroxypropoxy-[5,7(3)H] benzimidazol-2-one) as a radioligand. The results reveal a predominance of the beta2-adrenoceptor subtype in all the species studied and that this subtype fits the pharmacological profile described for mammals. It is also demonstrated that the autoradiographic interspecific differences described in previous studies are due to changes in Bmax, while KD values remain in a similar range (0.1-0.6 nM). The distribution of beta-adrenoceptors was fairly similar in the areas of the visual Wulst of all five species studied while striking differences were found in the ectostriatum, the higher centre of the tectofugal pathway. Our findings support a role for ectostriatal beta-adrenoceptors in the visual adaptation and evolution of birds. PMID- 9853709 TI - Brain potentials in patients with music perception deficits: evidence for an early locus. AB - Twelve patients with an acute cerebrovascular accident were assigned to a group with music perception deficits (amusia, n = 6) or a group without such deficits (n = 6) on the basis of a new test-battery for music-perception skills. Event related brain potentials (ERPs) were recorded in an auditory classification task designed to elicit several components; the N1 as a correlate of initial auditory cortical processing, the P3a as an index of automatic attentional orienting, and the P3b as a measure for controlled stimulus evaluation. Patients with amusia showed a significant amplitude decrement for the P3a relative to controls and patients without amusia suggesting an impairment of early stimulus evaluation. P3b was reduced in both patient groups relative to control. These data show that amusia is quite common in unselected stroke patients and suggest deficits of generic rather than music-specific cognitive processes as the underlying cause. PMID- 9853710 TI - Nitric oxide synthase containing neurons in the ventral lateral geniculate of the rat project to the optic pretectal nuclei. AB - We combined fluorescent tracing with immunohistochemistry to examine nitric oxide synthase (NOS) containing neurons in the rat ventral lateral geniculate nucleus (vLGN). NOS immunopositive neurons in vLGN had a similar appearance to previously described NADPH-d positive neurons. The majority (96%) of the NOS immunopositive neurons in vLGN projected to the pretectal nuclei and these represented 16% of all the vLGN neurons that project to the pretectal nuclei. No NOS immunopositive neurons projected to the superior colliculus. Co-localization of NADPH-d and gamma-aminobutyric acid (GABA) has been reported in vLGN neurons but we found few cells containing both NOS and GABA. PMID- 9853711 TI - NMDA induced glutamate release from the suprachiasmatic nucleus: an in vitro study in the rat. AB - The suprachiasmatic nucleus (SCN) has been identified as a pacemaker for mammalian circadian rhythms. Excitatory amino acid receptors, especially N-methyl D-aspartate (NMDA) receptors, have been considered to play an important role in the transmission of light information from the retina to the circadian clocks in the SCN. In the present study, we showed that application of NMDA at circadian time (CT) 12-15 induced significant glutamate release from the SCN region in vitro. The NMDA-induced glutamate release was blocked by co-application of the NMDA receptor antagonist MK-801, but not by that of tetrodotoxin. These results suggested that glutamate stimulated its own release by activating NMDA receptors. This NMDA-induced glutamate release through NMDA receptor-mediated mechanisms might be involved in NMDA-induced potent phase shifts. PMID- 9853712 TI - Distribution of acetylcholinesterase in the central nervous system of harvestmen (Arachnida: Opilionida). AB - Enzyme histochemical technique (Gomori and Koelle methods, reaction product development according to Lewis) was applied on 10 microm frozen sections of formalin-fixed material to demonstrate the distribution of acetylcholinesterase in the central nervous system (CNS) of harvestmen (Opilionida: Phalangiidae). Distinctly positive reactions were confined to the neuropil, showing strong staining especially in protocerebral brain centres (optic lobes, cerebral ganglia), the cheliceral ganglia (stomodeal bridge, afferent tracts), and the connective ring systems of the subesophageal nerve mass. The results obtained are discussed with regard to central coordinative functions of the respective brain parts. PMID- 9853713 TI - Differential recruitment of high frequency wavelets (600 Hz) and primary cortical response (N20) in human median nerve somatosensory evoked potentials. AB - Human median nerve somatosensory evoked potentials contain a burst of high frequency (600 Hz) wavelets superimposed on the primary cortical response (N20). These presumably reflect highly-synchronized repetitive thalamic and/or intracortical population spike bursts and are diminished in non-REM sleep with N20 persisting. Here the burst/N20 relation in awake subjects was examined by using eight different intensities of electric median nerve stimuli. In all subjects the amplitude recruitment of both N20 and burst could be modeled adequately as a sigmoidal function of stimulus intensity. While 8/10 subjects showed a parallel recruitment, 2/10 subjects required significantly higher stimulation intensities for burst than for N20 recruitment. This dampened burst recruitment possibly reflects slight vigilance fluctuations in open-eyed awake subjects; a further increase of burst thresholds could explain the burst attenuation when entering shallow sleep. PMID- 9853714 TI - Localization of ATP-gated P2X receptor immunoreactivity in rat sensory and sympathetic ganglia. AB - The distribution of P2X1, P2X2, P2X3, P2X4, P2X5 and P2X6 receptors, a family of ATP-gated cation channels, in rat trigeminal, dorsal root, nodose, superior cervical, and coeliac ganglia was studied immunohistochemically. It was found that polyclonal antibodies for the six P2X receptor subtypes could label over 90% of neurones in these ganglia to different intensities. There was also considerable variation in intensity of immunoreactivity in individual neurones within each ganglion. In dorsal root and trigeminal ganglia expression of the P2X3 receptor was much higher than for the other five subtypes. P2X3 immunoreactivity was much more intense in the small-diameter neurones than in the large-diameter neurones. In nodose ganglion, both P2X2 and P2X3 antibodies labelled the neurones intensely. In sympathetic superior cervical and coeliac ganglia, immunoreactivity to five P2X receptor subtypes were detected (exception, P2X5), with P2X2, P2X4 and P2X6 showing higher intensity. Low level expression of P2X3 receptor in sympathetic ganglia indicates that this receptor subtype is not limited to the sensory ganglia where it was highly expressed. The results have demonstrated that both sensory and sympathetic ganglia express a variety of P2X receptor subtypes and that different subtypes are expressed to different levels and by different subpopulations of neurones. PMID- 9853715 TI - Proliferation-dependent vulnerability of glucose-deprived astrocytes to nitric oxide-induced cytotoxicity. AB - The present study investigated the vulnerability of glucose-deprived astrocytes to nitric oxide (NO)-induced cytotoxicity. Primary murine cortical astrocyte cultures of different ages were deprived of glucose in the presence of the NO releasing reagent 3-morpholinosydnonimine. Glucose-deprived astrocytes displayed an age-dependent vulnerability to NO cytotoxicity. However, this difference in vulnerability with age was found to be dependent on initial plating density. The augmented deaths of astrocyte cultures were consistent with the [3H]thymidine incorporation pattern. Thus, rapidly dividing astrocytes were much more susceptible to NO cytotoxicity under glucose-deprived conditions. The data indicate that the vulnerability of glucose-deprived astrocytes to NO cytotoxicity depends on the proliferative state rather than age in vitro. PMID- 9853716 TI - C2-ceramide attenuates prostaglandin F2alpha-induced vasoconstriction and elevation of [Ca2+]i in canine cerebral vascular smooth muscle. AB - Sphingolipids have emerged as important components of signal transduction pathways involved in a variety of cellular processes. In the present study, we examined the effects of C2-ceramide, a cell-permeable sphingolipid, on contraction of canine cerebral vascular smooth muscle and intracellular free Ca2+ ([Ca2+]i). C2-ceramide (10(-8)-10(-4) M) alone did not elicit any significant changes in either basal tension or resting levels of [Ca2+]i in canine cerebrovascular muscle. However, C2-ceramide (10(-7)-10(-4) M) attenuated prostaglandin F2alpha (PGF2alpha)-induced contractions in isolated canine cerebrovascular smooth muscle rings. C2-ceramide (10(-5) M) inhibited the secondary phasic rise of [Ca2+]i evoked by PGF2alpha in cultured canine cerebral vascular smooth muscle cells, resulting in decreases in the elevation in [Ca2+]i. NO inhibitors (L-NNA, L-NMMA), an inhibitor of prostanoid synthesis (indomethacin), an inhibitor of opiate actions and several inhibitors of the pharmacologic actions of various vasoactive amines all failed to interfere with the vasorelaxant response of C2-ceramide. Our results suggest that the sphingomyelin signaling pathway may play an important regulatory role in cerebral arterial wall tone. PMID- 9853717 TI - Humoral signals mediate the circadian expression of rat period homologue (rPer2) mRNA in peripheral tissues. AB - Northern blot analysis revealed the circadian expression of rat period homologue (rPer2) mRNA in peripheral mononuclear leukocytes that have no neuronal connections. The mRNA expression in the suprachiasmatic nucleus (SCN), peripheral mononuclear leukocytes, and other peripheral tissues exhibited prominent, synchronous circadian oscillation with peaks at early night. As we have previously shown the peripheral circadian expression of rPer2 mRNA is abolished in SCN-lesioned rats [Sakamoto, K., Nagase, T. Fukui, H., Horikawa, K., Okada, T., Tanaka, H., Sato, K., Miyake, Y., Ohara, O., Kako, K. and Ishida, N., J. Biol. Chem., 273 (1998) 27039-27042], our findings suggest that some humoral signals, driven by the SCN, mediate the circadian expression of mammalian per homologues in peripheral tissues. PMID- 9853718 TI - Acute morphine dependence in mice selectively-bred for high and low analgesia. AB - Acute morphine dependence was compared in mice selectively-bred for high (HA) and low (LA) swim stress-induced analgesia and high (HAR) and low (LAR) levorphanol analgesia by counting the number of naloxone-precipitated jumps. Whereas LAR mice displayed greater acute morphine dependence than HAR mice, HA and LA mice did not differ. No genotypic differences were observed in non-dependent mice, discounting possible differences in basal naloxone sensitivity and/or opioid peptide levels. Thus, the two selection projects, while both producing lines exhibiting highly divergent sensitivity to morphine analgesia, have not had analogous effects on all opioid measures, supporting the notion of independent genetic mediation of opioid analgesia and dependence. Further, these data suggest that analgesic sensitivity may not predict sensitivity to morphine dependence. PMID- 9853719 TI - Diagnosis and treatment of common metabolic spinal disorders in the geriatric population. AB - BACKGROUND: Bone is constantly resorbed and remodeled throughout life. After approximately age 30, there is a net loss of bone mass. This places the geriatric population at an increased risk of pathologic bone disorders that can lead to fractures and deformity. METHODS: In this paper, we review bone metabolism and remodeling and introduce the proper diagnostic techniques. The most common pathologic spinal disorders are introduced, with emphasis on presentation and treatment options. RESULTS: To prevent excessive bone loss, patients should be educated on proper nutrition (calcium and vitamin D requirements) and lifestyle (avoiding alcohol and cigarette smoking). Sex hormone and drug therapies are available to reduce bone loss. New bisphosphonates such as alendronate sodium (Fosamax) have been effective in increasing bone mass. CONCLUSIONS: Early diagnosis and proper treatment of pathologic bone disorders can reduce the incidence of fracture and allow the patient a more productive and comfortable life. PMID- 9853720 TI - Pulmonary contusion in children: diagnosis and management. AB - BACKGROUND: Pulmonary contusion (PC) is the most common injury identified in pediatric blunt chest trauma. The disagreement over management principles of children with PC may complicate care of these patients. METHODS: We reviewed the literature and our institutional experience. RESULTS: Pulmonary contusion is most often diagnosed by chest roentgenogram, with computed tomography of the chest reserved for more precise evaluation of the mediastinum and aorta. Hypoxemia and respiratory mechanical insufficiency are the most common causes of morbidity from early chest injury. Complications include pneumonia and the adult respiratory distress syndrome, which together may occur in up to one half of all cases. CONCLUSIONS: Early diagnosis, maintaining euvolemia, and selective mechanical ventilation may help to limit morbidity. Management guidelines based on the available literature may improve the care of these patients. PMID- 9853721 TI - Hidradenitis suppurativa. AB - BACKGROUND: Hidradenitis suppurativa (HS) is a recurrent, suppurative disease manifested by abscesses, fistulas, and scarring. METHODS: We reviewed the literature to identify reliable information regarding epidemiology, pathogenesis, clinical manifestations, evaluation and differential diagnosis, treatment, complications, and prognosis. RESULTS: Hidradenitis suppurativa usually affects young women, with a prevalence of 0.3% to 4% in industrialized countries. Once considered to be "apocrine acne," HS is actually a defect of terminal follicular epithelium. Obesity, chemical irritants, or hyperandrogenism are not consistently associated; bacterial involvement is secondary. Hidradenitis suppurativa should be suspected in young adults with recurrent, deep furuncular lesions in flexural sites, especially when such lesions respond poorly to antibiotic therapy. Clindamycin and isotretinoin may be useful, though wide excision with healing by granulation is considered most efficacious. Anemia, arthropathy, and squamous cell carcinoma are potential complications. CONCLUSIONS: Since spontaneous resolution is rare and progressive disability the rule, early definitive surgical treatment of HS is advisable. PMID- 9853722 TI - Traditional and evidence-based acupuncture: history, mechanisms, and present status. AB - BACKGROUND: In 1983, the Southern Medical Journal advised its readers that a scientific basis might underlie the popular practice of ancient Chinese acupuncture. Recent studies have proven this to be correct, and a 1997 National Institutes of Health consensus panel recommended acupuncture as a useful clinical procedure. METHODS: Pertinent articles in the literature were reviewed, including our own research. Significantly, we had access to recent important studies from China. RESULTS AND CONCLUSIONS: Most of the 10,000 acupuncturists in the country today practice metaphysically explained "meridian theory" acupuncture using needles to supposedly remove blockages of a hypothesized substance "Qi." Scientific research has shown that healing is not by manipulating Qi but rather by neuroelectric stimulation for the gene expression of neuropeptides. Needles are not necessary. Evidence-based neuroelectric acupuncture requires no metaphysical rituals. It is a simple, useful clinical tool for pain modulation and other conditions and can be easily taught to physicians. PMID- 9853723 TI - Acupuncture: a review of its history, theories, and indications. AB - BACKGROUND: The National Institutes of Health recently recommended acupuncture as an effective tool for the treatment of various health problems. Acupuncture is an old technique but has been popular in the United States only since 1972. Its history, theories, and indications are not well known to the medical community. METHODS: We reviewed the literature to gather information on the history, techniques, physiology, indications, adverse effects, and opposing views to acupuncture. RESULTS: The mechanism by which acupuncture works involves neurotransmitters and adrenocorticotropic hormones. It appears to be effective in the treatment of pain, nausea, and drug detoxification and in stroke victims. Studies suggest acupuncture is no more effective than placebo. Acupuncture side effects have rarely been reported. CONCLUSIONS: Acupuncture appears to be a safe and effective alternative medical therapy for certain health problems. More controlled research is necessary to better understand the range of its clinical application. PMID- 9853724 TI - Lifetime use of alternative therapy: a study of Florida residents. AB - BACKGROUND: Clinically relevant questions remain about who uses alternative medicine, which treatments they use and why. METHODS: The random digit dialing survey method was used to ask Florida residents about their lifetime use of 11 different alternative therapies. The response rate was 54% (n=1,012). RESULTS: Sixty-two percent of respondents had used one or more of these alternative therapies. Women, unmarried persons, those with regular physicians, and those with poor self-rated health were the highest users. Home remedies were used by 31% of the respondents, followed by special diets (24%), relaxation techniques (20%), and herbal medicines (18%). Acupuncture, biofeedback, energy healing, and hypnosis were used by less than 5% of the sample. CONCLUSIONS: Most respondents used an alternative therapy at some time. Ethnic diversity may be a useful marker for more diversity in alternative therapy. These results affirm the need for accelerated physician education in alternative medicine to help physicians respond appropriately to patients' inquiries about alternative therapy. PMID- 9853725 TI - Low postoperative infection rates with instrumented lumbar fusion. AB - BACKGROUND: Studies have shown postoperative infection rates increase with the use of internal instrumentation. It is believed that longer operative times, prolonged retraction, instrumentation, and bone grafting lead to a higher risk of infection. METHODS: We retrospectively reviewed 126 consecutive instrumented lumbar fusions. All had bone graft. The infection rate was statistically compared with previously reported values. RESULTS: Our infection rate (0.8%) was lower than the combined data from the literature (2.8%), though not significantly different. No significant differences were noted regarding patient or surgical factors. CONCLUSIONS: This review showed that low rates of postoperative infection can be achieved despite patient or surgical factors. We believe that conforming to strict techniques, including copious irrigation and debridement, and having experienced operating room personnel and short operating times will reduce the incidence of postoperative infections. PMID- 9853726 TI - Predictive value of fetal ultrasonography in the diagnosis of a lethal skeletal dysplasia. AB - BACKGROUND: Certain ultrasonographic findings identified in a fetus suspected of having a skeletal dysplasia may be predictive of a lethal outcome. METHODS: We evaluated 27 fetuses suspected of having a skeletal dysplasia using targeted ultrasonography between 16 and 31 weeks' gestation. Clinical examination and skeletal radiography were done after delivery. RESULTS: A skeletal dysplasia was confirmed and a diagnosis established in all but one case. The skeletal dysplasia was lethal in 23 cases and, in each case, the outcome was accurately predicted prenatally; however, three of the infants survived several months. In 11 of the 23 cases (48%), the specific diagnosis was correctly determined before birth. Ultrasonographic findings not considered to reflect a lethal outcome, were accurately predicted in two other cases. In an additional two, sonographic examination suggested a lethal osteochondrodysplasia, though both survived. Findings consistent with a lethal skeletal dysplasia included a femur length < 1st centile, combined with either a bell-shaped thorax, decreased bone echogenicity, or both. Using these criteria provided a positive-predictive value for neonatal deaths of 80% (20/25), and 92% (23/25) if the three that died in infancy were included. CONCLUSIONS: In the fetus suspected of having a skeletal dysplasia, certain findings on targeted ultrasonography frequently are predictive of a lethal outcome; the ability to predict this appears greatest when more than one of these abnormalities is present. PMID- 9853727 TI - Disorders associated with clinically severe obesity: significant improvement after surgical weight reduction. AB - BACKGROUND: Surgical treatment for patients with clinically severe obesity mainly aims to reduce morbidity. METHODS: Sixty-two patients were examined for disorders associated with morbid obesity before and after surgical weight reduction by vertical banded gastroplasty. All patients were followed-up for 12 to 48 months. RESULTS: At the end of the first postoperative year, 84% of the patients had lost at least 50% of their excess weight. Of the 218 weight-related pathologic conditions existing before the operation, 131 (60%) were completely cured, 50 (23%) showed significant improvement, and only 37 (17%) remained unchanged. The same percentages were obtained for patients followed tip for 24, 36, and 48 postoperative months. CONCLUSIONS: Surgical treatment of clinically severe obesity has a significant effect on the health of the patients by eliminating the associated disorders. PMID- 9853728 TI - Pulmonary embolism and malpractice claims. AB - BACKGROUND: Deep venous thrombosis (DVT) with pulmonary embolization (PE) often occurs as an unexpected event with fatal consequences. This provides a setting for malpractice claims. METHODS: We reviewed 160 consecutive malpractice claims submitted by attorneys for medical expert review during the 11-year period ending in 1997. Seven cases involved DVT with PE. RESULTS: Alleged failure to anticipate and reduce the chance of PE was the basis for six of the claims. All six patients were at risk for lower extremity DVT, and one had a history of DVT 6 months earlier. The PE was manifested by sudden death in three cases. The seventh case represented a complication of heparin therapy for PE. CONCLUSIONS: We conclude that risk management for PE should focus primarily on DVT. Physicians should perform and document an examination for DVT whenever there is a history of lower extremity stasis or it is likely to occur. They should also consider documenting a concurring second opinion when making anticoagulant-related decisions. PMID- 9853729 TI - Over-the-counter sympathomimetics: a risk factor for cardiac arrhythmias in pregnancy. AB - More than 170 over-the-counter (OTC) preparations contain a sympathomimetic agent as the active ingredient. Nonprescription medicines are consumed commonly in our society. Phenylpropanolamine, an alpha-adrenergic agonist as well as a popular decongestant, is consumed at a massive volume of 5 billion doses annually. Over the-counter sympathomimetics have been reported to cause hypertension and arrhythmias. Despite the ability to cause these potentially serious adverse effects and the high-volume consumption of these agents, the medical literature until recently has been scant in reporting adverse events. We report symptomatic ventricular arrhythmia and presyncope in a 36-year-old pregnant woman who consumed relatively high doses of two OTC cold remedies simultaneously. Increased physician awareness of the potential side effects associated with OTC sympathomimetics as well as improved level of patient education are needed. Finally, we support the calls for more prominent warning labels on some selected OTC preparations, including OTC sympathomimetics. PMID- 9853730 TI - Disseminated histoplasmosis and Wegener's granulomatosis. AB - Disseminated histoplasmosis is an unusual complication in endemic areas and has been reported in patients with an immunodeficient state, either from human immunodeficiency virus (HIV) infection or transplantation. In the non-HIV population, rare cases of disseminated histoplasmosis have been reported in patients with leukemia and Hodgkin's disease and patients receiving steroid therapy for various diseases. We report a case of disseminated histoplasmosis in a patient with Wegener's granulomatosis. PMID- 9853731 TI - Solitary choroid plexus metastasis from a renal cell carcinoma. AB - Metastatic tumors to the choroid plexus are rare. We report a case of renal cell carcinoma metastasizing to the choroid plexus of the lateral ventricular atrium. This tumor was shown by magnetic resonance imaging, followed by craniotomy and histologic confirmation. Computed tomography of the abdomen showed the primary tumor. We also reviewed 40 years of the literature and found 14 cases of metastasis to the choroid plexus. We discuss the clinical features of this intraventricular metastasis. PMID- 9853732 TI - Neonatal pharyngeal pseudodiverticulum mimicking esophageal atresia. AB - Pseudodiverticulum of the hypopharynx is an infrequent but potentially serious complication of orogastric or nasogastric tube insertion and endotracheal intubation. We report two cases of injury to the hypopharynx resulting in a pseudodiverticulum of the hypopharynx that was initially diagnosed as esophageal atresia. Both cases were managed successfully by conservative therapy. We also review the literature regarding neonatal pharyngeal pseudodiverticulum. PMID- 9853733 TI - Streptococcus bovis bacteremia: unusual complications. AB - Streptococcus bovis bacteremia is known to be related to neoplastic lesions of the colon. We describe a patient with several complications of S bovis bacteremia and adenocarcinoma of the colon--endocarditis, spondylodiskitis, and splenic abscess. We believe this is the eighth known case of endocarditis and diskitis caused by S bovis and the third case of endocarditis and splenic abscess by S bovis in a patient with adenocarcinoma of the colon. PMID- 9853734 TI - Mediastinal lipomatosis. AB - Mediastinal lipomatosis is a benign condition characterized by a large amount of mature adipose tissue within the mediastinum. It widens the mediastinum and may simulate mass lesions, thus leading to diagnostic errors. We describe a new case of a huge mediastinal lipomatosis and review 36 cases previously reported in the literature. Computed tomography has an important role in the diagnosis of this disease. To avoid invasive and unnecessary procedures, this diagnosis should be considered in any patient with Cushing's syndrome. PMID- 9853735 TI - Primary colon cancer without gross mucosal tumor: unusual presentation of a common malignancy. AB - Colon cancer typically arises in the colonic lumen, allowing for endoscopic detection of cancerous and premalignant lesions. In the case presented, a 73-year old man with iron deficiency anemia had two colonoscopies showing only diverticula and internal hemorrhoids. Three years later, when the patient complained of dull, intermittent lower abdominal pain, a third colonoscopy identified diverticula, three adenomatous polyps, and no other abnormality. Computed tomography (CT) of the abdomen revealed a 5 cm x 6 cm pericolic fluid collection, which was later found to communicate with the sigmoid colon. The surgical specimen from a partial colectomy contained a distal sigmoid perforation with a 2.5 cm moderately differentiated adenocarcinoma in the underlying submucosal tissue. There was no gross intraluminal tumor. Histopathology, including immunohistochemistry, was consistent with colonic adenocarcinoma. Primary colon cancer grossly sparing the mucosa is an unusual presentation for this common malignancy. PMID- 9853736 TI - Maternal mortality associated with adult respiratory distress syndrome. PMID- 9853737 TI - Synthetic braids and limited neck extension. PMID- 9853738 TI - Iceland poised to sell exclusive rights to national health data. PMID- 9853739 TI - It will take more than notebooks to stop fraud. PMID- 9853740 TI - An oblique view of climate. PMID- 9853741 TI - Developmental biology. Attractive genetics. PMID- 9853742 TI - Transcription. A lesson in sharing? PMID- 9853743 TI - Excitatory synapses. Is bigger better? PMID- 9853744 TI - Signal transduction. New exchange, new target. PMID- 9853745 TI - Jurgen Aschoff (1913-98) PMID- 9853746 TI - Julesz's joyfulness. PMID- 9853747 TI - Protease helps yeast find mating partners. PMID- 9853748 TI - Moving ahead through differential visual latency. PMID- 9853749 TI - Enhanced long-term potentiation and impaired learning in mice with mutant postsynaptic density-95 protein. AB - Specific patterns of neuronal firing induce changes in synaptic strength that may contribute to learning and memory. If the postsynaptic NMDA (N-methyl-D aspartate) receptors are blocked, long-term potentiation (LTP) and long-term depression (LTD) of synaptic transmission and the learning of spatial information are prevented. The NMDA receptor can bind a protein known as postsynaptic density 95 (PSD-95), which may regulate the localization of and/or signalling by the receptor. In mutant mice lacking PSD-95, the frequency function of NMDA-dependent LTP and LTD is shifted to produce strikingly enhanced LTP at different frequencies of synaptic stimulation. In keeping with neural-network models that incorporate bidirectional learning rules, this frequency shift is accompanied by severely impaired spatial learning. Synaptic NMDA-receptor currents, subunit expression, localization and synaptic morphology are all unaffected in the mutant mice. PSD-95 thus appears to be important in coupling the NMDA receptor to pathways that control bidirectional synaptic plasticity and learning. PMID- 9853750 TI - Selective amplification by auto- and cross-catalysis in a replicating peptide system. AB - Self-replication has been demonstrated in synthetic chemical systems based on oligonucleotides, peptides and complementary molecules without natural analogues. However, within a living cell virtually no molecule catalyses its own formation, and the search for chemical systems in which both auto- and cross-catalysis can occur has therefore attracted wide interest. One such system, consisting of two self-replicating peptides that catalyse each other's production, has been reported. Here we describe a four-component peptide system that is capable of auto- and cross-catalysis and allows for the selective amplification of one or more of the products by changing the reaction conditions. The ability of this system selectively to amplify one or more molecules in response to changes in environmental conditions such as pH or salt concentration supports the suggestions that self-replicating peptides may have played a role in the origin of life. PMID- 9853751 TI - Low-latitude glaciation and rapid changes in the Earth's obliquity explained by obliquity-oblateness feedback. AB - Palaeomagnetic data suggest that the Earth was glaciated at low latitudes during the Palaeoproterozoic (about 2.4-2.2 Gyr ago) and Neoproterozoic (about 820-550 Myr ago) eras, although some of the Neoproterozoic data are disputed. If the Earth's magnetic field was aligned more or less with its spin axis, as it is today, then either the polar ice caps must have extended well down into the tropics-the 'snowball Earth' hypothesis-or the present zonation of climate with respect to latitude must have been reversed. Williams has suggested that the Earth's obliquity may have been greater than 54 degrees during most of its history, which would have made the Equator the coldest part of the planet. But this would require a mechanism to bring the obliquity down to its present value of 23.5 degrees. Here we propose that obliquity-oblateness feedback could have reduced the Earth's obliquity by tens of degrees in less than 100 Myr if the continents were situated so as to promote the formation of large polar ice sheets. A high obliquity for the early Earth may also provide a natural explanation for the present inclination of the lunar orbit with respect to the ecliptic (5 degrees), which is otherwise difficult to explain. PMID- 9853752 TI - Implications of Deltatheridium specimens for early marsupial history. AB - We describe here two new specimens of the mammal Deltatheridium pretrituberculare from the Late Cretaceous period of Mongolia. These specimens provide information on tooth replacement in basal therian mammals and on lower jaw and basicranial morphology. Deltatheroidans, known previously from isolated teeth, partial rostra and jaws from the late Cretaceous of Asia and possibly North America, have been identified variously as eutherians, as basal metatherians (the stem-based clade formed by marsupials and their extinct relatives), or as an outgroup to both eutherians and metatherians. Resolution of these conflicting hypotheses and understanding of the early evolution of the therian lineage have been hampered by a sparse fossil record for basal therians. The new evidence supports metatherian affinities for deltatheroidans and allows a comprehensive phylogenetic analysis of basal metatherians and marsupials. The presence of specialized marsupial patterns of tooth replacement and cranial vascularization in Deltatheridium and the basal phylogenetic position of this taxon indicate that these features are characteristic of Metatheria as a whole. Other morphological transformations recognized here secure the previously elusive diagnosis of Metatheria. The new specimens of Deltatheridium illustrate the effectiveness of fairly complete fossil specimens in determining the nature of early evolutionary events. PMID- 9853753 TI - A role of Ultrabithorax in morphological differences between Drosophila species. AB - The mechanisms underlying the evolution of morphology are poorly understood. Distantly related taxa sometimes exhibit correlations between morphological differences and patterns of gene expression, but such comparisons cannot establish how mechanisms evolve to generate diverse morphologies. Answers to these questions require resolution of the nature of developmental evolution within and between closely related species. Here I show how the detailed regulation of the Hox gene Ultrabithorax patterns trichomes on the posterior femur of the second leg in Drosophila melanogaster, and that evolution of Ultrabithorax has contributed to divergence of this feature among closely related species. The cis-regulatory regions of Ultrabithorax, and not the protein itself, appear to have evolved. This study provides experimental evidence that cis regulatory evolution is one way in which conserved proteins have promoted morphological diversity. PMID- 9853754 TI - HMG-CoA reductase guides migrating primordial germ cells. AB - The enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase is best known for catalysing a rate-limiting step in cholesterol biosynthesis, but it also participates in the production of a wide variety of other compounds. Some clinical benefits attributed to inhibitors of HMG-CoA reductase are now thought to be independent of any serum cholesterol-lowering effect. Here we describe a new cholesterol-independent role for HMG-CoA reductase, in regulating a developmental process: primordial germ cell migration. We show that in Drosophila this enzyme is highly expressed in the somatic gonad and that it is necessary for primordial germ cells to migrate to this tissue. Misexpression of HMG-CoA reductase is sufficient to attract primordial germ cells to tissues other than the gonadal mesoderm. We conclude that the regulated expression of HMG-CoA reductase has a critical developmental function in providing spatial information to guide migrating primordial germ cells. PMID- 9853755 TI - Gain control of NMDA-receptor currents by intracellular sodium. AB - The influx of Na+ is fundamental to electrical signalling in the nervous system and is essential for such basic signals as action potentials and excitatory postsynaptic potentials. During periods of bursting or high levels of discharge activity, large increases in intracellular Na+ concentration ([Na+]i) are produced in neuronal soma and dendrites. However, the intracellular signalling function of raised postsynaptic [Na+]i is unknown. Here we show that [Na+]i regulates the function of NMDA (N-methyl-D-aspartate) receptors, a principal subtype of glutamate receptor. NMDA-receptor-mediated whole-cell currents and NMDA-receptor single-channel activity were increased by raising [Na+]i and channel activity decreased upon lowering [Na+]i; therefore, the activity of NMDA channels tracks changes in [Na+]i. We found that the sensitivity of the channel to Na+ was set by a Src kinase that is associated with the channel. Raising [Na+]i selectively increased synaptic responses mediated by NMDA receptors, but not by non-NMDA receptors. Thus, the change in postsynaptic [Na+]i that occurs during neuronal activity is a signal for controlling the gain of excitatory synaptic transmission. This mechanism may be important for NMDA-receptor dependent plasticity and toxicity in the central nervous system. PMID- 9853756 TI - Epac is a Rap1 guanine-nucleotide-exchange factor directly activated by cyclic AMP. AB - Rap1 is a small, Ras-like GTPase that was first identified as a protein that could suppress the oncogenic transformation of cells by Ras. Rap1 is activated by several extracellular stimuli and may be involved in cellular processes such as cell proliferation, cell differentiation, T-cell anergy and platelet activation. At least three different second messengers, namely diacylglycerol, calcium and cyclic AMP, are able to activate Rap1 by promoting its release of the guanine nucleotide GDP and its binding to GTP. Here we report that activation of Rap1 by forskolin and cAMP occurs independently of protein kinase A (also known as cAMP activated protein kinase). We have cloned the gene encoding a guanine-nucleotide exchange factor (GEF) which we have named Epac (exchange protein directly activated by cAMP). This protein contains a cAMP-binding site and a domain that is homologous to domains of known GEFs for Ras and Rap1. Epac binds cAMP in vitro and exhibits in vivo and in vitro GEF activity towards Rap1. cAMP strongly induces the GEF activity of Epac towards Rap1 both in vivo and in vitro. We conclude that Epac is a GEF for Rap1 that is regulated directly by cAMP and that Epac is a new target protein for cAMP. PMID- 9853757 TI - Functional interaction between InsP3 receptors and store-operated Htrp3 channels. AB - Calcium ions are released from intracellular stores in response to agonist stimulated production of inositol 1,4,5-trisphosphate (InsP3), a second messenger generated at the cell membrane. Depletion of Ca2+ from internal stores triggers a capacitative influx of extracellular Ca2+ across the plasma membrane. The influx of Ca2+ can be recorded as store-operated channels (SOC) in the plasma membrane or as a current known as the Ca2+-release-activated current (I(crac)). A critical question in cell signalling is how SOC and I(crac) sense and respond to Ca2+ store depletion: in one model, a messenger molecule is generated that activates Ca2+ entry in response to store depletion; in an alternative model, InsP3 receptors in the stores are coupled to SOC and I(crac). The mammalian Htrp3 protein forms a well defined store-operated channel and so provides a suitable system for studying the effect of Ca2+-store depletion on SOC and I(crac). We show here that Htrp3 channels stably expressed in HEK293 cells are in a tight functional interaction with the InsP3 receptors. Htrp3 channels present in the same plasma membrane patch can be activated by Ca2+ mobilization in intact cells and by InsP3 in excised patches. This activation of Htrp3 by InsP3 is lost on extensive washing of excised patches but is restored by addition of native or recombinant InsP3-bound InsP3 receptors. Our results provide evidence for the coupling hypothesis, in which InsP3 receptors activated by InsP3 interact with SOC and regulate I(crac). PMID- 9853758 TI - The receptor Msn5 exports the phosphorylated transcription factor Pho4 out of the nucleus. AB - The movement of many transcription factors, kinases and replication factors between the nucleus and cytoplasm is important in regulating their activity. In some cases, phosphorylation of a protein regulates its entry into the nucleus; in others, it causes the protein to be exported to the cytoplasm. The mechanism by which phosphorylation promotes protein export from the nucleus is poorly understood. Here we investigate how the export of the yeast transcription factor Pho4 is regulated in response to changes in phosphate availability. We show that phosphorylation of Pho4 by a nuclear complex of a cyclin with a cyclin-dependent kinase, Pho80-Pho85, triggers its export from the nucleus. We also find that the shuttling receptor used by Pho4 for nuclear export is the importin-beta-family member Msn5, which is required for nuclear export of Pho4 in vivo and binds only to phosphorylated Pho4 in the presence of the GTP-bound form of yeast Ran in vitro. Our results reveal a simple mechanism by which phosphorylation can control the nuclear export of a protein. PMID- 9853759 TI - Crystal structure of the ligand-binding domain of the receptor tyrosine kinase EphB2. AB - The Eph receptors, which bind a group of cell-membrane-anchored ligands known as ephrins, represent the largest subfamily of receptor tyrosine kinases (RTKs). They are predominantly expressed in the developing and adult nervous system and are important in contact-mediated axon guidance, axon fasciculation and cell migration. Eph receptors are unique among other RTKs in that they fall into two subclasses with distinct ligand specificities, and in that they can themselves function as ligands to activate bidirectional cell-cell signalling. We report here the crystal structure at 2.9 A resolution of the amino-terminal ligand binding domain of the EphB2 receptor (also known as Nuk). The domain folds into a compact jellyroll beta-sandwich composed of 11 antiparallel beta-strands. Using structure-based mutagenesis, we have identified an extended loop that is important for ligand binding and class specificity. This loop, which is conserved within but not between Eph RTK subclasses, packs against the concave beta sandwich surface near positions at which missense mutations cause signalling defects, localizing the ligand-binding region on the surface of the receptor. PMID- 9853760 TI - Fetal growth. PMID- 9853761 TI - The performance of screening tests for ovarian cancer: results of a systematic review. AB - OBJECTIVE: To estimate the performance of currently available tests in detecting ovarian cancer in asymptomatic women. METHODS: Systematic review of prospective screening studies. RESULTS: Twenty-five studies were identified: sixteen studied women at average risk and nine studied women at higher risk. Most studies evaluated only one screening method, were small, detecting few cancers, and gave few follow up details. Sensitivity estimates are therefore imprecise. In a typical larger study, reported sensitivity of ultrasound screening at one year was around 100% (95% CI 54%-100%), while the sensitivity of CA125 measurement followed by ultrasound (multimodal screening) was about 80% (95% CI 49%-95%). False positive rates ranged between 1.2% and 2.5% for grey scale ultrasound, between 0.3% and 0.7% for ultrasound with colour Doppler and between 0.1% and 0.6% for multimodal screening. This implies that, in annual screening of a population with an incidence of 40 per 100,000, and if no cancers were missed, between 2.5 and 60 women would undergo surgery for every primary ovarian cancer detected. CONCLUSIONS: Ultrasound and multimodal screening can detect ovarian cancer in asymptomatic women, but there is currently no evidence on whether screening improves outcome for women in any risk group. On-going randomised controlled trials should establish the magnitude of any benefit of screening. The low prevalence of ovarian cancer in the population, and its rate of progression, may limit the potential cost-effectiveness of screening. PMID- 9853762 TI - Treatment with a gonadotrophin releasing hormone agonist before hysterectomy for leiomyomas: results of a multicentre, randomised controlled trial. AB - OBJECTIVES: To ascertain whether uterine shrinkage induced by a gonadotrophin releasing hormone agonist before hysterectomy for fibroids increases the possibility of a vaginal procedure. DESIGN: A multicentre, prospective, randomised, controlled study. PARTICIPANTS: One hundred and twenty-seven premenopausal women with a uterine volume of 12 to 16 gestational weeks. INTERVENTIONS: Twelve weeks of triptorelin depot treatment before hysterectomy or immediate surgery. MAIN OUTCOME MEASURES: Number of vaginal and abdominal hysterectomies, operating time, blood loss, degree of difficulty of the procedure, perioperative serum haemoglobin and haematocrit levels, hospital stay, and patients' overall satisfaction with treatment. RESULTS: After randomisation, four women withdrew from the study, leaving 60 women in the triptorelin arm and 63 in the immediate surgery arm. At baseline evaluation a vaginal hysterectomy was indicated in seven women allocated to pre-operative medical therapy (12%), and in 10 of those allocated to immediate surgery (16%). Clinical assessment after the 12-week GnRH agonist course showed that abdominal hysterectomy was no longer indicated in 25/53 women (47%) as a vaginal procedure appeared appropriate. Thus the overall rate of indication for a vaginal procedure in the pre-operative medical treatment arm was 32/60 cases (53%), with a between-group difference of 37% (95% CI, 26% to 51%; chi2(1) = 19.18, P < 0.0001; OR 6.06; 95% CI, 2.60 to 14.10). Pre- and post-operative serum haemoglobin and haematocrit levels were significantly higher in the GnRH agonist than in the immediate surgery arm. No appreciable difference was observed between the groups in the other intra- and post-operative variables, including patients' satisfaction. CONCLUSIONS: Pre-operative GnRH agonist therapy increased the rate of vaginal hysterectomy in selected women with fibroids and uterine volume of 12 to 16 gestational weeks. PMID- 9853763 TI - Perceptions of women on the impact of menorrhagia on their health using multi attribute utility assessment. AB - OBJECTIVE: To assess patient preferences regarding the treatment of menorrhagia using the multi-attribute utility methodologies, to produce a clinically applicable scale, and to assess health outcomes following treatment of menorrhagia. METHODS: Women referred to the gynaecology department for the treatment of menorrhagia were interviewed regarding the effects of menorrhagia on different aspects of their life. Their concerns were categorised into main components of health (domains). The relative importance of each domain was rated by the women using importance points which were distributed to represent the perceived importance of each domain. A series of statements (intra-domain statements) was developed for each domain, which described various possible conditions of that component of health. These were also rated using a one metre visual analogue scale with numerical anchor points at zero (worst) and 100 (best). RESULTS: The components of health considered most important were, in order of impact, family life, followed by physical health, work life, psychological health, practical difficulties and social life. The scores for the intra-domain statements were combined into a scale to allow the calculation of a final health state utility for a particular outcome based upon the statements the patient chooses within each domain. DISCUSSION: In planning treatment for menorrhagia clinicians can assess a woman's current perception of their health, using a simple to administer clinical scale. PMID- 9853764 TI - A systematic review comparing continuity of midwifery care with standard maternity services. AB - OBJECTIVE: To review randomised controlled trials of alternative maternity services characterised by continuity of midwifery care. METHODS: A systematic review of randomised controlled trials, analysed on an intention to treat basis, in which the study intervention was characterised by a midwife or small group of midwives providing care from early pregnancy to the postnatal period (defined as that provided on the postnatal ward); and the controls by standard maternity care as practised in the place where the trial was conducted. The seven trials identified included 9148 women. Main outcome measures were interventions during labour, maternal outcomes and infant outcomes. RESULTS: The alternative models with continuity of midwifery care were associated with less use of obstetric interventions during labour (eg, induction, augmentation of labour, electronic fetal monitoring, obstetric analgesia, instrumental vaginal delivery and episiotomy). However, the caesarean section rate did not differ statistically between the trial groups (OR 0.91; 95% CI 0.78 to 1.05). The lower episiotomy rate in the alternative models of care (OR 0.69; 95% CI 0.61 to 0.77) was associated with a significantly higher rate of perineal tears in the pooled alternative groups (OR 1.15; 95% CI 1.05 to 1.26). The percentage of intact perineums was very similar for the two groups (OR 1.11; 95% CI 1.00 to 1.24). There was no maternal death, and rates of maternal complications based on unpooled estimates did not show any statistically significant differences. The proportion of babies with an Apgar score < 7 at five minutes after the birth was approximately the same in the pooled alternative groups as in the control groups (OR 1.13 95% CI 0.69 to 1.84). Admission to intensive care or special care baby unit was similar (OR 0.86; 95% CI 0.71 to 1.04). The difference in perinatal deaths was bordering on statistical significance (OR 1.60; 95% CI 0.99 to 2.59). CONCLUSION: Continuity of midwifery care is associated with lower intervention rates than standard maternity care. No statistically significant differences were observed in maternal and infant outcomes. However, more research is necessary to make definite conclusions about safety, for the infant as well as for the mother. This review illustrates the variation in the different models of alternative and standard maternity care, and thus the problems associated with pooling data from different trials. PMID- 9853765 TI - A prospective intervention study of stopping smoking in pregnancy in a routine antenatal care setting. AB - OBJECTIVES: To evaluate the effect of specific education of midwives on stopping smoking in pregnant women and to determine the effect of this programme on pregnancy outcome. DESIGN: A prospective intervention study. SETTING: Department of Gynaecology and Obstetrics, Aarhus University Hospital, Denmark. POPULATION AND METHODS: A group of midwives (9 out of 54) was taught the effects of smoking during pregnancy and instructed in various methods of educating women to stop smoking. All pregnant women who attended antenatal care from October 1994 to September 1995 were either in the intervention group (n = 527) or the control group (n = 2629). Pregnant women attending the specifically educated midwives were considered to be the intervention group. They were given individual advice about stopping smoking at the first antenatal visit at about 16 weeks of gestation and a leaflet on smoking and pregnancy. RESULTS: At the first antenatal visit at about 16 weeks of gestation, 22% of the pregnant women smoked. Between the first visit and the routine visit at 30 weeks of gestation 51 (2%) stopped smoking and 56 (2%) started smoking. No differences were found between the intervention group and the control group in the rate of stopping smoking, validated by cotinine measurements. Mean birthweight, mean gestational age and the proportion of preterm birth in the two groups were similar. CONCLUSION: Education of midwives and integration of advice about smoking cessation at a low cost in routine antenatal care failed to affect smoking habits among pregnant women. PMID- 9853767 TI - Prevalence of heart disease diagnosed de novo in pregnancy in a West London population. AB - OBJECTIVES: To determine the prevalence of heart disease diagnosed de novo in pregnancy in a West London population and to re-examine the current role of routine cardiovascular examination in antenatal care in the UK. DESIGN: Retrospective study. SETTING: Obstetric medical clinics at Queen Charlotte's and Chelsea Hospital, University College Hospital and Northwick Park Hospital. POPULATION: Three hundred and twenty women referred for cardiac evaluation during pregnancy. RESULTS: The majority of the 139 women referred specifically for evaluation of murmurs during pregnancy were found to have physiological murmurs (97%). Only four women (3%) were found to have significant cardiac lesions de novo in their pregnancy. Three of these four women were immigrants who had no previous history of heart disease. The only woman from the UK was already known to have a murmur from childhood. CONCLUSIONS: Our study shows that heart disease diagnosed de novo in pregnancy in a West London population is an uncommon problem with low prevalence. It also appears to be a problem seen mainly in the immigrant population. The results emphasise the importance of taking a thorough medical history in all pregnant women. However, our results if they are confirmed, would suggest that only immigrants and those with significant symptoms or a known history of heart murmur or heart disease need undergo cardiovascular examination during pregnancy. These findings need to be confirmed in a larger group in other parts of the UK before further recommendations on selective cardiovascular examination can be made. This will have significant implications for midwifery led care. PMID- 9853766 TI - Does gestational hypertension become pre-eclampsia? AB - OBJECTIVE: To determine the likelihood of progression from gestational hypertension (GH) to pre-eclampsia (PE) in hypertensive pregnant women and whether this change can be identified in advance by available clinical and laboratory measures. DESIGN: A retrospective analysis and a prospective study. SETTING: St George Hospital, Sydney, a teaching hospital of the University of New South Wales delivering 2500 women per year. POPULATION: Eight hundred and forty five women with new hypertension in the second half of pregnancy, managed by a uniform protocol (661 in the retrospective analysis, 184 in the prospective study). METHODS: Clinical and laboratory data at initial presentation were compared among women with GH who developed PE and those who remained with a diagnosis of GH until delivery. Data predictive for progression from GH to PE were analysed by logistic regression analysis. MAIN OUTCOME MEASURED: Progression from GH to PE. RESULTS: In the retrospective analysis, 416 women initially presented as having GH and 62 (15%) progressed to PE. In the prospective study, 112 women initially presented with GH and 29 (26%) progressed to PE, giving an overall progression of 17%. In both studies, women who developed PE from GH presented earlier than those who remained with GH until delivery. In multiple logistic regression analyses prior miscarriage and early gestation at presentation were associated with increased likelihood of progressing from GH to PE. CONCLUSION: Approximately 15-25% of women initially diagnosed with GH will develop PE and this is more likely with earlier presentation or if the woman has had a prior miscarriage. Women with gestational hypertension diagnosed after 36 weeks of gestation have only about 10% risk of developing PE. These data should help stratify the risks of mildly hypertensive pregnant women being managed as outpatients in their third trimester. PMID- 9853768 TI - Potentially avoidable perinatal deaths in Denmark, Sweden and Lithuania as classified by the Nordic-Baltic classification. AB - OBJECTIVES: To analyse which subgroups of perinatal deaths contribute most to the perinatal mortality rate in Lithuania compared with Denmark/Sweden. DESIGN: Comparison of all perinatal deaths in Denmark/Sweden 1991 and Lithuania 1993-1994 by the common Nordic-Baltic perinatal death classification, based on information from the medical records. RESULTS: The doubled perinatal mortality in Lithuania compared with Denmark/Sweden is mainly explained by a threefold increase of intrapartum and two- to fivefold increase in neonatal deaths of nonmalformed infants. The higher rate of malformed infants is partly explained by a four times higher mortality from neural tube defects. CONCLUSIONS: The use of a common perinatal death classification has identified categories mainly responsible for the higher rate of perinatal mortality, but further investigation of potentially avoidable factors requires further study. PMID- 9853769 TI - Circulating markers of oxidative stress are raised in normal pregnancy and pre eclampsia. AB - OBJECTIVE: To determine whether circulating markers of oxidative stress are elevated in pre-eclampsia when appropriate precautions are taken to prevent in vitro oxidation DESIGN: A prospective study. SETTING: Nuffield Department of Obstetrics and Gynaecology, Oxford and The William Harvey Institute, London. SAMPLE: Three groups of women: those with pre-eclampsia (n = 19), control pregnant women (n = 19) matched for gestation, age and parity and a group of non pregnant individuals of reproductive age (n = 7). METHODS: Citrated plasma was stored at -80 degrees C with 20 micromol beta hydroxytoluene to prevent auto oxidation. Plasma samples were assayed for levels of 8 epi-prostaglandin F2alpha, lipid hydroperoxides, malondialdehyde and also the lipid soluble antioxidant vitamin E. RESULTS: There were no differences in 8 epi-prostaglandin F2alpha, lipid peroxide or malondialdehyde levels between the groups of women with pre eclampsia and those acting as pregnant controls. However, lipid hydroperoxides and malondialdehyde were significantly raised in both pre-eclampsia and normal pregnancy, compared with nonpregnant women. Vitamin E levels were similar in women with pre-eclampsia and those with a normal pregnancy, but in both groups levels were significantly higher than in nonpregnant women. CONCLUSION: Circulating markers of oxidative stress are raised in normal pregnancy and pre eclampsia. PMID- 9853770 TI - Intra-amniotic inflammation in human gastroschisis: possible aetiology of postnatal bowel dysfunction. AB - OBJECTIVE: To assess amniotic fluid for evidence of an inflammatory exudate in association with fetal gastroschisis. SETTING: University College Hospital, London and Institute of Child Health, London. SAMPLE: Samples of amniotic fluid in the third trimester from pregnant women with a diagnosis of fetal gastroschisis (n = 10) and from a control group (n = 10) with a normal fetus. METHODS: Cytological analysis of the fluid was performed. Flow cytometry was performed on the amniotic fluid using antibodies for the myeloid cell antigen CD15, the leucocyte beta integrin CD11b/CD18 and CD3, CD19, CD56 and CD25. Tumour necrosis factor alpha and interleukin-8 levels were assayed in the amniotic fluid. RESULTS: An acute inflammatory exudate, composed predominantly of neutrophil polymorphs and mononuclear cells, was found in the amniotic fluid in fetal gastroschisis but not in control cases. When amniotic fluid samples from cases of fetal gastroschisis were stained with CD15, analysis by flow cytometry showed a clear positive population. This CD15 population showed markedly elevated levels of CD11b. No distinct population of CD15 positive cells was seen in amniotic fluid samples examined from the control group. No staining was seen with antibodies to CD3, CD19, CD56 or CD25 in amniotic fluid obtained from either group. There was no significant difference between tumour necrosis factor alpha levels measured in the amniotic fluid of cases of fetal gastroschisis (median 102 pg/mL; range 20-340) and those of the control group (140 pg/mL; range 20-548) (P = 0.1). The levels of interleukin-8 were markedly elevated in the amniotic fluid of cases of fetal gastroschisis (median 6320 pg/mL; range 4732-13,800) compared with the control group (median 1738 pg/mL; range 623 2861;) (P < 0.01). CONCLUSION: Human fetal gastroschisis is associated with an inflammatory exudate in the amniotic fluid which may have implications for postnatal bowel function. PMID- 9853771 TI - A randomised placebo-controlled trial of ursodeoxycholic acid and S adenosylmethionine in the treatment of intrahepatic cholestasis of pregnancy. AB - A randomised placebo controlled study was performed in 32 women with intrahepatic cholestasis of pregnancy. The population was divided into four groups: for 20 days each group was treated only with ursodeoxycholic acid, or S adenosylmethionine, or a combination of both drugs, or a placebo (vitamin). Itching, standard liver function tests and serum total bile acids were measured before, during, and after treatment. Itching improved in all the women as well as the biochemical abnormalities. No side-effects in the mother or in the infant were recorded during and after therapy. A combination of ursodeoxycholic acid and S-adenosylmethionine is more effective than placebo and than either drug alone. PMID- 9853772 TI - Liver function following pregnancy complicated by the HELLP syndrome. AB - Serum levels of aminotransferases, lactate dehydrogenase, gammaglutamyl transferase, alkaline phosphatase, albumin and conjugated bilirubin, measured in 54 women at a median of 31 months (range 3-101) after pregnancies complicated by the HELLP syndrome, were not elevated. Total bilirubin levels, however, were elevated in 20% of these women; this represents a significant difference from the prevalence in 151 women with a previous normal pregnancy (chi2 = 12.23, P < 0.001), or in the normal female population (chi2 = 22.34, P < 0.00001). This raises the possibility that a dysfunction of the bilirubin-conjugating mechanism represents a risk factor for the development of the HELLP syndrome. PMID- 9853773 TI - Prevalence of faecal incontinence among women with urinary incontinence. AB - Four hundred and sixty-five women attending a urodynamic clinic were interviewed, and completed a detailed bowel questionnaire, about their urinary and bowel symptoms. All the women underwent video-cystourethrography with pressure and flow studies. The reported incidence of faecal incontinence was 15.3% (n = 71) on direct questioning and 26% (n = 121) on the postal questionnaire. Faecal incontinence was more common in women with a urodynamic diagnosis of detrusor instability (30% (n = 26)) than among women diagnosed as having genuine stress incontinence (21% (n = 38)). Denervation and myogenic injuries sustained during childbirth have been suggested as a common cause for genuine stress and faecal incontinence, but there may be an alternative mechanism to explain why women with detrusor instability suffer from faecal incontinence. PMID- 9853774 TI - The weak cervix: failing to keep the baby in or infection out? PMID- 9853775 TI - Laparoscopic repair of a vesicovaginal fistula using an omental J flap. PMID- 9853777 TI - Systematic reviews: insufficient evidence on which to base medicine. PMID- 9853778 TI - Systematic reviews: insufficient evidence on which to base medicine. PMID- 9853779 TI - Analysis of birthweight and gestational age in antepartum stillbirths. PMID- 9853780 TI - Obstetrics and gynaecology, and women's health. PMID- 9853781 TI - Laparoscopic colposuspension. PMID- 9853782 TI - Chronic pelvic pain. PMID- 9853783 TI - The collaborative randomised aminoinfusion for meconium project (CRAMP): 1. South Africa and 2. Zimbabwe. PMID- 9853784 TI - Impact of metronidazole therapy on preterm birth in women with bacterial vaginosis flora. PMID- 9853785 TI - Schizophrenia research: a biennium of progress. Proceedings from the Sixth International Congress on Schizophrenia Research. Colorado Springs, CO, April 12 16, 1997. AB - The Sixth International Congress on Schizophrenia Research (ICOSR) took place in Colorado Springs, Colorado, April 12-16, 1997, where over 1,000 scientists presented and listened to the latest developments in the search for the cause and treatment of schizophrenia. The ICOSR is sponsored by Maryland Psychiatric Research Center, Case Western Reserve University, and the William K. Warren Foundation. The National Institute of Mental Health and several pharmaceutical companies contributed generously to the meeting. The ICOSR is co-organized by Dr. Carol A. Tamminga, Maryland Psychiatric Research Center, University of Maryland at Baltimore, and Dr. S. Charles Schulz, Case Western Reserve University, Cleveland, Ohio. The William K. Warren Research Award is given to a senior investigator, who has made outstanding contributions to our understanding of schizophrenia. The fifth William K. Warren Research Award was presented to Dr. Philip S. Holzman in recognition of his contributions to the identification of eye-tracking abnormalities as a potential phenotypic marker of the illness and also in recognition of his work as a lifelong mentor for schizophrenia researchers. The ICOSR Young Investigator Awards are presented to junior investigators who have demonstrated the potential to make significant contributions to research on schizophrenia. These awards promote scientific development by enabling these young researchers to attend the meeting. There were 30 Young Investigator Award winners. The ICOSR meeting is organized into four sessions: (1) a morning plenary session; (2) a plenary lecture; (3) a poster session; and (4) concurrent afternoon oral sessions. The morning plenary sessions are comprised of a set of 30-minute lectures, which provide an overview of a particular topic area relevant to schizophrenia research. The plenary lecture is an invited lecture on a basic topic related to current research efforts in schizophrenia. The poster sessions provide a forum for the presentation of prepublication reports of basic and clinical science projects. The afternoon sessions are a collection of approximately 10 focused presentations on current research projects related to a specific topic area. The purpose of this report is to provide an account of the proceedings from the plenary and afternoon oral sessions. PMID- 9853786 TI - Health care expenditure on schizophrenia patients in Belgium. AB - Schizophrenia generates a great deal of cost and burden. The aim of this study was to assess the direct costs for schizophrenia patients receiving standard treatment in different settings in Belgium. Costs were calculated for patients and for the Belgian insurance system. Data from Belgium's largest sickness fund were used to estimate health expenditures for all schizophrenia patients in Belgium. The mean direct treatment cost was $12,050 per patient per year, or $304 million for all schizophrenia patients per year. This cost constitutes 1.9 percent of the Belgian Government's total health expenditure. Government expenditure per schizophrenia patient is 10 times that of an average citizen. PMID- 9853787 TI - Changes in the risk of schizophrenia in Scotland: is there an environmental factor? AB - Various studies have suggested that there may be an environmental factor in schizophrenia acting before or at birth but with delayed effects. Evidence that the risk of developing schizophrenia varies randomly with the year of birth would help confirm the existence of such a factor. Data from the Scottish Health Service Information and Statistics Division, comprising all first admissions for ICD-9 schizophrenia in the years 1963-91, were used to estimate the lifetime risk of developing schizophrenia for each year of birth from 1900 to 1969. In the period after 1928 the lifetime risk steadily declined. The rate of decline was greater in females. The random fluctuations in the risks in females did not change systematically, but there was a significant decline in the variability of these fluctuations in males. These random fluctuations suggest a possible role for randomly varying environmental influences around the time of birth. Our findings suggest a possible diminution in the role of such environmental influences in schizophrenia among males in Scotland in the years 1929 to 1969. PMID- 9853788 TI - The Russian concept of schizophrenia: a review of the literature. AB - The focus of this article is a comprehensive review of the Russian-Soviet conceptualization of schizophrenia, which can be understood only in the broader historical and cultural context of Russian-Soviet psychiatry. Because of multiple barriers and the political abuse of psychiatry in the former Soviet Union, international psychiatric literature has lacked unbiased data about the scientific merit and historical logic of the Russian-Soviet concept of schizophrenia. This article represents an attempt to examine phenomenology, nosology, and some biological theories of schizophrenia developed in the former U.S.S.R. from historical and scientific points of view and to compare them to the Western theories. The article also addresses historical and cultural antecedents of the abuse of psychiatry. The author suggests that the lack of a democratic tradition in Russia, a totalitarian regime, and oppression and "extermination" of the best psychiatrists during the 1930-50 period prepared the ground for the abuse of psychiatry and Russian-Soviet concept of schizophrenia. Perspectives on the potential changes in the Russian concept of schizophrenia in changing historical conditions are discussed. PMID- 9853789 TI - The quality of life of people with schizophrenia in Boulder, Colorado, and Bologna, Italy. AB - The aim of the study was to compare the quality of life (QOL) and needs of people with schizophrenia in comprehensive treatment systems in two countries. One hundred people with schizophrenia and schizoaffective disorder were randomly selected from the caseload of a community mental health center in Boulder, Colorado, and 70 were similarly selected from public psychiatric treatment services in and around Bologna, Italy. Subjects were interviewed with QOL and needs assessment instruments and rated with the Brief Psychiatric Rating Scale. Objective QOL measures favored Bologna subjects over Boulder subjects, particularly with respect to employment, accommodation, and family life. In a factor analysis, objective QOL variables sorted separately from subjective satisfaction ratings, suggesting that they measure different underlying constructs. Patient needs in both Boulder and Bologna samples were primarily psychological and social rather than basic survival issues. Boulder subjects were more likely to report accommodation needs. Many apparent QOL advantages for Bologna subjects could be attributed to the greater frequency with which the Italian patients lived with family of origin. Living with family also appeared to confer practical benefits in meeting needs. Objective QOL measures discriminated between patient populations better than subjective ratings of satisfaction and well-being. PMID- 9853790 TI - Role of psychosocial treatments in management of schizophrenia: a meta-analytic review of controlled outcome studies. AB - This meta-analytic review sought to answer questions concerning the role of psychosocial treatments in the comprehensive management of patients with schizophrenia. The review focused on the effects of combining psychosocial treatment with somatic treatment. Findings demonstrated the additive and supplementary effects of psychosocial treatments and the durability of these effects. Patients with more chronic illness appeared to be more responsive to psychosocial treatments, as were patients in studies conducted in non-Western countries. Among the Western countries, studies from Scandinavian countries reported the least effectiveness for psychosocial treatments. There was some evidence for differential effect of psychosocial treatments on different dimensions of illness as the measures of disorganized behavior and employment showed little difference in treated and control groups. There was also some evidence for differences between different modalities of treatment as group treatments produced smaller effects. Implications for practice and future research are discussed. PMID- 9853792 TI - Family caregiving in schizophrenia: domains and distress. AB - This article focuses on (1) the dimensionality of the caregiving concept; (2) the relation between the identified caregiving dimensions and characteristics of the patient, the caregiver, and their relationship; and (3) the relation between caregiving dimensions and caregiver distress. Findings are based on data from 480 members of the Dutch family organization for patients with schizophrenia/chronic psychosis who completed (1) the Involvement Evaluation Questionnaire (IEQ), which assesses general information (e.g., household characteristics), caregiving, help seeking, coping and distress, and (2) a questionnaire comprising questions on onset and course of the patient's disorder and symptoms characteristic of schizophrenic disorders. Four caregiving domains were found: tension, supervision, worrying, and urging. These domains were strongly related to the patient's symptomatology, contact between the relative and the patient's mental health professional, and the number of hours of mutual contact between the patient and the relative. The connection between patient, caregiver, and relationship variables and the caregivers' distress could be explained substantially by the overall caregiving score. Our findings suggest that caregiving tasks and problems may be diminished and related distress lowered by reducing the patient's symptomatology, increasing relatives' coping capacities, and decreasing the number of contact hours. If distress is reduced, relatives may use less psychotropic medication and may visit their general practitioner less often. PMID- 9853791 TI - Review of integrated mental health and substance abuse treatment for patients with dual disorders. AB - Patients with severe mental disorders such as schizophrenia and co-occurring substance use disorders traditionally received treatments for their two disorders from two different sets of clinicians in parallel treatment systems. Dissatisfaction with this clinical tradition led to the development of integrated treatment models in which the same clinicians or teams of clinicians provide substance abuse treatment and mental health treatment in a coordinated fashion. We reviewed 36 research studies on the effectiveness of integrated treatment for dually diagnosed patients. Studies of adding dual-disorders groups to traditional services, studies of intensive integrated treatments in controlled settings, and studies of demonstration projects have thus far yielded disappointing results. On the other hand, 10 recent studies of comprehensive, integrated outpatient treatment programs provide encouraging evidence of the programs' potential to engage dually diagnosed patients in services and to help them reduce substance abuse and attain remission. Outcomes related to hospital use, psychiatric symptoms, and other domains are less consistent. Several program features appear to be associated with effectiveness: assertive outreach, case management, and a longitudinal, stage-wise, motivational approach to substance abuse treatment. Given the magnitude and severity of the problem of dual disorders, more controlled research on integrated treatment is needed. PMID- 9853793 TI - Relationship between empathic family attitude and relapse in schizophrenia: a 2 year followup prospective study. AB - Empathy plays a central role in social relationships, and lack of empathy has been suggested as part of expressed emotion in the relatives of patients with schizophrenia. The aim of this research is to measure empathy in the relatives of schizophrenia patients and to establish the relationship between lack of empathy and relapse. Eighty schizophrenia patients were followed up in a 2-year prospective cohort study. Relatives' empathy, defined as the ability to perceive the patient's mood state, was measured at the beginning of the study with a questionnaire given after a 10-minute interaction between the patient and his or her relative. Several other attitudinal, clinical, and social variables were also measured. A significant relationship was found between poor empathic attitude and relapse. Lack of treatment compliance, negative symptoms, unemployment, and poor premorbid adjustment were also associated with relapse. In a multivariate analysis, the association between poor empathic attitude and relapse was maintained. Statistical control of the relatives' critical attitude showed that each kind of attitude predicts relapse independently. PMID- 9853794 TI - Suicidal ideation and suicide attempts in recent-onset schizophrenia. AB - The objective of this study was to predict suicidality in people with schizophrenia. Ninety-six patients with recent-onset schizophrenia were rated every 2 weeks for 1 year to examine (1) the temporal course of suicidal ideation and suicide attempts and (2) the extent to which anxiety, depression, and mild suicidal ideation were followed by significant suicidal ideation or a suicide attempt. The severity of suicidality changed rapidly. Low levels of suicidal ideation increased the risk for significant suicidal ideation or a suicide attempt during the subsequent 3 months. Depression was moderately correlated with concurrent suicidality, but not independently associated with future suicidality. Therefore, low levels of suicidal ideation may predict future suicidal ideation or behavior better than depressed mood in individuals with schizophrenia. PMID- 9853795 TI - Verbal learning and memory in schizotypal personality disorder. AB - The investigation of cognitive deficits in patients with schizotypal personality disorder (SPD) is important both to establish commonalities between SPD and schizophrenia and to clarify the significance of these cognitive deficits for schizophrenic disorders. The purpose of this study was to examine verbal learning and memory with the California Verbal Learning Test (CVLT) in a group of patients with SPD (n=24) and a group of patients with personality disorders other than SPD (OPD; n=25). The results indicated that SPD patients learned significantly fewer words with practice on the CVLT than OPD patients (F=4.32, df=1,47, p < 0.05), and their rate of learning was reduced relative to normative standards. These findings suggest that SPD patients have a deficit in verbal learning that is similar to, although not as severe as, the impairments seen in schizophrenia. PMID- 9853796 TI - Auditory laterality and selective attention: normal performance in patients with early-onset schizophrenia. AB - In this study, auditory laterality and selective attention were examined in patients with early-onset schizophrenia using a dichotic listening (DL) test. Deficient performance on this test has repeatedly been found in adult patients with chronic schizophrenia, indicating abnormalities in left hemisphere function. The hypothesis in the present study was that subjects with early-onset schizophrenia manifest deficits in DL test performance similar to adult chronic patients. A group of 19 patients with early-onset schizophrenia were compared with a group of 20 adolescents with attention-deficit hyperactivity disorder and a group of 30 normal adolescents. Results indicated no significant differences between the three groups on any of the measures. Alternative hypotheses are put forth to explain the findings, among them that deficits in DL performance may be secondary to long-time illness and/or drug treatment, and that these deficits may become apparent only after interaction with maturational neurodevelopmental changes during adolescence. PMID- 9853797 TI - First person account: psychosocial implications of stigma caused by misdiagnosis. AB - The article that follows is part of the Schizophrenia Bulletin's ongoing First Person Account series. We hope that mental health professionals--the Bulletin's primary audience--will take this opportunity to learn about the issues and difficulties confronted by consumers of mental health care. In addition, we hope that these accounts will give patients and families a better sense of not being alone in confronting the problems that can be anticipated by persons with serious emotional difficulties. We welcome other contributions from patients, ex patients, or family members. Our major editorial requirement is that such contributions be clearly written and organized, and that a novel or unique aspect of schizophrenia be described, with special emphasis on points that will be important for professionals. Clinicians who see articulate patients, with experiences they believe should be shared, might encourage these patients to submit their articles to First Person Accounts, Office of Scientific Information, NIMH, 5600 Fishers Lane, Rm. 10-85, Rockville, MD 20857. PMID- 9853798 TI - Kinetic study of apo B100 containing lipoprotein metabolism using amino acids labeled with stable isotopes: methodological aspects. AB - Kinetic disturbances of lipoprotein metabolism are important to know for a better understanding of lipid diseases or effects of drugs. These kinetic aspects were previously studied with radioactive tracers. The ethical concerns related to these tracers can be now overcome at a reasonable cost with the new development of small bench top mass spectrometers and the increased production of stable isotope tracers. In this review, we will discuss some methodological aspects related to stable isotope tracers and the analysis of the data with non compartmental or compartmental models. PMID- 9853799 TI - Activated immune system in patients with Huntington's disease. AB - Abnormalities of immune system compartments were determined in 12 patients with Huntington's disease (eight males, four females; age 42.4+/-11.7 years) and 11 controls (7 males, 4 females; age 47.0+/-12.0). All patients were free from infectious diseases. Serum concentrations of a panel of serum soluble markers of immune activation were investigated, namely neopterin, 55-kDa-type soluble tumor necrosis factor receptor (sTNF-R), interleukin-2-receptor (sIL-2R), kynurenine, tryptophan, immunoglobulins (Ig) A, M and G as well as routine laboratory tests. Compared to controls, we found significantly higher serum levels of IgA (p<0.01), sTNF-R, sIL-2R, neopterin, and complement component C3 (all p<0.05), and serum tryptophan was decreased (p<0.001). Higher concentrations of circulating immune complexes, cardiolipin antibodies, IgM, neopterin and lower tryptophan were associated with loss of cognitive function as assessed by the mini-mental-test. Five patients died within 1 year after measurements were performed. In these patients IgM, circulating immune complexes and neopterin concentrations were higher compared to survivors and serum tryptophan was lower. The data indicate an activation of various immune system compartments in Huntington's disease and that systemic immunological alterations might be important in the course of the disease. PMID- 9853800 TI - Elevated group II phospholipase A2 mass concentration in serum and colonic mucosa in Crohn's disease. AB - Group II phospholipase A2 has been proposed to play an important role in the pathophysiology of inflammatory bowel diseases. This enzyme has also been linked to host defence mechanisms against bacteria. The current study aimed at measuring the mass concentrations of group II phospholipase A2 in serum and colonic mucosa of patients with Crohn's disease of different severity and of appropriate control patients without any inflammatory disease. The activity of the disease was determined by clinical factors (the simple index score) and endoscopic and histological scoring. The mass concentration of group II phospholipase A2 was measured by a time-resolved fluoroimmunoassay. The mass concentrations of group II phospholipase A2 in serum and colonic mucosa were significantly higher both in patients with active and inactive Crohn's disease when compared with controls. There was statistically significant difference in the mass concentration of group II phospholipase A2 in colonic mucosa but not in serum between inactive and active Crohn's disease. The current results indicate that the mass concentration of group II phospholipase A2 is increased in serum and colonic mucosa of patients with Crohn's disease and that the latter is associated with the degree of the inflammatory activity in the intestinal wall. These results support the idea that group II phospholipase A2 is involved in the local and generalised pathological processes of Crohn's disease. PMID- 9853801 TI - Determination of pH in human erythrocytes. Sources of systematic error. AB - The results of a simultaneous determination of the pH value in erythrocytes with the potentiometric measuring method and the 14C-labelled 5,5-dimethyl-2,4 oxazolidinedione (DMO) method showed a mean method difference of 0.026 pH units. The cause of this discrepancy was assumed to be matrix-inherent liquid-junction potentials in the potentiometric measurement. Taking these into account in the calculation leads to consistent values for the methods investigated. The DMO method proved to be free of systematic errors. Another indication of this is that its mean ratio of extracellular to intracellular H+ ion concentration (H+e/H+i) substantially agreed with the distribution ratios of other freely diffusible ions and their pH dependence. PMID- 9853802 TI - Rapid molecular diagnosis of erythropoietic protoporphyria among Swiss patients. AB - Erythropoietic protoporphyria (EPP) is an autosomal dominant inherited disorder with incomplete penetrance. It is caused by partial deficiency of ferrochelatase, the last enzyme in the heme biosynthetic pathway. Measurement of protoporphyrin concentrations in red cells and feces, although sufficient for diagnosis of symptomatic EPP patients, fails to detect asymptomatic gene carriers. We have developed a molecular diagnostic procedure for rapid and reliable screening of five known mutations in the ferrochelatase gene among Swiss EPP patients in a single denaturing gradient gel electrophoresis (DGGE) gel. PMID- 9853803 TI - Biochemical evaluation of oxidative stress in propylthiouracil treated hyperthyroid patients. Effects of vitamin C supplementation. AB - In this study the impact of vitamin C supplementation on oxidative damage as assessed by thiobarbituric acid reactive substances and markers of antioxidant status: namely Cu/Zn superoxide dismutase, glutathione peroxidase, glutathione reductase and glutathione were investigated in 24 hyperthyroid patients under propylthiouracil therapy (3x100 mg/day) for five days and in 15 healthy controls. Ascorbic acid (1000 mg/day) was given as a supplement for 1 month to both the patients and controls during the study period. Heparinised blood samples were taken at the beginning and the end of one month ascorbic acid supplementation. Comparison of the hyperthyroid patients with the controls revealed higher lipid peroxidation (p<0.001), higher Cu/Zn superoxide dismutase activity (p<0.001), higher glutathione level (p<0.001) and lower glutathione reductase activity (p<0.001). Vitamin C supplementation to hyperthyroid patients caused significant increases in glutathione concentration (p<0.001) and glutathione peroxidase activity (p<0.001), whereas there were significant decreases in glutathione reductase (p<0.001) and Cu/Zn superoxide dismutase activities (p<0.01). Thiobarbituric acid reactive substances and thiobarbituric acid reactive substances/glutathione ratio were significantly decreased (p<0.01). Vitamin C supplementation to euthyroid controls caused significant increases in glutathione concentration (p<0.001) and glutathione peroxidase and Cu/Zn superoxide dismutase activities (p<0.001), whereas there was a significant decrease in glutathione reductase (p<0.001). The thiobarbituric acid reactive substances/glutathione ratio was significantly decreased (p<0.05). Our findings reveal the potentiation of antioxidant status and a relief in oxidative stress in both propylthiouracil treated hyperthyroid patients and controls in response to vitamin C supplementation. PMID- 9853804 TI - Effect of aminoguanidine on erythrocyte lipid peroxidation and activities of antioxidant enzymes in experimental diabetes. AB - The effect of aminoguanidine (AG) on the malondialdehyde (MDA) concentration and activities of superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-Px) in erythrocytes of rats with streptozotocin-induced diabetes was studied. Induction of diabetes resulted in an increase of MDA concentration and decreases of SOD and catalase activities after 6 and 12 weeks. GSH-Px activity increased after 6 weeks and returned to control values after 12 weeks. AG administration did not affect body weight, blood glucose level and HbA1c content in diabetic rats but led to a decrease of MDA concentration and SOD and catalase activities after 12 weeks of treatment, with no significant effect after 6 weeks. AG attenuated the GSH-Px increase after 6 weeks but augmented the activity of this enzyme after 12 weeks. These results confirm the presence of oxidative stress in streptozotocin-induced experimental diabetes and point to the beneficial antioxidant effect of AG. PMID- 9853805 TI - Normalizing intermethod free triiodothyronine patient results: need for commutable materials. AB - The aim of this work was to check the suitability of control materials to normalize intermethod results for the measurement of free triiodothyronine in patient sera. In the main experiment, 108 patient sera and 11 commercially available control materials were assayed by a pair of methods. In a confirmatory experiment, two of the control materials and 142 patient sera were assayed with an alternative pair of methods. In the main experiment, the intermethod variability of 6/11 control materials differed significantly from that of patient sera (i. e. control materials were non-commutable). Recalculation of patient results using control materials as calibrators lowered the intermethod difference only if commutable materials were used. The confirmatory experiment demonstrated that the pattern of commutability changed if a different pair of methods was used. We conclude that in the case of free triiodothyronine the commutability of control materials should be tested if they are to be used to normalize patient results obtained by different methods. PMID- 9853806 TI - Evaluation of a predictive kinetic procedure for the enzymatic measurement of creatinine in serum. AB - We have evaluated the analytical performance of a predictive kinetic procedure for the enzymatic measurement of serum creatinine in comparison with equilibrium and kinetic fixed-time procedures. The procedure uses commercially available reagents and is based on calculating the total change in absorbance expected if the reaction were to proceed to equilibrium by fitting the initial part of the absorbance-time curve to a three-parameter function by non-linear regression. Total imprecision for different concentrations of creatinine ranges from 1.47 to 1.86% and is significantly better than that obtained for the fixed-time procedure. The results with the predictive procedure are less dependent on creatinine amidohydrolase activity in the reagent, pH of the reagent and temperature of analysis than are the results with the fixed-time procedure. All the procedures are interfered with by bilirubin and ascorbic acid to about the same extent. Recovery and linearity are quite acceptable, and the estimated accuracy is below 2.1%. However, in comparison with the equilibrium procedure, the predictive procedure tends to underestimate creatinine concentrations at values below 100 micromol/l, and the results obtained by the two procedures are not transferable. In conclusion, the predictive approach substantially improves the imprecision but not the specificity of the enzymatic assay of serum creatinine. PMID- 9853807 TI - Pre-evaluation and system optimization of the Elecsys thyroid electrochemiluminescence immunoassays. AB - We present the results of a pre-evaluation of the thyroid function test free thyroxine, free triiodothyronine and third generation TSH using the Elecsys electrochemiluminescence immunoassay system. A collaborative field study between the development center of the manufacturer and a clinical chemistry laboratory addressed the reliability and comparability of the new Elecsys assays to established methods under clinical laboratory conditions using samples from routine in vitro thyroid testing. Preliminary (reference) formulations of the reagents and several electrochemiluminescent pilot models were used for assay measurements, either in the company's research center or in the clinical setting. The new thyroid assays were compared with the respective Enzymun-Test assays, performed on the ES300 automated immunoassay analyzer. A WHO standard was used for standardization of TSH, whereas an equilibrium dialysis method was applied for free triiodothyronine. The free thyroxine assay was standardized against the Enzymun-Test free thyroxine assay, which had previously been calibrated against equilibrium dialysis. The aim of this field study was to support the optimization of the technology used for Elecsys in an early stage of development and thereby prepare the ground for the adaptation of the immunoassays to the final Elecsys 2010 random access analyzer. A subsequent multicenter evaluation demonstrated that the requirements of routine thyroid testing in terms of reliability were fulfilled by the system. PMID- 9853808 TI - Validation of an automated enzyme immunoassay for Interleukin-6 for routine clinical use. AB - Serum levels of Interleukin-6 (IL-6), a proinflammatory cytokine, are increased in early stages of inflammatory diseases such as infection and sepsis. Assay systems which permit its measurement within a few hours and as a single measurement have not been reported so far. We therefore evaluated a now commercially available automated method for IL-6 measurement on the Cobas Core immunological analyzer (Roche Diagnostic Systems) which enables single IL-6 measurement within about 1 hour. The automated assay correlates well with an established, manual microtiter plate assay (Biosource GmbH) which uses the same antibodies and reagents (r=0.98). Accuracy of the automated method was established by adding known amounts of IL-6 international reference preparation. Recovery of the international standard was in the range of 92104%. The automated assay had a precision of singletons below 6% and was linear up to 2800 pg/ml. This automated assay provides a suitable, convenient and time saving method for measurement of IL-6 serum levels in the routine clinical laboratory. PMID- 9853809 TI - First experience with the REMEDi HS urine benzodiazepine assay. AB - Ninety eight urine samples were analysed with an immunoassay benzodiazepine kit. A total of 68 urine specimens that were presumptively positive for benzodiazepines were evaluated by the REMEDi HS urine benzodiazepine assay (BIO RAD, Munich, Germany). Of this number, 53 (78%) specimens were found by REMEDi to contain one or more benzodiazepines or their metabolites, and 15 (22%) were found to be negative. From the discordant group of 15 samples, eight were found to be negative using conventional chromatographic procedures (HPLC or GC/MS), while seven contained one or more benzodiazepines or metabolites, each of which were below the individual cut-off level specified by the manufacturer. Additionally 30 urine specimens that were negative for benzodiazepines using immunoassay were also tested by REMEDi. Two samples were found to be positive. These results could not be confirmed by other chromatographic techniques. The REMEDi HS benzodiazepine assay can be a very useful complementary technique in the clinical/forensic toxicology laboratory, especially for the identification of the parent benzodiazepines administered. The assay provides a rapid result in emergency situations and is useful in confirmation of preliminary positive immunoassay results. PMID- 9853811 TI - Are the preanalytical factors underestimated in clinical studies? PMID- 9853810 TI - Rapid variations of the free prostate-specific antigen fraction in serum. PMID- 9853812 TI - Egg culture: the foundation. AB - Ralph Brinster began his classic work on egg culture more than 35 years ago. His interest in mammalian egg culture had developed, in part, as a consequence of his experiences with animal breeding and reproduction that he gained while growing up on a farm. Ralph decided early in his career that an in vitro approach to culturing eggs would provide a powerful tool with which to study the development of these cells. Beginning at the close of the 19th century, a number of investigators had performed in vitro studies on egg culture and the related area of egg transfer; however, the ability to recover and transplant eggs had reached a much higher level of perfection than had culture. Eggs of many species could be successfully transferred, but there was no reliable technique for egg culture. In 1963, Ralph reported a method for culturing eggs in microdrops of medium under oil (Brinster, 1963), which has become universally used. Two years later, he identified pyruvate as the central and essential energy source for early stages of mouse eggs (Brinster, 1965b). These two developments revolutionized in vitro studies of mammalian eggs and issued in an era of intense research activity concerning egg culture and egg manipulation. Effective formulations of culture media could now be developed to allow routine in vitro maintenance of eggs, and important parameters for these recipes were soon determined. It was quickly established that the requirement for pyruvate as an energy source exists at ovulation in many species and is already present in germ cells of the mouse fetus. The metabolic activity of the fertilized mouse egg was shown to be low and comparable to bone; however, four days later, at the blastocyst stage of development, the metabolic activity was comparable to brain. Thus, a foundation of understanding about the biology of early mammalian eggs was established between 1960 and 1970, and subsequent studies have broadened this understanding. However, the greatest impact of a simple, reliable egg culture method has been to provide the ability to perform complicated manipulative procedures on preimplantation stages of mammalian embryos. In no area has this been more important than in development of transgenic animals. All methods for generating germ line genetic modifications rely on the ability to maintain and manipulate eggs and early developmental stages in vitro without loss of developmental competence. The importance of efficient egg culture to manipulation and transgenesis is fundamental and enabling. PMID- 9853813 TI - The coming of age of the transgenic era. PMID- 9853814 TI - Transgenic mice--the early days. PMID- 9853815 TI - Germ cells and germ cell transplantation. AB - The germ cell lineage in mice is established about a week after fertilization, in a group of cells that have left the epiblast and moved to an extraembryonic site. They migrate back into the embryo, along the hind gut and into the gonads. Germ cells in male and female embryos then pursue different pathways: in the testis the germ cells cease proliferating and enter mitotic arrest, while germ cells in the ovary, like those in male embryos that remain outside the gonads, enter meiotic prophase. Studies on explanted germ cells suggest that all germ cells may enter meiosis at a certain stage of their development, unless prevented from doing so by some inhibitory influence of the testis. Germ cells during the migratory stage can be cultured, but do not enter meiosis unless embedded in somatic tissue. Addition of certain growth factors and cytokines to the culture medium allows germ cells to proliferate indefinitely in vitro: Like embryonic stem cells, these immortalized EG (embryonic germ) cells will colonize all cell lineages if introduced into a blastocyst. After birth, germ cells undergo gametogenesis; oogenesis in the female, spermatogenesis in the male. Brinster and his colleagues have shown that spermatogonial stem cells injected into a germ cell depleted testis will repopulate the seminiferous tubules and undergo spermatogenesis, giving rise to functional spermatozoa. Stem cells from frozen testicular tissue are still capable of giving rise to spermatogenesis in a host testis. Rat testicular tissue can undergo spermatogenesis in a mouse testis, to form morphologically normal rat spermatozoa, even though the Sertoli cells that support them are of endogenous mouse origin. These findings are of fundamental importance for our understanding of spermatogenesis and the interactions between germ cells and Sertoli cells; but they also have significant practical implications, in relation to both agricultural practice and clinical treatment of infertility. PMID- 9853816 TI - Embryo culture, stem cells and experimental modification of the embryonic genome. An interview with Professor Ralph Brinster.. Interview by Juan Arechaga. PMID- 9853817 TI - Reflections on the culture of the preimplantation embryo. AB - There has been a considerable improvement in the media available for the culture of preimplantation mouse embryos during the 40 years since mouse embryos were first cultured and successfully transferred to uterine foster mothers. Two new media, KSOM and mMTF, are becoming more commonly used. The history of the development of these media, including recent work on KSOM and mMTF, is reviewed. A major artefact in the earlier work was the two-cell block. The causes of the two-cell block and the methods by which it has been overcome are reviewed. It is concluded that even the best available media inevitably cause imbalances in the environment in which the embryos are forced to develop, because they consist of only a small subset of the compounds present in the natural environments. As a result, the embryos must adapt to these abnormal conditions if they are to survive. The implications of these conclusions on the choice of media for specific purposes are discussed. PMID- 9853818 TI - Culture of human preimplantation embryos to the blastocyst stage: a comparison of 3 media. AB - Following culture for 2 days in Earle's balanced salt solution (EBSS), human embryos which remained after transfer were cultured in one of 3 media for 4 days, from the 2- to 4-cell stage to the blastocyst stage. Sibling embryos were divided equally between treatments. Throughout the 4 day culture period, embryos were assessed for morphology and development, as well as uptake and production of energy substrates. Cell numbers in the inner cell mass and trophectoderm were determined for embryos which reached the blastocyst stage. No significant effect was observed in the extent or rate of development to the 8-cell, morula or blastocyst stage between treatments. Uptake of pyruvate was related to concentration in the medium and no differences in glucose uptake were observed between media. Endogenous energy metabolism, as measured by lactate production, was significantly higher in Ham's F12 than in EBSS from day 3.5 onwards. Blastocyst cell numbers were also increased; 79.6+/-7.7 in Ham's F12 (n=17) and 57.8+/-5.2 in EBSS (n=19), p<0.05. Of the embryos which reached the blastocyst stage by day 5, 36% (14/25) had degenerated by day 6 in EBSS compared to only 19% (5/27) in Ham's F12 (p=0.06). Slightly higher rates of embryo survival between day 5 and 6 in Ham's F12 may account for the observed increase in blastocyst cell number. The results do not suggest that improved embryo development can be obtained using human tubal fluid or Ham's F12, in preference to EBSS during early cleavage stages, but the use of Ham's F12 may improve embryo survival at later stages of development. PMID- 9853819 TI - Postimplantation mouse development: whole embryo culture and micro-manipulation. AB - Methods for growing whole mouse embryos in vitro have been greatly improved in the last two decades. The present technology enables embryos to develop remarkably close to what can be achieved in vivo from the pre-gastrula to the early organogenesis stages. The ability to grow whole embryos for a substantial period outside the uterine environment offers a unique opportunity to observe the progression of development, and permits the performance of direct manipulation on the embryo. Experiments combining whole embryo culture and micro-manipulation have led to the discovery of new information on lineage differentiation, tissue interaction and morphogenetic mechanisms that are associated with the establishment of the fetal body plan. PMID- 9853820 TI - Mouse chimaeras revisited: recollections and reflections. AB - This very personal and subjective article briefly describes the evolution of techniques used for obtaining mouse chimaeras from various sources (embryos, EC, ES and EG cells), summarizes studies on inter-specific chimaeras, mentions some of other applications ('rescuing' chimaeras), presents the contribution of Ralph Brinster to this area and tries to estimate whether the expectations I expressed in 1961 as to the usefulness of making and studying chimaeras turned out to be correct. Tribute is paid mainly to those, who as the first, contributed to various aspects of these studies. PMID- 9853821 TI - Neurulation in amniote vertebrates: a novel view deduced from the use of quail chick chimeras. AB - Two apparently different mechanisms successively contribute to the formation of the neural tube in the avian embryo: bending of the neural plate during the primary neurulation in the cephalo-cervico-thoracic region and cavitation of the medullary cord during the secondary neurulation in the lumbo-sacral region. During both these processes, gastrulation continues by the caudal regression of Hensen's node--also called cordoneural hinge in the secondary neurulation. Labeling of Hensen's node or cordoneural hinge by the quail chick marker system revealed that this structure, which is the equivalent of the dorsal blastoporal lip of the Amphibian embryo, i.e., of the Spemann's organizer, gives rise to the midline cells of the three germ layers: the floor plate of the neural tube, the notocord and the dorsal cells of the intestinal endoderm. Caudally to the organizer, both in primary and secondary neurulation, the presumptive territory of the alar plates of the future neural tube overlies the precursors of the paraxial mesoderm. Regression of Hensen's node bisects the ectoderm in two bilateral neural plates leaving in its wake the floor plate, the notocord and the dorsal endoderm. PMID- 9853822 TI - Using EC and ES cell culture to study early development: recent observations on Indian hedgehog and Bmps. AB - Despite great technological advances in the study of mammalian development in the past two decades, certain problems in early development, such as how the extraembryonic lineages are established, have remained intractable. We suggest that teratocarcinoma (EC) and embryonic stem cells (ES) remain useful in vitro tools for studying some of these problems. We present a continuation of our studies on the role of IHH-based signaling in early development and demonstrate that the IHH N-peptide is expressed in the outer visceral endoderm cells of both the EC and ES-derived embryoid body. We also show that Bmp2 is upregulated and Bmp4 downregulated during the differentiation of F9 EC cells into embryoid bodies, whereas both genes are upregulated when J7 ES cells differentiate into embryoid bodies. We also examine the spatial localization of Ihh, Bmp2, and Bmp4 in day 6.5-7.0 and 7.5-8.0 embryos by in situ hybridization analysis. These data support the EC temporal expression data in that all 3 genes are expressed in visceral endoderm. Bmp4 expression appears to be limited to extraembryonic regions, where mesoderm as well as visceral endoderm are stained. Ihh and Bmp2 are expressed in extraembryonic tissues and the embryo proper. Functional roles for the observed expression patterns are discussed. PMID- 9853823 TI - Embryonic stem cells and transgenic mice in the study of hematopoiesis. AB - Blood formation (hematopoiesis) entails the generation of hematopoietic stem cells (HSCs) within the embryo and subsequent commitment of multipotential progenitors to differentiation along single lineages. These processes are controlled in large part by cell-restricted transcription factors which cooperate with more widely expressed factors to direct lineage-specific gene expression. Candidate hematopoietic transcriptional regulators have been identified by characterizing factors mediating cell-specific gene transcription and by defining genes involved in chromosomal rearrangements in leukemia. The application of transgenic and embryonic stem cell methods have provided insights into their in vivo functions and suggested mechanisms by which lineage selection may be achieved. One of the first, and best, characterized hematopoietic transcription factors is GATA-1. Herein studies of GATA-1 are reviewed to illustrate how manipulations of its locus in the mouse have contributed to current understanding in unique and unexpected ways. PMID- 9853824 TI - Liver disease and compensatory growth: unexpected lessons from genetically altered mice. AB - Over the last decade, several animal models have been established that permit exploration of liver biology and disease. Although these models have been developed using diverse strategies, including transgene targeting, homozygous gene disruption and administration of hepatotoxic chemicals, each approach creates an animal with hepatocyte damage, resulting in an hepatic microenvironment that supports proliferation of healthy hepatocytes. These models have been used to demonstrate: (1) the remarkable ability of adult hepatocytes to clonally proliferate in response to liver growth signals, (2) the effectiveness of transplanted donor hepatocytes in repopulating damaged liver parenchyma, and (3) the feasibility of reconstituting liver with xenogeneic hepatocytes. This paper reviews the development and use of these models, and outlines their potential future application to the study of hepatic stem cells, therapy of liver disease and hepatic toxicology. PMID- 9853825 TI - Thirteen years of manipulating the mouse genome: a personal history. AB - In 1974, Dr. Ralph Brinster published a paper describing the consequences of injecting embryonal carcinoma cells, the predecessors of embryonic stem cells, into mouse blastocysts. Despite their early promise, embryonal carcinoma cells would not efficiently populate the germ line of mice. A decade later Elizabeth Robertson and I described the efficient generation of germline chimaeras from cultured embryonic stem cells and shortly afterwards the genetic manipulation of the mouse germline using ES cells. Our demonstration of the potency of Embryonic Stem cells gave birth to a new era in manipulative mouse genetics, one in which endogenous genes can now be mutated at will using gene targeting of retroviral mutagenesis. This review focuses on the development and testing of concepts and techniques during the thirteen years after we knew germline modification of endogenous genes in the mouse would be possible. This period is one in which more and more sophisticated tools for manipulating the mouse germline were developed and implemented. In this review I have taken the rare opportunity to reveal some of my thought processes, frustrations, successes and failures as we moved through this exciting period of rapid technological change. As I look forward to the next thirteen years, I feel that this will be an equally exciting period for manipulative genetics as we struggle to formulate concepts and design experiments that enable us to understand gene function in an era when the sequence of all genes will be known. PMID- 9853826 TI - Imprinting. AB - Imprinting provides a fascinating mechanism of control of gene expression so that the maternal and paternal alleles of some genes are unequally expressed. Imprinting is most likely established during gametogenesis by a mechanism not completely clear, though DNA methylation probably plays a certain role. Expression of imprinted gene significantly affects mammalian development so that only the maternal or only the paternal diploid genomes cannot support normal development. Since imprinting results in functional hemizygocity, mutation of the expressed allele can have the drastic consequences of a null mutation. For this reason identification of imprinted genes and further understanding of the imprinting mechanism represent an important change for human medical genetics. PMID- 9853827 TI - Genetics of mouse growth. AB - During development, mammalian organisms increase in size until a limit is reached that is mainly determined by the rate and duration of occurrence of cellular divisions increasing total cell number. This process is mostly regulated by an orchestration of the actions of genes participating in pathways that promote or inhibit growth through systemic or local effects. This view of growth control genes and of their effects on the cell cycle has begun emerging from the results of transgenic and gene knockout experiments, which have also re-emphasized the central involvement of some growth factors and hormones in growth signaling, although mechanistic relationships and details about the coordination of growth with patterning, differentiation and morphogenesis continue to remain largely elusive. PMID- 9853828 TI - Immunoglobulin transgenes as targets for somatic hypermutation. AB - This review describes studies on somatic hypermutation of immunoglobulin genes that were started in the mid-80s in collaboration with Ralph Brinster. Almost all of the experiments were carried out using Ig transgenes as targets for the somatic mutation mechanism. Ig transgenes can be very good targets of somatic mutation, despite many different transgene integration sites. Thus, the required cis-acting elements must be present within the approximately 10 kb of the transgene. Only the Ig variable region and its proximate flanks are mutated, not the constant region in unmanipulated sequences. Several Ig gene enhancers are permissive for somatic mutation and they do not have to be associated with the Ig promoter they normally interact with. However, the mutation process does seem to be specific for Ig genes. No mutations were found in several housekeeping genes isolated from cells that had very high levels of somatic hypermutation of their Ig genes. This suggests that the Ig enhancers provide the lg gene specificity. An exception is the Bcl-6 gene, encoding a transcription factor, which was found to be mutated in normal human memory B cells. When the transcriptional promoter that is located upstream of the variable region is duplicated upstream of the constant region, this region is mutated as well. This suggests a transcription coupled model in which a mutator factor associates with the RNA polymerase at the initiation of transcription, travels with the polymerase during elongation, and causes mutations during polymerase pausing. Our recent data with an artificial substrate for somatic mutation suggest that the mutations are increased by increased stability of the secondary structures in the nascent RNA, and the specific nucleotides that are mutated are due to preferences of a mutator factor. PMID- 9853829 TI - Analysis of transcriptional regulatory regions in vivo. AB - Understanding the transcriptional regulation of development and tissue-specific gene expression is a central goal of modern biology. Although the analysis of gene transcription in transfected cultured cells has been essential in establishing many key aspects of this gene control, only analysis in animals can determine developmental timing and cell-specificity of expression within a complex organ and in all the tissues of an animal. The advent of transgenesis made in vivo studies possible. A summary of the in vivo regulatory properties of the pancreas-specific transcriptional enhancer of the rat elastase 1 gene (ELA1) and the role individual elements in this enhancer play in directing high level, cell-specific transcription illustrates the nature, revelations and limitations of transgenic analysis. PMID- 9853830 TI - Angiogenesis and apoptosis are cellular parameters of neoplastic progression in transgenic mouse models of tumorigenesis. AB - The epidemiology and histopathology of human cancers and studies of animal models of tumorigenesis have led to a widely-accepted notion that multiple genetic and epigenetic changes have to accrue for the successful development of a malignant phenotype. Tumor growth and expansion requires an ability not only to proliferate, but also to down-modulate cell death (apoptosis) and activate angiogenesis to produce a tumor neovasculature. This review will describe the interplay between apoptosis and proliferation, as well as the characteristics of the angiogenic phenotype in two transgenic mouse models of multi-step tumorigenesis, namely, pancreatic islet cell carcinomas and squamous cell carcinomas of the skin. PMID- 9853831 TI - The use of transgenic animals in biotechnology. AB - The development of a technique that allows for oocyte and early embryo manipulation is one of the major scientific endeavors in the field of genetic manipulation for animal disease models, basic science in gene regulation and commercial applications. Dr. Ralph Brinster is one of the most prestigious scientists in the development of this science. Through his direction and support, the undertaking of the mechanisms that are involved in the earlier stages of embryology have been productive and enlightening. This paper outlines just some of the experimental successes that evolved from Dr. Brinster's insight and mentorship of one of his pupils. The essay outlines several experimental approaches that have contributed to this field. Specifically, it addresses how the mouse oocyte and the zygote respond to messenger RNA when introduced into the cell, in comparison to comparable non-mammalian species embryos. In addition, this paper discusses some transgenic animal models, both from a basic science point of view and a commercial extension of these techniques. This extension of Dr. Brinster's pioneering work is through technology that allows for the introduction of foreign DNA that can be expressed in targeted organs, such as the mammary gland for production of pharmaceuticals for use in clinical applications. PMID- 9853832 TI - Insertional mutagenesis in transgenic mice generated by the pronuclear microinjection procedure. AB - Insertional mutagenesis in transgenic mice is a powerful method to study structure/function relationships between individual genes and complex developmental traits in the whole organism. Unlike spontaneous or chemical induced mutations, insertional mutations have the advantage that the mutant locus is "tagged" with the transgene and, therefore, readily accessible at the molecular level. Starting with the work on the limb deformity locus, we describe here the characterization of several mouse mutants generated by insertional mutagenesis with the pronuclear microinjection procedure. These transgenic lines have proven to be ideal as models for human disease and for studying the function of novel genes during development. We also describe the unique features of insertional mutations that arise in transgenic mice produced with the pronuclear microinjection procedure and provide recommendations on how to clone and characterize these mutations at the molecular level. Finally, we discuss future prospects for the use of this unique form of germline mutagenesis in the mouse. PMID- 9853833 TI - Transgenic studies of peripheral and central glia. AB - Transgenic manipulation of gene expression in the nervous system has proven immensely useful for the study of glia. This review focuses on studies of Schwann cell and astrocyte biology and pathology. These studies began with promoter mapping for glial-specific genes (P0 and GFAP), and then progressed to oncogene induced transformation and toxin-induced cell ablation of glia. For GFAP, an intermediate filament of astrocytes, we have investigated the effects of alterations in gene dosage, both in terms of deficiency or excess of this structural protein. Finally, the utility of green fluorescent protein as a marker for live astrocytes is described. PMID- 9853834 TI - Comprehensive mammalian genetics: history and future prospects of gene trapping in the mouse. AB - Gene trapping has matured into a tool with tremendous potential for mammalian biology. It both mutates and helps identify genes and can be streamlined so that many thousands of insertions can be characterized. In only a few years most of the genome of the mouse will be tagged and mutated using the latest gene trap designs. By creating such a resource, costly and time consuming alternative methods of mutagenesis and gene identification can be avoided allowing biologists to concentrate on determining gene function in vivo. This will mean a major shift in how the genome will be mined for new drug targets. Notably, gene discovery via gene traps does not suffer from the limitations of other methods as it is not biased by expression level. Mouse strains with specific gene mutations can be easily derived from a gene trap library constructed using embryonic stem cells. These strains will help determine the role of the gene product in mammalian physiology and hence the relevance of the gene product to human disease. PMID- 9853835 TI - Developmental fates of the mouse germ cell line. AB - The specification of mouse germ cell lineage takes place after a population of pluripotential cells is established, and cell communication among the pluripotential cells may be important for this process. Primordial germ cells (PGCs) first appear around the allantois at 7 dpc which are distinct from pluripotential cells in the early embryo because they can not colonize blastocysts. However, a portion of PGCs are transformed into pluripotential cells in the ectopic environment or in culture, suggesting that the developmental fate of PGCs may still be somewhat plastic. PGCs may be destined only for gametes after they enter into the mitotic arrest phase or the meiotic prophase in embryonic gonads, which may be regulated by intrinsic and/or environmental molecules. After fetal germ cells are mitotically arrested, a large number of germ cells undergo programmed cell death. Bcl-2 and its related molecules are involved in the determination of death or survival of fetal germ cells, as well as of spermatogonia in adult testis. The cell death of spermatogonia may be necessary either for eliminating impaired germ cells or for arranging optimal interactions between germ cells and their supporting cells. Although maturating germ cells seem to differentiate only to sperm cells, oocytes that complete the first meiotic division can give rise to pluripotential cells, suggesting that maternal molecules accumulated in oocyte may play a role in the restoration of pluripotency. PMID- 9853836 TI - The germ cell--the mother of all stem cells. AB - The germline, uniquely amongst the lineages of the embryo, carries the genome from generation to generation and is therefore the only lineage which retains true developmental totipotency. Paradoxically, when mouse primordial germ cells (PGCs) are introduced into a host blastocyst, they do not contribute to either the germline or the soma, suggesting that they are restricted in developmental potency. Conversely, in vivo PGCs give rise to embryonal carcinoma (EC) cells, the pluripotent stem cells of teratomas, benign tumors containing derivatives of the three primary germ layers. Similarly, PGCs can be converted in vitro into embryonic germ (EG) cells, pluripotent stem cells capable of giving rise to somatic and germline chimeras. The ability of PGCs to form EC cells in vivo and EG cells in vitro suggests that developmental potency of PGCs is regulateable. The molecular mechanisms controlling PGC growth and differentiation are gradually being elucidated through the characterization of sterile mutants and through the use of in vitro culture systems. Understanding how a PGC can give rise to a pluripotent stem cell could give significant insights into the regulation of developmental totipotency as well as having important implications for male fertility and the etiology of testicular cancer. PMID- 9853838 TI - Male germ cell transplantation: present achievements and future prospects. AB - Germ cells are unique, since their surviving descendants can undergo meiosis and differentiate into gametes, which transmit genetic material from one generation to another. We now know that male germ cells, whether they be primordial germ cells in gonadal ridges, gonocytes, or stem spermatogonia, are transplantable. The donor cells can be transferred by direct microinjection into the seminiferous tubules, rete testis or efferent ducts, depending on the recipient species. Following transplantation, the donor cells undergo spermatogenesis in the host's seminiferous tubules in rats and mice, and have even sired offspring in mice. Interspecific germ cell transfer is possible if the recipient's immune system is defective; nude or SCID mice can even produce rat spermatozoa. However, the major obstacle restricting widespread use of this new technology is its extremely low success rate. This article discusses some ideas for improving the success rate of the transfer technique, and considers several potential applications. PMID- 9853839 TI - Venous thromboembolism and the pill. The WHO technical report on cardiovascular disease and steroid hormone contraception: state-of-the-art. World Health Organization. PMID- 9853840 TI - Is antioxidant therapy a promising strategy to improve human reproduction? Are anti-oxidants useful in the treatment of male infertility? PMID- 9853837 TI - A systematic molecular genetic approach to study mammalian germline development. AB - It is difficult to study gene expression in mammalian embryonic germ cells as PGCs constitute only a minor proportion of the mouse embryo. We have overcome this problem by using a novel combination of established molecular and transgenic approaches. A line of mice has been generated in which the cells of the germ lineage express the beta-galactosidase reporter gene during embryogenesis. Using this line, germ cells have been purified to near homogeneity from embryos at discrete stages during germline development by use of a stain for beta-gal activity and a fluorescence activated cell sorter. Subsequently, cDNA libraries have been constructed from each germ cell population using a modified lone-linker PCR strategy. These combined cDNA libraries represent genes expressed in PGCs during mammalian germline development. To facilitate a molecular genetic approach to studying mammalian germline development, these cDNA libraries will be pooled to form an arrayed, addressed reference embryonic germ cell cDNA library. In parallel with large-scale cDNA sequencing efforts; genes that are differentially expressed in germ cells will be identified by screening the reference library with probes generated by subtractive hybridization. Complementary DNAs identified using this approach will be analyzed by sequencing, database comparison, genomic mapping and in situ hybridization to ascertain the potential functional importance of each gene to germline development. In addition to providing a wealth of novel information regarding patterns of gene expression during mammalian germline development, these results will form the basis for future experiments to determine the function of these genes in this process. PMID- 9853841 TI - Future use of clomiphene in ovarian stimulation. Psychic effects of clomiphene citrate. PMID- 9853842 TI - Embryo transfer and multiple gestation. How many transfers are too many? PMID- 9853843 TI - The effect of ovarian steroids on epithelial ciliary beat frequency in the human Fallopian tube. AB - Using a method that detects variations in light intensity we have studied the effect of ovarian steroids on human Fallopian tube epithelial ciliary beat frequency in vitro. We have found that baseline ciliary beat frequency averages between 5-6 Hz. Cilia from ampullary segments of the Fallopian tube beat significantly faster (5.4 Hz+/-0.2) than those from fimbrial segments (4.8 Hz+/ 0.2). There was no significant difference in baseline ciliary beat frequency at any other anatomical site in the Fallopian tube. Incubation with progesterone (10 micromol/l) suppresses human Fallopian tube epithelial ciliary beat frequency by 40-50%. This inhibition was observed at similar magnitudes in all Fallopian tubes studied irrespective of anatomical site. Progesterone-induced reductions in ciliary beat frequency were concentration dependent and prevented by the progesterone receptor antagonist mifepristone (RU486). Oestradiol alone (10 micromol/l) had no effect on ciliary beat frequency at any anatomical site in the Fallopian tube but did prevent the reduction in ciliary beat frequency seen with progesterone when tissues were incubated with these two steroids together. PMID- 9853844 TI - A prospective randomized clinical trial comparing recombinant follicle stimulating hormone (Puregon) and human menopausal gonadotrophins (Humegon) in non-down-regulated in-vitro fertilization patients. AB - A randomized clinical trial was performed comparing recombinant follicle stimulating hormone (rFSH, Puregon, n = 54) and human menopausal gonadotrophin (HMG, Humegon, n = 35) in infertile women undergoing in-vitro fertilization without the use of a gonadotrophin-releasing hormone (GnRH) agonist. Most patients had a tubal or idiopathic infertility, the latter always longer than 4 years' duration. Patients with sperm abnormalities were excluded. None of the between-group differences in treatment outcome was statistically significant. In the rFSH group, a mean number of 11.2 oocytes was retrieved compared with 8.3 in the HMG group. Ongoing pregnancy rates per started cycle were higher in the rFSH group (22.2%) than in the HMG group (17.1%). Implantation rates were 27.5% in the rFSH group in comparison with 16.7% in the HMG group. In the rFSH group, a mean total dose of 1410 IU during 6.2 days was administered compared with 1365 IU in 6.0 days in the HMG group. Oestradiol concentrations on the day of human chorionic gonadotrophin administration were 3889 pmol/l in the rFSH group and 3145 pmol/l in the HMG group. In 15 subjects (rFSH: n = 9, 16.7%; HMG: n = 6, 17.1%) luteinizing hormone concentrations higher than 10 IU/l were seen during stimulation. In two of them, both from the rFSH group, ongoing pregnancies were achieved. The results indicate that rFSH (Puregon) is at least as efficacious as HMG and that acceptable pregnancy rates can be achieved without the use of a GnRH agonist. PMID- 9853845 TI - The ratio of 2nd to 4th digit length: a predictor of sperm numbers and concentrations of testosterone, luteinizing hormone and oestrogen. AB - The differentiation of the urinogenital system and the appendicular skeleton in vertebrates is under the control of Hox genes. The common control of digit and gonad differentiation raises the possibility that patterns of digit formation may relate to spermatogenesis and hormonal concentrations. This work was concerned with the ratio between the length of the 2nd and 4th digit (2D:4D) in humans. We showed that (i) 2D:4D in right and left hands has a sexually dimorphic pattern; in males mean 2D:4D = 0.98, i.e. the 4th digit tended to be longer than the 2nd and in females mean 2D:4D = 1.00, i.e. the 2nd and 4th digits tended to be of equal length. The dimorphism is present from at least age 2 years and 2D:4D is probably established in utero; (ii) high 2D:4D ratio in right hands was associated with germ cell failure in men (P = 0.04); (iii) sperm number was negatively related to 2D:4D in the right hand (P = 0.004); (iv) in men testosterone concentrations were negatively related to right hand 2D:4D and in women and men LH (right hand), oestrogen (right and left hands) and prolactin (right hand) concentrations were positively correlated with 2D:4D ratio and (v) 2D:4D ratio in right hands remained positively related to luteinizing hormone and oestrogen after controlling for sex, age, height and weight. PMID- 9853846 TI - Critical ovarian hyperstimulation syndrome in a coasted in-vitro fertilization patient. AB - We report an instance of critical ovarian hyperstimulation syndrome in a highly responsive in-vitro fertilization patient despite the preventive measure of a 4 day 'coast' interval during which no gonadotrophins were administered while gonadotrophin-releasing hormone agonist therapy continued until serum oestradiol concentrations fell below 3000 pg/ml. PMID- 9853847 TI - Involvement of ovarian kinin-kallikrein system in the pathophysiology of ovarian hyperstimulation syndrome: studies in a rat model. AB - The purpose of the present study was to investigate a possible participation of the kinin-kallikrein system (KKS) in the pathophysiology of ovarian hyperstimulation syndrome (OHSS). Symptoms of hyperstimulation were produced in immature female rats using equine chorionic gonadotrophin followed by human chorionic gonadotrophin (HCG). At 48 h after the HCG injection, rats were injected s.c. with 100 microg/kg of HOE140, bradykinin-2 receptor antagonist. Capillary permeability was evaluated using peritoneal Evans blue dye (EB) concentrations 30 min after the i.v. injections. The EB concentrations in the hyperstimulated rats were significantly reduced 4 and 6 h after the HOE140 injection, compared with those injected with the vehicle as a control (4.58+/ 0.80 versus 8.22+/-0.87 and 4.32+/-0.74 versus 8.35+/-1.03 microg respectively; P < 0.03), indicating the involvement of kinin in the pathophysiology of OHSS in this model. The administration of 10 IU aprotinin significantly reduced the peritoneal EB concentration when compared with the control (4.13+/-0.53 versus 7.95+/-1.06 microg; P < 0.01), implicating a possible role of kallikrein. Furthermore, pretreatment with RU486 (5 or 10 mg/kg) resulted in a significant reduction of ovarian kinin concentrations 48 h after the HCG injection, compared with the control (1.22+/-0.07 or 1.43+/-0.07 versus 1.94+/-0.10 pg/mg; P < 0.005 and P < 0.05 respectively). Similar results were obtained in the peritoneal EB concentrations. In addition, a significant correlation between the ovarian kinin and peritoneal EB concentrations was observed (P < 0.001, r = 0.539). Thus it was suggested that ovarian KKS plays an intermediary role in the progesterone-induced augmentation of capillary permeability in this experimental model, indicating the involvement of KKS in the pathophysiology of OHSS. PMID- 9853848 TI - Incipient ovarian failure associated with raised levels of follicle stimulating hormone and reduced levels of inhibin A in older sheep. AB - In women there is a gradual rise in the basal level of follicle stimulating hormone (FSH) in the years prior to the menopause (pre-menopause) which is thought to be due to a relative lack of ovarian factors reflecting the number of antral follicles present in the ovaries. Experimental animal models for this phenomenon, particularly in mono-ovulatory species, have been lacking due to most animals' relatively short life span. We have available a group of experimental ewes in which the right ovary was removed and the left ovary was autotransplanted to the neck at least 10 years previously, which have been maintained in good health until an age of 12-13 years. Two experiments were conducted with these animals to determine the endocrine and follicular effects of age: a retrospective experiment in the same Finn-Merino ewes (n = 5) when the animals were 6-7 or 12 13 years of age; and a cohort experiment in old (12-13 years, n = 6) and young (2 years, n = 5) ewes of the same breed. In both retrospective and cohort experiments, the concentrations of FSH were significantly higher (P < 0.05) in older animals during the luteal phase when oestradiol secretion was low. This increase in FSH was associated with a decrease in the concentration of inhibin A (P < 0.05) in older animals in both the follicular and luteal stages of the cycle but the concentrations of oestradiol were similar between ages. Although there were significantly fewer antral follicles (P < 0.05) available for development in older ewes during the early luteal phase of the cycle, the ovulation rate was similar to that observed in younger animals (2.0+/-0 vs 2.0+/-4; P > 0.05) but the interval from luteal regression to the onset of the LH surge was longer (P < 0.05) in older animals. In conclusion, the endocrine changes associated with increasing reproductive age in sheep are therefore similar to those observed in women, suggesting that the sheep could be a useful animal model to study the effect of age on human fertility. PMID- 9853849 TI - A double-blind, randomized, dose-finding study to assess the efficacy of the gonadotrophin-releasing hormone antagonist ganirelix (Org 37462) to prevent premature luteinizing hormone surges in women undergoing ovarian stimulation with recombinant follicle stimulating hormone (Puregon). The ganirelix dose-finding study group. AB - A multicentre, double-blind, randomized dose-finding study of Org 37462 (ganirelix) was conducted in 333 women undergoing ovarian stimulation with recombinant follicle stimulating hormone (rFSH; Puregon) to establish the minimal effective dose preventing premature luteinizing hormone (LH) surges during ovarian stimulation. For ovarian stimulation, rFSH was given in a fixed daily dose of 150 IU for 5 days from days 2 to 6 of the menstrual cycle. From cycle day 7 onward, up to and including the day of human chorionic gonadotrophin (HCG), Org 37462 (dosages 0.0625, 0.125, 0.25, 0.5, 1.0 and 2.0 mg/0.5 ml) was administered once daily by s.c. injection, and the rFSH dose was adjusted depending on ovarian response. The lowest (0.0625 mg) and highest (2.0 mg) dose groups were terminated prematurely on the advice of an external independent advisory committee. Serum Org 37462 concentrations increased in a linear dose-proportional manner, whereas serum LH and increases of oestradiol fell with increasing Org 37462 dose. During Org 37462 treatment, serum LH concentrations > or =10 IU/l were observed in the lowest dose groups with incidences of 16% (0.0625 mg), 9% (0.125 mg) and 1.4 % (0.25 mg). On the day of HCG, the number of follicles > or =11, > or =15 and > or =17 mm were similar in the six dose groups, whereas serum oestradiol concentrations were highest in the 0.0625 mg group (1475 pg/ml) and lowest in the 2 mg group (430 pg/ml). The median daily dose of rFSH was between 150 and 183 IU and the overall median duration of Org 37462 treatment was approximately 5 days in the six treatment groups. Overall, Org 37462 treatment appeared to be safe and well tolerated. The mean number of recovered oocytes and good-quality embryos was similar in all dose groups and ranged from 8.6 to 10.0 and 2.5 to 3.8, respectively. The mean number of replaced embryos in the different dose groups ranged from 2.3 to 2.7. The implantation rate was highest in the 0.25 mg group (21.9%) and lowest in the 2 mg group (1.5%). The early miscarriage rates (first 6 weeks after embryo transfer) were 11.9 and 13% in the 1 and 2 mg group respectively, whereas in the other dose groups this incidence was zero (0.0625%) up to a maximum of 3.7% (0.5 mg group). The vital pregnancy rate (with heart activity) at 5-6 weeks after embryo transfer was highest in the 0.25 mg group, i.e. 36.8 % per attempt and 40.3 % per transfer, and resulted in an ongoing pregnancy rate 12-16 weeks after embryo transfer of 33.8% per attempt and 37.1% per transfer. In conclusion, a daily dose of 0.25 mg Org 37462 prevented LH surges during ovarian stimulation and resulted in a good clinical outcome. PMID- 9853850 TI - Male infertility: analysis of the markers and genes on the human Y chromosome. AB - The long arm of the human Y chromosome is required for male fertility. Deletions in three different regions can cause severe spermatogenic defects ranging from non-obstructive azoospermia to oligozoospermia. Use of intracytoplasmic sperm injection (ICSI) may allow Y chromosome defects to be passed from father to son. Thus, numerous reports have stressed the need to offer genetic testing to infertile men who select ICSI and a number of reproductive clinics have begun to do so. The primary objectives of this review were: firstly, to discuss the characteristics of the published set of polymerase chain reaction markers and how these characteristics affect interpretation of Y chromosome deletion analysis and secondly, to summarize the recent literature pertaining to the genes on the Y chromosome. PMID- 9853851 TI - A familial case of X chromosome deletion ascertained by cytogenetic screening of women with premature ovarian failure. AB - The association between X chromosome deletions and premature ovarian failure is well established. Previous anecdotal reports however, have not documented the prevalence of X deletions in women with premature ovarian failure. We therefore performed cytogenetic analyses on 79 women with primary or secondary amenorrhoea to assess the utility of screening for a genetic marker for familial premature ovarian failure. A normal karyotype was found in 77 women. One woman with primary amenorrhoea had an XY karyotype and a woman with secondary amenorrhoea had a deletion at Xq 26.1. This second case had a family history of premature ovarian failure, and her mother who underwent premature ovarian failure at 28 years shared this deletion. The early diagnosis of familial X deletions causing premature ovarian failure allowed for the prediction of impending menopause and the implementation of manoeuvres to advance conception. Although cytogenetic aberrations are rare in secondary amenorrhoea, the ability to predict premature ovarian failure can be vital. PMID- 9853852 TI - Intramesosalpingeal injection of oxytocin in conservative laparoscopic treatment for tubal pregnancy: preliminary results. AB - The objective of this study was to evaluate the efficacy of the local use of oxytocin as adjuvant treatment in conservative surgery for tubal pregnancy. The patients were 25 women with laparoscopic diagnosis of tubal pregnancy who were candidates for salpingotomy. Before salpingotomy, each patient was randomly allocated to intramesosalpingeal injection of 20 IU oxytocin diluted to 20 ml with saline solution or 20 ml saline solution. The surgeon then proceeded with salpingotomy and removal of the pregnancy according to the usual technique. The main outcome measures were bleeding during salpingotomy, ease of removal of the pregnancy, bleeding at the site of the pregnancy, and need for recourse to salpingectomy. Twelve women were randomized to the oxytocin group and 13 to the control group. Examination of the surgeons' assessments of the difficulties encountered at the different stages of surgery revealed statistically significant differences between the oxytocin group and controls in each variable. In particular, the amount of endosalpingeal bleeding after removal of the pregnancy was less in the oxytocin group. In one control patient, persistent bleeding due to incomplete trophoblast removal forced the surgeon to perform salpingectomy. Our results indicate that intramesosalpingeal injection of oxytocin facilitates the performance of conservative laparoscopic treatment for tubal pregnancy. PMID- 9853853 TI - A 3 year retrospective review of intrauterine insemination, using cryopreserved donor spermatozoa and cycle monitoring by urinary or serum luteinizing hormone measurements. AB - Insemination with donor spermatozoa is an integral part of infertility treatment. For the last 3 years in our unit, intrauterine insemination with donor spermatozoa (IUID) has been used in preference to vaginal insemination. In this retrospective study, patients were offered an initial course of five single intrauterine inseminations with cryopreserved donor spermatozoa and treatment was then reviewed. A total of 389 patients received 1465 inseminations. In all, 1119 cycles were monitored using luteinizing hormone serum analyses and 346 cycles using the urine home test kits. The clinical pregnancy rate per insemination for the cycles monitored by the serum assay was 18.0% (202/1119) compared with the urine cycles (13.7%, 46/346) (P <05). The pregnancy loss rate was not significantly different (14.4%, 29/202 and 21.7%, 10/46) (serum and urine cycles respectively). The viable clinical pregnancy rate was significantly higher (P <03) for the serum cycles than for the cycles using the urinary monitoring (15.5%, 173/1119 and 10.4%, 36/346 respectively). The cycles monitored by serum assay had a significantly higher cumulative viable clinical pregnancy rate (P <0001) of 70.2% after nine inseminations compared with the urine monitored cycles of 54.8%. The majority of patients opted for the serum cycles, with a minority self-selecting the urine cycles mainly for travelling convenience. The explanation for the significant differences between the viable clinical pregnancy rates per insemination and the cumulative viable clinical pregnancy rates may be due to the sensitivity of the urine home test kit or the patients' interpretation of the result. PMID- 9853854 TI - Hysterosalpingo contrast sonography (HyCoSy) with SH U 454 (Echovist) for the assessment of tubal patency. AB - A total of 88 Fallopian tubes from 44 patients was examined with hysterosalpingo contrast sonography (HyCoSy), hysterosalpingogram (HSG), and laparoscopic chromopertubation (LC) in order to assess their relative accuracy for measuring tubal patency. HyCoSy was done by transvaginal ultrasound and the contrast was SH U 454 (Echovist). The flow of multiple fractions of the contrast medium through each Fallopian tube was observed in real time in appropriate imaging planes by means of a transvaginal probe. Compared with laparoscopic results, we found a sensitivity of 85.2%, a specificity of 85.2%, a positive predictive value (PPV) of 71.9%, a negative predictive value (NPV) of 92.9% and concordance (HyCoSy/LC) of 85.2%, while the corresponding values for HSG were sensitivity = 85.2%, specificity = 83.6%, PPV = 69.7%, NPV = 92.7% and concordance (HSG/LC) of 84.1%. Compared with HSG results, HyCoSy obtained a co-positivity of 66.7%, a co negativity of 81.8% and a concordance of 76.1%. In conclusion, HyCoSy with SH U 454 proved to be a reliable and safe modality for evaluating tubal patency; it is suitable as an outpatient diagnostic procedure to be used before more invasive procedures. PMID- 9853855 TI - Questioning the efficacy of Fallopian tube sperm perfusion. AB - The aim of this work was to compare the efficiency of standard intrauterine insemination (IUI) and Fallopian tube sperm perfusion (FSP) in the treatment of infertility. Ninety-six consecutive patients with infertility in 100 cycles were included in the study. Those randomized to standard IUI included 48 patients in 50 cycles [25 clomiphene citrate only and 25 clomiphene citrate/human menopausal gonadotrophin (HMG) cycles] (group I). Patients subjected to FSP included 48 patients in 50 cycles (18 clomiphene citrate only and 32 clomiphene citrate/HMG cycles) (group II). The overall pregnancy rate per cycle (16% versus 18%) was not significantly different in the two groups. The pregnancy rates were also similar in the two groups when compared for the cause of infertility: ovulatory disorder 16.7% versus 16%, tubal impairment 10% versus 9.1%, cervical hostility (no pregnancy occurred in this group) and unexplained infertility 21.4 % versus 25 %. The overall pregnancy rate (for the two groups) appeared higher when clomiphene citrate/HMG was used for ovulation induction (21.1%) than when clomiphene citrate only was used (11.6%). PMID- 9853856 TI - A 2-year comparative study of endometrial histology and cervical cytology of contraceptive implant users in Birmingham, UK. AB - The objectives of this study were to evaluate the endometrial histology and cervical cytology of users of two contraceptive implants releasing etonogestrel/3 keto-desogestrel (Implanon) and levonorgestrel (Norplant) in West Midlands (UK) users. A 2-year prospective randomized design was used to study 60 implant users. Endometrial histology and cervical cytology were compared before insertion and after 12 and 24 months. At the end of 12 months, the majority of samples were inactive/weakly proliferative in both groups. At the end of 24 months, this remained unchanged in the Implanon group whereas the pattern was more diverse in the Norplant group. Endometrial thickness was significantly reduced in both groups during treatment. Cervical cytology remained unchanged. It is concluded that, after 2 years, there was no evidence of an increasing risk of endometrial hyperplasia, endometrial carcinoma, cervical intra-epithelial neoplasia or cervical carcinoma in either of the two groups of implant users. PMID- 9853857 TI - Correlation of testicular pathology and sperm extraction in azoospermic men with ejaculated spermatids detected by immunofluorescent localization. AB - Limiting testicular biopsy for intracytoplasmic sperm injection (ICSI) to those with a high chance of having testicular spermatozoa has not been possible because of the poor predictive value of current clinical and laboratory methods. In order to predict testicular pathology and sperm extraction, we characterised the semen of 28 men with azoospermia due to gonadal failure in terms of the presence of spermatids using an immunological method. The results were compared with the assessment of testicular biopsies by histology and the extraction of spermatozoa into culture medium. Washed cellular elements in the ejaculate were smeared on microscope slides and fixed in 100% methanol, before incubation with acrosome specific monoclonal antibody (18.6), fluorescein isothiocyanate-labelled anti mouse goat IgG, and examination by epifluorescent microscopy. Semen from men with oligozoospermia and obstructive azoospermia served as positive and negative controls, respectively. Twelve patients who had positive immunofluorescence (one or more spermatids present) had spermatozoa retrieved from their testes (five hypospermatogenesis, seven focal spermatogenesis), and 16 patients with negative immunofluorescence (spermatids absent) had apparent Sertoli cell-only syndrome (12) or maturation arrest histological pattern (four). However, four patients with apparent Sertoli cell-only syndrome had testicular spermatozoa present after extraction from the biopsy. Plasma follicle stimulating hormone concentration and testicular volume did not predict retrieval of seminal spermatids or testicular spermatozoa. We conclude that the immunofluorescent localization of one or more spermatids in the ejaculate can be used to predict the likelihood of obtaining testicular spermatozoa for ICSI. However, in some patients with Sertoli cell-only syndrome, spermatozoa could still be recovered in the absence of apparent seminal spermatids. PMID- 9853858 TI - Correlation of testicular sperm extraction with morphological, biophysical and endocrine profiles in men with azoospermia due to primary gonadal failure. AB - To identify the predictive factors for testicular sperm extraction (TESE) and to understand the pathology associated with TESE, we carried out a prospective study in 40 consecutive men with azoospermia due to primary gonadal failure. The main outcome measure was the retrieval of at least one testicular spermatozoon. Endocrine and biophysical profiles, testicular histology, Johnsen score and testicular spermatids were used as predictors of sperm extraction. Spermatogenesis was quantified with the Johnsen score. A variable pattern of spermatogenesis was common, being present in 20 (50%) patients. Visualisation of testicular spermatids on testicular histology showed a strong association with TESE (P < 0.0001). Statistically significant differences were detected in plasma follicle stimulating hormone (FSH) and testicular volume between patients who had hypospermatogenesis and Sertoli cell-only or maturation arrest. There were no significant differences in Johnsen score, biophysical and endocrine profiles between the groups with successful and failed TESE. However, a statistically significant trend occurred with changes in histological pattern [chi2 for trend, P = 0.001; Pearson's coefficient (r) = 0.6], Johnsen score (P = 0.022; r = 0.5), testicular volume (P = 0.01; r = 0.5) and plasma FSH concentrations (P = 0.044; r = 0.4), albeit to a limited degree. Difference in the interpretation of histological patterns with different assessors was observed. The type of occupation or risk factors for azoospermia showed no association with testicular pathology or TESE. Variable histological patterns in different tubules in the same individual may explain the poor correlation of TESE with endocrine and biophysical profiles, Johnsen score and histological pattern. Differences in the amount of tissue used for TESE and histopathology, and misinterpretation of testicular histology rather than failure to quantify spermatogenesis may explain the poor correlation between histological patterns and TESE. Testicular spermatids predicted TESE. However, considerable overlap in values means that no single variable can provide a perfect discrimination between the groups with successful and failed TESE. PMID- 9853859 TI - A prospective study of multiple needle biopsies versus a single open biopsy for testicular sperm extraction in men with non-obstructive azoospermia. AB - Little is known about the efficacy and the factors affecting the outcome of fine needle aspiration biopsy of the testis for sperm retrieval in azoospermic men with defective spermatogenesis. A prospective study was designed to compare the efficacy of needle and open (window) testicular biopsies for testicular epididymal sperm extraction (TESE) in 35 consecutive men with azoospermia due to defective spermatogenesis undergoing testicular biopsy for intracytoplasmic injection of oocytes. Each of the consecutive 35 patients underwent TESE using a 19 gauge butterfly needle followed by a window (1-1.5 cm-sized incision) testicular biopsy in the same procedure. The extraction of spermatozoa into culture medium was compared with the assessment of testicular biopsies by histology, the mode of biopsy (needle or open biopsy) and the amount of tissue retrieved by either method. Testicular spermatozoa were retrieved in 22 (63%) who had an open testicular biopsy compared with five (14%) patients who had multiple needle biopsies, respectively; the difference was statistically significant. Open testicular biopsy retrieves more testicular tissue than needle biopsy. Needle testicular biopsy retrieved testicular spermatozoa in 50% of those with hypospermatogenesis, 10% with focal spermatogenesis and in no patients with maturation arrest or Sertoli cell-only pattern. In contrast, sperm retrieval was successful in 100%, 90% and 66% of those with respective histologies using open testicular biopsy. Other than bruising, for which they required no analgesia, none of the patients suffered any obvious complications associated with traditional testicular biopsy. We conclude that open testicular biopsy is more effective than needle biopsy for the retrieval of testicular spermatozoa in azoospermic men with defective spermatogenesis. The difference observed may be related to the amount of testicular tissue retrieved and to the influence of testicular histology. PMID- 9853860 TI - Multiple testicular sampling in non-obstructive azoospermia--is it necessary? AB - Spermatogenesis may be focal in non-obstructive azoospermia. The present study was conducted to determine whether the performance of multiple, rather than a single testicular sample contributes to obtaining spermatozoa in amounts sufficient for fertilization and cryopreservation in non-obstructive, azoospermic patients. Furthermore, the aim was to clarify the significance of location for retrieval from the testis in such cases. Three biopsies were taken from identical locations in 55 testes of 29 men with non-obstructive azoospermia: (i) the rete testis region, ii) the midline, and (iii) the proximal region of the testis. When sperm cells were detected, they were used for intracytoplasmic sperm injection (ICSI), and the remainder were then cryopreserved in as many aliquots as possible (adjusted for ICSI procedure). Spermatozoa were found in 28 testes (50.9%) of 18 men (62.1%). In the testes from which spermatozoa were obtained, they were present in three, two or one locations in 15 (53.6%), five (17.9%) and eight (28.6%) cases respectively. The possibility of finding spermatozoa was not influenced by the location in the testis. Multiple testicular sperm extraction is recommended in cases of non-obstructive azoospermia, since it may enhance diagnostic accuracy of absolute testicular failure and increase the number of sperm cells retrieved. PMID- 9853861 TI - Controlled comparison of percutaneous and microsurgical sperm retrieval in men with obstructive azoospermia. AB - A controlled comparison of the efficacy and reliability of sperm retrieval by testicular fine needle aspiration (TFNA), percutaneous testicular needle biopsy (PercBiopsy) and microsurgical epididymal sperm aspiration (MESA) was performed in nine patients with obstructive azoospermia. During a planned MESA procedure, sperm retrieval was attempted on the same testis with TFNA and PercBiopsy. Spermatozoa were obtained from all patients using MESA and PercBiopsy. Spermatozoa were retrieved using TFNA from 6/9 (67%) men. The mean number of epididymal spermatozoa retrieved with MESA (15 x 106) was significantly higher (P = 0.003) than that retrieved percutaneously from the testis. The mean number of spermatozoa obtained by PercBiopsy was 0.116 x 10(6) while TFNA recovered 0.014 x 106 spermatozoa (P = 0.025). MESA is the optimal choice to retrieve the greatest number of spermatozoa with highest motility for assisted reproduction and subsequent cryopreservation. However, percutaneous testicular retrieval does not require microsurgical expertise and is less invasive. Our results suggest that the optimal percutaneous procedure for sperm retrieval from the testis involves percutaneous testicular needle biopsy with an automatic biopsy gun. PMID- 9853862 TI - Doppler ultrasound of the testis in azoospermic subjects as a parameter of testicular function. AB - Azoospermia frequently represents the end-point of different pathological conditions that cause important quantitative and qualitative alterations of both spermatogenesis and testicular structure, including intratesticular blood vessels. In this study we performed colour Doppler ultrasound of the testis in 12 azoospermic subjects affected by primary testicular pathology (four bilateral post-orchitis, four postradiotheraphy for cancer, four post-traumatic) aged 28.2+/-3.3 (mean+/-SD) years, in six subjects affected by obstructive azoospermia aged 29.7+/-2.4 years and in 20 age-matched fertile subjects (aged 28.6+/-2.5 years). The analysis of intratesticular vessels per organ was quantified using a semiquantitative score: category 0, no vessels visible; category 1, between one and three intratesticular vessels visible; and category 2, more than three vessels visible. In obstructive azoospermic patients and in fertile subjects there were always more than three intratesticular vessels. No intratesticular vessels were detected in eight testes (33.3%) and fewer than three vessels in 16 testes (66.6%) in subjects affected by primary testicular pathology. In azoospermic subjects the testicular structure of the testis was evaluated by diagnostic fine needle aspiration cytology (FNAC) performed in the middle portion of the testis. In non-obstructive azoospermic patients this procedure showed the presence of only Sertoli cells in all cases. When detectable vessels were present, a new aspiration was performed in these areas. In 12 out of 16 cases, spermatogenetic cells including mature spermatozoa, were found when the FNAC was performed in testicular regions showing the presence of blood vessels. These results indicate that colour Doppler sonography of the testis may be useful in the differential diagnosis of azoospermia and suggest the evaluation of the intratesticular blood vessel distribution before performing any method to retrieve intratesticular spermatozoa for intracytoplasmic sperm injection. PMID- 9853863 TI - Cytogenetic analysis of unfertilized oocytes following intracytoplasmic sperm injection using spermatozoa from a globozoospermic man. AB - A man with globozoospermia was treated in our in-vitro fertilization intracytoplasmic sperm injection (ICSI) programme. In the treatment cycle, 24 oocytes were collected from his wife. All the oocytes were at metaphase II stage. The semen sample produced on the day had a normal sperm count, good motility, but with 100% globozoospermia. All oocytes were injected with randomly selected spermatozoa and of these, two oocytes showed two pronuclei and another contained a single pronucleus. The remainder were unfertilized. The normally fertilized oocytes (two pronuclear) cleaved to the four-cell stage and were transferred to the patient. At 48 h after ICSI, the 21 unfertilized oocytes were processed for cytogenetic analysis. All oocytes contained a haploid chromosome set. The only abnormality seen was a chromosome fragment in one metaphase. Eighteen oocytes contained decondensed sperm nuclei and of these, 14 nuclei were beginning to show signs of premature chromatin condensation (PCC) and the other four showed strong signs of PCC. Thus it appears that in some forms of globozoospermia, arrest of nuclear decondensation and/or PCC are another cause of fertilization failure. The most likely cause for this is the absence or down-regulation of spermatozoa associated activating factor in round-headed spermatozoa. PMID- 9853864 TI - Round spermatids from hybrid sterile mice can initiate normal embryo development. AB - Fertilization and embryo development following intracytoplasmic injection of round spermatids collected from hybrid sterile mice (BALB/c) were investigated. The rates of oocyte activation, blastocyst formation and development into live offspring were compared with those after intracytoplasmic injection of round spermatids from fertile mice (B6D2F1). The injection of oocytes with round spermatids from hybrid sterile or sterile males resulted in similar rates of normal fertilization and embryo development. The rates of development to term of 2-cell embryos were also similar, regardless of the origin of the round spermatids injected. This finding suggested that round spermatids from hybrid sterile mice have the ability to fertilize normally and to allow normal embryo development. PMID- 9853865 TI - A first prospective study of the individual straight line velocity of the spermatozoon and its influences on the fertilization rate after intracytoplasmic sperm injection. AB - The straight line velocity of 500 individual spermatozoa was measured immediately before their direct injection into the cytoplasm of 500 metaphase II oocytes. The straight line velocity (VSL) did not have a normal distribution (P < 0.01) and ranged between 0 and 35 microm/s. The fertilization rate (84%) was significantly (P < 0.008) higher in the quartile of the sperm population with the highest VSL compared to the fertilization rate (68%) in the quartile with the lowest VSL. Embryo cleavage and embryo quality were not different in the quartiles of the sperm population used for injection. PMID- 9853866 TI - Influence of oxygen tension on function of isolated spermatozoa from ejaculates of oligozoospermic patients and normozoospermic fertile donors. AB - Oxygen radical generation is known to be detrimental to sperm function. An example of a reactive oxygen species-associated male pathology is oligozoospermia in which fertilization and pregnancy rates are low in in-vitro fertilization (IVF) programmes. As the extent of the modifications induced by reactive oxygen species (ROS) depends on several factors, notably from oxygen tension in the incubation medium, the aim of this study was to examine the influence of a low (5%) rather than atmospheric (20%) oxygen tension in the incubator gas phase on the function of Percoll-selected spermatozoa from ejaculates of oligozoospermic patients and normozoospermic fertile donors. After incubation for several hours in a gas phase of either 5% CO2/90% N2/5% O2 or 5% CO2/95% air (20% O2), none of the parameters investigated, e.g. movement characteristics, potential of spermatozoa to acquire hyperactivated motility, to undergo the acrosome reaction when challenged with a calcium ionophore and to fuse with zona-free hamster oocytes, was significantly different between the two oxygen tensions in fertile donors. In contrast, among oligozoospermic patients, the motility parameters, the percentage of hyperactivated motility and of induced-acrosome reaction were significantly improved under a gas phase of 5% O2 compared with those observed under an atmosphere of 20% O2 (P < 0.05). Exposure to 5% rather than 20% oxygen tension also induced a significant increase in the percentage of penetration of zona-free hamster eggs after capacitation for 17 h, but no difference was found in the mean number of bound spermatozoa per oocyte. After incubation for 24 h, a significantly higher survival rate was observed under 5% compared with 20% oxygen tension. These results show that the use of a low oxygen tension rather than air might improve spermatozoan competence of oligozoospermic patients during IVF programmes. PMID- 9853867 TI - Induction of a differentiated ciliated cell phenotype in primary cultures of Fallopian tube epithelium. AB - Human Fallopian tubal epithelial cells in culture lose morphological features associated with the epithelium in situ and the extent to which they retain their in-vivo phenotype or function is unknown. In order to address this question, immunocytochemical markers were identified which distinguish secretory (HMFG2+, LhS28-) from ciliated (HMFG2-, LhS28+) epithelial cells in tissue sections of Fallopian tube. These markers were used to analyse the phenotype of tubal cells in vitro. Primary cultures of human tubal epithelial cells were seeded onto glass and grown to confluence before addition of oestradiol-17beta. In the absence of hormone, tubal epithelial cells expressed cytokeratins and nuclear receptors for oestrogen and progesterone and adopted a homogeneous (HMFG2+, LhS28-) secretory cell phenotype. Following the addition of oestradiol-17beta, a proportion of cells became positive for LhS28. The induction of a ciliated epithelial cell phenotype was confirmed by scanning electron microscopy, where on permeable collagen membranes, approximately one-third of tubal epithelial cells became ciliated in the presence of oestradiol-17beta. We suggest that in vitro, tubal epithelial cells adopt an immature secretory-like phenotype and that oestrogen can induce differentiation to a ciliated epithelial cell phenotype. PMID- 9853868 TI - Characterization of ciliary activity in distal Fallopian tube biopsies of women with obstructive tubal infertility. AB - Biopsy specimens were obtained from the distal end of the Fallopian tubes of 62 women with tubal infertility and examined by light and electron microscopy. Ciliary beat frequency (CBF) measurements were obtained using laser light spectroscopy. Neither demographic nor behavioural characteristics nor serological evidence of past chlamydial infection were associated with CBF measurements. In contrast, CBF were significantly lower (P < 0.05) in tissues with oedema compared to tissues without oedema (6.7 versus 12.9) and in tissues with erythema compared to tissues without erythema (9.2 versus 13.7). Furthermore, CBF measurements did vary by chlamydial serotype pattern, with lower values observed among the tissues of women with antibodies to serotype C or E (without D) as compared to the tissues of women with other serotypes (P < 0.04). However, these data must be interpreted with caution as the numbers of subjects with chlamydial antibodies to serotype C (n = 3) or E without D (n = 5) were few in number and serotyping of IgG antibodies in blood is not as accurate as it is in bacterial isolates. Confirmation of the suggested association between chlamydial serotype and risk of adverse sequelae could indicate potential new avenues for vaccine research. PMID- 9853869 TI - Febrile morbidity in severe and critical ovarian hyperstimulation syndrome: a multicentre study. AB - The objective of this study was to define the incidence of febrile morbidity and its causes in severe and critical ovarian hyperstimulation syndrome (OHSS). For this purpose, we reviewed the medical records of all OHSS patients hospitalized in 16 out of 19 tertiary medical centres in Israel between January 1987 and December 1996. Febrile morbidity was defined as at least one episode of temperature rise above 38 degrees C lasting > or =24 h. A total of 2902 patients (3305 hospitalizations) with OHSS was identified, of whom 196 had severe, and 13 critical, OHSS. Among the 209 patients investigated the incidence of febrile morbidity was 82.3%, of which 20.5% was attributed to urinary tract infection, 3.8% to pneumonia, 3.3% to upper respiratory tract infection, 2.0% to intravenous line phlebitis, 1.0% to cellulitis at an abdominal puncture site, 1.0% to postoperative wound infections and 0.5 % to gluteal abscess at the site of progesterone injection. Non-typical organisms were frequently isolated, such as Pseudomonas, Proteus, Klebsiella and Enterobacter species. No infectious aetiology was found in 105 patients (50.2%). Hypoglobulinaemia was recorded in most patients, while ascitic and pleural fluids aspirated from these patients contained high globulin concentrations. We conclude that infection-related febrile morbidity in severe and critical OHSS is high, and may be attributed to some degree of immunodeficiency associated with loss of plasma globulins to the third space. However, non-infection-related febrile morbidity is even higher and may be attributed to endogenous pyrogenic mechanisms. PMID- 9853870 TI - Pretreatment with follicle stimulating hormone promotes the numbers of human oocytes reaching metaphase II by in-vitro maturation. AB - The objective of this study was to investigate the effect of follicle stimulating hormone (FSH) priming on the in-vitro maturation (IVM) of human oocytes from healthy ovaries using a chemically defined culture system. Seventeen patients donating oocytes for research received a truncated course of 600 IU FSH over 5 days and a further control group of nine patients received no FSH treatment. Mid follicular phase cumulus-enclosed oocytes (n = 160) were aspirated from follicles < or =4 mm diameter under transvaginal ultrasound guidance and were cultured for 48 h in microdrops of medium containing 10 mIU/ml FSH and 100 mIU/ ml human chorionic gonadotrophin (HCG). The results demonstrated that human oocytes will efficiently undergo IVM under serum-free conditions. After mild FSH stimulation, a greater number of cumulus-enclosed oocytes was collected, and following culture, a lower rate of degeneration was observed. Significantly more oocytes completed nuclear maturation to metaphase II following FSH stimulation (71.1 versus 43.5%). In conclusion, a truncated course of FSH stimulation in vivo improved the oocyte maturation rate in vitro, giving a mean of 4.8+/-0.7 metaphase II oocytes per patient compared with only 2.1+/-0.7 from control patients, thus yielding more mature oocytes for future IVF treatment. PMID- 9853871 TI - Evaluation of three substitutes for Percoll in sperm isolation by density gradient centrifugation. AB - Silane-coated silica particle solutions (ISolate(TM) and PureSperm)TM)) and iodixanol (OptiPrep(TM)) were compared to polyvinylpyrrolidone (PVP)-coated silica particles (Percoll(TM)) in their efficacy to recover spermatozoa by gradient centrifugation for use in assisted reproductive procedures. Efficacy was assessed in terms of percentages of sperm recovery, sperm vitality and motility, normal sperm morphology and normal sperm chromatin condensation. No significant difference was found in the recovery of spermatozoa for men with both normal sperm counts and oligozoospermia, between PVP-coated and silane-coated particle solutions. Iodixanol had significantly lower sperm recovery compared to the other products. Sperm vitality, progressive motility, normal morphology and normal chromatin condensation did not differ significantly between any of the sperm isolation products. PMID- 9853872 TI - Routine addition of human insulin-like growth factor-I ligand could benefit clinical in-vitro fertilization culture. AB - Animal studies suggest that the insulin-like growth factors play an important role in preimplantation embryo development. Human preimplantation embryos express mRNA for insulin-like growth factor-I receptor (IGF-IR) but the ligand, insulin like growth factor-I (IGF-I), is not expressed by the embryo until after implantation. We tested the hypothesis that IGF-I produced by the female reproductive tract may bind to these receptors, augmenting embryo survival, growth and development. Transcripts of mRNA for IGF1 were detected using reverse transcription-polymerase chain reaction in midcycle human Fallopian tube. Immunohistochemistry localized immunoreactive IGF-I to the cytoplasm of all the major structures of the Fallopian tube, with the most intense staining seen in the tubal epithelial lining. Maternally produced IGF-I was present in the fluid found in the human tube and uterus at concentrations of 8.0 and 10.9 nM respectively. Supplementation of culture medium with IGF-I increased the proportion of embryos developing to the blastocyst stage from 35% in controls to 60% in the presence of IGF-I. In addition the total number of cells in day 6 blastocysts was increased by 19% (64.44 versus 54.08% in control, not significant), due entirely to a statistically significant 59% increase (25 versus 15.75%, P = 0.020) in the number of cells in the inner cell mass. The effect of IGF-I was mediated through the IGF-I receptor. Immunocytochemistry using an alphaIR3 antibody confirmed the presence of IGF-I receptor in human blastocysts and the same antibody completely inhibited the stimulation of blastocyst formation by IGF-I. These data suggest that human preimplantation development is enhanced by maternal IGF-I. Mimicking this in-vivo paracrine relationship may improve clinical in-vitro embryo culture and IVF pregnancy rates. PMID- 9853873 TI - Mosaicism in the inner cell mass of human blastocysts. AB - Although mosaicism was shown to be a normal feature in cleaving embryos, its consequences for the late preimplantation stages are unknown. We performed blastocyst immunosurgery, followed by fluorescent in-situ hybridization (FISH), to determine the number of cells and degree of mosaicism in the inner cell mass (ICM) of human blastocysts. Of 47 ICM samples analysed, 20 had aneuploid cells, and two also had a few tetraploid cells. The average degree of aneuploidy in the ICM was similar to the overall blastocyst mosaicism, suggesting that there is probably no selection for euploid ICM. The lower degree of blastocyst mosaicism, compared with the cleavage-stage embryos, may be due to a mechanism of selection against the embryos with high frequency of mosaicism, leading to elimination of these embryos prior to blastocyst formation. PMID- 9853874 TI - Clinical application of human egg cryopreservation. AB - Clinical egg cryopreservation has been applied during a 4-year period with some limited success. Mostly mature and a few immature eggs were frozen slowly and thawed rapidly in 1,2-propanediol and sucrose, and subsequently inseminated by intracytoplasmic sperm injection (ICSI). Three studies were performed in which: (i) it was established that 55% of aged unfertilized mature eggs survive freezing; (ii) in 22 cycles of thawed donated eggs cryosurvival was 24% with 15 cycles reaching transfer, and five pregnancies were initiated, one of which went to term at 39 weeks with fraternal twin boys, and one remains ongoing at 37 weeks; and (iii) in five cycles, where in-vitro fertilization patients had some of their own eggs frozen/ thawed, cryosurvival of mature eggs was poor at only 2.2%, although 44% sibling germinal vesicle (GV) stage eggs survived. A normal female infant delivered at 40 weeks arose from transfer of two embryos where GV eggs underwent in-vitro maturation post-thaw and were fertilized by ICSI. Pregnancies reported here and by others indicate a burgeoning awareness of the potential benefits of egg cryopreservation, prompting cautious optimism for the future of this technology. PMID- 9853875 TI - Effect of timing of oocyte denudation and micro-injection on survival, fertilization and embryo quality after intracytoplasmic sperm injection. AB - In human in-vitro fertilization (IVF), the oocytes are surrounded by cumulus and corona cells at the time of insemination so that their maturity cannot easily be evaluated. The best IVF results are obtained if the oocytes are inseminated 2-6 h after retrieval. In the intracytoplasmic sperm injection (ICSI) procedure, the oocytes are denuded by enzymatic and mechanical treatment in order to be able to perform the injection. As a consequence, the nuclear maturity of the oocytes can be evaluated and only those that have extruded the first polar body are injected. However, metaphase-II oocytes that have not yet reached cytoplasmic maturity cannot be recognized. The purpose of this study was to investigate the effect of different timing of cumulus-corona cell removal and injection on the outcome of ICSI. For this we allowed the oocytes to complete in-vitro cytoplasmic maturation in two different culture conditions: (i) surrounded by their cumulus and corona cells or (ii) totally denuded. We performed three different studies on sibling oocytes obtained after a standardized buserelin/human menopausal gonadotrophin (HMG) protocol. We investigated the effect of early (1-2 h after retrieval) and late (5-6 h after retrieval) oocyte denudation and injection on the survival and fertilization of the injected oocytes and on embryo cleavage after fertilization. We found no statistically significant differences between early and late injection, indicating that after a standardized buserelin/HMG protocol the metaphase-II oocytes do not need time for further cytoplasmic maturation. Furthermore, a different timing of cumulus-corona cell removal has no effect on the outcome of ICSI, suggesting that the surrounding cells are not necessary for survival, fertilization and cleavage after ICSI. PMID- 9853876 TI - The effect of multiple cryopreservation procedures and blastomere biopsy on the in-vitro development of mouse embryos. AB - Preimplantation genetic diagnosis is currently used in clinical practice. In experienced hands the biopsy procedure alone does not affect the further in-vitro and in-vivo developmental potential of animal and human embryos. No data exist on the combination of cryopreservation of embryos at the pronuclear and/or 8-cell stage and/or biopsy at the 8-cell stage. Pronuclear stages of mouse F1 hybrids (C57Bl/jxCBA) were harvested and divided into several experimental groups. The developmental rates of zygotes, which were neither biopsied nor cryopreserved were used as data control. Others were only cryopreserved at the pronuclear stage (C-PN), or at the cleavage stage (C-CS), or both. Each of these groups was also combined with or without a biopsy. Only the hatched blastocyst rate (HBR), but not the 'simple' blastocyst rate, showed significant differences between groups. Neither C-PN (HBR = 60.42%), nor C-CS (63.16%), nor a combination of both (59.46%) had an impact on the hatched blastocyst rate when compared with that of the control group (67.46%). The biopsy procedure (55.93%) also proved not to be harmful for the embryos. The embryos, which were C-PN and C-CS, and subsequently biopsied, showed a significantly lower hatched blastocyst rate (39.62%) than that of the control, C-PN, C-CS, and C-PN/C-CS groups (P < 0.05). The combination of C PN and cleavage-stage biopsy also lead to a lower hatched blastocyst rate (42.22%), compared with that of the control group (P < 0.05). It was concluded that couples must be advised that an effect on embryos which have undergone a combined cryopreservation and micromanipulation procedure cannot be ruled out. However, cryopreservation at the pronuclear or at the 8-cell stage alone, or in combination with a biopsy procedure, does not influence the further development of the embryo. PMID- 9853877 TI - Successful preimplantation genetic diagnosis is related to the number of available cumulus-oocyte complexes. AB - The inheritance pattern of monogenic inheritable disorders influences the proportion of unaffected embryos after preimplantation genetic diagnosis (PGD). We aimed to investigate the influence of the number of cumulus-oocyte complexes (COC) on the outcome after PGD. Eighty-four cycles of 47 couples were included in our analysis. All couples were at risk of transmitting autosomal recessive, autosomal dominant, X-linked single gene disorders or sexaneuploidies to their offspring. One PGD cycle was carried out for a Yq-deletion of the man. The correlation between the numbers of COC and biopsied embryos and between the numbers of COC and unaffected embryos was highly significant (P <0.05). A pregnancy occurred in 15 cycles and a minimum of six COC were needed to achieve a pregnancy. Thirteen pregnancies were observed in cycles with at least 9 COC. The transfer rate and number of transferred embryos per cycle in the subgroups with <9 COC and > or =9 COC were significantly higher in the latter. Although pregnancy rates did not differ significantly between the two subgroups (probably due to the low number of pregnancies), our data indicate that it is justifiable to cancel PGD cycles in which it is expected that <6 COC will be retrieved and that the couple should be informed about the poor prognosis if <9 COC are retrieved. PMID- 9853878 TI - The receptive endometrium is characterized by apoptosis in the glands. AB - Apoptosis in the human endometrium up to now has been detected during the mid to late luteal phase and therefore connected to the onset of the menstrual shedding. However, there is increasing evidence that regulated apoptosis may be important during decidualization and implantation. To investigate a possible role for apoptosis in the human endometrium and its regulation, we correlated the immunolocalization of the apoptosis regulatory protein bcl-2 and the proliferation marker Ki67 to the in-situ nuclear DNA fragmentation - a key feature of apoptosis - detected by using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labelling (TUNEL) method during the menstrual cycle. Whereas proliferation and bcl-2-expression were predominantly detected in the glandular compartment during the proliferative phase, only single apoptotic cells could be shown during this period. During the transformation of the endometrium (days 15-19) proliferation and bcl-2 expression decreased markedly and there was no sign of apoptosis. At the beginning of the implantation window (days 19-20) we could detect the first signs of apoptosis in the glandular epithelia in the basalis, which extended to the functionalis during the luteal phase. Proliferation and bcl-2 expression are limited to the stromal compartment comprising the large granular lymphocytes - during this time, and extend in parallel with apoptosis from the basal to the functional layers. Apoptosis therefore may be related to the loss of the protective effect of bcl-2 and may have significance for the establishment of an endometrium adequately prepared for successful implantation. PMID- 9853880 TI - Endometrial protein PP14 and CA-125 in recurrent miscarriage patients; correlation with pregnancy outcome. AB - The concentrations of endometrial proteins PP14 and CA-125 were measured in uterine flushings taken on days LH+10 and LH+12 (10 and 12 days after luteinizing hormone surge) of the menstrual cycle from 15 normal, fertile women and 49 women who suffered recurrent miscarriage. The concentration of PP14 was significantly lower in the flushings from the recurrent miscarriage patients than in those from fertile controls on both day LH+10 (median: 1300, range: 3-10 300 ng/ml versus median: 13 933, range: 2174-40 404 ng/ml; P < 0.01) and LH+12 (median: 1560, range: 820-12 100 ng/ml versus median: 14 047, range 1402-62 108 ng/ml; P < 0.05). Similarly concentrations of CA-125 were significantly lower in flushings from recurrent miscarriage women compared to controls on both day LH + 10 (median: 1555, range: 47-6710 U/ml versus median: 6385.5, range 2884-27 731 U/ml, P < 0.01) and LH+12 (median: 2892, range: 956-9974 U/ml versus median: 7127.5, range: 1591-21 343 U/ml; P < 0.05). In contrast there was no significant difference in the concentration of PP14 in plasma samples taken on the same days as the flushings from recurrent miscarriage patients and fertile controls. The concentrations of PP14 in uterine flushings obtained on day LH + 10 or LH + 12 from recurrent miscarriage women during a pre-pregnancy investigative cycle were significantly lower (P < 0.05) in patients who went on to miscarry (median: 1000, range: 9-2900 ng/ml) than those who went on to have a live birth (median: 1440, range: 4-12 100 ng/ml) during a subsequent pregnancy. In contrast there was no significant difference in uterine CA-125 or plasma PP14 concentrations between these two groups of recurrent miscarriage patients. The results suggest that measurements of uterine PP14 and CA-125 may be useful in the assessment of endometrial development in recurrent miscarriage patients and suggest the importance of PP14 in preparing the endometrium for embryo implantation. In addition pre-pregnancy uterine PP14 measurements may be useful in predicting subsequent pregnancy outcome. PMID- 9853879 TI - Precise measurements of intrauterine vascular structures at hysteroscopy in menorrhagia and during Norplant use. AB - Using currently available equipment for panoramic hysteroscopy, the size of images viewed cannot be accurately judged because of the magnifying and distorting effects of the objective lens. This study has demonstrated that magnification by the hysteroscope lens can alter the apparent size of images by up to 27%. An additional effect of lens distortion can alter the apparent size of images viewed by up to 28%, depending on the position of the image in the field of view. These effects are independent and may be additive. Thus, the apparent size of intrauterine structures at hysteroscopy may bear little resemblance to their actual size. Image-correction methods are described which reduce the effects of image magnification on the apparent size of an object viewed through the hysteroscope to 7%, and the effect of distortion to 3 %. This technique can greatly improve the accuracy of measurement at hysteroscopy, and has been utilized in this study for the precise measurement of superficial endometrial vascular diameter (mean+/-SEM) in 34 Norplant users (120+/-11.6 microm) and 20 women with spontaneous menorrhagia (74+/-7.2 microm). It has also confirmed the presence of scattered dilated vessels (up to 777 microm in diameter) on the endometrial surface in some Norplant users. PMID- 9853881 TI - Histological evaluation of endometrium on the day of oocyte retrieval after gonadotrophin-releasing hormone agonist-follicle stimulating hormone ovulation induction for in-vitro fertilization. AB - The objective of this study was to evaluate the histopathological characteristics of endometrial biopsies taken on the day of oocyte recovery in in-vitro fertilization (IVF) cycles with a satisfactory response to ovulation induction. A group of 33 patients who went through ovulation induction for IVF, and in whom an endometrial polyp was suspected on transvaginal ultrasonography during the monitoring phase, were studied. Following oocyte recovery, hysteroscopy, polypectomy and endometrial curettage were performed. Dating of endometrial glands and stroma was carried out in the tissue not containing the polyps. The total dose of follicle stimulating hormone (FSH), duration of ovulation induction, peak oestradiol and luteinizing hormone (LH) concentrations, thickness of endometrium and number of oocytes were recorded and compared to the endometrial dating of the specimens. In 15 cycles (45.5%), the endometrium was classified as 'in phase' (group I), 'advanced' by 2-4 days in a further 15 (45.5%, group II), and in the remaining three cycles (9%) it was delayed in maturation (group III). Younger age was correlated with advanced staging of the endometrium (r = -0.42; P = 0.015). Women with 'in phase' and 'advanced' maturation were similar in their response to ovulation induction; however, there was a strong correlation between advanced dating of endometrium and number of oocytes retrieved (r = 0.49; P = 0.04). Endometrial staging on the day of oocyte retrieval varied widely in patients treated by the same gonadotrophin-releasing hormone agonist (GnRHa)/FSH protocol for ovulation induction. This difference was not predictable by parameters monitored through the cycles. PMID- 9853882 TI - Serum concentrations of soluble human leukocyte class I antigens and of the soluble intercellular adhesion molecule-1 in endometriosis: relationship with stage and non-pigmented peritoneal lesions. AB - Serum concentrations of soluble human leukocyte class I antigens (sHLA-I) and of the intercellular adhesion molecule-1 (sICAM-1) are increased in the early inflammatory stages of several immune-related diseases. These soluble molecules also exert immunomodulatory activity, including regulation of natural killer (NK) cell cytotoxicity. The aim of this study was to verify whether sHLA-I and sICAM-1 serum concentrations are related to the various stages of pelvic endometriosis, which is an immune-related disorder associated with impaired in-vitro NK cell activity. Serum sHLA-I and sICAM-1 concentrations were similar in patients and in healthy donors. However, when evaluated according to disease stage, sHLA-I and sICAM-1 concentrations were higher in patients with endometriosis stage I-II (revised American Fertility Society classification), or with non-pigmented peritoneal lesions. In conclusion, studies on sHLA-I and sICAM-1 may help to clarify the pathogenic mechanisms of endometriosis, and their serum concentrations may serve as additional markers for the early detection of recurrence of the disease during the monitoring of treatment outcome. PMID- 9853883 TI - Interactions between trophoblast and uterine epithelium: monitoring of adhesive forces. AB - At embryo implantation, it is postulated that the initial contact between blastocyst and maternal tissues is by adhesion of the trophoblast to the uterine epithelium. This cell-to-cell interaction is thought to be critical for implantation, although the actual adhesive forces have never been determined. In the present study, the atomic force microscope (AFM) was used to study the adhesion between human uterine epithelial cell lines (HEC-1-A; RL95-2) and human trophoblast-type cells (JAR). Specific interaction forces of these epithelia via their apical cell poles were determined on the basis of approach-and-separation cycles. For this purpose, the AFM tip was functionalized with JAR cells, then brought to the surface of uterine epithelial monolayers and was kept in contact for different periods of time (ms, 1, 10, 20, 40 min). The approach force curves displayed repulsive interactions for both HEC-1-A and RL95-2 cells. However, RL95 2 cells (with a smooth surface structure and a thin glycocalyx) showed lower values of the repulsive regime than HEC-1-A cells (with a rough surface structure and a thick glycocalyx). After having overcome repulsive interactions, the initial contact was followed by adhesive interactions. For contact times of 20 and 40 min, RL95-2 cells, but not HEC-1-A cells, showed specific JAR binding, i.e. the separation force curves displayed repeated rupture events in the range of 1-3 nN with a distance between 7-15 microm and, thereafter, a final rupture event at a distance of up to 45 microm. These features point to the formation of strong cell-to-cell bonds. Collectively, these studies provide the first definition of interaction forces between the trophoblast and the uterine epithelium, and are consistent with the hypothesis that an RL95-2-like architecture of uterine epithelial cells, i.e. an non-polarized phenotype, is essential for apical adhesiveness for the human trophoblast. PMID- 9853884 TI - The accuracy of single serum progesterone measurement in the diagnosis of ectopic pregnancy: a meta-analysis. AB - Serum progesterone measurement has been advocated as a diagnostic tool in the non invasive diagnosis of ectopic pregnancy. To assess the accuracy of a single serum progesterone measurement in the diagnosis of ectopic pregnancy, a meta-analysis was performed incorporating 26 studies evaluating the performance of single serum progesterone measurement in the diagnosis of ectopic pregnancy. A distinction was made in the diagnosis of pregnancy failure of any type versus viable intrauterine pregnancy and the diagnosis of ectopic pregnancy versus non-ectopic pregnancy. The reported sensitivity and specificity differed between the studies. Since there was a clear negative correlation between sensitivity and specificity, summary receiver-operating characteristic (ROC) curves could be estimated. The ROC curve for the diagnosis of pregnancy failure versus viable intrauterine pregnancy showed a good discriminative capacity. Single serum progesterone measurement could not discriminate between ectopic pregnancy and non-ectopic pregnancy. It is concluded that serum progesterone measurement can identify patients at risk for ectopic pregnancy, who need further evaluation, but its discriminative capacity is insufficient to diagnose ectopic pregnancy with certainty. PMID- 9853885 TI - Obstetric outcome of teenage pregnancies. AB - A retrospective cohort study was performed in a tertiary centre to determine if teenage nulliparas (aged alpha19 years, study group) had higher incidences of instrumental and Caesarean deliveries compared with nulliparas aged 20-34 years (control group) selected from the first women in the birth registry who delivered after each study case and satisfying the criteria for controls. The hospital records of the study and control cases were retrieved for review. Comparison was made in the maternal demographics, major antenatal complications, outcome of labour, mode of delivery, and perinatal outcome. In the study group, maternal height was similar but the body mass index was lower. Although the mean birthweight was lower and the incidences of preterm labour and small-for gestational-age infants higher, there were also increased incidences of large-for gestational-age and macrosomic infants. While there was no difference in the types of labour, there were fewer Caesarean and instrumental deliveries, a finding that persisted even after excluding the preterm deliveries. Lastly, teenage mothers aged <17 years had similar outcomes to those aged 17-19 years. These results indicated that teenage mothers had better obstetric outcomes, despite the higher incidence of preterm labour, and that young adolescents (<17 years) performed as well as their older peers. PMID- 9853886 TI - Back pain in in-vitro fertilized and spontaneous pregnancies. AB - The influence of ovarian stimulation in in-vitro fertilization (IVF) on the prevalence of back pain with onset during pregnancy was studied in 31 women who became pregnant after IVF treatment and compared with that of 200 spontaneously pregnant women. A two times higher prevalence rate of sacral pain in late pregnancy was reported among IVF pregnant women (P < 0.0001), as well as a significantly higher prevalence rate of positive results of pelvic pain provocation tests performed in late pregnancy (0.0001 < or = P < or = 0.015), as compared with that of the spontaneously pregnant women. Among the IVF pregnant women, there was a significant positive correlation between relaxin concentrations in early pregnancy and the outcome of pelvic pain provocation tests (0.44 < or = r < or = 0.51, P < 0.05). In addition, the serum relaxin concentration was the factor that best explained differences in sacral pain prevalence. When the influence of serum relaxin concentration on back pain prevalence was taken into account, women carrying multiple pregnancies had no more pain than women carrying singletons, and IVF pregnant women had no more pain than spontaneously pregnant women. These results support the hypothesis that relaxin is involved in the generation of pelvic pain in pregnant women. PMID- 9853887 TI - Randomized trial of conservative laparoscopic treatment and methotrexate administration in ectopic pregnancy and subsequent fertility. AB - Methotrexate treatment was compared to laparoscopic salpingotomy for conservative management of ectopic pregnancy in a prospective randomized study. One hundred patients were randomized into two groups using random numbers. Inclusion criteria were an ectopic pregnancy visualized by ultrasound with a pre-therapeutic score <13 as assessed by the following six criteria, graded from 1 to 3: gestational age, human chorionic gonadotrophin (HCG) concentration, progesterone concentration, abdominal pain, haemoperitoneal volume and diameter of the haematosalpinx. The treatments were either 1 mg/kg of methotrexate injected transvaginally into the ectopic pregnancy without anaesthesia or administered i.m. when the pregnancy could not safely or easily be punctured (group 1), or laparoscopic salpingotomy (group 2). Success was defined as the return to normal (<10 mIU/ml) of HCG concentrations. Treatment was successful for 45 of 51 patients in group 1 (88.2%) and 47 of 49 in group 2 (95.9%). Medical treatment was significantly (P < 0.05) associated with shorter postoperative stay (24 compared with 46 h), but HCG returned to normal more quickly after laparoscopic treatment (13 compared with 29 days). Spontaneous reproductive performance was similar in both groups, but overall intrauterine pregnancy was higher, and repeat ectopic pregnancy lower, after methotrexate treatment. In selected cases of ectopic pregnancy, with a pre-therapeutic score <13, methotrexate treatment appeared as safe and efficient as conservative treatment by laparoscopy and was associated with improved subsequent fertility. PMID- 9853888 TI - A preliminary study of parental stress and child behaviour in families with twins conceived by in-vitro fertilization. AB - This paper reports on a preliminary study of parenting quality, parental stress and child behaviour in families with twins conceived by in-vitro fertilization (IVF) in comparison with families with naturally conceived twins. No differences were found between the types of family in parenting quality or child behaviour. However, parents whose children were conceived by IVF reported greater stress associated with parenting than parents with naturally conceived twins. PMID- 9853889 TI - Pregnancy following intracytoplasmic sperm injection from an HIV-1-seropositive man. AB - The first pregnancy achieved in a seronegative woman following in-vitro fecundation through intracytoplasmic sperm (ICSI) injection from a man with autoimmune deficiency syndrome (AIDS; HIV-1 carrier) is reported. The semen was prepared by PureSperm and swim-up techniques. Some of the motile spermatozoa obtained were used to detect the presence of HIV-1 using the polymerase chain reaction technique. HIV-1 in DNA or RNA form was not detected using this technique. The remaining spermatozoa were frozen. Ovarian stimulation in the woman was performed with long-protocol analogues and gonadotrophins. Thirteen mature oocytes were recovered, into which the thawed spermatozoa were microinjected. Nine embryos were obtained. Four were frozen, four transferred and one discarded. The woman became pregnant. Analyses for HIV-1 in the woman, performed in the first and third months of pregnancy, gave negative results. This case provides further experience with washed semen of sufficient quality for performing artificial insemination in HIV-1-serodiscordant couples (101 inseminations, 31 pregnancies, 28 deliveries, 37 babies, all healthy). In women with obstructed Fallopian tubes, or when the semen is not of sufficient quality for artificial insemination techniques to be performed, ICSI can be carried out using frozen, HIV-1-free semen. PMID- 9853890 TI - Do men undergoing sterilizing cancer treatments have a fertile future? AB - This study was designed to assess the effect of cancer treatments on the natural and assisted reproductive potential of men. A cohort of men with cancer, in whom radiotherapy and/or chemotherapy was planned, were invited to participate. Twenty two pre- and post-treatment semen samples were analysed. The reproductive potential of participants was assessed with respect to the current range of fertility treatment options available. Abnormal sperm concentrations were found in 27% of patients pre-treatment compared to 68% post-treatment following a mean latency of 20 months from treatment. Fifty-nine percent of patients experienced a clinically significant decrease in sperm, concentration following radiotherapy and/or chemotherapy; 23% developed azoospermia following treatment. Eighty-two percent of patients with testicular malignancy had oligo- or azoospermia post treatment. Only one patient had a clinically significant reduction in the percentage of motile spermatozoa post-treatment. Cryopreservation of semen prior to treatment improved the fertility prospects of 55% of patients. Intracytoplasmic sperm injection (ICSI) enhanced the fertility prospects of a further 14%. In the absence of, or after depletion of, cryopreserved semen, ICSI could enhance the fertility prospects of 45% of patients. Fertilization has been achieved by ICSI using spermatozoa retrieved by testicular biopsy from an azoospermic testicular cancer survivor 8 years after chemotherapy. It was concluded that chemotherapy and/or radiotherapy may depress semen concentration to the extent of rendering a man infertile. The severity of the reduction in sperm concentration following treatment is unpredictable but likely to be most severe in those with testicular malignancy and those treated with radiotherapy or alkylating chemotherapy agents. Not all men are keen to undergo an appraisal of their post-treatment fertility potential, for reasons which are unclear. Improving awareness and education of patients concerning the effects of both cancer and cancer treatments on reproductive potential is essential. With the advent of ICSI, it is possible to offer a very reasonable chance of conception in all men with cancer who present for cryopreservation of semen prior to treatment in whom spermatozoa (even in very low concentrations) are present in the ejaculate. PMID- 9853891 TI - A programme of semen cryopreservation for patients with malignant disease in a tertiary infertility centre: lessons from 8 years' experience. AB - The improved survival in recent years of young males suffering from cancer, and an understanding of the gonadotoxic effects of chemotherapy treatment, have motivated patients and clinicians to preserve fertility potential before embarking on adjuvant therapy. Among 231 men (mean age 28.0; range 15-56 years) diagnosed with malignant disease and referred to our unit for semen cryopreservation, 112 patients (49.8%) had reduced sperm quality of <10 x 10(6) motile spermatozoa per ejaculate; however, most had sufficient suitable spermatozoa for freezing. In 40 patients (17.3 %) the semen samples were not frozen because of complete azoospermia (n = 32) or only immotile sperm in the ejaculate (n = 2), while six men were unable to produce a single sample. Some 79 men had testicular tumours (group I), 121 suffered from haematological malignancy (leukaemia or lymphoma; group II), and 27 had cancer of different causes (group III). Men in group I had significantly lower (P < 0.001) sperm quality compared with groups II and III. There was no difference between patients with seminoma and non-seminoma tumours. In the haematological malignancy group there was no difference in sperm parameters between leukaemia (n = 12) and lymphoma (n = 77) patients, but patients with Hodgkin's lymphoma had significantly lower sperm quality compared with non-Hodgkin's lymphoma. Following chemotherapy, six couples attended the clinic for assisted conception treatment using the frozen semen. Two had successful intrauterine insemination cycles which each resulted in delivery of a healthy girl; one couple had conceived in their first in-vitro fertilization (IVF) attempt, followed by delivery of healthy twins. Two women conceived after intracytoplasmic sperm injection treatment and the sixth woman achieved only biochemical pregnancy after numerous IVF and frozen embryo replacement cycles. We recommend that a properly designed programme for semen cryopreservation for cancer patients should be developed in leading tertiary assisted conception centres, which have adequate facilities and experience for cryopreservation and can offer the whole range of appropriate assisted reproductive treatment and counselling. PMID- 9853892 TI - Psychological reactions during in-vitro fertilization: similar response pattern in husbands and wives. AB - The purpose of this study was to examine differences in daily emotional, physical and social reactions among husbands and wives during in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Forty couples about to undergo ICSI or IVF at a private infertility clinic monitored their emotional, physical and social reactions daily for one complete treatment cycle from the first day of stimulation until the outcome of treatment was known (approximately 35 days). The results showed that men and women had a similar response pattern to oocyte retrieval, fertilization, embryo transfer and the pregnancy test. These stages were associated with the most significant changes in reactions for both spouses. The pattern of results suggested that the most important psychological determinant of reactions during IVF was the uncertainty of treatment procedures. Spouses appeared to be equally sensitive to this uncertainty and both appeared to respond to it with ambivalent feelings involving emotional distress and positive feelings of hope and intimacy. PMID- 9853893 TI - The management of genetic origins: secrecy and openness in donor assisted conception in Israel and elsewhere. AB - Donor assisted conception provides new opportunities for achieving parenthood but at the same time raises issues of secrecy, anonymity, and the management of the offspring's genetic origins. As with adoption, the child's right to a genetic identity is at stake. The first part of this paper examines bio-psychosocial evidence indicating that maintaining secrecy and anonymity regarding genetic parents may be detrimental to the well-being of the donor offspring, the donor, and the nurturing, social parents. The second part discusses different approaches to the disclosure of genetic origins. The third part presents the case of Israel, as reflected mainly in the Report of the Israeli Ministry of Justice (1994). The paper shows that the Committee, yielding to the medical profession in the country, preferred the interests of the adults involved in donor assisted conception to those of the offspring and made no sincere attempts to protect the child's basic right to genetic identity. In doing so, it turned its back on the well established policy of openness in the field of adoption and on the emerging trend to similar openness in donor assisted conception in other countries. PMID- 9853894 TI - Sharing genetic origins information in donor assisted conception: views from licensed centres on HFEA donor information form (91) 4. AB - Section 31 of the Human Fertilization and Embryology Act 1990 permits people born as a result of licensed treatments provided in the UK to seek certain information about their genetic origins held on the Human Fertilization and Embryology Authority (HFEA) Register of Information. The precise nature of such information is to be specified in Regulations that have yet to be determined by the Secretary of State. The Register comprises data submitted to the Authority by licensed centres on HFEA Donor Information Form (91) 4. This paper reports on a survey of views of licensed centres concerning the adequacy of this form and centres' experiences of gathering information from donors. The survey shows that centres' experiences vary considerably, and the authors argue that there is a case for the sharing of centres' experiences and dissemination of effective strategies. The paper further discusses the nature of information that might become available to individuals seeking information from the HFEA. While requests for non-identifying information concerning the donor cannot be made prior to 2010, any information made available then is inevitably dependent on current practice in centres. The authors conclude that current variability in practice does not best serve the interests of donor offspring who may seek information about their genetic origins. PMID- 9853895 TI - Temporal trends in twinning rates in Italy around World War II. PMID- 9853896 TI - Time of insemination within the cycle and offspring sex ratio. PMID- 9853897 TI - Leptin concentrations in stimulated cycles. PMID- 9853898 TI - Contrast-enhanced sonography. PMID- 9853899 TI - Previous abortions and risk of pelvic endometriosis. PMID- 9853900 TI - Synaptic organisation of lumbar sympathetic ganglia of guinea pigs: serial section ultrastructural analysis of dye-filled sympathetic final motor neurons. AB - The authors serially sectioned seven dye-filled neuronal somata and more than 1.6 mm of their dendrites from the lumbar sympathetic ganglia of guinea pigs and examined them ultrastructurally to determine the distribution of preganglionic synaptic inputs to their dendrites and cell bodies. Most of the surface of the neurons was covered with Schwann cells. Apposing boutons were rare, with an average density of one axosomatic bouton per 125 microm2 of somatic membrane and one axodendritic bouton per 25 microm of dendrite. Many dendritic segments that were more than 50 microm long completely lacked any apposing boutons. Although the average density of apposing boutons was low, local densities could be high, so that clusters of up to four adjacent boutons occurred on cell bodies and dendrites alike. The spatial arrangement of the apposing boutons for each of the cells examined here was not significantly different from a random distribution. Consequently, the number of apposing boutons observed for any neuron was simply proportional to the amount of neuronal surface sampled in the serial section run. About 50% of boutons directly apposing the neurons lacked any detectable presynaptic specialisations. When they were present, the presynaptic densities had a mean length of about 220 nm, with no difference between boutons that made axosomatic or axodendritic appositions. By applying these data to complete reconstructions of the dendritic trees of dye-filled sympathetic neurons at the light microscopic level, the authors estimated that few neurons in the lumbar sympathetic chain of guinea pigs would receive more than 200 synapses or apposing boutons and that many of them would receive less than 100 synapses. Up to 50% of these boutons would be predicted to make axosomatic contacts. These new observations provide a strong morphological framework for a better understanding of how sympathetic final motor neurons process their preganglionic synaptic inputs. PMID- 9853901 TI - Prominent expression of glial cell line-derived neurotrophic factor in human skeletal muscle. AB - Glial cell line-derived neurotrophic factor (GDNF) has been shown to exert neurotrophic effects on motor neurons as well as mesencephalic dopaminergic neurons. Because GDNF promotes survival of motor neurons in vivo and in vitro and rescues motor neurons from naturally occurring cell death, the potential use of GDNF for treatment of motor neuron diseases has been a major focus of recent research. The expression of GDNF in humans, however, has not been fully examined. In the present study, we examined the expression of GDNF in adult human muscle by Northern blot, reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemical analyses to address physiological roles of GDNF in humans. Northern blot analysis demonstrated high expression of GDNF mRNA in human skeletal muscle when compared to that of mouse. Intense GDNF immunoreactivity was observed in the vicinity of plasma membranes of skeletal muscle, particularly at neuromuscular junctions. GDNF immunoreactivity was also observed within the axons and surrounding Schwann cells of peripheral nerves. However, RT-PCR detected expression of GDNF mRNA only in skeletal muscle, and not within the anterior horn cells of human spinal cord. These results suggest that GDNF is produced by skeletal muscle and taken up at the nerve terminals for retrograde transport by axons. Thus, GDNF in human skeletal muscle may be involved in promoting motor neuron survival as a target-derived neurotrophic factor. PMID- 9853902 TI - Thalamic territories innervated by cerebellar nuclear afferents in the hedgehog tenrec, Echinops telfairi. AB - To gain more insight into the evolution and functional significance of cerebrocerebellar circuits, the cerebellothalamic projections were studied with anterograde tracer substances in the Madagascan lesser hedgehog, tenrec. This insectivore shows one of the lowest size indices among mammals for both the cerebellar nuclei and the neocortex. Almost all cerebellodiencephalic target areas found in the tenrec have been described in other mammals. The intensity and extent of particular projections, however, vary considerably in the tenrec compared with the other mammals investigated so far. The most remarkable finding may be the tenrec's cerebellar projection to the nucleus ventralis medialis. This projection is the most prominent cerebellothalamic projection and originates in predominantly the lateral portion of the cerebellar nuclear complex. The projection to the caudolateral portion of the ventralis anterior complex (VAC) is located immediately rostral to the area receiving ascending somatosensory afferents and appears to originate, in particular, from the intermediate cerebellar nuclear complex. Another cerebellothalamic focus of terminations lies in the paralamellar region of the VAC, whereas the proper intralaminar nuclei, at best, receive a sparse cerebellar input. A faint-to-moderate projection, on the other hand, has been traced consistently to the ventral portion of the lateralis posterior-pulvinar complex and the adjacent dorsal geniculate nucleus. In addition, there are prominent cerebellosubthalamic projections to the zona incerta and the ventral geniculate nucleus. The latter projection is confined mainly to the ventralmost subdivision, which has been shown previously to receive ascending somatosensory, but not retinal, afferents. With the exception of the nucleus ventralis medialis, the projections were essentially confined to the contralateral side. PMID- 9853903 TI - Superior area 6 afferents from the superior parietal lobule in the macaque monkey. AB - Superior area 6 of the macaque monkey frontal cortex is formed by two cytoarchitectonic areas: F2 and F7. In the present experiment, we studied the input from the superior parietal lobule (SPL) to these areas by injecting retrograde neural tracers into restricted parts of F2 and F7. Additional injections of retrograde tracers were made into the spinal cord to define the origin of corticospinal projections from the SPL. The results are as follows: 1) The part of F2 located around the superior precentral dimple (F2 dimple region) receives its main input from areas PEc and PEip (PE intraparietal, the rostral part of area PEa of Pandya and Seltzer, [1982] J. Comp. Neurol. 204:196-210). Area PEip was defined as that part of area PEa that is the source of corticospinal projections. 2) The ventrorostral part of F2 is the target of strong projections from the medial intraparietal area (area MIP) and from the dorsal part of the anterior wall of the parietooccipital sulcus (area V6A). 3) The ventral and caudal parts of F7 receive their main parietal input from the cytoarchitectonic area PGm of the SPL and from the posterior cingulate cortex. 4) The dorsorostral part of F7, which is also known as the supplementary eye field, is not a target of the SPL, but it receives mostly afferents from the inferior parietal lobule and from the temporal cortex. It is concluded that at least three separate parietofrontal circuits link the superior parietal lobule with the superior area 6. Considering the functional properties of the areas that form these circuits, it is proposed that the PEc/PEip-F2 dimple region circuit is involved in controlling movements on the basis of somatosensory information, which is the traditional role proposed for the whole dorsal premotor cortex. The two remaining circuits appear to be involved in different aspects of visuomotor transformations. PMID- 9853904 TI - Differential regional and cellular distribution of dopamine D2-like receptors: an immunocytochemical study of subtype-specific antibodies in rat and human brain. AB - Dopamine D2-like receptors (D2, D3, and D4) are major targets for action of typical and atypical neuroleptics, commonly used in the treatment of schizophrenia. To understand their individual functional contribution, subtype selective anti-peptide antibodies were raised against D2, D3, and D4 receptor proteins. The antibodies were shown to be specific on immunoblots of rat brain membranes and immunoprecipitated the solubilized native dopamine receptors in an antibody concentration-dependent manner. In addition, they also bind selectively to the respective recombinant D2, D3, and D4 receptor membrane proteins from cDNA transfected cells. Immunolocalization studies show that the D2-like receptor proteins had differential regional and cellular distribution in the cerebral cortex, hippocampus, basal ganglia, cerebellum, and midbrain, thus providing anatomical substrate for area-specific regulation of the dopamine neurotransmission. In cortical neurons, D4 receptor protein was found in both pyramidal and nonpyramidal cells, whereas D2 and D3 seem to be mostly associated with nonpyramidal interneurons. In rat hippocampus, the expression pattern of D2 like receptors (D4>D3>D2) mirrored that obtained with immunoprecipitation studies. D2 and D4 receptor immunolabeling was observed in the thalamic reticular nucleus, which was negative for the D3 subtype. Species differences were also observed; for example, the D4 subtype receptor is the most highly expressed protein in the rat cortex, whereas it is significantly less in human cortex. Differential patterns of D2, D3, and D4 receptor expression in rat and human brain should shed light on the therapeutic actions of neuroleptic drugs and may lead to the development of more specifically targeted antipsychotic drugs. PMID- 9853905 TI - Species differences in the expression and distribution of the neuropeptide Y Y1, Y2, Y4, and Y5 receptors in rodents, guinea pig, and primates brains. AB - The respective distribution of neuropeptide Y Y1, Y2, Y4, and Y5 receptor subtypes was investigated in rodents (rat and mouse), guinea pig, and primates (marmoset and vervet monkeys and human) brains, representing three orders of mammals. [125I] [Leu31,Pro34]PYY (total Y1-like; Y1, Y4, and Y5) and [125I]PYY3 36 (total Y2-like; Y2 and possibly Y5) binding sites were discretely but similarly distributed in the rat and mouse brain, each having its unique pattern. In contrast, surprisingly low levels of [125I]PYY3-36 binding sites were found in the guinea pig brain including in the hippocampal formation. [125I][Leu31,Pro34]PYY/BIBP3226-insensitive binding sites (Y5-like) were found in different areas of the rat and guinea pig brains. The primate brains also revealed a different distribution binding profile for these various NPY receptor subtypes. Although the human and vervet brains contained very low amounts of [125I][Leu31,Pro34]PYY sites (Y1-like) in most brain regions, except for the dentate gyrus of the hippocampus, the marmoset brain contains significant amounts of both [125I][Leu31,Pro34]PYY (Y1-like) and [125I]PYY3-36 (Y2-like) binding sites. Additionally, [125I][Leu31,Pro34]PYY/BIBP3226-insensitive binding sites were not clearly detected in the vervet and human brains. On the other hand, Y5 like binding sites were observed in few regions of the marmoset brain. Finally, [125I]hPP (Y4/Y5-like) were very discretely distributed in the rat brain, being concentrated in the paraventricular nucleus of the hypothalamus and the interpeduncular nucleus. The marmoset brain is apparently not enriched with specific [125I]hPP sites. Taken together, these data show that significant species differences exist in the level of expression and distribution of various NPY receptor subtypes in the mammalian brain. PMID- 9853906 TI - Distribution of neurons expressing immunoreactivity for the 5HT3 receptor subtype in the rat brain and spinal cord. AB - The cellular distribution of the type 3 serotonin receptor (5HT3R) in the rat brain was established immunocytochemically by using a polyclonal antibody raised against a synthetic peptide from the deduced amino-acid sequence of the cloned 5HT3R. The 5HT3R-immunoreactive neurons were found in the forebrain, brainstem, and spinal cord, but within each region, the intensity of the immunoreactivity differed considerably. Within the forebrain, intensely immunoreactive cells were found in layers II-III of the neocortex, anterior olfactory nucleus, hippocampal formation, and amygdala. A few strongly immunoreactive neurons were consistently observed in the caudate putamen, and moderately or weakly labeled neurons were occasionally found in the nucleus accumbens. Within the brainstem, intensely labeled neurons were found in the trigeminal motor (V) and facial (VII) nuclei. Immunostained neurons were detected in the dorsal and the ventral horn of the spinal cord. These results reveal that the 5HT3R-immunoreactive neurons are broadly distributed throughout the rat brain and spinal cord, and suggest that this receptor can subserve significant participation in central nervous system neurotransmission. PMID- 9853907 TI - Olfactory bulb development is altered in small-eye (Sey) mice. AB - Small-eye (Sey) is a spontaneous, semidominant murine mutation that results from a point mutation in the Pax-6 gene. Both the eyes and the olfactory system fail to develop in homozygotes and these animals die neonatally. Heterozygotes (Sey/+) have different degrees of eye abnormalities including decreased lens size and cataracts. In the present study, we examined whether one mutated allele of Pax-6 also affects olfactory system development. By 42 days of age, main olfactory bulb volume was significantly decreased in Sey/+ animals compared with wild-type littermates, and this effect was even more dramatic in 70-day-old animals. In contrast, there was no effect on accessory olfactory bulb, olfactory epithelial, or vomeronasal organ development at any age in Sey/+ animals, demonstrating the specificity of the effect. In the main olfactory bulb, the largest differences in laminar volume were found in the glomerular and granule cell layers. These layers contain the olfactory bulb interneurons, and a subpopulation of these cells were found to be Pax-6 immunoreactive. Examination of the neurochemical consequences of this mutation showed that the number of both tyrosine hydroxylase (TH)- and gamma-aminobutyric acid (GABA)-immunoreactive profiles were dramatically decreased in Sey/+ animals as compared with controls. In contrast, neither calretinin nor calbindin immunoreactivity was affected by this mutation. Dual labeling immunohistochemistry showed that nearly all TH-immunoreactive cells and a subpopulation of GABA-immunoreactive cells coexpressed Pax-6. However, calretinin- and calbindin-immunoreactive cells were not Pax-6 immunopositive. These data indicate that two normal alleles of Pax-6 are required for normal olfactory bulb development and, as part of this effect, this gene may be involved in the development of specific neurotransmitter systems. PMID- 9853908 TI - Chemical anatomy of the macaque monkey olfactory bulb: NADPH-diaphorase/nitric oxide synthase activity. AB - The distribution and the morphology of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase (ND)-active and neuronal nitric oxide synthase (NOS) immunoreactive neurons and fibers were studied in the olfactory bulb of three species of primates, i.e., the cynomolgus macaque monkey (Macaca fascicularis), the Japanese macaque monkey (Macaca fuscata), and the pig-tail macaque monkey (Macaca nemestrina). The ND staining was carried out by means of a direct histochemical method with beta-NADPH as cosubstrate and nitro blue tetrazolium as chromogen. The NOS immunostaining was carried out by using a polyclonal antibody and the avidin-biotin peroxidase method. Similar results were found in the three species, where a distinct distribution pattern of ND/NOS-stained neurons and fibers was observed. All olfactory fibers demonstrated ND-positive labeling but they were NOS-immunonegative. In the superficial modulatory area of the olfactory bulb, a few weakly ND- and NOS-positive periglomerular cells, stellate cells, and darkly stained superficial short-axon cells were observed. In the inframitral layers, granule cells, deep stellate cells, and deep short-axon cells were distinguished. Short-axon cells had oriented morphologies and spiny dendrites. Many thick, varicose ND/NOS-stained fibers identified as centrifugal fibers were observed in the white matter, granule cell layer, internal plexiform layer, mitral cell layer, and external plexiform layer. This distribution of ND activity and NOS immunoreactivity showed similarities to and differences from what has been reported in the olfactory bulb of macrosmatic mammals including rodents (rat, mouse, and hamster) and insectivores (hedgehog). These data confirm that the complexity of the ND/NOS staining in the olfactory bulb of one species correlates with the importance of olfaction in the biology of such species. PMID- 9853909 TI - Patient education: a tool in the outpatient management of deep vein thrombosis. AB - A key to effective outpatient management of thromboembolic disease is patient education. Although highly effective for the treatment of deep vein thrombosis (DVT), antithrombotic treatment may fail as a result of inadequate patient education. The risk of hemorrhage from antithrombotic drugs is related to a number of factors including intensity of anticoagulation achieved, comorbid illness, concurrent drug therapy, and lifestyle. When patients receive inadequate antithrombotic treatment, the risk of recurrent thromboembolic events and long term complications are substantially increased. A well-organized, structured education program enables patients to learn the necessary skills that permit complex and valuable therapies to be managed on an outpatient basis. Health care professionals who are part of an outpatient DVT treatment program should possess working knowledge of adult learning theory and instructional design. To be effective, education programs should be systematically planned, have an educationally sound structure, and attempt to meet specific objectives. In addition, they should build on patients' existing knowledge, skills, and attitudes. Periodic evaluation of the education program is important to ensure that overall goals are being adequately met and to identify areas of weakness. PMID- 9853910 TI - Issues in developing an outpatient deep venous thrombosis treatment program: university hospital experience. AB - Low-molecular-weight heparins (LMWHs) are safe and effective in the treatment of venous thromboembolic disease. Since they require less therapeutic monitoring than unfractionated heparin and may be administered to outpatients, it is possible to decrease overall health care costs of patients diagnosed with deep vein thrombosis (DVT) without compromising quality of care. In developing protocols for outpatient management of DVT, treatment components must be analyzed and adapted to meet the needs of populations served by specific health care settings. Issues include medicolegal risk, financial analysis, program coordination, patient selection, product selection, timing, administration strategies, education, and outcomes monitoring. PMID- 9853911 TI - Outpatient deep vein thrombosis treatment models. AB - Approximately 9000 patients are admitted annually to Canadian hospitals with a primary diagnosis of deep vein thrombosis (DVT). Although this is a serious medical condition, potentially more than 40% of all patients with uncomplicated DVT, or 3600 Canadian patients/year, may be safely treated as outpatients with low-molecular-weight heparin. Outpatient treatment avoids costly hospitalization that is required for the standard 5-10 days of intravenous unfractionated heparin therapy. Although institutions vary widely in the available resources, five core models can assist with successful implementation of an outpatient DVT treatment program while providing optimum use of each site's resources and clinical expertise. These models are as follows: anticoagulation clinic (thromboembolic clinic-service), medical day care clinic, emergency department fast-track, one visit and self-injection, and physician-office follow-up. A program was implemented at Burnaby Hospital in May 1996 using the medical day care clinic model as a pilot. Formal evaluation of the program is still in progress but interim evaluation demonstrated overwhelming success. PMID- 9853912 TI - Patient outcomes and cost analysis associated with an outpatient deep venous thrombosis treatment program. AB - Low-molecular-weight heparins are safe and effective for outpatient treatment of deep vein thrombosis (DVT). An outpatient DVT treatment program was established in a 532-bed nonprofit teaching hospital. With the cooperation and skills of a multidisciplinary team, 125 patients were treated for acute DVT primarily at home over 26 months. Preliminary results demonstrated a drop in average length of hospitalization from 5.4 days before the program to 0.97 days. All patients successfully completed a course of enoxaparin, averaging 5.23 days, with 4.26 days of treatment as outpatients. One hundred twenty-two patients (97.6%) required no rehospitalization for any reason, including recurrence of DVT and bleeding complications, throughout the 90-day anticoagulation treatment course. The hospital achieved a cost avoidance of $2470.68/patient. For all 125 patients, total cost avoidance was $308,835.17. PMID- 9853913 TI - Coordinated regulation of genes of the nitric oxide and endothelin pathways during the development of pulmonary hypertension in fetal lambs. AB - Ligation of the ductus arteriosus in utero produces fetal and neonatal pulmonary hypertension and alterations in the hemodynamic responses to nitric oxide and endothelin-1 in fetal and newborn lambs. To determine whether fetal pulmonary hypertension alters the expression of the genes of the nitric oxide and endothelin-1 pathways, seven fetal lambs (123-126-d gestation) underwent ligation of the ductus arteriosus. Near-term (138-139-d gestation), total lung RNA, and protein were prepared from control and ductal ligation fetal lambs for RNase protection assays and Western blotting. Ligation of the ductus arteriosus was associated with decreased expression of endothelial nitric oxide synthase mRNA and protein, and the alpha1 and the beta1 subunits of soluble guanylate cyclase protein; and with increased expression of phosphodiesterase V mRNA. Ligation of the ductus arteriosus was also associated with increased expression of preproendothelin-1 mRNA and with decreased expression of endothelin B receptor (ET(B)) mRNA. These results suggest that there is coordinated regulation of genes of the nitric oxide pathway, which would decrease nitric oxide and cGMP concentration, thereby decreasing pulmonary vasodilator activity. There is also coordinated regulation of genes of the endothelin-1 pathway, which would increase endothelin-1 concentration and limit ET(B) receptor activation, thereby increasing pulmonary vasoconstrictor activity. These alterations in gene expression would increase fetal pulmonary vascular resistance, contributing to the development of pulmonary hypertension after birth. PMID- 9853914 TI - Pulmonary and systemic effects of the phosphodiesterase inhibitor dipyridamole in newborn lambs with persistent pulmonary hypertension. AB - Nitric oxide (NO) relaxes vascular smooth muscle by increasing the intracellular concentration of cGMP. In the pulmonary circulation, cGMP is inactivated by specific phosphodiesterases (PDE5). Dipyridamole, a clinically approved drug, has inhibitory activity against PDE5 and has been reported to augment the response to inhaled NO in persistent pulmonary hypertension of the newborn (PPHN). We wished to determine whether dipyridamole alone, or in combination with NO, can be used to treat a newborn lamb model of PPHN. In newborn lambs with PPHN, dipyridamole infused at 0.02 mg/kg/min for 45 min alone, or in combination with 5 ppm of inhaled NO for the final 15 min, significantly decreased pulmonary and systemic blood pressure, decreased pulmonary vascular resistance, and increased pulmonary blood flow. There was no significant difference between the pulmonary vascular effects of 5 ppm NO alone compared with the effects of NO combined with dipyridamole. In control lambs, the 45-min infusion of dipyridamole did not change pulmonary pressure whereas systemic pressure decreased by 28 +/- 3%. These systemic effects in control lambs persisted 90 min after discontinuing the dipyridamole infusion. Systemic arteries isolated from both control and PPHN lambs were significantly more sensitive to dipyridamole than pulmonary arteries. We conclude that dipyridamole has significant hemodynamic effects in both the pulmonary and systemic circulations of newborn lambs with pulmonary hypertension as well as in the systemic circulation of newborn control lambs. The pronounced effects of dipyridamole on the systemic circulation limits its utility as an adjunct to inhaled NO in the treatment of PPHN. PMID- 9853915 TI - Involvement of thromboxane A2 and prostacyclin in the early pulmonary hypertension after porcine meconium aspiration. AB - Severe perinatal aspiration of meconium is frequently complicated by unsuccessful neonatal adaptation with associated pulmonary hypertension. This vascular complication is supposedly related to pulmonary release of vasoconstrictory agents, including metabolites of arachidonic acid. Thus, to investigate the role of prostanoids on these meconium-induced circulatory changes in the lungs, the hemodynamic response to meconium instillation was studied in acetylsalicylic acid pretreated juvenile pigs. Twelve 10-wk-old pigs with adapted lung circulation received 3 mL/kg of 65 mg/mL human meconium via the endotracheal tube. Six of them were medicated with 10 mg/kg acetylsalicylic acid 30 min before meconium insufflation. Hemodynamic parameters and urinary excretion of stable metabolites of thromboxane A2 and prostacyclin were measured serially for 6 h after the insult. Meconium administration induced a biphasic increase in mean pulmonary artery pressure and pulmonary vascular resistance, and a rapid rise in urinary levels of prostanoid metabolites. Acetylsalicylic acid pretreatment prevented the initial (0-1 h) pulmonary hypertensive response and increase in prostanoid excretion. During the second phase (1-6 h), acetylsalicylic acid did not attenuate the progressive increase in mean pulmonary artery pressure and pulmonary vascular resistance nor did it affect the longitudinal distribution of the pulmonary resistances. Our results thus show that in adapted porcine lungs, arachidonic acid metabolites contribute to the early hypertensive response, but have only minor effects during the second phase vascular hypertension. PMID- 9853916 TI - Pulmonary hemodynamics and plasma endothelin-1 during hypoxemia and reoxygenation with room air or 100% oxygen in a piglet model. AB - The immediate effect on the pulmonary circulation of reoxygenation with either room air or 100% O2 was studied in newborn piglets. Hypoxemia was induced by ventilation with 8% O2 until base excess was <-20 mmol/L or mean arterial blood pressure was <20 mm Hg. Reoxygenation was performed with either room air (n = 9) or 100% O2 (n = 9). Mean pulmonary artery pressure increased during hypoxemia (p = 0.012). After 5 min of reoxygenation, pulmonary artery pressure increased further from 24 +/- 2 mm Hg at the end of hypoxemia to 35 +/- 3 mm Hg (p = 0.0077 versus baseline) in the room air group and from 27 +/- 3 mm Hg at the end of hypoxemia to 30 +/- 2 mm Hg (p = 0.011 versus baseline) in the O2 group (NS between groups). Pulmonary vascular resistance index increased (p = 0.0005) during hypoxemia. During early reoxygenation pulmonary vascular resistance index decreased rapidly to values comparable to baseline within 5 min of reoxygenation in both groups (NS between groups). Plasma endothelin-1 (ET-1) decreased during hypoxemia from 1.5 +/- 0.1 ng/L at baseline to 1.2 +/- 0.1 ng/L at the end of hypoxemia (p = 0.003). After 30 min of reoxygenation plasma ET-1 increased to 1.8 +/- 0.3 and 1.5 +/- 0.2 ng/L in the room air and O2 groups, respectively (p = 0.0077 in each group versus end hypoxemia; NS between groups). We conclude that hypoxemic pulmonary hypertension and plasma ET-1 normalizes as quickly when reoxygenation is performed with room air as with 100% O2 in this hypoxia model with newborn piglets. PMID- 9853917 TI - The Shc 66 and 46 kD isoforms are differentially downregulated at parturition in the fetal mouse lung. AB - Many of the signaling pathways regulating fetal lung mesenchymal cell proliferation are mediated by the Shc intracellular signaling proteins. Shc is expressed as three isoforms: 52 kD and 46 kD proteins (Shc 52 and Shc 46, respectively) translated from the same mRNA, and a 66 kD form (Shc 66) translated from a separate mRNA. Shc 52 is an activator of Ras and mitogen-activated protein kinase, whereas Shc 66 antagonizes Ras activation. The function of Shc 46 is unclear. We hypothesized that the Shc isoforms are differentially regulated during fetal mouse lung morphogenesis. Relative Shc 66 and Shc 46 protein expression are high until parturition (term = 18.5 d), when a dramatic decrease begins; by postconceptual d 20, relative Shc 66 and Shc 46 expression have fallen by 75 and 69%, respectively. A similar pattern of decreasing Shc 66 mRNA expression in the peripartum period was detected by reverse transcription and competitive polymerase chain reaction during the same period. By isoform-specific immunohistochemistry, Shc 66 is widely distributed in the embryonic lung but becomes restricted to the bronchial smooth muscle and overlying epithelia, periarterial smooth muscle, and the interlobar pleura late in gestation. After parturition, Shc 66 is virtually absent from the lung. All three Shc isoforms are phosphorylated by epidermal growth factor stimulation in fetal lung mesenchymal cells, indicating that Shc 66 is functional in these cells. These data indicate that Shc isoforms are differentially regulated during lung development. PMID- 9853918 TI - Surfactant protein-C in ventilated premature lamb lung. AB - Although surfactants containing only lipids and surfactant protein C (SP-C) or SP C analogs can be effective for the treatment of surfactant deficiency in animal models, there is no information concerning the alveolar or lung clearance of SP C. Because the other lipid and protein components of surfactant are cleared very slowly from the preterm lung, we hypothesized that SP-C also would be cleared slowly. Therefore, we compared the losses of iodinated native SP-C (nSP-C) and a recombinant SP-C analog (rSP-C, phenylalanines in positions 4 and 5 and isoleucine in position 32 of the human sequence) to [14C]dipalmitoylphosphatidylcholine (DPPC) after airway administration at birth of trace or treatment doses of surfactant given to preterm lambs. In preterm lambs given trace doses at 134-136 d gestation, alveolar [14C]DPPC and [125I]rSP C decreased to 14.7% recovery for DPPC and 8.3% recovery for rSP-C after 2 h ventilation. There was no loss of [14C]DPPC from the total lungs (alveolar wash + lung tissue), and approximately 20% of the [125I]rSP-C was lost from the lungs. For 128 d gestational age lambs treated with 100-mg/kg doses of surfactants containing nSP-C or 2% rSP-C, the alveolar and total lung recoveries for [125I]nSP-C or [125I]rSP-C were equivalent to that of [14C]DPPC after 5 h ventilation. These results demonstrate that nSP-C and rSP-C have alveolar clearances and accumulations into preterm lung tissue that are similar to those of DPPC. PMID- 9853919 TI - Tissue-specific and developmental regulation of transforming growth factor-beta1 expression in fetal lamb ductus arteriosus endothelial cells. AB - We previously established that increased hyaluronan synthesis in ductus arteriosus (DA) compared with aorta (Ao) endothelial cells (EC) from early gestation fetal lambs (100-d, term = 145 d) is transforming growth factor-beta (TGF-beta)-dependent. We now address whether this is associated with tissue specific and developmentally up-regulated expression of TGF-beta1 in the 100-d DA EC. Immunoprecipitation revealed TGF-beta synthesis doubled in DA versus Ao EC from 100-d gestation lambs (p < 0.05). In 138-d DA EC, TGF-beta protein levels were reduced (p < 0.05) and comparable to those in Ao cells. Western immunoblotting with a beta1 isoform-specific antibody confirmed these differences as being related to TGF-beta1. Northern blot analysis demonstrated that TGF-beta1 mRNA levels were slightly but not significantly increased in 100-d DA compared with Ao EC, despite its short half-life in DA (9.5 h) versus Ao EC (20 h). TGF beta1 mRNA levels were reduced in 138-d DA and Ao EC (p < 0.05), and the mRNA half-life was comparable in DA (9 h) versus Ao (13 h). Nuclear run-on analysis confirmed increased TGF-beta1 mRNA transcription in 100-d DA versus Ao and 138-d DA EC. Thus, up-regulated TGF-beta1 expression in 100-d DA compared with Ao cells is due to increased transcription and translation of a relatively unstable mRNA, and its down-regulation in 138-d DA and Ao EC is related to reduced mRNA transcription and stability, respectively. PMID- 9853920 TI - Serum levels of basic fibroblast growth factor and vascular endothelial growth factor in children and adolescents with type 1 diabetes mellitus. AB - Diabetes mellitus is characterized by microangiopathy and increased angiogenic response in various organs. Basic fibroblast growth factor (bFGF) as well as vascular endothelial growth factor (VEGF) are both angiogenic and are involved in vascular endothelial cell growth. The purpose of this study was to determine serum levels of bFGF and VEGF, in children and adolescents (youngsters) with type 1 diabetes mellitus, and correlate them with parameters reflecting the severity of the disease. Forty diabetic youngsters without clinical evidence of complications were compared with 30 healthy control subjects (mean age +/- SD, 14.3 +/- 3.6 and 13.8 +/- 3.6 y, respectively). Diabetes duration and metabolic control (expressed by glycosylated Hb) were (mean +/- SD) 6.2 +/- 3.8 y and 9.6 +/- 1.8%, respectively. bFGF and VEGF (pg/mL) were measured in serum samples by enzyme immunoassays, and both were not significantly different between the type 1 diabetes mellitus and the control group (p = 0.952 and p = 0.559, respectively). Restricting the analysis to the type 1 diabetes mellitus group, neither the duration nor the metabolic control of the disease showed any correlation with bFGF and VEGF serum levels, whereas a significantly positive correlation was found between the two examined angiogenic factors both in the diabetic (r = 0.3464, p = 0.025) and the control group (r = 0.4619, p = 0.0013). In conclusion, serum levels of bFGF and VEGF were not found to vary significantly in diabetic youngsters in relation to controls and had no correlation with the duration and metabolic control of the disease. Nevertheless, a positive correlation was found between these two angiogenic factors both in the type 1 diabetes mellitus and the control group. PMID- 9853921 TI - Antenatal glucocorticoids modulate the amplitude of pulsatile cortisol secretion in premature neonates. AB - We hypothesized that antenatal exposure to glucocorticoids influences subsequent pulsatile cortisol (F) secretion in premature neonates. To test this hypothesis, blood was sampled for plasma F determination via indwelling arterial lines at 15 min intervals for 6 h in 26 clinically stable neonates whose gestational ages were 25-33 wk. Deconvolution analysis was used to characterize F secretion and elimination. Pulsatile F secretion was observed in all neonates. Deconvolution estimates in eight neonates exposed to antenatal glucocorticoids (ANG group) were compared with those of 18 neonates not or only remotely exposed to ANG (No/RG group). The median amplitude of the F secretory burst of the ANG group was significantly less than that of the No/RG group [4.3 nmol/Lv x min and 9.2 nmol/Lv x min, respectively; p = 0.026 (Lv is liter of F distribution volume)]. The number and duration of F secretory bursts was similar for both groups: 5 bursts per 6 h, and 23 versus 16 min. By univariate linear regression analysis, mean arterial blood pressure correlated positively with F secretory burst frequency and F production rate (p = 0.0035, r = 0.55 and p = 0.0067, r = 0.52, respectively). We propose that ANG treatment modulates the amplitude of pulsatile F secretion in premature neonates. PMID- 9853922 TI - The relationship between stature, growth, and short-term changes in height and weight in normal prepubertal children. AB - Human growth is a nonlinear process with marked variation in growth rate during the short-term. It is not known how long-term height gain or stature is influenced by short-term changes in height and weight. This study has addressed these issues by using thrice weekly height and weight measurements during 1 year in 43 normal prepubertal children (aged 5.7-7.7 y) to construct individual height and weight velocity curves by regression analysis. The former were comprised of 3 to 6 growth spurts separated by stasis, whereas the latter were characterized by 2 to 5 periods of weight gain separated by periods of weight loss. Stepwise regression analysis to determine characteristics of these curves that influence stature and growth showed that height SD score was correlated to the mean absolute weight velocity amplitude (+), the mean length of height velocity peaks (-), and the number of periods of weight gain (-) (r2 = 38%). In contrast, change in height SD score (delta height SD score) was correlated to the number (+) and mean amplitude (+) of the periods of weight gain and the mean height velocity peak amplitude (+) (r2 = 44%). Examination of changes in height relative to weight during the year in the whole group revealed that height increased relative to weight in autumn and spring, whereas the reverse occurred during the winter months. We conclude that 1) both height and weight velocities during 1 year show a biphasic pattern, 2) there is seasonal variation in the short-term change in height relative to weight, and 3) prepubertal stature and the amount grown through the year are related to short-term changes in height and weight. Our data indicate that large but infrequent changes in weight with growth spurts of short duration are found in tall children. Good growth during the year was related to large but frequent gains in weight and large individual spurts in height. PMID- 9853923 TI - Adrenal function in preterm infants: ACTH may not be the sole regulator of the fetal zone. AB - The fetal zone of the adrenal gland is known to persist after preterm birth, but there is uncertainty as to how long adrenal fetal zone steroid production continues and how it is regulated. The purpose of this study was to test two hypotheses. First, that the urinary excretion of 3beta-OH-5-ene steroids persists until term, and then declines, as it does in full-term infants. Second, that the persistence of the fetal zone is due to continuing ACTH stimulation. A longitudinal observational study was undertaken in 22 preterm infants of 24-31-wk gestation. Sequential measurements were made of urinary 3beta-OH-5-ene steroids (fetal zone steroid metabolites), plasma dehydroepiandrosterone sulfate (DHEAS), and ACTH. Excretion of urinary 3beta-OH-5-ene steroids was 1500-2000 microg kg( 1) d(-1), persisting until term, and declining abruptly at approximately 42 wk postconceptional age (PCA), to levels comparable to term infants at the same PCA. Median plasma ACTH levels rose from <7.6 pg mL(-1) at 25-wk PCA to 34.5 pg mL(-1) at 46-wk PCA. Urinary 3beta-OH-5-ene steroids were highest when ACTH levels were lowest, and were declining when ACTH was rising. In four infants given dexamethasone, urinary excretion of 3beta-OH-5-ene steroids and plasma DHEAS were not suppressed fully, when plasma ACTH and cortisol, and urinary cortisol metabolites were. These data suggest that ACTH is not the sole regulator of the adrenal fetal zone steroid synthesis and that involution of the fetal zone is related to gestation rather than birth. PMID- 9853924 TI - Fluid abnormalities occur in the chronically cannulated mid-gestation but not late gestation ovine fetus. AB - Hydrops fetalis, with or without oligo- or polyhydramnios, is associated with very high fetal mortality. In many cases the causes are unknown. Chronically cannulated ovine fetuses have been used as animal models to study the regulation of fetal fluid balance. This study reports that the mid-gestation ovine fetus (70 +/- 1 d of gestation; term = 145-150 d) is susceptible to the development of fetal abnormalities (excess allantoic fluid--hydrallantois, with or without hydrops and hydranencephaly), when blood vessels in the neck are cannulated. Cannulation of one carotid artery and one jugular vein, or cannulation of a single jugular vein resulted in 5 out of 12 fetuses having abnormalities 1 wk later. In contrast, six fetuses at 115 d of gestation that had both carotids and one jugular vein ligated cranially and cannulated, developed hydranencephaly but no hydrops or hydrallantois. In the mid-gestation fetus hydrallantois [760 +/- 140 mL (n = 5) versus 104 +/- 23 mL (n = 7 controls), p < 0.001] occurred without alterations in the plasma concentrations of ACTH, cortisol, atrial natriuretic peptide, or aldosterone, as well as without anemia. Although the causes of the fluid abnormalities were not resolved, it is important to note the developmental differences in vulnerability. PMID- 9853925 TI - Effect of increasing dietary threonine intakes on amino acid metabolism of the central nervous system and peripheral tissues in growing rats. AB - The threonine content of most of the infant formulas currently on the market is approximately 20% higher than the threonine concentration in human milk. Due to this high threonine content the plasma threonine concentrations are up to twice as high in premature infants fed these formulas than in infants fed human milk. To study the effect of different threonine intakes on plasma and tissue amino acid concentrations, 24 young male Wistar rats were fed three experimental diets based on a mixture of bovine proteins with a whey protein/casein ratio of 60/40 with different threonine contents [group A, 0.86 g of threonine/100 g (n = 8); group B, 1.03 g of threonine/100 g (n = 8); group C, 2.21 g of threonine/100 g (n = 8)]. Eight animals were fed a typical rat diet based on bovine casein as controls. After a feeding period of 15 d, amino acids were measured in plasma and in homogenates of the cerebral cortex, brain stem, liver, and muscle. There was a significant correlation between threonine intake and plasma threonine levels (r = 0.687, p < 0.001). The plasma threonine concentration correlated significantly with the threonine concentration in the cortex (r = 0.821, p < 0.01) and the brain stem (r = 0.882, p < 0.01). There was a positive significant correlation between threonine and glycine concentrations in the cortex (r = 0.673, p < 0.01), and the brain stem (r = 0.575, p < 0.01), whereas the glycine concentration decreased with increasing threonine intakes in the liver and muscle. The presented data indicate that increasing the threonine in plasma leads to increasing brain glycine and thereby affects the neurotransmitter balance in the brain. This may have consequences for brain development during early postnatal life. Therefore, excessive threonine intake during infant feeding should be avoided. PMID- 9853926 TI - Phenylalanine and tyrosine metabolism in neonates receiving parenteral nutrition differing in pattern of amino acids. AB - Tyrosine is considered to be an indispensable dietary amino acid in the neonate, yet achieving adequate parenteral tyrosine intake is difficult due to its poor solubility. Increasing the supply of phenylalanine is the most common means of compensating for low tyrosine levels. Unfortunately, plasma phenylalanine concentrations are sometimes elevated in infants receiving high phenylalanine intake. This led us to study the phenylalanine and tyrosine metabolism in 16 neonates randomized to receive total parenteral nutrition with either a high or a moderate phenylalanine-containing amino acid solution. A primed, 24-h continuous stable isotope infusion of L-[1-13C]phenylalanine and L-[3,3-2H2]tyrosine was given to enable the measurement of phenylalanine and tyrosine kinetics. Results demonstrated that 1) phenylalanine hydroxylation was significantly greater in infants receiving high phenylalanine, 2) phenylalanine oxidation and percent dose oxidized was also significantly greater in infants receiving high phenylalanine, 3) apparent phenylalanine retention was greater in neonates receiving high phenylalanine, and 4) alternate catabolites of phenylalanine and tyrosine metabolism were significantly greater in infants receiving high phenylalanine compared with moderate phenylalanine. We conclude that neonates respond to increased parenteral phenylalanine intake by increasing their hydroxylation and oxidation rates. The greater oxidation of phenylalanine in infants receiving high phenylalanine in conjunction with the urinary excretion of alternate catabolites of phenylalanine and tyrosine suggests that the high phenylalanine intake may be in excess of needs. However, the lower apparent phenylalanine retention observed in infants receiving moderate phenylalanine suggests that the total aromatic amino acid level of moderate phenylalanine may be deficient for neonatal needs. PMID- 9853927 TI - Polysialylated neural cell adhesion molecule serum levels in normal children. AB - Serum concentrations of polysialylated neural cell adhesion molecule (PSA-NCAM), a developmentally regulated form of the NCAM, have been recently described to be elevated in children with rhabdomyosarcoma and neuroblastoma, proving PSA-NCAM to be a tumor marker of diagnostic relevance to these malignancies. The present investigation was undertaken to define age-dependent reference intervals in normal children. Serum concentrations of polysialylated NCAM were determined in 366 children aged newborn to 17 y and in 18 adult patients by an immunoluminescence assay using the polysialic acid-specific MAb 735. Serum levels in newborn children were 51.7 kU/L (mean +/- 12.0 kU/L SD), whereas in adult patients they were 9.9 kU/L (mean +/- 3.5 kU/l SD). Assigning the patients to 14 different age groups, a gradual decay of PSA-NCAM serum concentrations was observed, and therefore, mean levels and empirical interpolated percentiles were determined for every age group. Applying specially fitted logistic functions, two different sigmoid graphs were obtained describing the age-dependent decrease of serum PSA-NCAM during the neonatal period and during childhood. The age at which the levels reach half the initial value was located at 3.1 d (mean +/- 2 d SE) and 14 y (mean +/- 1 y SE), respectively. There was no difference between male and female individuals. Repeated measurements revealed variations below 10%. For the first time, our study describes serum levels of PSA-NCAM in children of different age and their gradual decay until adulthood. PMID- 9853928 TI - Genotype-phenotype association in infants with cystic fibrosis at the time of diagnosis. AB - The relationship between the most common disease-causing mutations, the clinical manifestation, and lung function was prospectively assessed in 60 infants (33 females, 27 males) with cystic fibrosis at time of diagnosis (age: 7.2 months; range: 0.8-23.8 months). Lung function was assessed by infants whole-body plethysmography. Age at time of diagnosis was independent from the genotype. Weight gain from birth until the time of diagnosis expressed in percent predicted of a normal population was lower in the 3905insT group (57.9 +/- 19.0%) compared with deltaF508 homozygotes (62.5 +/- 20.6%; n.s.) and the R553X group (85.9 +/- 10.9%; p < 0.005). Differences regarding lung function within the genetic groups are mainly related to pulmonary hyperinflation, measured by thoracic gas volume (TGV), present in 8 of 9 infants with 3905insT, differentiating this frameshift mutation (TGV of 7.0 +/- 3.6 SD-S) from the R553X mutation (TGV 2.1 +/- 4.6 SD-S; p < 0.02). It is concluded that the variable disease findings in infants with cystic fibrosis is clinically and functionally reflected by features already present at time of diagnosis. The degree of pulmonary hyperinflation is, at least partly, influenced by the genotype. PMID- 9853929 TI - Identification of a hereditary pancreatitis mutation in four West Virginia families. AB - Hereditary pancreatitis (HP) is the second most common cause of chronic childhood pancreatitis in the United States. Mutations in the cationic trypsinogen gene on chromosome 7 are known to cause HP. We identified four families in West Virginia with symptoms consistent with HP. To determine whether members of these families had defects in the trypsinogen gene, we tested for linkage between the HP gene and simple tandem repeat markers on chromosome 7q and screened for a specific mutation in the cationic trypsinogen gene. Two-point linkage analysis indicated that the disease gene is closely linked to three 7q markers (D7S661, D7S2511, and D7S1805). Restriction fragment length polymorphism analysis showed that all clinically affected members and nonpenetrant carriers from the four families carried a G to A mutation in the third exon of the trypsinogen gene. These findings indicate that this mutation is the cause of HP in the families in our study. The observation that most individuals who carry the mutation have symptoms of HP is consistent with the high but incomplete penetrance of the trait. The presence of a single mutation and a common linked haplotype indicates that the defective allele arose in an ancestor common to all four families. PMID- 9853930 TI - Sensitization to cow's milk proteins during refeeding of guinea pigs recovering from polydeficient malnutrition. AB - We have previously shown that milk sensitization aggravates intestinal dysfunction in the malnourished guinea pigs, suggesting that it may also impair the recovery from malnutrition. To test this hypothesis, the growing guinea pigs were malnourished by feeding only maize for 7 d and then were refed for 21 d with a balanced diet containing either intact or hydrolyzed cow's milk proteins. The control animals received the hydrolyzed milk protein diet for 28 d. After an initial period of total inhibition of growth owing to maize, guinea pigs gained weight regularly, with both balanced diets, and there was no evidence of mucosal damage at the end of the refeeding period. However, refeeding with intact milk proteins induced milk sensitization, which was demonstrated on the systemic level by the presence of anti-beta-lactoglobulin IgG1 antibodies, and on the local level by the intestinal anaphylaxis measured by the increase in short circuit current induced by beta-lactoglobulin (16.4 +/- 2.6 microA/cm2) in jejunal segments mounted in Ussing chambers. Such an immune sensitization was associated with impaired intestinal permeability, as both the ionic conductance (21.0 +/- 1.6 versus 14.6 +/- 0.7 mS/cm2) and the transepithelial fluxes of horseradish peroxidase (537 +/- 203 versus 152 +/- 28 ng/h x cm2) were significantly increased in guinea pigs refed with the intact milk proteins compared with controls. In contrast, there was no difference in intestinal permeability between controls and guinea pigs refed with the hydrolyzed milk protein diet. These data show that sensitization to cow's milk proteins can develop in guinea pigs recovering from severe malnutrition and may impair full intestinal repair. PMID- 9853931 TI - Adolescents with alpha1-antitrypsin deficiency have high alpha2-macroglobulin and low neutrophil lipocalin and elastase levels in plasma. AB - Eighteen-year-old adolescents with alpha1-antitrypsin (alpha1AT) deficiency have mostly normal lung function tests. We hypothesized that compensatory increases in other protease inhibitors and/or a decreased leukocyte activity might favorably affect the protease/protease-inhibitor balance in alpha1AT-deficient adolescents. At the age of 18 y 46 PiZZ (severe deficiency), 22 PiSZ (moderate deficiency), and 41 control subjects were studied. The plasma protease inhibitors alpha2 macroglobulin (alpha2M), alpha1-antichymotrypsin (Achy), and secretory leukocyte protease inhibitor (SLPI) were studied, and the protease elastase complexed with alpha1AT (HEAT) and neutrophil gelatinase-associated lipocalin (NGAL) as indicators of neutrophil leukocyte activity. Significantly higher concentrations of alpha2M were found in PiZ (p < 0.0001) and PiSZ (p < 0.0001) individuals compared with control subjects. The PiZZ and SZ adolescents had low levels of NGAL (p < 0.0001). Low levels of HEAT were found in PiZZ subjects (p < 0.0005). Higher concentrations of Achy were found in PiZZ (p < 0.04) and PiSZ (p < 0.05) individuals. Increased concentrations of alpha2M and Achy combined with decreased levels of HEAT and NGAL, indicating decreased leukocyte activity may, to some extent, compensate for the protease/protease inhibitor imbalance in the alpha1AT deficiency state. PMID- 9853932 TI - Neonatal neutrophil activation is a function of labor length in preterm infants. AB - To understand better the development of the neonatal immune system, we evaluated the role of labor length, gestational age, and mode of delivery on the expression of the neonatal neutrophil cell surface antigens CD11b, CD11c, CD15, CD33, and CD66b in premature newborns. Peripheral blood samples from 68 apparently healthy preterm infants were obtained within 12 h of birth and incubated with MAb to the CD antigens. Samples were lysed, fixed, and analyzed by flow cytometry. Multivariate analysis was used to study the simultaneous effect of the labor length and gestational age on the neonatal neutrophil cell surface antigen expression. A positive correlation was demonstrated between neutrophil antigen expression and labor length (p < 0.001-0.026) but not with the mode of delivery (p = 0.191-0.638). There was no significant correlation between expression of neutrophil antigens and gestational age at delivery (p = 0.057-0.866), except for CD15 (p = 0.010). Our results indicate labor length is a significant factor in neonatal neutrophil activation at birth. These findings are independent of gestational age in preterm newborns. Mode of delivery does not seem to influence neonatal neutrophil activation. The neutrophils of premature infants can be activated antenatally and/or during labor. PMID- 9853933 TI - Passive deformability of mature, immature, and active neutrophils in healthy and septicemic neonates. AB - Obstruction of narrow vessels by rigid neutrophils may contribute to ischemic organ injury. In septicemia, a substantial portion of the neutrophils may become activated and the number of circulating immature neutrophils may rise sharply. Volume and deformability of mature (PMN) and immature neutrophils in healthy preterm and full-term infants and in septicemic neonates were studied by means of a micropipette system. Membrane cytoplasm tongues were aspirated into 2.5-microm (diameter) pipettes over a period of 60 s. Volume and tongue growth of mature resting PMN were similar in healthy preterm and full-term neonates and adults. Compared with mature PMN (about 360 fl), the volumes of band cells (415 fl), metamyelocytes (470 fl), and less mature cells (myeloblasts, promyelocytes, and myelocytes; 490 fl) were significantly increased (p < 0.005). Final tongue lengths of band cells, metamyelocytes, and less mature cells were decreased by about 50, 60, and 70%, respectively, when compared with passive mature cells. In septic neonates, the percentage of immature neutrophils was increased, but the deformability and volume of the cell subpopulations were not affected by septicemia. Active PMN were characterized by pseudopod formation. More active PMN were found in group B streptococcal (14% of total PMN), gram-negative (12%), and Staphylococcus epidermidis septicemia (8%) than in healthy neonates and adults (4%). The main bodies of active PMN were less deformable than passive PMN, and the pseudopods showed very little membrane deformation. The increased number of rigid active and immature neutrophils may contribute to impaired microcirculation and the high risk for organ injury in septic patients. PMID- 9853934 TI - The cerebral hemodynamic response to asphyxia and hypoxia in the near-term fetal sheep as measured by near infrared spectroscopy. AB - This study examined the hypothesis that the cerebrovascular response to asphyxia of the late gestation sheep fetus is characterized by an increase in cerebrovascular resistance and a fall in cerebral blood flow (CBF) rather than the fall in resistance and increase in CBF which occurs in acute hypoxemia. In eight unanesthetized late gestation fetal sheep (123- to 125-d gestation) we evaluated continuous changes in carotid blood flow (CaBF) as an index of global CBF and total cerebral Hb concentrations as an index of global cerebral blood volume (CBV) using ultrasound flow probes and near infrared spectroscopy respectively. Asphyxia was induced by rapid and complete occlusion of the umbilical cord for 10 min. We also examined the fetal response to 1 h of acute 9% isocapnic hypoxia for comparison purposes. During hypoxia we observed a sustained increase in CaBF (p < 0.05) and CBV (p < 0.01) and a fall in carotid vascular resistance (p < 0.05). During asphyxia there was no significant rise in CBV, a fall in CaBF (p < 0.05), and a rise in carotid vascular resistance (p < 0.01). CaBF fell at a time when mean arterial pressure was elevated (p < 0.01). These data strongly suggest that fetal CBF does not increase and may even fall during severe asphyxia of rapid onset. Furthermore, our near infrared spectroscopy data show that the relative changes in total cerebral Hb concentrations may reflect the type and severity of the insult to which the fetus is exposed. PMID- 9853935 TI - American Pediatric Society Presidential Address 1998: what is the future for academic pediatrics? PMID- 9853936 TI - Society for Pediatric Research Presidential Address 1998: the SPR and 1-800-NO CLOTS: a common vision. PMID- 9853937 TI - Impact of the sequential IPV/OPV schedule on vaccination coverage levels--United States, 1997. AB - In January 1997, the Advisory Committee on Immunization Practices (ACIP) recommended adoption of a sequential inactivated poliovirus vaccine (IPV)-oral poliovirus vaccine (OPV) vaccination schedule. The schedule of injections of IPV at 2 months and 4 months of age, followed by OPV at 12-18 months and again at 4-6 years was intended to minimize the risk for vaccine-associated paralytic poliomyelitis (VAPP) while maintaining population immunity to the potential introduction of wild-type poliovirus. To determine whether this change may result in reduced or delayed vaccination coverage because parents or physicians might be reluctant to administer multiple injections at a single visit, CDC investigated the impact of the change to a sequential IPV-OPV vaccination schedule at two large West coast health maintenance organizations (HMOs). This report summarizes the results of the investigation and indicates that changing to an initial two doses of IPV was not associated with decreases in vaccination coverage levels of routinely recommended vaccinations. PMID- 9853938 TI - Fatal car trunk entrapment involving children--United States, 1987-1998. AB - During July-August 1998, at least 11 U.S. children died in three separate incidents of car trunk entrapment. This report summarizes these three incidents, describes characteristics of car trunk entrapment incidents involving children since 1987, and reviews measures to prevent children from becoming trapped in car trunks. The findings indicate that at least nine incidents of fatal car trunk entrapment involving children occurred during 1987-1998, that all incidents occurred in hot weather and involved children aged < or =6 years, and that these deaths were preventable. PMID- 9853939 TI - Forecasted state-specific estimates of self-reported asthma prevalence--United States, 1998. AB - Asthma is a chronic inflammatory disorder of the lungs characterized by episodic and reversible symptoms of airflow obstruction. During 1993-1994, an estimated 13.7 million persons in the United States reported having asthma, and from 1980 to 1994 the prevalence of self-reported asthma in the United States increased 75%. Despite this increase, surveillance data are limited for asthma at the state and local levels. To estimate the 1998 prevalence rate of asthma for each state, CDC analyzed national self-reported asthma prevalence data from 1995. This report summarizes the results of the analyses, which project that approximately 17 million persons in the United States have asthma. PMID- 9853940 TI - Abortion surveillance: preliminary analysis--United States, 1996. AB - For 1996, CDC compiled data about legal induced abortions from the 50 states, New York City, and the District of Columbia. The total number of legal induced abortions was available from all reporting areas; however, not all areas collected information about the characteristics of women who obtained abortions. This report presents preliminary data for 1996; final abortion data for 1996 will be published during spring 1999. PMID- 9853941 TI - Free combined anterolateral thigh flap and vascularized fibula for wide, through and-through oromandibular defects. AB - Through-and-through oromandibular defects require the greatest amount of soft tissue volume among the transfers for head and neck defects. A new method, a large anterolateral thigh flap combined with a vascularized fibula graft in a chimera fashion, has been used for two patients with wide through-and-through oromandibular defects. Among the candidates for such a large skin flap, the anterolateral thigh flap seems to be the best, for the following reasons. (1) Its pedicle, the lateral circumflex femoral system, has several major branches of equal size of anastomosis of the peroneal vessels. (2) As the majority of such patients with multiple previous surgery have lost recipient vessels near the mandible, the longest vascular pedicle is required. (3) There is no need for positional changes, and simultaneous flap elevation with the tumor resectioning is possible. (4) Use of the fibula allows for reconstruction of the entire mandible, if necessary. (5) Some of the shortcomings of individual donor sites for massive composite osteocutaneous flaps are minimized, because each component consists of two donor sites. (6) Operating time for this flap elevation is minimized, compared to that for massive composite osteocutaneous flaps, because the individual components can be obtained simultaneously by two teams. PMID- 9853942 TI - Pharyngoesophageal reconstruction with a tubed free radial forearm flap. AB - Various attempts at reconstruction of pharyngoesophageal defects after ablative surgery have been made to restore the function of the pharyngoesophagus. A tubed free radial forearm flap was used to reconstruct the pharyngoesophagus in 23 patients after resection of neoplasms from May 1989 to October 1995. Nineteen were males and four were females, the average patient age was 62.2 years. The follow-up ranged from 10 to 64 months (mean: 18 months). Oral intake within 3 weeks was possible in 18 patients (78 percent) The immediate postoperative complications were hematoma (n = 1), bleeding (n = 2), infection (n = 3), fistula (n = 4), and venous thrombosis (n = 1). A late complication was stricture of the lower anastomosing site (n = 3). The tubed free radial forearm flap has advantages over free jejunal transfer, including the larger caliber of the vascular pedicle, longer possible ischemic time, no laparotomy with less morbidity of the donor site, and better toleration of radiotherapy. Troublesome disadvantages include stricture and fistula formation at the suture sites. The authors modified the conventional free radial forearm flap to reduce complications. A small monitoring flap supplied by the septocutaneous branch of the radial artery was elevated to check the survival of the flap. During tubing, the vertical suture line was overlapped with a deepithelialized skin flap, and double layer sutures were done to prevent fistula. Two small triangular flaps were designed and inserted at the distal anastomotic site to prevent circular contracture. The outer-layer sutures were anchored to the surrounding rigid structure to withstand shrinkage and circular contraction. With this modification, the incidence of stricture and fistula formation was reduced to 13.0 percent and 17.4 percent, respectively, and these complications could be treated conservatively. PMID- 9853943 TI - Sensate myocutaneous latissimus dorsi flap. AB - A case using a sensate myocutaneous latissimus dorsi flap for oral defect reconstruction is reported. Preparation of the cutaneous branches of a latissimus dorsi autologous transfer and their subsequent anastomoses with sensitive nerves at the recipient site resulted in superficial and deep sensibility of the transfer, following a short rehabilitation period. As early as 4 months postoperatively, pain and pressure sensations could be evoked in the flap. Sharp and blunt discrimination and sense of vibration were found 6 months postoperatively. Nine months postoperatively, caloric differentiation could be elicited. PMID- 9853944 TI - Rotation plasty for osteosarcoma of the femur. AB - A 20-year-old female with right femoral osteosarcoma noted severe pain and swelling of the right thigh, and was transferred to the authors' hospital after suffering a pathologic fracture of the right femur. Plain x-ray disclosed a tumor shadow extending from the pathologic fracture of the distal portion of the femur to the center of the diaphysis. MRIs showed a broad lesion and hematoma, and invasion to the femoral vascular bundle was suspected. Angiograms revealed vascularization coincidental to the tumor, and an irregular vascular wall of the femoral artery, which was shifted posteriorly. After consultation with the patient and her family, rotation plasty was performed. The sciatic nerve was isolated and preserved; major vessels were sectioned proximally and distally because of tumor tissue invasion to the femoral artery and vein. After wide excision of the tumor, the leg was rotated outward 180 degrees, and osteosynthesis was performed between the remaining femur and the tibia, followed by microsurgical anastomosis of vessels. Five years postoperatively, no local recurrence or metastasis has been observed. The patient has excellent functional recovery without pain, and no other complications. PMID- 9853945 TI - Bilateral radial forearm free flaps for oral-cavity reconstruction. AB - Simultaneous primary tumors (tumors found at the same time or within 1 month of each other) are not uncommon in head and neck cancer. Most frequently, one lesion is small relative to the other; however, this is not always the case When two anatomically separate simultaneous primary tumors are large, a challenge for the reconstructive surgeon exists. The authors present a case of simultaneous primary tumors arising along the left and right mandibular gingiva. Following extirpation of the tumors, the defects were reconstructed with bilateral free radial forearm flaps. They conclude that this procedure is an excellent option for reconstruction of anatomically separate simultaneous defects of the head and neck region. This reconstruction provides an excellent tissue match for the intraoral region, and can be accomplished with minimal morbidity in a relatively short operative time. PMID- 9853946 TI - Reconstruction of intractable ulcer of the knee joint in Werner's syndrome with free latissimus dorsi myocutaneous flap. AB - The authors report the successful treatment of an intractable ulcer of the right knee in a patient with Werner's syndrome, using a free latissimus dorsi myocutaneous flap transfer. PMID- 9853947 TI - Effect of cryopreservation on survival of composite tissue grafts. AB - The problems in cryopreserving complex tissues remain poorly understood. In the present study, the viability of cryopreserved complex tissue, including skin and muscle flaps, and tail, was compared to simple artery and vein grafts in a rat model. Forty-four grafts, including the femoral artery (n = 15), femoral vein (n = 9), gracilis muscle flap (n = 8), groin cutaneous flap (n = 9), and tail (n = 3) were cryopreserved at -196 degrees C for 1 to 2 weeks, and then microvascularly transferred to analogous sites in inbred recipient animals. Three to 60 days post-transplant, all cryopreserved femoral arteries and veins were patent. Host groin cutaneous flaps with cryopreserved vessels grafted into the arterial pedicle survived with no complications However, severe endothelial-cell damage with extensive sloughing in the grafts was demonstrated at 30 min and 1 week following reperfusion, using factor VIII immunologic stain. Reendothelialization was evident at 2 months postoperatively. All cryopreserved composite tissue underwent early pedicle thrombosis The muscle and cutaneous tissue also showed gross and microscopic edema, with extensive red-cell extravasation within 30 min of reperfusion Cryopreservation thus can maintain the functional integrity of simple vessel grafts, but these grafts survive only as a conduit, given the severe endothelial-cell damage The failure of transferred complex cryopreserved tissue may reflect a combination of cellular or architectural damage to the capillary bed from ice formation during the freezing process and from reperfusion injury. PMID- 9853948 TI - Effect of distraction osteogenesis on the peripheral nerve: experimental study in the rat. AB - Distraction osteogenesis is the current method of choice for bone lengthening. Despite the gain in experience, various complications are reported, among them, adverse effects on peripheral-nerve function. In order to thoroughly investigate the effect of distraction on neural tissue, a distraction osteogenesis model in the rat was established, using the femur of 30 Sprague-Dawley rats. The animals were randomized in three groups, following different rates of distraction (0.5 mm, 1.0 mm, and 1.5 mm/day) for 50, 25, and 16 days, respectively, so that the final length of distraction was the same in all groups. The mean sciatic function index ranged near normal in all groups. All groups demonstrated decrease of conduction velocity and the area under the curve of the compound action potential, while morphologic alterations consisted of decrease in the number of axons and evidence of active degeneration. Animals in Groups 1 (0.5 mm/day) and 2 (1 mm/day) displayed comparable changes, while in Group 3 animals (1.5 mm/day), changes were significantly more adversely dramatic. The safest and fastest rate of distraction in this rat model was determined to be 1 mm/day. PMID- 9853949 TI - Effects of hematoma on the short-term fate of experimental microvenous autografts. AB - The early effects of hematoma on the healing pattern of interpositional vein grafts were studied in 60 rats. In all animals, vein grafts approximately 10-mm in length were used to bridge defects created in the femoral artery and vein on the same side, with a 1:1 graft-to-host diameter match. Grafted vessels were completely covered with hematoma and separated from the surrounding tissue in 30 animals, while the other 30 rats served as controls. Both groups were divided into six subgroups of five animals each, according to the age of the grafts: 1, 3, 5, 7, 14, and 21 days. Graft patency and healing were assessed both clinically and histologically 20/30 (66.6 percent) of the interarterial and 27/30 (90 percent) of the intervenous vein grafts in Group 1 (hematoma group), and 28/30 (93.3 percent) of the interarterial and 29/30 196.6 percent) of the intervenous vein grafts in Group 2 (controls) were found patent. The patency rate of the interarterial vein grafts in the hematoma group was statistically significantly lower than in the control group and than in the intervenous vein grafts of the same group (p<0.005). The majority of interarterial vein-graft failures occurred after postoperative day 7 in Group 1. Patent specimens were examined under scanning electron and light microscopy. The authors concluded that perivascular hematoma caused spasm and flow disturbance, prolonged vessel-wall ischemia, and severe vessel-wall injury in the arterial circulation of the microvenous grafts. It also delayed the healing process and subjected the grafts to the development of occlusive mural thrombus. In contrast, the healing pattern of the intervenous vein grafts was not markedly influenced by perivascular hematoma, and equilibrium between thrombogenic and antithrombogenic factors on the graft surface was restored over a short period of time. PMID- 9853950 TI - Immunomagnetic cell sorting--pushing the limits. PMID- 9853951 TI - Recombinant breast carcinoma-associated mucins expressed in a baculovirus system contain a tumor specific epitope. AB - Mucins are highly immunogenic glycoproteins that are abundantly expressed by breast and other adenocarcinomas. In order to progress in the understanding of the structure immunity relationship of the breast tumor associated mucin and normal tissue mucin, two forms of breast carcinoma associated mucin, muc7-BV and pem-BV, were expressed in a baculovirus expression system. The muc7-BV was constructed by inserting the seven tandem repeats of mucin core cDNA fragment into transfer vector pAc360, forming a fusion protein containing 14 amino acids of the baculovirus polyhedrin N-terminus. The pem-BV was constructed by cloning full-length mucin cDNA into the transfer vector pVL1392. The recombinant mucins were purified using immunoaffinity chromatography. The purified muc7-BV and pem BV had molecular weights of 28 and 59 kd, respectively. No carbohydrate was detected on these recombinant mucins and is speculated to explain why both forms of recombinant mucin showed strong affinity to tumor-specific monoclonal antibody SM3. These recombinant mucins may have the potential value to develop vaccines against breast and other adenocarcinomas and to induce cytotoxic T-lymphocyte lines for immunotherapy of the same. PMID- 9853952 TI - Engineering the isoelectric point of a renal cell carcinoma targeting antibody greatly enhances scFv solubility. AB - BACKGROUND: The murine A6H monoclonal antibody targets a cell surface antigen associated with renal cell carcinoma with high specificity and excellent biodistribution properties. Tumor to blood ratios of > 40:1 have been achieved in clinical studies. OBJECTIVES: In order to generate an antibody engineering system that would allow the construction of improved derivatives for diagnostics and therapeutics, a single-chain Fv antibody (scFv) derived from A6H was constructed. The initial single-chain Fv, constructed with a cysteine residue and hexa histidine sequence at the C-terminus, displayed a limited solubility of 100 microg/ml at pH 7.4. The low solubility and refolding yield of the original single-chain Fv required that a more soluble variant be designed and constructed. STUDY DESIGN: We hypothesized that lowering the pI of the scFv antibody away from the physiological range would yield a more soluble antibody. A derivative was thus subsequently engineered with five glutamic acid residues followed by the cysteine and hexa-histidine residues. The cysteine was included to provide a conjugation site for future radiolabeling studies. RESULTS: The redesigned A6H single-chain Fv has a predicted pI of 6.1, relative to 7.5 for the native scFv. The redesigned A6H scFv displayed a greatly enhanced solubility of > 15 mg/ml at pH 7.4. Both the original scFv and the redesigned single-chain Fv exhibited a strong tendency to form dimers and soluble high molecular weight aggregates. The monomer and disulfide bonded dimer were separated from the aggregates and complete cell binding isotherms were obtained, demonstrating that the purified A6H scFv retains much of the activity of the parent monoclonal. CONCLUSION: The addition of glutamic acid to the C-terminus of poorly soluble scFv antibodies could provide a straightforward avenue for improving their solubility properties. The increased solubility of the A6H scFv allowed the purification of the monomeric and dimeric species from the soluble aggregated species. PMID- 9853953 TI - Deposition of genetically engineered human antibodies into the egg yolk of hens. AB - To determine if human immunoglobulins (hIg) are capable of being transported into the hen's egg, 10 microg each of purified hIgG and hIgA were intravenously injected into SC Hyline(TM) hens and their presence in egg yolk and egg white was determined by ELISA. In both cases deposition into the egg yolk was observed and in the case of hIgA, deposition was also observed in the egg white. Two stably transfected DT40 cell lines secreting recombinant human IgG3 and IgA (rhIgG3 and rhIgA) were injected into laying hens. The DT40 cells colonized the host and rhIgG3 and rhIgA were deposited in egg yolk. Deposition of rhIgA was also observed in the egg white. These data demonstrate that human immunoglobulins and other foreign proteins may be targeted to the chicken's egg. In view of the high rate of reproduction, the short generation interval, the high rates of egg production and the extensive infrastructure to fractionate egg yolk proteins, it should be possible to produce large amounts of foreign protein in the eggs of transgenic chickens. PMID- 9853955 TI - IgM secretory tailpiece drives multimerisation of bivalent scFv fragments in eukaryotic cells. AB - BACKGROUND: The monoclonal antibody (mAb) TP-3 binds selectively to human and canine osteosarcoma (OS) cells and is therefore a potential candidate for use as a targeting agent in radioimmunoimaging and therapy of OS metastases. However, intact murine mAbs have several drawbacks such as large size, delayed blood clearance and high immunogenicity, all of which can be overcome by genetic engineering. OBJECTIVES: To construct and express bivalent and multivalent TP-3 scFv fragments from the mammalian expression vector, pLNO. This vector has unique restriction sites for simple cassette cloning of any individual variable (V) and constant (C) genes and has previously been used for expression of intact chimeric TP-3 mAbs and Fab fragments. Furthermore, it is also suitable for expression of any modified V region, such as a scFv fragment, fused to any modified C region or to non-immunoglobulin protein sequences. STUDY DESIGN: Six different constructs were made; three scFv-CH3 fragments that differed in the design of linker between the scFv fragment and the IgG CH3 domain. These constructs were also made with the IgM secretory tailpiece (microtp) attached to the C terminus. RESULTS: All constructs were secreted as bivalent antibody fragments with a molecular weight of about 100 kDa. A band corresponding to a dimer appeared in all the supernatants from TP-3 scFv-CH3 producing cells, whether microtp was present or not, whereas higher orders of multimers were not seen. However, pulse chase analyses of the cells revealed that a small fraction of higher order polymers was formed from genes including the fragment encoding microtp and that microtp conferred retention both to monomers and intermediate polymers. The recombinant TP-3 antibody fragments were shown to bind human OS cells. CONCLUSION: Recombinant mAb fragments can be designed and cloned into the mammalian expression vector, pLNO. This vector is flexible in the sense that the genes encoding such fragments can be expressed from either cDNA or from genomic DNA. A microtp attached to the CH3 domain in these fragments was sufficient to drive polymerization, however inefficiently and intracellular retention of both monomers and intermediate polymers was observed. PMID- 9853954 TI - A novel tumor-specific human single-chain Fv selected from an active specific immunotherapy phage display library. AB - A colon tumor-associated antigen, CTAA 28A32-32K (CTA # 2E), related to the annexin family of proteins, was initially identified by its reactivity with a low affinity human IgM monoclonal antibody (mAb), 28A32. Both in vitro lymphoproliferative assays with human peripheral blood lymphocytes and delayed type hypersensitivity responses in patients immunized with autologous colon tumor cells indicated that CTA # 2E elicits potent T cell mediated responses and may be an important antigen in the development of a generic colorectal vaccine (Pomato et al. Vaccine Res 1994;3:145-161). A CTA # 2E-specific, murine hybridoma-derived mAb, 5-11A, which recognizes the amino-terminus of the tumor-associated antigen, exhibited qualitative human colon tumor-specific immunohistochemical reactivity. To rapidly develop a human mAb with similar antigen specificity and tumor reactivity as the murine 5-11A mAb, antibody phage display technology was employed. Two human antibody phage display libraries with 3.1 x 10(7) and 2.3 x 10(8) members were prepared from the variable region genes expressed by circulating B cells of patients undergoing active specific immunotherapy (ASI) with autologous tumor cells, predominantly from the colon, admixed with Bacille Calmette-Guerin (BCG). A CTA # 2E-reactive human single-chain (sc)Fv was selected by panning the larger library on decreasing concentrations of biotinylated tumor associated antigen in solution. It exhibited similar antigen specificity as the murine hybridoma-derived 5-11A scFv, requiring the presence of the CTA # 2E amino terminus for reactivity. This human scFv exhibited qualitative human colon tumor specific immunohistochemical reactivity when displayed as a gene III fusion protein on phage. When reconstructed and expressed as an intact human IgG1, K mAb, its qualitative colon tumor-specificity was unaltered. Two other CTA # 2E reactive human scFvs were selected from the smaller library by panning initially on decreasing concentrations of CTA # 2E coated to polystyrene and then on biotinylated CTA # 2E in solution. These human scFvs, which exhibited modest reactivity with different epitopes on the CTA # 2E antigen, did not exhibit human colon tumor-specific immunohistochemical reactivity. PMID- 9853956 TI - A system for stable indirect immobilization of multimeric recombinant proteins. AB - BACKGROUND: To perform an ELISA or for panning phage particles which display recombinant proteins, one of the reactants is immobilized on solid phase. Immobilization in ELISA is generally performed by passive adsorption of ligands to plastic. However, protein is denatured during the adsorption process. This may result in low efficiency interaction between ligands and receptors which depend on native structures. In contrast, indirect immobilization has been shown to prevent protein denaturation. OBJECTIVES: The aim was to develop a system that allows efficient and stable indirect immobilization of a variety of recombinant multimeric proteins to solid phase. RESULTS: A new vector was constructed which allows the expression of up to three proteins linked by the Jun/Fos leucine zipper. Purification of the resulting protein was achieved by Ni+ affinity chromatography utilizing the 6xHis-ABP (albumin binding protein) protein fused to the N-terminus of the Jun polypeptide. The high binding affinity of ABP to rat serum albumin (RSA) was exploited for indirect immobilization of recombinant proteins to solid phase. In an enzyme linked assay, the binding of ABP to immobilized RSA was shown to be 10-1000 times more efficient than other immobilization systems. Using the ZZ IgG binding domain of staphylococcal protein A as bait, the RSA-ABP immobilization system was successfully used to screen and enrich IgG Fc encoding DNA fragments from a cDNA phage library. CONCLUSION: The newly designed vector termed pJuFoexpress allows production and purification of multimeric protein complexes linked by the Jun/Fos leucine zipper. Without chemical modifications, the recombinant proteins can be immobilized indirectly to solid phase. The immobilization results in the stable display of native, biologically active proteins which can be used in ELISA and phage display systems. PMID- 9853957 TI - Migration of hypoglossal myoblast precursors. AB - The intrinsic hypoglossal musculature develops from precursor myoblasts which undergo long-range migration from the occipital somites to the tongue. Little detail is known about the precise spatiotemporal pathway taken by these cells or the factors controlling migration. In this study, chick/quail chimeras in which the occipital paraxial mesoderm is quail derived, reveal that the pathway taken by the tongue muscle progenitors is both complex and highly specific. Precursor myoblasts are Pax-3 positive cells which descend from the somite and migrate around the pharyngeal endoderm. They then course rostrally, following the base of the pharynx, remaining in a tight strand. We have examined a number of factors implicated in the control of migration of the hypoglossal precursors. Replacement of the occipital somites with those originating in the flank reveals that intrinsic differences do not exist between these somites with respect to their capacity to respond to migratory cues. The lack of high level HGF/SF expression along the pathway of the migrating hypoglossal precursors suggests that this factor is not involved in the actual process of migration of the hypoglossal precursors to the tongue. The pathway followed by the migrating precursors is identical to that of both the developing hypoglossal nerve and the circumpharyngeal crest--a subpopulation of the cranial neural crest, and importantly these populations utilize this pathway before the myoblast precursors. However, ablation neither of the hypoglossal nerve nor of the neural crest results in a perturbation in the ability of this Pax-3 positive population to migrate. This demonstrates that migration of the precursors is independent of both of these cell populations, and that it is controlled by the peripheral tissues. PMID- 9853958 TI - A role for neural cell adhesion molecule in the formation of the avian inner ear. AB - The inner ear forms by a series of folds within an ectodermal placode. Previous work has shown that changes in surrounding tissues play a more prominent role in invagination than changes in the cytoskeleton of the primordium. Interference with the integrity of the extracellular matrix causes abnormalities in the folding process, primarily related to abnormalities in the paraxial mesoderm which lies ventral to the placode. In this study, the role of the neural cell adhesion molecule (N-CAM) was investigated, based on the expression of this component of the plasmalemma at the time the otic placode begins to fold. Microinjection of blocking antibodies to N-CAM into the paraxial mesoderm adjacent to the otic placode resulted in two major classes of defects, detachment of the primordium from the neural tube and interference with formation of the folds. Microinjection of saline, control immunoglobulin, or antibody against cytoplasmic domain had no effect. These defects correlate with the pattern of N CAM expression at the time of injection, along the neural ectoderm and otic epithelium and the mesenchyme cells ventral to the primordium. It seems likely that N-CAM is playing a role in heterophilic associations rather than through the homophilic binding domain during formation of the otic vesicle. PMID- 9853959 TI - Ectopic expression of hoxb2 after retinoic acid treatment or mRNA injection: disruption of hindbrain and craniofacial morphogenesis in zebrafish embryos. AB - To investigate pattern formation in the vertebrate hindbrain, we isolated a full length hoxb2 cDNA clone from zebrafish. In a gene phylogeny, zebrafish hoxb2 clusters with human HOXB2, and it maps on linkage group 3 along with several other loci whose orthologues are syntenic with human HOXB2. In the hindbrain, hoxb2 is expressed at high levels in rhombomere 3 (r3), lower levels in r4, still lower in r5, and at undetectable levels in r6. In r7, r8, and the rostral spinal cord, hoxb2 is expressed at a lower level than in r5. Lateral cells appearing to emanate from r4 express both hoxb2 and dlx2, suggesting that they are neural crest. Overexpression of hoxb2 by mRNA injections into early cleavage stage embryos resulted in abnormal morphogenesis of the midbrain and rostral hindbrain, abnormal patterning in r4, fusion of cartilage elements arising from pharyngeal arches 1 and 2, and ectopic expression of krx20 and valentino (but not pax2, rtk1, or hoxb1) in the rostral hindbrain, midbrain, and, surprisingly, the eye. Treatments with retinoic acid produced a phenotype similar to that of ectopic hoxb2 expression, including ectopic krx20 (but not valentino) expression in the eye, and fusion of cartilages from pharyngeal arches 1 and 2. The results suggest that hoxb2 plays an important role in the patterning of hindbrain and pharyngeal arches in the zebrafish. PMID- 9853960 TI - Odontogenic epithelium induces similar molecular responses in chick and mouse mandibular mesenchyme. AB - Previous observations have shown that, during the initiation phase of odontogenesis, signals from mouse odontogenic epithelium can elicit teeth in non odontogenic but neural crest-derived mesenchyme isolated from ectopic sites including chick mandibular mesenchyme. In the present study the formation of ectopic tooth buds and dental mesenchyme in chick mandibular mesenchyme was examined using heterospecific recombinations between E11 mouse odontogenic epithelium and stage 23 chick lateral mandibular mesenchyme. Both morphological criteria and chick-specific probes for Msx-1, Msx-2, and Bmp-4 mRNAs were used as markers for early dental mesenchyme. Our results demonstrated that interactions of mouse odontogenic epithelium with chick mandibular mesenchyme induce early changes in the chick mandibular mesenchyme including the appearance of a translucent zone, cell proliferation, and induction of expression of Msx-1, Msx 2, and Bmp-4, which have been shown to be associated with the formation of dental mesenchyme. In addition, tooth bud-like structures that resemble E13 tooth buds in vivo both morphologically and in their patterns of gene expression formed after 6 days in the heterospecific recombinations. The tooth bud-like structures consist of invaginated mouse mandibular epithelium and condensed chick mandibular mesenchyme expressing high levels of Msx-1 and Bmp-4, but undetectable levels of Msx-2. Unlike the induction of Msx-1, Msx-2, and Bmp-4 in the underlying mesenchyme, which is specific for signals derived from odontogenic epithelium, the induction of a translucent zone and cellular proliferation in the underlying mesenchyme may be related to the growth-promoting potential of embryonic epithelia and not be specific to signals derived from the odontogenic epithelium. Similar to mouse odontogenic epithelium, agarose beads soaked in recombinant BMP 4 induced a translucent zone, cellular proliferation, and expression of Msx-1, Msx-2, and Bmp-4 in chick mandibular mesenchyme after 24 hours. These observations together showed that avian mandibular mesenchyme has odontogenic potential that is expressed upon interactions with inductive signals from mouse odontogenic epithelium. Similar to odontogenesis in vivo, formation of dental mesenchyme in chick mandibular mesenchyme is mediated by the activation of Msx-1, Msx-2, and Bmp-4. PMID- 9853961 TI - Bmp-2 downstream targets in mesenchymal development identified by subtractive cloning from recombinant mesenchymal progenitors (C3H10T1/2). AB - ABmp-dependent in vitro model was used to identify cDNAs during the manifestation of mesenchymal lineages. This model involves the recombinant expression of Bmps (Bmp-2, Bmp-4-7) in murine mesenchymal C3H10T1/2 progenitors, which leads to the differentiation into three lineages: the osteogenic, the chondrogenic and the adipogenic lineage, albeit in varying efficiencies. By subtractive cloning, 21 Bmp-2-regulated cDNAs from C3H10T1/2 mesenchymal progenitors were identified; 20 were related to known sequences and 1 was not. During mouse embryonic development, many of these cDNAs are expressed in chondrogenic, osteogenic, and in adipogenic tissues. Novel findings include a G0/G1 switch gene (G0S2), which was demonstrated to be predominantly expressed in adipose tissue during late murine embryonic development. Furthermore, the membrane-standing glycoprotein autotaxin (ATX) is expressed, at precartilage condensations, joint regions, and during tooth development. An as yet undescribed cDNA, 29A, which encodes a putative secreted factor, is expressed in developing osteo-/chondrogenic tissues of vertebrae, ribs, tooth, and the limb bud. C3H10T1/2-progenitors, therefore, may serve as a legitimate model for the investigation of the Bmp-mediated events during mesenchymal differentiation. PMID- 9853962 TI - Ectopic expression of tropomyosin promotes myofibrillogenesis in mutant axolotl hearts. AB - Expression of tropomyosin protein, an essential component of the thin filament, has been found to be drastically reduced in cardiac mutant hearts of the Mexican axolotl (Ambystoma mexicanum) with no formation of sarcomeric myofibrils. Therefore, this naturally occurring cardiac mutation is an appropriate model to examine the effects of delivering tropomyosin protein or tropomyosin cDNA into the deficient tissue. In this study, we describe the replacement of tropomyosin by using a cationic liposome transfection technique applied to whole hearts in vitro. When mouse alpha-tropomyosin cDNA under the control of a cardiac-specific alpha-myosin heavy chain promoter was transfected into the mutant hearts, tropomyosin expression was enhanced resulting in the formation of well-organized sarcomeric myofibrils. Transfection of a beta-tropomyosin construct under control of the same promoter did not result in enhanced organization of the myofibrils. Transfection of a beta-galactosidase reporter gene did not result in the formation of organized myofibrils or increased tropomyosin expression. These results demonstrate the importance of alpha-tropomyosin to the phenotype of this mutation and to normal myofibril formation. Moreover, we have shown that a crucial contractile protein can be ectopically expressed in cardiac muscle that is deficient in this protein, with the resulting formation of organized sarcomeres. PMID- 9853963 TI - Adenoviral-mediated expression of antisense RNA to fibroblast growth factors disrupts murine vascular development. AB - Fibroblast growth factors (FGFs) are expressed in the developing embryo and are postulated to regulate embryonic and vascular growth. The goal of this study was to elucidate the role of basic fibroblast growth factor (FGF-2) in early murine embryonic cardiovascular development in the mouse embryo. Gestation day 7.5 embryos were harvested and placed in culture, and 12 hr later replication defective adenovirus (0.5 x 10(6) plaque forming units) encoding either beta galactosidase or antisense FGF-2 RNA was injected into the sinus venosus of the cultured embryos. Embryos receiving only replication-defective adenovirus expressing the beta-galactosidase gene continued to develop normally over the next 12 hr. In contrast, those receiving adenovirus encoding antisense FGF-2 RNA displayed marked morphogenetic abnormalities, including cessation of growth and abnormal yolk sac vascular development, even though the embryonic hearts continued to beat. Abnormal development of the yolk sac vasculature was confirmed by microangiography and by histologic examination. Coinjection of virus carrying FGF-2 cDNA in the sense orientation reversed the effects of antisense FGF-2 RNA expression. These results confirm the efficacy of the replication-defective adenovirus for targeting gene expression to the developing vasculature and provide evidence for a critical role of FGF in the normal vascular assembly in the early embryo. Cessation of embryonic growth on expression of antisense FGF-2 RNA was most likely attributable to failure of efficient circulation of the early embryonic blood cells from the yolk sac into the embryo. PMID- 9853964 TI - Gpc3 expression correlates with the phenotype of the Simpson-Golabi-Behmel syndrome. AB - Interest in glypican-3 (GPC3), a member of the glypican-related integral membrane heparan sulfate proteoglycans (GRIPS) family, has increased with the finding that it is mutated in the Simpson-Golabi-Behmel overgrowth syndrome (Pilia et al. [1996] Nat. Genet. 12:241-247). The working model suggested that the membrane bound protein acts locally to limit tissue and organ growth and that it may function by interacting with insulin-like growth factor 2 (IGF2) to limit its local effective level. Here we have tested two predictions of the model. In situ hybridization with the mouse gene cDNA was used to study the expression pattern during embryonic and fetal development. In agreement with predictions, the gene is expressed in precisely the organs that overgrow in its absence; and the patterns of expression of Gpc3 and those reported for Igf2 are strictly correlated. PMID- 9853965 TI - sFRP-2 is a target of the Wnt-4 signaling pathway in the developing metanephric kidney. AB - Members of the Wnt family of secreted glycoproteins act as short-range signaling molecules in vertebrate embryogenesis. Previous work has shown that Wnt-4 is required for kidney development. Mice lacking functional Wnt-4 fail to form pretubular cell aggregates. Wnt-4 acts as an autoinducer of the mesenchymal to epithelial transition underlying nephron development. We have identified a member of the gene family encoding secreted frizzled related proteins (sFRP), putative Wnt antagonists, that shows overlapping expression with Wnt-4 in aggregating mesenchyme and simple epithelial bodies during metanephric development. sFRP-2 expression is absent in metanephric mesenchyme of kidneys mutant for Wnt-4 and is coinduced with Wnt-4 in isolated metanephric mesenchyme by cells expressing Wnt 4. The cysteine-rich domain of sFRP-2 binds to Wnt-4 as shown by coimmunoprecipitation experiments. Hence, sFRP-2 is a target of the Wnt-4 signaling pathway in the metanephric kidney and may modulate Wnt-4 signaling. sFRP-2 expression is highly dynamic and specific during other aspects of embryogenesis. sFRP-2 is expressed in subpopulations of ependymal cells in spinal cord and brain, in the developing eye, in limb bud mesenchyme, in the heart, and strongly in skeletogenic condensations of facial bones, suggesting widespread interaction with other members of the Wnt gene family during embryogenesis. PMID- 9853966 TI - Impaired growth and differentiation of diploid but not immortal melanoblasts from endothelin receptor B mutant (piebald) mice. AB - Endothelin 3 (Edn3) and its preferred receptor, endothelin receptor B (Ednrb), are implicated in development, especially that of two neural-crest-derived cell lineages: melanocytes and enteric ganglion cells. Mice and humans with a null mutation at either locus can show major deficiencies in both cell types: congenital white spotting and aganglionic megacolon (Hirschsprung disease in human). Numbers of early (migrating) embryonic melanoblasts are low in Ednrb(ls) mutant mice, while added Edn3 appears to promote the growth of melanocyte precursors in neural crest cultures. However, it is hard to assess cell differentiation in these mixed cultures, and it is not known whether Ednrb has any role in the postnatal melanocytic lineage. We have therefore studied primary cultures of neonatal melanoblasts homozygous for the piebald (Ednrb(s)) mutation. These mutant melanoblasts showed severe impairment of both net cell growth and differentiation compared to wild-type melanoblasts. They were also unresponsive to stimulation of growth by cholera toxin. We have established three immortal lines of melanoblasts and one of melanocytes homozygous for Ednrb(s). These immortal lines, however, had no detectable deficiency of growth or differentiation as judged by cell counts, induced pigmentation and immunocytochemistry for melanocytic markers. Consistent with this, neither Ednrb nor Edn3 mRNA was detected in 3/3 tested immortal lines of mouse melanoblasts and 5/5 lines of melanocytes, of various genotypes. We also report for the first time a method to grow immortal melanoblasts in pure culture, without feeder cells. PMID- 9853967 TI - Region-specific expression of chicken Sox2 in the developing gut and lung epithelium: regulation by epithelial-mesenchymal interactions. AB - In situ analysis of the chicken cSox2 gene, a member of the transcription factor family containing an Sry-like high-mobility group (HMG) box, demonstrated localized expression in the embryonic endoderm. Transcripts of cSox2 appeared before commencement of morphogenesis and cytodifferentiation in the rostral gut epithelium from the pharynx to the stomach. The caudal limit of cSox2 expression coincided with that of the region competent for proventricular differentiation and to the rostral limit of the domain of CdxA, a homologue of Drosophila caudal. During morphogenesis, the level of transcripts of cSox2 decreased in epithelia invaginating into surrounding mesenchyme to form glandular or tubular structures, such as the primordia of the thyroid and lung, glandular epithelium of the proventriculus, and secondary bronchus of the lung. Tissue recombination experiments demonstrated that cSox2 expression is regulated by the underlying mesenchyme as well as morphogenesis and cytodifferentiation. The results suggest that cSox2 plays pivotal roles in generating morphologically and physiologically distinct types of epithelial cells in the gut. PMID- 9853968 TI - Specification and migration of melanoblasts at the vagal level and in hyperpigmented Silkie chickens. AB - The final pattern of neural crest derivatives used to be believed to be the result of unspecified neural crest cells haphazardly entering migratory paths and then receiving cues unique to that path that direct their differentiation. An alternative model, which we have coined the phenotype-directed model, is that neural crest cells are fate-specified first and then select a migratory pathway based on their developmental specification. Support for this model comes from recent studies demonstrating that, at the thoracic level, neural crest cells are specified as melanocyte precursors (melanoblasts) prior to entering the dorsolateral path, and that only melanoblasts have the ability to migrate dorsolaterally. Here we examine two examples of melanocyte patterning in birds that apparently contradict this model. The first is neural crest at the vagal level, where early crest cells migrate dorsolaterally and enter the branchial arches. Despite the fact that these cells migrate dorsolaterally (suggesting that they are melanoblasts), branchial arch-derived neural crest cells fail to differentiate as melanocytes in vitro. These observations suggest that the branchial arch environment may not support the survival or differentiation of melanogenic neural crest cells. The second example is the hyperpigmented Silkie chickens, which exhibit extensive internal pigmentation. The Silkie defect has been linked to a difference in the neural crest migratory environment that potentially causes (or allows) unspecified neural crest cells to undergo melanogenesis in the ventral path. In both of these situations, it appears that the final distribution of pigment cells is controlled by environmental factors, which would contradict the phenotype-directed model. Here we show that the final pattern of melanocytes at the vagal level and in Silkie chickens reflects the migratory behavior of lineage-specified melanoblasts, as predicted by the phenotype-directed model. At the vagal level, the early, dorsolaterally migrating crest cells that colonize the branchial arches are not melanoblasts and are biased against melanogenesis in vitro. Melanoblasts are not specified until later, just prior to a second wave of dorsolateral migration, and although these cells migrate dorsolaterally they do not invade the branchial arches. In Silkie embryos, melanoblasts are specified late and only invade the dorsolateral path after they have been specified. Unlike quail and White leghorn melanoblasts, however, Silkie melanoblasts also migrate ventrally, but again only after they are specified. PMID- 9853969 TI - Eya1 expression in the developing ear and kidney: towards the understanding of the pathogenesis of Branchio-Oto-Renal (BOR) syndrome. AB - Branchio-Oto-Renal (BOR) syndrome is an autosomal dominant, early developmental defect characterised by varying combinations of branchial (fistulas, sinuses, and cysts), outer, middle and inner ear, and renal anomalies. The gene underlying this syndrome, EYA1, is homologous to the Drosophila developmental gene eyes absent which encodes a transcriptional co-activator required for eye specification. We report here the temporal and spatial pattern of expression of the murine homologue, Eya1, throughout ear and kidney development in relation to the anomalies of BOR syndrome. The expression of Eya1 in the branchial arch apparatus (namely in the 2nd, 3rd, and 4th branchial clefts and pharyngeal pouches) at embryonic day (E)10.5, can be correlated with the branchial fistulas, sinuses, and cysts but not with the outer and middle ear anomalies. In contrast, Eya1 is expressed during the slightly more advanced stage of outer and middle ear morphogenesis at E13.5, in the mesenchyme adjacent to the first branchial cleft (the cleft will give rise to the external auditory canal and the surrounding mesenchyme to the auricular hillocks) and surrounding the primordia of the middle ear ossicles, and in the epithelium of the tubotympanic recess (the future tympanic cavity). During early inner ear development, Eya1 is expressed in the ventromedial wall of the otic vesicle (the site of the future sensory epithelia), in the statoacoustic ganglion, and in the periotic mesenchyme, consistent with the cochlear anomalies and sensorineural hearing loss of BOR syndrome. Subsequently, Eya1 expression is observed in the differentiating hair and supporting cells of the sensory epithelia, as well as in the associated ganglia, and persists after differentiation has taken place. This suggests that, in addition to a role in the morphogenetic process, Eya1 could also be implicated in the differentiation and/or survival of these inner ear cell populations. Finally, Eya1 expression in the condensing mesenchymal cells of the kidney is consistent with the excretory and collecting system anomalies of BOR syndrome. From the comparison of the Eya1 and Pax2 expression patterns during ear and kidney development, a contribution of these two genes to the same regulatory pathway can only be suggested in the mesenchymal-epithelial transition directing renal tubule formation. PMID- 9853970 TI - PDGF-A and PDGFR-alpha regulate tooth formation via autocrine mechanism during mandibular morphogenesis in vitro. AB - Platelet-derived growth factor A (PDGF-A) binding to the PDGF receptor alpha (PDGFR-alpha) mediates signal transduction processes related to DNA synthesis, cell migrations, cytodifferentiation, and wound healing. Recent studies indicate that PDGFR-alpha functions during cranial neural crest cell migrations and first branchial arch morphogenesis (Stephenson et al. [1991] Proc. Natl. Acad. Sci. USA 88:6-10; Morrison-Graham et al. [1992] Development 115:133-142; Hu et al. [1995] Int. J. Dev. Biol. 39:939-945; Soriano [1997] Development 124:2691-2700). The present studies were designed to test the hypothesis that PDGF-A, interacts with its cognate receptor PDGFR-alpha via an autocrine mechanism that regulates the timing, rates, and size of embryonic mouse tooth morphogenesis. Both PDGF-A and PDGFR-alpha transcripts were coordinately expressed in mandibular prominences prior to and during tooth formation using reverse transcriptase-polymerase chain reaction (RT-PCR). During the dental lamina stage, ligand and receptor were present in both enamel organ epithelium and adjacent mesenchymal cells. During the bud stage, ligand and receptor were localized mainly to the enamel organ epithelium. Exogenous PDGF-A at 20 ng/ml enhanced tooth development to reach the cap stage with increased tooth size (P < 0.05) using embryonic day (E)10 mandibular explants cultured in serumless, chemically defined medium. A significant increase in DNA synthesis was observed within enamel organ epithelium at E10+4 when the mandibular explants were treated with PDGF-A at 20 ng/ml. These data suggest that PDGF-A and its cognate receptor (PDGFR-alpha) regulate the size and stage of tooth development via an autocrine mechanism during odontogenesis in vitro. PMID- 9853971 TI - Clinical pharmacology and therapeutic drug monitoring of zonisamide. AB - Zonisamide (1,2-benzisoxazole-3-methanesulfonamide) is a new antiepileptic drug developed in Japan. This compound is insoluble in water, and it is available in tablet and powder form. In experimental animals, this compound has been found to have a strong inhibitory effect on convulsions of cortical origin because it suppresses focal spiking and the spread of secondary generalized seizures. In humans, a series of double-blind, placebo-controlled studies revealed the efficacy of zonisamide for patients with refractory partial seizures and for selected patients with infantile spasms. Its antiepileptic mechanism of action remains unclear, but it is likely to involve blockade of both sodium and T-type calcium channels. Oral bioavailability of zonisamide is excellent in healthy human volunteers. Zonisamide is slowly absorbed and has a mean tmax of 5 to 6 hours. Almost 100% of it is absorbed; there is no difference in bioavailability between tablets and powder. Zonisamide concentrations are highest in erythrocytes and then in whole blood and plasma. It is approximately 40% to 60% bound to plasma proteins, primarily albumin. Its volume distribution is 0.9 to 1.4 L/kg. In adults, the elimination half-life is between 50 and 62 hours, and it takes as long as 2 weeks to reach steady state. The dose-serum level correlation is linear up to doses of 10 to 15 mg/kg per day, and the therapeutic range is 10 to 40 microg/ml. However, the relationship between serum zonisamide levels, clinical response, and adverse effects appears weak. Concurrent enzyme-inducing anticonvulsants such as phenytoin, carbamazepine, or barbiturates stimulate zonisamide metabolism and decrease serum zonisamide levels at steady state. Although zonisamide has been reported to increase the serum levels of phenytoin and carbamazepine in some patients, the interactions of zonisamide with other antiepileptic drugs seem to be of minor clinical relevance. A pilot study of zonisamide suppositories revealed that it is beneficial for patients with neurologic disorders in whom antiepileptic drugs cannot be administered by mouth. PMID- 9853972 TI - Delivery of therapeutic drug monitoring services: survey of Australasian clinical pharmacology laboratories. AB - Therapeutic drug monitoring (TDM) services, like many diagnostic services in public hospitals, have been under scrutiny over recent years as funding has decreased. The Australasian Society of Clinical and Experimental Pharmacologists and Toxicologists (ASCEPT) sought to review clinical pharmacology departments to consider how the changes that have been implemented have affected the delivery of TDM in recent years. A questionnaire was sent to such departments and all those known to be delivering TDM services responded. The survey demonstrated that of the 11 departments contributing TDM assays, 10 had lost tests and staffing to general biochemistry departments; eight departments had been delivering research and development in TDM. The TDM tests retained in clinical pharmacology were typically the more complex chromatographic or labor-intensive toxicology tests or the more expensive immunoassays. If this direction in Australasia is typical of the situation internationally, it should be a matter of great concern to all those with a particular interest in TDM. Is the future of TDM to be one in which only rapid immunoassays will be provided, and by a staff not fully able to provide pharmacokinetic support and interpretation of such tests (i.e., to become simply number-generating services) despite all the pharmacoeconomic data that is increasingly available? PMID- 9853973 TI - Therapeutic drug monitoring of tacrolimus in liver transplantation, phase III FK506 multicenter Spanish Study Group: a two-year follow-up. AB - The aim of the Multicentric Liver Transplant Spanish Study was to evaluate tacrolimus therapy at the reduced, initial oral dose of 0.1 mg/kg per day to maintain the immunosuppressive potency of the drug and to avoid toxicity. The dosage of tacrolimus (D), the trough blood concentrations (C), and the evolution of the ratio (D/C) were followed up for 2 years after transplantation in 50 adult patients (38 men, 12 women) undergoing liver allograft transplantation. A total of 1732 samples were analyzed using the IMx tacrolimus method. The overall mean+/ SD concentrations were 10.84 ng/ml+/-5.32 ng/ml. During the first month, the median of the tacrolimus levels was 8.40 ng/ml, and 73.1% of the analyzed samples were within the established therapeutic range. The median oral tacrolimus dose was progressively reduced from 0.12 mg/kg per day during the first month to 0.058 mg/kg per day at the end of study period. A significant negative association was observed between the ratio of D/C and the post-transplantation period (r=-0.3624; p < 0.001). The median D/C ratio ranged from 0.0144 at the end of the first month to 0.0053 at 1 year. Significant declines in D/C were observed after the first and the third months after transplantation. The decrease in corticosteroid doses and the increase in serum albumin may explain the reduction in clearance with time. PMID- 9853974 TI - Population pharmacokinetics of nitroglycerin and of its two metabolites after a single 24-hour application of a nitroglycerin transdermal matrix delivery system. AB - The purpose of this study was to assess the ability of our previously constructed pharmacokinetic (PK) model to describe nitroglycerin (GTN), 1,2-dinitroglycerin (1,2-GDN), and 1,3-dinitroglycerin (1,3-GDN) plasma concentrations after a single dose application of a GTN transdermal matrix delivery system. GTN, 1,2-GDN, and 1,3-GDN plasma concentrations were simultaneously fitted using a first-pass, mixed-order release, one-compartment PK model. Population PK parameter values were derived using an iterative two-stage methodology (IT2S). Some of the mean PK parameters estimates and their interindividual variability (CV%) were the percentage of the delivered GTN dose reaching the systemic circulation released by a first-order process A, 53% (44); the 1,2-GDN and 1,3-GDN formation rate constants, k(f1)9 h(-1) (67) and k(f2) 0.5 h(-1) (38), respectively; the metabolite elimination rate constant, k(m) 1 h(-1) (27); GTN, 1,2-GDN, and 1,3 GDN volumes of distribution (Vc/F 6 L [45]), V2/F 78 L [51]), and V3/F 29 L [40]), respectively). Mean calculated elimination half-lives (t1/2+/-standard deviation [SD]) for GTN and the GDN metabolites were 7+/-4 minutes and 33+/-7 minutes, respectively. The proposed PK model fitted the observed plasma concentrations of GTN, 1,2-GDN, and 1,3-GDN very well. This new transdermal matrix delivery system appears to behave pharmacokinetically in the same manner as a transdermal reservoir delivery system (Transderm-Nitro, Ciba-Geigy, Mississauga, Canada). PMID- 9853975 TI - Population pharmacokinetics of vancomycin in Japanese pediatric patients. AB - The population pharmacokinetic profile of vancomycin (VCM) in Japanese pediatric patients infected with methicillin-resistant Staphylococcus aureus was analyzed using 181 samples of serum concentration data from 49 patients obtained in routine drug monitoring. The one-compartment linear model was adopted, where the VCM clearance (CL) and the distribution volume (Vd) were correlated with covariates such as postnatal age (AGE) and body weight (BWT). The population pharmacokinetic analysis program NONMEM with the first-order conditional estimation method was used. The results showed that the population mean clearance normalized by BWT increases with AGE up to 1 year of age [CL(L/hour per kg) = 0.1 19 + 0.0619 x (AGE - 1)] and decreases with age over 1 year old [CL(L/hour per kg) = 0.119 + 0.00508 x (1 - AGE)]. The population mean of the distribution volume normalized by BWT was independent of AGE (Vd (L/kg) = 0.522). The interindividual variability of CL was 39.6%, and that of Vd was 18.8%. The intraindividual, residual variability was 34.6%. These results were compared with those in other articles, and a guideline for dosage adjustment in VCM therapy is discussed. PMID- 9853976 TI - Comparison of total and free phenytoin serum concentrations measured by high performance liquid chromatography and standard TDx assay: implications for the prediction of free phenytoin serum concentrations. AB - The free fraction of phenytoin (PHT) in serum increases considerably in combination with valproic acid (VPA), depending on the VPA concentration. Equations to predict the free PHT concentration (PHTf) from the total PHT concentration (PHTt) and from the VPA concentration were developed by Haidukewych and colleagues. (equation 1: PHTf = 0.095 x PHTt + 0.001 x VPA x PHTt) and May and colleagues (equation 2: PHTf = 0.0792 x PHTt + 0.000636 x VPA x PHTt]; in both equations, PHTf, PHTt, and VPA are given in microg/ml. Obviously, the equations give different predictions. The aim of this study was to investigate whether different methods for the determination of PHTt and PHTf were responsible for the differences; equation 1 was calculated from standard TDx measurements and equation 2 from high-performance liquid chromatography (HPLC) values. A total of 52 samples from patients with VPA (n=26) or without VPA (n=26) were analyzed using HPLC and TDx. The concentrations measured by HPLC and TDx were highly correlated but TDx yields significantly higher PHTt (Y = 0.98 x X + 2.46; X = HPLC, Y = TDx, r2 = 0.957) and, in particular, higher PHTf concentrations (Y = 1.03 x X + 0.30; X = HPLC, Y = TDx, r2 = 0.919), compared with our HPLC method. The accuracy of the predictive equations depends on the method used for the determination of PHTt and PHTf. The best predictions of PHTf were obtained if equation 2 and HPLC measurements were used. However, the differences in the predicted PHTf could only partly be explained by the different methods of determination. PMID- 9853977 TI - Relative biologic availability and pharmacokinetics of albuterol preparations in healthy Chinese. AB - The authors' goal was to study the biologic availability and pharmacokinetics of two different formulations of controlled-release (CR) tablets of albuterol. A two way, cross-over biologic availability study was performed with 20 healthy male volunteers to evaluate the relative biologic availability of two CR tablets of albuterol versus two different formulations of immediate-release (IR) albuterol tablets. Albuterol concentrations in plasma were measured using an HPLC procedure. Each patient subject received a 4.8-mg CR tablet every 12 hours for 6 days and a 4.8-mg IR tablet every 8 hours for 6 days. Tests of single doses and steady state were assayed for CR and IR albuterol tablets. The mean Cmax and tmax for two CR tablets given after a single dose were 7.3+/-1.5, 7.9+/-1.4 microg x L(-1) and 4.6+/-0.8, 4.8+/-0.5 hours, respectively. The Cmax and tmax for two IR tablets given after a single dose were 12.5-/+1.3, 15.3 +/-2.3 microg x L(-1) and 1.3+/-0.4, 1.6+/-0.4 hours, respectively. The relative biologic availability of two CR albuterol tablets were 106.0+/-6.2% and 109.8+/-9.0%, respectively. Administration of CR albuterol twice a day provides an alternative to administration of IR albuterol three or four times a day. PMID- 9853978 TI - Inducing effect of phenobarbital on clozapine metabolism in patients with chronic schizophrenia. AB - The steady state plasma concentrations of clozapine and its two major metabolites, norclozapine and clozapine N-oxide, were compared in patients with schizophrenia treated with clozapine in combination with phenobarbital (n=7), and in control patients treated with clozapine alone (n=15). Patients were matched for sex, age, body weight, and antipsychotic dosage. Patients comedicated with phenobarbital had significantly lower plasma clozapine levels than those of the controls (232+/-104 versus 356+/-138 ng/ml; mean, SD, p < 0.05). Plasma norclozapine levels did not differ between the two groups (195+/-91 versus 172+/ 61 ng/ml, NS), whereas clozapine N-oxide levels were significantly higher in the phenobarbital group (115+/-49 versus 53+/-31 ng/ml, p < 0.01). Norclozapine/clozapine and clozapine N-oxide/ clozapine ratios were also significantly higher (p < 0.001) in patients comedicated with phenobarbital. These findings suggest that phenobarbital stimulates the metabolism of clozapine, probably by inducing its N-oxidation and demethylation pathways. PMID- 9853979 TI - Modeling INR data to predict maintenance fluindione dosage. AB - This study was designed to construct a pharmacokinetic/pharmacodynamic model describing the evolution of International Normalized Ratio (INR) under oral anticoagulation treatment by fluindione in patients and to develop a method for individualization of fluindione dosage. Three indirect response models describing the concentration-INR relationship were tested using a nonparametric estimation method. INR was modelled as a quantity being produced and eliminated. According to a log-likelihood ratio test, the evolution of INR was best modelled as an inhibition of its elimination by fluindione. The selected model was evaluated in 24 additional patients with INR measurements (after 2, 3, 4, 6, and 10 doses). Using a Bayesian method with data until day 4, INR was correctly predicted for days 6 and 10. The population characteristics of fluindione were estimated, pooling the two groups of patients. A Bayesian method for individualization of dosage regimen was developed, based on a risk function for INR at steady state. Prescription rules for fluindione were derived using this method retrospectively on the 73 patients in this study. PMID- 9853980 TI - High single dose of mitoxantrone and cytarabine in acute non-lymphocytic leukemia: a pharmacokinetic and clinical study. AB - In a phase I-II study, the authors evaluated the intracellular pharmacokinetics, toxicity, and efficiency of a high dose of mitoxantrone given as first induction in acute non-lymphocytic leukemia. Twenty-two patients with previously untreated de novo ANLL were included and received 30 or 40 mg/m2 mitoxantrone on day 1 by intravenous infusion over 1 hour and 500 mg/m2 ara-C twice a day for 5 days. If there was no complete remission (CR), a second induction with ara-C, etoposide, and amsacrine was given. The CR rate after two courses with this regimen was 77%. Median duration of severe neutropenia was 18 days in the 30-mg/m2 group and 25 days in the 40-mg/m2 group. Two patients had fatal lung complications probably unrelated to mitoxantrone. A third patient had a possible mitoxantrone-induced reversible lung complication. In the leukemic cells, we found a high accumulation of mitoxantrone which, in contrast to the plasma concentration, remained stable during the 48 hours studied. Compared with previous results with 12 mg/m2 mitoxantrone, the AUC for intracellular concentrations versus time for the first 20 hours studied was increased by 150% to 0.638 nmol/mg cell protein x hour with 30 mg/m2 mitoxantrone and by 260% to 1.103 nmol/mg cell protein x hour with 40 mg/m2 mitoxantrone. In conclusion, a high dose of mitoxantrone results in a high intracellular exposure of the leukemic cells, which may be an advantage in improving survival of these patients. PMID- 9853981 TI - Prevention of tricyclic antidepressant adsorption loss with diethylamine during solvent evaporation. AB - The authors evaluated the adsorption loss of tricyclic antidepressants in analytical procedures with solvent extraction and evaporation. In standard procedures with the use of triple solvent extraction between alkalinized and acidified samples before chromatographic analysis, the adsorption loss was more significant with the demethylated metabolites. As much as 50% adsorption loss can occur; this irreversible loss can be accounted for entirely during the solvent evaporation step. Because of differential adsorption loss among parent drugs, metabolites, and internal standards, the analytical methods usually had wide within-day and day-to-day variations. The authors found that the addition of as little as 0.05% diethylamine to the extract before evaporation completely eliminated the adsorption loss of amitriptyline-nortriptyline, imipramine desipramine, and doxepin-desmethyldoxepin. with subsequent improvement in procedure performance. This simple modification can be adopted readily by all laboratories that use solvent extraction and subsequent chromatographic analysis of tricyclic antidepressants. PMID- 9853982 TI - Carbamazepine-10,11-epoxide in therapeutic drug monitoring. AB - Carbamazepine (CBZ) is widely used in the treatment of epilepsy, frequently in combination with other anticonvulsants. Its metabolite, carbamazepine-10,11 epoxide, is pharmacologically active and is increased with concurrent use of valproate and other anticonvulsants. This pharmacokinetic interaction may be particularly important because CBZ, its epoxide, phenytoin, and lamotrigine all act on fast voltage-dependent sodium channels. Over a 2-month period, routine serum requests for CBZ (n=47) (excluding known cases of overdose) were also analyzed for CBZ epoxide, phenytoin, and lamotrigine using a simultaneous high performance liquid chromatographic (HPLC) method. Valproate was measured using fluorescence polarization immunoassay (FPIA). With concurrent phenytoin and lamotrigine administration, there was a relative increase in CBZ epoxide and a significant decrease in the ratio of CBZ to epoxide (from more than 5 to 3). If valproate was also present, the concentration of parent and metabolite increased significantly, causing potential toxicity. Two patients in this latter group had significant clinical toxicity, with parent CBZ concentrations in the reference range; a third patient suffered from poor control of seizures. This study illustrates the importance of awareness of the contribution of active metabolites in therapeutic drug monitoring and raises questions about the role of the routine monitoring of such metabolites. PMID- 9853983 TI - Elevated free fosphenytoin concentrations in uremic sera: uremic toxins hippuric acid and indoxyl sulfate do not account for the impaired protein binding of fosphenytoin. AB - Fosphenytoin is a new phosphate ester prodrug of phenytoin. Impaired protein binding of phenytoin in uremia has been extensively documented, which prompted us to investigate the protein binding of fosphenytoin in uremic sera. Also studied was the role of uremic toxins hippuric acid and indoxyl sulfate as potential inhibitor of the protein binding of fosphenytoin because these compounds impair protein binding of phenytoin in uremia. Five serum pools were prepared from normal volunteers and five pools from patients with uremia. None of them received phenytoin. The normal serum pools were diluted with saline to mimic the albumin concentration of uremic pool. Both the diluted normal pool and the uremic pool were supplemented with fosphenytoin; after incubation at room temperature for 30 minutes, total and free fosphenytoin concentrations as phenytoin equivalents were measured using fluorescence polarization immunoassay (Abbott Laboratories; Abbott Park, IL, U.S.A.). The authors observed significantly elevated free fosphenytoin concentration in uremic sera compared with that of normal sera in all cases. Because both normal and uremic sera had the same concentrations of albumin, the elevated free fosphenytoin concentration in uremic sera was not caused by hypoalbuminemia. Both indoxyl sulfate and hippuric acid cause significant displacement of phenytoin from protein binding. In contrast, none caused any displacement of fosphenytoin from protein binding. PMID- 9853984 TI - Interference from digitoxin-like immunoreactive factors reduced in a new monoclonal chemiluminescent digitoxin assay. AB - Endogenous digoxin-like immunoreactive factors (DLIF) can interfere with some digoxin immunoassays. We looked for similar interference, called digitoxin-like immunoreactive factors (DTLIF) in two digitoxin immunoassays: A new chemiluminescent assay (CLIA), processed on the automated random access immunoassay system ACS:180, and a fluorescent polarization assay (FPIA), processed on the semiautomated TDx batch analyzer. One hundred thirty-seven samples of sera were tested from nondigitalized pregnant women, patients with liver or kidney diseases, and cord blood. The CLIA digitoxin assay uses a murine monoclonal antibody and requires no sample pretreatment; the FPIA digitoxin assay uses a polyclonal rabbit antibody and requires sample precipitation. Both assays have a similar dynamic range and sensitivity and give comparable results with commercial controls and external quality control survey samples. Although the CLIA detected no digitoxin in any sample tested, the FPIA showed apparent digitoxin concentrations of more than 2.0 ng/ml for 100% and 44% among cord blood and liver disease specimens, respectively. The highest DTLIF concentration was found in serum from a patient with liver disease (18.1 ng/ml). When spiked with 32 ng/ml digitoxin, six of the samples containing DTLIF generated FPIA digitoxin values of 6% to 27.5% more than the expected digitoxin levels. Two specimens with no detectable DTLIF activity were run as controls, and when spiked with digitoxin, showed target digitoxin concentrations in the FPIA. The CLIA recovered near the target digitoxin values (32 ng/ml) in all spiked samples. It was concluded that the polyclonal FPIA digitoxin assay may give discordant digitoxin concentrations in some patient groups because of interference from digitoxin-like immunoreactive factors. The CLIA digitoxin assay is not affected by DTLIF interference. PMID- 9853986 TI - Comparison of four immunoassays for the detection of lorazepam in urine. AB - Four healthy patient subjects were each given a single, 1-mg lorazepam tablet. Urine samples from all patient subjects were collected at 12 intervals (0-2, 2-5, 5-8, 8-11, 11-14, 14-24, 24-26, 26-29, 29-32, 32-35, 35-38, and 38-48 hours). An aliquot from each urine collection was screened using cloned enzyme donor immunoassay (CEDIA), enzyme-multiplied immunoassay technique (EMIT) II, EMIT dau, and fluorescence polarization immunoassay (FPIA) without and with hydrolysis using beta-glucuronidase. Using a 200 ng/mL calibrator cut-off, none of the four immunoassays gave a positive response before hydrolyzation of the urine samples. For offline hydrolysis using Helix pomatia beta-glucuronidase, 35, 3, 0, and 4 of 48 urine samples gave positive responses on the previously listed immunoassays. The CEDIA method also gave 32 of 48 positive responses for online hydrolysis using Escherichia coli beta-glucuronidase. Online hydrolysis can be conveniently automated by including the beta-glucuronidase in the first of the two reagents combined with the urine sample. PMID- 9853985 TI - Co-trimoxazole and stavudine interference in a high-performance liquid chromatographic analysis for didanosine in human plasma. AB - Recently, the authors were confronted with interference of stavudine and co trimoxazole when analyzing the antiretroviral drug didanosine (ddI) in plasma of HIV-1-infected patients using reverse-phase high-performance liquid chromatography with ultraviolet detection. After increasing the percentage of methanol in the mobile phase from 4% to 8% vol/vol and after decreasing the pH of the mobile phase from 6.8 to 5.8, the authors were able to separate didanosine from stavudine and co-trimoxazole (both are frequently used drugs in combination with didanosine). Subsequently, the adapted bioanalytic methodology was validated, and validation results showed that this new methodology can be used for the quantitative determination of didanosine in human plasma. This observation makes clear that combination therapy for human immunodeficiency virus with multiple (often chemically related) drugs has the potential of unexpectedly complicating bioanalytic analyses because therapeutic strategies may change rapidly after publication of a bioanalytic methodology. Thus, it is evident that the investigation of interference of potentially coadministered drugs should be a standard procedure during the development of any bioanalytical methodology in any laboratory. PMID- 9853987 TI - Multicenter comparison of first- and second-generation IMx tacrolimus microparticle enzyme immunoassays in liver and kidney transplantation. AB - Tacrolimus is a potent immunosuppressive drug successfully used for baseline and rescue immunosuppression in patients after liver and kidney transplantation. Data from several clinical trials have demonstrated the efficacy of tacrolimus in the prevention of allograft rejection, even at lower concentrations in the therapeutic range (5-15 microg/L). In fact, some patients with tacrolimus levels at less than 5 microg/L have excellent hepatic or kidney function. The limit of detection of the IMx Tacrolimus I assay (TAC I; Abbott Laboratories, IL) is only 5 microg/L and that of the lower tacrolimus calibrator is 10 microg/L. The second generation assay uses the same monoclonal antibody and the same IMx technology but offers improved sensitivity, with a dynamic range from 0 microg/L to 30 microg/L (lower calibrator, 3 microg/L). The aim of this multicenter study was to evaluate the new IMx Tacrolimus II assay (TAC II) by assessing its precision, sensitivity, performance, and correlation degree relative to the IMx TAC I assay. The study was performed at three centers in Spain. The within-run coefficients of variation (CVs) obtained for the new assay, using each of the trilevel controls in replicates of 20 during 3 consecutive days, were 8.06%, 4.38% and 5.09% at 5 microg/L, 11 microg/L, and 22 microg/L, respectively. The corresponding between run CVs obtained measuring each of the three controls in duplicate on 10 consecutive days were 9.54%, 6.38% and 5.75%. The limit of detection, with 97.5% confidence, was 1.22 microg/L. TAC II results (Y) were compared with those from the original TAC I assay (X) analyzing 293 whole blood samples from liver (n=145) and kidney (n=148) transplant recipients. The correlation study with patient samples (using the Passing-Bablock method) was y=1.056, x + 0.017, r=0.927. No statistically significant differences were observed between assays (TAC I versus TAC II) in the mean values obtained for total patients (9.89+/-5.42 microg/L versus 10.49+/-5.63 microg/L), liver patients (9.16+/-4.79 microg/L versus 10.00+/-5.20 microg/L), and kidney patients (10.62+/-5.87 micro g/L versus 10.98+/-5.99 microg/L). The new IMx TAC II assay demonstrated the same precision and accuracy that characterized the original assay but showed improved sensitivity to the demands of tacrolimus monitoring in the lower therapeutic range of drug concentrations. PMID- 9853988 TI - Simultaneous high-performance liquid chromatography analysis of azithromycin and two of its metabolites in human tears and plasma. AB - This article describes a high-performance liquid chromatographic (HPLC) method for the measurement of azithromycin (AZI) and two of its metabolites, 9a-N desmethylazithromycin (ADES) and N-desmethylazithromycin (NDES), in human tears and plasma. The drug, metabolites, and internal standard (n-propylazithromycin [IS]) were detected electrochemically after injection of the extracted sample into the HPLC system. The peak height ratio (AZI, ADES, or NDES to IS) varied linearly, with concentrations in the ranges of 0.1 mg/L to 2.0 mg/L (tears) and 0.01 mg/L to 2.0 mg/L (plasma) of AZI, ADES, and NDES; the correlation coefficient (r) was more than 0.994 mg/L for all of the compounds (n=6). The analysis of tear samples collected at different intervals within 12 hours to 144 hours after a dose of 20 mg/kg of AZI from a trachoma patient yielded concentrations ranging from 1.52 mg/L to 0.34 mg/L for AZI, 0.79 mg/L to 0.27 mg/L for ADES, and 1.99 mg/L to less than 0.20 mg/L for NDES. The concentration of AZI in plasma ranged from 0.15 mg/L to 0.01 mg/L, whereas ADES and NDES were undetectable. PMID- 9853989 TI - Evaluation of an immunoassay (EMIT) for mycophenolic acid in plasma from renal transplant recipients compared with a high-performance liquid chromatography assay. AB - Mycophenolic acid is an immunosuppressant administered as a bioavailable ester, mycophenolate mofetil. The pharmacokinetics of mycophenolic acid have been reported to be variable. Accurate measurement of concentrations of this drug could be important to adjust doses. The aim of this study was to compare the enzyme-multiplied immunoassay technique (EMIT [Dade Behring; San Jose, CA, U.S.A.]) for mycophenolic acid with a high-performance liquid chromatographic (HPLC) assay using samples collected from renal transplant recipients. The HPLC assay used solid phase extraction and a C18 stationary phase with ultraviolet (UV) detection (254 nm). The immunoassay required no manual sample preparation. Plasma samples (n=102) from seven patients, collected at various times after a dose, were analyzed using both methods. Both assays fulfilled quality-control criteria. Higher concentrations were consistently measured in patient samples when using EMIT. The mean (+/-standard deviation [SD]) bias (EMIT-HPLC) was 1.88+/-0.86 mg/L. The differences in concentrations were higher in the middle of a dosage interval, suggesting that a metabolite might have been responsible for overestimation. Measurement of glucuronide concentrations by HPLC demonstrated only a weak correlation between assay differences and glucuronide concentrations. If the crossreacting substance is active, EMIT could provide a superior measure of immunosuppression; if inactive, further work is needed to improve antibody specificity. In conclusion, it was found that EMIT overestimates the concentration of mycophenolic acid in plasma samples from renal transplant recipients compared with HPLC analysis. PMID- 9853990 TI - Plasma indomethacin assay using high-performance liquid chromatography electrospray-tandem mass spectrometry: application to therapeutic drug monitoring and pharmacokinetic studies. AB - The authors report the use of high-performance liquid chromatography-electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) for the quantification of indomethacin (IND) in plasma with microscale sample preparation. Plasma samples (100 microL) and mefanamic acid (internal standard [IS]), buffered to pH 3.5, were prepared using solid phase extraction and chromatographed using a C8 column. The mobile phase composition was 80% methanol to 20% ammonium acetate buffer (40 mM, pH 5.1). A flow rate of 300 microL per minute was used with a 1-to-12 postcolumn split into the mass spectrometer. Selected reaction monitoring with mass transitions m/z 357.9-->139.0 and m/z 242-->209.0 were used for IND and IS, respectively. The chromatographic analysis time was 4 minutes. The assay was linear from 5 microg/L to 2000 microg/L with interday imprecision (n=5) over the analytic range (5%). At four concentrations (10 microg/L, 25 microg/L, 250 microg/L, 1500 microg/L), assay imprecision was 9% (total coefficient of variation [CV]) and accuracy ranged between 96.5% and 102.8% (n=16). The absolute recovery of IND and IS was 74% (n=8) and 95% (n=24), respectively. This method was developed and validated in less than 10 working days, had a lower limit of quantification than reported HPLC-ultraviolet (UV) methods, and uses small sample volumes. These factors illustrate the power of HPLC-ESI-MS/MS for drug analysis. Furthermore, the ability of this method to measure IND over a wide concentration range makes it suitable for therapeutic drug monitoring and pharmacokinetic studies. PMID- 9853991 TI - High-performance liquid chromatography-electrospray tandem mass spectrometry for the measurement of 1-(3,4-dichlorobenzyl)-5-octylbiguanide in human serum. AB - 1-(3,4-dichlorobenzyl)-5-octylbiguanide (OPB-2045) is a new biguanide antimicrobial agent currently in clinical use as a topical bactericidal antiseptic. A method combining high-performance liquid chromatography (HPLC) with electrospray ionization (triple and quadruple stage) tandem mass spectrometry (ESI-MS/MS) was developed to quantify OPB-2045 in human serum. Solid phase extraction was performed on 0.2 ml of sample to ensure a high level of sensitivity before HPLC-ESI-MS/MS analysis. The limit of quantitation for the method was set at 0.05 ng/ml. Intra-assay and interassay precision were less than 13.7%, with a deviation from the expected value of no greater than 10.5% at a concentration range of 0.05 ng/ml to 5 ng/ml. Decomposition of OPB-2045 in human serum did not occur after storage for 15 months at -20 degrees C, even after three repetitions of freezing and thawing. Application of this method was demonstrated in a pharmacokinetic study of OPB-2045 in healthy patient subjects after a single topical application of 5 g/L preparation of its liquid formulation. PMID- 9853992 TI - Screening for the detection of angiotensin-converting enzyme inhibitors, their metabolites, and AT II receptor antagonists. AB - A gas chromatography-mass spectrometry (GC-MS) screening procedure was developed for the detection of angiotensin-converting enzyme (ACE) inhibitors, their metabolites, and angiotensin (AT) II receptor antagonists in urine as part of a systematic toxicologic analysis procedure for acidic drugs and poisons after extractive methylation. The part of the phase-transfer catalyst remaining in the organic phase was removed by solid phase extraction on a diol phase. The compounds were separated by capillary GC and identified by computerized MS in the full scan mode. Using mass chromatography with the ions m/z 157, 160, 172, 192, 204, 220, 234, 248, 249, and 262, the possible presence of ACE inhibitors, their metabolites, and AT II antagonists could be indicated. The identity of positive signals in such mass chromatograms was confirmed by comparison of the peaks underlying full mass spectra with the reference spectra recorded during this study. This method allowed detection of therapeutic concentrations of ACE inhibitors (benazepril, enalapril, perindopril, quinapril, ramipril, trandolapril, their metabolites, or both) and therapeutic concentrations of the AT II antagonist, valsartan, in human urine samples. Human urine samples were not available for testing cilazapril, moexipril, and losartan; they were detected only in rat urine. The overall recoveries of ACE inhibitors ranged between 80% and 88%, with a coefficient of variation (CV) of less than 10% and the limit of detection of at least 10 ng/ml (signal to noise ratio 3) in the full-scan mode. The overall recovery of the valsartan was 68%, with a CV of less than 10%; the limit of detection was at least 10 ng/ml (S/N 3) in the full scan mode. PMID- 9853993 TI - Sodium stibogluconate (pentostan) overdose in a patient with acquired immunodeficiency syndrome. AB - A 32-year-old man with acquired immunodeficiency syndrome (AIDS) admitted to the hospital for treatment of visceral leishmaniasis was inadvertently given 10 times the prescribed first dose of sodium stibogluconate ([Sb] 6.5 g instead of 0.65 g). He experienced no immediate major toxicity during the first 48 hours, but a significant rise of pancreatic enzyme activities was observed (amylase at 10 times the upper limit of normal, lipase at 50 times the upper limit of normal) without clinical signs or indications on computed tomography (CT) of pancreatitis. The third day after the overdose, he developed appendicitis, which appeared coincidental; he recovered uneventfully from surgery. Most of the overdose of Sb was eliminated within the first few hours. Pharmacokinetics remained linear; the rapid, long elimination half-lives (2.7 hours and 54 hours, respectively) were similar to those in previously published results. The administration of a chelating agent, dimercaptosuccinic acid (DMSA), 72 hours after the Sb overdose did not modify the pharmacokinetics of the medication. PMID- 9853994 TI - Is 10 milligrams selegiline essential as an adjunct therapy for the symptomatic treatment of Parkinson's disease? AB - Selegiline is used as an adjunct to levodopa in the symptomatic treatment of Parkinson's disease (PD). The normal daily dose of selegiline is 10 mg administered orally. This study, based on monoamine oxidase-B (MAO-B) inhibition, investigates whether a reduction in selegiline dose can provide the same beneficial effects seen with a 10-mg dose. The inhibition of platelet MAO-B activity against multiple dosing of selegiline (2.5, 5, and 7.5 mg) was predicted from the data obtained from literature (0.5, 1.0, 1.5, and 10 mg). A pharmacokinetic-pharmacodynamic model for selegiline was also developed. The data suggested that by 96 hours (four doses) the inhibition of platelet MAO-B activity is approximately 95% after a daily dose of 2.5 mg selegiline, whereas it takes only 48 hours (two doses) for doses of 5 mg and 7.5 mg to achieve this degree of inhibition. The pharmacokinetic-pharmacodynamic model was best described by a sigmoidal Emax model with an effect compartment. Based on the inhibition of MAO-B activity, a reduction in daily oral dose of selegiline appears possible without compromising the therapeutic effect. Therefore, lower doses of selegiline should be tested in clinical trials. PMID- 9853995 TI - Current bioequivalence criteria are adequate for oral cyclosporin A formulations. PMID- 9853996 TI - Oral intake of perphenazine decanoate. PMID- 9853997 TI - Cryo/Cuff use after arthroscopic surgery: effect on knee joint temperature. AB - Any modality that alters joint homeostasis could have a potential negative effect on cartilage. During arthroscopic knee surgery, the temperature in the knee joint decreases significantly. The use of cryotherapy after this surgical procedure could maintain or increase these temperature variations. This prospective, randomized study evaluated the change in intra-articular knee temperature with the use of the Cryo/Cuff (Aircast Inc, Summit, New Jersey) in 30 patients after arthroscopic knee surgery. In 15 patients, a Cryo/Cuff with an automatic pump was applied immediately after surgery (group 1), and in the remaining 15 patients, only a wound dressing was applied (group 2). The mean intra-articular temperature at the beginning of the procedure was 34.8+/-1 degrees C for both groups. At the end of surgery, the temperature was 26.8+/-2.2 degrees C for group 1 and 27.5+/ 2.2 degrees C for group 2. One hour after the end of the procedure, the mean temperature was 34+/-1.6 degrees for group 1 and 34.8+/-1.7 degrees C for group 2. These differences were not statistically significant. These results demonstrated that the use of the Cryo/Cuff does not alter temperature recovery in the knee, thereby excluding potential damage to the articular cartilage. PMID- 9853998 TI - In vitro and in vivo fibrochondrocyte growth behavior in fibrin gel: an immunohistochemical study in the rabbit. AB - This study investigated matrix formation of rabbit meniscal fibrochondrocytes synthesized in vitro and the viability of fibrochondrocytes encapsulated in fibrin gel after transplantation into rabbit meniscal lesion sites in vivo. The matrix of newly formed meniscal fibrochondrocytes encapsulated in fibrin gel was evaluated using immunohistochemical methods. The meniscal cells in the three dimensional encapsulating fibrin gel proliferated well for a 2-month period. Examination of the in vivo transplantation confirmed that the newly formed cartilage produced S-100 protein. These observations suggest that meniscal fibrochondrocytes encapsulated in fibrin gel are suitable for cell division and matrix production, and the use of fibrin gel as a scaffolding delivery substance may be beneficial for the transplantation of viable meniscal fibrochondrocytes in resurfacing meniscoplasty. PMID- 9853999 TI - Caudal slope of the tibia and its relationship to noncontact injuries to the ACL. AB - To determine the role that an increased caudal slope of the tibia might have on the incidence of anterior cruciate ligament (ACL) injury, tibial slope was measured in 49 patients (50 knees), with noncontact ACL injury mechanisms (group 1) and an age-matched group of 39 patients (50 knees) with a diagnosis of patellofemoral pain syndrome (group 2). No significant difference was noted in mean posterior slope between the two groups (group 1: 9.7+/-1.8 degrees and group II: 9.9+/-2.1 degrees) or after controlling for bilateral patients in two separate trials. These results indicate that increased caudal tibial slope does not appear to be a risk factor in the creation of noncontact injuries to the ACL. PMID- 9854000 TI - Osteochondritis dissecans of the knee. AB - Osteochondritis dissecans is a separation of an articular cartilage subchondral bone segment from the remaining articular surface. This is a different entity from osteonecrosis, with which osteochondritis dissecans is commonly confused. In osteochondritis dissecans, the fragment separates from a vascular normal bony bed, while in osteonecrosis, the fragment typically is more peripheral and separates from an avascular bony bed. Osteochondritis dissecans is more common in adolescents and young adults, with the knee, elbow, and ankle being the most common sites. Injury to an area of fairly tenuous blood supply is the most likely cause. Treatment is typically nonoperative for stable lesions and operative for unstable lesions. Most patients do well with no long-term sequelae, but this depends on a variety of factors including the location and size of the lesion, patient age, and treatment. PMID- 9854001 TI - Septic knee arthritis secondary to a functional brace after ACL reconstruction. PMID- 9854002 TI - Long-term follow-up of bilateral steroid-induced osteonecrosis of the lateral femoral condyles in a patient with Crohn's disease. PMID- 9854003 TI - Evaluation and management of tumors about the knee. PMID- 9854004 TI - Benign bone and soft-tissue tumors about the knee. PMID- 9854005 TI - Malignant tumors about the knee. AB - The knee is the most common location for malignant bony tumors as well as a number of soft-tissue sarcomas. The clinical presentation of these lesions usually involves pain and swelling, often associated with a palpable mass. This can occur rapidly or take a more indolent course. Proper evaluation and staging prior to biopsy of these tumors is necessary. This should be done by a surgeon experienced in the treatment of these tumors and at a facility where a multispecialty team can provide patient care. The primary surgical choices for treating malignant musculoskeletal tumors are limb salvage and amputation. One of the major difficulties in performing limb salvage about the knee is the lack of soft-tissue coverage after resection. Local or even free flaps are often necessary. Additionally, reattachment of the extensor mechanism is necessary. These problems and the use of allografts often lead to a high incidence of complications including infection, wound breakdown, allograft fracture, or extensor mechanism problems with resultant poor functional result. PMID- 9854006 TI - [Service " conseillers GBEA "] PMID- 9854007 TI - [Precarious situation of obstetric practice in Gabon]. AB - In 1993, 2,008 deliveries were recorded at the Provincial Maternity Hospital at Franceville in Gabon. The frequency of cesarean section was 3%. The perinatal mortality of children born by cesaraean section was high, 213 per thousand. The principal indications for cesarean section were the baby being too large to pass through the pelvis, bicicatricial uterus, breech presentation and toxemia during pregnancy. The maternal mortality rate was 200 per 100,000, similar to the rates of most African countries, and 75% of the women that died had undergone cesarean section. The mortality rate for cesarean section was high (4.9%), so the indications for cesarean section in underdeveloped countries are limited. Malaria was the principal reason for the hospitalization of pregnant women, because it is endemic and is a serious condition for pregnant women. The next most frequent causes of hospitalization were a high risk of premature labor and hyperemesis gravidarum, the frequency of which is high among pregnant African women, particularly those of West Africa. Toxemia in pregnancy was the fourth most important cause of hospitalization. The rate of cesarean section rupture was 2.5 per thousand. Only 20% of these cases involved a cicatricial uterus, with no maternal deaths but a fetal mortality rate of 100%. The frequency of premature birth was 4.23% and the perinatal mortality rate was 48 per thousand, with 37 stillbirths per thousand and an early neonatal mortality rate of 11 per thousand. The perinatal mortality of breech presentations was high (330 per thousand), with 13.9 delivered by cesarean section. These levels are similar to those for other African countries. Maternal health could be improved by introducing several consultations during pregnancy, improving hospital hygiene and making antibiotics more widely available. Fetal survival could be improved by preventing premature births, providing more help with delivery, decreasing the time to intervention and improving neonatal resuscitation techniques. PMID- 9854008 TI - [Bacteriological quality of spring and well water in Yaounde (Cameroon)]. AB - The drinking water supply in Yaounde, Cameroon, often fails to meet demand and this leads many people to seek water from springs and wells the microbiological quality of which is unknown. We carried out a microbiological survey of five spring water points and ten wells. The fifteen study sites were located in various residential districts and were selected according to geographical location and number of users. Microorganisms were isolated on specific culture media using the membrane filter technique. We found that these water supplies contained many types of bacteria including Pseudomonas aeruginosa, Aeromonas hydrophila and indicators of fecal contamination. Maximum annual concentrations differed at the various sites and over time, from 750 to 98,000 CFU/l for P. aeruginosa, 40 to 78,000 CFU/l for A. hydrophila and 300 to 58,000 CFU/l for fecal indicator bacteria. The density of fecal bacteria was strongly correlated (p < 0.01) with those of P. aeruginosa in spring water and A. hydrophila in well water. The degree of correlation between microbial concentrations differed between sites. The source of fecal contamination differed between sites and was animal, human or both. The densities of bacteria recorded were high. Therefore, consumers of water drawn from these sources are at short-term risk of contracting waterborne diseases. PMID- 9854009 TI - [Idiopathic thrombopenic purpura in a child in black Africa: a case report in Togo]. AB - Idiopathic thrombocytopenic purpurea (ITP) is an autoimmune disease that occurs frequently in Europe and the US, but has rarely been described in Africa. Case report. An 8-year-old girl was admitted for cutaneous and mucosal bleeding. She had a low platelet count (11 x 10(9)/l). ITP was suspected and the diagnosis was confirmed by bone marrow examination. Corticosteroid treatment was effective. DISCUSSION: This is the second case reported in Togo since 1982. The diagnosis of the disease is straightforward, so the lack of cases reported in central Africa suggests that the disease is rare in this region. The rarity of the disease may be due to genetic or environmental factors, or it may simply be that physicians overlook this disease when making their diagnosis. Corticosteroids are now the preferred treatment for ITP because of the risk of transmitting Creutzfeldt Jacob's disease by intravenous administration of immunoglobulin. Splenectomy is the ultimate treatment for chronic forms. CONCLUSION: Unlike other diseases, the diagnosis and treatment of which require methods unavailable in parts of Africa, ITP treatment, as currently practiced in countries of the northern hemisphere, is within the reach of most African countries. Further studies are required to determine the true frequency of the disease in central Africa. PMID- 9854010 TI - [Diabetes mellitus in sub-saharan Africa]. AB - Diabetes mellitus is becoming more common in African cities, where it may affect up to 7% of the hospital population. It particularly affects poor male patients and 73 to 80% of those affected have non insulin-dependent diabetes. The frequency of non-obese, poorly cetogenic patients is high in Sub-Saharan Africa. This may be due to malnutrition, with a deficit either in protein or in calories. Such malnutrition is a major public health problem affecting children in Sudanese and Sahelian areas and may interact with environmental and genetic factors. In equatorial environments, the toxic effects of alcohol abuse on the pancreas are simply another environmental factor, reducing the endocrine function of the pancreas. These observations are important because: 1) diabetes mellitus has a severe social impact in this area and 2) nutrition has a general effect on the pathogenesis of diabetes mellitus. PMID- 9854011 TI - [Prediction and prevention of malaria epidemics in the valley of the Senegal River]. AB - The Sahel region has been suffering from severe drought for the last thirty years, with large deficits and a high degree of variability in the amount of annual rainfall. Agricultural production in the Sahelian zone of the Senegal River valley depends on the flooding of the river. The management of this flooding affects malaria transmission. The area is prone to malaria epidemics because the immunity level of the population is low. We studied epidemiological, meteorological and river level data to identify epidemic risk factors. We propose an epidemiological and managerial system for the early detection of risks and early intervention. This system is based mainly on the water level of the Senegal River and the early detection of unusual increases in the number of cases. PMID- 9854012 TI - [Types of hypertension in pregnant women of Benin admitted to the National University Hospital of Cotonou]. AB - We carried out a retrospective and prospective study between January 1995 and August 1996, of pregnant women with high blood pressure. The aim of this work was to determine the prevalence of each type of hypertension according to the classification of the American College of Obstetricians and Gynecologists (ACOG) and to evaluate the prognosis for the mother and child. The prevalence of hypertension in pregnancy was found to be 7.65%. A family history of hypertension, obesity and a personal history of hypertension in pregnancy were all risk factors. Severe hypertension (diastolic blood pressure (DBP) > 110 mm Hg) affected 59.4% of the women. Chronic hypertension occurred in 41.51% of cases, preeclampsia in 26.41% of cases, associated preeclampsia in 18.87% of cases and isolated hypertension in 13.21% of cases. Eclampsia (70.6%) was the principal maternal complication in this study population. Fifty-four of the women gave birth to normal babies, 21 to hypotrophic babies, 15 gave birth prematurely and 3 had miscarriages. Six of the seven deaths involved women with DBP above 110 mm Hg. PMID- 9854013 TI - [Families of antihypertensive drugs and blood pressure exercise profiles in black Africans at the National University Hospital of Cotonou]. AB - The study population consisted of ninety patients between the ages of 22 and 64 years. All had mild or moderate hypertension at rest and were subjected to exercise tests using a cycloergometer. The sex ratio for the patients was 0.875. Blood pressure and heart rate were recorded at three time points, at three minute intervals. The initial load was thirty watts for men and 25 watts for the women. A graph of changes in blood pressure with exercise was produced for each type of antihypertensive drug or combination of drugs. At least nine patients were treated with each type of drug or combination, and blood pressure and heart rate were recorded during exercise to produce the exercise blood pressure profile. Healthy adults generate curves that lie between Y'1 = 0.5X + 108.43 and Y"1 = 0.34X + 83.95 [2]. Any patient generating a curve lying between these two limits was regarded as having a normal exercise blood pressure profile. This study identified two groups of hypertension treatments with respect to exercise blood pressure profile. Abnormal exercise blood pressure profiles were obtained with treatments involving inhibition of calcium receptor activity, diuretics and central antihypertensive drugs. Normal exercise blood pressure profiles were obtained with treatments involving the inhibition of adrenoreceptor activity, alone or in conjunction with a diuretic, and with combined central antihypertensive drug and diuretic treatment. PMID- 9854014 TI - [Clinical development of a new inactivated hepatitis A vaccine]. AB - An attenuated vaccine against hepatitis A was developed from the GBM strain of the virus, cultured on human diploid MRC5 cells. Each dose contained 160 antigen units, inactivated by formalin and adsorbed onto 0.3 mg aluminum hydroxide, in a volume of 0.5 ml. Intramuscular injection of the vaccine conferred immunity to hepatitis A, with antibody titers greater than those obtained by passive immunization with immunoglobulin. In clinical studies, immunocompromised subjects became immune shortly after the first injection and more than 90% were found to be protected after 14 days (titer above 20 mIU/ml by RIA). All subjects (100%) were protected one month after the first injection. Immunity persisted for at least six months and was strengthened by a booster injection. The antibody titers determined after the first booster injection were consistent with a projected period of protection of ten years. Adverse reactions were mild and occurred within the first few days after vaccination, with the patient usually recovering spontaneously. The most common reaction was mild local pain, usually associated with redness of the skin. A nodule was observed at the injection site in a small number of cases. Mild fever, asthenia, headache, myalgia/arthralgia and gastrointestinal tract disorders were also reported. Reactions were reported less frequently after the booster injection than after the initial dose. The vaccine was as well tolerated by patients seropositive for the hepatitis A virus as by seronegative subjects. Thus, this vaccine can be used for active immunization against hepatitis A in adults and adolescents. It can be administered as primary immunization and as a booster injection. PMID- 9854015 TI - [Cesarean sections in Senegal: coverage of needs and quality of services]. AB - A prospective longitudinal study was carried out of all women undergoing Cesarean section in the surgical maternity hospitals of Senegal between January 1 and December 31 1996. The epidemiology and quality of Cesarean sections were investigated. For each case, the following data were recorded: marital status, prenatal monitoring, conditions of hospitalization, indications for and outcome of surgery, maternal and neonatal follow-up one month after the operation. 2,436 Cesarean sections were performed. Of these, 2,269 cases were indexed and filed and 1,612 received a postnatal checkup one month after the operation. The mean age of the women involved was 26 years. The referral system is not effective, with 58% of patients being rushed to a surgical maternity unit in medically unsuitable forms of transport. Cesarean section is not widely available either geographically or economically. The mean national rate of Cesarean section was 0.6% of expected births but there were differences between regions. The main indications for Cesarean section were the fetus being too large to pass through the pelvic girdle (30%) and fetal suffering (18%). The maternal mortality rate was 3% and one third of the women who died did so immediately after the operation. Maternal morbidity occurred in 10% of cases, mainly due to postoperative infection. The rate of perinatal stress was 25%, most deaths being caused by neonatal distress (33%) or infection (18%). Thus, overall, both the availability and quality of Cesarean section in Senegal are poor. PMID- 9854016 TI - [International foreign bibliographic databases Medline and Internet Grateful Med V2.6. Presentation and how to use]. AB - This paper aims to provide a practical kit in French for reference retrieval from the Internet using Medline and its specific software interface, Internet Grateful Med V2.6. This reference retrieval tool is now available to all end users via the Internet. We explain here how to use it, screen by screen and button by button. PMID- 9854018 TI - Groucho/transducin-like enhancer of split (TLE) family members interact with the yeast transcriptional co-repressor SSN6 and mammalian SSN6-related proteins: implications for evolutionary conservation of transcription repression mechanisms. AB - The yeast proteins TUP1 and SSN6 form a transcription repressor complex that is recruited to different promoters via pathway-specific DNA-binding proteins and regulates the expression of a variety of genes. TUP1 is functionally related to invertebrate and vertebrate transcriptional repressors of the Groucho/transducin like Enhancer of split (TLE) family. The aim was to examine whether similar mechanisms underlie the transcription repression functions of TUP1 and Groucho/TLEs by determining whether TLE family members can interact with yeast SSN6 and mammalian SSN6-like proteins. It is shown in the present work that SSN6 binds to TLE1 and mediates transcriptional repression when expressed in mammalian cells. Moreover, TLE1 and TLE2 interact with two mammalian proteins related to SSN6, designated as the products of the ubiquitously transcribed tetratricopeptide-repeat genes on the Y (or X) chromosomes (UTY/X). These findings suggest that mammalian TLE and UTY/X proteins may mediate repression mechanisms similar to those performed by TUP1-SSN6 in yeast. PMID- 9854019 TI - Calsequestrin: more than 'only' a luminal Ca2+ buffer inside the sarcoplasmic reticulum. AB - In striated muscle, the sarcoplasmic reticulum (SR) Ca2+ release/ryanodine receptor (RyR) channel provides the pathway through which stored Ca2+ is released into the myoplasm during excitation-contraction coupling. Various luminal Ca2+ binding proteins are responsible for maintaining the free [Ca2+] at 10(-3)-10(-4) M in the SR lumen; in skeletal-muscle SR, it is mainly calsequestrin. Here we show that, depending on its phosphorylation state, calsequestrin selectively controls the RyR channel activity at 1 mM free luminal [Ca2+]. Calsequestrin exclusively in the dephosphorylated state enhanced the open probability by approx. 5-fold with a Hill coefficient (h) of 3.3, and increased the mean open time by about 2-fold, i.e. solely dephosphorylated calsequestrin regulates Ca2+ release from the SR. Because calsequestrin has been found to occur mainly in the phosphorylated state in the skeletal-muscle SR for the regulation of RyR channel activity, the dephosphorylation of calsequestrin would appear to be a quintessential physiological event. PMID- 9854017 TI - Nutrient and hormonal regulation of pyruvate kinase gene expression. AB - Mammalian pyruvate kinase (PK), a key glycolytic enzyme, has two genes named PKL and PKM, which produce the L- and R-type isoenzymes by means of alternative promoters, and the M1-and M2-types by mutually exclusive alternative splicing respectively. The expression of these genes is tissue-specific and under developmental, dietary and hormonal control. The L-type isoenzyme (L-PK) gene contains multiple regulatory elements necessary for regulation in the 5' flanking region, up to position -170. Both L-II and L-III elements are required for stimulation of L-PK gene transcription by carbohydrates such as glucose and fructose, although the L-III element is itself responsive to carbohydrates. The L II element is also responsible for the gene regulation by polyunsaturated fatty acids. Nuclear factor-1 proteins and hepatocyte nuclear factor 4, which bind to the L-II element, may also be involved in carbohydrate and polyunsaturated fatty acid regulation of the L-PK gene respectively. However, the L-III-element-binding protein that is involved in carbohydrate regulation remains to be clarified, although involvement by an upstream stimulating factor has been proposed. Available evidence suggests that the carbohydrate signalling pathway to the L-PK gene includes a glucose metabolite, possibly glucose 6-phosphate or xylulose 5 phosphate, as well as phosphorylation and dephosphorylation mechanisms. In addition, at least five regulatory elements have been identified in the 5' flanking region of the PKM gene up to position -279. Sp1-family proteins bind to two proximal elements, but the binding of proteins to other elements have not yet been clarified. Glucose may stimulate the transcription of the PKM gene via hexosamine derivatives. Sp1 may be involved in this regulation via its dephosphorylation, although the carbohydrate response element has not been determined precisely in the PKM gene. Thus glucose stimulates transcription of the PKM gene by the mechanism which is probably different from the L-PK gene. PMID- 9854020 TI - Diminished expression of phosphatidylethanolamine N-methyltransferase 2 during hepatocarcinogenesis. AB - Phosphatidylethanolamine N-methyltransferase (PEMT) is a liver-specific enzyme that converts phosphatidylethanolamine into phosphatidylcholine. At least two forms of PEMT are present in hepatocytes. However, PEMT activity is negligible in two hepatoma cell lines. Previous studies have indicated an inverse relationship between the expression of one form, PEMT2, and the rate of liver growth, suggesting that this enzyme might be involved in inhibition of hepatocyte proliferation. We have now investigated the expression of PEMT2 at various stages of hepatocarcinogenesis induced by chemical carcinogens. Expression of PEMT2 protein was decreased in liver samples that contained the first detectable proliferative lesions. At later stages of carcinogenesis, PEMT2 expression was obliterated. PEMT activity decreased, the levels of PEMT2 mRNA decreased and there was an increase in the activity of CTP:phosphocholine cytidylyltransferase, a key regulatory enzyme in the CDP-choline pathway of phosphatidylcholine biosynthesis. Southern blot analyses of restriction fragments of DNA showed no changes in the PEMT gene in hepatocarcinoma compared with normal liver. A role for PEMT2 in the control of hepatocyte proliferation remains an intriguing possibility. PMID- 9854021 TI - A conserved motif in the yeast nucleolar protein Nop2p contains an essential cysteine residue. AB - Nop2p is an essential nucleolar protein in Saccharomyces cerevisiae that is involved in large ribosomal subunit assembly. It has substantial homology with human p120, the proliferation-associated nucleolar antigen that is overexpressed in many human cancers. A motif containing an invariant Pro-Cys dipeptide is found in Nop2p, p120 and the bacterial Fmu proteins. A total of nine conserved residues, including Pro423 and Cys424, were individually altered in Nop2p by site directed mutagenesis. Nop2p function was abolished by conversion of Cys424 into either alanine or serine. All of the other Nop2p mutations tested sustained yeast viability, including glycine replacement of Pro423 and the conversion of a second conserved cysteine into alanine. The crucial role of Cys424 in Nop2p is intriguing, due to the critical roles that cysteine residues adjacent to a proline have in a number of nucleotide-modifying enzymes. PMID- 9854022 TI - Catalytic triad of microsomal epoxide hydrolase: replacement of Glu404 with Asp leads to a strongly increased turnover rate. AB - Microsomal epoxide hydrolase (mEH) belongs to the superfamily of alpha/beta hydrolase fold enzymes. A catalytic triad in the active centre of the enzyme hydrolyses the substrate molecules in a two-step reaction via the intermediate formation of an enzyme-substrate ester. Here we show that the mEH catalytic triad is composed of Asp226, Glu404 and His431. Replacing either of these residues with non-functional amino acids results in a complete loss of activity of the enzyme recombinantly expressed in Saccharomyces cerevisiae. For Glu404 and His431 mutants, their structural integrity was demonstrated by their retained ability to form the substrate ester intermediate, indicating that the lack of enzymic activity is due to an indispensable function of either residue in the hydrolytic step of the enzymic reaction. The role of Asp226 as the catalytic nucleophile driving the formation of the ester intermediate was substantiated by the isolation of a peptide fraction carrying the 14C-labelled substrate after cleavage of the ester intermediate with cyanogen bromide. Sequence analysis revealed that one of the two peptides within this sample harboured Asp226. Surprisingly, the replacement of Glu404 with Asp greatly increased the Vmax of the enzyme with styrene 7,8-oxide (23-fold) and 9, 10-epoxystearic acid (39 fold). The increase in Vmax was paralleled by an increase in Km with both substrates, in line with a selective enhancement of the second, rate-limiting step of the enzymic reaction. Owing to its enhanced catalytic properties, the Glu404-->Asp mutant might represent a versatile tool for the enantioselective bio organic synthesis of chiral fine chemicals. The question of why all native mEHs analysed so far have a Glu in place of the acidic charge relay residue is discussed. PMID- 9854023 TI - Phenol sulphotransferase SULT1A1 polymorphism: molecular diagnosis and allele frequencies in Caucasian and African populations. AB - Sulphation, catalysed by members of the sulphotransferase (SULT) enzyme family, is an important component of the body's chemical defence mechanism, but also acts to bioactivate mutagens such as hydroxylated aryl and heterocyclic amines. A major human sulphotransferase, SULT1A1 (P-PST), metabolizes and/or bioactivates many drugs, iodothyronines and hydroxylated aromatic amines. The enzyme activity varies widely within the population and is under genetic control. We have developed an assay detecting a G-->A transition in SULT1A1 that causes an Arg213- >His substitution associated with low SULT activity and altered enzyme properties, and have used it to assess the SULT1A1 genotype in Caucasian (n=293) and African (Nigerian, n=52) populations. We show that the mutant SULT1A1*2 allele is present at frequencies of 0.321 and 0.269 in the Caucasian and African populations respectively. We also demonstrate a significant age-related difference in SULT1A1 genotype within our Caucasian population, with increasing incidence of SULT1A1*1 homozygosity and decreasing incidence of SULT1A1*2 homozygosity with increasing age, indicating a potential association of SULT1A1*1 allozyme(s) with protection against cell and/or tissue damage during aging. PMID- 9854024 TI - Insulin-sensitive regulation of glucose transport and GLUT4 translocation in skeletal muscle of GLUT1 transgenic mice. AB - Skeletal muscle glucose transport was examined in transgenic mice overexpressing the glucose transporter GLUT1 using both the isolated incubated-muscle preparation and the hind-limb perfusion technique. In the absence of insulin, 2 deoxy-d-glucose uptake was increased approximately 3-8-fold in isolated fast twitch muscles of GLUT1 transgenic mice compared with non-transgenic siblings. Similarly, basal glucose transport activity was increased approximately 4-14-fold in perfused fast-twitch muscles of transgenic mice. In non-transgenic mice insulin accelerated glucose transport activity approximately 2-3-fold in isolated muscles and to a much greater extent ( approximately 7-20-fold) in perfused hind limb preparations. The observed effect of insulin on glucose transport in transgenic muscle was similarly dependent upon the technique used for measurement, as insulin had no effect on isolated fast-twitch muscle from transgenic mice, but significantly enhanced glucose transport in perfused fast twitch muscle from transgenic mice to approximately 50-75% of the magnitude of the increase observed in non-transgenic mice. Cell-surface glucose transporter content was assessed via 2-N-4-(l-azi-2,2,2-trifluoroethyl)benzoyl-1,3-bis-(d mannos-4-yloxy)-2-propylamine photolabelling methodology in both isolated and perfused extensor digitorum longus (EDL). Cell-surface GLUT1 was enhanced by as much as 70-fold in both isolated and perfused EDL of transgenic mice. Insulin did not alter cell-surface GLUT1 in either transgenic or non-transgenic mice. Basal levels of cell-surface GLUT4, measured in either isolated or perfused EDL, were similar in transgenic and non-transgenic mice. Interestingly, insulin enhanced cell-surface GLUT4 approximately 2-fold in isolated EDL and approximately 6-fold in perfused EDL of both transgenic and non-transgenic mice. In summary, these results reveal differences between isolated muscle and perfused hind-limb techniques, with the latter method showing a more robust responsiveness to insulin. Furthermore, the results demonstrate that muscle overexpressing GLUT1 has normal insulin-induced GLUT4 translocation and the ability to augment glucose transport activity above the elevated basal rates. PMID- 9854025 TI - Identification of a signal transducer and activator of transcription (STAT) binding site in the mouse metallothionein-I promoter involved in interleukin-6 induced gene expression. AB - Mechanisms of regulation of mouse metallothionein (MT)-I gene expression in response to bacterial endotoxin-lipopolysaccharide (LPS) were examined. Northern blot analysis of hepatic MT-I mRNA in interleukin (IL)-6 or tumour necrosis factor (TNF)-receptor type I knock-out mice demonstrated that IL-6, not TNF alpha, is of central importance in mediating hepatic MT-I gene expression in vivo after LPS injection. In vivo genomic footprinting of the MT-I promoter demonstrated a rapid increase, after LPS injection, in the protection of several guanine residues in the -250 to -300 bp region of the MT-I promoter. The protected bases were within sequences which resemble binding sites for the signal transducers and activators of transcription (STAT) transcription factor family. Electrophoretic mobility-shift assays using oligonucleotides from footprinted MT I promoter regions showed that injection of LPS resulted in a rapid increase in the specific, high-affinity, in vitro binding of STAT1 and STAT3 to a binding site at -297 bp (TTCTCGTAA). Western blotting of hepatic nuclear proteins showed that the time-course for changes of total nuclear STAT1 and STAT3 after LPS injection paralleled the increased complex formation in vitro using this oligonucleotide, and binding was specifically competed for by a functional STAT binding site from the rat alpha2-macroglobulin promoter. Furthermore, the MT-I promoter -297 bp STAT-binding site conferred IL-6 responsiveness in the context of a minimal promoter in transient transfection assays using HepG2 cells. This study suggests that the effects of LPS on hepatic MT-I gene expression are mediated by IL-6 and involve the activation of STAT-binding to the proximal promoter. PMID- 9854026 TI - The novel sphingosine 1-phosphate receptor AGR16 is coupled via pertussis toxin sensitive and -insensitive G-proteins to multiple signalling pathways. AB - In the present study, we determined the agonist specificity and the signalling mechanisms of a putative sphingosine 1-phosphate (S1P) receptor, AGR16. In CHO cells transiently transfected with an AGR16 expression vector, but not in cells transfected with an empty vector, the addition of a low concentration of S1P (1 nM) caused an increase in the intracellular free Ca2+ concentration ([Ca2+]i) by mobilization of Ca2+ from both intra- and extra-cellular pools. To determine the spectrum of agonists for AGR16, we employed K562 cells, which in the naive state do not respond at all to either S1P or structurally related lipids with an increase in [Ca2+]i. In K562 cells stably expressing AGR16, S1P and sphingosylphosphorylcholine (SPC) dose-dependently increased [Ca2+]i with half maximal values of 3 nM and 100 nM respectively. In CHO cells stably expressing AGR16 (CHO-AGR16), but not in parental CHO cells, we observed specific binding of [32P]S1P, which was displaced by unlabelled S1P and SPC. In CHO-AGR16 cells, but not in parental CHO cells, S1P stimulated the production of inositol phosphates and Ca2+ mobilization which was only 30% inhibited by pertussis toxin (PTX), different from the case of the recently identified S1P receptor EDG1. Also in CHO AGR16 cells, but not in CHO cells, S1P at higher concentrations activated mitogen activated protein kinase (MAPK) in a PTX-sensitive and Ras-dependent manner. S1P also induced the activation of two stress-activated MAPKs, c-Jun N-terminal kinase and p38, in a manner that was totally insensitive to PTX. In CHO-AGR16 cells, S1P induced stress-fibre formation, with an increase in myosin light chain phosphorylation, in a PTX-insensitive and Rho-dependent manner. S1P also induced an increase in the cellular cAMP content in CHO-AGR16 cells, which contrasts sharply with the case of EDG1. These results establish that the S1P receptor AGR16 is coupled via both PTX-sensitive and -insensitive G-proteins to multiple effector pathways. PMID- 9854027 TI - Age-related changes in the synthesis of link protein and aggrecan in human articular cartilage: implications for aggregate stability. AB - The rates of incorporation of radiolabelled leucine into aggrecan and link protein have been measured in human articular cartilage of different ages. Aggrecan and link protein were purified in the A1 fraction of CsCl gradients as a result of their ability to form high-buoyant-density proteoglycan aggregates with hyaluronic acid. Separation of the aggrecan from the link protein was achieved by Mono Q anion-exchange chromatography. The rates of synthesis of both aggrecan and link protein decreased with age. The age-related decrease in synthesis of aggrecan was paralleled by a decrease in the rate of sulphate incorporation into glycosaminoglycan chains. The synthesis of link protein decreased with age to a greater extent than that of aggrecan such that the ratio of the rates of link protein to aggrecan synthesis decreased from 1 in immature cartilage to 0.2 in mature cartilage. The age-related decrease in link protein synthesis is controlled at least in part by transcriptional or post-transcriptional mechanisms, as shown by the accompanying age-related decrease in link-protein mRNA. The absence of any age-related decrease in aggrecan mRNA suggests that the decrease in synthesis of aggrecan core protein is controlled by a translational mechanism. Measurement of the total tissue content of aggrecan and link protein by radioimmunoassay revealed an age-related increase in the accumulation of these matrix proteins, even though their de novo synthesis was decreasing. This illustrates the importance that the regulation of extracellular post translational modification also has in controlling the overall turnover of the cartilage matrix. PMID- 9854028 TI - Changes in polyamine catabolism in HL-60 human promyelogenous leukaemic cells in response to etoposide-induced apoptosis. AB - The topoisomerase II inhibitor etoposide induced apoptosis in HL-60 cells within 4 h of exposure to the drug, as measured by changes in morphology, DNA fragmentation and cytotoxicity assays. Etoposide-induced apoptosis was accompanied by an increase in polyamine efflux from the cells and a decrease in total polyamine content during the first 24 h of exposure to the drug. Although both enzyme activities increased slightly, there were no significant changes in spermidine/spermine N1-acetyltransferase activity or polyamine oxidase activity. After longer exposures (48-72 h), significant induction of spermidine/spermine N1 acetyltransferase activity and loss of polyamine content occurred. These results suggest that polyamine oxidation and the resultant hydrogen peroxide produced may be associated with the initiation of apoptosis, while induction of the acetyltransferase and overall loss of intracellular polyamines may be involved in the final, possibly necrotic, stages of cell death. PMID- 9854029 TI - The role of the C-terminal region in phosphoglycerate mutase. AB - Removal of the C-terminal seven residues from phosphoglycerate mutase from Saccharomyces cerevisiae by limited proteolysis is associated with loss of mutase activity, but no change in phosphatase activity. The presence of the cofactor 2, 3-bisphosphoglycerate, or of the cofactor and substrate 3-phosphoglycerate together, confers protection against proteolysis. The substrate alone offers no protection. Replacement of either or both of the two lysines at the C-terminus by glycines has only limited effects on the kinetic properties of phosphoglycerate mutase, indicating that these residues are unlikely to be involved in crucial electrostatic interactions with the substrate, intermediate or product in the reaction. However, the double-mutant form of the enzyme is more sensitive to proteolysis and is no longer protected against proteolysis by the presence of cofactor. The proteolysed wild-type and two of the mutated forms of the enzyme show a reduced response to 2-phosphoglycollate, which enhances the instability of the phospho form of the native enzyme. The phosphoglycerate mutase from Schizosaccharomyces pombe, which lacks the analogous C-terminal tail, has an inherently lower mutase activity and is also less responsive to stimulation by 2 phosphoglycollate. It is proposed that the C-terminal region of phosphoglycerate mutase helps to maintain the enzyme in its active phosphorylated form and assists in the retention of the bisphosphoglycerate intermediate at the active site. However, its role seems not to be to contribute directly to ligand binding, but rather to exert indirect effects on the transfer of the phospho group between substrate, enzyme, intermediate and product. PMID- 9854030 TI - Dual coupling of the alpha-thrombin receptor to signal-transduction pathways involving phosphatidylinositol and phosphatidylcholine metabolism. AB - Addition of alpha-thrombin to quiescent IIC9 cells results in the activation of lipid-metabolizing enzymes associated with signal-transduction cascades. These enzymes include phosphatidylinositol (PI)-specific phospholipase C (PI-PLC), phosphatidylcholine (PC)-specific phospholipases C and D and phospholipase A2 (PLA2). Whereas the alpha-thrombin receptor has been shown to couple with PI PLCs, it is not clear whether this receptor, or a putative second receptor, couples to one or more of the other phospholipases. In this report we determine whether the cloned receptor couples to all or a subset of these enzymes. We show that (i) an alpha-thrombin-receptor-activating peptide also elicits the above responses and (ii) addition of enterokinase to IIC9 cells, stably transfected with an alpha-thrombin receptor (enterokinase- responsive alpha-thrombin receptor, EKTR) containing an enterokinase cleavage site in place of an alpha thrombin cleavage site, stimulates both PI and PC hydrolysis, including PLA2. Enterokinase also induces mitogenesis in the IIC9s transfected with EKTR. These results indicate that, in addition to initiating a mitogenic signalling cascade, the cloned alpha-thrombin receptor couples to enzymes involved in generating PC derived, as well as PI-derived, second-messenger molecules in IIC9s. Additionally, using the cells transfected with EKTR, we further demonstrate that only activated, i. e. cleaved, receptors are desensitized. PMID- 9854031 TI - N-lobe versus C-lobe complexation of bismuth by human transferrin. AB - Interactions of recombinant N-lobe of human serum transferrin (hTF/2N) with Bi3+, a metal ion widely used in medicine, have been investigated by both UV and NMR spectroscopy. The bicarbonate-independent stability constant for Bi3+ binding (K*) to hTF/2N was determined to be log K* 18.9+/-0.2 in 5 mM bicarbonate/10 mM Hepes buffer at 310 K, pH7.4. The presence of Fe3+ in the C-lobe of intact hTF perturbed Bi3+ binding to the N-lobe, whereas binding of Bi3+ to the C-lobe was unaffected by the presence of Fe3+ in the N-lobe. Reactions of Bi3+ (as bismuth nitrilotriacetate or ranitidine bismuth citrate) with hTF/2N in solutions containing 10 mM bicarbonate induced specific changes to high-field 1H-NMR peaks. The 1H co-ordination shifts induced by Bi3+ were similar to those induced by Fe3+ and Ga3+, suggesting that Bi3+ binding causes similar structural changes to those induced by hTF/2N. 13C-NMR data showed that carbonate binds to hTF/2N concomitantly with Bi3+. PMID- 9854032 TI - Protein kinase C-theta is specifically localized on centrosomes and kinetochores in mitotic cells. AB - In this study we provide evidence that the protein kinase C (PKC)-straight theta isoenzyme is recruited on to the mitotic spindle in dividing murine erythroleukaemia (MEL) cells and associates specifically with centrosome and kinetochore structures. None of the other PKC isoenzymes (-alpha, -delta, epsilon, -mu and -zeta) expressed by MEL cells shows this localization on the mitotic spindle. An identical subcellular distribution of PKC-straight theta is also observed in dividing murine P3 myeloma cells and human LAN-5 neuroblastoma cells, indicating that this PKC isoenzyme interacts with the mitotic apparatus in mammalian cells. In phorbol-ester-treated non-growing MEL cells, a rapid change in the intracellular distribution of PKC-straight theta occurs. Under these conditions, PKC-straight theta is translocated from the nuclear to the cytosolic cell compartment, an event that is accompanied by phosphorylation of the PKC straight theta molecule and is followed by its down-regulation. The recovery of cell growth capacity results in the concomitant reappearance of PKC-straight theta. Furthermore, when MEL cells acquire the differentiated non-growing phenotype, the level of PKC-straight theta is reduced to less than 5%, suggesting that this PKC isoenzyme is no longer required. We propose that, unlike other members of the PKC family, PKC-straight theta may play a role in cell proliferation. PMID- 9854033 TI - Disruption of the utrophin-actin interaction by monoclonal antibodies and prediction of an actin-binding surface of utrophin. AB - Monoclonal antibody (mAb) binding sites in the N-terminal actin-binding domain of utrophin have been identified using phage-displayed peptide libraries, and the mAbs have been used to probe functional regions of utrophin involved in actin binding. mAbs were characterized for their ability to interact with the utrophin actin-binding domain and to affect actin binding to utrophin in sedimentation assays. One of these antibodies was able to inhibit utrophin-F-actin binding and was shown to recognize a predicted helical region at residues 13-22 of utrophin, close to a previously predicted actin-binding site. Two other mAbs which did not affect actin binding recognized predicted loops in the second calponin homology domain of the utrophin actin-binding domain. Using the known three-dimensional structure of the homologous actin-binding domain of fimbrin, these results have enabled us to determine the likely orientation of the utrophin actin-binding domain with respect to the actin filament. PMID- 9854034 TI - Mouse annexin V genomic organization includes an endogenous retrovirus. AB - Mouse annexin V genomic clones were characterized by restriction analysis, Southern blotting and DNA sequencing. The entire gene spans close to 50 kb of the mouse genome and contains 14 exons ranging in size from 31 bp for exon 2 to 482 bp for exon 13 up to the polyadenylation site. Intron sizes range from 111 bp for intron 1b to more than 17 kb for intron 2. Non-coding exon 1 is present in two alternative forms separated by approx. 7.4 kb, and the two promoters associated with exons 1a and 1b are quite distinct. The upstream promoter has a TATA box and may direct the limited, tissue-specific expression of mRNA transcripts containing exon 1a. The downstream, TATA-less promoter has high G+C content, and exon 1b predominates among abundantly expressed mRNA species. The conservation of certain cis-elements, including Sp1, AP2, gamma-IRE and NF-IL6, in orthologous species of annexin V genes points to their possible role in trans-acting protein factor binding and gene regulation. Primer-extension analysis revealed multiple origins for transcription, with principal start sites 100-150 bp upstream of the ATG start codon in exon 2. Intron 4 was longer than that previously identified in the orthologous rat gene due to the integration of an apparently complete copy of the murine endogenous retrovirus element, MuERV-L. Phylogenetic analysis of annexin V from 12 species and the presence of neighbouring loci with paralogous counterparts linked to annexin VI pointed to the common ancestry of these genes via chromosomal duplication more than 600 million years ago. PMID- 9854035 TI - Secretion and apparent activation of human hepatic lipase requires proper oligosaccharide processing in the endoplasmic reticulum. AB - Human hepatic lipase (HL) is a glycoprotein with four N-linked oligosaccharide side chains. The importance of glycosylation for the secretion of catalytically active HL was studied in HepG2 cells by using inhibitors of intracellular trafficking, N-glycosylation and oligosaccharide processing. Secretion of HL was inhibited by carbonyl cyanide m-chlorophenylhydrazone (CCCP), monensin, brefeldin A (BFA), tunicamycin, castanospermine and N-methyldeoxynojirimycin, but not by 1 deoxymannojirimycin. Secretion of alpha1-antitrypsin, an unrelated N glycoprotein, was also inhibited by monensin, BFA and tunicamycin, but not by CCCP, castanospermine or N-methyldeoxynojirimycin. Intracellular HL activity decreased with CCCP, tunicamycin, castanospermine and N-methyldeoxynojirimycin, but increased with monensin and BFA. In the absence of protein synthesis de novo, HL activity secreted into the medium was 7.8+/-2.1-fold higher (mean+/-S.D., n=7) than the simultaneous fall in intracellular HL activity. In cells pretreated with monensin or BFA, this factor decreased to 1.3+/-0.5, indicating that the apparent increase in HL activity had already occurred within these cells. After chromatography on Sepharose-heparin, the specific triacylglycerol hydrolase activity of secreted HL was only 1.7+/-0. 3-fold higher than that of intracellular HL, indicating that the secretion-coupled increase in HL activity is only partly explained by true activation. We conclude that oligosaccharide processing by glucosidases in the endoplasmic reticulum is necessary for the transport of newly synthesized human HL, but not alpha1-antitrypsin, to the Golgi, where the catalytic activity of HL is unmasked. PMID- 9854037 TI - eNOS-overexpressing mice: too much NO makes the blood pressure low. PMID- 9854036 TI - Gene structure, expression and chromosomal localization of murine theta class glutathione transferase mGSTT1-1. AB - We have isolated and characterized a cDNA and partial gene encoding a murine subfamily 1 Theta class glutathione transferase (GST). The cDNA derived from mouse GSTT1 has an open reading frame of 720 bp encoding a peptide of 240 amino acids with a calculated molecular mass of 27356 Da. The encoded protein shares only 51% deduced amino acid sequence identity with mouse GSTT2, but greater than 80% deduced amino acid sequence identity with rat GSTT1 and human GSTT1. Mouse GSTT1-1 was expressed in Escherichia coli as an N-terminal 6x histidine-tagged protein and purified using immobilized-metal affinity chromatography on nickel agarose. The yield of the purified recombinant protein from E. coli cultures was approx. 14 mg/l. Recombinant mouse GSTT1-1 was catalytically active towards 1, 2 epoxy-3-(p-nitrophenoxy)propane, 4-nitrobenzyl chloride and dichloromethane. Low activity towards 1-menaphthyl sulphate and 1-chloro-2,4-dinitrobenzene was detected, whereas mouse GSTT1-1 was inactive towards ethacrynic acid. Recombinant mouse GSTT1-1 exhibited glutathione peroxidase activity towards cumene hydroperoxide and t-butyl hydroperoxide, but was inactive towards a range of secondary lipid-peroxidation products, such as the trans-alk-2-enals and trans,trans-alka-2,4-dienals. Mouse GSTT1 mRNA is most abundant in mouse liver and kidney, with some expression in intestinal mucosa. Mouse GSTT1 mRNA is induced in liver by phenobarbital, but not by butylated hydroxyanisole, beta napthoflavone or isosafrole. The structure of mouse GSTT1 is conserved with that of the subfamily 2 Theta class GST genes mouse GSTT2 and rat GSTT2, comprising five exons interrupted by four introns. The mouse GSTT1 gene was found, by in situ hybridization, to be clustered with mouse GSTT2 on chromosome 10 at bands B5 C1. This region is syntenic with the location of the human Theta class GSTs clustered on chromosome 22q11.2. Similarity searches of a mouse-expressed sequence tag database suggest that there may be two additional members of the Theta class that share 70% and 88% protein sequence identity with mouse GSTT1, but less than 55% sequence identity with mouse GSTT2. PMID- 9854038 TI - Inherited interleukin 12 deficiency in a child with bacille Calmette-Guerin and Salmonella enteritidis disseminated infection. AB - Interferon-gamma receptor ligand-binding chain (IFN-gammaR1) or signaling chain (IFN-gammaR2) deficiency, like interleukin 12 receptor beta1 chain (IL-12Rbeta1) deficiency, predispose to severe infections due to poorly virulent mycobacteria and salmonella. A child with bacille Calmette-Guerin and Salmonella enteritidis infection was investigated. Mutations in the genes for IFN-gammaR1, IFN-gammaR2, IL-12Rbeta1, and other molecules implicated in IL-12- or IFN-gamma-mediated immunity were sought. A large homozygous deletion within the IL-12 p40 subunit gene was found, precluding expression of functional IL-12 p70 cytokine by activated dendritic cells and phagocytes. As a result, IFN-gamma production by lymphocytes was markedly impaired. This is the first discovered human disease resulting from a cytokine gene defect. It suggests that IL-12 is essential to and appears specific for protective immunity to intracellular bacteria such as mycobacteria and salmonella. PMID- 9854039 TI - HIV-1 kills renal tubular epithelial cells in vitro by triggering an apoptotic pathway involving caspase activation and Fas upregulation. AB - HIV-infected patients suffer several renal syndromes, which can progress rapidly from renal insufficiency to end-stage renal disease. Histologically, HIV-induced nephropathy is characterized by prominent tubulopathy with apoptosis of tubular cells. Clinical and experimental evidence suggests that renal injury may be directly related to virus infection. Although HIV-1 is a polytropic and not solely lymphotropic pathogen, the susceptibility of renal cells to HIV-1 remains to be determined. This paper demonstrates in vitro the permissiveness of proximal tubular epithelial cells (PTEC) to HIV-1 and describes the effects of PTEC infection to explain the pathogenesis of tubular damage in vivo. The results indicate that PTEC express HIV-specific receptor and coreceptors and sustain virus replication. We observed that HIV-1 infection causes the death of tubular cells by triggering an apoptotic pathway involving caspase activation. Fas upregulation but not Fas ligand expression was found in the infected PTEC. However, after HIV-1 infection, tubular cells became susceptible to apoptosis induced through Fas stimulation. Caspase inhibition prevented the death of the infected PTEC in spite of persistent viral replication. These findings may explain the prominent histopathology of HIV-associated nephropathy and demonstrate that the apoptosis of nonlymphoid cells can be directly induced by HIV-1. PMID- 9854040 TI - Blunted feedback suppression of SREBP processing by dietary cholesterol in transgenic mice expressing sterol-resistant SCAP(D443N). AB - Feedback regulation of cholesterol biosynthesis is mediated by membrane-bound transcription factors designated sterol regulatory element-binding proteins (SREBP)-1 and -2. In sterol-deprived cultured cells, SREBPs are released from membranes by a proteolytic process that is stimulated by SREBP cleavage activating protein (SCAP), a membrane protein containing a sterol-sensing domain. Sterols suppress SREBP cleavage by blocking the action of SCAP, thereby decreasing cholesterol synthesis. A point mutation in SCAP(D443N) causes resistance to sterol suppression. In this article, we produced transgenic mice that express mutant SCAP(D443N) in liver. In these livers the nuclear content of SREBP-1 and -2 was increased, mRNAs encoding proteins involved in uptake and synthesis of cholesterol and fatty acids were elevated, and the livers were engorged with cholesteryl esters and triglycerides enriched in monounsaturated fatty acids. When the mice were challenged with a high cholesterol diet, cleavage of SREBP-1 and -2 was reduced in wild-type livers and less so in transgenic livers. We conclude that SCAP(D443N) stimulates proteolytic processing of native SREBPs in liver and decreases the normal sterol-mediated feedback regulation of SREBP cleavage, suggesting a central role for SCAP as a sterol sensor in liver. PMID- 9854041 TI - Hypotension and reduced nitric oxide-elicited vasorelaxation in transgenic mice overexpressing endothelial nitric oxide synthase. AB - Nitric oxide (NO), constitutively produced by endothelial nitric oxide synthase (eNOS), plays a major role in the regulation of blood pressure and vascular tone. We generated transgenic mice overexpressing bovine eNOS in the vascular wall using murine preproendothelin-1 promoter. In transgenic lineages with three to eight transgene copies, bovine eNOS-specific mRNA, protein expression in the particulate fractions, and calcium-dependent NOS activity were confirmed by RNase protection assay, immunoblotting, and L-arginine/citrulline conversion. Immunohistochemical studies revealed that eNOS protein was predominantly localized in the endothelial cells of aorta, heart, and lung. Blood pressure was significantly lower in eNOS-overexpressing mice than in control littermates. In the transgenic aorta, basal NO release (estimated by Nomega-nitro-L-arginine induced facilitation of the contraction by prostaglandin F2alpha) and basal cGMP levels (measured by enzyme immunoassay) were significantly increased. In contrast, relaxations of transgenic aorta in response to acetylcholine and sodium nitroprusside were significantly attenuated, and the reduced vascular reactivity was associated with reduced response of cGMP elevation to these agents as compared with control aortas. Thus, our novel mouse model of chronic eNOS overexpression demonstrates that, in addition to the essential role of eNOS in blood pressure regulation, tonic NO release by eNOS in the endothelium induces the reduced vascular reactivity to NO-mediated vasodilators, providing several insights into the pathogenesis of nitrate tolerance. PMID- 9854042 TI - The importance of local mucosal HIV-specific CD8(+) cytotoxic T lymphocytes for resistance to mucosal viral transmission in mice and enhancement of resistance by local administration of IL-12. AB - Although crucial to mucosal vaccine development, the mechanisms of defense against mucosal viral infection are still poorly understood. Protection, cytotoxic T lymphocytes (CTL), and neutralizing antibodies have all been observed, but cause and effect have been difficult to determine. The ability of CTL in the mucosa to mediate protection against mucosal viral transmission has never been proven. Here, we use an HIV peptide immunogen and an HIV-1 gp160 expressing recombinant vaccinia viral intrarectal murine challenge system, in which neutralizing antibodies do not play a role, to demonstrate for the first time that long-lasting immune resistance to mucosal viral transmission can be accomplished by CD8(+) CTL that must be present in the mucosal site of exposure. The resistance is ablated by depleting CD8(+) cells in vivo and requires CTL in the mucosa, whereas systemic (splenic) CTL are shown to be unable to protect against mucosal challenge. Furthermore, the resistance as well as the CTL response can be increased by local mucosal delivery of IL-12 with the vaccine. These results imply that induction of local mucosal CTL may be critical for success of a vaccine against viruses transmitted through a mucosal route, such as HIV. PMID- 9854044 TI - SCID mice containing muscle with human mitochondrial DNA mutations. An animal model for mitochondrial DNA defects. AB - Defects of the mitochondrial genome are important causes of disease. Despite major advances in our investigation of patients, there is no effective therapy. Progress in this area is limited by the absence of any animal models in which we can evaluate treatment. To develop such a model we have injected human myoblasts into the tibialis anterior of SCID mice after inducing necrosis. After injection of normal human myoblasts, regenerating fibers expressed human beta-spectrin, confirming they were derived from fusion of human myoblasts. The stability of the muscle fibers was inferred by demonstrating the formation of motor end plates on the regenerating fibers. In addition, we show the presence of human cytochrome c oxidase subunit II, which is encoded by the mitochondrial genome, in the regenerated fibers. After injection of human myoblasts containing either the A8344G or the T8993C heteroplasmic mitochondrial DNA mutations, human beta spectrin positive fibers were found to contain the mutation at a similar level to the injected myoblasts. These studies highlight the potential value of this model for the study of mitochondrial DNA defects. PMID- 9854043 TI - Identification and characterization of autoreactive T cell responses to bullous pemphigoid antigen 2 in patients and healthy controls. AB - Antibodies against the extracellular domain of bullous pemphigoid antigen 2 (BPAG2) are thought to play a key role in the pathogenesis of bullous pemphigoid (BP), the most frequent autoimmune bullous disease of the skin. Autoreactive T cell responses to BPAG2 were investigated in 16 BP patients and 24 healthy controls by coculture of PBMC with two recombinant BPAG2 proteins (extracellular domain of BPAG2). Primary in vitro T cell responses to BPAG2 were observed in 10/12 BP patients expressing the BP-associated HLA-DQB1*0301 allele and 8/10 DQB1*0301 positive healthy individuals. DQB1*0301 also restricted three autoreactive T cell lines from two BP patients and a healthy donor. In contrast, PBMC from 14 normal patients carrying HLA class II alleles other than DQB1*0301 were not stimulated by BPAG2. Autoreactive BPAG2-specific CD4(+) T cell lines and clones from five BP patients produced both Th1 and Th2 cytokines, whereas three autoreactive T cell lines from three DQB1*0301 positive normal patients produced exclusively IFN-gamma. The absence of BPAG2-specific Th2 cells in healthy individuals strongly suggests that autoreactive Th2 responses to BPAG2 are restricted to BP patients and may thus be critical in the pathogenesis of BP. PMID- 9854045 TI - The development of experimental autoimmune encephalomyelitis in the mouse requires alpha4-integrin but not alpha4beta7-integrin. AB - Because monoclonal antibodies (mAbs) directed against alpha4-integrin and VCAM-1 inhibit the development of experimental autoimmune encephalomyelitis (EAE) in vivo, it has been concluded that the successful therapeutic effect is due to interference with alpha4beta1/VCAM-1-mediated interaction of autoaggressive T cells with the blood-brain barrier. A possible role for alpha4beta7-integrin, or interference with other T cell mediated events during the pathogenesis of EAE, has not been considered. We have compared the effects of mAb therapy on the development of EAE in the SJL/N mouse, using a large panel of mAbs directed against alpha4, beta7, the alpha4beta7-heterodimer, and against VCAM-1. Although encephalitogenic T cells express both alpha4-integrins, mAbs directed against the alpha4beta7-heterodimer or against the beta7-subunit did not interfere with the development of EAE. In contrast, mAbs directed against alpha4 and VCAM-1 inhibited or diminished clinical or histopathological signs of EAE. Our data demonstrate for the first time that alpha4beta7 is not essential for the development of EAE. Furthermore, our in vitro studies suggest that the therapeutic effect of anti-alpha4-treatment of EAE might also be caused by inhibition of antigen-specific T cell proliferation. PMID- 9854046 TI - Bone morphogenetic proteins induce the expression of noggin, which limits their activity in cultured rat osteoblasts. AB - Bone morphogenetic proteins (BMPs) induce the differentiation of cells of the osteoblastic lineage and enhance the function of the osteoblast. Growth factors are regulated by binding proteins, but there is no information about binding proteins for BMPs in skeletal cells. Noggin specifically binds BMPs, but its expression by cells of the osteoblastic lineage has not been reported. We tested for the expression of noggin and its induction by BMP-2 in cultures of osteoblast enriched cells from 22-d-old fetal rat calvariae (Ob cells). BMP-2 caused a time- and dose-dependent increase in noggin mRNA and polypeptide levels, as determined by Northern and Western blot analyses. The effects of BMP-2 on noggin transcripts were dependent on protein, but independent of DNA synthesis. BMP-2 increased the rates of noggin transcription as determined by nuclear run-on assays. BMP-4, BMP 6, and TGF-beta1 increased noggin mRNA in Ob cells, but basic fibroblast growth factor, platelet- derived growth factor BB, and IGF-I did not. Noggin decreased the stimulatory effects of BMPs on DNA and collagen synthesis and alkaline phosphatase activity in Ob cells. In conclusion, BMPs induce noggin transcription in Ob cells, a probable mechanism to limit BMP action in osteoblasts. PMID- 9854047 TI - Evidence for altered synthesis of type II collagen in patients with osteoarthritis. AB - There is evidence to suggest that the synthesis of type II collagen is increased in osteoarthritis (OA). Using an immunoassay, we show that the content of the C propeptide of type II procollagen (CPII), released extracellularly from the newly synthesized molecule, is directly related to the synthesis of this molecule in healthy and osteoarthritic articular cartilages. In OA cartilage, CPII content is often markedly elevated (mean 7.6-fold), particularly in the mid and deep zones, reaching 29.6% of the content in newborn. Synthesis is also directly related to total collagen II content in OA, suggesting its importance in maintaining collagen content and cartilage structure. The release of CPII from cartilage is correlated directly with cartilage content. However, the increase in CPII in OA cartilage is not reflected in serum, where a significant reduction is observed. Together these studies provide evidence for alterations in procollagen II synthesis in vivo in patients with OA. PMID- 9854048 TI - Donor-derived interferon gamma is required for inhibition of acute graft-versus host disease by interleukin 12. AB - We have demonstrated that a single injection of interleukin (IL)-12 on the day of bone marrow transplantation (BMT) inhibits acute graft-versus-host disease (GVHD) in mice. This effect of IL-12 can be diminished by anti-interferon (IFN)-gamma mAb. To determine the mechanism by which IFN-gamma affects IL-12-mediated GVHD protection, we have compared the effect of IL-12 on GVHD in C57BL/6 wild-type (WT) or IFN-gamma gene knockout (GKO) recipients of fully major histocompatibility complex plus minor antigen-mismatched allogeneic BMT from WT or GKO BALB/c mice. Lethal acute GVHD was readily induced in the absence of IFN gamma. IL-12 inhibited GVHD mortality to a similar extent in WT and GKO recipients of WT allogeneic BMT. However, neither WT nor GKO recipients were protected by IL-12 from GVHD induced by GKO allogeneic BMT. Moreover, the effective inhibition of host-reactive donor T cell activation and expansion that is associated with IL-12-mediated GVHD protection was dependent on the ability of BALB/c donors to produce IFN-gamma. These results demonstrate that (a) acute GVHD can be induced in the absence of IFN-gamma, (b) host IFN-gamma does not play a critical role in IL-12-induced GVHD protection, and (c) the protective effect of IL-12 against GVHD is dependent on the ability of the donor to produce IFN-gamma. PMID- 9854049 TI - Food intake in free-feeding and energy-deprived lean rats is mediated by the neuropeptide Y5 receptor. AB - The new neuropeptide Y (NPY) Y5 receptor antagonist CGP 71683A displayed high affinity for the cloned rat NPY Y5 subtype, but > 1, 000-fold lower affinity for the cloned rat NPY Y1, Y2, and Y4 subtypes. In LMTK cells transfected with the human NPY Y5 receptor, CGP 71683A was without intrinsic activity and antagonized NPY-induced Ca2+ transients. CGP 71683A was given intraperitoneally (dose range 1 100 mg/kg) to a series of animal models of high hypothalamic NPY levels. In lean satiated rats CGP 71683A significantly antagonized the increase in food intake induced by intracerebroventricular injection of NPY. In 24-h fasted and streptozotocin diabetic rats CGP 71683A dose-dependently inhibited food intake. During the dark phase, CGP 71683A dose-dependently inhibited food intake in free feeding lean rats without affecting the normal pattern of food intake or inducing taste aversion. In free-feeding lean rats, intraperitoneal administration of CGP 71683A for 28 d inhibited food intake dose-dependently with a maximum reduction observed on days 3 and 4. Despite the return of food intake to control levels, body weight and the peripheral fat mass remained significantly reduced. The data demonstrate that the NPY Y5 receptor subtype plays a role in NPY-induced food intake, but also suggest that, with chronic blockade, counterregulatory mechanisms are induced to restore appetite. PMID- 9854051 TI - Corticosteroid-resistant bronchial asthma is associated with increased c-fos expression in monocytes and T lymphocytes. AB - Unstimulated peripheral blood mononuclear cells (PBMCs) from corticosteroid resistant (CR) but not corticosteroid-sensitive (CS) asthmatics demonstrate increased activating peptide-1 (AP-1)- and decreased glucocorticoid receptor (GR) DNA binding. We test whether these abnormalities are associated with excessive generation of c-fos, the inducible component of AP-1. The c-fos transcription rate, mRNA and protein levels, and GR-DNA binding were quantitated in PBMCs, T cells, and monocytes from CS, CR, and nonasthmatic subjects. There was a 1.7-, 4.2-, and 2.3-fold greater increase in the baseline c-fos transcription rate, mRNA expression, and protein levels, respectively, in PBMCs derived from CR compared with CS patients. At optimal stimulation with PMA, there was a 5.7-, 3.4 , and 2-fold greater increase in the c-fos transcription rate, mRNA accumulation, and protein levels, respectively, in CR compared with CS PBMCs. These abnormalities were detected in both the T cell and monocyte subpopulations. PMA stimulation converted PBMCs from a CS to a CR phenotype and was associated with direct interaction between c-Fos and the GR. Pretreatment of PBMCs from CR patients with c-fos antisense oligonucleotides enhanced GR-DNA binding activity in CR PBMCs stimulated with dexamethasone. We suggest that increased c-fos synthesis provides a major mechanism for the increased AP-1- and decreased GR- DNA binding seen in CR asthma. PMID- 9854050 TI - Host defense molecule polymorphisms influence the risk for immune-mediated complications in chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is an inherited disorder of phagocyte function in which defective superoxide production results in deficient microbicidal activity. CGD patients suffer from recurrent, life-threatening infections, and nearly half develop chronic gastrointestinal (GI) complications (colitis, gastric outlet obstruction, or perirectal abscess) and/or autoimmune/rheumatologic disorders (AIDs). To identify genetic modifiers of disease severity, we studied a cohort of 129 CGD patients, in whom seven candidate genes (myeloperoxidase [MPO], mannose binding lectin [MBL], Fcgamma receptors IIa, IIIa, IIIb, TNF-alpha, and IL-1 receptor antagonist), each containing a physiologically relevant polymorphism predicted to influence the host inflammatory response, were selected for analysis. Genotypes of MPO (P = 0.003) and FcgammaRIIIb (P = 0.007) were strongly associated with an increased risk for GI complications, while an FcgammaRIIa (P = 0.05) genotype was suggestive for an association. Patients with all three associated genotypes had the highest risk for GI complications (P < 0.0001). The risk of AIDs was strongly associated with variant alleles of MBL (P = 0.01) and weakly associated with an FcgammaRIIa genotype (P = 0.04). Patients with variant forms of both MBL and FcgammaRIIa had the highest risk of developing an AID (P = 0.003). PMID- 9854052 TI - 5-oxo-ETE induces pulmonary eosinophilia in an integrin-dependent manner in Brown Norway rats. AB - We have shown previously that the 5-lipoxygenase product 5-oxo-6,8, 11,14 eicosatetraenoic acid (5-oxo-ETE) is a highly potent eosinophil chemoattractant in vitro. To determine whether this substance can induce pulmonary eosinophil infiltration in vivo, it was administered to Brown Norway rats by tracheal insufflation. Eosinophils were then counted in lung sections that had been immunostained with an antibody to eosinophil major basic protein. 5-Oxo-ETE induced a dramatic increase in the numbers of eosinophils (ED50, 2.5 microg) around the walls of the airways, which reached maximal levels (five times control levels) between 15 and 24 h after administration, and then declined. LTB4 also induced pulmonary eosinophil infiltration with a similar ED50 but appeared to be somewhat less effective. In contrast, LTD4 and LTE4 were inactive. 5-Oxo-ETE induced eosinophilia was unaffected by the LTB4 and PAF antagonists LY255283 and WEB 2170, respectively. However, it was inhibited by approximately 75% by monoclonal antibodies to CD49d (VLA-4) or CD11a (LFA-1) but was not significantly affected by an antibody to CD11b (Mac-1). In conclusion, 5-oxo-ETE induces pulmonary eosinophilia in Brown Norway rats, raising the possibility that it may be a physiological mediator of inflammation in asthma. PMID- 9854053 TI - Novel AE1 mutations in recessive distal renal tubular acidosis. Loss-of-function is rescued by glycophorin A. AB - The AE1 gene encodes band 3 Cl-/HCO3- exchangers that are expressed both in the erythrocyte and in the acid-secreting, type A intercalated cells of the kidney. Kidney AE1 contributes to urinary acidification by providing the major exit route for HCO3- across the basolateral membrane. Several AE1 mutations cosegregate with dominantly transmitted nonsyndromic renal tubular acidosis (dRTA). However, the modest degree of in vitro hypofunction exhibited by these dRTA-associated mutations fails to explain the disease phenotype in light of the normal urinary acidification associated with the complete loss-of-function exhibited by AE1 mutations linked to dominant spherocytosis. We report here novel AE1 mutations linked to a recessive syndrome of dRTA and hemolytic anemia in which red cell anion transport is normal. Both affected individuals were triply homozygous for two benign mutations M31T and K56E and for the loss-of-function mutation, G701D. AE1 G701D loss-of-function was accompanied by impaired trafficking to the Xenopus oocyte surface. Coexpression with AE1 G701D of the erythroid AE1 chaperonin, glycophorin A, rescued both AE1-mediated Cl- transport and AE1 surface expression in oocytes. The genetic and functional data both suggest that the homozygous AE1 G701D mutation causes recessively transmitted dRTA in this kindred with apparently normal erythroid anion transport. PMID- 9854054 TI - Rapid identification of nine microorganisms causing acute respiratory tract infections by single-tube multiplex reverse transcription-PCR: feasibility study. AB - Acute respiratory tract infections (ARIs) are leading causes of morbidity and, in developing countries, mortality in children. A multiplex reverse transcription PCR (RT-PCR) assay was developed to allow in one test the detection of nine different microorganisms (enterovirus, influenza A and B viruses, respiratory syncytial virus [RSV], parainfluenzaviruses type 1 and type 3, adenovirus, Mycoplasma pneumoniae, and Chlamydia pneumoniae) that do not usually colonize the respiratory tracts of humans but, if present, must be assumed to be the cause of respiratory disease. Clinical samples from 1,118 children admitted to the Department of Pediatrics because of an ARI between November 1995 and April 1998 were used for a first clinical evaluation. Detection of one of the microorganisms included in the assay was achieved for 395 of 1,118 (35%) clinical samples, of which 37.5% were RSV, 20% were influenza A virus, 12.9% were adenovirus, 10.6% were enterovirus, 8.1% were M. pneumoniae, 4.3% were parainfluenzavirus type 3, 3.5% were parainfluenzavirus type 1, 2.8% were influenza B virus, and 0.2% were C. pneumoniae. Seasonal variations in the rates of detection of the different organisms were observed, as was expected from the literature. The levels of concordance with the data obtained by commercially available enzyme immunoassays were 95% for RSV and 98% for influenza A. The results show that the multiplex RT PCR-enzyme-linked immunosorbent assay is a useful and rapid diagnostic tool for the management of children with ARI. Studies of the overall benefit of this method with regard to the use of antibiotics, the use of diagnostic procedures including additional microbiological tests, and hospitalization rate and duration are warranted. PMID- 9854055 TI - Clinical spectrum of infections due to the newly described Actinomyces species A. turicensis, A. radingae, and A. europaeus. AB - Over a 7-year period, we isolated 294 Actinomyces-like organisms (ALOs) which were not clearly identifiable. Using well-defined probes coding for sequences specific for recently described Actinomyces species (A. turicensis, A. radingae, and A. europaeus), we were able to identify 128 strains. The majority belonged to the A. turicensis species. A. radingae was found only in patients with skin related pathologies. A. europaeus was also detected in patients with urinary tract infections. The main sources of A. turicensis were genital infections, followed by skin-related and urinary tract infections. Additional clinical pictures were appendicitis, cholecystitis, ear, nose, and throat infections, and bacteremia. In a small number of patients these ALOs were found as the only pathogen. Strains of the three species were tested by two widely used biochemical identification methods. A. turicensis was easily identifiable by both these methods. We conclude that these ALOs are not infrequent pathogens and are found in a wide range of human infections. At least A. turicensis is easily identifiable by clinical diagnostic laboratories. PMID- 9854056 TI - Comparison of PCR, culture, and serological tests for diagnosis of Mycoplasma pneumoniae respiratory tract infection in children. AB - For diagnosis of Mycoplasma pneumoniae infection we compared two rapid tests, PCR and the immunoglobulin M immunofluorescence assay (IgM IFA), with culture and the complement fixation test (CFT), in a prospective study among 92 children with respiratory tract infection and 74 controls. Based on positivity of culture and/or CFT as the diagnostic criterion, nine patients (10%) were diagnosed with M. pneumoniae infection. All patients positive by culture were also positive by PCR. In all controls cultures, PCRs, and serological assays were negative, except in one with a positive IgM IFA. The IgM IFA had a low positive predictive value of 50%. Only a combination of PCR (seven patients) and CFT (seven patients) allowed diagnosis of all cases. PMID- 9854057 TI - Disseminated infection due to Chrysosporium zonatum in a patient with chronic granulomatous disease and review of non-Aspergillus fungal infections in patients with this disease. AB - We report the first case of Chrysosporium zonatum infection in a 15-year-old male with chronic granulomatous disease who developed a lobar pneumonia and tibia osteomyelitis while on prophylaxis with gamma interferon. The fungus was isolated from sputum and affected bone, and hyphae were observed in the bone by histopathology. Therapy with amphotericin B eradicated the osteomyelitis and pneumonia, but pneumonia recurred in association with pericarditis and pleuritis during therapy with itraconazole. These manifestations subsided, and no recurrences occurred with liposomal amphotericin B therapy. Infections caused by Chrysosporium species are very rare, and C. zonatum has not previously been reported to cause mycosis in humans. This species, the anamorph of the heterothallic ascomycete Uncinocarpus orissi (family Onygenaceae), is distinguished by its thermotolerance, by colonies which darken from yellowish white to buff, and by club-shaped terminal aleurioconidia borne at the ends of short, typically curved stalks. The case isolate produced fertile ascomata in mating tests with representative isolates. The median (range) MICs for our isolate as well as those for two other human isolates and a nonhuman isolate determined by the National Committee for Clinical Laboratory Standards method adapted for moulds were 128 microg/ml (>128 microg/ml) for flucytosine, and 48 microg/ml (32 to >128 microg/ml) for fluconazole. PMID- 9854058 TI - E test versus agar dilution for antimicrobial susceptibility testing of viridans group streptococci. AB - Viridans group streptococci (VGS) are commonly isolated from the blood of hospitalized patients. The E test represents a convenient method for determining the MICs for VGS, but for this purpose it has not been well validated against reference methods. In this study, 180 unselected VGS isolates were identified to a species level, and the MICs of penicillin, cefuroxime, cefotaxime, and vancomycin were determined by both agar dilution and the E test. Available data regarding demographic and laboratory variables for each VGS bacteremic episode were collected, the significance of each VGS isolate was assessed, and the associations between and among laboratory and clinical variables were investigated. Among all VGS isolates, 68.3% (median of three runs) were found to be fully susceptible to penicillin by agar dilution. The E test and agar dilution showed average agreements (within +/-1 dilution) of 92.2% for penicillin, 95.7% for cefuroxime 91.3% for cefotaxime, and 86.7% for vancomycin. Agreements over serial E tests and serial agar dilutions were excellent for beta-lactam agents (intraclass correlation coefficients, >0.9) but less impressive for vancomycin. Very major error rates for the E test were 0.05). These data demonstrate that the MGIT system is an accurate, nonradiometric alternative to the BACTEC 460TB method for rapid susceptibility testing of M. tuberculosis. PMID- 9854063 TI - Mycobacterium species identification and rifampin resistance testing with high density DNA probe arrays. AB - Species identification within the genus Mycobacterium and subsequent antibiotic susceptibility testing still rely on time-consuming, culture-based methods. Despite the recent development of DNA probes, which greatly reduce assay time, there is a need for a single platform assay capable of answering the multitude of diagnostic questions associated with this genus. We describe the use of a DNA probe array based on two sequence databases: one for the species identification of mycobacteria (82 unique 16S rRNA sequences corresponding to 54 phenotypical species) and the other for detecting Mycobacterium tuberculosis rifampin resistance (rpoB alleles). Species identification or rifampin resistance was determined by hybridizing fluorescently labeled, amplified genetic material generated from bacterial colonies to the array. Seventy mycobacterial isolates from 27 different species and 15 rifampin-resistant M. tuberculosis strains were tested. A total of 26 of 27 species were correctly identified as well as all of the rpoB mutants. This parallel testing format opens new perspectives in terms of patient management for bacterial diseases by allowing a number of genetic tests to be simultaneously run. PMID- 9854064 TI - Klebsiella pneumoniae lipopolysaccharide O typing: revision of prototype strains and O-group distribution among clinical isolates from different sources and countries. AB - We have previously described an inhibition enzyme-linked immunosorbent assay method for the O typing of O1 lipopolysaccharide from Klebsiella pneumoniae which overcomes the technical problems and limitations of the classical O-typing method. In this study, we have extended the method to all of the currently recognized O types. The method was validated by studying the prototype strains that have defined the O groups by the classical tube agglutinatination O-typing method. Based on these results, we confirmed the O types of 60 of 64 typeable strains, and we propose a revised O-antigenic scheme, with minor but necessary changes, consisting of serogroups or serotypes O1, O2, O2ac, O3, O4, O5, O7, O8, and O12. Application of this typing method to 638 K. pneumoniae clinical isolates from Denmark, Spain, and the United States from different sources (blood, urine, and others) showed that up to 80% of these isolates belong to serotypes or serogroups O1, O2, O3, and O5, independently of the source of isolation, and that a major group of nontypeable isolates, representing about 17% of the total, consists of half O+ and half O- strains. Differences were observed, however, in the prevalence of the lipopolysaccharide O types or groups, depending on the country and isolation source. PMID- 9854065 TI - Detection of human immunodeficiency virus type 1 nucleocapsid protein p7 in vitro and in vivo. AB - We developed and evaluated an immunoassay for the detection and quantification of human immunodeficiency virus type 1 (HIV-1) nucleocapsid protein p7 using electrochemiluminescence technology. The assay had a dynamic range of 50 to 20,000 pg/ml and a lower detection limit equivalent to approximately 10(6.5) HIV 1 RNA copies/ml in culture supernatant. In vitro kinetic replication studies showed that the amount of p7 correlated strongly with the amount of p24 (R2 = 0.869; P < 0.0001) and viral RNA (R2 = 0.858; P = 0.0009). On the basis of the p7 and RNA concentrations, we calculated the median p7:RNA ratio to be approximately 1,400 p7 molecules per RNA molecule. HIV-1 p7 could be detected and quantified in culture supernatants of both group M subtype A to E viruses and group O viruses. The presence of p7 in vivo was evaluated in 81 serum samples collected from 62 HIV-1-infected individuals. Five samples were p7 positive, whereas 45 samples were HIV-1 p24 positive. Four of the five p7-positive samples were p24 positive as well. p7 could be detected only when serum HIV-1 RNA levels were greater than 10(6) copies/ml. Anti-p7 antibodies were found in six samples, and all six were p7 negative. In contrast to the in vitro results, it appeared that HIV-1 p7 could not be used as a marker for viral quantification in vivo, since more than 90% of the serum samples were p7 negative. In combination with the low prevalence of anti-p7 antibodies, this may, in turn, be advantageous: the p7 assay may be a good alternative to the p24 assay as the readout system for determination of neutralizing activity against HIV-1 in serum or other fluids containing anti-p24 antibodies. PMID- 9854066 TI - Quantitative detection of hepatitis B virus DNA in two international reference plasma preparations. Eurohep Pathobiology Group. AB - Quantitative detection of hepatitis B virus (HBV) in serum or plasma is of significance for monitoring of therapy and establishment of the prognosis of the disease, as well as for infectivity assessment and quality control of the diagnosis. Unfortunately, various commercially available test kits for HBV DNA yielded conflicting quantitative results, with differences of up to a factor of 120. The Eurohep Pathobiology Group has established two reference samples of plasma from HBV carriers and determined as accurately as possible the number of HBV DNA molecules in these samples. Plasma donations from two single highly viremic carriers of HBV genotype A (HBV surface antigen subtype adw2) and genotype D (ayw2/3), respectively, were collected, and coded dilutions of these samples were analyzed by members of the Eurohep Pathobiology Group. Quantitative results from the seven laboratories reporting consistent results were initially divergent. Limiting dilution and nested PCR assays suffered from incomplete DNA extraction. Hybridization assays used inaccurately quantitated cloned DNA as a reference. Two hybridization assays could not be calibrated directly with cloned HBV DNA, because virion-derived DNA reacted much less efficiently. After identification and elimination of these problems, limiting-dilution assays from three laboratories and hybridization assays from two producers generated consistent and concordant results: 2.7 x 10(9) HBV DNA molecules/ml (range, 2.1 x 10(9) to 3.4 x 10(9) HBV DNA molecules/ml) in the plasma from the carrier of genotype A and 2.6 x 10(9) HBV DNA molecules/ml (range, 2.1 x 10(9) to 3.0 x 10(9) HBV DNA molecules/ml in the plasma from the carrier of genotype D. The two Eurohep reference plasma samples have already been used for the standardization of test kits and in quality control trials, and the plasma from the carrier of genotype A will probably be the basis of a World Health Organization reference sample. PMID- 9854067 TI - Multicenter evaluation of the fully automated COBAS AMPLICOR PCR test for detection of Chlamydia trachomatis in urogenital specimens. AB - The fully automated COBAS AMPLICOR CT/NG test for the detection of Chlamydia trachomatis was evaluated in a multicenter trial. Test performance was evaluated for 2,014 endocervical swab and 1,278 urine specimens obtained from women and for 373 urethral swab and 254 urine specimens obtained from men. Culture served as the reference test. Culture-negative, COBAS AMPLICOR-positive specimens that tested positive in a confirmatory PCR test for an alternative target sequence within the C. trachomatis major outer membrane protein gene were resolved as true positives. The overall prevalence of chlamydia was 4.3% in cervical swabs and 11.0% in urethral swabs from men. When the results for each specimen type were considered separately, the resolved sensitivities were 96.5% (83 of 86) for endocervical swab specimens, 95.1% (39 of 41) for urine specimens from women, 100.0% (41 of 41) for urethral swab specimens from men, and 94.4% (17 of 18) for urine specimens from men; the resolved specificities were 99.4% (1,912 of 1,924) for endocervical swab specimens, 99.8% (1,204 of 1,207) for urine specimens from women, 98. 5% (325 of 330) for urethral swab specimens from men, and 100.0% (236 of 236) for urine specimens from men. For the subset of patients from whom both swab and urine specimens were collected, the combined results for both specimen types were used to identify all infected patients. Using these combined reslts as criteria, the resolved sensitivities for the COBAS AMPLICOR test were 82.6% (38 of 46) for endocervical swab specimens, 84.4% (38 of 45) for urine specimens from women, 84.2% (16 of 19) for urethral swab specimens from men, and 89.5% (17 of 19) for urine specimens from men. In comparison, the sensitivity of culture was only 56.5% (26 of 46) for endocervical specimens and 63.2% (12 of 19) for urethral specimens from men. The internal control provided in the COBAS AMPLICOR test revealed that 2.9% of specimens were inhibitory when they were initially tested. Nevertheless, valid results were obtained for 99. 1% of specimens because 68.7% of the inhibitory specimens were not inhibitory when a second aliquot of the original sample was tested. Two additional COBAS AMPLICOR-positive specimens were detected by retesting inhibitory specimens. The COBAS AMPLICOR CT/NG test for the detection of C. trachomatis exhibited equally high sensitivities and specificities with both urogenital swab and urine specimens and, thus, is well suited for use in screening. PMID- 9854068 TI - Molecular epidemiology of outbreaks of gastroenteritis associated with small round-structured viruses in East Anglia, United Kingdom, during the 1996-1997 season. AB - During the winter season from November 1996 to May 1997, 550 fecal specimens were submitted from 94 outbreaks of gastroenteritis occurring in East Anglia, United Kingdom. These specimens were tested for the presence of small round-structured viruses (SRSVs) by electron microscopy, reverse transcriptase PCR, or both methods. SRSVs were shown to be associated with 64 of 94 (68%) of these outbreaks, of which 16 (25%) outbreaks occurred at a single location (Southend) within the region. Twenty-four specimens from 13 of the 16 SRSV-positive outbreaks occurring in Southend were available for genomic analysis, in which divergence within the RNA polymerase region of the SRSV genome was investigated. A further 27 specimens from 17 other SRSV-associated outbreaks, occurring at different locations within East Anglia but at the same time as those at Southend, were also studied. Fifty of the total of 51 (98%) specimens studied were shown to belong to genogroup II, and within this genogroup, 49 of 50 (98%) specimens were shown to be Grimsby-like viruses, with only one Mexico-like strain. Furthermore, phylogenetic analysis of the Grimsby-like viruses indicated clusterings according to the geographical location of the outbreak. One specimen contained a virus belonging to genogroup I, and this had the greatest sequence identity (83%) with Southampton virus. PMID- 9854069 TI - Use of PCR in detection of Mycobacterium avium complex (MAC) bacteremia: sensitivity of the assay and effect of treatment for MAC infection on concentrations of human immunodeficiency virus in plasma. AB - We evaluated the sensitivity and specificity of a PCR-based qualitative test for the rapid diagnosis of Mycobacterium avium-M. intracellulare complex (MAC) bacteremia in patients with AIDS disease. Eleven subjects with newly culture proven MAC bacteremia had the following tests performed at biweekly intervals during the first 8 weeks of therapy: blood culture, Mycobacterium-specific PCR, and quantitative human immunodeficiency virus (HIV) viral-load testing. Mycobacterium genus-specific biotinylated primers were used to amplify a sequence of approximately 582 nucleotides within the 16S rRNA genes of M. avium and M. intracellulare. Detection of the amplified product was performed with an oligonucleotide probe-coated microwell plate combined with an avidin-horseradish peroxidase-tetramethylbenzidine conjugate-substrate system. While not as sensitive as BACTEC culture, PCR detected 17 of 18 specimens which grew >/=40 organisms/ml (94.4% sensitivity) and 9 of 16 specimens which grew /=1/3,200 and 16-fold-lower neutralizing titers to NY-1 than to SN. Reference sera to Hantaan, Seoul, and Prospect Hill viruses also failed to neutralize NY-1. These results indicate that SN and NY-1 define unique hantavirus serotypes and implicate the presence of additional HPS-associated hantavirus serotypes in the Americas. PMID- 9854076 TI - Detection of Pneumocystis carinii DNA in blood specimens from human immunodeficiency virus-infected patients by nested PCR. AB - The detection of Pneumocystis carinii DNA in blood by PCR could be useful for studying the natural history of pneumocystosis and could also be a noninvasive diagnostic method. The results of previous studies are nevertheless conflicting. In our study, we compared three commercially available DNA extraction kits (GeneReleaser, QIAamp Tissue Kit, and ReadyAmp Genomic DNA Purification System) and proteinase K and proteinase K-phenol-chloroform treatments for the extraction of P. carinii DNA from dilutions of a P. carinii f. sp. hominis cyst suspension mixed with human whole blood. A rapid and simple nested PCR protocol which amplifies a portion of the mitochondrial large-subunit rRNA gene was applied to all the extraction products. The QIAmp Tissue Kit was the most effective kit for the isolation of amplification-ready P. carinii DNA and was used with nested PCR for the testing of whole-blood specimens from 35 immunocompetent control patients and 84 human immunodeficiency virus (HIV)-infected patients investigated for pulmonary disease and/or fever. In HIV-infected patients, P. carinii DNA was detected by nested PCR in blood samples from 3 of 14 patients with microscopically proven P. carinii pneumonia, 7 of 22 patients who were considered to be colonized with P. carinii, and 9 of 48 patients who were neither infected nor colonized with P. carinii. P. carinii DNA was not detected in blood specimens from the 35 immunocompetent patients. P. carinii DNA in blood might represent viable P. carinii organisms or DNA complexes released from pulmonary phagocytes. In conclusion, P. carinii DNA may be detected in whole blood from HIV-infected patients, but the nature and the meaning of the circulating form of P. carinii remain to be established. PMID- 9854077 TI - Quantitative analysis of Epstein-Barr virus load by using a real-time PCR assay. AB - To measure the virus load in patients with symptomatic Epstein-Barr virus (EBV) infections, we used a real-time PCR assay to quantify the amount of EBV DNA in blood. The real-time PCR assay could detect from 2 to over 10(7) copies of EBV DNA with a wide linear range. We estimated the virus load in peripheral blood mononuclear cells (PBMNC) from patients with symptomatic EBV infections. The mean EBV-DNA copy number in the PBMNC was 10(3.7) copies/microg of DNA in patients with EBV-related lymphoproliferative disorders, 10(4.1) copies/microg of DNA in patients with chronic active EBV infections, and 10(2.2) copies/microg of DNA in patients with infectious mononucleosis. These numbers were significantly larger than those in either posttransplant patients or immunocompetent control patients without EBV-related diseases. In a patient with infectious mononucleosis, the virus load decreased as the symptoms resolved. The copy number of EBV DNA in PBMNC from symptomatic EBV infections was correlated with the EBV-positive cell number determined by the in situ hybridization assay (r = 0.842; P < 0.0001). These results indicate that the real-time PCR assay is useful for diagnosing symptomatic EBV infection and for monitoring the virus load. PMID- 9854078 TI - Performance characteristics of the BDProbeTec system for direct detection of Mycobacterium tuberculosis complex in respiratory specimens. AB - Strand displacement amplification (SDA) technology has been established in a fully automated system known as BDProbeTec. Target sequences of the insertion sequence IS6110 and the 16S rRNA gene are simultaneously amplified, which thus allows the detection of Mycobacterium tuberculosis complex and, as an additional option, of most Mycobacterium species. Detection occurs via a chemiluminescent microwell assay that employs the simultaneous hybridization and capture of SDA products with a biotinylated capture probe and an alkaline phosphatase detector probe. We have evaluated the performance of the BDProbeTec system in detecting M. tuberculosis complex by testing 799 respiratory specimens and comparing the results to those obtained by conventional diagnostic techniques, i.e. , microscopy and culture (solid and radiometric media). M. tuberculosis was cultivated from 41 specimens, of which 28 (68.4%) were smear positive and 13 (31.6%) were smear negative. The overall sensitivity of the SDA assay was 97.6% (for smear-positive specimens, 100%; for smear-negative specimens, 92.3%), and specificity was 95. 0%. After resolution of the discrepancies by studying the patients' clinical data, sensitivity and specificity were 97.9 and 96.5%, respectively, and positive and negative predictive values were 63.9 and 99.9%, respectively. These preliminary data demonstrate that the BDProbeTec system has promising performance characteristics with respiratory specimens and that it allows the detection of M. tuberculosis complex within hours. PMID- 9854079 TI - Influence of incubation time, inoculum size, and glucose concentrations on spectrophotometric endpoint determinations for amphotericin B, fluconazole, and itraconazole. AB - We addressed the influence of the incubation time (24 h versus 48 h), starting inoculum size (standard inoculum size, approximately 10(3) CFU/ml, versus large inoculum size, approximately 10(4) CFU/ml), and supplementation with 2% glucose of RPMI 1640 medium on the spectrophotometric determination of the MICs of amphotericin B, fluconazole, and itraconazole. We compared the MICs determined spectrophotometrically with those determined by the standard broth macrodilution method (National Committee for Clinical Laboratory Standards approved guideline M27-A). The agreement between the results of the spectrophotometric and standard methods for amphotericin B testing was 100%; this agreement was independent of the inoculum size and incubation time. On the other hand, the agreement for the results for fluconazole testing and itraconazole testing was dependent on the inoculum size and incubation time. With large inoculum size, excellent agreement can be achieved at 24 h. With standard inoculum size, acceptable agreement can be achieved only at 48 h. In contrast to previous observations, the addition of 2% glucose did not have any significant impact on the growth density at 24 h, nor did it improve the agreement with the standard method. Furthermore, supplemental glucose might falsely elevate the MIC at 48 h. PMID- 9854080 TI - Novel intestinal Helicobacter species isolated from cotton-top tamarins (Saguinus oedipus) with chronic colitis. AB - A disease similar to ulcerative colitis in humans has been identified in cotton top tamarins (CTTs) in captivity. The clinical signs include weight loss, diarrhea, and rectal bleeding with the pathological features and biochemical abnormalities of ulcerative colitis. Approximately 25 to 40% of these animals develop colon cancer after 2 to 5 years of captivity. An infectious etiology has been proposed; however, no microbial agent to date has been identified. Helicobacter spp. have been associated with enterocolitis and inflammatory bowel disease (IBD) in humans and animals. Infection with Helicobacter pylori or Helicobacter mustelae is associated with an increased risk of gastric adenocarcinoma and lymphoma of the mucosa-associated lymphoid tissue. Helicobacter hepaticus causes hepatitis, hepatic adenomas, and hepatocellular carcinomas in susceptible strains of mice. The aim of this study was to assess a colony of CTTs with a high incidence of IBD and colon cancer for the presence of colonic Helicobacter spp. A fusiform, gram-negative bacterium with bipolar flagella and periplasmic fibers was isolated from the feces of CTTs. The bacterium grew under microaerobic conditions at 37 and 42 degrees C but not at 25 degrees C, did not hydrolyze urea, was positive for catalase and oxidase, did not reduce nitrate to nitrite, did not hydrolyze indoxyl acetate or alkaline phosphatase, and was resistant to nalidixic acid, cephalothin, and trimethoprim sulfamethoxazole. On the basis of 16S rRNA gene sequence analysis, the organism was classified as a novel Helicobacter species. This is the first Helicobacter isolated from CTTs. Further studies are needed to elucidate the role of this novel Helicobacter sp. in the pathogenesis of ulcerative colitis and colonic adenocarcinoma in CTTs. PMID- 9854081 TI - Homogeneity of 16S-23S ribosomal intergenic spacer regions of Tropheryma whippelii in Swiss patients with Whipple's disease. AB - The current genetic strategies used to identify Tropheryma whippelii, the putative agent of Whipple's disease, are based on PCR-mediated amplification of a part of its 16S rRNA gene (16S rDNA). Because there is very little intraspecies variation in these molecules, they are not suitable as targets for epidemiologic investigations. However, the intergenic spacer region between the 16S and 23S rDNAs is usually much more variable and has repeatedly been used for epidemiologic purposes. We have therefore amplified the spacer region of T. whippelii directly from clinical specimens from nine independent Swiss patients with Whipple's disease by PCR with primers complementary to the 3' and 5' ends of the 16S and 23S rDNAs, respectively. The amplicons were directly sequenced and the sequences were compared to the T. whippelii reference sequence in GenBank/EMBL (accession no. X99636). Complete sequence homogeneity was found between the samples from our nine patients; the spacer sequence was also identical to the reference sequence. However, the sequences corresponding to the 3' and 5' ends of the 16S and the 23S rDNAs of T. whippelii, respectively, differed from the respective sequences in GenBank/EMBL. The same sequence found in our patients was then found in a sample from the German patient from which the published sequence had been derived. We conclude that the 16S-23S rDNA spacer region seems to be very conserved in T. whippelii and that the respective reference entry in public databases should be revised. PMID- 9854082 TI - Detection of penicillin susceptibility in Streptococcus pneumoniae by pbp2b PCR restriction fragment length polymorphism analysis. AB - A PCR-restriction fragment length polymorphism strategy directed against the pbp2b gene was evaluated for identification of penicillin susceptibility. A total of 106 United Kingdom (U.K.), 30 Danish, and 11 Papua New Guinean strains were tested. Of the U.K. strains, all the susceptible and all but one of the resistant isolates were correctly assigned. By using conventional definitions of "not resistant" and "not susceptible," the sensitivities were 97. 5 and 94.4%, the specificities were 100 and 98.9%, the positive predictive values were 100 and 94.4%, and the negative predictive values were 93.1 and 98.9%, respectively. This technique may allow susceptible (MIC, <0.1 mg/liter) and resistant (MIC, >1 mg/liter) isolates to be distinguished in a single PCR. PMID- 9854083 TI - Nucleic acid-based cross-linking assay for detection and quantification of hepatitis B virus DNA. AB - A nucleic acid photo-cross-linking technology was used to develop a direct assay for the quantification of hepatitis B virus (HBV) DNA levels in serum. Cross linker-modified DNA probes complementary to the viral genomes of the major HBV subtypes were synthesized and used in an assay that could be completed in less than 6 h. The quantification range of the assay, as determined by testing serial dilutions of Eurohep HBV reference standards and cloned HBV DNA, was 5 x 10(5) to 3 x 10(9) molecules of HBV DNA/ml of serum. Within-run and between-run coefficients of variation (CVs) for the assay were 4. 3 and 4.0%, respectively. The assay was used to determine HBV DNA levels in 302 serum samples, and the results were compared to those obtained after testing the same samples with the Chiron branched-DNA (bDNA) assay for HBV DNA. Of the samples tested, 218 were positive for HBV DNA by both assays and 72 gave results below the cutoff for both assays. Of the remaining 12 samples, 10 were positive for HBV DNA by the cross linking assay only; the 2 other samples were positive by the bDNA assay only. Twenty-eight samples had to be retested by the bDNA assay (CV, >20% between the results obtained from the testing of each sample in duplicate), whereas only three samples required retesting by the cross-linking assay. The correlation between the HBV DNA levels, as measured by the two tests, was very high (r = 0.902; P = 0.01). We conclude that the cross-linking assay is a sensitive and reproducible method for the detection and quantification of HBV DNA levels in serum. PMID- 9854084 TI - Rapid identification of up to three Candida species in a single reaction tube by a 5' exonuclease assay using fluorescent DNA probes. AB - We used fungus-specific PCR primers and species-specific DNA probes to detect up to three Candida species in a single reaction tube by exploiting the 5' to 3' exonuclease activity of Taq DNA polymerase. Probes to the internal transcribed spacer region of the rRNA gene were labeled at the 5' end with one of three fluorescent reporter dyes, 6-carboxy-fluorescein (FAM), tetrachloro-6-carboxy fluorescein (TET), or hexachloro-6-carboxy-fluorescein (HEX), and at the 3' end with a quencher dye, 6-carboxy-tetramethyl-rhodamine. During PCR amplification, each reporter dye emits a characteristic wavelength as it is cleaved from its specific target DNA and from the quencher dye. Therefore, signals from up to three probes can be detected simultaneously during the PCR assay. Six probes were designed for use in this study: CA-FAM, CT-TET, and CP-HEX were added to one tube to simultaneously detect the typically fluconazole-sensitive species C. albicans, C. tropicalis, and C. parapsilosis, respectively. CG-FAM and CK-TET were added to a second tube to simultaneously detect the typically more innately fluconazole resistant species C. glabrata and C. krusei, respectively. All-CAN-TET, a Candida genus probe, was added to a third tube to detect DNAs from all Candida species tested. DNAs recovered from 61 blood culture bottles, including 23 positive for C. albicans, 18 positive for C. glabrata, 6 positive for C. tropicalis, 6 positive for C. krusei, 5 positive for C. parapsilosis, and 3 positive for mixed fungemias, were tested. Control samples included those from blood culture bottles with no growth (n = 10) or from patients with confirmed bacteremia (n = 10). Probes detected and correctly identified the organisms in 58 of 61 specimens (95.1%) and gave no false-positive results. This method is simple and rapid and does not require post-PCR hybridization and incubation steps. It is sensitive and specific for the detection and identification of Candida species from blood culture bottles, including those containing mixtures of Candida species, and should facilitate an earlier specific diagnosis, leading to more appropriately targeted antifungal drug therapy. PMID- 9854085 TI - Proficiencies of three anaerobic culture systems for recovering periodontal pathogenic bacteria. AB - Anaerobic culture is employed routinely in the primary isolation of periodontal pathogenic bacteria. However, little or no data exist on the relative abilities of the Coy anaerobic chamber (Coy Laboratory Products, Grass Lake, Mich.), the GasPak (Becton Dickinson Microbiology Systems, Cockeysville, Md.), and the AnaeroPack (Mitsubishi Gas Chemical America, Inc., New York, N.Y.) systems to grow important periodontal species, including Porphyromonas gingivalis, Prevotella intermedia/nigrescens, Bacteroides forsythus, Eubacterium species, Campylobacter species, Fusobacterium species, and Peptostreptococcus micros. A total of 78 specimens from advanced periodontitis lesions were collected anaerobically, plated on enriched blood agar medium, and incubated at 35 degrees C for 5 to 7 days in each anaerobic culture system. The three culture systems were equally efficient in isolating Porphyromonas gingivalis and Prevotella intermedia/nigrescens. The Coy anaerobic chamber yielded the highest proportional recoveries of Campylobacter (P = 0.0001; nonparametric analysis of variance) and Eubacterium (P = 0.009). The Coy anaerobic chamber and the GasPak system demonstrated higher proportional recoveries of Bacteroides forsythus (P = 0.0006) and Peptostreptococcus micros (P = 0.0001) than the AnaeroPack system. The AnaeroPack system was most efficient in growing Fusobacterium species (P = 0.0001). Overall, the Coy anaerobic chamber and the GasPak system showed the highest proportional recoveries of putative periodontal pathogens, but the recoveries by the various anaerobic test systems varied considerably from sample to sample. PMID- 9854086 TI - False-positive Gen-Probe direct Mycobacterium tuberculosis amplification test results for patients with pulmonary M. kansasii and M. avium infections. AB - The Gen-Probe Amplified Mycobacterium Tuberculosis Direct (MTD) test has been approved for use in the United States for the rapid diagnosis of pulmonary tuberculosis in patients with acid-fast smear-positive sputum samples since 1996. Four patients infected with human immunodeficiency virus and one chronic pulmonary-disease patient seen in our institutions with abnormal chest radiographs and fluorochrome stain-positive sputa were evaluated for tuberculosis, including performance of the MTD test on expectorated sputum samples. Three of these five patients' sputa were highly smear-positive (i.e. , more than 100 bacilli per high-power field), while two patient's sputa contained 1 to 10 bacilli per field. MTD results on sputum specimens from these patients ranged from 43,498 to 193,858 relative light units (RLU). Gen-Probe has defined values of at least 30,000 RLU as indicative of a positive test, i.e., the presence of Mycobacterium tuberculosis RNA. Four of the patients' sputum cultures yielded growth of M. kansasii within 6 to 12 days, and the fifth produced growth of M. avium only. One patient's culture contained both M. kansasii and M. avium, but none of the initial or follow-up cultures from these five patients revealed M. tuberculosis. However, subsequent cultures from three of the patients again revealed M. kansasii. During the period of this study, in which MTD tests were performed on smear-positive sputum specimens from 82 patients, four of seven patients with culture-proven M. kansasii pulmonary infections yielded one or more false-positive MTD tests. The MTD sensitivity observed in this study was 93.8%, and the specificity was 85.3%. Five cultures of M. kansasii (including three of these patients' isolates and M. kansasii ATCC 12478), and cultures of several other species were examined at densities of 10(5) to 10(7) viable CFU/ml by the MTD test. All five isolates of M. kansasii and three of three isolates of M. simiae yielded false-positive test results, with readings of 75,191 to 335,591 RLU. These findings indicate that low-level false-positive MTD results can occur due to the presence of M. kansasii, M. avium, and possibly other Mycobacterium species other than M. tuberculosis in sputum. Low-level positive MTD results of 30,000 to 500,000 RLU should be interpreted in light of these findings. It remains to be determined if the enhanced MTD test (MTD 2) recently released by Gen-Probe will provide greater specificity than that observed in this report with its first-generation test. PMID- 9854087 TI - Molecular fine-specificity analysis of antibody responses to human cytomegalovirus and design of novel synthetic-peptide-based serodiagnostic assays. AB - To identify single immunodominant marker proteins which can replace complex virion antigen in serodiagnostic assays, we investigated in detail the molecular fine specificity of antibody responses in different individuals with latent or active human cytomegalovirus (HCMV) infection. An overview of the HCMV proteins recognized by human antibodies was obtained by immunoblotting. For selected immunodominant proteins the epitope fine specificity of the antibody response was determined by a peptide-scanning enzyme-linked immunosorbent assay (ELISA). Epitope clusters were synthesized as combination peptides and were used for further serologic analysis of immunoglobulin M (IgM) and IgG reactivities with panels of sera from different groups of patients in comparison to those with cytomegalovirus (CMV) virion antigen. Several serum samples had significantly higher reactivities with peptides than with the CMV virion antigen. However, individual serum samples occasionally recognized diverse peptide epitopes, stressing the importance of using combinations of peptides in serologic assays. From these studies we were able to define a specific combination of peptides derived from pp52 (UL44) and pp150 (UL32) for the specific and highly sensitive early detection of HCMV IgM, whereas a combination of peptides from pp150 (UL32), gB (UL55), and pp28 (UL99) was selected to give optimal and specific reactivity with HCMV IgG. On the basis of the results obtained with these peptide combinations, new, highly specific serodiagnostic assays were constructed. These assays had sensitivities of 98.9 and 96.4% for IgG and IgM, respectively, in comparison with the results obtained with the "gold standard," the virion antigen based ELISA. From the results of this study we conclude that specific combinations of highly defined synthetic peptides can replace complex HCMV virion extracts used in current serodiagnostics and may add to further standardization of HCMV serology. PMID- 9854089 TI - Metarrhizium anisopliae as a cause of sinusitis in immunocompetent hosts. AB - Metarrhizium anisopliae is a common pathogen of insects and has even been used to control insect populations. It is rarely isolated from human or animal sources, but recently, there have been three reported cases of disease, two in humans and one in a cat. We present our experience with five isolates from human sources, including two that were the apparent causes of two cases of sinusitis in immunocompetent hosts. The first patient was a 36-year-old male with frontal and ethmoid sinusitis, and the second was a 79-year-old female with chronic sinusitis. Both patients underwent surgery, and pathology of the surgical specimens revealed branching hyphae. Cultures grew only Metarrhizium species. Neither patient received antifungal therapy, and both did well postoperatively. The other three isolates were cultured from bronchoalveolar lavage specimens but were not felt to be clinically significant. Antifungal susceptibility testing using the National Committee for Clinical Laboratory Standards macrobroth method revealed that all isolates were resistant to amphotericin B, 5-flucytosine, and fluconazole. Itraconazole and newer azole compounds were more active. Metarrhizium species may cause disease in humans, even those without evidence of immunosuppression, and are apparently highly resistant to amphotericin B in vitro. PMID- 9854088 TI - Healthy cats are commonly colonized with "Helicobacter heilmannii" that is associated with minimal gastritis. AB - Gastric Helicobacter infection in healthy pet cats is not well characterized. We performed endoscopy with gastric biopsy on 15 healthy pet cats that were rigorously screened to exclude underlying or concurrent diseases that might affect Helicobacter colonization. Gastric mucosa biopsy specimens were examined by histology, culture, and PCR for the presence of Helicobacter infection and by histology for the presence of gastritis. Of 15 cats, all but 1 had gastric Helicobacter-like organisms (GHLOs) on examination by light microscopy, and in the one histologically negative cat, GHLOs were detected by PCR. Gastric inflammation was mild or was absent for all cats. No Helicobacter species were identified by culture. Analysis of the 16S rRNA sequence from Helicobacter strains from 10 cats showed that all bacteria were closely related to Helicobacter felis, although there was heterogeneity among the sequences. These results suggest that the gastric mucosa of healthy pet cats is commonly colonized with an uncultivated Helicobacter that is closely related to H. felis, is associated with little or no gastritis, and shows heterogeneity in its 16S rRNA sequence. The epithet "Helicobacter heilmannii" continues to be an appropriate working designation for these bacteria. PMID- 9854090 TI - Usefulness of Leifson staining method in diagnosis of Helicobacter pylori infection. AB - The Leifson staining method was used to diagnose Helicobacter pylori infection and was compared to histology, culture, and the rapid urease test (RUT). Histology gave the best sensitivity (98%), compared to Leifson staining (97%), culture (92%), and RUT (85%) (P < 0.005). Leifson staining is a sensitive, rapid, economical method for diagnosis of H. pylori infection in dyspeptic patients. PMID- 9854091 TI - Rapid identification of Candida glabrata by using a dipstick to detect trehalase generated glucose. AB - Candida glabrata is a yeast frequently isolated from human specimens. Based upon its well-known ability to rapidly hydrolyze trehalose, we have developed a novel and cost-effective test incubating one yeast colony emulsified in 50 microl of citrate buffer (0.1 M [pH 5. 0]) containing 4% (wt/vol) trehalose for 3 h at 37 degrees C. Trehalase-generated glucose is detected with a commercially available dipstick (range, 1.0 to 50 g/liter). For evaluation, consecutive clinical isolates and several reference strains of C. glabrata (n = 160), C. albicans (n = 120), and other yeast species with potential ability for utilization of trehalose (C. dubliniensis, n = 11; C. famata, n = 15; C. guilliermondii, n = 5; C. lusitaniae, n = 16; C. parapsilosis, n = 20; C. tropicalis, n = 34; C. viswanathii, n = 5; Pichia angusta, n = 2; C. zeylanoides, n = 2; Saccharomyces cerevisiae, n = 16; C. neoformans, n = 7) were tested. Identification of C. glabrata is achieved within 3 h, with a specificity of 99.1% and a sensitivity of 98.8% when grown on Sabouraud dextrose agar supplemented with 4% glucose. PMID- 9854092 TI - Comparison of two PCRs for detection of Mycobacterium ulcerans. AB - Two nested PCRs for the detection of Mycobacterium ulcerans were compared by using a collection of 65 clinical specimens. The first method amplifies the gene coding for 16S rRNA, and the second method amplifies a repetitive DNA sequence. The sensitivities of bacterioscopy, culture, 16S rRNA gene PCR, and repetitive sequence PCR were 29, 34, 80, and 85%, respectively. Compared to the 16S rRNA gene PCR, the repetitive-sequence PCR was faster, easier to perform, and less expensive. PMID- 9854094 TI - Identification of Yersinia species by the Vitek GNI card. AB - The Vitek GNI card was used to identify 212 isolates of 10 Yersinia species. Identification was correct for 96.3% of the isolates (156 of 162) to the genus level and for 57.4% of the isolates (93 of 162) to the species level for Yersinia spp. listed in the Vitek database. We recommend additional identification methods for isolates assigned to the genus Yersinia by the Vitek system. PMID- 9854093 TI - Identity of ehrlichial DNA sequences derived from Ixodes ricinus ticks with those obtained from patients with human granulocytic ehrlichiosis in Slovenia. AB - Adult Ixodes ricinus (Acari: Ixodidae) ticks collected near Ljubljana, Slovenia, were tested for the agent of human granulocytic ehrlichiosis (HGE) by using PCR assays based on the 16S rRNA gene. Three (3.2%) of 93 ticks were found to contain granulocytic ehrlichiae. Nucleotide sequences of portions of the bacterial groESL heat shock operon amplified from these ticks were identical or nearly (99.8%) identical to those previously determined for human patients with HGE from Slovenia, providing additional evidence that the ticks were infected with the HGE agent. This study identified I. ricinus as the likely vector for these ehrlichial pathogens of humans in this part of Europe. PMID- 9854095 TI - Determination of trimethoprim-sulfamethoxazole resistance in Streptococcus pneumoniae by using the E test with Mueller-Hinton agar supplemented with sheep or horse blood may be unreliable. The Pneumococcal Study Group. AB - An international, multicenter study compared trimethoprim-sulfamethoxazole MICs for 743 Streptococcus pneumoniae isolates (107 to 244 isolates per country) by E test, using Mueller-Hinton agar supplemented with 5% defibrinated horse blood or 5% defibrinated sheep blood, with MICs determined by the National Committee for Clinical Laboratory Standards broth microdilution reference method. Agreement within 1 log2 dilution and minor error rates were 69.3 and 15.5%, respectively, on sheep blood-supplemented agar and 76.9 and 13.6%, respectively, with horse blood as the supplement. Significant interlaboratory variability was observed. E test may not be a reliable method for determining the resistance of pneumococci to trimethoprim-sulfamethoxazole. PMID- 9854096 TI - Comparison between PCR and detection of antigen in sera for diagnosis of pulmonary aspergillosis. AB - We evaluated the usefulness of PCR and antigen detection for the diagnosis of pulmonary aspergillosis. Forty-four serum samples from patients with pulmonary aspergillosis (33 with pulmonary aspergilloma, 4 with allergic bronchopulmonary aspergillosis, 4 with invasive pulmonary aspergillosis, and 3 with aspergillus pyothorax) were used in this study. PCR detection of Aspergillus DNA in serum samples was successful in 39 patients. Galactomannan antigen was detected by sandwich enzyme-linked immunosorbent assay in 25 patients and by latex agglutination test in 13 patients. Detection of Aspergillus DNA in serum samples by nested PCR had the highest sensitivity of the three methods tested for the diagnosis of pulmonary aspergillosis. PMID- 9854097 TI - Dissemination of high-level penicillin-, extended-spectrum cephalosporin-, and erythromycin-resistant Streptococcus pneumoniae clones in Taiwan. AB - Sixty-seven clinical isolates of Streptococcus pneumoniae (40 of serotype 23F, 19 of serotype 19F, and 8 of serotype 6B) with decreased susceptibilities to penicillin and erythromycin were characterized by antimicrobial susceptibility patterns; DNA restriction endonuclease cleavage profiles of the penicillin binding protein genes pbp1a, pbp2b, and pbp2x; random amplified polymorphic DNA (RAPD) patterns generated by arbitrarily primed PCR; and chromosomal macrorestriction profiles based on pulsed-field gel electrophoresis. A total of 22 clones (identical or closely related pulsotypes and identical RAPD patterns) were identified; 14 clones of 23F, 6 of 19F, and 2 of 6B. Three 23F clones (26 isolates) and one 19F clone (9 isolates) expressed high-level resistance to penicillin, cefotaxime, and erythromycin (MICs >/= 256 microg/ml). These data strongly suggest that multiple high-level penicillin-, extended-spectrum cephalosporin-, and macrolide-resistant clones of S. pneumoniae have been disseminated in Taiwan. PMID- 9854098 TI - Rapid identification of Burkholderia pseudomallei by latex agglutination based on an exopolysaccharide-specific monoclonal antibody. AB - We recently identified a constitutively expressed exopolysaccharide of Burkholderia pseudomallei which is composed of a unique linear tetrasaccharide repeating unit consisting of three galactose residues and one 3-deoxy-D-manno-2 octulosonic acid residue. In this study we developed a latex agglutination test based on monoclonal antibody 3015, which is specific for this exopolysaccharide, and evaluated this test for rapid identification of B. pseudomallei grown on agar plates. All 74 environmental and clinical B. pseudomallei strains tested, originating from different areas of Southeast Asia, northern Australia, and Africa, showed a strong and specific agglutination. B. pseudomallei-like organisms and a variety of other bacteria did not react. In conclusion this monoclonal antibody-based test is a simple, rapid, and highly specific method for identifying B. pseudomallei culture isolates from different geographic areas. PMID- 9854099 TI - Evaluation of the Abbott LCx Mycobacterium tuberculosis assay for direct detection of Mycobacterium tuberculosis complex in human samples. AB - Seven hundred thirty-seven clinical samples from 460 patients were processed for direct detection of Mycobacterium tuberculosis complex by a semiautomated ligase chain reaction commercial assay, the LCx Mycobacterium tuberculosis Assay (LCx assay) from Abbott Laboratories. Results were compared to those of direct microscopy and standard microbiological culture. Of 26 patients (5.7%) with a culture positive for M. tuberculosis, 22 (84.6%) were found positive by the LCx assay. The sensitivity of the LCx assay was 98% for smear-positive samples and 27% for smear-negative samples. With an overall culture positivity rate for M. tuberculosis of 8.3% (61 of 737 samples) and after resolution of discrepant results according to clinical data, the sensitivity, specificity, and positive and negative predictive values of the LCx assay were 78, 100, 95, and 98%, respectively, compared to 85, 100, 100, and 98%, respectively, for culture and 67, 99, 87, and 97%, respectively, for acid-fast staining. In conclusion, the LCx assay proved satisfactory and appears to be an easy-to-use 1-day test which must be used with standard culture methods but can considerably reduce diagnosis time versus culture. However, its clinical interest appears to be limited in our population with low mycobacterial prevalence because of its cost considering the small gain in sensitivity versus direct microscopy. PMID- 9854100 TI - Definition of false-positive reactions in screening for hepatitis C virus antibodies. AB - The rate of false-positive hepatitis C virus enzyme immunoassay results was determined to be at least 10% among 1,814 reactive serum samples based on (i) negative results in an independent confirmation assay, (ii) negative PCR results, and (iii) no patients developing clinical or biochemical signs of hepatitis during a 1-year follow-up. PMID- 9854101 TI - Viral superinfection in previously unrecognized chronic carriers of hepatitis B virus with superimposed acute fulminant versus nonfulminant hepatitis. AB - The role of viral superinfection in hepatitis B surface antigen carriers with superimposed fulminant (n = 60) versus nonfulminant (n = 90) acute hepatitis was studied. The frequency of hepatitis A virus (HAV) (0 versus 2.2%), HCV (18.3 versus 21.1%), HDV (15.0 versus 7.8%), and HEV (1.7 versus 4.4%) infection showed no significant difference, while simultaneous HCV and HDV infection was significantly more prevalent in the former (8.3 versus 0%). Only 3. 6% of fulminant cases and 3.3% of nonfulminant controls were HGV RNA positive. PMID- 9854102 TI - Molecular evidence for transmission of human T-lymphotropic virus type II infection by a human bite. AB - Investigation of a human T-lymphotropic virus type II (HTLV-II) infection in a female Australian blood donor identified a human bite as the likely mode of transmission, confirmed by nucleotide sequencing of the proviral tax/rex from both donor and contact. We believe this to be the first report of the transmission of an HTLV by a human bite. PMID- 9854104 TI - DNA fingerprinting of single colonies of Helicobacter pylori from gastric cancer patients suggests infection with a single predominant strain. AB - In each of six gastric cancer patients, repetitive extragenic palindromic PCR DNA fingerprints of 18 single colonies of Helicobacter pylori from the gastric antrum, corpus, and cardia were identical and matched that of the parental isolate. In three additional gastric cancer patients, 17 of 18 single-colony DNA fingerprints were identical to each other and to the DNA fingerprint of the corresponding parental isolate. PMID- 9854103 TI - Ochrobactrum intermedium infection after liver transplantation. AB - A case of bacteremia due to Ochrobactrum intermedium, with concomitant liver abscesses, in an orthotopic liver transplant recipient is presented. Identical microorganisms were isolated from fecal specimens and from an aspirate of a liver abscess that was indicative of invasion of the graft by gastrointestinal spread. 16S DNA sequence analysis of the blood isolate revealed the recovery of the recently proposed new species O. intermedium, closely related to Ochrobactrum anthropi and Brucella spp. PMID- 9854105 TI - Systemic infection of an immunocompromised patient with Methylobacterium zatmanii. AB - We describe the identification of Methylobacterium zatmanii as the causative agent of bacteremia and fever in an immunocompromised patient. The patient, a 60 year-old man, had a 5-month history of acute myeloid leukemia and had been on chemotherapy throughout this period. Seven days after the onset of neutropenia, the patient developed fever. The combination of ciprofloxacin, co-trimoxazole, imipenem, amikacin, and vancomycin led to a complete defervescence. On subculture from six positive blood cultures, the organism grew only on buffered charcoal yeast extract agar and not on standard agars. Identification by universal PCR and subsequent sequence analysis of the amplified 16S rRNA gene segment was achieved. This identification by molecular biology techniques was confirmed by conventional biochemical tests. To our knowledge, this is the first description of M. zatmanii isolated from patient material. PMID- 9854106 TI - Analysis of Legionella pneumophila serogroup 1 isolates by pulsed-field gel electrophoresis. AB - At a hospital in Halifax, Nova Scotia, Canada, three strains of Legionella pneumophila were detectable based on plasmid content, while the isolates collected at another hospital in Halifax had no plasmids. Genomic DNA was digested with BssHII, SalI, and SpeI and subjected to pulsed-field gel electrophoresis (PFGE). We found no relationship between plasmid profile and PFGE pattern. PMID- 9854107 TI - Evaluation of the oxoid dryspot streptococcal grouping kit for grouping beta hemolytic streptococci. AB - The latex agglutination kits that are widely used for grouping of beta-hemolytic streptococci in clinical laboratories use liquid latex suspensions. The Oxoid Dryspot kit (Oxoid Ltd., Basingstoke, Hampshire, United Kingdom) uses predispensed latex dried onto reaction cards or cardboard strips. All streptococci of groups A (85 strains), B (87 strains), C (30 strains), D (38 strains), F (23 strains), and G (65 strains) were correctly grouped by using these reagents. The Oxoid Dryspot Streptococcal Grouping kit is a reliable method for grouping of the beta-hemolytic streptococci encountered in clinical laboratories. PMID- 9854108 TI - Case of onychomycosis caused by Microsporum racemosum. AB - We report the case of a Spanish 60-year-old female who presented in 1997 with onychomycosis of the left thumbnail following an injury caused by a fresh fish bone. Microsporum racemosum was repeatedly cultured from nail scrapings, and its identity was confirmed by sequencing the isolate's ITS1/ITS2 and 5.8S rRNA regions. The patient was successfully treated with itraconazole, which was administered for 12 weeks. This represents the first case of onychomycosis due to M. racemosum and the first time that this species has been isolated from a human in Europe. PMID- 9854109 TI - Optimal activation of isopsoralen to prevent amplicon carryover. AB - We compared the efficiencies of activation of the photochemical isopsoralen compound 10 and its resulting amplicon neutralizations under conditions with a UV transilluminator box at room temperature (RT) and a HRI-300 UV photothermal reaction chamber at RT and at 5 degrees C. Our data suggest that use of the HRI 300 reaction chamber at 5 degrees C results in a statistically significantly higher degree of amplicon neutralization. PMID- 9854110 TI - Serotyping of Streptococcus pneumoniae by coagglutination with 12 pooled antisera. AB - We report on the performance of a recently introduced commercial chessboard method using 12 antisera, in comparison with that of the 55-antiserum panel used in determining the serogroups and types (SGTs) of Streptococcus pneumoniae, both of which were carried out by a coagglutination technique. Of a total of 150 strains of S. pneumoniae studied, 135 (90%) belonged to the SGTs represented in the 23-valent pneumococcal vaccine; of these, 130 (96.3%) were identified as the same SGTs by both typing methods. The remaining five strains showed cross reactivity with more than two pools by the chessboard method, but could be assigned to a single SGT by the Quellung test. The 96.3% concordance of the chessboard method suggests it can be adopted for determination of the SGTs of S. pneumoniae in laboratories. PMID- 9854111 TI - Increasing incidence and comparison of nalidixic acid-resistant Salmonella enterica subsp. enterica serotype typhimurium isolates from humans and animals. AB - We determined the resistance to quinolone of 309 Salmonella enterica subsp. enterica serotype Typhimurium strains isolated from humans and animals (cattle, pigs, or poultry) in 1995 or 1996. Nalidixic acid resistance increased from 8.5% in 1995 to 18.6% in 1996. The highest resistance levels correlated with a mutation at Ser-83 (or Asp-82). All strains remained ciprofloxacin susceptible. Human and animal isolates were compared by pulsed-field gel electrophoresis, and the banding patterns of the human isolates most closely matched those of the bovine isolates. PMID- 9854112 TI - Improvement of immunoglobulin M capture immunoassay specificity: toxoplasma antibody detection method as a model. AB - In the Toxoplasma gondii immunoglobulin M (IgM) capture fluorometric enzyme immunoassay used as a model, nonspecific responses due to the binding of human IgM to horseradish peroxidase (HRP) conjugates were observed despite the removal of the Fc portion of the immunoglobulin. This interaction may be mediated through the binding of human IgM to the HRP moiety of the conjugate. Addition of polymerized HRP into the reaction mixture reduced nonspecific signals in the majority of low false-positive serum reactions. Other plausible sites of interaction are conserved epitopes of mouse immunoglobulins presenting antigenic similarities with the allotopes of other species. Fragmentation of mouse antimicrobial IgG to Fab' and selection of proper conjugation procedure improved assay specificity. PMID- 9854114 TI - The Guillain-Barre syndrome and the 1992-1993 and 1993-1994 influenza vaccines. AB - BACKGROUND: The number of reports of influenza-vaccine-associated Guillain-Barre syndrome to the national Vaccine Adverse Event Reporting System increased from 37 in 1992-1993 to 74 in 1993-1994, arousing concern about a possible increase in vaccine-associated risk. METHODS: Patients given a diagnosis of the Guillain Barre syndrome in the 1992-1993 and 1993-1994 influenza-vaccination seasons were identified in the hospital-discharge data bases of four states. Vaccination histories were obtained by telephone interviews during 1995-1996 and were confirmed by the vaccine providers. Disease with an onset within six weeks after vaccination was defined as vaccine-associated. Vaccine coverage in the population was measured through a random-digit-dialing telephone survey. RESULTS: We interviewed 180 of 273 adults with the Guillain-Barre syndrome; 15 declined to participate, and the remaining 78 could not be contacted. The vaccine providers confirmed influenza vaccination in the six weeks before the onset of Guillain Barre syndrome for 19 patients. The relative risk of the Guillain-Barre syndrome associated with vaccination, adjusted for age, sex, and vaccine season, was 1.7 (95 percent confidence interval, 1.0 to 2.8; P=0.04). The adjusted relative risks were 2.0 for the 1992-1993 season (95 percent confidence interval, 1.0 to 4.3) and 1.5 for the 1993-1994 season (95 percent confidence interval, 0.8 to 2.9). In 9 of the 19 vaccine-associated cases, the onset was in the second week after vaccination, all between day 9 and day 12. CONCLUSIONS: There was no increase in the risk of vaccine-associated Guillain-Barre syndrome from 1992-1993 to 1993 1994. For the two seasons combined, the adjusted relative risk of 1.7 suggests slightly more than one additional case of Guillain-Barre syndrome per million persons vaccinated against influenza. PMID- 9854115 TI - Human immunodeficiency virus type 1 in the semen of men receiving highly active antiretroviral therapy. AB - BACKGROUND: Highly active antiretroviral therapy can effectively decrease the levels of human immunodeficiency virus type 1 (HIV-1) virions in peripheral plasma and seminal fluid of infected men. Whether the genital tract of HIV-1 infected men who are receiving highly active antiretroviral therapy and who have no detectable virus in the peripheral plasma harbors replication-competent virus is not known. METHODS: We collected peripheral-blood and semen samples from seven men with HIV-1 infections who were receiving highly active antiretroviral therapy and who had no detectable viral RNA (fewer than 50 copies per milliliter) in plasma and analyzed the samples for cell-associated proviral DNA using a quantitative polymerase-chain-reaction assay. Replication-competent viruses were evaluated by cell-coculture assays. Proviral DNA and replication-competent virus obtained from peripheral-blood and seminal cells were also analyzed by sequencing relevant viral genes. RESULTS: Despite the long-term suppression of HIV-1 RNA in the plasma of the seven men, proviral DNA was detected in seminal cells in four. Replication-competent viruses were recovered from peripheral-blood cells in three men and from the seminal cells in two of these three men. The viruses recovered from the seminal cells had no genotypic mutations suggestive of resistance to antiretroviral drugs and were macrophage-tropic, a feature that is characteristic of HIV-1 strains that are capable of being sexually transmitted. CONCLUSIONS: In HIV-1-infected men who are receiving highly active antiretroviral therapy and who have no detectable levels of viral RNA in plasma the virus may be present in seminal cells and therefore may be capable of being transmitted sexually. PMID- 9854117 TI - First-trimester growth and the risk of low birth weight. AB - BACKGROUND: Previous studies have demonstrated a correlation between first trimester size and birth weight. It is not known, however, whether low birth weight is related to first-trimester growth. We sought to determine whether the risk of low birth weight and birth weight that was low for gestational age is related to the size of the embryo or the fetus in the first trimester. METHODS: From a data base of ultrasound records of more than 30,000 pregnancies, we identified women who had no important medical problems, a normal menstrual history, and a first-trimester ultrasound scan in which the crown-rump length of the embryo or fetus had been measured. We examined the relation between the outcome of 4229 pregnancies and the difference between the measured and the expected crown-rump length in the first trimester, expressed as equivalent days of growth. RESULTS: A first-trimester crown-rump length that was two to six days smaller than expected was associated with an increased risk (as compared with a normal or slightly larger than expected crown-rump length) of a birth weight below 2500 g (relative risk, 1.8; 95 percent confidence interval, 1.3 to 2.4), a birth weight below 2500 g at term (relative risk, 2.3; 95 percent confidence interval, 1.4 to 3.8), a birth weight below the fifth percentile for gestational age (relative risk, 3.0; 95 percent confidence interval, 2.0 to 4.4), and delivery between 24 and 32 weeks of gestation (relative risk, 2.1; 95 percent confidence interval, 1.1 to 4.0), but not with delivery between 33 and 36 weeks (relative risk, 1.0; 95 percent confidence interval, 0.7 to 1.5). CONCLUSIONS: Suboptimal first-trimester growth may be associated with low birth weight, low birth-weight percentile, and premature delivery. PMID- 9854116 TI - A dose-dependent increase in mortality with vesnarinone among patients with severe heart failure. Vesnarinone Trial Investigators. AB - BACKGROUND: Vesnarinone, an inotropic drug, was shown in a short-term placebo controlled trial to improve survival markedly in patients with severe heart failure when given at a dose of 60 mg per day, but there was a trend toward an adverse effect on survival when the dose was 120 mg per day. In a longer-term study, we evaluated the effects of daily doses of 60 mg or 30 mg of vesnarinone, as compared with placebo, on mortality and morbidity. METHODS: We enrolled 3833 patients who had symptoms of New York Heart Association class III or IV heart failure and a left ventricular ejection fraction of 30 percent or less despite optimal treatment. The mean follow-up was 286 days. RESULTS: There were significantly fewer deaths in the placebo group (242 deaths, or 18.9 percent) than in the 60-mg vesnarinone group (292 deaths, or 22.9 percent) and longer survival (P=0.02). The increase in mortality with vesnarinone was attributed to an increase in sudden death, presumed to be due to arrhythmia. The quality of life had improved significantly more in the 60-mg vesnarinone group than in the placebo group at 8 weeks (P<0.001) and 16 weeks (P=0.003) after randomization. Trends in mortality and in measures of the quality of life in the 30-mg vesnarinone group were similar to those in the 60-mg group but not significantly different from those in the placebo group. Agranulocytosis occurred in 1.2 percent of the patients given 60 mg of vesnarinone per day and 0.2 percent of those given 30 mg of vesnarinone. CONCLUSIONS: Vesnarinone is associated with a dose-dependent increase in mortality among patients with severe heart failure, an increase that is probably related to an increase in deaths due to arrhythmia. A short-term benefit in terms of the quality of life raises issues about the appropriate therapeutic goal in treating heart failure. PMID- 9854118 TI - Familial gestational hyperthyroidism caused by a mutant thyrotropin receptor hypersensitive to human chorionic gonadotropin. PMID- 9854119 TI - Images in clinical medicine. Capnocytophaga canimorsus sepsis. PMID- 9854120 TI - Primary aldosteronism. PMID- 9854122 TI - Influenza vaccination and the Guillain-Barre syndrome. PMID- 9854124 TI - Inotropic therapy for heart failure. PMID- 9854123 TI - Potential for the transmission of HIV-1 despite highly active antiretroviral therapy. PMID- 9854125 TI - Making medicines safer--the need for an independent drug safety board. PMID- 9854126 TI - Preface PMID- 9854127 TI - Historical aspects of laparoscopic hernia repair. AB - The history of the surgical treatment of groin hernias is reviewed with emphasis on the events leading to the introduction and development of the management of these hernias by a laparoscopic approach. The reasons for a shift from an inguinal to an abdominal approach are presented, and the early results of the latter are considered together with the possible advantages and disadvantages. Both methods rely on the use of small or large pieces of mesh that replace the previous bulwark of hernia repairs, namely the rearrangement of tissue layers. Currently, the challenge of the laparoscopic approach is being met by those espousing the inguinal approach. PMID- 9854128 TI - Totally extraperitoneal laparoscopic hernia repair. AB - Totally extraperitoneal laparoscopic hernia repair is the endoscopic equivalent of the open preperitoneal prosthetic approach. Results from a randomized trial confirm that it is less painful, associated with a more rapid return to normal activity, and associated with an insignificant increase in recurrence rates when compared with open tension-free hernia repair. PMID- 9854129 TI - Does the mesh require fixation? AB - As the technique of laparoscopic mesh repair of groin hernia has evolved, the need for mesh fixation has been questioned. Staple fixation has resulted in nerve injury, and some recurrences have been attributed to the use of a mesh that is too small. Drawing on a successful experience from open preperitoneal techniques, laparoscopic surgeons are now reporting results using large mesh without fixation. Early reports show varying success with the technique. Although the problem of nerve damage has been solved, high recurrence rates are reported in which the technique has been used to repair recurrent hernias. Although further studies are required to establish the causes and natural history of mesh migration, initial experience with the technique appears promising. PMID- 9854131 TI - Overview of nonrandomized studies of laparoscopic hernia repair. AB - Laparoscopic techniques have become established in the field of general surgery. Whereas there is general agreement that laparoscopic cholecystectomy has advantages over the open surgical operation, laparoscopic herniorrhaphy or hernioplasty remains controversial. This report looks at a representation of the available literature to date concentrating on nonrandomized studies. PMID- 9854130 TI - The learning curve for laparoscopic hernia repair. AB - As with other laparoscopic procedures, the learning curve for laparoscopic inguinal hernia repair is longer than most surgeons anticipated. This, combined with a lack of familiarity with the preperitoneal anatomy and the simultaneous adoption of open tension-free hernioplasty, has made it difficult for surgeons to sustain enthusiasm for the laparoscopic hernia repair. PMID- 9854132 TI - Overview of randomized trials in laparoscopic inguinal hernia repair. AB - There are more than 20 randomized trials that compare laparoscopic with open hernia repair. Results of these show a longer operating time, less postoperative pain, a faster return to normal activity, and increased costs for laparoscopic over open hernia repair. Recurrence rates are lower for laparoscopic compared with open nonmesh repairs. PMID- 9854133 TI - Economic evaluation of open versus laparoscopic hernia repair: some pragmatic considerations for the measurement of costs. AB - Laparoscopic hernia repair costs more than open repair. This increase in cost largely is because of the use of disposables. Indirect cost benefits of laparoscopic procedure because of a more rapid return to normal activity are different to calculate but may be present for select groups of patients. PMID- 9854134 TI - Other methods of laparoendoscopic hernia repair: mini-hernia--inguinal hernia repair through a 2-cm incision. AB - Mini-hernia repair is an endoscopic modification of the Lichtenstein tension-free repair. Clinical trials are warranted to determine the efficacy of this procedure and any advantages it may have over the original operation. PMID- 9854135 TI - Percutaneous endoscopic external ring hernioplasty using Dundee inguinal canal retractor. AB - The percutaneous endoscopic external ring (PEER) hernioplasty is a novel technique that uses a minimal access approach to the inguinal canal by the anterior route and reproduces the conventional open tension-free mesh repair. The procedure consists of two stages: an open phase through a 2.5-cm incision over the external ring followed by an endoscopic repair of the defect. During the open stage, the spermatic cord is mobilized from the external ring, blunt finger dissection of the inguinal canal is used to create a space beneath the external oblique aponeurosis, and the hernial sac is dissected down to level of the preperitoneal fascia. The second endoscopic stage is performed after insertion of a telescope-mounted retractor through the external ring. The posterior wall of the inguinal canal is visualized and the hernial defect repaired by a tension free mesh repair (Lichtenstein onlay mesh repair or mesh plug repair). The procedure provides an anatomic approach that is familiar to general surgeons. In a pilot study, the authors have established that the PEER hernioplasty is an easily learned, safe, and cost-effective operation that offers the benefits of minimal access surgery in terms of patient recovery and early return to work, and can be performed as a day case under local or regional anesthesia. PMID- 9854136 TI - Subject index PMID- 9854137 TI - The involvement of the ubiquitin system in Alzheimer's disease (review). AB - Alzheimer's disease, the most common form of senile dementia, affects more than 15 million people world-wide and is characterized by a marked deterioration in memory and all cognitive functions, as a result of a progressive degeneration and loss of cortical and limbic neurons. This process is associated with the presence of both the so-called -amyloid deposits and the cellular neurofibrillary tangles composed mainly of paired helical filaments of aberrantly hyperphosphorylated tau protein. The accumulation of ubiquitin in neurofibrillary tangles and senile plaques (both characteristic of the neuronal abnormalities associated with the disease) is postulated to play a role in the repair mechanism related to neuronal regeneration. PMID- 9854138 TI - Novel pathophysiological aspects of macrophage migration inhibitory factor (review). AB - Macrophage migration inhibitory factor (MIF) was the first T-cell-derived soluble lymphokine to be identified. It was originally found to inhibit the migration of macrophages and activate them at inflammatory loci. During the past few years, however, previously unrecognized properties of MIF have been discovered. It also functions, for example, as a pituitary hormone, glucocorticoid-induced immunomodulator and isomerase. We cloned rat MIF cDNA and reported that the nucleotide sequence of the cDNA predicts a protein consisting of 114 amino acids. Northern blot analysis indicated that the MIF mRNA was expressed in a wide variety of organs, including the brain, kidney, and liver. Following this, we demonstrated definitively that MIF was expressed in a variety of cells, suggesting its involvement in various biological events such as wound healing, atopic dermatitis, and, possibly, diabetes/obesity. Furthermore, we elucidated its physicochemical properties, including the tertiary structures of both human and rat MIF. These tertiary structures showed that this protein forms a homotrimer with each monomer consisting of two beta/alpha/beta motifs, thus resembling 5-carboxymethyl-2-hydroxymuconate isomerase and d-dopachrome tautomerase. From the available data on MIF, including ours, it is considered that the protein is associated not only with immune responses but also with cell growth and differentiation during wound repair and carcinogenesis. Thus, MIF could become a major target protein in a variety of pathophysiological states and anti-MIF antibodies and antagonists could be applied therapeutically in the clinical situation for treatment of various diseases. Bearing this in mind, this review discusses the role of MIF, considering its gene and protein structures as well as its pathophysiological functions in various organs and disease states, finally considering perspectives for the future. PMID- 9854139 TI - In vitro expression and in vivo immunogenicity of Plasmodium falciparum pre erythrocytic stage DNA vaccines. AB - DNA vaccine plasmids were constructed that encoded four pre-erythrocytic antigens from the human malaria parasite Plasmodium falciparum: circumsporozoite protein (PfCSP); sporozoite surface protein 2 (PfSSP2); carboxyl terminus of liver stage antigen 1 (PfLSA-1 c-term); and, exported protein 1 (PfExp-1). Antigen expression was evaluated in vitro by immunoblot analysis of tissue culture cells following transient transfection with each plasmid. Clearly detectable levels of expression depended upon, or were markedly enhanced by, fusion of the antigen encoding sequences in-frame with the initiation complex and peptide leader sequence of human tissue plasminogen activator protein. Mice injected with these plasmids produced antigen specific antibody and cytotoxic T lymphocyte responses. However, the magnitudes of the responses were not always predicted by the in vitro expression assay. The results of this study provided the basis for further testing of these plasmids in primates and the formulation of multi-component pre erythrocytic DNA vaccines for efficacy testing in human volunteers. PMID- 9854140 TI - Gene expression during involution of mammary gland (review). AB - After cessation of lactation, the mammary gland undergoes involution, regressing to a state resembling that of a virgin animal. This phase of mammary gland development is characterized by epithelial cell death and tissue remodeling. To understand molecular mechanisms of mammary gland involution, we identified involution-induced clones by differential screening of a mouse mammary gland cDNA library. Several known genes were induced during mammary gland involution: sulfated glycoprotein-2 (SGP-2), WDNM1, lactoferrin, ferritin heavy chain (FHC), lysozyme and osteopontin genes. Involution of the mammary gland is presumed to be mediated by a decrease in serum prolactin level induced by weaning, but may also involve changes in paracrine or autocrine growth factors. Effects of lactogenic hormones and EGF on the expression of the involution-induced genes were examined in mammary epithelial cells. Insulin, dexamethasone, and prolactin did not influence the expression of the FHC, WDNM1 and SGP-2 genes. However, EGF strongly inhibited the expression of WDNM1 and SGP-2 genes. Our recent results are reviewed and discussed. PMID- 9854141 TI - Spontaneous murine lung adenocarcinoma (P07): A new experimental model to study paraneoplastic syndromes of lung cancer. AB - We report the histological and biological behavior characteristics of a lung tumor (P07) that arose spontaneously in a Balb/c mouse. P07 is a moderately to poorly differentiated adenocarcinoma that secretes granulocyte-macrophage colony stimulating factor (GM-CSF) in culture supernatants. This tumor presents some paraneoplastic syndromes, such as leukocytosis, hypercalcemia and cachexia. taken together with the peripheral blood leukocyte (PBL) counts and serum calcium levels during s.c. tumor growth and after surgery, this study suggests that P07 may be a useful experimental model to study the biology of lung cancer and paraneoplastic syndromes. PMID- 9854142 TI - Low levels of serum acylcarnitine in chronic fatigue syndrome and chronic hepatitis type C, but not seen in other diseases. AB - Recently, we found a serum acylcarnitine (ACR) deficiency in Japanese patients with chronic fatigue syndrome (CFS). To clarify whether this ACR abnormality is a characteristic of CFS or not, we also studied the levels of serum carnitine in Swedish subjects. Both serum ACR and free carnitine (FCR) levels in normal healthy subjects were quite different between Japanese (n=131) and Swedish people (n=46) (p<0.001). However, it is confirmed that Swedish patients with CFS (n=57) also had serum ACR deficiency (p<0.001). When we studied the levels of serum ACR and FCR in Japanese patients with various kinds of diseases (CFS, hematological malignancies, chronic pancreatitis, hypertension, diabetes mellitus, chronic hepatitis type C, psychiatric diseases), a significant decrease in the levels of serum ACR was only found in patients with CFS and chronic hepatitis type C (p<0.001). Therefore, we concluded that ACR deficiency in serum might be a characteristic abnormality in only certain types of diseases. PMID- 9854143 TI - Expression of MAGE genes in renal cell carcinoma. AB - We investigated the expression of MAGE genes in 10 renal cell carcinoma (RCC) cell lines, 50 RCC tumor samples and 5 normal kidney samples using reverse transcription-polymerase chain reaction (RT-PCR). MAGE-1, -2, -3 and -4 genes were expressed in 4, 1, 10 and 3 of 10 RCC cell lines, respectively, and 11, 8, 38 and 15 of 50 RCC samples. In contrast, there was no expression of MAGE genes detected in any of the normal kidneys. The incidence of the expression of plural MAGE genes in high stage RCC was significantly higher than that in low stage RCC. An analysis based on clinicopathological factors revealed that MAGE-4 gene was more frequently expressed in clear cell subtype than in granular cell subtype RCCs. Our results suggest that owing to the high incidence of MAGE gene expression in RCC, a large proportion of patients could be suitable candidates for novel immune therapies involving tumor-specific antigens encoded by MAGE genes. PMID- 9854144 TI - Adenoviral transduction efficiency partly correlates with expression levels of integrin alphavbeta5, but not alphavbeta3 in human gastric carcinoma cells. AB - Adenovirus has attracted much attention as a vector for gene therapy. Integrin alphavbeta3 and alphavbeta5 mediate adenovirus internalization into cells. We examined adenoviral transduction efficiency and expression of integrin alphavbeta3 alphavbeta5 in 9 human gastric carcinoma cell lines. The percentage of -gal-positive cells was more than 85% 3 days after infection with recombinant adenovirus carrying the bacterial LacZ gene (AxCALacZ) at a dose of 25 MOI in 7 cell lines and the transduction efficiency was 32 and 42% in HSC-39 and MKN-28 cells, respectively. Adenoviral transduction efficiency did not correlate with the histological cell types. Flow cytometric analysis revealed relatively high expression of integrin alphavbeta5 in MKN-28 and OCUM-2M cells, followed by MKN 1, MKN-7, MKN-45, MKN-74, TMK-1 and KATO-III cells. HSC-39 cells minimally expressed integrin alphavbeta5. On the other hand, integrin alphavbeta3 was expressed only in MKN-1 cells. These results suggest that the adenovirus vector might be useful for gene transduction into human gastric carcinoma cells and the transduction efficiency partly correlates with the expression levels of integrin alphavbeta5 but not integrin alphavbeta3. PMID- 9854145 TI - Reduced metastatic potential and c-myc overexpression of colon adenocarcinoma cells (Colon 26 line) transfected with nm23-R2/rat nucleoside diphosphate kinase alpha isoform. AB - Increased expression of nm23/nucleoside diphosphate kinase (NDP kinase) has been reported to be associated with both reduced metastatic potential in breast carcinoma and tumor progression in colon adenocarcinoma and lung adenocarcinoma. We examined effects of expression of nm23-R2 rat NDP kinase alpha isoform on mouse adenocarcinoma cells (Colon 26 line) and found a significant reduction of metastatic potential along with overexpression of c-myc. We also found that the proliferation rate of the transformed cells was the same as that of the control cells in culture. These results indicate that the cell growth potential in vitro is irrelevant to metastatic potential of the cells in vivo. PMID- 9854146 TI - Ubiquitin and proteasome gene expression is increased in skeletal muscle of slim AIDS patients. AB - Human biopsies obtained from skeletal muscle of cachectic AIDS patients clearly showed an increased expression (in relation to that of healthy subjects) of the genes encoding for the ubiquitin-ATP-dependent proteolytic system. Increases of 120% and 42% were observed for the 2.4 and 1.2 kb ubiquitin transcripts, respectively. The expression of the C8 proteasome subunit was also increased by 60% in the cachectic AIDS patients in relation to the healthy control subjects. It is suggested that the activation of this proteolytic system (possibly via changes in circulating cytokines, such as TNF) may be responsible for the skeletal muscle waste that often accompanies AIDS. PMID- 9854147 TI - Soluble interleukin-2 receptor and soluble CD8 antigen levels in serum from patients with solid tumors. AB - High levels of soluble lymphocyte antigens have been described in a large number of tumors and, particularly, in hematopoietic neoplasms. As previously reported, many antitumor immune responses are IL-2 dependent: clinical observations indicate that a worse survival in advanced tumor patients is related with a decrease of soluble IL-2 levels. A soluble form of CD8 has been described: as found in Hodgkin's disease and acute lymphoblastic leukemia, sCD8 levels have a prognostic value. To explain the significance of these soluble molecules in solid tumors, we a) determinated sIL-2R and sCD8 in 84 patients; b) correlated the expression of p55 chain of IL-2R and CD8 antigen on the cell-surface of peripheral lymphocytes to sIL-2R and sCD8 levels; c) analyzed endogenous IL-2R levels in patients with lung cancer. An increase of sIL-2R was found in 82% of cases, while high levels of sCD8 were observed in 32%; no correlation was observed between sIL-2R and the expression of p55 on the surface of peripheral lymphocytes: IL-2 levels in patients with NSCLC were significatively reduced, when compared to healthy controls, with an inverse relationship between endogenous IL-2 concentration and sIL-2R levels. Whatever may be the physiopathological mechanism of the increase of sIL-2 observed in solid tumors, this rise may contribute to the immunodepression correlated to neoplastic disease. Therefore, higher levels of sIL-2R/IL-2 ratio has a negative biologic prognostic significance. We think that determinating CD8 antigen in the serum can offer a more sensitive and specific measurement of activation of suppressor/cytotoxic T-lymphocytes. PMID- 9854148 TI - Control of vascular smooth muscle cell growth and its implication in atherosclerosis and restenosis (review). AB - Abnormal proliferation of vascular smooth muscle cells (SMCs) is thought to contribute to neointimal thickening during spontaneous atherosclerosis and vessel renarrowing (restenosis) after angioplasty. This review will focus on the mechanisms underlying cell-cycle control in SMCs and its implication in atherosclerosis and restenosis. Therapeutic approaches that targeted specific cell-cycle control genes or growth regulatory molecules which effectively inhibited neointimal lesion formation will also be discussed. PMID- 9854149 TI - Identification of 55 kDa phosphatidylinositol 4-kinase in HaCaT cells: comparison with the epithelial cell line A431. AB - Phosphatidylinositol kinases play a crucial role in signal transduction in many cell types. The 55 kDa isoform of phosphatidylinositol 4-kinases is a key enzyme in the metabolism of phosphoinositides, which work as regulators of cell function itself or as precursors of signal molecules. The experiments with HaCaT cells presented suggest that the phosphatidylinositol 4-kinase activity in this cell line corresponds to the 55 kDa isoform concerning kinetic parameters and specific activity in comparison with the malignant cell line A431. Km (for ATP and phosphatidyl-inositol) and Ki values (for Ca2+ and adenosine) are in good agreement with the parameters described for other cells. The findings support the idea that the 55 kDa phosphatidylinositol 4-kinase represents a key enzyme in the inositide signal transduction pathway. PMID- 9854150 TI - Insulinotropic action of the polyacetate esters of two non-nutrient monosaccharides in normal and diabetic rats. AB - The polyacetate esters of certain non-nutrient monosaccharides, such as L-glucose and 2-deoxy-D-glucose, were recently reported to display positive insulinotropic action and, hence, proposed as possible tools for stimulation of insulin release in non-insulin-dependent diabetes. In the present study, the secretory response to four carbohydrate esters was compared in islets of both normal and hereditary diabetic Goto-Kakizaki rats. Three major findings are documented. First, in islets exposed to the dimethyl ester of succinic acid (10.0 mmol/l), D mannoheptulose hexaacetate (1.7 mmol/l) was found to stimulate insulin release in both normal and diabetic rats. Second, relative to the control value recorded in the sole presence of the succinic acid ester, the increments in insulin output evoked by D-mannoheptulose hexaacetate, alpha-L-glucose pentaacetate and beta-D glucose pentaacetate (all 1.7 mmol/l) were not lower and, on occasion, even higher in diabetic rats than in control animals. Last, the sole exception to such a rule was encountered in islets exposed to beta-L-glucose pentaacetate, in which case the hexose moiety of the ester might mimic the inhibitory effect of alpha-D glucopyranose upon phosphorylase a-catalyzed glycogenolysis in islets from diabetic rats. These findings reinforce the concept that the insulinotropic action of monosaccharide esters is not solely attributable to the catabolism of their carbohydrate moiety but also to a direct effect of the esters themselves upon a yet unidentified receptor system. They also provide further support to the possible use of the esters of non-nutrient monosaccharides as insulinotropic tools in type-2 diabetes. PMID- 9854151 TI - Chimeric anti-TAG72 receptors with immunoglobulin constant Fc domains and gamma or zeta signalling chains. AB - We recently described the generation and expression of a chimeric T cell receptor with specificity for the tumor antigen TAG72 consisting of the single chain antibody (scFv) B72.3-scFv and the gamma chain of the FcepsilonRI receptor. The corresponding chimeric receptor containing the zeta chain of the TCR as signalling unit is not functionally expressed reflecting that the requirements for functional expression of chimeric receptors containing the gamma signalling chain are apparently different compared to those containing the CD3zeta signalling chain of the TCR. We describe a novel set of chimeric anti-TAG72 receptors including in their extracellular moiety the constant immunoglobulin CH2/3 domains that allow stable expression of chimeric gamma as well as zeta receptors. We designed anti-TAG72 receptors that consist of a scFv fragment derived from an anti-TAG72 second generation antibody (CC49) and of the CH2/3 domains of the human IgG and intracellularily either of the zeta or gamma signalling chain. The recombinant CC49-CH2/3-zeta and CC49-CH2/3-gamma DNA, respectively, was transfected into MD45 T cells and expressed under control of the RSV LTR. Both receptors were found on the cell membrane of transfected cells as demonstrated by flow cytometry analysis using an anti-human IgG Fc antibody directed to the CH2/3 immunoglobulin domains of the chimeric receptor. Specific cross-linking of the chimeric zeta as well as the gamma receptor by antigen or anti-human Ig antibodies resulted in specific activation of transfected cells. Our results demonstrate that both the gamma chain and the zeta chain++ containing receptor are stably expressed and convert T cells to specificity for the TAG72 antigen. This receptor design will facilitate efficient generation of genetically modified peripheral T cells and may provide valuable tools for the cellular immunotherapy of TAG72+ tumors. PMID- 9854152 TI - Oral administration of a trace element preparation and zinc inhibit liver metastasis of 3LL-HH murine tumor cells. AB - A trace element preparation (TEP-Beres Drops Plus) was given to liver metastasizing 3LL-HH tumor bearing mice in the presence and after the removal of the primary spleen tumor. In both models when TEP was provided daily and orally in a dose range of 100-5,000 microgram/kg, the number of liver metastasis decreased at an end-point of day 14 and TEP also had inhibitory effect on the primary tumor. The antitumor action proved to be dependent on tumor burden. There was no change in body weight, size distribution of metastases and in the life span of mice. Zn++ could be one of the antimetastatic components among the trace elements. This is the first report on the antimetastatic effect of trace elements in an experimental tumor model. PMID- 9854153 TI - Mutagenicity of microcystin-LR in human RSa cells. AB - Microcystin-LR (MCLR) is a potent cyclic heptapeptidic hepatotoxin produced by bloom-forming cyanobacteria. The mutagenicity induced by this compound in cultured human RSa cells was found by determination of ouabain-resistant (OuaR) mutation, with the highest frequency at the concentration of 15 microgram/ml. Moreover, base substitution mutations at K-ras codon 12 in genomic DNA, assessed by polymerase chain reaction (PCR) and differential dot-blot hybridization using digoxigenin-labeled probes, were detected in RSa cells 6 days after exposure to MCLR (7.5-15 microgram/ml). PMID- 9854154 TI - Comparison of ploidy status of melanoma metastases in different locations. AB - The main aim of this study was to evaluate the DNA content and ploidy status of 29 melanoma metastases in lymph nodes, as well as 10 liver, 12 brain, 13 lung and 2 gastrointestinal metastases. All cases investigated were either suspicious to be aneuploid or clearly aneuploid. This study demonstrates differences of ploidy related parameters between melanoma metastases of different locations. The 5c exceeding rate values were the lowest in lymph node metastases and the highest in melanoma cells in the brain (p<0.05). The rate of cells in S-phase ranged between 12% in gastrointestinal metastases and 23% in liver metastases. The melanoma cells, which formed liver metastases had smaller area, lower mass and bigger shape factors in comparison with melanoma cells in lymph nodes. PMID- 9854156 TI - Cryosurgery in dermatology. AB - The aim of this article is to provide current information on the clinical development of cutaneous cryoreaction and the indications, complications and contraindications of cutaneous cryosurgery. Successful cutaneous cryosurgery requires rapid freezing and slow thawing, minimum tissue temperatures of -25 degrees C to -60 degrees C and, in malignant lesions, repeated freeze-thaw cycles. Frozen tissue reacts with peripheral erythema immediately following thawing, and consequently with oedema, bulla formation, exudation, mummification, and usually heals with a fine atrophic scar within a 4-week period. Cryosurgery is now considered the treatment of choice in hypertrophic scars and keloids, granuloma annulare and capillary haemangioma of the newborn. It also represents a valuable alternative therapy for various skin diseases, including common warts, solar lentigo, actinic keratoses, superficial basal cell carcinoma and Kaposi's sarcoma. Cryosurgery is a safe regimen with only a few adverse effects and contraindications. Pain during and/or shortly after treatment, bulla formation and local oedema are the major, temporary adverse effects; lesional hypopigmentation and/or peripheral hyperpigmentation is the most common by occurring long-term complication. PMID- 9854155 TI - Porokeratosis and immunosuppression. PMID- 9854157 TI - Elements of the interleukin-1 signaling system show hair cycle-dependent gene expression in murine skin. AB - Stringently controlled changes in the local, cytokine-mediated signaling milieu of hair follicles (HF) have been implicated as major elements of hair cycle control and several lines of clinical and experimental evidence point towards interleukin-1 (IL-1) as an important inducer of hair loss. To address the question as to whether the steady state mRNA levels of the gene expression of the IL-1 family parallel distinct phases of the murine hair cycle, we have exploited the high degree of synchrony during depilation-induced HF cycling in mice, and have determined the mRNA levels of IL-1alpha, IL-1beta, IL-1-receptor antagonist, IL-1 receptor (R)-I and IL-1-R-II by semiquantitative RT-PCR. Our results indicate that the induced murine hair cycle is associated with profound fluctuations in the steady state mRNA levels of members of the IL-1 signaling system. Most interestingly, IL-1alpha and IL-1beta transcript levels increased dramatically with the onset of spontaneous catagen (around day 18) and peaked during telogen (day 25). These fluctuations in the IL-1alpha and IL-1beta transcript levels were paralleled by substantial expression changes of the corresponding signal transducing type I IL-1 receptor. Therefore, our findings are consistent with the concept that IL-1alpha, IL-1beta, IL-1-RI and IL-1-RII are involved in the control of catagen development. PMID- 9854158 TI - UV-A1 cytotoxicity and antioxidant defence in keratinocytes and fibroblasts. AB - The levels of antioxidant molecules and lipid peroxidation, under basal conditions, were measured in normal, human cutaneous keratinocytes and fibroblasts. Total glutathione, glutathione peroxidase and superoxide dismutases are significantly higher in normal keratinocytes compared to normal fibroblasts (respectively +248%, +193% and +155%). Under the same conditions, lipid peroxidation is significantly lower in basal keratinocytes compared to fibroblasts. UV-A1 cytotoxicity was investigated in both cutaneous cell types showing that diploid keratinocytes are more resistant to UV-A1 oxidative stress than fibroblasts (by a factor of around 8). We studied the same parameters in two keratinocyte cell lines, NCTC2544 and HaCaT cells, and in MRC5 fibroblasts. Antioxidant content and lipid peroxidation under basal conditions are quite different in these cell lines compared to those of the normal corresponding cells. Furthermore, NCTC2544 keratinocytes are more sensitive to UV-A1 radiation than normal keratinocytes whereas HaCaT keratinocytes are more resistant, and MRC5 fibroblasts are more resistant than normal cutaneous fibroblasts. These findings suggest that (i) cultured epidermal and dermal cells have different sensitivities to UV-A1 radiation that may be linked to different antioxidant capacities and (ii) cell line response to UV-A1 radiation may differ from that of normal cells. PMID- 9854159 TI - Induction of sister chromatid exchanges in fibroblasts from normal donors and from patients with xeroderma pigmentosum after combined treatment with ultraviolet radiation and modulated low frequency electric currents. AB - Dermal fibroblasts derived from normal donors as well as from patients with xeroderma pigmentosum (XP) were exposed in vitro to ultraviolet radiation of 254 nm wavelength and to weak electric 4, 000 Hz currents modulated in amplitude with 50 Hz. Treatment with this so-called interferential current (IFC) increased the rate of sister chromatid exchanges (SCE) in all fibroblast types up to roughly 50% above the spontaneous level. When IFC was applied in combination with UV radiation, it lowered UV-induced increase of the SCE-rate. A reduction of UV induced SCEs was seen when XP-cells were exposed to IFC prior to UV-treatment, compared to the reverse order of treatment. The order of exposure has only been analysed in 5 XP patients, not in normal donors. The relationships between different combinations of exposure parameters (IFC-amperage, UV-dose, cell pathology, order of treatment) were examined by analysis of variance (ANOVA). PMID- 9854160 TI - Successful treatment of disseminated facial verrucae with contact immunotherapy. AB - Disseminated flat warts are a common therapeutic problem in immunocompromised patients. However, disseminated infection on the face is, even in the immunocompetent host, a challenge. We report on a 14-year-old girl who had suffered from increasing eruptions of multiple disseminated verrucae planae et filiformes of the face for 18 months. As an alternative to silk-touch CO2-laser, contact cryotherapy or systemic immune enhancer we administered topical immunotherapy with diphenylcyclopropenone (DCP). The initial challenge with DCP 2% was performed on the forehead and the challenge was repeated weekly with DCP 0.01%. After four applications, the girl reported peeling of single warts on the forehead but more importantly at distant sites which had not received DCP. After eight weeks the patient's face was free of lesions. So far, the patient has remained free from relapse. This is the first case report of successful topical immunotherapy with DCP in disseminated facial verrucae planae and should be regarded as an effective therapeutic tool in this indication, emphasizing the great advantage of prevention of scarring of the skin induced by other therapeutic modalities. PMID- 9854161 TI - Porphyria cutanea tarda associated with human immunodeficiency virus infection. AB - Since 1987, about 60 cases of porphyria cutanea tarda (PCT) associated with human immunodeficiency virus (HIV) have been reported. The respective roles of HIV and toxic hepatic factor in PCT remain unclear. We report 10 new cases and analyse the following toxic hepatic factors: hepatitis C and B, alcoholism, drugs. The route of HIV transmission to these 10 men were: IV drugs abuse (3), homo/bisexuality (4), heterosexuality (1), and unknown (2). When PCT was diagnosed, their average age was 38 years (29-54) and the HIV-infection had been established for 4.8 years (0.33-9). Seven men had HIV-related symptoms and a CD4+ lymphocyte count below 200/mm3. Cutaneous signs and urinary porphyrin count were characteristic. Alcohol abuse was present in 8/10 patients. AST, ALT and/or gamma GT were high in 9/10 patients; 5/10 patients had HCV antibodies (4 were HCV-PCR positive). HBs antigenemia was negative among the 5/8 patients with HBV antibodies; 10/10 patients took prescribed hepatotoxic drugs. Our series confirms the presence of toxic hepatic factors in PCT of HIV-positive patients. Hepatitis C, alcoholism and hepatotoxic drug consumption seem to be triggers for the appearance of PCT in HIV-positive patients. PMID- 9854162 TI - Papular mucinosis associated with scleroderma. AB - A 49-year-old Japanese woman had been suffering from limited cutaneous scleroderma with papular mucinosis. Papular mucinosis was characterized by multiple, asymptomatic, elevated, skin-colored papules on the dorsal regions of the hands. Histopathological findings of the hard papules showed a marked deposition of hyaluronic acid along sclerosis in the middle and the lower parts of dermis. Serological studies revealed a positive antinuclear antibody (speckled type). Intravenous administration of prostaglandin E1 derivatives reduced the size of the papules, the degree of sclerodactyly and the severity of her Raynaud's phenomenon. These observations suggest that manifestations of scleroderma could be found in some cases of papular mucinosis. PMID- 9854163 TI - Pure cutaneous relapsing langerhans cell histiocytosis in an adult. AB - The histiocytic syndromes represent a group of rare diseases characterized by the proliferation of the mononuclear phagocyte system cells. We report a case of recurrent class I histiocytosis with exclusive cutaneous localization in an adult. Our patient had a 3 cm ulcerated nodule located on the right cheek. Subsequently, new lesions appeared, all having the same clinical evolution characterized by spontaneous resolution with scarring. After a five year follow up no more lesions were observed. The histopathological examination, immunohistochemical profile and electron microscopy of the lesions indicated Langerhans cell histiocytosis. PMID- 9854165 TI - Granulomatous mycosis fungoides. AB - We report a new observation of granulomatous mycosis fungoides. The diagnosis was able to be made only after performing multiple biopsies during the course of the disease. Initial evolution was rapidly favourable with electrontherapy. A granulomatous reaction is, except in Hodgkin's disease, a rare phenomenon in lymphoproliferative disorders, particularly in cutaneous T cell lymphoma. This variant of mycosis fungoides raises the problem of the histological differentiation from other granulomatous dermatoses, mainly sarcoidosis. Its prognostic significance is disputed and its pathogenesis remains unknown. PMID- 9854164 TI - Sweet's syndrome associated with granulocyte colony-stimulating factor. AB - We present two patients who developed Sweet's syndrome (SS) during recombinant granulocyte colony-stimulating factor (G-CSF) treatment. Case 1 : on day 11 of the fifth cycle of G-CSF treatment, a 21-year-old man with relapsed, intracranial germ cell tumor had fever, and painful, subcutaneous nodules on his right arm and right leg concomitant with neutrophilia. Skin biopsy revealed neutrophilic panniculitis. The skin lesions disappeared completely after discontinuance of G CSF. Case 2 : on day 7 of G-CSF treatment, a 50-year-old woman with small cell lung cancer developed fever, and widely disseminated pruritic erythema on her trunk and extremities. The histopathology of the skin was compatible with SS. Her skin lesions also disappeared after discontinuance of G-CSF treatment. She subsequently received three cycles of additional G-CSF treatments without recurrence of SS. It is possible that G-CSF treatment accidentally induced or augmented the proliferation and differentiation of clonal neutrophils with abnormal functions, since in the cases presented SS developed only once in spite of several treatments with G-CSF. PMID- 9854166 TI - Lymphangiosarcoma of the pubic region: a rare complication arising in congenital non-hereditary lymphedema. AB - Lymphangiosarcoma is a rare, aggressive, vascular neoplasm arising in chronic congenital or acquired lymphedema. Although it is most frequently associated with post-mastectomy lymphedema (Stewart-Treves's syndrome), lymphangiosarcoma can exceptionally arise in congenital hereditary lymphedema (Milroy's syndrome and Meige's syndrome) and non-hereditary lymphedema (congenital, praecox or forme tarde lymphedemas). We report a case of lymphangiosarcoma of the pubic region, supported by immunohistochemical studies, in a 42-year-old woman affected by congenital, non-hereditary lymphedema of the left genital region and homolateral lower limb. In addition, molecular analysis demonstrated the absence of Kaposi's sarcoma-associated Herpes virus (KSHV) DNA sequences in tumour lesions. To our knowledge, this is the first case of lymphangiosarcoma associated with congenital non-hereditary lymphedema confined to the pubic region. The literature concerning the cases of lymphangiosarcoma arising in congenital hereditary and non hereditary lymphedema is reviewed. Moreover, we emphasized the importance of regular clinical controls in all patients affected by chronic lymphedema. In fact, although the prognosis of this neoplasm is very poor, a prompt diagnosis and a rapid, ablative surgery associated with radiation therapy can increase the possibility of survival of these patients. PMID- 9854167 TI - Recurrent painful hand crisis in a four-year-old girl, revealing an erythropoietic protoporphyria. AB - We report the case of a 4-year-old girl, who had been suffering for 2 years from a recurrent, painful crisis affecting both hands, following sun exposure. There were no obvious cutaneous lesions, which initially caused us to consider a diagnosis of a psychiatric disorder. However, the diagnosis of erythropoietic protoporphyria was then established by the demonstration of elevated levels of free protoporphyrin in erythrocytes. The present case illustrates the effectiveness of beta-carotene associated with canthaxanthin in erythropoietic protoporphyria. PMID- 9854168 TI - A specific cutaneous lesion revealing myelodysplastic syndrome. AB - We report on an 86-year-old man with an ulcerated nodule on his left lower leg. Peripheral blood examination and bone marrow findings were compatible with the refractory anemia with an excess of blasts in transformation (RAEB-T) which is typical of the myelodysplastic syndrome (MDS). Because histological examination showed an infiltration of atypical cells of myeloid origin, this lesion was diagnosed as a specific lesion of MDS. Sometimes, only a subjective symptom, such as a skin lesion, precedes the diagnosis of MDS. PMID- 9854169 TI - Palmoplantar filiform hyperkeratosis with Darier's disease. Association or coincidence? AB - Palmoplantar orthokeratotic filiform hyperkeratosis is characterized by multiple, spiny, keratotic projections. We report a new case, occurring in an 85-year-old woman, associated with typical Darier's disease involving her chest and groin, that may be not fortuitous. Three previous reports point out its possible association with underlying diseases, such as malignancy or Darier's disease. PMID- 9854170 TI - Allergic contact dermatitis from 2-hydroxyethyl methacrylate in an adhesive on an electrosurgical earthing plate. AB - A highly (meth)acrylate-allergic patient underwent surgery because of nodular struma. Three days after her operation she developed an itching dermatitis on her left thigh. She came to our attention 18 days after the operation, because of an oozing, highly pruritic dermatitis, 8 x 19 cm in width on her left thigh, at the site where an electrosurgical earthing plate had been used during the surgery. It was revealed that the pressure-sensitive adhesive of the pad contained 2 hydroxyethyl methacrylate (2-HEMA) to which the patient earlier had had an allergic patch test reaction. The patient was negative on patch testing to other (meth)acrylates present in the pad. Patients should be questioned about possible methacrylate sensitivity before methacrylate-containing electrosurgical earthing plates are used during surgery. Allergic contact dermatitis caused by acrylic adhesives is briefly reviewed. PMID- 9854171 TI - The role of corticotropin-releasing factor in depression and anxiety disorders. AB - Corticotropin-releasing factor (CRF), a 41 amino acid-containing peptide, appears to mediate not only the endocrine but also the autonomic and behavioral responses to stress. Stress, in particular early-life stress such as childhood abuse and neglect, has been associated with a higher prevalence rate of affective and anxiety disorders in adulthood. In the present review, we describe the evidence suggesting that CRF is hypersecreted from hypothalamic as well as from extrahypothalamic neurons in depression, resulting in hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis and elevations of cerebrospinal fluid (CSF) concentrations of CRF. This increase in CRF neuronal activity is also believed to mediate certain of the behavioral symptoms of depression involving sleep and appetite disturbances, reduced libido, and psychomotor changes. The hyperactivity of CRF neuronal systems appears to be a state marker for depression because HPA axis hyperactivity normalizes following successful antidepressant treatment. Similar biochemical and behavioral findings have been observed in adult rats and monkeys that have been subjected to early-life stress. In contrast, clinical studies have not revealed any consistent changes in CSF CRF concentrations in patients with anxiety disorders; however, preclinical findings strongly implicate a role for CRF in the pathophysiology of certain anxiety disorders, probably through its effects on central noradrenergic systems. The findings reviewed here support the hypothesis that CRF receptor antagonists may represent a novel class of antidepressants and/or anxiolytics. PMID- 9854173 TI - The effect of pregnancy on the expression of uterine oxytocin, oestrogen and progesterone receptors during early pregnancy in the cow. AB - The expression of oxytocin receptor (OTR) in the uterine endometrium plays an important role in the initiation of luteolysis. During early pregnancy, the conceptus secretes interferon tau (IFN|gt) which inhibits OTR up-regulation and luteolysis. In this study, uterine horn cross sections were collected on day 16 from 15 pregnant cows (PREG), 9 uninseminated controls and 5 inseminated cows with no embryo present. The latter two groups had similar results and were combined to form a single non-pregnant (NP) group. The animals were given an oxytocin challenge shortly before tissue collection to assess prostaglandin F2alpha (PGF2alpha) release through the measurement of the metabolite 13,14 dihydro-15-keto PGF2alpha (PGFM). The mRNAs for OTR, oestrogen receptor (ER) and progesterone receptor (PR) were localised by in situ hybridisation. The results were quantified by optical density (OD) measurements from autoradiographs using image analysis. OTR protein was measured by autoradiography with iodinated oxytocin antagonist and ER and PR protein was detected by immunocytochemistry. The release of PGFM after the oxytocin challenge was significantly higher in the 14 NP cows (187%+/-15%) compared with the PREG group (131%+/-11%) (P<0.01). Low concentrations of OTR mRNA were localised to the luminal epithelium (LE) in 6 out of the 14 NP cows, of which 2 also expressed OTR protein, while OTR mRNA and protein were undetectable in all the pregnant animals. These results indicated that the sampling time coincided with the onset of the luteolytic mechanism in the NP cows. On day 16 ER mRNA was detectable in both the LE and glands of both PREG and NP animals. There were no differences in either ER mRNA or protein between NP and PREG samples. PR mRNA was moderately expressed in the caruncular stroma, with lower levels in the dense caruncular-like stroma and glands. There were no differences between PREG and NP animals. The expression of PR mRNA and protein in the deep glands was variable between animals. These results suggested that, in cows, the presence of an embryo suppressed the expression of OTR, but had no effect on the expression of the transcriptionally regulated ER on day 16. PMID- 9854172 TI - Expression and localization of the renin-angiotensin system in the rat pancreas. AB - The possibility of an intrinsic renin-angiotensin system (RAS) in the pancreas has been raised by previous studies in which immunohistochemical examination showed the presence of angiotensin II and its receptor subtypes, type 1 (AT1) and type 2 (AT2). In the present study, gene expression of several key RAS components was investigated by reverse-transcription PCR. mRNA expression for angiotensinogen, renin and angiotensin II receptor subtypes, AT1a, AT1b and AT2 was shown. The presence of angiotensinogen protein, the mandatory component for an intrinsic RAS, was demonstrated by Western blotting and localized by immunohistochemistry to the epithelia and endothelia of pancreatic ducts and blood vessels respectively. Immunoblot analysis detected a predominant protein band of about 60 kDa in the pancreas. This was consistent with the predicted value for angiotensinogen as reported in other tissues. Together with previous findings, the present study shows that the rat pancreas expresses the major RAS component genes, notably angiotensinogen and renin, required for intracellular formation of angiotensin II. The data support the notion of an intrinsic RAS in the rat pancreas which may play a role in the regulation of pancreatic functions. PMID- 9854174 TI - Indirect assessment of pulsatile gonadotropin-releasing hormone release in agonadal prepubertal rhesus monkeys (Macaca mulatta). AB - The major purpose of this study was to characterize the open-loop frequency of pulsatile GnRH release in the female rhesus monkey at an age (15-20 months) when the prepubertal restraint on the hypothalamic-pituitary axis is maximally imposed. Additionally, evidence for pulsatile GnRH release in agonadal males of comparable age was also sought. Episodic LH secretion from the pituitary was used as an indirect index of GnRH discharges. In order to maximize the sensitivity of this in situ bioassay, the responsiveness of the pituitary gonadotrophs was usually first heightened by an i.v. intermittent infusion of the synthetic peptide. Monkeys (five females, three males) were castrated between 9 and 14 months of age, implanted with indwelling venous catheters, fitted with nylon jackets and housed in specialized cages that permitted remote access to the venous circulation with minimal restraint and without interruption of the light darkness cycle. In females, LH secretion was generally assessed at 20-day intervals during alternate nighttime (1900-0200 h) and daytime (0700-1400 h) windows. In males, LH was assessed less frequently and only at night. The mean frequency of pulsatile LH release in agonadal prepubertal females was 4 pulses/7 h during the night and 2 pulses/7 h during the day. These findings indicate that, prior to puberty in the female monkey, the GnRH pulse generator operates at a relatively slow frequency and is subjected to diurnal modulation. In males, evidence for robust pulsatile GnRH release was not observed. The striking difference in activity of the GnRH pulse generator in agonadal prepubertal male and female monkeys reinforces the view that the ontogeny of the hypothalamic drive to the pituitary-gonadal axis in higher primates, including man, is sexually differentiated. PMID- 9854176 TI - Tissue-specific response of IGF-I mRNA expression to obesity-associated GH decline in the male Zucker fatty rat. AB - Diminished GH secretion is a well known association of obesity. As in obese humans, Zucker fatty rats develop a progressive GH deficiency, present at 6 weeks of age and maximal at 10 to 12 weeks. The aim of this study was to investigate the GH dependence of IGF-I gene expression in liver and extrahepatic tissues of the obese Zucker rat as a model of progressive GH reduction during adult life. Six- and 11-week-old obese Zucker rats and their lean littermates were used to compare body weight, glycemia, insulinemia, serum GH and IGF-I levels and IGF-I mRNA expression in liver, heart, aorta, kidney and skeletal muscle. In comparison with lean controls, obese Zucker rats showed at both ages comparable glycemia, severe hyperinsulinemia (mU/ml, mean+/-s.e.m.; 6 weeks 138+/-10 vs 45+/-6 P<0.001; 11 weeks 147+/-14 vs 46+/-3, P<0.001) and lower GH (ng/ml; 6 weeks 1.7+/ 0.9 vs 2.7+/-1.1; 11 weeks 1.5+/-0.9 vs 4.2+/-1.2) in the presence of similar circulating IGF-I levels (ng/ml; 6 weeks 774+/-26 vs 694+/-28; 11 weeks 1439+/ 182 vs 1516+/-121). Hepatic IGF-I mRNA expression was already reduced at 6 weeks of age due to a significant decrease in the IGF-Ib transcript compared with lean controls (relative units; IGF-Ia: 99+/-2% vs 100+/-5%; IGF-Ib: 69+/-10% vs 100+/ 2%, P<0.05) and this reduction was more marked in 11-week-old animals when both IGF-I transcripts were significantly diminished (relative units; IGF-Ia: 80+/-6% vs 100+/-1%, P<0.05; IGF-Ib: 65+/-5% vs 100+/-2%, P<0.01). Extrahepatic tissues expressed almost exclusively the IGF-Ia transcript, the amount of which relative to controls was: (1) similar at 6 weeks and decreased at 11 weeks in kidney and skeletal muscle extracts (relative units; kidney: 6 weeks 88+/-10% vs 100+/-2%; 11 weeks 76+/-3% vs 100+/-4%, P<0.05; vastus lateralis: 6 weeks 95+/-7% vs 100+/ 10%; 11 weeks 59+/-4% vs 100+/-2%, P<0.001); (2) similar at both ages in thoracic aorta (relative units; 6 weeks 121+/-6% vs 105+/-5%; 11 weeks: 91+/-14% vs 100+/ 4%); and (3) increased at both ages in left ventricle extracts (relative units; 6 weeks 114+/-2% vs 99+/-9%, P<0. 05; 11 weeks 119+/-7% vs 95+/-3%, P<0.05). specific dependence of IGF-I mRNA on GH levels during adulthood, reflected by the different behavior of IGF-I expression for each tissue in conditions of progressive decrease of GH levels. PMID- 9854175 TI - Local renin-angiotensin system contributes to hyperthyroidism-induced cardiac hypertrophy. AB - We have reported previously that thyroid hormone activates the circulating and tissue renin-angiotensin systems without involving the sympathetic nervous system, which contributes to cardiac hypertrophy in hyperthyroidism. This study examined whether the circulating or tissue renin-angiotensin system plays the principal role in hyperthyroidism-induced cardiac hypertrophy. The circulating renin-angiotensin system in Sprague-Dawley rats was fixed by chronic angiotensin II infusion (40 ng/min, 28 days) via mini-osmotic pumps. Daily i.p. injection of thyroxine (0.1 mg/kg per day, 28 days) was used to mimic hyperthyroidism. Serum free tri-iodothyronine, plasma renin activity, plasma angiotensin II, cardiac renin and cardiac angiotensin II were measured with RIAs. The cardiac expression of renin mRNA was evaluated by semiquantitative reverse transcriptase-polymerase chain reaction. Plasma renin activity and plasma angiotensin II were kept constant in the angiotensin II and angiotensin II+thyroxine groups (0.12+/-0.03 and 0.15+/-0.03 microgram/h per liter, 126+/-5 and 130+/-5 ng/l respectively) (means+/-s.e.m.). Despite stabilization of the circulating renin-angiotensin system, thyroid hormone induced cardiac hypertrophy (5.0+/-0.5 vs 3.5+/-0.1 mg/g) in conjunction with the increases in cardiac expression of renin mRNA, cardiac renin and cardiac angiotensin II (74+/-2 vs 48+/-2%, 6.5+/-0.8 vs 3.8+/-0.4 ng/h per g, 231+/-30 vs 149+/-2 pg/g respectively). These results indicate that the local renin-angiotensin system plays the primary role in the development of hyperthyroidism-induced cardiac hypertrophy. PMID- 9854177 TI - Tri-iodothyronine inhibits multilayer formation of the osteoblastic cell line, MC3T3-E1, by promoting apoptosis. AB - Cell death through apoptosis is a well-known mechanism for maintaining homoeostasis in many developmental and pathological processes. We have recently presented evidence for the occurrence of apoptosis during the formation of bone like tissue in vitro. MC3T3-E1 osteoblast-like cells in culture develop features of the osteoblastic phenotype and form many cell layers embedded in extracellular matrix which can mineralise. Tri-iodothyronine (T3), even though it enhances the expression of many osteoblastic features, attenuates the multilayer formation to about two layers. The aim of this study was to investigate how T3 prevents multilayer formation. MC3T3-E1 cells were seeded at different densities and cultured for up to 2 weeks. Thereafter we analysed proliferation rate and the distribution of the phases of the cell cycle and studied apoptosis. We found that T3 did not inhibit DNA synthesis. Analysis of the cell cycle phases showed an increase in the number of cells in G0/G1 with increasing cell density, but no significant effect of T3 treatment was found. Morphological investigations showed apoptotic features in both cell layers and culture supernatants. The cells exhibited typical plasma membrane blebbings, chromatin condensation, DNA fragmentation and phagocytosed apoptotic bodies. T3 treatment significantly increased the number of apoptotic cells. We conclude from our data that T3 inhibits multilayer formation of MC3T3-E1 cells by increasing the rate of apoptosis and not by inhibition of proliferation. Because apoptosis is a fundamental regulatory event during bone tissue differentiation, our findings emphasise the importance of thyroid hormones in bone maintenance and development. PMID- 9854178 TI - Interleukin-1alpha regulates G1 cell cycle progression and arrest in thyroid carcinoma cell lines NIM1 and NPA. AB - This study provides the first report that the same cytokine (interleukin-1 (IL 1)) can induce opposite effects on cyclin-dependent kinases (Cdks) and Cdk inhibitors (Cdkis) in the G1 phase even in the same type of cancer cells (papillary thyroid carcinoma cells). Cell cycle analysis revealed an increase in NIM1 cells and a decrease in NPA cells in the S and G2+M phases after treatment with IL-1alpha. The addition of IL-1alpha to NIM1 cells reduced the expression of p16 and p21 protein and induced the expression of Cdk2 and Cdk4 protein, which leads to the phosphorylation of retinoblastoma protein. The addition of IL-1alpha to NPA cells induced the expression of p27 protein and reduced the expression of Cdk2 protein, which leads to induction of p107 protein expression. It is of interest that p21 protein expression was not observed in NPA cells. These results suggest that several Cdks and Cdkis play a regulatory role in the G1 cell cycle progression and arrest induced by IL-1alpha in thyroid carcinoma cell lines. PMID- 9854179 TI - Hormonal changes during parturition in heifers and goats are related to the phases and severity of labour. AB - Parturition is a natural event that involves stress and pain for the mother. We thus hypothesized that levels of stress hormones measured during parturition could reflect levels reached in response to severe discomfort and pain of other kinds as well. The aim of this study was therefore to determine whether plasma concentrations of cortisol, adrenaline, noradrenaline, beta-endorphin, met enkephalin, vasopressin and oxytocin vary depending on the phase and severity of labour in dairy heifers (ten) and dairy goats (six), and how these hormones interact with each other. Blood samples were taken once a day for 3 days before labour and for 3 days afterwards and at predetermined phases during labour. All heifers delivered one calf and five of them needed obstetrical assistance. Two of the goats delivered one kid, and four had twins; all kidded without help. The cortisol concentration peaked when the calf and the first kid were born. In the heifers, plasma adrenaline increased after delivery, while the noradrenaline concentration did not change significantly in heifers that needed assistance, but increased during expulsion in heifers calving without help. In the goats, adrenaline and noradrenaline concentrations increased in association with expulsion of the first kid. The beta-endorphin concentration increased during labour in goats. In heifers that needed assistance, beta-endorphin concentration increased 1 h after labour but there was no change in heifers that did not need assistance. The met-enkephalin concentration was elevated during expulsion in heifers and fluctuated in the goats. Both oxytocin and vasopressin increased during expulsion in both groups of heifers, but vasopressin increased four times more in heifers needing assistance. In the goats, oxytocin reached its highest levels just as the feet of the first kid became visible, and vasopressin peaked as the head emerged. Parturition took longer in heifers that needed assistance than in those that did not. It is concluded that, even though the pattern of change differed between hormones during labour, the changes were related to the phases of labour. A longer labour therefore meant that the hormone concentrations stayed elevated for longer. Vasopressin reached high levels in goats and was the only hormone for which plasma concentrations were higher in heifers that needed assistance than in those that did not, indicating that this hormone is released in order to deal with the pain-related stress associated with labour. PMID- 9854180 TI - Influence of vitamin D3 deficiency and 1,25 dihydroxyvitamin D3 on de novo insulin biosynthesis in the islets of the rat endocrine pancreas. AB - Because 1,25 dihydroxyvitamin D3 (1,25(OH)2D3) is known to activate the biosynthesis of numerous proteins in various tissues, experiments were undertaken to compare the influence of 1,25(OH)2D3 in vitro on both the secretion and biosynthesis of insulin in islets of Langerhans from both 4-week vitamin D3 deficient rats and normal rats. Islets were either incubated or perifused after a 6-h induction period in the presence of various concentrations of 1, 25(OH)2D3 from 10(-12) M, which was inactive in controls, to 10(-6) M. Experiments were performed in the presence of a non-labelled amino acid mixture, to favour protein synthesis. Tritiated tyrosine was added as tracer during glucose stimulation. The newly synthesised proteins, labelled with [3H]tyrosine, were extracted by an acid alcohol method and separated by gel chromatography adapted for low-molecular weight proteins. Even in the presence of the amino acid mixture, the insulin response of the islets to 16.7 mM glucose was decreased by vitamin D3 deficiency and improved by 1,25(OH)2D3. This beneficial effect did not occur in basal conditions, but only during glucose stimulation, and was observed in both phases of insulin release. Moreover, these effects disappeared in the presence of 5x10( 4 )M cycloheximide, a protein biosynthesis inhibitor. Islets from vitamin D3 deficient rats exhibited a general decrease in the amount of de novo biosynthesised proteins and of [3H]tyrosine-labelled insulin and proinsulin fractions. A 6-h period of 1,25(OH)2D3 induction significantly improved the amount of de novo biosynthesised proteins, and particularly of newly synthesised insulin in response to a 2-h glucose stimulation. Calculation of the rate of conversion of newly synthesised proinsulin-like material to insulin as the [3H]insulin/[3H]proinsulin-like material ratio provided evidence for a dose dependent increase, induced by 1, 25(OH)2D3, that could exceed that of normal islets. These data support the hypothesis that 1,25(OH)2D3 in vitro not only facilitated the biosynthetic capacity of the beta cell - which was highly induced during a 16.7-mM glucose stimulation, via a global activation of islets protein biosynthesis - but also produced an acceleration of the conversion of proinsulin to insulin. PMID- 9854181 TI - Thyroid hormone stimulates basal and interleukin (IL)-1-induced IL-6 production in human bone marrow stromal cells: a possible mediator of thyroid hormone induced bone loss. AB - It is well known that excessive thyroid hormone in the body is associated with bone loss. However, the mechanism by which thyroid hormone affects bone turnover remains unclear. It has been shown that it stimulates osteoclastic bone resorption indirectly via unknown mediators secreted by osteoblasts. To determine if interleukin-6 (IL-6) or interleukin-11 (IL-11) could be the mediator(s) of thyroid hormone-induced bone loss, we studied the effects of 3,5,3'-tri iodothyronine (T3) on basal and interleukin-1 (IL-1)-stimulated IL-6/IL-11 production in primary cultured human bone marrow stromal cells. T3 at 10(-12)-10( 8) M concentration significantly increased basal IL-6 production in a dose dependent manner. It also had an additive effect on IL-1-stimulated IL-6 production, but failed to elicit a detectable effect on basal or IL-1-stimulated IL-11 production. Treatment with 17beta-estradiol (10(-8) M) did not affect the action of T3 on IL-6/IL-11 production. These results suggest that thyroid hormone may stimulate bone resorption by increasing basal and IL-1-induced IL-6 production from osteoblast-lineage cells, and these effects are independent of estrogen status. PMID- 9854182 TI - Interactions between oestradiol and glucocorticoid regulatory effects on liver specific glucocorticoid-inducible genes: possible evidence for a role of hepatic 11beta-hydroxysteroid dehydrogenase type 1. AB - In vitro, 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD-1) catalyses the interconversion of active corticosterone and inert 11-dehydrocorticosterone. 11beta-HSD-1 is highly expressed in liver, where the reaction direction is 11beta reduction, thus potentially increasing intrahepatic active glucocorticoid levels. Inhibition of 11beta-HSD-1 increases insulin sensitivity in humans in vivo suggesting that hepatic 11beta-HSD-1 plays a role in the maintenance or control of key glucocorticoid-regulated metabolic functions. We have selectively repressed hepatic 11beta-HSD-1 in rats by oestradiol administration for 42 days. This nearly completely repressed hepatic 11beta-HSD-1 mRNA expression and enzyme activity and reduced expression of hepatic glucocorticoid-inducible genes including phosphoenolpyruvate carboxykinase (PEPCK), the rate-limiting step in gluconeogenesis. Similar effects were seen after 3 weeks of oestradiol treatment. To examine whether this was due to any direct effect of oestradiol upon PEPCK, the experiment was repeated in adrenalectomised rats+/-glucocorticoid replacement. In adrenalectomised rats, oestradiol did not attenuate hepatic PEPCK, whilst glucocorticoid replacement restored this action. Oestradiol did not alter hepatic metabolism of corticosterone by pathways other than 11beta-HSD-1. These data suggest 11beta-HSD-1 plays an important role in maintaining expression of key glucocorticoid-regulated hepatic transcripts. Enzyme inhibition may provide a useful therapeutic target for manipulating glucose homeostasis. PMID- 9854183 TI - Expression of 17beta-hydroxysteroid dehydrogenase types 1, 2, 3 and 4 in the human temporal lobe. AB - Sex steroid hormones exert important biological effects on the brain. Moreover, an extensive sex steroid metabolism occurs in the brain. In sex steroid metabolism 17beta-hydroxysteroid dehydrogenases (17beta-HSDs) play essential roles in catalyzing the final steps in androgen and estrogen biosynthesis. Recently four types of human 17beta-HSDs and a pseudogene of the type 1 isoform were identified. To date, 17beta-HSD has not been extensively studied in the human brain. Therefore, we investigated the mRNA expression of the four isozymes of 17beta-HSD as well as the pseudogene of the type 1 isoform in the human temporal lobe to determine the predominant isoforms and, moreover, to elucidate the existence of possible sex and age differences. We studied biopsy materials from the temporal lobe of 34 women, 32 men and 10 children. Quantification of different mRNAs was achieved by competitive reverse transcription-PCR. 17beta-HSD 1, 17beta-HSD 3 and 17beta-HSD 4 were expressed in the human temporal lobe of children and adults, whereas 17beta-HSD 2 and the pseudogene of 17beta-HSD 1 were not expressed. In adults, 17beta-HSD 3 and 17beta-HSD 4 mRNA concentrations were significantly higher in the subcortical white matter (17beta-HSD 3: 14 591+/-3457 arbitrary units (aU), mean+/-s.e.m.; 17beta-HSD 4: 1201+/-212 aU) than in the cortex (17beta-HSD 3: 5428+/-1057 aU, P<0. 0002; 17beta-HSD 4: 675+/-74 aU, P<0.004). 17beta-HSD 1 concentrations did not differ significantly between the white matter (3860+/-1628 aU) and the cortex (2525+/-398 aU) of adults. In conclusion, the present study demonstrates the expression of 17beta-HSD 1, 3 and 4 mRNAs in the human temporal lobe. Together with CYP19AROM and 5alpha-reductase, known to be expressed in the human brain, the expression of 17beta-HSD 1, 3 and 4 mRNAs indicates the major importance of local steroid biosynthesis in the brain. PMID- 9854184 TI - Insulin action in growth hormone-deficient and age-matched control rats: effect of growth hormone treatment. AB - The isolated effect of growth hormone on carbohydrate metabolism in rat skeletal muscle was studied in growth hormone-deficient dwarf rats (dw/dw) treated with either recombinant human growth hormone or saline for 10 days. In addition, age matched heterozygous (DW/dw) (normal weight and plasma IGF-I) control rats were treated with saline. Growth hormone increased weight gain from 0.1+/-0.1 (s.e.m) to 3.6+/-0.1 g/day and plasma IGF-I concentration from 364+/-23 to 451+/-32 ng/ml. Glucose metabolism in skeletal muscle perfused with basal, submaximal and maximal concentrations (0, 600 and 60 000 pmol/l respectively) of insulin was not changed by growth hormone. No change could be detected in the total number of glucose transporters (GLUT1 and GLUT4) in the skeletal muscles, except from a lower amount of GLUT4 in the soleus muscle in the heterozygous control group. However, at submaximal insulin concentrations, skeletal muscle glucose uptake and transport were significantly lower in the heterozygous control group compared with the growth hormone-deficient group. This could indicate either a direct long term effect of growth hormone or more likely a secondary effect attributable to the difference in body weight (205.2+/-3.1 vs 361. 6+/-5.9 g for dwarf rats and heterozygous controls respectively), and thereby muscle fibre size, between the groups probably resulting in lower average interstitial insulin and glucose concentrations at a given plasma concentration in the heterozygous rats. It is concluded that restoration of subnormal growth hormone concentrations for 10 days has no effect on insulin-stimulated glucose metabolism in skeletal muscle in vitro. PMID- 9854185 TI - Involvement of the rapamycin-sensitive pathway in the insulin regulation of muscle protein synthesis in streptozotocin-diabetic rats. AB - Insulin resistance in 3-day streptozotocin (STZ)-treated rats was manifested by the lack of antiproteolytic action of insulin as well as by a reduction of its stimulatory effect on protein synthesis (-60% compared with the control group) in epitrochlearis muscle incubated in vitro. In the present study, we have investigated the diabetes-associated alterations in the insulin signalling cascade, especially the phosphatidylinositol-3 kinase (PI-3 kinase)/p70 S6 kinase (p70(S6K)) pathway, in rat skeletal muscle. LY 294002, a specific inhibitor of PI 3 kinase, markedly decreased the basal rate of protein synthesis and completely prevented insulin-mediated stimulation of this process both in control and diabetic rats. Thus, PI-3 kinase is required for insulin-stimulated muscle protein synthesis in diabetic rats as in the controls. Rapamycin, an inhibitor of mammalian target of rapamycin (mTOR), had no effect on the basal rate of protein synthesis in either of the experimental groups. In control rats, the stimulatory action of insulin on muscle protein synthesis was diminished by 36% in the presence of rapamycin, whereas in diabetic muscles this reduction amounted to 68%. The rapamycin-sensitive pathway makes a relatively greater contribution to the stimulatory effect of insulin on muscle protein synthesis in diabetic rats compared with the controls, due presumably to the preferential decrease in the rapamycin-insensitive component of protein synthesis. Neither basal nor insulin stimulated p70(S6K) activity, a signalling element lying downstream of mTOR, were modified by STZ-diabetes. PMID- 9854186 TI - Cyclic AMP regulates expression of the gene coding for a mouse vas deferens protein related to the aldo-keto reductase superfamily in human and murine adrenocortical cells. AB - Mouse vas deferens protein (MVDP) is a member of the aldo-keto reductase superfamily. The regulation of MVDP gene expression by activators of the protein kinase A signalling pathway was investigated in human (H295-R) and murine (Y1) adrenocortical carcinoma cells. Immunoblotting with polyclonal antibodies showed that MVDP is expressed in adrenal glands from mouse, rat, rabbit and guinea-pig, probably under the control of ACTH. In both adrenocortical cell lines used, MVDP is constitutively synthesized and its accumulation is increased by treatment with cAMP or forskolin. MVDP mRNA steady-state levels were up-regulated by forskolin in adrenocortical cells by a process that does not require de novo protein synthesis. The results suggest that cAMP is at least one of the key regulators of adrenal MVDP expression and that this effect is direct. PMID- 9854187 TI - Seasonal changes in the negative feedback regulation of the secretion of the gonadotrophins by testosterone and inhibin in rams. AB - Three experiments were conducted with castrated Romney Marsh rams (wethers) to investigate the ability of testosterone and inhibin to suppress the secretion of LH and FSH during the breeding and the non-breeding seasons. In Experiment 1, wethers (n=5/group) were treated every 12 h for 7 days with oil or 16 mg testosterone propionate (i.m.) and were then given two i.v. injections either of vehicle or of 0.64 microg/kg human recombinant inhibin A (hr-inhibin) 6 h apart. Blood samples were collected for 4 h before inhibin or vehicle treatment and for 6 h afterwards for the assay of LH and FSH. In Experiments 2 and 3 wethers underwent hypothalamo-pituitary disconnection (HPD) and were given 125 ng GnRH i.v. every 2 h. In Experiment 2, HPD wethers (n=3/group) were injected (i.m.) every 12 h with oil or testosterone and blood samples were collected over 9 h before treatment and 7 days after treatment. In Experiment 3, HPD (n=5/group) wethers were treated with vehicle or hr-inhibin, as in Experiment 1, after treatment with oil, or 4, 8 or 16 mg testosterone twice daily for 7 days. Blood samples were collected over 4 h before treatment with vehicle or hr-inhibin and for 6 h afterwards. Treatment of wethers with testosterone (Experiment 1) resulted in a significant decrease in the plasma concentrations of LH and number of LH pulses per hour but the magnitude of these reductions did not differ between seasons. Testosterone treatment had no effect on LH secretion in GnRH pulsed HPD wethers in either season and treatment with hr-inhibin did not affect LH secretion in wethers or HPD wethers in any instance. Plasma concentrations of FSH were significantly (P<0.05) reduced following treatment with testosterone alone during the breeding season but not during the non-breeding season. FSH levels were reduced to a greater extent by treatment with hr-inhibin but this effect was not influenced by season. During the non-breeding season, the effect of hr-inhibin to suppress FSH secretion was enhanced in the presence of testosterone. These experiments demonstrate that the negative feedback actions of testosterone on the secretion of LH in this breed of rams occurs at the hypothalamic level and is not influenced by season. In contrast, both testosterone and inhibin act on the pituitary gland to suppress the secretion of FSH and these responses are affected by season. Testosterone and inhibin synergize at the pituitary to regulate FSH secretion during the non-breeding season but not during the breeding season. PMID- 9854188 TI - Molecular diagnostics and therapy of prostate cancer: new avenues. AB - Co-operation and communication between clinicians and scientists is required to meet the major challenges presented by the diagnosis and therapy of prostate cancer. Molecular oncology is playing an increasing role in this field and has already been instrumental in elucidating many of the basic mechanisms underlying the development and progression of prostate cancer. By understanding these mechanisms, factors which determine whether the tumour will metastasise, such as loss of function of E-cadherin, have been identified and may help the clinician determine which therapeutic strategy is most appropriate for an individual patient. Clinicians also need more sensitive tools to help them diagnose prostate cancer and monitor its progression. The marker DD3/PCA3 shows potential in this respect. Perhaps the most fruitful area for molecular research is in the definition of new therapeutic targets useful in hormone-refractory prostate cancer. In the early stages of development are those agents which target the activation of programmed cell death, inhibition of signal transduction, inactivation of telomerase activity, and differentiation therapy. In order to accelerate the implementation of diagnostic aids and more effective therapeutic strategies for prostate cancer, clinicians must have a greater insight into the molecular mechanisms operating in their patients' disease and scientists need to understand the clinical problems involved. PMID- 9854189 TI - Orchidectomy and oestrogen therapy revisited. AB - Over the past 20 years therapeutic options for prostate cancer have increased. Nevertheless, there may still be a role for long-established treatments such as orchidectomy and oestrogens. Orchidectomy is a simple surgical procedure, and patient survival is comparable with other treatments involving androgen ablation. However, loss of libido and sexual function is an expected outcome and hot flushes occur in about 50% of patients. Osteoporosis, loss of muscle mass, and the psychological impact associated with orchidectomy are of concern, particularly with increasing treatment periods. Nevertheless, orchidectomy is indicated when an immediate reduction of testosterone levels is required, or the patient does not comply with other treatments or objects to the cost of medical therapy. Oestrogen therapy may be superior to castration in terms of efficacy, but orally administered oestrogens are associated with gynaecomastia, loss of sexual function and unacceptable cardiovascular toxicity. Low dose oestrogens in combination with antiandrogens or antithrombotic agents may be better tolerated treatments. The route of administration is a crucial factor in the genesis of cardiovascular toxicity and parenterally administered oestrogens may not entail the same risk. Further research in this area is warranted. PMID- 9854190 TI - Antiandrogens as monotherapy for prostate cancer. AB - Castration or antiandrogen monotherapies remain options for prostate cancer treatment as only marginal benefits have been demonstrated with combined androgen blockade, although it may be that certain subgroups of patient may benefit. Of the nonsteroidal antiandrogens, bicalutamide 150 mg was as effective as castration in M0 patients with significant improvement in sexual interest and physical capacity, but the trial has yet to reach maturity. In M1 patients, bicalutamide 150 mg was not as effective as castration but this may be outweighed by symptomatic and quality of life benefits. Nilutamide is not recommended as monotherapy and there are little data on flutamide. The steroidal antiandrogen, cyproterone acetate, is as effective as oestrogen therapy and has a better side effect profile, although cardiovascular and hepatic side effects are still of concern. Compared with flutamide, in a recently completed EORTC study, side effects such as gynaecomastia, diarrhoea, nausea, and liver function deterioration occurred less often, and thrombotic effects more often, in the cyproterone acetate group. No difference was seen in the preservation of sexual functioning. Quality of life issues are becoming increasingly important and thus antiandrogen monotherapy may become more widely used in the management of prostate cancer. PMID- 9854191 TI - Endocrine withdrawal syndrome and its relevance to the management of hormone refractory prostate cancer. AB - Therapeutic options for patients with metastatic prostate cancer relapsing from primary hormonal therapy are limited. On the selective discontinuation of flutamide in patients that have relapsed on combined androgen ablation, a third of the patients will show a significant clinical benefit for 4-6 months. A multivariate model has identified prolonged exposure to combined androgen blockade, high baseline alkaline phosphatase and prolonged flutamide exposure as prognostic factors for patients that have a significant prostate-specific antigen (PSA) decline after the withdrawal of flutamide. This phenomenon has also been described with bicalutamide and other antiandrogens, and thus has been more appropriately renamed the endocrine withdrawal syndrome. The molecular basis for this endocrine withdrawal syndrome is not completely understood but data suggest that mutations in the androgen receptor may be responsible for the paradoxical effect observed. Recognition of this syndrome has introduced a non-toxic therapy for advanced prostate cancer patients and has had a dramatic impact on the interpretation and design of clinical trials in patients with 'hormone refractory disease'. PMID- 9854192 TI - Arguments for the long-term use of combined androgen blockade. AB - From what is known about the testicular and adrenal origins of testosterone, and the testosterone dependency of prostate cancer, treatment with combined androgen blockade (CAB) by castration plus an antiandrogen was expected to be beneficial to patients. A number of early studies verified this hypothesis, but the findings needed to be confirmed in large, randomized, prospective studies. Modest but significant benefits of CAB were seen when treatment with leuprorelin plus flutamide was compared with leuprorelin alone. Of note was the increased benefit from CAB seen in patients with minimal metastatic prostate cancer, a finding that needed to be confirmed. A study comparing orchidectomy plus nilutamide with orchidectomy alone showed significant benefits of CAB in terms of time to progression and overall survival. However, a recent major NCI study revealed no benefits for CAB with respect to survival, even for patients with minimal metastatic disease. Results from other clinical studies have been conflicting, perhaps because of the method of castration or differences in antiandrogen used. Before the question of whether CAB significantly benefits patients can be fully answered, a clearer understanding of the interactions between antiandrogens, luteinizing hormone-releasing hormone agonists and receptors, and of the response of prostate cancer to these, is needed. PMID- 9854193 TI - Arguments against the long-term use of combined androgen blockade. AB - Androgen deprivation therapy is the most effective systemic treatment for advanced prostate cancer. However, as most patients who die from prostate cancer have hormone refractory disease, fine tuning of antiandrogen treatment by combined androgen blockade (CAB) can not be expected to improve survival significantly. Only the South West Oncology Intergroup (SWOG) study 0036 has shown a significant advantage for CAB compared with luteinizing hormone-releasing hormone (LH-RH) agonist alone. However, the results of this study should be interpreted with caution as the patients had to self-administer their treatment by daily injection so compliance may not have been optimal. Also, those receiving LH-RH agonist alone were not covered against disease flare. Indeed, no trial using depot LH-RH agonist with or without flutamide has been able to show a survival benefit. When treatment with LH-RH agonist plus antiandrogen was compared with orchiectomy alone, only the European Organization for Research and Treatment of Cancer (EORTC) study 30,853 showed a significant difference in favour of CAB. However, in this study an increased proportion of patients receiving CAB may have had a more favourable prognosis. Only one study comparing orchiectomy plus antiandrogen with orchiectomy alone has shown an advantage for CAB therapy, and this was only slight. Therefore, as yet there is no justification for long-term use of CAB. However, short-term antiandrogen treatment must be used to prevent disease flare during initiation of LH-RH agonist treatment. PMID- 9854194 TI - Therapeutic dilemmas in prostate cancer: justification for watchful waiting. AB - Despite the large numbers of elderly men known to harbour pathological evidence of prostate cancer and evidence from observational studies suggesting the prolonged time course of such disease, conservative management of such patients has not enjoyed the widespread support of urologists, particularly in the USA, where monitoring of patients is often dismissed as 'intentional neglect' rather than 'watchful waiting'. This paper outlines the rationale and principles underlying expectant treatment of patients with prostate cancer. PMID- 9854195 TI - Intermittent androgen suppression for prostate cancer: rationale and clinical experience. AB - The rationale behind intermittent androgen suppression (IAS) is based on: (1) observations that androgen ablation is palliative, not curative, in most patients with prostate cancer, and that quality of life must be considered; (2) the assumption that immediate androgen ablation is superior to delayed therapy in improving survival; (3) the hypothesis that if tumour cells surviving androgen withdrawal are forced into a normal pathway of differentiation by androgen replacement, then apoptotic potential might be restored and progression to androgen independence delayed. Several centres have now tested the feasibility of IAS therapy in non-randomized groups of prostate cancer patients using serum of prostate-specific antigen levels as trigger points. Clinical data suggest that prostate cancer is amenable to control by IAS and offers clinicians an opportunity to improve patients' quality of life by balancing the benefits of immediate androgen ablation (delayed progression and prolonged survival) while reducing treatment-related side effects and expense. Whether time to progression and survival is affected in a beneficial or adverse way is being studied in randomized, prospective protocols. PMID- 9854196 TI - [Diagnosis and therapy of special movement disorders]. PMID- 9854197 TI - [Management of schizophrenic psychoses: a one-year experience with Serdolect]. PMID- 9854198 TI - [Is the systematic exploration of the common bile duct justified during laparoscopic cholecystectomy?]. PMID- 9854199 TI - [Laparoscopic cholecystectomy and lithiasis of the common bile duct: prospective study on the importance of preoperative endoscopic ultrasonography and endoscopic retrograde cholangiography]. AB - OBJECTIVES: Laparoscopic cholecystectomy is the standard treatment of symptomatic gallstones. At present, no consensus has been reached on the diagnostic and therapeutic methods of concomitant common bile duct stones. Systematic preoperative endoscopic ultrasonography followed, if necessary, by endoscopic retrograde cholangiography and sphincterotomy during the same anesthetic procedure could be a diagnostic and therapeutic alternative for common bile duct stones making possible a laparoscopic cholecystectomy without intraoperative investigation of the common bile duct. METHODS: One hundred and twenty-five patients underwent a prospective endoscopic ultrasonographic evaluation prior to laparoscopic cholecystectomy for symptomatic gallstones. Fourty-four patients (35%) had at least one predictive factor for common bile duct stones. Endoscopic ultrasonography and cholecystectomy were performed on the same day. Endoscopic ultrasonography was followed by endoscopic retrograde cholangiography and sphincterotomy by the same endoscopist in case of common bile duct stones on endoscopic ultrasonography. Patients were routinely followed up between 3 and 6 months and one year after cholecystectomy. RESULTS: Endoscopic ultrasonography suggested common bile duct stones in 21 patients (17%). Endoscopic ultrasonography identified a stone in 17 of 44 patients (38.6%) with predictor of common bile duct stones and only in 4 of 81 patients (4.9%) without predictor of common bile duct stone. Among these 21 patients, one patient was not investigated with endoscopic retrograde cholangiography because of the high risk of sphincterotomy, 19 patients had a stone removed after sphincterotomy, one patient had no visible stone neither on endoscopic retrograde cholangiography, nor on exploration of the common bile duct after sphincterotomy. Endoscopic ultrasonography was normal in 104 patients (83%). However, two patients in this group were investigated with endoscopic retrograde cholangiography because endoscopic ultrasonography was incomplete in one case and because endoscopic ultrasonography was normal in the second case but a stone in the left hepatic duct was detected by ultrasonography. A stone was removed after endoscopic sphincterotomy in these two patients. In the group of 102 patients without stone, 91 out of 92, continued to be asymptomatic during a median follow-up of 8.5 months. One patient with symptoms one month after cholecystectomy underwent endoscopic sphincterotomy but no stone was found. CONCLUSIONS: Systematic preoperative endoscopic ultrasonography followed, if necessary with endoscopic retrograde cholangiography and sphincterotomy is a diagnostic and therapeutic alternative for common bile duct stones making possible a laparoscopic cholecystectomy without intraoperative investigation of the common bile duct for all patients. This alternative is only justifiable in patients with predictor of common bile duct stones. PMID- 9854200 TI - [Creation of a viral hepatitis B and C in the Cote-d'Or department. Methodology and initial results]. AB - OBJECTIVES: The epidemiological status of viral B and C hepatitis remains unknown in the general French population. This is why a specialized registry was created in the French Cote-d'Or administrative area (493,931 inhabitants) on 1994 January 1st. METHODS: The three sources of information were: a) biological and virological analysis laboratories, which report all new HBs Ag and HCV Ab cases, b) specialists in gastroenterology, hepatology, infectious diseases and internal medicine, c) pathologists. Additional information was obtained from the medical practitioner who prescribed the serology. RESULTS: Between January and December 1994, 241 new cases of HBs Ag and HCV Ab were reported: 168 cases of HCV Ab (96 males, 72 females) and 73 cases of HBs Ag (37 males, 36 females) including 10 mixed cases (HBs Ag and HCV Ab). The annual standardized rate of detection of patients with HCV Ab was 26.4 +/- 4.6/100,000 inhabitants. Contamination was a result of intravenous drug addiction in 54 cases (32%), blood transfusion in 39 cases (23%) and remained unknown in 56 cases (33%). A liver biopsy was performed in 42 patients, 17 were treated with interferon during the two years following diagnosis. The standardized detection rate of patients with HBs Ag was 12.9 +/- 2.6/100,000 inhabitants. Contamination resulted from sexual transmission in 17 cases (23%), was related to a lengthy stay in endemic countries in 10 cases (14%) or to intravenous drug addiction in 6 cases (8%), and remained unknown in 37 cases (51%). A liver biopsy was performed in 6 cases and 2 patients were treated. CONCLUSION: These preliminary French population based data show that the annual frequency of detection of HBs Ag and HCV Ab is high and that care of these patients must be improved. PMID- 9854201 TI - [Assigning the cause of diarrhea and enterocolitis to drugs. Current approach and outlook for improvement]. PMID- 9854202 TI - [Causal assessment of drug-induced acute colitis. A prospective study of 58 consecutive cases]. AB - AIMS: The causal effect of drugs is underestimated in patients with inflammatory bowel disease. The aim of this study was to assess the causal implication of drugs in acute colitis. METHODS: A prospective study was conducted in 58 consecutive patients with an acute inflammation of the colonic mucosa. Recent drug intake was recorded and possible causal effects were analyzed exhaustively with respect to both intrinsic and bibliographic criteria. RESULTS: Causal assessment scores were high for 57 drugs and 41 patients. Drug-induced acute colitis was diagnosed in 35 cases. In 7 patients, physician practice had not taken into account drug use despite probable drug involvement. The main drugs implicated were antibiotics (n = 42) and non steroidal anti-inflammatory drugs (n = 10). CONCLUSION: Acute colitis is mainly induced by drugs. PMID- 9854203 TI - [Descriptive study of digestive functional symptoms in the French general population]. AB - To study the prevalence of "reported" functional digestive symptoms (FDS) in terms easily understood by the general population without resorting to predefined concepts of functional syndromes, and to assess FDS impact on public health, a sample survey has been carried out between September and December 1995. METHODS: Four thousand eight hundred and seventeen subjects representative of the French general population aged 15 years or more filled in a questionnaire describing their digestive disorders. RESULTS: Seventy percent of the subjects had digestive complaints, 9% being related to a presumably organic disease, and 61% attributed to FDS. Twenty-seven % of the subjects claimed to be inconvenienced by their FDS, whereas 34% seemed not to feel any inconvenience. Among FDS, gas emission was the most frequent symptom (59%), followed by stomach ache and/or digestive pain (48%), flatulence (47%), bad digestion sensations (40%), constipation (35%), aerophagia (29%), bad breath (22%), incomplete evacuation of stools (19%). FDS had lasted from 6 months to 5 years in 38%, and over 5 years in 52%. In the subgroup of subjects inconvenienced by FDS (27%), 9% consulted and 18% did not, whereas in the subgroup not inconvenienced, 3% consulted and 31% did not. Altogether, 26% of the subjects followed a prescription or self medication treatment; 35% were not treated. Some explanatory variables appeared to be associated with the onset of inconvenience: the associations pain and bad digestion, flatulence and aerophagia, incomplete evacuation and nervous or presumably organic origin of FDS, age, stress, FDS frequency. Duration of symptoms, age above 65 years, digestive pain, presumably organic origin, and FDS frequency were associated with the need to consult. This descriptive, pragmatic survey shows the widespread prevalence of FDS, affecting 28 million French people. Functional digestive disorders in the "academic" meaning constitute only a limited subset. FDS lead to major health care consumption. Their impact on public health is undoubtedly greater than the estimates derived from studies designed in accordance with conventional nosological categories. PMID- 9854204 TI - [Effect of serotonin on unidirectional ion fluxes in rat intestine in vivo]. AB - AIMS: A stimulating intestinal secretory effect is described in vitro and an inhibition with selective inhibitors of the different receptors of serotonin (5 HT), in vivo. But a direct effect, in vivo, in fully vascularized and innervated intestine has not yet been clearly evidenced. We studied the effect of 5-HT in anesthetized rats with ligated loops. This work, performed at 4 intestinal levels, allowed a comparison with the effects of a known stimulant of intestinal secretion, VIP, and a specific inhibitor of Na/H exchange, dimethylamiloride (DMA). RESULTS: 5-HT induced an inhibition of epithelial Na influx in agreement with the inhibition of Na/H exchanger, an inhibition of the influx of Cl, partially passive absorption following Na by paracellular route. A decrease of Na and Cl efflux was induced by 5-HT in duodenum, jejunum and ileum while in colon, a stimulation was obtained by intraluminal but not intravenous route. CONCLUSION: Even though 5-HT induced a liquid accumulation in all intestinal segments, the effect differed according to the intestinal level, either inhibition of absorption in the small intestine, or stimulation of secretion in the colon. The comparison of the effect of 5-HT with that of DMA shows that the inhibition of absorption is not only due to Na/H exchanger inhibition. PMID- 9854205 TI - [Effectiveness and safety of lansoprazole in the treatment of Zollinger-Ellison syndrome. First six months of treatment]. AB - OBJECTIVES: The aim of this prospective study was to confirm the efficacy and safety of lansoprazole in patients with Zollinger-Ellison syndrome (ZES). METHODS: Fourteen patients (5 W, 9 M) with ZES, age (mean +/- SD) 55.5 +/- 12.8 years, were included in the study. STUDY DESIGN: initially and at 1, 3 and 6 months thereafter the following items were assessed: clinical signs, fasting serum gastrin (FSG), basal acid output (BAO) before next dose of lansoprazole. BAO < 10 mmol H+/h was considered as efficient. Initially and at 6 months, laboratory tests (hematology, liver, renal and hormonal), endoscopy and histological enterochromaffin-like cell and gastrin cell density assessments were performed. Lansoprazole initial dose was adjusted according to clinical symptoms and secretory studies. RESULTS: At 6 months, lansoprazole doses of 60, 90, 120 and 180 mg/d maintained BAO < 10 mmol H+/h in 9, 2, 1 and 1 patient, respectively. No significant changes in FSG, endocrine cells densities and biological parameters were noted during treatment. Neither adverse events nor carcinoid tumors were observed. We conclude that lansoprazole is efficient and well tolerated in patients with ZES. PMID- 9854206 TI - [Imaging of somatostatin receptors in gastroenteropancreatic neuroendocrine tumors]. PMID- 9854207 TI - [Primary neuroendocrine tumor in the intrahepatic bile ducts: a rare finding in an asymptomatic patient]. AB - Neuroendocrine tumors of the biliary tree are rare. In all cases except one, diagnosis was made in symptomatic patients. We report a case of an asymptomatic intrahepatic bile duct neuroendocrine tumor in a 74-year old man. To our knowledge, this is the second reported case of an asymptomatic intrahepatic bile duct neuroendocrine tumor. Diagnosis was only made by anatomopathological examination of the tumor after resection. Systemic and immunohistochemical hormonal screening was negative. Twenty months after surgery, the patient was asymptomatic and there was no recurrence. PMID- 9854208 TI - [Cystic and mucinous lesion in an antral ectopic pancreas]. AB - We report a case of heterotopic pancreas located in the gastric antrum. The cystic formation contained mucus. Tumoral markers in the cyst fluid were within the range pancreatic cystic mucinous tumors. Pathology examination of the resected specimen did not evidence any proliferative lesions but showed papillary hyperplasia probably due to duct occlusion. PMID- 9854209 TI - [Familial varices of the colon. Report of four cases]. AB - We report four cases of ileo-colonic varices in one family. This congenital disease is uncommon with only 21 cases described in the literature. Recurrent digestive tract bleeding is the most common manifestation, but varices can be asymptomatic. Digestive endoscopy leads to the diagnosis. The diagnosis of familial colonic varices can be considered when colonic varices occur in several members of a family with no evident cause, particularly portal hypertension or venous thrombosis. No treatment is usually necessary except when major or recurrent bleeding indicates surgical resection. These malformations are classified as vascular ectasia in the subgroup of cavernous haemangiomas. PMID- 9854210 TI - [Dexchlorpheniramine-induced acute hepatitis: a case with positive rechallenge]. PMID- 9854211 TI - [Severe iron-deficiency anemia due to diffuse antral vascular ectasia in a cirrhosis patient. Cure after surgical portacaval shunt]. PMID- 9854212 TI - [Rifamycin-induced cholangitis with positive rechallenge test]. PMID- 9854213 TI - [Hypoglycemia secondary to IGF-II hypersecretion in the course of metastatic colonic adenocarcinoma]. PMID- 9854214 TI - [Bronchogenic cyst of the cervical esophagus]. PMID- 9854215 TI - [First familial form of Crohn's disease in Reunion Island]. PMID- 9854216 TI - [Importance of tomodensitometry for the detection of a foreign body of vegetable nature responsible for a jejunal occlusion]. PMID- 9854217 TI - [Ischemic colitis after taking flutamide]. PMID- 9854218 TI - [Molecular analysis of the progression of Helicobacter pylori-associated chronic gastritis toward mucosa-associated lymphoid tissue (MALT) lymphoma]. PMID- 9854219 TI - [The information system on perinatality in France. Situation at the national level]. PMID- 9854220 TI - [Monitoring of the course of labor: how to monitor childbirth 25 years after the institution of monitoring]. PMID- 9854221 TI - [Is vaginal hysterectomy important for large uterus of more than 500 g? Comparison with laparotomy]. AB - OBJECTIVE: Vaginal hysterectomy is an advantageous surgical technique as compared with abdominal hysterectomy: operating time is shorter, it is safer and hospitalization stay and recovery time are shorter. However in France, 2/3 of all hysterectomies are still performed by laparotomy. Would the vaginal approach be reasonable if a difficult hysterectomy is expected? The purpose of our study was to compare the per- and postoperative complications and the period following vaginal hysterectomy versus the abdominal route in patients with a large uterus. PATIENTS AND METHODS: Forty-nine vaginal hysterectomies were compared retrospectively with 22 abdominal hysterectomies for fibroma during the period from 01.01.91 to 31.12.95. Uterine weight in all cases was between 500 to 1,000 g. RESULTS: The average uterine weight was significantly different between the vaginal and the abdominal groups (644 g vs 747 g, p = 0.02). Operating time, pre- and postoperative complications and analgesic use were not significantly different between the two groups. Operative bleeding was significantly higher in the abdominal group than in the vaginal group (659 ml vs. 359 ml, p = 0.006), as well as hemoglobin loss (2.49 vs. 1.82, p = 0.04). There was also a statistical difference between the vaginal group and the abdominal groups in terms of hospital stay (4 d vs. 6 d respectively, p = 0.0002). CONCLUSION: Vaginal hysterectomy with morcellation is advantageous in comparison with abdominal hysterectomy even when the uterine weight is over 500 g, since it offers better post-operative comfort than laparotomy without endangering the patients. The main contraindications for the vaginal route are uterus beyond the ombilicus and vaginal atresia. Excepting these cases, the indication for the vaginal approach depends on the patient's past surgical history, uterine mobility and vaginal access. In more difficult cases, these factors are assessed under anesthesia. PMID- 9854222 TI - [Hysteroscopic myomectomy. Functional results with an average follow-up of 6 years]. AB - OBJECTIVE: In 1995, we presented our experience in hysteroscopic resection of hemorrhagic submucous fibroids in 196 patients. The objective of the present work was to analyze functional outcome three years later. PATIENTS: One hundred ninety six patients with abnormal uterine bleeding were treated between 1987 and 1993. Equipment, techniques, characteristics of the procedures and early results were detailed in the previous article. RESULTS: With a mean follow-up of 73 months (range 50,104), results were: 13.8% of the patients were lost to follow-up; 68.4% had symptomatic improvement and failure was observed in 17.8% (subsequent hysterectomy in 12.7%). Forty-nine patients had repeat resection in this series; 61 became menopausal after surgery, and 21 were taking hormone replacement therapy with good results. CONCLUSION: Developed as an alternative to hysterectomy, operative hysteroscopy has proven to be a safe and effective procedure. This treatment modality appears to give satisfactory long-term results, with a low rate of complications. PMID- 9854223 TI - [Evaluation of the association of a serum marker and second trimester ultrasonography for the screening of trisomy 21 in women of less than 38 years. Prospective study of 5,163 patients]. AB - OBJECTIVES: Use of serum markers alone for trisomy 21 screening programs leads to a high rate of amniocentesis. Adding a second parameter (ultrasonography during the second trimester) might reduce this rate yet retain satisfactory sensitivity. This work was conducted to evaluate the pertinence of associating serum hCG level between 16 and 17 weeks gestation and morphological ultrasonography between 18 ans 20 weeks gestation. METHOD: A prospective study was conducted in 5,163 pregnant women aged over 38 years. A morphological ultrasonography was performed in all patients whose hCG level > 1/150 indicated a risk. An amniocentesis was proposed if an anomaly was detected at ultrasonography. RESULTS: Serum hCG was above the risk threshold retained in 11.9% of the patients and among these patients at least one anomaly was detected at ultrasonography in 12%. An amniocentesis was performed in 1.4% of all patients. The positive predictive value of the screening test was 20%. CONCLUSION: Combining a serum marker and ultrasonography during the second trimester allows a reduction in the rate of amniocentesis compared with screening with serum markers alone. Sensitivity for detecting trisomy 21 remains satisfactory. PMID- 9854225 TI - [Screening of post-partum depressive states in the Yalgado Ouedraogo National Hospital Center maternity ward in Ouagadougou Burkina Faso]. AB - OBJECTIVES: Depression is a frequent early complication of the post-partum period. It affects the mother's own psychological function and has an important impact on the mother-child relation. PATIENTS AND METHODS: We recruited a successive series of 34 women (mean age 27.4 +/- 4.5 years) who delivered at the Yalgado Ouedraogo University Hospital maternity ward. Patients were evaluated with the Beck Depression Inventory on day 3 and 8 post-partum and a questionnaire was used to record social, demographic and sanitary data. RESULTS: Average Beck Depression Inventory scores on day 3 and 8 were 9.3 +/- 4.5 and 13.5 +/- 6.2. In our series, the state of depression was the same on day 3 and 8 in 78.6% of the mothers. Mood disorders were significantly correlated with parity and the sex of the child. Other variables studies (age, occupation, matrimonial status, mode of nursing) did not influence significantly the development of a mood disorder. CONCLUSION: The psychosocial environment plays a determining role in the development of depressive states. Adequate management requires cooperative care by the obstetrician, psychiatrist and pediatrician. PMID- 9854224 TI - [Critical analysis of hemostasis disorders in the course of eclampsia. Report of 106 cases]. AB - OBJECTIVE: Study of hemostatic disorders during eclampsia, their risk factors, maternal complications and associated mortality. METHODS: Retrospective study concerning 106 cases of severe eclampsia treated in intensive care between September 1992 and December 96. Patients with or without hemostatic disorders were compared for laboratory findings, maternal complications and mortality. RESULTS: Forty patients had hemostasis disorders as follows: isolated thrombopenia in 19 cases, disseminated intravascular coagulation (DIC) in 5 cases, Hellp syndrome associated to DIC in 7 cases and Hellp syndrome in 9 cases. Hemostasis disorders were associated to maternal advanced age, but not with gestational age or blood pressure in admission or time of convulsions. Complications and mortality associated with hemostasis disorders were more frequent compared to patients without hemostasis disorders. Among the 17 deaths of our series, 10 had hemostasis disorders. CONCLUSION: Hemostasis disorders were prognosis factors in eclampsia requiring systematic laboratory tests at admission and immediate delivery. PMID- 9854226 TI - [Ursodeoxycholic acid: prospect for treatment of gravidic cholestasis? Report of 3 cases]. AB - Ursodeoxycholic acid, employed in treatment of intrahepatic cholestasis as seen in primary biliary cirrhosis, primary sclerosing cholangitis, and chronic hepatitis; does not have marketing approval for prescription during pregnancy because of lack of data. In 3 cases of gravidic cholestasis, we administered oral ursodeoxycholic acid 1 g a day from the 34th week of amenorrhea to delivery. In each case, it took 3 days of treatment for the pruritus to regress incompletely and for plasma levels of biliary acid and transaminases to decrease. The infants, born between the 36th and 38th week of amenorrhea, presented with no problem. Forty-eight cases of gravidic cholestasis treated by ursodeoxocholic acid (0.4 to 1 g a day) have been reported in the literature; 18 cases belonging to 2 randomized studies. In 46 cases pruritus disappeared generally 3 days after treatment onset, and plasma level of biliary acid and transaminase decreased in one week. Only two patients experienced persisting pruritus despite biological improvement. No foetal adverse effect is reported. Ursodeoxycholic acid seems to be an efficient treatment of gravidic cholestasis. Long term observation of fetuses exposed in utero to this treatment is required to assess safety. PMID- 9854227 TI - [Pheochromocytoma associated with pregnancy. Report of 2 cases and review of the literature]. AB - Pheochromocytoma is rarely observed during pregnancy and not easily diagnosed, especially since the clinical manifestations may mimic common gravid hypertension. The maternal and fetal prognosis depends on early diagnosis and multidisciplinary management preparing for tumor resection. Appropriate laboratory tests easily provide the positive diagnosis, if they are ordered. The tumor is localized by ultrasonography or magnetic resonance imaging (MRI). By prescribing an alpha-blocker, it is possible to prepare the resection as a function of term, before or after delivery. This strategy allows a reduction in maternal and fetal mortality which is high if the pheochromocytoma goes undiagnosed. PMID- 9854228 TI - [Congenital heart disease and nuchal translucency with normal karyotype. Report of 3 cases]. AB - We report three pregnancies where enlarged nuchal translucency was discovered at the first trimester transvaginal ultrasound examination; congenital heart disease developed later. Two cases of hypoplastic left heart were diagnosed prenatally at the mid-trimester sonographic examination. The pregnancies were terminated. In the third case, a supravalvular pulmonary stenosis was discovered on the second day of life. Further investigations demonstrated a mutation on the elastin locus, thus confirming the diagnosis of Williams-Beuren syndrome. The role of nuchal translucency as a risk marker for congenital heart disease is discussed. PMID- 9854229 TI - [Parital cutaneous necrosis of the abdominal wall after cesarean section]. AB - The authors report an original case of large cutaneous necrosis of the abdominal wall following a first cesarean section in a patient with a background of pemphigoid gestationis. There was no direct skin burn nor infection of the dermis and this necrosis is typical of venous suffering. The mechanism is debated, probably the thrombosis of the superficial inferior epigastric vein due to the spreading out of a monopolar coagulation current. A one-step treatment was done using a wide excision and an abdominal advancement flap. This surgical technique allowed a fast cicatrisation and led to a very good long-term esthetical result. PMID- 9854230 TI - [Readmission in a Maternity Hospital for early mother-child relationship disorders]. AB - We report the original experience of admission in a post-delivery care unit, of a mother and her one-month-old child, for the treatment of post-delivery depression, in association with mother-child relational disorders. In this emergency situation, a maternity hospital team was involved in a maternity care. PMID- 9854231 TI - Financing children's health care needs in the next century. PMID- 9854232 TI - Opportunities to improve health care for low-income adolescents. PMID- 9854234 TI - Health care coverage and access for children in an urban state: the New York perspective. PMID- 9854233 TI - Medicaid's impact on access to and utilization of health care services among racial and ethnic minority children. PMID- 9854235 TI - Child development and pediatrics for the 21st century: the healthy steps approach. PMID- 9854236 TI - Access denied: taking action for medically underserved children. PMID- 9854237 TI - Children with chronic conditions in the 21st century. AB - The care of children who have chronic conditions should focus on improving the lives of the children and their families. This is accomplished most effectively through community-based, family-centered care. Critical issues for these children and families in entering the next century include the level of insurance, content of benefits package, level of evidence required for health insurance reimbursement, preparation of and incentives for health care professionals, roles being demanded of families. As the richest country in the world, we should be able to (1) provide universal coverage, (2) guarantee a comprehensive benefits package that applies a child-specific standard of medical necessity, (3) establish criteria for requiring insurers to reimburse for new effective therapies, (4) develop strategies for overcoming professional barriers, and (5) create mechanisms for compensating family caretakers for their care. PMID- 9854238 TI - Have professional recommendations and consumer demand altered pediatric practice regarding child development? AB - OBJECTIVE: Amid growing consumer demand and professional society recommendations for more information on early childhood development, current practices of pediatricians in regard to children's development remain largely unknown. We investigate whether there are differences in provider practices and satisfaction with regard to children's development (based on length of time in practice). DESIGN: A self-reported survey was conducted of physicians at 30 pediatric practices participating in the Healthy Steps for Young Children Program. Healthy Steps is a national program to enhance the developmental potential of young children. Comparisons were made among physicians categorized as in training (n = 88), recently in practice (completing residency from 1984 to 1996, n = 69), or more experienced (completing residency prior to 1984, n = 52). PRINCIPAL FINDINGS: Relative to those recently in practice and in training, more experienced pediatricians spend less time in well-baby visits in the first 2 months of life. One-third of physicians conduct family risk assessments, half complete routine developmental screening, and over half do safety risk assessments in the first 2 months of life. There were few differences by provider experience in the topics covered under anticipatory guidance for new parents. Nearly all discussed infant car seats, sleep position, feeding practices, and temperament, but less than half routinely discussed domestic violence, and between half and three-quarters discussed infant bathing, maternal depression, and appropriate discipline practices. While all three groups of physicians were satisfied with the amount of time to discuss growth and development and parenting issues, more experienced physicians were more satisfied with their own and their staff's abilities to meet new parents' needs on these issues. Factors that over one-third of physicians reported affected their ability to deliver the best quality care were shortage of support staff, limited referral sources, managed care restrictions on referrals for special services, excessive paperwork, and lack of time for follow up, teaching parents, and answering questions. Physicians in recent practice were more likely than more experienced physicians to cite reimbursement concerns and limited staff to address the needs of parents regarding development. CONCLUSIONS: Most pediatricians do not conduct routine developmental screening in the first 2 months of life, and most discuss safety, as opposed to developmental and mental health, concerns with parents of newborns. Pediatricians with more experience believe they are better meeting new parents' needs and are less likely to cite systems and organizational factors as limiting their ability to deliver high-quality care. PMID- 9854239 TI - Creating systems of developmental health care for children. PMID- 9854240 TI - Information needs in public health and health policy: results of recent studies. PMID- 9854241 TI - The retrieval problem for health policy and public health: knowledge bases and search engines. PMID- 9854242 TI - Information resources for public health practice. PMID- 9854243 TI - Web-based resources for retrieving health policy information: NLM and beyond. AB - This paper identifies Web-based resources of interest to the health policy community. Both information useful for developing health policy and information about existing health policy are included. Resources described are classified into three major categories: traditional and grey literature, statistical and epidemiological data, and legal and legislative material. National Library of Medicine (NLM) resources include MEDLINE, DIRLINE, HealthSTAR, HSRProj, and HSTAT. In addition, NLM's National Information Center on Health Services Research and Health Care Technology (NICHSR) has a Web page (http:/(/)www.nlm.nih.gov/nichsr/nichsr.++ +html) that provides an extensive listing of health-policy-related Web sites. Some of the other resources highlighted include those available from the Health Care Financing Administration (HCFA), the National Center for Health Statistics (NCHS) of the Centers for Disease Control and Prevention (CDC), the National Committee for Vital and Health Statistics (NCVHS), and the Library of Congress. PMID- 9854244 TI - Sources used in health policy research and implications for information retrieval systems. PMID- 9854245 TI - Quality of public health information: lessons from the field. PMID- 9854246 TI - Applied information quality: a framework for thinking about the quality of specific information. AB - Information quality, considered abstractly, may seem to be a relatively straightforward matter. Information should be accurate, up to date, useful, and attributable to reputable sources. However, determining the quality of a specific piece of information for a specific use is a more complicated process. The concept of applied information quality is defined in this paper as a judgment of information quality (1) made by a specific person or persons, (2) in a specific situational context for use of that information, and (3) based on the characteristics of the information. Each of the three elements of the judgment influences its outcome. Information judgments are made by individuals in the context of their discipline and community of practice. The situational context includes the specifics of the context for use of the information, the questions that the information must address, the strategy for locating potentially relevant information, and the body of information that is retrieved and is available for judgment and use. The paper focuses on the third element of a judgment of information quality--the characteristics of the information on which the judgment is based. These characteristics are grouped for discussion under six metaquestions: What is the information item of interest? How was the focal information created and when? Who is involved with the focal information? From what perspective was the information created and why? What relationships does the focal information have to other information--its antecedents, sources, and other related information? What approval, review, or other filtering processes, if any, has the information gone through? Approaches to improving quality judgments can focus on improving the information itself, improving the channels that organize and deliver information, or improving the individual's ability to judge the quality of information for a specific purpose. These are not mutually exclusive and, probably, all should be pursued. Applied judgments of information quality are ultimately the responsibility of the individuals using information; they need to be supported in this professional activity as they are in the other responsibilities of their professional practice. PMID- 9854247 TI - Meeting information needs in health policy and public health: priorities for the National Library of Medicine and The National Network of Libraries of Medicine. AB - Those seeking information in health policy and public health are not as well served as those seeking clinical information. Problems inhibiting access to health policy and public health information include the heterogeneity of professionals seeking the information, the distribution of relevant information across disciplines and information sources, scarcity of synthesized information useful to practitioners, lack of awareness of available services or training in their use, and lack of access to information technology or to knowledgeable librarians and information specialists. Since 1990, the National Library of Medicine and the National Network of Libraries of Medicine have been working to enhance information services in health policy and public health through expanding the coverage of the NLM collection, building new databases, and engaging in targeted outreach and training initiatives directed toward segments of the health policy and public health communities. Progress has been made, but more remains to be done. Recommendations arising from the meeting, Accessing Useful Information: Challenges in Health Policy and Public Health, will help NLM and the National Network of Libraries of Medicine to establish priorities and action plans for the next several years. PMID- 9854248 TI - Meeting information needs in health policy and public health: roles for the Centers for Disease Control and Prevention. PMID- 9854249 TI - Strategies for addressing priority information problems in health policy and public health. PMID- 9854250 TI - The pharmacoeconomic impact of antimicrobial therapy for peptic ulcer disease in a large urban jail. AB - PURPOSE: To determine the pharmacoeconomic impact of antimicrobial treatment of peptic ulcer disease (PUD) in a large urban jail. PATIENTS AND METHODS: Retrospective comparison of PUD-related pharmacy and laboratory expenditures over a 2-year period before and after the institution of a PUD treatment protocol with the priority of Helicobacter pylori eradication for inmates in Rikers Island Correctional Facility. RESULTS: After the protocol was adopted, total pharmacy related and laboratory-related expenses for PUD care decreased by 40.2%, and expenditures for ranitidine declined by 52.2%. There was an increase in spending for antimicrobial agents and H. pylori antibody testing, but this was insignificant compared to the savings generated by decreased ranitidine usage. Annual savings in our facility as a result of this intervention were $123,449. CONCLUSIONS: Modern therapeutic strategies for PUD aimed at eradicating H. pylori can result in significant savings in the institutional setting; these savings are largely attributable to the decreased usage of histamine-2 receptor antagonists. PMID- 9854251 TI - Supracervical and total abdominal hysterectomy trends in New York State: 1990 1996. AB - To describe practice trends for total abdominal hysterectomy (TAH) and supracervical abdominal hysterectomy (SCH) in New York State and to identify fiscal features associated with these two operations, all inpatient discharges for TAH and SCH performed for benign indications from 1990 to 1996 were reviewed using the Statewide Planning and Resource Cooperative System, a centralized data reporting system. For each year examined, the number of TAHs and SCHs performed, the procedure rates adjusted for the total New York State female population, and the per diem charge (calculated from mean institutional charge as a function of average length of stay) were evaluated. While the TAH rate declined in New York State, from 34.0 in 1990 to 28.4 in 1996 (P = .01), the SCH rate increased nearly five-fold during the same period, from 0.62 to 3.07 (P = .0003). Patients tended to be discharged later following SCH than for TAH, although by 1996, the LOS for both operations was equivalent. The per diem institutional charge for SCH was consistently higher than for TAH in each year studied. The changes in charge and relative frequency of TAH and SCH in New York State invite further study to describe these trends more fully. PMID- 9854252 TI - The use of the artificial kidney in the treatment of uremia. 1952. PMID- 9854254 TI - The role of prefrontal cortex in working memory: examining the contents of consciousness. AB - Working memory enables us to hold in our 'mind's eye' the contents of our conscious awareness, even in the absence of sensory input, by maintaining an active representation of information for a brief period of time. In this review we consider the functional organization of the prefrontal cortex and its role in this cognitive process. First, we present evidence from brain-imaging studies that prefrontal cortex shows sustained activity during the delay period of visual working memory tasks, indicating that this cortex maintains on-line representations of stimuli after they are removed from view. We then present evidence for domain specificity within frontal cortex based on the type of information, with object working memory mediated by more ventral frontal regions and spatial working memory mediated by more dorsal frontal regions. We also propose that a second dimension for domain specificity within prefrontal cortex might exist for object working memory on the basis of the type of representation, with analytic representations maintained preferentially in the left hemisphere and image-based representations maintained preferentially in the right hemisphere. Furthermore, we discuss the possibility that there are prefrontal areas brought into play during the monitoring and manipulation of information in working memory in addition to those engaged during the maintenance of this information. Finally, we consider the relationship of prefrontal areas important for working memory, both to posterior visual processing areas and to prefrontal areas associated with long-term memory. PMID- 9854253 TI - Single units and conscious vision. AB - Figures that can be seen in more than one way are invaluable tools for the study of the neural basis of visual awareness, because such stimuli permit the dissociation of the neural responses that underlie what we perceive at any given time from those forming the sensory representation of a visual pattern. To study the former type of responses, monkeys were subjected to binocular rivalry, and the response of neurons in a number of different visual areas was studied while the animals reported their alternating percepts by pulling levers. Perception related modulations of neural activity were found to occur to different extents in different cortical visual areas. The cells that were affected by suppression were almost exclusively binocular, and their proportion was found to increase in the higher processing stages of the visual system. The strongest correlations between neural activity and perception were observed in the visual areas of the temporal lobe. A strikingly large number of neurons in the early visual areas remained active during the perceptual suppression of the stimulus, a finding suggesting that conscious visual perception might be mediated by only a subset of the cells exhibiting stimulus selective responses. These physiological findings, together with a number of recent psychophysical studies, offer a new explanation of the phenomenon of binocular rivalry. Indeed, rivalry has long been considered to be closely linked with binocular fusion and stereopsis, and the sequences of dominance and suppression have been viewed as the result of competition between the two monocular channels. The physiological data presented here are incompatible with this interpretation. Rather than reflecting interocular competition, the rivalry is most probably between the two different central neural representations generated by the dichoptically presented stimuli. The mechanisms of rivalry are probably the same as, or very similar to, those underlying multistable perception in general, and further physiological studies might reveal much about the neural mechanisms of our perceptual organization. PMID- 9854255 TI - Consciousness and the structure of neuronal representations. AB - The hypothesis is defended that brains expressing phenomenal awareness are capable of generating metarepresentations of their cognitive processes, these metarepresentations resulting from an iteration of self-similar cortical operations. Search for the neuronal substrate of awareness therefore converges with the search for the nature of neuronal representations. It is proposed that evolved brains use two complementary representational strategies. One consists of the generation of neurons responding selectively to a particular constellation of features and is based on selective recombination of inputs in hierarchically structured feedforward architectures. The other relies on the dynamic association of feature-specific cells into functionally coherent cell assemblies that, as a whole, represent the constellation of features defining a particular perceptual object. Arguments are presented that favour the notion that the metarepresentations supporting awareness are established in accordance with the second strategy. Experimental data are reviewed that are compatible with the hypothesis that evolved brains use assembly codes for the representation of contents and that these assemblies become organized through transient synchronization of the discharges of associated neurons. It is argued that central states favouring the formation of assembly-based representations are similar to those favouring awareness. PMID- 9854256 TI - The neuronal basis for consciousness. AB - Attempting to understand how the brain, as a whole, might be organized seems, for the first time, to be a serious topic of inquiry. One aspect of its neuronal organization that seems particularly central to global function is the rich thalamocortical interconnectivity, and most particularly the reciprocal nature of the thalamocortical neuronal loop function. Moreover, the interaction between the specific and non-specific thalamic loops suggests that rather than a gate into the brain, the thalamus represents a hub from which any site in the cortex can communicate with any other such site or sites. The goal of this paper is to explore the basic assumption that large-scale, temporal coincidence of specific and non-specific thalamic activity generates the functional states that characterize human cognition. PMID- 9854257 TI - Consciousness and body image: lessons from phantom limbs, Capgras syndrome and pain asymbolia. AB - Words such as 'consciousness' and 'self' actually encompass a number of distinct phenomena that are loosely lumped together. The study of neurological syndromes allows us to explore the neural mechanisms that might underlie different aspects of self, such as body image and emotional responses to sensory stimuli, and perhaps even laughter and humour. Mapping the 'functional logic' of the many different attributes of human nature on to specific neural circuits in the brain offers the best hope of understanding how the activity of neurons gives rise to conscious experience. We consider three neurological syndromes (phantom limbs, Capgras delusion and pain asymbolia) to illustrate this idea. PMID- 9854259 TI - Investigating the biology of consciousness. AB - The fact that consciousness is a private, first-person phenomenon makes it more difficult to study than other cognitive phenomena that, although being equally private, also have characteristic behavioural signatures. Nonetheless, by combining cognitive and neurobiological methods, it is possible to approach consciousness, to describe its cognitive nature, its behavioural correlates, its possible evolutionary origin and functional role; last but not least, it is possible to investigate its neuroanatomical and neurophysiological underpinnings. In this brief essay I distinguish between two kinds of consciousness: core consciousness and extended consciousness. Core consciousness corresponds to the transient process that is incessantly generated relative to any object with which an organism interacts, and during which a transient core self and transient sense of knowing are automatically generated. Core consciousness requires neither language nor working memory, and needs only a brief short-term memory. Extended consciousness is a more complex process. It depends on the gradual build-up of an autobiographical self, a set of conceptual memories pertaining to both past and anticipated experiences of an individual, and it requires conventional memory. Extended consciousness is enhanced by language. PMID- 9854258 TI - Memory, consciousness and neuroimaging. AB - Neuroimaging techniques that allow the assessment of memory performance in healthy human volunteers while simultaneously obtaining measurements of brain activity in vivo may offer new information on the neural correlates of particular forms of memory retrieval and their association with consciousness and intention. We consider evidence from studies with positron emission tomography and functional magnetic resonance imaging indicating that priming, a form of implicit retrieval, is associated with decreased activity in various cortical regions. We also consider evidence concerning the question of whether two components of explicit retrieval--intentional or effortful search and successful conscious recollection--are preferentially associated with increased activity in prefrontal and medial temporal regions, respectively. Last, we consider recent efforts to probe the relation between the phenomenological character of remembering and neural activity. In this instance we broaden our scope to include studies employing event-related potentials and consider evidence concerning the neural correlates of qualitatively different forms of memory, including memory that is specifically associated with a sense of self, and the recollection of particular temporal or perceptual features that might contribute to a rich and vivid experience of the past. PMID- 9854260 TI - Towards a functional neuroanatomy of conscious perception and its modulation by volition: implications of human auditory neuroimaging studies. AB - Conscious sensory perception and its modulation by volition are integral to human mental life. Functional neuroimaging techniques provide a direct means of identifying and characterizing in vivo the systems-level patterns of brain activity associated with such mental functions. In a series of positron emission tomography activation experiments, we and our colleagues have examined a range of normal and abnormal auditory states that, when contrasted, provide dissociations relevant to the question of the neural substrates of sensory awareness. These dissociations include sensory awareness in the presence and absence of external sensory stimuli, the transition from sensory unawareness to awareness (or vice versa) in the presence of sensory stimuli, and sensory awareness with and without volition. The auditory states studied include hallucinations, mental imagery, cortical deafness modulated by attention, and hearing modulated by sedation. The results of these studies highlight the distributed nature of the functional neuroanatomy that is sufficient, if not necessary, for sensory awareness. The probable roles of unimodal association (as compared with primary) cortices, heteromodal cortices, limbic/paralimbic regions and subcortical structures (such as the thalamus) are discussed. In addition, interactions between pre- and post rolandic regions are examined in the context of top-down, volitional modulation of sensory awareness. PMID- 9854261 TI - The neural correlates of consciousness: an analysis of cognitive skill learning. AB - This paper presents a functional brain-imaging strategy designed to isolate neural correlates of consciousness in humans. This strategy is based on skill learning. In the example presented (rapidly generating verbs for visually presented nouns), a cognitive skill is examined before and after practice. As shown, there are marked qualitative differences in the neural circuitry supporting performance of this task in the naive and practised state that include, importantly, both increases and decreases from the baseline activity of the brain. PMID- 9854262 TI - Possible mechanisms of anosognosia: a defect in self-awareness. AB - Anosognosia of hemiplegia is of interest for both pragmatic and theoretical reasons. We discuss several neuropsychological theories that have been proposed to explain this deficit. Although for psychological reasons people might deny deficits, the denial hypothesis cannot account for the hemispheric asymmetries associated with this disorder and cannot explain why some patients might deny one deficit and recognize another equally disabling deficit. There is some evidence that faulty feedback from sensory deficits, spatial neglect and asomatognosia might be responsible for anosognosia in some patients. However, these feedback hypotheses cannot account for anosognosia in all patients. Although the hemispheric disconnection hypothesis is appealing, disconnection is probably only a rare cause of this disorder. The feedforward intentional theory of anosognosia suggests that the discovery of weakness is dependent on attempted action and some patients might have anosognosia because they do not attempt to move. We present evidence that supports this theory. The presence of one mechanism of anosognosia, however, does not preclude the possibility that other mechanisms might also be working to produce this disorder. Although a large population study needs to be performed, we suspect that anosognosia might be caused by several of the mechanisms that we have discussed. On the basis of the studies of impaired corporeal self-awareness that we have reviewed, we can infer that normal self awareness is dependent on several parallel processes. One must have sensory feedback and the ability to attend to both one's body and the space where parts of the body may be positioned or acting. One must develop a representation of the body, and this representation must be continuously modified by expectations (feedforward) and knowledge of results (feedback). PMID- 9854263 TI - The autonomy of the visual systems and the modularity of conscious vision. AB - Anatomical and physiological evidence shows that the primate visual brain consists of many distributed processing systems, acting in parallel. Psychophysical studies show that the activity in each of the parallel systems reaches its perceptual end-point at a different time, thus leading to a perceptual asynchrony in vision. This, together with clinical and human imaging evidence, suggests strongly that the processing systems are also perceptual systems and that the different processing-perceptual systems can act more or less autonomously. Moreover, activity in each can have a conscious correlate without necessarily involving activity in other visual systems. This leads us to conclude not only that visual consciousness is itself modular, reflecting the basic modular organization of the visual brain, but that the binding of cellular activity in the processing-perceptual systems is more properly thought of as a binding of the consciousnesses generated by each of them. It is this binding that gives us our integrated image of the visual world. PMID- 9854264 TI - Attention, self-regulation and consciousness. AB - Consciousness has many aspects. These include awareness of the world, feelings of control over one's behaviour and mental state (volition), and the notion of continuing self. Focal (executive) attention is used to control details of our awareness and is thus closely related to volition. Experiments suggest an integrated network of neural areas involved in executive attention. This network is associated with our voluntary ability to select among competing items, to correct error and to regulate our emotions. Recent neuroimaging studies suggest that these various functions involve separate areas of the anterior cingulate. We have adopted a strategy of using marker tasks, shown to activate the brain area by imaging studies, as a means of tracing the development of attentional networks. Executive attention appears to develop first to regulate distress during the first year of life. During later childhood the ability to regulate conflict among competing stimuli builds upon the earlier cingulate anatomy to provide a means of cognitive control. During childhood the activation of cingulate structures relates both to the child's success on laboratory tasks involving conflict and to parental reports of self-regulation and emotional control. These studies indicate a start in understanding the anatomy, circuitry and development of executive attention networks that serve to regulate both cognition and emotion. PMID- 9854265 TI - Coordinated expression in chronically unconscious persons. AB - The clinically described 'persistent vegetative state' (PVS), consists of wakefulness unaccompanied by any evidence of the subject's awareness of self or environment. Past studies from our own and other laboratories have used positron emission tomography (PET) to study brain metabolism in approximately 20 such patients during wakeful periods. All those efforts identified global cerebral glucose metabolism at or below levels encountered during deep barbiturate anaesthesia. Nevertheless, the clinical literature includes rare reports of relatively isolated cognitive functions expressed by PVS patients late in their course. The observation raises the question of whether such activity reflects awareness or unconscious automatic behaviour. We employed magnetometry (MEG), PET scanning, MR imaging and 24-hour EEG recordings to evaluate three patients clinically vegetative between six months and 20 years after onset. Neither meticulous clinical examinations nor 24-hour EEG and video monitoring provided any hint of cognitive interaction in any subject. Nevertheless, patient 1 uttered single words once every 48 hours or more; patient 2 frequently expressed coordinated, non-purposeful, non-dystonic movements in arms and/or legs; and, patient 3 expressed strong emotional negativity without motor responses to noxious stimuli with occasional quieting in response to prosodic stimuli. All patients had whole-brain averaged global metabolism levels below 50% of normal. Patient 1, however, demonstrated preserved islands of increased metabolism in the posterior frontal and posterior temporal lobes, as well as MEG activations of Heschl's gyrus all located in the left hemisphere. In patient 2, selected increased metabolism was confined to the frontal poles and related subcortical structures. MRI in patient 3 demonstrated severe, bilateral post-traumatic cerebral atrophy. PET metabolism was diffusely reduced to 40% of normal but MEG evoked potentials indicated early and late sensory processing with abnormal later evoked components. The correlation of fragmentary behaviour with preserved metabolic and physiologic activity in cortical and subcortical regions known to support specific modular functions is novel. The finding demonstrates the capacity of severely damaged brains to partially express surviving modular functions without evidence of integrative processes that would be necessary to produce consciousness. We conclude that the mere expression of isolated neuropsychologic activity by isolated modules is insufficient to generate consciousness in overwhelmingly damaged brains. PMID- 9854266 TI - How to study consciousness scientifically. AB - The neurosciences have advanced to the point that we can now treat consciousness as a scientific problem like any other. The problem is to explain how brain processes cause consciousness and how consciousness is realized in the brain. Progress is impeded by a number of philosophical mistakes, and the aim of this paper is to remove nine of those mistakes: (i) consciousness cannot be defined; (ii) consciousness is subjective but science is objective; (iii) brain processes cannot explain consciousness; (iv) the problem of 'qualia' should be set aside; (v) consciousness is epiphenomenal; (vi) consciousness has no evolutionary function; (vii) a causal account of consciousness is necessarily dualistic; (viii) science is reductionistic, so a scientific account of consciousness would show it reducible to something else; and (ix) an account of consciousness must be an information processing account. PMID- 9854267 TI - The spiritual variable in psychiatric research. PMID- 9854268 TI - Qualitative and quantitative analyses of a 'lock and key' hypothesis of depression. AB - BACKGROUND: We examine a 'lock and key' ('L-K') hypothesis to depression which posits that early adverse experiences establish locks that are activated by keys mirroring the earlier adverse experience to induce depression. METHODS: Two hundred and seventy clinically depressed patients were examined with open-ended and pre-coded interview questions to ascertain both early adverse experiences and precipitating life events. Qualitative and quantitative data analyses examined for any associations between developmental 'locks' and precipitating 'keys'. RESULTS: Qualitative assessment suggested 'L-K' links in almost one-third of the sample, and examples are provided. While quantitative analyses indicated significant associations between several identical 'lock' and 'key' constructs, evidence of specificity was rare. When individual 'locks' and 'keys' were consolidated into three higher-order constructs, variable models were suggested, including a non-specific link, a specific link and absence of any link. 'L-K' links appeared more likely in those with 'non-melancholic' (versus 'melancholic') depression, with the seemingly greater relevance to 'reactive' (versus 'neurotic') depression in the quantitative analyses inviting speculation that that 'disorder' may be more a reaction to a salient rather than a severe stressor. CONCLUSIONS: This exploratory study suggests that early adverse experiences may variably establish specific and non-specific patterns of vulnerability to having depression triggered by exposure to salient mirroring life event stressors. PMID- 9854269 TI - Life events and post-traumatic stress: the development of a new measure for children and adolescents. AB - BACKGROUND: A new interview measure of life events and post-traumatic stress disorder (PTSD) has been developed for children and adolescents aged 9 through 17, for use in both epidemiological and clinical studies. It includes 'high magnitude' events associated with PTSD as well as other 'low magnitude' events. METHOD: The interview is designed as a module of the Child and Adolescent Psychiatric Assessment, an interviewer-based interview conducted with parent and child separately by trained lay interviewers. The module includes: (1) questions about a wide range of events; (2) a screen for key PTSD symptoms (painful recall, avoidance, hypervigilance); and (3) a detailed interview on all PTSD symptoms, including onset, duration, severity and co-morbidity. A test-retest reliability study was conducted with 58 parents and children, who were interviewed twice by different interviewers. RESULTS: Intraclass correlations were 0.72 (child) and 0.83 (parent) for high magnitude events, and 0.62 (child) and 0.58 (parent) for low magnitude events. Kappa coefficients ranged from high for violence and sexual abuse to low for child reports of serious accidents and natural disasters. The reliability of the PTSD screen symptoms was fair to excellent (kappa = 0.40 0.79), and reliability of PTSD symptoms in those who passed the screen was excellent (ICC = 0.94-0.99). Compared with a general population sample (N = 1015), the clinic-referred subjects and their parents were twice as likely to report a traumatic event and, depending on the event, up to 25 times as likely to report symptoms of PTSD. CONCLUSIONS: The results support the reliability and discriminant validity of the measure. PMID- 9854270 TI - Which patients talk about stressful life events and social problems to the general practitioner? AB - BACKGROUND: Life events and social problems are common in primary-care attenders and contribute to the high rate of emotional distress observed in this setting but are often not disclosed during the consultation. Physicians' characteristics associated with patients' disclosure of psychosocial problems are well documented, but less empirical evidence is available on the contribution of patients' variables. METHOD: Logistic regression analyses were performed on a set of clinical and psychosocial data from 319 primary-care attenders with stressful life events and/or social problems in the presence or absence of emotional distress, in order to identify the variables that predict disclosure of such problems during the consultation. RESULTS: Two-thirds of patients with stressful life events and social problems had mentioned them to their GP. In both sexes a positive attitude about confiding and emotional distress were the best predictors of confiding. In men they accounted for 76% of correct predictions and, in women, together with past confiding, long-lasting relationship with GP, and coexistence of life events and social problems, for 81%. A positive confiding attitude was related in males to age and severity of medical condition and in females to age and experience of past confiding. CONCLUSIONS: In view of the high prevalence of positive attitude towards confiding the efforts by GPs should focus on the reduction of disclosure thresholds. This would require increased abilities to elicit psychosocial information and show emphatic understanding of patients' life difficulties. By contrast there is a need to promote a positive attitude in confiding that concerns only a minority of primary-care patients. More data are needed for shaping more specific strategies for these patients. PMID- 9854271 TI - Absence of social networks, social support and health services utilization. AB - BACKGROUND: This study examines the effects of distress and three types of social relationships (family, friends, and confidantes) on the use of out-patient health services in an adult sample from a community survey of 3481 persons in Baltimore, Maryland. METHODS: Independent effects of predisposing (age, education, marital status, race and sex), enabling (employment, income and insurance) and need (physical health) factors are adjusted for in estimating the odds of using health care services. RESULTS: Illness, being female, and having insurance were positively related to use of services, while being aged was inversely related. After adjustment for the above factors, social support interacted with psychological distress to effect the use of medical care: the combination of high distress and low social support by a confidante results in a fourfold increase of medical utilization. CONCLUSIONS: Our findings support the inclusion of psychological distress and social network variables in addition to physical health status in models attempting to explain the use of health services. Despite an inability to analyse change over time, our data suggest an understanding of the interrelationship between psychosocial factors, distress and health care use would benefit health providers and their patients. PMID- 9854272 TI - Symptoms of depression among community-dwelling elderly African-American and white older adults. AB - BACKGROUND: Few studies have explored the variance in individual symptoms by race in older adults. METHODS: Data were analysed from the Duke site of the Established Populations for Epidemiologic Studies of the Elderly (EPESE), a community sample of persons 65 years-of-age and older, 54% of whom were African Americans. Of the 3401 subjects with adequate data on depressive symptomatology, confirmatory factor analysis and LISREL were first used to confirm the presence of the factor structure previously reported for the CES-D. Next, bivariate analysis was performed to determine the prevalence of individual symptoms by race. Finally, LISREL analysis was performed to control for potential confounding variables. RESULTS: When bivariate comparisons of specific symptoms by race were explored, African-Americans were more likely to report less hope about the future, poor appetite, difficulty concentrating, requiring more effort for usual activities, less talking, feeling people were unfriendly, feeling disliked by others and being more 'bothered' than usual. When LISREL analyses were applied to these data (controlling for education, income, cognitive impairment, chronic health problems and disability and other factors) racial differences in somatic complaints and life satisfaction disappeared, yet differences in interpersonal relations persisted. CONCLUSIONS: This study confirms earlier findings of minimal overall differences in symptom frequency between African-American and non-African American community-dwelling older adults in controlled studies. PMID- 9854273 TI - Symptoms of depression and anxiety during adult life: evidence for a decline in prevalence with age. AB - BACKGROUND: To test the hypothesis that the prevalence, in the general population, of symptoms of depression and anxiety declines with age. METHODS: A general population sample of 2725 persons aged 18 to 79 years was administered two inventories for current symptoms of depression and anxiety, together with measures of neuroticism and of exposures that may confer increased risk of such symptoms. RESULTS: Symptoms of depression showed a decline with age in both men and women. For anxiety, the decline was statistically significant for women but not consistently so for men. For the risk factors examined, there was a decline with age in the neuroticism score, the frequency of adverse life events, being seriously short of money and having had parents who separated or divorced. Further analysis showed that the association between age and a declining symptom score cannot be entirely attributed to these risk factors, with the single exception of neuroticism. The latter is itself likely to be contaminated by current symptoms. CONCLUSION: Unless these findings are due to bias in the sample of those who agreed to participate, they add to the evidence that symptoms of depression and to a lesser extent of anxiety decline in prevalence with age. Some risk factors also decline with age. It now has to be determined if these cross sectional observations are also to be found in longitudinal data; and what process may underlie this striking change in mental health during adulthood. PMID- 9854274 TI - Outcome of the depressed elderly living in the community in Liverpool: a 5-year follow-up. AB - BACKGROUND: Comparatively little is known about the long-term natural history of depressive disorders in the elderly living in the community. This is a follow-up of a subsample of the Continuing Health in the Community study random sample of the elderly population living in Liverpool. METHODS: The investigators followed up 120 cases of depression identified by a semi-structured interview schedule (GMS) for a period of 5 years. A similar number of other subjects defined as subcases of depression, other cases of mental illness and a random selection of non-cases were also included. RESULTS: The 5-year outcome for the cases of depression was worse than the outcome of the non-cases (relative mortality risk of 2.1, 95% confidence interval 1.1 to 3.9). Thirty-four per cent of the cases of depression died and 28% had dropped out during the follow-up. Of the 46 cases of depression who had a complete follow-up, 22% recovered from their symptoms, 30% were found to be AGECAT cases at one of the three follow-up waves, 24% were AGECAT cases at two of the three follow-up waves and the remaining 24% were AGECAT cases at each follow-up wave. Fifteen per cent of the surviving cases of depression were organic cases at the follow-up. Their anxiety comorbid state and depression score were identified as predictors of poor outcome. CONCLUSION: The findings of this study indicate that depressive disorders (most of which were untreated) found in the elderly community have a poor prognosis. PMID- 9854275 TI - Prevalence of depression in an elderly community sample: a comparison of GMS AGECAT and DSM-IV diagnostic criteria. AB - BACKGROUND: Prevalence rates of depression based on the GMS-AGECAT system are markedly higher than rates of major depression diagnosed using DSM-III and DSM III-R criteria. Interpreting this finding is difficult since AGECAT diagnostic criteria have not been published. METHODS: We conducted a survey in Edmonton, Canada, in which 1119 community residents aged 65 years or older were administered the Geriatric Mental State (GMS) questionnaire. Depression was diagnosed using AGECAT and also according to criteria for DSM-IV major depression and minor depression. We identified 57 items in the GMS relating to major and minor depression which were then grouped into dysphoric, somatic and cognitive categories. The relationship of these symptoms to AGECAT depression was investigated by recording symptoms to absent (nullifying) and recalculating the number of AGECAT cases. Data were weighted to reflect the underlying population. RESULTS: There were 143 cases of AGECAT depression (prevalence rate = 11.4%), 14 cases of major depression (prevalence rate = 0.86%) and 44 cases of minor depression (prevalence rate = 3.6%). Of the 154 subjects with either AGECAT or DSM-IV depression, only 47 (31%) were depressed according to both diagnostic systems. Nullifying all dysphoric symptoms reduced the number of AGECAT cases to eight, whereas nullifying somatic and cognitive symptoms reduced the numbers of cases only to 138 and 142, respectively. By themselves, dysphoric symptoms accounted for 77 of the 143 cases of AGECAT depression, while somatic and cognitive symtpoms alone accounted for no cases. CONCLUSIONS: AGECAT depression is more inclusive than DSM-IV major or minor depression and AGECAT case status is determined mainly by the proportion of dysphoric symptoms. PMID- 9854276 TI - A randomized controlled trial of guided imagery in bulimia nervosa. AB - BACKGROUND: The objective of this study was to test a guided imagery therapy designed to enhance self-comforting in bulimia nervosa. METHODS: A randomized controlled trial compared 6 weeks of individual guided imagery therapy with a control group. Fifty participants who met DSM-III-R criteria for bulimia nervosa completed the study. Measures of eating disorder symptoms, psychological functioning and the experience of guided imagery therapy were administered. RESULTS: The guided imagery treatment had substantial effects on the reduction of bingeing and purging episodes; the imagery group had a mean reduction of binges of 74% and of vomiting of 73%. The imagery treatment also demonstrated improvement on measures of attitudes concerning eating, dieting and body weight in comparison to the control group. In addition, the guided imagery demonstrated improvement on psychological measures of aloneness and the ability for self comforting. CONCLUSIONS: Evidence from this study suggests that guided imagery was an effective treatment for bulimia nervosa, at least in the short-term. PMID- 9854277 TI - Randomized trial of physical exercise alone or combined with bright light on mood and health-related quality of life. AB - BACKGROUND: So-called atypical depressive symptoms (carbohydrate craving, prolonged sleep, weight gain, increased appetite) frequently emerge in association with low illumination to which people are ordinarily exposed indoors, or even outdoors at extreme latitudes in wintertime. Our objective was to analyse the effect of physical exercise alone or combined with bright light on mood and the health-related quality of life during winter. METHODS: We carried out a randomized controlled trial on 120 indoor employees in southern Finland between November and January. The subjects were allocated to supervised fitness training under bright (2500-4000 lx) or ordinary (400-600 lx) light conditions in a gym 2 3 times weekly for 8 weeks, or supervised relaxation training once a week over the same period as active placebo. We collected questionnaire data on the changes in mood and health-related quality of life after 4 and 8 weeks of training, and after 4 months follow-up. RESULTS: Fitness training in bright light resulted in greater relief from atypical depressive symptoms and more vitality than in ordinary room light. Compared with relaxation alone, the former regime improved general mental health and social functioning in addition to the improvement in depressive symptoms and vitality, whereas the latter only increased vitality. CONCLUSIONS: Supervised physical exercise combined with exposure to bright light appears to be an effective intervention for improving mood and certain aspects of the health-related quality of life in wintertime. This effect appears unrelated to the history of season-dependent symptoms, being noticeable among healthy individuals. PMID- 9854278 TI - A four year prospective study of age-related cognitive change in adults with Down's syndrome. AB - BACKGROUND: While neuropathological studies indicate a high risk for Alzheimer's disease in adults with Down's syndrome, neuropsychological studies suggest a lower prevalence of dementia. In this study, cognitive deterioration in adults with Down's syndrome was examined prospectively over 4 years to establish rates and profiles of cognitive deterioration. METHODS: Fifty-seven people with Down's syndrome aged 30 years or older were assessed using a battery of neuropsychological tests on five occasions across 50 months. Assessments of domains of cognitive function known to change with the onset of Alzheimer related dementia were employed. These included tests of learning, memory, orientation, agnosia, apraxia and aphasia. The individual growth trajectory methodology was used to analyse change over time. RESULTS: Severe cognitive deterioration, such as acquired, apraxia and agnosia, was evident in 28.3% of those aged over 30 and a higher prevalence of these impairments was associated with older age. The rate of cognitive deterioration also increased with age and degree of pre-existing cognitive impairment. Additionally, deterioration in memory, learning and orientation preceded the acquisition of aphasia, agnosia and apraxia. CONCLUSIONS: The prevalence of cognitive impairments consistent with the presence of Alzheimer's disease is lower than that suggested by neuropathological studies. The pattern of the acquisition of cognitive impairments in adults with Down's syndrome is similar to that seen in individuals with Alzheimer's disease who do not have Down's syndrome. PMID- 9854279 TI - Homeless youth in London: I. Childhood antecedents and psychiatric disorder. AB - BACKGROUND: There has been an increase in the numbers of homeless young people in Britain. Little is known of the health and social welfare needs of this population. METHOD: This case-control study compares a random sample of homeless people aged under 22 years recruited from consecutive attenders at two of London's largest facilities for homeless young people with a contemporaneous sample of domiciled young people recruited through general practice registration lists. The homeless and domiciled groups were compared on measures of childhood care, education and psychiatric disorder. RESULTS: One hundred and sixty-one homeless people (88% of those approached) and 107 domiciled subjects (60% of those approached) were interviewed. Sixty-nine per cent of homeless and a third of the domiciled subjects reported a childhood lacking in affection, with indifferent and often violent carers. Psychiatric disorder was identified in 62% of homeless respondents and a quarter of the domiciled population. A fifth of homeless and 5 domiciled respondents had attempted suicide in the previous year. Multivariate analysis suggest that childhood adversity, low educational attainment and the prior presence of psychiatric disorder all independently increase the likelihood of homelessness in a youthful population. CONCLUSIONS: The evidence presented in this paper supports the hypotheses that characterize the young homeless population as experiencing higher rates of childhood adversity and psychiatric disorder than their domiciled contemporaries. A tentative model is suggested whereby childhood experiences, educational attainment and the prior presence of psychiatric disorder all independently increase the likelihood of homelessness in a youthful population. PMID- 9854280 TI - Assortative mating for major psychiatric diagnoses in two population-based samples. AB - BACKGROUND: Previous studies on assortment for psychiatric disorders have reported discrepant findings. We aimed to test whether there is a significant association for psychiatric diagnoses, including alcoholism, generalized anxiety disorder, major depressive disorder, panic disorder and phobias between husbands and wives in two population-based samples. We further evaluated whether marital resemblance occurs primarily within or across psychiatric disorders and if assortment for psychopathology is primary or secondary to assortment for correlated variables. METHODS: A model for mate selection addressed whether the correlation between mates for psychiatric disorders arises from direct assortment (primary homogamy) or through correlation with other variables for which assortment occurs (secondary homogamy) or through cross-variable assortment. The model accounted for within-person co-morbidity as well as across-spouse data. RESULTS: Findings suggested that a moderate degree of assortment exists both within and across psychiatric diagnoses. Only a small amount of the observed marital resemblance for mental illness could be explained by assortment for correlated variables such as age, religious attendance and education. Similar results were obtained for the two samples separately and confirmed in their joint analysis, revealing that the co-morbidity and assortment findings, except for the marital correlation for age, religious attendance and education, replicate across samples. CONCLUSIONS: Significant but moderate primary assortment exists for psychiatric disorders. The bias in twin studies that have ignored the small amount of assortment is negligible. PMID- 9854281 TI - A population-based twin study of self-esteem and gender. AB - BACKGROUND: Self-esteem (SE), a widely used construct in the social sciences, is usually conceptualized as a reflection of socialization and interpersonal experiences that may differ considerably between the genders. METHODS: The Rosenberg self-esteem scale was assessed at personal interview in both members of 3793 unselected twin pairs (1517 male-male, 856 female-female and 1420 male female) from the population-based Virginia Twin Registry. Gender effects on SE were assessed by both analysis of variance and biometrical twin modelling. RESULTS: The mean SE score was slightly but significantly lower in women v. men, and in women who grew up with a male v. a female co-twin. Twin modelling suggested that: (i) individual differences in self-esteem in both men and women were best explained by genetic and individual-specific environment factors; (ii) heritability estimates were similar in women (32%) and in men (29%); and (iii) the same genetic factors that influenced SE in women also influenced SE in men. Analyses supported the validity of the equal environment assumption for SE. The heritability of SE cannot be explained by the moderate correlation between SE and symptoms of depression. CONCLUSIONS: These results are inconsistent with prominent gender-related aetiological models for SE, which postulate that individual differences arise from socialization experiences both within and outside the home of origin which differ widely for the two genders. Instead, a significant proportion of the population variance in SE is due to genetically influenced temperamental variables that are the same in men and women. PMID- 9854282 TI - Prevalence and severity of substance use disorders and onset of psychosis in first-admission psychotic patients. AB - BACKGROUND: Past studies have found inconsistent evidence that substance use disorders are related to earlier onset of schizophrenia or more severe symptoms. This study examines prevalence and severity of current substance use disorders and onset of psychotic illness in a multi-facility sample. METHODS: Data are from the Suffolk County Mental Health Project, an epidemiological study of first admission psychosis. The SCID and instruments measuring symptomatology, personality and background characteristics were administered. Respondents were stratified into three groups: (a) no life-time substance diagnosis; (b) in remission or reporting current mild use at admission; and (c) current moderate severe substance abuse at admission. RESULTS: Using the SCID severity rating, 17.4% of males and 6.2% of the females had moderate or severe current substance abuse, while 41.5% of males and 68.2% of females had no lifetime substance diagnosis. In almost all cases categorized as moderate-severe, the substance diagnosis predated onset of psychosis. Females categorized as moderate-severe have an earlier age of onset of psychosis than did females in the other groups. There were only slight differences in symptom severity among the groups but more marked antisocial behaviour in the moderate-severe group. Variables discriminating the moderate-severe from non-abuse groups were BPRS thought disturbance, adult anti-social behaviour and current cigarette smoking for males and adult anti-social behaviour and child-teen antisocial behaviour for females. CONCLUSIONS: Severity of substance abuse does not appear to be a pivotal correlate of the early features of psychotic illness. PMID- 9854283 TI - Multivariate analysis of outcome of mental health care using graphical chain models. The South-Verona Outcome Project 1. AB - BACKGROUND: Short-term outcome of mental health care was assessed in a multidimensional perspective using graphical chain models, a new multivariate method that analyses the relationship between variables conditionally, i.e. taking into account the effect of antecedent and intervening variables. METHODS: GAF, BPRS, DAS (at baseline and after 6 months), LQL and VSSS (at follow-up only) were administered to 194 patients attending the South-Verona community-based mental health service. Direct costs in the interval were also calculated. Graphical chain models were used to analyse: (1) the associations between predictors (psychopathology, disability, functioning, assessed at baseline); (2) the effects of predictors on costs; and (3) the effect of predictors and costs on outcomes (psychopathology, disability, functioning, quality of life and service satisfaction) as well as their correlation. RESULTS: Psychopathology, disability and functioning scores at baseline predicted the corresponding scores at 6-month follow-up, with greater improvement in the more severely ill. Higher psychopathology and poorer functioning at baseline predicted higher costs and, in turn, costs predicted poorer functioning at follow-up. Outcome indicators polarized in two groups: psychopathology, disability and functioning, which were highly correlated; and the dyad service satisfaction and quality of life. Service satisfaction was highly related to quality of life and was predicted by low disability and high dysfunctioning. No predictors for quality of life were found. CONCLUSIONS: Graphical chain models were demonstrated to be a useful methodology to analyse process and outcome data. The results of the present study help in formulating specific hypotheses for future studies on outcome. PMID- 9854284 TI - Aspects of dysphoria and symptoms of schizophrenia. AB - BACKGROUND: In the past it has been postulated that dysphoric emotions may be related to positive and/or negative symptoms in schizophrenia. The results of several recent studies have suggested that composite dysphoria indices are more strongly related to positive than negative symptoms. In the current study we use part correlation techniques to examine the possible unique contributions of two aspects of dysphoria--depression and anxiety--to three syndromes of symptoms (reality distortion, disorganization and psychomotor poverty) within schizophrenia. METHODS: Data were obtained from 60 patients with a DSM-III-R diagnosis of schizophrenia. Symptoms of schizophrenia were assessed using the SAPS and SANS and dysphoria was assessed using both self-report (BDI and BAI) and observer ratings (HRSD and HARS). Assessment of schizophrenia symptoms and ratings of depression and anxiety were completed by different observers. In addition, drug induced extrapyramidal side effects were rated. RESULTS: Part correlations showed that unique aspects of anxiety (particularly physiological arousal) were correlated with reality distortion while unique aspects of depression (including psychomotor slowing and loss of social interest) were related to psychomotor poverty. At least part of the latter relationship may be due to extrapyramidal side effects of neuroleptic medication. CONCLUSIONS: Although there is considerable overlap between anxiety and depression, it appears that the unique arousing or activating aspects of anxiety are related to the experience of reality distortion symptoms in schizophrenia and the unique slowing and withdrawal aspects of depression are particularly related to psychomotor poverty. Possible reasons for these relationships are discussed. PMID- 9854285 TI - Recollections of parental behaviour, adult attachment and mental health: mediating and moderating effects. AB - BACKGROUND: Attachment theory posits links between early experiences with parents, adult relationships and adult mental health, but does not specify whether these are independent, mediating, or moderating effects. METHODS: Associations of parent's behaviour on the Parental Bonding Instrument, adult attachment styles and three dimensions of mental health were investigated in a large sample of women and men. RESULTS: Men and women with secure styles recalled higher levels of care from both parents than those with fearful styles. Maternal and paternal control were more consistent predictors of increased distress for men than for women. Fearful and preoccupied adult styles were associated with higher levels of distress in both men and women. While adult styles had few mediating effects on the association of parental behaviour and mental health, interactions between the fearful style and parental variables suggested that this form of insecurity sometimes accentuated the impact of high parental care or low paternal control on mental health in both men and women; among women, however, the secure style seemed to buffer somewhat the negative effect of high parental control. CONCLUSION: Although the amount of variance explained by either parental behaviour or adult styles was modest, patterns of moderating effects of adult styles on associations between parental behaviour and mental health suggested that both continuity and discontinuity principles can be applied to understanding these links. PMID- 9854286 TI - Is introversion a risk factor for suicidal behaviour in depression? AB - BACKGROUND: Personality is an important determinant of suicidal behaviour. However, it has been studied little in relation to suicidal behaviour in depression. METHODS: Depressed patients who had attempted suicide (N = 41) were compared with depressed patients who had never attempted suicide (N = 56) and normal controls (N = 56) for their scores on three personality questionnaires. RESULTS: Introversion was the only personality dimension where the post hoc test showed a different pattern between the two groups of depressed patients in their relation to controls. In particular, only depressed patients who had attempted suicide had significantly lower introversion scores than controls. CONCLUSION: The personality dimension of introversion may be a risk factor for suicidal behaviour in depression. PMID- 9854287 TI - General practitioners' attitudes to psychiatric and medical illness. AB - BACKGROUND: General practitioners are increasingly involved in the care of patients with long-term psychiatric disorders. We have previously reported that general practitioners are less willing to treat patients with schizophrenia than those without such a diagnosis, but this may have been attributable to a reluctance to treat patients with any psychiatric or chronic illness. We, therefore, examined general practitioners' attitudes to patients with chronic psychiatric or medical illnesses. METHODS: A random sample of 260 local general practitioners were each sent one of our case vignettes which were identical apart from mention of a previous diagnosis of schizophrenia, depression, diabetes or no illness. The general practitioners were asked to indicate their level of agreement with 13 attitudinal statements based on the vignette. RESULTS: One hundred and sixty-six (66%) of the general practitioners responded to the case vignettes. Those responding to the vignette about the patient with schizophrenia were less happy to have that patient on their practice list and were more concerned about the risk of violence and the child's welfare. Those responding to the depression vignette were more likely to offer the patient antidepressants or counselling; and those who replied to the diabetes case were most likely to refer the patient to a hospital specialist. These differences were not attributable to the personal or practice characteristics of the general practitioners. CONCLUSIONS: Patients with schizophrenia arouse concerns in general practitioners that are not simply due to those patients suffering from a psychiatric or chronic illness. Our results suggest that some patients with schizophrenia may find it difficult to register with a general practitioner and receive the integrated community-based health care service they require. Psychiatrists should provide education and support to general practitioners who look after patients with schizophrenia. PMID- 9854288 TI - [Megacolon and megarectum in adults. Abdominal proctectomy with colo-anal anastomosis]. AB - Severe constipation in adults with megarectum may be a manifestation of Hirschsprung's disease or idiopathic megarectum. The differential diagnosis is discussed in the light of two case histories. Total abdominal proctectomy with coloanal anastomosis eliminated the aganglionic rectal segment in Hirschsprung's disease and the adynamic distal colorectum of idiopathic megarectum respectively. The surgical and functional results of a hand-sewn transanal end-to-end anastomosis were excellent in both patients. PMID- 9854289 TI - [Vitamin B 12 deficiency: early diagnosis in ambulatory care medicine]. AB - Many patients suffer from vitamin B12 deficiency and are thus exposed to irreversible sequelae if diagnosis occurs at a late stage. This prospective study undertaken by eight practitioners over a period of 12 months concerns early diagnosis. Blood vitamin B12 levels were measured in 152 patients presenting macrocytosis detected by systematic MCV analysis at the time of a blood test, a neuropathy or a recent cognitive, affective and behavioural problem, and were found to be lowered (< or = 175 pmol/l) in 54 patients of whom 43 had undergone vitamin B12 test treatment for 6 months. Haematological, neurological and psychiatric evaluation was carried out before and after treatment, and a diagnosis of deficiency was recorded in 24 patients based on unequivocal response to therapy. Improvement was greatest haematologically in 12 patients, neurologically in 6 patients and psychiatrically in 6 other patients, with 4 patients showing a combination of all modes. These 24 patients (mean age 69 years) suffered from numerous pathologies which were liable to complicate diagnosis in some of them: neurological (46%), psychiatric (37%), chronic alcoholism (33%), folic acid deficiency (29%), and diabetes (17%). The only diagnostic element used as a criterion of deficiency was an extremely low level of vitamin B12 (< or = 75 pmol/l). Marked macrocytosis or a combination of haematological and neuropsychiatric signs are strong indicators, but only improvement under treatment allowed a diagnosis to be made in the majority of patients. Macrocytosis was, however, not present in 6 of the 12 neuropsychiatric patients. The study thus identified a high proportion of patients with vitamin B12 deficiency who additionally presented, in equal proportions, both haematological and neuropsychiatric symptoms. Neither the clinical examination nor the vitamin B12 level in general permit early diagnosis based on a high probability index and long-term follow-up. Simpler methods for early diagnosis are therefore needed. PMID- 9854290 TI - [Preoperative assessment of primary varicose veins]. AB - The first step in the evaluation of a patient with primary varicose veins is a careful personal history and clinical examination. The individual advice should be based on the disturbance suffered due to the actual varicose veins, "heaviness of the legs" and the advantages and disadvantages of various therapies. The aim of an examination in any type of varicose veins is to determine their relationship to the deep and the surrounding superficial venous system. The continuous-wave Doppler method is often sufficient to examine the trunk of the long saphenous vein. Colour-coded duplex sonography is the chosen method for evaluation of the deep venous system with respect to flow feasibility and valve competence, as well as localization of deficient perforants and anatomic variations at the saphenouspopliteal junction. Phlebography is reserved for cases with inconclusive duplex-sonography results or if no duplex is available. The purpose of this exhaustive preoperative evaluation is to indicate in detail the haemodynamic and anatomic situation in the groin and in the popliteal fossa and to detect the relevant insufficient perforators. Such accurate haemodynamic assessment is the basis for an individualized operation with excellent cosmetic and functional results. Whether a reduced recurrence rate after varicose vein surgery can be shown with this extensive examination scheme has not yet been investigated in a prospective study. PMID- 9854291 TI - [Surgical therapy of primary varicose veins]. AB - The principle of varicose vein surgery still remains the interruption of all insufficient communications between the deep and the superficial venous system and removal of the varicosities. The basis for differentiated surgical treatment is accurate preoperative assessment. Careful dissection of the saphenofemoral junction through a suprainguinal incision, with division of all the branches and flush tie of the long saphenous vein combined with invaginated stripping of the long saphenous vein to just below the knee, appears to be the method of choice for good clinical results and a low incidence of damage to the saphenous nerve. Oesch recently introduced a new technique of perforate invaginate (PIN) stripping which gives even better cosmetic results. Regarding the short saphenous vein, preoperative localization of the exact level of the saphenopopliteal junction is of major importance in the prevention of recurrence. Simple evulsion or epifascial or subfascial ligation were the most common treatments for incompetent perforating veins for many years. In 1985 Hauer described endoscopic subfascial dissection of perforating veins (ESDP), which reduces delayed wound healing, especially in trophic skin changes. Deprivation of blood supply with a pneumatic tourniquet such as the Lofqvist roller cuff is necessary. The tributaries are removed by stab evulsion phlebectomy with specially designed hooks. This technique was originally introduced by Muller for ambulatory treatment of varicose veins. The incisions of 1-3 mm guarantee excellent cosmesis and minimal trauma. Adhesive tape is used to close the incisions. A number of alternative techniques such as cryosurgery, laser surgery, paratibial fasciotomy and the CHIVA technique (Conservative Treatment and Haemodynamics in Venous Insufficiency in Outpatient Departments) are briefly described. Complications of varicose vein surgery are rare. Minor complications are skin nerve injuries, haematomas, infections and lymphatic fistulas. Major complications such as injuries to the femoral vein or artery occur in less than 0.05%. But once it has occurred it is of paramount importance to recognize the injury at the time of initial surgery, to avoid limb loss. Provided the preoperative assessment is accurate and the principles of selective surgical treatment are followed, the surgeon is able to perform a curative operation with a low complication rate and excellent cosmetic results. PMID- 9854292 TI - [Red fingers]. PMID- 9854293 TI - Medical schools in the new millenium. A reply. PMID- 9854294 TI - A melange of meningiomas (Scot Med J 1998; 43: 117-118) PMID- 9854295 TI - Demise of mercury manometer. PMID- 9854296 TI - Falls in old age: inevitable or preventable? PMID- 9854297 TI - Associations between road traffic accidents and socio-economic deprivation on Scotland's west coast. AB - Road traffic accidents (RTAs) are declining, but remain a public health concern locally and world-wide. Scottish RTAs killed 316 people and injured over 20,000 in 1996. By 2020, they are predicted to become the world's third-leading cause of sickness and death. Little is know about associations between RTAs and deprivation; it has never been explored on Scotland's West Coast. This study analysed hospital A&E admissions and investigated associations between RTAs and socio-economic status. 1,300 attendance records at a 575-bed NHS Trust Accident & Emergency in North Lanarkshire were reviewed and 1,020 records analysed in conjunction with Health Board socio-economic data. Findings strongly suggest (p = 0.00461) a positive trend between RTA activity and deprivation. Significance held for gender, victim role, purpose of journey and age, except for drivers 60 and over. Given the preventative nature of RTAs and their contribution to morbidity and mortality, further research between RTAs and deprivation is suggested. PMID- 9854298 TI - Golf related head injuries in children: the little tigers. AB - Head injury is a major cause of morbidity in Western society and sport related incidents account for approximately 11% of all head injured patients attending Accident and Emergency Departments. Golf was shown to be one of the sports most commonly associated with head injury requiring referral to a regional neurosurgical centre. Previous studies have demonstrated that it is predominantly children who sustain golf related head injuries which present either to an accident and emergency department or a regional neurosurgical centre. This study examines the number and pattern of golf related head injuries in children presenting to an accident and emergency department or requiring admission to the regional neurosurgical centre, over a three month period. PMID- 9854299 TI - Plastic surgery waiting list--the numbers game fact or fiction? AB - Previous studies have indicated the importance of work load and operating time in helping to manage a waiting list. While analysis of a waiting list based solely on numbers cannot give an accurate assessment it is only this and "the waiting time" to surgery (apparently irrespective of clinical priorities) that seem to matter to the government, the Trust Hospitals, and to the public. This study examines case numbers on a waiting list over a two year period and links this to the preceding six months elective operations. Further insight into a waiting list construction can be obtained because of the clinical need for some degree of prioritisation, particularly because of malignancies, it is obvious why the bare waiting list numbers and composition can give a false impression of the true situation. The secondary effect on waiting time for less urgent cases can thus be more readily explained. Some form of rationing may be the answer. PMID- 9854300 TI - Variations in the management of singleton breech presentation throughout Scotland. AB - Breech presentation occurs in 3-5% of deliveries and can be managed by either a trial of vaginal breech delivery (TOVBD), external cephalic version (ECV) or Caesarean section. A postal questionnaire was completed by 82% of Scottish consultant obstetricians and revealed wide variations in practice. Eighteen percent never offered ECV. Among those who did consensus was lacking on some contraindications. One-quarter sometimes performed ECV before 37 weeks gestations despite the possibility of spontaneous version. Only 70% restricted ECV to one or more designated operators thereby maintaining levels of expertise. Variations were demonstrated in the use of tocolytics, and pre and post procedure investigations. Following failed ECV 28% considered a repeat attempt and 56% a TOVBD. TOVBD was not offered as first line management by one-fifth of respondents. Those who did varied in the pre-procedure investigations performed. Guidelines are required to ensure safe, consistent practice and avoid unnecessary Caesarean sections. PMID- 9854301 TI - Raw egg ingestion and salmonellosis in body builders. AB - Four patients with Salmonella enteritidis infection are reported. All were body builders who regularly consumed substantial quantities of raw eggs. They presented with a severe febrile illness and diarrhoea--presumably reflecting a large bacterial inoculum. Advice regarding the potential hazards of raw egg ingestion has been repeatedly issued by the Department of Health--but this report highlights the fact that this practice continues in spite of this. The epidemiology of S. enteritidis infection in relation to raw egg ingestion is discussed. PMID- 9854302 TI - Neisseria meningitidis W135 pneumonia with sepicaemia in a nonogenarian. AB - Neisseria meningitidis infection is generally considered a disease of children or young adults, classically presenting as meningitis or sepicaemia. This infection is rare but recognised in the elderly. We present the case of a nonogenarian with meningococcal pneumonia and sinusitis with bacteraemia caused by N.meningitidis W135 a rare serogroup. We therefore thought this unusual situation of interest and worthwhile reporting. PMID- 9854303 TI - Hypoplastic anaemia complicating myxoedema coma. AB - The case of a 68 year old women presenting in myxoedema coma is described. She was found to be anaemic with a haemoglobin of 8.2 g/dl. Further investigations showed a pancytopenia and a hypoplastic anaemia confirmed by bone marrow. The patient recovered and became euthyroid following initial treatment with intravenous tri-iodothyronine (T3) and later oral thyroxine (T4) replacement with resolution of pancytopenia and return of bone marrow to normal. PMID- 9854304 TI - The use of audit to improve clinical effectiveness in an infrequent obstetric therapy: magnesium sulphate in severe pre-eclampsia. AB - Magnesium sulphate has been established as the drug of choice in the management of eclampsia and even when used in cases of severe pre-eclampsia it is given infrequently. It is recognised to have potentially severe toxic effects but there is a need to achieve therapeutic levels quickly enough to prevent seizures. This paper reports an audit following the introduction of a protocol for the use of magnesium sulphate in eclampsia and severe pre-eclampsia at Ninewells hospital, Dundee. Problems were identified with both the time taken to reach therapeutic levels and clinical monitoring of magnesium levels despite the use of loading doses commonly recommended. Reaudit after changes to the protocol and the introduction of an administration/monitoring chart showed a significant improvement in both monitoring and the adequacy of prophylaxis. This demonstrates how audit of clinical practice has been used to improve clinical effectiveness in an area in which a potentially toxic drug is used infrequently but with potentially life saving benefits. PMID- 9854305 TI - Blood ordering practices in elective and emergency surgical procedures. PMID- 9854306 TI - Alexander Ogston and the Army Medical Services formation of the Royal Army Medical Corps 1 July 1898. AB - The Royal Army Medical Corps came into existence on 1st July 1898, in response to dissatisfaction with the organisation and efficiency of the Army Medical Service. Although much of the pressure for change came from officers within the service, a significant factor was the campaign conducted by a civilian, Alexander Ogston, Professor of Surgery at Aberdeen. PMID- 9854307 TI - Tau in Alzheimer's disease. AB - Neurofibrillar protein aggregates containing tau are one of the major hallmarks of Alzheimer's disease (AD). In normal cells, tau stabilizes axonal microtubules, which are the tracks for intracellular traffic. In AD, tau becomes abnormally phosphorylated, aggregates into paired helical filaments and loses its ability to maintain the microtubule tracks. There is renewed interest in tau as a causative factor in neurodegenerative disease based on recently discovered mutations in the gene encoding tau. This article discusses how changes in tau protein could lead to retraction of neuronal processes and thus cell death and argues that tau pathology, rather than beta-amyloid, might be the most reliable indicative factor for AD. PMID- 9854308 TI - C1q--how many functions? How many receptors? AB - C1, the first component of the classical pathway of complement activation is a complex of three proteins called C1q, C1r and C1s. Normally, C1q binding to aggregated IgG molecules results in activation of the classical pathway of complement. However, C1q has a number of other observed functions, not directly related to complement, that could be mediated by recently identified binding proteins acting as cell-surface receptors or soluble modulators of C1q-mediated functions. This article discusses the various activities of C1q and the evidence that these functions might be influenced by both membrane-bound and soluble C1q binding proteins. PMID- 9854309 TI - Pictures in cell biology. Un-LOXing the door to organelle degradation. PMID- 9854310 TI - ECM signalling: orchestrating cell behaviour and misbehaviour. AB - The extracellular matrix (ECM) provides signalling cues that regulate cell behaviour and orchestrate functions of cells in tissue formation and homeostasis. The composition of the ECM, its three-dimensional organization and proteolytic remodelling are major determinants of the microenvironmental signalling context that controls cell shape, motility, growth, survival and differentiation. In recent years, the importance of ECM signalling has been underscored by the evidence that misregulation of cell-ECM interactions can contribute to many diseases, including developmental, immune, haemostasis, degenerative and malignant disorders. This review discusses recent insights into the regulatory functions of the ECM and their role in disease, focusing on cancer as a paradigm. PMID- 9854312 TI - The cell biology of beta-amyloid precursor protein and presenilin in Alzheimer's disease. AB - It is a truism of modern biomedical science that the development of therapies expected to slow or arrest the progression of a disease requires as detailed an understanding of its molecular and cellular pathogenesis as possible. In turn, the cloning of novel gene products implicated in a disease often leads to new insights about fundamental features of protein structure and function. A particularly compelling example of this beneficial interplay between basic and applied cell biology arises from the exciting recent progress in deciphering Alzheimer's disease (AD). This review discusses the current understanding of the cell biology of two proteins crucial for the pathogenesis of AD, the beta-amyloid precursor protein and presenilin. PMID- 9854311 TI - Direct targets of phosphoinositide 3-kinase products in membrane traffic and signal transduction. AB - Phosphoinositide 3-kinases are a family of lipid kinases that phosphorylate the 3' position of the inositol ring on phosphatidylinositol and higher phosphorylated polyphosphoinositides. The multiplicity of cellular functions influenced by the activity of these enzymes has captured the attention of researchers working on two important fields of cell biology--signal transduction and membrane traffic. This review discusses how the recent identification of proteins that interact directly with 3'-polyphosphoinositides has revealed novel potential interconnections between these fundamental cellular processes. PMID- 9854313 TI - SMC proteins and chromosome structure. AB - The structure of chromosomes is largely determined by chromosome-associated proteins. Members of the SMC (structural maintenance of chromosomes) family play an important role in both prokaryotic and eukaryotic chromosome structure and dynamics. SMC proteins are involved in chromosome condensation, sister-chromatid cohesion, sex-chromosome dosage compensation, genetic recombination and DNA repair. There have been major advances recently in understanding the function of SMC proteins--including the identification of biochemical activities of SMC containing protein complexes and the realization that individual SMC proteins might link seemingly unrelated aspects of chromosomal metabolism. PMID- 9854314 TI - Daniel Branton and freeze-fracture analysis of membranes. PMID- 9854315 TI - Mastitis therapy and milking machines: the 1998 British Mastitis Conference. PMID- 9854316 TI - Paralysis in hedgehogs (Erinaceus europaeus) associated with demyelination. AB - Paraplegia affected 14 hedgehogs (Erinaceus europaeus) in a wildlife rescue hospital over a period of six months. Postmortem examination revealed demyelination in the brain and spinal cord and an inflammatory response in the meninges, choroid plexus and CNS. The peripheral nervous system was not affected. In the spleen, lungs and liver there was an accumulation of megakaryocytes and other evidence of extramedullary haemopoiesis, but there was no haematological evidence of anaemia. The pattern of disease incidence and the nature of the changes in the CNS suggest they were of viral origin, but no causal agent was isolated and the possibility of a neurotoxin cause cannot be ruled out. PMID- 9854317 TI - Persistence of Ehrlichia phagocytophila infection in lambs in relation to clinical parameters and antibody responses. AB - Five lambs were infected experimentally with Ehrlichia phagocytophila and examined regularly during the next six months. The lambs all had recurrences of parasitaemia at various times but had a fever on only 21 per cent of these occasions. A reduced number of leucocytes was observed in all the lambs for at least eight weeks. All the lambs were still infected four months after inoculation with E phagocytophila. After six months, blood from four of the five lambs was infective when inoculated into susceptible lambs. PMID- 9854318 TI - Surgical removal of an undifferentiated abdominal sarcoma from a koi carp (Cyprinus carpio). AB - An undifferentiated abdominal sarcoma was removed surgically from a koi carp. The diagnostic procedures, including radiography and computed tomography, and the procedures for general anaesthesia and the surgical approach for a celiotomy in a fish are described. The gross and microscopic appearance of the tumour is described and illustrated. PMID- 9854319 TI - Efficacy of a 2 per cent moxidectin gel against gastrointestinal parasites of ponies. AB - The efficacy of moxidectin was evaluated in young ponies naturally infected with gastrointestinal parasites. Eight animals were treated orally with moxidectin at 0.4 mg/kg bodyweight and eight received only the vehicle. They were all necropsied two weeks later. Faecal samples were examined daily for egg counts and larval cultures. Parasites were recovered from total faecal samples collected daily and from the gastrointestinal tracts at necropsy. Moxidectin reduced the strongyle egg counts by > 99 per cent from three days after treatment but some individuals remained positive for 10 days. The drug had little or no ovicidal activity. As evaluated in the critical-controlled test, moxidectin was 99 to 100 per cent effective (P < 0.05) against luminal stages of parascaris equorum, Strongylus vulgaris, Triodontophorus species, Craterostomum acuticaudatum, 19 cyathostome species and Oxyuris equi. Adult S edentatus were also completely removed. Its efficacy against third stage larvae of Gasterophilus intestinalis was 95 per cent (P < 0.05). Luminal nematode stages were removed within a few days, and bots continued to be eliminated for at least two weeks after treatment. No activity was observed against Anoplocephala perforliata. As evaluated in the controlled test, moxidectin was 100 per cent effective against Habronema muscae (P < 0.05) and had a 76 per cent but not significant efficacy against encysted small strongyle larvae. PMID- 9854320 TI - Combined outbreak of the genital and conjunctival forms of bovine herpesvirus 1 infection in a UK dairy herd. PMID- 9854321 TI - Contamination of the contents of intact eggs by Salmonella typhimurium DT104. PMID- 9854322 TI - British Veterinary Association. General guidelines on the use of antimicrobials. PMID- 9854323 TI - Free-living wild boar in south-east England. PMID- 9854324 TI - Lungworm infection in dogs. PMID- 9854325 TI - Cutaneous aspergillosis in a babirusa (Babyrousa babyrussa) PMID- 9854326 TI - Key-Gaskell syndrome in cats. PMID- 9854327 TI - Floppy pinnae in Siamese cats. PMID- 9854328 TI - Unilateral epistaxis in two dairy cows. PMID- 9854329 TI - [Effect of remifentanil on clinical and electroencephalographic parameters of depth of anesthesia in balanced anesthesia with propofol, enflurane or isoflurane]. AB - Electrophysiological parameters are well-suited to detect changes in cerebral function. The present study investigates whether balanced anaesthesia with remifentanil during nociceptive stimulation is associated with changes in clinical and electrophysiological parameters indicating inadequate depth of anaesthesia. Following IRB approval and written informed consent, 23 patients (ASA: I; age: 36 +/- 11) scheduled for elective gynaecological laparoscopy were included in the study. Without any premedication, anaesthesia was induced with remifentanil (1.0 microgram/kg bolus injection), propofol (0.5 mg/kg added by repetitive (10 mg) bolus injections every 10 s until unconciousness) and vecuronium (0.1 mg/kg). Following endotracheal intubation (normoventilation: PetCO2: 36 bis 38 mmHg), remifentanil infusion was started with continuous doses of 0.5 microgram/kg/min over 5 minutes and maintained with 0.25 microgram/kg/min during surgery. Remifentanil was randomly combined with propofol (group 1: 100 micrograms/kg/min; n = 7), enflurane (group 2: 0.5 MAC; n = 8) or isoflurane (group 3: 0.5 MAC; n = 8). Monitoring included: heart rate (beats/min), mean arterial pressure (mmHg), oxygen saturation (%), endtidal CO2 (mmHg) and endtidal enflurane and isoflurane (%). EEG: 2-channel recordings of Fz versus mastoid and ECG (artefact control) during steady-state anaesthesia and surgery. Following fast-fourier-transformation (4 s; 256/s; 0.5 to 35.0 Hz), spectral power densities were calculated for the selected frequency bands. Auditory evoked potentials (AEP; middle latency) were registered simultaneously after binaural stimulation via head-phones click-stimulation (6 Hz; 75 dB above hearing threshold; 512 stimulations per average). Bandpass was 0.01 to 2.0 kHz. ANALYSIS: Na, Pa, Nb (latencies; ms) and peak-to-peak amplitudes (NaPa, PaNb; microV). EEG and AEP recording technique [15]. The study protocol included baseline values from pre-intubation, pre-surgery, the respective post-stimulation values (1 min, 3 min, 5 min) and all data at five-minute intervals during surgery until emergence from anaesthesia. During steady-state study conditions with defined remifentanil applications, mean data indicate that in response to nociceptive stimuli no changes in clinical or electrophysiological parameters were observed. In contrast to other studies using different anaesthetic techniques, the present data from remifentanil indicate very stable haemodynamic and electrophysiological parameters (EEG, AEP) during noxious stimulations. Adjustable and with no plasma accumulation, remifentanil demonstrates potent antinociceptive effects resulting in signs of adequate anaesthesia. PMID- 9854330 TI - [New methods for transurethral electroresection of the prostate from the anesthesiologic viewpoint]. AB - Intravascular absorption of large volumes of solution without electrolytes is a common problem in transurethral resection of the prostate. In the present study we compared two different methods of resection (TURP vs. Vaporization TURP; 20 patients in each group) regarding fluid absorption and loss of blood. In addition, we report on 60 patients who had Vapo-TURP. The operations were performed by different very experienced surgeons. In both groups, spinal anaesthesia and standard regimes of infusion were used. The amount of fluid absorbed was measured by the ethanol method (2 vol.% ethanol; Widmark-formula). Haemoglobin, haematocrit and electrolytes were determined at set times. Ten of the 20 patients in the TURP group showed blood-alcohol levels > 0.1%/1000. In the Vapo-TURP group, only five of 57 patients (three patients with perforation of the prostate capsula were excluded) showed positive levels of blood-alcohol (< 0.05%/1000; fluid absorption < 150 ml). The difference was statistically significant (p = 0.002). Sixty minutes after the beginning of the procedures, the values of haemoglobin and haematocrit were significantly lower in the TURP group (7.68 +/- 0.41 vs. 7.38 +/- 0.64 mmol/l and 0.36 +/- 0.02 vs. 0.34 +/- 0.03). Regarding absorption of fluid and bleeding, Vapo-TURP was superior. From the anaesthesiological point of view, Vapo-TURP should be the method of choice, especially for the elderly. PMID- 9854331 TI - [Special aspects of anesthesia in patients with epidermolysis bullosa based on a case example]. AB - Epidermolysis bullosa hereditaria dystrophica (Hallopeau-Siemens) is a rare autosomal recessive disease characterized by extreme bullae formation of skin and mucosa. Typical dystrophic nails and flexion contractures of the joints can lead to deformities. Carious teeth and microstomia caused by scarred contractures of the lips are characteristic of the clinical picture. Depending on the form and severity of epidermolysis bullosa, the anaesthetic and surgical management requires careful planning to avoid unnecessary complications as a result of positioning, anaesthesia or surgery. In cooperation with the patient, optimal positioning on the operating table without pressure or tangential friction of the skin needs to be achieved. Wherever possible, surgical tape and adhesive electrodes should be avoided. Artificial respiration, intubation and monitoring must be adapted to the skin conditions of the patient. In particular the skin below the blood pressure cuff must be protected by adequate padding, and maximum intervals between measurements should be chosen. Nasal, oral, laryngeal and tracheal manipulations should be kept to a minimum for protection of the upper airway. Fibre optic intubation is to be preferred because of the possibility of microstomia and the need for simultaneous airway diagnosis. In addition, oro- or nasopharyngeal tubes and catheters should be avoided where possible. Depending on the course of surgery and anaesthesia, postoperative therapy in an intensive care unit should be considered. PMID- 9854333 TI - Shaping with 'showcase'. PMID- 9854332 TI - [Value of high dosage fluconazole in therapy of candida infections in intensive care medicine]. AB - The administration of fluconazole in the ICU setting in dosages of > or = 800 mg/day or > or = 10 mg/kg/day has been reported in about 400 patients with candidiasis of different localisation including candidemia, with a rapidly increasing incidence of serious candidal infections. In Germany, fluconazole is approved for therapy of life-threatening infections caused by Candida spp. and Cryptococcus neoformans in a dosage of up to 800 mg/day. Especially in non neutropenic patients with life-threatening infections caused by Candida spp., Cryptococcus neoformans and Coccidioides immitis, the results of a limited number of dose-finding trials show dose-dependent response rates. These findings strongly advocate the application of high-dose fluconazole; their evaluation, however, still awaits final clarification. The good safety profile even for maximum dosages of up to 2000 mg/day and the linear, predictable pharmacokinetics up to 1600 mg/day indicate the excellent tolerability of fluconazole in the clinical situation, which justifies prospective, randomized clinical trials with treatment groups as homogeneous as possible for further evaluation of the optimum dosage and duration of treatment in the various types of candidal infection. PMID- 9854334 TI - Oral cancer biopsy in general practice. PMID- 9854336 TI - Standards and dental suppliers. PMID- 9854335 TI - Lignocaine and anaesthetic allergy. PMID- 9854337 TI - Implications of amalgam in general health. PMID- 9854338 TI - Caries prevention--based on evidence? Or an act of faith? AB - The era of evidence-based healthcare requires that dentists' actions are supported by scientific proof of effectiveness. In this paper, evidence regarding the impact of strategies for caries prevention is derived from epidemiology and from a systematic review of research papers. PMID- 9854339 TI - The tarnished history of a posterior restoration. AB - Galvanic corrosion is an electrochemical reaction between dissimilar metals that has the potential to cause unpleasant and even painful biological effects intra orally. A case is presented where a full gold crown underwent galvanic change after being placed in contact with an amalgam restoration. PMID- 9854340 TI - Diabetic hypoglycaemia and the dental patient. AB - Hypoglycaemic attacks in patients with insulin-treated diabetes are unpleasant and inconvenient at best, but may constitute a medical emergency at their most severe. This article provides the dental practitioner with an insight into the physiological processes which occur during such attacks, with clues to the recognition of the signs of impending hypoglycaemia. Steps which may be taken to avoid hypoglycaemia with the various forms of treatment are then considered against a background of their suitability for use in dental practice. PMID- 9854341 TI - Introductory notes regarding a European Medical Risk Related History questionnaire (EMMRH) designed for use in dental practice. AB - The Medical Risk Related History (MRRH) has been in use for several years in the Netherlands. Since 1994 the MRRH system has been subject to research in nine European countries. Legal and ethical demands in all participating countries have been listed, and a national epidemiological analysis of pathology interfering with dental treatment has been undertaken for every participant. PMID- 9854342 TI - Distraction osteogenesis--a review. AB - This review describes the technique of distraction osteogenesis, outlining its application and current developments for the correction of craniofacial anomalies. Also described are some of the difficulties and complications which can arise with speculation on how this may be overcome. Finally, there is an exploration of how this technique might be routinely used to deal with orthognathic, orthodontic and alveolar problems. PMID- 9854343 TI - Colour stability of provisional crown and bridge restoration materials. AB - OBJECTIVE: In an in vitro study to determine the colour stability of provisional restoration materials, the PMMA synthetics Trimm and Cronsin and the bis-acrylic composites Protemp Garant, Protemp Garant NF, Protemp II and Provipont DC were investigated. MATERIALS AND METHODS: The colour changes of the cylindrical untreated samples were measured in comparison with samples after 24 and 72 hours of artificial ageing in a Suntest CPS+ UV ageing device using the Minolta CM 3500d spectrophotometer according to the CIE-L*a*b* system. RESULTS: Trimm (light), Cronsin (brown, yellow and universal), Provipont DC (yellow) and Protemp Garant NF (extra light) displayed the greatest discoloration with values > or = 4 21 delta E, where discoloration toward yellow could be discerned. The remaining materials showed lower luminance reflectance values of < or = 1-4 delta E, and therefore greater colour stability. CONCLUSIONS: Because of their colour stability the materials Cronsin (grey), Protemp Garant (yellow), Protemp II (light), Protemp Garant NF (yellow, light and extra light) could also be used as long-term interim prosthetics. PMID- 9854344 TI - Preventing failed appointments in general dental practice: a comparison of reminder methods. AB - OBJECTIVE: To test the effectiveness of different systems of reminding patients about their appointments in order to reduce the rate of failed attendance. The expense in implementing a reminder system for patients was also estimated. DESIGN: A clinical study in a single-handed dental practice. SUBJECTS: Patients were reminded about their appointment using either postal, manual telephone or automated telephone reminders (or all three combined). A control group received no reminders. 500 patient attendances were recorded in each group. RESULTS: Patients failed to attend for 130 of the 2500 appointments considered in the study. There was a significant reduction in the failed attendance rate from 9.4% (with no reminder) to a minimum of 3% when a reminder was given to the patient before the appointment. However, there was no significant difference among the four reminder test groups, indicating that the form of the reminder made no difference to the failure rate. CONCLUSIONS: Reminding patients using postal or manual telephone techniques is effective at improving attendance. All of the reminder methods, telephone and postal, provided net cost savings rather than additional costs. PMID- 9854346 TI - [Intermittent claudication and nail mycosis. Circulatory disorder as a predisposing illness]. PMID- 9854345 TI - [Diagnosis and therapy of onychomycoses]. AB - Given the four different types of onychomycosis, a large number of differential diagnoses have to be considered. It is therefore imperative that prior to treatment a careful diagnostic evaluation be taken. The simplest methods are the microscopic native (unstained) slide and culture, while more complicated methods are histology, immunofluorescence histology and flow cytometry. For local treatment, antimycotic nail varnishes and urea-containing antimycotic ointments are available, as well as itraconazole, terbinafine and, in the near future, fluconazole, for internal administration. PMID- 9854347 TI - [Irritable bowel: guidelines as an aid for diagnosis and therapy. A continuing education service of the Gastro-Liga]. PMID- 9854350 TI - [Rheumatoid arthritis. 1: Diagnostic criteria and clinical symptoms]. PMID- 9854351 TI - [How is sympathetic nervous system activity evaluated?]. PMID- 9854352 TI - The continuing epidemic. PMID- 9854353 TI - Formocresol in primary teeth. PMID- 9854354 TI - Bilingual journal, bilingual ads? PMID- 9854355 TI - Suggested topics for the Journal and Communique. PMID- 9854357 TI - The Canadian dentists' insurance program: keeping pace with the changing times. PMID- 9854356 TI - The truth about fee guides. PMID- 9854358 TI - Clinical criteria for the use of a decision-making framework for the medically compromised patient: hypertension and diabetes mellitus. AB - In this article, clinical criteria for the staging of disease severity in patients with hypertension and diabetes mellitus are presented. This paper is intended to supplement a previous article by the authors on the use of clinical criteria for the classification of patients with ischemic heart disease and chronic obstructive pulmonary disease and the use of a decision-making framework for the medically compromised patient. Hypertension and diabetes mellitus are discussed in terms of pathophysiology, risk factors, clinical manifestations of disease and disease progression. The article will allow practitioners to stage patients with hypertension and diabetes mellitus and to apply this staging to the previously established clinical decision-making framework for medically compromised patients. PMID- 9854359 TI - The buccal bifurcation cyst: in non-surgical treatment an option? AB - The mandibular buccal bifurcation cyst (BBC) was first described by Stoneman and Worth in 1983. It is a lesion with a specific location, classically the buccal surface of the mandibular first molar and less frequently the mandibular second molar. There have been numerous reports in the literature describing the BBC and its treatment. Treatment advocated thus far includes extraction of the involved first molar, marsupialization and enucleation of the cyst. In their most recent article, Pompura, Sandor and Stoneman reported on the successful treatment of 44 cysts with enucleation without tooth extraction. This article will describe the diagnostic features of the BBC and present three cases with a total of five cysts, which were treated non-surgically and ultimately resolved. The authors, therefore, propose that a more conservative non-surgical approach to these lesions may be considered. PMID- 9854360 TI - [Inter-dentist variability in the provision of fissure sealants]. AB - It has been recognized that the diagnosis and treatment of caries vary so widely among dentists that patients increasingly feel the need to inform themselves and to compare recommended treatments before accepting one. This phenomenon is all the more present in pit and fissure surfaces intervention choices, where there is a wide variety of available treatments. This article will attempt to highlight the factors explaining the pit and fissure treatment variation. The lack of knowledge on pit and fissure sealant effectiveness and indications as well as dentists' clinical experiences have given rise to different myths and prejudices that have had an impact on the variability of treatment decision making. When in doubt about the necessity to execute a treatment, each decision to withhold treatment results in an incomplete care episode. A dentist's lack of comfort with withholding treatment may stop him from offering preventive care and cause him to follow a restoration-oriented practice. It is therefore increasingly urgent to achieve agreement among dentists, especially as current knowledge on the subject is giving way to new research. PMID- 9854361 TI - Quality assurance in the dental profession. AB - The dental profession has traditionally attempted to ensure quality and to guarantee high educational standards by restricting licensure. Recently, many factors have increased public and government interest in the quality of care provided by health professionals. A framework for understanding quality assurance is presented. This framework incorporates with Donabedian's dimensions of structure, process and outcome the additional dimensions of system capacity and community. All dimensions of quality are assessed by comparison with an accepted standard. Quality assurance can be considered as occurring at a system level, a managerial level, and a practice level. All quality assurance activities work to fulfil society's requirements to maintain self-governing professional status. PMID- 9854362 TI - Dental composite biomaterials. PMID- 9854364 TI - Towards optimized treatment outcomes for dental implants: a conference report. PMID- 9854363 TI - Why do we need education in geriatric dentistry? AB - The elderly are the fastest growing segment of the population, both in number and as a percentage, throughout the world. The challenges of dealing with the increased need for oral health care in older adults requires a holistic approach in order to deal not only with there clinical needs, but with there social, functional and communication needs as well. Undergraduate and graduate programs in dentistry in Canada should focus on all the needs of the geriatric population. This is essential if future dental professionals are to gain an understanding of the true dental health care requirements of the elderly and acquire the knowledge, experience and willingness to provide appropriate care for their varied and growing geriatric clientele. PMID- 9854365 TI - [Assisted fertilization in male sterility is not without risks. Genetic aberrations can be transmitted to children and grandchildren]. PMID- 9854366 TI - [Transplantation dilemma: the effort to find organs requires ethical consideration]. PMID- 9854367 TI - [Why don't we protest?]. PMID- 9854368 TI - [Evolution as a philosophical argument]. PMID- 9854369 TI - [Is patient's profession without significance?]. PMID- 9854370 TI - [IT in health care threatens confidentiality and integrity. The Little Brother of the network can see everything about you]. PMID- 9854371 TI - [New knowledge of the mysterious "frozen shoulder". Surgical treatment can accelerate the recovery in more serious cases]. AB - Frozen shoulder (adhesive capsulitis) affects 2-5 per cent of the population, but is most common in the 40-60-year-old age group. The disorder is divided into three phases, the painful, the stiff and the recovery phases. In most cases the condition is self-limiting with negligible residual manifestations, though its average duration is about 30 months. New findings suggest frozen shoulder to be a Dupuytren-like disorder. Pain relief and physiotherapy are usually sufficient, but in more severe cases manipulation with the patient under anaesthesia, possibly combined with distension arthrography or arthroscopic release, may yield rapid improvement in shoulder function. PMID- 9854373 TI - [Male infertility is often of genetic origin. Y chromosome deletions are discovered by the PCR technique]. PMID- 9854372 TI - [Dosage of anesthetics--a tricky question]. AB - The optimal dosage of anaesthetic agents may be difficult. Not only does the intensity of surgical stimuli vary during a surgical procedure, but drug sensitivity varies between subjects exposed to comparable stimuli. Moreover, clinically monitored body reactions do not always reflect the balance between central nervous system effects of the surgical stimuli and of the anaesthetic agent. Therefore, the specialist in anaesthesiology requires access to additional methods of monitoring to enable dosage to be optimised for each patient, minute by minute, thus improving the chances of maintaining an appropriate depth of anaesthesia. Two electro-encephalographic techniques are presented in the article, and aspects of under- and over-dosage of anaesthetic agents are discussed. PMID- 9854374 TI - [Increased birth weights in the Nordic countries. A growing proportion of neonates weigh more than four kilos]. AB - The proportion of children with high birthweights is increasing in Sweden, as in the other Nordic countries. According to the Swedish national birth registry (founded in 1973), the proportion of term (i.e. (37 gestational weeks) offspring of singelton pregnancies, and weighing four kg or more, increased from 16.9 per cent in 1973 to 20.3 per cent in 1995. The respective figures for the first-born subgroup were 12.9 and 15 per cent, the increase in mean birthweight being from 3400 to 3520 g. There was a corresponding increase in head circumference. The risk of delivery-related complications increases with birthweight over four kg, and a higher incidence of major perineal rupture (grade 3 or 4) has been reported, as well as a disturbing increase in the incidence of brachial plexus damage. Findings in recent studies suggest high birthweight to be associated with an increased risk of subsequent morbidity, both in childhood and in adulthood, specifically diabetes type 1, eczema and certain malignancies, particularly breast cancer and prostate cancer. Although the cause of the increasing proportion of large newborns is not known, it may be partly due to weight increase among gravidae. Another possible explanation is reduced maternal smoking, as smoking is less common now than in the 1970s. PMID- 9854375 TI - [Genetic origin of malignant hematopoiesis well defined]. AB - It is vital to determine which cell lines are affected in haematological malignancies, since such information is important to an understanding of the biology of neoplastic stem cells and their capacity to differentiate and mature. Parallel studies of cellular morphology and of chromosomal anomalies is an approach permitting determination of lineage specificity for different haematological neoplasms. Findings in current studies suggest that acute myeloid leukaemia and myelodysplastic disorders generally involve cells of myeloid lineage only, whereas myeloproliferative disorders may also involve lymphoid cell lines. Lymphoid malignancies such as non-Hodgkin's disease or acute lymphoid leukaemia usually involve lymphoid cell lines. PMID- 9854376 TI - [Guidelines for drug information on the Internet]. PMID- 9854377 TI - [A review of the current research on prions. The evidence suggests the possibility of transmission of the mad cow disease to humans]. AB - Further evidence of the transmissibility of bovine spongiform encephalopathy (BSE) across the species barrier from cow to man has been derived from epidemiological analysis and the characterisation of prion strains. Recent research has shown the persistence of prions after experimental transmission to resistant murine species, and subclinical persistence in cows. The accumulation of pathological prion proteins in tonsils and appendix has been demonstrated prior to clinical confirmation of the presence of the new variant of Creutzfeldt Jakob disease. Current prion research is focused on the involvement of B lymphocytes as carriers, on the species barrier and cellular receptors, and on macromolecules involved in the conformational change from normal to pathological prion proteins. PMID- 9854378 TI - [Despite better knowledge of pathogenesis and treatment: atopic eczema--a disease with increasing incidence in Sweden]. AB - The past twenty years have witnessed an increasing incidence of atopic dermatitis in Western Europe. The article consists in a discussion of the pathogenesis, clinical signs and treatment of this common skin disease. Both an IgE-mediated reaction on epidermal Langerhans cells, and a physiological/biochemical defect of the skin barrier structure may be crucial factors of the multifactorial pathogenesis. Local treatment with corticosteroids and moisturisers remains the basic approach, though the development of new more specific treatments is under way. Although much remains to be learned about atopic dermatitis, today all patients can be offered effective treatment resulting in improved quality of life. PMID- 9854379 TI - [The theriaca--a potent antidote and a universal agent]. PMID- 9854380 TI - [Ethics in sports medicine--health and/or performance?]. AB - Ethics in the field of sports medicine embraces a number of issues of general human interest. While it is important to determine the salubrious potential of exercise and sports, it is open to question whether it is reasonable to invest massive research resources in attempts to induce high-performance elite athletes to perform yet better, or to remedy sports injuries to enable athletes to continue exposing themselves to the risk of further injuries. When such questions arise as the extent to which physicians should attempt to modify unhealthy lifestyle factors, for example the use of doping, or whether it is right that athletes should take precedence in waiting lists for health care, it is important to be aware of the singular practical potential of sports medicine. PMID- 9854381 TI - [The SBU on smoking cessation. A simple question and five minutes' advice is an effective method]. PMID- 9854382 TI - Dance to the death. PMID- 9854383 TI - What's wrong with the British Judicial System? PMID- 9854384 TI - Deaths in police custody. PMID- 9854385 TI - Disease, drugs and alcohol induced driving impairment: the law and the medicine. PMID- 9854386 TI - [Use of personal medical data for controlling health care costs: does the end justify the means?]. PMID- 9854387 TI - [Advanced glycation end products or Maillard products]. PMID- 9854389 TI - [Epidemiology and risk factors of Alzheimer's disease]. AB - The epidemiology of Alzheimer's disease is complex. The approach of the distribution of this disease in human populations is limited by an insidious onset and a difficult diagnosis assessment for alive patients. The hunt for Alzheimer's disease determinants has been boosted by the development of molecular biology techniques, allowing the detection of genetic susceptibility factors. Epidemiology offers new areas of research in the field of a disease whose impact and severity is growing, but whose preventive and therapeutic issues remains uncertain. PMID- 9854388 TI - [Brain lesions, pathogenic and etiologic hypotheses of Alzheimer's disease]. AB - The main lesions of Alzheimer's disease are: 1. amyloid deposits, labelled by antibodies directed against the A beta peptide (core of the senile plaques, diffuse deposits and amyloid angiopathy), 2. neurofibrillary lesions labelled by anti-tau antibodies (neurofibrillary tangles, neuropil threads, crown of the senile plaques) and 3. loss of neurons and synapses. The distribution of neurofibrillary pathology is hierarchical: they begin in the entorhinal cortex, progress along the anterograde corticocortical pathways toward the multimodal and unimodal associative cortices to reach, in the most severe cases, the primary cortices. Amyloid lesions are more diffuse, rapidly affecting all the cortical areas. The density of neurofibrillary tangles in the cerebral cortex is correlated with the severity of dementia. Neuritic plaques, synaptic and neuronal loss also contribute to the intellectual deterioration. There are various causes of Alzheimer's disease (several mutations, trisomy 21, repeated head trauma as in dementia pugilistica): it should be considered a syndrome. Its pathophysiology is complex and involves several proteins (e.g. amyloid protein precursor, tau protein, presenilins 1 and 2, and apolipoprotein E). PMID- 9854390 TI - [Early diagnosis of Alzheimer's disease]. AB - The diagnosis of dementia of Alzheimer type (DTA) in the early stage of the disease may be possible with the development of specialized memory consultations. After this diagnosis, we can inform family and even the patient in some cases, monitor the progression of disease, give or not treatment, and help family for daily life management (money, car driving, administration papers, etc.). The first step is to analyse the memory complaints of the patient and if possible of his family. The difficulties in everyday life are more informative than an anxious complaint of memory and some IADL deficiency appear early: difficulty to use phone, drugs, transports or checks. The patient examination by a practitioner or a neurologist has to be complete, even if normal in most of the cases. The practitioner has to explore without specific material, the main cognitive domain: the "5 words test" illustrates a useful memory tool, analysing two types of recall. We also must observe writing, digit span, naming and verbal fluency. The examination by a psychologist using standardized batteries of tests must always be driven by this clinical exploration using common scales like MMS or ADAS. In very early DTA, we may observe a cognitive decline limited to some aspects of memory (free long term recall, cued recall, working memory...) with a decline of the most controlled attentional tasks. An isolated memory disorder has to be identified either as onset of DTA or as an aging related cognitive decline (ARCD): the difference is qualitative and may appear only on repeated exams by experimented psychologist. PMID- 9854391 TI - [Cognitive manifestations of Alzheimer's disease]. AB - Cognitive features in dementia of the Alzheimer's type (DAT) appear highly heterogeneous. Neuropsychological models seem useful to characterize the cognitive decline. Functional imaging may identify anatomoclinical correlations. Memory impairment is the major symptom in DAT, associated with cortical dysfunctions (aphasia, apraxia, agnosia) and deficits in executive functions. Various evolutive profiles increase this clinical heterogeneity. The use of standardized and validated psychometrical tools appears very important for diagnosis and follow-up of cognitive decline. The MMSE, for example, deserves to be included in the everyday clinical practice. PMID- 9854392 TI - [Mental-behavioral disorders in Alzheimer's disease]. AB - Behavioural and psychological disorders are often observed in Alzheimer's disease. Some are common, such as symptoms of depression, apathy, aggressivity, agitation, psychotic disturbances, disorders of sleep rhythm, etc. Many factors contribute to the aetiology of these disorders, mainly cerebral lesions, environmental changes, somatic illnesses, iatrogenic factors and psychological reaction mechanisms. Management must take each into account. Treatment comprises medication (symptomatic treatment of the various disorders, cholinergic treatment) and other means (adaptation of the inhabitation, informing family and friends, psychotherapy, etc.). PMID- 9854393 TI - [Differential diagnosis of Alzheimer's disease]. AB - The differential diagnosis of Alzheimer's disease has dramatically changed since the evolution of the diagnostic strategies (with definition of neuropsychological, behavioural, and imaging patterns) and disposal of cholinergic drugs indicated in Alzheimer's disease. The question is no more centred on exclusion of the traditional reversible dementias or depression. It is centred on the distinction between Alzheimer's disease and other degenerative diseases still often misdiagnosed with Alzheimer's disease such as frontotemporal dementias, dementia with Lewy bodies, and some focal atrophies that do not have the same physiopathology and should not be treated with anticholinesterase drugs. Besides, better knowledge on the links between cerebrovascular pathology and Alzheimer's disease, remind us that both pathologies may coexist and should be taken into account. PMID- 9854395 TI - [Non-drug management of Alzheimer's disease]. AB - The recent appearance of drugs offering some efficacy has been a major advance in the treatment of Alzheimer's disease. However, non drug therapy of the patients and caring for family members remain the main elements to delay institutionalization. General practitioner has a major role in this approach whose aims are identification and treatment of excess disability, understanding and control of psycho-behavioral manifestations, sustaining and stimulation of cognitive functions and communication, maintaining the patient's autonomy and alleviating of the caregiver's burden. A major point of this approach is to consider the disease manifestations not only as the consequence of brain lesions but also as the impact of the disease on the affective and relational functions of the subject. PMID- 9854394 TI - [Drug therapy strategies in Alzheimer's disease]. AB - Treatments in Alzheimer's disease include treatment of cognitive impairment and behavioral manifestations (agitation, depression, anxiety, delusions). It should be noted that many non cognitive behaviors may have some relations to underlying cognitive impairment. In the not too distant future, physicians can expect to see a variety of medications and controversies over the benefits of slowing symptoms with cholinergic therapeutics approved for clinical use and (or) preventing progression of Alzheimer's disease assessed in clinical trials will emerge. PMID- 9854396 TI - [Alzheimer's disease. Clinical practice]. PMID- 9854397 TI - [4 figures and a commentary]. PMID- 9854398 TI - [Oligoarthritis or polyarthritis of less than 3-month duration. Diagnostic trends]. PMID- 9854400 TI - [Closed thoracic injuries: physiopathology, diagnosis, management in emergencies]. PMID- 9854401 TI - [Cauda equina syndrome]. PMID- 9854399 TI - [Cerebrovascular stroke: epidemiology, etiology, physiopathology, diagnosis, course, treatment]. PMID- 9854402 TI - [Acute ischemia of the lower limbs: etiology, diagnosis and management in emergencies]. PMID- 9854403 TI - [Echocardiography: indications]. AB - Echocardiography is the most important non-invasive test for many cardiovascular diseases. Morphologic and haemodynamic evaluation can be performed in a short time. In this review we will discuss the most common and the most important indications for an echocardiographic assessment. A detailed list for a quick reference is included at the end. PMID- 9854404 TI - [Secondary prevention of ischemic heart disease after coronary revascularization]. AB - This study was aimed to assess the compliance with policies for secondary prevention of coronary heart disease (CHD) one year after coronary artery revascularization with special attention to the management of hyperlipidemia. One year after coronary revascularization during the year 1994, patients were contacted by letter to determine the modification of their risk factors, the treatment patterns for hypercholesterolemia and to have their plasma lipid level and blood pressure measured. Of the 245 consecutive patients contacted (110 after coronary artery bypass grafting, and 135 after percutaneous transluminal coronary angioplasty), 186 (76%) provided the information required for further analysis. Excluding the patients older than 65 years, only 29 out of 97 patients (30%) with a total cholesterol of more than 5.2 mmol/l, and only 20 out of 52 patients (38%) with a total cholesterol of more than 6.2 mmol/l were receiving lipid lowering therapy 1 year after coronary artery revascularization. In contrast, 97% (n = 180) of the entire population studied were taking antiplatelet drugs and/or coumadine. Participation in an in-house rehabilitation program yielded a positive influence on smoking, but not on treatment of hypercholesterolemia. In conclusion, only a small proportion of patients with documented CHD and hypercholesterolemia were being treated for their lipid disorder 1 year after coronary artery revascularization. In contrast, the great majority of patients received antiplatelet and/or coumadine therapy: These results indicate that the compliance with published treatment guidelines for hyperlipidemia in patients with CHD is still highly inadequate, irrespective of the participation in a rehabilitation program. PMID- 9854405 TI - [Clinical picture of acute right heart infarct]. PMID- 9854406 TI - [Acute rupture of a solitary angiomyolipoma in the upper pol of the right kidney with extensive perirenal hematoma]. PMID- 9854407 TI - [Exertional dyspnea in HIV infection]. PMID- 9854408 TI - [Arterial hypertension with concomitant hypokalemia]. PMID- 9854409 TI - Eliminate the J-1 visa waiver program. PMID- 9854410 TI - Being right is being ethical. PMID- 9854411 TI - Questioning the success. PMID- 9854412 TI - Worry about patients, not profits. PMID- 9854413 TI - Putting patients first. PMID- 9854414 TI - MedBytes. PMID- 9854416 TI - Medical privacy? PMID- 9854415 TI - Forum on ethics. Big Brother versus Hippocrates: no more privacy? PMID- 9854417 TI - Setting the record straight. PMID- 9854418 TI - Surviving a Titanic ride. PMID- 9854419 TI - Texas breaks new ground. PMID- 9854420 TI - How do you stack up? PMID- 9854421 TI - Tick-borne diseases in Texas: a 10-year retrospective examination of cases. AB - The occurrence of Rocky Mountain spotted fever, human monocytotropic ehrlichiosis, tularemia, tick-borne relapsing fever, and a southern erythema migrans-like illness (Lyme/Lyme-like disease) is determined by the geographic distribution and seasonal activity of the particular vector tick(s). The flulike signs and symptoms early in the course of spotted fever rickettsiosis, ehrlichiosis, tularemia, and relapsing fever are nonspecific and do not readily suggest a particular diagnosis. Laboratory diagnosis, particularly during the acute stage of illness, often is elusive. Empiric treatment with doxycycline can be lifesaving for Rocky Mountain spotted fever and ehrlichiosis. This article provides insight into the significance of each of these tick-transmitted diseases in Texas. PMID- 9854422 TI - Macroamylasemia in HIV infection. AB - A case of macroamylasemia was seen in a 40-year-old HIV-positive bisexual male treated at the Fort Worth-Tarrant County Health Department (Ryan White Clinic). Macroamylasemia is a rare condition encountered sometimes in persons with HIV infection. Apart from the setting of HIV infection and acquired immunodeficiency syndrome, macroamylasemia is seen also in various conditions including liver disease, diabetes, cancer, malabsorption, and autoimmune disorders. Although this biochemical phenomenon requires no therapy, it should be considered in the differential diagnosis of patients who have persistently high levels of serum amylase and yet do not exhibit any clinical symptoms of pancreatitis or salivary gland inflammation. PMID- 9854423 TI - Synthetic peptides: a modern approach to vaccination. AB - For induction of humoral or cell-mediated immunity, development of synthetic peptide vaccines present a novel approach and a better alternative to conventional vaccines. Construction of synthetic peptides that mimic antigenic sites is the basis of this approach. In this article various methodologies involved in the peptide synthesis are discussed. This is followed by a discussion on the strategies involved in rendering poor or non-immunogenic peptides immunogenic. These include coupling to large carrier proteins, polymeric presentation and incorporation of an identified Th cell epitope into the peptide. PMID- 9854424 TI - Immune response of electroeluted detergent soluble 29 kDa antigen from Setaria digitata (Von Linstow). AB - Filariasis is one of the typical parasitic infections which cause immune suppression during the course of infection in both humans and experimental animals. A 29 kDa protein isolated from detergent soluble antigen of S. digitata showed maximum inhibition of cell mediated immune response. The heat inactivated 29 kDa protein was found to be devoid of property of suppression of immune response in the host. Histological study of spleen of BALB/C mice immunized with 29 kDa protein showed changes in regions of spleen such as follicle, trabeculae, capsule, reticuloendothelial cells and eosinophils. The 29 kDa protein, the most reactive of the detergent soluble proteins produced partial suppression of immune response, thereby contributing to the factors responsible for the survival of filarial parasites in hosts. PMID- 9854425 TI - Bacterial infection and endotoxin in female reproductive tract in rats: correlation with the developmental status of preimplantation embryos. AB - During mammalian preimplantation development, a substantial numbers of embryos are believed to be lost for reasons that are unclear. Using female rats, we investigated whether the developmental status of embryos is influenced by bacterial infection and endotoxin in the reproductive tract. From the vagina of cycling rats (n = 11), 21 bacterial isolates were identified; they were Streptococcus faecalis (S. faecalis; 38%), Escherichia coli (E. coli; 19%), Acinetobactor calcoaceticus (A. calcoaceticus; 14%), and coagulase negative staphylococcus (14%), Micrococcus sp. (5%), Bacillus subtilis (B. subtilis; 5%) and Proteus vulgaris (P. vulgaris; 5%). From the vagina of day 4 pregnant rats (n = 12), 26 isolates were identified; they were S. faecalis (23%), A. calcoaceticus (23%), E. coli (15%), Micrococcus sp. (15%), B. subtilis (8%), P. vulgaris (4%), Staphylococcus aureus (4%), beta-hemolytic streptococcus (4%) and Pseudomonas aeruginosa (4%). Gram negative bacteria found in the vagina of cycling and day 4 pregnant rats were 38% and 46%, respectively. In both, bacterial load was 10(3) 10(5) colony forming units and there was no association with the abnormality of the recovered embryos. However, in two day 4 pregnant animals, pathogenic bacteria (Staphylococcus aureus and beta-hemolytic streptococcus) were isolated and embryos recovered from them were degenerated and deformed. The vagina of day 9 pregnant animals (n = 7) were, however, sterile. Consistently, in all animals, the upper reproductive tract (uterus and oviduct) was devoid of any bacteria and no anaerobic bacteria were isolated from any part of the tract. The levels of endotoxin in the vagina of cycling and day 4 pregnant rats were 1.35 +/- 0.1 and 1.17 +/- 0.1 endotoxin units (EU), respectively. It was undetectable in the oviduct and uterus of all animals (n = 5) except one which showed high levels of endotoxin in uterus (4.5 EU) and oviduct (2.2 EU) and the animal also produced degenerated and deformed embryos. These results indicate that common bacterial flora of vagina may not affect embryo development and the presence of pathogenic bacteria in the vagina and/or endotoxin in reproductive tract could be detrimental to viability of gametes and preimplantation embryos in rats. PMID- 9854426 TI - Immunoexpression of mutant p53 and human papillomavirus related E6 oncoprotein in anal malignancies. AB - Immunohistochemical expression of mutant p53 protein and human papillomavirus (HPV) 16 and 18 related E6 oncoprotein was studied in 36 biopsy proved anal cancers. Mutant p53 was detected in 61.1% cases. HPV 16 and 18 E6 protein was expressed in 22.2% cases, all of which were squamous cell carcinomas. Coexpression of both mutant p53 and E6 protein was found in only 5 cases (13.8%). In HPV 16/18 positive anal tumors, the degradation of p53 is accelerated by viral E6 oncoprotein. In HPV negative tumors, however, other mutagenic factors probably play a role in carcinogenesis. PMID- 9854427 TI - Nucleotide sequence of DR-B exon of major histocompatibility complex from an Indian zebu cattle breed. AB - The Indian cattle is known to be more tolerant to tropical infections than the European cattle. In order to verify the genetic basis of this variation, the DR B exon-2 of the major histocompatibility locus, known for coding the antigen recognition site, from the Hallikar breed of Indian cattle was amplified by PCR, cloned and sequenced. Comparison of this sequence with the information available on taurus cattle brought out six unique nucleotide changes and three amino acid changes. The amino acid positions were at 17, 72 and 87. A major variable region was observed at amino acid position 85 to 87 from all the alleles so far reported for the bovine locus. PMID- 9854428 TI - 5/6/7/8 mono or disubstituted 1H/1-phenyl-9H-pyrido [3,4 b] indoles--a new class of antifilarial agents and their relationship with voltage sensitive calcium channels in Setaria cervi. AB - Synthetic compounds of 5/6/7/8 mono or disubstituted 1H/1-phenyl-9H pyrido (3,4 b) indole series were screened for their antifilarial activity in vitro by observing their effect on the spontaneous movements of the whole worm and nerve muscle preparation of S. cervi (Nematoda:Filarioidea). Of the 10 compounds tested only compound 87/581 was able to modify the movements of whole worm and nerve muscle preparation. This compound caused inhibition of spontaneous motility of whole worm and nerve muscle preparation of S. cervi characterized by initial short lasting stimulation followed by irreversible paralysis. The concentration required to inhibit the movements of n.m. preparation was 10 micrograms/ml as compared to 25 micrograms/ml for the whole worm preparation, suggesting a cuticular permeability barrier. Prior addition of compound 87/581 to the bath fluid blocked the calcium ion dependent stimulant effect of Ach and pyrantel pamoate. Further the initial stimulant effect of the compound was blocked by prior addition of nifedipine a calcium channel blocker. The findings suggest that the action is due to blockade of voltage sensitive calcium channels. PMID- 9854429 TI - Effect of plumbagin on the radiation induced cytogenetic and cell cycle changes in mouse Ehrlich ascites carcinoma in vivo. AB - The effect of plumbagin, a naphthoquinone from the roots of the Indian medicinal plant Plumbago rosea, and Cobalt-60 gamma radiation was studied on Ehrlich ascites carcinoma in vivo, taking cytogenetic damage and cell cycle changes as experimental endpoints. Plumbagin (5 mg/kg body wt, P1) administered intraperitoneally produced a significant increase in the percentage of S-phase as well as G2-M cells with a corresponding decrease in the G1 phase at different post-treatment times. Radiation (7.5 Gy, RT) alone produced the classical G2 block at 1 hr, which persisted with a continuous increase throughout the post treatment observation period. The combination treatment produced a similar effect as that of RT on G2-M cells, but its effect on the G1 phase was more pronounced than the latter. While P1 treatment produced a small increase in the percentage of labeled S-phase cells, combination treatment significantly reduced the labeled S-phase cells with a corresponding increase in the unlabeled fraction. Drug or radiation alone significantly increased micronuclei induction at various post treatment times and the combination of the two further enhanced this effect additively. The mechanism of interaction of P1 with radiation in bringing about this effect is not clear. PMID- 9854430 TI - Influence of Aloe vera on collagen turnover in healing of dermal wounds in rats. AB - Treatment of full-thickness wounds with A. vera, on rats resulted in increased biosynthesis of collagen and its degradation. A corresponding increase in the urinary excretion of hydroxyproline was also observed. Elevated levels of lysyl oxidase also indicated increased crosslinking of newly synthesised collagen. The results suggest that A. vera influences the wound healing process by enhancing collagen turnover in the wound tissue. PMID- 9854431 TI - Antioxidant effect of two flavonoids from the bark of Ficus bengalensis Linn in hyperlipidemic rats. AB - Two flavonoid compounds, viz. 5,7-dimethyl ether of leucopelargonidin 3-0-alpha-L rhamnoside and 5,3'-dimethyl ether of leucocyanidin 3-0-alpha-D galactosyl cellobioside obtained from the bark of F. bengalensis were evaluated for their antioxidant action in hyperlipidemic rats. The results were compared with the activity of a structurally similar flavonoid, quercetin, a known antioxidant. The Ficus compounds showed significant antioxidant effects which may be attributed to their polyphenolic nature. The methylation of two hydrozyl groups in the Ficus flavonoids might have slowed down their antioxidant action as compared to quercetin. PMID- 9854432 TI - Inhibitory effect of Garcinia kola on lipid peroxidation in rat liver homogenate. AB - Garcinia kola, (a herb grown in Nigeria; calorific value 358.54 k.cal/100 g) inhibited in vitro lipid peroxidation of rat liver homogenate in a dose dependent manner. The inhibitory activity of G.kola was not affected by heating (100 degrees C/10 min). The antioxidant component of G.kola was soluble in aqueous and ethanolic media. The active component(s) in G. kola responsible for its inhibitory activity on lipid peroxidation is tentatively identified as isoflavones. PMID- 9854433 TI - Effect of bacterial association on virulence of Entamoeba histolytica to baby hamster kidney cell monolayers. AB - Axenic E. histolytica trophozoite strain NIH:200 and HMI:IMSS when co-associated with aerobic bacteria Escherichia coli strain K12 and serotype 056 showed marked increase in virulence as observed by destruction of baby hamster kidney (BHK) monolayers. However, when incubated with anaerobic bacterial strains Clostridium perfringens and Bacteroides fragilis virulence remained unaltered. Further, adherence of E. histolytica to BHK monolayer was found to be mediated by N-acetyl D-galactosamine. PMID- 9854434 TI - Electrophoretic analysis of polypeptides and glycopeptides of erythrocyte membrane sampled from rats simulating mild insulin dependent diabetes mellitus. AB - In order to understand the molecular mechanism of reduced life span of diabetic erythrocyte, polypeptides and glycopeptides were analyzed by disc gel preparative sodium dodecyl sulphate polyacrylamide gel electrophoresis. An additional glycopeptide (244.5 kDa) and two additional polypeptides (39.81 and 144.5 kDa) were observed on glycopeptide and polypeptide gel profiles of mild insulin dependent diabetes mellitus (mIDDM) sample as compared to control. On the basis of molecular weight, their position on gel profile and their widely accepted nomenclature they were termed as glycosylated-ankyrin, membrane accreted glyceraldehyde-3-phosphate dehydrogenease (G 3-PD) and stress induced band 2.3 peptide. Earlier we have reported an increase in heterogeneity associated with increase in the population of aged fragile cells having altered membrane bound cation dependent ATPases, cytosolic dehydrogenase and hexokinase activities of mIDDM simulating rat erythrocyte sample. Significance of above observation in view of our earlier observation is discussed to explain the molecular mechanism of reduced life span of diabetic erythrocytes. PMID- 9854435 TI - Mammalian coronary perfusion--an improved laboratory device. AB - An improvised device for coronary perfusion of isolated rat heart consisted of a reservoir of buffer pressurised with continuous bubbling of O2 connected to two double condensors in series. The outer jacket of each condenser was connected to a temperature controlled motorised water bath to ensure steady temperature of buffer. Isolated hearts could be tested with known agents directly through a syringe connected to the aortic cannula. PMID- 9854436 TI - Assessment of carbonic anhydrase activity in blood by alteration in pH. AB - Carbonic anhydrase enzyme activity could be measured by manometeric and colorimetric techniques. A simpler and modified method of carbonic anhydrase enzyme activity assessment in blood is proposed. In the present method differences in pH by hydration of CO2 in absence and presence of carbonic anhydrase inhibitor have been used to measure the carbonic anhydrase activity in the blood. PMID- 9854437 TI - Reinforcement without awareness: I. Voice level. AB - Although some studies have reported that reinforcement can strengthen people's behaviour without their awareness, most studies that have incorporated adequate procedures for assessing awareness have been unsuccessful. Using rigorous procedures, we report two studies designed to provide more scope for learning without awareness by making the reinforcement contingency as unobtrusive as possible. Subjects were told that the experiment was on ESP, and that they were to say which of two words the experimenter was thinking about. In fact, reinforcement was contingent on the loudness of the subject's voice when responding. Even though subjects reported no awareness of a relationship between trial outcomes and their behaviour, the probability of the reinforced responses increased significantly over trials in both experiments. These results support the view that reinforcement can occur without awareness. PMID- 9854438 TI - Reinforcement without awareness: II. Word class. AB - In a previous study of reinforcement without awareness by Lieberman, Sunnucks, & Kirk (this issue), subjects were told that the experiment was on ESP; two words were presented on every trial, and their task was to choose the word the experimenter was thinking of. In fact, reinforcement was contingent on the loudness of the subjects' voices when responding. They found stronger evidence for reinforcement without awareness than in many previous studies, and they attributed their success in part to the reinforcement of a response to which subjects were unlikely to attend. To explore this factor further, we again used an ESP cover story but reinforced subjects for choosing the word in each pair that contained a double letter. We found evidence of reinforcement without awareness in three experiments and also identified two factors that influence these effects: Learning did not occur if subjects (a) were encouraged to test irrelevant hypotheses, or (b) sat in an uncomfortable chair. We speculate that learning without awareness may be more likely when subjects are relaxed and that hypothesis testing and uncomfortable chairs impair such learning because they prevent relaxation. PMID- 9854439 TI - Simultaneous discrimination learning in pigeons: value of S- affects the relative value of its associated S+. AB - In a simple simultaneous discrimination involving a positive stimulus (S+) and a negative stimulus (S-), it has been hypothesized that positive value can transfer from the S+ to the S- (thus increasing the relative value of the S-) and also that negative value can transfer from the S- to the S+ (thus diminishing the relative value of the S+; Fersen, Wynne, Delius, & Staddon, 1991). Evidence for positive value transfer has been reported in pigeons (e.g. Zentall & Sherburne, 1994). The purpose of the present experiments was to determine, in a simultaneous discrimination, whether the S- diminishes the value of the S+ or the S- is contrasted with the S+ (thus enhancing the value of the S+). In two experiments, we found evidence for contrast, rather than value transfer, attributable to simultaneous discrimination training. Thus, not only does the S+ appear to enhance the value of the S-, but the S- appears to enhance rather than reduce the value of the S+. PMID- 9854440 TI - Determinants of positive and negative generation effects in free recall. AB - Better retention of self-produced as opposed to experimenter-presented material is called generation effect; the reverse phenomenon is the negative generation effect. Both are found in intentional-learning experiments in which generating versus reading is manipulated between subjects. The present article presents an overview of those findings and aims at clarifying the conditions under which these effects emerge. Experiments 1 and 2 demonstrate that if cue-target relations are manipulated within one list, a negative generation effect in free recall can be obtained for all items, no matter which cue-target relation they bear. In Experiment 3, cue-target relations were manipulated between lists. Here, a negative generation effect in free recall was found only in lists in which items were cued with words that mismatched the inter-target relations, whereas a positive generation effect was observed in those lists in which the generation cues matched the inter-target relations. A subsequent cued-recall test demonstrated that in cases of mismatch of relations, participants in the generate condition process cue-target relations at the expense of inter-target relations. The three-factor theory can be integrated with the task-demand account in a transfer-appropriate processing framework to accommodate these findings. PMID- 9854441 TI - Period duration of physical and imaginary movement sequences affects contralateral amplitude modulation. AB - The hypothesis was tested that the strength of cross-manual effects of voluntary amplitude modulations in bimanual tasks increases when less preparation time is available during the execution of a sequence of movements. By means of the continuation procedure, various period durations (600, 800, 1200, and 1800 msec) of movement sequences were imposed. Subjects performed bimanual periodic arm movements on two digitizers: constant-amplitude movements (short or long) with the one hand, and movements of constant short or constant long amplitudes and movements of alternating short and long amplitudes with the other hand. Period duration-dependent modulations of the amplitude were observed in movements of instructed constant amplitudes when the other hand performed alternating amplitude movements, but not when the other hand performed constant-amplitude movements. Not only physical performance, but also the imaginary production of alternating-amplitude movements resulted in period-duration-dependent cross manual effects, though reduced in size. The pattern of results is in agreement with a recently proposed two-level model of cross-manual effects according to which cross-talk can occur at the programming level as well as at the execution level. PMID- 9854442 TI - Random generation and the executive control of working memory. AB - A series of experiments explores the capacity for generating sequences of random responses, relating it to the central executive component of working memory. Experiment 1 shows a broadly similar pattern of redundancy increasing with speed of generation for both the verbal generation of digits and the manual pressing of keys. In both cases deviations from randomness are shown to reflect the increasing use of a limited number of stereotyped response sets. The remaining experiments use keyboard generation. Experiment 2 demonstrates that concurrent immediate serial recall decreases randomness, and that longer recall sequences produce less random output. Experiments 3 and 4 show that whereas simple counting has no effect on randomness, serial recall, semantic category generation, and concurrent digit generation have substantial effects, and a concurrent fluid intelligence test has the greatest influence on the randomness of key pressing. It is suggested that the task of random generation resembles that of category fluency because it requires the subject to switch retrieval plans and inhibit repetition. On this basis it is predicted that a task involving repeated switching of categories will interfere with generation, despite being predictable and having a low memory load. Experiments 5 and 6 confirm this prediction. Strengths and limitations of the switching hypothesis are discussed, as are the implications of our results for the analysis of executive processes. PMID- 9854443 TI - The representation of sublexical orthographic-phonologic correspondences: evidence from phonological dyslexia. AB - Although there is considerable evidence that grapheme and body units are involved in assembling phonology from print, there is little evidence supporting the involvement of syllabic representations. We provide evidence on this point from a phonological dyslexic patient (ML) who, as a result of brain damage, is relatively unable to read nonwords. ML was found to be able to perform tasks assumed to reflect processes involved in assembled phonology (i.e. segmentation, orthographic-phonologic conversion, and blending) when the units involved were syllables, but demonstrated considerable difficulty when they were onset, body, or phoneme units. Additionally, both ML and matched controls were much better able to find words in an anagrams task (Treiman & Chafetz, 1987) when they resulted from the combination of segments corresponding to syllables than when they did not. It is suggested that the relationship between print and sound is represented at multiple levels (including the syllable) (Shallice, Warrington, & McCarthy, 1983) and that ML's nonword reading impairment is the result of disruption of representations below the level of the syllable. PMID- 9854444 TI - E-mail, snail mail, phone, or fax. Reporting adverse reactions and other product problems. PMID- 9854445 TI - MedWatch. FDA's 'heads up' on medical product safety. PMID- 9854446 TI - Staking a claim to good health. FDA and science stand behind health claims on foods. PMID- 9854447 TI - Miracle drugs vs. superbugs. Preserving the usefulness of antibiotics. PMID- 9854448 TI - Sizing up surgery. PMID- 9854449 TI - Convictions upheld in tainted animal feed case. PMID- 9854450 TI - Crab company cleans house. PMID- 9854451 TI - Doctor who took her patients' morphine for herself sentenced under anti-tampering law. PMID- 9854452 TI - Anti-inflammatory actions of glucocorticoids: molecular mechanisms. AB - 1. Glucocorticoids are widely used for the suppression of inflammation in chronic inflammatory diseases such as asthma, rheumatoid arthritis, inflammatory bowel disease and autoimmune diseases, all of which are associated with increased expression of inflammatory genes. The molecular mechanisms involved in this anti inflammatory action of glucocorticoids is discussed, particularly in asthma, which accounts for the highest clinical use of these agents. 2. Glucocorticoids bind to glucocorticoid receptors in the cytoplasm which then dimerize and translocate to the nucleus, where they bind to glucocorticoid response elements (GRE) on glucocorticoid-responsive genes, resulting in increased transcription. Glucocorticoids may increase the transcription of genes coding for anti inflammatory proteins, including lipocortin-1, interleukin-10, interleukin-1 receptor antagonist and neutral endopeptidase, but this is unlikely to account for all of the widespread anti-inflammatory actions of glucocorticoids. 3. The most striking effect of glucocorticoids is to inhibit the expression of multiple inflammatory genes (cytokines, enzymes, receptors and adhesion molecules). This cannot be due to a direct interaction between glucocorticoid receptors and GRE, as these binding sites are absent from the promoter regions of most inflammatory genes. It is more likely to be due to a direct inhibitory interaction between activated glucocorticoid receptors and activated transcription factors, such as nuclear factor-kappa B and activator protein-1, which regulate the inflammatory gene expression. 4. It is increasingly recognized that glucocorticoids change the chromatin structure. Glucocorticoid receptors also interact with CREB-binding protein (CBP), which acts as a co-activator of transcription, binding several other transcription factors that compete for binding sites on this molecule. Increased transcription is associated with uncoiling of DNA wound around histone and this is secondary to acetylation of the histone residues by the enzymic action of CBP. Glucocorticoids may lead to deacetylation of histone, resulting in tighter coiling of DNA and reduced access of transcription factors to their binding sites, thereby suppressing gene expression. 5. Rarely patients with chronic inflammatory diseases fail to respond to glucocorticoids, although endocrine function of steroids is preserved. This may be due to excessive formation of activator protein-1 at the inflammatory site, which consumes activated glucocorticoid receptors so that they are not available for suppressing inflammatory genes. 6. This new understanding of glucocorticoid mechanisms may lead to the development of novel steroids with less risk of side effects (which are due to the endocrine and metabolic actions of steroids). 'Dissociated' steroids which are more active in transrepression (interaction with transcription factors) than transactivation (GRE binding) have now been developed. Some of the transcription factors that are inhibited by glucocorticoid, such as nuclear factor-kappa B, are also targets for novel anti-inflammatory therapies. PMID- 9854453 TI - Effects of beta-blockers on the concentration and oxidizability of plasma lipids. AB - 1. beta-blockers improve morbidity and mortality after myocardial infarction, probably by several mechanisms. We investigated potentially relevant effects of beta-blockers in vivo and in vitro on plasma lipid oxidizability. Forty-two healthy men were randomized to receive placebo (13), metoprolol (14) or propranolol (15). 2. At 4 weeks, the effects on heart rate, blood pressure and lipids appeared similar and subjects taking a beta-blocker were combined. Compared with placebo, those on a beta-blocker gained 0.5 kg in weight (P = 0.04), heart rate fell from 63 to 52 beats/min (P < 0.0001) and blood pressure fell from 116/74 to 113/69 mmHg (P < 0.005); high-density lipoprotein (HDL) cholesterol fell from 1.26 to 1.11 mmol/l (P = 0.005), there being no change in the ratio of free to esterified cholesterol in HDL, and there was an apparent rise in serum triacylglycerols from 1.18 to 1.43 mmol/l (P = 0.15 when adjusted for weight gain). Low-density lipoprotein (LDL)-cholesterol and lipoprotein (a) did not change. In this study, the oxidizability of LDL was unaffected by beta blocker therapy. beta-blockade was not associated with any change in LDL fatty acid profile, or beta-carotene or alpha-tocopherol content which might account for the reduced LDL oxidizability previously reported in patients treated with beta-blockers. Furthermore, neither atenolol nor propranolol, at concentrations up to 100 mumol/l, had any effect on in vitro oxidizability of LDL obtained from healthy volunteers. 3. In contrast to the favourable haemodynamic effects conferred by beta-blockers, the effects on weight and serum triacylglycerols and HDL-cholesterol appear to be adverse and we did not demonstrate any changes in lipid oxidizability which might be relevant to the protective effects of beta blockers in patients with coronary disease. PMID- 9854454 TI - Elevated heart rate variability in physically active young and older adult women. AB - 1. Low heart rate variability is associated with an increased risk of cardiac sudden death, coronary heart disease and all-cause mortality. We have previously shown that physically active postmenopausal women demonstrate higher levels of heart rate variability and cardiac baroreflex sensitivity compared to their sedentary peers. The purpose of the present prospective study was to test the hypothesis that heart rate variability and cardiac baroreflex sensitivity would be reduced with age in sedentary but not physically active women. To accomplish this, we measured heart rate variability (both time and frequency domain) and spontaneous cardiac baroreflex sensitivity (SBRS, sequence method) in the sitting posture in 23 sedentary women [11 premenopausal and 12 postmenopausal (age, 28 +/ 1 and 61 +/- 2 years; VO2max, 35.3 +/- 1.4 and 21.7 +/- 1.5 ml.min-1.kg-1 respectively] and in 22 physically active women [12 premenopausal and 10 postmenopausal (age, 31 +/- 1 and 59 +/- 2 years; VO2max, 52.5 +/- 1.4 and 39.7 +/- 1.8 ml.min-1.kg-1)]. 2. The S.D. of the R-R interval (time domain) was reduced (P < 0.05) with age in both sedentary (52 +/- 6 versus 33 +/- 4 ms) and physically active women (72 +/- 8 versus 49 +/- 9 ms). The high-frequency power (3740 +/- 1527 versus 915 +/- 188 and 9516 +/- 2849 versus 2803 +/- 1083 ms2/Hz), total power of heart rate variability and SBRS (11 +/- 2 versus 7 +/- 2 and 19 +/ 3 versus 13 +/- 2 ms/mmHg) also demonstrated similar age-related reductions in sedentary and physically active women, respectively (all P < 0.05). The S.D. of the R-R interval, high-frequency and total power of heart rate variability, and SBRS were higher (all P < 0.05) in the physically active compared with the sedentary women at any age. There was no significant influence of age or physical activity status on the low-frequency power of heart rate variability. In addition, no significant differences in any of the time or frequency domain measures of heart rate variability or SBRS were observed in users compared with non-users of hormone replacement therapy. 3. The results of the present study suggest that heart rate variability and cardiac baroreflex sensitivity decline similarly with age in healthy sedentary and physically active women. However, physically active women demonstrate higher levels of heart rate variability and cardiac baroreflex sensitivity compared with their sedentary peers, regardless of age. PMID- 9854455 TI - Increased plasma adrenomedullin concentrations during cardiac surgery. AB - 1. Adrenomedullin (AM), a potent hypotensive peptide, was originally isolated from human phaeochromocytoma. Plasma AM concentrations are elevated in hypertension, heart failure and renal failure in proportion to the severity of the disease. This study was performed to investigate the pathophysiological significance of AM during cardiac surgery. 2. Serial blood samples were obtained from patients undergoing cardiac surgery and plasma AM concentrations were determined by specific radioimmunoassay. 3. Plasma AM concentrations did not increase with anaesthesia or surgery (n = 9). Plasma AM concentrations gradually increased during cardiopulmonary bypass and after pulmonary reperfusion. After pulmonary reperfusion, plasma AM concentrations increased further. In addition, we measured plasma AM concentrations in the pulmonary vein (n = 8) and coronary sinus (n = 8) to examine the contribution of the lungs and heart to the increase in circulating AM concentrations after cardiopulmonary bypass. However, no significant differences were seen in plasma AM concentrations of the pulmonary vein or the coronary sinus and the aorta. Peak AM concentrations during cardiac surgery correlated with duration of surgery. Elevated plasma AM levels during and after surgery began to decline next day after surgery and returned to normal levels 7 days after surgery. 4. These results demonstrate that plasma AM concentrations increase during cardiac surgery and that the duration of surgery may be related to the changes in AM concentrations. Taken together with recent findings that vascular endothelial cells and vascular smooth muscle cells actively produce AM, these results suggest that plasma AM during cardiac surgery may act as a vasodilatory hormone. PMID- 9854456 TI - How does treatment influence endocrine mechanisms in acute severe heart failure? Effects on cardiac natriuretic peptides, the renin system, neuropeptide Y and catecholamines. AB - 1. Hormones involved in cardiovascular regulation are influenced by drug treatment. It is therefore difficult to study endocrine mechanisms in heart failure as most patients are already on treatment by the time they reach hospital. 2. We studied nine hospital in-patients before and after treatment of acute New York Heart Association class IV heart failure. 3. Before treatment, plasma brain and atrial natriuretic peptides were markedly elevated (BNP 121 +/- 26 pg/ml, ANP 163 +/- 33 pg/ml; normal range: BNP 3.9 +/- 0.3 pg/ml, ANP 8.6 +/- 0.8 pg/ml) and correlated positively with serum creatinine and left ventricular end-diastolic diameter and negatively with ejection fraction. Eight patients improved and one died. 4. With improvement plasma ANP and BNP fell. Initial renin activity was within the normal range but increased on treatment. Plasma neuropeptide Y and adrenaline remained normal before and after treatment in the eight patients who improved. Initial plasma noradrenaline was in the normal range in four of these patients and just above normal in a further four. In the patient who died, initial plasma neuropeptide Y and catecholamines were very high. 5. Plasma BNP emerged as complementary to ANP as a dynamic index in severe heart failure; however, renal function is also an important determinant of plasma BNP and ANP. There is little evidence for activation of the circulating renin angiotensin-aldosterone system or neuropeptide Y before treatment of acute heart failure. PMID- 9854457 TI - Acute hypertension impairs endothelium-dependent vasodilation. AB - 1. Previous investigations have demonstrated an impaired endothelium-dependent vasodilatation (EDV) in patients with hypertension. The present study aimed to investigate if an acute rise in blood pressure to hypertensive levels impairs EDV in otherwise normotensive subjects. 2. Twenty-seven young, healthy, normotensive subjects were studied. Eight of these underwent evaluation of EDV and endothelium independent vasodilatation (EIDV) by means of forearm blood flow measurements during local intra-arterial infusions of methacholine (2 and 4 micrograms/min) and sodium nitroprusside (5 and 10 micrograms/min), before and after 1 h of sustained hypertension, induced by noradrenaline given intravenously. Identical measurements were made in 11 subjects before and during concomitant local intra arterial infusion of noradrenaline without change in blood pressure and eight subjects were studied during saline infusions. 3. One hour of sustained hypertension (diastolic blood pressure > 95 mmHg) significantly attenuated both forearm blood flow (17.4 +/- 6.8 versus 27.4 +/- 6.8 ml.min-1.100 ml-1 tissue at baseline, P < 0.05) and forearm vascular resistance decrease (3.2 +/- 0.87 versus 7.4 +/- 2.5 units at baseline, P < 0.05) during methacholine infusion. These attenuations were significantly more pronounced for methacholine than for sodium nitroprusside (P < 0.05). In contrast, local intra-arterial noradrenaline infusions impaired vasodilatation induced by methacholine and sodium nitroprusside to a similar extent. Saline infusions did not change either EDV or EIDV. 4. Thus, an acute rise in blood pressure to hypertensive levels induced by noradrenaline impaired EDV more than EIDV in otherwise normotensive subjects, while no such selective effect of local noradrenaline was seen, suggesting that a high blood pressure impairs endothelial vasodilator function. PMID- 9854458 TI - Suppression of post-glucose hyperinsulinaemia does not affect blood pressure in either normotensive or hypertensive subjects. AB - 1. Hyperinsulinaemia and insulin resistance are thought to be intimately involved in the development of hypertension, but controversy remains as to whether hyperinsulinaemia is a consequence or a cause of hypertension per se, and whether it plays a role in the short-term regulation of blood pressure. 2. We studied six hypertensive patients [blood pressure 161(9)/101(2) mmHg] and seven normotensive control subjects [blood pressure 122(6)/76(4) mmHg], (P < 0.005) using two oral glucose tolerance tests of 3 h duration. In one of these tests the endogenous insulin response was inhibited with subcutaneous octreotide. 3. After placebo, hypertensive patients had slightly but significantly higher blood glucose levels than controls (P < 0.0001), but comparable insulin concentrations (P > 0.5). Plasma noradrenaline levels were consistently lower in the hypertensive group (P < 0.001). Blood pressure did not change in either group during the 3 h after glucose ingestion. 4. Octreotide completely abolished the immediate insulin response to glucose in all subjects (both P < 0.0001) and caused a delayed and significantly increased glycaemic response in both groups (P < 0.0001). There were no significant differences in plasma glucose responses between groups: however, after octreotide, the hypertensive subjects had a greater insulin suppression than the controls (P < 0.02). Octreotide suppressed noradrenaline levels in the normotensive group (P < 0.001); they were also suppressed in the hypertensive group, but just failed to reach significance (P = 0.056). Throughout the study the hypertensive group's noradrenaline levels remained generally lower than those in the control group (P < 0.0001). 5. In this study there were no differences between hypertensive and normotensive subjects in fasting or post glucose insulin levels, nor any significant change in blood pressure in either group when post-glucose hyperinsulinaemia was suppressed. This argues against insulin playing a direct role in the short-term regulation of blood pressure. PMID- 9854459 TI - Cardiovascular autonomic testing in adolescents with type I (insulin-dependent) diabetes mellitus: an 18-month follow-up study. AB - 1. Autonomic abnormalities are frequent in adult patients with diabetes mellitus and progress slowly; little is known about frequency and progression of autonomic abnormalities in childhood. 2. To assess whether autonomic abnormalities are already present in childhood, we evaluated the cardiovascular reflexes, the spectral analysis of spontaneous fluctuations in RR interval and blood pressure (low- and high-frequency), and the baroreflex sensitivity at rest, and after vagal (controlled breathing) and sympathetic activation (tilting) in 25 adolescents with Type I diabetes mellitus, aged 10-17 years, at baseline and after 18 months follow-up, and in 20 age- and sex-matched controls. 3. Cardiovascular reflexes were similar in both patients and controls. Similar significant changes in percentage low- and high-frequency (P < 0.005) from rest to tilting and to control breathing were observed in both patients and controls. The baroreflex sensitivity was also similar in patients and controls. Mild and non-systematic correlations were observed between autonomic tests and disease duration or metabolic control. After 18 months follow-up no changes were observed in any of the measured variables. Correlations with metabolic control remained unchanged. 4. These results indicate a substantial stability of cardiovascular autonomic function in childhood diabetes, and suggest that autonomic abnormalities are likely to develop at an older age. PMID- 9854460 TI - Increased oxidative stress in rat liver and pancreas during progression of streptozotocin-induced diabetes. AB - 1. Oxygen free radicals have been suggested to be a contributory factor in complications of diabetes mellitus. There are many reports indicating the changes in parameters of oxidative stress in diabetes mellitus. In this study we aimed to identify whether oxidative stress occurs in the liver and pancreas in the initial stages of development of diabetes. 2. We therefore investigated the lipid peroxide level (thiobarbituric acid-reactive substances, TBARS) and activities of antioxidant enzymes [superoxide dismutase (SOD), catalase and glutathione peroxidase] in liver and pancreas of control and streptozotocin-induced diabetic rats at various stages of development of diabetes. 3. Male Sprague-Dawley rats were divided into two groups: group I, control (n = 42) and group II, diabetic (n = 42). Each group was further subdivided into seven groups consisting of six rats each. Rats in these subgroups were studied at weekly intervals (0 to 6 weeks). Plasma glucose levels, TBARS levels and activities of antioxidant enzymes were measured in liver and pancreas at various time intervals. 4. There was a significant (P < 0.05) and progressive increase in TBARS levels of liver and pancreas in the diabetic group. Total SOD and Cu-Zn-SOD activity increased (P < 0.05) with progression of diabetes while Mn-SOD activity showed no significant change in either tissue. Catalase and glutathione peroxidase activities increased significantly (P < 0.05) in liver and pancreas. 5. Immunohistochemical study of pancreatic islet revealed a decrease in the expression of insulin with progression of diabetes. However, glucagon and somatostatin showed an increase in immunoreactivity and a difference in their distribution pattern. 6. The findings of the present study suggest that oxidative stress starts at early onset of diabetes mellitus and increases progressively. In conclusion, the structural damage to these tissues or complications of diabetes mellitus may be due to oxidative stress. PMID- 9854461 TI - Serum leptin level: possible association with haematopoiesis in adolescents, independent of body mass index and serum insulin. AB - 1. The obese gene product leptin, secreted exclusively from adipocytes, was discovered to serve as a satiety factor and to play an important role in regulating body weight. In adults, the serum leptin level reportedly increases with the degree of obesity. Leptin receptors are expressed in various tissues, and recent in vitro studies suggest a role for leptin in haematopoiesis. 2. The present study was designed to clarify the relationship between serum leptin and body mass index, peripheral blood cell counts, serum cholesterol, high-density lipoprotein-cholesterol, insulin and cortisol levels in 299 Japanese male adolescents aged 15-16 years. 3. With simple linear correlation, log [serum leptin] showed a strong correlation with body mass index (r = 0.56), log [insulin] (r = 0.36) and leucocyte count (r = 0.22) (P < 0.001 for all). There were also correlations with systolic blood pressure, erythrocyte count, haematocrit and high-density lipoprotein-cholesterol (P < 0.01 for all). Even after adjustment for body mass index and log [insulin], log [leptin] correlated with leucocyte (P = 0.004) and erythrocyte (P = 0.057) counts. Stepwise multiple regression analyses revealed log [leptin] to correlate significantly with body mass index, log [insulin] and the leucocyte count (P < 0.005 for all, r2 = 0.399). 4. To our knowledge, this is the first clinical study to show the possible association of serum leptin level with blood cell counts, independent of body mass index and serum insulin. We conclude that these data further support a role for leptin in haematopoiesis. PMID- 9854463 TI - Effect of long-term octreotide and isosorbide dinitrate on haemodynamics in rats with portal vein stenosis. AB - 1. Both octreotide and isosorbide dinitrate have been shown to have portal hypotensive effects in animals with portal hypertension. Moreover, in both animals and humans with portal hypertension, the reduction of portal pressure was enhanced when nitrovasodilators were combined with propranolol or vasopressin. The present study was undertaken to evaluate the effect of long-term administration of octreotide and isosorbide dinitrate on haemodynamics in rats with portal vein stenosis. 2. Portal hypertension was induced by portal vein stenosis. Portal hypertensive rats were allocated into one of four groups (eight rats in each group): vehicle group, octreotide group (100 micrograms/kg via subcutaneous injection every 12 h), isosorbide dinitrate group (5 mg/kg via gastric gavage every 12 h) and combined treatment group. Drug was given for eight consecutive days, starting 1 day before surgery. Haemodynamic values were measured using a radioactive microsphere technique. 3. Long-term octreotide treatment decreased portal pressure and improved the hyperdynamic circulation. In contrast, long-term administration of isosorbide dinitrate reduced portal pressure but did not ameliorate vasodilatation. A combination of octreotide and isosorbide dinitrate improved the hyperdynamic circulation with a reduction of portal pressure. In addition, the mean value of portal pressure after combination treatment was significantly lower than in rats receiving octreotide alone. 4. These results showed that, in rats with portal hypertension, long-term combined administration of octreotide and isosorbide dinitrate improved the hyperdynamic circulation together with a more profound reduction of portal pressure than rats receiving octreotide alone. PMID- 9854462 TI - Increased nitric oxide synthesis and inducible nitric oxide synthase expression in patients with alcoholic and non-alcoholic liver cirrhosis. AB - 1. The synthesis and release of nitric oxide may play a role in the pathogenesis of peripheral vasodilatation and hyperdynamic circulation observed in liver cirrhosis. In this work, we analysed the synthesis of nitric oxide by the lympho mononuclear cells of peripheral blood from patients with chronic alcoholic and non-alcoholic liver disease and we identified the isoform of nitric oxide synthase involved in the increased nitric oxide synthesis. 2. Patients were classified following clinical and histological criteria in non-alcoholic cirrhotic, alcoholic cirrhotic and non-cirrhotic chronic liver disease. We studied clinical and analytical characteristics, haemodynamic parameters and endotoxin levels in these patients. 3. Cirrhotic patients showed an increase of cardiac output and a decrease of peripheral vascular resistance. These patients had higher levels of plasma endotoxin than those observed in the control group. N omega-Nitro-L-arginine methyl ester (L-NAME)-inhibitable nitrite production from mononuclear lymphocyte cells was higher in patients than in the control group, the highest levels being in non-alcoholic cirrhotic patients, and the lowest levels in patients with non-cirrhotic alcoholic liver disease. 4. Immunocytochemistry studies revealed a positive immunoreactivity for the inducible isoform of nitric oxide synthase in lympho-mononuclear cells that was more evident in non-alcoholic than in alcoholic cirrhotic patients. By Northern blot, inducible nitric oxide synthase mRNA expression was observed only in lymphomononuclear cells from non-alcoholic cirrhotic patients. 5. Our patients show a correlation between nitric oxide synthesis, endotoxin levels and haemodynamic parameters. 6. These findings indicate that lympho-mononuclear cell stimulation may play a role in elevated nitric oxide production in hepatic cirrhosis. Thus, this increased nitric oxide synthesis could be implicated in the pathogenesis of the haemodynamic disturbances frequently found in cirrhotic patients. This increase seems to be induced, at least in part, by activation of an inducible isoform of nitric oxide synthase. PMID- 9854464 TI - Deep venous thrombosis: a prospective 3-month follow-up using duplex scanning and strain-gauge plethysmography. AB - 1. The purpose of the study was to evaluate the degree of thrombus regression, development of valvular insufficiency, impaired calf muscle pump function and clinical symptoms after a period of acute deep venous thrombosis. 2. Seventy patients with acute deep venous thrombosis, diagnosed by duplex scanning or venography, received treatment with heparin and oral coumarin derivatives according to a standard protocol. All patients wore graduated compression stockings during the whole study period. Duplex scanning was performed at diagnosis and 1 and 3 months later to measure thrombus mass and reflux. The supine venous pump function test was used to assess calf muscle pump function. 3. Three months follow-up was completed in 60 patients. In total 218 (28%) out of 780 vein segments were initially thrombosed and 134 (17%) could not be traced. A statistically significant reduction of thrombus mass was recorded throughout the study period. Total resolution of thrombosis in all vein segments occurred in 25% of the patients within 1 month and in 40% in 3 months. There was no difference in regression between the various proximal vein segments. Distal segments showed more regression than proximal segments. Reflux occurred in 27% of the initially thrombosed veins and in 15% of the patent veins. Patients who showed total resolution after 1 month had a significantly higher calf muscle pump function than patients without total resolution (70%pf vs 61%pf, P < 0.05). Patients with reflux in two or more segments had a significantly lower calf muscle pump function than patients with reflux in less than two segments (58%pf vs 69%pf, P < 0.05). 4. Using duplex scanning and the supine venous pump function test 3 months after an acute deep venous thrombosis, overall haemodynamic abnormalities and local site of valve incompetence could readily be identified. Patients with haemodynamic abnormalities might be at risk to develop the post-thrombotic syndrome. There was no difference in thrombus regression between the various proximal vein segments. Reflux was significantly more often seen in initially thrombosed veins. Thirty-five percent of the patients developed an abnormal calf muscle pump function after 3 months. Patients with early resolution of thrombus had a higher calf muscle pump function after 3 months. PMID- 9854465 TI - Colour Doppler sonographic diagnosis of upper limb venous thromboses. AB - 1. Upper limb venous thromboses are considered to be a rare event, but in large hospitals with a lot of patients who receive aggressive intravenous therapy the number of thromboses seem to increase. 2. We have analysed all the cases of upper limb venous thrombosis which occurred at the Essen University Hospital between the years of 1992-1996. All patients were examined using colour Doppler sonography. 3. Out of 827 patients that were examined, a thrombosis was diagnosed in 334 cases. The subclavian vein was involved in 69% of all thromboses. Isolated jugular vein thrombosis was found in 17% of the thromboses, combined thromboses of the jugular and subclavian vein in 19%. In 182 cases the patients were treated for primarily a malignant illness. In 96 cases we found an association with venous port-systems or central venous catheters. 4. More than 40,000 patients a year were treated at the university hospital. Considering this huge number of patients the thrombosis of the upper limb is still rare. The use of colour Doppler sonography allows an early and safe diagnosis of the thrombosis without straining the patients. PMID- 9854466 TI - Concentration-dependent stimulation of intestinal phase III of migrating motor complex by circulating serotonin in humans. AB - 1. The influence of circulating 5-hydroxytryptamine (serotonin) on small intestinal motility was investigated in healthy volunteers. 2. Small intestinal motility was studied by means of a constantly perfused multi-channel manometry tube, connected to a computer system. 3. Intravenous infusions of either 5 hydroxytryptamine at increasing doses or saline were given over a period of 4 h. 4. 5-Hydroxytryptamine infusion dose-dependently increased plasma 5 hydroxytryptamine from approximately 2 to 10 and 25 nmol/l respectively, as well as urinary excretions of 5-hydroxytryptamine and 5-hydroxyindole acetic acid, a major 5-hydroxytryptamine metabolite. 5. The number of phase III of the migrating motor complex originating in the small intestine was dose-dependently increased by 5-hydroxytryptamine, and found to correlate to the plasma concentration of 5 hydroxytryptamine. The fraction of phase III also increased at the expense of phase II activity. In addition, 5-hydroxytryptamine increased the motility index, propagation velocity of phase III activity and the amplitude of contractions during phase III. 6. Whereas the low dose of 5-hydroxytryptamine (15 nmol.min 1.kg-1) had no haemodynamic effects, an increase in heart rate by approximately 20 beats/min, without change in blood pressure, was observed at the higher dose (60 nmol.min-1.kg-1). Respiratory parameters did not change during infusion of 5 hydroxytrytamine at either dose. 7. In conclusion, elevation of circulating 5 hydroxytryptamine by intravenous infusion results in more frequent and faster propagating migrating motor complexes in the human small intestine during the inter-digestive period. PMID- 9854467 TI - Effect of topical butyrate on rectal epithelial kinetics and mucosal enzyme activities. AB - 1. This study aimed to determine the effect of luminal butyrate on proliferative kinetics, a differentiation marker (alkaline phosphatase), and a molecule that controls cell-substratum adhesion (urokinase) in histologically normal human rectal mucosa. 2. Ten subjects with a colonoscopically normal colon (seven had previous adenomas) were given either butyrate or saline enemas for 4 days in a double-blind cross-over manner. Rectal biopsies were taken before and after each course of enemas. Epithelial proliferative kinetics were measured immunohistochemically using antibodies to proliferating cell nuclear antigen. Urokinase and alkaline phosphatase activities were measured spectrophotometrically in biopsy homogenates. 3. Both saline and butyrate enemas were well tolerated and induced no histological change except for a significant increase in crypt length (P < 0.05). The number of proliferating cells per crypt also increased significantly after butyrate (P = 0.018). 4. Compared with saline enemas, butyrate did not affect kinetic indices nor alkaline phosphatase activities. However, mucosal urokinase activities were significantly lower in butyrate-treated patients (9.5 +/- 2.0 i.u./g) than in saline-treated patients (12.8 +/- 2.0 i.u./g; P = 0.045). 5. Delivering of extra butyrate to the distal colon in healthy subjects may stabilize cell-substratum adhesion in surface epithelium and therefore offer a potential mechanism by which elevating distal colonic luminal butyrate concentrations might be beneficial in patients with colitis or hyperproliferative large bowel epithelium. PMID- 9854468 TI - Regional differences in protein carboxymethylation in post-mortem human brain. AB - 1. The aim of this study was to determine the pattern of protein carboxymethylation in different regions of the human brain. 2. The availability of protein methylation sites was determined by measurement of the incorporation of methyl-3H groups into proteins isolated from post-mortem brain tissue. The stability of protein carboxymethylation in post-mortem brain was determined by sampling post-mortem pig and human brain tissue at intervals during the first 24 h after death. This method has previously been used to demonstrate that decreased protein carboxymethylation occurred in post-mortem pig brain when methionine synthase was inhibited. 3. There were no significant differences in the protein carboxymethyltransferase activity in samples of pig brain obtained at the time of death compared with that obtained when the same tissue was maintained at room temperature for up to 24 h after death. Similarly, there were no significant differences in the protein carboxymethyltransferase activity in samples isolated from human brain 12 h after death compared with that obtained from the same human brain tissue maintained at room temperature for up to 24 h after death. These results suggest that the level of carboxymethylation of proteins from human post mortem brain obtained within 24 h of death is not significantly different to the level present at the time of death. To characterize the distribution of protein carboxymethylation in human brain, nine regions of post-mortem brain were sampled from 16 human subjects. Protein carboxymethyltransferase activity was lowest in the cerebellum (P < 0.05) and highest in cortical white matter compared with other regions of the brain (P < 0.05). No significant differences in protein carboxymethyltransferase activity were noted between other regions of the cortex or the subcortical regions. 4. In human cortical white matter there are more available sites for protein carboxymethylation than other brain regions. This may explain the greater sensitivity of white matter to the adverse consequences of hypomethylation associated with vitamin B12 deficiency. Post-mortem brain tissue can serve as a tool for the study of physiological or pathological factors which influence human brain protein methylation in vivo. PMID- 9854469 TI - Anti-infective catheters: novel strategies to prevent nosocomial infections in oncology. AB - Intravenous access contributes significantly to the therapeutical success and to the comfort of oncologic patients. The highest risk for bloodstream infections, however, is vascular catheter-mediated. In oncology high mortality is associated with Pseudomonas aeruginosa, Candida albicans and Staphylococcus aureus sepsis. Besides established hygienic measures, the coupling or incorporation of antimicrobial substances to or into catheter materials may be a suitable way to prevent the development of catheter-associated infections. Here we present a risk benefit evaluation of different models of antimicrobial catheter coated with silver, antiseptics or antibiotics. The controversial reports on clinical efficacy and the potential of adverse reactions due to silver and antiseptic coated catheters are discussed. The microbiological, pharmaceutical and physicochemical backgrounds of different types of coating are discussed in detail. Incorporation of antimicrobial agents into long-term silicon catheters providing a slow release of those substances through the external and internal surfaces of catheters may be the most effective technological innovation for reducing biomaterial-mediated nosocomial infections. PMID- 9854470 TI - Expression of MAGE-1, MAGE-2 and MAGE-3 genes in human gastric carcinomas; lack of evidence for cytotoxic effects in cases with simultaneous expression of MAGE-3 and HLA-A2. AB - The human MAGE gene products are recognized by major histocompatibility complex restricted cytotoxic T lymphocytes. We analyzed by RT-PCR the expression of MAGE 1, MAGE-2, MAGE-3 and HLA-A2 genes in 10 human gastric cancer cell lines and 46 human stomachs removed due to advanced gastric carcinomas. All the cell lines expressed MAGE genes, except for MKN-45 and -74 which lacked the expression of MAGE-1 and -3 genes in this study. Of the 46 gastric carcinomas, MAGE-1, -2 and 3 genes were expressed in 14 (30%), 10 (22%) and 26 (57%) cases, respectively, regardless of histological type. Normal gastric mucosa and intestinal metaplastic mucosa showed no expression of these genes. HLA-A2 gene expression was noted in 14 both normal and carcinoma cases. Simultaneous expression of MAGE-3 and HLA-A2 genes was noted in 7 cases. Mean apoptotic and Ki-67-labeling indices (AI and KI) of carcinoma cells were 2.3 +/- 0.5 and 48.1 +/- 6.0 in 7 cases, and 2.8 +/- 0.2 and 47.3 +/- 2.7 in the other 39 cases lacking the expression of MAGE-3 and/or HLA-A2 genes, respectively. The two-year survival rate did not differ between the two groups. Although this study confirmed the relatively higher expression of the MAGE gene family in human advanced gastric carcinomas, it might suggest that simultaneous expression of MAGE-3 and HLA-A2 genes does not necessarily imply the induction of cancer cell apoptosis by CTL. PMID- 9854472 TI - Caffeine-potentiated chemotherapy and conservative surgery for high-grade soft tissue sarcoma. AB - We report here the results of preoperative and postoperative caffeine-potentiated chemotherapy and limb-sparing surgery for soft-tissue sarcomas. Thirty-six patients with histologically high-grade soft-tissue sarcomas were treated with caffeine-potentiated chemotherapy and conservative surgery (25 cases of limb sparing surgery and 11 of local tumor excision). There were 13 patients with malignant fibrous histiocytoma (MFH), eight with synovial sarcoma, five with liposarcoma, four with malignant schwannoma, four with epithelioid sarcoma, one with leiomyosarcoma and one with extraskeletal chondrosarcoma. Nine patients were at stage III with lung metastasis and the other 27 at stage IIB without metastasis; 22 were male and 14 female with a mean age of 48 years, ranging from 16 to 77. For intra-arterial preoperative chemotherapy, we administered 2-5 courses of cisplatin (120 mg/m2), doxorubicin (30 mg/m2 x 2 days), and caffeine (1.5 g/m2 x 3 days) to 18 patients, and cisplatin and caffeine to the other 18. Although 15 patients had already undergone unplanned tumor excision at other hospitals before preoperative chemotherapy, all patients underwent definitive limb-sparing surgery after the preoperative chemotherapy. Surgical margins were wide for 28 patients, marginal for three and intralesional for five. Local tumor recurrence was seen in one patient with MFH and one with epithelioid sarcoma. Of the 27 stage IIB patients, lung metastasis newly developed in one with MFH, three with synovial sarcoma, two with malignant schwannoma and one with leiomyosarcoma. As for the effects of preoperative chemotherapy in the 33 eligible cases, radiographically confirmed complete response was seen in two patients, partial response in 20 and no response in 11. Histological response to this preoperative chemotherapy consisted of grade I (no response) in 14, grade II (50-90% necrosis) in four, grade III (> 90% necrosis) in eight, and grade IV (no viable cells) in seven cases. An overall objective response rate of 73% was obtained. With the mean follow-up period of 58 months (5-101 months), the overall 5-year cumulative survival rate ascertained with the Kaplan-Meier method was 63% and that of stage II patients 81%. Eight of the nine stage III patients died of metastatic disease within two and a half years from the beginning of the treatment. In conclusion, caffeine-potentiated chemotherapy and limb-sparing surgery brought good results for stage II nonmetastatic soft-tissue sarcomas. The problem of treatment for stage III metastatic soft-tissue sarcomas, however, remains unsolved. PMID- 9854471 TI - Preneoplastic oral lesions: the clinical value of image cytometry DNA analysis, p53 and p21/WAF1 expression. AB - BACKGROUND: Various mucosal lesions are frequently encountered in the oral cavity. Neither macroscopic nor microscopic evaluation of these lesions gives any reliable information concerning the risk of cancer development. MATERIAL AND METHODS: From 21 patients, 29 mucosal lesions were found to precede development of invasive squamous cell carcinoma or carcinoma in situ at the same location. The lesions were matched to 29 control lesions, with the same grade of dysplasia and from exactly the same locations but without subsequent cancer during a mean follow up of 112 months (46-194). The specimens were evaluated using Image Cytometry DNA analysis and immunohistochemical analysis of p53 and p21/WAF1 expression. RESULTS: Lesions prior to carcinomatous development displayed a higher degree of DNA aberration as compared with the control lesions. p53 and p21/WAF1 evaluation did not reveal any differences between cases and controls. CONCLUSION: Image Cytometry DNA analysis is an useful adjunct to histopathological evaluation of oral mucosal lesions for prediction of risk of malignant transformation. PMID- 9854473 TI - Serum hepatocyte growth factor level associate with gastric cancer progression. AB - We have measured serum HGF levels from 80 gastric cancer patients and 51 normal subjects by enzyme-linked immunosorbent assay. The results showed that the mean value of serum HGF level in gastric cancer patients was significantly higher than in normal subjects (0.30 +/- 0.02 vs 0.22 +/- 0.05 ng/ml; p = 0.005). The increase was stage related. Patients with serum HGF < or = 0.30 ng/ml survived longer than those with serum HGF > 0.30 ng/ml (p = 0.02). These data suggest that HGF involve in progression of gastric cancer. PMID- 9854474 TI - The calcium-binding protein calretinin-22k is detectable in the serum and specific cells of cancer patients. AB - BACKGROUND: Calretinin-22k (CR-22k), an alternatively spliced form of calretinin (CR) belongs to the EF-hand family of calcium-binding proteins and is expressed in several colon adenocarcinoma cell lines (e.g. WiDr, HT-29). MATERIALS AND METHODS: Serum samples of cancer patients were screened with a sandwich ELISA technique using the CR-specific antiserum 7696. Highly positive samples were analyzed by Western blots and immunohistochemistry. RESULTS: CR-22k was detected in the serum of several patients and values were as high as 0.19 microgram/ml. Western blot analysis confirmed the identity of the bound protein as CR-22k. The highest concentrations were detected in patients with colon or breast cancer, but also in a patient with ischemic necrosis of the gut. CR immunoreactivity was localized to epithelial cells, nerve fibres, cells of the connective tissue and to mesothelial cells. CONCLUSIONS: Our results establish that CR-22k is detectable in the serum of cancer patients under specific pathological conditions. PMID- 9854475 TI - Immunohistochemical localization of epithelial glycoprotein EGP-2 and carcinoembryonic antigen in normal colonic mucosa and colorectal tumors. AB - Epithelial glycoprotein EGP-2 and carcinoembryonic antigen (CEA) are transmembrane glycoproteins and cell surface markers. Eighty-four colorectal tumors including 23 adenomas (2 mild, 13 moderate, and 8 severe atypia) and 61 adenocarcinomas (33 well- and 28 moderately differentiated) as well as adjacent normal colonic mucosa (51 cases) were studied for the immunolocalization of EGP-2 as detected by the monoclonal antibody VU-1D9, and compared with CEA expression. In the normal colonic mucosa, basolateral VU-1D9 expression in the surface epithelial cells was constantly seen in all 51 cases, while weak apical CEA staining in the surface epithelium was seen in 25% (13/51) of the cases. In 91% (21/23) of the adenomas, regardless of the grade of atypia, VU-1D9 labeled the basolateral membrane of a few surface lining cells leaving atypically proliferating glands negative, while CEA expressed strong apical staining in the surface epithelial cells as well as atypically proliferating glands. The well differentiated adenocarcinomas showed homogeneous basolateral staining for VU-1D9 and strong apical staining for CEA; the moderately differentiated adenocarcinomas showed membranous as well as cytoplasmic VU-1D9 staining and luminal as well as cytoplasmic CEA staining. The VU-1D9 and CEA localizations and the stage of expression in relation to tumor progression were completely different. Strong CEA expression was seen in the adenomatous stage, while the homogeneous VU-1D9 expression required tumor progression to the carcinomatous stage. VU-1D9 especially when applied in combination with CEA, will be a useful marker for colorectal lesions, and its reactivity patterns in carcinoma can predict the prognosis. PMID- 9854476 TI - Classification of acute myeloid leukaemia in trephine biopsies with special reference to lactoferrin. AB - In spite of advances in immunohistochemical techniques, the histological subclassification of acute myeloid leukaemia (AML) in bone marrow biopsy has remained difficult. In particular, the translation of the diagnostic criteria of the French-American-British (FAB) cooperative group as primarily defined by bone marrow cytology into histology poses considerable problems. In this study, we investigated the expression of lactoferrin (LF) in various subtypes of AML and studied the usefulness of its immunohistochemical detection combined with a panel of antibodies directed against myeloperoxidase (MPOX), lysozyme (LYS), CD34 and naphthol-AS-D-chloroacetate esterase (NACE) staining in solving this problem. Trephine biopsies of 52 cases of AML were selected for histological evaluation in comparison to bone marrow aspirates classified according to FAB (M1 n = 10, M2 n = 7, M3 n = 11, M4 n = 13 and M5 n = 11). The results obtained confirmed the specificity of LF as a marker for secondary granules in neutrophilic myeloid cells and as a tool to subclassify AML. Its parallel application with (immuno )staining for MPOX, LYS and NACE has allowed the identification of M1, M3, M5 cases, where there LF is lacking. Typically M2 is characterized by a subpopulation of LF-positive cells which tend to display a myelocytic differentiation. However, M4 shows a heterogeneous expression pattern of LF: M4a may be defined as more immature variant without LF expression while in M4b a more mature myeloid subpopulation stains positive for LF. PMID- 9854477 TI - The prognostic significance of immunohistochemically detected p53 protein expression in non-small cell lung cancer treated with radiation therapy. AB - The prognostic significance of nuclear p53 protein expression in survival and local control was investigated immunohistochemically in 36 patients with inoperable or unresectable non-small cell lung cancer who were treated with radiation therapy (RT). Formalin-fixed, paraffin-embedded sections obtained by bronchoscopy were used to examine the expression of nuclear p53 protein with immunohistochemistry. In 25 cases (69%), p53 protein expression was detected. There was no relation between p53 expression and other pretreatment characteristics. Response to RT was found in all p-53 negative cases versus 72% in p53-positive cases (p < 0.05). The 2-year survival rate for p53-negative cases was 51% with a median survival time of 21 months. The corresponding rate for p53 positive cases was 31% with a median survival time of 9 months. This difference, however, did not reach a statistically significant level because of the small sample size. In conclusion, the results of this study suggest that p53 protein expression may be of predictive value on response to RT in non-small cell lung cancer. PMID- 9854478 TI - Bcl-2 and p53 expression in stage I endometrial carcinoma. AB - The role of p53 and bcl-2 apoptosis related proteins in the pathogenesis of endometrial cancer remains unclear. We immunohistochemically examined 133 surgically removed tumour specimens from patients with stage I endometrial cancer for p53 oncoprotein nuclear expression and bcl-2 cytoplasmic reactivity. 114/133 (86%) cases were of the endometrioid histological sub-type. A cut-off point of 10% of cells with strong reactivity was used for positivity. Positive p53 expression was observed in 12/133 (9%) and positive bcl-2 in 40/133 (30%) cases. p53 expression was not related to bcl-2 expression. No association of p53 and bcl 2 with depth of myometrial invasion, vascular invasion or histological grade was observed. However, continuous variable analysis revealed a trend of low grade cases to have a higher percentage of bcl-2 positive cells (16.3 + 27% vs. 7.8 + 16%; p = 0.09, unpaired two tailed t-test). Interestingly, a statistically significant association of p53 positivity with age was also observed (p = 0.03). A strong association of high grade with depth of myometrial invasion (p = 0.006) and vascular invasion (p = 0.0001) was also noticed. In addition, the presence of adenomyosis was also associated with low grade (p = 0.01) and absence of vascular invasion (p = 0.02). We conclude that although loss of bcl-2 protein expression and p53 mutant protein nuclear accumulation are early events in the endometrial cancer progression, histological grade and vascular invasion remain the most important factors defining local invasive behaviour of endometrial cancer. PMID- 9854479 TI - C-erbB2 expression predicts response to preoperative chemotherapy for locally advanced breast cancer. AB - BACKGROUND: Previous work suggests that the presence of c-erbB2 oncoprotein immunostaining and the proliferation rate of tumours, may be relevant to chemo sensitivity in breast cancer. PATIENTS AND METHODS: To investigate this we assessed pretreatment biopsies from 50 patients with locally advanced breast cancer for expression of c-erbB2 and MIB1 (proliferative marker) in relation to clinical response after 3 months preoperative chemotherapy. RESULTS: Objective response was significantly more likely (22/30, 73%) for tumours negative for c erbB2 membrane staining, compared to positively staining tumours (6/20, 30%, p = 0.0025). The percentage of cells staining positively for MIB1 was not predictive of response (p = 0.56). CONCLUSIONS: This study has shown an increased likelihood of response to preoperative chemotherapy for breast cancers negative for c-erbB2 staining. Previous studies have shown that c-erbB2 immunostaining can correlate with either chemo-resistance or chemo-response. We postulate that this conflict may be due to differences in the type of chemotherapy administered. This raises the possibility of biological markers being use to assist in the selection of the type of chemotherapy regimen administered to particular tumour biological phenotype. PMID- 9854480 TI - rIL-2 in metastatic soft tissue sarcomas refractory to chemotherapy: response and enhancement of further chemosensitivity. AB - Chemotherapy (CT) in advanced and metastatic soft tissue sarcoma (STS) has a reported response rate of 17 to 30%, with a median survival of 30 months. Experimental and clinical studies have demonstrated some response with high-dose intravenous recombinant Interleukin-2 (r-IL2). We report a retrospective analysis of twelve patients with metastatic or locally evolutive STS who received high dose rIL-2 immunotherapy: all patients were pre-treated with CT and were non responsive. All of these patients developed usual severe toxicity. One patient achieved a partial response with rIL-2 with more than 75% regression of lung metastases. All patients received CT post rIL-2 and 2 of them achieved a further response: 1 of them had a complete response (CR), and one had a partial response (PR). These results suggest that rIL-2 has a relatively low efficacy in patients with STS but that some of these patients can be, after rIL-2, responders to CT despite previous resistance. PMID- 9854481 TI - Forty years of monitoring head and neck cancer in Norway--no good news. AB - BACKGROUND: Male incidence rates of head and neck cancer are rising in most regions of the world. This paper describes Norwegian time trends in male incidence and relative survival from cancer of major head and neck sites. MATERIAL AND METHODS: The presentation comprises the entire Norwegian male population from 1955 to 1994. Incidence rates and relative survival rates were calculated for cancer of oral sites, oro-/hypopharynx and larynx separately. RESULTS: Age-adjusted incidence rates of oral and pharyngeal cancer increased after 1970 by 11 percent per 5-year period (95% CI: 5-19) and 14 percent per 5 year period (95% CI: 4-24), respectively. Laryngeal cancer incidence increased after 1955 by 14 percent per 5-year period (95% CI: 7-21). There were only small improvements in 5-year relative survival rates from oral and pharyngeal cancer. CONCLUSION: Male incidence rates increased for all the studied sites during the observation period. The prognosis for the patient groups included in this presentation has not improved substantially since the 1950's. PMID- 9854482 TI - Inhibition of melanogenesis as an adjuvant strategy in the treatment of melanotic melanomas: selective review and hypothesis. AB - Melanogenesis generates immunosuppressive and mutagenic environment and alters cellular metabolism. Melanin can protect malignant melanocytes against chemo-, radio-, and photodynamic therapy. Therefore, we propose to inhibit melanogenesis as an adjuvant treatment modality during immuno-, radio-, chemo- and photodynamic therapy, and as a measure of reducing the probability of melanoma progression. PMID- 9854483 TI - Prognostic index models in stage I and II endometrial carcinoma. AB - BACKGROUND: In most studies, authors recommend the use of independent variables in clinical decision-making, but no guidelines are given about how to use all the extracted information. MATERIALS AND METHODS: By combining two or three prognostically independent variables and using their relative prognostic impact (results from Cox analyses), a novel way of identifying high- and low-risk groups was developed. A total of 336 women with stage I-II endometrial carcinoma of medium or high risk were included. Twenty-one clinico- pathological variables, were initially studied in univariate analyses and significant variables were used for the construction of prognostic indices. Six prognostic indices were constructed, which then were used to calculate individual index values for each patient. RESULTS: The index values were highly prognostic and used for the identification of a limited high-risk group (10% of the patients) and a large low risk group (90% of the patients). Indices 1-3 were constructed for pre-treatment situations and included age, degree of differentiation, nuclear grade and S-phase fraction in various combinations. They identified a high- and a low-risk group with a 5-year disease-specific survival of 31-37% and 74.79%, respectively. Indices 4 and 5 were constructed using variables available after treatment and index 6 for patients with no evidence of disease (NED) after treatment completion. The 5-year survival for the latter indices were 30-52% and 75-85%, respectively. CONCLUSION: The use of prognostic indices in different clinical situations was generally superior to the use of single variables for the identification of well-defined high- and low-risk groups. PMID- 9854484 TI - Vascular endothelial growth factor in angiosarcoma. AB - Angiosarcoma(AS) is a malignant tumour of the endothelium of lymphatic or blood vessels. Vascular endothelial growth factor (VEGF), a specific mitogen of endothelial cells, is considered essential for the growth of many tumours including AS. We have recently treated several patients with AS; in two of them, using a newly developed enzyme-linked immunosorbent assay (ELISA) for VEGF, we sequentially measured serum VEGF concentrations. While serial serum VEGF levels reflected tumour burden, the various VEGF concentrations were within the normal range we reported previously. Furthermore, the VEGF concentrations were not remarkably elevated in AS tumour tissue compared with benign vascular lesions and hypervascular tumours. PMID- 9854485 TI - Radiation therapy and concurrent daily intra-arterial infusion of low-dose cisplatin following alteration of pelvic blood flow for invasive bladder cancer: a preliminary report. AB - Concurrent intra-arterial infusion chemotherapy and radiation therapy following alteration of pelvic blood flow was performed in 15 patients with invasive bladder cancer (T2: 2, T3a: 4, T3b: 5, T4: 4). Infusion chemotherapy consisted of a daily low-dose of cisplatin with a dose range of 7 to 9 mg per patient. Of the 15 patients, 11 achieved complete response (73%) and other had partial response. Four of those with CR developed recurrence (local recurrence in three and distant metastasis in one patient). Cause specific and disease-free survival at 3-years were 56% and 49.9%, respectively, and the bladder preservation rate at 3-year was 47%. Toxic reactions related to the treatment was of a reasonable level, and cisplatin-induced renal dysfunction was not experienced, even in the patients with poor renal function, although transient neutropenia and thrombocytopenia occurred in all patients at the latter stage of the treatment. This treatment modality is considered to be effective and feasible even in patients with locally advanced disease and/or those unsuited for cisplatin-based systemic chemotherapy. PMID- 9854486 TI - Ectopic breast cancer: a case report and review of the Japanese literature. AB - We report a rare case of ectopic breast cancer in a 68-year-old woman, successfully treated by surgery and chemotherapy. We found 37 cases including our case in the Japanese literature between 1933 to 1997. Our review demonstrated that the clinical features of Japanese patients who developed ectopic breast cancer was similar to previous foreign reports. PMID- 9854487 TI - Fine needle aspiration biopsy in the preoperative management of patients with thyroid nodules. AB - The aim of this study was to evaluate the role of Fine Needle Aspiration Biopsy (FNAB) in the preoperative diagnostic management of patients with solitary or dominant thyroid nodules. This study was performed on 1054 patients followed for various thyroid disorders during a three year period (1992-1995). One hundred thirty eight patients were surgically treated, 61 after cytological indication and 77 after clinical indication, of these, 67 were cytologically negative and 10 were not conclusive. The cytological diagnosis was compared to the final histological result. Among the 138 cases, 27 were malignant, 39 were adenomas and 72 were benign lesions. Of the 67 cytologically negative cases, 63 (94%) were histologically benign, three were papillary carcinomas and one was an oxyphilic adenoma. Of the 45 cytologically suspicious aspirates, four were malignant neoplasias, 38 were follicular adenomas, two were hyperplastic goiters and one was an Hashimoto thyroiditis. All the 16 cytologically positive cases, were confirmed histologically. The limits of FNAB, which emerge from our and other studies, were based mainly in the difficulty of discriminating follicular adenomas from well differentiated follicular carcinomas. Nevertheless, FNAB with ultrasonographic support, has been unanimously accepted as a guide test in the selection of patients with thyroidal pathology who need surgery. PMID- 9854489 TI - Biomagnetic activity in ovarian lesions. AB - BACKGROUND-MATERIALS: This study aimed to investigate biomagnetic activity in benign and malignant ovarian diseases using the biomagnetometer SQUID. Magnetic recordings were obtained from 40 patients with palpable ovarian lesions. 19 of these were invasive carcinomas, and 21 were benign ovarian lesions. METHODS: We used a one channel biomagnetometer SQUID (superconducting quantum interference device), in order to measure the magnetic field from benign and malignant ovarian diseases. RESULTS: Interestingly, the ovarian lesion waveforms and the corresponding spectral densities were of high amplitude in most (96%) malignant ovarian lesions, and of low amplitude in most (95%) benign ovarian diseases. These findings were of statistical significance (students t-test p < 0.005). CONCLUSIONS: It is suggested that biomagnetic measurement of benign and malignant ovarian diseases, which is an entirely new application of SQUID technology, is a promising procedure for assessing ovarian tumors. PMID- 9854488 TI - Evaluation of circulating tumour cells expressing CD44 variants in the blood of gastric cancer patients by flow cytometry. AB - The occurrence of circulating tumour cells in the blood of 51 patients with gastric cancer (stages I-IV) was studied using flow cytometry, cell sorting, immunohistochemistry and reverse transcription polymerase chain reaction (RT PCR). The lysed whole blood samples were stained with monoclonal antibodies against common leukocyte antigen (CD45), epithelial membrane antigen (EMA), tumour associated glycoprotein 72kD (TAG72), CD44 variants (v5 and v6) and analysed by flow cytometry within ungated or CD45-gated populations. The frequency of detection of TAG72+, CD44v5+ and v6+ cells within CD45- gate was considerably increased in comparison to the ungated population. Furthermore, the presence of tumour cells was directly demonstrated by immunostaining for cytokeratin 18 of sorted CD45- population. The presence of CD44v5+, v6- cells and CD44v-mRNA in the blood was compared to their expression in the primary tumour. The occurrence of circulating CD44v+ cells was associated with their presence in the primary tumour and CD44v-mRNA in the blood. The method described may provide a sensitive tumour marker-independent tool for detection of circulating tumour cells in cancer patients. PMID- 9854490 TI - P53 mutation in advanced stage of primary myelodysplastic syndrome. AB - Polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) analysis of the p53 tumor suppressor gene (from exon 2 to 9) was performed on samples from 47 adult patients with primary myelodysplastic syndrome (MDS). Point mutation was found in 5 (11%) patients: exon 7 in 3, exon 4 in 1 and intron 5 in 1. The frequency of p53 mutation was significantly higher at advanced stages (p = 0.048) and higher in patients with abnormal karyotypes (p = 0.023). Although all of the p53 mutations were detected at advanced stages, four of them were detected at initial diagnosis with very short survival. Sequential SSCP analysis in 20 transformed MDS patients revealed only one new p53 mutation during progression from early MDS phases. The results suggest that p53 mutation might occur as an early genetic event and is probably associated with rapid progression and poor survival in some MDS patients. PMID- 9854491 TI - Serum anti-p53 antibodies in the follow-up of patients with advanced ovarian carcinoma. AB - Serum anti-p53 antibodies have been detected in different human malignancies, including ovarian carcinoma. In the present investigation these autoantibodies were retrospectively measured with a new ELISA (Immunotech, Marseilles, France) before first surgery and subsequently at different times during the course of disease from 40 patients with advanced ovarian carcinoma. Anti-p53 antibodies were preoperatively found in 15 (37.5%) patients. With regard to the follow-up of these 15 patients, anti-p53 antibodies were detected in 87.8% of the 41 samples drawn when there was clinical evidence of disease compared to 57.1% of the 14 samples collected when there was no clinical evidence of tumor (p = 0.037). As for the 25 patients whose serum originally scored negative, the autoantibodies were found only in 1.8% of the 113 samples obtained during the follow-up, independently of the status of disease. In conclusion, anti-p53 antibodies are often detected in serum from patients with advanced ovarian carcinoma. However, the serial measurement of these autoantibodies does not seem to give useful clinical information for the follow-up of these patients. PMID- 9854492 TI - Comparison of microvessel density with tumor associated macrophages in invasive breast carcinoma. AB - Neovascularization, the growth and formation of capillary blood vessels, is an essential component of solid tumor growth and a critical step in metastasis. Tumor associated macrophages (TAMs) have several functions related to tumor biology including growth, proliferative rate, stroma formation and dissolution, and neovascularization. The aim of this study was to define the TAM and microvessel density (MD) in human invasive breast carcinoma NOS and to correlate their values with lymph node status, tumor size, tumor grade and mitotic activity index (MAI), and, finally, to determine whether MD is connected with TAMs. A total number of 57 invasive breast carcinomas NOS were processed for immunohistochemical analysis using mAb to F-VIII to visualize endothelial cells and mAb to CD68 antigens for macrophages. Statistical analysis showed only a positive correlation between TAMs and MAI (p = 0.004). These results support the notion that intensity of tumor angiogenesis does not provide additional prognostic significance, while TAMs may play a positive role in breast cancer micro system since they regulate tumor proliferation. PMID- 9854493 TI - Limitations in the use of glutathione S-transferase P1 in urine as a marker for bladder cancer. AB - BACKGROUND: GST pi (GSTPl) is overexpressed in bladder cancer and desquamation of the tumour may produce detectable levels of urinary GSTPl which could be used as a marker for the early diagnosis of bladder cancer. MATERIALS AND METHODS: A preliminary study in 27 transitional cell carcinoma (TCC) patients and 20 controls, using an ELISA methodl is presented here. RESULTS: 55.5% of TCC patients were positive for GSTP1, while all control samples were negative. Some of the GSTP1 positive cases also gave positive results for haematuria, which indicates that a limitation of this marker involves the contamination of the urine with erythocyte GSTP1. In 5 cases (18.5%) without haematuria detectable levels of GST pi were found. CONCLUSIONS: Further studies would be required to assess the advantages of this technique over clas sical cytology or as a complement to it, especially in patients in a phase of temporary negative haematuria. PMID- 9854494 TI - p53 expression affects the efficacy of adjuvant chemotherapy after resection of invasive ductal carcinoma of the pancreas. AB - p53 tumor suppressor gene has a dual role as a trigger of apoptosis and as an initiator of DNA repair, suggesting its involvement in the mechanisms of drug resistance or chemosensitivity. The present study assessed the implication of p53 expression in the prognosis of patients and the efficacy of adjuvant chemotherapy for resectable invasive ductal carcinoma (IDC) of the pancreas. A total of 58 patients with primary IDC of the pancreas underwent pancreatectomy between 1982 and 1996: 28 patients received surgery alone and 30 patients received postsurgical adjuvant chemotherapy. p53 protein was stained immunohistochemically with anti-p53 monoclonal antibody. p53 was positively expressed in 29 out of 58 primary lesions (50%), and the survival curve of the patients with p53 (+) pancreatic cancer is lower than that of those with p53 (-) cancer. On the other hand, the survival curve of adjuvant chemotherapy group was also higher than that of surgery alone group, and furthermore, in patients with p53 (+) cancer, the survival curve of adjuvant chemotherapy group was significantly better than that of the surgery alone group. A multivariate analysis showed that p53 expression or adjuvant chemotherapy is not a significant risk-factor for prognosis, but that adjuvant chemotherapy is a significant risk factor for the patients with p53 (+) pancreatic cancer, which suggests that p53 expression affects the efficacy of chemotherapy. p53 expression may be beneficial as an indicator for introduction of adjuvant chemotherapy in pancreatic cancer patients. PMID- 9854495 TI - Gastric Malt lymphoma: a clinicopathological study. AB - The detection of HP infection by means of non invasive methods such as breath test and urease test, which are adopted alternatively to histological examination, can delay the diagnosis of early lymphomas. On the other hand the proved regression of gastric low grade Malt lymphomas after a successful HP eradication strongly suggests the need for early diagnosis. The aim of this study was to report the results of our experience with regard to clinical, endoscopic, histopathologic, and immunohistochemical features of gastric MALT lymphomas. We studied twenty-seven consecutive cases of gastric Malt lymphoma. HP presence in gastric mucosa was detected in all cases but endoscopy only in 40.7% of cases giving a diagnosis of suspected malignancy. In conclusion, it is very important to sample gastric mucosa in HP positive patients because the histological examination represents the most effective tool to detect lesions at the earliest and curable stage. PMID- 9854496 TI - Quantification of cathepsin D in plasma of patients with malignant melanoma. AB - The high mortality rate of melanoma patients who develop metastases prompted us to seek for a prognostic soluble marker to identify high-risk and non-risk patients at the stage of the primary tumour. Therefore, we developed a new ELISA for quantifying plasma concentrations of the proteolytic enzyme cathepsin D (CD) in patients with primary tumours (MM-P) and with metastases (MM-M), respectively, compared to a control group. Whereas healthy probands (n = 56) and MM-P (n = 68) showed similar mean values of CD (0.73 +/- 0.45 ng/ml and 0.82 + 0.80 ng/ml), MM M (n = 40) yielded significantly reduced plasma levels (0.43 +/- 0.53 ng/ml) revealing a high significant discrimination both between controls and MM-M, and MM-P and MM-M (p < 0.0001). From the beginning of the study (1990) to the present 11 of 68 MM-P developed metastases. In order to test the prognostic efficiency of this enzyme to determine those patients at high-risk and non-risk for developing metastases, the receiver operating characteristic analysis was used showing that CD plasma levels cannot supply reliable prognostic values (W = 0.53, p = 0.66). PMID- 9854497 TI - Soluble HLA class I antigens in plasma of patients with malignant melanoma. AB - The high mortality rate of melanoma patients who develop metastases prompted us to look for prognostic markers to determine high-risk and non-risk patients at the primary tumour stage. Therefore, we quantified plasma concentrations of soluble HLA class I antigens (sHLA-I) by ELISA in patients with primary tumours (MM-P) and with metastases (MM-M), respectively, and compared them to a control group. Whereas healthy probands (n = 55) and MM-M (n = 38) showed similar mean values of sHLA-I (1.30 +/- 1.44 micrograms/ml and 1.29 +/- 1.27 micrograms/ml), MM-P (n = 67) revealed significantly reduced levels of this marker (0.84 +/- 0.85 microgram/ml). This result matches with our immunohistological staining of membrane-bound HLA-I in sections of paraffin-embedded melanoma. Further subdivision of the MM-P substantiated the observation that mean values of decreased sHLA-I concentrations are in line with high tumour thickness. Since the beginning of this study (1990) to date, 11 of 67 MM-P have developed metastases. The prognostic efficiency of sHLA-I to identify high-risk and non-risk patients was tested by ROC-analysis (receiver operating characteristic) and did not demonstrate good prognostic relevance for sHLA-I (W = 0.64, p = 0.04). PMID- 9854498 TI - Steroid receptor concentrations as a prognostic factor in atypical endometrial hyperplasia. AB - Patients with atypical adenomatous hyperplasia run the risk of developing endometrial carcinoma in about 30% of cases, but about 60% normalise without any treatment. High-dose gestagen treatment cures almost 100% but as only 40% need treatment a prognostic factor to identify those at risk would be useful. Using material from 52 patients we investigated whether steroid receptor concentrations could be that factor. However, we could not identify any association between histopathology after a short follow-up and steroid receptor concentrations. PMID- 9854499 TI - Neuro- and ototoxicity of high-dose carboplatin treatment in poor prognosis ovarian cancer patients. AB - BACKGROUND: Hematopoietic toxicity of high-dose carboplatin (HD-CBDCA) chemotherapy can be managed effectively with autologous blood cell support, but no conclusive data are available on its neuro- and ototoxicity. PATIENTS AND METHODS: We determined the neuro- and ototoxicity of HD-CBDCA in 10 patients affected by advanced ovarian cancer. HD-CBDCA was delivered as 24-hour continuous infusion or as 5-day schedules. Each patient underwent an extended clinical and instrumental neurological and otological evaluation before, during and after treatment. RESULTS: After HD-CBDCA only 1 patient had a clinically-evident peripheral neuropathy, while 3 additional patients had only distal paresthesias. Neurophysiological examination evidenced mild, although diffuse, sensory nerve impairment. Motor nerve impairment was also occasionally observed. All the sensory and motor pathological changes had a favorable course during the follow up period. Ototoxicity was more severe than neurotoxicity and, in one case it was dose-limiting and audiologic impairment tended to remain constant also in the follow-up period. CONCLUSIONS: HD-CBDCA treatment can be tolerated by most of the patients, but careful monitoring of neuro- and, especially, ototoxicity should be planned. PMID- 9854500 TI - Comparison of nuclear matrix protein 22 and bladder tumor antigen in urine of patients with bladder cancer. AB - Recently, two new tumor markers for bladder cancer have been introduced: NMP22 test and BTA TRAK assay. This study was designed to evaluate the urinary values of these two proteins using quantitative enzyme immunoassays in well microplates. Urine samples from 47 healthy subjects, 26 with benign genitourinary disorders and 109 patients with a histological diagnosis of bladder cancer were collected. The specificity, the positive predictive value, the negative predictive value and the efficiency were established for NMP 22 and BTA, and the cut off values were fixed at a specificity of 95% in the benign disease group (12 U/ml and 23 U/ml respectively). We observed a very high concordance between the two urinary tumor markers (73%), although the overall sensitivity of BTA in bladder cancer patients seems to be better than that of NMP22 (62% vs 54% respectively), especially in the superficial disease group (36% for BTA and 14% for NMP22). PMID- 9854501 TI - Combined chemoradiotherapy for invasive bladder cancer: a feasibility approach in a non selected population. AB - BACKGROUND: We aimed to evaluate the tolerance and response rate to neo-adjuvant combined chemo-radiotherapy and to determine the possibilities of conservative treatment in patients (pts) with muscle invasive bladder cancer. METHODS: Thirty nine T1-4, NO-3 pts were enrolled in this study. After 2 cycles of treatment, responders at intermediate assessment were treated by either additional chemotherapy +/- radiotherapy or radical cystectomy. If there was evidence of persistent tumour, radical cystectomy was performed whenever possible. RESULTS: Thirty-five (90%) pts completed the pre-operative treatment, 26/39 (67%) were in remission at intermediate assessment and 8 of them underwent a radical cystectomy. The median follow-up was 87 months, and 15 pts are alive without evidence of tumour and 5 with bladder preservation. The 3 and 5-year survival was 51 and 40%, respectively. Median survival differences between the responding operated or non-operated pts were not statistically different. CONCLUSIONS: Neo adjuvant chemo-radiotherapy for invasive bladder cancer is effective with acceptable toxicity, and inoperable patients can benefit from such treatment. PMID- 9854502 TI - TNF alpha in isolated perfusion systems: success in the limb, developments for the liver credits, debits and future perspectives. AB - The clinical applicability of Tumor Necrosis Factor-a pi (TNF) is under renewed investigation because of its successful use in the isolated limb perfusion in patients with irresectable soft tissue extremity sarcomas. The high response rate of > 80% with a similarly successful limb salvage rate in this patient population has led to the submission of TNF for registration for this indication in Europe. Similarly, the agent has been shown to be successful in the isolated limb perfusion setting for tumors other than sarcomas, e.g. melanoma, carcinomas. This has caused renewed interest in TNF alpha and in its application in isolated organ perfusions, such as isolated hepatic perfusion. At the Rotterdam Cancer Center a preclinical-clinical interactive development program has been established dedicated to isolated limb, kidney, liver and lung perfusions and the application of new drugs such as TNF and TNF-mutants in these systems. Moreover a program dedicated to reduce the magnitude of surgical procedures by using occlusion balloon catheters is investigating the techniques and pharmacokinetics of procedures such as balloon catheter mediated hypoxic pelvic perfusions (HPP) and isolated hypoxic hepatic perfusions (IHHP). Here we present an overview of these developments. PMID- 9854503 TI - The search for a potent DNA vaccine against AIDS: the enhancement of immunogenicity by chemical and genetic adjuvants. AB - DNA vaccination is a new way to generate antigen specific immune responses by inoculating plasmid DNA encoding microbial genes. We believe that the use of immunologic adjuvants together with DNA vaccines is a promising way to enhance and optimize DNA-derived immunity. Various chemical and genetic adjuvants have been explored in our recent study. Data from these studies suggests that adjuvants exert their immunomodulatory properties through several mechanisms such as lymphoid cells recruitment, cytokine induction, and the facilitation of DNA entry into cells. Furthermore, each adjuvant has unique immunomodulatory effects on cell-mediated and humoral immune responses induced by DNA vaccination to human immunodeficiency virus type 1. To optimize the desirable immune response, the choice of adjuvant and decision of immunization route are important considerations. A similar approach may be useful potentiating anticancer immunotherapy as well. PMID- 9854504 TI - The past, the present and future of the OK-432 therapy for patients with malignant effusions. AB - For the past 20 years, our group has treated over 400 cases of malignant effusion by the intraperitoneal injection of streptococcal preparation OK432 (OK-432 therapy) and has investigated extensively the antitumor mechanisms of this therapy. Prospective clinical data has demonstrated that the OK-432 therapy induced a definite reduction of the effusions in around 60% (responders) of cases and significantly prolonged the survival time in patients who responded well. In addition, a definite reduction of original tumor mass volume was found in around 20% of cases. We have shown that OK432-induced neutrophils, lymphocytes, and probably macrophages may play an important role in tumor cell destruction in ascites. Tumor necrosis factor alpha (TNF-alpha)-induced CD11b/CD18 expression on leukocytes and interferon-gamma (IFN-gamma)-induced ICAM-1 expression on tumor cells may play an important role in leukocyte-mediated tumor destruction. It has also been shown that OK-432 induces various cytokines, such as TNF-alpha, TNF beta, IFN-alpha IFN-gamma, interleukin-1 (IL-1), IL-2, IL-6, IL-12, tumor growth inhibitory factor(s) (TGIF), and possibly unknown apoptosis-inducing factor(s). Some of these cytokines have been adduced as representing the antitumor activity. These data suggest that two pathways of antitumor activity, i.e., cell-mediated and cytokine-mediated, can be induced simultaneously in the peritoneal cavity. OK 432 therapy may be valuable in the management of patients with malignant effusions. Future clinical and basic research should contribute to further progress in OK-432 therapy. PMID- 9854505 TI - The possible role of sense and antisense peptide interactions in the generation and maintenance of the tertiary structure of a protein. AB - Antisense peptide is an amino acid sequence translated from antisense sequence of mRNA for a peptide sequence (sense peptide). Previous evidence indicates that sense and antisense peptides have a tendency to interact with each other. We expected that this kind of interaction might be involved in the formation and maintenance of tertiary structure of protein molecules. Many amino acids have several RNA codons and we regarded amino acids coded by anti-codons of all of codons of each amino acid as antisense amino acids. On this stand point, we generated a computer program to search for antisense peptides. Then we found that there are peptide sequences which are antisense to several peptide sequences in a protein molecule, and these peptide sequences have been termed as antisense homology boxes (AHB). AHB peptides were synthesized and some of them had a potent capacity to interfere with the function of the molecule. PMID- 9854506 TI - Endogenous TNF induction therapy using rTNF-SAM2 in patients with pulmonary metastases from colorectal cancer. AB - BACKGROUND: Distant metastases from colorectal cancer are generally refractory to conventional therapies, with the exception of surgical resection. The aim of this study was to evaluate the clinical application of endogenous TNF induction therapy by using a mutant TNF (rTNF-SAM2) as a primer in endogenous/exogenous TNF therapy (EET therapy) in patients with pulmonary metastases from colorectal cancer. METHODS: The subjects were 17 patients, 5 of whom underwent EET therapy alone and 12 of whom underwent EET therapy and the administration of anticancer agents. RESULTS: Partial response was observed in 6 patients (50%) who underwent EET therapy with anticancer agents. In seven (53.8%) of 13 patients who showed a high serum CEA value, their CEA levels were considered to be improved. Severe toxic effects occurred in 3 of the patients studied (17.6%). The mean survival was 26.0 months among those with a partial response and 16.6 months among those with no change. No significant difference was observed between these two groups. Histological assessments indicated that tumor necrosis, fibrosis and cellular infiltration tended to intensify in cases treated with EET therapy compared with the cases who received surgery alone. CONCLUSION: EET therapy with anticancer agents is well-tolerated and effective for pulmonary metastases from colorectal cancer. PMID- 9854507 TI - Clinical effects of exogenous/endogenous TNF therapy on metastatic lesions of 34 colorectal cancer patients. AB - BACKGROUND: We performed endogenous/exogenous TNF (EET) therapy using, as a primer, recombinant TNF-SAM2 (rTNF-S) as a premier; it has a broader anti-cancer spectrum and is less toxic than conventional TNF, and OK-432 as a trigger in patients with metastasis of colorectal cancer to the lung and/or liver. METHODS: The subjects consisted of 34 patients of whom 8 had lung metastasis, 17 liver metastasis, and 9 lung and liver metastasis. EET therapy was performed without chemotherapy in 11 patients and with anti-cancer drugs such as mitomycin-C (MMC) and 5-FU in the other 23. RESULTS: No patients showed a complete response. Partial response was observed in 10 patients (29.4%), minor response in 8 (23.5%), no change in 14 (41.2%), and progressive disease in 2 (5.8%). The response rate was 29.4%. A greater effect was observed in patients treated with EET in combination with anti-cancer drugs than those treated with EET alone. No serious side effects were observed, although all patients developed a fever above 38 degrees C, chill, and shiver. CONCLUSIONS: EET therapy with rTNF-S combined with anti cancer drugs chemotherapy may be effective, and further studies are needed to select the most suitable anticancer drugs for combination with, and to determine the effective frequency of treatment. PMID- 9854508 TI - Clinical experience of EET therapy for 75 advanced cancer patients. AB - BACKGROUND: Exogenous/endogenous TNF (EET) therapy has a strong effect which causes inflammation in the body. Together with the antitumor effect of a single administration, the complementary effect of immunotherapy based on specific immunostimulation of tumor cells is also expected. We studied the characteristics of EET therapy in cases of advanced cancer. PATIENTS AND METHODS: The patients were 75 cases with advanced cancer including 48 cases of colon cancer, 10 cases of urological cancer, and 9 cases of gynecological cancer. One course of therapy was composed of the intravenous administration of either IFN-gamma or TNF-SAM2 followed 3 hours later by OK-432. At least 2 courses were repeated during a 2 weeks period. RESULTS: As response cases, partial response (PR) was observed in 7 of the 42 cases of colon cancer (17%) which had an evaluable lesion. Objective responses of lung metastases were found in the patients with multiple organ metastases of cervical cancer, ovarian cancer and uterine rhabdomyosarcoma. The group of colorectal cancer patients with liver metastases which underwent more than 5 courses of the therapy showed a longer survival period compared to the control group. One of the side effects, transitory hypotension, was observed in 46% of the cases. CONCLUSION: In spite of restricted objective responses to date with EET therapy alone, anticancer effects observed in various kinds of tumors and activation of a cytokine network by which Th1 cells might be specifically activated suggest that a new biotherapy based on EET therapy may have potential. PMID- 9854509 TI - Impaired production of Th1 cytokines and increased frequency of Th2 subsets in PBMC from advanced cancer patients. AB - BACKGROUND: A reported imbalance of Th1 and Th2 may be responsible for the occurrence and progress of certain diseases. Patients with advanced cancer may have impaired cell-mediated immunity caused by a switch from Th1 to Th2. We investigated the balance of Th1 and Th2 in cancer patients using two approaches. METHODS: Flow cytometry was used to define Th1 and Th2 using intracellular cytokines, thereby determining the frequency of these helper T cells in CD4+ peripheral blood mononuclear cells (PBMC). The amount of cytokines secreted by stimulated PBMC in bulk culture was determined. RESULTS: Production of TNF-alpha and IFN-gamma in the culture was significantly lower in 9 patients (1188 +/- 634 pg/ml and 46 +/- 84 pg/ml) than in 10 healthy subjects (2491 +/- 1037 pg/ml and 295 +/- 219 pg/ml) (p < 0.002 and p < 0.003), while the production of IL-4 was slightly higher in cancer patients than in the healthy subjects. The percentage of IFN-gamma-IL-4+ cells in CD4+ cells was significantly higher in the patients than in healthy subjects (4.3 +/- 2.0% and 2.4 +/- 0.7%); there was no difference in IFN-gamma+IL-4- cells between the two groups. The ratio of IFN-gamma+IL-4- and IFN-gamma-IL-4+ cells per individual was significantly lower in the patients. CONCLUSION: These results indicated that an imbalance of Th1 and Th2 was found not only in the frequency of the subsets in PBMC, but also in the capacity for cytokine-production. PMID- 9854510 TI - Mechanisms by which chemotherapeutic agents augment the antitumor effects of tumor necrosis factor: involvement of the pattern shift of cytokines from Th2 to Th1 in tumor lesions. AB - BACKGROUND: The antitumor effect exerted by tumor necrosis factor (TNF) is characteristic in that it causes central necrosis of the tumor mass. Viable tumor cells surrounding the tumor mass remain, however, even after most of the mass is necrotized, and these cells gradually regrow and form tumors. To overcome this, we analyzed the combined effects of chemotherapeutic agents used with TNF. Alkylating agents such as cyclophosphamide altered the antitumor effect qualitatively, leading to complete regression which TNF alone could not achieve. The mechanism, behind the enhancement of endogenous TNF production and expression of mRNA of various cytokines by the combination of chemotherapeutic agents with TNF inducer was investigated in Meth A fibrosarcoma. METHODS: Seven days after the inoculation of Meth A fibrosarcoma into BALB/c mice, cyclophosphamide (CY, 100-150 mg/kg) was injected intraperitoneally, and 7 days later endogenous TNF was induced by the intradermal administration of lipopolysaccharide (LPS, 400 micrograms/kg) or intravenous injection of ONO-4007, a synthetic lipid A derivative (30 mg/kg). RESULTS: A combination therapy of LPS or ONO-4007 with CY showed the effect of complete regression in 50-100% of tested mice, while CY, LPSp or ONO-4007 alone did not cause complete regression. The amount of endogenous TNF induced by LPSp or ONO-4007 around a tumor lesion with CY was 4-5 fold higher than that without CY. The expression of mRNA of transforming growth factor-beta was suppressed by CY seven days after the injection, and expressions of mRNA of IL-1 beta and TNF-alpha were augmented by the administration of CY 1 to 3 hours after the administration of ONO-4007. CONCLUSION: Some chemotherapeutic agents thus appear to augment the antitumor effect of TNF around tumor lesions, leading to tumor regression through a mechanism in which the agent changes the host's immune status, especially around a tumor lesion and pattern shift of cytokines from Th2 to Th1. PMID- 9854511 TI - Treatment of malignant astrocytomas with recombinant mutant human tumor necrosis factor-alpha (TNF-SAM2). AB - We present our experience with a combination chemotherapy regimen consisting of ranimustine (MCNU) and recombinant human mutant tumor necrosis factor-alpha (TNF SAM2) for malignant astrocytomas. The initial regimens were prescribed as adjuvant therapy in conjunction with radiotherapy following standard surgical treatment. Newly diagnosed patients were treated with up to four cycles of this regimen (TNF-SAM2, MCNU, and radiotherapy: TMR group). Seventeen patients (11 men and 6 women) aged 24 to 68 years (median 54.6 years) were eligible and evaluated for response and toxicity. The estimated median survival time was 354 weeks with anaplastic astrocytomas, and 76 weeks with glioblastomas. One- and 2-year survival rates were 100% and 100% with anaplastic astrocytomas, and 69.2% and 29.7% with glioblastomas. Grade 3 and 4 hematological toxicities were not experienced. None of the patients experienced a treatment delay due to toxicity. All other acute toxicities were anticipated and manageable. Twenty three patients (11 men and 12 women) aged 22 to 66 years (median 50.7 years) were evaluated as a historical control of patients who received chemotherapy with MCNU alone in conjunction with radiotherapy following standard surgical treatment (MCNU and radiotherapy: MR group). The estimated median survival time was 205 weeks with anaplastic astrocytomas, and 62 weeks with glioblastomas. One- and 2-year survival rates were 88.9% and 66.7% with anaplastic astrocytomas, and 71.4% and 7.1% with glioblastomas in this group. There were no significant differences in survival rates between patients in the TMR and MR groups with either anaplastic astrocytoma or glioblastoma. However, despite the small number of patients, those with anaplastic astrocytoma in the TMR group tended to survive longer than those in the MR group. These results suggest that combined chemotherapy with mutant TNF alpha may benefit those with anaplastic astrocytoma, and thus warrants further evaluation. On the other hand, the lack of activity does not warrant any further study of this schedule of TNF-SAM2 for the treatment of glioblastoma. PMID- 9854512 TI - Worlds without borders. PMID- 9854513 TI - RE: JW Butany, TE David, M Ojha. Histological and morphometric analyses of early and late aortocoronary vein grafts and distal anastomoses. PMID- 9854514 TI - The genome--where to from here? PMID- 9854516 TI - Falling hospital mortality for acute myocardial infarction in Quebec hospitals. AB - OBJECTIVE: To present in-hospital mortality trends for acute myocardial infarction (AMI). DESIGN: Observational study using the Quebec administrative hospital database, which records all hospitalizations for AMI, for the period 1986 to 1996. RESULTS: From 1986 to 1996, the case fatality rate for AMI decreased from 18.4% to 12.7% despite an increase in the total number of admissions, due to an ageing population. Men and women have had similar yearly mortality reductions--7.6% versus 7.4%, respectively--although the absolute case fatality rate remains significantly higher for women. The mortality reduction for men was constant over the decade, while the decline for women was more pronounced over the last five years. Improving case fatality rates were also observed in the elderly and again were most evident from 1991 to 1996. CONCLUSIONS: These data show a sharp decline in case fatality rates for AMI patients treated in Quebec hospitals from 1986 to 1996, suggesting that treatment advances observed in clinical trials are being applied at a population level. While improved survival has been observed in all patient groups, the data suggest that the part of the decline in mortality may be due to increased penetration of proven treatment strategies in women and the elderly. PMID- 9854515 TI - The Study of Health Assessment and Risk in Ethnic groups (SHARE): rationale and design. The SHARE Investigators. AB - The Study of Health Assessment and Risk in Ethnic groups (SHARE) is a study to determine the risk factors for atherosclerosis among three ethnic populations in Canada. Three hundred and thirty South Asian Canadian, 320 Chinese Canadian and 320 European Canadian men and women between 35 and 75 years of age are being randomly sampled from communities in Hamilton and Toronto, Ontario and Edmonton, Alberta for assessment of conventional (i.e., smoking, dyslipidemia, diabetes and hypertension) and emerging (i.e., candidate genes for atherosclerosis, homocysteine, fibrinolytic parameters, neurohormones, glucose intolerance, markers of infection, socioeconomic status, psychosocial status and diet) cardiovascular disease risk factors. Subclinical atherosclerosis is measured by quantitative B-mode ultrasonography of the carotid arteries, and other objective measures of vascular disease are a 12-lead electrocardiogram, a two-dimensional echocardiogram, ankle to arm blood pressure ratio and urine microalbumin concentration. The relationship between the conventional and emerging risk factors, and atherosclerosis, vascular disease and markers of end-organ damage will be evaluated between and within ethnic groups. PMID- 9854517 TI - Rate control therapy for atrial fibrillation following coronary artery bypass surgery. AB - Atrial fibrillation (AF) is a common complication of coronary artery bypass surgery (CABS). Because conventional antiarrhythmic therapy may cause proarrhythmia, a rate control approach to AF was evaluated in 59 patients post CABS. The use of digoxin with or without verapamil for AF was associated with spontaneous conversion to normal sinus rhythm in 55 of 59 patients. Two to four weeks later, all patients were on digoxin and 12% were on verapamil, with two of four AF patients having converted to normal sinus rhythm and two others previously in normal sinus rhythm now in AF. A rate control approach to treating AF post-CABS resulted in over 90% of patients being in normal sinus rhythm for two to four weeks after the onset of the arrhythmia. PMID- 9854518 TI - Short-term recovery from cardiac surgery in women: suggestions for practice. AB - OBJECTIVE: To examine the effect of sex on short-term recovery from cardiac surgery. DESIGN: Prospective, nonrandomized design. Men and women undergoing elective cardiac surgery were interviewed in the immediate preoperative period and monthly by telephone through the third postoperative month. SETTING: Subjects were recruited from three cardiac surgery referral centres in western Canada. PATIENTS: One hundred and thirty-two consecutive patients entering the hospital for coronary artery bypass and/or valve surgery were recruited. Inclusion criteria required that subjects be able to communicate effectively and to live independently. One hundred and twenty subjects completed the study--60 men and 60 women; 30 of each sex group were less than 65 years of age and 30 were 65 years and older. MAIN RESULTS: Women were more functionally impaired, had less social support and reported lower life satisfaction than men preoperatively. There were no significant differences between the sexes in the outcome measures three months postoperatively, and neither age nor sex consistently predicted outcome at the three-month end-point. Sex-specific issues reflect the social circumstances from which women entered the process of having their heart disease diagnosed and undergoing cardiac surgery. CONCLUSIONS: Findings support recent conclusions that there are few differences between men and women in the recovery from cardiac surgery; however, the existing differences warrant serious consideration for changes in current clinical practice. Sex differences included women's poorer preoperative functional status, lesser social support and differences in the nature of work women return to following their surgery. PMID- 9854519 TI - Vascular and cardiac effects of amlodipine in acute heart failure in dogs. AB - BACKGROUND: Amlodipine improves exercise capacity in patients with chronic congestive heart failure (HF), but the mechanisms of this effect are unknown. OBJECTIVE: To test the hypothesis, in a canine model of acute, ischemic HF, that amlodipine increases vascular capacitance and reduces cardiac filling pressures. METHODS: Amlodipine was given to 13 anesthetized, splenectomized dogs (six controls and seven with HF). Aortic, left ventricular end-diastolic (LVEDP) and portal venous (Pportal) pressures, cardiac output, portal flow (ultrasonic probe) and intestinal blood volume (IBV, 99mTc blood-pool scintigraphy) were measured. Intestinal vascular conductance (= 1/resistance) and vascular capacitance (CAP) were measured before and 15 mins after repetitive 150 micrograms/kg dosages of amlodipine (maximum cumulative dosage, 1000 micrograms/kg). Pportal-IBV curves were obtained by impeding portal flow (pneumatic cuff), and change in CAP was defined by the change in IBV at Pportal = 7.5 mmHg. HF was induced by microsphere embolization of the left coronary artery. RESULTS: CAP increased in the control group (+ 28%, P < 0.01) but decreased (-9%, P < 0.05) in the HF group. Left ventricular stroke work increased in the control group (P < 0.05), while it decreased (P < 0.05) in the HF group, suggesting a negative inotropic effect. In the control group, LVEDP increased after amlodipine was given (P < 0.05) but did not change significantly in the HF group. CONCLUSIONS: In the acute experimental HF model, amlodipine failed to increase intestinal vascular CAP or decrease filling pressures, and may have had a negative inotropic effect. The experiment failed to demonstrate a beneficial hemodynamic effect of amlodipine in acute HF, and the mechanism of benefit of this agent in chronic HF remains unclear. PMID- 9854520 TI - Is brain cancer an occupational disease of cardiologists? AB - OBJECTIVE: To assess the plausibility of radiation as a cause of the statistically unusual event of two cardiologists in Toronto, Ontario who were diagnosed with brain tumours in 1997. DESIGN: Computation of the expected occurrence of brain cancer in Ontario cardiologists and review of the epidemiological literature pertaining to radiation and brain cancer. DATA SYNTHESIS: The occurrence of these two cases is a statistically unusual event--a 'cluster'. There are several plausible explanations for this cluster. First, this may be a chance occurrence, and the tumours may have no causative factors in common. Second, the cause may be radiation exposure. A connection to occupational radiation exposure is biologically plausible, but risk assessment suggests that it is unlikely that this effect would have been observable in the small population of Ontario cardiologists. CONCLUSIONS: Initiation of brain tumours during cardiac fluoroscopic procedures is plausible. Physicians are reminded to practise radiation safety methods during fluoroscopic procedures. The diagnosis of two additional brain tumours in Canadian interventional cardiologists during the past 10 years would confirm the occupational causation theory. The author invites physicians to report knowledge of the diagnosis of brain tumours in Canadian cardiologists to the author or to the editors. PMID- 9854521 TI - Killer dreams. AB - Emotional stress is a recognized trigger for coronary artery spasm. An association between dreams and sudden death is described in folklore and medical history, and originates from the common experience of being awakened by vivid, frightening dreams, with racing pulse, cold sweats and other physiological responses associated with intense distress. Intense alterations in autonomic activity during dreaming can have dire consequences in patients with cardiovascular disease. Four patients with no evidence of underlying coronary artery disease, where emotional stress produced by nightmares or 'deadly dreams' caused coronary artery dissection in two and vasospasm in the other two, leading to life-threatening cardiac events, are presented. A possible mechanism is speculated. PMID- 9854522 TI - Hypercalcemic cardiomyopathy associated with primary hyperparathyroidism mimicking primary obstructive hypertrophic cardiomyopathy. AB - A 72-year-old woman presented to hospital with rapidly progressive dyspnea and chest pain on exertion. Physical findings included a grade 3/6 systolic murmur increased by the Valsalva manoeuvre. Transthoracic echocardiography revealed concentric left ventricular hypertrophy, systolic anterior motion of the mitral valve and critical dynamic outflow tract obstruction. The myocardium was strikingly heterogeneous with hyperdynamic left ventricular systolic function. Laboratory findings included severe hypercalcemia secondary to primary hyperparathyroidism. The patient's outcome was unfavourable with nephrogenic diabetes insipidus, pancreatitis, shock, severe acidosis and death. Postmortem examination confirmed the presence of severe concentric left ventricular hypertrophy, a narrowed left ventricular outflow tract and localized endocardial fibrosis of the left interventricular septum. Microscopic findings showed diffuse calcium deposits of the myocardium, coronary arteries, kidneys and lungs. This appears to be the first report of two-dimensional and Doppler echocardiographic findings in hypercalcemic cardiomyopathy mimicking obstructive hypertrophic cardiomyopathy. PMID- 9854523 TI - N-CPAP in the prevention of recurrent intubations and hospitalizations in a patient with refractory congestive heart failure. AB - A 61-year-old woman with ischemic cardiomyopathy continued to have recurrent episodes of respiratory failure secondary to acute pulmonary edema requiring ventilatory support on each occasion, despite undergoing surgical revascularization and mitral valve replacement. These episodes of acute respiratory failure were successfully averted by using nocturnal nasal continuous positive airway pressure (N-CPAP). Following N-CPAP, she was able to stay home for 207 consecutive days. Although well-designed, controlled studies are needed to validate this observation, nocturnal N-CPAP is a viable and cost effective option that may be considered in a select number of patients with end-stage cardiopulmonary disease. PMID- 9854524 TI - The first Dr Robert E Beamish Award. PMID- 9854525 TI - The management of breast lumps. PMID- 9854526 TI - Recommended management for breast lumps. PMID- 9854527 TI - Strangulated Meckel's diverticulum. PMID- 9854528 TI - Soft tissue case 24: arteriovenous fistula secondary to liver biopsy. PMID- 9854529 TI - Musculoskeletal case 1: Lateral discoid meniscus. PMID- 9854530 TI - Scar formation and ligament healing. AB - Ligaments are highly organized, dense, fibrous connective-tissue structures that provide stability to joints and participate in joint proprioception. Injuries to ligaments induce a healing response that is characterized by the formation of a scar. The scar tissue is weaker, larger and creeps more than normal ligament and is associated with an increased amount of minor collagens (types III, V and VI), decreased collagen cross-links and an increased amount of glycosaminoglycans. Studies have shown that certain surgical variables alter the healing of ligaments. Such factors include the size of gap between the healing ligament, ends, the use of motion in a stable joint and the presence of multiple ligamentous injuries. Research on ligament healing includes studies on low-load and failure-load properties, alterations in the expression of matrix molecules, cytokine modulation of healing and gene therapy as a method to alter matrix protein and cytokine production. PMID- 9854531 TI - Item 502: anterior mediastinal mass. PMID- 9854532 TI - Are complication rates for elective primary total hip arthroplasty in Ontario related to surgeon and hospital volumes? A preliminary investigation. AB - OBJECTIVE: To test the hypothesis that complication rates for elective total hip replacement operations are related to surgeon and hospital volumes. DESIGN: Retrospective population cohort study. STUDY COHORT: Patients who had undergone elective total hip replacement in Ontario during 1992 as captured in the Canadian Institute for Health Information database. MAIN OUTCOME MEASURES: In-hospital complications, 1- and 3-year revision rates, 1- and 3-year infection rates, length of hospital stay, and 3-month and 1-year death rates. RESULTS: Surgeons with patient volumes above the 80th percentile (more than 27 hip replacements annually) discharged patients approximately 2.4 days earlier (p < 0.05) than surgeons with volumes below the 40th percentile (less than 9 hip replacements annually) even after adjusting for discharge disposition, hospital volume, patient age, sex, comorbidity and diagnosis. Complication rates requiring hospital readmission and death rates did not differ by surgeon or hospital volume (p > 0.05). CONCLUSIONS: There is no evidence to support regionalization of elective hip replacement surgery in Ontario based on adverse clinical outcomes. Surgeons who perform a large number of total hip replacements are discharging patients earlier than less experienced surgeons, without any-demonstrable increase in complications leading to hospital readmission. The explanation for this observation remains unknown and will require further study. PMID- 9854533 TI - Concordance in communication between surgeon and patient. AB - OBJECTIVES: To examine (1) the capability of using interactive voice response (IVR) system technology for clinical research studies involving assessment of clinician-patient interactions and (2) the concordance of surgeons and their breast cancer patients about the content of a postbiopsy pre-treatment decision meeting. DESIGN: A descriptive comparison of the perceptions of 2 volunteer groups--surgeons and their patients--using interactive voice technology. SETTING: Surgeons' offices. PARTICIPANTS: Twenty-six dyads of surgeons and their patients with newly diagnosed breast cancer. OUTCOME MEASURES: Concordance as determined through a 15-item patient questionnaire and a parallel 11-item surgeon questionnaire addressing surgical and psychosocial aspects of breast cancer treatment. RESULTS: Fifty-four percent to 100% of the 26 dyads indicated concordance about treatment preference, treatment choice, how treatment was chosen, preference for how treatment was chosen, time for discussion about treatment, and discussion about lymph-node removal. Only 27% to 50% of dyads agreed about patient understanding of lymph-node removal, the thoroughness of discussions about adjuvant treatment, the thoroughness of discussion about emotional coping, and the sufficiency of time for the discussion of patient's concerns. In these areas of disagreement surgeons often underestimated the patient's ability to understand and underestimated the patient's perception of the thoroughness of discussions about the psychosocial aspects of the illness (concerns and coping). CONCLUSION: Surgeons and patients demonstrated concordance on their perceptions of the type of treatment desired and needed but were discordant on their perceptions of the degree of patients' understanding about post-treatment and psychosocial issues. PMID- 9854534 TI - The learning curve in laparoscopic inguinal hernia repair for the community general surgeon. AB - OBJECTIVE: To determine the learning curve (number of operations required) to stabilize operating times and complication rates for a general surgeon doing laparoscopic inguinal hernia repair in a community practice. DESIGN: A prospective analysis. SETTING: A 256-bed secondary-care community hospital. PATIENTS: Ninety-eight consecutive patients booked for elective laparoscopic hernia repair on an outpatient basis. INTERVENTIONS: Using the transbdominal preperitoneal approach, 100 operations were carried out to repair 138 groins and a total of 164 separate hernial defects. OUTCOME MEASURES: The number of operations required to decrease operative times and complication rates to a steady level. RESULTS: There were no deaths. There were 5 conversions and 10 admissions, all occurring between the 1st and 46th operations. Two reoperations for reasons other than recurrence were required between the 45th and 55th operations. There were 24 other complications. Complications and surgical times began to level off after 50 operations. The 1 readmission was after the 42nd operation. There were 4 recurrences (2.9% recurrence rate), 2 in each group of 50 operations. Both groups of 2 recurrence occurred within the first 10 operations involving the use of a new stapler. Twenty-two other patients had open hernia repairs because laparoscopy was unsuitable for them. CONCLUSION: The learning curve for laparoscopic inguinal hernia repair in the hands of a general surgeon in community practice who is experienced in open herniorraphy and laparoscopic cholecystectomy is at least 50 operations. PMID- 9854535 TI - General surgeons and clinical ethics: a survey. AB - OBJECTIVE: To determine the preoccupation of general surgeons concerning ethics. DESIGN: A survey by questionnaire. PARTICIPANTS: One thousand members of the Canadian Association of General Surgeons were surveyed through a questionnaire, which inquired about the influence of ethics in their clinical practices. The questionnaire contained 12 questions. There was no recall for those who did not respond. MAIN OUTCOME MEASURES: Responses to questions concerning the sex of the respondents, location of practice, number of years in practice, the presence of hospital support, surgeons' interest in ethical issues, use of autonomy, beneficence, nonmaleficence and justice in solving ethical dilemmas and level of education in clinical ethics. RESULTS: Men made up 95% of the respondents; 64% of respondents had been in practice more than 16 years; 58% came from a community or regional hospital; only 10% had no interest in clinical ethics; only 3% stated that they experienced no ethical problems in their practices; and 52% had no formal education in ethics. CONCLUSIONS: There was general sensitivity for clinical ethics but an evident lack of formal education and of the presence of ethics committees and ethics consultants in many hospitals. PMID- 9854536 TI - Lipoblastoma and liposarcoma in children: an analysis of 9 cases and a review of the literature. AB - OBJECTIVES: To review the experience at a children's hospital of lipoblastoma and liposarcoma and to identify any factors that would differentiate one type of tumour from the other. DESIGN: A retrospective case series. SETTING: British Columbia's Children's Hospital a tertiary-care pediatric centre. PATIENTS: All patients with a pathological diagnosis of lipoblastoma and liposarcoma recorded over 12 years. MAIN OUTCOME MEASURES: The frequency of lipoblastoma and liposarcoma, identified from biopsy specimens of pediatric adipose tumours. The clinical, pathological and cytogenetic variables between lipoblastoma and liposarcoma. RESULTS: One hundred and forty-nine adipose tumours were recorded. Seven (4.7%) were lipoblastomas and 2 (1.3%) were liposarcomas. All tumours presented as asymptomatic, slow-growing, soft-tissue masses. The children with lipoblastoma tended to be younger, but 29% were over 3 years of age. The liposarcoma patients were aged 9 and 14 years. One liposarcoma was of myxoid type and the other was a round cell variant. Karyotypes were reported for 1 lipoblastoma and 1 liposarcoma. The myxoid liposarcoma karyotype was 46,XY,t(12;16)(q13;p11), and the lipoblastoma was reported as 46,XY,der(8)?t(8q;?),+mar. CONCLUSIONS: Lipoblastoma is an unusual childhood neoplasm and liposarcoma is very rare in children. Both tumours may present in a similar fashion, and differentiating them histologically can be difficult. Age cannot be relied upon to accurately predict their behaviour. The tumour karyotype is very helpful in differentiating these neoplasms. PMID- 9854537 TI - Small-bowel necrosis associated with jejunal tube feeding. AB - OBJECTIVES: To report 3 cases of small-bowel necrosis after jejunal tube feeding and to review the literature concerning this condition. DESIGN: A 5-year retrospective review. SETTING: A 560-bed university-affiliated tertiary-care teaching hospital. PATIENTS: Three patients who had bowel necrosis out of 386 who received jejunal tube feedings. RESULTS: The patients experienced small-bowel necrosis as a consequence of jejunal feeding. The ischemic necrosis was preceded by progressive abdominal pain, distension and high nasogastric output. All 3 patients required extensive small-bowel resection. Although survival was rare in previous reports, our 3 patients survived after prompt surgical intervention and small-bowel resection. CONCLUSIONS: Although the death rate for this condition approaches 70%, timely recognition and surgical intervention can save the patient's life. PMID- 9854538 TI - Modified Bentall technique for aortic root replacement: a simplified approach to the short left coronary artery. AB - The short left coronary artery encountered during aortic root replacement with the modified Bentall (coronary button) technique may pose a significant problem for the surgeon. A simple solution entails the placement of a short interposition Dacron graft between the native coronary artery and the aortic graft. This approach has been successfully employed in 2 patients. PMID- 9854539 TI - Metachronous benign solitary fibrous tumours of the pleura (localized "mesotheliomas"): a case report. AB - Solitary fibrous tumours are uncommon pleural tumours that are not related to mesotheliomas. They are typically benign and pedunculated and may grow to massive sizes. Surgical resection is usually curative. Tumour recurrence and metachronous development of multiple tumours are unusual. In this report a patient was treated for 3 benign solitary fibrous tumours of the pleura over 23 years. The authors hypothesize that this represents multifocal tumorigenesis as opposed to local tumour recurrence. The importance of complete surgical excision and lifelong radiographic follow-up are stressed. PMID- 9854540 TI - Bronchobiliary fistula: a case report. AB - Fewer than 20 cases of bronchobiliary fistula have been reported in the literature. In this report a newborn female infant was referred for investigation of gastroesophageal reflux. Upper gastrointestinal endoscopy and 24-hour pH monitoring revealed severe reflux. A fundoplication was carried out. In the immediate postoperative period bile was noticed coming from the endotracheal tube. Bronchoscopy revealed an abnormal opening just to the left of the carina. Fistulography and HIDA scanning confirmed the presence of a bronchobiliary fistula. A right thoracotomy was used to divide the fistula. The child made an uncomplicated recovery. Pathological examination of the excised specimen demonstrated tissue most consistent with an esophageal origin. PMID- 9854541 TI - Spontaneous common iliac vein rupture: a case report. AB - A 68-year-old woman, admitted because of acute lower quadrant abdominal pain but no history of trauma, underwent laparotomy for a suspected ruptured aortic aneurysm. Exploration revealed a 20-mm longitudinal tear in the left iliac vein. The vein was repaired primarily. Her postoperative course was complicated by deep vein thrombosis. Spontaneous rupture of the iliac vein without trauma is rare, but occurs predominantly in healthy white women between the ages of 40 and 80 years. Various causative mechanisms have been described: inflammation of the vessel wall secondary to thrombophlebitis, proximal obstruction of the iliac vein and spontaneous rupture without obstruction or thrombosis. In many cases an increase in intra-abdominal pressure is noted. PMID- 9854542 TI - Imaging before carotid endarterectomy. PMID- 9854543 TI - Guideline for the management of breast lumps. PMID- 9854545 TI - Endovascular irradiation for the prevention of restenosis after percutaneous transluminal angioplasty. PMID- 9854544 TI - Pitfalls in the care of the injured patient. PMID- 9854546 TI - The significance of microcirculatory examinations. PMID- 9854547 TI - Herbimycin-A attenuates ischaemia-reperfusion induced pulmonary neutrophil infiltration. AB - OBJECTIVES: To determine whether pharmacological induction of heat shock proteins (HSPs) by herbimycin-A (a tyrosine-kinase inhibitor) would protect against neutrophil-mediated lung injury in an animal model of lower torso ischaemia reperfusion. MATERIALS AND METHODS: Sprague-Dawley rats were randomised into three groups: the control group underwent midline laparotomy only; the ischaemia reperfusion (IR) group underwent laparotomy and clamping of the infrarenal abdominal aorta for 30 min followed by 2 h of reperfusion; the third group (HerbIR) was pretreated with herbimycin-A 18 h prior to IR insult. Wet to dry lung weight ratio (W:D), bronchoalveolar lavage protein concentration (BALprot), tissue myeloperoxidase activity (MPO) and bronchoalveolar lavage neutrophil count (BALPMN) were measured. Heat shock protein 72 (HSP72) expression in lung, intestine, mesentery and liver was measured using Western immunoblotting. RESULTS: IR resulted in acute lung injury with tissue oedema (W:D) and neutrophil infiltration (PMO, BALPMN). Herbimycin-A, in vivo, induced HSP expression and attenuated neutrophil infiltration (MPO, BALPMN). CONCLUSION: These data indicate that herbimycin-A protects against ischaemia-reperfusion induced pulmonary neutrophil infiltration, possibly by increasing the expression of heat shock proteins. PMID- 9854548 TI - Aortoiliac reconstructive surgery based upon the results of duplex scanning. AB - OBJECTIVE: To evaluate whether duplex scanning can replace angiography in patients operated for aortoiliac obstructive disease. DESIGN: Retrospective. MATERIALS AND METHODS: Between January 1995 and October 1996, 44 patients underwent vascular surgery of the aortoiliac tract. The study population was divided into two groups; patients operated upon the results of duplex scanning only and patients who also underwent angiography prior to surgery. The additional value of angiography and the differences between both groups concerning unexpected peroperative findings, early postoperative failures and the need for additional radiological or surgical interventions in the first three postoperative months were studied. RESULTS: Duplex scan group: 22 patients were operated upon the results of duplex scanning only. In two patients surgical strategy had to be changed. Early postoperative graft occlusion occurred in one case. A haemodynamically significant graft stenosis within 3 months of surgery occurred in one patient. Duplex/angiography group: 22 patients underwent both duplex scanning and angiography. Six patients underwent diagnostic angiography after failed duplex scanning. In 10 patients angiography was part of percutaneous transluminal angioplasty prior to surgery. In six patients angiograms were performed after successful duplex scanning. Angiography failed in two patients and added information in four of 16 patients. Unexpected findings at operation occurred in four patients. Graft stenosis within 3 months was detected in three patients. CONCLUSION: After successful duplex scanning information obtained by angiography has only a limited impact on therapeutic decision-making. In the majority of patients vascular reconstructive surgery of aortoiliac arteries can be planned based on duplex scanning only. PMID- 9854549 TI - Blood product requirements in patients undergoing elective endovascular abdominal aortic aneurysm repair. AB - OBJECTIVE: Endovascular repair (EVR) of abdominal aortic aneurysm (AAA) has many potential advantages, one of which may be reduced blood product requirements. The aim of this study was to compare the current blood product usage in our unit for three types of elective operation: EVR of AAA, open AAA repair and femorofemoral crossover grafts. DESIGN: Prospective data analysis with historical controls. MATERIALS: One-hundred and thirty-two patients undergoing elective EVR of AAA, 35 patients undergoing elective open repair of AAA and 37 patients having femorofemoral crossover grafts. METHODS: Data was collected on the blood product requirements of patients having EVR of AAA, with open AAA repair and femorofemoral crossover graft providing historical controls. RESULTS: There was no difference in the haematological parameters preoperatively between the three groups but postoperatively patients having EVR had a slightly lower haemoglobin than the open group (10.6 g/dl vs. 10.85 g/dl, p = 0.015). The number of patients who received blood transfusion in the EVR group was 82/132 (62%) and the open group 27/35 (77%), p = 0.4. CONCLUSION: Patients undergoing EVR of AAA require blood transfusion in the same numbers when compared to those undergoing open repair. PMID- 9854550 TI - Changes in health-related quality of life after carotid endarterectomy. AB - OBJECTIVES: Carotid endarterectomy (CEA) is intended to prevent stroke and therefore to extend lifespan. Whether CEA also influences health-related quality of life (HRQOL) is largely unknown. This study aimed to assess HRQOL-changes after CEA. DESIGN: Prospective study, data assessment within 1 week before and 3 months after CEA. MATERIALS: Patient classification (n = 70) was based on presenting neurological symptoms (none (24), transient (26), or permanent (20)), patency or occlusion (27%) of the contralateral internal carotid artery and intraoperative shunt requirement (28%). METHODS: HRQOL was investigated with the Sickness Impact Profile (SIP). Analysis of variance was used to adjust for the influence of preoperative differences in functional impairment and comorbidity on the changes found. RESULTS: Preoperative findings showed that the SIP scores of stroke patients and shunted patients were significantly higher (indicating poorer HRQOL) than those of the other patients. No adverse effect of CEA was observed. Analysis of variance revealed that neurological classification was not reflected to HRQOL changes. However, patients with contralateral occlusion showed a significant postoperative improvement (f = 4.99, p < 0.05). CONCLUSIONS: HRQOL improvement after CEA is restricted to patients with occlusion of the contralateral carotid artery. Assessment of outcome of CEA should be related not only to neurological classification, but also to haemodynamic factors such as contralateral occlusion. PMID- 9854551 TI - Suppression of smooth muscle cell proliferation after experimental PTFE arterial grafting: a role for polyclonal anti-basic fibroblast growth factor (bFGF) antibody. AB - OBJECTIVES: To determine the role of polyclonal anti-basic Fibroblast Growth Factor (bFGF) antibody in inhibiting the proliferation of smooth muscle cells after experimental polytetrafluorethilene (PTFE) arterial grafting. MATERIALS: In 14 male inbred Lewis rats (weight 250 mg) a 1 cm long segment of PTFE was interposed at the level of abdominal aorta. Animals were randomised to receive polyclonal anti-bFGF antibody (group A: n = seven animals) or aspecific immunoglobulin (group B: n = seven animals). Anti-bFGF antibody or aspecific immunoglublin were given intraperitoneally at the end of operation, and for the first 2 postoperative days. Animals were sacrificed 7 days after surgery, 24 h after intraperitoneal injection of BromodeoxyUridin (BrdU) to label proliferating smooth muscle cells. RESULTS: One animal in each group died in the immediate postoperative period due to anaesthetic problems. All grafts were patent at the time of sacrifice. BrdU labelling index was statistically higher in the control group B animals at the level of the anastomotic regions (proximal anastomosis: group B 7.9% vs. group A 4.1%. Distal anastomosis: group B 5.1% vs. group A 2.6% p = 0.009) and at the level of PTFE graft (group B 3.8% vs. group A 2.6% p = 0.002), while there was no statistical difference between the control thoracic aorta of the two groups. MAIN CONCLUSIONS: bFGF plays a major role in the proliferation of smooth muscle cells at the level of the anastomoses after arterial PTFE grafting. Agents able to block the action of bFGF may be useful in inhibiting the formation of myointimal hyperplasia. PMID- 9854552 TI - Amlodipine potentiates metalloproteinase activity and accelerates elastin degradation in a model of aneurysmal disease. AB - AIMS: Abdominal aortic aneurysms are characterised by changes in the extracellular matrix of the arterial media, in particular a reduction in elastin concentration. These changes are mediated by increased levels of endogenous matrix metalloproteinases (MMPs). Recently, calcium channel blockers have been shown to increase the proteolytic activity of MMP-2 secreted by vascular smooth muscle cells. It may therefore by hypothesised that calcium antagonists may potentiate the activity of MMPs in aneurysmal disease and thus accelerate AAA expansion. In this study, the ability of amlodipine--a calcium antagonist--to influence elastin degradation, was assessed in a previously described model of aneurysmal disease. METHODS: Porcine aortic segments (n = 8) were pre-incubated in exogenous pancreatic elastase for 24 h prior to culture in standard conditions for 6 days with 10 and 100 micrograms/l amlodipine. Control segments were cultured both with and without amlodipine and without elastase. At the termination of culture MMPs were extracted from the tissue and quantified by a combination of substrate gel enzymography and immunoblotting. The volume fractions of elastin and collagen were determined by stereological analysis of EVG stained sections. RESULTS: Gel enzymography demonstrated significantly increased MMP-9 activity in the amlodipine treated segments, median 4.218 vs. 2.809 arbitrary units (p < 0.01) and this elevated activity was reflected in a significant destruction of medial elastin 27.0 vs. 40.5% (p < 0.05). CONCLUSION: Therapeutic ranges of amlodipine significantly enhanced elastin degradation and potentiated MMP-9 activity within the aortic organ cultures. PMID- 9854553 TI - Accuracy of centrally recorded OPCS codes for vascular surgery in the United Kingdom. AB - AIM: Centrally recorded OPCS codes are based upon district returns. The aim of this study is to determine the accuracy of this system with regard to vascular surgery. METHODS: Prospectively recorded audit data for vascular and endovascular procedures were compared with those obtained from the Department of Health and Welsh Office. Five U.K. hospitals were involved in the study. Data were obtained for the twelve months, 1 April 1994-30 March 1995 (these being the most up to date figures available). RESULTS: The total number of arterial reconstructions based on audit data was 1082. Those recorded by the OPCS codes were 743. This represents a discrepancy of -31.3% (range for the five hospitals: -13.1% to 63.8%). When examining specific codes similar discrepancies were seen. For example, in one hospital 38 AAA repairs were carried out but only two were centrally recorded. However, examination of ICD9 codes (relating to hospital admissions) for that hospital showed that 38 patients with AAA were admitted. A similar wide variation was seen when examining iliac and superficial femoral artery endovascular procedures. Despite the discrepancies of audit and OPCS data, the codes for reconstructions did reflect relative workload of the different hospitals. CONCLUSION: This study shows that there is a marked underestimate of vascular workload when comparing central recorded data with that obtained from local audit. Marked variation is seen in the accuracy of data submitted from different hospitals. PMID- 9854554 TI - The role of infrarenal aortic side branches in the pathogenesis of endoleaks after endovascular aneurysm repair. AB - AIM: To investigate the relation between the number of preoperative patent side branches and the presence or absence of postoperative endoleaks, and to study the fate of patent branches after operation. PATIENTS AND METHODS: Thirty consecutive patients were included. Cine mode viewing of axial CT angiography images was applied to detect infrarenal aortic side branches. The position of side branches relative to the renal arteries, branch patency and run-off pathways were studied. RESULTS: A total of 160 patent side branches were found. All patients had two or more patent side branches. A patent inferior mesenteric artery was found in 22/30 patients (73%). Postoperative CT scans revealed major endoleaks in five patients (16%) and minor endoleaks in eight (27%). There was no significant difference in the number of preoperative patent side branches in patients with a completely thrombosed aneurysm sac (five; range 2-8) compared to patients with postoperative endoleaks (six; range 3-9; p = 0.12). Backbleeding from patent side branches as the sole cause of endoleak was seen in one patient only (3.3%). CONCLUSION: Postoperative endoleaks are not related to the number of preoperative patent side branches. In patients without endoleaks, contrast enhancement of side branches was repeatedly seen in the vicinity of the aneurysm wall. Although close follow up of these branches is warranted, they did not affect the outcome of endovascular aneurysm repair. PMID- 9854555 TI - Compression ultrasonography for false femoral artery aneurysms: hypocoagulability is a cause of failure. AB - OBJECTIVES: False femoral artery aneurysm is an occasional complication of percutaneous cardiovascular radiological procedures. Compression ultrasonography causes thrombosis non-invasively, reducing need for operative intervention. The technique fails in a proportion of cases. Analysis was undertaken to identify causes of failure. DESIGN: Prospective open study. MATERIALS AND METHODS: Patients presenting with false femoral artery aneurysm since 1984 were identified from a computerised database (BIPAS). Since 1993 compression ultrasonography has been performed as first line treatment according to a standard protocol. Prospectively collected ultrasonographic data and case notes were reviewed to identify causes of failed compression. RESULTS: False femoral artery aneurysm occurred as a complication in 32/26,687 (0.12%) cardiovascular radiological procedures. Eighteen aneurysms were treated by compression. The technique was successful in 11/18 (61%) cases but primary failure occurred in seven cases. Six out of seven had bleeding abnormalities (Chi-squared analysis with Yates correction 10.55, p = 0.0012), four were anticoagulated and compression was subsequently successful following reversal of warfarin therapy in three of these patients. In 4/18 cases surgical repair was necessary. CONCLUSION: Compression ultrasonography is an effective treatment of false femoral aneurysms, however, hypocoagulability is a significant cause of failure. For patients in whom anticoagulation cannot be reversed, primary surgical repair should be considered. PMID- 9854556 TI - Estimates of distance by claudicants and vascular surgeons are inherently unreliable. AB - Claudication distance and maximum walking distance are both measures of disability. Estimates of the distances which a patient with intermittent claudication can walk are commonly used by vascular surgeons as surrogate measures of handicap. We studied how accurately distance was estimated by 70 patients with intermittent claudication, and by 100 British vascular surgeons. Patients and vascular surgeons estimated actual distances of 30 m and 34 m, respectively. Estimates were 46 m median (range 2.7-402 m) for claudicants, and 46 m median (range 15.2-186 m) for vascular surgeons, median over estimates of 52% and 34% respectively. Claudicants' errors can easily be eliminated by accurate and appropriate measurement of claudication and maximum walking distances but interpretation of the data by surgeons is dependent on their own ability to estimate distance. Flawed perceptions by patients and their surgeons of the disability of reduced walking distance illustrates the need for reliable measures of handicap on which to base therapeutic decisions in patients with intermittent claudication. PMID- 9854557 TI - The infrarenal aortic diameter in relation to age: only part of the population in older age groups shows an increase. AB - OBJECTIVES: To resolve whether the infrarenal aortic diameter (IAD) continues to increase throughout life; to ascertain the relationship between IAD and age, sex, body size, and smoking status, and to determine whether these factors influence the IAD over the entire range of aortic diameters or only in a proportion. SETTING: Combined cross-sectional data from two population-based screening programmes for abdominal aortic aneurysms (AAA) in Huntingdon (U.K.) and Rotterdam (The Netherlands). METHODS: The antero-posterior diameter of the infrarenal aorta was measured. The influences of age, gender, body size and smoking status were examined. RESULTS: Data were analysed from 3066 women and 8270 men. In men, mean IAD rose from 20.7 mm to 23.5 mm in the older age groups. However, IADs remained constant below the 75th perentile in men and the 85th percentile in women. Similarly only the top 15-25% of the aortic diameters were larger in smokers compared with non-smokers. CONCLUSIONS: The aortic diameter increased with age in only a minority of the population. Furthermore, known risk factors for AAA contributed to aortic dilatation in only the upper tail of the frequency distribution. Thus only 25% of men and 15% of women may be prone to aortic dilatation. PMID- 9854558 TI - Percutaneous transluminal angioplasty in selected patients with severe carotid artery stenoses: the results of a consecutive series of 24 patients. AB - OBJECTIVES: To assess clinically and with duplex scanning the results of percutaneous transluminal angioplasty in a subgroup of patients with carotid artery stenosis. DESIGN: A retrospective study. MATERIALS: Twenty-four patients with focal hemispheric neurological symptoms from a concentric, smooth carotid stenosis more than 70% treated with balloon dilatation. Control investigations were performed with duplex scanning. RESULTS: Angiography after PTA revealed no residual stenosis in 15 patients (62%), a stenosis of less than 50 was seen in seven patients (30%) and PTA was ineffective in two patients (8%). Three patients (13%) had transient ischaemic attacks during the procedure but none had strokes. One patient experienced short lasting paraesthesia of the arm a few hours after the PTA procedure. After a follow-up of a mean of 11 months (1-38) no recurrent symptoms had occurred. Clinical success was thus achieved in 22 of 24 patients (92%). At duplex scanning, 16 (66%) patients had no stenosis, six patients (30%) had a stenosis of 70%, one patient (4%) a stenosis of 80%, and one patient had an occluded carotid artery. CONCLUSION: Our results indicate that PTA in a selected group of patients with carotid artery stenosis is safe and the interim, long-term results are promising. The procedure must be evaluated in a randomised prospective study against surgical treatment. PMID- 9854559 TI - Chyloperitoneum--an infrequent complication of abdominal aortic aneurysm repair. PMID- 9854560 TI - Thromboangiitis obliterans (Buerger's disease) in visceral vessels confirmed by angiographic and histological findings. PMID- 9854561 TI - Treadmills and ergometers. PMID- 9854562 TI - Treadmills and ergometers. PMID- 9854563 TI - The Stresst'er ergometer. PMID- 9854564 TI - The Stresst'er ergometer. PMID- 9854565 TI - Deep venous reflux. PMID- 9854566 TI - Citalopram for depression. PMID- 9854567 TI - Three new drugs for HIV infection. PMID- 9854568 TI - Presidential address: beyond risk groups--a new look at differentiated thyroid cancer. PMID- 9854569 TI - Unilateral total lobectomy: is it sufficient surgical treatment for patients with AMES low-risk papillary thyroid carcinoma? AB - BACKGROUND: Controversy continues regarding the optimal extent of primary thyroid resection in most patients with papillary thyroid carcinoma (PTC), who are at minimal risk of cause-specific mortality (CSM). This study was designed to compare CSM and recurrence rates after either unilateral lobectomy (UL) or bilateral lobar resection (BLR) in patients with PTC considered low risk by AMES criteria. METHODS: Outcome was studied in 1685 patients initially treated during 1940 through 1991 and followed for up to 54 postoperative years (mean, 18 years). One thousand six hundred fifty-six patients (98%) had complete primary tumor resection; 634 (38%) had involvement of regional nodes. One hundred ninety-five patients (12%) had UL; BLR accounted for 1468 (near-total 60%; total thyroidectomy 18%). RESULTS: Thirty-year rates for CSM and distant metastasis were 2% and 3%, respectively. Twenty-year rates for local recurrence and nodal metastasis were 4% and 8%, respectively. There were no significant differences in CSM or distant metastasis rates between UL and BLR (P > .2). After UL, 20-year rates for local recurrence and nodal metastasis were 14% and 19%, significantly higher (P = .0001) than the 2% and 6% rates seen after BLR. CONCLUSIONS: UL was not associated with higher CSM rates, but it was associated with a significantly higher risk of locoregional recurrence. Thus BLR probably represents a preferable initial surgical approach to patients with low-risk PTC. PMID- 9854570 TI - Follicular and Hurthle cell carcinoma: predicting outcome and directing therapy. AB - BACKGROUND: Follicular thyroid cancer is a heterogeneous disease including follicular and Hurthle cell and tumors with and without vascular and major capsular invasion. Analyses of prognosis and risk groups have been criticized for not taking these differences into account. METHODS: Retrospective analysis was done of 240 patients treated from 1940 to 1997. RESULTS: Ninety-two patients without vascular or major capsular invasion followed up for a median of 14 years had no recurrences or deaths. In the remaining 148 patients, 32 had Hurthle cell and 116 had follicular cell carcinoma. Patients with Hurthle cell carcinoma were significantly older (55 vs 47 years; P = .0014). Lymphatic metastases did not influence outcome. Patients who were at high risk by age and sex, metastases, extent, and size had a 20-year survival rate of 36% for follicular and 35% for Hurthle cell carcinoma; patients at low risk had 20-year survival rates of 94% and 89%, respectively, with no significant difference between follicular and Hurthle cell carcinoma. Recurrences were treated successfully in 33% of patients with follicular carcinoma but never cured in patients with Hurthle cell carcinoma. Bilateral versus unilateral operation or radioiodine for ablation did not alter outcome. CONCLUSIONS: Follicular and Hurthle cell carcinoma with minimal capsular invasion behaved benignly. Age and sex, metastases, extent, and size risk criteria differentiate strongly between patients with high-risk and low risk follicular and Hurthle cell carcinoma. Controlling for risk factors, Hurthle cell and follicular cell carcinomas have similar prognoses. PMID- 9854571 TI - Local/regional anesthesia for thyroidectomy: evaluation as an outpatient procedure. AB - BACKGROUND: The purpose of this paper was to review my evolving experience with local/regional anesthesia in an outpatient setting. METHODS: Two hundred three consecutive patients during a 9-year period who chose to undergo thyroid operation under regional/local anesthesia were reviewed. Early discharge was offered to patients who were observed for 6 hours without neck swelling and who had no surgical reasons for delaying discharge. RESULTS: In group A there were 2 patients who were given inhalation anesthesia during operation compared with none in groups B and C. The average length of stay in group A was 0.49 days, 0.55 days in group B, and 0.24 days in group C. Eighty-five percent of the patients whose operation began before 1300 hours were discharged within 6 hours versus only 50% of those operated on later in the day. Forty-seven percent of patients in group A, 65% of group B, and 77% of patients in group C were discharged within 6 hours of operation. On the basis of previous experience with general anesthesia, discharge time is not significantly influenced by the type of anesthesia chosen. There were no readmissions to the hospital, but 2 episodes of postoperative bleeding required reoperation. Survey showed that 95% of patients rated the level of pain equivalent or less severe than dental procedures under local anesthesia, and all patients would choose local again. CONCLUSIONS: These data suggest that thyroidectomy can be performed with the patient under local/regional anesthesia, with low morbidity and high patient satisfaction. Most patients can be discharged within 6 to 8 hours, and these discharges were not associated with readmissions. PMID- 9854572 TI - Case-control study on symptoms and signs of "asymptomatic" primary hyperparathyroidism. AB - BACKGROUND: Symptoms, signs, and treatment of mild primary hyperparathyroidism (HPT) are controversial. METHODS: One hundred two patients with HPT and matched controls were recruited from 5202 females attending population-based mammography screening at age 55 to 75 years. Patients' total serum calcium averaged 10.40 +/- 0.564 mg/dL and intact serum parathyroid hormone 58 +/- 33 ng/L. All patients lacked knowledge of their disease. Questions revealed traditional symptoms of HPT in 24% of cases and 43% of controls (P = .01). All individuals underwent the same biochemical analyses, bone mass determination, and questionnaires on symptoms, illnesses, medications, and background variables. RESULTS: Patients with HPT had more psychic complaints (P = .03 to .007) of lassitude, fatigue, irritability, and lack of sexual and emotional interests. They had lower bone density in total body, spine, and hip (P = .008 to .0004) and higher serum alkaline phosphatase, cholesterol (very-low-density lipoprotein), triglycerides (total, very-low density lipoprotein), glucose, urate, and hemoglobin values (P = .02 to .0001). Patients visited physicians more often (P = .008) and had more antihypertensive therapy (P = .02). CONCLUSIONS: Mild, "asymptomatic" HPT in patients unaware of their disorder displays significant psychic symptoms, bone loss, and risk factors of cardiovascular disease. PMID- 9854573 TI - Surgical correction of primary hyperparathyroidism improves quality of life. AB - BACKGROUND: The SF-36 health status assessment tool is well suited for measuring the morbidity associated with primary hyperparathyroidism (HPT). The purpose of this study was to test the hypothesis that surgical correction of primary HPT leads to measurable improvement in patient reported functional health status and well-being. METHODS: For the past 4 years patients with primary HPT have been asked to complete the SF-36 and to provide additional demographic and condition specific information for study before operation. They then completed the SF-36 again by mail 2 months and 6 months after operation. RESULTS: One hundred forty patients entered the study through March 1998; 110 patients completed follow-up at 2 months and 82 at 6 months. Marked impairment compared to population norms occurred before operation in 7 of 8 domains of health status. Substantial improvement occurred in 5 of 8 domains of health status at 2 months and in 6 of 8 domains at 6 months. Statistically significant improvement was demonstrated in limitations caused by physical and emotional role function, social function, bodily pain, and vitality. CONCLUSIONS: Successful operation to correct primary HPT significantly improves patient reported functional health status and quality of life. Most improvement is seen within 2 months, but additional improvement in both physical and mental function is seen at 6 months. PMID- 9854574 TI - Primary and reoperative parathyroid operations in hyperparathyroidism of multiple endocrine neoplasia type 1. AB - BACKGROUND: Operation and reoperation for hyperparathyroidism in multiple endocrine neoplasia type 1 (MEN 1) is controversial regarding surgical strategy, preoperative localization, and biochemical indexes of recurrence. METHODS: Fifty patients with MEN 1 with hyperparathyroidism were followed up 2 to 27 years after subtotal (SPX; n = 35) or total parathyroidectomy with forearm autografiing (TPX; n = 15), including 24 who underwent 28 reoperations because of persistent or recurrent hyperparathyroidism. RESULTS: Persistent or recurrent hyperparathyroidism was seen in 66% and 20% of patients after SPX involving extirpation of at least 3 glands and TPX, respectively, and 100% after single gland excision as a primary procedure. After reoperation, hypercalcemia was reversed in 33% of patients by SPX and 61% by intended TPX procedures. All patients received vitamin D substitution after TPX, but restricted thyroid function allowed withdrawal in all but 10 patients (36%). Intact serum parathyroid hormone levels in nongrafted and grafted arms rose with time, but only exceptional ratios localized graft recurrence. Localization of recurrent hyperparathyroidism was achieved with 11C-labeled methionine positron emission tomography. CONCLUSION: MEN 1 hyperparathyroidism has a high risk of recurrence, and operation may include primarily SPX of at least 3 glands or TPX, although the latter includes a higher risk of long-term hypoparathyroidism. Reoperation should involve TPX with recognition of the enhanced recurrence rate in individuals with postoperative hyperparathyroidism. PMID- 9854575 TI - Use of somatostatin receptor scintigraphy to image extrahepatic metastases of neuroendocrine tumors. AB - BACKGROUND: The presence of lymph node metastases significantly influences the modality of treatment in patients with liver metastases of neuroendocrine tumors (NET). Somatostatin receptor scintigraphy (Octreo-Scan, Mallinckrodt-Diagnostica, Petten, the Netherlands) is a method for localization and staging NET. The aim of our prospective study was to evaluate the effectiveness of somatostatin receptor scintigraphy in the identification of extrahepatic tumor spread. METHODS: Thirty five patients with liver metastases of NET were studied over a 5-year period. The presence of NET was confirmed histologically in all cases. To detect extrahepatic metastases or local tumor recurrence, conventional imaging techniques and somatostatin receptor scintigraphy were carried out. RESULTS: In correlation with the findings of conventional imaging methods, somatostatin receptor scintigraphy confirmed liver metastases in all patients. Additionally, 19 of 35 patients (54.2%) had extrahepatic tumor lesions not detected by other imaging techniques. Of those, 15 had extensive abdominal or thoracic lymph node metastases, 3 patients had bone metastases, and in 1 patient with bronchial carcinoid local tumor recurrence was detected. All 19 patients were excluded from further evaluation for liver resection or transplantation and subjected to conservative treatment. The somatostatin receptor scintigraphy sensitivity, confirmed at the time of operation, was 91.6%. In 1 patient, in whom cluster transplantation was performed, somatostatin receptor scintigraphy failed to disclose disseminated carcinosis of the pleural cavity, detected at autopsy (false-negative rate 8.3%). CONCLUSION: In our experience, somatostatin receptor scintigraphy provides a highly sensitive diagnostic method to localize metastases of NET. We recommend somatostatin receptor scintigraphy before liver surgery in every patient with hepatic metastases of NET to identify candidates suitable for resection. PMID- 9854576 TI - Prospective biochemical and scintigraphic evaluation of autografted normal parathyroid glands in patients undergoing thyroid operations. AB - BACKGROUND: Parathyroid autotransplantation has been widely used in thyroid operations. Few studies have assessed the function of the autotransplanted glands. The aim of this study was to evaluate the success rate, pattern of biochemical function, and scintigraphic appearance of parathyroid autografts in patients undergoing thyroid operations. METHODS: Twelve patients in whom 1 parathyroid gland was removed during thyroid operation were included. Glands were fragmented and autografted into individual pockets in the brachioradialis muscle of the nondominant forearm. Parathyroid hormone levels were measured in both arms the day of autotransplantation and 2 weeks and 1, 2, 3, and 6 months after operation. Serum calcium levels were also measured at each interval. Sestamibi scanning was performed 6 months after operation in 7 patients. RESULTS: All patients were women with a mean age of 50 +/- 15 years. Serum calcium levels were normal during follow-up. A gradient of parathyroid hormone level of 1.5 or greater between the autotransplanted and nontransplanted arm was found in 10 patients. Isotope uptake at the site of the parathyroid autotransplant was demonstrated in the 7 patients evaluated. CONCLUSIONS: Biochemical function of autotransplanted normal parathyroid tissue was documented in 83% of the patients. Sestamibi scintiscans were able to identify the autotransplanted tissue. PMID- 9854577 TI - Indications, technique, and efficacy of alcohol injection of enlarged parathyroid glands in patients with primary hyperparathyroidism. AB - BACKGROUND: Percutaneous alcohol ablation of the parathyroid gland (PAAP) has been proposed as an alternative treatment for primary hyperparathyroidism in patients unsuitable for surgery. The current study aimed to determine the (1) selection criteria, (2) associated morbidity, and (3) efficacy of PAAP. METHODS: From 1987 to 1998, 36 patients with primary hyperparathyroidism (mean age 65 years) underwent PAAP. The indications for PAAP were (1) medical comorbidity, (2) technically unsafe reoperative surgery, (3) partial ablation of a single remaining gland, and (4) patient choice. RESULTS: There were no long-term complications. Two patients had temporary recurrent laryngeal nerve injury and 4 had temporary hypocalcemia. Over a median follow-up of 16 months, 12 (33%) of the patients remained eucalcemic. For analysis purposes patients were separated into 2 separate groups: 29 with attempted complete ablation and 7 with partial ablation of a single remaining gland only. Ten of the complete ablation group (34%) remained eucalcemic. In the partial ablation group only 2 remained eucalcemic, but all had adequately controlled serum calcium levels. CONCLUSION: PAAP should be considered for hyperparathyroid patients with excessive reoperative morbidity or prohibitive medical comorbidity or those in whom the intent is to partially ablate a single remaining enlarged gland. In these patients close follow-up of serum calcium is required, and repeat treatments may be necessary because recurrence of hypercalcemia is likely. PMID- 9854578 TI - Secondary hyperparathyroidism is an expected consequence of parathyroidectomy for primary hyperparathyroidism: a prospective study. AB - BACKGROUND: Parathyroidectomy for primary hyperparathyroidism (PHPT) can cause secondary hyperparathyroidism, with increased serum parathyroid hormone (PTH) and normal or low serum calcium concentrations. METHODS: A prospective study investigated 78 consecutive patients who underwent exploration for PHPT. Serum intact PTH and total calcium concentrations were measured the evening after operation and ionized Ca++ the following morning. These levels were reassayed 1 week later. RESULTS: Before operation, the mean PTH level was 138 +/- 15 pg/mL, total calcium concentration was 11.6 +/- 0.1 mg/dL, and ionized Ca++ concentration was 1.44 +/- 0.02 mmol/L. On the night of the operation, the PTH level was 11 +/- 2 pg/mL, and the total calcium concentration was 8.9 +/- 0.1 mg/dL. Fifty-five patients had hypoparathyroidism, with a PTH level less than 10 pg/mL. The day after the operation, the ionized Ca++ level was 1.14 +/- 0.01 mmol/L. One week later, PTH, ionized Ca++, and total serum calcium concentrations returned to normal levels. In 9 patients (12%), PTH levels were increased (98 +/- 16 pg/mL), although ionized Ca++ concentrations were normal (1.18 +/- 0.02 mmol/L), demonstrating secondary hyperparathyroidism. Risk factors for postoperative secondary hyperparathyroidism included older age, symptomatic hyperparathyroidism, higher preoperative PTH and alakaline phosphatase levels, and lower serum phosphorous levels. In 70% of these patients, PTH levels returned to normal in 3 to 12 months. CONCLUSIONS: Secondary hyperparathyroidism occurs in 12% of patients after surgical treatment of PHPT. It is transient, possibly compensating for relative hypocalcemia. PMID- 9854579 TI - Cost implications of different surgical management strategies for primary hyperparathyroidism. AB - BACKGROUND: Controversy exists about optimal management of patients with primary hyperparathyroidism. To date, no studies have explored the cost implications of variation in practice. METHODS: Results from a national survey of endocrine surgeons were combined with results from a survey of endocrinologists and financial data from Medicare. Patterns of use of resources were identified, annual costs for the surgical management of primary hyperparathyroidism in the United States were calculated, and the financial impact of variation in practice was estimated. RESULTS: Survey respondents (n = 109) were experienced endocrine surgeons, performing an average of 33 parathyroidectomies annually. Seventy-five percent of patients undergo localization before initial exploration for primary hyperparathyroidism. In order of preference, these studies were sestamibi (43%), ultrasonography (28%), and sestamibi with single-photon emission computed tomography (26%). Although there is variation in preoperative and postoperative practice, in-hospital costs have the greatest influence on total cost. An estimated $282 million is spent annually in the United States on operations for primary hyperparathyroidism. National health expenditures could range by more than $70 million, depending on whether management strategies involving low or high use of resources are employed. CONCLUSIONS: Substantial variation among endocrine surgeons in the management of primary hyperparathyroidism has important cost implications. Implementation of evidence-based guidelines to optimize clinical and economic performance should be considered. PMID- 9854580 TI - Prospective study of the utility of somatostatin-receptor scintigraphy in the evaluation of patients with multiple endocrine neoplasia type 1. AB - BACKGROUND: Neuroendocrine tumors (NETs) are a potentially lethal component of multiple endocrine neoplasia type 1 (MEN 1). Somatostatin receptor scintigraphy (SRS) can be used to localize NETs and evaluate patients for extraduodenopancreatic disease; its utility in managing MEN 1 is undefined. METHODS: All patients with MEN 1 evaluated by SRS from April 1994 to November 1997 are reported. SRS findings were correlated with other imaging studies and operative findings. RESULTS: Thirty-seven SRS studies were performed in 29 patients with MEN 1. SRS identified occult tumor in 36% (4/11) of patients with only biochemical evidence of NET; 2 patients went on to resection. SRS showed tumor in 79% (15/19) of patients with computed tomography (CT)-demonstrated tumor; 30% (6/20) of the SRS lesions were occult on CT. Conversely, 55% (16/29) of CT-identified lesions were occult on SRS. SRS found distant disease in 21% (6/29) of patients. In patients who had previous operations, SRS found tumor in 40% (4/10) of patients, again with both new positive and false-negative results compared with other imaging. SRS also had 3 important false-positive results, including 1 patient who had laparotomy with no tumor identified. CONCLUSIONS: SRS is useful in identifying otherwise occult NETs in patients with MEN 1 and can substantially alter management. However, SRS also has significant false-positive and false-negative results that demand correlation with other studies. PMID- 9854581 TI - Pancreatic islet cell tumor metastasis in multiple endocrine neoplasia type 1: correlation with primary tumor size. AB - BACKGROUND: Islet cell tumor (ICT) metastasis is one of the potentially lethal outcomes of multiple endocrine neoplasia type 1 (MEN 1). Management of ICT in patients with MEN 1 is controversial; some advocate resection based on biochemical evidence of progression, whereas others use tumor size to predict the risk of metastasis and the need for resection. This study correlates the size of primary ICT with the presence of metastases. METHODS: Forty-eight patients with MEN 1 with ICT, from 34 kindreds followed up in our multiple endocrine neoplasia program, were evaluated; 43 of the 48 have been explored for ICT. Metastases to the lymph nodes and liver were documented. RESULTS: Thirty-three percent of patients with pancreatic tumors less than 1 cm in greatest diameter had metastatic disease at surgery and in follow-up, whereas 34.8% of patients with tumors greater than 2 cm in diameter had metastases to lymph nodes or liver. The 2 patients with liver metastases each had primary tumors greater than 2 cm. Follow-up revealed subsequent metastasis in 1 patient. CONCLUSIONS: The size of primary tumors in MEN 1 does not correlate with metastatic potential. This is not a good criterion for exploration. Continued follow-up of these patients will be necessary to define the effect of operation on the course of ICT in MEN 1. PMID- 9854582 TI - Vasoactive intestinal polypeptide secreting islet cell tumors: a 15-year experience and review of the literature. AB - BACKGROUND: Vasoactive intestinal polypeptide (VIP)-secreting tumors of the pancreas represent a rare subtype of pancreatic islet cell tumors with an estimated incidence of 0.2 to 0.5 per million per year. We provide data on a relatively large series of patients with VIP-secreting tumors and review current literature regarding this specific entity. METHODS: A retrospective review was performed of all patients with VIP-secreting tumors of the pancreas treated from 1977 to 1992 at our institution. Presenting signs, symptoms, mode of diagnosis, extent of disease, surgical resectability, tumor size, treatments, hormone levels, and survival were assessed. RESULTS: Eighteen patients were identified, 9 male and 9 female. Ages ranged from 23 to 74 years (mean 51 years). Secretory diarrhea was the most common symptom, occurring in 16 of 18 patients (89%). The most common tumor location was the tail of the pancreas (9 patients). Fourteen patients (78%) had liver metastasis at diagnosis. Curative resections were attempted in only 5 patients (28%). The mean survival was 3.6 years with the longest disease-free survival being 15 years and longest overall survival 15 years. CONCLUSIONS: VIP-secreting tumors are extremely rare entities and usually metastatic at the time of diagnosis. Despite advanced disease, these patients can have extended survival. PMID- 9854583 TI - Operative management of islet-cell tumors arising in the head of the pancreas. AB - BACKGROUND: Pancreatic islet cell tumors (ICTs) can be treated by enucleation or pancreatic resection. We reviewed our experience with ICTs in the head of the pancreas to define indications for enucleation versus pancreaticoduodenectomy. METHODS: Between January 1982 and December 1997, 48 patients underwent surgical resection for presumed ICTs of the pancreatic head. Of these, 18 were found on pathologic examination to be disease in a lymph node. Thirty patients had 32 true pancreatic head ICTs. We reviewed the operative results and postoperative courses in these patients. RESULTS: Mean diameter of the ICTs was 15 mm. Twenty-seven patients (90%) underwent successful enucleations. Three patients failed enucleation and underwent pancreaticoduodenectomy. There was no operative mortality. The median time to regular diet was 7 days. The median time to removal of all drains was 22 days. The most common complication was pancreatic fistula (15%). No patient required reoperation for treatment of a fistula. CONCLUSION: Most ICTs of the pancreatic head can be removed with enucleation, preserving pancreatic tissue and avoiding the morbidity of pancreaticoduodenectomy. The primary indication for pancreaticoduodenectomy is not the size of the lesion but its proximity to the pancreatic duct. PMID- 9854584 TI - Carcinoids of unknown origin: comparative analysis with foregut, midgut, and hindgut carcinoids. AB - BACKGROUND: Carcinoids are rare neuroendocrine tumors typically arising in the gastrointestinal tract. A significant percentage of these tumors present as metastatic disease of unknown primary site. The aim of this study was to better define the functional and clinical characteristics of carcinoids of unknown primary (CUP) site. METHODS: This study examines the hormonal activity, clinical characteristics, and survival of 434 patients with carcinoids originating in the foregut, midgut, hindgut, or unknown location. The 143 patients with CUP were compared with the other groups with regard to presenting characteristics, diagnostic tests and therapeutic modalities used, hormonal activity, and survival. RESULTS: The hormone levels (urinary 5-hydroxyindoleacetic acid and serotonin, serum and platelet serotonin) of CUP were not significantly different from midgut carcinoids with metastatic disease. Although survival with CUP was shorter than with carcinoids with identified primaries (10-year survivals of 22% vs 62%, 50%, and 48% for foregut, midgut, and hindgut, respectively), the survival curve for CUP was quite similar to that of patients with midgut carcinoids with distant disease (10-year survival of 22% vs 28%). CONCLUSIONS: CUP are similar to midgut carcinoids presenting with metastatic disease with regard to hormone production and survival. Like other carcinoids, CUP can be an indolent disease process with gradual progression over decades. PMID- 9854585 TI - A novel cytotoxic agent for human carcinoid tumors. AB - BACKGROUND: Conventional adjuvant therapy for advanced carcinoid tumors remains disappointing; novel therapeutic agents are needed. We have shown previously that inhibiting polyamine biosynthesis with alpha-difluoromethylornithine (DFMO) slows the growth of carcinoid tumors. However, the clinical utility of DFMO has been limited by its cytostatic property. Synthetic polyamine analogs such as 1,19 bis(ethylamino)-5,10,15-triazanonadecane (BE-4-4-4-4) appear to be cytotoxic against several human tumors. The purpose of our study was to determine whether BE-4-4-4-4 is a more effective antiproliferative and cytotoxic agent than DFMO on human carcinoid (BON) cells in vitro. METHODS: BON cells were treated with either 5 mmol/L DFMO, 0.5 to 10 mumol/L BE-4-4-4-4, or vehicle (control). Ornithine decarboxylase activity was determined by the rate of 14CO2 production, and intracellular polyamine levels were determined by chromatography. Cell number and viability were determined by Coulter counter and trypan blue exclusion, respectively. RESULTS: BE-4-4-4-4 inhibited ornithine decarboxylase activity and depleted all 3 polyamines. BE-4-4-4-4 decreased cell numbers by 81% compared with control and 27% compared with DFMO. BE-4-4-4-4 also induced a 2-fold increase in cell death compared with control or DFMO. CONCLUSIONS: BE-4-4-4-4 is cytotoxic and more effective than DFMO in inhibiting growth of BON cells. Polyamine analogs such as BE-4-4-4-4 may be effective adjuvant therapeutic agents for advanced carcinoid tumors. PMID- 9854586 TI - Endoscopic parathyroidectomy: report of an initial experience. AB - BACKGROUND: Preoperative localization of parathyroid lesions and intraoperative quick parathyroid hormone (PTH) assay have been proposed to minimize the extent of operation in primary hyperparathyroidism. To this purpose, endoscopic procedures have been introduced recently. METHODS: During a period of 13 months, 39 of 65 consecutive patients with primary hyperparathyroidism were selected for endoscopic parathyroidectomy on the basis of the following criteria: preoperative echographic diagnosis of a single adenoma, absence of nodular goiter, and no prior neck operations. Unilateral neck exploration and excision of the adenoma was performed through a gasless procedure combined with intraoperative PTH measurements. Mean follow-up after the operation was 7 months (range 1 to 13 months). RESULTS: Thirty-nine parathyroid adenomas were removed; the mean diameter was 21 mm (range 5 to 30 mm). The mean operative time was 65 minutes (range 30 to 180 minutes). In all cases PTH concentration decreased significantly. Patients who underwent endoscopic parathyroidectomy had less postoperative pain compared with patients who underwent conventional hemithyroidectomy. At follow-up, serum calcium and PTH levels were normal in all cases. CONCLUSIONS: Endoscopic parathyroidectomy proved to be a feasible surgical procedure that can be performed in an acceptable operative time with an excellent cosmetic result. The gasless approach avoided any emphysema. PMID- 9854587 TI - Postoperative hypocalcemia in patients who did or did not undergo parathyroid autotransplantation during thyroidectomy: a comparative study. AB - BACKGROUND: Permanent hypoparathyroidism is a recognized complication of thyroidectomy. Apart from preservation of parathyroid glands in situ by meticulous dissection, parathyroid autotransplantation (PTHAT) has been performed increasingly to avoid permanent hypoparathyroidism. METHODS: From January 1995 to October 1997, PTHAT was performed routinely for devascularized or inadvertently removed glands in 98 (36%) of 271 patients undergoing thyroidectomy. Potential risk factors and the impact of PTHAT on postoperative hypocalcemia were studied. RESULTS: Postoperative hypocalcemia occurred in 40 patients (14.8%), whereas 5 patients (1.8%) had permanent hypocalcemia during a median follow-up of 9 months. The incidence of transient hypocalcemia (n = 35) was higher in patients who underwent PTHAT (21.4%) than in patients who did not undergo PTHAT (8.1%) (P < .01). Permanent hypocalcemia occurred only in patients who did not undergo PTHAT. None of the 21 patients who had postoperative hypocalemia after PTHAT had permanent hypoparathyroidism compared with 26% (5/19) of patients who did not undergo PTHAT (P = .018). When the resected thyroid gland was examined for parathyroid tissue, the incidence of positive identification was higher in patients who did not undergo PTHAT (13%) than in patients who did undergo PTHAT (4%) (P = .015). CONCLUSIONS: Patients with postoperative hypocalcemia after PTHAT have virtually no risk of having permanent hypoparathyroidism. A more careful examination of the resected thyroid tissue can help to identify inadvertently removed parathyroid glands for autotransplantation. PMID- 9854588 TI - Minimally invasive radioguided parathyroidectomy in the reoperative neck. AB - BACKGROUND: Operations for hyperparathyroidism (HPT) in a previously operated neck present a significant challenge and carry much higher morbidity rates than first-time operations. Our extensive experience with minimally invasive radioguided parathyroidectomy (MIRP) for first-time surgery for HPT has shown this method to be a directed approach to the offending adenoma, suggesting that the technique could be used to minimize reoperative neck surgery as well. METHODS: Over an 11-month period 24 consecutive patients with primary HPT who had undergone at least one previous neck operation were referred for re-exploration. All patients underwent preoperative sestamibi scanning; 21 localized sufficiently to undergo MIRP. RESULTS: All patients were cured after reoperation. Eighteen patients underwent MIRP under local anesthesia as outpatients; 3 MIRPs were done under general anesthesia. Average total operative time was 44 minutes, average incision length was 3.0 cm +/- 0.2 cm. Nineteen of the procedures were completed without any frozen sections. There were no complications. CONCLUSION: MIRP is extremely effective in patients with HPT who have undergone previous neck exploration for parathyroid or thyroid disease. The technique allows for such a directed dissection that smaller incisions and local anesthesia in an outpatient setting are routine. PMID- 9854589 TI - Decreased expression of calcium-sensing receptor messenger ribonucleic acids in parathyroid adenomas. AB - BACKGROUND: The set point for parathyroid hormone (PTH) secretion is increased in patients with primary hyperparathyroidism, possibly because of receptor defect(s). A decreased expression of calcium receptor (CaR) messenger ribonucleic acid (mRNA) and protein and a decreased expression of the putative calcium sensing CAS (gp330/megalin) protein have been demonstrated in parathyroid adenomas. METHODS: Expression of CAS mRNA was studied in matched pairs of adenomas and adenoma-associated biopsy specimens from normal parathyroid glands from 15 patients with sporadic primary hyperparathyroidism. Cryostat sections were hybridized with an oligonucleotide complementary to CAS mRNA, rinsed, air dried, and exposed to x-ray film for semiquantification of radioactivity. RESULTS: Expression of CAS mRNA in the adenomas was lowered significantly to 25% (median; range 9% to 80%) of that of the corresponding biopsy specimens of normal parathyroid glands. No correlation was seen between CAS mRNA in the adenoma and preoperative serum calcium levels, PTH level, or weight of the adenoma. The levels of CAS mRNA were significantly lower than those observed previously for CaR mRNA. There was no significant correlation between the levels of CAS and CaR mRNA. CONCLUSIONS: Lowered levels of receptors sensing extracellular calcium (CaR and CAS) probably contribute to the increased set point for PTH secretion in primary hyperparathyroidism. PMID- 9854590 TI - Overexpression of TTF-1 and PAX-8 restores thyroglobulin gene promoter activity in ARO and WRO cell lines. AB - BACKGROUND: In anticipation of developing gene therapy against thyroid carcinoma we created an expression vector using the thyroglobulin (Tg) gene promoter. The inhibition of both Tg and thyroid-specific transcription factor (TTF-1 and PAX-8) gene expression, however, has been well documented in thyroid carcinomas. We therefore examined the effects of overexpression of TTF-1 and PAX-8 on Tg gene promoter activity in the human thyroid carcinoma cell lines, ARO (anaplastic) and WRO (follicular). METHODS: ARO, WRO, and nonthyroid cells were transfected with an expression vector in which beta-galactosidase (beta-gal) is driven by the Tg gene promoter (beta-gal). Tg, TTF-1, and PAX-8 gene expression were also examined by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: ARO and WRO exhibited decreased gene expression of Tg, TTF-1, and PAX-8. Transfection with TG -gal alone exhibited minimal beta-gal expression, whereas cotransfection with TTF 1 and PAX-8 resulted in markedly increased expression. There was no evidence of beta-gal expression with or without TTF-1 and PAX-8 in nonthyroid cells. CONCLUSIONS: Weak Tg gene promoter activity in ARO and WRO is associated with decreased expression of transcription factors TTF-1 and PAX-8 but can be restored with their overexpression. This model may serve as a template on which to further develop cell-specific gene therapy against thyroid carcinoma. PMID- 9854591 TI - The variable penetrance and spectrum of manifestations of multiple endocrine neoplasia type 1. AB - BACKGROUND: Some experts maintain that (1) > 90% of patients with multiple endocrine neoplasia type 1 (MEN1) are first seen with hyperparathyroidism (HPTH) so that routine screening for other features is unnecessary and (2) MEN1 has > or = 94% penetrance by age 50 years. METHODS: We constructed a regional registry of patients with or at risk for MEN1 and examined phenotypic profiles in 34 patients. MEN1 was defined as (1) endocrinopathy of 2 of the 3 principal related tissues (parathyroid, gastrointestinal endocrine, pituitary) or (2) 1 such feature plus a first-degree relative with MEN1. RESULTS: The initial feature of MEN1 was HPTH in 50%, pituitary tumor in 18%, and gastrointestinal endocrine tumor in 32% of patients, with overall incidences of 82%, 65%, and 74%, respectively. HPTH developed by age 50 years in 73% of patients and by age 70 years in 83%. Penetrance of MEN1 at age 50 years was 82%. Associated features included renal (1) and rectal (1) cancer, malignant thymic carcinoid (1), and malignant pheochromocytoma (1). CONCLUSIONS: Expression of MEN1 can vary considerably from established patterns. In our geographic region HPTH does not routinely precede other features of MEN1 and cannot be used to distinguish affected patients among those at risk. MEN1 can be inapparent until late in life and may be significantly underdiagnosed. PMID- 9854592 TI - Unknown primary cancer presenting as an adrenal mass: frequency and implications for diagnostic evaluation of adrenal incidentalomas. AB - BACKGROUND: Fine-needle aspiration biopsy to identify adrenal metastasis from an occult primary malignancy has been recommended as part of the evaluation of the patient who presents with an incidentally discovered adrenal mass. This recommendation was assessed by examining the frequency of adrenal involvement in patients with suspected unknown primary cancer. METHODS: Data from 1715 patients referred for evaluation of suspected unknown primary cancer were retrospectively reviewed. RESULTS: Of 1639 patients found to have cancer, the adrenal gland was identified as a site of involvement at presentation in 95 (5.8%). Involvement was limited to the adrenal gland in 4 patients (0.2%). All 4 patients had large (> or = 6 cm) adrenal tumors, 3 of 4 had bilateral involvement, and all had symptoms that otherwise mandated evaluation for an occult malignancy; none had a true adrenal incidentaloma. CONCLUSIONS: Although cancer of an unknown primary site occasionally involves the adrenal gland, metastatic cancer presenting as a true adrenal incidentaloma is extremely rare. Therefore, in the absence of a history of prior malignancy or symptoms, physical examination findings, radiographic findings, or laboratory findings suggestive of an occult malignancy, we do not recommend fine-needle aspiration biopsy as part of the diagnostic evaluation of the patient who presents with a unilateral adrenal mass. PMID- 9854593 TI - Telomerase activity in the differentiation of benign and malignant adrenal tumors. AB - BACKGROUND: Telomerase is an RNA-dependent DNA polymerase that extends the ends of chromosomes by synthesizing the 6 oligonucleotide repeat TTAGGG and thus serves as a marker for cellular immortality. Although absent in most adult somatic tissues, telomerase activity is present in stem cells and is reactivated in nearly all primary human malignancies. In this study we sought to determine whether tumors of the adrenal glands contain telomerase activity and whether telomerase activity can be used to differentiate benign and malignant tumors of the adrenal glands. METHODS: Tissue was obtained from 23 specimens at adrenalectomy. Adjacent normal adrenal tissue was obtained for control. All specimens were rapidly frozen and stored at -80 degrees C until assay. Telomerase activity was determined by the telomeric repeat amplification protocol (TRAP). RESULTS: Telomerase activity was present in 5 of 23 (22%) of the adrenal tumors. All 3 malignant tumors were strongly TRAP positive. There was a single cortical adenoma that had very weak telomerase activity. The single TRAP-positive tumor of the adrenal medulla was a ganglioneuroma. CONCLUSIONS: Benign adrenal tumors infrequently contain telomerase activity, whereas telomerase reactivation appears to be common in malignant tumors of the adrenal glands. These data suggest that determination of telomerase activity may offer a novel way to facilitate the differentiation of benign and malignant adrenal tumors. PMID- 9854594 TI - Essential hypertension: first reason for persistent hypertension after unilateral adrenalectomy for primary aldosteronism? AB - BACKGROUND: Despite cure of primary aldosteronism by surgical resection, hypertension persists postoperatively in 30% to 50% of patients. The aim of this study was to determine factors influencing long-term outcome of blood pressure after unilateral adrenalectomy for primary aldosteronism. METHODS: Records of 100 patients who underwent unilateral adrenalectomy for primary aldosteronism from 1970 through 1997 were reviewed. Patients were distributed in 2 groups according to whether blood pressure was normal (criteria of World Health Organization). Clinical, biochemical, and pathologic data were compared. RESULTS: All patients were biochemically cured. Blood pressure was normal in 56 patients and improved in 44 (mean follow-up, 69 and 59 months). Persistent hypertension correlated with age, known duration and seriousness of preoperative hypertension, family history of hypertension, no preoperative response to spironolactone, and contralateral adrenal hypertrophy. Gender, surgical approach, and pathologic findings were not predictive factors of blood pressure outcome. The prevalence of hypertension was almost the same in these postoperative patients as the prevalence of essential hypertension in a random population of the same age. CONCLUSIONS: Early unilateral adrenalectomy allows cure or improvement of hypertension in all patients with primary aldosteronism induced by unilateral excessive source of aldosterone secretion regardless of the pathologic findings. Persistent hypertension suggests that coexisting essential hypertension is present. PMID- 9854595 TI - Noninvasive imaging of insulinomas and gastrinomas with endoscopic ultrasonography and somatostatin receptor scintigraphy. AB - BACKGROUND: Classic morphological techniques are of limited value for imaging endocrine duodenopancreatic tumors, and invasive procedures such as intraarterial stimulation are often used. Two noninvasive procedures, endoscopic ultrasonography (EUS) and somatostatin receptor scintigraphy (SRS), were recently described with promising results. METHODS: In this study we correlated the results of preoperative EUS (n = 34) and SRS (n = 30) with operative findings in patients with histologically proven insulinoma (n = 20) or gastrinoma (n = 21). RESULTS: The sensitivity and positive predictive value (PPV) of EUS were respectively 77% and 94% for pancreatic tumors (insulinomas and gastrinomas), 40% and 100% for duodenal gastrinomas, and 58% and 78% for metastatic lymph nodes. The sensitivity and PPV of SRS for insulinoma were 60% and 100%, respectively. In patients with gastrinoma, the sensitivity and PPV of SRS were respectively 25% and 100% for pancreatic gastrinomas, 72% and 100% for duodenal gastrinomas or periduodenal metastatic lymph nodes, and 67% and 80% for liver metastasis. In patients with multiple endocrine neoplasia, neither one of the two techniques detected all tumors. Overall sensitivity of combined EUS and SRS was 89% for insulinoma (n = 9) and 93% for gastrinoma (n = 14). CONCLUSIONS: EUS and SRS for gastrinomas and insulinomas should be considered as the initial preoperative imaging procedures and may render invasive procedures unnecessary for most patients. PMID- 9854596 TI - Surgical treatment of localized gastrinoma within the liver: a prospective study. AB - BACKGROUND: Studies demonstrate that liver metastases of gastrinoma significantly reduce survival. METHODS: Since 1982 we have prospectively studied 213 patients with Zollinger-Ellison syndrome. For this report the results of surgery for localized liver gastrinoma were analyzed. RESULTS: Zollinger-Ellison syndrome was diagnosed biochemically in all patients and acid output was controlled with medications. Imaging studies demonstrated liver gastrinoma in 69 patients (32%). Fifty-two had diffuse unresectable disease, whereas 17 (10%) had localized disease. All patients with localized liver gastrinoma and 2 patients with diffuse disease who needed surgery are the subject of this report. Major hepatic lobectomy was performed in 10 patients and wedge resections in 9. Three patients had apparent liver primary gastrinomas and 16 had metastatic disease. Seventeen of 19 patients were able to have all identifiable gastrinoma resected. Extrahepatic tumor was also removed at the same procedure. Extirpation of liver gastrinoma required hepatic lobectomy in 10 patients and wedge resections in the others. Five-year survival was 85%. Five of 17 completely resected patients (29%) remained disease free. CONCLUSIONS: Resectable localized liver gastrinoma is rare. Primary liver gastrinomas can occur. Surgical resection of localized liver gastrinoma provides a cure rate similar to that of extrahepatic gastrinoma and an excellent long-term survival. PMID- 9854597 TI - Pancreatic neuroendocrine tumors associated with von Hippel Lindau disease: diagnostic and management recommendations. AB - BACKGROUND: von Hippel Lindau disease (VHL) is an inherited syndrome characterized by tumors of the kidney, adrenal, central nervous system, and pancreas. The incidence and natural history of pancreatic neuroendocrine tumors occurring in VHL are not known. METHODS: From December 1988 through November 1997, 256 patients with VHL were screened with imaging studies, and these data were reviewed from a prospective database. RESULTS: Thirty (12%) of 256 patients had solid pancreatic lesions consistent with neuroendocrine tumors. Fourteen patients underwent resection, and 4 with metastases on imaging studies underwent biopsy only. Of the 14 patients who underwent resection, 11 remain free of disease, 2 have experienced recurrence, and 1 has died of unrelated causes (mean follow-up, 25 months; range, 3 to 73 months). The size of the primary tumor (median, 5 cm; range, 3 to 8 cm) in patients with liver metastases was significantly larger than the size of the primary tumor (median, 2 cm; range, 1 to 5 cm) in patients without liver metastases (P = .0013). CONCLUSIONS: Solid pancreatic lesions were detected in 12% of patients with VHL. Larger primary tumors were associated with liver metastases. Pancreatic imaging to identify neuroendocrine tumors and resection when they reach 2 to 3 cm may prevent the development of hepatic metastases. PMID- 9854598 TI - Analysis of factors associated with long-term (five or more years) cure in patients undergoing operation for Zollinger-Ellison syndrome. AB - BACKGROUND: Only 30% to 40% of patients with Zollinger-Ellison syndrome (ZES) undergoing operation with curative intent have biochemical cures. The purpose of this analysis was to identify perioperative factors associated with long-term (> or = 5 years) biochemical cures. METHODS: From December 1981 to September 1997, 128 patients with potentially curable ZES underwent 152 abdominal explorations with resection of all identifiable disease. Of these, 31 outcomes were identified with documented cures at > or = 5 years from operation and were compared with outcomes of 110 patients who were not cured. Univariate and multivariate statistical analyses were performed. RESULTS: The results of a normal immediate postoperative fasting serum gastrin and secretin stimulation test were significantly and independently correlated with 5-year cure (P2 = .005 and .0099, respectively). A diagnosis of multiple endocrine neoplasia type 1 was significantly inversely correlated with cure on univariate analysis (P2 = .027). Gender, age, duration of symptoms, results of diagnostic tests, or results of imaging studies did not correlate with outcome. CONCLUSIONS: All patients with sporadic potentially curable ZES should undergo exploration because outcome is not associated with preoperative tests. Only 5% of patients with ZES and multiple endocrine neoplasia type 1 were cured at 5 years. A normal fasting serum gastrin or secretin stimulation value value immediately postoperatively provides important prognostic information. PMID- 9854599 TI - The association between Crohn disease and the myelodysplastic syndromes. Report of 3 cases and review of the literature. AB - We report 3 patients who developed Crohn disease and myelodysplastic syndrome concurrently and review 9 previously reported cases of this association. Demographic and clinical features, treatment, and outcome are presented from previous reports and our own 3 cases. Of the 12 patients, 8 were men, and the mean age was 68.8 years (range, 28-83 yr). The geographic origin was heterogeneous. Pancytopenia was found in 5 patients, anemia in 5, and bicytopenia in 2. The classification types of myelodysplastic syndrome were variable. The marrow karyotype was reported in 10 patients: it was abnormal in 5, with chromosome 20 abnormalities in 3. Crohn disease and myelodysplasia were diagnosed simultaneously in 5 patients, whereas Crohn disease antedated myelodysplasia in 4 patients (interval, 8 mo-30 yr) and myelodysplasia antedated Crohn disease in 3 patients (interval, 2 mo-4 yr). Signs and symptoms of Crohn disease improved in most patients upon specific medical treatment. However, the outcome of myelodysplastic syndrome was not favorable in 10 of 11 patients in whom it was reported, with 4 deaths due to myelodysplasia-related complications. Notably, the activity of myelodysplasia in 1 of our own patients paralleled the clinical activity of Crohn disease, and hematologic abnormalities of the myelodysplasia resolved upon successful treatment of the Crohn disease, thus strengthening the hypothesis of a pathogenetic link between the disorders. The possibility of myelodysplastic syndrome should be considered in patients with Crohn disease over the age of 50 years who have peripheral blood cytopenias. Likewise, a diagnosis of Crohn disease should be considered in patients with myelodysplastic syndrome who develop gastrointestinal symptoms such as chronic and/or bloody diarrhea. In patients with such an association, the possibility of improvement of the myelodysplasia upon successful therapy of Crohn disease should not be ruled out. PMID- 9854600 TI - Systemic immune reactions to HMG-CoA reductase inhibitors. Report of 4 cases and review of the literature. AB - Four patients with systemic autoimmune disorders, 3 of a serious nature, presented to 1 cardiologist over a 20-month span. In 3 of these cases, an HMG-CoA reductase inhibitor was presumably etiologic, while in the fourth case, the HMG CoA reductase inhibitor might have unmasked the disorder. It would be useful to determine the true frequency of this complication, particularly in older patients not included in most of the statin trials to date. It is well established that autoimmune phenomena and particularly the development of autoantibodies increase with age. The data presented in this report that the group of HMG-CoA reductase inhibitors could be a heretofore poorly recognized etiologic agent. This issue might be addressed by a case-control study looking at the prevalence of statin use in elderly patients with systemic autoimmune disorders and in controls. Until then, the authors advise caution in the use of this class of medications in patient subgroups for whom no clear-cut clinical benefit has yet been proven. PMID- 9854601 TI - A controlled trial of desmopressin and behavioral therapy for nocturnal enuresis. AB - The combination of desmopressin (DDAVP) and behavioral therapy for treatment of nocturnal enuresis was compared with use of each of these modes alone. We randomly assigned 226 enuretic children being treated in primary care clinics of a major medical center in the largest health maintenance organization in Israel into 3 groups: Group A) DDAVP plus behavioral therapy (double-blind); Group B) behavioral therapy plus placebo (double-blind); and Group C) DDAVP alone (open group). DDAVP (20 micrograms/naris) and placebo were administered by intranasal spray. Both pharmacologic and behavioral therapy were initiated after a 2-week observation period and continued for 8 weeks. All patients were followed for 2 months after completion of treatment. A significant reduction in the number of wet nights/week was registered for all 3 groups: 49% in Group A, 45% in Group B, and 19% in Group C. After controlling for confounding factors, no significant difference in effect was noted among the 3 types of treatment during the trial period. However, on follow-up the results for the DDAVP patients were significantly less stable compared with the other 2 groups (p = 0.015). Minor side effects were registered, but none of the participants withdrew from the trial. To our knowledge, this is the largest randomized trial of nocturnal enuresis conducted to date. Our findings suggest that simply discussing the problem with the patient and family leads to improvement, and that behavioral therapy is also beneficial. DDAVP can help, but the relapse rate on discontinuation is high. PMID- 9854602 TI - The phenotype of ostensibly healthy women who are carriers for ornithine transcarbamylase deficiency. AB - Ornithine transcarbamylase (OTC) deficiency is an X-linked disorder of urea synthesis. Among females who carry a mutant OTC allele, there is a wide range of phenotypic variability, ranging from apparent normality to a severe onset and the resulting profound neurologic impairment observed in hemizygous males. This study was designed to define the phenotypic variability of OTC deficiency in ostensibly healthy carrier females and to compare them to noncarrier females from their own and other families. One hundred seventy-five women from 89 families participated in this study. Each completed a mailed questionnaire, allopurinol testing, and fasting plasma amino acid determinations. OTC carrier status was determined by pedigree analysis, allopurinol test results, and/or DNA mutation analysis. Overall, 79 women were identified as carriers of a mutant OTC allele (60 proband mothers, 19 relatives), and 96 women (32 proband mothers, 64 female relatives) were determined to be noncarriers. Comparison of biochemical phenotypes indicated that carriers and noncarriers do not differ in daily urinary creatinine excretion, but that carriers excrete significantly less urea nitrogen and total nitrogen, reflecting their significantly lower historically reported daily protein intake. Carriers had significantly higher levels of fasting plasma glutamine and alanine, and significantly lower levels of citrulline and arginine compared with noncarriers. Carriers and noncarriers reported similar demographic characteristics, anthropometric measurements, level of education, and medical and pregnancy histories. There was no indication of increased incidence of migraine headaches among carriers. Thus, we found no evidence that asymptomatic adult female OTC heterozygotes are at increased risk for previously unidentified health problems apart from an unknown risk for hyperammonemic encephalopathy as occurred in 3 of the carriers in this study. Because these episodes appear to be related to physiologic stress (fracture, parturition), it would seem medically prudent for carriers to be aware of this risk. PMID- 9854603 TI - Remission of primary hyperparathyroidism due to spontaneous infarction of a parathyroid adenoma. Case report and review of the literature. AB - Subclinical infarction of abnormal parathyroid tissue likely occurs relatively frequently, but remission of primary hyperparathyroidism due to spontaneous massive infarction of a parathyroid adenoma is a rare occurrence. We describe a patient who was unique in that spontaneous remission was accompanied by disappearance of the parathyroid adenoma on serial parathyroid scans. We identified 11 other patients in the literature with sufficient documentation of clinical remission of primary hyperparathyroidism due to spontaneous massive infarction of a parathyroid adenoma and review their clinical features as well. Manifestations varied according to the delay in diagnosis, alteration in serum calcium, location, and degree of associated hemorrhage. Ages ranged from 19 to 73 years, and both sexes were affected. Preceding symptoms suggestive of hypercalcemia and/or prior documentation of hypercalcemia were present in 10 cases. Neck pain was present at the time of infarction in 6/12 patients, and a neck mass in 7/12. Only 1 patient died. The initial calcium level done postinfarction was "normal" in 7/12 patients, low in 3/12, and high in 2/12. The parathyroid adenoma was 2 cm or larger in 8/12 patients. In all patients who had parathyroidectomy, histopathology confirmed extensive infarction (bland or hemorrhagic) of the adenoma. The clinical scenarios encountered were 3 hypocalcemic crises, 2 hypercalcemic crises ("acute parathyroid intoxication"), 1 respiratory arrest secondary to a mediastinal hematoma, 2 patients with neck pain as the major complaint, and 4 with no symptoms apart from the premorbid symptoms of hypercalcemia. PMID- 9854604 TI - Cutaneous vasculitis in children and adults. Associated diseases and etiologic factors in 303 patients. AB - Cutaneous vasculitis (CV), a condition characterized by palpable purpura and nonspecific histopathologic findings, presents a diagnostic and therapeutic challenge because it may be a primary disorder or it may be a cutaneous manifestation of another entity, such as systemic necrotizing vasculitis, connective tissue disease, systemic bacterial infection, or malignancy. We studied 303 unselected patients (172 adults and 131 children) with CV to assess the disease associations and etiologic factors, to identify the frequency of primary and secondary CV in different age-groups, and to characterize features that help to distinguish between primary and secondary CV. Of the 131 children, 130 had primary CV: Henoch-Schonlein purpura (HSP) in 116 and hypersensitivity vasculitis (HV) in 14. In contrast, of the 172 adults, only 120 had primary CV: HSP in 39, HV in 70, and essential mixed cryoglobulinemia in 11. CV was a manifestation of systemic necrotizing vasculitis in 23 adults (polyarteritis nodosa in 17, Wegener granulomatosis in 4, and Churg-Strauss syndrome in 2). CV was secondary to other processes in 29 adults: in 20 patients CV was associated with connective tissue disease or another autoimmune or rheumatic disease, in 5 patients CV was a manifestation of severe bacterial infection, especially bacterial endocarditis (4 cases), and in the other 4 patients CV was the presenting symptom of an underlying malignancy. The patients for whom CV was a manifestation of systemic necrotizing vasculitis or secondary to a connective tissue disease, severe bacterial infection, or malignancy had clinical and laboratory data suggestive of the associated disorder. The clinical picture and outcome of primary CV in both children and adults were benign. By contrast, the prognosis of patients with CV in the context of systemic necrotizing vasculitis or secondary to other entities depended on the primary process. Given the different disease association in children and adults, we propose a simple diagnostic workup in children with CV. By contrast the diagnostic approach in adults with CV should be more cautious and the workup more extensive. The early differentiation between primary CV, secondary CV, and CV presenting as a symptom of systemic necrotizing vasculitis, especially in adults, is of paramount importance for an adequate diagnosis and appropriate treatment. PMID- 9854605 TI - [Scanning probe microscopy and medico-biological nanotechnology: history and prospects]. AB - Literature data on the use of scanning probe microscopy for biological samples studying are presented. As an illustration, some results of the authors' investigations of the shark olfactory receptor surface, human lipoprotein of low density and human erythrocyte are given. Perspectives of the medico-biological development of nanotechnologies are evaluated. PMID- 9854606 TI - [Neurulational forms of the pathology of human nervous system]. AB - 47 human embryos aged from 22 days to 6 weeks with various brain anomalies were studied. Space reconstruction of the brain organization is performed in 6 embryos. Neurulation anomalies analysis allowed to establish that human nervous system development is highly unstable between days 21 and 27 after fecundation when active change of the brain shape takes place and non-specific disturbance or delay of nervous cylinders closing results in the development of the nervous system embryonal pathologies. Various forms of the brain embryonal anomalies result from the three following variants of the neurulation disturbances; neurulation halt in the postneuropore zone leads to anomalies of the intermediate and median brain and occipital region of the head; neurulation disturbance in the caudal area of the nervous tube is the cause of spinal cord anomalies. The above anomalies are formed as a result of local neurulation disturbances which are the primary cause of the morphogenetic pathologies in the nervous system development. Therefore, it is justifiable to classify early embryonal anomalies of the nervous system according to their morphological origin and not by the final state of the fetus. PMID- 9854607 TI - [Morphofunctional characteristics of the endosteum in connection with bone remodeling]. AB - Current information on the endost structure at different stages of bone remodelling is given on the basis of the original data and literature. Possible causes and character of the bone cells activity changes during the cycle of remodelling at the decreased functional stimulation on the bone structures and under pathological conditions are discussed. PMID- 9854608 TI - [Morphofunctional disorders of uterine-placenta blood flow in multiple uterine myomas]. AB - Results of the right uterine artery dopplerometry at pregnancy 39-39 weeks and those of subsequent histotopographic and immunomorphologic study of the placental bed in the amputated uteri were compared in 50 pregnant women. Morphofunctional equivalent of a considerable reduction of the uterine-placental circulation depending on the myomatous tissue spread and particularly in rare observations of "the placenta on the node" were found. Pathogenesis of these disturbances is determined by deficiency of the second wave of interstitial cytotrophoblast invasion, by phenomenon of the uterine-placental circulation decrease due to arterial supply of myomatous nodes and local hormonal changes produced by alterations of cellular-tissular correlations in the uterine-placental bed. PMID- 9854609 TI - [Morphology of the lymphoid tissue of the bronchus in healthy persons of various ages, and the effect of smoking]. AB - Bronchoassociated lymphoid tissue (BALT) is studied histologically and morphometrically in 49 autopsy cases of persons of various ages after violent death. Lymphoepithelial follicles typical for BALT of small laboratory animals were not observed in human lungs of healthy persons. BALT structural components are represented by interepithelial lymphocytes, diffusely scattered lymphocytes in the mucous membrane tunica propria, lymphoid agglomerations (focal loose agglomerations in the mucous membrane tunica propria, focal periglandular and dense periductal, dense peribronchial and peribronchiolar agglomerations). According to the morphometrical study, there is BALT hyperplasia at young and adolescent ages and involutive changes at advanced age. Reduction of periglandular lymphoid agglomerations and an increase of the alveolar macrophages number were observed in adult persons who smoked for many years. PMID- 9854610 TI - [Morphological characteristics of lungs in embryos and fetuses in women living in regions contaminated with radionuclides after accident at the Chernobyl power plant]. AB - The possibility of transplacental passage of Cr-90 and Cs-137 as well as their traces appearance in descendants muscles and bones beginning from the 2nd pregnancy trimester (and this was sometimes associated with teratogenic effects) was demonstrated on the abortion material (human embryos and fetuses) from the controlled zones of the Bryansk region. Statistically significant inhibition of the bronchial branching (according to the morphometry of the lung histological section) this indicating the disturbance of the lung prenatal morphogenesis during the "pseudoglandular" developmental stage was observed in the 1st trimester of the pregnancy in the descendants of the irradiated mothers. Lung hypoplasia was observed in some cases in the 2nd and 3rd trimesters of the pregnancy. Activation of lymphoid cells in the fetal lungs and the tendency to the enhancement in them of the ectopic erythroid hemopoiesis was revealed. These processes are considered as a reflection of the compensatory adaptive cellular reaction in the descendants respiratory organs in response to the tissue hypoxia and antigenic stimuli from the organism of the pregnant woman irradiated after the Chernobyl accident. PMID- 9854611 TI - [Cytological characteristics, elemental composition of alveolar macrophages from bronchoalveolar lavage, lung pathology in liquidators of the consequences of the accident at the Chernobyl power plant]. AB - Bronchoalveolar lavage fluid (BALF), broncho- and transbronchial biopsies were examined in Chernobyl wreckers eleven years after the accident. Cytologically and electron-microscopically, there was predominance of alveolar macrophages (AM) with many cytoplasmic foreign bodies containing clear needle-like crystals. Roentgenospectral analysis and laser mass-spectrometry revealed chemical elements which are part of non-active "hot particles" (Al, Si, Ca, Fe) as well as reactor fuel elements and the station constructive materials which are the active part of "hot particles". Programmed BALF allowed to decrease cytosis and the amount of AM containing foreign-body particles as well as concentration of the majority of chemical elements in AM. PMID- 9854612 TI - [Surfactant-dependent atelectasis in pulmonary tuberculosis]. AB - Using mathematic modelling of the secretion function of type II alveolocytes of the right lung intercellular surfactant production in treated and not treated rabbits with multi cavernous tuberculosis of the lung was evaluated. Correlation between its disorders and frequency of development and dissemination of atelectatic changes in the organ was revealed. Surfactant-dependent character of alveoli atelectasis in animals receiving a combination of antituberculosis drugs (isoniazide plus rifampycin plus etambutol), a direct effect of these drugs on membranes of the granular endoplasmic reticulum, formation of secretion granules was demonstrated. Daily intravenous administration of amboxol (25 mg/kg) for 14 days had a protective effect on type II alveolocytes, activated surfactant production, prevented alveoli from atelectasis. PMID- 9854613 TI - [Experimental and clinical pulmonary pneumocystosis]. AB - Rat lungs with hydrocortisone-induced pneumocystosis and a necropsy material from 5 patients with AIDS were studied light- and electron-microscopically. The identity of basic morphological features of lung pneumocystosis development in experiment and humans is demonstrated. Three main periods--latent, pronounced, terminal (pneumocystic pneumonia) are distinguished taking into account the relation of productive and vegetative forms of Pneumocystis carinii, state of the surfactants, type I alveolocytes and inflammation cells. PMID- 9854614 TI - [Electron microscopic study of the liver in chronic viral hepatitis C complicated by alcoholic liver disease]. AB - Combination of HCV-infection and alcoholic liver disease is associated with enhancement of hepatocellular damage (lipid and hydropic degeneration become diffuse), a decrease of hepatocyte necrotic foci and lymphomacrophagal infiltration, an increase of hepatocyte apoptosis and liver fibrosis, absence of lymphoid follicles. An important role in the genesis of these liver alterations is attributed to the combination of viral and alcoholic factors with persistence of hepatitis C virus and liver reticulo-endothelial system depression. PMID- 9854615 TI - [Morphological equivalents of lymphocytic-epithelial interactions in kidney and liver regeneration]. AB - The histo- and ultrastructure of the murine kidney and liver is studied 17 and 48 h, respectively, after unilateral nephrectomy or partial hepatectomy. The number of lymphocytes are found to be increased in these organs after the operation. They come in close contact with hepatocytes and renal tubular epithelial cells forming in some cases gap junctions. It occurs more often 48 h after operation. The number of leukocytes is found to be increased also especially in the regenerating liver in which many hepatocytes with cytoplasm partial necrosis are found. Only single epithelial cells are involved in the destruction in the kidney. Possibility of lymphocyte participation in the morphogenetic function realization is discussed. PMID- 9854616 TI - [Pathological anatomy of alimentary fluorine deficiency in goats]. AB - Chronic alimentary fluorine deficiency was provoked in goats by a semisynthetic diet with fruorine deficiency (less than 0.3 mg/kg dry feed). Teeth caries, emaciation in spite of higher consumption of food as compared to controls, focal inflammatory degenerative changes in the alimentary tract mucous membranes (catarrhal-purulent esophagitis, chronic duodenitis), chronic degenerative changes in the parenchymatous organs were observed. Endocrinopathies, accidental thymus involution, hypothyroid state, destructive changes in the pancreatic insullar cells were also common. PMID- 9854617 TI - [Bacillary generalized angiomatosis in HIV infection]. AB - A unique case of generalized bacillary angiomatosis (BA) in a patient who died of HIV infection is described. Apart from widely spread skin lesions there were also manifestations in the brain, lungs, heart, esophagus and intestine. Gram-negative bacteria were found in the histological sections. Oval and roundish bacteria with a predominantly perivascular location were found electron microscopically in the archives material. PMID- 9854618 TI - [Rupture of atherosclerotic aneurysms of abdominal aorta into the duodenum]. AB - A woman, 77 years old, and two men aged 65 and 70 years died 28-96 hours after sudden beginning of recurrent massive gastrointestinal hemorrhages of clinically unknown origin. Ruptured atherosclerotic abdominal aortic aneurysms with aortoduodenal fistulas were revealed at autopsies. Anatomical predispositions for these rare complications and the causes of the diagnostic mistakes are discussed. PMID- 9854619 TI - [The past, present and future of cell pathology problems]. AB - Three stages of development in cellular pathology are analyzed. Evaluation of important achievements in the past which permitted the advance to the present stage in cellular pathology is given. New methods in experimental pathology resulted in the appearance of a new direction--molecular morphology and molecular cell pathology. The basic features of this direction and new problems to be resolved are formulated. PMID- 9854620 TI - [Biological capabilities of the thymus of human fetus during normal development and during constant exposure to small doses of radiation after the accident at the Chernobyl power plant]. AB - The literature information and the authors's data are presented on the lability of human fetus thymus changes under extremal conditions. In particular, the source of the thymus development is discussed determining its morphogenetic potencies, the sequences of its endocrine and lymphocytopoietic function appearance. Lymphocytes composition and kinetics of their quantity in the process of fetus development and thymus response to the unfavourable conditions of the mother are presented. PMID- 9854621 TI - [The antiparkinson activity of muscarinic antagonists depending on their selectivity for individual m-cholinoreceptor subtypes]. AB - The dependence between the activity parameters of muscarine antagonists in the prevention of haloperidol catalepsy in rats and those in tests characterizing the interaction of ligands and various subtypes of m-cholinoceptors was studied. It was established by constructing the mathematical dependence that blockade of m1 cholinoceptors increases, while that of m2-cholinoceptors reduces the antiparkinsonian activity of the drugs. The activity of the muscarine antagonist pentiphan in the prevention of haloperidol-induced catalepsy in rats exceeds the activity of such traditional antiparkinsonian drugs as cyclodol and amedin. PMID- 9854622 TI - [The effect of gidazepam on the activity of the sympatho-adrenal system in operators working under the usual and complicated conditions]. AB - In healthy volunteers a single dose (0.05 g) of hydazepam does not change, but in subjects with psychogenic stress-reactions one-week course of the drug (0.05 g/24 h) intensifies to optimum activation of the sympathoadrenal system in response to standard operator load, inducing prevalent excretion of norepinephrine but not of epinephrine in the urine and maintains a high level of dopamine and DOPA excretion. In operators working in a state of hypercapnia and hyperthermia a single dose of hydazepam (0.05 g) blocks stress activation of the sympathoadrenal system. In a state of hyperthermia hydazepam normalizes the blood histamine and serotonin content. In all cases hydazepam improves the condition of the operators and the quality of their work, which allows it to be considered an effective and safe agent for the correction of stress-induced disorders in operators. PMID- 9854624 TI - [The pharmacological modulation of group behavior and biorhythms in the dynamics of a chronic experiment]. AB - Cyclic daily experiments were conducted using the standard method of free behavior of animals in an "open field" (Opto-Varimex, USA) with by the hour recording of locomotor parameters. The neuromodulating activity of 124 pharmacological agents was compared according to the size and duration of their stimulating or depressant effect. PMID- 9854623 TI - [New approaches in correcting disorders of higher integrative brain functions in the pharmacotherapy of arterial hypertension with propranolol]. AB - In experiments on rats it is found that propranolol in a dose of 2 mg/kg has a negative effect on higher integrative functions of the brain. Nootropics- pyracetam (200 mg/kg), AKF-94 (20 mg/kg), PIR-87-6-O (50 mg/kg) and actoprotector bemityl (40 mg/kg)--normalized orientative-trying behavior, memory, and emotionality of the animals. A possible mechanism of the drug action and prospects of their clinical application are discussed. PMID- 9854625 TI - [GABA reuptake by rat brain slices in hypokinesia and under the influence of cinnarizine and flunarizine]. AB - Studies showed that 15-day restriction of motor activity inhibits the reuptake of GABA by sections of the rat brain cortex and hypothalamus. In prolonged intraperitoneal injection of 10 mg/kg cinnarizine in hyperkinesia, further inhibition of this process in hypothalamic sections is encountered on the 15th days. Flunarizin (1 mg/kg) administered in a like manner has no significant effect on GABA reuptake. A course of GABA injections in a dose of 5 mg/kg improves in rats with restricted motor activity the local blood flow in the frontal lobe of the brain. PMID- 9854626 TI - [A nondepolarizing muscle relaxant with rapidly developing and short-term action]. AB - The pharmacological properties of the new nondepolarizing myorelaxant IEM-1213 and of its mixture with tercuronium were studied in experiments on anesthetized cats. Intravenous infusion of a blocking dose of IEM-1213 did not cause a change in the level of arterial pressure and blockade of the sympathetic ganglia but induced blockade of the heart muscarine receptors. The effect of IEM-1213 develops more rapidly and lasts for a shorter time than that of dithylin. Intravenous infusion of a mixture of IEM-1213 and tercuronium constituting 35 and 60% of the blocking dose of the former and, respectively, 35 and 20% of the blocking dose of the latter causes an effect similar in the time of its development to that of intravenous infusion of a total dose of IEM-1213 alone. PMID- 9854627 TI - [The anti-ulcer action of dibunol (Tonarol)]. AB - In experimental study of antiulcerative activity of dibunol on various models of gastric ulcers in rats the drug caused a marked antiulcerative effect in all of them, reduced the incidence of ulcer formation, and shortened the time of ulcer healing. In a model of "acetic" ulcer dibunol oil solution led to quick normalization of lipid peroxidation in the gastric mucosa, which was evidence of high antioxidant activity in cases of ulcer lesions. PMID- 9854628 TI - [The evaluation of the action of microbial enzyme preparations on the motility of the rat and canine small intestine]. AB - The effect of combined microbial enzyme agents trizyme and triaze containing alkaline and neutral proteases, lipase, and amilase on the evacuation capacity of the small intestine of conscious rats and of the dog small intestine operated on by the Thiry-Vella method. Therapeutic doses of the enzymatic agents had no noticeable effect on the evacuation function of the intestine. The contraction of bands of the rat intestine induced by electrical stimulation depended on the concentration of the perfusate of the solution of the enzymatic agents. Low concentration of the agents caused no effect or weakly activated the contractions of the smooth intestinal muscles. Doses of the agents higher than the mean therapeutic doses caused decrease of the contraction amplitude of the intestinal bands by 30-40%. PMID- 9854629 TI - [The effect of an extract of Baikal skullcap (Scutelleria baicalensis) on succinic acid oxidation by the brain mitochondria in rats with hypoxia]. AB - Oxidative phosphorylation in the mitochondria of rat brain in normobaric hypercapnic hypoxia and preventive administration of Scutellaria baicalensis extract was studied by polarography. Hypoxia causes diminution of energy of the rat brain mitochondria with separation of oxidative phosphorylation and inhibition of the reactions of rapid metabolic cluster of Krebs' cycle. Scutellaria baicalensis extract prevents diminution of energy of the brain mitochondria in hypoxia, inhibits restriction of succinate dependent energy production, and preserves intactness of the mitochondrial membranes. PMID- 9854630 TI - [The immunomodulating and antioxidants actions of plant preparations and abisib in disordered lipid metabolism]. AB - Five intraventricular injections of phytopreparations lohein and abisib in a dose of 0.2 or 0.5 ml stimulated dose-dependently the development of a T-dependent and T-independent immune responses in rats kept on the usual diet, and increased or normalized the development of these processes in rats kept on a atherogenic diet. The drugs lessened lipid peroxidation and the biochemical syndromes of hepatocyte affection in animals given an atherogenic diet. The immunomodulating effect of phytopreparations is mediated by cytokines of adherent and nonadherent splenic cells, the biochemical effects only by cytokines of adherent splenocytes. PMID- 9854631 TI - [The immunological effects of perftoran]. AB - The effect of perftocarbonic emulsion-perftoran on the course of experimental lipopolysaccharide intoxication was studied in albino mice. Resistance of the animals to the endotoxin reduced 3 days after administration of 10 ml/kg of the preparation and increased 6 days after its administration. The effect of perftoran on the dynamics of some parameters of immunity and nonspecific resistance in CBA mice and guinea pigs was studied in immunodeficiency states induced by injection of 5-fluorouracil, cyclophosphamide, and irradiation. Activation of the phagocytic system occurred 3 days later, and activation of the parameters of cellular and humoral immunity 6 days later. PMID- 9854632 TI - [A comparative study of the mechanisms of the immunosuppressive action of morphine and adrenaline in leukocytes in vitro]. AB - The effect of morphine (10(-6) M) and adrenaline (0.2 microgram/10(6) cells) on the oxidant and antioxidant activity in cells of the immunophagocytic system was compared in vitro. Morphine and adrenaline caused different changes in production of active forms of oxygen (AFA) and antioxidant activity of the enzymes superoxide dismutase and glutathione reductase in neutrophils, monocytes, and lymphocytes in human peripheral blood. The effect of these drugs, however, was unidirectional in character in each type of cells. Eleutherococcus (obtained in the drug-store and diluted 10- to 11-fold) or indomethacin (10(-7) M) injected together with morphine removed the morphine-induced increase of APA production in the neutrophils, they also prevented the effect of morphine on enzyme activation in the neutrophils and monocytes, but had no influence on the effect of morphine on the antioxidant enzymes in the neutrophils. The results of the study point to the existence of common mediators in direct action of adrenaline and morphine on the cells of the immunophagocytic system. PMID- 9854633 TI - [The pharmacokinetics and metabolism of heptapeptide--a prospective synthetic analog of tuftsin with psychostimulating action in rats]. AB - The pharmacokinetics of a new heptapeptide with psychotropic activity was studied in rats in experiments in vitro and in vivo by means of high-performance liquid chromatography. The heptapeptide was detected in its natural form in rat blood plasma for 7-10 min; its T1/2 exceeded that of other oligopeptides of similar structure. The structure of the main pharmacologically active heptapeptide metabolite was identified by high performance liquid chromatography; it proved to be tetrapeptide Thr-Lys-Pro-Arg (taphtsin). The heptapeptide was distributed in well vascularized organs of rats: liver > kidneys > heart. PMID- 9854634 TI - [New data on the pharmacokinetics of aminostigmine]. AB - The results of studying the pharmacokinetics of aminostigmine, a new reversible cholinesterase inhibitor produced in Russia, are discussed. Tissue radioactivity after intragastric administration of a toxic dose of aminostigmine was quite quickly absorbed from the intestinal lumen. The half-life period of aminostigmine exceeded considerably that of earlier studied carbamates and correlated with the clinical manifestations of the drug effects. This confirms that the use of aminostigmine in the treatment of neurologic and somatic diseases is preferable. PMID- 9854635 TI - [The pharmacokinetics of poliosm]. AB - Experiments were conducted on cats to study the pharmacokinetics of polyosm possessing diuretic and antiedemic properties whose primary acting component is polyethyleneoxide 400. Biexponential dependence of the blood drug concentration on the time with T1/2 values of 41 min and 4.8 h was revealed. In the first 5 h after intravenous infusion of the drug 60.8% of the introduced dose of polyethyleneoxide 400 (1 g/kg) was excreted through the kidneys. PMID- 9854636 TI - [The hepatoprotective action of ellagotannins]. AB - The hepatoprotective properties of altan (obtained on the basis of ellagotannines from the cones of black alder Alnus glutinosa) are studied on the model of acute damage to the liver with tetrachloromethane. It is found that altan exhibits the hepatoprotective activity even in a dose of 1 mg/kg which is tenfold smaller compared to the dose of traditional flavonoid-based drugs. Altan limits choleopoiesis disorder, has an antiinflammatory and membrane stabilizing effect, and recovers physiological antioxidant system. PMID- 9854637 TI - [The effect of sodium thiosulfate on nonspecific body resistance and on the immune reactions in acute acrylonitrile poisoning]. AB - Experiments on noninbred Wistar mice and rats showed that acute acrylonitrile (0.5 LD50) intoxication leads to decrease of nonspecific resistance of the organism and humoral and cellular immunity responses. The antidote sodium thiosulfate increases suppression of the responses under study. PMID- 9854638 TI - [The effect of polyamine on the structure of the free amino acid pool in the blood plasma and in the liver of animals with chronic alcoholic intoxication]. AB - The effect of polyamine on the total content of free amino acids and the structure of the amino acid fund in blood plasma and hepatic tissue was studied in rats with chronic alcoholic intoxication. Rapid utilization of exogenic introduced amino acids and a normalizing effect of the drug on the structure of the amino acid fund were demonstrated. The high concentration of aromatic amino acids in the drug may be among the causes of the increased content of this fraction of amino acids in the tissues studied after administration of the drug. PMID- 9854639 TI - [The effect of GABA-ergic agents on the electrolyte balance under hypokinesia]. AB - The effect of hypokinesia on the blood electrolyte balance was studied. In restricted motor activity marked deviations of the type of hypokalemia and hypercalcemia were encountered, which could promote the development of various pathological processes, including disorder of central and regional hemodynamics. GABA and piracetam had no effect on the blood serum electrolyte content in the early periods of hypokinesia. In later periods of hypokinesia both drugs under examination corrected the disorders of blood electrolyte balance induced by prolonged restriction of motor activity. PMID- 9854640 TI - [The comparative characteristics of the metabolic effects of amtizol and trimetazidine in acute hypoxia]. AB - Experiments were conducted on rats to study the effect of amtizol and trimetazidine on energy metabolism and lipid peroxidation in the brain, heart, and liver in acute hypoxia. Hypoxia was modelled by "raising" the animals in a pressure chamber to a "height" of 8,000-11,000 m, the exposure lasted 30 min. Equal doses (25 mg/kg) of the drugs were injected intraperitoneally 30 min before the "ascent". It was found that amtizol, just like trimetazidine, prevented energy metabolism disorders and activation of lipid peroxidation in the organs. The metabolic effects of the drugs were unidirectional, but the antioxidant activity of amtizol was pronounced more. PMID- 9854641 TI - [Arousal and the psychopharmacology of anxiety in experimental models]. AB - Locomotion is a behavioural marker of arousal in animals whereas exploration serves as a reliable criterion for anxiety level. Traditional views of central excitatory mechanisms consider them as pro-arousal and anxiogenic. Central inhibitory mechanisms are thought to be sedative and anxiolytic. However, data exist on paradoxical anti-anxiety effects of low doses of certain anxiolytics in a number of experimental models. Likewise, anxiolytics might exert paradoxical pro-arousal/anxiogenic profiles in animals. Both such data are difficult to interpret within traditional framework of arousal/anxiety concepts. The paper reviews data on complex non-linear interrelationships between anxiety and arousal and supports a theory of putative "anxiolytic" (comfort or pleasurable) emotions evoked by mild, but not high, arousal. PMID- 9854642 TI - [Cytokines and microsomal oxidation]. PMID- 9854643 TI - [Angiogenin-induced angiogenesis as a target in the therapy of tumors]. PMID- 9854644 TI - Doctors' diagnoses and patients' perceptions. Lessons from chronic fatigue syndrome. PMID- 9854645 TI - Fluvoxamine for somatoform disorders: an open trial. AB - Although the pharmacologic treatment of somatoform disorders has scarcely been investigated, there is reason to believe that antidepressants might be useful. We examined the response of 29 patients with somatoform disorders from a general medicine clinic to a selective serotonin reuptake inhibitor, fluvoxamine. The drug was administered in doses of up to 300 mg daily for 8 weeks. Sixty-one percent of the patients who took medication for at least 2 weeks were at least moderately improved. In addition to antidepressant effects, fluvoxamine had other beneficial effects and was well-tolerated. The benefits of drug therapy were modest but appear to warrant a placebo-controlled trial. PMID- 9854646 TI - Posttraumatic stress disorder in response to HIV infection. AB - This study investigated the psychological impact of HIV infection through assessment of posttraumatic stress disorder in response to HIV infection. Sixty one HIV-positive homosexual/bisexual men were assessed for posttraumatic stress disorder in response to HIV infection (PTSD-HIV) using a modified PTSD module of the DIS-III-R. Thirty percent met criteria for a syndrome of posttraumatic stress disorder in response to HIV diagnosis (PTSD-HIV). In over one-third of the PTSD cases, the disorder had an onset greater than 6 months after initial HIV infection diagnosis. PTSD-HIV was associated with other psychiatric diagnoses, particularly the development of first episodes of major depression after HIV infection diagnosis. PTSD-HIV was significantly associated with a pre-HIV history of PTSD from other causes, and other pre-HIV psychiatric disorders and neuroticism scores, indicating a similarity with findings in studies of PTSD from other causes. The findings from this preliminary study suggest that a PTSD response to HIV diagnosis has clinical validity and requires further investigation in this population and other medically ill groups. The results support the inclusion of the diagnosis of life-threatening illness as a traumatic incident that may lead to a posttraumatic stress disorder, which is consistent with the DSM-IV criteria. PMID- 9854647 TI - Spinal cord injury and psychological response. AB - Psychological adjustment and psychopathological morbidity issues during rehabilitation of patients with spinal cord injury, have been documented in international literature. However, most authors are faced with methodological difficulties, and results are contradictory. In this prospective study, the first to be made in the Portuguese population, a sample of 65 patients being treated in a rehabilitation unit during the years of 1993, 1994 and 1995, was obtained. The authors study the type of psychological response, when it does occur, which personality traits point to less suffering, which coping mechanisms are used by the better adjusted patients and the differences between the scores of paraplegic and quadriplegic patients. Two assessments were made. The following assessment instruments were used: an anamnestic data questionnaire, the SCL-90-R (Derogatis, 1983), the EPI (Eysenck & Eysenck, 1984), the Coping Styles Evaluation Scale (Figueira, 1990). The second assessments were carried out with the SCL-90-R only. The findings indicate that psychopathological scores consistent with depression occurred in 60% of patients if we consider any evaluation. Sleep disturbances, suicide ideation and guilt occurred in the same proportion. In 33% of them, we found persistent depressive scores in the two assessments. The authors find a highly significant positive correlation between psychopathology and neuroticism. On the contrary, the extroversion dimensions of EPI seem to be a good prognosis predictive factor as far as the occurrence of psychopathology is concerned. No differences in the psychopathological response were found concerning the paraplegic-quadriplegic situation. PMID- 9854648 TI - A survey of family physicians and psychiatrists. Psychotropic prescribing practices and educational needs. AB - The purpose of this study was to compare the psychotropic prescribing practices of family physicians and psychiatric physicians, and to assess their satisfaction with education about psychotropic prescribing. A survey was mailed to 461 active members of the Minnesota Psychiatric Society and to a randomized sample of 461 active members of the Minnesota Academy of Family Physicians. Major depression, panic disorder, and dementia with behavioral disturbance were selected for study. For each disorder, we asked for the estimated number of patients seen and treated as well as a list in order of preference of the three medications they most commonly use. Those surveyed were also asked about their level of satisfaction with the training they received in medical school, residency, and continuing medical education (CME) courses on the topic of psychotropic prescribing. The two physician groups reported similar prescribing practices for the single-drug treatment of depression. Family physicians more frequently prescribed minor tranquilizers and older generation psychotropics than did psychiatrists when treating panic disorder and dementia with behavioral disturbance. Patients with depression were the least likely to be referred to other health professionals. Neither physician group was satisfied with medical school education in this area; family physicians were less satisfied with residency training and in related CME courses than were psychiatric physicians. Additional study is needed to better understand interspecialty practice variation for commonly encountered psychiatric disorders. Medical education programs at all levels could better meet the reported needs of their graduates by providing more attention to prescribing psychotropic agents. PMID- 9854649 TI - Mnemonics for DSM-IV substance-related disorders. AB - A series of acronyms was developed to assist in recalling the DSM-IV diagnostic criteria for substance-related disorders. In order to facilitate recall, most of the mnemonics are related in some way to the disorder itself. In general, each letter of the acronym relates to an element of the diagnostic criteria. These mnemonics are intended to be helpful in a variety of academic and medical settings. PMID- 9854650 TI - Determinants of psychotropic drug usage in a general intensive care unit. AB - During a 3-month period, determinants of psychotropic drug utilization (sex, age, length of stay, reason for admission, disease severity) and data on psychotropic consumption (type of medication--antidepressants, benzodiazepines and antipsychotics--dosage and length of treatment) were retrospectively collected in a general intensive care unit of a Dutch university hospital. Daily exposure to psychotropics was standardized in number of Defined Daily Doses (DDD). Benzodiazepines were used by 35.8% of all patients (137) during their stay in the ICU whereas 17.5% of all patients used a neuroleptic agent. Antidepressants were hardly prescribed. High doses of benzodiazepines (9.9 DDDs) and low doses of antipsychotics (0.5 DDDs) were prescribed, which probably reflect the unusual nature of this critically ill group of patients compared with the reference group for DDD's. Clear patterns of determinants of psychotropic drug use in ICU patients were found and both benzodiazepines, antipsychotics and combined use of these agents could be associated with the determinants assessed. The time patterns we found in terms of length of stay give clues for further investigations in order to rationalize psychotropic drug use in the management of severely ill and complex patients. PMID- 9854651 TI - Risk factors for substance use disorders among inpatients with major affective disorders in Taiwan Chinese. AB - An assessment of potential risk factors for substance use disorders was performed in 49 patients with major affective disorders in a general hospital acute care unit in Taiwan. The major depression patients were noted to have a higher prevalence of substance use disorder than bipolar disorder patients (p = 0.011). Those patients with substance use disorders were noted to be significantly male predominated (p = 0.043), to have a later onset age of affective disorder (p = 0.009), and to have more visits to the emergency room in the recent one year (p = 0.009). The sedatives-hypnotics-anxiolytics was the most frequently used forms of drug abuse. The major depression patients had a significantly higher sedative hypnotics-anxiolytics use disorder rate than the bipolar disorder patients (p = 0.001). All patients with alcohol use disorder were noted to have other substance use disorders as well. Fifty six percent of those patients with substance use disorder were polysubstance users. Eighty three percent of the male patients with substance use disorder had poly-substance use disorder. PMID- 9854652 TI - Childhood sexual trauma as a potential factor for noncompliance with endoscopic procedures. PMID- 9854653 TI - Childhood abuse, personality disorder and chronic fatigue syndrome. PMID- 9854654 TI - Treatment with risperidone of an acute psychotic episode in a patient with AIDS. PMID- 9854655 TI - Show me the money (but first show me the data!). PMID- 9854656 TI - An institution's initial experience with port-access minimally invasive cardiac surgery. AB - OBJECTIVE: To assess the learning curve associated with Port-Access minimally invasive cardiac surgery. DESIGN: Retrospective. SETTING: Single university hospital. PARTICIPANTS: Initial 10 patients undergoing Port-Access minimally Invasive cardiac surgery. INTERVENTION: Minimally invasive cardiac surgery. MEASUREMENTS AND MAIN RESULTS: All 10 patients experienced an uneventful intraoperative and immediate postoperative course. Only one patient experienced postoperative cardiovascular morbidity, which was an episode of new-onset atrial fibrillation after mitral valve surgery that was successfully treated with pharmacologic therapy. Extubation times and postoperative discharge times were less than historic controls receiving the same anesthetic technique at the same institution. CONCLUSION: This institution's initial experience with 10 patients undergoing Port-Access minimally invasive cardiac surgery suggests an acceptable learning curve and decreased extubation and postoperative discharge times, which should translate into reduced health care costs. PMID- 9854657 TI - Detection of postoperative myocardial ischemia by bedside ST-segment analysis in coronary artery bypass graft patients. AB - OBJECTIVE: To determine if the ST-segment monitoring software of a bedside electrocardiograph (ECG) monitor would detect postoperative myocardial ischemia (POMI) as reliably as the clinical gold standard 12-lead ECG, and to compare the characteristics of ischemia thus detected with prior studies performed using Holter monitoring. DESIGN: Prospective study. SETTING: University hospital. PARTICIPANTS: One hundred patients who had undergone coronary artery bypass grafting (CABG). INTERVENTIONS: Continuous ST-segment trends in leads II and V5 were recorded using Hewlett-Packard Merlin monitors postbypass until discharge from the postsurgical unit, and printouts were analyzed for episodes of ischemia. Simultaneous 12-lead ECGs and monitor strips were recorded during ischemic episodes in 24 patients and were independently analyzed by two blinded cardiologists quantitatively for ST-segment values and qualitatively for an overall ischemia rating. MEASUREMENTS AND MAIN RESULTS: The ST-segment values directly measured by the cardiologists on the simultaneous 12-lead ECGs and those recorded by the monitor during ischemic episodes were found to be clinically comparable (bias, 0.1 mm for both leads; precision, 0.5 mm lead II, 0.9 mm lead V5). The sensitivity of the monitor compared with 12-lead ECGs for the detection of POMI was 73%. Ninety-eight episodes of significant ST deviation were identified in 39 patients. The characteristics of the ischemia detected in this study were similar to those reported in other studies performed using continuous Holter-type monitoring. CONCLUSION: Bedside monitoring of the ST segment in leads II and V5 using Hewlett-Packard Merlin monitors after CABG surgery is as accurate for the measurement of ST deviation in those leads as the clinical gold standard of a 12-lead ECG read by an experienced cardiologist. However, the 12-lead ECG will detect POMI more reliably than an automated two-lead bedside ST-segment analyzer because it allows evaluation of more leads and of ST-segment and T-wave morphology. Bedside ST-segment monitoring in this study confirmed the high incidence of ischemia after CABG surgery shown previously using Holter monitoring. PMID- 9854658 TI - Real-time intraoperative transesophageal echocardiography--how useful? Experience of 5,016 cases. AB - OBJECTIVE: To evaluate transesophageal echocardiography (TEE) as an intraoperative monitoring modality and to assess its safety, reliability, and overall utility in real-time use during cardiac surgery. DESIGN: Prospective, observational cohort study performed from January 1993 to June 1997. SETTING: Operating room of a tertiary care hospital for cardiology and cardiovascular surgery. PARTICIPANTS: Five thousand and sixteen adult patients with acquired heart disease, who underwent 1,356 valve procedures and 3,660 coronary artery bypass graftings (CABGs). INTERVENTIONS: All patients were monitored with radial artery and pulmonary artery catheters, along with continuous TEE monitoring with a multiplane transducer. MEASUREMENTS AND MAIN RESULTS: Prebypass imaging yielded unsuspected findings that either helped or modified the surgical plan in 158 of 1,356 valve procedures (11.65%) and in 993 of 3,660 CABGs (27.13%). There were 3,217 TEE-guided hemodynamic interventions in 944 patients (25.79%) in the CABG group and 629 in 142 patients (10.47%) in the valve group. TEE was the sole guiding factor in initiating therapy in 23.53% of events, whereas it was supportive to other monitoring modalities in 76.46% of events. Postbypass TEE identified the need for graft revision in 29 patients (0.8%), intra-aortic balloon pump (IABP) requirement in 29 patients (0.8%), and inadequate valve repair in 28 patients (2.08%). For the entire series, 38.78% of patients benefited from prebypass and 39.16% from postbypass use of TEE. There were no complications attributable to the use of TEE in the entire series. There was 87% concordance between online interpretation by a trained anesthesiologist and offline analysis by a cardiologist. CONCLUSION: Intraoperative TEE is useful in formulating the surgical plan, guiding various hemodynamic interventions, and assessing the immediate results of surgery. It is safe and the results are reliable in the hands of trained anesthesiologists. PMID- 9854659 TI - Fibrinolysis in pediatric patients undergoing cardiopulmonary bypass. AB - OBJECTIVE: Thromboelastographic evaluation of the influence of fibrinolysis on blood loss and blood product transfusions in children during cardiac surgery. DESIGN: Prospective study. SETTING: University-affiliated, pediatric medical center. PARTICIPANTS: Two hundred seventy-eight consecutive children undergoing cardiac surgery. INTERVENTIONS: Blood sampling for coagulation tests, including native and protamine-modified thromboelastography. MEASUREMENTS AND MAIN RESULTS: Blood coagulation tests were measured before, during, and after cardiopulmonary bypass (CPB). Demographic data, perioperative blood loss, and blood product transfusions were prospectively recorded. Fibrinolysis was defined as thromboelastography of A30/MA less than 0.85 (MA, maximum amplitude; A30, amplitude 30 minutes after MA) and was noted in 3% of children pre-CPB, 16% during CPB, and 3% post-CPB. Fibrinolysis before CPB was associated with poor cardiac output. Fibrinolysis during CPB occurred in young children (aged 350 +/- 836 days) undergoing complex surgery with prolonged CPB (119 +/- 48.8 minutes) and deep hypothermia (25.6 degrees C +/- 4.7 degrees C). These patients received blood products after CPB and were not fibrinolytic after transfusion. They incurred similar blood loss (in mL/kg) and received similar volumes of blood products (mL/kg) as age-matched and surgery-matched patients without fibrinolysis. CONCLUSION: A group of children at risk for fibrinolysis during CPB was identified. However, fibrinolysis during CPB did not influence blood loss or the total volume of blood products transfused. PMID- 9854660 TI - The effect of three different doses of tranexamic acid on blood loss after cardiac surgery with mild systemic hypothermia (32 degrees C). AB - OBJECTIVE: Prophylactic administration of tranexamic acid (TA), an antifibrinolytic agent, decreases bleeding after cardiac surgery with systemic hypothermia (25 degrees C to 29 degrees C). Warmer systemic temperatures during cardiopulmonary bypass (CPB) may reduce bleeding and thus alter the requirement for TA. The effect of three different doses of TA on bleeding after cardiac surgery with mild systemic hypothermia (32 degrees C) is evaluated. DESIGN: Double-blind, prospective, randomized study. SETTING: University hospital. PARTICIPANTS: One hundred fifty adult patients undergoing aortocoronary bypass or valvular cardiac surgery. INTERVENTIONS: Patients received TA, 50 (n = 50), 100 (n = 50), or 150 (n = 50) mg/kg intravenously before CPB with mild systemic hypothermia. MEASUREMENTS AND MAIN RESULTS: Blood loss through chest drains over 6, 12, and 24 hours after surgery and total hemoglobin loss were measured. Autotransfused blood, transfused banked blood and blood products, and coagulation profiles were measured. Analysis of variance on log-transformed data for blood loss and confidence intervals (CIs) of 0.95 were calculated and transformed to milliliters of blood. No patient was re-explored for bleeding. Blood loss at 6 hours was statistically greater in the 50-mg/kg group compared with the other two groups (p = 0.03; p = 0.02). Total hemoglobin loss was statistically greater in the 50-mg/kg group compared with the 150-mg/kg group (p = 0.04). There was no statistical difference in blood tranfusion rate or coagulation profiles among the three groups. However, preoperative hemoglobin level was statistically lower in the 150-mg/kg group compared with the other two groups (p = 0.01). CONCLUSION: Of the three doses of TA studied, the most efficacious and cost-effective dose to reduce bleeding after cardiac surgery with mild hypothermic systemic perfusion is 100 mg/kg. PMID- 9854662 TI - Pain management in cardiac surgery patients: comparison between standard therapy and patient-controlled analgesia regimen. AB - OBJECTIVE: To compare standard nurse-based pain therapy with a patient-controlled analgesia (PCA) regimen. DESIGN: Prospective, randomized study. SETTING: Single institutional, clinical investigation in an urban, university-affiliated hospital. PARTICIPANTS: Sixty patients undergoing elective first-time cardiac surgery were included. INTERVENTIONS: In 30 patients, a standard analgesic regimen was used, and in 30 patients, a PCA regimen was used. The perioperative and postoperative management was similar for all patients. MEASUREMENTS AND MAIN RESULTS: Degree of sedation, satisfaction, and pain (by visual analog scale [VAS]) was assessed within the first 3 postoperative days. Vital capacity (VC) and forced expiratory volume in 1 second (FEV1) were measured using a portable spirometry system. Cortisol and troponin T (TnT) plasma levels were also measured. The expectation of pain was similar in both groups, and the postoperative pain score was significantly lower in the PCA than in the standard group throughout the study period. Significantly more piritramid was used in the PCA (total, 75.6 +/- 33.4 mg) than in the standard group (total, 20.1 +/- 31.9 mg). VC and FEV1 were significantly lower in the standard group compared with the PCA patients. Cortisol and TnT plasma levels were similar in both groups. Frequency of side effects were similar for both groups. CONCLUSION: Because of the beneficial effects with regard to degree of pain and satisfaction, pain management using PCA systems can be recommended for cardiac surgery patients. It appears to be superior to standard nurse-based pain therapy. PMID- 9854661 TI - Dopexamine unloads the impaired right ventricle better than iloprost, a prostacyclin analog, after coronary artery surgery. AB - OBJECTIVE: To evaluate the ventricle-unloading properties of dopexamine and iloprost and to compare their effects on right ventricular (RV) function and oxygen transport in patients with low RV ejection fraction (RVEF) after cardiac surgery. DESIGN: A prospective, randomized, double-blind, cross-over, clinical study. SETTING: University hospital. PARTICIPANTS: Twenty patients with proximal total stenosis of the right coronary artery studied immediately after coronary artery surgery. INTERVENTIONS: Treatment drugs were administered in a random order in doses equipotent with respect to cardiac output response. Infusion rates were increased stepwise to induce a 25% increase in cardiac index. A washout period of 60 minutes was allowed between treatments. MEASUREMENTS AND MAIN RESULTS: Central hemodynamics, RV function assessed by the EF (fast-response thermodilution), end-systolic and end-diastolic volumes, and systemic oxygenation were measured before and after the first drug, after the washout period, and after the second drug. Central filling pressures remained constant during treatments. Both drugs decreased pulmonary vascular resistance index, but iloprost was more effective (p < 0.05). Iloprost decreased mean arterial and pulmonary artery pressure, which were unaffected by dopexamine. Dopexamine increased EF significantly more than iloprost (p < 0.001). End-systolic volume index decreased subsequent to dopexamine only (p < 0.001). Iloprost increased intrapulmonary shunt more than dopexamine (p < 0.001). Changes in oxygen delivery, consumption, and extraction were similar. CONCLUSION: The findings suggest that dopexamine is more effective than iloprost for support and unloading of the postoperatively disturbed RV in terms of RVEF and end-systolic volume. The reduction of pulmonary vascular resistance after administration of iloprost without a decrease in end-systolic volume might not be considered a reduction of RV afterload. Iloprost increases the pulmonary shunt fraction, however, more than dopexamine, indicating a more prominent vasodilator effect. PMID- 9854663 TI - Incidence of arrhythmias after thoracic surgery: thoracotomy versus video assisted thoracoscopy. AB - PURPOSE: Atrial arrhythmias, especially supraventricular tachycardia (SVT) and atrial fibrillation, are common after thoracotomy and lung surgery. There are few existing data on the incidence of postoperative arrhythmias after video-assisted thoracoscopy (VAT). The purpose of the present investigation was to retrospectively determine the incidence of postoperative arrhythmias in patients who underwent VAT compared with those who underwent thoracotomy, and which factors are associated with an increased risk for arrhythmias in both groups. DESIGN: A retrospective investigation. SETTING: A metropolitan university hospital. PARTICIPANTS: The medical records of 124 patients who underwent thoracotomy and 81 patients who underwent VAT over a 2-year period were reviewed. MEASUREMENTS AND MAIN RESULTS: There was a 17% incidence of atrial arrhythmias after thoracotomy and 10% after VAT, but the difference was not statistically significant. In both groups, atrial fibrillation was the most common atrial arrhythmia. CONCLUSION: Patients receiving digoxin were at higher risk for postoperative arrhythmias. Patients older than 65 years were at risk for arrhythmias after thoracotomy and patients older than 80 years were at risk for arrhythmias after VAT. Patients who had postoperative arrhythmias had prolonged hospital stays compared with patients who did not have arrhythmias. PMID- 9854665 TI - Minimally invasive direct coronary artery bypass procedure using a high thoracic epidural plus general anesthetic technique. PMID- 9854664 TI - Thoracic epidural anesthesia does not affect functional recovery from myocardial stunning in sevoflurane-anesthetized dogs. AB - OBJECTIVE: A beneficial effect of thoracic epidural anesthesia (TEA) on recovery from myocardial stunning was previously shown in awake dogs. The aim of this study was to investigate the effects of TEA on recovery from myocardial stunning in sevoflurane-anesthetized dogs. DESIGN: Randomized animal study. SETTING: Animal laboratory of a university hospital. PARTICIPANTS: Chronically instrumented mongrel dogs. INTERVENTIONS: Six dogs were chronically instrumented for measurement of hemodynamics and myocardial wall thickening fraction (WTF). The following experiments were performed on separate days in a crossover fashion: (1) 10 minutes of ischemia of the left anterior descending (LAD) coronary artery during sevoflurane anesthesia without TEA and (2) 10 minutes of ischemia during sevoflurane anesthesia with TEA. MEASUREMENTS AND MAIN RESULTS: WTF was measured awake (baseline) and at predetermined time points until complete recovery of myocardial function occurred. Induction of anesthesia led to a decrease of WTF compared with baseline. Induction of ischemia led to a further decrease of WTF to negative values, which returned to positive values within the first minute of reperfusion. There were no differences between the two experimental conditions at any of the time points measured. In awake dogs, TEA improved the recovery from myocardial stunning compared with the control experiment. There was no difference between conscious dogs with TEA or sevoflurane-anesthetized dogs with or without TEA. CONCLUSION: TEA has no additional protective effect on the recovery of WTF during sevoflurane anesthesia. PMID- 9854666 TI - Pericardial effusion--a life saver! PMID- 9854667 TI - Biventricular failure after pericardial window. PMID- 9854668 TI - The electrocardiogram complements a chest radiograph for the early detection of pneumothorax in post-coronary artery bypass grafting patients. PMID- 9854669 TI - Accurate detection of pulmonary embolism using epicardial echocardiography during right nephrectomy in a patient with renal cell carcinoma. PMID- 9854670 TI - Transesophageal echocardiography during organ transplantation. PMID- 9854671 TI - The use of multimedia to teach new cardiac anesthesia procedures. AB - Cardiac anesthesia is a complex field requiring extensive training and experience. Multimedia can be used effectively to teach new, complex, and rapidly developing anesthetic and surgical techniques. Minimally invasive Port-Access cardiac surgery is an area in which the computer can be used for educational purposes. As cardiac anesthesiologists become more adept at developing computerized instructional methods, and as new techniques in medicine emerge, greater use can be made of the computer as a valuable teaching tool. PMID- 9854672 TI - Case 6--1998. Perioperative anesthetic management of a patient presenting for resection of a tracheal mass. PMID- 9854673 TI - Pro: heparin-bonded circuits represent a desirable option for cardiopulmonary bypass. PMID- 9854674 TI - Con: heparin-bonded cardiopulmonary bypass circuits do not represent a desirable and cost-effective advance in cardiopulmonary bypass technology. PMID- 9854675 TI - An unexpected intracardiac mass. PMID- 9854676 TI - Endocoronary sinus catheter--Indian experience. PMID- 9854677 TI - Platelet concentrate effects on thromboelastography. PMID- 9854678 TI - The venous ACT after heparin--how much discard? PMID- 9854679 TI - Inadvertent closure of the superior vena cava after decannulation: a potentially catastrophic complication after termination of bypass. PMID- 9854680 TI - The modulation of glucocorticoid receptor content by 3-O-methyl-D-glucose transport in human mononuclear leukocyte in obesity. AB - Glucocorticoid receptors (GR) and 3-O-methyl-D glucose (3-O-MG) transport were determined in mononuclear leukocytes (MNL) from 11 abdominal obese subjects, 10 pituitary-dependent Cushing's syndrome (Cushing's disease) and 10 healthy controls. Using a whole-cell competitive binding assay and 3H-dexamethasone as tracer, MNL of abdominal obese subjects were found to have 4855 +/- 1389 sites/cell which was significantly lower (p < 0.05) than controls (6234 +/- 1568 sites/cell), although no significant difference was found in the mean serum cortisol level. Their mean Kd (affinity) was also significantly lower than that found in the healthy controls (obese Kd:2.92 +/- 0.84 nmol/l, control Kd: 4.55 +/ 0.67 nM, p < 0.05). On the other hand, the receptor characteristics in Cushing's disease patients were within the normal range. At the same time, 3-O-MG transport was determined in the same subjects. In Cushing's disease, 3-O-MG transport was within the normal range, whereas in abdominal obesity this value was significantly lower than the healthy controls (abdominal obese: 31.90 +/- 8.20; control: 46.26 +/- 12.91 fmol/10(6) cell, min, p < 0.05). We also found a positive correlation between 3-O-MG transport and GR binding capacity in abdominal subjects (r = 0.89, p < 0.001), however we did not find such a correlation in Cushing's disease (r = 0.60, p > 0.05). These results indicated that, in abdominal obesity, the GR binding capacity in MNL is influenced by the changes in glucose transport. PMID- 9854681 TI - Outcome of pregnancy after radioactive iodine treatment for well differentiated thyroid carcinomas. AB - This study sought to determine the outcome of pregnancy in female patients with differentiated thyroid carcinoma who became pregnant after radioactive iodide treatment. A total of 779 female thyroid cancer patients were treated at Chang Gung Medical Center in Linkou between January 1977 and December 1995. The medical records of these patients were reviewed retrospectively. Thirty-seven of these patients had well differentiated thyroid carcinoma receiving 131I treatment and conceived at a mean age of 27.97 +/- 3.49 year-old. A total of 58 pregnancy episodes were recorded during this study period. Among these 37 patients, 3 episodes of artificial abortion, 8 episodes of spontaneous abortion and 2 threatened abortions were observed. These patients delivered a total of 47 babies including 3 premature babies. Seven of these patients conceived within 6 months after the last administration of 131I, including 2 cases within 1 month, 4 cases within 4 months, and 1 patient within 5 months. Of these 7 patients, only one patient who conceived within 6 months after the last administration of 131I (14.3%) had a spontaneous abortion. The present results suggest that previous administration of 131I in female patients with well differentiated thyroid cancer does not result in demonstrable adverse effects in subsequent pregnancies. However, further studies involving long-term follow-up of children delivered by mothers who became pregnant within 6 months after the last administration of 131I is needed to further elucidate the possible chronic effects and sequelae of 131I therapy on subsequent pregnancies. PMID- 9854682 TI - Angiotensin II stimulates both aldosterone secretion and DNA synthesis via type 1 but not type 2 receptors in bovine adrenocortical cells. AB - Angiotensin II (Ang II) type 2 receptor (AT2) has been shown to counteract the type 1 receptor (AT1)-mediated biological actions of Ang II in the cardiovascular system. The biological significance of AT2 receptor in the adrenals however remains unknown. In the present study, we investigated the roles of AT1 and AT2 receptor subtypes in the regulation of aldosterone secretion and DNA synthesis in bovine adrenocortical zona glomerulosa cells in vitro. Ang II (1 mumol/l) stimulated aldosterone secretion was completely suppressed by AT1 antagonist CV 11974 but not affected by AT2 receptor antagonist PD-123319. Effects on DNA synthesis were investigated by determining the incorporation of BrdU into the nuclei of the cultured zona glomerulosa cells. Ang II (1 mumol/l)-stimulated DNA synthesis of the cells was also completely suppressed by CV-11974 but not by PD 123319. These results suggest that AT1 receptor but not AT2 receptor is the predominant receptor subtype which mediates the Ang II-stimulated aldosterone secretion and cell growth in bovine adrenocortical cells. PMID- 9854683 TI - Influence of galanin and serotonin on the endocrine response to Hexarelin, a synthetic peptidyl GH-secretagogue, in normal women. AB - Hexarelin (HEX) is a synthetic GH-secretagogue (GHS) which acts on specific receptors either at the pituitary or the hypothalamic level to stimulate GH release both in animal and in man. Like other GHS, HEX possesses also PRL-, ACTH- and cortisol (F)-releasing activity but the mechanisms underlying these effects are even less clear. On the other hand, galanin (GAL) and serotonin play an important role in the neural control of GH, PRL and ACTH secretion both in animal and in man. In order to study the interaction between HEX and GAL and to verify whether serotoninergic mechanisms underly the endocrine effects of GHS, in 12 normal young volunteers (24-30 yr) the following tests were performed: group A (N = 5), HEX (2.0 micrograms/kg i.v. at 0 min), GAL (15.0 micrograms/kg i.v. from 0 to 60 min) and HEX + GAL; group B (N = 7), HEX alone and preceeded by cyproeptadine (CYPRO, 8 mg os at -60 min). In group A, the GH response to HEX (1204.2 +/- 312.9 micrograms*min/L) was higher (p < 0.05) than that to GAL alone (305.6 +/- 35.5 micrograms*min/L) and was not modified by GAL co-administration (1021.8 +/- 249.9 micrograms*min/L). PRL secretion was increased to the same extent by HEX and GAL (507.9 +/- 81.1 and 743.0 +/- 164.7 micrograms*min/L) which showed no interaction (603.5 +/- 75.7 micrograms*min/L). HEX elicited an increase in both ACTH and F secretion (924.5 +/- 169.7 pg*min/ml and 6131.3 +/- 616.6 micrograms*min/L) while GAL had no effect when given alone (759.5 +/- 185.5 pg*min/ml and 5350.3 +/- 755.6 micrograms*min/L) and did not modify the effect of HEX (891.3 +/- 159.2 pg*min/ml and 5877.8 +/- 554.4 micrograms*min/L). In group B, the GH response to HEX (1636.4 +/- 267.5 micrograms*min/L) was blunted by CYPRO (1164.8 +/- 212.3 micrograms*min/L) but this difference did not attained statistical significance. On the other hand, CYPRO did not modify the HEX-induced PRL (599.5 +/- 129.2 vs 638.9 +/- 131.9 micrograms*min/L), ACTH (1282.8 +/- 222.0 vs 1330.2 +/- 347.0 pg*min/ml) and F response (4738.3 +/- 355.3 vs 4580.9 +/- 857.3 micrograms*min/L). Our present data demonstrate that Hexarelin has no interaction with galanin; thus thereotically, the stimulatory effect of GHS on GH and PRL secretion could involve, at least partially, a galanin-mediated mechanism. On the other hand, our data demonstrate that serotonin does not mediate the stimulatory effect of GHS on PRL, ACTH and cortisol; the intrinsic anticholinergic property of cyproeptadine could account for the trend toward its blunting effect on the GH response to Hexarelin. PMID- 9854684 TI - Thyroid follicular oncogenesis in iodine-deficient and iodine-sufficient areas: search for alterations of the ras, met and bFGF oncogenes and of the Rb anti oncogene. AB - To gain insights into the role of iodine deficiency in favoring thyroid tumorigenesis (particularly of the follicular histotype), 22 Sicilian patients with thyroid tumors were selected for having lived permanently in either one of two areas of different iodine availability. Eleven patients (age 46.1 +/- 14.6 years, mean +/- SD; 10 females and 1 male) were from the iodine-deficient (ID) areas of the provinces of Messina and Catania (mean urinary excretion of iodine = 48.1 micrograms/24 hours). Thyroid tumors were follicular or Hurthle cell adenomas (no. = 3), follicular carcinomas (FC, no. = 4), papillary carcinomas (PC, no. = 2) and anaplastic carcinomas (no. = 2). Eleven patients (age 47.1 +/- 15.2 years; 10 females and 1 male) were from the metropolitan area of Messina, an area of relative iodine-sufficiency (IS) (urinary excretion of iodine = 95.2 micrograms/24 hours). These 11 patients had serum levels of TSH that were significantly lower than the corresponding values of the 11 patients from the ID area (0.76 +/- 0.33 vs 1.80 +/- 1.22 mU/l, p = 0.01) The tumors of the 11 patients from the IS area were: follicular or Hurthle cell adenomas (no. = 6), Hurthle cell carcinoma (no. = 1), FC (no. = 2), PC (no. = 2). Molecular biology studies revealed that both the normal as well as the tumor tissue of all 22 patients did not harbor any of the three classical activating mutations (codons 12, 13 and 61) in any of the three ras oncogenes. Similar negative results were obtained as far as loss of heterozygosity of the retinoblastoma (Rb) anti oncogene is concerned. Immunohistochemistry studies were performed to investigate expression of c-met and basic fibroblast growth factor (bFGF) proto-oncogenes. Only one Hurthle cell carcinoma and the two PC from the IS group, and one FC and the two PC from the ID group stained for the c-met oncogene. Expression of c-met was greater (3+) in the four PC (concerning 70-80% of the tumor cells) than in the other two cancers (1+; < 5% of the tumor cells). In the IS group, positivity for bFGF was detected in 3/6 adenomas, 1/2 FC, the Hurthle cell carcinoma and the two PC. In the ID group, positivity for bFGF was observed in 2/3 adenomas, 2/4 FC, the two PC and the two anaplastic carcinomas. The 8 positive cases from the ID group had a greater level of bFGF expression than the 7 positive cases from the IS group (intensity of staining = 2.0+ vs 1.57+). Interestingly, the greatest expression of bFGF was seen in the cases with peri-tumoral lymphocytic infiltration from either group. In the ID group correlations between (i.) pre intervention serum TSH and intensity of tumoral staining for bFGF, (ii.) serum TSH and per cent of tumoral cells reactive with anti-bFGF and (iii.) between intensity of staining for bFGF and per cent of tumoral cells bFGF +ve were higher than in the IS group. We conclude that activating mutations of ras, loss of DNA from the Rb locus and over-expression of both c-met and bFGF are of no pathogenetic relevance in driving thyroid tumorigenesis of iodine-deficient areas. PMID- 9854685 TI - Bone mineral density and circulating cytokines in patients with acromegaly. AB - Acromegalic patients present an increase of osteoblastic and osteoclastic activity, showing a different effect on the axial and appendicular skeletal structures. At this regard controversial data about bone mineral density (BMD) have been published in literature. In fact an increase of BMD levels in femoral neck and Ward's triangle without any difference in lumbar spine has been described. On the other hand normal BMD levels at forearm and reduced BMD levels at lumbar spine were found. These patients seem to have a reduction of trabecular BMD similar to postmenopausal osteoporotic patients despite normal or slightly elevated cortical BMD. Recently, it has been described that cytokines, in particular tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1), are implicated in the pathogenetic mechanism of postmenopausal osteoporosis. Taking into account that growth hormone (GH) can increase TNF-alpha and IL-1 secretion by mononuclear blood cells, the evaluation of possible relationship between the reduced BMD at lumbar spine and circulating cytokines levels was carried out in acromegalic patients. In addition we evaluated the effect of acute octreotide administration on serum TNF-alpha and IL-I concentrations. Eleven patients with active acromegaly and eleven healthy age-, sex-, weight- and heightmatched subjects were enrolled in this study. BMD was significantly reduced at lumbar spine (0.80 +/- 0.29 g/cm2 vs 1.02 +/- 0.11 g/cm2; p < 0.01), but not at femoral neck level or at Ward's triangle level (0.92 + 0.15 g/cm2 vs 0.97 + 0.11 g/cm2, p = NS; and 0.74 +/- 0.16 g/cm2 vs 0.85 +/- 0.1 g/cm2, p = NS) when compared to controls. Baseline serum levels of TNF-alpha and IL-1 were in the normal range both in patients and controls. After acute octreotide administration, no differences in circulating TNF-alpha and IL-1 levels were found. In conclusion, acromegalic patients present a reduced BMD at lumbar spine but not at femoral neck level and Ward's triangle. Circulating cytokines such as TNF-alpha and IL-1 are in the normal range. These data suggest that cytokines are not involved in the pathogenesis of GH-excess induced osteoporosis. The possibility that the GH excess might affect bone turnover inducing an increase of cytokines acting by a paracrine/autocrine mechanism cannot be ruled out. PMID- 9854686 TI - Effect of finasteride in idiopathic hirsutism. AB - Increased 5 alpha-reductase activity has been found in hair follicles of hirsute women, suggesting a pathogenetic role. The aim of the present study was to evaluate the effect of finasteride in the treatment of idiopathic hirsutism. Twenty-seven women with idiopathic hirsutism, aged 16-35 years, were treated for 6 months with finasteride, 5 mg once daily. Fourteen patients were on finasteride alone (group A), while the remaining received in addition an oral contraceptive (group B). Clinical, hormonal and biochemical evaluation were performed before, and after 3 and 6 months of treatment. Clinical evaluation was repeated 6 months after drug discontinuation in seven patients. Treatment was well tolerated by all patients; no side effects or adverse reactions were reported. A significant improvement of hirsutism was obtained by finasteride; clinical score observed at the 6th month of therapy was reduced from 11.71 +/- 2.23 to 7.92 +/- 1.81 (p < 0.05) and from 14.92 +/- 6.13 to 9.3 +/- 2.75 (p < 0.05) in group A and B, respectively. Clinical score in seven patients was still 8.61 +/- 2.28 (p < 0.05) 6 months after the end of therapy. Finasteride treatment alone (group A) induced a slight increase, though not significant, in serum androgens; DHT and SHBG did not change. In group B (finasteride plus oral contraceptive) total testosterone and free testosterone showed no significant decrease; after 6 months of therapy DHT was reduced significantly, while SHBG levels were increased. These data demonstrate that 5 alpha-reductase inhibition may be an effective treatment in women suffering from idiopathic hirsutism. This approach may be attractive due to the absence of adverse reactions, although the necessity of an adequate contraception should be kept in mind. PMID- 9854687 TI - Transient neonatal hypothyroidism after gestational exposure to amiodarone: a follow-up of two cases. AB - Amiodarone (AMD) is an antiarrhythmic drug which contains 37% of iodine. It can reach the fetus by transplacental passage and induce fetal hypothyroidism. Since in some pregnant women AMD represents a cardinal therapeutic opportunity, it is necessary to establish not only the risk of teratogenicity linked to fetal AMD exposure but also to evaluate the psychomotor development of children with neonatal thyroid dysfunction related to fetal AMD exposure. We report on two cases involving children with an AMD gestational exposure and transient neonatal hypothyroidism, who were followed-up until the age of 4 years 8 months and 5 years 6 months, respectively. Denver's developmental milestone test and Whechsler Preschool and Primary Scale of Intelligence (WPPSI) were administered to the patients in accordance to their age. A normal psychomotor development was observed in both patients with full scale IQ score, verbal and performance IQ scores within normal range. In conclusion, if these data were validated by larger studies, it might not be obligatory to discontinue AMD administration in cardiopathic pregnant women, since mental impairment may not necessarily occur in children with transient neonatal hypothyroidism caused by fetal AMD exposure. However, the evaluation of the thyroid function of these children is imperative. PMID- 9854688 TI - A thyrotropin-secreting pituitary adenoma as a cause of thyrotoxic periodic paralysis. AB - We describe a patient with thyrotoxic periodic paralysis (TPP) caused by a thyrotropin-secreting pituitary adenoma. The diagnosis TPP was based on the combination of episodes of reversible hypokalaemic paralysis, hyperthyroidism and electrophysiological findings. A thyrotropin-secreting pituitary adenoma was diagnosed on the basis of endocrinological function tests and MRI of the pituitary gland. Before transsphenoidal resection of the adenoma, treatment with octreotide restored euthyroidism both clinically and biochemically. Immunocytochemistry of the pituitary adenoma was positive for TSH exclusively. Incubation with octreotide or quinagolide induced decreased TSH and alpha-subunit production by the cultured adenoma cells, in agreement with the pre-operative in vivo data. This paper is the first to describe in vivo and in vitro characteristics of a thyrotropin-secreting pituitary adenoma in a patient presenting with periodic paralysis. PMID- 9854690 TI - Fetal goiter in a iodine-deficient area. PMID- 9854689 TI - Non-classic adrenal hyperplasia in hyperandrogenism: a reappraisal. PMID- 9854691 TI - Preparation of school nurse practitioners. PMID- 9854692 TI - Youth Risk Behavior Surveillance--United States, 1997. State and Local YRBSS Coordinators. AB - Priority health-risk behaviors, which contribute to the leading causes of mortality and morbidity among youth and adults, often are established during youth, extend into adulthood, and are interrelated. The Youth Risk Behavior Surveillance System (YRBSS) monitors six categories of priority health-risk behaviors among youth and young adults--behaviors that contribute to unintentional and intentional injuries; tobacco use; alcohol and other drug use; sexual behaviors that contribute to unintended pregnancy and sexually transmitted diseases (STDs) (including human immunodeficiency virus [HIV] infection); unhealthy dietary behaviors; and physical inactivity. The YRBSS includes a national school-based survey conducted by the Centers for Disease Control and Prevention as well as state, territorial, and local school-based surveys conducted by education and health agencies. This report summarizes results from the national survey, 33 state surveys, 3 territorial surveys, and 17 local surveys conducted among high school students from February through May 1997. In the United States, 73% of all deaths among youth and young adults 10-24 years of age result from only four causes: motor vehicle crashes, other unintentional injuries, homicide, and suicide. Results from the national 1997 YRBSS demonstrate that many high school students engage in behaviors that increase their likelihood of death from these four causes--19.3% had rarely or never worn a seat belt; during the 30 days preceding the survey, 36.6% had ridden with a driver who had been drinking alcohol; 18.3% had carried a weapon during the 30 days preceding the survey; 50.8% had drunk alcohol during the 30 days preceding the survey; 26.2% had used marijuana during the 30 days preceding the survey; and 7.7% had attempted suicide during the 12 months preceding the survey. Substantial morbidity among school-age youth, young adults, and their children also result from unintended pregnancies and STDs, including HIV infection. YRBSS results indicate that in 1997, 48.4% of high school students had ever had sexual intercourse; 43.2% of sexually active students had not used a condom at last sexual intercourse; and 2.1% had ever injected an illegal drug. Of all deaths and substantial morbidity among adults greater than or equal to 25 years of age, 67% result from two causes--cardiovascular disease and cancer. Most of the risk behaviors associated with these causes of death are initiated during adolescence. In 1997, 36.4% of high school students had smoked cigarettes during the 30 days preceding the survey; 70.7% had not eaten five or more servings of fruits and vegetables during the day preceding the survey; and 72.6% had not attended physical education class daily. These YRBSS data are already being used by health and education officials to improve national, state, and local policies and programs to reduce risks associated with the leading causes of morbidity and mortality. YRBSS data also are being used to measure progress toward achieving 21 national health objectives and one of the eight National Education Goals. PMID- 9854693 TI - Sexual coercion content in 21 sexuality education curricula. AB - Sexual coercion, a topic of relevance to school health personnel, may be as common in high school populations as in university populations. Twenty-one sexuality education curricula were examined for information on the topics of date rape, stranger rape, pressure, incest, sexual harassment, unwanted/inappropriate touch, and exploitation/victimization. Curricula scoring highest in total coverage also were the most comprehensive with six of the seven sexual coercion topics covered. Overall, pressure and exploitation/victimization received the greatest attention, while sexual harassment was not covered in any of the curricula. Common themes occurring within the coercion topic areas included guilt, communication/assertiveness skills, blame, drug use, premeditation, fear, sources of help. Results suggest sexuality education curricula have not responded to the increased concern regarding sexual harassment in schools. PMID- 9854694 TI - The association between smoking and unhealthy behaviors among a national sample of Mexican-American adolescents. AB - This study examined the relationship between smoking and participation in unhealthy behaviors among Mexican-American adolescents through a secondary analysis of national data. Mexican-American adolescents (N = 580), ages 10 through 18 years who were interviewed as part of the 1993 Teenage Attitudes and Practices Survey (TAPS II), were selected for analysis. Data collected included smoking status of the adolescent and participation in certain unhealthy behaviors. Among girls in the study, smokers were more likely to not wear a seat belt, be involved in physical fighting, not be involved in organized sports, perform poorly in school, say they like to do risky things, and ride in a car with a drunk or high driver. For boys, smoking was significantly associated with liking to do risky things, fighting, not attending church, and poor academic performance. These results suggest that Mexican-American adolescents who smoke may be at higher risk for engaging in behaviors that could compromise their health and safety, and for not being involved in activities that may exert a protective influence. PMID- 9854695 TI - Promoting holistic child development: a collaborative school health approach. PMID- 9854696 TI - Hiroshima and paper cranes: a technique to deal with death and grief. PMID- 9854697 TI - Exploratory and confirmatory factor analyses of the Brief Symptom Inventory among substance abusers. AB - The Brief Symptom Inventory (BSI) is a multidimensional measure of psychological and somatic distress that is used to obtain detailed symptom profiles. The BSI has been questioned regarding its ability to differentiate among its proposed nine dimensions, and the factor structure underlying the BSI has not been confirmed with substance abusers. Exploratory factor analyses were completed on substance abusers (ntotal = 453; nwomen = 121; nmen = 332); nine factors were not identified. Five models were then submitted to confirmatory factor analyses using an independent sample of substance abusers (ntotal = 456; nwomen = 127; nmen = 329). A one-factor model (i.e., global psychological distress) best represented the data. Implications for using the BSI in research and counseling are discussed. PMID- 9854698 TI - Prescription and non-prescription drug use: a longitudinal study. AB - This study examines changes in the lifetime prevalence of prescription and non prescription drug use in a national longitudinal sample of young adults. Cohort data used in this study are from the National Longitudinal Survey, Youth Cohort (NLSY) who responded to questions on use of prescription drugs for the years 1984 and 1992 (N = 8,771). Results reveal increases in the lifetime prevalence of prescription use of sedatives, transquilizers, and stimulants. Prescription use was higher among women while non-prescription use was higher among men. Non prescription drug use in 1984 was significantly related to prescription use in 1992. PMID- 9854699 TI - Symptoms and characteristics of individuals with different types of recovery from DSM-IV alcohol dependence. AB - Symptoms and criteria for DSM-IV alcohol dependence and demographic and drinking characteristics are compared for three groups. Group I (N = 1,044) consists of persons who formerly met the criteria for dependence but were abstinent in the past year, Group II (N = 2,325) consists of persons who were formerly dependent but did not meet the criteria for abuse or dependence in the past year despite drinking, and Group III (N = 22,204) consists of persons who never met the criteria for dependence. Members of Group II lay between members of Groups I and III in terms of the numbers of prior dependence symptoms and criteria, past level of intake, degree of familial alcoholism and history of alcohol treatment. Both groups of former alcoholics were equally likely to have experienced withdrawal, drinking more/longer than intended and tolerance. The criteria that most strongly distinguished the groups were continued use despite physical or psychological consequences, time spent drinking and activities given up, all of which were far more common in Group I than in Group II. Members of Group II had earlier onsets of heavy drinking and dependence than members of Group I, supporting the existence of a developmentally limited subtype of alcoholism that is subject to spontaneous remission in early adulthood without treatment. The paper compares three options for tightening the criteria for dependence, all of which remove more members from Group II (28% to 41% depending on option) than from Group I (12% to 19%). PMID- 9854701 TI - Age of onset of drug use and its association with DSM-IV drug abuse and dependence: results from the National Longitudinal Alcohol Epidemiologic Survey. AB - The purpose of this study was to examine the relationship between early onset drug use and the development of lifetime DSM-IV drug abuse and dependence using a representative sample of the U.S. population. Prevalences of lifetime drug abuse and dependence were estimated for each year of age of onset of drug use from ages 13 and younger to 21 and older for the overall sample of drug users by race and gender. Linear logistic analyses were conducted to assess the relationship between age of drug use onset and lifetime drug use disorders controlling for important covariates. The major finding of this study was that early onset drug use is a significant predictor of the subsequent development of drug abuse over the life course. Early onset drug use was also a significant predictor of the subsequent development of lifetime alcohol dependence among males, females, and nonblacks, but not among blacks. After adjusting for important model covariates, the likelihood of lifetime drug abuse and dependence among the total sample of lifetime drug users was reduced by 4% and 5% with each year drug use onset was delayed. Implications of these findings are discussed in terms of the importance of collecting national data on drug use, abuse and dependence and the need for further research and its integration with prevention efforts. PMID- 9854700 TI - Smoked cocaine self-administration in females and voucher incentives for abstinence. AB - There are three purposes for this study: (1) To extend the laboratory study of heavy smoked cocaine use to women, (2) to assess cocaine withdrawal symptoms and (3) to assess the utility of voucher incentives for achieving and maintaining cocaine and other drug abstinence in female cocaine abusers. METHODS: Ten non treatment seeking female cocaine smokers resided inpatient for 4-5 days and could smoke up to 6 doses of cocaine base (50 mg each) twice a day (at 1200 h and again at 1600 h) for 2 consecutive days. During the following 2-week outpatient phase, women were given US $40 in merchandise vouchers if urinalysis indicated lower drug levels from the previous day. RESULTS: Women self-administered 20.4 out of 24 possible doses. Compared to the 1200 session, heart rate and blood pressure, but not subjective effects, were still significantly increased prior to the 1600 session. Nine women completed the outpatient phase, attending 98% of their appointments. Using the One-Half Rule, 56% of urines indicated no new cocaine or other drug use. IMPLICATIONS: Although a US $40 voucher incentive for a "clean" urine was not sufficient to eliminate cocaine use, the possibility of earning the voucher was sufficient to maintain nearly perfect attendance. PMID- 9854703 TI - Relationships of substance use to attitudes toward gender roles, family and cohabitation. AB - This study utilizes data from four nationally representative subsamples of white high school seniors to analyze the relationships of alcohol consumption, marijuana use, and other illicit drug use to attitudes toward gender roles, family, and cohabitation. These analyses provide evidence concerning two hypotheses: the Gender Roles Hypothesis and Deviance Hypothesis. As predicted by the Gender Roles Hypothesis, among males, traditional gender role attitudes were associated with greater alcohol consumption. Contrary to the predictions of the Gender Roles Hypothesis, for females, nontraditional gender role attitudes were not associated with greater alcohol consumption or drug use in analyses which controlled for confounding factors such as family structure, socioeconomic status, and religiosity. As predicted by the Deviance Hypothesis, nontraditional attitudes toward cohabitation and family were associated with greater alcohol consumption and illicit drug use among both males and females. PMID- 9854702 TI - Relationship between readiness for and processes of change in a sample of alcohol dependent males. AB - Previous research (Isenhart, 1997) has shown a relationship between pre-treatment stages of change and outcome. The objective of this project was to evaluate whether certain processes of change measures predicted stages of change measures. A stepwise multiple regression model used Contemplation, Determination, and Action scores as dependent variables and process of change scores and measures of alcohol dependence severity and pre-treatment alcohol consumption as independent variables. The only process of change scale consistently correlated with the Contemplation, Determination, and Action scales was Self-reevaluation. Self reevaluation was negatively correlated with Contemplation and positively correlated with Determination and Action. These results suggest that as patients become more discouraged and frustrated with their alcohol use, they experience less ambivalence, are more likely to acknowledge that a problem exists, and are more willing to take action to address that problem. Implications and limitations of these findings are provided and inconsistencies between previous studies and these results are discussed. PMID- 9854704 TI - Alcohol and aggression: general population views about causation and responsibility. AB - The purpose of this paper is to explore the public's perceptions about alcohol as a causal agent in aggressive behavior, and to assess how these beliefs are associated with notions of responsibility and the excuse-function of alcohol. In a 1995 probability survey, 994 adults across Ontario (50.3% female; mean age = 41.5, SD = 5.9) were asked questions about: alcohol-aggression expectancies; alcohol as an excuse; responsibility; personal drinking behavior; alcohol aggression victimization; and demographics. Descriptive and regression analyses were conducted. Over three-quarters of respondents believed that alcohol is associated with aggression, with females, older respondents, those with less education, and those who do not drink heavily more likely to hold this view. A majority (92%) believed that an intoxicated person is responsible for any behavior, with very little subgroup variation. Analyses showed that the perception of alcohol as a causal agent was not associated with decreased personal responsibility attributions. In fact, the stronger the belief in the alcohol-aggression link, the more likely one was to hold the view that an intoxicated person is responsible for behavior. Beliefs that alcohol causes violence do not translate into the acceptance of intoxication as an excuse. Reasons as to why intoxication does not alleviate responsibility for the drunken actor--a result inconsistent with attribution theory--are discussed. The consistency of these results with the "New Temperance" movement in the United States is also discussed. PMID- 9854705 TI - Combined cyclodialysis and cataract surgery. AB - BACKGROUND AND OBJECTIVE: Glaucoma patients who require surgery for cataract may be treated with a combined surgical procedure. The purpose of this study was to determine the effectiveness of combining cyclodialysis with self-sealing, small incision cataract surgery for patients with cataracts and glaucoma. PATIENTS AND METHODS: This was a retrospective study. Fifty consecutive patients who had undergone the combined surgical procedure in 1992 or 1993 were identified. The patients were observed through the medical charts to determine when treatment failure (lack of improvement in intraocular pressure [IOP] control) occurred. The routine clinic examination schedule for the cases was generally at 1 to 2 days, 1 to 2 weeks, 1 to 2 months, 3 to 4 months, 5 to 6 months, 8 to 9 months, 1 year, and at yearly intervals thereafter. The rates of treatment failure and mean time to failure were determined using life-table methods. IOP control status at the final available follow-up examination was summarized for the group. RESULTS: Thirty-seven of the 50 patients (74%) had a successful result. At the final available follow-up, 31 of these patients (62% of the entire cohort) were without all medications while maintaining IOP control. The other 6 patients demonstrated improved control by either requiring less medication to maintain control or by improving from the baseline level while being treated with medication. The reduced pressures appeared to be stable and maintained throughout a median follow up time of 29 months. CONCLUSION: Combining cyclodialysis with small incision cataract surgery has been successful in a high percentage of cases in the author's experience. It has become the procedure of first choice for patients with cataracts and glaucoma. PMID- 9854706 TI - Surgical outcome of transconjunctival cryosurgical extraction of orbital cavernous hemangioma. AB - BACKGROUND AND OBJECTIVE: Transconjunctival cryosurgical extraction (TCE) is not an accepted method for the removal of most intraorbital tumors, and thus is not routinely performed. This study evaluates TCE for orbital cavernous hemangioma (OCH), which, due to its specific vascular structure, is suitable for this technique. PATIENTS AND METHODS: In this retrospective case series, the medical charts of 12 consecutive patients with OCH who underwent TCE were reviewed. The operative complications and the surgical outcome were analyzed. RESULTS: Eleven tumors were intraconal and 1 was extraconal. No tumor involved the orbital apex. In 10 patients (83%), the operation was uneventful. One patient lost vision due to intraoperative cilioretinal arterial occlusion. In another patient, the tumor could not be identified and was subsequently removed via lateral orbitotomy. CONCLUSION: TCE, a relatively minor operation, is as safe as more extensive surgical procedures for the removal of OCH not located in the posterior third of the orbital space. PMID- 9854707 TI - Surgical outcome on combined procedures of lens extraction, intraocular lens implantation, and vitrectomy during removal of the epiretinal membrane. AB - BACKGROUND AND OBJECTIVE: The authors evaluated the outcome of combined procedures of lens extraction, intraocular lens implantation, and vitrectomy during procedures for the removal of the epiretinal membrane, and attempt to clarify which factors affected the outcome of surgery. PATIENTS AND METHODS: Surgery for the removal of the epiretinal membrane was performed and a 2-year follow-up program was maintained for 32 eyes at the authors' clinic during the past 5 years. The combined procedure was undertaken in 15 eyes, whereas a simple vitrectomy was performed in 17 eyes. Results were analyzed by postoperative ophthalmoscopic and slit-lamp examinations for visual acuity and complications. RESULTS: Six months after surgery, visual acuity improved similarly in both groups. But, 2 years after surgery, 11 eyes (65%) that underwent a simple vitrectomy showed decreased visual acuity due to the advancement of cataracts. The postoperative visual acuity at 6 months correlated to preoperative visual acuity, and the advancement of cataracts tended to correlate inversely to the age of the patients. Major complications did not occur in any of the cases. CONCLUSION: Earlier surgical management is better for treatment of the epiretinal membrane, and a combined procedure has advantages in cost reduction and is less troublesome, especially for elderly patients. PMID- 9854708 TI - The use of dextran after keratoplasty for visualizing the vitreous cavity. AB - BACKGROUND AND OBJECTIVE: Corneal edema during penetrating keratoplasty (PK) precludes the visualization of the fundus required to perform vitreoretinal surgery. In this study, the clearing of the cornea and fundus visualization with dextran 40 following PK was evaluated. PATIENTS AND METHODS: Topical dextran 40 was used in 12 eyes that underwent uncomplicated PK (phase 1) and in 5 eyes that underwent combined PK with vitreoretinal surgery (phase 2). RESULTS: Clearing of the cornea and visualization of the fundus were observed in all eyes after dextran had been instilled for a mean of 4 and 5 minutes in eyes for phase 1 and 2, respectively. Vitreoretinal surgery could successfully be performed after PK in all eyes of phase 2. CONCLUSION: Topical use of dextran 40 following PK produces enough clearing of the cornea to enable visualization of the fundus for further vitreoretinal procedures after the graft has been sutured. PMID- 9854709 TI - Prevention of posterior capsule opacification with the CO2 laser. AB - BACKGROUND AND OBJECTIVE: Posterior capsule opacification (PCO) is a common complication after cataract extraction, despite the modern surgical techniques and lenses being used for this procedure. Its prevention challenged many investigators, because the current treatment of choice, capsulotomy with Nd:YAG laser, is associated with sight-threatening complications. In the present study, the authors investigated two approaches of preventing PCO using the CO2 laser. MATERIALS AND METHODS: A 15-W CO2 laser with a 17- or 18-gauge hollow probe was used on 20 sheep eyes and 14 rabbit eyes. Lens extraction was done by phacoemulsification. In the equatorial treatment study, the anterior chamber was filled with either air or a viscoelastic substance, and laser burns were applied to the equator of the lens capsule and to the peripheral anterior capsule to destroy the epithelial cells. In the capsulotomy study, a primary posterior capsulotomy was created by delivering 1 to 3 laser shots to the capsule behind an implanted intraocular lens (IOL). RESULTS: The CO2 laser was satisfactory in sheep eyes after filling the anterior chamber with air. In rabbit eyes, however, it was technically impractical to work with air. Using a viscoelastic material to maintain the anterior chamber, the hollow probe of the CO2 laser becomes plugged up and therefore is unable to affect the ocular tissue. However, by combining viscoelastic and air pumping, both the destruction of the lens epithelial cells and the creation of a central posterior opening behind a capsular-fixated IOL was repeatedly achieved. CONCLUSION: Using the CO2 laser for destruction of lens epithelial cells and the creation of controlled posterior capsulotomy is feasible and practical. A different design of the probe (closed gauge) is required to enable it to operate clinically in a fluid or viscoelastic environment. PMID- 9854710 TI - An extruded encircling band straddling the cornea and corneal groove formation. AB - The authors report a rare consequence of an anteriorly migrated encircling silicone band. The extruded encircling band, which "cheesewired" forward through the superior and medical rectus insertions straddled the cornea firmly and formed a corneal groove. Corneal groove is an extremely rare complication of scleral buckling surgery. PMID- 9854711 TI - Cutaneous malignant melanoma metastatic to the eyelid. AB - Cutaneous malignant melanoma metastatic to the choroid, orbit, or vitreous is rare. Metastatic lesions to the eyelid are rare as well. The authors report a case of malignant melanoma metastatic to the eyelid. This was the first sign of tumor recurrence in a patient who was thought to have been successfully treated for a melanoma of the back 3 years previously. The patient died 6 weeks after diagnosis of the eyelid lesion. In the presence of known malignancy, metastasis must be suspected in new eyelid lesions. PMID- 9854712 TI - Proliferative diabetic retinopathy with a proliferative membrane that spread anteriorly from the optic disc. AB - The authors report a case of proliferative diabetic retinopathy in which the proliferating membrane spread from the optic disc to the anterior of the eyeball, and did not appear to involve the posterior precortical vitreous pocket. PMID- 9854713 TI - Nd:YAG laser treatment for premacular hemorrhage. AB - The authors describe two cases of premacular hemorrhage secondary to retinal macroaneurysm, with significant loss of vision. A few low-energy bursts of ND:YAG laser were applied to perforate the anterior surface of the hemorrhage, subsequently draining the blood into the vitreous cavity. The blood gradually cleared from the vitreous, with restoration of good vision. Nd:YAG laser seems to be a safe and efficient method for treating premacular hemorrhage and should be considered as a therapeutic option. PMID- 9854714 TI - The iris cerclage suture for permanent mydriasis: a running suture technique. AB - Permanent mydriasis can be a disabling condition. The author describes an iris cerclage running suture technique for the treatment of permanent mydriasis caused by diffuse iris sphincter damage. A long, curved transchamber needle on 10-0 polypropylene is passed in and out of the anterior chamber through limbal paracentesis openings while weaving the needle through the iris near the pupillary margin to form the cerclage. The technique minimizes stretching and distortion of the iris and creates a pupil that is precisely sized and rather round. PMID- 9854715 TI - Rapid preparation of small-volume autologous fibrinogen concentrate and its same day use in bleb leaks after glaucoma filtration surgery. AB - The authors evaluated small-volume preparation of autologous fibrin glue (AFG) and same day use in postglaucoma filtration surgery patients with Seidel positive bleb leaks and determined fibrinogen concentrations in autologous fibrinogen concentrates (AFCs) from 10 volunteers. Thirty milliliters of blood was centrifuged (5 min, 2400 x g); plasma was frozen (5 min-ethanol and ice), thawed (1-6 C, 30-60 min), and centrifuged (10 min, 5 C, 2800 x g); and the precipitate was transferred to a 1.0-ml tuberculosis syringe. Thrombin (1000 U) was dissolved (0.8 sterile water, 0.2 ml aminocaproic acid) and warmed (37 C). Average preparation time was 90 minutes. Alternating drops of AFC and thrombin were applied to bleb leaks until AFC clotted. Seidel testing with fluorescein determined success. AFC was prepared from 10 volunteers and fibrinogen was measured. AFG was initially successful with two (Seidel negative) eyes; one eye remained negative. AFG was unsuccessful in one briskly Seidel-positive leak. Mean +/- SD fibrinogen concentration in AFCs from the 10 volunteers was 2314 +/- 643 mg/dl (range 1608-3431 mg/dl). AFG may successfully close bleb leaks in outpatient settings. Brisk aqueous flow may impair effectiveness of AFG. Fibrinogen concentrations were comparable with previous reports. PMID- 9854716 TI - Analysis of fetal sex in TCC sample DNA: a contribution to the validation of this approach. AB - We compared two methods of collection of transcervical cell samples, mucus aspiration and cytobrush, with respect to the efficiency in determining fetal sex and we correlated the results with the week of gestation (7-11 weeks) to evaluate if the age of gestation influenced the success of the analysis. DNA extracted from TCC samples recovered by mucus aspiration (n = 27) and cytobrush (n = 36) were analysed by nested PCR to predict fetal sex. The statistical indices of sensitivity, specificity, positive predictive value and negative predictive value were determined, and compared with those of other studies previously performed. No statistically significant difference was found between the two methods of TCC sampling concerning the success of fetal sex prediction which was high for both methods (78 per cent and 89 per cent) and no correlation with the week of gestation was found. Transcervical cell sampling represents an encouraging prospect for first-trimester prenatal diagnosis even when the least invasive techniques are used. PMID- 9854717 TI - First report of prenatal biochemical diagnosis of Lowe syndrome. AB - The oculocerebrorenal syndrome of Lowe (OCRL) is a rare X-linked disorder with a severe phenotype characterized by congenital cataracts, renal tubular dysfunction and neurological deficits. The gene has been characterized and mutations have been identified in patients. Owing to the allelic heterogeneity exhibited by this gene, prenatal diagnosis by molecular analysis is limited to families in which the mutation is already known or in which linkage is informative. A more generally applicable diagnostic test would be valuable for families at risk for Lowe syndrome. Since ocrl1 is now known to encode a phosphatidylinositol 4,5 bisphosphate 5-phosphatase (Ptdlns(4,5)P2 phosphatase), we assessed whether biochemical testing could be used for prenatal diagnosis. We report here the first case of prenatal diagnosis for Lowe syndrome by measuring phosphatidylinositol 4,5-bisphosphate 5-phosphatase activity in cultured amniocytes. PMID- 9854718 TI - Inhibin A and pro-alpha C inhibin in Down syndrome and normal pregnancies. AB - Using two enzyme-linked immunosorbent assays specific for inhibin A and pro-alpha C inhibin, levels of the two proteins were assessed in maternal serum from 43 Down syndrome and 300 chromosomally normal pregnancies at 15-17 weeks' gestation. Compared to the control pregnancies, both inhibin A and pro-alpha C inhibin were significantly elevated in the Down syndrome pregnancies with median levels, expressed as multiples of the normal median, of 1.53 MoM and 1.34 MoM, respectively (P < 0.001 and P = 0.046 compared to controls). Levels of inhibin A and pro-alpha C inhibin were weakly but significantly correlated in both the control and the Down syndrome sera (r = 0.25, P < 0.0001; r = 0.4, P = 0.008, respectively). These data suggest that the mechanism(s) underlying the elevated inhibin levels observed in Down syndrome may affect the regulation of both the inhibin alpha- and beta A-subunits. PMID- 9854719 TI - Effect of maternal age curve on the predicted detection rate in maternal serum screening for Down syndrome. AB - The effect of the choice of maternal age-specific prevalence curve on the model predicted Down syndrome detection rate was examined. All 19 published regression curves from 11 birth prevalence series in four meta-analyses were included. The detection rate for a five per cent false-positive rate was estimated for three combinations of markers. For free beta human chorionic gonadotropin and alpha fetoprotein the lowest predicted detection rate was 62.3 per cent and the highest 64.1 per cent, a range of 1.8 per cent. When unconjugated oestriol was added as a third marker it was 65.6-67.3 per cent, a 1.7 per cent range, and when inhibin A was the fourth marker the detection rate was 72.0-73.4 per cent, a 1.4 per cent range. The number of series included in the regression had the biggest effect: when the authors had used a subset thought to have the highest ascertainment the predicted detection rate generally increased. The type of regression equation used and restrictions on the age range over which the regression was performed were less important factors. The effect of the choice of curve on the predicted increase in detection achieved by incorporating additional markers was relatively small: 3.1-3.3 per cent for unconjugated oestriol and a further 6.1-6.5 per cent for inhibin A. This analysis shows that the model inaccuracy caused by the maternal age curve is not small but is unlikely to be large enough to influence Down syndrome screening policy decisions. PMID- 9854720 TI - Two-colour immunocytochemical staining of gamma (gamma) and epsilon (epsilon) type haemoglobin in fetal red cells. AB - We have developed a two-colour immunocytochemical staining method for the detection of fetal and embryonic haemoglobin in erythroid cells. The method was applied to study these haemoglobin types in fetal red cells. Specimens from fetal blood (10 weeks), cord blood and fetal liver (14 weeks) as well as chorionic villus samples (10-13 weeks) were stained for gamma and epsilon chains using CY3 and FITC labelled antibodies. Morphometric analysis was applied to determine cell size. Samples from organs involved in early embryonic development contained relatively large erythroblasts expressing the epsilon globin chain (megaloblasts); later in gestation the gamma chain was co-expressed by the same cells which ultimately became smaller and contained HbF (alpha 2 gamma 2) only. This phenomenon was confirmed in CVS samples in which all cell types were abundantly present. Since fetal erythroblasts are considered candidate cells for non-invasive prenatal diagnosis using FISH, we studied the phenotype of erythroblasts circulating in the maternal blood. The majority of erythroblasts in maternal blood appeared to be of the relatively small gamma globin-containing cell type. However, careful screening of the same maternal blood samples also revealed erythroblasts expressing epsilon or epsilon and gamma globins simultaneously, although at low frequency. Control specimens from non-pregnant women did not show nucleated red cells expressing either of the haemoglobin types. These observations may contribute to the better recognition of fetal cells in the maternal blood for prenatal diagnosis. PMID- 9854721 TI - Fetal liver position and perinatal outcome for congenital diaphragmatic hernia. AB - Despite advances in postnatal care, patients born with a congenital diaphragmatic hernia (CDH) suffer substantial morbidity and mortality. The present study was undertaken to determine the prognostic influence of prenatally-diagnosed liver herniation in the hemithorax in fetuses with CDH. The medical records of 48 patients evaluated for a prenatally-diagnosed left CDH were retrospectively reviewed. Patients were analysed according to the position of the liver by prenatal ultrasound; 32 fetuses had a major portion of the liver herniated into the left hemithorax ('liver up') and 16 had an intra-abdominal liver ('liver down'). Liver position was determined using colour-flow Doppler ultrasonography. There were two fetal deaths in the liver-up group and one in the liver-down group. The liver-up group more frequently required extracorporeal membrane oxygenation (ECMO) support (53 per cent) compared with the liver-down group (19 per cent). Postnatal survival was significantly less in the liver-up group (43 per cent) vs. the liver-down group (93 per cent). Fetuses with congenital diaphragmatic hernia and liver herniated into the hemithorax have a much worse prognosis than similarly afflicted fetuses without liver herniation. Prenatal ultrasonographic diagnosis of congenital diaphragmatic hernia allows for preparation for a critically ill newborn and aids in prenatal family counselling. PMID- 9854722 TI - Assessment of the clinical usefulness of the 'Queenan' chart versus the 'Liley' chart in predicting severity of rhesus iso-immunization. AB - In 1961 Liley developed a chart of changing amniotic fluid bilirubin levels (delta OD450) and gestation, with three zones delineating the severity of rhesus disease. This chart ranged from 27 to 40 weeks and was found to be clinically useful. Extrapolating the use of the Liley chart to earlier gestations however, was unsuccessful. Currently, cordocentesis is the only reliable means of assessing the fetal condition accurately prior to 27 weeks. In 1993, Queenan proposed a chart of delta OD450 from 14 to 40 weeks, with four zones to guide management. The aim of the current study is to assess the clinical usefulness of the 'Queenan' chart vs. the 'Liley' chart. There were 35 pregnancies affected by rhesus disease between 1990 and 1997 at the Mater Mothers' Hospital, Brisbane. The quantitative anti-D and delta OD450 levels obtained before intra-uterine transfusions were recorded. Each sample was labelled with the fetal condition at the time the sample was taken. Of the 72 delta OD450 samples, 36 (50 per cent) were performed before 27 weeks, and these included all four of the severely affected samples and 11 of the 13 moderately affected samples. The sensitivity of the Queenan chart in predicting the severely affected pregnancies was 100 per cent, with specificity of 79.4 per cent, positive predictive value of 22.2 per cent and negative predictive value of 100 per cent. For prediction of moderate/severely affected pregnancies, the sensitivity was 83.3 per cent, with specificity of 94.4 per cent, positive predictive value of 83.3 per cent and negative predictive value of 96.3 per cent. In conclusion, an delta OD450 chart which includes gestations less than 27 weeks is necessary with our changing caseload of rhesus iso-immunized patients, where severely affected pregnancies seemed to occur early and intervention even in moderately affected pregnancies seemed appropriate. We found that the Queenan chart is a suitable alternative to the Liley chart. PMID- 9854723 TI - MRI does not change fetal cardiotocographic parameters. AB - The aim of this study was to determine whether magnetic resonance imaging (MRI) has any effect on fetal cardiotocographic (CTG) parameters or movement incidence. Sixteen mothers were examined during the last trimester at 28-39 weeks (mean 33 weeks; SD 4) of gestation due to a suspected fetal anomaly found in antenatal ultrasonography (US). MR imaging was performed using Siemens Magnetom Vision 1.5 T equipment with a 25 mT/m peak gradient amplitude. T2-weighted images were produced with HASTE and TRUE-FISP sequences and T1-weighted images with a 2D FLASH sequence. A four-element phase-array coil was used as the receiver. Before and after MRI-examination, a computerized analysis of the fetal heart rate (FHR) was produced. Basal FHR, short-term variation (STV) and fetal movements were calculated. The mean basal FHR was 136 beats/min (SD 11.6) before, and 133 beats/min (SD 8.9) after (P = 0.158). Short-term variation was in the normal range for both CTG-tracings: mean 9.7 ms (SD 2.7) and 8.8 ms (SD 2.8) (P = 0.196). The median for fetal movements before MRI was 48/h, and after MRI 24.5/h (P = 0.98). MRI at high field strength with powerful gradients did not affect fetal heart activity or movement incidence. PMID- 9854724 TI - Management of congenital diaphragmatic hernia diagnosed prenatally: an update. PMID- 9854725 TI - Ultrasound diagnosis of fetal diaphragmatic hernia and complex congenital heart disease at 12 weeks' gestation--a case report. AB - Congenital diaphragmatic hernia is commonly associated with congenital heart disease. Their coexistence indicates a poor prognosis. Prenatal diagnosis of these conditions in early pregnancy allows the option of pregnancy termination. We present a case of left-sided fetal diaphragmatic hernia and complex congenital heart disease diagnosed by ultrasound examination at 12 weeks' gestation. PMID- 9854726 TI - Perlman syndrome--a cause of enlarged, hyperechogenic kidneys. AB - Perlman syndrome is a rare overgrowth syndrome, thought to be inherited in the autosomal recessive fashion. We describe the sonographic features in two cases, and review the literature. PMID- 9854727 TI - Prenatally detected paternal uniparental chromosome 13 isodisomy. AB - A 13q isodisomy in a balanced karyotype: 45,XY,-13,-13, + i(13)(q10) was found in cultured amniocytes studied because of advanced maternal age. The isochromosome was monocentric and a new mutation as both parents had normal chromosomes. Fetal blood was studied to exclude 13-trisomy mosaicism. All (100) lymphocytes studied had the same karyotype with i(13)(q10) as the amniocytes. To determine the origin of the isochromosome, six microsatellite markers from 13q were analysed: D13S175, D13S166, D13S162, AC224, COLAC1 and D13S122. The results indicated that the i(13)(q10) was of paternal origin and isodisomic. The father had a risk of 1/20 for being a carrier for an autosomal recessive, progressive brain disorder, variant late infantile neuronal ceroid lipofuscinosis (CLN5). The risk for the fetus for this disorder of chromosome 13 was excluded by haplotype analysis. A healthy child was born at week 40 of pregnancy, supporting the idea that there are no paternally imprinted genes on chromosome 13q. Analysis of extra embryonal tissue (four samples studied) revealed the same balanced karyotype with the i(13)(q10)pat chromosome. According to the cytogenetic and molecular studies, the origin of the isochromosome 13 could be a transverse centromere cleavage at the paternal meiosis II or at an early mitosis. PMID- 9854728 TI - FISH analysis of a complex chromosome rearrangement involving nine breakpoints on chromosomes 6, 12, 14 and 16. AB - We report the prenatal diagnosis of an apparently balanced de novo complex chromosome rearrangement (CCR) which involved nine breakpoints on four different chromosomes. Fluorescence in situ hybridization (FISH) and spectral karyotyping (SKY) were performed as an adjunct to G-banding for characterization of the abnormal chromosomes. The 22-week female fetus showed minor dysmorphic features including dolichocephaly, broad fingernails, tibial bowing, clubfoot, thoracolumbar scoliosis and hypoplastic toenails. Autopsy revealed gall-bladder hypoplasia and an atrial septal defect. Chromosome analysis of fetal tissue confirmed the presence of the complex rearrangement. PMID- 9854729 TI - Poly-FISH: a technique of repeated hybridizations that improves cytogenetic analysis of fetal cells in maternal blood. AB - Prenatal diagnosis of fetal chromosomal abnormalities using interphase fetal nucleated erythrocytes (FNRBCs) separated from maternal peripheral blood can be technically challenging due to the limited number of FNRBCs available for analysis, the limited number of probes that can be used simultaneously, and low FISH efficiency on the formaldehyde-fixed and immunohistochemically stained interphase FNRBCs. We developed a technique of sequential FISH analysis that involves removal of the previous hybridized probe under denaturing conditions, and rehybridization with different probes to improve FISH efficiency. This technique facilitates the analysis of multiple chromosome-specific probes on the same nuclei. Results from our experiments show that FISH can be performed at least nine times on the same interphase nucleus and at least three different probes can be used simultaneously. Thus, theoretically, at least 24 different chromosomes can be analysed on a single interphase fetal cell isolated from maternal blood. We have termed this technique 'Poly-FISH', and have successfully diagnosed trisomy 21, triploidy, and other chromosome abnormalities in FNRBCs using this technique. PMID- 9854730 TI - Calcification of the fetal heart--four case reports and a literature review. AB - Calcification of the heart and vessels in fetuses is a rare condition. It may be dystrophic or metastatic. An extremely rare form of vascular calcification has been termed 'idiopathic arterial calcification of infancy', which is inherited in an autosomal recessive pattern. We report four cases of myocardial calcifications of different origin diagnosed in utero. The correct diagnosis is very important in regard to genetic counselling. PMID- 9854731 TI - Further observations of true mosaic trisomy 17 ascertained in amniotic fluid cell cultures. AB - Three new cases of true mosaic trisomy 17 (MT17) were diagnosed in amniotic fluid cells. Postnatal chromosome analysis from lymphocytes did not confirm the trisomic cell line, and follow-up studies showed normal psycho-motor development of the children, in one case up to the age of 4 1/2 years. We suggest that there are similarities between MT17 and MT20, in which the majority of pregnancies result in deliveries of healthy babies. PMID- 9854732 TI - Fetal enterolithiasis and anhydramnios; due to in utero hepatorenal syndrome? AB - Enterolithiasis is a rare, prenatal ultrasonographic finding. Previously reported cases were invariably associated with major fetal malformations. We describe a case of fetal enterolithiasis and anhydramnios in an anatomically normal fetus who, at autopsy, showed end-stage fetal liver disease. Fetal hepatorenal syndrome is the most probable cause of this in utero sonographic combination. PMID- 9854733 TI - Transvaginal sonographic diagnosis of short-rib polydactyly dysplasia at 13 weeks' gestation. AB - Short-rib polydactyly dysplasia (SRP) is an autosomal recessive, lethal skeletal dysplasia. Sonographic assessment of subsequent pregnancies is, therefore, recommended. This case indicates that this diagnosis can be made in the latter part of the first trimester. A 30-year-old multigravid woman presented at 13 weeks' gestation for an ultrasound examination. She had had a termination of pregnancy for a fetus with pathologically confirmed short-rib polydactyly dysplasia, type I (Saldino-Noonan). On transvaginal sonography, a narrow chest, symmetrical micromelia, polydactyly and anasarca were present. An autopsy confirmed recurrent SRP. Short-rib polydactyly dysplasia may be diagnosed with transvaginal sonography in the first trimester. PMID- 9854734 TI - Confined placental mosaicism for trisomy 8 and intra-uterine growth retardation. AB - This report describes a case of apparent confined placental mosaicism for trisomy 8 in a pregnancy which produced a male infant with intra-uterine growth retardation. Postnatal cytogenetic and molecular studies were consistent with biparental disomy 8. Postnatally, the infant experienced a period of rapid catch up growth and exhibited no clinical features of trisomy 8 mosaicism. His development was age appropriate. PMID- 9854735 TI - Prenatal diagnosis of fetal malformations by ultrafast magnetic resonance imaging. AB - Prenatal ultrasonography is the primary imaging modality in pregnancy as it allows direct real-time fetal examination. Antenatal magnetic resonance imaging (MRI) has so far been of limited clinical value owing to poor image quality. This was due to the long acquisition times that were needed to achieve a high enough spatial resolution for assessment of the small fetal anatomic structures resulting in severe motion artefacts. This problem has now been overcome by recent technical improvements. We present a case of fetal malformation (MURCS association), where an ultrafast T2-weighted RARE sequence (single shot fast spin echo sequence) provided images of excellent quality, which led to the correct antenatal diagnosis. MRI using this sequence can now be used for prenatal diagnosis and is thus likely to become an important adjunct to ultrasonography, especially in cases in which ultrasound findings are unclear of sonographic images are impaired by maternal obesity or oligohydramnios. PMID- 9854736 TI - Prenatal diagnosis of cardiac osteoma: a case report. AB - Primary benign intracardiac tumours in the infant period are rare. We report a case of a cardiac osteoma detected at 17 weeks of gestation. Ultrasonographically, it appeared as a calcified mass with a sharp margin and was associated with hypoplastic right ventricle. The gross and histological findings are presented. PMID- 9854738 TI - Diagnosis and management of non-immune hydrops fetalis. PMID- 9854737 TI - Low second-trimester maternal serum human chorionic gonadotrophin in a trisomy 9 pregnancy. PMID- 9854740 TI - Current awareness in prenatal diagnosis. PMID- 9854739 TI - The right to fetal life. PMID- 9854741 TI - Stress and the developing limbic-hypothalamic-pituitary-adrenal axis. AB - The postnatal limbic-hypothalamic-pituitary-adrenal (LHPA) axis in the rodent is remarkably different from the adult, both in structure and function. The first 2 weeks postnatally are characterized by a 'silent period' during which the developing animal is hyporesponsive to stress (stress hyporesponsive period SHRP), followed by a new and unique phase of stress responsiveness when the animal fails to swiftly terminate glucocorticoid secretion. In this review, we summarize our work which focuses on the regulatory biology of the components of the LHPA system and the consequences of its disruption on the adaptive responses of the developing organism. We find that the animal during the first 2 weeks of life responds to an intermittent chronic challenge increasing anterior pituitary POMC post-translational events, while the adult increases genomic events. The result for both the mature and the developing animal is the same, an increase in corticosterone (CS) levels. In addition, we have found evidence of impaired rate sensitive feedback in the weanling animal, as well as changes in ACTH clearance. Similar to the young animal emerging from SHRP, maternally deprived pups during the first week of life exhibit a substantial and sustained ACTH and CS response to stress. In the deprived animal these changes are accompanied by decreases in mineralocorticoid receptor gene expression in the hippocampus, suggesting that changes in mineralocorticoid to glucocorticoid receptor ratios may be important in this phenomena. What has become evident from our studies is that mechanisms underlying normal LHPA development are dynamic, age dependent and distinct to the strategies used by the mature organism to cope with stress. PMID- 9854742 TI - Effect of diazepam and sex hormones on EEG of gonadectomized male and female rats. AB - Waking EEG was recorded from left and right parietals in 60 adult Wistar rats gonadectomized after puberty during 5 days, 1 as baseline, 1 with vehicle and 1 with 10 mg/kg i.m. of diazepam with hormonal treatment (either testosterone propionate, 5-alpha-dihydrotestosterone or vehicle in males and progesterone, estradiol benzoate or vehicle in females). Quantitative EEG analysis showed that: diazepam in gonadectomized males increased absolute power of alpha, beta 1 and beta 2, increased interparietal correlation of delta and decreased interparietal correlation of slow theta, fast theta and beta 1, whereas in gonadectomized females, diazepam additionally decreased slow and fast theta absolute power. EEG effects were not modified by testosterone propionate or 5-alpha dihydrotestosterone treatment in males. Diazepam plus estradiol rendered the female EEG response similar to the gonadectomized or testosterone-treated males response; diazepam plus progesterone induced the same response in females as in gonadectomized females and additionally induced interparietal asymmetry and decreased interparietal correlation of all EEG bands. Present results and information on the effects of diazepam on interparietal coupling of EEG and demonstrate that the EEG response to diazepam in adult rats is sexually dimorphic and depends on sex as well as on activational effects of gonadal steroids. PMID- 9854743 TI - Behavioral and neurobiological effects of estrogen replacement therapy and a history of triphasic oral contraceptive exposure. AB - The effects of contraceptive steroids and estrogen replacement therapy on behavior and neuroendocrine function were evaluated in adult female cynomolgus monkeys. During the 'premenopausal' phase of the experiment, the animals were assigned to either treatment with a triphasic oral contraceptive (OC) for 24 months or the untreated control group. The monkeys were then ovariectomized and half of each of the premenopausal groups were randomly assigned to either treatment with conjugated equine estrogens (ERT) or the untreated control group for 12 months (the 'postmenopausal' phase). All evaluations were completed during the postmenopausal phase of the experiment. Both types of exogenous steroid treatments appeared to increase cardiovascular and hypothalamic-pituitary-adrenal responses to stress in socially dominant but not socially subordinate females. A history of triphasic OC administration increased contact aggression received, and reduced the prolactin response to fenfluramine, suggesting reduced serotonergic activity, for at least a year following the cessation of triphasic OC treatment. PMID- 9854744 TI - Effects of the common cold on mood and performance. AB - Previous research has shown that both experimentally-induced and naturally occurring upper respiratory tract illnesses (URTIs) influence mood and mental functioning. None of the previous studies of naturally occurring colds has conducted appropriate virological assays to determine the nature of the infecting agent. This is an essential methodological step in studies of malaise associated with URTIs. The aim of this research was to investigate the effects of naturally occurring colds on mood and objective measures of performance. This was done by first conducting a cross-sectional comparison of 37 healthy people and 158 volunteers with colds and then a longitudinal study in which 100 volunteers developed colds and 87 remained healthy. Virological techniques were used to identify infecting agents and comparisons made across the different groups. The results showed that having a cold was associated with reduced alertness and slowed reaction times. These effects were observed both for colds where the infecting virus was identified and those where it was not. Similar effects were obtained for both rhinovirus and coronavirus colds. One may conclude that upper respiratory tract illnesses lead to a reduction in subjective alertness and impaired psychomotor functioning. This was true for both illnesses where the infecting agent was identified and for those clinical illnesses where no virus was detected. It is now important to identify the mechanisms linking infection and illness with the behavioural changes. Similarly, the impact of these effects on real-life activities such as driving needs examining. Finally, methods of treatment need to be developed which not only treat the local symptoms of the illnesses but remove the negative mood and the performance impairments. PMID- 9854745 TI - Baseline salivary cortisol levels and preconscious selective attention for threat. A pilot study. AB - This study was conducted to examine the relationship between baseline salivary cortisol (CORT) levels and selective attention for displays of angry faces. Selective attention was investigated using a pictorial emotional Stroop task, comparing colournaming-speed of angry and neutral faces. The task was assessed in supraliminal (unmasked) and subliminal (masked) conditions to 28 non-clinical subjects (14 male and 14 female). Repeated measures analysis of variance revealed a significant interaction between median split CORT levels (low vs. high) and masked face valence (angry vs. neutral). The latter effect was mainly due to significant facilitation in the high CORT subject-group; these subjects seemed to allocate their attention away from the masked angry face. A relation between baseline CORT levels and fast withdrawal behavior is suggested. PMID- 9854746 TI - Intragenic polymorphisms of the vitamin D receptor gene associated with intervertebral disc degeneration. AB - STUDY DESIGN: A study in genetic epidemiology of disc degeneration, based on lifetime exposure data, findings on magnetic resonance imaging, and genotyping of intragenic markers. OBJECTIVES: To pursue the potential correlation between common allelic variations in the vitamin D receptor locus and degeneration of the intervertebral disc. SUMMARY OF BACKGROUND DATA: Familial aggregation has been observed in intervertebral disc degeneration, but the relative significance of the genetic component and shared environmental influences is unknown. The identification of relevant candidate genes associated with disc degeneration would specify a genetic component and increase our understanding of the etiopathogenesis of disc degeneration. METHODS: From the population-based Finnish Twin cohort, 85 pairs of male monozygotic twins were selected based on exposure to suspected risk factors for disc degeneration. Interview data were gathered on relevant lifetime exposures, and thoracic and lumbar disc degeneration was determined through quantitative and qualitative assessments of signal intensity on magnetic resonance imaging, and qualitative assessments of disc bulging and disc height narrowing. Possible associations were examined between disc degeneration measures and two polymorphisms of the coding region of the vitamin D receptor locus. RESULTS: Two intragenic polymorphisms of the vitamin D receptor gene revealed an association with disc degeneration. Quantitatively assessed signal intensities of thoracic and lumbar (T6-S1) discs were 12.9% worse in men with the Taql tt genotype and 4.5% worse in men with the Tt genotype, compared with signal intensity in men with the TT genotype (age adjusted P = 0.003). A similar pattern was found between disc signal intensity and Fokl genotypes; men with the ff and Ff genotypes had mean signal intensities that were 9.3% and 4.3% lower, respectively, than those in men with FF genotypes (age-adjusted P = 0.006). The summary scores of qualitatively assessed signal intensity, bulging, and disc height were 4.0% and 6.9% worse in men with Ff and ff genotypes, respectively, when compared with those in men with the FF genotype (age-adjusted P = 0.029). CONCLUSION: Specific vitamin D receptor alleles were associated with intervertebral disc degeneration as measured by T2-weighted signal intensity, demonstrating for the first time, the existence of genetic susceptibility to this progressive, age-related degenerative process. PMID- 9854747 TI - Lumbar spine fusion by local gene therapy with a cDNA encoding a novel osteoinductive protein (LMP-1). AB - STUDY DESIGN: A posterior arthrodesis animal model using local expression of a newly discovered osteoinductive protein delivered in bone marrow cells. OBJECTIVE: To introduce the concept of local gene therapy and determine its feasibility for achieving lumbar spine fusion using a gene for a novel osteoinductive protein: LIM Mineralization Protein-1 (LMP-1). SUMMARY OF BACKGROUND DATA: Extensive work is currently underway to improve the healing success and morbidity associated with the gold standard bone-grafting material of autogenous iliac crest. As a result, alternative osteoinductive proteins and new delivery methods warrant investigation. The authors' laboratory recently identified a novel gene that had osteoinductive capacity in vitro and is therefore a candidate for a new in vivo osteoinductive agent. METHODS: Single level posterior lumbar and thoracic arthrodesis was attempted in 14 athymic rats. The graft material, which consisted of a devitalized bone matrix (no osteoinductive activity) soaked with 0.75 to 1.5 million bone marrow cells, was inserted with the dorsal spine exposed. In each rat, one site received marrow cells transfected with the cDNA encoding a novel osteoinductive protein. At the other site for a control, the marrow cells were transfected with the reverse copy of the cDNA that did not express any protein. Transfection of marrow cells for 2 hours was accomplished using the mammalian expression vector pCMV2. Rats were killed after 4 weeks, and the spines were evaluated by manual palpation, radiographs, and nondecalcified histology. RESULTS: In the pivotal experiment, successful spine fusion was obtained in 9/9 (100%) of the sites receiving marrow cells transfected with the active LMP-1 cDNA and in 0/9 (0%) of the sites receiving marrow cells transfected with the reverse (inactive) LMP-1 cDNA. Radiographs and histology confirmed the manual palpation results, demonstrating controlled new bone formation in the carrier and marrow transfected with the active LMP-1 cDNA and essentially no bone induction in the sites treated with marrow cells that did not express the protein. CONCLUSIONS: These data confirm that local delivery of the novel LMP-1 cDNA using bone marrow cells is feasible in vivo. Furthermore, these results demonstrate that posterior thoracic or lumbar spine fusion can be achieved in rats by local delivery of the LMP-1 cDNA. PMID- 9854748 TI - Compression-induced degeneration of the intervertebral disc: an in vivo mouse model and finite-element study. AB - STUDY DESIGN: An in vivo study of the biologic and biomechanical consequences of static compressive loading on the mouse tail intervertebral disc. OBJECTIVES: To determine whether static compression in vivo alters the biologic activity of the disc and leads to diminished biomechanical performance. SUMMARY OF BACKGROUND DATA: Static compressive stress that exceeds the disc's swelling pressure is known to change hydration and the intradiscal stress distribution. Alterations in hydration and stress have been associated with changes in disc cell activity in vitro and in other collagenous tissues in vivo. METHODS: Mouse tail discs were loaded in vivo with an external compression device. After 1 week at one of three different stress levels, the discs were analyzed for their biomechanical performance, morphology, cell activity, and cell viability. A second group of mice were allowed to recuperate for 1 month after the 1-week loading protocol to assess the disc's ability to recover. As an aid to interpreting the histologic and biologic data, finite-element analysis was used to predict region-specific changes in tissue stress caused by the static loading regimen. RESULTS: With increasing compressive stress, the inner and middle anulus became progressively more disorganized, and the percentage of cells undergoing apoptosis increased. The expression of Type II collagen was suppressed at all levels of stress, whereas the expression of aggrecan decreased at the highest stress levels in apparent proportion to the decreased nuclear cellularity. Compression for 1 week did not affect the disc bending stiffness or strength but did increase the neutral zone by 33%. As suggested by the finite-element model, during sustained compression, tension is maintained in the outer anulus and lost in the inner and middle regions where the hydrostatic stress was predicted to increased nearly 10 fold. Discs loaded at the lowest stress recovered anular architecture but not cellularity after 1 month of recuperation. Discs loaded at the highest stress did not recover anular architecture, displaying islands of cartilage cells in the middle anulus at sites previously populated by fibroblasts. CONCLUSIONS: The results of the current project demonstrate that static compressive loading initiates a number of harmful responses in a dose-dependent way: disorganization of the anulus fibrosus; an increase in apoptosis and associated loss of cellularity; and down regulation of collagen II and aggrecan gene expression. The finite element model used in this study predicts loss of collagen fiber tension and increased matrix hydrostatic stress in those anular regions observed to undergo programmed cell death after 1 week of loading and ultimately become populated by chondrocytes after one month of recuperation. This correspondence conforms with the suggestions of others that the cellular phenotype in collagenous tissues is sensitive to the dominant type of tissue stress. Although the specific mechanisms by which alterations in tissue stress lead to apoptosis and variation in cell phenotype remain to be identified, our results suggest that maintenance of appropriate stress within the disc may be an important basis for strategies to mitigate disc degeneration and initiate disc repair. PMID- 9854749 TI - Psychosocial job factors, physical workload, and incidence of work-related spinal injury: a 5-year prospective study of urban transit operators. AB - STUDY DESIGN: Five-year prospective cohort study of 1449 transit operators. OBJECTIVES: To investigate psychosocial job factors as predictors of work-related spinal injuries, controlling for current and past physical workload. SUMMARY OF BACKGROUND DATA: The association between psychosocial job factors and spinal disorders may be confounded by physical workload. A 1991 prospective study of Boeing workers found psychosocial but not physical factors to be associated with spinal injuries. However, data on physical workload were limited. Recent cross sectional studies of transit drivers showed both physical and psychosocial factors to be independently associated with back and neck pain. This study was designed to test these findings prospectively. METHODS: Spinal injuries were ascertained from workers' compensation records, employment history from company records, and psychosocial factors from questionnaires. Logistic regression models adjusted for age, gender, height, weight, vehicle type, and current and past physical workload. RESULTS: During follow-up, 320 drivers reported a first spinal injury. Spinal injury was predicted by psychological job demands (odds ratio [OR], 1.50; 95% confidence interval [CI], 1.33-1.95); job dissatisfaction (OR, 1.56; 95% CI, 1.09-2.23); and the frequency of job problems (OR, 1.52; 95% CI, 1.02-2.26). Marginally significant associations were found for low supervisor support (OR, 1.30; 95% CI, 0.99-1.72) and female gender (OR, 1.49; 95% CI, 0.95 2.32). Compared with full-time work, part-time work was associated with a 2.7 fold reduced risk for spinal injury (OR, 0.37; 95% CI, 0.15-0.93). Cable car crews performing the heaviest physical labor had a threefold increased risk of spinal injury compared with bus drivers (OR, 3.04; 95% CI, 1.85-5.00). CONCLUSIONS: Physical workload and psychosocial job factors both independently predict spinal injury in transit vehicle operators. PMID- 9854750 TI - Acute effects of nucleus pulposus on blood flow and endoneurial fluid pressure in rat dorsal root ganglia. AB - STUDY DESIGN: An experimental study to elucidate the initial factors in the pathogenesis of lumbar pain caused by disc herniation. OBJECTIVE: To evaluate the effects of autologous nucleus pulposus on blood flow and endoneurial fluid pressure in dorsal root ganglia. SUMMARY OF BACKGROUND DATA: Human sciatica is known to be associated with compression of lumbar nerve roots and dorsal root ganglia by herniated intervertebral discs. Recently, it has been shown that application of nucleus pulposus to nerve roots induces injury and pain-related behavior in experimental animals. In this study, the authors hypothesized that nucleus pulposus applied to a nerve root would cause increased intraneural edema and reduced blood flow in the corresponding dorsal root ganglia. Studies in peripheral nerves have shown that these initial pathophysiologic disturbances initiate complex events that exacerbate nerve injury and cause pain. METHODS: A total of 29 adult female Sprague-Dawley rats weighing 200 to 250 g had their left L5 nerve roots and associated dorsal root ganglia exposed. Autologous nucleus pulposus was harvested from the tail and applied to the L5 nerve root just proximally to the dorsal root ganglia (nucleus pulposus group). For control, the same volume of muscle was harvested from the surgical area in the back and applied similarly to the neural tissue (control group). Blood flow was continuously monitored using a laser Doppler flow probe for 3 hours (n = 10) or 4 hours (n = 8) in animals with indwelling cannulas for measurement of systemic arterial pressure. Endoneurial fluid pressures were recorded with a servonull micropipette system using glass micropipettes with tip diameters of 4 microns. Endoneurial fluid pressure in the dorsal root ganglia was measured before and 3 hours after application of nucleus pulposus (n = 7) or muscle (n = 4). After measurement of blood flow and endoneurial fluid pressure, the nerve root and dorsal root ganglia were processed for histology and evaluated by light microscope. RESULTS: Blood flow in the nucleus pulposus group was reduced by 10% to 20% from the initial value after 3 to 4 hours. This reduction was statistically significant compared with that of the control group (P < 0.01). Endoneurial fluid pressure was initially 2.6 +/- 1.2 cm H2O in the nucleus pulposus group, and 2.1 +/- 0.6 cm H2O in the control group. Three hours after application, endoneurial fluid pressure was 7.5 +/- 4.6 in the nucleus pulposus group (P > 0.05), and 2.0 +/- 0.8 in the control group (P > 0.05). Edema was the principal pathologic finding seen consistently in the nerve roots and in many of the associated dorsal root ganglia from animals treated with nucleus pulposus. CONCLUSION: Application of nucleus pulposus to nerve root increased endoneurial fluid pressure and decreased blood flow in the dorsal root ganglia. This study's acute observations in the dorsal root ganglia may thus help to explain why disc herniations without compression of neural tissue are sometimes painful because similar pathologic findings are observed after only nucleus pulposus application to the nerve root. The authors further suggest that exposure of nerve roots to nucleus pulposus may establish a "compartment syndrome" in the dorsal root ganglia. PMID- 9854751 TI - The effect of compressive force applied to the intervertebral disc in vivo. A study of proteoglycans and collagen. AB - STUDY DESIGN: Coil springs were stretched and attached to produce a compressive force across the lumbar intervertebral discs of dogs for up to 27 weeks. OBJECTIVE: To test the hypothesis that a high compressive force applied over a period of time affects the production of proteoglycans and collagen by the intervertebral disc cells. SUMMARY OF BACKGROUND DATA: It is a commonly held belief that high forces applied to the intervertebral disc, and to joints in general, play a role in causing degeneration. METHODS: Pairs of stainless steel coil springs were stretched and attached to produce a compressive force across the lumbar intervertebral discs (L1-L2 and L3-L4) of 16 dogs. Dogs were killed between 13 and 27 weeks after the springs were attached. The discs (L1-L2 and L3 L4) were excised and assessed using immunohistochemical analyses and enzyme linked immunosorbent assay; T13-L1 and L4-L5 were used as controls. RESULTS: The main result relates to a group effect in the six dogs, assessed using enzyme linked immunosorbent assay, that were generally at the highest values of force for the greatest number of weeks. For the nucleus, but not the anulus, Spearman rank correlations revealed a strong correlation between increases in force and force-weeks (force multiplied by number of weeks) and increases in collagen type I accompanied by decreases in proteoglycans, chondroitin sulfate, and collagen type II for both experimental discs (L1-L2 and L3-L4), as compared with corresponding values in the controls (T13-L1 and L4-L5). In other words, as either the force or the force-weeks increased, the effect on the nucleus became greater. CONCLUSION: A high compressive force applied to the disc over a period of time initiates changes in proteoglycans and collagen. PMID- 9854752 TI - Tumor necrosis factor alpha and nucleus-pulposus-induced nerve root injury. AB - STUDY DESIGN: The effects of nucleus pulposus and various treatments to block tumor necrosis factor alpha activity were evaluated in an experimental set-up using immunohistochemistry and nerve conduction velocity recordings. OBJECTIVES: To assess the presence of tumor necrosis factor alpha in pig nucleus pulposus cells, and to see if block of tumor necrosis factor alpha also blocks the nucleus pulposus-induced reduction of nerve root conduction velocity. SUMMARY AND BACKGROUND DATA: A meta-analysis of observed effects induced by nucleus pulposus revealed that these effects might relate to one specific cytokine-tumor necrosis factor alpha. METHODS: Series-1: Cultured nucleus pulposus cells were stained immunohistologically with a monoclonal antibody for tumor necrosis factor alpha. Series-2: Nucleus pulposus was harvested from lumbar discs and applied to the sacrococcygeal cauda equina in 13 pigs autologously. Four pigs received 100 mg of doxycycline intravenously; five pigs had a blocking monoclonal antibody to tumor necrosis factor alpha applied locally in the nucleus pulposus, and four pigs remained nontreated, forming a control group. Three days after the application, the nerve root conduction velocity was determined over the application zone by local electrical stimulation. RESULTS: Series-1: Tumor necrosis factor alpha was found to be present in the nucleus pulposus cells. Series-2: The selective antibody to tumor necrosis factor alpha limited the reduction of nerve conduction velocity, although in comparison with the control group this was not statistically significant. However, treatment with doxycycline significantly blocked the nucleus-pulposus-induced reduction of conduction velocity. CONCLUSION: For the first time, a specific substance, tumor necrosis factor alpha, has been linked to the nucleus-pulposus-induced effects of nerve roots after local application. Although the effects of this substance may be synergistic with those of other similar substances, the data of the current study may be of significant importance for the continued understanding of nucleus pulposus' biologic activity, and of possible potential use for future strategies in managing sciatica. PMID- 9854753 TI - Changes in collagen cross-linking in degenerative disc disease and scoliosis. AB - STUDY DESIGN: Biochemical study of the changes in the collagen cross-link profile of human intervertebral discs collected at surgery from patients with either low back pain associated with disc degeneration or scoliosis. OBJECTIVE: To determine whether changes occur in the collagen cross-link profile in the disc of patients with either low back pain associated with disc degeneration or scoliosis, which may well influence matrix integrity. Such changes in the cross-link profile of a tissue indicates increased matrix turnover and tissue remodeling and may have implications for the progression of these disorders. SUMMARY OF BACKGROUND DATA: The diseases of the intervertebral disc, degenerative disc disease and scoliosis, are both characterized by changes in the extracellular matrix components that will affect the mechanical function of the tissue. The stability of the collagenous components and hence the mechanical integrity of connective tissues such as the disc is dependent on the degree and type of cross-links between the collagen molecules. This article reports results on the distribution of the different cross-links in the disc and the changes that occur with age, degenerative disc disease, and scoliosis. METHODS: Thirty-three discs were obtained from patients with degenerative disc disease and 29 discs from patients with scoliosis. Samples were acid hydrolyzed and the collagen cross-links analyzed by either fractionation on an amino acid analyzer configured for cross link analysis using ninhydrin postcolumn detection or fractionation by high pressure liquid chromatography with fluorescence detection. RESULTS: The reducible cross-links and the mature cross-link all increased from the outer anulus fibrosus through into the nucleus pulposus. The highest levels of the mature cross-link were found in the cartilage end-plate. The nonenzymic derived cross-link, pentosidine, in contrast, showed little difference across the disc, but did show the expected age-related increase. In degenerative disc disease, no change in the levels of the reducible or mature cross-links was found, but a decrease was observed in the levels of the age-related cross-link pentosidine in the more severe disease samples. In scoliosis, significantly higher levels of the reducible cross-links were found on the convex than on the concave side of the scoliotic disc. CONCLUSIONS: These changes in the cross-link profile of the intervertebral disc in degenerative disc disease and scoliosis are indicative of increased matrix turnover and tissue remodeling and likely to have implications for the progression of these disorders. PMID- 9854754 TI - The ligamento-muscular stabilizing system of the spine. AB - STUDY DESIGN: Electrical and mechanical stimulation of the lumbar supraspinous ligament of three patients with L4-L5 spinal deficits and of the feline model, respectively, was applied while recording electromyography on the multifidus muscles. OBJECTIVES: To determine if mechanoreceptors in the human spine can reflexively recruit muscle force to stabilize the lumbar spine, and to demonstrate, in the feline model, that such ligamento-muscular synergy is elicited by mechanical deformation of the lumbar supraspinous ligament (and possibly of other spinal ligaments), the facet joint capsule, and the disc. SUMMARY OF BACKGROUND DATA: The literature repeatedly confirms that ligaments have only a minor mechanical role in maintaining spine stability, and that muscular co-contraction of anterior and posterior muscles is the major stabilizing mechanism of the spine. The literature also points out that various sensory receptors are present in spinal ligaments, and that the ligaments are innervated by spinal and autonomic nerves. Data that describe how ligaments and muscles interact to provide stability to the spine were not found. METHODS: The supraspinous ligament at L2-L3 and L3-L4 was electrically stimulated in three patients undergoing surgery to correct deficits at L4-L5. Electromyography was performed from the multifidus muscles at L2-L3 and L3-L4, bilaterally. In 12 cats, the supraspinous ligaments from L1-L2 to L6-L7 were mechanically deformed, sequentially, while electromyography was performed from the multifidus muscles of the six levels. Loading of the ligament was applied before and after each of the two vertebrae were externally fixed to prevent motion. RESULTS: Electromyograms were recorded from the multifidus muscles, bilaterally, in the two of the three patients, demonstrating a direct relationship to receptors in the supraspinous ligament. Electromyograms were recorded from the feline multifidus muscle with mechanical loading of the supraspinal ligament at each of the L1-L2 to L6-L7 motion segments. In the free-spine condition the largest electromyographic discharge was present in the level of ligament deformation, and lower electromyographic discharge was recorded in two rostral and caudal segments. After immobilizing any two vertebrae, loading of the ligament resulted in electromyographic discharge in the muscles of the same level and at least one level above and/or below. CONCLUSIONS: Deformation or stress in the supraspinous ligament, and possibly in other spinal ligaments, recruits multifidus muscle force to stiffen one to three lumbar motion segments and prevent instability. Strong muscular activity is seen when loads that can cause permanent damage to the ligament are applied, indicating that spastic muscle activity and possibly pain can be caused by ligament overloading. PMID- 9854755 TI - Estimation of trunk muscle forces and spinal loads during fatiguing repetitive trunk exertions. AB - STUDY DESIGN: The effects of human trunk extensor muscle fatigue on the estimated trunk muscle forces and spinal loading were investigated during the performance of repetitive dynamic trunk extension. OBJECTIVE: To evaluate if alterations in the trunk muscle recruitment patterns resulted in a greater estimated active loading of the spine and, in turn, an increased risk of injury. SUMMARY OF BACKGROUND DATA: Epidemiologic studies highlight the increased risk of low back injury during repetitive lifting, implicating fatigue of muscles and/or passive tissues as causes of such injury. Increased trunk muscle activity or altered recruitment patterns resulting from fatigue in the primary trunk extensor muscles may indicate an increase in the active loading of the spine, which could contribute to an increased risk of injury. METHODS: Sixteen healthy study participants performed repetitive isokinetic trunk extension endurance tests at two load levels and two repetition rates, while their net muscular torque output and trunk muscular activity were measured. During each exertion, trunk torque, position, and velocity were controlled, so that any change in muscle activity could be attributed to fatigue. An electromyography-assisted model, adapted to accommodate the decline in maximum muscular tension generation resulting from fatigue, was used to estimate the 10 trunk muscle forces and spinal loading. Linear regression was used to quantify the rate of change in muscle force and spinal loading resulting from fatigue, while analysis of variance was used to determine if the rate of change was dependent on the task conditions (load and repetition rate). RESULTS: Significant elevations were estimated for the latissimus dorsi and external oblique muscle forces in more than 70% of the endurance tests, whereas significant reductions in the erector spinae muscle force were predicted in 75% of the trials. The magnitude of the range of change of the erector spinae and latissimus dorsi muscle forces was dependent on the load level and repetition rate. The reduction in erector spinae forces offset the augmented force in the other muscles, because the net changes in compression and lateral shear forces on the spine were not significant, and the anteroposterior shear was reduced. CONCLUSION: The results of the study do not suggest that an increase in the muscular loading of the spine occurs as a result of changing trunk muscular recruitment patterns. Therefore, future studies should focus on injury mechanisms that may occur as a result of a change in the viscoelastic passive tissue responses, muscular insufficiency, or a decline in neuromuscular control and coordination. PMID- 9854756 TI - Injuries in the adolescent porcine spine exposed to mechanical compression. AB - STUDY DESIGN: An experimental porcine study in which functional lumbar spinal units were tested in compression to failure. Biomechanical, radiographic, magnetic resonance imaging, and histological characteristics are described. OBJECTIVES: To explain the different patterns of injury seen in adults and adolescents resulting from traumatic injury to the vertebrae and to explain the mechanism behind traumatic displacement of the ring apophysis seen in athletes. SUMMARY OF BACKGROUND DATA: Recent investigations of the spine in adolescent who have sustained trauma have shown injuries to the growth zone, whereas studies of adults have shown injuries to the vertebral body. A higher frequency of abnormalities in the discs, the vertebral bodies, the endplates, and the ring apophyses has been demonstrated in athletes with high loads on the spine. There is controversy over the etiology of these changes. METHODS: Twelve functional lumbar spinal units (vertebra-disc-vertebra) obtained from six young male pigs were tested in compression to failure. All units were examined with plain radiography and magnetic resonance imaging before and after compression. After the compression, histologic samples were taken from the injury site. RESULTS: Identical traumatic changes were seen in all functional lumbar spinal units, i.e., fracture in the endplate through the growth zone posteriorly and displacement of the anulus fibrosus with a bony fragment at the point of insertion of the vertebra. The nucleus pulposus was ruptured and displaced through the fracture line in all cases. The injuries were not seen on radiographs but were detected on magnetic resonance images, as confirmed on microscopic and histologic examination. CONCLUSION: This study shows that the weakest part of the lumbar spine of the juvenile pig, when compressed, is the growth zone and the junction between the point of insertion of the anulus fibrosus and the vertebra. This location of weakness may explain the high frequency of disc degeneration and "persisting apophysis" seen in the spine of athletes. PMID- 9854757 TI - Intramuscular pressure in the erector spinae and intra-abdominal pressure related to posture and load. AB - STUDY DESIGN: Intramuscular pressures in both Erectors Spinae and intra-abdominal pressures were measured during different holding tasks. OBJECTIVES: To investigate the potential for using intramuscular pressure measurements in both Erectors Spinae to better quantitate the role of muscles during different lifting tasks in vivo. SUMMARY OF BACKGROUND DATA: Intramuscular pressure and intra abdominal pressure were measured previously under isometric and dynamic conditions. However, no previous study systematically has addressed the relation between intramuscular and intra-abdominal pressures and different loads, tasks, and postures. METHODS: Intramuscular and intra-abdominal were measured simultaneously with microtip pressure transducers in 10 healthy volunteers performing 24 different static holding tasks. Tasks included different weights (10 kg and 20 kg), postures (squat or back lift), and positions of the weight. RESULTS: Intramuscular pressures were dependent on posture. Kyphotic back posture produced intramuscular pressures of 120-130 mm Hg, compared with the 10-25 mm Hg produced when volunteers were in the erect position (P < 0.001). Holding a 10-kg weight at the thighs close to the body produced significantly (P < 0.001) lower intramuscular pressures (25-32 mm Hg) than that produced by holding it 25 cm in front of the body (47-56 mm Hg). In all tasks, intramuscular pressures were significantly higher with the 20-kg weight than with the 10-kg weight (P < 0.001). Highest values (> 300 mm Hg) were measured when the 20-kg weight was held in the kyphotic posture above the floor and 25 cm away from the body. CONCLUSIONS: Intramuscular pressure measurements in the erector spinae seem to be a valuable tool for quantitating the role of back muscles during different lifting tasks. PMID- 9854758 TI - Opioid therapy for chronic noncancer back pain. A randomized prospective study. AB - STUDY DESIGN: A randomized, open, long-term, repeated-dose comparison of an anti inflammatory drug and two opioid regimens in 36 patients with back pain. OBJECTIVES: To examine the long-term safety and efficacy of chronic opioid therapy in a randomized trial of patients with back pain. METHODS: All participants underwent a 4-week washout period of no opioid medication before being randomly assigned to one of three treatment regimens for 16 weeks: 1) naproxen only, 2) set-dose oxycodone, or 3) titrated-dose oxycodone and sustained release morphine sulfate. All patients then were assigned to a titrated dose of opioids for 16 weeks and then gradually tapered off their medication for 12 weeks. Finally, all participants were monitored for a 1-month posttreatment washout period. Each patient was called once a week for a report on pain, activity, mood, medication, hours awake, and adverse effects and was monitored carefully for signs of abuse and noncompliance. RESULTS: Weekly reports during the experimental phase showed the titrated-dose group to have less pain (P < 0.001) and less emotional distress (P < 0.001) than the other two groups. Both opioid groups were significantly different from the naproxen-only group. During the titration phase, patients also reported significantly less pain and improved mood. Few differences were found in activity or hours asleep, or between average pretreatment and posttreatment phone-interview and questionnaire variables. No adverse events occurred, and only one participant showed signs of abuse behavior. CONCLUSIONS: The results suggest that opioid therapy has a positive effect on pain and mood but little effect on activity and sleep. Opioid therapy for chronic back pain was used without significant risk of abuse. However, tapered-off opioid treatment is palliative and without long-term benefit. PMID- 9854759 TI - The reduction of chronic nonspecific low back pain through the control of early morning lumbar flexion. A randomized controlled trial. AB - STUDY DESIGN: Eighteen-month, randomized controlled trial with partial crossover. OBJECTIVES: To test the hypothesis that the control of lumbar flexion in the early morning will significantly reduce chronic, nonspecific low back pain. SUMMARY OF BACKGROUND DATA: Previous studies have indicated an increased risk of low back pain with bending forward in the early morning, primarily because of increased fluid content in the intervertebral discs at that time. METHODS: After 6 months of recording baseline data, 85 subjects with persistent or recurring low back pain were randomly assigned to treatment and control groups. The treatment group received instruction in the control of early morning lumbar flexion. The control group received a sham treatment of six exercises shown to be ineffective in reducing low back pain. Six months later, the control group received the experimental treatment, Diaries were used to record daily levels of pain intensity, disability, impairment, and medication usage. RESULTS: Significant reductions in pain intensity (P < 0.01) were recorded for the treatment group, but not for the control group (point estimate, 33%; 95% confidence interval, 11 55%). After receiving the experimental treatment, the control group responded with similar reductions (P < 0.05). Significant reductions also were observed in total days in pain, disability, impairment, and medication usage. CONCLUSIONS: Controlling lumbar flexion in the early morning is a form of self-care with potential for reducing pain and costs associated with chronic, nonspecific low back pain. PMID- 9854760 TI - A randomized trial of a lay person-led self-management group intervention for back pain patients in primary care. AB - STUDY DESIGN: Randomized, controlled trial. OBJECTIVE: To evaluate a four-session self-management group intervention for patients with pain in primary care, led by trained lay persons with back pain. The intervention was designed to reduce patient worries, encourage self-care, and reduce activity limitations. BACKGROUND DATA: Randomized trials of educational interventions suggest that activating interventions may improve back pain outcomes. Expert opinion increasingly regards effective self-management of back pain as important in achieving good outcomes. In this study, an educational intervention designed to activate patients and support effective self-management was evaluated. METHODS: Six to 8 weeks after a primary care visit for back pain, patients were invited to participate in an educational program to improve back pain self-management. Those showing interest by returning a brief questionnaire became eligible for the study. Participants (n = 255) randomly were assigned to either a self-management group intervention or to a usual care control group. The effect of the intervention, relative to usual care, was assessed 3, 6, and 12 months after randomization, controlling for baseline values. The intervention consisted of a four-session group applying problem-solving techniques to back pain self-management, supplemented by educational materials (book and videos) supporting active management of back pain. The groups were led by lay persons trained to implement a fully structured group protocol. The control group received usual care, supplemented by a book on back pain care. RESULTS: Participants randomly assigned to the self-management groups reported significantly less worry about back pain and expressed more confidence in self-care. Roland Disability Questionnaire Scores were significantly lower among participants in the self-management groups relative to the usual care controls at 6 months (P = 0.007), and this difference was sustained at 12 months at borderline significance levels (P = 0.09). Among self management group participants, 48% showed a 50% or greater reduction in Roland Disability Questionnaire Score at 6 months, compared with 33% among the usual care controls. CONCLUSIONS: Self-management groups led by trained lay persons following a structured protocol were more effective than usual care in reducing worries, producing positive attitudes toward self-care, and reducing activity limitations among patients with back pain in primary care. PMID- 9854761 TI - Efficiency and costs of medical exercise therapy, conventional physiotherapy, and self-exercise in patients with chronic low back pain. A pragmatic, randomized, single-blinded, controlled trial with 1-year follow-up. AB - STUDY DESIGN: A multicenter, randomized, single-blinded controlled trial with 1 year follow-up. OBJECTIVES: To evaluate the efficiency of progressively graded medical exercise therapy, conventional physiotherapy, and self-exercise by walking in patients with chronic low back pain. SUMMARY AND BACKGROUND DATA: Varieties of medical exercise therapy and conventional physiotherapy are considered to reduce symptoms, improve function, and decrease sickness absence, but this opinion is controversial. METHODS: Patients with chronic low back pain or radicular pain sick-listed for more than 8 weeks and less than 52 weeks (Sickness Certificate II) were included. The treatment lasted 3 months (36 treatments). Pain intensity, functional ability, patient satisfaction, return to work, number of days on sick leave, and costs were recorded. RESULTS: Of the 208 patients included in this study, 71 were randomly assigned to medical exercise therapy, 67 to conventional physiotherapy, and 70 to self-exercise. Thirty-three (15.8%) patients dropped out during the treatment period. No difference was observed between the medical exercise therapy and conventional physiotherapy groups, but both were significantly better than self-exercise group. Patient satisfaction was highest for medical exercise therapy. Return to work rates were equal for all 3 intervention groups at assessment 15 months after therapy was started, with 123 patients were back to work. In terms of costs for days on sick leave, the medical exercise therapy group saved 906,732 Norwegian Kroner (NOK) ($122,531.00), and the conventional physiotherapy group saved NOK 1,882,560 ($254,200.00), compared with the self-exercise group. CONCLUSIONS: The efficiency of medical exercise therapy and conventional physiotherapy is shown. Leaving patients with chronic low back pain untampered poses a risk of worsening the disability, resulting in longer periods of sick leave. PMID- 9854762 TI - Five-year follow-up study of a controlled clinical trial using light mobilization and an informative approach to low back pain. AB - STUDY DESIGN: A controlled clinical trial. OBJECTIVES: To examine the long-term effect of an informative approach to low back pain. SUMMARY OF BACKGROUND DATA: In management and prevention of low back pain, back school based on an ergonomic approach have played in important role. The effect of such informative interventions is not clear. METHODS: A 5-year follow-up study was done on patients included in a previous study. The outcome was measured by return to work or still on sick leave. The patients were allocated to an intervention group (n = 245) and a control group (n = 244). Only the intervention group was called in for examination and intervention and answered a battery of tests for psychological and health factors. The intervention apart from the clinical examination consisted of education in an "mini back school." The program was based on a new medical model for low back pain. RESULTS: Forty-seven (19%) of the patients in the intervention group, compared with 84 patients (34%) in the control group, were still on sick leave after 5 years (P < 0.001). There were fewer recurrences of sick leave (P < 0.03) in the intervention group than in the control group. Based on Internal Health Locus of Control, number of children, and income, 75% were correctly classified as nonreturners in the intervention group. CONCLUSIONS: This study indicates that subchronic low back pain may be managed successfully with an approach that includes clinical examination combined with information for patients about the nature of the problem, provided in a manner designed to reduce fear and give them reason to resume light activity. PMID- 9854763 TI - Range of motion and motion patterns in patients with low back pain before and after rehabilitation. AB - STUDY DESIGN: Data were collected from 27 patients who were participating in a rehabilitation program for chronic low back pain. The patients were tested on day 2 and day 11 of a 2-week rehabilitation program. OBJECTIVES: To determine specific characteristics of trunk motion associated with long-term dysfunction caused by low back pain of various origin, to determine if a neural network analysis system can be effective in distinguishing between patterns, and to determine if the rehabilitation has an effect on range and pattern of motion. SUMMARY OF BACKGROUND DATA: There is a lack of objective measures for evaluating the efficacy of rehabilitation programs. Numerous studies have established the difficulty of evaluating low back pain. Existing techniques, such as imaging methods, are in many cases either very rough and inaccurate or expensive and ineffective. A technique for evaluation of motion patterns in low back pain was developed based on analysis of dynamic motion features such as shape, velocity, and symmetry of movements. METHODS: Dynamic motion data were collected before and after rehabilitation from 27 patients with low back pain by using a triaxial goniometer. Range of motion and features of the movement, such as shape, velocity, and repetitiveness, were extracted for analysis. RESULTS: Motion features showed significant improvement after the rehabilitation program. CONCLUSIONS: A neural network based on kinematic data is an excellent model for classification of low back pain dysfunction. Such a system could markedly improve the management of low back pain for an individual patient. PMID- 9854764 TI - Lumbar olisthesis and lower back symptoms in elderly white women. The Study of Osteoporotic Fractures. AB - STUDY DESIGN: A Cross-sectional study. OBJECTIVES: To determine the prevalence of lumbar olisthesis among white women aged 65 years and older and its relation to low back pain and back function. BACKGROUND: Degenerative changes in the lumbar spine of elderly individuals may affect spinal stability, causing olisthesis (subluxation) of the lumbar spine. Neither the prevalence of this condition in the United States nor its relation to back symptoms has been studied previously. METHODS: Lateral radiographs of the lumbar spine for 788 women aged 65 years and older who were enrolled in the Study of Osteoporotic Fractures were digitized. Olisthesis (antero- and retro-) was assessed at L3-L4, L4-L5, and L5-S1. Back pain and function also were assessed. RESULTS: When olisthesis was defined as subluxation of 3 mm or more at any of the three levels studied, the overall prevalence of anterolisthesis was 29% and that of retrolisthesis was 14%. In 90% of women with anterolisthesis and 88% of women with retrolisthesis, subluxation occurred at a single vertebral level. The prevalence of anterolisthesis and retrolisthesis did not vary by smoking status, presence of diabetes, or history of oophorectomy. Anterolisthesis was not associated with the presence of back symptoms. Only retrolisthesis at L3-L4 was associated with a statistically significantly increased likelihood of lower back pain, greater severity of back pain, and impairment of back function. CONCLUSIONS: Anterolisthesis of 3 mm or more in the lower lumbar spine is relatively common among elderly women but is not correlated with back problems. Retrolisthesis at L3-L4 is associated with increased back pain and impaired back function. PMID- 9854767 TI - Updating the Veterinary Surgeons Act. PMID- 9854765 TI - Assessment of sagittal plane segmental motion in the lumbar spine. A comparison between distortion-compensated and stereophotogrammetric roentgen analysis. AB - STUDY DESIGN: Sagittal plane translatory and rotatory motion was measured in 15 lumbar motion segments of 8 patients by distortion-compensated and stereophotogrammetric Roentgen analysis. OBJECTIVE: To compare measurement precision of the new distortion-compensated Roentgen analysis protocol with that of the established Roentgen stereophotogrammetric technique under realistic clinical conditions. SUMMARY OF BACKGROUND DATA: Roentgen stereophotogrammetric analysis constitutes the most precise method available to assess segmental motion. Because of the invasive nature of the procedure, however, there is interest in alternative, noninvasive protocols suitable for studying larger patient cohorts. METHODS: In 8 patients, segmental motion of 15 lumbar segments that had undergone previous spinal surgery was assessed from stereo views by using Roentgen stereophotogrammetric analysis. Sagittal plane segmental motion was assessed by distortion-compensated Roentgen analysis. Sagittal plane translatory and rotatory motion data obtained by both methods were compared. RESULTS: With respect to Roentgen stereophotogrammetric analysis, sagittal plane rotation was determined by distortion-compensated Roentgen analysis with an error (standard deviation) of 1.4 degrees and a mean difference of less than 0.05 degree. Sagittal plane translation was determined by distortion-compensated Roentgen analysis, with an error of 1.25 mm and a mean difference 0.5 mm. CONCLUSION: Measurement precision of distortion-compensated Roentgen analysis is slightly inferior to that of Roentgen stereophotogrammetric analysis but substantially higher than that of conventional protocols assessing lumbar segmental motion. If measurement precision is considered adequate and if a noninvasive technique is indicated, distortion-compensated Roentgen analysis can be used to provide reliable motion data required for epidemiologic and clinical studies. PMID- 9854768 TI - Ambient temperature below which pigs should not be continuously showered in lairage. AB - Showering pigs with cold water during preslaughter lairage is thought to be useful in reducing the body temperature of hot, easily stressed animals. However, showering when the ambient temperature is too low could chill them too severely. To assess the effects of showering and to determine a temperature below which pigs should not be showered, pigs from one source, passing through a commercial slaughterhouse lairage, were split into two groups of approximately 50 each, showered and unshowered, on 10 days with a range of ambient temperatures. The pigs' behaviour and any damage to their skin were recorded, various measures of body temperature were taken before and after showering, and blood taken at slaughter was analysed for plasma creatine kinase, cortisol and lactate. Showering prevented the usual reduction in activity observed in pigs in lairage at high ambient temperatures. On the basis of the reduction in their flank temperature during showering, it is recommended that pigs should not be showered continuously if the temperature inside or outside the lairage falls below 5 degrees C, and showering should cease if they are seen to be shivering. PMID- 9854769 TI - Development of snake antivenom antibodies in chickens and their purification from yolk. AB - Adult white leghorn hens hyperimmunised with Brazilian snake venoms of the genus Bothrops and/or Crotalus produced antibodies capable of recognising, combining with and neutralising the toxic and lethal components of the venoms. The antibodies were first detected by an enzyme-linked immunosorbent assay two weeks after starting the immunisation schedule, reached the highest titres by the third week and remained high for at least 24 weeks. These antibodies are transferred to the egg yolk from which they were isolated as enriched IgY preparations by a combination of methods using positive and negative precipitation with sodium sulphate and/or caprylic acid. The yolk-derived IgY preparations contained antibodies which blocked the phospholipase A2-dependent haemolytic activity of both venoms and the haemorrhagic activity of Bothrops venom, and neutralised the toxic lethal activities of the venoms with good efficacy. The median effective dose (ED50) of the IgY anti-Bothrops venom was 592.5 microliters/2LD50 and, 1.0 ml neutralised 0.0675 mg of venom. The ED50 of the IgY anti-Crotalus venom was 457.5 microliters/3LD50 and 1.0 ml neutralised 0.075 mg of venom. PMID- 9854770 TI - Malignant schwannoma in a red deer (Cervus elaphus). AB - A five-year-old female red deer (Cervus elaphus) was in poor condition and severely lame on the left hindleg owing to a 19.4 cm x 15.9 cm mass involving and destroying the distal end (head) of metatarsal bones III and IV, the proximal sesamoid bones and the first phalanges (III and IV). The histopathological analysis revealed a spindle cell tumour with frequent palisade arrangement (Antoni type A pattern), and with highly anaplastic tumour cells in some areas. Structures resembling peripheral nerves were identified within the tumour. The neoplastic cells reacted with vimentin in a cytoplasmic pattern, and almost all of them reacted with S-100 protein in a nuclear and cytoplasmic pattern and did not express neurofilament, glial fibrillary acidic protein or keratins. This immunophenotype and the histopathological features were consistent with a diagnostic of malignant schwannoma. It was atypical because of the species affected, the location and the local malignancy. PMID- 9854771 TI - Prevalence of Lawsonia intracellularis in selected pig herds in Korea as determined by PCR. PMID- 9854773 TI - Incidence of oocyte nuclear maturation within the ovarian follicle of the bitch. PMID- 9854772 TI - Postoperative pain after ovariohysterectomy in the cat: a comparison of two anaesthetic regimens. PMID- 9854774 TI - Seroprevalence of antibodies to Bordetella bronchiseptica in cats in the Copenhagen area of Denmark. PMID- 9854775 TI - Coagulase-negative Staphylococcus intermedius isolated from milk from dairy cows in Israel. PMID- 9854776 TI - Pet insurance claim forms. PMID- 9854777 TI - Isolation of avian pneumovirus in QT-35 cells. PMID- 9854778 TI - [The use of computed tomography in stereotaxic operations in dyskinesia patients]. AB - Experience with 366 CT-stereotactic operations in varying movement disorders (Parkinson's disease, cerebral palsy, torsion dystonia, torticollis, hemihyperkinesis) is presented. Different deep structures, such as thalamic nuclei, subthalamic structures, palladium, putamen, caudate nucleus, dentate nucleus, and brachium conjunctivum or combination of these structures, were selected as a stereotactic target point. Anodic or radiofrequency stimulation, implantation of chronic intracerebral electrodes and neurotransplantation were used as methods of exposure. A procedure for CT-stereotactic determination of supratentorial and subtentorial target points is described. CT-guided stereotactic procedures in the treatment of dyskinesia are precise and less traumatic than ventriculography-guided procedures. PMID- 9854779 TI - [Functional stereotaxy with magnetic resonance tomographic guidance]. AB - The paper summarizes experience with stereotactic operations by magnetic resonance imaging data by using a Russian OREOL stereotactic manipulator. It describes methods of preoperative preparation of patients by employing diagnostic magnetic resonance tomographs. A number of points of the procedure for identification and localization of deep target structures from magnetic resonance images in parkinsonism, temporal epilepsy and some mental disorders. PMID- 9854780 TI - [The ultrastructural changes in the collagenous-elastic skeleton of the wall of the cerebral vessels in spasm in the acute period of a subarachnoid hemorrhage]. AB - Electron microscopic study of arteries in spasm and ischemia of the brain revealed substantial ultrastructural changes in the vascular wall. The changes are characterized by varying thickness, edema, dissociation of fibrillar and amorphous components of internal elastic membrane of the arterial wall, by lesion in the network of collagen fibers on its inner surface, by the presence of defibrotic and uncoiled collagenous fibers of the adventitia. The changes are followed by the impairment of the arterial skeleton and hence they lead to their altered elastic properties. PMID- 9854781 TI - [The postoperative monitoring of intracranial pressure in patients in the acute period of aneurysmatic hemorrhage]. AB - Changes in intracranial pressure (ICP) and cerebral circulation were evaluated from Doppler transcranial ultrasound (DTU) in 29 patients with spontaneous aneurysmal hemorrhage in acute rupture. There is a tendency for the pulse wave amplitude (PWA) of ICP to increase concurrently with higher ICP values (r = 0.768, p < 0.01), the tendency being smoother in patients with severe diffuse spasm (r = 0.573, p < 0.01). There was also a linear correlation between the ICP values and the pulse index of blood flow velocity (r = 0.783, p < 0.01). Evidence is given for the possibility of indirect assessment of ICP from DTU data and for that of indirect evaluation of cerebral perfusion from ICP monitoring data. In the latter case, the authors used a ICP PWA ratio. The findings suggest that the decrease in PWA/ICP values by less than 0.2 during intracranial hypertension and severe diffuse arterial spasm corresponds to the reduction in cerebral blood filling beyond the critical values. PMID- 9854782 TI - [The identification and functional tracking of the facial nerve during the removal of tumors of the cerebellopontine angle]. AB - Intracranial identification by electrostimulation and monitoring of the status of the facial nerve was intraoperatively used in 21 patients with cerebellopontine angle tumors of varying histological structure. Monopolar and bipolar electrostimulation, as well as electromyography and mechanography for recording the function of the facial nerve were compared. During removal of cerebellopontine angle tumors, identification and monitoring of the function of the facial nerve provide anatomic retention of this nerve when the tumor is radically eliminated. This is of essential significance for acoustic neurinomas. A combination of monopolar and bipolar stimulation for identification of the facial nerve simplifies removal of these tumors. As compared with electromyography, mechanography of facial nerve function during these operations is a more convenient technique due to the simplicity and absence of false operation for electric interferences. The study provided the optimum procedure for intracranial stimulation to identify the facial nerve within the cerebellopontine angle. PMID- 9854783 TI - [Variants in the growth of meningiomas of the base of the skull spreading into the orbit and paranasal sinuses]. AB - Meningiomas of the skull base involving the orbit and paranasal sinuses were diagnosed in 254 patients. The patients were operated on in 1982-1991. All craniocephalic meningiomas involve the anterior skull base bones. The involvement presents either as destruction (nodular tumor) or as hyperostosis (infiltrative growth of the tumor). All patients were divided into 8 groups with different location of skull bones involvement. A classification of meningiomas of skull base disseminating into the orbit and paranasal sinuses is proposed, proceeding from the anatomical principle. This classification helps plan a surgical access, scope of intervention, probable size of skull defect and method of its closure, possible postoperative functional and cosmetic defects. PMID- 9854784 TI - [Dopplerographic assessment of the autoregulation of the blood supply to the brain in neurosurgical pathology]. AB - The development of an informative, noninvasive technique for evaluating the reserve of the cerebral circulatory system is a topical task of clinical angioneurology. The authors developed methods of Doppler evaluation of an autoregulatory response from hyperemic changes after short-term regional hypotension caused by digital compression of the cervical carotid. They calculated the indices acceptable in routine clinical practice, defined the range of their values in health and in typical variants of cerebral circulatory insufficiency in patients with disseminated neurosurgical pathology. The carotid compression test by recording linear blood flow velocity in the cerebral arteries is a safe, valid, and reproducible method of semiquantitative assessment of the autoregulation reserve which may be used to determine the tension of resistive vessels in the middle cerebral arterial bed as an important index of the functional status of the cerebral circulatory system. PMID- 9854785 TI - [Craniocerebral trauma in children operated on earlier for hydrocephalus]. AB - The specific features of the clinical manifestations, diagnosis, management of brain injury are considered in 48 patients earlier anastomosed for progressive hydrocephalus. The leading features are mild focal neurological disorders, prevalent overall cerebral disturbances and no dislocational manifestations. These features are associated with the absence of hypertensive phenomena due to the presence of a drainage system and morphological substrate in children suffering from hydrocephalus, which affects the outcome of brain injury and its treatment. PMID- 9854786 TI - [A transmaxillary transnasal approach to tumors of the base of the skull]. AB - The microsurgical transmaxillary transnasal approach provides a wide access to the midline of the skull base. Since 1994 the approach was performed in 11 patients while resecting malignant and benign skull base tumors by involving the cribriform plate, orbit, maxillary and sphenoidal sinuses, nasal cavity, clivus, and cavernous sinus. The technique is suitable for extradural tumors of the skull base and can be combined with transcranial approaches for more extensive lesions. PMID- 9854787 TI - [The methodological bases of topographic neuropathomorphology]. AB - This paper describes the major features of topographic and anatomical studies of CNS structures, CSF circulation system, extracerebral structures and vertebral column. These methods include brain fixation, which is able to prevent post mortem brain deformation due to decalcification (even for block-preparations like "brain-skull base" or "posterior cerebral fossa-cervical spine"). The author also presents the following methods: spatial reconstruction of the brain and skull base subtracted from the series of slices of different thickness and various planes of transection; biopsy investigation of cerebral midline structures and CSF system in relation with skull base and convex structures; staged microdissection of tumors; filling of arterial and venous systems. All these modalities were used in examining the topographical features of craniopharyngiomas, hypophyseal adenomas, parasagittal and sagittal meningiomas, pineal tumors, VIII cranial nerve neurinomas, arachnoid cysts and in developing surgical approaches to the tumors of the third ventricle, skull base and foramen magna. PMID- 9854788 TI - [Epidemiological studies in neuro-oncology: the state of the art in Ukraine and abroad]. PMID- 9854789 TI - [Traumatic subarachnoid hemorrhages (the current aspects of the use of calcium blockers)]. PMID- 9854790 TI - Assembly in vitro of vitelline envelope components induced by a cortical alveolus sialoglycoprotein of eggs of the fish Tribolodon hakonensis. AB - Assembly in vitro of vitelline envelope (VE) components, which were precipitated by 50-70% saturated ammonium sulphate from VE extracts, was induced by the action of a sialoglycoprotein that is immunohistochemically localised in cortical alveoli of fish eggs and has serine proteinase activity. The VE components consisted of major bands of molecular mass about 150-120, 110-100, 70 and 27 kDa in addition to about 20 minor bands and contained a chorionic transglutaminase, visualised as two fluorescent bands by monodansylcadaverine staining. The VE component assembly in vitro was Ca(2+)-dependent, not induced if the sialoglycoprotein was pretreated with a serine proteinase inhibitor, and inhibited by the presence of p-chloromercuribenzoate, iodoacetamide or L-cysteine in the reaction medium system. Electron microscopy revealed that assembly in vitro of the VE components consisted of aggregates of network sheets, consisting of branching and anastomosing thin (approximately 27-52 nm) and thick (approximately 137-376 nm) filamentous substances. Separation by SDS-PAGE showed that a considerable number of VE components participated in the assembly in vitro in various amounts. These results suggest at least partial reproduction of the phenomena that occur in the process of fertilisation envelope (FE) formation, and provide a new approach to investigation of the process of FE assembly in vitro. PMID- 9854791 TI - Phosphorylation of endogenous and TEST-yolk buffer proteins by intact human sperm. AB - Protein kinase activity of intact, motile sperm was assessed by measuring the transfer of the terminal phosphate from [32P]ATP to tricholoroacetic acid (TCA) precipitable casein. The action of TEST (TES and Tris) yolk buffer (TYB) treatment on phosphorylation of sperm and TYB proteins was studied by detecting labelled phosphoproteins by autoradiography of polyacrylamide gel electrophoresis (PAGE). Results demonstrate that intact, forward-motile sperm have cell surface protein kinase activities. Although the difference between the kinase activity of freshly ejaculated sperm incubated in TYB was not significant, the protein phosphorylation during incubation in TYB showed that: (i) specific sperm surface proteins were phosphorylated to different degrees during the course of treatment; (ii) TYB proteins were phosphorylated by sperm during incubation; (iii) solubilised [32P]-labelled surface proteins were similar in molecular weight to TYB-labelled proteins. Taking into account that specific proteins on the human sperm surface undergo phosphorylation during incubation in TYB and that the sperm enzyme also acts specifically on some TYB proteins that become attached to the surface of the sperm, working hypotheses are proposed that suggest some correlation between the preservation of semen in TYB and the phosphorylation of proteins by intact human sperm. PMID- 9854792 TI - Fate of microinjected spermatid mitochondria in the mouse oocyte and embryo. AB - Mouse round spermatids labelled with MitoTracker were microinjected into Sr(2+) activated mouse oocytes. The labelled mitochondria were tracked up to the morula/blastocyst stage using fluorescence microscopy. The overall incidence of embryos with labelled mitochondria fell from 80% in the 1-cell zygote to 25% in 2 cell, 9% in 4-cell and approximately 1% in 8-cell or later stages. Thus it appears that almost all round spermatid mitochondria finally disappear from embryos during the 4-cell to 8-cell transition, as happens for mature spermatozoa (Cummins et al. Zygote 1997, 5:301-8). The spermatid mitochondria remained tightly bound together during this process. In contrast, labelled primary spermatocyte and cumulus mitochondria dispersed rapidly throughout the oocyte cytoplasm within 3 h. We hypothesise that spermatid mitochondria may be bound together by cytoskeletal elements produced in the early haploid spermatid. These elements, together with terminal differentiation of the sperm mitochondria, may be central to the processes by which the embryo 'recognises' the sperm mitochondria and inhibits inheritance of paternal mitochondrial DNA. These results suggest that round spermatid injection for infertile men will not pose a significant risk to offspring by transmitting abnormal mitochondrial genomes. PMID- 9854793 TI - LacZ transgene expression as a cell marker to analyse rescue from the 2-cell block in mouse aggregation chimeras. AB - Embryos from certain mouse strains are arrested at the 2-cell stage in cell culture ('2-cell block'), whereas those from other strains develop to the blastocyst stage under the same conditions. It was previously shown that blocking embryos can be rescued in culture by aggregation with an excess of 2-cell stages of a non-blocking strain such as CBA x C57BL/6 F2. Here we have employed a LacZ transgene in a blocking strain (NMRI) to follow the fate of rescued blastomeres up to the blastocyst stage. We found that rescued blastomeres can participate in both inner cell mass and trophoblast formation, thus completely overcoming the 2 cell block. PMID- 9854794 TI - Activation of embryonic genome in chick. AB - The earliest stages of development in most animals are under the control of maternally inherited information. The initiation of embryonic gene expression has been reported at the mid-blastula in amphibians and the mid-2-cell stage to the early morula in mammals. In chick embryos, embryonic gene expression was detectable at stage X (morula) and showed marked activation at stage XIII (blastula) with a gradual increase thereafter. Synthesis of rRNA and tRNA was low at stage X and was already the major class of RNA at stage XIII in chick embryos. The observed upregulation of RNA synthesis seems to coincide with a period of extensive fine structural differentiation when the first major cellular migrations start and signal the formation of the primitive streak in the chick embryo. PMID- 9854795 TI - Can alcohol retain the reproductive and genetic potential of sperm nuclei? Chromosome analysis of mouse spermatozoa stored in alcohol. AB - Alcohol is known to preserve genomic DNA and the primary structure of sperm protamines. To determine whether alcohol can retain the genetic and reproductive potential of mammalian sperm nuclei, mature mouse spermatozoa were stored in 70% ethanol or propanol for up to 2 months before injection into oocytes. Live offspring were obtained after injection of spermatozoa stored in 70% ethanol for 1 day at -20 degrees C. About 20% of the spermatozoa stored under this condition had normal chromosomes. The remaining 80% of spermatozoa and all the spermatozoa stored in 70% ethanol for 2 months had structurally aberrant chromosomes, and none could support the development of normal embryos. High concentrations of alcohol do not alter the primary structure of either DNA or small-molecular weight protamines. However, alcohol may modify protamine-protamine or protamine DNA interactions in a manner that results in the induction of DNA strand breaks during sperm chromatin decondensation within the oocyte. The limited success in obtaining normal offspring with ethanol-stored spermatozoa is encouraging. It may be possible to overcome these problems and develop a simple method for preserving mammalian spermatozoa without freezing. PMID- 9854796 TI - Dynamic changes of the microtubule system corresponding to the unequal and spiral cleavage modes in the embryo of the zebra mussel, Dreissena polymorpha (Mollusca, Bivalvia). AB - Unequal cleavage requires a highly organised cytoskeleton. We investigated the localisation of both tubulins and microtubular arrays in Dreissena eggs during and after fertilisation using confocal laser scanning microscopy. Freshly spawned eggs are arrested in metaphase I. A maternal pool of gamma-tubulin is found mainly in the centre of the asters of the meiotic spindle. The paternal pool of gamma-tubulin, present in the fertilising sperm, could not be traced within the egg, but a microtubule-organising centre forms near the male pronucleus at anaphase II. Male and female pronuclei grow as they migrate in the wake of their aster and rendezvous. First cleavage is unequal and starts without pronuclear fusion. At metaphase the two equal-sized asters span the entire egg in a symmetrical arrangement. At late metaphase the spindle shifts along its longitudinal axis into an eccentric position and the peripheral aster takes on an umbrella-like appearance, whereas the central aster remains spherical. The cleavage furrow becomes determined in the circumferential overlap of the asters. The inequality at second cleavage, however, is due to the unequal size of the asters. The third cleavage spindle also has asymmetrical asters and spindle shift was only observed in the D-cell. The spiral character is a result of an asymmetrical organisation of the larger, vegetal aster. Our results show that the arrangement of the gamma-tubulin clusters and of microtubules changes and develops during early development of Dreissena in a way that can explain the axis generating asymmetries in cell pattern and the spiral sense of cleavage. The major cytological characters expected to direct pattern formation in this phase of development are: size, position, and symmetry or asymmetry of both spindle and asters. PMID- 9854797 TI - Expression and subcellular distribution of rel/NF kappa B transcription factors in the preimplantation mouse embryo: novel kappa B binding activities in the blastocyst stage embryo. AB - We describe the expression patterns and subcellular localisation of murine rel/NF kappa B transcription factors and their inhibitors during preimplantation development in the mouse. We find that the known rel and I kappa B proteins are ubiquitously expressed during early murine embryogenesis. The differential persistence of individual rel and I kappa B transcripts in the first cleavage stage embryo and analysis of the 3'UTR suggests that rel and I kappa B protein expression may be regulated at the level of mRNA stability and translation. At the stages of early mouse embryogenesis examined, there was no evidence for strong induction of rel dimer translocation into the nucleus. However, novel kappa B-binding proteins are found in the mouse blastocyst and may provide an alternative mechanism for regulation of gene expression via the kappa B motif in the early mouse embryo. PMID- 9854798 TI - Time course of pronuclear formation and fertilisation after insemination in vitro and intracytoplasmic sperm injection of in vitro matured sheep oocytes. AB - The time course of sperm decondensation, oocyte activation, pronuclear formation and the possible causes of abnormalities after intracytoplasmic sperm injection (ICSI) and in vitro fertilisation (IVF) were examined. Frozen-thawed and pooled fresh semen from three different rams were washed and capacitated for ICSI or IVF. In vitro matured oocytes were cultured after sperm injection for 0.5, 0.75, 1, 2, 3, 4, 5, 6, 8, 18, 21 and 23 h, and oocytes were cultured after in vitro insemination for the same times other than 18 and 23 h. All oocytes were cultured in bicarbonate-buffered synthetic oviduct fluid medium (BSOF) supplemented with 2% oestrous sheep serum. A total of 746 metaphase II oocytes were injected with a single spermatozoon and 986 oocytes were inseminated for IVF. The earliest oocyte activation after ICSI was observed at 0.5 h, when 14.8% of oocytes were in anaphase II; this was earlier than after IVF, when only 6.4% of the oocytes exhibited anaphase II 1 h after insemination. Decondensing spermatozoa were first observed 1 h after ICSI and 3 h after insemination for IVF. The earliest female and male pronuclei after ICSI were observed at 2 and 3 h respectively, while the female and male pronuclei after IVF were observed at 4 h after insemination. The overall fertilisation rate was lower after ICSI (28.6%) than IVF (70.4%) but the percentage of abnormal fertilisation was not different between ICSI (8.7%) and IVF (15.2%). It was concluded that the fertilisation events were more advanced for ICSI than IVF, using injection and insemination time as reference points. The formation of male and female pronuclei were asynchronous after ICSI, in contrast to IVF when they appeared simultaneously at 4 h. Abnormalities found in fertilisation after ICSI may therefore be induced by the injection technique. PMID- 9854799 TI - Primary culture of porcine PGCs requires LIF and porcine membrane-bound stem cell factor. AB - We studied the effect of murine leukaemia inhibitory factor (LIF), human basic fibroblast growth factor (bFGF) and porcine stem cell factor (SCF) on the survival and/or proliferation of porcine primordial germ cells (PGCs) obtained from 27-day-old embryos in vitro. PGCs were cultured in embryonic stem cell (ESC) medium supplemented with or without either LIF (1000 IU/ml) alone or LIF together with bFGF (10 ng/ml). They were seeded on mitotically inactivated feeder cells, either STO or transfected STO cells (STO#8), expressing the membrane-bound form of porcine SCF. PGCs were identified by their alkaline phosphatase (AP) activity and counted after 1, 3 and 5 days in culture. After 1 day of culture, PGCs cultured on STO#8 cells showed significantly higher survival than PGCs cultured on STO cells (p < 0.05). The combined effect of SCF and LIF caused a significant increase in PGC number by day 3 of culture when PGCs were cultured on either STO cells (p < 0.01) or STO#8 (p < 0.001). When SCF and LIF were used together with bFGF no increase in the PGC number was observed. Our results suggest that the membrane-bound form of porcine SCF plays a pivotal role in the primary culture of porcine PGCs and that bFGF is not required in vitro. PMID- 9854800 TI - Protein kinase inhibitors induce the interphase transition by inactivating mitogen-activated protein kinase in mouse eggs. AB - The role of mitogen-activated protein (MAP) kinase in mouse egg activation induced by protein kinase inhibitors and a protein tyrosine kinase (PTK) inhibitor was investigated. Separated egg proteins were first probed with anti Active MAP kinase antibody and then re-probed with anti-ERK2 antibody. Staurosporine and Ro-31-8220, at concentrations that normally inhibit protein kinase C, did not affect egg activation or MAP kinase activity, while higher dosages caused egg activation. Staurosporine at 2 microM induced the metaphase interphase transition without emission of the second polar body (PB2), while Ro 31-8220 at 40 microM induced PB2 emission, first cleavage, and then the transition to interphase. Half the eggs were also activated by the PTK inhibitor genistein. In each treatment, the proportion of eggs that entered interphase was well correlated with the degree of MAP kinase inactivation. Artificial activation of this kinase by okadaic acid overcame the interphase transition. These data suggest that protein kinase inhibitors and a protein tyrosine kinase inhibitor induce the interphase transition by inactivating MAP kinase in mouse eggs. PMID- 9854801 TI - Covalently immobilized choline oxidase and cholinesterases on a methacrylate copolymer for disposable membrane biosensors. AB - Bienzymatic sensors for the determination of esters of choline were prepared by covalent co-immobilization of cholinesterases and choline oxidase on polymer membranes, obtained by radiation-induced copolymerization of 2-hydroxyethyl methacrylate and glycidyl methacrylate at low temperature. Optimization of the covalent attachment of choline oxidase and acetyl- or butyrylcholinesterase to copolymer was explored. The enzyme-modified polymers were applied on platinum electrodes to form amperometric sensors, based on the electrochemical detection of enzymatically developed hydrogen peroxide. Acetyl-, acetylthio-, butyryl-, and butyrylthiocholine contents in standard solutions were measured, and linear calibration curves were determined. Temperature and pH effects on the electrochemical response are described. PMID- 9854802 TI - Carob pod: a new substrate for citric acid production by Aspergillus niger. AB - The production of citric acid from carob pod extract by A. niger in surface fermentation was investigated. A maximum citric acid concentration (85.5 g/L), citric acid productivity (4.07 g/L/d), specific citric acid production rate (0.18 g/g/d), and specific sugar uptake rate (0.358 g/g/d) was achieved at an initial sugar concentration of 200 g/L, pH of 6.5, and a temperature of 30 degrees C. Other kinetic parameters, namely, citric acid yield, biomass yield, specific biomass production rate, and fermentation efficiency were maximum at pH 6.5, temperature 30 degrees C, and initial sugar concentration 100 g/L. The external addition of methanol into the carob pod extract at a concentration up to 4% (v/v) improved the production of citric acid. PMID- 9854803 TI - Preparing to react in the absence of uncertainty: I. New perspectives on simple reaction time. AB - Almost from the inception of psychological enquiry into reaction time (RT), a broad distinction has been drawn between two types of process-serving performance in RT tasks, namely, the on-line processes initiated by the arrival of the imperative signal and preparatory processes, which precede the imperative signal and may contribute to performance efficiency. Restriction of attention to on-line processing fosters the conclusion that the processes serving simple RT are no more than a subset of those involved in choice reactions. If, however, latencies can be reduced by specific preparation then the certitudes of the simple reaction task may invest it with distinctive properties. The possibility of preparation being sensory as well as motor needs to be considered. Until recently, attempts to study preparedness have failed to make controlled comparisons across simple and choice RT. However, recent work has shown that simple reactions have distinctive characteristics that set them apart from minimal choice reactions. The optimization of simple RT seems to depend upon attention-demanding processes which are probably preparatory in nature. When attention has to be shared with a concurrent task, much of the normal advantage of the simple reaction is lost. Neuropsychological studies suggest that patients may be found with abnormally extended simple RT but spared choice reactions. In these patients, the simple reactions are prolonged relative to those of controls but less vulnerable to imposition of a secondary task. Evidence is beginning to accrue to the effect that some distinctive processing features of simple reactions may be mediated by activity in the frontal lobes. PMID- 9854804 TI - Ageing and word processing competence: compensation or compilation? AB - Composition and compilation were examined as two alternative hypotheses relating to 'cognitive competence' in older adults. Techniques of molar equivalence molecular decomposition (ME-MD) and molecular equivalence-molar analysis (ME-MA) were used to investigate performance of experienced (over 100 hours) younger (18 30 years) and older (45-66 years) adults on a word processing task, requiring insertion and deletion of words from passages of text. There was some indication that typing skill may form the basis of a compensatory mechanism. However, differences between 'good' and 'poor' performers in the older sample supported the position that high levels of complex task skill in older adults are associated with the possession (possibly through maintenance) of high levels of basic cognitive capacities, and are not related to the integration of molecular task components. PMID- 9854805 TI - Computer anxiety: a comparison of pen-based personal digital assistants, conventional computer and paper assessment of mood and performance. AB - The recent growth of pen-based devices, such as the Personal Digital Assistant (PDA), offer mobility and a more natural interface than that of a conventional computer. The feasibility and application of the PDA for mood and cognitive assessment were investigated by examining possible interactions of individual characteristics and administration medium. Previous studies have provided evidence that individual characteristics of 'computer anxiety' and 'private self consciousness' divergently covaried with mood scores measured by computer and paper methods. To investigate the relationship between individual characteristics and medium effects, 136 paid participants were allocated to and completed mood assessment tasks and a short battery of cognitive tasks by either the computer, PDA or the paper method. Self-ratings of mood measured by these three modalities covaried divergently with measures of computer anxiety and private self consciousness. In addition, computer anxiety covaried with reaction time on the visual search task obtained on computers, but there was no such relationship when measured by a PDA. These results show that computer anxiety can affect the results of assessments of cognitive function as well as of mood ratings, and suggest that pen-based systems may have advantages over conventional computers in this respect. PMID- 9854806 TI - Gender differences in the dual-task effects on autobiographical memory retrieval during social problem solving. AB - A dual-task paradigm was used to explore the effects of cognitive load on social problem solving and autobiographical memory retrieval. The role that gender may play in mediating the relationship was also examined. Participants performed a secondary task concurrently with two primary tasks: (a) a cueing task, and (b) the Means-End Problem-Solving (MEPS) Task, during which they were required to attend to the memories retrieved during solution generation. Two dual-task conditions were employed in order that two levels of secondary task difficulty could be explored. Level of difficulty proved to be an important factor in the effects of resource reduction on the two primary tasks. Retrieval during the MEPS was effected by both the easy and difficult secondary task whereas retrieval on the cueing task was affected by the difficult task only. The results also showed that females (in contrast to males) favoured a more detailed SPS style using a specific memory database. Consequently, under central executive pressure, females' performance was significantly affected while males' performance remained largely unchanged. PMID- 9854807 TI - Personality traits and personality disorders. AB - The structure of personality disorder traits was examined in a sample of 400 undergraduates who completed the personality disorder questionnaire from the Structured Clinical Interview for DSM-III-R (SCID-II). The relations between personality disorder and normal personality traits indexed by the Eysenck Personality Questionnaire-Revised (EPQ-R) were examined. The three-cluster model of personality traits--as described in the DSM scheme--found equivocal support. Exploratory principal components analysis and confirmatory factor analysis found four broad factors of personality disorder that overlapped with normal personality traits: an asthenic factor related to neuroticism; an antisocial factor associated with psychoticism; an asocial factor linked to introversion extraversion; and an anankastic (obsessive-compulsive) factor. There is growing agreement about the number and type of broad personality disorder dimensions; similar dimensions may be found in clinical and non-clinical samples, suggesting that those people with personality disorders differ quantitatively rather than qualitatively from others; and there is substantial overlap between normal and abnormal personality dimensions. PMID- 9854808 TI - Perceptions of risk in motorcyclists: unrealistic optimism, relative realism and predictions of behaviour. AB - In the first phase of a prospective investigation, a national sample of motorcyclists completed a postal questionnaire about their perceptions of risk, their behaviour on the roads and their history of accidents and spills. In the second phase a year later, they reported on their accident history and behaviour over the preceding 12 months. A total of 723 respondents completed both questionnaires. Four sets of findings are reported. First, the group as a whole showed unrealistic optimism: on average, respondents believed themselves to be less at risk than other motorcyclists of an accident needing hospital treatment in the next year. Second, optimism was tempered by 'relative realism', in that respondents who were young and inexperienced saw themselves as more at risk than other motorcyclists, as did riders who reported risky behaviours on the road. Third, there was some evidence of debiasing by personal history, in that having a friend or a relative who had been killed or injured on the roads was associated with perceptions of absolute risk of injury or death--though there were no effects on comparative risk and no effects on any of the judgments of a history of accidents of one's own. Finally, there was good evidence that perceptions of risk predicted subsequent behaviour, though generally in the direction not of precaution adoption but of precaution abandonment: the greater the perceived risk at time 1, the more frequent the risky behaviour at time 2. The implications of the findings are discussed, and possible interpretations are suggested. PMID- 9854809 TI - Mono- and polyclonal antibodies as probes to study vitellin processing in embryos of the stick insect Carausius morosus. AB - During embryonic development, insect vitellins (Vt) are degraded by limited proteolysis to yield a number of lower-molecular weight polypeptides. The aim of the present study was to identify these polypeptides in the embryo and to verify how they relate to Vt polypeptides deposited in the oocyte during vitellogenesis. To this end a panel of poly- and monoclonal antibodies (Pab, Mab) was raised against Vt polypeptides and employed by immunoelectrophoresis and immunoblotting on embryos belonging to different developmental stages. Through this approach three major staining patterns were observed. First, Mab 4 reacts with both polypeptides B1 and E20, suggesting that polypeptide B1 is gradually trimmed to yield polypeptide E20 in late embryos. Second, Mab 12 is specific for polypeptide A3 which is retained unchanged throughout embryogenesis. Third, Pab anti-A2 and Mab 13 show that polypeptide A2 is processed to yield polypeptide E9 through limited proteolysis. In conclusion, the staining patterns reported in this study show that Vt polypeptides in developing embryos of the stick insect Carausius morosus undergo at least two major processing events concerning polypeptides B1 and A2. PMID- 9854810 TI - Purine metabolism in Echinococcus multilocularis. AB - The activities of the enzymes in Echinococcus multilocularis metacestodes involved in purine salvage were studied by HPLC. As in most parasites, this cestode relies entirely on salvage of preformed bases and nucleosides for its purine requirement. Therefore, these enzymes may be targets for drugs in the chemotherapeutic treatment of diseases caused by this parasite. The animals used in this study were gerbils (Meriones unguiculatus). Enzyme activities from sera and hepatic tissue in control and infected animals were similar with the exception of adenine phosphoribosyltransferase which showed an activity 4-fold greater in the serum from control than in serum from infected animals. In the parasite, adenine and hypoxanthine-guanine phosphoribosyltransferases and adenosine deaminase had the highest activities. Therefore, in E. multilocularis metacestodes, this pathway seems to be important for the parasite's metabolism. PMID- 9854811 TI - Comparative studies of suidatrestin, a specific inhibitor of trehalases. AB - Suidatrestin, isolated from a Streptomyces strain, was characterized as a new trehalase inhibitor. Its inhibitory potential was 7 to 50-fold higher than that of validamycin when tested against insect, fungal and mammalian trehalases. The kinetic properties of suidatrestin were studied in vitro with trehalases from flight muscle mitochondria of the fly, Protophormia terraenovae, from larval midgut of the moth, Spodoptera littoralis, and from porcine kidney, as well as with maltase from yeast. Suidatrestin was inactive on maltase but inhibited all trehalases with IC50 values of 0.08-0.1 microM; Ki values ranged from 0.02 to 0.05 microM. The very low Ki/K(m) ratios (3.9 x 10(-6) -4.9 x 10(-6)) indicated excellent in vitro inhibitory action of suidatrestin. When injected into larvae of S. littoralis, suidatrestin required high and repetitive doses which lead to reversible inhibition of larval growth only. Consecutive omission of the inhibitor even stimulated weight increase above that of controls. Significant mortality was achieved at a rather high dose only. Injection of a growth inhibiting dose of suidatrestin did not change hemolymph osmolality as a measure of sugar concentration. The discrepancy between in vitro and in vivo potency of suidatrestin may be understood once its chemical structure is fully known. PMID- 9854812 TI - Studies on porcine high molecular weight kininogen. An improved method for the purification of porcine high molecular weight kininogen and cleavage of the kininogen by the action of porcine plasma kallikrein. AB - By introduction of stepwise DEAE Sephadex A-50 and copper-Chelating Sepharose 6B column chromatographies, about 18.5 mg of high molecular weight kininogen (HK) composed of a single polypeptide chain was obtained from 500 ml of porcine plasma. Molecular weights of reduced or non-reduced preparation were estimated to be 110 kDa and 116 kDa, respectively, by SDS-PAGE. Using the preparation, cleavage of HK by porcine plasma kallikrein (KK) was investigated. A single polypeptide HK was cleaved into two chains cross-linked by disulfide bond(s), accompanying the release of kinin. Further degradation was not observed. Molecular weights of heavy-chain (H-chain) and light-chain (L-chain) were estimated to be 61 kDa and 56 kDa, respectively, by SDS-PAGE. The amino- (N-) terminal sequences of intact HK, reduced and carboxymethylated- (RCM-) H-chain, RCM-L-chain and the peptide around the kinin moiety obtained by BrCN digestion were determined. Their sequences were highly homologous with those of bovine or human HK. These results indicate that plasma KK first cleaved the Arg-Ser bond of HK, and formed nicked HK. The second cleavage yielded bradykinin (BK) and kinin free protein, which was apparently of equal size to the nicked HK. The structure of HK was from the N-terminus to the carboxy- (C-) terminus, H-chain-BK-L-chain. PMID- 9854813 TI - Production of a biologically active novel goldfish growth hormone in Escherichia coli. AB - Goldfish pituitary contains two types of growth hormones. One with five cysteine residues (type-I) similar to other Cyprinid GHs, and the other with four Cys residues (type-II) similar to those of other fish and tertapod species. Recombinant goldfish type II GH (gfGH-II) was produced in Escherichia coli using the pRSETB expression vector. The gfGH-II was produced fused to a leader sequence, which sequestered into inclusion bodies after expression. The inclusion bodies were solubilized using sodium hydroxide and the fusion protein purified by chelating affinity chromatography. Subsequently, gfGH-II was cleaved and analyzed by Western blotting, using a specific antiserum. For comparison we also produced recombinant common carp GH (cGH) which has 95% similarity to gfGH-II, and tested their growth promoting activity in goldfish. Both forms of GH significantly increased the growth rate of goldfish (P < 0.05), although cGH was found to have a somewhat higher potency than gfGH-II. PMID- 9854814 TI - Role of isozyme group-specific sequence 4 in the isozyme-specific properties of human aldolase C. AB - To assess which regions of the aldolase C molecule are required for exhibiting isozyme-specific kinetic properties, we have constructed nine chimeric enzymes of human aldolases A and C. Kinetic studies of these chimeric enzymes revealed that aldolase C absolutely required its own isozyme group-specific sequences (IGS), particularly IGS-4, for exhibiting the characteristics of aldolase C which differ significantly from those of isozymes A and B (Kusakabe T, Motoki K, Hori K. Human aldolase C: characterization of the recombinant enzyme expressed in Escherichia coli. J Biochem (Tokyo) 1994;115:1172-7). Whereas human aldolases A and B required their own isozyme group-specific sequences-1 and -4 (IGS-1 and -4) as the main determinants of isozyme-specific kinetic properties (Motoki K, Kitajima Y, Hori K. Isozyme-specific modules on human aldolase A molecule. J Biol Chem 1993;268:1677-83; Kusakabe T, Motoki K, Sugimoto Y, Takasaki Y, Hori K. Human aldolase B: liver-specific properties of the isoenzyme depend on type B isozyme group-specific sequence. Prot. Eng. 1994;7:1387-93), the present studies indicate that the IGS-1 is principally substitutable between aldolases A and C. The kinetic data also suggests that the connector-2 (amino acid residues 243-306) may modulate the interaction of IGS units with the alpha/beta barrel of the aldolase molecule. PMID- 9854815 TI - Identification and characterization of the tumor suppressor p53 in channel catfish (Ictalurus punctatus). AB - Herein is presented the sequence of a catfish full-length p53 cDNA obtained from a cloned B cell line cDNA library. Southern blot analyses determined that a restriction fragment linked polymorphism (RFLP) existed with PstI among outbred catfish. Western blot analyses demonstrated that, when compared to PBLs, the catfish leukocyte lines express higher levels of p53 protein. Additionally, the results of Western blot analyses and in vitro translation experiments suggest that the catfish leukocyte lines may produce truncated forms of p53 due to internal initiation. PMID- 9854816 TI - Phosphorylation of rod outer segment proteins modulates phosphatidylethanolamine N-methyltransferase and phospholipase A2 activities in photoreceptor membranes. AB - The activities of enzymes involved in lipid metabolism--phospholipase A2 (PLA2) and phosphatidylethanolamine N-methyltransferase (PE N-MTase)--were found to be differently affected by pre-incubation of rod outer segments (ROS) under protein phosphorylating or dephosphorylating conditions. Exposure to cAMP-dependent protein kinase (PKA), under dark or light conditions, produced a significant increase in PE N-MTase activity, whereas PLA2 activity decreased. Under standard protein kinase C (PKC) phosphorylating conditions in light, PE N-MTase activity was stimulated and PLA2 activity was not affected. When the assays were performed in the dark, both enzymatic activities were unaffected when compared to the corresponding controls. Incubation of ROS membranes in light in the presence of PKC activators phorbol 12,13-dibutyrate (PDBu) and dioctanoylglycerol (DOG) resulted in the same pattern of changes in enzyme activities as described for standard PKC phosphorylating condition. Pre-incubation of membranes with the PKC inhibitor H-7 reduced the stimulation of PDBu on PE N-MTase activity, and had no effect on PLA2 activity in ROS membranes incubated with the phorbol ester. Pre treatment of isolated ROS with alkaline phosphatase resulted in decreased PE N MTase activity and produced a significant stimulation of PLA2 activity under dark as well as under light conditions when compared to the corresponding controls. These findings suggest that ROS protein phosphorylation and dephosphorylation modulates PE N-MTase and PLA2 activities in isolated ROS, and that these activities are independently and specifically modulated by particular kinases. Furthermore, dephosphorylation of ROS proteins has the opposite effect to that produced by protein phosphorylation on the enzymes studied. PMID- 9854817 TI - Coevolution of actin and associated proteins: an alpha-actinin-like protein in a cyanobacterium (Spirulina platensis). AB - Actin, together with associated proteins, such as myosin, cross-linking or capping proteins, has been observed in all eukaryotic cells. Presence of actin or actin-like proteins has also been reported in prokaryotic organisms belonging to the cyanobacteria. Our aim was first to extend the characterization of an actin like protein to another prokaryotic cell, i.e. Spirulina, then to compare the antigenic reactivity of this new protein with that of Synechocystis and skeletal actins. We observed that some of the conserved antigenic epitopes corresponded to actin regions known to interact with cross-linking proteins. We also report for the first time that alpha-actinin and filamin purified from chicken gizzard both interact with a prokaryotic actin-like protein. Finally, we searched for the occurrence of a cross-linking protein in these cyanobacteria and identified a 105 kDa protein as an alpha-actinin-like protein using specific antibodies. PMID- 9854818 TI - Characterization of a Schistosoma mansoni homologue of the gene encoding the breast basic conserved protein 1/L13 ribosomal protein. AB - The Schistosoma mansoni gene sequence encoding the breast basic conserved protein 1/ribosomal protein L13 has been isolated from an adult worm cDNA library using the Expressed Sequence Tag strategy. The cDNA codes for a putative protein of 184 amino acids which is approximately 55% identical to other eukaryotic L13 ribosomal proteins. A PCR amplified genomic fragment containing the coding region of the gene was seen to possess only a single large intron interrupting the open reading frame. Studies of gene expression by RT-PCR showed the transcript is expressed in distinct stages of the parasite life cycle. The cDNA was also hybridized with an ordered cosmid library of S. mansoni and the identified cosmids were mapped to chromosomes 3 and W by chromosomal in situ suppression hybridization. PMID- 9854819 TI - Purification, characterisation and distribution of ovine neuronal nitric oxide synthase. AB - Nitric oxide (NO) is an ubiquitous intercellular messenger molecule synthesised from the amino acid L-arginine by the enzyme nitric oxide synthase (NOS). A number of NOS iso-enzymes have been identified, varying in molecular size, tissue distribution and possible biological role. To further understand the role of NO in the regulation of neuroendocrine function in the sheep, we have purified and characterised ovine neuronal NOS (nNOS) using anion exchange, affinity and size exclusion chromatography. SDS-PAGE reveals that ovine nNOS has an apparent denatured molecular weight of 150 kDa which correlates well with the other purified nNOS forms such as rat, bovine and porcine. The native molecular weight predicted by size-exclusion chromatography was 200 kD which is in close agreement with that found for porcine and rat nNOS. Internal amino acid sequences generated from tryptic digests of the purified ovine nNOS are highly homologous to rat nNOS. There was no significant difference in the cofactor dependence and kinetic characteristics of ovine nNOS when compared to rat and bovine nNOS, (K(m) for L arginine 2.8, 2.0 and 2.3 microM respectively). A polyclonal anti-peptide antibody directed toward the C-terminal end of the rat nNOS sequence showed full cross-reactivity with the purified ovine nNOS. Immunohistochemical and Western analysis using this antiserum demonstrate the expression of nNOS in the cortex, cerebellum, hypothalamus and pituitary of the sheep. The lack of staining in the neural and anterior lobes of the pituitary seems to suggest that NOS plays a varied role in the control of endocrine systems between species. PMID- 9854820 TI - Isoforms of an N-acetyl-beta-D-glucosaminidase from the Antarctic krill, Euphausia superba: purification and antibody production. AB - Two forms of the chitinolytic enzyme N-acetyl-beta-D-glucosaminidase (NAGase, EC 3.2.1.52) have been isolated from the Antarctic krill, Euphausia superba, in order to study their potential role in temperature adaptation processes. A chromatographic protocol was developed that allowed complete separation of the two enzyme forms, named NAGase B and NAGase C. The latter was purified to homogeneity with 600-fold enrichment and a yield of 17%. The molecular mass was 150 kDa. NAGase B showed characteristics of a glycoprotein due to affinity towards concanavalin A sepharose, while NAGase C did not. Highly specific polyclonal antibodies to NAGase C [anti-(E. superba-NAGase C)-IgG] showed only negligible cross-reactivity with NAGase B isoforms. A comparison with the Northern krill, Meganyctiphanes norvegica, revealed a corresponding chromatographic pattern with two main activity peaks, for differentiation named NAGase II and NAGase III. Application of the antibody on M. norvegica revealed a high specificity toward NAGase III and a low cross-reactivity with NAGase II. First indication is given that the two forms are no isoenzymes in a strict sense but instead may have different functions in the metabolism of krill. PMID- 9854821 TI - Enzymes related to catecholamine biosynthesis in Tetrahymena pyriformis. Presence of GTP cyclohydrolase I. AB - We first identified GTP cyclohydrolase I activity (EC 3.5.4.16) in the ciliated protozoa, Tetrahymena pyriformis. The Vmax value of the enzyme in the cellular extract of T. pyriformis was 255 pmol mg-1 protein h-1. Michaelis-Menten kinetics indicated a positive cooperative binding of GTP to the enzyme. The GTP concentration producing half-maximal velocity was 0.8 mM. By high-performance liquid chromatography (HPLC) with fluorescence detection, a major peak corresponding to D-monapterin (2-amino-4-hydroxy-6-[(1'R,2'R)-1',2',3' trihydroxypropyl]pteridin e, D-threo-neopterin) and minor peaks of D-erythro neopterin and L-erythro-biopterin were found to be present in the cellular extract of Tetrahymena. Thus, it is strongly suggested that Tetrahymena converts GTP into unconjugated pteridine derivatives. In this study, dopamine was detected as the major catecholamine, while neither epinephrine nor norepinephrine was identified. Indeed, this protozoa was shown to possess the activity of a dopamine synthesizing enzyme, aromatic L-amino acid decarboxylase. On the other hand, activities of tyrosine hydroxylase or tyrosinase which converts tyrosine into dopa, the substrate of aromatic L-amino acid decarboxylase, could not be detected in this protozoa. Furthermore, neither dopamine beta-hydroxylase activity nor phenylethanolamine N-methyltransferase activity could be identified by the HPLC methods. PMID- 9854822 TI - Isolation and characterization of calmodulin in the pheromone gland of the silkworm, Bombyx mori. AB - Production of the sex pheromone bombykol in the silkworm, Bombyx mori, is regulated by a neurohormone termed pheromone biosynthesis activating neuropeptide (PBAN). It has been suggested that the external signal of PBAN in this species is transmitted to the intracellular cascade reactions consisting of Ca2+/calmodulin (CaM) complex and phosphoprotein phosphatase. To demonstrate the molecular mechanisms regulated by PBAN, we attempted to characterize CaM in the pheromone gland of B. mori. By using ion-exchange and RP-HPLC, B. mori CaM was purified from the cytosolic fraction of the pheromone gland. The primary structure was deduced by composition/sequence analysis and mass spectrometric analysis of the fragment peptides obtained from enzymatic and chemical fragmentations. The amino acid sequence of B. mori CaM was identical with Drosophila CaM deduced from the CaM gene of D. melanogaster, suggesting that insects have well conserved the molecule of CaM. PMID- 9854823 TI - The unique lipid composition of gecko (Gekko Gekko) photoreceptor outer segment membranes. AB - This study investigated the lipid and fatty acid composition of gecko photoreceptor outer segment membranes which contain the P521 cone-type pigment. The lipids of gecko photoreceptor outer segment membranes were first extracted and separated by thin layer chromatography (TLC) and then analyzed by gas chromatography (GC). Our results show that gecko photoreceptor outer segment membranes contain less phosphatidylethanolamine (PE) and more phosphatidylcholine (PC) and phosphatidylserine (PS) compared with those of bovine and frog. The content of the polyunsaturated fatty acid, docosahexaenoic acid (DHA), in PC and PS is also the highest yet reported (55 and 63%, respectively). These lipid differences may provide some insight into the specific lipid requirements of cone type pigments. PMID- 9854824 TI - Bothrops jararaca snakes produce several bothrojaracin isoforms following an individual pattern. AB - More than one isoform of bothrojaracin (BJC), a potent and specific thrombin inhibitor isolated from Bothrops jararaca venom, has been found in individual venoms collected from adult snakes. Variations in snake venom composition have previously been associated with factors such as age, sex, geographic origin, season of the year and diet. In order to obtain further information concerning individual patterns of expression of BJC isoforms, we have analyzed five individual Bothrops jararaca snake venoms collected at the same time from adult female snakes from the same geographic region. As expected, crude venoms showed a similar migration pattern on SDS-PAGE. BJC was purified using a procedure which includes an affinity chromatography step (PPACK-thrombin Sepharose). A slight variation in the amount of BJC obtained from individual venom samples was noticed. Inhibition of thrombin-induced platelet aggregation as well as migration pattern on SDS-PAGE (under reducing and non-reducing conditions) and isoelectric focusing varied considerably among BJC samples from the five snakes. The amino terminal sequences (residues 1-34) of individual BJC samples were compared with the sequence deduced from isolated cDNAs encoding alpha and beta chains of BJC. A high degree of homology was detected, although some residues differed from one sample to other. Altogether, data confirmed the heterogeneity found for BJC purified from individual snakes. Thus, the results indicate that: (1) individual specimens of Bothrops jararaca have different patterns of BJC isoform expression; and (2) it seems that genetic factors, at least in part, determine the variability found in BJC production. PMID- 9854825 TI - The death of King Charles XII--the forensic verdict. AB - King Charles XII of Sweden was killed in 1718 during his siege of the Danish fortress of Fredriksten. For 276 years, it remained an open question whether the lethal bullet came from the enemy or from a Swedish assassin. Now, a treatise published by a Swedish historian finally proves that the King's death was a case of political murder. Ballistic circumstances and the Danish ammunition then available are incompatible with a random shot from enemy quarters. Major-general Carl Cronstedt possessed the expertise needed to make an assassination look like a war casualty. It appears that the King was shot with a makeshift jacketed bullet long before jacketed bullets came into common use. PMID- 9854826 TI - Sudden death in cases with anomalous origin of the left coronary artery. AB - Ectopic origin of the left coronary artery from the right sinus of valsalva with anterior and posterior courses are thought to be benign anomalies. CASE REPORTS: a 58-year-old woman died suddenly after a car accident without having sustained any injuries. The only abnormal finding was an ectopic origin of the left coronary artery from the right sinus with anterior free wall course. The anomaly was complicated by the absence of a left descending branch and a hypoplastic circumflex artery. A 38-year-old male died suddenly during work. Beside the ostium of the right coronary artery originated the normal calibered circumflex branch of the left coronary artery which passed behind the aorta. The left anterior descending branch had a normal origin. There was a 2 cm in diameter transmural infarct observed microscopically. PMID- 9854827 TI - Fatal gastrointestinal haemorrhage due to coexisting primary aorto-enteric and aorto-colic fistulae. Complicating untreated atheromatous abdominal aortic aneurysm. AB - We report a case of aorto-enteric and aorto-colonic fistulae in a 73 year old man with no previous abdominal surgery which resulted in massive gastrointestinal haemorrhage and death and was undiagnosed despite several hospital admissions and endoscopic examinations. This case serves to highlight an extremely rare but fatal condition, which is potentially treatable, if an early diagnosis is made. PMID- 9854828 TI - A preliminary investigation into postmortem changes in skinfold impedance during the early postmortem period in rats. AB - A highly accurate and reproducible technique was used to measure the electrical impedance (1 kHz) of a lateral abdominal skinfold, in each of eight rat cadavers stored at 9.0 +/- 0.7 degrees C, during the early postmortem period. Impedance increased exponentially between 1 and 120 h post mortem. Whether or not similar changes occur during the human postmortem period, and their potential value as a means of objective estimation of postmortem interval (PMI), remains to be established. PMID- 9854829 TI - Alzheimer changes are common in aged drivers killed in single car crashes and at intersections. AB - With increasing age, diseases affecting the cognitive functions are more frequent. These diseases may increase the risk for fatal car crashes. We analyzed the frequency of neuropathological alterations characteristic of Alzheimer's disease (i.e. neuritic and diffuse plaques, and neurofibrillary tangles) in two association areas of the brain, parietal and frontal cerebral cortex, from 98 fatally injured aged drivers. In the age groups of 65-75 and over 75 years of age, 50% and 72% of the drivers, respectively, had neuritic plaques in either parietal and/or frontal cortex. In 14% of all killed drivers the number of neuritic plaques reached the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) age-related histologic score C, which indicates the diagnosis of Alzheimer's disease (AD), and an additional 33% had score B, which suggests the diagnosis of AD. Neuropathological AD changes were most common in the brains of drivers killed in single vehicle crashes, followed by multivehicle crashes at intersections and least common in multivehicle crashes elsewhere, but the differences did not reach statistical significance. In a great majority (80-85%) of cases the killed aged driver was the guilty party of the crash. The results imply, that incipient AD may contribute to fatal crashes of aged drivers, and therefore the forensic autopsy of these victims should include neuropathological examination. PMID- 9854830 TI - Anatomical reconstruction of fragments of burned human bones: a necessary means for forensic identification. AB - The aim of any forensic inquiry concerning accidental or criminal cremations is to identify the victims. This identification depends on an essential approach: a careful collection and ultrasonic cleaning of the fragments and splinters of burned bones, and reconstruction--as far as possible--of bone specimens, which implies that forensic anthropologists are well experienced in the field of burned bones. PMID- 9854831 TI - The prevalence of dextropropoxyphene in autopsy blood samples. AB - The prevalence of dextroproxyphene (DXP) in the total medico-legal autopsy material in Sweden during 1992 to 1996 was examined. Simultaneous findings of paracetamol and alcohol in the blood were considered in the analyses. DXP in peripheral blood was found in 1782 (7.5%) of the 23,691 cases analysed during 1992-1996. The autopsy prevalence of DXP increased by 25% from 1992 to 1996. The mean blood DXP concentration was 1.62 micrograms/g (the blood level of DXP after a therapeutic dose is 0.05-0.75 microgram/g). The blood DXP level was < 0.75 microgram/g in 947 cases and > or = 0.75 microgram/g in 835 cases. The cases < 50 years of age had a significantly higher mean concentration (2.36 micrograms/g) than those > or = 50 years (1.04 micrograms/g). Paracetamol in the blood was found in 53% of the DXP cases (mean 75.0 micrograms/g; therapeutic level 2.5-25 micrograms/g) and alcohol in 43% (mean level 0.14%). According to the death certificates 54% (956) died from fatal poisoning. Among these, 74% (707) showed a blood DXP concentration > or = 0.75 microgram/g. Other Scandinavian countries, Denmark and Norway have reduced the rate of fatal DXP poisonings through government regulations for prescription. As the defined daily dose/1000 inhabitants during a 12-month period (DDD) of DXP preparations in Sweden (14.4 in 1996) is six times as high as in Denmark and nine times as high as in Norway, introduction of similar regulations in Sweden should be considered. PMID- 9854832 TI - Nuclear morphometry of the myocardial cells as a diagnostic tool in cases of sudden death due to coronary thrombosis. AB - Sudden cardiac death due to underlying coronary artery thrombosis is one of the leading causes of death. However, in a significant percentage of individuals who died suddenly, no indication of myocardial infarction is found during post-mortem examination, especially when the time interval between appearance of symptoms and death is short. In the present study, we have evaluated certain nuclear morphometric parameters, such as, minimum, maximum, mean and standard deviation of perimeter and area in 20 individuals who died of coronary artery thrombosis, within 1 h from symptoms onset. Furthermore, the above parameters were compared with those of a control population of 20 individuals whose sudden death was caused by traffic accidents. Statistical elaboration of the results by means of t test, Mann-Whitney (U-test) and analysis of covariance (adjusting for age), showed a statistically significant difference for all variables except for the minimum area. With stepwise discriminant analysis method, the mean perimeter was selected as the best predictor of cardiac death. Mean perimeter achieved a correct reclassification percentage (based on Fisher's linear discriminant function) of 92.5% (85% and 100% for cases and controls, respectively). Moreover, by applying the cut-off of 172 microns, we could identify the individuals who died suddenly because of coronary artery thrombosis with a specificity of 100% (sensitivity 85%, P < 0.001). Our results show that nuclear morphometry of the myocardial cells is a reliable diagnostic tool for the diagnosis of coronary thrombosis based lesion in cases of sudden death, even when methods trying to verify the presence of infarction fail to do so. PMID- 9854834 TI - Morphological changes in traumatized skeletal muscle: the appearance of 'opaque fibers' of cervical muscles as evidence of compression of the neck. AB - Cervical muscles of 15 cases of compression of the neck and the other traumatized skeletal muscles from 54 autopsy cases were examined histologically and immunohistochemically. Round and thick fibers, that is, opaque fibers were observed in the muscles beneath the compression marks on the neck, whereas in areas where no force had been applied, such fibers did not exist. Furthermore, opaque changes appeared around cavities, which formed within severely compressed injured muscle tissue. The pattern of appearance of opaque fibers in the cases of compression of the neck were mainly divided into two types, 'diffuse' pattern and 'focal' pattern. In both blunt and sharp force injuries, the production of opaque fibers were common in the area directly where damage had been inflicted. In stab wounds especially, the appearance of opaque fibers and wavelike fibers, which fanned out and showed a different color from the other areas, was limited to the marginal zone. These results suggest that there is a close relationship between opaque changes and force. Opaque fibers are probably produced by damage to muscles due to extremely applied force. The high incidence of opaque fibers beneath compression marks on the neck should be regarded as a useful indication of strangulation and/or hanging. Furthermore, the distribution and direction of force to the neck might be presumed by the pattern of opaque fibers in cervical muscles, to a certain extent. PMID- 9854833 TI - The manner of death among fatalities where dextropropoxyphene caused or contributed to death. AB - Dextropropoxyphene (DXP) is one of the most prescribed analgesic compounds in Sweden. To investigate the manner of death among fatalities where DXP caused or contributed to death all medico-legal autopsies performed in Sweden in 1992-1996 were analysed on the bases of toxicological analyses and death certificates. DXP in peripheral blood was found in 1782 (7.5%) of the total 23,691 blood samples. According to the death certificates 956 (54%) of the 1782 cases were classified as fatal DXP poisoning. Among these, the manner of death was classified as accidental in 49 cases (5%), suicidal in 542 cases (57%) and undetermined in 365 cases (38%). The reported manner of death differed between the six forensic medicine districts in Sweden. The accident rate differed significantly between the district with the highest rate (9%) and the districts with the lowest rate (1%). One district had a significantly higher incidence of suicide (73%) than four of the other districts, while another district had a significantly lower incidence of suicide (33%) than all the other districts. The accident classification rate among the physicians performing ten or more autopsies varied from 0% to 17%, the suicide classification rate from 25% to 83% and the rate of undetermined manner of death from 8% to 71%. A major conclusion drawn from this study is that accidental DXP fatalities may be underestimated. This may have serious consequences, as under-reporting of accidental DXP fatalities will increase the risk that knowledge of the high toxicity of DXP will not reach the population consuming this drug. Since valid death statistics concerning the manner of death at DXP fatalities are needed to provide the base for preventive actions, special attention should be paid to the classification process, in order to increase the uniformity of the assessments among the different physicians, and to avoid under-reporting of accidents. PMID- 9854835 TI - Bruising in non-accidental head injured children; a retrospective study of the prevalence, distribution and pathological associations in 24 cases. AB - Non-accidental head injury, be it shaking, impact(s) or a combination of the two, is characterised by subdural and/or subarachnoid haemorrhages with retinal haemorrhages, but minimal or absent external cranio-facial trauma. The classical assault scenario depicts the infant being gripped around the head, face, chest and abdomen and shaken or being gripped by a limb and swung. This gripping might be expected to leave physical evidence in the form of bruising. A study was undertaken to establish the prevalence, distribution and pathological association of external bruising in 24 cases of fatal non-accidental head injury in children. At autopsy, 17 cases had new external bruises, 15 old external bruises and 13, a combination of both. However, seven (29%) cases showed no fresh external bruising and five (21%) showed no external bruising at all. Thus, external bruising may be absent in children with fatal intracranial injury. The face was shown to be the commonest site of bruising followed by the forehead and buttocks. Limb, chest and abdominal bruising were found to be uncommon. Retinal haemorrhages were confirmed in 23 (96%) cases. It is hypothesised that bruising, when present, may be a result of abuse in the form of punches and slaps rather than due to gripping during the assault. We discuss why gripping does not necessarily result in external bruising. PMID- 9854836 TI - High cervical stress and apnoea. AB - The aim of this study was to investigate vegetative reactions in infants after mechanical irritation of the suboccipital region. The investigation is based on 199 infants who were observed while being treated with a suboccipital impulse (manual therapy). The results revealed vegetative reactions in more than half of all cases (52.8%, n = 105). The frequency of such vegetative reactions observed was at follows: flush 48.7% (n = 97), apnoea 22.1% (n = 44), hyperextension 13% (n = 26), and sweating 7.5% (n = 15). It is pointed out that approximately 25% of all the infants examined reacted by apnoea due to a mechanical irritation of the suboccipital region. This symptom was part of an extensive vegetative reaction. This method of inducing an apnoea has not yet been described; from this it follows that there are close relations to sudden infant death. PMID- 9854837 TI - The use of vertebral ring epiphyseal union for age estimation in two cases of unknown identity. AB - The objective of this case study was to demonstrate the utility of thoracic and first two lumbar vertebral ring epiphyseal union as an accurate and reliable method of age estimation for teenage and young adult skeletons. Vertebral ring epiphyseal union was used as a method of age estimation in two cases of osteological analysis for individuals of unknown identity. Case 1 involved male skeletal remains where postmortem damage by carnivores had destroyed most of the age markers. Data from vertebral ring epiphyseal union contributed to a narrower age estimation when used in conjunction with data from the other skeletal age indicators. Case 2 involved the fresh remains of an unknown female. Union of the vertebral ring epiphyses was consistent with an age range of 18 to 22 years, which was generally compatible with the age ranges obtained from the other skeletal sites (16 to 20 years). From this information, a narrower range of 18 to 20 years was then considered the most likely age at death. For each of the two cases, age information from observations of epiphyseal union of the vertebral centra has (1) been consistent with age information gathered from other skeletal age indicators and or (2) helped narrow down otherwise broader age estimation ranges. This method is recommended for use in estimating age at death from teenage and young adult skeletal remains. PMID- 9854838 TI - Genetic variation at the STR loci D12S391 and CSF1PO in four populations from Austria, Italy, Egypt and Yemen. AB - The short tandem repeat systems (STRs) D12S391 and CSF1P0 were amplified by the polymerase chain reaction (PCR) on blood samples from 100 to 158 unrelated Austrians, Italians, Yemenians and Egyptians. The samples were analyzed by both native and denaturing electrophoresis and two primer pairs were tested for the CSF1PO locus. Except for the CSF1PO data on the Egyptians, no deviations from the Hardy-Weinberg equilibrium were detected. For D12S391, no significant differences were found between the two Arab populations and between the two European populations, but the differences between both Arab populations and the Italians were significant. For CSF1PO, differences were only observed between the Yemenians and all three other populations. No evidence of linkage disequilibrium between the two STRs was found. The observation of a D12S391 allele consisting of only 14 repeats was confirmed by sequencing. PMID- 9854839 TI - DNA short tandem repeat profiling of Chinese population in Taiwan determined by using an automated sequencer. AB - We performed a population study on a Taiwan population using a set of nine short tandem repeat (STR) loci and the amelogenin locus. Allele and genotype frequencies of the STR systems D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820 were determined by polymerase chain reaction followed by automated sequencer analysis. A total of 80 alleles could be observed, using all systems, in a population of 500 individuals. No new intermediate fragments were found in these systems. Allele frequencies showed no deviation from the Hardy Weinberg equilibrium. The mean exclusion power (MEP) ranged from 0.294 for TPOX to 0.711 for FGA (the combined MEP was > 0.999), the discriminating power (DP) ranged from 0.782 for TPOX to 0.964 for FGA, (the combined DP was > 0.99999999949). PMID- 9854840 TI - Italian population data on thirteen short tandem repeat loci: HUMTH01, D21S11, D18S51, HUMVWFA31, HUMFIBRA, D8S1179, HUMTPOX, HUMCSF1PO, D16S539, D7S820, D13S317, D5S818, D3S1358. AB - A population study on thirteen short tandem repeat (STR) loci was performed on 223 unrelated Italian Caucasians. The DNA was amplified by PCR. Separation and detection of the amplified STR fragments was carried out by use of 377 automated system (Applied Biosystems Division/Perkin Elmer). All loci meet Hardy-Weinberg expectations, and the data show only five departures out of seventy-eight pairwise locus tests which is close to expectations of 5% (5/78 = 6.4%). When correcting for multiple tests, there is little evidence for departures from expectations between loci. The combined Power of Exclusion for the thirteen STR loci is 0.99999270. The results demonstrate that these loci will be very useful for human identification in forensic cases in Italy. PMID- 9854841 TI - Tandem repeat structure of the duplicated Y-chromosomal STR locus DYS385 and frequency studies in the German and three Asian populations. AB - The Y-chromosomal short tandem repeat (STR) locus DYS385 can be typed using PCR amplification and separation of the resulting polymorphic fragments by non denaturing high resolution polyacrylamide gel electrophoresis followed by silver staining. The PCR primers amplify a duplicated repeat sequence on the Y chromosome revealing a two-band pattern in male individuals. To determine the internal repeat structure as a basis for a consensus nomenclature, DNA sequence analysis was carried out after subcloning of PCR-amplified fragments revealing the uniform 4-bp repeat structure 'GAAA'. The shortest allele observed consisted of 10 repeat units thus providing the basis for the designation 'allele 10'. Except for isolated point mutations, no systematic differences could be observed either in the repeat sequence or in the flanking regions between the two fragments of a given individual. Thus it was not possible to discriminate between the two loci of the DYS385 system. Four population samples of German (n = 146), Chinese (n = 100), Japanese (n = 100), and Thai (n = 95) origin were studied. In the four groups, alleles 10 to 24 could be observed and genotype frequencies differed significantly. In Germans only one common genotype was present (11-14; 33.8% frequency). In the Asian populations, the frequencies were more evenly distributed with the 13-13 genotype (9%) in Chinese, the 13-17 genotype (14%) in Japanese and the 14-18 genotype (7%) in Thai being the most common. Overall, 69 different genotypes were found, of these 36 were observed in Germans, 36 in Chinese, 33 in Japanese and 44 in Thai. No mutations were detected in 62 father son pairs. Thus DYS385 is a highly polymorphic STR system with population specific genotype distributions. PMID- 9854842 TI - Mechanism from isotope effects. AB - Isotope effects represent perhaps one of the most versatile tools available to investigators interested in the determination of reaction mechanism, particularly in the case of the mechanistic enzymologist. Interpretation of isotope effect data is somewhat more difficult for enzyme reactions, since the chemical or isotope-dependent step(s) is(are) normally not solely rate-limiting as they are for non-enzyme-catalyzed reactions. One can, however, take advantage of rate limitation by multiple steps in an enzyme-catalyzed reaction to obtain information on a number of aspects of mechanism. In this paper, simple theory for the application of isotope effects to reaction mechanism is developed, and applied to organic reactions and those catalyzed by enzymes. Techniques used to measure isotope effects depend somewhat on the isotope used, that is radioisotope vs. stable isotope, or hydrogen isotope vs. heavier atoms. Techniques to be discussed include competitive and noncompetitive (or internal discrimination) measurements. In enzyme-catalyzed reactions, information can be obtained on the order of addition of reactants and relase of products, and this will be illustrated using the 6-phosphogluconate and alcohol dehydrogenase reactions. The use of multiple isotope effects can be used to distinguish between stepwise and concerted reactions, and this will be illustrated with the formate and glucose 6 phosphate dehydrogenase and malic enzyme reactions. PMID- 9854843 TI - Determination of 13C/12C-ratios in rumen produced methane and CO2 of cows, sheep and camels. AB - Naturally produced methane shows different delta 13C-values with respect to its origin, e.g., geological or biological. Methane-production of ruminants is considered to be the dominant source from the animal kingdom. Isotopic values of rumen methane--given in literature--range between -80/1000 and -50/1000 and are related to feed composition and also sampling techniques. Keeping cows, camels and sheep under identical feed conditions and sampling rumen gases via implanted fistuale we compared delta PDB 13C-values of methane and CO2 between the species. Referring to mean values obtained from 4 or 5 samples at different times of 11 animals (n = 47) we calculated delta PDB 13C-medians resulting in small but not significant differences within and significant differences between the species for CO2 and methane. The delta PDB 13C-differences between methane and CO2 were statistically equal within and also between the species. Therefore a linear regression of methane values on CO2 is appropriate and leads to: delta PDB 13C(methane)/1000 = 1.57 * delta PDB 13C(CO2)/1000 - 47/1000 with a correlation coefficient of r = 0.87. PMID- 9854844 TI - Wild rabbit host and some parasites show trophic-level relationships for delta 13C and delta 15N: a first report. AB - We report the first isotopic study of an animal host-parasite system. Parasitic, intestinal nematodes, Graphidium strigosum and Passalurus ambiguus, were 15N enriched relative to their host, the European rabbit Oryctolagus cuniculus, while parasitic cestodes, Cittataenia denticulata and Mosgovoyia pectinata, were 15N depleted, suggesting different trophic relationships. Host embryos were more similar in their delta 13C and delta 15N values to maternal muscle than were any of the parasites. Coprophagy, the direct recycling of food by the rabbit eating its own faeces, did not lead to isotopic differences between stomach contents and faeces, suggesting that the major point for isotopic discrimination in lagomorph nitrogen metabolism is in the animal rather than in the gut. We conclude that bulk delta 13C and delta 15N can reveal valuable new information about host parasite relationships, and these could be explored further at the biochemical level using compound-specific isotopic analyses. PMID- 9854845 TI - Estimation of urea production rate with [15N2]urea and [13C]urea to measure catabolic rates in diabetes mellitus. AB - For verifying catabolic states in insulin-dependent patients and dogs the method estimating urea production rates with 13C and with doubly 15N labeled urea, respectively, has been established. For a fast steady state of urea tracer dilution, a prime of 600 times the continuous infusion rate had to be injected. Urea was isolated from plasma samples by protein precipitation and cation exchange chromatography with a consecutive derivatization of the dried urea fraction (trimethylsilyl derivatives). The masses of the fragment ions m/z 189 (14N14N), 190 (14N15N) and 191 (15N15N) urea are monitored to estimate the [15N2] urea frequency in the overall body urea pool in mol percent excess (MPE). 1 to 15 ng of derivatized urea were measured efficiently. An excellent correlation between expected standard and measured MPE (r = 0.9977) was achieved from solutions containing 1 to 7% [15N2]urea. The interassay coefficient of variation amounted to < 10% for a [15N2]urea portion of > or = 3%. Normoglycemic diabetic patients who were treated with insulin overnight showed significantly higher urea production compared to healthy controls (9.22 +/- 2.07 vs. 5.4 +/- 0.32 mumol.kg 1.min-1; p < 0.05). Measurements in chronic diabetic dogs proved an increased rate of amino acid catabolism (+20% urea production) in systemic versus portal application of insulin in paired studies. This increased nitrogen load in diabetics may accelerate progression of diabetic nephropathy. Thus, the established stable isotope technique may serve as a sensitive and useful indicator of amino acid catabolism in clinical and experimental research. PMID- 9854846 TI - Labelling studies for structure elucidation of a new hydroxymetabolite of tramadol. AB - Tramadol, racemic 1-(3-methoxyphenyl)-2-(dimethylaminomethyl)cyclohexane-1-ol, is an effective analgesic drug. Metabolites of tramadol described so far originate from O- and N-demethylation and are excreted in urine directly or after conjugation. A further metabolite was found in human liver microsome incubations and in the urine of volunteers after ingestion of tramadol. To elucidate the structure of the new metabolite, seven deuterated isotopomers of tramadol have been synthesized and ingested by volunteers. The mass spectra of the metabolites derived showed (i) that it was a hydroxy metabolite, (ii) that the hydroxy group was not located on the aromatic ring, the side chain, or the positions 2 and 6 of the cyclohexane ring, (iii) that the hydroxy-group was introduced to one of the the positions 3, 4 or 5 of the cyclohexane ring. The hydroxy metabolite was formed preferentially from the (-)-enantiomer, (1S,2S)-tramadol. PMID- 9854847 TI - Whole body protein metabolism estimated by 15N tracer experiments and body composition of mice selected for different growth parameters. AB - Whole body protein synthesis was investigated in growing male mice which were long-time selected for high carcass protein amount (DU-6P, protein line) or for high body weight (DU-6, growth line) and in the unselected randomly bred control (DU-Ks). Six mice/line were housed singly in metabolic cages for the estimation of N balance, whole body protein synthesis (end-product method, single dose of 15N-labelled amino-acid mixture), and N distribution in the body. Another six mice/line were used for the determination of the body composition. All mice had free access to a commercial stock diet (crude protein 268 g, gross energy 19 MJ/kg dry matter) and to water. Body weight of both selection lines was about twice that of control mice at the same age. Selection for high body weight resulted in higher body fat content. Scaled to the corresponding body protein pools, the protein synthesis rates of selected mice were significantly higher than in controls, but were not significantly different between both selection lines in contrast to the protein deposition rates. The higher protein accretion in the protein line in comparison to the growth line seems to be due to a combination of a lower protein breakdown and an increased protein synthesis rate. PMID- 9854848 TI - Significance of diagnostic parameters in [13C]octanoic acid gastric emptying breath tests. AB - Two novel characteristic parameters, the latency time (t(lat)) and the ascension time (t(asc)), are proposed for evaluation of non-invasive [13C]octanoic acid breath tests for assessment of the gastric emptying of solids. In breath tests performed in control subjects (n = 30) and diabetic patients (n = 100), the usefulness of these parameters was compared to conventional parameters, i.e., gastric half emptying-time (t1/2,b) and lag phase (t(lag),b). The proposed parameters were only loosely correlated (controls, r = 0.199; diabetics, 0.616). A strong correlation was found between the conventional parameters (controls, r = 0.891; diabetics, r = 0.962). Based on the conventional method, 36 patients were suspicious of delayed gastric emptying including 24 patients which exhibited a simultaneous delay in both parameters. Using the new parameters, a total of 46 patients were suspicious of delayed gastric emptying with 15 and 20 having isolated delay in t(lat) and t(asc), respectively. We conclude that the novel parameters may be more appropriate for examination of the different phases of gastric emptying and for evaluation of gastric emptying disturbances in diabetic patients than the parameters conventionally used for this purpose. PMID- 9854849 TI - Determination of 13CO2/12CO2 ratio by IRMS and NDIRS. AB - Breath tests using 13C-labelled substrates require the measurement of 13CO2/12CO2 ratio in breath gas samples. Next to isotope ratio mass spectrometry (IRMS), which is very sensitive but also complex and expensive, alternatively isotope selective nondispersive infrared spectrometry (NDIRS) can be used to determine the 13CO2/12CO2 ratio in expired breath. In this study we compared NDIRS- with IRMS-results to investigate whether the less expensive and very simply to operate NDIRS works as reliable as IRMS. For this purpose we applicated 1- 13C Phenylalanine to patients with advanced liver cirrhosis and healthy volunteers and took duplicated breath samples for IRMS and NDIRS at selected time points. Our data show a good correlation between these two methods for a small number of samples as required for simple breath tests. Longer series, where repeated measurements are required on the NDIRS instrument lead to a decreasing correlation. This indicates the superiority of IRMS concerning 13CO2-kinetics over longer time periods. PMID- 9854850 TI - Fluorescence imaging and spectroscopy of biomaterials in air and liquid by scanning near-field optical/atomic force microscopy. AB - We have developed scanning near-field optical/atomic force microscopy (SNOM/AFM). The SNOM/AFM uses a bent optical fiber simultaneously as a dynamic force AFM cantilever and a SNOM probe. Resonant frequency of the optical fiber cantilever is 15-40 kHz. Optical resolution of the SNOM/AFM images shows less than 50 nm. The SNOM/AFM system contains photon counting system and polychrometer/intensified coupled charge devise (ICCD) system to observe fluorescence image and spectrograph of micro areas, respectively. Cultured cells were stained with fluorescein isothiocyanate (FITC)-labeled anti-keratin antibody or FITC-labeled phalloidin after treatment with Triton X-100. Fluorescence and topographic images were obtained in air and water. The fluorescence images showed clear images of keratin and actin filaments. The SNOM/AFM is perfect to observe biomaterials in liquid with a liquid chamber while the topographic Images showed subcellular structures which correspond to keratin and actin filaments. PMID- 9854851 TI - The chromatin structure of well-spread demembranated human sperm nuclei revealed by atomic force microscopy. AB - The fundamental structure formed when genomic DNA is packaged by protamine in the human sperm nucleus still remains essentially unresolved. It is known that the binding of protamine, a small arginine-rich protein, to DNA generates a large dense, hydrophobic complex making the sperm chromatin structure difficult to study microscopically. To visualize the internal nuclear structures, isolated human sperm nuclei were swollen extensively in saline buffer using only a reducing agent. The nuclei were swollen during deposition onto coverglass and then imaged in the atomic force microscope (AFM). The two main results obtained from imaging individual well-spread nuclei indicate that native human sperm chromatin is: (1) particulate, consisting primarily of large nodular structures averaging 98 nm in diameter, and (2) also composed of smaller, nucleosome-like particles observed to form linear chains near the nuclear periphery. These two types of chromatin particles imaged by AFM are remarkably similar to other AFM measurements made on native and reconstituted sperm and somatic chromatin. PMID- 9854852 TI - Paleomicrobiological study in dental calculus: Streptococcus mutans. AB - Morphological types of bacterial remains preserved in ancient tartar of teeth from extinct human groups, which included some communities of coastal gatherers, fishermen, hunters, and farmers, and those practicing a mixed economy, were analyzed. Previous studies have shown the presence of bacteria in ancient tartar. The aim of this work was to determine whether Streptococcus mutans was present in ancient populations (500-12,000 years old). Teeth samples were from ancient skulls obtained from different anthropological collections: the north and south of Chile (before the Spanish conquest), Palencia, Spain, and an eastern Mediterranean region (Levant). Optical microscopy showed Gram positive and Gram negative bacteria. Scanning electron microscopy identified morphological types of bacteria. Transmission electron microscopy enabled categorization of bacterial structures. Fluorescence microscopy helped label and identify S. mutans, using polyclonal antibodies. Bacterial morphotypes were related to different subsistence patterns. Hunters, fishermen, and gatherers had a less diverse flora with bacillary and coccal morphotypes. Agricultural groups showed greater diversity with additional filamentous and spiral morphotypes. The best preserved ultrastructural feature was the cell wall. The existence and colonization capacity of the mutans-like streptococci preserved in tartar was established for the ancient populations studied, with the exception of Cerro Sotta (south of Chile). Hence, their occurrence could not be related to diet or subsistence pattern. PMID- 9854853 TI - A micro-Raman spectroscopic study of hydrazine-treated human dental calculus. AB - Hydrazine has been used to remove organic components and to isolate the mineral(s) from human calculus. Micro-Raman measurements were performed on the mineral phase. After the hydrazine-treatment, not only a large reduction in fluorescence but also an increase in Raman signal was observed. The treatment was essential in minimizing thermally-induced chemical changes which could otherwise occur to the original calculus mineral due to the intense laser light. The Raman spectral features of the mineral were nearly all identical among the Raman spectra obtained at many randomly-selected sites by the micro-Raman microbe with a lateral resolution of approximately 1 micron, and were consistent with those of impure hydroxyapatite containing CO3(2-) and HPO4(2-). The spectra contained typical hydroxyapatite bands including PO4(3-)bands of the v1, v2, v3 and v4 modes and one OH- stretch band. Other minor bands due to the CO3(2-) v1 and v3 modes and bands possibly due to the HPO4(2-) v1, v2 and v4 modes were observable by the technique despite the hydrazine-treatment that could in principle remove the HPO4 and CO3 ions from the mineral. In comparison with pure synthetic hydroxyapatite, the intensity of the OH- stretch band relative to that of the PO4(3-) v1 band was approximately 70% weaker, and the bandwidth of the phosphate v1 band was 200% broader, reflecting various crystal imperfections presumably present in the calculus mineral. PMID- 9854854 TI - X-irradiation-induced changes of the prelysosomal and lysosomal compartments and proteolysis in HT-29 cells. AB - As a consequence of external and internal ionizing radiation, lysosome-like bodies have been observed to increase both in size and number in some cell types. We investigated this process by morphological methods (electron microscopy, cationized ferritin uptake, acid phosphatase histochemistry, morphometry) in cultured HT-29 cells. In parallel with these studies, we measured the rate of protein degradation on the basis of 14C-valine release from prelabeled cellular proteins. We found that at 2 and 4 Gy doses of X-irradiation the volume of the vacuolar (probably lysosomal) compartment increased without detectable changes of acid phosphatase activity. A 2 Gy irradiation dose did not change protein degradation rate. However, 4 Gy caused a significant inhibition of 14C-valine release from prelabeled proteins. Our results indicate, that the radiation induced expansion of the lysosomal compartment is not necessarily accompanied by increased lytic activity of HT-29 cells. PMID- 9854855 TI - Cytoskeletal changes during radiation-induced neoplastic transformation of human prostate epithelial cells. AB - We recently reported tumorigenic transformation of SV40-immortalized neonatal human prostate epithelial cells (267B1) by exposure to fractionated doses of X rays. Altered morphology and anchorage independence were observed following two successive fractions of 2 Gy each (F3-SAC). Additional 2 Gy treatments to these non-tumorigenic cells to a total dose of 30 Gy resulted in radiation-transformed tumorigenic colonies (267B1-SXR). Malignant transformation of parental 267B1 cells was also achieved by consecutive 2 Gy exposures to a total dose of 30 Gy (267B1-XR). This study discusses the cytoskeletal changes in the F3-SAC, 267B1-XR and 267B1-SXR derivatives of these human prostate epithelial cells. Confocal and conventional fluorescence microscopy of filamentous actin showed numerous, well organized, evenly distributed stress fibers in the parental cells prior to irradiation, while the anchorage-independent cells and several tumorigenic derivatives exhibited poor stress fiber organization after radiation exposure. This disorganization of actin microfilaments in the radiation-transformed cells was also accompanied by changes in the expression of selective tropomyosin isoforms as judged by two-dimensional gel electrophoresis. These changes in actin organization and tropomyosin expression appear to be coincidental with morphological transformation and acquisition of tumorigenicity in the 267B1 cells following radiation exposure. PMID- 9854856 TI - Atomic force microscopy investigation of radiation-induced DNA double strand breaks. AB - We have used atomic force microscopy (AFM) to study radiation-induced DNA double strand breaks. Double-stranded plasmid DNA was irradiated with 18-MeV electrons in aqueous buffer, using a medical linear accelerator. Doses of 50, 100, 150, and 200 Gy were delivered to DNA samples, and atomic force microscopy was used to measure the length of each DNA fragment. From these measurements, we obtained the average length of the irradiated DNA for each sample and found a linear-quadratic relationship between the average length and radiation dose. PMID- 9854857 TI - A scanning electron microscopy study of Taxus leaves as related to taxonomy. AB - Scanning electron microscopy, when applied to the surfaces of the needles of Taxus spp. (yew) revealed features that appear useful in the taxonomy of this tree species which yields the important anticancer drug, taxol. For instance, all of the four North American species have 3-5 rows of stomata on one-half of the abaxial leaf surface, whereas all of the others, including those from Europe and Asia, have 7-10 rows of stomata. The appearance of individual or fused papilliform epidermal cells and their arrangement on the leaf surface also is a feature that varies between species. Patterns of wax formation appeared on all species of yew examined but none could characteristically be assigned to a given species. Wax pattern variation was dependent upon age, environment, and probably to some extent, species differences. This study may provide some additional useful and reliable indicators in Taxus taxonomy. PMID- 9854858 TI - Hair bundle morphology on surviving hair cells of the chick basilar papilla exposed to intense sound. AB - Exposure to intense sound produces a well-defined "patch" lesion on the chick basilar papilla in which 30-35% of the short hair cells are lost. The present study compares various aspects of sensory hair bundle morphology on surviving hair cells in the patch lesion with hair bundles from matched locations on nonexposed control papilla immediately after removal from the exposure and 12 days post exposure. The height and thickness of the hairs, the total number of hairs in the bundle, the width of the bundle, and the area and perimeter of the apical surface of the hair cell were quantified from scanning electron microscope photomicrographs. An attempt was also made to determine if there was a consistent microstructure to the pattern of hair cell loss within the lesion area. Similar observations in 12-day recovered ears are also presented. The results indicated that stereocilia height increased and width decreased on surviving hair cells in the exposed ear. The width of the hair bundle, the hair cell surface area, and perimeter also decreased. However, the number of hairs per cell remained unchanged, and there was no evidence of any consistent organization to the hair cell loss within the patch across a number of specimens. These observations indicated that the hair bundles on short hair cells underwent changes as a consequence of intense sound exposure. The results after 12 days of recovery were complicated by developmental changes on the papilla and incomplete maturation of the newly regenerated hair cells. It remains to be seen whether these changes were the result of cell sampling in the sound-damaged ear or were due to true structural alterations within the sensory hairs themselves. PMID- 9854859 TI - Scanning electron microscopy of the accessory sex glands of the adult male rat. AB - This study describes the morphology of the accessory sex glands of the adult male rat as observed by scanning electron microscopy (SEM). The purpose was to obtain a systematic and comparative SEM description of these glands and to evaluate different preparation techniques. A common morphological feature is polyhedral delineation of the cells, which exhibited a variable convexity of their apical surface. The cell apices were more or less studded with microvilli. Nevertheless, it was possible to distinguish the glands by their surface morphology. In the ventral prostate, there was a considerable heterogeneity in cell surface appearance. The lateral lobe had a characteristic brush border, and in the dorsal lobe, surface blebbing and intracellular cisternae were observed. The cells of the seminal vesicle were covered by long microvilli, while particularly distinct, elevated cell borders and intracellular cisternae were typical for the coagulating gland. The secretory mechanism was merocrine in the ventral and lateral prostate and the seminal vesicle, and was mainly apocrine in the dorsal prostate. Surprisingly, only merocrine secretion was obvious in the coagulating gland. The most controversial observation, which needs further investigation, was the discovery of large orifices in the apical surface of individual seminal vesicle cells. These orifices may be indicative of an additional apocrine secretion in this gland. In studying this organ system, SEM provides information that adds to previous transmission electron microscopical investigations. PMID- 9854860 TI - Effects of castration upon the morphology of the accessory sex organs of the male rat--a scanning electron microscopy study. AB - A systematic, comparative study of the accessory sex glands of the adult male rat after androgen withdrawal was carried out. The changes were investigated by using scanning electron microscopy at different intervals after surgical castration. The main common signs of epithelial cell involution were flattening of the cell surface, reduction of the size and number of microvilli, some blurring of the cell borders, cessation of secretory activity and diminution of the luminal volume of the glands. Overall, confident signs of atrophy were evident after one week, and complete epithelial involution was reached by the third week. The epithelial cell atrophy was accompanied by a relative stromal hyperplasia. The new observation seems to be that the process of stroma consolidation is progressing for a considerable time subsequent to the completion of the epithelial involution. This phenomenon is particularly evident in the dorsal prostate, the seminal vesicle and the coagulating gland. PMID- 9854861 TI - Combined bronchoalveolar-vascular casting of the canine lung. AB - Canine bronchoalveolar and vascular corrosion casts were prepared using unfixed tissue and Dow-Corning Room Temperature Vulcanizing Silastic 734. The casts were observed using stereo light microscopy and scanning electron microscopy. The casts show the relationship between the vasculature and airway and demonstrate intricate microanatomical details. Also, microvasculature filling was enhanced using unfixed tissue as compared to my previously described technique using fixed tissue. Prewashing the microvasculature with cold phosphate buffered saline appeared to facilitate microvascular filling with silicone rubber. The described method is useful for rapidly making durable models for studying the normal respiratory airway and microvasculature. It should be useful in future studies of diseased pulmonary tissue as well as other normal and diseased tissues where microanatomical relationships between microvasculature and adjacent luminal structures are relevant. PMID- 9854862 TI - Preparation of cultured smooth muscle cells from human myometrium for X-ray microanalysis. AB - Methodological aspects of the use of X-ray microanalysis in physiological and pharmacological experiments on cultured myometrial cells were investigated. Cultured human myometrial cells were grown from biopsies after detaching the fibroblasts. Of the cultured cells, 95-98% showed desmin-like immunoreactivity. Transmission electron microscopy showed that subcultured cells were different from myometrial cells in situ. The effects of washing the cells to remove external salt-rich medium were investigated. All solutions removed the external medium, resulting in lower concentrations of Na and Cl. In the cells washed with 0.3 M mannitol, most of the elemental concentrations were significantly lower than in their unwashed counterparts and those washed in the other solutions. In cells washed in either 0.15 M ammonium acetate or distilled water, no significant differences in P and K compared with their unwashed counterparts were found. There were also no significant differences between cells washed in ammonium acetate and in distilled water. In subsequent experiments ammonium acetate was used. Incubation of cells in standard Ringer's solution resulted in an increase in Na and Cl, and a decrease in K, concomitantly with an increase in Ca. Although Ringer's solution per se can elicit changes in diffusible elements in the cells, physiological and pharmacological effects of oxytocin could still be detected in Ringer's solution. However, effects of oxytocin were different when the experiment was done in culture medium, instead of in Ringer's solution. PMID- 9854863 TI - Relative intranuclear magnesium and phosphorus contents in normal and tumor cells of the human thyroid gland as revealed by energy-dispersive X-ray microanalysis. AB - Energy dispersive X-ray microanalysis was performed on altogether 42 surgically removed tissue specimens of 32 patients, which were taken either from intact thyroid parts or various histopathologically verified tumors of the thyroid gland. The tissue specimens were processed with the freeze-fracture-freeze-drying technique and then analyzed in the so-called bulk specimen form. The studies were carried out during the years 1980-81, when intranuclear monovalent ionic composition was studied in detail. From the retained total elemental peak list, it was possible to calculate retrospectively the relative intranuclear Mg and P contents. The data processed by nested (hierarchical) analysis of variance show that the intranuclear Mg content of the 5 diagnostic groups (normal thyroid tissue, thyroiditis, benign adenomas, differentiated carcinomas and undifferentiated thyroid tumors) increases significantly, in parallel with the increasing malignancy, but the P content remains unchanged. One can conclude that the elevated intranuclear Mg content in the tumors of high malignancy may be of diagnostic importance, and a warning signal for the therapeutic approaches based on Mg-supplementations. PMID- 9854864 TI - Form and function of mammalian lung: analysis by scientific computing. AB - This study approaches the investigation of airway morphology of the lung with a new set of imaging and computer graphical methods, including confocal imaging, computer-guided image acquisition, visualization and fractal graphics. The key result is that, in contrast to the belief that the design of the conductive part of lung of smaller mammals can be described with a trumpet model, the findings reported here document a strongly monopodial branching pattern with the functional consequence of a variation of dead space between the trachea and the acini. This non-dichotomic structural design finds its continuation within the respiratory units as the necessary requirement for an optimal space filling and dense packing which cannot be achieved by a dichotomic branching only. Based on a computer model, computational physics tightly coupled with computer visualistics enables functional simulation of the lung model regarding gas transport. The predicted variance in the ventilation of acini gives rise to an explanation of the well-known difference between the morphologically predicted and physiologically required diffusion capacity. PMID- 9854865 TI - [Conversions and reinterventions in laparoscopic cholecystectomy]. AB - From the introduction of the laparoscopy in our clinic, more and more of the cholecystectomies, reaching over 50% are done by this technique. Based upon the accumulation of an already important experience, the paper tries to analyze the situations in which, during or after laparoscopic cholecystectomy, intraoperative conversions (deliberate or of necessity) or reinterventions were necessary. We present a global view of the number of these cases and also (an in detail) analysis of the causes the imposed such decisions and of the solutions adopted. The percentages of 5.55 conversions and 1.49 reinterventions seem reasonable and acceptable in comparison with the initial results published by some experience surgeons in the field of laparoscopic surgery. PMID- 9854866 TI - [Laparoscopic cholecystectomy in acute cholecystitis]. AB - AIM: Results evaluation of laparoscopic cholecystectomy in acute cholecystitis. MATERIAL AND METHOD: Between 1994-1997 we performed 65 laparoscopic cholecystectomies for histopathologically proved acute lithiasic cholecystitis. We studied clinic and echographic diagnosis, operative moment, conversion rate, operative time, postoperative morbidity and hospitalization. The cholecystectomy was performed within 72 hours in 18 patients (trial I), 4 to 7 days in 25 patients (trial II) and over 7 days in 22 patients (trial III). RESULTS: Diagnosis of acute lithiasic cholecystitis was always possible by clinical examination and ultrasonography. We performed 8 conversions in patients of trial II (2) and III (6). The mean operative time was 68 min. Postoperative morbidity consisted in 4 bile leakages in the liver bed, 1 subhepatic abscess, 5 right pleural effusions. The mean hospitalization was 4.4 days. CONCLUSIONS: Urgent laparoscopic cholecystectomy is a beneficial act for acute lithiasic cholecystitis. The operative moment is the most important factor of influence on conversion rate, operative time and postoperative morbidity. PMID- 9854867 TI - [The conversion of endourological operations: a failure or healthy "clinical thinking"?]. AB - From a lot of 2567 of endourological "low" (TURP, TURB, UIO) and "high" (PNL, URSR, URSA) interventions there are analyzed 48 cases in which there was necessary the conversion to classical surgery. In endourology, just like in laparoscopic surgery, the conversion is divided into conversion of necessity and optional conversion. A reference is made to the way of solving these conversions. The main conclusion is that the urologist has to apply equally and efficiently both classical and endourological procedures. PMID- 9854868 TI - [Total duodenal diversion in hiatal hernia with pathological duodenoesophageal reflux (preliminary results)]. AB - For some patients, reflux disease means also duodenogastric, as well as gastroesophageal reflux; they may suffer because of duodenoesophageal reflux. For these patients, a simple surgical restoration of the cardial competence may prove to be insufficient; on the contrary, an indirect approach, using vagotomy and duodenal diversion may be a good therapeutical option in selected cases. In our hands, total duodenal diversion has already proven to be a good technical solution for the surgical treatment of the postoperative reflux disease, so we decided to expand its first choice indications to hiatal hernia cases, in the presence of duodenoesophageal reflux. Therefore, our study presents the results of the first 7 cases, operated on since 1995. No case had been gastric operated before, but 4/7 had already suffered a cholecystectomy. Duodenoesophageal reflux is discussed: etiopathogenesis related to surgical approach, using standard antireflux surgery (failures of the hiatal techniques and their causes) or total duodenal diversion (technical aspects, antireflux efficiency, secondary functional effects, pros and cons, therapeutical indications). PMID- 9854869 TI - [Urinary infections with hospital germs in general surgery]. AB - Nosocomial urinary tract infection is the most common type of sepsis in the post surgical patient. This paper presents our experience with 218 nosocomial urinary infections (34.93% of our postoperative infections) which complicated the postoperative course of 1002 (21.75%) urethral catheterized patients out of a total of 5950 (3.6%) operated on and under study individuals. Thus we found that urethral catheterization is the most important risk factor for post-surgery urinary infections. Moreover, in our series the postoperative urinary tract sepsis bacteriology is dominated (> 95% of cases) by aerobic gram-negative bacilli that mainly reside in the bowel and also commonly colonize the perineum. Furthermore, we demonstrated that post-surgical urinary infections did not influence directly death rate but they had a significant bearing on care costs. Finally we consider the prevention of postoperative nosocomial urinary tract sepsis as an essential principle of this condition management. PMID- 9854870 TI - [The use of Plastex-type synthetic mesh in the surgery of abdominal wall defects]. AB - The study analyzes a statistical series of 45 patients suffering from large abdominal wall defects and who were managed by plastic surgery with "Plastex" type synthetic mesh, in the period 1 Jan. 1995-1 Apr. 1998. One of the treatment's basic principles is the preserving of the peritoneal pouch from the hernia sac, which must be fixed on the internal side of the synthetic material that replaces the musculoaponeurotic defect. The authors emphasize the low incidence of complications and the absence of recurrences when they used the "Plastex" type synthetic mesh, very well tolerated by the surrounding tissues. For all these reasons, the authors warmly recommend it for plastic surgery in all incisional hernias, because their coming out shows the poor quality of the local musculoaponeurotic material. PMID- 9854871 TI - [A case of disseminated intravascular coagulation--a paraneoplastic manifestation of a borderline ovarian tumor]. AB - A 24-year-old-woman was admitted because of disseminated intravascular coagulation (DIC), menometrorrhagia and galactorrhea. The investigations performed showed a right adnexal tumor after the equilibration of DIC with plasma substitution, we performed a right adnexectomy with limited excision of peritoneal. The pathologic examination showed a focus of endometriosis on the right ovary who had a polycystic look and a right adnexal fibromyxoma (premalignant lesion). The post operative evolution was good, with the loss of entire onco-hemato-endocrinologic picture. We describe the interrelation between DIC (paraneoplastic syndrome), menometrorrhagia and galactorrhea, the pathologic hypothesis and the treatment of DIC. PMID- 9854872 TI - [Rectal lesions in children. Clinico-therapeutic considerations of 12 cases]. AB - The lesions of the anus and the rectum are not very really found in children, representing even a appreciable percentage; in the Pediatric Surgery Clinic of the University Hospital no. 1 of Craiova were admitted and operated 12 patients with lesions of the rectum between 1986 and 1998. The causes of these lesions were: polytraumatism--3, falls in different sharp things--7, rectal perforations on the septic base from the peritoneal cavity--1, iatrogenic cause--1. The surgical treatment were consisted in: the suture of wounds on the perineal way, lavage, drainage, antibiotherapy--6 cases, the iliac left anus with the re establishing of the continuity--3 cases, the suture of wounds on the perineal and abdominal ways--2 cases. The conditions were good in 9 cases--cured, and 3 deceases (2 polytraumatism, 1 case malformed new-born with perforation produced by using haegar). PMID- 9854873 TI - [Occlusive leiomyomas of the small intestine]. AB - The authors report two leiomyomas of small intestine compressive tumoral mass. Aspects of symptomatology, histopathology, surgical management and prognostic factors with reference in childhood are discussed. PMID- 9854874 TI - [Term pregnancy, a factor favoring the bleeding of a gastroduodenal ulcer. A case report]. AB - We present the case of a 31-years-old woman, in the 37th week of pregnancy who was urgently hospitalized in our section, with superior digestive bleeding, in a serious state, externalized in hematemesis and melena. The pregnancy being in due time, with sings of secondary fetal pain, probably due to mother's serious anemia, we proceeded to the extraction of the foetus using caesarean section, followed by hemostasis "in situ" of a bleeding duodenal ulcer and the excision of another healed ulcerous lesion (ulcer in mirror). The postoperative evolution was favorable both for mother and new-born child. Reexamined after two years, the patient didn't present subjective pains of an ulcerous type any more, neither lesions visible at an endoscopic examination. PMID- 9854875 TI - Cryptosporidium and the safety of our water supplies. PMID- 9854876 TI - Enhanced surveillance of tuberculosis in England and Wales: circling the wagons? PMID- 9854877 TI - Universal or selective immunisation against hepatitis B virus in the United Kingdom? A review of recent cost-effectiveness studies. AB - There is still no consensus on which hepatitis B virus (HBV) immunisation option should be adopted in the United Kingdom (UK). This review considers why three recent UK studies on the subject reached different conclusions, and whether they provide sufficient information to base an informed decision on cost-effectiveness grounds. The studies differed in methodology, particularly in the models used to estimate the effectiveness of the competing programmes. This led the authors to draw very different conclusions as to the relative cost-effectiveness of universal infant and selective immunisation, probably because the study that favoured infant immunisation omitted an allowance for the indirect protection afforded to others by immunisation of a proportion of the population. This would lead to the underestimation of the relative effectiveness of a programme targeted at high-risk individuals. Selective vaccination is probably more cost-effective than mass immunisation, but universal immunisation may still be considered a cost effective option (in addition to selective immunisation) if future health benefits are not discounted (i.e., given a lower value than present ones). If future health benefits are discounted then mass infant immunisation is almost certainly not cost-effective. If selective immunisation is to be adopted, then the current (selective) strategy should be properly implemented, as it appears to have had little impact on HBV infection and disease. PMID- 9854878 TI - Guidelines for optimal surveillance of Clostridium difficile infection in hospitals. AB - The availability of surveillance data on C. difficile infection in hospitals in England and Wales is being jeopardised by the trend not to culture the organism for diagnostic purposes. NHS trust laboratories that no longer have the ability to isolate C. difficile cannot investigate putative outbreaks or monitor antimicrobial susceptibilities. These laboratories may now need to rely on their local public health laboratory for such investigations. Recent recommendations from the Department of Health(DH)/PHLS have highlighted the need for culture in outbreak investigations, for surveillance purposes, and for monitoring antimicrobial susceptibilities. It is important, therefore, that NHS diagnostic laboratories and public health laboratories, in particular, retain the ability to isolate C. difficile. A cost-effective approach is described that will facilitate surveillance by typing of strains and also enable their antimicrobial susceptibilities to be monitored. PMID- 9854879 TI - Molecular characterisation of Cryptosporidium parvum from two large suspected waterborne outbreaks. Outbreak Control Team South and West Devon 1995, Incident Management Team and Further Epidemiological and Microbiological Studies Subgroup North Thames 1997. AB - Polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis of the Cryptosporidium parvum outer wall protein (COWP) gene was applied to specimens collected from 95 patients with cryptosporidiosis associated with two suspected waterborne outbreaks, 46 sporadic human cases and 62 infected livestock from other areas, and 12 patients infected with other gastrointestinal parasites. Ninety-six per cent of C. parvum isolates from patients linked to the two suspected waterborne outbreaks were of genotype 1; all the isolates from livestock were of genotype 2. Isolates from 59% of the sporadic human infections were of genotype 1 and 35% were of genotype 2. Specimens from two patients yielded both genotypes. Specimens from patients infected with other parasites yielded no amplicons. PMID- 9854880 TI - Investigation of an outbreak of cryptosporidiosis associated with treated surface water finds limits to the value of case control studies. AB - Fifty-two cases of cryptosporidiosis satisfied the case definition employed in investigation of an outbreak in Spring 1996 among residents of the Wirral peninsula supplied by a single water treatment plant using river water. The attack rate among those whose water was supplied solely from the plant was 1.42 per 10,000, compared with 0.42/10,000 among those having some but less than 50% supplied from it. Single oocysts were detected in treated water from this plant on four occasions during the investigation. A case control study did not demonstrate a significant association between illness and water consumption and no obvious failure in water treatment procedures occurred during the relevant period. Nevertheless, according to PHLS criteria, this outbreak was strongly associated with water, as the descriptive epidemiology was consistent and oocysts were detected, albeit in small numbers, in treated water. This paper discusses the value of detection of oocysts and case control studies in investigating waterborne outbreaks of cryptosporidiosis. Populations that normally drink surface water, particularly river water, are thought to be exposed repeatedly to cryptosporidium oocysts and so develop some protective immunity. Case control studies are therefore likely to have less power in the investigation of outbreaks in localities where the population normally drink treated surface water. Although oocysts are often isolated from treated surface waters without being associated with obvious disease in the population, their detection should still be considered in assessing the strength of association of waterborne outbreaks. PMID- 9854882 TI - Influenza surveillance in England and Wales: October 1997 to June 1998. AB - Influenza caused low levels of mortality and morbidity in England and Wales in the 1997/98 season. Influenza viruses of the H3N2 and H1N1 subtypes were isolated in small numbers from community and hospital patients in November and December. Their numbers subsequently increased to peak in February, H1N1 a week or two before H3N2. Most of the H1N1 isolates were similar to the A/Bayern/7/95-like virus, included in the vaccine recommended for 1997-98, whereas most H3N2 isolates were A/Sydney/5/97-like, a virus not covered by the season's vaccine but recommended for 1998/99. Consultation rates in general practice for influenza like illnesses remained low until early February, when a small increase coincided with the increase in laboratory confirmed influenza. Fewer deaths directly attributed to influenza were registered during the surveillance period than in recent years. Localised outbreaks occurred in schools and nursing homes towards the end of the winter. PMID- 9854881 TI - A large outbreak of cryptosporidiosis associated with a public water supply from a deep chalk borehole. Outbreak Investigation Team. AB - Three hundred and forty-five confirmed cases were reported in a large waterborne outbreak of cryptosporidiosis in North Thames in the spring of 1997. The descriptive epidemiology, attack rates, a case control study, and the detection of oocysts in the water suggested strongly that the outbreak was associated with drinking unboiled tap water that originated from one deep chalk borehole. The 746,000 people living in the water distribution area were advised to boil their drinking water. Investigations did not reveal how oocysts entered the borehole. This is the first published report of a cryptosporidium outbreak caused by filtered borehole water and we believe it to be the largest outbreak due to groundwater to have been reported. Borehole supplies are regarded as relatively pure sources of water and this outbreak has implications for the future monitoring and treatment of drinking water extracted from boreholes. PMID- 9854883 TI - Legionnaires' disease in residents of England and Wales: 1997. AB - Two hundred and twenty-six residents of England and Wales who developed legionnaires' disease with onset of infection in 1997 were reported to the PHLS Communicable Disease Surveillance Centre. Twenty-eight cases (12%) were reported to have died. Half of the reported cases (114) were associated with travel, either abroad or in the United Kingdom. Six cases acquired infection in hospital and the remaining 106 were presumed to have acquired infection in the community. Nineteen cases were linked to industrial or community associated outbreaks or clusters in England, two cases to a hospital outbreak, and 25 cases to outbreaks or clusters abroad. The remaining 180 cases (80%) were reported as single cases. The proportion of cases diagnosed by detection of urinary antigen has continued to increase. In 1997 this method of diagnosis was included for 60% of confirmed cases and in 46% of all cases. PMID- 9854884 TI - Legionella infections in Scotland: 1997. PMID- 9854885 TI - A cluster of tuberculosis cases in a family. AB - A 40 year old Indian woman who had recently returned from a visit to India with her eldest son was diagnosed with miliary tuberculosis in January 1994. Contact tracing revealed that her three sons, aged 19, 21, and 22, all had BCG scars from vaccinations administered at birth in India and Heaf tests of grades 2-3. None of them was given chemoprophylaxis, although this was considered, as suggested by the guidelines then available (1990). About 18 months later, the eldest son was notified as a case of tuberculosis. The youngest son, who shared a room with the eldest, developed a right tuberculous pleural effusion over three years after his mother became ill. The 41 year old husband of the index case and their 21 year old son remain well at the time of this report and are being followed up. These cases highlight the importance of stringent follow up, contact tracing, good liaison between general practitioners and chest clinic staff, and chemoprophylaxis for the contacts of tuberculosis cases. Guidelines about the management of contacts of sputum positive cases over 16 years old need to be clarified. A history of a recent overseas trip should raise suspicions of infection with tuberculosis. PMID- 9854886 TI - Optimisation of the protocol for detection of Aeromonas species in ready-to-eat salads, and its use to speciate isolates and establish their prevalence. AB - Aeromonas spp. are detected in more than 500 cases of gastrointestinal infection each year in England and Wales. This study aimed to identify their prevalence in ready-to-eat salads, which are a potential source of aeromonas infection. The protocol for isolation of mesophilic Aeromonas spp. from salads was optimised. Using the improved method, Aeromonas spp were isolated from 19 of 25 samples (25 g) of ready-to-eat salad products. Aeromonas organisms were counted, isolates were identified to species level, and the effect of pH on colonisation of salads was assessed. Aeromonas was present at high levels in six salads (> or = 100 cfu/g). The major species present in salads was Aeromonas caviae, but A.hydrophila and A.sobria, which have more pathogenic potential, were also isolated. It is hoped that this study will help to assess the risk to public health of aeromonas in salads. PMID- 9854887 TI - The risk to public health of aeromonas in ready-to-eat salad products. AB - Mesophilic Aeromonas spp. are isolated regularly from cases of gastrointestinal infection and have markers indicative of enteropathogenicity. Is aeromonas, which is present in a large proportion of ready-to-eat salads, actually a gastrointestinal pathogen? Isolates of mesophilic aeromonas from salads were characterised in terms of their ability to grow at refrigeration temperatures over the given shelf life and by the presence of markers of potential virulence. The major phenospecies present in salads, A.caviae, showed little enteropathogenic potential. Thirty-five per cent of aeromonas salad isolates are A.hydrophila or A.sobria, however, and all isolates tested had at least one marker of enteropathogenicity, including cytotoxin and haemolysin production, adherence to epithelial cells, and resistance to certain antibiotics Despite the presence of markers of enteropathogencity, the lack of epidemiological evidence of a link between infectious intestinal disease and the consumption of salads suggests that their contamination with aeromonas does not pose a significant risk to health in immunocompetent adults. PMID- 9854888 TI - Results of a survey of diagnosed HIV infections prevalent in 1996 in England and Wales. AB - This paper describes a survey undertaken to assess the caseload of HIV infected patients who received medical care from statutory service providers in England and Wales in 1996 in order to inform health authorities about the size and composition of their resident population of such patients. A total of 13,670 HIV infected patients were identified as living in England and Wales, 70% of whom lived in the Thames NHS executive regions. Over half the total caseload received care within their health authority of residence. Regional care centres attracted patients from wider areas, however, particularly in the North West and Thames regions. This survey of prevalent diagnosed HIV infections, one of a series conducted annually, provides public health specialists with information relevant to their localities without compromising patient confidentiality. Along with other data from the surveillance of AIDS cases and HIV infections it contributes to the assessment and projection of demands on health and social services and provides evidence on which to develop and direct national and local health campaigns. PMID- 9854889 TI - Using age to predict cervical infection in women presenting with lower genital tract syndrome in Cape Town. AB - In developing countries without access to laboratory testing, the syndromic management of cervicitis in women with lower genital tract symptoms results in overtreatment. We evaluated the validity of using different age cut-offs to predict the risk of cervical infection in women attending a clinic for sexually transmitted diseases in Cape Town. Specimens from 161 women with lower genital tract symptoms were examined microbiologically for evidence of cervicitis (gonorrhoea and/or chlamydial infection). Women with cervicitis (median 21 years) were significantly younger than those not infected (median 23 years). Retrospective simulation using the age cut-offs (< 45 years (current approach), < 30 years, and < 25 years) to predict cervicitis showed that specificity could be increased (overtreatment reduced) from 17% to 50% if presumptive diagnoses of cervicitis were restricted to women aged under 25 years. Lowering the age cut-off would severely limit sensitivity, however, and as many as 9% and 20%, respectively, of infected women would go untreated if the age cut-offs < 30 and < 25 years were used. The use of a lower age cut-off would therefore serve neither individual nor public health in the presumptive diagnosis and treatment of cervicitis in a high risk population, and would result in a substantially lower detection rate. PMID- 9854890 TI - Shigella outbreak in a school associated with eating canteen food and person to person spread. AB - In June 1993 an outbreak of Shigella sonnei infection at a primary school in south east England affected 42% of 327 pupils and staff. Attack rates of diarrhoea and fever were 33% for children aged 4 to 8 years, and 8% for those aged 8 to 12 years (p < 0.00001). Illness was associated with eating canteen food (relative risk 5.9; 95% confidence interval 3.4, -10.3). All strains examined were S. sonnei phage type 3, with the same antibiogram (ttSTSS), and were indistinguishable using colicin typing and biotyping (colicin type 9, E8) and pulse field gel electrophoresis. Molecular epidemiology suggested but could not confirm that the outbreak strain was introduced into the school population from the community. PMID- 9854891 TI - Improving the uptake of selective neonatal BCG immunisation. AB - A survey of selective neonatal BCG immunisation in Bolton in 1993 showed that only 6% of babies in the defined risk groups received BCG. Community medical officers at that time were responsible for administering the BCG. Midwives were trained to take on this role. In 1994, 88% of babies in the defined groups were immunised. This figure has remained stable in subsequent surveys and we are now aiming for 95% uptake. PMID- 9854892 TI - Screening child playgroup contacts of an adult with smear negative tuberculosis. AB - A childcare assistant with smear negative, culture positive tuberculosis worked for four months with 85 children who attended playgroups for 2.5 to 5 hours each week. Eighty-two of the children completed contact tracing procedures. No evidence was found of transmission of disease to any child. PMID- 9854893 TI - A user's guide to producing and interpreting tree diagrams in taxonomy and phylogenetics. Part 4: Practice. PMID- 9854894 TI - Employment of industrial year students--pain or gain? PMID- 9854895 TI - Fire awareness in the office and laboratory. AB - Three elements are needed for a fire to start and flourish: heat, fuel, and oxygen. These elements form the three sides of the 'fire triangle', and the removal or isolation of any one will extinguish a fire. Such action forms the basis of fire fighting techniques currently in use by the fire authorities. Two sides of the fire triangle, fuel and oxygen, are always present in the workplace. This article looks at the common causes of fire in the workplace, and the precautions and control measures needed to prevent it. PMID- 9854896 TI - Sewage contamination of drinking water in Bolton. PMID- 9854897 TI - Waterborne cryptosporidiosis. PMID- 9854898 TI - Cell cycle control protein p34cdc2 homologue and its phase specific accumulation in synchronized onion (Allium cepa L.) root meristem cells. AB - A key regulator of the cell cycle is a highly conserved protein kinase whose catalytic subunit, p34cdc2, is encoded by cdc2 gene. Immunoblotting with a polyclonal antibody raised against PSTAIRE sequence (found in the N-terminal region of all cdc2 and cdc2 related proteins throughout the phylogenetic scale including higher plants), was used to study the presence of p34cdc2 in onion scale leaves and root tip cells. p34cdc2 homologues are beyond the detection level in scale leaves. PSTAIRE antibody was used to estimate p34cdc2 kinase protein levels during cell cycle in highly synchronous population of Allium cepa L. root meristem cells. p34cdc2 kinase protein showed gradual increase in their levels from S phase to G2 phase boundary. Immunoprecipitation followed by in vitro histone H1 kinase assays also depicted that its kinase activity increased parallel to the increase in p34cdc2 level. PMID- 9854899 TI - Modulation of biochemical activity of hepatocytes in culture by matrix substratum. AB - The influence of extra cellular matrix on the biochemical activity of hepatocytes was studied by maintaining rat hepatocytes in primary culture in a serum free medium on different matrix protein substrata or biomatrices prepared from liver, aorta or mammary gland. There was significant difference in the individual protein synthesis and distribution by cells maintained on different substrata. Comparison of the kinetics of synthesis and secretion of albumin by cells maintained on different tissue biomatrix showed that those maintained on hepatic biomatrix synthesized more albumin and retained more of albumin synthetic capacity, when compared to those maintained on aortic and mammary gland biomatrix. Similarly, hepatocytes maintained on hepatic biomatrix synthesized significantly more apo B, the major apo protein of VLDL, than those maintained on heterologous tissue matrix. Induction of tyrosine aminotransferase by dexamethasone and the uptake of [14C]-amino isobutyric acid were found to be maximum in cells maintained on liver biomatrix than the heterologous biomatrix. But cells maintained on hepatic biomatrix incorporated less amounts of radioactivity into total cytoskeletal proteins as well as the individual proteins such as actin and the cytokeratins C8 and C18 while that by cells maintained on aortic biomatrix was significantly high. Quantitative analysis of the relative incorporation of radioactivity into individual cytoskeletal proteins and albumin in pulse labelling studies with cells maintained in culture on different matrix for different lengths of time revealed a reciprocal relationship between these two activities. These results indicate that the substrata with which the cells are in contact influence on a selective basis, the biochemical activity of hepatocytes in primary culture. PMID- 9854900 TI - Copper(II) as an efficient scavenger of singlet molecular oxygen. AB - Reactive oxygen species (ROS) are considered to play an important role in tissue injury that damages DNA, proteins, carbohydrates and lipids. Increased production of ROS and/or decreased efficiency of antioxidant defense system has been shown to contribute to a number of degenerative processes including cancer and AIDS. Among the various forms of ROS, singlet oxygen (1O2), which is generated predominantly in photosensitization reactions, is of particular physiologic significance because of its selectively long life in aqueous solution, its ability to cross the cell membrane barrier and high reactivity towards biomolecules. In the present study, the 1O2 scavenging potential of Cu(II) has been evaluated by (i) generating 1O2 by photosensitization of rose bengal (RB), (ii) establishing 1O2 quenching with recognized 1O2 scavengers like sodium azide, DABCO and (iii) examining the effect of Cu(II) in scavenging of 1O2. The results revealed that Cu(II) inhibited the rate of 1O2 production by 88%, 68%, 40%, 21% and 10% at a concentration of 10(-2) M, 5 x 10(-3) M, 10(-3) M; 5 x 10(-4) M, and 10(-4) M, respectively. Under similar experimental condition, sodium azide or DABCO at 10(-2) M inhibited the 1O2 production by 86% and 88%, respectively. Other 1O2 generating photosensitizer like hematoporphyrin, riboflavin and methylene blue also produced identical results with Cu(II) but Fe(II), Fe(III), Zn(II) or As(III) did not produce any quenching of 1O2. Presence of a copper binding peptide (Gly-Gly-His) in the reaction system reduced the 1O2 scavenging capacity of Cu(II) by 52-66% depending upon the UV dose. The 1O2 scavenging property of metal ion appears to have an advantage to reduce the oxidative damage of photodynamic reactions in order to prevent ROS-induced toxicity reactions. PMID- 9854901 TI - Modulation of bilayer fluidity by lipid peroxidation of human placental syncytiotrophoblast membranes during embryogenesis. AB - Placental syncytiotrophoblast while regulating the passage of nutrients from maternal blood to the fetal circulation exposes itself to the risk of oxidative attack by the oxygen free radicals. Extent of lipid peroxidation (LPO) investigated in placental brush border membrane (BBM) and basal membrane (BM) revealed a decreasing trend with gestational progress. Steady-state fluorescence anisotropy measurement of 1,6-diphenyl-1,3,5-hexatriene (DPH)-labeled malondialdehyde treated placental membrane vesicles suggested modulation of BBM and BM fluidity by lipid peroxidation in all gestational ages. alpha-tocopherol content in both the placental membranes which increased as gestation progressed has been proposed to play a significant role in decreasing LPO of placental membranes during intrauterine development. PMID- 9854902 TI - Hormonal regulation of malate-aspartate shuttle enzymes during postnatal development of mice. AB - The regulation of malate-aspartate shuttle enzymes by hydrocortisone was studied in the liver and kidney of 15-, 30- and 60-day old mice. It has been observed that adrenalectomy decreases and hydrocortisone treatment to adrenalectomized mice increases the activity of cytosolic and mitochondrial malate dehydrogenase and cytosolic aspartate aminotransferase in the liver of 15-, 30- and 60-day old mice. Per cent decrease following adrenalectomy remained almost similar in all the postnatal ages studied. However, hydrocortisone effect shows age-dependency. In the kidney, adrenalectomy decreases and hydrocortisone treatment increases the activity of these isoenzymes only in 30- and 60-day old mice. These findings entail that the same enzyme in different tissues of developing animals may be regulated differentially by the same physiological stimuli. Our findings on hormonal regulation of malate-aspartate shuttle enzymes show that they are subjected to different physiological control in various tissues during postnatal development of mice. PMID- 9854903 TI - Reversible inhibition by pyridoxal 5'-phosphate and irreversible inactivation by urea and guanidine hydrochloride of thymidylate synthase from Lactobacillus leichmannii. AB - Pyridoxal 5'-phosphate reversibly inhibited thymidylate synthase from Lactobacillus leichmannii. The inhibition was competitive with dUMP (Ki = 1 microM) and non-competitive with 5,10-CH2-THF (Ki = 0.08 microM). Treatment of native or pCMB-treated enzyme with urea (5 M) or guanidine hydrochloride (4 M) resulted in inactivation and dissociation of the homodimer (74 kDa) into monomer (37 kDa). PMID- 9854904 TI - Synthesis of NAD+ in erythrocytes incubated with nicotinic acid and the effect of di-(2-ethyl hexyl) phthalate (DEHP). AB - Synthesis of NAD+ from nicotinic acid by erythrocytes incubated in SAGM phosphate solution and effect of di-[2-ethyl hexyl] phthalate, a plasticizer commonly used in PVC blood/component storage bags, on this synthesis was studied. Erythrocytes are able to synthesise NAD+ in SAGM (sodium chloride, adenine, glucose, mannitol) phosphate solution and this synthesis was more in the presence of added nicotinic acid (optimum concentration 1 mM). The level of NAD+ decreased when the incubation period was increased from 24 to 48 hr. Glutamine had a deleterious effect on this synthesis, possibly due to the decrease in pH. Di-[2-ethyl hexyl] phthalate had an inhibitory effect on NAD+ synthesis when the cells were incubated in SAGM phosphate solution, either alone or in the presence of added nicotinic acid. There was significant decrease in the release of potassium and haemoglobin from the cells in the presence of nicotinic acid, indicating increased red cell stability. PMID- 9854905 TI - Spectrophotometric and spectrofluorometric studies on interaction of cationic dyes with bacterial capsular polysaccharide. AB - Interaction of Klebsiella K14 capsular polysaccharide with cationic dyes pinacyanol chloride, acridine orange and phenosafranin has been studied by spectrophotometric and spectrofluorometric techniques. The polymer containing both glucuronic acid and pyruvic acid in its repeating unit behaved as a unique polyelectrolyte. It induced blue shift of the absorption band of pinacyanol chloride indicating strong metachromasy. Stoichiometry of the polyanion and the dye cations in the polymer-dye compound (1:2) indicated that both glucuronic acid and pyruvic acid acted as potential anionic sites for interaction with the cationic dye molecules. The stoichiometry of anionic site (of polyanion): cationic site (of dye) in the polymer dye compound was calculated as 1:1. Interaction of the polymer with acridine orange and phenosafranin dyes studied by fluorescence measurements demonstrated Stern-Volmer type of quenching. Equivalent weight of the polymer was determined by spectrophotometric and spectrofluorometric titrations. From the present studies chromotropic property of the polymer was established. PMID- 9854906 TI - Safranine-O as membrane potential probe: a mechanistic study using fluorescence spectroscopy. AB - Use of safranine-o has been examined as membrane potential probe in 1-palmitoyl-2 oleoyl-3-phosphatidylcholine (POPC) vesicles both in presence and absence of cholesterol. The fluorescence signal increases in presence of vesicles and the increase in fluorescence intensity on hyperpolarization with valinomycin is diffusion potential dependent. The fluorescence spectra recorded after time driven experiments reveals the blue shift in gamma max of fluorescence with increasing diffusion potential. The fluorescence spectra of vesicles-associated dye is at variance with those of the safranine-o in organic solvents. In organic solvents with increasing hydrophobic character of the solvent the gamma max is slightly red shifted. The electronic spectra of the dye molecule and the charges on different atomic centers have been calculated by quantum chemical method GRINDOL. The predicted first excited state originating from the phenazine moiety is in very good agreement with the excitation wavelength. On the basis of charges on various atoms the binding of safranine with vesicles has been discussed. The nonlinear behaviour of fluorescence signal with delta phi, anisotropy measurements and the computational results, reveal the penetration of bound dye molecules (along with orientation) as a function of diffusion potential. Addition of microaliquots of 1.5 M K2SO4 to already hyperpolarized vesicles decreases the fluorescence signal and the fluorescence spectra recorded on stabilization of signal after each addition showed a shift in gamma max of fluorescence in opposite direction i.e. red shifted. PMID- 9854907 TI - Healthier, wealthier and wiser. PMID- 9854908 TI - Specialties and the code. PMID- 9854909 TI - Curbing implant complications. PMID- 9854910 TI - Sensitivity and Class II restorations. PMID- 9854911 TI - Simplifying extractions. PMID- 9854912 TI - 'Quick-fix' technique. PMID- 9854913 TI - Mice provide clues to osteoporosis. PMID- 9854914 TI - Painkillers to carry warning labels. PMID- 9854916 TI - Are you online? PMID- 9854915 TI - Watching what your patients eat. PMID- 9854918 TI - The economic state of dentistry. Demand-side trends. PMID- 9854919 TI - Dentists and their practices. PMID- 9854920 TI - Dentist work force and educational pipeline. PMID- 9854921 TI - Women's oral health awareness and care-seeking characteristics: a pilot study. AB - The authors designed a pilot study to examine issues that affect women's oral health. They found a substantial lack of awareness regarding important oral health issues, and that 44 percent of the participants reportedly did not have regular dental care. High perceived dental need was reported concurrently with low dental care attendance. Additionally, reported lack of dental insurance and self-perception of low income and of poor dental health were important explanatory factors for the women who reportedly did not have regular dental care. PMID- 9854922 TI - Inhibiting interspecies coaggregation of plaque bacteria with a cranberry juice constituent [published erratam appear in J Am Dent Assoc 1999 Jan;130(1):36 and 1999 Mar;130(3):332]. AB - Dental plaque stability depends on bacterial adhesion to acquired pellicle, and on interspecies adhesion (or coaggregation). A high-molecular-weight cranberry constituent at 0.6 to 2.5 milligrams per milliliter reversed the coaggregation of 49 (58 percent) of 84 coaggregating bacterial pairs tested. It acted preferentially on pairs in which one or both members are gram-negative anaerobes frequently involved in periodontal diseases. Thus, the anticoaggregating cranberry constituent has the potential for altering the subgingival microbiota, resulting in conservative control of gingival and periodontal diseases. However, the high dextrose and fructose content of the commercially available cranberry juice makes it unsuitable for oral hygiene use, and the beneficial effect of the high-molecular-weight constituent requires animal and clinical studies. PMID- 9854923 TI - The influence of dental unit design on percutaneous injury. AB - The handpiece receptacle of a European, buggy-whip-style dental unit is in a different location than that of a conventional dental unit. This study investigated whether this difference affects the incidence of percutaneous injuries among dental professionals. The researchers asked dental professionals to record descriptions of percutaneous injuries they sustained during a period of 30 workdays. Findings indicated that most injuries were bur-related and that there was no statistically significant difference between the European and the conventional dental units with respect to the incidence of percutaneous injury. PMID- 9854924 TI - Affordable implant prosthetics using a screwless implant system. AB - Many dentists have been reluctant to place dental implants because they have found that most implants are costly and time-consuming to place and have long term maintenance problems. Most of these problems are caused by using screws to connect the abutment to the implant, the crown to the abutment or both. The use of a screwless implant system and conventional prosthetics, the author contends, can make implant dentistry affordable, versatile and easy to incorporate into all general dental practices. PMID- 9854925 TI - Whiplash and temporomandibular disorders: a critical review. AB - Some authors have hypothesized a relationship between rear-impact motor vehicle collisions and subsequent symptoms of neck pain and temporomandibular disorders, or TMD, despite no facial impact. This article examines the TMD aspect in terms of the physiological basis and cultural factors influencing the reporting of such symptoms. PMID- 9854926 TI - Providing endodontic care for teeth with ceramic crowns. PMID- 9854927 TI - A profile of current Internet users in dentistry. AB - The authors developed a profile of current Internet users in dentistry using a survey administered through electronic mail and the World Wide Web. Eight hundred twenty-five respondents from 52 countries comprised dentists, assistants, hygienists, dental students and educators. Respondents reported that they used the Internet for discussing clinical cases, obtaining diagnostic and therapeutic information, buying dental products, communicating with patients and participating in continuing education. Eighty percent considered the Internet to be a useful or very useful resource in dentistry. PMID- 9854928 TI - ERISA update. Employee Retirement Income Security Act. PMID- 9854929 TI - Amalgam vs. composite resin: 1998. AB - Class II resin restorations have been evolving in American dentistry for 30 years, but the concept has had significant difficulty being accepted because of stigma attached to early generations of composites. Currently available composite resins for posterior tooth restorations have physical characteristics justifying their use. Techniques for Class II resin placement have improved significantly, and mastery of them is within the ability of both dentists and dental students. Although composite resin materials and techniques present clinical challenges, so do amalgam materials and techniques. It is time to accept Class II resin restorations, improve dentist and student education about their use, increase acceptance by third-party organizations and various approving groups, and bring this concept into the mainstream of U.S. dentistry. PMID- 9854930 TI - Facing the new century with the National Institute of Dental and Craniofacial Research. PMID- 9854931 TI - [Extensive right-sided hepatectomy for a metastatic process in patients operated on for colonic carcinoma]. AB - Proceeding from experience with a large number of liver resections performed for single or multiple metastases atypical in appearance, including left lobectomies, a description is presented of wide right-sided hemihepatectomy, done for the first time (13 May 1997) in the surgical clinic of the National Oncological Center. The intervention is undertaken because of two metastases in the liver subsequent to radical operation for carcinoma of the sigmoid colon. The single stages of the operation are likewise described. The effectiveness of application of Penchev's "Parenchymatome" (made in Bulgaria--Military Medical Academy) and "Tisomat" apparatus is evaluated. Using the latter the resection liver surface is sprayed with a bicomponent fibrin adhesive "Tisucol", obtained from the firm Immuno-AG. PMID- 9854932 TI - [The surgical treatment of cervical lymph node metastases in differentiated carcinoma of the thyroid (DCTG)]. PMID- 9854933 TI - [Local recurrences and mortality after mastectomy--a retrospective analysis]. AB - It is the aim of the study to reveal the underlying causes of local recurrences and lethality in post-mastectomy patients. A retrospective analysis is done in a series of 293 patients presenting mammary gland carcinoma, operated in the period October 1994 through June 1996. The causes of local recurrences and lethality are linked to the stage of disease (TNM-system), pathohistological patterns of the neoplasm, malignancy degree, as well as inadequate surgical and therapeutic approach against the background of concomitant affections and age factor. The analysis is done using data-base elaborated by the author in computerized program "Paradox 5 for Windows". As shown by the results, in patients with high-degree neoplastic malignancy, inaccurate assessment by the TNM system preoperatively and in 2b and following stages, the rate of local relapses and lethality augment. 1. A more accurate elucidation of the preoperative stage of the disease using instrumental methods is absolutely necessary, since it contributes to reduce the risk of missing single stages in the complex management of this contingent of patients. 2. After obtaining a definitive pathohistological result and malignancy degree assessment, a revision of the complex management protocol worked out in advance is indispensable. PMID- 9854935 TI - [The diagnosis of lithiasis of the extrahepatic biliary tract]. AB - The diagnostic approach to 227 patients presenting lithiasis of the extrahepatic biliary ducts is discussed. All patients are diagnosed in the second surgical department of the University Aleksandrovska Hospital over an eleven-year period on the ground of complex examinations, including: clinical, laboratory and instrumental (echography, endoscopic retrograde cholangiopancreatography, cholangiography, computerized tomography) study. As shown by the clinical data analysis, the basic symptoms are pain--in 221 cases (97.35%), jaundice--128 (56.38%), accompanied in most of them by dyspeptic complaints, and in a minor part--by cholangitis manifestations. Of the laboratory data, the most commonly met with are the abnormally increased values of serum bilirubin, alkaline phosphatase, liver tests etc. Of the instrumental methods echography is first and foremost used in the preoperative period, supplemented in certain cases with other methods. Intraoperatively, cholangiography is routinely used for concrement detection. PMID- 9854934 TI - [Regional lymphotropic chemotherapy in breast cancer--a new approach in the therapeutic procedure for the parasternal lymph nodes]. AB - A method of regional lymphotropic chemotherapy in breast cancer is presented, aimed at attaining higher cytostatic concentration in the region of parasternal lymph nodes. In twenty-three women with centrally and medially situated tumors preoperatively 1 mg Mitoxantrone is applied around the 4 tumor poles each. In ten cases (IIb and IIIa stage) given 3 courses of nonadjuvant chemotherapy, part of the Cyclophosphamide dose is administered as regionally lymphotropic subxiphoid bilaterally. The clinical response, distribution of the Mitoxantrone blue stained regional lymph nodes and the morphological changes in the latter are analyzed. The early clinical observations indicate that the method proposed is technically easily executed, with a good local tolerance, and has a definite place in the complex treatment of breast cancer. PMID- 9854936 TI - [The Nd YAG laser in the treatment of hepatic echinococcosis]. AB - Nd YAG Laser photocoagulation for the purpose of residual cavity treatment following echinococcectomy was introduced in 1996, and practically implemented in the Department of General and Operative Surgery. Eleven patients with uncomplicated echinococcus of the hepar, 4 with purulent echinococcosis and 4 with rupture of the biliary tree, aged 26 to 80 years, are subjected to operation. The advantages and possibilities of dispersed laser application in hydatid disease treatment are discussed. The short-term results are estimated as satisfactory. PMID- 9854937 TI - [Intestinal stomas and internal splinting of the intestine in the combined treatment of peritonitis and ileus]. AB - Personal experience with the operative management of 28 patients presenting acute purulent peritonitis with ensuing ileus using a complex approach, based on total "internal" splinting of the small intestine, is shared. The 4-channel sound, designed and constructed in the clinic for intubation and simultaneous lavage of the small intestine, promotes effective staged evacuation of the toxic content from the disconnected bowel segments. What is more, a possibility is created for the formation of controlled adhesions between the intestinal loops. The significant clinical effect of bowel intubation in conjunction with adequate peritoneal cavity lavage plus infusional therapy with antibiotics and antianaerobic agents contribute greatly to the prompt recovery of patients. PMID- 9854938 TI - [The characteristics of the electrical activity of the human colon recorded noninvasively]. AB - In ten healthy volunteers and ten patients with diseases involving the upper section of the gastrointestinal tract (chronic cholecystitis, chronic gastritis) the electrical activity of the colon is registered noninvasively using skin electrodes to the projection site of sigmoid and colon descendens. Recordings are taken with original gastrograph. Apart from the electrocolograms (EColG), the changes in intracolonic pressure in the group of 10 patients are also registered. Pressure recordings (mechanogram--MG) are done in physioscript "Hellige". Two types of EColG waves are identified depending on their frequency: 1) type A waves with frequency 5.86 +/- 0.59 cpm, and 2) type B waves with frequency 2.35 +/- 0.25 cpm. Type A are low-amplitude waves and correspond to the type A waves seen in the EColG of dogs (described by the authors elsewhere), characterizing the calm period of colonic motility. When A waves appear in the EColG no changes in the mechanograms are registered. Type B are high-amplitude waves, corresponding to the B waves in dogs (described elsewhere); as shown by the mechanograms, the appearance of B waves in the EColG is followed by the appearance of high amplitude waves--the active period of colonic motility. There is a significant difference between the frequencies of either type of EColG waves and the frequency of waves in the electrogastrograms (EGG) registered synchronously with EColG. PMID- 9854939 TI - [Our experience in introducing current antibiotic prophylaxis into abdominal surgery--the initial results]. AB - Over a one-year period (Nov. 1996-Nov. 1997), a total of 154 patients are admitted on an emergency basis, with deferred emergency and for routine treatment in the Clinic of Emergency Surgery. They are distributed in three groups, as follows: patients not requiring perioperative antibiotic prophylaxis and postoperative antibiotic therapy--27, patients subjected to perioperative parenteral antibiotic prophylaxis under adequate hospital conditions--121, and patients undergoing antibiotic prophylaxis in conditions inappropriate for its application--six. The drug schemes elaborated are in conformity with worldwide and Bulgarian experience along this line, and with the concrete hospital and economical conditions in this country. In all patient indicated for antibiotic prophylaxis the listed below antibacterial agents (presented as drug schemes) are administered i.v. a single time prior to anesthesia induction: in operations on the gastrointestinal tract except for interventions in acute appendicitis: cephalotin/cefazolin 2.0 g and metronidazole 0.5 g i.v.; in operations for acute appendicitis: amoxicillin/clavulanate 1.2 g and petronidazole 0.5 f i.v.; biliary surgery free of extrahepatic cholestasis: cephalotin/cefazolin 2.0 g i.v.; biliary surgery with present or preceding extrahepatic cholestasis: cefotetan 2 g i.v.; contaminated liver cysts (parasitic and nonparasitic): cefotetan 2 g i.v.; abdominal trauma without perforation of a hollow organ: cefotetan 2 g i.v.; in plastic repair of the anterior abdominal wall (congenital defects, postoperative eventration) and in poor risk patients (local and general status: cephalotin/cefazolin 2.0 g i.v. In 135 patients the postoperative period runs a course free of complications worthy of notice. Complications directly linked to introduction of the method proposed are recorded in ten cases: operative wound suppuration (4) and hospital infection (6). Complications not related directly to the procedure are observed in nine cases: urinary tract infection (4), bronchopneumonia (2), fever with unknown source of infection (including negative hemoculture) necessitating additional antibiotic therapy (2) and secondary infection (within a week of intervention) necessitating further therapy with antibiotics. The specific features of antibiotic prophylaxis used in the various types of operative interventions are discussed from microbiological, clinical and pharmacotherapeutic viewpoints. The obtained results are compared with pertinent literature data on the issue with a special reference to the clinical efficacy attained. They mirror the approach against the background of the concrete hospital conditions in this country. The method developed is fully consistent with the level of surgical expertise in Bulgaria. All efforts should be aimed at intrahospital environment improvement by means of meticulous asepsis and antisepsis. PMID- 9854940 TI - [Infections in urology departments]. AB - The wide-spreading of intrahospital infections in urological departments on a nationwide scale are analyzed over the period 1982 through 1996. Their epidemiological, etiological and nosological characterization is also defined. In this country over 6 per cent of all hospital infections occur in urological departments. Annually, in Bulgaria 5/100 discharged urological patients develop infection in hospital conditions. Both gram-negative and gram-positive microorganisms are the etiological causing factors identified, with E. coli and Proteus mirabilis being predominant. Among the patients in urological departments urinary tract infections are the leading infectious pathology so far documented 80.85% + 0.61. The low efficacy of the prophylactic and antiepidemiological measures undertaken correlate with the low rate of diagnosing such infections, and with the insufficient theoretical and methodological training of the medical personnel along these lines. PMID- 9854941 TI - [Sclerosing cholangitis after the surgical treatment of hepatic echinococcosis]. AB - The constellation of four basic factors is taken to be the underlying cause of caustic sclerosing cholangitis development, namely: 1) scolicide agent injection into the cyst cavity, 2) cystobiliary communication, 3) p. Vateri stenosis and other causes leading to prolonged scolicide agent exposure on the biliary tree, and 4) individual sensitivity. The clinical course of the disease, its diagnosis and treatment are outlined. The prognosis of the condition is estimated as exclusively serious. PMID- 9854942 TI - [The use of anorectal manometry in the postoperative assessment of continence in congenital anorectal malformations in childhood]. AB - Objective continence evaluation in children operated for anorectal malformation gives rise to complex sociomedical and ethical problems. Its adequate solution requires the use of high-precision methods for continence capacity assessment. Anorectal manometry is one of them--it enables the adoption of clearcut criteria in estimating the effect of operation, indications for reoperation and the patient's early- and long-term prognosis. The electromanometric examination, included in the comprehensive diagnostic and therapeutic protocol, still remains a procedure indispensable in evaluating the outcome of surgery in congenital anorectal malformations. PMID- 9854943 TI - [Dieulafoy's lesion--a rare cause of gastric hemorrhage]. PMID- 9854944 TI - [The identification of tumors deriving from the sweat glands]. PMID- 9854945 TI - An unbiased errors-in-variables approach to detecting unconscious cognition. AB - Greenwald, Klinger and Schuh (1995) have proposed a regression approach for detecting unconscious cognition. An errors-in-variables approach is presented that corrects for measurement error in the predictor and takes into account that the latent predictor variable is assumed to be non-negative. The new approach requires the same input as the uncorrected regression analysis and provides consistent estimates of regression weights as well as valid statistical tests of their significance. In particular, the method yields a consistent estimate of the regression intercept that provides critical evidence for unconscious cognition. A simulation study illustrates these aspects of the new technique. Several data sets are then reanalysed by means of the new method. PMID- 9854946 TI - A comparison of three additive tree algorithms that rely on a least-squares loss criterion. AB - The performances of three additive tree algorithms which seek to minimize a least squares loss criterion were compared. The algorithms included the penalty function approach of De Soete (1983), the iterative projection strategy of Hubert & Arabie (1995) and the two-stage ADDTREE algorithm, (Corter, 1982; Sattath & Tversky, 1977). Model fit, comparability of structure, processing time and metric recovery were assessed. Results indicated that the iterative projection strategy consistently located the best-fitting tree, but also displayed a wider range and larger number of local optima. PMID- 9854948 TI - Raymond Bernard Cattell (1905-1998). PMID- 9854947 TI - Normal theory based test statistics in structural equation modelling. AB - Even though data sets in psychology are seldom normal, the statistics used to evaluate covariance structure models are typically based on the assumption of multivariate normality. Consequently, many conclusions based on normal theory methods are suspect. In this paper, we develop test statistics that can be correctly applied to the normal theory maximum likelihood estimator. We propose three new asymptotically distribution-free (ADF) test statistics that technically must yield improved behaviour in samples of realistic size, and use Monte Carlo methods to study their actual finite sample behaviour. Results indicate that there exists an ADF test statistic that also performs quite well in finite sample situations. Our analysis shows that various forms of ADF test statistics are sensitive to model degrees of freedom rather than to model complexity. A new index is proposed for evaluating whether a rescaled statistic will be robust. Recommendations are given regarding the application of each test statistic. PMID- 9854949 TI - Perinatal bereavement: the photographer's role in infant death. PMID- 9854950 TI - Three little words--vision, perception, seeing. AB - Vision, perception and seeing are everyday words, especially important to those in the image creation business. Vision, here, is used to describe the physiological process of image formation in the eye, and its subsequent projection to the brain for further action. Visual perception is a process driven by sensation with its outcome dependent on judgements based on the perceiver's situational experiences. It is a complex involving cerebral interpretations of a series of retinal images. Seeing is an intellectual exercise strongly influenced by perceptions and cultural experiences. It may be expressed in several ways, among them, verbally and pictorially. Some individuals acquire these expressions at an early age, but for most it must be learned. In the sciences, clarity is important, while in the fine arts, subtlety, and sometimes obscurity, are often sought. In all instances, the image maker is a communicator; and if the message is not received by a prepared viewer, the creator has failed. An understanding of seeing is an important mechanism in the process of image creation. An attempt will be made in this paper to advance an understanding of this process. PMID- 9854951 TI - A photographic internship in India. PMID- 9854952 TI - Subacute combined degeneration. PMID- 9854953 TI - Oscillopsia: impaired vision during motion in the absence of the vestibulo-ocular reflex. PMID- 9854954 TI - Neuropsychiatric sequelae one year after a minor head injury. PMID- 9854955 TI - Neurology and the kidney. AB - Renal failure is relatively common, but except in association with spina bifida or paraplegia it is unlikely to occur as a result of disease of the CNS. Renal failure, however, commonly affects the nervous system. The effects of kidney failure on the nervous system are more pronounced when failure is acute. In addition to the important problems related to renal failure there are both acquired and genetically determined diseases which may affect the kidney and the brain. Those acquired diseases include the vasculitides, the paraproteinaemias, and various granulomatous conditions (considered in other chapters of Neurology and Medicine). In two of the most commonly encountered genetically determined diseases, Von Hippel-Lindau disease and polycystic kidney disease, location of pathogenic mutations will provide improved screening programmes and, possibly, allow therapeutic intervention. Uraemia may affect both the central and peripheral nervous systems. Whereas the clinical features of uraemia are well documented, the pathophysiology is less well understood and probably multifactorial. Uraemic encephalopathy, which classically fluctuates, is associated with problems in cognition and memory and may progress to delirium, convulsions, and coma. The encephalopathy may initially worsen with periods of dialysis and almost certainly relates to altered metabolic states in association with ionic changes and possibly impaired synaptic function. Renal failure may affect the peripheral nervous system, resulting in a neuropathy which shows a predilection for large diameter axons. This may be reversed by dialysis and transplantation. The myopathy seen in renal failure, often associated with bone pain and tenderness, is similar to that encountered in primary hyperparathyroidism and osteomalacia. Dialysis itself is associated with neurological syndromes including the dysequilibrium syndrome, subdural haematoma, and Wernicke's encephalopathy. Dialysis dementia, which was prevalent during the 1970s, has reduced in frequency with the use of aluminium free dialysate. With the introduction of transplantation and the concomitant use of powerful immunosuppressive drugs, the pattern of neurological problems encountered in renal replacement therapy has shifted. Five per cent of patients develop nerve injuries during renal transplantation, and up to 40% of patients experience neurological side effects from cyclosporine. Furthermore, CNS infections, often fungal in type, have been reported in up to 45% of transplant patients coming to postmortem. The nature of the involvement of neurologists with their nephrology colleagues is therefore evolving. PMID- 9854956 TI - Subacute combined degeneration: clinical, electrophysiological, and magnetic resonance imaging findings. AB - OBJECTIVE: Vitamin B12 deficiency is a systemic disease that often affects the nervous system. One of the most prevalent manifestations is subacute combined degeneration (SCD) of the spinal cord. To access the clinical, electrophysiological, and structural abnormalities associated with SCD, a study was conducted in nine patients. METHODS: Clinical, electrophysiological (electroneurography, somatosensory and motor evoked potentials), and MRI evaluations were performed in patients before and after treatment. RESULTS: The most prominent clinical and electrophysiological findings in all patients were dysfunctions of the posterior column. Corresponding hyperintense lesions in the posterior column of the spinal cord were found in two patients by T2 weighted MRI. Damage to the central motor pathway was identified in four patients. Demyelinating neuropathy was present in one patient and axonal neuropathy in four. All patients showed improvement of their symptoms after treatment with cobalamin. Abnormalities of the spinal cord on MRI disappeared early in recovery. Motor evoked potentials and median somatosensory evoked potentials typically normalised after treatment, whereas tibial somatosensory evoked potentials remained abnormal in most patients. CONCLUSIONS: Clinical, electrophysiological, and MRI findings associated with SCD in vitamin B12 deficiency are diverse. Thus vitamin B12 deficiency should be considered in the differential diagnosis of all spinal cord, peripheral nerve, and neuropsychiatric disorders. PMID- 9854957 TI - Oscillopsia: visual function during motion in the absence of vestibulo-ocular reflex. AB - OBJECTIVES: To investigate (1) the effects of loss of vestibular function on spatiotemporal vision and (2) the mechanisms which enable labyrinthine defective (LD) patients to adapt to oscillopsia. METHODS: Visual function and eye movements were assessed in seven normal subjects and four LD patients with oscillopsia due to absent vestibulo-ocular reflex. Temporal vision was assessed by measurement of threshold sensitivity for detection of a target which moved across a flickering, spatially uniform background field. Spatial vision was investigated by measurements of threshold sensitivity for the detection of a target moving across a spatially modulated background in the form of square wave gratings. Velocity discrimination was assessed with drifting gratings. All measurements were made under static conditions and during oscillatory movement of either the visual stimulus or the subject (1 Hz, peak velocity 50 degrees/s). RESULTS: Temporal responses--Normal subjects and LD patients exhibited similar responses while static and under body oscillation. Spatial responses--The two groups achieved similar results under static conditions but body oscillation reduced threshold sensitivities and shifted the spatial response function towards lower spatial frequencies in the LD patients only. Similar changes in the spatial responses were seen during oscillation of the visual stimulus but these occurred in both normal subjects and LD patients. Velocity discrimination--Two LD patients achieved normal velocity discrimination but the other two showed abnormal responses to visual stimulus movement; one displayed a loss of velocity discrimination during whole body oscillation, and the other mismatched the velocity of two moving grating stimuli. CONCLUSIONS: The changes in the spatial responses are attributed to the presence of retinal slip during visual stimulus motion in all subjects or body oscillation in the LD patients. It is concluded that any visual adaptation to oscillopsia achieved by the LD patients does not influence the measured spatial response functions, which arise at an early stage of visual processing. The abnormal velocity discrimination may relate to the progressive improvement in oscillopsia reported by LD patients. PMID- 9854958 TI - Creutzfeldt-Jakob disease in Sweden. AB - OBJECTIVES: To find and investigate, retrospectively, as many cases as possible of Creutzfeldt-Jakob disease (CJD) in Sweden dying during the period 1 January 1985 to 31 December 1996 and to detect any possible case(s) of new variant CJD. METHODS: The patients were found through computer search of all death certificates in Sweden on which CJD was mentioned, through information from the Swedish neuropathologists, and spontaneous reports from Swedish doctors and hospitals. Data concerning the patients were then collected from patients' case records and from brain histopathology reports. RESULTS: In total 72 cases of spongiform encephalopathy were confirmed as definite by neuropathology, one of them with Gerstmann-Staussler-Scheinker disease. In 51 further cases there were no brain pathology data but the diagnosis "probable" (37 patients) or "possible" (14 patients) CJD according to WHO criteria could be made on clinical grounds. There was a variation in number of deaths/year, from a minimum of five (1985) to a maximum of 16 (1990). Sixty patients died during the period 1985-90 and 62 during 1991-6. The sex ratio was nearly 1:1. Calculated for a population of 8.6 million (mean of 12 years) in Sweden this gives 1.18/million/year. Age at the time of the presenting symptoms ranged from 34 to 84 years. Only one patient was under 40 at the onset of symptoms. He had a spongiform encephalopathy but prion protein staining was negative. The duration of symptoms that could be attributed to CJD was 6 months or less in 75 cases, 7-12 months in 16 cases, 1 to 2 years in 15 cases, and more than 2 years in 16 patients. By definition all patients were demented. Other more common symptoms and signs were aphasia, dysphasia, dysathria, ataxia, myoclonus, pareses of the extremities, rigidity or spasticity, different types of hyperkinesias, and other psychiatric symptoms (depression, anxiety, and aggressiveness). Less common symptoms were hallucinations (mainly visual), visual defects, sensory symptoms (paraesthesias, itching, or pain), apraxia of swallowing, and disorders of eye movements. CONCLUSIONS: The incidence, the symptomatology, the age distribution (age in years at onset and at death), and the duration of illness were similar to those of other countries except for the cases of new variant CJD in the United Kingdom. There is so far no indication of any cases of new variant CJD in Sweden. PMID- 9854959 TI - Geographical epidemiology of residence of patients with motor neuron disease in Lancashire and south Cumbria. AB - OBJECTIVES: To seek objective evidence for geographical clustering of places of residence of patients with motor neuron disease (MND). METHODS: A complete residential history from birth to onset of disease was obtained from a cohort of 130 patients with MND from Lancashire and south Cumbria presenting to the Department of Neurology in Preston between 1 January 1989 and 31 December 1993. These data were compared with population based reference data from the 1991 UK Census. RESULTS: Some areal units showed a greater, others a lesser, number of MND patient residences than expected. The results suggest that the background population incidence of MND is relatively low and that the overall incidence figures previously quoted have been skewed upwards by areas in which the incidence of MND is relatively increased. These findings were further tested by Poisson modelling. The Poisson model provided a poor fit for the data at postcode district and sector levels confirming that patients with MND were significantly more likely to have lived in some areas than others after allowing for variation in population of the different areal units and for variation in duration of residence. CONCLUSIONS: These findings reinforce the results of previous work, much of which has been qualitative rather than quantitative. The results presented here suggest a low background incidence of MND in the context of generally quoted overall incidence figures. This low background incidence is, however, skewed upwards by some areal units with a relatively high incidence, thus achieving overall incidence rates comparable with generally quoted figures. We conclude that there is prima facie evidence of spatial patterns in the distribution of places of residence of patients with MND. Further examination of occupational and environmental factors in the lives of the patients with MND is required to obtain a better understanding of the importance of these findings. PMID- 9854960 TI - Clinical and electrophysiological evaluation of dysphagia in myasthenia gravis. AB - OBJECTIVE: To evaluate dysphagia at the oropharyngeal stage of swallowing and to determine the pathophysiological mechanisms of dysphagia in patients with myasthenia gravis. METHODS: Fifteen patients with myasthenia gravis with dysphagia and 10 patients without dysphagia were investigated by a combined electrophysiological and mechanical method described previously. Laryngeal movements were detected by a piezoelectric transducer and the related submental EMG (SM-EMG) and sometimes the EMG of cricopharyngeal muscle of the upper esophageal sphincter (CP-EMG) were recorded during dry or wet swallowing. The results of these electrophysiological variables were compared with those of normal age matched control subjects. RESULTS: In patients with myasthenia gravis with dysphagia, it was found that the time necessary for the larynx to remain in its superior position during swallowing and swallowing variability in successive swallows increased significantly compared with normal subjects and with patients with myasthenia gravis without dysphagia. The total duration of SM-EMG activity was also prolonged in both groups but more severely in the dysphagic patients. Electromyographic activity of the CP sphincter was found to be normal in the dysphagic patients investigated. All the patients with myasthenia gravis with dysphagia had pathological dysphagia limits (<20 ml water) whereas other patients except two, were within normal limits. CONCLUSIONS: Because the electrophysiological variables related to oropharyngeal swallowing were prolonged even in patients with myasthenia gravis without dysphagia, it is concluded that the submental and laryngeal elevators are involved subclinically in myasthenia gravis and, because of compensating mechanisms, the patient may not be dysphagic. As the CP-EMG behaviour was found to be normal, a coordination disorder between normal CP sphincter muscle and the affected striated muscles of the laryngeal elevators may be one of the reasons for dysphagia in myasthenia gravis. This method also made it possible to investigate the myasthenic involvement in the laryngeal elevators that cannot be evaluated by other electrophysiological methods in myasthenia gravis. PMID- 9854962 TI - The structural and functional mechanisms of motor recovery: complementary use of diffusion tensor and functional magnetic resonance imaging in a traumatic injury of the internal capsule. AB - OBJECTIVES: Recovery from focal motor pathway lesions may be associated with a functional reorganisation of cortical motor areas. Previous studies of the relation between structural brain damage and the functional consequences have employed MRI and CT, which provide limited structural information. The recent development of diffusion tensor imaging (DTI) now provides quantitative measures of fibre tract integrity and orientation. The objective was to use DTI and functional MRI (fMRI) to determine the mechanisms underlying the excellent recovery found after a penetrating injury to the right capsular region. METHODS: DTI and fMRI were performed on the patient described; DTI was performed on five normal controls. RESULTS: The injury resulted in a left hemiplegia which resolved fully over several weeks. When studied 18 months later there was no pyramidal weakness, a mild hemidystonia, and sensory disturbance. fMRI activation maps showed contralateral primary and supplementary motor cortex activation during tapping of each hand; smaller ipsilateral primary motor areas were activated by the recovered hand only. DTI disclosed preserved structural integrity and orientation in the posterior capsular limb by contrast with the disrupted structure in the anterior limb on the injured side. CONCLUSIONS: The findings suggest that the main recovery mechanism was a preservation of the integrity and orientation of pyramidal tract fibres. The fMRI studies do not suggest substantial reorganisation of the motor cortex, although ipsilateral pathways may have contributed to the recovery. The initial deficit was probably due to reversible local factors including oedema and mass effect; permanent damage to fibre tracts in the anterior capsular limb may account for the persistent sensory deficit. This study shows for the first time the potential value of combining fMRI and DTI together to investigate mechanisms of recovery and persistent deficit in an individual patient. PMID- 9854961 TI - Morvan's fibrillary chorea: a paraneoplastic manifestation of thymoma. AB - Morvan's fibrillary chorea is a rare disease characterised by symptoms which include neuromyotonia, cramping, weakness, pruritus, hyperhidrosis, insomnia, and delirium. The first case of Morvan's fibrillary chorea to be associated with clinical manifestations of myasthenia gravis with thymoma, psoriasis, and atopic dermatitis is reported. Muscle histopathology disclosed chronic denervation and myopathic changes and in vitro electrophysiology demonstrated both presynaptic and postsynaptic defects in neuromuscular transmission. Serum antibodies to acetylcholine receptors, titin, N-type calcium channels, and voltage gated potassium channels were detected. Plasmapheresis, thymectomy, and long term immunosuppression induced a dramatic resolution of symptoms. The association of thymoma with other autoimmune disorders and autoantibodies, and prolonged and sustained remission with chronic immunosuppression, place Morvan's fibrillary chorea on the range of neurological diseases arising as a paraneoplastic complication of cortical thymomas. PMID- 9854963 TI - Cognitive effects of pituitary tumours and their treatments: two case studies and an investigation of 90 patients. AB - OBJECTIVES: Two case studies are reported of patients with pituitary adenomas who had been treated with trans-sphenoidal surgery, one with and one without adjunctive radiotherapy, in whom memory impairment was found. Further to this, neuropsychological investigations of 90 patients were carried out (1) to establish the prevalence of such deficits, and (2) to try to determine their cause. METHODS: Two case studies are described. For the expanded study, patients were recruited from the data base of the endocrinology department of St Thomas's Hospital, London, if they had previously been treated for a pituitary adenoma in the past 30 years. Ninety patients were contacted and assessed with a wide range of neuropsychological tests. They were divided into five treatment groups: those who had received transfrontal surgery with radiotherapy, trans-sphenoidal surgery with or without radiotherapy, radiotherapy only, and a bromocriptine therapy group, as well as a group of 19 healthy control subjects matched for age and sex. RESULTS: In the two patients presented, both showed severe memory impairments compared with their intact intellectual ability. The more severely affected patient had received adjunctive radiotherapy, and superimposition of the 90% isodose fields on a postoperative MRI examination suggested involvement of the diencephalic structures. In the group study, significant deficits in anterograde memory were also obtained on two measures (WMS-R, RMT) for all patient groups when compared with the healthy controls, although these impairments varied in degree and were less in the bromocriptine group. However, the individual surgical and radiotherapy treatment groups did not differ significantly from one another. By contrast, general intellectual function (IQ) remained intact for all groups, as did performance on supplementary cognitive tests, including measures of frontal lobe or "executive" function, language comprehension, and speed of mental processing. Psychiatric morbidity and tumour aetiology did not seem to relate to the presence of memory deficits. CONCLUSIONS: Anterograde memory deficits were seen in the two case studies and in all our treatment groups when compared with the healthy controls, and these occurred in the context of preserved intellectual function. The present findings suggest that these memory deficits result from treatment rather than from the underlying tumour, but there was no difference between the effects of surgery and radiotherapy. It is suggested that they result from damage to diencephalic structures implicated in memory. PMID- 9854964 TI - Sex and relationship dysfunction in neurological disability. AB - OBJECTIVES: (1) to ascertain how many people with neurological disability experience sexual or relationship problems; (2) to examine the interplay of neurological disability and sexual function within the context of the dyadic relationship; (3) to consider the implications of the results for service provision. METHODS: A survey of outpatients attending Hunters Moor Regional Rehabilitation Centre, Newcastle upon Tyne over a 6 month period. Standardised measures of sexual function and concern, relationship satisfaction, disability, and mental health were used. RESULTS: Seventy people were interviewed (18% of the potential study population). Prevalence figures therefore refer to this self selected sample. Fifty one per cent had experienced a change in sexual function and 27% were concerned about this change. Of those in cohabiting relationships 25% were experiencing difficulties. Gender was significantly associated with concern about sexual function, men being more concerned. A high score on the marital adjustment scale indicating relationship dissatisfaction was the best predictor of change in and concern about sexual function. Change in sexual function and duration of illness were the most powerful predictors of relationship dissatisfaction. CONCLUSION: If concern is taken as an indication of a desire for help more than one in four of this sample required help for sexual dysfunction. Sexual and relationship functioning were very closely associated and dysfunction in either of these areas was the best predictor of dysfunction in the other area. The dyadic relationship is an important institution in the management of disability. Sexual dysfunction can be predictive of difficulties within a relationship. Any service designed to address sexual health should also address relationship issues. PMID- 9854965 TI - Dementia and aphasia in motor neuron disease: an underrecognised association? AB - OBJECTIVES: To determine the prevalence and nature of global cognitive dysfunction and language deficits in an unselected population based cohort of patients with motor neuron disease (MND). METHODS: A battery of neuropsychological and language tests was administered to patients presenting consecutively over a 3 year period to a regional neurology service with a new diagnosis of sporadic motor neuron disease. RESULTS: The 18 patients could be divided on the basis of their performance into three groups: Three patients were demented and had impaired language function (group 1); two non-demented patients had an aphasic syndrome characterised by word finding difficulties and anomia (group 2). Major cognitive deficits were therefore found in five of the 18 patients (28%). The remaining 13 performed normally on the test battery apart from decreased verbal fluency (group 3). CONCLUSIONS: The prevalence of cognitive impairment in MND in this population based study of an unselected cohort was higher than has been previously reported. Language deficits, especially anomia, may be relatively frequent in the MND population. Aphasia in MND may be masked by dysarthria and missed if not specifically examined. PMID- 9854966 TI - Forgetting rates in neuropsychiatric disorders. AB - OBJECTIVE: Previous studies have attributed accelerated forgetting rates on recognition memory tasks to temporal lobe pathology, but findings in some patient groups may have been attributable to metabolic disruption. Findings in psychiatric disorders such as schizophrenia are conflicting. The purpose of the present study was to compare forgetting rates in patients with confusional states (post-electroconvulsive therapy (post-ECT), delirium), with those obtained in schizophrenic patients (with putative temporal lobe pathology), non-ECT depressed patients, and healthy controls. The findings could also be compared with previous reports in patients with head injury, focal structural lesions, and Alzheimer's dementia. METHODS: Two studies employed a picture recognition task to examine forgetting rates, the first between delays of 1 minute, 15 minutes, and 30 minutes, and the second between delays of 10 minutes, 2 hours, and 24 hours. RESULTS: There were no significant differences in forgetting rates between 1 minute and 30 minutes, but the ECT group showed accelerated forgetting between 10 minutes and 2 hours compared with healthy controls, associated with a rapid decline in "hit rate". This was not attributable to differential changes in either depression or severity of memory impairment. There were no differences in forgetting rates across the other subject groups. CONCLUSION: Post-ECT confusional state patients (similarly to "within post-traumatic amnesia" patients with head injury) show accelerated forgetting on a recognition memory task and, in this, they contrast with patients who have focal structural lesions or widespread cortical atrophy. Accelerated forgetting may reflect the effect of disrupted cerebral metabolism on either "consolidation" or memory "binding" processes. PMID- 9854967 TI - Neuropsychiatric sequelae one year after a minor head injury. AB - OBJECTIVE: To assess neuropsychiatric sequelae 1 year after minor head injury in a cross sectional study using home interviews with patients and their relatives at 1 year after head injury. METHODS: The study cohort included 148 adults who were admitted to hospital after a minor head injury between 1 July 1994 and 30 June 1995 and showed clinical or radiological evidence of brain injury. Main outcome measures used in the study were the Glasgow outcome scale, Edinburgh rehabilitation status scale, Barthel index, clinical interview schedule-revised, mini mental state examination, and assessment of symptoms of postconcussional syndrome. RESULTS: At one year follow up, four (2.9%) patients had a severe disability, 35 (25.5%) had a moderate disability, and 95 (69.3%) had no disability according to the Glasgow outcome scale. A slightly higher proportion (33.3%, n=45) showed disability according to the Edinburgh rehabilitation status scale. Thirty one patients (23.1%) scored <24 in the mini mental state examination. These were mostly patients over the age of 65. Twenty three patients (17.2%) were diagnosed as psychiatric cases according to the clinical interview schedule-revised scale. Seventy four (55.2%) patients showed one of the symptoms of postconcussional syndrome. The most commonly shown neurobehavioural problems were irritability (30%), sleep disturbance (29%), and impatience (27%). CONCLUSION: One year after a minor head injury, a substantial proportion of patients showed neuropsychiatric sequelae. PMID- 9854968 TI - Selective sparing of topographical memory. AB - The case of a 61 year old patient with Pick's disease involving predominantly the left temporal lobe, who has been studied over a 5 year period, is reported. She presented with a grave impairment of both verbal and non-verbal memory functions. Her non-verbal memory deficits included profound impairments on the recognition of unfamiliar faces and the recall of abstract designs. Remarkably, her visual recognition memory performance for unknown buildings, landscapes, and outdoor scenes was preserved. Strikingly, her ability to recall familiar routes and learn new ones through a complex virtual reality town was also entirely normal. This seems to be the first case documenting the selective preservation of topographical memory in the context of severe non-verbal and verbal memory impairments. These findings imply that topographical memory and non-verbal memory are subserved by separable neural systems. PMID- 9854969 TI - Sex differences in patients with mesial temporal lobe epilepsy. AB - Possible sex differences in the pattern of interictal hypometabolism were investigated, and also seizure spread in patients with mesial temporal lobe epilepsy (n=48) and hippocampal sclerosis (MTLE). Male patients (n=21) more often had a frontal lobe hypometabolism ipsilateral to the seizure onset (p<0.0001) and a spread of epileptiform activity to this region (p=0.001). By contrast, female patients more often exhibited hypometabolism (p=0.0052) and an ictal spread to the contralateral temporal lobe (p=0.0097). These findings suggest sex differences in spatial distribution of brain dysfunction in MTLE, perhaps reflecting sexual dimorphism in regional cerebral connectivity. PMID- 9854970 TI - Regional metabolism: associations with dyscalculia in Alzheimer's disease. AB - OBJECTIVES: The ability to calculate, which is an important aspect of social daily living, is commonly impaired in patients with Alzheimer's disease even early in the course of the disease. Dyscalculia is often accompanied by focal brain damage, and has been argued to be an independent sign localised around the left temporoparietal region. However, the region most responsible for dyscalculia in Alzheimer's disease has not been determined. The relation between calculation ability and regional cerebral glucose metabolism in Alzheimer's disease was therefore examined. METHODS: The calculation ability, In 91 patients with probable Alzheimer's disease of minimal to moderate severity, was assessed using the arithmetic subtest of the Wechsler adult intelligence scale-revised and the performance correlated with regional cerebral glucose metabolism determined by "F fluorodeoxyglucose and PET. RESULTS: Regional glucose metabolism in the left inferior parietal lobule and in the left inferior temporal gyrus was significantly correlated with the calculation performance irrespective of age, sex, education, and severity of disease. CONCLUSIONS: The results suggest that dysfunction of the left inferior parietal lobule and the left inferior temporal gyrus plays an important part in producing dyscalculia in patients with Alzheimer's disease. PMID- 9854971 TI - Jean Baptiste van Helmont (1577-1644). PMID- 9854972 TI - Case report of unusual leukoencephalopathy preceding primary CNS lymphoma. AB - A previously healthy 35 year old woman presented with bilateral uveitis associated with multiple, evolving, non-enhancing white matter lesions consistent with a progressive leukoencephalopathy such as multiple sclerosis. Thirty months after her initial presentation, she was diagnosed with primary CNS lymphoma and died 14 months later. The unusual clinical course preceding the diagnosis suggests that a demyelinating disease may have preceded, and possibly heralded, the development of primary CNS lymphoma. Cases of "sentinel lesions" heralding the diagnosis of primary CNS lymphoma have been reported, and this case further corroborates such instances and raises further issues regarding possible neoplastic transformation occurring in inflammatory diseases such as multiple sclerosis. PMID- 9854973 TI - Time resolution for visual information processing in Parkinson's disease. AB - It has been suggested that a deficit in timing could be the cause of the sensory disturbances reported for Parkinson's disease. To test this hypothesis the temporal discrimination thresholds in four visual tasks were used to study 45 healthy young people, 14 healthy elderly people, and 17 patients with Parkinson's disease. In these tasks, subjects watched a computer controlled light emitting diode display and pushed a button when the visual event previously specified by the researcher was perceived. The time between successive images required to discriminate a visual detail was accurately quantified. In two of the four tasks, the time for visual processing of image sequences was longer in the elderly group than in the young group. No significant differences were found between patients with Parkinson's disease and their age matched controls for any of the four tasks. Present data show normal temporal discrimination and no slowing in the initial steps of visual processing in Parkinson's disease. PMID- 9854974 TI - Depletion of neutralising antibodies resensitises a secondary non-responder to botulinum A neurotoxin. AB - The objective was to evaluate whether removal of neutralising antibodies potentially resensitises a secondary non-responder to botulinum neurotoxin A (BoNT/A). Neutralising antibodies directed against BoNT/A are produced during long term treatment with BoNT/A-hemagglutinin complex in up to 10% of patients with cervical dystonia. These patients become secondary non-responders. Other serotypes of BoNT are not yet generally available and may also bear the risk of inducing antibody formation. Plasma exchange (PE) (one treatment cycle) and immunoadsorption on a protein A column (IA-PA; three treatment cycles) was employed over 15 months to remove neutralising antibodies from a severely disabled secondary non-responder with cervical dystonia. After plasma exchange or IA-PA, BoNT/A was reinjected. Antibodies were measured with a sensitive functional toxin neutralising test. Repeated use of plasma exchange and IA-PA depleted neutralising antibodies to below the detection limit and subsequently allowed successful BoNT/A injection into dystonic muscles. No serious side effects were found related to the depletion of IgG. In conclusion PE or IA-PA performed before BoNT/A readministration may provide an alternative strategy in treating selected secondary non-responders who are severely disabled. PMID- 9854975 TI - Psychological associations with emotionalism after stroke. AB - The psychological associations with emotionalism were examined, 1 month after stroke, in 448 stroke survivors who met inclusion criteria for a randomised trial of psychological treatment. One hundred and one (21.5%) patients had emotionalism. Thirty eight (38%) of those with emotionalism had a clinically significant mood disorder, compared with 64 of 347 (18%) of those without emotionalism. Emotionalism was associated with 10 other psychiatric syndromes, the presence of most of which could be accounted for by the coexistence of depression with emotionalism. In a logistic regression analysis, irritability and ideas of reference were associated with emotionalism after adjustment for the presence of depression. These psychological associations with emotionalism may be incorporated into a psychological model of emotionalism as a disorder of emotional control, and they may help in the understanding of the socially disabling effects of this common condition. PMID- 9854976 TI - Jugular venous and arterial concentrations of serum S-100B protein in patients with severe head injury: a pilot study. AB - The objective of this study was to analyse the temporal course of the jugular venous-arterial gradient of S-100B protein after severe head injury and the correlation between the absolute concentrations of serum S-100B protein and outcome, CT findings, and clinical variables. Fifteen patients were included in this pilot study. All patients were treated according to a standard therapy protocol targeted to maintain cerebral perfusion pressure. The serum concentration of S-100 protein was measured daily for five consecutive days after injury by a monoclonal two site immunoluminometric assay. Nine patients showed favourable and six unfavourable outcome after 6 months with a mortality rate of 33% (five patients). The mean gradient between jugular venous and arterial blood was 8.2% (p<0.05). Patients showing an unfavourable outcome had significantly higher jugular venous or arterial S-100 values compared with those with a favourable outcome (jugular venous S-100B 2.78 microg/l v 1.22 microg/l, p<0.05; arterial S-100B 2.48 microg/l v 1.19 microg/l, p<0.05). All patients with an initial or secondary increase in S-100B value of >2 microg/l were found to have an unfavourable outcome. S-100B was found to be an independent predictor of outcome after severe head injury. The persisting increase of S-100B for three to five days even in patients with favourable outcome and no signs of secondary insults might reflect continuing damage to the blood-brain barrier or ongoing glial cell death. PMID- 9854977 TI - Behavioural disturbance and visual hallucinations in a 78 year old man. PMID- 9854978 TI - Metastatic choriocarcinoma presenting as multiple intracerebral haemorrhages: the role of imaging in the elucidation of the pathology. PMID- 9854979 TI - Systematic review of antifibrinolytic treatment in aneurysmal subarachnoid haemorrhage. PMID- 9854981 TI - High resolution magnetic resonance imaging in peripheral vestibular dysfunction in multiple sclerosis. PMID- 9854980 TI - Wallerian degeneration of the pyramidal tract after a thrombotic stroke. PMID- 9854982 TI - Vertebral aneurysm due to isolated spontaneous dissection of the intracranial vertebral artery. PMID- 9854983 TI - A novel de novo point mutation in the GTP cyclohydrolase I gene in a Japanese patient with hereditary progressive and dopa responsive dystonia. PMID- 9854984 TI - Central nervous system involvement in four patients with Charcot-Marie-Tooth disease with connexin 32 extracellular mutations. PMID- 9854985 TI - Lack of mutation G209A in the alpha-synuclein gene in French patients with familial and sporadic Parkinson's disease. PMID- 9854986 TI - Resolution of extensive spinal epidural haematoma with conservative treatment. PMID- 9854987 TI - Flail arm syndrome: a distinctive variant of amyotrophic lateral sclerosis. PMID- 9854988 TI - Multifocal cortical myoclonus and cerebral amyloid beta-peptide angiopathy. PMID- 9854989 TI - Antiganglioside antibodies in various episodes in a patient with recurrent Guillain-Barre syndrome. PMID- 9854990 TI - Urinary retention associated with a unilateral lesion in the dorsolateral tegmentum of the rostral pons. PMID- 9854991 TI - Splicing of the glutamate transporter EAAT2: a candidate gene of amyotrophic lateral sclerosis. PMID- 9854992 TI - Lhermitte's sign in cavernous angioma of the cervical spinal cord. PMID- 9854993 TI - Autosomal dominant paroxysmal kinesigenic choreoathetosis: a clinical and genetic study of two families. PMID- 9854994 TI - Granulomatous angiitis of the CNS causing spontaneous intracerebral haemorrhage: the importance of leptomeningeal biopsy. PMID- 9854995 TI - Intravesical atropine suppression of detrusor hyperreflexia in multiple sclerosis. PMID- 9854996 TI - Prothrombotic mutations and ischaemic stroke at a young age in two sisters. PMID- 9854997 TI - Molecular analysis of lacI mutants from transgenic fibroblasts exposed to 1,2 epoxybutene. AB - 1,3-Butadiene (BD) is a genotoxic carcinogen that is bioactivated to at least two mutagenic metabolites: 1,2-epoxybutene (EB) and 1,2:3,4-diepoxybutane (DEB). We reported previously that lacI transgenic mice exposed to BD had an increased frequency of specific base substitution mutations in the bone marrow and spleen relative to unexposed controls. In the experiments described here, we determined the mutagenicity and mutational spectrum of EB in Rat2 lacI transgenic fibroblasts as a means of assessing the contribution of this metabolite to the lacI mutational spectrum of BD. Rat2 cells were exposed to 0, 0.4, 0.6, 0.8 or 1.0 mM EB for 24 h, resulting in a range of cell survival from 100 to 15%, respectively. Mutagenicity was assessed at 0, 0.6 and 1.0 mM EB. Unexposed controls had a background mutant frequency of 6 +/- 1 +/- 10(-5), while the mutant frequency in cells exposed to 0.6 and 1.0 mM EB was increased 2- and 3 fold, respectively. DNA sequence analysis of 154 lacI mutants recovered in these experiments revealed an increase in the frequency of specific base substitution mutations in cells exposed to 1.0 mM EB compared with controls. These included G:C-->A:T transitions at non-CpG sites, G:C-->T:A transversions and A:T-->T:A transversions, which have all been observed in lacI mutants isolated from transgenic mice exposed to BD. These results suggest that EB causes mutation primarily by base substitution and that the spectrum of these mutations closely resembles that of BD. These data, along with previous findings from our laboratory, suggest that EB is more likely than DEB to be primarily responsible for the lacI mutational spectrum observed in lacI transgenic mice exposed to BD. PMID- 9854998 TI - Suppression of tumorigenicity of human lung carcinoma cells after transfection with connexin43. AB - The human lung carcinoma cell line PG is defective in gap junctional intercellular communication (GJIC). Connexin43 (Cx43) mRNA, which is expressed in normal human lung cells, is undetectable in these tumor cells. To explore if up regulation of Cx43 gene expression will suppress malignancy of PG cells, Cx43 cDNA was co-transfected with pSV2neo cDNA into PG cells. Control cells were transfected with the blank vector plus neo cDNA. Several stable Cx43 transfectant clones, which acquired high levels of Cx43 expression and the capacity of GJIC, were compared with control clones and the parental cell line, both of which lacked Cx43 expression and GJIC. The control clones resembled the parental cells in exhibiting high cell growth rate, weak attachment to the substratum and a high frequency of colony formation in soft agar. In contrast to the control cells, Cx43 transfected clones showed reduced growth rate, enhanced attachment to the substratum and inhibition of colony formation in soft agar. In vivo results from nude mice experiments showed high tumorigenicity with control clones and inhibition of tumorigenicity in Cx43 transfected clones. The consistency between in vitro and in vivo results strongly suggests a tumor suppressing effect of the Cx43 gene in human lung carcinoma cells. PMID- 9854999 TI - Melatonin does not inhibit estradiol-stimulated proliferation in MCF-7 and BG-1 cells. AB - Melatonin, an indolic pineal hormone, is produced primarily at night in mammals and is important in controlling biological rhythms. Previous research suggested that melatonin can attenuate proliferation in the estrogen-responsive MCF-7 breast cancer cell line. We tested whether these anti-proliferative effects may have physiological consequences upon two estrogen-responsive cell lines, MCF-7 (a breast cancer cell line) and BG-1 (an ovarian adenocarcinoma cell line). Melatonin (10(-9)-10(-5) M) attenuated proliferation of MCF-7 and BG-1 cells by >20% in the absence of estrogen. However, 17beta-estradiol exposure negated the ability of melatonin to inhibit proliferation. To substantiate this finding, cells were estrogen starved followed by multiple treatments with estradiol and melatonin. Melatonin did not inhibit estradiol-stimulated proliferation under this protocol. Estradiol increased MCF-7 and BG-1 cell cycle transition from G1 to S phase, however, melatonin did not inhibit this transition nor did it down regulate estradiol-induced pS2 mRNA levels measured by northern blotting, further indicating that melatonin was unable to attenuate estradiol-induced proliferation and gene expression. We also examined the effects of melatonin on estradiol induced proliferation in MCF-7 cell xenografts in athymic nude mice. Melatonin at a dose 28 times greater than 17beta-estradiol did not inhibit estradiol-induced proliferation in vivo. Furthermore, pinealectomy did not increase proliferation. Therefore, we conclude that melatonin does not directly inhibit estradiol-induced proliferation. PMID- 9855000 TI - Estrogen-responsive RING finger mRNA induction in gastrointestinal carcinoma cells following bile acid treatment. AB - The underlying molecular mechanisms of the tumor-promoting activity of bile acids such as chenodeoxycholic acid (CDCA) and deoxycholic acid (DCA) and the protective effect of ursodeoxycholic acid (UDCA) remain largely unclear. Using RNA arbitrarily primed PCR (RAP-PCR) for differential display, we identified, cloned and sequenced differentially expressed transcripts after treating gastric carcinoma cells (St 23132) with the bile acids CDCA, DCA and UDCA. One of these transcripts was identified to be an estrogen-responsive RING finger protein (efp) mRNA. The differential expression of efp in gastric cancer cells was confirmed by low stringency RT-PCR. efp mRNA levels were induced 3-fold in gastric carcinoma cells after CDCA and DCA treatment, whereas no change in expression was detected after UDCA treatment. Finally, treatment of the colon carcinoma cell line HT 29 with DCA resulted in a 2- to 5-fold induction of efp mRNA levels whereas UDCA did not induce efp. As expected, efp expression was also increased after 24 h of estrogen treatment. In summary, a synergy or a common pathway of tumor enhancement of bile acids and estrogen via efp in gastrointestinal carcinogenesis can be envisioned. PMID- 9855001 TI - Isolation and characterization of a spontaneously transformed malignant mouse mammary epithelial cell line in culture. AB - A method is described that permits the selection of spontaneously transformed mammary epithelial colonies from an untransformed mouse mammary epithelial cell line, NMuMG, and utilizes a long-term anchorage-independent growth of the transformants on soft agarose. These transformed cells (NMuMG-ST) are shown to be distinguishable from the untransformed cells by morphology, growth characteristics, induced carcinomas when transplanted into nude mice and ability to metastasize. This transformed phenotype displayed focal, multilayer growth and higher saturation density in comparison with the untransformed phenotype. Transplanted tumors as well as metastatic lung tumors in nude mice were adenocarcinomas morphologically similar to typical mammary tumors in humans. This selection procedure of mutant mammary cells from an immortalized cell line derived from normal mammary glands could be very useful to identify the genomic biomarkers in the growth regulation and malignant progression of breast cancer. PMID- 9855002 TI - Adaptive protection against the induction of oxidative DNA damage after hyperbaric oxygen treatment. AB - Hyperbaric oxygen (HBO) treatment (i.e. exposure to 100% oxygen at a pressure of 2.5 ATA for a total of three 20 min periods) of human subjects caused clear and reproducible DNA effects in the comet assay with leukocytes. Interestingly, DNA damage was detected only after the first treatment and not after further treatments under the same conditions, indicating an increase in antioxidant defences. We now demonstrate that blood taken 24 h after HBO treatment is well protected against the in vitro induction of DNA damage by hydrogen peroxide (H2O2). H2O2 treatment caused a significant induction of DNA effects in the comet assay and chromosome breakage in the micronucleus test in the blood of volunteers before HBO. The same treatment did not cause genotoxic effects 24 h after HBO. This protective effect lasted for at least 1 week. Experiments with isolated lymphocytes gave similar results, indicating that the adaptive response is a cellular effect. The cells were not comparably protected against the genotoxic effects of gamma-irradiation, suggesting increased scavenging of reactive oxygen species distant from nuclear DNA. PMID- 9855003 TI - Determination of malondialdehyde-induced DNA damage in human tissues using an immunoslot blot assay. AB - Malondialdehyde (MDA) is a product of lipid peroxidation and prostaglandin biosynthesis. It is mutagenic and carcinogenic and the major adduct formed by reaction with DNA, a highly fluorescent pyrimidopurinone (M1-dG), has been detected in healthy human liver and leukocyte DNA. Analytical methods used so far for the detection of M1-dG have not been applied to a large number of individuals or variety of samples. Often, only a few microg of DNA from human tissues are available for analysis and a very sensitive assay is needed in order to detect background levels of M1-dG in very small amounts of DNA. In this paper, the development of an immunoslot blot (ISB) assay for the measurement of MI-dG in 1 microg of DNA is described. The limit of detection of the assay is 2.5 adducts per 10(8) bases. A series of human samples were analysed and levels of 5.6-9.5 (n = 8) and 3.1-64.3 (n = 42) of M1-dG per 10(8) normal bases were detected in white blood cell and gastric biopsy DNA, respectively. Results on four human samples were compared with those obtained using an HPLC/32P-post-labelling (HPLC/PPL) method previously developed and indicated a high correlation between M1-dG levels measured by the two assays. The advantages of ISB over other assays including HPLC/PPL, such as the possibility of analysing 1 microg DNA/sample and the fact that it is less time-consuming and laborious, means that it can be more easily used for routine analysis of a large number of samples in biomonitoring studies. PMID- 9855004 TI - Allelic losses and DNA methylation at DNA mismatch repair loci in sporadic colorectal cancer. AB - Normal and tumor DNA samples of 35 patients with sporadic colorectal carcinoma were analyzed for microsatellite alterations at 12 markers linked to mismatch repair loci: hMLH1, hMSH2, hMSH3, hMSH6, hPMS1 and hPMS2. Remarkably, no correlation was observed between the replication error phenotype (RER+) and allelic losses at these loci. Hemizygous deletions, seen in 6/35 (17%) informative cases at hMLH1, 4/27 (15%) at hMSH2/hMSH6 and 6/34 (18%) at hMSH3, were rarely found in RER+ tumors. Since mismatch repair protein components act in molecular complexes of defined stoichiometry we propose that hemizygous deletion of the corresponding loci may be involved in colorectal tumorigenesis through defects in cellular functions other than replication error correction. The analysis of the methylation status of the promoter region of hMLH1 revealed that methylation might be an important mechanism of this locus inactivation in RER+ sporadic colorectal cancer. PMID- 9855005 TI - Resistance to 6-thioguanine in mismatch repair-deficient human cancer cell lines correlates with an increase in induced mutations at the HPRT locus. AB - Although the resistance to the cytotoxic response of certain DNA damaging agents has been well characterized in cells deficient in mismatch repair, little is known about how such resistance affects mutagenesis. Using human cancer cell lines defective in mismatch repair (MMR) and complementary cell lines in which the MMR defects were corrected by chromosome transfer, we present the cytotoxic effect and the mutagenic response at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus following exposure to the chemotherapeutic agent, 6 thioguanine (6-TG). Upon exposure to 6-TG, there was a differential cytotoxic response. The MMR-deficient cells were resistant to 6-TG exposure up to 5 microM, whereas the MMR-proficient cell lines were significantly more sensitive at the same levels of exposure. Furthermore, the mutagenic response at HPRT induced by 6 TG was substantially increased in the MMR-deficient lines relative to the MMR proficient cell lines. These findings support the notion that cytotoxicity to 6 TG is mediated through functional MMR and that resistance to the cytotoxic effects of 6-TG is directly associated with an increase in induced mutations in MMR-defective cells. These data suggest that the use of 6-TG as a chemotherapeutic agent may result in the selection of MMR-defective cells, thereby predisposing the patient to an increased risk for developing secondary tumors. PMID- 9855006 TI - Suppressive effects of nimesulide, a selective inhibitor of cyclooxygenase-2, on azoxymethane-induced colon carcinogenesis in mice. AB - The effects of nimesulide, a selective inhibitor of cyclooxygenase-2 (COX-2) on azoxymethane (AOM)-induced colon carcinogenesis were investigated in mice. AOM at a dose of 10 mg/kg body wt was administered to male ICR mice once a week for 6 weeks. The animals were fed on AIN-76A powder diet containing nimesulide at doses of 200 or 400 p.p.m., starting the day before the first carcinogen treatment until the end of the experiment, at week 30. Administration of nimesulide reduced the incidence of colon carcinomas to 32 and 25% for the AOM + 200 and 400 p.p.m. nimesulide groups, respectively, compared with the AOM + basal diet group (50%). Multiplicities of colon carcinomas in the 200 and 400 p.p.m. nimesulide-treated groups were 0.70 +/- 0.28 and 0.35 +/- 0.11, respectively, being significantly smaller than the AOM alone value (1.79 +/- 0.47). The sizes of the colon carcinomas in the nimesulide-treated groups were also decreased. No significant influence on liver and lung tumor development was apparent. Thus, nimesulide exerted a suppressive effect on AOM-induced colon carcinogenesis in mice. PMID- 9855007 TI - Catechol-O-methyltransferase and breast cancer risk. AB - Recent studies suggest that a polymorphism in catechol-O-methyltransferase (COMT) is associated with increased risk of breast cancer. Methylation by COMT is the principal pathway for inactivation of catechol estrogens, which are hypothesized to participate in estrogen-induced carcinogenesis. We examined the association of COMT genotype and breast cancer risk in a population-based, case-control study of invasive breast cancer in North Carolina. The study population consisted of 654 cases and 642 controls, with approximately equal numbers of African-American and white women and women under the age of 50 and aged 50 or over. Contrary to previous reports, we did not observe an association between one or more copies of the low activity COMT allele (COMT-L) and breast cancer risk. Multivariate relative risks (RRs) were 0.8 (95% confidence interval: 0.6-1.1) for COMT-HL and 0.8 (0.6-1.1) for COMT-LL, compared with the COMT-HH genotype. RRs for COMT did not differ among African-American and white women and we did not observe strong modification of RR estimates by menopausal status, body mass index, physical activity or other covariates. Our results suggest that COMT genotype is not related to breast cancer risk. PMID- 9855009 TI - Increased aneusomy and long arm deletion of chromosomes 5 and 7 in the lymphocytes of Chinese workers exposed to benzene. AB - Two of the most common cytogenetic changes in therapy- and chemical-related leukemia are the loss and long (q) arm deletion of chromosomes 5 and 7. The detection of these aberrations in lymphocytes of individuals exposed to potential leukemogens may serve as useful biomarkers of increased leukemia risk. We have used a novel fluorescence in situ hybridization (FISH) procedure to determine if specific aberrations in chromosomes 1, 5 and 7 occur at an elevated rate in the blood cells of workers exposed to benzene. Forty-three healthy workers exposed to a wide range of benzene concentrations (median 31 p.p.m., 8 h time-weighted average) and 44 unexposed controls from Shanghai were studied. Whole blood was cultured and metaphase spreads were harvested at 72 h. Benzene exposure was associated with increases in the rates of monosomy 5 and 7 but not monosomy 1 (P < 0.001, P < 0.0001 and P = 0.94, respectively) and with increases in trisomy and tetrasomy frequencies of all three chromosomes. Long arm deletion of chromosomes 5 and 7 was increased in a dose-dependent fashion (P = 0.014 and P < 0.0001) up to 3.5-fold in the exposed workers. These results demonstrate that leukemia specific changes in chromosomes 5 and 7 can be detected by FISH in the peripheral blood of otherwise healthy benzene-exposed workers. We suggest that aberrations in chromosomes 5 and 7 may be useful biomarkers of early biological effect for benzene exposure. PMID- 9855008 TI - Associations between both genetic and environmental biomarkers and lung cancer: evidence of a greater risk of lung cancer in women smokers. AB - This molecular epidemiologic case-control study of lung cancer incorporated three complementary biomarkers: the glutathione S-transferase M1 (GSTM1) null genotype, a potential marker of susceptibility, and polycyclic aromatic hydrocarbon-DNA adducts (PAH-DNA) and sister chromatid exchanges (SCE), both indicators of environmentally induced genetic damage. Associations between biomarkers and lung cancer were investigated, as were possible gene-environment interactions between the GSTM1 null genotype and tobacco smoke exposure. Subjects included 136 primary non-small cell lung cancer surgical patients and 115 controls at the Columbia Presbyterian Medical Center. Questionnaire and Tumor Registry data, pre-treatment blood samples and biomarker measurements on blood were obtained. Overall, GSTM1 null genotype was significantly associated with lung cancer [odds ratio (OR) = 2.04, 95% confidence interval (CI) = 1.13-3.68]. ORs for GSTM1 and lung cancer were significant in females (2.50, 1.09-5.72) and smokers (2.25, 1.11-4.54) and not significant in males (1.4, 0.58-3.38) and non-smokers (0.88, 0.18-4.33). However, ORs for males versus females and smokers versus non-smokers did not differ significantly. The OR for GSTM1 and lung cancer in female smokers was 3.03 (1.09-8.40), compared with 1.42 (0.53-4.06) in male smokers. In contrast to PAH DNA adducts in leukocytes, SCE did not differ between cases and controls. Neither biomarker differed significantly between the two GSTM1 genotypes. The combined effect of elevated PAH-DNA adducts and GSTM1 genotype on case-control status (16.19, 1.2-115) appeared multiplicative. Results suggest that the effect of the GSTM1 null genotype is greatest in female smokers, which is consistent with other evidence that indicates that women are at higher risk of lung cancer than males, given equal smoking. Persons with both the GSTM1 deletion and elevated PAH-DNA adducts may represent a sensitive subpopulation with respect to carcinogens in tobacco smoke and other environmental media. PMID- 9855010 TI - A study of multiple biomarkers in coke oven workers--a cross-sectional study in China. AB - We conducted a cross-sectional molecular epidemiological study of coke oven workers exposed to the established carcinogen polycyclic aromatic hydrocarbons (PAHs) to evaluate the relationships between both traditional 'exposure markers' and a series of biomarkers, including urinary 1-hydroxypyrene as a marker of internal dose, leukocyte aromatic DNA adducts as markers of biologically effective dose, serum p53 protein as a response marker and genetic polymorphisms of cytochrome P4501A1 and glutathione S-transferase MI as susceptibility markers. Twenty-five male subjects each were randomly selected from the top, middle and bottom work areas of the oven, and the control plant. They were matched for age and smoking status. The mean levels of PAH exposure, monitored by stationary and personal samplers, and of worker urinary 1-hydroxypyrene differed significantly between the top, middle and bottom of the oven and control work areas. The highest stationary and personal PAH concentrations and 1-hydroxypyrene levels were demonstrated at the top work area. Good correlations were found between the stationary PAH levels, personal PAH levels and urinary 1-hydroxypyrene levels. No positive correlations were demonstrated between aromatic DNA adduct levels and current or cumulative PAH exposure dose. In the presence of genetic polymorphisms of cytochrome P4501A1, a positive correlation was demonstrated between aromatic DNA adducts and urinary 1-hydroxypyrene levels. There was also a significant correlation between serum p53 protein levels and the cumulated benzo[a]pyrene exposure dose. Although these biomarkers have certain limitations, they are applicable to cancer epidemiology, and may contribute to our understanding of the mechanisms of carcinogenesis. PMID- 9855011 TI - Metabolism of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by human cytochrome P4501A1, P4501A2 and P4501B1. AB - While the metabolic activation of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) by N-hydroxylation has been well documented, the relative roles of the human cytochrome P450 (CYP) enzymes that catalyze this reaction have not been established. Previous studies indicated that the mutagenic activation product, 2 hydroxyamino-PhIP (N2-OH-PhIP), is produced primarily by CYP1A2, and to a lesser extent by CYP1A1. We recently reported that human CYP1B1 also produces N2-OH-PhIP (Carcinogenesis, 18, 1793-1798, 1997). In the present study, we examined PhIP metabolism by microsomes containing recombinant human CYP1A1, 1A2 or 1B1 expressed in Sf9 insect cells and compared the kinetic values for PhIP metabolite formation. PhIP metabolites were analyzed by high pressure liquid chromatography with fluorescence and absorbance detection. Vmax values for N2-OH-PhIP formation were 90, 16 and 0.2 nmol/min/nmol P450, and the apparent Km values were 79, 5.1 and 4.5 microM for human CYP1A2, 1A1 and 1B1, respectively. The non-mutagenic metabolite, 4'-hydroxy-PhIP, was also formed by all three CYP enzymes with Vmax values of 1.5, 7.8 and 0.3 nmol/ min/nmol P450 and apparent Km values of 43, 8.2 and 2.2 microM for human CYP1A2, 1A1 and 1B1, respectively. Although the Vmax for N2-OH-PhIP production was highest for CYP1A2, the catalytic efficiency (Vmax/Km) of CYP1A1 was greater than that of CYP1A2. These results suggest that, for humans, extrahepatic CYP1A1 may be more important than previously thought for the metabolic activation of the dietary carcinogen PhIP. PMID- 9855012 TI - Detoxification of optically active bay- and fjord-region polycyclic aromatic hydrocarbon dihydrodiol epoxides by human glutathione transferase P1-1 expressed in Chinese hamster V79 cells. AB - Dihydrodiol epoxides (DEs) are important carcinogenic metabolites of polycyclic aromatic hydrocarbons (PAHs). The metabolic formation of four stereoisomeric DEs (a pair of optically active diastereomers termed as syn- and anti-form) is possible. Glutathione tranferases (GSTs) have been demonstrated to catalyze the detoxification of DEs. Purified GSTs display remarkable differences in catalytic efficiencies towards bay- and fjord-region DEs along with a high degree of regio- and stereoselectivity. Here we determined to which extent heterologously expressed human GSTP1-1, a major GST isoform in lung, affects the mutagenicity of stereoisomeric bay-region DEs of benzo[a]pyrene in Chinese hamster V79 cells. To evaluate the influence of sterical crowding in the substrate on the activity of GSTP-1, the study was extended to the strongly mutagenic fjord-region (-)-anti DEs of benzo[c]phenanthrene and dibenzo[a,l]pyrene. GSTP1-1,reduced preferentially the mutagenicity (studied at the hprt locus) of (+)-anti and (+) syn-DEs of benzo[a]pyrene (by 66 and 67%) as compared with the corresponding (-) anti- and (-)-syn-enantiomers (by 15 and 13%). These results are in line with previous studies on the enantioselectivity of purified GSTP1-1 towards the DE isomers of benzo[a]pyrene and benzo[c]phenanthrene showing that enantiomers with (R)-configuration at the benzylic oxiranyl carbon are better substrates than those with (S)-configuration. Interestingly, the (-)-anti-DEs of benzo[c]phenanthrene and dibenzo[a,l]pyrene were efficiently detoxified by GSTP-1 1 in the constructed cell line (reduction of mutagenicity by 66 and 64%). This study demonstrates that differences in the caalytic activity seen for purified GST towards individual mutagens do not necessarily reflect the detoxification of DEs by the same enzyme in a living cell and provides further evidence that specific human GSTs play a role in the detoxification of DEs of PAHs. PMID- 9855013 TI - Sex differences in oxidative damage in ddY mouse kidney treated with a renal carcinogen, iron nitrilotriacetate. AB - Iron-induced free radical injuries in male and female ddY mice, especially the sex difference and its mechanisms, were studied after an i.p. injection of a renal carcinogen, ferric nitrilotriacetate. Male mice were much more susceptible to iron-induced free radical injuries than female mice. Oxidative modification of proteins and DNA occurred more strongly in males than in females, as measured by protein carbonyl content and 8-hydroxydeoxyguanosine, respectively. Histochemical detection of 4-hydroxy-2-nonenal-modified proteins using an antibody and DNA fragmentation as detected by the TUNEL method also showed that males are more severely damaged than females, especially in the proximal convoluted tubules. These results could not be explained by the difference in iron status between male and female mice. In fact, the toxic so-called 'free' iron in serum and kidney were not different between male and female mice and storage iron, such as ferritin and hemosiderin, was also comparable in both kidneys. In previous studies we proposed the glutathione cycling hypothesis to explain the sex differences. The half-life of glutathione in the kidney was significantly shorter in males (29 min) than in females (57 min), as determined by the glutathione decrease after buthionine sulfoximine treatment, a specific inhibitor of glutathione synthesis. The specific activity of gamma-glutamyltranspeptidase (EC 2.3.2.2) in female mice was 73% of that in male mice. These results suggest that the faster glutathione turnover in males could account for the higher susceptibility to oxidative injury by supplying the reducing equivalent that reduces Fe(III) to Fe(II), thereby facilitating iron-catalyzed free radical reactions. PMID- 9855015 TI - Structural determination of a mutagenic aminophenylnorharman produced by the co mutagen norharman with aniline. AB - Norharman (9H-pyrido[3,4-b]indole), widely distributed in our environment, including cigarette smoke and cooked foodstuffs, is not mutagenic to Salmonella strains, but becomes mutagenic to S.typhimurium TA98 and YG1024 with S9 mix in the presence of non-mutagenic aromatic amines such as aniline and o-toluidine. To elucidate the mechanisms of co-mutagenicity, we tried to isolate the mutagen(s) produced by a reaction between norharman and aniline with S9 mix. By HPLC purification, two mutagenic compounds (I and II), one (I) showing mutagenicity with and the other (II) without S9 mix, were isolated. The structure of compound I was deduced to be a coupled compound of norharman and aniline, 9-(4' aminophenyl)-9H-pyrido[3,4-b]indole (aminophenylnorharman), by a variety of spectrometry techniques and this was confirmed by its chemical synthesis. The mutagenic activity of this novel heterocyclic amine was tested using the pre incubation method and was found to induce 187,000 revertants in TA98 and 1,783,000 revertants in YG1024 per microg with S9 mix. Compound II was shown to be hydroxyaminophenylnorharman. Formation of the same DNA adducts was observed in YG1024 when aminophenylnorharman or a mixture of norharman plus aniline was incubated with S9 mix. The hydroxyamino derivative also yielded the same DNA adducts in YG1024. Thus, the appearance of mutagenicity by norharman with aniline in the presence of S9 mix suggests that the coupled mutagenic compound, aminophenylnorharman, is formed from norharman and aniline, then converted to the hydroxyamino derivative and forms DNA adducts to induce mutations in TA98 and YG1024. PMID- 9855014 TI - Role of peroxisome proliferator-activated receptor alpha in altered cell cycle regulation in mouse liver. AB - The mechanisms underlying peroxisome proliferator-induced hepatocarcinogenesis are unclear but are mediated by the peroxisome proliferator-activated receptor alpha (PPARalpha). To determine the role of PPARalpha in the mechanisms of hepatocarcinogenesis, the effect of Wy-14,643 on expression patterns of acyl CoA oxidase (ACO) and proteins involved in cell proliferation in the PPARalpha-null mouse were evaluated. ACO, CDK-1, CDK-2, CDK-4, PCNA and c-myc proteins were significantly increased in wild-type mice fed Wy-14,643 for 5 weeks or 11 months, as compared with controls. This effect was not observed in Wy-14,643-treated PPARalpha-null mice. Expression patterns of cyclin B1, cyclin D, cyclin E and p53 were not different in any of the groups. mRNAs encoding CDK-1, CDK-4, cyclin D1 and c-myc were also increased in wild-type mice fed Wy-14,643 but not in PPARalpha-null mice. These results indicate that the increase in CDK-1, CDK-4 and c-myc may be caused by an increase in transcription that is mediated directly or indirectly by PPARalpha. Thus PPARalpha-dependent alterations in cell cycle regulatory proteins induced by peroxisome proliferators are likely to contribute to the hepatocarcinogenicity of peroxisome proliferators. PMID- 9855016 TI - Altered expression of cyclin D1 and cyclin-dependent kinase 4 in azoxymethane induced mouse colon tumorigenesis. AB - Alterations in the expression of the cell cycle regulators, cyclin D1 and cyclin dependent kinase 4 (Cdk4), have been implicated in malignancies of both humans and experimental animal models. We hypothesize that altered expression of cyclin D1 and Cdk4 may also be involved in mouse colon tumorigenesis induced by the chemical carcinogen, azoxymethane (AOM). In the present study, SWR/J mice were given AOM by i.p. injection at a dose of 10 mg/kg once a week for 8 weeks, and colonic tissue and tumors were isolated 18 weeks later. The expression and localization of cyclin D1 and Cdk4 were examined by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical analyses. Cyclin D1 and Cdk4 mRNA levels in tumor samples were increased 1.3-fold (P < 0.01) and 1.2 fold (P < 0.01), respectively, when compared with control mouse colon tissue. Control colon epithelium was uniformly negative for cyclin D1 immunoreactivity, whereas minimal Cdk4 nuclear staining was confined to the lower portion of the crypts within the control tissue. Both cyclin D1 and Cdk4 immunoreactive cells were markedly increased in preneoplastic lesions and in adenomas isolated from AOM-treated mice. Furthermore, some morphologically normal colon crypts from AOM treated mice showed positive cyclin D1 immunoreactivity. These findings suggest that overexpression of cyclin D1 and Cdk4 occurs early in the AOM-induced mouse colon tumorigenesis and may contribute to tumor progression in this model. PMID- 9855017 TI - Sulfation of alpha-hydroxytamoxifen catalyzed by human hydroxysteroid sulfotransferase results in tamoxifen-DNA adducts. AB - The formation of tamoxifen (TAM)-derived DNA adducts was investigated by incubation of DNA with (E)-alpha-hydroxytamoxifen [(E)-alpha-OHTAM], 3' phosphoadenosine 5'-phosphosulfate (PAPS), and human recombinant sulfotransferase. Using 32P-post-labeling and HPLC analysis, two TAM-DNA adducts were detected in incubations that included the human hydroxysteroid sulfotransferase SULT2A1 (hHST). When compared with standards of stereoisomers of alpha-(N2-deoxyguanosinyl)tamoxifen 3'-monophosphate (dG3'P-N2-TAM), the major adduct was identified chromatographically as an epimer of the transform of dG-N2 TAM, and the minor adduct was identified as an epimer of the cis-form. The amount of TAM adducts formed by hHST was approximately three times less than that formed by an equivalent amount of rat hydroxysteroid (alcohol) sulfotransferase a. These results indicate that sulfation of alpha-OHTAM catalyzed by hHST results in the formation of dG-N2-TAMs, highly miscoding lesions, in human tissues. PMID- 9855018 TI - The duration of phorbol-inducible ErbB2 tyrosine dephosphorylation parallels that of receptor endocytosis rather than threonine-686 phosphorylation: implications for the physiological role of protein kinase C in growth factor receptor signalling. AB - Tumour cell growth may be accelerated by protein kinase C (PKC) agonists such as phorbol esters and receptor tyrosine kinases, but receptor tyrosine kinases are in turn desensitized to growth factors by PKC agonists. To clarify this apparent PKC bifunctionality, we have used phosphoantibodies to determine the relationship between PKC-dependent phosphorylation events affecting the ErbB2 oncoprotein in G8/DHFR 3T3 cells. Neither the kinetics nor the extent of phorbol-induced juxtamembrane domain (Thr686) phosphorylation vary directly with C-terminal (Tyr1222) dephosphorylation, with Tyr1222 continuing to be dephosphorylated long after Thr686 phosphorylation has also declined. Platelet-derived growth factor (PDGF) mimics the short-term effects of phorbol on Thr686 and Tyr1222 phosphorylation, and confocal microscopy reveals that both of these PKC agonists induce rapid internalization of PKC-modified ErbB2. Phorbol causes sustained cytoplasmic accumulation of PKC-phosphorylated receptors, however, whereas PDGF triggers the appearance of this ErbB2 subset only briefly. Metabolic labelling and co-precipitation studies fail to implicate heterologous molecules in either the tyrosine dephosphorylation or internalization of PKC-modified ErbB2. Taken in the context of earlier juxtamembrane domain mutagenesis studies, these findings indicate that phorbol-activated PKC may desensitize growth factor receptors to extracellular ligands solely by triggering sustained receptor internalization. We submit that PKC-dependent juxtamembrane domain phosphorylation represents a physiological mechanism for shortening the duration and enhancing the specificity of growth factor signalling by promoting internalization of liganded and unliganded receptors, respectively. PMID- 9855019 TI - Evaluation of the in vitro direct and indirect genotoxic effects of cobalt compounds using the alkaline comet assay. Influence of interdonor and interexperimental variability. AB - The mechanisms of cobalt-induced pulmonary interstitial fibrosis and cancer are incompletely understood. DNA damage, either induced by genotoxic (direct or via oxygen radicals) or co-genotoxic (e.g. inhibition of DNA repair) processes may play an important role in the initiation of cancer. The alkaline comet assay provides a sensitive tool to investigate these two processes. Cobalt metal, a mixture of cobalt with tungsten carbide and cobalt chloride, were compared for their DNA-damaging capacity. Concentrations from 0 to 6.0 microg Co-equivalent/ml were tested. All three compounds were able to induce DNA damage in isolated human lymphocytes from three donors, in a dose- and time-dependent way. A relatively large interexperimental and interdonor variability in response was observed. This was ascribed to technical parameters and unidentified individual factors. This confirms the importance of repeating experiments using the same and different donors. The DNA-damaging potential of the cobalt-tungsten carbide mixture was higher than that of cobalt metal and cobalt chloride, which had comparable responses. No significant increase of DNA migration was observed when the DNA of cells treated with cobalt metal, cobalt-tungsten carbide or tungsten carbide were incubated with the oxidative lesion-specific enzyme formamidopyrimidine DNA glycosylase. This suggests that during the short treatment period no substantial oxidative damage to DNA was produced. Cobalt metal was able to inhibit the repair of methylmethanesulphonate-induced DNA damage. This was concluded from simultaneous exposure to cobalt and methyl methanesulphonate, post-incubation and post-treatment with 1.2 microg/ml cobalt of methyl methanesulphonate-treated cells. PMID- 9855020 TI - Acetaldehyde alters proliferation, differentiation and adhesion properties of human colon adenocarcinoma cell line Caco-2. AB - Studies with experimental animals indicate that acetaldehyde, the first metabolite of ethanol that is microbially formed in the colonic lumen, may play a role in ethanol-associated colorectal co-carcinogenesis. Although intracoIonic acetaldehyde concentrations are highest during the metabolism of exogenous ethanol, some individuals may also possess marked amounts of endogenous acetaldehyde. Since no information is available concerning the possible effects of acetaldehyde on human colonic epithelial cells, this study was aimed to assess whether this compound, either alone or in combination with ethanol, affects such properties of human neoplastic colonocytes that are considered relevant with regard to cancer development. Human colon adenocarcinoma cell line Caco-2 was used as a model of transformed colonocytes, and effects of acetaldehyde and/or ethanol on the proliferation and differentiation of these cells as well as on their adhesion to collagens I and IV, the most important extracellular matrix proteins in the colon, were studied. The results of this study show that acetaldehyde markedly affects the phenotype of Caco-2 cells without having direct cytotoxic effects. Like many carcinogens, it was found to have a dual effect on cell proliferation rate, acute exposure being inhibitory and chronic exposure stimulating. Acetaldehyde also considerably decreased both sucrase activity and nuclear content of protein kinase A catalytic subunit in Caco-2 cells, which indicate that the differentiation of the cells was disturbed. Moreover, the adhesion of Caco-2 cells to collagens I and IV was dose-dependently reduced by acetaldehyde treatment. All these changes, i.e. enhanced cell proliferation rate (by chronic treatment), decreased differentiation, and reduced adhesion to extracellular matrix proteins, would in vivo predict more aggressive and invasive tumour behaviour. The possibility that colonic intraluminal acetaldehyde, either ethanol-derived or endogenous, might enhance the development of colorectal tumours should therefore be considered. PMID- 9855021 TI - Influence of aromatic hydrocarbon receptor-mediated events on the genotoxicity of cigarette smoke condensate. AB - The role of aromatic hydrocarbon receptor (AhR)-mediated events on the genotoxicity of mainstream cigarette smoke condensate was investigated. In vitro studies with mouse hepatoma cells stably transfected with a DRE-dependent luciferase reporter indicate that cigarette smoke condensate is able to transform AhR to an active form which is capable of initiating gene transcription. Micronucleus formation in two hepatoma cell lines was used as an index of genotoxicity. Cigarette smoke condensate was observed to induce a higher frequency of micronuclei in Hepa1c1c7 cells relative to TAOc1BP(r)c1 cells, which express approximately 10-fold less AhR. Furthermore, the frequency of micronuclei was potentiated when Hepa1c1c7 cells were pretreated with 2,3,7,8 tetrachlorodibenzo-p-dioxin, a high affinity ligand of AhR. These in vitro studies were followed by an in vivo experiment with Ahr+/+ and Ahr-/- mice. Animals were dosed for three consecutive days with cigarette smoke condensate (0.5-10 microg/kg/day, i.p. injection). The frequency of micronuclei in reticulocytes and total erythrocytes was determined in peripheral blood samples collected 24 h after the last administration. While condensate was found to increase the incidence of micronucleated reticulocytes in Ahr+/+ mice, no increase was observed in the null allele animals. Furthermore, the frequency of micronucleated erythrocytes, a measure of basal chromosome-damaging activity, was slightly but significantly higher in Ahr+/+ relative to Ahr-/- mice. Together, these data suggest that cigarette smoke contains chemicals which transform the AhR to an active transcription factor and AhR-regulated enzyme induction plays an important role in mediating the genotoxicity of this complex environmental pollutant. PMID- 9855022 TI - Estrogen treatment enhances hereditary renal tumor development in Eker rats. AB - Hormonal influences are known to affect the development of renal cell carcinoma in man and laboratory animal models. We tested the hypothesis that estrogen treatment or ovariectomy of rats modulates renal tumor development using tuberous sclerosis 2 (Tsc2) heterozygous mutant (Eker) rats in which a germline mutation predisposes the animals to renal cell tumor development. Two-month-old female wild-type and Eker rats were ovariectomized or sham-operated and treated with placebo or 5 mg 17beta-estradiol in s.c. pellets for 6 or 10 months. Rats were examined at 8 or 12 months of age, at which time the numbers of renal tumors and preneoplastic foci were quantitated and the severity of nephropathy was assessed. In contrast to what may have been expected, prolonged estrogen treatment enhanced the development of hereditary renal cell tumors, with a 2-fold greater number of preneoplastic and neoplastic renal lesions compared with untreated Eker rats. Ovariectomized Eker rats had 33% fewer renal lesions than the unmanipulated control group. No tumors or preneoplastic lesions were present in wild-type rats at either time point. Estrogen treatment increased the severity of nephropathy in both wild-type and Eker rats, whereas ovariectomy was protective against nephropathic changes. Although estrogen is not a rat renal carcinogen, it enhanced the development of hereditary renal cell tumors when administered to Eker rats. Eker rats heterozygous for a mutation in the Tsc2 locus provide a good model in which to study how genetic and hormonal factors contribute to the development of renal cell tumors and to understand the influence genetic susceptibility has on the development of renal cell carcinoma. PMID- 9855023 TI - Bioactivation of the cooked food mutagen N-hydroxy-2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine by estrogen sulfotransferase in cultured human mammary epithelial cells. AB - Cooked food mutagens from fried meat and fish have recently been suggested to contribute to the etiology of breast cancer. Thus, the most prevalent of these compounds, i.e. 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, or rather its more mutagenic N-hydroxylated metabolite (N-OH-PhIP), forms DNA adducts in mammary cells, including human mammary epithelial (HME) cells. The objective of this study was to determine the involvement of estrogen sulfotransferase (EST), the only sulfotransferase identified in HME cells, in the further bioactivation of N-OH-PhIP. These studies were done in vitro using human recombinant EST and in intact HME cells. Human recombinant EST increased the covalent binding of [3H]N OH-PhIP to calf thymus DNA approximately 3.5-fold in the presence of the sulfotransferase co-substrate 3'-phosphoadenosine-5'-phosphosulfate at each N-OH PhIP concentration (1, 10 and 100 microM) (n = 6, P < 0.001). In contrast, EST did not catalyze the DNA binding of two other cooked food mutagens, N-hydroxy-2 amino-3-methylimidazo[4,5-f]quinoline and N-hydroxy-2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline, which are mainly hepatocarcinogens. Cultured HME cells displayed high EST activity, which could be completely inhibited by 1 microM estrone. When the cells were incubated with [3H]N-OH-PhIP, binding to native DNA occurred at 60-240 pmol/mg DNA. This binding was inhibited to 55% of control by 1 microM estrone (P < 0.01, n = 8), suggesting that EST plays a significant role in carcinogen bioactivation in human breast tissue. PMID- 9855024 TI - Effects of dietary anticarcinogens on rat gastrointestinal glutathione S transferase theta 1-1 levels. AB - Several naturally occurring food components or non-steroidal anti-inflammatory drugs (NSAIDs) may reduce gastrointestinal cancer rates. Recently we have shown that dietary administration of such compounds enhanced the glutathione S transferase (GST) enzyme activity and class alpha, mu and pi isoenzyme levels in the rat gastrointestinal tract. Elevation of the levels of GSTs, a family of biotransformation enzymes with many functions such as detoxification of carcinogens, might be one of the mechanisms that lead to cancer prevention. We therefore investigated whether the anticarcinogens alpha-angelicalactone, alpha tocopherol, beta-carotene, coumarin, ellagic acid, flavone, indole-3-carbinol, d limonene, oltipraz, phenethylisothiocyanate (PEITC) and the sulphoraphane analogue compound-30 affect gastrointestinal rGSTT1-1 protein levels in male Wistar rats. rGSTT1-1 protein levels were determined in cytosolic fractions of liver and oesophageal-, gastric-, small intestinal- and colonic mucosa by densitometrical analyses of western blots after immunodetection with an anti human GSTT1-1 monoclonal antibody, that cross-reacts with rGSTT1-1. In control Wistar rats, gastrointestinal rGSTT1-1 protein levels were highest in the liver and decreased in the order liver > stomach > colon > oesophagus > small intestine. Gastric rGSTT1-1 protein levels were enhanced by alpha angelicalactone, alpha-tocopherol, coumarin, ellagic acid, oltipraz, PEITC and the sulphoraphane analogue compound-30. Oesophageal rGSTT1-1 protein levels were elevated by a-angelicalactone and coumarin, whereas colonic rGSTT1-1 protein levels were elevated by coumarin. Ellagic acid, on the other hand, reduced hepatic rGSTT1-1 protein levels to 53% of the control. In conclusion, dietary anticarcinogens are capable of inducing rGSTT1-1 protein levels in the rat gastrointestinal tract, and are most pronounced in the stomach. Enhanced rGSTT1-1 protein levels might lead to an increase of enzyme activity and to a more efficient detoxification of carcinogens and thus could contribute to prevention of carcinogenesis. PMID- 9855025 TI - Angiotensin II receptor blockade and end-organ protection: pharmacological rationale and evidence. AB - BACKGROUND: The renin-angiotensin system is a widely studied hormonal system that comprises substrate-enzyme interactions, the end result of which is the production of the active peptide angiotensin II. Because angiotensin II affects blood pressure control, sodium and water homeostasis, and cardiovascular function and structure, a great deal of research effort has been directed toward blocking the renin-angiotensin system. Angiotensin II also may be involved in end-organ damage in hypertension, heart failure, and vascular disease. ANGIOTENSIN II RECEPTORS: At least two subtypes of angiotensin II receptors have been identified, angiotensin type 1 (AT)1 and type 2 (AT2). The AT1 receptor mediates all the known actions of angiotensin II on blood pressure control. Additionally, research has indicated that the AT1 receptor modulates cardiac contractility and glomerular filtration, increases renal tubular sodium reabsorption, and cardiac and vascular hypertrophy. Less is known about the function of the AT2 receptor. Evidence suggests that the AT2 receptor inhibits cell proliferation and reverses the AT1-induced hypertrophy. Indeed, these receptors are thought to exert opposing effects. ANGIOTENSIN RECEPTOR ANTAGONISTS: This newly introduced class of drugs is able to inhibit the renin-angiotensin system at the receptor level by specifically blocking the AT1 receptor subtype. These drugs induce a dose dependent blockade of angiotensin II effects, resulting in reduced blood pressure, urinary protein, and glomerular sclerosis. It is postulated that AT1 receptor antagonists may provide end-organ protection by blocking angiotensin II effects via the AT1 receptor, leaving the AT2 receptor unopposed. Consequently, these agents may reduce the morbidity and mortality that result from myocardial infarction and other conditions resulting from structural alterations in the heart, kidney, and vasculature. PMID- 9855026 TI - A look through the new therapeutic window: irbesartan. AB - Although an impressive array of efficacious antihypertensive agents are available to treat hypertension, the optimal use of these agents is limited by dose-related side-effect profiles. This is particularly the case for widely used first-line antihypertensive agents such as diuretics, beta-blockers, calcium antagonists, and alpha1-blockers; this represents a major therapeutic dilemma in treating hypertension. With the development of the angiotensin II receptor antagonists (AIIRAs), this dilemma might have been solved. Irbesartan is a long-acting AIIRA that provides dose-related efficacy with placebo-like tolerability at all clinical doses. The results of placebo and active-control trials of irbesartan have demonstrated that the agent is as effective as the leading members of major antihypertensive classes with respect to blood pressure control, while having superior tolerability. Pooled data from nine multicenter, randomized, placebo controlled trials with irbesartan have documented no adverse events caused by dose-response. This feature could widen the traditionally narrow therapeutic window in the treatment of hypertension and point to the use of AIIRAs such as irbesartan as first-line therapy in the management of hypertension. PMID- 9855027 TI - Optimizing antihypertensive therapy in patients with diabetic nephropathy. AB - The number of cases of diabetic nephropathy is increasing, especially among patients with non-insulin-dependent diabetes mellitus (NIDDM). It is difficult to prevent the occurrence or progression of NIDDM, and current levels of treatment are below standard. According to one study, actuarial 5-year survival rates are only about 38% for patients with insulin-dependent diabetes mellitus and 9% for those with NIDDM receiving renal replacement therapy. Because cardiovascular diseases are responsible for more than half of these deaths, hypertension, as a major contributing factor to cardiac death, is a crucial component in the therapy for such patients. It is well established that lowering blood pressure is an important preventive measure to be taken in patients with diabetic nephropathy; blockade of the renin-angiotensin system offers benefits beyond lowering blood pressure in type I diabetic nephropathy. Two major trials are currently underway to determine the effects of angiotensin II receptor antagonists on nephropathy in patients with NIDDM. The Irbesartan Diabetic Nephropathy Trial (IDNT) has already enrolled approximately 85% of the proposed 1650 NIDDM patients to be randomly assigned to placebo, irbesartan or amlodipine. Baseline characteristics of the initial cohort are presented. In light of the well-documented case for blockade of the renin-angiotensin system in diabetes, the potentially superior blockade afforded by angiotensin II receptor antagonists, and the superior tolerability of these agents, trials such as the IDNT take on special importance for the treatment of diabetic patients. From these data may come the justification for the belief that angiotensin II receptor antagonists impart greater benefits in the treatment of diabetes than merely their well-documented role in lowering blood pressure. PMID- 9855028 TI - The importance of left ventricular hypertrophy in human hypertension. AB - Hemodynamic and non-hemodynamic factors contribute to the development of left ventricular hypertrophy (LVH). The presence of LVH is an important independent risk factor for total mortality and for cardiovascular morbidity and mortality. Direct cardiac effects of LVH include an increased risk of developing of congestive heart failure, an increased risk of arrhythmic events, and a reduced coronary flow reserve, promoting myocardial ischemic episodes. In addition, hypertension may promote the development of coronary artery atherosclerosis. The prognostic implications of LVH underscore the importance of diagnostic procedures. The electrocardiogram has a high specificity to identify patients with LVH but the sensitivity is fairly low. Echocardiography provides higher sensitivity and also gives important information, such as the pattern of left ventricular geometry, which is of prognostic importance, and the presence of diastolic dysfunction, which is an early abnormality in the evolution of hypertensive LVH. Reversal of LVH appears to improve prognosis. Reduction of blood pressure is one important component in the regression of LVH. Important quantitative differences exist between drug classes in the reversal of cardiac hypertrophy despite similar antihypertensive effects, suggesting other factors to be of importance in the regression of left ventricular mass. LVH is reduced more by angiotensin-converting enzyme inhibitors than by other antihypertensive drug classes, suggesting an effect on structural myocardial changes beyond that provided by the reduction of blood pressure. Recent data suggest that angiotensin II receptor antagonists (AIIRAs) have quantitatively similar effects on left ventricular mass as do angiotensin-converting enzyme inhibitors. A comparative trial of the AIIRA irbesartan and the beta-blocker atenolol demonstrated that despite similar reductions in blood pressure, the reductions attained in left ventricular mass with irbesartan were progressive and numerically greater than those attained with atenolol. Taken together, these findings provide circumstantial evidence for an important role of angiotensin II acting on angiotensin type 1 (AT1) receptors in the development or maintenance of cardiac hypertrophy. Confirmation of the favorable effects of angiotensin-converting enzyme inhibitors and AIIRAs on left ventricular mass in larger trials, including those assessing cardiovascular morbidity and mortality, will be of major importance in the future treatment of hypertension. PMID- 9855029 TI - Existing treatment strategies: does noncompliance make a difference? AB - Although published literature shows a clear relationship between poor patient compliance with antihypertensive medication and failure to achieve blood pressure control, the association between treatment choices and compliance is less clear. Patient behavior in randomized controlled trials differs from that in actual practice; studies of this difference are therefore required to consider real world evidence. Studies of actual practice have used a variety of measures of patient behavior, which makes comparisons difficult. In addition, they often have mingled patients receiving antihypertensive treatment for the first time and established hypertensive patients--groups that generally have quite different levels of compliance. This article briefly reviews the literature concerning therapeutic choices and patient compliance with hypertensive medications, including a recent study of persistence using treatment data from the provincial health plan of Saskatchewan, Canada. The Saskatchewan analysis was a rigorous examination of actual practice treatment for hypertension, which demonstrated that angiotensin-converting enzyme inhibitors were associated with the highest 1- and 4.5-year persistence rates and the lowest discontinuation rates among calcium antagonists, beta-blockers and diuretics. These findings may hold important implications for physicians when choosing an initial antihypertensive therapy, and point to the potential importance of the excellent safety and tolerability profiles of new antihypertensive therapies. PMID- 9855030 TI - The use of generalized tests in medical research. PMID- 9855031 TI - Conceptual and statistical issues in the validation of analytic dilution assays for pharmaceutical applications. AB - The discovery, research, and development of a pharmaceutical product relies on the availability of validated assays for assessing product characteristics and drug effects in vivo and in vitro. Development of a validated assay is a multifaceted activity that provides many interesting challenges for bioanalytical chemists and statisticians. In this paper, the similarity condition for fundamental validity of an analytic dilution assay is reviewed as a basic concept underlying the validation of assays for pharmaceutical applications. The distinction between the validity and the acceptability of an assay is considered in terms of the characteristics evaluated during four stages of validation. Recent guidelines on the validation of analytical procedures published by the U.S. Food and Drug Administration are appraised from a statistical perspective, and statistical issues in the validation process are discussed. PMID- 9855032 TI - The conditional binomial test revisited for clinical trials. AB - McNemar's test is used to test the hypothesis that one treatment is better than another in a matched-pair design for binary outcomes. The conditional binomial test in such a matched-pair design is the exact McNemar test. However, in many clinical trials, one wants to establish equivalency between two treatments. We discuss how to use a conditional binomial test to establish equivalency between two treatments in a matched-pair design. Sample size and power determination for each conditional binomial test are calculated. Some statistical properties of the tests are analyzed through Monte Carlo simulation. PMID- 9855033 TI - A comparison of the random-effects pattern mixture model with last-observation carried-forward (LOCF) analysis in longitudinal clinical trials with dropouts. AB - The last-observation-carried-forward imputation method is commonly used for imputting data missing due to dropouts in longitudinal clinical trials. The method assumes that outcome remains constant at the last observed value after dropout, which is unlikely in many clinical trials. Recently, random-effects regression models have become popular for analysis of longitudinal clinical trial data with dropouts. However, inference obtained from random-effects regression models is valid when the missing-at-random dropout process is present. The random effects pattern-mixture model, on the other hand, provides an approach that is valid under more general missingness mechanisms. In this article we describe the use of random-effects pattern-mixture models under different patterns for dropouts. First, subjects are divided into groups depending on their missing-data patterns, and then model parameters are estimated for each pattern. Finally, overall estimates are obtained by averaging over the missing-data patterns and corresponding standard errors are obtained using the delta method. A typical longitudinal clinical trial data set is used to illustrate and compare the above methods of data analyses in the presence of missing data due to dropouts. PMID- 9855034 TI - A note on the small sample behavior of logistic regression in a bioassay setting. AB - The logistic regression procedure is a popular statistical method used when analyzing quantal dose-response data. However, logistic regression results based on a poorly designed experiment can be seriously compromised. Our results indicate that depending on the spacing of the doses, the number of doses, and the number of replications at each dose, the user can get very misleading results, including ineffective lack-of-fit tests and severely biased coefficient estimates along with biased estimates of response. In addition, variance formulas based on asymptotic theory may be completely inappropriate. Simulation results are used to support these statements. PMID- 9855035 TI - A note on the sample size determination in two-period repeated measurements crossover design with application to clinical trials. AB - The two-period repeated measurements crossover design is often used in clinical trials. In this article we give a formula for sample size determination for testing treatment effect in two-period repeated measurements crossover design by taking an analysis of variance approach to the repeated measurements analysis. A balanced situation is considered: two treatment sequences that have the same number of patients, and the time points of measurements on each subject within each treatment period equal in number. The formula reveals the relationship between the required number of patients for each treatment sequence, the number of repeated measurements within each treatment period, the detectable treatment difference with respect to a primary response variable, and the power for testing the treatment difference. A clinical trial example is given to illustrate the use of the formula. PMID- 9855037 TI - Effect of delayed observations on Bayesian decisions for binary data. AB - A Bayesian decision rule for early termination of an experimental study of binary responses is presented. This early termination occurs when the predictive probability of reversing the decision when utilizing the delayed observations is small. The proposed approach utilizes Bayesian inferential tools such as Bayes factors and predictive distributions. A simulation study is conducted to evaluate the performance of the proposed approach. Some guidelines are given to determine when the study should be terminated early and when the investigator should wait for delayed observations before making a conclusion. PMID- 9855036 TI - A power study of a sequential method of p-value adjustment for correlated continuous endpoints. AB - In the course of clinical (or preclinical) trial studies, it is a common practice to conduct a relatively large number of tests to extract the maximum level of information from the study. It has been known that as the number of tests (or endpoints) increases, the probability of falsely rejecting at least one hypothesis also increases. Single-step methods such as the Bonferroni, Sidak, or James approximation procedure have been used to adjust the p-values for each hypothesis. To reduce the conservatism (i.e., underestimating type I error) possessed by the aforementioned methods, Holm proposed a so-called "free-step down" procedure. This adjustment can be made even less conservative by incorporating the dependence structure of endpoints at each adjustment step of the procedure. That is done by sequentially applying James's approximation procedure for correlated endpoints at each step, referred to as the Free-James method. This article primarily compares the power of the Free-James method to the power of the Bonferroni and James single-step-down and the Holm free-step-down methods. Two definitions of power are considered: (a) the probability of correctly rejecting at least one hypothesis when it is true, and (b) the probability of correctly rejecting all hypotheses that are true. Monte Carlo simulations show that the Free-James method is as good as other methods under definition (a) and the most powerful under definition (b) for various sample sizes, numbers of endpoints, and correlations. PMID- 9855038 TI - Survival and projection analyses of the effect of radiation on beagle dogs. AB - The Cox proportional hazard model is used to identify important covariates related to survival for this beagle dog study. The Weibull and the Gompertz parametric probability models are used to fit the survival curves for 1680 beagle dogs given whole body Co-60 gamma radiation or sham irradiation and held for life span observation. Deaths from cancer and noncancer are the primary diagnoses. Of dogs dying from cancer, female dogs showed significantly greater cancer mortality (p < 0.0001) than did males, and irradiated dogs had significantly greater cancer mortality (p = 0.022) than did the unexposed dogs, by either the Weibull or the Cox model. However, there were no sex and exposure differences in mortality for dogs dying from noncancer causes. The fitted Weibull and Gompertz models have been successful in projecting the actual mortality experience in this experiment and could be used for similar life-span experiences. PMID- 9855039 TI - Multiple-objective optimal designs. AB - We present illustrative multiple-objective optimal designs for nonlinear models useful in biomedical studies. The aim is to demonstrate the added benefits of using a multiple-objective design over the traditional single-objective design. PMID- 9855040 TI - A global model to define the behavior of partial agonists (bell-shaped dose response inducers) in pharmacological evaluation of activity in the presence of the full agonist. AB - The dose-response models for full agonists and for a particular type of partial agonist can be described by sigmoidal curves and bell-shaped curves, respectively. The methods currently used to evaluate the interaction of a full agonist and a partial agonist require a large number of experimental units and base their analysis on nonlinear regression analysis, which may not be statistically appropriate. We propose an appropriate design and a global nonlinear model to evaluate such interactions. The new model allows us to estimate the interaction parameters and the parameters that characterize the individual partial agonist curve and the full agonist curve. PMID- 9855041 TI - Helicobacter pylori and somatostatin cells. AB - Tham et al. show that Helicobacter pylori infection lowers the density of immunoreactive somatostatin cells (D-cells) in the antral mucosa and elevates plasma gastrin concentrations. According to current hypothesis, the lack of inhibition by somatostatin allows excessive release of gastrin, which stimulates acid secretion and thus causes duodenal ulcers. The cytokine tumour necrosis factor-alpha which is released in H. pylori gastritis inhibits D-cells in culture and may be responsible. Why do not all infected persons get duodenal ulcers? Recent work shows that more aggressive strains of H. pylori have greater effects on somatostatin/gastrin physiology. Another variable is whether the infection causes corpusitis or not. Inflammation of the gastric corpus diminishes acid secretion, which greatly decreases the likelihood of duodenal ulcers but increases the risk of gastric cancer. Factors which promote corpusitis include diets with high salt content or lacking in antioxidant vitamins. Work in this area is elucidating how H. pylori causes different diseases. Hopefully this will allow us to predict and prevent its serious sequelae. PMID- 9855042 TI - Gastrin and colorectal cancer: where now? AB - Gastrin exerts a trophic influence on various regions of the gastrointestinal (GI) tract and this has led to an interest in its potential role in the growth of GI tumours. There is little evidence that elevated circulating levels of gastrin predispose to colonic tumours. However, the hormone can be detected within some colonic tumour tissues and a possible paracrine or autocrine role has been proposed. At present, evidence for such a role is conflicting, as is the evidence that colonic tumour cells possess receptors for the mature hormone. Colonic tumours have been found to contain much higher concentrations of incompletely processed gastrin precursors such as glycine extended gastrin and recent studies indicate that they may exert trophic effects mediated by specific receptors. Further studies of this are required. Whether specific hormone receptor antagonists will have a role in the clinical management of colonic tumours remains unclear. PMID- 9855043 TI - Effect of Helicobacter pylori eradication on antral somatostatin cell density in humans. AB - OBJECTIVE: As Helicobacter pylori infection is associated with an elevation in plasma gastrin with normal antral gastrin cell counts, an abnormality in antral somatostatin cells may be associated with the infection. We evaluated the effect of eradication of H. pylori on antral somatostatin cell density in the light of antral gastrin cell density and plasma gastrin levels. DESIGN: Prospective study. METHODS: Of 25 dyspeptic patients with H. pylori infection, nine had H. pylori successfully eradicated and the rest remained infected. Antral biopsies were immunostained for somatostatin cells and plasma gastrin measured before and 4 weeks after H. pylori eradication therapy. Ten other dyspeptic patients without H. pylori infection had their somatostatin cell density evaluated as controls. RESULTS: Somatostatin cell density in the patients without H. pylori infection at the outset was significantly higher than that in the patients with H. pylori infection at the outset (median 57 [18-83] vs. 37 [6-80] cells/mm) respectively (P <0.05). Somatostatin cell density increased after H. pylori eradication (before treatment, median 50 [15-72]; after treatment 71 [39-107] cells/mm) (P < 0.05) but was unchanged with persistent H. pylori infection. Plasma gastrin decreased after H. pylori eradication (before treatment, median 70 [45-100]; after treatment 30 [10-100] ng/l) (P < 0.05) but was unchanged with persistent H. pylori infection. CONCLUSIONS: Following eradication of H. pylori, there is an increase in somatostatin cell density with a fall in plasma gastrin. This supports the theory that H. pylori infection results in a decrease in somatostatin cell density and, as the latter is an inhibitor of gastrin cells, this results in an increased plasma gastrin. PMID- 9855044 TI - Endogenous hypergastrinaemia does not promote growth of colonic mucosa or of a transplanted colon adenocarcinoma in rats. AB - BACKGROUND/OBJECTIVE: Gastrin is trophic for the mucosa of the acid-producing part of the rat stomach, notably the histamine-producing ECL cells. Gastrin is said to stimulate growth also of colonic mucosa and colon cancer. The purpose of the present study was to examine whether endogenous hypergastrinaemia had trophic effects on normal colonic mucosa and transplanted colon adenocarcinoma in rats. METHODS: Rats were subjected to fundectomy (surgical removal of the acid producing part of the stomach) or treatments known to induce endogenous hypergastrinaemia. The treatments included refeeding after 48 h of food deprivation or administration of omeprazole (400 micromol/kg/day, orally). Other operations included colostomy and sham operation. A K12-cell line, originally established from a 1,2-dimethylhydrazine-induced colon adenocarcinoma, was used for transplantation. The rates of cell proliferation were determined in the oxyntic and colonic mucosa and in the tumour by measuring the proportion of the cells that accumulated bromodeoxyuridine in their nuclei, i.e. the labelling index (LI). The thickness of the oxyntic mucosa and the activity of histidine decarboxylase (HDC), the histamine-forming enzyme of the ECL cells, were measured. In addition, the thickness of the colonic mucosa and the weight and volume of the tumour were measured. RESULTS: Refeeding or treatment with a single dose of omeprazole in fasted rats raised the serum gastrin concentration and the LI and HDC activity in the oxyntic mucosa; refeeding but not omeprazole raised the LI in the colonic mucosa. In fed rats, hypergastrinaemia induced by fundectomy or treatment with omeprazole (for 10 days) failed to affect either the LI or the thickness of the mucosa of the proximal colon and the excluded distal colon of the colostomized rats. Fundectomy failed to stimulate the growth of the tumour transplants. CONCLUSION: Endogenous hypergastrinaemia did not induce trophic effects on rat colonic mucosa and did not promote growth of a transplanted colon adenocarcinoma in the rat. PMID- 9855045 TI - Lack of correlation between self-reported symptoms of dyspepsia and infection with Helicobacter pylori, in a general population sample. AB - OBJECTIVE: To investigate any correlation between infection with Helicobacter pylori (H. pylori) and overall symptoms of dyspepsia, in a general population sample. DESIGN: Analysis of test results and questionnaire replies from a population screening study involving subjects registered at a single general practice in Market Harborough, UK. METHODS: H. pylori status was established using a commercial enzyme-linked immunosorbent assay (ELISA), and frequent dyspepsia was assessed using a previously validated self-completion symptom questionnaire. RESULTS: Complete results for dyspepsia and H. pylori status were obtained and analysed for 1524 men and women aged 21-55 years at the start of the study. In those who attended for screening, the prevalence of dyspepsia was 39%, with a 15% prevalence of infection with H. pylori. No significant correlation was found between H. pylori status and frequent dyspepsia, upper abdominal pain or reflux-like symptoms. Adjustment for age, gender, smoking and alcohol consumption did not alter these findings. CONCLUSION: The analysis suggested that H. pylori infection does not play an important role in overall symptoms of non-ulcer dyspepsia in the community, nor is it important in protecting against acid reflux in patients without duodenal ulcer. PMID- 9855046 TI - Completeness of vagotomy, Helicobacter pylori colonization and recurrent ulcer 9 and 14 years after operation in duodenal ulcer patients. AB - OBJECTIVE: To clarify the relationship between the completeness of vagotomy and Helicobacter pylori colonization in the development of recurrent ulcer (RU) during a long-term follow-up period after the operation in duodenal ulcer (DU) patients. DESIGN: 122 consecutive vagotomized DU patients were studied twice on average 9 and 14 years after vagotomy. METHODS: The presence of RU and completeness of vagotomy were assessed simultaneously endoscopically and by endoscopic Congo Red test (ECRT). The positive ECRT showed incomplete vagotomy. The amount of H. pylori in the biopsy specimens of the gastric antrum and corpus mucosa was detected histologically by microscopic counting. RESULTS: The cumulative increase in RU occurred from 4% (5/122) at 9 years to 18% (22/122) at 14 years (P < 0.001) and the rate of ECRT positive cases rose from 52 to 71%, respectively (P < 0.01). All RU cases were ECRT positive. H. pylori colonization occurred in 92% of cases at 9 years and in 98% of cases at 14 years. Vagotomy increased H. pylori prevalence in the corpus mucosa and the rate of the high intensity grade of H. pylori in the antrum and corpus mucosa. CONCLUSION: The number of RU after vagotomy increases with time and is limited to patients with incomplete vagotomy. H. pylori colonization and the increased rate of its high intensity in the gastric mucosa after vagotomy may promote the development of RU only in incomplete vagotomy cases. PMID- 9855048 TI - Calcitonin gene-related peptide protects cultured rat gastric mucosal cells. AB - Capsaicin exerts its gastroprotective effect by stimulating primary afferent neurons, releasing calcitonin gene-related peptide (CGRP), which in turn increases gastric blood flow. In this work, the effects of capsaicin, rat alpha CGRP, and relative peptides hCGRP(8-37) and beta-hCGRP, and substance P on cultured gastric mucosal cells independent of neural and vascular mechanisms were studied. Damage was produced by indomethacin, ethanol or taurocholate 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue exclusion tests were used to assess viability of the cultured cells. Capsaicin administration alone did not injure gastric cells. However, capsaicin pretreatment potentiated the damaging effect of indomethacin and ethanol. In the sodium taurocholate model, capsaicin slightly protected the cells against injury. Alpha-rCGRP was protective against indomethacin, ethanol and taurocholate in a dose-dependent manner. hCGRP(8-37) and beta-hCGRP both dose-dependently prevented injury caused by indomethacin at concentrations about eight times higher than that of alpha-rCGRP, but substance P was ineffective in the three different damage models. A combination of alpha-CGRP and hCGRP(8-37) was also protective against indomethacin damage to a similar extent as use of either agent alone. The defence mechanism of capsaicin against gastric cell injury may in part be mediated by a direct effect of CGRP on gastric mucosal cells, in addition to effects dependent on neural and vascular mechanisms. hCGRP(8-37) has no antagonist effect against CGRP in this model, suggesting that CGRP receptors in this model may be different from those in other tissues. PMID- 9855047 TI - Does CagA contribute to ulcer pathogenesis in a developing country, such as Sudan? AB - BACKGROUND AND OBJECTIVE: In Western populations, peptic ulcer disease is closely associated with Helicobacter pylori (H. pylori) strains expressing the CagA antigen. In Africa the prevalence of H. pylori infection and peptic ulcer disease is high, although information regarding potential virulence factors is lacking. This study examines the prevalence of antibodies to CagA both in African patients with dyspepsia who are undergoing gastroscopy, and in asymptomatic healthy African volunteers. METHODS: Eighty two consecutive patients (median age 34 years, range 15-73 years), attending for gastroscopy were studied, of whom 78 (95.1%) were subsequently found to be Helicobacter positive. Three antral biopsies were obtained from each patient and 5 ml of blood was taken for determination of CagA seropositivity using western blot analysis. CagA seropositivity was also determined in 65 H. pylori positive healthy volunteers (median age 30 years, range 18-70 years), with no symptoms or previous history of gastroduodenal disease. RESULTS: Of the 78 H. pylori positive patients, CagA seropositivity was present in all 22 patients with active peptic ulcer disease (100%), in eight of nine patients with duodenitis (89%), in 15 of 19 patients with macroscopic gastritis (78.9%), and in 24 of 28 patients with a normal endoscopy (85.7%). On histological assessment, 46 patients had chronic active gastritis, 29 patients had gastritis with atrophy and three patients had intestinal metaplasia. CagA seropositivity rates were 84.7%, 93% and 100%, respectively, for these groups. In the 89 healthy volunteers studied, 57 of the 65 H. pylori positive subjects (87.7%) were seropositive for the CagA protein. CONCLUSIONS: As in Western countries, CagA seropositivity in this African population was closely related to endoscopic gastroduodenal disease, and to the presence of more advanced histological lesions in the antrum. However, there was also a high prevalence of CagA seropositivity in asymptomatic healthy individuals, suggesting that factors other than CagA predominate in ulcer pathogenesis in this population. PMID- 9855049 TI - CD4+ CD8+ and CD8alpha+ beta- T lymphocytes in human small intestinal lamina propria. AB - OBJECTIVE: To examine CD8 expression by T-lymphocyte subpopulations from disease free human lamina propria. METHODS: Single-cell suspensions were prepared from the epithelial layer and the lamina propria of small intestinal biopsies obtained endoscopically from disease-free patients. Monoclonal antibodies against CD3, CD4, CD8, CD56, CD8alphabeta, CD8alpha, TCR alphabeta and TCR gammadelta were used for dual and three-colour flow cytometric analysis. RESULTS: In addition to classical CD4+ and CD8+ T lymphocytes a substantial proportion of lamina propria T lymphocytes were CD4+ CD8+ or 'double positive' (mean 14%, range 4-26%). This population was significantly lower in the epithelial layer of the same patients (mean 7%, range 3-21%, P < 0.007). Three-colour flow cytometric analysis revealed that expression of the CD8 molecule on double positive T cells in the lamina propria was limited to the CD8alpha chain. Furthermore, of the CD8+ population, CD8+ T cells which only expressed the alpha chain were present in greater numbers in the lamina propria (mean 35%, range 14-54%) than in the epithelial layer (mean 18%, range 5-37%, P < 0.02). NK (CD56+) cells were not detected and few gammadeltaTCR+ T lymphocytes were detected in the lamina propria (mean 2%, range 0.5-6.6%) when compared with the epithelial layer (mean 8%, range 0.2-14%, P < 0.008). CONCLUSION: A significant population of CD4+ CD8alpha+ T lymphocytes which are CD8beta chain negative have been detected in the intestinal lamina propria. These cells form a more significant component of the lamina propria than the epithelial layer T-cell repertoire and may have a unique function in intestinal immunoregulation. PMID- 9855050 TI - Zinc, copper and immunological markers in the circulation of well nourished patients with ulcerative colitis. AB - OBJECTIVE: Few studies have been carried out on the trace element status in patients with ulcerative colitis (UC). Many trace elements are critical for the normal development and function of the immune system. This study was conducted in order to assess the serum levels of zinc and copper and the possible interrelation(s) between them and various immunological markers in the circulation of well nourished patients with UC. DESIGN/METHODS: The serum levels of zinc, copper, soluble interleukin-2 receptors (sIL-2Rs), interleukin-1beta (IL 1beta), interleukin-2 (IL-2), tumour necrosis factor-alpha (TNF-alpha), non-organ specific autoantibodies (RF, ANA, ANCA, anti-dsDNA and anticardiolipin), C3C and C4 components of the complement system and ceruloplasmin were determined in 75 well nourished patients with UC (32 patients with active and 43 with inactive disease). Thirty-three healthy individuals were also investigated. RESULTS: The mean concentrations (microg/dl) of zinc and copper were significantly higher (P < 0.0005 and P = 0.0001, respectively) either in active (202.3 +/- 115.2 and 141.7 +/- 31.4, respectively) or in inactive disease (204.5 +/- 170.3 and 137.4 +/- 24.5, respectively) compared with healthy controls (93.6 +/- 49.8 and 85 +/- 41.2, respectively). The levels of copper were positively correlated with the C3C (r = 0.41, P < 0.0005), C4 (r = 0.38, P < 0.001) and ceruloplasmin (r = 0.44, P < 0.0005), whereas zinc was correlated with C3C (r = 0.32, P = 0.0005) and ANA (P = 0.01). Autoantibodies of at least one specificity (AUBS) were found in 77.3% of the patients. The mean levels (U/ml) of sIL-2Rs were significantly higher (P = 0.0001) in active disease (604.3 +/- 213.0) than in inactive UC (411.5 +/- 165.1) and in patients with ANA (P < 0.05), ANCA (P = 0.01) or AUBS (P < 0.05). The sIL 2Rs were correlated with the C4 (r = 0.40, P < 0.005) and the ESR (r = 0.43, P = 0.0001). CONCLUSION: These findings indicate that even in well nourished patients with UC, high serum levels of copper and zinc are present. The latter alterations of zinc and copper are correlated with haematological parameters of relapse of the disease or with acute phase proteins suggesting a relationship with the inflammatory process of UC. Further studies on the colonic tissue will address the role of zinc and copper in the inflammatory and immune reactions observed in this disease process. PMID- 9855051 TI - Lactulose and neomycin attenuate leukocyte-endothelial cell adhesion in an animal model of inflammatory bowel disease. AB - OBJECTIVE: The role of intestinal flora in the pathogenesis of inflammatory bowel disease is under discussion. The objective of this study was to assess the effect of lactulose (Lac) and neomycin (Neo), both of which are used clinically for changing or reducing intestinal flora, on leukocyte-endothelial cell interaction in an indomethacin (Indo)-induced long-lasting ileitis in Sprague-Dawley rats. METHODS: Two doses of Indo (7.5 mg/kg, s.c.) were given 24 h apart. Animals were fed with standard rat chow for 10 days until 12 h prior to the experiment. Solutions of Lac (1.0 mg/kg b.w.) or Neo (0.1 g/kg) were gavaged daily for 10 days between Indo administration and the experiment. Ten mesenteric venules (30 microm diameter) per animal (n = 5 per group) were observed using intravital microscopy, and the following parameters were monitored: number of adherent and emigrated leukocytes, leukocyte rolling velocity, erythrocyte velocity, venular blood flow, and shear rate. Macroscopically visible injury was scored 0 to 5, and faecal pH was measured in the distal 20 cm of ileum. RESULTS: Ten days after Indo treatment leukocyte adherence and emigration were increased (2.2-fold and 3.3 fold vs. control, respectively) while leukocyte rolling velocity and venular wall shear rate were reduced (both parameters to 81% of control). Lac and Neo attenuated microcirculatory parameters to a similar extent while macroscopic damage was prevented only by Neo but not by Lac. The effects of both substances were accompanied by a normalization of faecal pH to control level which had been lowered by Indo. CONCLUSION: The Indo-induced increase in leukocyte-endothelial cell interaction is blunted by Lac and Neo. At the same time the Indo-induced lowering of faecal pH is normalized by both substances. The reduction of macroscopic injury by Neo but not by Lac in the chronic phase of Indo inflammation might be due to additional effects on other inflammatory cells such as macrophages. PMID- 9855052 TI - Diagnostic value of CA 72-4 and carcinoembryonic antigen determination in the fluid of pancreatic cystic lesions. AB - OBJECTIVE: Mucinous cystic tumours of the pancreas need to be distinguished from other cystic lesions because of their malignant potential. The aim of this study was to assess prospectively the reliability of CA 72-4 and carcinoembryonic antigen analysis in the fluid of cystic lesions of the pancreas obtained by fine needle aspiration for pathological diagnosis. METHODS: CA 72-4 and carcinoembryonic antigen were measured in cyst fluid obtained preoperatively by fine-needle aspiration. The 91 lesions consisted of 16 serous cystadenomas, 16 mucinous cystadenomas, 14 cystadenocarcinomas and 45 pancreatic pseudocysts complicating well documented chronic pancreatitis. RESULTS: A CA 72-4 level of >40 U/ml had a 63% sensitivity and 98% specificity for distinguishing mucinous cystadenomas and cystadenocarcinomas from serous cystadenomas and pseudocysts. A carcinoembryonic antigen level of >400 ng/ml had a 57% sensitivity and a 100% specificity for distinguishing mucinous tumours and cystadenocarcinomas from pseudocysts. A carcinoembryonic antigen level of <4 ng/ml had a 100% sensitivity and a 93% specificity for distinguishing serous cystadenomas from mucinous cystadenomas, cystadenocarcinomas and pseudocysts. CONCLUSION: Combined measurement of CA 72-4 and carcinoembryonic antigen may be used to distinguish accurately mucinous cystadenomas and cystadenocarcinomas from serous cystadenomas and pseudocysts. PMID- 9855053 TI - The development of hepatic granulomas following interferon-alpha2b therapy for chronic hepatitis C infection. AB - Hepatic granulomas have been detected in cases of chronic hepatitis C virus (CHCV) infection. Here we report upon three selected cases of CHCV infection who were treated with IFN alpha, in whom hepatic granulomas were absent on initial liver biopsies, but in whom granulomas developed following IFN alpha therapy. In one case, complete regression of these granulomas was noted 17 months following discontinuation of treatment, more strongly implicating IFN alpha in the aetiopathogenesis of such granulomas in this case. These findings suggest that IFN alpha may have a biological role in the pathogenesis of granulomatous liver disease in these patients. All three cases were poor/non-responders to IFN alpha, suggesting that the development of hepatic granulomas during IFN alpha therapy may be associated with a poor response to treatment. PMID- 9855054 TI - Fatal pneumococcal septicaemia in a coeliac patient. AB - A 78-year-old lady initially presented with painful hips, low back pain, lethargy and weight loss. She had a past history of osteomalacia. Investigations revealed evidence of malabsorption and jejunal biopsy revealed sub-total villous atrophy in keeping with coeliac disease. Peripheral blood film was within normal limits. She responded well clinically to a gluten-free diet and calcium and vitamin D supplementation. Four years after the initial diagnosis she presented acutely with vomiting, pleuritic chest pain, pyrexia and bronchospasm. Blood cultures confirmed the presence of Streptococcus pneumoniae and she was treated appropriately with ampicillin. Despite this she died shortly after admission. It is recognized that blood film examination alone cannot exclude hyposplenism complicating coeliac disease and it is presumed that this was the reason for the development of fatal pneumococcal septicaemia in this patient. Prophylactic vaccination may be appropriate in hyposplenism secondary to coeliac disease. PMID- 9855055 TI - IgM antibody against measles virus in patients with inflammatory bowel disease: a marker of virus related disease? PMID- 9855056 TI - Helicobacter pylori: a true carcinogen? AB - Is Helicobacter pylori a true carcinogen? Most carcinogens are physical, chemical or viral agents which give rise to the development of neoplasia by inducing alterations in cellular DNA. Evidence for the carcinogenic potency of such agents is usually based on dose-response curves and animal models and there is often a direct association with (epi-) genetic events. Despite the absence of such data, H. pylori has been designated as a definite cause of human cancer. This designation is largely based on epidemiological evidence. H. pylori is a carcinogen in the sense that infection with this organism induces a persistently inflamed gastric mucosa which is associated with an increased proliferative state and an increased gastric cancer risk. As such, 1-2% of the infected subjects are estimated to develop cancer, with an incidence probably close to nine times higher than that among non-infected subjects. There is a need for additional mechanistic knowledge on this association between chronic epithelial inflammation and carcinogenesis. An additional important question is whether H. pylori eradication may contribute to gastric cancer prevention. As progressive mucosal abnormalities such as atrophy and metaplasia do not regress after such intervention, the major benefit in terms of cancer prevention is likely to be to infected subjects who have not yet developed permanent gastric mucosal damage. This is in agreement with data suggesting that the role of H. pylori may be confined to the initial stages of carcinogenesis. PMID- 9855057 TI - Quality of life assessment in reflux disease. AB - The recognition of heartburn and acid regurgitation as manifestations of gastro oesophageal reflux disease (GORD), and the suppression of these symptoms with therapy, represent an over-simplistic approach to the reflux patient. For GORD, as for other gastrointestinal disorders, it is evident that many patients suffer a greater impairment of well-being than has been appreciated hitherto, and that this impairment can be quantified using modern quality of life measuring techniques. Successful treatment normalizes quality of life in a manner which is substantially, but not wholly, predictable from symptom responses or the healing of oesophagitis. These observations raise the possibility that physicians' therapeutic intentions do not always match the patients' wishes. The appraisal of health status, or quality of life (QOL), is increasingly important in defining the implications of disease and for assessing the outcome of therapy. Hitherto, QOL evaluations have seemed relevant to circumstances of major physical disability and for patients receiving potentially unpleasant treatments, such as cancer chemotherapy, but it is now evident that QOL evaluation has much wider applicability, including relevance to common gastrointestinal disorders such as GORD. PMID- 9855058 TI - Is appendectomy a causative factor in ulcerative colitis? AB - There are strong indicators that the aetiology of inflammatory bowel disease should be regarded as multifactorial, involving an interaction between genetic and environmental factors which give rise to an inadequate immunological response. During the past decade at least seven case-control studies have shown an inverse association between appendectomy and ulcerative colitis. Conclusions have been that either ulcerative colitis protects against appendicitis, or appendectomy protects against ulcerative colitis. The immunological function of the appendix is not well known, but experimental studies suggest that the appendix is possibly an important site for priming of the cells involved in the development of inflammatory bowel disease. Experimental and prospective cohort studies are needed to provide more insight in a possible relation between ulcerative colitis and the appendix. PMID- 9855059 TI - The effect of treating reflux oesophagitis with omeprazole on quality of life. AB - BACKGROUND/AIMS: Despite the commonplace nature of heartburn and reflux oesophagitis, little is known of their impact on patients' quality of life. The aim of this study was to assess quality of life in oesophagitis patients before and after medical therapy and compare the results with a sample of the general population. METHODS: Consecutive attenders with frequent heartburn and grade II III oesophagitis on endoscopy were recruited from one of two centres and treated with omeprazole 20 mg BD for 8-14 weeks. A symptomatic questionnaire, including the Short Form-36 (SF-36) quality of life questionnaire, was completed before and at the end of treatment. Actual quality of life scores were compared with 'expected' scores derived from a sample (n = 3015) of the Northern Ireland population. RESULTS: Seventy two (83%) of the 87 patients recruited were healed after 14 weeks therapy and 77 completed the SF-36 before and after therapy. Three quality of life parameters (bodily pain, vitality and social function) were significantly lower before treatment than the 'expected' scores. Seven of the quality of life parameters measured by the SF-36 showed a significant improvement after treatment and the eighth (mental health) just failed to achieve a significant improvement (P = 0.06). Comparison of the improvements in SF-36 scores for those who were healed with scores for those who were not healed showed no significant difference. CONCLUSIONS: Patients with grade II-III oesophagitis and frequent heartburn have lower quality of life scores for some parameters than would be expected in the general population. Treatment of the oesophagitis with omeprazole 20 mg BD causes a significant improvement not just in reflux symptoms but in several physical and mental aspects of quality of life regardless of whether or not the oesophagitis is healed. PMID- 9855060 TI - The prevalence of Helicobacter pylori gastritis in patients with reflux oesophagitis: a case-control study. AB - OBJECTIVES: To investigate the prevalence of Helicobacter pylori in patients with erosive oesophagitis in comparison with asymptomatic subjects. DESIGN: Hospital based case-control study. METHODS: Two hundred and nineteen consecutive patients with erosive oesophagitis diagnosed over 2 years were studied. Patients with secondary gastro-oesophageal reflux disease, prior H. pylori eradication therapy, PPI or antibiotics were excluded and two case groups were formed: (1) patients with oesophagitis alone (OEA), and (2) patients with oesophagitis and concomitant peptic ulcer disease (OE&PUD). H. pylori was assessed by antral and corpus mucosa histology and the rapid urease test. Age- and sex-matched controls, who underwent voluntary screening for H. pylori by the 13C-urea breath test, had neither symptoms nor a history of upper gastrointestinal disease. RESULTS: The 130 patients in the OEA group had a prevalence of H. pylori of 38.5% compared with 75.6% in the 41 patients in the OE&PUD group and 88.5% in a subgroup of the latter with duodenal ulcer (26 patients). Infected and non-infected cases showed no significant differences in terms of grade of oesophagitis and incidence of Barrett's oesophagus. The prevalence of H. pylori infection in OEA was similar to that found in matched controls. In OE&PUD the relative risk of H. pylori infection was 3.6-fold higher than in controls [P = 0.0069, 95% confidence interval (CI), 1.401-9.195]. CONCLUSION: The prevalence of H. pylori in oesophagitis without PUD is similar to that of the asymptomatic population with no history of upper gastrointestinal disease. When oesophagitis and PUD occur together, the infection rate is within the range found in PUD alone. PMID- 9855061 TI - Helicobacter pylori infection in patients affected by insulin-dependent diabetes mellitus. AB - BACKGROUND/AIM: Insulin-dependent diabetes mellitus (IDDM) is the result of an autoimmune destruction of pancreatic Langerhans beta cells. Helicobacter pylori infection, the most common cause of gastritis and peptic ulcer, has been associated with some autoimmune diseases. The aim of this study was to evaluate the prevalence of H. pylori infection and gastrointestinal (GI) symptoms in patients with IDDM. METHODS: 116 patients (50 males and 66 females, mean age: 35 +/- 11 years) affected by IDDM were evaluated; 50 healthy subjects matched for age and sex were utilized as a control group. H. pylori infection was determined by a [13C]urea breath test. IDDM duration, daily dosage of insulin and prevalence of GI symptoms (bloating, pyrosis, epigastric pain, belching, halitosis, nausea) were assessed. RESULTS: The prevalence of H. pylori infection was similar in IDDM patients and in controls (37 vs 34%, respectively). In both groups, the mean age was higher in infected subjects. The mean IDDM duration was significantly greater in infected patients (19 +/- 12 vs 13 +/- 10 years, P < 0.006). Prevalence of infection was directly related to the duration of IDDM (<1 year, 23%; 1-3 years, 32%; >3 years, 40%). The daily dosage of insulin did not differ between infected and non-infected subjects. H. pylori contaminated patients showed a significantly higher prevalence of bloating, pyrosis and epigastric pain when compared with non infected subjects; conversely, prevalence of belching, halitosis and nausea did not differ between groups. CONCLUSION: The prevalence of H. pylori infection is high in patients affected by IDDM; however, it does not differ from the infection rate observed in a control group. The rate of infection increases with IDDM duration. Bloating, pyrosis and epigastric pain appear to define infection status better. PMID- 9855062 TI - Topographic distribution of Helicobacter pylori in the resected stomach. AB - OBJECTIVES: The aim of this study was to elucidate the prevalence of Helicobacter pylori and its distribution in order to clarify the frequency of H. pylori infection and the most appropriate site of endoscopic biopsy for studies of H. pylori infection associated with different gastric diseases. DESIGNS AND METHODS: Swiss role mucosal strips from 275 resected stomachs, which included the greater curvature, anterior wall and lesser curvature of the antrum, incisura and corpus, were stained with haematoxylin-eosin and H. pylori antibody. RESULTS: The prevalence of H. pylori infection was 97% in duodenal ulcers, 98% in gastric ulcers, 98% in intestinal-type carcinomas and 99% in diffuse-type carcinomas. H. pylori was present at a rate of 100% in any site in cases of duodenal ulcer, but was diffusely distributed in the antrum and patchily distributed in the corpus. The detection rate of H. pylori was 50-100% in gastric ulcers, 30-100% in intestinal-type adenocarcinomas and 63-100% in diffuse-type adenocarcinomas depending on the site of the stomach examined. CONCLUSIONS: The prevalence of H. pylori infection was very high in peptic ulcers of the duodenum and stomach and gastric carcinomas of Japanese patients. Biopsy specimens for evaluation of H. pylori infection should be taken routinely from both the greater curvature of the antrum and corpus. Immunohistochemical staining should be used to assay for H. pylori when few organisms are present or eradication therapy has been used. PMID- 9855063 TI - 5T4 oncofetal antigen in gastric carcinoma and its clinical significance. AB - OBJECTIVE: To evaluate the role of 5T4 antigen in gastric cancer progression and prognosis. DESIGN: A prospective study of 5T4 antigen expression in primary, secondary and recurrent gastric carcinoma, the relationship to selected prognostic parameters and the course of disease. PATIENTS: Eighty six patients operated on for gastric cancer. TISSUE: One hundred and twenty two gastric tumours were studied, including 86 primary carcinomas, 32 coexisting lymph node metastases and four recurrent carcinomas. METHODS: Immunohistochemistry using 5T4 monoclonal antibody on frozen sections. RESULTS: The 5T4 antigen was detected in 41% of primary gastric tumours including early gastric cancer. A strong relationship was found between 5T4 positivity and tumour histology. Thus, 52% of gastric carcinomas of intestinal type expressed 5T4 antigen compared with 28% of the diffuse type (P = 0.028). Among 16 sets of primary gastric carcinomas and regional lymph node metastases, coordinate 5T4 expression was seen in 14 cases; the other two showed acquisition of positivity on metastatic tumour cells (carcinomas of diffuse type). 5T4 antigen was detected more frequently in carcinomas with p53 accumulation compared with those with undetectable p53 levels (P = 0.015). The presence of 5T4 in cancer cells was correlated with poor short term prognosis (24% vs 49% of 2 year survival for 5T4 positive and negative tumours respectively, P = 0.024). The effect on survival was evident in the p53 negative group, with patients 5T4 positive showing worse survival (28% vs 60% in 2 years). CONCLUSIONS: Our results suggest that the assessment of 5T4 expression in gastric carcinoma can be helpful in identifying patients with poor short-term prognosis. PMID- 9855064 TI - The impact of chronic hepatitis C virus infection on HIV disease and progression in intravenous drug users. AB - OBJECTIVE: It is unclear whether co-infection with hepatitis C virus (HCV) can influence HIV related morbidity or mortality, either by accelerating HIV-related disease progression, or by contributing to end stage liver disease. The aim of this study was to examine the effect of HCV infection on the severity and progression of HIV disease in a cohort of Edinburgh intravenous drug users (IDUs). METHODS: In 240 (47%) out of 508 patients in the Edinburgh IDU cohort both HIV seroconversion dates and anti-HCV serology were available. Demographic variables and HIV-related progression between anti-HCV positive and anti-HCV negative groups were compared. Parameters assessed included clinical endpoints (time of development of significant symptoms attributable to HIV (CDC stage IV), time of development of AIDS, and time of death) and immunological endpoints (time of CD4+ counts dropping below 200/mm3, 100/mm3 and 50/mm3). RESULTS: Two hundred and two out of 240 patients (84%) had positive anti-HCV serology. There was no significant difference in the frequency of clinical and immunological endpoints between the anti-HCV positive and negative groups. Progression analysis from HIV seroconversion to HIV related clinical endpoints indicated that anti-HCV serology was not a significant factor influencing the rate of HIV progression (relative risks (RR) for anti-HCV positive group: seroconversion to CDC IV, 1.01; seroconversion to AIDS, 1.05; seroconversion to death, 0.90). Likewise, HCV serostatus did not significantly affect progression to immunological endpoints (RR for anti-HCV positive group: seroconversion to CD4+ < 200/mm3, 1.04; seroconversion to CD4+ < 100/mm3, 1.13; seroconversion to CD4+ < 50/mm3, 0.97. Overall mortality from end stage liver failure was 4% in HCV-seropositive patients without AIDS. This suggests that HCV has had a clinically (though not statistically) significant impact on overall survival in this cohort. CONCLUSIONS: This study demonstrates that HCV co-infection does not influence the rate of progression to either clinical or immunological endpoints in our population of HIV-infected drug users. Further data are required to assess the effect of HIV on thge progression of HCV-related liver disease. PMID- 9855065 TI - Effects of interferon-alpha on cytochrome P-450 isoforms 1A2 and 3A activities in patients with chronic hepatitis C. AB - BACKGROUND/AIM: The risk of adverse drug interactions with interferon-alpha has been poorly assessed. The aim of our study was to establish whether administration of interferon-alpha at therapeutic doses in patients with chronic hepatitis C may have significant inhibitory effects on other drug metabolism. The study was focused on cytochromes P-450 1A2 and 3A, two major isoforms involved in the metabolism of numerous substrates. METHODS: Eighteen patients with chronic active hepatitis C requiring an interferon-alpha treatment were studied. Cytochrome P-450 1A2 activity was determined on the basis of an in vivo caffeine metabolism study. Cytochrome P-450 3A activity was determined according to in vivo cortisol metabolism into 6-beta-hydroxycortisol. Both activities were determined 1 month before, at initiation and 1 month after interferon-alpha therapy (3 x 10(6) units, three times a week). RESULTS: There were no significant differences in the caffeine index (CYP 1A2) and in the 6-beta hydroxycortisol/free cortisol urinary ratio (CYP 3A) before and after alpha interferon treatment CONCLUSION: Chronic administration of interferon-alpha at therapeutic doses does not change in vivo cytochrome P-450 1A2 and 3A activities. These results support the suggestion that drugs metabolized by these isoenzymes may be used together with interferon-alpha in patients with chronic hepatitis C without significant risks of drug interactions. PMID- 9855066 TI - The acute phase protein alpha-1-antitrypsin inhibits transferrin uptake in PLC/PRF/5 cells and increases release of hepatitis B virus surface antigen and alpha-fetoprotein. AB - BACKGROUND/AIMS: The present study was designed to investigate whether the acute phase protein alpha-1-antitrypsin (alpha1-AT), which has an inhibitory effect on transferrin (tf) receptor-mediated iron uptake in K562 and THP1 cells, has a similar effect in PLC/PRF/5 cells. This hepatic cell line is of specific interest because it is infected with hepatitis B virus (HBV). Therefore, we addressed the additional question whether alpha1-AT has any effect on cellular protein synthesis and replication of HBV in PLC/PRF/5 cells. METHODS: Cells were incubated with various concentrations of alpha1-AT, dexamethasone, IL-6 and desferrioxamine. HBs-AG, alpha-fetoprotein and albumin concentrations in culture media were measured using commercially available methods. For equilibrium inhibition binding experiments, cells were incubated with 85-182 pmol/l [125I]tf. To study the potential effect of alpha1-AT on DNA synthesis we measured the incorporation of [3H]thymidine into DNA. RESULTS: In equilibrium saturation binding experiments, [125I]tf bound to PLC/PRF/5 cells with K(D) 17.45 +/- 4.57 nM and a maximum density of binding sites of 267,285 +/- 39,915 sites/cell. In inhibition studies alpha1-AT demonstrated an apparently monophasic inhibition of [125I]tf to its receptor. At concentrations > 30 micromol/l alpha1-AT inhibited the growth of PLC/PRF/5 cells up to approximately 50%. The inhibitory effect of alpha1-AT on DNA synthesis was not as potent as that on growth. At the highest concentration of 100 micromol/l, alpha1-AT produced a 35% maximum inhibition of [3H]thymidine incorporation. Incubating PLC/PRF/5 cells with corticosteroids enhanced HBs-AG release significantly. Interestingly, alpha1-AT showed the same pattern of effects on cell metabolism and HBs-AG release as the corticosteroids. When we incubated the cells with 50 micromol/l alpha1-AT, alpha-fetoprotein production increased significantly and HBs-AG release almost doubled. CONCLUSION: We have to assume that there is a specific mechanism inducing HBs-AG release by alpha1-AT, as has been shown to be the case with steroids. PMID- 9855067 TI - Effect of inhibition of ornithine decarboxylase activity in a model of acute hepatocellular necrosis. AB - OBJECTIVE: The effect of blockade of the enzyme ornithine decarboxylase by difluoromethylornithine (DFMO) on hepatocellular necrosis and survival in rats treated with thioacetamide (TAA) was investigated. DESIGN: In one experiment, the effect of DFMO on survival of rats with TAA-induced acute hepatocellular necrosis was determined. In another experiment, blood and liver specimens were obtained from DFMO or saline-treated rats 24 h after the administration of TAA for determinations of serum alanine aminotransferase (ALT) and liver content of polyamines and microsomal cytochrome P-450 and for assessment of hepatic histology. METHODS: Liver polyamines were determined by reversed-phase HPLC and microsomal cytochrome P-450 content by dithionite-difference spectroscopy of CO treated homogenates. The severity of hepatocellular necrosis was scored blindly. RESULTS: TAA-treated rats that received DFMO survived longer than saline-treated controls (P < 0.01). Serum ALT and liver putrescine concentrations were lower and the histological severity of acute hepatocellular necrosis was less in DFMO treated rats with TAA-induced hepatocellular necrosis than in saline-treated controls (P < 0.05, P < 0.01 and P < 0.05, respectively). Total cytochrome P-450 levels were similar in DFMO and saline-treated rats with TAA-induced hepatocellular necrosis. CONCLUSIONS: DFMO increases survival in TAA-induced fulminant hepatic failure by decreasing the severity of acute hepatocellular necrosis. The beneficial effects of DFMO do not appear to be mediated by its effects on polyamine metabolism, but may be attributable to an effect of DFMO on thioacetamide metabolism or on an alternative pathway of ornithine metabolism. PMID- 9855068 TI - Somatostatin for acute severe bleeding from portal hypertensive gastropathy. AB - OBJECTIVES: To investigate the effect of somatostatin in acute severe bleeding from portal hypertensive gastropathy in 26 cirrhotic patients. METHODS: All patients with signs of acute gastrointestinal bleeding and an upper GI endoscopy (during the first 24 h) indicating overt bleeding from portal hypertensive gastropathy were included in the study. Somatostatin (or the synthetic tetradecapeptide, octreotide) was administered in all cases. Eleven patients received somatostatin and 15 patients received octreotide. An initial injection of 250 microg bolus somatostatin was followed by a continuous infusion of 250 microg/h for 3 days (100 microg and 50 microg/h for octreotide). RESULTS: Somatostatin arrested bleeding in all 26 patients and in 23 there was no hospital relapse. In the remaining three patients the bleeding recurred each time somatostatin infusion was discontinued and arrested again on reinstitution of treatment. In two there was a control of haemorrhage, while the third required a total gastrectomy after repeated episodes. The rebleeding rate in our study is much lower compared to untreated patients of other series. There were no differences between the somatostatin and octreotide group. There were no significant side effects. Gastroscopy at the end of the therapy showed improvement of the endoscopic appearance. CONCLUSIONS: This open study suggests that somatostatin is a safe and effective treatment of acute severe bleeding from portal hypertensive gastropathy. PMID- 9855069 TI - Outcome of ulcerative colitis after treatment with transdermal nicotine. AB - OBJECTIVE: Transdermal nicotine appears to be of benefit in the short-term treatment of patients with ulcerative colitis. The aim of this study was to determine its long-term effects. DESIGN: A randomized, comparative study. PATIENTS AND METHODS: Patients with mild to moderate clinical relapses of left sided ulcerative colitis during maintenance treatment with mesalamine 1 g b.i.d. were allocated to an additional treatment with either transdermal nicotine or prednisone for 5 weeks. The first consecutive 15 patients per group, with clinical and endoscopic signs of remission, were followed up for 6 months, while continuing mesalamine maintenance treatment. RESULTS: Relapses of active colitis were observed in 20% of patients formerly treated with nicotine and in 60% of patients in the prednisone group (P = 0.027). Relapses occurred earlier in the latter group. CONCLUSION: Our results confirm that nicotine is useful in cases of ulcerative colitis with mild or moderate activity and suggest that remissions induced by nicotine may last longer than those obtained with oral corticosteroids. PMID- 9855070 TI - Colonic endocrine cells in patients with carcinoma of the colon. AB - OBJECTIVE: To investigate the colonic endocrine cells in patients with colon carcinoma in order to establish a possible abnormality. PATIENTS: Twelve patients with colon adenocarcinoma (eight women and four men; mean age 69 years; range 52 88 years) were studied. As controls, macroscopically and histologically normal tissues from the colon of 12 patients (eight women and four men; mean age 66 years; range 34-86 years) were examined. These patients suffered from polyps, prolapsus, chronic diverticulitis, volvulus or haemorrhoids. METHODS: Macroscopically and histologically normal tissues from the colon of the patients, about 10 cm from the tumour, and of the controls were examined. Endocrine cells were immunostained with the avidin-biotin-complex method, and were quantified by computer image analysis using an automatic standard sequence analysis operation. Three parameters were used: (1) the number of endocrine cells per mm3 of epithelial cells; (2) the cell secretory index (CSI); and (3) the nuclear volume. RESULTS: The numbers of somatostatin- and serotonin-immunoreactive cells were significantly reduced in patients with colon carcinoma (P = 0.03 and 0.009, respectively). There was no difference between patients and controls regarding the numbers of PYY-, PP-, and enteroglucagon-immunoreactive cells. The CSI of somatostatin- and serotonin-immunoreactive cells were significantly decreased. There was no difference in the CSI between the patients and controls regarding PYY-, PP- or enteroglucagon-immunoreactive cells. The nuclear volumes of PYY- and enteroglucagon-immunoreactive cells increased significantly in the patients. The nuclear volume of PP-, somatostatin- and serotonin cells did not differ from those of the controls. CONCLUSION: The present results support the assumption that a disturbance in the colonic neuroendocrine system occurs in patients with colon carcinoma, which might affect the development of the tumour. The decrease in the number and CSI of somatostatin cells may account for the decrease of the colonic content of this peptide observed previously in these patients. The decrease in the number and CSI of serotonin-immunoreactive cells in patients with colon adenocarcinoma might be a method by which the body defends itself against this mitogenic substance. PMID- 9855071 TI - Atrophy of the coeliac mucosa. AB - Details of 12 patients suffering from atrophy of the coeliac mucosa (ACM) are presented. These patients failed to respond to the elimination of gluten from their diets. Their jejunal biopsies were examined in detail with particular attention given to Paneth cell numbers, crypt length, mitotic rate and mucosal thickness. These biopsies were compared with the biopsies of responsive coeliac patients. The ACM patients were found to have a statistically significant reduction in Paneth cell numbers (P < 0.0005), reduced crypt length (P < 0.0125), reduced mitotic rate (P < 0.0005) and a thinner mucosa (P < 0.0005), when compared with the responsive coeliac group. In addition the ACM group had a lower albumin level compared with the responsive group (P < 0.0005). Most of the ACM patients responded when treated with oral corticosteroids. It is considered that the above features are markers of a non-responsive group of coeliac patients and thus might be useful for identifying those patients who may benefit from early treatment with oral corticosteroids if gluten withdrawal is not rapidly effective. PMID- 9855072 TI - Endoscopic treatment of wirsungo-cysto-pleural fistula. AB - A case is reported of a female with chronic alcoholic calcifying pancreatitis who presented with a wirsungo-cysto-pleural fistula. Endoscopic retrograde pancreatography demonstrated the fistulous tract and a naso-pancreatic drain was inserted. Subsequently, this drain was replaced by a pancreatic endoprosthesis. This endoscopic therapy led to full resolution of the fistula. We suggest that endoscopic intervention is the first-line treatment for this condition and that surgical intervention should be reserved as a second-line treatment. PMID- 9855073 TI - Pathogenesis of primary biliary cirrhosis: CD95-induced apoptosis at last? AB - Apoptosis is a basic mechanism involved both in maintaining tissue homeostasis by elimination of senescent or potentially harmful cells and in the regulation of immune responses. If not properly regulated, however, it may lead to serious tissue damage. CD95(Fas/APO-1) is a surface receptor that mediates apoptosis upon oligomerization by its ligand, CD95L. The CD95/CD95L-system is one of the major effectors of cytotoxicity in inflammation with implications for both the prevention and the pathogenesis of autoimmune diseases. In primary biliary cirrhosis (PBC), an autoimmune disease, apoptosis has been repeatedly suspected to be the mechanism leading to progressive destruction of bile ducts. The role of apoptosis and its possible molecular inducers in PBC are discussed. PMID- 9855074 TI - Endoscopic ultrasound: a new look at achalasia? AB - Achalasia is a relatively infrequent disorder of oesophageal motility, and is most often easily diagnosed by stationary manometry showing absence of peristalsis and incomplete lower oesophageal sphincter (LOS) relaxation. In rare cases manometric findings may be inconclusive or a malignant underlying disease is suspected. In such cases direct visualization of the LOS by endoscopic ultrasound (EUS) could contribute to securing the diagnosis and possibly lead to earlier diagnosis and treatment. With lower frequency EUS, increased thickness of the inner circular layer of the muscularis propria may be difficult to detect. Available data suggest that high frequency (20 MHz) and 3-D EUS has a potential to reliably discriminate between achalasia patients and normal subjects, and thereby be of diagnostic value. PMID- 9855075 TI - Gastro-oesophageal reflux in children. AB - Disordered motility of the gastrointestinal tract in children is usually manifest as gastro-oesophageal reflux disease. Reflux is common in infants but usually resolves spontaneously with time. If severe enough to merit investigation, 24 hour ambulant oesophageal pH monitoring is the most widely used investigation, though it does have disadvantages. Upper gastrointestinal endoscopy and biopsy may be required to diagnose oesophagitis. Treatment follows a step-wise approach, progressing from positioning and feed-thickening in infants, through to pharmacological management with prokinetic agents and/or acid-suppression, and surgery for a very small proportion of children whose symptoms do not respond to optimal medical therapy. PMID- 9855076 TI - A discussion of how terlipressin limits mortality in cases of bleeding oesophageal varices. AB - Bleeding oesophageal varices (BOV) are a potentially life-threatening complication of portal hypertension. While endoscopic sclerotherapy and banding ligation are often employed in an attempt to arrest bleeding, the use of vasoactive pharmacological agents to control haemorrhage has a number of advantages. While many of the available vasoactive agents control acute bleeding and may exert a beneficial influence over hepatic haemodynamics, terlipressin (triglycyl lysine-vasopressin) is the only agent that has been shown actually to decrease mortality in cases of BOV. It is hypothesized that this increase in survival rate is due to the apparently unique multifactorial influence of terlipressin over variceal haemostasis and blood flow, hepatic and gastric haemodynamics and renal function, combined with the likelihood of only minimal adverse events. PMID- 9855077 TI - Bile duct cells in primary biliary cirrhosis are 'primed' for apoptosis. AB - OBJECTIVE: Primary biliary cirrhosis (PBC) is characterized by progressive, immune-mediated destruction of bile ducts (<75 microm diameter) and secondary changes related to cholestasis which may involve apoptosis. In this study we sought to examine the protein expression of genes involved in apoptosis in biliary epithelium of PBC cases. DESIGN: In order to investigate the susceptibility of biliary epithelial cells to apoptosis and their ability to proliferate, we examined the expression of a number of apoptosis related proteins in early and late stage PBC and histologically normal liver control tissue using immunohistochemistry. METHODS: Liver biopsies from 15 early (stages I and II) and 14 late (stages III and IV) cases of PBC and 15 normal cases were examined immunohistochemically for expression of p53, CD95/Fas, bax, bcl-x, bcl-2 and the proliferation marker Ki-67. RESULTS: CD95/Fas, bax and bcl-x were identified in biliary epithelium in 8/15, 11/15 and 8/15 normal biopsies. Weak expression of bcl-2 was found, but p53 was not identified. In cases of PBC surviving bile ducts showed strong bax and bcl-x expression. Inflammatory infiltrates were strongly bcl-2 positive. In cases showing a marked ductular reaction there was increased reactivity for bax and bcl-x in ductules. No change in CD95/Fas or p53 expression was seen. An increase in Ki-67 positive biliary epithelial cells was seen in PBC cases, indicating cell cycle activity. CONCLUSIONS: Bile duct epithelium constitutively expresses several genes involved in the execution of apoptosis but these cells also retain the ability to proliferate. PMID- 9855078 TI - Relationships between endosonographic appearan e and clinical or manometric features in patients with achalasia. AB - BACKGROUND: The existence of endosonographic abnormalities of the oesophagus in achalasia is discussed. The place of endoscopic ultrasonography (EUS) needs to be clarified. PATIENTS: Thirty five untreated patients suffering from achalasia and 28 controls without oesophageal disease were prospectively enrolled since 1993. Pseudoachalasia was diagnosed in two patients. METHODS: EUS measurements were performed at two opposite sites at the level of the cardia, and 5 cm and 10 cm proximally, avoiding compression by the water filled balloon. RESULTS: The oesophageal wall and the fourth hypoechoic layer were significantly thicker at the level of the cardia and 5 cm above, with mean differences between patients and controls of 0.37/0.42 mm and 0.16/0.23 mm respectively. No statistically significant correlation could be demonstrated between the thickness of the oesophageal wall or of the fourth hypoechoic layer and weight loss, or the average pressure of the lower oesophageal sphincter. However, a significant inverse relationship was demonstrated between the duration of symptoms and the thickness of the fourth hypoechoic layer. The thickness of the fourth hypoechoic layer was also increased in patients who required only one pneumatic dilatation (P < 0.01). CONCLUSION: The thickness of the oesophageal wall and of the fourth hypoechoic layer appeared to be significantly increased in achalasia patients. However, the slight increase of the mean size (< 0.5 mm) of the muscularis propria suggests that EUS is not helpful in the diagnosis of achalasia. The physiopathological basis of advanced achalasia has to be reconsidered as we demonstrated an inverse relationship between the duration of symptoms and the thickness of the muscularis propria. PMID- 9855079 TI - Effect of prokinetics in children with recurrent nocturnal retrosternal pain. AB - BACKGROUND: Children rarely present with vague complaints of nocturnal recurrent retrosternal or upper-abdominal, epigastric, pain. Dysmotility of the upper gastrointestinal tract, including gastro-oesophageal reflux, might be at the origin of these manifestations. OBJECTIVE: To evaluate the clinical effect of prokinetics (cisapride) in children with recurrent nocturnal retrosternal pain, and to measure the effect of cisapride with a combined oesophageal ambulatory 24 h pH and pressure monitoring. PATIENTS: Twelve children, mean age 9.5 years (range 4.1-14.0 years), with nocturnal recurrent retrosternal pain for more than 6 months, occurring at least three times a week. METHODS: Single-blinded prospective study, with two periods of 2-3 weeks during which cisapride was administered, alternated with two placebo periods of the same duration. Clinical evaluation, and assessment with oesophageal pH and pressure monitoring. RESULTS: Cisapride was related to a temporary relief of the symptoms in 10/12 children. The improvement was demonstrated through a normalization of pH monitoring, and an increase in amplitude and duration of the contractions in the lower oesophagus. CONCLUSION: Cisapride improves complaints of recurrent nocturnal retrosternal pain in children. PMID- 9855080 TI - Validation of [13C]urea breath test for Helicobacter pylori using a simple gas chromatograph-mass selective detector. AB - OBJECTIVE: Isotope ratio mass spectrometry (IRMS) is the accepted method for accurately measuring the 13CO2:12CO2 ratio in the non-invasive and non radioactive [13C]urea breath test (13C-UBT) for Helicobactor pylori. The IRMS instrument, an expensive and highly specialized analyser, is rarely available. The objective of this project was to modify and validate the use of a simple bench-top gas chromatograph-mass selective detector (GC-MSD) for 13C-UBT. METHODS: Breath samples from 71 patients were taken at baseline and 30 min after ingestion of 100 mg [13C]urea. The breath samples were analysed using GC-MSD in the selected ion monitoring mode. The reference 13CO2:12CO2 ratio was from NBS19 obtained from the US National Institute of Standards and Technology. 13CO2:12CO2 ratios of the breath samples were determined. Excess delta per thousand (per mil, delta/thousand) of the 30 min sample over the baseline (deltadelta/thousand) of > or = 6deltadelta/thousand was considered H. pylori positive. Results from 13C-UBT and histology determined blind to each other were compared. RESULTS: The coefficient of variation of the reference 13CO2:12CO2 ratio was 0.06%. Using histology as the 'gold standard', the sensitivity (97.9%) and specificity (95.8%) of the GC-MSD 13C-UBT were comparable to those of other methods of H. pylori diagnosis. CONCLUSION: A gas chromatograph coupled to a mass selective detector that is available in many analytical and biomedical laboratories can be used for the 13C-UBT. This method will increase the availability and reduce the cost of this non-invasive, non-radioactive diagnostic test. PMID- 9855081 TI - Plasma free radical activity and antioxidant vitamin levels in dyspeptic patients: correlation with smoking and Helicobacter pylori infection. AB - BACKGROUND: The pathological processes by which Helicobacter pylori infection leads to the development of gastroduodenal disease are still incompletely understood. Oxygen-derived free radicals are important mediators of inflammation and potential carcinogens. Furthermore, dietary studies have suggested that antioxidant vitamins may protect against gastric cancer. OBJECTIVE: To determine plasma free radical activity and antioxidant vitamin levels in dyspeptic patients and to correlate the results with H. pylori infection and tobacco smoking. SUBJECTS: Forty-three patients undergoing routine endoscopy for investigation of dyspepsia. METHODS: Plasma free radical activity was determined by measurement of thiobarbituric acid-reactive substances (TBARS). Plasma samples were also assayed for the antioxidant vitamins A, C and E. Gastroduodenal biopsies were obtained from all patients for histological examination. RESULTS: Plasma TBARS levels were significantly higher in H. pylori positive versus negative subjects (P < 0.03), smokers versus non-smokers (P < 0.04) and males versus females (P < 0.01). Multiple regression analysis revealed that after correcting for male sex and smoking there was no significant association between plasma free radical activity and H. pylori infection. Smokers had significantly lower levels of plasma vitamin C than non-smokers (P< 0.05); no differences were seen in vitamin A and E levels. Gender and H. pylori infection did not significantly affect plasma antioxidant vitamin levels. Gastroduodenal disease was present in all of the smokers compared with 67% of the non-smokers (P < 0.05); 69% of the smokers were H. pylori positive versus 53% of the non-smokers. CONCLUSIONS: Tobacco smoking and male sex, both recognized risk factors for gastroduodenal disease, appear to be the major determinants of increased plasma free radical activity in dyspeptic subjects, rather than H. pylori infection. The reason for the higher prevalence of H. pylori infection and gastroduodenal disease in dyspeptic smokers is unclear but may relate to weakened antioxidant defences. PMID- 9855082 TI - Eradication of Helicobacter pylori by a 1-week course of famotidine, amoxicillin and clarithromycin. AB - OBJECTIVES: The combination of a proton pump inhibitor (PPI) such as omeprazole with amoxicillin and clarithromycin constitutes one of the most effective treatments for the eradication of Helicobacter pylori. Nevertheless, the mechanisms of interaction between these drugs remain unclear. It has been shown that minimal inhibitory concentration values of both antibiotics are considerably lower at neutral pH levels than in an acid environment. Further, omeprazole possesses bacteriostatic activity. To evaluate the significance of these mechanisms we replaced omeprazole with famotidine, a drug which only suppresses acid production, but has no intrinsic antimicrobial activity. METHODS: We evaluated the efficacy of a 1-week course of famotidine 80 mg b.i.d., amoxicillin 1000 mg b.i.d. and clarithromycin 500 mg b.i.d. in a pilot study (20 patients), and then confirmed our results in a larger replication study (87 patients). A total of 107 patients with H. pylori-associated duodenal ulcer (n = 54), gastric ulcer (n = 14) or non-ulcer dyspepsia (n = 39) were included. Endoscopy was performed at baseline and 4-6 weeks after discontinuation of treatment. H. pylori status was assessed by the urease test and histology. RESULTS: H. pylori was successfully eradicated in 94 of 104 patients who completed the study (90.4%; CI 95%, 83.0-95.3%). By intention-to-treat analysis, the eradication rate was 87.9% (CI 95%, 80.1-93.4%). Ulcer healing was observed in 98.1% of duodenal ulcers and 92.9% of gastric ulcers (based on per-protocol analysis). Mild side effects that did not require termination of treatment were reported by seven patients (6.7%). CONCLUSION: A 1-week course of famotidine, amoxicillin and clarithromycin is a highly effective, simple and safe eradication regimen. Our data indicate that acid suppression is the crucial mechanism by which the activity of amoxicillin and clarithromycin against H. pylori is enhanced, whereas additional antimicrobial activity or other specific effects of PPIs seem to be less important. PMID- 9855083 TI - Serum antibodies to H+,K+-ATPase, serum pepsinogen A and Helicobacter pylori in relation to gastric mucosa morphology in patients with low or low-normal concentrations of serum cobalamins. AB - OBJECTIVES: To compare the diagnostic performance of serum antibodies to H+,K+ ATPase (EC 3.6.1.36), serum pepsinogen A (EC 3.4.23.1) and the Schilling test in diagnosing chronic atrophic body gastritis; to study the interrelationships between H+,K+-ATPase antibodies, serology for Helicobacter pylori, and gastric morphology. DESIGN: Patients with suspected cobalamin deficiency and serum cobalamin < 200 micromol/l were investigated using upper gastrointestinal endoscopy, the Schilling test and serum tests for H+,K+-ATPase antibodies, pepsinogen A, and H. pylori. SETTING: The Department of Internal Medicine, Sahlgrenska University Hospital, Goteborg, Sweden. PATIENTS: Ninety seven consecutively referred patients. MAIN OUTCOME MEASURES: Sensitivity and specificity of assays for serum H+,K+-ATPase antibodies, serum pepsinogen A, and the Schilling test. RESULTS: Assays of serum antibodies to H+,K+-ATPase and of serum pepsinogen A displayed equal diagnostic sensitivity for atrophic gastritis (around 0.90 for the severe forms) and higher than that for the Schilling test (0.65). The diagnostic specificity for pepsinogen A (1.0) was higher than for H+,K+-ATPase antibodies (about 0.80). The prevalence of antral gastritis and positivity for H. pylori antibodies declined with the transition of body gastritis into severe atrophy, while the prevalence of H+,K+-ATPase antibodies increased. CONCLUSION: Pepsinogen A is preferable to serum H+,K+-ATPase antibodies in the diagnosis of gastric body mucosal atrophy. The formation of H+,K+-ATPase antibodies does not seem to be a primary event in the development of gastric body muscosal atrophy. PMID- 9855084 TI - Determinants of prescribing costs for ulcer-healing drugs and upper gastrointestinal endoscopy in general practice. AB - BACKGROUND: Prescriptions for ulcer-healing drugs (UHDs) and endoscopy costs represent major expenditures for dyspepsia in primary care. Healthcare expenditure for dyspepsia could be better understood if the factors contributing to the expenditure for dyspepsia could be identified. AIMS: To determine whether prescribing costs of UHDs and use of endoscopy in general practice were related to the characteristics of the practices or to the characteristics of the population it served DESIGN: Twenty-seven GP practices in south London were studied prospectively over 6 months. Prescribing costs for UHDs were obtained from PACT and data for endoscopies from hospital PAS systems. Demographic data on practice size, age and sex distribution were obtained from the district FHSA. The Jarman index, Townsend score and proportion of ethnic minorities in the practice population were determined from the Population Census Survey. RESULTS: Total expenditure on UHDs by the 27 practices was Pound Sterling 1 million per annum and endoscopy rate was 1.1% per annum. Expenditure on UHDs was negatively correlated with practice size (P = 0.006) and use of open access endoscopy (P < 0.005) and positively correlated with number of patients aged over 45 years (P = 0.007). Endoscopy use was positively correlated with proportion of ethnic minorities (P = 0.008) and negatively with male:female ratio (P = 0.049). CONCLUSIONS: Resource utilization on dyspepsia in general practice is determined by both practice and population characteristics. PMID- 9855085 TI - Intestinal metaplasia in gastric malignancy: a comparison between carcinoma and lymphoma. AB - OBJECTIVES: Intestinal metaplasia is associated with gastric carcinomas, in particular intestinal-type carcinomas. According to the literature, intestinal metaplasia, especially the sulphomucin-rich incomplete type (type IIb or III) can be considered as a precancerous lesion. Intestinal metaplasia is not uncommon in gastric lymphomas, and the purpose of this study was to determine its prevalence and extent, its different subtypes and the distribution of sulphomucin and to compare these findings with those in gastric carcinomas. DESIGN: The prevalence and grade of intestinal metaplasia were investigated in surgical specimens from 160 patients with gastric carcinoma (102) [intestinal type (69), diffuse type (33)] and gastric lymphoma (58) [marginal zone cell (29), diffuse large cell (29)]. Intestinal metaplasia is analysed by specific mucin stains. METHODS: Intestinal metaplasia was described according to the Updated Sydney system. Using periodic acid-Schiff Alcian Blue pH 2.5 and high iron-diamine/Alcian Blue staining, the intestinal metaplasia was classified (Jass classification) and its sulphomucin content was determined. RESULTS: The prevalence and extent of intestinal metaplasia are significantly higher in intestinal-type carcinomas than all other gastric malignancies. Subtyping of intestinal metaplasia is independent of the different types of gastric neoplasia. High sulphomucin positivity in the metaplastic epithelium is only seen in intestinal-type carcinomas, although in a small number of cases (21.5%). CONCLUSION: Prevalence, extent and sulphomucin content of intestinal metaplasia are significantly higher in intestinal-type gastric carcinoma compared to diffuse-type carcinoma, marginal zone cell and diffuse large cell lymphomas in the stomach. PMID- 9855086 TI - Optimization of acid suppression for patients with peptic ulcer bleeding: an intragastric pH-metry study with omeprazole. AB - OBJECTIVE: To study whether an intravenous infusion dose of omeprazole (80 mg + 8 mg/h) during 24 h can be subsequently reduced with maintained effect. Second, to study the effect of oral omeprazole 20 mg given once or twice daily up to day 10, after cessation of a 3-day intravenous infusion (80 mg + 8 mg/h). DESIGN: Prospective, randomized, partly blinded study. METHODS: Twelve Helicobacter pylori(+) patients and 12 H. pylori(-) subjects were included. In part I the patients received omeprazole, 80 mg + 8 mg/h, during 24 h followed by 8, 4 or 2 mg/h. In part II the subjects received 80 mg + 8 mg/h during 3 days followed by 20 mg omeprazole orally, once or twice daily until day 10. Intragastric pH was measured. RESULTS: All H. pylori(+) patients showed a rapid increase of intragastric pH with a mean intragastric pH of 6.7 during the second half of the first day. After the subsequent dose reduction, the mean pH decreased to 6.1-6.2. Patients continuing on 8 mg/h showed the best results. Likewise, all H. pylori(-) subjects showed a rapid and sustained reduction of intragastric acidity during the infusion. Subsequent dose reduction to 20 mg once daily led to a stable fraction of time with pH > 3 of 72%. CONCLUSIONS: Omeprazole given as a continuous infusion of 80 mg + 8 mg/h for 72 h followed by omeprazole 20 mg once daily raised the intragastric pH to and above levels alleged to allow haemostasis in patients with peptic ulcer bleeding and subsequent healing of the ulcer. PMID- 9855087 TI - Mechanisms of diarrhoea in myotonic dystrophy. AB - BACKGROUND: Gastrointestinal (GI) symptoms are common in myotonic dystrophy (MD). Diarrhoea is one of the more disabling of these GI complaints. The mechanisms behind diarrhoea in MD have not previously been investigated systematically. OBJECTIVE: To elucidate the mechanisms behind diarrhoea in MD. METHODS: Twenty patients with MD and suffering from diarrhoea were investigated in order to detect malabsorption (blood tests and faecal fat excretion) and bile acid malabsorption ([75Se]selenahomocholic acid-taurine (SeHCAT) retention) and to study intestinal morphology (duodenal and rectal biopsies). RESULTS: Two patients had deficiency of folic acid and four showed reduced levels of pancreatic isoamylase, but none of them had steatorrhoea. Two out of eight patients had abnormal bile acid breath tests with normal SeHCAT, indicating small bowel bacterial overgrowth and 12 displayed reduced SeHCAT retention. Duodenal biopsies were normal in eight patients and five out of nine rectal biopsies displayed slight inflammation. CONCLUSIONS: A possible mechanism of diarrhoea in MD could be identified in most of the patients. Bile acid malabsorption seems to be a frequent cause and can be treated successfully. PMID- 9855089 TI - Coeliac disease presenting with intraperitoneal haemorrhage. AB - A 42-year-old man, who was previously fit and well, presented in haemorrhagic shock due to a spontaneous left mesocolonic haematoma and intraperitoneal bleed. His INR was noted to be raised on admission. Later investigations showed him to have villous atrophy on biopsy of the second part of his duodenum and a positive anti-reticulin antibody. His duodenal biopsy and INR normalized on a gluten-free diet. Coeliac disease may present with a single vitamin deficiency with potentially catastrophic results. PMID- 9855088 TI - Interleukins and their antagonists but not TNF and its receptors are released in post-ERP pancreatitis. AB - OBJECTIVE: Usually it is not possible to study the initial systemic response in patients with acute pancreatitis in the first hours after onset of the disease. We used postendoscopic retrograde pancreatography (ERP) pancreatitis as a model to study cytokine and anticytokine release in the early phase of human acute pancreatitis. METHODS: Post-ERP pancreatitis was defined as a threefold increase in serum amylase and at least two of the following clinical symptoms: abdominal pain, nausea, vomiting or peritonism 24 h after ERP. Serum levels of pro inflammatory cytokines interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interleukin-8 (IL-8), tumour necrosis factor alpha (TNF), as well as endogenous antagonistic mediators of the systemic inflammatory response such as soluble tumour necrosis factor alpha receptors p55 (TNFR p55) and p75 (TNFR p75), and IL 1-receptor antagonist (IL-1-RA) and interleukin-2-receptor (IL-2R) as indicators of lymphocyte activation were measured before and 0, 1, 4, 12, 24 and 48 h after ERP. In nine patients with acute post-ERP pancreatitis, these parameters were monitored daily until C-reactive protein (CRP) was within normal ranges and were compared to patients without pancreatitis after ERP. RESULTS: IL-1beta was not detectable in five patients with and four patients without post-ERP pancreatitis. The values of the remaining patients in both groups were lower than 3.9 pg/ml. IL 8 and IL-1-RA serum concentrations peaked 12 h after ERP (132.9 and 3245.0 pg/ml respectively) compared to patients without post-ERP pancreatitis (25.8 and 389.9 pg/ml respectively). The IL-6 concentration increased to 81.6 pg/ml (8.0 pg/ml in control patients) 24 h after ERP, while the peak values for CRP were measured 72 h after ERP (164.0 versus 7.7 mg/l). IL-2R content was maximally elevated 144 h after ERP (688.8 versus 255.9 U/ml), while concentrations of TNF and its receptors showed no significant change over time. CONCLUSION: The initial response of the cytokine network to damage of the human pancreas leading to acute pancreatitis includes the release of IL-8 and the IL-1 antagonist IL-1-RA, while IL-1beta is not found in the systemic circulation. The TNF system does not seem to be involved as indicated by the lack of detectable changes in TNF and the soluble TNFR p55 and p75 serum concentrations. Lymphocyte activation as indicated by elevated IL-2R levels occurred days after the initial trauma. Even mild post ERP pancreatitis leads to significant systemic release of cytokines and their biological counterparts. PMID- 9855090 TI - Recurrent cardiac tamponade as first manifestation of gastric cancer. AB - Malignant pericardial effusion is an uncommon disorder and is usually caused by far advanced lung cancer, breast cancer, lymphoma and leukaemia. Pericardial effusion in recurrent gastric cancer has been reported in only three patients. We report the case of a 53-year-old male with sudden onset of dyspnoea, pericardial effusion and cardiac tamponade and the unexpected and the asymptomatic concurrence of gastric cancer. Recurrent haemorrhagic pericardial effusion with physical signs of cardiac tamponade as the initial and only clinical manifestation of gastric cancer has not been described previously. PMID- 9855091 TI - The Fontan type procedure in patients with visceral heterotaxy. PMID- 9855092 TI - Ventricular septal defect: (not) another epidemic? PMID- 9855093 TI - On congenital heart disease in developing countries. PMID- 9855094 TI - Edgar Mannheimer lecture. Termination, consent and innovation: ethical and legal aspects of paediatric cardiology. PMID- 9855095 TI - Pediatric cardiac surgery in Indonesia. AB - Pediatric cardiac surgery in Indonesia first developed thanks to the cooperation of various cardiac centers abroad. The establishment of the 'Harapan Kita' National Cardiac Center in 1985 was one of the most important initial steps. Thereafter, the discipline advanced remarkably in terms of the number of the operations performed and the variety of the diseases treated and, as a result, the surgical outcome also improved. Numerous problems remain to be solved. Only 1% of the children with congenital heart disease are today properly treated in Indonesia. Some of the underlying problems responsible for this situation include a shortage of pediatric cardiac professionals, the lack of the information and education on the part of the patients, and a shortage of funding, both privately and publicly. It would thus be welcome for pediatric cardiac surgeons, cardiologists and nurses in Indonesia to learn about congenital heart disease from doctors and nurses in advanced countries in order to improve the outlook at home. PMID- 9855096 TI - Sterile vegetations in children with severe tetralogy of Fallot. AB - The finding of bland, sterile vegetations in children with severe tetralogy of Fallot is unexpected, and to our knowledge, has not been reported previously. Eight patients diagnosed with tetralogy between January 1993 and July 1997 had sterile vegetations proven by histological and microbiological evaluation, in their right ventricular outflow tracts. Four of these patients were experiencing severe hypercyanotic spells, and four had severely reduced effort tolerance at presentation. They all underwent cardiac catheterization and were submitted for surgical repair. At surgery, the vegetations were thought to be causing further narrowing of the already tight fibrotic infundibular stenosis. Two of these patients had evidence of damaged valves, without evidence of active endocarditis. Although initially sterile, these vegetations, may in some instances, become infected. PMID- 9855097 TI - Superior caval venous syndrome after atrial switch procedure: relief of complete venous obstruction by gradual angioplasty and placement of stents. AB - Superior caval venous syndrome is one of the late problems known to occur after Mustard repair of complete transposition. Reoperation may leave residual stenosis, and carries substantial risk for the patient. It is now feasible to use intravascular stents to overcome systemic venous baffle obstructions, and such an approach is probably more effective. The purpose of our study therefore, was to assess immediate and medium term results of inserting stents subsequent to gradual balloon enlargement of acquired atresia of the intraatrial baffle in patients who had undergone an atrial switch operation. We investigated five patients with complete obstruction of the superior caval venous pathway at perforation of the atretic segment was achieved using a guide wire technique. The procedure was successful in all patients. Gradual angioplasty was performed and intravascular stents were implanted. The pressure in the superior caval vein dropped to normal values, symptoms improved, and the patency of the newly created venoatrial communication was proven at mid-term follow-up. Thus critical obstructions at the superior caval venous pathway after the Mustard procedure can be reopened by interventional catheterization. Implantation of balloon-expandable intravascular stents is safe and effective in the acute relief of the obstructions, but careful long-term follow-up is mandatory. PMID- 9855098 TI - Anomalous origin of one pulmonary artery from the ascending aorta: 36 years' experience from one centre. AB - Anomalous origin of one pulmonary artery from the ascending aorta is an uncommon lesion with an uncertain outcome. We reviewed 16 consecutive children (9 males) presenting with this lesion over a 36 year period at a single institution. Median age at presentation was 2 days (range, birth to 3.2 years). The anomalous pulmonary was the right in 12 and the left in 4, all originating from the proximal ascending aorta, with no patient having stenosis at the origin of the anomalous pulmonary artery. Associated cardiac anomalies were noted in 9 patients. No intervention was attempted in 2 patients: one was deemed inoperable due to complex associated lesions and pulmonary vascular obstructive disease, while the other one died before repair. Surgical intervention was attempted in 14 patients, with 3 intraoperative deaths (21%). Of 11 operative survivors, 8 developed pulmonary arterial stenosis graded severe in 2, moderate in 1 and mild in 5. Patients with severe stenosis required surgical angioplasty, 1 after unsuccessful dilation combined with placement of an endovascular stent. One patient with moderate, and one with mild, stenosis underwent successful transcatheter balloon dilation. The remaining 4 patients with mild stenosis remain unchanged during follow-up. One patient had biopsy evidence of pulmonary vascular obstructive disease at 3.3 years of age. There were no late deaths, giving a total mortality of 25% (4/16). CONCLUSION: While early diagnosis and repair of anomalous origin of one pulmonary artery from the ascending aorta is necessary, restenosis of the site of repair is common. PMID- 9855099 TI - Atrioventricular septal defect with common valvar orifice and tetralogy of Fallot revisited: making a case for primary repair in infancy. AB - Atrioventricular septal defect with common valvar orifice and tetralogy of Fallot is a rare combination of congenital cardiac anomalies. Approaches to this lesion have tended to emphasize either staged repair or complete repair beyond infancy. Between July 1992 and August 1997, nine patients underwent repair of complete atrioventricular septal defect with tetralogy of Fallot. One patient, aged 9.6 years at the time of repair, had previously undergone construction of a modified Blalock-Taussig shunt. Primary complete repair was performed in the other 8 patients at ages ranging from 2.5 to 16 months (median 4.6 months), and all but one were infants. All patients had a Rastelli type C defect, a single ventricular septal defect with inlet and outlet components, and malalignment of the muscular outlet septum with subpulmonary stenosis. A single patch technique, with closure of the zone of apposition ('cleft') in the left atrioventricular valve, was used in all eight patients undergoing primary repair, while a double patch was employed in the previously palliated older patient. In all cases of repair using a single patch, the anterosuperior bridging leaflet was divided obliquely to the right, following the malaligned outlet septum, in order to avoid subaortic obstruction. Repair of the right ventricular outflow tract included infundibular myectomy in eight, pulmonary valvotomy in four, infundibular or transannular patching in three and one, respectively, and reconstruction with a valved allograft conduit in two patients. There was no early mortality or significant morbidity. At a median follow-up of 45 months, there had been one death related to non-cardiac causes and no reinterventions. Left atrioventricular valvar regurgitation was moderate or mild in two patients, and right atrioventricular valvar regurgitation was mild in one patient. No patient had more than mild pulmonary regurgitation or a gradient across the right ventricular outflow tract in excess of 18 mm Hg. Our results demonstrate that primary repair of atrioventricular septal defect with tetralogy of Fallot can be performed with excellent early and mid-term results in young infants. Although it has been suggested that a technique utilizing oblique division of the anterosuperior bridging leaflet may lead to high rates of atrioventricular valvar regurgitation, medium-term atrioventricular valvar function in the present cohort of patients has been excellent. PMID- 9855100 TI - Transcatheter closure of atrial septal defects within the oval fossa: medium-term results in children using the 'ASDOS'-technique. AB - OBJECTIVES: The purpose of this study was to evaluate the safety and efficacy of the ASDOS-technique (Sulzer-Osypka GmbH, Germany) for transcatheter closure of atrial septal defects within the oval fossa. BACKGROUND: Although several attempts have been made to occlude defects within the oval fossa by transcatheter techniques, none of these has gained general acceptance. METHODS: Patients with a defect in the oval fossa measuring equal to or less than 20 mm diameter, with a residual septal rim of 5 mm or greater, body weight greater than 10 kg, with clinical indications for surgical closure were considered for transcatheter closure. Follow-up investigations were performed at discharge, after 1, 3, 6 and 9 months, as well as after 1 and 2 years. RESULTS: Of 78 patients considered for closure, a device was inserted in 41 patients (53%), with success being achieved in 40 patients (98%). The ages ranged from 1.1 to 15 years (7.8 +/- 1.92 years), the 'stretched' diameter of the defect from 10 to 20 mm (14.7 +/- 2.60 mm), and the diameters of the inserted devices from 25 to 45 mm (33.2 +/- 5.43 mm). Transient impairment of atrioventricular conduction occured in 4 patients. During the follow-up of 23.0 +/- 5.6 months elective surgical closure of a residual shunt was performed 26 months after insertion of the device in one patient. None of the other patients required surgery, hospitalisation or medical treatment, and none is requiring further treatment of the defect within the oval fossa. Fracture of one arm of the device occurred in 4 patients, but the fractured arms are in an unchanged and stable position after a period of at least 19 months. CONCLUSIONS: Our medium-term data show that transcatheter closure in children of defects within the oval fossa can be performed with a high efficacy and safety using the ASDOS-device. PMID- 9855101 TI - Congenitally corrected transposition with atrioventricular septal defect. AB - We describe 4 cases of congenitally corrected transposition associated with atrioventricular septal defect, diagnosed by echocardiography and angiocardiography. Two had usual atrial arrangement and two had mirror imaged atrial arrangement . All cases were associated with subpulmonary valvar stenosis. All patients presented with cyanosis and were in sinus rhythm. Atrioventricular septal defect with common atrioventricular junction was easily diagnosed on the basis of a common atrioventricular valve permitting interatrial and interventricular communications. All patients had balanced right and left ventricles. The echocardiographic recognition of the ventricles was based on the presence of the moderator band within the morphologically right ventricle, the characteristics of the apical septal trabeculations, and the shape of the ventricles. Angiocardiographic recognition of the ventricles was achieved on the basis of right and left ventriculography. In one case with usual atrial arrangement, we recorded two His bundle potentials, one anteriorly and another posteriorly. Atrial stimulation revealed blocked atrioventricular conduction at the level of the posterior bundle, and normal atrioventricular conduction through the anterior bundle. In both cases with atrial mirror-imagery, only a posterior His bundle potential was found, with normal atrioventricular conduction revealed by atrial stimulation The clinical course with this combination depends on the other lesions present in addition to the common atrioventricular valve. Our electrophysiological studies show that the conduction system in presence of a common atrioventricular valve is as expected for congenitally corrected transposition with two atrioventricular valves. PMID- 9855102 TI - Extracardiac conduit Fontan for children with heterotaxy and functionally single ventricle. AB - Children with functionally single ventricle in the setting of visceral heterotaxy (isomerism) may present a surgical challenge at the time of Fontan completion because of anomalies of systemic and pulmonary venous drainage. We have used an extracardiac conduit in this population to direct inferior caval venous blood to the pulmonary arteries. Over the past five years, nine children with heterotaxy and a functionally single ventricle underwent correction by placement of an extracardiac synthetic (Gore-Tex) conduit. All patients had previously undergone a bidirectional Glenn procedure. Age and weight at the time of insertion of the extracardiac conduit were 26 +/- 15 months, and 11 +/- 2 kilograms, respectively. RESULTS: Of the nine children, six had an uneventful recovery. One developed elevated venous pressures and required a 'fenestration procedure'. Two patients developed pleural effusions. Median length of stay in hospital was 10 days. All children are alive and well, with follow-up of 19 +/- 16 months. There have been no thromboembolic complications. CONCLUSIONS: The extracardiac conduit has worked well in our experience for the completion of the Fontan circulation in children with functionally single ventricle in the setting of visceral heterotaxy. PMID- 9855103 TI - Results of 546 Blalock-Taussig shunts performed in 478 patients. AB - Between 1983 and 1995, 546 Blalock-Taussig shunt procedures were performed in 472 patients: 128 (23.0%) were classical shunts, 90 of them on the same side as to the aortic arch, and 418 (77.0%) were modified shunts, 182 on the same side of the arch. At the time of surgery, 78 patients were aged below one week, 270 from one week to 12 months, and 198 patients were over one year of age. The mean pre operative arterial saturation (71.7% +/- 16.5%) was significantly increased to 83% +/- 17.9% immediately after the procedure (p=0.017). The overall hospital mortality rate was 2.9% (16/546), with rates of 2.3% (3/128) for the classical, and 3.1% (13/418) for the modified shunts (p=not significant). The rate was significantly higher, however, for classical shunts when the pulmonary arterial diameter was less than 4 mm (15.4% versus zero; p=0.047), though this relationship was reversed for modified shunts (zero versus 3.6%; p=0.338). Early mortality was significantly influenced by the age at surgery, 5/78 (6.4%) in patients aged below 1 week, 3.7% between 1 week and 1 year, and 0.5% over 1 year (p=0.019). Early mortality was also significantly increased in patients weighing 3kg or less, 8/156 (5.1%), versus 3/303 (1.0%), p=0.037. Overall, 51 shunts failed (9.3%), 10 early and 41 late. Early failure was significantly increased in patients weighing 3kg or less, 8/156 (5.1%) versus 3/303 (1.0%), p=0.016. The overall early failure rate was 1.4% (3/215) when heparin was administered intra operatively and for 48 hours postoperatively, in contrast to an early failure rate of 3.4% (7/203) when heparin was not used (p=0.294). Overall rates of failure during follow-up were 9.1% (17/188) in heparinized patients versus 13.6% (24/177), (p=0.173) in non-heparinized patients. Failure of classical shunts was 10.2% (13/128), compared with 6.7% (28/418) for modified shunts (p=0.195). Failure was more common overall if the pulmonary arterial diameter was less than 4 mm, 14.7% (9/61), as opposed to 8.7% (26/300) when the diameter was 4 mm or greater, (p=0.144). Administration of aspirin during follow-up after the modified shunt procedure reduced failure from 11% (18/163) to 6.7% (10/150), p=0.176. Classical or modified Blalock-Taussig shunts, either on the same side or opposite to the aortic arch, can be performed on patients of any age with minimum postoperative complications and low operative mortality. The use of intra- and post-operative heparin appears to reduce the overall rate of failure, and the administration of aspirin during follow-up appears to reduce failure of modified Blalock-Taussig shunts. PMID- 9855104 TI - Covert fears, anxiety and depression in congenital heart disease. AB - This study compared anxiety, fears, depression and behavioural problems as occurring in children with congenital heart disease, comparing them with samples of normal children. It further considered the influence of maternal anxiety, as well as analyzing a subgroup of children with cyanotic forms of congenital heart disease to determine if they were at higher risk than acyanotic children for the problems identified. METHOD: We recruited 40 consecutive children with congenital heart disease without obvious psychosocial problems from the Cardiology clinic at the Alberta Children's Hospital. Of the 40 children, 39 families consented to have the children participate, of which 24 were cyanotic and 15 acyanotic. Children completed the revised versions of the Fear Survey Scale-Revised and the Child Manifest Anxiety Scale as well as the Child Depression Inventory. Mothers completed the Child Behaviour Checklist, and the State Trait Anxiety Scale. RESULTS: Children with congenital heart diseases demonstrated more medical fears, and more physiological anxiety, than the normative samples. More specifically, children with cyanotic forms of congenital heart disease demonstrated more fears of the unknown, physiological anxiety, depression, and delinquent behaviors than the acyanotic children with congenital heart disease. Mothers of the children with cyanotic forms of congenital heart disease scored higher on both the state and trait scales, with higher maternal anxiety correlating with higher anxiety, medical fears and behavioral problems in the child. CONCLUSION: In a clinical setting, children with congenital heart diseases who do not present with psychological adjustment problems are still at risk for covert physiological anxiety, medical fears, depression and behavioral problems. The children with cyanotic malformations represent a subgroup at higher risk for these problems, which may be further exacerbated by increased maternal anxiety. PMID- 9855105 TI - Spontaneous closure of muscular ventricular septal defect identified by echocardiography in neonates. AB - Muscular ventricular septal defects were diagnosed by echocardiography in 97 neonates within 7 days of birth. In 82 of the neonates (84.5%), the defect was solitary, while 15 had multiple defects. The solitary defects was located at mid septal, apical, anterior and inlet locations in 42 (51.2%), 21 (25.6%), 14 (17.1%) and 5 (6.1%) neonates, respectively. Multiple defects occurred in the apical, anterior and mid-septal areas. The diameter of the solitary defects ranged from 1 to 6 mm (2.3 +/- 0.8 mm), while the multiple lesions were 1 to 4 mm in diameter (2.1 +/- 0.8 mm) in 28 instances in which they could measured. It proved possible to follow 79 of the patients for period of 10 to 13 months. The defects closed spontaneously in 56 (84.8%) of 66 patients with a single defect, and in 7 (53.8%) of 13 of those with multiple defects (P<0.05). For the solitary defects, the position and size were factors determining the likelihood and speed of closure. Defects located at the apical septum, or defects larger than 4 mm in diameter, closed slowly and at a later stage. Echocardiography is an useful technique in establishing of natural history of muscular ventricular septal defects encountered in neonates. PMID- 9855106 TI - Pericardial effusions in infants and children: injection of echo contrast medium enhances the safety of echocardiographically-guided pericardiocentesis. AB - Pericardiocentesis is usually an easy and uncomplicated procedure when guided by cross-sectional echocardiography, but an abnormal intracardiac or extrapericardial position of the puncture system can occur, especially in children. Injection of echo contrast medium through the puncture needle is a very sensitive, quick, easy and harmless procedure which can be performed at the bedside in all cases in which doubt remains concerning the location of the needle. Prompt enhancement of signal intensity of the pericardial fluid validates the correct intrapericardial position, and permits visualization of the tip of the needle. Absence of echodense formations excludes an intrapericardial position. We recommend the use of echo contrast medium during pericardiocentesis whenever blood is aspirated through the needle, or if there is any doubt concerning its location. PMID- 9855107 TI - Coarctation of persistent right fifth aortic arch and pulmonary sequestration. AB - A 41-day-old boy was transferred to our department with severe congestive heart failure. Digital subtraction counter current aortography, and antegrade aortography, revealed coarctation of a persistent right fifth aortic arch, stenosis of the origin of an aberrant left subclavian artery, and sequestration of the lower lobe of the right lung. As his heart failure seemed to be caused not only by pressure overload to the left ventricle following coarctation, but also by the volume load associated with the shunting effect of the pulmonary sequestration, we performed transcatheter balloon dilation of the coarctation and the origin of the aberrant subclavian artery, together with embolization of the aberrant pulmonary artery. His heart failure responded dramatically to these procedures. PMID- 9855108 TI - Defective lateralisation in children with congenitally malformed hearts. AB - Defects in lateralization can be studied from the stance of populations, the individual, or the systems of organs within each individual. Unfortunately, and confusingly, the same terms are being applied to each of these situations, but inevitably with different meanings. Thus, there is presently no consensus on how we should use terms such as "heterotaxy" and "situs ambiguus". By far the least ambiguous use of these words is encountered when they are applied to the organs. In fact, each system of organs can accurately and simply be described in terms of its left-right morphology. All those organs which are paired then can be described, when interpreted on the basis of their intrinsic morphology, as being usually arranged, mirror-imaged, or as showing left or right isomerism. Within the heart, these changes are seen only in the atrial segment. The criterion for distinction of rightness or leftness within the atrial segment is the extent of the pectinate muscles relative to the atrioventricular junction. Application of this criterion permits unequivocal recognition of symmetry as opposed to lateralization. The same holds good for the other organs. Within any individual organ, therefore, the situation is neither ambiguous nor heterotaxic. Instead, it is lateralised or symmetrical. Within the individual, in contrast, there may well be discrepancies in the expected disposition of the systems of organs which produces potential ambiguity. To dispel this ambiguity, it is necessary to provide a full catalogue. For example, persons with otherwise normally arranged organs may have left bronchial isomerism. Other persons may have discordance between the thoracic organs, which are usually arranged, and the abdominal organs, which are mirror-imaged, but no evidence of isomerism. Within the population, however, we are unaware of any genetically or environmentally induced syndrome in which all individuals show evidence of mirror-imagery, or of isomerism, or of specific discordance between the systems. In fact, all known syndromes encompass all types of defective lateralization. When attempting to identify the genetic mechanisms for production of the syndromes, therefore, it could be positively misleading to attempt to separate isomerism from other perceived forms of"heterotaxy". Our preference is to consider any deviation from the usual arrangement as heterotaxy, and to specify the specific arrangement of the organs within each malformed individual. PMID- 9855110 TI - The origin and course of coronary vessels: embryological considerations. PMID- 9855109 TI - Lesions produced by radiofrequency ablation in a child. PMID- 9855111 TI - Characterization of two Chinese hamster ovary cell lines expressing the COOH terminal domains of sterol regulatory element-binding protein (SREBP)-1. AB - Sterol regulatory element-binding proteins (SREBPs) regulate transcription of genes encoding enzymes in the cholesterol biosynthetic pathway and the LDL receptor. These proteins are synthesized as membrane-bound precursors and processed to generate the NH2-terminal domains, mature transcription factors. We established two Chinese hamster ovary (CHO) cell lines, CHO-421 expressing the truncated hamster SREBP-1 (amino acids 421-1133) with two transmenbrane domains and CHO-557 expressing another truncated SREBP-1 (amino acids 557-1133) without any transmembrane domains, to investigate the fate of the COOH terminus after cleavage of the NH2-terminal mature SREBP. The cell fractionation experiments revealed that the two proteins, regardless of the absence of transmembrane domains in the SREBP (557-1133), similarly localized in the nuclear envelope and the microsomal membrane fractions, suggesting that these proteins appear to be tightly bound to a membrane protein(s) localizing on the nuclear and endoplasmic reticulum (ER) membranes. Although we predicted that overexpression of the COOH terminal domains, which were thought to be involved in the regulation of SREBP processing, would result in disruption of the SREBP-dependent transcriptional regulation of several genes, the mRNA levels for 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) synthase in these two cell lines were regulated in a sterol dependent manner. Pulse-chase experiments revealed that the SREBP (421-1133) was relatively rapidly degraded (t1/2 = 4-6 hr) and that the intracellular cholesterol level did not affect the half-life time. The degradation of the SREBP (421-1133) was not suppressed by the treatment of a calpain inhibitor, N-acetyl leucyl-leucyl-norleucynal (ALLN), which blocks the proteolysis of some proteins within or near the ER. In CHO-557 cells the SREBP (557-1133) was much more rapidly degraded (t1/2 = 1-2 hr), suggesting that the cytosolic COOH-terminal domain is accessible to the enzymatic attacks from the cytoplasm. Taken together, overexpression of the COOH-terminal domains does not affect the regulation of SREBP processing and the domains are rapidly turned over by the cytosolic proteolytic process distinct from the ALLN-sensitive ER degradative pathway. PMID- 9855112 TI - A protein reacted with anti-vitronectin antibody accumulates in tumors derived from B16F10 melanoma cells. AB - Vitronectin is a cell-adhesive glycoprotein in blood plasma and extracellular matrix. We have examined the participation of vitronectin in experimental metastasis of B16F10 mouse melanoma cells which metastasized to lung. By Western blotting, a protein reacted with anti-rat vitronectin antibody was found to be enriched more in the melanoma parenchyma in the lung than the sum of cultured melanoma cells and normal lung tissue. The protein was similarly abundant in tumors of B16F10 cells grown subcutaneously in mice. By immunofluorescence of the tissues, the signal detected with anti-rat vitronectin was found to be localized at nuclei in melanoma cells metastasized in the lung as well as in tumors just grown subcutaneously. These results indicate that the protein reacted with anti vitronectin antibody is expressed much more in the growing mass of B16F10 melanoma cells in vivo than in vitro. PMID- 9855113 TI - Increased expression of low-affinity NGF receptor in rat retinal Muller cells after ischemia and reperfusion. AB - Low affinity nerve growth factor receptor (p75LNGFR) it is thought to play an important role in recovering damaged nerve. To investigate the possible role of p75LNGFR in transient retinal ischemia, we investigated p75LNGFR gene expression and localization. Using rats under anesthetized conditions, we incised the bulbar conjunctive around the limbus, and then clamped the eyes. A sham operation was performed on the contralateral eyes. Ocular ischemia was maintained for 90 minutes. The p75LNGFR gene expression in ischemic rat retinas was examined by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) at 0, 3, 6, and 72 hours after reperfusion, and the localization of p75LNGFR protein in rat retinas was examined by light and electron microscopic immunohistochemistry. The expression of p75LNGFR gene in ischemic rat retinas increased, as compared with that of the contralateral eyes after 6 hours and 3 days of reperfusion. The p75LNGFR protein increased in the outer plexiform layer and in the outer limiting membrane by immunohistochemical technique. Electron microscopic immunohistochemistry demonstrated that the staining is present in the Muller glial cells. The fact that p75LNGFR gene expression increased in Muller cells after reperfusion suggested that p75LNGFR expression may play a curative role in ischemic injury. PMID- 9855114 TI - Effect of microtubule-associated protein MHP1 on microtubule assembly and cell cycle progression in Saccharomyces cerevisiae. AB - Microtubule-associated proteins (MAPs) promote the assembly of microtubules from purified tubulin in vitro. In order to establish a model system for the investigation of the role of MAPs in microtubule assembly in vivo, we have generated Saccharomyces cerevisiae strains that permit the modulation of the expression levels of MHP1 (MAP-Homologous Protein 1) and of the alpha and beta tubulin genes. Simultaneous overexpression of alpha and beta tubulin results in the accumulation of long aberrant microtubules in interphase, a similar phenotype as was observed in cells overexpressing MHP1. We demonstrate that overexpression of MHP1 in asynchronously growing yeast cultures leads to cell cycle arrest in G2. In cells that overexpress MHP1 and the tubulin genes, a suppression of both the MHP1 and the tubulin overexpression phenotypes can be observed. Progressive induction of alpha and beta tubulin overexpression and constitutive overexpression of MHP1 lead to the formation of long cytoplasmic microtubules more frequently than observed in cells overproducing tubulin or Mhplp individually and the increased microtubule polymerization could be correlated with the increase of a and beta tubulin expression. However, the overexpression of MHP1 did not alter the phenotypes of individual overexpression of a or beta tubulin. These data indicate that Mhplp not only stabilizes microtubules but promotes microtubule assembly in vivo, and suggest that the role of other mammalian MAPs in the promotion of microtubule assembly could be tested in this yeast system. PMID- 9855116 TI - Apoptotic cell death of high polyploid cells in a cultured sarcoma cell line. AB - It is well known that DNA-ploidy is useful independent prognosticator of malignancy. However, the biological significance of polyploid cells and the relation between polyploidy and prognosis is not well understood. We analyzed DNA ploidy by flow cytometry in Meth-A cells (a cultured sarcoma cell line) after treatment with K252a, a protein kinase inhibitor, and showed induction of polyploidization. Apoptotic cell death of the high polyploid cells was verified by flow cytometry, morphological observation and gel analysis of DNA integrity. Expression of tumor-suppressor nuclear protein p53 investigated by immunohistochemistry was increased 10-fold or more in cells with 16C (C = haploid DNA content) relative to cells with 2C, suggesting that the overexpression of p53 was involved in the apoptosis. These results may be of clinical relevance since it has been known that both DNA ploidy and p53 expression have prognostic significance. PMID- 9855115 TI - Stimulative effect of high-level hypergravity on differentiated functions of osteoblast-like cells. AB - The exposure of freshly isolated osteoblasts and osteoblast-like cells to high level hypergravity caused the inhibition of cell growth, elevation of cAMP content, and the stimulation of differentiated functions such as alkaline phosphatase activity, collagen synthesis, and osteocalcin synthesis. Blockage of elevation of cAMP by SQ22536, an inhibitor of adenylate cyclase, resulted in the inhibition of the hypergravity-stimulated alkaline phosphatase activity, indicating that cAMP is the intracellular mediator of this action of hypergravity. H89, an inhibitor of cAMP-dependent protein kinase (PKA), further inhibited the cell growth that was already inhibited by the hypergravity, and further stimulated the alkaline phosphatase activity that was already stimulated by hypergravity. If cAMP acts through the PKA system, H89 should have blocked the changes in cell function effected by the exposure to hypergravity. Therefore the elevated intracellular cAMP by the exposure of hypergravity caused the changes in cell function by a PKA-independent pathway. PMID- 9855117 TI - Prophylactic treatment of superficial bladder tumor. PMID- 9855118 TI - Malignant neoplasm in kidney transplantation. AB - BACKGROUND: The kidney recipient is at a higher risk for cancer than is the general population, although the incidence of neoplasms in general is considered lower in Japan than in Western countries. The cause of this increased risk associated with either transplantation or geography has not yet been established. METHOD: The incidences and sites of malignant neoplasms were analyzed in 285 kidney recipients, who had been followed up for 3007 patient-years. The relationship between immunosuppressive states, the numbers of CD4-positive T lymphocytes, and the presence of malignant neoplasms was studied retrospectively. RESULTS: Eighteen malignant neoplasms were found in 17 of the 285 patients (6%). The malignancies developed in these patients an average of++ 1 26.5 months after transplantation. The incidence was only 3.9% at 10 years, increasing to 13.9% at 20 years. No difference in the time-course incidence was found between azathioprine-based and cyclosporin-based immunosuppressive regimens. The malignancies developed in the digestive organs in more than half of the patients, and were mainly in the liver, colon and rectum, and stomach, with a relatively low incidence of skin cancer and lymphoma. There was only one case of Epstein Barr virus genome found in 5 specimens that were tested. Concerning the immunosuppressive state, CD4-positive T lymphocyte counts were not related directly with malignancies in our series. CONCLUSION: The cumulative incidence of malignancy increased markedly in the second posttransplant decade. The site of cancers in kidney recipients mirrors that of general Japanese malignancies. Our results revealed neither the cause nor predictor for malignancies in kidney transplant patients. PMID- 9855119 TI - Clinical and histopathologic examination of renal allografts treated with tacrolimus (FK506) for at least one year. AB - BACKGROUND: We clinically and pathologically analyzed renal allografts from 1 9 renal transplant patients treated with tacrolimus (FK506) for more than 1 year. METHODS: Twenty-six renal allograft biopsy specimens from 1 9 renal transplant patients who underwent transplantations between 1991 and 1993 were evaluated. Thirteen biopsies were performed from stable functioning renal allografts with informed consent (nonepisode biopsy) and the other 13 were from dysfunctional renal allografts with a clinical indication for biopsy (episode biopsy). RESULTS: The main pathologic diagnoses (some overlap) were acute rejection (AR; n = 4), chronic rejection (CR; n=5), AR+CR (n =4), recurrent IgA nephropathy (n =5), normal findings (n =2), minimal-type chronic FK506 nephropathy (n = 9), and mild type FK506 nephropathy (n = 11). Of the nonepisode biopsies, 7 and 4 biopsies showed minimal-type and mild-type chronic FK506 nephropathy, respectively. Chronic FK506 nephropathy consisted of rough and foamy tubular vacuolization (5 biopsies), arteriolopathy (angiodegeneration of the arteriolar wall; 20 biopsies), focal segmental glomerulosclerosis (4 biopsies) and the striped form of interstitial fibrosis (11 biopsies). The serum creatinine levels of patients in the mild-type chronic FK506 nephropathy group, which included 7 episode biopsies, were statistically higher than those in the minimum-type chronic FK506 nephropathy group (P< 0.001). CONCLUSIONS: This study demonstrates that chronic FK506 nephropathy consists primarily of arteriolopathy manifesting as insudative hyalinosis of the arteriolar wall, and suggests that mild-type chronic FK506 nephropathy is a condition which may lead to deterioration of renal allograft function. PMID- 9855120 TI - Results of transurethral resection plus adjuvant intravesical chemotherapy for superficial bladder cancer. AB - BACKGROUND: We analyzed the results of conservative therapy for superficial bladder cancer to determine the risk factors for recurrence and progression. METHODS: Between May 1984 and February 1997, 111 patients with primary superficial bladder cancer were treated by a transurethral resection with or without intravesical instillation of chemotherapy, or for patients with concomitant carcinoma in situ (CIS), bacillus Calmette-Guerin. We examined the relationship between tumor stage, grade, incidence of concomitant CIS and recurrence-free survival according to pathologic findings and the drugs instilled. RESULTS: The incidence of concomitant CIS in pT1, grade 3 tumors was significantly higher than that in pTa, grade 1 tumors (42% vs. 3%, P= 0.006). The 5-year recurrence-free survival rate of all patients was 73%. There was no significant difference in recurrence-free survival and pathologic stage, tumor grade, presence of concomitant CIS, or drugs used for instillation. However, the recurrence-free survival in patients with > or = 5 tumors was significantly lower than in patients with less than 5 tumors. Of the 111 patients, only 3 patients demonstrated disease progression and underwent a radical cystectomy, while 1 patient with a pT1b, grade 3 tumor developed a tumor in the ureter. No patient died of bladder cancer. CONCLUSION: Our results indicate that the prognosis of superficial bladder cancer patients with a high-stage, high-grade (pT1, grade 3) tumor is favorable when treated by a transurethral resection and intravesical instillation. Bacillus Calmette-Guerin therapy is useful to prevent the recurrence of tumors with concomitant CIS. PMID- 9855121 TI - Neuropathic bladder dysfunction in patients with ossification of the posterior longitudinal ligament. AB - BACKGROUND: Ossification of the posterior longitudinal ligament (OPLL) may cause neuropathic bladder dysfunction due to spinal cord involvement. OPLL, unlike a traumatic spinal cord lesion, progresses insidiously and sometimes affects longer cord segments. As the manifestation of bladder dysfunction may depend on the development of OPLL, we studied the relationship between bladder function and roentgenographic changes in the spinal canals of OPLL patients. PATIENTS AND METHODS: Eighteen surgical candidates (11 males and 7 females, 34 to 85 years old) were studied urodynamically. Sixteen underwent CO2-filling cystometry, uroflowmetry and measurement of their residual urine volume. Cystometry was omitted in the remaining 2 patients. The vertical extent of OPLL and the degree of stenosis in the spinal canal was estimated by x-ray films and CT. RESULTS: The cystometric study revealed detrusor hyperreflexia in 2 patients and areflexic or underactive detrusors in 5 patients. Intermittent flows or considerable amounts of residual urine were also observed in the arefilexia/underactive group. Uroflowmetry showed a normal flow with little residual urine in both patients in whom cystometry was omitted. Bladder sensation was maintained in all patients. The occurrence of abnormal detrusor activity had no relationship to the degree of canal stenosis, while the occurrence of an areflexic or underactive detrusor correlated with the vertical extent of OPLL. CONCLUSION: Although detrusor hyperreflexia is common in an upper spinal cord lesion, attention should also be paid to the development of detrusor underactivity in patients with a wide vertical extent of OPLL. PMID- 9855122 TI - Long-term results of definitive treatment in elderly patients with localized prostate cancer. AB - BACKGROUND: The indication and effectiveness of definitive local treatment for prostate cancer in patients with a limited life expectancy remains to be established. This study is a retrospective analysis of the long-term clinical outcome of elderly patients with localized prostate cancer treated by radiotherapy or a radical prostatectomy. METHODS: From 1982 to 1992, 37 patients with localized prostate cancer, aged 70 years or older, were treated initially by a pelvic lymphadenectomy and then with either external radiotherapy (n = 1 7) or a radical retropubic prostatectomy (n = 20). Lymph node metastasis was negative in all the cases, and no patients received hormonal treatment after the lymphadenectomy. The outcome of all patients was evaluated in June 1997. RESULTS: The 10-year overall and relative survival rates for the radiotherapy group were 27% and 85%, which were not significantly different from the rates of patients in the prostatectomy group (38% and 74%, respectively). The 5-year progression free rates for the radiotherapy group and the prostatectomy group were 63% and 95%, respectively (P= 0.06). CONCLUSION: In elderly patients with localized prostate cancer, the superiority of a radical prostatectomy over radiotherapy was not demonstrated in terms of either overall or relative survival rates, although the progression rate tended to be higher in patients in the radiotherapy group. The indication of definitive treatment in elderly patients should be further studied incorporating a quality of life assessment. PMID- 9855123 TI - Ultrasensitive assay of prostate-specific antigen for early detection of residual cancer after radical prostatectomy. AB - OBJECTIVES: Perhaps the greatest value of PSA determination in the treatment of prostate cancer is in determining persistent disease after a radical prostatectomy. We investigated the ability of an ultrasensitive PSA assay to detect residual prostate cancer in men at risk for recurrence after a radical prostatectomy. METHODS: Using the Immulite third-generation PSA assay (detection limit, less than 0.003 ng/mL), and the standard IMx PSA assay, we determined PSA levels in 205 serum samples serially obtained from 34 men after a radical prostatectomy. The average days from surgery to serum sampling was 430 (range, 63 to 1296). Patients were classified as having nonaggressive or aggressive cancers, based on clinicopathologic findings. A biochemical relapse was arbitrarily defined. RESULTS: All 17 patients with nonaggressive cancers had PSA values of less than 0.02 ng/mL throughout the sampling period. Two of these patients (12%) had 2 or more consecutive PSA increases and were considered as a biochemical relapse. In contrast, 14 (82%) of 17 patients with aggressive cancers fit criteria of a biochemical relapse. All of the relapses were identified within 2 years after surgery. The IMx assay detected only 7 biochemical relapses during the same sampling period. CONCLUSIONS: Using the Immulite PSA assay, relapse detection times may be shortened allowing for most serological recurrences to be detected within 2 years after a radical prostatectomy. Patients with aggressive cancers may require frequent postoperative PSA determinations with a highly sensitive PSA assay which would allow early intervention when treatments for relapse are effective. PMID- 9855124 TI - Urologic morbidity and its influence on global satisfaction with treatment outcome after radical prostatectomy for prostate cancer. AB - BACKGROUND: This cross-sectional observational study examined urologic morbidity and its effects on Japanese patients' global satisfaction with their treatment outcome. METHODS: A questionnaire was mailed to 44 men who had undergone radical prostatectomy for prostate cancer at a mean age of 65.5 years and were free from recurrent symptoms at a mean follow-up of 3.3 years. Most (70%) had pathologic stage C or D1 disease and many (86%) had received adjuvant endocrine therapy. After reporting urinary symptoms, the patients estimated their overall urinary, sexual, physical, psychological, and social functioning, and global satisfaction level with their treatment outcome. Regression analysis was performed to detect significant factors in predicting overall urinary conditions or satisfaction. RESULTS: Daily urinary leakage and forceless urinary stream was noted in 30% and 23% of patients, respectively. Overall urinary conditions were evaluated as good or very good in 61 % of patients, and were most significantly associated with the patient's satisfaction with their force of urinary flow (P < 0.001). Global satisfaction with treatment outcome was estimated as good or very good in 80% of patients, and it was predicted by physical functions (P= 0.013) and psychological distress (P= 0.036), and to a lesser extent by urinary conditions (P= 0.195). CONCLUSION: A forceless urinary stream was the most significant determinant for overall urinary conditions in patients who had a radical prostatectomy. Global satisfaction with treatment outcome was only marginally affected by urinary conditions. Physical function and psychological distress were major factors affecting the satisfaction level in this population. PMID- 9855125 TI - Treatment of metastatic nonseminomatous germ cell tumors of the testis: significance of the international consensus prognostic classification as a prognostic factor-based staging system. AB - BACKGROUND: We reviewed treatment results in patients with metastatic nonseminomatous germ cell tumors of the testis and examined the significance of the International Consensus Prognostic Classification to make appropriate risk based decisions concerning induction chemotherapy. METHODS: We divided 37 patients treated with platinum-based combination chemotherapy into good, intermediate, and poor prognostic groups utilizing the International Consensus Prognostic Classification. The data was analyzed for both overall survival and progression-free survival among the 3 prognostic groups. RESULTS: Among the 37 patients, 10 died (8 of progressive disease, 1 of pneumonia during induction chemotherapy and 1 of cyclophosphamide-induced hemorrhagic cardiomyolitis during salvage chemotherapy). The survivors were followed for 6 to 1 84 months from the beginning of induction chemotherapy (median, 80 months). Five of the 37 patients (14%) were classified as having a good prognosis, 1 8 (48%) as intermediate, and 14 (38%) as having a poor prognosis. The patients in the poor prognostic group had a 5-year overall survival of only 40%, while those in the good and intermediate groups had 5-year overall survivals of 100% and 94%, respectively. When we applied the International Consensus Prognostic Classification to patients with advanced disease classified by the Indiana University Staging System, these patients could be clearly divided into good-risk and poor-risk groups. CONCLUSIONS: The International Consensus Prognostic Classification is easily applicable and accurate for risk assessment in patients with metastatic nonseminomatous germ cell tumors of the testis. This classification will now be widely used in general oncology practices and for clinical trials in these patients. PMID- 9855126 TI - Surveillance study for clinical stage I testicular seminomas and nonseminomatous germ cell tumors. AB - BACKGROUND: Optimal therapy for stage I testicular tumors is still controversial. This study evaluated the efficacy of a surveillance policy for patients with testicular stage I seminomas and nonseminomatous germ cell tumors (NSGCT). METHODS: From 1984 to 1996, 24 patients with stage I seminoma and 20 with stage INSGCT were followed after radical orchiectomy with tumor markers and imaging studies. All patients were followed for at least 2 years except for those who recurred within 2 years. Recurrent patients were treated with cisplatin-based chemotherapy. RESULTS: The median follow-up periods for seminoma and NSGCT patients were 41 and 54 months, respectively. Recurrences were detected in 2 seminoma (8.3%) and 10 NSGCT (50%) patients. Eleven of the 12 recurrent patients (92%) were detected within 2 years after orchiectomy. The seminoma patients both recurred in the retroperitoneal lymph nodes, while 70% of the NSGCT patients recurred in the lung and/or retroperitoneal lymph nodes. The recurrent seminoma patients were treated with chemotherapy and are alive without disease for 1 7 and 24 months afterorchiectomy. One NSGCT patient died of cancer, but the other 9 recurrent NSGCT patients are alive without disease at 25 to 113 months after orchiectomy. CONCLUSIONS: Surveillance alone is reliable for monitoring patients with stage I testicular seminoma and NSGCT. The majority of recurrences occurred within 2 years, necessitating intensive follow-up for 3 years. As the lung metastatic rates in NSGCT patients were high, a more accurate assessment for lung metastasis is desirable in these patients. PMID- 9855127 TI - Optimal conditions to induce rat adrenal medullary changes, such as pheochromocytoma, by nicotine. AB - BACKGROUND: Nicotine-induced adrenal medullary hypertrophy in rats has been suggested to be a model for pheochromocytoma. Study conditions for proving such an assertion, however, have not been optimized. METHODS: Studies on strain difference, dose dependency, and the time course of catecholamine metabolism in response to nicotine treatment were conducted. Under the putative optimal experimental conditions, metabolic and histologic changes in the adrenal medulla were investigated. RESULTS: Male Wistar rats treated with a maximum dose of 4 mg/kg per day of nicotine for 9 weeks, including a 2-week lead-in period, developed highly consistent changes in the adrenal medulla. Concerning metabolic indices, the norepinephrine content of the adrenal and the urinary excretion of epinephrine and metanephrine were significantly elevated. Hyperplastic and hyperactive states of the adrenal medulla were also indicated by a morphometric analysis on electron microscopic figures. These showed an enlarged cytoplasmic area, the development of a rough-surfaced endoplasmic reticulum, and an increased number and density of intracellular catecholamine granules. The metabolic changes were found to reverse 3 weeks after the cessation of nicotine administration. CONCLUSION: These results provide better-defined experimental conditions for an animal model of pheochromocytoma. PMID- 9855128 TI - Effects of etidronate disodium on crystallizations in synthetic urine and calcium oxalate crystal adhesion to Madin-Darby canine kidney (MDCK) cells. AB - BACKGROUND: Several reports in the 1970s suggested that etidronate disodium might be clinically useful to prevent calcium stones, but the use of etidronate in the urolithiasis field was discontinued due to adverse effects of this drug on skeletal turnover and mineralization. Because the drug might affect not only crystallization, but also crystal-tubular interactions, we investigated the minimum dose of etidronate necessary to effectively prevent stone recurrence without adverse side effects. METHODS: We examined the effect of etidronate on the crystallization of calcium oxalate, calcium phosphate and magnesium ammonium phosphate using synthetic urine and measured by an aggregometer. We also studied its effect on the adhesion of calcium oxalate monohydrate crystals to Madin-Darby canine kidney (MDCK) cells in vitro. RESULTS: Etidronate affected the crystallization+ of not only calcium phosphate and calcium oxalate, but also magnesium ammonium phosphate in synthetic urine. The inhibitory activities on these crystallizations were detected at extremely low drug concentrations. Etidronate also had a strong inhibitory activity against the adhesion of calcium oxalate crystals to MDCK cells. CONCLUSION: Although further studies are necessary regarding the effects of etidronate on crystallization and crystal adhesion both in vivo and in vitro, and the appropriate schedule of dosing to prevent side effects, it is possible that etidronate may be useful in the treatment of urinary stones. PMID- 9855129 TI - Effect of naloxone on the bladder activity of rabbits with acute spinal injury. AB - BACKGROUND: Naloxone enhances bladder activity in patients with chronic spinal cord injury. However, there are few reports on naloxone for bladder morbidity in acute spinal cord injury. METHODS: We performed a prospective, controlled study of the effects of naloxone on bladder function in rabbits with and without surgical transection of the spinal cord at the 10th thoracic vertebra. Acute and chronic stages of injury were defined according to bladder function. Naloxone was given intravenously at both stages, and intrathecally at the acute stage. Bladder activity was monitored by cystometry. Blood concentrations of methionine enkephalin were measured by radioimmunoassay. RESULTS: Spinal cord injuries were acute 1 or 2 days after surgery, and chronic after 1 or 2 weeks. Bladder capacity significantly decreased after 0.01 mg of intravenous naloxone in uninjured control rabbits, and after 0.03 mg of intravenous naloxone in rabbits with chronic-phase injuries. During the acute-injury phase, 0.3 mg of intravenous naloxone, or 0.02 mg of intrathecal naloxone, was necessary to evoke the micturition reflex. No significant changes in blood enkephalin levels were seen before or after spinal cord injury. CONCLUSION: In rabbits with acute spinal cord injury, intrathecal naloxone evoked the micturition reflex at a much lower dose than did intravenous naloxone. Intrathecal naloxone promises to become a new therapy for the acute stage of spinal cord injury for active recovery of bladder function, and could replace current therapy. PMID- 9855130 TI - Antiproliferative effect of calcitriol on human prostatic cancer cell lines: unrelated to the expression of major histocompatibility complex antigens or intercellular adhesion molecule-1 (ICAM-1). AB - BACKGROUND: In addition to its role in calcium and phosphorus metabolism, calcitriol (1,25-dihydroxyvitamin D3; 1,25-D3) demonstrates multiple effects on cell proliferation/differentiation by expressing major histocompatibility complex (MHC) antigens and intercellular adhesion molecule-1 (ICAM-1). It has recently been reported that 1,25-D3 inhibits the growth of prostatic cancer (PCa) cells. In this study we examined the effect of 1,25-D3 on both the growth and expression of HLA-ABC, HLA-DR and ICAM-1 antigens in PCa cells. METHODS: Four human PCa cell lines (PC-3, PPC-1, ALVA-41 and ALVA-101) were examined. The cell numbers were enumerated, and the effects of interferon-y (IFN-gamma) and 1,25-D3 on the expression of HLA-ABC, HLA-DR and ICAM-1 were quantitated by flow cytometry. RESULTS: A dose-dependent antiproliferative effect of 1,25-D3 was found in all PCa cells lines except ALVA-41.1,25-D3 was approximately 10 times as potent as its analogue 24,25-dihydroxyvitamin D3 in inhibiting the growth of PC-3 cells. Also, the relative inhibitory ability of these compounds paralleled the strength of their binding affinities for the 1,25-D3 receptor, indicating that the antiproliferative effect may require a receptor-ligand interaction. HLA-ABC was expressed in PC-3, ALVA-41 and ALVA-101, but not in PPC-1 cells, while HLA-DR was not expressed on any of the tested cells. IFN-gammacould enhance or induce HLA ABC but not HLA-DR expression in the tested cells. ICAM-1 was expressed in all cells and slightly upregulated by IFN-gamma. CONCLUSION: In this study 1,25-D3 had an antiproliferative effect on 3 of the 4 examined PCa cell lines. PMID- 9855131 TI - Laparoscopy-assisted radical nephrectomy for renal cell carcinoma in patients on long-term hemodialysis: report of 2 patients. AB - We successfully performed a laparoscopy-assisted radical nephrectomy for renal cell carcinoma in 2 patients on long-term hemodialysis. Both tumors were incidentally discovered on screening by abdominal CT scanning. There were no complications during the operation or in the postoperative period, and both patients resumed normal activities by the fifth postoperative day. A laparoscopic assisted radical nephrectomy may be useful for the treatment of renal cell carcinoma in hemodialysis patients. PMID- 9855132 TI - Renal adenoma associated with renal cyst formation. AB - We report a renal adenoma associated with renal cyst formation in a 49-year-old male. A small renal mass inside a right renal cyst was discovered incidentally by ultrasonography. A right nephrectomy was performed. A pathologic examination revealed a well-differentiated, benign papillary tumor which was composed of slightly eosinophilic cells. We therefore diagnosed this patient as having a renal adenoma with hemorrhage. PMID- 9855133 TI - A case of hydronephrosis caused by arteriosclerotic compression of the renal pelvis. AB - We present the case of a 65-year-old woman with vascular compression of the renal pelvis, causing calyectasis. The diagnosis was a subtype of Fraley's syndrome. The cause of compression was concluded to be arteriosclerotic change of both the renal artery and the abdominal aorta. PMID- 9855134 TI - Ureterocolic fistula secondary to colonic diverticulitis. AB - We describe the case of a 45-year-old woman with a ureterocolic fistula caused by colonic diverticulitis. She had a 10-year history of intermittent left flank pain that had not been treated. The fistulous tract between the left ureter and sigmoid colon was confirmed by retrograde urography and a barium enema. A nephroureterectomy was successfully performed. PMID- 9855135 TI - Squamous cell carcinoma in an ileal conduit. AB - We report a patient with squamous cell carcinoma that developed at the ureteroileal anastomosis and extended into the ileal conduit 11 years after a radical cystectomy for transitional cell carcinoma of the bladder. To our knowledge, this is the first report to document the development of a squamous cell carcinoma in an ileal conduit after a radical procedure for bladder cancer. PMID- 9855136 TI - Synchronous testicular seminoma and adrenocortical carcinoma: a case report. AB - We report a rare case of synchronous testicular seminoma and adrenocortical carcinoma. A 57-year-old man had a left testicular seminoma (clinical stage IIIB) with metastases to the lung and paraaortic lymph node. A complete response was obtained after 3 courses of chemotherapy with single-agent carboplatin. However, a left adrenal tumor was detected 1 2 months later and demonstrated a tumor volume doubling time of 2.1 months. Chemotherapy with bleomycin, etoposide and cisplatin failed to stop the tumor growth. A laparoscopic adrenalectomy was performed and pathology revealed an adrenocortical carcinoma. The patient has been free of recurrence for 42 months postoperatively. PMID- 9855137 TI - Terson syndrome. PMID- 9855138 TI - Feeder vessel photocoagulation for subfoveal CNV. PMID- 9855139 TI - Feeder vessel photocoagulation for subfoveal CNV. PMID- 9855140 TI - Traumatic hyphema and patient outcomes--an oversight. PMID- 9855141 TI - Laser thermal keratoplasty for PRK overcorrection. PMID- 9855142 TI - Consumer-driven health care: the world turned upside down? PMID- 9855143 TI - Endophthalmitis after cataract surgery: risk factors relating to technique and events of the operation and patient history: a retrospective case-control study. AB - OBJECTIVE: This study aimed to investigate the relationship between postoperative endophthalmitis and current changes in the cataract operative technique. DESIGN: A retrospective case-control study. PARTICIPANTS: The 22,091 cataract operations performed from 1990 through 1993 at St Eriks Hospital formed the basis for this investigation. In a random fashion, 220 control subjects were selected to be compared with the endophthalmitis cases. MAIN OUTCOME MEASURES: Numerous variables pertaining to the cataract extraction procedure and to the ocular and general health of patients with cataracts were analyzed regarding the development of postoperative intraocular infection. Patient age, presence of diabetes or immunosuppression, type of cataract extraction and intraocular lens (IOL), and intraoperative or postoperative complications were the principal variables assessed. RESULTS: Fifty-seven patients with endophthalmitis were diagnosed, resulting in an overall frequency of 0.26%. Immunosuppressive treatment (P = 0.019), wound abnormality (P = 0.03), and the use of IOLs without a heparinized surface (P = 0.0023) were the only significant risk factors found in a logistic regression model. CONCLUSIONS: The results suggest that cataract operating practice may alter the risk for endophthalmitis in that implanting a heparinized IOL and creating a tight section both seem to provide protection against this dreaded complication. Regarding patient history, an increased susceptibility was found among subjects treated with immunosuppressants. Designing a prophylactic protocol that protects against endophthalmitis more efficiently than did the study prophylaxis of 20 mg of subconjunctival gentamicin, is important not only for this patient subgroup but also for the cataract operated population at large. PMID- 9855144 TI - Predictability of spherical photorefractive keratectomy for myopia. AB - OBJECTIVE: This study aimed to examine the effects of purely spherical excimer laser photorefractive keratectomy (PRK) for myopia. DESIGN: Consecutive case series. PARTICIPANTS: A total of 3218 eyes with a mean preoperative mean spherical equivalent (MSE) of -3.75 diopters (D) +/- 1.73 D standard deviation (SD) (range, -1.00 D to -11.88 D) underwent PRK with a Nidek EC-5000 excimer laser. Eyes were divided into groups based on the degree of preoperative myopia in 1 D steps. INTERVENTION: All eyes underwent PRK with a Nidek EC-5000 excimer laser. MAIN OUTCOME MEASURE: Visual and refractive outcome of PRK treatment was measured. RESULTS: After a mean follow-up period of 52.6 weeks (range, 26-150 weeks), the final MSE was -0.07 D (+/-0.68 D) (range, -5.50 D to +4.50 D). Of the 3218 eyes, 2919 (90.7%) were within 1.00 D of emmetropia, and 3038 (94.4%) of eyes had an uncorrected visual acuity of 20/40 or better, with 1886 (58.6%) achieving 20/20 or better visual acuity. Eyes in the lower preoperative myopia groups had a greater chance of attaining 20/40 unaided visual acuity than those in the higher groups (e.g., 98.3% of the -2D group and 53.6% of the -9D group achieved 20/40 unaided visual acuity). Overall, mean postoperative haze was 0.29+/-0.39 SD (scale, 0-3), and 29 eyes (0.9%) lost 0.3 or more logarithm of the minimum angle of resolution (LogMAR) unit of best-corrected visual acuity. CONCLUSIONS: Excimer laser PRK is an effective treatment for myopia of up to 9.00 D. The outcome parameters are less predictable for eyes with greater than 9.00 D of myopia. PMID- 9855145 TI - Magnification characteristics of fundus imaging systems. AB - OBJECTIVE: To compare the magnification properties of 11 different fundus cameras (including 1 stereo fundus camera), a Rodenstock (infrared) scanning laser ophthalmoscope (SLO), the Heidelberg Laser Tomographic Scanner (LTS), and the Heidelberg Retina Tomograph (HRT). DESIGN: A cross-sectional study of the relationship between the true size of a fundus feature and its photographic computer image in 14 different fundus imaging devices. This relationship was evaluated for each instrument using a model eye adjusted for axial ametropia between +11 diopter (D) and -14 D. To simulate refractive ametropia, the "crystalline lens" was removed to render the model eye aphakic, and the axial length was adjusted to give aphakic ametropia from emmetropia to +20 D. MAIN OUTCOME MEASURES: A correction factor (p) was calculated for each instrument, which can be used in calculations for determining true retinal size. RESULTS: The following were found to be of telecentric construction, Zeiss Oberkochen (WS240 Heidelberg), Zeiss Oberkochen (UK), Zeiss Oberkochen (Cologne), Nikon NF505, Kowa RCXV, SLO prototype (UK), LTS, and the HRT, and each exhibited a constant relationship between p and degree of ametropia of the model eye. The Canon CF6OU, Canon CF6OS, Canon CR4-45NM, Nidek 3-DX, Olympus GRCW, and Carl Zeiss Jena Retinophot were found not to be telecentric and exhibited a linear relationship between p and degree of ametropia of the model eye. For all instruments, p remained unchanged for axial and refractive ametropias of the same degree. CONCLUSIONS: The study has shown that not all fundus imaging systems are telecentric, so the use of a single magnification correction value may not be appropriate. These findings have important implications for the way in which true retinal size calculations are performed. Examples are given to show how the tabulated values of correction factors can be used for both telecentric and nontelecentric cameras in image size calculations. PMID- 9855146 TI - Reliability of intraocular pressure measurements after myopic excimer photorefractive keratectomy. AB - OBJECTIVE: To determine the reliability of intraocular pressure (IOP) measurements by Goldmann applanation tonometry and pneumotonometry in eyes treated with excimer myopic photorefractive keratectomy (PRK). DESIGN: A prospective case series. PARTICIPANTS: Forty consecutive eyes treated with PRK were evaluated. INTERVENTION AND MAIN OUTCOME MEASURES: Central and peripheral corneal Goldmann tonometry and pneumotonometry measurements were done before surgery, at 1 week, and at 1 and 3 months after surgery. RESULTS: The IOP by Goldmann tonometry from the central cornea was significantly lower than the peripheral IOP; however, there was no difference between IOP measured from central and peripheral corneas by pneumotonometry, which, in turn, correlated with peripheral Goldmann measurements. There was a trend, but not a statistically significant correlation, between the spherical equivalent of the treatment and the amount of decrease in central Goldmann IOP. CONCLUSIONS: Pneumotonometry measures the IOP reliably after PRK from all parts of the cornea, whereas central Goldmann tonometry underestimates the IOP by 2.40+/-1.23 mmHg. PMID- 9855147 TI - Natural history of corneal topography after excimer laser photorefractive keratectomy. AB - PURPOSE: To investigate the natural history of corneal topography after excimer laser photorefractive keratectomy (PRK). DESIGN: A prospective, single center clinical study. PARTICIPANTS: A total of 40 eyes of 34 patients with myopia were studied. INTERVENTION: Excimer laser photorefractive keratectomy and computer assisted videokeratography were performed. MAIN OUTCOME MEASURES: Preoperative, 1 week, and 1, 2, and 3 months postoperative topography patterns were compared and changes assessed by averaging defined sectors of the ablation zone in individual maps to produce composite "average" topography maps. RESULTS: Corneal topography was generally not homogeneous at 1 week after PRK. By 3 months, there had been considerable smoothing of corneal contour. A general "central island" effect early in the postoperative period flattened over time. When right and left eyes were evaluated independently, a tendency toward a keyhole-semicircular pattern was seen on average; the maximum flattening was nasal and the least flattening was inferotemporal for both right and left eyes. CONCLUSIONS: A tendency, on average, toward central island and keyhole-semicircular patterns is seen early in the postoperative course after PRK. Central islands tend to evolve into the keyhole-semicircular pattern, and the corneal topography in general after PRK tends to smooth considerably with time. PMID- 9855148 TI - Ophthalmic manifestations of fibrous dysplasia: a disease of children and adults. AB - OBJECTIVE: To determine whether adults constitute a significant proportion of patients presenting with ophthalmic complications of fibrous dysplasia. DESIGN: A retrospective case series. PARTICIPANTS: Twenty patients with biopsy-proven fibrous dysplasia participated. MAIN OUTCOME MEASURES: Patient demographics, major signs and symptoms, occurrence of visual loss, pattern of bone involvement, and radiographic appearance of fibrous dysplasia were studied. RESULTS: In contrast to observations made in the past, the authors found that adults constitute a significant proportion of patients suffering from the ophthalmic complications of fibrous dysplasia. Of 20 patients, 9 were younger than 18 years of age (children) and 11 were 18 years of age or older (adults) at the time of presentation. Changes in facial contour and symmetry were the most common presenting signs and symptoms. Five patients, four of whom were adults, presented because of acute visual loss. Most patients had monostotic lesions that crossed suture lines to involve multiple craniofacial bones. On computed tomographic scan, most fibrous dysplasia lesions had a characteristic, pagetoid appearance, with alternating areas of radiolucency and radiodensity. CONCLUSIONS: Because fibrous dysplasia of the orbital bones can be a cause of significant dysfunction and disfigurement, as well as a treatable cause of blindness in both children and adults, the diagnosis of fibrous dysplasia should not be ruled out based solely on the age of the patient. The characteristic radiologic appearance of this disease allows one to differentiate fibrous dysplasia from other tumors associated with bony expansion or density changes, specifically meningioma. PMID- 9855149 TI - Presumed isolated inflammation of the superior oblique muscle in idiopathic orbital myositis. AB - OBJECTIVE: To describe isolated involvement of the superior oblique muscle in association with idiopathic orbital myositis. DESIGN: A case report with review of the literature. PARTICIPANT: A 57-year-old man with acquired strabismus participated. INTERVENTION: Oral steroid therapy was administered. MAIN OUTCOME MEASURES: Clinical and radiographic features before and after treatment were measured. RESULTS: Computed tomographic scanning showed isolated enlargement of the superior oblique muscle consistent with orbital myositis. Clinical and radiographic abnormalities quickly improved after oral steroid therapy. CONCLUSIONS: Orbital pseudotumor may present as a myositis isolated to the superior oblique muscle. This extremely rare condition has been reported in only three patients, each of whom lacked early diagnosis or treatment. Early recognition may improve chances for a successful clinical outcome. PMID- 9855150 TI - Oncocytoma of the eyelid: an aggressive benign tumor. AB - OBJECTIVE: The authors report the case of an 83-year-old patient with a benign oncocytoma of the inferior eyelid. DESIGN: INTERVENTIONal case report. INTERVENTION: Treatment consisted of a large orbital exenteration followed by reconstruction with a pedicled temporalis muscle flap. MAIN OUTCOME MEASURES: Histologic evaluation and clinical follow-up were measured. RESULTS: After a year of follow-up, there was no sign of local recurrence. CONCLUSIONS: Oncocytomas, even if benign, must be considered as very aggressive tumors and treated accordingly. PMID- 9855151 TI - Intraocular pressure as a risk factor for visual field loss in pseudoexfoliative and in primary open-angle glaucoma. AB - OBJECTIVE: To analyze the relationship between intraocular pressure (IOP) and visual field loss in patients with primary open-angle glaucoma (POAG) and in those with pseudoexfoliative glaucoma (PEXG). DESIGN: A cross-sectional, observational study. PARTICIPANTS: Thirty-one patients with PEXG and 31 patients with POAG that was newly diagnosed were included in this study. MAIN OUTCOME MEASURES: The authors recorded the untreated IOP and the amount of the visual field loss, at presentation, in both study groups. RESULTS: The authors found a significant relationship between IOP and visual field mean deviation (MD) index (P = 0.0001, r = 0.68) in PEXG but not in POAG eyes (P = 0.7). CONCLUSION: The authors found that untreated IOP levels can explain the amount of visual field loss, as measured by the MD index, much better in patients with PEXG than in comparable patients with POAG. Thus, vulnerability of the optic nerve head to increased IOP appears to be different in these two diagnostic categories. PMID- 9855152 TI - Immediate argon laser peripheral iridoplasty as treatment for acute attack of primary angle-closure glaucoma: a preliminary study. AB - OBJECTIVE: This study aimed to examine the intraocular pressure (IOP)-lowering effects and safety of immediate argon laser peripheral iridoplasty (ALPI) as a first-line treatment for acute primary angle-closure glaucoma (PACG). DESIGN: A prospective cohort study. PARTICIPANTS: Ten consecutive patients with their first attack of PACG, with IOP of 40 mmHg or greater, were recruited into the study. INTERVENTION: On presentation, each patient received topical pilocarpine (4%) and timolol (0.5%) and immediate ALPI as primary treatment. The IOPs at 15, 30, and 60 minutes after ALPI were documented by applanation tonometry. When the corneal edema had settled, laser peripheral iridotomy was performed as a definitive treatment. MAIN OUTCOME MEASURES: The IOP, corneal edema, and complications from ALPI were measured. RESULTS: The mean IOP of this group of patients was reduced from 59.5+/-10.4 mmHg to 28.7+/-14.9 mmHg at 15 minutes, 21.7+/-13.1 mmHg at 30 minutes, and 16.0+/-9.4 mmHg at 60 minutes after ALPI. No complications from the laser procedure were encountered during the study period. In nine of the ten patients, the corneal edema cleared up 1 hour after ALPI. In the remaining patient, the cornea cleared up 2 hours after ALPI. CONCLUSION: From this preliminary study, immediate ALPI, without adjunctive systemic antiglaucoma treatment, appeared to be very effective in controlling the IOP and returning corneal clarity in acute PACG. Its safety also appeared reassuring and did not have the risks associated with conventional systemic therapies. PMID- 9855153 TI - Modified aqueous drainage implants in the treatment of complicated glaucomas in eyes with pre-existing episcleral bands. AB - OBJECTIVE: Control of intraocular pressure (IOP) after retinal surgery with scleral band placement may be problematic. In this study, outcome after silastic drainage implant placement is reviewed. DESIGN/PARTICIPANTS: A retrospective chart review of 11 eyes in 11 patients with severe glaucoma after multiple surgeries, including scleral buckle procedures. INTERVENTION: Patients received silastic drainage implant surgery, either a long Krupin Denver valved implant (7) or a trimmed Baerveldt seton (4). MAIN OUTCOME MEASURES: Vision, IOP, and need for further glaucoma medication or glaucoma surgery were measured. RESULTS: One- and 2-year life-table success rates (either complete or qualified success) were 82% and 73%, respectively, defining success as final IOP of 21 mmHg or less without need for further glaucoma surgery and no loss of visual acuity. Half of the successful eyes required antiglaucoma medication. CONCLUSIONS: Silastic drainage implants may be a viable option in the treatment of complicated glaucoma after retinal surgery. PMID- 9855154 TI - Occlusive ligature and standardized fenestration of a Baerveldt tube with and without antimetabolites for early postoperative intraocular pressure control. AB - OBJECTIVE: To determine the effectiveness of a surgical modification for a nonvalved aqueous tube shunt in controlling intraocular pressure (IOP) in the early postoperative period. The effect of antimetabolite use on IOP also was studied. DESIGN: A retrospective study of consecutive patients who underwent modified Baerveldt 350-mm2 implant with varied, nonrandomized, exposure to antimetabolites. PARTICIPANTS: Fifty-one eyes of 46 patients with uncontrolled glaucoma were examined. INTERVENTION: Identical surgical modification of a Baerveldt 350-mm2 tube was performed in all cases and consisted of placement of an occlusive 7-0 polyglactin suture just anterior to the plate followed by a through-and-through penetration of the tube just anterior to the occlusive ligature with a standardized 15 degrees blade. Seventeen eyes were not exposed to antimetabolite, while 2 groups of 17 eyes had 3 minutes' episcleral exposure to either 5-fluorouracil 50 mg/ml or mitomycin C 0.4 mg/ml at the location corresponding to the fenestration. The Tenon's layer and conjunctiva were not exposed because of concerns regarding conjunctival erosion over the device. MAIN OUTCOME MEASURE: Intraocular pressure and number of antiglaucoma medications required were analyzed. RESULTS: For the group, mean IOP before surgery and on postoperative days 1, 4, 10, 21, 42, 63, 84, and 112 was (in millimeters of mercury) 34.6, 20.1, 17.0, 17.2, 22.0, 17.3, 18.7, 17.4, and 15.6, respectively. There was an elevation of IOP at day 21 relative to fibrotic blockage of the fenestration before suture autolysis. This was temporized with antiglaucoma medication until suture autolysis occurred or treated with laser suture lysis (8 eyes). On day 1, hypotony occurred in 3 (6%) eyes whereas IOP greater than 30 mmHg was observed in 13 (26%) eyes. By day 10, the frequency had decreased to one (2.1 %) eye and three (6.4%) eyes, respectively. The use of antimetabolites did not result in lower IOP or less medication needed for any group at any interval (analysis of variance). CONCLUSION: This modification of a nonvalved glaucoma tube shunt device provides adequate IOP control in the early postoperative period with a low rate of hypotony and surgical complications. If elevation of IOP occurs before suture autolysis, it generally is well controlled by antiglaucoma medications or laser suture lysis. Antimetabolite exposure did not influence early postoperative IOP in this study. PMID- 9855155 TI - Topical verapamil and episcleral venous pressure. AB - OBJECTIVE: To investigate the effect of a single-dose and 2-week administration of topical 0.25% verapamil on episcleral venous pressure (EVP) and intraocular pressure (IOP). DESIGN: A prospective, randomized, double-masked, crossover, and placebo-controlled study. PARTICIPANTS: One eye of each of 20 normal subjects received topical verapamil or placebo. INTERVENTION: The EVP was measured using a pressure chamber episcleral venomanometer. The IOP, EVP, heart rate (HR), and blood pressure (BP) were measured at baseline, 1.5 hours after a single drop, and after 2-week, three-times-a-day topical administration of 0.25% verapamil or placebo to one eye. After a 4-week washout period, the treatment to the study eye was reversed, and measurements of EVP, IOP, HR, and BP were repeated. MAIN OUTCOME MEASURES: The EVP, IOP, HR, and BP were measured. RESULTS: Ninety minutes after a single drop, the EVP in the verapamil-treated eyes was reduced by 4.8% (P = <0.001) and in the untreated fellow eyes by 2.4% (P = 0.032). After 2-week topical administration, the EVP was reduced by 9.5% (P = <0.0001) in the verapamil-treated eyes and by 4.8% (P = 0.001) in the fellow eyes. The EVP was unchanged in the treated and fellow eyes after a single drop or after a 2-week treatment with placebo. Ninety minutes after topical administration of 0.25% verapamil, the IOP decreased by 7.7% (P = <0.0001) in the treated eyes and decreased by 3.6% (P = 0.03) in the fellow eyes. After a 2-week administration of topical verapamil, the IOP decreased by 12% (P = 0.0001) in the treated eyes and by 7.2% (P = 0.0289) in the fellow eyes. The changes of IOP in the placebo treated and fellow eyes were not statistically significant after a single-dose and 2-week administration. After topical administration of verapamil, a contralateral effect on IOP and EVP was observed in the fellow, untreated eye. No systemic effect on HR or BP was detected after a single-dose administration of topical verapamil. There were, however, significant reductions in HR and BP after a 2-week treatment with topical 0.25% verapamil. CONCLUSION: These results indicate that a single drop and a 2-week administration of topical 0.25% verapamil decrease IOP and EVP significantly, with more pronounced reduction after a 2-week treatment than after a single-dose treatment. PMID- 9855156 TI - The rarity of clinically significant rise in intraocular pressure after laser peripheral iridotomy with apraclonidine. AB - OBJECTIVE: To determine the incidence of intraocular pressure (IOP) rise of varying degrees after laser peripheral iridotomy (LPI) in patients with and without glaucoma treated perioperatively with pilocarpine and apraclonidine. DESIGN: A retrospective chart review. PARTICIPANTS: A total of 289 eyes in 179 patients with narrow occludable angles (NOA) (N = 148), open-angle glaucoma or ocular hypertension (OAG) (N = 115), or chronic-angle closure glaucoma (CACG) (N = 26) were reviewed. MAIN OUTCOME MEASURES: The difference between preoperative and postoperative IOP, absolute postoperative IOP, and the need for acute IOP lowering treatment was noted. RESULTS: Only 1.1% (95% confidence interval [CI], 0.03%-5.8%; 1 of 94) of patients and 0.7% (95% CI, 0.02%-3.7%; 1 of 148) of eyes with NOA experienced a rise of more than 10 mmHg 1 to 2 hours after LPI. The incidence of postoperative IOP greater than 25 mmHg and acute postoperative IOP lowering management was 0% (95% CI, 0%-3.8%). Intraocular pressure in 1 of 115 eyes (0.9%, 95% CI, 0.02%-4.7%) with OAG rose more than 10 mmHg, requiring acute treatment. None of the 26 CACG eyes experienced a rise of more than 10 mmHg (95% CI, 0%-13.2%). CONCLUSION: The IOP rise that requires further intervention after LPI with the perioperative use of pilocarpine and apraclonidine is very uncommon. In patients with NOA, routine postiridotomy IOP monitoring may not be required. PMID- 9855157 TI - Intraocular pressure difference in Goldmann applanation tonometry versus Perkins hand-held applanation tonometry in overweight patients. AB - OBJECTIVE: To analyze the increase in intraocular pressure (IOP) caused by anatomic and physiologic factors in overweight patients when using Goldmann applanation tonometry. DESIGN: A prospective cohort study. PARTICIPANTS: Seventy average-weight individuals who had no difficulties with IOP measurements at the slit lamp and 12 obese patients with suspected glaucoma who could position the head at the slit lamp only with great effort participated. INTERVENTION: The authors compared IOP values between slit-lamp-mounted Goldmann applanation tonometry and Perkins hand-held tonometry. MAIN OUTCOME MEASURE: The difference in Goldmann and Perkins IOP measurements was examined. RESULTS: In the group of obese patients, the mean IOP was 20.9+/-2.28 mmHg (mean +/- standard deviation; range, 18-26 mmHg) for the right eye and 21.4+/-3.16 mmHg (range, 16-28 mmHg) for the left eye when determined by Goldmann tonometry and 16.3+/-2.39 mmHg (range, 13-20 mmHg) for the right eye and 16.3+/-2.42 (range, 11-19 mmHg) for the left eye when determined by Perkins tonometry. The mean decrease was 4.5+/-1.3 mmHg (range, 3-7 mmHg) for the right eye and 4.9+/-1.9 mmHg (range, 2-9 mmHg) for the left eye. In the control group, the mean difference between the two types of tonometers for the right eye was 0.34+/-0.69 mmHg and for the left eye was 0.33+/ 0.82 mmHg. Patients who had a falsely elevated IOP on Goldmann tonometry had an average body mass index of 34+/-3.82 (range, 28.5-41.9); most were female (5:1 ratio). CONCLUSION: The authors believe simultaneous breath-holding and thorax compression, with subsequent increase in venous pressure, may be a causative factor for transitory elevations of IOP. Perkins tonometry in obese patients may help avoid a false diagnosis of glaucoma caused by transitory elevations in IOP. PMID- 9855158 TI - Antioxidant nutrient intake and diabetic retinopathy: the San Luis Valley Diabetes Study. AB - OBJECTIVE: Diabetic retinopathy (DR) is a major cause of visual impairment and blindness in adults. Antioxidant nutrients, such as vitamins C and E and beta carotene, may be protective of some eye disorders, such as cataract and age related macular degeneration, but a relationship between these nutrients and DR has yet to be defined. The purpose of this study was to examine the relation between dietary and supplement intakes of vitamins C, E, and beta-carotene and the risk of DR. DESIGN: Both cross-sectional and longitudinal data were collected from participants in the San Luis Valley Diabetes Study, including non-Hispanic white and Hispanic adults in southern Colorado. PARTICIPANTS: A total of 387 participants with type 2 diabetes completed at least 1 complete retinal examination and 24-hour dietary recall (including vitamin supplement use). MAIN OUTCOME MEASURES: Type 2 diabetes was defined according to World Health Organization criteria. DR was assessed by retinal photographs, using the Airlie House criteria to classify DR as none, background, preproliferative, or proliferative. Data for both eyes, from up to three clinic visits per participant, were used for analysis. Ordinal logistic regression analysis was used, taking advantage of multiple clinic visits by individual participants and observations from both eyes, to assess the risk for increased DR severity over time as a function of changes in intake of vitamin C, vitamin E, and beta carotene. Six categories of intake for each nutrient (first to fourth quintiles and ninth and tenth deciles) were considered to ascertain any potential threshold effect. Analyses accounted for age, duration of diabetes, insulin use, ethnicity, glycated hemoglobin, hypertension, gender, and caloric intake. RESULTS: An increase over time in vitamin C intake from the first to ninth deciles was associated with a risk for increased severity of DR (odds ratio = 2.21, P = 0.01), although excess risk was not observed for the tenth decile or the second through fourth quintiles compared to the first quintile. Increased intake of vitamin E was associated with increased severity of DR among those not taking insulin (odds ratios = 2.69, 2.59, 3.33, 5.65, 3.79; P < 0.02, for an increase over time from the first to the second through fourth quintiles and ninth and tenth deciles, respectively). Among those taking insulin, increased intake of beta-carotene was associated with a risk for severity of DR (odds ratio = 3.31, P = 0.003, and 2.99, P = 0.002, respectively, for the ninth and tenth deciles compared to the first quintile). CONCLUSIONS: No protective effect was observed between antioxidant nutrients and DR. Depending on insulin use, there appeared to be a potential for deleterious effects of nutrient antioxidants. Further research is needed to confirm associations of nutrient antioxidant intake and DR. PMID- 9855159 TI - Suprachoroidal hemorrhage: outcome of surgical management according to hemorrhage severity. AB - OBJECTIVE: To report the visual and anatomic outcome after surgical drainage of suprachoroidal hemorrhage according to hemorrhage severity. DESIGN: A retrospective chart review. PARTICIPANTS: Forty-eight consecutive eyes undergoing surgical drainage of a suprachoroidal hemorrhage at The Medical College of Wisconsin were examined. INTERVENTION: Demographic and clinical data were abstracted from patients' medical records. Eyes were classified into four categories of increasing hemorrhage complexity: (1) nonappositional choroidal hemorrhage without vitreous or retinal incarceration in the wound (12 eyes); (2) centrally appositional choroidal hemorrhage without vitreous or retinal incarceration in the wound (17 eyes); (3) choroidal hemorrhage with associated vitreous incarceration in the wound (11 eyes); and (4) choroidal hemorrhage with associated retinal incarceration in the wound (8 eyes). MAIN OUTCOME MEASURES: Visual acuity, rate of persistent hypotony, and incidence of irreparable retinal detachment after surgical drainage for four classes of suprachoroidal hemorrhage were defined. RESULTS: Overall, 11 (23%) of 48 eyes had no light perception (NLP) vision develop, 9 (19%) of 48 eyes had persistent postsurgical hypotony (intraocular pressure < 6), and 21 (64%) of 33 eyes with retinal detachment enjoyed successful retinal reattachment surgery. A definite trend toward an increased rate of NLP vision (P < 0.02), persistent hypotony (P < 0.05), and irreparable retinal detachment (P = 0.11) was observed with increasing suprachoroidal hemorrhage complexity. Eyes with retinal incarceration, compared to eyes without retinal incarceration, had an increased rate of NLP vision (63% vs. 15%; P < 0.01), persistent postsurgical hypotony (50% vs. 13%; P < 0.05), and irreparable retinal detachment (50% vs. 20%; P = 0.07). CONCLUSIONS: Eyes requiring surgical drainage of a suprachoroidal hemorrhage have a guarded prognosis, with a poorer outcome associated with increasing hemorrhage complexity. A classification system incorporating choroidal apposition, and vitreous and retinal incarceration in the wound, provides a format for reporting and assessing the efficacy of management strategies in this condition. PMID- 9855160 TI - Factors associated with elevated rates of adverse outcomes after cyclodestructive procedures versus drainage device procedures. AB - OBJECTIVE: To identify factors associated with an increased risk of adverse outcomes after cyclodestructive or drainage device procedures. DESIGN: Retrospective, cohort analysis. PARTICIPANTS: A total of 5570 Medicare patients who were older than 65 years of age and who underwent cyclodestructive or drainage device procedures in 1994 participated. INTERVENTION: The authors identified cyclodestructive and drainage device procedures from claims to the Health Care Finance Administration (HCFA) by International Classification of Diseases (ICD-9) procedure codes, Current Procedural Terminology procedure codes, and HCFA Common Procedural Classification System codes. The authors analyzed adverse outcome rates using hierarchical logistic regression. Race, age group, gender, length of observed follow-up, state in which surgery took place, ocular procedures performed before and at the same time as the index surgery, and ocular diagnosis were included as covariates in the model. MAIN OUTCOME MEASURES: The authors defined an adverse outcome as the occurrence after the index surgery of at least one of the following: repeat cyclodestructive or drainage device procedure, retinal hole-tear repair, retinal detachment repair, surgery for endophthalmitis, vitrectomy, enucleation, evisceration, surgery for ocular hypotony, and/or extrusion or revision of drainage device. Adverse outcomes were also defined without the inclusion of repeat cyclodestructive or drainage device procedures. RESULTS: When repeat cyclodestructive or drainage device procedures were not included in the definition of an adverse outcome, eyes with a drainage device procedure were 3.8 times more likely to have an adverse outcome than eyes with a cyclodestructive procedure (odds ratio [OR], 3.8; 95% confidence interval [CI], 3.07, 4.67). Subjects with concurrent corneal transplant had increased odds of an adverse outcome compared to subjects without a concurrent corneal transplant (OR, 2.00; 95% CI, 1.27, 3.15). When the definition of an adverse outcome included repeat cyclodestructive or drainage device procedures, the odds of an adverse outcome were similar for both cyclodestructive and drainage device procedures (OR, 0.94; 95% CI, 0.79, 1.13). CONCLUSIONS: Cyclodestructive procedures need to be repeated more frequently than drainage device procedures. However, if the patient has a drainage device procedure, then that patient is more likely to have other types of adverse ophthalmic events than if he or she had a cyclodestructive procedure. Because the average follow-up of subjects in this study is 5 months (range, 0-12 months), outcomes that might take longer to manifest themselves would be excluded from this study. PMID- 9855161 TI - Primary vitrectomy for rhegmatogenous retinal detachment associated with choroidal detachment. AB - OBJECTIVE: This study aimed to report the success rate of primary vitrectomy, scleral buckling, and oral steroids in eyes with combined rhegmatogenous retinal detachment (RRD) and choroidal detachment (CD) and to compare these results with those reported in the literature for similar cases managed by scleral buckling alone. DESIGN: Consecutive case series. PARTICIPANTS: Twenty-one eyes of 21 consecutive patients with primary RRD associated with CD were examined. INTERVENTION: All patients were treated with systemic corticosteroids before surgery. Pars plana vitrectomy with scleral buckling or encircling or both were performed. MAIN OUTCOME MEASURES: Retinal reattachment rate and resolution of choroidal detachment were measured. RESULTS: At mean follow-up of 11.4 months, retinal reattachment was attained in 19 eyes (90.5%) compared to less than 53% reported in the literature for scleral buckling alone. With preoperative oral steroids, choroidal detachment regressed completely in 13 eyes (61.9%). In the remaining eyes with persistent choroidal detachment, suprachoroidal fluid was drained during vitrectomy. CONCLUSION: Aggressive treatment with oral steroids followed by pars plana vitrectomy and scleral buckling or encircling is recommended instead of scleral buckling alone in the management of combined primary rhegmatogenous retinal detachment and choroidal detachment. PMID- 9855162 TI - X-linked retinitis pigmentosa in two families with a missense mutation in the RPGR gene and putative change of glycine to valine at codon 60. AB - OBJECTIVE: This study describes the ophthalmic findings in two unrelated white families with X-linked retinitis pigmentosa (XLRP) caused by a missense mutation in the retinitis pigmentosa GTPase regulator (RPGR) gene. DESIGN: Genetic screening and clinical correlation. PARTICIPANTS: Thirty-six families with XLRP seen by the authors were screened for a possible mutation in the RPGR gene to identify three affected hemizygotes with retinitis pigmentosa and four heterozygote carriers in one family and one hemizygote and one carrier in a second family. INTERVENTION: All nine patients underwent a routine ocular examination, including slit-lamp biomicroscopy and a dilated fundus examination. Goldmann visual field kinetic perimetry, static threshold perimetry, and electroretinography also were obtained. The DNA screening was performed on the three affected male patients and four obligate carriers examined from one family and the two examined patients, plus an additional male and obligate carrier, from the second family to determine the presence of any causative mutation in the RPGR gene. MAIN OUTCOME MEASURES: Findings on fundus examination, static threshold and kinetic perimetry, and electroretinography testing were the main outcome measures. RESULTS: A G-->T nucleotide change at position 238 in exon 3 of the RPGR gene resulting in a putative substitute of Gly-->Val at codon 60 was shown to segregate with RP in affected males and the carrier state in female heterozygotes in these two families. The ophthalmologic findings in hemizygotes as well as the carriers in this family were within the spectrum of findings characteristically noted in XLRP families. A tapetal-like reflex was not observed in any of the five female carriers. Psychophysical and electrophysiologic testing on the carriers indicated that cone and rod functions were impaired equivalently. When present in the carriers, visual field restriction was most apparent in, or limited to, the superotemporal quadrant, which corresponded to the retinal pigmentary changes that tended to occur in the inferonasal retina. CONCLUSIONS: A mutation in exon 3 of the RPGR gene, which would result in a putative glycine to valine substitution at codon 60, is associated with a severe clinical phenotype in male patients and a patchy retinopathy without a tapetal-like reflex in carrier females. In these families, heterozygote carriers showed equivalent impairment of their cone and rod function. PMID- 9855163 TI - Laser treatment of feeder vessels in subfoveal choroidal neovascular membranes: a revisitation using dynamic indocyanine green angiography. AB - OBJECTIVE: This study aimed to determine whether the indocyanine green angiography (ICGA)-guided laser treatment of feeder vessels (FVs) may be useful in the management of the subfoveal choroidal neovascular membranes (CNVM) in patients with age-related macular degeneration (ARMD). DESIGN: Noncomparative case series. PARTICIPANTS: The authors considered a series of 15 patients with subfoveal CNVM in whom feeder vessels could be clearly detected by means of dynamic ICGA but not necessarily with fluorescein angiography (FA). On the basis of the indications of the pilot study, the authors also studied a second series of 16 patients with FVs smaller than 85 microm. INTERVENTION: Treatment of FV using argon green laser was performed. The ICGA was performed immediately after treatment, after 2, 7, 30 days, and then every 3 months, to assess FV closure. If an FV appeared to be still patent, it was immediately retreated and the follow-up was started again. The follow-up time ranged from 23 to 34 months for the pilot study and from 4 to 12 months for the second series. MAIN OUTCOME MEASURES: The obliteration of the membrane and change in visual acuity from baseline were measured. The effect on the treatment of the number and width of the FVs, and the size and location of the membrane, also was evaluated. RESULTS: In the pilot study, the CNVM was obliterated after the first treatment in only one patient, five patients needed more than one treatment, and obliteration failed in nine patients (40% success rate). The rate of success was affected by the width and number of the FVs. The success rate in the second series of 16 patients was higher (75%). CONCLUSIONS: The success of the laser treatment of FVs depends on their width, length, and number. Dynamic ICGA, which detects smaller FVs and makes it possible to control the laser effect and initiate immediate retreatment in the case of incomplete FV closure, should be considered mandatory for this type of treatment; a comparable success rate would have been unlikely using the other currently available methods of treating subfoveal CNVMs. PMID- 9855164 TI - Abnormal cone synapses in human cone-rod dystrophy. AB - OBJECTIVE: Little is known of the cytopathology of photoreceptors in human inherited retinal dystrophies that initially affect the central retina, including the macula. The current study sought to determine the cytologic features of dysfunctional cone and rod photoreceptors, as well as the pattern of degeneration of the cells in representative cases of central retinal dystrophy. STUDY DESIGN: Comparative human tissue study. MATERIALS: Four human donor eyes with the following forms of central retinal dystrophy: cone-rod dystrophy (CRD), central areolar choroidal dystrophy, Bardet-Biedl syndrome, and cone dystrophy-cerebellar ataxia. The cytologic features of retinal photoreceptors in these eyes were compared with those in an eye with retinitis pigmentosa and six normal human eyes. METHODS AND OUTCOME MEASURES: Immunocytochemistry and electron microscopy were used to evaluate the retinal histopathology in the donor eyes. RESULTS: Cone numbers were decreased in the case of CRD, particularly in the central and far peripheral retina, and both cone and rod outer segments were slightly shortened. Occasional degenerate cones had dense cytoplasm and pyknotic nuclei dislocated sclerad to the external-limiting membrane. The most prominent alteration in this retina was marked enlargement and distortion of the cone photoreceptor pedicles, which contained reduced numbers of synaptic vesicles. The retina with central areolar choroidal dystrophy contained a few cones with similarly abnormal synapses. However, comparable cone synapse abnormalities were not observed in the cases of Bardet-Biedl syndrome, cone dystrophy-cerebellar ataxia, retinitis pigmentosa, or in the normal retinas. CONCLUSIONS: The functional consequences of the cone synapse abnormalities in CRD are not known but may correlate with the electroretinographic abnormalities documented in some cases of CRD. To our knowledge, comparable synapse changes have not been noted in either rods or cones in other forms of retinal dystrophy, including retinitis pigmentosa, suggesting that different cytopathologic mechanisms may be involved. PMID- 9855166 TI - Screening for ophthalmic disease in older subjects using visual acuity and contrast sensitivity. AB - OBJECTIVE: Despite early interest in contrast sensitivity as a screening test for ophthalmic disease, most published opinion suggests that there is no benefit over conventional measurement of visual acuity. Taking a primary care perspective of screening, the authors evaluated the ability to discriminate those with any diagnosed ophthalmic disease in a large sample representative of the general population. DESIGN: Retrospective analysis of a clinical, cross-sectional survey. Snellen visual acuity, contrast sensitivity (Arden plates, American Optical contrast sensitivity test), and ophthalmic diagnosis were reported previously. PARTICIPANTS: A sample of 3283 subjects, all aged at least 50 years, were selected randomly from residents of a health district in Sydney, Australia. Ophthalmologic diagnosis (ophthalmic disease presence/absence) had been confirmed for 2522 of these subjects. MAIN OUTCOME MEASURES: Signal detection techniques (the receiver-operating characteristics function [ROC], quality ROC [QROC], and weighted kappa coefficient of association [kappa(r)]) were used to evaluate test discriminability. RESULTS: Because analyses of right and left eyes were almost identical, only right eye results are presented. Advantages of kappa(r) over ROC were shown. Discrimination of those with diagnosed ophthalmic disease from those without ophthalmic disease was best with Arden plate 7 (kappa0.5 = 0.93) and was better than distance Snellen visual acuity (kappa0.5 = 0.59). Arden plate 7 (6.4 cyc/deg) correctly assigned 96% of subjects at its optimal pass-fail criterion. CONCLUSIONS: In the primary care setting, a person older than 50 years of age with reduced contrast sensitivity, as determined by Arden plate 7, requires extra care in subsequent examinations because this person is likely to have an ophthalmic disease. PMID- 9855165 TI - The role of weight loss and acetazolamide in the treatment of idiopathic intracranial hypertension (pseudotumor cerebri) AB - OBJECTIVE: To determine the weight loss associated with resolution of papilledema from idiopathic intracranial hypertension (IIH). DESIGN: A retrospective study. PARTICIPANTS: Fifteen consecutive female patients with IIH associated with obesity were studied. INTERVENTION: Patients underwent weight loss and treatment with acetazolamide during a 24-week period. MAIN OUTCOME MEASURES: The severity of papilledema was graded: absent (grade 0), mild (grade 1), moderate (grade 2), and marked (grade 3), based on a predetermined grading system ("gold standard") using stereoscopic photographs and the Frisen classification. RESULTS: The 15 patients, with mean age of 31.3+/-8.8 years, had a mean weight of 110.5+/-28.7 kg and mean body mass index of 40.7+/-13.0 kg/m2. Eleven (73.3%) patients had improved papilledema during the 24-week study period, of which 10 (66.7%) had complete resolution of papilledema within a median time of 8.5 weeks. An average of 3.3% weight loss (+/-0.5% standard error of the mean) was observed among patients having a one-grade change in papilledema. Weight loss of 6.2%+/-0.6% standard error of the mean was associated with a three-grade change in papilledema (i.e., complete resolution of marked papilledema). Nine of the ten patients with complete resolution of papilledema also took acetazolamide. However, none (26.7%) of the four patients without weight loss had improvement in papilledema despite similar treatment with acetazolamide. CONCLUSIONS: Approximately 6% weight loss was associated with resolution of marked papilledema in these authors' patients. The benefit of acetazolamide in IIH is questioned since weight loss, rather than acetazolamide, appeared to have been the catalyst for reducing the severity of papilledema. PMID- 9855167 TI - Elevated tear interleukin-6 levels in patients with Sjogren syndrome. AB - OBJECTIVE: The purpose of the study was to investigate interleukin-6 (IL-6) levels in the tear fluid and sera of patients with Sjogren syndrome (SS). PARTICIPANTS: Twelve patients with primary SS and 12 normal control subjects participated. INTERVENTION: Tear fluid and sera were obtained from the study and the control groups. Evaluation of tear fluid and sera IL-6 levels was done by using a quantitative enzyme-linked immunosorbent assay kit. All assays were carried out blindly with respect to diagnosis. MAIN OUTCOME MEASURES: Tear fluid IL-6 levels were measured. RESULTS: The mean concentration (+/- standard error) of IL-6 in the tears of patients with SS was elevated significantly compared to that of normal control subjects (88.6+/-16.2 vs. 42.1+/-10.6 pg/ml; P < 0.05). No significant differences were noted in the serum IL-6 levels between the two groups. A significant correlation (r = 0.742, P = 0.006) was found between tear fluid IL-6 levels and the focus score of lip biopsy specimens in patients with SS. CONCLUSION: Tear fluid IL-6 levels may serve as an important marker for tear gland involvement in SS. PMID- 9855168 TI - Soft lens extended wear affects epithelial barrier function. AB - OBJECTIVE: To explore the impact of eye closure and soft contact lens extended wear (SCLEW) on epithelial permeability to fluorescein (Pdc). DESIGN: A prospective cohort study. PARTICIPANTS: Thirty-one noncontact lens (CL) wearers participated. INTERVENTION: The effects of eye closure on Pdc were evaluated by comparing morning (AM) and afternoon (PM) measurements on non-CL subjects. The effects of SCLEW on Pdc were determined by measuring Pdc before beginning SCLEW and then after 2, 4, and 12 weeks of SCLEW. MAIN OUTCOME MEASURES: Pdc measured in the morning versus the afternoon and before versus after SCLEW was examined. RESULTS: Analyses of Pdc were done using the natural logarithm (ln). The mean +/- standard error (SE) ln(Pdc) measured in the AM versus PM on 16 non-CL wearers did not differ significantly (-2.56+/-0.16 vs. -2.69+/-0.15, respectively; P = 0.46). In contrast, the mean +/- SE ln(Pdc) in 15 subjects after 2 (-1.73+/-0.183, P < 0.001), 4 (- 1.59+/-0.188, P < 0.001), and 12 weeks (-1.99+/-0.206, P = 0.02) of SCLEW was substantially greater than that measured before lens wear (-2.42+/ 0.159 ln(nm/sec)). Interestingly, the mean+/-SE ln(Pdc) measured in the afternoon on seven subjects after 13 weeks of SCLEW was similar to their average baseline ln(Pdc) (-2.62+/-0.27 vs. -2.52+/-0.243, respectively; P = 0.54). Further analysis showed that Pdc was highest in the morning and decreased approximately 12.5%/hour (P < 0.001) during the day. CONCLUSIONS: Sleeping without CLs does not appear to alter Pdc; however, 2 weeks of SCLEW caused a 99% increase in permeability without observable changes by slit-lamp examination. Increases in Pdc appear greatest in the morning after SCLEW and then decrease exponentially during the day. Whether changes in Pdc will predict CL-associated keratopathy needs further study. PMID- 9855169 TI - Nonsteroidal anti-inflammatory drug-related gastrointestinal toxicity: definitions and epidemiology. AB - Nonsteroidal anti-inflammatory drug (NSAID)-associated gastrointestinal (GI) toxicity is a broad topic encompassing symptoms as well as severe GI complications. GI bleeding and perforation are the 2 overlapping components that account for the majority of deaths and disability associated with these drugs. Abnormal gastric endoscopic profiles as well as symptoms such as heartburn, pain, and dyspepsia are common in NSAID users, but no correlation has been found between these factors and the occurrence of the more severe complications; therefore, neither symptoms nor endoscopic observations can necessarily be considered reliable predictors of future outcomes. Confounding factors can increase the risk of complications, and specific NSAIDs vary in the magnitude and type of risk attending their use. Recent studies have focused on the contribution of nonprescription NSAIDs to total complications, and combined with evidence suggesting that the risk is greatest during the first month of NSAID use, it is apparent that NSAID toxicity is an acute as well as a chronic problem. PMID- 9855170 TI - Pathogenesis of nonsteroidal anti-inflammatory drug-related upper gastrointestinal toxicity. AB - The mechanisms involved in the pathogenesis of ulcers associated with the use of nonsteroidal anti-inflammatory drugs (NSAIDs) have not been fully elucidated. Although studies using acute mucosal injury as a surrogate for clinically relevant outcomes have provided useful information, in practice, acute mucosal injury does not necessarily provide a reliable predictor of clinical ulcers or complications. Several factors that increase the risk of NSAID-associated gastroenteropathy have been identified, and there are data to support or provide speculation for other physiologic factors that might predispose specific subsets of patients to increased mucosal injury. Since clinically significant events occur less frequently than does ulceration, it is the latter determinant, i.e., the identification of the traits that distinguish patients who develop serious NSAID-associated gastrointestinal (GI) toxicity from those who can tolerate these drugs, that may provide the clues necessary to understand the events underlying ulcer pathogenesis. PMID- 9855171 TI - Nonsteroidal anti-inflammatory drug-associated toxicity of the liver, lower gastrointestinal tract, and esophagus. AB - Although upper gastrointestinal (GI) adverse events are the most common consequences of nonsteroidal anti-inflammatory drug (NSAID) use, there are other GI side effects that can contribute to the morbidity and mortality associated with these drugs. NSAID-associated toxicity of the large and small bowel is increasingly recognized in clinical practice, as enteroscopic procedures become more frequently used. This lower GI toxicity may have several different manifestations: ulcerations, strictures, colitis, or exacerbation of inflammatory bowel disease. Hepatic injury, most likely due to an idiosyncratic reaction resulting from an immunologic response or altered metabolic pathways, is another sequela of NSAID use that is usually reversible. Although hepatotoxicity is listed as a class warning for NSAIDs, aspirin, diclofenac, and sulindac are most commonly associated with this problem. Surveillance for hepatic injury is not always reliable, and the low frequency of both hepatic and lower GI toxicity in NSAID users renders these events difficult to characterize. An increase in awareness, surveillance, and reporting of these events can lead to a better understanding of the risk factors and etiology associated with NSAID toxicity. PMID- 9855172 TI - The relation between Helicobacter pylori and nonsteroidal anti-inflammatory drugs. AB - Both infection with Helicobacter pylori and use of nonsteroidal anti-inflammatory drugs (NSAIDs) can result in gastritis and ulcers. H. pylori has been identified as a major etiologic factor in the development of peptic ulcer disease; however, its relationship to NSAID-associated toxicity is less well characterized. Several studies have suggested that NSAID use does not increase susceptibility to H. pylori, and the converse has also been suggested, namely, that H. pylori does not exacerbate NSAID-associated injury. H. pylori itself may stimulate production of gastric prostaglandins, which may have a role in ulcer healing. More carefully controlled studies may be better able to elucidate the individual and synergistic mechanisms involved in ulceration induced by H. pylori and NSAIDs. Recent studies have suggested that elimination of H. pylori before NSAID treatment decreases ulcer occurrence. Therefore, at this time, eradication of H. pylori should be considered only in certain high-risk patients, i.e., those with a history of gastroduodenal ulcers. PMID- 9855173 TI - Role of endoscopy in nonsteroidal anti-inflammatory drug clinical trials. AB - Endoscopy is a useful tool that can be used to determine the acute or chronic gastrointestinal (GI) toxicity of nonsteroidal anti-inflammatory drugs (NSAIDs) and to confirm outcomes in clinical trials. However, since evaluations of endoscopic injuries are to some extent subjective (e.g., the endoscopic distinction between an erosion and an ulcer), such injuries must be clearly and correctly defined before the data can be analyzed and considered meaningful. Definitions of injury, endpoints, and protocol design must be consistent with the intent of the study. This becomes evident in drawing distinctions between acute injury, which may resolve, and chronic injury that occurs over a longer period of time. The intent of the study itself should be clearly defined and based on realistic and realizable goals. Only when these criteria are met, and preferably standardized, can clinically relevant studies be performed and compared. PMID- 9855174 TI - Agents used in the prevention and treatment of nonsteroidal anti-inflammatory drug-associated symptoms and ulcers. AB - Coprescription of gastroprotective agents is a common practice for patients requiring nonsteroidal anti-inflammatory drug (NSAID) therapy, although there is limited evidence that some of these agents are effective. The prostaglandin analog misoprostol and the proton pump inhibitor omeprazole appear to be efficacious in decreasing NSAID-associated ulcers. Misoprostol has also been shown to decrease NSAID-associated gastrointestinal (GI) complications by 40% compared with placebo. Despite the efficacy of these drugs, their effectiveness in the general population has not yet been adequately determined with respect to reduction of symptoms and improvement in patient quality of life. Sucralfate and bismuth appear to be largely ineffective, and histamine receptor antagonists, when given at traditional ulcer-healing doses, decrease symptoms and duodenal ulcers only. The issue of outcomes research, therefore, needs to be more fully incorporated into any analysis of the effectiveness or cost-effectiveness of the widespread clinical use of such gastroprotective drugs. PMID- 9855175 TI - Safety profiles of leading nonsteroidal anti-inflammatory drugs. AB - Recently introduced nonsteroidal anti-inflammatory drugs (NSAIDs) have capitalized on new formulations or unique physical and pharmacologic properties in an attempt to provide a greater margin of gastrointestinal (GI) safety. The use of enteric coatings and nonoral or pro-drug formulations have not necessarily provided the expected safety, but other properties have been identified that appear to be more promising. However, as demonstrated by oxaprozin, considered to be one of the least safe NSAIDs but one of the leading drugs on the US market, success may not be dependent on safety. In contrast, the improved tolerability of 2 other new NSAIDs, nabumetone and etodolac, has been established in clinical trials and limited postmarketing surveillance. This improved tolerability is probably associated with several pharmacologic properties that have been suggested to contribute to GI safety: (1) nonacidic pro-drug formulation; (2) lack of enterohepatic recirculation; (3) a short plasma half-life; and (4) preferential inhibition of cyclo-oxygenase-2 (COX-2). Although these factors may not improve efficacy, their incorporation into the design of drugs suggests that safer NSAIDs may be a clinical reality. However, the safety and clinical value of any new drug for the general population will be validated only through extensive postmarketing surveillance. PMID- 9855176 TI - Future trends in the development of safer nonsteroidal anti-inflammatory drugs. AB - Gastrointestinal (GI) adverse events, ranging from mild to life-threatening, are well-recognized sequelae to nonsteroidal anti-inflammatory drug (NSAID) use. Recent improvements in our knowledge of the mechanisms responsible for NSAID associated gastropathy have enabled several experimental approaches to decreasing the risk of these events. Whereas such strategies as preassociation of NSAIDs to zwitterionic phospholipids to prevent NSAID-mucosal interactions and concomitant administration of trefoil peptides to stimulate mucosal defense pathways represent novel approaches, their clinical feasibility remains to be determined. Other strategies that appear more immediately promising in the reduction of NSAID associated GI toxicity are the coupling of NSAIDs to nitric oxide (NO)-releasing compounds and the introduction of NSAIDs that are preferential or specific for cyclo-oxygenase-2 (COX-2), the isoform implicated in the inflammatory response. Clinical trials of several specific COX-2 inhibitors, as well as European clinical data for a preferential COX-2 inhibitor, meloxicam, suggest that COX-2 inhibitors provide an advantage over standard NSAIDs in terms of GI tolerability. However, as recent observations have implicated COX-2 as an integral component in the maintenance of physiologic homeostasis, careful scrutiny of both the beneficial and the deleterious effects of the selective COX-2 inhibitors is requisite before their approval and widespread use. Furthermore, based on the physiologic importance of COX-2, the preferential inhibitors may ultimately prove to represent the optimal compromise for the treatment of various arthritides. PMID- 9855177 TI - Heparin-induced thrombocytopenia. PMID- 9855178 TI - Pathophysiology and laboratory testing of heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) type II is an immune-mediated reaction that generally occurs 5 to 14 days after initiation of heparin therapy. It is characterized by a severe decline in platelet count (by >50%) in association with a new thromboembolic complication. Type II HIT is mediated by antibodies, usually of the IgG class. The antibodies cause platelet activation in the presence of heparin or other polysulfated saccharides. The antigen in type II HIT frequently is a complex of platelet factor 4 (PF4) and heparin. In addition to stimulating platelet activation, type II HIT antibodies bind to PF4/heparin complexes on the surface of endothelial cells, leading to activation of the cells. Concomitant activation of platelets and endothelial cells together with enhanced thrombin generation are likely causes for the thromboembolic events. The percentage of patients receiving heparin who develop type II HIT antibodies is higher than the percentage of patients who develop clinical symptoms characteristic of type II HIT. Therefore, it is currently not indicated to screen asymptomatic patients for the development of HIT antibodies. If clinical symptoms of HIT appear, the clinical diagnosis should be confirmed, when possible, by a laboratory test for HIT antibodies. Assays for detection of HIT antibodies are either functional assays, which measure heparin-dependent platelet activation, or antigen assays based on the enzyme-linked immunosorbent assay (ELISA) technique. A reasonable approach to applying these laboratory tests is to use one assay technique as a screening test and reserve the other for testing samples that had negative screening results when the diagnosis of HIT is strongly suspected. PMID- 9855179 TI - Clinical presentation of heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is an immune-mediated adverse effect of heparin that is important because of its relatively high frequency and its strong association with paradoxic venous and arterial thrombosis. As confirmatory laboratory assays are not always immediately available, physicians usually must make initial diagnostic and treatment decisions based on the clinical presentation alone. Three characteristic features of HIT can be helpful in distinguishing it from other causes of thrombocytopenia: (1) timing of the onset of thrombocytopenia, namely, a platelet count decrease that begins between days 5 and 8 (inclusive) of beginning heparin treatment; (2) mild to moderate severity of the thrombocytopenia, with platelet counts only rarely less than 15 x 10(9)/L; and (3) the development of large-vessel venous or arterial thrombosis in association with thrombocytopenia, or any of a number of unusual characteristic sequelae of HIT (warfarin-associated venous limb gangrene, bilateral adrenal hemorrhagic infarction, heparin-induced skin lesions, or acute systemic reactions following intravenous heparin bolus). In contrast to other drug-induced immune thrombocytopenia syndromes, HIT rarely is associated with bleeding. HIT is relatively common, occurring in as many as 3% of patients who receive unfractionated (UF) heparin for 2 weeks. Between 30% and 75% of patients with HIT develop thrombosis; thus, about 1% of patients who receive a course of heparin develop HIT-associated thrombosis. The observation that HIT is less likely to occur with low-molecular-weight heparin (LMWH) suggests that HIT ultimately may be preventable. PMID- 9855180 TI - Limitations of conventional treatment options for heparin-induced thrombocytopenia. AB - Thrombosis is a common and potentially serious complication of immune-mediated heparin-induced thrombocytopenia (HIT). Discontinuation of heparin is a simple and important maneuver in patients with suspected HIT. Unfortunately, thrombosis often occurs even in those patients in whom heparin was discontinued because of thrombocytopenia alone ("isolated" HIT). It therefore is reasonable to consider prophylactic anticoagulation with an alternate anticoagulant in patients with suspected HIT, especially if their initial indication for anticoagulation persists. For patients with thrombosis complicating HIT, conventional treatment options often have important limitations. Warfarin has a slow onset of action, and its use in patients with acute HIT and deep venous thrombosis has been associated with the devastating syndrome of venous limb gangrene. Ancrod, a defibrinogenating snake venom with thrombin-like activity, has also been used to treat HIT. However, this agent does not inhibit thrombin generation in HIT, which could explain why some patients who have been treated with this agent have developed certain adverse clinical events, such as warfarin-associated venous limb gangrene. The use of low-molecular-weight heparin (LMWH) to treat patients with HIT is limited by their high rate (up to 100%) of in vitro cross-reactivity with HIT sera, and the relatively frequent occurrence of new or recurrent thrombocytopenia or thrombosis during treatment of HIT with this class of agents. In contrast, the mixture of anticoagulant glycosamingoglycans known as danaparoid sodium has a much lower frequency of in vitro cross-reactivity with HIT sera (10% to 40%, depending upon the sensitivity of the assay). Moreover, clinically significant cross-reactivity during treatment with danaparoid appears to be uncommon, even in patients in whom in vitro cross-reactivity is demonstrable. PMID- 9855181 TI - New treatment options for heparin-induced thrombocytopenia. AB - A rapidly acting anticoagulant that can either inhibit thrombin generation or inhibit thrombin itself is the optimum therapy for acute thrombosis associated with heparin-induced thrombocytopenia (HIT). In this review, the newer treatment approaches that fulfill this requirement are discussed. These newer treatments include hirudin and argatroban, direct thrombin inhibitors, and danaparoid, which inhibits thrombin generation. Preliminary outcome results from the extensive compassionate-use program for danaparoid in HIT and from a recently completed randomized clinical trial that compared danaparoid with dextran in patients with HIT are provided. Based on these data, danaparoid appears to be a useful and safe replacement for heparin in patients who develop HIT. PMID- 9855182 TI - Microcapsules of alginate-chitosan--I. A quantitative study of the interaction between alginate and chitosan. AB - The binding of chitosan to alginate beads was studied quantitatively by using radioactive labelled fractions of chitosan. The alginate-chitosan capsules were made either by dropping a solution of sodium alginate into a solution containing chitosan or by incubating calcium alginate beads in a solution of chitosan. The first procedure yielded a binding of 0.015 microg chitosan per mm2 of capsule surface, while the latter procedure yielded over 2 microg mm(-2). The maximum obtained weight ratio of chitosan to alginate in a microcapsule after 24 h was 0.40. The binding of chitosan was markedly increased by reducing the number average molecular weight of chitosan below 20000 Da and by increasing the porosity of the alginate gel. The porosity was increased by producing homogeneous gels, and by adding calcium chloride to the chitosan solution during the membrane forming stage. The effect of calcium ions on the porosity of the gel was studied by experiments involving release of blue dextran from calcium alginate beads. The binding of chitosan was also found to increase with decreasing fraction of N acetylations, FA, on chitosan in the range of FA = 0.3 to FA = 0, and with increasing pH in the range from pH 4 to 6. Capsules with a diameter of 500 microm had a higher weight ratio of chitosan to alginate after 24 h of binding than the capsules with the larger diameter of 1500 microm. PMID- 9855183 TI - Biodegradable polyesters for controlled release of trypanocidal drugs: in vitro and in vivo studies. AB - Copolymers of epsilon-caprolactone and L-lactide P(CL-LLA), epsilon-caprolactone and D,L-lactide P(CL-DLLA) and epsilon-caprolactone and trimethylene carbonate P(CL-TMC) were synthesized. The composition of comonomers and their sequence lengths were determined by means of 1H and 13C NMR measurements. The effect of the comonomer on the thermal properties was investigated by differential scanning calorimetry (DSC) analysis. The in vitro degradation of the rods obtained by melt extrusion of the synthesized copolymers and the commercial homopolymers poly(epsilon-caprolactone) P(CL) and poly(D,L-lactide) P(DLLA) was carried out in phosphate buffer (PB) pH 7.4 at 37 degrees C. The rate of degradation depends on comonomers and polymer composition. The in vitro release of the selected drugs, isometamidium chloride (IMM) and ethidium bromide (EtBr), from such devices was carried out under the same conditions as used for the in vitro degradation. The release experiments show that the release of IMM is faster than for EtBr. During the first stage, for IMM the release is governed by osmotic pressure whereas for EtBr the release is mainly diffusion-controlled. The in vitro release of these drugs is governed by polymer matrix degradation at the later stage of the release process. Comparative in vitro release study from the different polymers showed that the release depends mainly on the physical properties of the polymer. The in vivo experiments carried out in the field on cattle and in the laboratory on rabbits using the classical treatment (intramuscular injection) and the sustained release devices (SRD) subcutaneously implanted, showed that the prophylactic period is significantly enhanced in the case of SRD as compared to intramuscular injection. The comparative efficacy of SRD containing IMM and EtBr evaluated in the case of rabbits showed that, the SRD (IMM) prophylactic period is much longer than for SRD (EtBr). PMID- 9855184 TI - Viscous property and osteogenesis induction of hydroxyapatite thermal decomposition product mixed with gelatin implanted into rabbit femurs. AB - The viscosity of hydroxyapatite thermal decomposition product mixed with gelatin (HATDP-G) and the osteogenesis induced by this mixture were investigated. To determine the viscosity of HATDP-G, we measured the average diameter of a slumped mass of HATDP-G (0.5 ml) according to the American Dental Association Specification No. 8, with some modifications, and compared it with that of an equal mass of HATDP and of polymethylmethacrylate (PMMA) bone cement. The mean diameter of 10 samples each of HATDP-G, HATDP, and PMMA bone cement was 26.8+/ 0.9 mm, 19.6+/-0.2 mm and 29.2+/-0.3 mm, respectively, 2 min after the mixing procedure. HATDP-G was injected into the bone marrow of the right femurs of 25 rabbits. As the control, gelatin with saline solution was injected into the left femurs. New bone formation was observed in all 20 femurs from three weeks after injection. No new bone formation was observed in the control femurs. The affinity index, a parameter of osteogenesis, was 19.8+/-6.1%, 27.5+/-2.6%, 52.9+/-9.5%, and 76.3+/-1.9% at 3, 6, 12 and 24 weeks after the injection, respectively. Significant increases in the affinity index were found between weeks 6 and 12, and weeks 12 and 24 (p < 0.05). Our findings indicated that HATDP-G might be successfully used as a bioactive bone cement. PMID- 9855185 TI - Growth and differentiation of human bone marrow osteoprogenitors on novel calcium phosphate cements. AB - Materials that augment bone cell proliferation and osteogenic activity have important therapeutic implications for bone regeneration and for use in skeletal reconstruction and joint replacement. We have studied the growth and interactions of human bone marrow cells on a variety of new cement composites in vitro. These cement materials are composed of calcium-deficient hydroxyapatites, carbonated apatite and amorphous calcium phosphate. Cell proliferation was significantly reduced and cell differentiation increased in the presence of these cements compared with cells cultured on tissue culture plastic. Alkaline phosphatase, one of the markers of the osteoblast phenotype, was dramatically stimulated by 3 of the 4 cements examined between day 4 and day 10, above levels observed following culture of human osteoblasts on plastic alone. Photomicroscopic examination demonstrated growth and close integration of bone marrow cells and 3 of the composites. Longer term marrow cultures (15 day) on the cements confirmed the stimulation of cell differentiation over proliferation. From these studies, enhanced osteoblastic differentiation was observed on a 70% carbonated apatite, which has a composition similar to bone mineral, whereas, cell toxicity was observed on cells grown on amorphous calcium phosphate. This in vitro culture system demonstrates the use of human bone marrow cells for the potential evaluation of new biomaterials and the development of a novel carbonated apatite that may be of potential use in orthopaedic implants. PMID- 9855186 TI - Biodegradation of ornithine-containing polylysine hydrogels. AB - The degradation of the cross-linked cationic poly(amino acid)-glutaraldehyde (GA) hydrogels by two kinds of proteolytic enzymes, trypsin and Aspergillus Protease Type XXIII, and by seven species of soil filamentous fungi has been investigated using homo- and copolypeptides of lysine (Lys) and ornithine (Orn). Trypsin degraded the hydrogels prepared from poly(Lys) and copoly(Lys Orn)s but not poly(Orn), while Aspergillus protease degraded all of them. Degradation time of hydrogels by the two proteases became longer with increasing Orn content in the gel. Seven species of soil filamentous fungi were cultured with hydrogels on Czapeck medium to evaluate the degree of microbial degradation of the hydrogels, and the three species of the fungi, Aspergillus oryzae, Penicillium citrinum and Curvularia sp., were grown in culture with an accompanying degradation of the gel matrix, while the other four species, Mucor sp., Rhizopus sp., Cladosporium sp., and Trichoderma sp., were not. The degree of degradation of gel matrix with growth of the three fungi became lower with increasing Orn content in the gel matrix. The results might offer some clues to the applications for the controlled biodegradation of cationic poly(amino acid) hydrogel by introduction of Orn, suggesting that unnatural amino acid resists hydrolysis by proteases or microorganisms. PMID- 9855187 TI - Growth behavior of fibroblasts on microgrooved polystyrene. AB - We investigated the contact guidance phenomenon of rat dermal fibroblasts (RDF) on microgrooved polystyrene substrates. Grooves were 1 microm deep, and between 1 and 10 microm wide. Light microscopy and digital image analysis (DIA) showed that RDF were oriented on all microgrooved substrates. Scanning electron microscopy showed that RDF cultured on 1 or 2 microm wide grooves were positioned on top of the ridges. On the wider 5 and 10 microm grooves, the cells were able to descend into the grooves. In confocal laser scanning microscopy, focal adhesions were lying in the same direction as the actin filament where they attached to. DIA confirmed an orientational behavior of focal adhesions and actin filaments on microgrooves. There were no differences in the measured orientation between the different grooves. Besides, no obvious preference was found for focal adhesions to lie along edges of the surface ridges. Transmission electron microscopy showed that focal adhesions were able to bend along the edges of ridges. On the basis of our observations, we suggest that the breakdown and formation of fibrous cellular components, especially in the filopodium, is influenced by the microgrooves. The microgrooves create a pattern of mechanical stress, which influences cell spreading and cause the cell to be aligned with surface microgrooves. PMID- 9855188 TI - A novel surgical glue composed of gelatin and N-hydroxysuccinimide activated poly(L-glutamic acid): Part 1. Synthesis of activated poly(L-glutamic acid) and its gelation with gelatin. AB - Although fibrin glue has been widely used as a surgical adhesive, its components, fibrinogen and thrombin, obtained from human blood are not completely free from the risk of virus infection due to acquired immune deficiency and hepatitis. Recently, we have reported that a polymer pair composed of gelatin and poly(L glutamic acid) (PLGA) promptly forms a gel and can firmly bond to soft tissues when crosslinked with the aid of water-soluble carbodiimide (WSC). The present study was undertaken to design a new PLGA-gelatin glue without using WSC. Two kinds of PLGA with molecular weights of 71 and 22 kDa were employed to prepare N hydroxysuccinimide (NHS) activated derivatives. The NHS-activated PLGA could be synthesized at high yields and was found to be stable for an extended time without losing the ability to crosslink with gelatin when stored under a dry-cold condition. This NHS-activated PLGA could spontaneously form a gel with gelatin in an aqueous solution within a short time, comparable to a commercial fibrin glue, when gelation was allowed to proceed at pH 8.3. The NHS-activated PLGA prepared from PLGA with the molecular weight of 22 kDa could be readily dissolved at high concentrations and its ability to form a gel was maintained for more than 10 min when an acidic 8% NHS-activated PLGA solution was used. The bonding strength of PLGA gelatin glues with natural tissue was higher than that of fibrin glue. These findings strongly suggest that this combination of gelatin and NHS-PLGA is very promising as a surgical adhesive and may possibly replace fibrin glues prepared from human blood components. PMID- 9855189 TI - A liposomal hydrogel for the prevention of bacterial adhesion to catheters. AB - The adhesion of bacteria to medical implants and the subsequent development of a biofilm frequently results in the infection of surrounding tissue and may require removal of the device. We have developed a liposomal hydrogel system that significantly reduces bacterial adhesion to silicone catheter material. The system consists of a poly (ethylene glycol)-gelatin hydrogel in which liposomes containing the antibiotic ciprofloxacin are sequestered. A poly (ethylene glycol) gelatin-liposome mixture was applied to a silicone surface that had been pre treated with phenylazido-modified gelatin. Hydrogel cross-linking and attachment to surface-immobilized gelatin was accomplished through the formation of urethane bonds between gelatin and nitrophenyl carbonate-activated poly (ethylene glycol). Liposomal hydrogel-coated catheters were shown to have an initial ciprofloxacin content of 185+/-16 microg cm(-2). Ciprofloxacin was released over seven days with an average release rate of 1.9+/-0.2 microg cm(-2) h(-1) for the first 94 h. In vitro assays using a clinical isolate of Pseudomonas aeruginosa established the antimicrobial efficacy of the liposomal hydrogel. A modified Kirby-Bauer assay produced growth-inhibition zone diameters of 39+/-1 mm, while bacterial adhesion was completely inhibited on catheter surfaces throughout a seven-day in vitro adhesion assay. This new antimicrobial coating shows promise as a prophylactic and/or treatment for catheter-related infection. PMID- 9855190 TI - Molecular barriers to biomaterial thrombosis by modification of surface proteins with polyethylene glycol. AB - For cardiovascular biomaterials, thrombosis, thromboembolism and vascular graft occlusion are believed to be precipitated by the adsorption of proteins containing adhesive ligands for platelets. Polyethylene-glycol-diisocyanate (PEG diisocyanate, 3400 MW) may potentially react with protein amines to form molecular barriers on adsorbed proteins on biomaterials, thereby masking adhesive ligands and preventing acute surface thrombosis. To test this notion, PE, PTFE, and glass microconduits were pre-adsorbed with fibrinogen and treated with PEG diisocyanate, non-reactive PEG-dihydroxyl, or remained untreated. Following perfusion of 111In-labeled platelets in whole human blood for 1 min (wall shear rate = 312 s(-1)), PEG-diisocyanate treated surfaces experienced 96% (PE), 97% (PTFE) and 94% (glass) less platelet deposition than untreated surfaces. Similar reductions were seen for PEG-diisocyanate versus PEG-dihydroxyl treatment. Low shear perfusions of plasma for 1 h prior to blood contact did not reduce the inhibitory effect of PEG-diisocyanate. Platelet adhesion onto collagen-coated glass coverslips and platelet deposition onto preclotted Dacron were also reduced by treatment with PEG-diisocyanate (93 and 91%, respectively). Protein-reactive PEG may thus have utility in forming molecular barriers on surface-associated proteins to inhibit acute thrombosis on cardiovascular biomaterials. PMID- 9855191 TI - Osteogenic response of rabbit tibia to hydroxyapatite particle-Plaster of Paris mixture. AB - A mixture of hydroxyapatite (HA) particles and plaster of Paris (POP) with saline solution was injected into the bone marrow of 30 unilateral tibiae of 30 rabbits. As a control, POP with saline solution was similarly injected into the bone marrow of 30 unilateral tibiae of 30 additional rabbits. Five rabbits in each group were killed 2, 4, 8, 12, 24 and 40 weeks after injection. Histological examination was performed on three tibiae of five killed animals, and X-ray diffraction analysis of the bone marrow was performed on the remaining two tibiae. In two of three tibiae, bridges of new bone were formed between the hydroxyapatite particles and the cortical bone, 4 weeks after the injection of the HA particle-POP mixture. New bone formation was seen in all three tibiae at 8 weeks and thereafter. No new bone formation was observed in the tibiae of the control group. The affinity index, a parameter of osteogenesis, was 0% at 2 weeks, 52.2+/-14.0% at 4 weeks, 74.8+/-5.3% at 8 weeks, 86.1+/-6.1% at 12 weeks, 90.1+/-8.3% at 24 weeks and 92.8+/-4.1% at 40 weeks after injection. Statistically significant increases were found between the affinity indexes of 2 weeks and 4 weeks and between those of 4 weeks and 8 weeks, p < 0.03 and p < 0.05, respectively. Between 12, 24 and 40 weeks, the indexes showed tendencies toward increases. X-ray diffraction of bone marrow of two tibiae into which HA particle-POP mixture or POP alone was injected showed that POP had disappeared 8 weeks after injection. PMID- 9855192 TI - Osteogenic response of the rabbit femur to a hydroxyapatite thermal decomposition product-fibrin glue mixture. AB - The effect of fibrin glue on the hardening process of hydroxyapatite thermal decomposition product (HATDP) was investigated in vitro. To study, in vivo, the effect of fibrin glue on osteogenesis, a mixture of HATDP with fibrin glue was injected into the left femurs of 20 rabbits, and HATDP without fibrin glue was injected into the right femurs. New bone formation was observed in the area of direct contact with the fibrin-containing mixture in the metaphysis at 2 weeks after injection in two of four femurs, but not in the fibrin-free group. At 4 weeks after injection, new bone formed a bridge between the mixture and the cortical bone in three of the four specimens. A parameter of osteogenesis (the affinity index) was calculated at 4, 12 and 24 weeks after injection. It was respectively 0.14+/-0.02, 0.49+/-0.12 and 0.44+/-0.05 for the fibrin-containing mixture, and 0.14+/-0.062, 0.44+/-0.15 and 0.49+/-0.04 for the control. A comparable but significant increase in the average affinity index was noted over 12 weeks in both the fibrin-containing and the fibrin-free HATDP. These findings indicate that fibrin glue accelerated the hardening of HATDP but did not inhibit the process of osteogenesis. PMID- 9855193 TI - Prosthesis-bone interface. AB - In this overview, which is based on selected books and reviews, the microscopic appearances of prosthesis-bone interfaces resulting from clinically relevant implantation techniques are highlighted. The following techniques are distinguished to insert and attach prostheses in the recipient bone: impaction into bone, primary mechanical interlocking, cement fixation, bone ongrowth and secondary mechanical interlocking with bioinert materials, and enhanced bone ongrowth and bone bonding to bioactive materials. The resulting typical histomorphologies of orthopedic and dental prostheses-bone interfaces are described and illustrated from the author's studies employing light, fluorescence, and backscattered electron microscopy, and corresponding microradiographs of undecalcified ground sections of bone and implants. Special emphasis is given to the mineralization-demineralization kinetics of the interfacial bone matrix interacting with specific surface reactions of some implant materials. Consequently, the distinction between bioinert and bioactive prosthetic materials is critically analyzed. PMID- 9855194 TI - Recombinant human bone morphogenetic protein-2 and collagen for bone regeneration. AB - The study reported describes a combination of recombinant human bone morphogenetic protein-2 (rhBMP-2) and collagen (C) to regenerate bone. Unilateral critical-sized defects (CSDs) were prepared in radii of 32 skeletally mature New Zealand white rabbits. Rabbits were divided evenly among four treatments: autograft, absorbable C (Helistat), 35 microg of rhBMP-2 combined with absorbable C (rhBMP-2/C), and untreated CSDs. The two euthanasia periods were 4 and 8 weeks. Radiographs were taken the day of surgery, every 2 weeks, and at term and the percent of radiopacity was measured. Data analysis revealed a time-dependent increase in the percent radiopacity with rhBMP-2/C. Histological examination revealed the rhBMP-2/C treatment regenerated osseous contour by 8 weeks. According to quantitative histomorphometry, the CSD and C groups had significantly less new bone than either autograft or rhBMP-2/C (p < or = 0.05). The results suggest that rhBMP-2/C could be an effective therapy to restore segmental bone defects. PMID- 9855195 TI - Radiomorphometry and biomechanical assessment of recombinant human bone morphogenetic protein 2 and polymer in rabbit radius ostectomy model. AB - The study objective was to determine the mechanical integrity and radiopacity of regenerated bone within critical-sized defects (CSDs) in radii of rabbits using recombinant human bone morphogenetic protein 2 (rhBMP-2) with a porous, biodegradable poly(D,L-lactic acid) (PDLLA) carrier (designated PLA). Twenty millimeter, unilateral radial ostectomies were created in 96 skeletally mature New Zealand white rabbits. The rabbits were randomly assigned to six treatment groups with two euthanasia periods. Treatment groups included unfilled defect (n = 8), segmental autograft (n = 8), PLA + 0 microg rhBMP-2 (n = 8), PLA + 17 microg rhBMP-2 (n = 8), PLA + 35 microg rhBMP-2 (n = 8), and PLA + 70 microg rhBMP-2 (n = 8). The radiopacity was significantly greater for the 35- and 70 microg rhBMP-2 groups at 4 weeks compared to unfilled controls, PLA only, and 17 microg rhBMP-2 groups and equivalent to the autograft. At 8 weeks all groups receiving rhBMP-2 were equivalent to the autograft and significantly greater than unfilled defects and PLA alone. Similarly, the biomechanical analysis indicated significantly greater torque at failure for the 35-microg rhBMP-2 group compared to all other groups at 4 weeks. By 8 weeks all groups receiving rhBMP-2 and autograft had significantly greater torque than unfilled controls and PLA alone. These radiomorphometric and biomechanical results indicate PLA may be a suitable carrier for rhBMP-2 used for skeletal regeneration. PMID- 9855196 TI - Preliminary in vivo report on the osteocompatibility of poly(anhydride-co-imides) evaluated in a tibial model. AB - A novel class of polymers with mechanical properties similar to cancellous bone are being investigated for their ability to be used in weight-bearing areas for orthopedic applications. The poly(anhydride-co-imide) polymers based on poly[trimellitylimidoglycine-co-1,6-bis(carboxyphenoxy)hexan e] (TMA-Gly:CPH) and poly[pyromellitylimidoalanine-co-1,6-bis(carboxyphenoxy)hexa ne] (PMA-Ala:CPH) in molar ratios of 30:70 were investigated for osteocompatibility, with effects on the healing of unicortical 3-mm defects in rat tibias examined over a 30-day period. Defects were made with surgical drill bits (3-mm diameter) and sites were filled with poly(anhydride-co-imide) matrices and compared to the control poly(lactic acid-glycolic acid) (PLAGA) (50:50), a well-characterized matrix frequently used in bone regeneration studies, and defects without polymeric implants. At predetermined time intervals (3, 6, 9, 12, 20, and 30 days), animals were sacrificed and tissue histology was examined for bone formation, polymer tissue interaction, and local tissue response by light microscopy. The studies revealed that matrices of TMA-Gly:CPH and PMA-Ala:CPH produced responses similar to the control PLAGA with tissue compatibility characterized by a mild response involving neutrophils, macrophages, and giant cells throughout the experiment for all matrices studied. Matrices of PLAGA were nearly completely degraded by 21 days in contrast to matrices of TMA-Gly:CPH and PMA-Ala:CPH that displayed slow erosion characteristics and maintenance of shape. Defects in control rats without polymer healed by day 12, defects containing PLAGA healed after 20 days, and defects containing poly(anhydride-co-imide) matrices produced endosteal bone growth as early as day 3 and formed bridges of cortical bone around matrices by 30 days. In addition, there was marrow reconstitution at the defect site for all matrices studied along with matured bone-forming cells. This study suggests that novel poly(anhydride-co-imides) are promising polymers that may be suitable for use as implants in bone surgery, especially in weight-bearing areas. PMID- 9855197 TI - Principles of tissue engineering applied to programmable osteogenesis. AB - This article presents a strategy for design, engineering, and fabrication of a bioresorbable, manufactured bone graft substitute (BGS) device. The approach is based on established precepts of osteogenesis, molecular biology of hyaluronic acid and osteoinductive proteins, and theoretical preformance criteria for such a device collated from the literature of 1991 to 1996. Application of this design and engineering strategy results in a composite device consisting of a D,D-L,L polylactic acid macrostructure optimized to the architecture of cancellous bone, a microstructure composed of a filamentous velour of hyaluronan and a recombinant human bone morphogenetic protein 2 (rhBMP-2). The performance of this construct was tested in vivo in the dog, intertransverse process, spinal fusion model and in a critical sized defect of the rabbit radius. Data from these studies are used to illustrate principle points of the design and engineering concept. PMID- 9855198 TI - Resorbable calcium phosphate bone substitute. AB - The in vitro and in vivo properties of a novel, fully resorbable, apatitic calcium phosphate bone substitute (ABS) are described. The ABS was prepared from calcium phosphate precursors that were hydrated to form an injectable paste that hardens endothermically at 37 degrees C to form a poorly crystalline apatitic calcium phosphate (PCA). The PCA reaction product is stable in vivo as determined by FTIR and XRD analysis of rabbit intramuscular implants of ABS retrieved 4, 7, and 14 days postimplantation. Bone formation and resorption characteristics of the ABS material were characterized in a canine femoral slot defect model. Femoral slot defects in dogs were filled with either autologous bone implants or the ABS material. Sections of femoral bone defect site from animals sacrificed at 3, 4, 12, 26, and 52 weeks demonstrated that new bone formation proceeded similarly in both autograft and ABS filled slots. Defects receiving either material were filled with trabecular bone in the first 3 to 4 weeks after implantation; lamellar or cortical bone formation was well established by week 12. New bone formation in ABS filled defects followed a time course comparable to autologous bone graft filled defects. Histomorphometric evaluation of ABS resorption and new bone formation indicated that the ABS material was greater than 99% resorbed within 26 weeks; residual ABS occupied 0.36+/-0.36% (SEM, n = 4) of the original defect area at 26 weeks. Quantitatively and qualitatively, the autograft and ABS were associated with similar new bone growth and defect filling characteristics. PMID- 9855199 TI - Approaches to study of ischemia in bone. AB - Ischemic osteonecrosis is a complication of certain traumatic and a number of idiopathic conditions. The course of the disease may result in collapse of the convex member of a joint and osteoarthritis, often requiring arthroplasty. Increasing incidence in young adults poses a challenge for development of long term joint prostheses. Current status of research into the disease is discussed and three new models using intravital microscopy described. The first, an arterial occlusion (AO) model, creates ischemia by occluding the common iliac artery exclusively, avoiding direct trauma on other tissues in the limb. The second, a total occlusion (TO) model utilizes classical tourniquet occlusion of the thigh vessels. The third, a venous occlusion (VO) model, is also a tourniquet procedure but it blocks occlusion of the femoral artery with a protective sheath. Preliminary results from AO and TO studies are reported which show that reperfusion injury is detectible after ischemia doses as short as 4 h. This complication was confirmed by observation of leukocyte adherence, secondary ischemia, and abnormal vessel leakage. Also, a new quantitation of osteonecrosis is introduced whereby fluorescently-tagged dead osteocytes and computer-based image processing provide values for an "osteonecrosis index." Viewing of all vascular events is made possible by intravital microscopy through a bone chamber window implanted in rabbit tibias. It is proposed that such chronic visual techniques allow quantitation of events leading to osteonecrosis as well as the revascularization, resorption and bone apposition of creeping substitution which characterizes postischemia recovery. PMID- 9855200 TI - Polymer concepts in tissue engineering. AB - Traumatic injuries, cancer treatment, and congenital abnormalities are often associated with abnormal bone shape or segmental bone loss. Restoration of normal structure and function in these cases requires replacement of the missing bone that may be accomplished by surgical transfer of natural tissue from an uninjured location elsewhere in the body. However, this procedure is limited by availability, adequate blood supply, and secondary deformities at the donor site. One strategy to overcome these problems is to develop living tissue substitutes based on synthetic biodegradable polymers. Three methods of bone regeneration using biodegradable polymers are being studied in our laboratory: tissue induction, cell transplantation, and fabrication of vascularized bone flaps. Injectable polymers are used for filling skeletal defects and guiding bone tissue growth. Their main advantage is minimizing the surgical intervention or the severity of the surgery. Polymer-cell constructs also hold great promise in the field of tissue engineering. They provide a scaffold on which cells grow and organize themselves. As the cells begin to secrete their own extracellular matrix, the polymer degrades and is eventually eliminated from the body, resulting in completely natural tissue replacement. Bone flaps can be fabricated ectopically into precise shapes and sizes. With an attached vascular supply, these flaps can be transferred into areas deficient in vascularity. This article discusses polymer concepts regarding bone tissue engineering and reviews recent advances of our laboratory on guided bone regeneration using biodegradable polymer scaffolds. PMID- 9855201 TI - BoneSource hydroxyapatite cement: a novel biomaterial for craniofacial skeletal tissue engineering and reconstruction. AB - BoneSource-hydroxyapatite cement is a new self-setting calcium phosphate cement biomaterial. Its unique and innovative physical chemistry coupled with enhanced biocompatibility make it useful for craniofacial skeletal reconstruction. The general properties and clinical use guidelines are reviewed. The biomaterial and surgical applications offer insight into improved outcomes and potential new uses for hydroxyapatite cement systems. PMID- 9855202 TI - Effect of modification of oxide layer on NiTi stent corrosion resistance. AB - Because of its good radiopacity, superelasticity, and shape memory properties, nickel-titanium (NiTi) is a potential material for fabrication of stents because these properties can facilitate their implantation and precise positioning. However, in vitro studies of NiTi alloys report the dependence of alloy biocompatibility and corrosion behavior on surface conditions. Surface oxidation seems to be very promising for improving the corrosion resistance and biocompatibility of NiTi. In this work, we studied the effect on corrosion resistance and surface characteristics of electropolishing, heat treatment, and nitric acid passivation of NiTi stents. Characterization techniques such as potentiodynamic polarization tests, scanning electron microscopy, Auger electron spectroscopy, and X-ray photoelectron spectroscopy were used to relate corrosion behavior to surface characteristics and surface treatments. Results show that all of these surface treatments improve the corrosion resistance of the alloy. This improvement is attributed to the plastically deformed native oxide layer removal and replacement by a newly grown, more uniform one. The uniformity of the oxide layer, rather than its thickness and composition, seems to be the predominant factor to explain the corrosion resistance improvement. PMID- 9855203 TI - Microstructural and in vitro chemical investigations into plasma-sprayed bioceramic coatings. AB - Hydroxyapatite (HA) coatings plasma sprayed without and with bond coats (titania, zirconia) onto titanium alloy (Ti6A14V) substrates under both atmospheric and low pressure plasma spray conditions were investigated in terms of their microstructure and their resorption resistance during immersion in simulated body fluid (Hank's balanced salt solution). The microstructures of test samples were characterized using SEM on as-sprayed and leached surfaces and on the corresponding cross sections. Selected coating systems were studied by 2 dimensional secondary ion mass spectroscopy imaging to obtain information on plasma spray induced diffusional processes at the coating interfaces, as well as the spatial distribution of minor and trace elements. Coatings consisting of thin (10-15 microm) titania/zirconia (eutectic ratio) and titania bond coats, combined with a 150- to 180-microm thick HA top coat, yielded peel strengths in excess of 32 N/m, as well as sufficient resorption resistance. PMID- 9855204 TI - Histological, chemical, and crystallographic analysis of four calcium phosphate cements in different rabbit osseous sites. AB - Four calcium phosphate cement formulations were implanted in the rabbit distal femoral metaphysis and middiaphysis. Chemical, crystallographic, and histological analyses were made at 2, 4, and 8 weeks after implantation. When implanted into the metaphysis, part of the brushite cement was converted into carbonated apatite by 2 weeks. Some of the brushite cement was removed by mononuclear macrophages prior to its conversion into apatite. Osteoclastlike cell mediated remodeling was predominant at 8 weeks after brushite had converted to apatite. The same histological results were seen for brushite plus calcite aggregate cement, except with calcite aggregates still present at 8 weeks. However, when implanted in the diaphysis, brushite and brushite plus calcite aggregate did not convert to another calcium phosphate phase by 4 weeks. Carbonated apatite cement implanted in the metaphysis did not transform to another calcium phosphate phase. There was no evidence of adverse foreign body reaction. Osteoclastlike cell mediated remodeling was predominant at 8 weeks. The apatite plus calcite aggregate cement implanted in the metaphysis that was not remodeled remained as poorly crystalline apatite. Calcite aggregates were still present at 8 weeks. There was no evidence of foreign body reaction. Osteoclastlike cell remodeling was predominant at 8 weeks. Response to brushite cements prior to conversion to apatite was macrophage dominated, and response to apatite cements was osteoclast dominated. Mineralogy, chemical composition, and osseous implantation site of these calcium phosphates significantly affected their in vivo host response. PMID- 9855205 TI - Interface strength studies of calcium phosphate ceramic coated strain gauges. AB - In vivo strain gauging has been used to understand physiological loading and bone remodeling. In early studies, a cyanoacrylate adhesive was used to bond gauges to bone, even though this adhesive is susceptible to biodegradation that results in rapid debonding. Calcium phosphate ceramic (CPC) coated gauges have been successfully bonded to bone for long periods. However, earlier studies noted occasional debonding of coatings from gauges. The goals of this project were to develop a technique to securely bond particles to gauge backings and develop an in vitro test and assess its accuracy in simulating in vivo degradation of this interface. Gauges were heated for different time intervals, roughened with carbide papers, and prepared using layered coatings of polysulfone and CPC particles that varied in size, shape, and crystallinity. They were soaked in solution or placed in muscle pouches of rats for up to 16 weeks. They were then epoxied to fixtures, mounted on an MTS machine, and loaded to failure. Heating and roughening gauge surfaces increased the interface strengths by up to 2000%. In vivo and in vitro testing showed an initial drop in the interface strength, which leveled off to approximately 7.0+/-2.0 MPa. PMID- 9855206 TI - Endothelial cell responses to chitin and its derivatives. AB - The effects of chitin and its derivatives on the proliferation of human umbilical vein endothelial cells (HUVECs) and on the production of cytokines were examined in vitro. Chitin and its derivatives had no effect on the proliferation of cultured HUVECs. N-Sulfonated 70% deacetylated chitin (S-DAC70) stimulated the production of interleukin (IL)-1beta, IL-6, IL-8, and tumor necrosis factor (TNF) alpha from HUVECs. Compared to S-DAC70, the other materials tested in the present study showed less effect in the stimulation of IL-8 and TNF-alpha production and had no effect in the stimulation of IL-1beta and IL-6 production. These results indicated that S-DAC70 affects HUVECs function but not proliferation. PMID- 9855207 TI - Pathologic supracondylar fracture due to osteolytic pseudotumor of knee following cementless total knee replacement. AB - Wear debris of polyethylene, polymethylmethacrylate, and metal have been recognized to be associated with foreign body reactions, osteolysis, and aseptic prosthetic loosening after joint replacement arthroplasty. Further, foreign body reaction due to the presence of extensive wear debris can cause aggressive granulomatous lesions and pathologic fracture. To our knowledge, there has been no previous report of pathologic fracture of the femur due to an agressive pseudotumor. This report describes a case of pathologic supracondylar fracture of the femur 6 years and 5 months after cementless total knee replacement arthroplasty. The fracture occurred through an aggressive expanding soft tissue mass that was a tumorlike lesion secondary to polyethylene wear debris. The lesion was associated with massive osteolysis around the femoral component of the total knee prosthesis. PMID- 9855208 TI - Sporadic inclusion-body myositis and its similarities to Alzheimer disease brain. Recent approaches to diagnosis and pathogenesis, and relation to aging. AB - Sporadic inclusion-body myositis (s-IBM) is the most common, debilitating and progressive muscle disease beginning at the age 50 or later. The most characteristic pathologic feature is vacuolar degeneration of muscle fibers accompanied by intrafiber congophilia and clusters ("tangles") of paired-helical filaments, containing phosphorylated tau. An unusual feature of sporadic inclusion-body myositis is accumulation within its abnormal muscle fibers of several proteins that are characteristic of Alzheimer disease brain, including epitopes of beta-amyloid precursor protein (betaAPP), phosphorylated tau, alpha-1 antichymotrypsin, apolipoprotein E, and presenilin-1. Indicators of oxidative stress are also present within abnormal s-IBM muscle fibers. In this review, we describe new advances seeking the pathogenic mechanism of sporadic inclusion-body myositis. We hypothesize on the possible pathogenic role of abnormally accumulated proteins, and we propose that important contributory factors leading to inclusion-body myositis are the milieu of muscle-fiber aging and oxidative stress. In addition, we present evidence that overexpression of adenovirus transferred betaAPP gene in cultured human muscle fibers induces aspects of the inclusion-body myositis phenotype, and suggest that betaAPP-overexpression is an early event in the pathogenic cascade causing inclusion-body myositis. PMID- 9855209 TI - Hip involvement in seropositive rheumatoid arthritis. Survivorship analysis with a 15-year follow-up. AB - In a prospective survey of recent arthritis 96 patients had erosive and seropositive rheumatoid arthritis (RA). Severe radiological changes in the hipjoint (Larsen grade 3-5) were observed in 31 (32%) and acetabular protrusion in five (5%) patients 15 years after the onset of arthritis. At the end of follow up Larsen score 0-100 of peripheral joints (p < 0.015), HAQ (p < 0.001), ESR (p < 0.001), and CRP (p < 0.01) were significantly higher in patients with hip joint destruction than in those without. Secondary amyloidosis was found in eight patients with and in three patients without hip involvement (p = 0.002). In survivorship analysis (the Larsen 0-2 grade hips) an overall predicted rate of survival was 80% after 14 years of rheumatoid arthritis. When analyzed according to HLA-B27 the rate of radiological survival for the patients with a positive test was 62% and 87% for the others (p = 0.02). In conclusion, RA patients with high number of destructive peripheral joints (Larsen score) and high ESR or CRP should be carefully monitored due to the increased risk of hip destruction. PMID- 9855210 TI - Emotional status in systemic lupus erythematosus. AB - Psychological and emotional status was assessed in 36 patients with systemic lupus erythematosus (SLE) with the General Health Questionnaire-30 (GHQ-30) and Minnesota Multiphasic Personality Inventory (MMPI) questionnaires. The two tests were found to measure different aspects of psychological functioning. More than 50% of the patients could be classified as cases with mild psychiatric disturbances, according to the GHQ, and 28% had an abnormal score > 70 on the MMPI depression subscale, indicating significant depression. Emotional disturbances such as problems with social functioning, personal discomfort in social situations, and depressive mood, were frequent and associated with skin and joint abnormalities. This suggests additional etiologies for psychological dysfunction among SLE patients other than the direct central nervous system (CNS) effect of SLE. PMID- 9855211 TI - Patients with primary osteoarthritis show no change with ageing in the number of osteogenic precursors. AB - The variation in marrow colony forming unit-fibroblastic (CFU-F) number in 59 patients (14-87 years of age) undergoing corrective surgery (14 controls; 14-48 years of age) or hip arthroplasty for primary osteoarthritis (45 OA; 46-87 years of age) was examined to determine whether marrow CFU-F, derived from marrow stromal fibroblastic stem cells, are maintained with the development of primary osteoarthritis (OA). Total colony number, colony size as well as alkaline phosphatase-positive colonies were determined. The mean fibroblast colony forming efficiency from the whole patient group was 2.4 x 10(-5) +/- 1.4 x 10(-5). Ageing had no effect on the colony forming efficiency or on the alkaline-phosphatase positive colony forming efficiency, irrespective of gender. Thus precursor cells with the potential for osteogenic differentiation are maintained in OA with ageing. However, colony size showed a significant reduction with age, implying altered proliferation potential of osteogenic progenitors with ageing. This ageing effect may not be as significant in OA as in the rest of the population as bone mineral density is often preserved in osteoarthritis. As there is no apparent deficit in primitive progenitor cells, this preservation may be the result of altered regulation of osteoprogenitor activity in OA. PMID- 9855212 TI - Treatment of "frozen shoulder" with distension and glucorticoid compared with glucorticoid alone. A randomised controlled trial. AB - This study is a comparison of treatments of idiopathic "Frozen Shoulder" (adhesive capsulitis), distension combined with steroid is compared with steroid alone. Evaluation was based on pain scales, analgesic usage, and range of motion outcome scales. Out of one-hundred twenty patients (age, mean 51, range 21-70) that were referred under the diagnosis FS, twenty-six fulfilled the criteria for inclusion in the study, but four patients did not want to participate in the trial, giving a total of 22 patients (age, mean 53, range 40-65) in the study. Patients were randomised by the envelope method. Two patients dropped-out, one in each treatment group thus leaving the study with 20 patients for the final statistical analysis. Eight were treated with steroid alone and 12 with distension combined with steroid. Patients received one treatment per week for a six weeks period with a follow-up at 12 weeks. They were evaluated by pain VAS on function and at rest within the study period, the different ranges of motion (ROM) were measured at inclusion time and subsequent afterwards at 3, 6, and 12 weeks. The VAS outcomes showed no difference between the treatments (VAS-function p=0,1; VAS-rest p=0.1), while in the distension group ROM showed significant improvement in all directions except extension (external p=0.0007, flexion p=0.03, extension p=0,01). The analgesic usage was significantly lower in the group treated with distension at the end of the study (p=0.008). A blinded clinical assessment of ROM also showed significant improvement (p=0.002). It is concluded that distension with steroid can seem to help in management of "Frozen Shoulder". Other studies seems to support the conclusion. PMID- 9855213 TI - Persistent or severe course of reactive arthritis following Salmonella enteritidis infection. A prospective study of 9 cases. AB - During a 7 year-period 9 children (7 boys, 2 girls) with juvenile reactive arthritis (JReA) due to Salmonella enteritidis (Se) were prospectively studied because of an unusual type of onset and/or course of the disease. The mean duration of JReA activity was 9 +/- 3.6 months. The mean follow-up time was 55.2 +/- 17.4 months. JReA presented as any of the three types of juvenile chronic arthritis (JCA), namely, as asymmetrical oligoarthritis, polyarthritis, or systemic JCA in 5, 2, and 2 patients respectively. Two patients had pericarditis and three developed the complete or incomplete Reiter's syndrome during the disease or during a recurrence. Five patients carried the HLA-B27 and 3/5 developed psoriatic lesions 1 to 15 months after the onset of JReA. The presence of HLA-B27 and psoriasis was associated with a more prolonged course of JReA. However, no patient developed late radiological lesions or sacroiliitis during follow-up. PMID- 9855214 TI - Low copy numbers of human T-cell lymphotropic virus type I (HTLV-I) tax-like DNA detected in the salivary gland of seronegative patients with Sjogren's syndrome in an HTLV-I endemic area. AB - To evaluate the hypothesis, proposed in previous reports from HTLV-I non-endemic areas, that HTLV-I is involved in a significant proportion, about a quarter, of Sjogren's syndrome patients who lack serum antibodies to the virus, we examined for the presence or absence of HTLV-I in DNA samples isolated from salivary gland tissues of 17 seronegative as well as 7 seropositive patients with Sjogren's syndrome in Nagasaki, Japan, where the virus is highly endemic. The nested two step polymerase chain reaction (PCR), with a sensitivity capable of detecting a single DNA molecule, failed to amplify the HTLV-I tax sequence from DNA of 14 of the 17 seronegative patients. The tax was only amplifiable from the tissue DNA of the remaining three seronegative patients. The detection rate, 3/17 (18%), was, unexpectedly, less than those previously reported from the HTLV-I non-endemic areas. Moreover, in contrast to high viral loads (10(-1) to 10(-3) per cell) in the salivary gland of the seropositive patients, a semiquantitative PCR revealed that the copy number of the HTLV-I tax in the gland tissue of these seronegative patients was very low, 10(-5) per cell. This level is unlikely to be sufficient to promote an inflammatory reaction in the tissue. Our findings might argue against the involvement of "prototype" HTLV-I in the pathogenesis of Sjogren's syndrome in seronegative patients. PMID- 9855215 TI - Carnitine and antioxidants levels in patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a heterogeneous disorder with a spectrum of clinical severity ranging from mild arthritis to a crippling joint disease with involvement of internal organs. Carnitine is essential for muscle energy production and is required for the transport of long chain fatty acids and the acyl coenzyme A derivatives across the inner mitochondrial membrane. The levels of malondialdehyde (MDA), an index of lipid peroxidation, and the antioxidants copper-zinc superoxide dismutase (CuZnSOD), glutathione (GSH), ceruloplasmin (CP), catalase (CAT), and carnitine were assessed in 42 patients with RA and 24 control subjects. While plasma carnitine and erythrocyte CuZnSOD levels were significantly lower in the patients with RA compared with the control group (p<0.01 and p<0.001, respectively), the CAT level was not different from controls (p>0.05). Plasma MDA, CP, and erythrocyte GSH levels were significantly higher than in the control group (p<0.001, p<0.001 and p<0.01, respectively). MDA levels showed a positive correlation with CP and GSH levels (r=0.716, p<0.001 and r=0.492, p<0.01, respectively). However, MDA, GSH, and CP demonstrated a negative correlation with carnitine (r=-0.719, p<0.001; r=-0.559, p<0.01, and r=-0.635, p<0.001, respectively) in the patient group but not in controls. There was also a significant positive correlation between CP and GSH levels (r=0.561, p<0.01). However, neither CuZnSOD nor CAT levels demonstrated correlation withcarnitine, MDA, GSH, or CP levels. It was interesting that CAT activity was not altered and CuZnSOD activity decreased when compared with the control group. These results suggest that while CP, MDA and GSH levels increased, carnitine and CuZnSOD levels decreased, but CAT activity was unchanged. PMID- 9855216 TI - In vitro synthesis and characterization of a cartilaginous meniscus grown from isolated temporomandibular chondroprogenitor cells. AB - Internal derangement of the temporomandibularjoint can lead to perforation of the intra-articular meniscus and osteoarthritic degeneration. Current methods of repairing damaged menisci are limited by lack of biological compatibility of graft materials. This project aimed to synthesise and characterise a primate cartilaginous meniscus in vitro from harvested mandibular chondroprogenitor cells. Isolated cells from the mandibular cartilage of 12 young adult marmosets, aged 9-12 months, were grown in monolayer culture. After 21 days confluent colonies were resuspended and dispersed into a unpolymerised solution of type I collagen and fibrinogen. The resultant cell suspension was infiltrated into a resorbable type I collagen sponge carrier and allowed to polymerise. Aliquots of the cell-infiltrated sponge were maintained in organ culture for a further 14 days. Cultures were characterised using histochemical and immunocytochemical localisation of collagen and proteoglycan species. Two-thirds of cells in confluent 21-day monolayers expressed cartilage-specific type II collagen and chondroitin-4-sulphate. After 35 days organ cultures had formed a viable, organised, three-dimensional tissue mass consisting of mature chondrocytic cells interspersed in a dense cartilaginous matrix. The cartilaginous tissue generated in vitro may have potential application in the repair or replacement of damaged menisci in vivo. PMID- 9855217 TI - Cortical photostimulation with filtered visible light as a treatment for rheumatoid arthritis. AB - Not all extraocular photoreceptors in the human brain have been identified or their functions determined. The efferent and afferent fibres between the orbital frontal cortex and the hypothalamus cannot be overlooked in the context of the neuro-endocrine-immune feedback loop. It is suggested here that there are extraocular photoreceptors in the orbital frontal cortex, conceivably crucial for immunoregulation. The orbital frontal cortex of 7 volunteers with rheumatoid arthritis (RA) was photostimulated through the roof of the orbits with filtered visible light in sessions of 12, 14 and 16 minutes for a period of 6 weeks. Within 5 weeks, a significant decrease in the rheumatoid factor titres of 6 subjects was observed together with marked clinical improvement in 4 cases. PMID- 9855218 TI - Adult onset Still's disease associated with Epstein-Barr virus infection in a 66 year-old woman. AB - Adult onset Still's disease (AOSD) is a distinct clinical entity which affects predominantly young adults aged 16-35 years. Onset in elderly individuals is exceptional. Several reports have suggested a viral trigger in the pathogenesis of this disease. We describe a 66-year-old woman who fulfilled the proposed diagnostic criteria of AOSD and suffered concurrently from acute Epstein-Barr virus (EBV) infection. PMID- 9855219 TI - Analysis of epitope spreading over an eleven-year period in a patient with systemic lupus erythematosus. AB - During a period of more than eleven years serum samples of a patient with Systemic Lupus Erythematosus were collected and analyzed for anti-nuclear autoantibodies. High titer of anti-La/SS-B were detectable in all serum samples. The La/SS-B epitopes remained constant. Besides anti-La/SS-B antibodies all serum samples contained traces of anti-Ro/SS-A including anti-Ro52 and anti-Ro60 antibodies. During disease flares anti-Ro/SS A antibodies were upregulated and anti-dsDNA antibodies appeared, thus supporting the concept of an antigen driven intermolecular epitope spreading to Ro/SS-A and dsDNA. PMID- 9855220 TI - Fatal interstitial pneumonia due to cytomegalovirus following cyclophosphamide treatment in a patient with systemic lupus erythematosus. PMID- 9855221 TI - Tribological tests cannot replace clinical trials. PMID- 9855222 TI - Low wear and friction in alumina/alumina total hip joints: a hip simulator study. AB - We studied the wear of state-of-the-art alumina/alumina total hip joints with the uniaxial five-station, and the three-axis single-station hip joint simulators of Helsinki University of Technology. The diameters of the joints were 26, 28 and 32 mm. Visual examination and weighing of the components showed no wear. Scanning electron microscopy showed that (a) the wear marks were slight, consisting of removal of grains, multidirectional grooving and mild abrasion, (b) the wear mechanisms were similar to those seen in components removed from patients, and (c) there was no marked difference in the specimens worn in the uniaxial vs. three-axis simulator. The coefficient of friction was measured throughout the 5 million cycle wear test with the three-axis simulator. The average value was 0.007. We conclude that alumina/alumina joints had low wear and friction under the present test conditions. The results are promising, since minimal wear is one of the prerequisites for long-term success in prosthetic joints. PMID- 9855223 TI - Metal release from total hip articulations in vitro: substantial from CoCr/CoCr, negligible from CoCr/PE and alumina/PE. AB - We used a hip joint simulator to compare the metal release from CoCr/CoCr, CoCr/PE, and alumina/PE total hip articulations. The metal release was quantified by analyzing the Co, Cr, and Ni contents of the bovine serum lubricant used with atomic absorption spectroscopy. CoCr/CoCr articulations released substantial amounts of metal, whereas CoCr/PE was equal to the control, alumina/PE, in that metal release was negligible. The metal release was in accordance with the known clinical wear rates of CoCr/CoCr articulations. The largest dimensional changes occurred in polyethylene cups, the penetrations of CoCr heads to the polyethylene cups being twice that of the alumina head, which is consistent with clinical experience. The research on the wear behavior of different materials, aiming to find a prosthesis with negligible wear, needs to be continued. Due to the substantial metal release, the CoCr/CoCr articulation is hardly the final solution of the wear problem in total hip arthroplasty. PMID- 9855224 TI - Different cup migration in rheumatoid arthritis and arthrosis: a radiographic analysis of 127 uncemented acetabular cups. AB - We compared retrospectively the radiographic migration profiles of 82 acetabular components in 61 patients having rheumatoid arthritis with those of 45 hips having arthrosis who underwent a standardized technique of cementless arthroplasty with the Zweymuller prosthesis (Alloclassic). We used a modification of Dickob's technique of digital migration analysis that corrects for magnification errors and horizontal pelvic tilt. The rheumatoid patients were stratified as having oligoarticular, polyarticular, or mutilating arthritis. The overall rate of acetabular loosening in rheumatoid hips after mean 88 (26-117) months was 4%. Loosening was seen only in cases with mutilating arthritis and acetabular protrusion, where the direction of cup migration was also clearly different from that detected in the other types of rheumatoid arthritis and in arthrosis. The different patterns of cup migration in cementless hip replacement for rheumatoid arthritis, depending on disease severity, is of importance when comparing outcome of total hip arthroplasty in rheumatoid patients. PMID- 9855225 TI - Biases in a randomized comparison of three types of screw fixation in displaced femoral neck fractures. AB - We studied fixation of displaced femoral neck fractures prospectively in a randomized multicenter study, comparing 2 Olmed screws, 2 Tronzo screws and 3 Ullevaal hip screws. The study population consisted of 482 women and 125 men, of whom 432 women and 100 men were older than 65 years of age. Their median age was 80 (54-97) years. Despite agreement on criteria, the rates of reoperations for pain and failure--salvage (prosthesis replacement) and other reoperations (removal of implant)--differed significantly between the 3 hospitals regardless of type of fixation. In total, the percentages of salvage operations were: Olmed screw 17/175, Tronzo 17/130 and Ullevaal screw 11/302 (n.s.); the percentages of other reoperations were 11, 6 and 13, respectively (n.s.). In the whole series, the 2-year rate of salvage operations was 14%. No differences between the implants were found in patients older than 65 years of age. We conclude that an agreed, common definition of a hard end-point (reoperation) does not ensure comparability of results, because of differences in clinical decision making. PMID- 9855226 TI - Revision of unicompartmental knee arthroplasty: outcome in 1,135 cases from the Swedish Knee Arthroplasty study. AB - From 1975 through 1995, 45,025 knee arthroplasties were recorded in the prospective Swedish Knee Arthroplasty study. By the end of 1995, 1,135 of 14,772 primary unicompartmental knee arthroplasties (UKA) for localized, mainly medial arthrosis had been revised. The Marmor/Richards and St. Georg sledge/Endo-Link prostheses were used in 65%. Mean age at revision was 72 (71) years. 232 revisions were performed as an exchange UKA (partial in 97) and 750 as a total knee arthroplasty (TKA). 153 were revised by other modes. In medial UKA, the indication for revision was component loosening in 45% and joint degeneration in 25% and in lateral UKA, the corresponding figures were 31% and 35%, respectively. In 94 cases, unicompartmental components were added to the initially untreated compartment, in 14 with partial exchange of a component. The CRRR was estimated using survival statistics. After only 5 years, the risk of having a second revision was more than three times higher for failed UKAs revised to a new UKA (cumulative rerevision rate (CRRR 26%) than for those revised to a TKA (CRRR 7%). This difference remained, even if those revised before 1985, when modern operating technique was introduced, were excluded (CRRR 31% and 5%, respectively). UKA is a safe primary procedure, when performed with well-designed components and modern surgical technique. It gives documented good patient satisfaction, range of motion, pain relief and relatively few serious complications. However, once failed, the knee should be revised to a TKA. This applies to most modes of failure. Not even joint degeneration of the unoperated compartment can be safely treated by adding contralateral components; CRRR after this procedure was 17%, while it was 7% when converted to a TKA. PMID- 9855227 TI - Postoperative drainage of knee arthroplasty is not necessary: a randomized study of 90 patients. AB - We studied the management of postoperative drainage after total knee replacement (TKR). 90 primary total knee joint arthroplasties were prospectively randomized into 3 groups: a) no drain, b) an autotransfusion system, c) a standard disposable closed suction drainage system. We monitored hemoglobin and hematocrit values, drainage volume and transfusions (homologous and autologous), range of knee motion, knee swelling and hospital stay. Parameters were recorded preoperatively, days 0-8 and 4 months postoperatively. No significant differences were seen between the groups in any of the parameters measured. The results show no benefit from using postoperative drainage systems in knee arthroplasties. Savings of SEK 400 (USD 55) per patient would have resulted if drains had not been used at all. PMID- 9855229 TI - Tensile properties of the knee-joint capsule at an elevated intraarticular pressure. AB - In order to evaluate the capsular reaction to high intraarticular pressure (IAP) in the knee during arthroscopy, we examined 13 patients admitted for elective surgery with joint infusion to 30, 70, 120 and 170 mmHg IAP (4, 9.3, 16 and 22.7 kPa). The infusion was repeated once, at all IAP levels except 30 mmHg, after 2 min and was recorded for another 2 min. A pressure-time curve was recorded as a combined effect of viscoelastic properties of the capsule and extraarticular fluid absorption. 3 more knee joints were infused directly to 170 mmHg and a pressure-time curve was recorded for 15 min, after which time the joint was reinfused to 170 mmHg in order to estimate the change in joint volume due to absorption or relaxation. In 10 knees, the curves were uniform. At infusion, the pressure curve was almost linear above 70 mmHg. There were no signs of plastic deformation of the joint capsule at pressures below 120 mmHg, while at 170 mmHg there were signs of capsular deformation. At each pressure level, the curve revealed a rapid fall in initial pressure that gradually decreased because of capsular relaxation or fluid absorption. Repeated infusion delayed the fall in pressure, due to increased capsular stiffness. In 3 knees infused directly to 170 mmHg IAP, the slope for the first 2 min of the pressure-time curve did not differ from that found at maximal IAP in knees examined with stepwise increasing pressures. Discontinuity of the capsule, even of puncture size, influenced the pressure/volume correlation considerably. We conclude that at IAP levels of 170 mmHg, there are signs of plastic deformation of the joint capsule. In order to avoid capsular damage, knee arthroscopy should be done at intraarticular pressure levels below 120 mmHg. PMID- 9855228 TI - Inferior performance of Boneloc bone cement in total knee arthroplasty: a prospective randomized study comparing Boneloc with Palacos using radiostereometry (RSA) in 19 patients. AB - We compared Boneloc bone cement with conventional cement (Palacos) in fixating the tibial component during 2-5 years in 19 patients with gonarthrosis undergoing total knee arthroplasty in a prospective randomized study. Boneloc displayed significantly larger migration, subsidence and lift-off than Palacos. The difference was identifiable already within 3 months postoperatively, but became significant at 12 months. In the Boneloc group, all components showed subsidence of the posterior part and lift-off of the anterior part of the tibial component, whereas in the Palacos group, the locations of subsidence and lift-off were evenly distributed about the edge of the implant. At 5 years, both Boneloc knees so far investigated were clinical failures with high migration rates. We conclude that, even in total knee arthroplasty, there is a substantial risk that Boneloc leads to inferior clinical results, but later than in hip replacements. PMID- 9855230 TI - Patellar alignment evaluated by MRI. AB - We analyzed the congruence of the articular cartilage surfaces and the corresponding subchondral bone in the patellar joint. 20 volunteers underwent MRI investigations of the right patellar joint in 20 degree and 45 degree flexion in the axial plane. The sulcus, congruence, and lateral patellofemoral angles, measured on MRI slices centered through the midtransverse patella, were recorded. In 20 degree and 45 degree knee flexion, the bony sulcus and lateral patellofemoral angles were significantly different from the respective cartilagineous angle. We conclude that 1) measurement of the bony sulcus and lateral patellofemoral angles does not allow conclusions about the articular cartilage surface and its thickness, 2) the bony congruence angle corresponds well to the articular cartilage surface as an indicator of patellar centralization. PMID- 9855231 TI - Intramedullary nailing and functional bracing of tibial shaft fractures: 167 cases followed for minimum 2 years. AB - We treated 167 diaphyseal tibial fractures without reaming and with intramedullary fixation, using an unlocked Kuntscher nail. The patients were followed for a minimum of 2 years. The mean time to union was 16 weeks. There were no serious complications, but 1 case of nonunion, 1 case of deep infection and 2 cases of clinically relevant malalignment. Migration of the nail was observed in 11 cases. We conclude that this treatment is simple and gives satisfactory results in diaphyseal fractures after low- or mid-energy trauma. PMID- 9855232 TI - Scoring in forefoot surgery: a statistical evaluation of single variables and rating systems. AB - We assessed 13 scores for forefoot surgery and their single parameters in 200 cases of hallux surgery. Outcome expressed in descriptive terms from excellent to poor, as well as calculated as a numerical value differed to a great extent. The rank correlation between different scores showed poor conformity, in some combinations even slight negative correlations. The rank correlation between single parameters and the overall outcome gave good values for some variables included in most scores, e.g., walking distance, general activity, problems on uneven surfaces, shoe wear, cosmetics and pain. Little, if any, correlation was found for joint instability, range of motion of the MTP- and IP-joints, the use of walking aids and medication. We find a need for a generally accepted score, with parameters of high clinical relevance. PMID- 9855233 TI - Zanoli's procedure for overlapping fifth toe: retrospective study of 18 cases followed for 4-17 years. AB - Digitus quintus superductus pedis (overlapping fifth toe) is a congenital deformity, often bilateral, in which the fifth toe is medially directed, rotated and hyperextended. There is no consensus regarding the best treatment. We describe our experience using Zanoli's procedure, which is a tenodesis procedure using the extensor tendon of the fifth toe. Between 1979 and 1993, we operated on 16 patients (7 bilateral, total 23 feet). 13 (10 women) patients (18 feet), with a mean age of 26 (19-40) years, were seen on follow-up. The mean follow-up time was 8 (4-17) years. Pain relief was achieved in all patients. From the surgeon's point of view, 3 toes overcorrected due to technical errors were considered unsatisfactory. All patients, however, were satisfied with the result and would advise other patients to undergo the same operation. PMID- 9855234 TI - How well can a previous fracture indicate a new fracture? A questionnaire study of 29,802 postmenopausal women. AB - In a population-based, retrospective study, we investigated the effect of a previous fracture on the risk of a later fracture. A questionnaire was mailed to 46,353 postmenopausal women aged 50-80 years and 29,802 (64%) responded. Questions were asked about fractures after age 25 and in what year they had occurred. The fractures were grouped according to whether they occurred within 5 and 10 years before answering the questionnaire. The occurrence of a fracture preceding these time intervals increased the odds ratio (OR) of having sustained a hip fracture to 1.6 (95% CI 1.1-2.3) for a previous ankle fracture and to 3.5 (95% CI 2.4-5.0) for a previous humerus fracture. Corresponding figures for having sustained a spine fracture were 1.5 (95% CI 1.1-2.1) for a previous ankle fracture and 4.5 (95% CI 3.4-5.9) for a previous spine fracture. It seems possible to select women for evaluation and intervention against osteoporosis by using information on previous fragility fractures. PMID- 9855235 TI - Epidemiology of acute vertebral osteomyelitis in Denmark: 137 cases in Denmark 1978-1982, compared to cases reported to the National Patient Register 1991-1993. AB - We studied the epidemiology of acute, non-tuberculous, hematogenous vertebral osteomyelitis in Denmark during 1978-1982. 137 patients fulfilled the criteria for acute vertebral osteomyelitis. The incidence was 5/mill/year. There were no cases in the age group 20-29 years. The highest incidence was between 60-69 years (18/mill/year). The prevalence was 15 cases. The mean duration of the disease was 7 months. The lumbar spine was affected in 59%, the thoracic spine in 33% and the cervical spine in 8% of the cases. Insulin-dependent diabetes and treatment with systemic corticosteroids seemed to be significant risk factors, but not rheumatoid arthritis and abuse of alcohol or intravenous drugs. We found no demographic variables of importance for the incidence. In 46%, a primary focus was identified, urinary tract infection being the commonest. According to the National Patient Register 1991-1993, the relative number of reported patients with vertebral osteomyelitis had increased in the age group 20-49 years, compared to 1978-1982, but the incidence was highest in the group aged 60-79 years. PMID- 9855236 TI - Osteomyelitis treated with gentamicin-PMMA beads: 100 patients followed for 1-12 years. AB - We treated 100 patients having osteomyelitis with debridement and gentamicin-PMMA beads and followed them for 5 (1-12) years. 66 of the infections were chronic, in 18 cases combined with arthritis and in 3 cases with pseudarthrosis. They underwent 117 "treatment periods", consisting of one or more operations (total 152), in most cases with an interval of 2 weeks. No systemic antibiotics were necessary besides the local antibiotic treatment in 52 of the treatment periods. Healing was achieved in 92 patients, in 78 after a single treatment period which included 1-5 operations, in 14 after two or three treatment periods. Healing was more difficult to achieve when the infection was chronic, especially with a duration of more than 6 years or when caused by elective surgery. Local antibiotic treatment with gentamicin PMMA beads has the advantage that the wound can be closed primarily and that a higher local antibiotic concentration in the tissues can be achieved, often making systemic antibiotic treatment unnecessary. PMID- 9855237 TI - Slime production of Staphylococcus epidermidis: increased bacterial adherence and accumulation onto pure titanium. AB - In an in vitro study using Staphylococcus epidermidis RP 62 A, a slime-producing strain and its isogenic slime-negative mutant M7, we demonstrated that both strains adhere to pure titanium discs with significantly higher colony counts for the slime-producing strain. The colony count was dependent on temperature, time and strain. Prolonged incubation time (24 h) under growth conditions leads to higher colony counts for the slime-producing strain RP 62 A. As the slime negative mutant M7 can adhere to, but not form multiple layers on metallic surfaces, increase of incubation time does not produce higher colony counts on the metallic surface. We conclude that slime production is important for adherence and subsequent accumulation of S. epidermidis onto pure titanium discs in vitro. PMID- 9855238 TI - Periosteum and bone marrow in bone lengthening: a DEXA quantitative evaluation in rabbits. AB - We quantitatively studied the role of periosteum and bone marrow-endosteum during lengthening in 18 growing rabbits, comparing four surgical procedures: 1) periosteum and bone marrow preservation, 2) periosteum preservation, bone marrow destruction, 3) periosteum destruction, bone marrow preservation, 4) periosteum and bone marrow destruction. An external fixator was set on one femur, the other serving as a control. Distraction began on day 5 and stopped on day 25 (0.25 mm/12 hours). On day 30, femora were harvested with a layer of muscle. Area, bone mineral content and density were measured by dual-energy x-ray absorptiometry. Procedure 2 showed the highest increase in bone mineral content around the elongated callus (127%) compared to procedures: 1 (81%), 3 (25%) and 4 (-8%, i.e., resorption of bone ends). A statistically significant effect on bone formation was observed when preserving (vs. destroying): 1) periosteum, 2) bone marrow (effect observed only around the distraction gap), 3) periosteum and bone marrow in combination. Periosteum alone forms a larger callus, with more mineral content than bone marrow alone, and destruction of both results in the absence of bone formation around the distraction area. Careful preservation of periosteum is essential to bone healing. Formation of bone with a large mineral content does not require bone marrow preservation, but there is an interaction effect on healing between bone marrow and periosteum. PMID- 9855239 TI - No adverse effects of clodronate on fracture healing in rats. AB - Clodronate was administered daily 28 days before and after an experimental tibial fracture in 35 male rats, and the effect on fracture healing and posttraumatic bone loss was studied. 5 groups were tested. The clodronate/clodronate group received clodronate in daily doses of 10 mg/kg body weight for 28 days before being subjected to a standardized fracture of the right tibia, and during the fracture healing period of 28 days. The clodronate/saline group received clodronate before fracture and saline during the healing period. The saline/clodronate group received saline before and clodronate after fracture. The saline/saline group received saline only, while the control group served as unfractured, untreated controls. After 28 days of fracture healing, the tibias were evaluated with dual energy x-ray absorptiometry, and tested mechanically in a 3-point ventral bending test. Bone mineral content and bone mineral density were approximately 30% higher in the groups receiving clodronate during the experiment, compared to the untreated groups. The weight and cross-sectional area of the fracture callus were equal in all groups. Whether clodronate was administered before the fracture, after the fracture or both, did not affect the bone mineral. Ultimate bending moment, energy absorption, stiffness and deflection were not significantly different between the groups. Our findings suggest that clodronate increases bone mineral both when given before and after a tibial shaft fracture, without affecting fracture healing at 28 days. PMID- 9855240 TI - Incorporation of cortical bone allografts and autografts in rats: expression patterns of mRNAs for the TGF-betas. AB - Healing of bone grafts is dependent on the rate of new bone formation. To understand better the regulation of new bone formation in the graft we have studied local production of TGF-beta1, 2 and 3, and of the small proteoglycans by determining their mRNA levels in a rat bone graft model. These mRNA levels were compared to the healing rates of autografts and allografts, as determined by histology, UV-microscopic evaluation of tetracycline-labeled new bone formation, microradiography and mechanical testing at 1, 2, 4 and 8 weeks of healing. Analyses showed that, analogous to slower bone formation in allografts, the induction of TGF-beta1 gene expression was slower than in allografts, when compared with autografts. A similar delay was seen in decorin gene expression. The results agree with the suggested role of TGF-beta1 in induction of type I collagen and osteonectin production. Our findings thus support the view that locally produced TGF-beta1 plays a role in normal graft incorporation, while local production of TGF-beta3, and particularly TGF-beta2, may be less important in this respect. PMID- 9855241 TI - Major fibrillar collagens and fibronectin in an experimental nonunion: an immunohistochemical study. AB - We studied in a rat pseudoarthrosis model the time sequence of expression and distribution of fibronectin and collagens I, II, III and V. Collagens and fibronectin were immunolocalized at the light microscopic level. The major difference from the normal healing pattern was the extension of collagen II and cartilage into the interfragmentary area and at the circumference of the periosteal callus, without any bridging chondral or bony elements in the fracture gap. The formation of a fibrous bond, consisting mostly of collagen III and fibronectin, was observed. This speaks in favor of the failure of the multipotentional mesenchymal stem cells to change the fracture-healing process towards fibroblast proliferation and the production of tissue unable to mineralize. The decisive zone for mineralization of the callus appeared to be the area of the hypertrophied chondrocytes near the periosteal ossification front. PMID- 9855242 TI - Ultrasound measurements of the newborn hip--comparison of two methods in 657 newborns. PMID- 9855243 TI - Human leukemic K562 cells treated with cytosine arabinoside: enhancement of erythroid differentiation by retinoic acid and retinol. AB - Human leukemia K562 cells can be induced to erythroid differentiation when treated with a variety of compounds, including hemin, cytosine arabinoside and 5 azacytidine. Following erythroid induction, K562 cells express at high level gamma-globin and accumulate both Hb Portland and Hb Gower 1. In this paper we determined whether a combination treatment of K562 cells with suboptimal concentrations of cytosine arabinoside and retinoids lead to full expression of differentiated functions. Cell growth kinetics studies, intracellular detection of hemoglobin by benzidine staining and hemoglobin analysis by cellulose acetate were performed. The results obtained show that (a) retinoic acid and retinol are not able to induce differentiation of K562 cells and (b) cytosine arabinoside induces differentiation only when used at 100-300 nmol/l concentrations. In addition, our data demonstrate that erythroid differentiation of K562 occurs when 40 micromol/l of retinoic acid or retinol are added together with 75 nmol/l cytosine arabinoside. PMID- 9855244 TI - Elevated IL-8 levels during sickle cell crisis. AB - The vaso-occlusive process (VOC) in sickle cell disease is of a complex nature. It involves intricate interactions between sickle red blood cells, endothelium and probably also leukocytes. As these interactions are regulated by cytokines, we analyzed the role of the potent neutrophil chemokine IL-8 by measuring serum levels in sickle cell patients during sickle cell crisis. These results were compared to nonsymptomatics and healthy controls. In patients having a vaso occlusive crisis both HbSS and HbSC patients showed significantly enhanced serum IL-8 levels compared to healthy controls. Several of these patients showed extremely elevated serum IL-8 levels which were independent of the crisis inducing factor. Furthermore, a sickle cell patient with VOC as a complication of rhGM-CSF treatment similarly showed high IL-8 serum levels at crisis onset. Nonsymptomatic sickle cell patients serum IL-8 levels were comparable to healthy controls. These results implicate a role for IL-8 at or during (the initiation of) sickle cell crisis. PMID- 9855245 TI - The use of intravenous intermediate dose melphalan and dexamethasone as induction treatment in the management of de novo multiple myeloma. AB - The variable absorption of melphalan from the gastrointestinal tract results in response rates between 40 and 60%. High dose melphalan increases response rates but at the cost of increased morbidity and mortality. We have investigated intravenous intermediate dose melphalan and dexamethasone in the treatment of patients presenting with de novo multiple myeloma with the object of reducing toxicity while preserving an improved response rate compared to oral melphalan and prednisolone. The results show that this treatment can be delivered safely on an outpatient basis in patients up to the age of 78 yr; 82% of patients achieved an objective response and 30% a complete haematological and clinical remission. Median overall survival for the whole group is 37 months. PMID- 9855246 TI - Serum levels of soluble vascular cell adhesion molecule-1 (sVCAM-1) are elevated in advanced stages of non-Hodgkin's lymphomas. AB - The serum levels of soluble vascular cell adhesion molecule-1 (sVCAM-1) were measured in 116 patients with non-Hodgkin's lymphomas (NHL) tested previously for soluble intercellular adhesion molecule-1 (sICAM-1). In contrast to Hodgkin's disease and chronic lymphocytic leukaemia, the sVCAM-1 levels in NHL patients were not significantly different from the levels of healthy controls (n=31). However, sVCAM-1 was elevated in advanced stage disease, i.e. stages III+IV. Elevated serum levels of sVCAM-1 were associated with significantly poorer disease-free (p = 0.024) and overall (p = 0.02) survival. sVCAM-1 correlated poorly with other known prognostic variables (LDH, sTK and beta2m) and with sICAM 1. None of the tested markers added prognostic information for disease-free survival independently of Ann Arbor stage and B-symptoms. The expression of VCAM 1 and ICAM-1 in tumour biopsies from 15 patients representing 7 different histologies were examined and compared with the serum levels of the soluble adhesion molecules. No correlation was found between the adhesion molecule expression by vascular endothelium and the corresponding serum levels. PMID- 9855247 TI - Platelet membrane lipid fluidity and intraplatelet calcium mobilization in type 2 diabetes mellitus. AB - The aim of the present study was to relate the impairments in calcium mobilization and/or release to the altered membrane dynamics in platelets from patients with type 2 diabetes mellitus. Higher expression of P-selectin (1.4 fold, NS) and the reduction in GPIb alpha expression (by 27.8+/-16.7%, p < 0.0002), as well as the increased fractions of platelet microparticles (p < 0.03), reflected more intensified platelet release reaction in diabetic platelets. Overall, diabetic platelets appeared more vulnerable to stimuli facilitating calcium mobilization (by 41%, p < 0.01) and less susceptible to preventive effects of the agents hampering calcium release from intraplatelet storage pools (by 38%, p < 0.01). Both the increased calcium mobilization from intraplatelet storage pools and higher levels of intracellular free calcium in the presence of procaine in diabetic platelets correlated with the reduced platelet membrane lipid fluidity (resp. pR < 0.03 and pR < 0.015). We conclude that the biophysical state of platelet membrane components in diabetes mellitus is the crucial determinant of platelet hyperfunction and probably contributes to the intensified calcium mobilization in diabetic platelets. The depressed preventive effects of procaine on platelet release reaction and calcium mobilization in diabetic platelets may result from the primary dislocations and/or distortions of membrane components caused by the diabetic state. PMID- 9855248 TI - Acute promyelocytic leukaemia: epidemiology and risk factors. A report of the GIMEMA Italian archive of adult acute leukaemia. GIMEMA Cooperative Group. AB - Acute promyelocytic leukaemia (APL) exhibits peculiar epidemiological, clinical, cytogenetic and molecular features, compared to the other acute myeloid leukaemias (AML). Data on epidemiology and occupational risk factors for APL desumed from the GIMEMA archive are reported and compared with those of the other AML. An exploratory case-case study was designed on AML patients from 56 haematology centres in Italy. Overall, 4296 patients older than 15 yr with a new diagnosis of acute leukaemia were recorded between July 1992 and July 1997. Of these, 335 were classified as APL, and 2894 as other AML. The median age of APL patients was 43 compared to 59 yr for the other AML (p < 0.00001). In order to identify peculiar risk factors for APL development, different parameters were compared in the 2 groups. After adjusting by age no significant differences were observed with regard to education, lifetime prevalence of cancer among siblings and previous diseases in the patient's history. Occupational exposure as a possible risk factor for APL showed no increased risk compared to other AML among farmers, builders and leather workers. A significant association was found in electricians (OR=4.4, 95% CI=2.0-9.7) and a weak association was found in wood workers (OR=3.2, 95% CI=0.8-10.8). The proportion of APL with respect to other AML was significantly higher in the north east of Italy compared to the rest of the country (OR=1.7, 95% CI=1.3-2.2). These data confirm the younger age of APL patients compared to the other AML. A possible role of electromagnetic fields is suggested by the higher risk of APL in electrical workers and in the more industrialized areas of the country. PMID- 9855249 TI - Platelet activation in the Dutch haemocytometry quality assessment programme: correlation between P-selectin expression and variation in platelet count. AB - Since 1990, our laboratory has prepared a set of 8 fresh whole blood samples for use in a countrywide quality assessment (QA) programme. The samples are intended as external controls for haemocytometry analysers. These samples are of 8 different haematocrit levels and each one is prepared from a single donor. About 210 laboratories participate in this QA programme. From the start of this programme large interlaboratory variations in platelet counts were encountered in some of the samples. This variability was much higher than would be expected and was independent of the platelet count of the samples. The main cause was thought to be formation of platelet aggregates. The aim of the present study was to find a parameter that can predict a high interlaboratory variation in the QA programme. Therefore we investigated the initial platelet activation status in the donors and the activation status of platelets in the prepared QA blood. As a marker for platelet activation P-selectin expression on the platelets was measured using flow cytometry. During 5 rounds of the QA programme we found a good correlation of r = 0.53 (p < 0.001) between P-selectin expression on platelets in the reconstituted QA blood and the interlaboratory platelet count variability. We conclude that P-selectin expression in the prepared QA blood is an important parameter to exclude samples that lead to high CVs of platelet counts in the QA programme. PMID- 9855250 TI - Flow cytometric identification of myeloid disorders by asynchronous expression of the CD14 and CD66 antigens. AB - Using a multiparameter flow cytometry assay enumerating cells positive for CD13, CD14 and CD66 antigens, we determined the asynchronous CD14/CD66 co-expression in unselected bone marrow and peripheral blood samples with suspected malignant blood disorders. CD14/CD66 co-expression > or = 5% were found in 131/691 bone marrow samples. Only 55 of these exhibited an identifiable population in 2 parameter flow cytometry histograms. Of the 55 samples 43 (78%) came from patients with myeloid disorders; e.g. 11 with myelodysplastic syndromes, 15 with chronic myeloproliferative disorders and 17 with acute myeloid leukaemia. Only one of these 17 cases was a de novo case, while 8 were secondary to another malignant haematological disease and 8 were from the period after cytoreductive therapy. Notably, CD14/CD66 co-expression patterns were related to disease categories; e.g. in chronic myelomonocytic leukaemia and acute myeloid leukaemia following a dysplastic phase the co-expression displayed two subsets in peripheral blood, low-avidity CD14 and low-avidity CD66, respectively. The latter disease category also exhibited these 2 subsets in bone marrow. In all other cases, the CD14/CD66 co-expression in bone marrow was heterogeneous. In conclusion, abnormal CD14/CD66 co-expression might be a valuable parameter in defining asynchronous myelopoiesis in malignant myeloid disorders, especially myeloproliferative disorders and secondary acute myeloid leukemias. PMID- 9855252 TI - Granulocyte colony-stimulating factor (G-CSF) may stimulate HIV-replication during cytostatic chemotherapy. PMID- 9855251 TI - Acute neurohumoral and cardiovascular effects of idarubicin in leukemia patients. AB - Idarubicin has been shown to have similar or superior antileukemic activity to daunorubicin with less cumulative cardiotoxicity. However, data of acute cardiovascular effects of idarubicin are scanty but may have clinical significance in predicting late cardiovascular complications. In the present study we evaluated prospectively acute neurohumoral and cardiovascular effects of idarubicin containing induction chemotherapy in 10 patients with newly diagnosed AML or MDS. Idarubicin was administered intravenously 12 mg/m2 on d 1, 3 and 5 as a part of the induction chemotherapy. Serial measurements of plasma atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were performed at baseline and the day following each idarubicin infusion. Echocardiography was performed to assess cardiac systolic and diastolic function. Signal averaged electrocardiography (ECG) was recorded to observe myocardial late potentials associated with possible myocardial injury. In addition, ambulatory ECG recording was performed to assess arrhythmias. Plasma concentrations of ANP increased from 18.2 +/- 1.5 pmol/l to 27.8 +/- 3.5 pmol/l (p = 0.011), to 30.2 +/- 3.0 pmol/l (p = 0.002) and to 40.8 +/- 6.0 pmol/l (p = 0.006) after the first, second and third doses of idarubicin, respectively. Similarly, plasma concentration of BNP increased from 6.2 +/- 1.9 to 9.0 +/- 1.8 pmol/l (p = 0.049) and 17.5 +/- 8.1 pmol/l (p = 0.203) after the first and third idarubicin infusion. Concomitantly, there was a trend towards an increase in left ventricular end diastolic diameter (LVEDD) (50.2 +/- 1.8 to 54.4 +/- 2.2 mm, p = 0.070). The increase in plasma BNP concentrations correlated significantly with the increase in LVEDD (r = 0.624; p = 0.002). No significant ECG changes or arrhythmias were associated with idarubicin infusions except in 1 patient who developed abnormal myocardial late potentials. Our results show that idarubicin causes acute neurohumoral activation associated with increased LVEDD indicating subclinical myocardial dysfunction. Whether these acute changes predict late clinical cardiomyopathy should be evaluated in prospective studies with larger number of patients and with higher cumulative anthracycline doses. PMID- 9855253 TI - Study on the measurement of the percentage of viable cells in frozen peripheral blood stem cell harvesting samples by an automated hematology analyzer. PMID- 9855254 TI - Office-based gastrointestinal endoscopy with intravenous sedation by the endoscopist. PMID- 9855255 TI - Cancer, inflammatory bowel disease, and the fecal stream. PMID- 9855256 TI - Esophageal rings, webs, and diverticula. AB - Rings, webs, and diverticula are among the most common anatomic anomalies of the esophagus. Although these structural lesions are often asymptomatic, patients can develop significant problems with dysphagia, regurgitation, and aspiration. This article discusses the epidemiology, pathogenesis, diagnosis, and therapy of esophageal rings, webs, and diverticula with emphasis on the clinical, diagnostic, and therapeutic strategies involved in caring for patients with these conditions. PMID- 9855257 TI - Endoscopic management of choledocholithiasis. AB - The first description of endoscopic sphincterotomy 25 years ago spearheaded the widespread use of endoscopic treatment of choledocholithiasis. It is largely accepted that common bile duct stone removal should be endoscopic rather than surgical in patients who have undergone previous cholecystectomy, in the high risk surgical patient when the gallbladder is still present, in patients with severe acute cholangitis, in selected patients with acute biliary pancreatitis, and in special circumstances for the average risk surgical patient with suspected choledocholithiasis before laparoscopic cholecystectomy. We have summarized a number of endoscopic techniques that are used in the management of bile duct stone disease. PMID- 9855258 TI - Outcomes research, practice guidelines, and disease management in clinical gastroenterology. AB - The delivery of health care is undergoing dramatic change. Often, patients have come to be viewed as customers and physicians as merely providers. The progressive rise in the cost of medical care has placed a premium on restraint. Unfortunately, quality has not received the same emphasis. As costs equalize among health care systems, however, quality of care will become the major differentiating feature among providers, including gastroenterologists. These changes have lead to what some term the third revolution in medical care, that is, "assessment and accountability". The field of outcomes research has emerged in response to these influences, providing the means to measure the relative effectiveness of different interventions to identify those that provide the most benefit for their cost. At the same time, outcomes research provides a monitoring system to ensure that quality is not sacrificed in the enthusiasm to reduce expenditures. These principles have implications in the practice of clinical gastroenterology. PMID- 9855259 TI - Chronic respiratory distress, dyspepsia, and diarrhea: What is the connection? PMID- 9855260 TI - Esophageal motility and intraesophageal pH patterns in patients with esophagitis and chronic nonsteroidal anti-inflammatory drug use. AB - Clinical reports suggest that nonsteroidal anti-inflammatory drugs (NSAIDs) use might induce esophageal symptoms and damage, including esophagitis, but experimental data are conflicting, and some indicate that NSAIDs improve mucosal damage. It is not known whether patients with endoscopically diagnosed esophagitis during NSAID use have different baseline reflux patterns from patients with reflux esophagitis and no NSAID use. Two groups of patients with and without chronic NSAID use and esophagitis were prospectively studied. Twenty four-hour pH monitoring and esophageal manometry were performed in all patients who were free of NSAID and any other drug use during tests. Esophageal motility and reflux patterns did not differ in patients with esophagitis regardless of the presence or absence of NSAID use. However, the subgroup of patients with grades II and III esophagitis and chronic NSAID use had a significantly greater lower esophageal sphincter (LES) pressure and a less severe intraesophageal pH profile than patients without NSAID use. In agreement with clinical reports, these results suggest that NSAID use may aggravate an otherwise milder acid-related esophageal disease in a subgroup of patients. PMID- 9855261 TI - Helicobacter pylori reinfection rate and duodenal ulcer recurrence in Korea. AB - In the short term, the eradication of Helicobacter pylori in patients with duodenal ulcer (DU) is deemed to be clearly effective; the long-term effectiveness apparently depends on the H. pylori reinfection rate. We conducted the present study to investigate the rates of H. pylori reinfection and DU recurrence in 45 patients previously cured of DU in whom H. pylori had been eradicated. These patients underwent gastroscopy and tests for H. pylori at least 1 year after eradication. In a control group comprising 31 patients with DU who were not treated with H. pylori eradication regimen, the DU recurrence rate was checked annually for 4 years. Twenty of 45 patients (44.4%) in whom the mean follow-up period was 3.5 years were again found to be H. pylori positive, and the reinfection rate was 12.8% per year. DU recurred in 8 of these 20 (40%) but not in any nonreinfected patients. In the control group, the DU recurrence rate was 61% within 1 year, 81% within 2 years, 84% within 3 years, and 90% within 4 years. The respective recurrence rates in the 45 patients in whom the bacteria had been eradicated were 0%, 4%, 13%, and 18%. The H. pylori reinfection rate was as high as 12.8% per year in Korea, but in that the DU recurrence rate is significantly lower (p < 0.01; odds ratio, 129.5) in the H. pylori-eradicated group than in the control group, the eradication of H. pylori in DU patients is effective over the long term (at least 4 years). PMID- 9855262 TI - A comparison of new rapid urease tests for detection of Helicobacter pylori: test characteristics and factors affecting positivity and time to positivity. AB - We assessed two new rapid urease tests, the Helicobacter Urease Test (HUT, Astra, Sweden) and the Polish test, for accuracy, reaction time, and the effect of biopsy site and bacterial density on test characteristics and time to positivity. A prospective study was conducted in two groups of patients: 64 consecutive patients undergoing upper endoscopy for dyspepsia and 61 consecutive patients with duodenal ulcer on upper endoscopy. In the first group, test accuracy, time to positivity, and possible associations with biopsy site and bacterial density were assessed between the tests. In the second group, the two new tests were compared with the CLOtest for time to positivity and effect of bacterial density on test outcome. The Polish and HUT test had similar specificities (97%), but the Polish test was more sensitive (90.3% vs. 80.7%). The Polish test was positive within 10 minutes in 55% of the positive patients compared with 10% for the HUT test. There was no association between bacterial density (by histologic count) and reaction time in either test or in the CLOtest in the second group. The Polish test was more accurate and had a quicker time interval to positivity than the HUT. There was no significant association between bacterial density and reaction time in any of the urease tests assessed, and the biopsy site did not affect test accuracy in the HUT test. PMID- 9855263 TI - No difference in seroprevalences of Helicobacter pylori infection between patients with pulmonary tuberculosis and those without. AB - A previous article reported a possible relationship between a history of tuberculosis and Helicobacter pylori infection. Epidemiologic similarities exist between the two infections: Mycobacterium tuberculosis and H. pylori are transmitted from person to person and the risk of acquiring them is elevated in underprivileged environment. This study was conducted to investigate the relationship between the two infections. Serum concentrations of anti-H. pylori IgG antibody were measured in 40 tuberculosis inpatients on antituberculosis chemotherapy for no more than 3 months (group I; 52.4 +/- 21.4 years of age), 43 tuberculosis inpatients on it for more than 3 months (group II; 57.3 +/- 16.3 years), and 60 nontuberculosis outpatients (control subjects; 55.9 +/- 16.7 years). H. pylori seropositivities were similar among control subjects (73.3%), group I (65%), and group II (69.8%). The difference in the antibody concentrations was significant between control subjects and group I (353.7 +/- 321.2 vs. 176.5 +/- 197.9 U/ml) but was not significant between control subjects and group II (353.7 +/- 321.2 vs. 229.9 +/- 249.5 U/ml). The seroprevalences may not be different between patients with pulmonary tuberculosis and those without, and antituberculosis therapy may not decrease the antibody concentrations. PMID- 9855264 TI - Gastric pathology in cholecystectomy patients: role of Helicobacter pylori and bile reflux. AB - The data regarding the role of Helicobacter pylori infection in patients with bile reflux are conflicting. Bile reflux is often observed after cholecystectomy. This study focuses on the role of H. pylori in gastric pathology of patients who had undergone cholecystectomy. Eighty-seven consecutive patients were included in the study. An upper gastrointestinal endoscopy was performed, and biopsy specimens were taken in the antrum, incisura angularis, and in the gastric body. The presence of bile reflux in gastric fluid at endoscopic examination was recorded. The overall H. pylori infection rate was 62%, with no difference between patients with (59.7%) and without (64%) endoscopic bile reflux (p = 0.67). The intestinal metaplasia rate in gastric mucosa was significantly higher in patients with both H. pylori and bile reflux than in patients without infection and bile reflux (36.4% vs. 5.6%, p = 0.02). Moreover, the mean number of years after cholecystectomy in patients with intestinal metaplasia was significantly higher than in those without metaplasia (21.1 +/- 7 vs. 11.5 +/- 8 years, p < 0.0001), whereas mean age did not differ significantly between groups (60.3 +/- 12 vs. 55.8 +/- 11 years, p = 0.14). Furthermore, we found four cases of gastric cancer, three with H. pylori infection. The mean number of years after cholecystectomy was significantly higher in patients with gastric cancer than in other patients (21.8 +/- 4 vs. 12.2 +/- 8 years, p = 0.009). This study found that H. pylori infection is frequent in cholecystectomized patients, also in the presence of endoscopic bile reflux. Bile reflux seems to act synergistically with H. pylori infection on gastric pathology. PMID- 9855265 TI - Liver function in cirrhotic patients in the euthyroid versus the hypothyroid state. AB - We recently showed that some decrease in thyroid hormone blood levels can effectively and significantly prevent the development of cirrhosis and fulminant hepatic failure and decrease portal pressure in three different rat models. This study was conducted to determine whether hypothyroidism has a beneficial effect over euthyroidism on patients with active liver cirrhosis of different etiologies. The medical files of hypothyroid patients with cirrhosis who were referred to the Tel-Aviv Medical Center between the years 1980 and 1995 were retrospectively evaluated. Of 3,528 patients with biopsy-proven cirrhosis and 4,738 hypothyroid patients who were identified, only 46 (25 female, 54%; mean age, 52.3 +/- 9.1) met the eligibility criteria. The patients suffered from cirrhosis (mean, 9.5 +/- 4.3 years; range, 4-23) and had hypothyroidism (mean, 12 +/- 6 years; range, 4-31). Most patients suffered from hypothyroidism of unknown etiology (85%), whereas the rest had hypothyroidism after surgical/iodine ablation of the gland. In the hypothyroid versus the euthyroid state, a significant negative correlation was found between thyroid-stimulating hormone blood levels and both functional and synthetic liver function tests (p < 0.001). A significant negative correlation was also found between thyroid-stimulating hormone blood levels and clinical deterioration manifested as bleeding varices, the development of ascites, and episodes of encephalopathy. We conclude that in patients with liver cirrhosis, the liver function in the hypothyroid state tend to be better than in the euthyroid state. A mild controlled decreased thyroid function may be beneficial for euthyroid cirrhotic patients. PMID- 9855266 TI - Extremely high levels of alkaline phosphatase in hospitalized patients. AB - We determined the diseases associated with extremely high levels of alkaline phosphatase in hospitalized patients. Computerized laboratory records of the Hospital of Saint Raphael identified all inpatients who had elevations of alkaline phosphatase above 1,000 U/l from April 1994 to September 1995. Thirty seven inpatients with alkaline phosphatase levels above 1,000 U/l were identified. Six had bone involvement from malignancy or Paget's disease and were eliminated from further analysis, and 31 patients were included in the study. Levels of alkaline phosphatase ranged from 1,014 to 3,360 U/l. Ten patients had sepsis as the cause of the elevated alkaline phosphatase. These included gram negative organisms, gram-positive organisms, and two patients with fungal sepsis. Seven of 10 patients with sepsis had an extremely high alkaline phosphatase level and a normal bilirubin, 3 of 10 patients with sepsis also had acquired immunodeficiency syndrome (AIDS). Eight patients had biliary obstruction, 7 with malignant obstruction and 1 with a common bile duct stone. Nine patients had AIDS. The cause of the elevated alkaline phosphatase in these included three with sepsis, three with mycobacterium avium intracellulare (MAI) infection, two with cytomegalovirus infection, and one with Dilantin toxicity. Three patients had diffuse liver metastases. Finally, four patients had benign intrahepatic disease, including one patient with liver hemangiomas, one patient with sarcoid hepatitis, one patient with lead toxicity, and one patient with drug-induced cholestasis. Extremely high elevations of alkaline phosphatase are most frequently seen in patients with sepsis, malignant obstruction, and AIDS. Patients with sepsis can have an extremely high alkaline phosphatase level and a normal bilirubin. A variety of other causes were also noted. PMID- 9855267 TI - Mycobacterium mucogenicum bacteremia in a patient with cirrhosis. PMID- 9855268 TI - Healing of cimetidine-resistant Menetrier's disease by eradication of Helicobacter pylori infection. AB - We report a case of Menetrier's disease associated with hypoproteinemia who was successfully treated by eradication of Helicobacter pylori infection. A 26-year old woman had diffuse giant gastric folds with proven protein leakage from the stomach. She received cimetidine, famotidine, and omeprazole for more than 3 months with little benefit. H. pylori infection was detected, and she was treated with clarithromycin (200 mg three times a day), lansoprazole (30 mg/day), and ecabet sodium (1.5 g twice a day) for 2 weeks. After starting the treatment, rapid increase of serum albumin and IgG levels were observed. The treatment resulted in eradication of H. pylori infection, and serum albumin and IgG levels returned to normal. Histologic specimens also showed the improvement of the irregularly elongated gastric pits with inflammatory infiltrates after the treatment. This case report suggests that eradication of H. pylori is recommended therapy for the immediate improvement of Menetrier's disease associated with hypoproteinemia. PMID- 9855269 TI - Granulomatous gastritis due to taeniasis. AB - Taeniasis is very common all over the world and invades the upper small bowel in humans. It is very unusual to see this parasite in the stomach. We report a case of gastric taeniasis that led to granulomatous gastritis. The elimination of the parasite resulted in complete disappearance of the granulomas in the corpus and antrum and in complete recovery of the patient. PMID- 9855270 TI - Iron deficiency anemia due to a Brunner's gland hamartoma. AB - Brunner's gland hamartomas are rare benign duodenal tumors often discovered incidentally during endoscopy or on an upper gastrointestinal series. These tumors arise mainly in the duodenal bulb and can present with gastrointestinal bleeding or symptoms of intestinal obstruction. When symptomatic, surgical or endoscopic removal can be safely performed and the prognosis is very good. We describe a 63-year-old man presenting with iron deficiency anemia due to a large Brunner's gland hamartoma and review the endoscopic, radiologic, surgical, and pathologic findings. PMID- 9855271 TI - Pancreatic cancer associated with granulocyte-colony stimulating factor production confirmed by immunohistochemistry. AB - We report an 83-year-old man with pancreatic body cancer of 4.5 cm in diameter. Peripheral leukocyte count was 15,700/microl and the serum concentration of granulocyte-colony stimulating factor (G-CSF) was 123 pg/ml (normal, 6.0-21.9 pg/ml) on admission. Furthermore, not only K-ras codon 12 (GGT --> GAT) but also p53 at codon 247 (CGG --> CCG) mutations were identified in the pancreatic juice aspirated endoscopically. We performed chemotherapy with two courses of 5 fluorouracil, pirarubicin hydrochloride, and mitomycin-C, resulting in no beneficial effect. After the second course the patient developed interstitial pneumonia, probably caused by anticancer drugs, and died 4 months after the tumor was detected. In the autopsy tissue, the tumor macroscopically occupied the pancreas body and was 7 x 6 x 5 cm in size. Histopathologic diagnosis of the tumor was poorly differentiated adenosquamous carcinoma. Immunohistochemical staining of the autopsy tissue showed that pancreatic cancer cells were positive for G-CSF. This is the first case report of G-CSF-positive pancreatic cancer confirmed by immunohistochemistry. PMID- 9855272 TI - Portal and mesenteric vein thrombosis in a patient heterozygous for a mutation (Arg506-->Gln) in the factor V gen (factor V Leiden). AB - In 30-50% of patients with portal thrombosis, no underlying etiology is found. The recent reports of new hereditary clotting defects are contributing to the understanding of this problem, but they only justify a small number of idiopathic cases. Instead, anticoagulant protein C resistance, caused by a mutation in the V factor gene, appears to be at least 10 times more common than any of the other known inherited deficiencies of anticoagulant proteins. In spite of that, extensive thrombosis of portomesenteric or hepatic venous circulation has been rarely described in this hereditary clotting defect. We report a typical case of familial and recidivant deep vein thrombosis in a young man heterozygous for the factor V Leiden mutation (Arg506-Gln), who developed an acute portal and mesenteric vein thrombosis. The patient was discharged with an oral anticoagulant treatment and remains asymptomatic 2 years later. In conclusion, the high prevalence of the factor V Leiden in young and aged patients with idiopathic vein thrombosis and the case here described makes it obligatory to consider this disorder in patients with portal and/or mesenteric vein thrombosis, especially in those without evident etiology. PMID- 9855273 TI - Spontaneous bacterial arthritis in a cirrhotic patient. AB - Septic arthritis is usually of hematogenous origin and is increasingly being reported in elderly patients, who often have underlying medical conditions such as diabetes or alcoholism. We report a 62-year-old patient with alcoholic liver disease who presented with Escherichia coli bacteremia and septic arthritis in a previously fractured ankle. There are scarce reports of infectious arthritis in cirrhotic patients, but this is the first report of arthritis after a primary enteric bacteremia. We believe that the patient described here developed E. coli bacteremia as a result of bacterial overgrowth and translocation related to alcoholic liver disease and cirrhosis. The resulting bacteremia resulted in the development of infection in the left ankle, which had preexisting disease and was thus vulnerable. This case provides further evidence for the mode of infection being bacteremia in cirrhotic patients. In patients with cirrhosis and fever, a high index of suspicion is required for joint infection as a potential cause of fever or deterioration in the cirrhotic's patient general condition. PMID- 9855274 TI - Treatment with low-dose interferon-alpha in Bulgarian HBeAg-negative patients with chronic hepatitis B. PMID- 9855275 TI - Safety of upper gastrointestinal endoscopy with intravenous sedation by the endoscopist at office: 17,963 examinations performed in a community center by two endoscopists over 17 years. PMID- 9855276 TI - Rectal leiomyosarcoma in a young male with a history of Wilms' tumor. PMID- 9855277 TI - Direct evidence of NO production in rat hippocampus and cortex using a new fluorescent indicator: DAF-2 DA. AB - The biological functions of nitric oxide in the neuronal system remain controversial. Using a novel fluorescence indicator, DAF-2 DA, for direct detection of NO, we examined both acute rat brain slices and organotypic culture of brain slices to ascertain NO production sites. The fluorescence intensity in the CA1 region of the hippocampus was augmented, especially after stimulation with NMDA, in acute brain slices. This NO production in the CA1 region was also confirmed in cultured hippocampus. This is the first direct evidence of NO production in the CA1 region. There were also fluorescent cells in the cerebral cortex after stimulation with NMDA. Imaging techniques using DAF-2 DA should be very useful for the clarification of neuronal NO functions. PMID- 9855278 TI - ERPs and chronometry of face recognition: following-up Seeck et al. and George et al. AB - Seeck et al. found that event-related potentials (ERPs) evoked by repeated and non-repeated face photographs differ as early as 50-70ms post-onset. They thus suggested that faces are recognized at these latencies, in contrast with current opinions in ERP literature. However, the similar latencies obtained by George et al. for stimuli not perceived as faces suggest that Seeck et al.'s differences could index repetition rather than face recognition per se. To address this issue, we used matched faces of known and unknown persons. We found the earliest differences between the ERPs to these faces between 76 and 130 ms. These results, which are consistent with other data, suggest that the differentiation of faces takes approximately 100 ms of processing time in humans. PMID- 9855279 TI - Attentional orienting and reorienting is indicated by human event-related brain potentials. AB - We investigated event-related potential indications for the orienting towards task-irrelevant, distracting aspects of stimulation and for the subsequent reorienting towards task-related aspects of stimulation. An identical experimental protocol was run in three conditions manipulating the task relevance of the sounds. As to be expected, distractors elicited the MMN (reflecting the brain's pre-attentive change detection) in each condition (even when the sounds were ignored) and subsequent N2b and P3 (reflecting orienting towards the distractor) when the sounds were attended. A late negativity was confined to a condition in which subjects discriminating long from short sounds were distracted by task-irrelevant frequency deviations. The 'reorienting negativity' (RON) probably reflects processes in the context of reorienting towards task-relevant aspects of stimulation following distraction. PMID- 9855280 TI - Event-related potentials during mental imagery. AB - Event-related potentials(ERPs) were measured while subjects were constructing mental images of a letter of the alphabet. Following presentation of an uppercase letter, subjects were asked to form a mental image of the same letter in lower case, and determine whether or not it had an ascending or descending stroke (ascender or descender). ERPs were measured in a passive condition and then compared with the ERPs recorded while the subjects performed the discrimination task. During the discrimination task, negative potentials were observed in left frontal, central and parietal areas around 220 ms after the stimulus onset. These early negative potentials were dominant in the left hemisphere and are probably related to working memory processes in mental image formation. PMID- 9855281 TI - Glucose plus insulin infusion improves cerebral outcome after asphyxial cardiac arrest. AB - Hyperglycemia before ischemia worsens cerebral outcome. The aim of this study was to determine the cerebral effects of giving glucose with or without insulin after asphyxial cardiac arrest. Rats underwent 8 min of asphyxial cardiac arrest. After arrest, Group 1 received NaCl; Group 2, insulin; Group 3, glucose; and Group 4, glucose plus insulin, all intravenously. Neurological deficit (ND) scores were 14+/-10%, 22+/-12%, 12+/-10% and 2+/-2% in Groups 1-4, respectively, 72 h after reperfusion. Overall histological damage (HD) scores were 4, 2, 3 and 1, respectively. Group 4 fared significantly better than group 1 on both scores. Glucose after asphyxial cardiac arrest in rats produces no increased brain damage while glucose plus insulin improves cerebral outcome. PMID- 9855282 TI - Substance P induces brief, localized increase in [Ca2+]i in dorsal horn neurons. AB - We determined the spatial and temporal dynamics of the increase in intracellular Ca2+ levels [Ca2+]i produced by substance P (SP) in dorsal horn neurons. A microinjection technique was used to apply minute amounts of SP to small areas of cultured neurons loaded with the Ca2+ indicator fura-2. Five successive applications of SP to the soma produced short-lasting (< 50 s) increases in [Ca2+]i that became gradually smaller, indicating receptor desensitization. Focal application of SP to a distal locus in a neurite produced a brief (12 s) increase in [Ca2+]i that travelled down the dendrite but did not spread into cell soma. Prolonged application of SP to these neurons caused the appearance of varicosities in their dendrites. PMID- 9855283 TI - Limbic ERPs predict verbal memory after left-sided hippocampectomy. AB - Surgical removal of the dominant medial temporal lobe regions runs a considerable risk of verbal memory deficits which may be compensated for postoperatively by corresponding regions in the non-dominant medial temporal lobe. We examined this possibility by recording event-related potentials (ERPs) to words from the medial temporal lobes of patients with left-sided temporal lobe epilepsy (TLE) undergoing presurgical evaluation. N400 amplitudes in the right anterior medial temporal lobe predicted the postoperative verbal recall performance of individual patients with surprising accuracy, indicating that intracranial recordings can be used to quantify the functional capacities of the right hemisphere that can compensate for the verbal memory deficits after loss of medial temporal lobe structures in the left hemisphere. PMID- 9855284 TI - Neural activity in SII modifies sensory evoked potentials in SI in awake rats. AB - The function of the projection from the secondary somatosensory cortex (SII) to the primary somatosensory cortex (SI) in rats was investigated by recording sensory evoked potentials (SEP) in SI during glutamate activation and lidocaine blockade of SII. In anesthetized animals, glutamate stimulation of SII decreased SEP latency and increased SEP amplitude, whereas no changes were evident during lidocaine blockade of SII. In awake animals, a second, later component of the SEP appeared. This second component was almost completely eliminated during lidocaine blockade of SII. We conclude that the projection from SII to SI in rats slightly facilitates the SEP response in anesthetized animals and is responsible for a major portion of the late component of the SEP in awake animals. PMID- 9855285 TI - A distinct low-level mechanism for interaural timing analysis in human hearing. AB - The detection of phase or timing differences, and amplitude differences between the two ears are cues for the spatial analysis of sound by humans. Previous physiological and anatomical studies of animals suggest that phase and amplitude differences between the ears may depend on different pathways, though human psychophysical studies suggest that interaural phase and amplitude differences between the two ears may be coded in the same way. Here we describe detailed psychophysical analysis of a subject with multiple sclerosis affecting the brain stem. He has a complete deficit in the detection of phase between the ears with preserved detection of interaural amplitude. The results prove that a distinct mechanism exists in humans for interaural phase detection. PMID- 9855286 TI - Pre-drug cues modulate morphine tolerance, striatal c-Fos, and AP-1 DNA binding. AB - To evaluate the molecular mechanisms that mediate the effect of learning on morphine tolerance in rats, we examined striatal c-Fos, and c-Jun protein expression, and AP-1 DNA binding. Morphine paired with a conditioned stimulus (CS) led to analgesic tolerance in the presence of the CS. Rats receiving morphine unpaired with the CS displayed significantly less tolerance than paired morphine animals. Striatal c-Fos protein levels and AP-1 DNA binding activity were increased in rats receiving paired morphine compared with rats that did not receive morphine but not in rats receiving morphine without the CS. No differences were found in c-Jun levels. These results suggest that Pavlovian conditioning may account, in part, for the molecular mechanisms associated with morphine tolerance. PMID- 9855287 TI - Elevated intracellular calcium levels in cerebellar granule neurons of weaver mice. AB - Weaver mice carry a mutation in the pore domain of the Girk2 (Kcnj6) gene. The mutation causes GIRK2 containing channels to lose ion selectivity and to become constitutively active. It is not known how this alteration in ion channel activity causes in cerebellar granule cells the defects in neurite extension, cell migration and induction of cell death that are characteristic of weaver mice. One possibility is that the mutation causes an inability to regulate intracellular calcium levels properly. We tested this hypothesis by measuring intracellular calcium levels in granule cells and Purkinje cells in slices from the cerebellum of weaver mice. We report here that weaver mice have increases in resting calcium levels in their granule cells, which may account for the multiple effects of the weaver mutation upon these cells. PMID- 9855288 TI - CB1 cannabinoid receptor antagonist-induced opiate withdrawal in morphine dependent rats. AB - Recent reports have provided evidence of a link between the endogenous brain cannabinoid system and the endogenous central opioid systems. Here we report that the selective CB1 receptor antagonist SR 141716A induced behavioral and endocrine alterations associated with opiate withdrawal in morphine-dependent animals in a dose-dependent manner and that naloxone induced an opiate withdrawal syndrome in animals made cannabinoid-dependent by repeated administration of the potent cannabinoid agonist HU-210. Additionally CB1 and mu-opioid receptor mRNAs were co localized in brain areas relevant for opiate withdrawal such as the nucleus accumbens, septum, dorsal striatum, the central amygdaloid nucleus and the habenular complex. These results suggest that CB1 cannabinoid receptors may play a role in the neuroadaptive processes associated with opiate dependence, and they lend further support for the hypothesis of a potential role of cannabinoid receptors in the neurobiological changes that culminate in drug addiction. PMID- 9855289 TI - BAPTA-AM blocks both voltage-gated and Ca2+-activated K+ currents in cultured bovine chromaffin cells. AB - The effects of the membrane permeant Ca2+ chelator BAPTA-AM on voltage-gated Na+, Ca2+, K+ (I(Na), I(Ca) I(K), respectively) and Ca2+-activated K+ (I(KCa)) currents in cultured bovine chromaffin cells were investigated using the whole cell patch-clamp technique. Superfusion with BAPTA-AM (50 microM) induced a rapid (< 60 s) and reversible block of both I(KCa) and I(K) (approximately 50%), without affecting either I(Ca) or I(Na). Preincubation with BAPTA-AM (50 microM, 30 min) or cell loading with the nonpermeable active form of BAPTA (10 mM in the pipette solution) permanently blocked I(KCa). BAPTA-AM superfusion (50 microM) also blocked I(K) (approximately 53%) after BAPTA-loading or BAPTA-AM preincubation. In conclusion, we show a fast and reversible block of I(KCa) and I(K) by BAPTA-AM, acting directly on K+ channels before it operates as a Ca2+ chelator, in cultured bovine chromaffin cells. PMID- 9855290 TI - Increased levels of TrkA in the regenerating retinal ganglion cells of fish. AB - Retinal ganglion cells of the fish have the spontaneous capacity to regenerate after nerve crush, a phenomenon known to be facilitated by nerve growth factor (NGF). We have studied the high-affinity NGF receptor TrkA, during the regeneration of the tench (Tinca tinca L.) optic nerve, using immunocytochemical techniques. TrkA-like immunoreactivity increased during the regeneration of the retinal ganglion cells. The increase is followed by a change in the subcellular distribution from perinuclear in control cells to cytoplasmic and perinuclear in regenerating ones. This increase was observed when antibodies against the extracellular domain of TrkA were used; no changes in TrkA-like immunoreactivity were observed with antibodies against the intracellular domain of TrkA. We thus conclude that modulation of TrkA is involved in the regeneration of fish retinal ganglion cells. PMID- 9855291 TI - Induction of neuropeptide Y expression in dorsomedial hypothalamus of diet induced obese mice. AB - Alterations of hypothalamic neuropeptide Y(NPY) and melanocortinergic functions in diet-induced obese (DIO) C57BL/6J mice were investigated by in situ hybridization. Compared with controls, the DIO mice displayed a profound induction (approximately 40-fold) of NPY expression in the dorsomedial (DMH) and ventromedial (VMH) hypothalamic nuclei, whereas the level of NPY mRNA in the arcuate nucleus (ARC) was reduced by 44%. The expression of pro-opiomelanocortin (POMC) and agouti-related protein was not significantly altered in the ARC of obese mice. Both excess body weight gain and altered hypothalamic NPY expression were reversible. We propose that the highly induced NPY expression in DMH and/or VMH may be a contributing etiological factor for the development of obesity and leptin resistance in the DIO mice. PMID- 9855292 TI - Anisotropic neural interaction in the primary auditory cortex of guinea pigs with sound stimulation. AB - Neural interaction in the primary auditory cortex of guinea pigs anesthetized with sodium pentobarbital was studied using a single line multi-electrode (4 x 1) aligned across and along the isofrequency band. Under the spontaneous condition, the neural interaction was isotropic; the amplitude of cross-correlogram peaks decreased as the electrode separation increased both across and along the isofrequency band. Under tone stimulation, the neural interaction was anisotropic; the amplitude of peaks was decreased rapidly beyond 400 microm across the isofrequency band, while it decreased little up to 700 microm along the isofrequency band. This anisotropic interaction was dependent on the stimulus intensity. PMID- 9855293 TI - Effects of stimulus intensity on signals from human somatosensory cortices. AB - We recorded somatosensory evoked magnetic fields (SEFs) to left median nerve electric stimulation from seven healthy subjects. The stimulus intensity was varied in three sessions: sensory stimuli evoked a clear tactile sensation without any movement, weak motor stimuli exceeded the motor threshold, and strong motor stimuli caused a vigorous movement. Responses were modelled with sources in the contralateral primary somatosensory cortex (SI), the contralateral and ipsilateral secondary somatosensory cortices (SIIs) and the contralateral posterior parietal cortex (PPC). The amplitude of the 20 ms response from the SI cortex and the subjective magnitude estimations followed the stimulus intensity whereas signals from the three other areas saturated already at the level of the motor threshold. The results implicate differential roles for various somatosensory cortices in intensity coding. PMID- 9855294 TI - Distinct short-term memory systems for sound content and sound localization. AB - Short-term memory for sound content and sound localization was investigated in normal subjects using the same/different comparison of two sound stimuli separated by an interval. Auditory or visual interference tasks requiring recognition or spatial judgements were introduced in the interval. Auditory interference tasks reduced memory for sound content and sound location in a specific way. Memory for sound content was significantly more reduced by auditory recognition than by auditory spatial interference task. Visual interference tasks reduced significantly memory for sound location but not for sound content. These results suggest that (i) short-term memory for sound content and that for sound location involve partially distinct processing; and (ii) auditory spatial functions are more closely linked to visual functions than auditory recognition. PMID- 9855295 TI - High-frequency repetitive transcranial magnetic stimulation delays rapid eye movement sleep. AB - Repetitive transcranial magnetic stimulation (rTMS) is a promising new treatment for patients with major depression. However, the mechanisms underlying the antidepressive action of rTMS are widely unclear. Rapid eye movement (REM) sleep has been shown to play an important role in the pathophysiology of depression. In the present study we demonstrate that rTMS delays the first REM sleep epoch on average by 17 min (102.6 +/-22.5 min vs 85.7+/-18.8 min; p < 0.02) and prolongs the nonREM-REM cycle length (109.1+/-11.4 min vs 101.8+/-13.2min, p< 0.012). These rTMS-induced changes in REM sleep variables correspond to findings observed after pharmacological and electroconvulsive treatment of depression. Therefore, it is likely that the capability of rTMS to affect circadian and ultradian biological rhythms contributes to its antidepressive action. PMID- 9855296 TI - Circadian rhythmicity in the locomotor activity of larval zebrafish. AB - The zebrafish (Danio rerio) may be useful for mutational analyses of vertebrate circadian clock mechanisms if efficient assays of circadian rhythmicity are available. Using an automated video image analysis system, we found robust circadian rhythms in the locomotor activity of larval (10- to 15-day-old) zebrafish maintained in constant conditions. Activity was rhythmic in > 95% of the animals tested. The timing of peak activity in constant conditions was determined by the prior light:dark cycle, with highest activity during the subjective day. The mean freerunning period of the activity rhythms was 25.6 h, and the within-experiment standard deviation of freerunning period ranged from 0.5 to 1.0 h. Therefore, it should be possible to detect mutations that lengthen or shorten the freerunning circadian period of zebrafish activity rhythms by 1-2 h. PMID- 9855297 TI - Magnolol protects cortical neuronal cells from chemical hypoxia in rats. AB - The protective effect of magnolol, a component of Magnolia officinalis, against hypoxia-induced cell injury in cortical neuron-astrocyte mixed cultures was examined. Exposure of the cells to chemical hypoxia (0.5 mM KCN) produced morphological changes in neurons but not in astrocytes. KCN induced dose- and time-dependent increases in release of LDH and decreases in viable cell number. Treatment with magnolol (10 and 100 microM) significantly reduced the KCN-induced LDH release in a concentration-dependent manner. A higher concentration (750 microM) magnolol was toxic. Nuclear condensation was not observed in KCN-treated cells, suggesting that chemical hypoxia-induced cell death was via necrosis, rather than via apoptosis. This is the first report demonstrating that magnolol protects neurons against chemical hypoxic damage or necrotic cell death in cortical neuron-astrocyte mixed cultures. PMID- 9855298 TI - Facilitation of acoustic responses of cartwheel neurons of the cat dorsal cochlear nucleus. AB - Responses to clicks were increased in cartwheel cells of the dorsal cochlear nucleus of cats after pairing presentations of the clicks with local iontophoretic delivery of glutamate. The cells were identified by bursting discharges, and were recorded intracellularly in vivo. The findings indicate that inhibitory interneurons such as cartwheel cells can participate in complex adaptive acoustic signal processing. Each cell displayed doublet discharges of > 800 Hz. In 70% of the cells, some of the doublet discharges reached rates > 1000 Hz. PMID- 9855299 TI - Neural correlates of cued recall with and without retrieval of source memory. AB - A source memory procedure was used to investigate whether event-related potentials (ERPs) reflect the retrieval of episodic information (recollection) on tests of word-stem cued recall. 'Cued recall ERP effects' for recalled items were found to differ only in their magnitudes, and not their scalp distributions, according to the accuracy of source memory. This finding strongly supports the hypothesis that cued recall ERP effects reflect quantitative variation in the amount or quality of information which can be retrieved about recently experienced events. Topographic analyses also revealed that the distribution of the cued recall ERP effects changed over time, reflecting the activity of at least two temporally and neuroanatomically dissociable neural populations contributing to recollection on this task. PMID- 9855300 TI - PKC and tyrosine kinase involvement in amyloid beta (25-35)-induced chemotaxis of microglia. AB - Microglia are activated by amyloid beta (Abeta) in vivo and in vitro, and Abeta activated microglia may be involved in the pathogenesis of Alzheimer's disease (AD). We investigated the mechanism of microglial chemotaxis induced by Abeta (25 35), an active fragment of Abeta. Abeta (25-35) 0.1 and 1 nM stimulated microglial chemotaxis. The protein kinase C (PKC) inhibitors chelerythrine (0.5 and 2 microM), calphostin C (1 microM) and staurospine (10 nM) significantly inhibited the microglial chemotaxis induced by Abeta (25-35) (1 nM). The chemotactic effect of Abeta (25-35) on microglia was desensitized by pretreatment of microglia with 1 ng/ml 12-O-tetradecanoylphorbol 13-acetate (TPA). Pretreatment of cells with Abeta (25-35) (1 nM) also desensitized the chemotactic effect by Abeta (25-35) (1 nM). The desensitization by TPA or Abeta (25-35) was inhibited when staurosporine was present in the pretreatment media. The tyrosine kinase inhibitor herbimycin A (0.1 and 1 microM) significantly inhibited the microglial chemotaxis induced by Abeta (25-35) (1 nM). Based on these observations, it seems likely that PKC and tyrosine kinase are involved in the Abeta-induced chemotaxis of microglia. PMID- 9855301 TI - Kainate-induced retina amacrine-like cell damage is mediated by AMPA receptors. AB - We investigated the effect of domoate, kainate and AMPA on 45Ca2+ uptake and on metabolic activity of cultured chick amacrine-like cells, as measured by reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Domoate and kainate stimulated 45Ca2+ uptake and decreased MTT reduction, in a LY 303070-sensitive manner. AMPA caused a small increase on 45Ca2+ uptake, but it was without effect on MTT reduction. AMPA reduced both the 45Ca2+ entry and neurotoxicity induced by kainate, and cyclothiazide enhanced both the 45Ca2+ entry and neurotoxicity induced by AMPA. The results indicate that the AMPA receptors are the non-NMDA glutamate receptors involved in excitotoxicity. PMID- 9855302 TI - Dexamethasone inhibits ischemia-induced transient reduction of neurotrophin-3 mRNA in rat hippocampal neurons. AB - Dexamethasone (DEX) increases the expression of neurotrophin-3 (NT-3) in normal rat hippocampal neurons, whereas transient forebrain ischemia reduces the NT-3 mRNA level. The effect of DEX on the expression of NT-3 mRNA in injured brain cells after ischemia has not been investigated, however. Using in situ hybridization and ribonuclease protection assay methods, we studied NT-3 mRNA expression in rats with and without DEX administration after transient forebrain ischemia. Without DEX treatment, NT-3 mRNA was down-regulated in the hippocampal neurons at 2, 4, 12 h and returned to basal levels 24 h following ischemia. With DEX treatment, however, NT-3 mRNA showed no change at 2, 4 and 12 h and increased 24 h after ischemia. The results indicate that DEX inhibits ischemia-induced NT-3 mRNA down-regulation during the first 12 h and up-regulates NT-3 mRNA 24 h after ischemia. DEX administration might be effective in influencing some of the pathophysiological effects of ischemia in the hippocampus. PMID- 9855303 TI - Muscarinic agonists exhibit functional dopamine antagonism in unilaterally 6-OHDA lesioned rats. AB - (5R,6R) 6-(3-propylthio-1,2,5-thiadiazol-4-yl)-1-azabicyclo[3.2.1]oc tane (PTAC) is a selective muscarinic ligand with high affinity for central muscarinic receptors, agonist mode of action at the muscarinic M2 and M4 receptor subtypes and substantially less or no affinity for central dopamine receptors. In the present study PTAC, as well as the muscarinic agonists oxotremorine, RS86 and pilocarpine, inhibited dopamine D1 and D2 receptor agonist induced contralateral rotation in unilaterally 6-OHDA lesioned rats. The dose of SKF 38393 used to induce contralateral rotation also caused an intense Fos protein immunoreactivity in the rat dorsolateral striatum on the lesioned site which was inhibited by PTAC indicating that the inhibition of rotation by PTAC was not due to non-specific peripheral side effects. PMID- 9855304 TI - Sensory neurons regenerate more dominantly than motoneurons during the initial stage of the regenerating process after peripheral axotomy. AB - This study was designed to determine whether sensory neurons or motoneurons were dominant during the earlier stage of the regeneration process after peripheral axotomy. After transection of the right sciatic nerves of rats, epineurial end neurorrhaphy was performed. At 5, 7 and 14 days postoperatively, the nerves were re-transected at the positive pinch site, and their proximal stumps were exposed to the retrograde neurotracer, Fluoro-Gold (F-G). Seventy-two hours later, the lumbar spinal cords and the L4 and L5 dorsal root ganglia (DRG) were harvested and evaluated. The incidence and the intensity of F-G labelling in DRG were significantly higher than in anterior horns (AH). These results demonstrated that sensory neurons were more dominant than motoneurons in nerve regeneration. PMID- 9855305 TI - Differential localization of GABA(B) receptors in the mouse retina. AB - The mRNA distribution of the two cloned GABA(B) receptor variants, GABA(B)R1a and -R1b, was analysed in the retina by non-radioactive in situ hybridization. GABA(B)R1a transcripts were found in the inner nuclear and ganglion cell layers, probably in horizontal, amacrine and ganglion cells, whereas GABA(B)R1b transcripts were detected in the ganglion cell layer only. Together with a recent immunohistochemical localization of GABA(B)R1 in the retina, this indicates a differential targeting of the receptor variants to pre- and postsynaptic sites with GABA(B)R1a and -R1b localized to axonal and dendritic compartments, respectively. In this way, inhibition of neurotransmitter release and slow postsynaptic inhibition could be provided by receptor variants derived from the same gene. PMID- 9855306 TI - Right hemisphere language in a neurologically normal dextral: a fMRI study. AB - Dextrals with right cerebral hemisphere dominance for language are rare. Eight neurologically intact dextrals underwent BOLD-fMRI while being presented auditory and visual words. Fortuitously, in one subject, right hemisphere activations with visually presented words were seen in the inferior frontal, premotor regions together with predominantly left cerebellar activation. These were a mirror image of activations obtained from the seven other dextrals. Also mirrored was temporal activation from auditory words which extended more posteriorly on the right side than the left. These results showing mirror organization of language were replicated in another scanning session and also by using a second word task. Although rare, mirrored organization of language can occur in normal dextrals without penalizing language function. PMID- 9855307 TI - Carnosine modulates zinc and copper effects on amino acid receptors and synaptic transmission. AB - Carnosine is a dipeptide which is highly concentrated in mammalian olfactory sensory neurons along with zinc and/or copper, and glutamate. Although carnosine has been proposed as a neurotransmitter or neuromodulator, no specific function for carnosine has been identified. We used whole-cell current- and voltage-clamp recording to examine the direct effects and neuromodulatory actions of carnosine on rat olfactory bulb neurons in primary culture. Carnosine did not evoke a membrane current or affect currents evoked by glutamate, GABA or glycine. Copper and zinc inhibited NMDA and GABA receptor-mediated currents and inhibited synaptic transmission. Carnosine prevented the actions of copper and reduced the effects of zinc. These results suggest that carnosine may indirectly influence neuronal excitability by modulating the effects of zinc and copper. PMID- 9855308 TI - Left prefrontal activation during episodic remembering: an event-related fMRI study. AB - The major current theory relating brain mechanisms in prefrontal cortex (PFC) to memory for discrete events (episodic memory) emphasizes the role of right PFC during retrieval. Using event-related fMRI, we found both right and left PFC activity during episodic remembering, but only the left PFC activity was related to the amount of episodic detail required at test. We suggest that right PFC subserves relatively simple, heuristic, cognitive processes and that left PFC is recruited for more reflectively demanding, systematic, processes. Episodic remembering often requires such systematic processes and, under those circumstances, recruits left, as well as right, PFC. PMID- 9855309 TI - Voltage-activated sodium currents in a cell line expressing a Cu,Zn superoxide dismutase typical of familial ALS. AB - The whole-cell configuration of the patch-clamp recording was used to study the voltage-dependent Na+ currents in a model system for the familial form of amyotrophic lateral sclerosis (ALS) associated with mutations in Cu,Zn superoxide dismutase. Here we report that the amplitude of voltage-gated Na+ currents is significantly reduced in cell lines expressing mutant Cu,Zn superoxide dismutase G93A when compared with the parental, untransfected cell line and to a cell line expressing the wild-type enzyme. This effect is associated with a shift toward positive values of the steady-state inactivation curve of the Na+ currents. These results indicate that expression of a Cu,Zn superoxide dismutase typical of patients affect with familial ALS influence the functionality of the voltage dependent Na+ channels; this effect may contribute to the pathogenesis of the disease. PMID- 9855310 TI - Selected pyrethroid insecticides stimulate glutamate uptake in brain synaptic vesicles. AB - We aimed to ascertain whether pyrethroid insecticides could influence the vesicular transport of the excitatory amino acid glutamate. The incubation of rat cortical synaptic vesicles with resmethrin and permethrin, consistently stimulated both ATP-dependent and -independent uptake of [3H]glutamate, while not evoking depletion of its vesicular content. Both processes were counteracted by valinomycin, a dissipator of the transmembrane potential gradient (deltapsi(sv)). Meanwhile, the vesicular influx of 36Cl- anions was impaired by pyrethroid concentrations which did not affect the ATP-dependent uptake of [14C]methylamine, as a marker for the proton gradient (deltapH). Thus, the stimulation of glutamate transport appeared to involve mainly the deltapsi(sv). A self-attenuating effect of selected pyrethroids on putatively enhanced excitatory transmission in severe intoxication is suggested. PMID- 9855311 TI - Protein kinase C inhibitors block generation of anoxia-induced long-term potentiation. AB - The aim of this study was to study the possible intracellular mechanisms underlying the anoxia-induced long-term potentiation (anoxic LTP) in the CA1 neurons of rat hippocampal slices using extra- and intracellular recording techniques. Superfusion of the hippocampal slices with the protein kinase C (PKC) inhibitors NPC-15437 (20 microM) or H-7 (20 microM) specifically prevented the induction of anoxic LTP. Moreover, the anoxic LTP was completely abolished in neurons intracellularly recorded with the selective PKC inhibitor PKCI 19-36 (50 microM). The specific cAMP-dependent protein kinase (PKA) inhibitor Rp-cyclic adenosine 3',5'-monophosphate (Rp-cAMPS, 25 microM) had no effect on the anoxic LTP. It is concluded that induction of anoxic LTP requires the activation of postsynaptic PKC. PMID- 9855312 TI - Redistribution of NMDA receptors in the cochlear nucleus following cochleotomy. AB - The major input to neurons of the cochlear nucleus comes from the glutamatergic cells of the spiral ganglion. We have studied the effect of unilateral destruction of the inner ear, including the spiral ganglion, with two antibodies against different types of NMDA receptor subunits, NMDAR1 and NMDAR2A/B, in the cochlear nucleus of the rat. Following cochleotomy, a dramatic redistribution of the receptor subunits was observed from a mostly perikaryal to a predominantly dendritic localization. Moreover, distinct changes in the composition of NMDA receptor complexes occurred. These effects were interpreted as compensatory responses to the massive loss of presynaptic release of the transmitter glutamate. PMID- 9855313 TI - Event-related brain activity associated with auditory pattern processing. AB - One of the basic properties of the auditory system is the ability to analyse complex temporal patterns. Here, we investigated the neural activity associated with auditory pattern processing using event-related brain potentials. Participants were presented with a continuously repeating sequence of four tones with rare changes in either the frequency or timing of one of the tones. Both frequency- and time-deviant sounds generated mismatch negativity (MMN) waves that peaked at midline central electrode sites and inverted in polarity at inferior temporal and occipital sites, consistent with generators in the supratemporal plane. The MMN scalp topography was similar for the frequency- and time-deviant stimuli, suggesting that both spectral and temporal relations among elements of an auditory pattern are encoded in a unified memory trace. PMID- 9855314 TI - Involuntary eye movements in response to first- and second-order motion. AB - We trained monkeys to maintain fixation while first- and second-order motion stimuli were displayed centrally in the visual field. Stimulus velocity, spatial frequency and contrast were varied to determine differences in patterns of involuntary eye movements elicited by random onset of stimulus motion. We observed different patterns of eye movement latency and velocity suggesting very early (< or = 100 ms) components of oculomotor activity are used to initiate smooth pursuit of object trajectory. Eye movement latency was insensitive to the complexity of stimulus motion, whereas second-order motion elicited faster eye movements than first-order motion. Instantaneous eye movement velocity might be related to the earliest stages of visual processing of component motion. PMID- 9855315 TI - Quetiapine, an atypical antipsychotic. AB - The discovery of antipsychotic agents in the 1950s revolutionized the treatment of schizophrenia. A large body of evidence supports the dopamine D2 receptor antagonist's efficacy in the treatment of psychotic symptoms. However, the advent of newer agents seems to point to a more complex interaction of neurotransmission in the pathophysiology of schizophrenia. In fact, a defining characteristic of atypical agents is a higher ratio of serotonin (5HT2) receptor blockade to D2 receptor blockade. Clozapine was the first atypical agent to be introduced; it was followed by risperidone, olanzapine, and now quetiapine, which is a dibenzothiazepine derivative structurally related to clozapine and olanzapine. PMID- 9855316 TI - Repaglinide, a novel, short-acting hypoglycemic agent for type 2 diabetes mellitus. AB - Repaglinide is a new, short-acting, insulin-releasing agent recently approved for the monotherapy of type 2 diabetes mellitus, and in combination with metformin in patients failing repaglinide or metformin monotherapy. Repaglinide appears to trigger insulin release by regulating adenosine triphosphate-sensitive potassium channels on the surface of pancreatic beta cells, which in turn affect calcium influx, the principal mediator of insulin release. Repaglinide mainly affects postprandial plasma glucose concentrations. It reduces glycosylated hemoglobin concentrations by 1-2% U. The agent's short duration of action may lessen the risk of long-lasting hypoglycemia and of down-regulation of beta cell sensitivity (the latter promoting secondary drug failure), although data are sparse in this regard. Its major adverse effect is hypoglycemia. Its role in the therapy of type 2 diabetes is unclear at present, vis-a-vis its use instead of or in combination with other antidiabetic agents other than metformin. The need for multiple daily doses, based on its brief duration of action, may be a barrier to compliance. PMID- 9855318 TI - Alteration in indinavir clearance during interleukin-2 infusions in patients infected with the human immunodeficiency virus. AB - STUDY OBJECTIVE: To evaluate the effect of interleukin-2 (IL-2) infusions on the pharmacokinetics of indinavir in patients infected with the human immunodeficiency virus. DESIGN: Observational, noncontrolled trial and prospective, open-label, nonrandomized, pharmacokinetic study. SETTING: Government research hospital. PATIENTS: Seventeen patients receiving indinavir 800 mg every 8 hours and a 5-day continuous infusion of recombinant IL-2. INTERVENTIONS: Observational study: trough indinavir concentrations were measured on day 1 and day 5 of IL-2 as part of a clinical trial. Prospective study: serial plasma samples were collected on days 1 and 5 of IL-2 to determine indinavir concentrations. Samples were also collected over the study period to determine IL 6 concentrations. The data were fit by a one-compartment model that allowed clearance to change based on IL-6 production and by standard noncompartmental equations. MEASUREMENTS AND MAIN RESULTS: The area under the curve of indinavir increased in eight of nine patients by a mean of 88% (range -29-215%) between days 1 and 5 of IL-2 infusion. Over this period, IL-6 concentrations also increased in all patients and indinavir clearance significantly decreased. Observational data in eight patients from the clinical trial showed significantly increased indinavir trough concentrations from 264+/-493 to 670+/-677 ng/ml in the presence of IL-2. CONCLUSION: Indinavir concentrations were altered during IL 2 infusions, possibly by induction of IL-6. Investigation into the effects of other proinflammatory cytokines is warranted. PMID- 9855317 TI - Pharmacokinetics of pyrazinamide under fasting conditions, with food, and with antacids. AB - STUDY OBJECTIVES: To determine intrasubject and intersubject variability in, and the effects of food and antacids on, the pharmacokinetics of pyrazinamide (PZA). DESIGN: Randomized, four-period, crossover phase I study. SUBJECTS: Fourteen healthy men and women volunteers. INTERVENTIONS: Subjects ingested single doses of PZA 30 mg/kg under fasting conditions twice, without a high-fat meal and with an aluminum-magnesium antacid. They also received standard dosages of isoniazid, rifampin, and ethambutol. MEASUREMENTS AND MAIN RESULTS: Serum was collected for 48 hours and assayed by gas chromatography with mass selective detector. Data were analyzed by noncompartmental methods and a compartmental analysis using nonparametric expectation maximization. Both fasting conditions produced similar results: mean PZA Cmax 53.4+/-10.4 microg/ml, Tmax 1.43+/-1.06 hours, and AUC(0 infinity) 673+/-79.7 microg x hr/ml. Fasting results are similar to those in previous reports. In the presence of antacids, subjects had a mean Cmax of 55.6+/ 9.0 microg/ml, Tmax of 1.43+/-1.23 hours, and AUC(0-infinity) of 628+/-88.4 microg x hr/ml. In the presence of the high-fat meal, mean Cmax was 45.6+/-9.44 pg/ml, Tmax 3.09+/-1.74 hours, and AUC(0-infinity) 687+/-116 microg x hr/ml. CONCLUSIONS: These small changes in Cmax, Tmax, and AUC(0-infinity) can be avoided by giving PZA on an empty stomach whenever possible. PMID- 9855319 TI - In unfractionated heparin dosing, the combination of patient age and estimated plasma volume predicts initial antifactor Xa activity better than patient weight alone. AB - STUDY OBJECTIVE: To determine whether patient factors other than body weight would better predict patients' initial antifactor Xa heparin activity (HA) after start of unfractionated heparin (UFH) therapy. DESIGN: Case series. SETTING: A 625-bed, adults-only, private, tertiary care teaching hospital. PATIENTS: Ninety two patients requiring UFH therapy. INTERVENTIONS: Patients received initial UFH bolus doses of 72-80 U/kg ideal weight and initial UFH infusions of 19.1-21.2 U/kg ideal weight. MEASUREMENTS AND MAIN RESULTS: Fifty-five percent of the first 6-hour HA measurements were supratherapeutic (> 0.7 U/ml antifactor Xa activity). Patient weight was inferior to a combination of age and estimated plasma volume in predicting initial HA. A predictive model including these two factors accounted for 38.5% of variation in first HA levels compared with 17.7% with actual weight alone. CONCLUSION: Weight-based UFH dosing may frequently result in nontherapeutic initial HA levels. Initial UFH dosing might be improved if protocols based on patient age and estimated plasma volume were developed. PMID- 9855320 TI - The pharmacokinetics and pharmacodynamics of fludarabine in rheumatoid arthritis. AB - STUDY OBJECTIVE: To describe the pharmacokinetics and pharmacodynamics of fludarabine in patients with rheumatoid arthritis (RA). DESIGN: Open-label, staggered trial conducted in conjunction with a phase I-II clinical trial. SETTING: Government research hospital. PATIENTS: Twenty-six patients with refractory RA. INTERVENTION: Fludarabine 20 or 30 mg/m2/day was administered as a 0.5-hour infusion for 3 consecutive days (1 cycle) for 6 months (1 cycle/mo). MEASUREMENTS AND MAIN RESULTS: Serial plasma samples were collected for pharmacokinetic analysis on day 2 of the first cycle of therapy. Relationships between pharmacokinetic parameters and hematologic and efficacy parameters were examined. The disposition of fludarabine was characterized by a two-compartment model. There were no differences in pharmacokinetics between the low- and high dose groups. The mean+/-SD total clearance, volume of distribution at steady state, and beta-half-life were 13.68+/-5.1 L/hour, 170.08+/-86.5 L, and 10.9+/ 3.1 hours, respectively. The volume of the peripheral compartment was approximately twice as large as the volume of the central compartment, indicating a significant amount of tissue distribution. No significant pharmacodynamic relationships were observed between pharmacokinetic parameters and hematologic and efficacy parameters. CONCLUSION: Fludarabine pharmacokinetics in patients with RA are characterized by an intermediate-length distribution phase (approximately 40 min), terminal half-life of 10.9 hours, and significant amount of tissue distribution. PMID- 9855321 TI - Pharmacokinetics of dexamethasone and valspodar, a P-glycoprotein (mdr1) modulator: implications for coadministration. AB - STUDY OBJECTIVE: To assess the potential for a drug-drug interaction between valspodar, a P-glycoprotein (mdrl) modulator used as a chemotherapy adjunct, and dexamethasone, widely included in oncology antiemetic regimens. DESIGN: Randomized, open-label, three-period crossover study. SETTING: Clinical pharmacology research center. SUBJECTS: Eighteen healthy men volunteers (age 25.8+/-3.5 yrs, weight 71.6+/-10.3 kg). INTERVENTIONS: Subjects received single fasting oral doses of valspodar 400 mg, dexamethasone 8 mg, and both drugs concomitantly with 2- to 3-week washout phases between administrations. MEASUREMENTS AND MAIN RESULTS: Lack of a pharmacokinetic drug-drug interaction with respect to valspodar was conclusively demonstrated for both Cmax,b (2.3+/ 0.4 vs 2.4+/-0.5 microg/ml) and AUCb (19.8+/-4.8 vs 19.6+/-4.9 microg x hr/ml) inasmuch as bioequivalence criteria were satisfied when comparing administration alone with coadministration, respectively. Although no changes in the rate of dexamethasone absorption were noted on coadministration with valspodar (Cmax 88+/ 23 vs 91+/-20 ng/ml), overall exposure was significantly increased by 24% on average (AUC 400+/-87 vs 494+/-90 ng x hr/ml). Regression analysis of valspodar Cmax,b and AUCb during coadministration versus the extent of the interaction (percentage increase in dexamethasone AUC) did not reveal a concentration-effect relationship (p=0.7299 and 0.9718, respectively). CONCLUSION: Given dexamethasone's wide therapeutic index and the short duration of coadministration foreseen for these drugs in a clinical setting (maximum 1 wk/chemotherapy cycle), the 24% increase in dexamethasone's AUC is unlikely to be relevant. Thus no alterations in valspodar or dexamethasone dosages appear warranted when the two drugs are coadministered. Multiple-dose experience in patients would be desirable to confirm these conclusions. PMID- 9855322 TI - Lack of effect of olanzapine on the pharmacokinetics of a single aminophylline dose in healthy men. AB - STUDY OBJECTIVE: To test whether olanzapine, an atypical antipsychotic, is an inhibitor of cytochrome P450 (CYP) 1A2 activity, we conducted a drug interaction study with theophylline, a known CYP1A2 substrate. DESIGN: Two-way, randomized, crossover study. SETTING: Clinical research laboratory. SUBJECTS: Nineteen healthy males (16 smokers, 3 nonsmokers). INTERVENTIONS: Because the a priori expectation was no effect of olanzapine on theophylline pharmacokinetics, a parallel study using cimetidine was included as a positive control. In group 1, 12 healthy subjects received a 30-minute intravenous infusion of aminophylline 350 mg after 9 consecutive days of either olanzapine or placebo. In group 2, seven healthy subjects received a similar aminophylline infusion after 9 consecutive days of either cimetidine or placebo. MEASUREMENTS AND MAIN RESULTS: Concentrations of theophylline and its metabolites in serum and urine were measured for 24 and 72 hours, respectively. Plasma concentrations of olanzapine and its metabolites were measured for 24 hours after the next to last dose and 168 hours after the last olanzapine dose. Olanzapine did not affect theophylline pharmacokinetics. However, cimetidine significantly decreased theophylline clearance and the corresponding formation of its metabolites. Urinary excretion of theophylline and its metabolites was unaffected by olanzapine but was reduced significantly by cimetidine. Steady-state concentrations of olanzapine (15.3 ng/ml), 10-N-glucuronide (4.9 ng/ml), and 4'-N-desmethyl olanzapine (2.5 ng/ml) were observed after olanzapine 10 mg once/day and were unaffected by coadministration of theophylline. CONCLUSION: As predicted by in vitro studies, steady-state concentrations of olanzapine and its metabolites did not affect theophylline pharmacokinetics and should not affect the pharmacokinetics of other agents metabolized by the CYP1A2 isozyme. PMID- 9855323 TI - Acute effects of prechemotherapy filgrastim administration on late hematopoietic progenitor cells. AB - Administration of filgrastim influences the proliferative kinetics of myeloid progenitor cells. Even after it is discontinued, increased levels of cycling of granulocyte precursors are sustained for approximately 4 days. Beginning chemotherapy during this period of enhanced marrow activity can cause great damage to late, and possibly early, progenitor cell pools. Peripheral blood samples were collected from two research study patients who were prospectively randomized to receive filgrastim by different schedules after chemotherapy. The mononuclear cell fraction was analyzed by clonogenic progenitor cell assay and flow cytometry. Ex vivo and clinical findings were correlated with filgrastim and chemotherapy administration times. Although quantitative recovery of circulating neutrophils occurred, a substantial decrease in peripheral blood progenitor cells was observed when chemotherapy was started 72 hours after cessation of filgrastim therapy. Neutrophil recovery alone is not a precise index of short-term marrow granulocyte progenitor status. Starting chemotherapy within 72 hours of filgrastim therapy is biologically, and possibly, clinically relevant. PMID- 9855324 TI - A multicenter, investigator-blinded, randomized comparison of oral levofloxacin and oral clarithromycin in the treatment of acute bacterial sinusitis. AB - A multicenter, investigator-blinded, randomized, parallel-group study was conducted to compare oral levofloxacin 500 mg once/day for 14 days with clarithromycin 500 mg twice/day for 14 days in the treatment of acute bacterial sinusitis. Of 216 adult outpatients randomized to treatment, 190 were evaluable for efficacy. The primary efficacy measure was clinical response, based on resolution of signs and symptoms 2-5 days after therapy. A secondary efficacy measure was relapse rate 1 month after therapy. Among evaluable patients, clinical success rates (cured or improved) were 96.0% and 93.3% for levofloxacin (L) and clarithromycin (C), respectively (95% CI -9.2%, 3.7%). The confidence interval (CI) for treatment difference (C-L) included zero and its upper limit was less than 15%, indicating that levofloxacin was as effective as clarithromycin. In all, 4.1% of patients receiving levofloxacin and 7.2% receiving clarithromycin had a relapse of symptoms 1 month after therapy (95% CI 12.2%, 3.2%). Long-term success (initial success, absence of relapse at 1 month, no further antibacterial therapy 2-5 days after therapy) was 79.2% in the levofloxacin group and 76.4% in the clarithromycin group (95% CI -14.7%, 9.0%). Based on investigator-assessed treatment-emergent adverse events, overall tolerability of the drugs was similar, except for a higher frequency of taste perversion and diarrhea in the clarithromycin group. Levofloxacin had an advantage over clarithromycin based on two quality-of-life (QOL) parameters: number of times taking other drugs for targeted medical conditions and mean total cost of these drugs. No statistical significance was found in other QOL variables. These findings suggest that the efficacy and tolerability of levofloxacin 500 mg once/day are comparable with those of clarithromycin 500 mg twice/day in the treatment of acute bacterial sinusitis. PMID- 9855325 TI - Retrospective and prospective analyses of the treatment of overanticoagulated patients. AB - STUDY OBJECTIVE: To compare withholding warfarin therapy with low-dose (2.5 mg) oral vitamin K therapy in excessively anticoagulated patients without bleeding complications. DESIGN: Prospective and retrospective studies. SETTING: Anticoagulation clinic at a Veterans Affairs institution. PATIENTS: Twenty-eight men were matched according to initial international normalized ratio (INR) and INR goal ranges. INTERVENTIONS: The retrospective arm of the study consisted of chart reviews of overanticoagulated patients whose warfarin doses were held until therapeutic INR values were reached. The prospective arm included overanticoagulated patients who were administered a single 2.5-mg dose of oral vitamin K. MEASUREMENTS AND MAIN RESULTS: Mean days to therapeutic INR values were 2.3+/-0.6 and 1.4+/-0.6 (p=0.001), and mean reduction in INR 1 day after treatment intervention was 1.32+/-0.79 and 3.46+/-1.31 (p<001) U for the withholding and vitamin K groups, respectively. CONCLUSION: Compared with withholding the warfarin dose, administration of 2.5 mg of oral vitamin K to excessively anticoagulated patients receiving warfarin significantly reduced the time required to reach a therapeutic INR as well as final INR. PMID- 9855326 TI - Cytochrome P450 3A4 activity in premenopausal and postmenopausal women, based on 6-beta-hydroxycortisol:cortisol ratios. AB - STUDY OBJECTIVE: To characterize cytochrome P450 (CYP) 3A4 activity in premenopausal and postmenopausal women by evaluating the urinary 6-beta hydroxycortisol:cortisol ratio. DESIGN: Prospective study SUBJECTS: Thirteen premenopausal and 13 postmenopausal women who were healthy and not receiving drugs known to affect CYP3A4 activity INTERVENTIONS: Beginning on day 2 of menses, premenopausal women collected first morning urine samples every other day for a complete menstrual cycle. Postmenopausal women collected first morning urine every other day for 28 days. MEASUREMENTS AND MAIN RESULTS: Mean weekly 6 beta-hydroxycortisol:cortisol ratios did not differ during the phase (week) of the menstrual cycle. Daily ratios did not differ in postmenopausal women. No difference between premenopausal and postmenopausal women was found on comparing overall median ratios. CONCLUSION: Cytochrome P450 3A4 activity as measured by 6 beta-hydroxy cortisol:cortisol ratio did not differ by week of menstrual cycle, suggesting no menstrual cycle-related changes. Menopause does not appear to be associated with differences in CYP3A4 activity, compared with premenopause. PMID- 9855327 TI - Antiepileptics and blood dyscrasias: a cohort study. AB - We conducted a cohort study to investigate the frequency of serious blood dyscrasias in patients age 10-74 years, taking antiepileptic drugs between January 1, 1990, and October 31, 1994. Main outcome measures were validated diagnoses of neutropenia, agranulocytosis, hemolytic anemia, thrombocytopenia, bicytopenia, pancytopenia, or aplastic anemia. A total of 29,357 recipients of antiepileptic therapy received 684,706 prescriptions. Among them there were 21 cases of serious blood dyscrasia of which only 18 could be considered to have a temporal relationship to drug use. Seventeen cases occurred in current users of carbamazepine, phenobarbital, phenytoin or valproate, and 7 in patients taking two or more drugs. Twenty of the 21 patients recovered. The overall rate of blood dyscrasias was 3-4/100,000 prescriptions. The rate in those age less than 60 years was 2.0 (range 0.9-3.6)/100,000 prescriptions compared with 4.0 (range 1.6 8.2) for those age 60 or older. The overall rate of neutropenia was 1.2 (0.5 2.3)/100,000 prescriptions, compared with 0.9 (0.3-1.9) for thrombocytopenia and 0.4 (0.1-1.3) for hemolytic anemia. Rates did not differ among the four drugs. Serious blood dyscrasias are rare in patients taking antiepileptic agents. PMID- 9855328 TI - Characteristics associated with ability to prevent adverse drug reactions in hospitalized patients. AB - We conducted a retrospective analysis to identify characteristics of preventable adverse drug reactions (ADRs). We reviewed reports on 612 ADRs occurring in hospitalized patients over 4 years, identified by the hospital's spontaneous ADR reporting program, and classified the events as potentially preventable or not preventable. Characteristics related to ADR preventability in the univariate analysis were the patient's clinical service, organ system involved in the ADR, class of drug causing the ADR, relationship to dosage, type of ADR, and probability that the reaction was due to the drug. Among these, relationship to dosage (p<0.001) and type of ADR (p<0.001) appeared to be most strongly related to preventability. In a multivariate analysis, preventable ADRs were associated with dosing (OR 3.82, 95% CI 2.42-6.03) and previous allergy to the drug (OR 3.46, 95% CI 1.01-11.88). An ADR that was classified as an allergic (OR 0.50, 95% CI 0.27-0.94) or idiosyncratic reaction (OR 0.44, 95% CI 0.28-0.71) was unlikely to be considered preventable. Preventable ADRs in hospitalized patients are likely to be dosage related or to occur among patients allergic to the specific agent. PMID- 9855329 TI - Safety of GM-CSF in patients with AIDS: a review of the literature. AB - We performed a literature search for all clinical studies reporting outcomes in patients with the acquired immunodeficiency syndrome (AIDS) receiving granulocyte macrophage colony-stimulating factor (GM-CSF) for any indication. Safety outcomes included human immunodeficiency virus replication, immune status, and frequency of opportunistic infections and neoplasms. Data were synthesized qualitatively. We identified 22 studies (274 patients): 12 addressed AIDS neutropenia, 8 AIDS cancer therapy, and 2 opportunistic infections. Viral burden was assessed by serum p24Ag in 15 studies. Nine reported no change in levels, three net decreases, and three net increases. All studies showing net increases involved patients receiving GM-CSF without a concurrent antiretroviral. The CD4 counts were unchanged in 5 studies, increased in 3, and not reported in 14. The incidence of neoplasms or new opportunistic infections was low. The literature suggests no increased risk of viral replication or clinical deterioration in patients with AIDS who take GM-CSF concurrently with zidovudine. PMID- 9855330 TI - Concomitant therapy with anxiolytics or hypnotics and maintenance of initial SSRI therapy. AB - We conducted a retrospective analysis to evaluate the relationship between anxiolytic or hypnotic therapy and maintenance of therapy with selective serotonin reuptake inhibitors (SSRIs). Subjects were 654 patients who received anxiolytics or hypnotics early in SSRI therapy (study group ) and 15,172 patients who did not (controls). Maintenance of SSRI therapy was evaluated during the 6 months after start of therapy and included days of initial SSRI therapy and rates of discontinuation, defined as a break of more than 30 days. Rates of discontinuation in study and control groups (84% and 77%, p=0.001) and average days of initial SSRI therapy (77 and 94 days, p<0.0001) were statistically different. Thus patients receiving anxiolytic or hypnotics in the first 60 days of therapy were less likely to continue initial SSRI therapy. PMID- 9855331 TI - Pharmacokinetic drug interactions of vinca alkaloids: summary of case reports. AB - Involvement of the cytochrome P450 (CYP) 3A subfamily in the metabolism of vincristine is well established. However, information is limited regarding vincristine's drug interaction profile. All the substrates and inhibitors of CYP3A4 such as the azole antifungals (itraconazole, ketoconazole), cyclosporine, isoniazid, and nifedipine have very high propensity to interfere with vincristine metabolism. The proposed mechanism is most likely attributed to either inhibition of 3A4 enzymes or blockade of P-glycoprotein pumps. These interactions are clinically significant and can lead to severe vincristine toxicity if not detected. Although case reports discussed here exclusively involve vincristine, it is important to assume that all vinca alkaloids interact in the same manner until proved otherwise, because they share similar metabolism pathways. PMID- 9855333 TI - Safety issues in the pharmacologic management of chronic pain in the elderly. AB - Chronic pain is commonly encountered in elderly patients. About 20-50% of community-dwelling elderly experience it, and 45-80% of nursing home residents may be affected. Selection of pharmacologic therapy for the management of chronic pain must take into consideration the increased potential for adverse effects in this population. Major classes of drugs used to treat chronic pain (nonsteroidal antiinflammatory drugs, opioids, antidepressants) have adverse effects that occur more frequently in elderly than in younger patients. Given the often prolonged duration of therapy, optimal management requires minimizing the risk of adverse reactions. PMID- 9855332 TI - Withdrawal after narcotic therapy: a survey of neonatal and pediatric clinicians. AB - Pharmacists at the 1995 American College of Clinical Pharmacy Pediatric Practice and Research Network meeting volunteered to act as coordinators at their sites and survey pediatric and neonatal nurses, pharmacists, and physicians regarding dependency in neonatal and pediatric patients after therapeutic administration of narcotics. Thirteen (60%) of 21 coordinators returned 244 surveys. Primary symptoms of withdrawal reported by clinicians were agitation (100%), irritability (100%), inconsolability (100%), crying (99%), tremors (98%), high heart rate (98%), fidgets (98%), high blood pressure (97%), less sleep (96%), and sweating (94%). Most clinicians considered narcotic withdrawal to be a problem (74%) that should be treated (87%). A dependency scale is being developed and will include symptoms reported by more than 75% of respondents. PMID- 9855334 TI - Effects of ACE inhibitor therapy on quality of life in patients with heart failure. AB - In addition to effects on survival, hemodynamics, and exercise capacity, quality of life has become an important outcome of therapy for chronic heart failure. A large clinical trial of the angiotensin-converting enzyme (ACE) inhibitor enalapril reports that certain domains of health-related quality of life (HRQL) have a long-term impact on survival in patients with reduced left ventricular systolic function, regardless of symptoms of heart failure at diagnosis. Results of large-scale clinical trials that measured the impact of several different ACE inhibitors on quality of life in these patients suggest benefits of the drugs, but data are confounded by a definite placebo effect. Studies are further confounded by high noncompletion rates for sicker patients, particularly beyond 1 year. Because measurement tools varied and different quality of life domains were evaluated, direct comparison of studies is problematic. Typically, HRQL measurements in patients receiving ACE inhibitors showed small improvement or did not differ significantly from those in placebo-treated patients with long-term follow-up, although short-term trials (< 6 mo) showed some benefit. Moreover, multicenter trials such as SOLVD, V-HeFT II, and ramipril studies indicated that ACE inhibitors do not compromise and may actually improve certain components of quality of life in a large number of patients with chronic heart failure secondary to reduced left ventricular systolic function while having favorable effects on survival, exercise capacity, hemodynamics, or symptoms. PMID- 9855335 TI - Cost-effectiveness of enoxaparin versus low-dose heparin for prophylaxis against venous thrombosis after major trauma. AB - We attempted to determine health and economic outcomes from the perspective of an integrated health system of administering enoxaparin 30 mg twice/day versus heparin 5000 U twice/day for prophylaxis against venous thrombosis after major trauma. A decision-analytic model was developed from best literature evidence, institutional data, and expert opinion. We assumed that 40% of proximal deep vein thromboses (DVTs) and 5% of distal DVTs are diagnosed and confirmed with initial or repeat duplex scanning; 50% of undiagnosed proximal DVTs result in pulmonary embolism; 2% and 1% of undiagnosed proximal DVTs will lead to readmission for DVT and pulmonary embolism, respectively, and pulmonary embolism-related mortality rates range from 8-30%. Length of hospital stay data and 1996 institutional drug use and acquisition cost data were used to estimate the cost of enoxaparin and heparin therapy. Diagnosis and treatment costs for DVT and pulmonary embolism were derived from institutional charge data using cost:charge ratios. A second analysis of patients with lower extremity fractures was completed. One-way and multiway sensitivity analyses were performed. For 1000 mixed trauma patients receiving enoxaparin versus heparin, our model showed that 62.2 (95% CI -113 to 12) DVTs or pulmonary emboli would be avoided, resulting in 67.6 (8 to 130) life years saved at a net cost increase of $104,764 (-$329,300 to $159,600). Enoxaparin versus heparin resulted in a cost of $1684 (-$3600 to $9800) for each DVT or pulmonary embolus avoided and a discounted cost/life-year saved of $2303 ( $8100 to $19,000). For 1000 patients with lower extremity fractures, enoxaparin versus heparin resulted in a cost of $751 (-$4200 to $3300) for each DVT or pulmonary embolus avoided and a discounted cost/life-year saved of $1017 ( $10,200 to $6300). Although enoxaparin increases overall health care costs, it is associated with a cost/additional life-year saved of only $2300, which is generally lower than the commonly used hurdle rate of $30,000/life-year saved. The cost-effectiveness ratio is more favorable in patients with lower extremity fractures than in the general mixed trauma population. PMID- 9855337 TI - Administration of filgrastim in two patients with drug-induced agranulocytosis. AB - Two patients experienced agranulocytosis associated with either procainamide or trimethoprim-sulfamethoxazole. Both were treated with filgrastim in an attempt to decrease the duration of agranulocytosis. The first patient received 12 days and the second patient 4 days of filgrastim before white blood cell counts recovered. Published reports both support and refute the efficacy of filgrastim in this setting. The agent may be beneficial in certain cases of this disorder, but further investigation is necessary to determine if it has a definitive role. PMID- 9855336 TI - The pharmacokinetics of intravenous estradiol--a preliminary study. AB - Estradiol USP was extemporaneously compounded for intravenous administration. Eight postmenopausal women were randomized to receive one of four estradiol dosages. Serum estradiol concentrations were determined at frequent intervals after single bolus dosing. The concentration-time profile was stripped and fit, and pharmacokinetic values were generated. Approximate dosage proportionality was seen with area under the curve, the terminal half-life was 27.45+/-5.65 minutes, and volume of distribution was very low (0.082+/-0.015 L/kg). Estradiol was well tolerated by all study participants. PMID- 9855338 TI - Ticlopidine-induced thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura (TTP) occurs in association with a wide variety of disorders including infections, connective tissue diseases, and solid organ tumors. It also may coincide with administration of drugs such as mitomycin, metronidazole, oral contraceptives, cyclosporine, and many others. We report the occurrence of TTP in a patient shortly after the initiation of ticlopidine. PMID- 9855340 TI - Enhanced metabolism of levonorgestrel during phenobarbital treatment and resultant pregnancy. AB - Levonorgestrel implants (Norplant) are an alternative to oral contraceptives and medroxyprogesterone acetate intramuscular injections. An interaction may exist between levonorgestrel and agents that induce the hepatic microsomal enzyme system. A 21-year-old woman with a history of a seizure disorder, treated with phenobarbital, who received levonorgestrel implants became pregnant. After a normal delivery, she took oral contraceptives concomitantly with phenobarbital. Although she was educated about the importance of a backup method of contraception, the woman again became pregnant and delivered twins. A recent national survey of neurologists and obstetricians was conducted evaluating prescriber knowledge of interactions between oral contraceptives and anticonvulsants. Only 4% of neurologists and zero percent of obstetricians knew all the interactions between the six most commonly prescribed anticonvulsants and oral contraceptives. This case supports the importance of continued patient and prescriber education regarding the possibility of drug-drug interactions in women taking anticonvulsants and hormonal contraceptives. PMID- 9855339 TI - Interaction between tacrolimus and nefazodone in a stable renal transplant recipient. AB - Tacrolimus (FK-506) is an important immunosuppressive agent most often given for maintenance immunosuppression to prevent acute cellular organ rejection. A 57 year-old woman with end-stage renal disease presumed secondary to chronic glomerulonephritis underwent a living related renal allograft transplantation. She tolerated the surgery well and was discharged on postoperative day 5. She was stabilized with prednisone, azathioprine, and tacrolimus. Two years after transplantation, nefazodone 50 mg twice/day orally was prescribed due to depression. After 1 week of nefazodone therapy the patient experienced headache, confusion, and "gray areas" in her vision, without abnormal ophthalmologic findings. Her serum creatinine was elevated to 2.2 mg/dl (baseline 1.5 mg/dl), and trough tacrolimus level was markedly elevated (> 30 ng/ml). Both tacrolimus and nefazodone are metabolized by the cytochrome P450 (CYP) 3A4 system. We suspect that nefazodone inhibits metabolism of tacrolimus. Coadministration of antidepressant agents such as nefazodone, or any other drug that inhibits the CYP3A4 isoenzyme subfamily, should be anticipated to interfere with tacrolimus metabolism. Monitoring blood concentrations of tacrolimus is vital, and appropriate dosage adjustments are required when the two drugs are administered concurrently to avoid serious interactions such as nephrotoxicity and neurotoxicity. PMID- 9855341 TI - Measurement of outcomes in adults receiving pharmaceutical care in a comprehensive asthma outpatient clinic. AB - We hypothesized that a pharmacist-provided comprehensive education program in conjunction with care provided by a pulmonologist would lead to improved economic, clinical, and humanistic outcomes in adults with asthma, compared with similar patients receiving care from a pulmonologist alone. The experimental group reported receiving more information about asthma self-management (p=0.001), were more likely to monitor peak flow readings (p=0.004), and had increased satisfaction with care, and perceived higher quality of care. Both groups had less lost productivity, fewer emergency department visits, fewer hospitalizations, and fewer physician visits, as well as improvement in symptoms scores within 45 days. Both groups improved in all functional status domains except the mental component score of the SF-12. Our results show a positive impact on outcomes in adults with asthma who received pharmaceutical care. PMID- 9855342 TI - Warfarin, short bowel syndrome, and intravenous lipids. PMID- 9855343 TI - Amiodarone: the re-emergence of a unique antiarrhythmic drug. Introduction. PMID- 9855344 TI - Oral amiodarone: historical overview and development. AB - OBJECTIVES: To review the historical development of amiodarone and the changing perceptions of the drug, and discuss its electrophysiologic, pharmacologic, and pharmacokinetic properties. METHODS: Review of relevant literature. RESULTS: In the 1970s and 1980s a plethora of new antiarrhythmic agents, including amiodarone, was introduced. Amiodarone is predominately a class III antiarrhythmic, but also possesses class I, II, and IV effects. By 1977 it was described as the ideal antiarrhythmic agent. However, clinicians underestimated potential difficulties caused by misunderstanding its variable absorption, slow initial response at nonloading dosages, and extended half-life. Elevated dosages also produced frequent adverse effects. Thus, early enthusiasm for the drug's efficacy was gradually replaced by a focus on its toxicity. The 1990s witnessed reacceptance of the agent as more logical initial regimens and lower maintenance dosages decreased adverse effects, and amiodarone emerged as one of the few drugs effective in suppressing and preventing arrhythmias that does not increase mortality. Remaining challenges include delineation of an optimal oral regimen, identification of markers useful in clinical monitoring, and elucidation of the relationship between dose-tissue concentration and response and dose-toxicity associations. CONCLUSION: Amiodarone is an increasingly valuable component of today's antiarrhythmic therapy. PMID- 9855345 TI - Clinical efficacy of amiodarone. AB - OBJECTIVES: To review the clinical efficacy and role of amiodarone in the management of supraventricular and ventricular arrhythmias and its effects on mortality. METHODS: Review of relevant studies and reports. RESULTS: Amiodarone exerts significant effects on atrial tissue. In most studies it was completely or partly effective in preventing recurrences of atrial fibrillation or flutter in up to 80% of patients. Amiodarone may be superior to class Ia agents for maintaining normal sinus rhythm. Large randomized trials indicate that it is a potent suppressor of ventricular arrhythmia and reduces arrhythmic death after myocardial infarction. In patients with cardiomyopathy, it suppresses asymptomatic arrhythmias and increases left ventricular ejection fraction. Meta analysis of relevant studies indicated that amiodarone reduces the risk of arrhythmic and sudden death by 29% in high-risk patients with recent myocardial infarction or congestive heart failure. This translates into an overall 13% reduction in total mortality. CONCLUSION: Because of its effectiveness against a broad range of arrhythmias, amiodarone is a valuable addition to the antiarrhythmic pharmacopeia. PMID- 9855346 TI - Optimal management of amiodarone therapy: efficacy and side effects. AB - OBJECTIVES: To review management and dosing guidelines for amiodarone therapy, and discuss the drug's adverse event profile. METHODS: Review of relevant studies and reports. RESULTS: Amiodarone is a highly effective antiarrhythmic drug, but is associated with adverse effects involving several organs. Amiodarone-induced arrhythmia is rare, with frequency of 0.3% in one study. Pulmonary toxicity is the most serious noncardiac side effect (2-17% of patients). Hypersensitivity pneumonitis can appear early in the course of therapy. Interstitial pneumonitis is a more common but insidious pulmonary reaction characterized by cough, low grade fever, and dyspnea that occurs after months or years of therapy. Clinically important hypothyroidism and hyperthyroidism occur in 2-10% of patients. Optic neuritis or neuropathy in which patients experience decreased or blurred vision may progress to permanent blindness. Abnormalities in liver function tests, especially elevated aminotransferase and alkaline phosphatase levels, are seen in 4-25% of patients. Neurologic side effects were reported in 20-40% of patients, at times associated with tremor, ataxia, peripheral neuropathy, malaise or fatigue, sleep disturbances, dizziness, and headaches. Several types of dermatologic reactions have been reported, including allergic rash, photosensitivity, and blue-gray skin discoloration. The best strategy for early detection of pulmonary toxicity is vigilant clinical follow-up with monitoring of cardiac status and liver and thyroid function, and prescription of the lowest effective dosage. After an initial loading dose, 200 mg/day in many patients maintains arrhythmia control and minimizes the frequency of side effects. CONCLUSION: Amiodarone is a safe and efficacious antiarrhythmic agent when lower dosages are given to patients who are closely monitored and subject to careful follow-up. PMID- 9855347 TI - Rationale, development, and clinical outcomes of a multidisciplinary amiodarone clinic. AB - OBJECTIVES: To review the rationale and development of a multidisciplinary amiodarone clinic, and document the clinical outcomes resulting from its implementation. METHODS: A clinic was established to provide an ambulatory setting in which patients receiving amiodarone could be followed according to published guidelines by a multidisciplinary team of cardiovascular health care specialists. Patients receiving amiodarone were referred to the clinic by their primary physicians. A data base containing each patient's medical history, current drug therapy, and baseline laboratory values was developed during the initial visit. Liver function tests, thyroid function tests, and chest radiographs were performed every 6 months, and pulmonary function tests were scheduled on an annual basis. Dosage adjustments were performed in select patients. RESULTS: Since November 1996, 60 patients have been referred to the amiodarone clinic. Mean length of follow-up before and after referral was 16.3+/ 25.5 and 9.2+/-5.5 months, respectively. Laboratory tests were performed according to accepted guidelines in 14 (23%) patients before referral compared with 54 (90%) patients after enrollment (p<0.001). Previously unrecognized adverse events were detected in 21 (35%) patients, including pulmonary fibrosis, QT prolongation, liver enzyme elevation, hypothyroidism, hyperthyroidism, and asthma exacerbation. Amiodarone was discontinued in six patients, four of whom had suspected pulmonary toxicity. The dose of amiodarone was adjusted in 29 (48.4%) patients. CONCLUSION: Many patients receiving amiodarone are not being followed according to published recommendations. Implementation of a specialized, multidisciplinary amiodarone clinic improves outcomes by monitoring for early detection of drug-related toxicities and by facilitating proper dosage modifications. PMID- 9855348 TI - Epithelioid hemangioendothelioma presenting in the skin: a clinicopathologic study of eight cases. AB - Epithelioid hemangioendothelioma arising in the skin is extremely rare, and the majority of documented cases have developed in association with an underlying bone tumor. We report eight patients with an age range of 29-84 years (mean 53), who presented with primary cutaneous tumors at a variety of sites including the palm, shin, neck, knee, nose, back, and penis with a duration of between 6 and 12 months. Histologically, all eight cases presented as circumscribed nodules with an overlying acanthotic epidermis, three showing striking acrosyringeal proliferation, reminiscent of eccrine syringofibroadenoma. The tumors were composed of an admixture of slightly pleomorphic spindle and epithelioid cells with abundant, sharply defined eosinophilic cytoplasm and vesicular nuclei containing single nucleoli. Mitoses were generally sparse. All tumors showed intracytoplasmic lumina and intraluminal erythrocytes were occasionally apparent. The tumor cells were embedded in a myxoid or hyaline matrix. In contrast to visceral lesions, a vascular origin was not evident in any of our cases. The tumor cells variably expressed CD31, CD34, factor VIII-Rag, and smooth-muscle actin but not pankeratin or epithelial membrane antigen. Follow-up ranged from 4 months to 3 years. None of the lesions has thus far recurred and there have been no metastases. PMID- 9855349 TI - Eosinophilic globules in spindle cell and epithelioid cell nevi: composition and possible origin. AB - The presence of eosinophilic globules has been described as a helpful sign for the histologic differentiation of Spitz's nevus from malignant melanoma. The origin of these bodies is not clear, and they have been likened to Civatte or colloid bodies of lichen planus. This would suggest that they might originate from degenerating keratinocytes or melanocytes or both. These eosinophilic globules and the colloid bodies of lichen planus have been reported to be similar in that they both stain positively for type IV collagen and laminin. These previous reports have failed to include, or have not emphasized, the staining for keratin that separates these two bodies. We stained 10 spindle cell and epithelial cell (S&E) nevi for S-100 protein, keratin, vimentin, type IV collagen, and laminin. In all 10 cases of S&E nevi, the eosinophilic globules showed a positive reaction for type IV collagen and laminin and a negative reaction for keratin, S100 protein, and vimentin, unlike the colloid bodies of lichen planus, which showed a negative reaction for type IV collagen and laminin and also a strong positive reaction for keratin. These results suggest that the eosinophilic globules of Spitz's nevi are basement membrane material, perhaps synthesized by either basal cells, melanocytes or both, and are not degenerated basal cells or melanocytes. PMID- 9855350 TI - Eosinophilic globules in spitz nevi: no evidence for apoptosis. AB - Eosinophilic globules are commonly found in Spitz nevi and can be of help in the differential diagnosis with melanoma. Despite their use as a diagnostic marker, there are conflicting concepts concerning the mechanism of the formation of these globules. Because pyknotic nuclei are often present in the cells surrounding the globules, many believe that apoptotic cell death of keratinocytes and melanocytes is an important mechanism in their formation. We studied nine Spitz nevi containing 21 eosinophilic globules for apoptotic cell death around and within Kamino bodies using the TUNEL method (TdT-mediated dUTP-x nick end-labeling method). In none of the cases were positive cells seen in the vicinity of the globules. Single apoptotic cells were found only sporadically in the melanocytes of the neoplasms (one to four per section) and were scattered throughout the entire lesion. The absence of apoptosis-specific labeled cells in the vicinity of eosinophilic globules and the rare occurrence of positive stained cells in other parts of the tumors makes apoptotic cell death unlikely to be relevant in the formation of eosinophilic globules. PMID- 9855351 TI - The melanocyte differentiation pathway in spitz nevi. AB - The nature of Spitz nevi is poorly understood, and their distinction from malignant melanoma can be difficult. Although there is general agreement on the diagnostic criteria, experts continue to have some differences, and controversial cases are not rare. A major obstacle to progress in this area is the lack of basic knowledge about melanocyte differentiation in Spitz nevi, as compared with ordinary nevi and malignant melanomas. Based on the hypothesis that normal melanocytes may have a differentiation pathway with discrete stages, it is suggested that the features of Spitz nevi may reflect homeostatic mechanisms governing maturation in the melanocyte differentiation pathway, whereas those of malignant melanomas may reflect carcinogen-induced aberrations. This perspective may be helpful in the continuing effort to develop optimal criteria for the differential diagnosis of Spitz nevi from malignant melanomas. PMID- 9855352 TI - Primary cutaneous adenoid cystic carcinoma with lymph node metastasis. AB - We report on a case of primary cutaneous adenoid cystic carcinoma with local recurrence and lymph node metastasis. The patient was a 52-year-old Japanese woman. The initial cutaneous lesion was a skin-colored, centrally elevated, thumb sized nodule with hair loss on the scalp in the right side of the parietal region. Three and half years after the initial combined treatment consisting of surgery and irradiation, a recurrent nodule on the left side of the parietal region of the scalp and a lymph node metastasis with perinodal tissue involvement in the left retroauricular area were discovered. In the literature on primary cutaneous adenoid cystic carcinoma, the average age in the 37 cases reported, including the present case, is 58.1 years, and 23 of the patients are women. Thirty-two percent of the carcinomas (12 cases) occur on the scalp and 16% (6 cases) on the breast. The average size of the tumors in the 24 documented cases is 3.2 cm. The local recurrence rate is 51% (19 cases). Distant metastasis to the lungs and pleura are recorded in three cases. Lymph node metastasis is recorded only in two cases including the present case. Multiple modalities including surgery, irradiation, and chemotherapy should be used for this locally aggressive and potentially metastasizing carcinoma. PMID- 9855353 TI - Pigmented squamous cell carcinoma of the skin: report of two cases and review of the literature. AB - Melanoma is the most common malignant tumor in which melanin synthesis occurs, although other nonmelanocytic tumors synthesize melanin or contain nonneoplastic melanocytes. We present two cases of infiltrating pigmented squamous cell carcinoma of the skin and review the clinical, morphologic, and ultrastructural features. Melanin was found in epithelial tumor cells as well as in macrophages and dendritic melanocytes. Interestingly, one of the neoplasms was associated with an adjacent melanocytic nevus and pigmented solar keratosis. Immunohistochemical analysis showed that neoplastic cells stained for keratin and melanin-filled dendritic cells were found to be S-100 protein and HMB45 positive. A careless examination of the immunohistochemical stains for S-100 protein and HMB45 could cause the misdiagnosis of melanoma, a neoplasm that has a more ominous outlook. PMID- 9855354 TI - Scrotal skin ulcer in a patient with a previous tonsillectomy because of natural killer cell lymphoma. AB - The CD56+ lymphomas are a recently characterized high-grade malignancy of putative natural killer cell origin. They are mostly localized to the nasal areas but show a propensity to spread to or recur in the skin. We describe a unique case of CD56+ natural killer lymphoma that recurred in scrotal skin in a patient 8 years after an initial limited resection. Although this case was unusual in showing a prolonged period of apparent remission, it illustrated a characteristic clinicopathologic behavior of this rare tumor. PMID- 9855355 TI - Anomalies associated with dye exclusion as a measure of axolemmal repair in invertebrate axons. AB - After axonal injury, dye exclusion is often used as a measure of the re establishment of a structural barrier. We now report that this use of dye exclusion is equivocal in two situations. (1) When a negatively-charged hydrophilic fluorescent dye (HFD) was placed in the physiological saline (PS) surrounding a crayfish medial giant axon (CMGA) before transection, this dye did not readily diffuse into the cut ends after transection whereas uncharged or neutralized dyes did do so. (2) When axoplasm flowed out of the cut ends of a transected squid giant axon (SGA), this outflow markedly slowed hydrophilic fluorescent dyes from diffusing into the cut ends. These anomalies suggest that dye exclusion by an injured axon does not always indicate that a structural barrier has formed. Therefore, dye assessments of axonal repair require control experiments that rule out anomalous exclusion due to dye interactions (biochemical and fluid dynamics) with components (axoplasm, axolemma, glial sheath, etc.) of the particular axon under study. PMID- 9855356 TI - Experimental pneumococcal meningitis in rabbits: the increase of matrix metalloproteinase-9 in cerebrospinal fluid correlates with leucocyte invasion. AB - Gelatinolytic activity of matrix metalloproteinases (MMPs), particularly MMP-9 and MMP-2, was studied by quantitative zymography in a rabbit model of bacterial meningitis during 24 h after inoculation with Streptococcus pneumoniae. In cerebrospinal fluid (CSF), MMP-2 was constitutively present and its level did not change during the experiment. In contrast, MMP-9, hardly detectable in CSF of healthy animals, increased dramatically. The increase of MMP-9 was correlated with both, an increase of CSF cell count and of total protein concentration. Intrathecal production of MMP-9 and MMP-2 was demonstrated by zymography of equal amounts of total protein from CSF and serum. Homogenates, prepared from various cortical regions of infected rabbits did not show increase of MMP activities. On the other hand, leucocytes isolated from CSF expressed high levels of MMP-9 suggesting a significant contribution of these cells to the elevation of MMP-9 activity in this body fluid. PMID- 9855357 TI - Insulin-induced hypoglycemia does not impair the surge of luteinizing hormone secretion in the proestrous rat. AB - To know whether insulin-induced hypoglycemia (IIH), which has been shown to inhibit pulsatile luteinizing hormone (LH) secretion, also affects the surge of LH secretion, adult female rats were injected with insulin (5 units/rat) or saline intravenously at 1300 or 1600 h on the day of proestrus and serum concentrations of LH and blood glucose were determined during the period from 1100 to 2100 h. The injection of insulin at neither 1300 nor 1600 h affected the surge of LH secretion, but it significantly decreased the blood glucose. Together with our recent hypothesis that the pulsatile and surge of LH secretion are controlled by separate gonadotropin-releasing hormone (GnRH) mechanisms, we conclude that the GnRH surge generator is less sensitive to IIH than the GnRH pulse generator. PMID- 9855358 TI - Different time courses of recovery after poisoning with botulinum neurotoxin serotypes A and E in humans. AB - Botulinum toxin serotypes A and E (BoNT/A and /E) cleave the carboxy-terminus of synaptosomal associated protein-25 (SNAP-25) removing nine and 26 residues, respectively. To investigate the effect of these lesions of the same target molecule, 11 volunteers were injected with 3 IU of BoNT/A in the extensor digitorum brevis (EDB) muscle of one foot and with 3 IU of BoNT/E in the contralateral one. In addition, seven volunteers were similarly injected with mixtures of BoNT/A + BoNT/E. Compound muscular action potential (CMAP) was measured at different time intervals and the percentage variation of CMAP (%CMAP) was calculated. Unexpectedly, a much faster recovery of %CMAP after BoNT/E injections was observed. Double poisoned EBD muscles recovered similarly to BoNT/E. So, a larger deletion of the SNAP-25 molecule caused by BoNT/E leads to a faster functional recovery. PMID- 9855359 TI - Activation of dihydropyridine sensitive Ca2+ channels in rat hippocampal neurons in culture by parathyroid hormone. AB - We examined the effects of parathyroid hormone (PTH) on rat hippocampal neurons in culture to determine whether it caused a similar intracellular calcium concentration ([Ca2+]i) increase in these cells to that seen with renal epithelial cells and found that PTH induced the effect in about 30% of the neurons. The effects appeared gradually during continuous administration of full length PTH(1-84) or its active fragment, PTH(1-34), but not of an inactive fragment, PTH(39-84). However, the active fragment of the PTH-related peptide (PTHrP(1-34)) had little effect on [Ca2+]i during 60 min of administration. The PTH effect was inhibited by nifedipine, an L-type Ca2+ channel antagonist, and facilitated by S-(-)-BAY K 8644, an L-type Ca2+ channel agonist. Our findings suggest that PTH is one of the causal factors for the age-related increase in the density of voltage gated Ca2+ channels in hippocampal neurons. PMID- 9855360 TI - Molecular modulation of recombinant rat alpha1beta2gamma2 GABA(A) receptor channels by diazepam. AB - Recombinant gamma-aminobutyric acid (GABA(A)) receptor channels containing alpha1beta2gamma2-subunits were transiently expressed in HEK293 cells. Modulation by diazepam (DZ) was investigated using the patch-clamp technique with a device for ultra-fast solution exchange. GABA activated Cl(-)-currents were potentiated when DZ > 0.1 microM was added to non-saturating concentrations of GABA (< 0.1 mM GABA). Maximal potentiation of the peak current amplitude by a factor of 2.5 was observed when 1 microM DZ was added to the test-solution. Deactivation of GABA activated currents after the end of GABA pulses was best fitted with two time constants. After application of DZ + GABA, increase of time constants of deactivation was measured. It was independent on GABA concentration. We conclude that prolongation of deactivation after application of GABA + DZ may be an important mechanism of the modulatory action of DZ at GABA(A) receptor channels. PMID- 9855361 TI - Glucose transporter GLUT1 localization in human foetus telencephalon. AB - The endothelial cells of the mature cerebral microvessels, provided with barrier devices (blood-brain barrier, BBB), selectively express the glucose transporter isoform 1 (GLUT1). Presence and localization of the GLUT1 were studied by immunogold silver staining (IGSS) labelling on ultrathin sections of foetal human telencephalon tissue embedded in Lowicryl HM20 according to the progressive lowering of temperature (PLT) method. In the microvascular endothelial cells of the human telencephalon GLUT1 molecules are detected at the 12th gestational week and their expression is increased at the 18th week. In both ages, the transporter is mainly localized on the ablumenal and lateral endothelial cell membranes, and at 18 weeks a greater number of GLUT1 antigenic sites are also seen at the lumenal membrane. Our findings demonstrate both the expression and subcellular localization of GLUT1 be developmentally regulated and suggest an early functioning of the BBB-GLUT1 transporter in the developing human brain. PMID- 9855362 TI - Immunohistochemical localization of nicotinic beta2 and alpha4 receptor subunits in normal human brain and individuals with Lewy body and Alzheimer's disease: preliminary observations. AB - Optimum immunohistochemical methods were established to immuno-localize nicotinic acetylcholine receptor alpha4 and beta2 subunits in temporal cortex and substantia nigra of normal aged and diseased human brain. In normal aged brain, fibers were immunoreactive for both the alpha4 and beta2 subunits of the nicotinic receptor in the temporal cortex and the substantia nigra. In the cortex of normal aged brain, rare neurofibrillary tangles occurring could be identified with either anti-alpha4 or anti-beta2 antibodies, but existing senile plaques were demonstrable with neither. In Alzheimer's disease temporal cortex, there were diminished numbers of nicotinic receptor subunit immunoreactive fibers, and there were appreciable numbers of neuropil threads, neurofibrillary tangles and senile plaques immunoreactive with both the alpha4 and beta2 antibodies. PMID- 9855363 TI - Density of motility-related charge in the outer hair cell of the guinea pig is inversely related to best frequency. AB - Whole cell voltage clamp and freeze fracture were used to study the electrophysiological and ultrastructural correlates of the outer hair cell (OHC) lateral membrane molecular motors. We find that specific voltage-dependent capacitance, which derives from motility-related charge movement, increases as cell length decreases. This increasing non-linear charge density predicts a corresponding increase in sensor-motor density. However, while OHC lateral membrane particle density increases, a quantitative correspondence is absent. Thus, the presumed equivalence of particle and motor is questionable. The data more importantly indicate that whereas the voltage driving OHC motility, i.e. the receptor potential, may decrease with frequency due to the OHC's low-pass membrane filter, the electrical energy (Q x V) supplied to the lateral membrane will tend to remain stable. This conservation of energy delivery is likely crucial for the function of the cochlear amplifier at high frequencies. PMID- 9855364 TI - Localization of alpha-tocopherol transfer protein in rat brain. AB - Vitamin E (alpha-tocopherol) is a fat-soluble antioxidant that is transported by plasma lipoproteins in the body. Alpha-tocopherol transfer protein (alpha-TTP), which was identified as a product of the causative gene for familial isolated vitamin E (FIVE) deficiency, is a cytosolic liver protein which plays an important role in the efficient circulation of plasma vitamin E in the body. In the present study, we detected the message for alpha-TTP at low levels in some rat tissues including brain, spleen, lung and kidney. In the brain, the alpha-TTP transcript was detected predominantly in the cerebellar cortex, as revealed by in situ hybridization histochemistry. In the cerebellar cortex, clusters of the hybridization signal were aligned with the Purkinje cell layer, although the signal was not detected in the Purkinje cells, but in the small cells around the Purkinje cells. This distribution pattern strongly suggests that the message for alpha-TTP is expressed in the Bergmann glial cells. Combined with the previous observation that there was severe Purkinje cell loss in patients with FIVE deficiency, the present data suggest that vitamin E is supplied to the Purkinje cells from the surrounding Bergmann glial cells with the help of alpha-TTP. PMID- 9855365 TI - The correction of alpha7 nicotinic acetylcholine receptor concentration-response relationships in Xenopus oocytes. AB - The rapid desensitization of alpha7 nicotinic acetylcholine receptors (nAChR) has presented a serious problem for the characterization of this receptor subtype, potentially confounding the interpretation of concentration-response relationships. However, the consistency of cell geometry and solution flow in oocyte recordings permits estimations of instantaneous concentrations to be made in this system. Results interpreted with predicted instantaneous concentrations suggest that estimates of EC50 derived from conventional analysis may overestimate the actual EC50 values by a factor of 10 and underestimate Hill slopes by a factor of 2-3. If the limiting desensitization process of alpha7 receptors is driven by the agonist concentration itself rather than by time dependent processes, then similar dependencies may exist between the response and instantaneous agonist concentration in other systems. PMID- 9855366 TI - Interleukin-10 stimulation of corticotrophin releasing factor median eminence in rats: evidence for dependence upon nitric oxide production. AB - Following treatment with interleukin-10 (IL-10), basal corticotrophin releasing factor (CRF) levels from rat hypothalamic median eminence (ME) were found to be increased. Our data show: (1) the specificity of stimulation of CRF through the use of recombinant IL-10 and its blockage by monoclonal anti-IL-10 antibody; (2) the requirement of NO in this process through the use of N(omega)-nitro-L arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor; (3) the blockage of IL-10 stimulated NO production by anti-IL-10; and, (4) the presence of IL-10 transcripts in hypothalamic poly A+ mRNA. These results provide the first evidence of IL-10 acting in the ME to influence CRF levels and further support our earlier findings of a potential for IL-10 in the hypothalamic pituitary axis. PMID- 9855367 TI - The effects of pulsing magnetic fields on pineal melatonin synthesis in a teleost fish (brook trout, Salvelinus fontinalis). AB - Based on findings in various mammalian species, where exposure to electromagnetic fields decreased the nocturnal synthesis of the pineal secretory product melatonin, we investigated the effects of magnetic field (MF) exposure in a teleost fish, the brook trout (Salvelinus fontinalis). Fields were generated by Helmholtz coils (maximum flux density 40 microT, frequency 1 Hz, 200 ms on, 800 ms off). Melatonin concentrations were estimated by a specific radioimmunoassay. MF exposure significantly increased night-time pineal (P < 0.001) and serum (P < 0.01) melatonin levels, as compared with the controls. It is suggested that either the pineal glands are directly affected, i.e. by an increased Ca2+-influx into pineal photoreceptors, or that the responses are indirect since induced currents, caused by the rapid rise and decay of the generated MF, may have disturbed the sensory system for electric fields. PMID- 9855368 TI - No increase in cerebrospinal fluid tau protein levels in patients with vascular dementia. AB - Tau protein levels in cerebrospinal fluid (CSF-tau) were determined in 29 patients with old cerebrovascular disease (CVD, 21 demented and eight non demented), 69 patients with Alzheimer's disease (AD) and 17 age-matched normal controls. The CSF-tau level in the vascular dementia (VD) group (24.0 +/- 17.0 pg/ml) was significantly lower (P < 0.0001) than that in the AD group (90.0 +/- 45.3 pg/ml), but not significantly different from that in the non-demented patients with CVD (18.1 +/- 10.2 pg/ml) or controls (20.3 +/- 13.0 pg/ml). Among the VD patients, 1/21 exceeded a cut-off value (mean +/- 2 SD of controls), whereas 8/69 of the AD patients had CSF-tau levels below this value. These findings suggest that VD constitutes a group of dementias that can be separated from AD by normal CSF-tau levels. CSF-tau determinations in combination with other clinical findings may provide another diagnostic aid in the differential diagnosis between VD and AD. PMID- 9855369 TI - Expression of the nicotinic acetylcholine receptor alpha4 subunit mRNA in the rat cerebellar cortex. AB - In order to compare the previous immunohistochemical and immunocytochemical data on the distribution of nicotinic acetylcholine receptor alpha4 subunit-like immunoreactivity with the expression of alpha4 mRNA in the rat cerebellar cortex, the present study determined the cellular distribution of alpha4 mRNA in the rat cerebellar cortex. Northern blot analysis revealed two alpha4 mRNA bands in the rat cerebellum and three in the cerebral cortex, hippocampus, hypothalamus and striatum. The total level of these transcripts was lower in the cerebellum than in the other four regions. The expression of alpha4 mRNA was high in Purkinje cells and granular cells, whereas low expression was detected in the molecular layer. These results suggest that the expression of alpha4 mRNA is closely related to the alpha4-like immunoreactivity in the molecular and Purkinje cell layers. In the granular layer, alpha4 mRNA was very highly and broadly expressed in comparison with the alpha4-like immunoreactivity. PMID- 9855370 TI - Endothelial derived vasoactive factors and leukocyte derived inflammatory mediators in subjects with asymptomatic atherosclerosis. AB - To clarify relationships between the (endothelial vasodilatory and vasoconstrictive function) and leukocyte inflammatory mediators in subjects with asymptomatic atherosclerosis, we measured (intraplatelet cyclic 3',5'-guanosine monophosphate [cGMP] and cyclic 3',5'-adenosine monophosphate [cAMP]), plasma endothelin (ET-1), and plasma neopterin in 197 subjects with asymptomatic atherosclerosis (median age 63 years, range 49-69 years). We measured neutrophil protease 4 (NP4), tumor necrosis factor (TNFmu), soluble tumor necrosis factor receptor-1 (sTNFR-1), and neutrophil gelatinase associated lipocalin (NGAL) in 152 of the 197 subjects. Intraplatelet cGMP correlated inversely with plasma ET-1 (r=-0.22; p=0.01), which confirms earlier in vitro data of the inhibitory effect of ET-1 on NO production and/or the cGMP mediated inhibitory effect of NO on ET-1 production. Plasma neopterin as well as NP4 correlated directly with intraplatelet cGMP (r=0.24; p<0.01 and r=0.33; p<0.001, respectively). Intraplatelet cAMP correlated directly with plasma TNFmu (r=0.17; p<0.05) and sTNFR-1 (r=0.20; p<0.05). The relationship between leukocyte derived inflammatory mediators and intraplatelet cyclic nucleotides suggest an antiaggregating effect of leukocytes upon platelets, which may constitute a negative feedback mechanism that inhibits platelet activation during the atherosclerotic inflammatory process. PMID- 9855371 TI - Left ventricular size and function in patients with noninsulin-dependent diabetes and postinfarction total or subtotal coronary occlusions. AB - The aim of this study was to establish the effects of postinfarction total or subtotal coronary occlusion on left ventricular remodeling in patients with noninsulin-dependent diabetes (NIDD) compared with the effects in postinfarct nondiabetic patients. The authors selected 100 patients submitted to coronary angiography between 1 and 5 weeks after acute myocardial infarction (T0: 20.5+/ 15.4 days) and classified into three groups: G1: NIDD with coronary occlusion/subocclusion (n=24), G2: controls with coronary occlusion/subocclusion (n=43), G3: controls without coronary occlusion/subocclusion (n=33). At time zero (T0) the following parameters were evaluated: end-systolic and end-diastolic volume indexes (ESVi, EDVi), ejection fraction (EF), echocardiographic wall motion score index (WMI), presence of left ventricular aneurysm, and triple vessel coronary disease. The frequencies of major cardiovascular events were recorded during follow-up. Significantly greater ESVi and EDVi were noted in G2 compared with G3 (P<0.0001), while no significant differences were observed between NIDD patients and controls. Although left ventricular global and segmental dysfunctions were increased in diabetics, controls with coronary occlusion/subocclusion presented more pronounced EF reduction (P<0.0001 G2 vs G3) and higher elevation in WMI (P<0.005 G2 vs G3). Cardiac events during follow-up were elevated in G1 and G2, particularly as regards the occurrence of congestive heart failure. The authors conclude that NIDD seems to influence in a positive way left ventricular remodeling associated with postinfarct total or subtotal coronary occlusion. PMID- 9855372 TI - An improved method for coronary sinus blood flow monitoring by intravascular Doppler technique. AB - In order to obtain accurate measurements of coronary sinus blood flow (CSBF), a new catheter (7 French) with a radiopaque, flexible, and basket-shaped tip was developed for guiding a standard 3 Fr Doppler catheter in the coronary sinus (CS) in man. The radiopaque "basket" tip of the catheter allows the operator to stabilize the position of the Doppler transducer in the center of the CS and to accurately measure the CS internal diameter radiologically. CSBF was calculated as the product of CS cross-sectional area by mean CSBF velocity. Doppler-derived CSBF values at rest and during handgrip were compared with those obtained by the local thermodilution technique in 16 patients undergoing diagnostic coronary angiography. During handgrip, mean CSBF increased from 154+/-23 (rest) to 299+/ 34 mL/min by the Doppler method and from 148+/-22 to 288+/-32 mL/min by the thermodilution technique. A good correlation (r = 0.86) between the CSBF values with the two techniques was observed. The authors conclude that the intravascular Doppler technique associated with the use of the basket guide catheter provides an accurate and simple tool for monitoring CSBF in patients. PMID- 9855373 TI - The effect of indirect calorimetry measurement on claudication pain in patients with peripheral arterial occlusive disease. AB - The effect of indirect calorimetry measurement on the duration to onset and maximal claudication pain and hemodynamic measures was evaluated. Fifteen male patients with peripheral arterial occlusive disease (PAOD) performed two graded exercise tests (GXTs) in random order, one with and the other without the use of indirect calorimetry measurement. During the tests the time to onset and maximal claudication pain was recorded. Before and after the tests, ankle/brachial systolic pressure index (ABI) was determined. The time to onset of claudication pain was shorter for the test with indirect calorimetry (4:49+/-3:50 vs 5:44+/ 4:39 min:sec, p < 0.005). Time to maximal claudication pain was not significantly different between the two conditions (8:58+/-3:01 min:sec with indirect calorimetry, 9:39+/-3:36 min:sec, without). There were no significant differences in ABI between the two testing sessions at any time point. Results of this study suggest that indirect calorimetry measurement can be utilized without compromising maximal claudication pain time or hemodynamic measurements but that time to onset of claudication pain is hastened with indirect calorimetry measurement in patients with PAOD. PMID- 9855374 TI - Staged reperfusion preserves the coronary flow reserve, especially in the regions not severely damaged by ischemic injury in the canine heart. AB - The objective of this study was to determine the effects of staged reperfusion on the progressive reduction in coronary blood flow (CBF) and coronary flow reserve during reperfusion and on the infarct size in the canine heart. Fifteen dogs underwent 90 min of left circumflex coronary artery occlusion and 3 hr of reperfusion. In the abrupt reperfusion group, the occluder was released completely at the initiation of reperfusion. In the staged reperfusion group, CBF was maintained at 20% of preocclusion values for 10 min after initiation of reperfusion, then gradually released, and completely released 20 min after initiation of reperfusion. There was no significant difference in CBF between the staged (n = seven) and abrupt (n = eight) groups after 3 hr of reperfusion. The repayment of flow debt in the staged reperfusion group was significantly greater than in the abrupt reperfusion group after 3 hr of reperfusion (260+/-120% vs 100+/-60%, staged vs abrupt at 3 hr, p < 0.03). The ratio of peak reactive hyperemic flow to resting flow in the staged reperfusion group was significantly greater than in the abrupt reperfusion group throughout the reperfusion phase (4.4+/-1.0 vs 2.6+/-0.6 at 3 hr, p < 0.001), and had returned to the preocclusion values after 3 hr of reperfusion. This preservation of the coronary flow reserve in the staged reperfusion group was observed in the epicardium (4.1+/-0.6 vs 2.8+/-0.7, staged vs abrupt at 3 hr, p < 0.01), but not in the endocardium or midmyocardium. Infarct size did not differ significantly between the two groups. Staged reperfusion in this study did not appear to attenuate the reduction of CBF, or to reduce infarct size, however preserved the coronary flow reserve, especially in the regions not severely damaged by ischemic injury. PMID- 9855375 TI - Role of human immunodeficiency virus in primary pulmonary hypertension--case reports. AB - Previous cases of pulmonary hypertension (PH) in human immunodeficiency virus (HIV) infection have been reported in the literature. The role of HIV in PH is still debatable. The purpose of this report was to analyze whether HIV plays a direct or indirect role in PH pathogenesis. Between February and November 1997, 56 HIV-infected patients with cardiac symptoms and signs were studied by serial color Doppler echocardiography. In four patients (7.1%), PH not related to other well-known associated conditions, was disclosed. In spite of a low serum HIV RNA viral load and a high-efficacy antiretroviral therapy, including a protease inhibitor in two patients, PH developed and worsened. It could be hypothesized that in some patients with an individual immunogenetic predisposition, a high secretion of cytokines and endothelin-1 stimulated by an unidentified pathogen different from HIV could lead to PH. Antiretroviral therapy seems not to prevent or reduce right ventricle pressure gradient in PH. PMID- 9855376 TI - Catheter fracture and embolization from totally implanted venous access ports- case reports. AB - Totally implanted venous access ports are excellent devices for delivering chemotherapeutic agents and prolonged intravenous infusions in patients with cancer. Catheter fracture and embolization are rare and potentially serious complications of these widely used devices. Retrieval of the embolized fragment is generally indicated but may not be possible. The authors report three cases of catheter embolization in their center over a period of 9 years. Catheter "pinch off," fracture, embolization, and retrieval are discussed. PMID- 9855377 TI - Left gastric artery arising from the superior mesenteric artery--case reports. AB - Two case reports of a left gastric artery replaced to the SMA are presented. Normal left gastric anatomy and development are described, and the importance of such anomalies is discussed. PMID- 9855378 TI - Ethics and managed care: reconstructing a system and refashioning a society. PMID- 9855379 TI - Managed care and end-of-life decisions: learning to live ungagged. PMID- 9855380 TI - Case of the month: not-so-obvious uses of the autopsy. Autopsy Committee of the College of American Pathologists. PMID- 9855381 TI - Lifestyle and 15-year survival free of heart attack, stroke, and diabetes in middle-aged British men. AB - BACKGROUND: To examine the relationship between modifiable lifestyle factors (smoking, physical activity, alcohol intake, and body mass index [BMI]) and the likelihood of 15-year survival free of major cardiovascular end points and diabetes in middle-aged men. METHODS: A prospective study of 7142 men aged 40 to 59 years at screening with no history of coronary heart disease, diabetes, and stroke drawn from 1 general practice in each of 24 British towns and followed up for 15 years. MAIN OUTCOME MEASURES: Death from any cause and a combined end point, including survival free of heart attacks or stroke or the development of diabetes over a follow-up of 15 years for each man. RESULTS: During the 15-year follow-up, there were 1064 deaths from all causes, 770 major heart attacks (fatal and nonfatal), 247 stroke events (fatal and nonfatal), and 252 cases of diabetes among the 7142 men. After adjustment for age and each of the other modifiable lifestyle factors, the risk of the combined end point (death or having a heart attack, stroke, or diabetes) went up significantly with increasing smoking levels and from BMI levels of 26 kg/m2 or higher, and decreased significantly with increasing levels of physical activity up to levels of moderate activity with no further benefit thereafter (heavy smoking vs never: relative risk [RR] [odds], 2.50; 95% confidence interval [CI], 2.12-2.94; BMI > or = 30 vs 20-21.9 kg/m2: RR, 2.11; 95% CI, 1.71-2.62; moderate vs inactive: RR, 0.60; 95% CI, 0.50-0.72). Light drinking (vs occasional) showed a relatively small but significant reduction in risk (RR, 0.84; 95% CI, 0.74-0.96). Using Cox predictive survival models, the estimated probability of surviving 15 years free of cardiovascular events and diabetes in a man aged 50 years ranged from 89% in a moderately active man at BMI levels of 20 to 24.0 kg/m2 who had never smoked to 42% in an inactive smoker with BMI level of 30 kg/m2 or higher. CONCLUSIONS: Modifiable lifestyles (smoking, physical activity, and BMI) in middle-aged men play an important role in long-term survival free of cardiovascular disease and diabetes. These findings should provide encouragement for public health promotion directed toward middle aged men. PMID- 9855382 TI - Digoxin toxicity: an evaluation in current clinical practice. AB - BACKGROUND: Serum digoxin concentrations (SDCs) are frequently sampled before completion of drug distribution. If elevated, these concentrations may be misinterpreted, potentially leading to a misdiagnosis of digoxin toxicity. OBJECTIVES: To determine the frequency of elevated SDCs (>2.6 nmol/L [>2.0 ng/mL]) obtained at appropriate postdosing intervals and to evaluate the frequency of clinically defined digoxin toxicity in patients with elevated SDCs. METHODS: The medical records of adult patients with SDCs assayed at 5 general hospitals in North Carolina during a 3-month period (May 1 through July 31, 1996) were prospectively evaluated. Data on SDC, inpatient or outpatient status, and medical or surgical service were collected for all patients. Data on patient demographics, serum chemistry values, indication for digoxin treatment, clinical evidence of digoxin toxicity, and timing of the blood sample relative to administration of the last dose of digoxin were collected for patients with SDCs higher than 2.6 nmol/L (>2.0 ng/mL). RESULTS: Of 3434 SDCs assayed in 2009 patients, 320 (9.3%) were higher than 2.6 nmol/L (>2.0 ng/mL). Fifty-one (15.9%) of the 320 SDCs were drawn at 6 hours or less following a digoxin dose. Sampling time relative to the digoxin dose could not be determined in 70 (21.9%) of the 320 elevated SDCs, leaving 199 (62.2%) of 320 SDCs in 138 patients evaluable for digoxin toxicity. Eighty-three of the 138 patients had clinical evidence of digoxin toxicity for an overall incidence of 4.1%. CONCLUSIONS: Digoxin toxicity occurs less frequently than historically reported. Continued emphasis needs to be placed on obtaining appropriately timed SDCs. PMID- 9855383 TI - Respiratory tract viral infections in inner-city asthmatic adults. AB - BACKGROUND: Respiratory tract viral infections (RTVIs) have been identified frequently in association with asthma exacerbations in children, but few studies have shown similar rates of viral infections in adults with asthma. Further studies using newer diagnostic techniques to evaluate the frequency of RTVIs in adults with acute exacerbations of asthma need to be performed. METHODS: Twenty nine asthmatic adults were recruited from the pulmonary clinic of an urban county hospital and were followed up in a longitudinal cohort study for signs and symptoms of asthma and RTVI. One hundred twenty-two asthmatic adults presenting to the emergency department (ED) of the same hospital with acute symptoms of asthma underwent evaluation for RTVI in a cross-sectional prevalence study. In both studies, respiratory secretions and paired serum samples were collected from subjects with acute wheezing episodes and evaluated using virus culture, serologic testing, and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In the longitudinal cohort study, 138 respiratory illnesses, of which 87 were asthma exacerbations, were evaluated; 41% of all illnesses and 44% of asthma exacerbations were associated with an RTVI. In the ED study, 148 asthma exacerbations were evaluated; 55% were associated with an RTVI. An RTVI was identified in 21 (50%) of 42 of the subjects hospitalized in the ED study. Picornaviruses (rhinoviruses), coronaviruses, and influenza viruses were the most commonly identified causes of RTVI. Forty-six (60%) of the 77 picornavirus infections and 22 (71%) of the 31 coronavirus infections were identified only using RT-PCR. CONCLUSIONS: Asthmatic exacerbations in adults are frequently associated with an RTVI. Identification of such infections often requires newer diagnostic methods, such as virus-specific RT-PCR. The high frequency of RTVIs identified in association with asthmatic exacerbations in adults from the inner city suggests that strategies for the prevention of RTVI should be targeted toward this population. PMID- 9855384 TI - Mortality of the institutionalized old-old hospitalized with congestive heart failure. AB - BACKGROUND: Congestive heart failure is a major cause of mortality and morbidity in the elderly but the disease impact on the oldest and sickest population has not been defined. OBJECTIVES: To review the mortality and hospital readmission rate of institutionalized elderly persons with congestive heart failure and to examine the relation of baseline characteristics to subsequent clinical outcomes. METHODS: This was a retrospective analysis based on chart review of 231 residents of the Philadelphia (Pa) Geriatric Center (63 congregate housing tenants and 168 nursing home residents) 80 years and older, hospitalized with congestive heart failure from 1989 to 1995. Patients' demographic data and clinical, electrocardiographic, and echocardiographic findings were obtained from their initial (index) hospitalization records. Subsequent outcomes were obtained from their outpatient (nursing home or office) records. RESULTS: Thirteen percent died during the index hospitalization but the total mortality during the follow-up period was 87%. One hundred forty-six patients (63%) died in the first year with a mean +/- SD survival of 4+/-4 months and a readmission rate of 3.9 per patient year. Eighty-five patients survived the first year with a readmission rate of 1.2 per patient-year and 54 patients subsequently died, with a mean +/- SD survival of 28+/-12 months. The first-year decedents and survivors were comparable in sex, age, medical history, and electrocardiographic findings. However, patients who died in the first year, compared with survivors, were more likely to be nursing home residents (81% vs 59%), have New York Heart Association class IV heart failure (54% vs 32%), have impaired left ventricular function by echocardiogram (53% vs 32%), and have renal insufficiency (32% vs 11%). CONCLUSIONS: Very elderly persons with congestive heart failure had a guarded long-term prognosis. Nursing home residency, class IV heart failure, impaired left ventricular function, and renal insufficiency were associated with higher risk for early death and repetitive hospitalizations. PMID- 9855385 TI - Identifying patients with depression in the primary care setting: a more efficient method. AB - OBJECTIVE: To determine if there is a core subset of depressive symptoms that could be used to efficiently diagnose depression after administering the 2-item PRIME-MD a screening questionnaire for depression. METHODS: One thousand patients selected randomly and by convenience from 4 primary care clinics were assessed by PRIME-MD and completed a questionnaire measuring the following validation variables: functional status and well-being, disability days, somatic symptoms, depression severity, suicidal thoughts, health care utilization, and the physician-patient relationship. RESULTS: Four symptoms (sleep disturbance, anhedonia, low self-esteem, and decreased appetite) accounted for virtually all the depression symptom-related variance in functional status and well-being, with 8.3% of patients having 2 of these symptoms and 8.2% having 3 or 4 of these symptoms. There was excellent agreement between diagnosis based on core symptoms and major depression (K= 0.77; overall accuracy rate, 94%). There were significant differences (P<.001) among patients with negative depression screen, 0 to 1, 2, and 3 to 4 core symptoms with scores on each of the validation variables getting progressively worse in these 4 groups. A cutoff point of 2 core symptoms identified all but 3 patients with major depression and an additional 5% of the entire sample without major depression who were significantly (P<.05) worse than patients without depression on each of the validation variables. CONCLUSION: A strategy that includes the use of a 2-item depression screener followed by the evaluation of 4 core depressive symptoms is an efficient and effective way of identifying and classifying primary care patients with depression in need of clinical attention. PMID- 9855386 TI - Diet and exercise in the treatment of obesity: effects of 3 interventions on insulin resistance. AB - BACKGROUND: In short-term studies, diet and exercise both improve insulin sensitivity. OBJECTIVE: To determine the effects of a 48-week supervised diet and exercise program on weight and insulin sensitivity after initial weight loss and weight maintenance, and then subsequent weight regain over 96 weeks. METHODS: Forty-five obese women were randomly assigned to 1 of 3 treatment groups: (1) diet alone, (2) diet and aerobic training, and (3) diet and strength training. All subjects received the same 48-week group behavior modification program and diet (approximately 3879 kJ/d [approximately 925 kcal/d] for the first 16 weeks; approximately 6276 kJ/d [approximately 1500 kca/d] thereafter). Exercising subjects were provided 3 supervised exercise sessions per week for the first 28 weeks and 2 sessions weekly until week 48. During weeks 48 to 96, subjects were unsupervised. Oral glucose tolerance tests were performed at baseline and weeks 16, 24, 44, and 96. RESULTS: Subjects across the 3 conditions achieved a mean weight loss of 13.8 kg by week 16, which was associated with decreased insulin levels (61.8% of baseline) There were no significant differences among groups in changes in body mass index, which is a measure of weight in kilograms divided by the square of the height in meters, weight, glucose tolerance, or insulin levels at weeks 16, 24, and 44. No additional beneficial effect of aerobic or strength exercise on insulin resistance, as reflected by serum insulin levels before and after a glucose load, was demonstrated. The 22 subjects who were studied at week 96 maintained a loss of approximately 10% of initial weight. Insulin levels, however, had returned to pretreatment levels. CONCLUSIONS: This study confirms the beneficial effect of weight reduction on hyperinsulinemia in obese individuals. Participation in supervised exercise did not result in additional improvement in weight loss or insulin sensitivity. We also observed a marked increase in insulin levels with only partial weight regain. Determining the amount of sustained weight loss necessary for continued improvement in insulin sensitivity will require further study. PMID- 9855387 TI - Effect of intensive glycemic control on fibrinogen, lipids, and lipoproteins: Veterans Affairs Cooperative Study in Type II Diabetes Mellitus. AB - BACKGROUND: The Veterans Affairs Cooperative Study in Type II Diabetes Mellitus prospectively studied insulin-treated patients with type 2 (non-insulin dependent) diabetes mellitus, achieving 2.1% glycosylated hemoglobin separation between intensive- and standard-treatment arms (P<.001) for 2 years. OBJECTIVE: To assess the effect of intensive therapy on serum fibrinogen and lipid levels, compared with standard treatment. METHODS: One hundred fifty-three male subjects with type 2 diabetes mellitus and who required insulin treatment were recruited from 5 Veterans Affairs medical centers. The subjects were divided into intensive and standard-treatment arms for a randomized prospective study. Dyslipidemia was managed identically in both arms (diet, drugs). Fibrinogen levels and lipid fractions were measured in the full cohort. Lipid fractions are separately reported in patients not treated with hypolipidemic agents. RESULTS: There were no baseline differences between arms. Fibrinogen levels rose in the intensive treatment arm at 1 year (from 3.34+/-0.12 to 3.75+/-0.15 g/L; P<.001) but returned to baseline at 2 years (3.47+/-0.12 g/L). There was no change in the standard-treatment arm. Triglyceride levels decreased in the intensive-treatment arm from 2.25+/-0.27 to 1.54+/-0.14 mmol/L (199+/-24 to 136+/-12 mg/ dL) at 1 year (P = .004) and to 1.74+/-0.18 mmol/L (154+/-16 mg/dL) at 2 years (P = .03); there was no change in the standard-treatment arm. Cholesterol levels decreased in the intensive-treatment arm at 1 year from 5.4+/-0.21 to 4.99+/-0.13 mmol/L (207+/-8 to 193+/-5 mg/dL) (P = .02); there was no change in the standard treatment arm. Levels of low- and high-density lipoprotein cholesterol decreased in the standard-treatment arm only by 2 years, from 3.44+/-0.13 to 3.16+/-0.10 mmol/L (133+/-5 to 122+/-4 mg/ dL) (P =.02) and from 1.10+/-0.03 to 1.00+/-0.03 mmol/L (42+/-1 to 38+/-1 mg/dL) (P<.001) for low-density and high-density lipoprotein cholesterol, respectively. Levels of apolipoprotein B decreased in both treatment arms (P<.001), and apolipoprotein A1 levels decreased in the standard-treatment arm (P<.01). Lipoprotein (a) levels did not change in either treatment arm. Lipid results were essentially identical whether examined in the full cohort or excluding those patients receiving hypolipidemic agents. CONCLUSIONS: Intensive insulin therapy led to a potentially beneficial reduction in serum triglyceride levels and preservation of high-density lipoprotein cholesterol and apolipoprotein A1 levels. However, it caused transient elevation in plasma fibrinogen levels, a possible thrombogenic effect. PMID- 9855388 TI - Barriers to completion of health care proxies: an examination of ethnic differences. AB - BACKGROUND: Advance directives have not been uniformly used by different segments of the US population and studies have consistently shown a lower prevalence of advance directives among African Americans and Hispanics compared with non Hispanic whites. OBJECTIVE: To examine barriers to completion of health care proxies for different ethnic groups. METHODS: One hundred ninety-seven subjects aged 65 years or older self-identified as African American (n = 65), Hispanic (n = 65), or non-Hispanic white (n = 67) attending a geriatrics and internal medicine outpatient clinic of a large New York City teaching hospital were administered a questionnaire. Questionnaires were developed to examine potential barriers to completion of health care proxies. Barriers were drawn from the literature and from focus groups. RESULTS: Significant predictors of proxy completion using logistic regression analysis included knowledge of health care proxies, availability of a health care agent, exposure to mechanical ventilation, age, and self-reported health status as fair to poor. Subjects who believed that a health care agent was irrelevant in the setting of involved family were significantly less likely to have completed a health care proxy. Although there were significant differences in the baseline completion rates of health care proxies for the 3 ethnic groups, ethnicity did not predict prior appointment of a health care agent in multivariate analysis. CONCLUSIONS: Differences in health care proxy completion rates across white, African American, and Hispanic elderly individuals in this New York City population seem to be related to potentially reversible barriers such as lack of knowledge and the perceived irrelevance of advance directives in the setting of involved family. Enhanced educational efforts of both health care personnel and patients could increase the rate of formal health care proxy appointment. PMID- 9855389 TI - Alveolar echinococcosis of the liver: sequelae of chronic inferior vena cava obstructions in the hepatic segment. AB - BACKGROUND: The clinical pattern and long-term course of chronic inferior vena cava (IVC) obstructions are variable and depend on the underlying cause, the segment involved, and the extension of secondary thrombosis. Pertinent data on IVC obstructions in well-defined series of patients are lacking. We report the sequelae of chronic IVC obstructions in the hepatic segment in 11 consecutive patients derived from a cohort of 104 patients with alveolar echinococcosis of the liver. METHODS: Based on the results of computed tomography scans, 11 patients (7 men, 4 women; mean age, 53.4 years) with IVC obstructions were selected from an ongoing prospective long-term chemotherapy trial comprising 104 patients with alveolar echinococcosis studied at yearly intervals according to a protocol. Obstruction of the IVC in the 11 patients existed for a mean duration of 8.6 years. In these patients, magnetic resonance imaging was performed to assess the morphologic features and extension of the IVC obstruction, as well as the collateral venous pathways. Patency and valvular function of the femoropopliteal veins were analyzed by color-coded duplex ultrasonography. RESULTS: Total occlusions of the IVC were evident in 8 patients (73%) and subtotal stenoses in 3 patients (27%). Only 4 patients (36%) exhibited signs and symptoms of chronic venous insufficiency of the lower extremities; 2 (18%) of the 4 had a history of swelling in the lower extremity. Seven patients (64%) had no lower extremity symptoms. One patient had a history of pulmonary embolism. Abdominal collateral veins were documented in 5 patients (45%) by using magnetic resonance imaging; however, they were clinically evident in only 3 patients (27%). In the 8 patients with IVC occlusion, thrombosis ended at the confluence of the hepatic veins. Obstruction of the IVC was limited to the hepatic segment in 2 patients (18%) and extended to the distal IVC or the iliofemoral veins in 6 patients (54%). Chronic venous insufficiency was present only if the femoropopliteal veins had been involved in the thrombotic process, showing residual venous obstruction, valvular incompetence, or both. Bilateral renal vein thrombosis with moderate proteinuria was observed in 2 patients (18%). The main collateral drainage was achieved through the ascending lumbar, azygos, and hemiazygos veins. CONCLUSIONS: In patients with alveolar echinococcosis, obstruction of the IVC in the hepatic segment often develops asymptomatically and rarely leads to the impairment of renal function. The collateral circulation fully compensates for obstruction of the IVC. Thrombotic involvement and valvular incompetence of the femoropopliteal veins seems to determine the development of chronic venous insufficiency of the lower extremities. PMID- 9855390 TI - The decreased serum urea nitrogen-creatinine ratio. AB - Sometimes, readily available laboratory data can provide valuable hidden information. Such is the case with the serum urea nitrogen-creatinine ratio. A normal ratio is 8:1 to 10:1. It is well known that an elevated ratio is seen in cases of prerenal or postrenal uremia. Less appreciated are the diagnostic possibilities suggested by a decreased serum urea nitrogen-creatinine ratio (ie, <8). Several clinical circumstances can lead to a decreased serum urea nitrogen creatinine ratio. PMID- 9855391 TI - Physician-assisted suicide. PMID- 9855392 TI - Differences in treatment and outcome for women with acute myocardial infarction. PMID- 9855393 TI - Cost-conscious cough control. PMID- 9855394 TI - Molecular epidemiology of dengue virus--the time for big science. PMID- 9855395 TI - Short communication: dengue serotype 2 subtype III ('Jamaica') epidemic in Santa Cruz, Bolivia. AB - BACKGROUND: During the past decade, dengue and dengue haemorrhagic fever (DHF) have become a public health problem in various Latin American countries. Indications of increased dengue cases in the city of Santa Cruz, Bolivia, early in 1997 were promptly investigated. METHODS: We conducted a sample sero-survey in one district of the city. Levels of antidengue IgM were determined and genetic analysis was performed on virus isolates. RESULTS: IgM antibodies were detected in 6.5% (95% CI: 3.4%-9.6%) of adults (over 15 years old) and 5.1% (2.0%-8.2%) of children (5-7 years old). Dengue virus serotype 2 subtype III ('Jamaica') was isolated. CONCLUSIONS: The estimated attack rates are compatible with a dengue epidemic in Santa Cruz. Isolation of dengue-2 'Jamaica' virus documents the further spread of this subtype from the Caribbean via Brazil into South America. Increased DHF preparedness seems mandatory. PMID- 9855396 TI - Changing clinical and biological manifestations of dengue during the dengue-2 epidemic in French Polynesia in 1996/97--description and analysis in a prospective study. AB - In August 1996 dengue-2 virus was detected in French Polynesia for the first time since 1976. A prospective study was conducted from November 1996 to April 1997. Each time one of 7 physicians suspected dengue, the patient was enrolled and epidemiological, clinical and biological data were recorded. Dengue diagnosis was confirmed by virus isolation and IgM detection. The aims of this study were to find clinical and biological predictive factors constituting a specific profile of dengue (DF) and dengue haemorrhagic fever (DHF/DSS) and to assess the possibility of diagnosing dengue at primary health care level using clinical criteria and basic laboratory parameters. Of 298 clinically suspect cases, 196 (66%) were confirmed as dengue. The association of macular rash, pruritus, low platelet count and leukopenia was statistically predictive of dengue but not clinically, since these four signs occur in many other viral infections. As the prevalence of clinical and biological manifestations varied over time in our study, a specific profile useful for dengue diagnosis cannot be defined. With six cases of DHF, the morbidity of this dengue-2 outbreak was very low despite the sequential infection scheme DEN-3/DEN-2. The clinical expression of dengue could depend on a specific virus strain circulating in a specific population in a particular place, with varying virulence over time. PMID- 9855397 TI - Possible dengue sequential infection: dengue spread in a neighbourhood during the 1996/97 dengue-2 epidemic in French Polynesia. AB - A DEN-2 epidemic occurred in French Polynesia from August 1996 to April 1997 after 7 years of DEN-3 circulation. The susceptible population constituted all expatriates and Polynesians under 21. In August 1996, two successive DEN-2 cases occurred in Teroma, a Tahitian neighbourhood close to the international airport of Tahiti. A serological prospective study of persons < 21 years living in Teroma was conducted. The study population was bled in September 1996, October 1996 and June 1997. Analysis of dengue spread in Teroma confirmed that dengue transmission occurs primarily in the house, thus vector control campaigns should incorporate focal insecticide spraying and systematic daily use of insecticide in houses. The evolution in time of the disease demonstrated that among a susceptible population, prevalence and incidence rates are related to the time of exposure, and consequently to age. Comparison of dengue incidence or dengue prevalence between populations therefore requires adjusted age rates. Most studies did not adjust for age, leading to the conclusion that DHF is more frequent during secondary than during primary dengue infection. Prospective studies taking into account the time of dengue exposure are necessary to confirm the sequential infection hypothesis. PMID- 9855398 TI - Yellow fever outbreak in Kaffrine, Senegal 1996: epidemiological and entomological findings. AB - In November 1996 a yellow fever (YF) outbreak occurred near Kaffrine in the central part of Senegal. Thirty-six deaths were notified, all children under 15 years of age. The YF diagnosis was confirmed by MAC-ELISA or by virus isolation. The immune status against YF virus of a sample population of 449 individuals was determined, and 31 confirmed cases and 69 asymptomatic cases were reported. Distribution of YF cases and incidence rate decreased with age, while the attack rate was stable in all age groups. Larva indices were high and Aedes aegypti was common in all villages, causing man-to-man transmission. The greatest risk of YF disease was lack of immunity, especially in individuals <20 years of age. The outbreak was rapidly controlled by an emergency immunization campaign. YF epidemics occurred in Senegal over two consecutive years. The last outbreak reached the main road to Dakar and the risk of spread to urban areas has increased. PMID- 9855399 TI - Outbreak of Crimean-Congo haemorrhagic fever in Quetta, Pakistan: contact tracing and risk assessment. AB - In December 1994 in a private hospital in Quetta, Pakistan, 3 health-workers contracted Crimean-Congo haemorrhagic fever (CCHF) after surgery on a bleeding patient who later died. We conducted a retrospective study to determine transmission risks among contacts. Fifty contacts gave blood for antibody tests and answered questions about exposure. Two of four people exposed percutaneously and one of five with cutaneous exposure contracted CCHE The person with cutaneous exposure was a surgeon who tore his glove during surgery and noted blood on his hand but no cut. There were no anti-CCHF antibodies or CCHF cases among persons whose skin came into contact with body fluids other than blood (0/4), who had skin-to-skin contact (0/16) with patients or were physically close to them (0/21). Three index case relatives reported that although 10 family members had cutaneous exposure, none developed CCHF. The family refused blood tests. CCHF transmission in resource-constrained settings can be limited by focusing on avoiding health worker contact with blood. PMID- 9855400 TI - Survey among survivors of the 1995 Ebola epidemic in Kikwit, Democratic Republic of Congo: their feelings and experiences. AB - This study describes experiences of the survivors of the 1995 Ebola epidemic in Kikwit, Democratic Republic of Congo. Most of the survivors in our sample had cared for a sick family member before becoming ill themselves, and most had never heard of Ebola before they developed symptoms and therefore did not suspect that they were infected by the virus. Fear, denial and shame were their principal initial feelings. After release from hospital, survivors were abandoned by family or friends more often than they had expected. Belief in god was an important aid to all of them. Their most negative experiences were witnessing other people dying in the isolation ward of the Kikwit General Hospital, and the reluctance of hospital personnel to treat them. During Ebola outbreaks more attention should be given to the psychosocial implications of such an epidemic. Information campaigns should include antidiscrimination messages and more psychosocial support should be given to patients and their families. PMID- 9855401 TI - Review of varicella zoster seroepidemiology in India and Southeast Asia. AB - Varicella zoster virus (VZV) predominantly affects children in temperate countries, with near-universal seroconversion occurring by late childhood. However, in tropical regions, VZV infection is common in adolescents and adults. This review identifies age-related VZV seroprevalence patterns in a number of Asian countries which indicate that seroconversion in tropical countries occurs at a later age than in temperate countries. Seasonal and regional variations in acute disease within some Asian countries suggest that temperate climates might favour transmission of the varicella virus, with incidence peaking during cooler months and in cooler, more temperate regions. VZV infection is often more severe in adults than in children, suggesting that tropical countries may be at risk of greater morbidity and mortality as a result of later-age seroconversion. Susceptibility of pregnant women and their infants, and of people infected with HIV/AIDS is also cause for concern. Vaccination may be beneficial in reducing the impact of VZV in Asian populations. PMID- 9855402 TI - Infection with group C rotavirus in a suburban community in Brazil. AB - Group C rotaviruses are associated with sporadic outbreaks of gastroenteritis worldwide. Age-specific seroprevalence of group C rotavirus antibodies was investigated in sera, randomly collected and representative of a suburban community in Brazil which had previously been screened for group A rotavirus antibodies. Antibody prevalence to group C rotavirus was low in children under 5 years and increased slowly with age to 36% seropositivity in adults, reflecting continuous exposure to primary infection in all age groups. This suggests a higher incidence of infection than disease might predict. Adult antibody prevalence was similar to that in other geographical settings. No obvious patterns of infection with group A and group C rotavirus were found within individuals, which suggests independent transmission. However, further epidemiological studies are required to understand group C rotavirus dynamics and possible interactions with group A rotavirus transmission and immunity. PMID- 9855403 TI - Incidence and risk factors of diarrhoea in Dutch travellers: consequences for priorities in pre-travel health advice. AB - A cohort of 743 Dutch short-term travellers (1-6 weeks) to various (sub)tropical areas was studied to assess incidences of travellers' diarrhoea (TD) and risk factors to guide prevention policies. The occurrence of TD was ascertained retrospectively by questionnaire; independent risk factors were identified by logistic regression analysis. The overall attack rate (AR, 95% CI) of TD was 52% (49-56); 11% (9-14) reported two or more episodes. The overall incidence rate (IR) per 100 person weeks of travel (pwt) (95% CI) was 22 (20-24). IRs were highest for travellers to the Middle East (48, 33-71), lowest for South-east Asia (17, 15-20) and East Africa (18, 14-24) and intermediate for South America and West Africa (both 26, 19-36), Central America (29, 23-37) and the Indian subcontinent (32, 26-39). Compared to first episodes of TD, subsequent episodes were of longer duration and more frequently accompanied by faecal blood loss, abdominal cramps or systemic symptoms. After adjustment for travel duration and destination, independent risk factors (OR, 95% CI) for TD were recent treatment for gastrointestinal (GI) disorders (4.6, 1.2-17.2), history of GI surgery (3.9, 1.4-11.1) and, possibly, current use of medication reducing gastric acidity (6.9, 0.7-67.4). The risk was reduced for extensive travel experience (0.4, 0.3-0.7) and organized travel (0.7, 0.5-0.9). Regarding prevention and/or antibiotic self treatment of TD, priority should be given to travellers who may suffer major health or other consequences from TD and to those with pre-existing GI disorders, particularly when visiting a high or intermediate-risk area on individual journeys with limited travel experience. PMID- 9855404 TI - Epidemiology of soil-transmitted nematode infections in Ha Nam Province, Vietnam. AB - Baseline epidemiological data are presented from a parasitological survey conducted in Thuy Loi commune, Ha Nam province, Vietnam; a farming community where night soil is routinely used as fertilizer for crops. 177 households were visited and 543 individuals (aged 1-88 years) recruited to the study. Helminth infection intensity was assessed by Kato-Katz to determine the density of parasite eggs per gram of stool (epg). Ascaris lumbricoides, Trichuris trichiura and hookworm infections were the only species detected. 83% of individuals were infected with A. lumbricoides (mean epg = 11,971), 94% with T. trichiura (mean epg = 793) and 59% with hookworm (mean epg = 302). Age-dependent patterns of infection prevalence and intensity were similar for A. lumbricoides and T. trichiura, but markedly different for hookworm infection. Similarly, age dependency in the k-values for the three infections was due to covariance with the respective mean intensities with age rather than to independent age effects, with similar patterns for A. lumbricoides and T. trichiura, and a different pattern for hookworm. Three major conclusions can be drawn from the multiple species analyses: There is positive interaction between A. lumbricoides and T. trichiura infections; high-intensity A. lumbricoides infections are significantly associated with high-intensity T. trichiura infections; and there is positive interaction between these two species such that infection intensity of A. lumbricoides is higher in individuals with concurrent T. trichiura infection than in individuals without and vice versa. PMID- 9855405 TI - Discrepancy in plasma melarsoprol concentrations between HPLC and bioassay methods in patients with T. gambiense sleeping sickness indicates that melarsoprol is metabolized. AB - OBJECTIVE: With the use of a specific high-performance liquid chromatography (HPLC) method and a bioassay which determines trypanocidal activity, concentrations of melarsoprol were assessed in plasma, urine and cerebrospinal fluid (CSF) from 8 patients with late-stage Trypanosoma gambiense sleeping sickness. The aim was to unravel to what extent the bioassay codetermines biologically active metabolites of melarsoprol. METHODS: Subjects were given one dose of melarsoprol i.v. per day for 4 days (1.2, 2.4, 3.0-3.6, 3.0-3.6 mg per kg b.w., respectively). Plasma samples were obtained before the first melarsoprol injection, immediately after and at 1 h, 24 h and 5 days after the 4th injection. Urine was collected before melarsoprol therapy and at 24 h after the 4th injection. CSF samples were taken once before treatment and at 24 h after the 4th injection. RESULTS: HPLC analyses showed that plasma concentrations immediately after the 4th injection varied from 2200 to 15,900 nmol/l; dropping to 0-1800 nmol/l at 1 h; and to undetectable levels at 24 h. In urine small amounts of melarsoprol were recovered. Melarsoprol could not be detected in CSF by HPLC. Immediately after injection, bioassay analyses showed plasma concentrations of the same magnitude as HPLC assays but at 1 h they were 4-65-fold higher than the levels assessed by HPLC. Even 24 h and 5 days after the 4th injection there was significant but decreasing activity. Urine levels were 40-260-fold higher than the measured HPLC concentrations, whereas there was low but detectable activity in CSF. CONCLUSION: Results indicate that melarsoprol is rapidly eliminated from plasma. The significant trypanocidal activity determined by bioassay and simultaneous low or undetectable levels of melarsoprol assayed by HPLC indicate that the compound is transformed into metabolites with parasiticidal activity. PMID- 9855406 TI - Ivermectin influence on the mast cell activity in nodules of onchocerciasis patients. AB - Onchocercal nodules were stained immunohistochemically using antibodies specific for human mast cells and IgE to elucidate the localization and frequency of mast cells after a single oral dose of 150 microg/kg ivermectin. Tryptase-and chymase positive mast cells occurred predominantly in mixed inflammatory infiltrates and perivascularly, and never adhered to adult worms or microfilariae. Up to three days after ivermectin, mast cells and IgE-positive cells were markedly increased in the capsular area of nodules containing female worms with embryos and microfilariae compared to untreated nodules. In the centre of these nodules, around the adult Onchocerca volvulus, we found many tryptase-positive cells. More mast cells were IgE-positive than in untreated nodules, equalling the number of tryptase-positive mast cells. There was a clear correlation between the appearance of mast cells and the attacks on damaged microfilariae by eosinophils and macrophages and in the vicinity of adult worms by neutrophils that occur soon after ivermectin treatment. Onchocercomata harbouring female worms with oocytes only revealed, after all treatment intervals, the same mast cell numbers as untreated nodules. In conclusion, during the first three days after administration, ivermectin produces increased numbers of mast cells in nodules harbouring females with embryos and microfilariae, probably as part of an allergic reaction to the attacked microfilariae. Four to 19 days after ivermectin the number of mast cells in the entire nodule is no longer elevated. PMID- 9855407 TI - Obstetric care in southern Tanzania: does it reach those in need? AB - OBJECTIVE: To assess whether antenatal care achieves identification and timely referral of high-risk pregnancies in southern Tanzania. METHODS: We compared the risk profiles of pregnant women in general with those attending obstetric care and investigated the reasons for seeking care. The risk profile of inpatients was drawn up through interviews with maternity cases and analysis of their antenatal records at the regional referral hospital (n = 415); population-based data on the prevalence of specific risk factors were obtained from entries in antenatal care registers (n = 1630) and from literature. RESULTS: A significant risk selection towards obstetric referral level care was observed only for previous caesarean section (prevalence hospital 6.7%, all pregnancies 1.5%, P < 0.005) and for nulliparity (hospital 42.8%, all pregnancies 25.0%, P < 0.005). No significant differences were observed for other risk factors such as previous perinatal death, height < 150 cm, multiple gestation and breech presentation. Prevalence of the risk factors age > 34 years and grand multiparity was significantly lower among hospital users. Coverage of obstetric care was below 50% for all risk factors except previous caesarean section (91.5%). CONCLUSION: Despite pursuing the risk approach and very good coverage, antenatal care in Tanzania has only limited effect on extending obstetric care to high-risk mothers. A critical review of the present screening and counselling practices, including a more focused and client-centred application of risk assessment, is warranted. PMID- 9855408 TI - David Magrath Memorial lecture: Lessons learned from the development of poliovaccines. PMID- 9855409 TI - Vaccines, human experimentation, and ethics in evolutionary perspective. AB - Clinical testing of unlicensed biological products is woven into issues of ethics and technology that reflect the state of the art. Materials for human testing must be of best possible quality and content, and must be safe. Testing of vaccines demands fulfillment of three ethical principles established since World War II. Thus, autonomy (informed consent), beneficence (optimize benefit to risk, and do no harm), and justice (equitable distribution of benefits and burden among volunteer participants). Ethical and technical considerations are the driving force for regulation, by National Control Authorities, of both products and clinical testing. Regulation is facilitated by guidelines promulgated by the larger National Control Authorities, by the European Community, and by the World Health Organization. Sane and sensible regulation is best achieved by well trained and informed regulatory staffs and is optimized by their continuing hands on conduct of research. The difficulties of the past products have been diminished by increasing technical sophistication. The future bodies well for improved and new vaccines that will be based mainly on the established technologies of the past. New technologies will be approved slowly. PMID- 9855410 TI - Overview of current preclinical testing strategies for viral vaccines. PMID- 9855411 TI - Review of current preclinical testing strategies for bacterial vaccines. AB - A wide variety of bacterial vaccines is in various stages of preclinical and clinical development. These products range from whole killed or live attenuated bacterial organisms to purified proteins, peptides and plasmid DNA. Although preclinical strategies may be directed by a set of common guidelines focused on demonstrating safety and biological activity, the exact developmental scheme will depend on product-specific characteristics. In general, preclinical data should support the proposed clinical formulation and include detailed information on the source and quality of starting materials, manufacturing processes, characterization of bacterial seed stocks, potency, general safety, purity, and identity. Data describing product validation and testing may be appropriate depending on the type of product, e.g., genetic stability for recombinant constructs, details on inactivation or attenuation methods for organisms or toxins, demonstration of potency of combination products, and safety and toxicology studies of plasmid DNA vaccines or vaccines with novel adjuvants. The choice of dose, route, and formulation to be used clinically may be greatly affected by rigorous preclinical developmental strategies. PMID- 9855412 TI - A respiratory challenge model for infection with Bordetella pertussis: application in the assessment of pertussis vaccine potency and in defining the mechanism of protective immunity. AB - The evaluation of vaccines for human use usually requires the development of appropriate animal models and the definition of laboratory correlates of immunity. Traditionally whole cell pertussis vaccines have been controlled by using an active mouse protection test, which measures protection following intracerebral challenge with Bordetella pertussis. However, this test is unsuitable for assessing the potency of the new generation acellular pertussis vaccines. In the present study we demonstrate that a murine respiratory challenge model for infection with B. pertussis is suitable for assessing the potency of acellular and whole cell pertussis vaccines. To allow standardization of different vaccines we have expressed the area under the clearance curve for immunized mice as a ratio of that for non-immunized controls to obtain a potency index. A comparison of estimated vaccine efficacy in children with potency in the murine model results in a highly significant correlation (r = 0.976, p < 0.001). Furthermore, we have used this model to define the protective mechanism of immunity against respiratory infection with B. pertussis and demonstrate a requirement for both specific T and B cells. In accordance with studies in humans, no clear relationship was observed between monotypic serum antibody responses against the putative protective antigens of B. pertussis and protection. In contrast, the most potent protection was observed when the T cell response is polarized to the Th1 subtype. PMID- 9855413 TI - DNA vaccines. AB - Immunization with plasmid DNA encoding antigenic proteins elicits both antibody and cell-mediated immune responses. This method of producing the protein antigens of interest directly in host cells can provide appropriate tertiary structure for the induction of conformationally specific antibodies, and also facilitates the induction of cellular immune responses. DNA immunization has provided effective protective immunity in various animal models. The immune responses induced by DNA vaccines may in some instances be preferable to those produced by immunization using conventional methods. DNA vaccination appears to be applicable to a variety of pathogens and is a useful method of raising immune responses. Thus this approach to vaccination has the potential to be a successful method of rapidly screening for antigens capable of inducing protective immunity, and of inducing protective immunity against pathogens of clinical importance. PMID- 9855414 TI - A phased approach to clinical testing: criteria for progressing from Phase I to Phase II to Phase III studies. AB - The overall intent of clinical testing is to establish, in a series of phased studies, the clinical tolerance and acceptable "safety" of the candidate vaccine, as well as the type, level and persistence of the immune response after its inoculation, to a representative target population, according to a convenient administration schedule. The final stages involve the direct or indirect demonstration of protective efficacy, if possible in the population(s) for which the vaccine is intended. In addition, consistency of production must be demonstrated. At all these stages, the amount of prior information from preclinical and other studies affects and informs the objectives and design of subsequent studies. Progression from one testing phase to the next is dependent upon attaining the pre-set objectives of each series of studies. The precise objectives to be met will be decided on a case-by-case basis. The earliest assessments in humans (Phase I) involve evaluation of short-term clinical tolerance as measured by local and general reactogenicity, and gross assessments of immunogenicity, in a small number of highly selected individuals in an idealised situation. The selection of "optimal" dose and schedule are the result of further dose-ranging investigations (Phase II), involving more volunteers, with longer, more detailed follow-up assessments. It is at this stage that the accumulated evidence on its immunogenicity profile should be sufficient to assess whether or not the vaccine is worthy of further development. The next level of investigation (Phase III) aims to measure with greater precision the vaccine protective efficacy in the intended target population(s) by comparison of infection and/or disease attack rates in vaccine and placebo recipients. In consistency studies different production lots, manufactured at commercial scale, are tested to demonstrate consistency of manufacture. Additional bridging studies to establish similarity of lots at different production scales, or studies of the duration of the immunity conferred, are conducted in parallel with the progression of the studies in the different phases mentioned above. These latter types of studies are usually carried out concurrently with Phase III studies. This progression continues into the post-marketing period (Phase IV) with surveillance of long term efficacy and observational studies of possible rare adverse events to establish "safety" with more confidence. This paper examines, in general, the aims and designs of studies in each phase as an introduction to the more specific publications that follow. PMID- 9855415 TI - Natural history and pathogenesis as they affect clinical trials. AB - Careful observations on the natural history of infectious diseases have been the inspiration behind many successful vaccines, including Jenner's successful demonstration of protection against smallpox. Many "experiments of nature" provide powerful data on the key components of protective immunity. An understanding of the basis of protective immunity in absolute terms or, more realistically, through the proxy of surrogates of immunity, lies at the heart of clinical trials of vaccines. An understanding of infectious disease pathogenesis is also critical in defining and distinguishing immunisation strategies that aim to prevent infection (e.g. measles vaccine) as distinct from prevention of disease (e.g. tetanus toxoid). Coupled to classical epidemiology and the careful analysis of clinical data, the tools of molecular and cell biology have revolutionised the tools available for vaccine research, including clinical trials. Molecular techniques have provided tools for investigating the population structure of pathogens. The importance of population diversity and its associated antigenic variation is a key factor in designing and carrying out clinical trials of vaccines. PMID- 9855416 TI - Regulatory aspects of clinical trials with vaccines. PMID- 9855417 TI - Epidemiological factors in clinical trial design. AB - Randomised, double-blind, placebo-controlled clinical trials are human experiments in which the process of randomisation and blinded observation greatly simplify the interpretation of the results. However, epidemiological issues when properly addressed allow the optimal preparation of the study with anticipation of factors to be included in the analysis which are independent of the randomisation process but can affect the results, for example, the effect of age, dose, time since vaccination and exposure to other diseases. Alternative methods of estimating vaccine efficacy may be necessary in some settings, and such methods may have limitations which must be acknowledged in interpretation. This paper reviews some recently published studies of vaccine efficacy for the application of epidemiological principles in design and analysis. PMID- 9855418 TI - Clinical evaluation of new pneumococcal vaccines: the Finnish approach. AB - Clinical evaluation of a new vaccine should be based on information about the epidemiology and pathogenesis of the infection to be prevented, the protective immunity to this infection, and the immunological properties of the vaccine to be evaluated. In this review we describe one approach to evaluate a new vaccine, using our experience with pneumococcal conjugates as an example. Information on the epidemiology, pathogenesis and host responses was collected mainly during 1987-1992, improvement in diagnostic methods was carried out during 1990-1995, and the immunogenicity of vaccine candidates was tested during 1992-1995. Based on these factors, a randomized, controlled, blinded efficacy trial of two pneumococcal conjugate vaccines against acute otitis media was started in Finland in 1995. Enrolment of 2,497 infants has now been completed by April 1997, and the follow-up phase will continue until 1999. PMID- 9855419 TI - Assessment of human CD4+ and CD8+ T lymphocyte responses in experimental viral vaccine studies. AB - The traditional in vitro correlate of immunological protection is the induction by a vaccine of neutralizing antibodies against the virus. It was formerly assumed that protection induced by a vaccine was solely due to neutralizing antibodies. Neutralizing antibodies are potent in the prevention of certain diseases, but virus-specific CD4+ T helper cells aid in the proliferation of specific antigen-triggered B cells to make antibodies. CD8+ T cells are responsible for eliminating virus-infected cells during viral illness, and may act as a second line of defence by becoming activated and eliminating any cells that become infected despite the presence of neutralizing antibodies, for example because of a large challenge dose or antigenic variation at the antibody combining sites. We will briefly review our approaches for measuring human virus specific CD4+ and CD8+ T cell responses to experimental vaccines. It is critically important to have sensitive, quality controlled assays, including positive controls. There are many potential variables in human T cell assays and pitfalls, which usually result in negative CTL results. Uninterpretable data are to be expected unless adequate preliminary testing has been done to establish sensitive, specific and controlled human antigen specific CD4+ and CD8+ T cell assays. PMID- 9855420 TI - B cell responses and immune memory. PMID- 9855421 TI - Immunological issues in vaccine trials: T-cell responses. AB - T-cell responses are both extremely diverse and dependent on the MHC of the immunized (or infected) individual. Apart from T-cell proliferation assays, the most informative functional T-cell assays are still difficult to perform. Antibody measurements provide a very indirect assessment of the helper arm of the T-cell response. On the other hand, measuring cytolytic T cells (CTL) remains a difficult task, which has precluded the evaluation of CTL responses in vaccine efficacy trials. Accordingly, even though there are reasons to suspect that CTL are important to clear certain infections and to vaccinate against certain diseases, particularly chronic viral infections such as that caused infection by HIV, the data to support these claims are largely missing in humans. Improving and automating CTL assays would have a significant impact on vaccine design. The Immunoscope technology is a PCR based approach which describes the T cell repertoire by several thousands of measurements. This allows the detection of clonal expansions and to evaluate the oligoclonality of pathological T cell infiltrates in humans. In the mouse, it has allowed us to establish the concept of public T-cell responses which are recurrent in individual animals sharing the same MHC. This concept can occasionally apply to humans since we found a public T cell expansion in DR2a patients at the onset of multiple sclerosis. Single chain class I MHC molecules have been produced, purified, homogeneously loaded with an antigenic peptide, and coated on to beads. This formulation appears to be efficient for induction of primary CTL in vitro . A similar approach can be used to purify peptide specific T cells, and its coupling with the Immunoscope technology is being considered. The potential of these new approaches for T-cell analyses will be discussed. PMID- 9855422 TI - Immunisation and the maturation of infant immune responses. AB - Maturation of the adaptive immune system occurs early in development and by 14 weeks gestation the developing foetus has circulating differentiated T cells and B cells capable of responding to antigen. While the immune system is capable of responding appropriately at birth to protein antigens, its capacity to respond to carbohydrates is limited. A consequence of this is the success of bacteria with carbohydrate capsules in causing invasive paediatric infections. Vaccines containing purified carbohydrates alone are thus limited in their immunogenicity and fail to provide protection for those most at risk. Conjugate vaccine technology, where a carbohydrate antigen is chemically coupled to a protein carrier, has overcome the limitation of carbohydrates as vaccine antigens. The first such vaccine to enjoy widespread use, the Haemophilus influenzae type b conjugate vaccines, have met with enormous success and have almost eradicated invasive Hib disease in those countries where their use in infancy is routine. Conjugate technology is thus being applied to a number of other vaccines in development, including Neisseria meningitidis groups A and C and Streptococcus pneumoniae vaccines. PMID- 9855423 TI - Maternal immunity and infant responses to immunization: factors influencing infant responses. AB - The presence of maternally-derived antibodies at the time of immunization is known to interfere frequently with active immunization, with variable levels of clinical significance. Deciphering the rules as the basis of such inhibitory effects on infant vaccine responses would certainly contribute to the development of vaccination strategies for early life. These questions were addressed in murine neonatal or early life immunization models using various antigens (measles, tetanus, RSV) and antigen-presentation systems (peptides, proteins, live attenuated vaccines, live recombinant vectors or DNA plasmids) in the absence or presence of maternal antibodies. Factors identified as crucial determinants of maternal antibody-mediated effects on both live and non-live vaccines include the relative amount of maternal antibodies and of vaccine antigens present at immunization, antigenic conformation, epitope specificity and the distinct influence on B-cell and T-cell vaccine responses. PMID- 9855424 TI - Mucosal responses to parenteral and mucosal vaccines. AB - Most human pathogens are acquired through mucosal portals of entry, and replicate in the mucosal tissues. Subsequently, the infecting agent may invade the blood stream and produce disease at distant systemic sites. However, a large number of pathogenic organisms are limited to development of disease only at the site of initial mucosal replication. Studies carried out with naturally acquired infections and mucosally delivered vaccines have provided strong evidence for the existence of a common mucosal immune system in the organized lymphoid follicles in respiratory and intestinal epithelium, and in the mucosa of genital tract, mammary glands, conjunctiva, upper airways, and the middle ear cavity. Mucosal application of live attenuated oral poliovaccine (OPV), rubella virus vaccine (RA 27/3), adenoviruses, influenza A virus, rotavirus, salmonella, and cholera vaccines have demonstrated consistent development of secretory IgA, serum antibody, and cellular immune responses. Mucosal immunization appears to result in preferential expression of several integrins and cell adhesion molecules associated with homing of lymphocytes to mucosal sites of immunization. Induction of mucosal immune responses often result in specific protection against reinfection challenge and against illness. Replicating agents introduced via the parenteral route also result in the development of mucosal responses and protection against systemic illness. Parenteral immunization with non-replicating agents often fails to induce specific mucosal responses. Such immunization, however, is quite effective in mounting high levels of serum antibody with development of protection against systemic illness. Parenteral vaccines, such as enhanced potency inactivated polio vaccine (eIPV), Haemophilus influenzae type B (HIB), hepatitis B virus (HBV), and other non-mucosal vaccines, have been highly effective in preventing systemic disease during subsequent exposure to natural infection. Recent evidence has shown that parenteral immunization can also be quite effective in inducing varying degrees of functional mucosal antibody responses as detected by ELISA and less frequently by neutralization. Systemic illnesses such as poliomyelitis and Haemophilus influenzae meningitis and community circulation of these agents has been eliminated or significantly limited in many parts of the world with the exclusive use of inactivated vaccines. Based on these observations, it is suggested that development of serum immunological responses are effective in the prevention of systemic disease regardless of the types of vaccines or route of their administration. However, induction of pathogen-specific antibody or cellular immunity at the mucosal sites is best elicited by mucosal application of the antigen. PMID- 9855425 TI - HIV-1 DNA based vaccine induces a CD8 mediated cross-clade CTL response. AB - Novel approaches for the generation of more effective vaccines for HIV-1 are of significant importance. In this report we analyse the immunogenicity and efficacy of a DNA vaccination approach in a chimpanzee model system. Three chimpanzees were vaccinated with DNA constructs which express the env, rev, gag and pol proteins. These animals developed specific cellular responses to these proteins, although the nature of the responses varied among the animals. We demonstrated that DNA vaccination led to a CD8 mediated killing of targets expressing the homologous clade B envelope as well as targets expressing heterologous clade E envelope. In addition seronegative individuals have been inoculated with a DNA construct which expresses the env, rev proteins. These studies serve as an important benchmark for the use of DNA vaccine technology for the production of protective immune responses. PMID- 9855426 TI - Standardization of acellular pertussis vaccine by assay of serum neutralizing antibodies to pertussis toxin (antitoxin): analogy with diphtheria toxoid. AB - Immunity conferred by vaccination with cellular pertussis vaccines and pertussis wanes so that adults are now the main reservoir of B. pertussis. Similar to diphtheria toxoid, acellular pertussis vaccines are suitable for vaccination of adults and it is probable that addition of pertussis toxoid to DT will be recommended for this age group. If this can be achieved, it is easy to predict that pertussis will be eliminated and the possibility of eradicating B. pertussis will become feasible. We propose that, despite the relative inaccuracy of correlating levels of antitoxin with individual immunity, a standardized assay and reference antiserum for pertussis toxoid will control acellular pertussis vaccines as was achieved with diphtheria toxoid. PMID- 9855427 TI - Standardization may suffice for licensure of conjugate vaccines. AB - Standardization schemes devised by Control Agencies have followed clinical trials of experimental vaccines. The wealth of information about the pathogenesis of and immunity to bacteria, whose surface polysaccharides are protective antigens, now permits standardization to predict the efficacy of polysaccharide-based vaccines. There has been tacit acceptance of this notion with the licensure of groups Y and W135 meningococcal vaccines and of many of the pneumococcal types whose frequency in patients was too low for statistical significance to be assigned for their clinical efficacy. In fact, this was also the case for licensure of polio virus type 2 vaccine. We can reliably measure the level of anti-polysaccharide antibodies for meningococci, pneumococci, GBS and the Vi of S. typhi. Haemophilus type b conjugates have been reliably standardized by physico-chemical assays. New conjugates, therefore, may be licensed by data provided by standardization without awaiting the results of costly and time-consuming efficacy trials. Adoption of this scientifically-based approach to licensure will hasten the implementation of new and more effective vaccines. PMID- 9855428 TI - Experimental challenge studies in the development of vaccines for infectious diseases. PMID- 9855429 TI - Immunogenicity assays and surrogate markers to predict vaccine efficacy. AB - Immunogenicity of vaccines is most often evaluated by measuring the antibody concentration before and after vaccination. The assessment of cellular responses is far more laborious and difficult to standardize. However, the antibody concentration is not the sole criterion because different vaccine formulations can induce antibodies of different fine specificity and functional activity. The quality of antibodies can be measured in several ways; e.g. the information on isotype distribution, avidity and bactericidal, opsonizing and neutralizing capacity can be useful. As an example, the use of these surrogate assays for evaluation of Haemophilus influenzae type b (Hib) conjugate vaccines is described. The same assays can most probably be used to assess the potency of other conjugate vaccines that are now in phase I-III trials. PMID- 9855430 TI - Bridging studies. AB - The duplication of usually large-scale efficacy trials to generalise the clinical database of a new vaccine to other populations can be avoided by bridging studies. Immunogenicity data can easily be used to extrapolate efficacy results when the immune response correlates with vaccine induced immunity. In the absence of such a correlate of protection, the bridging process will be more controversial. Previous experiences should be valuable in identifying the factors, either ethnic or vaccine-related that may have an influence on vaccine efficacy. The probability that such a factor is associated with a clinically significant difference in efficacy is critical to the decision to conduct a bridging study. Bridging studies are designed to demonstrate equivalent immunogenicity i.e. exclude a clinically significant difference in the immune response between the population in whom efficacy was shown and the population to whom those efficacy results are extrapolated. PMID- 9855431 TI - Long-term efficacy of pertussis vaccines in Italy. PROPER Study Working Group. PMID- 9855432 TI - Measuring protection: efficacy versus effectiveness. AB - In the clinical development of a vaccine, an efficacy study asks the question, "Does the vaccine work?" In contrast, an effectiveness study asks the question "Does vaccination help people?". In general, vaccine development proceeds from a study of immunogenicity to a randomized controlled trial that determines vaccine efficacy under ideal conditions. Efficacy studies, however, have several limitations. In an immunogenicity study, when a vaccine is given according to different schedules, the object of the study is not the vaccine itself but the schedules; i.e., what is important is not the "relative immunogenicity" of the vaccine, but which schedule is more protective given the occurrence of the disease that is to be prevented. Furthermore, a clinical trial of vaccine efficacy is unable to predict accurately the level of protection that will be achieved in public health practice. Vaccination effectiveness can be evaluated in a prospective clinical trial, although few such studies have been undertaken. Effectiveness is usually assessed retrospectively, sometimes using a screening test, but more often in a case-control or cohort study. In these studies, rigorous risk adjustment is necessary to ensure the comparability of study populations. Retrospective studies also provide a means for assessing serious but rare vaccine-associated adverse events, an undertaking often needed to maintain public confidence in vaccination programmes. Many vaccines currently under development will be replacement rather than new vaccines, and they are unlikely to be evaluated in traditional efficacy trials. In future years, effectiveness studies, some of them using large administrative databases, will become increasingly important features of vaccine development and the formulation of public policy for immunization. PMID- 9855433 TI - Monitoring and safety assessment in Phase I to III clinical trials. AB - Safety issues play an important role for registration of new vaccines due to growing awareness of regulators and growing safety concerns of the general public, whether they are justified or not. Safety of a vaccine implies much more than merely monitoring and reporting adverse events, but relies on a holistic view including control of manufacturing, surveillance of possible long term effects, epidemiological data, the vaccination schedule and signals from post marketing surveillance. Vaccine safety has scientific and organisational implications. To achieve a balanced risk-to-benefit evaluation the input of a central safety department is recommended from planning of a clinical development to post-marketing phase. The concept of data evaluation and data assessment should be medically driven in close cooperation with the biometricians. An integrated safety plan with standardized procedures improves the likelihood of detecting a realistic AE-profile, prior to marketing a vaccine. The unique opportunity of a controlled clinical trial should be taken advantage of by optimizing both the organisation and the scientific methods. Further development of methodology of safety monitoring and assessment should be given the same importance as proving efficacy and complying with the regulatory requirements. PMID- 9855434 TI - Measuring protection; a case study of pertussis vaccines--Swedish Trial II: secondary non-randomized comparisons between two schedules of infant vaccination. AB - In a double-blind trial two-, three- and five-component acellular vaccines were compared to a British whole-cell vaccine: in areas using three doses at three, five and 12 months of age (3-5-12 schedule), 72,698 children and in areas using a two, four and six months schedule (2-4-6 schedule), 10,194 children were evenly randomized to the four groups. The background incidence of pertussis was higher in the 3-5-12 schedule areas than in the 2-4-6 schedule areas; in spite of this, the point estimates of the relative risks for the 3-5-12 schedule versus the 2-4 6 schedule were close to or below one for the multicomponent acellular and the whole-cell vaccine groups, indicating a lower overall risk of pertussis when the third dose was delayed. The risk of whooping cough according to parents was lowest for the five-component and whole-cell vaccine groups in both schedules. The delayed third dose elicited booster responses for filamentous haemagglutinin but not for the other pertussis antigens. For highly efficacious pertussis vaccines two doses in infancy followed by a third dose in the second year of life may be recommended. PMID- 9855435 TI - Measurement of human serum IgG antibodies or a surrogate is sufficient to standardize (predict efficacy) vaccines. PMID- 9855436 TI - Mathematical modelling of effectiveness. AB - The effectiveness of a vaccine can be broadly defined as a practical measure of its costs and benefits when used in a community setting. What then is the relationship between a vaccine's efficacy and its effectiveness? Effectiveness certainly includes efficacy, but also encompasses secondary effects of vaccination, long-term efficacy of a vaccine (beyond that measured in conventional efficacy trials), the impact of vaccinating against one strain upon cross-reacting infections, costs of vaccination programmes and logistic considerations for vaccine delivery. This paper considers some of the biological determinants of effectiveness namely; secondary effects, long-term efficacy and competitive interactions amongst strains. Mathematical models of the transmission dynamics of infection and its disruption by vaccination are used as tools to help unravel some of the complexities that may arise when making interventions in non linear biological systems. PMID- 9855437 TI - Safety assessment post-licensure. AB - The objectives of post-licensure surveillance are to identify rare or novel adverse events, estimate their rate of occurrence and distinguish those that are causally related to vaccination. Evidence of causality is based on biological plausibility supported by laboratory evidence of vaccine involvement and/or a statistically significant excess of events in the post-vaccination period. Passive reporting systems have methodological limitations, particularly for ascertaining reliable adverse event rates and investigating causal relationships. In Canada, an active surveillance system (IMPACT) has been established in sentinel paediatric hospitals where nurses scrutinise all admissions for conditions conforming to a defined set of adverse events. While ascertainment of events resulting in hospital admission should be complete, causality is difficult to assess without information on the incidence of such events outside the post vaccination risk period. The use of computerised data bases which link clinical events with immunisation records have allowed the risk of events such as convulsions attributable to DTP, MMR and Hib vaccines to be defined. The record linkage method promises to be a useful method for routine surveillance of vaccine safety. PMID- 9855438 TI - Clinical safety evaluation of combination vaccines. AB - The development plan for new combination vaccines should include clinical studies designed to provide an adequate safety database (as well as efficacy data) to support licensure. Ideally, randomized studies to compare the safety of the combination vaccine with the separately administered components (or already licensed combinations of components) should be performed. For new combination vaccines having components with proven efficacy, comparative immunogenicity data may provide a sufficient basis to support efficacy, precluding the need for a large efficacy study. In the absence of the safety database that would have been derived from such an efficacy study, it is important to conduct a comparative safety study with a sample size suitable for the evaluation of less common adverse events. For large safety trials, a simplified design where only a subset of subjects is assessed in detail for the more common adverse events may be appropriate. PMID- 9855440 TI - From immunisation research to policy and implementation. PMID- 9855439 TI - Development and efficacy assessment of combination vaccines, with emphasis on acellular pertussis. AB - The number of vaccines suitable for infant immunization is increasing, leading to a need to co-administer them in combination or combos. The recent demonstration of high efficacy against whooping cough using DTPa vaccines makes it desirable to administer combos such as DTPaHBIPVHib. For all vaccines included, surrogate markers for protection exist, except for pertussis. The field efficacy studies performed with DTPa vaccines did not allow the establishment of a correlate for protection. We propose to use detailed serology and cell-mediated immunity markers obtained from infant immunization studies, as well as a mouse intranasal challenge model, to monitor the immune responses induced by DTPa-combos. PMID- 9855441 TI - Pharmacologic therapy of idiopathic pulmonary fibrosis. PMID- 9855442 TI - Single-field total lymphoid irradiation in the treatment of refractory rejection after heart transplantation. AB - Nine heart transplant recipients were treated with single-field total lymphoid irradiation (TLI) for early (<1 year) or late (>1 year) rejection that was refractory to multiple regimens of immunosuppressive therapy. For patients with early rejection (n = 6), the rejection frequency (rejections/patient/month) decreased from pre-TLI of 1.63 to post-TLI of .02 (p < .001), and for patients with late rejection (n = 3), the rejection frequency decreased from pre-TLI of .23 to post-TLI of .05 (p < .02). The reduced rejection frequencies have been maintained for a mean follow-up of 28.6 (8 to 78) months, and adverse events during or late after TLI were uncommon. Single-field TLI is a safe and effective technique in the management of refractory rejection early or late after heart transplantation. PMID- 9855443 TI - Diastolic dysfunction coincides with early mild transplant rejection: in situ measurements in a heterotopic rat heart transplant model. AB - BACKGROUND: Diastolic dysfunction seen in early clinical transplant rejection has been difficult to demonstrate in experimental rodent models because of the inability to make sensitive in situ measurements of systolic and diastolic functions. We have developed a heterotopic heart transplant model with Fisher 344 and ACI rats (without immunosuppression), where in situ measurements of diastolic and systolic functions were made sequentially (daily) by use of an implanted left ventricular balloon. METHODS: Syngeneic and allogeneic heterotopic heart transplants were performed. In situ function was determined by varying balloon volume to measure the developed pressure, the rates of pressure rise (+dp/dt) and pressure fall (-dp/dt), diastolic pressure-volume relationship, and the time constant of diastolic relaxation (tau). These results were compared with function measurements in transplanted hearts that were excised and perfused in a Langendorff mode (ex vivo) during the same posttransplantation period. RESULTS: Histologic examination revealed that at day 3 after transplantation, allografts showed mild lymphocytic infiltration indicative of mild or early rejection, and by day 5, there was severe rejection with myocyte necrosis. By day 3, the slope of the diastolic pressure-volume relationship (ie, left ventricular stiffness) was significantly higher in allografts as compared with isografts (436 +/- 96 vs 177 +/- 26 mm Hg/mL, p < .05). Similarly, tau was significantly longer in allografts by day 3 after transplantation. Developed pressure and +dp/dt became significantly lower in allografts beginning on day 6. Function measurements made in the isolated perfused ex vivo hearts yielded the same results at day 3 after transplantation as the in situ group of hearts. CONCLUSION: Using a chronically implanted left ventricular balloon, we have developed a heterotopic heart transplant model where sensitive measurements of systolic and diastolic functions can be made. With this preparation, the early changes in the diastolic dysfunction seen clinically can be reproducibly detected. Thus this model may be useful to study mechanisms and interventions during early transplant rejection. PMID- 9855444 TI - Modified technique of abdominal heart transplantation in the rat. AB - BACKGROUND: A rapid, reproducible screening model is essential for evaluation of novel preservation regimens. This study describes a modification of the abdominal rat heart transplantation model reducing anastomosis time and allowing quantitative assessment for 7 days. METHODS: Hearts, obtained from inbred Dark Agouti rats, were arrested and stored in cold colloid-free University of Wisconsin solution until transplantation. The Dark Agouti recipient underwent a left nephrectomy. The donor left common carotid artery was anastomosed to the recipient left renal artery with a "sleeve" anastomosis. The "cuffed" donor left pulmonary artery was inserted into the left renal vein. Study 1 examined continuing viability by daily palpation and morphologic study by examination of hematoxylin and eosin-stained sections on days 4 or 90. Study 2 examined quantitative assessment of cardiac function in the anesthetized recipient. The model was further modified by introducing an externalized, fluid-filled, balloon tipped catheter into the left ventricle. RESULTS: The new technique allowed vascular anastomoses to be completed in 5 to 12 minutes, minimizing rewarming of the graft. Most (25 of 28) grafts beat for 90 days, and 80% of these showed normal structure. There was evidence of myocyte damage or arteriosclerosis in 5 of 25 at 90 days and in 4 of 17 at 4 days. Cardiac function parameters were similar in consecutive runs and did not change between days 1 and 7. CONCLUSION: This abdominal rat heart transplant model is quick and easy to perform, minimizes warm ischemia, and is suitable for both short- and long-term studies. Quantitative parameters, assessed by use of an in situ intraventricular balloon tipped catheter, are reproducible and maintained for 7 days. PMID- 9855445 TI - The bicaval anastomosis technique for orthotopic heart transplantation yields better atrial function than the standard technique: an echocardiographic automatic boundary detection study. AB - BACKGROUND: Atrial function is an important determinant of cardiac performance. In patients who undergo operation by standard heart transplantation atrial enlargement, distortion of geometry and asynchronous contraction resulting from the donor/recipient atrial connections may affect atrial function. The bicaval anastomosis technique should be free from these limitations. METHODS: We used the echocardiographic automatic boundary detection technique to obtain on-line time/volume curves of right and left atria from patients who had undergone bicaval (n = 22) or standard (n = 27) heart transplantation and from 15 control subjects. Maximal, middiastolic, preatrial contraction, and minimal volumes of both atria were measured. Reservoir volume (defined as the difference between maximal and middiastolic atrial volumes); pump volume (defined as the difference between preatrial contraction and minimal atrial volumes); and conduit volume (defined as the difference between left ventricular stroke volume and the sum of reservoir and pump volumes) were derived for both atria. Atrial emptying fraction was calculated as the difference between maximal and minimal volumes divided by the maximal volume and expressed in percent and pump fraction as the pump volume divided by the sum of reservoir and pump volumes. Tricuspid and mitral regurgitation, evaluated by color-flow Doppler scanning, were considered significant when they were greater than grade 1. Atrial ejection force was calculated from mitral and tricuspid flow velocities at atrial contraction. RESULTS: In patients who had bicaval heart transplantation, both atria were smaller than in patients who underwent standard heart transplantation. With the bicaval technique right and left atrial emptying (right 45% +/- 9% vs 36% +/- 10%, p < .05; left 51% +/- 8% vs 39% +/- 8%, p < .001) and pump fractions (right 57% +/- 17% vs 19% +/- 13%, p < .001; left 45% +/- 28% vs 22% +/- 12%, p < .01) were greater than with the standard technique and similar to those in control subjects. Right atrial ejection force was significantly greater in bicaval (10.0 +/- 5.6 kdyne) than in standard heart transplantation (4.5 +/- 2.2 kdyne, p < .0001). Significant tricuspid or mitral regurgitation was rarely found in bicaval heart transplant recipients (3 and 1 of the 22 patients, respectively), although they were much more frequent after standard heart transplantation (13 and 8 of the 27 patients, respectively). CONCLUSIONS: Heart transplantation performed with the bicaval anastomosis technique determines smaller atrial volumes, yields better right and left atrial function and fewer atrioventricular valve regurgitation than the standard technique. PMID- 9855446 TI - Safety of influenza vaccine in heart transplant recipients. AB - BACKGROUND: Influenza vaccine is recommended for heart transplant recipients, but its administration is often deferred because of anecdotal reports of rejection associated with the vaccine. We evaluated the safety of influenza vaccine in a group of stable heart transplant recipients over a 2-year period. METHODS: During the 1993 to 1994 influenza season, stable heart transplant recipients who had undergone transplantation a minimum of 1 year before study entry were randomized to vaccination with a single dose of influenza vaccine versus no vaccination. Routine endomyocardial biopsies and postvaccination influenza serologic studies were performed between 2 and 6 weeks after enrollment/immunization. During the 1994 to 1995 season, patients were given 2 doses of influenza vaccine, separated by 3 weeks; endomyocardial biopsies and serologic studies were performed between 2 and 6 weeks after the second immunization or enrollment (if control subject). Biopsy results were evaluated with respect to vaccine response, immunosuppressive regimens, and patient demographics. RESULTS: Eighteen patients were enrolled in the single vaccine trial and 10 in the booster vaccine trial. Four of 14 vaccine recipients had biopsy specimens consistent with International Society for Heart and Lung Transplantation grades 2 to 3A as compared with 1 of 14 control subjects (grade 2) (p = .326). All episodes of rejection in the vaccine recipients were asymptomatic and responded to a single course of treatment. Rejection was unrelated to the time from transplantation, doses of immunosuppression, age, or number of doses of or response to vaccine. CONCLUSIONS: Influenza vaccine can be safely administered to most heart transplant recipients but may be associated with low-level histologic rejection. PMID- 9855447 TI - Normal short-term renal response to acute volume expansion in heart transplant recipients: a role for atrial natriuretic peptide? AB - BACKGROUND: The breakdown of blood pressure and body fluid homeostasis observed in heart transplant (Htx) recipients may partly be due, as in heart failure, to a blunted renal response to elevated atrial natriuretic peptide (ANP). METHOD: This possibility was addressed through determination of the relationship between ANP, the urinary cyclic guanosine monophosphate (cGMP), a biologic marker of ANP renal activity, and the early renal responses to 10 mL/kg isotonic saline infusion over 30 minutes in 8 control subjects and 8 matched Htx recipients. RESULTS: Urine flow, natriuresis, and urinary cGMP excretion increased similarly in both groups, resulting in elimination of, respectively, 1/2 and 2/3 of the sodium and the water load during the experiment that lasted 4 hours and 30 minutes. Plasma renin and aldosterone decreases were similar in both groups. Elevated ANP further increased in Htx after saline infusion (from 19.5 +/- 3.7 to 33.8 +/- 5.6 pmol/L, P < .001). Plasma cGMP paralleled ANP in both groups (r = 0.81; P < .001). Significant correlations were observed between plasma ANP and urinary cGMP excretion (r = 0.48, P < .025 and r = 0.43, P < .05 in Htx recipients and control subjects) and between plasma ANP and urinary sodium excretion (r = 0.64, P < .001 in Htx recipients). CONCLUSION: In spite of a relative renal hyporesponsiveness to the cardiac hormone, with higher plasma ANP being not associated with increased renal excretions in Htx recipients, ANP is likely to participate in the appropriate short-term renal response to acute volume expansion in Htx recipients. PMID- 9855449 TI - Body mass index as a predictor of survival in adults with cystic fibrosis referred for lung transplantation. AB - BACKGROUND: The timing of referral and listing for lung transplantation in adults with cystic fibrosis is influenced by many factors including pulmonary function, body mass index (BMI), sex, and patient and physician choice. This study aimed to analyze the effect of these variables on waiting list and postoperative mortality rates. In particular, low BMI is suggested to portend a poor outcome after transplantation. METHODS: All patients with cystic fibrosis referred to our institution (n = 92) between 1989 and 1996 were reviewed, and the effect on survival of BMI, sex, and other covariates was analyzed by use of Cox proportional hazards regression. RESULTS: Forty-five transplantations were undertaken with a mean waiting time of 226 days (range 1 to 678). Fifteen of the 62 listed patients died before transplantation with a mean time to death of 160 days (range 8 to 533). Fifteen patients died after transplantation. BMI at the time of listing predicted waiting list mortality (P < .05). Female sex tended to increase waiting list mortality rates, such that the combination of BMI less than 18 kg/m2, and female sex was associated with a 21% 1-year waiting list survival without transplantation. Age, forced expiratory volume in 1 second, sex, BMI, and date of transplantation did not predict postoperative survival. CONCLUSION: Patients with cystic fibrosis (particularly women) referred for lung transplantation with a BMI less than 18 kg/m2 are at high risk of death over the next 12 months. With this in mind, they should not be denied transplantation unduly while attempts are made to increase weight, especially because pretransplantation BMI does not influence posttransplantation survival. PMID- 9855448 TI - Prevention of bone loss after heart transplantation with antiresorptive therapy: a pilot study. AB - BACKGROUND: Heart transplantation, with its attendant glucocorticoid and cyclosporine therapy, has deleterious effects on the skeleton. We have previously reported rapid bone loss and high fracture rates (36% of patients) during the first year after heart transplantation. The bone loss was accompanied by declines in serum 1,25-dihydroxyvitamin D and osteocalcin levels and increased urinary excretion of markers of bone resorption (hydroxyproline, pyridinoline, and deoxypyridinoline). We therefore investigated whether bone loss could be prevented by bisphosphonates, agents that inhibit bone resorption. METHODS: Serial measurements of bone mineral density (BMD) and biochemical indexes of mineral metabolism were compared in 18 group A patients who received a single intravenous infusion of pamidronate (60 mg) within 2 weeks of heart transplantation, followed by 4 cycles of oral etidronate (400 mg daily for 14 days every 3 months) and oral calcitriol 0.25 microg daily, to those of 52 patients who previously underwent transplantation (group B) who did not receive antiresorptive therapy. Both groups received elemental calcium 1000 mg and vitamin D 400 IU daily. RESULTS: At 12 months after transplantation, there was virtually no lumbar spine bone loss in group A patients, whereas lumbar spine BMD had declined significantly in group B patients (0.2% +/- 0.9% vs 6.8% +/- 1.0%, respectively; P < .0001). Similarly, femoral neck BMD fell by 10.6% +/- 1.1% in group B patients and by only 2.7% +/- 1.4% in group A patients (P < .0001). Three incident vertebral fractures occurred in two group A patients, whereas 17 group B patients sustained 30 incident vertebral fractures, one hip fracture and three episodes of rib fractures (P < .02; test of proportions). With respect to markers of bone resorption, urinary deoxypyridinoline fell by 51% +/- 9% in group A patients and increased by 65% +/- 22% in group B patients by 3 months after transplantation (P < .0001). CONCLUSION: In summary, heart transplant recipients treated with bisphosphonates and replacement doses of calcitriol sustained less bone loss and fewer fractures than those treated with calcium and vitamin D. We conclude that bisphosphonate therapy, in conjunction with calcitriol, shows promise for prevention of transplantation-related osteoporosis. PMID- 9855450 TI - Exercise performance in cystic fibrosis before and after bilateral lung transplantation. AB - BACKGROUND: Lung transplantation improves pulmonary function and quality of life for patients with end-stage cystic fibrosis; however, a systematic evaluation of exercise performance in lung transplant recipients with cystic fibrosis has not been reported. METHODS: Ten patients with end-stage cystic fibrosis performed incremental exercise testing before and after bilateral lung transplantation; their results were compared with those of 10 age-similar healthy volunteers. Breath-by-breath measurements of gas exchange and ventilation were obtained, arterial blood was sampled each minute, and cardiac output determined at rest and peak exercise by radionuclide ventriculography. The arterial-venous O2 content difference was derived by the Fick principle. RESULTS: After transplantation, peak O2 uptake improved (31% +/- 3% vs 45% +/- 4% predicted, P = .03) but was still reduced versus normal (100% +/- 8% predicted, p < .0001). Exercise was limited by pulmonary mechanics in all patients before transplantation but in only 2 after transplantation. Compared with control subjects, the lactate threshold occurred early, both before and after transplantation. Peak exercise cardiac output and arterial O2 content were not different from normal, either before or after transplantation. In contrast, the peak exercise arterial-venous O2 content difference was markedly reduced before and after transplantation versus normal (7.1 +/- 1.2 and 9.3 +/- 0.9 vs 17.1 +/- 1.2 mL/dL, p < or = .0001 for each) and without significant improvement. CONCLUSIONS: Exercise performance in patients with end-stage cystic fibrosis improves after lung transplantation but remains well below normal. Reduced systemic O2 extraction is an important factor limiting exercise in patients with cystic fibrosis after transplantation and may also contribute to the exercise limit before transplantation. PMID- 9855451 TI - The nitric oxide donor, diethylamine NONOate, enhances preservation of the donor rat heart. AB - BACKGROUND: Ischemia/reperfusion injury to transplanted organs may be associated with loss of endothelial release of nitric oxide. The aim of this study was to determine whether supplementation of an extracellular-based cardioplegic solution in routine clinical use at our institution with nitric oxide (as diethylamine NONOate) enhanced poststorage functionality of an isolated working heart model. METHODS: Excised hearts were ligated to an aortic cannula and immediately perfused retrogradely with oxygenated Krebs solution at a hydrostatic pressure of 100 cm H2O at 37 degrees C. This preparation was then converted to a working system by switching the supply of perfusate from the aorta to a left atrial cannula at a filling pressure of 15 cm H2O. After a 1-minute stabilization period, baseline measurements of heart rate, aortic flow, coronary artery flow, and cardiac output were performed. Oxygenated cardioplegic solution (0.1 micromol/L), with or without NONOate, was then infused into the coronary circulation. Hearts were then stored in the same solutions for 6 or 12 hours at 2 degrees to 3 degrees C. The hearts were then remounted on the perfusion apparatus and reperfused as before, and hemodynamic measurements were repeated. Water content of the hearts were then determined. RESULTS: Addition of the nitric oxide donor significantly improved all hemodynamic parameters measured after 12 hours storage and aortic flow at 6 hours storage compared with the untreated control groups. There was no significant difference between the water contents of the NONOate-treated and control groups. CONCLUSIONS: The presence of the nitric oxide donor diethylamine NONOate was associated with significantly better preservation of coronary artery flow and cardiac function in the isolated rat heart after a 12 hour period of hypothermic storage and suggests a novel use for this family of compounds in the transplantation context. PMID- 9855452 TI - Prevention of rapid reperfusion-induced lung injury with prostaglandin E1 during the initial period of reperfusion. AB - We have found that the instantaneous restoration of blood flow causes acute dysfunction and massive edema in rat lungs after 4 hours of room temperature ischemia. This is associated with an early increase in pulmonary artery pressure (Ppa) and can be prevented by a stepwise increase in flow rate during the first 10 minutes of reperfusion. The objectives of this study were to determine whether rapid reperfusion causes lung injury after hypothermic preservation, and whether this injury can be attenuated by a short-course of prostaglandin E1 (PGE1). Rat lungs were flushed preserved with low-potassium dextran solution for 12 hours at 4 degrees C and randomly divided into three groups: (1) control (no PGE1); (2) PGE1 only in the flush solution; and (3) PGE1 in both flush solution and blood perfusate during the first 10 minutes of reperfusion. Postpreservation pulmonary function was assessed in an isolated rat lung reperfusion model developed previously. We found that rapid initiation of reperfusion led to significant pulmonary dysfunction, which was attenuated by a short-course of PGE1 in the blood perfusate. The addition of PGE1 to the flush solution alone did not have such an effect. Administration of PGE1 to the blood perfusate during the first 10 minutes resulted in significant lower Ppa and airway pressure and better gas exchange. There was a positive correlation between the peak Ppa during the first 10 minutes of reperfusion and the final shunt fraction. The physical forces generated by the rapid initiation of blood reperfusion appear to induce severe injury. The first 10 minutes of reperfusion seem to be a transition phase in which mechanical factors play an important role relating to ultimate post reperfusion lung function. A short course of PGE1 may be a useful maneuver to prevent rapid reperfusion-induced lung injury. PMID- 9855453 TI - Lung transplantation for pulmonary amyloidosis: a case report. PMID- 9855454 TI - Recurrent left-sided heart leiomyosarcoma: should heart transplantation be legitimate? AB - Leiomyosarcoma of the heart is an uncommon primary malignant tumor with poor postoperative survival that may be measured in months. A leiomyosarcoma of the left atrium was diagnosed in a 47-year-old man. Initial admission was for acute pulmonary edema requiring emergency surgery. The tumor involved the left atrial cavity, and a radical resection was performed. Six months later an isolated myxomatous recurrence was detected. Heart transplantation was then performed. The patient is in good health 20 months after operation with no evidence of residual disease or recurrence. The literature has been reviewed. Surgical resection is not an adequate treatment for leiomyosarcoma of the left atrium and early heart transplantation probably offers the only hope for these patients. PMID- 9855455 TI - Troglitazone, a new antidiabetic agent, decreases cyclosporine level. PMID- 9855456 TI - Osteoclast precursors as leukocytes: importance of the area code. PMID- 9855457 TI - Osteoclastogenesis inhibitory factor exhibits hypocalcemic effects in normal mice and in hypercalcemic nude mice carrying tumors associated with humoral hypercalcemia of malignancy. AB - Osteoclastogenesis inhibitory factor (OCIF) is a novel secreted protein that inhibits osteoclastogenesis both in vitro and in vivo. In this study, we examined the effects of OCIF on serum calcium (Ca) concentrations in normal mice and in hypercalcemic nude mice carrying tumors associated with humoral hypercalcemia of malignancy. In normal mice, a single intraperitoneal injection of OCIF reduced serum Ca levels in a dose-dependent manner. Significant decrease in serum Ca (by 1.6 +/- 0.3 mg/dL, n = 5) was observed 2 h after the injection of OCIF at 20 mg/kg and the hypocalcemic effect continued for up to 12 h. Serum phosphate (Pi) concentrations also decreased in response to OCIF. Urinary excretion of Ca, Pi, and creatinine did not change significantly after injection of OCIF or vehicle. In hypercalcemic, tumor-bearing nude mice, a single intraperitoneal injection of OCIF at 20 mg/kg resulted in a dramatic decrease in serum Ca (maximal decrease 2.8 +/- 0.37 mg/dL, n = 11), which continued for up to 24 h. The results suggest that OCIF decreased serum Ca through its inhibitory effect on bone resorption. Furthermore, it is suggested that OCIF has therapeutic potential for the treatment of hypercalcemic conditions such as malignancy-associated hypercalcemia. PMID- 9855458 TI - Study of immunoelectron microscopic localization of cathepsin K in osteoclasts and other bone cells in the mouse femur. AB - The localization of cathepsin K protein in mouse osteoclasts was examined by immunolight and immunoelectron microscopy using the avidin-biotin-peroxidase complex method with anti-cathepsin K (mouse) antibody. With light microscopy, a strong immunoreaction for cathepsin K was found extracellularly along the bone and cartilage resorption lacunae and detected intracellularly in vesicles, granules, and vacuoles throughout the cytoplasm of multinuclear osteoclasts and chondroclasts attached to the surface of the bone or cartilage. Mononuclear cells, probably preosteoclasts, some distance from the bone also contained a few cathepsin K-positive vesicles and granules. Cathepsin K was sometimes found in the cisternal spaces of the rough endoplasmic reticulum and vesicles of the Golgi apparatus with electron microscopy of the basolateral region of the osteoclasts. Cathepsin K-positive vesicles and granules as lysosomal compartments were present in various stages of fusion with vacuoles as endosomal compartments that contained fragmented cathepsin K-negative fibril-like structures. Some of the vacuoles (endolysosomes), which seemed to be formed by this process of fusion, contained cathepsin K-positive vesicles and fibril-like structures that did not show the regular cross striation of type I collagen fibrils. In the apical region of the osteoclasts, cathepsin K-positive vesicles and pits had already fused with or were in the process of fusing with the ampullar extracellular spaces. There were large deposits of cathepsin K on fragmented fibril-like structures without regular cross striation in the extracellular spaces, as well as on and between the cytoplasmic processes of the ruffled border. There were also extensive deposits of cathepsin K on the type I collagen fibrils with cross striation in the bone resorption lacunae. Osteoblasts and osteocytes were negative for cathepsin K. In the immunocytochemical controls, no immunoreaction was found in the osteoclasts or preosteoclasts, or on the collagen fibrils in the resorption lacunae. The results indicate that cathepsin K is produced in mature osteoclasts attached to the bone and secreted into the bone resorption lacunae. The findings suggest that cathepsin K participates in the extracellular degradation of collagen fibrils in the resorption lacunae and in the subsequent degradation of the fragmented fibrils in the endolysosomes. It is also suggested that cathepsin K degrades the organic cartilage matrix. PMID- 9855459 TI - Matrix mineralization in MC3T3-E1 cell cultures initiated by beta glycerophosphate pulse. AB - MC3T3-E1 cells, grown in the presence of serum and ascorbate, express alkaline phosphatase and produce an extensive collagenous extracellular matrix that can be mineralized by the addition of beta-glycerophosphate (beta-GP). In the present work, we study the influence of concentration and duration of beta-GP treatment on the mineralization pattern in 4-week-old cell cultures. Amount and structure of mineral deposition were monitored by von Kossa staining, light, and electron microscopy, as well as small-angle X-ray scattering (SAXS) of unstained specimens. SAXS measures the total surface of the mineral phase and is therefore preferentially sensitive to very small crystals (typically <50 nm). It was used to determine the ratio (M) of small crystals to collagen matrix. A variety of mineralization patterns was observed to occur simultaneously, some associated with collagen within nodules or in deeper layers of the cultures and some independent of it. At a beta-GP concentration of 10 mmol, mineralization was initiated after about 24 h and continued to increase, irrespective of whether the high level of beta-GP was maintained or reduced to 2 mmol. With shorter pulses (<24 h), no significant mineralization was observed in the week following beta-GP pulse. With continuous treatment at 5 mmol beta-GP, the first signs of mineralization were detected 14 days after the beginning of treatment in the 4 week-old cultures, but no mineralization at all occurred at lower beta-GP concentrations. When cells were grown without ascorbic acid for 4 weeks, only two cell layers without collagen matrix were found. In these cultures, no mineralization detectable by SAXS could be induced with beta-GP. These data indicate that, in viable cells, high doses of beta-GP are essential for the nucleation of mineral crystals, but not for the progression of mineralization once crystals had been nucleated. In contrast, when 4-week-old cell cultures were devitalized, M was found to increase immediately, even at 2 mmol beta-GP. These results suggest that, in MC3T3-E1 cell cultures, cell viability is essential for prevention of spontaneous mineralization of the extracellular matrix. PMID- 9855460 TI - Effects of vitamin D3, 17beta-estradiol, vasoactive intestinal peptide, and glutamate on electric coupling between rat osteoblast-like cells in vitro. AB - Osteoblast-like cells express receptors for various hormones and neurotransmitters that induce widespread actions in the bone to which intercellular communication and its modulation may contribute. Therefore, we examined the effects of the osteotropic hormones vitamin D3 (vitD3) and 17beta estradiol (17beta-E2) as well as the neurotransmitter vasoactive intestinal peptide (VIP) and the excitatory amino acid glutamate (Glu) on gap junctions between rat osteoblast-like (ROB) cells in vitro. Electric coupling was measured by simultaneous intracellular recordings from neighboring cells. The coupling factor (cf) was calculated from membrane potential changes induced by alternate current injections into both cells. In ROB cells cf was increased by 5 x 10(-8) mol/L vitD3 to 130 +/- 13% (mean +/- SD; n = 6) of the initial value within 5-20 min. This effect was not reversible after washing with control saline for 10-15 min. In six cell pairs, cf was not affected by vitD3 (94 +/- 5%). In three cell pairs superfusion of 10(-8) mol/L E2 reduced cf to 80 +/- 6% within 10 min, whereas, in two cell pairs, this hormone improved cf to 140% within 20 min. Exposure of VIP (3 x 10(-8) mol/L) did not alter cf in the majority of cells (99 +/- 3%; n = 11). In five cell pairs, cf was improved within 5-15 min to 133 +/- 12%, whereas, in one cell pair, cf was reduced to 22% by VIP. In contrast, brief application of Glu (5 x 10(-3) mol/L) decreased cf to 75 +/- 5% (n = 5), whereas, in nine other cell pairs, cf was not affected (96 +/- 5%). The findings indicate that cell-cell coupling of gap junctions between bone cells can be altered by actions of hormones and transmitters in a cell-pair-specific way, which may depend on their functional state. PMID- 9855461 TI - Histomorphometric, physical, and mechanical effects of spaceflight and insulin like growth factor-I on rat long bones. AB - Previous experiments have shown that skeletal unloading resulting from exposure to microgravity induces osteopenia in rats. In maturing rats, this is primarily a function of reduced formation, rather than increased resorption. Insulin-like growth factor-I (IGF-I) stimulates bone formation by increasing collagen synthesis by osteoblasts. The ability of IGF-I to prevent osteopenia otherwise caused by spaceflight was investigated in 12 rats flown for 10 days aboard the Space Shuttle, STS-77. The effect IGF-I had on cortical bone metabolism was generally anabolic. For example, humerus periosteal bone formation increased a significant 37.6% for the spaceflight animals treated with IGF-I, whereas the ground controls increased 24.7%. This increase in humeral bone formation at the periosteum is a result of an increased percent mineralizing perimeter (%Min.Pm), rather than mineral apposition rate (MAR), for both spaceflight and ground control rats. However, IGF-I did inhibit humerus endocortical bone formation in both the spaceflight and ground control rats (38.1% and 39.2%, respectively) by limiting MAR. This effect was verified in a separate ground-based study. Similar histomorphometric results for spaceflight and ground control rats suggest that IGF-I effects occur during normal weight bearing and during spaceflight. Microhardness measurements of the newly formed bone indicate that the quality of the bone formed during IGF-I treatment or spaceflight was not adversely altered. Spaceflight did not consistently change the structural (force-deflection) properties of the femur or humerus when tested in three-point bending. IGF-I significantly increased femoral maximum and fracture strength. PMID- 9855462 TI - Specific bone-protective effects of metabolites/derivatives of tamoxifen and clomiphene in ovariectomized rats. AB - In the ovariectomized (ovx) rat, the nonsteroidal antiestrogens, clomiphene (CLO) and tamoxifen (TAM), at dose levels that prevent development of osteopenia to a degree approaching that of 17beta-estradiol are, in contrast to 17beta-estradiol, only weakly uterotrophic. Metabolites of CLO and TAM might contribute differentially to these effects. Thus, we have evaluated bone protective and uterine effects in ovx rats of two such metabolites: 4-hydroxy CLO, produced by p hydroxylation of CLO; and 4HTA, produced from TAM by stepwise replacement of its dimethylaminoethyl side chain with an acetic acid moiety, accompanied by p hydroxylation. Also reported are effects of D4HTA, the dihydrodesethyl derivative of 4HTA previously characterized as a full estrogen mimetic in vitro. Administration of 4-hydroxy CLO (2.5 mg/kg subcutaneously) 5 days/week for 5 weeks to 3-month-old ovx rats resulted in complete prevention of bone loss and suppression of bone turnover to levels comparable to those of intact controls and to those of ovx animals similarly receiving 17beta-estradiol (10 microg/kg). However, uterine weight in animals receiving 4-hydroxy CLO was 64% less than that in 17beta-estradiol-treated animals. Although 4HTA (3.7 mg/kg s.c.) had a modest uterotrophic effect, it did not prevent bone loss associated with ovariectomy. In contrast, D4HTA (3.6 mg/kg s.c.) partially reduced bone turnover indicators and cancellous bone loss in a manner similar in many ways to that observed in TAM treated ovx animals, but it had no uterotrophic effect. These results suggest that, although 4HTA does not contribute to the bone-protective effect of TAM, 4 hydroxy CLO might augment that of CLO. PMID- 9855463 TI - Paget's disease of bone: benefits of neridonate as a first treatment and in cases of relapse after clodronate. AB - This study assessed the efficacy of 200 mg of aminohexane bisphosphonate (neridronate) administered by intravenous infusion in a single dose or in two separate doses on consecutive days in 32 patients (16 males and 16 females, average age 66 years) affected by active Paget's disease of bone. Fifteen patients had never been treated with any antiresorptive agent and 17 had had unsatisfactory results from a prior clodronate treatment. All of the latter patients had failed to enter a remission stage (i.e., normalization of bone turnover was not reported at any time during treatment) and had had a full relapse within 6 months after clodronate infusion. In the present study bone specific alkaline phosphatase (bAp), deoxypyridinoline (dPyr), and N- and C terminal polypeptide of collagen type 1 (Ntx, Ctx) were determined before neridronate administration and at 1, 3, 6, and 12 months thereafter. Basal values of bAp were 51.7 +/- 2.3 microg/L, range 31.7-92.5 (normal range 6.2-23.6). No statistical differences in markers of bone turnover were evident in the basal state between new pagetic patients (bAp = 55.1 +/- 4.1) and those suffering a relapse after clodronate (bAp = 48.8 +/- 2.6). Neridronate induced an average percent change from baseline in excess bAp of 68.0 +/- 4.3 and in excess dPyr, Ntx, and Ctx of 68.1 +/- 11, 60.6 +/- 8.5, and 86.7 +/- 7.8, respectively. Markers of bone resorption declined more slowly in patients treated previously with clodronate, although the average change in percent decrement from baseline in excess bAp as well as in excess of bone resorption markers was not different from that registered in untreated pagetic patients. Response to treatment, defined as a percent decrement from baseline in excess bAp of 50% or more at any time during the 12-month follow-up, was observed in 27 patients (84.4%). Remission (a drop in bAp to within normal range) was achieved in 21 patients (65.6%) and was maintained in 12 at 12-month follow-up, with no significant differences between either 1- or 2-day infusions, or between new pagetic patients and those relapsing after clodronate. In 15 of 21 patients requiring analgesics to alleviate bone pain, pain was reduced or completely alleviated in 8. A slight, short-lived acute phase reaction (fever and/or arthromyalgia) occurred in 6 patients. To summarize, 200 mg of intravenous neridronate, in one or two doses, significantly reduced the biochemical indices of disease activity in the majority of patients, showing a normalization of bAp in more than 60%. We conclude that neridronate can be used safely in the treatment of patients with Paget's disease of bone either as a first bisphosphonate treatment or as retreatment for patients relapsing after clodronate. PMID- 9855464 TI - Tumor expression studies indicate that HEM-1 is unlikely to be the active factor in oncogenic osteomalacia. AB - HEM-1 was isolated as a putative factor responsible for oncogenic osteomalacia by Kumar et al. (Proc Assoc Am Phys 107:296-305; 1995). The cDNA was identified on the basis of PTH-like immunoreactivity; however, no studies have been reported of the expression of HEM-1 mRNA in oncogenic osteomalacia tumors. In this study, expression of HEM-1 mRNA was investigated in two oncogenic osteomalacia tumors and in a series of normal tissues. An HEM-1 PCR product was amplified from a cDNA library from one of the tumors, with six base changes identified, as compared with the published sequence. No expression was detected, however, in the oncogenic osteomalacia tumors either by Northern blot analysis or by reverse transcriptase PCR. This indicates that, although a region of HEM-1 sequence is present in the tumor cell cDNA library, any HEM-1 expression must be at very low levels. It is unlikely, therefore, that the HEM-1 product is the active factor responsible for oncogenic osteomalacia. In the normal tissues examined, human placenta, fibroblasts, parathyroid gland, liver, fetal bone, and rat kidney cortex, HEM-1 mRNA was not detected, suggesting that it does not have a physiological role in these tissues. PMID- 9855465 TI - High prevalence of vitamin D deficiency and reduced bone mass in elderly women with Alzheimer's disease. AB - Patients with Alzheimer's disease (AD) are at increased risk for falls and hip fractures. To better understand causes and prevention, we measured bone mineral density (BMD) in the second metacarpals of 46 ambulatory elderly women with AD and analyzed its relation to serum biochemical indices, sunlight exposure, and vitamin D intake. BMD was significantly less than in age-matched controls. In 26% of AD patients, the serum 25-hydroxyvitamin D (25-OHD) concentration was at a deficient level (5-10 ng/mL), and in 54% it was at an osteomalacic level (<5 ng/mL). Concentrations of ionized calcium were significantly lower in patients. Conversely, concentrations of serum bone Gla-protein and urinary hydroxyproline in patients were significantly higher than in controls. BMD correlated positively with 25-OHD concentration (p = 0.0041) and negatively with parathyroid hormone (PTH) concentration (p = 0.0022). PTH was higher in patients than in controls, and correlated negatively with 25-OHD (p < 0.0001). Many AD patients were sunlight-deprived and consumed less than 100 IU of vitamin D per day. We concluded that vitamin D deficiency due to sunlight deprivation and malnutrition, together with compensatory hyperparathyroidism, contributes significantly to reduced BMD in AD patients. Low BMD increases risk of hip fractures in patients with AD, but may be improved by vitamin D supplementation. PMID- 9855466 TI - Taste in chimpanzees. III: Labeled-line coding in sweet taste. AB - In peripheral taste the coding mechanism remains an enigma. Among coding theories the "across-fiber pattern" argues that activity across fibers codes for taste, whereas the "labeled line" claims that activity in a particular set of fibers underlies a taste quality. We showed previously that chimpanzee chorda tympani taste fibers grouped according to human taste qualities into an S-cluster, responding predominantly to sweet stimuli, a Q-cluster, sensitive to bitter tastants, and an N-cluster, stimulated by salts. The analysis showed that information in the S-line suffices to distinguish stimuli of one taste quality from the others. However, one condition for the labeled line remained: that blockage of activity in a particular line must cause blockage of one taste quality, but of no other, or its onset give rise to the sensation of a taste quality. Here we studied this requirement with gymnemic acids and miraculin. In humans and chimpanzees, gymnemic acids suppress the sweet taste of all sweeteners whereas miraculin adds a sweet taste quality to sour stimuli. Gymnemic acids also abolish miraculin-induced sweet taste. We found that gymnemic acids practically abolished the response to every sweetener in the chimpanzee S-cluster. Equally important, they had no effect on the responses of the Q- and N-clusters. After miraculin, the S-cluster fibers responded to acids as well as to sweeteners, although they had not responded to acids before miraculin. Gymnemic acids abolished this miraculin-induced response to acids and responses to sweeteners in the S-fibers. These results link the sweet taste quality to activity in fibers of the S-cluster. Thus the S-cluster fibers satisfy the definition of the labeled line theory: "that activity in a particular fiber type represents a specific taste quality." PMID- 9855467 TI - Defensive behaviors in wild and laboratory (Swiss) mice: the mouse defense test battery. AB - The development of laboratory rodent models for elicitation and measurement of a range of defensive behaviors raises the question of the relationship between defense in these animals and those of their wild congeners. To evaluate this relationship for mice, defensive responses to an anesthetized rat were compared for fourth-generation laboratory-bred wild mice and Swiss CD-1 (Swiss-Webster derived) laboratory mice in a Mouse Defense Test Battery. Wild mice showed enhanced levels of both freezing and flight, fleeing from distant approach of the predator in several situations and fleeing more quickly than the Swiss mice. However, Swiss mice did flee upon contact with the rat and also showed levels of several other defensive behaviors (risk assessment, defensive threat, and attack) that were often reliably higher than those of the wild mice. However, when wild mice were prevented from fleeing, their levels of defensive threat and attack were as high as, or at very short prey-predator distances higher than, those of the Swiss mice. These findings suggest that flight and freezing are the major defensive behaviors reduced in Swiss mice and that these reductions allow the appearance of higher levels of additional defensive behaviors in the laboratory animals. However, although Swiss mice do show lower levels of flight and freezing, their patterns of defensive behavior are sufficiently similar to those of wild mice that they provide adequate subjects for research on the biologic bases of defensive behavior. A final experiment indicated that when wild mice are familiarized with a chamber providing a place of concealment, they flee directly to this chamber on presentation of a rat, indicating that flight is a targeted response and not simply an abrupt increase in forward locomotion. Over 10 rat presentation trials with a blocked chamber entrance, however, this response declines. PMID- 9855468 TI - Selection of macronutrients by goldfish operating self-feeders. AB - In the wild, both food availability and an animal's requirements may fluctuate. Given the choice to feed on different diets, animals may exhibit preferences in their selection. Although macronutrient selection has been investigated in mammals and birds, few attempts have been made to investigate dietary selection in fish. In the present study we investigate the feeding of goldfish on three "pure" macronutrient diets. Twelve single goldfish and one group of four goldfish, initially weighing 163.1 +/- 7.4 and 70.2 +/- 8.0 g, respectively, were maintained in 54-liter tanks at 20 degrees C under a 12 h:12 h light-dark (LD) cycle. Three pelleted diets were made containing 50% of each macronutrient: protein (casein + gelatin), fat (fish oil + soy oil), or carbohydrate (dextrin). All the diets included mixtures of vitamins and minerals, a binder (sodium alginate), and cellulose as a filler. During trials, three self-feeding devices delivering the protein, fat, or carbohydrate diet were installed in each aquarium. Goldfish soon learned to select from among the three diets. Overall daily food and digestible energy demands differed significantly between macronutrients. The demand for protein was lower than that for carbohydrate (16.6 and 33.7 g/kg BW/day, respectively), and the digestible energy demand for protein was lower than for fat (108.6 and 308.5 kJ/kg BW/day, respectively). Goldfish made their dietary selections on the basis of energy content, so that food demand increased to compensate for changes in the digestible energy density of the diets. Correlation analysis revealed a significant positive relationship between carbohydrate and fat demand (n = 8), whereas there were significant negative (n = 3) and positive (n = 1) relationships between protein and fat. Different daily patterns for the selection of each macronutrient were detected in some fish (i.e., carbohydrate during daytime, protein at the beginning of the night, and fat during the transition phase). In short, our results demonstrated the ability of goldfish to compose their own diet from three basic macronutrients, highlighting the complex pattern of behavior in their selection. Therefore, these new findings should be considered when studying feeding rhythms and the design of diets for fish. PMID- 9855469 TI - Effects of melatoninergic agonists on light-suppressed circadian rhythms in rats. AB - This study addressed the question whether light-suppressed circadian rhythms in cardiovascular parameters in rats could be restored by melatonin and a synthetic analogue. Blood pressure, heart rate, and locomotor activity were monitored by radiotelemetry in six Sprague-Dawley rats. After synchronization to a 12:12 light/dark (LD) schedule (lights on at 0700 hours, 100 lux), rats were kept in constant light (LL) of low intensity (5-10 lux) for 11 weeks. After 3 weeks of LL, rats received daily intraperitoneal (i.p.) injections at 1900 hours of vehicle, the melatonin agonist S-21767 (5 mg/kg) and melatonin (1 mg/kg). Spectral power, 24-h amplitudes and the differences between day and night means were calculated as measures of circadian rhythmicity. During LL a lengthening of the endogenous period to 26 h was observed, which was accompanied by a continuous decrease in circadian amplitude in all parameters monitored until, in the third week of LL, circadian rhythmicity was almost abolished. Neither vehicle, S-21767 nor melatonin were able to restore circadian rhythms in blood pressure and locomotor activity. In contrast, both agonists induced circadian rhythmicity in heart rate in two out of six rats. The day/night difference in heart rate of all animals was significantly increased by S-21767 and, to a smaller extent, by melatonin, whereas the circadian amplitude was not affected. In conclusion, melatonin and the synthetic agonist were able to partially synchronize circadian rhythmicity in heart rate during constant light, but could not restore circadian rhythms in blood pressure. PMID- 9855470 TI - Forced dissociation of food- and light- entrainable circadian rhythms of rats in a skeleton photoperiod. AB - To investigate the control over drinking and feeding behavior by the light entrainable circadian pacemaker, rats were maintained in a 12 h:12 h skeleton photoperiod for 36 days with both food and water restricted to the subjective day. During the restriction period most of the food and water intake was concentrated during the first 4 h of the subjective day. The subsequent release into ad lib conditions under the skeleton photoperiod or continuous dark showed that this increased ingestion during the subjective day persisted for up to 6-10 days. This may indicate the entrainment of a food-entrainable oscillator, which has hitherto been investigated solely in anticipatory activity. The daily activity rhythm returned to its original phase position. Thus the phase of the overt daily rhythm is eventually only determined by the phase of the light entrainable oscillator (LEO) but can temporarily be influenced by the restriction of food and water access. This restriction apparently stimulated a food entrainable oscillator in antiphase with the light-entrainable oscillator but failed to phase shift or entrain the light-entrainable oscillator. PMID- 9855471 TI - Effects of fadrozole on sexual displays and reproductive activity in the female canary. AB - We used fadrozole, a potent inhibitor of the aromatization of androgens to estrogens, to investigate the influence of estradiol on copulation solicitation displays (CSD) and reproductive activity in female canaries (Serinus canaria). Systemic injections of fadrozole during 10 consecutive days were effective in reducing plasma estradiol concentrations in adult female canaries submitted to photostimulation. Fadrozole provides a powerful tool for limiting an individual's exposure to estradiol, and the results of this study emphasize the influence of estradiol secretion in the regulation of behavioral transitions along the reproductive cycle of the female canary. When females were injected at the beginning of photostimulation, the emergence of copulation solicitation displays in response to conspecific songs was delayed. When females were injected later, after they were sexually active, the fadrozole treatment did not affect sexual displays. These results are consistent with the hypothesis that a threshold level of estradiol is critical to activate the neural circuitry mediating the copulation solicitation displays response in the female canary. They also suggest that the magnitude of sexual response is not related in a dose-dependent manner to estrogen concentrations observed during the period of sexual responsiveness. When females were injected at the beginning of photostimulation, egg-laying was delayed; when females were injected later, after they were sexually active, the fadrozole treatment dramatically reduced egg-laying and prevented incubation. PMID- 9855472 TI - Adolescent acceptance of different foods by obesity status and by sex. AB - This work investigated possible differences in food preference between obese and normal-weight adolescents, as well as between female and male seventh-grade pupils (average age = 13.1 years). Survey of affinities for nine food groups was determined with a facial hedonic scale. The children were divided according to their BMI into normal-weight and obese group, respectively. In this sample of adolescents, we could not find any proof for the theory of increased preference for fattening foods by obese compared to normal-weight people. In fact, our normal-weight group showed greater liking of sweets, meat, and cereals than the obese group. Normal-weight boys also preferred meat and sweets more than normal weight girls. We consider psychological and social factors (e.g., perception of ideal body shape or social stigmatism of obesity) as the most logical explanations for the observed differences. Further research should clarify whether food preference plays a significant role in the etiology of adolescent obesity. PMID- 9855473 TI - Modulation of mice anxiety in response to cat odor as a consequence of predators diet. AB - The effectiveness of predator odours as repellents was assessed, and the behavioral antipredatory responses were characterized. Mice had free access to an unfamiliar runway containing different olfactory stimuli: modelling clay, or feces of a cat subjected either to a vegetarian or a carnivorous diet. The first experiment revealed various indices of a spontaneous behavioral pattern that included exploratory activity, different kinds of emotionality, and a range of active or passive defensive reactions until the appearance of absence of risk assessment strictly related to presence or absence of anxiety. These reactions differ with larger responses to feces resulting from a carnivorous as opposed to vegetarian diets. In the second experiment, chlordiazepoxide (0, 2.5, 5, or 7.5 mg/kg) had a dose-related anxiolytic effect on exploration in mice of both vegetarian and carnivorous groups but could not totally reverse the strong anxiogenic effect of carnivorous stimulus on defensive mechanisms. These differences are related to the nature of the mammalian cues. This paradigm may be a fear-motivated model of animal anxiety. PMID- 9855474 TI - Behavioral thermoregulatory responses of single- and group-housed mice. AB - The ambient temperature (Ta) to house and study laboratory rodents is critical for nearly all biomedical studies. The ideal Ta for housing rodents and other animals should be based on their thermoregulatory requirements. However, fundamental information on the behavioral thermoregulatory responses of single- and group-housed rodents is meager. To address this issue, thermoregulatory behavior was assessed in individual and groups of CD-1 mice housed in a temperature gradient. Mice were housed in groups of five or individually while selected Ta and motor activity were monitored. Single- and group-housed mice displayed a circadian oscillation of selected Ta and motor activity with relatively warm T(a)s of approximately 29 degrees C selected during the light phase; during the dark phase selected Ta was reduced by 4 degrees C, whereas motor activity increased. Selected Ta of aged (11 months old) mice housed individually was approximately 1.0 degrees C warmer than the group-housed mice. Thermal preference of younger mice (2 months old) was similar for single- and group-housed animals. The operative Ta of mice housed in standard facilities was estimated by measuring the cooling rate of "phantom" mice modeled from aluminum cylinders. The results show that the typical housing conditions for single- and group-housed mice are cooler than their Ta for ideal thermal comfort. PMID- 9855475 TI - Ultra-wideband electromagnetic pulses and morphine-induced changes in nociception and activity in mice. AB - Mice were exposed to ultra-wideband (UWB) electromagnetic pulses averaging 99-105 kV/m peak amplitude, 0.97-1.03 ns duration, and 155-174 ps rise time, after intraperitoneal administration of saline or morphine sulfate. They were then tested for thermal nociception on a 50 degrees C surface and for spontaneous locomotor activity and its time profile over 5 min. Analysis of results showed no effect of UWB exposure on nociception and activity measures in CF-1 mice after 15 , 30-, or 45-min exposure to pulses at 600/s or after 30-min exposure to UWB pulses at 60/s. Similarly, no effect was seen in C57BL/6 mice after 30-min exposure to pulses at 60/s or 600/s. Although trends in morphine-modified measures seen with UWB pulse repetition frequency could be expected because of increased levels of low-frequency energy, no significant change was seen in normal or morphine-modified nociception or activity after UWB exposure. This indicated lack of effect of the UWB pulses used in these experiments on nervous system components, including endogenous opioids, involved in these behaviors. PMID- 9855476 TI - Brain insulin response to feeding in the rat is both macronutrient and area specific. AB - Using microdialysis, we showed recently that hypothalamic immunoreactive insulin (IRI) levels increased after a meal of chow and decreased in response to a fat meal. In the present study, we have compared extracellular hypothalamic and extrahypothalamic basal IRI levels and investigated the effect of meals composed exclusively of either carbohydrates (85% starch, 15% sucrose) or casein on both plasma and medial hypothalamic (PVN-VMH) insulin. The response of IRI to a carbohydrate meal was also investigated in the cerebellum. Basal hypothalamic IRI was twofold higher in the hypothalamus as compared to the cerebellum (33 +/- 4 and 15 +/- 2 pg/mL, respectively). Hypothalamic IRI increased twofold in response to the carbohydrate meal (72 +/- 15 pg/mL) but remained unchanged during the casein meal. No IRI change was found in the cerebellum after a meal of carbohydrates (16 +/- 2 pg/mL). Insulinemia was increased by both the carbohydrate and the casein meal. However, the protein-induced increase was less pronounced (maximum + 359% compared to 1650% for carbohydrates). The present data show a dual specificity of brain insulin response to feeding; in addition to the macronutrient specific variations, a regional specificity was also observed. Taken together with previous observations, the present data are in favor of an involvement of PVN-VMH insulin in the control of feeding and macronutrient specific appetites. PMID- 9855477 TI - Glucose, but not fat, phase shifts the feeding-entrained circadian clock. AB - To study the ability of single macronutrients to entrain or phase shift the feeding entrainable circadian oscillator, rats with lesions of the suprachiasmatic nucleus were first maintained on a single daily meal of lab chow until robust anticipatory approaches to the feeder or anticipatory wheel running was established. The meal time was then delayed by 8 h and chow was replaced with a 25-mL solution of 0.2% saccharin or 25 mL of saccharin plus 15 g of glucose. For other phase shifts, rats received either 6 mL of vegetable oil or mineral oil for 2 consecutive days. Consumption of about 6 g (24 kcal) or more of glucose resulted in robust delaying transients on the days after ingestion, whereas saccharin induced only small delays consistent with the initiation of a free running rhythm with a period greater than 24 h. Surprisingly, consumption of 5.5 g of vegetable oil (47 kcal) did not result in delays greater than those in rats receiving mineral oil. The introduction of oil also produced a severe reduction in approaches to the feeder which could be alleviated by placing inaccessible chow in the feeders between oil meals. Phase shifts with oil were repeated with rats housed in wheels using anticipatory wheel running as a phase marker to assess whether the lack of phase shifts with fat was apparatus dependent. As was the case with approach behavior, anticipatory wheel running was not significantly delayed by vegetable oil consumption. These results indicate that a simple monosaccharide, glucose, has zeitgeber properties for the feeding entrainable oscillator. Vegetable oil, despite a higher caloric content, may be ineffective because of slower gastric emptying and nutrient absorption or because fat is not a good zeitgeber for the feeding entrained circadian oscillator. PMID- 9855479 TI - Seasonality of meal patterns and hormonal correlates in red deer. AB - Two groups of six adult, castrated, male red deer were housed under natural daylength conditions and at ambient temperature at 57 degrees N and fed ad lib. (AL) or at a fixed rate of 50 g/kg0.75 initial liveweight per day throughout the study (restricted, R). Mean daily intakes of AL animals were higher during periods of long daylength than during short daylength (p < 0.001). The higher rates of food intake during periods of long daylength were a function of greater meal durations (p < 0.001), shorter inter-meal intervals (p < 0.001) and higher (p < 0.001) mean rates of ingestion (g/min). In both groups mean plasma concentrations of prolactin, T3, T4, and insulin-like growth factor 1 (IGF-1) were higher (p < 0.001) during long daylength than short daylength although changes in thyroid hormone profiles were much less marked in AL animals. Insulin and growth hormone (GH) profiles exhibited no consistent seasonal trend. Mean plasma concentrations of T3 were higher in AL than in R animals. Mean plasma IGF 1 concentrations during long days were consistently greater in the AL than R animals. It is concluded that the effects of seasonal changes in daylength on appetite and food intake are expressed through changes in both the duration of daylight periods per se and in underlying seasonal changes in physiology and associated meal patterns and eating rates. It is concluded that the roles of T3, IGF-1, and prolactin in the expression of seasonal changes in appetite should be investigated further and, particularly, their effects on other hormone profiles and liver and gut function. PMID- 9855478 TI - Soybean- and olive-oils-enriched diets increase insulin secretion to glucose stimulus in isolated pancreatic rat islets. AB - Islets isolated from rats fed a lipid-enriched diet have shown an impairment of insulin secretion, but there is no available data comparing the effect of diet containing different dietary fat. This may be important in preventing or facilitating the establishment of diabetes. In this study, the effect of diets enriched (10%) with different fatty acids on insulin secretion by isolated pancreatic islets was investigated. The sources of the fatty acids tested were: saturated long chain from animal fat (AF), polyunsaturated from soybean oil (SO), and monounsaturated from olive oil (OL). The results were compared with those from rats receiving a diet enriched (10%) with a balanced mixture of fatty acids (the same proportion of AF, SO, and OL). The effect of fat-rich diets on insulin release was tested in vivo by giving a glucose load (glucose tolerance test-GTT) and in vitro in perfused islets. The mechanism involved was also examined by measuring 45Ca2+ and 86Rb+ fluxes, GLUT-2 content, and glucose oxidation in isolated islets. A significant increase of insulin secretion and glucose oxidation without any alteration of the ionic movements were detected in islets from SO and OL rats. GLUT-2 content was increased in islets of the OL group but diminished in AF rats. The results led us to postulate that soybean and olive oils may increase the response of insulin secretion to glucose stimulus in pancreatic islets. PMID- 9855480 TI - Untangling the effects of hunger, anxiety, and nausea on energy intake during intravenous cholecystokinin octapeptide (CCK-8) infusion. AB - Infusions of cholecystokinin (CCK) may exert their effects on appetite by inducing feelings of nausea or anxiety. In this double blind, placebo controlled crossover study, the impact of these effects on appetite were examined. Fifteen male subjects received a 20 min i.v. infusion of cholecystokinin octapeptide (CCK 8) (4 ng/kg/min) or saline. The infusion commenced 20 min after a soup preload and 10 min before an ad libitum test meal. Visual analogue scales of appetite and mood were measured over 3 h, and subjects were instructed to report any other sensations they experienced over this time. CCK-8 significantly reduced premeal hunger, elevated premeal anxiety, and reduced energy intake at the ad libitum test meal. Meal duration and rate of eating (kcal/min) were also significantly reduced after CCK-8. After the smaller meal with CCK-8, hunger rose quickly to a higher level than with placebo. The return of hunger was commensurate with the smaller amount of energy consumed and indicated that CCK did not exert an enduring effect on hunger suppression. A significant correlation was found between the reduction in energy intake and hunger (r = 0.75 p < 0.01), but not with anxiety (r = 0.15 not significant). Analyses were performed separately on subjects who did (n = 8), or did not (n = 7) report gastrointestinal disturbance. Energy intake was reduced by 56.6% and 44.6%, respectively. These results indicate that, although feelings of anxiety and nausea may accompany CCK infusions, they are not necessary for the effects of CCK on appetite. These data provide support for a role of CCK in satiety. PMID- 9855481 TI - The influence of odor type on the discrimination and identification of odorants in multicomponent odor mixtures. AB - Using a limited set of odorants, previous studies have indicated that the ability of humans to discriminate and identify the components of olfactory mixtures is limited to approximately four. However, the ability to generalize these results may have been limited by specific neural or cognitive interactions among the particular odorants used. In the present experiment, 41 subjects examined the influence of odor type (different individual odorants), from two very different odor sets, on the perception of the components of complex mixtures. One set contained odors that were selected by an expert panel to blend well in mixtures (good blenders), whereas the other contained odors that blended poorly in mixtures (poor blenders). The stimuli were common, dissimilar odorants of equivalent, moderate intensity, each of which was a single chemical. A computer controlled air dilution olfactometer delivered a single odorant or a mixture containing up to eight odorants. Although the poor blenders were more easily discriminated, this superiority was displayed within a narrow range, and the ability of subjects to identify mixture components with either odor set was limited to approximately four. The results indicate that, whereas odor type can alter which odorants will be perceived in a mixture, the limited capacity to discriminate mixture components is independent of the type of odorants. These findings are discussed in terms of their implications for olfactory coding. PMID- 9855482 TI - Central lead administration induces natriuretic and kaliuretic effects in rats. AB - The aim of the present experiments was to discern whether central acute lead injections affect brain control of renal function. Adult Wistar male rats received third-ventricle injections of lead acetate in three different doses (0.03, 0.3, and 3.0 nmol/rat). Lead acetate induced a significant increase in renal excretion of sodium and potassium. Pretreatment with losartan, a selective angiotensin II AT1 receptor antagonist (10.8 nmol/rat into the third ventricle 10 min before central lead injection), inhibits lead-induced natriuretic and kaliuretic effects. In addition, pretreatment with gadolinium, a calcium-channel blocker (0.3 nmol/rat into the third ventricle 20 min before central lead administration), reversed the increase in renal excretion of sodium and potassium provoked by central lead administration. Taken together, the data presented here suggest that lead injected into the third ventricle increases renal excretion of sodium and potassium by a mechanism that depends on the functional integrity of central angiotensin II AT1 receptors and calcium channels. PMID- 9855483 TI - Production of serotonin syndrome by 8-OH DPAT in Cryptotis parva. AB - We have previously shown that the 5-HT2A/C agonist, DOI, potently and in a dose dependent manner produces the head-twitch response in the least shrew (Cryptotis parva) via the activation of serotonergic 5-HT2A receptors. The purpose of the present study was to determine whether activation of 5-HT1A receptors by its selective agonist, 8-OH DPAT, can induce the serotonin syndrome (SS) in this species. In the rat, the symptoms of SS include: forepaw splaying, hindleg abduction, forepaw treading, flat body posture, tremor, and straub tail. Intraperitoneal (i.p.) administration of 8-OH DPAT produced four classic symptoms (forepaw splaying, hindleg abduction, forepaw treading, and straub tail) of SS in the least shrew in a dose-dependent manner in the 30-min observation period. The mean total cumulative score for all components of SS also significantly increased in intensity in a dose-dependent fashion. Administration of selective 5-HT1A antagonists [S(-)UH 301 or NAN-190] potently blocked the 8-OH DPAT-induced mean total SS score in a dose-dependent manner. Moreover, these antagonists had similar potencies as indicated by their identical ID50 values (0.5 and 0.52 mg/kg respectively). However, unexpectedly and unlike the published findings in the rat, the nonselective 5-HT1A antagonist with b-blocking activity, propranolol, failed to attenuate the induced response in this species. As was expected, the selective 5-HT2A/C antagonist, SR 46349B, did not affect the intensity 8-OH DPAT induced symptoms. Overall, these data suggest that the SS produced by 8-OH DPAT in the least shrew is mediated via the activation of serotonergic 5-HT1A receptors. In addition, propranolol is not a useful 5-HT1A antagonist in this species. PMID- 9855484 TI - Responses of lean and obese Zucker rats to centrally administered leptin. AB - Obese (Lepr(fa)/Lepr(fa)) Zucker rats have a missense mutation in the leptin receptor gene. One amino acid substitution in the extracellular domain common to all known leptin receptor proteins results from this mutation. Obese Zucker rats are unable to respond behaviorally to leptin which is peripherally administered. However, conflicting reports exist on whether obese Zucker rats can respond to centrally administered leptin. The purpose of this study was to determine whether obese Zucker rats responded behaviorally and metabolically to intracerebroventricularly (i.c.v.) administered leptin and to compare the responses of lean and obese Zucker rats. We found that both lean and obese Zucker rats had similar body weight and food intake responses when administered a single i.c.v. leptin injection in a range of doses (1.25, 2.5, 5, and 10 microg), as well as daily i.c.v. administered leptin for five consecutive days. Both single and daily leptin administration also decreased respiratory quotient (RQ) similarly in lean and obese Zucker rats, indicating mobilization of fat as an energy source for leptin-treated rats. After withdrawal of daily leptin treatment, lean and obese Zucker rats exhibited different recovery responses. It is concluded that obese Zucker rats can respond to exogenous leptin when leptin is delivered into the brain ventricles. PMID- 9855485 TI - Hyperphagia in pre-fed rats following oral delta9-THC. AB - Using a pre-feed paradigm, the effects of orally-administered delta9 tetrahydrocannabinol (THC) on low baseline levels of nocturnal feeding were assessed. Following 2-h access to a palatable wet mash diet at dark onset, adult male Lister hooded rats (Charles River) were treated with either sesame seed oil vehicle or delta9- tetrahydrocannabinol (0.063, 0.12, 0.25, 0.5, 1.0, or 2.0 mg/kg). One hour later, rats were allowed ad libitum access to standard chow, and intakes were monitored over the subsequent 24 h. Doses of 0.5, 1.0, and 2.0 mg/kg produced substantial hyperphagia during the first hour of testing. Subsequently, rats compensated for their overconsumption so that 24-h intakes were similar in all conditions. The data confirm anecdotal reports of the orexigenic actions of exogenous cannabinoids and suggest a critical role for endogenous cannabinoid systems in the regulation of appetite. PMID- 9855486 TI - Infection with Schistosoma mansoni in mice induces changes in nociception and exploratory behavior. AB - In this study, CD-1 mice were infected percutaneously with 1600 cercariae of Schistosoma mansoni and their pain sensitivity and exploratory behavior were analyzed in well-standardized tests (hot-plate, hole-board, open-field, novel object investigation and black/white box). Schistosome infection produced body weight reduction, increased analgesia, induced changes in the number of fecal pellets emitted during the hole-board and the black/white tests, induced decreased locomotion in the open-field, decreased sniffing, rearing, wall-rearing and time spent in exploratory activity. The infection also lengthened the latency time to the first transition from the white into the black compartment in the black/white box, index of enhanced anxiety. The present findings indicate that the analgesia is one of the main effects of the disease suggesting that schistosome infection induces maladaptive response in exploratory behavior and in locomotor activity of the host associated with altered motivational and attentional levels. Furthermore, though mouse behavioral changes appear to be similar to those observed in parasite/host systems where the changes are supposed to be adaptive for the parasite, in the case of Schistosoma/mouse system, the changes in host behavior resulted to be not adaptive for the parasite. PMID- 9855487 TI - A "pre-satiety sequence" in rats drinking sucrose solutions. AB - Food-deprived rats were given intermittent access or, in a replication, continuous access to a 20% sucrose solution. Both drinking and nondrinking behaviors were recorded. During the ensuing drinking bout, latency of lapping after snout apposition, and duration of lapping bouts, did not change. Approaches to the sipper tube usually eventuated in lapping, though aborted approaches increased in frequency late in the session. Drinking was interrupted by nondrinking behaviors which appeared in a characteristic rostro-caudal sequence: partial head withdrawal from the drinking aperture, then full head withdrawal, then movements of the front paws, then movement of the hind paws with full-body locomotion. All these behavioral changes occurred before there was any appreciable reduction in rate of lapping. PMID- 9855488 TI - Visceral afferent participation in delayed satiation following NMDA receptor blockade. AB - We previously reported that rats increase their food intake, but not water intake, following intraperitoneal (i.p.) injection of MK-801, a noncompetitive antagonist of N-methyl-D-aspartate (NMDA)-activated ion channels. The drug appears to specifically interfere with signals that participate in satiation, thereby prolonging the meal and increasing its size. The mechanism by which delayed satiation occurs is not known. However, some well-studied satiety signals are carried by visceral sensory fibers that innervate the abdominal viscera. We hypothesized that MK-801 might increase food intake by interfering with satiety signals transmitted by visceral afferent neurons. To test this hypothesis, we examined MK-801's effect on food intake in rats systemically treated with capsaicin, a neurotoxin that destroys small unmyelinated visceral afferent neurons. Capsaicin treatment significantly attenuated increased sucrose intake following MK-801. We also investigated whether the effects of MK-801 on food intake would persist in rats treated with total subdiaphragmatic vagotomies. MK 801 increased the intake of 15% sucrose by sham-vagotomized rats, while vagotomized rats did not increase their intake following MK-801. Taken together, these results support the hypothesis that capsaicin-sensitive visceral sensory neurons are involved in increased food intake following systemic NMDA receptor blockade. This, in turn, suggests that NMDA receptor activation may be an important component of the neural circuitry involved in satiation. PMID- 9855489 TI - Visual alliesthesia--cloth color preference in the evening under the influence of different light intensities during the daytime. AB - The primary purpose of this study was to determine the effect of bright and dim light exposure during the daytime on cloth color preference in the evening. Ten healthy female volunteers were exposed to bright light of 5000 lx ("Bright") or dim light of 200 lx ("Dim") from 0730 +/- 1 h to 1800 hours, 200 lx from 1800 hours to 2330 +/- 1 h, and complete darkness during the sleep period (2330 hours +/- 1 h to 0730 +/- 1 h). The subjects were instructed to select the most preferable single one out of 41 cloth colors (24 x 52 cm, 100% cotton), every 5 min from 2100 hours to 2200 hours. Most subjects preferred a warmer cloth color in the Dim condition. The rectal temperature showed clear circadian rhythm both under Bright and Dim conditions but it was significantly lower during sleep in the Bright condition. On the contrary, leg skin temperature was significantly higher during the evening and sleep in the Bright. It is concluded that warmer cloth color is preferred in the evening after Dim condition at an ambient temperature, being based on the higher set point of core temperature after Dim. PMID- 9855490 TI - Restricted lesions to ventral prefrontal subareas block reversal learning but not visual discrimination learning in rats. AB - Previous studies have shown that extensive damage to the medial prefrontal cortex (mPFC) of rats causes reversal learning deficits. The mPFC of rats, however, consists of several subareas that are different from each other in both cytoarchitecture and neural connectivity, suggesting a functional dissociation among the mPFC subareas. In the present study, selective lesions of the mPFC of rats were made with a specially designed microknife whose intracranial placement could be controlled stereotaxically. Restricted lesions were made to each of the 3 parts of the mPFC: the anterior cingulate area (AC) (including the medial precentral area, PrCm), the prelimbic area (PL), and the infralimbic area (IL). One week after surgery, rats were trained in an aversively motivated visual discrimination task in a novel rotating T-maze. After reaching the acquisition criterion, rats were trained in a reversal task in the same maze. No difference was found in acquisition between control and mPFC lesioned rats. However, lesions of either the PL or the IL produced a marked deficit in the reversal task. This behavioral deficit was not found in rats with lesions of the AC. The results indicate that the mPFC of rats is not essential for discrimination learning, but that each of the 2 ventral subareas of the mPFC, PL, and IL, plays a critical role in reversal learning. PMID- 9855491 TI - Purkinje cell responses to pontine stimulation CS during rabbit eyeblink conditioning. AB - Previous studies have shown that stimuli typically used as CSs in eyeblink conditioning converge with US information in the cerebellum. Extracellular recordings of Purkinje cells have shown learning-related as well as stimulus related discharge patterns. Stimulation of a portion of the auditory CS pathway, the pontine nucleus, also serves as a highly effective CS. Using a short-latency pontine stimulation CS and air puff US, single Purkinje cell responses were recorded and compared to those elicited with an auditory stimulus in previous work. Purkinje cell recordings in trained and untrained rabbits revealed patterns of responses very similar to those seen in rabbits trained to a tone CS or those given unpaired-tone/air-puff training. Similarities included the proportion of stimulus-related and behavior-related cell responses. However, fewer inhibitory responses were seen than in earlier studies and these differences are considered in light of the differences between an extracellular stimulation CS and a peripherally administered auditory CS. PMID- 9855492 TI - Sex differences in spontaneous locomotor activity and rotational behavior in meadow voles. AB - Sex differences in rotational behavior have been most clearly established in laboratory rats with females exhibiting a turning bias. Here, using an automated open-field apparatus, locomotor activity and spontaneous rotational behavior were examined in diurnally crepuscularly active reproductive male and female meadow voles (Microtus pennsylvanicus). Meadow voles, being induced ovulators, permitted analysis of females in constant behavioral estrous. Males displayed significantly greater levels of activity and also significantly greater levels of clockwise but not counterclockwise rotational behavior relative to the females. Rotational behavior was less strongly related to activity levels in female as compared to male voles. In addition, females displayed an overall turning bias. These results contrast with findings from laboratory rats in which females are reported to display greater levels of both locomotor activity and rotational behavior. They are, however, consistent with the rotational bias evident in female rats. The present findings confirm the presence of sex differences in rotational behavior and indicate that factors other than activity levels are involved in the generation and/or expression of these sex differences. Sex differences in anxiety and routine-like behavior (i.e., asymmetry in movement) are discussed as possible factors contributing to these male-female differences in rotational behavior. PMID- 9855493 TI - Aspartame consumption in rats selectively bred for high versus low saccharin intake. AB - Whereas humans use aspartame as a sugar substitute, evidence to date from rats suggests that aspartame does not taste sweet or, more generally, hedonically positive to them. The present study provided a strong test of the appetitive properties of aspartame in rats by examining consumption of aspartame and, for comparison, several sugars by two lines of rats selectively bred for high (HiS) versus low (LoS) saccharin consumption. The HiS and LoS lines differed in consumption of fructose, glucose, sucrose, maltose, and saccharin solutions. Overall, the rats showed a weak but significant preference for aspartame. However, no line differences in aspartame consumption were observed. Thus, even among rats specifically bred on the basis of their responsiveness to sweet tastes, aspartame tastes minimally sweet or good. PMID- 9855494 TI - Deletions of mitochondrial DNA in Kearns-Sayre syndrome. 1988. PMID- 9855495 TI - Avoiding conflicts of interest: responsibilities of authors, reviewers, and editors. PMID- 9855496 TI - Serotonin neurotoxicity: implications for cognitive neuroscience and neurology. PMID- 9855497 TI - Even the dead are not terminally ill anymore. PMID- 9855498 TI - Memory impairment in abstinent MDMA ("Ecstasy") users. AB - BACKGROUND: Methylenedioxymethamphetamine (MDMA, or "Ecstasy") is a popular recreational drug of abuse that is known to damage brain serotonergic neurons in animals and possibly humans. Few functional consequences of MDMA-induced serotonin (5-HT) neurotoxicity have been identified, either in animals or humans. This study sought to determine whether individuals with a history of extensive MDMA use showed evidence of memory impairment, because brain serotonin has been implicated in mnemonic function. METHOD: The authors compared 24 abstinent MDMA users and 24 control subjects on several standardized tests of memory, after matching subjects for age, gender, educational level, and vocabulary score (a surrogate of verbal intelligence). The authors also explored correlations between changes in memory function and decrements in CSF 5-hydroxyindoleacetic acid (5 HIAA), which serves as a marker of central 5-HT neural function. RESULTS: Greater use of MDMA (total milligrams per month) was associated with greater impairment in immediate verbal memory (p < 0.02) and delayed visual memory (p < 0.06). Furthermore, lower vocabulary scores were associated with stronger dose-related effects, with men having greater dose-related deficits than women. Lastly, lower concentrations of CSF 5-HIAA were associated with poorer memory performance. CONCLUSION: Abstinent MDMA users have impairment in verbal and visual memory. The extent of memory impairment correlates with the degree of MDMA exposure and the reduction in brain 5-HT, as indexed by CSF 5-HIAA. PMID- 9855499 TI - Chronic "brain death": meta-analysis and conceptual consequences. AB - OBJECTIVE: One rationale for equating "brain death" (BD) with death is that it reduces the body to a mere collection of organs, as evidenced by purported imminence of asystole despite maximal therapy. To test this hypothesis, cases of prolonged survival were collected and examined for factors influencing survival capacity. METHODS: Formal diagnosis of BD with survival of 1 week or longer. More than 12,200 sources yielded approximately 175 cases meeting selection criteria; 56 had sufficient information for meta-analysis. Diagnosis was judged reliable if standard criteria were described or physicians made formal declarations. Data were analyzed by means of Kaplan-Meier curves, with treatment withdrawals as "censored" data, compared by log-rank test. RESULTS: Survival probability over time decreased exponentially in two phases, with initial half-life of 2 to 3 months, followed at 1 year by slow decline to more than 14 years. Survival capacity correlated inversely with age. Independently, primary brain pathology was associated with longer survival than were multisystem etiologies. Initial hemodynamic instability tended to resolve gradually; some patients were successfully discharged on ventilators to nursing facilities or even to their homes. CONCLUSIONS: The tendency to asystole in BD can be transient and is attributable more to systemic factors than to absence of brain function per se. If BD is to be equated with death, it must be on some basis more plausible than loss of somatic integrative unity. PMID- 9855500 TI - Frontotemporal lobar degeneration: a consensus on clinical diagnostic criteria. AB - OBJECTIVE: To improve clinical recognition and provide research diagnostic criteria for three clinical syndromes associated with frontotemporal lobar degeneration. METHODS: Consensus criteria for the three prototypic syndromes frontotemporal dementia, progressive nonfluent aphasia, and semantic dementia were developed by members of an international workshop on frontotemporal lobar degeneration. These criteria build on earlier published clinical diagnostic guidelines for frontotemporal dementia produced by some of the workshop members. RESULTS: The consensus criteria specify core and supportive features for each of the three prototypic clinical syndromes and provide broad inclusion and exclusion criteria for the generic entity of frontotemporal lobar degeneration. The criteria are presented in lists, and operational definitions for features are provided in the text. CONCLUSIONS: The criteria ought to provide the foundation for research work into the neuropsychology, neuropathology, genetics, molecular biology, and epidemiology of these important clinical disorders that account for a substantial proportion of cases of primary degenerative dementia occurring before the age of 65 years. PMID- 9855501 TI - Brain volume preserved in healthy elderly through the eleventh decade. AB - OBJECTIVE: To determine which brain regions lose volume with aging over time in healthy, nondemented elderly. BACKGROUND: Cross-sectional studies suggest widespread loss of brain volume with aging. These studies may be biased by significant numbers of preclinically demented elderly in the oldest comparison groups. Longitudinal studies may allow closer determination of the effect of aging unaffected by dementia. METHODS: Quantitative volumetric MRI was performed annually on 46 healthy subjects older than age 65 who had maintained cognitive health a mean of 5 years. Comparisons (analysis of variance) were made of rates of volume loss (slopes) divided into 11 young-old (mean age, 70 years), 15 middle old (mean age, 81 years), and 20 oldest-old (mean age, 87 years) subjects. Regions of interest included CSF spaces, lobar regions, and limbic-subcortical regions. RESULTS: There were significant differences between groups in intracranial, total brain, left hemisphere, right hemisphere, temporal lobe, basilar-subcortical region, and hippocampus volumes, with oldest-old subjects showing the smallest volumes, followed by middle-old and young-old subjects. Oldest-old subjects had significantly greater subarachnoid volumes than the younger groups. There were no significant differences in rates of change of regions of interest across age groups. CONCLUSIONS: After age 65 there is minimal brain volume loss observed over time in healthy elderly. Brain volume differences seen cross-sectionally, at any age, likely reflect small, constant rates of volume loss with healthy aging. Healthy oldest-old subjects do not show greater rates of brain loss compared with younger elderly, suggesting that large changes seen in cross-sectional studies reflect the presence of preclinical dementia in older groups. PMID- 9855502 TI - Decreased glutathione transferase activity in brain and ventricular fluid in Alzheimer's disease. AB - OBJECTIVE: To investigate the levels of glutathione transferase (GST), a protective enzyme against aldehydes, and especially 4-hydroxynonenal (HNE) in the brain and ventricular CSF of autopsied AD and normal control subjects. BACKGROUND: Studies have implicated increased levels of oxidative stress in the brain in the pathogenesis of AD. Decreased levels of polyunsaturated fatty acids and increased levels of markers of lipid peroxidation have been reported in the brain in AD, particularly in areas severely affected in the disease. HNE, one marker of lipid peroxidation, is neurotoxic in neuronal culture and in vivo and is elevated in AD brain and CSF. METHODS: We measured levels of GST activity and protein in multiple brain regions and ventricular CSF in short-postmortem interval AD patients and age-matched prospectively evaluated control subjects. RESULTS: A decrease in GST activity in all brain areas was observed in AD compared with controls with significant decreases in the amygdala, hippocampus and parahippocampal gyrus, inferior parietal lobule, and nucleus basalis of Meynert. Levels of GST protein also were depleted in most brain regions in AD. A significant decrease in GST activity and protein levels was also found in ventricular CSF in AD. CONCLUSION: Reduced levels of GST, a protective mechanism against HNE, may have a role in the pathogenesis of neuron degeneration in AD. PMID- 9855503 TI - Exploring the temporal nature of hemodynamic responses of cortical motor areas using functional MRI. AB - OBJECTIVE: To use functional MRI (fMRI) to study grouped patterns of cerebral activation and the course of hemodynamic responses during performance of two activation tasks (paradigms) using a hand-held joystick to perform movements in a repetitively fixed direction and movements in freely selected random directions. BACKGROUND: Evidence from lesion, electrophysiologic, and functional imaging studies implicates prefrontal and mesial frontal cortex in motor preparation and primary motor cortex in motor execution. fMRI can be used to study cerebral activation and has practical advantages over other methods of functional neuroimaging. METHODS: We acquired 100 multislice T2*-weighted data sets from five healthy volunteers during performance of each paradigm using conventional fMRI. For each paradigm, rest and movement epochs were alternated every 30 seconds. After coregistration and spatial normalization, we combined the data for group studies. We used statistical parametric mapping to compare the early (first 15 seconds) components of the movement epochs with rest as well as the late (last 15 seconds) components of the movement epochs with rest. RESULTS: During the early phase of both paradigms, significant activation was present in rostral and caudal mesial premotor cortex. Right prefrontal cortex was significantly activated during the early component of freely selected joystick movements. Activation of rostral supplementary motor area was maintained during the late component of freely selected movements but decreased during repetitively fixed movements. In contrast, significant activation in contralateral sensorimotor cortex was maintained during both early and late components of both paradigms. CONCLUSIONS: fMRI can detect cortical activation. The temporal resolution of fMRI also allows adaptation of blood oxygenation level-dependent (BOLD) contrast signal to be detected in association cortex. However, the level of BOLD contrast signal in primary motor cortex remained significantly elevated throughout task performance. PMID- 9855504 TI - Cerebral blood flow velocity decreases during cognitive stimulation in Huntington's disease. AB - OBJECTIVE: To examine whether activation of cerebral blood flow velocity during cognitive stimulation, as measured by transcranial Doppler ultrasonography (TCD), is altered in patients with Huntington's Disease (HD). BACKGROUND: Previous research suggests that resting cerebral blood flow in symptomatic and asymptomatic HD patients is reduced from expected premorbid levels. The effects of cognitive activation on this relative hypoperfusion in HD has not been studied extensively. METHODS: We measured TCD flow velocity during rest and cognitive testing with the Porteus Maze Test and the Trails Test in 12 normal control subjects and 10 gene-positive HD patients. Percent change (i.e., flow during testing/resting) of flow velocity in the anterior and middle cerebral arteries were compared between groups. Correlations among percent flow velocity change, a disability rating scale, and cognitive test scores were calculated. RESULTS: In control subjects, anterior cerebral artery flow velocity and middle cerebral artery velocity increased during cognitive testing (p=0.001). HD patients showed a smaller blood flow velocity increase in the anterior cerebral arteries during the Porteus Maze Test (p < 0.001) and the Trails Test, Part B (p < 0.001). In the left anterior cerebral artery, flow velocity fell an average of 4.2% below resting baseline levels during the Porteus Maze Test, and 1.2% below resting levels during the Trails Test. The magnitude of the cerebral blood flow velocity changes in the anterior cerebral artery correlated with a number of cognitive test scores and with a rating scale of functional disability. In addition, logistic regression was able to discriminate the HD patients from the control group based on blood flow velocity changes (p=0.0025). When HD patients were divided into more (i.e., HD with chorea; n=4) and less impaired (i.e., without chorea; n=6) groups, both showed significant decreases in left anterior cerebral artery flow velocity during visual spatial executive cognition testing compared with control subjects. CONCLUSIONS: These results suggest that activation of visual spatial executive functions cause decreased flow velocity in the anterior cerebral artery, but not the middle cerebral artery, in HD patients. These changes are related to test performance and functional capabilities. PMID- 9855505 TI - Enhanced fatigue during motor performance in patients with Parkinson's disease. AB - OBJECTIVE: To measure fatigue quantitatively during continuous motor performance in patients with PD. BACKGROUND: Enhanced fatigue on performance of motor tasks is a very frequent and disabling complaint of PD patients, and is poorly characterized and understood. Recent evidence suggests a role for mitochondrial dysfunction in the pathogenesis of PD. Reduced exercise capacity is one of the hallmarks of systemic mitochondrial impairment. METHODS: The authors used an automated system to measure muscle fatigue during a continuous (30-second), maximal, isometric forearm flexion in 17 PD patients and 10 age-matched control subjects. A fatigue index (FI) was then calculated. Peak force (PF) was measured as an internal standard of the examination. Measurements were performed before and 2 hours after an oral dose of levodopa/carbidopa (125 mg/12.5 mg). RESULTS: In PD patients there was a 50% increase in FI. The increased FI was often asymmetric and more pronounced on the side more affected by the disease. FI was significantly responsive to, and improved after, an oral dose of levodopa. The rate of improvement in FI induced by levodopa correlated with disease severity, as measured by the Unified Parkinson's Disease Rating Scale. No significant alterations in PF were observed. CONCLUSIONS: Enhanced muscle fatigue should be recognized as an integral part of the spectrum of motor impairment of PD. However, our data argue for its association with a central dopamine deficiency rather than with a muscle mitochondrial abnormality. PMID- 9855506 TI - Slow allotypic variants of the NAT2 gene and susceptibility to early-onset Parkinson's disease. AB - OBJECTIVE: To determine the frequency and the linkage distribution of seven mutations at the polymorphic gene coding for the arylamine N-acetyl transferase (NAT2; EC 2.3.1.5) in 121 unrelated patients with sporadic PD and in 121 unrelated healthy volunteers. METHODS: The study was performed with mutation specific PCR using genomic DNA obtained from blood of the probands. RESULTS: Comparison of the NAT2 genotypes of the overall PD patients and control subjects did not indicate statistically significant differences. However, patients with early-onset PD (onset before the age of 50 years, n=37) showed a higher frequency of slow-acetylation genotypes (78.4% patients) compared with both healthy control subjects (55.4%) and with late-onset (onset after 51 years of age, n=84) PD patients (54.8%). Such a difference was statistically significant (p < 0.015) and was the result of a homogeneous increase in the frequency of slow-acetylation alleles. All subgroups analyzed in the study were in Hardy-Weinberg equilibrium for mutations at the NAT2 gene. CONCLUSIONS: Slow-acetylation-mutated alleles may be considered low-penetrance genes in early-onset PD pathogenesis, with a relative risk ratio for individuals with slow-acetylation genotype of 2.92 (95% CI, 1.26 to 6.78). This study provides evidence for the interaction of genetic and environmental factors in the etiology of sporadic PD. PMID- 9855507 TI - Thyroarytenoid muscle activity associated with hypophonia in Parkinson disease and aging. AB - OBJECTIVE: To compare electromyographic (EMG) amplitudes of the thyroarytenoid (TA) muscle in young and older individuals and individuals with idiopathic PD (IPD) under conditions of known vocal loudness (sound pressure level). BACKGROUND: Voice disorders frequently accompany aging and IPD, but it is unclear how laryngeal muscle physiology is affected by these processes and how changes in laryngeal muscle activity result in characteristic changes of the voice. METHODS: Absolute and relative (to maximum) EMG amplitudes of the TA muscle were compared during speech and nonspeech tasks. Corresponding sound pressure level (SPL) measures were obtained for the speech tasks. RESULTS: Absolute TA amplitudes were consistently the highest in the young individuals, lowest in the individuals with IPD, and intermediate in the older individuals. Relative TA amplitudes were generally the highest for the young individuals, lowest for the older individuals, and intermediate for the individuals with IPD. SPL findings showed the older individuals and individuals with IPD produced most of the speech tasks with comparable SPLs, and these levels were consistently lower than those of the young individuals. CONCLUSIONS: Reduced levels of TA muscle activity may contribute to the characteristic hypophonic voice disorders that frequently accompany IPD and aging. PMID- 9855508 TI - Randomized, double-blind, placebo-controlled trial of pergolide in restless legs syndrome. AB - OBJECTIVE: To assess the efficacy of pergolide in the treatment of restless legs syndrome (RLS). METHODS: A randomized, double-blind, parallel-treatment evaluation of pergolide versus placebo in patients with RLS was performed. Sixteen patients seen in the Sleep Disorders Center who met the clinical criteria for RLS were enrolled in the study. Treatment was either placebo (n=8) or pergolide (n=8), which was on a self-adjusted schedule that ranged from 0.05 to 0.65 mg/day. Outcome variables included the patient's rating of symptom improvement, duration of RLS symptoms throughout the day, and objective sleep assessments from an all-night polysomnogram (percent sleep efficiency and periodic leg movements). Baseline was assessed for 2 days before starting treatment. Dose adjustments were made over the first 14 days and were then held constant for 5 days, with repeat assessment for treatment effects on the last 2 days of treatment (days 17 and 18). RESULTS: As a group, pergolide recipients showed significant improvement clinically and statistically in all outcome measures when compared with baseline or with placebo treatment. On the other hand, placebo treatment led to no significant improvements in any outcome measures. CONCLUSIONS: Pergolide treatment significantly improved clinical symptoms and sleep efficiency, and reduced periodic limb movements of sleep in patients with RLS. PMID- 9855509 TI - Sleep disturbance in spinocerebellar ataxias: is the SCA3 mutation a cause of restless legs syndrome? AB - OBJECTIVE: To identify and to characterize sleep disturbances in patients with hereditary ataxias. BACKGROUND: We observed restless legs syndrome (RLS) and impaired sleep as a frequent yet unrecognized symptom in spinocerebellar ataxia type 3 (SCA3). METHODS: A total of 89 patients with genetically defined subtypes of autosomal dominant cerebellar ataxias were investigated for sleep history and neurologic findings according to a standardized protocol. Nerve conduction studies were performed. Sleep was studied by overnight polysomnography in seven patients. RESULTS: RLS was present in 45% of SCA3 patients but is rare in other types of autosomal dominant cerebellar ataxias. RLS was a frequent but not the only cause of sleep impairment in SCA3. Impaired sleep in SCA3 is associated with older age, long-standing disease, and brainstem involvement. RLS tended to be more frequent in patients with clinical signs of polyneuropathy, but RLS was not restricted to patients with peripheral neuropathy. RLS was not observed in healthy members of SCA3 families. CONCLUSIONS: RLS is a frequent and treatable cause of disabling sleep disturbance in SCA3. This study provides evidence for the expanded CAG repeat in the SCA3 gene as a molecular factor causing RLS. PMID- 9855510 TI - Centrotemporal spikes in families with rolandic epilepsy: linkage to chromosome 15q14. AB - OBJECTIVE: To localize a gene predisposing to benign epilepsy of childhood with centrotemporal spikes (BECTS). BACKGROUND: BECTS, or rolandic epilepsy, is the most prevalent idiopathic epilepsy syndrome in childhood. Functional relevant defects in the alpha 4 subunit of the neuronal nicotinic acetylcholine receptor (AChR) have been demonstrated in autosomal dominant nocturnal frontal lobe epilepsy, which, like BECTS, is an idiopathic partial epilepsy. METHODS: A DNA linkage study was conducted screening all chromosomal regions known to harbor neuronal nicotinic AChR subunit genes. Twenty-two nuclear families with BECTS were analyzed. RESULTS: In an "affected-only" study, best p values and lod scores were reached between D15S165 and D15S1010 on chromosome 15q14. In multipoint nonparametric linkage analysis a nominal p value of 0.000494 was calculated by GENEHUNTER. Best parametric results were obtained under an autosomal recessive model with heterogeneity (multipoint lod score 3.56 with 70% of families linked to the locus). These markers are localized in direct vicinity to the alpha 7 subunit gene of the AChR. CONCLUSIONS: We found evidence for linkage of BECTS to a region on chromosome 15q14. Either the alpha 7 AChR subunit gene or a closely linked gene are implicated in pedigrees with BECTS. The disorder is genetically heterogeneous. Surprisingly, the same chromosomal area has been reported to be linked to the phenotype in families with an auditory neurophysiologic deficit as well as in families with juvenile myoclonic epilepsy, another idiopathic but generalized epilepsy syndrome. PMID- 9855511 TI - Wallerian degeneration of the pyramidal tract does not affect stroke rehabilitation outcome. AB - OBJECTIVE: To test whether Wallerian degeneration (WD) of the pyramidal tract as signaled by MRI affects rehabilitation outcome in patients with subcortical infarction (internal capsule or corona radiata). BACKGROUND: Recent radiologic evidence suggests that WD occurs no earlier than 3 months after a subcortical infarction. METHODS: A total of 77 consecutive patients with pure motor hemiparesis due to an initial subcortical infarction were assessed on admission and discharge with the Functional Independence Measure (FIM) for disability and Stroke Impairment Assessment Set (SIAS, full = 25) for impairment. WD was defined by a high-intensity area detected along the pyramidal tract below the level of lesion on T2-weighted MR image (WD+). RESULTS: Age, sex, side of stroke, Mini Mental State Examination score, and volume of lesion were comparable for each group. Length of stay (LOS) was significantly longer (p < 0.05) in WD+ (130 days) than in WD- (105 days). There was no difference in the change of FIM (WD+, 99 to 111; WD-, 95 to 107) or SIAS measures (WD+, 12 to 16; WD-, 13 to 16) made on admission and discharge, nor was there any effect of the timing of the rehabilitation experience (< or = 90 days or >90 days after stroke). CONCLUSIONS: After stroke, apparent WD of the pyramidal tract may slow functional recovery but does not limit final rehabilitation outcome of pure motor hemiparesis. Study of the mechanisms of compensation for this delayed pyramidal tract degeneration will enhance the scientific basis for rehabilitation. PMID- 9855512 TI - Identifying hypoxic tissue after acute ischemic stroke using PET and 18F fluoromisonidazole. AB - OBJECTIVE: To show that PET with 18F-fluoromisonidazole (18F-FMISO) can detect peri-infarct hypoxic tissue in patients after ischemic stroke. BACKGROUND: PET with (15)O-labeled oxygen and water is the only established method for identifying the ischemic penumbra in humans. We used PET with 18F-FMISO in patients after ischemic stroke to identify hypoxic but viable peri-infarct tissue likely to represent the ischemic penumbra, and to determine how long hypoxic tissues persist after stroke. METHODS: Patients with acute hemispheric ischemic stroke were studied using PET with 18F-FMISO either within 48 hours or 6 to 11 days after stroke onset. The final infarct was defined by CT performed 6 to 11 days after stroke. Tracer uptake was assessed objectively by calculating the mean activity in the contralateral (normal) hemisphere, then identifying pixels with activity greater than 3 SDs above the mean in both hemispheres. Positive studies were those with high-activity pixels ipsilateral to the infarct. RESULTS: Fifteen patients were studied; 13 within 48 hours of stroke, 8 at 6 to 11 days, and 6 during both time periods. Hypoxic tissue was detected in 9 of the 13 patients studied within 48 hours of stroke, generally distributed in the peripheries of the infarct and adjacent peri-infarct tissues. None of the 8 patients studied 6 to 11 days after stroke exhibited increased 18F-FMISO activity. All 6 patients studied both early and late exhibited areas of increased activity during the early but not the late study. CONCLUSIONS: PET with 18F-FMISO can detect peri infarct hypoxic tissue after acute ischemic stroke. The distribution of hypoxic tissue suggests that it may represent the ischemic penumbra. Hypoxic tissues do not persist to the subacute phase of stroke (6 to 11 days). PMID- 9855513 TI - Stroke and intracranial venous thrombosis during pregnancy and puerperium. AB - OBJECTIVES: To determine nationally representative estimates of the incidence of stroke and intracranial venous thrombosis during pregnancy and the puerperium, and to identify potential risk factors for these conditions. METHODS: National Hospital Discharge Survey data were analyzed for the period 1979 to 1991. Nationally representative estimates of risk were calculated by age, race, presence of pregnancy-related hypertension, census region, hospital ownership, and number of hospital beds. Multivariate models were developed using logistic regression. RESULTS: There were an estimated 8,918 cases of stroke and 5,723 cases of intracranial venous thrombosis during pregnancy and the puerperium in the United States among 50,264,631 deliveries, giving risks of 17.7 cases of stroke and 11.4 cases of intracranial venous thrombosis per 100,000 deliveries. In the multivariate models, stroke was associated strongly with pregnancy-related hypertension, larger hospital size, and proprietary hospital ownership, and inversely associated with living in the South. Intracranial venous thrombosis was associated with maternal age. CONCLUSIONS: Stroke and intracranial venous thrombosis are relatively common complications of pregnancy and the puerperium. Collectively, rates for these conditions are about 50% greater for the entire period of pregnancy and the puerperium than for the immediate peripartum period. PMID- 9855514 TI - Efficacy and safety of oral sildenafil (Viagra) in men with erectile dysfunction caused by spinal cord injury. AB - OBJECTIVE: To evaluate the efficacy and safety of 50-mg doses of sildenafil during a 28-day period in patients with erectile dysfunction caused by spinal cord injury (cord level range, T6 through L5). BACKGROUND: Sildenafil is an orally active, potent, and selective inhibitor of phosphodiesterase type 5, an important regulator of cyclic guanosine monophosphate in the human corpus cavernosum. METHODS: To be included in this double-blind, placebo-controlled study, all patients had to be able to achieve at least a partial reflexogenic erectile response to penile vibratory stimulation. The study utilized a single triangular sequential trial design. A total of 27 patients were randomized to receive 50 mg of sildenafil or placebo, taken orally as required (not more than once daily) approximately 1 hour before sexual activity. RESULTS: After 28 days of treatment, nine of 12 patients (75%) on sildenafil and one of 14 patients (7%) on placebo reported that treatment had improved their erections (p=0.0043). Furthermore, eight of 12 patients (67%) on sildenafil and two of 13 patients (15%) on placebo indicated that they wished to continue treatment (p=0.018). A significant improvement in satisfaction with their sex life was reported by patients taking sildenafil (p=0.012). No patients discontinued treatment due to adverse events. CONCLUSION: Oral sildenafil, taken as required (not more than once daily), significantly improves the quality of erections and satisfaction with sex life in men with erectile dysfunction caused by a spinal cord injury between T6 and L5. PMID- 9855516 TI - Recovery of nerve conduction following microvascular decompression for trigeminal neuralgia. AB - OBJECTIVE: To assess the function of trigeminal nerve before and after microvascular decompression for trigeminal neuralgia. BACKGROUND: To date there is no direct evidence that microvascular decompression of the trigeminal root restores normal conduction in the nerve. METHODS: The authors examined 10 patients with trigeminal neuralgia in whom preoperative MRI and MR angiography demonstrated neurovascular contact. During microvascular decompression, the trigeminal nerve was monitored by recording early scalp trigeminal evoked potentials immediately before, during, and after decompression. Direct recordings from the root entry zone were also performed. RESULTS: In all patients preoperative scalp evoked potentials showed impaired conduction of the trigeminal root. Microvascular decompression was associated with immediate recovery of conduction in seven patients, demonstrated by both scalp evoked potentials and direct root recordings. All 10 patients were pain free postoperatively. CONCLUSIONS: Improvement in trigeminal neuralgia following microvascular decompression is often associated with normalization of neurophysiologic data, suggesting recovery of nerve function. Rapid electrophysiologic recovery and pain relief following microvascular decompression argue that neither phenomenon is linked to remyelination. It is possible that the trigeminal evoked potentials might predict an effective microvascular decompression. PMID- 9855515 TI - Calciphylaxis mimicking dermatomyositis: ischemic myopathy complicating renal failure. AB - BACKGROUND: Among the complications of chronic renal failure is a syndrome of medial calcification of small- to medium-sized arteries associated with ischemic necrosis of the skin and other organ systems, leading to gangrene and a poor prognosis. The syndrome has been reviewed in the renal, dermatologic, and surgical literature under the term calciphylaxis, which describes a postulated pathogenetic mechanism whereby sensitization to an endogenous or exogenous substance (such as parathyroid hormone) predisposes to calcium deposition after exposure to a challenging agent. Myopathy has rarely been reported as the presenting feature, and the syndrome has not been discussed in the neurologic literature. METHODS: We report two patients with renal failure and systemic calciphylaxis who presented to our hospital with myopathic complaints and signs suggesting dermatomyositis. We also discuss possible disease mechanisms and treatment. CONCLUSIONS: Because early treatment (including aggressively lowering the calcium and phosphate levels and parathyroidectomy) may improve the outcome, early recognition of the syndrome of calciphylaxis is essential. PMID- 9855517 TI - The wide spectrum of myofibrillar myopathy suggests a multifactorial etiology and pathogenesis. AB - BACKGROUND: Myofibrillar myopathy (MFM) is characterized by nonhyaline lesions (foci of myofibrillar destruction) and hyaline lesions (cytoplasmic inclusions composed of compacted myofibrillar residues) on light and electron microscopy. Immunocytochemistry demonstrates the abnormal expression of desmin and numerous other proteins. The clinical, laboratory, and histologic features of MFM are heterogeneous, making a diagnosis difficult. RESULTS: We diagnosed eight patients with MFM over the preceding 3 years. MFM was inherited in an autosomal dominant pattern in one patient, developed sporadically in five patients, and was induced by an experimental chemotherapy, Elinafide (Knoll, Parsippany, NJ), in two patients. Age at onset ranged from 14 to 64 years. The pattern of weakness was variable but involved proximal and distal muscles. Five patients had evidence of a cardiomyopathy. Electromyography demonstrated muscle membrane instability and small, polyphasic motor unit potentials. Serum creatine kinase levels were normal to moderately elevated (<10x normal). Light and electron microscopy demonstrated the characteristic pattern of nonhyaline and hyaline lesions and the associated abnormalities on immunocytochemistry. CONCLUSIONS: Patients demonstrate a wide spectrum of clinical, laboratory, and histologic abnormalities. Chemotherapy induced MFM has abnormalities on immunocytochemistry similar to the those of hereditary and sporadic cases. The pathogenesis of MFM is likely heterogeneous. However, MFM is distinctive in that it can preferentially affect distal muscles and has a frequent association with cardiomyopathy. The cardiomyopathy may be amenable to treatment with pacemaker insertion or cardiac transplantation. PMID- 9855518 TI - Two patterns of clinical recovery in Guillain-Barre syndrome with IgG anti-GM1 antibody. AB - OBJECTIVE: To investigate the prognostic value of anti-GM1 antibody. BACKGROUND: Whether anti-GM1 antibody is a marker of poor prognosis due to axonal degeneration in Guillain-Barre syndrome (GBS) is a matter of controversy. METHODS: The clinical recovery of 41 consecutive GBS patients was analyzed. RESULTS: The Hughes functional grading scores were similar at the peak, and 1, 3, and 6 months after onset for the groups of patients with (n=19) and without (n=22) immunoglobulin (Ig) G anti-GM1 antibodies. However, the anti-GM1-positive group included significantly higher proportions of patients with poor recovery (inability to walk independently at 6 months, 5 of 19 versus 0 of 22; p=0.01) and those with a markedly rapid recovery (improvement by two or more Hughes grades within a month, 9 of 19 versus 4 of 22; p=0.05). The positivity of IgG anti-GM1 antibody correlated well with the electrodiagnosis of the acute motor axonal neuropathy pattern but was not always associated with poor prognosis. Anti-GM1 positive patients showed two different patterns of clinical recovery-their conditions improved slower or faster than those of the anti-GM1-negative patients, most of whom had acute inflammatory demyelinating polyneuropathy. CONCLUSIONS: Anti-GM1 antibody is not always a marker of poor prognosis and, besides axonal degeneration, early reversible effects other than demyelination could be part of the pathophysiology of Guillain-Barre syndrome with IgG anti-GM1 antibody. PMID- 9855519 TI - Inflammatory cytokines inhibit upregulation of glycolipid expression by Schwann cells in vitro. AB - OBJECTIVE: To determine whether products of inflammatory cells can inhibit differentiation and synthesis of myelin glycolipids by Schwann cells. BACKGROUND: Infiltration of the peripheral nervous system by inflammatory cells is a feature of acquired demyelinating neuropathies. It is not clear what role these cells have in causing demyelination or inhibiting myelin synthesis. METHODS: Nonmyelinating rat Schwann cells were incubated with 1) different concentrations of activated supernatants (AS) from mitogen-activated inflammatory cells; 2) 8 bromo cyclic adenosine monophosphate (8Br cAMP), known to induce Schwann cell differentiation and synthesis of glycolipids; 3) 8Br cAMP and varying concentrations of AS; 4) 8Br cAMP and cytosine arabinoside (Ara C), which inhibits Schwann cell proliferation; 5) 8Br cAMP, AS, and Ara C; or 6) additional medium. RESULTS: AS inhibits the capacity of cAMP to induce Schwann cell expression of myelin-associated glycolipids. Inhibition of glycolipid expression was independent of the capacity of these AS to induce Schwann cell proliferation. CONCLUSIONS: These data suggest that inflammatory mediators are capable of inhibition of Schwann cell differentiation and synthesis of myelin. PMID- 9855520 TI - Incidence of dominant spinocerebellar and Friedreich triplet repeats among 361 ataxia families. AB - OBJECTIVE: To determine the incidence of spinocerebellar ataxia (SCA) types 1, 2, 3, 6, and 7 and Friedreich's ataxia (FA) among a large panel of ataxia families. BACKGROUND: The ataxias are a clinically and genetically heterogeneous group of neurodegenerative diseases that variably affect the cerebellum, brainstem, and spinocerebellar tracts. Trinucleotide repeat expansions have been shown to be the mutational mechanism for five dominantly inherited SCAs as well as FA. METHODS: We collected DNA samples and clinical data from patients representing 361 families with adult-onset ataxia of unknown etiology. Patients with a clinical diagnosis of FA were specifically excluded from our collection. RESULTS: Among the 178 dominant kindreds, we found SCA1 expansion at a frequency of 5.6%, SCA2 expansion at a frequency of 15.2%, SCA3 expansion at a frequency of 20.8%, SCA6 expansion at a frequency of 15.2%, and SCA7 expansion at a frequency of 4.5%. FA alleles were found in 11.4% of apparently recessive and 5.2% of apparently sporadic patients. Among these patients the repeat sizes for one or both FA alleles were relatively small, with sizes for the smaller allele ranging from 90 to 600 GAA repeats. The clinical presentation for these patients is atypical for FA, with one or more of the following characteristics: adult onset of disease, retained tendon reflexes, normal plantar response, and intact or partially intact sensory perceptions. CONCLUSIONS: Pathogenic trinucleotide repeat expansions were found among 61% of the dominant kindreds. Among patients with apparently recessive or negative family histories of ataxia, 6.8% and 4.4% tested positive for a CAG expansion at one of the dominant loci, and 11.4 and 5.2% of patients with apparently recessive or sporadic forms of ataxia had FA expansions. Because of the significant implications that a dominant versus recessive inheritance pattern has for future generations, it is important to screen patients who do not have a clearly dominant inheritance pattern for expansions at both the FA and the dominant ataxia loci. PMID- 9855521 TI - Olfactory dysfunction in Guamanian ALS, parkinsonism, and dementia. AB - OBJECTIVES: To assess whether olfactory deficits are present in the general Guamanian Chamorro population and to evaluate olfaction in each of the four neurodegenerative disease syndromes of Guam: ALS, pure parkinsonism, pure dementia, and the combined parkinsonism-dementia complex (PDC). BACKGROUND: Olfactory dysfunction was previously reported in patients with PDC of Guam. METHODS: We developed a culturally adjusted olfactory test battery, derived from the original University of Pennsylvania Smell Identification Test (UPSIT), and administered this to Chamorro residents with ALS (n=9), pure parkinsonism (n=9), pure dementia (n=11), PDC (n=31), and 53 neurologically normal Chamorro and 25 North American control subjects. RESULTS: Similar, marked olfactory dysfunction was found in all four syndromes of Guamanian neurodegenerative disease. This correlated poorly with measures of parkinsonism and cognition. In the neurologically normal Chamorro control group, six subjects (11%) had very low olfactory scores; these were less than the lowest North American score, raising a question of subclinical neurodegenerative disease. CONCLUSIONS: Marked olfactory deficits are common to all four Guamanian neurodegenerative syndromes, and suggest the possibility of similar central neuropathologic substrates. The deficit in the Guamanian ALS group contrasts with idiopathic ALS, in which olfactory function has been reported to be only slightly compromised. PMID- 9855522 TI - Screening for distal sensory peripheral neuropathy in HIV-infected persons in research and clinical settings. AB - OBJECTIVE: To determine the accuracy of a screening examination for distal sensory peripheral neuropathy (DSPN) performed by nonphysician clinicians and to explore the associations between DSPN and clinical features in HIV-infected persons. METHODS: A case-control study of a volunteer sample of 226 HIV-infected individuals was performed. An interview, focusing on risks and symptoms of DSPN, and a screening neurologic examination were performed. RESULTS: Compared with the neurologist's examinations, the clinicians' examination was sensitive (92 to 95%) but not as specific (71 to 84%) for the diagnosis of DSPN. After excluding 27 patients with confounders, 42 of 199 patients (21%) had DSPN. This was associated significantly with neurotoxic nucleoside antiretroviral use and with more advanced HIV disease. Of the 42 patients with DSPN, 30 (71%) had no neuropathy symptoms. CONCLUSIONS: A brief examination performed by trained nonphysician clinicians can be used to screen for DSPN in HIV-infected persons. Asymptomatic DSPN is common in these individuals. PMID- 9855524 TI - A serial study of new MS lesions and the white matter from which they arise. AB - OBJECTIVE: To compare MS normal-appearing white matter (NAWM) where new gadolinium-enhancing (Gd+) lesions do and do not arise. METHODS: A total of 22 relapsing-remitting MS patients and 11 healthy control subjects completed as many as 12 monthly brain MRI sessions. Quantitative measures of gadolinium enhancement (GDR), water proton density (PDN), water proton T2 relaxation time constants (T2), magnetization transfer ratio (MTR), and T1-weighted signal intensity (T1N) were followed serially in healthy control and MS NAWM. RESULTS: A total of 129 new Gd+ lesions were identified in 11 patients. PDN, T2, MTR, and T1N were diffusely abnormal in MS NAWM. NAWM regions in which new Gd+ lesions arose have increased GDR, PDN, and T2, and reduced MTR and T1N compared with contralateral homologous NAWM regions in which no new Gd+ lesions arose. Differences between these NAWM regions preceded lesion appearance for at least several months. After lesions became visible, GDR returned to baseline within 2 months, and PDN and MTR had larger residual abnormalities than T2 or T1N. CONCLUSIONS: Quantitative MRI measures are diffusely abnormal in MS NAWM. These measures are, on average, more abnormal in NAWM regions in which new Gd+ lesions arise. After the appearance of Gd+ lesions, measures of PDN and MTR may provide more appealing markers of relatively irreversible tissue damage than measures of T2 and T1N. PMID- 9855523 TI - A randomized trial of amitriptyline and mexiletine for painful neuropathy in HIV infection. AIDS Clinical Trial Group 242 Protocol Team. AB - BACKGROUND: Painful sensory neuropathy is a common complication of HIV infection. Based on prior uncontrolled observations, we hypothesized that amitriptyline or mexiletine would improve the pain symptoms. METHOD: A randomized, double-blind, 10-week trial of 145 patients assigned equally to amitriptyline, mexiletine, or matching placebo. The primary outcome measure was the change in pain intensity between baseline and the final visit. RESULTS: The improvement in amitriptyline group (0.31+/-0.31 units [mean+/-SD]) and mexiletine group (0.23+/-0.41) was not significantly different from placebo (0.20+/-0.30). Both interventions were generally well tolerated. CONCLUSIONS: Neither amitriptyline nor mexiletine provide significant pain relief in patients with HIV-associated painful sensory neuropathy. PMID- 9855525 TI - Neutralizing antibodies to interferon beta-1a and interferon beta-1b in MS patients are cross-reactive. AB - OBJECTIVE: To determine whether neutralizing antibodies (NABs) to interferon beta (IFNbeta)-1a (Avonex) and IFNbeta-1b (Betaseron) cross-react. BACKGROUND: A total of 38% of MS patients treated with IFNbeta-1b and 22% of those treated with IFNbeta-1a were reported to develop NABs, which could reduce the clinical efficacy of the drug. METHODS: Blood from 10 MS patients was collected before and at 3 and 6 months after initiating treatment with IFNbeta-1a. ELISA was performed to detect binding antibodies to IFNbeta-1a. Sera from patients who tested positive for binding antibodies to IFNbeta-1a were then screened for NABs to IFNbeta-1a in a biologic assay based on neutralization of antiviral activity. These serum samples were subsequently tested for cross-reactivity with IFNbeta-1b both in the ELISA and the biologic assay. In the second part of the study, sera from patients who participated in the phase III IFNbeta-1b trial at the University of Maryland were examined for cross-reactivity with IFNbeta-1a in the ELISA and the biologic assay. RESULTS: Of the 10 patients treated with IFNbeta 1a, three developed binding as well as NABs to IFNbeta-1a 6 months after treatment, and these antibodies cross-reacted with IFNbeta-1b both in the binding and the biologic assay. Similarly, sera from six patients with NABs to IFNbeta-1b showed cross-reactivity with IFNbeta-1a in the binding assay. Three of these six serum samples tested for neutralizing activity against IFNbeta-1a demonstrated the presence of NABs to IFNbeta-1a. CONCLUSIONS: NABs to IFNbeta-1a (Avonex) and IFNbeta-1b (Betaseron) cross-react, both in the binding and the biologic assays. This suggests that switching to alternate IFNbeta preparation in patients who develop NABs may not be clinically beneficial. Studies examining cross-reactivity between NABs to IFNbeta-1a and IFNbeta-1b in a large number of patients are indicated. PMID- 9855527 TI - Transient ischemic attacks caused by trumpet playing. AB - We report a young musician who experienced transient ischemic attacks during trumpet playing. Cardiovascular examination revealed a patent foramen ovale; Doppler ultrasound of the middle cerebral arteries detected microembolic signals with increased rates during trumpet playing and Valsalva maneuver. After operative occlusion of the foramen ovale, the symptoms disappeared and no microembolic signals could be detected. PMID- 9855528 TI - Lumbar and ventricular CSF protein, leukocytes, and lactate in suspected bacterial CNS infections. AB - Protein concentration, leukocyte density, and lactate concentration were studied in 41 pairs of ventricular and lumbar CSF drawn at an interval of less than 24 hours from patients with suspected bacterial CNS infections. The ventriculo lumbar ratios ranged from 0.003 to 10.2 (median=0.42) for protein and from 0.002 to 53.5 (median=0.17) for leukocytes. The uneven distribution of leukocytes and proteins in the CSF space may produce findings that fail to indicate bacterial CNS infections. Lactate was distributed more homogeneously in the CSF space than protein and leukocytes (ventriculo-lumbar ratio 0.52 to 1.66 [median=0.811). PMID- 9855526 TI - Methylprednisolone reduces adhesion molecules in blood and cerebrospinal fluid in patients with MS. AB - OBJECTIVE: To analyze the expression of adhesion molecules on mononuclear cells from blood and CSF of patients with exacerbations of MS before and after megadose IV methylprednisolone (MP). BACKGROUND: Adhesion molecules regulate transmigration of lymphocytes and monocytes/macrophages to the CNS and have an important role in the pathogenesis of MS. METHODS: The expression of very late activation antigen 4 (VLA-4) and vascular cell adhesion molecule 1, lymphocyte function-associated antigen 1 (LFA-1), and intercellular adhesion molecule 1 (ICAM-1) was analyzed immunocytologically on lymphocytes and monocytes from blood and CSF of 23 patients and 11 healthy control subjects. The results were correlated with the Expanded Disability Status Scale and in half of the patients with the number of T2-weighted MS plaques and brain atrophy analyzed by MRI. RESULTS: After treatment, the mean proportions of VLA-4, LFA-1, and ICAM-1 on blood lymphocytes (p < 0.0003, p < 0.00001, p < 0.01) and monocytes (p < 0.0001, p < 0.0002, p < 0.007) of 23 patients decreased. The expression of these adhesion proteins was also diminished on CSF leukocytes. However, even after treatment, the levels of VLA-4 and LFA-1 on lymphocytes from blood of MS patients remained higher than in the control subjects. The level of VLA-4 and LFA-1 on blood lymphocytes (r=0.67, p=0.023) and VLA-4 on monocytes (r=0.61, p=0.047) correlated with the number of T2-weighted lesions. CONCLUSIONS: Megadose MP may suppress brain inflammation by reducing the expression of adhesion molecules on mononuclear cells from blood and CSF of MS patients. The inhibition of cellular trafficking in MS by MP offers an important means of altering the autoimmune response in MS. PMID- 9855529 TI - Fatal familial insomnia: clinical and pathologic heterogeneity in genetic half brothers. AB - We describe clinical and pathologic features of a patient with fatal familial insomnia (FFI) whose prion (PrP) genotype is D178N coupled with methionine at codon 129 on his mutant allele and valine at codon 129 on his normal allele. A cousin (genetic half brother) with identical PrP genotypes exhibited strikingly different clinical and pathologic changes. Comparison of these cousins shows the phenotypic heterogeneity of FFI and suggests that the phenotypic expression of D178N is influenced by multiple factors. PMID- 9855530 TI - The natural history of asymptomatic meningiomas in Olmsted County, Minnesota. AB - To define the natural history of asymptomatic meningioma found incidentally on a neuroimaging study, we performed a retrospective analysis of 35 such patients. There were 32 women and three men, with a mean age of 67 years and a mean follow up of 74 months. Four tumors had progressed on subsequent imaging, and one patient developed symptoms related to the meningioma. Noncalcified tumors were more likely to progress than calcified tumors. PMID- 9855531 TI - Enhanced spasticity in primary progressive MS patients treated with interferon beta-1b. AB - Spasticity is a disabling symptom of MS that is enhanced during interferon beta lb (IFNbeta-1b) treatment. Nineteen patients with primary progressive MS were treated with IFNbeta-1b; an additional 19 patients did not receive this treatment. Thirteen of the 19 patients treated with IFNbeta-1b had increased spasticity requiring increased antispasticity drug administration. This observation suggests that further studies are needed before interferons can be so widely used in primary progressive MS patients. PMID- 9855532 TI - Seizure onset from periventricular nodular heterotopias: depth-electrode study. AB - The association between gray matter heterotopias and seizures is well established; whether seizures originate from these lesions is not known. We evaluated three patients with intractable complex partial seizures and periventricular nodular heterotopias (PNHs) with video-EEG monitoring with multiple depth electrodes, including placement in the PNH, to determine whether seizures originate from the PNH. In two of the three patients, all seizures arose from the PNH as low-voltage beta activity. In the third patient, 80% arose from the hippocampi and 20% from the heterotopia. PNHs may serve as an epileptogenic focus in patients with intractable epilepsy. PMID- 9855533 TI - Six-month evaluation of Carbatrol (extended-release carbamazepine) in complex partial seizures. AB - We evaluated the use of a new, controlled-release capsule form of carbamazepine, Carbatrol capsules, in an open-label, multicenter study of 124 patients with complex partial seizures. Ninety-one percent of the patients successfully completed the 6-month trial with good seizure control, with a significant improvement in quality of life. We conclude that switching patients with complex partial seizures from multiple daily-dose carbamazepine to twice-daily Carbatrol on a milligram-to-milligram basis is relatively safe. PMID- 9855534 TI - Evaluation of the alpha(2A)-adrenergic receptor gene in a heritable form of temporal lobe epilepsy. AB - An autosomal dominant form of human temporal lobe epilepsy (TLE) has been mapped to a region of chromosome 10q that contains the intronless alpha(2A)-adrenergic receptor (alpha(2A)AR) gene. Because mutation of the alpha(2A)AR gene in the mouse fosters epileptogenesis, we developed methods for analysis of the alpha(2A)AR coding region applicable to any pathophysiologic state in which the alpha(2A)AR could be implicated in the disease mechanism. This study rules out mutations in the alpha(2A)AR coding region as causal for this form of autosomal dominant TLE. PMID- 9855535 TI - Spectrum of disorders associated with enlarged sylvian fissures in infancy. AB - Disproportionate enlargement of the sylvian fissures (ESF) on MRI of an infant's CNS suggests underdevelopment of the cortical operculum. We reviewed charts of infants with isolated ESF. Conditions associated with ESF included feeding difficulties and facial dysmorphism (syndromic or nonsyndromic). There may be an embryologic link between growth of the opercular cortex and development of the face. PMID- 9855536 TI - Pulse cyclophosphamide therapy in chronic inflammatory demyelinating polyneuropathy. AB - Fifteen patients with chronic inflammatory demyelinating neuropathy (CIDP) were treated with pulse intravenous cyclophosphamide (IVCY) monthly for up to 6 months. Eleven patients reached a complete remission; only one patient worsened. Complications included nausea, vomiting, anemia, and hair loss. This case series suggests that monthly IVCY is beneficial in the treatment of CIDP and warrants a controlled study. PMID- 9855537 TI - Molecular mimicry in chronic inflammatory demyelinating polyneuropathy and melanoma. AB - Polyclonal immunoglobulin M antibodies to the monosialoganglioside GM2, sulfoglucuronyl glycolipids, and sulfatide were detected by thin-layer chromatography and enzyme-linked immunosorbent assay in the serum of a patient with melanoma and chronic inflammatory demyelinating polyneuropathy. Both the patient's serum and polyclonal antibodies against GM2 reacted strongly with a biopsy of melanomatous tissue from the patient, suggesting a process of molecular mimicry. PMID- 9855538 TI - Inclusion body myositis: abnormal protein accumulation does not trigger apoptosis. AB - To examine whether apoptosis may contribute to muscle fiber loss in inclusion body myositis, we used the terminal deoxynucleotidyl transferase-mediated X-dUTP nick-end labeling (TUNEL) assay to compare the occurrence of DNA fragmentation in muscle samples from patients with inclusion body myositis and polymyositis. TUNEL positive nuclei in nonnecrotic muscle fibers were rare even in the vicinity of amyloid-like material; significantly more frequent in polymyositis than inclusion body myositis; and several times less frequent than necrotic muscle fibers or mononuclear cell myocytotoxicity in both patient groups. Apoptosis is unlikely to play a significant role in the pathogenesis of inclusion body myositis. PMID- 9855539 TI - The first European family with tibial muscular dystrophy outside the Finnish population. AB - We report the first European tibial muscular dystrophy (TMD) family outside the Finnish population. Clinical examination showed late onset distal leg myopathy similar to the description of TMD. A molecular genetic study was made owing to the very recent TMD linkage findings on chromosome 2q31. All five clinically affected patients segregated a specific haplotype for the locus, whereas two unaffected patients had different haplotype. The results of this family without Finnish ancestors show that TMD exists outside the Finnish population. PMID- 9855540 TI - Molecular basis of hypokalemic myopathy caused by 17alpha-hydroxylase/17,20-lyase deficiency. AB - We describe a 28-year-old woman presenting with hypokalemic myopathy caused by 17alpha-hydroxylase/17,20-lyase deficiency caused by a homozygous mutation consisting of a G-to-C transition in the initiation codon in exon 1 of the CYP17 gene resulting in expression of an enzymatically inactive truncated P450c17 protein. PMID- 9855541 TI - Hereditary spastic paraplegia with a thin corpus callosum and thalamic involvement in Japan. AB - The authors examined two Japanese siblings with a recessive hereditary spastic paraplegia (HSP) with dementia and a thin corpus callosum. Both showed thalamic glucose hypometabolism on PET. Recessive HSP with a thin corpus callosum is a rare disorder, with less than 20 reported patients, that may be a Japanese subtype of HSP. PMID- 9855542 TI - Efficacy of levodopa therapy on motor function after posteroventral pallidotomy for Parkinson's disease. AB - We evaluated motor function in 41 consecutive PD patients undergoing unilateral MRI/microelectrode-guided posteroventral pallidotomy to determine whether the motor effects of levodopa therapy change after surgery. Total Unified Parkinson's Disease Rating Scale motor "off" and motor "on" subscores all improved and the rapidity of action, magnitude, and duration of motor response to levodopa therapy were all maintained after pallidotomy. PMID- 9855543 TI - Mutation, sequence analysis, and association studies of alpha-synuclein in Parkinson's disease. AB - A mutation within the alpha-synuclein gene on human chromosome 4 has been reported to segregate with PD in an Italian family. We screened a sample of familial cases of PD for mutation in the alpha-synuclein gene. None of the familial cases of PD carried a mutation within the alpha-synuclein gene, and no association was detected between PD and alleles of a dinucleotide repeat marker within the alpha-synuclein gene. We conclude that variation within the alpha synuclein gene does not play a significant role in the risk for PD in our sample. PMID- 9855544 TI - High prevalence of neurovascular instability in neurodegenerative dementias. AB - Orthostatic hypotension and carotid sinus hypersensitivity were assessed in patients meeting clinical criteria for dementia with Lewy bodies (DLB; n=30) and AD (n=35). Cardioinhibitory carotid sinus hypersensitivity (CI) was the most common sign (AD patients, 28%; DLB patients, 41%). Preliminary data from a secondary analysis excluding patients with hypertension or EKG evidence of ischemia suggested that CI may be significantly more common in DLB. Larger studies are needed to evaluate the implications for treatment and to explore the underlying mechanisms. PMID- 9855545 TI - Continuous muscle fiber activity associated with morphea (localized scleroderma). PMID- 9855546 TI - Postinfectious pandysautonomia with complete recovery after intravenous immunoglobulin therapy. PMID- 9855547 TI - Refractory status epilepticus: response to ketamine. PMID- 9855548 TI - Multifocal brain MRI hypointensities secondary to embolic metal fragments from a mechanical heart valve prosthesis: a possible source of epileptic seizures. PMID- 9855549 TI - Giant cell arteritis presenting as cluster headache. PMID- 9855550 TI - Nonpositional headache caused by spontaneous intracranial hypotension. PMID- 9855551 TI - Granulomatous compressive thoracic myelopathy as the initial manifestation of Wegener's granulomatosis. PMID- 9855552 TI - Impaired motor cortex inhibition in patients with ALS: evidence from paired transcranial magnetic stimulation. PMID- 9855553 TI - Neurology's witness to history: Leo Alexander. PMID- 9855554 TI - Autosomal recessive posterior column ataxia and retinitis pigmentosa. PMID- 9855555 TI - Dissociation of spinothalamic sensation in lateral medullary infarction. PMID- 9855556 TI - Idiopathic granulomatous angiitis of the CNS manifesting as diffuse white matter disease. PMID- 9855557 TI - Idiopathic granulomatous angiitis of the CNS manifesting as diffuse white matter disease. PMID- 9855558 TI - Acute myopathy after liver transplantation. PMID- 9855559 TI - Antiparkinsonian and anti-levodopa-induced dyskinesia effects obtained by stimulating the same site within the GPi in PD. PMID- 9855560 TI - Cerebellar input tremor: inferior or superior cerebellar peduncle lesion? PMID- 9855561 TI - Systematic reviews. Cochrane Stroke Group. PMID- 9855562 TI - Vigabatrin-associated retinal cone system dysfunction. PMID- 9855563 TI - Vigabatrin-associated retinal cone system dysfunction. PMID- 9855564 TI - Vigabatrin-associated retinal cone system dysfunction. PMID- 9855565 TI - Remember the Little Red Hen? PMID- 9855566 TI - A review of plants used in the treatment of liver disease: part 1. AB - Botanicals have been used traditionally by herbalists and indigenous healers worldwide for the prevention and treatment of liver disease. Clinical research in this century has confirmed the efficacy of several plants in the treatment of liver disease. Basic scientific research has uncovered the mechanisms by which some plants afford their therapeutic effects. Silybum marianum (milk thistle) has been shown to have clinical applications in the treatment of toxic hepatitis, fatty liver, cirrhosis, ischemic injury, radiation toxicity, and viral hepatitis via its antioxidative, anti-lipid peroxidative, antifibrotic, anti-inflammatory, immunomodulating, and liver regenerating effects. Picrorhiza kurroa, though less well researched than Silybum, appears to have similar applications and mechanisms of action. When compared with Silybum, the hepatoprotective effect of Picrorhiza was found to be similar, or in many cases, superior to the effect of Silybum. PMID- 9855567 TI - Botanical influences on cardiovascular disease. AB - Several botanicals, including Crataegus oxycantha, Terminalia arjuna, Inula racemosa, and Astragalus membranaceus, have been found to have therapeutic benefit for the treatment of cardiovascular disease. Crataegus oxycantha has been used traditionally as a cardiac tonic and current uses include treatment for angina, hypertension, arrhythmias, and congestive heart failure. Animal studies have also indicated that Crataegus extracts may also have potential use as anti ischemic and lipid-lowering agents. The bark of the Terminalia arjuna tree has a long history of use as a cardiac tonic as well, and has been indicated in the treatment of coronary artery disease, heart failure, hypercholesterolemia and for relief of anginal pain. Additionally, it has been found to have antibacterial and antimutagenic properties. Inula racemosa, also known as Pushkarmoola, is another traditional Ayurvedic botanical that has potential cardioprotective benefit. In human trials, a combination of Inula racemosa and Commiphora mukul was shown to be superior to nitroglycerin in reducing the chest pain and dyspnea associated with angina. Astragalus membranaceus, a Chinese herb, is often used as a "Qi tonifier" and has been studied for its therapeutic benefit in treatment of ischemic heart disease, myocardial infarction, heart failure, and relief of anginal pain. Clinical studies have indicated that its in vitro antioxidant activity is the mechanism by which it affords its cardioprotective benefit. PMID- 9855568 TI - Inositol--clinical applications for exogenous use. AB - Recent advances in nutritional and biochemical research have documented inositol as an important dietary and cellular constituent. The processes involved in inositol metabolism and its derivatives in the tissues of mammals have been characterized in vivo as well as at the enzymatic level. Biochemical functions defined for phosphatidylinositol in biological membranes include the regulation of cellular responses to external stimuli and/or nerve transmission as well as the mediation of enzyme activity through interactions with various specific proteins. Altered production of inositol has been documented in patients with diabetes mellitus, chronic renal failure, galactosemia, and multiple sclerosis. Inositol has been reported to be effective in treating central nervous system disorders such as depression, Alzheimer's disease, panic disorder, and obsessive compulsive disorder. It has documented benefit for use in pediatric respiratory depression syndrome. In addition, recent studies have evaluated its usefulness as an analgesic. Inositol has been studied extensively as potential treatment to alleviate some negative effects associated with lithium therapy. The use of inositol in pregnant women remains controversial. Although its benefit in preventing neural tube defects in embryonic mice is documented, the risk of inducing uterine contractions limits its usefulness in pregnancy. PMID- 9855569 TI - Perillyl alcohol: applications in oncology. AB - Perillyl alcohol is a monoterpene isolated from the essential oils of lavendin, peppermint, spearmint, cherries, celery seeds, and several other plants. In animal studies it has been shown to regress pancreatic, mammary, and liver tumors, to exhibit possible application as a chemopreventative agent for colon, skin, and lung cancer, and as a chemotherapeutic agent for neuroblastoma, and prostate and colon cancer. Perillyl alcohol is active in inducing apoptosis in tumor cells without affecting normal cells and can revert tumor cells back to a differentiated state. Its mechanism of action is unclear, but it has actions on various cellular substances which control cell growth and differentiation. It has been shown to increase mannose-6-phosphate/insulin-like growth factor II receptors, increase tissue growth factor beta receptors, increase Bak, decrease ras protein prenylation, decrease ubiquinone synthesis, and induce Phase I and Phase II detoxification systems. Preliminary human trials have not demonstrated tumor regression at a four times daily dosage schedule. In addition, significant side-effects, mainly gastrointestinal, have been experienced. PMID- 9855571 TI - Methylcobalamin. AB - Methylcobalamin is one of the two coenzyme forms of vitamin B12. Evidence indicates this form of vitamin B12, in addition to having a theoretical advantage over cyanocobalamin, actually has some metabolic and therapeutic applications not shared by the other forms of vitamin B12. This monograph provides an overview of the pharmacokinetics of methylcobalamin, and will highlight the potential therapeutic relevance for Bell's palsy, cancer, diabetic neuropathy, eye function, heart rate variability, HIV, homocysteinemia, male impotence, and sleep disorders. PMID- 9855570 TI - Piper methysticum (kava kava). AB - Piper methysticum (kava kava) is a plant native to the Pacific Island region, and has been used ceremonial for thousands of years. The active ingredients are a group of substances know as kava lactones (AKA kava pyrones). Four lactones in kava have been found to have significant analgesic and anesthetic effects via non opiate pathways. Kava's most popular application is as a natural anxiolytic, comparing favorably in several studies to a number prescription medications, including benzodiazepines. CNS effects seem to be mediated by several mechanisms. Studies have been conflicting regarding its GABA-receptor-binding capacity, although this has been found to occur in some studies. In vitro kava has been found to block norepinephrine uptake. It also has some anti-convulsant capabilities, which appear to be mediated by Na+ channel receptor sites. The therapeutic dosage is in the range of 50-70 mg kava lactones three times daily. The most common side effect, usually seen only with long-term, heavy usage of the herb, is a scaly skin rash called "kava dermopathy." It has also been know to potentiate other medications such as barbiturates and Xanax. PMID- 9855572 TI - Envelopes: presidential address. AB - The fabric of medical care has been altered during the past 2 decades. Initially this alteration was furtive but it rapidly escalated, producing change that will be irrevocable. Physicians, dedicated to caring for their patients, frequently were found struggling in the wake of the changes in the health care system. In most situations they found themselves swept aside from crucial decision making because of timidity, practice obligations, and absence of knowledge related to "medical business." With about 600 managed care operations in the United States, physicians can no longer afford to sit in the stands and just watch the play on the field. Participation is crucial and must be accompanied by additional learning, such as masters degrees in business administration or MBAs. This will allow the new players to read the playbook of business medicine. Although on-the job training is possible, it usually has a slow learning curve as a result of diluting the new assignment with practice obligations. Despite these formidable challenges, physicians must enter the local, state, and national arenas and participate. Despite change and reaction to change, physicians have an irrevocable trust that cannot be withdrawn or overlooked. It has been earned and pervades every facet of our professional careers. The physician has an uncompromising duty and privilege to care for his or her patient in the highest and most ethical manner, a duty that will remain forever. This will never change! PMID- 9855573 TI - Comparison of ureteral and cervical descents during vaginal hysterectomy for uterine prolapse. AB - OBJECTIVE: The study measured ureteral and cervical locations during vaginal hysterectomy for prolapse and the extent of parametrial ligament shortening possible. STUDY DESIGN: Cervical and ureteral position were measured in 26 women undergoing uterine prolapse correction. Parametrial clamp tip location was also measured. RESULTS: The cervix lay between 0 and -14.5 cm (below) the hymen (mean +/- SD -5.35 +/- 3.96 cm) and the ureters lay +5.0 to -4.0 cm (mean +/- SD +1.89 +/- 1.99 cm). Correlation of ureteral with cervical position was 0.69 (P <.01) and correlation with ipsilateral uterosacral ligament clamp positions was 0.80 (P <.01). Regression line slope relating cervical descent and cervix to ureter distance was 0.65, indicating that for every 3 cm of cervical descent there was 2 cm widening of the gap between the cervix and ureters and 1 cm descent of the ureter. CONCLUSION: For every 3 cm of cervical descent the ureters descend 1 cm, thereby widening the ureterocervical gap and permitting ligament shortening during vaginal hysterectomy. PMID- 9855574 TI - The Noble-Mengert-Fish operation-revisited: a composite approach for persistent rectovaginal fistulas and complex perineal defects. AB - OBJECTIVE: Our aims were to evaluate the full-thickness anterior rectal wall advancement flap in the treatment of primary and recurrent or persistent rectovaginal fistulas, evaluate the surgical exposure for composite repair of site-specific perineal defects, and categorize clinical manifestations of site specific perineal defects caused by obstetric injury. STUDY DESIGN: This is a prospective study of all patients with fecal incontinence from rectovaginal septal defects and complex perineal obstetric injuries treated by the Noble Mengert-Fish operation. RESULTS: Thirty-four patients were classified into groups on the basis of site-specific perineal defects. Anatomic success was 94.2%. Functional success was excellent in 76.5%, good in 14.7%, fair in 5.9%, and poor in 2.9%. CONCLUSION: The Noble-Mengert-Fish operation is effective for primary and recurrent or persistent rectovaginal fistulas. The circumanal surgical exposure permits concomitant repair of all perineal defects. PMID- 9855575 TI - Site-specific fascial defects in the diagnosis and surgical management of enterocele. AB - OBJECTIVE: The aim of this study was to assess the surgical feasibility and clinical outcomes of a vaginal enterocele repair that was based on the theory of site-specific defects in the vaginal fascia. STUDY DESIGN: Seventeen patients during a 2-year period with a diagnosis of enterocele and vaginal vault descensus with or without coexisting rectocele underwent surgical correction with a site specific fascial defect repair. An enterocele was defined as vaginal wall prolapse seen during the operation in which the peritoneum was found to be in direct contact with the vaginal epithelium, with no intervening fascia. Patients were examined at 4 weeks after the operation and then at 6-month intervals, with site-specific analysis of pelvic prolapse at the vaginal apex and posterior vaginal segment. RESULTS: Identification and site-specific fascial defect repair of the enterocele were successfully performed in all 17 cases. All patients also underwent a uterosacral ligament vaginal vault suspension, and 15 patients (88%) underwent concurrent posterior colporrhaphy. There were no intraoperative complications. At a mean follow-up of 6.3 months (range 1-17 months), 2 patients (12%) had mild, asymptomatic vaginal vault descensus but no patients (0/17) had evidence of a recurrent enterocele or rectocele. CONCLUSION: Enterocele correction through a fascial defect repair is easily performed through the vaginal route and is associated with excellent surgical outcomes on short-term follow-up. PMID- 9855576 TI - Conservative surgical management of Mersilene mesh suburethral sling erosion. AB - OBJECTIVE: The aim of this study was to describe conservative surgical management of polyethylene terephthalate (Mersilene, Ethicon, Inc, Somerville, NJ) mesh suburethral sling erosion as an alternative to sling removal. STUDY DESIGN: Seven women who had undergone suburethral sling procedures with Mersilene mesh were found to have varying degrees of mesh visible in the vagina at 4 to 12 weeks after the operation. Three women with erosions <5 mm were treated in the office and 4 women who had 6 to 30 mm erosions underwent inpatient surgical revision, which consisted of trimming of mesh when needed, excision of granulation tissue, and re-covering of the erosion in 2 layers. RESULTS: All 7 patients have been followed up for 6 to 37 months. Only 1 woman who underwent inpatient surgical revision had a recurrence (4 mm); this was treated successfully in the office. CONCLUSION: Vaginal erosions in suburethral sling procedures with Mersilene mesh can be managed initially with our conservative surgical technique. PMID- 9855577 TI - Synthetic graft placement in the treatment of fascial dehiscence with necrosis and infection. AB - OBJECTIVE: The objective of this study was to describe the use of synthetic grafts in repairing fascial dehiscence complicated by fascial necrosis and infection after obstetric and gynecologic operations. STUDY DESIGN: A retrospective review of the operating room records at Hutzel Hospital (Detroit, Mich) was performed to find all cases of fascial dehiscence repaired during a 6 year period between January 1, 1991, and December 31, 1996. Patients with partial or complete disruption of the fascia with evidence of fascial necrosis and infection were included in this study. Demographic information; the initial surgical procedure, including type of incision; suture material; use of synthetic graft and closure technique for repair of dehiscence; postoperative complications, microbiologic results; antibiotic therapy; subsequent operations; length of hospital stay; and late complications were recorded. RESULTS: During the study period 52 patients underwent repair of fascial dehiscence; 36 of these had concurrent fascial necrosis and infection, including 4 women with necrotizing fasciitis. Eighteen patients were from the obstetric service and 18 were from the benign or cancer gynecology service. Ninety-one bacterial isolates were recovered from the infected wounds. Extensive fascial resection precluded closure without tension in 18 cases and necessitated synthetic graft placement to prevent evisceration. Graft materials included polypropylene (11 cases) and polyglactin (7 cases). Late complications of graft placement included extrusion of the graft in 3 patients and incisional hernia in 1. CONCLUSIONS: Extensive fascial debridement with resection prevents primary closure of wound dehiscence. Synthetic grafts permit primary closure of large fascial defects and can be used with extensive debridement in the presence of infection. PMID- 9855578 TI - Paravaginal repair of lateral vaginal wall defects by fixation to the ischial periosteum and obturator membrane. AB - OBJECTIVE: The aim of the study was to evaluate the anatomic basis, efficacy, and safety of a technique for correcting lateral wall vaginal defects. STUDY DESIGN: Phase I was cadaveric dissection carried out to ascertain the strength and position of structures likely to support lateral vaginal wall defects. The ischial periosteum just anterior to the ischial spine was found to be strong tissue, relatively free of nerves and vessels. In phase II, paravaginal defects were repaired by placing sutures through the arcus tendineus and underlying obturator fascia, obturator membrane, and ischial periosteum. Other defects and urinary incontinence were corrected within the same surgical setting. Forty patients were followed up for an average of 39 months (range 7-52 months). Preoperative evaluation consisted of an extensive history, cough stress test, spontaneous uroflowmetry, postvoid residual urine determination, urethral axis determination, site-specific pelvic floor defect evaluation, and multichannel urodynamic studies. After the operation patients underwent evaluations at 3 months, at 6 months, and then annually. RESULTS: Objective site-specific re examination of the 40 patients revealed the following recurrences: lateral wall in 1 of 40 procedures, anterior wall in 3 of 35 procedures, posterior wall in 1 of 36 procedures, and apical wall in 1 of 27 procedures. Thirty-four of 36 women (94.4%) with urodynamically confirmed genuine stress incontinence or potential incontinence achieved cure (P <.001). CONCLUSIONS: (1) The ischial periosteum and obturator membrane are consistently strong reattachment sites. (2) Repair of paravaginal defects with these tissues is effective and safe. (3) Urodynamic parameters were unchanged after the operation except for measures of incontinence, which were improved (P <.001). (4) Performing other pelvic procedures did not negatively alter the success rates of paravaginal repair. (5) The urethral axis was favorably altered after the operation (P <.01). PMID- 9855579 TI - Posterior vaginal prolapse and bowel function. AB - OBJECTIVE: This study's objectives were to describe symptoms related to bowel dysfunction in women with uterovaginal prolapse and to compare these symptoms according to extent of posterior vaginal prolapse. STUDY DESIGN: One hundred forty-three women completed a questionnaire assessment of bowel function and underwent standardized physical examination according to the International Continence Society's system for grading uterovaginal prolapse. RESULTS: The mean age was 59.2 years (SD 11.8 years); 78% of the women were postmenopausal. According to the furthest extent of posterior vaginal prolapse at point Bp, 22 (15.5%) were in stage 0, 46 (32.4%) were in stage I, 50 (35.2%) were in stage II, 23 (16.2%) were in stage III, and 1 (0.7%) was in stage IV. Ninety-two percent of women reported having bowel movements at least every other day. When asked whether straining was required for them to have a bowel movement, 38 (26.6%) reported never or rarely, 71 (49.6%) reported sometimes, 20 (14.0%) reported usually, and 14 (9.8%) reported always. When asked whether they ever needed to help stool come out by pushing with a finger in the vagina or rectum, 98 (69.0%) reported never or rarely, 30 (21.1%) reported sometimes, 8 (5.6%) reported usually, and 6 (4.2%) reported always. Twenty-three women (16.1%) had fecal incontinence, with 11 having loss of control of stool less often than once a month and 12 having it more often than once a month. When asked whether to rate how much they were bothered by their bowel function on a scale of 1 to 10, with 1 being not at all and 10 being extremely, 51.7% of women chose 1 to 4, 20.3% chose 5 to 7, and 28% chose >/=8. There were no clinically significant associations between any of the questions related to bowel function and severity of posterior vaginal prolapse. CONCLUSION: Women with uterovaginal prolapse frequently have symptoms related to bowel dysfunction, but this is not associated with the severity of posterior vaginal prolapse. PMID- 9855580 TI - An anatomic and functional assessment of the discrete defect rectocele repair. AB - OBJECTIVE: The aim of this study was to describe the anatomic and functional results of the discrete fascial defect rectocele repair. STUDY DESIGN: Sixty-nine women underwent rectocele repair at Duke University Medical Center during a 3 year period beginning January 1, 1994. Repair was limited to reapproximation of discrete defects in the rectovaginal fascia, without levator plication or perineorrhaphy. Outcome measures included Pelvic Organ Prolapse Quantitation measurements, prolapse stage, and a symptom questionnaire. Univariate and nonparametric tests were used as appropriate. RESULTS: Before the operation 46% patients (32/69) reported constipation, 39% (27/69) reported splinting, 32% (22/69) reported tenesmus, and 13% (9/69) reported fecal incontinence. The median preoperative posterior Pelvic Organ Prolapse Quantitation stage was 2 (1-4). Pelvic Organ Prolapse Quantitation stage had improved for all but 2 women at 6 weeks. Eighteen percent (8/43) had recurrent rectoceles at 12 months. Mean values for the points describing the posterior vaginal wall improved >2 cm (P <.0001). Although perineorrhaphy was not performed, the genital hiatus decreased by 2. 3 cm (P <.0001), with no significant change in the length of the perineal body. Functional results mirrored anatomic results, with statistically significant improvements for all symptoms. CONCLUSIONS: The discrete defect rectocele repair provides anatomic correction of rectoceles with alleviation of associated symptoms for most women. PMID- 9855581 TI - Teaching the pelvic organ prolapse quantitation system. AB - OBJECTIVE: The objective of this study was to determine the ease with which the new Pelvic Organ Prolapse Quantitation system could be learned by residents and medical students. STUDY DESIGN: Standardized multiple-choice tests were administered to 51 obstetric and gynecology residents and medical students from 4 community-based and university-based programs. Parallel pretests and posttests were administered in conjunction with a 17-minute video demonstration of the Pelvic Organ Prolapse Quantitation system and with the addition of a visual memory aid. The posttest was repeated 3 months after the video presentation. RESULTS: The use of a 17-minute video significantly enhanced participants' ability to interpret examination findings when expressed in the terminology of the system (mean improvement in scores 33%, P <.0001). Posttest scores were similar regardless of the type of program or exposure to urogynecology faculty. These scores were maintained at the 3-month retesting. CONCLUSION: The Pelvic Organ Prolapse Quantitation system can be effectively taught by means of a public domain video presentation. PMID- 9855582 TI - The risk of cystocele after sacrospinous ligament fixation. AB - OBJECTIVE: The aim of this study was to determine whether sacrospinous ligament fixation independently increases the risk of anterior vaginal wall defect. STUDY DESIGN: A retrospective cohort study was conducted on patients undergoing pelvic reconstructive surgical operations by a single surgeon from 1970 through 1997. Two groups were examined and divided into subgroups to evaluate the effects of exposure to sacrospinous ligament fixation: patients with anterior wall defects undergoing standard anterior colporrhaphy with (group 1A) or without (group 1B) concomitant sacrospinous ligament fixation, and patients without anterior wall defects undergoing other pelvic reconstructive procedures (but not anterior colporrhaphy) with (group 2A) or without (group 2B) sacrospinous ligament fixation. Recurrence rates were calculated for each group according to evidence of any degree of anatomic defect. RESULTS: Among 322 patients in group 1, 9 of 77 in group 1A and 23 of 245 in group 1B (11.7% vs 9.4%, P >.05) had anterior wall recurrences. Among 73 patients in group 2, 8 of 45 in group 2A and 5 of 28 in group 2B (17.8% vs 17.9%, P >.05) had subsequent anterior wall defects. CONCLUSION: The occurrence of anterior vaginal wall defects was not found to be altered by the performance of sacrospinous ligament fixation. These findings may be attributable to surgical technique emphasizing maintenance of anterior vaginal wall length during sacrospinous ligament fixation. PMID- 9855583 TI - Vaginal hysterectomy in women with history of previous cesarean delivery. AB - OBJECTIVE: This study aimed to compare surgical outcomes with vaginal hysterectomy between women who have had >/=1 cesarean delivery and those who have not had a cesarean delivery. A secondary objective was to analyze the effect of previous vaginal birth on the complication rate of vaginal hysterectomy. STUDY DESIGN: A retrospective analysis was performed on 221 women undergoing vaginal hysterectomy. Women were separated into those who had a history of previous cesarean deliveries (N = 35) and those who did not (N = 186). The groups were analyzed for the indications for surgery, perioperative hemoglobin loss, operative time, length of hospitalization, and complications. Trends in the complication rate for women in the previous cesarean group were also studied from the perspectives of numbers of previous cesarean deliveries and of vaginal delivery history. The 95% confidence intervals for the difference between proportions as well as P values for probability tests were calculated. P <.05 was considered significant. RESULTS: Previous cesarean delivery experience did not affect hemoglobin change, hospital stay, or operative time among women undergoing vaginal hysterectomy. A total of 11.3% of women in the previous cesarean group had complications, versus 4.3% for the noncesarean group (P =.10, 95% confidence interval -3.8% to 18.0%). Complications did not increase with increasing number of previous cesarean deliveries (U = 1020.5, P =.28). Also, a trend toward fewer complications among patients with a history of cesarean delivery who had also had a vaginal delivery was demonstrated (U = 836, P =.05). Overall, women who were undergoing vaginal hysterectomy who had a history of >/=1 previous vaginal delivery had a complication rate of 3.2%, versus 17.6% for women who had not had a previous vaginal birth (P =.004, 95% confidence interval -27.5% to -1.3%). CONCLUSIONS: In this study women who had a history of previous cesarean delivery were not at higher risk for greater hemoglobin loss, longer hospital stay, more prolonged operative time, or significantly more perioperative complications when undergoing vaginal hysterectomy than were those women who had no history of previous cesarean delivery. Likewise, increasing the number of previous cesarean deliveries did not have an adverse impact on the complication rate. Previous vaginal delivery lowered the risk of complications from vaginal hysterectomy. PMID- 9855584 TI - Selective estrogen receptor modulators: a new category of therapeutic agents for extending the health of postmenopausal women. AB - Selective estrogen receptor modulators are a new category of therapeutic agents that bind with high affinity to estrogen receptors and mimic the effect of estrogens in some tissues but act as estrogen antagonists in others. Tamoxifen, a triphenylethylene derivative, was the first clinically available selective estrogen receptor modulator. It is a potent antiestrogen in the breast, and its use in breast cancer patients has made it the most prescribed antineoplastic drug worldwide. It has estrogen-like activity on bone metabolism, and it also reduces cholesterol. However, its ability to produce proliferation, polyp formation, and even carcinomas in the endometrium is well known. A new selective estrogen receptor modulator, raloxifene, a benzothiopene derivative, has a clinical profile similar to that of tamoxifen. However, both preclinical and clinical studies reveal that, unlike tamoxifen, it is a pure antiestrogen in the uterus. It has recently been approved by the Food and Drug Administration for prevention of osteoporosis in postmenopausal women. This report reviews pertinent preclinical and currently available clinical studies about this new selective estrogen receptor modulator and discusses clinical applicability. PMID- 9855585 TI - Treatment for lactation suppression: little progress in one hundred years. AB - Our goal was to characterize the postpartum symptoms experienced by women who do not breast-feed and to review data on the efficacy of nonpharmacologic methods of lactation suppression. The placebo arms of randomized clinical trials of pharmacologic methods for lactation suppression were used to characterize postpartum symptoms. A subset of the placebo arms was reviewed to assess current strategies for treatment of symptoms associated with lactation suppression. Studies of nonpharmacologic methods of lactation suppression were also reviewed to assess efficacy. Engorgement and breast pain may encompass most of the first postpartum week. Up to one third of women who do not breast-feed and who use a brassiere or binder, ice packs, or analgesics may experience severe breast pain. Specific studies of nonpharmacologic methods of lactation suppression were limited and inconclusive. Available data suggest that many women using currently recommended strategies for treatment of symptoms may nevertheless experience engorgement or pain for most of the first postpartum week. PMID- 9855586 TI - Pregnancies after radical vaginal trachelectomy for early-stage cervical cancer. AB - OBJECTIVE: The purpose of this study was to evaluate the role of fertility preserving surgery in the treatment of early-stage cervical cancer. STUDY DESIGN: We retrospectively reviewed our first 30 patients treated by laparoscopic pelvic lymphadenectomy, followed by radical vaginal trachelectomy, from October 1991 to April 1998. RESULTS: The median age of the patients was 32 years (range 22-42 years); 15 were nulligravid and 19 nulliparous. Twenty cancers were at stage IB, 1 was at stage IA1, 7 were at stage IA2, and 2 were at stage IIA. The majority (18/30) were squamous. Two lesions were >2 cm in size, and only 4 had vascular space invasion. The median operative time was 285 minutes (range 155-455 minutes), median blood loss 200 mL (range 50-1200 mL), and median hospital stay 4 days (range 2-9 days). There were 4 intraoperative complications-2 attributed to the trachelectomy and 2 resulting from the lymphadenectomy. The current median follow-up time is 25 months (range 1-79 months). One patient had a recurrence in the left parametrium 18 months after vaginal radical trachelectomy and died of metastatic disease. The only 6 patients attempting pregnancy so far have succeeded: 4 have had healthy babies delivered by cesarean section at 39, 38, 34, and 25 weeks of gestation. Two are currently 33 and 8 weeks pregnant. CONCLUSION: Radical vaginal trachelectomy appears to be a valuable procedure in well-selected patients with early-stage cervical cancer. Successful pregnancies are definitely possible after this procedure. This new surgical technique warrants further careful evaluation to determine precise indications. PMID- 9855587 TI - A population-based study of human papillomavirus deoxyribonucleic acid testing for predicting cervical intraepithelial neoplasia. AB - OBJECTIVE: Our purpose was to determine the predictive values of primary or secondary screening for cervical human papillomavirus infection for cytologic detection of cervical intraepithelial neoplasia. STUDY DESIGN: Most of the 254 women referred for colposcopy in Vasterbotten County in Sweden during October 1993 through December 1995 and 320 age-matched women from the general population were screened for human papillomavirus deoxyribonucleic acid by nested general primer polymerase chain reaction. RESULTS: Ninety-six percent of women with high grade cervical intraepithelial neoplasia had human papillomavirus, compared with 4% of women with normal findings (odds ratio 606; 95% confidence interval 137 to 5607). Thirty-seven percent of referred women and 48% of referred women >39 years old had mostly minor cytologic abnormalities with no human papillomavirus. The human papillomavirus-associated positive predictive value for cervical intraepithelial neoplasia was 76% in the colposcopy group and 11% in the general population, whereas the negative predictive value was >97% in both populations. CONCLUSION: Testing for human papillomavirus deoxyribonucleic acid seems diagnostically useful among women referred for colposcopy. PMID- 9855588 TI - The effect of a high partial pressure of carbon dioxide environment on metabolism and immune functions of human peritoneal cells-relevance to carbon dioxide pneumoperitoneum. AB - OBJECTIVE: Our purpose was to evaluate in vitro the effect of a high partial pressure of carbon dioxide environment used in laparoscopy on metabolic and immune response of various human peritoneal cells. STUDY DESIGN: Polymorphonuclear leukocytes were obtained from 5 healthy volunteers, peritoneal macrophages were obtained from the effluent of 8 patients undergoing continuous ambulatory peritoneal dialysis, and human peritoneal mesothelial cell cultures were prepared from omentum derived from 5 patients undergoing elective surgery. The cells were exposed to a laparoscopy-like environment (1 atmosphere carbon dioxide and 0.2 atmosphere oxygen), to a control gas mixture (1 atmosphere helium and 0.2 atmosphere oxygen), or air for 3 hours. After exposure to gas mixtures, cell functions were tested at various recovery periods. RESULTS: Three hours of exposure to a high partial pressure of carbon dioxide had no effect on viability of peritoneal macrophages and human peritoneal mesothelial cells, tested by trypan blue dye uptake and lactate dehydrogenase release. A high partial pressure of carbon dioxide decreased the mitochondrial dehydrogenases activity of peritoneal macrophages and human peritoneal macrophage cells by 60%, assayed by 3 (4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction. High partial pressure of carbon dioxide blocked the superoxide release from activated polymorphonuclear leukocytes and the secretion of interleukin 1beta from stimulated peritoneal macrophages, and human peritoneal macrophage cells were decreased by 15% and 30% and the secretion of tumor necrosis factor-alpha from peritoneal macrophages was suppressed by 85%. Mitochondrial activity, polymorphonuclear leukocyte function, and interleukin 1beta and tumor necrosis factor-alpha secretion returned to normal after a recovery period of 12 to 24 hours, 4.5 hours, and 24 hours, respectively. In the control experiments exposure of cells to helium had no suppressive effect. CONCLUSIONS: Exposure of cells to a high partial pressure of carbon dioxide environment suppresses the inflammatory and metabolic responses of peritoneal cells. We suggest that this suppressive effect may contribute to the low postsurgery adhesion formation and the reduction in postoperative pain observed in laparoscopy. Nevertheless, the suppression of the immune response should also be taken into account for operations involving a high risk of bacterial dissemination. PMID- 9855589 TI - Role of fascial collagen in stress urinary incontinence. AB - OBJECTIVES: Our purpose was to determine whether collagen of the pubocervical fasciae that support the urethrovesical junction undergoes alterations that might contribute to incontinence. STUDY DESIGN: Pubocervical fascia was collected as a residual tissue in 82 patients, aged 25 to 73 years, during surgical treatment of cystocele (n = 26, no incontinence) or of stress urinary incontinence (n = 56). Measurements were made of collagen content, solubility, and cross-linking and of collagenase activity. RESULTS: Patients treated for incontinence had the same mean age and parity as the control cystocele group. There was a highly significant (20%, P <.0005) decrease in collagen content in fascial tissue from incontinent women. There was no difference in the percentage of acid-soluble (0.7%) and pepsin-soluble (17%) collagen in the 2 groups of patients; this agrees with the lack of significant change in the degree of collagen cross-linking by pyridinoline. Collagenase activity was significant in fascia but did not change in incontinence. Incontinent women had an increased body mass index. CONCLUSIONS: The pubocervical fasciae of incontinent women show a diminished content of collagen, but this is not accompanied by changes in collagen solubility or cross linking or in collagenase activity. This decrease in collagen may contribute to the weakening of support of the bladder neck. PMID- 9855590 TI - The initiation, continuation, and termination of spontaneous episodes of circus movements in the pregnant myometrium of the rat. AB - OBJECTIVES: The objectives of this study were to determine the mechanisms responsible for the initiation and the termination of episodes of circus movements and to describe the characteristics of these circuits in rat myometrium. STUDY DESIGN: By means of simultaneous recordings from 240 sites in Wistar rats at 17, 19, or 21 days' gestation, a spatial and temporal analysis was performed on the electrical activation from the beginning until the end of 22 episodes of circus movements. RESULTS: The initiation of reentry was caused by (1) transient unexcitable areas at the beginning of the burst or (2) lines of conduction blocks. Circus movements were terminated by (1) reversal of recruitment (decruitment) at the end of the burst, (2) occurrence of lines of conduction blocks, or (3) interruption of the circuit. On average circuits rotated for 6 revolutions during 2 seconds. In most cases circuits remained stable; sometimes circuits drifted along the tissue and occasionally multiple wavelets were seen instead of a single circuit. CONCLUSIONS: Spatial nonhomogeneities in conduction caused by transient unexcitable areas play a major role in both the initiation and the termination of episodes of circus movements in the pregnant rat myo-metrium. In addition, most circuits remained in a single location, although sometimes meandering or even fibrillatory activity was seen. PMID- 9855591 TI - The influence of epidural analgesia on cesarean delivery rates: a randomized, prospective clinical trial. AB - OBJECTIVE: The effects of epidural analgesia on the progress of labor are controversial. The objective of this study was to determine the effect of epidural analgesia on cesarean delivery rates in a population of patients randomly assigned to receive either epidural analgesia or intravenous opioids for intrapartum pain relief. STUDY DESIGN: From January 1995 to May 1996, 318 spontaneously laboring, term, nulliparous patients were randomly assigned to receive either intravenous opioids or epidural analgesia for pain relief. Labor was managed according to the principles of active management of labor. Cesarean delivery was performed for obstetric indications. Data analysis was conducted on an intent-to-treat basis. A subanalysis was subsequently performed on patients who were compliant with the allocated form of treatment. RESULTS: One hundred sixty-two patients were randomly assigned to receive intravenous meperidine and 156 were randomly assigned to receive epidural analgesia. Maternal age, gravidity, race, gestational age, and cervical dilatation at admission and at first analgesic dose did not differ between the groups. Intent-to-treat data analysis revealed no significant difference in the cesarean delivery rate between the 2 groups, being 13.6% in the opioid group and 9.6% in the epidural group (relative risk 0.70, 95% confidence interval 0.38-1.31, P >.05). Cesarean delivery rates for the indication of dystocia also did not differ, being 10.5% in the opioid group and 5.8% in the epidural group (relative risk 0.56, 95% confidence interval 0.26-1.21, P >.05). Subanalysis of the data from patients who were compliant with the allocated form of treatment revealed that patients in the epidural group (n = 147) were 3 times more likely to have an active phase duration >/=8 hours and were 10 times more likely to require >/=2 hours in the second stage of labor than were those in the opioid group (n = 78). There were no significant differences in cesarean delivery rates in this subanalysis, being 7.7% in the opioid group and 8.8% in the epidural group (relative risk 1.15, 95% confidence interval 0.45-2.91, P >. 05). The cesarean delivery rates for dystocia were also similar in the subanalysis, being 3.8% in the opioid group and 5.5% in the epidural group (relative risk 1.42, 95% confidence interval 0.39-5. 22, P >.05). CONCLUSION: Epidural analgesia provides safe and effective intrapartum pain control and may be administered without undesirable effects on labor outcome. PMID- 9855592 TI - Vascular smooth muscle oxygen consumption is reversibly stimulated by sera from women with preeclampsia. AB - OBJECTIVE: Preeclampsia is a complication of pregnancy that causes maternal vasoconstriction and hypertension. The disease may progress to eclampsia, which is thought to be related to cerebral vasospasm. Although there is evidence for more than one circulating factor that causes endothelial cell dysfunction in preeclampsia, little work has focused on the possibility that vascular smooth muscle function might be directly stimulated by a circulating factor. The aim of this study was to determine whether such a factor or factors could be detected by the vessels. STUDY DESIGN: Excessive vascular smooth muscle oxygen consumption was used as a screen for metabolic stimulation because pathologic arterial constriction would require oxidative metabolism to generate adenosine triphosphate. De-endothelialized porcine carotid artery (a well-validated model of human arterial contractile function) was exposed to sera from patients with preeclampsia (1:30 dilution) in a sealed chamber with an oxygen electrode, and the rate of oxygen consumption by the tissue was measured. Comparisons with the effects of sera from matched normal pregnant patients and from nonpregnant women were made. RESULTS: Exposure of vascular smooth muscle to sera from women with preeclampsia for 90 minutes resulted in greater oxygen consumption by the tissue (0.66 +/- 0.16 micromol O2 /min per gram of dry weight) than did exposure to sera of matched pregnant and nonpregnant control subjects (0.34 +/- 0.08 micromol O2 /min per gram of dry weight, P <.001, and 0.29 +/- 0.03 micromol O2 /min per gram of dry weight, P <.001, respectively). This stimulation was completely reversed by rinsing. CONCLUSIONS: There is a factor in the circulation of women with preeclampsia that has the reversible effect on vascular smooth muscle of accelerating oxygen consumption. We discuss the implications of this observation in terms of known aspects of vascular smooth muscle contractile function. PMID- 9855593 TI - Preeclampsia is associated with reduced serum levels of placenta growth factor. AB - OBJECTIVES: Adequate vascular development of the placental bed is essential for normal pregnancy. We assessed serum levels of placenta growth factor, an angiogenic factor, throughout normal pregnancy and determined its association with preeclampsia. STUDY DESIGN: Serum samples were collected from (1) 308 healthy pregnant women throughout normal gestation, (2) at delivery from 30 each gestational age-matched patients with normal pregnancy and preeclampsia, and (3) maternal and cord blood samples from normal deliveries with and without labor (n = 37 each). Placenta growth factor levels were determined with an antigen-capture enzyme-linked immunosorbent assay. RESULTS: Maternal placenta growth factor levels during normal pregnancy increased from the first trimester to the late second trimester; they subsequently declined from 30 weeks' gestation to delivery. Significantly less maternal placenta growth factor (P <.0001) was found in pregnancies complicated by preeclampsia, and labor significantly lowered placenta growth factor levels in both maternal (P =.0189) and cord serum samples (P <.0001). CONCLUSION: Decreased levels of placenta growth factor during preeclampsia could influence endothelial cell and trophoblast function, thereby contributing to the pathogenesis of the disease. PMID- 9855594 TI - Regional variations in contractile responses to prostaglandins and prostanoid receptor messenger ribonucleic acid in pregnant baboon uterus. AB - OBJECTIVES: The aims of this study were to compare (1) the contractile responses of the lower uterine segment and fundus to prostaglandins, (2) expression of genes encoding prostanoid receptors in myo-metrium from different regions of the uterus, and (3) the distribution of expression of genes encoding prostanoid receptors (P receptors) in key intrauterine tissues. STUDY DESIGN: Cesarean hysterectomy was performed in 8 pregnant baboons, not in labor, in the last third of pregnancy. Contractile responses of fresh tissue were quantified in a superfusion system. Polyadenylated ribonucleic acid was extracted from frozen tissue and gene expression was quantified by Northern blot analysis with complementary deoxyribonucleic acid probes. RESULTS: Prostaglandin E2 contracted strips of myometrium from the fundus but had no significant effect on strips from lower uterine segment. Prostaglandin F2 alpha contracted myometrium from both regions equally. Compared with fundus tissue, lower uterine segment tissue had greater expression of EP2 receptor messenger ribonucleic acid, less expression of EP3 receptor messenger ribonucleic acid, but similar levels of EP4 receptor and FP receptor messenger ribonucleic acid. EP2 receptor, EP3 receptor, and EP4 receptor messenger ribonucleic acids were also detected in cervix, decidua, and chorion. EP2 receptor messenger ribonucleic acid was most abundant in the cervix, EP3 receptor messenger ribonucleic acid was most abundant in the myometrium, and EP4 receptor messenger ribonucleic acid was most abundant in the decidua. CONCLUSIONS: The reduced contractile response of lower uterine segment tissue to prostaglandin E2 is paralleled by greater inhibitory EP2 receptor expression and less contractile EP3 receptor expression, a pattern similar to that seen in the cervix. Drugs with selective activity at prostanoid receptor types and subtypes are likely to allow safer and more effective control of the uterus and cervix than native prostaglandins. PMID- 9855595 TI - Fetal fibronectin detection in preterm labor: evaluation of a prototype bedside dipstick technique and cervical assessment. AB - OBJECTIVE: The aims of this study were to evaluate a prototype bedside test for fetal fibronectin detection in women with symptoms of preterm labor, to compare the efficacy of obtaining fetal fibronectin swabs with and without a speculum, and to assess the value of combining the fetal fibronectin test with cervical dilatation for predicting delivery within 10 days. STUDY DESIGN: This investigation prospectively studied a cohort of women with symptoms of preterm labor (n = 121), gestational age between 24 and 33 6/7 weeks, and cervical dilatation /=1 cm predicted delivery within 10 days with sensitivity, specificity, and positive and negative predictive values of 71%, 87%, 46%, and 95%, respectively, with positive and negative likelihood ratios of 5.5 and 0.33, respectively. Combination of fetal fibronectin testing with cervical dilatation >/=1 cm did not significantly improve the fetal fibronectin test characteristics. After exclusion of women with slight vaginal bleeding, cervical dilatation >/=1 cm and fetal fibronectin status remained the only independent variables associated with delivery within 10 days. CONCLUSION: Fetal fibronectin testing according to this prototype may have a limited role in clinical decision analysis. In this study a cervical dilatation >/=1 cm had predictive values equivalent to those of the fetal fibronectin test, and it should be evaluated further in a clinical setting. PMID- 9855596 TI - Vasoactive mediators in pregnancy-induced hypertensive disorders: a longitudinal study. AB - OBJECTIVE: The objective of this study was to evaluate the extent to which endothelin and the eicosanoids prostacyclin and thromboxane A2 are involved in the pathophysiology of gestational hypertension and preeclampsia. STUDY DESIGN: In a longitudinal design, venous blood samples and 24-hour urine specimens were collected from 396 women in each trimester of pregnancy. After delivery of all patients, venous plasma endothelin was assessed in 20 subjects with identified preeclampsia, 48 subjects with gestational hypertension, and 59 normotensive subjects. Urinary excretions of the thromboxane A2 and of the prostacyclin metabolites thromboxane B2 and 6-keto-prostaglandin F1 alpha were assessed in 16 subjects with preeclampsia, 35 subjects with gestational hypertension, and 31 normotensive subjects. RESULTS: Endothelin levels showed a second-trimester drop in all groups. In all 3 gestational trimesters a high correlation was found between the excretion of thromboxane B2 and that of 6-keto-prostaglandin F1 alpha (P <.001). The overall thromboxane B2 and 6-keto-prostaglandin F1 alpha urinary excretions increased throughout pregnancy and the overall thromboxane B2 /6-keto prostaglandin F1 alpha ratio decreased. No significant differences in endothelin, thromboxane B2, and 6-keto-prostaglandin F1 alpha excretion levels or in thromboxane B2 /6-keto-prostaglandin F1 alpha ratios were found between women with preeclampsia, gestational hypertension, and normotension. Only in a small group of patients with severe preeclampsia (n = 2) and severe gestational hypertension (n = 2) were increased second-trimester endothelin values and increased thromboxane B2 /6-keto-prostaglandin F1 alpha ratios found. CONCLUSION: In this longitudinal study we found no evidence for prostacyclin deficiency or increased endothelin levels in preeclampsia. Only women with severe preeclampsia and severe gestational hypertension expressed increased endothelin levels and thromboxane dominance over prostacyclin. PMID- 9855597 TI - Resistance to activated protein C and the leiden mutation: high prevalence in patients with abruptio placentae. AB - OBJECTIVE: The Leiden mutation, a point mutation in the gene encoding coagulation factor V, is associated with a high frequency of thromboembolic phenomena. It has recently been connected with adverse outcomes of pregnancy. We carried out this study to define its connection with abruptio placentae. STUDY DESIGN: Twenty seven women who had abruptio placentae and 29 control subjects matched for age, parity, and ethnic origin were studied. We studied all women for possible hypercoagulation defects. All women demonstrating resistance to activated protein C were studied for the presence of the factor V Leiden mutation. RESULTS: Seventeen of 27 case patients had an activated protein C ratio /=3 spontaneous losses of pregnancy; however, a cause is determined in only 50% of cases. Recent studies have associated the presence of thyroid autoantibodies during the first trimester of pregnancy with spontaneous abortion in the current pregnancy among women without a history of recurrent abortion. The objective of this study was to determine whether circulating thyroid autoantibodies were associated with recurrent pregnancy loss. STUDY DESIGN: Sera from 74 nonpregnant women with a history of recurrent pregnancy loss and from 75 healthy, fertile control subjects of similar gravidity were tested for thyroglobulin and thyroid peroxidase antibodies by means of radioimmunoassay kits. All women had a third-generation thyroid-stimulating hormone assay performed. Samples were obtained >/=6 months after a pregnancy. RESULTS: Twenty two of the women with a history of recurrent pregnancy loss (29.3%) and twenty eight of the control subjects (37%) had positive results for either one or both of the thyroid autoantibodies (P >. 05). Mean thyroid-stimulating hormone levels and the proportion of women with abnormal thyroid-stimulating hormone values did not differ between the 2 groups. CONCLUSION: Women with a history of recurrent pregnancy loss are no more likely than are fertile control subjects to have circulating thyroid autoantibodies. Testing for antithyroid antibodies is not clinically useful in the evaluation of patients with a history of recurrent pregnancy loss. PMID- 9855602 TI - Discordant uterine artery velocity waveforms as a predictor of subsequent miscarriage in early viable pregnancies. AB - OBJECTIVE: Our purpose was to determine whether an abnormal uterine perfusion pattern was associated with subsequent pregnancy loss after fetal cardiac activity was documented. STUDY DESIGN: Pulsatility indexes of both the right and left uterine arteries were obtained by transvaginal color Doppler ultrasonography in 318 consecutive viable pregnancies between 6 and 12 weeks' gestation. The Delta uterine artery pulsatility index value, expressed as the highest uterine artery pulsatility index value minus the lowest value, was calculated for each pregnancy. Women were subsequently classified as having continuing pregnancies or pregnancy loss before 20 weeks' gestation. To predict subsequent pregnancy loss, Doppler findings were adjusted for maternal age, history of previous abortion, presence of subchorionic hematoma, embryonic bradycardia, and gestational age by means of multivariate logistic regression analysis. RESULTS: Twenty-four pregnancies (8%) were spontaneously aborted before 20 weeks' gestation. Both Delta uterine artery pulsatility index (odds ratio 2.9, 95% confidence interval 1.5-5.8) and history of previous abortion (odds ratio 3.1, 95% confidence interval 1.2-8.2) were significantly associated with pregnancy loss in the multivariate logistic regression analysis. The sensitivity and specificity of the multivariate logistic regression model to predict abortion were 75% and 85%, respectively, significantly higher than the diagnostic performances of qualitative and quantitative variables considered individually. CONCLUSION: Discordant uterine artery pulsatility indexes in the first trimester were strongly associated with subsequent pregnancy loss. This suggests that uterine ischemia may be implicated in certain cases of early pregnancy loss after documentation of fetal cardiac activity during the first trimester. PMID- 9855603 TI - Long-term high-altitude hypoxia increases plasma nitrate levels in pregnant ewes and their fetuses. AB - OBJECTIVE: The study was designed to determine whether moderate chronic hypoxia changes plasma nitrate concentrations in nonpregnant and pregnant, near-term ewes and their fetuses. STUDY DESIGN: Pregnant ewes were put into either a normoxic control group or a chronically hypoxic group maintained at high altitude (3820 m, PaO 2 60 mm Hg) from day 30 of gestation. On day 140 blood samples were collected from the maternal jugular vein and from the fetal umbilical artery and vein. Blood samples were also obtained from normoxic nonpregnant adult ewes and nonpregnant adult ewes exposed to high-altitude hypoxia for 110 days. Plasma nitrate concentrations were determined through chemiluminescence assay. RESULTS: Within normoxic groups plasma nitrate concentrations of pregnant, near-term ewes were significantly elevated compared with those of nonpregnant ewes (17.4 +/- 0.3 vs 6.8 +/- 0.4 micromol/L, P <.0001). Fetal umbilical arterial and venous plasma nitrate concentrations did not differ from each other in the normoxic group (44.7 +/- 4.2 and 44.0 +/- 4.3 micromol/L, respectively) but were significantly higher than those of their mothers (P <.0001). Compared with normoxic groups, pregnant ewes with chronic hypoxia showed significantly increased plasma nitrate concentrations (30.9 +/- 1.8 micromol/L, P <.001), as did their fetuses (arterial 146.1 +/- 14.7 micromol/L, venous 154.5 +/- 14.6 micromol/L, P <.0001), but nonpregnant ewes with chronic hypoxia did not (7.5 +/- 0.3 micromol/L, P >.05). CONCLUSIONS: We conclude that (1) production of endogenous nitric oxide is increased by pregnancy in sheep and is higher in the fetus than in the mother, and (2) moderate chronic hypoxia selectively augments nitric oxide production in the pregnant ewe's circulation and, even more profoundly, in the fetal lamb's circulation. PMID- 9855604 TI - Plasma ferritin, premature rupture of membranes, and pregnancy outcome. AB - OBJECTIVE: The objective of this study was to determine whether plasma ferritin levels predict maternal or neonatal outcomes in women with preterm rupture of membranes at <32 weeks' gestation. METHODS: Plasma from 223 women with premature rupture of membranes at <32 weeks' gestation who had participated in a randomized antibiotic trial were analyzed for ferritin at random assignment and at delivery, and the results were compared with the development of clinical chorioamnionitis, latency until delivery, neonatal sepsis, and a composite adverse neonatal outcome variable. RESULTS: The mean plasma ferritin level rose from 19.2 +/- 29.1 microgram/L on admission to 38.3 +/- 54.3 microgram/L at delivery, with a mean latency of 9.3 +/- 14.6 days. Plasma ferritin levels were significantly higher at both times in mothers whose infants acquired sepsis than in those whose infants did not, especially at delivery (68.5 +/- 96.3 microgram/L vs 32.5 +/- 40.5 microgram/L, P =.01), and neonatal sepsis was 2 to 3 times more common among women with plasma ferritin levels above the median than among those with levels below the median. CONCLUSIONS: Among women with premature rupture of membranes at <32 weeks' gestation, plasma ferritin levels were significantly associated with neonatal sepsis. These data suggest that higher plasma ferritin levels may serve as a marker of infection among women with premature rupture of membranes; however, the clinical utility of plasma ferritin levels in predicting neonatal outcome appears limited. PMID- 9855605 TI - Plasma homocysteine concentration is increased in preeclampsia and is associated with evidence of endothelial activation. AB - OBJECTIVE: We tested the hypothesis that the independent risk factor for atherosclerosis of increased plasma homocysteine concentration is associated with the pregnancy syndrome of preeclampsia. We further hypothesized that increased plasma homocysteine concentration during pregnancy may advance endothelial dysfunction in preeclampsia by promoting oxidative stress. STUDY DESIGN: Antepartum blood samples were collected >/=6 hours after the last meal from 33 women with normal, uncomplicated pregnancies and from 21 women with preeclampsia. These plasma samples were analyzed for concentrations of total homocysteine; folate; triglycerides; creatinine; a marker of endothelial activation, cellular fibronectin; and a marker of oxidative stress, malondialdehyde. RESULTS: The mean value of total plasma homocysteine in preeclampsia was significantly higher than that observed in normal pregnancy (P <. 04). Similarly, plasma malondialdehyde (P <.001), triglyceride (P <. 001), and cellular fibronectin (P <.006) concentrations were also greater in women with preeclampsia than in control subjects. However, no differences were observed between women with preeclampsia and control subjects in folate (P =.97) or creatinine (P =.28) concentrations. Homocysteine concentration did not correlate with plasma creatinine (P =.61), malondialdehyde (P =.32), or triglyceride (P =.89) concentrations. However, cellular fibronectin concentration correlated positively with homocysteine concentration in both women with preeclampsia and control subjects (r = 0.87, P <. 0001, and r = 0.50, P <.004, respectively), and folate concentrations were weakly but negatively correlated with homocysteine values (P =.03, r = 0.32). CONCLUSIONS: Total plasma homocysteine concentration is increased in preeclampsia and is significantly correlated with cellular fibronectin concentration, suggesting that homocysteine plays a role in promoting endothelial dysfunction in preeclampsia. Furthermore, despite the use of pregnancy multivitamins and no indications of overt folate deficiency in this subject population, homocysteine concentration weakly and negatively correlates with plasma folate concentration. PMID- 9855606 TI - Abnormal ductus venosus blood flow in trisomy 21 fetuses during early pregnancy. AB - OBJECTIVE: Our purpose was to assess the ductus venosus blood flow in trisomy 21 fetuses in the first half of pregnancy and the relation of this blood flow to nuchal thickness. STUDY DESIGN: The ductus venosus blood flow has been studied in 534 consecutive fetuses between 10 and 18 weeks of gestation who were undergoing prenatal invasive diagnostic procedures. Before the sampling, the velocity during atrial contraction, the pulsatility index for veins, and the ratio of systole-end systole to diastole were established in the Doppler waveforms and eventually correlated with the fetal karyotype. RESULTS: Ductus venosus waveforms were successfully obtained in 82% of the pregnancies. Eleven fetuses were found to have trisomy 21; the pulsatility index was abnormally increased (>95th percentile) in 73% (8/11), the ratio of systole-end-systole to diastole was abnormally increased in 45% (5/11), and the velocity during atrial contraction was decreased (<5th percentile) in 27% (3/11). CONCLUSIONS: An abnormally increased ductus venosus pulsatility index was found in a substantial proportion of early trisomy 21 fetuses, and this was apparently independent of nuchal thickening. PMID- 9855607 TI - Expression of cyclooxygenases in ductus arteriosus of fetal and newborn pigs. AB - OBJECTIVE: We studied the ontogeny of the expression of cyclooxygenase-1 and cyclooxygenase-2 in the ductus arteriosus and evaluated their functional significance. STUDY DESIGN: The expression of cyclooxygenase-1 and cyclooxygenase 2 was studied in ductus arteriosus of fetal (at approximately 75% gestation) and term newborn pig. Effects of selective inhibitors of cyclooxygenase-1 and cyclooxygenase-2 on ductal patency were evaluated by Doppler ultrasonography. RESULTS: Ductus arteriosus of the fetus expressed virtually only cyclooxygenase-1 immunoreactive protein and activity. In contrast, the ductus of the term newborn pig (<45 minutes old) contained proteins of both cyclooxygenase-1 and cyclooxygenase-2, but the latter contributed to >90% of prostaglandin E2 formation. The selective cyclooxygenase-2 inhibitor DuP697 reduced prostaglandin E2 levels in the ductus arteriosus, albeit not in plasma, but did not affect ductal patency in the newborn pig (<1(1/2) hours old); in contrast, the cyclooxygenase-1 inhibitor valeryl salicylate, like indomethacin, markedly reduced levels of prostaglandin E2 in the plasma and ductus arteriosus and caused significant constriction of the ductus arteriosus. CONCLUSION: The ductus arteriosus of the term newborn pig, in contrast to that of the fetus, expresses cyclooxygenase-2, but circulating prostaglandins, arising mostly from cyclooxygenase-1, seem to exert the major control on ductal patency in vivo. Our data suggest that cyclooxygenase-2 inhibitors might be better alternatives for the fetus than nonselective cyclooxygenase blockers if indicated for maternal conditions such as inflammation or for tocolysis. PMID- 9855608 TI - New Down syndrome screening algorithm: ultrasonographic biometry and multiple serum markers combined with maternal age. AB - OBJECTIVE: We compared the Down syndrome screening efficiency of a new algorithm that combines humerus length measurement and serum analytes versus that of the traditional triple-analyte serum screen. STUDY DESIGN: Humerus length measurements were obtained prospectively in 1743 midtrimester (14 to 24 weeks) singleton fetuses before genetic amniocentesis. All patients had triple-marker serum screening before amniocentesis. Data on humerus length were expressed as multiples of the median, and were normalized by log transformation. Backward multiple stepwise logistic regression analysis was performed to determine which combination of biometry and serum markers best predicted fetal Down syndrome. The screening efficiency of the traditional triple-analyte algorithm was compared with that of a new multivariate gaussian algorithm that combined biometry and serum markers. RESULTS: There were 31 (1.8%) fetuses with Down syndrome in the study population. In the regression analysis humerus length, human chorionic gonadotropin, alpha-fetoprotein, and maternal age were significant predictors of Down syndrome, but unconjugated estriol was not. The combined algorithm (humerus length, human chorionic gonadotropin, and alpha-fetoprotein and age) was superior to the traditional triple screen for Down syndrome detection. The sensitivities at fixed false-positive rates were consistently higher in the combination than in the triple-screen protocol. For example, at a 10% false-positive rate the sensitivities were 65.0% and 52.3%, respectively. Similarly, at a 15% false positive rate the sensitivities were 73.5% and 55.0%, respectively. CONCLUSION: A new screening algorithm combining humerus length and serum analytes was superior to the traditional triple screen. Although we used a high-risk population in this study, it is expected that the observed superiority of the combination screen would persist in a population of younger women. The development of a combined biometric and serum analyte screening algorithm for estimating individual odds could represent an advance in prenatal Down syndrome screening. PMID- 9855609 TI - Very low birth weight outcomes of the National Institute of Child Health and Human Development Neonatal Research Network, January 1993 through December 1994. AB - OBJECTIVES: Our purpose was to determine the mortality and morbidity rates for infants weighing 501 to 1500 g according to gestational age, birth weight, and gender. STUDY DESIGN: Perinatal data were collected prospectively on an inborn cohort from January 1993 through December 1994 by 12 participating centers of the National Institute of Child Health and Human Development Neonatal Research Network and were compared with the corresponding data from previous reports. Sociodemographic factors, perinatal events, and the neonatal course to 120 days of life, discharge, or death were evaluated. RESULTS: Eighty-three percent of infants survived until discharge to home or to a long- term care facility (compared with 74% in 1988). Survival to discharge was 49% for infants weighing 501 to 750 g at birth, 85% for those 751 to 1000 g, 93% for those 1001 to 1250 g, and 96% for those 1251 to 1500 g. The majority of deaths occurred within the first 3 days of life. Mortality rates were greater for male than for female infants. Respiratory distress syndrome was the most frequent pulmonary disease (52%). Chronic lung disease (defined as an oxygen requirement at 36 weeks after conception) developed in 19%. Thirty-two percent of infants had evidence of intracranial hemorrhage. Periventricular leukomalacia was noted in 6% of infants who had ultrasonography after 2 weeks. The average duration of hospitalization for survivors was 68 days (122 days for surviving infants weighing 501 to 750 g, compared with an average of 43 days for surviving infants 1251 to 1500 g). Among infants who died, the average length of stay was 19 days. CONCLUSIONS: The mortality rate for infants weighing between 501 and 1500 g at birth continues to decline. This increase in survival is not accompanied by an increase in medical morbidity. There are interactions between birth weight, gestational age, sex, and survival rates. PMID- 9855610 TI - Meconium has no lecithin or sphingomyelin but affects the lecithin/sphingomyelin ratio. AB - OBJECTIVE: Although a few studies have evaluated the effect of meconium on the lecithin/sphingomyelin ratio for testing of fetal lung maturity, to date these studies have assessed only the lecithin-sphingomyelin ratio of amniotic fluid contaminated with meconium. The purpose of this study was (1) to determine whether meconium by itself has a lecithin/sphingomyelin ratio and, if so, (2) to determine whether the lecithin/sphingomyelin ratio is constant. STUDY DESIGN: A lecithin/sphingomyelin ratio was obtained by standard thin-layer chromatography on the first meconium stool of 20 neonates between 31 weeks and term. A quantitative assay was then performed on a sample from each gestational age (7 samples ranging from 31 weeks to term) to confirm the presence of lecithin and sphingomyelin. RESULTS: The 20 samples had atypical thin-layer chromatographic migratory patterns in the zones for lecithin and sphingomyelin. The presumed lecithin/sphingomyelin ratios ranged from 1.1 to 3.6, with no correlation with gestational age. However, the quantitative assay did not detect the presence of lecithin or sphingomyelin in any of the analyzed samples. CONCLUSIONS: Meconium does not appear to contain lecithin or sphingomyelin but has an unidentified moiety whose migratory pattern, as shown by qualitative standard thin-layer chromatography, is similar to that of lecithin with sphingomyelin. Therefore the presence of meconium in amniotic fluid may falsely raise or lower the lecithin/sphingomyelin ratio and confuse fetal lung maturity interpretations. PMID- 9855611 TI - Outcome of cardiovascular surgery and pregnancy: a systematic review of the period 1984-1996. AB - The outcomes of cardiovascular operations during pregnancy, at delivery, and post partum were reviewed from published material in the period 1984-1996. Surgery during pregnancy resulted in fetal-neonatal morbidity and mortality of 9% and 30%, respectively, and in maternal morbidity and mortality of 24% and 6%, respectively. Duration of pregnancy at surgery and duration and temperature of cardiopulmonary bypass did not influence fetal-neonatal outcome. Maternal complications and mortality of surgery immediately after delivery were 29% and 12%, respectively, and for surgery performed with a postpartum interval the respective rates were 38% and 14%. Hospitalization after week 27 of gestation and extreme emergency contributed significantly to poor maternal outcome. Maternal deaths were reported in 9% of valvular procedures and in 22% of aortic or arterial dissection repairs and pulmonary embolectomies. Fetal-neonatal risks of maternal surgery during pregnancy are high and unpredictable. Maternal risks of cardiovascular procedures during pregnancy are moderate, significantly increase if an operation is performed at or after delivery, and, overall, should be considered as higher than those in nonpregnant cardiovascular surgical patients. PMID- 9855612 TI - An abortus with double trisomy (48,XY,+4,+7) PMID- 9855613 TI - Polycystic ovary syndrome: metabolic challenges and new treatment options. PMID- 9855614 TI - Polycystic ovary syndrome: symptomatology, pathophysiology, and epidemiology. AB - Women with polycystic ovary syndrome seek health care for 3 major reasons: infertility, menstrual irregularity, and androgen excess. The infertility is associated with anovulation. The menstrual irregularity is typically chronic, beginning with menarche. Although amenorrhea may sometimes occur, the more common presentation is irregular bleeding characteristic of anovulation. Androgen excess may be manifested by varying degrees of hirsutism. Patients may also report acne. The rapid development of virilizing signs, such as deepening of the voice, increased muscle mass, and temporal balding, should prompt a search for a tumor and lead one away from a diagnosis of polycystic ovary syndrome. Typically treatment is directed at alleviating the symptoms: ovulation induction for infertility, oral contraceptives or a progestin for menstrual irregularity, and oral contraceptives or spironolactone for hirsutism. On the basis of recent epidemiologic data suggestive of increased cardiovascular risk among women with polycystic ovary syndrome, such treatment might be complemented by a long-term approach that addresses the underlying pathophysiology of insulin resistance. PMID- 9855615 TI - Understanding the underlying metabolic abnormalities of polycystic ovary syndrome and their implications. AB - Women with polycystic ovary syndrome have an increased rate of obesity, with a propensity toward abdominal deposition of body fat. Independent of obesity, at least half of affected women have insulin resistance. To understand the mechanisms of insulin resistance in polycystic ovary syndrome, it is necessary to understand normal insulin signaling. Women with polycystic ovary syndrome have normal binding of insulin to its receptor but have decreased activation of events downstream of the receptor. This insulin resistance occurs mostly in the peripheral tissues (muscle and fat cells), and results in increased pancreatic insulin secretion to maintain normal glucose levels. Obese women with polycystic ovary syndrome have a relative defect in insulin secretion. At least 20% of obese patients with polycystic ovary syndrome have glucose intolerance or diabetes, versus about 5% of the healthy age-matched population. Obesity and insulin resistance probably increase the risk of cardiovascular disease in women with polycystic ovary syndrome. The metabolic features of polycystic ovary syndrome are important health risk factors and need to be considered seriously, even if the patient seeks treatment for other concerns. PMID- 9855616 TI - Polycystic ovary syndrome: current and future treatment paradigms. AB - Polycystic ovary syndrome is characterized by excess levels of circulating androgens and by chronic anovulation. Although the fundamental pathophysiologic defect has not been determined, women with polycystic ovary syndrome are known to be uniquely insulin resistant. Obesity in polycystic ovary syndrome aggravates the underlying predisposition towards insulin resistance. Diagnostic criteria that focus on menstrual irregularity are more likely to discriminate insulin resistant women than are such criteria as abnormal gonadotropin secretion or ovarian morphologic characteristics. About 40% of patients with polycystic ovary syndrome demonstrate glucose intolerance (either impaired glucose tolerance or type 2 diabetes) in response to an oral glucose challenge. The lack of a clear causal mechanism in the syndrome has led to a multitude of symptom-oriented treatments, with few therapies improving all aspects of the endocrine abnormalities associated with polycystic ovary syndrome. Many of these therapies such as ovulation induction with medical agents-hold increased risks for women with polycystic ovary syndrome, including ovarian hyperstimulation syndrome and multiple gestation. Empirical studies of interventions that improve insulin sensitivity in polycystic ovary syndrome (either weight loss and diet programs or pharmaceutical agents) have been shown to improve the endocrine abnormalities in the syndrome. Initial results with antidiabetic agents (specifically insulin sensitizing agents) are promising but need to be confirmed with larger, randomized studies. PMID- 9855617 TI - The obstetrician-gynecologist's role in the practical management of polycystic ovary syndrome. AB - Women with polycystic ovary syndrome come to the gynecologist with a variety of symptoms, including menstrual irregularities, hirsutism, acne, weight gain, obesity, and infertility. An accurate diagnosis requires both confirmation of signs and symptoms of polycystic ovary syndrome and exclusion of other disorders. Once the diagnosis of polycystic ovary syndrome has been established, the presence of concomitant conditions, such as hypertension, dyslipidemia, and diabetes, must be assessed. Because the cause of polycystic ovary syndrome is not clear, treatment options have focused on symptom management. Such treatment options include oral contraceptives, gonadotropin-releasing hormone analogs with "add-back" hormone regimens, antiandrogens, ovulation-inducing agents, electrolysis, nutritional and weight loss counseling, exercise, laparoscopic ovarian drilling, and glucocorticoids. Pathogenic considerations, risk factor assessments, and treatment objectives combine to determine the choice of therapies. It is not clear whether insulin resistance is clinically important or causal in polycystic ovary syndrome symptom complex in all affected women. Polycystic ovary syndrome may be the final common expression of a variety of metabolic or neuroendocrine perturbations. If insulin resistance is a universal feature, it would make sense to treat with an insulin-sensitizing agent in the expectation that symptoms would resolve or improve. If insulin resistance is not the main etiologic factor, however, then insulin-sensitizing agents would be useful as adjunctive agents only for women with clinically important insulin resistance (eg, patients with polycystic ovary syndrome in whom insulin resistance causes hyperglycemia). In such cases an insulin-sensitizing agent could be instituted along with a program of weight loss and exercise. PMID- 9855619 TI - Agonist-induced phosphorylation of the angiotensin AT1a receptor is localized to a serine/threonine-rich region of its cytoplasmic tail. AB - The agonist-induced phosphorylation sites of the rat AT1a angiotensin receptor were analyzed using epitope-tagged mutant receptors expressed in Cos-7 cells. Angiotensin II-stimulated receptor phosphorylation was unaffected by truncation of the cytoplasmic tail of the receptor at Ser342 (Delta342) but was abolished by truncation at Ser325 (Delta325). Truncation at Ser335 (Delta335), or double-point mutations of Ser335 and Thr336 to alanine (ST-AA), reduced receptor phosphorylation by approximately 50%, indicating that in addition to Ser335 and/or Thr336, amino acids within the Ser326-Thr332 segment are also phosphorylated. Agonist-induced phosphorylation of the ST-AA and Delta335 receptors was partially inhibited by staurosporine, suggesting that the single protein kinase C consensus site in the Ser326-Thr332 segment (Ser331) is phosphorylated. The impairment of receptor phosphorylation was broadly correlated with the attenuation of agonist-induced internalization rates (Delta325 < Delta335 < ST-AA < Delta342 < wild-type) and with the increasing rank order of magnitude of inositol phosphate production normalized to an equal number of receptors (Delta325 > Delta335 > ST-AA = Delta342 > wild-type). These results demonstrate that agonist-induced phosphorylation of the AT1a receptor is confined to an 11-amino-acid serine/threonine-rich segment of its carboxyl-terminal cytoplasmic tail and implicate this region in the mechanisms of receptor internalization and desensitization. PMID- 9855620 TI - A single amino acid, glu146, governs the substrate specificity of a human dopamine sulfotransferase, SULT1A3. AB - Sulfation, catalyzed by members of the sulfotransferase (SULT) superfamily, exerts considerable influence over the biological activity of numerous endogenous and xenobiotic chemicals. In humans, catecholamines such as dopamine are extensively sulfated, and a SULT isoform (SULT1A3 or the monoamine-sulfating form of phenolsulfotransferase) has evolved with considerable selectivity for dopamine and other biogenic amines. To investigate the molecular basis for this selectivity, we identified a region of SULT1A3, which, we hypothesized, contributes to its preference for biogenic amines, and mutated two amino acids within this domain to the corresponding residues in a closely related but functionally distinct phenol sulfotransferase, SULT1A1 (H143Y and E146A). The change of a single amino acid, E146A, was sufficient to transform the catalytic properties and substrate preference of SULT1A3, such that they closely resembled those of SULT1A1. These experiments confirm the functional role of Glu146 in the selectivity of SULT1A3 for biogenic amines and suggest that this region is a key determinant of sulfotransferase substrate specificity. PMID- 9855621 TI - Chronic morphine augments adenylyl cyclase phosphorylation: relevance to altered signaling during tolerance/dependence. AB - Despite the demonstration that chronic morphine increases phosphorylation of multiple substrate proteins, their identity has, for the most part, remained elusive. Thus far, chronic morphine has not been shown to increase the phosphorylation of any identified effector protein. This is the first demonstration that persistent activation of opioid receptors has profound effects on phosphorylation of adenylyl cyclase (AC). A dramatic increase in phosphorylation of AC (type II family) was observed in ileum longitudinal muscle myenteric plexus preparations obtained from chronic morphine-treated guinea pigs. Analogous results were obtained when AC was immunoprecipitated using two differentially directed AC antibodies. The magnitude of the augmented AC phosphorylation was substantially attenuated by chelerythrine, a protein kinase C selective inhibitor. These results suggest the potential relevance of increased phosphorylation (protein kinase C-mediated) of AC to opioid tolerant/dependent mechanisms. Because phosphorylation of AC isoforms (type II family) can significantly increase their stimulatory responsiveness to Gsalpha and Gbetagamma, this mechanism could underlie, in part, the predominance of opioid AC stimulatory signaling observed in opioid tolerant/dependent tissue. Moreover, in light of the fact that many G protein-coupled receptors signal through common effector proteins, this effect provides a mechanism for divergent consequences of chronic morphine treatment and could explain the well documented complexity of changes that accompany the opioid tolerant/dependent state. PMID- 9855622 TI - An evolutionarily conserved cysteine protease, human bleomycin hydrolase, binds to the human homologue of ubiquitin-conjugating enzyme 9. AB - Bleomycin hydrolase (BH) is a highly conserved cysteine proteinase that deamidates and inactivates the anticancer drug bleomycin. Yeast BH self-assembles to form a homohexameric structure, which resembles a 20 S proteasome and may interact with other proteins. Therefore, we searched for potential human BH (hBH) partners using the yeast two-hybrid system with a HeLa cDNA library and identified the full-length human homologue of yeast ubiquitin-conjugating enzyme 9 (UBC9). Cotransformation assays using hBH deletion mutants revealed that the carboxyl terminus of hBH (amino acids 356-455), which contains two of the three essential catalytic amino acids, was not critical for protein binding in the yeast two-hybrid environment. In vitro translated human UBC9 was precipitated by glutathione S-transferase-hBH fusion protein but not by glutathione S transferase. Efficient in vitro binding occurred in the absence of the first 24 amino acids of UBC9 and the catalytic Cys93 of UBC9. We confirmed that hBH and UBC9 interacted in vivo by affinity copurification of proteins overexpressed in mammalian cells. Using immunocytochemical analysis, hBH was colocalized with UBC9. Coexpression of hBH and UBC9 in mammalian cells did not markedly alter the bleomycin-hydrolyzing activity of hBH or apparent small ubiquitin-related modifier 1 addition. This is the first reported heteromeric interaction with hBH, and it suggests a role for hBH in intracellular protein processing and degradation. PMID- 9855623 TI - YC-1 potentiates nitric oxide- and carbon monoxide-induced cyclic GMP effects in human platelets. AB - Nitric oxide (NO), the physiological activator of soluble guanylyl cyclase (sGC), induces inhibitory effects on platelet activation via elevation of cGMP levels and stimulation of the cGMP-dependent protein kinase. YC-1, a benzylindazole derivative, was shown to activate sGC in intact platelets, resulting in inhibition of platelet aggregation. In a previous study, we demonstrated that YC 1 not only stimulates purified sGC but also potentiates the stimulatory action of submaximally effective NO and carbon monoxide (CO) concentrations. Here, we investigated the potentiating effect of YC-1 in intact platelets. YC-1 together with NO or CO led to complete inhibition of platelet aggregation at concentrations that were ineffective by themselves. Maximally effective 2, 2 diethyl-1-nitroso-oxyhydrazine (3 microM) and YC-1 (100 microM) concentrations each elevated the cGMP levels in intact platelets approximately 13-fold, and administration of the two drugs together resulted in enormous potentiation of cGMP formation, which greatly exceeded the effect on the purified enzyme and yielded a >1300-fold increase in cGMP levels. Similar results were obtained using CO instead of NO. Furthermore, YC-1 not only stimulated sGC but also inhibited cGMP-hydrolyzing phosphodiesterases in platelets. The enormous elevation of cGMP levels led to enhanced phosphorylation of the cGMP-dependent protein kinase substrate vasodilator-stimulated phosphoprotein. Thus, by the combination of two effects (i.e., potentiation of NO-induced sGC stimulation and phosphodiesterase inhibition), YC-1-like substances are potent activators of the sGC/cGMP pathways and are therefore interesting candidates to act as modulators of cGMP-mediated effects, especially within the cardiovascular system. PMID- 9855624 TI - A hydrophobic cluster between transmembrane helices 5 and 6 constrains the thyrotropin-releasing hormone receptor in an inactive conformation. AB - We have studied the role of a highly conserved tryptophan and other aromatic residues of the thyrotropin-releasing hormone (TRH) receptor (TRH-R) that are predicted by computer modeling to form a hydrophobic cluster between transmembrane helix (TM)5 and TM6. The affinity of a mutant TRH-R, in which Trp279 was substituted by alanine (W279A TRH-R), for most tested agonists was higher than that of wild-type (WT) TRH-R, whereas its affinity for inverse agonists was diminished, suggesting that W279A TRH-R is constitutively active. We found that W279A TRH-R exhibited 3.9-fold more signaling activity than WT TRH-R in the absence of agonist. This increased basal activity was inhibited by the inverse agonist midazolam, confirming that the mutant receptor is constitutively active. Computer-simulated models of the unoccupied WT TRH-R, the TRH-occupied WT TRH-R, and various TRH-R mutants predict that a hydrophobic cluster of residues, including Trp279 (TM6), Tyr282, and Phe199 (TM5), constrains the receptor in an inactive conformation. In support of this model, we found that substitution of Phe199 by alanine or of Tyr282 by alanine or phenylalanine, but not of Tyr200 (by alanine or phenylalanine), resulted in a constitutively active receptor. We propose that a hydrophobic cluster including residues in TM5 and TM6 constrains the TRH-R in an inactive conformation via interhelical interactions. Disruption of these constraints by TRH binding or by mutation leads to changes in the relative positions of TM5 and TM6 and to the formation of an active form of TRH R. PMID- 9855625 TI - Naturally occurring analogs of lysophosphatidic acid elicit different cellular responses through selective activation of multiple receptor subtypes. AB - Lysophosphatidic acid (LPA), plasmalogen-glycerophosphate (alkenyl-GP) and, cyclic-phosphatidic acid (cyclic-PA) are naturally occurring phospholipid growth factors (PLGFs). PLGFs elicit diverse biological effects via the activation of G protein-coupled receptors in a variety of cell types. In NIH3T3 fibroblasts, LPA and alkenyl-GP both induced proliferation, whereas cyclic-PA was antiproliferative. LPA and alkenyl-GP decreased cAMP in a pertussis toxin sensitive manner, whereas cyclic-PA caused cAMP to increase. LPA and alkenyl-GP both stimulated the activity of the mitogen-actived protein kinases extracellular signal regulated kinases 1 and 2 and c-Jun NH2-terminal kinase, whereas cyclic-PA did not. All three PLGFs induced the formation of stress fibers in NIH3T3 fibroblasts. To determine whether these lipids activated the same or different receptors, heterologous desensitization patterns were established among the three PLGFs by monitoring changes in intracellular Ca2+ in NIH3T3 fibroblasts. LPA cross-desensitized both the alkenyl-GP and cyclic-PA responses. Alkenyl-GP cross desensitized the cyclic-PA response, but only partially desensitized the LPA response. Cyclic-PA only partially desensitized both the alkenyl-GP and LPA responses. We propose that pharmacologically distinct subsets of PLGF receptors exist that distinguish between cyclic-PA and alkenyl-GP, but are all activated by LPA. We provide evidence that the PSP24 receptor is selective for LPA and not activated by the other two PLGFs. RT-PCR and Northern blot analysis indicate the co-expression of mRNAs encoding the EDG-2, EDG-4, and PSP24 receptors in a variety of cell lines and tissues. However, the lack of mRNA expression for these three receptors in the LPA-responsive Rat-1 and Sp2-O-Ag14 cells suggests that a number of PLGF receptor subtypes remain unidentified. PMID- 9855626 TI - Co-expression of P2X1 and P2X5 receptor subunits reveals a novel ATP-gated ion channel. AB - P2X receptors are a family of ion channels gated by extracellular ATP. Each member of the family can form functional homomeric channels, but only P2X2 and P2X3 have been shown to combine to form a unique heteromeric channel. Data from in situ hybridization studies suggest that P2X1 and P2X5 may also co-assemble. In this study, we tested this hypothesis by expressing recombinant P2X1 and P2X5 receptor subunits either individually or together in human embryonic kidney 293 cells. In cells expressing the homomeric P2X1 receptor, 30 microM alpha,beta methylene ATP (alpha,beta-me-ATP) evoked robust currents that completely desensitized in less than 1 sec, whereas alpha,beta-me-ATP failed to evoke current in cells expressing the homomeric P2X5 receptor. By contrast, alpha, beta me-ATP evoked biphasic currents with a pronounced nondesensitizing plateau phase in cells that co-expressed both subunits. Further, the EC50 for alpha,beta-me-ATP was greater in cells expressing both P2X1 and P2X5 than in cells expressing P2X1 alone (5 and 1.6 microM, respectively). Heteromeric assembly was confirmed using a co-immunoprecipitation assay of epitope-tagged P2X1 and P2X5 subunits. In summary, this study provides biochemical and functional evidence of a novel channel formed by P2X subunit heteropolymerization. This finding suggests that heteromeric subunit assembly constitutes an important mechanism for generating functional diversity of ATP-mediated responses. PMID- 9855627 TI - Diospyrin, a bisnaphthoquinone: a novel inhibitor of type I DNA topoisomerase of Leishmania donovani. AB - Diospyrin is a plant product that has significant inhibitory effect on the growth of Leishmania donovani promastigotes. This compound inhibits the catalytic activity of DNA topoisomerase I of the parasite. Like camptothecin, it induces topoisomerase I mediated DNA cleavage in vitro. Treatment of DNA with diospyrin before addition of topoisomerase I has no effect. Preincubation of topoisomerase I with diospyrin before the addition of DNA in the relaxation reaction increases this inhibition. Our results suggest that this bis-naphthoquinone compound exerts its inhibitory effect by binding with the enzyme and stabilizing the topoisomerase I-DNA "cleavable complex." Diospyrin is a specific inhibitor of the parasitic topoisomerase I. It does not inhibit type II topoisomerase of L. donovani and requires much higher concentrations to inhibit type I topoisomerase of calf thymus. The potent inhibitory effect of diospyrin on type I DNA topoisomerase from L. donovani can be exploited for rational drug design in human leishmaniasis. PMID- 9855628 TI - Bone marrow stromal cells constitutively express high levels of cytochrome P4501B1 that metabolize 7,12-dimethylbenz[a]anthracene. AB - The polycyclic aromatic hydrocarbon 7,12-dimethylbenz[a]anthracene (DMBA) is a potent carcinogen that produces immunotoxic effects in bone marrow. Here, we show that bone marrow stromal cells metabolize DMBA to such products as 3,4 dihydrodiol, the precursor to the most mutagenic DMBA metabolite. The BMS2 bone marrow stromal cell line constitutively expressed higher levels of CYP1B1 protein and mRNA than C3H10T1/2 mouse embryo fibroblasts. BMS2 cells also produced a DMBA metabolite profile that was consistent with CYP1B1 activity. Treatment with the potent aryl hydrocarbon receptor (AhR) ligand 2,3, 7,8-tetrachlorodibenzo-p dioxin (TCDD) induced a approximately 2-fold increase in CYP1B1 mRNA, protein, and activity in BMS2 cells. Two forms of the AhR (97 and 104 kDa) and the AhR nuclear translocator were detected in BMS2 cells. The AhR translocated to the nucleus after treatment with TCDD or DMBA but was approximately 5 times slower with DMBA. Primary bone marrow stromal (BMS) cell cultures established from AhR-/ mice showed similar basal CYP1B1 expression and activity as cell cultures established from heterozygous littermates or C57BL/6 mice. However, primary BMS cells from AhR-/- mice did not exhibit increased CYP1B1 protein expression after incubation with TCDD. BMS cells therefore constitutively express functional CYP1B1 that is not dependent on the AhR. This contrasts with embryo fibroblasts from the same mouse strain, in which basal CYP1B1 expression is AhR dependent. We therefore conclude that bone marrow toxicity may be mediated by CYP1B1-dependent DMBA metabolism, which is regulated by factors other than the AhR. PMID- 9855629 TI - Platelet-derived growth factor-BB and thrombin activate phosphoinositide 3-kinase and protein kinase B: role in mediating airway smooth muscle proliferation. AB - Proliferation of airway smooth muscle results from persistent inflammatory cytokine and growth factor stimulation and is a critical component of airway luminal narrowing in chronic asthma. Using primary cultures of bovine tracheal smooth muscle (BTSM) cells to examine the signaling basis of cell proliferation, platelet-derived growth factor (PDGF)-BB and thrombin (which act through distinct receptor types) were found to induce DNA synthesis in BTSM cells. Mitogen-induced DNA synthesis could be completely inhibited by LY294002, a selective phosphoinositide 3-kinase (PtdIns 3-kinase) inhibitor. Exposure of BTSM cells to PDGF-BB or thrombin resulted in rapid activation of PtdIns 3-kinase and accumulation of phosphoinositide-3,4,5-trisphosphate. Protein kinase B, a novel signaling protein kinase, was identified in BTSM cells and was activated by PDGF BB and thrombin in a PtdIns 3-kinase-dependent manner; this may underlie mitogen stimulated activation of p70(s6k). PD98059, a mitogen-activated protein kinase kinase 1 inhibitor, also partially inhibited PDGF-BB- and thrombin-stimulated DNA synthesis, indicating a modulatory role for mitogen-activated protein kinase in proliferation. GF109203X, Ro 31-8220, calphostin C, and chelerythrine (selective protein kinase C inhibitors) had no effect on PDGF-BB- or thrombin-stimulated DNA synthesis, suggesting that, despite abolishment of mitogen-stimulated protein kinase C activity, cell proliferation stimulated by PDGF-BB and thrombin is protein kinase C-independent. These data demonstrate that the PtdIns 3 kinase/protein kinase B pathway represents a key signaling route in airway smooth muscle proliferation, with the mitogen-activated protein kinase kinase 1/mitogen activated protein kinase cascade providing a complementary signal required for the full mitogenic response. PMID- 9855630 TI - Identification of a glucocorticoid response element in the rat beta2-adrenergic receptor gene. AB - Regulation of beta2-adrenergic receptor (beta2AR) levels by glucocorticoids is a physiologically important mechanism for altering beta2AR responsiveness. Glucocorticoids increase beta2AR density by increasing the rate of beta2AR gene transcription, but the cis-elements involved have not been well characterized. We now show that one of six potential glucocorticoid response elements (GREs) in the 5'-flanking region of the rat beta2AR gene is necessary for glucocorticoid dependent stimulation of receptor gene expression. Using a nested set of deletion fragments of the rat beta2AR gene 5'-flanking region fused to a luciferase reporter gene, glucocorticoid-dependent induction of reporter gene expression in HepG2 cells was localized to a region between positions -643 and -152, relative to the transcription initiation site. In electrophoretic mobility shift assays, a double-stranded oligonucleotide incorporating a near-consensus GRE from this region (positions -379 to -365) formed complexes with the human recombinant glucocorticoid receptor, as well as with nuclear protein from dexamethasone treated HepG2 cells. Mutation of a single base within this GRE sequence greatly diminished interaction of the mutated oligonucleotide with the human recombinant glucocorticoid receptor. The functional activity of the GRE was characterized using a luciferase reporter construct driven by a minimal thymidine kinase promoter. In HepG2 cells transfected with constructs containing the GRE, dexamethasone increased reporter gene expression approximately 3-fold, whereas a dexamethasone effect was not observed with constructs lacking the GRE. Taken together, these findings show that a GRE located at positions -379 to -365 in the 5'-flanking region of the rat beta2AR gene mediates glucocorticoid stimulation of beta2AR gene transcription. PMID- 9855631 TI - Oxidative stress and cytotoxicity induced by ferric-nitrilotriacetate in HepG2 cells that express cytochrome P450 2E1. AB - Iron can potentiate the toxicity of ethanol. Ethanol increases the content of cytochrome P450 2E1 (CYP2E1), which generates reactive oxygen species, and transition metals such as iron are powerful catalysts of hydroxyl radical formation and lipid peroxidation. Experiments were carried out to attempt to link CYP2E1, iron, and oxidative stress as a potential mechanism by which iron increases ethanol toxicity. The addition of ferric-nitrilotriacetate (Fe-NTA) to a HepG2 cell line expressing CYP2E1 decreased cell viability, whereas little effect was observed in control cells not expressing CYP2E1. Toxicity in the CYP2E1-expressing cells was markedly enhanced after the depletion of glutathione. Lipid peroxidation was increased by Fe-NTA, especially in cell extracts and medium from the CYP2E1-expressing cells. Toxicity was completely prevented by vitamin E or by 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, which also decreased the lipid peroxidation. Levels of ATP were lowered by Fe-NTA, and this was associated with a decreased rate of oxygen consumption by permeabilized cells with substrates donating electrons to complexes I, II, and IV of the respiratory chain. This mitochondrial damage was prevented by vitamin E. Toxicity was accompanied by DNA fragmentation, and this fragmentation was prevented by antioxidants. Overexpression of bcl-2 decreased the toxicity and DNA fragmentation produced by the combination of CYP2E1 plus Fe-NTA, as did a peptide inhibitor of caspase 3. These results suggest that elevated generation of reactive oxygen species in HepG2 cells expressing CYP2E1 leads to lipid peroxidation in the presence of iron, and the ensuing prooxidative state damages mitochondria, releasing factors that activate caspase 3, leading to a loss in cell viability and DNA fragmentation. PMID- 9855632 TI - DNA-binding preferences of bisantrene analogues: relevance to the sequence specificity of drug-mediated topoisomerase II poisoning. AB - To elucidate structure-activity relationships for drugs that are able to poison or inhibit topoisomerase II, we investigated the thermodynamics and stereochemistry of the DNA binding of a number of anthracene derivatives bearing one or two 4, 5-dihydro-1H-imidazol-2-yl-hydrazone side chains (characteristic of bisantrene) at different positions of the planar aromatic system. An aza bioisostere, which can be considered a bisantrene-amsacrine hybrid, was also tested. The affinity for nucleic acids in different sequence contexts was evaluated by spectroscopic techniques, using various experimental conditions. DNA melting and DNase I footprinting experiments were also performed. The location and number of the otherwise identical side chains dramatically affected the affinity of the test compounds for the nucleic acid. In addition, the new compounds exhibited different DNA sequence preferences, depending on the locations of the dihydroimidazolyl-hydrazone groups, which indicates a major role for the side-chain position in generating specific contacts with the nucleic acid. Molecular modeling studies of the intercalative binding of the 1- or 9 substituted isomers to DNA fully supported the experimental data, because a substantially more favorable recognition of A-T steps, compared with G-C steps, was found for the 9-substituted derivative, whereas a much closer energy balance was found for the 1-substituted isomer. These results compare well with the alteration of base specificity found for the topoisomerase II-mediated DNA cleavage stimulated by the isomeric drugs. Therefore, DNA-binding specificity appears to represent an important determinant for the recognition of the topoisomerase-DNA cleavable complex by the drug, at least for poisons belonging to the amsacrine-bisantrene family. PMID- 9855633 TI - Protective effects of the antiparkinsonian drugs talipexole and pramipexole against 1-methyl-4-phenylpyridinium-induced apoptotic death in human neuroblastoma SH-SY5Y cells. AB - Treatment of human neuroblastoma SH-SY5Y cells with 1 mM 1-methyl-4 phenylpyridinium (MPP+) for 3 days induced production of reactive oxygen species (ROS), followed by caspase-3 activation, cleavage of poly(ADP-ribose) polymerase (PARP), and apoptotic cell death with DNA fragmentation and characteristic morphological changes (condensed chromatin and fragmented nuclei). Simultaneous treatment with 1 mM talipexole slightly inhibited the MPP+-induced ROS production and apoptotic cell death. In contrast, pretreatment with 1 mM talipexole for 4 days markedly protected the cells against MPP+-induced apoptosis. However, this protective effect might not be mediated by dopamine receptors. The talipexole pretreatment induced an increase in antiapoptotic Bcl-2 protein level but had no effect on levels of proapoptotic Bax, Bak, and Bad. It also inhibited MPP+ induced ROS production, p53 expression, and cleavages of caspase-3 and PARP. Similarly, pramipexole pretreatment increased Bcl-2 and inhibited MPP+-induced apoptosis. Although pretreatment with bromocriptine also had a protective effect against MPP+-induced apoptosis, it had no effect on the protein levels of Bcl-2 family members. On the other hand, N6,2'-O-dibutyryl cAMP or calphostin C induced a decreased Bcl-2 level and enhanced MPP+-induced cell death. These results suggest that talipexole has dual actions: (1) it directly scavenges ROS, affording slight protection against MPP+-induced apoptosis, and (2) it induces Bcl-2 expression, thereby affording more potent protection, if it is administrated before MPP+. Pramipexole has similar effects, whereas bromocriptine seems to exhibit the former but not the latter effect. PMID- 9855634 TI - Regulation of N-methyl-D-aspartate receptor function by constitutively active protein kinase C. AB - The ability of the constitutively active fragment of protein kinase C (PKM) to modulate N-methyl-D-aspartate (NMDA)-activated currents in cultured mouse hippocampal neurons and acutely isolated CA1 hippocampal neurons from postnatal rats was studied using patch-clamp techniques. The responses of two heterodimeric combinations of recombinant NMDA receptors (NR1a/NR2A and NR1a/NR2B) expressed in human embryonic kidney 293 cells were also examined. Intracellular applications of PKM potentiated NMDA-evoked currents in cultured and isolated CA1 hippocampal neurons. This potentiation was observed in the absence or presence of extracellular Ca2+ and was prevented by the coapplication of the inhibitory peptide protein kinase inhibitor(19-36). Furthermore, the PKM-induced potentiation was not a consequence of a reduction in the sensitivity of the currents to voltage-dependent blockade by extracellular Mg2+. We also found different sensitivities of the responses of recombinant NMDA receptors to the intracellular application of PKM. Some potentiation was observed with the NR1a/NR2A subunits, but none was observed with the NR1a/NR2B combination. Applications of PKM to inside-out patches taken from cultured neurons increased the probability of channel opening without changing single-channel current amplitudes or channel open times. Thus, the activation of protein kinase C is associated with potentiation of NMDA receptor function in hippocampal neurons largely through an increase in the probability of channel opening. PMID- 9855635 TI - SR 141716A acts as an inverse agonist to increase neuronal voltage-dependent Ca2+ currents by reversal of tonic CB1 cannabinoid receptor activity. AB - The CB1 cannabinoid receptor antagonist SR 141716A abolished the inhibition of Ca2+ currents by the agonist WIN 55,212-2. However, SR 141716A alone increased Ca2+ currents, with an EC50 of 32 nM, in neurons that had been microinjected with CB1 cRNA. For an antagonist to elicit an effect, some receptors must be tonically active. Evidence for tonically active CB1 receptors was seen as enhanced tonic inhibition of Ca2+ currents. Preincubation with anandamide failed to enhance the effect of SR 141716A, indicating that anandamide did not cause receptor activity. Under Ca2+-free conditions designed to block the Ca2+-dependent formation of anandamide and sn-2-arachidonylglycerol, SR 141716A again increased the Ca2+ current. The Ca2+ current was tonically inhibited in neurons expressing the mutant K192A receptor, which has no affinity for anandamide, demonstrating that this receptor is also tonically active. SR 141716A had no effect on the Ca2+ current in these neurons, but SR 141716A could still antagonize the effect of WIN 55, 212-2. Thus, the K192 site is critical for the inverse agonist activity of SR 141716A. SR 141716A appeared to become a neutral antagonist at the K192A mutant receptor. Native cannabinoid receptors were studied in male rat major pelvic ganglion neurons, where it was found that WIN 55,212-2 inhibited and SR 141716A increased Ca2+ currents. Taken together, our results demonstrate that a population of native and cloned CB1 cannabinoid receptors can exist in a tonically active state that can be reversed by SR 141716A, which acts as an inverse agonist. PMID- 9855636 TI - Up- and down-regulation by glucocorticoids of the constitutive expression of the mast cell growth factor stem cell factor by human lung fibroblasts in culture. AB - Stem cell factor (SCF) is a major mast cell growth factor that promotes differentiation and chemotaxis of mast cells and inhibits their apoptosis. SCF therefore may be involved in diseases associated with an increased number of tissue mast cells such as asthma, for which the major treatment is glucocorticoids. In this study, we evaluated the effect of the glucocorticoid budesonide on the constitutive expression of SCF by human lung fibroblasts in primary culture. Budesonide (0.1 microM) induced a time-dependent biphasic effect on SCF mRNA and protein production. A short treatment (2.5-10 hr) induced an inhibition of SCF protein accumulation (-58% at 2.5 hr) and mRNA expression (-69% at 2.5 hr), associated with an accelerated decay of SCF mRNA and with a decrease in SCF gene transcription observed by nuclear run-on assay. Longer treatment (24 72 hr) led to increases in SCF protein accumulation (+64% at 48 hr) and mRNA expression (+125% at 24 hr) as a consequence of transcriptional activation. Similar effects of a decrease followed by an increase in SCF production were observed using another glucocorticoid, dexamethasone. Overall, our results show that glucocorticoids potently regulate SCF expression in human lung fibroblasts, successively decreasing and increasing SCF mRNA levels according to treatment duration. Such time-dependent modulation of SCF levels may explain some current discrepant findings about the effects of glucocorticoids on SCF production and may have functional consequences during glucocorticoid treatment, such as asthma therapy. PMID- 9855637 TI - Mibefradil inhibition of T-type calcium channels in cerebellar purkinje neurons. AB - The antihypertensive agent mibefradil completely and reversibly inhibited T-type calcium channels in freshly isolated rat cerebellar Purkinje neurons. The potency of mibefradil was increased at less hyperpolarized holding potentials, and the apparent affinity was correlated with the degree of channel inactivation. At 35 degrees, the apparent dissociation constant Kapp was 1 microM at a holding voltage of -110 mV (corresponding to noninactivated channels) and 83 nM at a holding voltage of -70 mV (corresponding to 65% inactivation). The increased affinity was attributable mainly to a decreased off-rate. Mibefradil also inhibited P-type calcium channels in Purkinje neurons, but inhibition was much less potent. At a holding potential of -70 mV, the Kapp for mibefradil inhibition of P-type channels was approximately 200-fold higher than that for inhibition of T-type channels. Mibefradil should be a useful compound for distinguishing T-type channels from high voltage-activated calcium channels in neurons studied in vitro. PMID- 9855638 TI - Chimeric receptor analysis of the ketanserin binding site in the human 5 Hydroxytryptamine1D receptor: importance of the second extracellular loop and fifth transmembrane domain in antagonist binding. AB - The 5-hydroxytryptamine (5-HT)1B/1D receptor subtypes are involved in the regulation of 5-HT release and have gained particular interest because of their apparent role in migraine. Although selective antagonists for both receptor subtypes recently have been developed, the receptor domains involved in the pharmacological specificity of these antagonists are defined poorly. This was investigated with a chimeric 5-HT1B/1D receptor analysis and using ketanserin as a selective antagonist of h5-HT1D (h5-HT1D) Ki = 24-27 nM) as opposed to h5-HT1B (Ki = 2193-2902 nM) receptors. A domain of the h5-HT1D receptor encompassing the second extracellular loop and the fifth transmembrane domain is necessary and sufficient to promote higher affinity binding (Ki = 65-115 nM) for ketanserin to the h5-HT1B receptor. The same domain of the h5-HT1B receptor, when exchanged in the h5-HT1D receptor, abolished high affinity binding of ketanserin (Ki = 364 1265 nM). A similar observation was made with the antagonist ritanserin and seems specific because besides the unmodified binding affinities for 5-HT and zolmitriptan, only minor modifications (2-4-fold) were observed for the agonists L 694247 and sumatriptan and the antagonists GR 127935 and SB 224289. Generating point mutations of divergent amino acids compared with the h5-HT1B receptor did not demonstrate a smaller peptide region related to a significant modification of ketanserin binding. The antagonists ketanserin and ritanserin are likely to bind the h5-HT1D receptor by its second extracellular loop, near the exofacial surface of the fifth transmembrane domain, or both. PMID- 9855639 TI - The benzodiazepine binding pocket of recombinant alpha1beta2gamma2 gamma aminobutyric acidA receptors: relative orientation of ligands and amino acid side chains. AB - Wild-type alpha1beta2gamma2 gamma-aminobutyric acid (GABA)A receptors and receptors containing a point-mutated subunit gamma2F77Y were expressed by transient transfection in human embryonic kidney 293 cells. Mutant receptors bound the benzodiazepine binding site ligand [3H]flumazenil with similar, subnanomolar affinity as wild-type receptor. Displacement studies with diazepam showed that the affinity for this compound was reduced 250-fold on mutation, indicating that the tyrosine hydroxyl group interferes with diazepam binding. This differential behavior then was used to find the chemical entity presumably interacting with the phenyalanine residue in position 77 of the gamma2 subunit of wild-type receptors. Thirty-four substances were analyzed in this respect. Our results suggest that the phenyl substituent of diazepam is located close to gammaF77. Similarly, we investigated the possible location of alpha1T206 and gamma2M130. Electrophysiological data obtained with the wild-type receptor furthermore suggest a simple overlap between positive allosteric modulators acting at the benzodiazepine binding site with its antagonists. PMID- 9855640 TI - Functional coupling of human L-type Ca2+ channels and angiotensin AT1A receptors coexpressed in xenopus laevis oocytes: involvement of the carboxyl-terminal Ca2+ sensors. AB - A human recombinant L-type Ca2+ channel (alpha1C,77) was coexpressed with the rat angiotensin AT1A receptor in Xenopus laevis oocytes. In oocytes expressing only alpha1C,77 channels, application of human angiotensin II (1-10 microM) did not affect the amplitude or kinetics of Ba2+ currents (IBa). In sharp contrast, in oocytes coexpressing alpha1C,77 channels and AT1A receptors, application of 1 nM to 1 microM angiotensin gradually and reversibly inhibited IBa, without significantly changing its kinetics. The inhibitory effect of angiotensin on IBa was abolished in oocytes that had been preincubated with losartan (an AT1A receptor antagonist) or thapsigargin or injected with 1,2-bis(o aminophenoxy)ethane-N,N,N',N'-tetraacetate, pertussis toxin, guanosine-5'-O-(2 thio)diphosphate, or heparin, suggesting that the recombinant alpha1C channels were regulated by angiotensin through G protein-coupled AT1A receptors via activation of the inositol trisphosphate-dependent intracellular Ca2+ release pathway. Consistent with this hypothesis, no cross-signaling occurred between the AT1A receptor and a splice variant of alpha1C lacking Ca2+ sensors (alpha1C,86). The data suggest that the regulation of recombinant L-type Ca2+ channels by angiotensin is mediated by inositol trisphosphate-induced intracellular Ca2+ release and occurs at the molecular motif responsible for the Ca2+-induced inactivation of the channels. PMID- 9855642 TI - Agonist action of adenosine triphosphates at the human P2Y1 receptor. AB - The agonist selectivity for adenosine di- and triphosphates was determined for the human P2Y1 receptor stably expressed in human 1321N1 astrocytoma cells and was studied under conditions in which nucleotide metabolism was both minimized and assessed. Cells were grown at low density on glass coverslips, encased in a flow-through chamber, and continuously superfused with medium, and Ca2+ responses to nucleotides were quantified. Superfusion with high performance liquid chromatographically purified ADP, ATP, 2-methylthio-ADP, and 2-methylthio-ATP resulted in rapid Ca2+ responses, with EC50 values of 10 +/- 5, 304 +/- 51, 2 +/- 1, and 116 +/- 50 nM, respectively. Similar peak responses were observed with maximal concentrations of these four agonists and with the hydrolysis-resistant adenine nucleoside triphosphate adenosine-5'-O-(3-thiotriphosphate). No conversion of [3H]ATP to [3H]ADP occurred under these conditions. Similar full agonist activities of ATP, 2-methylthio-ATP, and ADP were observed in human embryonic kidney 293 cells, which natively express the P2Y1 receptor. In contrast to these results, Leon et al. [FEBS Lett 403:26-30 (1997)] and Hechler et al. [Mol Pharmacol 53:727-733 (1998)] recently reported that, whereas ADP and 2 methylthio-ADP were agonists, ATP and 2-methylthio-ATP were weak antagonists in studies of the human P2Y1 receptor expressed in human Jurkat cells. To assess whether differences in the degree of receptor reserve might explain this discrepancy of results, P2Y1 receptor-expressing 1321N1 cells were incubated for 24 hr with adenosine-5'-O-(2-thiodiphosphate), with the goal of down-regulating the level of functional receptors. Pretreatment with adenosine-5'-O-(2 thiodiphosphate) resulted in a 10-fold rightward shift in the concentration effect curve for ADP; in contrast, the agonist activity of ATP was completely abolished. Taken together, our results indicate that adenosine di- and triphosphates are agonists at the human P2Y1 receptor. However, the intrinsic efficacy of ATP is less than that of ADP, and the capacity of ATP to activate second messenger responses through this receptor apparently depends on the degree of P2Y1 receptor reserve. PMID- 9855641 TI - Environmental xenobiotics and the antihormones cyproterone acetate and spironolactone use the nuclear hormone pregnenolone X receptor to activate the CYP3A23 hormone response element. AB - The pregnenolone X receptor (PXR), a new member of the nuclear hormone receptor superfamily, was recently demonstrated to mediate glucocorticoid agonist and antagonist activation of a hormone response element spaced by three nucleotides (DR-3) within the rat CYP3A23 promoter. Because many other steroids and xenobiotics can up-regulate CYP3A23 expression, we determined whether some of these other regulators used PXR to activate the CYP3A23 DR-3. Transient co transfection of LLC-PK1 cells with (CYP3A23)2-tk-CAT and mouse PXR demonstrated that the organochlorine pesticides transnonachlor and chlordane and the nonplanar polychlorinated biphenyls (PCBs) each induced the CYP3A23 DR-3 element, and this activation required PXR. Additionally, this study found that PXR is activated to induce (CYP3A23)2-tk-CAT by antihormones of several steroid classes including the antimineralocorticoid spironolactone and the antiandrogen cyproterone acetate. These studies reveal that PXR is involved in the induction of CYP3A23 by pharmacologically and structurally distinct steroids and xenobiotics. Moreover, PXR-mediated PCB activation of the (CYP3A23)2-tk-CAT may serve as a rapid assay for effects of nonplanar PCBs. PMID- 9855643 TI - Four pharmacologically distinct subtypes of alpha4beta2 nicotinic acetylcholine receptor expressed in Xenopus laevis oocytes. AB - Nicotinic receptors generally are presumed to consist of two alpha and three non alpha subunits. We varied the relative levels of expression of the neuronal nicotinic alpha4 and beta2 receptor subunits in Xenopus laevis oocytes by nuclear injection of cDNAs coding for these subunits in alpha:beta ratios of 9:1, 1:1, and 1:9. The sensitivities of the receptors to acetylcholine and d-tubocurarine were investigated in voltage-clamp experiments. For receptors expressed at the 9:1 and 1:1 alpha:beta ratios, the EC50 value of acetylcholine is approximately 60 microM. For the majority of the receptors expressed at the 1:9 alpha:beta ratio, the sensitivity to acetylcholine is enhanced 30-fold. No evidence for more than one type of acetylcholine binding site in a single receptor is obtained. The sensitivity to d-tubocurarine decreases with decreasing alpha:beta ratio. IC50 values of d-tubocurarine are 0.2, 0.5, and 2 microM for the 9:1, 1:1, and 1:9 alpha:beta ratios, respectively. At the 1:9 alpha:beta ratio, additional receptors with an IC50 value of 163 microM d-tubocurarine are expressed. At least two components with distinct sensitivities to d-tubocurarine are required to account for the shift in IC50. The combined agonist and antagonist effects reveal four distinct subtypes of alpha4beta2 nicotinic receptors. The results imply that the subunit stoichiometry of heteromeric alpha4beta2 acetylcholine receptors is not restricted to 2alpha:3beta. PMID- 9855645 TI - Cycloguanil and its parent compound proguanil demonstrate distinct activities against Plasmodium falciparum malaria parasites transformed with human dihydrofolate reductase. AB - The lack of suitable antimalarial agents to replace chloroquine and pyrimethamine/sulfadoxine threatens efforts to control the spread of drug resistant strains of the malaria parasite Plasmodium falciparum. Here we describe a transformation system, involving WR99210 selection of parasites transformed with either wild-type or methotrexate-resistant human dihydrofolate reductase (DHFR), that has application for the screening of P. falciparum-specific DHFR inhibitors that are active against drug-resistant parasites. Using this system, we have found that the prophylactic drug cycloguanil has a mode of pharmacological action distinct from the activity of its parent compound proguanil. Complementation assays demonstrate that cycloguanil acts specifically on P. falciparum DHFR and has no other significant target. The target of proguanil itself is separate from DHFR. We propose a strategy of combination chemotherapy incorporating the use of multiple parasite-specific inhibitors that act at the same molecular target and thereby maintain, in combination, their effectiveness against alternative forms of resistance that arise from different sets of point mutations in the target. This approach could be combined with traditional forms of combination chemotherapy in which two or more compounds are used against separate targets. PMID- 9855644 TI - Neuronal nicotinic receptor beta2 and beta4 subunits confer large differences in agonist binding affinity. AB - We used equilibrium binding analysis to characterize the agonist binding properties of six different rat neuronal nicotinic receptor subunit combinations expressed in Xenopus laevis oocytes. The alpha4beta2 receptor bound [3H]cytisine with a Kdapp of 0.74 +/- 0. 14 nM. The rank order of Kiapp values of additional nicotinic ligands, determined in competition assays, was cytisine < nicotine < acetylcholine < carbachol < curare. These pharmacological properties of alpha4beta2 expressed in oocytes are comparable to published values for the high affinity cytisine binding site in rat brain (alpha4beta2), demonstrating that rat neuronal nicotinic receptors expressed in X. laevis oocytes display appropriate pharmacological properties. Use of [3H]epibatidine allowed detailed characterization of multiple neuronal nicotinic receptor subunit combinations. Kdapp values for [3H]epibatidine binding were 10 pM for alpha2beta2, 87 pM for alpha2beta4, 14 pM for alpha3beta2, 300 pM for alpha3beta4, 30 pM for alpha4beta2, and 85 pM for alpha4beta4. Affinities for six additional agonists (acetylcholine, anabasine, cytisine, 1, 1-dimethyl-4-phenylpiperazinium, lobeline, and nicotine) were determined in competition assays. The beta2 containing receptors had consistently higher affinities for these agonists than did beta4-containing receptors. Particularly striking examples are the affinities displayed by alpha2beta2 and alpha2beta4, which differ in 1,1-dimethyl-4 phenylpiperazinium, nicotine, lobeline, and acetylcholine affinity by 120-, 86-, 85-, and 61-fold, respectively. Although smaller differences in affinity could be ascribed to different alpha subunits, the major factor in determining agonist affinity was the nature of the beta subunit. PMID- 9855646 TI - Vascular lesions of the spinal cord: history and classification. AB - Vascular lesions of the spinal cord are uncommon causes of myelopathy. Although recognized for more than 100 years, only slightly more than 100 cases were reported in the literature by midcentury. With recognition of the clinical syndrome and the development of selective spinal angiography in the 1960s and 1970s, a systemic classification of these diverse lesions has been made, allowing rational treatment using modern microsurgical and endovascular therapy. PMID- 9855647 TI - Vascular anatomy of the spinal cord. AB - Knowledge of spinal cord vascular anatomy is essential to the neurosurgeon planning and undertaking procedures on vascular lesions of the spinal cord. This article relates general concepts and nomenclature regarding this unique circulation and describes the regional and segmental arterial and venous anatomy of the spinal cord. The article concludes with surgical considerations and recommendations based on this anatomy. PMID- 9855648 TI - Clinical syndromes, natural history, and pathophysiology of vascular lesions of the spinal cord. AB - With superselective angiography and careful clinical evaluation, dural fistulas were found to be the most important and most frequent origin of spinal arteriovenous malformations. Diagnosis may not be made for months or years, however, because dural fistulas never bleed and clinical deterioration is slow. Therapy with interventional techniques or microsurgery is relatively safe. Perimedullary fistulas and intramedullary angiomas mostly deteriorate quickly because of hemodynamic or hemorrhagic problems. Close teamwork of interventionalists and neurosurgeons is mandatory. PMID- 9855649 TI - Endovascular treatment of spinal vascular malformations. AB - Endovascular treatment is considered as the first choice of treatment for most cases of spinal vascular malformations. Clinical presentation, natural history, and origin of symptoms are briefly described on spinal cord arteriovenous malformations, spinal dural arteriovenous fistulas, epidural fistulas, and paraspinal fistulas. Pretherapeutic evaluation, materials and technique for embolization, post-therapeutic management, results, and follow-up are discussed for each disease. PMID- 9855650 TI - The surgical management of spinal dural vascular malformations. AB - Spinal dural arteriovenous fistulas (AVFs) are the most common spinal vascular malformation. The spinal dural AVF is a disease of adults, primarily men, that is a cause of subacute progressive lower extremity myelopathy. The challenge for the optimum management of these patients is in making an accurate, early diagnosis. Definitive surgical intervention by interruption of the intradural draining vein is the treatment of choice. PMID- 9855651 TI - Spinal arteriovenous malformations. AB - Spinal cord arteriovenous malformations encompass a variety of distinct lesions with aberrant connections between the arterial and venous circulation. Symptomatology and clinical presentation of such lesions are directly related to their location and particular pathophysiology. In the authors' surgical experience, 90% of patients have had the same or improved neurologic function and long-term follow-up. PMID- 9855652 TI - Intramedullary cavernous malformations of the spinal cord. AB - Cavernous malformations of the spinal cord are extremely rare lesions. The 58 reported cases in the English literature and 9 of the authors' own cases are reviewed. The clinical presentation, patient characteristics, radiographic appearance, and histopathologic features are reviewed. The optimal surgical management and outcomes of treatment for intramedullary spinal cord cavernous malformations are discussed in detail. PMID- 9855653 TI - Outcome from treatment for spinal arteriovenous malformation. AB - Our knowledge of the outcome from treatment for spinal arteriovenous malformations (AVMs) is limited to a few small, anecdotal series and is thus far from complete. Furthermore, with the recent rapid evolution of our knowledge of AVM pathology, many reported cases are difficult to place in the current classification system. In general, outcome for the four classifications of AVMs are as follows: type I respond well to surgical disconnection of the arteriovenous fistula if treatment is early; type II AVMs are currently best treated by surgery but risks of a poor outcome need to be carefully considered; type III are only rarely treatable; type IVa and IVb are likely to respond well to surgery but our knowledge is limited; and type IVc are likely to fare poorly from surgery but should have a better outcome from endovascular therapy. PMID- 9855654 TI - Spinal vascular malformations: endovascular therapy AB - Over the past 30 years, the development of diagnostic spinal angiography has led to effective endovascular techniques for treatment of spinal vascular malformations. Specific locations, size, rapidity of shunt, and clinical presentation determine the best treatment and prognosis. PMID- 9855655 TI - Sudden painless visual loss: optic nerve and circulatory disturbances. AB - There are many causes of sudden painless visual loss in older adults. This article reviews the causes that are based on disturbances of the optic nerve and disorders of the circulatory system of the eye. Clinical clues to the recognition of these disorders and treatment (if available) are reviewed. PMID- 9855656 TI - Sudden painless visual loss: retinal causes. AB - Sudden painless visual loss occurs in retinal disorders that reflect primary ocular disease, as well as systemic disease. This article reviews vitreous and retinal detachment and macular degeneration as ocular causes of sudden visual loss. Retinal changes that are caused by systemic disease, including infectious retinitis, occlusion of retinal vessels, and proliferative new vessel formation, are reviewed. In each instance, the retinal examination should provide the ocular or systemic diagnosis or lead to the diagnosis. PMID- 9855657 TI - Gradual painless visual loss: retinal causes. AB - This article addresses retinal causes of gradual painless visual loss in older adults. The most common condition causing gradual vision loss is nonexudative age related macular degeneration, for which no effective treatment exists. Diabetic retinopathy is another common cause of gradual visual loss in older adults, in which photocoagulation, as well as vitreous surgery, may be helpful in preserving or restoring vision. Macular hole and macular pucker, which may cause central visual loss, are treatable with intraocular surgery, and visual improvement may be achieved in the majority of patients. Inflammatory causes of gradual visual loss in older adults are less common, and may benefit from immunosuppressive therapy. PMID- 9855658 TI - Gradual painless visual loss: chronic optic neuropathies. AB - The clinician must be the ultimate medical detective when dealing with chronic optic neuropathies. History taking is crucial. Clinical examination may require supplementation with visual field testing, fluorescein angiography, ocular and orbital ultrasound imaging, CT and MR imaging, blood test data, and cerebrospinal fluid or tissue biopsy data to determine the specific diagnosis. This supplementation is labor-intensive and time-consuming; the visual loss usually will progress throughout the process, frustrating and frightening the patient and physician. The final common pathway is gradual optic atrophy; the appearance of the optic nerve is rarely adequate to determine the cause of the visual loss. This article includes tables that review diagnostic aids and therapies, and lists the frequency with which several disease entities were encountered over 15 years in one tertiary care neuro-ophthalmic practice. If a specific cause is discernible, then a specific therapy may be available. This approach has the best chance of saving the patient's vision with the least toxicity caused by erroneous trials. By necessity, the work-up for these patients is expensive, but the cost of not pursuing the cause is irrevocable, permanent blindness. PMID- 9855659 TI - Gradual painless visual loss: anterior segment causes. AB - There are estimated to be 40 million blind individuals throughout the world. The majority of blind people in developing and developed countries are older adults. As the number of people over 60 years of age increases, the number of blind people also will increase. The primary care physician can play a major role in diagnosing and preventing many blinding diseases. This article discusses some of the major anterior segment causes of visual impairment in older adults in the industrialized nations. PMID- 9855660 TI - Gradual painless visual loss: glaucoma. AB - Glaucoma is one of the leading causes of blindness in adult Americans. This disease remains undetected in one half of the people with this disease. Primary open angle glaucoma is the most common form of the disease. Patients at risk for glaucoma usually have one or more of the following characteristics: advanced age, diabetes, myopia, elevated intraocular pressure, large cup to disc ratio, and a positive family history of the disease. Presently in the United States, treatment for primary open angle glaucoma is usually medical. Six classes of glaucoma medications are available, all of which have side effects. The second most prevalent form of glaucoma is angle closure glaucoma, the treatment for which is usually surgical. Although a cure for the various forms of glaucoma does not exist, an awareness of the problem and early detection and treatment minimize the impact of the disease. PMID- 9855661 TI - The painful eye: external and anterior segment causes. AB - When a patient presents to a medical practitioner with a painful eye, the initial history is extremely valuable in determining the cause of the complaint. The patient should be questioned specifically about the onset and duration of symptoms; description of the pain; exacerbating and mitigating factors; associated pruritus, discharge, or photophobia; and any previous occurrences. It is important to inquire about the patient's past medical history, past ocular history (including surgeries, trauma, contact lens wear, and family history of glaucoma), systemic and ocular medications, and allergies. A careful examination of the patient's skin, face, eyelids, conjunctiva, sclera, cornea, and anterior chamber should be performed. In this article, the authors describe a variety of external diseases and anterior segment causes of a painful eye, many of which can be diagnosed from the initial history. The article works systematically, beginning externally with the eyelids and conjunctiva and progressing internally toward the cornea and anterior chamber. PMID- 9855662 TI - The painful eye: neuro-ophthalmic causes. AB - A physician should have a better appreciation of diseases that can cause significant ocular morbidity and, in some cases, death. This article describes some of the painful eye syndromes associated with ocular, orbital, and intracranial disease. Differential diagnosis, clinical findings, laboratory investigation, and treatment are reviewed. PMID- 9855663 TI - Prevention of visual loss in older adults. AB - The major causes of visual loss in older adults are cataracts, glaucoma, age related macular degeneration, and diabetic retinopathy. Intensive research is ongoing for the prevention of these diseases. The present article updates different research studies and clinical trials aimed at prevention of visual loss in older adults. PMID- 9855664 TI - Hearing loss with aging: presbycusis. AB - Older adults with presbycusis have varied presentations. Early identification requires sensitivity to communication problems described by patients and their families. Early amplification may lead to better success in hearing aid use and may prevent social isolation. PMID- 9855665 TI - Nonsurgical management of hearing loss. AB - This article presents the salient points involved in age-induced hearing loss (i.e., presbycusis). The general evaluation, screening, diagnosis, and treatment modalities are discussed. Also, advances in hearing aid technology are described. PMID- 9855666 TI - Otolaryngologic management of dizziness in the older patient. AB - Studies report that dizziness is the most common presenting symptom in older patients who seek primary care. To understand the causes of dizziness, which is one of the major risk factors in causing falls in the older population, basic anatomy and physiology are reviewed in this article. Age-specific histopathologic changes occur in the labyrinth of the inner ear. To display evidence of the patient's symptoms, evaluation must include a comprehensive history, neurotologic examination, and diagnostic testing. There are common disorders associated with dizziness; thus, management depends on the cause. Once a diagnosis is secured, treatment is instituted based on sound medical principles. PMID- 9855667 TI - Tinnitus: etiology and management. AB - Tinnitus is the perceived sensation of sound in the absence of acoustic stimulation. Individuals who suffer from it are commonly between the ages of 40 and 80 years. Tinnitus is often classified as objective or subjective, yet the pathophysiologic cause is still unknown. Subjective tinnitus is largely identified with hearing loss. Management of tinnitus is based on an individual approach; there is no single treatment or regimen for it. PMID- 9855668 TI - Low back pain: current physiologic concepts. AB - It is clear that low back pain is not a simple process that can be explained on purely anatomic, biomechanical, or neurophysiologic theories. The intimate relationship of these processes have to be considered. Each of the potential biomechanical stresses influence the different spinal structures in different ways, depending on the forces applied and the structure being affected. The biomechanical response to the trauma caused by these forces may be immunologic, inflammatory, or neurochemical, depending on the structures being traumatized. Once a nociceptive process is underway, it is greatly influenced by the spinal cord and the brain. Only by following this process through its various steps can a clinician begin to formulate an understanding of the mechanisms involved in the genesis of low back pain. PMID- 9855669 TI - The clinical relevance of biomechanics. AB - Most neurologists are familiar with biomechanics but may be unsure of the relevance of this field to their practice. Actually those involved in musculoskeletal problems are undoubtedly using biomechanical principles. This article is limited to the spine, but the basic principles of biomechanics are applicable to other parts of the body. In this article, we describe the spine and trunk as a biomechanical organ, the biomechanical principles behind back injuries and their importance, the role of biomechanical issues in pain, the utility of clinical tests based on biomechanical principles, the effects of aging, and the future directions in spine biomechanical research. PMID- 9855670 TI - Differential diagnosis: a reasonable clinical approach. AB - The reasonable clinical approach to the patient who has low back pain includes a search for any of the red flags that would prompt consideration of additional diagnostic testing for serious underlying disease. The common patterns of back pain would be explored in an effort to categorize the patient's symptoms and to begin formulation of a diagnostic impression. Re-evaluation of the patient in light of his response (or lack of response) to these measures is an essential component of the clinical assessment. Finally, some consideration must be given to the various biopsychosocial factors that may affect prognosis, particularly in patients who have more chronic pain and disability-related issues. PMID- 9855671 TI - Clinical neurophysiologic studies: which test is useful and when? AB - Neurophysiological studies test the integrity of nerve roots and nerves at different points from the spinal cord to the periphery. EMG is the most sensitive neurophysiological test for evaluating patients with radiculopathy, providing information on diagnosis, location, and prognosis. F-wave and H-reflex studies may be abnormal, but the information that they provide is nonspecific and usually redundant because the needle examination is abnormal anyway. Somatosensory evoked potentials are less sensitive and specific than EMG for diagnosing a radiculopathy, but uncommonly may be the only abnormality. Magnetic stimulation, paraspinal mapping, and cervical root stimulation are investigational techniques of uncertain utility. PMID- 9855673 TI - Spinal manipulation: current state of research and its indications. AB - Based on the most recent and comprehensive systematic reviews, there is moderate evidence of short-term efficacy for spinal manipulation in the treatment of both acute and chronic low back pain. There is insufficient data available to draw conclusions regarding the efficacy for lumbar radiculopathy. The evidence is also not conclusive for the long-term efficacy of spinal manipulation for any type of low back pain. PMID- 9855672 TI - Physical therapy: exercises and the modalities: when, what, and Why? AB - This article reviews the evidence for using modalities and/or exercise treatment in patients with nonspecific low back pain. Poor evidence of efficacy exists for the use of modalities in this patient group. Exercises are beneficial for patients with subacute and chronic nonspecific low back pain. Further studies are needed for type, frequency, duration, and intensity of exercises. PMID- 9855674 TI - Surgery: indications and options. AB - The challenge is to better define the present role of spine surgery and more completely understand its realistic benefits and consequences. In spite of modern sophistication, spine surgery can still accomplish only the same two broad objectives, decompression and fusion, that have been possible for more than a century. Never vague or tentative, surgery has its place in the narrow spectrum of cases with clearly established, well circumscribed, structural pathology unresponsive to nonsurgical treatment and producing significant persistent disability. Identifying valid indications and options within the plethora of choices requires knowledge of what is available, what can actually be accomplished, and what limitations apply. PMID- 9855675 TI - Rehabilitation: what do we do with the chronic patient? AB - Neurologists are often called on to see patients who have low back pain presenting with significant chronicity and disabling pain. Even in situations of chronic low back pain, it has been estimated that a structural diagnosis is made only 60% of the time. Even when a physical diagnosis is made in these cases, it may be irrelevant to the primary causes of persistent pain and disability. This article is designed to point out that, when nonstructural factors are adequately rehabilitated, even in a worst-case occupational injury cohort, remarkable outcomes can be anticipated irrespective of the structural pathology, patient age, or postoperative impairment. PMID- 9855676 TI - Psychosocial issues: their importance in predicting disability, response to treatment, and search for compensation. AB - The conceptualization of pain and its progression into chronic disability has evolved from unidimensional models to more integrative, biopsychosocial models that take into account the many biological, psychosocial, social, and economic factors that may significantly contribute to the low back pain experience. This chapter reviews various studies that have demonstrated our growing understanding of these complex, interactive processes in helping to predict those who develop chronic disability as well as those who respond best to treatment attempts. Further, we examine the issue of compensation and how it too is intricately intertwined with the other variables contributing to lower back pain disability. PMID- 9855677 TI - Disability: how successful are we in determining disability? AB - Individuals who have severe back pain have trouble bending, may not be able to put on their shoes, have difficulty in ambulation, and may not be able to clean their own houses. What distinguishes these individuals with low back pain from those afflicted with other conditions (i.e., congestive heart failure, metastatic cancer, symptomatic acquired immune deficiency syndrome) is prognosis. Back pain essentially never shortens life. Acute back pain has a very benign prognosis, with more than 90% of the individuals returning to functional status equivalent to their baseline status within 3 months of the onset of pain. Patients who have chronic back pain have a significantly worse prognosis, but most cohort studies show that substantial numbers of chronic back pain patients improve over time with supportive therapy. PMID- 9855678 TI - Perils, pitfalls, and accomplishments of guidelines for treatment of back problems. AB - Science should be the basis for guidelines. As a result of the Flexner Report in 1911, we now live in an era where randomized trials are available. Statistical methods can truly be applied to evaluate the reliability of data published in the literature. The result is that we can now demand more from future publications and allow for a better evaluation of the mistakes or bias that can distort validity, applicability, and reliability. The importance of this methodology is to reduce misunderstandings by patients, clinicians, manufacturers, and government agencies about issues important to patient care. PMID- 9855679 TI - Poly(ADP-ribose) polymerase, a potential target for drugs: Cellular regulatory role of the polymer and the polymerase protein mediated by catalytic and macromolecular colligative actions (Review). AB - The cellular coenzymatic role of NAD, being a pleiotropic cofactor for diverse cellular reactions, is extended to poly(ADP-ribose) and to the highly abundant nuclear protein, poly(ADP-ribose) polymerase, with special focus on the pharmacological action of ligands on the latter. The polymer is defined to possess a helical configuration. From direct analyses of the polymer under physiological conditions, it is concluded that the polymerase is dormant in normal tissues, but is activated under certain pathological conditions: malignancy, retroviral integrate containing cells, and in a variety of inflammatory states. The interaction of poly(ADP-ribose) polymerase ligands with the DNA component of the active poly (ADP-ribose) polymerase - DNA complex is shown. A major cellular function of the poly(ADP-ribose) polymerase protein is its binding capacity to a large number of nuclear proteins and DNA sites, an effect which is induced by drugs that inhibit the polymerase activity. The malignancy-reverting effect of poly(ADP-ribose) polymerase ligand drugs is illustrated in chemically and oncovirally transformed cancer cells. The poly(ADP ribose) polymerase ligand-induced cessation of HIV replication is analyzed. Peroxynitrite-induced DNA damage-initiated pathological responses are shown to be inhibited by a specific poly(ADP-ribose) polymerase ligand. The irreversibly acting C-NO drugs oxidize asymmetric zinc fingers [poly(ADP-ribose) polymerase, HIV gag-precursor protein] and act as anti-cancer and anti-HIV agents, an effect that is regulated by cellular concentration of GSH. PMID- 9855680 TI - Mechanisms of restenosis after angioplasty and approach to therapy (Review). AB - Angioplasty is a principal treatment for occlusive vascular disease but 30-50% of patients show a restenosis of the vessel. There is no clinical effective therapy for this disease. It has been demonstrated, in animal models, that various drugs such as NO-donor, plasminogen inhibitor tranexamic acid and MMP (matrix metalloproteinases) reduce the rate of restenosis. Other therapeutic approaches are cytotoxic therapy, and strategies to inhibit cell cycle progression. Systemic administration of conventional pharmacologic agents inhibit cell cycle kinases and vascular lesion formation in animal models. As cell cycle progression is accompanied by fluctuations in the concentration of adenosine 3',5-monophospate (cAMP) and in the activity of the cAMP dependent protein kinase (PKA), local administration of cAMP and phospodiesterase-inhibitor drugs (aminophylline or amrinone) markedly inhibit neointima formation. The successful use of local radiation therapy to inhibit neointima formation after vascular injury may reflect a similar combination of cell-cycle arrest and vascular cell apoptosis. The most effective therapy for occlusive vascular disease will likely combine intravascular stenting with an antiproliferative therapy. PMID- 9855681 TI - Synthesis, biological effects and pathophysiological implications of the novel arachidonic acid metabolite 5-oxo-eicosatetraenoic acid (Review). AB - Arachidonic acid is rapidly metabolized by several distinct enzymes including the 5-lipoxygenase generating leukotrienes and 5-hydroxyeicosatetraenoic acid (5 HETE). These well studied metabolites cause a variety of physiological and pathophysiological effects in different tissues. Recently, oxidation of 5-HETE to 5-oxo-eicosatetraenoic acid (5-oxo-ETE) by an NADP+-dependent dehydrogenase has been demonstrated. Calcium ionophors and protein kinase C activators stimulate the synthesis of 5-oxo-ETE in neutrophils, eosinophils and monocytes. This novel arachidonic acid metabolite has a potent chemotactic activity for neutrophils and eosinophils. It stimulates adhesion of neutrophils and induces reactive oxygen metabolites in eosinophils. There is evidence that 5-oxo-ETE and 5-HETE interact with a specific G-protein coupled receptor. Since in contrast to 5-oxo-ETE much higher concentrations of 5-HETE are needed to provoke cell responses, 5-oxo-ETE might be the physiological relevant ligand for this putative receptor. Further downstream signalling pathways of this ligand include calcium transients, actin polymerization, activation of phosphatidylinositol-3-kinase and MAP-kinase. 5-oxo ETE has been extracted from scales of psoriatic patients and injection of 5-oxo ETE into rabbit subcutis causes a severe edema with an inflammatory cell infiltrate resembling an urticarial lesion. These findings indicate, that 5-oxo ETE might play a role in different cutaneous inflammatory diseases. PMID- 9855682 TI - Interleukin-2 at different concentrations induces the growth of selective lymphoid cells. AB - The effects of low and high doses of recombinant interleukin-2 (rIL-2) on cultured peripheral blood mononucleated cells are reported with the aim to show the effects of this immunomodulator in different conditions. The proliferation of various cell types at different IL-2 concentrations was investigated and ultrastructural and enzymatic studies were performed. The data obtained indicate that grade and type of cell stimulation induced by IL-2 is correlated to the dose employed. PMID- 9855683 TI - In situ localization of paclitaxel binding structures: Labeling with a paclitaxel fluorescent analogue. AB - Microtubules are a major component of cell cytoskeleton. Microtubules constitute the cellular target of paclitaxel. The interaction of paclitaxel with microtubules causes an increase in tubulin polymerization and microtubules stabilization, leading to a G2/M phase cell cycle arrest and cell death by apoptosis. Three paclitaxel fluorescent analogues were prepared by introducing fluorescein or BODIPY moiety onto the 2' of the 7-carbon. These were then used to study the interaction of paclitaxel with its cellular binding site and the microtubule network was visualized directly by fluorescence microscopy. Free paclitaxel was able to inhibit 7-FITC paclitaxel binding, demonstrating that both products bind to the same site and possess similar biological properties. Using the carbon 7 derivatives, microtubules were labeled as cytoplasmic fibers extending centripedly. A few other cellular components such as nuclear membrane, nucleoli and some other cytoplasmic structures were also labeled. The labeling intensity was reduced by preincubation with free paclitaxel. The interaction of paclitaxel with microtubules was also investigated using flow cytometry. No binding of the 2'C derivative was detected, confirming that a free 2'C is a pre requisite for paclitaxel microtubules interaction. PMID- 9855684 TI - Lack of B7.1 and B7.2 on head and neck cancer cells and possible significance for gene therapy. AB - Cellular immunity mediated by cytotoxic T-cells plays a key role in host tumor defense. An optimal T-cell recognition is based primarily on the presentation of an antigen in the context with MHC Class I molecules, secondarily co-stimulatory molecules, such as the B7 family, are necessary to initiate maximum stimulation. We examined the quality (immunohistochemically) and quantity (FACS-analysis) of B7 expression on primary and permanently established head and neck squamous cell cancer (HNSCC). Neither on the primary nor on the permanently established HNSCC lines could B7 be detected. The lack of co-stimulatory molecules could be the reason for the low immunogenicity of HNSCC. Future gene transfer of B7 could help to restore the immune function. Transfection of expression plasmids encoding B7 cDNA into the tumor cells by gene therapy might restore tumor-specific immunity as a new tool for cancer eradication in the future. PMID- 9855685 TI - Gain of DNA copy number on chromosomes 3q26-qter and 5p14-pter is a frequent finding in head and neck squamous cell carcinomas. AB - Comparative genomic hybridization (CGH) was used to study genomic imbalances in 77 squamous cell carcinomas of the head and neck (HNSCC) and in four cell lines derived from oral carcinomas. Particular attention was paid to all tumors characterized by a gain of two specific chromosomal segments, i.e. 3q26-qter and 5p14-p15. The 57 tumors containing both or one of the two imbalances were compared with 20 tumors lacking both with regard to genomic complexity, as well as to associated genomic, histopathologic and clinical peculiarities. 60% of the oral, and 66% of the non-oral cancers exhibited a gain of 3q26-qter, while a gain of 5p14-p15 was found in 66% of the oral, but only in 48% of the non-oral tumors. 48% of all tumors were characterized by both gains together, 26% exhibited only one of the two alterations. It could be shown that presence of both, gain of 3q26 qter and 5p14-p15, was clearly associated with a significantly higher complexity of genomic changes which was not only expressed as a high frequency of DNA copy number alterations (CNAs) but was also connected with a considerable number of additional amplifications of various chromosomal segments and a high conformity of the patterns of genomic imbalances in these tumors. Clear differences of the extent and of the patterns of genomic imbalance could be observed between oral and non-oral tumors. With respect to histopathological parameters, no clear association could be found for specific imbalances to the grade of differentiation, nor the invasiveness or metastatic status of the tumors. However, a higher number of patients with tumors characterized by gain of 3q26 qter plus 5p14-p15 died within a short period (i.e. less than 15 months) after excision of the tumor compared to the group without these imbalances. The implications of these findings are discussed from the oncogenetic and clinical aspects and in comparison with other reports. PMID- 9855686 TI - Interferon-c induces cathepsin B expression in a human macrophage-like cell line by increasing both transcription and mRNA stability. AB - We have demonstrated previously that treatment of the phorbol ester PMA-primed THP-1 human macrophage-like cells with interferon-c (IFN-c) in vitro induced time and dose-dependent increases in steady-state levels of cathepsin B (CB) mRNA. The present study was undertaken to investigate the mechanism of that increase. In vitro nuclear transcription (nuclear run-off) assays of CB gene expression were performed with purified nuclei from IFN-c-treated or untreated THP-1 cells. These assays showed transcription to be increased approximately three-fold by IFN-c in the PMA-primed THP-1 cells. Studies with a-amanitin indicated that the half-life of CB mRNA is prolonged after PMA and IFN-c treatment, by more than 90%. Therefore, the elevated CB mRNA level results from a combination of IFN-c-induced increase in the transcription rate of the CB gene and stabilization of the corresponding transcripts. The IFN-c-mediated increase in CB gene transcription and steady-state mRNA level was blocked by a-amanitin or cycloheximide, suggesting the involvement of RNA polymerase II and the requirement of de novo protein synthesis. PMID- 9855687 TI - Purification of pig heart benzylamine oxidase. AB - A benzylamine oxidase (E.C. 1.4.3.6) has been purified from pig heart. Western blot analysis showed that the enzyme cross-reacts with a polyclonal antibody raised against homogeneous, crystalline pig plasma benzylamine oxidase (BAO). A subunit molecular mass of 97 kDa obtained by SDS electrophoresis is identical to the plasma enzyme. The purification procedure consisted of sequential DEAE cellulose, DEAE-Sephadex, Con A-Sepharose, Sephadex G 200 and hydroxyapatite columns. The specific activity of the purified enzyme was 0.037 umol min-1mg-1 at 37uC and the Km for benzylamine was estimated to be 29 uM. The enzyme was inhibited by carbonyl reagents such as semicarbazide and a-aminoguanidine. Phenylhydrazine reacts mole to mole with the enzyme giving a peak at 425 nm. The copper content was 2 g-atom/mole of enzyme. PMID- 9855688 TI - Balanced affinity AT1/AT2 receptor nonpeptide binding site determinants on the AT1 angiotensin receptor. AB - Angiotensin II (Ang II), a potent modulator of cardiovascular homeostasis, mediates its effects through two Ang II receptor subtypes (AT1 and AT2) which share less than 30% amino acid homology and can be differentiated by specific nonpeptide ligands. We investigated the structural determinants of ligand binding to the AT1 receptor for L-163,017 which belongs to a newly developed class of nonpeptides that have equivalent affinities for AT1 and AT2 receptors. Nonpeptide binding affinities were determined in radioreceptor binding assays in membranes from COS cells transfected with wild-type and mutant receptor cDNAs. The amphibian AT receptor variant, xCM46, recognized L-163,017 with an affinity (IC50=8+/-2 nM) that was only 1.5-fold lower than for the rat AT1 receptor (IC50=5+/-1 nM) which is in striking contrast to the poor affinity of the amphibian xATa wild-type receptor (IC50 >50 uM). Analysis of single point rAT1 receptor mutants in which individual mammalian residues were replaced by the corresponding frog amino acids revealed significant overlap but also distinct differences in ligand binding interactions between dual receptor and subtype selective non-peptides. These data suggest that AT1-selective and dual receptor nonpeptides share overlapping but distinct binding pockets on the AT1 receptor. These findings may lead to improved therapeutics for the treatment of cardiovascular diseases involving both AT receptor subtypes. PMID- 9855689 TI - Enhancement of wound tissue expansion and angiogenesis by matrix-embedded fibroblast (dermagraft), a role of hepatocyte growth factor/scatter factor. AB - This study examined the effects of matrix-embedded human fibroblasts, a predominant cell type in the injured tissue, on the tissue expansion and angiogenesis. Using a co-culture technique, it was demonstrated that the presence of matrix-embedded fibroblasts (Dermagraft) significantly enhanced the expansion of human wound tissue in a 3D gel system over a period of 10 days. Using a rat aorta ring assay, fibroblasts also significantly stimulated the growth of new vessels from the ring and also enhanced the motility of human vascular endothelial cells. This effect of fibroblasts was neutralised with anti-HGF/SF antibody. HGF/SF protein was detected in both supernatant and cell lysate of the fibroblasts by bioassay and Western blotting, mRNA for HGF/SF was detected in the fibroblasts by RT-PCR. HGF/SF secreted by the fibroblast was able to stimulate the phosphorylation of cMET, HGF/SF receptor. It is thus concluded that matrix embedded fibroblasts are capable of stimulating wound healing and this effect is attributed to HGF/SF, produced by the cell. PMID- 9855690 TI - 8-(3-oxo-4,5,6-trihydroxy-3h-xanthen-9-yl)-1-naphthoic acid inhibits MAPK phosphorylation in endothelial cells induced by VEGF and bFGF. AB - Angiogenesis is regulated by various factors. In particular, VEGF and basic FGF are of much importance. We found that 8-(3-oxo-4,5,6-trihydroxy-3h-xanthen-9-yl) 1-naphthoic acid inhibited the binding of VEGF to KDR/Flk-1 (VEGF receptor-2) or Flt-1 (VEGF receptor-1) and that it inhibited the MAPK phosphorylation in HUVEC induced by VEGF or basic FGF but not by EGF. 8-(3-oxo-4,5,6-trihydroxy-3h-xanthen 9-yl)-1-naphthoic acid might be used as an inhibitor of VEGF and basic FGF signal transduction. PMID- 9855691 TI - Comparison of nuclear matrix proteins between EGFR-antisense transfected and untransfected glioblastoma cells. AB - The protein composition of the nuclear matrix is both tissue and cell type specific, and it undergoes changes with differentiation and transformation. In the present study, nuclear matrix proteins of EGFR-antisense transfected glioblastoma cell lines, U87 and U343, were compared with untransfected cell lines using two dimensional-gel electrophoresis. After EGFR-antisense transfection, the protein compositions of the nuclear matrices in both cell lines were different. Several nuclear proteins were only found in EGFR-antisense transfected cell lines. There was no difference in NuMA expression in the transfected and untransfected cell lines. These results suggest that EGFR antisense reduced tumorigenicity on human glioblastoma cells by changing nuclear matrix protein compositions. PMID- 9855692 TI - Loss of heterozygosity and microsatellite instability in non-neoplastic mucosa from patients with chronic ulcerative colitis. AB - Microsatellite instability and allelic deletions of tumor suppressor genes have been observed frequently in tumors. Molecular pathogenesis of the development of dysplasia and carcinoma in ulcerative colitis is still unclear. In order to detect microsatellite alterations in ulcerative colitis, we analyzed loss of heterozygosity (LOH) and microsatellite instability (MI) on chromosomes 3, 6, 7, 12, and tumor suppressor gene loci, including p53, APC, and p16, of chronically inflamed, non-dysplastic epithelium after microdissection. Twelve of 13 (92%) cases showed LOH and/or MI at one or more loci. LOH at chromosome 3 and MI at chromosome 12 were observed in 50% and 62%, respectively. However, LOH at p53 and p16 was detected in only one case each. These results suggest that chronic inflammation may initiate microsatellite alteration, which subsequently transform ulcerative colitis to dysplasia or cancer. This finding provides information for the evaluation and treatment of patients with ulcerative colitis. PMID- 9855693 TI - The effects of a soybean extract on tumor growth and metastasis. AB - In vitro genistein (soybean-derived isoflavone/phytoestrogen) has been reported to suppress angiogenesis. We have produced a soybean extract (SE) enriched in phytoestogens which are heat stable. Heat sterilization (hs) of SE destroys soy protease inhibitor activity and increases isoflavone concentrations. This study was performed to determine the effects of SE with and without heat sterilization on tumor growth and metastasis. Sixty female Lewis rats injected s.c. with mammary tumor (MAC-33) were randomized to receive i.p. injection of SE (18 mg), hsSE (18 mg), or saline (control) 5 times per week for 30 days. After 30 days the rats were sacrificed to determine carcass weight, tumor weight, tumor volume, liver weight, hepatic protease activity (HPA), and number and size of lung metastasis. Comparing either SE or hsSE to controls, significant increases were found in tumor weight (g) (109.4+/-26.3, 114.9+/-19.7, 86.8+/-23.2), tumor volume (cm3) (55.6+/-14.6, 60.1+/-12.0, 45.5+/-14.4), and tumor: carcass ratio (0.59, 0.62, 0.47). Data expressed as mean +/- SD (SE, hsSE, control, respectively) was analyzed by ANOVA. There was a significant increase in the number of lung metastasis (31%; Kruskal-Wallis test) in the animals receiving hsSE. Additionally, there was a significant decrease in HPA (26.9%) in the animals receiving SE. Soy protease inhibitors are not responsible for the increase in tumor growth and number of lung metastases seen with SE. Our results suggest that a heat stable component of SE, perhaps the isoflavones, promotes tumor growth and lung metastasis in vivo. PMID- 9855694 TI - Estrogen receptors and glucocorticoid receptors in human well-differentiated thyroid cancer. AB - Previously, we and others have found that the prognosis of patients with well differentiated thyroid cancer is related to patient age and gender. In this study, we examined estrogen and glucocorticoid receptors in thyroid cancers and adjacent non-neoplastic thyroid tissues from 31 Chinese patients. Our results indicate that the level of estrogen and glucocorticoid receptors in thyroid cancer tissues was similar to that in non-neoplastic thyroid tissues. Significant higher estrogen and glucocorticoid receptor levels were found in thyroid cancers from men than women. There was no difference in estrogen receptor levels from thyroid cancer in women less than or older than 45. However, the level of glucocorticoid receptor in thyroid cancer from women less than 45 years old was significantly higher than that of women older than 45. Estrogen or glucocorticoid receptor levels did not correlate with tumor invasiveness. The pathobiological role of glucocorticoid receptors in thyroid cancer is not known but may be involved in changes in circadian rhythm of thyroid tumors. PMID- 9855695 TI - The role of natriuretic peptides in cardiovascular disorders and human cardiac allografts (Review). AB - The human myocardium is composed of a variety of cardiac cell types that synthesise cardioactive factors with autocrine, paracrine and endocrine functions. These cardioactive factors include a family of structurally and functionally related peptides, natriuretic peptides (NPs), that act as cardiovascular cell growth modulatory factors and have significant influence on the regulation of cardiovascular function. The three members of the NP family are atrial natriuretic peptide, brain natriuretic peptide and C-type natriuretic peptide. NPs are ubiquitously expressed in cardiovascular tissues and have specific receptors in cardiac cells, brain cells, vascular endothelial cells and extracardiac tissues through which they elicit a variety of biological responses including natriuresis, diuresis and vasodilation. Cardiac transplantation is the most effective and definitive treatment for end-stage cardiac failure in humans. Orthotopic cardiac transplantation and cardiovascular disorders including hypertension, cardiac hypertrophy, cardiac failure, result in increased myocardial expression of NPs, suggesting a pathophysiological role for these peptides in the heart. The mechanism by which the expression of NPs is regulated in the transplanted human heart is not well understood. Understanding the mechanism of local expression of NPs and their interactions with other putative growth regulatory factors in the human heart may have important implications for potential management of cardiovascular disorders and cardiac transplantation. This article will discuss the current knowledge of NPs in cardiovascular disorders. Most of the emphasis will focus on their possible role in human cardiac allografts as there have been no reviews on this important topic. PMID- 9855697 TI - Trafficking pathway of GLUT4 glucose transporters in muscle (review). AB - Skeletal muscle is a major glucose-utilizing tissue in the absorptive state and alterations in muscle insulin-stimulated glucose uptake lead to derangements in whole body glucose disposal. The major glucose transporter expressed in skeletal muscle is the GLUT4 isoform. In the muscle fiber, GLUT4 undergoes insulin stimulated translocation to T-tubules and to sarcolemma and it represents a pharmacological target for the treatment of diabetes mellitus, insulin-resistant states or in cardiac dysfunction. We review recent studies describing the characterization of the cellular steps followed by GLUT4 in muscle and we propose several working models for an integrated structure of the GLUT4 trafficking pathway. PMID- 9855696 TI - Carbon monoxide and vascular cell function (review). AB - Carbon monoxide (CO) is an endogenously generated gas that may play an important physiological role in the circulation. CO is generated by vascular cells as a byproduct of heme catabolism, in which heme oxygenase (HO) catalyzes the degradation of heme to biliverdin, iron and CO. Two distinct isoforms of HO have been identified in vascular tissue. The HO-2 isoform is constitutively expressed and likely mediates the release of CO under normal physiologic conditions. In contrast, the HO-1 isoform is strongly induced in vascular cells by various stress-associated agents and markedly increases CO synthesis during pathological conditions. The release of CO by vascular cells exerts both paracrine and autocrine effects on vascular smooth muscle cells (SMC) and circulating blood cells. CO regulates blood flow and blood fluidity by inhibiting vasomotor tone, SMC proliferation, and platelet aggregation. These vascular effects of CO are mediated via the activation of soluble guanylate cyclase and the consequent rise in intracellular guanosine 3',5'-cyclic monophosphate levels in target tissues. CO may also play a role in various cardiovascular disorders, including endotoxin shock, ischemia-reperfusion, hypertension, and subarachnoid hemorrhage. This review will focus on the recent progress made in understanding the regulation and function of CO in the vasculature. PMID- 9855698 TI - Regulation of apoptosis during development: input from the extracellular matrix (review). AB - Programmed cell death or apoptosis is an important aspect in organogenesis and tissue remodeling during development. Extensive investigations have led to the identification of many genes that participate in the regulation of cell death execution. These include the caspases and nucleases, which are involved in the degradation of cellular proteins and nuclear DNA to initiate the irreversible death process. In addition, several families of proteins like Bcl-2 superfamily can either prevent or promote cell death. The function of these proteins are getting to be understood. On the other hand, how these proteins are regulated remains to be investigated. This is in part due to the presence of diverse upstream signals that can influence cell fate. One such signal is the remodeling of the extracellular matrix (ECM), which is largely due to the action of matrix metalloproteinases (MMPs). The regulation of MMPs and ECM remodeling has been shown to affect apoptosis in different systems, including the apoptotic remodeling of the intestine during Xenopus laevis metamorphosis and post lactation involution of the mouse mammary gland. Current evidence suggests that ECM regulates cell fate at least in part through its membrane receptors, the integrins, which in turn send the signal through yet poorly understood pathways to the nucleus to regulate gene expression. PMID- 9855699 TI - Carboxyflavins, novel inhibitors of Taq DNA polymerase. AB - Carboxyflavins were found to be potent selective inhibitors of Taq DNA polymerase in a polymerase chain reaction. The inhibitions were dose-dependent, and complete inhibitions were observed at the concentration of 3.0 microM. Carboxyflavins were much less, or not sensitive to the DNA polymerases tested such as calf thymus DNA polymerase alpha, rat DNA polymerase beta, human immunodeficiency virus type 1 reverse transcriptase, the Klenow Fragment of E. coli DNA polymerase I and T4 DNA polymerase. To our knowledge, there is no other report of an agent that selectively inhibits only a thermophilic polymerase. Interestingly, the carboxyflavins were able to prevent DNA synthesis in the murine lymphoid leukemia cell line L1210 in vitro; almost complete inhibitory levels were achieved in the range of less than 10 microM. PMID- 9855700 TI - Clinical significance and molecular mechanisms of bioinactive growth hormone (review). AB - About 80% of short children are not deficient in endogenous growth hormone (GH) and termed idiopathic short stature (ISS). The causes of impaired growth in children with ISS are various. Short stature and low insulin-like growth factor-I (IGF-I) concentration despite normal to high GH concentration suggest impaired GH effect. The prototypical GH insensitivity syndrome was described and characterized by the absent or defective GH receptors. Growth retardation resulting from biologically inactive GH was also described, but the molecular basis of biologically inactive GH has remained unclear. Recently, two unique point mutations in the GH-1 gene in the children with short stature whose GH were supposed as bioinactive were reported. Mutant GH R77C not only failed to stimulate tyrosine phosphorylation by itself, but it also inhibited the activity of wild-type GH. This mutant GH exerted an antagonistic effect. Another mutant D112G was only bio-inactive. This case was a typical Kowarski syndrome. The molecular heterogeneity of mutant GH reflected clinical phenotype of bioinactive GH syndrome. PMID- 9855701 TI - Ryanodine receptors and their role in genetic diseases (review). AB - The skeletal muscle ryanodine receptor (RYR1) is a calcium release channel that mediates efflux of calcium ions from the sarcoplasmic reticulum into the myoplasm during excitation-contraction coupling. Mutations in the RYR1 gene have been detected in about 50% of the patients suffering from malignant hyperthermia (MH), but evidence is accumulating that other genetic defects can also lead to MH in humans. MH is a life-threatening disorder induced by exposure to volatile anesthetics and/or the muscle relaxans succinylcholin during surgical procedures in affected patients. MH leads to skeletal muscle rigidity, hypermetabolism and rapid rise in body temperature. MH is also known in pigs where it is triggered by stress and therefore often referred to as porcine stress syndrome. The existence of an animal model has greatly faciliated the elucidation of the basis for the human disease. This review describes recent advances in the understanding of the physiological action of ryanodine receptors and new insights regarding the relation between different RYR1 mutations and distinct phenotypical appearances. PMID- 9855702 TI - Changes in telomerase activity of advanced cancers of oral cavity and oropharynx during radiation therapy: correlation with clinical outcome. AB - Telomerase activity has been evaluated for many kinds of malignancies, and it has been clarified that the activity reflected the malignant potential of the tumor. With regard to radiation therapy for cancer, simple and reliable assays for the prediction of therapeutic effect have not been fully developed yet. Therefore, we aimed to determine whether telomerase activity is changed by radiotherapy for advanced head and neck malignancies, and to clarify the possible correlation of telomerase activity with the therapeutic effect of radiation therapy and patients' clinical outcomes. Twenty-five patients with advanced cancers of the oral cavity and oropharynx were examined. All the cancers were confirmed histopathologically as squamous cell carcinomas. Biopsies were performed before treatment and at doses of 4 and/or 10 Gy, and 20 Gy of radiotherapy. To semi quantitate telomerase activity, a highly sensitive PCR-based telomeric repeat amplification protocol assay was performed. Nineteen of the 21 cancers (90.1%) showed telomerase activity of varied extents prior to radiation therapy. Nine of the 25 patients showed continuous elevations of telomerase activity (> 10 TPG units/microg protein) in their tumors during radiation therapy, and these nine showed 13.8-216.9 TPG units/microg protein at 20 Gy of radiotherapy. In eight of the nine patients, tumors did not respond well to radiotherapy and relapsed locally in a short period after the treatment. On the other hand, in another group of 11 patients, who showed low (< 10) and/or no activity of telomerase in their tumors at 20 Gy of radiotherapy, there were nine patients whose tumors responded well to radiotherapy (p < 0.025 by the chi2 test). None of them relapsed locally in the follow-up period of approximately 21 months, and the difference was statistically significant (p < 0.025 by the chi2 test). It is concluded that the high activity of telomerase in tumor tissue at 20 Gy of radiotherapy can predict a poor therapeutic effect of radiation therapy and unfavorable patients' outcomes for advanced cancers of oral cavity and oropharynx. PMID- 9855703 TI - Quantitative analysis of gastrin mRNA and peptide in normal and cancerous human pancreas. AB - Gastrin has been shown to stimulate growth of human pancreatic cancer, and does so in an autocrine fashion. In this study, a relationship between gastrin mRNA, peptide, and gastrin receptors were studied in a variety of human pancreatic tissues. Low levels of gastrin mRNA were detected in normal human pancreas by quantitative reverse transcription polymerase chain reaction, but gastrin peptide was not present using radioimmunoassay. Pancreatic adenocarcinoma cells and tissues had 34- to 530-fold higher gastrin mRNA and peptide levels than normal pancreas. Gastrin mRNA and peptide levels were 8,000- and 15,000-fold, respectively, greater in a pancreatic islet cell gastrinoma tumor than in normal pancreas. In comparison to age-matched controls, fasting gastrin plasma levels were 2-fold higher in patients with pancreatic adenocarcinoma and 131-fold greater in subjects with gastrinomas. Receptor binding assays revealed that pancreatic cancer cells had a binding capacity 200-fold greater than gastrinoma tumors, and 10-fold greater than normal pancreas; no differences in K(d) values were recorded between specimens. In contrast to the normal pancreas and gastrinoma tumor, the aggressive behavior of pancreatic adenocarcinoma may be attributed to the autocrine production of gastrin and to the presence of its growth-related receptor. PMID- 9855704 TI - Optimal erythropoietin expression in human hepatoma cell lines requires activation of multiple signalling pathways. AB - Hypoxia is thought to be a common precursor of coronary artery disease and malignant tumors, both diseases representing the leading causes of death in industrial nations. So far, investigations of oxygen-regulated erythropoietin (EPO) gene expression in the human hepatoma cell lines Hep3B and HepG2 allowed many important insights into the mechanisms of oxygen-sensing, signalling and regulation of an increasing number of oxygen-responsive genes. To differentiate the various signalling pathways involved in EPO production by these two cell lines, we examined several factors that positively influenced EPO expression. The results demonstrate a keen differential effect of cell density and oxygen concentration on EPO induction in Hep3B compared to HepG2 cells. Using optimized cell culture conditions, EPO production rates as high as 1 U EPO per 10(6) Hep3B cells in 24 h could be achieved. We also found a moderate but reproducible positive effect of CoCl2 on hypoxia-induced EPO expression in Hep3B but a negative CoCl2 effect on hypoxic induction in HepG2 cells. CoCl2 inhibited cell growth in a concentration-dependent manner. Interleukin-6 was synergistic with hypoxia on EPO induction in Hep3B as well as HepG2 cells, and dexamethasone enhanced this effect in Hep3B but not in HepG2 cells. The moderate CoCl2 dependent increase of EPO production observed in hypoxic Hep3B cells might indicate that CoCl2 and hypoxia do not necessarily act via, identical signalling pathways. PMID- 9855705 TI - Suppression of HIV replication by dominant negative mutants of HIV-1 (review). AB - Various gag mutants of human immunodeficiency virus type 1 (HIV-1) generated in vitro were evaluated for their potentials to suppress the replication of wild type (wt) virus. A single-round of wt virus replication in the presence of various mutant proteins was quantitatively monitored by transfection and infection experiments. Out of 38 mutants examined, 15 were demonstrated to interfere with the replication of wt HIV-1 at early and/or late viral replication phase. Some of these mutants were also effective against the replication of wt HIV-2. In this review, we focus on the mutants, which are able to act against a wide variety of HIV, and are very useful for future gene therapy against AIDS. PMID- 9855706 TI - Inhibition of insulin release by alpha- and beta-D-galactose pentaacetate. AB - Several monosaccharide polyacetate esters display higher biological efficiency than the corresponding unesterified carbohydrates, this being attributable to their capacity to cross the plasma membrane without requiring the intervention of a specific carrier system and to their subsequent intracellular esterase catalyzed hydrolysis. Some of these esters, however, also exert a direct effect upon a receptor system apparently displaying analogies with that involved in the identification of bitter compounds by taste buds. For instance, under suitable experimental conditions, esters of non-nutrient monosaccharides, such as L glucose, D-mannoheptulose or 2-deoxy-D-glucose, unexpectedly stimulate insulin release. The present work reveals that alpha-D-galactose pentaacetate and, to a lesser extent, beta-D-galactose pentaacetate both inhibit leucine-induced insulin release in rat pancreatic islets. This indicates that the postulated receptor system displays anomeric specificity and may participate in a negative modulation of insulin secretion. It is proposed that advantage could be taken of the negative insulinotropic action of D-galactose pentaacetate esters to prevent excessive insulin release in pathological situations. PMID- 9855707 TI - Expression of human aldehyde dehydrogenase-3 associated with hepatocellular carcinoma: promoter regions and nuclear protein factors related to the expression. AB - The human ALDH3 gene is constitutively expressed in stomach, lung, esophagus, and cornea, but hardly detectable in the normal liver. However, it is highly activated in the hepatocellular carcinoma tissues from approximately 50% of patients. The nuclear DNA binding factors exist in both ALDH3-positive cancerous liver and ALDH3-positive HepG2 cells, but not in ALDH3-negative Hep3B cells and normal liver tissues. South-western blot hybridization showed the existence of two nuclear-binding protein components, 35 and 14 kDa, in ALDH3-positive cancerous liver tissues. These two DNA binding proteins were not found in normal stomach tissues and stomach carcinoma KATO III cells. DNaseI footprint analysis identified two protective regions within the ALDH3 promoter. The first protected region has one putative CCAAT-box and one putative Sp1-site. The second protected region contains a putative HiNF-A binding sequence. These findings suggest that a high level of expression of ALDH3 in cancerous liver tissues resulted from the expression or activation of at least two nuclear proteins reacting to the ALDH3 promoter region. PMID- 9855708 TI - The value of PCR technique in fine needle aspiration biopsy of salivary gland for diagnosis of low-grade B-cell lymphoma. AB - In fine needle aspiration biopsy (FNAB) of salivary gland delineation of low grade B-cell lymphoma from benign lymphoid lesions of myoepithelial sialadenitis (MESA) may be very difficult by means of cytomorphological criteria alone. To improve cytodiagnosis PCR technique was applied on routinely stained smears to determine clonal status by amplifying the third complementarity-determining region (CDR3) of the hypervariable domain of the immunoglobulin heavy chain. Twelve cases diagnosed cytologically as suspicious of low-grade B-NHL with following histology of B-NHL (n = 5) or MESA (n = 7) were analyzed. The CDR3-IgH PCR produced distinct bands in 10/12 cases. The PCR products were analyzed with Genescan software on the DNA sequencer, which demonstrated monoclonal bands in all NHLs and in one case of MESA. The results indicate that PCR technique may be helpful in improving cytodiagnostic accuracy for recognition of low-grade B-NHL of salivary gland. PMID- 9855709 TI - Comparative clinical study of cefonicid, chloramphenicol, and penicillin in community-acquired pneumonia. AB - Community-acquired pneumonia is one of the most common infectious conditions that require hospitalization. When intravenous treatment is indicated, cefonicid is usually the drug of choice. The aim of this study was to find out if chloramphenicol, which is superior to the standard drugs from a financial point of view, could serve as an equally efficient treatment, especially in the elderly. The outcomes of 3 pneumonia patient groups who were either treated with cefonicid, chloramphenicol or penicillin-G (n = 59, 17, 24, respectively) were retrospectively compared. Data about demographic characteristics of the patients, clinical outcomes, rehospitalization rates, duration of improvement/treatment/ hospitalization and clinical laboratory tests were obtained from each patient's medical records. Only minor differences (even though occasionally significant) were found with respect to rehospitalization and improvement rates, duration of hospitalization, treatment and improvement, death rates and clinical laboratory tests. However, chloramphenicol patients were found to be significantly older than cefonicid patients. Moreover, no bone-marrow suppression was associated with chloramphenicol treatment. All 3 drugs tested seem to have the same efficacy. We conclude that since chloramphenicol is as safe as, and much cheaper than cefonicid, this antibiotic agent is not inferior to the others, its usage in older patients with pneumonia should be considered. PMID- 9855710 TI - Increased expression of human histocompatibility leukocyte antigen-G in colorectal cancer cells. AB - Human histocompatibility leukocyte antigen (HLA)-G is a nonclassical major histocompatibility complex class I molecule. HLA-G is known to provide tolerance from recognition by natural killer cells. We studied HLA-G expression in 39 human colorectal cancers and 23 extra-neoplastic colon tissue samples by RT-PCR. The expression of HLA-G mRNA was significantly more frequent in colorectal cancer (34 of 39 cases) than in the extraneoplastic tissue (10 of 23 specimens; chi2 test, p = 0.0003). HLA-G expression was also confirmed on the cancer cells immunohistochemically. These results suggested that HLA-G on colorectal cancer cells may be correlated with escape from immunological surveillance during colon cancer development. PMID- 9855711 TI - 5'-flanking motifs control cell-specific expression of trefoil factor genes (TFF). AB - A group of secreted peptides (trefoil factor family; TFF) is abundantly expressed at mucosal surfaces of the gastrointestinal tract and promote epithelial restitution. They are upregulated around areas of epithelial damage, ulceration and neoplasia. The transcriptional regulation of the three human TFF genes was assayed by multiplex RT-PCR and reporter gene analysis in 8 gastrointestinal carcinoma cell lines. The level of endogenous mRNA matched well the reporter gene activity of all TFFs, indicating that the cis-acting elements located less than 1,000 bp upstream of the TATAA box account for cell-specific gene expression. In HT-29, the endogenous TFF expression profile changed in relation to cell growth conditions. Deletion and mutation analysis of TFF promoter constructs revealed enhancing elements shared within the three TFF promoters that were shown to bind nuclear proteins. Thus such specific DNA-protein interaction may explain the TFF peptides' cell specific expression pattern and altered levels in pathological conditions. PMID- 9855712 TI - Towards a change in status of emergency contraception? PMID- 9855713 TI - A review of bacteriological culture of removed intrauterine contraceptive devices. AB - A retrospective review of clinical and microbiological data was made, over an 18 month period, of intrauterine contraceptive devices removed and sent for examination. An association with symptoms at time of removal was found for Streptococcus agalactiae, Streptococcus milleri, enterobacteria and obligate anaerobes. Thirty four per cent of IUDs yielded one or more Actinomycesspecies spp but no correlation with symptoms was found. In the small group of symptomatic patients who were followed up, no difference in outcome was observed between those who were treated with antibiotics and those who were not. PMID- 9855714 TI - Contraceptive knowledge: development of a valid measure and survey of pill users and general practioners. AB - AIMS: To design, pilot, and validate a questionnaire to test contraceptive knowledge in combined oral contraceptive pill users. METHOD: To ensure face and content validity the questionnaire was developed using existing unvalidated instruments and in consultation with GPs, local and national family planning experts, and with pill users. The questionnaire was then piloted with 15 current oral contraceptive pill users and 10 local GPs, modified, and a contraceptive knowledge 'score' developed. Construct validity-that the degree of family planning training should predict contraceptive knowledge-was tested in four groups: family planning trainers (n=28), GPs (n=40), current pill users (n=53), and male medical students (n=59). Thirty current pill users were sent the questionnaire after two weeks to determine test-retest reliability. RESULTS: The questionnaire showed construct validity: there was a gradient of scores across the four knowledge groups. Family planning trainers had the highest scores, followed by GPs, and current pill users, with male medical students having the lowest scores (Kruskal-Wallis test p<0.001). Women had good knowledge of situations when pill efficacy is reduced but poorer knowledge of what action to take subsequently. Predictors of knowledge in pill users were educational level, age, and the importance attached to not falling pregnant-thus providing further evidence for the construct validity of the questionnaire. Test-retest reliability was good (rank correlation 0. 73). CONCLUSION: A contraceptive knowledge questionnaire suitable for use in audit or research has been developed which is reliable and has face, content and construct validity. Pill users have poor knowledge of what to do in situations of pill failure and about the details of emergency contraception. Trials are needed to assess the effectiveness of different strategies to improve contraceptive knowledge in women. PMID- 9855715 TI - Attenders at young people's clinics in Southampton: variations in contraceptive use. AB - This paper presents findings from a survey of 424 people attending nine young people's clinics within the Southampton Community Health NHS Trust. In addition to recording some descriptive background data on the people attending the clinics, one major aim of the study was to investigate whether talking to the sexual partner about contraception before their first intercourse together and delaying this first intercourse influenced contraceptive use. Overall, 40 per cent of people attending the clinics were aged 16 or under, although there was some variation between clinics in the age groups attracted. Most clients were female (88 per cent), had ever had sexual intercourse (92 per cent), reported four or more lifetime partners (42 per cent) but only one partner within the last six months (73 per cent) and were currently in a relationship (75 per cent). Potential for contraception and sexually transmitted infection was widespread; 46 per cent (of non-virgins) had had intercourse without contraception at least 'a few times' and 18 per cent used condoms 'rarely' or 'never'. In terms of first intercourse with current/most recent partner, 17 per cent had not used any contraception and 32 per cent had failed to use condoms. The most important findings from this study were that use of contraception (and condoms in particular) on the occasion of first intercourse with the current or most recent partner was significantly associated with the following; if partners had talked to each other about contraception before having intercourse together for the first time (p<0.001), and also if this first intercourse was delayed beyond four weeks as opposed to over a few days of first 'going-out' together (p<0.001). Suggestions for further in-depth research are made. PMID- 9855716 TI - Patient satisfaction with a direct access vasectomy service. AB - This study of patient satisfaction with a direct access vasectomy service was carried out by questionnaire offered to 100 patients attending a day surgery unit for vasectomy. Seventy eight patients responded and there was overall satisfaction with the service, with 72 patients (96 per cent of respondents) describing it as excellent or good. The commonest criticism expressed by respondents concerned long waiting times in the unit before surgery on the day of operation. Vasectomy is a straightforward and minor surgical procedure which suits a direct access service. PMID- 9855717 TI - Factors determining the choice of contraceptive methods at the Family Planning Clinic, University College Hospital, Ibadan, Nigeria. AB - In a study of 2000 women volunteers seeking contraceptive services at the Family Planning Clinic (FPC), University College Hospital, Ibadan, Nigeria, 66.2 per cent chose the intrauterine device (IUD) making it the most common method of contraception. Factors influencing choice of contraceptive methods were advice from friends and family members, intended duration of use and information from the media. Ignorance, fear and unfounded cultural beliefs were factors responsible for the delay in seeking contraceptive advice. The mass media was an important source of information for most of the women. We conclude that the IUD is the contraceptive of choice in our clinic because of the highly selective nature of our clients. In order to provide a service with a broader clientele, we suggest the incorporation of other priority reproductive health services such as cervical and breast cancer screening, prevention and treatment of reproductive tract infections and sexually transmitted diseases including HIV/AIDS. PMID- 9855718 TI - IUD training in non specialist centres. PMID- 9855719 TI - Norplant. PMID- 9855720 TI - [Xenotransplantation, soon a clinical reality? Biological barriers must be overcome]. PMID- 9855721 TI - [Preventive work against HIV/AIDS has been successful in Sweden. But the global transmission is still unchanged]. PMID- 9855722 TI - [Pardon me, but are violent men only a social phenomenon?]. PMID- 9855723 TI - [Physician's tasks are essential in health care]. PMID- 9855724 TI - [Handling of duplicate publications]. PMID- 9855725 TI - [Are animal cells and organs solution for the transplantation problem?]. PMID- 9855728 TI - [A new strategy in the fight against HIV/AIDS is needed. Inaccurate information is a waste of both money and energy]. PMID- 9855729 TI - [National and international guidelines ignore significant cause of heart failure. Diastolic dysfunction is often the cause of heart failure in the elderly and in women]. AB - Comparison of patients treated for chronic heart failure at a large hospital with patients included in major treatment studies published during the past ten years yielded important information. The former series was characterised by greater proportions of the elderly and of women than were the series recruited to the often cited ACE (angiotensin-converting enzyme) inhibitor studies. Although only patients with systolic dysfunction were recruited to the latter studies, a substantial body of evidence suggests the prevalence of severe heart failure among patients with normal systolic function to increase with increasing age. Thus, as many as 50 per cent of all elderly patients with chronic heart failure may have normal systolic function. In most cases, the heart failure is probably due to diastolic dysfunction, a condition that still lacks both a simple diagnostic procedure and a well-documented treatment. PMID- 9855730 TI - [Study and treatment of heart failure require new approaches. Natriuretic peptides can be safe and simple diagnostic and outcome measures]. AB - Analysis of plasma natriuretic peptides and related propeptide fragments may be a cost-effective aid to diagnostic evaluation and treatment follow-up in cases of heart failure. In diagnostic potential such variables may constitute first-line measures of high negative predictive value, allowing further examination, e.g. by echocardiography, in cases where values are above the respective cut-off levels. However, in many cases evaluation of published reports is rendered difficult by their omission of information on such pre-analytical variables as blood sampling and storage, and drug therapy. Moreover, different analytical methods may yield widely divergent results. Thus, before such assays are introduced in general use, their long-term validity needs to be ensured, for instance by consistency in calibration, and measurements need to be made in representative series of unselected patients for the determination of appropriate cut-off levels. PMID- 9855731 TI - [High risk for children with DAMP/ADHD to become addicts later in life]. AB - The prevalence of attention deficit hyperactivity disorder (ADHD) and of the overlapping condition, deficit in attention, motor control and perception (DAMP), among children of early school age is approximately 5 per cent (1.5% being severe cases). Boys are more commonly affected than girls. The symptoms continue to be disabling at 20 years of age in 50 per cent of cases, and social maladjustment is common. Half of the young boys develop oppositional defiant disorder (ODD), very often progressing to conduct disorder (CD) and antisocial personality disorder (ASPD). Low socio-economic status, parental mental disorder, and persistence of DAMP/ADHD symptoms are all predictors of the development of CD/ASPD. Prospective studies of children with DAMP/ADHD have shown them to be characterised by a high level of alcohol and/or drug abuse comorbidity, particularly the subgroup progressing to CD/ASPD. Conversely, studies of drug abuse and of alcohol abuse series have shown them to be characterised by a high level of DAMP/ADHD comorbidity. Approximately one in five alcoholics has or has had DAMP/ADHD, a comorbidity probably even more pronounced among type II alcoholics. Approximately every third substance-abuser has or has had DAMP/ADHD. The need of more persevering support and treatment efforts for the well-defined category of boys at high risk is emphasised. Better awareness, active diagnosis and treatment of adults with persisting DAMP/ADHD is also warranted. PMID- 9855732 TI - [A survey of "heavy consumers" of emergency care services in Stockholm. Six per cent of the patients accounted for a quarter of emergency consultations]. AB - A small subgroup of patients accounts for a substantial proportion of emergency room (ER) consultations. Of the 479,956 consultations at ER facilities at Stockholm hospitals in 1996, a quarter were accounted for by six per cent of the patients, a subgroup characterised by at least four ER consultations during the year. Some 1,500 patients consulted 12-233 times. Analysis of the care-seeking habits of five extremely heavy ER-facility consumers showed them also to be heavy consumers of other health care services such as primary and tertiary facilities. Since previous studies have shown heavy ER-facility consumers to be a vulnerable subgroup both medically and socially, co-ordination and planning of their care might well be beneficial. PMID- 9855733 TI - [Early treatment can improve prognosis in diabetic nephropathy]. AB - Prognosis in diabetic nephropathy has changed dramatically during the past decade, and slowing of the disease process has been made possible by intervention against specific risk factors. Nonetheless, diabetic nephropathy has become the leading cause of end stage renal disease. Early detection and prevention, or at least delaying, of disease progression have become crucial aims, and several treatment strategies designed to prevent end stage renal disease have recently been published. The common denominator of these strategies is screening for microalbuminuria in diabetic patients rather than awaiting the appearance of overt symptoms. PMID- 9855734 TI - [What hygienic criteria should apply to community care? The risk of infection is increasing when more and more seriously ill patients are treated outside hospital]. AB - In Sweden, the provision of health care (i.e. non-physician care) of elderly residents in community housing and care facilities has been the statutory responsibility of local authorities since 1992. The Health and Medical Services Act stipulates that such care should be of good quality, As infection control is one aspect of good quality, in 1997 the Dept. of Infection Control at University Hospital, Uppsala, and the six municipalities of the County of Uppsala that are responsible for the care of a total of 4,400 elderly people, launched a joint enterprise to provide the care required by the Act. The program includes the education of staff in the basic concepts of infection control/recruitment of link surses and surveillance of risk factors, in order to minimise nosocomial infections in nursing homes for the elderly. PMID- 9855735 TI - [Pheochromocytoma. The risk to miss diagnosis of severe disease is reduced when free metanephrines are measured in plasma]. PMID- 9855736 TI - [Vulvar complaints in young girls need instant professional evaluation]. AB - As vulvovaginal disease are nog very common in prepubescent girls, it may seem reasonable to suspect sexual abuse when a small girl presents with complaints in the genital area. However, the very fact that there are visible changes suggests that it is no a case of sexual abuse, the signs of which are usually more subtle. To arrive at a correct diagnosis, and avoid unnecessary, traumatic investigation into the possibility of sexual abuse, it is essential that such patients be examined from the outset by physicians experienced in this field. PMID- 9855737 TI - [No genetic counseling prior to the marriage of the future Czar]. PMID- 9855738 TI - [Chrysotile asbestos in Russia: certain results and promising research directions]. AB - The article summarizes studies carried in RAMSc Research Institute for Occupational Medicine on chrysotile asbestos. Not denying potential carcinogenicity characteristic for all kinds of asbestos, those studies stress low biologic aggression of chrysotile asbestos during occupational exposure, even if the excessive MAC is demonstrated formerly in asbestos industry enterprises. Work with chrysotile asbestos, as every one in mining industry, requires not ban, but accomplishment of proper measures aimed to prevent pneumoconiosis and dust bronchitis. The article demonstrates unique experience of Russian scientists- evaluation of exposure to chrysotile without admixtures and amphibole additives. The authors define prospective research trends that, if being international, could correctly solve problems associated with further use of chrysotile asbestos, rising no "anti-asbestos" boom. PMID- 9855739 TI - [Sanitary evaluation of microclimate conditions at the ore mining and alluvial pits sites]. AB - The authors summarized results of hygienic research carried in pits of Transbaikal region, Yakutia. Tchukotka, Kazakhstan, Kirgizia, Krivoy Rog, European North and other areas. Microclimate parameters in winze of mining and alluvial pits appeared to vary significantly and to depend on certain specific factors (depth of mining, climate geographic zone of the enterprise, temperature of the ores treated, etc.). Considering degree of influence on workers' heat state, taking into account significance of clothes heat insulation required, the authors defined 4 types of microclimate at workplace. For operative selection of optimal special clothing set, the authors represent dependence of proper heat resistance of clothes for main and auxiliary occupations on microclimate conditions. PMID- 9855740 TI - [Immunological disorders in vibration-related illness among miners of Northern regions]. AB - Evaluation of miners in Northern and East Siberian regions revealed disorders of humoral immunity: increased B-lymphocytes count, higher production of autologic antibodies to peripheral nerves, greater serum concentration of circulating immune complexes and cryoglobulins. Cytostatic medications suppressed antibodies production and considerably increased efficiency of complex therapy of severe vibration disease. PMID- 9855741 TI - [The influence of superoxide dismutase and catalase on the course of the pathological process in the lungs in experimental silicosis]. AB - The authors studied role of antioxidant enzymes--superoxide dismutase and catalase--in pathogenesis of experimental pulmonary silicosis in rats. Experimental silicosis was induced by intratracheal administration of quartz dust, 20 mg. Aerosols of superoxide dismutase and catalase water solutions were inhaled twice a week during first month and 5 times a week over the next 1.5 months. The results perfectly match the previous data proving quartz dust to stimulate granulocytes' release of extracellular superoxide radicals and hydrogen peroxide. The research results also demonstrate important role of agents moving one electron for reduction of oxygen in silicosis pathogenesis. PMID- 9855742 TI - [The body's hormonal balance in miners at the coal mines]. AB - The hormonal background of coal-pit miners was studied by radioimmunoassay method. It was determined, that adaptation to coal-dust influence proceeds by protective-compensatory increasing of cortisol, thyroid hormones and insulin levels. When a functional reserves of endocrine glands exhausted, the adaptation breaks down and pneumoconiosis may arise. PMID- 9855743 TI - [Correctional aspects of anisometropia in railway machine operators]. AB - The authors assessed occupationally significant visual functions in anisometropy varying in type and degree. Disorders of occupationally significant visual functions depend reliably on anisometropy degree. Correction of anisometropy with recovered vision and binocular functions restores all parameters of night and color vision. Optic and operative correction of anisometropy could be used for rehabilitation of railway machine operators with long length of service. The article provides recommendations to use intraocular lenses for artiphakia in railway machine operators. PMID- 9855744 TI - [The main pathogenetic mechanisms in occupational diseases of lungs of dust related etiology. Part I. From an experiment to a concept]. PMID- 9855745 TI - [The meaning of age- and sex-related aspects of the higher nervous system function for the mastering of operator occupations by teenagers]. PMID- 9855746 TI - [The 75th anniversary of the Russian Academy of Medical Sciences-based Occupational Medicine Institute]. PMID- 9855747 TI - [Obituary to Vladislav Iakovlevich Shustov]. PMID- 9855748 TI - Physicians' billing errors and omissions insurance. PMID- 9855750 TI - MSMS adopts "model" harassment policy. PMID- 9855749 TI - Physician integration options. You're not cats, they're not ranchers. PMID- 9855751 TI - Ethics in managed care. A matter of focus, a matter of integrity. PMID- 9855752 TI - The spiritual basis of medical practice. PMID- 9855753 TI - Medicare + Choice. PMID- 9855754 TI - The place of spirituality and religion in mental health services. AB - Mental health professionals have raised concerns about the role of spirituality and religion in services for people with severe mental disorders, but this chapter offers compelling reasons for increased attention to spiritual issues in service delivery. PMID- 9855755 TI - Assessment of spirituality and implications for service planning. AB - Assessing the role that religious or spiritual concerns play in the lives of people with severe mental disorders is crucial if clinicians are to determine whether those concerns can be successfully incorporated into service planning. PMID- 9855756 TI - Recoiling, regrouping, and recovering: first-person accounts of the role of spirituality in the course of serious mental illness. AB - Because many consumers view spirituality as important in their recovery, mental health professionals can benefit from a clearer understanding of the role of religion from the consumer's perspective. PMID- 9855757 TI - Spiritual and religious dimensions of mental illness recovery narratives. AB - Key themes in narratives of recovery from mental illness illustrate some of the functions of religion and spirituality in consumers' self-understanding and coping. PMID- 9855758 TI - Religious-issues group therapy. AB - The long-standing success of a Spiritual Beliefs and Values Group supports the potential value of discussing spiritual issues and challenges the assumption that religious concerns voiced by people with serious mental illness should be considered pathological. PMID- 9855759 TI - Spiritual dimensions of a mind-body group for people with severe mental illness. AB - In a mind-body group designed to address issues of well-being for people with severe mental disorders, experiences with spiritual themes of optimal functioning and ultimate meaning emerged with surprising clarity. PMID- 9855760 TI - The faith community as a support for people with mental illness. AB - Innovative programs developed by faith communities for people with mental illness, as well as guidelines for collaboration between religious and mental health organizations, are presented. PMID- 9855761 TI - Research on religion and serious mental illness. AB - According to this review, religion plays a largely positive role in mental health; future research on severe mental disorders should include religious factors more directly. PMID- 9855762 TI - Recommendations for integrating spirituality in mental health services. AB - On theoretical, clinical, and research levels, there are many ways to attend more fully to spirituality and religion in developing and implementing mental health programs. PMID- 9855764 TI - [Balneotherapy: the main results of the research of the last decade]. AB - The research data are presented obtained on hydrobalneotherapy effectiveness in cardiovascular, endocrine, bronchopulmonary, skin and combined diseases as well as effects of hydrobalneotherapy on meteosensitivity of cardiovascular patients. PMID- 9855765 TI - [The effect of mineral water on serotonin and insulin production (an experimental study)]. AB - Radioimmunoassay (DRG kits) and orthotoluidine test were conducted to measure blood serotonin, insulin and glucose in 70 intact Wistar rat males before and after a course of drinking mineral water Essentuki 17 (MW). After the MW drinking course, a single dose of mineral water increases basal levels of serotonin and insulin, sensitivity of endocrine cells to MW. Serotonin and insulin rose maximally on minute 5 after the drink while in contrast to minute 15 and 30 before initiation of the MW drinking course. A direct correlation was found between blood concentrations of serotonin and insulin. PMID- 9855766 TI - [The use of dry-air radon baths in the treatment of patients with reactive arthritis]. AB - 66 patients with reactive arthritis took dry air radon baths. Radon concentration was 1.5 kBq/l (40 nCi/l), temperature regimens 27 +/- 3 and 37 +/- 3 degrees C. The procedures produced analgetic, anti-inflammatory and immunocorrective effects. PMID- 9855767 TI - [The efficacy of psychorelaxation therapy in the rehabilitation of patients who have had an aortocoronary bypass operation]. AB - Psychorelaxation therapy based on the biological feedback was given to 43 patients after coronary artery bypass operation. 45 patients after this operation served control. One-month and one-year follow-up showed that patients of the study group had positive changes in the psychological status, they became less depressed and neurotic. Those who completed the course of psychorelaxation therapy exhibited higher performance and quality of life. PMID- 9855768 TI - [Bronchial asthma: new trends in the use of physical factors]. PMID- 9855769 TI - [An intensive regimen of pelotherapy in pulmonology]. AB - The authors show that intensification of peloid therapy, i.e. more frequent procedures, in patients with pulmonary diseases imposes additional load on the regulatory systems: vegetative, cardiovascular, immune. Moderate balneological reactions become more frequent, therapeutic results worsen. How to correct the above negative effects is proposed. PMID- 9855770 TI - [The immunomodulating action of low-energy laser radiation in the treatment of bronchial asthma]. PMID- 9855771 TI - [A clinico-thermographic assessment of the efficacy of naphthalan therapy in patients with diabetic microangiopathies of the lower extremities]. AB - Naphthalan was applied in 58 patients with diabetic microangiopathy of the lower limbs. Clinical and infra-red imaging proved high effectiveness of the treatment as naphthalan relieved symptoms of diabetic microangiopathy by means of correction of microcirculation and peripheral blood flow. Infra-red imaging showed that exercise and single procedure tests are able to give prognosis of the treatment effect. The success of naphthalan therapy is very important as microangiopathy of the lower limbs underlies serious complications in diabetics. PMID- 9855772 TI - [The characteristics of the mechanism of action in the joint use of solutions of brine and mud extracts with galvanization and ultrasonic exposure. 2]. AB - Radionuclide tracing has demonstrated that intraperitoneal introduction of natural brine and mud solutions to intact rats followed by ultrasound versus galvanization provides more intensive delivery of chemical elements to the target organs and tissues. These differences should be taken into account when planning rehabilitation schemes with various physiotherapeutic modalities. PMID- 9855773 TI - [The combined action of infrared radiation and permanent and alternating magnetic fields in experimental atherosclerosis]. AB - Paravertebral exposure to infrared radiation (0.87 micron, 5 mW) and permanent magnetic field in combination with one- and two-semiperiodic alternative magnetic fields (50 Hz, 15-30 mT) was studied in respect to the action on adaptive reactions in animals with experimental atherosclerosis. Complex consisting of infrared radiation, permanent magnetic field and one-semiperiodic pulse alternative magnetic field was most effective in restoration of vasomotor metabolic and immune disturbances accompanying development of atherosclerosis. PMID- 9855774 TI - [The use of physiotherapy and antibacterial preparations in urogenital infections]. AB - Antibacterial antibiotics only, antibiotics in combination with immunostimulators, antibiotics plus immunostimulators plus physiotherapy were administered in 132 patients with urethroprostatitis caused by mixed infection (18, 45 and 69 patients, respectively). A complete recovery was achieved only in patients who had been treated with the antibiotics, immunostimulators and physiotherapy. After antibiotics, residual infection was registered in 11.1% of the patients, clinical symptoms persisted in 6 patients; after antibiotics combined with immunostimulators in 6.7% and 4 patients, respectively. PMID- 9855775 TI - [The combined treatment of patients with chronic nonspecific salpingo-oophoritis using a low-frequency magnetic field and iodobromine water]. AB - The studies of 116 females with chronic nonspecific salpingo-oophoritis demonstrate that combination of low-frequency magnetic field with iodine-bromine water effectively corrects immunological indices, i.e. improves T- and B-cell immunity, production of IgM and IgA, proportion of immunoregulatory subpopulations of T-lymphocytes. PMID- 9855776 TI - [An attempt at typing the medicinal mineral waters of the Maritime Territory]. PMID- 9855777 TI - [Peloid preparations as a means of enhancing the efficacy of pelotherapy. I. The physicochemical characteristics of the organic substances in sulfide sludges]. PMID- 9855779 TI - [The efficacy of using millimeter-range electromagnetic radiation in chronic nonspecific pneumonia]. PMID- 9855778 TI - [The antimutagenic effect of gumizol in cultures of blood T-lymphocytes exposed to cyclophosphane]. PMID- 9855780 TI - [Physiotherapy in the rehabilitation of patients with chronic gastroduodenitis]. PMID- 9855782 TI - Health in Europe 1997. Report on the third evaluation of progress towards health for all in the European Region of WHO (1996-1997). PMID- 9855781 TI - [Experience in conducting topical lecture series for physician advanced training in puncture physiotherapy]. PMID- 9855783 TI - Organ donation: a matter of family. PMID- 9855784 TI - HIPPA law on Medicare home health care plans causes confusion. PMID- 9855785 TI - Shabby sales tactics or shameful acceptance of the goods? PMID- 9855786 TI - What's up with diabetes? PMID- 9855787 TI - When time is of the essence. PMID- 9855788 TI - Moratorium on transplant law pleases Wisconsin experts. PMID- 9855789 TI - Wisconsin donor registry proposal. AB - An opportunity exists to utilize an existing publicly supported information system as an economical means to address a significant health care issue. The creation of the Wisconsin Donor Registry will provide timely access to documentation regarding an individual resident's declaration of intent to be considered an organ or tissue donor. The Wisconsin Donor Registry will be the nations first legally binding registry of potential organ and tissue donors. This partnership between the state and the transplant community will ensure that the individual's right to direct this aspect of their health care is honored, that fewer opportunities for organ and tissue recovery are missed and, most importantly, a greater number of Wisconsin residents gain access to this scarce and valuable resource. PMID- 9855790 TI - Coronary stenting for coronary artery disease in a cardiac transplant recipient. AB - Accelerated coronary artery disease in the transplanted heart remains the leading cause of death in heart transplant recipients. Traditional treatment modalities have generally yielded sub-optimal results. Coronary artery stents are used frequently in the non-transplanted heart to treat coronary artery disease. Only a few cases using this approach in the cardiac transplant recipient have been reported. This report details the use of this modality in a transplant recipient with significant two-vessel coronary artery disease 11 years after orthotopic cardiac transplantation. PMID- 9855791 TI - Early experience with laparoscopic splenectomy. AB - BACKGROUND: Laparoscopic splenectomy is an example of the recent continued advancement in laparoscopic surgery as techniques are adapted for procedures previously done only via a laparotomy. METHODS: We analyzed our initial experience with laparoscopic splenectomy for details of the operative procedure and the clinical outcome including length of stay and complication rates. RESULTS: Two surgeons performed 11 laparoscopic splenectomies for cancer or hematologic disorders. In two instances (18%) conversion to open splenectomy was necessary due to bleeding at the splenic hilum. There were no mortalities. Two patients developed pancreatic fluid collections that were successfully drained percutaneously. The seven patients who had an uncomplicated course resumed eating a regular diet in 2.0 +/- 0.6 days (mu +/- SD) and had a hospital stay of 2.7 +/- 1.1 days. Hospital stay was significantly longer for the patients who had complications 9.7 +/- 7.2 days (p < 0.05) and for the 11 patients undergoing on elective uncomplicated open splenectomy during the same time period, 6.5 +/- 2.0 days (p < 0.05). For the 9 patients who had a completed laparoscopic splenectomy, the mean operative blood loss was 263.9 +/- 241.4 cc. The mean operative time was 293.3 +/- 91.4 minutes. The spleens removed laparoscopically weighed an average of 390 grams (range 17 to 1584 grams). CONCLUSIONS: Laparoscopic splenectomy can be performed safely and is associated with a rapid resumption of oral alimentation and shortened hospital stay compared to open splenectomy. Complications experienced in our early experience included conversion to open splenectomy and fluid collections from pancreatic leakage. PMID- 9855792 TI - Implantable atrial defibrillator for atrial fibrillation. AB - Atrial fibrillation is a common problem. Pharmacological and nonpharmacological approaches have limited therapeutic efficacy in many patients. Low energy transvenous atrial defibrillation using catheters positioned inside the right atrium and cornonary sinus has been shown to be efficacious in converting atrial fibrillation to sinus rhythm. We report the successful use of this technology in an implantable form in a patient with symptomatic atrial fibrillation. The patient could be kept in sinus rhythm using an implanted atrial defibrillator. PMID- 9855793 TI - Shotgun wound management: a comparison of slug and pellet injuries. AB - Shotgun slug injuries have received little attention while shotgun pellet wounds have been well described. Twenty-two shotgun pellet and 13 shotgun slug injuries treated over a 14-year period were retrospectively reviewed. Extremity and thoracic wounds were most frequent in both groups. The incidence of vascular and nerve injuries was similar for slug and pellet wounds. Angiography was more often used to evaluate pellet wounds for vascular disruption. The rate of wound infection was 38% for slug wounds versus 32% for pellet injuries. Tissue grafting was more frequently necessary for reconstruction after pellet injury. Long-term disability was documented in 15% of patients with pellet wounds and 23% with slug wounds. Despite similarities in wound location and outcomes, the ballistic differences between shotgun slugs and pellets resulted in significant differences in wounding characteristics and extent of injury which have important ramifications in management. PMID- 9855795 TI - Choosing annuities. PMID- 9855794 TI - Whatever happened to professional courtesy? The illegality of waiving co-pays and deductibles. PMID- 9855796 TI - Effect of root canal filling material and level of surgical injury on periodontal healing in dogs. AB - Some divergencies in the literature about periodontal healing after surgical injury stimulated the development of this experiment. The root canals of dogs' teeth were negotiated and filled by the lateral condensation technique with two kinds of sealers: Sealapex and zinc oxide-eugenol cement. In the second session, the bone tissue was exposed and one cavity was made at the apical third of the root and another at the border between the coronal and middle thirds, both penetrating into the root canal. Six months later the animals were sacrificed and the specimens prepared for histopathologic analysis. The results showed that the kind of filling material and the level of the periodontal wound exposing the root canal can influence the healing process. PMID- 9855797 TI - Initial treatment of traumatic dental injuries by dental practitioners. AB - The aim of this study was to investigate the nature of initial treatment provided by dental practitioners to children aged 1-17 years with various types of traumatic dental injuries at public dental clinics in Tanzania. Questionnaires on initial treatment were mailed to 188 practitioners and returned by 138 (73%). The reported treatments were analyzed in relation to the dental practitioners' qualifications and area of practice. Extraction of injured teeth was frequently reported for 64% of the injuries and prescription of antibiotics was reported by 67%, 48% and 46% of the practitioners for soft tissue injuries, concussion, and alveolar fracture respectively. Practitioners working at the Faculty of Dentistry were less involved in treating dental trauma than those at urban and rural clinics (P = 0.001), while no significant association was found with the level of education of the practitioners. Equal proportions of practitioners, about one third each, reported correct, unnecessary and wrong treatment options. The quality of the treatments provided could not be explained by background variables. It can be concluded that dental practitioners in Tanzania provide a lot of over-treatment for traumatic dental injuries. Therefore, it is suggested that efforts should be made to improve and standardize treatment methods in Tanzania. PMID- 9855798 TI - Influence of zinc oxide and eugenol sealer on concentration of zinc, calcium and copper in rat tissues. AB - A zinc oxide and eugenol root canal sealer (Roth 811) and sterile saline solution were injected into the dorsal thoracic midline of 70 male Wistar-Furth rats. Every day for the next 7 days, 10 animals were sacrificed by either inhalation. The liver, heart, kidneys and brain were removed from the animals and analysed for zinc, calcium and copper concentrations by flame atomic absorption spectrophotometry. The tissue around the injection site was also surgically removed and prepared for histological evaluation under a microscope. The injection of Roth 811 significantly affected the concentrations of zinc, calcium and copper in some of the examined organs, especially on the 4th and 5th day. The inflammatory reaction adjacent to the material was severe during the first 3 days while on the 7th day the presence of connective tissue with collagen formation was observed. PMID- 9855799 TI - Development of a computer-assisted learning software package on dental traumatology. AB - The development of computer-assisted learning software packages is a relatively new field of computer application. The progress made in personal computer technology toward more user-friendly operating systems has stimulated the academic community to develop computer-assisted learning for pre- and postgraduate students. The ability of computers to combine audio and visual data in an interactive form provides a powerful educational tool. The purpose of this study was to develop and evaluate a computer-assisted learning package on dental traumatology. This program contains background information on the diagnosis, classification, and management of dental injuries in both the permanent and the deciduous dentitions. It is structured into chapters according to the nature of the injury and whether injury has occurred in the primary or permanent dentition. At the end of each chapter there is a self-assessment questionnaire as well as references to relevant literature. Extensive use of pictures and video provides a comprehensive overview of the subject. PMID- 9855800 TI - Effect of systemic tetracycline and amoxicillin on inflammatory root resorption of replanted dogs' teeth. AB - The use of systemic penicillin after an avulsion injury has been recommended as a way to decrease the occurrence of resorption complications. Our previous study has shown systemic tetracycline to possess anti-resorptive properties independent of potential anti-bacterial actions. The purpose of this study was to compare histologically the effectiveness of tetracycline and amoxicillin in limiting inflammatory root resorption (IR) secondary to pulpal infection, in replanted dogs' teeth. Thirty root canals in five dogs were infected with plaque. The roots were extracted, the lingual mid-roots were shaved with a fissure bur ensuring cemental damage and immediately replanted. Dogs were given either tetracycline hydrochloride or amoxicillin, administered orally, on the day of extraction and replantation and for the following 6 days. The control group was teeth in these animals treated in the same manner but where no antibiotics were given. After approximately 6 months, the dogs were sacrificed and the teeth prepared for histologic evaluation. The shaved root surface was identified in cross-sections and eight equidistant points along the cut root surface were evaluated for healing (H) or IR in eight sections per tooth. For the tetracycline group, H averaged 67.22% and IR 32.78%, which was not statistically different from the amoxicillin group with H averaging 56.88% and IR 43.12%. The tetracycline group was superior to the control group with H = 27.86% and IR = 72.14%. Since tetracycline has been shown to have anti-resorptive properties in addition to the antibacterial properties demonstrated here, it could be considered as an alternative to amoxicillin after avulsion injuries. PMID- 9855801 TI - Effect of soaking in Hank's balanced salt solution or milk on PDL cell viability of dry stored human teeth. AB - This study was designed to evaluate the effect of soaking in either Hank's balanced salt solution (HBSS) or milk on periodontal ligament (PDL) cell viability in avulsed teeth. Dry storage times of 30, 60, and 90 min were evaluated. PDL cell viability was determined after removal of the cells from the root surfaces of extracted teeth using a modification of the procedure described by Nakashima (Arch Oral Biol 1991;36:655-63). After trypsinization and subsequent treatment in collagenase, the cells were stained with trypan blue, and viable and non-viable cells were counted using a hemocytometer and converted to percentages for statistical comparison. The results of this study demonstrated no significant difference in the number of viable cells with or without soaking in HBSS or milk at any of the dry storage times. In addition, there was no significant difference in PDL cell viability between the 30-and the 60-min dry periods. Although the soaking procedure had no obvious negative consequence, no significant improvement in PDL cell viability by the addition of this step was demonstrated under the conditions of this study. PMID- 9855802 TI - Susceptibility of Nd:YAG laser-irradiated root surfaces in replanted teeth to external inflammatory resorption. AB - Nd:YAG laser-induced modification of the root surface may inhibit development of external inflammatory resorption in replanted teeth. This study tested this hypothesis in vivo. The pulp chambers of six mandibular premolars in each of two dogs were accessed, inoculated with plaque, and sealed (Groups 1, 2). Two additional premolars in each dog were endodontically treated without inoculation (Groups 3, 4). After 2 weeks, teeth were hemisected and extracted. Each root had a 2 x 3 mm surface area denuded of cementum on the buccal and lingual surface. In Groups 1 (n = 12 roots) and 3 (n = 4), the denuded surfaces were wiped with 15% EDTA, coated with black ink, and irradiated with Nd:YAG laser (0.75 W, 15 pps, 300 microns tip, 20 s). In Groups 2 (n = 12) and 4 (n = 4), the surfaces were wiped with 15% EDTA, and rinsed with sterile saline for 20 s. Roots were replanted within 5 min. The dogs were perfusion-euthanised 10 weeks after replantation. Block specimens were removed, decalcified, embedded and horizontally sectioned (6 microns) at 180-microns intervals, resulting in 10 to 14 cross-sections of each root. From these, the middle five consecutive sections were stained with hematoxylin and eosin, and observed by light microscopy for occurrence of surface, inflammatory and replacement resorption on the denuded surfaces. No obvious differences were noted between the laser-irradiated and non irradiated surfaces. Inflammatory resorption was frequent in Groups 1 and 2, and absent in Groups 3 and 4. Replacement resorption was minimal in Groups 1 and 2, and frequent in Groups 3 and 4. Differences between Groups 1 and 2, and between Groups 3 and 4 were not significant, whereas the differences between the two pairs of groups were statistically significant (chi-square and two-way ANOVA, P < 0.006). These results did not support the hypothesis, and questioned the clinical validity of the surface modification in Nd:YAG laser-irradiated dentin. Therefore, the clinical application of Nd:YAG laser to the root surfaces of replanted teeth is not warranted. PMID- 9855803 TI - Local dexamethasone improves periodontal healing of replanted dogs' teeth. AB - The purpose of the present investigation was to evaluate histologically the effect of dexamethasone on root resorption in stored and replanted dogs' teeth. Twenty-nine roots from three beagle dogs were endodontically treated to prevent subsequent inflammatory root resorption of pulpal origin. The teeth were extracted and randomly assigned to three groups for 48-h storage. Group 1: control group teeth (n = 13) were stored in vials containing ViaSpan only; Group 2: topical treatment teeth (n = 10) were stored in vials containing ViaSpan and dexamethasone (16 micrograms/ml); and Group 3: systemic treatment teeth (n = 6) were stored in vials containing ViaSpan without any additives. Dexamethasone was administered intramuscularly (0.5 mg/kg body weight) 2 days prior to, on the day of, and every other day after extraction and replantation for two administrations. After 12 weeks, the dogs were sacrificed, and the teeth histologically prepared and evaluated according to the analysis of Andreasen. Significant differences were found for complete healing (P = 0.0583) and inflammatory root resorption (P = 0.0568) but not for replacement resorption (P = 0.1952). In addition, comparing local to systemic administration of dexamethasone (Group 2 vs. Group 3), statistically significant differences were found for healing, 85% (Group 2) vs. 67% (Group 3) (P = 0.0125) and inflammatory root resorption, 13% (Group 2) vs. 28% (Group 3) (P = 0.0126). This study indicated that topical use of dexamethasone enhances healing and results in fewer resorption complications. PMID- 9855804 TI - Calculus-like deposit at apex of tooth with refractory apical periodontitis. AB - It is generally accepted that bacteria in or outside the root canal are the reason for apical periodontitis and endodontic failures. This case report presents a 60-year-old woman with a periapical lesion and a fistulous tract which did not respond to conventional root canal treatment. During periapical surgery, granulomatous tissue was removed and a calculus-like deposit was observed on the root surface. A radicular cyst was diagnosed. Nine months after this calculus like deposit had been removed and the cyst enucleated, complete recovery of the bone had occurred. It is suggested that the presence of the calculus-like deposit at the apex of the tooth or its effects may in part have delayed the healing of the periapical inflammation in spite of apparently adequate endodontic treatment. PMID- 9855805 TI - Root resorption and apical breakdown during orthodontic treatment of a maxillary lateral incisor with dens invaginatus. AB - A 13-year-old girl was referred for endodontic treatment of a maxillary right lateral incisor with root resorption and apical radiolucency after orthodontic treatment. Radiographically the tooth had an invagination canal (Oehlers' Type III) and responded positively to pulp testing. The invagination canal extending to the lateral resorption was opened. Vital bleeding tissue was found, and the canal was cleaned and obturated. The pulp in the main root canal remained vital. The radiolucent lesion gradually decreased during the 3-year follow-up. The bone resorption seen here, despite vital tissue in the invagination canal, was probably due to osteoclastic activity caused by a low grade chronic infection, influenced by an injury-induced inflammation in the periodontium during orthodontic treatment. PMID- 9855806 TI - External influences on dental schools and their curricula. The influence of dental research on qualifications of clinical faculty staff. PMID- 9855807 TI - The Malmo model: a problem-based learning curriculum in undergraduate dental education. AB - A new undergraduate dental curriculum was introduced in Malmo in 1990 which is based on problem-based learning (PBL). The principles of PBL are strongly influenced by evidence from cognitive psychology and they form the educational strategy throughout the whole curriculum. Two further essential principles underpin the curriculum: a holistic attitude to patient care, and the promotion of oral health. Basic sciences and clinical dentistry are integrated within a structure based on oral conditions prevalent in the community. Students are encouraged to build their new knowledge, understanding and skills into the context of what they have already learned. This approach is facilitated by the presentation of conceptual models, one of which, The Oral Ecosystem is described in detail. The educational programme also gives students opportunities to learn in the clinical context from an early stage, and we endeavour to promote a scientific attitude from the very beginning of the programme. This paper describes the curriculum at three organisational levels (whole curriculum, single course, individual week). PMID- 9855808 TI - The basic science teaching experience in the Nordic countries. AB - The article concerns the dental education at 10 different faculties in Denmark, Iceland, Norway, Sweden, and Finland. The information is based upon a questionnaire to the faculties concerning the definition, the structure and succession of the basic science subjects/courses/themes/topics in the dental curriculum, the time span of the basic science teaching, and the methods of teaching. Furthermore, there is information about integration among the basic science subjects and the coordination/integration between the basic science teaching and the clinical teaching. Finally, the teachers' educational background and the cowork with the medical education is elucidated. The main findings are: the basic science teaching is structured in different ways in the Nordic countries. In Reykjavik, Bergen, and Aarhus this teaching is subject specific, while in Copenhagen it is mainly subject specific, as it has few integrated courses. In Gothenburg, Stockholm, and Umea, the basic science teaching consists of a mixture of integrated courses and subject specific teaching. In Helsinki, Oslo and Malmo the basic sciences are theme and topic based, and for the two last mentioned institutions, integrated with the clinic, except at the faculty in Malmo. Here the basic science teaching is totally integrated in the clinical teaching throughout the study. Usually, the basic science teaching is placed mainly in the first part of the curriculum; however, in Umea, Copenhagen, and Oslo, some integration takes place. PMID- 9855809 TI - Teaching dental undergraduates behaviour change skills. AB - This paper describes an active learning-based education tool which enables dental students to learn preventive techniques relevant to patient dental health behaviour. 2 studies were conducted involving 33, 2nd year (study 1) and 9, 3rd year (study 2) undergraduate dental students. In study 1, snacking behaviour and its antecedents were analysed from detailed 3-day diet diaries completed by the students. Study 2 entailed the students changing one aspect of their sugar/diet behaviour using self-management techniques. It is concluded that dental students can successfully (a) identify antecedents to sugar snacking behaviours on several levels, i.e., cognitive, emotional and situational, (b) set goals and use behaviour change techniques to modify these behaviours, and (c) appreciate that this experience is relevant to similar preventive techniques that they will use in clinical practice. Training in the application of these skills to their own maladaptive behaviours provides a strong educational tool based on psycho educational theories. PMID- 9855810 TI - Evaluation of a teaching aid for dental students. AB - A new teaching aid to assist undergraduate dental students to learn charting methods was developed. The aim of this study was to evaluate the effectiveness of the aid. Baseline charting tests were completed by 58 inexperienced clinical students at the start of a weekly half-day attachment at 3 community dental service (CDS) teaching clinics. These students were randomly assigned, in their tutor groups, to act as either test or control groups. The test group was actively taught charting with the new aid over their first 5-week attachment and control students were taught in the traditional way. Final charting tests were completed at the end of the attachments by all students and their results compared with baseline test results. The test group improved their mean scores from 20 out of a possible 50 marks at baseline to 36 at final test, a mean increase of 15 marks and an improvement of 71%. The control group scored a mean of 18 marks at baseline and improved this to 25, an increase of 7 marks, an improvement of 39%. It is concluded that the new teaching aid is more effective in assisting the students to learn charting than the traditional method. PMID- 9855811 TI - Providing care in an infectious disease clinic. Why students volunteer. AB - This qualitative study explores the motivations of dental students who volunteered as care providers for patients identified as carriers of infectious diseases. The study involved 12 students, seven 4th year and five 3rd year students who worked 1-2.5 days per week in an infectious disease clinic in a dental school in the northeastern United States. 4 faculty members who also volunteered their time in the clinic supervised the students. Personal interviews were conducted with each student to determine their reasons for volunteering and to explore the perceived significance of their experiences in the clinic. Internal validity was established through group interviews wherein responses during the personal interviews were discussed with the students. All 12 students were interested in Oral Medicine and all 4th year students had completed a course on HIV-AIDS for their selective requirement. Students expressed willingness to care for patients who are medically compromised in anticipation of an increase of this type of patients in their future practice. Previous exposure to a family member or friend or patients (non-dental) who were terminally ill or to patients with HIV was another common reason. All 12 students preferred to work in the infectious disease clinic more than in other school clinics. They felt that there was more faculty-student interaction and a more stimulating learning atmosphere that the faculty created. The students felt that the patients treated in the clinic were more appreciative. Students also claim they experienced more team spirit among their fellow students and staff and that the clinic atmosphere was more relaxed and less competitive. Students got the most satisfaction from helping patients who were perceived to have had problems in accessing care prior to their visit to the clinic. Results of the study show that the students volunteered to treat patients who are known to have infectious diseases such as HIV/AIDS because they find personal and academic satisfaction from working in a clinic which provides care to individuals with specific medical needs. PMID- 9855812 TI - 1993 AAFO clinician of the year. PMID- 9855813 TI - Postural assessment in dental orthopedics. PMID- 9855814 TI - Mixed dentition arch analysis. PMID- 9855815 TI - Past, present and future of functional orthodontics. PMID- 9855816 TI - Case 5. Refractory toothache. PMID- 9855817 TI - Microleakage of endodontically treated teeth restored with posts. AB - A fluid filtration system using 15 psi of pressure on the penetrating fluid was used to quantify the amount of microleakage of a stainless-steel post and a carbon-fiber post system, each placed with various cements. Statistical analysis showed that there was a significant difference in microleakage between the cements (p < 0.001). Zinc phosphate cement showed the most microleakage, whereas C & B Metabond cement showed the least. There was no significant difference in microleakage between the stainless-steel and carbon-fiber posts. The results of this study showed that both posts, when cemented with dentin-bonding resin cements (C & B Metabond and Panavia-21), exhibited less microleakage than when cemented with non-dentin-bonding cements (glass ionomer and zinc phosphate). PMID- 9855818 TI - Thrombin-induced DNA synthesis of cultured human dental pulp cells is dependent on its proteolytic activity and modulated by prostaglandin E2. AB - To clarify the roles of alpha-thrombin and prostaglandin E2 (PGE2) in the healing and inflammatory processes of dental pulp, their effects on the DNA synthesis of human pulp cells were investigated by measurement of [3H]thymidine incorporation. At a concentration range of 1 to 25 units/ml, alpha-thrombin stimulated DNA synthesis of the pulp cells by 1.5 to 2.6-fold. On the contrary, PGE2 (> 0.05 microgram/ml) suppressed DNA synthesis by 24 to 39%. Using reverse transcriptase polymerase chain reaction, thrombin receptor mRNA expression was identified in the pulp cells. Furthermore, alpha-thrombin-induced DNA synthesis could be inhibited by antithrombin III (2 units/ml) with heparin (2 units/ml) or D-Phe-Pro ArgCH2Cl (50 micrograms/ml). PGE2 (0.1 to 0.5 microgram/ml) also inhibited the thrombin-induced DNA synthesis by 39 to 64%. These results imply that pulp cells express the thrombin receptor that is activated by the serine protease activity of thrombin. Interactions of thrombin and PGE2 are important in modulating the inflammatory and healing processes of the pulp. PMID- 9855819 TI - Photoelastic comparison of stress induced by using stainless-steel versus nickel titanium spreaders in vitro. AB - The vertical force applied to an endodontic spreader generates stress along the canal walls. Recognizing the potential for this stress may reduce the incidence of vertical root fractures. A photoelastic acrylic model was fabricated to exhibit the stress produced during obturation of curved canals using the lateral condensation technique. Twenty standardized models simulating curved canals (32 degrees) were formed within PL-2 photoelastic acrylic resin blocks. The canals were fitted with a gutta-percha point, and either a stainless-steel or nickel titanium finger spreader was inserted. An Instron 4502 universal testing machine applied a vertical force of 20 Newtons to the spreader. Quarter wave and polarizing filters were used with backlighting to generate the fringe patterns in the models. Photographs of the resulting stress lines showed that the stainless steel spreaders created three areas of concentrated stress. The nickel-titanium spreaders induced stress patterns spread out along the surface of the canals, thus reducing the concentration of stress and the potential for vertical root fracture. PMID- 9855820 TI - Longitudinal study of the microleakage of two root-end filling materials using a fluid conductive system. AB - A comparison of the microleakage of mineral trioxide aggregate (MTA) and a high copper admix amalgam (Valiant PhD) in root-end preparations was made using a fluid conductive device. Thirty-three bilaterally matched pairs of extracted, single-rooted teeth were prepared and obturated using lateral compaction of gutta percha. The root-ends were resected and 3-mm-deep class I cavity preparations were made. The root-end preparations were filled with either amalgam or MTA. Guttapercha coronal to the root-end fillings was removed leaving only the amalgam or MTA present as a barrier to fluid movement. In the fluid-conductive device, the root canals were filled with a phosphate-buffered saline solution at a pressure of 10 psi. The flow of fluid was measured and compared at 1, 2, 3, 4, 8, 12, 16, 20, and 24 wk. The results showed amalgam to have significantly higher microleakage after 4 wk and higher variability compared with the MTA group. PMID- 9855821 TI - Effects of resorbable membrane placement and human osteogenic protein-1 on hard tissue healing after periradicular surgery in cats. AB - Periradicular surgeries were performed on the maxillary cuspid teeth of twelve cats. Before reapproximation of the surgical flaps, eight of the osteotomies were covered with a resorbable membrane and eight were filled with human osteogenic protein-1 (hOP-1) on a collagen carrier. The remaining eight sites received no further treatment and served as controls. The animals were euthanized after 12 wk, and the specimens were examined histomorphometrically for the presence or absence of osseous regeneration, inflammation, and cementum formation on the root ends. The results showed that the sites treated with the membrane exhibited significantly more inflammation adjacent to the resected root ends (p < 0.05), and that the use of the membrane had no statistically significant effect on osseous healing or new cementum formation. The use of hOP-1 was associated with a significant decrease in the thickness of new cementum formed on the resected root ends (p < 0.05), but had no statistically significant effect on osseous healing or degree of inflammation. Based on these results, it seems that neither the use of hOP-1 nor resorbable membranes have a positive effect on periradicular tissue healing in endodontic surgery. PMID- 9855822 TI - Ultrasonic root-end preparation: influence of cutting angle on the apical seal. AB - Apical resection is fundamental in obtaining a correct apical seal. The purpose of this study was to evaluate how the apical root resection angle and the cavity made by ultrasonic retrotips may influence the apical seal. A total of 48 extracted teeth were endodontically treated and sealed by guttapercha vertical compaction: 24 were resected with a 45 degrees angle and 24 with a 90 degrees one. An ultrasound source (P.M. 400 EMS) and a CT5 Scaler were used to make the retrograde cavity that was filled afterwards with EBA-zinc oxide-eugenol alumina added cement. Apical leakage was determined using fuchsin and assessed after the roots were sectioned longitudinally. Linear dye penetration in dentin and at the interface between dentin and cement was measured with a stereomicroscope (x 12 magnification), and the results were statistically analyzed. Results showed that there was less infiltration both in dentin and in the space between the filling and the dentinal wall in the group with the 90 degrees angle, but this difference was statistically significant (p < 0.0001) only for the dentin. None of the samples showed leakage greater than the depth of the preparation. An apical cavity of 3 mm or more along the vertical axis can produce a safe and effective seal. PMID- 9855823 TI - Mechanical properties of nickel-titanium endodontic instruments and the effect of sodium hypochlorite treatment. AB - The purpose of this study was to assess the mechanical properties--torsional moment, maximum angular deflection, maximum bending moment, and permanent angular deflection--of four brands of nickel-titanium (NiTi) endodontic file, and compare them with a conventional stainless-steel instrument, both in the presence and absence of sodium hypochlorite (NaOCl). NiTi instruments from four manufacturers were randomly selected and subjected to NaOCl treatment for 12 or 48 h, or not at all. The mechanical properties under test were then measured automatically by a digital torque memocouple. Torsional moment and maximum angular deflection indicate the resistance to torsional fracture of an instrument, maximum bending moment the stiffness of the instrument, and permanent angular deflection the strength of the base alloy. All instruments evaluated complied with or exceeded ADA/ANSI Specification No. 28, with the sole exception of the Maillefer ISO size 40 for torsional moment. JS Dental and McSpadden NiTi files were the most resistant to torsional fracture, but all NiTi files were inferior when compared with stainless-steel files from a previous study. However, NiTi files were superior in flexibility, and Maillefer and Brasseler instruments were the best of the instruments tested. NiTi files also had negligible permanent deformation angles. Furthermore, for all properties tested, NaOCl had no statistically significant effect. PMID- 9855824 TI - Cutting efficiency of nickel-titanium endodontic instruments and the effect of sodium hypochlorite treatment. AB - The purpose of this study was to assess the cutting efficiency of nickel-titanium (NiTi) files in the presence and absence of sodium hypochlorite (NaOCl) treatment, and compare them to a conventional stainless-steel K-type file. NiTi files from four manufacturers were randomly selected and exposed to NaOCl for 12 or 48 h, or not at all. Their cutting efficiencies were then assessed, using a new methodology: "mass of Plexiglass cut per energy used over 50 linear cutting motions." In the absence of NaOCl, Brasseler (318 micrograms/J) and Maillefer (280 micrograms/J) NiTi files were most efficient, followed by JS Dental (71.4 micrograms/J) and McSpadden (40 micrograms/J). These differences were significant (p < 0.0001), except for those between the latter two brands. Moreover, NaOCl treatment did not alter the cutting efficiency of any brand of instruments significantly. When compared with conventional stainless steel files, all NiTi files tested were less efficient. PMID- 9855825 TI - A photoelastimetric analysis of stress induced by root-end resection. AB - The aim of this study was to place into perspective some factors that played an important role in the healing of periapical lesions after surgical endodontic procedures. Root-ends were sectioned at different angulations on models and analyzed by photoelastimetry. Results showed that a section of the root-end through a plane perpendicular to the long axis of the tooth offers a better distribution of the stresses exerted on the apical region that the use of an inclined plane. The biological implications of these results have led the authors to propose a plan of resection that is perpendicular to the long axis of the tooth instead of the inclined plane commonly used. PMID- 9855826 TI - Reparative dentin formation in rat molars after direct pulp capping with growth factors. AB - Growth factors involved in normal wound healing may promote tissue repair when applied as a direct pulp capping medication. A minimal pulp exposure was made in rat molars, a pulp capping medication was placed, and the cavity was sealed. Epidermal growth factor, basic fibroblast growth factor, insulin-like growth factor II, platelet-derived growth factor-BB, and transforming growth factor-beta 1 (TGF-beta 1)--each absorbed onto a sterile collagen membrane (BioMend; Calcitek, Carlsbad, CA)--were used separately as pulpal medicaments. Dycal, unimpregnated collagen membrane, and no medication were used as controls. Eight samples from each treatment regimen were collected 2 and 3 weeks after surgery. Pulpal soft and hard tissue responses were graded. Data were analyzed by one-way ANOVA and Tukey-Kramer tests. No significant differences were detected after 2 wk. Pulp treated with TGF-beta 1 showed significantly improved soft and hard tissue healing at week 3, compared with the procedure control. We conclude that TGF-beta 1 as a pulp-capping medication enhances reparative dentin formation in rat molars. PMID- 9855827 TI - Nickel-titanium versus stainless-steel finger spreaders in curved canals. AB - Placement of endodontic spreaders to within 1 to 2 mm of the root canal working length has been advocated for optimum gutta-percha obturation. Due to their stiffness, stainless-steel (SS) spreaders will often fail to achieve this position in curved canals. Newly marketed nickel-titanium (NiTi) spreaders may offer an advantage in this regard due to the increased flexibility of these instruments. Seventy mesial and distal mandibular molar canals were instrumented to a #35 master apical file size. Measurements of the canal curvature were made from preoperative radiographs. Canal curvature angles ranged from 0 to 51 degrees. The teeth were placed in a radiographic mount to ensure a constant focal length, and postoperative radiographs were exposed with comparably sized SS and NiTi spreaders inserted into each prepared canal. Measurements of spreader tip to apex distance were made with a micrometer to the nearest 0.02 mm on the radiographs. Using a t test for paired samples, it was shown that NiTi spreaders penetrated to a significantly greater depth than SS spreaders in curved canals (p < 0.001). PMID- 9855828 TI - Changing number and distribution of endodontists: a continuing imbalance--1987 1995. AB - A review of the 1995 and past American Dental Association studies on the distribution of dentists provides specific information on the evolving numbers of endodontists at the national, regional, and state levels. Despite an increase in numbers of endodontists, major differences continue in practitioner-to-population ratios in different geographic areas. PMID- 9855829 TI - Rubber latex allergy and the endodontic patient. AB - Increasingly, patients with rubber latex allergy are being seen in dental offices. Health care workers, persons allergic to certain foods and those with certain medical histories are potentially latex-sensitive. This article presents the case of a patient with a history of severe rubber latex allergy and the associated management of an endodontically involved tooth. The possibility of the latex allergic patient being sensitive to gutta-percha obturation material is also raised. PMID- 9855830 TI - Bacterial reduction with nickel-titanium rotary instrumentation. AB - The purpose of this study was to compare intracanal bacterial reduction on teeth instrumented with 0.04 tapered nickel-titanium (NiTi) rotary instrumentation to bacterial reduction when using a stainless-steel K-file step-back technique using sterile saline irrigation. Forty-eight patients with apical periodontitis were randomly assigned treatment type. The canals were sampled before, during, and after instrumentation. The samples were incubated anaerobically for 7 days at 37 degrees C, colony-forming unit numbers calculated, and a log transformation performed to normalize the counts. Teeth exhibiting apical periodontitis were uniformly infected, whereas vital control teeth were not. A similar and uniform reduction occurred with progressive filing, regardless of technique (p < 0.0001). There was no detectable difference in colony-forming unit count after NiTi rotary or stainless-steel hand instrumentation (p = 0.42). Neither technique could predictably render canals free of bacteria. The results of this study indicate NiTi rotary and stainless-steel hand K-file step-back instrumentation techniques were not significantly different in their ability to reduce intracanal bacteria. PMID- 9855831 TI - Evaluation of setting properties and retention characteristics of mineral trioxide aggregate when used as a furcation perforation repair material. AB - Furcation perforations were created in 32 extracted maxillary and mandibular molars. The perforations were prepared in the center of the pulp chamber floor parallel to the long axis of each tooth and a saline-moistened Gelfoam matrix was placed below the perforation to simulate a clinical condition. The teeth were randomly divided into four groups, and the perforations were all repaired with mineral trioxide aggregate (MTA) and then covered with either a wet or dry cotton pellet for 24 or 72 h. Instron testing was used to measure the force required to displace the material from the perforation. The force measurements showed that MTA resisted displacement at 72 h to a significantly greater level than at 24 h (p < 0.05). When slight displacement occurred at 24 h the material demonstrated the ability to re-establish resistance to dislodgement from the dentin wall. The presence of some moisture in the perforation during placement was advantageous in aiding adaptation of MTA to the walls of the perforation, but there was no significant difference in MTA retention when a wet or dry cotton pellet was placed in the pulp chamber during the setting time (p > 0.05). PMID- 9855832 TI - HIV and the family. PMID- 9855833 TI - AIDS cases soar in past year. AB - Around 1.5 million people developed AIDS between mid-1993 and mid-1994--three times as many as in the previous 12 months, according to estimates released by GPA on 1 July. About 200,000 of the new cases are thought to be in south and southeast Asia, more than eight times as many as occurred in that region during the previous year. PMID- 9855834 TI - Ten points for world AIDS day 1994 AIDS and the family. PMID- 9855835 TI - Sexual behaviour research and HIV/AIDS. AB - To design effective ways of reducing HIV transmission through sexual intercourse, which accounts for about 90% of all infections among adults and adolescents, we need to know more about how sex occurs and is understood in different cultures. Dr Peter Aggleton, who leaves GPA this summer after two years as head of its social and behavioral studies and support unit (SSB), explains the Programme's work in this area. PMID- 9855836 TI - Labial gland biopsy--a simple diagnostic procedure for Sjogren's syndrome. AB - Sjogren's syndrome is a collagen vascular disorder with multisystem. Its presentation may be similar to other diseases from which it can be differentiated by certain investigations--one of the most important being the labial salivary gland biopsy. A case report of Sjogren's syndrome is presented and the technique of the labial salivary gland biopsy is described. PMID- 9855837 TI - Condylocoronal angle of the human mandible--its radiological implications. AB - 26 dry specimens of mandible were utilised for making antropometric & condylo coronal angle measurements. 26 living subjects were radiographed in a standardized SMV an CCA values were established. Average value of Antero posterior dimension was obtained to be 8.5 mm and latero medial dimension obtained to be 17 mm. Left and right condylo coronal angle (CCA) from dry specimens was assessed to be 18 & 16 respectively. From the living subjects CCA was established as 17 degrees in Indian population. The following radiological implications are evident from this study. (1) A tomographic cut of the TMJ if done at the depth of 8 to 9 mm i.e. half of 17 mm (lateromedial dimension average) it would pass through the centre of the condyle and may contribute to the diagnostic importance. (2) The CCA of 17 degrees obtained for Indian population may be utilized by clinicans to obtain clearer transpharyngeal and transcranial views of the TMJ by adjusting the radiographic cone to 17 degrees tilt in horizontal angulation keeping the standard values for vertical angulation as--10 degrees for transpharyngeal and +25 degrees for transcranial views. PMID- 9855838 TI - A survey of 4478 biopsy specimens of oral lesions. AB - A surgery of 4478 biopsy specimens in dental institute in India was carried out to compare and correlate results obtained from such studies carried out elsewhere. The study revealed that there are more than 150 common and rare oral diseases which are diagnosed and treated by dentists. 13 common diseases comprised majority (60.37%) of the biopsy specimens studied. Of these, tumors (25.53%) and Cysts (14.38%) constituted the majority of the bulk. The study also shows that most of the benign tumors were mesenchymal in origin (almost 5.5:1) while malignant tumors were epithelial (almost 20:1) in origin. Cystic lesions included the most common radicular cyst (7.12%) and the rare hydatid cyst (one case reported). Gingival hyperplasia mainly due to local aetiology, constituted the 3rd largest group. The purpose of this report is to provide some information about the type and frequency of oral lesions usually diagnosed, with the help of histopathological examination in India. PMID- 9855839 TI - The role of mandibular third molars in relation to lower anterior crowding--a pilot study. PMID- 9855841 TI - What can organized dentistry do for me? PMID- 9855840 TI - International President's message. PMID- 9855842 TI - Cardiovascular disease and periodontal disease. PMID- 9855844 TI - Insurance overpayments ... do you have to return? PMID- 9855843 TI - Unmasking patient attitudes: the case for patient immersion. PMID- 9855845 TI - What you can learn at your bakery to preserve fee-for-service dentistry. PMID- 9855846 TI - Mentorship Program. PMID- 9855847 TI - What happens if I'm sued. PMID- 9855848 TI - Problem-based learning: not a new paradigm. PMID- 9855850 TI - Clinical evaluation of a resin-modified glass-ionomer adhesive system. AB - One hundred cervical caries-free, nonundercut lesions were restored in 13 patients of mean age 60.5 years, using Silux Plus or Estio LC resin composite bonded with Fuji Bond LC. Patients were recalled at 6 months and 1 year. A 1-year cumulative retention rate of 98% was recorded, while marginal staining was absent. PMID- 9855849 TI - An in vitro study of the effect of restorative technique on marginal leakage in posterior composites. AB - The purpose of this in vitro study was to compare marginal leakage of class 2 light-cured composite resin restorations, with different insertion and polymerization techniques. Fifty sound premolars extracted for prosthetic purposes were used in the study. Conservative class 2 (MOD) preparations were made in all teeth, with one proximogingival margin on enamel and the other on cementum. All the preparations and restorations were performed by the same operator utilizing the same restorative materials. After restoration the teeth were thermocycled and their gingival margins exposed to a dye. Results showed that the kind of matrix system had no significant influence upon the degree of marginal leakage on enamel. On cementum, oblique and horizontal incremental insertion and polymerization techniques using the collimator cone exhibited the least leakage, while the clear matrix band/reflective wedge technique presented the greatest leakage. PMID- 9855851 TI - Dyes for caries detection influence sound dentin bond strength. AB - This study evaluated the influence of caries-detection dyes on the in vitro tensile bond strength of adhesive materials to sound dentin. Caries-free human molars were ground to expose superficial dentin. Two dyes (a commercial 0.5% basic fuchsin in propylene glycol and Cari-D-Tect) were applied to sound dentin and rinsed. Subsequently, the dentin was etched with phosphoric acid (35%) and rinsed, leaving a moist dentin surface. The adhesive (Prime & Bond 2.0) was applied in two layers and light cured. A composite (TPH Spectrum), a compomer (Dyract), and a hybrid ionomer (Advance) were used to prepare the bond-strength specimens with a 3-mm-in-diameter bonding area. Control groups were made without use of dyes. Six specimens were prepared for each group. After 24 hours in distilled water, tensile bond strength (MPa) was measured using a testing machine. Analysis of variance was used to evaluate the data. Without dyes, bond strengths of TPH Spectrum and Dyract with Prime and Bond 2.0 were similar and both values were significantly (P < 0.05) higher than that of Advance with Prime & Bond 2.0. Dyes for caries detection reduced the bond strength of TPH Spectrum and Dyract but not Advance when used with Prime and Bond 2.0. PMID- 9855852 TI - The heat sink and its relationship to reducing heat during pin-reduction procedures. AB - When self-threading pins are used in the restoration of teeth, the heat generated by cutting these pins to the correct length may traumatize the pulp. This study evaluated cotton forceps and a mosquito hemostat used as a heat sink and at different positions on the pin to determine any change in heat transferred to the tooth. Three hundred self-threading pins (Whaledent TMS System: 100 each of the Regular, Minim, and Minikin sizes) were inserted into freshly extracted teeth and sectioned. The temperature change at the apical end of the pin was measured using a thermistor probe. The results indicated that metal supporting instruments can be effective in reducing heat transferred to the apical aspect of the pin during pin reduction. PMID- 9855853 TI - Evaluation of custom occlusal matrix technique for posterior light-cured composites. AB - Posterior composite restorations are difficult to contour and polish due to their occlusal anatomy and opposing occlusion. Recently a new technique utilizing a clear custom occlusal matrix showed promise in reducing the chair time necessary to place posterior composite restorations. Sixty specially molded plastic mandibular right second molar teeth with a mesio-occlusal composite preparation were restored with a light-cured composite in a typodont, using either the custom occlusal matrix (experimental) or the standard technique (control) as taught at the University of Michigan School of Dentistry. The times necessary to complete the procedures were recorded. The teeth were evaluated in a blind manner by two evaluators using a modified Ryge criteria. The times to place and finish the control and experimental restorations were subjected to a t-test. The evaluations of surface smoothness, margin adaptation, occlusal anatomy, and axial form were compared using a Fisher exact test. The custom occlusal matrix significantly reduced placement and finishing time and improved surface smoothness of mesio occlusal posterior composite restorations placed in vitro. Due to the time needed to construct the custom occlusal matrix, there was no significant difference in total mean time of the two procedures. PMID- 9855854 TI - Short-term reaction of exposed monkey pulp beneath adhesive resins. AB - Monkey pulpal responses to four adhesive resin systems used as direct pulp capping agents were histopathologically evaluated at 3, 7, 14, 30, and 60 days after operation. No serious inflammatory reaction of the pulp, such as necrosis or abscess formation, was observed. Slight inflammatory cell infiltration was the main inflammatory reaction of the exposed pulp, and the exposed area became occluded with dentin bridging as the observation period increased. Healing of exposed dental pulp beneath adhesive resin capping slightly differed depending on the materials used. PMID- 9855856 TI - Loss of sound tooth structure when replacing amalgam restorations by adhesive inlays. AB - The replacement of amalgam restorations by adhesive inlays requires an adjustment to the cavity form. This often necessitates the removal of sound tooth substance. Undercuts may be blocked out by a base, but an extensive use of glass ionomer for this purpose is not recommended due to the weak mechanical properties of this material. The present study estimated the amount of sound tissue removed when a given amalgam preparation was reshaped for an adhesive inlay without the use of a base. An MOD amalgam preparation was created in an acrylic tooth. Twenty copies were distributed among 20 operators, who were requested to transform the preparation into an adhesive inlay preparation, removing as little material as possible. One control preparation with parallel walls was produced. All teeth were weighed before and after the alteration. The preparations' volumes were calculated: original amalgam preparation 0.130 ml, parallel preparation 0.136 ml. The minimum removal necessary was therefore 0.006 ml. The amount removed by the operators varied, with a mean volume of 0.0138 +/- 0.004 ml. Statistical analysis showed that for the given preparation, significantly more material was removed than necessary for undercut elimination. On average, more than twice the minimal volume of material was lost. This loss can be minimized by developing alternative techniques for undercut elimination. PMID- 9855855 TI - 6-year clinical evaluation of fired ceramic inlays. AB - The purpose of the present study was to evaluate the quality of fired ceramic inlay restorations consisting of a feldspathic porcelain system (G-Cera Cosmotech II) after 6 years in vivo. A total of 49 fired ceramic inlays (for 27 premolars and 22 molars: class 1 in 11 teeth, class 2 in 36 teeth, and onlays in two teeth) were placed in 29 patients, and all restorations were evaluated at the time of placement, at 6 months, and 1, 2, 4, and 6 years after placement using modified USPHS criteria. Replicas made of the restorations were observed using a scanning electron microscope (SEM) to assess the state of the marginal area and wear of the restoration. Favorable color match was noted, and no bulk fracture, tooth fracture, or missing restorations were detected. Longevity was observed in 92% of the fired ceramic inlay restorations at 6 years (Kaplan-Meier method); however, marginal fracture was detected in six restorations (13%) and marginal discoloration in 11 (23%). SEM evaluation disclosed marginal microfracture in 49% of the restorations, wear in 19%, and wear of resin cement along margins in 36% at 6 years. No significant difference was observed in the incidence of each of these three characteristics between molars and premolars. This longitudinal 6 year clinical observation suggested that fired ceramic inlay restorations made by the G-Cera Cosmotech II system are aesthetic, durable, and clinically acceptable. Further follow-up is needed for the restorations showing marginal degradation and fracture from a microscopic and macroscopic perspective. PMID- 9855857 TI - Foundations of genetics: genetic structure, function, and therapeutics. AB - Molecular genetics provides the basis for understanding patterns of health and disease in people and is part of the scientific foundation on which acute and critical care nurses should build their practice. The human genome, defined as all the genetic information in the cells of humans, provides the blueprint for protein production and cellular function in the body. Alterations in protein production may result in illness or organ malfunction that has a genetic derivation. One therapeutic strategy that holds promise to manage genetic diseases is gene therapy. Gene therapy, or human gene transfer, occurs when scientists or physicians modify the genetic material in cells for therapeutic purposes. Genetic structure, function, and therapeutic reflect the science of the present and future and have profound practice implications for acute and critical care nurses. PMID- 9855858 TI - Genetic family history assessment. AB - The purpose of this article is to familiarize nurses with why, how, when, and where a genetic family history assessment should be used in clinical practice. Pedigrees are diagrams that display the relationship among family members by using a combination of symbols and lines. They are used to record concisely a complete family history to identify the risk of transmitting inherited condition, to identify people at risk for development of adult-onset conditions, to aid in clinical diagnosis, and to serve as a reference for social and biologic relationships. A detailed explanation of the standardized pedigree nomenclature that was recommended by the Pedigree Standardization Task Force in 1995 is included. Step-by-step guidelines on taking a genetic family history and drawing pedigrees are provided. A case study is also included to illustrate pedigree construction. PMID- 9855859 TI - Molecular genetics: from bench to bedside. AB - Diligent work performed at the laboratory bench during the 20th century has resulted in advances in the health care industry and benefits for the patients it serves. Time-saving laboratory techniques such as DNA isolation and polymerase chain reaction (PCR) have helped molecular biologists and geneticists learn more about genes and their function. Information resulting from genetic research is currently used by medical researchers to develop genetic testing, genetic engineering, and gene therapy procedures that will benefit patients with genetic needs. This article provides basic information regarding several of these procedures, including DNA isolation, PCR, gel electrophoresis, and restriction enzyme techniques. In addition, the article explores the experiences of a clinical nurse, who by learning genetic laboratory techniques, developed an appreciation of the nursing implications related to genetic laboratory procedures. PMID- 9855860 TI - Optimizing drug therapy based on genetic differences: implications for the clinical setting. AB - Differences in drug responses due to gene alterations are rapidly being identified. Gene alterations may inhibit the function of an enzyme so that an active drug accumulates, causing adverse reactions with normal doses. Alternatively, gene alterations may accelerate enzymatic function so that an active drug is rapidly eliminated, causing subtherapeutic responses to normal doses. Mutations and polymorphisms have been identified that affect a person's response to many currently prescribed medications including cardiovascular, anti infective, chemotherapeutic, psychiatric, and analgesic drugs. The potential exists for drug therapy to be optimized by selecting medication and doses based on a person's genotype rather than by trial and error. In the near future, advanced practice nurses in the acute care setting may be expected to order, provide patient education about, and explain results of genetic tests before initiating a specific drug therapy. Advanced practice nurses must be knowledgeable about what genetic tests are analyzing and their benefits, limitations, and risks. PMID- 9855861 TI - The impact of Alzheimer disease genetics on expert and advanced gerontological nursing practice. AB - Alzheimer disease (AD), a progressive neurodegenerative disorder, is the most common cause of dementia in the United States, affecting as many as 4 million people. Extensive research is under way to identify environmental and genetic risk factors for this complex disease. Currently, four genes are associated with an increased risk for AD: the amyloid precursor protein gene on chromosome 21, the Presenilin I gene on chromosome 14, the Presenilin II gene on chromosome 1, and the apolipoprotein E gene on chromosome 19. Expert and advanced practice gerontological nurses are faced with new challenges as a result of these gene discoveries. Gerontological nurses should assess for relevant environmental and genetic risk factors; obtain comprehensive family health histories recorded as pedigrees; integrate genetic information into diagnosis, intervention, and evaluation strategies; initiate and coordinate referrals to genetic specialists; and provide ongoing emotional and decision-making support for patients and families experiencing AD. PMID- 9855862 TI - Genetics and the central nervous system: apolipoprotein E and brain injury. AB - Apolipoprotein E (apoE), a protein produced by glial cells, is responsible for maintenance of the structural integrity of the microtubules within the axon of the neuron. The gene associated with apolipoprotein E, APOE, influences the construction and regeneration of the microtubules in an APOE allele-specific manner: APOE 2/2 may be neuroprotective, whereas APOE 4/4 may be neurodestructive. Thus, APOE appears to be one genetic factor that modifies the brain's response to insult, and therefore may modify the severity of neuropsychologic deficits. This article presents an overview of the genetic relation between APOE and neuropsychological function in Alzheimer disease and proposes a relation between APOE and recovery after head injury. PMID- 9855863 TI - Genetic risk factors in the development of heart disease: familial hypercholesterolemia and hyperhomocysteinemia. AB - Cardiovascular disease continues to be the leading cause of death in the United States. Several risk factors for heart disease are genetically based. Two of these risk factors are familial hypercholesterolemia and hyperhomocysteinemia. Effective and relatively inexpensive treatments have been established for these risk factors. This article reviews the genetic basis for familial hypercholesterolemia and hyperhomocysteinemia, presents strategies for diagnosis and treatment, and discusses implications for nursing practice. PMID- 9855864 TI - The genetic basis of diabetes mellitus. AB - Familial risk, pathogenesis, clinical onset, and treatment of diabetes mellitus vary according to etiology. Although Type 2 diabetes has a higher familial risk, more is known about the genetics of Type 1 diabetes. Genes contributing 60% to 65% of susceptibility to Type 1 diabetes mellitus are known. Type 1 diabetes is associated with susceptibility genes in the HLA region on chromosome 6p21 and the insulin gene on chromosome 11p15, and at least eight additional susceptibility genes are under investigation. Islet cytoplasmic antibodies provide humoral evidence of Type 1 diabetes risk. Only 10% of the genes contributing susceptibility to Type 2 diabetes mellitus are known, and they are primarily associated with uncommon subtypes of the disorder. The insulin receptor gene on chromosome 19p13 and at least five glucose transporter genes contribute to Type 2 diabetes susceptibility, and further associations may emerge from study of the glycogen synthase gene, the glucokinase gene, the MODY genes, and the leptin gene. Diabetes comorbidities may result from genetic and environmental susceptibilities independently or in combination. PMID- 9855866 TI - Marfan's syndrome in pregnancy: implications for advanced practice nurses. AB - Marfan's syndrome is a genetic disorder that often affects the cardiovascular, pulmonary, ocular, and musculoskeletal systems. When a woman with Marfan's syndrome becomes pregnant, the hormonal stresses and changes to the cardiovascular system can put the mother at risk for serious complications. Advanced practice nurses need to understand the maternal changes associated with Marfan's syndrome as well as the genetic factors involved in order to provide holistic care. PMID- 9855865 TI - Genetic predisposition to breast and ovarian cancer: a case study. AB - Information concerning genetic predisposition and the discovery of genes associated with certain cancer risks is changing rapidly. Nurses must keep abreast of these changes so that they can better understand the choices patients have and the consequences of those choices. This article discusses the issues pertinent to women with a genetic predisposition to breast or ovarian cancer, or both. Discoveries in the Human Genome Project have already begun to change traditional perspectives on screening, diagnosing, preventing, and treating cancer. These genetic discoveries hold both promise and concern for health care professionals. The promise lies in the precise identification of genetic predisposition to common diseases and the potential to prevent or reduce morbidity and mortality rates. The concern lies in issues of confidentiality and discrimination: Predicting predisposition to incurable illnesses may have substantial negative impact on the person's quality of life and psychosocial integrity. PMID- 9855867 TI - Linking critical care nursing and genetics with research funding opportunities. AB - The purpose of this article is threefold: to reveal how a disease that afflicts many critical care patients can be the impetus for forging into a research career; to illustrate a program of research undertaken by a nurse investigator to answer a critical care nursing question using genetic technology; and to identify exciting opportunities for research training, career development, and investigator-initiated research activities for the advanced practice critical care nurse at the National Institute of Nursing Research. The article concludes by identifying future linkages between nursing research and genetics. PMID- 9855868 TI - Genetics on the World Wide Web. AB - Since 1990, when the Human Genome Project was initiated, the amount of genetic information on the World Wide Web (WWW) has grown substantially. The WWW has become an important resource for current, accurate, and reliable genetic information for health care professionals and the general public. The purpose of this article is to provide a variety of genetics-related WWW sites that are useful for all levels of practitioners interested in genetics. In selecting sites to be included in this article, a number of evaluation tools were reviewed. The primary concern was that these sites be reputable and provide accurate, timely information. A table of the WWW sites is included for quick easy reference. PMID- 9855869 TI - National coalition for health professional education in genetics. AB - With the explosion of genetic information, there has been a recognized need for more genetic knowledge among health care professionals. The National Coalition for Health Professional Education in Genetics (NCHPEG) was established in 1996 to address this need. This article briefly outlines the mission of this Coalition and gives an overview of its relationship to acute and critical care nurses. PMID- 9855870 TI - The application of genetic knowledge: ethical and policy implications. AB - This article explores societal ethical concerns about the application of the rapidly developing knowledge in the field of genetics. The article calls for nurses to reflect on these ethical concerns not only in the care of patients but also in professional and societal discussion, so that sound ethical and societally acceptable public policy can be adopted. A brief summary introduces the concepts of ethics and policy. The text than offers discussion of genetic testing for adults and children, testing in relation to behavioral attributes, and genetics and reproduction. Hypothetical cases using fictional names are used to illustrate each section. Recommendations for nurse action are cited. Finally, the article poses questions that require public awareness and discussion. PMID- 9855872 TI - Glossary of genetic terms. PMID- 9855871 TI - Genetic research and ethical challenges: implications for nursing practice. AB - Careful consideration of the potential consequences of biomedical research involving human beings is an important advanced practice strategy. The presence of disease and the hope for a cure through genetic manipulation create a complex mix of ethical concerns. In this article, some of the actual and potential issues associated with genetic research and the implications for nursing practice are discussed. The principles of autonomy, beneficence, and justice provide a framework for the discussion. PMID- 9855873 TI - The Agency for Health Care Policy and Research guidelines: implications for home health care providers. AB - The Agency for Health Care Policy and Research (AHCPR) was established in 1989 under the U.S. Department of Health and Human Services as a means to facilitate the development and implementation of clinical practice guidelines that are based on current research and the consensus opinion of experts within a specific area of clinical practice. This article provides a summary of the recommendations in each guideline with implications for home health care nursing practice. PMID- 9855874 TI - Graduate program: advanced practice nurses in the home. AB - This article is a curriculum overview of a graduate program of the Advanced Practice Home Care Nursing (APHCN) Program, which began at the School of Nursing, University of California, San Francisco, in 1996. A historical perspective of home care and the need for advanced practice nurses in the home care industry is provided. Curricular components of the APHCN Program are outlined. The roles of the home care clinical nurse specialist, home care nurse practitioner, and home care administrator are presented. Finally, a discussion is presented in which home care nursing demonstrates the efficacy of the advanced practice role and the need for well designed outcomes-based research. PMID- 9855875 TI - Antepartum home care for high-risk pregnant women. AB - Home care for women experiencing complications of pregnancy is a rapidly growing specialty. A variety of models of antepartum home care have emerged. Provision of effective antepartum home care requires knowledge and clinical skills in perinatal and home health nursing and an understanding of the structure and function of the home health care system. Nursing care in the home encompasses case coordination, maternal and fetal assessment, maternal education regarding the woman's risk situation and self-monitoring requirements, and provision of support. Antepartum home care for conditions such as hypertension in pregnancy, preterm labor, and hyperemesis gravidarum is discussed. PMID- 9855876 TI - Managing congestive heart failure at home. AB - The hospital readmission rate and associated cost of treating patients with heart failure continues to escalate. Hospitals now regard home care as an alternative to institutional care for mthe long-term management of heart failure. Heart failure management programs are becoming increasing popular in home care as home health agencies strive to meet the demands of the Medicare managed-care environment. Practice standards such as those of the Agency for Health Care Policy and Research (AHCPR), form a basis by which home health agencies can develop, implement, and evaluate their program. Heart failure management continues to improve with advances made toward decreasing morbidity and mortality. The cardiac home care nurse assumes a pivotal role in the management of these complex patients. PMID- 9855877 TI - Role of the advanced practice nurse in continence care in the home. AB - Because incontinence is one of the most common problems encountered in the home care setting, home care agencies are beginning to recognize the importance of having a continence care program, and the need for advance practice nurses (APNs) to assume roles as continence care nurses (CCNs) is increasing. Through a comprehensive continence care program, incontinence is curable and easy to treat. This article describes the role of the APN as a CCN, and provides a description of the assessment and therapeutic management of the various types of urinary incontinence, focusing on the elderly patient in the home care setting. PMID- 9855878 TI - Evolution of transitional care settings: past, present, future. AB - Transitional care settings have evolved since the 1980s as Medicare prospective payment has affected acute hospital reimbursement and hospital length of stay. These settings include hospital-based subacute units, typically called transitional care units; subacute care units in freestanding nursing facilities; and long-term care hospitals. This article focuses on these transitional care settings: what they are, how they developed, what reimbursement issues affect these settings, and what is likely to happen next. In the future, advanced practice nurses may not only refer patients to transitional care settings, but also may find them to be sites for practice. PMID- 9855879 TI - A case study in interdisciplinary care of the critically ill. AB - The special care unit (SCU) of University Hospitals of Cleveland was established in 1989 specifically for case management of critically ill patients who require long-term habitation in an intensive care setting, usually because of ventilator dependency. This case study exemplifies the interdisciplinary involvement in caring for a patient with complications who was a long-term, critically ill resident of the unit. The discussion shows the importance of the use of the nurse case manager coordinator and an interdisciplinary approach to patient care for successful outcomes in the chronically critically ill population. PMID- 9855880 TI - Swallowing in patients with tracheotomies. AB - Swallowing and respiration are well-coordinated and interdependent functions. When one of these processes is impaired, the ramifications may be negative for the other. This article describes the mechanics of normal swallowing, the disorders of swallowing, the effects of tracheotomy and mechanical ventilation on swallowing, and the procedures used to assess and treat swallowing. Combining a basic understanding of these concepts with practical management can increase safe and efficient oral intake in patients with artificial airways. PMID- 9855881 TI - Nutrition support for patients in long-term acute care and subacute care facilities. AB - Long-term acute care and subacute care facilities (also transitional care facilities) have evolved from the need to decrease costs associated with acute care in the hospital. As the length of stay in many medical centers has been reduced, patients are admitted to transitional care facilities to continue recovery and rehabilitation. Rehabilitation and recovery can be enhanced with the provision of optimal medical nutrition therapy. Nutrition screening is essential in identifying patients who are at risk of malnutrition or are malnourished. Nutrition assessment verifies the risk or presence of malnutrition followed by the development, implementation, and monitoring of nutrition intervention. Nutrition screening and intervention promote recovery from illness, minimize morbidity and mortality, and enhance quality of life. The goals of nutrition support are to prevent starvation-associated malnutrition, preserve lean tissue mass, support metabolic functions, and improve clinical outcomes. Oral nutrition is the preferred method of nourishment; however, specialized nutrition support is considered for patients unable to meet their nutrient requirements adequately. Enteral nutrition support is recommended when providing nutrition support however, parenteral nutrition support is used when the gastrointestinal tract can not be safely used. With appropriate intention, administration, and monitoring, nutrition support can be safely administered. PMID- 9855882 TI - Nutritional requirements of an aging population with emphasis on subacute care patients. AB - Life expectancy in the United States has increased dramatically during the past century, creating a large population of people with potentially compromised nutritional status. Although requirements were previously believed to decline with aging, recent data suggest that requirements for protein and energy often do not decline and may actually increase during disease exacerbations. Optimal intake of vitamins and minerals is also under reevaluation, and significant segments of the population may have deficiencies based on limited intake or absorption. Social and psychological factors and difficulty chewing and swallowing may interfere with adequate intake. At the end of a hospital stay, many patients need continued skilled transitional care before discharge home. Many patients who have existing nutritional deficits, including weight loss, decreased serum proteins and pressure ulcers are admitted to subacute nursing facilities. Careful initial and periodic reassessment of nutritional status and aggressive nutritional management must be used to prepare patients for optimal independence after discharge. Improved clinical outcome can be achieved with prevention of malnutrition and timely correction of nutritional problems. PMID- 9855883 TI - Bridging the gap: medical directives for acute care nurse practitioners. AB - The following article describes the process by which a group of acute care nurse practitioners sought to address the legal challenges of working beyond the traditional scope of nursing practice. It was necessary to establish mechanisms for communicating a diagnosis, as well as for ordering diagnostic tests, treatments and procedures. Medical directives were viewed as an approach to address components of practice involving controlled acts not authorized to nursing. The process of developing medical directives began with a description of the components of a medical directive. Algorithms were then developed based on the College of Nurses of Ontario's decision tree (Purvis, 1995) for the performance of procedures. These algorithms were broad and applicable across all clinical programs. The final step, required each nurse practitioner/clinical nurse specialist in collaboration with physician colleagues, to develop individual appendices specific to each clinical program. Health care administrators may find the information provided of assistance in addressing legal concerns that arise when new opportunities for nursing involve movement beyond traditional boundaries. PMID- 9855885 TI - Perspective: the Nursing Education Council of British Columbia. PMID- 9855884 TI - Factors influencing job satisfaction on specialty nursing units. AB - In the midst of downsizing, restructuring, layoffs, hospital closures, mergers, and the beginning cycle of shortages in specialty units, nursing administrators must extend their understanding of the factors influencing job satisfaction and the implications these findings may have for nursing practice, in order to enhance the quality of worklife for nurses in a hospital setting and create competitive work environments. The Causal Model of Job Satisfaction for Nurses (Blegen & Mueller, 1987), including Leatt and Schneck's (1981) technology variable, was the conceptual framework used to look at the effect of the 14 variables (opportunity, routinization, autonomy, job communication, social integration, distributive justice, promotional opportunity, motivation, pay, workload, general training, kinship responsibility, unit size, technology) on job satisfaction. This study demonstrated a statistically significant positive correlation between autonomy, motivation and job satisfaction and a statistically significant negative correlation between routinization and job satisfaction. PMID- 9855886 TI - Partnerships: the foundation for future success. AB - Partnerships will most probably be considered the watchword of the nineties. Their meaning is extrapolated from interviews with individuals involved in creating partnerships of all kinds as well as reviewing selected literature. Criteria for success and common themes found in failure are presented. The relationship between collaboration and partnership is discussed as well as the impact on power bases. Because partnership is fundamentally viewed as a process entailing the development of relationships, two author's views of developmental sequences are compared and contrasted. PMID- 9855887 TI - Health care issues: managing services and people in home care: today's challenge. AB - This article is the second in a series of three focusing on key management processes in home care. The first article addressed the issue of quality management and the next article looks at financial and information management. This article examines the issues of service and human resource management with an emphasis on service management. It starts by outlining some of the pressures behind the unprecedented demand for home care services and looks at a few of the strategies home care agencies can utilize to manage service demand. It then moves to a discussion of human resources, specifically the case manager. Basic qualifications, knowledge, skills, and attributes are outlined and administrators are encouraged to facilitate the ongoing development of this costly and critical resource. PMID- 9855888 TI - Witmer establishes provincial task force to review nursing services. PMID- 9855889 TI - Transitioning the critical care nurse from ICU to high-tech homecare. AB - Critical care nurses are in a good position to take advantage of the changes in health care. They can build a career that will be on the cutting edge of the changes in technology and care delivery if they are willing to learn some new skills and practice in a more autonomous environment. Nurses who are new to the homecare setting need to know more about insurance systems, diagnosis coding, and outcomes tools and prepare themselves for the rapid growth of computer assisted technologies that are being introduced into the practice of homecare. They also need to take the whole continuum of care into consideration when planning care for patients in the home and to be aware of community and family supports that can enhance the patient's health related quality of life. PMID- 9855890 TI - Care of the critically ill client at home. AB - The high-tech homecare nurse is operationalizing the trend to bring critical care nursing and medical services directly to the homecare environment. With the continuing advancement in technologic capability, the role of this newly emerging critical care nurse clinician will grow in scope of practice. As the health care industry strives to create more cost-effective solutions to the nation's burden of health care delivery, the relocation of complex medical interventions to the homecare setting is one method of effecting a seamless health care delivery system, where quality of care is maintained while costs are controlled. PMID- 9855891 TI - Telecommunication technologies in high-tech homecare. AB - The future of critical care nursing will be dependent upon the subspecialty's ability to prepare the high-tech homecare nurse for practice in the next millennium. Client self-management of highly complex interventional therapies will be enabled by the high-tech homecare nurse, who will facilitate the transitioning of critical care nursing to the home setting. Rapidly evolving technologic capabilities will be managed by the high-tech homecare nurse, who will assist in the development of a seamless health care delivery system, where quality of care is maintained in settings beyond the traditional boundaries of the intensive care unit. PMID- 9855892 TI - High-tech homecare infusion therapies. AB - Dramatic changes in health care delivery have created an increased reliance on ambulatory care. Adults and children increasingly are receiving infusions of intravascular medication or fluid in the home for a wide array of conditions. Several therapies have been adapted to the home, including chemotherapy, BRMs, TPN, blood components, anti-infectives, analgesics, inotropic agents, and investigational drugs. High-technology therapy in the home mandates safe and reliable vascular access to maximize positive outcomes, independence, and self care; minimize costs and disruption of daily activities and work schedules; and enhance client and family adjustments and satisfaction. Currently, the four main VADs for vascular access are central venous catheters (CVCs), peripherally inserted central catheters (PICCs, implantable infusion ports, and external or implantable ambulatory infusion pumps. PMID- 9855893 TI - Homecare, technology, and the management of respiratory disease. AB - Taking care of ventilator-dependent patients in the home is demanding and complex. The difficulties that families face can be reduced by provision of homecare services that are the result of the collaborative work of a variety of health care disciplines. These difficulties cannot be eliminated, however. Sensitive care requires that the nursing case manager be aware of signs that indicate the costs are exceeding the families' ability and plan to meet their needs and the needs of the patients. PMID- 9855894 TI - Management of psychiatric symptoms in medically ill patients in the homecare setting. AB - Nurses with critical care knowledge and experience increasingly provide technologically advanced care to patients at home who often present comorbid psychiatric symptoms. Failure to accurately recognize and provide appropriate intervention can be devastating for patients because these symptoms can exacerbate the medical condition and may lead to mortality. The Biopsychosocial Model and the Transactional Model of Stress were presented as guides for nursing practice. Practice is directed at 1) understanding the patient's perception of events, 2) involving the caregiver(s) in the patient's recovery, and 3) enhancing the individual's adaptive coping efforts and resources. The article attempts to guide nurses in making decisions on when to consult and collaborate with an APPN to achieve desired patient outcomes. PMID- 9855895 TI - High-tech wound and ostomy care in the home setting. AB - The role of the home health care nurse has expanded as a result of a burgeoning client base. This client group is sicker and relies on technology at an ever increasing rate. As the critical care nurse transitions into the homecare setting, wound, ostomy, and fistula management will require a broader base of expertise. Adequate reimbursement is tied to accurate assessment and documentation. The clinician armed with these skills is better prepared to develop an effective plan of care for this group of clients. PMID- 9855896 TI - Pediatric considerations in homecare. AB - "If I had known beforehand how difficult, demanding, time consuming, and exhausting it would be--having my child home on a ventilator--I would never have agreed to bring her home" (personal communication with a parent, 1994). This mother's statement strikes at the heart of pediatric high-tech homecare. Parents assume caregiver roles that professional health providers have taken years to develop. Nurses, as strangers, intrude into intimate family relationships that have cultivated over years. Pioneering agencies attempt to fill a gap in pediatric care using guidelines that have been entrenched in the medical and economic models for years. The multiple dimensions of high-tech pediatric homecare require more than provision of technical nursing services. In homecare, nurses are challenged by cultural differences, language barriers, loss of control, family dynamics, practicing in unfamiliar environments, and new technology. To ensure quality nursing care, all professional dimensions need to be considered to be of equal importance. PMID- 9855897 TI - High-risk pregnancy and neonatal care. AB - The delivery of high-risk maternal and neonatal homecare has been shown to be a cost-effective model of care delivery. The homecare management of preterm labor, placenta previa. PIH, and hyperemesis gravidarum has proven to be a safe interventional strategy with successful clinical outcomes. Studies of home uterine activity monitoring and phototherapy have demonstrated that the use of advanced technologic devices is spearheading the drive to reduce hospital lengths of stay, enabling the safe return of the patient to the home setting, under the watchful supervision of the high-tech homecare nurse. High-risk mothers and their newborns can be assisted to achieve optimal clinical outcomes through a coordination of services provided by an interdisciplinary team of health professionals led by the homecare nurse. PMID- 9855898 TI - Diversity issues in the homecare setting. AB - As this nation becomes increasingly more culturally diverse and seeks to compete in a global health care market, it is incumbent upon the nursing profession to prepare high-tech homecare nurses who are experts in their ability to make appropriate culturologic assessments of the clients whom they provide care. The culturologically competent high-tech homecare nurse is one who is able to obtain value-free culturologic information and who utilizes that information in the development of culturally sensitive plans of care. This article has provided a framework, with supporting rationale, to assist the high-tech homecare nurse in the gathering of culture-specific data from clients whose care delivery occurs in the richly diverse home setting. PMID- 9855899 TI - Mobilization of the interdisciplinary critical care team at home and abroad. AB - Already it is clear that managed care organizations seek delivery innovations that can move homecare beyond being a posthospitalization add-on to becoming a true alternative to hospitalization. Throughout its history, home health care nurses have epitomized Florence Nightingale's philosophy that nurses are "messengers of health as well as ministers of disease." With the development of more sophisticated models of interdisciplinary practice in high-tech homecare, nurses are in a position to help create a health care delivery system that in a sense brings people back to the future. The natural setting of the home is the best place to provide holistic care, it is where patients overwhelmingly prefer to be cared for, and it is affordable. Indeed, affordability has become the operant word, worldwide, for a health care delivery system that can continue to meet the growing demands of the future. PMID- 9855901 TI - Future care needs. PMID- 9855900 TI - Gullible guinea pigs. PMID- 9855902 TI - Ageism real or imagined? PMID- 9855903 TI - Counselling skills for carers and early stage dementia care. PMID- 9855904 TI - A prompt to healthy eating. PMID- 9855905 TI - Pressure sore care. AB - This paper reports the conduct, results and implications of a survey of residential and nursing homes for elderly people in Liverpool, with particular reference to the prevalence and care of pressure sores. The objectives of the survey were to establish prevalence rates, to collect data on the process of preventing and managing pressure sores, and to look at differences between the two types of home sector in terms of care policy and practice. PMID- 9855907 TI - Who cares pays. PMID- 9855908 TI - Assessment works. PMID- 9855906 TI - The quality of care. PMID- 9855909 TI - An open book. PMID- 9855910 TI - Evidence from the Royal College of Nursing to the Royal Commission on Long-term Care. PMID- 9855911 TI - Using the Barthel index in care homes. PMID- 9855912 TI - Euthanasia: a pragmatic view. AB - There will never be a simple resolution of the euthanasia debate, and perhaps there should not be. However, any policy in a pluralistic society should be based on the wishes of the individual so long as those wishes do not adversely affect others. Perhaps, as in the Netherlands, euthanasia needs to be decriminalised without being made legal, in a similar way to the decriminalisation of soft drugs such as marijuana (interestingly, the Dutch laws on drugs have also been held up as a possible standard for the USA as drug use in the Netherlands is much less problematic than in the USA (36). Individuals should be able to make difficult decisions regarding their own lives and healthcare workers should have the freedom, without fear of prosecution, to allow people in their care to pursue those options with dignity. This article has supported the proponent view of euthanasia--that it can be beneficial as it affirms individual autonomy. Compassionate carers must consider all the arguments in order to determine their own personal stand on the issue. PMID- 9855913 TI - The right to grieve. PMID- 9855914 TI - A ripe old age. PMID- 9855915 TI - Promoting urinary continence in residential care. PMID- 9855916 TI - Breaking down barriers. PMID- 9855917 TI - Education and training. PMID- 9855918 TI - Making ourselves heard. PMID- 9855919 TI - Change and challenge. PMID- 9855920 TI - Maximising life potential. PMID- 9855921 TI - Breaking the silence. PMID- 9855922 TI - Noteworthy benefits. PMID- 9855923 TI - Listening to Enid. AB - When the time came for Enid to make the transition into long term care the profile of Enid with her biography and behaviour modes was passed on to staff in the nursing home, thus enabling them to start from a point of knowing Enid as an individual. During her time at Burford, Enid managed to communicate and express herself sufficiently to keep a good degree of self-agency. Her dementia mapping scores showed her to be generally in a state of well-being. This article has attempted to show how it is possible to listen and learn from people with dementia and that care staff must want to learn about and from the person with dementia if they are to care for them in a person centred way. PMID- 9855924 TI - Homing in on quality. PMID- 9855925 TI - Laxatives and faecal incontinence in long-term care. PMID- 9855927 TI - Parkinson's disease. PMID- 9855926 TI - Constipation. PMID- 9855928 TI - Keeping the record straight. PMID- 9855929 TI - The need for single registration care homes: the RCN vision. PMID- 9855930 TI - Continuing the good work. PMID- 9855931 TI - Policy issues: your shout. PMID- 9855932 TI - Mental health assessment for older people. PMID- 9855933 TI - No care like home. PMID- 9855934 TI - Depression, anxiety and hallucinations in Parkinson's disease. PMID- 9855935 TI - Multidisciplinary approach to Parkinson's disease. PMID- 9855936 TI - Learning to implement OASIS. PMID- 9855937 TI - To tell the truth: the Joint Commission's sentinel event policy. PMID- 9855938 TI - The home care nurse as an informed voter. PMID- 9855939 TI - Home Healthcare Nurses Association's response to change: a convention report. PMID- 9855941 TI - The puzzle of Fran: home healthcare in a hurricane. AB - A natural disaster in the form of Hurricane Fran resulted not only in stories of ingenuity and compassion, but in a major performance improvement (PI) process for the entire agency. Through this PI process we learned about ourselves as a home health agency and discovered ways to improve our performance. More importantly we discovered ways to improve patient tracking and care during a disaster. PMID- 9855940 TI - Functional assessment of the home health client. AB - Since the collection of Medicare patient information through the use of the Outcomes and Assessment Information Set (OASIS) has become mandatory, it is increasingly important that home care nurses understand how to conduct a functional assessment. This article presents each element of function that should be assessed, explains how to assess each element, and describes some of the most common instruments used in performing a functional assessment. Additionally, tips are included for assessing the home environment and the client's safety. PMID- 9855942 TI - Assessing medication knowledge and practices of older adults. AB - An assessment instrument for home health nurses to use in assessing medication knowledge and practices of older adults was developed and tested on a convenience sample of 20 adults 65 and older admitted to a local home health agency. The tool was found usable by nurses, understood by patients, and had adequate test-retest reliability. The results emphasized the need for thorough medication assessments of all home health patients and provided a tool that home care nurses can use. PMID- 9855943 TI - Nursing across cultures: the Vietnamese client. AB - Asian Americans are the most rapidly growing immigrant group in the United States today. Home care nurses will be caring for increasing numbers of patients originating from the Southeast Asian peninsula in the countries of Vietnam, Cambodia, and Laos. A Vietnamese case study explores some of the beliefs, values, and practices held by this group of Asian Americans. The case study highlights the problems that can occur when a nurse is unaware of the impact that culture has on a client from a different ethnic and cultural background. Six steps to provide culturally competent care are outlined. PMID- 9855944 TI - Home nursing: a rural case study. PMID- 9855945 TI - The 24-hour day in home health. PMID- 9855946 TI - Effective use of a registered dietitian in home healthcare. PMID- 9855947 TI - Buzz me, beep me, page me, fax me! PMID- 9855948 TI - United we stand--divided we fall. PMID- 9855949 TI - Expert home care nurses provide guidelines that make a difference. PMID- 9855950 TI - An open letter to members of United States Congress. PMID- 9855951 TI - The physician as a member of the home healthcare team. AB - The physician is a vital member of the home healthcare team. Based on the Medicare legislation and regulations. The physician's participation is required for patient care and agency reimbursement. The home health care nurse is the important link between the patient and the physician. The compliance with, and sharing of, information related to the clinical, administrative, legislative and regulatory aspects of care benefits the patient, family, physician, nurse and the home health agency. PMID- 9855952 TI - The regulatory changes of 1998. PMID- 9855953 TI - Summary of the nursing practice guidelines for the cardiac home care patient. PMID- 9855954 TI - Using standards and guidelines in your daily practice. PMID- 9855955 TI - What every home healthcare nurse should know about complementary therapy. PMID- 9855956 TI - Current functional assessment tools. AB - In last month's issue of Home Healthcare Nurse, the article "Functional Assessment of the Home Health Client" described the value of performing functional assessment in relation to OASIS and methods to use in assessing a client's ability to perform activities of daily living (ADL) and instrumental activities of daily living (IADL). Although many different instruments are used to measure functional ability, this article will describe three of the commonly used instruments for measuring functional ability: the Barthel Index, the Functional Independence Measure (FIM), and PULSES have been tested for reliability and validity. The WeeFIM will also be discussed. PMID- 9855957 TI - Writing for publication: if I can do it, so can you! PMID- 9855958 TI - Four nurses share their experiences in cardiac home care. PMID- 9855959 TI - Special interest groups--shaping professional practice. PMID- 9855960 TI - Education is the key to understanding outcomes. PMID- 9855961 TI - Jumbo mambos. PMID- 9855962 TI - The challenge to humanity: perspective from South Africa. PMID- 9855963 TI - More warm soup, please. PMID- 9855964 TI - Blunt cardiac injury in the elderly trauma patient. AB - Elderly patients are at significant risk for development of complications after a traumatic event as a result of the aging process. This case discusses the diagnosis and sequelae of blunt cardiac injury, as well as the needs and approaches for an elderly patient. Nursing care interventions are provided. PMID- 9855965 TI - Bedside insertion of inferior vena cava filters. AB - Certain types of injuries make a patient more at risk for the development of a DVT or PE. Because the condition of such patients is critical, the treatment options are limited, and special attention is required so as not to cause further harm. Bedside procedures that preclude the need to transport a patient to another department offer clear advantages. By describing a tested clinical procedure- percutaneous insertion of the IVC filter--trauma nurses can gain a better understanding of how to prepare the patient, expedite the process, and provide ongoing nursing management. PMID- 9855966 TI - Internet access to nursing associations, Part 1: United States. PMID- 9855967 TI - FDA public health advisory: low molecular-weight heparin and spinal/epidural anesthesia or spinal puncture. PMID- 9855968 TI - RabAvert: new human rabies vaccine approved in the United States. PMID- 9855969 TI - A summer vacation. PMID- 9855970 TI - Multilingual health education booklets available on Website. PMID- 9855971 TI - To err is human ..., and very costly. PMID- 9855972 TI - Emergency department management of nerve agent exposure. AB - Nerve agents are toxic chemicals developed for use by the military, but used by terrorists against civilian populations. As threats of terrorism increase, it is possible that health care providers will be confronted with multiple victims of nerve agent exposure. Nerve agents are highly toxic forms of organophosphate poisons that potentially could cause harm to anyone who comes in contact. Emergency personnel need to be familiar with the agents, know how to prepare for encountering and treating victims, and know how to protect all people involved from further poisoning. PMID- 9855973 TI - Phenomenological study of the North Dakota flood experience and its impact on survivors' health. AB - Grand Forks, North Dakota, was devastated by an unprecedented flood, forcing the evacuation of an entire community of 50,000 residents in 72 hours. A qualitative descriptive study with use of phenomenological methods was conducted 6 months after the flooding. Its purpose was to gain a deeper understanding of the meaning of a traumatic flood experience and its influence on the health of the residents of Grand Forks. Data were generated through interviews, observations, and media reports. The sample included 3 female and 3 male residents of the city of Grand Forks, North Dakota, who ranged in age from 26 to 77 years. Five essential themes were identified: (1) shock and disbelief, (2) uncertainty, (3) grief and loss, (4) emotional exhaustion, and (5) hope and meaning. The participants' descriptions of emotional exhaustion were similar to those found with survivors of posttraumatic stress disorder. Older adults appeared to be especially vulnerable. Nurses who are aware of the effects of natural disasters, particularly flooding, may be better able to identify vulnerable populations and understand the health needs of other survivors. PMID- 9855974 TI - A fever workup guideline. AB - Fever is a common clinical problem in patients hospitalized with trauma. When an infection is suspected, it is important to identify the source of fever and begin definitive care before more severe morbidity occurs. An undirected approach with "pan cultures" has resulted in a high rate of negative cultures obtained and has increased patient care costs. By developing and using an evidence-based fever workup guideline, this institution was able to evaluate and identify infections and reduce costs. PMID- 9855975 TI - Practical tips for airway management in the injured child. PMID- 9855976 TI - Internet access to nursing and related health associations, Part 2: International. PMID- 9855977 TI - MedWatch program. PMID- 9855978 TI - Medication errors: the problem and its scope. AB - Nurses can take an important lead role in protecting their patients, their peers, and themselves from the unfortunate consequences of medication errors. Medication errors occurs for a variety of reasons, including inaccurate communications and deficits in knowledge and performance by and among all health care professionals. These problems and deficits need to be addressed. USP stresses the importance of focusing on the process or system in which the error occurred, rather than focusing blame on an individual. By acknowledging all actual and potential errors, and by consistently assessing and reassessing systems, nurses can help reduce medication errors and contribute significantly to improve patient safety. PMID- 9855979 TI - Six common misconceptions about vaccination--and how to respond to them. PMID- 9855980 TI - Interference between digital television and medical telemetry devices. PMID- 9855981 TI - A personal perspective: meaning is found in life. PMID- 9855982 TI - Trauma is a surgical disease: how does the perioperative nurse contribute to trauma patient care? PMID- 9855983 TI - Dog bites in children admitted to Pennsylvania trauma centers. AB - Dog bites are a major public health problem in the pediatric population, requiring emergency treatment for wound repair and possible hospitalization in a trauma center. Data from the Pennsylvania Trauma Outcome Study were analyzed, and the records of 183 pediatric dog bite patients from 1990 to 1995 were evaluated. Dog bites were found to constitute a very small proportion of the total pediatric admissions to Pennsylvania trauma centers; however, the findings were similar to other reported studies. PMID- 9855984 TI - A guideline for managing potential cervical spine injuries. PMID- 9855985 TI - Intranets and critical care. PMID- 9855986 TI - Reports of anesthesia-related adverse events. AB - A case report involving an anesthesia adverse event that was received in the FDA database was used to demonstrate postmarket reporting of medical devices by the manufacturers and the FDA. Even common, everyday adverse events are evaluated and used to improve the safety and efficacy of medical products. Instructions for accessing the FDA MedWatch forms and coding manual are provided, and readers are referred to additional resources on the FDA website. PMID- 9855987 TI - The year 2000 and biomedical equipment. PMID- 9855988 TI - Those closest to human suffering. PMID- 9855989 TI - The evaluation boom: is it new? PMID- 9855990 TI - Are we facing a 'post-antibiotic era'?--a review of the literature regarding antimicrobial drug resistance. AB - Since the introduction of antibiotics in the 1940s, antibiotic resistance has become an increasing problem. Today, multiple-antibiotic resistance is commonly associated with a number of clinically important pathogens and is therefore an important issue in clinical nursing practice. Epidemiological studies identify a number of important factors associated with increases in antimicrobial resistance. These include patterns of antimicrobial use, changes in medical and veterinary care and social practices affecting the transmission of microbes. Bacterial mechanisms of antibiotic resistance and the genetics of resistance-gene transfer are explored, with the intention of developing nurses' knowledge and understanding of control measures. PMID- 9855991 TI - Contradictions between perceptions and practices of caring in long-term care of elderly people. AB - Care for elderly people in nursing homes is a subject of increasing interest as populations around the world age. Numerous studies have been reported in relation to caring, how it is defined and what it means to nurses and clients in different contexts. Nursing is equated with caring and yet reports of nursing home care frequently cite such uncaring practices as neglect, fostered dependency, infantilization and depersonalization. The contradictions are explored with the aim of improving care of elderly people through greater understanding of how caring is perceived and practised. PMID- 9855992 TI - Researching children: some methodological and ethical considerations. AB - This paper examines some of the methodological issues and ethical considerations in researching children. Collecting data from children raises issues of informed consent, establishing rapport, and privacy and confidentiality. Children's consent to research remains a contentious issue that requires further deliberations. Researchers need to be flexible in their research with children and be cognisant of the strategies that can be used to enhance the quality of responses from children. PMID- 9855993 TI - Developing an audit tool for primary nursing. AB - Despite the proliferation of research on primary nursing, most studies have investigated the effects of primary nursing on quality of care, job satisfaction and collegial relationships. Few researchers have attempted to isolate the key dimensions of primary nursing in an observable and measurable form. The purpose of this exploratory study was to devise an audit tool for primary nursing that builds on Mead's research and incorporates views of patients/clients, relatives, nurses and other members of the multidisciplinary team. Preliminary findings suggest that the audit is a useful indicator of the extent to which the crucial elements of primary nursing are present or absent in any clinical setting. Furthermore it suggests that while many wards claim to practise primary nursing there is a considerable variation in the extent to which this is actually carried out. PMID- 9855994 TI - Developing the nursing care of breast cancer patients: an action research approach. AB - This study set out to develop the nursing care of breast cancer patients at the Central University Hospital in Turku, Finland, using the methods of action research and to evaluate the results of the development effort from the point of view of both patients and nursing staff. A theoretical model of the care of cancer patients was constructed on the basis of existing knowledge of how people react in stress and crisis situations or at different stages of a difficult illness. A specific care programme for breast cancer patients was also planned for the hospital's oncology and surgical units. Data were collected using questionnaires. Results indicated that our theoretical model of the care of cancer patients worked reasonably well. They also clearly highlighted the areas in which there is room for further improvement. Patients' experiences of the nursing staff were generally positive and the majority of the nursing staff also regarded the development project as a success. PMID- 9855995 TI - Priorities for mental health and learning disability nurse education in the UK: a case study. AB - The closure of psychiatric and mental handicap hospitals, the emphasis on community rather than hospital care and the distinction between health and social care for people in the community, with its funding implications, have demanded that mental health and learning disability nurses face the challenges of developing new skills and new ways of working. In this paper the findings of a study funded by the English National Board for Nursing, Midwifery and Health Visiting (ENB) are reported. This study explored the impact of these reforms on nurses and their practice, and identified the educational needs of mental health and learning disability nurses that have changed as a result. The study took a multiple-case study approach and involved interviews with 22 'key informants' and 88 mental health and 59 learning disability stakeholders in six sites, three in each nursing specialty. Inductive analysis of the interview data revealed 20 categories which summarize the major issues facing the two nursing specialties. These categories are discussed under seven themes and issues for nurse education are raised. PMID- 9855996 TI - Stroke patients' perceptions of hospital nursing care. AB - While nursing remains uncertain about the nature of its role in stroke rehabilitation, recent research by social scientists re-affirms its importance. This study explored a small number of stroke patients' perceptions of hospital nursing care in order to illuminate the meaning of the role in practice. Informants' early experiences of nursing care in general medical and care of the elderly assessment wards were dominated by having the necessary done, while nursing in the Stroke Rehabilitation Unit was characterized by a do it yourself approach. These two concepts are explored in detail and related to the existing literature, suggesting some specific areas for practice development and research. PMID- 9855997 TI - Using the Delphi technique to identify the content and context of nurses' continuing professional development needs. AB - In order to identify and prioritize the development needs of medical and surgical nurses, their possible approaches to learning, and the contextual factors influencing their professional development, a Delphi survey was carried out using a panel of 28 participants. Current development needs included skills in essential clinical care, specialist nursing, changing roles, patient care management, research and practice development. Although there was ambivalence about changing nurses' clinical roles, participants expected roles to diversify in the future. The need for responsiveness to change was emphasized, as was the need to maintain current areas of expertise. Professional development activity could take many forms, but fostering an organizational climate in which development was inherent in everyday working practices was felt to be as valuable as formal course attendance. Restricting factors included lack of time, resources, support and recognition. The survey has enabled professional and practice development activities to be prioritized locally, and the method used could be readily applied to other settings. PMID- 9855998 TI - Time in hospital. AB - This article is based on research into patients' perceptions of being in hospital. Time, ordinarily taken for granted, becomes a significant problem for people in hospital. Patients seek control over time that they do not seem to have during their stay in hospital. They try to keep track of time and find ways of passing it, while believing that precious personal time is lost to them. The insight gained into differing perceptions of time and how patients coped with these can be used to reduce some of the tensions and negative aspects of the way time is experienced in hospital. PMID- 9855999 TI - Facing uncertainty and possible death: the Christian patients' experience. AB - This paper presents the hospital experience of a group of coronary artery bypass graft surgery patients. A description of the qualitative methodology is given. The participants' experience was identified as facing uncertainty and possible death. Excerpts from interviews are given. The participants coped with the situation by praying or having others pray on their behalf. Their uncertainty and possible death was affected by the conditions of: available support; environment; credible authority; information; belief; and their psychophysiological status at the time. PMID- 9856000 TI - The effect of pre-operative information on post-operative anxiety, satisfaction with information, and demand for analgesia in Chinese men having transurethral resection of the prostate (TURP). PMID- 9856001 TI - Alteration without change?: a commentary on the proposed policy reforms of the British National Health Service. PMID- 9856002 TI - The NHS as a learning organization: aspirations beyond the rainbow? AB - AIM: It is the intention of this paper to review the issues and challenges organizations face when aspiring to embrace and enact the tenets of a learning organization; and in particular the perceived impact on management strategy, structure and leadership styles. The paper is predicated on the premise that learning and knowledge act as vital strategic resources, crucial not only to organizations in terms of competitive advantage but to ethical enterprise per se. BACKGROUND: Modern life is characterized by change, against the backdrop of this continual turmoil, organizational learning has emerged as a dominant theme within contemporary management theory, with many commentators increasingly locating the capacity of an aspiring organization to accommodate the ethos of organizational learning, as the vital component in ensuring enduring efficiency, innovation and competitiveness. However, the utility of such learning needs to be scrutinized and evaluated in terms of service need and expectation. ORIGINS OF INFORMATION: The paper will expand upon wider theoretical debates extant within the literature, by considering the concept and utility of the learning organization with specific reference to management reform extant within the British National Health Service (NHS). DATA ANALYSIS: During the course of the review the various the theoretical positions contributing to the notion of the learning organization will be analysed, the practical ramifications of which will be examined in the context of reflective practice, clinical supervision and the wider cultural background of nursing and the NHS. CONCLUSIONS: The paper concludes that the NHS needs to reorientate management perspectives to focus attention more acutely on systems which are deliberately designed to facilitate shared learning, to unravel the ambiguities of organizational life, to affirm management belief in the nursing contribution and to achieve an as yet unrealized potential in terms of patient care and advanced nursing practice. PMID- 9856003 TI - Power, professions and evidence-based guidelines: lessons from an organizational case study. AB - AIM: This paper addresses the issues of evidence-based practice (EBP) in relation to power, professionalism and the British National Health Service reforms. BACKGROUND: There has been little discussion at organizational level over issues of professional boundaries, roles and tensions, which are inextricably linked with the drive for an evidence-based culture. METHODS: The paper is a reflection on the findings of a recent case study, which examined the management and organization of an independent sector provider of mental health services in its drive to address issues of EBP. FINDINGS: New managerial hegemony had altered the balance of power within the organization, there was an extension of managerial surveillance, competing political relationships and a general decline in medical dominance. CONCLUSION: Organizations should avoid 'personalizing' the debate on establishing a research-based culture. EBP requires structures to support it and groups representing the health care professions within the organization should be involved in identifying and making recommendations on EBP issues. PMID- 9856004 TI - Merger management: a challenge to nursing leadership. AB - AIM: The article reviews the merger process of two obstetric divisions. BACKGROUND: Mergers of acute care facilities are becoming common due to the need to move towards a market orientation. There is a growing emphasis on corporations, competition, and profit and cost control. Nursing leadership in managing merger processes is crucial. METHODS: Information originated from administrative decisions, feelings expressed by the staff, author observations, and statistical data. Data was analysed by illustrating the merger phases, compared to the literature and research studies. KEY ISSUES: Issues related to planning and preparing the process, the management of human resources, the development of organizational culture, and the physical changes, are vital. CONCLUSIONS: Planning ahead, involving all partners from the early stages, extensive dialogue among colleagues and strong nursing leadership are key elements for a smooth transition. PMID- 9856005 TI - Mental health services--maintaining strategic direction. AB - AIM AND KEY ISSUES: This article reviews the theoretical basis of strategic management in an attempt to provide managers with a better understanding of the underpinning concepts and consequent actions they need to take to avoid loss of control and ultimate failure. BACKGROUND: The authors argue that community care for the severely mentally ill is failing and that in part the reason for this perceived failure is a closer allegiance to primary care that has shifted the focus away from mental illness. Such a shift, coupled with poor management and a desire by Community Mental Health Nurses (CMHNs) to retain a broad focus and maintain the 'autonomy' they gain when not held in the gravitational grasp of Consultant Psychiatrists, has resulted in strategic drift. The authors suggest 10 possible reasons to explain why CMHNs currently fail to meet the needs of the severely mentally ill. Among these are a lack of explicit strategic implementation plans, professional ambivalence and self-interest, poor management of resources and conflicting demands from key interest groups. CONCLUSION: It seems that mental health services in this country have reached the point where resistance to change should be crumbling in the face of perceived failure to deliver the required services. Whilst it could be argued that a major and potent source of internal change is performance gaps, few things force change more than sudden and unexpected information about poor organizational performance. PMID- 9856006 TI - The processes of case management: a review of the evaluation of a pilot study for elderly people in Hong Kong. AB - AIM: This paper is based on research into case management that aimed to evaluate the processes of the introduction of case management for elderly people into the community nursing services in Hong Kong. BACKGROUND: The Hospital Authority in Hong Kong introduced a pilot Case Management scheme into the Community Nursing Services. A research project was therefore developed to evaluate this case management model. METHOD: The processes were measured through information gained from group interviews, daily diaries and weekly activity sheets. FINDINGS: The roles and work practices of the Care Coordinators and Case Managers are described and the benefits of case management to patients, carers and nurses are highlighted. Difficulties are also discussed and good practices are identified. CONCLUSION: Staff in the hospital and community need time to get used to the model of case management and to the accompanying documentation. The role of the case manager needs to be clearly dilineated. PMID- 9856007 TI - Care pathways: have they a place in 'the new National Health Service'? AB - AIM: To discuss the use of Integrated Care Pathways (ICPs) as tools for ensuring cost-effectiveness and high quality patient-focused care. To examine how electronic pathways combining process, practice and audit might be effectively used by Primary Care Groups as a tool to integrate care within the 'New National Health Service'. KEY ISSUES: It has been argued that for doctors pathways lead to 'cook book medicine' and for nurses they move practice away from patient-centred care back to a disease/task-based model. This article explores the origins of pathways, addresses the concerns of critics and goes on to describe a pathway strategy. CONCLUSIONS: Pathway development offers the potential to create a coherent plan of care and treatment (across primary/secondary care) which incorporates evidence-based practice and reduces fragmentation and duplication whilst aiding co-ordination and communication. The incorporation of clinical indicators and measurable goals ensures the audit process is embedded. PMID- 9856008 TI - Assessing the new public management: the case of the National Health Service. AB - AIM: This paper reviews the origins, core principles and impact of New Public Management (NPM) in the UK public services, with particular attention paid to the emergence of NPM in the National Health Service (NHS). BACKGROUND: NPM represents a distinct break from traditional forms of public sector administration. Whilst its origins are grounded in economic crisis and New Right ideology, NPM philosophies and practices have since found wider acceptance. ORIGINS OF INFORMATION: Selective review of key journal papers, books and policy documents. KEY ISSUES: NPM is characterized by: managers being 'free to manage'; public sector disaggregation; competition; concern for efficiency; concern for outcomes; concern for quality; individualism rather than collectivism; and identification of the public service user as 'customer', 'consumer' or 'citizen'. CONCLUSIONS: NPM has taken root in the NHS. Future reform of the health service is likely to modify some aspects of NPM, though many of the core themes of NPM in the NHS look likely to remain for the foreseeable future. PMID- 9856009 TI - Culturally competent health care: a challenge for nurses in Saudi Arabia. AB - An emerging global reality is the presence of a culturally diverse workforce in health care environments. Not only are nurses delivering care to culturally diverse clients, the nurses themselves may come from different cultural and educational backgrounds. In order to provide culturally competent health care, such diversity must be molded into professional collaboration and respect. To date, health care to the citizens of Saudi Arabia has been provided largely by a foreign labor force, the vast majority of whom are non-Arabic speaking. This paper explores a number of strategies aimed at educating expatriate non-Saudi health professionals in the provision of culturally competent and culturally congruent health care. Leininger's transcultural health care theory is identified as the framework for achieving this goal. The theory has proved to be of great relevance in a hospital setting where some 40 different nationalities are represented among the work force. In such a setting, the potential for cultural conflict and stress is very real. Health professionals new to the Kingdom are introduced to the cultural dimensions of health care as it relates to patient care modes, as well as living and working together in a transcultural environment. Transcultural nursing is a major component of a professional nurse practice model which provides a visionary perspective for nursing care. Within the practice model, transcultural care principles are used to guide education, clinical practice and nursing research. Furthermore, Leininger's theory serves to grasp a comprehensive view of generic (folk) and professional health systems and to identify ethical issues confronted by nurses in the transcultural setting. PMID- 9856010 TI - Transcultural nursing practice described by registered nurses and baccalaureate nursing students. AB - Using Leininger's Theory of Culture Care Diversity and Universality as a framework, this research examined transcultural practices of nurses and students. A survey was administered to a convenience sample of registered nurses and senior baccalaureate students with 767 usable questionnaires returned. Neither group expressed confidence in their ability to care for culturally-diverse patients. Registered nurses (RNs) reported assessing cultural factors and modifying practices more frequently than did students. Respondents reported their beliefs about transcultural nursing were influenced by being with people of other cultures, their own personal values, and education. Analysis of the open-ended questions revealed two major themes. First, both nurses and students perceive an overwhelming need for transcultural nursing. Second, nurses and students respond to cultural challenges by modifying their care. Modifications are based on language and communication, pain perception and relief, religious and spiritual dimensions, gender and family roles, and other values. Results suggest that nurses and students are aware of culture, recognize that culture influences the care they provide, and modify their health teaching and nursing care based on culture. The use of a conceptual framework to help make modifications in care was not mentioned. PMID- 9856011 TI - Nurse anesthetists' perspectives on improving the anesthesia care of culturally diverse patients. AB - This paper reports the findings from a study which investigated whether the personal experiences of graduate and student nurse anesthetists could provide information on ways to improve the anesthesia care of culturally diverse patients. The study was designed as an exploratory descriptive qualitative investigation involving interviews with informants who were nurse anesthetists from cultural groups other than the dominant group of Anglo-Americans in a large midwestern city in the United States (US). Information emerged from the data that provided pertinent considerations for the provision of culturally congruent nursing care. PMID- 9856012 TI - The experiences of Mexican Americans receiving professional nursing care: an ethnonursing study. AB - The focus of this study was to discover the experiences of Mexican Americans receiving professional nursing care. The purpose of this study was to explore the care experiences, views, patterns, and meanings of Mexican Americans who have received professional nursing care. The need for this study arises from nursing's lack of knowledge regarding the experiences and cultural values of Mexican Americans receiving professional nursing care and how these values influence the health and/or well being of Mexican Americans. PMID- 9856013 TI - Special research report: dominant culture care (EMIC) meanings and practice findings from Leininger's theory. PMID- 9856014 TI - Nursing education exchanges: concerns and benefits. PMID- 9856015 TI - Transcultural nursing experts/consultants. PMID- 9856017 TI - Policy statements to guide transcultural nursing standards and practices. The Transcultural Nursing Society. PMID- 9856016 TI - Twenty five years of knowledge and practice development transcultural nursing society annual research conferences. PMID- 9856018 TI - Current developments in midwifery in the USA. PMID- 9856019 TI - Quality of care in reproductive health programmes: education for quality improvement. AB - The provision of high quality maternity care will make the difference between life and death or lifelong maiming for millions of pregnant women. Barriers preventing access to affordable, appropriate, acceptable and effective services, and lack of facilities providing high quality obstetric care result in about 1600 maternal deaths every day. Education in its broadest sense is required at all levels and sectors of society to enhance policy formulation that will strengthen programme commitment, improve services with a culturally sensitive approach and ensure appropriate delegation of responsibility to health staff at peripheral levels. This paper is the second in series of three which addresses quality of care. The first (Kwast 1998) contains an overview of concepts, assessments, barriers and improvements of quality of care. The third article will describe selected aspects of monitoring and evaluation of quality of care. PMID- 9856020 TI - Women-with-midwives-with-women: a model of interdependence. AB - AIM: To develop a research-based conceptual model of midwifery practice. DESIGN: Qualitative using grounded theory. SETTING: Midwifery practice in New Zealand and Scotland. PARTICIPANTS: 250 midwives and 219 clients from the two countries. FINDINGS: Data from the participants provided the contextual background as well as the major categories. Forming the background were the concepts of: beliefs, colleagues, culture, experience, expertise, education, environment, friends, families, intuition, knowledge and professionalism. The major categories of the model which emerged from the data were: attending and presencing, supplementing and complementing, reflection and reflexivity. These are discussed as pairs while the basic social process (core category) of reciprocity embraces the whole midwife-client relationship. CONCLUSIONS: This model offers the beginnings of documentary evidence of the essence of the midwife-client relationship and may go some way towards the creation of a written body of midwifery knowledge. It has potential applicability for both midwifery education and practice. PMID- 9856021 TI - Support, sensitivity, satisfaction: Filipino, Turkish and Vietnamese women's experiences of postnatal hospital stay. AB - OBJECTIVE: To assess Filipino, Turkish and Vietnamese women's views about their care during the postnatal hospital stay. DESIGN: Interviews were conducted with recent mothers in the language of the women's choice, 6-9 months after birth, by three bilingual interviewers. PARTICIPANTS: Three hundred and eighteen women born in the Philippines (107), Turkey (107) and Vietnam (104) who had migrated to Australia. SETTING: Women were recruited from the postnatal wards of three maternity teaching hospitals in Melbourne, Australia, and interviewed at home. FINDINGS: Overall satisfaction with care was low, and one in three women left hospital feeling that they required more support and assistance with both baby care and their own personal needs. The method of baby feeding varied between the groups, with women giving some insight into the reason for their choice. A significant minority wanted more help with feeding, irrespective of the method. The need for rest was a recurrent theme, with women stating that staff's attitudes to individual preferences, coupled with lack of assistance, made this difficult. The majority of comments women made regarding their postnatal stay focused on the attitude and behaviour of staff and about routine aspects of care. Issues related to culture and cultural practices were not of primary concern to women. CONCLUSION: Maternity services need to consider ways in which care can focus on the individual needs and preferences of women. PMID- 9856022 TI - Design and methodology in a community, practice-based research network: a study of nurse-midwifery home-birth practice. AB - OBJECTIVE: To assess the feasibility, probable success, and inherent problems of conducting a multi-site, multi-year cohort study of planned home birth in the USA. DESIGN: A prospective cohort study. SETTING: 14 states across the USA, representing a variety of regions and settings. PARTICIPANTS: 29 home-birth midwifery practices, representing 44 individual midwives in both large and small, solo and group practices. DATA COLLECTION: Birth outcomes for all women initially enrolled in these home-birth practices. KEY CONCLUSION: Multi-site studies of evaluating outcomes in small community-based practices are possible if several requirements are met. IMPLICATIONS FOR PRACTICE: Understudied birth alternatives can be formally researched within a co-operative network of participating practitioners. PMID- 9856023 TI - A six-year retrospective analysis of shoulder dystocia and delivery of the shoulders. AB - OBJECTIVE: To investigate the diagnosis, incidence and management of shoulder dystocia. DESIGN: Retrospective review of all the deliveries between 1988 and 1993 which were coded on the computer data base as 'moderate difficulty with the shoulders' or 'shoulder dystocia'. SETTING: A teaching hospital in the south of England. PARTICIPANTS: The study population consisted of 15,658 women who had cephalic vaginal deliveries of babies weighing > 2.5 kg. The sample consisted of the 257 women reported to have moderate difficulty with the shoulders or shoulder dystocia at delivery. FINDINGS: There was a significant fall in the reported incidence of shoulder dystocia during the period under investigation. Eighty-six (1.1%) of the deliveries between 1988 and 1990 were reported to have been complicated by shoulder dystocia. In 1991 a second option was introduced to allow both shoulder dystocia or moderate difficulties with the shoulders to be coded after delivery. Following this change in categorisation, the incidence of shoulder dystocia was reduced to 30 (0.6%) reported cases in 1992-1993. Over the same period the reported incidence of moderate difficulty rose significantly from 29 (1.1%) in 1992 to 60 (2.4%) in 1993 (P < 0.001). Disparity was noted between the experience of practitioners and contemporary definitions of shoulder dystocia and this was highlighted by the number of reported cases of moderate difficulty. In this study practitioners used the term shoulder dystocia in a general sense to describe a range of difficulties encountered with the delivery of the shoulders and they identified many varied manifestations. In comparison, contemporary literature describes shoulder dystocia as a discrete entity. KEY CONCLUSIONS: Shoulder dystocia is a complex clinical scenario and perceptions of the incidence may be influenced by alterations in the delivery technique and changes in the diagnosis and documentation. IMPLICATIONS FOR PRACTICE: The application of traction may interfere with the normal mechanisms of labour and the spontaneous birth of the shoulders, whilst increasing the risk of trauma to the baby. It may be unnecessary to use such interventions routinely in the care of normal labouring women. PMID- 9856024 TI - Differences in midwives' approaches to pain relief in labour. AB - OBJECTIVE: To examine the influence of midwives' approaches on the care given to women for pain relief during labour. DESIGN: Non-participant observation was used to collect data on midwives' behaviours during interactions with women in labour. SETTING: A large teaching hospital in Northern Ireland. PARTICIPANTS: Observations focused on interactions that were related to pain relief and involved 11 midwives caring for 15 women throughout the first stage of labour. KEY FINDINGS: Midwives' approaches to pain relief differed; at one extreme the midwife acted as 'a cold professional' whilst at the other extreme she was observed to be 'a disorganised carer'. In the middle of these two extremes was the 'warm professional'; her approach had a positive influence on the women's experience of labour pain. IMPLICATION FOR RESEARCH: Further research is needed to develop a profile of the factors that 'make' for a warm professional. The information gained could be used to teach midwives the skills of being 'a warm professional', in particular, the value of effective communication for sensitive care such as pain relief in labour. PMID- 9856025 TI - A reappraisal of Simpson's introduction of chloroform. AB - OBJECTIVE: To examine the introduction of the use of chloroform in childbirth, assessing the role of Sir James Young Simpson, the various motives for the innovation, and the health and sociological consequences for those involved in childbirth. DESIGN: A review of the attitudes of mothers, midwives, obstetricians and family practitioners, based on contemporary publications and subsequent historical analyses. FINDINGS: The context in which chloroform was introduced was fundamentally important to its widespread acceptance in the UK. The use of this drug carried serious implications for the childbearing woman, as well as the practice and status of those who provided her care. KEY CONCLUSIONS: The implications of the introduction of chloroform in childbirth were short, medium and long term. The changes associated with the acceptance of this drug may bear comparison with those relating to certain, current, pain-control methods. PMID- 9856026 TI - Little things mean a lot. PMID- 9856027 TI - Treating an arterial ulcer. PMID- 9856028 TI - When medications aren't given. PMID- 9856029 TI - Controlling bone pain. How to keep nausea and oversedation out of the picture. PMID- 9856030 TI - Myths & facts ... about phlebotomy. PMID- 9856031 TI - Lipid-lowering drugs and headache. PMID- 9856032 TI - Actionstat: autonomic dysreflexia. PMID- 9856033 TI - Treating HIV disease: hope on the horizon. PMID- 9856034 TI - Preventing chemical phlebitis. PMID- 9856035 TI - 11 easy ways to build rapport. PMID- 9856036 TI - Getting in touch with impaired foot sensitivity. PMID- 9856037 TI - Liability UAPs, and you. PMID- 9856038 TI - A short course lispro insulin on a quick performer. PMID- 9856039 TI - "Multiple accident victims, all elderly:" would our disaster plan be up to the challenge? PMID- 9856040 TI - Are you ready for a new position? PMID- 9856041 TI - Other places to explore. PMID- 9856042 TI - Beaming in on radiation therapy. PMID- 9856043 TI - Combating infection. Help from the web. PMID- 9856044 TI - Using therapeutic communication. PMID- 9856045 TI - Duty calls. PMID- 9856046 TI - Percutaneous sheath introducer. PMID- 9856047 TI - On my doorstep. PMID- 9856048 TI - Measuring critical care redesign: impact on satisfaction and quality. AB - This report describes restructuring on four critical care units at an acute, tertiary care hospital in South-east Pennsylvania. Utilizing a Patient Centered Care conceptual framework that had been successfully applied in the medical surgical areas, restructuring involved three main areas: revamping of work processes, inclusive of redesigned staff roles; environmental and facility changes; and enhancement of telecommunication and information systems. Preliminary analyses six months post redesign revealed improvements and maintenance in four outcomes areas--satisfaction, quality and efficiency, and costs of care. PMID- 9856049 TI - Managing organizational challenge and change: closing an inpatient unit. AB - The effects of a turbulent health care delivery market have impacted the day-to day reality of acute care hospitals. One effect is that the supply of acute care hospital beds currently exceeds the demand, a trend that is expected to continue to the year 2000 and beyond. Nursing administrators at St. Marys Hospital Medical Center made the decision to close an inpatient unit in order to better match acute care bed supply to existing demand. Decision support for closure, organizational change, and lessons learned from the closure process are discussed. PMID- 9856050 TI - Nursing competence--an evolving regulatory issue in Kentucky. AB - Competency validation for nursing licensure is an evolving regulatory issue in Kentucky. Societal and professional forces are driving the shift from mandatory continuing education requirements for relicensure to a competency-based evaluation paradigm. The Kentucky Board of Nursing has issued a working position paper, Accountability and Responsibility of Licensees to Assure Nursing Competence (October 1997), as the basis for a dialogue about shaping Kentucky nursing regulation in the twenty-first century. While the position paper explores new models for both initial and continuing licensure requirements, this article focuses on continued competence issue. PMID- 9856051 TI - Becoming involved: the nurse leader's role in encouraging teamwork. AB - This study explored why nurses enjoy and seek involvement in their work. Nurses from inpatient and outpatient units were surveyed. Results indicated that prior involvement, and meaningful involvement, served to predict further teamwork. Recommendations were made to the nurse leader who wished to promote increased work involvement. PMID- 9856052 TI - Transforming rural health care: Appalachian Regional Healthcare. AB - Appalachian Regional Healthcare is in the midst of transforming itself into a new organization in terms of culture, professional practice, and work processes. Just a few years ago, many members of its work force had never used computer technology. Today, nearly all employees, including nurses, have been trained in basic computer skills and are transitioning into an era when this rural integrated health care delivery system will employ a completely electronic medical record. PMID- 9856053 TI - Academia as partner in organizational change. AB - Partnerships are valuable strategies for promoting organizational change. Collaboration between academia and service can provide rapid access to new clinical service delivery mechanisms and real-world laboratories for implementing and testing novel approaches to care delivery. Academic-service partnerships also provide opportunities for work force development. One example of a partnership is described in this article. Details of a Nursing Care Management Institute illustrate principles of good practice for community-campus partnerships. PMID- 9856054 TI - Care tracker: a new approach to nursing care in ambulatory settings. AB - The shift from inpatient to ambulatory settings and the increasingly large and diverse numbers of ambulatory clients require reengineering and cost-effective approaches to managing ambulatory care. With data on 921 encounters in 12 ambulatory clinics, we developed CARE TRACKER, an instrument to describe nursing encounters by type, intervention, staff, time, and cost. Distinguishing nursing from medical interventions is an appreciable improvement over the traditional practice of bundling nursing as part of overhead costs. If managers can track the care delivered, they can better calculate staffing requirements, staff mix, inservice needs, and the costs of their existing and projected services. PMID- 9856055 TI - Partnerships in work redesign. AB - Nursing and other health care professionals, departments, and communities must integrate as partners for work redesign. Prescription refills and lab results are examples to exemplify partnerships and cost-effectiveness. Team strategies are discussed within the nursing functions of telephone triage and the coordination of patient flow, including implications for nurse administrators. These strategies can be adapted to a variety of health care settings. By working together as partners for work redesign, all members of the health care team become part of the gold standard for care delivery. PMID- 9856056 TI - Reengineering for a new millennium: the year 2000 problem in health care. AB - Reengineering and restructuring of systems--indeed, entire organizations--has become commonplace in today's volatile health care environment. Of all the reengineering to come, the most immediate is year 2000 compliance. This article will discuss the problem in general and its urgency, and why it's particularly critical that health care achieve compliance. In addition, it will discuss key issues to address in a conversion project, such as the importance of including clinicians on the project team. PMID- 9856057 TI - Demonstrating the benefits of case management. PMID- 9856058 TI - Effect of a clinical case manager/clinical nurse specialist on patients hospitalized with congestive heart failure. AB - The purpose of this study was to evaluate the effect of case management by a Clinical Case Manager/Clinical Nurse Specialist (CCM/CNS) on hospitalized length of stay and hospital charge throughout a 12-month period for patients with congestive heart failure. A total of 491 patients were discharged during 1997 with a diagnosis-related group code of 127. Of this number, 88 were case managed by a CCM/CNS. The remaining 403 received the usual management of their care. The group who were case managed by the CCM/CNS demonstrated significantly shorter length of stay (t = 5.40, P < 0.00) and lower hospital charges (t = 4.26, P < 0.00) than the patients with congestive heart failure who were not case managed. Secondary analysis indicated a significant interaction between case management and involvement of a cardiologist in the care of the patient. Patients whose care involved a cardiologist without case management by a CCM/CNS demonstrated significantly greater (alpha = 0.01) length of stay and hospital charges than patients who were case managed by a CCM/CNS or patients whose care did not involve a cardiologist. PMID- 9856059 TI - The art of nursing: the client-nurse relationship as a therapeutic tool. AB - The C-N relationship is the cornerstone and essence of mental health/psychiatric nursing. However, that process has the same potential for nursing in general. The C-N relationship is an interpersonal, interactive, and ongoing relationship set up to assist clients in the continuous evolution toward quality health and well being. Although this article focuses primarily on the mental health population, the value of a positive working C-N relationship is paramount in addressing all health care issues. The C-N relationship is, indeed, a therapeutic tool that can generate positive side effects, enhance treatment approaches, and greatly influence favorable health care outcomes. It is easy to apply, is flexible, and, when used effectively, can generate the power within health care situations to determine desired health care outcomes. PMID- 9856060 TI - Building systems to measure continuity of care. AB - Case management is the primary means to provide continuity of care across multiple institutions and practitioners. Traditional case management is becoming more complex, because in a managed care environment, financial decisions may be as important as clinical decisions. Decisions on extended treatment may have to be justified to the patient's insurance company and may have to be based on pre approved treatment criteria. If not properly authorized, treatment costs could be denied, leaving the patient with sizable medical bills and increased dissatisfaction. A good computerized case management system can capture data from disparate sources, integrate it into common databases, display it in a real-time fashion to case managers, report it using standard (or custom) reports or templates, and have the capability to trend and project future treatment. This article offers case managers practical advice on the options available for obtaining or customizing a case management system that will be appropriate for their needs in a managed care environment. PMID- 9856061 TI - Fast tracking in cardiac surgery: the St. Francis experience. PMID- 9856062 TI - Designing a system for ambulatory obstetric case management. AB - The authors describe the development of critical pathways for ambulatory obstetric case management. When case management was identified as needed, but published work in outpatient obstetrics could not be found, four nurses used this opportunity to design a cost-effective system leading to quality outcomes. The driving force was the need for a format that directed comprehensive consistent care delivered by a large multidisciplinary health care team. Design issues included capturing leading edge standards of care and user friendly formats for all caregivers. Throughout a period of 2 years, a trifold format was developed for all obstetric patients, and 15 bifold formats were developed for patients with specific high-risk diagnoses. The format design facilitated cost-effective quality care and is expected to improve patient outcomes. A research study has been initiated to measure effectiveness of the design. PMID- 9856063 TI - Case management: client and manager perspectives. PMID- 9856064 TI - Care coordination in managed care. Creating a quality continuum for high risk elderly patients. AB - The portion of the American population 65 years and older is growing rapidly, and the group 85 years and older is the largest growing segment of our population. The country's largest health maintenance organization, Kaiser Permanente, in Fontana, California, experienced a 9% growth in its membership between 1996 and 1997. Following the Kaiser national Model of Care recommendations, Fontana's Kaiser Permanente created their department of Extended Care Services and implemented programs that coordinated patient care across the continuum to meet the needs of its members who were 65 years and older. The programs included new member screening and orientation, telephonic care coordination, and a volunteer based care call program. They also developed a medical management model that included a consultative geriatric assessment clinic and a primary care clinic as well as provision of team-based care to members residing in skilled nursing facilities. PMID- 9856065 TI - Professional ethics: are we on the wrong track? AB - Are we on the wrong track, in terms of our expectations of a code of practice, professional ethics teaching or the wider field of moral philosophy, in our search for clear answers to the ethical problems that arise in clinical practice; or are we simply wrong in believing that there are always clear answers? This article examines a particular case, an account of which appeared in Nursing Standard at the end of 1996. The conclusion reached is that we are likely to be misguided in assuming that there are always clear, straightforward answers to the ethical problems of clinical practice. However, more than a fleeting acquaintance with the field of ethics may help us to begin to think with greater clarity about what we do and why we do it. The hope is that this might lead to considered, humane practice by design, with the critical understanding and support of our professional peers and employers. PMID- 9856066 TI - Maintaining integrity through clinical supervision. AB - This article suggests that there is a relationship between successfully maintaining integrity in nursing and the practical provision of opportunities for shared reflection offered by good clinical supervision. In order to establish this case, I will first give some definitions and then proceed to consider how these ideas relate conceptually. The article makes no attempt to offer empirical research as confirmation, but provides a conceptual and moral argument making use of anecdotes for purposes of clarification and illumination. It is the author's belief that, if it is understood and implemented properly, clinical supervision offers a radical challenge to nursing's existing culture. If nurses are interested in the survival of the profession, it is perhaps a challenge that has to be addressed. PMID- 9856067 TI - The role of nurses in euthanasia: a Dutch study. AB - What role do nurses play in euthanasia? How do they experience this role and what should be their ideal role? These are the questions of a study undertaken to gain insight into the role of nurses in euthanasia. Answers to these questions were derived from 20 semistructured in-depth interviews with nurses employed in a Dutch hospital. To make clear the role of nurses in euthanasia, the issue was split up into four phases: observation of a request for euthanasia; decision making; carrying out of the request; and aftercare. This article is a brief report on the most important results regarding these four phases. Special attention will be paid to nurses who have conscientious objections. To evaluate the study results, an unambiguous interpretation of the concept of 'euthanasia' is of most importance. For that reason, Dutch laws and other regulations concerning euthanasia will be explained. PMID- 9856068 TI - The utility of futility: the construction of bioethical problems. AB - The aim of this article is to analyse the contemporary 'futility discourse' from a constructivist perspective. I will argue that bioethics discourse typically disregards the context from which controversies emerge and the processes that inform and constrain such discourse. Constructivists have argued that scientific knowledge is expressive of the dominant paradigm within which a scientific community is working. I will outline an analysis of 'medical futility' as a construction of biomedical and bioethical communities (and their respective paradigms). I will trace the emergence and utilization of futility in the literature. My analysis of the context (i.e. the historical circumstances, the particular actors involved) within which the futility discourse emerged suggests that medical futility was constructed, in part, as a means of enhancing physician domination of a context wherein medical authority was threatened. The actors in this debate express widely divergent frameworks of 'the good', arguing from distinctive representations of moral agency. At times, this controversy has been argued from incommensurate moral horizons wherein the discussants debate incomparable problems. This discussion is related to a study of the 'practice' of futility in the clinical context. Further studies on the construction of bioethical problems are a necessary condition for supporting the truth claims of bioethical arguments. PMID- 9856069 TI - Children sold for transplants: medical and legal aspects. Amnesty International- Danish Working Group for Children. AB - Over the last few decades there has been a substantially higher percentage of successful organ transplants but also a significant imbalance between the demand for and the supply of organs, creating the basis for a highly profitable black market trade in human organs. Sometimes there are reports that children have been kidnapped, only to reappear later lacking one kidney, or that they simply disappear and are subsequently killed to have all their transplantable organs removed for profit. The European Union feels that there is a need for action and that it has a duty to act in this field, especially for ethical reasons. There is now established close co-operation between the various European transplant organizations. The legal protection of children with regard to organ transplantation is not specifically mentioned in the existing conventions because this issue was not foreseen at the time of their preparation. However, the issue is covered in a broader sense by more general provisions. There are endless rumours surrounding this area. Members of various organizations who travel in the suspected countries say that the trafficking in children who are sold for transplantation is well known, but it is too difficult and very dangerous to catch the people involved. We have to conclude that it has been impossible to prove or disprove the rumours, but they are consistent and we all, especially in the health care professions, have an obligation to be keenly aware of how and where organs are obtained. PMID- 9856070 TI - Intimacy--meeting needs and respecting privacy in the care of elderly people: what is a good moral attitude on the part of the nurse/carer? AB - This article explores notions of intimacy in the caring context. The aspects discussed are: privacy and intimacy; intimacy as emotional and/or physical closeness; intimacy as touch; sexual intimacy and normal ageing; sexual intimacy and patients suffering from dementia; and intimacy as trust. Examples are given and problems are identified, with reflection on the attitude and behaviour of the carer. It is suggested that when trying to make moral decisions in concrete situations it is imperative that the carer is aware of the values upon which his or her own thinking is based. It is argued that the guiding principle should be the moral assumption that the carer's responsibility can never be interpreted as a right to disregard the wishes of the patient. Hence, the key word in daily care is 'respect'. PMID- 9856071 TI - Health care, ethics and nursing in Bangladesh: a personal perspective. AB - Health care in Bangladesh is in a sad condition, with not enough doctors and nurses available to serve its people, but, even with this limited number of health care professionals, better care would be possible if greed for money and unaccountability to the people were controlled by the Government. Conditions for members of the nursing profession are not acceptable for those who are dedicated to serving the sick. Acknowledgement of nursing's professional dignity is almost completely absent. In addition, the salary earned is not enough to make a living. There are in existence professional associations who are struggling for the rights of the nursing community, although few concrete results have yet been seen. This article is written from the perspective of the author's position as a member of the Board and Treasurer of the International Association of Bioethics, and her interest in feminism and bioethics, which justifies her link with oppressed nurses (because most are women) and unethical practices in the nursing profession in Bangladesh. PMID- 9856072 TI - Court-ordered cesareans--choice or control? PMID- 9856073 TI - The art of nursing: aesthetics or praxis? A response to Steven Edwards, Louise de Raeve and Per Nortvedt, by Stan van Hooft. PMID- 9856075 TI - 6th International Conference of Anticancer Research. Kallithea, Halkidiki, Greece. October 21-25, 1998. Abstracts. PMID- 9856074 TI - Growth factor stimulation of bone healing. Effects on osteoblasts, osteomies, and implants fixation. AB - Bone tissue has been shown to contain numerous cell-to-cell signalling peptides called growth factors. These growth factors are thought to have important regulating effects for bone remodeling and bone healing, due to their potent effects on bone cell metabolism. In vivo studies over the last half decade have demonstrated that growth factors candidates for future clinical use in orthopedic surgery. In numerous clinical situations enhanced bone formation and bone healing could lead to improved results of surgical procedures. This thesis describes the most important bone growth factors and their actions in vitro and in vivo. In vitro investigations of growth factor effects on osteoblast chemotaxis and metabolism are described as well as in vivo studies with growth factor stimulation of fracture healing and bone healing to prosthetic-like implants. In vitro results: Several growth factors exhibited chemotactic effects towards human osteoblasts. TGF-beta 1 and PDGF-BB had the strongest chemotactic effects, whereas PDGF-AA, IGF-1, and IGF-2 had less but significant chemotactic effects towards human osteoblasts. TGF-beta 1 exhibited the highest chemotactic potency with maximal activity at 100 pg/mL, whereas the other growth factors had maximal effects at 10-100 ng/mL. BMP-2 was found to have chemotactic effects toward human osteoblasts, human bone marrow osteoprogenitor cells, and U2-OS osteosarcoma cells. BMP-4 and BMP-6 were without any chemotactic effects towards these celltypes. Human bone marrow osteoprogenitor cells were the most responsive celltype to BMP-2 stimulation. Growth factor combinations resulted in synergic stimulative effects on different metabolic functions on human osteoblasts. Combinations with TGF-beta 1 and PDGF-BB strongly stimulated proliferation and chemotaxis. Combinations with TGF-beta 1, PDGF-BB and BMP-2 strongly stimulated an osteoblast differentiation parameter (alkaline phosphatase activity). The different growth factor combinations had no effect on collagen synthesis in human osteoblasts. In vivo results: Continuous application of 1 and 10 micrograms natural TGF-beta to a plated tibial osteotomy in rabbits increased mechanical bending strength and callus formation at 6 weeks observation. Diaphyseal cortical bone remodeling was not affected by the local growth factor application. In a dog model with unloaded implants surrounded by a gap, 0.3 microgram rhTGF-beta 1 adsorbed to gritblasted tricalcium phosphate coated implants, was able to enhance mechanical fixation, bone ingrowth and gap bone formation. 3.0 micrograms rhTGF beta 1 had less but significant stimulative effect. In a weight-loaded model, 0.3 microgram rhTGF-beta 1, adsorbed to gritblasted tricalcium phosphate coated implants, was able to enhance bone ingrowth, without enhancement of mechanical fixation. In the unloaded model, 0.3 microgram rhTGF-beta 1, adsorbed to gritblasted hydroxyapatite coated implants, was able to enhance bone ingrowth, without enhancement of mechanical fixation. 3.0 micrograms rhTGF-beta 1 had no stimulative effects. The establishment of a biological implant fixation concept with growth factor absorbed to ceramic coatings of implants was successful. These data are promising for a possible future clinical usage of growth factors, especially for enhancement of bone healing to cementless prosthetic components. PMID- 9856076 TI - Molecular taxonomy of a new potyvirus isolated from chilli pepper in Thailand. AB - A virus isolated from chilli pepper plants in Kamphaengsaen, Nakorn Pathom, showing vein banding mottle symptoms was classified using sequence analysis and phylogeny of the coat protein gene and 3' noncoding region (3'NCR). This virus was found to be a typical potyvirus on the basis of particle morphology, biological properties and cytopathology. The 3'terminus region of the genome of 1,309 nucleotides, representing the viral coat protein gene and 3' NCR was cloned and sequenced. Nucleotide sequence analysis indicated that the 3' region of the viral genome had a poly A tail of at least 12 nucleotides, a noncoding region of 272 nucleotides, a coat protein gene of 864 nucleotides and 161 nucleotides representing the 3' terminus of the polymerase gene. The amino acid sequence of the coat protein was compared with those of 23 distinct potyviruses, and 63.1% shown to be the highest homology. However the 3' NCR had, at most, 29.7% random homology, thus indicating that this virus is a distinct species in the genus Potyvirus in the family Potyviridae. The result is well supported by previous studies on the biology and biochemical properties of this virus. PMID- 9856077 TI - Effect of cysteine substitutions on dimerization and interfragment linkage of human cytomegalovirus glycoprotein B (gp UL55). AB - Experiments were carried out to analyze the function of cysteine residues at amino acid positions 506 (cI), 550 (cII), 573 (cIII), and 610 (cIV), in dimerization and/or disulfide linkage of human cytomegalovirus (HCMV) glycoprotein B (gB). Single c-codons or pairs were substituted in the gB sequence of constructs which were used for transfection and selection of stable transfectants. Analysis of gB expression products revealed that single substitutions of cIII or cIV, but neither single nor double substitutions of cI or/and cII prevented gB dimerization. All substituted gB derivatives were, however, no longer processed by proteolytic cleavage. After deletion of the membrane anchor domain, correct proteolytic processing was again observed for anchorless gB forms. Substitutions of cI or cI/cII in secretory gB appeared to interfere with disulfide linkage between gB cleavage fragments. In the case of anchorless gB with substitutions of cII, cIII, or cIII/cIV, however, extracellular gB forms were not recovered. Using the Sindbis expression system recovery of all anchorless gB forms with cysteine substitutions was achieved. Analysis verified involvement of cI/II substitutions in intrachain disulfide linkage between cleavage fragments of HCMV gB. PMID- 9856079 TI - Endogenous protease-dependent replication of human influenza viruses in two MDCK cell lines. AB - Multi-cycle replication and plaque formation of influenza A and B viruses and cleavage activation of their hemagglutinin (HA) by an endogenous protease(s) were examined in two MDCK cell lines, MDCK(-) and MDCK(+). No exogenous trypsin was required for multi-cycle replication and plaque formation of all the influenza A viruses tested in the MDCK(+) cell, while those of the viruses in the MDCK(-) cell were completely trypsin-dependent. In both cell lines, on the other hand, influenza B viruses grew well in the absence of trypsin. The capability of multiple replication and plaque formation of the influenza viruses correlated with cleavage of the HA precursor (HA0) to HA1 and HA2, indicating that both cell lines express an HA activating endoprotease(s); that of the MDCK(+) cell activates the HA of influenza A and B viruses, and that of the MDCK(-) cell does only the HA of influenza B virus. Furthermore, the protease of the MDCK(+) cell was strongly suggested to be present on the cell surface and a serine protease. The MDCK(+) cell would be useful for isolation of influenza viruses from clinical specimens and for screening of protease inhibitors for anti-influenza virus drugs. PMID- 9856078 TI - The association of skin diseases with human herpesvirus 8 infection in HIV carriers. AB - The seroprevalence to Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus type 8 (HHV-8) was surveyed in human immunodeficiency virus type 1 (HIV-1) carriers with or without skin diseases, and also in HIV-1 negative individuals in Thailand. Using an immunofluorescence assay, the seropositive rates to lytic antigens of HHV-8 in HIV-1 carriers with or without skin diseases were 25% and 7.4%, respectively, but none of HIV-1 negative individuals had antibody. The seroprevalence to HHV-8 antigens was high in HIV positive individuals with low CD4/CD8 ratio, suggesting that HHV-8 is reactivated during the immunosuppressive state. Several polypeptides with apparent molecular weights of 34-38,000 and 40,000, which were specific to HHV-8, were identified by the immunoprecipitation test using the seropositive sera. Our results suggested that HHV-8 co-existed with HIV in HIV-1 carriers and the existence of HHV-8 may be associated with clinical features in the skin. PMID- 9856080 TI - Further characterization of HIV RNA synthesis early after cell-to-cell transmission infection. AB - Using a one-step model for cell-to-cell transmission of HIV infection we have identified two distinct phases of HIV RNA synthesis. The first phase (4 h-12 h p.i.) was marked by an increase in only the full-length 9 kb RNA, while the second phase (24 h p.i. onwards) comprised a significant increase in the levels of all three species of viral RNA. We now report that while the continual presence of actinomycin D at 50 micrograms/ml abolished all detectable viral nucleic acid synthesis when virus donor H3B cells were pre-treated with 50 micrograms/ml of actinomycin D (AmD), washed free of unbound drug (a procedure which inhibited > 99% of total cellular RNA transcription) then mixed with untreated recipient Hut 78 cells, normal amounts of full length linear unintegrated viral DNA were produced and the first phase of viral RNA transcription was unaffected. Similarly, when both the virus donor cells and recipient cells were arrested in the late G1 phase of the cell cycle by aphidicolin and then mixed, linear unintegrated viral DNA was the major viral DNA species produced. The appearance of circular viral DNA and progeny virus was inhibited, but the first phase of induced viral RNA synthesis was unaffected. When AZT was added at 2 h or 4 h after cell-cell mixing, the level of 9 kb RNA detected was significantly lower, corresponding to reduction in the level of viral DNA. These and previous results indicate that the template for the first phase of viral RNA synthesis was likely to be newly synthesized, linear unintegrated viral DNA and not pre-existing proviral DNA present in the donor cells. Taken together, our results suggest that there exists a yet to be fully characterized pathway of concurrent viral DNA and RNA synthesis early after cell to cell transmission of HIV infection. PMID- 9856081 TI - A nonstructural and antigenic glycoprotein is encoded by ORF3 of the IAF-Klop strain of porcine reproductive and respiratory syndrome virus. AB - Open reading frame 3 (ORF3) of the genome of porcine reproductive and respiratory syndrome virus (PRRSV), Quebec strain IAF-Klop, was reverse-transcribed and cloned into the procaryotic expression vector pGEX-4T-1, then subcloned into the eucaryotic expression vector pAdCMV5 which was used as a shuttle vector to generate a replication-defective recombinant adenovirus. The procaryotic GST-ORF3 recombinant fusion protein was used to raise a monospecific antiserum in rabbits. By Western-immunoblotting with PRRSV-infected cell extracts, the ORF3 encoded protein had an estimated molecular mass (M(r)) of 42 kDa, similar to that of the protein expressed by the adenovirus vector. Endoglycosidase F digestion showed that the ORF3 encoded protein occurs in an highly glycosylated form (GP3) in the infected MARC-145 cells. Pulse-chase and radioimmunoprecipitation experiments revealed that the GP3 protein was present in amounts equivalent to those of the N, M, and GP5 proteins in the infected cells, whereas no GP3 could be detected in purified virions. During the first 30 min of chase, the GP3 undergoes a gradual downward shift of its apparent M(r), thought to result from trimming of the mannose-rich glycan structures. Tested convalescent pig sera that were found to be seropositive to PRRSV by indirect immunofluorescence reacted positively with the recombinant GST-ORF3 fusion protein by immunoblotting. Data indicated that the ORF3 protein of the Quebec reference strain of PRRSV is a highly glycosylated and antigenic protein, which is nonstructural. PMID- 9856082 TI - Characterization of haemagglutinin-esterase protein (HE) of murine corona virus DVIM by monoclonal antibodies. AB - We analyzed the characteristics of seven monoclonal antibodies (mAbs) raised against purified HE (hemagglutinin-esterase) glycoprotein of the murine coronavirus DVIM (diarrhea virus of infant mice). Immunocrossreaction of these mAbs with JHM and/or MHV-S suggest that antigenic epitopes of HE of DVIM are similar to those of JHM and/or MHV-S. Four mAbs (1b4, 3a28, 4c19, 10b7), designated as group A mAbs, strongly inhibited both HA and AE activities. On the other hand, three mAbs (5a3, 6a6, 13a4), referred to as group B, had a comparatively weak HA inhibition activity. These results indicate that the antigenic epitopes of this glycoprotein can be classified into at least two groups and that the functional sites of HA and AE activities are similar but not identical. Neutralizing activity was shown in group A mAbs exclusively, suggesting that the ratio of HA and/or AE activities may play important roles in the cell fusion activity of DVIM-infected cells. PMID- 9856083 TI - Sequence comparison of the major capsid protein gene from 18 diverse iridoviruses. AB - Insect iridoviruses (IV) have been found on all continents of the world and in a broad range of insect hosts. The host range for a single strain can cross several insect orders. This along with a paucity of molecular information on all but a few members has led to confusion in the taxonomy and classification of these viruses and in the identification of potentially novel isolates. To address this problem consensus PCR primers were designed to amplify and sequence a 500 bp region of the major capsid protein (MCP) gene. PCR products were amplified from eighteen IVs belonging to the genus Iridovirus. No product was observed for the chloriridovirus IV3. Phylogenetic analysis of the partial MCP gene sequence showed that the iridovirus genus can be divided into three groups. These results support previous studies where a range of molecular techniques were used. Group I contained PjIV and IV31, group II contained IV6 (CIV), IV21, and IV28, and group III contained IV1 (TIV), IV2 (SIV), IV9 (WIV), IV10, IV16, (CzIV), IV18, IV22, IV23 (BbIV), IV24, IV29, IV30, AgIV and an undescribed weevil IV. There was no correlation of relatedness with host of isolation but there was some correlation with geographic region of isolation. Sequence analysis also raised issues concerning the purity of some virus stocks and supported the view that some isolates should be considered as variants of one virus species. PMID- 9856084 TI - Regulation of the Epstein-Barr viral immediate early BRLF1 promoter through a distal NF1 site. AB - The immediate early BRLF1 and BZLF1 promoters of Epstein-Barr virus are crucial for triggering the replicative cycle of the virus. To better understand the cell type dependence of the lytic cycle we conducted an analysis of the BRLF1-promoter in the epithelial cell line HeLa and the lymphoid cell line IM9. To analyze promoter activities, transient transfections with 5'-deletions of the BRLF1 promoter in front of luciferase as reporter gene were conducted. Besides the already known cis-acting elements of the promoter close to the TATA-box, more distal elements were located and functionally tested. A nuclear factor 1 consensus site was found to act positively in HeLa cells, but did not in lymphoid IM9 cells. The NF1 site was shown to bind protein by electrophoretic mobility shift assays, antibody-supershifts and in vitro footprinting. Thus, a protein belonging to the nuclear factor 1 family of proteins was identified as additional cellular trans-acting factor for the BRLF1-promoter besides the already described factors Sp1, Zta and Zif268. PMID- 9856085 TI - Poor induction of interferon-induced 2',5'-oligoadenylate synthetase (2-5 AS) in cells persistently infected with mumps virus is caused by decrease of STAT-1 alpha. AB - Poor induction of interferon-induced 2',5'-oligoadenylate synthetase (2-5AS) has been demonstrated in cells persistently infected with mumps virus as compared with uninfected cells. As for the number of interferon (IFN) receptors and the level of IFN regulatory factors (IRF-1 and IRF-2) mRNAs, there was little difference between them. Therefore, it is suggested that the IFN-alpha signaling system is ineffective in the persistently infected cells. Components of IFN stimulating gene factor 3 alpha (ISGF-3 alpha), STAT-1 alpha (p91) and STAT-2 (p113), were investigated in human amnion (FL), human nasopharyngeal cancer (KB), human T-lymphoid (HUT 78), and human B-lymphoid (Akata) cells persistently infected with mumps virus. STAT-1 alpha, but not STAT-2, disappeared in these persistently infected cells, and this factor was not restored by treatment of these cells with IFN. However, no difference was observed between the levels of STAT-1 alpha mRNA transcript in persistently infected and uninfected control cells. It is reasonable to infer that the poor induction of 2-5AS activity is due to the decrease of STAT-1 alpha in correlation with the IFN-signal transduction pathway. Furthermore, induction of other IFN-stimulated genes (ds-RNA activated protein kinase, PKR, and MxA protein) was also reduced in the cells persistently infected with mumps virus. PMID- 9856086 TI - Nucleotide sequence of the 5'-terminus of Newcastle disease virus and assembly of the complete genomic sequence: agreement with the "rule of six". AB - We have determined the sequences of the 5' ends of three strains of Newcastle disease virus, permitting the assembly of the entire genomic sequence, which amounts to 15,186 nucleotides. This length is in agreement with the rule of six, which has been shown to determine replication efficiency in similar viruses. Comparison of the extreme 5' end of the trailer sequence with that of the 3' terminal leader sequence of the virus reveals a high degree of complementarity. Variation between the 5'-terminal sequences of the different strains reveals the presence of alternative L gene polyadenylation signals, leading to correspondingly different trailer lengths. PMID- 9856087 TI - Upregulation of HIV-1 replication in chronically infected cells by ingenol derivatives. AB - We have previously reported that ingenol derivatives are highly potent inhibitors of human immunodeficiency virus type 1 (HIV-1) replication in acutely infected cells. In this study, however, we have found that some ingenol derivatives strongly enhance the replication of HIV-1 in chronically infected cells at nanomolar concentrations. One of the derivatives could activate nuclear factor kappa B (NF-kappa B), a potent inducer of HIV-1 replication, through the activation of protein kinase C (PKC). Whereas another derivative, which affected neither PKC nor NF-kappa B, significantly enhanced HIV-1 replication, suggesting that a PKC-independent mechanism may also exist in ingenol derivative-induced HIV 1 upregulation. PMID- 9856088 TI - Evaluation of recombinant A/Victoria/3/75 (H3N2) influenza neuraminidase mutants as potential broad-spectrum subunit vaccines against influenza A. AB - Current influenza vaccines require repeated administration for long-term protection. Failure to develop broad-spectrum vaccines may be attributed to the chronic presentation of hypervariable, immunodominant epitopes displayed on the viral surface that keep the immune response somewhat fixed and limited by suppression of broadly neutralizing, low-titered antibodies. To test this hypothesis, we have attempted to dampen the immunogenicity of variable epitopes and potential immunodominant domains of the A/Victoria/3/75 (H3N2) neuraminidase by site-directed mutagenesis. The results suggest that the neuraminidase structure is extremely flexible, since many substitution combinations were tolerated, and constitute proof-of-principle that the antigenicity of this protein can be modulated to a large extent. However, mice immunized with neuraminidase mutants containing multiple amino acid substitutions showed a reduced protection rate against heterologous virus in comparison with the reference groups. PMID- 9856089 TI - Antigenic and nucleotide characterization of a herpesvirus isolated from Pacific harbor seals (Phoca vitulina richardsii). AB - This report describes the antigenic and nucleotide characterization of a herpes like virus that has been isolated from the adrenal tissues of neonatal Pacific harbor seals. Infection with this virus has been previously implicated as a major cause of death of animals undergoing rehabilitation. Comparison and phylogenetic analysis of sequenced fragments of the DNA polymerase, glycoprotein B and glycoprotein D genes, and immunofluorescence assay using herpesvirus-specific monoclonal antibodies, demonstrated close similarity of the Pacific harbor seal herpesvirus to European isolates of phocid herpesvirus-1 (PHV-1) and other alpha herpesviruses affecting terrestrial carnivores. PMID- 9856090 TI - Influenza virus hemagglutinin stimulates the protein kinase C activity of human polymorphonuclear leucocytes. AB - Human polymorphonuclear leukocytes (PMNL) incubated with influenza virus, A/USSR/90/77 (H1N1) or its hemagglutinin produced a significant increase in their PKC activity when compared with untreated PMNL. The activated kinase translocated from cytosol to cellular membrane. The calcium-dependent enzyme activity was inhibited by a specific inhibitor suggesting that alpha and/or beta isoforms of PKC were involved. PMID- 9856091 TI - Inhibition of Borna disease virus replication by ribavirin in persistently infected cells. AB - Ribavirin at concentrations from 1 to 10 micrograms/ml exhibited inhibitory effects on transcription of Borna disease virus (BDV) in persistently infected cells. Our present study indicates that ribavirin is a candidate anti-BDV drug. PMID- 9856092 TI - Recombination of baculovirus DNA following lipofection of insect larvae. AB - By injection of a liposomal complex containing baculovirus DNA directly into the hemocoel of insect larvae a polyhedrosis disease was induced. After cotransfection of insect larvae with circular viral DNA and transfer vector DNA recombinant viruses were generated with a frequency of about 2%, similar to what is obtained in vitro using insect cell cultures. Based on these results an alternative strategy for the generation of recombinants can be derived for baculoviruses for which susceptible cell lines are not readily available. This strategy involves the injection of baculovirus DNA into susceptible larvae followed by in vivo cloning. PMID- 9856093 TI - Taxonomy of bacterial viruses: establishment of tailed virus genera and the order Caudovirales. PMID- 9856094 TI - Comparison of early events during infection of human and chimpanzee peripheral blood mononuclear cells with HIV-1. AB - Differences in early events during entry and integration of HIV-1 into peripheral blood mononuclear cells (PBMC) might contribute to the absence of AIDS-like disease in chimpanzees as compared to humans. To address this question, we first tested the in vitro susceptibility of human and chimpanzee PBMC for infection with the two HIV-1 isolates III B and RF. The results of these studies revealed that chimpanzee PBMC had a slightly lower capability to support the growth of HIV 1 as compared to human PBMC. This was accompanied by a delayed accumulation of proviral HIV-1 DNA in cultures of HIV-1-infected chimpanzee PBMC. However, no differences between cells of the two species were observed when very early events of HIV-1 infection were studied. Shortly (20 h) after infection chimpanzee and human cells harbored similar amounts of proviral HIV-1 DNA and PBMC of both species behaved comparably with respect to pre-integration latency (i.e. the ability to activate extrachromosomal HIV-1 intermediates in HIV-1 infected quiescent cells at various times after infection). These results strongly suggest that the absence of AIDS-like disease in chimpanzees cannot be correlated with defects in early events of the HIV-1 replicative cycle. PMID- 9856095 TI - RFLP mapping of the whole genome of ten viral isolates representative of different biological groups of potato virus Y. AB - Ten PVY isolates representative of four PVY groups (YN, YNTN, YN-W, YO), differing by their ability to induce reactions of vein necrosis on tobacco and tuber necrosis on potato, were studied in order to research the regions of the viral genome involved in these necrosis phenomena. The whole genome of these isolates was amplified in two fragments (4,063 and 5,670 nucleotides) and was subjected to a restriction fragment length polymorphism (RFLP) study. In the first 4,063 nucleotides of the PVY genome, a phenetic analysis of RFLP data resulted in a clustering of our PVY isolates into three groups: PVYN isolates (group A); PVYNTN and PVYN-W isolates (group B) and PVYO isolates (group C). In the last 5,670 nucleotides, two groups were found: PVYN and PVYNTN isolates (group D) and PVYO and PVYN-W isolates (group E). From this clustering and the necrosing properties known for these isolates, the tobacco necrosis determinants seem more likely located in the 5' than in the 3' half part of the viral RNA, whereas it would be the opposite situation for the determinants of the necrosis on potato tubers. Moreover a recombination event seemed to have occurred in the genome of the PVYN-W isolates. PMID- 9856096 TI - A thermoresistant strain of feline immunodeficiency virus (FIV) with an altered cytopathic effect and a restricted cell tropism. AB - A thermoresistant strain, designated m41, of feline immunodeficiency virus (FIV) was selected after 31 successive passages of chronically infected IRC4 cells at 41 degrees C. The wild-type virus (wt) which served as a control was cultivated the same number of times at 37 degrees C. In Crandell feline kidney cells (CrFK), the replication of m41 was similar at 37 degrees C and 41 degrees C, whereas wt multiplied only at 37 degrees C. Furthermore, m41 was more resistant than the wt strain at temperatures ranging from 37 to 56 degrees C. Syncytia formation was observed with m41 when the CrFK were incubated at 41 degrees C whereas neither m41 nor wt produced syncytia at 37 degrees C. The level of replication of wt and m41 on feline lymphoid primary cells at 37 degrees C was similar. In contrast to wt, m41 was unable to infect bone marrow macrophages. Since one or several mutations in the envelope (env) gene could be involved in changes of cell fusion properties and of cellular tropism, the nucleotide sequence of the env gene derived from wt and m41 respectively was determined. Ten mutations were found in the env gene of m41, thus leading to 9 amino acid modifications in the envelope glycoproteins. These results suggest that structural modifications of the viral envelope proteins are prerequisites for the replication of a thermoresistant FIV strain at elevated temperature and are correlated with the newly acquired viral phenotype. PMID- 9856097 TI - Dissecting the mode of action of various HIV-inhibitor classes in a stable cellular system. AB - We describe a stable and sensitive HIV evaluation system, which discriminates HIV specific membrane fusion and early transcription events and is suitable for high throughput inhibitor screening. A human lymphocytic line, constitutively producing infectious HIV-1, serves as Env-positive donor. A second indicator cell line carries a silent HIV-1 LTR lacZ reporter plasmid. A bicellular cocultivation setup allows titration and standardization of "fusion-induced gene stimulation (FIGS)" events. With few manipulations aspects of fusion and/or LTR induction can be distinguished and simultaneously assayed. Anti-Env-V3 antibodies prevent fusion and subsequent lacZ induction, and a Tat-specific inhibitor blocks only lacZ induction in a dose dependent manner without affecting membrane fusion. The LTR reporter is readily activated by Tat from HIV-1, HIV-2, or SIV and it responds to exogenous Tat protein. The reporter system is sensitive enough to detect single infection events on pre-seeded layers of indicator cells, which renders it potentially useful for direct virus quantification in patients' material. Moreover, our system allows to control and normalize DNA transfection efficiencies of HIV-derived plasmids. This aspect is particularly valuable for studies of RT- and protease-inhibitors and resistances, where p24 or supernatant reverse transcriptase, otherwise standard virus readouts, can be directly affected by inhibitors or mutations. PMID- 9856098 TI - Ultrastructural localization of nonstructural and coat proteins of 19 potyviruses using antisera to bacterially expressed proteins of plum pox potyvirus. AB - Antisera to the bacterially expressed nonstructural proteins (NSP) HC-Pro, CI, NIa, and NIb and the coat protein (CP) of plum pox potyvirus (PPV) were used for analysing the composition of virus-induced cytoplasmic and nuclear inclusions by electron microscopy. The antisera reacted with NSP and CP of PPV on immunogold labelled ultrathin sections. Antiserum to CP reacted with virions of seven out of 18 other potyviruses. CP was distributed throughout the cytoplasm of infected cells. Antisera to PPV NSP specifically reacted with virus-specific cytoplasmic and/or nuclear inclusions induced by 17 different potyviruses. NSP were furthermore localized in confined cytoplasmic areas in between complex accumulations of virus-specific inclusions. Cylindrical inclusions induced by the potyviruses were proven to consist of CI protein. Most other cytoplasmic or nuclear inclusions were shown to be composed of two or more NSP. An unexpected composition of virus-induced inclusions was observed for the crystalline nuclear inclusions of tobacco etch virus. Here, in addition to the expected presence of NIa and NIb, HC-Pro could be demonstrated. Furthermore, amorphous cytoplasmic inclusions induced by papaya ringspot virus contained the expected HC-Pro but additionally NIa, NIb and CI. Beet mosaic virus-induced nuclear inclusions ('satellite bodies') contained in their electron-dense matrix NIa, NIb, Hc-Pro and CI and in their lacunae CP in bundles of virion-like filaments. The results indicate that all cytoplasmic or nuclear inclusions of potyviruses have to be regarded as deposition sites of excessively produced viral NSP. PMID- 9856099 TI - Potyvirus aphid transmission requires helper component and homologous coat protein for maximal efficiency. AB - Aphid transmission of potyviruses depends on the presence of specific sequence domains in two virus encoded proteins, the coat protein (CP) and helper component proteinase (HC-Pro). Aphid transmissable peanut stripe virus (PStV), like most potyviruses, has an Asp-Ala-Gly (DAG) motif in the amino-terminal part of the CP. Peanut Mottle Virus (PeMoV) was determined to be highly aphid transmissible but has a unique Asp-Ala-Ala-Ala (DAAA) motif. To determine if the DAAA motif could functionally replace the DAG motif in PStV, mutations were made in a full-length cDNA clone of PStV. All of the mutations in the CP DAG motif abolished aphid transmissibility of PStV but did not affect virus infectivity. The aphid transmissibility of the PStV-DAAA mutant was partially restored by feeding aphids an artificial diet containing purified virus and PeMoV HC-Pro. The PStV-DAAA virus was poorly transmitted by aphids in vitro with HC-Pro purified from PStV or tobacco vein mottling virus (TVMV) infected plants. These experiments support the theory that specific HC-Pro/CP interactions are required for efficient aphid transmission. Based upon the sequence comparisons of 16 potyviral HC-Pro proteins several conserved motifs and striking differences have been identified. PeMoV was determined to have an Ala-Ser-Cys (ASC) HC-Pro motif instead of a highly conserved Cys-Cys-Cys (CCC) motif. We have predicted that this CCC motif could play an important role in the specific interaction between the HC-Pro and the CP DAG motif. PMID- 9856100 TI - Expression and cellular distribution of baculovirus-expressed bovine herpesvirus 1 (BHV-1) glycoprotein D (gD) sequences. AB - Glycoprotein D (gD) of bovine herpesvirus 1 (BHV-1), a homolog of herpes simplex virus gD, represents a major component of the viral envelope and is a dominant immunogen. To study the antigenic properties of the different regions of gD, we have expressed the full-length gD encoding gene and overlapping fragments spanning various regions of the gD open reading frame in a baculovirus (Autographa californica nuclear polyhedrosis virus)--insect cell (Spodoptera frugiperda, SF-9) system. Maximum levels of expression for all proteins were obtained 48 to 72 h post infection of SF-9 cells by recombinant viruses. Full length and truncated recombinant gD proteins reacted specifically with anti-gD monospecific serum as determined by immunoprecipitation and immunoblotting, indicating that the proteins retained their antigenicity. However, based on the reactivity with a panel of gD-specific monoclonal antibodies (Mabs), the full length recombinant gD lacked proper expression for two highly neutralizing linear epitopes identified by Mabs R54 and 9D6. The rest of the epitopes appeared to be preserved and antigenically unaltered. Immunofluorescence studies of recombinant baculovirus infected SF-9 cells using gD monospecific serum, revealed no direct correlation between cellular localization of the expressed proteins and their amino acid sequences. PMID- 9856101 TI - Nucleotide sequence of a resistance breaking mutant of southern bean mosaic virus. AB - SBMV-S is a resistance-breaking mutant of an Arkansas isolate of the bean strain of southern bean mosaic virus (SBMV-BARK) that is able to move systemically in Phaseolus vulgaris cvs. Pinto and Great Northern, whereas the wild-type SBMV-BARK causes local necrotic lesions and is restricted to the inoculated leaves of these hosts. Sequence analysis of the 4136 nucleotide genomes of SBMV-BARK and SBMV-S revealed seven nucleotide differences, but only four deduced amino acid changes. A single amino acid change occurred in the C-terminal region of the putative RNA dependent RNA polymerase and three differences were identified in the N-terminal portion of the virus coat protein. SBMV-BARK and SBMV-S were compared with other sobemoviruses and were found to contain a high level of nucleotide sequence identity (91.3%) to SBMV-B. Unlike SBMV-B however, SBMV-BARK and SBMV-S contained four putative overlapping open reading frames, making them more similar in genome organization to the cowpea strain, SBMV-C. The possibility exists that mutations or even errors, that resulted in mis-identification of open reading frames, occurred in previously published information on nucleotide sequence and genomic organization for SBMV-B. PMID- 9856102 TI - Chaperone protein GrpE and the GroEL/GroES complex promote the correct folding of tobacco mosaic virus coat protein for ribonucleocapsid assembly in vivo. AB - Several prokaryotic chaperone proteins were shown to promote the correct folding and in vivo assembly of tobacco mosaic virus coat protein (TMV CP) using a chimaeric RNA packaging system in control or chaperone-deficient mutant strains of Escherichia coli. Mutations in groEL or dnaK reduced the amount of both total and soluble TMV CP, and the yield of assembled TMV-like particles, several-fold. Thus both GroEL and DnaK have significant direct or indirect effects on the overall expression, stability, folding and assembly of TMV CP in vivo. In contrast, while cells carrying a mutation in grpE expressed TMV CP to a higher overall level than control E. coli, the amounts of both soluble CP and assembled TMV-like particles were below control levels, suggesting a negative effect of GrpE on overall CP accumulation, but positive role(s) in CP folding and assembly. Curiously, cells with mutations in groES and, to a lesser extent, dnaJ expressed total, soluble and assembled forms of TMV CP significantly above control values, suggesting some form of negative control by these chaperone proteins. To avoid pleiotropic effects or artefacts in chaperone-null mutants, selected chaperone proteins were also over-expressed in control E. coli cells. Overproduction of GroEL or GroES alone had little effect. However, co-overexpression of GroEL and GroES resulted in a two-fold increase in soluble TMV CP and a four-fold rise in assembled TMV-like (pseudovirus) particles in vivo. Moreover, TMV CP was shown to interact directly with GroEL in vivo. Together, these results suggest that GrpE and the GroEL/GroES chaperone complex promote the correct folding and assembly of TMV CP into ribonucleocapsids in vivo. PMID- 9856103 TI - An equine herpesvirus 1 mutant with a lacZ insertion between open reading frames 62 and 63 is replication competent and causes disease in the murine respiratory model. AB - An equine herpesvirus 1 (EHV-1) mutant was constructed by inserting a lacZ expression cassette into the intergenic region upstream of gene 62 (glycoprotein L; gL) and downstream of gene 63 (a homologue of the herpes simplex virus transcriptional activator ICP0). The recombinant lacZ62/63-EHV-1 had similar growth kinetics in cell culture to those of the parental wild type (wt) virus, with indistinguishable cytopathic effects and plaque morphology. Reverse transcriptase PCR confirmed that the lacZ insertion did not interfere with transcription of gL and immunoblot analysis indicated there was no modification to late gene expression as monitored by synthesis of EHV-1 glycoproteins C and D. The parental EHV-1 isolate HVS25A used here had almost identical nucleotide sequence to that published for isolate Ab4, in a 1200 bp region surrounding the insert, but lacked a HindIII site corresponding to Ab4 position 109,048. The lacZ62/63-EHV-1 caused respiratory disease in BALB/c mice with clinical signs, histopathology and virus titres in lungs throughout days 1-5 post infection similar to those induced by wt EHV-1. X-gal staining for beta-galactosidase expression in murine lungs clearly demonstrated EHV-1 infection in cells of the bronchiolar epithelium and pulmonary parenchyma, with a peak of infection evident at day 2 post infection, when up to 50% of bronchioles demonstrated blue-staining and thus virus-infected epithelial cells. The construction of this replication competent virus carrying a reporter gene identifies a site for insertion of foreign genes and will facilitate studies on the pathogenesis of EHV-1. PMID- 9856105 TI - Ultrastructural and physicochemical characterization of the hepatitis C virus recovered from the serum of an agammaglobulinemic patient. AB - Hepatitis C virus (HCV) morphology and physicochemical properties remain unclear because HCV usually circulates in a complexed form in association with immunoglobulins. In the present work, we were interested in the characterization of HCV particles derived from the serum of an anti-HCV negative/HCV RNA positive agammaglobulinemic patient suffering from chronic type C hepatitis. Physicochemical properties of the virus particles were determined by serum centrifugation on a 10-60% isopycnic sucrose density gradient. HCV RNA quantified by bDNA was found in a major peak at density 1.13 g/ml and in a minor peak at densities 1.05-1.07 g/ml. By electron microscopy, 45 nm large core-like particles were found at the 1.13 g/ml density while 60 nm large virus-like particles similar to other members of the Flaviviridae family were visualized at the 1.06 1.07 g/ml densities. This confirms some studies reporting the low density of HCV as compared to other members of the Flaviviridae family. PMID- 9856104 TI - Isolation of a cytopathogenic virus from a case of porcine reproductive and respiratory syndrome (PRRS) and its characterization as parainfluenza virus type 2. AB - From a lung of a fetus of a breeding sow showing PRRS-like symptoms a viral agent could be isolated. It was characterized as an enveloped, hemagglutinating RNA virus. Ultrastructural examination of purified virus revealed paramyxovirus-like pleomorphic virions of approx. 200 nm in diameter. The helical nucleocapsids were about 18 nm in diameter. The virus was found to be antigenically related to simian virus 5 (SV5) a prototype strain of parainfluenza virus type 2, but not to bovine respiratory syncytial virus, parainfluenza virus type 1, parainfluenza virus type 3, and Newcastle disease virus as determined by western blot analysis. PMID- 9856106 TI - Nucleotide sequence of the putative replicase gene of the sour cherry strain of plum pox potyvirus. AB - The complete nucleotide sequence of the NIb coding region of the sour cherry strain of plum pox potyvirus (PPV-SoC) has been determined. It consists of 1554 nucleotides and encodes a putative replicase protein of 518 amino acids. Sequence identity scores between NIb of PPV-SoC and other isolates of PPV are significantly low (c. 78%). Many of the nucleotide substitutions, however, are silent. PPV-SoC differs from isolates of PPV-D, PPV-M and PPV-E1 Amar at multiple amino acid positions that are conserved between the other isolates. The NIb sequence extends the PPV-SoC sequence presently available to 2781 nt from the 3' end (approximately 28% of the genome). PMID- 9856107 TI - Molecular characterisation of a distinct South African cassava infecting geminivirus. AB - African cassava mosaic virus (ACMV) and East African cassava mosaic virus (EACMV) are whitefly-transmitted geminiviruses (WTGs) which are widespread in cassava in Africa and cause serious yield losses. Recently, a new geminivirus affecting cassava in South Africa (SACMV) has been reported. In this work SACMV was found to have DNA-A and DNA-B components. Comparisons of amino acid sequences of the putative coat protein, and nucleotide sequences of the common region and a 687-bp DNA B fragment of SACMV with other WTGs, showed that SACMV clustered with the Old World subgroup of the Begomovirus genus of geminiviruses. Despite its bipartite nature, SACMV was most closely related to monopartite TYLCVs, but was sufficiently different to justify designating it as a distinct virus. In serological studies, SACMV grouped biologically with EACMV isolates. PMID- 9856108 TI - A reappraisal of the contribution of Friedrich Loeffler to the development of the modern concept of virus. PMID- 9856109 TI - [New HIV-1 variant discovered in Cameroon]. PMID- 9856110 TI - [Electron beam computed tomography (EBCT)--a milestone for the diagnosis of coronary heart disease?]. PMID- 9856111 TI - [Clinical value of endoscopic ultrasound-guided transesophageal fine needle puncture of mediastinal lesions]. AB - BACKGROUND AND OBJECTIVE: As the mediastinum has been a region difficult to access for biopsy, mediastinoscopy has been required in most cases. In a prospective study the value of transoesophageal endoscopic ultrasound (TEUS) guided aspiration biopsy was assessed as an alternative. PATIENTS AND METHODS: TEUS-guided fine-needle aspirations were performed between May 1995 and March 1998 in 35 patients with mediastinal space-occupying lesions. In all cases the conventional endoscopic method or percutaneous puncture-sonography had been impossible or had failed. In one patient it had been performed after a negative mediastinoscopy. RESULTS: In 34 patients (97%) the aspirated tissue cylinder could be evaluated histologically. There were no complications. Malignancy was demonstrated in 24 patients, and there were one case each of sarcoidosis, silicoanthracosis and two cases of retrosternal goitre. In four of seven patients the negative preoperative diagnosis was confirmed at operation or by follow-up. There were two false-negative results and in one patient there has been no definitive diagnosis. The accuracy of the method was thus 91.4%, the positive predictive value for malignancy 88.9% and the negative predictive value for malignancy 72.7%. Ultrasound alone was a poor predictor of malignancy in lymph node enlargement. CONCLUSION: TEUS-guided fine-needle aspiration of space occupying mediastinal lesions is an effective and low-risk method that can in selected cases shorten the diagnostic process and avoid methods that are expensive or lead to complications such as transpulmonary biopsy guided by computed tomography or mediastinoscopy. PMID- 9856112 TI - [Necrotizing hepatitis after taking herbal remedies]. AB - HISTORY AND CLINICAL FINDINGS: Two unrelated women, aged 39 and 42 years, had been admitted (at different times) to hospital because of "recurrence of an aetiologically uncertain acute hepatitis". Both patients had a history of acute hepatitis with GPT concentration of 796 and 755 U/l, respectively. Each of them had experienced recurrences of hepatitis, each of them preceded by taking herbal remedies as alternative medication, containing kava or common (or lesser) celandine, respectively. In each patient physical examination had been unremarkable. INVESTIGATIONS: Maximal values of GPT in the two patients were 422 and 350 U/l, respectively. Viral, autoimmune and metabolic causes of the hepatitis were excluded. In each of them liver biopsy revealed the picture of acute necrotizing hepatitis. DIAGNOSIS, TREATMENT AND COURSE: As it was suspected that the hepatitis was medication-induced, the intake of the mentioned herbal preparations was stopped. The liver function tests quickly became normal. CONCLUSION: In view of the rapid response to their withdrawal, a causal connection between intake of the herbal preparations and the recurrences of acute hepatitis is the most likely explanation in both cases. PMID- 9856113 TI - [Migraine: epidemiology, clinical aspects and diagnosis]. PMID- 9856114 TI - [Treatment of coronary heart disease in patients with diabetes mellitus]. PMID- 9856115 TI - [Cerebral hemorrhage during coumarin therapy]. PMID- 9856116 TI - [Proton-pump inhibitor Rabeprazole: faster acid- and symptom-control]. PMID- 9856117 TI - Orthopaedic proceedings. 1997, 1998. Abstracts. PMID- 9856118 TI - [Comparative analysis of 2 thromboplastins]. AB - BACKGROUND: This study was performed to evaluate the prothrombin time in normal healthy people (102 subjects) by means of two thromboplastins. METHODS: Dade Thromboplastin IS (rabbit brain thromboplastin) and Dade Innovin (recombinant tissue factor) were used. Derived fibrinogen, Claus fibrinogen and in vitro sensibility of these thromboplastins to known amount of heparin were also measured. RESULTS: A different behaviour of prothrombin time measurement linked to different thromboplastin sensibility connected to the age was observed. A different fibrinogen (Claus and derived) behaviour connected to the age that may help to explain thromboplastin sensibility difference with the age was also observed. Finally different sensibility of these two thromboplastins to heparin in vitro was observed. CONCLUSIONS: This result should be considered when anticoagulation is started with oral anti coagulant drug and heparin together. PMID- 9856119 TI - [Edema tester. Assessment of edema of the legs]. AB - BACKGROUND: The aim of this study was the clinical evaluation of an original device produced to evaluate edema in a semi-quantitative way, the ACI Medical, Edema Tester (ET). ET1, is a soft plastic plate characterised by two parallel conic pyramidal protrusions. One side of the protrusion is rounded while the other side has an angular edge. The two protrusions are placed on the plate in inverse decreasing height. ET2 is characterised by two lines of 7 holes placed in the plate. The ET is applied at the internal perimalleolar region with the conic pyramidal protrusions in contact with the skin. A standard sphygmomanometer cuff is applied over the area and pressure is maintained at 50 mmHg for a period between 1 an 3 minutes. The cuff is then removed. ET1: skin marks are usually just visible in normal subjects without edema and disappear in a few minutes. When edema is moderate some half of each protrusion is visible as a skin mark. In limbs with severe edema the whole length of the protrusion is clearly visible. ET2: skin marks are usually just visible in normal limbs without edema and disappear in a few minutes. In limbs with edema the number of holes visible on the skin is increased and in severe edema all holes are visible. METHODS: To evaluate semi-quantitatively the level of edema the length of the two skin marks can be measured and for the ET2 the numbers of visible holes can be counted, as they are generally proportional to the degree of edema. The ET testers were evaluated in 22 normal subjects, 19 limbs with varicose veins, 22 patients with CVI and lipodermatosclerosis, 5 patients with initial primary lymphedema and in 8 subjects with severe, chronic, lymphedema and skin alterations. RESULTS: The results showed a significant difference between normal limbs an patients. CONCLUSIONS: In conclusion, the method of evaluating edema with ET can be used to supplement the clinical and noninvasive evaluation. In general practice the presence of edema measurable with ET indicate the need for treatment. The several degrees of skin marks visibility and disappearance time may be used as a general guideline to indicate the need for different types and length of treatment. PMID- 9856120 TI - [Clinico-functional respiratory assessment of the athlete with exercise-induced asthma or bronchospasm]. AB - Asthmatic subjects can only be allowed to take part in sport activities after a careful functional evaluation of pulmonary function. For sport disciplines which require a mainly anaerobic metabolism it is enough to evaluate the respiratory function at rest, while for sport disciplines which require a mainly aerobic metabolism it is necessary to use an exercise tolerance test. The functional tests used must be able to demonstrate not only the suitability of the subjects to take up a sport, but also to recommend an appropriate sport discipline. PMID- 9856121 TI - [Diagnosis of hereditary hemochromatosis with molecular analysis of DNA in patients with anti-HCV positive liver cirrhosis. Clinical case]. AB - A case of hereditary hemochromatosis in a patient affected by anti-HCV positive liver cirrhosis is described. The difficulties for an exact diagnosis are underlined. Really, it can be particularly difficult to make a differential diagnosis between hereditary hemochromatosis and secondary hemochromatosis, if liver cirrhosis has already been found. Practically, at this stage of disease, the histological and clinical aspects of these two forms become completely interchangeable. Moreover, diagnostic difficulties increase when, at the same time, the patient presents more causes of potential liver damage. In this case report, the DNA-analysis, obtained by polymerase chain reaction amplification and enzymatic digestion, allows to make the diagnosis of hereditary hemochromatosis, because it showed the presence of two genetic mutations, considered responsible for the disease. Both the hereditary hemochromatosis and the HCV infection, had greatly contributed to the development of liver cirrhosis. In the future, DNA analysis by amplification with polymerase chain reaction, can assume relevant importance for the screening of affected patients' first grade parents too. It could permit an early diagnosis of hereditary hemochromatosis and then to start a timelier and more efficacious therapy, to prevent an irreversible histological damage. PMID- 9856122 TI - [Severe myogenic pain in an unfit subject after intensive and prolonged abdominal muscle exercise]. AB - The authors report the case of an unfit patient who, following intensive and prolonged physical exercise involving the abdominal muscles, presented a massive and diffuse subcutaneous edema (abdomen, scrotum, chest and face) together with abdominal and thoracic pain which increased in response to finger pressure. In addition, this was accompanied by a marked increase in CK, CK-MB and LDH, and TGO and TGP. Chest or heart pathologies were excluded by monitoring ECG and other clinical parameters, like heart rate and blood pressure, and by performing a chest X-ray. Muscular ultrasonography confirmed the massive subcutaneous edema and abdominal MR showed a slight edema in the suprasacral region, as well as confirming the subcutaneous edema. Hematological data gradually reduced and returned to normal after a week. Edema and pain also regressed gradually: the former finally disappeared after one week and the latter after five days. The authors conclude that clinical and laboratory findings were particularly severe because the subject was unfit and subcutaneous edema was larger than the free liquid in the abdominal cavity because the latter was absorbed by the peritoneum which acted as a dialysing membrane. PMID- 9856123 TI - [Anatomopathological and immunohistochemical findings in 6 cardiac myxomas]. AB - Heart myxoma is the most common cardiac neoplasm in adult, even if its biologic profile remains uncertain. The clinicopathologic features of 6 cardiac myxomas in patients ranging in age from 42 to 58 years are described: 5 cases were located in atria, 1 occurred in the right ventricular wall, attached by a thin pedicle to the wall next to the pulmonary artery. Grossly myxomas are generally pedunculated and average 2 to 8 cm in diameter. They appear gelatinous and polypoid, sometimes with areas of hemorrhage. Microscopic examination of specimens of myxomas removed at operation reveals the myxomatous nature of the stroma composed of abundant mucopolysaccharidic matrix, containing stellate or polyhedral cells, singly or in small clusters, and occasional blood vessels. In other cases, the matrix stains more deeply and reticulin fibers and occasional strands of collagen are evident. Immunohistochemical study reveals tumoral positivity for smooth muscle actin cells and for vimentin. In addition, endothelial cells in intramyxomatous vascular channels are positive for factor VIII and CD-34 endothelial markers. Myxomas were diagnosed in patients in whom the symptoms and signs of cardiac tumor may have been attributed to other causes. The clinical pictures produced by cardiac myxomas include non specific manifestations and mechanical interference with cardiac function. The symptoms may simulate a wide variety of other cardiac conditions (mitral valve disease, embolic phemomena, tricuspid valve disease, sudden unexpected death). A wide local excision is needed to assure that the myxoma does not recur. PMID- 9856124 TI - [Intra-articular administration of galactosaminoglucuronoglycan sulfate in moderate primary gonarthritis. A pilot study]. AB - BACKGROUND: The present study has been carried out in order to assess the clinical and functional efficacy and tolerability of galactosa minoglucuronoglycan sulphate (GGGS). The molecule has been intra-articularly administered in patients affected by moderate primary gonarthritis, according to clinical and functional parameters. METHODS: At the beginning and at the end of each period of treatment the following parameters have been evaluated: spontaneous pain, provoked pain, pain during active movement, pain during passive movement, pain at rising (7-8:00 a.m.) pain at bed time (10:00 p.m.), execution time (expressed in seconds) of up-down from a chair (five times), execution time (expressed in seconds) to walk 15 meters. Fifty patients affected by osteoarthritis of the knee were treated weekly, during one year, for two periods of six weeks each with a total amount of 12 injections. RESULTS: Treatment with Galactosaminglucuronoglycan sulphate (GGGS) seems to be effective in the therapy of osteoarthritis of the knee. The pain scores decreased and the functional levels significantly (p < 0.01) improved in a large proportion of patients and GGGS was very well tolerated. CONCLUSIONS: This study suggests that intra articular treatment is a substantially new way for GGGS administration in the therapy of osteoarthritis of the knee. PMID- 9856125 TI - [A new test for monitoring heparin therapy]. AB - A simple and easy assay for plasma heparin measurement (international units/ml) is described. The method is based upon the heparin neutralization and titration by polybrene. Heparin biological activity is measured first by activated partial thromboplastin time prolongment and then by means of polybrene titration: plasma heparin concentration was evaluated in international units/ml (IU/ml). The design and properties of this method are reported. Application in clinical practice of monitoring heparinization during hemodialysis is evaluated. PMID- 9856126 TI - [Is laparoscopic surgical practice "factual" (evidence based)? Results of a prospective regional survey]. AB - OBJECTIVE: Evidence-based medicine is a growing paradigm in health care. We conducted a prospective study to determine whether laparoscopic surgery is truly evidence-based in everyday practice. METHODS: A prospective regional survey was performed in 11 French hospitals (one university and 10 district hospitals) to ascertain how general laparoscopic surgery was conducted during the last 3 months of 1997. We also searched the electronic databases for original articles on laparoscopic procedures. The methodology of randomized trials was analyzed and procedures were classed by level of evidence. We assumed that an evidence-based procedure was which had been validated by well-designed randomized controlled or prospective trials giving homogeneous results. RESULTS: One half of the procedures performed had been evaluated by randomized controlled trials. Among the 428 laparoscopic procedures, 334 (78%) were found to be evidence-based (CI 74.1-81.9%). Twelve of the 18 indications for laparoscopy (67%) were evidence based (CI: 62.5%-71.5). There was no difference between university teaching hospitals and general district hospitals. CONCLUSION: Contrary to initial criticisms, the practice of laparoscopic surgery appears to be truly evidence based in the majority of cases. PMID- 9856127 TI - [Paragonimiasis: a rare little known disease]. AB - BACKGROUND: Paragonimiasis, caused by a lung fluke, is an parasitic disease rarely encountered in France. CASE REPORT: A 52-year-old man developed dyspnea, cough, mild fever and chest pain. Pleural effusion suggested possible pulmonary embolism or tuberculosis. Cell counts in blood and pleural effusion fluid revealed major eosinophila in this patient who had recently returned from a trip to Japan. Paragonimiasis was confirmed by ELISA. Treatment with praziquantel led to complete clinical and radiographic recovery. DISCUSSION: The clinical and radiographic features of paragonimiasis are often similar to tuberculosis with pleuropneumopathy, mild fever and dyspnea. ELISA has now replaced parasitologic diagnosis. Cure is achieved with praziquantel. PMID- 9856128 TI - [Propionibacterium acnes infectious endocarditis]. AB - BACKGROUND: Propionibacterium acnes, a Gram positive microaerophilic bacteria is considered to have a low level of virulence. Nevertheless, it can be associated with infective endocarditis. We report 2 cases and a review of the literature. CASE REPORTS: The first man developed an acute prosthetic valve infective endocarditis and died. The second case was also a prosthetic valve infective endocarditis in a patient who developed infectious spondylitis. DISCUSSION: P. acnes is an uncommon causal agent in infective endocarditis and appears to have a predilectio for prosthetic valves. A prior history of skin infection is rare. P. acnes grow quite slowly, often requiring 7 to 14 days for identification. All isolates of P. acnes from blood or valve do not necessarily mean contamination. PMID- 9856129 TI - [Spontaneous coronary dissection and migraine: coincidence or common entity]. PMID- 9856130 TI - [Cavitating pneumopathy during Salmonella typhi infection: pulmonary typhoid?]. PMID- 9856131 TI - [Aspergillus fumigatus muscle abscess revealing invasive aspergillosis in a patient with AIDS]. PMID- 9856132 TI - [Local corticosteroid therapy of lumbo-sciatica]. PMID- 9856133 TI - [Indications for extra-renal dialysis in terminal chronic renal failure. National Agency for Medical Development and Evaluation. September 1996]. PMID- 9856134 TI - [Apropos of consensus on indications for extrarenal dialysis in chronic renal failure. Interview by M. Herida]. PMID- 9856135 TI - [Death of articular chondrocytes. Mechanisms and protection]. AB - APOPTOSIS IN JOINT CARTILAGE: Apoptosis is the physiologically programmed death of cells to be distinguished from necrosis which caused non-programmed cell death. In joint cartilage, more than 50% of the chondrocytes show signs of apoptosis compared with 10% in normal cartilage. ROLE OF NITRIC OXIDE: In osteoarthritis the production of nitric oxide (NO) appears to be increased, inducing apoptosis of articular chondrocytes stimulated by interleukin 1. NO inhibitors can limit the progression of experimentally induced joint lesions in animal models. IN VITRO RESULTS: The protective effect of the chondroitin sulfates CS4 and CS6 against death of chondrocytes exposed to NO has been demonstrated in vitro. Chondrocytes for New Zealand white rabbits were cultivated in the presence of an NO donor for exposure durations from 12 to 72 hours, with or without adding CS 4&6 to the culture medium. After 3 days of culture, NO induced a significant rise in the number of apoptotic chondrocytes. In 70% of the cases, preventive treatment with 100 micrograms/ml CS 4&6 lowered the number of dead cells. This cytoprotective effect reached a mean 28% and was inversely dependent on the duration of exposure to the NO donor. PMID- 9856136 TI - [Anti-arthrosis treatments: efficacy and tolerance of chondroitin sulfates (CS 4&6)]. AB - CHONDROPROTECTIVE DRUGS: Long-acting chondroprotective drugs have a symptomatic effect. They are only effective in subjects with osteoarthritis and have no pure pain relieving effect. They act within several weeks, improve functional manifestations and have a remnant effect. CHONDROITIN SULFATES 4&6: CS 4&6 are glycosaminoglycans which participate in the matrix structure of cartilage. They are well absorbed after oral intake. They have a dose-dependent inhibitor effect in vitro on proteoglycan and collagen catabolism and have been shown to stimulate matrix synthesis. Several clinical studies have demonstrated the chondroprotective efficacy of CS 4&6 in osteoarthritis involving the hip, knee and finger joints. OSTEOARTHRITIS OF THE KNEE: A controlled randomized double blind study versus placebo was conducted in 104 patients with femorotibial osteoarthritis. The objective was to demonstrate that CS 4&6 given orally in a sequential regimen at the dose of 800 mg/d has a beneficial effect both in terms of clinical manifestations and in terms of the anatomic progression in patients with osteoarthritis of the knee. The main efficacy criteria was the Lequesne functional score. After 1 year of treatment with CS 4&6, the functional impairment was reduced by approximately 50%, a significant improvement over placebo for all clinical criteria. Tolerance was excellent or good in more than 90% of the cases. A STRUCTURE MODULATOR: This study suggests that chondroitin sulfates act as structure modulators as shown by the improvement in the interarticular space visualized on the x-rays of patients treated with CS 4&6. PMID- 9856137 TI - [Chondroitin sulfates (CS 4&6): practical applications and economic impact]. AB - PROVEN CLINICAL BENEFIT: It has been demonstrated that CS 4&6 administered at the dose of 1200 mg/d for several months is more effective than placebo and as effective as nonsteroidal anti-inflammatory drugs (NSAID) in providing pain relief and improving joint function without risk in subjects with osteoarthritis of the hip and the knee. The beneficial effect persists several weeks after the end of treatment. These advantages should thus have an overall cost-lowering effect. EVALUATION OF THE ECONOMIC IMPACT: A medico-economical study was conducted to reevaluate the beneficial effect of CS 4&6 on the quantity of NSAID prescriptions in France and to determine whether the drug was used correctly in accordance with indications at an adequate dose and treatment duration. DATABASES USED: Two databases, IMS a data bank on medical prescriptions in France, and THALES which gives information on prescriptions by 300 general practitioners, allowed a dynamic analysis of the medical files of 11,000 patients with osteoarthritis. ECONOMICAL BENEFIT: The cost of NSAID prescriptions by general practitioners was reduced by an estimated 67% in patients treated with CS 4&6. In terms of the quantity of NSAID used, the reduction in the CS 4&6 treated group was 63% and 85.3% respectively in patients treated by generalists and by specialists. The quantity of NSAIDs prescribed was reduced by two-thirds when CS 4&6 were represcribed. The cost related to CS 4&6 treatment was compensated for by the reduction in physiotherapy costs and by fewer co-prescriptions for gastroprotective drugs. This study confirmed the randomized clinical studies demonstrating that a regimen of CS 4&6 for several weeks at the dose of 1200 mg/d lowers prescriptions of NSAID which can be completely avoided in nearly half the cases. PMID- 9856138 TI - [Referred earache; an important symptom of head-and-neck cancers]. AB - Otalgia is a common symptom in general practice and represents the sensation of pain in the ear. In four patients, three women aged 63, 57 and 37 years, and a man aged 64 years, ear pain was found to be caused by laryngeal carcinoma (two patients), oropharyngeal carcinoma and nasopharyngeal carcinoma (in a Turkish patient). Referred otalgia is an earache that is caused by a nonotologic source. In many cases it is difficult to identify the underlaying disease of referred otalgia. It may be the first symptom of a head-and-neck carcinoma. In case of otalgia with normal otologic findings, the differential diagnostic process must be specifically directed to the common sensory innervation of the ear and the head-and-neck structures. By carefully taking the patient's history with special attention to epidemiological factors such as smoking and drinking habits, racial traits, a positive family history of head-and-neck neoplasms and accompanying complaints and by performing repeated ENT examination including advanced imaging techniques, long delay in diagnosing head-and-neck cancer can be prevented. PMID- 9856139 TI - [New non-steroidal anti-inflammatory drugs (NSAID's) with lesser side effects, and over-the-counter low-dose preparations]. AB - Three recent developments with respect to therapy with nonsteroidal anti inflammatory drugs (NSAIDs) are the identification of two isoenzymes of cyclooxygenase (COX-1 and COX-2), the development of NO-NSAIDs, and the availability of low-dose over-the-counter (OTC) NSAIDs. COX-1 products mainly have a physiologically regulating and protective effect, while COX-2 products result in inflammation. There is initial enthousiasm for selective COX-2 antagonists with respect to the gastrointestinal adverse reactions. However, recent animal data indicate that high doses of COX-2 antagonists are usually necessary to obtain an adequate anti-inflammatory effect, which doses also antagonize COX-1. The development of NO-NSAIDs is also promising. These drugs release NO which may protect the local microvasculature. However, there is as yet hardly any practical experience with these drugs. OTC use of low-dose NSAIDs is increasing. As far as is now known, the overall risk is low because of the lower dose used. Nevertheless, the usual contra-indications should be kept in mind, such as age over 60 years, a prior history of peptic ulcers, co-morbidity, and concurrent treatment with other NSAIDs or anticoagulants. PMID- 9856140 TI - [Non-steroidal anti-inflammatory agents (NSAID's) with lesser side effects by selective inhibition of cyclo-oxygenase-2]. AB - The efficacy and the adverse effects of non-steroid anti-inflammatory drugs (NSAIDs) are related mostly to inhibition of cyclo-oxygenase (COX), the main enzyme in prostaglandin synthesis. There are two isoenzymes: COX-1, mostly involved in the production of prostaglandins that are important for the normal function of the organism, such as protection of the stomach, vascular homeostasis and kidney function, and COX-2 which is induced by proinflammatory stimuli and is mostly present in inflammatory cells. Conceivably, the COX-1 and COX-2 functions are not separated very strictly. For instance, COX-2 may also come to expression as a reaction to physiological stimuli not connected with an inflammation. It is not yet clear how COX-2 selectivity can best be assessed. The degree of selectivity depends among other things on the test used and the time interval after which the activity is read. Because of the variable findings it is important to rank different NSAIDs on the basis of one and the same test. Use of NSAIDs that mostly inhibit COX-2 appears to result in reduction of the risk of gastric ulcer development. It is to be expected that the selective COX-2 inhibitors now being developed will make the use of NSAIDs even safer. PMID- 9856141 TI - [Roaming through methodology. IV. Causality]. AB - If a factor A is often followed by an event B, it may erroneously be concluded that one causes the other. Actually B may have been brought about by a factor C (the true cause), A being only an epiphenomenon of C. If factors other than C (for example D or E) can cause the same effect B, none of those factors is a necessary cause. Causes often are multiple: a single causal factor is often not sufficient for a certain effect to ensue, whereas that effect can result from a number of factors acting together (simultaneously or consecutively). PMID- 9856142 TI - [Writer's cramp treated with botulinum injections]. AB - OBJECTIVE: To evaluate the first clinical experience with local botulinum toxin A (BTA) injections in patients with writer's cramp. DESIGN: Descriptive. SETTING: Academic Medical Centre, Amsterdam, the Netherlands. METHOD: In May 1993-January 1996 ten patients with writer's cramp were treated with BTA (Dysport). Age of the patients varied from 28 to 68 years, the duration of complaints from 1 to 29 years. Muscles for injections were selected by observation, sometimes combined with electromyography. BTA was administered under electromyographic guidance. RESULTS: The amount of BTA administered per treatment session ranged from 15 to 400 IU. In three patients the BTA-induced weakness necessary to reach a beneficial effect on writing was unacceptable. In seven patients the response was satisfactory or good and lasted 2 to 15 months (mean: 3.5 months). CONCLUSION: The results confirm the efficacy of BTA in writer's cramp. PMID- 9856143 TI - [The importance of steroid receptors for the prognosis and hormone treatment of breast cancer patients: a retrospective study in Southeastern Netherlands]. AB - OBJECTIVE: To assess the effect of oestrogen (ER) and progesterone (PgR) receptors on the prognosis of patients with operable breast cancer and the decision to treat these patients with adjuvant tamoxifen. DESIGN: Retrospective. SETTING: Eight community hospitals in the Southeast Netherlands. METHOD: Using the registry of the Comprehensive Cancer Centre South, 2862 breast cancer patients were identified with stage I, II or IIIA tumours, treated during the period 1984-1992. RESULTS: ER and PgR status were known for 2393 (84%) and 1761 (62%) patients respectively. From 1991, over 80% of the postmenopausal, lymph node positive patients had received tamoxifen, irrespective of the steroid receptor status. Of all lymph node negative patients fewer than 3% received adjuvant systemic treatment. Among the lymph node negative patients the steroid receptor status was not a significant predictor of survival. Among the lymph node positive patients whose tumours were both ER-negative and PgR-negative, a 2.8 fold increased risk of death was found during the first four years after primary treatment. The risk of death was not increased if only the ER or only the PgR status was negative. CONCLUSION: This study shows that ER and PgR receptors are significant prognostic factors for survival in breast cancer patients with involved axillary lymph nodes. The prognostic effect appeared to be restricted to the first four years after primary treatment. Selection of patients for endocrine treatment should be based on the steroid receptor status, considering the importance of the steroid receptors for predicting the response to endocrine treatment. PMID- 9856144 TI - [Human granulocytic ehrlichiosis, a tick-borne disease]. AB - A 69-year-old male presented at a first-aid department in Connecticut (USA) with severe headache, fever and myalgia of three days' duration. Just before he became ill, an engorged tick had been noted on his back and removed. Laboratory results included a decreased white cell count and platelet count and elevated transaminases. So-called morulae consisting of microorganisms were detected in a peripheral blood smear of the day of admission. Treatment for human granulocyte ehrlichiosis with doxycycline for two weeks resulted in prompt and full recovery and after four weeks repeat laboratory tests were all within normal limits. Human granulocyte ehrlichiosis should be considered in a patient with a recent tick bite who develops fever, headache and myalgias, in the presence of leukopenia or thrombocytopenia, even in Western Europe. PMID- 9856145 TI - [Sudan, through the back door]. AB - South Sudan has five million inhabitants and has been fighting a war of independence with North Sudan since 1959. The hostilities have totally disrupted society and the country is the most inaccessible of Africa. International non government organizations co-operate in relief activities in the 'Operation Life Line'. Semi-nomadic pastoral tribes populate South Sudan. Doctors Without Borders assisted in fighting a major epidemic of kala azar in the late eighties which cost some 200,000 lives; the organization still provides medical aid in the country. PMID- 9856146 TI - [Inspection of medical education 1997]. PMID- 9856147 TI - [Hypertonic dehydration resulting from unrecognized inadequate intake of breast milk]. PMID- 9856148 TI - [Perplexing acute or chronic somatic or psychiatric symptoms: possibly due to porphyria]. AB - A woman aged 35 years had a classical picture of acute intermittent porphyria. However, the diagnosis was only made after repeated clinical investigations by internists, gastroenterologists, gynaecologists and surgeons working at a university hospital, and after an unnecessary laparotomy. Recent data indicate that acute intermittent porphyria is not a very rare disease. It may run a serious invalidating and life-threatening course. Its manifestations vary and may lead to very different clinical pictures. PMID- 9856149 TI - [The guideline 'Hearing impairment' of the Dutch College of General Practitioners; a comment from the family practitioner]. AB - The 61st guideline of the Dutch College of General Practitioners titled 'Slechthorendheid' gives clear instructions regarding anamnesis, examination, supplementary examination and management. It might be added that audiometry in general is well feasible from the age of 4 years, while the guideline gives 6 years as the lower limit. The instruction in the guideline to observe a period of watchful waiting of 3 months in adults with otitis media will give rise to problems in practice. PMID- 9856150 TI - [The guideline 'Hearing Impairment' of the Dutch College of General Practitioners; a comment from the Netherlands Society of Otolaryngology and Cervico-Facial Surgery]. AB - This guideline has been well thought out, and is a useful basis for the general practitioner to diagnose and manage hearing loss. However, it advises watchful waiting in children with otitis media, a questionable view. In evaluating simple diagnostic tools, the speech recognition is overvalued at the expense of audiometry and the tuning-fork test. Posttraumatic perforation of the eardrum, in combination with hearing loss is incorrectly presented as a self-limiting problem in the majority of cases. PMID- 9856151 TI - [Electron-beam computed tomography: preliminary experiences in the Netherlands]. AB - The difference between computer tomography (CT) and electron beam tomography (EBT) is that for CT the x-ray tube rotates in a ring round the patient, and for EBT the x-ray beam rotates itself. As a result, with EBT the speed of making images is not limited by the mechanical rotation of the tube, and 16 images can be made per second. An EBT scan of a whole thorax takes 9 seconds. Specific application areas are fast moving organs and patients who cannot remain in one position for long, e.g. children and intensive care patients. Research is being conducted into the possibilities of this non-invasive technique for the demonstration of coronary artery lesions, pulmonary embolism, pulmonary metastases and dynamic examination of the major respiratory tract. Costs of an EBT scan currently amount to Dfl. 450.--but they will probably decrease as this imaging system is developed further. The exposure to radiation is about the same as that caused by a (spiral) CT. PMID- 9856152 TI - [Summary of the guideline 'Hearing impairement' from the Dutch College of General Practitioners. The Dutch College of General Practitioners]. AB - Hardness of hearing is a growing problem; toward the year 2000 there will be 1.5 to 2 million persons with hardness of hearing in the Netherlands. The Dutch College of General Practitioners has published a guideline 'Slechthorendheid' [hardness of hearing] with instructions for the diagnosis and treatment of patients with hardness of hearing in primary care. Many patients with hardness of hearing do not visit the GP (e.g. mentally handicapped). The whispered speech test or screening audiometry plays an important part in the diagnosis. In case a hearing loss > or = 30 dB is detected consultation of or referral to an ENT specialist is an option. In most cases a plug of earwax is the cause of the hardness of hearing. PMID- 9856153 TI - [Physical diagnostic procedures: whispered speech and tuning fork test]. AB - Hearing impairment occurs in 10% of the Dutch population and may lead to major communication problems and even social isolation. A good method to detect hearing loss in a general practice is the screening audiometer. The treating physician may further use the whispered speech test and tuning fork tests which together give a good impression of the severity and the nature of the hearing loss if any. The whispered speech test is best performed in the standardized form according to the guideline 'Slechthorendheid' [hardness of hearing] of the Dutch College of General Practitioners (NHG), in which certain combinations of letters are recommended. The tuning fork tests according to Rinne and Weber indicate a difference in perception and conduction deafness, and are decisive for any hearing asymmetry. Use of the whispered speech test and of the tuning fork tests is recommended for adults and children from the age of 7. PMID- 9856154 TI - [Prevalence and relevant background characteristics of deafness and severe hearing loss in the Netherlands]. AB - OBJECTIVE: To determine the prevalence of deafness and severe hardness of hearing among the Dutch population and of some relevant background characteristics. DESIGN: Study of documents. SETTING: Dutch schools for deaf and hard-of-hearing children. METHOD: The number of deaf and severely hard-of-hearing children of 6 12 years who received education or ambulatory supervision at schools for deaf and hard-of-hearing children was counted in the period November 1996-April 1997, and a number of relevant background characteristics were collected. By means of consultation with experts, this number was corrected for children who could not be retrieved by means of the inventory at these schools. To estimate the total number of deaf and severely hard-of-hearing people in the Dutch population, this adjusted number was extrapolated to older and younger year cohorts. RESULTS: The number of deaf and hard-of-hearing children of 6-12 years, after correction for missing groups, was 0.74 (95% confidence interval (95% CI): 0.69-0.78) per 1000 persons. In 9% the age of onset of the handicap was postlingual; they more often received ambulatory supervision and more often received education at a school for hard-of-hearing children than those with a prelingual auditive handicap. Of the children 29% had a non-Western origin; they less often received ambulatory supervision, and more often received education from a school for deaf children than indigenous pupils. Among the total Dutch population, the number of deaf and hard-of-hearing persons in 1996 was estimated to be 11,400 persons (95% CI: 10,690-12,110). CONCLUSION: The prevalence of deafness and severe hardness of hearing in the Netherlands (15,490,000 inhabitants in 1996) was 11,400 persons (95% CI: 10,690-12,110). PMID- 9856155 TI - [The frequency of hearing impairment in patients with diabetes mellitus type 2]. AB - OBJECTIVE: To determine the prevalence of hardness of hearing in patients with diabetes mellitus type 2 and to compare the diabetes characteristics between diabetic patients with and without hardness of hearing. SETTING: Computerized general practitioners office in Nijverdal, the Netherlands. DESIGN: Descriptive, cross-sectional study. METHOD: In all patients aged 55 years and over, registered at the general practitioners (n = 1184), diabetes mellitus type 2 and hardness of hearing were determined. Hardness of hearing was defined as use of a hearing aid or an audiometrically determined hearing loss of at least 40db. RESULTS: A hearing aid was three to four times more prevalent in patients with diabetes mellitus type 2 (n = 86) than in subjects without diabetes of the same age (p < 0.001). Of the diabetic patients 48% had impaired hearing. Patients with hardness of hearing had a longer duration of diabetes than diabetic patients with a normal hearing (10.5 years (SD: 6.9) versus 6.9 years (6.8); p = 0.06, adjusted for age). They suffered less hypertension and had a lower level of glycated haemoglobin. CONCLUSION: Hardness of hearing is frequent in elderly patients with diabetes mellitus type 2. It may be due to diabetic neuropathy. PMID- 9856156 TI - [Addisonian crisis provoked by levothyroxine substitution therapy]. AB - A women aged 36 with a positive family anamnesis for autoimmune endocrine diseases and a history of thyroid diseases, developed major complaints of general malaise, orthostatic hypotension and loss of appetite after the start of a treatment with levothyroxin because of (sub)clinical hypothyroidism. She was found to suffer from primary adrenocortical insufficiency masked by excessive use of liquorice and a lowered metabolism, but which via the suppletion with thyroid hormone had led to an addisonian crisis. PMID- 9856157 TI - [Shoulder complaints in the family practice: a simple strategy]. PMID- 9856158 TI - [Shoulder complaints in the family practice: a simple strategy]. PMID- 9856159 TI - [Shoulder complaints in the family practice: a simple strategy]. PMID- 9856160 TI - [The medical costs of smoking]. PMID- 9856161 TI - [The relationship between serum concentration of the prostate-specific antigen and the skeletal and CT-scan in the staging of primary prostate cancer]. PMID- 9856162 TI - [Hypochondriasis]. AB - In a female patient aged 33 years and suffering from excessive concern with her physical health, hypochondriasis was diagnosed. By cognitive behavioural therapy the obsessive thoughts, controlling behaviour and avoidance behaviour were alleviated. After treatment the patient needed fewer consultations. In general practice hypochondriasis is a common problem. Patients will ask for a lot of diagnostic tests. Nothing will give them enough reassurance and there can be irritation. The general practitioner must confront them with the fact that their main problem is hypochondriasis and search for solutions. Cognitive and behavioural therapy as well as treatment with antidepressive drugs may be helpful. The earlier the diagnosis and the start of treatment, the better the prognosis. PMID- 9856163 TI - [A lack of understanding of the rulings by disciplinary committees]. AB - The fact that certain rulings of disciplinary committees evoked much criticism among colleagues is distressing in itself, but such criticism should be interpreted with due caution, as different explanations are conceivable. For instance, the published abstract of the committee ruling may have gaps with regard to facts and motivation. It is also possible that the committee has good grounds to accentuate the professional standards, which understandably may meet some resistance among the medical profession. Finally, it is theoretically possible that the disciplinary tribunal is not fully informed about relevant professional standards and current practice. It is precisely to minimize such possible communication breakdowns that disciplinary powers have largely been entrusted to persons from the medical profession. PMID- 9856164 TI - [Recent disciplinary committee rulings foster defensive medicine]. AB - Recent medicolegal rulings corrected practitioners for not applying all possible interventions in the care of acute rare conditions. The performance was judged negligent, not on the clinical information available at the time, but in the light of the later course of disease. This paper argues that quality assurance requires a system of critical self and peer assessment, superimposed on medicolegal judgement. In all instances performance should be set against (external) criteria, including guidelines. Guidelines, in particular for general practice, define optimal care, to entitle patients to what is relevant while at the same time protecting them from ill-advised interventions. The point is made that the medicolegal ruling must be based on the same criteria that govern self and peer group assessment, and not on whatever it might be possible to do (maximal criteria), to prevent quality assurance from leading to defensive practice. PMID- 9856166 TI - [Anorexia nervosa and bulimia nervosa: I. Diagnosis and treatment]. AB - Disturbed eating behaviour and disturbed body experience are important features for the differential diagnosis of eating disorders from other disorders. Eating disorders occur mainly in young females. The one-year prevalence of anorexia nervosa is 0.4% and that of bulimia nervosa 1.5% among young females. To motivate patients for treatment it is important to discuss physical problems and the high risk of severe complications with them. It is difficult to motivate patients for treatment, because anorexia nervosa patients deny their illness and bulimia nervosa patients are ashamed and hide their disturbed eating behaviour. The treatment of anorexia nervosa consists of two partly overlapping phases: normalizing the eating pattern to improve weight restoration and psychotherapeutic treatment for underlying emotional problems. Family therapy is effective for patients younger than 18 years with a short duration of illness. Cognitive behaviour therapy is the most important form of treatment for bulimia nervosa. PMID- 9856165 TI - [Excess mortality in critically ill patients after treatment with human albumin]. AB - According to the results of a systematic review of randomized clinical studies administration of human albumin to critically ill patients is associated with excess mortality, compared with withholding albumin or administration of crystalloid fluids. The study appears to be well done. Also, there are various explanatory pathophysiological mechanisms supporting the association. However, a favourable effect of albumin in certain patient groups cannot be excluded. Alternatives to albumin are available in most clinical situations, but unfortunately, they are not completely without drawbacks. The use of albumin has to be limited; it might only be abolished when a better effect of other fluids, such as synthetic solutions, is demonstrated. PMID- 9856167 TI - [Anorexia nervosa and bulimia nervosa. II. Somatic complications of undernourishment]. AB - In anorexia nervosa and bulimia nervosa, cachexia and deficient nourishment cause various physical abnormalities, especially of the endocrine and digestive systems and the heart. Disorders in the serotoninergic and dopaminergic systems contribute to development of an eating disorder, whereas an acquired deficiency of tryptophan impairs the serotoninergic system. Any problems of nutritional deficiencies, low blood sugar levels and gastrointestinal disorders disappear after normal nourishment is resumed. Hypotension and sinus bradycardia are manifestations of a physiological adjustment to a lower basal metabolism and need no treatment. Osteoporosis occurs from two years after the onset of weight loss; oestrogen supplementation may protect against this. In patients with infections, symptoms such as fever, leukocytosis and high BSE may be lacking. Hypoglycaemia incidentally leads to coma and death, and a lengthened QT interval to acute cardiac death. During restoration of the nutritional status, the intake of fluid and calories should initially be limited. During the first two weeks, the risk of cardiovascular complications is increased. PMID- 9856168 TI - [Anorexia nervosa and bulimia nervosa. IIi. Somatic complications of purging]. AB - Patients with anorexia nervosa or bulimia nervosa may try to compensate weight gain due to binge eating by purging: vomiting or use of diuretics, laxatives or diet pills. Purging may reduce the body weight through volume depletion, but has hardly any effect on the food uptake. Frequent vomiting is revealed by enlargement of the submandibular and parotid glands and rise of the serum amylase. Use of laxatives leads to electrolyte imbalances, lesions of intestinal mucosa and chronic obstipation. Physical effects of diuretics are dehydration and electrolyte imbalance; oedema may be a sign of their discontinuation. Frequent or major effects of purging are: disorders of cardiac conduction (caused by hypokalaemia), insults and cerebral dysfunction (both due to hyponatraemia). A normal serum potassium concentration does not exclude intracellular hypokalaemia; accordingly, electrocardiography is indicated. PMID- 9856169 TI - [Roaming through methodology. V. The prevention paradox: few with a high risk versus many with a moderate risk]. AB - With a good primary prevention programme a number of healthy individuals will never know that they had a narrow escape from disease and maybe even death. Large numbers of individuals participate in such a programme, while only a few of those would have fallen ill without it. Nevertheless such a population approach often will yield a larger health benefit (on a population level) than an intervention that only aims at the high risk group in the population. That has been called the prevention paradox. The choice between a population or a high risk approach has prompted lengthy debates in the Netherlands. Epidemiology offers a methodology to estimate the effects of different approaches. The health impact will be determined by the distribution of exposure in the population and the risk function of exposure. In the final decision on a prevention strategy other considerations such as cost-effectiveness of interventions, negative side effects, possibilities of identification of high risk groups and ethical issues can play a part. PMID- 9856170 TI - [Results from laparoscopically assisted vaginal hysterectomy in the Ikazia Hospital, Rotterdam, 1993-1997]. AB - OBJECTIVE: Evaluation of experience with laparoscopy-assisted vaginal hysterectomy (LAVH). DESIGN: Retrospective review. SETTING: Dept. of Gynaecology, Ikazia Hospital, Rotterdam, the Netherlands. PATIENTS AND METHODS: Of 84 patients undergoing a LAVH instead of an abdominal hysterectomy in the period 1993-1997, data on indications for surgery, hospital stay and complications were analysed from the medical records. RESULTS: Mean age was 45 years (range: 30-62). The most frequent indication was hypermenorrhoea (75; 89%). The mean duration of the operation was 87 min (range: 45-130). There was one conversion to laparotomy. Three times a major complication was seen: twice a bladder injury and once a major haemorrhage for which laparotomy was performed. Mean hospital stay was 6 days, in accordance with protocol. CONCLUSION: LAVH offers a safe technique to convert some of the abdominal hysterectomies into vaginal ones. The higher costs of disposable equipment and increased operation time must be overcome by a shorter time in hospital and shorter convalescence time. PMID- 9856171 TI - [Pellagra (deficiency of vitamin B3 or of the amino acid tryptophan): a disease still extant in the Netherlands]. AB - Pellagra was diagnosed in a 48-year-old female patient with a bullous skin disease. The skin disease with purple/red sharply demarcated spots on hands and feet had worsened after sun exposure. She was a chronic alcoholic and for the last few months she had had diarrhoea. The treatment included vitamin B3, vitamin B complex and a high-quality protein diet. Within three days her skin disease improved. Pellagra is caused by a deficiency of nicotinamide or of its precursor tryptophan. It may occur in patients with dietary deficiency diseases (e.g. chronic alcoholics), carcinoid syndrome, HIV infections and drugs: fluorouracil, isoniazid, chloramphenicol and mercaptopurine. Pellagra leads to the triad: dermatitis, diarrhoea and dementia, eventually followed by death. The skin changes are characteristic and pathognomonic. Recognition of pellagra is important; the prognosis is good after treatment. PMID- 9856172 TI - [Short-acting insulin analogs]. PMID- 9856174 TI - [Fluoride in osteoporosis: not yet validated]. AB - Over thirty years' research has failed to solve the controversy about use of fluoride in the treatment of osteoporosis. The marked increase of the mineral density of the bone in the lumbar spine is not unequivocally accompanied by a decrease of osteoporotic fractures. The therapeutic margin is narrow and there is no simple, validated method of dose adjustment. Consequently, fluoride should still be regarded as an experimental treatment, only to be used in the framework of clinical scientific research. PMID- 9856173 TI - [The immobile vocal cord]. AB - Usually dysphonia is the result of a functional disorder of the larynx. It can also result from paresis or paralysis of a hemilarynx. Four patients, men aged 57, 41, 42 and 18 years, had a neurological cause of paralysis of a hemilarynx. Processes responsible for this kind of pathology can appear at three different levels: central, nuclear and peripheral. Each of the four patients had a disorder at the peripheral level: two had a tumour, in one the vagus nerve was severed during lobectomy for squamous cell carcinoma (iatrogenic), and in the last one an upper respiratory viral infection was the probable cause. Other causes of these peripheral lesions are toxicological, traumatic or idiopathic. When dysphonia does not improve within three weeks, inspection of the larynx and palpation of the neck, including examination of the aspect and mobility of the vocal folds by an otorhinolaryngologist should be performed. If paresis of a hemilarynx is seen, an orientation examination of the cranial nerves and selective additional examination is necessary, as paresis of a hemilarynx is a symptom, not a diagnosis. PMID- 9856175 TI - [Pharmacological prevention of osteoporosis in patients on corticosteroid medication]. AB - Twenty-five per cent of the patients using corticosteroids for long periods of time develops at least one fracture. Corticosteroids lead to osteoporosis through increase of renal calcium excretion and decrease of intestinal calcium absorption, bone formation by osteoblasts and serum levels of sex hormones. In spite of guidelines according to which patients protractedly using corticosteroids should take sufficient calcium and cholecalciferol, only about one-tenth of them takes any form of medication to prevent osteoporosis. It seems advisable to prescribe additional anti-osteoporosis medication for patients using > or = 7.5 mg prednisone during at least 3 months and who have a low mineral density of bone; only of biphosphonates a preventive effect clearly has been demonstrated. For postmenopausal women, hormonal supplementation therapy may offer additional benefit, reducing the risk of cardiovascular disease. PMID- 9856176 TI - [AIDS: new developments. VI. HIV reservoirs during intensive antiretroviral treatment]. AB - Treatment of human immunodeficiency virus (HIV) infected persons with potent antiretroviral combination therapy results in a strong decline of the viral load in the blood. Whether this effect is reached in all tissues and different infected cell types is an important question. There are several potential virus reservoirs. Lymphoid tissue constitutes the largest virus compartment. With potent, often protease inhibitor containing combinations the HIV-RNA decline in lymphoid tissue runs parallel to that in the blood. The central nervous tissue is a potentially important reservoir, because of the limited penetration of several antiretrovirals, especially protease inhibitors. A few short studies with different combinations showed a decline of the amount of virus in the cerebrospinal fluid. The risk of local resistance developing is not known. There are only few studies of the effects of potent anti-HIV therapy on semen but prostate and testis tissue do not appear inaccessible to treatment. The reservoir of latently infected cells that cannot be reached by the immune system or by the viral replication inhibiting therapy, can possibly be reached with immune stimulating agents. This will cause HIV replication and cell death, while the anti-HIV therapy will prevent further replication of the produced virions. This approach is still experimental, however. PMID- 9856178 TI - [Fluoride treatment: a good choice in osteoporosis]. AB - Fluoride changes the composition of the hydroxyapatite crystal, reducing the solubility of the bone crystals. Fluoride also stimulates the production of DNA and new osteoblasts, which results in formation of new bone. The therapeutical range of fluoride is narrow. With monofluorophosphate in a dose of 76 mg twice daily positive effect on bone mass is obtained and measurement of serum fluoride concentration is unnecessary. For the treatment of osteoporosis the best result could be obtained with a combined treatment with monofluorophosphate, stimulating osteoblasts and production of new bone, and a bisphosphonate, which simultaneously inhibits osteoclasts and bone resorption. Using this combined treatment only few side effects and no lower extremity pain syndrome were observed. Patients > 65 years of age and younger patients showed similar positive effects on bone mass. PMID- 9856177 TI - [Fluoride in osteoporosis: still an experimental and controversial treatment]. AB - Fluoride has been used in the treatment for osteoporosis for more than 30 years but its use is still controversial. Fluoride stimulates the osteoblasts to form new bone tissue. However, the new bone has an abnormal texture and is less mineralized and relatively less strong. In patients with osteoporosis treated with fluoride, the bone mineral density of the skeleton increases. On the other hand, the effects of fluoride treatment on the incidence of fractures are controversial and in some studies an increased fracture incidence was observed. In addition, fluoride often causes upper gastrointestinal complaints and a lower extremity pain syndrome, which is caused by stress fractures and may last for several months. For these reasons, fluoride treatment should only take place in the framework of clinical trials. PMID- 9856180 TI - [More information on infectious diseases provided by a national information system]. AB - The Infectious Diseases Information System (ISIS) was established to improve the exchange of information about infectious diseases among those combating these diseases. Information about infectious diseases is obtained from municipal health services by means of a data processing system elaborated by the National Institute for Public Health and the Environment. After adjustment to the new Act on Combating and Tracing of Infectious Diseases it will be introduced nation-wide in 1998. Information about pathogens is obtained from medical microbiological laboratories (MMLs) through automatic storage of positive and negative results in a central data bank. After a pilot project had shown that the data from one MML, Arnhem, were complete and reliable, the MMLs of Roermond and Tilburg were also connected. For feedback of information to medical microbiologists and infectious diseases specialists, but also to, for instance, GPs, data from the whole country are presented daily on a public Internet site (http:/(/)www.isis.rivm.nl). PMID- 9856179 TI - [Annual epidemics of hepatitis A in four large cities related to holiday travel among immigrant children]. AB - OBJECTIVE: Analysis of the transmission pattern of hepatitis A in relation to ethnicity and travel behaviour in Amsterdam. Utrecht, Rotterdam and The Hague. DESIGN: Descriptive study of notified cases. SETTING: Municipal Health Services of the four major cities in the Netherlands. METHOD: Notification data of hepatitis A in Amsterdam, Utrecht, Rotterdam and The Hague were analysed over the period 1992-1995. Cases were analysed according to age (0-19 years or > 19 years), whether or not they travelled abroad in the period of six weeks before the onset of the first symptoms of disease, and endemicity of hepatitis A in the country of ethnic origin. RESULTS: The strong increase of hepatitis A after the summer holidays could be divided into several smaller epidemics starting with an epidemic among children of Moroccan and Turkish descent who had spent the summer holidays in these countries, among children of the same ethnic background who had not travelled abroad, followed by epidemics among non-travelling children and adults of mainly Dutch descent, respectively. A strong correlation was found in Amsterdam between the incidence in the former two groups and the latter two groups (Pearsons r = 0.68; p = 0.004). CONCLUSION: Children who spent the summer holidays in a hepatitis A endemic country, particularly Morocco and Turkey, appeared to be the main importers of hepatitis A in the four major cities. Active immunization of all children born in the Netherlands of Moroccan and Turkish descent is the most preferable intervention. PMID- 9856181 TI - [A wandering spleen]. AB - A mobile asymptomatic abdominal mass was accidentally found in a 81-year-old man. Further studies revealed a wandering spleen. Wandering spleen is rare, the diagnosis can easily be made with ultrasonography. The patient presented was asymptomatic, but severe complications may occur. Congenital anomalies of ligaments, splenomegaly and pregnancy may be of aetiological importance. In young patients and in case of complaints splenopexia is the treatment of choice; the patient presented was treated conservatively. PMID- 9856182 TI - [Management changes based on an ECG for patients in family practice]. PMID- 9856183 TI - [Management changes based on an ECG for patients in the family practice]. PMID- 9856184 TI - [Respiratory failure sometimes is due to a cardiac cause]. AB - In two women aged 65 and 49 years and a man aged 64 years, severe respiratory failure developed and a pulmonary disease was suspected. They also had a minor systolic murmur. At further investigation no pulmonary cause for the disease could be established. Pulmonary artery catheterization revealed increased pulmonary artery and wedge pressures and transthoracic and transoesophageal echocardiography revealed mitral valve insufficiency. Two patients had valve surgery, the third received medication. Respiratory failure is a common problem in an intensive care unit. If this condition is caused by mitral valve insufficiency the clinical picture is not always that of acute left ventricular failure with hydrostatic pulmonary oedema due to backward failure. In patients with respiratory failure due to mitral valve pathology initial diagnostic problems may be overcome by combining the findings of pulmonary artery catheterization and transthoracic and transoesophageal echocardiography. PMID- 9856185 TI - [Clinical and toxicologic aspects of the use of Ecstasy]. AB - Methylenedioxymethamphetamine (MDMA, the active compound of ecstacy (XTC) tablets) is a psychoactive amphetamine congener which in humans has a stimulatory effect and enhances feelings of openness and solidarity. MDMA is neurotoxic in animals. It depletes axonal serotonin stores, it inhibits serotonin synthesis by inhibiting tryptophan hydroxylase, and it inhibits the reuptake of serotonin into the neuron. These events lead to destruction of serotonergic axon terminals in animal brain. Selective serotonin reuptake inhibitors protect against the neurotoxic effects of MDMA. Binding of (+)[11C]McN-5652, a selective neuroligand for the serotonin transporter, is decreased in the brains of XTC-users. This indicates that XTC damages serotonergic axon terminals in human brain, also. We strongly advise against the use of XTC as the long-term clinical consequences are not known. In man, somatic life-threatening complications after XTC use include hyperthermia, hyponatraemia and liver failure. Psychiatric complications include psychosis, depression, panic disorder, and impulsive behaviour. The chronic psychosis responds poorly to therapy. PMID- 9856187 TI - [Roaming through methodology. VI. Medical perspective on the causes of death]. AB - The Central Statistics Office (CBS) annually publishes the causes of death of decreased Dutch residents. It is generally assumed that the registration of causes of death is unreliable and provides only limited additional information. It is a fact that the physicians who perform the post mortems may supply incomplete or incorrect information. Hence, individual causes of death are sometimes coded incorrectly. This necessitates specific instruction of medical students. For studies of large groups, misclassification of individual causes of death is of less importance. However, the systematics of the registration has to be taken into account. It is important that causes of death are registered uniformly facilitating (inter)national comparisons. With the aid of a causes of death register, characteristics of (groups of) the population can be related to the distribution of the causes of death; at the level of the individual patient the causes of death may afford insight into causes and effects of disease. The latter is possible by a temporary link of patient data with the individual causes of death. PMID- 9856186 TI - [Liposuction]. AB - Liposuction is a frequently applied technique in aesthetic plastic surgery, with which subcutaneous adipose tissue can be removed. In recent decades new techniques of liposuction were developed. This led to better results and fewer complications. Liposuction is indicated as symptomatic treatment of obesity and (or) lipomas. Liposuction is not without risks; patients have to be informed of practical consequences and possible complications of the technique. PMID- 9856188 TI - [Urinary antigen test as a screen for the diagnosis of beta-hemolytic streptococcal infections in newborn infants]. AB - OBJECTIVE: To differentiate between neonates with high and low risk of infections caused by group B beta-haemolytic streptococci (GBS), by using the urinary group B streptococcal antigen test. DESIGN: Retrospective. SETTING: Medical Centre Leeuwarden and Public Health Laboratory, Friesland, the Netherlands. METHODS: In a period of two years clinical, haematological and microbiological (including urinary group B streptococcal antigen detection) data were collected in newborns and their mothers who met one or more of the following criteria: a previous affected child, prolonged (> or = 12 hrs) rupture of membranes, fever in labour, unexpected preterm delivery, unexplained perinatal asphyxia. On the basis of surveillance cultures a colonization rate was made. GBS infection was 'suspected' in an unwell infant with a 'high' colonization rate; infection with GBS was 'proved' by a positive blood culture with GBS. RESULTS: 6 of 342 neonates had an infection with GBS. Risk of invasive infection increased with higher colonization rates. Sensitivity of the antigen test to detect colonization was low, sensitivity to detect neonatal infection was high (51 versus 100%). The negative predictive value of urinary antigen testing was 100%. Prolonged rupture of membranes (1.5% risk of infection) and maternal fever (5%) were the most important risk factors. CONCLUSION: In healthy neonates with risk factors but with a negative antigen detection test the risk of GBS infection is extremely low. In children with a risk factor a positive test result can indicate heavy colonization or infection. These children should be carefully observed and examined. PMID- 9856189 TI - [A marked increase in the rate of diagnosed prostate cancer in the Netherlands during 1990-1996]. AB - OBJECTIVE: To study the variation of the number of histologically detected adenocarcinomas of the prostate in the Netherlands during the period 1990-1996. DESIGN: Retrospective. SETTING: National study. METHOD: Use was made of data from the National Automated Morbid-Anatomical Record Department (PALGA), Utrecht. The following data were established for each year of the study period: the total number of histological examinations of the prostate, the number of men involved (often several histological examinations of the prostate of the same man in the same year), how often adenocarcinoma was diagnosed by these examinations and how many men were involved, as well as the age of all carcinoma patients. These data were compared with those concerning variation and incidence of cancer of the prostate obtained from the Netherlands. Cancer Registration and the Central Statistics Office. RESULTS: In 7 years a total of 205,525 histological examinations of the prostate were performed in 179,298 men; the diagnosis 'adenocarcinoma of the prostate' was made 52,964 times in 44,182 men. The number of tissue examinations with the diagnosis 'carcinoma of the prostate' increased by 63% (from 5,596 to 9,146), the number of men in whom this diagnosis was made increased by 62% (from 4,710 to 7,614). The relative frequency of prostate carcinoma in relation to all examinations of the prostate increased in 6 years (1991-1996) from 22% to 28%. In that period, the (uncorrected) incidence increased by 50%, while mortality corrected for age remained the same. CONCLUSION: Of the marked increase of the number of detected cases of carcinoma of the prostate in the study period, only a small part could be attributed to demographic changes. Since autopsies have shown that there exists a large 'stock' of subclinical carcinomas, most of the growth can probably be explained by the more intensive diagnostics with prostate-specific antigen and transrectal ultrasonography. PMID- 9856190 TI - [Frontotemporal dementia in a young woman with apparent schizophrenia]. AB - In a woman aged 35 who for the last 5 years had been suffering from conduct disorders, especially kleptomania, compulsive cleaning behaviour and affective neglect of her children, psychodiagnostic examination initially suggested a delusional disorder. In view of the gradual aggravation of bizarre behaviour, the diagnosis 'schizophrenic psychosis of the hebephrenic type' was made; supplementary examination revealed no abnormalities. Finally, in view of the ununderstood and progressive behavioural changes, the diagnosis was reconsidered; on the basis of clinical findings, SPECT examination and results of repeated CT of the brain, the diagnosis of 'frontotemporal dementia, type Pick' was made. PMID- 9856191 TI - [Hyperthyroidism induced by iodine-containing radiologic contrast media]. PMID- 9856192 TI - [Femoral neck fractures in children younger than 4 years: shorter hospital stays with the aid of traction at home]. PMID- 9856193 TI - [Early ruling out of myocardial ischemic damage by means of new biochemical markers]. PMID- 9856194 TI - [Chickenpox: varicella pneumonia in adults]. AB - Two patients, a man aged 33 years and a woman aged 30, suffered from a varicella zoster induced pneumonia. In adults a varicella zoster infection may be accompanied by a very severe pneumonia. In one patient mechanical ventilation was necessary. A chest X-ray and blood gas analysis must be made in adults suffering from a varicella zoster virus infection who have pulmonary complaints. In case of abnormalities in one of these two examinations the patient must be observed in a clinical setting. The pneumonia can be treated with aciclovir. PMID- 9856195 TI - [The laser pointer: no demonstrated danger to the eyes]. AB - If laser pointers are powerful enough (> 5 mW), they can cause ocular damage. Most laser pointers in use, however, have low power, viz. 1 mW. In the peer reviewed scientific literature worldwide not a single case of eye damage due to laser pointers is described. A review among Dutch ophthalmologists up to June 1998 revealed no cases of permanent damage caused by laser pointers. In view of the widespread use of laser pointers, the risk of retinal damage must be minimal, even with the types now banned. Laser pointers of 1 mW emitting light red or green light have sufficient visibility on projection screens. It is advisable to prohibit the sale of more powerful pointers to prevent excesses. PMID- 9856196 TI - [Overweight. I. Prevalence and trends]. AB - In the Netherlands, the prevalence of overweight and obesity defined by Body Mass Indexes (BMI, weight in kg, divided by squared height in m) of 25-30 kg/m2 and > or = 30 kg/m2, respectively, is 30-40%. As to obesity, the prevalence of 7% in males and 11% in females is intermediate between the high prevalences in the USA and modernizing and industrializing developing countries and the low prevalences in China and Japan. However, the trend over the last decade is alarming. The Netherlands is amongst the highest together with the USA, UK and Germany; mainly males have an increased risk of obesity and youngsters and subjects with low education an increased risk of overweight. PMID- 9856197 TI - [Overweight. II. Determinants of overweight and strategies for prevention]. AB - High risk groups for overweight can be defined according to features such as ethnicity, pregnancy, smoking cessation, sudden changes in lifestyle and activities, low socioeconomic status and low educational level, and a positive (family) history of diabetes mellitus and obesity. Coronary heart death is to be attributed for 15-30% to overweight (body mass index (BMI) > or = 25 kg/m2) and for 4-25% to obesity (BMI > or = 30 kg/m2). For diabetes mellitus these percentages are 64-82% and 33-75%, respectively. Population-based preventive and therapeutic measurements have to be aimed at less (fatty) food, more bodily activity, less consumption of alcohol, and better education and income. This requires policy changes at the ministries involved. PMID- 9856198 TI - [Overweight. III. Therapeutic strategies for weight reduction]. AB - Obesity is a major health hazard. It increases the risk of cardiovascular disease, diabetes, mellitus and various malignancies. Minor weight loss (5-10% of body weight) appears to reduce this risk. (Very) low calorie diets alone are not effective in the long run. Regular physical exercise and behavioral interventions improve the effect of calorie restriction to an extent that is beneficial to health. Long term drug treatment is not recommended for the treatment of obesity. A number of appetite suppressants has been withdrawn from the market because of serious side effects during long term (> 3 months) use. Also, data on the efficacy of long term (> 1 year) drug treatment of obesity are not available. However, short term use to support calorie restriction might be helpful. Bariatric surgery is indicated and reasonably safe for morbidly obese patients (body mass index > or = 40 kg/m2), particularly for those who have other risk factors for cardiovascular disease. Adjustable gastric banding appears to be an effective and safe alternative to (vertical) gastroplasty. PMID- 9856199 TI - [Lack of adherence to the guidelines on the use of fresh frozen plasma]. AB - OBJECTIVE: Evaluation of transfusion practice with regard to the guidelines for fresh frozen plasma. DESIGN: Prospective. SETTING: Academic Medical Centre, department of internal medicine, Amsterdam, the Netherlands. METHOD: During 4 weeks in April-May 1996 the indication of every request for fresh frozen plasma was established by an inquiry by telephone and compared with the indications defined by the National organization for quality assurance in hospitals (CBO). To objectivate the stated indication it was checked whether laboratory tests had been performed (prothrombin time (PTT) and activated partial thromboplastin time (APTT), and if so, what the results were. Moreover, the numbers of transfused units of red cells, platelets and fresh frozen plasma in the first 24 hours were checked. RESULTS: During the study period there were 195 requests for 844 units of plasma. 613 units were transfused in 100 patients. If the CBO guidelines were applied strictly, the proportions of plasma units inappropriately requested and inappropriately administered were 53% and 47% respectively (in 32 patients). If the CBO indications were applied less strictly, still 25% of the units were inappropriately requested and 18% inappropriately administered (in 23 patients). CONCLUSION: The guidelines were observed only moderately. Better compliance is important for medical, logistic and financial reasons. PMID- 9856200 TI - [Good results from treatment of patients with abdominal pain at the emergency department of the Amsterdam OLVG Hospital]. AB - OBJECTIVE: Evaluation of the treatment of patients with abdominal complaints at the first aid department of a city hospital, where patients were treated by emergency doctors (EMD) in combination with specialists. DESIGN: Retrospective, descriptive. SETTING: Onze Lieve Vrouwe Gasthuis, first aid department, Amsterdam, the Netherlands. PATIENTS AND METHOD: Data were collected on diagnosis, treatment and course in 3235 patients with abdominal complaints seen at the first aid department in 1997. Non-referred patients were treated by EMD who if necessary consulted a specialist. Referred patients were seen by specialists. RESULTS: Of the 2931 patients who visited the first aid department once, 1975 patients (67%) were seen after self-referral. Of this group, 1557 patients (79%) were treated by the EMD alone and could be sent to the general practitioner (GP) without consulting a specialist. These accounted for 53% of the 2931 patients with abdominal complaints. The EMD used fewer additional tests than the specialist. Of the patients, 91% could be treated conservatively, only 9% were operated. Fifty-three patients died. Analysis of the group of 304 patients who visited the first aid department more than once in the year of study, showed that in 28 cases the diagnosis was missed (in 17 cases by the EMD and in 11 by the specialist). Of this group 23 had to be operated upon. CONCLUSION: The organisation of a first aid department like that in the OLVG, with EMDs and specialists, appeared efficient; the rates of morbidity, mortality and missed diagnoses are acceptable. With relatively few additional tests, the EMD appeared to be able to differentiate between GP care and hospital care. PMID- 9856201 TI - [Rat bite fever after a bite from a tame pet rat]. AB - A 43-year-old woman presented with a generalized febrile illness, an exanthema with mixed maculopapulous and pustulous eruptions on the lower halves of the extremities, elbows, knees, palms and soles. There was also severe arthralgia and asymmetric arthritis. The diagnosis was rat bite fever. The disease became manifest eight days after she was bitten by a pet rat. Rat bite fever can easily be missed, even after adequate anamnesis and physical examination, while the differential diagnostic considerations are numerous. Our patient was cured completely after intravenous administration of penicillin G. Antimicrobial therapy was completed by an oral course of doxycycline. PMID- 9856202 TI - [Migrane treatment in the Netherlands in the early twentieth century]. AB - The treatment of migraine in the Netherlands underwent important changes in the beginning of the 20th century. Several factors played a role, including the development of analgesics (acetylsalicylic acid, phenacetin, and fampridine++) at the end of the 19th century. The introduction of phenobarbital for the treatment of epilepsy in 1912 resulted in prescription of this drug for migraine, because of the supposed similarities between both afflictions. Although ergotamine had been reported for the treatment of headache at the end of the 19th century and been recommended again during the 1920s, it appeared in the Dutch literature only at the end of the 1930s. Research abroad showed this drug to have vasoconstrictive properties in migraine, again confirming the vasogenic origin of the affliction. This is striking as vasodilating drugs had been prescribed for several years. The non-medical treatment largely remained as previously, notably the prescription of diets. The relation between migraine and anaphylaxis, based on research by the French school, was also investigated in the Netherlands and resulted in the prescription of diets to immunize against the proteins involved. PMID- 9856203 TI - [Revised guideline 'the diagnosis of dementia syndromes']. PMID- 9856204 TI - [Groin pain due to iliopectineal bursitis]. PMID- 9856205 TI - [Double vision as a symptom of a serious disorder]. AB - Three women, aged 54, 69 and 73 years, respectively, developed diplopia together with ptosis of an upper eyelid during light exercise or fatigue or continuous; in one patient the diplopia was followed by headache and vomiting. The diagnoses made were 'intracranial aneurysm', 'myasthenia gravis' and 'temporal arteritis'. Diplopia may be a symptom of a disorder timely diagnosis and treatment of which may prevent serious consequences. PMID- 9856206 TI - [The family practitioner and the postcoital test]. AB - There is still a great deal of controversy on the value of the postcoital test. Some consider it to be a valuable method for the general practitioner to evaluate the interaction in vivo between spermatozoa and cervical mucus, in spite of the poor predictive value of the test. The postcoital test is in fact only an indirect method for the detection of the production of moving sperms. As a negative postcoital test is frequently due to bad timing of the test, multiple mandatory intercourse is often necessary, which may lead to frustration and sexual dysfunction. Moreover, there are no indications that the postcoital test with its poor discriminatory power may be better in general practice than the fertility evaluations by a specialist as standardisation and reproducibility problems are equal in both settings. Therefore semen analysis appears to be a better option, for the general practitioner as well. PMID- 9856207 TI - [The total knee prosthesis: indications and complications]. AB - Severe pain, impaired movement, serious deformity and (or) instability of the knee due to osteoarthritis or another joint disease are indications for total knee replacement. Contraindications are a high surgical risk and poor function of the M. quadriceps femoris. The main problems immediately after the operation are: wound infection and prosthetic infection, palsy of the peroneal nerve and deep venous thrombosis. These occur in approximately 2% of the patients. After replacement and 3 months' rehabilitation the knee is usually free of pain and stable with a good range of movement. After 10 years, 93-98% of the operated knees are still satisfactory, and after 15 years, 85-95%. Reasons for revision are infection, mechanical loosening of components, wear of polyethylene and progressive instability. PMID- 9856208 TI - [The causes of failure for hip and knee arthroplasties]. AB - Most joint replacement prostheses in the hip or knee survive for longer than 10 years. Failure of a prosthesis may be due to infection or a fracture of bone or prosthesis, but much more often it has no clear cause, after a process of aseptic loosening. Mechanisms of aseptic loosening are: bone resorption as a reaction of bone to migrated foreign body particles, material fatigue due to repeated mechanical stress on the prosthesis and its connections with the bone, failed ingrowth due to a poor fit of the (uncemented) prosthesis in the bone bed, resorption of bone mass due to the prosthesis taking over part of the mechanical bone stresses, and wear of the material. The risk of early failure depends on the patient's bone quality and life expectation, on the surgical technique, the weight bearing on the prosthesis and the fixation, and the shape and materials of the prostheses. Selection of durable prosthetic types and techniques is being done more and more by means of stepwise introduction, (pre)clinical tests and post-marketing surveillance. PMID- 9856209 TI - [Roaming through methodology. VII. Reproducibility of measurements]. AB - Reproducibility measurements are important for a proper interpretation of medical data. Kappa is the most adequate measure for categorical variables. Kappa adjusts the observed agreement for chance agreement. The interpretation of kappa is rather difficult. The kappa value is influenced by the number of categories used for classification and the prevalence of scores of the observers. For continuous variables the Pearson correlation coefficient can be used, keeping in mind its ignoring systematic errors and its dependence on the heterogeneity of the data. Another method to assess reproducibility for continuous variables is the method of limits of agreement. This method distinguishes systematic and random errors, and quantifies the differences in the dimension of the measurements. In general, the interpretation of the various measures of agreement is helped by a visual presentation of the data in a table or figure. PMID- 9856210 TI - [The number of carotid artery operations in the Twente Medical Spectrum before and after the publication of relevant international research on the benefits of the relief of carotid stenosis]. AB - OBJECTIVE: To determine the effect of adequate scientific research on the treatment of extracranial stenosis of the internal carotid artery. DESIGN: Retrospective and comparative. SETTING: Twenty Medical Spectrum, Enschede, the Netherlands. METHOD: A comparison was made of the relevant data from 2 years of carotid artery surgery before (1989-1990; period I) and after the publication of two randomized multicentre studies (1994-1995; period II). RESULTS: The number of patients treated surgically and the number of carotid artery desobstructions had increased during period II by 339% and 319%, respectively. In period I, 25% of the patients had an asymptomatic ipsilateral stenosis of the internal carotid artery; in period II, this had decreased to 11%. In period I, 65% of the patients had a stenosis in excess of 70% of the diameter of the vessel; in period II this was 85%. The combined mortality and permanent disabling morbidity after 30 days was 6% in period I and 3% in period II. CONCLUSION: After the publication of two high-quality studies in 1991, the number of carotid artery operations increased by over 300%. The indications for the surgical treatment of stenosis were stricter rather than less strict in period II. The increase of the number of carotid desobstructions can be explained by the fact that GPs' and neurologists' referral to the vascular surgeon has changed. This change in the referring pattern may be the consequences of use of 'evidence-based' medicine. PMID- 9856211 TI - [Laparoscopic pelvic lymph node dissection for clinical staging of prostate cancer: encouraging preliminary results]. AB - OBJECTIVE: To evaluate the results of the first 72 laparoscopic pelvic lymph node dissections in patients with prostate cancer. DESIGN: Retrospective study of records. SETTING: Netherlands Cancer Institute/Antoni van Leeuwenhoek Hospital, Amsterdam, the Netherlands. METHOD: A retrospective study of records provided data on 72 patients with prostate cancer staged by laparoscopic lymph node dissection in the period 1993-1997. Per- and postoperative complications, operation time, number of removed lymph nodes, pathology result and duration of hospital stay were assessed. A comparison was made between the first series of 36 patients and the second series. RESULTS: In 9 patients the laparoscopic approach was converted to a laparotomy. This occurred six times in the first series of 36 patients and three times in the second series. The postoperative course was complicated six times in the first and four times in the second series. With increasing experience the mean operation time decreased from 140 min to 114 min in the second series (p < 0.0001). The mean number of nodes removed was equal in both series (7.5). Lymph node metastases were found in 20 patients (28%). Hospital stay was 2.9 days in the first series and 2.2 days in the second series (not significant). CONCLUSION: Laparoscopic pelvic lymph node dissection is a minimally invasive method for staging patients with prostate cancer. This staging procedure is of great benefit in patients scheduled for treatment with curative intent because of its accuracy and low morbidity. With increasing experience operation time, hospital stay and number of complications decrease. PMID- 9856212 TI - [Immediate recovery from acute renal insufficiency after abdominal decompression]. AB - A 61-year-old man received an aorto-iliac reconstruction after he was admitted because of a ruptured abdominal aortic aneurysm. Postoperatively, he developed cardiopulmonary insufficiency with anuria. After the intra-abdominal pressure had risen to 40 cmH2O (measured by a urinary bladder catheter), it was decided to perform a relaparotomy. Immediately after abdominal decompression--without correction of any other intra-abdominal pathology--the diuresis increased and several other cardiopulmonary parameters improved significantly. When a critically ill patient shows a rapid increase of the intra-abdominal pressure above a critical level an acute abdominal compartment syndrome may develop. This syndrome consists mainly of potentially fatal cardiopulmonary and renal insufficiency, for which (re)laparotomy with abdominal decompression is the only correct treatment. PMID- 9856213 TI - [Postcoital test--a useful aid for the family physician]. AB - Gynaecologists still debate the usefulness of the postcoital test. However, in general practice the duration of infertility may be shorter than in infertility outpatient clinic patients. In a primary care subfertility population (mean duration of infertility 20.7 months) it was shown that an abnormal postcoital test is associated with triple reduction in live birth prognosis. The guideline 'Subfertility' of the Dutch College of General Practitioners recommends primary investigations by the general practitioners to start when spontaneous pregnancy has not occurred after one year. Postcoital testing by the general practitioner appears to be useful. PMID- 9856214 TI - [Inhalation of sodium cromoglycate via a nebulizer: no effect on the symptoms in pre-school children with moderate asthma]. PMID- 9856215 TI - [Public health in a new coalition agreement]. PMID- 9856216 TI - [Public health in a new coalition agreement]. PMID- 9856217 TI - [The indispensable anamnesis: in-vitro fertilization in a woman under treatment for melanoma]. AB - A couple consulted the gynaecologist repeatedly because of a primary fertility disorder. When they had desired children for 8 years, no pregnancy had yet occurred. In-vitro fertilization (IVF) treatment was started. A few days after the beginning of the hormonal treatment that precedes the IVF procedure, the woman noted a swelling in the right inguinal area for which she consulted the surgeon. This swelling was found to be a metastasis of a melanoma that had been removed from her leg 6 months previously. The gynaecologist was not aware of this case history and the surgeon was not informed of the started IVF treatment. As was found later, this was in accordance with the couple's wishes. The first IVF treatment resulted in a pregnancy that ended with the birth of a healthy boy. The placenta showed melanomatous metastases and the mother died 2 months after the birth of her son. PMID- 9856218 TI - [Guidelines and costs]. AB - There is a growing interest in developing clinical guidelines which support the efficiency of medical care by weighting the potential benefits against the costs of interventions. In the recently developed Dutch guideline on reduction of serum cholesterol concentration a formal cost-effectiveness analysis is included. Based on epidemiological arguments a cost-effectiveness ratio of 40,000 Dutch guilders per life year gained was found. In comparison with other preventive health care programmes this amount was considered acceptable. In the past physicians have often taken costs into account in an implicit way when making clinical decisions. The results of the cost-effectiveness analysis vary particularly with the costs of the used statins. In the past physicians have often taken costs into account in an implicit way when making clinical decisions. However, in view of the increase in health care expenditures, it has become the responsibility of physicians to take costs into account more explicitly. Clinical guidelines with a cost-effectiveness analysis can be useful in helping physicians to provide efficient medical care. PMID- 9856220 TI - [Economic evaluation of cardiology; background and application]. AB - Effectiveness research is increasingly attracting the attention not only of policy makers but also of the medical profession. Economic evaluation may provide structured information about the cost-effect balance of an intervention as compared with that of a different intervention. Costs can be subdivided into direct and indirect medical costs and direct and indirect non-medical costs. Effects are preferably expressed in life years gained (corrected for quality). A cost-effectiveness ratio indicates the additional costs per additional life year gained (corrected for quality). Economic evaluation can be performed using local, national or international research data. It may use studies that have been completed or in which the economic evaluation is included. A (planned) subgroup analysis may help to define a group of patients for whom the cost-effectiveness ratio is most favourable. The duration of a randomized clinical trial is usually relatively brief; with the aid of a mathematical model it is possible to collect the available information and to calculate extrapolations for longer periods. PMID- 9856219 TI - [National guidelines for clinical decisions, a methodological view]. AB - Medical guidelines used to be based mostly on expertise and experience. Since the eighties they are increasingly scientifically founded. In recent years results of possible treatments have been estimated quantitatively and compared. In drawing up the consensus text 'Antithrombotic prophylaxis of vascular events in patients with manifest atherosclerotic vasculopathy' the preparatory committee, after systematic arrangement of the relevant literature, made maximal use of the results of randomized prospective clinical trials of good quality and sufficient magnitude, published in peer-reviewed journals. For most indications, the pathophysiological reasoning and the study results were in agreement. A demonstrated effect of a treatment was evaluated on the basis of its magnitude and related to the associated costs and efforts. For the consensus 'Treatment and prevention of coronary heart disease by lowering the serum cholesterol level' use was made of estimates of effects of treatment with statins versus placebo. For prevention of total mortality and non-fatal myocardial infarctions and strokes, the relative risk reduction was 30-35%. The decrease of the absolute risk depended on the initial risk. The committee was of the opinion that treatment would be useful given an absolute risk of 25% of a (subsequent) manifestation of cardiovascular disease within 10 years. This would cost Dfl. 40,000.-per year of life gained, which the committee considered acceptable. In the argumentation of guidelines there is a trend to systematic evaluation and quantitative application of the research data. Subjective assessments remain necessary, particularly the evaluation of clinical relevance of observed or assumed effects of treatments. PMID- 9856221 TI - [Melanoma in pregnancy]. AB - The incidence of pregnancy complicated by melanoma of the skin, calculated from data of the Netherlands Cancer Registry, is 1 per 10,000 pregnancies. The prognosis of a melanoma is not affected by prior or subsequent pregnancy. A melanoma diagnosed during pregnancy does appear to have a more unfavourable prognosis, due not to a less favourable clinical course, but to a delay in diagnosing melanoma during pregnancy and (or) a less favourable site. When a pregnancy is complicated by malignancy, in case of a melanoma there is a higher risk of placental metastasis compared with other malignancies. In nearly 50% of the cases reported in literature of placental metastasis there was foetal involvement. PMID- 9856222 TI - [Cholinesterase inhibitors for Alzheimer disease: preliminary recommendations for treatment. Dutch Society for Psychiatry, Section of Geriatric Psychiatry]. AB - Several cholinesterase inhibitors have recently become available for Alzheimer's disease. To reach consensus about their relevance and use in daily practice, a meeting with clinical experts was organised by the section of Geriatric Psychiatry of the Dutch Society for Psychiatry. So far, available drugs have only very modest effects on cognitive functioning and clinical impression, compared with placebo. The question whether these effects are clinically relevant cannot be answered yet. Awaiting the results of further studies, it was decided not to use cholinesterase inhibitors on a routine basis but to limit prescription of these drugs to research setting or under well-controlled conditions with regard to diagnosis and evaluation. Furthermore, prescription must be limited to mild or moderately severe dementia caused by 'probable Alzheimer's disease'. Proper evaluation in the individual patient is still an unresolved problem. Therefore, n = I protocols are to be designed. These should include the use of appropriate and standardised instruments measuring cognitive functions, behavioural functions and activities of daily life. The skills and experience required will be available in specialised and multidisciplinary units for dementia. Pharmaceutical treatment for Alzheimer's disease must be integrated with all other available forms of patient care. PMID- 9856224 TI - [The ear thermometer; not a good replacement for the rectal thermometer]. AB - OBJECTIVE: To compare the measurements of body temperature with the tympanic infrared thermometer and the digital rectal thermometer. DESIGN: Prospective, comparative. SETTING: Beatrix Hospital, Gorinchem, the Netherlands. PATIENTS AND METHODS: A total of 2057 almost simultaneous measurements of rectal and tympanic temperature were performed in 164 patients in 9 different wards. RESULTS: The mean difference between the two methods was 0.45 degree C with a standard deviation of 0.57 degree C. The tympanic temperature was lower than the rectal temperature. The differences ranged from -1.5 to 3.6 degrees C. The correlation coefficient was 0.69. If a rectal temperature > 37.8 degrees C was applied as the criterion of fever, the diagnosis was not made in 175/291 measuring moments (60%) with the tympanic thermometer. If a tympanic temperature > 37.8 degrees C was applied as the criterion of fever, the rectal thermometer failed to show fever in 16/132 measuring moments (12%). CONCLUSION: The low sensitivity of the tympanic measurement to establish fever renders the tympanic infrared thermometer unsuitable for use as a fever thermometer. PMID- 9856226 TI - [A successful treatment of drug-resistant depression with electroconvulsive therapy]. AB - A woman aged 56 had been treated at the age of 23 for a psychotic depression with vital characteristics in the context of a bipolar disorder. The treatment included electroconvulsive therapy (ECT). From the age of 47, she suffered relapses; drug treatment proved inadequate. For the last 3 years, ECT was administered, which resulted in a good condition. ECT is an effective treatment in patients with depression resistant to medication. However, in spite of continuation pharmacotherapy, the proportion of patients relapsing within 6 months after successful ECT is large. For these patients, continuation ECT may be an efficacious method. PMID- 9856223 TI - [Therapy and prevention of coronary heart diseases through lowering of the serum cholesterol levels; third consensus 'Cholesterol'. Consensus Working Group, CBO]. AB - For the second time the consensus text for lipid lowering therapy is revised. In angiographic studies it was shown that a decrease in the total cholesterol as well as the low-density lipoprotein cholesterol level results in a reduction of the progression of vascular disease. Furthermore, intervention trials demonstrated that therapy with cholesterol synthesis inhibitors reduces not only both the cardiovascular and total mortality, but also other manifestations of coronary heart disease (CHD). Hypercholesterolaemia is treated with a low-fat diet and normalisation of the weight. For individuals, this might result in a reduction of the risk for myocardial infarction or death and for the population in a decrease of the mean serum cholesterol concentration and the incidence of CHD. The indication for drug therapy is founded on the expected effectiveness to reduce the incidence of (new manifestations of) CHD, which is related to the level of the absolute risk of vascular disease. In persons without known vascular diseases this risk is calculated from the total and high-density lipoprotein cholesterol ratio, age, sex, blood pressure, diabetes mellitus, and smoking. Treatment with cholesterol synthesis inhibitors must be considered in (a) patients with familial hypercholesterolaemia, (b) all patients with a history of myocardial infarction or other symptomatic vascular disease with a total cholesterol concentration above 5.0 mmol/l and a life expectancy of at least five years; (c) persons with a combination of diabetes mellitus, hypertension, hypercholesterolaemia and high risk for development of CHD, rising from 25% per 10 years at the age of 40 years to 35-40% per 10 years at the age of 70 years, with a life expectancy of at least five years. If these guidelines are followed, the upper limit of the calculated cost-effectiveness is about Dfl. 40,000 per life year gained. The working group judges this reasonable in comparison with other therapeutic interventions in the Netherlands. PMID- 9856225 TI - [Migrants in family practice: their symptoms and diagnoses differ from the Dutch]. AB - OBJECTIVE: To determine to what extent Turks, Moroccans and Surinamese differ from the Dutch in health problems, while taking the impact of relevant background characteristics into account. DESIGN: Secondary analysis. SETTING: Dutch Institute for health care research, Utrecht, the Netherlands. PATIENTS AND METHODS: In 1987/'88, 161 general practitioners and their assistants in 103 general practices recorded all patient contacts during three months in four consecutive periods for the Dutch National Survey on Morbidity and Interventions in General Practice. Data on 1165 Turkish, 853 Moroccan, 1355 Surinamese and 1471 Dutch persons (a 1% sample) between 18 and 65 years of age were used. Differences in health problems (complaints and diagnoses) between groups were tested by means of logistic regression analysis. Sex, age, educational level, working situation, marital status, health insurance, region and urbanisation were included in the analyses as covariates, for which the data were corrected. RESULTS: The pattern of complaints and diagnoses of ethnic minorities agreed with that of the Dutch in many respects, but there were also clear differences. Surinamese differed the most. Digestive problems, acute and chronic, eye problems, acute musculoskeletal problems, especially muscle pain or fibrositis, respiratory infections and eczema occurred more often in all three minority groups. Surinamese had more diagnoses in the categories of blood and endocrine/metabolic disorders (diabetes mellitus), Moroccanshad fewer diagnoses of the circulatory system (hypertension). Turks and Surinamese had more general and social problems, while Surinamese also had more psychological problems. CONCLUSION: In some areas ethnic minorities have more health problems than Dutch patients who are comparable with them in background characteristics. This clearly suggests an 'ethnic' factor. PMID- 9856227 TI - [Pain in one leg in patients with cancer]. AB - Two patients, a woman aged 65 years and a man aged 56 years, with cancer, presented with pain in one leg as the first manifestation of metastases. The woman had tumour plexopathy of the lumbosacral plexus caused by an os sacrum metastasis of a thyroid carcinoma; she received radiotherapy but died a short time later. The man had lumbosacral epidural metastases of a colon carcinoma, compressing lumbosacral roots; with radiotherapy he survived the first year. Back pain with radiating pain is a frequent symptom in patients with cancer. Spinal epidural metastases, spinal and paraspinal metastases without epidural extension, tumour plexopathy and leptomeningeal metastases are the commonest causes. Early diagnosis (by MRI or spinal fluid examination) is important; with progressive weakness or sphincter disturbances the prognosis worsens. PMID- 9856228 TI - [Treatment of renal artery stenosis with intra-arterial stents]. AB - In some patients with hypertension or renal insufficiency, renal artery stenosis can play a causative part. If fibromuscular dysplasia is the pathology of the stenosis, treatment of the stenosis by transluminal percutaneous renal angioplasty (TPRA) results in improvement of the concomitant hypertension in 80 90% of the patients. In case of atherosclerotic lesions 50% of the patients benefit by such treatment. In renal insufficiency TPRA of atherosclerotic lesions results in improvement of renal function in only one third of the patients. As restenosis and elastic recoil are seen as the prime determinants of this lack of success in atherosclerosis, intravascular stents were developed with the aim to accomplish a permanent dilation of the arterial lumen. In recent studies stent placement resulted in cure of high blood pressure in 0-16% of the patients and in improvement in 35-70%. The percentage of improvement must be viewed with caution because of the open design of these studies. Improvement of renal function was reported in 7-36% of the patients and worsening in 8-18%. Based on these outcomes stent placement in stenosed renal arteries should not be regarded as a routine clinical treatment. PMID- 9856229 TI - [Four new drugs for glaucoma: apraclonidine, brimonidine, dorzolamide and latanoprost]. AB - Glaucoma is in most of cases initially treated with drugs, viz. beta-blocking agents, miotics, sympathicomimetics and carbonic anhydrase inhibitors. The therapy of first choice is a beta-blocking agent, but in approximately 50% of the patients treated the effect becomes inadequate with time and combination therapy is necessary. Recently, four new antiglaucomatous agents have become available: apraclonidine, brimonidine, dorzolamide and latanoprost. Apraclonidine, an alpha 2-adrenergic agonist, is indicated for brief episodes of postlaser rise of the intraocular pressure. Longer treatment may lead to tolerance. Brimonidine, another alpha 2-adrenergic agonist, is indicated for long-term treatment of glaucoma; tolerance does not often occur. Dorzolamide is a local carbonic anhydrase inhibitor which lacks the systemic side effects seen after oral administration of carbonic anhydrase inhibitors. Latanoprost, a prostaglandin F2 alpha-derivative induces an effective decrease of the intraocular pressure if administered as monotherapy and has a good efficacy when combined with other drugs lowering the intraocular pressure. The new antiglaucomatous agents are a welcome addition to the pharmacotherapy, since in many cases they make it possible to postpone or avoid surgery. PMID- 9856230 TI - [Roaming through the methodology. VIII. Pilot studies: sense and nonsense]. AB - A pilot study is often performed before the start of a large study. Its aim is improving the methodological quality and evaluating the feasibility. The results of a pilot study are also often used to gain an impression of the efficacy of an intervention. For this purpose a pilot is absolutely unsuited. However, the estimate of the effect of an intervention in small studies such as pilot studies is determined to a large extent by chance. When conducting small studies the chance of publication bias is large. Small studies of adequate methodological quality should be published, whether the results are positive or negative, significant or non-significant, because combination in a later cumulative meta analysis may lead to sufficient power to assess the efficacy of an experimental intervention. PMID- 9856231 TI - [Continued driving habits of patients diagnosed with excessive daytime sleepiness]. AB - OBJECTIVE: Description of driving behaviour of patients with excessive daytime sleepiness (EDS) and the influence on their driving of diagnosis, therapy, legal guidelines and advices. DESIGN: Inventory based on questionnaires. SETTING: Centre for Sleep and Wake Disorders, The Hague, the Netherlands. METHODS: All patients with EDS who were in a stable condition for at least 6 months after therapy were sent a questionnaire (n = 138). The questions concerned actual driving behaviour, EDS and the effects of diagnosis, therapy and advice given by the Centre. RESULTS: The response was 67% (n = 92). Eighteen patients had never driven a car. Eighteen of the other 74 patients quit driving or drove less. Fifty six patients did not change their behaviour or drove even more. There was little effect on driving behaviour of diagnosis, therapy and advice to stop driving. CONCLUSION: Patients with EDS often participate as drivers in modern traffic. Legal guidelines and advice given by physicians have only little influence on this behaviour. As general measures seem to be ineffective, approaches tailored to the individual patient may lead to more acceptable results. PMID- 9856232 TI - [A daughter with Turner's syndrome: the impact on parents]. AB - OBJECTIVE: To inventory the problems encountered by parents of girls with Turner's syndrome. DESIGN: Descriptive. SETTING: University Hospital Rotterdam Sophia Children's Hospital, department of Child and Adolescent Psychiatry, Rotterdam, the Netherlands. METHOD: In 1995, structured interviews were held with 65 parents/pairs of parents (total 119 parents) of 66 girls with Turner's syndrome (36 aged 6-11 years and 30 aged 12-18 years) who participated in studies of the effect of growth hormone treatment. The questions concerned the parents' emotional reactions immediately after hearing the diagnosis and at the time of the interview, and the inner experience of their child's development. RESULTS: Immediately after the diagnosis, most parents felt sadness (95%) and shock (82%), approximately half were angry (61%) and ashamed (56%) and a minority felt guilt (29%). At the time of the interview (in 100 instances (84%) at least 5 years later), one-third of the parents still felt sadness, shock or anger and half felt ashamed. During the first few years of life, half the girls had been hospitalized at least three times, and feeding and sleeping also gave problems. Subsequently, the parents had problems with retarded motor development (67%), the acquisition of language (50%) and the restricted social skills of the child (67% were ragged a lot). Nearly all parents (90%) found their daughter's infertility difficult to cope with. They expected fewer opportunities for their daughter to find a job (38%) and (or) a partner (54%). CONCLUSION: Parents of girls with Turner's syndrome frequently find it difficult to cope emotionally with the fact that their child has this disorder, and with the problems regarding their daughter's linguistic and motor development and subnormal social skills. PMID- 9856233 TI - [Factitious insulinoma]. AB - A 32-year-old woman was admitted with signs of recurrent hypoglycaemia. Within 72 hours hypoglycaemia was successfully provoked by prolonged fasting. Also, blood samples demonstrated high levels of serum insulin and C-peptide and the insulin glucose ratio was abnormally high. An insulinoma was strongly suspected. However, extensive imaging displayed no tumour in the pancreas. The patient also had extensive psychological and social problems. The psychiatrist suggested a factitious disorder. High serum concentrations of insulin and C-peptide in combination with the psychiatric disorder led to the suspicion of abuse of sulfonylurea derivatives by the patient. This was confirmed by toxicological screening. A patient with unexplained hypoglycaemia, especially if an insulinoma cannot be detected, should be suspected of abusing sulfonylurea derivatives. PMID- 9856234 TI - [How long should prophylactic antipsychotic therapy last after the initial schizophrenic psychosis?]. PMID- 9856235 TI - [Increased risk of death in critically ill patients after treatment with human albumin?]. PMID- 9856236 TI - [Increased risk of death in critically ill patients after treatment with human albumin?]. PMID- 9856237 TI - [Increased hearing loss in patients with diabetes mellitus type 2]. PMID- 9856238 TI - [Economic evaluation in cardiology: background and application]. PMID- 9856239 TI - [Infection with Mycobacterium genavense in 2 HIV-seropositive patients in Amsterdam]. PMID- 9856240 TI - [Fracture of the femur neck in children younger than 4 years: shorter hospital stays with the aid of traction at home]. PMID- 9856242 TI - Spinal drop metastases in recurrent glioblastoma multiforme. AB - Multifocal dissemination of glioblastomas is very rare but is increasing as patients live longer. Between April 1994 and December 1997 one hundred and fifty one patients with a histologically proven glioblastoma multiforme were operated on in the Neurosurgical Department of the University of Kiel, Germany. Recurrent tumours of these patients were removed in 36 patients. Two patients developed multifocal spread of glioblastoma multiforme including spinal drop metastases. Both patients died 10 and 7 months after the primary operation. On histological examination both tumours showed wide perivascular tumour-cell cuffings in the surrounding brain tissue, so that this perivascular growth might be another explanation for the dissemination in these glioblastomas. PMID- 9856241 TI - Technical principles for protoporphyrin-IX-fluorescence guided microsurgical resection of malignant glioma tissue. AB - Malignant gliomas accumulate fluorescing protoporphyrin IX intracellularly after exposure to 5-aminolevulinic acid, a metabolic precursor of haem. This phenomenon has been exploited for intraoperative identification of residual tumour to enable greater completeness of tumour removal. The present report describes the necessary modifications to the operating microscope to enable microsurgical, fluorescence-guided tumour removal. The system consists of a xenon light source coupled to the microscope, which can be switched from normal white light to violet-blue excitation light (375-440 nm). A longpass filter is introduced into the observer light path to enable observation of tumour fluorescence. Transmission characteristics of excitation and observation filters are chosen to transmit part of the remitted excitation light. Thereby the observer retains an impression of tissue detail, next to tumour porphyrin fluorescence. An integrating three chip CCD camera optimized for red light detection enables documentation of fluorescence findings. The present modifications allow uncomplicated and rapid recognition of red tumour fluorescence and its borders to normal tissue, without interrupting the course of the operation. Tissue detail is great enough to enable tumour resection under violet-blue excitation light during parts of the operation. The system appears to constitute a useful tool for optimizing removal of malignant gliomas on a routine basis. PMID- 9856243 TI - Posterior fossa meningiomas: surgical experience in 52 cases. AB - BACKGROUND: Early reports of the surgical management of posterior cranial fossa meningiomas (PCFM) yielded poor results with high rates of mortality and morbidity. With the advent of modern neuroimaging and microsurgical techniques the results of surgery have improved markedly, but despite these advances removal of these lesions remains a challenge. METHODS: The results of the surgical treatment of PCF meningiomas were examined with the aim to identify particular features associated with increased mortality and morbidity. RESULTS: Of 713 patients with meningioma, 52 patients were identified with PCFM. Total macroscopic excision was achieved in 44 patients (84%). Postoperative complications occurred in 28 patients (54%) with permanent sequelae in 18 (35%). There were no mortalities in the immediate postoperative period. Follow-up ranged from 14 to 174 months (mean 42), tumour has recurred in 11 patients (21%) with a long-term mortality of 11%. At their latest follow-up 41 (79%) of patients achieved Glasgow outcome scores of 4 or 5. CONCLUSIONS: Total excision of tumour should remain the goal of treatment in patients with PCFM. Despite the recent advances in preoperative planning and surgical techniques, the morbidity associated with surgery remains significant. Notwithstanding, the majority of patients achieve a good outcome with surgical treatment. PMID- 9856244 TI - Meningioma surgery in the elderly: a case-control study assessing morbidity and mortality. AB - GOAL: To assess the morbidity and mortality of meningioma surgery in patients over age 65 and compare our results with matched controls and with the present literature on meningioma surgery in the elderly. METHODS: An evaluation of 114 patients undergoing meningioma resection divided into two groups: 57 patients aged 65-87, and a control group of 57 patients aged 25-64 matched by ASA status and tumor site. Operative complications, 30-day mortality, and pre- and postoperative neurologic status were assessed with follow up one to three months. RESULTS: Complication rates in the two groups were similar and were low. Four out of fifty-seven elderly patients (7.0%) had a surgical complication compared with five younger patients (8.8%) Excluding asymptomatic DVT detected by screening, three elderly patients had medical complications (5.2%) compared with two controls (3.5%). The vast majority of patients-93% of the elderly group and 89.4% in the controls--experienced either improvement or no change in neurologic status at followup one to three months after surgery. There was one death among elderly patients within thirty days for a mortality rate of 1.8% compared with no mortality in the case control group. The elderly patient who died was an 80-year old man who died of pneumonia in a rehabilitation hospital three weeks after surgery. Our study group of 57 patients was compared by tumor site and preoperative general health status to other series in the literature. In a review from the literature of 417 patients over age 65 who underwent meningioma surgery, the average 30-day mortality was 16%. The complication rate averaged 39%. DISCUSSION: Our lower morbidity and mortality rates after meningioma surgery in the elderly may be a result of patient selection, surgical technique, or excellent pre- and post-operative care by the resident staff, anesthesiologists, and nurses. It suggests, however, that elderly patients can have meningioma surgery safely. PMID- 9856245 TI - Outcome after aneurysmal subarachnoid haemorrhage: the use of a graphical model in the assessment of risk factors. AB - In 102 consecutive prospectively identified patients with subarachnoid haemorrhage (SAH) we have analysed the severity of the initial haemorrhage and the direct and indirect effects of adverse factors on outcome. The data we recorded included delay in diagnosis, risk groups, Doppler measurements, angiographic findings, surgical events and outcome at 1 year. By using a temporal graphical chain model, the associations between all variables and possible causal pathways were statistically determined. The severity of the initial haemorrhage, as determined by means of a clinical assessment and CT scanning, allowed low-, medium- and high-risk patient groups and a statistically predictable outcome to be identified. The overall management mortality was 13.7% at 1 year; 70.6% had a favourable outcome and 15.7% were severely disabled. Outcome was directly associated with risk group (p = 0.0038) and rebleeding (p = 0.0000). Delayed diagnosis led to a poorer outcome (p = 0.014)--an indirect association probably due to rebleeding. Adverse surgical events led to a significantly poorer outcome in high-risk patients. No significant relationship was found either between age and risk group (p = 0.7784) or between age and outcome (p = 0.6418). Preoperative clinical (WFNS) grade was unreliable in predicting outcome. It is the particular risk group, determined by the initial SAH, that indicates the individual patient's outcome. Management strategies can reduce preventable adverse events such diagnostic delay and rebleeding. Future studies should stratify patients according to risk group, delay in diagnosis and rebleeding in order to enable a clearer comparison to be made of treatment methods. PMID- 9856246 TI - Transcranial cerebral oximetry related to transcranial Doppler after aneurysmal subarachnoid haemorrhage. AB - Noninvasive methods for detecting cerebral artery vasospasm, still a serious complication following aneurysmal subarachnoid haemorrhage, are of vital interest. Up-to-date transcranial Doppler ultrasound (TCD) has proved to be sensitive in detecting vasospasm in the middle cerebral artery, but has less accuracy for other cerebral arteries. Transcranial cerebral oximetry (TCCO) is a new non-invasive technique which may increase the reliability for detecting cerebral ischaemia. The purpose of the present study was to evaluate a putative correlation between TCCO and TCD. We examined the two hemispheres in 14 patients with the aim of evaluating a proposed correlation between TCD and TCCO. Analysis of all absolute values (maximum TCD mFV and minimum TCCO saturation, respectively) in all series indicate a correlation between TCCO and TCD, p < 0.01, r = -0.62. All patients with TCD mean flow velocity > 120 cm/s also presented TCCO saturation < 60%. Conversely, all patients with normal TCCO saturation (> or = 63%) presented normal or moderately increased TCD velocities. In clinical neurosurgical practice it is of great interest if a true correlation between TCD and TCCO exists. The present results support the assumption that TCCO may enhance the reliability for detecting cerebral ischaemia after aneurysmal subarachnoid haemorrhage. PMID- 9856247 TI - An additional variant of the persistent primitive trigeminal artery: accessory meningeal artery--antero-superior cerebellar artery anastomosis associated with moyamoya disease. AB - Persistent embryological carotid-basilar anastomoses are rare. There has been no report on persistent anastomosis between the accessory meningeal artery and the antero-superior cerebellar artery. We describe a 3-year-old boy with moyamoya disease manifesting such a congenital vascular anomaly, namely, a large anastomosis between the accessory meningeal artery and the antero-superior cerebellar artery associated with marked hypoplasia of the basilar artery. This persistent vascular anomaly is considered embryologically as an additional variant of the persistent primitive trigeminal artery, i.e., a "stapedo-trigemino cerebellar" variant. PMID- 9856248 TI - Intellectual functions and regional cerebral haemodynamics after extensive omental transplantation spread over both frontal lobes in childhood moyamoya disease. AB - Most accepted operations for childhood moyamoya disease have attempted to increase cerebral blood flow (CBF) in the ischaemic cortical areas around the central fissure. Developed ischaemic brain damage in the prefrontal area may lead to poor intellectual outcome and restrict patients' daily lives. Thus, extensive cerebral revascularization in both the ischaemic anterior and middle cerebral artery territories is mandatory. We describe the long-term follow-up results for intellectual outcome and performance status and make an evaluation of regional cerebral haemodynamics after extensive omental transplantation spread over both frontal lobes performed as the initial management. In the past 10 years, 10 moyamoya patients less than 12 years of age consecutively underwent omental transplantation. The omental flap was spread over not only the symptomatic hemisphere but also the contralateral frontal lobe after a large craniotomy. Superficial temporal artery to middle cerebral artery (STA-MCA) anastomosis was accomplished simultaneously. On the contralateral hemisphere, STA-MCA anastomosis combined with encephalomyosynangiosis was subsequently performed. The clinical observation period averaged 6.7 years (ranged 1.9 to 9.2 years). Apart from 2 patients in whom severe mental retardation had been disclosed pre-operatively, full-scale intelligence quotient scores have been maintained at over 90, that is, within the normal intellectual range. With respect to quality of life (QOL), these 8 patients have been leading normal daily lives since the operation. The focal decrease in CBF observed in the frontal lobe pre-operatively in 7 cases had disappeared after surgical treatment. In these patients, serial post-operative MR angiography revealed developed omental vessels and STAs. Deterioration of intellectual functions and QOL as well as cerebral ischaemic events in paediatric moyamoya patients can be prevented by extensive omental transplantation spread over both frontal lobes combined with STA-MCA anastomosis. PMID- 9856249 TI - Closed reduction of bilateral locked facets of the cervical spine under general anaesthesia. AB - BACKGROUND: Bilateral facet interlocking of the cervical spine is a relatively uncommon type of cervical spinal injury. It is frequently associated with devastating neurological symptoms and signs. Early reduction of the locked facets is thought to be critical in preventing progressive secondary spinal cord injury. Whereas skull tong traction remains our primary option for closed reduction of bilateral locked facets of the cervical spine, it is not always successful, even with heavy traction weights. Other more aggressive measures may occasionally be required. The authors report their experience in reducing bilateral locked facets of the cervical spine by manual closed reduction. METHODS: This small series consists of six cases of cervical spinal injury with bilateral locked facets in which manual closed reduction under general anaethesia and muscle relaxation was used. Three of them presented with complete quadriplegia (Frankel class A). One case presented with incomplete but severe neurological deficits (Frankel class B). After unsuccessful closed reduction with skull traction, these patients were treated by manual closed reduction under general anaesthesia and muscle relaxation, followed by anterior discectomy, interbody fusion and stabilization. RESULTS: All cases made neurological improvement after the procedures. Even in cases with initial severe neurological deficits, the recovery was remarkable. The recovery was dramatic in two cases. Case 1 improved from Frankel class B to E; and Case 5 from Frankel class A to D. No case deteriorated neurologically after the procedures. Pneumonia occurred in Case 3; and stress ulcer accompanied by haemorrhage was noted in Case 4. None of these complications was directly related to the procedures. CONCLUSION: The potential for improvement of neurological function following early and successful reduction and fixation of the dislocated spine is emphasized. With meticulous techniques, manual closed reduction may be an effective alternative to skull tong traction when the latter fails. PMID- 9856250 TI - Obligatory roles of protein kinase C and nitric oxide in the regulation of cerebral vascular tone: an implication of a pathogenesis of vasospasm after subarachnoid haemorrhage. AB - We previously showed that a canine basilar artery manifested tonic and potent, protein kinase C (PKC)-dependent contractions when nitric oxide (NO) was inhibited. We also reported a linear correlation between chronological changes in the angiographic severity of vasospasm, enhanced PKC, and attenuated guanosine, 13',15'-cyclic monophosphate (cGMP) activity in a canine subarachnoid haemorrhage model. The activity of cGMP is an indicator of NO-function. Based on this evidence, we have hypothesized that PKC and NO regulate cerebral vascular tone. We particularly focused on the role of NO in a negative feedback mechanism on PKC activity in the maintenance of vascular tone. To further confirm our hypothesis, we investigated the effect of PKC down-regulation on the tonic vascular contraction induced by NO-inhibition. Canine basilar artery was used in the experiment. Significant down-regulation of PKC activity in vascular smooth muscle cells was obtained by incubation with 10(-5) mole/L of phorbol 12-myristate 13 acetate (PMA) for 24 hours. The tonic and potent contraction induced by NO inhibition was completely suppressed in the PKC down-regulated artery, even though the artery manifested a significant contraction in high-K+ solutions. These results indicate an obligatory role of PKC activity in tonic contraction when NO is inhibited, and support our previous data. Nitric oxide induces vascular relaxation by inhibiting PKC activity. Subarachnoid haemorrhage impairs this inhibition, resulting in PKC-dependent vascular contraction, such as vasospasm. PMID- 9856251 TI - Sinusoidal intrathecal infusion for assessment of CSF dynamics in kaolin-induced hydrocephalus. AB - OBJECTIVE: To evaluate whether changes of CSF outflow resistance and compliance in hydrocephalus can be assessed by an intrathecal infusion which is performed at a sinusoidal varying rate. METHODS: Hydrocephalus was produced in 10 Sprague Dawley rats by instillation of 0.0375 g of kaolin in 0.9% saline into the cisterna magna. Measurements were performed 4 weeks later: With each animal both, three successive constant rate infusions (0-0.02 ml/min) and a sinusoidal infusion (0-0.02 ml/min, frequency 0.006 Hz) were performed. 6 normal animals served as control. The pressure recordings of both infusion techniques were used for the assessment of the CSF outflow resistance. The time constant and the pressure volume index were calculated only from the sinusoidal input testing. RESULTS: The sinusoidal test as well as the constant rate infusion both demonstrated a severe impairment of CSF absorption. By the sinusoidal input, a decreased compliance was confirmed additionally. Thus, the sinusoidal infusion test demonstrated a high resistance and low compliance hydrocephalus in the kaolin-treated group. A simple graphical procedure is presented which allows an easy assessment of CSF dynamics by the sinusoidal infusion test. PMID- 9856252 TI - Experimental arterial lesions after narrow-beam gamma irradiation used in stereotactic radiosurgery. AB - An experimental study on rats showing the arterial lesions caused by the beams of gamma irradiation used in stereotactic radiosurgery is presented. The common carotid artery of the rat was irradiated with a single narrow-beam of gamma radiation with a maximum dose of 30 and 60 Gy. The results were evaluated at 4, 12 and 24 weeks after irradiation. Several focal changes in the arterial walls were observed in all periods of study with a predominance of hyperplastic (proliferative) alterations at doses of 30 Gy, and hyaline degeneration at doses of 60 Gy. Total occlusion was not observed in any case. In conclusion, vascular hyperplasia is more prominent with the lower doses evaluated. PMID- 9856253 TI - Neoplasm of endolymphatic sac origin: clinical, radiological and pathological features. AB - This paper reports on a 55-year-old female who had undergone middle ear surgery 12 years previously and was admitted with a 6-months history of unilateral hearing loss and facial weakness. MRI and CT demonstrated a space-occupying lesion arising from the temporal bone and extending into the posterior fossa. Treatment consisted in complete tumour removal. Temporal and mastoid bone destruction associated with typical histological features led to the diagnosis of neoplasm of endolymphatic sac origin. Clinical, histological, radiological and intra-operative features of these rare tumours are described and discussed. The pertinent literature is reviewed. PMID- 9856254 TI - Enterocutaneous fistula: a complication of posterior iliac bone graft harvesting not previously described. AB - A case of enterocutaneous fistula at the donor site is presented. A patient underwent posterior C5-C6 wire fusion with autologous bone graft taken from the posterior superior iliac crest for degenerative C5-C6 spondylolisthesis. The tip of the osteotome slipped anteriorly during the procurement but neither neurovascular nor peritoneal injury were observed. Spillage of formed faecal material was observed from the donor site on the 12th postoperative day. Fistulogram showed an enterocutaneous fistula to descending colon. The fistula closed spontaneously in a week. Enterocutaneous fistula after bone harvesting has never been reported in the literature as far as we know. Congenital malformations and acquired causes may create a vulnerable peritoneal area. Small bowel or descending colon may show a close relationship or even an adhesion to peritoneum in the presence of local peritonitis. A small lesion in this area may play a role in the occurrence of a fistula, and a low flow pseudofistulous tract may appear. The thermal injury and possible ischaemic necrosis due to cauterisation may be predisposing factors in the aetiology of this kind of fistula. The lack of foreign material and spontaneous closure were against an infectious origin. The aims of this report are to present and analyse the reasons of this complication, not previously described. PMID- 9856255 TI - Pyogenic osteomyelitis of an invaginated odontoid process with rapid deterioration of high cervical myelopathy: a case report. PMID- 9856256 TI - Systemic antibiotic therapy on multiple intracerebral abscesses of unknown origin. AB - A 68-year old patient presented with increasing right-sided weakness, dysarthria, pyrexia and a deteriorating general condition. CT and MRI showed about 20 round hyperdense lesions with peripheral enhancement of contrast material up to 1.5 cm in diameter. Antibiotic triple-therapy using ceftriaxone, gentamycin and metronidazole for 39 days and followed by antibiotic double-therapy using ceftriaxone and metronidazole for a further 22 days resulted in a radiologically proven reduction of the abscesses to very small remnants which disappeared completely during the follow-up period of two years. Simultaneously the general condition of the patient improved significantly. Repeated attempts at isolation of bacteria, fungi, protozoa and parasites from a subdural empyema failed. The histological examination of an abscess, which was entirely removed on occipital craniotomy, showed a structure resembling actinomycosis. PMID- 9856257 TI - Bilateral posterior fossa subdural haematomas secondary to anticoagulant therapy. PMID- 9856258 TI - The anatomical aspects of a surgical approach through the floor of the fourth ventricle. PMID- 9856259 TI - [Physiopathology of contact eczema]. PMID- 9856260 TI - [Physiopathology of atopic dermatitis]. PMID- 9856261 TI - [Physiopathology of leukocytoclastic vasculitis]. PMID- 9856262 TI - [Physiology of erythema multiforme]. PMID- 9856263 TI - [Physiopathology of drug dermatitis]. PMID- 9856264 TI - [Physiopathology of autoimmune bullous dermatoses of the basement membrane]. PMID- 9856265 TI - [Physiopathology of venous leg ulcers]. PMID- 9856266 TI - [Physiopathology of adrogenetic alopecia]. PMID- 9856267 TI - [Physiopathology of infectious dermatitis]. PMID- 9856268 TI - [Physiopathology of sarcoidosis]. PMID- 9856270 TI - [Physiopathology of langerhans-cell histiocytosis]. PMID- 9856269 TI - [Physiopathology of acanthosis nigricans]. PMID- 9856271 TI - [Physiopathology of nevi and nevocytes]. PMID- 9856272 TI - [Habitual shoplifting in multi-infarct syndrome]. AB - The authors report on a 59-year-old male who repeatedly had been apprehended for shoplifting. The behavior only started after he had suffered multiple ischemic strokes. The deviant behavior is explained as a consequence of frontal lobe lesions leading to behavioral disinhibition. Criminal responsibility had to be denied. PMID- 9856273 TI - [Economic crisis and suicide in Geneva: 1991-1995]. AB - The study presented includes all cases of suicide in Geneva from 1991 to 1995. Data reviewed in this article show a decrease of the suicide rate despite of severe unemployment as a concomitant of economic stagnation. This fact may be attributable to the efficiency of local preventive measures in the field of medical care and social welfare. PMID- 9856274 TI - [Routine case of "auto-erotic accident"--surprising turn in evaluation of cause of death after chemical-toxicologic analysis]. AB - A man with indications of autoerotic manipulations was found dead in his flat and a homicide had to be excluded. Diverse material found in the flat suggested that the man was a consumer of cocaine. But the analyses of blood, urine and organs showed high concentrations of amphetamine and it was thought about an unintentionally mix-up with cocaine. A hair analysis was made and only amphetamine was detected. Therefore the utensils (mirror, razor-blade and sniff pipe) belong to a rare nasal consumption of amphetamine. PMID- 9856275 TI - [Possibilities for toxicologic evidence in adipocere formation and postmortem interval of several years]. AB - It is reported on a 31-year-old man, whose dead body was found together with numerous packets of poisons and medicaments in a forest after a post-mortem period of nearly 3 years. Despite advanced skeletization and complete transformation of the still existing residual soft tissues to adipocere, highly toxic concentrations of two heavy metals (cadmium: 0.30 mg/kg; thallium: 0.91 mg/kg) and minor levels of three organic substances (phenobarbitone: 0.32 mg/kg; nordazepam: 0.14 mg/kg; salicylic acid: 0.04 mg/kg) were detected in adipoceratous samples. Even if it is not possible to derive similar blood levels from post-mortem values, the cause and manner of death (suicidal intoxication) could be clarified with the necessary degree of certainty. Few comparable literature reports either deal with exhumations or non-toxic concentrations of substances in morphologically better preserved adipoceratous bodies. Our case demonstrates that toxicological analyses may contribute to the clarification of the cause of death even if advanced adipocere formation with a longer post-mortem interval is present. PMID- 9856276 TI - [Phenomenology of ether administration. With a review of the literature]. AB - A 45-year-old man was found dead lying on a sofa in his house. He had pulled a plastic bag over his head, the bag containing a pad of cotton-wool soaked with diethylether. Different aspects are discussed in this paper including a review of the literature: anamnesis, scene of death, utensils used for inhalation and autopsy findings. There are especially pointed out the results of chemical toxicological analyses in many body compartments. Furthermore are interpreted the manners and the relations of the distribution of ether as well as possible effects on the consciousness. In consideration of other published cases, the analytical results concerning ether are discussed with regard to the presumed cause of death (e.g. asphyxia) and other potential competitive mechanisms. PMID- 9856277 TI - [Death in the milieu of homeless persons. A comparative retrospective analysis of an autopsy sample of different regional population structures (Middle Hessia/Hamburg)]. AB - For an evaluation concerning different regional population and community characteristics, 23 fatalities of homeless individuals were selected from the current autopsy series of the Institutes of Legal Medicine of Giessen/Germany and Hamburg/Germany from 1990 to 1996. The outcome of this retrospective analysis enables to overcome the deficiency of knowledge of causes, manners and circumstances of death among homeless individuals. PMID- 9856278 TI - [Suicidal excess--presentation of an unusual case]. AB - The authors report the case of a suicide pact of two men (22 and 24 years old), which exhibits extraordinary circumstances concerning the personal profile and the suicide procedures. Both had a psychiatric history and were found dead near their severely damaged car, with gaggs in their mouth and a rope tightened around the neck and fixed to the car. Extra gas cans had also been placed in the car. Autopsy confirmed death due to severe trauma and rope strangulation. Toxicological analysis of the blood revealed codeine, dihydrocodeine, paracetamol in high, doxepine in lethal concentrations. According to police investigations, which has been confirmed by medical examination, both men had made arrangements for their suicide selecting a range of methods to kill themselves. PMID- 9856279 TI - [Individual identification of cadaver parts after a bomb explosion using oligonucleotide fingerprinting by (GTG)5]. AB - A blind shell suddenly and unexpectedly exploded, and 20 dismembered human remains were discovered. DNA fingerprint was performed to determine whether the 20 human remains were derived from one person or not. DNA was isolated from each of the remains and digested by the restriction enzyme Hinf I and Hae III and hybridized with the oligonucleotide probe (GTG)5. DNA fingerprint using Hinf I demonstrated the same band pattern in 17 out of the 20 remains. However, in the remaining 3 samples, two novel strange bands were observed. DNA fingerprint using Hae III showed completely identical pattern in all of the remains. PMID- 9856281 TI - Neuronal death in the central nervous system during development. AB - About half the neurons in the brain die at the time when their connections are being formed. This neuronal death is regulated by anterograde and retrograde signals that reflect both electrical activity and the uptake of trophic factors. Our recent data on the isthmo-optic projection indicate that there are in fact two different retrograde signals: a slow-acting survival signal mediated by a neurotrophin, and a fast-acting death signal mediated by calcium entry due to electrical activity in the presynaptic terminals. The developmental roles of the cell death are not well understood, but they appear to include the elimination of aberrant connections. The intracellular mechanisms of the cell death may not always correspond to the apoptotic ones so thoroughly investigated in vitro, because only one of the three morphological types occurring regularly in vivo resembles apoptosis. However, our experiments on retinal ganglion cells indicate that several apoptotic mechanisms apply in this particular in vivo situation: these include an involvement of oxygenated free radicals and glutathione, cell cycle-related events, and probably the synthesis of proteins promoting neuroprotection or cell death. PMID- 9856280 TI - Direct evidence for glutathione as mediator of apoptosis in neuronal cells. AB - Recent evidence has focused attention on the role of oxidative stress in various acute and chronic neurodegenerative diseases. Particularly, a decrease in the level of the powerful antioxidant glutathione (GSH) and death of dopaminergic neurons in substantia nigra are prominent features in Parkinson's disease. The mode of neuronal death is uncertain; however, apoptosis has been hypothesized to be mediated through the induction of free radicals via oxidative pathways. An approach to determine the role of GSH depletion in neurodegeneration and apoptosis was to create a selective modulation of this antioxidant by metabolic manipulations in a clonal cell line of neuronal origin (mouse neuroblastoma NS20Y). Intracellular GSH levels was lowered by inhibiting its biosynthesis with L-buthionine-(S,R)-sulfoximine (BSO), a specific inhibitor of gamma glutamylcysteine synthetase. This treatment led to a GSH depletion of 50% after 1 h and 98% after 24 h. A direct cause/effect relationship between GSH depletion and apoptosis was evidenced in this neuronal cell type. GSH depletion induced the death of NS20Y and promoted nuclear alterations of apoptosis as demonstrated by the in situ staining of DNA fragmentation after 5 days of BSO treatment (by terminal-deoxynucleotide transferase-mediated dUTP-nick end labeling), and the appearance of DNA laddering on agarose gel. These results suggested that redox desequilibrium induced by GSH depletion may serve as a general trigger for apoptosis in neuronal cells, and are consistent with the hypothesis that GSH depletion contribute to neuronal death in Parkinson's disease. PMID- 9856282 TI - Pro-apoptotic effect of the hepatitis B virus X gene. AB - The hepatitis B virus (HBV) is a common human pathogen that causes acute and chronic liver disease. Persistent HBV infection is strongly associated with the development of hepatocellular carcinoma. The contribution of the viral regulatory protein HBx in liver oncogenesis has been supported by our recent studies in a transgenic mouse model, showing that HBx cooperates with c-myc by accelerating the onset of primary liver tumors. Here we show that liver expression of HBx is associated with increased rates of spontaneous apoptosis in liver cells from two different transgenic lines. In transient transfection assays, overexpression of HBx in the established hepatocyte cell line MMHD3 and in human hepatoma cells HepG2 was found to induce apoptosis in a dose-dependent manner. These data suggest that HBx might trigger an apoptotic process in HBV-infected hepatocytes, in turn possibly favoring liver regeneration and accumulation of genetic alterations, ultimately leading to liver cell transformation in chronically infected patients. PMID- 9856283 TI - Cell death induced in lymphocytes expressing the elastin-laminin receptor by excess agonists: necrosis and apoptosis. AB - This manuscript summarizes our experiments carried out during the last years on the expression of the elastin-laminin receptor on human activated lymphocytes and cell death triggered by the activation of this receptor by its agonists, elastin peptides. We could distinguish two types of cell reactions, depending on the elastin peptide concentration added to the cell culture media of lymphocytes. At low concentrations (1-10 micrograms/mL, 1.3-13 x 10(-8) M) of kappa-elastin, there was a stimulation of cell proliferation, elastase biosynthesis and release. As the concentration of kappa-elastin was increased in the culture medium up to 100 micrograms/mL, lymphocyte proliferation and elastase production decreased and the proportion of dead cells increased. Cell death was shown to be due to both apoptotic and non-apoptotic mechanisms. Apoptotic cell death increased with agonist concentration and reached approximately 60% of the lymphocyte population at mg/mL elastin peptide concentrations. This observation was confirmed by the concomitant use of several different methodologies, such as flow cytometry and electron microscopy. The precise nature of the non-apoptotic cell death remains to be established. PMID- 9856284 TI - Fas-mediated apoptosis of hepatic cells. AB - While the fas/fas ligand system has been extensively investigated in immuno competent cells, the place of this system in the physiology and pathophysiology of liver cells remains to be clarified. Although we know that fas is present at the surface of hepatocytes--the main hepatic cells--the role of this membranous protein in physiological conditions is not yet elucidated. However it is the localization of fas on the plasma membrane of hepatocytes which explains why these cells are mainly destroyed by apoptosis--in a picture resembling human fulminant hepatitis--when mice are administered with anti-fas antibodies or fas ligand. It is also established that fas is surexpressed in some human chronic liver diseases, such as those induced by hepatitis B or C virus, a situation which could explain the pathogenesis of some liver lesions occurring during these diseases, such as the apoptosis of hepatocytes in piecemeal necrosis. Finally the fact that caspases, a group of cysteine proteases activated in fas-induced apoptosis, opens the way to inhibition of these enzymes by synthetic peptides and to prevent and treat hepatocyte apoptosis. Demonstration of this possibility has been recently reported in animals presenting fulminant hepatitis induced by anti fas antibodies. PMID- 9856285 TI - The 2-5A system in viral infection and apoptosis. AB - The 2-5A system is an established endogenous antiviral pathway. Interferon treatment of cells leads to an increase in basal, but latent, levels of 2-5A dependent RNase (RNase L) and the family of 2'-5' oligoadenylate synthetases (OAS). Double-stranded RNA, thought to be derived from viral replication intermediates, activates OAS. Activated OAS converts ATP into unusual short 2'-5' linked oligoadenylates called 2-5A [ppp5'(A2'p5')2A]. The 2-5A binds to and activates RNase L which cleaves single stranded RNA with moderate specificity for sites 3' of UpUp and UpAp sequences, and thus leads to degradation of cellular rRNA. During apoptosis, generalized cellular RNA degradation, distinct from the differential expression of mRNA species that may regulate specific gene expression during apoptosis, has been observed. The mechanism of RNA breakdown during apoptosis has been commonly considered a non-specific event that reflects the generalized shut down of translation and homeostatic regulation during cell death. Due to the similar RNA degradation that occurs during both apoptosis and viral infection we investigated the potential role of RNase L in apoptosis. To investigate whether RNase L activity could lead to apoptosis, NIH3T3 cells were transfected with a lac-inducible vector containing the human RNase L gene. Treatment of these cells with isopropylthiogalactoside (IPTG) caused loss of cell viability that was confirmed as an apoptotic cell death by morphological and biochemical criteria. Similarly, specific allosteric activation of endogenous RNase L by introduction of 2-5A directly into L929 cells also induced apoptosis. In L929 cells poly(I).poly(C) treatment in combination with interferon caused an increase in apoptosis whereas neither interferon or double stranded RNA alone altered cell viability. Therefore, increased expression or activation of RNase L causes apoptosis. Inhibition of RNase L, specifically with a dominant negative mutant, suppressed poly(I)Ypoly(C)-induced apoptosis in interferon-primed fibroblasts. Poliovirus, a picornovirus with a single-stranded RNA genome, causes apoptosis of HeLa cells. Expression of the dominant negative inhibitor of RNase L in HeLa prevented virus-induced apoptosis and maintained cell viability. Thus, reduction or inhibition of RNase L activity prevents apoptosis. Both apoptosis and the 2-5A system can provide defense against viral infection in multicellular organisms by preventing production and therefore spread of progeny virus. RNase L appears to function in both mechanisms, therefore, initiation of apoptosis may be one mechanism for the antiviral activity of the 2-5A system. PMID- 9856286 TI - Data of pre-clinical and early clinical trials of acriflavine and hydroxy-methyl ellipticine reviewed, enriched by the experience of their use for 18 months to 6 years in combinations with other HIV1 virostatics. AB - Two virostatics which we discovered in 1990, acriflavine (ACF) and hydroxy-methyl ellipticine (HEL) and shown active on HIV1 resistant to AZT have been introduced into combinations of four virostatics selected among ten available: themselves, plus zidovudine, zalcitabine, didanosine, lamivudine, stavudine, saquinavir, ritonavir, indinavir, the combinations were applied in 3-week sequences, differing from each other by drug rotation. Those which contained ACF may have more often a CD34 decrease than those containing neither ACF nor HEL, and they present more often a CD4 increase. No significant difference as far as side effects or beneficial effects could be detected after 18 months to 6 years, between sequences containing ACF or HEL or both, and sequences not containing any one of them. PMID- 9856287 TI - Thiorphan stimulates clonal growth of GM-CFU in short-term cultures of bone marrow from a healthy donor and from patients with non-Hodgkin lymphoma. AB - Thiorphan, a specific inhibitor of membrane neutral endopeptidase (NEP, EC 3.4.24.11) also known as the common acute lymphoblastic leukemia antigen (CALLA, CD10) was added into short-term clonal cultures of the buffy coat concentrates of human bone marrow obtained from a healthy donor (six experiments) and from ten patients with non-Hodgkin lymphoma (NHL) (eight in complete remission, one in partial remission and one in relapse). Thiorphan concentrations ranged from 10( 5) to 10(-13) M. Nanomolar and higher concentrations of the drug mildly stimulated the granulocyte-macrophage colony-forming unit (GM-CFU) counts in the cultures of normal bone marrow, reaching the significance at 10(-7) M. Meaningful alterations of the GM-CFU counts were noted in 31 of 79 thiorphan-treated cultures of NHL bone marrow (39%). In those cultures the stimulatory effects (33%) outnumbered the inhibitory ones (6%). The stimulatory effects occurred mainly in the bone marrow samples of the patients with highly malignant NHL. The observations are compatible with the idea that the membrane endopeptidase (CALLA, CD10) participates in processes controlling the proliferation and differentiation of hematopoetic cells by cleaving the neuropeptides and related hemoregulatory peptides. PMID- 9856288 TI - Bile immunoglobulins and blockage of biliary endoprosthesis: an immunohistochemical study. AB - BACKGROUND: Endoscopic biliary stenting for pancreaticobiliary malignancy is often limited by recurrent stent occlusion as a result of bacterial biofilm formation and sludge deposition. Bile immunoglobulins are thought to be important in combating biliary sepsis. OBJECTIVES: To investigate whether bile immunoglobulins are involved in the pathogenesis of stent blockage. DESIGN: Immunohistochemical technique was used to study the distribution of bile immunoglobulins, bacteria and sludge in blocked biliary stents. METHODS: Patients with malignant obstructive jaundice were palliated by endoscopic insertion of a 10-FG polyethylene stent into the biliary tract. Blocked stents were retrieved from those who presented with recurrent jaundice and fever. The stents were cross sectionally cut into slices and fixed in formalin. Immunoglobulins were demonstrated by the peroxidase-anti-peroxidase staining procedure using rabbit anti-serum. RESULTS: The central bulk of the stent deposits appeared as an amorphous, structureless material. IgA was found as a rim of dark brown discoloration at the periphery. IgG showed similar distribution and intensity to that of IgA whereas little IgM was detected. CONCLUSIONS: Bile immunoglobulins may facilitate bacterial adhesion, clumping, and hence biofilm formation on the stent surface. PMID- 9856289 TI - Thermal mud-pack as an anti-inflammatory treatment. PMID- 9856290 TI - Recovery of positive future thinking within a high-risk parasuicide group: results from a pilot randomized controlled trial. AB - OBJECTIVES: The research examined: (i) whether high risk parasuicide patients showed a deficit in positive future thinking but no increase in negative future thinking; and (ii) whether such a deficit could be remedied by a brief, manual assisted psychological intervention (manual assisted cognitive-behaviour therapy; MACT). DESIGN: A cross-sectional, mixed model design was used to assess differences between a sample of high risk parasuicide patients and matched controls on future thinking. A longitudinal mixed model design was used to assess changes in future thinking in the different groups over time. METHODS: Parasuicide patients with a history of previous suicidal behaviour and personality disturbance were compared with a matched group of community controls on an adapted fluency measure of future thinking, which measured both quantitative and qualitative aspects of anticipated experiences. Patients were then randomly allocated to either the specific intervention (MACT) or treatment as usual (TAU) and assessed again at 6 month follow-up. RESULTS: Parasuicide patients showed reduced positive future thinking but no increased negative future thinking. Patients who received MACT showed a significant improvement in positive future thinking over the follow-up period whereas the TAU group showed no such improvement. However, interpretation of this finding was made more difficult by the control group also showing a significant improvement in positive future thinking. CONCLUSION: The results confirm that parasuicide patients exhibit a relative deficit in positive future thinking and suggest that this lack of positive future thinking may be remedied, at least partly, by a brief intervention. PMID- 9856291 TI - Reality negotiation in non-depressed and depressed persons. AB - OBJECTIVES: Non-depressed and depressed persons were hypothesized to input and recall self-referential information that was consistent with their self theories. DESIGN: Non-depressed and depressed persons were compared. METHODS: Non-depressed (14) and depressed (14) students chose between positive or negative audio statements, and then recalled the statements. RESULTS: Non-depressed relative to depressed participants: (i) listened more to the positive than the negative tapes; and (ii) fabricated (i.e. incorrectly recalled) less negative messages. CONCLUSION: Partial support is given to reality negotiation processes. PMID- 9856292 TI - Depressed and non-depressed mothers with problematic preschoolers: attributions for child behaviours. AB - OBJECTIVES: This study examines the spontaneous causal attributions made by mothers about their preschool children's problem behaviour and investigates the relationship between causal attributions and maternal depression. DESIGN: Two groups of mothers were compared, a depressed and a non-depressed group, while all women included identified their preschool child as having some problem behaviours. There were 25 women in each group. METHODS: The spontaneous attributions of mothers were assessed from audiotaped interviews using an adaptation of the Leeds Attributional Coding System for the extraction and analysis of attributional statements. RESULTS: Depressed mothers made more spontaneous causal attributions about their children's problem behaviour than did the non-depressed group; and the depressed group perceived these causes as being more stable, more controllable and more personal to the child than their non depressed counterparts. There was also evidence that depressed mothers made more internal attributions about themselves as the cause of their children's problem behaviour than non-depressed mothers. The study indicates that the personal-to child dimension is the attributional variable most strongly associated with depression. CONCLUSIONS: This study provides evidence that depressed mothers have attributional biases when compared to non-depressed mothers. The authors suggest that their attributions may mediate coping responses and hence may influence parenting behaviour. The clinical and research implications of the findings are discussed. PMID- 9856293 TI - The mental health of Muslim mothers in extended families living in Britain: the impact of intergenerational disagreement on anxiety and depression. AB - OBJECTIVES: The study assessed the impact of intergenerational differences of opinion over child rearing on the mental health of Muslim mothers living in extended families. DESIGN: The study adopted a correlational design in an attempt to identify factors that accounted for mental health problems. METHODS: The child rearing attitudes of mothers and grandmothers, mothers' mental health, levels of family acculturation and a range of other background and demographic information was collected from 54 extended families living in two Muslim communities in London using Urdu versions of standard questionnaires. RESULTS: Rates of depression and anxiety among the mothers in the study were high. Grandmothers had more traditional attitudes to child rearing than did mothers. Intergenerational discrepancy over child rearing was more marked in more acculturated families. Discrepancy was associated with higher levels of mothers' anxiety and depression. CONCLUSIONS: The unusually high levels of depression and anxiety displayed by Muslim mothers living in extended families can in part be accounted for by patterns of intergenerational discrepancy. These possibly reflect discordant world views within those families that have been assimilated into the dominant British culture. PMID- 9856294 TI - Dependency and self-criticism in bipolar and unipolar depressed women. AB - OBJECTIVES: The dependency and self-criticism scores of women with histories of either unipolar disorder (N = 74) or bipolar disorder (N = 20) or no psychiatric history (N = 24) were compared to determine the influence of state depression on these personality dimensions. DESIGN: All women completed the Depressive Experiences Questionnaire and the Beck Depression Inventory. Patients were divided into currently depressed and remitted groups. RESULTS: Currently depressed women with unipolar disorder were more self-critical and dependent than non-psychiatric controls; women with unipolar disorder whose depression remitted were more self-critical only. Women in both the depressed and remitted bipolar groups were more self-critical than controls. Depressed bipolar patients did not differ from the controls in their level of dependency; remitted bipolar patients were significantly less dependent than controls. CONCLUSIONS: Dependency needs in depressed women are heavily influenced by mood state. Self-criticism appears to be a-characterological trait in both major depression and bipolar disorder. Women with bipolar disorder in remission report fewer dependency needs than women with no history of psychiatric disorder. The data provide partial support for Blatt's (1974) hypothesis that dependency and self-criticism reflect relatively stable personality dimensions in patients with a mood disorder. PMID- 9856296 TI - A retrospective analysis of expressed emotion (EE) and affective distress in a sample of relatives caring for traumatically brain-injured (TBI) family members. AB - OBJECTIVES: To examine levels of expressed emotion (EE) and affective distress in relatives caring for severely brain-injured family members. DESIGN: The study took the form of a retrospective, observational cross-sectional analysis. METHOD: Data on 28 relatives were reviewed. Each participant completed one affective health-related questionnaire (Leeds Scales for the Self-Assessment of Anxiety and Depression; Snaith, Bridge & Hamilton, 1976) and their EE was rated using the Camberwell Family Interview (CFI; Brown, Birley & Wing, 1972; Vaughn & Leff, 1976, modified for head-injury relatives. Participants were rated as either high or low EE. Comparative statistical analysis examined differences in affective distress between the two groups. RESULTS: Of the 28 participants, 12 were rated high EE and 16 were rated low EE. Levels of anxiety were significantly higher among the high EE group, but levels of depression were not significantly different between the high and low EE groups. A significant correlation was found between EE and carer status, i.e. sole carers or family carers, with sole carers displaying significantly higher levels of EE than family carers. Multiple regression also revealed that carer status was most predictive of EE. CONCLUSIONS: Discussion considers EE as a coping strategy. Further research into the predictive capability of EE as an indicator of affective distress in head injury families is suggested. PMID- 9856295 TI - The London-East Anglia randomized controlled trial of cognitive-behaviour therapy for psychosis. IV: Self-esteem and persecutory delusions. AB - OBJECTIVES: There has been a resurgence of interest in the view that persecutory delusions serve a function of defending self-esteem. An alternative account of levels of self-esteem in individuals with persecutory delusions is that they result from processes similar to those studied in people with depression (i.e. from the occurrence of a range of life experiences and how the individual interprets and copes with them). This study aimed to examine both hypotheses together for the first time, and, as the literature indicates that delusions may not share a common cause, attention was given to the possibility of the presence of subgroups. DESIGN: Data were examined cross-sectionally and longitudinally from a randomized controlled trial of cognitive behaviour therapy for 60 people with drug-resistant psychosis. METHOD: The study is based on the initial assessment of all participants on self-esteem, delusional conviction and a large number of demographic, clinical and cognitive measures. Longitudinal analyses were also carried out, and are reported separately for those who received the therapy intervention and those in the control group. RESULTS: Almost three quarters of participants with persecutory delusions reported low self-esteem. Changes over time in total self-esteem correlated with changes in measures of mood and social functioning, but not conviction in persecutory delusions. The individuals who initially had normal levels of self-esteem displayed a different pattern of results from the majority of participants. CONCLUSION: Low self-esteem in people with drug-resistant persecutory delusions is common and, in most cases, can best be understood in terms of normal emotional processes. There was evidence that the majority of persecutory delusions do not fit either strong or weak formulations of the delusion-as-defence explanation and that there may be subgroups with differing aetiologies. These results need to be replicated, and extended to groups in which symptoms are not resistant to medication. PMID- 9856297 TI - Learning disability care staff's emotional reactions to aggressive challenging behaviours: development of a measurement tool. AB - OBJECTIVES: To describe the development of a rating scale measure of caregivers' emotional reactions to aggressive challenging behaviour, and to report preliminary psychometric data for the scale. DESIGN: Using previous research on staff working with people with learning disabilities, a range of likely emotional reactions to challenging behaviours were selected for possible inclusion in a rating scale measure. METHODS: A total of 83 care staff from 23 community residences for people with learning disabilities rated their recent emotional reactions to aggressive challenging behaviour using 18 emotion items. A further sample of 18 care staff participated in a test-retest study after the initial stages of scale development. RESULTS: A rating scale was developed on the basis of a factor analysis and further item analysis. The measure has two subscales: feelings of depression/anger (10 items), and feelings of fear/anxiety (5 items). The subscales have a high internal consistency, good test-retest reliability and are relatively unaffected by social desirability biases. CONCLUSIONS: The emotional reactions to aggressive challenging behaviour scale has excellent face and construct validity, and other preliminary psychometric data are promising. The scale has a number of potential research and clinical applications. PMID- 9856298 TI - Continuous norming: improved equations for the WAIS-R. AB - OBJECTIVES: To provide more satisfactory continuous norming equations for calculation of WAIS-R IQ figures than are given in Zachary & Gorsuch (1985), and to provide additional equations applicable to subtest scores. DESIGN: Psychometric analysis of published normative data. METHOD: Multiple regression analysis of relevant data concerning IQ scales and subtest scores as tabulated in Wechsler (1981), involving appropriate transformations of scores and age, followed by relevant analyses of differences between calculated and tabulated values. RESULTS: The derived equations perform relatively well. Computational details are provided. CONCLUSIONS: Continuous norming procedures have recognised practical and theoretical advantages. Equations presented here fit tabulated WAIS R data more closely than do the previous equations; the general method and transformations used may be applicable to other comparable normative data sets. PMID- 9856299 TI - The Modified Card Sorting Test: test-retest stability and relationships with demographic variables in a healthy older adult sample. AB - OBJECTIVES: To investigate the test-retest stability of a standardized version of Nelson's (1976) Modified Card Sorting Test (MCST) and its relationships with demographic variables in a sample of healthy older adults. DESIGN: A standard card order and administration were devised for the MCST and administered to participants at an initial assessment, and again at a second session conducted a minimum of six months later in order to examine its test-retest stability. Participants were also administered the WAIS-R at initial assessment in order to provide a measure of psychometric intelligence. METHODS: Thirty-six (24 female, 12 male) healthy older adults aged 52 to 77 years with mean education 12.42 years (SD = 3.53) completed the MCST on two occasions approximately 7.5 months (SD = 1.61) apart. Stability coefficients and test-retest differences were calculated for the range of scores. The effect of gender on MCST performance was examined. Correlations between MCST scores and age, education and WAIS-R IQs were also determined. RESULTS: Stability coefficients ranged from .26 for the percent perseverative errors measures to .49 for the failure to maintain set measure. Several measures were significantly correlated with age, education and WAIS-R IQs, although no effect of gender on MCST performance was found. CONCLUSIONS: None of the stability coefficients reached the level required for clinical decision making. The results indicate that participants' age, education, and intelligence need to be considered when interpreting MCST performance. Normative studies of MCST performance as well as further studies with patients with executive dysfunction are needed. PMID- 9856300 TI - Anti Rh 17 in a multigravida with Rh-D-/[symbol: see text] genotype resulting in haemolytic disease of the newborn. PMID- 9856301 TI - The influence of norethisterone acetate on urinary urodilatin excretion in postmenopausal women. AB - We have previously found that transdermal estradiol application significantly stimulated the urinary excretion of urodilatin, a newly discovered renal peptide with diuretic properties. It is well established that the addition of progestogen is necessary in hormone replacement therapy in women with an intact uterus. This study was designed specifically to examine the effect of progestogen norethisterone acetate (NETA) in postmenopausal women. NETA given alone orally in dosages of 1 mg/d and 2 mg/d for 10 days, as used for the progestogen-challenge test, did not increase urodilatin excretion. NETA in combination with estradiol, administered orally and transdermally in the second half of a 4-week estradiol treatment cycle, did not significantly change urodilatin excretion. The results of the present study indicate that NETA addition to estrogen replacement therapy may antagonize the stimulating estradiol effect on urodilatin production as registered in our earlier study. PMID- 9856302 TI - Intrauterine ectopic pregnancy. A case report. AB - A case of vaginal bleeding occurring in a patient with a cervico-isthmic pregnancy located in a previous caesarean section scar was treated with curettage and systemic methotrexate. Metrorrhagia and uterine rupture are severe complications of this kind of ectopic pregnancy and sometimes require demolitive surgery, still today. Early ultrasonographic diagnosis is necessary to allow conservative treatment. PMID- 9856303 TI - Ruptured corpus luteum cyst of the ovary following non-perforated acute appendicitis. Three case reports. PMID- 9856304 TI - Artificial endometrial preparation for oocyte donation using synthetic estrogen and progestogen. AB - OBJECTIVE: To evaluate the outcome of an oocyte donation program using synthetic estrogen and progestational agents for uterine preparation. METHODS: Conjugated estrogen, 1.25 mg per day at increasing doses and dydrogesterone, a synthetic progestogen, 30 mg per day were used for uterine priming. All embryo transfers were done on day 2 of progestogen supplementation. RESULTS: The pregnancy rates were 38% per embryo transfer. The ongoing pregnancy rate was 31% with an abortion rate of 20%. CONCLUSION: Endometrial preparation in an oocyte donation program using orally administered synthetic estrogen and progestogen gives pregnancy rates comparable to those reported with natural products. PMID- 9856305 TI - A new approach to laparoscopic treatment for interstitial pregnancy. PMID- 9856306 TI - Fetal ovarian cysts: prenatal diagnosis and management. Report of two cases and review of literature. AB - The Authors report two cases of antenatally diagnosed fetal ovarian cysts. In the first case the cysts underwent spontaneous resolution. In the second case the newborn was submitted to adnexectomy for cyst torsion. A review of the literature is reported. PMID- 9856307 TI - Congenital infection by human parvovirus B19 ascites-anaemia. AB - A case of intrauterine infection by human parvovirus B19 (HPV B19) manifested as ascites during pregnancy is presented. Ascites was diagnosed by ultrasound at 27 weeks' gestation. A caesarean section was performed at 37 weeks'. owing to affected mobility of the fetus. A pale, female infant with low haemoglobin and bradycardia was delivered. Polymerace Chain Reaction (PCR) lab tests revealed that the mother and the fetus were infected by HPV B19. The neonate was born with low haemoglobin (Hb = 10 g/dl) and with ascites; it was discharged in good general condition 50 days after delivery. PMID- 9856308 TI - Rectosigmoid endometriosis: diagnosis and surgical management. AB - The recurrence of endometriosis varies from 6% to 10% and, among the non gynaecological sites, the bowel is involved in 12%-37%. Various symptoms, such as dysmenorrhea, dyspareunia, chronic pelvic pain, diarrhoea, constipation, cyclic rectal bleeding, colic-abdominal pain up to intestinal occlusion characterize this pathology. Surgery seems to be the best treatment especially for gastrointestinal symptoms; conservative surgery should be performed, particularly in young patients. Four cases of intestinal endometriosis were reevaluated. PMID- 9856309 TI - Spontaneous pregnancy occurring in a donor oocyte cycle: a case report. PMID- 9856310 TI - Pulmonary edema as an acute complication of ritodrine therapy in the presence of maternal intrauterine infection. PMID- 9856311 TI - Is there a safe anticoagulation protocol for pregnant women with prosthetic valves? AB - We studied the outcome of 41 pregnancies in an attempt to identify an appropriate and safe anticoagulant regimen for pregnant women with cardiac valve prosthesis. The cumulative number of events: (neonatal death, premature deliveries, intrauterine fetal death and spontaneous abortion) was 66.6%, 20%, 12.5% and 20% in the Coumadin, Fraxiparine, Heparin and porcine valve groups, respectively. We conclude that in pregnant women with mechanical heart valves, low molecular weight heparin therapy is a superior strategy compared to warfarin. PMID- 9856312 TI - Advantages of spinal anesthesia in abdominal gynecologic surgery. AB - The advantages obtained in vaginal surgery and caesarean section using spinal anesthesia led us to test this anesthesia to verify feasibility, problems and advantages in abdominal surgery. Spinal anesthesia was performed in 60 patients between 21 and 87 years of age. Thirty-seven total abdominal hysterectomies with or without adnexectomy, 5 laparotomic miomectomies, 3 adnexectomies, 5 colposacropexies, 2 hysterectomies with lymphadenectomy, 7 Burch colposuspension with or without hysterectomy and 1 laparoscopy for sterilization were performed. No significant problems during surgery and the postoperative period were observed. Resumption of the different physiologic functions were more rapid, hospital stay shorter and compliance greater than with general anesthesia. PMID- 9856313 TI - The effect of 61 days of combined iron (Chemiron) and single iron therapy on haemoglobin, packed cell volume, platelets and reticulocytes during pregnancy. Preliminary report. AB - A 61-day short-term prospective study comparing the efficacy of Chemiron capsules (Ferrous Fumarate 300 mg, Folic Acid 5 mg, Vitamin B12 10 ug, Vitamin C 25 mg, Magnesium sulphate, 0.3 mg and Zinc Sulphate 0.3 mg) with conventional ferrous gluconate and folic acid administration has shown that both haematinic regimens improve haematological indices in normal pregnant women. A significant rise in haemoglobin was seen in the Chemiron group on days 5 and 19 whereas a significant increase in packed cell volume was observed on days 5, 19 and 61. There were no significant differences between the mean value of platelets during the therapy period in either group. A significant rise in the mean reticulocyte values could be demonstrated in the Chemiron group throughout the study period. This study further shows that Chemiron has a better haematological effect than Ferrous Gluconate at the dosage used. PMID- 9856314 TI - Rupture of the pregnant uterus--a 21-year review. AB - OBJECTIVES: To review the incidence of ruptured uteri at the Women's Hospital, Hamad Medical Corporation (HMC) and to analyze the causative factors of uterine rupture with a view to its prevention and to highlight the management approach taken to preserve the patient's reproductive potential. METHODS: Case notes were reviewed for all patients (except for eight which were not available) with a ruptured uterus at the Women's Hospital in Doha for a period of 21 years from 1 July 1977 to 30 June 1997, relevant data relating to the clinical features characteristics of labor, operative procedures, and maternal perinatal outcome were assessed. RESULTS: There were 31 cases of ruptured uteri. The incidence of ruptured uteri was calculated to be 0.017%, 23 cases being available for the study. Ten cases (43.5%) occurred in patients with previous cesarean scars, while 13 cases (56.5%) were grand multiparous (para 5 or more). In 10 cases (43.5%) uterine rupture was associated with oxytocin use, and four cases (18.2%) were associated with PGE2 use. The ruptures occurred in the lower segment of 19 cases (90.9%). Fetal heart abnormalities were observed in all cases except one in which the uterus ruptured during labor. Abdominal hysterectomy was performed in 15 cases (65.2%). The remaining eight patients had suture repair, two of them had suture repair with sterilization and the other six cases (26.1%) had suture repair without sterilization. Five of them became pregnant and were delivered by cesarean section. CONCLUSIONS: Even though rupture of the uterus was rare in our study, its occurrence should be suspected when there are sudden fetal heart abnormalities during labor or unexpected postpartum shock. Suture repair should be considered whenever possible in order to maintain the patient's future fertility. PMID- 9856315 TI - Association of genital mycoplasma colonization with low birth weight. AB - OBJECTIVE: This study was conducted to document the prevalence of maternal genital tract colonization by Ureaplasma urealyticum and Mycoplasma hominis, and to assess its association with low birth weight (LBW) and prematurity. METHODS: The high vaginal swabs of pregnant women in spontaneous labor after 26 weeks of gestation were cultured for U. urealyticum and M. hominis. Clinical details and perinatal outcomes including birth weight and gestation were documented. RESULTS: Of a total of 303 women enrolled, 148 (48.8%) had positive vaginal cultures for U. urealyticum, while only five (1.6%) grew M. hominis. The mean birth weight and the incidences of LBW and preterm neonates among ureaplasma positive and ureaplasma negative mothers were statistically comparable. CONCLUSIONS: U. urealyticum emerged as a common inhabitant of the lower genital tract of women in labor, being present in nearly half of them. Its presence was not a risk factor of LBW or prematurity. Maternal colonization with M. hominis was uncommon. PMID- 9856316 TI - Vascular cell adhesion molecule-1 (VCAM-1) and leukocyte activation in pre eclampsia and eclampsia. AB - OBJECTIVE: To evaluate the levels of VCAM-1 in pre-eclampsia/eclampsia as a possible marker of leukocyte activation and endothelial damage. METHODS: We performed a case-control study on 25 healthy pregnant women and 35 patients diagnosed as pre-eclampsia or eclampsia which were randomly selected. Peripheral venous blood samples were obtained and serum levels of VCAM-1 were measured by enzyme-linked immunoassay (ELISA). RESULTS: In pre-eclampsia/eclampsia, VCAM-1 levels were higher than in normal pregnancy. Serum concentrations of VCAM-1 were significantly higher in severe pre-eclampsia and eclampsia compared to mild pre eclampsia or healthy controls. ROC analysis detected that VCAM-1 > or = 450 ng/ml had a sensitivity of 0.79 and a specificity of 0.90 in detecting severe pre eclampsia and eclampsia. CONCLUSION: This is the first study to correlate VCAM-1 levels with severity of disease in pre-eclampsia. Our findings indicate that increasing levels of soluble VCAM-1 are present in the circulation of patients with severe pre-eclampsia/eclampsia compared to mild pre-eclampsia or healthy pregnant women. Elevated VCAM-1 levels may represent a possible mechanism by which endothelial cells attract leukocytes and cause endothelial cell damage. PMID- 9856317 TI - Persistent subchorionic hematoma with clinical symptoms until delivery. AB - OBJECTIVE: This study was performed retrospectively to clarify the clinical features of persistent subchorionic hematoma with clinical symptoms until delivery. METHODS: We studied all the 22 cases of persistent subchorionic hematoma with symptoms until delivery from 4763 singleton pregnant women delivered in our hospital from 1985 to 1996 inclusive to investigate their clinical course and pregnancy outcome. RESULTS: The initial symptom of subchorionic hematoma was vaginal bleeding and/or uterine contractions. The onset of the symptoms showed two peaks at 9-11 and 30-31 weeks of gestation. The pregnancy outcome was spontaneous abortion in three cases (13.6%), premature labor in 17 cases (77.3%) or full-term delivery in two cases (9.1%). Tocolysis was needed in 17 of the 22 cases (77.3%), of which 16 cases failed to prevent premature delivery. CONCLUSION: Persistent subchorionic hematoma with clinical symptoms until delivery is likely to result in abortion or premature labor, because that may be thought to be a severe type. PMID- 9856318 TI - Factors affecting postoperative pregnancy rate after endoscopic management of large endometriomata. AB - OBJECTIVE: To identify factors influencing postoperative pregnancy rate in women with extensive endometriosis and large endometriomata as the only identified cause of infertility that were treated by laparoscopy. METHOD: Sixty-four infertile patients with endometriomata (> or = 3 cm) and no other apparent cause of infertility. The latter were removed by operative laparoscopy. Life table calculations, the Student's t-test and the chi2 test were used where appropriate. RESULT: Thirty-four patients (53%) became pregnant during the 2-year follow-up period. A significantly increased pregnancy rate was found for the first year compared to the second (76 vs. 24%). The existence of adhesions affected adversely the outcome of the operation only as far as early achievement of pregnancy is considered. The number and size of endometriomata and the existence of peritoneal implants have not been found to affect pregnancy rates. The severity of the disease did not affect pregnancy rate, but in the cases with moderate disease most of the pregnancies were achieved during the first postoperative year. The duration of infertility was significantly associated only at the 10% level with decreased pregnancy rates. CONCLUSION: Extensive endometriosis with large endometriomata can be safely and effectively treated with laparoscopy using the traditional laparoscopic tools providing the infertile patient with a high chance to conceive in a relatively short period of time postoperatively. PMID- 9856319 TI - Uterine balloon therapy to treat menorrhagia. AB - OBJECTIVE: To study the clinical efficacy, safety and acceptability of the thermal balloon endometrial ablation (TBEA) in patients with dysfunctional uterine bleeding. METHODS: Thirteen patients with DUB who did not respond to medical treatment and dilatation and curettage consented to a trial of TBEA with EASY model of balloon catheter by Gynecare. All procedures were done under intravenous (i.v.) sedation and paracervical block. Patients were mostly discharged within 48 h. Follow-up of 2-19 months is reported. Transvaginal ultrasound and hysteroscopy were performed in six patients after 6 months of TBEA. RESULTS: Twelve patients (92.3%) reported a significant reduction in bleeding. Two patients (15.4%) experienced amenorrhea. Only two patients (15.4%) underwent subsequent hysterectomy, one for persistent menorrhagia and the other for severe pelvic pain. In successful patients of TBEA, transvaginal ultrasonography revealed marked reduction in endometrial thickness and no endometrium was visible in one patient who had developed amenorrhea. In successful cases follow-up hysteroscopy revealed scarring in more than two-thirds of the endometrium. CONCLUSIONS: Thermal balloon endometrial ablation is a safe, simple, effective, easy and minimally invasive procedure which can be done under i.v. sedation and paracervical block in an office setting. It has several advantages over hysterectomy, including preservation of the uterus, avoidance of surgical incision and potential to perform on an outpatient basis. It is a good alternative to hysteroscopic endometrial ablation with a comparable success rate with minimal risk and no limiting factors except the cost of the balloons. However, large scale randomized controlled studies are needed with TBEA and other endometrial ablation procedures. PMID- 9856320 TI - Symptoms and signs in single and mixed genital infections. AB - OBJECTIVE: To compare symptoms and signs in women with single and mixed genital infections. METHODS: The study population comprised 996 apparently healthy women. Gynecological symptoms and signs were looked for and diagnostics for the most prevalent gynecological infections were made. RESULTS: When co-infections were excluded, chlamydial infections, bacterial vaginosis and cervical human papillomavirus infections were associated with a fishy malodor; for the two former conditions an easily bleeding ectopy was also found. Vaginal candidosis showed characteristic symptoms and signs. Genital warts were associated with dysuria, general and lower abdominal pain. Out of 494 women with a genital infection, 112 (22.7%) had a mixed infection, which in some cases influenced symptoms and signs. CONCLUSION: Many women who consider themselves gynecologically healthy, may nevertheless harbor one or more infectious agents. The need to exclude multiple infections is obvious. Positive predictive values were for specific symptoms and signs were generally low. PMID- 9856322 TI - Management of septate uterus by flexible hysteroscopy and Nd:YAG laser. AB - OBJECTIVE: To evaluate the efficiency of flexible Nd-YAG laser hysteroscopic metroplasty. METHODS: We present a retrospective study of 17 patients treated for septate uteri between 1990 and 1995. The indication of hysteroscopy was recurrent abortion in nine cases and eight with primary infertility with indication of IVF in another five cases. We did not prescribe any pretreatment. A flexible hysteroscope with a 100-W Nd:YAG laser was used with glycine-controlled flow. The septum was divided by the laser after exploration of the cavity. We proposed a control office hysteroscopy 2 months after surgery to prevent uterine adhesions. RESULTS: There was no complication. Twelve patients conceived with 10 live births at term and two spontaneous abortions. Four are still infertile, two were lost from the study and two no longer desired pregnancy. Resectoscopic studies showed the same results. CONCLUSION: The advantages of this method are the excellent ergonomic properties of the fibroscope and the safety of the laser. We suggest that metroplasty does not improve the pregnancy rate but only the pregnancy outcome of these patients. However, for those with the indication of IVF the procedure seems useful in avoiding abortion. PMID- 9856321 TI - Aspiration and microscopy in the diagnosis of ectopic pregnancy. AB - OBJECTIVE: To compare Karman cannula aspiration followed by dissecting microscopy with suction curettage and permanent histology in obtaining and identifying chorionic villi. METHODS: Karman cannula aspiration was performed before standard curettage for failed intrauterine pregnancies (N=22) or possible ectopic gestation (N=24). Dissection microscopy for chorionic villi was performed on aspirates before submission for permanent histology. Sensitivity, specificity, positive and negative predictive value of each method in obtaining and identifying villi was determined. RESULTS: Overall, all methods were only moderately sensitive in detecting chorionic villi (50-76%). If failed intrauterine pregnancies were excluded, all methods had poor sensitivity (25 64%). However, if villi were detected, the positive predictive value of all methods was high (> 80%). CONCLUSIONS: Karman cannula aspiration followed by dissecting microscope examination or permanent histology may offer an alternative to traditional curettage in the diagnosis of ectopic gestations. A larger trial to validate these findings seems justified. PMID- 9856323 TI - Prognosis-predicting system based on factors related to survival of cervical carcinoma. AB - OBJECTIVE: To establish a different category system based on grouping of the risk factors in patients with cervical smear. METHODS: Univariate and multivariate analyses of factors associated with survival time were performed in 443 patients with stage Ib-IIa squamous cell carcinoma (SCC) from 1985 to 1989. RESULTS: Univariate analysis showed that parametrial extension, bulky tumor size (> or = 4 cm), uterine body involvement, poor differentiation and pelvic lymph node (LN) metastasis were prognostically significant. However, by multiple regression method, only LN metastasis, deep stromal invasion (DSI), and poor differentiation (PD) were significantly related to the patient's survival time with risk ratios of 2.78, 2.61 and 1.52, respectively. A prognosis-predicting system was established on the basis of these simplified factors: groups of high, intermediate, low and very low-risks, obtained survival rates of 93-96%, 77%, 51% and 25-37%, respectively. CONCLUSIONS: Using this simple model, cervical carcinomas can be classified prognostically for predicting 5-year-survival rates, and for risk-guided therapy in the future. PMID- 9856324 TI - Abnormal urodynamic findings after radical hysterectomy or pelvic irradiation for cervical cancer. AB - OBJECTIVE: To assess urodynamic study results in patients with cervical cancer who had received radical hysterectomy or pelvic irradiation or radical hysterectomy with pelvic irradiation. METHODS: Forty-two patients with stage IB cervical cancer after radical hysterectomy (group A), 11 patients at stage IB or IIA after pelvic irradiation (group B), 15 patients at stage IB or IIA after both radical hysterectomy and pelvic irradiation (group C) and 17 patients at stage IB before treatment (group D) as control were recruited for urodynamic examination. The evaluations for each case included a 20-min pad test, uroflowmetry, both filling and voiding cystometry, and stress urethral pressure profile. ANOVA method with Bonferroni test and Pearson chi2-test were utilized for statistical analysis. RESULTS: The mean ages in sequential groups A, B, C and D were 52.9 +/- 10.2, 62.5 +/- 13.5, 49.8 +/- 11.7 and 49.4 +/- 12.5 years (P = 0.02), respectively. The occurring frequency of either detrusor instability or low bladder compliance was 57%, 45%, 80% and 24%, respectively. Each group revealed decreased bladder capacity as 268.4 +/- 102.8, 164.1 +/- 62.9, 233.5 +/- 73.9 and 293.0 +/- 47.2 ml (P < 0.0001). However, the frequency of abdominal strain voiding was 100% in groups A, B and C as compared to 0% in group D (P < 0.01), and the frequency of abnormal residual urine (> 50 ml) was 41%, 27%, 40% and 24%. Although each case showed a poor pressure transmission ratio (< 100%), the frequency of positive pad test in each group was 81%, 46%, 100% and 18% (P < 0.001). The functional urethral length decreased in each group and was 2.6 +/- 0.8, 2.3 +/- 0.8, 2.5 +/- 0.8 and 2.9 +/- 0.6 cm, but there were no significant differences in maximal urethral pressure or urethral closure pressure among the four groups. CONCLUSIONS: Our data show that abnormal urodynamic findings pre exist in patients with cervical cancer before treatment especially in bladder storing function, and that these findings may worsen, or that new abnormal findings may happen after radical hysterectomy or pelvic irradiation, or both. PMID- 9856325 TI - Trends and causes of maternal mortality in Kazakhstan. AB - OBJECTIVE: The aim of this study was to determine trends in the rate and causes of maternal mortality in Kazakhstan. METHODS: Maternal deaths occurring between 1960 and 1996 in the 19 oblast (administrative regions) of Kazakhstan were documented and analyzed. RESULTS: After an initial phase of decrease in the period 1960-1980, the level of maternal mortality in Kazakhstan has been stable at 70-80 maternal deaths per 100,000 live births. Maternal deaths due to hypertension and abortion complications have not substantially decreased. Over the last 10 years, illegal abortions have accounted for most Kazakh maternal deaths resulting from abortion complications. CONCLUSION: To reduce the maternal death rate, the Kazakh health authorities and health workers (obstetricians, midwives, health educators) must develop and rapidly implement maternal health and family planning policies. PMID- 9856326 TI - Trends in maternal mortality in Ilorin, Nigeria 1987-1996. PMID- 9856327 TI - Vacuum extraction for operative vaginal delivery. PMID- 9856328 TI - Labor and delivery of patients with spinal cord injury. PMID- 9856329 TI - Shorter ultrasonic femur lengths in Hmong fetuses. PMID- 9856330 TI - ACOG educational bulletin. Viral hepatitis in pregnancy. Number 248, July 1998 (replaces No. 174, November 1992). American College of Obstetricians and Gynecologists. AB - Hepatitis A is an uncommon complication of pregnancy and is not associated with perinatal transmission. In contrast, hepatitis B virus infection is more common and clearly poses a serious risk to the household contacts and neonates of infected mothers. Accordingly, all pregnant women should be tested for hepatitis B virus. Universal vaccination of all neonates with hepatitis B vaccine is now recommended. Infants delivered to HBsAg seropositive mothers also should receive HBIG and vaccination immediately after birth. Hepatitis E is extremely rare in the United States and is quite similar to hepatitis A, although perinatal transmission does occur with hepatitis E. Hepatitis C and D, which are transmitted parenterally and by sexual contact, have been associated with vertical transmission. No immunoprophylaxis currently is available for neonates of mothers with hepatitis C or E virus. Immunization against hepatitis B is protective against vertical transmission of hepatitis D. PMID- 9856331 TI - ACOG committee opinion. Hepatitis virus infections in obstetrician-gynecologists. Number 203, July 1998 (replaces No. 199, February 1998). Committee on Gynecologic Practice. American College of Obstetricians and Gynecologists. PMID- 9856332 TI - ACOG committee opinion. Institutional responsibility to provide legal representation. Number 204, July 1998. Committee on ethics. American College of Obstetricians and Gynecologists. PMID- 9856333 TI - ACOG criteria set. Cesarean delivery. Number 35, July 1998. Committee on Quality Assessment. American College of Obstetricians and Gynecologists. PMID- 9856334 TI - ACOG criteria set. Quality evaluation and improvement in practice. Cone biopsy of cervix. Number 32, April 1998. Committee on Quality Assessment. American College of Obstetricians and Gynecologists. PMID- 9856335 TI - The ligation systems for ubiquitin and ubiquitin-like proteins. AB - Ubiquitin (Ub) is a highly conserved small protein present universally in eukaryotic cells, which is covalently attached to substrate proteins by a cascade system, consisting of activating (E1), conjugating (E2), and/or ligating (E3) enzymes. The modification of cellular proteins with Ub targets them for degradation by a large multisubunit protease, called the 26S proteasome. The unexpected existence of many genes encoding E2 and E3 reveals that a number of distinct Ub-ligating pathways operate for selective proteolysis in cells, implying its involvement in divergent biologically important processes. Currently, it becomes clear that a set of novel molecules with a structural similarity to Ub, called Ub-like proteins (Ubls), is present in various eukaryotic cells. They are divided into two subclasses: type-1 Ubls with small sizes, such as SUMO1 and NEDD8, that are ligated to target proteins in a fashion similar, but not identical, to the ubiquitination pathway, and another type-2 Ubls that contain Ub-like structure in a variety of different classes of large proteins having apparently distinct functions, such as Rad23, Elongin B, and Parkin. Ub and type-1 Ubls are central players consisting of a new type of post translational protein-modifying system, although the significance of type-2 Ubl remains obscure. PMID- 9856336 TI - Expression of excitation-contraction coupling proteins during muscle differentiation. AB - The goal of this study was to characterize three major excitation-contraction (E C) coupling proteins: ryanodine receptor [RyR, the calcium release channel in the sarcoplasmic reticulum (SR)], dihydropyridine receptor (DHPR, the voltage-gated L type calcium channel in the transverse tubule) and SR Ca2+-ATPase (SERCA, the calcium pump in the SR) in the differentiating primary cultures of rat skeletal and cardiac muscle cells. The expression levels of these proteins were determined by ligand binding assays and/ or immunoblottings along with an examination of the morphological changes in differentiating muscles by phase-contrast microscopy. In the skeletal cells, the expression levels of the E-C coupling proteins generally increased in the course of differentiation with the peak expression at the 9th day of culture. Simultaneous with the increased expression of the proteins, the myoblasts elongated, followed by alignment and fusion of the cells to form multinucleated myotubes. In the cardiac cells, on the contrary, the peak expression levels of DHPR, SERCA and RyR were reached within 2, 4, and 7 d of culture, respectively. Alignment of the cardiac muscle cells and spontaneous contraction occurred as early as several h after plating. These results suggest that expression patterns of E-C coupling proteins are different between skeletal and cardiac muscles, and that DHPR could play an important role in Ca2+ metabolism during the early stages of differentiation. PMID- 9856337 TI - Son of sevenless binds to the SH3 domain of src-type tyrosine kinase. AB - To identify molecules which bind to the SH3 domains of p56lck, we screened a mouse T-cell lymphoma cDNA library using the yeast two-hybrid system. As a result, we obtained several positive clones including the Son of Sevenless gene which encodes a mammalian homolog of Drosophila Ras GDP/GTP exchange factor. In a subsequent analysis with the yeast two-hybrid system, Sos associated only with the constitutively active form of p56lck (F505) but not with wild type p56lck (Y505), indicating the requirement for an active conformation of p56lck for binding to Sos. Subsequently, we have demonstrated in vitro that the SH3 domain of p56lck as well as the proline-rich sequences of Sos are responsible for this association. In addition, the proline-rich domain of Sos also bound to the SH3 domains of other src-type tyrosine kinases, src and fyn, but not to that of PLC gamma. More importantly, the p56lck SH3-Sos interaction was enhanced by serum stimulation, suggesting the possibility that the direct interaction between p56lck SH3 and Sos may contribute to the regulation of the Ras pathway. PMID- 9856338 TI - Cloning and characterization of ribulose bisphosphate carboxylase gene of a carboxydobacterium, hydrogenophagea pseudoflava DSM 1084. AB - The ribulose bisphosphate carboxylase/oxygenase rbcL and rbcS genes of a carbon monoxide-oxidizing bacterium, Hydrogenophaga pseudoflava DSM 1084, were cloned and sequenced. The cloned rbcL and rbcS genes had open reading frames of 1422 and 351 nucleotides encoding RbcL and RbcS with calculated molecular masses of 52,689 and 13,541, respectively. The known active site residues in other RbcL proteins were conserved in the H. pseudoflava proteins. The H. pseudoflava RbcS protein lacked the 12-residue internal sequence found in the plant enzymes. The 2 genes were separated by a 134 bp intergenic region and cotranscribed as a 2.0 kb rbcLS mRNA. Novel two perfect 9 bp direct repeats overlapping with two dyad symmetries were found in the rbcLS promoter region. PMID- 9856339 TI - Matrix attachment region sequences enhanced the expression frequency of a whey acidic protein/human lactoferrin fusion gene in the mammary gland of transgenic mice. AB - To elevate the expression frequency of transgenes in transgenic mice, the chicken lysozyme matrix attachment region (MAR) sequence was used by combining it with a transgene. The whey acidic protein (WAP) promoter/human lactoferrin (hLF) cDNA fusion transgene (pWL) was connected to the chicken lysozyme MAR sequence at its 5'-end (pMWL). While only two of three mice became transgenic from the pWL vector expressed hLF, all seven mice from the pMWL vector expressed the transgene in their lactating mammary glands. To evaluate the effect of lactogenic hormones on transgene expression, experiments with the primary culture of transgenic mammary explants were performed. It was revealed that the expression of transgenes was slightly increased by insulin plus dexamethasone or insulin plus prolactin treatment. However it was not increased by insulin, dexamethasone or prolactin (IDP) treatment alone. In contrast, the endogenous WAP gene was expressed only in the IDP treated group. These results demonstrate that MAR sequences are effective in improving the expression frequency of transgenes in transgenic mice although the developmental and hormonal regulations are not the same as those of the endogenous WAP gene. PMID- 9856340 TI - Molecular cloning and analysis of transcription initiation in the Anagrapha falcifera multiple nucleocapsid polyhedrosis virus polyhedrin gene. AB - Using Autographa californica multiple nucleocapsid polyhedrosis virus (AcMNPV) polyhedrin to probe the Southern blots of Anagrapha falcifera multiple nucleocapsid polyedrosis virus (AfMNPV), we identified the location of the AfMNPV polyhedrin gene within the 7.2 kb EcoRI fragment. The 7.2 kb EcoRI fragment of AfMNPV was cloned and the nucleotide sequences of the polyhedrin coding region and its flanking regions were determined. Nucleotide sequence analysis indicated the presence of an open reading frame (ORF) of 735 nucleotides (nt) which could encode 245 amino acids with a predicted molecular mass of 29 kDa. The nucleotide sequences within the coding region of the AfMNPV polyhedrin shared 80% similarity with the polyhedrin gene from AcMNPV but were most closely related to Bombyx mori NPV with 92% sequence identity. The size of the AfMNPV polyhedrin mRNA, determined by the Northern blot analysis, was estimated to be 1200 nt. The consensus promoter sequence (ATAAG) for the baculovirus very late gene was also observed. Two degenerate poly(A) tailing signals were found immediately downstream of the translational stop codon. The transcription initiation site, mapped by primer extension analysis, was found to be at T located 24 nt upstream from the A of the translation initiation codon. This site is located 26 nt downstream from the second A of the consensus TAAG, the transcription initiation site of most other NPVs. PMID- 9856341 TI - Association between apolipoprotein(a) polymorphism and Lp(a) levels in Koreans. AB - In view of the clinical importance of lipoprotein(a) [(Lp(a)] in coronary artery disease (CAD), we investigated the apo(a) size polymorphism, sequence polymorphisms of five base repeats [(TTTTA)n] in the noncoding region, and of Met/Thr in the coding region of the apolipoprotein(a) [apo(a)] gene. The apo(a) polymorphisms were examined in 184 Korean patients with CAD and 121 healthy subjects. In size polymorphism, there was an inverse association between plasma Lp(a) levels and the apo(a) isoforms in the CAD group (p < 0.005). For (TTTTA)n sequence polymorphism, subjects with the 8/8 genotype were most frequently observed in the two groups. Plasma Lp(a) levels showed a significant difference between the 8/5 versus the 8/8 genotype in the CAD group. On the other hand, Lp(a) levels varied significantly among the genotypes of a Met/Thr polymorphism in normal controls (P < 0.01). The heterozygous genotype had an intermediate level of Lp(a) between the two homozygous genotypes, thereby showing gene dosage effect. Thus, it is possible that the apo(a) polymorphisms are responsible for variations in the Lp(a) levels. PMID- 9856342 TI - Thioltransferase from Arabidopsis thaliana seed: purification to homogeneity and characterization. AB - Thioltransferase is a general GSH-disulfide reductase of importance for redox regulation. The protein thioltransferase has been purified to apparent homogeneity on SDS-PAGE from the Arabidopsis thaliana seed. The purification procedures included DEAE-cellulose ion exchange chromatography, Sephadex G-75 gel filtration, Q-Sepharose ion exchange chromatography, and DEAE-Sephadex A-25 ion exchange chromatography. The enzyme has a molecular mass of 22 kDa and a pI of 4.8, and it is heatstable. The protein had broad specificities for substrates ranging from low-molecular disulfides (S-sulfocysteine and cystine) to protein disulfides (trypsin and insulin). However, it could not reduce the disulfide linkages of ribonuclease A and bovine serum albumin. It could utilize non disulfide substrates such as dehydroascorbic acid and alloxan. The protein can reduce the disulfide bond in 2-hydroxyethyl disulfide with an optimum pH of 8.5. Its activity was greatly activated by monothiol compounds such as reduced glutathione and L-cysteine. PMID- 9856343 TI - Molecular cloning and characterization of a new basic peroxidase cDNA from soybean hypocotyls infected with Phytophthora sojae f.sp. glycines. AB - Differential display techniques were used to isolate cDNA clones corresponding to genes which were expressed in soybean hypocotyls by Phytophthora sojae f.sp. glycines infection. With a partial cDNA clone C20CI4 from the differential display PCR as a probe, a new basic peroxidase cDNA clone, designated GMIPER1, was isolated from a cDNA library of soybean hypocotyls infected with P. sojae f.sp. glycines. Sequence analysis revealed that the peroxidase clone encodes a mature protein of 35,813 Da with a putative signal peptide of 27 amino acids in its N-terminus. The amino acid sequence of the soybean peroxidase GMIPER1 is between 54-75% identical to other plant peroxidases including a soybean seed coat peroxidase. Southern blot analysis indicated that multiple copies of sequences related to GMIPER1 exist in the soybean genome. The mRNAs corresponding to the GMIPER1 cDNA accumulated predominantly in the soybean hypocotyls infected with the incompatible race of P. sojae f.sp. glycines, but were expressed at low levels in the compatible interaction. Soybean GMIPER1 mRNAs were not expressed in hypocotyls, leaves, stems, and roots of soybean seedlings. However, treatments with ethephon, salicylic acid or methyl jasmonate induced the accumulation of the GMIPER1 mRNAs in the different organs of soybean. These results suggest that the GMIPER1 gene encoding a putative pathogen-induced peroxidase may play an important role in induced resistance of soybean to P. sojae f.sp. glycines and in response to various external stresses. PMID- 9856344 TI - Expressed sequence tags of fruits, peels, and carpels and analysis of mRNA expression levels of the tagged cDNAs of fruits from the Fuji apple. AB - In order to understand molecular events during fruit development and provide genetic resources for molecular breeding, 430 expressed sequence tags (ESTs) were generated from randomly selected clones of cDNA libraries prepared from young fruits, peels of mature fruits, and carpels of the Fuji apple (Malus domestica Borkh.). Database comparisons of the ESTs revealed that 180 non-redundant clones showed a high similarity with previously identified genes. Among these, 138 clones exhibited a homology with previously identified plant genes and 12 were identical to genes that were previously identified from apples. The deduced amino acid sequences of 42 clones had a homology to proteins that have not been reported from plants. Eighteen cDNA clones from the young fruit library were selected for studying expression levels and patterns in reproductive organs and leaves. This study revealed that the clones can be classified into 3 different groups based on their expression levels. The first 9 clones were expressed strongly in at least one reproductive organ. Eight of these clones (vacuolar processing protease, sucrose phosphate synthase, arabinogalactan protein, UDP glucose glucosyl transferase, major allergen D1, cystein proteinase inhibitor, lipoxygenase, and protease subunit SUG2) were highly expressed in mature flowers and young fruits, whereas one clone (z-carotene desaturase protein precursor) was preferentially expressed in mature flowers but weakly in young fruits. The second group includes 6 cDNA clones (glucose transport protein, aminomethyl transferase precursor protein, dTDP-D-glucose-4,6-dehydrogenase, 2 types of protein kinase, and selenium binding protein) that were weakly expressed. These clones were characterized by their preferential expression patterns in mature flowers and young fruits. The transcripts of 3 cDNA clones in the third group (vacuolar aminopetidase, beta-galactosidase, and EREBP-4) were detectable only by RT-PCR and they were preferentially expressed in young fruits. These results indicate that most ESTs that were isolated from young fruits are preferentially expressed in reproductive organs and thereby play important roles during reproductive organ development. PMID- 9856345 TI - Cloning and characterization of novel disintegrins from Agkistrodon halys venom. AB - Snake venom disintegrins act as potent inhibitors of platelet aggregation. In this report, we isolated genes encoding novel members of disintegrins through the screening of Agkistrodon halys venom gland cDNA library. Subsequent characterization of positives revealed the presence of distinct disintegrins named salmosinl, 2, and 3, each containing a characteristic RGD/KGD sequence essential for the binding to integrins. Whereas salmosinl was identical to previously described salmosin purified from A. halys venom, salmosin2 and salmosin3 were predicted to be a novel, 73 amino acid protein with a KGD sequence, and an 80 amino acid protein with an additional 7th disulfide bond, respectively. Taken together, this is the first report describing 3 unique disintegrins, namely, salmosinl with RGD, salmosin2 with KGD and salmosin3 with 7 disulfide bonds are found in a single species of venom. Subsequently, to compare the platelet aggregation inhibitory potential of the recombinant protein with that of natural protein, salmosinl was expressed in E. coli and purified to homogeneity. Recombinant and natural salmosin1 inhibited the binding of alphaIIbbeta3 to fibrinogen with an almost identical IC50 value of 2.2 nM and 4.5 nM respectively. Moreover, recombinant salmosinl displayed an IC50 value approximately 5-fold lower than flavoridin, which was previously described as the most potent venom disintegrin so far. In conclusion, we identified 3 disintegrins with distinct properties through the molecular cloning approach and found that the recombinant salmosinl retained one of the most potent alphaIIbbeta3 antagonist activity. PMID- 9856346 TI - Cloning and analysis of the Epstein-Barr virus glycoprotein 350 genes. AB - Membrane glycoprotein 350 (gp350) of the Epstein-Barr virus (EBV) is considered as a major target for vaccine development, since the gp350 has been identified as the virus' mediator for receptor interaction and as an inducer of specific in vitro virus-neutralizing antibodies. In an initial attempt to develop an effective DNA vaccine against an EBV infection, gp350 genes were isolated from SNU-20 and SNU-1103 which are the EBV-infected lymphoblastoid cell lines established in Korea. In addition, the nucleotide sequences of the gp350 genes were determined and compared with those of other EBV strains such as B95-8, P3HR 1/AG876 and M81. Sequence analysis showed that similar high degrees of homology between 2 EBV strains derived from African Burkitt's lymphoma, P3HR-1 and AG876, was shown between the gp350 genes isolated from 2 EBV-infected lymphoblastoid cell lines established in Korea. Furthermore, these 2 Korean and 2 African strains displayed nearly identical patterns of sequence variations from B95-8. In addition, the sequence of the isolated gp350 genes, which have been reported to be associated with the biology of EBV infection, is analyzed. PMID- 9856347 TI - Comparison of the structure and expression pattern for a low molecular weight heat-shock protein cDNA clone from Nicotiana tabacum. AB - We isolated a cDNA clone, named TLHS-1, for a low molecular weight heat-shock protein (LMW HSP) from tobacco. The nucleotide sequence determination of the clone identified an open reading frame for 159 amino acids. To the upstream of the open reading frame, a sequence of 124 nucleotides was determined. To the 3' downstream of the open reading frame, 212 nucleotides were identified which carried a poly(A) tail. A comparison of the open reading frame of TLHS-1 with the previously reported class I LMW HSPs showed a high identity which classified the TLHS-1 as a class I LMW HSP cDNA clone. Reexamination of the amino acid sequences for LMW HSPs including the TLHS-1 argued for the necessity to redraw the consensus region. Six consensus regions were proposed instead of the two consensus regions suggested previously. A RNA blot hybridization for TLHS-1 showed the typical expression pattern of a heat-shock inducible gene from 3 common tobacco cultivars. The genomic DNA blot hybridization for the TLHS-1 gene from the 3 tobacco cultivars showed identical patterns. These results implied that the expression control and structure of the TLHS-1 gene have been well conserved in the tobacco cultivars which are not bred for high temperature stress resistance. PMID- 9856348 TI - No effect of expression of neuron-restrictive silencer factor (NRSF) protein on N type Ca2+ channel alpha 1B gene promoter activity in NS20Y cells. AB - The expression of the N-type voltage-sensitive calcium channel alpha1B gene is restricted to neurons by an 5'-upstream region (-3992 to -1788) containing negative regulation element(s) active in non-neuronal cells (Kim et al., 1997). The neuron-restrictive silencer factor (NRSF) represses the transcription of several neuronal genes in non-neuronal cells by binding to a 21 bp DNA element, termed the neuron-restrictive silencer element (NRSE). To analyze the involvement of NRSF in the neuron-specific expression of the alpha1B gene, the coding region of NRSF cDNA was cloned by PCR and the exogenous NRSF cDNA was transiently cotransfected with tester plasmids into NS20Y neuronal cells which do not contain the endogenous NRSF protein. The luciferase activity of a positive control plasmid NRSEL containing a single copy of the NRSE sequence of the SCG10 gene was repressed 5 fold in HeLa cells containing the endogenous NRSF protein, and its activity was repressed to 44-27% of the control with an increasing amount of exogenous DNA in NS20Y cells. Unlike NRSEL, the promoter activity of the alpha1B subunit-luciferase fusion construct (-3992L) was about 15 fold repressed in HeLa cells compared to NS20Y cells, while any remarkable changes was undetectable in the NRSF expressed NS20Y. These results suggest that the repression of the alpha1B gene in non-neuronal cells may not be mediated by the NRSF function. PMID- 9856349 TI - Isoform-specific interaction of the cytoplasmic domains of Na,K-ATPase. AB - The Na,K-ATPase is a heterodimer consisting of an alpha and a beta subunit, which exchanges intracellular Na+ for extracellular K+ using the energy of ATP hydrolysis. Several studies have demonstrated that the enzyme exists as an (alphabeta)2 heterotetramer, an oligomer of alphabeta dimers within the cell membrane, at least during some portion of the transport cycle although its functional significance is unknown. In our study, we employed the yeast two hybrid system to identify the cytoplasmic domains of the Na,K-ATPase which might be involved in intersubunit and/or intrasubunit interactions to form higher order oligmers. Our data demonstrate that the N-terminus and the cytoplasmic loop 1 of the alpha2 subunit interact with each other, while those of the alpha1 subunit do not, suggesting that the interaction is isoform-specific. Therefore, the N terminal and the cytoplasmic loop 1 might be the regions where the alpha2 subunit, which are involved in alpha alpha interactions, stabilize Na,K-ATPase as alphabeta protomer, diprotomer, or higher order oligomer because the interaction can be intrasubunit as well as intersubunit interactions. Our study suggests that there may be an isoform-specific difference in the alpha-alpha interaction and that the isoform-specific interaction may contribute significantly to the differences of the physiological function and regulation among the alpha isoforms. PMID- 9856350 TI - CRP.cAMP-dependent transcription activation of the Escherichia coli pts Po promoter by the heat shock RNA polymerase (Esigma32) in vitro. AB - The P0 promoter of the Escherichia coli pts operon that is activated during growth in the presence of glucose contains a recognition sequence for the RNA polymerase holoenzyme containing sigma 70 (Esigma70) which is the RNA polymerase holoenzyme responsible for the transcription of heat shock genes, and a putative recognition element for Esigma32, the Esigma32 was found to be able to transcribe from the pts P0 promoter with in vitro transcription assays using purified RNA polymerase holoenzymes. Esigma32 and Esigma70 used the same transcriptional start site and both RNA polymerases were activated in the presence of a cAMP receptor protein (CRP) and cAMP complex (CRP.cAMP). These results suggest the possibility that the contact between CRP.cAMP and the alpha subunit of the RNA polymerase can be made regardless of the kind of sigma subunit that is associated with the core RNA polymerase. PMID- 9856351 TI - Circulating soluble CD5 in atopic dermatitis. AB - The CD5/Leu-1 is involved in the activation of the T-cell helper function through T/B-cell collaboration by CD5/CD72 interaction. T-cell function is known to be dysregulated in atopic dermatitis. However, to date, the role of CD5 has not been investigated in atopic dermatitis, nor has the presence of circulating soluble CD5 been reported in atopic dermatitis. Five patients with atopic dermatitis who showed typical symptoms, 5 acute febrile patients and 5 normal subjects were tested. Peripheral blood mononuclear cells and plasma were separated. The T- and B-cells were separated using immunomagnetic beads. Reverse transcription polymerase chain reaction was performed using CD5 specific primers. Immunoblotting with the mouse antiCD5 monoclonal antibody was done. Circulating soluble CD5 was present only in 4 out of 5 atopic patients. However, it was not detected in acute febrile patients nor in normal subjects. CD5 mRNA expression was detected in all atopic patients and acute febrile patients. CD5 mRNA expression in T- and B-cells was tested in patients with atopic dermatitis and was detected only in the T-cells. In this study, circulating soluble CD5 was detected in atopic patients and soluble CD5 was suspected to participate in the pathogenesis of atopic dermatitis. CD5 mRNA expression was detected only in T cells, which suggests that circulating soluble CD5 might be produced from T cells. PMID- 9856352 TI - A site in the dinucleotide-fold domain contributes to the accuracy of tRNA selection by Escherichia coli methionyl-tRNA synthetase. AB - Interactions of specific amino acid residues of the carboxyl-terminal domain of MetRS with the CAU anticodon of tRNAMet assure accurate and efficient aminoacylation. The substitution of one such residue, Trp461 by Phe, impairs the binding of cognate tRNA, but enhances the binding of noncognate tRNAs, particularly those containing G at the wobble position. However, the enhanced binding of noncognate tRNAs is not accompanied by the increased aminoacylation of these tRNAs. A genetic screening procedure was designed to isolate methionyl-tRNA synthetase mutants which were able to aminoacylate a GGU (threonine) anticodon derivative of tRNAfMet. One such mutant, obtained from W461F MetRS, had an Ile29 to Thr substitution in helix A located in the amino-terminal dinucleotide-fold domain that forms the site for amino acid activation. Analysis of the catalytic properties of the I29T/W461F enzyme indicates that the mutation in helix A of the dinucleotide-fold domain affects kcat for aminoacylation of tRNAs having a GGU threonine anticodon. Interactions with cognate tRNAfMet (CAU), as well as with methionine and ATP were not affected by the Ile29 to Thr substitution. We conclude that the I29T substitution leads to a slight adjustment of the alignment of the CCA stem of noncognate tRNAs (GGU) in the catalytic domain of the enzyme, reflected in the increase in kcat, which also allows mischarging in vivo. A function of Ile29 is therefore to minimize the mischarging of tRNAThr (GGU) by methionyl-tRNA synthetase. The methods described here provide useful tools for examining the mechanisms of tRNA selection by aminoacyl-tRNA synthetases. PMID- 9856353 TI - Augmentation of therapeutic antitumor immunity by B16F10 melanoma cells transfected by interferon-gamma and allogeneic MHC class I cDNAs. AB - The cDNAs for interferon-gamma (IFN-gamma) and allogeneic H-2Kd molecules were transfected into highly metastatic B16F10 melanoma cells (H-2b), and the synergistic effects of the antitumor immune responses by the doubly transfected cells (B16/Kd/IFN-gamma cells) were investigated in C57BL/6 mice (H-2b). The singly transfected B16F10 cells with either IFN-gamma or H-2Kd cDNA (B16/IFN gamma or B16/Kd cells) were used as controls. The B16/Kd/IFN-gamma cells secreted biologically active IFN-gamma, and strongly expressed both syngeneic and allogeneic MHC class I antigens (H-2Kb and H-2Kd) on the same cell construct. Immunization with the doubly transfected B16/Kd/IFN-gamma cells induced higher anti B16F10 cellular cytotoxic responses than the single transfected B16/IFN gamma or B16/Kd cells. Lyt-2.2 (CD8)+ T-cells were a major effector cell-type involved in the anti B16F10 responses and their cytotoxic activities were augmented in the immunized mice with the B16/Kd/IFN-gamma cells, as demonstrated by in vitro depletion experiments. The survival period of melanoma-bearing mice treated with the B16/Kd/IFN-gamma cells was significantly longer than that treated with the B16/IFN-gamma or B16/Kd cells. Furthermore, the treatment with the B16/Kd/IFN-gamma cells was capable of greatly inhibiting lung metastasis from small, established B16F10 footpad tumors. These results suggest that the augmented immunotherapeutic potentials can be achieved by the vaccination with IFN-gamma and allogeneic MHC class I genes transfected B16F10 melanoma cells. PMID- 9856354 TI - Programmed changes of cell cycle regulators by serum deprivation regardless of skeletal myocyte differentiation. AB - Permanent withdrawal of skeletal myoblasts from the cell cycle precedes differentiation. We examined the changes of protein levels of cell cycle regulators and the activities of CDKs in differentiating (by serum deprivation) or in differentiation-inhibited (by serum deprivation + TGF-beta1 treated) C2C12 skeletal myocytes. Regardless of differentiation, protein levels of most cyclins declined over time while this effect was delayed slightly by TGF-beta1 for cyclins D1 and F. Although the protein levels of CDKs gradually decreased over time, the activities of CDK2 and cdc2 decreased dramatically between 0 and 12 h after serum deprivation in both groups. Decreased CDK2 and cdc2 activities were temporally related to decreased DNA synthesis. While the p27KIP1 protein increased in both groups, the p21CIP1 protein increased only in differentiating cells. In vivo, protein expressions of cyclins, CDKs, and p21CIP1 were high in fetal, but almost undetectable in adult skeletal muscle. In contrast, the levels of p27KIP1 protein in skeletal muscle were high throughout development. Thus, changes of cell cycle regulators in differentiating C2C12 myocytes paralleled those during skeletal muscle development of mice. These results suggest that the rapid reduction of DNA synthesis and activities of CDKs slow reduction of protein levels of cyclins and CDKs, and that the increase of p27KIP1 are programmed mechanisms upon mitogen deprivation regardless of differentiation in skeletal myocytes. PMID- 9856355 TI - Angiogenesis and hypertension. AB - BACKGROUND: The formation of new blood vessels is an important process in embryonic development and in physiological repair processes. Abnormalities in blood vessel growth have been associated with various pathologies. HYPERTENSION AND IMPAIRED VASCULAR GROWTH: The basic observation underlying the hypothesis that essential hypertension is based on an impaired capacity for vascular growth is the nature of the structural alterations of microvascular beds in essential hypertension. Recent advances in understanding the molecular and cellular mechanisms of vascular growth suggest that the remodeling of individual vessels and vascular networks in hypertension may be a pathological variant of the formation of mature networks. PATHOGENESIS OF IMPAIRED VASCULAR GROWTH: Genetic and fetal influences appear to have significant effects in determining impaired vascular development as an early cause of essential hypertension. PMID- 9856356 TI - Hypertension in master endurance athletes. AB - OBJECTIVE: To determine whether long-term very vigorous endurance training prevents hypertension. DESIGN: Cohort study of master orienteering runners and controls. SETTING: Finland. SUBJECTS AND METHODS: In 1995, a health questionnaire was completed by 264 male orienteering runners (response rate 90.4%) who had been top-ranked in competitions among men aged 35-59 years in 1984, and by 388 similarly aged male controls (response rate 87.1%) who were healthy at the age of 20 years and free of overt ischemic heart disease in 1985. MAIN OUTCOME MEASURE: Self-report of medication for hypertension. RESULTS: In the endurance athlete group, the crude prevalence (8.7%) of subjects who had used medication for hypertension was less than a third of that in the control group (27.8%). Even after adjusting for age and body mass index, the difference between the groups was still significant (odds ratio for athletes 0.43, 95% confidence interval 0.25 0.76). CONCLUSIONS: Long-term vigorous endurance training is associated with a low prevalence of hypertension. Some of the effect can be explained by a lower body mass, but exercise seems to induce a lower rate of hypertension by other mechanisms than by decreasing body weight PMID- 9856357 TI - Plasma fibrinogen and incident hypertension in the Atherosclerosis Risk in Communities (ARIC) Study. AB - BACKGROUND: Several cross-sectional studies have reported a positive association between plasma fibrinogen levels and prevalent hypertension. Other studies have reported a positive association between hypertension and whole-blood or plasma viscosity, to which fibrinogen contributes. To our knowledge, there has been no prospective study of fibrinogen and incident hypertension. SUBJECTS AND METHODS: We measured plasma fibrinogen levels in a population-based cohort study of middle aged adults and related it to the occurrence of incident hypertension (systolic blood pressure > or = 140 mmHg or diastolic blood pressure > or = 90 mmHg or use of antihypertensive medication) over 6 years. RESULTS: There was a moderately strong positive association between fibrinogen levels and prevalent hypertension in both men and women, with the odds of hypertension elevated by 50% for the highest fibrinogen quartile versus the lowest. Among 7884 participants at risk, 1609 developed hypertension over 6 years. Adjusted for age, race, field center and baseline systolic blood pressure, the odds ratio of incident hypertension in relation to fibrinogen quartiles was 1.0, 1.07, 1.21 and 1.43 in men (P= 0.003 for trend) and 1.0, 0.92, 0.99 and 0.99 in women (P= 0.89 for trend). After adjustment for other risk factors, the odds ratios were 1.0, 1.03, 1.15 and 1.29 (P= 0.045 for trend) in men and remained nonsignificant in women. CONCLUSIONS: Despite a moderately strong positive association between fibrinogen levels and prevalent hypertension in both sexes, there was only a weak positive association between fibrinogen levels and incident hypertension in men and no association in women. Whether an elevated fibrinogen level is a risk factor for, or a consequence of, hypertension remains unclear. PMID- 9856358 TI - Seasonal variations in home and ambulatory blood pressure in the PAMELA population. Pressione Arteriose Monitorate E Loro Associazioni. AB - OBJECTIVE: Clinic blood pressure values are known to change according to seasonal influences. We therefore examined home and 24 h ambulatory blood pressure values to determine whether these measurements are also affected by the seasons. DESIGN AND METHODS: In 2051 subjects of the Pressione Arteriose Monitorate E Loro Associazioni (PAMELA) study, we measured clinic (sphygmomanometric measurements), home (semi-automatic device) and ambulatory (Spacelabs 90207) systolic blood pressure, diastolic blood pressure and heart rate. Because the overall sample was evenly distributed over each month (except August), we were able to make a cross sectional determination of whether the values differed between seasons. The corresponding heart rates were also evaluated. RESULTS: As expected, summer was associated with the lowest clinic blood pressure and winter with the highest, and this was the case also for home and 24 h average blood pressure, although seasonal differences in the latter were less pronounced. Seasonal clinic, home and ambulatory blood pressure patterns were similar for normotensive subjects (n = 1152), untreated hypertensives (n = 540) and treated hypertensives (n = 359). Heart rate values did not differ by season. CONCLUSIONS: Seasonal influences on blood pressure are not limited to conventional measurements but characterize daily values as well. These effects are visible in both normal and elevated blood pressure values, regardless of the effect of antihypertensive drugs. This has implications both for the clinician and for studies aimed at evaluating the effects of antihypertensive treatment. PMID- 9856359 TI - Intima-media thickness and diameter of carotid and femoral arteries in children, adolescents and adults from the Stanislas cohort: effect of age, sex, anthropometry and blood pressure. AB - OBJECTIVES: To study carotid and femoral intima-media thicknesses and diameters in relation to age, sex, morphologic status and blood pressure. PARTICIPANTS: The subjects were 369 men and women (aged 10-54 years) from the Stanislas cohort, with no known cardiovascular disease. METHODS: Intima-media thicknesses and diameters were measured by B-mode ultrasonography. The effects of sex, age, smoking, anthropometric variables, cholesterol and blood pressure were studied using bivariate and regression analysis. RESULTS: Carotid and femoral intima media thicknesses were not affected by age nor by sex up to 18 years of age. Thereafter, they increased sharply in men and remained higher than in women. Values were correlated with systolic blood pressure only in men, and with fat free mass in children and young adults only at the femoral site. Smoking, body mass index and fat mass were associated with intima-media thicknesses only in adults. Carotid diameter was little affected by age during childhood and in adults. Femoral diameter increased up to the age of 18 in both sexes and remained unaffected by age thereafter. This increase was more pronounced in boys, and so values became consistently greater in males aged over 14 years. Carotid diameter was correlated with body mass index or fat mass whereas femoral diameter was correlated with weight or fat-free-mass in children and men. The opposite was observed in women. CONCLUSION: Sex differences occur before adolescence for arterial diameter, but only at an adult age for intima-media thickness. In young subjects, carotid geometry seems to be influenced by blood pressure and excess body weight, while femoral artery geometry seems to be related to blood pressure and body growth. PMID- 9856360 TI - Angiotensin converting enzyme inhibition reduces the expression of transforming growth factor-beta1 and type IV collagen in diabetic vasculopathy. AB - OBJECTIVE: The purpose of this study was to assess the role of transforming growth factor (TGF)-beta1 in the development of diabetes-associated mesenteric vascular hypertrophy and in the antitrophic effect of angiotensin converting enzyme inhibitors. DESIGN AND METHODS: Streptozotocin-induced diabetic and control Sprague-Dawley rats were randomly allocated to treatment with the angiotensin converting enzyme inhibitor ramipril or to no treatment and were killed 1 or 3 weeks after the streptozotocin injection. Blood was collected and mesenteric vessels removed. Mesenteric vascular weight was measured and TGF-beta1 and alpha1 (type IV) collagen messenger (m)RNA levels were analysed by Northern analysis. Immunohistochemical analyses for TGF-beta1 and type IV collagen were also performed. RESULTS: The diabetic rats had increased mesenteric vessel weight at 3 weeks but not at 1 week and a concomitant rise in mesenteric TGF-beta1 and in alpha1 (type IV) collagen mRNA levels. Ramipril treatment attenuated mesenteric vessel hypertrophy and prevented the increase in TGF-beta1 and alpha1 (type IV) collagen mRNA levels after 3 weeks of diabetes. The immunohistochemical analysis revealed that diabetes was associated with increased TGF-beta1 and type IV collagen protein and extracellular matrix accumulation in mesenteric vessels, and this increase was reduced by ramipril treatment. CONCLUSIONS: These results support the concept that TGF-beta is involved in the changes associated with diabetic vascular disease, and suggest a mechanism by which angiotensin converting enzyme inhibitors exert their antitrophic effects. PMID- 9856362 TI - Shear stress abnormalities contribute to endothelial dysfunction in hypertension but not in type II diabetes. AB - BACKGROUND: The relative contribution of the various hemodynamic and metabolic mechanisms leading to endothelial dysfunction may be different in specific vascular diseases. Since shear stress is one of the main mechanical stimuli of endothelial cells, the aim of this study was to investigate its contribution to endothelial dysfunction in two distinct vascular diseases, hypertension and type II diabetes. SUBJECTS AND METHODS: We measured the radial artery diameter at baseline, after ischemic vasodilation and after nitroglycerin vasodilation in 16 untreated patients with high blood pressure, in 15 type II normotensive diabetic patients and in 17 healthy controls. Wall shear stress was evaluated by simultaneous measurements of whole blood viscosity and blood flow velocity. RESULTS: In diabetic patients, whole blood viscosity was significantly higher whereas wall shear stress was similar compared to controls. In hypertensive patients, whole blood viscosity was higher and wall shear stress was lower than in controls. Endothelium-dependent vasodilation was impaired in both hypertensive and diabetic patients (P < 0.01) after adjustment for age, sex, body mass index and postnitroglycerin vasodilation. When adjustments were made for maximal systolic shear stress, endothelium-dependent vasodilation remained lower in the diabetic patients (P < 0.01), but not in those with high blood pressure compared to controls. CONCLUSIONS: In hypertension, endothelium-dependent vasodilation is mainly due to a chronic decrease in shear stress (the most important physiological stimulus of the endothelial cells) with no major intrinsic endothelial cell dysfunction. In contrast, in diabetics, the lower endothelium dependent vasodilation was not the result of an altered shear stress. PMID- 9856361 TI - Role of nitric oxide in vascular hyper-responsiveness to norepinephrine in hypertensive Dahl rats. AB - OBJECTIVE: To determine whether the abnormal vascular responses observed in salt sensitive hypertension are caused by an impairment in vascular nitric oxide function. DESIGN: Isometric tension was measured in aortic rings isolated from Dahl salt-sensitive and salt-resistant rats fed a regular-salt (0.4% NaCl) or a high-salt (8% NaCl) diet, with and without inhibition of endogenous nitric oxide synthesis. METHODS AND RESULTS: Systolic arterial pressure, measured weekly by the tail-cuff method, increased markedly in DS rats with a high-salt diet but did not increase in the other groups. In aortic rings, norepinephrine evoked dose dependent contractions which were significantly increased in rings from DS rats with a high-salt diet Pretreatment with Nomega-nitro-L-arginine methyl ester (L NAME), a nitric oxide synthase inhibitor, increased the norepinephrine-induced contraction in all groups and abolished differences in contractile responses between high-salt DS rats and the other groups. Acetylcholine induced endothelium dependent relaxation, which was significantly depressed in high-salt DS rats. L NAME attenuated the acetylcholine-induced relaxation in all groups and abolished the difference in relaxation response between high-salt DS rats and the other groups. Sodium nitroprusside-induced relaxation was significantly depressed in high-salt DS rats. CONCLUSIONS: Vascular hypercontractile responses to norepinephrine in DS hypertensive rats can, in part, be explained by an impairment in endothelial nitric oxide production. PMID- 9856363 TI - Structural analysis and evaluation of the 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) gene in human essential hypertension. AB - AIM: Mutations of the 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) gene cause the syndrome of apparent mineralocorticoid excess, a rare autosomal recessive form of hypertension. We therefore investigated the question of whether variants of the 11beta-HSD2 gene can contribute to genetic susceptibility to essential hypertension. SUBJECTS AND METHODS: We performed a linkage study in 162 French hypertensive sibships using the affected sib-pair method on 347 sibling pairs and a polymorphic microsatellite marker that we identified in a 30 kb cosmid clone containing the 11beta-HSD2 gene. The coding sequence, introns 2-4 and 350 bp of the 5'-flanking region of the 11beta-HSD2 gene were screened for polymorphisms by polymerase chain reaction/single-strand conformation polymorphism, and a single polymorphism, Glu178/Glu (G534A), was identified in exon 3, which did not change the encoded amino acid sequence. A case-control study was conducted on 370 hypertensive subjects with a positive family history of hypertension and 783 French subjects with hypertension with or without a family history of hypertension, compared with 313 normotensive control subjects, all of whom were analyzed for the newly identified bi-allelic polymorphism. RESULTS: Statistical analyses using the affected sib-pair method did not show significant linkage between the 11beta-HSD2 microsatellite marker and hypertension. Furthermore, no positive association with hypertension was found with the Glu178/Glu (G534A) polymorphism. CONCLUSION: Our data do not suggest that variants of the 11beta-HSD2 gene contribute substantially to essential hypertension in Caucasians. PMID- 9856364 TI - Heart rate as marker of sympathetic activity. AB - OBJECTIVE: To determine the value of the supine heart rate as a marker of sympathetic tone by assessing, in a large group of subjects, the relationships between this parameter and two other indices of sympathetic activity, plasma norepinephrine and sympathetic nerve traffic. PATIENTS AND METHODS: We studied 243 subjects aged 50.0+/-12.1 years (mean +/- SD). Of these, 38 were normotensive healthy controls, 113 subjects had untreated essential hypertension, 27 were obese normotensives and 65 had congestive heart failure. In each subject, over a 10 min supine period, we measured mean arterial pressure (Finapres), heart rate (electrocardiogram), venous plasma norepinephrine (high-performance liquid chromatography) and efferent postganglionic muscle sympathetic nerve activity (microneurography at a peroneal nerve). RESULTS: In the whole study group, supine heart rate was correlated with both plasma norepinephrine (r = 0.32, P < 0.0001) and muscle sympathetic nerve activity (r = 0.38, P < 0.0001). This was also the case in the normotensive obese subjects and the heart failure subjects considered separately. Heart rate values were greater in the obese and the heart failure patients than in controls (75.1+/-13.0 and 78.2+/-13.0 versus 69.2+/-11.6 beats/min; P < 0.05 and P < 0.001, respectively), as were plasma norepinephrine (362.7+/-202 and 400.3+/-217 versus 230.4+/-126 pg/ml; P < 0.01 and P < 0.001, respectively) and muscle sympathetic nerve activity (44.1+/-14.7 and 55.3+/-14.3 versus 27.8+/-11.0 bursts/min; P < 0.001 for both). In contrast, in the essential hypertensive subjects, no significant relationship was found between these three indices of sympathetic activity. Furthermore, in the hypertensives, the heart rate was not increased, at variance with the sympathetic nerve traffic, which was greater than in controls (36.2+/-10.0 versus 27.8+/-11.0 bursts/min, P < 0.001). CONCLUSIONS: These data suggest that the supine heart rate can be regarded as a marker of intersubject differences in sympathetic tone, and that this is the case both in the general population and in those with cardiovascular diseases. Its value for this purpose is limited, however, and the limitations may be more evident in essential hypertension than in conditions such as obesity and heart failure. PMID- 9856365 TI - Power spectral analysis of the heart rate in hypertensive patients with and without left ventricular hypertrophy: the effect of a left ventricular mass reduction. AB - OBJECTIVE: The aim of this study was to evaluate the spectral analysis of the heart rate in normotensive subjects and in hypertensive patients with and without left ventricular hypertrophy (LVH), under basal conditions and after a reduction in left ventricular mass. SUBJECTS AND METHODS: In 12 normotensive subjects and 22 hypertensive patients (14 with and eight without LVH), we performed 24 h electrocardiogram Holter monitoring, ambulatory blood pressure monitoring and an echocardiographic study. Sequences of 512 R-R intervals, during daytime, afternoon and night-time periods, were taken for an evaluation of spectral analysis (Box-Jenkins method). We then calculated the absolute and percentage power spectral density of the peak centred at 0.10 Hz (low-frequency peak) and at 0.25 Hz (high-frequency peak). RESULTS: At baseline, a daytime to night-time decrease in the low-frequency peak was detected in normotensives (P < 0.01) and in hypertensives without LVH (P < 0.01), while no change was observed in hypertensives with LVH. The power spectral density low-frequency peak during the daytime and night-time was significantly greater in hypertensives with LVH than in those without LVH (P < 0.001) and in normotensive subjects (P < 0.001). Fourteen of these patients with LVH were given effective long-term antihypertensive treatment and were studied again 20 days after the treatment had been withdrawn, when blood pressure had increased to pretreatment values. In eight patients showing a reduction in LVH, we found a significant decrease in the power spectral density low-frequency peak and an increase in the high-frequency peak during daytime and night-time in respect to basal conditions, and circadian variations in the spectral indices of heart rate variability were restored. In contrast, in six patients without reversal of LVH, the power spectral density low frequency peak did not change in respect to basal conditions and remained significantly higher in comparison with the patients with LVH regression. CONCLUSION: A reduction in LVH may be associated with restoration of daytime to night-time cardiac autonomic control, as evaluated by a power spectral analysis of the heart rate. PMID- 9856366 TI - Captopril does not improve insulin action in essential hypertension: a double blind placebo-controlled study. AB - OBJECTIVE: To compare the effect of captopril with that of placebo on peripheral and hepatic insulin action in essential hypertension, in light of evidence that insulin resistance is associated with cardiovascular risk. DESIGN: Randomized, double-blind, placebo-controlled, crossover trial, with 8 week treatment periods of captopril and placebo preceded and separated by 6 weeks of placebo. SETTING: Belfast teaching hospital. PATIENTS: Eighteen Caucasian nondiabetic patients (10 males), aged under 65 years, with essential hypertension, recruited from general practices in the greater Belfast area. INTERVENTIONS: Captopril at 50 mg twice a day or placebo twice a day for two 8 week treatment periods. MAIN OUTCOME MEASURES: Peripheral and hepatic insulin sensitivity assessed by glucose clamps. RESULTS: Fourteen patients completed the study. Mean (+/- SEM) levels of fasting glucose, fasting insulin and postabsorptive hepatic glucose production were similar after captopril and placebo (5.4+/-0.1 versus 5.4+/-0.1 mmol/l, 10.6+/ 2.2 versus 9.5+/-1.1 mU/l, 11.2+/-0.6 versus 11.0+/-0.5 mmol/kg per min, respectively). During hyperinsulinaemia, hepatic glucose production was suppressed to comparable levels after both treatments (4.8+/-0.6 versus 4.3+/-0.6 mmol/kg per min) and exogenous glucose infusion rates required to maintain euglycaemia were also similar (30.0+/-2.6 versus 30.3+/-2.6 mmol/kg per min). CONCLUSION: Captopril therapy in uncomplicated essential hypertension has no effect on peripheral or hepatic insulin sensitivity. PMID- 9856367 TI - Increased oxidative stress in renal proximal tubules of the spontaneously hypertensive rat: a mechanism for defective dopamine D1A receptor/G-protein coupling. AB - AIM: Defective dopamine D1A dopamine receptor/G-protein coupling has been demonstrated in renal proximal tubules of the spontaneously hypertensive rat (SHR). In the present study, we aimed to analyze the underlying mechanisms through which such defects are introduced into the D1A receptor protein of SHR. MATERIALS AND METHODS: The oxidative state of SHR proximal tubules was analyzed by measuring lipid peroxidation. D1A receptor/G-protein coupling was measured following the induction of oxidative stress in normotensive Wistar-Kyoto (WKY) rats. RESULTS: For the first time, an increased state of oxidative stress was demonstrated in SHR proximal tubules compared with those of normotensive controls, WKY and Sprague-Dawley rats. Lipid peroxidation levels in SHR were significantly higher by 66 and 79%, relative to WKY or Sprague-Dawley rats, respectively. Hydrogen peroxide treatment of proximal tubules from SHR, WKY and Sprague-Dawley rats induced an additional increase in lipid peroxidation in a dose-dependent manner, although the percentage induction was lower in SHR than in WKY and Sprague-Dawley rats. This induction of lipid peroxidation in WKY rats resulted in a loss of D1A/G-protein coupling, with no decrease in receptor protein. Treatment of WKY rat proximal tubules with an antioxidant, ascorbic acid, or a reducing agent, dithiothreitol, induced D1A receptor/G-protein coupling. CONCLUSIONS: These data indicate that D1A receptor/G-protein coupling is modulated by changes in redox states. Therefore, the D1A receptor/G-protein coupling in SHR may have been damaged by reactive oxygen species released as a result of the elevated oxidative stress seen in the proximal tubules. PMID- 9856368 TI - The Verapamil in Hypertension and Atherosclerosis Study (VHAS): results of long term randomized treatment with either verapamil or chlorthalidone on carotid intima-media thickness. AB - BACKGROUND: It is unclear whether the carotid intima-media thickness can be influenced by antihypertensive treatment and whether some antihypertensive agents, such as calcium antagonists, may have a greater effect on this parameter than others, such as diuretics. The present paper reports the principal results of the ultrasound substudy of the randomized, prospective, controlled, Verapamil in Hypertension and Atherosclerosis Study (VHAS). DESIGN AND METHODS: In 498 hypertensive patients in eight Italian centres, randomized to either verapamil (240 mg once a day) or chlorthalidone (25 mg once a day), a B-mode ultrasound scan was performed according to a standardized procedure at baseline and after 3, 12, 24, 36 and 48 months of treatment. The maximum intima-media thicknesses of the far walls of common, bifurcation and internal carotid arteries were measured bilaterally, and the following indices calculated: the mean thickness at the six measured sites, the mean thickness at the common and bifurcation sites and the single maximum thickness. The primary endpoint for treatment efficacy was the slope of the change over 4 years (rate of change, mm/year), corrected by using the initial mean over the six sites (baseline + 3 months) as a covariate (mm/year per mm). The patients were also classified into three strata according to their baseline single maximum thickness: those with normal carotid arteries (single maximum ( 1 mm), those with thickened carotid arteries (single maximum > 1 and < or = 1.5 mm and those with carotid plaques (single maximum > 1.5 mm). RESULTS: Among the 456 patients with satisfactory baseline ultrasound readings, 33% were classified with normal carotid arteries, 27% with thickened carotid arteries and 40% with plaques. In the intention-to-treat population (377 patients with ultrasound measurements taken on at least three different occasions over a period of at least 2 years), the rate of change in the mean thickness at the six sites measured was rather small (0.015 mm/year), but significantly (P < 0.05) smaller in patients with plaques (0.003 mm/year) than in patients with thickened or with normal carotids (0.023 and 0.025 mm/year, respectively). When related to initial values, the rate of change in the mean thickness at the six sites had a negative slope (-0.059 mm/year per mm, P < 0.01). Although rates of change in the carotid intima-media thickness in unstratified patients were not different in those treated with verapamil or with chlorthalidone, when changes in the mean thickness of six sites were related to the initial value, the slope of this relationship was significantly different in the two treatment groups (verapamil -0.082 versus chlorthalidone -0.037 mm/year per mm, P < 0.02). The blood pressure-lowering effect of the two randomized treatments was similar. Taking fatal and nonfatal, major and minor cardiovascular events together, there were 19 events in the verapamil group and 35 in the chlorthalidone group, with a significantly (P < 0.01) greater incidence in patients with plaques, and among patients with plaques in those who were randomized to chlorthalidone (P < 0.05). CONCLUSIONS: In accord with evidence from animal models of atherosclerosis, the calcium antagonist verapamil was more effective than the diuretic chlorthalidone in promoting regression of thicker carotid lesions. Changes in the carotid intima-media thickness were small in both groups, and the differences between the changes under the two treatments were consequently small, but the observation that these small differences in carotid wall changes were paralleled by differences in the incidence of cardiovascular events (better intima-media thickness regression with verapamil paralleled by a lower cardiovascular event rate) suggests that even small effects on carotid plaques may have clinical and prognostic relevance. PMID- 9856369 TI - Low-dose antihypertensive therapy with 1.5 mg sustained-release indapamide: results of randomised double-blind controlled studies. European study group. AB - OBJECTIVE: In accordance with international recommendations on the need to decrease doses of antihypertensive drugs, a low-dose (1.5 mg) sustained-release (SR) formulation of indapamide was developed to optimize the drug's efficacy : safety ratio. The aim of this work was to evaluate the benefit of a low-dose diuretic by consolidating the efficacy and safety results of two clinical trials with a similar design. PATIENTS AND METHODS: Clinical data were obtained in two European randomized double-blind studies with 690 mild to moderate hypertensive patients (95 mmHg < or = supine diastolic blood pressure < or = 114 mmHg using a mercury sphygmomanometer) treated respectively for 2 and 3 months, with a mean age of 53 and 57 years, 44 and 57% males, mean supine diastolic blood pressure of 100.6 and 102.5 mmHg and mean supine systolic blood pressure of 161.0 and 164.5 mmHg. RESULTS: The first study, a dose-finding study with indapamide SR at 1.5, 2 and 2.5 mg versus placebo and the immediate-release (IR) formulation of indapamide, showed that the 1.5 mg dosage of the new indapamide formulation had an improved antihypertensive efficacy : safety ratio. The second study confirmed the equivalence of blood pressure reductions with 1.5 mg indapamide SR and 2.5 mg indapamide IR, and better acceptability with 1.5 mg indapamide SR, particularly in the number of patients with serum potassium levels < 3.4 mmol/l, which was reduced by more than 50%. The long-term efficacy of 1.5 mg indapamide SR was observed through a 9-month open-treatment follow-up to the second study. CONCLUSION: The 1.5 mg SR formulation of indapamide has an improved antihypertensive efficacy : safety ratio, which is in accordance with international recommendations for the use of low-dose antihypertensive drugs and diuretics in first-line therapy of hypertension. PMID- 9856370 TI - The smoothness index: a new, reproducible and clinically relevant measure of the homogeneity of the blood pressure reduction with treatment for hypertension. AB - OBJECTIVE: To introduce a new method, the smoothness index, for assessing the homogeneity of 24 h blood pressure reduction by antihypertensive treatment and to compare it with the trough : peak ratio; and to assess the ability of both indices to predict a reduction in the left ventricular mass index induced by treatment. PATIENTS AND METHODS: In 174 patients with essential hypertension and left ventricular hypertrophy, enrolled in the Study on Ambulatory Monitoring of Pressure and Lisinopril Evaluation (SAMPLE), aged 20-65 years, we measured clinic blood pressure, 24 h ambulatory blood pressure and the left ventricular mass index (echocardiography) before and after treatment with lisinopril at 20 mg with the addition of 12.5 or 25 mg hydrochlorothiazide as needed to reach a sufficient blood pressure reduction. The following parameters were computed for systolic and diastolic ambulatory blood pressure: (1) hourly and 24 h blood pressure averages (+/- SD) at baseline and after 3 and 12 months of treatment; (2) hourly blood pressure changes from baseline after 3 and 12 months of treatment, and their average (+/- SD) over 24 h; (3) the trough : peak ratio after 3 and 12 months of treatment; and (4) the smoothness index after 3 and 12 months of treatment Similar calculations were also performed at the end of a final study month during which active treatment was withdrawn and placebo was substituted (n = 164). RESULTS: The smoothness index for systolic and diastolic ambulatory blood pressure computed after 3 months of treatment was more closely related to its corresponding values after 12 months of treatment than the trough : peak ratio values computed after the same time periods were (r = 0.68 versus 0.38 for systolic and 0.68 versus 0.42 for diastolic blood pressure, respectively). The smoothness index showed an inverse correlation with the 24 h standard deviation of systolic and diastolic blood pressure (r = -0.25 and -0.16, P < 0.01 and < 0.05, respectively, for 12 months of treatment), while the trough : peak ratio did not (r = -0.01 to -0.12, NS). A treatment-induced reduction in the left ventricular mass index was related to the smoothness index for systolic and diastolic blood pressure (r = -0.35 and -0.32, P< 0.001 with 12 months of treatment), but not to the corresponding trough : peak ratios. CONCLUSIONS: The smoothness index identifies the occurrence of a balanced 24 h blood pressure reduction with treatment and correlates with the favourable effects of treatment on left ventricular hypertrophy better than the commonly used trough : peak ratio. PMID- 9856371 TI - Calcium supplementation in patients with essential hypertension: assessment by office, home and ambulatory blood pressure. AB - OBJECTIVE: To study the effects of a high calcium intake in hypertensive patients by blood pressure monitoring. DESIGN: In a randomized crossover study, patients were assigned to an 8-week calcium supplementation period and an 8-week control period. The subjects were given 25 mmol/day (1 g/day) of calcium as calcium carbonate during the intervention period. SETTING: A hypertension clinic in a tertiary teaching hospital. PATIENTS: Sixty untreated or treated hypertensive patients (35 men and 25 women, mean age 58 years) with office systolic/diastolic blood pressure > or = 140/90 mmHg. MAIN OUTCOME MEASURES: Office blood pressure, home blood pressure (last 7 days), and ambulatory 24 h blood pressure (every 30 min using TM-2421). RESULTS: The serum calcium concentration and urinary calcium excretion increased significantly with calcium supplementation. Office, home and 24 h blood pressure were lower in the calcium period than in the control period, although the differences were small (mean +/- SEM office blood pressure: 1.2+/ 1.2/1.1+/-0.7 mmHg; home blood pressure: 1.9+/-0.7/1.3+/-0.6 mmHg; 24 h blood pressure: 1.2+/-0.8/0.9+/-0.5 mmHg,), and significant only for home systolic and diastolic blood pressures. The difference in home systolic blood pressure was inversely correlated with the level of home blood pressure in the control period and with the difference in urinary calcium. The difference in 24 h systolic blood pressure was positively correlated with the control level of urinary calcium. Age, sex, antihypertensive medication, drinking habit, sodium intake or order of treatment did not significantly influence the effects of calcium supplementation. CONCLUSIONS: An increase in calcium intake tends to lower office, home and ambulatory blood pressure in hypertensive patients. However, the antihypertensive effect is too small to support the general application of a high calcium intake in the treatment of hypertension. PMID- 9856372 TI - The Dundee Step Test: a novel exercise test suitable for the outpatient management of hypertension. PMID- 9856373 TI - Overestimation of the prevalence of hypertension in the population. PMID- 9856374 TI - Lack of relationship between left ventricular mass and serum cholesterol in hypertensives. PMID- 9856375 TI - Different effects of nifedipine and amlodipine on circulating catecholamine levels in essential hypertensive patients. AB - OBJECTIVE: To compare the acute and chronic effects of nifedipine retard (NPA), nifedipine gastrointestinal therapeutic system (NGITS) and amlodipine at trough and peak plasma concentrations of drug on blood pressure and heart rate, and on plasma norepinephrine and epinephrine levels in patients with mild-to-moderate hypertension (diastolic blood pressure 95-115 mmHg). DESIGN AND METHODS: After 3 4 weeks' placebo treatment, patients of both sexes were randomly allocated to be administered 10 or 20 mg NPA twice a day, 30 or 60 mg NGITS once a day, and 5 or 10 mg amlodipine once a day for 6 weeks. Initially, for the first 2 weeks, the lowest dose of each drug was used, but higher doses were administered after 2 weeks if sitting diastolic blood pressure was > 90 mmHg. Patients were evaluated after administration of the first dose and after 6 weeks' therapy in a hospital setting. Blood samples were taken for high-performance liquid chromatography measurement of catecholamine and drug levels at various intervals for a period covering trough to peak drug level ranges. RESULTS: Administration of all three drugs reduced clinic blood pressure to the same level after 6 weeks' therapy, but heart rate was increased slightly only with amlodipine (P < 0.05). Administration of NPA reduced blood pressure more abruptly whereas administrations of NGITS and amlodipine induced smoother falls after acute and chronic treatments: a significant increase in heart rate was observed with amlodipine after chronic treatment. Both acute and chronic treatments with NPA (n = 19) increased norepinephrine levels (P < 0.01) transiently (2-4 h). In contrast, administration of NGITS (n = 22) did not increase norepinephrine levels and even induced a slight but significant decrease in norepinephrine levels 5-6 h after chronic treatments. Although administration of amlodipine (n = 22) did not increase norepinephrine levels transiently either after acute or after chronic administration, it did induce a sustained rise in basal norepinephrine levels by more than 50% after chronic therapy (P < 0.01). Plasma epinephrine levels were not increased by any of the treatments and even a slight decrease was observed 4 h after administration of a dose following chronic treatments with NGITS and amlodipine (P < 0.05). CONCLUSIONS: The transient increase in norepinephrine levels observed with NPA and the sustained increases in norepinephrine levels observed after chronic treatment with amlodipine suggest that sympathetic activation occurs with those two drugs. The lack of increase in norepinephrine levels after administration of NGITS suggests that this formulation does not activate the sympathetic system. The lowering of epinephrine levels after administrations of NGITS and amlodipine suggests that inhibition of release of epinephrine by the adrenal medulla occurs with longer-acting dihydropyridine formulations. PMID- 9856376 TI - Introduction: the role of calcium antagonists. PMID- 9856377 TI - Cardiovascular risks of hypertension: lessons from observational studies. AB - Hypertension is an acknowledged major risk factor for cardiovascular disease and death in both men and women. Despite a historical focus by clinicians on the importance of diastolic blood pressure (DBP) risks, epidemiologic data from numerous large-scale studies have clearly demonstrated that both systolic blood pressure (SBP) and DBP are important determinants of cardiovascular risk. Recent analyses have described notable risks associated with isolated and borderline elevations of SBP, which predominate in the elderly, emphasizing the independent contribution of elevated SBP in determining overall risk. Overviews of large scale treatment trials show that antihypertensive drug treatment confers a favorable net clinical benefit in patients with diastolic and isolated systolic hypertension, and the magnitude of risk reduction is comparable to that expected from the observational data. However, at any level of SBP or DBP, the absolute magnitude of risk varies widely depending on the burden of coexisting risk factors present. Therefore, it is essential that decisions regarding the urgency, risks and benefits of antihypertensive drug treatments be informed by accurate determinations of overall cardiovascular risk. PMID- 9856378 TI - Calcium antagonists and sympathetic activity in congestive heart failure. AB - OBJECTIVE: To review the sympathetic abnormalities occurring in heart failure, their pathophysiological importance and clinical relevance, and the effects of drug treatment, with particular reference to calcium antagonists. SYMPATHETIC ACTIVATION IN HEART FAILURE: Indirect and direct approaches to study sympathetic function in humans have documented conclusively that sympathetic activation represents a hallmark of cardiac failure syndrome. Evidence indicates that sympathetic overactivity is associated with, and probably caused by, a baroreflex impairment and that it has adverse effects on patients' prognosis and survival. GOALS OF DRUG TREATMENT IN CONGESTIVE HEART FAILURE: In the past, drug treatment in heart failure was aimed at improving patients' survival by ameliorating cardiac hemodynamics. It is now established that a major goal of therapeutic intervention is also to reduce sympathetic activation characterizing heart failure. CALCIUM ANTAGONISTS IN HEART FAILURE: Studies with short-acting calcium antagonists show that they enhance sympathetic activation and that this has an adverse effect on patients' survival. In contrast, third generation calcium antagonists such as amlodipine, which have a slow onset and long duration of action, do not adversely affect sympathetic function and reflex cardiovascular control. Indeed, evidence suggests calcium antagonists with this profile may exert favorable clinical effects. PMID- 9856379 TI - Two-year follow-up study to evaluate the reduction of left ventricular mass and diastolic function in mild to moderate diastolic hypertensive patients. AB - OBJECTIVE: To compare the effects of two antihypertensive agents, amlodipine and lisinopril, on left ventricular mass and diastolic filling in patients from primary care centers with mild to moderate diastolic hypertension. STUDY DESIGN: A second-year, open follow-up of a prospective, double-blind, randomized, parallel group, comparative study. METHODS: Male and female patients between 25 and 75 years-of-age with elevated diastolic blood pressure (four measurements > or = 95 mmHg from multiple measurements taken on three occasions and average diastolic blood pressure < 115 mmHg) were recruited from a population survey. After 4 weeks' placebo run-in, 71 patients were included of whom 60 finished the first study year and 51 finished the second study year. Patients were randomly assigned to receive doses of 5-10 mg amlodipine or 10-20 mg lisinopril, which were titrated on the basis of the effects on blood pressure. Primary endpoints were left ventricular mass index (LVMI) and early to atrial peak filling velocity. Office and ambulatory blood pressure were considered secondary endpoints. RESULTS: The decrease in blood pressure was equal for both treatment regimens in the first year. A statistically significant (P< 0.001) decrease in LVMI in both treatment groups was observed in the first year [-11.0 g/m2 (95% Cl 6.0 to -16.1) in the amlodipine group and -12.6 g/m2 (95% Cl -8.2 to -17.0) in the lisinopril group]. Early to atrial peak filling velocity did not change significantly within the treatment groups in the first year [+0.07 (95% CI -0.01 to +0.15) in the amlodipine group and +0.01 (95%9 Cl -0.06 to +0.08) in the lisinopril group. Blood pressure, LVMI and early to atrial peak filling velocity did not change in the second year of treatment. No significant differences in primary and secondary endpoints between treatment groups were found in the first or second year. CONCLUSION: The effects of amlodipine and lisinopril on left ventricular mass and early to atrial filling peak velocity after 2 years of treatment were similar and these effects were already observed after 1 year of treatment. Additional studies of longer duration (> or = 4 years) and with a larger sample size are recommended. PMID- 9856380 TI - The Dutch Renal Artery Stenosis Intervention Cooperative (DRASTIC) Study: rationale, design and inclusion data. AB - RATIONALE: Renal artery stenosis may lead to renovascular hypertension, risking multiple organ damage including damage to the contralateral kidney. Progression of stenosis may impair the function of the affected kidney. It is important to identify individuals with this disease among hypertensive patients. The first aim of the Dutch Renal Artery Stenosis Intervention Cooperative (DRASTIC) study is to assess the prevalence of renal artery stenosis in patients with well-defined forms of drug-resistant hypertension, and to determine the predictive value of clinical characteristics and diagnostic tests in these pre-selected patients. With regard to treatment, the effect of renal angioplasty on hypertension is disappointing in atherosclerotic stenosis and technical failure frequently occurs. Therefore, the second aim is to compare the effects of balloon angioplasty and antihypertensive medication on blood pressure in patients with atherosclerotic renal artery stenosis. DESIGN HYPERTENSIVE: patients receiving standard antihypertensive medication in whom diastolic blood pressure remained > or =95 mmHg during three consecutive visits to the outpatient clinic underwent full diagnostic work-up, including renal arteriography. The prevalence of renal artery stenosis in this well-defined patient group was then established, and the predictive value of the various diagnostic tests was assessed. Patients with an atherosclerotic renal artery stenosis of > or =50% were then randomly assigned to balloon angioplasty or to treatment with antihypertensive drugs. After 1 year of intensive follow-up of blood pressure and renal function, re-arteriography was performed. CONCLUSION: In total, 1205 patients have been included in the study, about 500 have received diagnostic work-up, and it is expected that 100 patients will be randomly assigned for renal angioplasty or medical treatment. PMID- 9856381 TI - Regression of left ventricular hypertrophy in hypertension: changing patterns with successive meta-analyses. AB - Left ventricular hypertrophy (LVH) may be an important target for therapy in hypertension, although definitive data in humans are not available. An important question is whether antihypertensive drugs vary in their ability to cause the regression of LVH. Without adequately powered controlled comparisons, meta analysis provides the best method of assessing the overall results of the numerous smaller published studies. A succession of meta-analyses have indicated a strong relationship between changes in blood pressure and LVH regression. We have reviewed studies conducted for 5 years since the first major meta-analysis. Methodology was found to have improved, and the overall ranking of drug classes according to their effects on LVH regression were: calcium antagonists, angiotensin converting enzyme inhibitors, diuretics, alpha-blockers, beta blockers, and lifestyle changes. Changes from earlier reports may reflect the use of new preparations within some classes of drugs and large studies are required to define whether this will be reflected in clinical outcomes. PMID- 9856382 TI - Clinical benefits of a consistent reduction in blood pressure. AB - This paper briefly reviews the epidemiological evidence that hypertension is a major cardiovascular risk factor. It also summarizes the data from controlled intervention trials that show antihypertensive treatment to be accompanied by a reduction in cardiovascular morbidity and mortality. The inability of antihypertensive treatment to offer full protection to the hypertensive individual is then discussed, together with the therapeutic strategies to increase the benefits, particularly with respect to limiting end-organ damage and reduction of cardiovascular events. In this context, emphasis is given to the potential additional benefit conferred by control of 24-h blood pressure and to the compliance advantage of using drugs with a long duration of action. The longevity of the blood pressure lowering effect can compensate for delayed or missed drug consumption, a frequent phenomenon in the chronically treated hypertensive patient. PMID- 9856383 TI - The effect of the Hurnik-Morris (HM) system on sow reproduction, attrition, and longevity. AB - Sows were randomly selected and assigned to either the Hurnik-Morris system (HM; n = 59) or a conventional gestation crate system (GC; n = 90) before breeding. The HM system provides housing for sows in small groups. All sows were bred to Duroc x Hampshire commercial boars. A common farrowing barn with 18 farrowing crates was used for the HM and GC sows. The HM sows had less (P<.001) backfat at farrowing. Average parity per sow (P<.046), lifetime number of pigs born per sow (P<.02), lifetime number of pigs born alive per sow (P<.02), and lifetime number weaned per sow (P<.045) were higher for sows housed in the HM system. The total number of sows removed from the GC system was higher (P<.025) than the number leaving the HM system. The higher parity level and lifetime production exhibited by the sows in the HM system indicates that this system supports greater longevity and may reflect the superior effect that this housing system has on animal well-being. PMID- 9856384 TI - Toward a pluralistic animal science: postliberal feminist perspectives. AB - Women have not had the authority to define the conditions of animal agriculture or to develop strategies for addressing its problems because the field of animal science has been and remains dominated by men. Therefore, criteria for assessing the professional performance of animal scientists reflect the views of men and the experiences of masculinity. Recent attempts to improve the retention rates of women in the animal science profession have focused on socializing female proteges to the norms and expectations of the discipline. This approach is based on traditional liberal beliefs that all humans are essentially alike and that embodiment and(or) social location are irrelevant in understanding the world. Such socialization assumes that problems of gender inequity can be solved by providing opportunities for women to succeed in the existing system, with its existing norms. This approach ignores the possibility that the structure of women's lives and the normative commitments arising from women's experiences may be quite different from those of men. Professional reward structures that favor the life experiences and perspectives of men and ignore those of women coerce women into adopting values that may be incompatible with their identities. This not only devalues their perspectives and contributions, it forces them to commit self-betrayal. This is both harmful and antithetical to the idea of an empirically relevant science that blends multiple perspectives and understands diverse values. Substantive diversity in the animal science community requires standards that reflect and accommodate the pluralism in which we live. PMID- 9856385 TI - Effects of feeder type, space allowance, and mixing on the growth performance and feed intake pattern of growing pigs. AB - The effect of crowding and mixing on growth performance and feed intake pattern were investigated in growing pigs in a 4-wk study. Feeding pattern was monitored using automated feed intake recording equipment (F.I.R.E.). A total of 256 Yorkshire x Hampshire and purebred Duroc pigs (initial weight 35.8+/-.86 kg) were allocated to one of the eight treatment combinations in a 2x2x2 factorial arrangement (feeder type [conventional feeder vs F.I.R.E. feeder], space allowance [.56 vs .25 m2/pig], and mixing strategy [mixed vs unmixed; mixing at start of wk 1 and 3]). Pigs were housed in groups of eight, balanced for genotype and sex (barrows and gilts), and had free access to a corn-soybean meal diet (17% crude protein, 3,296 kcal ME/kg). There was no difference in growth performance between feeder types. Crowding and mixing had no effect on daily feed intake but they depressed growth rate by 15.7 and 7.1%, respectively, and the effects of the two stressors were additive. Gain:feed ratio was reduced by crowding (10.0%) but not by mixing. Crowded pigs made fewer (11.2 vs 15.7; SEM = .51), and longer (12.5 vs 8.9 min; SEM .41) feeder visits and had higher feed intake per visit (196.2 vs 145.5 g; SEM = 5.94) than uncrowded animals. Mixing produced changes in feeding pattern in the 1st wk after mixing but not over the 4-wk period. This study showed that crowding and mixing depressed growth rates in an additive manner and altered feeding behavior. PMID- 9856386 TI - Activation state of muscle satellite cells isolated from steers implanted with a combined trenbolone acetate and estradiol implant. AB - Muscle satellite cells were isolated from seven yearling steers implanted for 31 d with a combined implant that contained 120 mg of trenbolone acetate (TBA) and 24 mg of estradiol (E2) and from seven nonimplanted, control steers. Implanted steers had a 28% greater ADG and a 23% greater feed efficiency than did nonimplanted steers. Implanted steers had increased (P<.001) circulating IGF-I concentrations on d 6, 14, and 31 after implantation, and circulating IGF-I concentrations in control steers remained constant or decreased (P<.05) at these times. Maximum fusion percentage was greater (P< .005) in satellite cell cultures isolated from implanted steers (ISC cultures) than in satellite cell cultures isolated from control steers (NSC cultures) (72.8% vs 54.8%, respectively). Satellite cell cultures isolated from implanted steers (ISC cultures) also contained a greater (P<.001) number of myotube nuclei than did NSC cultures (7,998 nuclei/cm2 vs 5,150 nuclei/cm2, respectively). After 72 h in culture, the number of cells (corrected for plating density) was 43% greater (P<.05) in ISC cultures than in NSC cultures. [3H]Thymidine incorporation rates per 10(5) cells at 24 and 34 h after plating were greater (P<.05) in ISC cultures than in NSC cultures; however, incorporation rates did not differ at 72 h. These data indicate that TBA + E2 implantation may result in an in vivo activation of muscle satellite cell proliferation that can be detected in cell culture. This activation may play an important role in TBA + E2-enhanced muscle growth. PMID- 9856387 TI - Effects of cereal type and feed particle size on morphological characteristics, epithelial cell proliferation, and lectin binding patterns in the large intestine of pigs. AB - The present study was undertaken to investigate the effects of cereal type and feed particle size on the morphological characteristics and epithelial cell proliferation of the large intestinal tissue in pigs. Forty pigs, weighing approximately 30 kg, were fed diets containing either coarsely or finely milled barley or wheat for a period of 4 wk. Tissue samples were taken from the cecum and from the proximal, medial, and distal colon at slaughter. The pigs fed the coarse diets had significantly larger crypts, in terms of height as well as volume, than did pigs fed the fine diets. The cereal type had no effect on the mucosal architecture. The epithelial cell proliferation, in terms of counted native mitoses in the crypts, was significantly higher in pigs fed the coarse barley diet than in pigs fed the coarse wheat diet or the fine diets. The volume of the mucin granules in the crypts constituted from 32 to 52% of the crypt volume and was greatest in the pigs fed the coarse diets. This effect of feed particle size was observed for neutral as well as for acidic mucins and sulfomucins. Lectin binding patterns indicated that more of the terminal sugars on glycoconjugates of the apical membrane on the mucosal surface were the sialic acid alpha-2,3 neuraminic acid, but less were mannose in the pigs fed the coarse barley diet. Distinct regional differences were observed among the intestinal sites. These included a decline in the epithelial cell proliferation and an increase in the volume of mucin in the crypts along the intestinal tract. Furthermore, the sialic acid alpha-2,3 neuraminic acid was more abundant in the medial colon than in the cecum; the contrary was seen for mannose and galactose. This study shows that the feed particle size of barley and wheat diets, more than the cereal type itself, affects the mucosal architecture, epithelial cell proliferation, and production and composition of the mucins in the large intestine of pigs. The study suggests that pigs fed a coarse diet are better protected against intestinal infections than pigs fed a fine diet. PMID- 9856388 TI - Effects of repeated urea dilution measurement on feedlot performance and consistency of estimated body composition in steers of different breed types. AB - Steers (20 Bos indicus cross [BIX] and 20 Bos taurus cross [BTX]) were randomly assigned to a 2x2 factorial experiment within two weight blocks per treatment 1) to study the effects of repeated urea dilution (UD) measurement on feedlot performance and 2) to determine the consistency of estimated body composition in steers of different breed types. Weights were taken on d 0, 42, 84, 126, and 140. Urea dilution was determined on half of the pens in the experiment, and ultrasonic measurement of backfat (BF) was performed on all cattle on d 0, 42, 84, and 126. Pen means of all performance variables were used in the analysis of variance. Carcass data were analyzed on an individual basis. Within periods, ADG was inconsistent between controls and steers on which UD was determined (1.95 vs 2.03, 1.61 vs 1.28, 1.51 vs 1.71, and 1.77 vs 1.47 kg, P = .23, .02, .09, and .11, respectively, for Periods 1, 2, 3, and 4, SEM = .07). Overall, UD had no effect (control vs UD, respectively) on ADG (1.70 vs 1.68 kg, P = .77, SEM = .07), DMI (8.26 vs 8.03 kg, P = .69, SEM = .36), gain efficiency (207 vs 209 g BW gain/kg DMI, P = .78, SEM = 2.34), hot carcass weight (HCWT; 360 vs 358 kg, P = .90, SEM = 2.52), or percentage of estimated carcass fat, (ECF; 38.8 vs 37.0%, P = .61, SEM = 1.05). Breed types (BIX vs BTX, respectively) had similar ADG (1.74 vs 1.64 kg, P = .27, SEM = .14), DMI (7.96 vs 8.30 kg, P = .50, SEM = .36), backfat thickness (16.4 vs 15.0 mm, P = .30, SEM = .45), and ECF (38.9% vs 36.6%, P = .48, SEM = 2.01). Urea dilution estimated empty body fat values increased with days on feed (14.4+/-1.36; 22.7+/-1.47; 26.0+/-1.36; 30.4+/-1.47%, respectively, for d 0, 42, 84, and 126). Using yield grade factors to calculate ECF consistently produced a value that was higher than empty body fat determined by UD (UDEBF) 14 d prior to slaughter (36.9+/-1.73 vs 30.4%+/-0.17). Significant correlation coefficients were found for the pooled data between UDEBF vs BF, r = .84; UDEBF vs live weight, r = .99; UDEBF vs ECF, r = .82; and UDEBF vs percentage of carcass protein, r = -.99. This study demonstrated that there are no detrimental effects of the urea dilution procedure on performance characteristics of feedlot cattle. Beef cattle of different breed types may be accurately evaluated with urea dilution. PMID- 9856389 TI - Cooking and palatability traits of beef longissimus steaks cooked with a belt grill or an open hearth electric broiler. AB - The objective of this experiment was to compare the effects of belt grill and Open Hearth electric broiler cookery on palatability and cooking traits of longissimus steaks. The longissimus thoracis from carcasses of grain-fed steers or heifers was used. Duplicate measurements were made for Warner-Bratzler shear force at 3 and at 14 d after slaughter (n = 180) and trained sensory evaluation at 14 d after slaughter (n = 91) using both cooking methods. Belt grill-cooked samples had lower (P<.01) percentage of cooking losses (21.5 vs 25.8%) and higher (P<.01) shear force values (4.6 vs 4.3 kg) than electric broiler-cooked samples. Repeatability of duplicate measurements was higher for cooking losses (.58 vs .23) and shear force values (.85 vs .64) for belt grill than for electric broiler cooked samples. Belt grilled steaks had lower (P<.01) cooking losses (20.2 vs 29.8%); higher (P<.01) tenderness (7.0 vs 6.7) and juiciness (6.0 vs 5.1); and lower (P<.02) connective tissue amount (7.7 vs 7.8), beef flavor intensity (5.0 vs 5.1), and off-flavor (3.2 vs 3.3) ratings than steaks cooked with the electric broiler. Belt grill cooking increased the repeatability of duplicate sensory measurements for tenderness (.87 vs .71), connective tissue amount (.66 vs .30), and juiciness (.51 vs .08) ratings, and cooking losses (.63 vs .18) compared with cooking with the electric broiler. Belt grill cooking increased the precision for measurements of cooking, Warner-Bratzler shear force, and palatability traits of beef longissimus thoracis. PMID- 9856390 TI - Effects of postmortem storage on the ultrastructure of the endomysium and myofibrils in normal and callipyge longissimus. AB - These experiments were conducted to examine ultrastructural changes in longissimus from normal and callipyge lamb during 14 d of postmortem storage at 4 degrees C. Six crossbred ewe lambs (1/2 Dorset x 1/2 Romanov) were grain-fed and slaughtered at approximately 250 d of age. Leg conformation score was the basis for classifying carcasses into normal and callipyge. The normal and callipyge longissimus had mean Warner-Bratzler shear force of 2.8 (2.7, 2.4, and 3.4) and 9.0 (12.2, 6.9, and 7.9) kg, respectively, after 14 d of postmortem storage. The results of transmission electron microscopy demonstrated ultrastructural changes, including sarcolemma detachment, loss of myofibril lateral attachments, and I band breaks in normal longissimus. Detachment of sarcolemma from myofibrils occurred in both phenotypes, but it was delayed by several days in callipyge longissimus. Thus, the sarcolemma detachment seems not to contribute significantly to postmortem tenderization. The endomysium of both phenotypes did not change with postmortem storage. In normal longissimus, the percentage of fractured I-bands increased from 0% at d 1 to 11% at d 3 (P<.05) and did not change between 3 and 14 d (15%) postmortem (P>.05). However, postmortem storage did not affect (0 to 3%) the frequency of the I-band breaks in the callipyge longissimus (P>.05). Therefore, the break in the I-band region in postmortem muscle is a change that is associated with postmortem tenderization. We conclude that the major factor responsible for the toughness of meat from callipyge longissimus is the postmortem stability of myofibrils. PMID- 9856392 TI - Limiting order of amino acids in a low-protein corn-soybean meal-whey-based diet for nursery pigs. AB - Three trials were carried out with pigs between 5 and 8 wk of age to determine the limiting order of amino acids in a 13.5% CP corn-soybean meal-based diet containing 8% dried whey. The positive-control diet was a 19.2% CP corn-soybean meal-based diet (1.15% lysine), also with 8% dried whey. Amino acid additions to the low-protein, negative-control diet were based on levels needed to accomplish 110% of ideal ratios (to lysine, set at 1.15%). In Exp. 1, the addition of an amino acid mixture containing Lys, Trp, Thr, Met, Ile, and Val to the low-protein diet increased (P<.05) gain and gain: feed ratio, and these response traits were not different from those of pigs fed the 19.2% CP positive-control diet. Single deletion of Lys from the supplemental amino acid mixture depressed performance to a greater (P<.05) extent than single deletion of any of the other amino acids. Single deletions of Trp, Thr, Met, or Val decreased (P<.05) performance in a similar but lesser magnitude than the decrease caused by Lys deletion, whereas Ile deletion was without effect. Experiments 2 and 3 were designed to evaluate the limiting order of AA beyond Lys in the low-protein diet. Neither His nor Glu were found to be deficient, and, as in Exp. 1, deletion of Trp, Thr, Met, or Val from the supplemental amino acid mixture resulted in performance depressions (P<.05) that were similar. The results suggest that Lys is first-limiting and Trp, Thr, Met, and Val are equally second-limiting in a reduced protein (13.5% CP) corn-soybean meal-based diet with 8% whey for 10-kg pigs. PMID- 9856391 TI - Effect of dietary fiber or fat in low-crude protein, crystalline amino acid supplemented diets for finishing pigs. AB - Three experiments were conducted to determine the effect of reducing NE, by adding dietary fiber in Exp. 1 and 2 and decreasing dietary fat in Exp. 3, of low CP, crystalline amino acid (CAA)-supplemented diets for finishing pigs on growth performance and carcass characteristics. In Exp. 1 and 2, 64 barrows (Exp. 1) or gilts (Exp. 2) were allotted to four treatments with four replicates of four pigs each. Average initial and final BW were 74 and 117 kg in Exp. 1 and 74 and 102 kg in Exp. 2. The following diets were fed in Exp. 1: 1) corn-soybean meal (C-SBM); 2) low-CP (-3.5%), supplemented with CAA; 3) CAA + rice hulls (CAA+RH; NE equal to Diet 1); and 4) CAA+RH+OIL (NE equal to Diet 2). Experiment 2 was similar to Exp. 1, except RH were replaced with wheat middlings (WM), oil was replaced with dry fat, and the CP was decreased by 3.1% in the low-CP diets. In both experiments, serum urea-N (SUN, corrected for initial SUN by covariance analysis) was higher (P<.10) for pigs fed C-SBM than for pigs fed any other diet. In Exp. 1, barrows fed CAA+RH had lower hot carcass weight, percentage muscle, fat-free lean (FFLEAN), lean gain per day, retained energy (RE) in FFLEAN, and lean:fat ratio than barrows fed C-SBM, along with less FFLEAN than barrows fed CAA+RH+OIL. Barrows fed CAA+RH had smaller longissimus muscle areas than barrows fed any other diet, and barrows fed C-SBM had higher dressing percentage and lower percentage total fat than barrows fed any other diet. Barrows fed C-SBM had higher lean:fat ratio and lower total fat than barrows fed CAA. In Exp. 2, gilts fed CAA+WM+FAT had heavier heart weights than gilts fed C-SBM or CAA (P<.10). In Exp. 3, 702 gilts were allotted to six treatments with nine replicates of 13 gilts each. Average initial and final BW were 70 and 110 kg. Gilts were fed two levels of CP (15.5 or 11.7% plus CAA added to meet an ideal amino acid ratio) and three levels of NE (2,650, 2,617, or 2,584 kcal/kg), resulting in a 2x3 factorial arrangement of treatments. Gilts fed 15.5% CP had higher gain:feed ratio than gilts fed 11.7% CP (P<.01). Longissimus depth was greater for gilts fed 15.5% CP than for gilts fed 11.7% CP and was decreased as NE decreased only in gilts fed 11.7% CP (CP effect, P<.09; NE linear effect, P<.04; CP x NE effect, P<.01). Gilts fed the diet with 2,617 kcal NE had lighter carcasses and less total fat, fat gain per day, RE, and RE as fat regardless of protein level than gilts fed 2,650 or 2,584 kcal NE/kg (NE quadratic, P<.09). Loin color score increased as NE decreased (linear, P<.06), but longissimus fat depth was increased by the lowest level of NE (NE quadratic effect, P<.09). Overall, the reduction of NE in low-CP, CAA-supplemented diets did not affect growth performance and was not an effective means of reducing fat in finishing pigs. PMID- 9856393 TI - Evaluation of enzymatically modified potato starches in diets for weanling pigs. AB - We conducted three growth trials to evaluate replacing carbohydrate sources with enzymatically modified potato starches in diets for weanling pigs. In Exp. 1, 180 pigs (initially 5.3 kg and 21+/-2 d of age) were used to compare the effects of corn (36.5%), edible-grade oat flour (36.5%), two enzymatically modified potato starches (12%), and added lactose (12%) on pig performance. Potato Starch 1 had a dextrose equivalent (DE) of 6 and Potato Starch 2 had a DE of 20; both were spray dried maltodextrans. Pigs that were fed Potato Starch 2 had greater (P<.05) ADG and ADFI than pigs fed diets that contained corn or oat flour from d 0 to 14 after weaning, and pigs fed either Potato Starch 1 or added lactose had intermediate ADG and ADFI. However, for the overall trial (d 0 to 35), no differences (P>.10) in growth performance were observed. In Exp. 2, 198 pigs (initially 4.3 kg and 19+/-2 d of age) were used to determine whether modified Potato Starch 2 could replace a portion of the corn or lactose in the diet. The control diet contained 10% dried whey, and additional treatments were formulated by adding 7 or 14% modified Potato Starch 2 or lactose in place of corn. A positive control diet was formulated containing 29% dried whey. From d 0 to 14 after weaning, increasing dietary lactose improved (linear, P<.04) ADG and ADFI. Increasing the potato starch had no effect on ADG but increased ADFI (linear, P<.02). In Exp. 3, 180 pigs (initially 3.9 kg and 14+/-3 d of age) were used to evaluate Potato Starch 2 or 3 (DE = 30, a spray-dried glucose syrup) as replacements for either corn or lactose in the diet. Pigs were fed a control diet containing 15% dried whey and 12% added lactose. Twelve percent modified Potato Starch 2 or 3 replaced either corn or lactose in the diet on a wt/wt basis. From d 0 to 14 and d 0 to 21, pigs fed either modified potato starch substituted for corn had greater (P<.07) ADG than those fed the control diet. Pigs fed diets with either modified starch substituted for lactose had similar ADG as those fed the control diet. These results suggest that when substituted for corn, Potato Starch 2 can improve growth performance of weanling pigs. PMID- 9856394 TI - Nutritional evaluation of two agroindustrial by-products for ducks and pigs. AB - Forty-eight male White Pekin ducks and 24 crossbred barrows (Landrace or Yorkshire sows; PIC Line 355 boars) were used in three experiments to evaluate the nutritional value of two by-products generated from agricultural industries. In each experiment, test ingredients consisted of a dried meat solubles product (DMS) from swine processing facilities and a yeast fermentation product (YFP) from a commercial source. Soybean meal (SBM) with a CP content of 48% served as the control ingredient in the three experiments. Different batches of each by product were evaluated in the three experiments utilizing an apparent ME (AME) assay for poultry (Exp. 1 and 2) and a digestibility assay for barrows (Exp. 3). The nitrogen-corrected AME (AMEn) for SBM, DMS, and YFP in Exp. 1 were 2.909, 2.801, and 3.292 kcal, respectively. The AMEn for SBM, DMS and YFP in Exp 2 were 2.809, 3.207, and 3.565 kcal, respectively. In Exp. 3, diets were formulated such that the test ingredients provided the sole source of amino acids in each experimental diet. The AME of SBM, DMS, and YFP in Exp. 3 were 3.844, 2.208, and 3.552 kcal, respectively. Based on the results of the ME assay for poultry, the DMS product compares well to SBM and seems to be suitable for feeding to ducks. The YFP does not seem to be suitable for feeding nonruminant species based on poor ability to support nitrogen retention in both species (-36% and -20% in Exp. 1 and 2, and 20% in Exp. 3). PMID- 9856395 TI - Endophytic fungal toxin effect on adrenergic receptors in lateral saphenous veins (cranial branch) of cattle grazing tall fescue. AB - The objective of this research was to characterize the adrenergic receptor response in veins of cattle that grazed tall fescue (Festuca arundinacea Schreb.), with (E+) and without (E-) fungal endophyte (Neotyphodium coenophialum) infection. Measurement of contractile response of lateral saphenous veins to selective adrenergic receptor agonists (alphal, phenylephrine; alpha2, BHT-920) revealed enhanced reactivity (greater contractile response) only for the alpha2 adrenergic receptor in E+ pastured cattle. This response was consistent over a 3 yr grazing period. Significant contractility differences in response to BHT-920 occurred at concentrations of 3x10(-6) M and greater. Early research has reasoned that the enhanced reactivity is due to endophyte toxin effect on intracellular signaling systems. The enhanced contractile effect would alter blood flow dynamics and contribute to clinical signs and pathologic change in animals. The alpha2-adrenergic receptor is also involved in the control of many metabolic reactions. Thus, for drug therapy to be successful in reversing the adverse effects of E+ tall fescue exposure in cattle, effects on the alpha2-adrenergic tissue receptors must be neutralized. PMID- 9856396 TI - In vitro effect of swainsonine on bovine and ovine lymphoblastogenesis. AB - Peripheral blood lymphocytes (PBL) were obtained from heifers and wethers to assess the in vitro effects of swainsonine. Stimulated and unstimulated PBL (2x10(5)/well; 96 well plates) were incubated in the presence of swainsonine (2, .2, .02, .002, and 0 microg/mL; n = 32) for 72 h. Mitogens used to stimulate PBL were concanavalin A (ConA; 5 microg/mL), phytohemagglutinin-P (PHA-P; 5 microg/mL), phytohemagglutinin-M (PHA-M; 5 microg/mL), pokeweed mitogen (PWM; 5 microg/mL), and lipopolysaccharide (LPS; 10 microg/mL). Controls received no swainsonine or mitogen. Effects of swainsonine are expressed as a percentage of stimulation relative to control (stimulation index; SI). Unstimulated bovine PBL treated with 2 microg/mL of swainsonine exhibited a depressed (P<.001; SI of 87 and 100 for 2 microg/mL and control, respectively; SE = 2.7) SI. At concentrations of .2 and 2 microg/mL, swainsonine inhibited (P<.001; SI of 351, 310, and 464 for .2, 2, and 0 microg/mL, respectively; SE = 27.1) bovine PBL proliferative response to PHA-P. Swainsonine had a mitogenic effect on unstimulated ovine PBL at .2 and 2 microg/mL (P<.05; SI of 118, 113, and 100 for .2 and 2 microg/mL and control, respectively; SE = 4.4). Swainsonine inhibited ovine proliferative responses to PHA-P at .2 and 2 microg/mL (P<.005; SI of 190, 178, and 228 for .2, 2, and 0 microg/mL, respectively; SE = 9.4) and to PHA-M at .002 and 2 microg/mL (P<.03; SI of 165, 167, and 192 for .002, 2, and 0 microg/mL, respectively; SE = 7.9). The opposing responses of unstimulated ovine (mitogenic) and bovine (antiproliferative) PBL to swainsonine indicates a differential species response. Swainsonine suppression of PHA-P-induced proliferation would indicate a negative effect on ovine and bovine T-cell function. PMID- 9856397 TI - Effects of fumonisin B1-contaminated feeds on weanling angora goats. AB - Two diets containing no (<1.0 mg/ kg) or 95 mg of fumonisin B1 (FB1)/kg were fed to eight weanling Angora goats for 112 d. Dry matter intake, apparent nutrient digestibilities, serum chemistry profiles, sphingolipid concentrations, and persistency of FB1 in tissues were evaluated. No differences (P>.10) were found between control and treated goats in terms of DMI, apparent nutrient digestibilities, or ADG. Elevated concentrations (P<.10) of blood-borne enzymes such as aspartate aminotransferase, lactate dehydrogenase, and gamma glutamyl transpeptidase and increased concentrations of cholesterol and triglycerides indicated mild liver damage and kidney dysfunction in treated goats. Linear relationships (P<.10) were observed between these serum constituents and duration of FB1 exposure. The sphingolipid analysis of liver, kidney, and heart tissues showed elevated free sphinganine:free sphingosine ratios in the treated group. The elevated sphingolipid ratios were mainly due to increased concentrations of free sphinganine in tissues. However, without serum profile and sphingolipid analyses, fumonisin toxicosis would not have been recognized because treated animals showed no clinical signs of toxicosis throughout the trial. No measurable FB1 was present in liver, kidney, and heart tissues (detection limit of 1 ppm). However, further research is needed to analyze tissues for FB1 or its metabolites with a lower detection limit. In conclusion, goats can be fed for up to 112 d with diets containing 95 mg FB1/kg of diet without any overt signs of toxicosis and also without any effect on live weight gain. PMID- 9856398 TI - Effects on performance, tissue integrity, and metabolism of vitamin E supplementation for beef heifers fed a diet that contains gossypol. AB - We conducted an experiment for 112 d with yearling beef heifers to evaluate the effects of cottonseed meal (CSM) fed with various concentrations of vitamin E on hematological and tissue components. Heifers were assigned randomly to four treatments, with eight heifers per treatment. The treatments consisted of the following dietary supplements: 1) CON, based on soybean meal with 30 IU vitamin E/kg; 2) GOS, based on CSM with 30 IU vitamin E/kg; 3) G+2E, based on CSM with 2,000 IU vitamin E x animal(-1) x d(-1); and 4) G+4E, based on CSM with 4,000 IU vitamin E x animal(-1) x d(-1). Supplements based on CSM provided 4.5 g of free and 50.5 g of total gossypol x animal(-1) x d(-1). The total gossypol present in the supplements was 29.1% of the negative isomer (-) and 70.9% of the positive isomer (+). Blood samples were collected at the start of the experiment and every 2 wk thereafter up to 16 wk. There was a time x treatment interaction (P<.01) for plasma alpha-tocopherol ( alpha-T) concentration; however, feeding gossypol did not decrease plasma alpha-T. Weight gain, retinol palmitate, retinol, beta carotene (beta-C), hemoglobin, and hematocrit were not affected by treatment. Erythrocyte osmotic fragility (EOF) increased (P<.05) in gossypol-fed animals; however, vitamin E supplementation lowered EOF (P<.05). Heifers fed the supplements GOS, G+2E, and G+4E had greater (P<.01) plasma (-)-, (+)-, and total gossypol than heifers fed CON from Collection 2 to the end of the experiment. There was a treatment effect (P<.05) on vitamin E and gossypol concentrations in different tissues, with no effect (P>.05) for trace minerals (Cu, Zn, Fe, and Se). Vitamin E concentration in tissue increased with increased dietary supplementation of vitamin E. In heart and neck muscle, (-)-gossypol was greater (P<.05) than (+)-gossypol, but the reverse was true for liver. Gossypol decreased in vitro lipid peroxidation of liver homogenate in tissues. Gossypol deposition in tissue was liver > heart > muscle. In summary, gossypol from CSM did not decrease concentrations of antioxidant vitamins, including alpha-T, vitamin A, and beta-C, or have any detrimental effect on performance of beef heifers. PMID- 9856399 TI - Evaluation of selected mathematical approaches to the kinetics of protein degradation in situ. AB - A linear model, two mathematical nonlinear models, and a curve-peeling procedure were used to estimate rate and extent of ruminal CP degradation of meat and bone meal (MBM) and soybean meal (SBM) from data obtained using the in situ Dacron polyester bag technique. Most of the values for extent of CP degradation of MBM were lowest when determined using curve peeling or the nonlinear models. In general, rates and extents of CP degradation of MBM estimated using the linear model and including ruminal incubations up to 12 h were greater than those obtained with the linear model and including ruminal incubations up to 24 h or up to 72 h. In addition, the models ranked the MBM samples differently for rate and extent of CP degradation. The results of the lack-of-fit test indicated that the linear model was inappropriate for estimating rate of degradation of MBM. However, CP degradation for SBM could be described by the linear model if long ruminal incubation times (greater than 48 h) were included in the calculations. Regression analyses were conducted to evaluate various compositional characteristics as predictors of CP degradation for MBM. Most of the correlation coefficients between CP degradation and the same independent variables were greater when the nonlinear models and curve peeling were used compared with the linear model. In general, the correlation coefficients between extent of CP degradation and the independent variables obtained with the linear model increased as the duration of ruminal incubations included in the model increased. Lysine concentrations, followed by CP solubility and ash content, were the best predictors of ruminal degradation of MBM protein. When using a specific mathematical model to predict CP degradation, analysis of residuals vs fitted and lack-of-fit tests should be performed to assess the validity of the model to describe the degradation patterns of the protein source under consideration. Also, long (at least 48 h) ruminal incubation times may be needed to correctly describe the pattern of CP degradation for MBM. PMID- 9856400 TI - Reproductive effects of feeding gossypol and vitamin E to bulls. AB - An experiment was conducted to determine the effects of long-term feeding of cottonseed meal on the reproductive traits of Holstein bulls. Holstein bulls approximately 6 mo of age were placed on the following treatments: 1) soybean meal + corn (CON); 2) cottonseed meal + corn (GOS); and 3) cottonseed meal + 4,000 IU vitamin E x bull(-1) x d(-1) (G+4E). The GOS and G+4E diets were formulated to supply 14 mg of free gossypol x kg(-1) BW x d(-1). These bulls had been in a previous experiment that evaluated the effects of feeding the same type of diets, but from 2 wk to 6 mo. of age. Percentage of motility, percentage of normal and live sperm, and daily sperm production were less (P<.05) in the GOS than in the other two treatments. Percentages of primary abnormalities and abnormal midpieces were greater (P<.05) in the GOS group than in the other two groups. At 12 and 16 mo. of age, bulls were given two assessments for sex drive traits. Bulls that received gossypol exhibited less sexual activity (P<.05) at the first test than bulls in other treatments. Vitamin E supplementation in bulls that received gossypol improved the number of mounts in the first test and the time to first service in the second test. There was a trend of gossypol to decrease and vitamin E to improve libido score. The results of the GOS first libido test may indicate lack of sexual maturity, which agrees with sperm production data. At the time of first test (12 mo. of age), none of GOS, two of CON, and six of G+4E bulls had reached puberty on the basis of experimental protocol. Long-term feeding of gossypol to Holstein bulls negatively affected some reproductive traits; however, vitamin E supplementation countered these adverse effects and even improved these traits. PMID- 9856402 TI - Fibrolytic enzyme treatment of barley grain and source of forage in high-grain diets fed to growing cattle. AB - We conducted a study to determine the effects of treating barley grain with a fibrolytic enzyme mixture on chewing activities, ruminal fermentation, and total tract digestibility in cattle. We also investigated the potential benefits of using barley straw rather than barley silage as a roughage source in high-grain diets for feedlot cattle. Steers were given ad libitum access to one of four diets that consisted of 95% barley-based concentrate and 5% forage (DM basis). The concentrate was either control or enzyme-treated, and the forage was either barley silage or barley straw. Applying the enzyme mixture onto the barley lowered the concentrations of dietary ADF and NDF. However, it is not certain when this fiber hydrolysis occurred relative to feed consumption because the fiber analyses were conducted after the study was completed. Enzyme treatment of barley increased total tract dietary ADF digestibility by 28% (P<.05). Acetate-to propionate ratio tended to decrease, which suggests that enzymes may have increased ruminal starch digestion as a result of enhanced digestion of barley hulls. Replacing silage with straw increased ADF intake (P<.05) and resulted in 1 h/d increase in rumination time (P<.05). Even though there was no effect of diet on ruminal pH, replacing silage with straw increased ruminal acetate, as a percentage of total VFA, and total tract ADF digestion (P<.01). This study demonstrates that using a fibrolytic enzyme mixture in high-grain diets that contain mainly barley grain can improve fiber digestion and grain utilization, but the mode of action is unclear. Straw can be used rather than silage to increase the effective fiber content of a high-grain feedlot diet. PMID- 9856401 TI - Effect of ruminal cellulolytic bacterial concentrations on in situ digestion of forage cellulose. AB - To evaluate the effects of ruminal cellulolytic bacterial concentrations on in vivo cellulose digestion, varying percentages of flaked soybean hulls were substituted for orchardgrass hay in high-forage diets fed to sheep. In two experiments, total and cellulolytic ruminal bacterial concentrations were not affected by diet. No differences were found for in situ digestion of forage cellulose in the first experiment; however, in Exp. 2, ruminal pH and in situ cellulose digestion were lower (P<.01) with a 40% soybean hull diet. In Exp. 3 with four sheep, two diets were compared, one containing 19.6% cellulose from alfalfa meal and the other 64.3% purified wood cellulose. Ruminal pH was lower (P<.02), 9 and 24 h after feeding, for the high-cellulose diet. Total bacterial concentrations did not change with diet; however, the concentration of cellulolytic bacteria increased (P<.05) when the higher cellulose diet was fed. In situ cellulose digestion was not different between diets. In Exp. 4, 3% sodium bicarbonate was added to the high-cellulose diet, and it was fed twice a day. No differences were observed in pH between diets (P>.42). However, the concentration of total ruminal bacteria increased (P<.06), the concentration of cellulolytic bacteria increased (P<.03), and the percentage of cellulolytic bacteria increased (P<.04) when the buffered high-cellulose diet was fed. In situ digestion of alfalfa cellulose at 30 h was not different between diets (P>.60). These data indicate that the concentration of cellulolytic bacteria is not the limiting factor in the digestion of cellulose in the rumen. PMID- 9856403 TI - Glucose transporter gene expression in lactating bovine gastrointestinal tract. AB - Absorption of glucose from the intestine and probably from various portions of the gastrointestinal tract (GIT) in lactating dairy cows can have profound implications because the availability of glucose, or glucose precursors, seems to be a limiting factor for milk synthesis. This article reports on the expression of the Na+/glucose cotransporter gene (SGLT1) in mucosal tissues from rumen, omasum, abomasum, duodenum, jejunum, ileum, and cecum of lactating cows. Sodium/glucose cotransporter mRNA transcripts were detected in polyadenylated RNA of all the GIT tissues examined and in total RNA from rumen, omasum, duodenum, jejunum, ileum, and cecum. The pattern of SGLT1 mRNA distribution in these tissues was different from that of the facilitative glucose transporter GLUT5. Sodium/glucose cotransporter protein was also detected in small intestinal membranes of lactating dairy cows. Sodium-dependent glucose uptake across small intestinal brush border membrane vesicles could be resolved into linear nonsaturable (diffusion) and saturable, carrier-mediated components. The diffusion constant of glucose from jejunal membrane vesicles was 18.3+/-2.1 pmol x mM(-1) x mg protein(-1) x s(-1). The saturable component of glucose uptake had a Vmax of 27.1+/-5.8 pmol x mg protein(-1) x s(-1) and a Km of .12+/-.05 mM. Similar transport kinetics were found in the duodenum, and they were less in the ileum. These data suggest that the GIT of lactating cows is capable of active transport of glucose from the lumen across the brush border membrane of the epithelial cells. In addition, the relatively high abundance of SGLT1 mRNA in the rumen, omasum, and cecum may imply new functional and nutritional roles for these tissues in the active transport of glucose. PMID- 9856404 TI - Influence of intravenous L-carnitine administration in sheep preceding an oral urea drench. AB - Two experiments were conducted to investigate the effect of i.v. administration of L-carnitine on selected metabolites in sheep and to determine the feasibility of using L-carnitine to ameliorate the deleterious effects of hyperammonemia in sheep. In Exp. 1, i.v. L-carnitine solutions were administered at three levels in a replicated Latin square: 0 (CONT), 6.36 (CAR 1), and 12.72 (CAR 2) mmol L carnitine/kg x (75) BW using Suffolk ewes (n = 6; average BW 75+/-3 kg). Plasma L carnitine concentration was increased (P<.05) by treatment (51.9 vs 102.3, and 96.4 micromol/L in CONT, CAR 1, and CAR 2, respectively). Plasma glucose concentration was elevated (P<.05) in CAR 2 and CAR 1. Plasma NEFA concentration was highest (P<.05) in CAR 2. Area under the response curve for glucose was greater (P<.02) in CAR 2. In Exp. 2, Suffolk ewes (n = 16; average BW 48+/-2 kg) were used in a randomized complete block design with a 2x2 factorial treatment arrangement to determine the effects of i.v. L-carnitine administration during an oral urea load test (OULT). L-Carnitine (0 and 6.36 mmol/kg x (75) BW) was administered i.v. at 30 min, and an oral urea drench (50% wt/vol; 0 and 300 mg/kg BW) was administered at 60 min. Plasma L-carnitine was increased (P<.0001) by i.v. L-carnitine. Plasma ammonia N was highest (P<.0001) in the UREA treatment compared with the CONT, CARN, and CARN + UREA treatments (148 vs 95, 101, and 108 micromol/L, respectively). Intravenous L-carnitine administration influenced plasma glucose and NEFA concentrations in sheep and, when administered 30 min preceding an OULT, prevented the development of subclinical hyperammonemia in sheep. PMID- 9856406 TI - Experiential learning in the animal sciences: effect of 13 years of a beef cattle management practicum. AB - As the source of students shifts from rural to urban and suburban communities, students entering agricultural programs have less practical livestock experience. The career goals indicated by most of these students require knowledge of and experience with practical applications of their course work. The objective of this study was to examine the profile of students enrolled in an experiential beef cattle course 1) to describe the demographic and occupational characteristics of students enrolled and 2) to assess the perceived value of course activities to graduates completing the course as related to their skill attainment and career development. The questionnaire was sent to all 312 students who were enrolled in the course from 1983 to 1996. Over 61% of the respondents indicated they had enrolled in the course to gain experience working with beef cattle. Over 39% took the course to enhance their application to the College of Veterinary Medicine. When asked to rate the value of the course, as it related to skill development, they noted it was most helpful in teaching cattle handling skills, growth performance measurement, live animal evaluation, nutritional management, carcass and meat product value determination, and breed identification. PMID- 9856405 TI - Metabolic adaptation to experimentally increased glucose demand in ruminants. AB - Four Dorset wethers were studied in a Latin square design with 72-h periods to determine the metabolic adaptations that occur in support of increased glucose demand in ruminants. Wethers injected at 8-h intervals with excipient or a total of .5, 1.0, or 2.0 g/d of phlorizin excreted an average of 0, 72.7, 97.9, and 98.5 g/d of glucose into the urine, respectively. Both acute (2 to 24 h after the first injection) and chronic (8-h intervals from 8 to 72 h after the first injection) adaptations of plasma variables to phlorizin treatment were assessed. Concentrations of plasma glucose decreased linearly with increasing phlorizin dose during the 1st 24 h of treatment and tended to decrease linearly with phlorizin dose during 8 to 72 h of treatment. Urea N tended to increase linearly during 2 to 24 h and increased linearly during 8 to 72 h. Nonesterified fatty acids increased linearly with phlorizin injection during the entire treatment period. beta-Hydroxybutyrate increased quadratically with phlorizin injection during 2 to 24 h and tended to increase quadratically during 8 to 72 h. The ratio of insulin to glucagon tended to decrease linearly with phlorizin injection during the 1st 24 h but was unaffected from 8 to 72 h. Triiodothyronine, but not thyroxine, tended to decrease linearly with phlorizin injection during 8 to 72 h. Cortisol was not affected by treatment. Digestibilities of energy and N were not affected by treatment. Urinary energy excretion increased with phlorizin injection in proportion to the amounts of glucose excreted into the urine. These data indicate that phlorizin-treated wethers largely adapted to phlorizin treatment by 24 h after the first injection and are a suitable model for further investigations of hepatic adaptation to increased glucose demand in ruminants. PMID- 9856407 TI - Nucleotide sequence of ovine C/EBPepsilon gene. PMID- 9856408 TI - Polycystic ovary syndrome: What is it? Pathogenetic enigma and therapeutic dilemma. PMID- 9856409 TI - Polysystic ovary syndrome--loss of the apoptotic mechanism in the ovarian follicles? AB - Polycystic ovary syndrome (PCOS) is the most prevalent female endocrinopathy and the largest single cause of anovulatory infertility. The PCOS is characterized by multiple small antral follicles arrested in their development but nonatretic and viable. The hyperexpression of some growth factors (e.g. EGF/TGF alpha) in PCOS, considered to be survival or antiapoptotic factors, led to the hypothesis of their involvement in the blocking of apoptosis and atresia leading to an accumulation of multiple small antral follicles. Diminished FSH stimulation and accumulation of androgens could explain the arrest of progress to the preovulatory stage. Further investigation of the pathogenesis of PCOS is needed on the modulation of tumour suppressor and apoptosis genes such as p53, BAX or the APO/FAS system and the over expression of survival genes such as BCL2. PMID- 9856410 TI - Premature pubarche, ovarian hyperandrogenism, hyperinsulinism and the polycystic ovary syndrome: from a complex constellation to a simple sequence of prenatal onset. AB - Adolescent girls with a history of premature pubarche have an increased incidence of functional ovarian hyperandrogenism [a form of polycystic ovary syndrome (PCOS)] at adolescence, which is usually associated with hyperinsulinemia and dyslipemia. The hyperinsulinemia and lipid disturbances can often be detected in the prepubertal period and throughout puberty, and are associated with an exaggerated ovarian androgen synthesis. Birthweight SD scores are lower in premature pubarche girls than in control girls, and particularly so in those girls who show hyperinsulinemia and subsequently develop ovarian hyperandrogenism. Therefore, although the mechanisms interlinking the triad of premature pubarche, hyperinsulinism and ovarian hyperandrogenism remain enigmatic, these data indicate that the triad may result, at least in part, from a common early origin, rather than from a direct interrelationship later in life. PMID- 9856412 TI - Adrenal hyperandrogenism in the pathophysiology of polycystic ovary syndrome. PMID- 9856411 TI - The pathogenesis of polycystic ovary syndrome: lessons from ovarian stimulation studies. AB - In polycystic ovary syndrome (PCOS) the ovary produces markedly increased amounts of both androgens and estrogens in response to gonadotropin stimulation. Distinctive responses of 17-hydroxyprogesterone and androstenedione to ovarian stimulation testing suggest that ovarian hyperandrogenism is a result of dysregulation of theca cell androgen production which is intrinsic to the ovary. The occurrence of hyperestrogenism together with hyperandrogenism in PCOS suggests that whatever the abnormality of local regulatory factors of steroidogenesis, it affects granulosa as well as theca cells. Dysregulation is often associated with an increase in the number of follicles which evade atresia and reach the 2-8 mm stage of development. Autocrine/paracrine factors, especially those which are FSH-dependent, likely play an important role in the pathogenesis of the ovarian abnormality. Both LH and insulin hypersecretion probably play a secondary role in PCOS by amplifying the preexisting ovarian dysregulation. Because FSH secretion is under tight long-loop negative-feedback control and LH is not, hyperandrogenism is the primary clinical manifestation of dysregulation of steroid production in PCOS. However, anovulation in PCOS is most likely a result of excessive estrogen and inhibin production by multiple, small follicles which inhibit FSH secretory dynamics sufficiently to prevent selection of a dominant follicle. PMID- 9856413 TI - Polycystic ovary syndrome and insulin resistance: thrifty genes struggling with over-feeding and sedentary life style? AB - Almost two decades of research have greatly increased our knowledge in the complex field of metabolic aberrations in polycystic ovary syndrome, but still many problems remain unsolved. The statistical association between insulin levels and androgens originally put the focus on possible direct cause-and-effect relationships between these factors. Indeed there is evidence that insulin may affect ovarian functions in multiple ways, presumably in some cases causing anovulation and hyperandrogenism. Clearly, insulin may increase biologically active testosterone through reducing SHBG levels. Conversely, major increases in androgen levels may induce muscular changes leading to reduced insulin-mediated glucose uptake. There are suggestions of increased steroidogenesis in both ovarian and adrenal pathways, with the net result of increased androgen production. There are also findings supporting increased corticosteroid production, which could contribute to insulin resistance directly or through promoting accumulation of abdominal fat, a typical feature of over-weight women with PCOS. Free fatty acids, released in great amounts from abdominal fat, may induce insulin resistance. Insulin resistance may also be due to a primary aberration in the insulin receptor. Putatively increased serine phosphorylation may cause both impairment of the insulin signal and increased 17,20 lyase activity, thus suggesting a common cause for insulin resistance and increased androgen production. There are also findings supporting a high prevalence of beta cell dysfunction in PCOS, ranging from increased insulin secretion, not explained by insulin resistance or BMI, to failing beta-cell function, mainly in obese women during progress to glucose intolerance and NIDDM. Recent genetic findings also support a multifactorial genesis to PCOS, notably with positive findings both in genes regulating steroidogenesis and insulin secretion. It is suggested that PCOS is the result of "thrifty" genes, providing advantages in times of shortage of nutrition such as muscular strength, moderate abdominal fatness and decreased insulin sensitivity, i.e. an anabolic, energy saving constitution. However, when this constitution is exposed to unlimited food supplies and modern sedentary life style a full-blown PCOS with insulin resistance and infertility is triggered, presumably via several mechanisms, which follow a logical amplification system between two basic anabolic hormones, insulin and testosterone. PMID- 9856415 TI - How possible is the prevention of polycystic ovary syndrome development in adolescent patients with early onset of hyperandrogenism. AB - The polycystic ovary syndrome has a pubertal onset, with menstrual cycle irregularities and infertility in adulthood, as well as general health hazards such as increased risk for diabetes mellitus and myocardial infarction. Thus, prevention would be important. But as specific etiological factors are not known, nor is specific intervention. In order to use long-term intervention programs in children and adolescents, particularly rigid criteria must be used regarding proven safety and efficacy. In obese subjects, weight loss is very important, having beneficial effects on most aspects of the syndrome, such as subjective symptoms, infertility, hyperinsulinemia and related metabolic aberrations, and long-term health risks. Well controlled, long-term follow-up studies are needed on the role of prepubertal hyperandrogenism, hyperinsulinemia, gonadotropins, and ovarian morphological aspects in the development of PCOS, and possibilities of prevention. PMID- 9856416 TI - Estrogen-progestagen therapy in the management of the polycystic ovary syndrome. PMID- 9856414 TI - Insights into hypothalamic-pituitary dysfunction in polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is characterized by menstrual dysfunction and hyperandrogenism in the absence of other known causes. While the pathogenesis of PCOS remains elusive and is likely to involve abnormalities in several systems, there has long been an association of abnormal gonadotropin secretion with this disorder. In recent studies we have determined that 94% of women meeting the broad criteria for PCOS have an increased LH/FSH ratio. Several lines of evidence suggest that the mechanisms underlying the increased LH/FSH ratio in PCOS include an increased frequency of GnRH secretion. Decreased sensitivity to progesterone negative feedback on the GnRH pulse generator may play a role in this neuroendocrine defect. Additional factors which may contribute to the low to normal FSH levels in the face of increased LH include chronic mild estrogen increases and possibly inhibin. In addition to these effects on the differential control of FSH, there is increased pituitary sensitivity of LH secretion to GnRH. Both estrogen and androgens have been proposed as candidates mediating these effects. Superimposed on these underlying abnormalities in gonadotropin secretion is a marked inhibitory effect of obesity on LH secretion which may be mediated at either a pituitary or hypothalamic level. PMID- 9856417 TI - How actual is the treatment with antiandrogen alone in patients with polycystic ovary syndrome? PMID- 9856418 TI - The feasibility of long-term treatment of polycystic ovary syndrome with GnRH agonists. PMID- 9856419 TI - Attenuation of hyperinsulinemia in polycystic ovary syndrome: what are the options? PMID- 9856421 TI - Highlights 1995-1998. PMID- 9856420 TI - How actual is the dietary treatment in overweighting patients with polycystic ovary syndrome? PMID- 9856422 TI - New routes in the polycystic ovary syndrome labyrinth: a way out? PMID- 9856423 TI - A new form of Rickets during infancy: phosphate depletion-induced osteopenia due to antacid ingestion. PMID- 9856424 TI - Temperature, age, and recurrence of febrile seizure. AB - OBJECTIVE: Prediction of a recurrent febrile seizure during subsequent episodes of fever. DESIGN: Study of the data of the temperatures, seizure recurrences, and baseline patient characteristics that were collected at a randomized placebo controlled trial of ibuprofen syrup to prevent febrile seizure recurrences. SETTING: Two pediatric hospitals in the Netherlands. PATIENTS: A total of 230 children with an increased risk of febrile seizure recurrence. MAIN OUTCOME MEASURE: Seizure recurrence during a subsequent fever episode. RESULTS: A total of 509 episodes of fever were registered with 67 recurrences; 35 (52%) recurrences within the first 2 hours after fever of onset had a lower median temperature (39.3 degrees C) than 32 (48%) after more than 2 hours of fever (40.0 degrees C, P<.001). Poisson regression analysis resulted in 3 univariably significant (P<.05) predictors of a recurrence of seizure during a subsequent episode of fever. In a multivariable model, they were corrected for their correlation: interval between the last previous seizure and fever of onset less than 6 months (relative risk= 1.3 [95% confidence interval: 0.8-2.4]), age at fever of onset (relative risk=0.7 [95% confidence interval: 0.5-1.0] per year increase) and temperature at fever of onset (relative risk = 1.7 [95% confidence interval: 1.1-2.8] per degree Celsius increase). CONCLUSIONS: Half of the recurrent seizures occur in the first 2 hours after fever of onset of a subsequent fever episode. If seizure recurs at a later time, the temperature at seizure is higher compared with recurrences occurring in the first 2 hours of fever. Young age at fever of onset, high temperature at fever of onset, and high temperature during the episode of fever are associated with an increased risk of a recurrent febrile seizure at the moment that a child with a history of febrile seizures has fever again. PMID- 9856425 TI - Minor head injury in children: current management practices of pediatricians, emergency physicians, and family physicians. AB - OBJECTIVE: To describe variation in the clinical management of minor head trauma in children among primary care and emergency physicians. DESIGN: A survey of pediatricians, family physicians, and emergency physicians drawn from a random sample of members of the American Academy of Pediatrics, the American Academy of Family Physicians, and the appropriate American Medical Association specialty listings, respectively. Physicians were given clinical vignettes describing children presenting with normal physical examination results after minor head trauma. Different clinical scenarios (brief loss of consciousness or seizures) were also presented. Information was gathered on initial and subsequent management steps most commonly used by the physician. RESULTS: Surveys were returned by 765 (51%) of 1500 physicians. Of these, 303 (40%) were pediatricians, 269 (35%) family practitioners, and 193 (25%) emergency physicians. For minor head trauma without complications, observation at home was the most common initial physician management choice (n = 547, 72%). Observation in office or hospital was chosen by 81 physicians (11%). Head computed tomographic (CT) scan was chosen by 7 physicians (1%) and skull x-ray by 24 physicians (3%) as the first management option. Most physicians (n = 445, 80%) who initially chose observation at home would obtain a CT scan if the patient showed clinical deterioration. In the original scenario, if the patient had also sustained a loss of consciousness, 383 physicians (58%) altered management. Of these, 120 (18%) chose CT, 13 (2%) chose skull x-ray, 1 (1%) chose magnetic resonance imaging, 141 (21%) chose inpatient observation, and 125 (19%) chose a combination of CT scanning and observation. With seizures, 595 (90%) altered management, with 176 physicians (27%) choosing CT scan, 5 (1%) skull x-ray, 60 (9%) inpatient observation, and 299 (45%) a combination of radiological evaluation and observation. CONCLUSIONS: Most physicians surveyed chose clinic or home observation for initial management of minor pediatric head trauma. Clinical management was more varied when patients had sustained either loss of consciousness or seizures. Further study of the appropriate management of minor head trauma in children is needed to guide physicians in their care. PMID- 9856426 TI - Accidental and suicidal adolescent poisoning deaths in the United States, 1979 1994. AB - OBJECTIVE: To describe the epidemiological features of poisoning deaths in adolescents in the United States. DESIGN: Descriptive analysis of poisoning deaths in persons aged 10 to 19 years in the United States from January 1, 1979, to December 31, 1994, based on national mortality data. STUDY POPULATION: Adolescents whose cause of death was identified as poisoning using International Classification of Diseases, Ninth Revision codes. MAIN OUTCOME MEASURE: Nature of injury (accident vs suicide). RESULTS: There were 4129 suicides and 3807 accidental deaths due to poisoning. Victims were most frequently male and white. However, poisoning was more often the method of suicide in adolescent girls than in boys (28.0% vs 8.7%). The number of deaths (7138 vs 798) and death rate (2.36 vs 0.28 per 100,000 population) were higher in 15- to 19-year-olds vs 10- to 14 year-olds. The distribution of substances involved was different for 10- to 14 year-olds compared with 15- to 19-year-olds and for suicides compared with accidents. Among 10- to 14-year-olds, drugs other than alcohol accounted for 232 (85.3%) of 272 suicides but only 118 (22.4%) of 526 accidental deaths. Gases and vapors played an important role in accidental deaths and suicides in 15- to 19 year-olds and in accidents in 10- to 14-year-olds. CONCLUSIONS: The rates of suicides and accidental poisoning deaths were lower in 10- to 14-year-olds compared with 15- to 19-year-olds. Areas where injury-prevention efforts might have an influence on adolescent fatalities include management of depression, substance abuse education, and use of carbon monoxide detectors or shutoff switches. PMID- 9856427 TI - Decreased response to phototherapy for neonatal jaundice in breast-fed infants. AB - OBJECTIVE: To evaluate the efficacy of phototherapy for nonhemolytic hyperbilirubinemia in breast-fed and formula-fed infants and infants receiving formula and breast milk. DESIGN: Prospective study. SETTING: Nursery for healthy infants. METHOD: Full-term healthy infants with nonhemolytic hyperbilirubinemia (bilirubin concentration, >255 micromol/L [14.9 mg/dL] or 222 micromol/L [13.0 mg/dL] at ages younger than 48 hours) were treated with conventional phototherapy by using daylight fluorescent lamps. Three groups of infants were studied: group 1, formula-fed infants; group 2, breast-fed infants; and group 3, infants receiving formula and breast milk. All patterns of feeding started at birth. Phototherapy was terminated only when bilirubin concentrations had decreased to less than 185 micromol/L (10.8 mg/dL); the minimum exposure period was 24 hours. RESULTS: A total of 163 infants were studied: group 1, 79; group 2, 34; and group 3, 50. The age at the start of exposure was comparable in all groups. The mean+/ SD weight loss as a percentage of birth weight was as follows: group 1, 2.8%+/ 5.0%; group 2, 6.1%+/-3.4%; and group 3, 3.2%+/-2.6%. The duration of exposure to phototherapy was as follows: group 1, 54.1+/-20.8 hours; group 2, 64.6+/-25.1 hours; and group 3, 54.9+/-21.5 hours; the 24-hour rate of decrease in the bilirubin concentration was as follows: group 1, 18.6%+/-11.7%; group 2, 17.1%+/ 9.6%; and group 3, 22.9%+/-9.4%. The overall rate of decrease in the bilirubin concentration for the duration of exposure to phototherapy was as follows: group 1, 0.8%+/-0.3% per hour; group 2, 0.6%+/-0.3% per hour; and group 3, 0.8%+/-0.3% per hour. Weight loss at the start of phototherapy was significantly greater in group 2 compared with group 1 (P<.001) and group 3 (P<.001), although the hemoglobin and hematocrit values were comparable. The duration of exposure to phototherapy was not significantly different in the 3 groups (P=.06); however, the duration of exposure of group 2 infants was 10 hours more than that of the other 2 groups. The 24-hour rate of decrease in the bilirubin concentration in group 3 was significantly better than that of group 2 (P = .007) and group 3 (P = .02); the rates of decrease for groups 2 and 3 were similar (P = .52). The overall rate of decrease in the bilirubin concentration during the duration of exposure to phototherapy in group 2 was significantly less than that of group 1 (P = .002) and group 3 (P<.001); the rates for groups 1 and 3 were similar (P = .35). The postexposure rebound bilirubin concentrations were comparable in all groups during the first 2 days; however, the duration of moderate jaundice in group 2 was more prolonged. CONCLUSIONS: The response to phototherapy of group 2 infants was significantly slower than that of group 3 and group 1 infants; this response was still of adequate efficacy. The addition of formula to the feedings for totally breast-fed infants, without suspension of breast-feeding, would enhance the efficacy of phototherapy and reduce exposure time. PMID- 9856428 TI - Effect of prednisone on response to influenza virus vaccine in asthmatic children. AB - OBJECTIVE: To evaluate the immunogenicity of the influenza virus vaccine in children receiving short-course (a burst) prednisone therapy for acute asthmatic exacerbations. DESIGN: Prospective cohort study. SETTING: Outpatient pediatric clinic of a military medical center. PATIENTS: Children aged 6 months to 18 years requiring the 1996 influenza virus vaccine were eligible for the study. A total of 58 children were enrolled initially. The control group included 37 asthmatic children requiring less than 900 microg/d of inhaled prednisone and their siblings. The prednisone group included 21 children vaccinated at the beginning of a course of prednisone prescribed to treat an asthma exacerbation. Thirty-one control subjects (84%) and 19 patients in the prednisone group (90%) completed the study. Dropout was due to failure to come in for the postvaccination serum sampling. INTERVENTIONS: All study patients underwent immunization with the 1996 1997 trivalent subvirion influenza virus vaccine (FluShield; Wyeth Laboratories Inc, Marietta, Pa) containing 15-microg hemagglutinin antigens each of A/Texas/36/91 (H1N1) (A/H1), A/Wuhan/359/95 (H3N2)(A/H3), and B/Beijing/184/93 (B). The prednisone cohort received a burst of oral prednisone therapy (2 mg/kg per day for 5 days). MAIN OUTCOME MEASURES: To assess the immunogenicity of the vaccine between both groups, at least a 4-fold rise in titer and end titers of at least 1:40 to each of the 3 antigens were compared. Mean changes in geometric titers to the 3 antigens were also compared. RESULTS: Proportion of patients in each group with at least a 4-fold rise in titer to each of the influenza antigens was as follows: for A/H3N3 antigen, 15 patients (79%) in the prednisone group vs 22 controls (71%) (P = .74); for A/ H1N1 antigen, 16 patients in the prednisone group (84%) vs 20 controls (64%) (P = .20); and for B antigen, 7 patients in the prednisone group (37%) vs 8 controls (26%) (P = .53). Proportion of patients in each group with an end titer of at least 1:40 to each of the antigens was as follows: for A/ H3N2 antigen, 18 patients in the prednisone group (95%) vs 28 controls (90%) (P = .69); for A/H1N1 antigen, 17 patients in the prednisone group (89%) vs 26 controls (84%) (P = .99); and for B antigen, 7 patients in the prednisone group (37%) vs 13 controls (42%) (P = .99). There were also no significant differences between groups in the mean changes in geometric titers to any of the 3 antigens. CONCLUSIONS: Prednisone bursts did not diminish the response of asthmatic children to the 1996 influenza virus vaccine, compared with controls. Children can be effectively vaccinated against influenza virus while they are receiving prednisone therapy bursts for asthmatic exacerbations. PMID- 9856430 TI - Cost-effectiveness analysis of lead poisoning screening strategies following the 1997 guidelines of the Centers for Disease Control and Prevention. AB - OBJECTIVE: To compare blood lead (BPb) poisoning screening strategies in light of the 1997 recommendations by the Centers for Disease Control and Prevention, Atlanta, Ga. DESIGN: Cost-effectiveness analysis from the perspective of the health care system to compare the following 4 screening strategies: (1) universal screening of venous BPb levels; (2) universal screening of capillary BPb levels; (3) targeted screening of venous BPb levels for those at risk; and (4) targeted screening of capillary BPb levels for those at risk. Costs of follow-up testing and treatment were included in the model. RESULTS: Only universal venous screening detected all BPb levels of at least 0.48 micromol/L (10 microg/dL). Universal capillary screening detected between 93.2% and 95.5% of cases, depending on the prevalence of elevated BPb levels. Targeted screening was the least sensitive strategy for detecting cases. Venous testing identified between 77.3% and 77.9% of cases, and capillary testing detected between 72.7% and 72.8% of cases. In high-prevalence populations, universal venous screening minimized the cost per case ($490). In low- and medium-prevalence populations, targeted screening using venous testing minimized the cost per case ($729 and $556, respectively). In all populations, regardless of screening strategy, venous testing resulted in a lower cost per case than capillary testing. Sensitivity analyses of all parameters in this model demonstrated that this conclusion is robust. CONCLUSIONS: Universal screening detects all cases of lead poisoning and is the most cost-effective strategy in high-prevalence populations. In populations with lower prevalence, the cost per case detected using targeted screening is less than that of universal screening. The benefit of detecting a greater number of cases using universal screening must be weighed against the extra cost of screening. Regardless of whether a strategy of universal or targeted screening is used, the cost per case using venous testing is less than that of capillary testing. PMID- 9856429 TI - The relationship between asthma and obesity in urban minority children and adolescents. AB - OBJECTIVE: To investigate the relationship between asthma and obesity in children and adolescents. DESIGN: Medical record review. SETTING: Urban community health center. PARTICIPANTS: One hundred seventy-one children aged 4 to 16 years, 85 with asthma and 86 nonasthmatic controls. MAIN OUTCOME MEASURES: Diagnosis of asthma, age, and sex-adjusted body mass index (weight in kilograms divided by the square of the height in meters). RESULTS: Seventy-eight percent of the sample was Hispanic, 17% was African American, 2% were white, and 3% were other minorities. There were significantly more children with asthma (30.6%) who were very obese (> or =95th body mass index percentile) compared with controls (11.6%) (P=.004). Children with asthma were also significantly more overweight than controls (mean+/-SD, 22.5%+/-28.3% vs 12.0%+/-19.6% overweight; P=.004). The difference in obesity between children with asthma and controls was significant for both sexes and across the 4.5 to 10.9 years and 11 to 16 years age groups. Asthma severity was not related to obesity. CONCLUSION: Asthma is a risk factor for obesity in children and adolescents. PMID- 9856431 TI - Motor vehicle occupant deaths among Hispanic and black children and teenagers. AB - OBJECTIVE: To determine the risk of motor vehicle occupant deaths per unit of travel for Hispanic, non-Hispanic black, and non-Hispanic white children (aged 5 12 years) and teenagers (aged 13-19 years). DESIGN: Comparison of 1989 to 1993 motor vehicle occupant death rates of children and teenagers by race, ethnicity, and sex by using data on mortality from the National Center for Health Statistics, travel data from the 1990 Nationwide Personal Transportation Survey, and 1990 US census data. RESULTS: Among children 5 to 12 years old, race/ ethnicity differences per 100000 persons were unremarkable, but per billion vehicle-miles of travel, the rates were 14 for non-Hispanic blacks, 8 for Hispanics, and 5 for non-Hispanic whites. Among teenagers aged 13 to 19, the rates per 100000 persons were highest for non-Hispanic whites; however, the rates per billion vehicle-miles were 45 for Hispanics, 34 for non-Hispanic blacks, and 30 for non-Hispanic whites. Black and Hispanic male teenagers had substantially higher death rates per billion vehicle-miles of travel than either white male teenagers or female teenagers in any racial/ethnic group. CONCLUSIONS: Black and Hispanic children and teenagers are at higher risk of dying in motor vehicle crashes when they travel. Greater public health attention is needed to address these increased risks. PMID- 9856432 TI - What do parents know about lead poisoning? The Chicago Lead Knowledge Test. Pediatric Practice Research Group. AB - OBJECTIVES: To examine the extent of parental knowledge about lead poisoning and its prevention and to determine characteristics associated with accurate lead knowledge. SETTING: Twenty-three pediatric practices and 1 family practice in Chicago, Ill, and its suburbs. METHODS: A 24-question test regarding lead poisoning and its prevention (Chicago Lead Knowledge Test) was developed based on lead specialists' review and parental test-retest reliability. One point was assigned for each correct response. It was self-administered by a sample of 2225 parents of 0- to 6-year-old children visiting study practices. A 1-way analysis of variance (ANOVA) was used to determine the association of demographic descriptors with test scores. RESULTS: Respondents had a mean age of 33 years. Ninety percent were mothers, 49% were college graduates, and 80% were home owners. Fifteen percent lived in homes built before 1950, of which 36% were remodeled or renovated during the last 6 months. Respondents' youngest children were 80% white, 10% Hispanic, 5% African American, and 5% other. Ten percent received Medicaid and 86% had other medical insurance. Thirty-four percent recalled receipt of lead information from a health care provider, and 2.4% had had a child with a blood lead level of 0.48 micromol/L (10 microg/dL) or higher. The mean Chicago Lead Knowledge Test score was 12.2 (SD, 3.7). Questions related to lead exposure were more often answered correctly than those related to prevention and diet. In the ANOVA model, those who recalled receipt of lead information from a health care provider, college graduates, respondents aged 30 years or older, Hispanic respondents, and those living in homes built before 1950 had higher scores (all ANOVA P< or =.001). CONCLUSIONS: Parents do not have much knowledge of ways to prevent childhood lead poisoning. Information from a health care provider can aid parental knowledge. The Chicago Lead Knowledge Test is a new self-administered tool to help evaluate lead education programs. PMID- 9856433 TI - Evaluation of head injury in a pediatric emergency department: pretrauma and posttrauma system. AB - OBJECTIVE: To determine if trauma center protocols affect the number of tests and consultations performed and the length of time spent in the emergency department or hospital. DESIGN: A retrospective review and comparison of treatment for children with isolated head injury admitted to the emergency department before trauma center designation (group 1, 1985), and 5 years after implementation of trauma center protocols (group 2, 1991). SETTING: Urban children's hospital, level I trauma center. RESULTS: One hundred sixty-five children met the enrollment criteria in 1985 and 162 met the criteria in 1991. Falls were the predominant mechanism of injury (55%) for both years. For patients with moderate injury (Glasgow Coma Scale score, 9-12) or severe injury (Glasgow Coma Scale score, <9), there was no difference in radiographic or laboratory evaluation. For patients with minimal head injury (Glasgow Coma Scale score, 15, no loss of consciousness, amnesia, seizure, focal neurologic findings, or persistent symptoms) and minor head injury (Glasgow Coma Scale score, >12, and loss of consciousness or amnesia), more radiologic and laboratory studies were done in 1991 that showed no clinically significant abnormalities. Patients with minimal head injury in group 2 were 14 times more likely to have cranial computed tomographic scans performed (95% confidence interval [CI], 3.4-67); 11 times more likely to have cervical spine radiographs (95% CI, 2.2-76.6); and 23 times more likely to have hepatic enzymes obtained (95% CI, 3-491). These differences persisted when analyzed by both the age of the patient and mechanism of injury. CONCLUSIONS: Application of trauma system protocols to isolated head injury patient evaluation results in increased use of laboratory and radiologic services. These practices have the potential to increase the cost of medical care without significantly improving outcome. PMID- 9856434 TI - Growth of infants and young children born small or large for gestational age: findings from the Third National Health and Nutrition Examination Survey. AB - OBJECTIVES: To compare the growth profiles of infants and young children born small for gestational age (SGA, < 10th percentile birth weight for gestation) or large for gestational age (LGA, > or =90th percentile) with those appropriate for gestational age, and to document the expected growth patterns through early childhood based on national health examination survey data. SAMPLE: Infants and children, 2 to 47 months of age, who were born in the United States and examined using the Third National Health and Nutrition Examination Survey (1988-1994). MAIN OUTCOME MEASURES: Measurements of growth status based on normalized distributions (z scores or standard deviation units [SDUs] for weight, length, and head circumference. RESULTS: Prevalence rates were as follows: SGA infants, 8.6%; appropriate for gestational age infants, 80.9%; and LGA infants, 10.5%. Infants who were SGA appeared to catch up in weight in the first 6 months, but thereafter maintained a deficit of about -0.75 SDUs compared with infants who were appropriate for gestational age. The weight status of LGA infants remained at about +0.50 SDUs through 47 months of age. Length and head circumference were also associated with birth weight status, averaging over -0.60 SDUs for SGA infants and +0.43 SDUs for LGA infants. CONCLUSIONS: Birth weight status is related to growth rates in infancy and early childhood, which underscores the importance of considering child growth relative to birth status when using growth charts. Small for gestational age infants remain shorter and lighter and have smaller head circumferences, while LGA infants grow longer and heavier and have larger head circumferences. PMID- 9856435 TI - Performance of advanced resuscitation skills by pediatric housestaff. AB - OBJECTIVE: To describe pediatric housestaff resuscitation experience and their ability to perform key resuscitation skills. DESIGN: Cohort study of 63 pediatric residents in a university-based training program. PARTICIPANTS AND METHODS: Investigators observed, scored, and timed resident performance on 4 key resuscitation skills. Cognitive ability was tested with 4 written scenarios. Housestaff provided self-reports of the number of months since their last American Heart Association Pediatric Advanced Life Support course, number of mock and actual codes attended, number of times skills were performed, and self confidence with respect to resuscitation. RESULTS: A total of 45 pediatric residents (71%) participated. Median cognitive score was 5 (range, 1-5). Of all residents, 44 (97%) successfully bag mask-ventilated the mannequin; 24 (53%) and 36 (80%) used the correct bag and mask size, respectively. Thirty-nine residents (87%) placed a tube in the mannequin trachea, 12 (27%) checked that suction was working prior to intubation, and 30 (67%) chose the correct endotracheal tube size. Forty residents (89%) discharged the defibrillator, and 25 (56%) and 32 (71%) correctly chose asynchronous mode and infant paddles, respectively. Thirty eight residents (84%) inserted an intraosseous line; 35 (78%) had correct placement. Median times for successful skill completion were 83 seconds for bag mask ventilation, 136 seconds for intubation, 149 seconds for defibrillation, and 68 seconds for intraosseous line placement. CONCLUSION: Pediatric housestaff previously trained in pediatric advanced life support were generally able to reach the end point of 4 key resuscitation skills but less frequently performed the specific subcomponents of each skill. This poor performance and the prolonged time to skill completion suggest the need for greater attention to detail during training. PMID- 9856436 TI - Radiological case of the month. Epiglottic cyst. PMID- 9856437 TI - Picture of the month. Acrodermatitis enteropathica-like rash in a breast-fed, full-term infant with zinc deficiency. PMID- 9856438 TI - Pathological case of the month. Temporal triangular alopecia and aplasia cutis congenita. PMID- 9856439 TI - Rickets and secondary craniosynostosis associated with long-term antacid use in an infant. PMID- 9856440 TI - Ovarian cyst and torsion in a young infant. PMID- 9856441 TI - Advisability of colonoscopy in the management of ingested lead poisoning. PMID- 9856442 TI - The advantages and disadvantages of computer-aided instruction for pediatric residents. PMID- 9856443 TI - The possible link between erythropoietic protoporphyria and Alport syndrome. PMID- 9856444 TI - Waived testing in the physicians' office laboratory. PMID- 9856445 TI - Thinking globally, acting locally: AMPA receptor turnover and synaptic strength. PMID- 9856446 TI - Dissecting semaphorin signaling. PMID- 9856447 TI - Making proteins at the synapse: activity-regulated translation and CPEB. PMID- 9856448 TI - Crystal clear structure/function relationships for GluRs. PMID- 9856449 TI - A lie detector test for presynaptic capacitance measurements. PMID- 9856450 TI - Specifying motor neurons and their connections. PMID- 9856451 TI - Immunologic complexity in neurons. PMID- 9856452 TI - Inside and outside the focus of attention. PMID- 9856453 TI - Tau mutations cause frontotemporal dementias. PMID- 9856454 TI - Allosteric receptors after 30 years. PMID- 9856455 TI - Neuroethology: a meeting of brain and behavior. PMID- 9856456 TI - Wrapper, a novel member of the Ig superfamily, is expressed by midline glia and is required for them to ensheath commissural axons in Drosophila. AB - The midline glia are specialized, nonneuronal cells at the midline of the Drosophila central nervous system (CNS). During development, the midline glia provide guidance cues for extending axons. At the same time, they migrate and help separate the two axon commissures. They then wrap around and ensheath the commissural axons. In many segments, a few of the glia do not enwrap the axons, and these cells die. The wrapper gene encodes a novel member of the immunoglobulin (Ig) superfamily. Wrapper protein is expressed specifically on the surface of midline glia. In wrapper mutant embryos, the midline glia express their normal guidance cues and migrate normally. However, they do not ensheath the commissural axons, and as a result, the glia die. In the absence of Wrapper, the two axon commissures are not properly separated. PMID- 9856457 TI - Growth arrest failure, G1 restriction point override, and S phase death of sensory precursor cells in the absence of neurotrophin-3. AB - More than half of the dorsal root ganglion (DRG) neurons are lost by excessive cell death coinciding with precursor proliferation and cell cycle exit in neurotrophin-3 null mutant (NT-3-/-) mice. We find that in the absence of NT-3, sensory precursor cells fail to arrest the cell cycle, override the G1 phase restriction point, and die by apoptosis in S phase, which can be prevented in vivo by a cell cycle blocker. Uncoordinated cell cycle reentry is preceded by a failure of nuclear N-myc downregulation and is paralleled by the activation of the full repertoire of G1 and S phase cell cycle proteins required for cell cycle entry. Our results provide evidence for novel activity of neurotrophins in cell cycle control and point toward an N-myc sensitization to cell death in the nervous system that is under the control of NT-3. PMID- 9856458 TI - Identification and characterization of novel substrates of Trk receptors in developing neurons. AB - Neurotrophins influence growth and survival of specific populations of neurons through activation of Trks, members of the receptor tyrosine kinase (RTK) family. In this report, we describe the identification and characterization of two substrates of Trk kinases, rAPS and SH2-B, which are closely related Src homolog 2 (SH2) domain-containing signaling molecules. rAPS and SH2-B are substrates of TrkB and TrkC in cortical neurons and SH2-B is a substrate of TrkA in sympathetic neurons. Moreover, rAPS and SH2-B bind to Grb2, and both are sufficient to mediate NGF induction of Ras, MAP kinase (MAPK), and morphological differentiation of PC12 cells. Lastly, antibody perturbation and transient transfection experiments indicate that SH2-B, or a closely related molecule, is necessary for NGF-dependent signaling in neonatal sympathetic neurons. Together, these observations indicate that rAPS and SH2-B mediate Trk signaling in developing neurons. PMID- 9856459 TI - Pax6 controls radial glia differentiation in the cerebral cortex. AB - Radial glia cells perform a dual function in the developing nervous system as precursor cells and guides for migrating neurons. We show here that during forebrain neurogenesis, the transcription factor Pax6 is specifically localized in radial glia cells of the cortex but not of the basal telencephalon. In Pax6 deficient mice, cortical radial glia cells were altered in their morphology, number, tenascin-C (TN-C) expression, and cell cycle. We show that some of these alterations are cell-autonomous, whereas others were rescued by coculturing with wild-type cortical cells. Our results suggest that Pax6 plays an essential role in the differentiation of cortical radial glia. Thus, despite their widespread distribution, radial glia cells are regionally specified in the developing CNS. PMID- 9856460 TI - Developmental regulation of neuronal K+ channels by target-derived TGF beta in vivo and in vitro. AB - The functional expression of Ca2+-activated K+ channels (KCa) in developing chick ciliary ganglion (CG) neurons requires interactions with target tissues and preganglionic innervation. Here, we show that the stimulatory effects of target tissues are mediated by an isoform of TGFbeta. Exposure of cultured CG neurons to TGFbeta1, but not TGFbeta2 or TGFbeta3, caused robust stimulation of KCa. The KCa stimulatory effects of target tissue extracts were blocked by a neutralizing pan TGFbeta antiserum but not by specific TGFbeta2 or TGFbeta3 antisera. Intraocular injection of TGFbeta1 caused robust stimulation of KCa, whereas intraocular injection of pan-TGFbeta antiserum inhibited expression of KCa in CG neurons developing in vivo. The effects of TGFbeta1 were potentiated by beta-neuregulin 1, a differentiation factor expressed in preganglionic neurons. PMID- 9856461 TI - Functionally distinct NMDA receptors mediate horizontal connectivity within layer 4 of mouse barrel cortex. AB - In sensory areas of neocortex, thalamocortical afferents project primarily onto the spiny stellate neurons of Layer 4. Anatomical evidence indicates that these cells receive most of their excitatory input from other cortical neurons, including other spiny stellate cells. Although this local network must play an important role in sensory processing, little is known about the properties of the neurons and synapses involved. We have produced a slice preparation of mouse barrel cortex that isolates Layer 4. We report that excitatory interaction between spiny stellate neurons is largely via N-methyl-D-aspartate receptors (NMDARs) and that a given neuron contains more than one type of NMDAR, as distinguished by voltage dependence. Thus, spiny stellate cells act as effective integrators of powerful and persistent NMDAR-mediated recurrent excitation. PMID- 9856462 TI - Activity-dependent modulation of synaptic AMPA receptor accumulation. AB - Both theoretical and experimental work have suggested that central neurons compensate for changes in excitatory synaptic input in order to maintain a relatively constant output. We report here that inhibition of excitatory synaptic transmission in cultured spinal neurons leads to an increase in mEPSC amplitudes, accompanied by an equivalent increase in the accumulation of AMPA receptors at synapses. Conversely, increasing excitatory synaptic activity leads to a decrease in synaptic AMPA receptors and a decline in mEPSC amplitude. The time course of this synaptic remodeling is slow, similar to the metabolic half-life of neuronal AMPA receptors. Moreover, inhibiting excitatory synaptic transmission significantly prolongs the half-life of the AMPA receptor subunit GluR1, suggesting that synaptic activity modulates the size of the mEPSC by regulating the turnover of postsynaptic AMPA receptors. PMID- 9856463 TI - Neuropilin-2 is a receptor for semaphorin IV: insight into the structural basis of receptor function and specificity. AB - Neuropilins bind secreted members of the semaphorin family of proteins. Neuropilin-1 is a receptor for Sema III. Here, we show that neuropilin-2 is a receptor for the secreted semaphorin Sema IV and acts selectively to mediate repulsive guidance events in discrete populations of neurons. neuropilin-2 and semaIV are expressed in strikingly complementary patterns during neurodevelopment. The extracellular complement-binding (CUB) and coagulation factor domains of neuropilin-2 confer specificity to the Sema IV repulsive response, and these domains of neuropilin-1 are necessary and sufficient for binding of the Sema III semaphorin (sema) domain. The coagulation factor domains alone are necessary and sufficient for binding of the Sema III immunoglobulin- (Ig-) basic domain and the unrelated ligand, vascular endothelial growth factor (VEGF). Lastly, neuropilin-1 can homomultimerize and form heteromultimers with neuropilin-2. These results provide insight into how interactions between neuropilins and secreted semaphorins function to coordinate repulsive axon guidance during neurodevelopment. PMID- 9856464 TI - Neuropilin-1 extracellular domains mediate semaphorin D/III-induced growth cone collapse. AB - Somatosensory axon outgrowth is repulsed when soluble semaphorin D (semD) binds to growth cone neuropilin-1 (Npn-1). Here, semD ligand binding studies of Npn-1 mutants demonstrate that the sema domain binds to the amino-terminal quarter, or complement-binding (CUB) domain, of Npn-1. By herpes simplex virus- (HSV-) mediated expression of Npn-1 mutants in chick retinal ganglion cells, we show that semD-induced growth cone collapse requires two segments of the ectodomain of Npn-1, the CUB domain and the juxtamembrane portion, or MAM (meprin, A5, mu) domain. In contrast, the transmembrane segment and cytoplasmic tail of Npn-1 are not required for biologic activity. These data imply that the CUB and MAM ectodomains of Npn-1 interact with another transmembrane growth cone protein that in turn transduces a semD signal into axon repulsion. PMID- 9856465 TI - Mammalian circadian autoregulatory loop: a timeless ortholog and mPer1 interact and negatively regulate CLOCK-BMAL1-induced transcription. AB - We report the cloning and mapping of mouse (mTim) and human (hTIM) orthologs of the Drosophila timeless (dtim) gene. The mammalian Tim genes are widely expressed in a variety of tissues; however, unlike Drosophila, mTim mRNA levels do not oscillate in the suprachiasmatic nucleus (SCN) or retina. Importantly, hTIM interacts with the Drosophila PERIOD (dPER) protein as well as the mouse PER1 and PER2 proteins in vitro. In Drosophila (S2) cells, hTIM and dPER interact and translocate into the nucleus. Finally, hTIM and mPER1 specifically inhibit CLOCK BMAL1-induced transactivation of the mPer1 promoter. Taken together, these results demonstrate that mTim and hTIM are mammalian orthologs of timeless and provide a framework for a basic circadian autoregulatory loop in mammals. PMID- 9856466 TI - Molecular analysis of mammalian timeless. AB - We cloned the mouse cDNA of a mammalian homolog of the Drosophila timeless (tim) gene and designated it mTim. The mTim protein shows five homologous regions with Drosophila TIM. mTim is weakly expressed in the suprachiasmatic nuclei (SCN) but exhibits robust expression in the hypophyseal pars tuberalis (PT). mTim RNA levels do not oscillate in the SCN nor are they acutely altered by light exposure during subjective night. mTim RNA is expressed at low levels in several peripheral tissues, including eyes, and is heavily expressed in spleen and testis. Yeast two-hybrid assays revealed an array of interactions between the various mPER proteins but no mPER-mTIM interactions. The data suggest that PER PER interactions have replaced the function of PER-TIM dimers in the molecular workings of the mammalian circadian clock. PMID- 9856467 TI - Coordinated interactions between hippocampal ripples and cortical spindles during slow-wave sleep. AB - Sleep is characterized by a structured combination of neuronal oscillations. In the hippocampus, slow-wave sleep (SWS) is marked by high-frequency network oscillations (approximately 200 Hz "ripples"), whereas neocortical SWS activity is organized into low-frequency delta (1-4 Hz) and spindle (7-14 Hz) oscillations. While these types of hippocampal and cortical oscillations have been studied extensively in isolation, the relationships between them remain unknown. Here, we demonstrate the existence of temporal correlations between hippocampal ripples and cortical spindles that are also reflected in the correlated activity of single neurons within these brain structures. Spindle ripple episodes may thus constitute an important mechanism of cortico-hippocampal communication during sleep. This coactivation of hippocampal and neocortical pathways may be important for the process of memory consolidation, during which memories are gradually translated from short-term hippocampal to longer-term neocortical stores. PMID- 9856468 TI - CPEB-mediated cytoplasmic polyadenylation and the regulation of experience dependent translation of alpha-CaMKII mRNA at synapses. AB - Long-term changes in synaptic efficacy may require the regulated translation of dendritic mRNAs. While the basis of such regulation is unknown, it seemed possible that some features of translational control in development could be recapitulated in neurons. Polyadenylation-induced translation of oocyte mRNAs requires the cis-acting CPE sequence and the CPE-binding protein CPEB. CPEB is also present in the dendritic layers of the hippocampus, at synapses in cultured neurons, and in postsynaptic densities of adult brain. alpha-CaMKII mRNA, which is localized in dendrites and is necessary for synaptic plasticity and LTP, contains two CPEs. These CPEs are bound by CPEB and mediate polyadenylation induced translation in injected Xenopus oocytes. In the intact brain, visual experience induces alpha-CaMKII mRNA polyadenylation and translation, suggesting that this process likely occurs at synapses. PMID- 9856469 TI - Mechanism of action of rab3A in mossy fiber LTP. AB - In mossy fiber synapses of the hippocampal CA3 region, LTP is induced by cAMP and requires the synaptic vesicle protein rab3A. In contrast, CA1-region synapses do not exhibit this type of LTP. We now show that cAMP enhances glutamate release from CA3 but not CA1 synaptosomes by (1) increasing the readily releasable pool as tested by hypertonic sucrose; (2) potentiating release evoked by KCl depolarization, which opens voltage-gated Ca2+ channels; and (3) by enhancing Ca2+ action on the secretory apparatus as monitored by the Ca2+-ionophore ionomycin. In rab3A-deficient synaptosomes, forskolin still enhances KCl- and sucrose-induced glutamate release but not ionomycin-induced release. Our results show that cAMP has multiple actions in mossy fiber synapses, of which only the direct activation of the secretory apparatus requires rab3A and functions in mfLTP. PMID- 9856470 TI - NMDA induces long-term synaptic depression and dephosphorylation of the GluR1 subunit of AMPA receptors in hippocampus. AB - Brief bath application of N-methyl-D-aspartate (NMDA) to hippocampal slices produces long-term synaptic depression (LTD) in CA1 that is (1) sensitive to postnatal age, (2) saturable, (3) induced postsynaptically, (4) reversible, and (5) not associated with a change in paired pulse facilitation. Chemically induced LTD (Chem-LTD) and homosynaptic LTD are mutually occluding, suggesting a common expression mechanism. Using phosphorylation site-specific antibodies, we found that induction of chem-LTD produces a persistent dephosphorylation of the GluR1 subunit of AMPA receptors at serine 845, a cAMP-dependent protein kinase (PKA) substrate, but not at serine 831, a substrate of protein kinase C (PKC) and calcium/calmodulin-dependent protein kinase II (CaMKII). These results suggest that dephosphorylation of AMPA receptors is an expression mechanism for LTD and indicate an unexpected role of PKA in the postsynaptic modulation of excitatory synaptic transmission. PMID- 9856471 TI - Involvement of a postsynaptic protein kinase A substrate in the expression of homosynaptic long-term depression. AB - Hippocampal N-methyl-D-aspartate (NMDA) receptor-dependent long-term synaptic depression (LTD) is associated with a persistent dephosphorylation of the GluR1 subunit of AMPA receptors at a site (Ser-845) phosphorylated by cAMP-dependent protein kinase (PKA). In the present study, we show that dephosphorylation of a postsynaptic PKA substrate may be crucial for LTD expression. PKA activators inhibited both AMPA receptor dephosphorylation and LTD. Injection of a cAMP analog into postsynaptic neurons prevented LTD induction and reversed previously established homosynaptic LTD without affecting baseline synaptic transmission. Moreover, infusing a PKA inhibitor into postsynaptic cells produced synaptic depression that occluded homosynaptic LTD. These findings suggest that dephosphorylation of a PKA site on AMPA receptors may be one mechanism for NMDA receptor-dependent homosynaptic LTD expression. PMID- 9856472 TI - Submillisecond kinetics of glutamate release from a sensory synapse. AB - Exocytosis-mediated glutamate release from ribbon-type synaptic terminals of retinal bipolar cells was studied using AMPA receptors and simultaneous membrane capacitance measurements. Release onset (delay <0.8 ms) and offset were closely tied to Ca2+ channel opening and closing. Asynchronous release was not copious and we estimate that there are approximately 5 Ca2+ channels per docked synaptic vesicle. Depending on Ca2+ current amplitude, release occurred in a single fast bout or in two successive bouts with fast and slow onset kinetics. The second, slower bout may reflect a mobilization rate of reserve vesicles toward fusion sites that is accelerated by increasing Ca2+ influx. Bipolar cell synaptic ribbons thus are remarkably versatile signal transducers, capable of transmitting rapidly changing sensory input, as well as sustained stimuli, due to their large pool of releasable vesicles. PMID- 9856473 TI - Dendritic sodium spikes are variable triggers of axonal action potentials in hippocampal CA1 pyramidal neurons. AB - Several early studies suggested that spikes can be generated in the dendrites of CA1 pyramidal neurons, but their functional significance and the conditions under which they occur remain poorly understood. Here, we provide direct evidence from simultaneous dendritic and somatic patch-pipette recordings that excitatory synaptic inputs can elicit dendritic sodium spikes prior to axonal action potential initiation in hippocampal CA1 pyramidal neurons. Both the probability and amplitude of dendritic spikes depended on the previous synaptic and firing history of the cell. Moreover, some dendritic spikes occurred in the absence of somatic action potentials, indicating that their propagation to the soma and axon is unreliable. We show that dendritic spikes contribute a variable depolarization that summates with the synaptic potential and can act as a trigger for action potential initiation in the axon. PMID- 9856474 TI - G protein alpha subunit G alpha z couples neurotransmitter receptors to ion channels in sympathetic neurons. AB - The functional roles subserved by G(alpha)z, a G protein alpha subunit found predominantly in neuronal tissues, have remained largely undefined. Here, we report that G(alpha)z coupled neurotransmitter receptors to N-type Ca2+ channels when transiently overexpressed in rat sympathetic neurons. The G(alpha)z-mediated inhibition was voltage dependent and PTX insensitive. Recovery from G(alpha)z mediated inhibition was extremely slow but accelerated by coexpression with RGS proteins. G(alpha)z selectively interacted with a subset of receptors that ordinarily couple to N-type Ca2+ channels via PTX-sensitive Go/i proteins. In addition, G(alpha)z rescued the activation of heterologously expressed GIRK channels in PTX-treated neurons. These results suggest that G(alpha)z is capable of coupling receptors to ion channels and might underlie PTX-insensitive ion channel modulation observed in neurons under physiological and pathological conditions. PMID- 9856475 TI - Effects of PS1 deficiency on membrane protein trafficking in neurons. AB - We have examined the trafficking and metabolism of the beta-amyloid precursor protein (APP), an APP homolog (APLP1), and TrkB in neurons that lack PS1. We report that PS1-deficient neurons fail to secrete Abeta, and that the rate of appearance of soluble APP derivatives in the conditioned medium is increased. Remarkably, carboxyl-terminal fragments (CTFs) derived from APP and APLP1 accumulate in PS1-deficient neurons. Hence, PS1 plays a role in promoting intramembrane cleavage and/or degradation of membrane-bound CTFs. Moreover, the maturation of TrkB and BDNF-inducible TrkB autophosphorylation is severely compromised in neurons lacking PS1. We conclude that PS1 plays an essential role in modulating trafficking and metabolism of a selected set of membrane and secretory proteins in neurons. PMID- 9856477 TI - Alternative splicing of exons 29 and 30 in the neurofibromatosis type 1 gene. AB - Alternative splicing of exons 29 and 30 of the human neurofibromatosis type 1 (NF1) gene was detected by reverse transcription/polymerase chain reaction (RT PCR). Three different isoforms that omitted either one or both exons were identified (ex29-, ex30-, and ex29/30-). The alternatively spliced transcripts exhibited tissue-specific differences, with the ex30- variant apparent only in brain. All three isoforms altered the reading frame and introduced a stop codon in the adjacent downstream exon. Alternative splicing of this region of the NF1 gene also was detected in RNA from rats, although only the ex30- variant was observed. RNA from mice revealed only constitutive expression in this region of the NF1 gene. This study adds a new site of alternative processing to the complex expression of NF1. PMID- 9856476 TI - A novel mutation at a probable heme-binding ligand in neutrophil cytochrome b558 in atypical X-linked chronic granulomatous disease. AB - A membrane-bound cytochrome b558, a heterodimer consisting of gp91-phox and p22 phox, is a critical component of the superoxide (O2-)-generating reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in phagocytes. Chronic granulomatous disease (CGD) is characterized by recurrent bacterial infection caused by a defect of the oxidase. Both subunits are absent from phagocytes in typical X-linked recessive CGD patients who are primarily defective in gp91-phox. We report here an atypical case of X-linked CGD in which neutrophils showed a complete absence of O2--forming NADPH oxidase activity, but a small amount (about 10% of control) of both subunits was detected by immunoblot analysis. Spectrophotometric studies of the neutrophils with a recently developed sensitive method gave no evidence for the heme spectrum in the cytochrome b558, of this CGD. Reverse transcription/polymerase chain reaction and sequence analysis revealed a C to T transition replacing histidine at amino acid position 101 (His101) by tyrosine in gp91-phox. These results provide evidence that His101 of gp91-phox is the one of the heme-binding ligands of cytochrome b558. PMID- 9856478 TI - Identification and characterization of a novel cyclic nucleotide phosphodiesterase gene (PDE9A) that maps to 21q22.3: alternative splicing of mRNA transcripts, genomic structure and sequence. AB - Cyclic nucleotide-specific phosphodiesterases (PDEs) play an essential role in signal transduction by regulating the intracellular concentration of second messengers (cAMP and cGMP). We have identified and made an initial characterization of a full-length cDNA encoding a novel human cyclic nucleotide phosphodiesterase, PDE9A. At least four different mRNA transcripts (PDE9A1, A2, A3, A4) are produced as a result of alternative splicing of 5' exons, potentially changing the N-terminal amino acid sequences of the encoded proteins. All these predicted proteins would contain a 3',5'-cyclic nucleotide phosphodiesterase signature motif (Prosite no. PDOC00116). Northern blot analysis revealed several mRNA species of approximately 2.4 kb with varying expression patterns and intensities in most tissues examined, except blood. We have also isolated the mouse homolog of the human PDE9A2 mRNA transcript, pde9A2. The human and mouse isoforms have 93 and 83% sequence identity at the amino acid and nucleotide levels, respectively. PDE9A was mapped to 21q22.3, between TFF1 and D21S360. Comparison of the PDE9A1 cDNA with the genomic sequence from the region revealed that the gene is split into 20 exons that extend over 122 kb. Comparison of the physical map of the region and the genomic sequence further refines the mapping, with D21S113 being derived from intron 15. Several genetic disorders map to 21q22.3, including one form of bipolar affective disorder. Since functional disturbances in intraneuronal signal transmission via second messengers play an important role in the pathophysiology of affective disorders, PDE9A is a strong candidate for such a role by position and function. PMID- 9856479 TI - Functional defects of Cx26 resulting from a heterozygous missense mutation in a family with dominant deaf-mutism and palmoplantar keratoderma. AB - Mutations in GJB2 encoding the gap junction protein connexin-26 (Cx26) have been established as the basis of autosomal recessive non-syndromic hearing loss. The involvement of GJB2 in autosomal dominant deafness has also been proposed, although the putative mutation identified in one family with both deafness and palmoplantar keratoderma has recently been suggested to be merely a non-disease associated polymorphism. We have observed a similar phenotype in an Egyptian family that segregated with a heterozygous missense mutation of GJB2, leading to a non-conservative amino acid substitution (R75W). The deleterious dominant negative effect of R75W on gap channel function was subsequently demonstrated in the paired oocyte expression system. Not only was R75W alone incapable of inducing electrical conductance between adjacent cells, but it almost completely suppressed the activity of co-expressed wildtype protein. The Cx26 mutant W77R, which has been implicated in autosomal recessive deafness, also failed to form functional gap channels by itself but did not significantly interfere with the function of wildtype Cx26. These data provide compelling evidence for the serious functional consequences of Cx26 mutations in dominant and recessive deafness. PMID- 9856480 TI - Congenital atrichia in five Arab Palestinian families resulting from a deletion mutation in the human hairless gene. AB - Congenital atrichia is a rare autosomal recessive disorder of hair development, characterized by complete loss of hair shortly after birth. Evidence of linkage to chromosome 8p12 has been established, implicating the human homolog of the mouse hairless (hr) gene as a candidate gene. We have previously identified missense mutations in families with congenital atrichia. Here, we report the first deletion mutation (2147del C) in exon 9 of the human hairless gene leading to a frameshift and downstream premature termination codon in five Palestinian families of Arab origin. PMID- 9856481 TI - In vivo somatic mutations in Werner's syndrome. AB - The frequencies of mutant erythrocytes with loss of heterozygosity at the glycophorin A (GPA) locus and mutant CD4+ T cells lacking surface expression of the T-cell receptor alphabeta (TCR)/CD3 complex were measured by flow cytometry for Japanese Werner's syndrome (WRN) patients. The hemizygous and homozygous GPA mutant frequencies (GPA Mfs) and the TCR/CD3-defective mutant frequency (TCR Mf) in WRN patients were found to be significantly higher than those in normal controls in the same age range. However, because these Mfs in the patients are only about twice those in controls, it is difficult to conclude that the WRN gene mutations cause instability of somatic genes. This contrasts markedly with Bloom's syndrome (BLM) patients, whose GPA and TCR Mfs were previously reported to increase about 50- and 15-fold, respectively. The difference in Mfs is one aspect of the large variation in the phenotype observed between WRN and BLM patients, suggesting a different role of the responsible genes, both of which belong to the RecQ DNA helicase gene family, in the control of somatic mutagenesis. PMID- 9856482 TI - Chromosomal localization and genomic organization of the human retinal rod Na Ca+K exchanger. AB - The retinal rod Na-Ca+K exchanger is a unique calcium extrusion protein found only in the outer segments of retinal rod photoreceptors. Rod Na-Ca+K exchanger cDNA (NCKX1) has been cloned from bovine and human retinas. Here, we have used fluorescent in situ hybridization and radiation hybrid mapping to localize the human NCKX1 gene to chromosome 15q22. We have determined the genomic organization of human rod NCKX1 and found one intron in the 5' untranslated region and eight introns within the coding region. PMID- 9856483 TI - Selective complement C1s deficiency caused by homozygous four-base deletion in the C1s gene. AB - The complement system plays an important role in defense mechanisms by promoting the adherence of microorganisms to phagocytic cells and lysis of foreign organisms. Deficiencies of the first complement components, C1r/C1s, often cause systemic lupus erythematosus-like syndromes and severe pyogenic infections. Up to now no genetic analysis of the C1r/C1s deficiencies has been carried out. In the present work, we report the first genetic analysis of selective C1s deficiency, the patient having a normal amount of C1r. C1s RNA with a normal size was detected in patient's subcutaneous fibroblasts (YKF) by RNA blot analysis and RT PCR. The amount of C1s RNA was approximately one-tenth of the RNA from the human chondrosarcoma cell line, HCS2/8. In contrast, the levels of C1r and beta-actin RNA of YKF were similar to that of HCS2/8. Sequence analysis of C1s cDNA revealed a deletion at nucleotides 1087-1090 (TTTG), creating a stop codon (TGA) at position 94 downstream of the mutation site. Direct sequencing of the gene between the primers designed on intron 9 and exon 10 indicated the presence of the deletion on exon 10 of the gene. Quantitative Southern blot hybridization suggested the mutation was homozygous. The 4-bp deletion on exon 10 was also found in the patient's heterozygous mother who had normal hemolytic activity. PMID- 9856484 TI - Drug-selected complete restoration of superoxide generation in Epstein-Barr virus transformed B cells from p47phox-deficient chronic granulomatous disease patients by using a bicistronic retrovirus vector encoding a human multi-drug resistance gene (MDR1) and the p47phox gene. AB - Chronic granulomatous disease (CGD) is a group of disorders characterized by the failure of phagocytes to produce superoxide. One-third of the cases of CGD in the USA and Europe results from defects in the gene encoding p47phox, a cytoplasmic component of NADPH oxidase for superoxide generation. In this study, we constructed the bicistronic retrovirus vector Ha-MDR-IRES-p47, which carries cDNAs for a human multi-drug-resistance gene (MDR1) and p47phox. The amphotropic retroviral producer cells were generated, and the supernatant of the producer cells was used to transduce Epstein-Barr virus-transformed B (EBV-B) cells, established from B cells of p47phox-deficient CGD patients, as an in vitro model of gene therapy for p47phox-deficient CGD. The transduced cells expressed both P glycoprotein and p47phox protein, and the expression levels were increased after appropriate vincristine selection. The levels of superoxide production in the vincristine-selected cells were increased to a level similar to normal EBV-B cells. This result suggests that it is possible to achieve 100% correction of the CGD defect in p47phox-deficient EBV-B cells by using the bicistronic vector. This strategy could be employed not only in vitro, but also in vivo, in the gene therapy of a number of inherited diseases. PMID- 9856485 TI - Deletions in the Parkin gene and genetic heterogeneity in a Greek family with early onset Parkinson's disease. AB - Parkinson's disease is the second most common neurodegenerative disease after Alzheimer's disease and is manifested as a movement disorder. A positive family history is the second most important risk factor for developing the illness, after age. Both autosomal dominant and recessive forms of the illness have been described. Recently deletions in a novel gene, parkin, have been associated with the autosomal recessive form of the illness in Japanese families. In this study, we demonstrate that deletions of exons 5, 6 and 7 of the parkin gene are present in two affected individuals of a Greek pedigree with early onset Parkinson's disease. However, no deletions were identified in a different branch of the same pedigree with three affected individuals. These results suggest that deletions in the parkin gene will be found in other families besides those of Japanese origin and that there must be at least one additional locus responsible for early onset autosomal recessive Parkinson's disease. PMID- 9856486 TI - A common mutation in Sardinian autoimmune polyendocrinopathy-candidiasis ectodermal dystrophy patients. AB - Autoimmune polyendocrinopathy-candidiasisectodermal dystrophy (APECED; also called APS-1,) is a rare autosomal recessive disorder that is more frequent in certain isolated populations. It is characterized by two of the three major clinical symptoms that may be present: Addison's disease, and/or hypoparathyroidism and/or chronic mucocutaneous candidiasis. We have recently identified the gene for APECED, which we termed AIRE (for autoimmune regulator). AIRE is expressed in thymus, lymph nodes and fetal liver, and encodes a protein with two putative zinc fingers and other motifs suggestive of a transcriptional regulator. Seven mutations have been described to date, including R257X, the predominant Finnish and northern Italian APECED allele, which has also been observed in other patients of diverse origin on different haplotypes. A 13-bp deletion (1094-1106del) has also been observed in several patients of different geo-ethnic origin. The other described mutations appear to be rare. We present mutational analyses of the AIRE gene in ten Sardinian APECED families and show that there is a mutation, R139X, associated with one predominant haplotype unique to the Sardinian patients (18/20 independent alleles). The carrier frequency of R139X in Sardinia is 1.7%, giving an estimated population frequency of APECED of 1/14,400. Using linkage disequilibrium data, the estimated age of the R139X mutation is between 20 and 25 generations. A previously described 13-bp deletion was also observed on an allele of one patient. The identification of a single common Sardinian APECED mutation will facilitate its genetic diagnosis. Given the carrier frequency of R139X in the Sardinian population, AIRE may be implicated in the pathogenesis of other autoimmune diseases in the Sardinian population, particularly those affecting the endocrine system. PMID- 9856487 TI - The polymorphic 43Thr bcl-2 protein confers relative resistance to autoimmunity: an analytical evaluation. AB - We have found a novel polymorphic (Ala43Thr; ACC-->GCC) bcl-2 allele in a Japanese population. An in vitro expression study with a mouse IL-7-dependent pre B cell line has revealed that inhibition of the programmed cell death function of 43Thr bcl-2 protein is suppressed compared with that of normal 43Ala bcl-2 protein. Since bcl-2 expression in B-lymphoid cells elicits autoimmune disease in mice, we have investigated the possibility of whether a bcl-2 polymorphism has a different susceptibility to autoimmune disease. To evaluate the clinical impact of this polymorphism, the frequency of bcl-2 polymorphism was investigated in 221 children with insulin-dependent diabetes mellitus (IDDM), 237 adults with autoimmune disease (105 with rheumatoid arthritis, 57 with systemic lupus erythematosus, 55 with Sjogren's syndrome, and 20 others), and 290 healthy Japanese children and adults. The frequency of the 43Thr bcl-2 allele, either homozygous or heterozygous, was 14.5% in normal controls, 6.8% (P<0.01) in children with IDDM, and 8.0% (P<0.025) in adults with autoimmune disease. These results suggest that the 43Thr allele of bcl-2 confers resistance to autoimmune disease. The different anti-apoptotic function resulting from the different expression of bcl-2 protein in lymphocytes seems to be associated with the development of autoimmune disease, indicating that the bcl-2 gene affects human autoimmune disease. PMID- 9856488 TI - Pigmentary mosaicism in hypomelanosis of Ito. Further evidence for functional disomy of Xp. AB - We report on a female with mental and motor retardation, facial dysmorphism, abnormal pigmentation reminiscent to hypomelanosis of Ito (HI), and karyotypic mosaicism involving a small supernumerary marker chromosome. The marker chromosome was defined by fluorescence in situ hybridisation (FISH) as a ring X chromosome with breakpoints in the juxtacentromeric region. FISH analysis showed that the ring does not include the XIST locus at the X-inactivation centre and, therefore, may not be subject to X inactivation. X-inactivation studies with the HUMARA (human androgen receptor) and FMR1 assay showed a skewed X-inactivation pattern (85:15) with preferential inactivation of the paternal X chromosome. These results are discussed with respect to the role of functional disomy of Xp in the pathogenesis of HI. PMID- 9856489 TI - Molecular basis of neurological dysfunction coupled with haemolytic anaemia in human glucose-6-phosphate isomerase (GPI) deficiency. AB - Glucose-6-phosphate isomerase (GPI) deficiency, an autosomal recessive genetic disorder with the typical manifestation of nonspherocytic haemolytic anaemia, can be associated in some cases with neurological impairment. GPI has been found to be identical to neuroleukin (NLK), which has neurotrophic and lymphokine properties. To focus on the possible effects of GPI mutations on the central nervous system through an effect on neuroleukin activity, we analysed DNA isolated from two patients with severe GPI deficiency, one of them with additional neurological deficits, and their families. The neurologically affected patient (GPI Homburg) is compound heterozygous for a 59 A-->C (H20P) and a 1016 T ->C (L339P) exchange. Owing to the insertion of proline, the H20P and L339P mutations are likely to affect the folding and activity of the enzyme. In the second family studied, the two affected siblings showed no neurological symptoms. The identified mutations are 1166 A-->G (H389R) and 1549 C-->G (L517V), which are located at the subunit interface. We propose that mutations that lead to incorrect folding destroy both catalytic (GPI) and neurotrophic (NLK) activities, thereby leading to the observed clinical symptoms (GPI Homburg). Those alterations at the active site, however, that allow correct folding retain the neurotrophic properties of the molecule (GPI Calden). PMID- 9856490 TI - Refined mapping of the gene for thiamine-responsive megaloblastic anemia syndrome and evidence for genetic homogeneity. AB - Thiamine-responsive megaloblastic anemia (TRMA, also known as Rogers syndrome, OMIM 249270) is a rare autosomal recessive disorder characterized by a triad of megaloblastic anemia, diabetes mellitus, and sensorineural deafness. Patients respond, to varying degrees, to treatment with megadoses of thiamine. We have recently shown genetic linkage of the TRMA gene to a 16-centimorgan (cM) region on 1q23.2-1q23.3 based on the analysis of four large, inbred families of Alaskan, Italian, and Israeli-Arab origin. Here we narrow the TRMA interval down to 4 cM based on genetic recombination, homozygosity mapping, and linkage disequilibrium (highest LOD score of 12.5 at D1S2799, at a recombination fraction of 0). We provide further evidence that the TRMA gene is located in this region and confirm the homogeneity of the disease. In this analysis, we genotyped seven additional families of diverse ethnic origin (Pakistani, Indian, Italian, Brazilian, and Japanese), and analyzed additional markers in two previously reported families showing evidence of linkage disequilibrium in a large area of their haplotypes. The multi-system manifestations of TRMA suggest that thiamine has a pivotal role in a multiplicity of physiological processes. Mapping the TRMA gene and understanding the molecular basis of the disease might, thus, shed light on the role of thiamine in common disorders such as deafness, anemia, and diabetes. PMID- 9856491 TI - Two mutations remote from an exon/intron junction in the beta-hexosaminidase beta subunit gene affect 3'-splice site selection and cause Sandhoff disease. AB - Four unrelated Japanese patients with infantile Sandhoff disease (beta hexosaminidase beta-subunit deficiency) have been studied for the molecular basis of their severe phenotype. Two patients had complex base substitutions; one patient was homoallelic for a triple mutation (P417L, K121R, and S255R) and the other was a compound heterozygote of a double (P417L and K121R) mutation and the triple mutation. K121R is known to be a functional polymorphism, while P417L (exon 11, +8 C-->T) generates predominantly an abnormally spliced mRNA at base +112 of exon 11 and has been described in two patients with a juvenile form of the disease. The mild phenotype is attributed to the presence of a small amount of normally spliced mRNA. S255R is a novel mutation without prior description in the literature. An expression study of the normally spliced cDNA with the double and the triple mutations gave about 70% and 30% of normal activity, respectively. This finding suggests that S255R further reduces the catalytic activity of the already below-threshold amount of normally spliced mRNA and accounts for the more severe phenotype in our patients. In the other two patients, a novel disease causing base transition was found within intron 10, away from the intron/exon junction (-17 a-->g). This mutation caused abnormal 3' splicing at position -37 of intron 10, and no normally spliced product was detectable upon RT-PCR analysis. We noted an unusually low splice site score (61.8) for the exon 10/intron 11 junction and suspected that this might be partially responsible for the aberrant splicing in these mutations. To test this hypothesis, we constructed four chimeric cDNAs all with an additional intron 10 inserted and evaluated their splicing efficiency. They, respectively, had the normal sequence, P417L (exon 11, +8 C-->T), the intronic mutation (-17 a-->g), and the intronic mutation with an artificially engineered intron 10/exon 11 junction of a higher splice site score (85.1). Of the total transcripts, 67% and 32% were correctly spliced in the normal chimeric construct and P417L, respectively, while no normally spliced product was generated either in the chimeric construct with -17 a-->g or in that with a high splice site score. The sequence around the adenosine -17 residue upstream of the normal acceptor splice site in this report, UGCAAU (-21 to -16), matches the consensus branchpoint sequence YNYRAY (Y, pyrimidine; R, purine; N, any base) reported in the literature. The mutation in this study is most likely to abolish lariat formation because the artificial site of the high splice site score did not improve splicing efficiency. PMID- 9856492 TI - Evidence of an increased risk of hearing loss in heterozygous carriers in a Wolfram syndrome family. AB - Wolfram syndrome (MIM 222300) is characterized by juvenile-onset diabetes mellitus and optic atrophy. Previous linkage analyses in the United States and UK families have indicated that the gene for Wolfram syndrome (WFS) is localized on the short arm of chromosome 4. We herein confirm the linkage of the WFS locus to D4S3023 on 4p with a two-point LOD score of 3.42 in a large Japanese family with Wolfram syndrome. Multipoint linkage analysis revealed the maximum LOD score of 4.82 between D4S3023 and D4S394. We also evaluated putative health risks in carriers by multiple logistic analysis with independent variables, age, gender, and numbers of affected haplotypes and with dependent variables, such as hearing loss, diabetes mellitus, polyuria, incontinence, psychological illness, and visual acuity. The results showed that the putative disease haplotype increased a risk of hearing loss (odds ratio =35.68, 95% confidence interval =4.12-308.95) and diabetes mellitus (odds ratio =7.57, 95% confidence interval =2.03-28.23) independently. This is the first report of an increased health risk of illness in carriers, other than for psychiatric disease. PMID- 9856493 TI - Familial clustering of rheumatoid arthritis with other autoimmune diseases. AB - Previous studies have shown that rheumatoid arthritis aggregates within families. However, no formal genetic analysis of rheumatoid arthritis in pedigrees together with other autoimmune diseases has been reported. We hypothesized that there are genetic factors in common in rheumatoid arthritis and other autoimmune diseases. Results of odds-ratio regression and complex segregation analysis in a sample of 43 Caucasian pedigrees ascertained through a rheumatoid arthritis proband or matched control proband, revealed a very strong genetic influence on the occurrence of both rheumatoid arthritis and other autoimmune diseases. In an analysis of rheumatoid arthritis alone, only one inter-class measure, parent sibling, resulted in positive evidence of aggregation. However, three inter-class measures (parent-sibling, sibling-offspring, and parent-offspring pairs) showed significant evidence of familial aggregation with odds-ratio regression analysis of rheumatoid arthritis together with all other autoimmune diseases. Segregation analysis of rheumatoid arthritis alone revealed that the mixed model, including both polygenic and major gene components, was the most parsimonious. Similarly, segregation analysis of rheumatoid arthritis together with other autoimmune diseases revealed that a mixed model fitted the data significantly better than either major gene or polygenic models. These results were consistent with a previous study which concluded that several genes, including one with a major effect, is responsible for rheumatoid arthritis in families. Our data showed that this conclusion also held when the phenotype was defined as rheumatoid arthritis and/or other autoimmune diseases, suggesting that several major autoimmune diseases result from pleiotropic effects of a single major gene on a polygenic background. PMID- 9856494 TI - Elucidation of the exon-intron structure and size of the human protein kinase C beta gene (PRKCB). AB - As part of a transcriptional mapping project on human chromosome 16p12, a genomic contig was constructed that spanned the alternatively spliced human protein kinase C beta gene (PRKCB). PRKCB was determined to consist of 18 exons covering approximately 375 kb, with a particularly large intron of over 150 kb between exons 2 and 3. PRKCB is nearly 19 times larger than the highly homologous Drosophila melanogaster protein kinase C gene (dPKC), which has a similar-sized open reading frame but only 13 exons. This increase in size has occurred mostly as a result of expansion of introns, with intron size in the human gene averaging 22 kb compared with 1.5 kb in dPKC. The difference in gene size correlates with the difference in genome size, with the human haploid genome being nearly 18 times larger than the 170 Mb Drosophila haploid genome. PMID- 9856495 TI - Examination of the X chromosome by STS-PCR screening for the presence of submicroscopic deletions. AB - Cytogenetically undetectable deletions are suspected to be an important cause of mental retardation and developmental delay, as suggested by the observation that about 7% of children with undiagnosed mental retardation have rearrangements affecting the chromosome ends. Screening the whole genome for regions of aneuploidy smaller than 5 Mb is not feasible, but the availability of a high resolution map of the X chromosome means that it is possible to look for deletions in males by PCR. We have screened 96 affected males and their 96 unaffected fathers with 110 markers distributed across the X chromosome. No deletions were found in either group. Our results show that the prevalence of deletions greater than 1 Mb in children with mental retardation is less than 3.9% (95% confidence interval). We conclude that X chromosome deletions in the size range 1-5 Mb are a rare cause of mental retardation in males. PMID- 9856496 TI - Genomic structure of the human glucose 6-phosphate translocase gene and novel mutations in the gene of a Japanese patient with glycogen storage disease type Ib. AB - Glycogen storage disease (GSD) type Ib is an autosomal recessive disorder caused by a deficiency in microsomal glucose 6-phosphate (G6P) translocase. A gene mutated in GSD type Ib patients has recently been isolated. We have determined the entire sequence of the human G6P translocase gene by PCR-directed sequencing. The gene spans approximately 5 kb of genomic DNA and contains eight exons. Analysis of DNA from a Japanese patient with GSD type Ib revealed new compound heterozygous mutations; a T to C transition at cDNA position 521 resulting in W118R, and an A to C transversion at the -2 splicing acceptor site of intron 1. Reverse transcription (RT)-PCR from leukocyte RNA of the patient revealed the abnormally spliced transcript. These results further support the suggestion that the gene is causative for GSD Ib and should be useful in the molecular diagnosis of such patients. PMID- 9856497 TI - Noonan-like phenotype in monozygotic twins with a duplication-deficiency of the long arm of chromosome 18 resulting from a maternal paracentric inversion. AB - We report on newborn monozygotic twins with a Noonan-like phenotype, and multiple congenital anomalies due to a monocentric recombinant chromosome 18. The mother carried a paracentric inversion of the long arm of chromosome 18, inv(18)(q21.1q22.3). Cytogenetic, fluorescent in situ hybridization, comparative genomic hybridization and DNA marker analyses allowed the delineation of the deleted (18q22.3-qter) and duplicated (18q12.1-q21.1) chromosomal regions in the recombinant chromosome 18, and suggest that this duplication-deletion chromosome 18 resulted from breakage of a dicentric recombinant chromosome 18 with subsequent reconstitution of telomeric sequences on the long arm. Marked variability is observed in the phenotypic expression of the same chromosomal anomaly in these monozygotic twins. The clinical findings of these patients are compared with those reported in proximal 18q-duplication and distal 18q-deletion patients. The clinical features of both infants are compatible with Noonan syndrome, suggesting that a locus for this syndrome may be located on the long arm of chromosome 18. PMID- 9856498 TI - The molecular basis of C6 deficiency in the western Cape, South Africa. AB - Deficiency of the sixth component of human complement (C6) has been reported in a number of families from the western Cape, South Africa. Meningococcal disease is endemic in the Cape and almost all pedigrees of total C6 deficiency (C6Q0) have been ascertained because of recurrent disease. We have sequenced the expressed exons of the C6 gene from selected cases and have found three molecular defects leading to total deficiency: 879delG, which is the common defect in the Cape and hitherto unreported, and 1195delC and 1936delG, which have been previously reported in African-Americans. We also show that the 879delG and 1195delC defects are associated with characteristic C6/C7 region DNA marker haplotypes, although small variations were observed. The 1936delG defect was observed only once in the Cape, but its associated haplotype could be deduced. The data from the haplotypes indicate that these three molecular defects account for the defects in all the 38 unrelated C6Q0 individuals we have studied from the Cape. We have also observed the 879delG defect in two Dutch C6-deficient kindreds, but the 879delG defect in the Cape probably did not come from The Netherlands. PMID- 9856499 TI - Complement C7 deficiency: seven further molecular defects and their associated marker haplotypes. AB - Seven further molecular bases of C7 deficiency are described. All these new molecular defects involve single-nucleotide events, deletions and substitutions, some of which alter splice sites, and others codons. They are distributed along the C7 gene, but predominantly towards the 3' end. All were found in compound heterozygous individuals. The C6/C7 marker haplotypes associated with most C7 defects are tabulated. PMID- 9856500 TI - Screening with the FMR1 protein test among mentally retarded males. AB - The fragile X syndrome is characterized by X-linked mental retardation with additional features such as a long face with large protruding ears, macroorchidism, and eye-gaze avoidance. The disorder is caused by an abnormally expanded CGG repeat within the first exon of the fragile X mental retardation (FMR1) gene that is associated with shutdown of transcription and absence of the fragile X mental retardation protein (FMRP). Detection of patients and carriers of the fragile X syndrome is done by DNA analysis of the CGG repeat, whereas the FMRP antibody test allows rapid detection of male patients using bloodsmears. In a screening program for the fragile X syndrome in the southwest of the Netherlands, 412 males with mental retardation of unknown cause were subjected to the protein test. The patients were scored for fragile X features and their DNA tested for the FMR1 mutation, as reported previously. The FMRP test detected two fragile X patients with a repeat expansion in FMR1, whereas normal protein expression was observed in all the retarded male patients with a normal repeat. The FMRP test was found to be suitable for screening among a large population of retarded males. The results also suggest that mutations other than the CGG repeat leading to absence of detectable FMRP are apparently rare among mentally retarded males. PMID- 9856501 TI - Chromosomal mapping of three human LAMMER protein-kinase-encoding genes. AB - The eukaryotic LAMMER protein kinase family is encoded by at least three loci in the human genome, designated CLK1, 2, and 3. We have mapped these loci to 2q33, 1q21, and 15q24, respectively, by fluorescent in situ hybridization. Additionally, a CLK2 pseudo-gene has been located to 7p15-21. PMID- 9856502 TI - Physical mapping of the nail patella syndrome interval at 9q34: ordering of STSs and ESTs. PMID- 9856503 TI - Functional analysis of the C(-188)A polymorphism of the human leptin promoter. AB - Mutational analysis of the promoter region of the leptin gene in morbidly obese Finnish subjects had revealed a previously unidentified C(-188)A polymorphism in the proximal promoter that showed a weak association with elevated serum leptin levels in obese male carriers of the variant (-188A) allele. In this study we demonstrated that neither expression of reporter gene constructs driven by wild type (-188C) or variant (-188A) proximal promoter regions, nor assay of binding of cellular proteins reveal a genotype-related difference in promoter activity. PMID- 9856504 TI - Functional characterisation of mutations in the ligand-binding domain of the androgen receptor gene in patients with androgen insensitivity syndrome. AB - Five mutations in the ligand-binding domain of the androgen receptor gene were identified in patients with complete (A765T, C784Y, R831X and M895T) or partial (R840G) androgen insensitivity. A765T and R831X have been reported previously whereas the other three mutations are novel. Receptors carrying these mutations were transiently expressed in COS-1 cells, and androgen binding and capacity to transactivate an androgen-responsive reporter gene were assayed. C784Y led to abolished androgen binding and transactivating capacity, R840G and M895T showed reduced specific binding and partial transactivation. The in vitro functions of the R840G and M895T mutants were improved with supraphysiological concentrations of steroid. PMID- 9856505 TI - Cell therapy and tissue engineering for renal replacement therapy. PMID- 9856506 TI - The NKF research and training program: a report to its alumni and supporters and a promise for the future. National Kidney Foundation. PMID- 9856507 TI - Regulation of renal urate excretion: a critical review. AB - Uric acid metabolism is reviewed as it relates mainly to kidney and electrolyte disorders, with emphasis on the difficulties in understanding urate transport because of its bidirectional transport and the species differences in which animal data may not have relevance to the human condition. A critical review of the effects of pyrazinamide and extracellular volume expansion on urate transport raises questions about the current popular teachings that pyrazinamide exclusively blocks tubule urate secretion and extracellular volume expansion has a major role in controlling urate excretion. There appears to be a renal salt wasting syndrome with overlapping clinical features that make it indistinguishable from the syndrome of inappropriate secretion of antidiuretic hormone (SIADH), except possibly for extracellular volume depletion. Hypouricemia and the elevation in the fractional excretion of urate (%E/Furate) are extensively reviewed with a proposal to use the persistence of hypouricemia and elevated %E/Furate after the correction of hyponatremia to differentiate these patients from those with SIADH. An algorithm is proposed to differentiate one group from the other. A plasma natriuretic factor has been shown in some with probable renal salt wasting, which includes patients with AIDS, cancer, and pulmonary and intracranial diseases. The natriuretic factor may have etiologic implications and diagnostic and therapeutic applications. PMID- 9856508 TI - Genetic testing for renal diseases: medical and ethical considerations. AB - Advances in understanding the genetic basis of renal disorders will soon allow for the clinical use of genetic diagnostic testing. In this article, we review renal diseases with a known genetic basis and the current methods available for genetic testing. We then examine the potential medical indications for genetic testing, with special attention to autosomal dominant polycystic kidney disease (ADPKD). Because clinicians will be faced with patients considering genetic testing, we review the ethical considerations regarding genetic testing for renal diseases, recent genetic privacy legislation, and the special role genetic testing may have in transplantation. We conclude with a review of the necessary elements of informed consent, which provides the ethical foundation for patients deciding about genetic testing with the assistance of their physicians. PMID- 9856509 TI - Morphological studies of mesothelial cells in CAPD effluent and their clinical significance. AB - We studied the mesothelial cells in the effluent of patients undergoing continuous ambulatory peritoneal dialysis (CAPD) and their relationship with CAPD duration, peritoneal function, peritoneal sclerosis (PS), and sclerosing encapsulating peritonitis (SEP) in 49 patients (26 men, 23 women) treated for 3 to 161 months with CAPD. Three patients had SEP and five patients had PS. The overnight effluent was drained and centrifuged. The cell differentiation and surface area of mesothelial cells were studied by a computed light microscope system after cytospin preparation staining. The surface area of 50 cells was measured. The mesothelial cells were classified into three types according to their morphological appearance: normal cell type, with a mean surface area of 335.6+/-31.0 microm2 and mean nucleocytoplasmic ratio (N/C ratio) of 0.66+/-0.01; dyskaryotic cell type, with a mean surface area of 570.5+/-35.9 microm2 and N/C ratio 0.58+/-0.06; and giant cell type, with a mean surface area of 1,821.0+/ 68.8 microm2 and N/C ratio of 0.06+/-0.01. There was a low but significant correlation between the fast peritoneal equilibration test and surface area (r=0.495; P=0.0120) and a highly significant correlation between CAPD duration and mean surface area (r=0.719; P < 0.0001). This increased cell surface area was because of both an increased surface area of normal and dyskaryotic cells and an increase in the number of dyskaryotic and giant cells. The mean surface area in patients with SEP was 709.3+/-125.4 microm2 and that in patients with PS was 586.6+/-55.2 microm2. Giant cells were found in the effluent of all three patients with SEP and three of the patients with PS. In conclusion, a marked correlation was found between the surface area of effluent mesothelial cells and the duration of CAPD. Giant cells were almost always found in the effluent of patients with SEP and PS. The surface area of mesothelial cells in the effluent might reflect morphological changes in the peritoneum during CAPD. These morphological changes and the measurement of the size of mesothelial cells may predict critical derangement of peritoneal membrane. PMID- 9856510 TI - Effect of hemoperfusion with polymyxin B-immobilized fiber on plasma endothelin-1 and endothelin-1 mRNA in monocytes from patients with sepsis. AB - Hemoperfusion using polymyxin B-immobilized fiber (PMX-F) is reported to be an effective treatment for sepsis. The aim of the present study is to assess whether plasma endothelin-1 (ET-1) and ET-1 messenger RNA (mRNA) levels in peripheral blood monocytes are altered in patients with sepsis and whether PMX-F treatment affects plasma ET-1 and monocyte ET-1 mRNA levels. Sixteen patients with sepsis and 20 healthy volunteers were included in this study. Plasma ET-1 concentration was measured by radioimmunoassay (RIA), and plasma levels of transforming growth factor-beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha), and interleukin 1beta (IL-1beta) were measured by enzyme-linked immunosorbent assay (ELISA). Sixteen patients with sepsis were treated with direct hemoperfusion using PMX-F columns. Blood endotoxin levels decreased significantly from 35 to 10 pg/mL after two treatments of direct hemoperfusion, each for 2 hours. Patients with sepsis showed significantly increased levels of plasma ET-1 (P < 0.001) and monocyte ET 1 mRNA (P < 0.001) compared with healthy volunteers. However, no differences in plasma levels of TGF-beta, TNF-alpha, and IL-1beta existed between patients with sepsis and healthy volunteers. Increased plasma ET-1 levels and monocyte ET-1 mRNA levels in patients with sepsis decreased significantly after PMX-F treatment (P < 0.01). These data suggest that the secretion of ET-1 from peripheral-blood monocytes may be stimulated by endotoxin, and PMX-F treatment may be effective in reducing ET-1 secretion in patients with sepsis. PMID- 9856511 TI - Origin and phenotypic features of hyperplastic epithelial cells in collapsing glomerulopathy. AB - The characteristic pathological feature of collapsing glomerulopathy (CG) is marked cell hyperplasia and hypertrophy within the glomeruli. The present study investigated the phenotypic alteration of hyperplastic epithelial cells in CG to determine their origin. Renal biopsy specimens from two patients with CG were analyzed by immunohistochemical staining, using markers for podocytes (PHM-5), parietal epithelial cells (PECs; cytokeratin), and cell proliferation (Ki-67). In collapsed glomeruli, hyperplastic and hypertrophic epithelial cells were frequently connected to PECs and collapsed glomerular basement membranes (GBMs). These epithelial cells were more often Ki-67 positive and expressed cytokeratin, whereas PHM-5 was almost invariably negative. Serial section analysis showed that a small number of hyperplastic epithelial cells expressed both PHM-5 and cytokeratin, suggesting phenotypic conversion between podocytes and PECs. Moreover, cytokeratin-positive cells were associated with the sclerotic glomerular segments. Thus, we suggest that the majority of hyperplastic and hypertrophic epithelial cells in CG are of PEC origin. These epithelial features may participate in the development of characteristic tuft collapse and glomerulosclerosis in CG. PMID- 9856512 TI - Left ventricular mass and diastolic function in normotensive young adults with autosomal dominant polycystic kidney disease. AB - Left ventricular hypertrophy is often found very early in the course of autosomal dominant polycystic kidney disease (ADPKD). Diastolic dysfunction has been shown in hypertensive adult patients with ADPKD with increased left ventricular mass (LVM), but there are no data about diastolic function in the young ADPKD population without hypertension and with normal renal function. To evaluate very early alterations in cardiac structure and diastolic function in young normotensive patients with ADPKD, color Doppler echocardiography was performed in 46 young normotensive patients with ADPKD and 35 healthy subjects. LVM, transmitral pulsed Doppler flow (diastolic function), and valvular abnormalities were studied. Patients with ADPKD showed higher LVM indices (LVMIs) than controls (89.7+/-17.3 v 68.5+/-17.2 g/m2; P < 0.0001). Peak early diastolic velocity (E wave) deceleration time and isovolumic relaxation time were significantly prolonged in patients with ADPKD compared with controls (E wave deceleration time, 182.5+/-51.3 v 149.4+/-34 msec; P=0.002; isovolumic relaxation time, 97.7+/ 17.5 v 79+/-15 msec; P=0.0001). No differences were found in valvular abnormalities in the two groups. In conclusion, young normotensive patients with ADPKD showed increased LVMIs and Doppler abnormalities consistent with early diastolic dysfunction. PMID- 9856513 TI - Cystic fibrosis and the phenotypic expression of autosomal dominant polycystic kidney disease. AB - Recent experiments in cultured cyst epithelial cells from kidneys of patients with autosomal dominant polycystic kidney disease (ADPKD) have shown that the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) is present in the apical surface of these cells and mediates chloride (Cl-) and fluid secretion in vitro. To determine whether the presence of CF with the expression of mutated CFTR proteins modifies cyst formation in ADPKD, we studied a large family with both inherited diseases. ADPKD in this family is linked to PKD1. The family is composed of 26 members; 11 members with ADPKD, 4 members with CF, and 2 members with both diseases. Renal volumes measured by computerized tomography (CT), calculated creatinine clearances, and other clinical parameters in the family members with ADPKD and CF were compared with those in the family members with ADPKD alone, as well as to a large population of patients with ADPKD. The patients with CF and ADPKD, but not the CF heterozygote carriers with ADPKD, had less severe polycystic kidney and liver disease, as indicated by normal renal function; smaller renal volume, even when corrected for height and body surface area; and the absence of hypertension and liver cysts. These observations suggest that the coexistence of CF may reduce the severity of ADPKD. PMID- 9856514 TI - Posttransplantation acute tubular necrosis: risk factors and implications for graft survival. AB - Previous studies aimed at identifying the causes, risk factors, and outcome of kidney transplant recipients with delayed graft function (DGF) have yielded controversial results. We retrospectively analyzed the causes and risk factors for DGF in 263 cadaveric kidney transplantations from November 1988 to March 1997 in one center. Causes of DGF were assessed by postoperative graft evolution and graft biopsy. Univariate and multivariate analysis were used to investigate the risk factors for DGF induced by acute tubular necrosis (ATN). Seventy-six patients (29%) had DGF, which was caused by ATN in 70 patients (92.1%) and acute rejection (AR) in 6 patients (7.9%). Therefore, we focused on risk factors and consequences for ATN-induced DGF. In monofactorial analysis, ATN was significantly associated with greater weight and presence of an atheromatous disease in both donor and recipient. Other risk factors for ATN were older age of donor, recipient American Society of Anesthesiology (ASA) physical status category IV, cold ischemia time (CIT), and transplantation using the right kidney. The multivariate analysis showed that donor and recipient weight, donor age, transplantation using the right kidney, preservation in Eurocollins solution, ASA score, and CIT were associated with ATN. The incidence of rejection and renal function were not different at 3 months or 1 and 5 years. ATN is the main cause of DGF in kidney transplant recipients. ATN is caused by donor and recipient vascular background, grafting the right kidney, and CIT. ATN does not appear to have an adverse effect on long-term kidney function. PMID- 9856515 TI - Serum creatinine levels in the US population: third National Health and Nutrition Examination Survey. AB - This report describes the distribution of serum creatinine levels by sex, age, and ethnic group in a representative sample of the US population. Serum creatinine level was evaluated in the third National Health and Nutrition Examination Survey (NHANES III) in 18,723 participants aged 12 years and older who were examined between 1988 and 1994. Differences in mean serum creatinine levels were compared for subgroups defined by sex, age, and ethnicity (non Hispanic white, non-Hispanic black, and Mexican-American). The mean serum creatinine value was 0.96 mg/dL for women in the United States and 1.16 mg/dL for men. Overall mean creatinine levels were highest in non-Hispanic blacks (women, 1.01 mg/dL; men, 1.25 mg/dL), lower in non-Hispanic whites (women, 0.97 mg/dL; men, 1.16 mg/dL), and lowest in Mexican-Americans (women, 0.86 mg/dL; men, 1.07 mg/dL). Mean serum creatinine levels increased with age among both men and women in all three ethnic groups, with total US mean levels ranging from 0.88 to 1.10 mg/dL in women and 1.00 to 1.29 mg/dL in men. The highest mean creatinine level was seen in non-Hispanic black men aged 60+ years. In the total US population, creatinine levels of 1.5 mg/dL or greater were seen in 9.74% of men and 1.78% of women. Overall, among the US noninstitutionalized population, 10.9 million people are estimated to have creatinine values of 1.5 mg/dL or greater, 3.0 million have values of 1.7 mg/dL or greater, and 0.8 million have serum creatinine levels of 2.0 mg/dL or greater. Mean serum creatinine values are higher in men, non Hispanic blacks, and older persons and are lower in Mexican-Americans. In the absence of information on glomerular filtration rate (GFR) or lean body mass, it is not clear to what extent the variability by sex, ethnicity, and age reflects normal physiological differences rather than the presence of kidney disease. Until this information is known, the use of a single cutpoint to define elevated serum creatinine values may be misleading. PMID- 9856516 TI - Preferential expression of insulin-like growth factor binding proteins-1, -3, and -5 during early diabetic renal hypertrophy in rats. AB - The renal insulin-like growth factor-I (IGF-I) system has been implicated in the pathogenesis of renal hypertrophy, altered hemodynamics, and extracellular matrix expansion associated with early diabetes. The relative abundance of IGF binding proteins (IGFBPs) in the renal microenvironment may modulate IGF-I actions. However, the precise IGFBPs expressed in the glomerular and tubulointerstitial compartments during diabetic renal growth have not been characterized. In the present study, in situ hybridization studies were performed to examine the expression of IGFBP-1 to -6 messenger RNAs (mRNAs) 3, 7, and 14 days after streptozotocin (STZ) injection in rats. In control, nondiabetic kidneys, all six IGFBP mRNAs were differentially expressed with a predominance of IGFBP-5. The onset of renal hypertrophy in STZ-induced diabetes was associated with a rapid and site-specific induction of IGFBP-1, -3, and -5 mRNAs. In contrast, basal expression of IGFBP-2, -4, and -6 mRNAs was not altered in diabetic rats. IGFBP-5 mRNA expression increased in diabetic glomeruli, cortical, and inner medullary peritubular interstitial cells at days 3, 7, and 14. Although normal glomeruli failed to express IGFBP-3, it was induced concomitantly with IGFBP-5 in diabetic glomeruli and cortical peritubular interstitial cells. IGFBP-1 mRNA levels also increased in cortical tubular cells at each time point tested. Peak induction of IGFBP-3 and -5 was observed at day 3, whereas IGFBP-1 was delayed until day 7. IGFBP-1, -3, and -5 mRNA levels declined by day 14, but remained persistently elevated above control. By immunoperoxidase staining, similar alterations in the pattern of IGFBP-3 and -5 protein expression were observed at each time point. The preferential and site-specific increase in IGFBP-1, -3, and -5 suggest that these IGFBPs may regulate the local autocrine and/or paracrine actions of IGF-I and contribute to the pathogenesis of the early manifestations of diabetic nephropathy. PMID- 9856517 TI - Benefits of exercise training in patients on continuous ambulatory peritoneal dialysis. AB - We examined the effects of a 12-week exercise program on the exercise tolerance, blood biochemistry, blood pressure (BP) control, cardiac function, and quality-of life (QOL) scores in 13 patients undergoing continuous ambulatory peritoneal dialysis (CAPD; six men, seven women; mean age, 46.5+/-12.8 years; mean duration on dialysis, 4.8+/-3.8 years). The patients underwent exercise training on treadmill, bike, and arm ergometers thrice weekly. Seven CAPD patients matched for age, sex, and duration on dialysis served as controls. The mean peak aerobic capacity (VO2peak) of the exercisers increased by 16.2% after training (pre- and postexercise, 17.2+/-5.2 v 20.0+/-6.4 mL/kg/min; P=0.004). Although there were no significant changes in serum urea, creatinine, albumin, and hematocrit levels; left ventricular diastolic/systolic diameters; and ejection fraction, an increasing trend of high-density lipoproteins (HDLs) was observed in the exercisers (baseline v postexercise, 33+/-11 v 40+/-14 mg/dL; P=0.06). Twenty four-hour ambulatory BP monitoring showed a significant increase in daytime systolic BP in the exercisers (pre- and postexercise, 142+/-26 v 157+/-22 mm Hg; P=0.003), but no significant changes could be found in the ambulatory daytime diastolic BP, nocturnal BP, and resting clinic BP. The patients' QOL improved after training, with better scores in two Kidney Disease Quality of Life scales (KDQOL): burden of kidney disease and physical functioning. Two mild and uncomplicated hypotensive episodes were reported in two patients immediately after training. No changes occurred in exercise capacity, blood biochemistry, BP profile, and QOL scores in the controls. We conclude that structured aerobic exercise is safe and can improve the exercise tolerance and QOL outcomes in CAPD patients. PMID- 9856518 TI - Caloric intake in a group of peritoneal dialysis patients. AB - Dialysis patients as a group appear to have a caloric intake less than the recommended values; because their energy expenditure is not different than that of healthy adults, they should be in negative caloric balance and lose body mass progressively. We retrospectively analyzed our data in a group of peritoneal dialysis patients who had dietary evaluations and kinetic measurements performed two consecutive times over a period of 6 months. Body weight, lean body mass (LBM), fat mass, and anthropometric parameters remained stable over this period of time, suggesting that these patients are not in negative energy balance. When their daily caloric intake was normalized to a new ideal body weight derived from the LBM calculated from creatinine kinetics, the values were within normal limits, suggesting that these patients were in zero energy balance; hence, a stable body mass was expected. PMID- 9856519 TI - Cooling effect of continuous renal replacement therapy in critically ill patients. AB - Hypothermia is reported to increase intensive care unit (ICU) mortality. The heat loss that occurs during continuous renal replacement therapy (CRRT) favors the development of hypothermia. In an effort to assess the influence of CRRT on body temperature, we reviewed the records of 72 consecutive ICU patients treated with CRRT and further prospectively studied the temperature in the inlet and outlet lines for blood and dialysate of 27 other patients at various flow settings during continuous venovenous hemodialysis (CVVHD). Among the 72 retrospective cases, 36 episodes of hypothermia (core body temperature <35.5 degrees C) occurred and persisted for a mean of 2.6+/-1.8 days. It was more frequent during venovenous than arteriovenous modalities (31 of 67 v5 of 20, respectively); no patients developed hypothermia during arteriovenous slow continuous ultrafiltration (AVSCUS), whereas 48% of the patients undergoing CVVHD became hypothermic, occurring earlier in the therapy course (days 2 to 4). Mean arterial pressure (MAP) tended to increase after CRRT initiation, but absolute changes were not statistically significant. In the prospective arm, the CVVHD circuit temperatures were directly measured. Whereas no attempt was made to change body temperature, stepwise changes in blood (Qb) and dialysate flow rate (Qd) produced venous circuit temperature changes: the higher the Qb, the smaller the arteriovenous temperature differences independent of changes in Qd (P < 0.001). Also, venous circuit temperature varied directly with Qd at fixed Qb (P < 0.001). This relationship also held for temperature conversion to lost energy units per minute. Using room temperature dialysate, CRRT may significantly lower patients' core temperatures. Although the clinical significance of this effect is not clear at this point, energy loss during CVVHD may be important in hemodynamic stability or patient prognosis. PMID- 9856520 TI - High-flux dialysis lowers plasma leptin concentration in chronic dialysis patients. AB - Leptin is a protein produced by fat cells and involved in body weight regulation. Plasma leptin is significantly higher in some hemodialysis (HD) patients than in normal controls. We examined the influence of dialyzer membrane biocompatibility and flux on elevated plasma leptin concentrations in hemodialysis patients. Employing a crossover design, leptin and tumor necrosis factor-alpha (TNF-alpha) levels were serially determined in eight chronic dialysis patients. Patients were dialyzed sequentially on low-flux cellulosic (TAF) dialyzers, low-flux (F8) polysulfone, high-flux (F80B) polysulfone, then low-flux polysulfone and cellulosic dialyzers again. Mean leptin concentrations were similar when low-flux polysulfone or cellulosic dialyzers were employed (141.9+/-24.2 microg/L versus 137.8+/-18.4 microg/L, respectively (P=NS). In contrast, leptin fell significantly on the high-flux polysulfone dialyzer (99.4+/-16.2 microg/L) compared with cellulosic (P < 0.005), and low-flux polysulfone dialyzers (P < 0.02). Leptin clearance by the high-flux polysulfone dialyzer was significantly higher than the low-flux dialyzers (50.4+/-21.5 v -9.6+/-10.3 mL/min; P=0.043), but did not account fully for the 30% decline in plasma leptin during the high flux arm of the study. Concentrations of TNF-alpha were lower when high-flux polysulfone dialyzers were employed, but there was no correlation of individual TNF-alpha levels with leptin concentrations. High-flux dialysis lowers plasma leptin concentrations an average of 30%, but biocompatibility does not influence leptin levels. The decrease in plasma leptin on high-flux dialysis cannot be explained solely by enhanced clearance. PMID- 9856521 TI - Effect of intravenous alfacalcidol on lymphocyte phenotyping in hemodialysis patients. AB - Chronic renal failure is associated with decreased production of active vitamin D and also results in altered lymphocyte population. We studied the effect of alfacalcidol on lymphocyte phenotype. There were 15 patients (10 males, 5 females) with a mean age of 54.3+/-14.4 years who had been on chronic maintenance haemodialysis for a mean period of 3.2+/-1.5 years. Intravenous alfacalcidol was given three times weekly during dialysis for a duration of 6 months. Our results show a significant increase in NK cells from 0.20+/-0.12 to 0.27+/-0.28 (P=0.001), without a significant change in CD2, CD19, CD4, CD8 population, and CD4/CD8 ratio. PMID- 9856522 TI - Reinforced intradermal hepatitis B vaccination in hemodialysis patients is superior in antibody response to intramuscular or subcutaneous vaccination. AB - Since 1960, hepatitis B virus-associated chronic liver disease has been considered an important problem in dialysis units in both Europe and North America. Separate dialysis facilities for hepatitis B-infected patients, the implementation of universal precautions for the prevention of transmission, and the active immunization against hepatitis B have now reduced the yearly incidence to less than 0.05% in Western countries. However, only 50% to 60% of patients with renal insufficiency develop sufficient immune response after intramuscular hepatitis B vaccination. The aim of the current study was to determine whether the mode of vaccine application plays a role in vaccination response and whether increasing the vaccine dose of primary intradermal hepatitis B vaccination can reduce the number of vaccine injections in hemodialysis patients. We designed a prospective, randomized study of antibody responses to hepatitis B vaccine given intradermally, subcutaneously, or intramuscularly in 81 hemodialysis patients. Outcome measures were rates of seroconversion, mean levels of anti-Hbs antibodies, and antibody levels 8 years after vaccination. The results show that intradermal hepatitis B vaccination response with a higher vaccination dose than previously used in hemodialysis patients is superior to conventional intramuscular and subcutaneous vaccination and is also well tolerated. Five intradermal injections of 20 microg each induced the development of sufficient anti-Hbs antibody titer, which persisted in 70% of the patients over 3 years. PMID- 9856523 TI - All currently used measurements of recirculation in blood access by chemical methods are flawed due to intradialytic disequilibrium or recirculation at low flow. AB - Blood flows and recirculations with standard and reversed direction of lines were measured by chemical (urea and creatinine) and ultrasound dilution (saline) methods in 47 chronic hemodialysis patients. Thirty-seven patients had 47 dual lumen, central vein (CV) catheters: 32 were PermCath (Quinton Instruments Company, Seattle, WA), 6 were Access Cath (MEDCOMP, Harleysville, PA), 3 were Soft Cell PC (Vas Cath, Mississauga, Ontario, Canada) and 6 were SNIJ (experimental catheters). Three of these last catheters had the tip staggered 7 mm, and three had flush tips; PermCath, Access Cath, and Soft Cell PC catheters have the tips staggered 23 to 25 mm. Forty-six catheters were implanted into the superior vena cava/right atrium, and one catheter was implanted through the left saphenous vein into the left iliac vein. The catheters were studied 1 to 31 months after implantation (median, 3.0 months). Ten patients with arteriovenous (AV) graft access were also studied. The stop-flow method was used in catheter dialysis, and the slow-flow method was used to calculate recirculations in AV access dialysis with samples for systemic blood concentrations taken from arterial line both before and after samples from the arterial and venous lines. At 500 mL/min pump speed, actual blood flow was 436+/-18 mL/min (mean+/-SD; range, 407 to 464 mL/min) with standard direction of catheter lines. At 500 mL/min pump speed, the arterial chamber pressure was -330+/-48 mm Hg (mean+/-SD; range, -380 to -225 mm Hg, and the venous chamber pressure was 259+/-48 mm Hg (mean+/-SD; range, 140 to 310 mm Hg). Arterial chamber pressure was less negative, and venous chamber pressure was less positive with SNIJ catheters, which had larger internal diameter (2.1 mm) compared with the other catheters (2.0 mm). Recirculation varied with the catheter design and the location of the catheter tip. In the catheters with tip staggered more than 20 mm and with standard line connection at pump speeds of 50 mL/min and 500 mL/min, recirculations were approximately 1 % and 5%, respectively, when measured by the chemical method. In the same catheters with reversed lines, the recirculations were approximately 5% and 27%, respectively. Inflow failure catheters with reversed lines had similar recirculation values to those of well-functioning catheters with reversed lines. In catheters with tips staggered 7 mm, and with standard connection of lines, recirculations were approximately 3% and 8%, respectively, at pump speeds of 50 and 500 mL/min. With reversed lines, at the same pump speeds, the values were 7% and 12%, respectively. In flush-tip catheters, the recirculation was higher at a 50 mL/min pump speed (approximately 17%) than at a pump speed of 500 mL/min (approximately 13%). The ultrasound dilution method usually gave lower values than the chemical methods, most likely because of overestimation of recirculation by chemical methods. At least triplicate measurements are needed because single measurements by the ultrasound dilution method are associated with substantial variation. We conclude that both currently used methods (stop flow and slow flow) of taking systemic samples for measurements of recirculation by chemical methods are flawed because of disequilibrium and recirculation at low flow. PMID- 9856524 TI - Renal-retinal syndromes: association of retinal anomalies and recessive nephronophthisis in patients with homozygous deletion of the NPH1 locus. AB - Tapeto-retinal degeneration is frequent in patients with nephronophthisis. Association of the most severe forms of tapeto-retinal dystrophy with NPH identifies a syndrome described first by Senior et al and Loken et al. This syndrome is distinct on molecular grounds from pure renal nephronophthisis (NPH1), which has its gene locus mapped on chromosome 2q13. We describe three families with large homozygous deletion of the NPH1 locus in which mild to moderate ocular lesions due to tapeto-retinal degeneration coexisted and were correlated to renal defects. This new association of NPH1 with retinal dystrophy is characterized by focal lesions of retina and is pauci-symptomatic in clinical presentation. For this reason it may remain unrecognized in most NPH1 patients. PMID- 9856525 TI - Familial hemolytic-uremic syndrome in three generations. AB - The hemolytic uremic syndrome (HUS) and thrombotic thrombocytopenic purpura (TTP) are complex entities characterized by microangiopathic hemolytic anemia, thrombocytopenia, and variable impairment of renal function, occasionally complicated by neurological symptoms. In both syndromes, rare instances of familial forms have been reported. We present the case of a family in which signs and symptoms of HUS/TTP appeared in three generations. We also briefly review the literature on inherited forms of HUS/TTP and discuss the outcome of renal transplantation in adult patients with this syndrome. PMID- 9856527 TI - Successful treatment of acute inferior vena cava and unilateral renal vein thrombosis by local infusion of recombinant tissue plasminogen activator. AB - Renal vein thrombosis can occur as a complication of nephrotic syndrome. We present the case of a young man with nephrotic syndrome caused by minimal change disease who developed acute inferior vena cava and left renal vein thrombosis. He was treated initially with intravenous heparin. Because of the persistence of severe left flank pain and gross hematuria, local infusion of recombinant tissue plasminogen activator was tried, with resolution of thrombi and subsidence of symptoms. Functional preservation of the involved kidney is good, as indicated by Tc-99m DMSA scan (involved kidney, 47.4%; uninvolved kidney, 52.6%). Anticoagulation is usually recommended as the treatment of choice in renal vein thrombosis. We believe that in cases with critical presentations, such as bilateral involvement, extension into inferior vena cava, acute renal failure, pulmonary embolism or severe flank pain, thrombolytic therapy should be considered as a second-line treatment if good response is not obtained with heparin. PMID- 9856526 TI - Childhood membranous nephropathy, circulating antibodies to the 58-kD TIN antigen, and anti-tubular basement membrane nephritis: an 11-year follow-up. AB - Childhood membranous nephropathy (MNP) with anti-tubular basement membrane (anti TBM) nephritis is a rare disorder that may have extrarenal manifestations. This article describes a new case to be added to the 10 previously reported. A renal biopsy specimen from a 1-year-old white boy with nephrotic syndrome, microhematuria, and hypertension showed MNP (granular global IgG, IgA and C3, and segmental IgM and C1q) associated with hypercellularity and granular deposits of IgM and C1q in the mesangium, arteriolar IgA, and linear TBM IgG, IgA, and C3. A biopsy at age 4 years showed MNP (IgG and C3) and linear IgG and C3 along the TBM. Six months later, temporary glucosuria suggested a mild tubular dysfunction. Biopsy at age 8 years showed sclerosing MNP (IgG and C3), linear TBM IgG and C3, and chronic active tubulointerstitial nephritis (TIN). Indirect immunofluorescence showed circulating anti-TBM antibodies, and the enzyme-linked immunosorbent assay (ELISA) approach verified strong reactivity with the 58-kd TIN antigen. Despite trials with steroids, chlorambucil, azathioprine, and cyclosporine, end-stage renal disease developed by the age of 9 years. At age 10 years, the patient received a cadaveric kidney transplant. With the patient now aged 12 years, the graft is still functioning well, without any clinical evidence of disease recurrence. Neurological, ocular, and abdominal symptoms, including nonbacterial diarrhea, were observed during the follow-up period. The pathophysiology of these extrarenal symptoms remains unclear. Serotyping and genotyping of HLA antigens (A2, A10, B12, B41, DR5 [1101, 1103-4, 1106 or 1108 1113], DR6 [1303, 1312, or 1413], DRB3 [*0101 and 0201-2 or 0301], DQA1 [*0501 homozygous], and DQB1 [*0301 homozygous]) did not indicate any HLA association similar to those described previously in childhood MNP with anti-TBM nephritis (HLA-B7 in four patients, HLA-DR8 in two patients). The presented case is the fifth in the literature that displays reactivity with the 58-kd TIN antigen, and for which data on HLA antigens are reported. PMID- 9856529 TI - Molecular biology and the kidney. PMID- 9856528 TI - PKD and CF: an interesting family provides insight into the molecular pathophysiology of polycystic kidney disease. PMID- 9856530 TI - Intravenous infusion of total dose iron in peritoneal dialysis patients: a critical appraisal. PMID- 9856531 TI - H+-ATPase in hyperkalemic distal renal tubular acidosis. PMID- 9856532 TI - A 46-year-old woman with nephrotic syndrome and persistent microscopic hematuria. PMID- 9856534 TI - Physiology of beer or non-beer potomania. PMID- 9856533 TI - Uteroglobin: physiological role in normal glomerular function uncovered by targeted disruption of the uteroglobin gene in mice. AB - Blastokinin or uteroglobin (UG) is an evolutionarilly conserved, steroid inducible, homodimeric, multifunctional, secreted protein with potent Immunomodulatory/antiinflammatory properties. Recently, a UG-receptor expressed on several malignant and normal cell types has been characterized. Although the biochemistry, structural, and molecular biology of UG have been extensively studied, its physiological function(s), until recently, remained unknown. By generating UG-null (UG-/-) mice, we determined that an essential role of UG is to prevent severe renal disease caused by an abnormal deposition of predominantly multimeric fibronectin (Fn) and collagen in the glomerulus. The molecular mechanisms by which UG prevents this disease in control (UG+/+) mice, at least in part, is attributable to its high-affinity binding to Fn and the formation of Fn UG heteromers, which counteract both Fn-Fn and Fn-collagen interactions, required for abnormal tissue deposition. In addition, by inhibiting secretory phospholipase A2 (sPLA2) activity and decreasing the level of lysophosphatidic acid (LPA), UG may indirectly prevent the activation of integrins (eg, alpha5beta1) that enhance abnormal tissue deposition of Fn. The mechanism(s) of UG action is likely to be even more complex, because it also functions through a receptor-mediated pathway that has not yet been clearly defined. Nevertheless, the UG gene-knockout mice provide a valuable animal model for investigation of human glomerulopathies in general and familial Fn-deposit glomerulopathy in particular. PMID- 9856535 TI - Epidemiology of work-related aviation fatalities in Alaska, 1990-94. AB - Alaska, with less than one-half of 1% of the United States workforce, accounts for 9% of all occupational aviation fatalities nationally; 30% of all occupational fatalities in Alaska are related to aviation. To understand this high mortality, we investigated occupational aviation crashes to identify risk factors. Occupational aviation fatalities in Alaska during 1990-94 were examined using National Transportation Safety Board reports and merged with records from the Alaska Occupational Injury Surveillance System. There were 876 aircraft crashes; 407 (46%) were work-related. Occupational crashes were 2.2 times (CI: 1.5, 3.2) more likely to result in fatalities than non-occupational crashes. Risk factors identified included poor weather conditions defined as Instrument Meteorological Conditions (IMC). A crash during IMC was 5.3 times (CI: 3.5, 7.9) more likely to result in fatalities than crashes in other conditions. Of aircraft involved in fatal occupational incidents, 33% were not completely destroyed, allowing the potential for survivors. An estimated 30% reduction in fatalities could have occurred if current technology in occupant protection had been used. PMID- 9856536 TI - Long term disability rates in a cohort of Air Canada pilots. AB - Causes of long term disability (LTD) were collected between January 1, 1981 and December 31, 1990 among all active pilots in a cohort of Air Canada pilots. During this 10-yr period, 474 (20.9%) of the 2271 pilots had LTD; there were 1107 cases of LTD corresponding to a total duration of leave of absence of 4161 mo. Frequencies of the main diagnostic categories were: injuries: 249 (22.5%); mental disorders: 175 (15.8%); circulatory system: 157 (14.2%); digestive system: 128 (11.6%); and musculoskeletal system: 121 (10.9%). LTD rates increased with age, ranging from 1.86-9.22 per 1000 pilots per year from the younger (20-29 yr) to the older age group (50-59 yr), respectively. Several causes of LTD could be potentially preventable, in particular injuries which accounted for 63.6% of all causes in the younger age group and remained a significant condition at subsequent ages. PMID- 9856537 TI - Male/female SACM endurance comparison: support for the Armstrong Laboratory modifications to the CSU-13B/P anti-G suit. AB - BACKGROUND: The standard anti-G suit (CSU-13B/P) was designed based on male body structure. Females differ from males with respect to body proportionality. In Armstrong Laboratory (AL) studies, females have terminated centrifuge simulated air combat maneuvers (SACM) because of anti-G suit (CSU-13B/P modified according to original T.O. 14P3-6-121)(OTO) discomfort. AL modifications to the suit have since been adopted in the OTO in an attempt to provide females a best-fit suit (AL Mod). The study examined male/female SACM endurance with females wearing both the OTO and the AL Mod suits. METHODS: There were 6 females and 8 males who performed a +5.0 to +9.0 Gz SACM to fatigue using the anti-G straining maneuver with anti-G suit inflation. The females performed in both the OTO and AL Mod suits while the males performed in the OTO suit only (OTO was their best-fit suit). RESULTS: Wearing the OTO, males performed the SACM significantly longer than the females, three of whom reported severe suit discomfort. However, when the females wore the AL Mod suit, their SACM endurance almost doubled over their OTO performance and none reported suit discomfort. When wearing their best-fit suits, there was no significant gender difference in SACM endurance. CONCLUSIONS: These data support the efficacy of the AL modifications to the CSU-13B/P anti-G suit through greatly improved performance during the +5.0 to +9.0 SACM in females. These data also suggest that, in the small sample examined, when fitted with a best-fit anti-G suit, females can endure the +5.0 to +9.0 SACM to the same degree as males. PMID- 9856538 TI - Using functional magnetic resonance imaging to understand the mechanisms of consciousness. AB - BACKGROUND: In order to elucidate mental functions that subserve human consciousness, brain activation was investigated in 12 normal, right-handed volunteers who performed tasks of selective attention, working memory, and sensorimotor coordination during the collection of multislice echoplanar functional magnetic resonance images. HYPOTHESIS: These functions are located in (and controlled by) distinct anatomical regions that can be identified by functional magnetic resonance imaging techniques. METHODS: In each subject, 100 10-slice data sets were acquired using a 1.5-T scanner and the blood oxygenation level dependent contrast technique. Time-series regression modeling estimated power in the magnetic resonance signal during the on/off phases of task performance. Comparison between subjects was made possible by the transformation of each data set into standard Talairach space. RESULTS: Activation maps were based on the median value of the fundamental power quotient at each voxel. Results showed the activation of prefrontal and parasagittal cortices in both the selective attention and working memory tasks, but they also revealed activation in both insular cortices and the posterior cingulate gyri. CONCLUSIONS: The results provide evidence for structures in the anterior right hemisphere and left medial frontal lobe for attentional tasks, although there appears to be an engagement of a widespread network of anterior brain structures, possibly with the inhibition of some posterior regions, during task performance. The sensorimotor coordination task showed activation regions similar to those seen in selective attention. Once learned, this task probably demands attention rather than overt conscious motor control. Clearly, the functions of attention, working memory, and sensorimotor coordination are not located in single, discrete brain areas. However, interactions and interplay between related areas were demonstrated, giving supporting evidence that complex mental operations rely on the coordinated activity of widely distributed brain regions that contribute to neural networks. PMID- 9856540 TI - Optokinetic nystagmus correlates with severity of vection-induced motion sickness and gastric tachyarrhythmia. AB - PURPOSE: The present study investigated the relationship of the frequency of nystagmus to the severity of optokinetic rotation-induced motion sickness. METHODS AND RESULTS: Eighty-seven subjects viewed a vertically striped, rotating drum for 16 min. Subjects' electrooculograms, ratings of vection, and subjective symptoms of motion sickness (SSMS) were measured during the drum rotation period. Subjects' electrogastrograms (EGGs) were recorded throughout the baseline and drum rotation periods. The ratios of the spectral intensity of EGG 4-9 cycles per minute (cpm) between drum rotation and baseline periods were calculated. The results indicated that the frequency of nystagmus was positively correlated with the ratings of vection (r = 0.40, p < 0.0001), scores of SSMS (r = 0.61, p < 0.0001), and ratios of EGG 4-9 cpm spectral intensity between drum rotation and baseline periods (r = 0.54, p < 0.0001). CONCLUSION: These results demonstrated that more rapid eye movement indexed by higher frequency of optokinetic nystagmus are related to the development of symptoms of motion sickness and gastric tachyarrhythmia. PMID- 9856539 TI - Drinking milk or water has no effect on the severity of optokinetic rotation induced symptoms of motion sickness. AB - BACKGROUND: The present study investigated the effects of drinking milk or water before viewing an optokinetic rotation drum on the severity of motion sickness and abnormal gastric myoelectric activity. METHODS: There were 27 subjects who drank 300 ml of milk or water, or drank nothing, in three separate sessions with 2-d intervals between every two sessions. Subjects in each session then viewed a vertically striped optokinetic rotation drum for 16 min. Symptoms of motion sickness and electrogastrograms (EGGs) were measured. RESULTS: Subjects reported no significant difference in symptoms of motion sickness among sessions of drinking milk, water, or nothing. EGG recordings indicated that subjects in sessions of drinking milk and water significantly increased normal gastric myoelectric activity at EGG 3 cycles per minute (cpm). Subjects in all three sessions increased abnormal EGG activity at 4-9 cpm during drum rotation period, however, the ratios of EGG 4-9 cpm of drum rotation and baseline periods were not significantly different among sessions. CONCLUSION: These results demonstrated that drinking milk or water before viewing an optokinetic rotation drum exhibited no effect on the severity of symptoms of motion sickness and abnormal gastric myoelectric activity. PMID- 9856541 TI - The optokinetic cervico reflex during simulated helicopter flight. AB - INTRODUCTION: The optokinetic cervico reflex (OKCR) is a recently hypothesized, visually driven reflex that serves to stabilize the image of the external horizon on the retina during roll maneuvers in high-performance aircraft. Although reported anecdotally, head tilt during helicopter flight has not been studied formally. Such research is required to determine the full impact and significance that it may have on the flying performance of a rotary-wing aviator. OBJECTIVE: The aim of this study was to investigate the relationship between horizon position and the perception of orientation and, thus, to generate vital information to assess whether OKCR plays an important role in spatial disorientation. HYPOTHESIS: Pilots of rotary-wing aircraft will exhibit the OKCR. METHODS: A UH-60 flight simulator study, with 20 volunteer pilots participating, was performed to examine the effects of this reflex during day flight and during flight with night vision goggles (NVGs). RESULTS: The results confirm that the OKCR occurs during simulated helicopter flight, both with and without NVGs. As with previous studies, head roll increased during flight under visual meteorological conditions in relation to an increasing aircraft roll angle up to a maximum sustainable level and then remained constant. Head roll did not occur during flight under instrument meteorological conditions. CONCLUSION: The presence of the OKCR will impact rotary-wing operations. Various aspects are discussed, and recommendations are made for future research. PMID- 9856542 TI - Flight coverall microclimate evaluation using a Japanese type sweating mannequin. AB - BACKGROUND: It is important to examine the microclimate of a pilot's clothing to evaluate the heat stress the clothing imparts on a pilot, but problems arise with regard to individual variation (phenotype of the pilot, sweating dose, wear condition of the flight coveralls, etc.). HYPOTHESIS: The present study used a Japanese type sweating mannequin (TOM-III) to evaluate the microclimate of flight coveralls. METHODS: TOM-III (TOYOBO, Co., Ltd., Tokyo, Japan) has a characteristic Japanese human body type, and the sweat rate and body temperature can be regulated (240 g of sweat per hour, 36.0 degrees C). The microclimate of the clothing was evaluated by means of 20 temperature/humidity sensors located on the surface of the mannequin skin. TOM-III wore four different assemblies of clothing (CWU-66/P USAF chemical defense flight coverall; JASDF summer-type flight coverall which is similar to the nonchemical defense USAF flight coverall; ordinary 100% cotton underwear; and CWU-66/ P with cotton underwear). We put TOM III in climatic chamber (21.2 +/- 0.5 degrees C, 50.0 +/- 3.0% relative humidity (RH), 0.1 m air flow x s(-1), dressed in test clothing, and measured the inside temperature (IT) and inside relative humidity (IRH) of the clothing for 60 min (10 min non-sweating, 30 min sweating and 20 min non-sweating). RESULTS: Approximately 5 min after the onset of sweating, the IRH of the CWU-66/P (38.0%) was lower than JASDF flight coverall (42.1%; p < 0.01). At the end of the sweating period, the IRH of the CWU-66/P and the JASDF flight coverall were 46.2% and 52.6%, respectively (p < 0.01). The results indicated that the CWU-66/P flight coverall was better suited for heat stress than the JASDF summer-type flight coverall. CONCLUSIONS: TOM-III may be useful for microclimate evaluation and/or the development of clothing without considering individual variation against various climatic conditions. PMID- 9856543 TI - Thermoregulatory effects of caffeine ingestion during submaximal exercise in men. AB - BACKGROUND: The exclusive effect of caffeine ingestion on exercise thermoregulation is unclear; data indicate that caffeine may have a positive effect, a negative effect, or no effect. METHODS: Rectal (TRE) and mean skin (TSK) temperatures, skin heat conductance (HSK), and sweat rate (MSW) were measured during 30 min of rest and subsequent 70 min of submaximal cycle ergometer exercise (67% VO2PEAK) in 11 aerobically conditioned men (mean +/- SD 29 +/- 6 yr, 49 +/- 6 mL x min(-1) x kg(-1) VO2PEAK) under two conditions: a caffeine (10 mg x kg(-1) ingestion (CI) session and a noncaffeine ingestion (NCI) control session. RESULTS: There were no significant differences in physiological or thermoregulatory parameters during exercise: X (+/-SE) end exercise levels for the NCI and CI sessions, respectively, were VO2 = 2.50 +/- 0.09 vs. 2.55 +/- 0.09 L x min(-1); heart rate = 145 +/- 7 vs. 145 +/- 5 bpm; HSK = 30 +/- 3 vs. 28 +/- 3 kcal x m(-2) x h(-1) x degrees C(-1); MSW = 393 +/- 35 vs. 378 +/- 36 g x m(-2) x h(-1); and TRE = 38.3 +/- 0.2 vs. 38.4 +/- 0.1 degrees C. Control TSK was lower than that for CI by 0.4 to 0.5 degrees C at rest and during exercise. CONCLUSION: Ingestion of a high level (10 mg x kg(-1) of caffeine has no effect on skin heat conductance, sweating, or the rate of increase and final level of rectal temperature during moderate, submaximal leg exercise. PMID- 9856544 TI - Arterial oxygen saturation for prediction of acute mountain sickness. AB - BACKGROUND: Acute mountain sickness (AMS) is a usually self-limiting syndrome encompassing headache, nausea and dizziness. AMS is seen in those that go from low to high altitudes too quickly, without allowing sufficient time to acclimatize. At present, susceptibility to AMS cannot be predicted. One feature of AMS noted in some studies is impaired gas exchange. If impaired gas exchange presages AMS then those individuals with exaggerated hypoxemia at high altitude may be more likely to develop AMS. If true, then monitoring of arterial oxygen saturation (SaO2%) may differentiate AMS-resistant individuals from those with impending AMS. METHODS: To test this hypothesis, we measured SaO2% and AMS symptom scores in 102 healthy asymptomatic climbers at 4200 m on Denali (Mt. McKinley) prior to their further ascent toward the summit at 6194 m, and on their return from higher altitudes to 4200 m. RESULTS: The results show that exaggerated hypoxemia in asymptomatic climbers prior to further ascent correlates with subsequent AMS (r = -0.48, p < 0.001). Criteria are presented for identification of 80-100% of those climbers who later become ill with AMS. CONCLUSION: We conclude that resting arterial hypoxemia is related to later development of clinical AMS, and can exclude the occurrence and caution those at risk for development of subsequent AMS. Likely mechanisms are hypoventilation relative to normally acclimatizing individuals and/or abnormalities of gas exchange. Thus, non-invasive oximetry provides a simple, specific indicator of inadequate acclimatization to high altitudes and impending AMS. PMID- 9856545 TI - Assessment of acute mountain sickness by different score protocols in the Swiss Alps. AB - INTRODUCTION: Purpose of the present study was to evaluate the Lake Louise acute mountain sickness (AMS) score questionnaire at different altitudes and to compare it with the currently used clinical score and the environmental symptoms questionnaire AMS-C score. METHODS: We investigated 490 climbers who stayed over night at 4 huts in the Swiss Alps, located at the altitudes of 2850 m, 3050 m, 3650 m, and 4559 m. AMS was assessed using our previously described clinical score, the Lake Louise consensus AMS score questionnaire and the environmental symptoms questionnaire III. RESULTS: Below 4000 m, the prevalence of AMS, defined by symptoms that force a reduction in activity, was 7%; when assessed with the clinical score (score > or = 3) it was 22%; with the AMS-C score (score > or = 0.7) 4% and with the Lake Louise score (score > 4) 8%. At the altitude of 4559 m, the prevalence of AMS was 30%, 38%, 40%, and 39%, respectively. The standardized regression coefficients from multiple regression analysis (adjusted R2 0.65, p < 0.001) were 0.45 (p < 0.001) for the self-reported Lake Louise score, 0.48 (p < 0.001) for the sum of the points assigned in the clinical section of the Lake Louise questionnaire, and 0.05 (p = 0.27) for the AMS-C score. The sensitivity and specificity of the Lake Louise score > 4 was 78% and 93%, respectively. CONCLUSIONS: The Lake Louise consensus score is adequate and, compared with the AMS-C score, more effective for the assessment of acute altitude illness at different altitudes. PMID- 9856546 TI - Prevention of minor neck injuries in F-16 pilots. AB - INTRODUCTION: F-16 pilots have a high incidence of minor neck injuries. It was hypothesized that pilots who did neck strengthening exercises and pilots who used other preventive strategies would have fewer injuries. METHOD: We surveyed 268 U.S. Air Force F-16 pilots. Subjects were divided into two groups. Group I, the Early Intervention Group, performed an intervention, or not, from the start of their F-16 careers. Outcomes were measured as a percent of pilots reporting an injury during their F-16 careers. Group II, the Midstream Intervention Group, initiated an intervention after sustaining an injury. Injuries before and after the intervention were compared as a median injury rate per 100 h F-16 time. RESULTS: The 1 -yr prevalence of neck injury was 56.6% and for an F-16 career was 85.4%. For every 100 h in the F-16, the risk of injury increased by 6.9%. Only 26.9% of the pilots routinely did neck strengthening exercises. For the Early Intervention Group, fewer injuries were associated with neck strengthening exercises and placing the head against the seat prior to loading +Gz. For the Midstream Intervention Group, a lower median injury rate was associated with neck strengthening exercises, placing the head against the seat prior to loading, warming up with stretching or isometrics, prepositioning the head prior to loading, and unloading prior to moving the head. Interventions not associated with fewer injuries included body exercises and placing the head against the canopy. CONCLUSION: Certain strategies may prevent neck injuries. Prospective research is needed to confirm these results. PMID- 9856547 TI - Design and evaluation of a portable rigid forced-air warming cover for prehospital transport of cold patients. AB - Forced-air warming is promising for use during transport of cold patients. However, the current soft covers may be easily damaged in the nonhospital environment. Two rigid covers were designed to direct heat to the torso and thighs. Five subjects (one female) were heated with an AC powered heater (Bair Hugger 505, Augustine) and either a soft cover or the rigid covers (with heat input at the head or abdomen). Compared to the soft cover, the rigid cover (with heat input at the abdomen) provided similar heat delivery but a higher mean skin temperature. We conclude that the portable rigid covers are efficient for treatment of cold victims during pre-hospital transport. PMID- 9856548 TI - Operation Homecoming: 25 years later. AB - Following an end to U.S. involvement in the Vietnam War in early 1973, approximately 600 Army, Navy, Air Force, and Marine personnel, the great majority aviators, were released from captivity in North and South Vietnam and Laos. Their initial medical evaluation was performed at USAF Hospital, Clark Air Base, Republic of the Philippines, between February and April 1973. The author describes the events of those memorable days. PMID- 9856549 TI - The return of Vietnam POWs--the aeromedical phase of Operation Homecoming 1973. PMID- 9856550 TI - Psychiatric issues affecting long duration space missions. AB - BACKGROUND: As we move into the next Millennium, increasing numbers of people will travel into space. Psychological screening methods will be relaxed on near Earth missions, such as might occur on a space station or a lunar colony. Crewmembers on interplanetary missions such as a trip to Mars will have to deal with psychiatric problems themselves with no possibility of evacuating an affected individual. For these reasons, it is important for support personnel on Earth and crewmembers in space to be knowledgeable about psychiatric difficulties that might occur and their appropriate treatments. METHODS: Anecdotal reports from long duration manned space missions and studies from space analog environments on Earth were reviewed for information concerning these issues. RESULTS: Psychiatric problems pertinent to the space environment include adjustment and psychosomatic reactions, asthenia, mood and thought disorders, and post-mission personality changes and family problems. Counter-measures to ameliorate these difficulties involve pre-launch selection and training; monitoring, support, and counseling/psychotherapy during the mission; and post return debriefings with crew members and their families. Psychoactive medications have been useful during space missions, although unclear pharmacokinetic issues related to microgravity need to be taken into account. CONCLUSION: Although much is known about psychiatric issues related to long duration manned space travel, more empirical work needs to be done during actual space missions. PMID- 9856551 TI - You're the flight surgeon. 35-year-old helicopter pilot with ehrlichiosis. PMID- 9856552 TI - You're the flight surgeon. C-130 pilot with supraspinalus muscle tear. PMID- 9856553 TI - Aerospace medicine and ethics. PMID- 9856554 TI - Long first metacarpal in monozygotic twins with probable Baller-Gerold syndrome. AB - We report on a pair of monozygotic twins with probable Baller-Gerold syndrome (BGS). Twin A had severe coronal craniosynostosis. Twin B had right radioulnar and ipsilateral first metacarpal hypoplasia. Both had bilateral fifth finger clinodactyly. Assuming that the twins were truly monozygotic, a single genetic disorder (i.e., BGS) could explain the variable expression. Together the twins have the typical anomalies of BGS. The diagnosis was supported by the metacarpophalangeal profile (MPP) which confirmed hypoplasia of the first right metacarpal in Twin A and bilateral fifth finger brachymesophalangy in both twins. Furthermore, the MPP showed an unexpected abnormal lengthening of the first metacarpal (unilateral in Twin A and bilateral in Twin B), a previously undetected radial ray defect in BGS. These findings suggest the possibility that the MPP may assist recognition of mild cases of BGS such as those with apparently isolated craniosynostosis or isolated upper limbs defects. PMID- 9856555 TI - Blepharophimosis, hypoplastic radius, hypoplastic left heart, telecanthus, hydronephrosis, fused metacarpals, and "prehensile" halluces: a new syndrome? AB - We report on the prenatal ultrasound and postnatal findings in an infant born to a healthy, nonconsanguineous couple. The infant had microcephaly, telecanthus, blepharophimosis, cleft palate, micrognathia, abnormally modeled ears, hypoplastic left heart, hypoplastic radii and ulnae with radial subluxation, pseudoarthrotic distal humeri, fused metacarpals, tibial bowing, unusual feet with long halluces, hydronephrosis, patent urachus, abnormal electroencephalogram, and normal karyotype. To our knowledge, this combination of anomalies has not been recognized previously and may represent a new condition. PMID- 9856556 TI - Different patterns of obstetric complications in myotonic dystrophy in relation to the disease status of the fetus. AB - The obstetric histories of 26 women with myotonic dystrophy (DM), who had a total of 67 gestations, were reviewed retrospectively comparing gestations with affected (DM-fetuses) and unaffected fetuses (UA-fetuses). Second, the influence of gestation on the disease course and the personal attitude towards family planning in DM was assessed. Miscarriages and terminations occurred in 11 pregnancies. Of the 56 infants carried to term, 29 had or most likely had inherited the gene for DM from their affected mothers at the time of investigation; 18 (61%) in this series were affected by the congenital form of DM. Perinatal loss rate was 11% and associated with congenital DM. The rate of obstetric complications was significantly increased in all women. However, preterm labor was a major problem in gestations with DM-fetuses (55 vs. 20%), as was polyhydramnios (21% vs. none). While forceps deliveries or vacuum extractions were required in 21% of deliveries with DM-fetuses and only 5% of UA-fetuses, the frequency of Cesarean sections was similar in both groups (24 and 25%). Obstetric problems were inversely correlated with age at onset of maternal DM, while no effect of age at delivery or birth order on gestational outcome was seen. DNA analysis confirmed the diagnosis in 19 patients by the presence of enlarged CTG repeats (EcoRI-expansions) on chromosome 19. Of the 17 patients whose CTG repeat length was known, 59% were classified as E2 (corresponding to 500-1000 repeats), 24% as E1 (<500 repeats), while larger expansions (E3; 1000-1500 repeats, or E4; >1500 repeats) were seen in three patients (17%). Obstetric complications or congenitally affected children occurred in all maternal phenotypes and CTG repeat classes. Eight (31%) patients experienced a worsening of symptoms that was temporary, weight related in three cases, and persistent in five. With the exception of three patients, most new mothers were able to care for their families. To conclude, pregnant women with DM need constant obstetric monitoring and should be advised to deliver in centres with perinatal facilities. PMID- 9856557 TI - Atypical case of Smith-Lemli-Opitz syndrome: implications for diagnosis. AB - Smith-Lemli-Opitz (SLO) syndrome is an autosomal recessive disorder comprised of recognizable facial abnormalities, growth retardation, and multiple congenital anomalies, commonly involving genitalia, second and third toe syndactyly, and cleft palate. The condition is associated with hypocholesterolemia and elevated levels of 7-dehydrocholesterol (7DHC) resulting from deficient activity of the enzyme 7-dehydrocholesterol reductase. The clinical spectrum of SLO ranges from individuals with mental retardation and minor anomalies to those with major structural defects and early or even prenatal lethality. Low maternal serum unconjugated estriol (uE3) levels and a variety of fetal ultrasound anomalies have been identified in affected pregnancies, and prenatal diagnosis is possible by measurement of amniotic fluid 7DHC levels in pregnancies known to be at risk because of a previously affected child. We report on a pregnancy with low maternal uE3 level, abnormal antenatal ultrasound findings including limb deformities, ventriculomegaly, and hydrops fetalis, and a normal 46,XY karyotype. The infant died at birth. At autopsy the infant had hydrops, unusual face, cleft palate, genital abnormalities, Dandy-Walker malformation, and absence of toe syndactyly. Tests performed on cultured skin fibroblasts showed elevated levels of 7DHC and abnormalities of cholesterol biosynthesis characteristic of the metabolic defect that causes SLO. The atypical findings of hydrops, uncharacteristic facial appearance, and absence of toe syndactyly in this case additionally illustrates the wide phenotypic spectrum of SLO and the need for a high index of suspicion for a disorder with great clinical variability. Identification of another affected pregnancy with a low maternal uE3 level and abnormal fetal ultrasound findings in the presence of a normal karyotype lends additional support for consideration of prenatal biochemical testing for SLO in pregnancies with these findings, including pregnancies not previously known to be at risk. PMID- 9856558 TI - Novel mutations of the glutaryl-CoA dehydrogenase gene in two Japanese patients with glutaric aciduria type I. AB - We identified three different point mutations in the glutaryl-CoA dehydrogenase (GCDH) gene in two unrelated Japanese patients with glutaric aciduria type I (GA I). One patient was a homozygote for Arg355His and the other a compound heterozygote for Ser305Leu and Met339Val. Arg355His and Met339Val are mutations hitherto undescribed, and all three mutations are predicted to alter the secondary structure of GCDH. Molecular analysis is useful for definite diagnosis and/or prenatal diagnosis of GA-I. PMID- 9856559 TI - Low rates of pregnancy termination for prenatally diagnosed Klinefelter syndrome and other sex chromosome polysomies. AB - Over the past 9 years we counseled 55 couples whose unborn child was found to carry a sex chromosome polysomy. We performed a survey of postcounseling parental decisions about continuation or termination of these pregnancies. Of the 55 embryos or fetuses, 23 had the karyotype 47,XXY, 10 had 47,XYY, and 12 had 47,XXX. In addition, there were 10 instances of true mosaicism, i.e. 47,XXY/46,XY (n = 5), 47,XYY/46,XY (n = 2), or 47,XXX/46,XX (n = 3). Mean gestational age (+/ standard deviation) at diagnosis was 18.3+/-3.0 weeks. After comprehensive genetic counseling 48 (87.3%) of these pregnancies were carried to term. In seven cases (12.7%) the parents elected a pregnancy termination. Two of 31 pregnancies (6.5%) primarily ascertained at our center were aborted, whereas amongst the 24 referred cases, 5 couples (20.8%) opted for a termination. The mean gestational age of the terminated pregnancies was 19.7 weeks. The overall termination rate of 12.7% appears low in comparison with literature data. Most reports from other institutions present termination rates between 32 and 66%. The reason for the low rate of induced abortions in our study cohort is not clear. Cultural differences in parental perception of sex chromosomal polysomies may be of importance, and peculiarities of genetic counseling at our institution could also play a role. Although counseling was nondirective, we did put emphasis on providing prospective parents with information from unbiased follow-up studies of children with Klinefelter syndrome and other sex chromosome polysomies. PMID- 9856560 TI - Renal tubular dysgenesis, absent nipples, and multiple malformations in three brothers: a new, lethal syndrome. AB - We report on three brothers with renal tubular dysgenesis and absent nipples, each also had other malformations including pre-auricular pits and a preauricular tag, branchial clefts, choanal atresia, pulmonary lobation anomaly, ventricular septal defect, type IIB interrupted aortic arch, absent gallbladder, absent thymus, parathyroid gland, accessory spleen, imperforate anus, clinodactyly, and broad digits and small nails. All three infants died neonatally. This pattern of clinical malformations appears to be a previously unreported syndrome. PMID- 9856561 TI - Mutation analysis of Gaucher disease patients from Argentina: high prevalence of the RecNciI mutation. AB - Gaucher disease (GD) is caused by a deficiency of beta-glucocerebrosidase activity mainly due to mutations in the gene coding for the enzyme. More than 100 mutations have been identified to date and their frequencies have been established in several populations, including Ashkenazi Jews, among whom the disease is particularly prevalent. In order to study the molecular pathology of the disease in patients from Argentina, we conducted a systematic search for mutations in the glucocerebrosidase gene. Genomic DNA from 31 unrelated GD patients was screened for seven previously described mutations: N370S (1226A- >G), L444P (1448T-->C), D409H (1342G-->C), R463C (1504C-->T), 1263de155, RecNciI, and RecTL. This allowed the identification of 77.4% of the GD alleles: N370S and RecNciI were the most prevalent mutations found (46.8% and 21% respectively). Southern analysis demonstrated three distinct patterns for the RecNciI alleles. In order to identify the remaining alleles, the full coding region of the gene, all the splice sites, and part of the promoter region were analyzed by single strand conformational polymorphism analysis (SSCP) after polymerase chain reaction amplification. This extensive screening allowed the identification of 13 different mutations, accounting for 93% of the total number of GD alleles. Three novel missense mutations, I161S (599T-->G), G265D (911G-->A), and F411I (1348T- >A), were detected. Twelve polymorphic sites within the glucocerebrosidase gene are in complete linkage disequilibrium and define two major haplotypes, "-" and "+". Mutation N370S was always associated with the "-" haplotype, as described in other populations. Interestingly, the RecNciI alleles with the same Southern-blot pattern were always associated with the same haplotype. PMID- 9856562 TI - Four novel mutations of the connexin 32 gene in four Japanese families with Charcot-Marie-Tooth disease type 1. AB - DNA-based mutation analysis on the connexin 32 gene was performed in 49 families with Charcot-Marie-Tooth disease (CMT) type 1 but without duplication involving the chromosomal region, 17p12-p11.2. Mutations were identified in five of the 49 families, and four of the five mutations were hitherto undescribed: Va137Met, Glu57His, Arg142Glu, Val177Ala. X-linked CMT sometimes lacks evidence for X linked transmission and cannot be differentiated from CMT type 2, especially in females with mildly decreased nerve conduction velocity. Therefore, molecular analysis is useful for molecular pathology of their disease. PMID- 9856563 TI - Absence of correlation between skewed X inactivation in blood and serum creatine kinase levels in Duchenne/Becker female carriers. AB - The pattern of X inactivation in lymphocyte DNA was investigated in 107 Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) carriers (102 asymptomatic and 5 manifesting carriers) and 117 normal female controls of different ages, with the aim: a) to analyze the pattern of X inactivation in blood DNA of a large number of DMD/BMD carriers as compared to normal female controls; b) to determine if there is a decrease in serum creatine kinase (CK) levels with age in obligate DMD/BMD carriers; c) to determine if there is a correlation between X-chromosome inactivation and serum CK among asymptomatic DMD/BMD carriers of different ages or with different clinical manifestations in symptomatic carriers. A high proportion of females showed extremely skewed X inactivation (>90% of one X preferentially inactivated), which was almost the same among carriers and normal controls (19 and 24%, respectively). The mean serum CK was significantly greater among young (<20 years old) than adult (>20 years old) DMD/BMD carriers and it decreased significantly until age 20 with an apparent stabilization afterwards. No statistically significant correlation was found between the proportion of active X(DMD) in blood and serum CK activity in DMD/BMD carriers although it was higher among those less than 20 years old. Our observations suggest that highly skewed X-chromosome pattern in blood (with preferential inactivation of the X(N) chromosome) is not enough to predict that a young DMD carrier will develop muscular weakness. PMID- 9856564 TI - Severe congenital anomalies requiring transplantation in children with Kabuki syndrome. AB - Kabuki syndrome (KS) is a rare multiple malformation disorder characterized by developmental delay, distinct facial anomalies, congenital heart defects, limb and skeletal anomalies, and short stature. Renal anomalies have been reported in a few cases of KS, but to our knowledge, hepatic anomalies have not. Here, we document two cases of KS requiring liver or kidney transplantation: one with severe hepatic and renal anomalies and one with severe renal anomalies. Both cases had the characteristic facial appearance of children with KS, postnatal growth deficiency, and developmental delay. At birth, case 1 presented with hypoglycemia, ileal perforation, right hydroureter, and hydronephrosis. The patient subsequently developed hyperbilirubinemia, hepatic abscess, and cholangitis. At age 8 months, he underwent a liver transplant. Hepatic pathology diagnosed neonatal sclerosing cholangitis. Case 2 presented with renal failure at age 6 years. Renal ultrasound study showed markedly dysplastic kidneys requiring transplantation. In addition to characteristic findings of KS, she had coronal synostosis and was shown to have immune deficiency and an autoimmune disorder manifesting as Hashimoto thyroiditis and vitiligo. We conclude: 1) severe hepatic and renal anomalies leading to organ failure can occur in KS; 2) patients with neonatal sclerosing cholangitis should be examined closely for features of KS; 3) coronal synostosis may occur in KS; and 4) immune deficiency and autoimmune disorder can be associated with KS. PMID- 9856565 TI - Identification of an unusual marker chromosome by spectral karyotyping. AB - We ascertained a newborn girl with multiple congenital anomalies including severe hypotonia, cardiovascular defects, hearing loss, central nervous system anomalies, and facial anomalies. The infant died at 12 days. Cytogenetic analysis showed a de novo supernumerary marker chromosome. Fluorescence in situ hybridization (FISH) with a combination of chromosome specific alpha-satellite probes and an all-human centromere probe failed to show hybridization to the marker, indicating that the marker chromosome lacked detectable alpha satellite sequences. Spectral karyotyping (SKY) was performed and showed that the marker was chromosome 15 in origin. This was confirmed by FISH with a 15q specific subtelomerie probe, which showed hybridization to both ends of the marker chromosome. Based on FISH information and G-banding pattern, the marker was determined to be an inverted duplication of 15q25-qter, leading to partial tetrasomy for chromosome 15. Although the marker chromosome lacked detectable centromeric alpha-satellite sequences, it seemed to have a functional centromere as it is mitotically stable. This observation is consistent with previous studies on acentric marker chromosomes, which suggested that the DNA sequence at the breakpoint could function similarly to alpha-satellite sequences once activated through marker formation. PMID- 9856566 TI - Visual impairment due to macular disciform scars in a 20-year-old man with Smith Magenis syndrome: another ophthalmologic complication. AB - We describe a 20-year-old man with Smith-Magenis syndrome and a 46,XY,del(17)(p11.2p11.2) karyotype. The interstitial deletion was confirmed by metaphase analysis using the fluorescent in situ hybridization probe (D17S29) for the Smith-Magenis region. The patient had hypertelorism, exotropia, and high myopia. Examination under anesthesia showed a lacquer crack near the right macula and a disciform scar of the left macula. Six months later, the patient presented with subacute visual loss. Examination demonstrated end-stage macula degeneration with bilateral disciform scars. There was no evidence of retinal detachment. Prior reports of Smith-Magenis syndrome mention telecanthus, ptosis, strabismus, iris anomalies, cataract, microcornea, optic nerve hypoplasia, myopia, retinal detachment, and lattice retinal degeneration. Bilateral macular degeneration has not been reported previously, and it may be an additional ophthalmologic manifestation of Smith-Magenis syndrome, either as a primary manifestation or as a direct consequence of high myopia. PMID- 9856567 TI - Inherited duplication Xq27-qter at Xp22.3 in severely affected males: molecular cytogenetic evaluation and clinical description in three unrelated families. AB - We describe the clinical phenotype in four males from three families with duplication (X)(qter-->q27::p22.3-->qter). This is an unusual duplication of the distal long arm segment, Xq27-qter, onto the distal short arm of the X chromosome at Xp22.3, as shown by fluorescent in situ hybridization analysis with multiple X specific probes. The patients are young male offspring of three unrelated, phenotypically normal carrier women. The affected males have similar clinical manifestations including severe growth retardation and developmental delay, severe axial hypotonia, and minor anomalies. Such clinical similarity in three unrelated families demonstrates that this chromosome abnormality results in a new and distinct clinical phenotype. Replication studies, performed on two of the mothers, provided evidence that inactivation of the abnormal X chromosome permitted the structural abnormality to persist in these families for a generation or more in females without phenotypic expression. PMID- 9856568 TI - Angelman syndrome: are the estimates too low? AB - More than 300 cases of Angelman Syndrome (AS) have been reported. AS is still considered a clinical diagnosis because only approximately 80% of those individuals who meet the clinical criteria will have a maternal deletion of chromosome 15q11-13. Of the reported cases of AS, very few are of adults with AS. We present our findings on 11 adults with AS identified in a long-term residential care facility for persons with severe developmental disabilities. The diagnosis of AS was not recognized at the time of their admission but was established as part of our evaluation. Thus, there may be an underestimate of the true incidence of AS especially in adults with severe developmental disabilities. PMID- 9856569 TI - Frequent loss of imprinting of the H19 and IGF-II genes in ovarian tumors. AB - Several human imprinted genes have been identified and are implicated in genetic diseases and tumorigenesis. We studied alterations of two imprinted genes, the paternally imprinted H19 and maternally imprinted IGF2, in 15 ovarian tumors with various cell types. To know allele-specific expression of the two genes, we analyzed restriction fragment length polymorphisms (RFLPs) at the 3'-untranslated region (UTR) in their cDNA, compared with those in the respective genomic DNA. As a result, biallelic H19 and IGF2 expression was observed in 8 (62%) of 13 informative (heterozygous) ovarian cancers and in 6 of 11 informative cases, respectively. H19 loss of imprinting (LOI) was most frequently observed in malignant serous cystadenocarcinoma (in four of six cases), whereas IGF2 LOI was not common in malignant epithelial cancers because three of six such LOI events occurred in benign mucinous cystadenomas and non-cancerous endometriotic cyst. Our data suggest that the alteration of H19 and IGF2 imprinting plays differential roles in tumorigenesis and progression of ovarian tumors, depending on the tissue type as well as the developmental stage. Our data may argue against tumor suppressor activity of H19 in ovarian cancers. PMID- 9856570 TI - Association of terminal chromosome 1 deletion with sertoli cell-only syndrome. AB - We report on del(1)(q44), developmental delay, cryptorchidism, and seizure disorder in a 19-year-old man. Endocrinologic evaluation showed delayed puberty and elevated gonadotropins. Testicular biopsy was consistent with Sertoli cell only syndrome. The case illustrates a previously an unreported manifestation in males with del(1)(q44), and suggests a link between the development of germinal epithelium and genes in the 1q44 area. PMID- 9856571 TI - Germline PTEN mutation in a family with Cowden syndrome and Bannayan-Riley Ruvalcaba syndrome. AB - Clinical overlap between Cowden disease and Bannayan-Riley-Ruvalcaba syndrome has rarely been described and identical germline mutations in the PTEN gene have been demonstrated in a few families with Cowden disease and some cases of Bannayan Riley-Ruvalcaba syndrome. We report on a mother with Cowden disease and a son with Bannayan-Riley-Ruvalcaba syndrome. Mutation analysis of the PTEN gene demonstrated a heterozygous nonsense mutation R130X in both individuals. This might suggest that Cowden disease and Bannayan-Riley-Ruvalcaba syndrome are one causal entity. PMID- 9856572 TI - Short stature, brachydactyly, nail dysplasia, and mental retardation: further observation of the Tonoki syndrome. AB - We studied a patient with microcephaly, short stature, type B brachydactyly, nail dysplasia, skeletal anomalies, and mental retardation. The mother of the propositus has brachydactyly of thumbs and a similar physiognomy without mental retardation. This appears to be another observation of the Tonoki syndrome, a distinct autosomal dominant or X-linked clinical entity. PMID- 9856573 TI - Point mutation in the MITF gene causing Waardenburg syndrome type II in a three generation Indian family. AB - Waardenburg syndrome (WS) is an autosomal-dominant neural crest cell disorder phenotypically characterized by hearing impairment and disturbance of pigmentation. A presence of dystopia canthorum is indicative of WS type 1, caused by loss of function mutation in the PAX3 gene. In contrast, type 2 WS (WS2) is characterized by normally placed medial canthi and is genetically heterogeneous; mutations in MITF (microphthalmia associated transcription factor) associated with WS2 have been identified in some but not all affected families. Here, we report on a three-generation Indian family with a point mutation in the MITF gene causing WS2. This mutation, initially reported in a Northern European family, creates a stop codon in exon 7 and is predicted to result in a truncated protein lacking the HLH-Zip or Zip structure necessary for normal interaction with its target DNA motif. Comparison of the phenotype between the two families demonstrates a significant difference in pigmentary disturbance of the eye. This family, with the first documented case of two unrelated WS2 families harboring identical mutations, provides additional evidence for the importance of genetic background on the clinical phenotype. PMID- 9856574 TI - Segregation analysis of Parkinson disease. AB - Parkinson disease (PD) is a prevalent movement disorder of unknown cause whose incidence rises with increasing age. Nearly 20% of PD is familial, a small subset of which exhibits autosomal dominant transmission. However, in most families, the inheritance is not clear. To determine the most likely mode of inheritance of PD, we performed complex segregation analyses using kindreds of 136 PD patients randomly ascertained from a clinic population. The hypotheses of a nontransmissible environmental factor, no major gene or type (sporadic), and all Mendelian inheritance (dominant, recessive, additive, decreasing) were rejected (P <0.001). Familial clustering of PD in this data set is best explained by a rare familial factor which a) is transmitted in a nonMendelian fashion, and b) influences the age at onset of PD. If confirmed, our results have immediate implications in gene-mapping studies which often search for genes that behave in a Mendelian fashion that affect susceptibility rather than age at onset and long term implications in understanding the pathogenesis of PD. PMID- 9856575 TI - Centromeric DNA break in a 10;16 reciprocal translocation associated with trisomy 16 confined placental mosaicism and maternal uniparental disomy for chromosome 16. AB - Stable centromeric breakage in non-acrocentric chromosomes and balanced reciprocal translocation mosaicism are both rare events. We studied a family in which the mother had mosaicism for a balanced reciprocal translocation between chromosomes 10 and 16 which was associated with a break in chromosome 16 centromere alpha-satellite DNA ?146,XX,t(10;16)(q11.2;q11.1) [29]/46,XX[25]?. The derivative chromosome 16 contained only a very small amount of 16 alpha-satellite DNA while the derivative 10 contained all of the 10 alpha-satellite DNA as well as a large amount of the 16 alpha-satellite DNA. The same translocation was present in all cells in her son who was found prenatally to have trisomy 16 mosaicism ?46,XY,t(10;16) (q11.2;q11.1)mat[22]/47,idem,+16[4]?. Trisomy 16 cells were subsequently determined to be confined to the placenta. DNA polymorphism analyses in the family demonstrated maternal uniparental disomy for chromosome 16 in the diploid child. The child, at age 7 months, had minor facial anomalies similar to a previously reported case of maternal uniparental disomy for chromosome 16. In addition to illustrating several rare events, this family further demonstrated that substantial deletion of the centromeric alpha-satellite DNA does not impair centromere function and both mitotic and meiotic stability are retained in such cases. PMID- 9856576 TI - Lethal osteosclerotic osteochondrodysplasia with platyspondyly, metaphyseal widening, and intracellular inclusions in sibs. AB - We report on a previously undescribed form of lethal osteosclerotic skeletal dysplasia in sibs from nonconsanguineous parents. Radiographic findings included increased density in the base of the skull, clavicles, vertebrae, ribs, and the metaphyseal regions of the long bones. There was midface hypoplasia, a large anterior fontanel, micrognathia, and hypoplastic, wafer-thin vertebrae. The clavicles, ribs, metacarpals, metatarsals, and phalanges were especially thickened and widened. The long bones were shortened with flared metaphyses. Chondroosseous morphology of resting cartilage and growth plate was relatively normal, but there was hypercellular cortical and trabecular bone, and marrow fibrosis. Ultrastructurally, the resting chondrocytes, osteoblasts, and nonhematopoietic marrow cells had dilated rough endoplasmic reticulum (inclusion bodies). The radiographic and morphologic characteristics in this case are unique and differ from those seen in other previously reported lethal osteosclerotic skeletal dysplasias. PMID- 9856577 TI - New autosomal recessive syndrome of severe microcephaly and skeletal anomalies including posterior rib-gap defects. AB - We describe two female fetuses conceived by a nonconsanguineous couple. The pregnancies were interrupted at 31 and 26 weeks of gestation, respectively, because of severe microcephaly. Postmortem X-ray and autopsy studies showed in both fetuses: 1) severe intrauterine growth retardation; 2) facial anomalies characterized by severe microcephaly, sloping forehead, low set and posteriorly angulated ears, prominent eyes, down-slanting palpebral fissures, large nose, small mouth with full lips, and mild microretrognathia; 3) severe brain hypoplasia that was more pronounced in the second fetus; 4) severe rib hypoplasia with posterior rib-gap defects and in case 2 hypoplasia of several bones (right clavicle, right radius and ulna, several phalanges of hands and feet); 5) contracture at large joints. No other visceral malformations were observed, and chromosomes were normal in patient 2 and parents. This phenotype has some similarities with different syndromic entities but an identical malformation syndrome seems not to have been described previously. Autosomal recessive inheritance is the most likely cause of this putative "new syndrome." PMID- 9856578 TI - Hypomelanosis of Ito: a symptom, not a syndrome. PMID- 9856579 TI - Familial occurrence of a del(Xp-) chromosome: pitfall in karyotype/phenotype correlation. PMID- 9856580 TI - Re: Metacarpophalangeal analysis in Crouzon syndrome. PMID- 9856581 TI - A further case of choanal atresia in the deletion (9p) syndrome. PMID- 9856582 TI - Plasma copolymer surfaces of acrylic acid/1,7 octadiene: surface characterisation and the attachment of ROS 17/2.8 osteoblast-like cells. AB - The purpose of this study was: (a) to examine the effect of plasma-gas composition on plasma polymer oxygen/carbon (O/C) ratio, functional group composition and stability in water, and then (b) to examine cell attachment to surfaces containing different concentrations of O/C and functional groups. Oxygen functionalised surfaces were deposited by means of the plasma copolymerisation of acrylic acid/1,7-octadiene. The use of a diluent hydrocarbon allowed the deposition of surfaces with a range of O/C concentrations. Plasma copolymer surfaces were characterised by X-ray photoelectron spectroscopy (XPS). Changes in functional group composition with % acrylic acid monomer and the non-dispersive and dispersive parts of the surface energy of these plasma copolymers were measured. The solubility of the plasma copolymers was assessed by means of XPS. The degree of attachment of ROS 17/2.8 osteoblast-like cells to plasma copolymer surfaces deemed to be 'stable' in aqueous medium was measured. Tissue culture polystyrene (TCPS) was included as a control. Attachment was found to be greatest to the plasma copolymer surface with an O/C of 0.11. This surface had a carboxylic acid concentration of ca. 3%. Attachment did not correlate with increased surface wettability (i.e. the non-dispersive component of the surface energy). PMID- 9856583 TI - A new model to assess staphylococcal adhesion to intraocular lenses under in vitro flow conditions. AB - Adhesion of staphylococcal cells to intraocular lenses coated with heparin was studied under in vitro flow conditions (280 microl min(-1)) at 37 degrees C. The intraocular lenses were incubated with human cerebrospinal fluid for 1 h or with cerebrospinal fluid including 0.50% plasma for 12 h, prior to bacterial challenge. Two strains of Staphylococcus epidermidis selected for this study, were isolated from biomaterial-associated infections. Bacterial adhesion was quantitated by bioluminescence and visualized by fluorescence microscopy of acridine orange stained bacteria. Surface coating with heparin significantly decreased bacterial adhesion of both strains after incubation with cerebrospinal fluid including 0.50% plasma for 12 h (p = 0.0209). However, no difference in bacterial adhesion was obtained between intraocular lenses with and without heparin, after incubation with cerebrospinal fluid for 1 h (p = 0.327). Microscopy showed that more bacteria were present on intraocular lenses without heparin than on intraocular lenses with heparin. The results show that preincubation with a proteinaceous fluid influences subsequent bacterial adhesion to the polymer surface. The results suggest that IOLs with heparin coating may be less prone to bacterial adhesion under perfusion conditions after surface conditioning in human CSF with 0.50% plasma and a preincubation period of 12 h. Heparin coating might be a valuable tool to decrease implant-associated bacterial endophthalmitis. PMID- 9856584 TI - A quantitative study of the sintering and mechanical properties of hydroxyapatite/phosphate glass composites. AB - Previous work has shown that small additions of a phosphate glass (CaO-P2O5) can significantly enhance the sinterability and strength of hydroxyapatite. However, there are no quantitative phase analyses available for these materials which would provide indicators of biocompatibility and resorbability. Similarly, there is little information available about the mechanical properties, especially with high glass additions. In this study, the effects of sintering hydroxyapatite with phosphate glass additions of 2.5, 5, 10, 25, and 50 wt.% are quantified. Each composition was sintered over a range of temperatures, and quantitative phase analysis was carried out using XRD. In addition, the microstructures were studied using RLOM and SEM, and mechanical properties (Vickers hardness, KIC, and MOR) measured. These results may be used to indicate which compositions and processing conditions may provide materials suitable for use in hard tissue replacement. Composites containing up to 10 wt.% glass additions formed dense HA/TCP composite materials possessing flexural strength and fracture toughness values up to 200% those of pure HA. The HA/TCP ratio was strongly dependent on the percentage glass addition. Higher glass additions resulted in composites containing beta-TCP together with large amounts of alpha- or beta-calcium pyrophosphate, and having similar mechanical strengths to pure HA. PMID- 9856585 TI - High-strength, ultra-thin and fiber-reinforced pHEMA artificial skin. AB - Membranes of pHEMA-based composites were manufactured by adding various kinds of weaved and knitted fabrics and fibers into a deionized water solution of HEMA monomer, EGDMA cross-linker and BIE initiator, and followed by polymerization under ultraviolet radiation. By varying the amount of initial water addition (IWA), the dimensional change of pHEMA matrix from the newly fabricated state to the eventually swollen state could be adjusted to reduce the swellability mismatch with the fabrics and the possibility of the swollen membranes becoming folded and curled was avoided. Mechanical properties of the fiber-reinforced pHEMA composites, including yielding strength, maximum strength, Young's modulus and elongation at break, are improved evidently depending on the mechanical characteristics of additives applied. The involvement of fabrics and fibers in the soft pHEMA matrix also provides an alternative of making the ultra-thin membranes to overcome the problem of easily being torn during handling. In addition, some of these membranes also exhibit an improvement in water transmission rate. PMID- 9856586 TI - Selective plasminogen binding: cysteinyl-lysine-dextran protein interactions. AB - The dipeptide, L-cysteine-L-lysine, when coupled to a dextran matrix demonstrates a much higher binding of Glu-plasminogen than dextran alone. Plasminogen binds to the dipeptide coupled matrix with a high affinity and binding capacity and is retained by the surface for long periods. In comparison, other proteins such as bovine serum albumin, alpha2-antiplasmin, tPA, fibrinogen, and plasmin are not retained by the material demonstrating that the L-cysteine-L-lysine-dextran is highly selective for Glu-plasminogen. While fibrinogen alone does not bind to the surface, fibrinogen on surfaces to which plasminogen has been pre-adsorbed does demonstrate adsorption interactions. The nature of the interactions depend on the amount of plasminogen bound. Fibrinogen in contact with high concentrations of adsorbed plasminogen causes an increase in the desorption rate of the surface bound protein; if low amounts of plasminogen are initially bound, fibrinogen exposure causes an increase in total protein adsorbed by the surface. A higher amount of protein adsorbs from a mixed plasma solution. This does not appear to inhibit the binding of plasminogen, but rather may actually facilitate more plasminogen binding than is the case of plasminogen alone. The dipeptide coupled to the dextran matrix appears to be a preferred substrate for plasminogen binding compared to free lysine and epsilon-amino caproic acid. PMID- 9856587 TI - Some characteristics of hydroxylapatite powder particles after plasma spraying. AB - We have studied the particles of hydroxylapatite (HA) powder, the particles after plasma spraying, their distribution on substrate surface and their condition after transfer through the plasma torch. Mean particle size of HA powders was as follows: HA-A: 3.8 microm, HA-B 88.2 microm. The area of HA coating after plasma spraying, when the torch had a constant position against the substrate surface, shows two characteristic zones: the central part of coating formed mainly from deformed particles and the marginal part of coating with small non-deformed particles. These small non-deformed particles can be found in all zones of the coating and together with greater non-deformed particles and partially deformed particles will unfavourably affect the adhesive and cohesive strength of the coating and its porosity. The maximum diameter of the molten (spherical) particles in the conditions of Ar + H2 plasma, output P = 24 kW was: DA = 25 microm (HA-A) and DB = 65 microm (HA-B). The intervals of dimensions in which most of molten particles occurred were HA-A: 0-15 microm (98%), HA-B: 5-35 microm (84%). From comparison of HA-A and HA-B powders it can be concluded that the transport of HA-A powder was not continuous, the amount of molten HA-A particles was considerably greater (90%) than that of HA-B powder (63%). Phase decomposition and also solubility of HA-A powder (at in vitro tests) was greater. If we consider transport of particles, their melting, splitting and spraying efficiency, the suitable size of HA powder particles for the given spraying conditions is somewhere between the size of HA-A and HA-B particles--let us say- in the interval from 20 to 60 microm. PMID- 9856588 TI - Surface roughness of debonded straight-tapered stems in cemented THA reduces subsidence but not cement damage. AB - Although stress analyses have shown that the mechanical endurance of cemented femoral THA reconstructions is served by stems that firmly bond to their cement mantles, retrieval studies suggest that this may be difficult to achieve. Clinical studies with roentgen stereophotogrammetric analyses have shown that stems may gradually debond from their cement mantle. Accepting the fact that stem debonding is unavoidable, stem subsidence and cement stresses can be reduced by increasing stem-cement friction, as indicated by finite element stress analyses. Hence, it can be hypothesized that debonded stems with high surface roughness values would damage the cement mantle to a lesser extent as compared to polished ones. To confirm this hypothesis, tapered stems with polished and rough surface finishes were implanted in cement mantles and cyclically loaded for 1.7 million times. It was investigated how surface roughness affected the damage in the cement mantle, and how it was related to prosthetic subsidence. The polished taper subsided considerably more than the rough one (630 vs. 270 microm at the end of the experiments). In addition, it was found that the polished taper displayed step-wise subsidence, which is probably due to the interaction of stick slip processes at the interface, associated with creep of the acrylic cement. The rough taper subsided monotonously. Scanning Electron Microscopic (SEM) analysis of the taper-cement structures showed that the rough taper was completely debonded from the cement mantle, creating a gap at the interface, and that many large cement cracks and particles were created. Around the polished taper, only a few cracks were found and the taper-cement interface seemed undamaged. It was concluded that an increased surface roughness does not necessarily lead to a reduction in cement damage. On the contrary, compared to polished ones, debonded rough stems may produce more cement cracks and acrylic cement debris, and provide routes to transport these wear products. Hence, after failure of the stem-cement interface, straight-tapered stems with an increased surface roughness accelerate the failure process due to inferior fail-safe features. Consequently, in vivo subsidence patterns at the stem-cement interface should be considered in combination with the surface finish of the implant. An amount of post-operative subsidence of a rough stem may be much more damaging for the reconstruction than the same amount for a polished stem. PMID- 9856589 TI - In vitro sorption and desorption of basic fibroblast growth factor from biodegradable hydrogels. AB - In vitro interaction of basic fibroblast growth factor (bFGF) with biodegradable gelatin hydrogels was investigated, focusing on its sorption into the hydrogels and desorption from them. Basic bFGF was sorbed to the hydrogel of acidic gelatin with an isoelectric point (IEP) of 5.0 over time at 4 degrees C, in contrast to that of basic gelatin with an IEP of 9.0 and type I collagen. The bFGF sorption was almost independent of the sorption temperature except for 4 degrees C and the hydrogel water content. Fluorescent microscopic observation revealed that bFGF was sorbed into the interior of the acidic gelatin hydrogel. The binding molar ratio of bFGF to the acidic gelatin was around 1.0. The bFGF sorption to the acidic gelatin hydrogel increased when gelatin was further carboxylated. bFGF was sorbed into the acidic gelatin hydrogel more slowly than into the poly(acrylic acid) (PAAc) hydrogel, probably because of the lower density of negative charge of gelatin. The bFGF sorption decreased with an increase in solution ionic strength, indicating that an electrostatic interaction was the main driving force for bFGF sorption to the acidic gelatin hydrogel. However, even at higher ionic strengths of solution, the sorbed bFGF was not desorbed from the acidic gelatin hydrogel, in contrast to the PAAc hydrogel. PMID- 9856590 TI - DNA image cytometry and Ag-NORs-staining application in biocompatibility studies on human osteoblast cells in vitro. AB - We report here the study of the biocompatibility of a bone graft material, the Pyrost, using a previously established in vitro model of human osteoblasts. The effect of this material on cell proliferation was evaluated by the MTS assay. Results indicated the absolute absence of cytotoxic or cytostatic effect of Pyrost on cultured osteoblasts. Viability rate was more than 90% in cells cultured with the material compared to the control. Morphological analysis, undertaken by scanning electron microscopy showed a good adhesion and a spreading of osteoblasts in contact with the material that was colonized by cultured cells. In the second part of this work, we have introduced two methods as complementary biocompatibility tests: DNA image cytometry and interphase Ag-NORs quantification. DNA content was measured in cells cultured with or without Pyrost for 3, 9, 15 and 30 days. The determination of DNA indicated that the majority of osteoblasts population was diploid without aneuploidy. The DNA index and cell distribution profile in DNA histograms were similar in all cell populations. The Ag-NORs amount was used as a parameter for cell kinetic evaluation. We have measured the Ag-NORs index like DNA quantification. The proliferation rate, evaluated by Ag-NORs counts in osteoblasts cultured with or without the material, was identical. However, a decrease in Ag-NORs index was observed from day 3 to day 15 of incubation. These results showed a satisfactory biocompatibility of the Pyrost in human osteoblasts culture. The material did not alter cell viability and had no inducing effect either on proliferation rate or on cell ploidy as demonstrated by DNA image cytometry and Ag-NORs proteins staining. PMID- 9856591 TI - Fusion of macrophages on an implant surface is associated with down-regulated expression of ligands for galectin-1 and -3 in the rat. AB - Galectins have a wide range of biological activities which are elicited by binding to appropriate glycoligands. Besides regulation of the expression of the galectins the extent of the presence of suitable binding sites will be relevant to infer the cellular responsiveness to this class of sugar receptors. Thus ligand presentation requires monitoring by the tissue lectin. We demonstrate the expression of galectin-3 by macrophages and foreign-body giant multinucleate cells colonizing a cellophane implant in the rat by the A1D6 monoclonal antibody. The extents of ligand presence are visualized in the same cells by biotinylated galectin-3 and also by galectin-1 which is produced by diverse mammalian cell types and widely distributed. Labeled mistletoe (VAA) and tomato (LEA) lectins are used as tools to assess the degree of similarity of the binding profile between endogenous and exogenous proteins. The presentation of alpha-galactosides is monitored with a natural immunoglobulin G subfraction obtained by two consecutive affinity chromatography steps. The binding of labeled galectins and plant lectins was significantly lower to foreign-body giant multinucleate cells than to mononuclear macrophages. The application of the alpha-galactoside specific probe yielded no significant staining. The potential problem of epitope accessibility could be excluded by the concomitant positivity obtained with an IgG subfraction with selectivity to beta-galactosides also obtained by affinity chromatography. These results provide no evidence for a role of alpha galactosides for the binding of galectins in the rat macrophages colonizing the implant. The reduced level of expression of glycoligands for galectin-1 and -3 in foreign-body giant multinucleate cells in contrast with the mononuclear macrophages suggests an inhibitory influence of macrophage fusion on the expression of galectin-reactive molecules. PMID- 9856592 TI - Investigation into the effects of proteins and lipids on the frictional properties of articular cartilage. AB - The aim of this study was to investigate the effects of the proteins and the lipids on the frictional properties of articular cartilage in the mixed and boundary lubricating regimes. Bovine articular cartilage that had been treated to remove the lipids, and cartilage that had been treated to remove the proteins, from the surface layer were studied in order to investigate differences in their lubricating abilities. Tests were carried out on a sliding friction rig under contact pressures of 0.5 and 4 MPa. The results of the friction tests carried out under the 4 MPa contact pressure showed a slight increase in friction coefficient for both the lipid and the protein deficient cartilage when compared with the control cartilage. These differences were more apparent at the shorter loading times but none of the differences were found to be statistically significant. The tests at the lower contact pressure of 0.5 MPa showed that removing the lipids by washing the surface with detergent increased the friction coefficient at the shorter loading times but had no effect at the longer loading times. Digesting the cartilage with trypsin to remove the proteins from the surface layer had no effect on the friction coefficient at the shorter loading times but reduced the friction coefficient at the longer loading times. Both these results were confirmed with specimens that had both the lipids and the proteins removed from the surface layer. PMID- 9856593 TI - Alternation in the flutter wave morphology during radiofrequency catheter ablation for common atrial flutter. AB - A linear lesion created at the right atrial isthmus by radiofrequency current application can successfully eliminate common atrial flutter (AF). The mechanism of unsuccessful cases has not yet been well delineated. This study sought to investigate the cause of unsuccessful cases of radiofrequency catheter ablation of AF. Sixty-six patients with refractory common AF were referred for radiofrequency catheter ablation. Radiofrequency current was applied to the right atrial isthmus between the inferior vena cava and tricuspid annulus or between the coronary sinus orifice and tricuspid annulus. In 5 (8%) of the 66 patients, a morphological change of the flutter wave was observed in the 12-lead ECG concomitant with the change of the atrial excitation sequence during the delivery of radiofrequency energy without the termination of atrial flutter. In 8 (12%) patients, the morphology of the new AF wave, which was provoked electrically after the termination of the original AF, was different, and the average flutter cycle length also differed in 3 cases (2%). The results of radiofrequency application could be misinterpreted as unsuccessful when the occurrence of another, different type of AF has been overlooked following the elimination of the original AF during the radiofrequency catheter ablation procedure. It is possible that the flutter circuit can take an alternative pathway despite the complete conduction block at the right atrial isthmus. PMID- 9856594 TI - Comparison of thrombolytic therapies with mutant tPA (lanoteplase/SUN9216) and recombinant tPA (alteplase) for acute myocardial infarction. AB - The fibrinolytic capacity of patients with acute myocardial infarction (AMI) is known to be impaired. The primary regulatory element of the fibrinolytic system is plasminogen activator inhibitor (PAI). It has been previously observed that there are 2 peaks in the plasma PAI level of AMI patients at 4h and 16h after thrombolytic therapy with recombinant tissue plasminogen activator (rtPA). Lanoteplase/SUN9216 is a mutant tPA with a biological half-life longer than that of rtPA. Thrombolytic therapy with mutant tPA or rtPA was carried out consecutively in 21 patients with AMI (8 patients as the mutant tPA group, and 13 patients as the rtPA group). The recanalization time of the mutant tPA group was significantly faster than that of the rtPA group (16.1 +/- 3.9 min vs 39.6 +/- 4.8 min, p<0.01). The PAI activity at 4h after the initiation of thrombolysis was significantly lower in the mutant tPA group than in the rtPA group (8.74 +/- 5.46IU/L vs 26.74 +/- 3.35 IU/L, p<0.01). There was a one mild peak in serial plasma PAI activity levels 24h after the initiation of thrombolysis. The results suggest that thrombolytic therapy with mutant tPA reduced the impairment of fibrinolytic capacity. The mutant tPA gives faster recanalization and lower PAI activity after successful thrombolysis, compared with rtPA. PMID- 9856595 TI - Disopyramide: a promising new approach to the medical treatment of the hypercyanotic spell complicating tetralogy of Fallot. AB - Under continuous ECG and oxygen saturation (SpO2) monitoring, the following measurements were taken by Doppler echocardiography in 6 consecutive patients with tetralogy of Fallot (TF) before and after intravenous administration of disopyramide (2mg/kg): left ventricular shortening fraction (LVSF); peak velocities in the right ventricular outflow tract (RVOT); diastolic and systolic internal diameters of the right ventricular outflow tract (dRVOT, sRVOT); and systolic blood pressure. SpO2 increased (p<0.01) from 78 to 98 (89 +/- 7, mean +/ standard deviation)% to 86-99 (94 +/- 5)%. LVSF decreased (p<0.05) from 0.34 0.56 (0.42 +/- 0.08) to 0.22-0.54 (0.33 +/- 0.13). The systolic blood pressure fell slightly (p<0.05) from 68-92 (79 +/- 8) to 64-92 (71 +/- 11)mmHg. The sRVOT increased (p<0.05) from 2.1-4.8 (2.7 +/- 1.5)mm to 3.0-8.1 (4.9 +/- 2.4)mm, while RVOT peak velocity decreased (p<0.05) from 2.20-4.88 (3.70 +/- 0.97)m/sec to 2.05 4.07 (2.92 +/- 0.72)m/sec. Disopyramide alleviates hypoxia in patients of TF through its negative inotropic action on right ventricular outflow obstruction. PMID- 9856596 TI - Solitary papillary muscle hypertrophy as a possible form of hypertrophic cardiomyopathy. AB - Patients can present with hypertrophied papillary muscles in the left ventricle, even without hypertrophy in other segments, and they have electrocardiographic (ECG) abnormalities suggestive of hypertrophic cardiomyopathy (HCM). This study was performed to evaluate whether the solitary papillary muscle hypertrophy was related to HCM. By analyzing 6731 echocardiographic studies between 1990 and 1994, the incidence of patients with papillary muscle hypertrophy was retrospectively examined, as well as the ECG features and family history related to HCM in these patients. After the normal size of the anterolateral and posteromedial papillary muscles was obtained from echocardiographic studies in 40 healthy subjects (0.7 +/- 0.2 cm for each of the vertical and horizontal axis), papillary muscle hypertrophy was defined as follows: either the vertical or horizontal diameter of at least one of the 2 papillary muscles was more than 1.1 cm (mean+2SD in the normal subjects). Using this definition, 29 patients with papillary muscle hypertrophy were identified, of whom 14 (48%) showed high voltage QRS complexes, 10 (34%) showed T wave inversion, and 6 (21%) showed abnormal Q waves. Ten patients (34%) had a family history of HCM. In 2 patients that were followed for 18 and 11 years, respectively, the voltages of the QRS complexes and inverted T waves progressed with the hypertrophy of the papillary muscle. These findings suggest that solitary papillary muscle hypertrophy is related to HCM and that papillary muscle hypertrophy is a newly identified subtype of or an early form of HCM. PMID- 9856597 TI - Functional evaluation of the bileaflet mechanical valve in the aortic position using dobutamine-stress echocardiography: is a 23-mm prosthetic valve adequately large enough? AB - The issue of valve prosthesis-patient mismatch in small annular patients is still controversial. The hemodynamic function of bileaflet mechanical valves in the aortic position was examined using dobutamine-stress echocardiography. Forty-four patients were enrolled in the study and divided into 5 groups, according to prosthesis size, from 21 mm to 29 mm. The aortic peak pressure gradient (APG) increased significantly in all groups with dobutamine-stress and exceeded 50 mmHg in 83% of the 21-mm group, in 64% of the 23-mm group, and in 33% of the 25-mm group. The APG even exceeded 80 mmHg in 22% and 18% of the 21-mm and 23-mm groups, respectively. In these cases, the potential of 'valve prosthesis-patient mismatch' was considered. From the relationship between the APG and the prosthesis valve area index (VAI), 'critical VAIs' were found where patients were likely to enter the 'mismatch' status; that is, 1.22 and 1.77 cm2/m2, respectively, for the 5 and 10 microg/kg per min stages of dobutamine stress. This critical VAI range is useful in predicting the 'mismatch' patients preoperatively. Alternative procedures or prostheses should then be selected for them. PMID- 9856598 TI - Effect of amezinium metilsulfate on the finger skin vasoconstrictor response to cold stimulation and venoconstrictor response to noradrenaline. AB - Orthostatic hypotension can be caused by an inadequate vasoconstrictor response. The effects of amezinium on vasoconstrictor response to sympathetic stimulation and to exogenous noradrenaline were investigated and compared with those of midodrine. In 8 healthy men, the following experiments were performed after a single oral dose of 10mg of amezinium, 2mg of midodrine or a placebo. First, finger-tip blood flow (FTBF) was recorded using a laser Doppler flowmeter before and during the contralateral hand cooling and a reduction ratio of FTBF was calculated as an index of the vasoconstrictor response. Second, dose-response curves to increasing doses (1-512ng/min) of noradrenaline infused locally to the dorsal hand vein were determined using a linear variable differential transformer. The reduction ratio of FTBF was significantly increased (p<0.05) by amezimium [placebo, 75.9 +/- 9.8(mean +/- SD)%; amezinium, 85.1 +/- 7.9%; midodrine, 78.1 +/- 9.3%]. The infusion rate of noradrenaline producing a half maximum venoconstriction was significantly decreased (p<0.05) by amezinium (placebo, 40.6 +/- 33.9 ng/min; amezinium, 21.0 +/- 21.3 ng/min; midodrine, 33.2 +/- 31.5 ng/min). These findings indicate that amezinium increases the vasoconstrictor response to sympathetic stimulation and to noradrenaline in normal subjects, and this mechanism might contribute to the improvement by amezinium of the symptoms of orthostatic hypotension. PMID- 9856599 TI - Application of mechanical restitution: variation of inotropic effects of vagal stimulation or verapamil administration during irregular cardiac rhythm. AB - The effects of verapamil administration or vagal stimulation on the mechanical restitution curve (MRC) were studied in order to better understand the modulation of left ventricular (LV) function by interventions that lower the ventricular rate of atrial fibrillation. The MRC and the postextrasystolic MRC were obtained in 11 dogs using peak single beat elastance (Emax). The MRC was fitted by a monoexponential curve. Vagal stimulation or verapamil administration decreased the peak of the MRC and right-shifted the MRC. The postextrasystolic MRC was located upward compared with the control MRC, and was shifted downward by vagal stimulation or verapamil administration. If interventions having a negative inotropic effect effectively slow a rapid heart rate, the net effect of the ventricular contractile state may not always be negative. It was concluded that the MRC is useful in understanding LV contractility during irregular rhythm, especially when assessing the net effect of the negative dromotropic and inotropic action of antiarrhythmic drugs. PMID- 9856600 TI - Expression of cell adhesion molecules and the appearance of adherent leukocytes on the left atrial endothelium with atrial fibrillation: rabbit experimental model. AB - To assess the expression of cell adhesion molecules and the appearance of leukocytes adhering to the left atrial endothelium with atrial fibrillation (AF), 10 Japanese white rabbits were anesthetized and 3 pacing leads were placed in the right atrium. For the AF model, the right atrium was stimulated by electrical pacing (the stimulation frequency of each lead being adjusted to different intervals) for 8h while the control model was subjected to a sham operation without atrial stimulation. The left atrial appendage was excised from the heart and examined immunohistochemically. P-selectin staining of the endothelium in both models was linear and regional, and intracellular adhesion molecule-1 (ICAM 1) in the AF model was confined to leukocytes and endothelial cells with adherent leukocytes. The expression of P-selectin (p<0.05) and the appearance of positively ICAM-1 stained adherent leukocytes (p<0.05) were significantly greater in the AF model than in the control model. In conclusion, AF could regulate the expression of at least 2 critical adhesion molecules, P-selectin and ICAM-1, and the appearance of adherent leukocytes; suggesting that these molecules may play an important role in left atrial thrombus formation with AF. Although anticoagulant therapy has generally been carried out with warfarin in AF patients, neutralizing antibodies to cell adhesion molecules should be tried to prevent thromboembolic complications. PMID- 9856601 TI - Decrease in the high frequency QRS components depending on the local conduction delay. AB - The high frequency components contained in the QRS complex (HF-QRS) are a powerful indicator for the risk of sudden cardiac death. However, it is controversial whether conduction delay increases or decreases the HF-QRS. In 21 anesthetized, open-chest dogs, the right atrium was constantly paced. A cannula was inserted into the left anterior descending artery and flecainide, lidocaine or disopyramide was infused to slow the local conduction. Sixty unipolar electrograms were recorded from the entire ventricular surface and were signal averaged. Data were filtered (30-250 Hz) by using fast-Fourier transform. The HF QRS was calculated by integrating the filtered QRS signal. Activation time (AT; dV/dt minimum) was delayed and the HF-QRS was reduced in the area perfused by flecainide, lidocaine or disopyramide. The percent increase in AT closely correlated the percentage decrease in the HF-QRS; the correlation coefficients were 0.75, 0.83 and 0.76 for flecainide, lidocaine and disopyramide infusion, respectively, (p<0.001). Decrease in the HF-QRS linearly correlated with the local conduction delay. This study proved that conduction delay decreases the HF QRS, and that the HF-QRS is a potent indicator of disturbed local conduction. PMID- 9856602 TI - A premenopausal woman presenting with acute myocardial infarction of three different coronary vessels within 1 year: role of lipoprotein(a). AB - A premenopausal female patient presented with acute myocardial infarction of 3 different coronary vessels at different times within 1 year. These events were caused not by restenotic lesions after balloon angioplasty but by new lesions, which were successfully treated by primary angioplasty. Although she had a history of hypertension, type IIB hyperlipidemia, and diabetes, they had been well-controlled on medication. An elevated serum lipoprotein(a) level may have played a role in this rapid angiographic progression. PMID- 9856604 TI - Treatment of hypertension on the basis of home measurements: a case report. AB - Should antihypertensive therapy be based on home blood-pressure monitoring in hypertensive patients with marked white-coat phenomenon? A 53-year-old hypertensive woman with marked white-coat phenomenon had been treated on the basis of blood pressure at home without measurements of clinic blood pressure for 5 years. The target of her treatment was changed to clinic blood pressure because of marked electrocardiographic abnormalities, which were subsequently normalized by the treatment given for the office hypertension. Thus, the experience of this case suggests that treatment of cases of hypertension with white-coat phenomenon should not be based only on home recordings. PMID- 9856603 TI - Two cases of hypertrophic cardiomyopathy with coronary vasospasm. AB - Chest pain in patients with hypertrophic cardiomyopathy seems to be caused by relative myocardial ischemia due to the left ventricular outflow pressure gradient and myocardial hypertrophy. However, in 2 cases of hypertrophic cardiomyopathy chest pain was associated with coronary vasospasm. Thus, chest pain in these cases was decreased not by a beta-blocker but by isosorbide dinitrate and a calcium antagonist. Because beta-blockers are commonly used for hypertrophic obstructive cardiomyopathy and chest pain may be aggravated by beta blockers in patients with coronary vasospasm, a combination of beta-blocker, isosorbide dinitrate and calcium antagonist was necessary for this hypertrophic cardiomyopathy with variant angina. PMID- 9856605 TI - Improved cardiac function after catheter ablation in a patient with type B Wolff Parkinson-White syndrome with an old myocardial infarction. AB - A 67-year-old man was admitted to hospital for the treatment of exertional dyspnea. He suffered from congestive heart failure due to an old inferior myocardial infarction with type B Wolff-Parkinson-White syndrome. Asynchronous wall motion caused by pre-excitation through a right-side bypass tract caused his cardiac function to deteriorate. Catheter ablation of the bypass tract increased the ejection fraction, and improved his symptoms, prior to surgical revascularization. PMID- 9856607 TI - Polarization potentials causing pacemaker oversensing. AB - Pacemaker oversensing with inappropriate inhibition is commonly caused by sensing of either T-waves or polarization potentials, which are difficult to distinguish from one another. Decisive evidence, obtained from telemetered electrograms and subthreshold pacing, is presented to support sensing of the polarization potential alone and the exclusion of the T-wave as the cause of oversensing in a patient. PMID- 9856606 TI - A case of malignant lymphoma with diastolic heart failure. AB - In patients with malignant lymphoma, the first signs and symptoms are frequently noncardiac and clinical manifestations of cardiac involvement are often nonspecific. This case report presents a patient with malignant lymphoma whose first manifestation was characteristic of heart failure, mainly due to diastolic dysfunction, and whose postmortem examination revealed massive myocardial invasion. PMID- 9856608 TI - High-dose immunoglobulin G therapy for fulminant myocarditis. AB - The cardiac function of an adult patient with fulminant myocarditis requiring a mechanical circulatory support was improved, along with the suppressed release of inflammatory mediators, after intravenous infusion with high-dose immunoglobulin. This therapy may have immunomodulatory effects and serve as a potential adjunctive therapy for patients with fulminant myocarditis. PMID- 9856609 TI - The high cost of spending less on care. PMID- 9856610 TI - Equitable peer review. PMID- 9856611 TI - Management of chronic pain among elderly patients. PMID- 9856612 TI - Is managed care managing not to care? PMID- 9856613 TI - Mania as spiritual emergency. PMID- 9856614 TI - Costs of employee behavioral health care by diagnosis. PMID- 9856615 TI - Web sites worth watching. PMID- 9856616 TI - How a managed behavioral health care carve-out plan affected spending for episodes of treatment. AB - OBJECTIVE: This study examined the impact on spending for episodes of mental health and substance abuse treatment of a managed behavioral health care carve out program implemented by the Massachusetts Group Insurance Commission in July 1993. METHODS: Episodes of mental health and substance abuse treatment were defined using claims and enrollment data from before and after the carve-out implementation. Regression models were used to compare spending per episode for different types of episodes of mental health and substance abuse care: those involving care provided only in an inpatient facility (that is, inpatient care or partial hospitalization), those involving both inpatient-facility and outpatient care, and those involving only outpatient care. RESULTS: Adoption of the carve out plan was associated with a large decrease in spending per episode across all three episode types, particularly for episodes involving inpatient-facility care. The decrease was 54 percent for inpatient-facility-only episodes, 46 percent for combined inpatient facility and outpatient episodes, and 21 percent for outpatient-only episodes. The decrease in spending per episode was larger for episodes involving a diagnosis of either unipolar depression or substance dependence. CONCLUSIONS: The findings suggest that spending per episode of mental health and substance abuse treatment may drop substantially after a carve-out is implemented. Individuals with a diagnosis of either unipolar depression or substance dependence seem to be disproportionately affected. It appears that even weak financial incentives placed on the managed behavioral health care vendor can result in dramatic changes in spending patterns for episodes of mental health and substance abuse treatment. PMID- 9856617 TI - Treatment of major depression before and after implementation of a behavioral health carve-out plan. AB - OBJECTIVE: The study examined utilization, payments, and quality indicators for treatment of major depressive disorder before and after the 1993 implementation of a behavioral health care carve-out plan for Massachusetts state employees who received medical coverage through indemnity plans or preferred provider organizations. METHODS: The sample of 2,259 enrollees with claims for treatment of major depressive disorder was drawn from the group of 39,541 persons continuously enrolled in preferred provider organizations or indemnity plans for fiscal years 1992 to 1995. A subsample of 243 users of inpatient services accounted for 352 admissions. Bivariate tests were used to compare utilization and quality indicators before and after implementation of the carve-out plan. Simple comparisons of current-year dollars were used. RESULTS: The proportion of enrollees with claims for treatment of major depressive disorder increased significantly under the carve-out plan. Inpatient utilization decreased substantially, mostly due to a significantly lower average length of stay (16 days before implementation of the carve-out plan and nine days after). Net inpatient payments fell 71 percent overall, 65 percent per admission, and 40 percent per day. The unadjusted proportion of discharged patients treated for major depressive disorder who were readmitted within 15 and 30 days did not change significantly. The unadjusted proportion of cases receiving follow-up within those time frames increased significantly. CONCLUSIONS: Implementation of a behavioral health carve-out plan may be accompanied by substantial reductions in inpatient utilization and payments for treatment of major depressive disorder. Descriptive findings suggest that such reductions may not have a detrimental impact on readmission and follow-up treatment rates within 30 days. However, this analysis did not control for patient characteristics, used short follow-up periods, and did not include some relevant outcome measures. PMID- 9856618 TI - Overcoming service barriers for homeless persons with serious psychiatric disorders. AB - OBJECTIVE: To help homeless persons with comorbid psychiatric and substance use disorders gain access to community services, in 1993 the Center for Mental Health Services implemented the five-year Access to Community Care and Effective Services, or ACCESS program, in 15 cities. One aim of the program is to encourage collaboration between agencies serving the multiple needs of this population. This study examined the extent of linkages between agencies in the 15 demonstration cities. METHODS: One respondent from each of the 1,060 community based programs in tie 15 cities rated the extent to which his or her agency was linked with each of the other agencies in the local community in 1994 and again in 1996. Overall, there were 20,801 potential pairwise linkages. Linkages were classified into four types: a mutual tie, in which both agencies send and receive clients; a unidirectional tie, in which one agency sends and the other receives; an attempted tie, in which one agency sends but the other agency does not confirm receiving; and an unattempted tie. RESULTS: In 1994 and 1996, of the 20,801 pairs of potential service linkages, about a third were in place, while the remaining two-thirds were absent. Overall, linkages showed a slight but significant increase between 1994 and 1996. More than half of the linkages changed in type, indicating fluid service systems. CONCLUSIONS: Linkages between community agencies serving homeless persons with comorbid psychiatric and substance use disorders are not extensive. However, they increased slightly under the first two years of the ACCESS program, and there are good reasons to anticipate greater improvements in the future. PMID- 9856619 TI - Costs and utilization of substance abuse care in a privately insured population under managed care. AB - OBJECTIVE: Cost and utilization patterns of substance abuse and mental health treatment under private, employer-sponsored, managed behavioral health care plans were examined. METHODS: Data were from claims made in 1995 in 93 behavioral health care plans covering 617,133 members. Rates of use of mental health and substance abuse care were determined, as were payments by insurers and patients for the two types of care. Means were calculated per plan member and per user of either of these service types. RESULTS: Approximately .3 percent of plan members used any substance abuse services; 5.2 percent used mental health services. However, among substance abuse patients, average costs were more than twice as high as average costs for mental health patients. For substance abuse treatment, the annual cost per user was $2,188, compared with $979 for users of mental health care. Annual per-member costs were $6.51 for substance abuse treatment and $50.08 for mental health care. Higher costs for substance abuse treatment reflected greater rates of use of both inpatient and intensive outpatient treatment. Overall, substance abuse costs represented 13 percent of insurance payments for behavioral health care and perhaps .4 percent of the cost of health insurance overall. CONCLUSIONS: Substance abuse coverage accounts for a small fraction of insurance payments for behavioral health coverage and a very small fraction of insurance payments for both physical and behavioral health care. PMID- 9856620 TI - Implementing publicly funded risk contracts with community mental health organizations. AB - The study analyzed the experience of the Los Angeles County Department of Mental Health with implementation of new contractual arrangements for services for patients with severe mental illness. The arrangements shifted the financial risk for treatment to community organizations and paid a fixed annual rate per enrolled patient without further adjustment for severity of illness. Patients were assigned to the program based on high prior treatment costs. The new contractual approach enhanced programs' flexibility and accountability and increased their emphasis on principles of psychosocial rehabilitation. Challenges in implementation included disenrollment of the majority of assigned patients by the community organizations at risk for high treatment costs. Prior treatment costs for continuing cases, while high, were lower than those for disenrolled cases. Existing information systems provided limited clinical and cost data, making it difficult to monitor providers' performance. Risk contracting required substantial clinical, fiscal, and management changes at community organizations and the mental health authority. The analysis suggests that mental health authorities that are planning to institute risk contracts need to balance fiscal incentives with performance guarantees and to pay particular attention to information systems requirements and to the severity of patients' illness. Although risk contracts present challenges, they can lead to improvements in service delivery that persist beyond the implementation phase. PMID- 9856621 TI - Intensity and duration of intensive case management services. AB - OBJECTIVE: Two policy issues related to intensive case management programs were examined: limiting caseload size to ensure that services are intensive and providing intensive services to the same clients in perpetuity. METHOD: The Denver Acuity Scale, which indicates need for services on a 5-point scale, was used for determining the service intensity needed by consumers and for increasing case managers' efficiency when caseload size varied. The acuity ratings of individuals in the Denver intensive case management program were examined to evaluate the effects of service duration on decompensation. Each consumer was rated at every service contact. The percentage of individuals readmitted to the program after they graduated was calculated, and the trajectories of 87 individuals who continued to be served by the program after they attained the highest rating of functioning were examined. RESULTS: Of the 112 individuals who graduated from the program in the 29-month study period, four (4 percent) were readmitted. More than half of the 87 consumers who achieved the highest functioning level did not deviate from that level for the remainder of the study period. Nearly a fifth showed some deterioration shortly after achieving that level but then improved. Slightly more than a fourth continued to deteriorate, but many never reached the lowest levels of functioning. CONCLUSIONS: To increase efficiency and ensure appropriate service levels, service intensity should be based on individual consumers' functioning levels. Most consumers are unlikely to need intensive case management in perpetuity. Providing more intensive services than needed or providing services longer than needed is inefficient and may even impede consumer recovery. PMID- 9856622 TI - Health status and health care costs for publicly funded patients with schizophrenia started on clozapine. AB - OBJECTIVE: The study examined the effect of clozapine treatment on the health care costs and health status of people with schizophrenia who are supported by public funds. METHODS: Thirty-three patients with schizophrenia hospitalized in a state facility were interviewed within one week of starting clozapine and six months later. Health status was assessed with four clinical rating scales measuring severity of psychopathology, negative symptoms, depression, and quality of life. Cost and health care utilization data were collected for the six months before and after initiation of clozapine. RESULTS: Only 52 percent of the subjects stayed on clozapine for six months. Subjects who continued on clozapine were more likely to be discharged within six months than those who did not continue. Six months after clozapine was started, health care costs showed a sayings of $11,464 per person, even after adjustment for pretreatment costs, and health status was improved. CONCLUSIONS: For subjects who continued on clozapine for six months, clozapine treatment was associated with reduced days of psychiatric hospital care, reduced overall costs despite increased outpatient treatment and residential costs, and improved health status. PMID- 9856623 TI - Psychiatrists' and internists' knowledge and attitudes about delivery of clinical preventive medical services. AB - OBJECTIVE: Changes in the health care environment have placed a greater responsibility on psychiatrists to deliver basic primary care services. The study assessed baseline knowledge and attitudes about clinical preventive medical services among psychiatric faculty and psychiatric residents at a tertiary care medical center. METHODS: Residents and faculty in psychiatry and general internal medicine completed a structured questionnaire, including 20 case scenarios, that assessed their baseline knowledge of clinical preventive medical services, their attitudes concerning delivery of those services, and their beliefs about the effectiveness of those services in changing patients' behavior. The case scenarios and knowledge questions were based on the clinical preventive medical services recommendations outlined by the U. S. Preventive Services Task Force. RESULTS: Psychiatrists reported more frequent assessment of and counseling about the use of illicit drugs and weapons, and internists were more likely to query about measures related to physical health such as cancer screening and immunizations. The two groups reported similar attitudes toward the need for and the efficacy of preventive medical services. Commonly cited barriers to the delivery of preventive care included lack of time and education. Psychiatrists scored reasonably well on baseline knowledge about guidelines for preventive medical services, particularly given their recent lack of specific education in these matters. CONCLUSIONS: Psychiatrists believe clinical preventive services are important and express interest in their delivery. Additional educational interventions are needed to train psychiatrists in clinical preventive services to avoid missed clinical opportunities for intervention in psychiatric populations that may have poor access to other medical care. PMID- 9856624 TI - A comparison of federal definitions of severe mental illness among children and adolescents in four communities. AB - OBJECTIVE: Using data from an epidemiological survey, the study compared existing definitions of severe mental illness and serious emotional disturbance among children and adolescents to demonstrate the range of prevalence rates resulting from application of different definitions to the same population. METHODS: Three definitions of severe mental illness and serious emotional disturbance were applied to data from the Methods for the Epidemiology of Child and Adolescent Mental Disorders survey, with a sample of 1,285, conducted in 1991-1992 by the National Institute of Mental Health. The resulting proportions of cases identified, demographic characteristics, service use, and perceived need for services were compared. RESULTS: From 3 to 23 percent of the sampled youth met criteria for severe mental illness or serious emotional disturbance. From 40 percent to as many as 78 percent of the defined youth used a mental health service in the year before the survey. School and ambulatory specialty settings were used most frequently. Generally, more than half of the parents of children with severe mental illness or serious emotional disturbance thought that their child needed services. CONCLUSIONS: The prevalence and characteristics of severe mental illness and serious emotional disturbance among children are sensitive to the definition used and its operationalization. Care should be taken by policy makers and service planners to avoid either over- or underestimating the prevalence of impaired youth in need of intensive interventions. PMID- 9856625 TI - Posttraumatic stress disorder and other predictors of health care consumption by Vietnam veterans. AB - A total of 641 randomly selected Australian veterans of the Vietnam War were interviewed about their use of health care in the previous two weeks to determine what factors contributed to health care consumption. Seventy-three variables were examined by univariate linear regression and then grouped into seven categories relating to age, physical and mental health, predisposition to posttraumatic stress disorder (PTSD), deployment and repatriation experiences, and membership in veterans groups. PTSD was associated with an additional cost of $79 in health care for the two-week period. Each physical diagnosis was associated with an additional $28. Alcohol consumption was not related to health care costs. Other important variables contributing to costs were depression, educational status, the quality of the repatriation experience, and social support. PMID- 9856626 TI - Psychosocial characteristics of pregnant and nonpregnant HIV-seropositive women. AB - Thirty-three HIV-positive women, 12 of whom were pregnant, participated in semistructured interviews to define areas of psychosocial need. Eighty-eight percent of the subjects reported current unemployment. A history of substance abuse was reported by 82 percent, suicide attempts by 52 percent, and sexual problems by 43 percent. Approximately 30 percent reported elevated levels of depressive symptoms on standardized symptom inventories. The pregnant women appeared psychologically healthier than the nonpregnant group. HIV-positive women face multiple psychosocial stressors and may experience significant psychological distress. PMID- 9856627 TI - Psychiatric hospitalization of persons with dual diagnoses: estimates from two national surveys. AB - Individuals with both mental illness and alcohol or drug use disorders present distinctive treatment and human service problems, including increased risk for psychiatric hospitalization in community hospitals. Using national hospital discharge abstract data for 1990 and 1994, this study compared differences in psychiatric hospitalization in community hospitals of patients with mental illness only and those with mental disorders and substance use disorders. Individuals with dual diagnoses were younger, and a greater proportion were men. Medicaid was the primary payer for a larger percentage of those with dual diagnoses. Nationally, the number of community hospitalizations for dually diagnosed patients increased 15 percent from 1990 to 1994, and total hospital charges increased from $1.9 to $2.2 billion. PMID- 9856628 TI - Two cases of suicide attempt by patients with Down's syndrome. AB - This report documents cases of suicide attempts by two persons with Down's syndrome. The patients experienced dysphoric affect and feelings of hopelessness about unmet needs associated with their disability. During the course of a major depressive episode, each individual made a suicide attempt that could have been fatal. Although suicidal ideation and attempts are infrequent among patients with mental retardation, completed suicides and potentially fatal attempts have been reported. It is vitally important that mental health practitioners appreciate the seriousness of suicidal ideation among persons who have mental retardation and treat the underlying psychiatric disorder. PMID- 9856629 TI - More on profit and quality in managed care. PMID- 9856630 TI - Use of the BASIS-32. PMID- 9856631 TI - Consumer satisfaction. PMID- 9856633 TI - Five-state review of CMHC partial hospital programs finds $229 million in questionable medicare payments. PMID- 9856632 TI - Overoptimism about drugs. PMID- 9856634 TI - Mental health, substance abuse programs receive 15 percent funding hike. PMID- 9856635 TI - Guide helps public-sector agencies draft stronger managed care contracts. PMID- 9856636 TI - NMHA, NAMI launch separate initiatives to improve community care for adults with severe mental illness. National Mental Health Association. National Alliance for the Mentally Ill. PMID- 9856637 TI - Tracking the epidemiology of antimicrobial drug resistance in hospitals: time to deploy molecular typing. PMID- 9856638 TI - A spectrum of verocytotoxigenic Escherichia coli. PMID- 9856639 TI - New perspectives on the role of Escherichia coli O157:H7 and other enterohaemorrhagic E. coli serotypes in human disease. AB - This review compares the rates of detection of non-O157:H7 enterohaemorrhagic Escherichia coli (EHEC) with EHEC O157:H7 in outbreaks and sporadic cases of human disease by analysing Australian data and the world literature. Numerous outbreaks of disease have been attributed to EHEC O157:H7. In many studies, isolation rates of this organism have been low and attempts to seek other EHEC have not been made. Ease of isolation and identification of the O157:H7 serotype may have given the impression that this serotype was the sole organism responsible for the outbreaks. Careful review and analysis shows that serotypes other than O157:H7 also play an important role in human disease. Evidence is presented from several overseas outbreaks described in the literature, as well as from investigations of the Adelaide O111:H- outbreak, that suggests an association between severity of disease and multiple infecting serotypes. While not diminishing the role of the O157:H7/H- clone, this review indicates that other serotypes can be responsible for outbreaks as well as cases of sporadic human disease. The current focus on O157:H7 has major implications in terms of diagnosis, the food industry and human health. PMID- 9856640 TI - The relative pathogenicity of Candida krusei and C. albicans in the rat oral mucosa. AB - The relative pathogenicity of Candida krusei and C. albicans was investigated by assessing their colonisation and infectivity of the Sprague-Dawley rat oral mucosa. During an initial 21-week period with intermittent oral inoculation, both Candida spp. demonstrated variable surface colonisation of the oral mucosa. After 3 days of oral inoculation, both yeast species were recovered from all animals. During the 21-week period the mean oral load of C. albicans in the control group of rats varied between (26-274) x 10(1) cfu/ml whereas the two test groups of rats carrying C. krusei CK9 and CK13 had a mean load of (2-10) x 10(1) cfu/ml. Although oral colonisation by C. albicans was greater than that of C. krusei, neither species induced candidal infection during this period. Subsequent immunosuppression of the rats by intramuscular cyclophosphamide (40 mg/kg body weight) initiated C. albicans infection of the dorsal tongue (around the conical papillae area) after 4 weeks, in all of three animals while similar lesions due to C. krusei were seen--albeit after 5-7 weeks--in three of eight animals. Characteristic histological changes of mucosal candidosis were discernible on the lingual mucosa of rats infected with both Candida spp. including parakeratosis, absence of a stratum granulosum, thickened rete ridges, micro-abscess formation and polymorph infiltration of the lingual epithelium. Although both species produced fungal hyphae that penetrated the epithelium up to the prickle cell layer, C. albicans hyphae tended to be relatively more profuse. Taken together these results substantiate, for the first time in an animal model, the clinical evidence that C. krusei, once considered an innocuous commensal, is capable of transforming into an invasive pathogen under conditions of immunosuppression. PMID- 9856641 TI - Secondary structure and molecular analysis of interstrain variability in the P5 outer-membrane protein of non-typable Haemophilus influenzae isolated from diverse anatomical sites. AB - The sequence of the non-typable Haemophilus influenzae (NTHi) P5 outer-membrane protein from a range of clinical isolates is presented and represents the first analysis of the heterogeneity in P5 from NTHi isolates from diverse anatomical sites. The basis of the previously observed inter-strain variation in the electrophoretic mobility is attributed to heterogeneity in three hypervariable regions. Alignment of the P5 sequences identified regions which are highly conserved and align with the transmembrane region predicted for the homologous Escherichia coli protein, OmpA. Variable regions correspond to surface-exposed loops, of which the first loop falls into subclasses. However, these subclasses fail to correlate with anatomical predisposition. Although P5 has been proposed as a fimbrial protein composed of coiled coils, both structural analysis by circular dichroism of purified P5 and computer analysis of the multiply aligned sequences predict a high proportion of beta strand with no evidence of coiled coil structure. A detailed model of P5 is presented. PMID- 9856642 TI - Ultrastructure of Calymmatobacterium granulomatis: comparison of culture with tissue biopsy specimens. AB - The ultrastructural features of cells of Calymmatobacterium granulomatis from monocyte co-cultures and tissue biopsy specimens were compared. In cultures the bacteria were mainly extracellular, i.e., not within membrane-bound vacuoles. The bacterial body was surrounded by a uniformly extensive homogeneous layer with a relatively high electron density. This layer varied considerably in tissue biopsy specimens, having either homogeneously electron-dense or delicate web-like structures with varying density and thickness. In tissue specimens the bacteria were located predominantly within vacuoles of varying sizes in the cytoplasm of the macrophages and, occasionally, extracellularly within the intercellular spaces of the stroma. The bacterial cytoplasm contained ribosomes scattered throughout with electron-dense granules located peripherally. The trilaminar cell wall structure was typical of a gram-negative organism, comprising an outer membrane, a middle electron-opaque layer and an inner plasma membrane. Surface structures such as fimbriae, flagella and bacteriophages were not identified in specimens from either source. PMID- 9856643 TI - Microbiology of liver and spleen abscesses. AB - To study the aerobic and anaerobic microbiology of liver and spleen abscesses and correlate the results with predisposing factors, potential causes and routes of infection, clinical and laboratory data of 48 patients with liver abscesses and 29 with spleen abscesses treated between 1970 and 1990 were reviewed retrospectively. In liver abscesses, a total of 116 isolates (2.4 isolates/specimen) was obtained; 43 were aerobic and facultative species (0.9 isolates/specimen) and 73 were anaerobic species or microaerophilic streptococci (1.5 isolates/specimen). Aerobic bacteria only were isolated from 12 (25%) abscesses, anaerobic bacteria only from eight (17%), and mixed aerobic and anaerobic bacteria from 28 (58%); polymicrobial infection was present in 38 (79%). The predominant aerobic and facultative isolates were Escherichia coli (11 isolates), Streptococcus group D (8), Klebsiella pneumoniae (5) and Staphylococcus aureus (4). The predominant anaerobes were Peptostreptococcus spp. (18 isolates), Bacteroides spp. (13), Fusobacterium spp. (10), Clostridium spp. (10) and Prevotella spp. (4). There were 12 isolates of micro-aerophilic streptococci. S. aureus and beta-haemolytic streptococci were associated with trauma; Streptococcus group D, K. pneumoniae and Clostridium spp. with biliary disease; and Bacteroides spp. and Clostridium spp. with colonic disease. In splenic abscesses, a total of 56 isolates (1.9 isolates/specimen) was obtained; 23 were aerobic and facultative species (0.8 isolates/specimen), 31 were anaerobic species or micro-aerophilic streptococci (1.1 isolates/specimen) and two were Candida albicans. Aerobic bacteria only were isolated from nine (31%) abscesses, anaerobic bacteria from eight (28%), mixed aerobic and anaerobic bacteria from 10 (34%) and C. albicans in two (7%); polymicrobial infection was present in 16 (55%). The predominant aerobic and facultative isolates were E. coli (5 isolates), Proteus mirabilis (3), Streptococcus group D (3), K. pneumoniae (3) and S. aureus (4). The predominant anaerobes were Peptostreptococcus spp. (11 isolates), Bacteroides spp. (5), Fusobacterium spp. (3) and Clostridium spp. (3). S. aureus, K. pneumoniae and Streptococcus group D were associated with endocarditis, E. coli with urinary tract and abdominal infection, Bacteroides spp. and Clostridium spp. with abdominal infection and Fusobacterium spp. with respiratory infection. PMID- 9856644 TI - Interference in initial adhesion of uropathogenic bacteria and yeasts to silicone rubber by a Lactobacillus acidophilus biosurfactant. AB - The ability of the Lactobacillus acidophilus RC14 biosurfactant 'surlactin' to inhibit the initial adhesion of various uropathogenic bacteria and two yeast strains to silicone rubber was investigated in a parallel-plate flow chamber in filter-sterilised pooled human urine. A parallel-plate flow chamber with a silicone rubber bottom plate was filled with a 1.0 mg/ml biosurfactant solution for adsorption overnight (18 h). Subsequently, the adhesion of the bacterial or yeast cells from a urine suspension under low flow (shear rate 15 s(-1)) was followed in situ by automated image analysis. Control tests were with untreated silicone rubber. Initial deposition rates and numbers of adhering cells after 4 h of flow were determined. Surlactin layers caused a marked inhibition of the initial deposition rates and adhesion numbers after 4 h for the majority of the bacteria (11 of 15 strains tested) and this inhibition was particularly effective against Enterococcus faecalis, Escherichia coli and Staphylococcus epidermidis. Although the initial deposition rates of the two Candida albicans strains were reduced by c. 50% in comparison with the controls, the numbers of yeast cells adhering after 4 h were similar. PMID- 9856645 TI - Purification of immunosuppressive factor from Capnocytophaga ochracea. AB - Capnocytophaga, one of the genera of oral bacteria, has been implicated in the pathogenesis of several diseases, including endocarditis, septicaemia and disorders of the oral cavity such as abscesses and periodontal disease. This study examined sonic extracts (SE) of Capnocytophaga strains for their ability to alter lymphocyte function. The SE of tested Capnocytophaga caused dose-dependent suppression of spleen cells in response to mitogen. This suppressive effect was heat-labile and sensitive to the proteolytic enzyme pronase E. The suppressive factor (SF) was purified from SE of C. ochrasea by a combination of ultrogel AcA34, high-pressure liquid DEAE ion-exchange chromatography and hydroxyapatite columns, which revealed a single band of 14 kDa by SDS-PAGE. Rabbit anti-serum against the purified SF inhibited the immunosuppression induced by SE of C. ochracea with the recovery of lymphocyte proliferation. PMID- 9856646 TI - Different O and K serotype distributions among clinical and environmental strains of Serratia marcescens. AB - Recent revision of the O serotyping scheme for Serratia marcescens has allowed the definitive serological identification of a collection of 511 epidemiologically distinct strains in terms of both lipopolysaccharide (O) antigens and capsular (K) antigens. High levels of typability were achieved, 88% and 91% respectively, with only 2% failing to type with either method. In most cases, non-typability was due to a lack of antigen, i.e., the strains produced only rough LPS or were acapsular, suggesting that typability would be little improved by the discovery of additional serotypes. The distribution of the 58 O:K serotypes was very uneven, with O14:K14 accounting for 30% of the 423 clinical strains in the collection, but only 5% of the 88 non-clinical, environmental strains. Thus, the prevalence of O14:K14 strains in hospitals is not reflected in the environment. Similar conclusions were valid for O27:K14, O21:K3 and O21:K14 strains, as well as those with rough lipopolysaccharide. Conversely, the proportions of O6:K3, O6:K14, O8:K14 or O28:K28 strains were significantly lower among the clinical collection than among their environmental counterparts (12% in total rather than 65%). This suggests that O14:K14 may have a selective advantage in colonising or infecting hospitalised patients and, therefore, that the O14 and K14 polysaccharides themselves may contribute towards the apparent pathogenicity of these serotypes. PMID- 9856647 TI - Antibiotic resistance and putative virulence factors of Serratia marcescens with respect to O and K serotypes. AB - Serratia marcescens serotypes O6:K14, O8:K14 and O28:K28 are common in the natural environment, but rare in hospitals. Serotypes O14:K14 and O27:K14 predominate among clinical strains, but not in the environment, suggesting that the latter serotypes may be more suited for survival in the clinical setting. Consequently, 469 epidemiologically distinct strains of S. marcescens were tested for various putative virulence factors and analysed for associations with serotype. The factors positively associated with serotype O14:K14 were agglutination of five different species of red blood cells and expression of type 1 fimbriae. These were found in 63% and 53% of O14:K14 strains, respectively, compared with 7% and 12% of the three 'environmental serotypes'. Almost a quarter of the collection expressed the mannose-resistant haemagglutinin indicative of type 3 fimbriae, but this was not associated with any serotype. The production of DNAase, haemolysin, lipase, lecithinase, proteases and siderophores was almost universal and showed no serotype correlations. Almost half of the strains (46%) were resistant to serum and serotypes O27:K14 and O6:K14 were strongly associated with this characteristic. Serotype O27:K14 was also associated with higher proportions of antibiotic-resistant strains than other serotypes, but the same was not true of serotype O14:K14. All three 'environmental serotypes' were associated with low frequencies of antibiotic resistance; <12% were resistant to gentamicin, carbenicillin or piperacillin, or any combination of these three, compared with 20-25% of O14:K14 strains and >42-51% of O27:K14 strains. Pigment production was strongly associated with serotype. None of the O14:K14 or O27:K14 strains produced prodigiosin, but frequencies for the three 'environmental serotypes' ranged from 31% of O28:K28 strains to 85% of O6:K14 strains. The results of this study suggest that the adherence capability of S. marcescens strains may play a role in the colonisation of hospital patients, while the production of prodigiosin is a marker of environmental origin. PMID- 9856648 TI - Comparative susceptibilities of various AIDS-associated and human urogenital tract mycoplasmas and strains of Mycoplasma pneumoniae to 10 classes of antimicrobial agent in vitro. AB - The susceptibilities of 40 strains of various Mycoplasma species to 10 classes of antimicrobial agents were compared in vitro by a broth microdilution method. The strains tested comprised 20 strains of four AIDS-associated species--M. penetrans (1 strain), M. fermentas (5 strains), M. pirum (6 strains) and M. genitalium (8 strains)--nine strains of the urogenital tract species M. hominis and 11 strains of M. pneumoniae. The results demonstrated wide variation in the susceptibilities of the different Mycoplasma spp. to different classes of antimicrobial agent. All the mycoplasmas were susceptible or highly susceptible to the fluoroquinolones, with sparfloxacin the most active, and to the diterpine antibiotic tiamulin. M. pneumoniae and M. genitalium strains were also highly susceptible to the macrolides, particularly azithromycin and had similar antibiotic susceptibility patterns to most other antimicrobial agents. However, all strains of M. genitalium were resistant to streptomycin (MIC 250->500 mg/L) whereas all M. pneumoniae isolates, except the MAC strain, were susceptible (MICs 1.25-12.5 mg/L). M. pirum isolates varied considerably in their susceptibility to macrolides (MIC range versus azithromycin 0.0025->100 mg/L). M. fermentans strains were susceptible to the tetracyclines, lincosamides and mupirocin, but varied in susceptibility to aminoglycosides. Most M. hominis strains were susceptible to the tetracyclines and all were susceptible to clindamycin and mupirocin. M. penetrans GTU 54 was susceptible to azithromycin, the tetracyclines and lincosamides as well as to the fluoroquinolones and tiamulin. PMID- 9856649 TI - Severe gastritis in guinea-pigs infected with Helicobacter pylori. AB - An appropriate animal model is essential to study Helicobacter pylori infection. The aim of this study was to investigate if H. pylori can colonise the guinea-pig stomach and whether the infection causes gastritis and a serological response similar to that observed in man. Guinea-pigs were infected either with fresh H. pylori isolates from human gastric biopsies or with a guinea-pig passaged strain. When the animals were killed, 3 and 7 weeks after inoculation, samples were taken for culture, histopathology and serology. H. pylori was cultured from 22 of 29 challenged animals. All culture-positive animals exhibited a specific immune response against H. pylori antigens in Western blotting and gastritis in histopathological examination. Antibody titres in enzyme immunoassay were elevated among animals challenged with H. pylori. The inflammatory response was graded as severe in most animals and consisted of both polymorphonuclear leucocytes and lymphocytes. Erosion of the gastric epithelium was found in infected animals. These results suggest that the guinea-pig is suitable for studying H. pylori-associated diseases. Moreover, guinea-pigs are probably more similar to man than any other small laboratory animal as regards gastric anatomy and physiology. PMID- 9856650 TI - Effect of inhibitors in clinical specimens on Taq and Tth DNA polymerase-based PCR amplification of influenza A virus. AB - Fifteen randomly selected nasopharyngeal (NP) swab specimens (culture-negative for influenza A virus) were spiked with influenza A virus and the nucleic acids were extracted and subjected to PCR amplification with Thermus aquaticus (Taq) and T. thermophilus (Tth) DNA polymerases. Products of the expected size, and giving equivalent band intensities, were obtained from four specimens with both polymerases. Fox six specimens, less products were obtained with Taq DNA polymerase than with Tth DNA polymerase. Products were detected from five NPs only by PCR with Tth DNA polymerase. The transport medium and the calcium alginate swab fibre of the specimens were shown not to be the source of the inhibitors. The incorporation of 32P-dCTP into cDNA, and the yield of PCR products of cDNA made from control RNA template (purified from H2O spiked virus suspension) were decreased in the presence of inhibitory extracts, showing that both the reverse transcription (RT) and PCR steps in amplification with Taq DNA polymerase were sensitive to the inhibitors. In contrast, Tth DNA polymerase was more resistant to the inhibitors and viral nucleic acid from all the specimens examined could be amplified and detected in a single step by RT-PCR with Tth DNA polymerase. PMID- 9856651 TI - Phase I-II study of 5-fluorouracil, leucovorin, doxorubicin, methotrexate, and long-term oral etoposide (FLAME) in unresectable or metastatic gastric cancer. AB - The objective of this phase I-II study was to determine the efficacy and toxicity of combination chemotherapy with 5-fluorouracil, leucovorin, doxorubicin, methotrexate, and oral etoposide (FLAME) in patients with measurable unresectable or metastatic gastric cancer. Starting doses on the phase I study were as follows: methotrexate 50 mg/m2 intravenous bolus day 1; leucovorin 20 mg/m2 intravenous bolus days 2 through 4, starting 24 hours after the methotrexate dose; 5-fluorouracil 325 mg/m2 intravenous bolus 15 minutes after leucovorin days 2 through 4; doxorubicin 25 mg/m2 intravenous bolus day 8; and oral etoposide 50 mg/day for 14 days, starting on day 8. A new cycle started on day 28. A total of 42 patients were treated--10 patients in the phase I study and 32 patients in the phase II study. Dose-limiting toxicity was encountered in the phase I study on the second escalation step, when doxorubicin was escalated to 30 mg/m2 and 5 fluorouracil was escalated to 350 mg/m2. In the phase II study 28 patients (109 courses) were evaluable for toxicity. Neutropenia grade 3 or more was dose limiting and was documented in 12 patients (43%) during 22 treatment courses (20%). Neutropenia was associated with febrile neutropenia requiring hospitalization in four patients during five courses of therapy. Grade 3 stomatitis and grade 3 diarrhea was infrequent, documented in two patients (two courses) and three patients (four courses), respectively. All other toxicity was grade 1 and grade 2. The combined objective response rate in 38 evaluable patients entered in both studies was 23.3% (six partial responses and one complete response). Stable disease was documented in 15 patients (39.5%). The median survival for the 42 patients entered in both trials was 6.9 months (95% confidence interval, 5.9-8.5 months). The objective response rate and median survival for the combined group is comparable with that reported for the etoposide, leucovorin, and 5-fluorouracil (ELF), and 5-fluorouracil and methotrexate (FMTX) regimens in a recently reported, multicenter, phase III study. PMID- 9856652 TI - The addition of paclitaxel to continuous infusion 5-fluorouracil is an active regimen for metastatic breast cancer. AB - Metastatic breast cancer is commonly thought to be incurable. Treatment advances have resulted in increased response rates, although such responses are often more palliative than curative. A regimen of continuous infusion 5-fluorouracil (5FU) or continuous infusion 5-fluorouracil with paclitaxel was studied in patients with metastatic breast cancer and measurable disease. The induction therapy preceded high-dose ifosfamide, carboplatin, and melphalan in a phase I-II trial. Eighty-seven patients were enrolled in the trial. Forty-five received continuous infusion 5-fluorouracil as induction and 42 received 5-fluorouracil and paclitaxel. The single-agent, continuous infusion 5-fluorouracil cohort had one complete response (2%) and eight partial responses (18%). The combination continuous infusion 5-fluorouracil and 3-hour paclitaxel regimen produced four complete responses (10%) and 17 partial responses (40%). The combination regimen of continuous infusion 5-fluorouracil with bolus paclitaxel was well tolerated and with a 50% response rate, is an active regimen for women with metastatic breast cancer. PMID- 9856653 TI - A phase II trial and pharmacokinetic analysis of 96-hour infusional paclitaxel in patients with metastatic colorectal cancer. AB - A phase II study was conducted to evaluate the activity and toxicity of 96-hour infusional paclitaxel in patients with previously untreated metastatic colorectal cancer. Twelve patients were enrolled in this study. The first patient received a total dose of 140 mg/m2 over 96 hours resulting in grade 4 neutropenia, neutropenic fever, and grade 3 stomatitis. Subsequent patients received a total dose of 120 mg/m2 over 96 hours. Grade 3 to 4 neutropenia occurred in four of these patients. No significant thrombocytopenia was observed. Grade 3 to 4 nonhematologic toxicities in the group treated at 120 mg/m2 over 96 hours included nausea/vomiting in one patient, stomatitis in one patient, and diarrhea in two patients. One patient experienced a possible pulmonary hypersensitivity reaction. None of the 12 patients achieved an objective response. Two patients had stable disease and ten had progressive disease. Pharmacokinetic parameters including maximum plasma concentration and area under the concentration time curve were significantly higher in patients with grade 3 to 4 neutropenia than patients who experienced less toxicity. The authors conclude that further study of 96-hour infusional paclitaxel in patients with metastatic colorectal carcinoma is not warranted. PMID- 9856654 TI - Paclitaxel as the initial treatment of multiple myeloma: an Eastern Cooperative Oncology Group Study (E1A93). AB - The authors assess the activity and toxicity of paclitaxel in previously untreated patients with multiple myeloma. Eighteen patients with previously untreated multiple myeloma were enrolled. Paclitaxel was given in a dose of 250 mg/m2 by a continuous intravenous infusion for 24 hours every 21 days for four cycles. All patients received granulocyte colony stimulating factor in a dose of 5 microg/kg each day until the absolute neutrophil count was 10,000/mm3. All patients were evaluated after four cycles. Four (29%) objective responses were observed in the 14 eligible patients. No complete responses occurred. Three lethal toxicities were observed, two were the result of neutropenic sepsis. Sixty one percent of patients experienced some type of severe nonhematologic toxicity. Patients who received four cycles of paclitaxel were given further treatment at the discretion of the investigator. The median survival of all eligible patients was 2.8 years, which is comparable with the median survival with melphalan and prednisone of 2.3 years or vincristine, carmustine, melphalan, cylophosphamide, and prednisone of 2.4 years. Paclitaxel in the dosage used in this study has prohibitive toxicity. The four (29%) responses in 14 evaluable untreated patients indicates that paclitaxel is active in the treatment of multiple myeloma. No complete remissions were recorded. Further studies using paclitaxel in a smaller dose, in combination with other agents, or using one of the paclitaxel analogs may be useful in the treatment of multiple myeloma. PMID- 9856655 TI - Gangliogliomas: experience with 34 patients and review of the literature. AB - Ganglioglioma is an uncommon central nervous system tumor. The role of adjuvant postoperative radiation therapy is undefined. The authors retrospectively reviewed the clinicopathologic features and results of therapy for 34 patients with ganglioglioma treated at the University of California at Los Angeles. There were 18 women and 16 men. Median age was 18 years. Twenty-five tumors were low grade. Twenty-one patients underwent gross total resection. Three patients received adjuvant radiotherapy. The 4-year actuarial progression free and overall survival rates were 67% and 75%, respectively. The median time to progression was 14 months and all relapses were local. Factors significantly influencing progression-free or overall survival according to univariate analysis included degree of resection and tumor grade. Survival and relapse were not significantly influenced by any factor according to multivariate analysis. The progression-free survivals after gross total resection of low- and high-grade tumors were 78% and 75%, respectively. Respective rates after subtotal resection were 63% and 25%. Review of the literature demonstrates no role for radiotherapy after total resection of ganglioglioma or after partial removal of low-grade tumor. Radiation therapy appears to reduce the relapse rate after partial removal of high-grade lesions. A dose in excess of 5,000 cGy is necessary for ganglioglioma. PMID- 9856656 TI - High-dose megestrol acetate in the treatment of patients with ovarian cancer who have undergone previous treatment: Eastern Cooperative Oncology Group Study PD884. AB - High-dose megestrol acetate has been reported to be effective salvage therapy for women with ovarian carcinoma. The Eastern Cooperative Oncology Group performed this phase II study of oral megestrol acetate, 200 mg four times daily until disease progression, in 33 patients either with stage III or IV histologically confirmed ovarian carcinoma or with unresectable tumor in the pelvis with measurable or evaluable disease who progressed after treatment with one prior chemotherapy regimen. Thirty and 31 patients were evaluable for response and toxicity, respectively. No patient had an objective response and none had subjective improvement after a median treatment period of 1.4 months. Nausea or vomiting occurred in most patients, usually grade 1-2. Megestrol acetate is ineffective salvage therapy for patients with inoperable, previously treated ovarian carcinoma. PMID- 9856657 TI - Metastatic melanoma from intraocular primary tumors: the Southwest Oncology Group experience in phase II advanced melanoma clinical trials. AB - Ocular melanoma is an uncommon malignancy that, in the presence of metastatic disease, has a poor prognosis for response to treatment and survival. Patients with ocular melanoma are often excluded from clinical trials because of the impression that these patients have a poorer response rate to treatment with anticancer agents and poorer survival, possibly related to the predominance of the liver as a site of metastasis. Sixty-four eligible patients with advanced melanoma arising from ocular primary tumors were entered into seven phase II clinical trials of anticancer therapy activated by the Southwest Oncology Group (SWOG) during the 1980s. Eligible patients with nonocular primaries entered into these trials (420 patients) served as a comparison group for survival, pretreatment characteristics, and response rates. Multivariate Cox model analysis of survival data (with survival from the time of study registration as the primary end-point) was conducted. Among the 484 patients observed, patients with ocular melanoma were older than those with nonocular primary tumors and were more likely to have visceral metastasis, metastasis to the liver, and only one metastatic site at registration, primarily to viscera and liver. The median overall survival after registration to study for both groups was 5 months. There was no significant difference in overall survival between patients with ocular melanoma and those with nonocular melanoma after adjusting for a number of prognostic factor (p = 0.43). Furthermore, the overall objective response rate of patients with ocular melanoma in these studies was not significantly different from that achieved in the nonocular group (9% vs. 11%; p = 1.00). Patients with advanced ocular or nonocular melanoma have similar response rates and survival in this series of cooperative group phase II trials. Patients with ocular primaries should not be excluded from investigational studies in advanced melanoma. PMID- 9856658 TI - Phase II study assessing the feasibility of using elemental supplements to reduce acute enteritis in patients receiving radical pelvic radiotherapy. AB - From June 1994 through September 1995, 17 patients with gynecologic cancer were entered into a feasibility study using elemental supplements (ES) during the course of pelvic radiotherapy. The level of compliance with the ES regimen was evaluated by assessing sachet counts and patients' weekly diaries. Diaries were also used to assess compliance with a modified diet program. Bowel function in 45 patients receiving radiotherapy outside this study was assessed concurrently. All patients were reviewed weekly to record bowel function, which was also recorded 1 year after radiotherapy. After treatment of all patients the grade, duration of enteritis, need for antidiarrheal agents, and the likelihood of resolution at 1 year after radiotherapy were analyzed. The study used compliance with the ES regimen as its primary endpoint. Compliance with the ES regimen was achieved in 76.5% of patients, and compliance with the modified diet was achieved regardless of whether the ES was tolerated. Patients not complying with the ES, or who did not receive ES, had higher grade diarrhea, which was longer in duration and was less likely to have resolved than patients in the compliant group. Elemental supplements are well tolerated when taken during a course of pelvic radiotherapy and cause a mild diarrhea, which resolves after treatment has been discontinued. The lack of enteritis 1 year later in patients who complied with ES suggests that reducing the grade of acute enteritis might influence the appearance of later bowel effects. PMID- 9856659 TI - A phase I-II trial of lovastatin for anaplastic astrocytoma and glioblastoma multiforme. AB - Malignant gliomas are thought to be highly dependent on the mevalonate pathway for cell growth. Lovastatin, a cholesterol-lowering drug, inhibits not only the rate-limiting step in the mevalonate pathway (hepatic hydroxymethyl glutaryl coenzyme A reductase), but also the prenylation of several key regulatory proteins including ras and the small guanosine triphosphate binding proteins. Therefore, from August 1994 through March 1996, 18 patients with either anaplastic glioma or glioblastoma multiforme were entered into a trial testing the safety of high-dose lovastatin with or without radiation. Although the response data is too premature to evaluate activity, the fact that high doses of lovastatin are well tolerated with concurrent radiation suggests that central nervous system toxicity will not be a significant limiting toxicity as more selective farnesyltransferase inhibitors are brought into the clinic as radiation sensitizers. PMID- 9856660 TI - Treatment results of unresected or partially resected desmoid tumors. AB - The authors compare the progression-free survival of patients with unresected or partially resected desmoid tumors treated with radiotherapy with those not given radiotherapy. A retrospective analysis and pathologic review was performed on 14 patients treated at the University of California Los Angeles School of Medicine from 1965 through 1992. Median follow-up was 6 years. The 6-year progression-free survival for irradiated patients was 100%, compared with 50% for those not irradiated (p = 0.04). Of the seven patients irradiated, only two had a complete response and one had a partial response. There was no difference in disease specific survival between patients irradiated and those not irradiated, because only 1 of 14 patients died of desmoid tumor progression, which caused airway obstruction. This data suggest that radiotherapy may improve the progression-free survival of patients with unresected or partially resected desmoid tumor; however, the number of patients in this series is small. Most patients did not have a complete response to radiotherapy. For patients with tumor adjacent to the airway in the neck or upper thorax, the authors recommend radiotherapy because of the potential for mortality. Otherwise, because tumor progression rarely causes death, one must consider whether the morbidity of treatment would outweigh the morbidity of disease progression. PMID- 9856661 TI - A phase II trial of isotretinoin and alpha interferon in patients with recurrent squamous cell carcinoma of the cervix: a Gynecologic Oncology Group study. AB - From January 1993 through January 1996, 37 patients with unresectable squamous carcinoma of the cervix were entered on study and scheduled to receive oral isotretinoin 1 mg/kg per day with subcutaneous alpha interferon 6,000,000 units/day. A course was defined as 4 continuous weeks of therapy. The mean number of four-course cycles delivered was 1.8. One patient was ineligible because of wrong cell type and two were never treated. Thus, 34 patients were evaluable for toxicity. Eight patients were inevaluable for response. Five did not receive a complete 4-week course and three did not have additional tumor measurements; thus 26 were evaluable for response. Prior radiotherapy had been given to 25 patients and prior chemotherapy to 23 patients. There was no grade 4 neutropenia. The incidence of Gynecologic Oncology Group (GOG) grade 3 granulocytopenia and thrombocytopenia was 8.8% and 5.8%, respectively. Six patients (17.6%) developed grade 3 or worse nausea and vomiting. Four (11.7%) patients developed grade 3 neurologic symptoms. There were no complete responses and one partial response. The overall response rate was 3.8% (95% confidence interval, 0.1-19.6%). In this pretreated population, isotretinoin and alpha interferon in the dose and schedule employed exhibit minimal activity. PMID- 9856662 TI - Fractionated carboplatin radiosensitization: a phase I dose-escalation study. AB - Carboplatin is one of the most common drugs used for radiochemotherapy of cancer. However, the best way to combine the drug with fractionated radiotherapy has not been established. In the present study the authors investigated which maximum tolerated daily bolus dose of carboplatin would allow safe radiopotentiation for 10 consecutive radiotherapy days, the scheme being repeated twice during the 6 weeks that a conventional radiotherapy scheme lasts. Seventy-two patients with lung or pelvis malignancies were included in a dose escalation study. Twenty-four patients comprised the first baseline cohort and were treated with radiotherapy alone. The daily dose of carboplatin was escalated starting from 38 mg/m2 daily (for 10 days) and increasing by 7 mg/m2 per day. Six patients were to be included in each cohort. All 12 patients treated at the 38 mg/m2 and 45 mg/m2 dose level completed two cycles of 10-day carboplatin treatment with no grade III-IV toxicity. Granulocyte colony-stimulating factor effectively averted the incidence of neutropenia and allowed the administration of the second carboplatin 10-day cycle in five of six patients at the 52 mg/m2 daily dose level. Platelet grade III-IV toxicity was observed in all 12 patients (six supported with granulocyte colony-stimulating factor and six with granulocyte colony-stimulating factor and recombinant human erythropoietin) treated at the 59 mg/m2 daily dose level and none of them received the second cycle of chemotherapy. Twelve patients were treated at the same dose level using amifostine 500 mg before carboplatin infusion. Two patients interrupted chemotherapy because of severe nausea and vomiting. Nine of 10 who accomplished the 10-day treatment had platelet levels more than 90,000/microl on day 28 and completed the second 10-day cycle without severe toxicity. Acute radiation toxicity did not increase in the carboplatin cohorts. In this study the authors established a high-dose fractionated carboplatin schedule that can be safely administered during radical radiotherapy. PMID- 9856663 TI - Strontium-89 (89Sr) analgesia for rare thymic carcinoid tumor with bony metastases. AB - The authors report the cases of two patients in whom strontium-89 (89Sr) was used to relieve diffuse metastatic bone pain. The type of cancer involved, thymic carcinoid tumor, is itself rare and the risk of its metastasizing to the bone is very low. Both patients showed a measure of response to treatment, suggesting that this analgesic method has value for some patients. The marked benefit of one patient for a total of 9 months was attributable to two 89Sr injections, whereas the other patient improved for only 5 weeks after one injection. PMID- 9856664 TI - Overwhelming 5-fluorouracil toxicity in patients whose diabetes is poorly controlled. AB - The author presents seven patients with diabetes and cites from the literature seven patients with diabetes with probable enhanced 5-fluorouracil toxicity. In the author's seven patients, the amount of toxicity was directly related to the degree of hyperglycemia. In three of these patients who had the most severe hyperglycemia, toxicity was overwhelming, and led to death in two patients. In the 1960s, researchers showed, in a series of in vitro experiments, that glucose increased the cytotoxicity of 5-fluorouracil. The author concludes that patients whose diabetes is poorly controlled are at increased risk for severe 5 fluorouracil toxicity. New labeling instructions should advise caution in using 5 fluorouracil in patients with diabetes. PMID- 9856665 TI - Angiosarcoma of the breast mimicking radiation dermatitis arising after lumpectomy and breast irradiation: a case report. AB - Angiosarcoma arising in the breast is a very rare malignancy. Of the small number of reported cases, most arose in the breast after breast irradiation. A case is reported of angiosarcoma arising in the breast more than 7 years after lumpectomy and breast irradiation for carcinoma of the breast. The initial appearance was very similar to the changes of late radiation dermatitis and the true nature of the malignant lesion was not known for 23 months. PMID- 9856666 TI - Phase II trial of topotecan for the treatment of mesothelioma. AB - The North Central Cancer Treatment Group designed a phase II trial to assess the efficacy and toxicity of topotecan in patients with unresectable malignant pleural mesothelioma. Twenty-two previously untreated patients with unresectable pleural mesothelioma and good performance status (Eastern Cooperative Oncology Group performance status 0, 1, or 2) were enrolled on this trial from October 1993 through July 1994. Nineteen men and three women, median age 66 years (range, 44-78 years), were treated with topotecan 1.5 mg/m2 intravenously over 30 minutes daily for 5 days at 3-week intervals until toxicity, progression of disease, or a patient decided to discontinue treatment. There were seven patients with measurable disease and 15 with evaluable disease; all were assessable for response and toxicity. A total of 113 cycles of treatment were given, for a median of three cycles (range, 1-26 cycles). Myelosuppression was the most frequent toxicity. Eighteen of 21 patients (86%) experienced grade 3 or 4 neutropenia during the initial treatment cycle. The median neutrophil nadir was 0.5 x 10(3)/microl (range, 0.1-1.6 x 10(3)/microl), and the median platelet nadir was 127 x 10(3)/microl (range, 18-460 x 10(3)/microl). Other toxicities more than grade 2 included malaise (two patients), and anorexia, infection, fever, pulmonary, and cardiac in one patient each. There were no objective responses, and 18 patients had stable disease for a median of 74 days. The median survival for all patients was 230 days, with 23% alive at 1 year. Topotecan as administered in this trial is reasonably well tolerated; however, the response rate was insufficient to warrant additional study in pleural mesothelioma. PMID- 9856667 TI - Low sensitivity of computed tomography in the staging of gastric lymphomas of mucosa-associated lymphoid tissue: impact on prospective trials and ordinary clinical practice. AB - The natural history and management of gastric lymphomas of mucosa-associated lymphoid tissue (MALTomas) are not completely understood. Most stage I cases are now entered into prospective trials to confirm the excellent results obtained with conservative treatment, whereas current therapeutic policies are based on accumulated experience. The limits of staging work-ups may have a significant impact on prospective trials and ordinary clinical practice. The authors explore the sensitivity of computed tomography scanning in detecting perigastric adenopathy in 20 patients with gastric MALToma treated by gastrectomy. Clinical staging identified 17 patients as having stage I MALTomas and three patients as having stage II1 MALTomas. Histopathologic staging showed that 8 of 17 patients formerly diagnosed with stage I MALToma had perigastric nodal involvement, whereas the three patients with clinical stage II1 were confirmed as such. Computed tomography scanning has low sensitivity in detecting perigastric lymphadenopathy in gastric MALTomas. This leads to understaging, with a significant impact on therapeutic decision, and distorts newly acquired knowledge about the disease's natural history and management, introducing a bias in prospective clinical trials. Endoscopic ultrasonography should be tested as a staging procedure both in prospective trials and in ordinary clinical practice. PMID- 9856668 TI - Brief courses of palliative radiotherapy for metastatic bone pain: a pilot cost minimization comparison with narcotic analgesics. AB - The use of radiotherapy to treat metastatic bone pain is being challenged by claims of high cost and by more readily available, noninvasive treatment approaches. The authors assessed the effectiveness of brief courses of radiotherapy in reducing pain and estimated cost data for a pilot comparison between radiotherapy and narcotic analgesics in patients with cancer. A representative group of outpatients undergoing brief courses of radiotherapy with Karnofsky scores above 70 and without serious comorbidities were recruited from 1995 through 1996. Patients indicated their pain at rest and with movement on a scale of from 1 to 10 both before and up to 12 months after radiotherapy. Radiotherapy costs were estimated from Medicare-allowable charges. Narcotic analgesia costs were estimated from published values. In 66 patients with 131 individually treated sites, median at rest pain score decreased by about 4 points after treatment (5.58 [-/+3.28] before treatment vs. 1.55 [-/+1.8] after treatment; p < 0.05). Median with movement pain score was about 5 points lower after treatment (7.32 [-/+2.72] before treatment vs. 1.94 [-/+2.07] after treatment; p < 0.05). No differences were found when stratifying by type of pain, tumor histologic type, or skeletal site. The estimated cost per patient ranged from $1,200 to $2,500 for radiotherapy. This compares with an estimated cost of $9,000 to $36,000 for 9 months of narcotics. In this pilot study, a brief course of radiotherapy significantly reduced pain and appeared to be cost effective when compared with narcotic analgesia. A full economic evaluation is warranted. PMID- 9856669 TI - Laryngeal cancer in patients with acquired immunodeficiency syndrome. PMID- 9856670 TI - The complete control and possible cure of a recurrent carcinoma of the breast by means of hormonal therapy over a 22-year period: a case report. PMID- 9856671 TI - Tumor cell apoptosis present at diagnosis may predict treatment outcome for patients with medulloblastoma. AB - PURPOSE: To determine if the degree of tumor cell apoptosis at diagnosis predicts outcome, tissue sections of medulloblastoma were examined and the amount of apoptosis and progression-free survival were correlated. PATIENTS AND METHODS: The study cohort consisted of 43 children in whom medulloblastoma was diagnosed between 1984 and 1995: 29 patients at high risk (HR) treated with radiation and chemotherapy, and 14 children at low risk (LR) treated with radiation alone. A terminal deoxynucleotidyl transferase (TdT) end-labeling assay was used to detect apoptosis in paraffin-embedded tissue sections prepared at diagnosis. RESULTS: Progression-free survival was examined in cohorts of children whose tumors were divided into quartiles based on the apoptotic index (AI) of their pretreatment tumor specimens. A comparison of these four groups of children revealed an association between AI and outcome (p = 0.03); patients with tumors in the highest AI quartile had substantially improved outcome compared to all other patients combined (p = 0.02). In this cohort of patients treated with different therapies, assignment at the time of diagnosis to LR and HR groups based on widely-accepted clinical criteria was not closely associated with outcome (p = 0.47). CONCLUSION: AI is a strong indicator of treatment outcome for children with medulloblastoma after treatment with cytotoxic therapy, independent of risk group. Because HR and LR patients included in this study received different modalities of cytotoxic therapy, it is possible that AI predicts outcome independent of the precise antineoplastic therapy a patient receives. PMID- 9856672 TI - The costs of children with sickle cell anemia: preparing for managed care. AB - PURPOSE: To anticipate the clinical challenges and financial risks facing physicians and managed care organizations who care for children with chronic illnesses, such as sickle cell anemia (SCA), under capitated managed care arrangements. PATIENTS AND METHODS: A cross-sectional study based on claims data from the Washington State Medicaid Program (WSMP) and the Federal Employees Health Benefits Program (FEP). Expenditure patterns were compared for children 18 years of age or younger for whom a claim with a diagnosis of SCA was submitted and paid in the State of Washington during fiscal year 1993 (FY1993) or by the FEP during FY1992 to expenditure patterns for all children. RESULTS: Children with SCA had mean expenditures 8.8 times the mean expenditures for all children in WSMP. There was wide variation in the annual expenditures among children with SCA; the most expensive 10% of children accounted for 56% of total expenditures. Ninety-seven percent of the expenditures were concentrated in four broad categories: 72% for inpatient care, 11% for outpatient care, 11% for physician payments, and 3% for prescription drugs. Examination of expenditure and utilization patterns for children with sickle cell anemia enrolled in the FEP yielded similar results. CONCLUSIONS: Unless managed care organizations and capitated pediatricians receive payment rates that reflect the higher expected expenditures of caring for these children, access to and quality of care may suffer. Analyses of practice guidelines and utilization patterns suggest that newborn screening, regular access to specialty facilities, and comprehensive education programs are critical areas that are vulnerable to reductions under capitation. PMID- 9856673 TI - No evidence for an effect of nutritional status at diagnosis on prognosis in children with acute lymphoblastic leukemia. AB - PURPOSE: To test the hypothesis that nutritional status at diagnosis, defined as body mass index standard deviation score (SDS), is related to the prognosis in childhood acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: The sample consisted of 1,025 patients with standard risk ALL who had been randomized to different intensification therapies. Outcome measures were relapse/no relapse and time to first relapse. The influence of body mass index SDS was tested by survival analysis. RESULTS: There was no evidence that body mass index SDS was related to clinical outcome (proportional hazards model, p = 0.72). CONCLUSIONS: The study results suggest that nutritional status at diagnosis, defined on the basis of the body mass index, at least in developed countries, has no effect on the prognosis in ALL, and it should not be considered as a prognostic factor. PMID- 9856674 TI - Erythroid marrow activity and hemoglobin H levels in hemoglobin H disease. AB - PURPOSE: To determine serum immunoreactive erythropoietin (Epo) and soluble transferrin receptors (sTfR) levels in patients with hemoglobin H (HbH) disease and the correlation with HbH levels and alpha-globin genotype. PATIENTS AND METHODS: Twenty patients with HbH disease were studied. Methods applied included cation-exchange high pressure liquid chromatography for HbH levels, chemoluminescence for Epo concentration, immunoassay for sTfR concentration, and DNA analysis for alpha-globin genotype characterization. RESULTS: Serum Epo and sTfR levels were significantly elevated (46.6+/-26.8 IU/l and 5.6+/-1.8 mg/l, respectively) in patients with HbH disease compared to controls (9.2+/-3.3 IU/l and 1.8+/-0.7 mg/l, respectively). Epo and sTfR levels correlated positively with HbH concentration (r = 0.93 and 0.80, respectively). The highest Epo and sTfR values were observed in three patients with the highest HbH levels who all had nondeletion alpha-thalassemia mutations. CONCLUSION: Epo and sTfR levels are increased in patients with HbH disease; this increase is directly related to the HbH concentration that usually reflects the degree of globin polypeptide imbalance. The correlation of Epo, sTfR, and reticulocyte production index in these patients indicates that anemia in HbH disease mainly is caused by ineffective erythropoiesis and a mild degree of peripheral hemolysis. PMID- 9856675 TI - Arsenic trioxide in the treatment of a patient with multiply recurrent, ATRA resistant promyelocytic leukemia: a case report. AB - PURPOSE: Little experience exists with the use of arsenic trioxide in the treatment of recurrent, all-trans retinoic acid (ATRA)-resistant, acute promyelocytic leukemia (APL). The authors report a patient with multiply recurrent APL treated with arsenic trioxide (As2O3), which was administered as recommended in the protocol from the People' s Republic of China. The results of this treatment and its toxicity are discussed. The available literature on arsenic therapy is reviewed. PATIENTS AND METHODS: The patient was a 15-year-old African-American girl with APL that had resisted conventional chemotherapy, ATRA therapy followed by autologous peripheral stem cell transplant, and a second course of ATRA induction therapy administered for relapse after transplant. The patient was treated with 10 mg As2O3 intravenously for 28 days. After a 4-week break, she received a second 28-day course of As2O3 therapy. RESULTS: After completion of the first 28-day course of As2O3 treatment, morphologic and cytogenetic remission occurred. Reverse-transcription polymerase chain reaction demonstrated persistence of the PML-RARalpha fusion transcript. After the second course of As2O3, the patient had a complete remission by morphologic, cytogenetic, and molecular criteria. Approximately 6 months after the end of two courses of As2O3 therapy, the patient again underwent relapse. An additional course of As2O3 achieved a morphologic, although not a cytogenetic or molecular, remission. CONCLUSIONS: As2O3 therapy produced remission in a patient with multiply relapsed, ATRA-resistant APL. Toxic side effects were minimal. The patient underwent relapse 6 months after this therapy. Further investigation will be necessary to determine the proper role of As2O3 therapy in patients with APL. PMID- 9856676 TI - T-cell acute lymphoblastic leukemia after renal transplantation in childhood. AB - PURPOSE: Lymphoproliferative disorders in solid organ recipients are usually of B cell type and have rarely been described in childhood. This study describes the development of T-cell acute lymphoblastic leukemia (ALL) in a child occurring 6 years after renal transplantation. PATIENT: An 11-year-old boy had received a renal allograft from his father at 5 years of age. He was receiving imuran, prednisone, and cyclosporin A prophylaxis for graft rejection after transplant until T-cell ALL was diagnosed. Although an acute Epstein-Barr virus (EBV) infection was noted at the time of diagnosis, the EBV genome was not detected by Southern blot analysis and polymerase chain reaction (PCR) in the leukemic cells. RESULTS: A large mediastinal mass and malignant pleural effusion were noted at diagnosis. Leukemic cells of his bone marrow and pleural fluid expressed T-cell antigens with unique cytogenetic features, including add(1)(p36.1), del(11)(q14), and monosomy 7. EBV serology was consistent with a recent infection but EBV genome was not detected by Southern blot and PCR analysis in his leukemic cells. Human T-lymphotropic virus-I (HTLV-I) antibody titer was negative. He has been on chemotherapy for 9 months, maintaining his first remission. CONCLUSIONS: Malignancies developing after renal transplantations are usually lymphoproliferative disorders and of B-cell origin. In the majority of these patients, EBV plays an etiologic role. Although adult T-cell leukemia developing during immunosuppressive treatment in renal transplant recipients has been reported, T-cell leukemia after transplant in pediatric patients has not been reported to date. This case is unique in terms of the patient's age, the T-cell immunophenotype, the cytogenetic features, and the absence of an EBV genome within the leukemic cells despite an acute EBV infection before diagnosis. PMID- 9856677 TI - Pediatric primary diffuse large cell lymphoma of bone with t(3;22)(q27;q11). AB - PURPOSE: A child with a primary lymphoma of bone (PLB) with a t(3;22)(q27;q11) is described. METHODS: An 11-year-old boy had a 5-week history of back pain and a destructive lesion of S1 that contained an epidural component. Histologic evaluation of a biopsy confirmed the diagnosis of diffuse large B-cell non Hodgkin lymphoma. Karyotypic analysis disclosed a t(3;22)(q27;q11), but the amount of tumor tissue was insufficient for molecular studies of the BCL-6 gene. RESULTS: The patient remains free of disease 4 years after completion of intensive systemic chemotherapy and intrathecal chemotherapy. CONCLUSIONS: The lymphoma in the patient described in this report is highly unusual because of the coexistence of pediatric PLB and a t(3;22)(q27q11). PMID- 9856678 TI - CD30-positive anaplastic large cell lymphoma (ALCL) of T-cell lineage in a 14 month-old infant with perinatally acquired HIV-1 infection. AB - PURPOSE: CD30-positive anaplastic large cell lymphoma (ALCL) has been described in adults with HIV-1 infection but is extremely rare in HIV-1-infected infants and children. PATIENT AND METHODS: A 14-month-old girl with congenitally acquired HIV-1 infection presented with fever and a tender, erythematous, cystic mass on the right labium majorum. The mass was biopsied. Histologic examination and immunohistochemistry were performed. RESULTS: Histologic examination showed Touton-like giant cells resembling histiocytes and focally abundant neutrophils obscuring the lymphoid infiltrate. Immunocytochemistry revealed a CD30-positive ALCL of T-cell lineage. CONCLUSION: Although non-Hodgkin's lymphoma is known to be associated with HIV-1 infection in children, large cell lymphomas of T-cell lineage are extremely rare in this population. Early diagnosis should be aggressively pursued in an HIV-1-infected child who presents with a fever and cutaneous mass. PMID- 9856679 TI - Congenital erythrocytosis with elevated erythropoietin level: an incorrectly set "erythrostat"? AB - PURPOSE: A child who was extensively evaluated for polycythemia is reported. Polycythemia, or erythrocytosis, is seen rarely in children. The mechanisms for congenital and/or familial erythrocytosis are discussed. PATIENT AND METHODS: A 10 1/2-year-old white girl was referred for evaluation of polycythemia, which was detected incidentally during an emergency room visit for a febrile illness. She underwent extensive evaluation to determine the cause of the polycythemia. The literature was reviewed to determine the occurrence of congenital and/or familial erythrocytosis in children and its various causes. RESULTS: Despite extensive evaluation, no specific cause of the erythrocytosis could be determined in our patient. The erythrocytosis appeared to be secondary to an inappropriately elevated serum erythropoietin concentration. Serum erythropoietin rose further after phlebotomy, suggesting nonautonomous hypersecretion. After a review of the literature, we hypothesize that she had an inappropriate erythropoietin expression related to an abnormality in the renal oxygen-sensing mechanism governing erythropoietin synthesis. DISCUSSION: A discussion of congenital and familial erythrocytosis is presented, and a review of the literature regarding the possible mechanisms causing erythrocytosis is included. PMID- 9856680 TI - Hb Hammersmith [beta 42(CD1) Phe-->Ser]: occurrence as a de novo mutation in black monozygotic twins with multiple congenital anomalies. AB - PURPOSE: To present the occurrence of Hb Hammersmith as a de novo mutation in African-American twins with multiple congenital anomalies. METHODS: Standard hematologic methods were used. The presence of an unstable Hb variant was confirmed by brilliant cresyl blue staining and an isopropanol stability test. Hb Hammersmith was confirmed by the sequencing of polymerase chain reaction amplified beta-globin gene. RESULTS: The presence of Hb Hammersmith was confirmed in female monozygotic twins of African-American origin with congenital Heinz body hemolytic anemia and multiple congenital anomalies. The variant occurred as a de novo mutation in the twins. CONCLUSION: This report describes the occurrence of Hb Hammersmith [B42(CD1)Phe-->Ser] in African-American twins. As with the other reported cases, both twins were female. In addition to Heinz body hemolytic anemia, a low arterial O2 saturation in the proposita was shown by pulse oximetry. Multiple congenital anomalies involving various systems were also found in both twins. PMID- 9856682 TI - Persistent tachypnea in children: keep pulmonary embolism in mind. AB - PURPOSE: Tachypnea in children is associated with respiratory disorders and nonrespiratory disorders such as cardiac disease, metabolic acidosis, fever, pain, and anxiety. Pulmonary embolism is seldom considered by pediatricians as a cause of tachypnea. PATIENTS AND METHODS: Three children of various ages with persistent tachypnea are described: a girl after orthopedic surgery for kyphoscoliosis, a boy with nephrotic syndrome, and a neonate with Hirschsprung disease. Other causes of tachypnea were diagnosed and treated before pulmonary embolism was considered. RESULTS: Ventilation-perfusion scanning appeared to be highly probable for pulmonary embolism in these patients. Anticoagulant therapy was started. CONCLUSION: Pulmonary embolism should be kept in mind in children with tachypnea, especially when other risk factors for venous thromboembolism are present, to avoid delay in anticoagulant treatment and a fatal outcome. PMID- 9856681 TI - Kasabach-Merritt syndrome-associated kaposiform hemangioendothelioma successfully treated with cyclophosphamide, vincristine, and actinomycin D. AB - PURPOSE: The authors introduced a successful trial of a chemotherapy regimen to treat refractory thrombocytopenia and pain caused by a rare kaposiform hemangioendothelioma that had been refractory to corticosteroids. PATIENT AND METHOD: A 6-year-old Latina girl with diffuse petechiae, constant pain and edema in her right lower extremity, and severe thrombocytopenia was brought for treatment. Physical examination revealed a large mass in the right lower extremity. The surgical specimen resected earlier in Mexico showed kaposiform hemangioendothelioma, an extremely rare vascular tumor that is frequently associated with consumptive coagulopathy and lymphangiomatosis. Corticosteroid therapy in Mexico had been ineffective in controlling the tumor size, pain, or thrombocytopenia. The patient was brought to the United States because of a rapidly enlarging tumor and intractable leg pain, causing inability to ambulate. Laboratory examinations revealed profound thrombocytopenia and evidence of consumptive coagulopathy. Because of the rapidly enlarging lesion, it was decided to give six cycles of a chemotherapy regimen consisting of vincristine, cyclophosphamide, and actinomycin D (VAC). RESULT: The patient responded to the first cycle of chemotherapy with partial reversal of the thrombocytopenia. The additional five cycles of the same chemotherapy regimen resulted in complete resolution of thrombocytopenia and significant improvement of the pain and edema in the right lower extremity. A follow-up imaging study 9 months later revealed a marked decrease in size of the vascular tumor. CONCLUSION: Inoperable kaposiform hemangioendothelioma unresponsive to corticosteroids can be successfully treated with chemotherapy. PMID- 9856683 TI - Solitary plasmocytoma of bone in an adolescent. AB - PURPOSE: To describe an adolescent with solitary plasmocytoma of bone and compare the pathologic and radiographic findings with osteomyelitis. PATIENT AND METHODS: A 17-year-old girl had a 3-year history of swelling of the right tibia, local pain, and hyperemia. RESULTS: Initial biopsy was interpreted as osteomyelitis. The lesion was rebiopsied 3 years later and showed a plasma cell neoplasm. A review of the first biopsy confirmed a similar histologic picture. CONCLUSION: Although solitary plasmocytoma of bone is a rare neoplasm in adolescence, it must be considered in the differential diagnosis of chronic osteomyelitis. PMID- 9856684 TI - Ewing's sarcoma in a patient with congenital optic atrophy. AB - PURPOSE: The authors report Ewing's sarcoma in a patient with congenital optic atrophy. Congenital optic atrophy is an autosomal dominant disorder with variable expression. It manifests itself within the first decade of life and is accompanied by changes in visual acuity. This condition has not been known to cause a predisposition toward malignancy. PATIENT AND METHODS: In this article, a 16-year-old girl with dominant optic atrophy and Ewing's sarcoma is described. RESULTS: After tumor resection, the patient was successfully treated for Ewing's sarcoma with chemotherapy and radiation therapy. Today she has no evidence of malignancy. However, she continues to have deterioration of her visual acuity. CONCLUSION: This report of cancer in a child with dominant optic atrophy suggests the possible need for a higher degree of suspicion of malignancy in these patients. PMID- 9856685 TI - Lipoblastoma presenting as a mesenteric mass in an infant. AB - PURPOSE: This article describes an infant with a large abdominal mass and hypertension. PATIENT AND METHODS: A 5-month-old infant girl with diarrhea of 1 week's duration and a large right-sided abdominal mass was brought for treatment. Computed tomography of the abdomen revealed a large, generally homogeneous, hypodense mass, which compressed the right kidney, resulting in dilatation of the right renal collecting system. At surgery, the mass was adherent anteriorly to the transverse colon and attached by a stalk to the mesentery near the origin of the right colic artery. RESULTS: Examination of the mass showed an encapsulated lipoblastoma. Cytogenetic analysis revealed a 46,XX karyotype with a reciprocal translocation between chromosome 2 and chromosome 8 with breakpoints at q23 and q11.2, respectively. CONCLUSION: Lipoblastoma is a rapidly growing but benign tumor, which can cause severe medical problems by compressing major organs. Cytogenetic analysis can reveal translocations involving chromosome 8 band q11.2, which appears to be a specific chromosome marker for lipoblastoma. PMID- 9856686 TI - Multiple pulmonary chemodectomas in a child: results of four different therapeutic regimens. AB - PURPOSE: Chemodectomas (or paragangliomas) are rare tumors of neuroendocrine chemoreceptors, such as the carotid body. This report describes a case of multiple pulmonary chemodectomas in an adolescent and discusses the results of four therapeutic regimens. PATIENTS: At 15 years of age, the patient had cough and fatigue. Investigation revealed numerous 1- to 2-cm diameter nodules throughout both lungs. Biopsy revealed multiple pulmonary chemodectomas of uncertain malignant potential. No extrapulmonary primary site could be found. RESULTS: Because of deteriorating pulmonary function, she was treated with courses of etoposide-cisplatin and subsequently somatostatin without effect. She finally responded to a course of doxorubicin and streptozocin. She is currently maintained on interferon-alpha 2B but her measured vital capacity continues to fall slowly, reflecting increased tumor growth. Because there is still no evidence of extrapulmonary spread, she is considered to be a candidate for lung transplantation. CONCLUSIONS: A doxorubicin-streptozocin combination produced a temporary remission of this patient's multiple pulmonary chemodectomas. PMID- 9856687 TI - Intrathecal vincristine: an analysis of reasons for recurrent fatal chemotherapeutic error with recommendations for prevention. AB - PURPOSE: Accidental intrathecal vincristine instillation is usually a fatal error. The authors report an analysis of a patient and suggest means with which to reduce such errors. PATIENTS AND METHODS: A 7-year-old girl with recurrent acute lymphoblastic leukemia was inadvertently injected intrathecally with 1.5 mg vincristine. A detailed analysis of the events leading to this error and a review of all reported cases in the English literature were undertaken. RESULTS: Reasons for errors reported by us and other institutions included mistaking vincristine for an intended intrathecal drug, assuming vincristine was an additional drug to be injected, not checking physician orders, mistaken route of administration, and mislabeling of syringes. CONCLUSION: Intrathecal injection of vincristine may be the end-result of a series of systems errors. Protocol recommendations to reduce the likelihood of this error are presented. PMID- 9856688 TI - Modulation of plasma metalloproteinase-9 concentrations and peripheral blood monocyte mRNA levels in patients with septic shock: effect of fiber-immobilized polymyxin B treatment. AB - The authors measured plasma metalloproteinase (MMP)-9 and corresponding monocyte mRNA in 20 patients with septic shock. Plasma MMP-9 concentrations and monocyte MMP-9 mRNA levels were significantly higher in the 10 nonsurviving patients with septic shock than in 10 surviving patients and 25 normal controls. Hemoperfusion using polymyxin B immobilized on fibers (PMX-F), a reportedly effective treatment for septic shock, was studied for effects on MMP-9 in the patients. Increases in plasma MMP-9 concentrations and corresponding monocyte mRNA levels were attenuated significantly by PMX-F treatment in both nonsurviving and surviving patients. These data suggest that plasma MMP-9 concentrations and monocyte MMP-9 mRNA levels may be useful prognostic markers in septic shock, and that PMX-F treatment affects MMP-9. PMID- 9856690 TI - Heparin inhibits lung branching morphogenesis: potential role of smooth muscle cells in cleft formation. AB - Lung branching morphogenesis is the process by which the embryonic lung undergoes repetitive branching to form the bronchial tree. This process occurs during the pseudoglandular stage of lung development and requires epithelial-mesenchymal interactions. Coinciding with lung branching morphogenesis is the appearance of parabronchial smooth muscle cells (PSMCs) and the accumulation of extracellular matrices (ECMs) around the developing airways. The authors previously reported in preliminary form that heparin prevents the branching of murine lung explants (Roman et al., Am Rev Respir Dis. 1991; 143:A401); this article corroborates those early observations and expands them by demonstrating that heparin results in disruption of PSMC distribution and abnormal organization of ECMs around the developing airways. These changes were associated with inhibition of lung branching morphogenesis in the absence of effects on cell proliferation. The data provide further support for the role of ECMs in lung branching morphogenesis, and points to PSMCs as potential players in this process. PMID- 9856689 TI - Differential impact of obesity on glucose metabolism in black and white American adolescents. AB - BACKGROUND: The authors have previously demonstrated abnormalities in glucose and insulin metabolism in nondiabetic black American (BA) adults versus white American (WA) adults. Whether similar glucoregulatory alterations extend to BA adolescents remain unknown. In addition, obesity, a known risk factor for insulin resistance and hyperinsulinemia, occurs in a greater proportion of BA adults and children when compared to WA. The objective of the present study was to examine the differential effects of obesity on glucose homeostasis in BA and WA adolescents. METHODS: We examined glucose homeostasis in BA and WA adolescents using oral glucose tolerance test (OGTT), intravenous glucose tolerance test (IVGTT), and [6,6-2H2]-glucose infusion. The study consisted of four age-, sex-, and pubertal stage-matched groups: 15 lean BA, 29 lean WA, 7 obese BA, and 9 obese WA. RESULTS: Both obese groups had significantly increased insulin and C peptide area under the curve (AUC) during OGTT and IVGTT when compared to their same-race lean counterparts. During OGTT, obese BA demonstrated greater insulin and C-peptide when compared to obese WA. During IVGTT, first- and second-phase insulin were significantly greater in obese BA versus obese WA. CONCLUSION: In summary, BA adolescents demonstrated insulin resistance which is markedly exaggerated in the face of obesity when compared to WA adolescents, implying a differential impact for obesity on glucose homeostasis that is unique to the obese BA adolescent group. In conclusion, there is a need for early aggressive weight management in obese BA adolescents. PMID- 9856691 TI - Multiresistant Shigella species isolated from pediatric patients with acute diarrheal disease. AB - A total of 57 strains of Shigella (36 S sonnei, 21 S flexneri), isolated from children with acute diarrheal disease who presented for treatment at the Andes University Hospital, Merida, Venezuela, from June 1993 to June 1995, were tested for their susceptibility to trimethoprim, sulfamethoxazole, ampicillin, cefamandole, ceftriaxone, streptomycin, fleroxacin, and nalidixic acid, by the agar dilution method. Twenty-seven strains (75%) of S sonnei and eight strains of S flexneri (38.1%) isolates showed high-level resistance to trimethoprim (MIC90 > 1024 microg/mL), which was also associated with other resistance patterns. The most common resistant phenotype associated with trimethoprim-resistance among S sonnei isolates was sulfamethoxazole-streptomycin (63%); among S flexneri isolates, it was sulfamethoxazole-ampicillin-streptomycin (87.5%). Individual resistance was only observed for ampicillin, mainly in four isolates of S flexneri, and in one isolate of S sonnei. Most Shigella strains were resistant to three or more antimicrobial agents. These results confirmed that multiresistant strains of Shigella are present in Merida, and emphasize the importance to maintain these under surveillance in order to assess local susceptibility patterns and empiric therapy. PMID- 9856692 TI - A patient with recurrent ascites. PMID- 9856693 TI - Amiodarone-associated hemoptysis. AB - Amiodarone is a benzofuran derivative with a chemical structure similar to thyroxine. Originally introduced to treat angina pectoris, amiodarone was found to have antiarrhythmic properties, and in 1985, was approved in the United States for treatment of life-threatening ventricular arrhythmias. It is now used for various ventricular and supraventricular arrhythmias refractory to conventional first-line medications, and as a result, side effects have been observed with increased frequency. The most severe and potentially life-threatening of these side effects is the development of pulmonary toxicity. Typically, amiodarone pulmonary toxicity (APT) is manifested by acute pneumonitis and chronic fibrosis. Amiodarone-associated hemoptysis (AAH) is a rare occurrence. The authors describe a case of AAH successfully treated with cessation of drug and steroid therapy. PMID- 9856694 TI - Hyperglobulinemia with an increased anion gap. AB - Plasma proteins, especially albumin, account for much of the normal anion gap. The relationship between globulins and the anion gap has not received much attention except for reports of a narrowing of the gap associated with certain monoclonal immunoglobulin G gammopathies. The author presents a patient with polyclonal gammopathy, the magnitude of which correlated strongly and positively with the anion gap, along with data from 400 patients attending a renal hypertension clinic, which show a significant positive correlation between the anion gap and total globulins. These results indicate that both albumin and globulin concentrations should be taken into account when attempting to explain abnormalities or changes in the magnitude of the gap. PMID- 9856695 TI - Long-lasting response to vinorelbine in unresectable non-small cell lung carcinoma: a case with concomitant good quality of life. AB - The authors observed a long-lasting response to uninterrupted vinorelbine treatment in a 72-year-old patient with a stage IIIB unresectable non-small cell carcinoma. The progressive tumor mass reduction was assessed by computed tomographic scans over a 2-year period. In the literature, data of randomized trials confirm that the appeal of single-agent vinorelbine therapy in elderly patients is also good for the patient's quality of life during treatment. PMID- 9856696 TI - Hematologic abnormalities associated with influenza A infection: a report of 3 cases. AB - Influenza A is associated with leukopenia, although it is not reported to cause isolated thrombocytopenia, anemia, or pancytopenia. The authors report three pediatric patients with transient cytopenias associated with influenza A infection, all of whom had evidence for influenza A infection by direct immunofluorescence from nasopharyngeal aspirates. In all patients, cytopenias were transient and improved as their viral symptoms resolved. All patients improved spontaneously. This is the first report of transient pancytopenia, anemia, or thrombocytopenia associated with influenza A infection. Given the high frequency of influenza A infections during the winter months, it is important to recognize the associated hematologic findings. PMID- 9856697 TI - Successful acyclovir treatment of herpes simplex type 2 hepatitis in a patient with systemic lupus erythematosus: a case report and meta analysis. AB - Herpes simplex virus (HSV) hepatitis is a rare complication of HSV infection with a high reported mortality rate in untreated patients. The authors present a case of HSV hepatitis in a 26-year-old female with focal proliferative lupus nephropathy who was status post one cycle of pulse high-dose (1 gm/ m2) cyclophosphamide. Treatment with parenteral acyclovir was successful. A meta analysis of well-documented cases of HSV hepatitis treated with acyclovir, excluding those that omit initial serum concentrations of hepatic transaminases, suggests that the early administration of parenteral acyclovir may have been instrumental in the achievement of a successful outcome, and that a patient's serum levels of hepatic transaminases at the time of treatment initiation may predict outcome. This is the first reported case of successful parenteral acyclovir treatment of HSV hepatitis in a patient with lupus nephritis who has recently undergone cyclophosphamide immunosuppression, and includes a meta analysis to examine the hypothesis that initial markers of hepatic injury may predict outcome of acyclovir treatment. PMID- 9856698 TI - Headaches, idiopathic intracranial hypertension, and pseudopapilledema. AB - A young woman with frontal headaches of several months' evolution and monocular transient "tunnel" obscurations that developed after a generalized seizure is described. She had elevation of the optic discs (pseudopapilledema), greater on the side of her visual symptoms. No intracranial lesions were identified. Cerebrospinal fluid pressure was increased, diagnostic of idiopathic intracranial hypertension (IIH). Because both IIH and pseudopapilledema may cause transient visual obscurations (TVO), fortuitous discovery of pseudopapilledema in a patient with TVO does not preclude the necessity of performing a diagnostic lumbar puncture. Establishing the correct diagnosis has obvious therapeutic and prognostic implications. PMID- 9856699 TI - Urinary sediment examination and Gulf War Syndrome. PMID- 9856700 TI - Urinary sediment examination and Gulf War Syndrome. PMID- 9856701 TI - Plagiarism. PMID- 9856702 TI - Assessing fluid responsiveness by the systolic pressure variation in mechanically ventilated patients. Systolic pressure variation as a guide to fluid therapy in patients with sepsis-induced hypotension. PMID- 9856703 TI - Blood volume measurement: the next intraoperative monitor? PMID- 9856704 TI - Systolic pressure variation as a guide to fluid therapy in patients with sepsis induced hypotension. AB - BACKGROUND: Monitoring left ventricular preload is critical to achieve adequate fluid resuscitation in patients with hypotension and sepsis. This prospective study tested the correlation of the pulmonary artery occlusion pressure, the left ventricular end-diastolic area index measured by transesophageal echocardiography, the arterial systolic pressure variation (the difference between maximal and minimal systolic blood pressure values during one mechanical breath), and its delta down (dDown) component (= apneic - minimum systolic blood pressure) with the response of cardiac output to volume expansion during sepsis. METHODS: Preload parameters were measured at baseline and during graded volume expansion (increments of 500 ml) in 15 patients with sepsis-induced hypotension who required mechanical ventilation. Each volume-loading step (VLS) was classified as a responder (increase in stroke volume index > or = 15%) or a nonresponder. Successive VLSs were performed until a nonresponder VLS was obtained. RESULTS: Thirty-five VLSs (21 responders) were performed. Fluid loading caused an overall significant increase in pulmonary artery occlusion pressure and end-diastolic area index, and a significant decrease in systolic pressure variation and delta down (P < 0.01). There was a significant difference between responder and nonresponder VLSs in end-diastolic area index, systolic pressure variation, and dDown, but not in pulmonary artery occlusion pressure. Receiver operator curve analysis showed that dDown was a more accurate indicator of the response of stroke volume index to volume loading than end-diastolic area index and pulmonary artery occlusion pressure. A dDown component of more than 5 mmHg indicated that the stroke volume index would increase in response to a subsequent fluid challenge (positive and negative predictive values: 95% and 93%, respectively). CONCLUSION: The dDown component of the systolic pressure variation is a sensitive indicator of the response of cardiac output to volume infusion in patient with sepsis-induced hypotension who require mechanical ventilation. PMID- 9856705 TI - Blood volume measurement at the bedside using ICG pulse spectrophotometry. AB - BACKGROUND: In the treatment of critically ill patients, blood volume (BV) measurement requires injection of some tracer substance and subsequent blood sampling to analyze the tracer concentration. To obviate both the sampling and laboratory analysis, techniques of pulse oximetry have been adapted to the noninvasive optical measurement in the patient's nose or finger of the arterial concentration of an injectable dye. METHODS: The authors report the clinical accuracy of a new noninvasive bedside BV measurement test that uses pulse spectrophotometry (the pulse method). The device detects pulsatile changes of tissue optical density of a nostril or a finger spanned by a probe emitting two infrared wavelengths (805 and 890 nm). After a peripheral or central intravenous injection of indocyanine green, the arterial dye concentration is continuously computed by reference to the previously measured blood hemoglobin concentration. Three types of tests of its accuracy are described here. RESULTS: In 10 healthy volunteers, the authors compared BV determined by the pulse method with an (131)I labeled human serum albumin method. Three subject data sets were excluded because of motion artifact, a low signal:noise ratio, or both. For the other seven volunteers, the bias+/-SD of pulse spectrophotometric BV values were 0.20+/-0.24 l (or 4.2+/-4.9%) for the nose probe and 0.34+/-0.31 l (or 7.3+/-6.9%) for the finger probe, with a mean BV of 5 l. In 30 patients who underwent cardiac surgery, the pulse method was compared with a standard indocyanine green method using intermittent blood sampling. In three patients, the BV could not be determined by the pulse method because of motion artifact, low signal:noise ratio, or both. In 27 patients, the bias+/-SD of the BV by the pulse method was 0.23+/-0.37 l (-5.3+/-8.7%) for the nose and -0.25+/-0.5 l (-4.2+/-8.4%) for the finger. Patient BV ranged from 2.51 to 7.13 l (mean, 4.48 l). In 10 additional patients before cardiac surgery, BV was measured by the pulse method before and shortly after removal of 400 ml blood. The pulse method recorded a decrease of BV of 480+/-114 mL Three days after venesection, the mean BV was 117+/-159 ml less than the predonation control. CONCLUSIONS: In most patients, the pulse method provides bedside measurement of BV without blood sampling (except for hemoglobin determination), with an estimated error less than 10%. In 10-30% of tests the method failed because of motion distortion of the record during the 10-min data collection period or because of insufficient pulse amplitude in the test tissue. PMID- 9856706 TI - Circulating blood volume measured by pulse dye-densitometry: comparison with (131)I-HSA analysis. AB - BACKGROUND: Pulse dye-densitometry (PDD) is a newly developed technique for monitoring the arterial concentration of indocyanine green. Using this method, circulating blood volume (CBV) can be calculated without using radioisotopes. In this study, the CBV value obtained by PDD was validated by comparison using the human serum albumin ((131)I-HSA) dilution method. METHODS: Eleven healthy volunteers underwent placement of cannulae into the radial artery and antecubital vein for withdrawal of blood samples and injection of indicator. Probes for PDD were attached to the right nostril and the right index finger. Indocyanine green (20 mg), dissolved in 4 ml water, and 25 microCi (131)I-HSA in 1 ml distilled water were injected simultaneously into the left antecubital vein. Blood samples were withdrawn 3, 6, 10, 20, 30, and 45 min after injection, then processed for spectrophotometric measurement of indocyanine green and scintillation counting. RESULTS: The blood dye concentration correlated well with the values obtained by PDD (r=0.986, imprecision 0.04+/-0.11 mg/l, 10.0+/-31%. The imprecision of the CBV value obtained by PDD (nose probe) and by the (131)I-HSA dilution method was 3.99+/-10.54%, 0.259+/-0.593 l. The imprecision of the CBV obtained by in vitro spectrophotometry compared with PDD was 2.47+/-9.00%, 0.100+/-0.446 l. CONCLUSIONS: This newly developed, less invasive method can measure CBV with an imprecision of 3.99+/-10.54%, 0.259+/-0.593 l (nose probe), and thus is also as accurate as the conventional radioisotope method. PMID- 9856707 TI - A randomized study of combined spinal-epidural analgesia versus intravenous meperidine during labor: impact on cesarean delivery rate. AB - BACKGROUND: Combined spinal-epidural (CSE) analgesia produces rapid-onset pain relief and allows ambulation in early labor. Epidural local anesthetics may contribute to an increase in operative deliveries by decreasing perineal sensation and causing motor weakness. Operative delivery rates might be reduced with CSE, by avoiding or delaying administration of local anesthetics. This study compares the operative delivery rates associated with a CSE technique and those associated with intravenous meperidine for labor analgesia. METHODS: Healthy parturients at full term were assigned randomly to receive CSE or intravenous meperidine analgesia. The CSE group received 10 microg intrathecal sufentanil, followed by epidural bupivacaine and fentanyl at their next request for analgesia. Parturients receiving intravenous meperidine had 50 mg on demand (maximum, 200 mg in 4 h). Labor and delivery outcomes in both groups were recorded and compared. RESULTS: An intent-to-treat analysis of 1,223 women indicated that CSE does not increase the rate of cesarean delivery for dystocia in nulliparous and parous women (CSE, 3.5% vs. intravenous meperidine, 4; P=not significant) or in nulliparous women alone (CSE, 7% vs. intravenous meperidine, 8%; P=not significant). Profound fetal bradycardia that necessitated emergency cesarean delivery within 1 h of the time the mother received sufentanil occurred in 8 of 400 parturients (compared with 0 of 352 who received meperidine; P < 0.01). However, the method of fetal monitoring differed between the two groups. Despite this, neonatal outcomes were similar overall. CONCLUSIONS: Combined spinal-epidural analgesia during labor does not increase the cesarean delivery rate for dystocia in healthy parturient patients at full term, regardless of parity. However, an unexpected increase in the number of cesarean deliveries for profound fetal bradycardia after intrathecal sufentanil was observed. Further investigation is warranted. PMID- 9856708 TI - Low-dose lidocaine suppresses experimentally induced hyperalgesia in humans. AB - BACKGROUND: The antinociceptive effects of systemically administered local anesthetics have been shown in various conditions, such as neuralgia, polyneuropathy, fibromyalgia, and postoperative pain. The objective of the study was to identify the peripheral mechanisms of action of low-dose local anesthetics in a model of experimental pain. METHODS: In a first experimental trial, participants (n=12) received lidocaine systemically (a bolus injection of 2 mg/kg in 10 min followed by an intravenous infusion of 2 mg x kg(-1) x h(-1) for another 50 min). In a second trial, modified intravenous regional anesthesia was administered to exclude possible central analgesic effects. In one arm, patients received an infusion of 40 ml lidocaine, 0.05%; in their other arm, 40 ml NaCl, 0.9%, served as a control. In both trials, calibrated tonic and phasic mechanical and chemical (histamine) stimuli were applied to determine differentially the impairment of tactile and nociceptive perception. RESULTS: Mechanical sensitivity to touch, phasic mechanical stimuli of noxious intensity, and heat pain thresholds remained unchanged after systemic and regional application of the anesthetic. In contrast, histamine-induced itch (intravenous regional anesthesia), axon reflex flare (systemic treatment), and development of acute mechanical hyperalgesia during tonic pressure (12 N; 2 min) of an interdigital web was significantly suppressed after both treatments. CONCLUSIONS: Increasing painfulness during sustained pinching has been attributed to excitation and simultaneous sensitization of particular Adelta- and C-nociceptors. This hyperalgesic mechanism seems to be particularly sensitive to low concentrations of lidocaine. These findings confirm clinical experience with lidocaine in pain states dominated by hyperalgesia. PMID- 9856709 TI - Bupivacaine 0.01% and/or epinephrine 0.5 microg/ml improve epidural fentanyl analgesia after cesarean section. AB - BACKGROUND: The authors studied the addition of bupivacaine and epinephrine, separately and together, to epidural fentanyl to determine whether this improved postcesarean analgesia and reduced the incidence of side effects. METHODS: After elective cesarean section, 100 parturient patients who received fentanyl (3 microg/ml) epidurally for 48 h were allocated randomly in a double-blinded manner to four groups to receive, in addition to the study solution, 0.01% bupivacaine, 0.5 microg/ml epinephrine, both, or neither. A neurologic assessment of breast fed neonates was made at 2 and 48 h of life. Plasma fentanyl concentrations were determined in a subset of patients at intervals after treatment. RESULTS: Patients receiving fentanyl alone made more attempts at patient-controlled analgesia (P < 0.01), required a greater total dose of fentanyl (P < 0.01), reported more pain (P < 0.003) and less satisfaction (P < 0.003), and had more nausea and urinary retention (P < 0.05) than all other groups. Patients who received bupivacaine with or without epinephrine had better overall satisfaction scores than those who did not receive bupivacaine (P < 0.001), and they required less fentanyl (P < 0.02) than patients who received fentanyl with only epinephrine. Motor blockade or orthostatic hypotension did not develop in any patient, and all patients could ambulate without difficulty. Neurobehavioral scores, which were similar among all neonates, were within the normal range. Plasma concentrations of fentanyl increased after epinephrine-containing solutions were discontinued. CONCLUSIONS: During the conditions of this study, the addition of epinephrine and bupivacaine to a 3-microg/ml epidural fentanyl solution for postcesarean section pain relief provided superior analgesia compared with fentanyl alone or fentanyl with epinephrine. Whether increasing the concentration of fentanyl alone might improve the efficacy of fentanyl remains unclear. PMID- 9856710 TI - Multicenter study of contaminated percutaneous injuries in anesthesia personnel. AB - BACKGROUND: Anesthesia personnel are at risk for occupational infection with bloodborne pathogens from contaminated percutaneous injuries (CPIs). Additional information is needed to formulate methods to reduce risk. METHODS: The authors analyzed CPIs collected during a 2-yr period at 11 hospitals, assessed CPI underreporting, and estimated risks of infection with human immunodeficiency virus and hepatitis C virus. RESULTS: Data regarding 138 CPIs were collected: 74% were associated with blood-contaminated hollow-bore needles, 74% were potentially preventable, 30% were considered high-risk injuries from devices used for intravascular catheter insertion or obtaining blood, and 45% were reported to hospital health services. Corrected for injury underreporting, the CPI rate was 0.27 CPIs per yr per person; per full-time equivalent worker, there were 0.42 CPIs/yr. The estimated average 30-yr risks of human immunodeficiency virus or hepatitis C virus infection per full-time equivalent are 0.049% and 0.45%, respectively. Projecting these findings to all anesthesia personnel in the United States, the authors estimate that there will be 17 human immunodeficiency virus infections and 155 hepatitis C virus infections in 30 yr. CONCLUSIONS: Performance of anesthesia tasks is associated with CPIs from blood-contaminated hollow-bore needles. Thirty percent of all CPIs would have been high-risk for bloodborne pathogen transmission if the source patients were infected. Most CPIs were potentially preventable, and fewer than half were reported to hospital health services. The results identify devices and mechanisms responsible for CPIs, provide estimates of risk levels, and permit formulation of strategies to reduce risks. PMID- 9856711 TI - Effect of various lithotomy positions on lower-extremity blood pressure. AB - BACKGROUND: Compartment syndrome of a lower extremity from hypoperfusion is a rare but potentially devastating complication of the lithotomy position during surgery. The aim of this study is to determine the effects of various lithotomy positions on lower-extremity blood pressures. METHODS: Blood pressure in eight young, healthy people was studied for 10 lithotomy positions. Blood pressure measurements were taken in both the upper arm (brachial artery) and the lower extremity (dorsalis pedis). The heart-to-ankle height gradient in each position was measured, and a predicted lower-extremity systolic pressure was calculated. The measured and predicted lower-extremity systolic blood pressures were compared with repeated measures analysis of variance. RESULTS: As a group, the mean systolic blood pressures in the lower extremities correlated closely with the predicted values. However, the difference between measured and predicted pressures varied among the 10 positions (P < 0.05). CONCLUSIONS: Although lower extremity systolic blood pressures in the young, healthy volunteers correlated with predicted values, there was an additional reduction in pressure associated with the lithotomy position. This surprising finding suggests that a lengthy procedure necessitating the use of a lithotomy position for only a portion should be planned so the remainder of the procedure can take place before establishing the position or so the position can be changed to an alternative position when it is no longer needed. PMID- 9856712 TI - Efficacy and costs of patient-controlled analgesia versus regularly administered intramuscular opioid therapy. AB - BACKGROUND: Many studies have shown the efficacy of patient-controlled analgesia (PCA). However, it is not clear whether PCA has clinical or economic benefits in addition to efficient analgesia. The current study was designed to evaluate these issues by comparing PCA with regularly administered intramuscular injections of opioids after hysterectomy. METHODS: This prospective study included 126 patients who underwent abdominal hysterectomy and were randomly assigned to receive PCA or regularly timed intramuscular injections of morphine during a period of 48 h. Doses were adjusted to provide satisfactory analgesia in both treatment groups. Pain at rest and with movement, functional recovery, drug side effects, and patient satisfaction were measured using rating scales and questionnaires. The costs of PCA and intramuscular therapy were calculated based on personnel time and drug and material requirements. RESULTS: Comparable analgesia was observed with the two treatment methods, with no significant differences in the incidence of side effects or patient satisfaction. The medication dosage had to be adjusted significantly more frequently in the intramuscular group than in the PCA patients. The PCA did not favor a faster recuperation time compared with intramuscular therapy in terms of times to ambulation, resumption of liquid and solid diet, passage of bowel gas, or hospital discharge. The results of the economic evaluation, which used a cost-minimization model and sensitivity analyses, showed that PCA was more costly than regular intramuscular injections despite the fact that no costs for the pump were included in the analyses. Cost differences in nursing time favoring PCA were offset by drug and material costs associated with this type of treatment. CONCLUSIONS: Compared with regularly scheduled intramuscular dosing, PCA is more costly and does not have clinical advantages for pain management after hysterectomy. Because of the comparable outcomes, the general use of PCA in similar patients should be questioned. PMID- 9856713 TI - Effects of hyperventilation and hypocapnic/normocapnic hypoxemia on renal function and lithium clearance in humans. AB - BACKGROUND: Using the renal clearance of lithium as an index of proximal tubular outflow, this study tested the hypothesis that acute hypocapnic hypoxemia decreases proximal tubular reabsorption to the same extent as hypocapnic normoxemia (hyperventilation) and that this response is blunted during normocapnic hypoxemia. METHODS: Eight persons were studied on five occasions: (1) during inhalation of 10% oxygen (hypocapnic hypoxemia), (2) during hyperventilation of room air leading to carbon dioxide values similar to those with hypocapnic hypoxemia, (3) during inhalation of 10% oxygen with the addition of carbon dioxide to produce normocapnia, (4) during normal breathing of room air through the same tight-fitting face mask as used on the other study days, and (5) during breathing of room air without the face mask. RESULTS: Hypocapnic and normocapnic hypoxemia and hyperventilation increased cardiac output, respiratory minute volume, and effective renal plasma flow. Glomerular filtration rate remained unchanged on all study days. Calculated proximal tubular reabsorption decreased during hypocapnic hypoxemia and hyperventilation but remained unchanged with normocapnic hypoxemia. Sodium clearance increased slightly during hypocapnic and normocapnic hypoxemia, hyperventilation, and normocapnic normoxemia with but not without the face mask. CONCLUSIONS: The results indicate that (1) respiratory alkalosis with or without hypoxemia decreases proximal tubular reabsorption and that this effect, but not renal vasodilation or natriuresis, can be abolished by adding carbon dioxide to the hypoxic gas; (2) the increases in the effective renal plasma flow were caused by increased ventilation rather than by changes in arterial oxygen and carbon dioxide levels; and (3) the natriuresis may be secondary to increased renal perfusion, but application of a face mask also may increase sodium excretion. PMID- 9856714 TI - Additive effect of nitric oxide inhalation on the oxygenation benefit of the prone position in the adult respiratory distress syndrome. AB - BACKGROUND: The response to inhaled nitric oxide and prone positioning was investigated in 47 patients with adult respiratory distress syndrome to test the hypothesis that inhalation of nitric oxide when in the prone position would result in additive improvement in oxygenation. METHODS: The authors prospectively studied patients of both genders who were 15 to 75 yr old and had adult respiratory distress syndrome confirmed by computed tomography (lung injury score, 3.1+/-1). RESULTS: Compared with baseline values in the supine position (T1), inhalation of 10 ppm nitric oxide for 1 h (T2) decreased the mean pulmonary artery pressure from 33+/-9 mmHg to 28+/-6 mmHg (P < 0.05; T2 vs. T1) and increased the ratio of the partial pressure of oxygen in arterial blood (PaO2) to inspired oxygen concentration (FiO2) from 115 (median first quartile [Q1] 97, median third quartile [Q3] 137) to 148 (Q1 132, Q3 196) (P < 0.05; T2 vs. T1). Cessation of nitric oxide brought the values back to baseline (T3). Two hours of prone positioning (T4) significantly increased the PaO2:FiO2 ratio (T4 vs. T3). However, after an additional hour of nitric oxide inhalation in the prone position (T5), a significant decrease of the venous admixture (from 33+/-6% to 25+/-6%; P < 0.05) and an increase of the PaO2:FiO2 ratio (from 165 [Q1 129, Q3 216] to 199 [Q1 178, Q3 316] [P < 0.05; T5 vs. T4]) were observed. CONCLUSIONS: In patients with isolated severe adult respiratory distress syndrome, inhalation of nitric oxide in the prone position significantly improved oxygenation compared with nitric oxide inhalation in the supine position or in the prone position without nitric oxide. The combination of the prone position with nitric oxide inhalation in the treatment of severe adult respiratory distress syndrome should be considered. PMID- 9856715 TI - Isoflurane and sevoflurane augment norepinephrine responses to surgical noxious stimulation in humans. AB - BACKGROUND: Suppression of hypertensive response to noxious stimulation by volatile anesthetics may be a result of suppression of the stimulation-induced norepinephrine response or that of the cardiovascular response to catecholamines, or both. The suppression of the cardiovascular response is established, but that of norepinephrine response has not been confirmed. The authors hypothesized that the suppression of cardiovascular response but not that of norepinephrine response plays a major role in suppressing the noxious stimulation-induced hypertensive response by volatile anesthetics. METHODS: Forty healthy donors for living-related liver transplantation were allocated to four groups: receiving 1.2% (end-tidal) isoflurane in oxygen and nitrogen, 2.0% isoflurane, 1.7% sevoflurane, or 2.8% sevoflurane. The intraoperative plasma norepinephrine and epinephrine concentrations, arterial blood pressure and pulse rate were measured for the first 15 min of surgery and were compared with the preoperative values. RESULTS: Norepinephrine and epinephrine concentrations both increased intraoperatively in all four groups. The values of maximum increase and area under the concentration-versus-time curve of norepinephrine were greater in the high dose groups of both anesthetics. The intraoperative blood pressure did not differ by different doses of anesthetics, and the degree of increase of blood pressure was not proportional to the plasma catecholamine concentrations. CONCLUSION: The effects of isoflurane and sevoflurane on the surgical noxious stimulation-induced norepinephrine response were inversely proportional to the dose. The suppression of noxious stimulation-induced blood pressure response by anesthetics that were studied may be the result of suppression of the responses of vascular smooth muscle and myocardium to catecholamines. PMID- 9856716 TI - The local addition of tenoxicam reduces the incidence of low back pain after lumbar epidural anesthesia. AB - BACKGROUND: Postepidural backache is a common postoperative complaint after lumbar epidural anesthesia. Useful interventions to decrease the incidence of postepidural backache would be helpful. METHODS: We performed a prospective, randomized, double-blind study to compare the effect of local addition of tenoxicam on the incidence of postepidural backache after nonobstetric surgery. One thousand unpremedicated ASA physical status I or II patients scheduled for hemorrhoidectomy were assigned randomly to tenoxicam or control groups. Patients in the control group received 25 ml lidocaine, 2%, with epinephrine 1:200,000 epidurally and 4 ml lidocaine, 1%, for local skin infiltration. Patients in the tenoxicam group received 25 ml lidocaine, 2%, with epinephrine 1:200,000 epidurally and 4 ml lidocaine, 1%, with tenoxicam (2 mg) 1:2,000 for local skin infiltration. Patients were interviewed at 24, 48, and 72 h postoperatively using a standard visual analog scale for evaluation of postepidural backache. A patient was considered to have postepidural backache when the postoperative visual analog scale score was higher than the preoperative score. RESULTS: The incidence of postepidural backache in patients in the control group for the 3 days were 22.8%, 17.4%, and 9.2%, all of which were significantly more frequent than observed in the patients in the tenoxicam group (6.8%, 4.0%, and 1.2%, P < 0.01). There was a significant association between backache and multiple attempts at epidural needle insertion. CONCLUSION: In summary, the local addition of tenoxicam reduced the incidence and severity of postepidural backache. PMID- 9856717 TI - Population pharmacokinetics of midazolam administered by target controlled infusion for sedation following coronary artery bypass grafting. AB - BACKGROUND: Midazolam is commonly used for short-term postoperative sedation of patients undergoing cardiac surgery. The purpose of this multicenter study was to characterize the pharmacokinetics and intersubject variability of midazolam in patients undergoing coronary artery bypass grafting. METHODS: With institutional review board approval, 90 consenting patients undergoing coronary artery bypass grafting were enrolled at three study centers. All subjects received sufentanil and midazolam via target-controlled infusions. After operation, midazolam was titrated to maintain deep sedation for at least 2 h. It was then titrated downward to decrease sedation for a minimum of 4 h more and was discontinued before tracheal extubation. Arterial blood samples were taken throughout the study and were assayed for midazolam and 1-hydroxymidazolam. Midazolam population pharmacokinetic parameters were estimated using NONMEM. Cross-validation was used to estimate the performance of the model. RESULTS: The pharmacokinetics of midazolam were best described by a simple three-compartment mammillary model. Typical pharmacokinetic parameters were V1 = 32.2 l, V2 = 53 l, V3 = 245 l, Cl1 = 0.43 l/min, Cl2 = 0.56 l/min, and Cl3 = 0.39 l/min. The calculated elimination half-life was 15 h. The median absolute prediction error was 25%, with a bias of 1.4%. The performance in the cross-validation was similar. Midazolam metabolites were clinically insignificant in all patients. CONCLUSIONS: The intersubject variability and predictability of the three-compartment pharmacokinetic model are similar to those of other intravenous anesthetic drugs. This multicenter study did not confirm previous studies of exceptionally large variability of midazolam pharmacokinetics when used for sedation in intensive care settings. PMID- 9856718 TI - Population pharmacodynamics of midazolam administered by target controlled infusion in SICU patients after CABG surgery. AB - BACKGROUND: Midazolam is used commonly for sedation in the surgical intensive care unit. A suboptimal dosing regimen may lead to relative overdosing, which could result in delayed extubation and increased cost. This multicenter trial characterized midazolam pharmacodynamics in patients recovering from coronary artery bypass grafting. METHODS: Three centers enrolled 90 patients undergoing coronary artery bypass grafting. All patients received sufentanil and midazolam via target-controlled infusion. After surgery, midazolam was titrated to a Ramsay sedation score of 5 for 2 h and then decreased to maintain a sedation score of 3 or 4 for at least another 4 h. Pharmacodynamic parameters were derived using NONMEM. The model was cross-validated to test performance. RESULTS: The probability of a given level of sedation was related to the midazolam concentration by this equation: P(Sedation > or = ss) = Cn/(Cn + C(50,ss)n), where ss is the sedation score, C is the sum of the midazolam concentration and a term reflecting the dissipating effect of anesthesia: C = [midazolam] + theta x e(-Kt), where theta = 256 ng/ml and K = 0.19 h(-1). C(50,ss) values for Ramsay scores of 2 to 6 were 5.7, 71, 171, 260, and 659 ng/ml, respectively. The model predicted 57% of the data points correctly and 88% within one sedation score. CONCLUSIONS: Despite previous reports of high interindividual variability in midazolam pharmacodynamics in patients in the surgical intensive care unit, these cross-validation results suggest that, when midazolam is administered using a target-controlled infusion device, the level of sedation can be predicted within 1 sedation score in 88% of patients based on the target midazolam concentration and the time since the conclusion of the anesthetic. PMID- 9856719 TI - Isobolographic analysis of the interactions between midazolam and propofol at GABA(A) receptors in embryonic mouse neurons. AB - BACKGROUND: Clinical studies suggest that midazolam and propofol interact synergistically to induce hypnosis, but these drugs do not interact synergistically to prevent movement in response to noxious stimuli. The mechanisms underlying these interactions are not certain but may occur at the level of the gamma-aminobutyric acid A (GABA(A)) receptor. METHODS: The authors evaluated the interactions between propofol and midazolam in modulating GABA(A) receptor activity in embryonic hippocampal neurons. The effects of midazolam and propofol on peak current evoked by submaximal concentrations of GABA were studied using the patch clamp method. Isobolographic analysis was undertaken by constructing concentration-response curves for midazolam and propofol alone and then evaluating the potency of combinations of midazolam and propofol. In other experiments, the concentration of GABA was increased and flurazepam was substituted for midazolam. RESULTS: Isobolographic analysis confirmed that midazolam and propofol interact synergistically to enhance currents evoked by low concentrations of GABA (1 microM). However, when the concentration of GABA was increased to 3 microM, the interaction was additive. The interaction between flurazepam and propofol was also additive for enhancement of currents evoked by 3 microM GABA. CONCLUSIONS: The interaction between midazolam and propofol was critically dependent on the concentration of GABA: Synergism was evident at low concentrations of GABA, but an additive interaction was apparent when the concentration of GABA was increased. Changes in GABA(A) receptor function may underlie the synergistic interaction between propofol and midazolam for clinical effects such as hypnosis. The clinical implication of the results is that the benefits of synergism observed at one concentration ratio of these drugs may not be apparent at another. PMID- 9856720 TI - Interaction of intrathecally infused morphine and lidocaine in rats (part I): synergistic antinociceptive effects. AB - BACKGROUND: Synergistic antinociception of opioids and local anesthetics has been established in bolus injections but not in long-term use. The somatic and visceral antinociceptive effects of intrathecally infused morphine or lidocaine were characterized, and the nature of the interaction of those agents in rats was evaluated. METHODS: Intrathecal catheters were implanted in rats. Morphine (0.3 to 10 microg x kg(-1) x h(-1)), lidocaine (30-1,000 microg x kg(-1) x h(-1)), a combination of those, or saline was infused intrathecally at a constant rate of 1 microl/h for 6 days. The tail flick and colorectal distension tests were used to measure the somatic and visceral antinociceptive effects, respectively. Nociceptive tests and motor function tests were repeated on days 1, 2, 3, 4, and 6. Isobolographic analysis was performed on the results of the tail flick test to determine the magnitude of the interaction. RESULTS: Intrathecally infused morphine produced dose-dependent antinociceptive effects in both the tail flick and the colorectal distension tests. Morphine showed a lower peak percentage maximum possible effect (%MPE) in the colorectal distension test than in the tail flick test. Intrathecal lidocaine also produced dose-dependent antinociceptive effects. Lidocaine infusion at 1,000 microg x kg(-1) x h(-1) caused motor impairment. Coinfusion of morphine 0.3 microg x kg(-1) x h(-1) and lidocaine 200 microg x kg(-1) x h(-1), which had no effects by themselves, significantly increased the percentage maximum possible effects (P < 0.01). Coinfused lidocaine potentiated the duration and the magnitude of morphine antinociception. Isobolographic analysis of the tail flick test on day 1 showed a synergistic interaction between morphine and lidocaine. CONCLUSIONS: Morphine and lidocaine intrathecally coadministered synergistically potentiated the antinociceptive effects of each other. That coinfusion dramatically potentiated visceral antinociception, whereas the infusion of morphine alone showed little visceral antinociception. PMID- 9856721 TI - Interaction of intrathecally infused morphine and lidocaine in rats (part II): effects on the development of tolerance to morphine. AB - BACKGROUND: There has been little information regarding the effects of local anesthetics on tolerance to opioids, although chronic use of combination of opioids and local anesthetics is popular for pain control. This study was designed to examine the effects of lidocaine on morphine tolerance to somatic and visceral antinociception. METHODS: Rats received a continuous intrathecal infusion of morphine (0.3-10 microg x kg(-1) x h(-1)), lidocaine (30-1000 microg x kg(-1). h(-1)), a combination of those, or saline. After 6- day infusion, intrathecal morphine challenge test (5 microg/10 microl) was performed, and time response curve was constructed to assess the magnitude of tolerance. The tail flick (TF) test and colorectal distension (CD) test were used to measure somatic and visceral antinociceptive effects, respectively. RESULTS: Antinociceptive effects in the TF and CD tests caused by morphine challenge were reduced (P < 0.01) in the morphine infused groups. The magnitude of the tolerance was inversely associated with the amount of morphine infused. Lidocaine infusion induced no different change in the morphine challenge test from that seen in the saline infusion group. Development of tolerance was greater in morphine 3 microg x kg(-1) h(-1) than in morphine 0.75 microg x kg(-1) x h(-1) + lidocaine 150 microg x kg(-1) x h(-1) despite their similar antinociceptive effects during intrathecal infusion. The infusion of a low dose of morphine (0.3 microg kg(-1) x h(-1)) did not reduce the antinociceptive effects in the challenge test. CONCLUSION: Lidocaine in combination with morphine does not reduce tolerance to morphine nor develop cross-tolerance. The intrathecal infusion of morphine induced tolerance to somatic and visceral antinociception in a dose-dependent fashion. PMID- 9856722 TI - Carotid body chemoreceptor function is impaired by vecuronium during hypoxia. AB - BACKGROUND: Neuromuscular blocking agents reduce the human ventilatory response to hypoxia at partial neuromuscular block. It was hypothesized that vecuronium impairs carotid body chemoreceptor function during hypoxia. METHOD: The effect of systemic administration of vecuronium on single chemoreceptor activity during hypoxia, as recorded from a single nerve fiber preparation of the carotid sinus nerve, was studied in seven mechanically ventilated New Zealand White rabbits during continuous thiopental anesthesia. During normoventilation, the isocapnic hypoxic chemosensitivity of the single carotid body chemoreceptor was measured at four levels of oxygenation; these measurements were repeated at six separate occasions: control recording before injection, after intravenous administrations of 0.1 mg and 0.5 mg of vecuronium, and then at three occasions during a 90-min recovery period. Chemoreceptor chemosensitivity during isocapnic hypoxia was expressed as a hyperbolic function: Chemoreceptor output (Hz) = a + b x PaO2(-1) (mmHg). RESULTS: Chemosensitivity was reduced after both 0.1 mg and 0.5 mg vecuronium intravenous administration compared with control measurements; the hypoxic response curve was significantly depressed after both doses (P < 0.05). Notably, there was variation in the effect of vecuronium; some chemoreceptor preparations showed only minimal impairment, whereas some showed an almost abolished response to hypoxia. The chemosensitivity remained significantly depressed at 30 and 60 min but had recovered spontaneously at 90 min after 0.5 mg vecuronium. DISCUSSION: It is concluded that vecuronium depresses carotid body chemoreceptor function to a varying extent during hypoxia and that the depression recovers spontaneously. PMID- 9856723 TI - Local cerebral blood flow, local cerebral glucose utilization, and flow metabolism coupling during sevoflurane versus isoflurane anesthesia in rats. AB - BACKGROUND: Compared to isoflurane, knowledge of local cerebral glucose utilization (LCGU) and local cerebral blood flow (LCBF) during sevoflurane anesthesia is limited. METHODS: LCGU, LCBF, and their overall means were measured in Sprague-Dawley rats (8 groups, n=6 each) during sevoflurane and isoflurane anesthesia, 1 and 2 MAC, and in conscious control animals (2 groups, n=6 each) using the autoradiographic 2-[14C]deoxy-D-glucose and 4-iodo-N-methyl [14C]antipyrine methods. RESULTS: During anesthesia, mean cerebral glucose utilization was decreased: control, 56+/-5 micronmol x 100 g(-1) x min(-1); 1 MAC isoflurane, 32+/-4 micromol x 100 g(-1) x min(-1) (-43%); 1 MAC sevoflurane, 37+/ 5 micromol x 100 g(-1) x min(-1) (-34%); 2 MAC isoflurane, 23+/-3 micromol x 100 g(-1) x min(-1) (-58%); 2 MAC sevoflurane, 23+/-5 micromol x 100 g(-1) x min(-1) (-59%). Local analysis showed a reduction in LCGU in the majority of the 40 brain regions analyzed. Mean cerebral blood flow was increased as follows: control 93+/ 8 ml x 100 g(-1) x min(-1); 1 MAC isofurane, 119+/-19 ml x 100 g(-1) x min(-1) (+28%); 1 MAC sevoflurane, 104+/-15 ml x 100 g(-1) x min(-1) (+12%); 2 MAC isoflurane, 149+/-17 ml x 100 g(-1) x min(-1) (+60%); 2 MAC sevoflurane, 118+/-21 ml x 100 g(-1) min(-1) (+27%). LCBF was increased in most brain structures investigated. Correlation coefficients obtained for the relationship between LCGU and LCBF were as follows: control 0.93; 1 MAC isoflurane, 0.89; 2 MAC isoflurane, 0.71; 1 MAC sevoflurane, 0.83; 2 MAC sevoflurane, 0.59). CONCLUSION: Mean and local cerebral blood flows were lower during sevoflurane than during isoflurane anesthesia. This difference cannot be explained by differing changes in glucose utilization because glucose utilization was decreased to the same extent in both groups. PMID- 9856724 TI - Aspirin synergistically potentiates isoflurane minimum alveolar concentration reduction produced by morphine in the rat. AB - BACKGROUND: The combination of opioids and nonsteroidal antiinflammatory drugs is more analgesic than the summed effect of each drug administered separately. This synergism has been used to obtain analgesia in the postoperative period at doses at which side effects are minimal. The aim of this study is to evaluate the analgesic interaction between aspirin and morphine in the rat during isoflurane anesthesia. The reduction in minimum alveolar concentration of isoflurane (MAC(ISO)) was used as an objective measure of the analgesic potency of individual drugs and their use in combination. METHODS: Thirty-seven male Wistar rats were anesthetized with isoflurane in oxygen, and the MAC(ISO) was determined before and after the intravenous administration of aspirin and morphine. Rats were administered morphine alone (1, 3, and 10 mg/kg) or morphine (1 and 3 mg/kg) and aspirin (30 mg/kg). The MAC(ISO) was determined from alveolar gas samples at the time of tail clamp. The duration of MAC(ISO) reduction was recorded. RESULTS: Aspirin did not have an effect on MAC(ISO), (average, 1.35+/-0.1%), whereas the combination of morphine (1 and 3 mg/kg) and aspirin (30 mg/kg) produced a reduction in the dose of morphine needed to produce the same degree of MAC(ISO) reduction. Actual MAC(ISO+drug) data were as follows: 1 mg/kg morphine, 1.17+/ 0.14%; 3 mg/kg morphine, 0.98+/-0.15%; 1 mg/kg morphine plus aspirin, 0.90+/ 0.04%; 10 mg/kg morphine, 0.63+/-0.13%; and 3 mg/kg morphine plus aspirin, 0.64+/ 0.06%. CONCLUSIONS: The synergistic effects of aspirin and morphine allow a clinically significant reduction in the requirements of isoflurane and isoflurane plus morphine, and these drug combinations may decrease the side effects associated with the use of single higher, equianalgesic doses of these drugs. PMID- 9856725 TI - Propofol-induced increase in vascular capacitance is due to inhibition of sympathetic vasoconstrictive activity. AB - BACKGROUND: Venodilation is thought to be one of the mechanisms underlying propofol-induced hypotension. The purpose of this study is to test two hypotheses: (1) propofol increases systemic vascular capacitance, and (2) the capacitance change produced by propofol is a result of an inhibition of sympathetic vasoconstrictor activity. METHODS: In 33 Wistar rats previously anesthetized with urethane and ketamine, vascular capacitance was examined before and after propofol infusion by measuring mean circulatory filling pressure (Pmcf). The Pmcf was measured during a brief period of circulatory arrest produced by inflating an indwelling balloon in the right atrium. Rats were assigned into four groups: an intact group, a sympathetic nervous system (SNS) block group produced by hexamethonium infusion, a SNS-block + noradrenaline (NA) group, and a hypovolemic group. The Pmcf was measured at a control state and 2 min after a bolus administration of 2, 10, and 20 mg/kg of propofol. RESULTS: The mean arterial pressure (MAP) was decreased by propofol dose-dependently in intact, hypovolemic, and SNS-block groups, but the decrease in MAP was less in the SNS-block group (-25%) than in the intact (-50%) and hypovolemic (-61%) groups. In the SNS-block + NA group, MAP decreased only at 20 mg/kg of propofol ( 18%). The Pmcf decreased in intact and hypovolemic groups in a dose-dependent fashion but was unchanged in the SNS-block and SNS-block + NA groups. CONCLUSIONS: The results have provided two principal findings: (1) propofol decreases Pmcf dose-dependently, and (2) the decrease in Pmcf by propofol is elicited only when the sympathetic nervous system is intact, suggesting that propofol increases systemic vascular capacitance as a result of an inhibition of sympathetic nervous system. PMID- 9856727 TI - Common genetic determinants of halothane and isoflurane potencies in Caenorhabditis elegans. AB - BACKGROUND: Genetics provides a way to evaluate anesthetic action simultaneously at the molecular and behavioral levels. Results from strains that differ in anesthetic sensitivity have been mixed in their support of unitary theories of anesthesia. Here the authors use the previously demonstrated large variation of halothane sensitivities in Caenorhabditis elegans recombinant inbred strains to assess the similarities of the determinants of halothane action with those of another volatile anesthetic, isoflurane. METHODS: The recombinant inbred strains, constructed from two evolutionarily distinct C. elegans lineages, were phenotyped. A coordination assay on agar quantified the sensitivity to the volatile anesthetics; median effective concentrations (EC50s) were calculated by nonlinear regression of concentration-response data and were correlated between the drugs for those strains tested in common. Genetic loci were identified by statistical association between EC50s and chromosomal markers. RESULTS: The recombinant inbred strains varied dramatically in sensitivity to halothane and isoflurane, with a 10-fold range in EC50s. Heritability estimates for each drug were imprecise but altogether high (49-80%). Halothane and isoflurane EC50s were significantly correlated (r=0.71, P < 10(-9)). Genetic loci controlling sensitivity were found for both volatile anesthetics; the most significant determinant colocalized on chromosome V. A smaller recombinant inbred strain study of ethanol-induced immobility segregated different genetic effects that did not correlate with sensitivity to either halothane or isoflurane. CONCLUSIONS: The genetic determinants driving the large variation in anesthetic sensitivity in these C. elegans recombinant inbred strains are very similar for halothane and isoflurane sensitivity. PMID- 9856726 TI - Preservation of hypoxic pulmonary vasoconstriction during sevoflurane and desflurane anesthesia compared to the conscious state in chronically instrumented dogs. AB - BACKGROUND: The authors' objective was to assess the extent to which sevoflurane and desflurane anesthesia alter the magnitude of hypoxic pulmonary vasoconstriction compared with the response measured in the same animal in the conscious state. METHODS: Left pulmonary vascular pressure-flow plots were generated in seven chronically instrumented dogs by continuously measuring the pulmonary vascular pressure gradient (pulmonary arterial pressure-left atrial pressure) and left pulmonary blood flow during gradual (approximately 1 min) inflation of a hydraulic occluder implanted around the right main pulmonary artery. Pressure-flow plots were generated during normoxia and hypoxia on separate days in the conscious state, during sevoflurane (approximately 3.5% end tidal), and during desflurane (approximately 10.5% end-tidal) anesthesia. Values are mean+/-SEM. RESULTS: In the conscious state, administration of the hypoxic gas mixture by conical face mask decreased (P < 0.01) systemic arterial PO2 from 94+/-2 mmHg to 50+/-1 mmHg and caused a leftward shift (P < 0.01) in the pressure flow relationship, indicating pulmonary vasoconstriction. The magnitude of hypoxic pulmonary vasoconstriction in the conscious state was flow-dependent (P < 0.01). Neither anesthetic had an effect on the baseline pressure-flow relationship during normoxia. The magnitude of hypoxic pulmonary vasoconstriction during sevoflurane and desflurane was also flow-dependent (P < 0.01). Moreover, at any given value of flow the magnitude of hypoxic pulmonary vasoconstriction was similar during sevoflurane and desflurane compared with the conscious state. CONCLUSION: These results indicate that hypoxic pulmonary vasoconstriction is preserved during sevoflurane and desflurane anesthesia compared with the conscious state. Thus, inhibition of hypoxic pulmonary vasoconstriction is not a general characteristic of inhalational anesthetics. The flow-dependent nature of the response should be considered when assessing the effects of physiologic or pharmacologic interventions on the magnitude of hypoxic pulmonary vasoconstriction. PMID- 9856728 TI - Role of spinal NO in antiallodynic effect of intrathecal clonidine in neuropathic rats. AB - BACKGROUND: The role of spinal nitric oxide (NO) in neuropathic pain remains uncertain. Although intrathecal clonidine causes NO release in the spinal cord, the functional role of spinal NO in clonidine-produced analgesia has not been examined. The objectives of this study were to assess the role of spinal NO in maintenance of allodynia and to determine the role of spinal NO in the antiallodynic effect of intrathecal clonidine. METHODS: Allodynia was produced in rats by tight ligation of the left L5-L6 spinal nerves. Intrathecal catheters were inserted with tips in the lumbar intrathecal space. Mechanical allodynia was determined by application of von Frey filaments to the left hindpaw. In the first series of experiments, allodynia was assessed before and after intrathecal injection of saline, L-arginine, an NO donor (SNAP), two NO synthase inhibitors (TRIM and NMMA), or an NO scavenger (PTIO). In the second series of experiments, 20 microg of clonidine was injected intrathecally 15 min after intrathecal injection of saline, TRIM, NMMA, or PTIO. RESULTS: Allodynia was not affected significantly by intrathecal injection of L-arginine, SNAP, TRIM, NMMA, or PTIO. The antiallodynic effect produced by intrathecal injection of clonidine was attenuated significantly by pretreatment with TRIM, NMMA, or PTIO. CONCLUSIONS: These results demonstrate that spinal NO neither contributes significantly to maintenance of allodynia nor produces detectable antiallodynic effect in this neuropathic pain model. Furthermore, this study provides functional evidence that spinal NO plays an important role in the antiallodynic effect of intrathecal clonidine in neuropathic pain. PMID- 9856729 TI - Recovery from sevoflurane anesthesia: a comparison to isoflurane and propofol anesthesia. AB - BACKGROUND: Sevoflurane has a lower blood:gas partition coefficient than isoflurane, which may cause a more rapid recovery from anesthesia; it also might cause faster emergence times than for propofol-based anesthesia. We evaluated a database that included recovery endpoints from controlled, randomized, prospective studies sponsored by Abbott Laboratories that compared sevoflurane to isoflurane or propofol when extubation was planned immediately after completion of elective surgery in adult patients. METHODS: Sevoflurane was compared to isoflurane in eight studies (N=2,008) and to propofol in three studies (N=436). Analysis of variance was applied using least squares method mean values to calculate the pooled mean difference in recovery endpoints between primary anesthetics. The effects of patient age and case duration also were determined. RESULTS: Sevoflurane resulted in statistically significant shorter times to emergence (-3.3 min), response to command (-3.1 min), orientation (-4.0 min) and first analgesic (-8.9 min) but not time to eligibility for discharge (-1.7 min) compared to isoflurane (mean difference). Times to recovery endpoints increased with increasing case duration with isoflurane but not with sevoflurane (patients receiving isoflurane took 4-5 min more to emerge and respond to commands and 8.6 min more to achieve orientation during cases longer than 3 hr in duration than those receiving sevoflurane). Patients older than 65 yr had longer times to orientation, but within any age group, orientation was always faster after sevoflurane. There were no differences in recovery times between sevoflurane and propofol. CONCLUSIONS: Recovery from sevoflurane was 3-4 min faster than with isoflurane in all age groups, and the difference was magnified in longer-duration surgical cases (> 3 hr). PMID- 9856730 TI - Differential effects of fentanyl and morphine on intracellular Ca2+ transients and contraction in rat ventricular myocytes. AB - BACKGROUND: Our objective was to elucidate the direct effects of fentanyl and morphine on cardiac excitation-contraction coupling using individual, field stimulated rat ventricular myocytes. METHODS: Freshly isolated myocytes were loaded with fura-2 and field stimulated (0.3 Hz) at 28 degrees C. Amplitude and timing of intracellular Ca2+ concentration (at a 340:380 ratio) and myocyte shortening (video edge detection) were monitored simultaneously in individual cells. Real time Ca2+ uptake into isolated sarcoplasmic reticulum vesicles was measured using fura-2 free acid in the extravesicular compartment. RESULTS: The authors studied 120 cells from 30 rat hearts. Fentanyl (30-1,000 nM) caused dose dependent decreases in peak intracellular Ca2+ concentration and shortening, whereas morphine (3-100 microM) decreased shortening without a concomitant decrease in the Ca2+ transient. Fentanyl prolonged the time to peak and to 50% recovery for shortening and the Ca2+ transient, whereas morphine only prolonged the timing parameters for shortening. Morphine (100 microM), but not fentanyl (1 microM), decreased the amount of Ca2+ released from intracellular stores in response to caffeine in intact cells, and it inhibited the rate of Ca2+ uptake in isolated sarcoplasmic reticulum vesicles. Fentanyl and morphine both caused a downward shift in the dose-response curve to extracellular Ca2+ for shortening, with no concomitant effect on the Ca2+ transient. CONCLUSIONS: Fentanyl and morphine directly depress cardiac excitation-contraction coupling at the cellular level. Fentanyl depresses myocardial contractility by decreasing the availability of intracellular Ca2+ and myofilament Ca2+ sensitivity. In contrast, morphine depresses myocardial contractility primarily by decreasing myofilament Ca2+ sensitivity. PMID- 9856731 TI - Halothane attenuates calcium sensitization in airway smooth muscle by inhibiting G-proteins. AB - BACKGROUND: Halothane directly relaxes airway smooth muscle partly by decreasing the Ca2+ sensitivity. In smooth muscle, receptor stimulation is thought to increase Ca2+ sensitivity via a cascade of heterotrimeric and small monomeric guanine nucleotide-binding proteins (G-proteins). Whether this model is applicable in the airway and where halothane acts in this pathway were investigated. METHODS: A beta-escin-permeabilized canine tracheal smooth muscle preparation was used. Exoenzyme C3 of Clostridium botulinum, which inactivates Rho monomeric G-proteins, was used to evaluate the involvement of this protein in the Ca2+ sensitization pathway. The effects of halothane on different stimulants acting at different levels of signal transduction were compared: acetylcholine on the muscarinic receptor, aluminum fluoride (AIF4-) on heterotrimeric G-proteins, and guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) on all G-proteins. RESULTS: Exoenzyme C3 equally attenuated acetylcholine- and AIF4--induced Ca2+ sensitization, suggesting that these pathways are both mediated by Rho. Halothane applied before stimulation equally attenuated acetylcholine- and AIF4--induced Ca2+ sensitization. However, when added after Ca2+ sensitization was established, the effect of halothane was greater during Ca2+ sensitization induced by acetylcholine compared with AIF4-, which, along with the previous result, suggests that halothane may interfere with dissociation of heterotrimeric G proteins. Halothane applied during GTPgammaS-induced Ca2+ sensitization had no significant effect on force, suggesting that halothane has no effect downstream from monomeric G-proteins. CONCLUSION: Halothane inhibits increases in Ca2+ sensitivity of canine tracheal smooth muscle primarily by interfering with the activation of heterotrimeric G-proteins, probably by inhibiting their dissociation. PMID- 9856732 TI - Assessment of teratogenic effects of lidocaine in rat embryos cultured in vitro. AB - BACKGROUND: Lidocaine has been reported to cause neural tube closure defects in vitro in mice at clinically relevant concentrations. However, no studies have been conducted to further investigate this potentially hazardous effect of lidocaine. This study was aimed to reassess teratogenic effects of lidocaine in vitro in rats. METHODS: Sprague-Dawley rat embryos were explanted at 8:00 AM on gestational day 9 and were cultured in medium containing various concentrations of lidocaine. (Embryos in the control group were cultured without lidocaine). After 50 h of culture, they were evaluated for growth size and morphology, including the neural tube closure. RESULTS: In the presence of 250 microM of lidocaine, embryos showed a increased incidence of situs inversus compared with control group but were otherwise normal. At 375 microM, embryos showed slight growth retardation but no significant morphologic abnormalities. At 500 microM, all viable embryos showed severe morphologic abnormalities. However, morphologic abnormalities were so-called nonspecific types and neural tube closure defects were not observed. CONCLUSIONS: Results from the current study indicate that lidocaine causes teratogenic effects in vitro in rats only at concentrations much higher than clinically relevant concentrations. Furthermore, lidocaine did not cause neural tube closure defects at any concentrations evaluated. PMID- 9856733 TI - Obstetric postanesthesia care unit stays: reevaluation of discharge criteria after regional anesthesia. AB - BACKGROUND: Obstetric patients may have long postanesthesia care unit (OB-PACU) stays after surgery because of residual regional block or other conditions. This study evaluated whether modified discharge criteria might allow for earlier discharge without compromising patient safety. METHODS: Data were prospectively collected for 6 months for all patients (N=358) who underwent cesarean section or tubal ligation and recovered in the OB-PACU. Regional anesthesia was used in 94% of patients. The duration of anesthesia and PACU stays, the presence and treatment of events in the PACU, and the regression of neural blockade were recorded. Discharge from the OB-PACU required a 60-min minimum stay, stable vital signs, adequate analgesia, and ability to flex the knees. After completion of prospective data collection, events that kept patients in the PACU after 60 min were reevaluated as to whether patients needed to stay in the PACU for medical reasons. "Needed to stay" events included bleeding, cardiorespiratory problems, sedation, dizziness, and pain. "Safe to leave" conditions included pruritus, nausea, and residual neural blockade. The cumulative duration of OB-PACU stays not clearly justifiable for medical reasons was calculated. RESULTS: Residual block and spinal opioid side effects accounted for the majority of "unnecessary" stays. Annually, 429 h of PACU time could have been saved using the revised criteria. Complications did not develop subsequently in any patient deemed "safe to leave." CONCLUSIONS: In many obstetric patients, the duration of PACU stays could safely be shortened by continuing observation in a lower-acuity setting. This may result in greater flexibility and more efficient use of nursing personnel. PMID- 9856734 TI - Intraoperative heparin flushes and subsequent acute heparin-induced thrombocytopenia. PMID- 9856735 TI - Effects of sevoflurane on QT interval in a patient with congenital long QT syndrome. PMID- 9856736 TI - The EXIT procedure facilitates delivery of an infant with a pretracheal teratoma. PMID- 9856737 TI - Airway fire during tracheostomy: extubation may be contraindicated. PMID- 9856738 TI - Streptococcus salivarius meningitis after spinal anesthesia. PMID- 9856739 TI - Pituitary apoplexy following mitral valve repair. PMID- 9856740 TI - Early hemodynamic changes following emergency mitral valve replacement for traumatic mitral insufficiency following balloon mitral valvotomy: report of six cases. PMID- 9856741 TI - Improvement of extreme hypoxemia during end-stage congenital heart disease using nasal nitric oxide. PMID- 9856742 TI - An unexpected arousal effect of etomidate in a patient on high-dose steroids. PMID- 9856743 TI - Fatal pulmonary fat embolism in the early postoperative period. PMID- 9856744 TI - Continuous cardiac output catheters: delay in in vitro response time after controlled flow changes. PMID- 9856745 TI - Foreign body in the trachea originated from the inner wall of the Univent tube. PMID- 9856746 TI - Rural realities. PMID- 9856748 TI - Perfuse or precondition? PMID- 9856747 TI - The blood sparing effect of aprotinin should be revisited. PMID- 9856749 TI - No evidence of ischemic preconditioning during coronary revascularization. PMID- 9856750 TI - Value of presenting the time-course of pain relief in analgesic trials. PMID- 9856751 TI - Pulse oximetry probe adhesive disks: a potential for infant aspiration. PMID- 9856752 TI - Complication of Bullard laryngoscope: dislodgment of blade-extender resulting in an upper airway foreign body. PMID- 9856753 TI - High-concentration cisatracurium in intensive care unit solution. PMID- 9856754 TI - Bowing of the needle technique for nerve blocks. PMID- 9856756 TI - A rare cause of fire in the operating room. PMID- 9856755 TI - Unreasonably low bispectral index values in a volunteer with genetically determined low-voltage electroencephalographic signal. PMID- 9856757 TI - Incompatibility of propofol emulsion with anesthetic drugs. PMID- 9856758 TI - Photoreceptor membrane proteins, phototransduction, and retinal degenerative diseases. The Friedenwald Lecture. PMID- 9856759 TI - New discoveries on the roles of matrix metalloproteinases in ocular cell biology and pathology. PMID- 9856760 TI - Advanced glycation end products in vitreous: Structural and functional implications for diabetic vitreopathy. AB - PURPOSE: Advanced glycation end products (AGEs) form irreversible cross-links with many macromolecules and have been shown to accumulate in tissues at an accelerated rate in diabetes. In the present study, AGE formation in vitreous was examined in patients of various ages and in patients with diabetes. Ex vivo investigations were performed on bovine vitreous incubated in glucose to determine AGE formation and cross-linking of vitreous collagen. METHODS: By means of an AGE-specific enzyme-linked immunosorbent assay (ELISA), AGE formation was investigated in vitreous samples obtained after pars plana vitrectomy in patients with and without diabetes. In addition, vitreous AGEs were investigated in bovine vitreous collagen after incubation in high glucose, high glucose with aminoguanidine, or normal saline for as long as 8 weeks. AGEs and AGE cross linking was subsequently determined by quantitative and qualitative assays. RESULTS: There was a significant correlation between AGEs and increasing age in patients without diabetes (r = 0.74). Furthermore, a comparison between age matched diabetic and nondiabetic vitreous showed a significantly higher level of AGEs in the patients with diabetes (P < 0.005). Collagen purified from bovine vitreous incubated in 0.5 M glucose showed an increase in AGE formation when observed in dot blot analysis, immunogold labeling, and AGE ELISA. Furthermore, there was increased cross-linking of collagen in the glucose-incubated vitreous, when observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and protein separation. This cross-linking was effectively inhibited by coincubation with 10 mM aminoguanidine. CONCLUSIONS: This study suggests that AGEs may form in vitreous with increasing age. This process seems to be accelerated in the presence of diabetes and as a consequence of exposure to high glucose. Advanced glycation and AGE cross-linking of the vitreous collagen network may help to explain the vitreous abnormalities characteristic of diabetes. PMID- 9856761 TI - Missense mutations in the PAX6 gene in aniridia. AB - PURPOSE: Aniridia is caused by a mutation of the PAX6 gene. Haploinsufficiency of the gene product is thought to result in the aniridia phenotype, because most mutations thus far detected have been large deletions encompassing the entire gene and nonsense, frameshift, or splice errors that result in premature translational termination on one of the alleles. Only two missense mutations have been detected in aniridia pedigrees, each of which occurs in its paired domain or homeodomain. In this study, four novel missense mutations were found in three aniridia pedigrees. METHODS: Polymerase chain reaction-single-strand conformation polymorphism analysis and sequencing of the PAX6 gene were performed using genomic DNA of three aniridia pedigrees and more than 100 healthy control subjects. RESULTS: Three mutations occurred in the N-terminal subdomain of the paired domain, namely N17S, I29V, and R44Q, the first two of which were detected on the same allele of one patient. The other mutation (Q178H) was in the linking portion of the paired domain and homeodomain. CONCLUSIONS: These missense mutations give rise to haploinsufficiency by another route, because the missense mutations presented here resulted in an aniridia phenotype indistinguishable from that caused by a heterozygous deletion of the entire PAX6 gene. PMID- 9856762 TI - Selective degradation of nonsense beta-phosphodiesterase mRNA in the heterozygous rd mouse. AB - PURPOSE: To investigate the molecular mechanism relating phenotype and genotype in the rd mouse, mRNA and pre-mRNA levels derived from the wild-type and position 347 nonsense mutant beta-phosphodiesterase (beta-PDE) genes were determined and compared with the corresponding gene copy ratios. METHODS: Total RNA and genomic DNA was isolated from the retinas of three heterozygous rd/+ mouse strains. For each, quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the ratio of wild-type and rd beta-phosphodiesterase pre-mRNA and mature mRNA. The gene copy ratio between wild-type and rd beta-PDE was also determined by quantitative PCR. RESULTS: The pre-mRNA ratio of wild-type versus nonsense mutant was close to 1:1, whereas the corresponding mRNA ratio was greater than 3:1, even though the gene copy ratio was confirmed to be 1:1. CONCLUSIONS: The equivalence of pre-mRNA ratio level for wild-type and nonsense mutant in the rd/+ retina indicates that both genes were transcribed at similar levels. Thus, neither the nonsense mutation at position 347 nor the intron 1 retroviral insertion also present in the rd gene seem to have affected gene transcription. In contrast, the strain-independent bias favoring wild-type mature mRNA in vivo suggests a specific degradation of mutant transcript during or after pre-mRNA splicing. This allele-specific degradation serves to decrease mutant transcript levels dramatically in all rd strains, and suggests that photoreceptor cells have the capacity to reduce the level of an mRNA containing a nonsense mutation. PMID- 9856763 TI - Baseline findings in the Collaborative Longitudinal Evaluation of Keratoconus (CLEK) Study. AB - PURPOSE: To describe the baseline findings in patients enrolled in the Collaborative Longitudinal Evaluation of Keratoconus (CLEK) Study. METHODS: This is a longitudinal observational study of 1209 patients with keratoconus enrolled at 16 clinical centers. Its main outcome measures are corneal scarring, visual acuity, keratometry, and quality of life. RESULTS: The CLEK Study patients had a mean age of 39.29+/-10.90 years with moderate to severe disease, assessed by a keratometric-based criterion (95.4% of patients had steep keratometric readings of at least 45 D) and relatively good visual acuity (77.9% had best corrected visual acuity of at least 20/40 in both eyes). Sixty-five percent of the patients wore rigid gas-permeable contact lens, and most of those (73%) reported that their lenses were comfortable. Only 13.5% of patients reported a family history of keratoconus. None reported serious systemic diseases that had been previously reported to be associated with keratoconus. Many (53%) reported a history of atopy. Fifty-three percent had corneal scarring in one or both eyes. CONCLUSIONS: Baseline findings suggest that keratoconus is not associated with increased risk of connective tissue disease and that most patients in the CLEK Study sample represent mild to moderate keratoconus. Additional follow-up of at least 3 years will provide new information about the progression of keratoconus, identify factors associated with progression, and assess its impact on quality of life. PMID- 9856764 TI - SPARC from corneal epithelial cells modulates collagen contraction by keratocytes. AB - PURPOSE: Contraction of the scar tissue during corneal wound healing changes the shape of the cornea and corneal refraction. In a previous study, it was found that corneal epithelial cells secrete the factor that stimulates collagen gel contraction by keratocytes in vitro. The purpose of the present study was to purify and identify the contraction-stimulating factor derived from corneal epithelial cells. METHODS: The cultured medium of rabbit corneal epithelial cells was collected and used as an epithelial cell-conditioned medium (ECCM). Subcultured rabbit keratocytes were embedded in a collagen gel, and collagen gel contraction was investigated. The contraction-stimulating factor in the ECCM was purified through acetone precipitation, affinity chromatography (heparin Sepharose), gel filtration, and reversed-phase chromatography. The amino acid sequence of a contraction-stimulating factor was analyzed. RESULTS: Collagen gel contraction by keratocytes was enhanced by the addition of ECCM in a dose dependent manner. The amino acids sequence of the contraction-stimulating factor was homologous to a 32-kDa glycoprotein, a secreted protein that is acidic and rich in cysteine (SPARC). Western blot analysis confirmed that SPARC was contained in the ECCM. Collagen gel contraction by keratocytes was enhanced by the addition of purified SPARC in a dose-dependent manner. SPARC was found in the basal layer of the migrating epithelium and activated keratocytes adjacent to the wound 3 days and 1 week after perforating injury in rabbit corneas. CONCLUSIONS: Epithelial cells secrete SPARC, which modulates the contraction of scar tissue in the corneal stroma. PMID- 9856765 TI - Identification of the cornea-specific keratin 12 promoter by in vivo particle mediated gene transfer. AB - PURPOSE: Keratin 12 (K12) is a cornea epithelial cell-specific intermediate filament component. To provide a better understanding of its expression, it is necessary to identify and characterize the promoter of Krt1.12 gene. METHODS: The 2.5-kb DNA 5' to Krt1.12 gene was sequenced. Krt1.12 promoter-beta-gal DNA constructs were prepared and used in vivo to transfect rabbit corneas, conjunctivas, and skin by particle-mediated gene transfer (Gene Gun). In vitro, the DNA constructs were transfected into cultured T-antigen-transformed rabbit corneal epithelial (RCE-T) cells and human fibrosarcoma HT-1080 fibroblasts with lipofectamine. The promoter activity was assessed by measuring beta-gal (beta galactosidase) activity using histochemical staining with 5-Bromo-4-chloro-3 indolyl-beta-D-galactoside and enzyme assay with o-nitrophenyl beta-D galactopyranoside. RESULTS: There are four Pax-6 pair box binding elements found between -910 and -2000 bp 5'-flanking the transcription initiation site of the Krt1.12 gene. None of promoter constricts can be expressed by HT-1080 cells. Cotransfection of Pax-6 cDNA with K12 promoter-beta-gal constructs containing Pax 6 elements results in a fourfold increase of beta-gal activities in RCE-T cells but not HT-1080 fibroblasts. The data of in vivo transfection in the rabbit by Gene Gun indicate that reporter gene constructs containing 0.6-kb and longer DNA fragments 5'-flanking Krt1.12 gene are effectively expressed in corneal, but not conjunctival or epidermal epithelial cells. CONCLUSIONS: The particle-mediated gene transfer is a suitable technique for in vivo delivery of transgenes to corneal epithelial cells. The 2.5-kb DNA fragment 5'-flanking Krt1.12 contains corneal epithelial cell-specific regulatory cis-DNA elements. Pax-6 is a positive transcription factor essential for keratin 12 expression. PMID- 9856766 TI - Human corneal epithelial cell functional responses to inflammatory agents and their antagonists. AB - PURPOSE: To investigate the epithelial nature of primary and SV40 virus immortalized human corneal epithelial (CEPI) cells and to study a variety of functional responses to some key inflammatory agents (bradykinin [BK], histamine, and platelet-activating factor [PAF]) and their antagonists in these cells. METHODS: Primary CEPI (P-CEPI) and clone 4 of the SV40 virus-immortalized (CEPI 17-CL4) cells were analyzed for their interaction with several monoclonal antibodies selective for various cytokeratins to define their immunocytochemical characteristics and phenotypic traits. Both cell types were tested for their ability to respond to BK, histamine, and PAF and their antagonists, using the production of [3H]inositol phosphates ([3H]IPs) as an index of receptor activation. The ability of BK, PAF, and histamine to stimulate cytokine release and the induction of mRNA for matrix metalloproteinase-1 (MMP-1) were also studied using enzyme-linked immunosorbent assay and reverse transcriptase polymerase chain reaction techniques, respectively. RESULTS: P-CEPI and CEPI-17 CL4 cells were both shown to possess the epithelial cell cytokeratins labeled with AE1 and AE3 antibodies. The potencies (EC50s) of BK, histamine, and PAF were similar for stimulating [3H]IPs production in P-CEPI and CEPI-17-CL4 cells: BK = 2.27 to 2.99 nM, PAF = 17.1 to 18.26 nM, and histamine = 1.65 to 5.74 microM (all n = 3 to 6). Both cell types also responded similarly to receptor-selective antagonists for BK, PAF, and histamine (Hoe-140: Ki = 10.1 to 11.9 nM; PCA-4248: Ki = 315 to 421 nM; triprolidine: Ki = 0.8 to 4.76 nM; all n = 5 to 10). Histamine (100 microM) and interleukin-1alpha (IL-1alpha, 10 ng/ml) significantly stimulated IL-6 and granulocyte macrophage colony-stimulating factor release, and histamine, BK, and PAF stimulated the mRNA for MMP-1 in these cells. CONCLUSIONS: These studies have shown that the primary and immortalized human corneal epithelial cells express functional BK (a B2 subtype), histamine (an H1 subtype), and PAF receptors and exhibit very similar immunocytochemical, signal transduction, and pharmacological properties. Therefore, the CEPI-17-CL4 cells (currently at passage 220) appear to provide a useful representative in vitro model system to study the physiological and pathologic aspects of the human corneal epithelium. PMID- 9856767 TI - Mitotic inhibition of corneal endothelium in neonatal rats. AB - PURPOSE: Corneal endothelium in humans does not divide to any significant extent after birth; therefore, with age there is a gradual loss of cells. When cell density is reduced to a critical level, the endothelium cannot function to maintain corneal clarity, and the cornea becomes permanently cloudy. Currently, the blindness that results can be treated only by corneal transplantation. The long-term goal is to find methods to stimulate corneal endothelial proliferation in a clinically relevant manner. The first step toward achieving this goal is to identify mechanisms responsible for the induction and maintenance of mitotic inhibition of the corneal endothelium in vivo. During corneal development, the endothelium is formed by migration and proliferation of mesenchymal cells from the ocular periphery. Soon after the monolayer is formed, proliferation ceases. In tissue culture, many cell types cease proliferating upon formation of stable cell-cell and cell-substrate attachments. The goal of the present studies was to determine whether establishment of stable contacts correlates with cessation of endothelial proliferation during corneal development in vivo. METHODS: Corneas from neonatal (days 1, 3, 7, 10, 13, 14, 17, 21, 28, and 42) and adult rats were used for immunolocalization of the following: bromodeoxyuridine (BrdU), an S phase marker; p27kip1 and p21cip1, G1-phase inhibitors; connexin-43 and ZO-1, proteins associated with gap and tight junctions, respectively; Na+/K+-ATPase and beta3-integrin, markers of plasma membrane polarity; and fibronectin and collagen type IV, constituents of Descemet's membrane. Nuclei staining positively for BrdU were counted to determine the relative number of S-phase cells at various times after birth. Marker protein expression and localization were determined by conventional fluorescence microscopy and by confocal microscopy. RESULTS: The number of endothelial cells staining positively for BrdU gradually decreased between postnatal days 1 and 13. After postnatal day 13, positive BrdU staining was no longer detectable. During the first postnatal week, cells stained positively for the G1-phase inhibitor p27kip1 but not for p21cip1. Connexin-43 achieved its mature location by postnatal day 1. ZO-1, Na+/K+-ATPase, beta3 integrin, fibronectin, and collagen type IV achieved their mature localization patterns between postnatal days 14 and 21. CONCLUSIONS: In neonatal rat, corneal endothelial cells are still entering the cell cycle at birth, but cell cycle entry gradually decreases, so that by postnatal day 13 cells are no longer entering the S-phase. The G1-phase inhibitor p27kip1, but not p21cip1, may help mediate this inhibition. Stable cell-cell and cell-substrate contacts gradually form, and monolayer maturation is complete between postnatal days 14 and 21. The results lead to the hypothesis that, in developing rat cornea in vivo, the establishment of stable cell-cell and cell-substrate contacts initiates a cascade of events, mediated by p27kip1, which induces mitotic inhibition in the endothelial monolayer. PMID- 9856768 TI - Characterization of a soluble KGF receptor cDNA from human corneal and breast epithelial cells. AB - PURPOSE: Keratinocyte growth factor (KGF) is a member of the fibroblast growth factor (FGF) family FGF-7. It exhibits potent mitogenic activity for epithelial cells, including corneal and mammary epithelial cells. A messenger RNA has been reported that is generated by alternative splicing of bek that putatively codes only for the extracellular ligand-binding domain of KGF receptor (soluble KGF receptor). In the present study, the expression of the mRNA coding for this alternative bek transcript was examined and the corresponding protein characterized. METHODS: Alternative messenger RNA transcripts were detected in various cell lines or tissues using reverse transcription-polymerase chain reaction (RT-PCR) and RNase protection assay. NIH/3T3 fibroblast cells and 293 kidney embryonic epithelial cells were stably transfected with soluble KGF receptor cDNA and transmembrane KGF receptor cDNA. Soluble KGF receptor protein was produced using a baculovirus-insect expression system. Soluble KGF receptor protein was detected using western and dot blot analyses. Binding assays and cross-linking labeling were used to determine the affinity and specificity of soluble KGF receptor. A mitogenic assay was performed to examine the function of the soluble KGF receptor. RESULTS: The soluble KGF receptor mRNA was primarily expressed in epithelial cells, including cells from the cornea and breast. Cross linking labeling and affinity-binding assays with 125I-KGF showed that the soluble KGF receptor bound KGF (FGF-7) but not FGF-1 or FGF-2. Soluble KGF receptor was detected in the culture medium of cells stably transfected with soluble KGF receptor cDNA but not with transmembrane KGF receptor cDNA, suggesting that the soluble receptor was generated by mRNA splicing and probably not by proteolysis or posttranslational processing. Soluble KGF receptor inhibited KGF binding to transmembrane KGF receptor and DNA synthesis in BALB/MK epidermal keratinocytes in response to KGF, suggesting that soluble KGF receptor expression could provide a mechanism for the cell to downregulate responses to KGF. CONCLUSIONS: A truncated soluble KGF receptor expressed in corneal and other epithelial cells probably functions to downregulate the response of the cell to KGF. PMID- 9856769 TI - Characterization of two corneal epithelium-derived antigens associated with vasculitis. AB - PURPOSE: In a previous investigation into corneal autoimmunity, it was demonstrated that a putative autoantigen, a protein of 66 kDa, present in bovine corneal epithelium, binds circulating autoantibodies in approximately 60% of patients with Wegener's granulomatosis (WG). The aim of the present study was to characterize and identify the 66-kDa protein. METHODS: A purification protocol was established for the 66-kDa protein using standard chromatography techniques. During the purification procedure it became clear that the 66-kDa protein detected in patients' sera was in fact two proteins, both running at 66 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, that eluted in different fractions on DE-52 chromatography columns. These two proteins have been labeled bovine corneal epithelial antigen-A and -B (BCEA-A and BCEA-B). Further investigations of antibody binding have demonstrated that patients' sera bind to either one or the other of these proteins with no cross-reactivity between them. Separated BCEA-A and BCEA-B protein extracts were immunoblotted with 27 WG patients' sera, 10 Churg-Strauss syndrome (CSS) patients' sera, 31 rheumatoid arthritis (RA) patients' sera, and 40 healthy control subjects' sera from the blood bank. RESULTS: Forty-six percent of WG patients' sera had antibodies to one of the 66-kDa antigens, whereas none of the healthy control subjects' sera had 66 kDa antibodies (P < 10(-5)). In the WG group, 31% were positive to BCEA-A (versus controls, P = 0.0023), and 15% were positive to BCEA-B. WG patients with peripheral ulcerative keratitis (PUK) had a significant association with anti BCEA-A antibodies when compared with healthy control subjects (50%, P < 10(-6)). However, in the RA group with no eye disease there was an association with BCEA-A (25%, P = 0.011) but not in the RA group with PUK. The frequency of anti-BCEA-B antibodies was significantly increased in patients with CSS (60%, P < 10(-7)). CONCLUSIONS: In summary, it has been shown that vasculitis patients have antibodies to two 66-kDa corneal antigens and that autoantibodies to these antigens are mutually exclusive. It has also been shown that antibodies to BCEA-B are associated with CSS, whereas BCEA-A antibodies are associated with WG and RA. PMID- 9856770 TI - Alteration of mucin in human conjunctival epithelia in dry eye. AB - PURPOSE: The objective of this study was to determine whether alteration in mucins could be detected in patients with dry eye symptoms by using the monoclonal antibody H185, which recognizes carbohydrate epitopes on mucin molecules. METHODS: Immunohistochemistry and immunoelectron microscopy was used to examine binding of H185 antibody to conjunctival cells obtained by nitrocellulose filter paper stripping (impression cytology). Two study populations were examined. Study I included 22 patients with dry eye symptoms and 13 normal volunteers. Study II included 16 aqueous-deficient dry eye patients and 14 age-matched control subjects. RESULTS: Results of the studies demonstrated significant differences in binding patterns of H185 to conjunctival cells in normal eyes compared with those of patients with dry eye symptoms. In normal eyes, the antibody bound to apical cells in a mosaic pattern, with cells exhibiting either light, medium, or intense binding. A predominant pattern in patients with dry eye symptoms was loss of the mosaic pattern with replacement by a "starry sky" pattern in which there was a lack of apical cell binding (hence, dark sky) but increased binding to goblet cells (hence, stars in the sky). The starry sky pattern correlated with rose bengal staining. CONCLUSIONS: From these studies it is concluded that there is an alteration either in mucin distribution or mucin glycosylation on the surfaces of apical conjunctival cells in dry eye and that glycosylation of goblet cell mucins changes with the disease. PMID- 9856771 TI - Area and depth of surfactant-induced corneal injury predicts extent of subsequent ocular responses. AB - PURPOSE: To correlate area and depth of initial corneal injury induced by surfactants of differing type and irritant properties with corneal responses and outcome in the same animals over time by using in vivo confocal microscopy (CM). METHODS: Six groups of six adult rabbits were treated with anionic, cationic, and nonionic surfactants that caused different levels of ocular irritation. Test materials included slight irritants: 5% sodium lauryl sulfate (SLS), polyoxyethylene glycol monoalkyl ether (POE), and 5% 3-isotridecyloxypropyl bis(polyoxyethylene) ammonium chloride (ITDOP); mild irritants: 5% 3 decyloxypropyl-bis(polyoxyethylene) amine (DOP) and sodium linear alkylbenzene sulfonate (LAS); and a moderate irritant: a proprietary detergent (DTRGT). Ten microliters surfactant were directly applied to the cornea of one eye of each rabbit. Ten untreated rabbits served as control subjects. Area and depth of initial injury was determined by using in vivo CM to measure epithelial thickness, epithelial cell size, corneal thickness, and depth of stromal injury in four corneal regions at 3 hours and at day 1. Area and depth of corneal responses to injury were evaluated at various times from days 3 through 35 by macroscopic grading and quantitative confocal microscopy through-focusing (CMTF). RESULTS: In vivo CM revealed corneal injury with slight irritants to be restricted to the epithelium, whereas the mild and moderate irritants caused complete epithelial cell loss with increasing anterior stromal damage: DOP < LAS < DTRGT. With the slight ocular irritants there was little or no change in corneal thickness or the CMTF intensity profiles. Three hours after treatment, mild and moderate ocular irritants caused a significant increase in corneal thickness, which peaked at day 1 with DOP (483.3+/-80.1 microm) and LAS (572.3+/ 60.0 microm) and day 3 with DTRGT (601.4+/-68.7 microm); returning to normal (similar to control values) by day 7 with DOP and day 35 with LAS and DTRGT. The CMTF intensity profiles also showed significant elevation over that in the anterior stroma, which peaked at day 1 with DOP (14,608+/-4,306 U [U is defined as micrometers X pixel intensity]) and day 3 with LAS and DTRGT (18,471+/-6,581 U and 22,424+/-3,704 U, respectively) and returned toward normal by day 7 with DOP and day 14 with LAS and DTRGT. Elevated CMTF profiles principally reflected the presence of hyperreflective, punctate keratocytes and inflammatory cells at days 1 and 3 and the presence of activated keratocytes at day 7. There was a significant correlation between the elevated CMTF intensity profile and the corresponding macroscopic total score in each eye (r = 0.839; P < 0.001). More important, there was a significant correlation between area and depth of initial stromal injury measured at day 1, regardless of ocular irritant and the stromal response measured by the area under the CMTF intensity profile curve in each cornea (r = 0.87; P < 0.0005). A significant correlation between the area and depth of injury and the area under the corneal thickness curve was also observed in each cornea (r = 0.75; P < 0.0005). CONCLUSIONS: In individual animals, the extent of initial stromal injury correlated with the magnitude of the corneal responses, measured by the change in corneal thickness and the CMTF depth intensity profile. These findings further support the hypothesis that area and depth of injury are the principal factors determining the early responses and eventual repair processes after accidental eye irritation. They also support the proposed use of area and depth of acute injury as a mechanistic correlate to ocular irritation in the development and validation of potential in vitro ocular irritation tests. PMID- 9856772 TI - Bone morphogenic proteins 2 and 4 and their receptors in the adult human cornea. AB - PURPOSE: To examine the expression of transforming growth factor family members bone morphogenic proteins 2 and 4 (BMP2, BMP4), their receptor mRNAs, and BMP2 and BMP4 proteins in the cells of the human cornea. The effects of BMP2 and BMP4 on corneal fibroblast proliferation and apoptosis were also examined. METHODS: Reverse transcription-polymerase chain reaction, immunoprecipitation, and western blot analysis were used to examine mRNA and protein expression in cultured human corneal cells. Immunocytochemistry was applied to examine protein localization in fresh frozen human cornea cells. Stimulation and inhibition of nuclear factor kappaB (NF-kappaB) activation was evaluated by gel shift assay. Apoptosis was examined using trypan blue exclusion, laddering of DNA, CPP32 assay, and transmission electron microscopy. Proliferation was monitored by counting cells. RESULTS: BMP2 and BMP4 mRNAs and proteins were expressed in cultured human corneal epithelial cells, keratocytes, and corneal endothelial cells. BMP2 and BMP4 were detected in each major corneal cell type in fresh frozen human cornea. BMP receptor IA, IB, and II mRNAs were also detected in cultured human corneal epithelial cells, keratocytes, and endothelial cells. BMP2 and BMP4 stimulated activation of NF-kappaB. Actinomycin D and SN50 peptide, but not SN50M control peptide, inhibited NF-kappaB activation in response to BMP2 or BMP4. BMP2 and BMP4 stimulated apoptosis of corneal fibroblast cells when NF-kappaB activation was inhibited with the nonselective inhibitor actinomycin D or selective inhibitor SN50. The nonsteroidal anti-inflammatory agents ketorolac tromethamine and diclofenac sodium augmented the effect of BMP2 on corneal fibroblast apoptosis. BMP2 and BMP4 both stimulated proliferation of corneal fibroblast cells in the absence of inhibitors of NF-kappaB activation. CONCLUSIONS: BMP2, BMP4, and their receptors are expressed in the cells of the adult human cornea. The functions regulated by these cytokines may include keratocyte proliferation and apoptosis. PMID- 9856773 TI - Adult human retinal neurons in culture: Physiology of horizontal cells. AB - PURPOSE: Adult postmortem human retinal neurons in long-term monolayer cultures were recorded to characterize the voltage- and transmitter-gated currents in putative human horizontal cells (HCs). METHODS: Enzymatically and mechanically dissociated human retinal cells were seeded on polylysine and laminin- coated coverslips. Cells were identified by immunocytochemistry with cell type-specific antibodies and recorded with the patch-clamp technique. RESULTS: Immunostaining and responses to voltage steps confirmed the survival of various retinal cell types. Horizontal cells were identified by their specific glutamate-modulated anomalous rectifier K+ current conductance. This identification was further confirmed by subsequent immunolabeling of dye-labeled recorded cells with an anti parvalbumin antibody that selectively stained HCs in frozen human retinal sections. Horizontal cells generated voltage-gated currents classically observed in HCs from fish to mammals: a transient outward K+ current, a sustained outward K+ current, and an L-type (Ca2+ current. Na+ currents were observed in only a few HCs. As in other species, glutamate, gamma-aminobutyric acid (GABA), and glycine generated responses mediated by the activation of kainate/(RS)-alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), GABA(A), and glycine receptors, respectively. CONCLUSIONS: Various human retinal cell populations survive in vitro as indicated by immunolabeling with specific cell markers and by the diversity of responses to voltage steps. Human HCs exhibited extensive physiological similarities to HCs from other vertebrate species and a maintained expression of parvalbumin. These results constitute a comprehensive analysis of voltage- and transmitter-gated currents in a primate retinal neuron and validate the use of long-term monolayer culture of adult human neurons as a novel in vitro model for the study of human vision. PMID- 9856774 TI - Effect of matrix metalloproteinases activity on outflow in perfused human organ culture. AB - PURPOSE: To test the hypothesis that extracellular matrix turnover, mediated by the matrix metalloproteinases, modulates aqueous humor outflow facility in a human outflow model. METHODS: Matrix metalloproteinase activity was manipulated and outflow facility evaluated using perfused human anterior segment organ culture. Purified matrix metalloproteinases, tissue inhibitors of metalloproteinases (TIMPs), and several families of synthetic inhibitors of matrix metalloproteinases were added to the perfusion medium. Matrix metalloproteinase expression was increased by adding recombinant interleukin (IL) 1alpha. Kinetic inhibition analysis was conducted for stromelysin, gelatinase A, and gelatinase B with the various inhibitors. Live-dead staining was used to evaluate culture viability. RESULTS: Increasing metalloproteinase activity, by adding purified metalloproteinases or by inducing their expression by IL-1alpha treatment, increased outflow facility. Inhibition of endogenous trabecular metalloproteinase activity using TIMP or several families of synthetic metalloproteinase inhibitors reduced outflow rates. The elevation and the reduction of outflow rates were reversible, with changes requiring 1 to 3 days. Kinetic enzyme inhibition analysis produced 50% inhibitory concentration values for these inhibitors that were compatible with the concentration ranges for outflow inhibition. CONCLUSIONS. The ability of several specific matrix metalloproteinase inhibitors to reduce outflow facility implies that endogenous extracellular matrix turnover by these enzymes was required for the maintenance of trabecular outflow resistance, at least in this human culture model. These observations provide support for the hypothesis that controlled extracellular matrix turnover is important in the regulation of aqueous humor outflow facility. PMID- 9856775 TI - Analysis of immunoregulatory cytokines in ocular fluid samples from patients with uveitis. AB - PURPOSE: To investigate the T-helper cell cytokine profiles in two well-defined clinical uveitis entities caused by an infectious mechanism. METHODS: Cytokines (interleukin [IL]-2, IL-4, IL-6, IL-10, and interferon [IFN]-gamma) were measured in ocular fluid samples obtained from patients with herpes simplex- or varicella zoster virus-induced acute retinal necrosis (ARN; n = 17) and toxoplasma chorioretinitis (n = 27) using enzyme-linked immunosorbent assay techniques. The data were compared with data for 51 control samples taken during cataract surgery (n = 10), vitrectomy in diabetic retinopathy (n = 10), eye bank eyes (n = 10) and with samples from patients with "autoimmune" uveitis (n = 21). RESULTS: Interleukin-6 was detected in 44 of 51 control samples and 43 of 44 eyes of patients with uveitis. The highest levels in the control samples were detected in 9 of 10 vitreous samples from patients with diabetic retinopathy (mean, 648 pg/ml). In 8 of 10 samples taken from patients during cataract surgery and in 7 of 10 eye bank eyes the amount of IL-6 was significantly lower (mean, 10 pg/ml and 136 pg/ml, respectively). Interleukin-6 levels in patients with ARN (mean, 1436 pg/ml) were significantly higher than in those with toxoplasma chorioretinitis (mean, 272 pg/ml). Interleukin-2 was detected in one of the samples from patients with toxoplasma chorioretinitis (1105 pg/ml) and in three samples from the control subjects suffering from Fuchs' heterochromic anterior uveitis (mean, 752 pg/ml). No IL-4 (<2 pg/ml) was detected either in patient or control samples. Interferon-gamma could be detected in 7 of 17 ARN patients (range, 277-3483 pg/ml), in 13 of 27 samples from patients with toxoplasma chorioretinitis (range, 12-250 pg/ml), and in 1 of 21 of the samples from control subjects with uveitis (31 pg/ml) but was absent in nonuveitic control samples. Interleukin-10 was detected in 10 of 17 ARN patients (range, 29-3927 pg/ml), in 13 of 27 samples from patients with toxoplasma chorioretinitis (range, 4-67 pg/ml), and in only 3 of 51 control samples (6 pg/ml, 16 pg/ml, and 20 pg/ml). CONCLUSIONS: Various immunoregulatory cytokines (IL-6, IL-10, and IFN-gamma) were detected in ocular fluid samples from patients with uveitis. A separate role for either a T-helper type 1 or T-helper type 2 response in the pathogenesis of clinical uveitis could not be proven. PMID- 9856776 TI - Role of mucosal IgA in the resistance to Acanthamoeba keratitis. AB - PURPOSE: To determine whether oral immunization with Acanthamoeba castellanii antigens elicits mucosal antibodies of the IgA isotype and whether mucosal antibodies affect parasite adhesion to the corneal epithelium. METHODS: Chinese hamsters were immunized with 100 microg aqueous Acanthamoeba antigen mixed with cholera toxin (Ac-CT) and subsequently challenged with parasite-laden contact lenses that were applied to abraded corneal surfaces. Tears and stool samples were examined for the presence of Acanthamoeba-specific IgA antibodies by enzyme linked immunosorbent assay (ELISA). The effect of mucosal antibody on trophozoite binding to corneal epithelium and viability of trophozoites was examined in vitro. RESULTS: Hamsters immunized orally with Ac-CT showed significantly lower infection rates than did control groups (21.4% versus 72.6%). ELISA analysis of mucosal specimens showed the presence of parasite-specific IgA in stool samples and tears from hamsters orally immunized with Ac-CT, but not in control animals. In vitro assays showed that anti-Acanthamoeba IgA did not affect parasite viability. However, mucosal anti-Acanthamoeba IgA profoundly inhibited (>75%) the binding of parasites to corneal epithelial cells in vitro. CONCLUSIONS: Oral immunization with Ac-CT induces the production of parasite-specific IgA in mucosal secretions and prevents corneal infection. Mucosal antibody does not affect the viability of Acanthamoeba trophozoites but seems to prevent infection by inhibiting parasite binding to the corneal epithelium. PMID- 9856777 TI - SPARC deficiency leads to early-onset cataractogenesis. AB - PURPOSE: To determine the role of SPARC (secreted protein, acidic, and rich in cysteine) in cataractogenesis by examining mice deficient in a matricellular protein SPARC. METHODS: Mice were rendered SPARC-deficient by a targeted disruption of the gene. Slit-lamp microscopy and histology were used to examine the eyes of SPARC-null and wild-type mice from birth to 14 months of age. RESULTS: SPARC-null mice developed opacities in the posterior cortex of the eye as early as 1.5 months after birth. The diffuse cataracts appeared to progress toward the anterior cortex and reached maturity in many animals by 3.5 months of age. Early stages of cataractogenesis in SPARC-null mice included inhibition of normal lens fiber cell differentiation, degeneration of fiber cells, vacuole formation at the equator, and liquefaction of the cortex. No cataracts were detected in wild-type mice up to the age of 8 months. CONCLUSIONS: The early onset of cataracts in SPARC-null mice establishes that the gene is essential to the maintenance of lens transparency. PMID- 9856778 TI - Apoptosis in the rat lens after in vivo threshold dose ultraviolet irradiation. AB - PURPOSE: To investigate DNA damage in the rat lens after in vivo close-to threshold exposure to ultraviolet radiation (UVR). METHODS: Sprague-Dawley rats received 5 kJ/m2 UVR (lambdaMAX = 300 nm, lambda0.5 = 10 nm) unilaterally for 15 minutes. Animals were killed at 1, 6, and 24 hours and at 1 week after exposure. DNA-strand breaks were investigated in sagittal paraffin sections using the TdT dUTP terminal nick-end labeling (TUNEL) technique and propidium iodide for counterstaining. Other lenses were prepared for transmission electron microscopy (TEM). RESULTS: TUNEL-positive nuclei were found at only 24 hours after UVR exposure. About one tenth of the epithelial cell nuclei were TUNEL positive, and affected cells were scattered over the entire epithelium. No TUNEL-positive cells were found at 1 or 6 hours or at 1 week after UVR exposure or in the nonexposed lenses. TEM verified the occurrence of programmed cell death and showed the breakdown of the apoptotic cells by adjacent cells. No signs of necrosis were found. CONCLUSIONS: Threshold-dose UVR induces programmed cell death that peaks 24 hours after exposure and involves the entire epithelium. Dead cells are removed from the epithelium by phagocytosis. PMID- 9856779 TI - 3-FG as substrate for investigating flux through the polyol pathway in dog lens by 19F-NMR spectroscopy. AB - PURPOSE: To investigate flux through the polyol pathway in the dog lens by 19F nuclear magnetic resonance (19F-NMR) spectroscopy, using 3-fluoro-3-deoxy-D glucose (3-FG) as a substrate. METHODS: 3-FG metabolism was monitored by 19F-NMR analysis. Dog lenses were incubated in Dulbecco's modified Eagle's medium containing 10 mM 3-FG. Enzymatic reductase and dehydrogenase activities were spectrophotometrically determined, whereas the analyses of 3-FG metabolites were conducted by 19F-NMR analysis. Aldose reductase (AR) was immunohistochemically localized in dog lens with antibodies raised against dog kidney AR. RESULTS: 19F NMR spectra indicate that incubation of purified dog lenses AR with 3-FG results in the formation of 3-fluoro-3-deoxy-D-sorbitol (3-FS) and that incubation of dog liver sorbitol dehydrogenase (SDH) with 3-FS results in the formation of 3-fluoro 3-deoxy-D-fructose (3-FF). This confirms that 3-FG is metabolized to 3-FF by the polyol pathway enzymes. The affinity (Km) of AR for 3-FG is approximately 20-fold better than that for D-glucose, whereas the Km of SDH for 3-FS was fourfold less than for D-sorbitol. 3-FG in cultured dog lenses is metabolized primarily to 3 FS; however, small amounts of 3-FF and 3-fluoro-3-deoxy-D-gluconic acid (3-FGA) are also formed. 3-FS formation was reduced by the AR inhibitor AL 1576, and 3-FF formation was eliminated by the SDH inhibitor CP-166,572. In dog lens epithelial cells cultured with 3-FG, only 3-FS is formed. Similarly, only 3-FS is formed when lens capsule containing primarily epithelial lens contaminated with superficial epithelial cells was incubated in 3-FG. Similar incubation of the remaining cortex resulted primarily in the formation of 3-FS and 3-FGA. This enzymatic distribution was confirmed by spectrophotometric activity analysis and the immunohistochemical localization of AR. CONCLUSIONS: The data confirm that flux through the polyol pathway primarily results in sorbitol accumulation. The absence of fructose and gluconic acid from cultured lens epithelium suggests that the epithelial cells primarily contain AR, whereas differentiated fiber cells also contain SDH and glucose dehydrogenase. PMID- 9856780 TI - Human age-related cataract and lens epithelial cell death. AB - PURPOSE: To evaluate the importance of lens epithelial cell death in age-related cataract. To determine whether the large percentage of terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive lens epithelial cells previously reported in human capsulotomy specimens results from apoptosis or necrosis. METHODS: Capsulotomy specimens from patients who had undergone cataract surgery and epithelia from cataractous lenses of eye bank eyes were compared with epithelia from noncataractous lenses of eye bank eyes. DNA fragmentation was assayed using the TUNEL method. Cell membrane integrity was tested using a fluorescent stain for DNA, BOBO-3, that is excluded from living cells. Cell proliferation was assayed by labeling with 5-bromo-2'-deoxyuridine (BrdU). The number of cells in different regions of the lens epithelium was measured by digital imaging and computerized counting of nuclei after staining with methyl green. RESULTS: TUNEL-positive cells were sometimes detected adjacent to denuded regions of capsulotomy specimens, especially when epithelia were not fixed immediately after surgery. TUNEL-stained cells usually stained with BOBO-3, indicating loss of plasma membrane integrity. No BrdU-labeled cells were detected in capsulotomy specimens. Cell density in cataractous lens epithelia was similar to that in normal lens epithelia. In cataractous lenses from eye bank eyes, cell density in the region of the epithelium overlying the cataract was higher than cell density in the region of the epithelium overlying the transparent part of the lens. No correlation was found between cell density and cataract severity or between cell density and age. CONCLUSIONS: TUNEL staining of lens epithelial cells in capsulotomy specimens most likely results from necrotic cell death caused by damage during or soon after cataract surgery. Loss of cells from the lens epithelium, by apoptosis or other mechanisms of cell death, does not seem to play a major role in age-related cataract formation. PMID- 9856781 TI - Nocturnal elevation of intraocular pressure in young adults. AB - PURPOSE: To distinguish 24-hour (circadian) and postural effects on intraocular pressure (IOP) in healthy young adults. METHODS: Thirty-three volunteers were housed in a sleep laboratory for 1 day under a strictly controlled 16-hour light and 8-hour dark environment. Sleep was encouraged in the dark period. Intraocular pressure was measured in each eye every 2 hours using a pneumatonometer. Researchers used night-vision goggles to perform IOP measurements in the dark, while the subject's light exposure was minimized. In the first group of 12 subjects, measurements were taken with subjects in the sitting position during the light-wake period and supine during the dark period. In the second group of 21 subjects, all IOP measurements were taken with the subjects supine. RESULTS: Average IOP was significantly higher in the dark period than in the light-wake period in both groups. The lowest IOP occurred in the last light-wake measurement, and the peak IOP occurred in the last dark measurement. The trough peak difference in IOP was 8.2+/-1.4 mm Hg (mean +/- SEM) in the first group. Intraocular pressure changed sharply at the transitions between light and dark. In the second group, the trough-peak IOP difference was 3.8+/-0.9 mm Hg. Intraocular pressure changed gradually throughout the 24-hour period. In comparison with the sitting IOP in the first group, the supine IOP in the second group was significantly higher during the light-wake period. CONCLUSIONS: Circadian rhythms of IOP were shown in young adults, with the peaks occurring in the late dark period. A nocturnal IOP elevation can appear independent of body position change, but change of posture from upright to recumbent may contribute to the relative nocturnal IOP elevation. PMID- 9856782 TI - Reevaluation of Cl-/HCO3- exchange in cultured bovine corneal endothelial cells. AB - PURPOSE: To determine the apical versus basolateral polarity of the putative anion exchanger in cultured bovine corneal endothelial cells (BCECs) and to examine the influence of Cl--dependent membrane potential (Em) changes on HCO3- transport. METHODS: BCECs grown on permeable supports were used for independent perfusion of apical and basolateral surfaces. Intracellular pH (pHi) was measured using the fluorescent dye BCECF. Relative changes in Em were measured using the fluorescent dye bis-oxonol. Western blot analysis was used to detect immunoreactivity against the anion exchanger (AE1 or AE2). RESULTS: Cl- removal from apical and basolateral surfaces produced cellular alkalinization (apical side, 0.07 pH units; basolateral side, 0.06 pH units; both sides, 0.20 pH units). Application of 100 microM H2-4,4'-diisothiocyanatodihydrostilbene-2,2'-disulfonic acid (DIDS), an anion exchange inhibitor, on the apical side produced an alkalinization (0.02 pH units) followed by acidification (-0.05 pH units), whereas basolateral H2DIDS caused a substantial acidification (-0.16 pH units). In the absence of Na+, Cl- removal from the apical side caused a transient alkalinization (0.03 pH units) followed by a return to baseline; Cl- removal from the basolateral side caused a small (-0.03) acidification. In Na+-free Ringer, apical H2DIDS produced a transient alkalinization (0.02 pH units), whereas basolateral exposure had no effect. 5-Nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), N-phenylanthranilic acid (DPC), and niflumic acid (50-200 microM), known Cl- channel blockers, produced cellular acidification in control Ringer. Niflumic acid hyperpolarized Em and inhibited depolarization after Cl- removal. Western blot analysis failed to detect AE2 expression in cultured BCECs. However, fresh BCECs produced a trace response. CONCLUSIONS: Physiological activity of an apical anion exchanger is weak in cultured BCECs. Cultured BCECs have significant Cl- conductance. Thus, cellular alkalinization after Cl- removal is caused primarily by depolarization of Em, which drives HCO3- influx through the basolateral electrogenic Na+:nHCO3- cotransporter. In contrast with cultured BCECs, AE2 may be present in fresh cells. PMID- 9856783 TI - Multiple genetic loci modify risk for retinoblastoma in transgenic mice. AB - PURPOSE: Forty percent of cases of retinoblastoma, a childhood malignancy of the retina, are linked to the inheritance of a mutant allele of the retinoblastoma susceptibility gene Rb1. Tumor penetrance varies among carriers in different family pedigrees, indicating that other genetic factors may modify risk for occurrence of retinoblastoma. This study was undertaken to determine whether multiple genetic loci modify the risk for retinoblastoma in mice. METHODS: A line of alphaAcry-HPV16E6/E7 transgenic mice expressing the human papillomavirus type 16 E6 and E7 oncogenes (HPV-16 E6 and E7) ectopically in the retina was characterized. E6 and E7 proteins bind to and inactivate the cellular tumor suppressor proteins p53 and Rb, respectively. RESULTS: Retinoblastomas developed rarely when the alphaAcry-HPV16E6/E7 transgene was maintained on the FVB background, but tumors arose with high frequency on C57BL/6 X FVB and C3H x FVB F1 hybrid backgrounds. The incidence of retinoblastoma in the LHbeta-TAG transgenic mice, which express simian virus 40 large tumor antigen (SV40 T-ag), was also influenced by the FVB and C57BL/6 backgrounds. Resistance of the alphaAcry-HPV16E6/E7 FVB mice to retinoblastoma mapped in part to the retinal degeneration (rd) locus. However, multiple genetic experiments indicate that resistance to retinoblastoma depends on additional loci in FVB mice. CONCLUSIONS: Multiple cellular genes can modify risk for retinoblastoma in mice. PMID- 9856784 TI - A new method for noninvasive optical sectioning of the chorioretinal vasculature. AB - PURPOSE: To report a new method for optical sectioning of the chorioretinal vasculature to improve the visualization of vascular abnormalities due to chorioretinal eye diseases. METHODS: An imaging system was developed for optical sectioning of the vasculature called chorioretinal optical sectioning (CROS). CROS consists of projecting a laser beam at an angle on the retina after injection of a fluorescent dye and viewing the fluorescence. On the fluorescence optical section (FOS) image, the vasculature of the retina and choroid appear laterally displaced according to their depth location. The laser beam is scanned over a 2 X 2-mm area to generate 40 FOS images, each spatially separated by 50 microm on the retina. Optical section images of the vascular layers are constructed from the series of FOS images. RESULTS: CROS permitted optical separation of vascular layers in living eyes. Optical section images of normal and laser-photocoagulated retinas had higher contrast than conventional angiography because of the separation of the fluorescence from the overlapping layers and allowed enhanced visualization of vascular abnormalities. CONCLUSIONS: CROS enhances the visualization of the retinal and subretinal vasculature and promises to be a beneficial tool for evaluation of chorioretinal diseases. PMID- 9856785 TI - Use of the ARPE-19 cell line as a model of RPE polarity: basolateral secretion of FGF5. AB - PURPOSE: To determine the polarity of fibroblast growth factor 5 (FGF5) secretions from retinal pigment epithelium (RPE) cells and to examine the viability and utility of the ARPE-19 cell line as a model for the study of RPE polarity. METHODS: Influenza infection and adenovirus-mediated gene transfer were used to deliver and express genes encoding influenza hemagglutinin (HA), p75-NTR (a neurotrophin receptor), low-density lipoprotein (LDL) receptor (LDLR), and FGF5 in confluent monolayers of ARPE-19 cells. The localization of HA, p75-NTR, and LDLR was determined by confocal microscopy. Domain selective biotinylation assays were used to quantitatively determine the polarities of p75-NTR and LDLR. The secretion of FGF5 into the apical and basal media of ARPE-19 cultures was examined by immunoblot analysis of conditioned media. RESULTS: Hemagglutinin and p75-NTR were found to be localized on the apical surface of infected and transduced ARPE-19 cells. In contrast, LDLR was associated preferentially with the basolateral membrane of ARPE-19 cells. Biotinylation studies indicated that 84% of p75-NTR was present on the apical surface, and 79% of LDLR was basolaterally polarized. Over the course of 6 hours, more than 90% of the total secreted FGF5 protein accumulated in the basolateral media. CONCLUSIONS: ARPE-19 cells exhibit a polarized distribution of cell surface markers when examined by either confocal microscopy or surface-labeling assays. This indicates that the ARPE-19 cell line is a valid model for studies of RPE cell polarity. FGF5, a secreted protein normally produced by RPE cells, is accumulated preferentially in the basal media after only 6 hours, suggesting that it is vectorially secreted from the basolateral surface of ARPE-19 cells. PMID- 9856786 TI - Docosahexaenoic acid promotes differentiation of developing photoreceptors in culture. AB - PURPOSE: The purpose of this work was to study the effects of diverse fatty acids on the composition, metabolism, differentiation, and characteristics of opsin expression in retina photoreceptors. METHODS: Cultures of rat retinal neurons were incubated with or without 22:6 n-3, 22:5 n-3, 20:4 n-6, 18:1 n-9, and 16:0, labeled and unlabeled. RESULTS: In photoreceptor cells incubated with 22:6 n-3 and 22:5 n-3, the proportions of these fatty acids in phospholipids increased four- to sixfold. The remaining fatty acids did not change lipid acyl chain composition. The labeled fatty acids were all actively esterified in neuronal lipids, particularly in phosphatidylcholine. Addition of unlabeled 22:6 n-3 did not affect the distribution among lipids of the other fatty acids but displaced [3H]20:4 n-6 from phosphatidylcholine and phosphatidylethanolamine. These results suggest that retinal neurons have specific mechanisms for processing fatty acids of different lengths and degrees of unsaturation and that 22:6 n-3 incorporation takes priority. Of all fatty acids, 22:6 n-3 was the most effective in promoting photoreceptor differentiation. In 22:6-sufficient photoreceptors, new apical processes formed, the expression of opsin augmented, and its localization improved, concentrating in the apical processes of the cells. CONCLUSIONS: The advancement in differentiation selectively elicited by 22:6 correlates with the fact that 22:6 n-3, but none of the other fatty acids, delays significantly the onset of apoptosis in photoreceptors in culture. The synthesis of 22:6-containing phospholipid molecules could be required for the proper localization of opsin. This could contribute to furthering the differentiation of photoreceptors, preventing their apoptosis, and extending their survival. PMID- 9856787 TI - Stimulus duration, neural adaptation, and sweep visual evoked potential acuity estimates. AB - PURPOSE: Results in several studies have suggested that the visual evoked potential (VEP) amplitude can vary with stimulus duration. The purpose of this study was to determine whether acuity estimates obtained by extrapolation of the sweep VEP are altered by this adaptation effect. METHODS: Sweep VEP data were obtained from 16 healthy observers under binocular viewing conditions. Data were acquired with a commercially available VEP unit using standard electrode recording techniques. Three sweeps (high spatial frequencies, medium spatial frequencies, and low spatial frequencies) were run. The subjects' visual acuity at the monitor distance was 6/6 for the high spatial frequency sweep. For the medium and low spatial frequency sweeps, the subjects were dioptrically blurred to 6/15 (medium spatial frequencies) or 6/30 (low spatial frequencies) at the monitor distance. Each sweep consisted of six spatial frequencies (contrast 80%; temporal frequency (TF) = 7.5 Hz; screen luminance = 100 candela [cd]/m2). For each spatial frequency, the stimulus duration was 8 seconds, partitioned into 1 second bins. A minimum of eight sweeps were obtained per subject. An acuity estimate was obtained for each second's data by fitting a line to the high spatial frequencies (excluding noise) and extrapolating this line to the x-axis. With this technique, estimates could not be obtained for 29 of 384 possible acuities. RESULTS: The sweep VEP acuities for the 16 subjects did not change significantly over the 8 seconds of data collection for the high, medium, or low spatial frequency sweep (repeated measures analysis of variance [ANOVA]: high, P = 0.25; medium, P = 0.50; low, P = 0.23). In any given subject, there was a 1- to 2-octave range in acuity estimates over the 8 seconds of stimulus presentation (high, 1.23+/-0.417 octaves; medium, 1.41+/-0.593 octaves; low, 1.52+/-0.475 octaves; mean +/- SD). CONCLUSIONS: These results suggest that there is not a significant change in sweep VEP acuity estimates over an 8-second stimulus presentation. Thus, neural adaptation does not significantly affect the clinical use of the sweep VEP. PMID- 9856789 TI - Ascorbate in the corneal epithelium of diurnal and nocturnal species. AB - PURPOSE: To compare the amount of ascorbic acid in the corneal epithelium of various species to unveil possible differences between diurnal and nocturnal mammals. METHODS: Ascorbic acid was determined by high-performance liquid chromatography, using an LC-10 system (Shimadzu, Kyoto, Japan). RESULTS: Diurnal animals show a higher ascorbate concentration in the corneal epithelium than nocturnal animals. Ascorbate concentration is higher in the corneal epithelium than in the matching aqueous humor in diurnal and nocturnal species. The highest ascorbate concentration is found in the corneal epithelium of the reindeer. CONCLUSIONS: Ascorbate level in the corneal epithelium seems to vary in accordance with ambient radiation exposure of the respective species, just as in the aqueous humor. Both phenomena are regarded as environmental adaptations, and the ascorbic acid is suggested as protecting against photokeratitis and as acting as an ultraviolet filter for internal eye structures. PMID- 9856788 TI - Involvement of interleukin-1beta-converting enzyme in apoptosis of irradiated retinoblastomas. AB - PURPOSE: To investigate whether interleukin-1beta-converting enzyme (ICE), a mammalian homologue of the Caenorhabditis elegans cell death gene ced-3, is involved in gamma-irradiation-induced apoptosis (programmed cell death) of human retinoblastoma cells. METHODS: The induction of apoptotic cell death in human retinoblastoma cell lines WERI-Rb-1 and Y79 by gamma-irradiation was determined with a modified 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide colorimetric assay and the DNA-binding fluorochrome bis (benzimide) trihydro chloride (Hoechst 33258) staining. The change of ICE protein level in tumor cells during apoptosis was determined by immunoblotting assay. Whether the specific tetrapeptide ICE inhibitor Ac-YVAD-CMK affected gamma-irradiation-induced apoptosis in tumor cells was also examined. The effect of ICE overexpression on tumor cells was evaluated by a transient transfection assay using ICE expression vector. RESULTS: Gamma-irradiation inhibited the cell viability of WERI-Rb-1 and Y79 cells in a dose-dependent manner and induced apoptosis. The protein level of ICE was remarkably enhanced after the treatment. The apoptotic cell death induced by gamma-irradiation was suppressed by the tetrapeptide ICE inhibitor Ac-YVAD CMK. Moreover, overexpression of ICE induced apoptosis in tumor cells. CONCLUSIONS: These findings suggest that ICE may play an important role in gamma irradiation-induced apoptosis in retinoblastoma cells. Transfer of the ICE gene induces apoptosis in these cells without gamma-irradiation. PMID- 9856790 TI - pH dependence of corneal oxygen consumption. AB - PURPOSE: To determine whether corneal acidosis, which occurs during contact lens wear, alters corneal O2 consumption (QO2) and if so, whether increased ion transport activity could contribute to altered QO2 during acidosis. METHODS: PO2 was measured, using the phosphorescence quenching of Pd-meso-tetra-(4 carboxyphenyl) porphine, in an airtight chamber that held a trephined rabbit cornea. The rate of change in chamber PO2 was used as a measure of QO2. QO2 was measured at pH 7.5 and then at either pH 6.7, 7.1, or 7.3. Measurements of QO2 at pHs 7.5 and 6.7 were repeated in the presence of 0.5 mM amiloride and 0.5 mM ouabain. RESULTS: When pH was changed from 7.5 to 6.7, 7.1, or 7.3, O2 consumption increased by a factor of 1.80+/-0.11 (+/-SE), 1.65+/-0.12, and 1.44+/ 0.06, respectively. Amiloride (0.5 mM) and ouabain (0.5 mM) inhibited 50% and 65%, respectively, of the increase in QO2 at pH 6.7. CONCLUSIONS: Corneal acidosis leads to increased QO2 in a dose-dependent manner. The increased QO2 is in part secondary to the activation of pH regulatory mechanisms, including Na+/H+ exchange, which then stimulates Na+/ K+-ATPase activity. These findings indicate that contact lens-induced acidosis can exacerbate corneal hypoxia and related complications. PMID- 9856791 TI - Biometric confirmation of the Hirschberg ratio in strabismic children. AB - PURPOSE: In the Hirschberg eye position test, the ratio of strabismic angle to decentration of the corneal reflex is dependent on two biometric parameters of the eye: the radius of the corneal curvature and the depth of the anterior chamber. This study was designed to confirm whether the Hirschberg conversion ratio (HR) previously determined for adults can be used for children of various ages despite structural growth of the eye. METHODS: For 262 eyes of 131 children with strabismus (age range, 6 months to 11 years), the radius of the corneal curvature was measured with an auto-keratometer and the anterior chamber depth with an A-scan ultrasound unit under general anesthesia before the surgery. Using these measurements, the HR was computed on the basis of a geometric model. RESULTS: The calculated HR was constant across the age range, and the mean+/-SD was 19.9+/-1.9 prism diopters/mm (95% confidence interval, 16.1-23.6 prism diopters/mm). The ratios for the two eyes in each subject showed good correlation (R = 0.854, P = 0.0001). Neither of the biometric measurements was significantly correlated with age, although considerable scatter of the measurements was observed. CONCLUSIONS: These results indicate that the averaged HR can be applied in children regardless of the patient's age, although intersubject variance of the ratio should be taken into account. PMID- 9856792 TI - Cytokine expression in a rat model of Staphylococcus aureus endophthalmitis. AB - PURPOSE: To examine the ability of viable Staphylococcus aureus to induce the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, cytokine induced neutrophil chemoattractant (CINC), and interferon (IFN)-gamma after intravitreal injection. METHODS: Experimental rat eyes were injected with a 25 microl volume of approximately 80 colony-forming units of viable S. aureus; control eyes received sterile saline. Eyes were graded daily for signs of clinical inflammation and were removed 6, 24, 48, and 72 hours after injection. One group was prepared for histologic analysis, and vitreous was removed from the other group for cytokine analysis, using standard enzyme-linked immunosorbent assay procedures. RESULTS: TNF-alpha, IL-1beta, CINC, and IFN-gamma were detected in experimental vitreous samples at increased levels that peaked at 24 hours. TNF alpha, IL-1beta, and CINC declined at 48 hours, but IFN-gamma remained elevated. At 72 hours, levels returned to baseline. Statistically significant elevations of TNF-alpha, IL-1beta, and CINC were detected in experimental samples at 24, but not at 6 and 48 hours compared with levels in saline control samples (P < 0.03). A statistically significant increase in IFN-gamma was detected at 24 and 48 hours compared with control levels (P < 0.03). In experimental animals, clinical inflammation and inflammatory cells peaked at 24 hours, persisted at 48 hours, and began to decline thereafter. Neutrophils were the predominant inflammatory cell detected at 24 (72.3% of cells) and 48 (60.1%) hours. By 72 hours, the total number of inflammatory cells had decreased by 75.0%, and the cellular infiltrate had changed so that neutrophils equaled monocytes-macrophages. CONCLUSIONS: S. aureus induced the expression of TNF-alpha, IL-1beta, CINC, and IFN-gamma. The time course of these cytokine levels could account for the clinical inflammatory responses and the entry and decline of vitreous cells in this model of bacterial endophthalmitis. PMID- 9856793 TI - Expression profiles of P2-receptor isoforms P2Y1 and P2Y2 in the rat lens. AB - PURPOSE: To verify at the molecular level that P2 receptors are expressed in the lens and to determine their expression profiles. METHODS: The reverse transcription-polymerase chain reaction (RT-PCR) was used to screen rat lens epithelial and fiber cells for the expression of the ionotropic P2X2 receptor and the G-protein-linked receptor isoforms P2Y1 and P2Y2. Northern blot analysis was used to confirm the level of expression of P2Y1 and P2Y2. The profile of P2 receptor isoform expression in the lens was identified using an indirect RT-PCR in situ hybridization procedure on paraffin sections of whole rat eyes. RESULTS: P2X2-receptor transcripts could not be detected in lens epithelial or fiber cells. P2Y1- and P2Y2-receptor transcripts were detected in lens fiber cells but not in the epithelial cells. Their expression profiles were maximal in the lens cortex and mostly overlapped each other, except that transcripts for the P2Y2 receptor isoform appeared earlier in the not yet fully elongated fiber cells in the lens bow region. CONCLUSIONS: The molecular data support physiological evidence previously reported by others that P2Y receptors are expressed in the lens and identifies cortical fiber cells as the principal site of expression. PMID- 9856794 TI - Twenty-four-hour change in axial length in the rabbit eye. AB - PURPOSE: To study the 24-hour changes in axial length, lens thickness, and anterior chamber depth in rabbits and to examine the role of ocular sympathetic activity on these changes. METHODS: Young adult rabbits were entrained to a daily 12-hour light-12-hour dark cycle. Axial length, lens thickness, and anterior chamber depth were measured using ultrasonic techniques. In the first group of 12 rabbits, measurements were taken in the middle light phase and in the early dark phase. In the second group of 12 rabbits, measurements were taken in constant dark every 2 hours for a period of 24 hours. The latter group of rabbits underwent unilateral transection of the cervical sympathetic trunk. Three to four weeks later, axial length, lens thickness, and anterior chamber depth were measured again in constant dark every 2 hours for 24 hours. RESULTS: Under the light- dark condition, axial length and anterior chamber depth were larger in the early dark phase than in the middle light phase. Lens thickness changed in the opposite direction. Under the constant-dark condition, axial length and anterior chamber depth changed gradually during the 24-hour period. The trough appeared in the late subjective light phase, and the peak appeared in the late subjective dark phase. Lens thickness remained relatively constant. In the eyes with decentralized ocular sympathetic nerves, 24-hour changes in axial length and anterior chamber depth occurred. However, magnitudes of nocturnal enlargement were relatively smaller than those in the intact eyes. Although larger in the decentralized eyes, lens thickness appeared unchanged in either eye for 24 hours. CONCLUSIONS: Consistent 24-hour changes in axial length and anterior chamber depth occur in young adult rabbits. These changes are driven endogenously. Significant portions of the nocturnal enlargements of axial length and anterior chamber depth are unrelated to ocular sympathetic activity. PMID- 9856795 TI - Transgenic mice with pigmented intraocular tumors: tissue of origin and treatment. AB - PURPOSE: To describe the cell of origin, tumor progression, light and electron microscopic appearance, immunohistochemical properties, and response to frequently used anticancer therapies in two transgenic models of intraocular melanoma. METHODS: Two lines of transgenic mice that develop pigmented intraocular tumors were produced with the SV40 T and t antigens under the control of the mouse tyrosinase gene. Tumors were sequentially studied and characterized by light microscopy, electron microscopy, and immunohistochemistry stains. Tumor response to two cycles of dacarbazine was assessed on the basis of tumor size in one group of animals. Response to external beam irradiation was measured by survival time in other animals. RESULTS: Two lines of transgenic mice developed bilateral intraocular tumors with complete penetrance and without primary cutaneous melanomas. Tumors developed first in the retinal pigment epithelial layer, with subsequent retinal and choroidal invasion, extraocular extension, and metastasis. Tumors stained positive for S-100, HMB-45, and Fas-ligand. Electron microscopy revealed polarization of tumor cells with basement membrane formation, microvilli, immature melanosomes, and abundant endoplasmic reticulum. Dacarbazine significantly reduced tumor size in these mice, and a trend toward dose-dependent decrease in survival was found with external beam irradiation. CONCLUSIONS: Tumors developed from the retinal pigment epithelium. Their histology and growth, however, closely resembled that of human choroidal melanoma. This model may be a useful tool for future studies of endogenous primary pigmented tumors limited to the eye. Response to standard therapies suggests it can serve as a model with which to evaluate therapeutic modalities. PMID- 9856797 TI - Photomechanical transcutaneous delivery of macromolecules. AB - Transcutaneous drug delivery has been the subject of intensive research. In certain situations, rapid transcutaneous delivery is very desirable. A mechanical (stress) pulse generated by a single laser pulse was shown to transiently increase the permeability of the stratum corneum in vivo. The barrier function of the stratum corneum recovers within minutes. The increased permeability during these few minutes allows macromolecules to diffuse through the stratum corneum into the viable epidermis and dermis. Macromolecules (40 kDa dextran and 20 nm latex particles) were deposited into the skin using a photomechanical pulse generated by a single 23 ns laser pulse. This treatment can potentially be utilized in therapies that currently require occlusive dressings for hours or day(s). PMID- 9856796 TI - Comparison of genetic profiles between primary melanomas and their metastases reveals genetic alterations and clonal evolution during progression. AB - To examine for the genetic basis of metastatic progression in cutaneous melanoma, we have compared loss of heterozygosity (LOH) of several selected chromosome regions that are implicated in the initiation and progression of melanoma, and alterations of the p16INK4a gene in 14 pairs of primary tumor and synchronous or asynchronous metastasis excised from the same patients. The most frequent genetic alteration during metastatic progression detected was the loss of p16INK4a protein expression (four of 14 cases), whereas no somatic p16INK4a gene mutations were found in any primary or metastatic tumors. LOH analyses showed that most of the chromosome losses including 6q, 8p, 9p, 9q, and 18q were shared between primary tumors and their metastases. Nevertheless, LOH of 6q and 11q and LOH of 7q not detected in primary tumors were, respectively, observed in two lymph node metastases. These results suggest that loss of p16INK4a protein expression (but not p16INK4a gene mutation) and the losses of chromosome arms 6q, 7q, and 11q play an important role in the acquisition of metastatic potential in sporadic melanoma. Furthermore, comparison of genetic profiles between the primary tumor and its metastasis revealed in several cases that heterogenous tumor cell populations might already exist at the early stage of tumorigenesis and evolve independently in the primary tumor and its metastasis, strongly suggesting that metastatic progression of sporadic melanoma is not accounted for by a linear progression model. PMID- 9856798 TI - Effect of tenascin-C deficiency on chemically induced dermatitis in the mouse. AB - Tenascin-C is a large extracellular matrix glycoprotein characterized by its spatiotemporal expression during embryogenesis, carcinogenesis, and wound healing. Many in vitro studies on tenascin-C have revealed its multifunctional properties; however, disruption of the tenascin-C gene did not reveal any obvious abnormalities during development, and its function in vivo remains unclear. Here, we investigated whether tenascin-C is involved in inflammatory dermatitis in adults by studying chemically induced dermatitis in tenascin-C knockout mice. An epicutaneous application of a hapten, 2,4-dinitrofluorobenzene, to the ear skin of BALB/CA mice resulted in inflammation and induced the expression of tenascin C. In congenic tenascin-C knockout mice, the dermatitis occurred more severely than in wild-type mice; infiltration of polymorphonuclear cells in knockout mice persisted longer than in wild-type mice, and the elastosis-like disorganized extracellular matrix was also seen in the ear. These results suggest that tenascin-C plays a role in vivo in inflammatory responses in the skin, and that the genetic background has profound effects on the function of tenascin-C in mouse dermatitis. PMID- 9856799 TI - Human melanocytes and keratinocytes exposed to UVB or UVA in vivo show comparable levels of thymine dimers. AB - Epidemiology shows a relationship between solar exposure and all types of skin cancer. Understanding the mechanisms of skin cancer requires knowledge of the photomolecular events that occur within the relevant epidermal cell types in vivo. Studies to date have focused on UVR-induced DNA lesions in keratinocytes, the majority epidermal cell population which gives rise to most skin cancers. Malignant melanoma, arising from melanocytes (5%-10% of epidermal cells), accounts for most skin cancer deaths. We report on new techniques to detect DNA photolesions in human epidermal melanocytes in situ. Previously nonexposed buttock skin of volunteers of skin types I/II was exposed to clinically relevant doses of narrow bandwidth UVB (300 nm) and UVA (320 nm, 340 nm, 360 nm) radiation. Biopsies were taken immediately afterwards and processed for routine histology. Microscope sections were prepared and double-stained with fluorescent tagged monoclonal antibodies for thymine dimers and melanocytes. UVR dose response curves for dimer levels within melanocyte nuclei were determined by image analysis and compared with dimer levels in adjacent basal cell keratinocytes. Our data show that UVB and UVA readily induce thymine dimers in melanocytes at levels that are comparable with those found in adjacent keratinocytes. This new technique will enable melanocyte specific studies, such as DNA repair kinetics, to be done in vivo. PMID- 9856800 TI - Do hair bulb melanocytes undergo apoptosis during hair follicle regression (catagen)? AB - The fate of the hair follicle pigmentary unit during the cyclical involution of anagen hair follicles is unknown. Using the C57BL/6 mouse model for hair research, hair follicle melanocytes were examined during the anagen-catagen transformation, comparing spontaneous and pharmacologically induced catagen development. This study shows that both spontaneous catagen and dexamethasone induced catagen display similar changes in the pigmentary unit. Catagen hair follicles exhibited pigment incontinence in the dermal papilla and in selected outer root sheath keratinocytes. Melanocytes deleted by apoptosis were detected in spontaneous catagen and, more commonly, in dexamethasone-induced catagen, and were identified using transmission electron microscopy by the presence of free premelanosomes in affected cells lacking epithelial specializations, and by the colocalization of TUNEL positivity and tyrosinase-related protein-1 immunoreactivity. By contrast, cyclophosphamide-induced catagen was characterized by the initial retention of melanogenic and dendritic melanocytes in the presence of widespread keratinocyte apoptosis. Melanocyte incontinence and the ectopic distribution of melanin were more severe than in the other forms of catagen. Whereas much of this melanin was extruded, via the hair canal, to the skin surface, hair follicle-derived pigment was also detected within the epidermis, probably derived from pigment-carrying migrating outer root sheath keratinocytes from the proximal hair follicle. Thus, apoptosis may account, at least in part, for the loss of melanogenic melanocytes during spontaneous catagen. Although dexamethasone-induced catagen may provide a useful model for general hair pigmentation research, catagen induced by cyclophosphamide offers an interesting model for studying the response, and relative resistance, of melanocytes to chemical injury. PMID- 9856801 TI - Analysis of apoptotic cell death in human hair follicles in vivo and in vitro. AB - We analyzed changes of growth and apoptotic cell death in human hair follicles. In anagen hair follicles, terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate-biotin nick labeling-positive cells were observed in the keratogenous zone of the upper bulb matrix, the inner root sheath, and the companion layer of the outer root sheath. DNA ladder formation was also detected in anagen hair follicles. In catagen hair follicles, the lower bulb matrix cells around the dermal papilla and the outer layer cells of the outer root sheath became strongly positive, showing that apoptosis in catagen hair is distinct from that in anagen hair. We also confirmed the mRNA expression of four caspases (caspase-1, caspase-3, caspase-4, and caspase-7) in anagen hair follicles by reverse transcriptase-polymerase chain reaction and in situ hybridization. When human anagen hair follicles were cultured in the presence of transforming growth factor-beta or tumor necrosis factor-alpha in the serum-free medium, transforming growth factor-beta but not tumor necrosis factor-alpha induced catagen-like morphologic changes, which were indistinguishable from normal catagen hair follicles. Tumor necrosis factor-alpha, however, strongly inhibited the elongation of the hair shaft in a dose-dependent manner, accompanied by abnormal morphology and increased cell death in the bulb matrix cells. Our results suggest that apoptosis in hair follicles involves two different types. One is related to the terminal differentiation of follicular epithelial cells in anagen hair. The other occurs as a major driving force to eliminate the distinct portion of epithelial components in catagen hair. Furthermore, this study strongly indicates that the transforming growth factor-beta pathway is involved in the induction of catagen phase in human hair cycle. PMID- 9856802 TI - A novel human type II cytokeratin, K6hf, specifically expressed in the companion layer of the hair follicle. AB - In an attempt to identify new members of the human type II hair keratin family by means of 3'- and 5'-RACE methods and cDNA from anagen hair follicles, we detected a sequence that encoded a hitherto unknown type II cytokeratin. The novel cytokeratin comprises 251 amino acids and exhibits the highest sequence homology with K5. Comparative one- and two-dimensional western blots of keratins from anagen hair bulbs, containing or not containing the outer and inner root sheaths (ORS/IRS), and from footsole epidermis with an antibody against the new cytokeratin, revealed its comigration with K6 and its expression in the ORS/IRS complex. We have therefore named the new cytokeratin K6hf, to distinguish it from the various K6 isoforms and to indicate its expression in the hair follicle. Both in situ hybridization with a K6hf-specific cRNA probe and indirect immunofluorescence with the K6hf antibody showed that K6hf is exclusively expressed in the so-called "companion layer" of the hair follicle, a single layered band of flat and vertically oriented cells between the cuboidal ORS cells and the IRS that stretches from the lowermost bulb region to the isthmus of the follicle. Concomitant K17 and K16 expression studies showed that besides suprabasal ORS cells, these cytokeratins are sequentially expressed subsequent to K6hf in companion cells above the hair bulb. Our study confirms the view of a vertically oriented companion layer differentiation. The clearly delayed K17 and K16 expression relative to that of K6hf in companion cells most probably excludes these keratins as possible type I partners of K6hf and suggests the existence of a still unknown type I partner of its own. Thus, not only morphologically but also biochemically, the companion layer is different from the ORS and can therefore be regarded as an independent histologic compartment of the hair follicle. PMID- 9856803 TI - Localization of rat FGF-5 protein in skin macrophage-like cells and FGF-5S protein in hair follicle: possible involvement of two Fgf-5 gene products in hair growth cycle regulation. AB - It has been reported that the gene for murine fibroblast growth factor-5 (Fgf-5) is expressed in the rat hair follicle and that this expression may be associated with catagen induction (Hebert et al, 1994). In this study, we analyzed the Fgf-5 gene product in skin because the gene generates two mRNA that translate into the FGF-5 protein and a short form of the FGF-5 protein (FGF-5S) as a result of an alternative splicing (Hattori et al, 1996; Ozawa et al, 1996). Indeed, we detected both types of FGF-5 mRNA in rat skin samples. Two monoclonal anti-FGF-5 antibodies, one (E723) being specific for FGF-5 long-form protein and the other (B2B6) being reactive with both FGF-5 and FGF-5S proteins, were used to locate these proteins by immunohistochemistry. Staining of the rat skin revealed that only the B2B6 antibody reacted with hair follicles and that both antibodies reacted with macrophage-like round cells, suggesting that the product of the Fgf 5 gene in the hair follicle is FGF-5S. The immunoreactivity of the FGF-5S protein increased during early anagen VI and decreased rapidly during catagen. The density of FGF-5-positive macrophage-like cells in the dermis increased during anagen and decreased during catagen and telogen, whereas the density of these cells in the panniculus adiposus did not change during anagen and increased during catagen and telogen. There was no apparent association between the density of FGF-5-positive macrophage-like cells and that of FGF-5-negative, dendritic macrophage-like cells. Thus, the results suggest the possible involvement of FGF 5S in the hair follicle in anagen VI and catagen development and that the density of FGF-5-positive macrophage-like cells may also be associated with the hair growth cycle. PMID- 9856804 TI - Alterations in cholesterol sulfate and its biosynthetic enzyme during multistage carcinogenesis in mouse skin. AB - Recent evidence suggests that cholesterol sulfate may be an important second messenger involved in signaling epidermal differentiation in skin. The activity of cholesterol sulfotransferase (Ch-ST) is increased during squamous differentiation of keratinocytes and is believed to be a marker enzyme for terminal differentiation. The primary objective of this study was to examine changes in levels of cholesterol sulfate (CS) and activity of its biosynthetic enzyme, Ch-ST, during multistage carcinogenesis in mouse skin. Using SENCAR mice, we determined the activity of Ch-ST in normal epidermis, in tumor promoter treated epidermis, in epidermis during wound healing, and in mouse skin tumors generated by initiation-promotion regimens. A single topical application of tumor promoters led to significantly elevated levels of Ch-ST activity and of CS. Epidermal Ch-ST activity was also elevated during wound healing. Dramatic increases in CS levels and in the activity of Ch-ST were found in nearly all of the papillomas and squamous cell carcinomas examined. The increased levels of CS and activity of Ch-ST in tumor promoter-treated epidermis were accompanied by increased transglutaminase-I activity. In contrast, transglutaminase I activity was not elevated in primary papillomas or squamous cell carcinomas. Finally, Ch ST activity was significantly elevated in the epidermis of newborn HK1.ras transgenic mice, whereas transglutaminase I activity did not correlate with Ch-ST activity in these mice. These results demonstrate that diverse tumor-promoting stimuli all produce elevated CS levels and Ch-ST activity and that CS levels and Ch-ST activity were constitutively elevated in both papillomas and squamous cell carcinomas. The data also suggest a mechanism for upregulation of Ch-ST in skin tumors involving activation/upregulation of Ha-ras. PMID- 9856805 TI - The similarity of action spectra for thymine dimers in human epidermis and erythema suggests that DNA is the chromophore for erythema. AB - The location of DNA photodamage within the epidermis is crucial as basal layer cells are the most likely to have carcinogenic potential. We have determined the action spectra for DNA photodamage in different human epidermal layers in situ. Previously unexposed buttock skin was irradiated with 0.5, 1, 2, and 3 minimal erythema doses of monochromatic UVR at 280, 290, 300, 310, 320, 340, and 360 nm. Punch biopsies were taken immediately after exposure and paraffin sections were prepared for immunoperoxidase staining with a monoclonal antibody against thymine dimers that were quantitated by image analysis. Dimers were measured at two basal layer regions, the mid and the upper living epidermis. The slopes of dose response curves were used to generate four action spectra, all of which had maxima at 300 nm. Dimer action spectra between 300 and 360 nm were independent of epidermal layer, indicating comparable epidermal transmission at these wavelengths. Furthermore, we observed 300 nm-induced dimers in dermal nuclei; however, there was a marked effect of epidermal layer between 280 and 300 nm, showing relatively poor transmission of 280 and 290 nm to the basal layer. These data indicate that solar UVB (approximately 295-320 nm) is more damaging to basal cells than predicted from transmission data obtained from human epidermis ex vivo. The epidermal dimer action spectra were compared with erythema action spectra determined from the same volunteers and ultraviolet radiation sources. Overall, these spectral comparisons suggest that DNA is a major chromophore for erythema in the 280-340 nm region. PMID- 9856806 TI - Living skin substitutes: survival and function of fibroblasts seeded in a dermal substitute in experimental wounds. AB - The healing of full-thickness skin defects requires extensive synthesis and remodeling of dermal and epidermal components. Fibroblasts play an important role in this process and are being incorporated in the latest generation of artificial dermal substitutes. We studied the fate of fibroblasts seeded in our artificial elastin/collagen dermal substitute and the influence of the seeded fibroblasts on cell migration and dermal substitute degradation after transplantation to experimental full-thickness wounds in pigs. Wounds were treated with either dermal substitutes seeded with autologous fibroblasts or acellular substitutes. Seeded fibroblasts, labeled with a PKH-26 fluorescent cell marker, were detected in the wounds with fluorescence microscopy and quantitated with flow cytofluorometric analysis of single-cell suspensions of wound tissue. The cellular infiltrate was characterized for the presence of mesenchymal cells (vimentin), monocytes/macrophages, and vascular cells. Dermal substitute degradation was quantitated by image analysis of wound sections stained with Herovici's staining. In the wounds treated with the seeded dermal substitute, fluorescent PKH-26-labeled cells were detectable up to 6 d and were positive for vimentin but not for the macrophage antibody. After 5 d, flow cytofluorometry showed the presence of 3.1 (+/-0.9) x 10(6) (mean +/- SD, n = 7) PKH-26-positive cells in these wounds, whereas initially only 1 x 10(6) fluorescent fibroblasts had been seeded. In total, the percentage of mesenchymal cells minus the macrophages was similar after 5 d between wounds treated with the seeded and the acellular substitutes. In the wounds treated with the seeded substitute, however, 19.5% of the mesenchymal cells were of seeded origin. Furthermore, the rate of substitute degradation in the seeded wounds was significantly lower at 2-4 wk after wounding than in wounds treated with the acellular substitute. Vascular in growth and the number of infiltrated macrophages were not different. In conclusion, cultured dermal fibroblasts seeded in an artificial dermal substitute and transplanted onto full-thickness wounds in pigs survived and proliferated. The observed effects of seeded fibroblasts on dermal regeneration appeared to be mediated by reducing subcutaneous fibroblastic cell migration and/or proliferation into the wounds without impairing migration of monocytes/macrophages and endothelial cells. Moreover, the degradation of the implanted dermal substitute was retarded, indicating a protective activity of the seeded fibroblasts. PMID- 9856807 TI - Induction of SLPI (ALP/HUSI-I) in epidermal keratinocytes. AB - Secretory leukocyte protease inhibitor (SLPI) is a small, cationic protein that is known to be constitutively expressed by several glandular epithelia. SLPI inhibits leukocyte-derived proteinases, has anti-HIV-1, antibacterial, and anti fungal properties, and interferes with the induction of synthesis of proinflammatory mediators in monocytes and macrophages. We now report that at both the mRNA and the protein level, SLPI shows inducible expression in a nonglandular epithelium. A weak expression of SLPI was found in the stratum granulosum of adult normal human epidermis; however, in lesional psoriatic epidermis and in migrating keratinocytes of healing wounds, a strong cytoplasmic staining was seen in the suprabasal keratinocytes. Remarkably, in the dermis adjacent to SLPI-expressing keratinocytes, SLPI was found extracellularly associated with elastin fibers, whereas the dermis in normal skin was negative. In cell culture, SLPI was hardly expressed in monolayers of proliferating keratinocytes. Differentiating cultures with a phenotype of normal skin expressed low levels of SLPI, whereas cultures with a regenerative/psoriatic phenotype expressed high levels. Functional studies with recombinant SLPI indicated that its antibacterial spectrum and potency are distinct from other anti-microbial peptides such as lysozyme and defensins. In view of the multiple functions of SLPI and the inducibility, we propose that it acts as an important first line defence mechanism in cutaneous injury. PMID- 9856808 TI - MIG is a dominant lymphocyte-attractant chemokine in lichen planus lesions. AB - Dense accumulation of mononuclear cells (lymphocytes >> macrophages) in the dermal-epidermal interface and a T cell-mediated cytotoxic reaction against basal keratinocytes are hallmarks of lichen planus lesions. In this study, we focused on the chemotactic signals responsible for the selective recruitment of these cells. Using in situ hybridization and immunohistochemistry, the expression and localization of the lymphocyte-and/or monocyte/macrophage-attractant CC chemokines macrophage chemoattractant protein-1 (MCP-1), regulated on activation, normal T cell expressed, and secreted (RANTES), macrophage inflammatory protein 1alpha and -1alpha (MIP-1alpha/beta), I-309 and the CXC chemokines monokine induced by interferon-gamma (MIG), interferon-gamma-inducible protein-10 (IP-10), interleukin-8 (IL-8), epithelial-derived neutrophil attractant-78, and growth related oncogene-alpha were investigated. Strong mRNA expression of MIG, IP-10, and MCP-1 and moderate mRNA expression of RANTES and MIP-1alpha were detected exclusively within foci characterized by strong infiltration with CD3+ lymphocytes (CD4+ cells > CD8+ cells) and CD68+ macrophages. All other chemokines investigated were minimally expressed or absent. With more than 11% of total cells strongly expressing MIG transcripts, this selectively lymphotactic chemokine was by far the dominant chemokine and thus may significantly contribute to the inflammatory reaction in lichen planus lesions. According to the mRNA expression profiles, MIG, IP-10, and MCP-1 were expressed by both basal keratinocytes above and mononuclear cells within the inflammatory foci. Our findings indicate that a set of chemokines composed of IP-10, MCP-1, RANTES, MIP 1alpha, and especially MIG contributes to the cytokine network and preferential trafficking of mononuclear cells to the interface region of lichen planus lesions. PMID- 9856809 TI - A serotonin-like immunoreactivity is present in human cutaneous melanocytes. AB - Immunohistochemistry was applied in the investigation of the possible existence of serotonin in human skin. It was found that epidermal melanocytes express a serotonin-like immunoreactivity. The immunoreactivity was associated with both the cytoplasm and the cellular membrane, though the latter was only found in certain cells. The serotonin anti-serum labeled the same cells as NKI-beteb, which is known as a reliable marker of melanocytes. Blocking experiments showed that both serotonin and NKI-beteb have different epitopes in the melanocytes. In in vitro studies, serotonin-like immunoreactivity appeared in approximately 90% of cultured human melanocytes, and was found both in the cytoplasm and also in the nuclei. Thus, we believe the melanocytes to be the origin of serotonin (or a serotonin-like molecule) in the skin. PMID- 9856810 TI - Interaction of BP180 (type XVII collagen) and alpha6 integrin is necessary for stabilization of hemidesmosome structure. AB - The hemidesmosome is a multimolecular complex that integrates the extracellular matrix with the keratin cytoskeleton and that stabilizes epithelial attachment to connective tissue. A 180 kDa protein (BP180, type XVII collagen), first identified by its reactivity with autoantibodies in the serum of patients with a blistering skin disease called bullous pemphigoid (BP), is a transmembrane component of the hemidesmosome with a collagen-like extracellular domain. Here, using recombinantly expressed molecules and the yeast two-hybrid assay, we have identified alpha6 integrin as a BP180-binding partner. The association between specific domains of the BP180 and alpha6 integrin molecules is inhibited by a 14 mer peptide, whose sequence is identical to amino acid residues 506-519 in the noncollagenous region of the ectodomain of the BP180 molecule, as well as by antibodies raised against this peptide. The 14 mer peptide sequence is part of an epitope recognized by autoantibodies that are pathogenic in BP. In vivo, when 804G cells are plated into medium containing the same peptide, they fail to assemble hemidesmosomes. Furthermore, although BP180 and certain cytoplasmic components of the hemidesmosome colocalize in the peptide-treated cells, they are aberrantly distributed and fail to show extensive association with (alpha6beta4 integrin. Taken together, our results indicate that BP180 is a novel transmembrane ligand of the alpha6beta4 integrin heterodimer. In addition, our data provide support for the possibility that BP180 and alpha6 integrin interaction is not only mediated by the BP epitope but is necessary for hemidesmosome formation. PMID- 9856811 TI - Increased expression of heat-shock protein 47 is associated with overproduction of type I procollagen in systemic sclerosis skin fibroblasts. AB - Heat-shock protein 47 (HSP47) is a collagen-binding stress protein that is thought to act as a collagen-specific molecular chaperon during the biosynthesis and secretion of procollagen. In this study we examined the expression of HSP47 mRNA and protein in systemic sclerosis (SSc) skin fibroblasts. HSP47 mRNA and protein levels were significantly higher in fibroblast cultures from SSc patient involved skin samples than in fibroblasts from normal skin from healthy individuals, as assessed by northern blot and immunoblot analyses, respectively. SSc cultured fibroblasts with increased levels of HSP47 mRNA and protein showed high expression of type I procollagen. By in situ hybridization, SSc skin had a higher number of fibroblasts with high HSP47 and procollagen alpha1(I) mRNA levels than normal skin, and the distribution of HSP47 mRNA was similar to that of procollagen alpha1(I) mRNA. We also investigated the effects of cytokines on the expression of HSP47 in normal cultured fibroblasts. Transforming growth factor-beta1 and interleukin-4 increased HSP47 mRNA and protein levels, whereas interferon-gamma reduced HSP47 expression. The same pattern of cytokine-regulated expression was observed for type I procollagen levels. These results indicate that HSP47 expression is closely associated with that of type I procollagen in skin fibroblasts, and that increased expression of HSP47 may be involved in the abundant production of type I procollagen by SSc fibroblasts. PMID- 9856812 TI - Epidermal expression of collagenase delays wound-healing in transgenic mice. AB - A vital characteristic of skin is its ability for wound repair in response to injury. A transient elevation of matrix metalloproteinases (MMP) in the epidermal and dermal compartments of healing wounds implicates the MMP family of enzymes in the regulation of events important to injury repair. Transgenic mice expressing human interstitial collagenase (MMP-1) in the epidermis were used to perturb the regulation of this proteinase in order to examine the role of epidermal collagenase during wound healing. The relative healing potential of collagenase transgenic mice and wild-type littermates was assessed by measurements of the wound area during closure of full-thickness wounds. Transgenic mice exhibited a 2 3 d delay in the time required to reach 50% closure of 6 mm wounds. Histologic analysis of the transgenic wound bed revealed the retarded migration of the epithelium across the open wound. The results are consistent with the hypothesis that control of collagenase (MMP-1) expression is important for re epithelialization during wound healing and indicate that collagenase regulation is critical to the kinetics of normal wound closure. PMID- 9856813 TI - Anti-tyrosinase-related protein-2 immune response in vitiligo patients and melanoma patients receiving active-specific immunotherapy. AB - Several melanosome glycoproteins have been shown to be antigenic in humans. Correlation of antigen-specific immune responses in patients with the autoimmune disease vitiligo, therapy-induced hypopigmentation, and cutaneous melanoma has not been well studied. We examined antibody responses to a melanocyte autoantigen, tyrosinase-related protein-2 (TRP-2), as it is highly expressed in cutaneous melanoma and melanocytes. TRP-2 recombinant protein was synthesized for western blot and affinity anti-TRP-2 enzyme-linked immunosorbent assay. We demonstrated that patients with malignant melanoma, vitiligo, and active-specific immunotherapy-induced depigmentation had significant anti-TRP-2 IgG titers. The highest level of anti-TRP-2 IgG response was found in vitiligo patients. Induction and enhancement of anti-TRP-2 IgG responses were observed in melanoma patients treated with a polyvalent melanoma cell vaccine containing TRP-2. Active specific immunotherapy could induce and/or augment the TRP-2 IgG antibody titers. Melanoma patients who developed hypopigmentation and had improved survival after polyvalent melanoma cell vaccine had significantly augmented anti-TRP-2 antibody responses compared with patients with poor prognosis. This study demonstrates that TRP-2 autoantigen is immunogenic in humans. TRP-2 antibody responses provide a linkage between autoimmune responses by vitiligo patients and melanoma patients responding to immunotherapy who have induced hypopigmentation. PMID- 9856814 TI - MCP-1 and MIP-1alpha are most efficient in recruiting T cells into the skin in vivo. AB - To analyze T cell recruitment by chemokines in vivo, we used SCID mice grafted with human skin onto their backs and human T cells into their peritoneal cavities. rh MIP-1alpha and rh MCP-1, as well as mouse MCP-1, attracted significant amounts of human T cells into human skin grafts, whereas effects of rh RANTES, rh SDF-1, or rh IP-10 were minimal. Human T cells were found in a striking perivascular position exclusively at sites of human endothelium. None of the chemokines recruited human T cells across mouse endothelial barriers. For control purposes, chemokines were injected into the abdominal wall directly above the peritoneal cavity, where human T cells had been injected and were not subjected to flow. At this site all tested chemokines attracted human T cells to an equal extent. In summary, our experiments point towards an important role for MCP-1 and MIP-1alpha as T cell chemoattractants for skin inflammatory conditions. PMID- 9856815 TI - Telomerase is not an epidermal stem cell marker and is downregulated by calcium. AB - The ribonucleoprotein complex telomerase, which was found to be active in germ line, immortal, and tumor cells, and in cells from continuously renewing normal tissues such as epidermis or bone marrow, is thought to be correlated with an indefinite life span. Therefore, it has been postulated that in the normal tissues, telomerase activity may be restricted to stem cells, the possible precursors of tumor cells. Here, we demonstrate that a 56% enriched population of epidermal stem cells exhibited less telomerase activity than the more actively proliferating transit amplifying cells, which are destined to differentiate after a finite number of cell divisions. Thus telomerase is not a stem cell marker. In human epidermis we found a heterogeneous expression of the telomerase RNA component (hTR) within the basal layer, with clusters of hTR-positive cells showing variable activities. Histone-3 expressing S-phase basal cells were distributed evenly, illustrating that hTR upregulation may not strictly be correlated with proliferation. We further show for human epidermal cells that differentiation-dependent downregulation of telomerase correlates with Ca++ induced cell differentiation and that increasing the amount of Ca++ but not Mg++ or Zn++ reduced telomerase activity in a dose-dependent manner in a cell-free system (differentiation-independent). Furthermore, addition of ethyleneglycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid completely reversed this Ca++-induced inhibition. These data indicate that Ca++ is not only an important regulator of epidermal differentiation but also a key regulator of telomerase. PMID- 9856816 TI - Expression of interleukin-12 is increased in psoriatic skin. AB - Although the precise underlying pathomechanisms of psoriasis have not been fully elucidated, previous reports suggest that T helper 1-type cytokines are critically involved in the pathogenesis of this disease. Interleukin-12 (IL-12), a heterodimeric cytokine, has been suggested to play a major role in the development of T helper 1 cell responses. In this study, the presence of IL-12 mRNA and protein was investigated in normal human skin as well as nonlesional and lesional psoriatic skin. Messenger RNA levels were determined in biopsy specimens by a standard and a nested reverse transcriptase-polymerase chain reaction method. Additionally, IL-12 protein expression was analyzed in situ by immunohistochemistry using an antibody recognizing IL-12 p70. Whereas specific transcripts for IL-12 p35 were reproducibly detected without any significant differences in all samples, enhanced IL-12 p40 mRNA signals were only found in lesional psoriatic skin as compared with normal and nonlesional psoriatic skin. Furthermore, immunoreactivity for IL-12 p70 was markedly increased in the psoriatic skin lesions and was predominantly expressed on mononuclear cells in the dermis. In conclusion, our data suggest a critical role for IL-12 in promoting and maintaining T cell activation and inducing T helper 1-type cytokines such as interferon-gamma in psoriasis. We speculate that IL-12 might be a key cytokine in the pathogenesis of psoriasis. PMID- 9856817 TI - Induction of inducible nitric oxide synthase and its corresponding tetrahydrobiopterin-cofactor-synthesizing enzyme GTP-cyclohydrolase I during cutaneous wound repair. AB - Recent work has suggested a possible role of nitric oxide, a free radical gas, during the wound healing process. In this study we investigated the regulation of inducible nitric oxide synthase (iNOS) and GTP-cyclohydrolase I (GTP-CH I), the rate-limiting enzyme in the biosynthesis of the iNOS cofactor (6R) 5,6,7,8 tetrahydrobiopterin (6-BH4), during the repair process. We found a similar time course of induction of iNOS and GTP-CH I expression, whereas absolute expression levels were different for both genes. Immunohistochemical analysis revealed colocalization of iNOS and GTP-CH I proteins in the wound. Systemic treatment with glucocorticoids significantly altered the expression levels of iNOS and GTP CH I. Expression of iNOS and GTP-CH I was suppressed by glucocorticoids in normal, and to a much greater extent in wounded skin. Furthermore, a role of nitric oxide as a novel mediator of gene regulation during healing is suggested by the demonstration of nitric oxide-mediated induction of vascular endothelial growth factor expression in keratinocytes. These findings may provide an explanation for the beneficial effects of orally supplemented L-arginine on wound healing, and suggest that a disturbed induction of iNOS and GTP-CH I expression may at least partially underlie the wound healing defect seen in glucocorticoid treated animals. PMID- 9856818 TI - Coordinated induction of inducible nitric oxide synthase and GTP-cyclohydrolase I is dependent on inflammatory cytokines and interferon-gamma in HaCaT keratinocytes: implications for the model of cutaneous wound repair. AB - Recently we demonstrated a strong expression of inducible nitric oxide synthase (iNOS) and GTP-cyclohydrolase I (GTP-CH I) in the basal keratinocytes of the epidermis adjacent to the wound and of the hyperproliferative epithelium during wound healing. To identify possible mediators of iNOS and GTP-CH I expression during this process, we analyzed the regulation of iNOS and GTP-CH I expression in cultured human keratinocytes. We found a large and long lasting coinduction of iNOS and GTP-CH I expression upon simultaneous treatment of quiescent cells with inflammatory cytokines interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma, but not with serum growth factors. The stimulatory effect of interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma is strongly synergistic on iNOS and GTP-CH I expression, because these factors alone stimulated GTP-CH I expression, although to a much lesser extent. Furthermore, iNOS mRNA levels are not influenced at all by stimulation with IL-1beta and revealed only a weak induction after treatment with tumor necrosis factor-alpha and interferon-gamma. Induction of iNOS and GTP-CH I gene expression upon cytokine and interferon-gamma exposure is independent of de novo protein synthesis. Because these cytokines are present at the wound site, they might be responsible for iNOS and GTP-CH I induction during cutaneous repair. Serum, which is released upon hemorrhage, is likely to play no stimulatory role in iNOS and GTP-CH I induction during wound healing. PMID- 9856819 TI - Identification and quantitation of interferon-gamma producing T cells in psoriatic lesions: localization to both CD4+ and CD8+ subsets. AB - Interferon-gamma (IFN-gamma) produced by lesional T cell clones is critical for the induction into G1 of the cell cycle by psoriatic keratinocyte stem cells; however, direct data demonstrating psoriatic lesional T cell subset IFN-gamma expression, and quantitation at a single cell level to calculate in vivo proportions, are lacking. In this study, using flow cytometry of freshly isolated normal and psoriatic lesional T cells from keratome biopsies, we found elevated CD3+, CD4+, and CD8+ T cells in all compartments of psoriatic skin, compared with normals. Using Brefeldin A to induce short-term intracellular accumulation of IFN gamma in T cells capable of IFN-gamma production, we found that 90% of psoriatic patients have IFN-gamma-producing T cells at a greater proportion of their CD3+ cells than normals, with a mean of 16%+/-3%, as compared with 4%+/-2% in normal epidermis (p = 0.01). Expressed as density in the tissue, the IFN-gamma+ CD3+ cell number in psoriatic epidermis was 97+/-22 per mm2 surface area, as compared with 4.4+/-1.8 per mm2 of normal epidermis (p = 0.002). Thus, the total number of IFN-gamma+CD3+ T cells in the skin of a patient with 20% involvement is estimated to be 3.9 x 10(8). CD4+ and CD8+ IFN-gamma+ T cells were both elevated in psoriatic epidermis (p = 0.04 and p = 0.008, respectively) relative to normal skin. In the dermis, only 44% of patients demonstrated a higher percentage of IFN gamma-producing T cells than did normals (p = 0.1), possibly indicating dilution, in some patients, by fresh infiltrating T cells. Interleukin-4 was not found by a combination of flow cytometry, reverse transcriptase-polymerase chain reaction, western blot, and immunoprecipitation. In conclusion, a significant portion of lesional T cells in psoriasis are IFN-gamma producing, without interleukin-4. The increased numbers of both IFN-gamma+CD4+ and IFN-gamma+CD8+ T cells indicate that both CD4+ and CD8+ IFN-gamma+ T cells are present in appropriate anatomic locations to sustain the lesional pathology. PMID- 9856820 TI - Increased type 2 cytokine expression by both CD4+ CD45RO+ T cells and CD8+ CD45RO+ T cells in blood circulation is associated with high serum IgE but not with atopic dermatitis. AB - Type 2 cytokines, such as interleukin-4 (IL-4) and IL-13, are associated with immunoglobulin E (IgE) production. This association has also been observed in CD8+ T cells from patients infected with leprosy and human immunodeficiency virus (HIV). Using intracellular cytokine staining and flow cytometry, the cytokine profile [IL-2, IL-4, IL-10, IL-13, and interferon (IFN)-gamma] of both CD4+ and CD8+ memory/effector T cells circulating in atopic dermatitis (AD) patients was investigated at the single cell level. The levels of type 2 cytokines in CD4+ T cells or CD8+ T cells in AD patients with high levels of serum IgE (AD-H), low levels of serum IgE (AD-L), and healthy controls were compared. Increased production of IL-4 and IL-13 in both CD4+ CD45RO+ T cells and CD8+ CD45RO+ T cells after 4 h in vitro stimulation with phorbol 12-myristate 13-acetate and ionomycin, was more prominent in AD-H patients than in AD-L patients or healthy controls, whereas IFN-gamma-producing CD4+ CD45RO+ T cells and CD8+ CD45RO+ T cells were relatively diminished in AD-H patients. CD4+ T cells and CD8 + T cells from AD-H patients, cultured for 48 h with phorbol 12-myristate 13-acetate and ionomycin, released larger amounts of IL-4 and IL-13 but smaller amounts of IFN gamma than both types of cells from AD-L patients or healthy controls. In addition, when stimulated with immobilized anti-CD3 monoclonal antibody (MoAb) and anti-CD28 MoAb, CD4+ CD45RO+ T cells and CD8+ CD45RO+ T cells from AD-H patients contained more IL-4-producing cells but fewer IFN-gamma-producing cells compared with healthy controls. Finally, spontaneous mRNA expression of IL-4 in blood CD8+ CD45RO+ T cells isolated from AD-H patients was increased, as determined by reverse transcriptase-polymerase chain reaction. Therefore, in AD patients with high IgE levels, type 2 cytokine (IL-4 and IL-13) expression is associated with IgE production, in both CD4+ CD45RO+ T cell and CD8+ CD45RO+ T cell subsets. PMID- 9856821 TI - First comparative delineation of the T cell receptor repertoire in primary and multiple subsequent/coexisting metastatic melanoma sites. AB - At present, very little is known about the types and heterogeneity of T cell responses and immunodominant epitopes of melanoma-associated antigens at coexisting sites of primary melanoma and metastatic lesions. To address this issue, we compared the T cell receptor (TCR) gene usage, complementary determining region 3 diversity, and melanoma-associated antigens expression patterns of primary and metastatic melanoma specimens from three patients with partially homologous HLA class-1 types. Results obtained showed an overall predominance of a very limited number of TCRV regions with AV13 and BV14 being most frequently overexpressed. Sequencing of the dominating TCR transcripts confirmed the restricted usage of certain TCR specificities and, in two of the three patients, identified several identical TCR clonotypes at more than one metastatic site. Nevertheless, we failed to detect TCR transcripts that were common to all tumor deposits in a given patient and, within the majority of coexisting metastases, tumor-infiltrating lymphocytes preferentially used individual site-specifically expanded TCR beta-chain VJ segment combinations. This occurrence of individual responses simultaneously executed at and influenced in their specificity by the different sites of tumor growth, has important implications for the type of strategies chosen in the development of efficacious vaccines for patients with metastatic melanoma. PMID- 9856822 TI - EGF stimulates transcription of CaN19 (S100A2) in HaCaT keratinocytes. AB - CaN19 (S100A2), a member of the S100 family of calcium-binding proteins, was originally isolated in a screen for tumor suppressor genes. Recent work from our laboratory suggests that CaN19 is likely to be an effector of the regenerative hyperplasia pathway of epidermal differentiation. As other work from our laboratory in a human skin organ culture model suggests that this response is mediated by activation of the epidermal growth factor (EGF) receptor and/or related receptors of the ErbB family, we asked whether CaN19 expression could be increased by organ culture and by EGF treatment of human keratinocytes. CaN19 was strongly induced after 24 h of organ culture, and its induction could be blocked by PD153035, a specific inhibitor of EGF receptor tyrosine kinase activity. EGF treatment of immortalized human keratinocytes (HaCaT cells) increased CaN19 mRNA levels by 4.5-fold within 8 h, and a corresponding increase in CaN19 protein was observed by western blotting. EGF treatment had no effect on the expression of five other members of the S100A gene cluster. As assessed by nuclear run-off assay, CaN19 transcription increased rapidly in response to EGF, reaching a maximum induction of 16-fold after 2 h. In contrast, EGF treatment had no detectable effects on the decay of CaN19 transcripts, which were long lived (t1/2 > 6 h) in the presence or absence of EGF. PD153035 also blocked CaN19 transcription and the accumulation of CaN19 mRNA and protein in HaCaT cells. These results demonstrate that EGF receptor activation selectively stimulates CaN19 gene expression at the transcriptional level in human keratinocytes, and support the hypothesis that CaN19 is an important mediator of regenerative epidermal hyperplasia. PMID- 9856823 TI - Cholesterol sulfate activates transcription of transglutaminase 1 gene in normal human keratinocytes. AB - Cholesterol sulfate and transglutaminase 1 are essential for the process of keratinization. Cholesterol sulfate is formed during keratinization and activates the eta isoform of protein kinase C. Transglutaminase 1 is a key enzyme for formation of the cornified envelope in terminally differentiated keratinocytes. In this study, we demonstrated that cholesterol sulfate acts as a transcriptional activator of the transglutaminase 1 gene in normal human keratinocytes. Growth of normal human keratinocytes was inhibited by cholesterol sulfate, but not by its parental cholesterol. Treatment of normal human keratinocytes with cholesterol sulfate induced activity of transglutaminase 1 in a dose- and time-dependent manner. Activation of transcription of transglutaminase 1 by cholesterol sulfate was demonstrated by northern blotting analysis, whereas that by cholesterol was not. In order to identify a cholesterol sulfate responsive region in the transglutaminase 1 gene, plasmids were constructed containing a luciferase reporter gene ligated to deletion fragments of the 5' upstream region of the tranglutaminase 1 gene and were transfected into normal human keratinocytes. Transfected cells were treated with cholesterol sulfate, the phorbol ester 12-O tetradecanoylphorbol-13-acetate and a high concentration of Ca2+. Our results indicate that the responsive element(s) for cholesterol sulfate and phorbol ester is located upstream of the human transglutaminase 1 gene at a position(s) between -819 and -549, whereas the responsive element for Ca2+ is located at a position between -79 and -49. PMID- 9856824 TI - Chimeric human epidermal reconstructs to study the role of melanocytes and keratinocytes in pigmentation and photoprotection. AB - Chimeric epidermal reconstructs made with Negroid melanocytes and Caucasoid keratinocytes (or vice versa) were studied before and after UVB irradiation to understand the respective roles of these cells in tanning and photoprotection, especially lipoperoxidation and enzymatic defences against free radicals. Using this approach, we have confirmed overall the theory of the epidermal melanin unit. We have also shown that melanocytes of poorly tanning Caucasoids, which have a comparatively higher content of unsaturated fatty acids in their cell membrane, are more prone to the peroxidative effects of UV light, and that keratinocytes participate in photoprotection via phototype-dependent antioxidant enzyme activities, especially for catalase. PMID- 9856825 TI - Keratinocyte-specific retinoid regulation of human cellular retinoic acid binding protein-II (hCRABPII) gene promoter requires an evolutionarily conserved DR1 retinoic acid-responsive element. AB - Transcription of the hCRABPII gene is retinoid inducible in human skin keratinocytes (KC) but, surprisingly, not in cultured cells. The promoter for the gene harbors three putative nuclear receptor binding sites: DR5, upstream of the transcription start site; DR1 (DR1d), distal to the site; and DR1 (DR1p), a proximal variant. DR1d, but not DR1p, is conserved between human and mouse. Although DR5 has been found to be a retinoid receptor target in COS-1 cells, the function of DR1 remains unknown. We examined the functions of these DR in retinoid regulation of the hCRABPII promoter in human KC. In reporter gene assays, no significant retinoid response was observed in the promoter in cultured KC; however, overexpression of retinoid receptor heterodimers RARgamma x RXRalpha restored the response. Gel supershift assays showed that endogenous RARgamma x RXRalpha levels are much lower in cultured KC than in skin in vivo. Ligand binding assays showed that cultured KC contain only one-third of the level of retinoic acid receptor (RAR) and one-eighth of the level of retinoid X receptor found in KC in skin. Deletion of the DR1d or DR5 sites reduced retinoid-induced promoter activity by 63% and 27%, respectively. Isolated DR1d and DR5 sites, but not DR1p, efficiently bound RARgamma-RXRalpha and conferred RAR-selective retinoid responsiveness on a heterologous promoter. These data indicate that: (i) the previously reported lack of retinoid regulation of endogenous hCRABPII gene transcription in cultured KC is likely due to insufficient levels of RARgamma x RXRalpha, but not their cofactors; (ii) the conserved DR1d site is the major functional target in RARgamma x RXRalpha regulation of hCRABPII in KC; (iii) the DR1p site is nonfunctional due to its lack of affinity for RARgamma x RXRalpha, although its half-sites share high sequence homology with the consensus retinoid receptor-binding half-site. PMID- 9856826 TI - Differential expression of peroxisome proliferator-activated receptor subtypes during the differentiation of human keratinocytes. AB - The expression of mRNA encoding peroxisome proliferator-activated receptor (PPAR) subtypes in human keratinocytes was determined by semiquantitative reverse transcriptase-polymerase chain reaction. When normal human keratinocytes were induced to differentiate by shifting the culture medium to high Ca2+ concentration, the expression of PPAR-alpha and -gamma mRNA was increased, whereas that of PPAR-delta remained unchanged. At the protein level, the expression of PPAR in cultured human keratinocytes was demonstrated by a DNA mobility shift assay and the functionality of the receptor subtypes was assessed by transactivation experiments. In epidermis reconstructed in vitro, the level of PPAR-alpha and -gamma mRNA was also associated with keratinocyte differentiation. In lesional compared with nonlesional psoriatic epidermis, the expression of PPAR alpha and -gamma mRNA was reduced, indicating that these two subtypes are tightly linked to the epidermal differentiation process. PMID- 9856827 TI - PTHrP regulates epidermal differentiation in adult mice. AB - Emerging evidence suggests that parathyroid hormone-related peptide (PTHrP) serves as a regulator of the development and/or differentiation of a number of organs, including endochondral bone, the tooth, and the mammary gland. Although disruption of the PTHrP gene by homologous recombination results in a lethal chondrodystrophy, PTHrP-knockout mice that have been rescued by the transgenic replacement of the peptide in cartilage display abnormalities in ectodermally derived structures including the skin. At 6-8 wk of age, these rescued PTHrP knockout mice displayed a markedly thinned epidermis and striking hyperkeratosis, hypoplastic sebaceous glands, and a fibrotic dermis. In contrast, transgenic mice that overexpress PTHrP by virtue of the human keratin-14 promoter displayed a thickened ventral epidermis with marked acanthosis and papillomatosis, hyperplastic sebaceous glands, and a cellular dermis. The absence of PTHrP appeared to result in the reduction of the basal keratinocyte compartment and premature acquisition of suprabasal and granular differentiation markers, whereas overexpression of the peptide generated reciprocal findings. No difference in the epidermal proliferation rate was found in PTHrP-null skin and although an increase was observed in keratin 14-PTHrP transgenic animals, their epidermis did not express the hyperplasia marker K6. Finally, the replacement of PTHrP in the basal keratinocytes of rescued PTHrP-knockout mice under the direction of the keratin 14 promoter reversed the abnormalities seen in PTHrP-null skin. These findings suggest that PTHrP regulates the rate of keratinocyte differentiation in the skin of adult mice. PMID- 9856828 TI - Water and protein structure in photoaged and chronically aged skin. AB - Changes in the structural proteins and hydration during aging is responsible for altered skin morphologic and mechanical properties manifested as wrinkling, sagging, loss of elasticity, or apparent dryness. To gain insight into the age related alterations in protein conformation and water structure, we obtained Raman spectra from the sun-protected buttock skin representing chronologic aging and the sun-exposed forearm skin representing combined effects of photoaging and chronologic aging. Ten aged individuals (five men, five women; age range 74-87) and 10 control young individuals (five men, five women; age range 22-29) entered the study. In the photoaged forearm skin the positions of protein-specific amide I, amide III, and CH stretching bands were shifted, suggesting increased protein folding. In contrast, major changes were seen only in the amide I peak in chronologically aged skin. The intensity of the 3250 cm(-1) OH stretching band was increased in photoaged skin (but not in chronologically aged skin) indicating an increased water content. R(v) representation of the low-frequency region of Raman spectra was applied to determine water structure. In the young skin and chronologically aged skin water was mostly present in the bound form. In the photoaged skin, however, an increase in intensity at 180 cm(-1) was noted, which reflects an increase in the not-protein bound water (tetrahedron water clusters). In conclusion, it seems that proteins in the photoaged skin are more compact and interact with water to limited degree. Impairment in protein hydration may add to the understanding of ultrastructural, mechanical, and biochemical changes in structural proteins in the aged skin. PMID- 9856829 TI - Reduction in number and morphologic alterations of Langerhans cells after UVB radiation in vivo are accompanied by an influx of monocytoid cells into the epidermis. AB - Acute, low-dose ultraviolet B (UVB) radiation impairs contact hypersensitivity induction in mice by a mechanism due at least in part to Langerhans cells alterations. To better define the effects of UVB on Langerhans cells, we have compared the action of this agent on the skin of intact mice and in skin explants incubated in vitro up to 24 h. Using immunofluorescence, we detected a reduction in the length of the dendrites of Langerhans cells and a significant reduction in the number of Ia-positive Langerhans cells per unit area within 2 h of UVB; these changes reversed within 24 h in vivo, but not in vitro. By electron microscopy, the number of dendritic cells per 100 basal keratinocytes increased in vivo, but decreased in vitro by 2 h after UVB, a discordance that was significant. On the contrary, the number of dendrite profiles per dendritic cell body decreased significantly 2 h after UVB, both in vivo and in vitro. Many epidermal dendritic cells, 2 h after UVB in vivo, were deficient in cytoplasmic organelles, whereas the few cells that remained after UVB in vitro retained their Birbeck granules, and displayed many, dilated cytoplasmic vesicles. We interpret these data to mean that low doses of UVB radiation destroy the functional and morphologic integrity of epidermal Langerhans cells, and that these cells are rapidly replaced by precursor cells that mature in situ into normal-appearing Langerhans cells. PMID- 9856830 TI - Wound fluid from venous leg ulcers degrades plasminogen and reduces plasmin generation by keratinocytes. AB - Plasminogen, the pro-enzyme of plasmin, aids various processes essential for normal, acute wound healing, such as fibrinolysis and cell migration. We have investigated if plasminogen is available to perform these functions in chronic wounds such as venous leg ulcers. We report that plasminogen is degraded by fluid from venous leg ulcers to a number of fragments, including kringle domains 1-3, an angiostatin-related protein. The enzyme responsible was inhibited by the serine protease inhibitor phenyl-methylsulfonyl fluoride, but was not inhibited by alpha1-anti-trypsin, an inhibitor of neutrophil elastase, by alpha2-anti plasmin, an inhibitor of plasmin, or by the matrix metalloprotease inhibitor 1,10 phenanthroline. Plasminogen degraded by wound fluid was a weaker substrate than intact plasminogen for plasmin generation by the keratinocyte cell line HaCaT. These results suggest that serine protease activity in leg ulcer fluid degrades plasminogen and support the hypothesis that keratinocyte migration may be impaired in leg ulcers because of a reduced availability of intact plasminogen for plasmin generation. PMID- 9856831 TI - Induction of bone morphogenetic protein-6 in skin wounds. Delayed reepitheliazation and scar formation in BMP-6 overexpressing transgenic mice. AB - Growth factors of the transforming growth factor-beta superfamily are involved in cutaneous wound healing. In this study we analyze the expression of the bone morphogenetic protein-6 (BMP-6) gene, a transforming growth factor-beta related gene, in skin wounds. In normal mouse skin high levels of BMP-6 mRNA and protein are expressed by postmitotic keratinocytes of stratified epidermis until day 6 after birth. BMP-6 expression is strongly reduced in adult epidermis with diminished mitotic activity. After skin injury we found large induction of BMP-6 specific RNA and protein in keratinocytes at the wound edge and keratinocytes of the newly formed epithelium as well as in fibroblast shaped cells in the wound bed. BMP-6-specific RNA was induced within 24 h after injury, whereas significant upregulation of BMP-6 on the protein level was detected only 2-3 d after injury. Protein was confined to outermost suprabasal epidermal layers, whereas BMP-6 specific RNA was distributed throughout all epidermal layers including basal keratinocytes and the leading edge of the migrating keratinocytes. We also detected high levels of BMP-6-specific RNA and protein in chronic human wounds of different etiology. In contrast to the overall distribution pattern of BMP-6 specific RNA, the protein was not detected in keratinocytes directly bordering the wound. In order to test the influence of BMP-6 abundance on the progress of wound healing, we analyzed the wound response of transgenic mice overexpressing BMP-6 in the epidermis. In these mice, reepitheliazation of skin wounds was significantly delayed, suggesting that strict spatial and temporal regulation of BMP-6 expression is necessary not only for formation but also for reestablishment of a fully differentiated epidermis. PMID- 9856832 TI - Membrane-type matrix metalloproteinases in human dermal microvascular endothelial cells: expression and morphogenetic correlation. AB - Membrane-type matrix metalloproteinases (MT-MMP) activate the zymogen form of MMP 2/Gelatinase A on cell surfaces and are expressed in invasive tumors. We sought to identify and characterize MT-MMP in a non-malignant cell type that undergoes a physiologic and reversible invasive phenotype during angiogenesis. Human dermal microvascular endothelial cells (HDMEC) were isolated from neonatal tissue and purified by anti-CD31 (PECAM) affinity beads. MT-MMP-1 and -3 transcripts were amplified by reverse transcriptase-polymerase chain reaction and northern blots showed a single 4.5 kB mRNA for MT-MMP-1 that was modulated by angiogenic factors and phorbol ester. Immunoblotting of reduced cellular extracts with different MT MMP-1 antibodies showed the presence of the 63-65 kDa and 57-60 kDa forms, as well as additional forms at lower molecular weights. HDMEC membranes extracted with Triton X114 were incubated with gelatin-sepharose purified MMP-2 and MMP-9 to show activation of proenzymes. Pre-incubation of HDMEC with anti-MT-MMP-1 antibodies decreased proMMP-2 conversion activity only. The movement of HDMEC and the formation of tubule-like structures in three-dimensional collagen gels was markedly delayed by preincubation with the same anti-MT-MMP-1 antibodies. These results demonstrate the presence of MT-MMP in cutaneous microvascular cells in vitro. Modulation of these cell surface proteinases by angiogenic factors, demonstration of multiple processed forms, and specific attenuation of HDMEC morphogenetic patterns in three-dimensional collagen gels implicate their potential roles in the formation of new blood vessels in the skin. PMID- 9856833 TI - Hepatocyte growth factor/scatter factor (HGF/SF) induces vascular permeability factor (VPF/VEGF) expression by cultured keratinocytes. AB - Skin expression of the endothelial cell-specific vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) as an outstanding mediator of physiologic and pathologic angiogenesis has been previously demonstrated to be subject to regulation by distinct stimuli. We explored whether the multifunctional hepatocyte growth factor/scatter factor (HGF/SF) may mediate its angiogenic properties in part through paracrine induction of cutaneous VPF/VEGF synthesis. In these studies, we demonstrate that HGF/SF functions as a potent inducer of VPF/VEGF expression by human epidermal keratinocytes and by different epithelial-derived cells in vitro. VPF/VEGF mRNA and protein expression are regulated by HGF/SF in both a concentration- and a time-dependent fashion. Examination of mRNA half-lives does not reveal an increase in VPF/VEGF mRNA stability after HGF/SF stimulation. Thus, HGF/SF-induced VPF/VEGF mRNA expression appears to be largely dependent on enhanced gene transcription. In analyses of transiently transfected 5'-deletional reporter gene constructs, we identified a GC-rich VPF/VEGF promoter element that conveys transcriptional activation in response to HGF/SF. This sequence, located between nucleotides -88 and -70, is critical for both constitutive and HGF/SF-induced transcriptional activity. Together, our observations support a model in which HGF/SF mediates angiogenic properties in part through paracrine induction of VPF/VEGF synthesis by keratinocytes. In addition to cutaneous inflammation and wound healing, our findings have potential significance for vascular hyperpermeability and angiogenesis in tumor growth. PMID- 9856834 TI - An experimental model for interpreting percutaneous penetration of oligonucleotides that incorporates the role of keratinocytes. AB - Oligonucleotides have been extensively studied for their potential as therapeutic agents. Phosphorothioate oligonucleotides have been demonstrated to be particularly useful due to their stability against nucleases, their ability to be internalized by many cell types, and the ease with which they hybridize with target mRNA. These compounds have previously been delivered across the skin with the aid of iontophoresis. During transdermal delivery, the first viable cells exposed to the oligonucleotides are the keratinocytes. The purpose of this study was to determine the relationship between internalization of these compounds by keratinocytes and their transport across the skin. The in vitro uptake of 15 different fluorescently labeled phosphorothioate oligonucleotides into human keratinocytes was quantitatively measured with a fluorometer. Photomicrographs of keratinocytes indicate diffuse cytoplasmic and nuclear localization. The ability of these molecules to enter cells was linearly related to size. Cellular uptake data were inversely correlated with previously reported steady-state transport levels of oligonucleotides that had been transdermally delivered by iontophoresis across hairless mouse skin. Oligonucleotides that readily entered keratinocytes had a decreased ability to penetrate skin under iontophoretic conditions. The results indicate that oligonucleotide sequences may be designed for treating skin diseases (high uptake, low transport) or systemic disorders (low uptake, high transport). PMID- 9856835 TI - Different frequency of gene targeting events by the RNA-DNA oligonucleotide among epithelial cells. AB - A unique hybrid oligonucleotide composed of both RNA and DNA has been shown to correct a point mutation in a site-specific and inheritable manner in extrachromosomal and chromosomal targets. In order to develop new gene therapeutics for skin, we tested two oligonucleotides that were shown to create a point mutation in alkaline phosphatase and beta-globin genes in several epithelial cell types. Highly transformed epithelial cells (HeLa) exhibited a conversion frequency of 5% by both RNA-DNA oligonucleotides. In comparison, other immortalized epithelial cells (HaCaT) or human primary keratinocytes did not show any detectable level of gene conversion by the restriction fragment length polymorphism analysis, indicating less than 1% conversion frequency. The concentration of the oligonucleotide in the nuclei of HeLa cells was similar to that of HaCaT or human primary keratinocytes measured by a radiolabeled or a fluorescein-conjugated oligonucleotide. Moreover, the RNA-DNA oligonucleotide exhibited a prolonged stability in the nucleus. Thus, neither uptake nor nuclear stability of the oligonucleotide appears to be a limiting factor in gene targeting events under our experimental conditions. These results indicate that the frequency of gene targeting varies among different cells, suggesting that cellular recombination and DNA repair activities may be important. PMID- 9856837 TI - Epidermal cytokines IL-1beta, TNF-alpha, and IL-12 in patients with atopic dermatitis: response to application of house dust mite antigens. AB - Epidermal cytokines such as interleukin (IL)-1beta, tumor necrosis factor-alpha, and IL-12 have been described to play a crucial role in the induction and elicitation phase of allergic contact dermatitis upon exposure to haptens. In this study we asked whether these cytokines may also play a role in the epidermis of patients with atopic dermatitis after the application of house dust mite antigens (HDM) to their skin. Epidermal samples were collected by scraping healthy appearing skin of atopic patients and healthy individuals 8 h after the application of an extract of HDM. Sodium lauryl sulfate and saline served as controls. Reverse transcriptase-polymerase chain reaction was performed for IL 1beta, tumor necrosis factor-alpha, IL-12 p35, and IL-12 p40. Exposure to HDM led to a significant upregulation of mRNA of these cytokines in atopic patients only. Whereas IL-1beta and tumor necrosis factor-alpha also showed an upregulation in part of these patients after exposure to the irritant sodium lauryl sulfate, IL 12 p40 mRNA was exclusively enhanced by the application of the allergen. In contrast to IL-12 p40, IL-12 p35 mRNA was not detectable in significant amounts. Interestingly, also in untreated, normal appearing skin of atopic individuals (n = 16), the levels of these cytokines were higher than in normal individuals (n = 8), possibly explaining the increased skin irritability of atopic individuals. Finally, comparing epidermal cytokines in the skin of patients who developed a positive allergen patch test to those who stayed negative, suggests that only expression of IL-1beta mRNA may be a predictive marker for the development of a positive patch test reaction to HDM. PMID- 9856836 TI - Molecular characterization of an autoallergen, Hom s 1, identified by serum IgE from atopic dermatitis patients. AB - Atopy is a genetically determined disorder that affects 10%-20% of the population. Many symptoms of patients with atopy (allergic rhinitis, conjunctivitis, asthma, and anaphylaxis) result from events occurring after crosslinking of cell-bound IgE by per se innocuous environmental antigens. The frequently raised hypothesis that autosensitization can also be a pathogenetic factor in atopy, gained support by our recent demonstration of IgE antibodies against human proteins in atopic dermatitis patients. To unravel the molecular nature of IgE-defined autoantigens, we used serum IgE from atopic dermatitis patients to screen a human epithelial cDNA expression library. One of the cDNA encoding IgE-reactive products contained 1501 bp of a 2274 bp open-reading frame finally identified by sequence analysis of two additional cDNA clones resulting from oligonucleotide screening. The IgE-defined autoantigen, designated Hom s 1, exhibited an almost complete sequence identity with a recently described antigen recognized by cytotoxic T cells of a squamous cell carcinoma patient. Purified recombinant Hom s 1 specifically bound IgE from patients with severe atopy. When used as immunogen in rabbits, recombinant Hom s 1 gave rise to an anti-serum that reacted with a cytoplasmic protein exhibiting a broad cellular and tissue reactivity (skin, lung >> gastrointestinal tract >> muscle, brain) and identified a 55 kDa protein in blotted serum IgE preparations. The attractive possibility remains that the Hom s 1-triggered IgE response contributes to the events resulting in allergic tissue inflammation. If so, the respective recombinant molecule may serve as a paradigmatic tool for the diagnosis and treatment of patients with "intrinsic" atopy. PMID- 9856838 TI - Neonatal lupus erythematosus: maternal IgG antibodies bind to a recombinant NH2 terminal fusion protein encoded by human alpha-fodrin cDNA. AB - IgG antibodies to a cleavage product of alpha-fodrin (120 kDa alpha-fodrin) have recently been identified as organ-specific autoantibodies in primary Sjogren's syndrome. In this study, we examined seroreactivity of mothers and infants with neonatal lupus erythematosus (NLE) to a recombinant NH2-terminal protein (120 kDa alpha-fodrin) of human alpha-fodrin. Serum samples were collected during the perinatal period in seven pregnancies of five mothers delivering offspring with NLE. Anti-120 kDa alpha-fodrin antibodies were identified by immunoblotting in six of seven perinatal maternal sera of offspring with NLE: one of two congenital heart block offspring and all five offspring with cutaneous NLE. These antibodies were placentally transmitted to infants. One of the five mothers had primary Sjogren's syndrome, and four were asymptomatic. One asymptomatic mother did not demonstrate anti-120 kDa alpha-fodrin activity at the time of the first delivery of a congenital heart block infant, but was found to be positive at the time of subsequent delivery of a second child with cutaneous NLE. We propose that maternal antibodies to 120 kDa alpha-fodrin may be an additional serologic marker for the risk of NLE in anti-Ro/SS-A positive women. PMID- 9856839 TI - Stimulation of collagen synthesis by the anabolic steroid stanozolol. AB - There is evidence that anabolic steroids, which are derived from testosterone and have markedly less androgenic activity, promote tissue growth and enhance tissue repair; however, the mechanisms involved in their anabolic activities remain unclear. In this report, we measured the effect of the anabolic steroid stanozolol on cell replication and collagen synthesis in cultures of adult human dermal fibroblasts. Stanozolol (0.625-5 microg per ml) had no effect on fibroblast replication and cell viability (p = 0.764) but enhanced collagen synthesis (p < 0.01) in a dose-dependent manner (r = 0.907). Stanozolol also increased (by 2-fold) the mRNA levels of alpha1 (I) and alpha1 (III) procollagen and, to a similar extent, upregulated transforming growth factor-beta1 (TGF beta1) mRNA and peptide levels (p < 0.001). There was no stimulation of collagen synthesis by testosterone. The stimulatory effects of stanozolol on collagen synthesis were blocked by a TGF-beta1 anti-sense oligonucleotide, by antibodies to TGF-beta, and in dermal fibroblast cultures derived from TGF-beta1 knockout mice. We conclude that collagen synthesis is increased by the anabolic steroid stanozolol and that, for the most part, this effect is due to TGF-beta1. These findings point to a novel mechanism of action of anabolic steroids. PMID- 9856840 TI - Acute barrier perturbation abolishes the Ca2+ and K+ gradients in murine epidermis: quantitative measurement using PIXE. AB - Epidermal permeability barrier homeostasis requires the delivery of lipids and hydrolytic enzymes by lamellar body exocytosis from the uppermost granular cells, a process that is upregulated following barrier disruption. As lamellar body secretion is controlled by ionic concentrations, especially Ca2+ and K+, we used a quantitative technique, microbeam proton-induced X-ray emission, to measure Ca2+, K+, Cl-, and P concentrations before and after acute barrier perturbation by acetone applications. We found a steep gradient of Ca2+ in normal tissue, peaking in the outer stratum granulosum, which disappeared after barrier disruption, and partially reformed as the barrier recovered. A similar gradient, peaking somewhat lower in the epidermis (i.e., at the stratum granulosum-stratum corneum interface), was found for K+. Epidermal concentrations of K+ also decreased after barrier abrogation, although to a lesser extent than Ca2+. In contrast, P and Cl- demonstrated distribution gradients at baseline, which remained unchanged after barrier disruption. These studies quantitate the levels of Ca2+, K+, Cl-, and P within specific epidermal cell layers at baseline, and in relation to changes in permeability barrier integrity. Ca2+ and K+, but not Cl- or P, decrease after barrier disruption, consistent with these two ion's role in barrier repair. PMID- 9856841 TI - CDKN2a/p16INK4a mutations and lack of p19ARF involvement in familial melanoma kindreds. AB - Germline mutations in the cyclin-dependent kinase inhibitor 2a (CDKN2a) gene, which maps to the 9p21 chromosomal region and encodes the cyclin-dependent kinase inhibitor p16INK4a, have been detected in a proportion of familial melanoma kindreds, suggesting that it is the putative 9p21-linked melanoma susceptibility gene. The p19ARF transcript, an alternative spliced form of the CDKN2a gene, has recently been shown to inhibit, like the p16INK4a protein, cell cycle progression, raising the possibility that it might constitute an additional melanoma tumor suppressor gene at the 9p21 locus. To determine the contribution of these candidate genes to familial melanoma genetic predisposition, we screened 10 such kindreds for germline mutations in the p16INK4a and p19ARF genes. Four independent germline missense mutations, mapping in exon 1alpha (Gly23Asp; Arg24Pro) and exon 2 (Asn71Ileu; Pro114Leu) of the CDKN2a gene, were identified. Two previously described polymorphisms were also detected, Ala148Thr in exon 2 and a base change in the 3' untranslated region of exon 3. No disease-associated mutations in exon 1beta of the p19ARF gene were found. Our data support the hypothesis that the CDKN2a is a melanoma susceptibility gene in familial melanoma, whereas the p19ARF gene does not seem to play a significant role. PMID- 9856842 TI - Mutations in keratin K9 in kindreds with epidermolytic palmoplantar keratoderma and epidemiology in Northern Ireland. AB - Epidermolytic palmoplantar keratoderma (EPPK, MIM #144200) is an autosomal dominant disorder in which hyperkeratosis confined to the palms and soles is characterized histologically by cytolysis of suprabasal keratinocytes. Mutations in the keratin 9 gene (KRT9), a type 1 keratin expressed exclusively in the suprabasal keratinocytes of palmoplantar epidermis, have previously been demonstrated in this disorder. Here, we have studied four Northern Irish kindreds presenting with EPPK. By direct sequencing of polymerase chain reaction products, heterozygous missense mutations in exon 1 of KRT9 were detected in all the families. These included a novel mutation M156T; as well as M156V in two kindreds; and R162Q in the remaining family. All mutations were confirmed by reverse strand sequencing and restriction enzyme analysis. The point prevalence of EPPK in Northern Ireland was found to be 4.4 per 100,000. To date, all reported EPPK mutations occur in the helix initiation motif at the start of the central coiled-coil rod domain of K9. PMID- 9856843 TI - Novel and de novo glycine substitution mutations in the type VII collagen gene (COL7A1) in dystrophic epidermolysis bullosa: implications for genetic counseling. AB - The dystrophic forms of epidermolysis bullosa (DEB) are due to mutations in the type VII collagen gene (COL7A1). In dominant DEB, a characteristic genetic lesion is a glycine substitution mutation within the collagenous domain of the protein. In this study, we have examined the molecular basis of six new families in which the proband has clinical features and/or ultrastructural findings consistent with DEB. The results revealed a glycine substitution mutation in all six families, four of which are novel and previously unpublished. In three families with clinically unaffected parents, de novo mutations G2043R and G2040V were found. These results emphasize the predominance of glycine substitution mutations in dominant DEB, and indicate that in some cases the phenotype is due to de novo dominant mutations. PMID- 9856844 TI - Transient bullous dermolysis of the newborn associated with compound heterozygosity for recessive and dominant COL7A1 mutations. AB - The neonatal skin blistering disorder transient bullous dermolysis of the newborn (TBDN) heals spontaneously or improves dramatically within the first months and years of life. TBDN is characterized by subepidermal blisters, reduced or abnormal anchoring fibrils at the dermo-epidermal junction, and electron-dense inclusions in keratinocytes. These features are partly similar to those in dystrophic epidermolysis bullosa, which is caused by defects in COL7A1 gene encoding collagen VII, the main anchoring fibril protein. TBDN has been grouped separately from dystrophic epidermolysis bullosa based on the pronounced morphologic features and the self-limiting course of the disorder; however, it remains unclear whether it represents a distinct clinical entity with a single etiology. We now report a TBDN patient who is compound heterozygous for a recessive and a dominant glycine substitution mutation in COL7A1. Two point mutations caused amino acid substitutions G1519D and G2251E in the triple helical domain of collagen VII. In the heterozygous state G1519D was silent, and G2251E led to nail dystrophy, but not to skin blistering. In the proband, compound heterozygosity for the mutations caused massive, transitory retention of collagen VII in the epidermis, its reduced deposition at the basement membrane zone, and extensive dermo-epidermal separation at birth. Accordingly, TBDN keratinocytes in vitro accumulated collagen VII intracellularly in the rough endoplasmic reticulum. PMID- 9856845 TI - A novel helix termination mutation in keratin 10 in annular epidermolytic ichthyosis, a variant of bullous congenital ichthyosiform erythroderma. AB - Annular epidermolytic ichthyosis is a distinct phenotypic variant of bullous congenital ichthyosiform erythroderma that has recently been described in two separate kindreds. Individuals with this variant present with bullous ichthyosis in early childhood and hyperkeratotic lichenified plaques in the flexural areas and extensor surfaces at later ages. Characteristically, they also develop intermittent bouts of annular and polycyclic, erythematous, scaly plaques on the trunk and proximal extremities. We now describe a third kindred with annular epidermolytic ichthyosis. Molecular analysis of this family revealed a novel mutation resulting in an isoleucine to threonine substitution at residue 107 (codon 446) within the highly conserved helix termination motif at the end of the rod domain of keratin 10. PMID- 9856846 TI - An atypical form of bullous congenital ichthyosiform erythroderma is caused by a mutation in the L12 linker region of keratin 1. AB - Defective keratins are the cause of a number of hereditary disorders of the epidermis and other epithelia. The disease-causing mutations in keratins are clustered in the rod domain, and mutations in the helix boundary peptides cause the most severe forms of epidermal fragility syndromes. Siemens described a family with an atypical, mild form of bullous congenital ichthyosiform erythroderma. Linkage analysis in this family indicated that a defective type II keratin might be the underlying cause, keratins K1 and K2e being the best candidates. A substitution of valine for aspartic acid was detected at position 340 (D340V) in the L12 region of the K1 polypeptide. The mutation was found to cosegregate with the disorder in the family. Herewith, a genotype-phenotype correlation is shown for bullous congenital ichthyosiform erythroderma comparable with that described for epidermolysis bullosa simplex. PMID- 9856847 TI - Identification of activating c-kit mutations in adult-, but not in childhood onset indolent mastocytosis: a possible explanation for divergent clinical behavior. AB - Mastocytosis represents a mast cell proliferative disease that generally runs a benign clinical course, with spontaneous remissions mostly by puberty in childhood-onset disease, although rare forms, particularly in adult-onset disease, can be associated with (pre)malignant hematologic disorders and very rarely present as mast cell leukemia or malignant mastocytosis. Reasons for this divergent clinical behavior of childhood- versus adult-onset disease are unknown. Recently, two activating mutations in the intracellular domain of the proto oncogene c-kit, which encodes a tyrosine kinase receptor for the mast cell growth factor stem cell factor, have been detected in the human leukemic mast cell line HMC-1. We have therefore studied lesional skin biopsies from patients with adult- and childhood-onset indolent mastocytosis for the presence of these codon 560 and 816 mutations. C-kit coding DNA sequences were amplified and analyzed by mutation specific restriction analyses, and mutated polymerase chain reaction products were additionally cloned and sequenced. The codon 816 mutation was found in all six samples from adult patients, but not in any of the 11 specimens from children. In addition, the codon 560 mutation could be demonstrated for the first time in indolent mastocytosis, namely in two of four specimens from adult patients, but not in those from two children. These data thus provide a possible explanation for the divergent clinical behavior of adult- versus childhood-onset indolent mastocytosis, with the first being associated with an activating mutation, possibly as part of a neoplastic process, and the latter representing most likely a reactive process of an as yet unknown pathogenesis. PMID- 9856848 TI - Langerhans cells express matrix metalloproteinase-9 in the human epidermis. PMID- 9856849 TI - Novel premature termination codon mutations in the laminin gamma2-chain gene (LAMC2) in Herlitz junctional epidermolysis bullosa. PMID- 9856850 TI - Treatment of psoriasis with interleukin-10. PMID- 9856851 TI - Envoplakin and periplakin are components of the paraneoplastic pemphigus antigen complex. PMID- 9856852 TI - Combination of a novel frameshift mutation (1929delCA) and a recurrent nonsense mutation (W610X) of the LAMB3 gene in a Japanese patient with Herlitz junctional epidermolysis bullosa, and their application for prenatal testing. PMID- 9856853 TI - Are His63Asp or Cys282Tyr HFE mutations associated with porphyria cutanea tarda? Data of patients from central and southern Italy. PMID- 9856854 TI - Do primary cutaneous non-T non-B CD4+CD56+ lymphomas belong to the myelo monocytic lineage? PMID- 9856855 TI - Heterozygosity for premature termination codon mutations in LAMB3 in siblings with non-lethal junctional epidermolysis bullosa. PMID- 9856856 TI - A novel point mutation of the EDA gene in a Japanese family with anhidrotic ectodermal dysplasia. PMID- 9856857 TI - Evidence for involvement of Bax and p53, but not caspases, in radiation-induced cell death of cultured postnatal hippocampal neurons. AB - Bax (a death-promoting member of the bcl-2 gene family), the tumor suppressor gene product p53, and the ICE/ced-3-related proteases (caspases) have all been implicated in programmed cell death in a wide variety of cell types. However, their roles in radiation-induced neuronal cell death are poorly understood. In order to further elucidate the molecular mechanisms underlying radiation-induced neuronal cell death, we have examined the ability of ionizing radiation to induce cell death in primary cultured hippocampal neurons obtained from wild-type, p53 deficient and Bax-deficient newborn mice. Survival in neuronal cultures derived from wild-type mice decreased in a dose-dependent manner 24 hr after a single 10 Gy to 30 Gy dose of ionizing radiation. In contrast, neuronal survival in irradiated cultures derived from p53-deficient or Bax-deficient mice was equivalent to that observed in control, nonirradiated cultures. Western blot analyses indicated that neuronal p53 protein levels increased after irradiation in wild-type cells. However, Bax protein levels did not change, indicating that other mechanisms exist for regulating Bax activity. Adenovirus-mediated overexpression of p53 also caused neuronal cell death without increasing Bax protein levels. Irradiation resulted in a significant induction in caspase activity, as measured by increased cleavage of fluorogenic caspase substrates. However, specific inhibitors of caspase activity (zVAD-fmk, zDEVD-fmk and BAF) failed to protect postnatal hippocampal neurons from radiation-induced cell death. Staurosporine (a potent inducer of apoptosis in many cell types) effectively induced neuronal cell death in wild-type, p53-deficient and Bax deficient hippocampal neurons, indicating that all were competent to undergo programmed cell death. These results demonstrate that both p53 and Bax are necessary for radiation-induced cell death in postnatal cultured hippocampal neurons. The fact that cell death occurred despite caspase inhibition suggests that radiation-induced neuronal cell death may occur in a caspase-independent manner. PMID- 9856858 TI - Hippocampal AMPA and NMDA mRNA levels correlate with aberrant fascia dentata mossy fiber sprouting in the pilocarpine model of spontaneous limbic epilepsy. AB - There is considerable controversy whether aberrant fascia dentata (FD) mossy fiber sprouting is an epiphenomena related to neuronal loss or a pathologic abnormality responsible for spontaneous limbic seizures. If mossy fiber sprouting contributes to seizures, then reorganized axon circuits should alter postsynaptic glutamate receptor properties. In the pilocarpine-status rat model, this study determined if changes in alpha amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) and n-methyl-D-aspartic acid (NMDA) receptor subunit mRNA levels correlated with mossy fiber sprouting. Sprague-Dawley rats were injected with pilocarpine (320 mg/kg; i.p.) and maintained in status epilepticus for 6 to 8 hours (pilocarpine-status). Rats were killed during the: (1) latent phase after neuronal loss but before spontaneous limbic seizures (day 11 poststatus; n = 7); (2) early seizure phase after their first seizures (day 25; n = 7); and (3) chronic seizure phase after many seizures (day 85; n = 9). Hippocampi were studied for neuron counts, inner molecular layer (IML) neo-Timm's staining, and GluR1-3 and NMDAR1-2b mRNA levels. Compared with controls, pilocarpine-status rats in the: (1) latent phase showed increased FD GluR3, NMDAR1, and NMDAR2b; greater CA4 and CA1 NMDAR1; and decreased subiculum GluR1 hybridization densities; (2) early seizure phase showed increased FD GluR3, increased CA1 NMDAR1, and decreased subiculum NMDAR2b densities; and (3) chronic seizure phase showed increased FD GluR2; increased FD and CA4 GluR3; decreased CA1 GluR2; and decreased subiculum GluR1, GluR2, NMDAR1, and NMDAR2b levels. In multivariate analyses, greater IML neo-Timm's staining: (1) positively correlated with FD GluR3 and NMDAR1 and (2) negatively correlated with CA1 and subiculum GluR1 and GluR2 mRNA levels. These results indicate that: (1) hippocampal AMPA and NMDA receptor subunit mRNA levels changed as rats progressed from the latent to chronic seizure phase and (2) certain subunit alterations correlated with mossy fiber sprouting. Our findings support the hypothesis that aberrant axon circuitry alters postsynaptic hippocampal glutamate receptor subunit stoichiometry; this may contribute to limbic epileptogenesis. PMID- 9856859 TI - NT-3-mediated TrkC receptor activation promotes proliferation and cell survival of rodent progenitor oligodendrocyte cells in vitro and in vivo. AB - We have previously described the expression of a functional full-length trkC transcript for neurotrophin-3 (NT-3) receptor in oligodendroglia (OL) cells (Kumar and de Vellis, 1996). To date, the role of NT-3 and its signal transduction cascade in OL remains poorly defined. We report that the NT-3 responsive population of cells in the OL lineage are the progenitor cells and that the addition of NT-3 results in the autophosphorylation of p145TrkC. Furthermore, NT-3-mediated activation of p21ras and mitogen-activated protein kinase (MAPK), extracellular signal-regulated protein kinase2 (ERK2), were also observed in the progenitor OL cells. These protein tyrosine kinase (PTK)-induced responses were sensitive to the presence of K252a, an inhibitor for tyrosine kinase. We have determined that NT-3 promotes progenitor OL cell commitment to enter into S-phase of cell cycle to initiate DNA synthesis, in a manner similar to platelet-derived growth factor-AA (PDGF-AA). NT-3 thus plays a role in cell proliferation when present alone, while augmenting the proliferation capacity of PDGF-AA as indicated by the nuclear binding activity of the transcription factor, E2F-1. Both the initiation and progression of mitotic events were confirmed by the expression of c-myc and cdc2 in the presence of NT-3, PDGF-AA or NT-3 plus PDGF-AA. A cell survival assay examining interleukin 1-beta-converting enzyme (ICE)-like protease-mediated cleavage of poly (ADP-ribose) polymerase (PARP) revealed an increase in OL progenitor cell death in the absence of NT-3 or PDGF AA. In corroboration with our in vitro studies, in vivo results show an increased expression of the progenitor OL cell marker, glycerol phosphate dehydrogenase (GPDH) within 48 hr following an intracranial injection of NT-3, PDGF-AA, or NT-3 plus PDGF-AA in PN4-5 rats. These novel findings suggest that PDGF-AA potentiates the OL progenitor cell's ability to enter into the S-phase of the cell cycle and that NT-3 can augment this activity. Furthermore, PDGF-AA and NT-3 can block ICE like protease-mediated PARP fragmentation in progenitor OL cells. These results provide important information which further delineates the signal transduction cascades and the role of NT-3 and PDGF-AA on OL progenitor cells. PMID- 9856860 TI - Permanent rescue of lesioned neonatal motoneurons and enhanced axonal regeneration by adenovirus-mediated expression of glial cell-line-derived neurotrophic factor. AB - Axotomy of peripheral nerves in neonatal rats induces motoneuron death that can be delayed but not arrested by the application of several neurotrophic factors (NFs) or adenoviral vectors carrying genes for NFs. We tested whether an adenoviral vector carrying the gene for glial cell-line-derived neurotrophic factor (Adv.RSV-GDNF) would prevent neonatal motoneuron death after facial nerve transection or crush. Nerve transection eliminates the pathway for axonal regeneration, while nerve crush preserves the pathway necessary for target reinnervation that may be required for the permanent rescue of motoneurons. Both types of injury cause substantial motoneuron death in neonatal animals. Adv.RSV GDNF or a control vector carrying the beta-galactosidase gene (Adv.RSV-betagal) was injected into facial muscles 2 days before the nerve was transected, or Adv.RSV-GDNF, Adv.RSV-betagal, Adv.d1312 (a vector lacking a transgene), or vehicle was injected into facial muscles immediately after nerve crush. Four weeks after nerve transection, few motoneurons survived after Adv.RSV-GDNF and Adv.RSV-betagal treatment (6.1% and 2.4%, respectively). Four weeks after nerve crush, 40% of the motoneurons survived after Adv.RSV-GDNF treatment but only 17% survived in control groups. By 20 weeks, 39% of the motoneurons of the Adv.RSV GDNF treatment groups survived but only 15-19% survived in controls. The numbers of myelinated axons of the buccal nerve branch of Adv.RSV-GDNF treatment groups were also higher than controls at 4 and 20 weeks (24% and 100% compared to 4.4 6.2% and 25-33% for Adv.RSV-GDNF and controls, respectively). By 20 weeks, Adv.RSV-GDNF-treated animals recovered 50% of the contralateral vibrissal function, while in controls only 5-11% of function was restored. PMID- 9856861 TI - Oxidant treatment causes a dose-dependent phenotype of apoptosis in cultured motoneurons. AB - Evidence is growing that reactive oxygen species (ROS), by-products of (normal) cellular aerobic metabolism, are involved in the pathogenesis of neurodegenerative diseases. One of these diseases is amyotrophic lateral sclerosis (ALS), in which motoneurons die, leading to paralysis and death. It remains uncertain whether ROS are the cause of (apoptotic) motoneuron death in ALS. To further understand the role of ROS in motoneuron death, we investigated the effects of ROS on isolated spinal rat motoneurons in culture. ROS were generated with a combination of iron(III) and ascorbate, or with hydrogen peroxide. Both toxic treatments resulted in a dose-dependent motoneuron death. Iron(III)/ascorbate toxicity was completely prevented with the hydrogen peroxide detoxifying enzyme catalase and partially prevented with the antioxidant vitamin E. SOD1, the enzyme that removes superoxide, did not protect against iron(III)/ascorbate toxicity. ROS treatment caused apoptotic motoneuron death: low doses of iron(III)/ ascorbate or hydrogen peroxide resulted in complete apoptosis ending in nuclear fragmentation, while high doses of ROS resulted in incomplete apoptosis (nuclear condensation). Thus, depending on the dose of ROS, the motoneurons complete the apoptotic pathway (low dose) or are stopped somewhere during this route (high dose). PMID- 9856862 TI - Pregnenolone sulfate modulates NMDA receptors, inducing and potentiating acute excitotoxicity in isolated retina. AB - Pregnenolone sulfate (PS) acts as a positive allosteric modulator of N-methyl-D aspartate (NMDA) receptor-mediated responses. In the retina, we previously observed that the synthesis of pregnenolone and PS increases after stimulation of NMDA receptors and blockade of the synthesis reduces retinal cell death. This study was carried out to explore in the isolated and intact retina the possible role of PS in NMDA-induced excitotoxicity. Lactate dehydrogenase (LDH) measurements and morphological analysis revealed that a 90-min exogenous application of PS at 0.1-500 microM concentrations potentiated NMDA-induced cell death and at 50-500 microM concentrations caused cytotoxicity. After 45 min, either NMDA or PS caused no significant LDH release; but their co-application resulted in a high degree of toxicity. In addition, we found that a mild NMDA insult developed into serious damage when even low PS concentrations (0.1-10 microM) were used. Toxicity-inducing and -potentiating effects were specific to PS modulatory action on NMDA receptors, in that they were blocked by 4-(3 phosphonopropyl)2-piperazinecarboxylic acid (CPP) and MK-801 but not by 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX), and neither dehydroepiandrosterone sulfate nor pregnenolone caused LDH release. Prevention of degenerative signs was seen in retinae pretreated with 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), a Cl- channel blocker, thus indicating a Na+/Cl--dependent acute mode of excitotoxic cell death responsible for PS toxicity. The positive interaction between the neurosteroid and NMDA receptors was further proved by a PS dose dependent increase in NMDA-induced stimulation of [3H] MK-801 binding to retinal membranes. The results suggest a crucial role of PS in retinal vulnerability and propose the toxicity-potentiating effects as an important key in linking NMDA induced endogenous synthesis to acute excitotoxicity. PMID- 9856863 TI - Beta-amyloid binds to p57NTR and activates NFkappaB in human neuroblastoma cells. AB - Amyloid beta peptide (Abeta), a proteolytic fragment of the amyloid precursor protein (APP), is a major component of the plaques found in the brain of Alzheimer's disease (AD) patients. These plaques are thought to cause the observed loss of cholinergic neurons in the basal forebrain of AD patients. In these neurons, particularly those of the nucleus basalis of Meynert, an up regulation of 75kD-neurotrophin receptor (p75NTR), a nonselective neurotrophin receptor belonging to the death receptor family, has been reported. p75NTR expression has been described to correlate with beta-amyloid sensitivity in vivo and in vitro, suggesting a possible role for p75NTR as a receptor for Abeta. Here we used a human neuroblastoma cell line to investigate the involvement of p75NTR in Abeta-induced cell death. Abeta peptides were found to bind to p75NTR resulting in activation of NFKB in a time- and dose-dependent manner. Blocking the interaction of Abeta with p75NTR using NGF or inhibition of NFKB activation by curcumin or NFKB SN50 attenuated or abolished Abeta-induced apoptotic cell death. The present results suggest that p75NTR might be a death receptor for Abeta, thus being a possible drug target for treatment of AD. PMID- 9856864 TI - Docosahexaenoic acid decreases phospholipase A2 activity in the neurites/nerve growth cones of PC12 cells. AB - Docosahexaenoic acid (DHA) accumulates in nerve growth cones (NGC) during perinatal development and it is neuroprotective in ischemia. Because the phospholipases A2 (PLA2) are present in NGC and these enzymes function in both ischemia and long-term potentiation, the relationship between DHA and PLA2 was investigated in the NGC of nerve growth factor-differentiated PC12 cells. When PC12 cells were incubated with [3H]DHA, it primarily esterified in ethanolamine glycerolipids and concentrated initially in cell bodies with similar levels present in the neurite/nerve growth cone (N/NGC) fraction after 4 days. PLA2 activity in the N/NGC fraction was investigated using [14C]arachidonic acid labeled phosphatidylinositol ([14C-AA]PI) as substrate. Heat denaturation and pharmacological inhibition showed that much of the PLA2 activity was calcium independent and secretory rather than cytosolic. Supplementing the media with as little as 33 nM DHA significantly reduced PLA2 activity in the N/NGC fraction. PMID- 9856865 TI - Expression of glutamate receptors on cultured cerebral endothelial cells. AB - Activation of glutamate receptors has been shown to mediate a large number of neuronal processes such as long-term potentiation and ischemic damage. In addition to neurons and glia, glutamate receptors may occur on cerebral endothelial cells (CECs). The aim of the present study was to determine which glutamate receptors are expressed in CECs and to demonstrate the functional presence of such channels. By using reverse transcriptase-polymerase chain reaction, we showed that primary cultures of rat CECs express N-methyl-D aspartate (NMDA) receptors (NR1 subunit, which is necessary for the formation of functional NMDA receptors, and NR2A-C subunits), 2-amino-3-(3-hydroxy-5-methyl-4 isoxazolyl-propionate (AMPA) receptors (GLUR1-4 subunits), and metabotropic receptors (mGLUR). Exposure of the cultures to 2 mM glutamate, a well-established mediator of ischemic damage, for 30 min increased significantly the phosphorylation of calcium/calmodulin-dependent protein kinase II even after 10- and 60-min recovery times. This effect could be prevented by the NMDA blocker MK 801. The presence of multiple glutamate receptor types may confer a finely tuned responsiveness of the cerebral endothelium to glutamate in physiological and pathological conditions. PMID- 9856866 TI - ED2-positive perivascular phagocytes produce interleukin-1beta during delayed neuronal loss in the facial nucleus of the rat. AB - Injection of Fluoro-Gold (FG) into the whisker pad of rats yields stable retrograde labeling of facial motoneurons. Subsequent removal of 10 mm from all facial nerve branches permanently deprives the FG-labeled motoneurons from their targets and the motoneurons gradually die. Neuronal debris is phagocytized by two types of neuronophages: parenchymal microglia (monoclonal antibody [MAb] OX42 positive, MAb ED2-negative) and perivascular phagocytes (OX42-negative, ED2 positive). Because both types of neuronophages express major histocompatibility complex (MHC) class II glycoproteins (MAb OX6-positive), they are considered to be the potential antigen-presenting cells of the brain. To check this hypothesis, we tested whether both types of neuronophages also synthetize the co-stimulatory cytokine interleukin-1beta (IL-1beta) immunocytochemically visualized by MAbs SILK-5/6. Employing combined fluorescent visualization of antigens (OX6, ED2, and SILK-5/6) in sections containing fluorescent (FG-prelabeled) neuronophages, we found that, during slowly occurring neuronal loss, the vast majority of IL-1beta immunoreactive neuronophages were of perivascular (ED2-positive) origin. We concluded that, during delayed neuronal death "behind" an intact blood-brain barrier, the perivascular phagocytes were more likely to function as antigen presenting cells than the parenchymal microglia. PMID- 9856867 TI - Endothelial cell dysfunction in response to intracellular overexpression of amyloid precursor protein. AB - Previous reports have shown that exposure of vascular endothelial and smooth muscle cells to exogenous amyloid beta (Abeta) peptide results in cell damage and toxicity via oxidative injury. In this study we demonstrate that overexpression of the amyloid precursor protein (APP) is toxic to bovine aortic endothelial cells but not to bovine aortic smooth muscle cells. Intracellular coexpression of the free radical scavenger proteins metallothionein or MnSOD abolished the toxic effect of APP overexpression in endothelial cells. Our results demonstrate that endothelial cells are specifically susceptible to intracellular overexpression of APP and free radical generation is the likely mechanism of cell damage due to APP overexpression. PMID- 9856868 TI - Systemic administration of insulin-like growth factor decreases motor neuron cell death and promotes muscle reinnervation. AB - Neonatal sciatic nerve axotomy causes motoneuron death and muscle denervation atrophy. The aim of the present study was to determine whether insulin-like growth factor-I (IGF-I) administration promotes muscle reinnervation and counteracts motor neuron loss after such an injury. Six weeks after sciatic nerve axotomy performed in 2-day-old pups, the number of motor neurons, as assessed by retrograde transport of horseradish peroxidase injected into the extensor digitorum longus (EDL) muscle, was reduced from 52 +/- 3 to 26 +/- 3. Subsequent administration of IGF-I at the doses of 0.02 mg/kg or 1 mg/kg increased the number of motor neurons to 35 +/- 2 and 37 +/- 5, respectively. The effect on motoneuron survival was accompanied by improved muscle fibre morphometry and restoration of indirect EDL muscle isometric twitch tension, which was about 80 % of control values for both doses of IGF-I compared with 60% observed with saline treatment. Reinnervated EDL muscle from saline-treated rats cannot hold tetanic tension, which is, however, achieved after IGF-I treatment at either dose. Thus, both high and low doses of IGF-I counteracted motoneuron death and improved muscle reinnervation following neonatal sciatic nerve axotomy. IGF-I at 5 microg/kg failed to increase muscle reinnervation. PMID- 9856869 TI - Secreted phospholipase A2-induced neurotoxicity and epileptic seizures after intracerebral administration: an unexplained heterogeneity as emphasized with paradoxin and crotoxin. AB - After intracerebral injection, some toxic secreted phospholipases A2 (sPLA2) can induce epileptic seizures which bases are currently ill known. We undertook the detailed study of the central neurotoxicity of paradoxin (PDX), an analog of taipoxin, in rodents. Since literature strongly suggests a high variability in the sPLA2 epileptogenic properties, we compared, in an acute model, PDX with crotoxin (CTX), known to induce seizures and that may bind to similar neuronal receptors. Related toxic enzymes (ammodytoxin A, ATX A, and CTX subunit CB) and the non neurotoxic sPLA2 from pancreas and PLA2 analog ammodytin L (AML) were also tested. Despite being highly neurotoxic, PDX did not induce either convulsions or long-lasting seizure fits. The results obtained with the other enzymes showed that toxic sPLA2s can effectively be differentiated based on two criteria: the presence of cortically recorded epileptic paroxysmal discharges (E) and convulsions (C). We thus propose to classify the toxic sPLA2s into different groups depending on their epileptogenic properties: E-C-(PDX), E+C+ (CTX, CB), and E-C+ (ATX A). The non toxic AML and pancreatic enzyme were E-C-. Moreover, the results obtained with AML, and preliminarily with chemically inhibited CB, suggested that phospholipid hydrolysis is important to trigger seizures and convulsions. However, PDX and CTX that possess highly different epileptogenic properties exerted comparable, although slightly different, catalytic activities. Similarly, histological evaluations of the brain of PDX and CTX-treated rats (H&E staining, GFAP immunodetection, hsp70 and c-fos mRNA detection) did not provide satisfactory clues to explain these large differences. Further studies are strongly required. PMID- 9856870 TI - Antisense oligonucleotide to GABA(A) receptor gamma2 subunit induces limbic status epilepticus. AB - Gamma-Aminobutyric acid (GABA) is the principal inhibitory neurotransmitter in the brain. A deficiency of GABAergic inhibition mediated via the GABAA receptor complex has for a long time been suspected to be a central factor in epileptogenesis. Status epilepticus is a condition of sustained and prolonged excitation of neuronal circuits, as detected by epileptiform discharges in the electroencephalogram (EEG). Reduction of GABAA receptor-mediated hippocampal inhibition has been implicated in the development of status epilepticus. The present study provides direct evidence of a link between the GABAA receptor and epilepsy. We show that selective inhibition of the expression of the GABAA receptor gamma2 subunit in the rat hippocampus by means of antisense oligonucleotides leads to spontaneous electrographic seizures that evolve into profound limbic status epilepticus, ultimately resulting in severe neurodegenerative changes. Concurrent treatment with diazepam prevents the development of status epilepticus and markedly reduces neuronal cell loss. These findings strongly support the hypothesis that the GABAA receptor is critically involved in the pathogenesis of seizures and status epilepticus. PMID- 9856871 TI - Potentiated glucose deprivation-induced death of astrocytes after induction of iNOS. AB - Astrocytes play an essential role in the maintenance of normal neuronal function. Here we report that pretreatment of interferon-gamma (IFN-gamma) and lipopolysaccharides (LPS) made murine astrocytes highly vulnerable to glucose deprivation-induced death. Neither 12-hr glucose deprivation nor 2-day treatment with IFN-gamma (100 U/ml) and LPS (1 microg/ml) altered the viability of astrocytes. However, significant death of IFN-gamma/LPS-treated astrocytes was observed after 4-hr glucose deprivation. This augmented death was mimicked by the nitric oxide releasing reagent 3-morpholinosydnonimine and was in part prevented by the nitric oxide synthase inhibitor NG-nitroarginine. The data indicate that immunostimulated astrocytes can undergo suicidal death during glucose deprivation through the expression of inducible nitric oxide synthase. PMID- 9856872 TI - A new technique applicable to pediatric laparoscopic surgery: abdominal wall 'area lifting' with subcutaneous wiring. AB - BACKGROUND: Recently, the authors developed a unique method of laparoscopic surgery without pneumoperitoneum: "area lifting of the abdominal wall with subcutaneous wiring." METHODS: In this gasless procedure, the anterior abdominal wall is pulled upward by a pair of wires placed subcutaneously and held by thick sutures for "hanger lifting." Simultaneous lifting of a pair of subcutaneous wires across the abdomen, produces a wide, roof-shaped intraabdominal space sufficient for laparoscopic surgical procedures. The practical aspects of this gasless technique, as well as the authors' limited experience with this method in 24 children, ranging from 8 days to 15 years of age is presented. These children have had various pathologies including splenomegaly, rectal prolapse, ovarian cyst, gall stone, adrenal neuroblastoma, and abdominal wall abscess. CONCLUSIONS: Gasless laparoscopic surgery with double subcutaneous wiring is safe for children including neonates and those with respiratory compromise because all operative procedures are performed under normal abdominal pressure. Because of the highly elastic abdominal wall musculature inherent in children, this selective area lifting of abdominal wall creates a relatively larger peritoneal volume than in adults. PMID- 9856874 TI - The involvement of two or more systems and the severity of associated anomalies significantly influence mortality in esophageal atresia. AB - PURPOSE: The aim of this study was to examine the influence of associated anomalies in babies born with esophageal atresia (EA). METHODS: A retrospective review of the records of 41 consecutive cases of esophageal atresia managed over an 11-year period was undertaken. RESULTS: A higher incidence of associated anomalies was seen in those babies with lower birth weights. Although all five (100%) babies with EA who weighed less than 1,800 g had associated anomalies, those who weighed 1,800 to 2,500 g and more than 2,500 g were associated with 67% (10 of 15) and 43% (9 of 21) anomalies, respectively. The most common system in which anomalies occurred was the cardiovascular system (37%) followed by gastrointestinal (24%), musculoskeletal (17%), genitourinary (7%), chromosomal (5%), and others (12%). All 17 (41%) babies with no associated anomalies survived. Four of the 10 babies who had two or more systems involvement died, whereas only one of 31 babies with less than two systems involvement died; the difference between these two groups was highly significant (Fisher's Exact test, P = .009). The overall mortality rate was 12%. Three of the deaths were associated with severe anomalies that were incompatible with life such as bilateral renal agenesis, trisomy 18, and complex cardiac anomalies. CONCLUSION: The association of two or more system anomalies and the severity of associated anomalies influence mortality in esophageal atresia. PMID- 9856873 TI - Treating the snakebitten child in North America: a study of pit viper bites. AB - BACKGROUND/PURPOSE: Snakebite envenomation is a potentially life-threatening form of trauma, the dangers of which are amplified in children because their smaller size increases the relative dose of venom received. The authors reviewed a large series of snakebitten children to address the medical and fiscal issues of treating these patients. METHODS: The records of 37 snakebitten children (1987 through 1997) were analyzed for demographic data, signs of envenomation, use of specific therapies (antivenin, blood products, or surgery), length of hospitalization, complications, and cost of care. RESULTS: Fifty-four percent of the children had a major envenomation demonstrated by systemic symptomatology, laboratory analysis, or need for surgery. All children made full recoveries with most receiving only supportive care (92%). The average time to emergency department presentation was 8 hours, where all children with major envenomations and those requiring specific therapies (surgery, clotting factors) were identified. Cost analysis showed an average of $2,450 dollars per child with the majority of expenses attributable to length of hospitalization. CONCLUSIONS: Most snakebitten children completely recover with minimal supportive care, and they can be cared for safely and cost effectively as outpatients if no signs of major envenomation are noted within 8 hours of the bite. PMID- 9856875 TI - Prenatally diagnosed cystic lymphangioma in infants. AB - BACKGROUND/PURPOSE: Prenatally diagnosed cystic lymphangioma (CL) is often associated with chromosomal anomalies, hydrops fetalis, and, in the case of cervical CL, occasionally respiratory distress just after birth. Often it is difficult to treat prenatally diagnosed CL because of the large size and associated symptoms. METHODS: Between 1988 and 1997, 11 cases of prenatally diagnosed CL were treated. Five pregnancies were terminated electively and one aborted spontaneously (nondelivered, non-D group), and another five delivered (delivered, D group). Thirteen infants nondiagnosed prenatally were also seen in the same period (prenatally nondiagnosed, without pre-D group). RESULTS: In the non-D group, massive CLs were diagnosed by ultrasound scan between 16 and 20 weeks' gestation. They occurred in the head and neck in all six cases, and four had associated pleural effusions or hydrops. In the D group, CL occurred in the face and neck in four cases. Three were born by cesarean section with neonatologists and surgeons standing by, then transferred to our hospital immediately after birth. Respiratory distress appeared in two cases; tracheostomy was carried out in one case. One patient with a huge CL in the chest and abdominal wall necessitated partial resection after OK-432 sclerotherapy. In the without pre-D group, there was CL in the face or neck in 10 cases, abdominal wall in two cases, and chest wall in one case. Respiratory distress was observed in three cases of cervical CL, and tracheostomy was required in two cases. One patient with hypoxic brain damage sequela died of pneumonia afterward. CONCLUSIONS: Prenatal diagnosis permits planned delivery, prompt postnatal resuscitation, and improve prognosis. Infants with massive CL diagnosed early in pregnancy have other associated anomalies as well as hydrops fetalis, and a poor outcome or difficult management can be expected. PMID- 9856876 TI - Extracorporeal membrane oxygenation for nonneonatal pulmonary and multiple-organ failure. AB - PURPOSE: Extracorporeal membrane oxygenation (ECMO) is an accepted therapy for neonatal pulmonary failure, but its use in older children has been controversial. METHODS: Over 13 years, 55 children (ages, 3 months to 16 years) were treated with venoarterial or venovenous ECMO. The diagnoses were viral, bacterial, or fungal pneumonia (24 patients); hydrocarbon or gastric aspiration (n = 10); adult respiratory distress syndrome (ARDS), sepsis, near drowning (n = 15); pulmonary contusion (n = 2); airway obstruction (n = 3); pulmonary artery foreign body (n = 1). Pre-ECMO blood gas ranges (and means) were PO2, 21 to 100 (n = 44); PCO2, 23 to 125 (n = 72); pH, 6.81 to 7.55 (n = 7.11). RESULTS: All patients received inotropes, and 38 required dialysis or hemofiltration. ECMO was used for 20 to 613 hours (mean, 196 hours). Patient complications included cannulation site hemorrhage (n = 40), renal failure (n = 10), seizures (n = 8), stroke (n = 3), and cerebral hemorrhage (n = 2). Twenty-five patients (45%) survived ECMO, with 21 long-term survivors (10 pneumonia, five aspiration, five ARDS, one pulmonary contusion), five of whom have mild to moderate neurological deficit. Patients with combinations of pulmonary, cardiac, and renal failure, or sepsis did not survive. CONCLUSIONS: ECMO is an invasive technique that can be life saving in the child with isolated respiratory failure, but its usefulness in children with multiorgan failure is less certain. PMID- 9856877 TI - A multicenter trial of 6-aminocaproic acid (Amicar) in the prevention of bleeding in infants on ECMO. AB - BACKGROUND/PURPOSE: Intracranial hemorrhage (ICH), is a major source of morbidity and the leading cause of death in neonates treated with extracorporeal membrane oxygenation (ECMO). Anecdotal reports have suggested that epsilon-aminocaproic acid (EACA) can decrease the risk of ICH. The purpose of this study was to evaluate, in a multiinstitutional, prospective, randomized, blinded fashion, the effect of EACA on the incidence of hemorrhagic complications in neonates receiving ECMO. METHODS: All neonates (except congenital diaphragmatic hernia) who met criteria for ECMO at three institutions were eligible for enrollment. EACA (100 mg/kg) or placebo was given at the time of cannulation followed by 25 mg/kg/h for 72 hours. Bleeding complications, transfusion requirements, and thrombotic complications were recorded. Post-ECMO imaging included head ultrasound scan computed tomography (CT) scan, and duplex ultrasound scan of the inferior vena cava and renal vessels. RESULTS: Twenty-nine neonates were enrolled (EACA, 13 and placebo, 16). Five (17.2%) patients had a significant (grade 3 or larger) ICH. There was no statistical difference in the incidence of significant ICH in patients who received EACA (23%) versus placebo (12.5%). Septic patients accounted for all of the ICH in the EACA group. Thrombotic complications (aortic thrombus and SVC syndrome) developed in two patients from the placebo group. There was no difference in thrombotic circuit complications between groups. CONCLUSIONS: Our results suggest that the use of EACA in neonates receiving ECMO is safe but may not decrease the overall incidence of hemorrhagic complications. PMID- 9856878 TI - Outcome of Nissen fundoplication using intraoperative manometry in children. AB - BACKGROUND: Intraoperative manometry is useful in performing Nissen fundoplication (NF) in children. Long-term clinical outcome information after use of this method is lacking. METHODS: A retrospective review of the outcomes of 62 consecutive NFs using intraoperative manometry was performed. The follow-up period was 3.4 years. Approximately half of the patients were neurologically normal (NN) and half were neurologically impaired (NI). All patients with gastroesophageal reflux disease (GERD) did not respond to an adequate trial of medical treatment. RESULTS: The NF was tailored to result in a twofold increase in the lower esophageal sphincter pressure (LESP) and a 75% increase in the LES length (LESL). An accelerated growth rate in 40% of "failure to thrive" (FTT) patients was demonstrated. Eighty-four percent of caregivers reported improved quality of life after NF. There was a twofold reduction in the number of hospital admissions and a sixfold reduction in total inpatient days for both NI and NN children. The early and late mortality rate was 13%, and the complication rate was similar to other series reported in the literature, with more complications occurring in NI patients. There was a 2% incidence of wrap herniation. An improvement in long-term outcomes after NF was seen in 89% of NN children and over half of NI patients. CONCLUSIONS: Intraoperative manometry is useful in standardizing the tightness of the wrap in NF. There was a low incidence of complications, dysphagia, recurrent emesis, and GERD in this series. Long-term outcomes using this technique were deemed very good based on caregivers' responses. PMID- 9856879 TI - Mechanisms underlying the antireflux effect of Nissen fundoplication in children. AB - BACKGROUND/PURPOSE: It is reported that the main mechanism responsible for gastroesophageal reflux (GER) is transient lower esophageal sphincter (LES) relaxation in children. However, the effect of Nissen fundoplication on transient LES relaxation has not been investigated in children. This study examined the effect of Nissen fundoplication on motor patterns of the LES in children with pathological GER. METHODS: Esophageal manometry and pH were recorded concurrently for 2 hours after administration of apple juice (10 mL/kg). In seven children documented to have pathological GER by prolonged esophageal pH monitoring (%time pH less than 4.0>5.0), studies were performed preoperatively and 1 to 3 months after surgery. RESULTS: Nissen fundoplication virtually eliminated reflux in all patients. Percentage of time pH was less than 4.0 reduced from 15+/-9 to 0+/-0. Basal LES pressure did not change significantly (pre, 21+/-10 mm Hg v post, 27+/ 9 mm Hg). The number of transient LES relaxation reduced significantly from 13+/ 4 to 7+/-7, and the mean nadir LES pressures during swallow-induced LES relaxation and transient LES relaxation increased significantly from 1+/-1 mm Hg to 13+/-5 mm Hg and from 0+/-0 mm Hg to 11+/-7 mm Hg, respectively. CONCLUSIONS: Our findings suggest the antireflux effects of Nissen fundoplication may be based on changes of LES motor patterns that result in incomplete LES relaxation and reduction of the number of transient LES relaxation. PMID- 9856880 TI - Persistent gastroesophageal reflux disease after antireflux surgery in children: I. immediate postoperative evaluation using extended esophageal pH monitoring. AB - PURPOSE: There is a paucity of quantitative and reproducible follow-up data on childhood operations for gastroesophageal reflux disease (GERD). With the development of minimally invasive techniques for antireflux operations in children, there is a need to quantitatively determine immediate outcomes for such operations performed by laparotomy for comparison. METHODS: A retrospective review of 385 children (age range, 1 week to 15 years) who had a primary antireflux operation in a Children's or University Hospital performed by laparotomy between 1983 and 1997, and who also had an extended esophageal pH study performed within the first 12 postoperative weeks, was conducted. The operations performed included Nissen fundoplication (n = 135), Thal fundoplication (n = 195), and Boerema gastropexy (n = 55). An immediate postoperative failure of the operation to control GERD was defined as an abnormal esophageal pH score persisting up to the twelfth postoperative week. RESULTS: Eleven patients (2.9%) were classified as having an immediate postoperative failure of their operation to control GERD. An additional three patients had an abnormal esophageal pH score 2 weeks postoperatively, which subsequently reverted to a normal esophageal pH score by 12 weeks. The immediate postoperative failure rate was 1.5% (2 of 135) for the Nissen fundoplication, 1.5% (3 of 195) for the Thal fundoplication, and 10.9% (6 of 55) for the Boerema gastropexy. A higher failure rate (five patients, 36%) was seen for the first 14 patients who underwent a Boerema gastropexy during the learning curve period for this operation before 1985, and by excluding these patients the failure rate was 2.4% (1 of 41) after 1985. There was no significantly increased probability of immediate postoperative failure in patients with central nervous system disorders, prematurity, repaired esophageal atresia, or gastric emptying abnormalities. Only 5 (36%) of the 14 children with persisting symptoms suggestive of GERD had immediate postoperative failure of their operation. CONCLUSIONS: Extended esophageal pH monitoring during the first 12 postoperative weeks is a helpful tool to assess the immediate outcome of antireflux operations in children because clinical symptoms alone may be unreliable. The immediate failure rate for an antireflux operation performed in children by laparotomy is very low and seems to be unaffected by comorbid factors. PMID- 9856881 TI - Endoscopic variceal ligation in the management of gastroesophageal varices in postoperative biliary atresia. AB - BACKGROUND/PURPOSE: Gastroesophageal variceal bleeding is a serious and difficult problem in the long-term management of biliary atresia (BA). Recently, endoscopic approaches have been attempted to manage this problem. The authors have attempted endoscopic variceal ligation (EVL), a less invasive procedure than endoscopic sclerotherapy. METHODS: In the past 5 years, 66 EVL procedures using standard flexible endoscope with a diameter of 9 mm (type p-30, XQ200, or XQ240; Olympus, Tokyo, Japan) were performed in 30 separate sessions on 11 postoperative BA patients. The mean age of the children was 7.8 (range, 3 to 15) years. The EVL device was a small elastic O-ring or a loop ligator. RESULTS: EVL was performed for emergency hemostasis in two patients and prophylaxis for impending rupture in nine with large, blue varices, or with red spots on the variceal surface. During the initial procedure, all varices were ligated successfully, and reduction in size was noted. Of eight patients who were examined 7 to 14 days after treatment, seven (87.5%) had improved. Eight of 11 patients (72.7%) were finally cured or at least had improved after one to seven sessions of EVL. However, three patients did not show improvement after four to seven sessions because of the reappearance of the varices, development of distal lesions such as gastric varices, and acute gastric mucosal lesions. A technical complication encountered was a slippage of the O-ring in one patient. A technical difficulty was seen in ligating the giant gastric varix in one patient. There was no deterioration of liver function induced by EVL in this entire series. CONCLUSIONS: EVL is an effective and feasible treatment of gastroesophageal varices in postoperative BA patients. However, reappearance or reactivation of the varices or emergence of the more distal lesions is likely to occur even after repeated EVL. PMID- 9856882 TI - Jejunoileal atresia: a 27-year experience. AB - PURPOSE: In this study, the authors review cases of jejunoileal atresia (JIA) to evaluate their surgical treatment strategy. METHODS: Eighty-eight neonates who underwent surgical repair for JIA were divided into four groups for the type of lesion: group 1, membranous (n = 23), group II, interrupted (n = 49), group III, multiple (n = 9), and group IV, apple-peel (n = 7). Group I patients were treated with membranectomy or bowel resection and anastomosis, group II with resection of the dilated bowel and one anastomosis, group III with two to six multiple anastomoses to preserve bowel length, and group IV with minimal bowel resection and bowel anastomosis. During surgery a uniform protocol was used to minimize bowel resection and to perform an end-to-end single layer anastomosis using either Halsted horizontal mattress or conventional interrupted sutures. Mortality, morbidity, days for functional recovery, and central venous nutrition (CVN) were included in the review. RESULTS: Of 88 patients, three died of causes unrelated to operation for JIA. Nine patients underwent an additional laparotomy for leakage (n = 4) and obstruction (n = 5). Oral feeding was allowed on day 5.4+/-4.3 and full caloric intake via the enteric route on day 12.5+/-10.0. Twenty-one patients required CVN for 32.4+/-19.1 days. None required a long-term treatment for the short bowel syndrome. CONCLUSION: This study concludes that efforts to preserve bowel length are laudable to avoid the short bowel syndrome and that an end-to-end single layer anastomosis contributes to early recovery of bowel function. PMID- 9856883 TI - Subtotal duodenectomy with jejunal patch for megaduodenum secondary to congenital duodenal malformation. AB - BACKGROUND/PURPOSE: This report describes a technique devised for two children with megaduodenum consisting of subtotal duodenal resection with the proximal jejunum used as an onlay patch. METHODS: A 2-year-old boy presented after unsuccessful surgical repairs for duodenal stenosis with web. Another 8-year-old boy returned from the United States after several surgical procedures for complicated congenital left diaphragmatic hernia with gastric outlet obstruction. Both boys manifested postprandial right upper abdominal fullness, occasional vomiting, abdominal pain, and failure to thrive. Upper gastrointestinal contrast study findings showed massively dilated duodenums in both patients that were larger than the stomach. During the operative procedures, about 95% of the duodenal wall was resected, leaving the basal portion that was unified with the pancreatic head and the ampulla of Vater as linguiform flap, which was covered with the proximal jejunum as an onlay patch so as to form the anterior wall of the duodenal bulb. RESULTS: The patients showed markedly improved quality of life and catch-up growth after the operations. The duodenal bulbs were of adequate size in follow-up gastrointestinal series. CONCLUSION: This technique will be useful in cases of functional duodenal obstruction with megaduodenum secondary to congenital duodenal malformations leading to blind loop syndrome. PMID- 9856884 TI - The combined method: a novel access technique for fetal endoscopic surgery. AB - BACKGROUND/PURPOSE: To develop practical and less invasive techniques for fetal endoscopic surgery, new methods of lifting the uterine wall to allow fetal surgery without maternal laparotomy were developed and assessed. METHODS: Fetal endoscopic surgical procedures, including tracheostomy and umbilical vascular cannulation, were performed using one of the three methods to enter the uterus without maternal laparotomy in pregnant goats (n = 6; 105 to 115 days' gestation): (1) direct uterine lifting with an air-cushion device; (2) indirect uterine lifting, in which the uterine wall was fixed to the maternal abdominal wall using balloon tip ports inserted percutaneously by Seldinger's method, then the maternal abdomen was lifted mechanically; and (3) combined method, in which low pressure CO2 (5 mm Hg for initial inflation and 2 mm Hg for maintenance) was insufflated into the uterus in addition to the indirect uterine lifting cited above. RESULTS: The direct uterine lifting caused massive injury of myometrium and uterine membranes. The creation of intrauterine space and the protection of the membranes were not accomplished effectively by the indirect uterine lifting only. The combined method provided the adequate intrauterine space and excellent endoscopic visibility for completion of the endoscopic procedures with minimal uterine injury. CONCLUSION: The fetal endoscopic surgery may be accomplished simply and safely by the combined method, a novel technique of uterine lifting to allow fetal surgery without maternal laparotomy. PMID- 9856885 TI - Definitive localization of isolated tracheoesophageal fistula using bronchoscopy and esophagoscopy for guide wire placement. AB - PURPOSE: To aid in identification of isolated tracheoesophageal fistulas (TEF), many surgeons have recommended the bronchoscopic placement of a ureteric or Fogarty catheter. This method can fail because of intraoperative dislodgment of the catheter. The authors present a new technique that enables us to definitively isolate and treat all H-type fistulas. METHODS: Six cases of isolated TEF are presented consisting of 4 H-type fistulas, a proximal pouch fistula, and a recurrent TEF. Three of the patients had undergone a total of four prior failed operations at outside institutions using attempted bronchoscopic catheter placement. On all six patients, bronchoscopy was first performed where the fistula tract was noted in the trachea and a guide wire was passed through the fistula. After orotracheal intubation, the authors performed rigid esophagoscopy; the guide wire was identified and brought out through the mouth. This created a wire loop through the fistula. With the use of x-ray we were then able to visualize the level of the fistula and determine whether a cervical or thoracic approach should be used. Identification of the fistula intraoperatively was then facilitated by traction on the loop by the anesthesiologist. RESULTS: Five of the six TEFs were repaired with neck exploration; one required right thoracotomy. In all patients, the fistula was identified and divided. There were no recurrences or other complications. CONCLUSION: This new technique is a simple and definitive method in identification and treatment of isolated TEF. PMID- 9856886 TI - Pulmonary sequestration presenting as mitral valve insufficiency. AB - Although pulmonary sequestrations commonly present with infectious complications, problems relating to high blood flow through the lesion are rarely apparent. A 4 year-old girl was referred for cardiac catheterization and evaluation for mitral valve surgery. An echocardiogram had demonstrated left atrial and ventricular enlargement and significant mitral regurgitation with an enlarged valve annulus. Angiography results showed a very large aorta to left atrial shunt through an unsuspected intralobar sequestration. Lobectomy with removal of the sequestration resulted in significant improvement in cardiac chamber size and function over a 2.5-year follow-up period, thus obviating the need for cardiac surgery and removing a potential source of infection. Careful evaluation of chest imaging studies will lead to the correct diagnosis and treatment in patients with pulmonary sequestration who are thought initially to have primary cardiac disease. PMID- 9856887 TI - Foreign body aspiration in children: value of radiography and complications of bronchoscopy. AB - PURPOSE: The authors undertook a 10-year review of bronchoscopies for airway foreign bodies in children to test assertions in previous reports that (1) characteristic abnormalities in chest radiographs are important indicators of the need for bronchoscopy and (2) experienced operators incur negligible complications. METHODS: The medical records of 293 children who underwent bronchoscopies by experienced pediatric surgeons for suspected airway foreign bodies were reviewed for patient age, symptoms, duration of symptoms before bronchoscopy, prebronchoscopy radiographs, type of foreign body, anatomic location of foreign body, success of bronchoscopic removal, length of hospital stay, and complications. RESULTS: Of the 293 bronchoscopies, 265 showed airway foreign bodies. A choking episode was recorded in 228 of the 265 patients with foreign bodies and 5 of the 28 with negative bronchoscopy. The surgeon thought that radiographs were normal at the time of bronchoscopy in 110 patients who had foreign bodies. Nine patients with foreign bodies had atypical radiographs; three had bilateral emphysema and six had upper lobe or bilateral atelectasis or pneumonia. There were three complications: one vocal cord injury requiring temporary tracheostomy, one tracheal laceration from removal of an aspirated tooth, and one severe postoperative subglottic edema requiring a 4-day hospital stay. The age of the patients, symptoms, types of foreign bodies, locations of foreign bodies, management, and outcomes were similar to those of previous reports. CONCLUSIONS: In children with airway foreign bodies, chest radiograph findings are frequently normal and can display abnormalities uncharacteristic for foreign body aspiration. Children witnessed to choke while having small particles in their mouths and noted subsequently to have raspy respiration, wheezing, or coughing should undergo prompt bronchoscopy regardless of radiographic findings. Complications of bronchoscopy for foreign body aspiration are uncommon but occur even in experienced hands. PMID- 9856888 TI - Bronchogastric fistula, pulmonary sequestration, malrotation of the intestine, and Meckel's diverticulum--a new association. AB - Two female children, each who had a bronchogastric fistula and pulmonary sequestration (communicating bronchopulmonary foregut malformation, CBPFM) and associated malrotation of the intestine and Meckel's diverticulum are presented. Each child also presented with severe gastroesophageal reflux. The association of malrotation of the intestine and Meckel's diverticulum with a CBPFM never has been reported as a distinct entity. The concept of association of anomalies is discussed briefly. PMID- 9856889 TI - Slide tracheoplasty: a case report of successful concomitant reconstruction of extensive congenital tracheal stenosis and pulmonary artery sling. AB - An 8-month-old infant presented with an extensive congenital tracheal stenosis with an aberrant left pulmonary artery. The patient was treated successfully by relocation of the left pulmonary artery and tracheal reconstruction with slide tracheoplasty. This patient is the first survivor cited in the literature after concomitant repair using slide tracheoplasty. PMID- 9856890 TI - Preliminary evidence that cell death may contribute to separation of the trachea from the primitive foregut in the rat embryo. AB - BACKGROUND/PURPOSE: The embryogenesis of many congenital tracheoesophageal abnormalities has been elucidated poorly, mainly because of an incomplete understanding of the normal mechanism of separation of the primitive foregut into the trachea and esophagus. There has been controversy about the existence and significance of the so-called tracheoesophageal septum. This study investigates the normal mechanism of tracheoesophageal separation in the rat. METHODS: Timed pregnant Sprague-Dawley rats were killed on gestational days 11 to 16, respectively (the day on which the vaginal smear showed sperm was considered day 0 of gestation). Thirty-six embryos (six embryos from each age group) were fixed in 10% formalin. Fixed embryos from gestational day 13, 12, and 11 were sandwiched in melted agar to facilitate proper orientation before being processed. Half of the fetuses from each age group were sectioned serially in a transverse plane and the other half in a sagittal plane. The histological sections were stained with H&E. RESULTS: On gestational day 11, the foregut appears as a ventrodorsal slit in transverse section. Below the primitive pharynx, the epithelial cells on the ventral part of the foregut are actively proliferating, producing two lateral epithelial bulges (primitive bronchial buds). The epithelial cells of the dorsal aspect of the foregut show no sign of active growth. On day 12, the proximal trachea still shares a common lumen with the foregut. The distal trachea, the tracheal bifurcation, and both primary bronchi are recognized easily. At the point of tracheoesophageal separation, there is obvious debris of dead epithelial cells and condensed nuclei. These appearances were specifically located around the so-called tracheoesophageal septum and nearby grooves, resulting in enfolding cristae. There was no cell proliferation or inflammatory response in this region. On gestational day 13, the separation of the trachea and esophagus was almost complete. CONCLUSION: The present study demonstrates that a specific and consistent pattern of cell death in the region of the "tracheoesophageal septum" produced enfolding cristae that may be part of the mechanism of separation of the trachea and esophagus. PMID- 9856891 TI - Enteral glutamine does not enhance the effects of hepatocyte growth factor in short bowel syndrome. AB - PURPOSE: This study was designed to determine if luminally administered glutamine alone functions as a growth factor or is synergistic with hepatocyte growth factor (HGF) after massive small bowel resection (MSBR). METHODS: Twenty Sprague Dawley rats underwent an 80% small bowel resection and jejunostomy tube placement. Seven days later the rats were divided into four groups: group 1, control, no further treatment (n = 5); group 2 received glutamine (4% of total food intake per day) via an orogastric tube (n = 5); group 3 received intraluminal HGF via a jejunostomy tube at 75 microg/kg/d (n = 5); and group 4 received glutamine and HGF at the same doses, respectively. After a 14-day HGF infusion, glutamine feeding, or both combined, [C14] glycine absorption (micromol/L/cm2 intestine) and mucosal DNA and protein content (microg/mg mucosa) were measured in the remaining small bowel. RESULTS: Glutamine alone had no effect on substrate absorption and protein or DNA content. HGF increased galactose absorption (106% increase over control, P<.01), glycine absorption (95% increase over control, P<.05), protein content (44% increase over control, P<.01), and DNA content (32% increase over control, P<.01). The combination of glutamine and HGF did not prove to be synergistic. CONCLUSIONS: These data demonstrate that in this short bowel model, glutamine alone did not enhance intestinal function. Furthermore, glutamine is not synergistic with HGF. This study suggests that glutamine alone may not be useful clinically in patients with inadequate intestinal function. PMID- 9856892 TI - Glucose uptake in dilated small intestine. AB - BACKGROUND/PURPOSE: The development of dilated small intestine in patients with short bowel syndrome results in increased mucosal surface area. This study examines whether the incremental increase in surface area leads to a proportional increase in absorptive function of the small intestine. METHODS: Partial obstruction of the small intestine was created in rats by placing an intussusception valve in the proximal jejunum. Rats that underwent sham operations served as controls. One week postoperatively, the small intestine proximal and distal to the valve was removed. The intestinal diameter proximal and distal to the obstruction was measured. The rate of glucose uptake was measured by the everted sleeve technique. The results were analyzed by analysis of variance (ANOVA). RESULTS: The intestine proximal to the valve was significantly dilated and thickened when compared with the intestine distal to the valve. The wet mass per centimeter of the dilated segment was 2.5 times that of the control group (P<.001). The glucose uptake capacity of the dilated segment was slightly higher than that of the control group (540 v 420 nmol/min/cm, P<.05). However, the specific glucose uptake rate was reduced significantly in the intestine proximal to the valve (247 v 335 nmol/min/cm2, P<.01). CONCLUSIONS: Although the partial obstruction of small intestine resulted in a substantial increase in the intestinal surface area, the absorptive capacity of the dilated intestine per unit surface area was decreased significantly. This translated ultimately into a slight increase in the overall functional absorptive capacity of glucose in the small intestine. These results suggest that dilated small intestine may not enhance mucosal absorption. PMID- 9856893 TI - Mass screening for neuroblastoma at 6 months of age: difficult to justify. AB - BACKGROUND/PURPOSE: A statistical analysis of the mass screening for neuroblastoma in Japan based on a population study rarely has been reported. This study aims to evaluate retrospectively the effectiveness of mass screening at 6 months of age using the available population data. METHODS: The data on the neuroblastoma cases registered by the Committee for Pediatric Solid Malignant Tumors in the Kyushu area were analyzed based on both screened and unscreened populations in the Kyushu area. RESULTS: From 1988 to 1992, the cumulative incidence of neuroblastoma in children less than 5 years of age was 82 in 484,599 for screened children, and 11 in 92,966 for unscreened children, respectively. Fourteen of the 82 screened patients had negative findings at 6 months of age (MS negative cases). No significant difference was observed in the cumulative mortality rates from neuroblastoma in children younger than 5 years of age between the screened children and the unscreened children. Six of seven patients who died among the screened children were MS-negative cases with stage III or IV disease. In addition, no significant difference was found in the cumulative mortality rates from the neuroblastoma cases in patients less than 5 years of age between the children screened from 1988 to 1992 (7 of 484,599) and all children from 1980 to 1984 (14 of 668,084). CONCLUSIONS: These findings suggests that the majority of the patients detected by mass screening had a favorable prognosis, and, mass screening in Japan for children less than 6 months of age was not observed to reduce the incidence and mortality from neuroblastoma. Therefore, mass screening at 6 months of age was not found to improve substantially the prognosis of patients with unfavorable neuroblastoma identified over 1 year of age, which is the primary purpose of such mass screening for neuroblastoma. PMID- 9856894 TI - Extra-anal mucosectomy: laparascopic-assisted endorectal pull-through using a prolapsing technique. AB - BACKGROUND/PURPOSE: The mucosectomy of the aganglionic rectal mucosa has been performed transabdominally in the Soave procedure. Recently, mucosectomy with transanal approach was reported both in the laparoscopic and the open surgical procedure. However, the operative view and working field are restricted because the dissection has to be done in the anal canal. The authors report an innovative approach, an extra-anal mucosectomy, to perform mucosectomy in the Soave procedure for Hirschspung's disease in combination with the laparoscopic-assisted prolapsing technique. METHODS: Four patients underwent laparoscopic surgery for Hirschsprung's disease with extra-anal mucosectomy between 1995 and 1997. One 10 mm and 5-mm ports were used throughout the operation. The rectum was pulled out through the anal canal to create a rectal prolapse and was divided outside the anal canal. The mucosectomy was performed in the everted rectal mucosa outside the anal canal to the level of dentate line. The colon was sutured to the anal mucosa 2 mm above the dentate line. Mean operative time was 3 hours. RESULTS: The present technique made the whole mucosectomy possible under direct observation. CONCLUSION: The extra-anal mucosectomy in conjunction with a laparoscopic assisted prolapsing technique seems to be a safe and reliable modality in the surgical treatment of Hirschsprung's disease. PMID- 9856896 TI - Virtual intraluminal endoscopy: a new method for evaluation and management of choledochal cyst. AB - A 12-year-old girl who suffered from cholangitis was treated successfully with appropriate antibiotics. She had undergone an operation in mainland China, the exact nature of which was unknown. After an ultrasound study, she underwent a helical computed tomography (CT) cholangiogram using intravenous meglumine iodoxamine. The volume data were transferred to a workstation and virtual endoscopy rendering reconstruction of bile ducts was performed. Findings of this study showed that she had cyst excision with hepaticojejunostomy (HJ), and her intrahepatic ducts were packed with stones. Virtual endoscopy gave the impression that the examiner was flying inside the lumen and showed the stricture at the HJ anastomosis, the inner surface of the bile ducts, areas of dilated intrahepatic ducts, and the intrahepatic stones. The study overcame the need for an invasive study such as a percutaneous transhepatic cholangiography (PTC) and also facilitated appropriate surgical treatment in a timely fashion. It is believed that virtual intraluminal endoscopy (VIE) is helpful in the evaluation and management of selected cases of choledochal cyst. PMID- 9856895 TI - Necrotizing enterocolitis and C-KIT. AB - BACKGROUND/PURPOSE: In the gut, C-KIT is important for immune system homeostasis, and C-KIT+ cells are known to increase during inflammation. Recently the authors identified that spontaneous intestinal mucosal erosion develops in C-KIT-depleted W/Wv mice after day 14 of life at a high frequency, whereas genotypically normal litter mates do not. The authors hypothesized that a lack of C-KIT may be implicated in the development of necrotizing enterocolitis (NEC). METHODS: Bowel specimens were taken during surgery or postmortem from nine cases of NEC (mean gestational age, 32.0 weeks), six age-matched cases of enteritis, and 10 age matched controls. Specimens were formalin fixed, paraffin embedded, and labeled with antibody to C-KIT. The number of C-KIT+ cells from five random fields per specimen were compared under light microscopy (200x). Results were expressed as the mean +/- SD and compared using the analysis of variance (ANOVA) test. RESULTS: In enteritis, the number of C-KIT+ cells in the lamina propria and submucosa was significantly higher than in controls (P<.01) indicative of their involvement in inflammation. However, in NEC, the number of C-KIT+ cells in the lamina propria and submucosa was significantly lower than in controls (P<.05) despite histological evidence of inflammation. CONCLUSION: A lack of C-KIT+ cells may exert a causal influence on the development of NEC. PMID- 9856897 TI - Complete resection is not required in patients with neuroblastoma under 1 year of age. AB - BACKGROUND/PURPOSE: The prognosis of neuroblastoma (NB) patients when detected before 1 year of age is excellent, particularly in patients whose disease is detected by screening. In some institutions patients with stage I or II NB detected by screening have been observed closely without any treatment. Most showed tumor regression or maturation and are surviving without tumor resection. Resection of infantile NB sometimes is complicated by vascular accidents resulting in vanishing kidney or intestinal infarction. The role of surgery in the treatment of infantile NB therefore is becoming controversial. The authors have treated infants who have easily resectable primary tumors with resection and sampling for lymph node metastases. If the resection of the primary tumor seemed difficult, only biopsy was performed initially followed by chemotherapy with or without resection. The authors analyzed the clinical outcome of the infantile NB in their institution to elucidate the role of surgery in this particular group of patients. METHODS: Thirty-six patients less than 12 months of age were treated between 1982 and 1997. Twenty-five patients who showed no symptoms had NB detected by screening at 6 months. Five patients had stage IVS disease in early infancy. No patients had bone or remote lymph node metastases. N-mycamplification was not detected in any of the 28 tumors examined. There were four diploid and 12 near-triploid tumors. RESULTS: Complete resection was possible in 13 stage I and II patients without any complications. Obvious lymph node metastases were not resected in 13 patients with stage II or III NB. Partial resection leaving gross residual disease was accomplished in three patients with stage III NB. Surgical resection was not attempted in four patients because of unresectability in three patients with stage III or IVS disease and because of marked tumor shrinkage after a short course of chemotherapy in one patient with stage III NB involving the celiac axis. A neonatal cervicomediastinal stage III NB recurred very rapidly after partial resection but started to regress spontaneously 2 months postoperatively. Another neonatal stage II cervical NB progressed to stage IVS with hepatomegaly causing respiratory distress but regressed spontaneously. All the patients are alive and disease free without surgical morbidity. CONCLUSION: All of the 21 patients who underwent partial tumor resection with or without chemotherapy are alive and doing well without tumor, indicating that complete resection is not required in this particular group of patients. PMID- 9856898 TI - Fluorescence in situ hybridization analysis of chromosome 1p36 deletions in human MYCN amplified neuroblastoma. AB - BACKGROUND/PURPOSE: Deletion of the short arm of chromosome 1 (1p) is one of the poor prognostic factors in human neuroblastomas. Recent studies have suggested that one or more of the neuroblastoma tumor suppressor genes reside in this region and have identified the shortest region of overlap (SRO) on 1p36. The purpose of this study was to examine deletions of 1p in human neuroblastomas by fluorescence in situ hybridization (FISH). METHODS: Two-color FISH analysis was performed to detect chromosome 1p36 abnormalities in 42 MYCN-amplified neuroblastomas. Four different probes from the 1p36 region, the E2F2, NPPA, D1S160, and CDC2L1 loci were used for detection of 1p abnormalities. A repeat sequence probe, which is specific for the heterochromatic region of chromosome 1 (pUC1.77), was used as a control. RESULTS: Large deletions of 1p36 were observed in 31 (73.8%) of 42 tumors, whereas the remaining 11 (26.2%) showed no deletion. In these 11 tumors, a translocation of 1p was found in one and a duplication of 1p was detected in another. CONCLUSIONS: A strong correlation between 1p abnormalities and MYCN amplification was found in this study. MYCN-amplified neuroblastomas were found to show large deletions of 1p encompassing the SRO. FISH provided a rapid and reliable method to detect hemizygous deletions of 1p. PMID- 9856899 TI - Efficacy of exogenous fibronectin in wound healing in malnourished rats. AB - BACKGROUND/PURPOSE: Fibronectin (FN) plays an important role in fibrin matrix formation during the wound healing process. The authors investigated whether exogenous FN increases the bursting strength (BS) of surgical wounds in malnourished rats. METHODS: Ninety rats were grouped according to three nutritional conditions (n = 30 in each group). All animals underwent a transverse celiotomy after 3 weeks of feeding, and FN (2 mg/body/day) was given postoperatively to 15 rats in each of the three groups. Wound BS was measured on postoperative days (PODs) 3, 5, and 7 after removal of the sutures. RESULTS: BS in normally nourished rats (group N; 142.6+/-23.4 mmHg) was significantly higher than that in protein malnourished rats (group PM, 110.2+/-11.2 mm Hg) and protein/ calorie (Cal)-malnourished rats (group PCM, 76.5+/-10.7 mm Hg) on POD 7 (P<.01). However, BS values for groups PM + FN (147.0+/-21.1 mmHg) and PCM + FN (115.1+/-28.9 mm Hg) were intensified significantly in comparison with groups PM and PCM (P<.01). Plasma FN levels in rats of the three FN nontreated groups were similarly decreased on POD 3 or 5, but returned to the preoperative level on POD 7, whereas those for the other three FN-treated groups increased after POD 3. CONCLUSION: Intravenous administration of FN might strengthen the weakened wounds of malnourished animals. PMID- 9856900 TI - Segmental small bowel allograft--ischemic injury and regeneration. AB - PURPOSE: Small bowel transplantation (SBT) is the ultimate treatment for intestinal failure. It remains unclear as to which intestinal segment is more suitable for use in segmental SBT. The current study aims to assess the susceptibility of various parts of small intestine to ischemia and reperfusion injury and their capacity for regeneration. METHODS: Thirty-two segments of pig jejunum and ileum were isolated with intact vascular pedicles that were clamped for periods varying from 1/2 to 8 hours. Biopsy specimens were taken immediately before clamp release and 20 minutes afterwards. All segments were anastomosed together before abdominal closure. Laparotomy was performed 24 hours later, and biopsy specimens were taken at all segments. All specimens were examined histologically by a pathologist. RESULTS: Evidence of injury was detected after 1.5 hours of ischemia at jejunum, but only after 5 hours at ileum. More severe injury was noted at the initial period on reperfusion, but there was no further deterioration at the later period. Complete reepithelialization occurred after 24 hours even where there had been total villous sloughing at reperfusion, but regeneration was impossible when the crypts had been damaged completely. CONCLUSIONS: Ileum, because it is more resistant to ischemia and reperfusion injury, may be preferred for segmental SBT. Regeneration of the bowel epithelium is fast, provided that the crypts are not damaged completely. PMID- 9856901 TI - Tracheobronchial injuries after blunt chest trauma in children--hidden pathology. AB - BACKGROUND: Blunt thoracic injuries in children are unique because the pliability of the chest wall allows transmission of massive external force directly into the mediastinum. Children presenting after blunt chest trauma may have complete disruption of the airway with little external sign of injury. Without prompt diagnosis and appropriate treatment, the risk for progressive respiratory failure is high. METHODS: Four children with tracheobronchial injuries were referred to a pediatric trauma center from 1994 to 1997. All children, age 18 months to 13 years, suffered unusual crush injuries. All diagnoses were based on unresolved pneumothorax or pneumomediastinum. RESULTS: Bronchoscopy identified the location of injury as posterior trachea (n = 1) and right mainstem bronchus (n = 2). A tertiary bronchial injury (n = 1) was missed by initial tracheogram and subsequent bronchoscopy but identified during surgical exploration. All children survived after thoracotomy and primary repair of the injury. CONCLUSIONS: Tracheobronchial disruption is a rare, life-threatening injury. Suspicion should be high when pneumomediastinum and pneumothorax are refractory to adequate pleural drainage. Flexible bronchoscopy with intubation distal to the injury may be necessary to prevent loss of the airway. Advance preparation should include setups for bronchoscopy, thoracotomy, and cardiopulmonary bypass. Patient survival depends on preparation and prompt surgical intervention. PMID- 9856902 TI - Driveway crush injuries in young children: a highly lethal, devastating, and potentially preventable event. AB - BACKGROUND/PURPOSE: The aim of this study was to investigate driveway-related injuries in children, identify associated risk factors, and evaluate outcome compared with other mechanisms of blunt trauma. METHODS: A 6-year review (1991 to 1996) of pediatric (age less than 18 years) pedestrian injuries treated at two urban trauma centers was conducted: one regional pediatric trauma center and one level I trauma center with pediatric commitment. Five hundred twenty-seven children injured in pedestrian accidents were identified from the trauma registry; 51 children (10%) sustained traumatic injuries as a result of being struck in their driveway. Data are reported as mean +/- SEM. RESULTS: Children less than 5 years of age (n = 41) had an injury severity score (ISS) of 12.3+/ 2.3, 15 (37%) sustained closed head injury, 13 (37%) had torso trauma, 19 (46%) skeletal trauma, and eight (20%) died. Children > or = 5 years old (n = 10) had an ISS of 10.7+/-2.4, three (30%) sustained closed head injury, four (40%) torso trauma, six (60%) skeletal trauma, and none died. In contrast, all other pediatric pedestrian accidents analyzed over the same time period had a mortality rate of only 2% (11 of 476). CONCLUSIONS: Pediatric driveway trauma carries a significant risk of head injury and a 10-fold increase in mortality in children under 5 years of age when compared with all other pediatric pedestrian accidents. More emphasis must be placed on injury prevention and public education to prevent this devastating mechanism of injury in these young, vulnerable children. PMID- 9856903 TI - Surgical treatment of vesicoureteral reflux in infants under 3 months of age. AB - PURPOSE: Surgical treatment for vesicoureteral reflux (VUR) is controversial in infants, especially small infants because of technical difficulty and higher rate of spontaneous resolution. However, in some conditions, such as high-grade reflux, breakthrough infection, or severe renal scarring, early operation may be justified. This study is to evaluate the results of ureteral reimplantation in infants with VUR under 3 months of age. METHODS: From January 1993 to July 1997, 13 boys and five girls (24 ureters) under 3 months of age (range, 18 days to 3 months; mean age, 2 months) received ureteral reimplantation for VUR. The initial symptoms were urinary tract infection (UTI) in 16 infants, hydronephrosis found by prenatal ultrasound scan in one, and early postnatal screening ultrasound scan in one. The indications for 24 ureter reimplantations were high-grade reflux (grade V, n = 7), breakthrough infection under medical treatment (n = 5), and severe renal scarring (n = 8) and contralateral low-grade refluxing ureters (n = 4). All patients had Glenn-Anderson's or Cohen's ureteral reimplantation. Ureteral stents were required in six ureters. All patients had renal sonogram 1 month after operation to rule out ureteral obstruction. Fifteen patients had voiding cystourethrogram (VCUG) after operation. Renal growth was evaluated in 15 patients (20 reflux renal units) using serial ultrasound or differential perfusion renal scan. The operative time, use of ureteral stent, complication rate, hospital stay, and time needed for clearing urine, were compared with those of 115 counterpart patients at age over 3 months. Data were analyzed using X(2) test or t test. RESULTS: All patients, except one, were free of UTI during the follow-up of 6 months to 5 years. In fifteen infants who had postoperative VCUG, 14 were free of reflux, and one had persistent lower-grade VUR. One patient had transient ureteral obstruction that resolved spontaneously. One patient had postoperative ileus for 2 weeks. Renal growth of 20 reflux renal units was similar to that of the contralateral nonreflux kidneys. The only significant difference when compared with the patients over 3 months of age, was the higher incidence of ureteral stenting (25% v 4.2%, P<.01). CONCLUSIONS: Ureteral reimplantation should be performed in infants under 3 months of age when it is indicated. PMID- 9856904 TI - Gastric necrosis after fundoplication: a novel approach for esophageal preservation. AB - An 11-year-old boy presented moribund, with massive abdominal distension. A Nissen fundoplication and gastrostomy tube had been established at age 2 years. After attempts to pass a nasogastric tube were unsuccessful, the old gastrostomy site was used to gain percutaneous access to the stomach resulting in release of gastric contents and stabilization of blood pressure and perfusion. During operation, massive gastric distention with gastric necrosis was found. Subtotal gastrectomy was performed with stapled closure of the distal intraabdominal esophagus and prepyloric region. Sump suction was placed in the proximal esophagus and the abdomen was drained widely. A distal esophageal perforation was apparent on postoperative day 19 confirmed by imaging and endoscopy. A nasoesophageal tube was passed into the abdomen, tied to a Jackson-Pratt drain, and the composite tube repositioned in the midesophagus allowing controlled proximal and distal drainage. Six months later, a Hunt-Laurence esophagojejunal pouch was created. At age 13, the child is clinically well, and enjoys 50% of his nutritional needs orally, with the remainder delivered overnight via tube feedings. This case describes gastric necrosis after gas bloat syndrome as a late complication of Nissen fundoplication. A novel approach to the management of distal esophageal perforation allowed preservation of a functional, intact native esophagus. PMID- 9856905 TI - High oxygen delivery and extraction by perfluorocarbon-primed extracorporeal membrane oxygenation do not prevent anaerobic metabolism in rabbits. PMID- 9856906 TI - Laparoscopic excision of pelvic kidney with single vaginal ectopic ureter. PMID- 9856907 TI - Effects of anal invasive treatment and incontinence on mental health and psychosocial functioning of adolescents with Hirshsprung's disease and low anorectal anomalies. PMID- 9856908 TI - Type IV laryngotracheoesophageal clefts: surgical management for long-term survival. PMID- 9856909 TI - Reduction of prosthetic silo: a novel technique using a desktop stapler. PMID- 9856910 TI - A fourth pharyngeal pouch remnant located in the right side of the neck. PMID- 9856911 TI - Age-related trends in short- and long-term survival after acute myocardial infarction: a 20-year population-based perspective (1975-1995). AB - This study examines age-related differences and temporal trends in hospital and long-term survival after acute myocardial infarction (AMI) over a 2-decade-long (1975 to 1995) experience. A total of 8,070 patients with validated AMI hospitalized in all acute care hospitals in the Worcester, Massachusetts, metropolitan area (1990 census population 437,000) were studied over 10 one-year periods between 1975 and 1995. This population included 1,326 patients aged <55 years (16.4%), 1,768 patients aged 55 to 64 years (21.9%), 2,325 patients aged 65 to 74 years (28.8%), 1,880 patients aged 75 to 84 years (23.3%), and 771 patients aged > or = 85 years (9.6%). Compared with patients <55 years, patients 55 to 64 years were 2.2 times more likely to die during hospitalization for AMI, whereas patients 65 to 74, 75 to 84, and > or = 85 years were at 4.2, 7.8, and 10.2 times greater risk of dying, respectively. Similar age disparities in the risk of dying were seen when controlling for additional prognostic factors. Despite the adverse impact of increasing age on hospital survival after AMI, declining in-hospital death rates were seen in each of the age groups under study, with declining magnitude of these trends with advancing age. Among discharged hospital patients, increasing age was related to a significantly poorer long-term prognosis. Trends toward improving long-term prognosis were seen in patients discharged in the mid 1990s compared with those discharged in the mid- to late 1970s for patients aged <85 years. The present results demonstrate the marked impact of advancing age on survival after AMI. Despite the adverse impact of age on prognosis, encouraging trends in prognosis were observed in all age groups, although to a lesser extent in the oldest elderly patients. These findings emphasize the low death rates in middle-aged patients with AMI and the need for targeted secondary prevention efforts in elderly patients with AMI. PMID- 9856912 TI - New electrocardiographic criteria for predicting the site of coronary artery occlusion in inferior wall acute myocardial infarction. AB - In patients with inferior wall acute myocardial infarction (AMI), the site of the culprit lesion is an important determinant of outcome. Patients with right ventricular infarction have a poor prognosis, whereas those with occlusion of the left circumflex coronary artery (LCx) have a good prognosis. Therefore, we assessed whether standard 12-lead electrocardiograms obtained on admission could identify the site of coronary artery occlusion, (i.e., a site proximal to the origin of the right ventricular branch of the right coronary artery [RCA], a site distal to the origin of the right ventricular branch of the RCA, or a site in the LCx). The ratio of ST depression in lead V3 to ST elevation in lead III (V3/III ratio) was evaluated immediately before coronary angiography in 152 patients with a first inferior wall AMI confirmed by coronary angiography within 12 hours after the onset of symptoms. For occlusion of the proximal RCA, distal RCA, and LCx, V3/III ratio was 0.2+/-0.3, 0.8+/-0.5, and 2.5+/-2.5 (p = 0.0001), respectively. The V3/III ratio <0.5 identified proximal RCA occlusion, 0.5 or = 10/hour were the most powerful independent predictors for all end points, whereas no HRV was independently associated with the events. A low frequency/high frequency ratio < 1.05 only had a borderline association with sudden death (RR = 2.86, p = 0.076). Our data show a strong association between HRV and mortality in patients surviving a recent MI, with a slight better sensitivity of frequency domain analysis. In our study, however, HRV did not add independent prognostic information to more classic prognostic variables (e.g., left ventricular function and ventricular arrhythmias). PMID- 9856914 TI - Comparison of reported symptoms of acute myocardial infarction in Mexican Americans versus non-Hispanic whites (the Corpus Christi Heart Project). AB - This study examines whether there are differences between Mexican Americans and non-Hispanic whites in reported symptoms of acute myocardial infarction (AMI). The symptoms experienced by patients identified in a community-based surveillance program were examined to determine whether between-group differences existed by ethnicity, gender, and diabetic status. Data were available regarding the symptoms of 589 patients, between the ages of 25 and 74 years, who were hospitalized and diagnosed as either having definite or possible AMI in special care units at 1 of 7 hospitals in Corpus Christi, Texas. The most frequently reported symptoms were chest pain (83.2%), chest pressure or discomfort (67.6%), sweating (64.2%), fatigue (62.6%), dyspnea (60.3%), and arm or jaw pain (58.2%). After adjusting for age, diabetes mellitus, and gender, and relative to non Hispanic whites, Mexican Americans were more likely to report chest pain, upper back pain, and palpitations, and less likely to report arm or jaw pain. Likewise, relative to men, women were more likely to report fatigue, dyspnea, dizziness, upper back pain, palpitations, and cough, and were less likely to report chest pain. Significant differences were also observed when older patients' symptoms were compared with younger patients' symptoms. PMID- 9856915 TI - Prognosis of "clandestine" myocardial ischemia, silent myocardial ischemia, and angina pectoris in medically treated patients. AB - The aim of this study was to assess the prognosis of medically treated patients with "clandestine" myocardial ischemia (perfusion defect without angina and no ST depression > 1 mm during exercise test) compared to those with silent myocardial ischemia (ST-segment depression > 1 mm, without angina) and those with angina pectoris. One hundred twelve patients without previous myocardial infarction were included. All patients underwent a symptom-limited exercise test on a bicycle ergometer, myocardial perfusion technetium-99m-methoxy-isobutyl-isonitrile single photon emission computed tomography (SPECT), and coronary angiography. They were classified into 3 groups (angina group, 34 patients; silent group, 20 patients; and the clandestine group, 58 patients). The mean follow-up was 3.6 years (range 6 months to 5.5 years). Patients with clandestine ischemia had a lower scintigraphic and angiographic score than patients with silent ischemia (25+/-8 vs 31+/-9 and 24+/-8 vs 29+/-7, p = 0.008, respectively), but the prognosis was similar. Only angina and severe reversible SPECT defects were predictive for cardiac events: death + myocardial infarction + revascularization. We conclude that in medically treated patients without previous myocardial infarction, angina and severe reversible SPECT defects are predictive for cardiac events only when the need for revascularization is included as a cardiac event. PMID- 9856916 TI - Optimal criteria for the diagnosis of coronary artery disease by dobutamine stress echocardiography. AB - The diagnosis of coronary artery disease (CAD) on the basis of inducible ischemia in > or = 2, rather than 1, segments was suggested to improve specificity of dobutamine stress echocardiography (DSE). However, the impact of using these criteria on the sensitivity and accuracy of DSE was not studied. We studied the accuracy of DSE (up to 40 microg/kg/min) for the diagnosis of CAD in 290 patients with suspected myocardial ischemia using the criteria of > or = 1 and > or = 2 ischemic segments. Ischemia was defined as new or worsening wall motion abnormalities using a 16-segment model. Among the 85 patients without previous myocardial infarction, significant CAD was detected in 51 (60%). Sensitivity, specificity, and accuracy of DSE using > or = 1 ischemic segment were 73%, 85%, and 78%, respectively. Those using > or = 2 segments were 67%, 94%, and 78%, respectively (p = NS). Regional specificity improved by using > or = 2 segments (91% vs 96%, p <0.05) at the expense of an equivalent reduction in regional sensitivity (60% vs 44%, p <0.05), whereas the regional accuracy was similar (80% vs 79% for > or = 1 and > or = 2 segments, respectively). In the 205 patients with previous myocardial infarction, the criterion of ischemia in > or = 1 segment had a higher sensitivity and accuracy for overall diagnosis of CAD (75% vs 64%, p <0.05; 77% vs 68%, p <0.05), infarct-related CAD (64% vs 47%, p <0.005; 70% vs 57%, p <0.01), and remote CAD (74% vs 57%, p <0.005; 78% vs 69%, p <0.05) than the criterion of > or = 2 segments, respectively. It is concluded that in patients without previous myocardial infarction, the use of > or = 2 ischemic segments by DSE for the diagnosis of CAD does not improve the accuracy of DSE compared with the criterion of > or = 1 ischemic segment. Conversely, in patients with previous infarction the use of > or = 2 segments reduces the overall and regional sensitivity and accuracy without a significant improvement in specificity. PMID- 9856917 TI - Directional coronary atherectomy for the treatment of Palmaz-Schatz in-stent restenosis. AB - Management of in-stent restenosis has become a significant challenge in interventional cardiology. The results of balloon angioplasty have been disappointing due to the high recurrence of restenosis at follow-up. Debulking of the restenotic tissue within the stents using directional coronary atherectomy (DCA) may offer a therapeutic advantage. We report the immediate clinical and angiographic outcomes and long-term clinical follow-up results of 45 patients (46 lesions), mean age 63+/-12 years, 73% men, with a mean reference diameter of 2.9+/-0.6 mm, treated with DCA for symptomatic Palmaz-Schatz in-stent restenosis. DCA was performed successfully in all 46 lesions and resulted in a postprocedural minimal luminal diameter of 2.7+/-0.7 mm and a residual diameter stenosis of 17+/ 10%. There were no in-hospital deaths, Q-wave myocardial infarctions, or emergency coronary artery bypass surgeries. Four patients (9%) suffered a non-Q wave myocardial infarction. Target lesion revascularization was 28.3% at a mean follow-up of 10+/-4.6 months. Kaplan-Meier event-free survival (freedom from death, myocardial infarction, and repeat target lesion revascularization) was 71.2% and 64.7% at 6 and 12 months after DCA, respectively. Thus, DCA is safe and efficacious for the treatment of Palmaz-Schatz in-stent restenosis. It results in a large postprocedural minimal luminal diameter and a low rate of both target lesion revascularization and combined major clinical events at follow-up. PMID- 9856918 TI - Effects of doxazosin on exercise-induced angina pectoris, ST-segment depression, and insulin sensitivity in patients with syndrome X. AB - A significant proportion of patients with cardiac syndrome X have impaired coronary vasodilator capacity, which is thought to be caused by an increased sympathetic drive. The alpha1-adrenoceptor blocker, doxazosin, increases the coronary vasodilator reserve in patients with syndrome X. To study whether the augmentation is associated with clinical improvement in patients, we conducted a double-blind, placebo controlled, crossover study with doxazosin 1 to 4 mg once daily for 10 weeks in 16 patients with syndrome X (14 women and 2 men; mean +/- SD age 56+/-5 years). Time to angina, exercise duration, time to 0.1 mV ST segment depression, and maximal ST-segment depression during bicycle exercise testing were compared after treatment with doxazosin 2 mg or placebo for 5 weeks and again after treatment with doxazosin 4 mg or placebo for 10 weeks. Insulin sensitivity was assessed by the minimal model after 10 weeks of doxazosin or placebo treatment. Twelve patients completed the protocol. Doxazosin 4 mg/day decreased systolic blood pressure at rest (109+/-16 vs 125+/-18 mm Hg, p <0.05) and increased basal heart rate (85+/-9 vs 76+/-11 beats/min, p <0.05), whereas hemodynamics were unaffected during exercise. Time to angina, exercise duration, time to 0.1 mV ST-segment depression, and maximal ST-segment depression were similar during treatment with doxazosin and placebo irrespective of the doxazosin dose. Insulin sensitivity was not different with doxazosin and placebo. In conclusion, alpha1 blockade does not significantly improve exercise duration, angina pectoris, and ST-segment depression despite a favorable vasodilator effect in patients with syndrome X. The absent clinical efficacy of doxazosin may challenge the use of the coronary vasodilator capacity as an appropriate method to subclassify patients with syndrome X. PMID- 9856919 TI - Cost-effectiveness of statins. AB - Currently, 6 hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are marketed in the United States (US). Given the wide variation in the prices and efficacy of statins, formal cost-effectiveness analysis may improve drug selection decisions. To assess the cost-effectiveness of statin therapy in primary and secondary prevention of coronary heart disease, we developed a model of the costs and consequences of lipid-regulating therapy and estimated the incremental cost-effectiveness of 5 statins (atorvastatin, fluvastatin, lovastatin, pravastatin, simvastatin) at usual starting doses versus no therapy. Drug effects on serum lipids were assessed using data approved by the US Food and Drug Administration for product labeling. Annual risks of coronary event occurrence were estimated using Framingham Heart Study coronary risk equations developed for use in this model. Current estimates of direct medical costs of coronary heart disease were used to assign costs to health states and acute coronary events. Main outcome measurements were net cost (statin therapy minus savings in coronary heart disease treatment), gain in life expectancy, and cost per life-year saved. The maximum gain in life expectancy was achieved with atorvastatin, which also had a lower net cost than lovastatin, pravastatin, and simvastatin. Compared with fluvastatin, atorvastatin's greater effectiveness is attained at a lower cost per life-year saved. The cost-effectiveness of HMG-CoA reductase inhibition in primary and secondary prevention of coronary heart disease has been improved with the introduction of atorvastatin. PMID- 9856920 TI - Relation between aging and circadian variation of paroxysmal atrial fibrillation. AB - The purpose of this study was to determine whether aging influences the circadian variation of nonvalvular paroxysmal atrial fibrillation (AF). Among 31,200 consecutive Holter monitorings recorded between January 1988 and March 1997, we detected 212 patients who had paroxysmal AF in a drug-free state. These patients were divided into 2 groups according to their age: < or = 60 years old (94 patients) and >60 years old (118 patients). In each group, the sum of the duration of each AF episode and the probability of onset, maintenance, and termination of AF were determined as hourly data and compared between the 2 groups. The time distribution of AF showed remarkable age dependence, with a well modulated and monophasic circadian rhythm in the younger group in contrast to a toneless triphasic rhythm in the older group. Among the onset, maintenance, and termination of the arrhythmia, the most obvious age-dependence was observed in the circadian variation of onset. In the younger group, there were triple peaks with the highest one in the night, whereas the older group exhibited a single peak in the daytime. In contrast, the probabilities of maintenance and termination showed similar circadian patterns between the groups, although their amplitudes were significantly reduced in the older group. Thus, aging significantly influenced the circadian variation of paroxysmal AF, with the most prominent effect on its onset, leading to more random time-distribution of AF with increasing age. These results extend to paroxysmal AF the concept that aging disrupts rhythmicity, suggesting age-dependent differences in its pathophysiology. PMID- 9856921 TI - Clinical and echocardiographic features influencing recovery of atrial function after cardioversion of atrial fibrillation. AB - Atrial mechanical dysfunction after cardioversion for atrial fibrillation has been widely evaluated in recent years. Nevertheless, the influence of many clinical and echocardiographic parameters is not yet understood. The aim of the present study was to evaluate the influence of clinical and echocardiographic parameters on the return of effective atrial contraction. A total of 109 patients were evaluated: 41 patients had spontaneous recovery of sinus rhythm and 68 patients were randomly treated using either direct-current (DC) shock or intravenous procainamide. Elective cardioversion was accomplished pharmacologically in 23 patients (67%) and with DC shock in 29 patients (85%). Patients underwent a complete echocardiographic examination 1 hour after the restoration of sinus rhythm and after 1 and 7 days and 1 month. The following parameters were evaluated: patient age, cardiac disease, duration and etiology of atrial fibrillation, mode of cardioversion, left ventricular diameters and function, and left atrial diameter and function assessed as atrial ejection force. The relation between these variables and atrial ejection force was tested. Atrial ejection force was greater immediately and 24 hours after cardioversion in patients who had spontaneous recovery of sinus rhythm and in patients treated with drugs than in patients treated with DC shock. The mode of cardioversion was significantly associated with the recovery of atrial mechanical function by day 1 in univariate and multivariate analyses (odds ratio 0.14; 95% confidence interval 0.02 to 1.2). The other variable associated with the recovery of function was normal left atrial size (odds ratio 0.16; 95% confidence interval 0.12 to 1.6). In conclusion, atrial ejection force is a noninvasive parameter that can be easily measured and can provide accurate information about the recovery of left atrial mechanical function. The recovery of atrial function was influenced by the mode of cardioversion and the size of the left atrium. PMID- 9856922 TI - Mechanisms of syncope in implantable cardioverter-defibrillator recipients who receive device therapies. AB - A significant proportion of implantable cardioverter defibrillator (ICD) recipients ultimately receive device therapies. Determinants and mechanisms of syncopal events in these patients are not established. To address this issue, we reviewed prospectively collected data on 114 ICD recipients who received device therapies. There were 99 men and 15 women with a mean age of 63+/-10 years. Ejection fraction was 29+/-13%. At initial presentation, 74 patients (65%) had syncope during arrhythmia. Of 114 patients, 18 patients (16%) subsequently experienced syncope during device therapies over a period of 35+/-24 months of follow-up. Arrhythmias during syncopal events documented by telemetry, event recorders, RR intervals, or ICD stored electrograms were obtainable in 13 patients, and included sustained monomorphic ventricular tachycardia (VT) in 10 patients, ventricular fibrillation (VF) in 2 patients, and atrial fibrillation in 1 patient. The explanations for arrhythmia-related syncope included ICD proarrhythmia in 8 patients (62%), rapid monomorphic VT in 3 (23%), and VF in 2 patients (15%). Patients with syncope could not be differentiated from those without syncope by demographic, clinical, or electrophysiologic variables. There was no significant difference in mortality between patients with or without syncope (39% vs 25%, p = 0.25). Syncope is not uncommon in ICD recipients who receive device therapies. When it occurs, syncope is often due to ICD proarrhythmia; hence, it is potentially preventable by meticulous device programming. PMID- 9856923 TI - Pattern of changes over time in myocardial blood flow and microvascular dilator capacity in patients with normally functioning cardiac allografts. AB - This study tests the hypothesis that myocardial blood flow and coronary microvascular dilator capacity vary as a function of time after orthotopic heart transplantation in humans. Positron emission tomography measurements of myocardial blood flow were obtained at rest and during adenosine in 24 patients between 1 and 86 months after heart transplantation. At the time of the study all patients were clinically well and had angiographically normal epicardial coronary artery vessels. Patients were divided into 3 groups based on time from transplant to positron emission tomography measurement of myocardial blood flow: group 1 to 12 months (n = 9); group 13 to 34 months (n = 8); and group > or = 37 months (n = 7). Basal myocardial blood flow in group 1 to 12 months (1.86+/-1.01 ml/min/g) exceeded (p <0.05) that of group 13 to 34 months (1.17+/-0.73) and group > or = 37 months (0.98+/-0.34). In group 13 to 34 months, basal myocardial blood flow and maximal dilator capacity (minimal coronary vascular resistance with adenosine 36+/-12 mm Hg/ml/min/g) were comparable to that of normal volunteers (1.01+/-0.20 and 37+/-, respectively). In group > or = 37 months, maximal flow response to adenosine was reduced (2.54+/-1.25 vs 3.16+/-0.52, respectively, p = 0.06). Maximal dilator capacity in group > or = 37 months (60+/-34) was impaired versus group 1 to 12 months (36+/-10) and group 13 to 34 months (36+/-12; both p <0.05) as well as normals (37+/-9, p <0.05). During the first year after cardiac transplantation basal myocardial blood flow is elevated out of proportion to external determinants of myocardial oxygen demand, but maximal dilator capacity of the coronary microcirculation is normal. Between 1 and 3 years both basal myocardial blood flow and microvascular function tend to normalize. After 3 years, although basal myocardial blood flow is normal, microvascular dilator capacity is impaired. PMID- 9856924 TI - Comparison of left ventricular function during interval versus steady-state exercise training in patients with chronic congestive heart failure. AB - This study sought to assess the safety of interval exercise training in patients with chronic congestive heart failure (CHF) with respect to left ventricular (LV) function. For effective rehabilitation in CHF, both aerobic capacity and muscle strength need to be improved. We have previously demonstrated in both coronary artery bypass surgery and patients with CHF that interval exercise training (IET) offers advantages over steady-state exercise training (SSET). However, because LV function during IET has not yet been studied, the safety of this method in CHF remains unclear. To assess LV function during IET and SSET, at the same average power output, 11 patients with stable CHF were compared with 9 stable coronary patients with minimal LV dysfunction (control group). Using first-pass radionuclide ventriculography, changes in LV function were assessed during work versus recovery phases, at temporally matched times between the fifth and sixteenth minute of IET and SSET. In CHF during IET, there were no significant variations in the parameters measured during work and/or recovery phases. During the course of both IET and SSET, there was a significant increase in LV ejection fraction (5 vs 4 U; p <0.05 each), accompanied by increased heart rate (6 vs 8 beats/min; p <0.05 each) and cardiac output (2.4 vs 1.8 L/min; p <0.01 and p <0.05). In CHF, the magnitude of change in LV ejection fraction during IET was similar to that seen in controls. Both LV ejection fraction and the clinical status in patients with CHF remained stable during IET. Because IET appears to be as safe as SSET with respect to LV function, IET can be recommended for exercise training in CHF to apply higher peripheral exercise stimuli and with no greater LV stress than during SSET. PMID- 9856925 TI - What does the left atrial v wave signify during balloon commissurotomy of mitral stenosis? AB - Left atrial v-wave amplitude has been associated with the presence and severity of chronic mitral regurgitation (MR) but it has not been evaluated for the detection of acute MR. We evaluated the left atrial v-wave amplitude of 205 consecutive patients with mitral stenosis immediately before and after stepwise, incremental balloon mitral commissurotomy to determine predictors of large v waves at baseline and an increase in v-wave amplitude after balloon commissurotomy. The sensitivity and specificity of an increase in v-wave amplitude for detecting worsening and severe MR were determined. A large v wave was present in 44% of patients before balloon commissurotomy and was predicted by age, mean left atrial pressure, mean transmitral gradient, mean pulmonary artery pressure, and angiographic severity of MR. There was a strong inverse correlation between v-wave amplitude and calculated left atrial compliance (r = -0.92). An increase in v-wave amplitude after balloon commissurotomy was associated with an increasing probability of worsening or severe MR. This indicator had a sensitivity, specificity, and positive and negative predictive values of 35%, 91%, 64%, 75%, respectively, for detecting any increase in MR. For the detection of severe MR, the sensitivity was 79%, specificity 89%, positive predictive value 42%, and negative predictive value 98%. Thus, left atrial v-wave amplitude reflects left atrial compliance and severity of mitral stenosis before balloon commissurotomy. An increase in v-wave amplitude is an insensitive but very specific indicator of worsening or severe MR during stepwise, incremental balloon mitral commissurotomy. PMID- 9856926 TI - Use of pulsed Doppler tissue imaging to assess regional left ventricular diastolic dysfunction in hypertrophic cardiomyopathy. AB - In this study, regional diastolic patterns and their relations with transmitral Doppler inflow were investigated in hypertrophic cardiomyopathy (HC) by pulsed Doppler tissue imaging (DTI). Doppler echocardiography and DTI of basal septum and lateral wall (apical 4-chamber view) were performed in 20 patients (15 men and 5 women) with HC and in 10 healthy subjects (7 men and 3 women). Diabetes, hypertension, coronary artery and valvular disease, mitral regurgitation, New York Heart Association functional classes III to IV, sinus tachycardia, atrial fibrillation, and inadequate echocardiograms were exclusion criteria. Peak velocity and time-velocity integral of early and late waves and their ratios, and deceleration and isovolumic relaxation times were determined by standard Doppler and by DTI at the septal and lateral wall levels. The 2 groups were comparable for age, heart rate, blood pressure, and ejection fraction. Transmitral peak velocity and time-velocity integral E/A ratios were reduced (both p <0.05) and deceleration and isovolumic relaxation times prolonged (both p <0.00001) in HC. Septal DTI showed lower peak velocity and time-velocity integral e/a ratios (p <0.00001 and p <0.001, respectively) and lengthened regional deceleration (p <0.01) and isovolumic (p <0.001) relaxation times. DTI of the lateral wall showed a prolongation of deceleration and isovolumic relaxation times (both p <0.01). By dividing HC according to transmitral E/A, 8 patients with E/A <1 had lower DTI septal e/a ratio (p <0.01) and prolonged septal deceleration and isovolumic relaxation times (both p <0.01) but no changes in DTI pattern of lateral wall than 12 patients with E/A > 1. In conclusion, DTI is useful and complementary to standard Doppler imaging to characterize diastolic properties in HC, reflecting a typical pattern of intramyocardial impaired relaxation at the level of hypertrophied septum and also providing information about the degree of this regional impairment. The lateral wall presents minor changes in diastolic times, which indicate how diastolic asynchrony is not confined to the hypertrophied segment in HC. PMID- 9856927 TI - Diagnosis of arrhythmogenic right ventricular cardiomyopathy by fourier analysis of gated blood pool single-photon emission tomography. AB - To evaluate the diagnostic performance of Fourier phase analysis of gated blood pool single-photon emission computed tomography (GBP SPECT) in arrhythmogenic right ventricular (RV) cardiomyopathy, 18 patients with confirmed arrhythmogenic RV cardiomyopathy underwent GBP SPECT and x-ray cineangiography. Results were compared with data obtained with GBP SPECT in 10 control subjects. This 3 dimensional method demonstrated good correlation with cineangiography for measurements of RV enlargement and extent of the disease; RV and left ventricular segments were analyzed with the same accuracy. Tomographic abnormalities were significant decreased RV ejection fraction, RV dilatation, nonsynchronized contraction of the ventricles, increased RV contraction dispersion, presence of segmental RV wall motion disorders and/or phase delays, and occasionally regional left ventricular abnormalities. RV-delayed phase areas were always present in our population. A scoring system with RV criteria was proposed to diagnose RV disease. Because Fourier analysis of GBP SPECT provides ventricular morphologic information for the right ventricle with the same accuracy as for the left ventricle, it may replace planar radionuclide studies. Therefore, this method is helpful in patients with a strong clinical suspicion of arrhythmogenic RV cardiomyopathy, and should be used as a screening method before right ventriculography. PMID- 9856928 TI - Transcatheter closure of atrial septal defect and patent foramen ovale with ASDOS device (a multi-institutional European trial). AB - A clinical trial was conducted to assess the feasibility, safety, and efficacy of the atrial septal defect (ASD) occlusion system for transcatheter closure of secundum ASD and patent foramen ovale (PFO) after episodes of cerebral embolism. Occlusion was attempted in 200 patients aged 1 to 74 years (mean 32). The procedure failed in 26 patients (13%); the device was retrieved through a catheter in 20 and through surgery in 6 patients. Procedure-related complications necessitating surgical removal of the device included device embolization in 2, device entrapment within the Chiari network in 1, frame fracture in 1, and perforation of atrial wall in 2. All 6 patients experienced an uneventful postoperative course. An additional 11 patients (6%) underwent surgical removal of the device during follow-up. There were 163 patients (81%) with an implanted ASD occlusion system at follow-up of from 6 to 36 months (mean 17). Thrombus formation around the device was detected by transesophageal echocardiography in 9 patients 1 to 4 weeks after implantation. One of these patients (who had a coagulation factor XII deficiency) suffered a cerebral thromboembolism. Late atrial wall perforation (5, 6, and 8 months after implantation) occurred in 3 adult patients. Infectious endocarditis developed in 2 adult patients (1%). No late device embolization and no atrioventricular valve injury occurred. An asymptomatic device frame fracture was found in 14% and frame deformity in 4% of all patients during the follow-up period of >230 patient-years. Immediately after closure, a moderate/large residual shunt remained in 8% and a small shunt in 29% of patients. After 1 year, a moderate/large shunt was present in 2% and a small one in 26% of patients. During a total follow-up of 49 patient-years, only 1 of 46 patients with PFO had a transient neurologic event after the closure. The study indicates that patients with centrally situated secundum ASD and those with PFO after cerebral embolism can be treated with this system with a high success rate and an acceptable morbidity. PMID- 9856929 TI - Evaluation of left ventricular early diastolic performance by color tissue Doppler imaging of the mitral annulus. AB - A noninvasive assessment of left ventricular (LV) diastolic performance by tissue Doppler imaging was performed in 56 patients (8 patients with atypical chest pain, 42 with coronary artery disease with a previous myocardial infarction, and 6 without a previous myocardial infarction) who underwent cardiac catheterization. Mitral annular velocity (MAV) during early ventricular diastole was obtained by M-mode color tissue Doppler imaging at the posterior corner of the mitral annulus. In each patient, the negative peak of the first derivative of LV pressure decay (peak -dP/dt) and a time constant of LV relaxation (tau) were calculated from the LV pressure waves obtained by a catheter-tip micromanometer. LV end-systolic volume index was measured from contrast left ventriculography. MAV during early diastole was significantly correlated with tau (r = -0.73, p <0.001), peak -dP/dt (r = 0.58, p <0.001), and LV end-systolic volume index (r = 0.63, p <0.001). On multivariate regression analysis with MAV during early diastole, tau and LV end-systolic volume index were selected as prime determinants (r = 0.80, p <0.001). These findings suggest that MAV during early diastole has a direct relation to LV elastic recoil as well as to LV relaxation. MAV during early diastole gives important information regarding LV behavior in late systole to early diastole where LV early diastolic performance is determined. PMID- 9856930 TI - Coronary stenting of bifurcation lesions using "T" or "reverse Y" configuration with Wiktor stent. AB - The purpose of this study was to determine the immediate and long-term angiographic and clinical results of coronary stenting of bifurcation lesions using the "T" or "reverse Y" configuration with a Wiktor GX stent. This series demonstrates that stents can be placed with a high rate of success in coronary bifurcation lesions and the techniques described will overcome the potential limitations of coronary angioplasty in such lesions. PMID- 9856931 TI - Randomized, prospective comparison of four burst pacing algorithms for spontaneous ventricular tachycardia. AB - "Less aggressive" burst stimulation is more effective in terminating spontaneous monomorphic ventricular tachycardia with a lesser acceleration rate. Higher ventricular tachycardia cycle length and use of 91% coupling interval were independent predictors for pacing termination. PMID- 9856932 TI - Chronotropic response, safety, and accuracy of dobutamine stress echocardiography in patients with atrial fibrillation and known or suspected coronary artery disease. AB - Ninety-two consecutive patients with atrial fibrillation (AF) who underwent dobutamine stress echocardiography were compared with a control group of patients in sinus rhythm matched for age, sex, and resting heart rate. Patients with AF had an increased chronotropic response to dobutamine, but there were no adverse effects and no evidence that the lower doses of dobutamine typically given to patients with AF were insufficient to induce ischemia. PMID- 9856933 TI - Comparison of echocardiographic acoustic quantification with off-line manual tracing for determining left ventricular volume and ejection fraction in pediatric patients. AB - Echocardiographic measurement of left ventricular systolic and diastolic volume and ejection fraction in pediatric patients by acoustic quantification using automated border methods compares well with measurements done by manual trace. The time necessary for completion of measurements was similar for the two methods. PMID- 9856934 TI - Relation between the number of image planes and the accuracy of three-dimensional echocardiography for measuring left ventricular volumes and ejection fraction. AB - The relation between accuracy of 3-dimensional echocardiography (3DE) in determining left ventricular end-diastolic volume, end-systolic volume, and ejection fraction (compared with magnetic resonance imaging) and the number of component planes used for 3DE ventricular reconstruction was evaluated in 41 adult subjects with normal (n = 24) and abnormal (n = 17) left ventricles. Accuracy and confidence of 3DE gradually increased with use of additional component planes, so that > or = 10 planes from both parasternal and apical windows provided 3DE reconstructions that accurately predict magnetic resonance imaging-measured left ventricular volumes and ejection fraction with confidence. PMID- 9856935 TI - Location of stents in the left anterior descending coronary artery using three dimensionally acquired, two dimensionally displayed transthoracic echocardiography. AB - The location of left anterior descending arterial stents was documented in 10 of 13 patients using 3-dimensionally acquired, 2-dimensionally displayed transthoracic echocardiography. The echocardiographic appearance of stents was characterized, and the location of the proximal and mid-left anterior descending artery was objectively documented. PMID- 9856936 TI - Carbon in the universe. AB - Carbon is a major player in the evolutionary scheme of the universe because of its abundance and its ability to form complex species. It is also a key element in the evolution of prebiotic molecules. The different forms of cosmic carbon are reviewed ranging from carbon atoms and carbon-bearing molecules to complex, solid state, carbonaceous structures. The current state of knowledge is assessed on the observational and laboratory fronts. Fundamental astrophysical implications are examined as well as the impact of these studies on the hitherto poorly understood physical and chemical properties of carbon materials in space. PMID- 9856937 TI - Siderophore-mediated iron transport: crystal structure of FhuA with bound lipopolysaccharide. AB - FhuA, the receptor for ferrichrome-iron in Escherichia coli, is a member of a family of integral outer membrane proteins, which, together with the energy transducing protein TonB, mediate the active transport of ferric siderophores across the outer membrane of Gram-negative bacteria. The three-dimensional structure of FhuA is presented here in two conformations: with and without ferrichrome-iron at resolutions of 2.7 and 2.5 angstroms, respectively. FhuA is a beta barrel composed of 22 antiparallel beta strands. In contrast to the typical trimeric arrangement found in porins, FhuA is monomeric. Located within the beta barrel is a structurally distinct domain, the "cork," which mainly consists of a four-stranded beta sheet and four short alpha helices. A single lipopolysaccharide molecule is noncovalently associated with the membrane embedded region of the protein. Upon binding of ferrichrome-iron, conformational changes are transduced to the periplasmic pocket of FhuA, signaling the ligand loaded status of the receptor. Sequence homologies and mutagenesis data are used to propose a structural mechanism for TonB-dependent siderophore-mediated transport across the outer membrane. PMID- 9856938 TI - Structure of the MscL homolog from Mycobacterium tuberculosis: a gated mechanosensitive ion channel. AB - Mechanosensitive ion channels play a critical role in transducing physical stresses at the cell membrane into an electrochemical response. The MscL family of large-conductance mechanosensitive channels is widely distributed among prokaryotes and may participate in the regulation of osmotic pressure changes within the cell. In an effort to better understand the structural basis for the function of these channels, the structure of the MscL homolog from Mycobacterium tuberculosis was determined by x-ray crystallography to 3.5 angstroms resolution. This channel is organized as a homopentamer, with each subunit containing two transmembrane alpha helices and a third cytoplasmic alpha helix. From the extracellular side, a water-filled opening approximately 18 angstroms in diameter leads into a pore lined with hydrophilic residues which narrows at the cytoplasmic side to an occluded hydrophobic apex that may act as the channel gate. This structure may serve as a model for other mechanosensitive channels, as well as the broader class of pentameric ligand-gated ion channels exemplified by the nicotinic acetylcholine receptor. PMID- 9856940 TI - A free-fall determination of the newtonian constant of gravity AB - Recent determinations of the Newtonian constant of gravity have produced values that differ by nearly 40 times their individual error estimates (more than 0.5%). In an attempt to help resolve this situation, an experiment that uses the gravity field of a one-half metric ton source mass to perturb the trajectory of a free falling mass and laser interferometry to track the falling object was performed. This experiment does not suspend the test mass from a support system. It is therefore free of many systematic errors associated with supports. The measured value was G = (6.6873 +/- 0. 0094) x 10(-11) m3 kg-1 sec-2. PMID- 9856939 TI - Regulation of polar auxin transport by AtPIN1 in Arabidopsis vascular tissue. AB - Polar auxin transport controls multiple developmental processes in plants, including the formation of vascular tissue. Mutations affecting the PIN-FORMED (PIN1) gene diminish polar auxin transport in Arabidopsis thaliana inflorescence axes. The AtPIN1gene was found to encode a 67-kilodalton protein with similarity to bacterial and eukaryotic carrier proteins, and the AtPIN1 protein was detected at the basal end of auxin transport-competent cells in vascular tissue. AtPIN1 may act as a transmembrane component of the auxin efflux carrier. PMID- 9856941 TI - Abrupt climate oscillations during the last deglaciation in central north america AB - Evidence from stable isotopes and a variety of proxies from two Ontario lakes demonstrate that many of the late glacial-to-early Holocene events that are well known from the North Atlantic seaboard, such as the Gerzensee-Killarney Oscillation (also known as the Intra-Allerod Cold Period), Younger Dryas, and Preboreal Oscillation, also occurred in central North America. These results thus imply that climatic forcing acted in the same manner in both regions and that atmospheric circulation played an important role in the propagation of these events. PMID- 9856942 TI - The percolation phase transition in sea Ice AB - Sea ice exhibits a marked transition in its fluid transport properties at a critical brine volume fraction pc of about 5 percent, or temperature Tc of about 5 degreesC for salinity of 5 parts per thousand. For temperatures warmer than Tc, brine carrying heat and nutrients can move through the ice, whereas for colder temperatures the ice is impermeable. This transition plays a key role in the geophysics, biology, and remote sensing of sea ice. Percolation theory can be used to understand this critical behavior of transport in sea ice. The similarity of sea ice microstructure to compressed powders is used to theoretically predict pc of about 5 percent. PMID- 9856943 TI - Evidence for extreme climatic warmth from late cretaceous arctic vertebrates AB - A Late Cretaceous (92 to 86 million years ago) vertebrate assemblage from the high Canadian Arctic (Axel Heiberg Island) implies that polar climates were warm (mean annual temperature exceeding 14 degreesC) rather than near freezing. The assemblage includes large (2.4 meters long) champsosaurs, which are extinct crocodilelike reptiles. Magmatism at six large igneous provinces at this time suggests that volcanic carbon dioxide emissions helped cause the global warmth. PMID- 9856944 TI - Hierarchically ordered oxides AB - Porous silica, niobia, and titania with three-dimensional structures patterned over multiple length scales were prepared by combining micromolding, polystyrene sphere templating, and cooperative assembly of inorganic sol-gel species with amphiphilic triblock copolymers. The resulting materials show hierarchical ordering over several discrete and tunable length scales ranging from 10 nanometers to several micrometers. The respective ordered structures can be independently modified by choosing different mold patterns, latex spheres, and block copolymers. The examples presented demonstrate the compositional and structural diversities that are possible with this simple approach. PMID- 9856945 TI - Atomic contributions to the optical rotation angle as a quantitative probe of molecular chirality. AB - Chiral molecules are characterized by a specific rotation angle, the angle through which plane-polarized light is rotated on passing through an enantiomerically enriched solution. Recent developments in methodology allow computation of both the sign and the magnitude of these rotation angles. However, a general strategy for assigning the individual contributions that atoms and functional groups make to the optical rotation angle and, more generally, to the molecular chirality has remained elusive. Here, a method to determine the atomic contributions to the optical rotation angle is reported. This approach links chemical structure with optical rotation angle and provides a quantitative measure of molecular asymmetry propagation from a center, axis, or plane of chirality. PMID- 9856946 TI - Behavioral state modulation of auditory activity in a vocal motor system. AB - Neurons of the song motor control nucleus robustus archistriatalis (RA) exhibited far weaker auditory responses in awake than in anesthetized zebra finches. Remarkably, sleep induced complex patterns of bursts in ongoing activity and uncovered vigorous auditory responses of RA neurons. Local injections of norepinephrine suggested that the changes in response strength occur through neuromodulatory control of the sensorimotor nucleus HVc, which projects to RA. Thus, motor access to auditory feedback, which zebra finches require for song learning and maintenance, may be regulated through neuromodulation. During sleep, the descending motor system may gain access to sensorimotor song memories represented as bursting patterns of activity. PMID- 9856947 TI - Microscale nutrient patches in planktonic habitats shown by chemotactic bacteria AB - Are nutrients available to microbial communities in micropatches long enough to influence growth and competition? And what are the sources of such patches? To answer these questions, the swimming behavior of chemotactic bacteria in seawater samples was examined. Clusters of bacteria formed in conjunction with cell lysis and excretion by protozoa. These point sources of nutrients spread into spherical patches a few millimeters in diameter and sustained swarms of bacteria for about 10 minutes. Within that time, a large proportion of the nutrients was encountered by bacteria, chemotactic and nonchemotactic alike. Chemotaxis is advantageous for bacteria using patches over a certain size. PMID- 9856948 TI - Prevention of population cycles by parasite removal. AB - The regular cyclic fluctuations in vertebrate numbers have intrigued scientists for more than 70 years, and yet the cause of such cycles has not been clearly demonstrated. Red grouse populations in Britain exhibit cyclic fluctuations in abundance, with periodic crashes. The hypothesis that these fluctuations are caused by the impact of a nematode parasite on host fecundity was tested by experimentally reducing parasite burdens in grouse. Treatment of the grouse population prevented population crashes, demonstrating that parasites were the cause of the cyclic fluctuations. PMID- 9856949 TI - Interleukin-13: central mediator of allergic asthma. AB - The worldwide incidence, morbidity, and mortality of allergic asthma are increasing. The pathophysiological features of allergic asthma are thought to result from the aberrant expansion of CD4(+) T cells producing the type 2 cytokines interleukin-4 (IL-4) and IL-5, although a necessary role for these cytokines in allergic asthma has not been demonstrable. The type 2 cytokine IL 13, which shares a receptor component and signaling pathways with IL-4, was found to be necessary and sufficient for the expression of allergic asthma. IL-13 induces the pathophysiological features of asthma in a manner that is independent of immunoglobulin E and eosinophils. Thus, IL-13 is critical to allergen-induced asthma but operates through mechanisms other than those that are classically implicated in allergic responses. PMID- 9856951 TI - Molecular basis of T cell inactivation by CTLA-4. AB - CTLA-4, a negative regulator of T cell function, was found to associate with the T cell receptor (TCR) complex zeta chain in primary T cells. The association of TCRzeta with CTLA-4, reconstituted in 293 transfectants, was enhanced by p56(lck) induced tyrosine phosphorylation. Coexpression of the CTLA-4-associated tyrosine phosphatase, SHP-2, resulted in dephosphorylation of TCRzeta bound to CTLA-4 and abolished the p56(lck)-inducible TCRzeta-CTLA-4 interaction. Thus, CTLA-4 inhibits TCR signal transduction by binding to TCRzeta and inhibiting tyrosine phosphorylation after T cell activation. These findings have broad implications for the negative regulation of T cell function and T cell tolerance. PMID- 9856950 TI - Requirement for IL-13 independently of IL-4 in experimental asthma. AB - The pathogenesis of asthma reflects, in part, the activity of T cell cytokines. Murine models support participation of interleukin-4 (IL-4) and the IL-4 receptor in asthma. Selective neutralization of IL-13, a cytokine related to IL-4 that also binds to the alpha chain of the IL-4 receptor, ameliorated the asthma phenotype, including airway hyperresponsiveness, eosinophil recruitment, and mucus overproduction. Administration of either IL-13 or IL-4 conferred an asthma like phenotype to nonimmunized T cell-deficient mice by an IL-4 receptor alpha chain-dependent pathway. This pathway may underlie the genetic associations of asthma with both the human 5q31 locus and the IL-4 receptor. PMID- 9856952 TI - A receptor/cytoskeletal movement triggered by costimulation during T cell activation. AB - During T cell activation, the engagement of costimulatory molecules is often crucial to the development of an effective immune response, but the mechanism by which this is achieved is not known. Here, it is shown that beads attached to the surface of a T cell translocate toward the interface shortly after the start of T cell activation. This movement appears to depend on myosin motor proteins and requires the engagement of the major costimulatory receptor pairs, B7-CD28 and ICAM-1-LFA-1. This suggests that the engagement of costimulatory receptors triggers an active accumulation of molecules at the interface of the T cell and the antigen-presenting cell, which then increases the overall amplitude and duration of T cell signaling. PMID- 9856953 TI - p53-independent role of MDM2 in TGF-beta1 resistance. AB - Transforming growth factor-beta (TGF-beta) inhibits cell proliferation, and acquisition of TGF-beta resistance has been linked to tumorigenesis. A genetic screen was performed to identify complementary DNAs that abrogated TGF-beta sensitivity in mink lung epithelial cells. Ectopic expression of murine double minute 2 rescued TGF-beta-induced growth arrest in a p53-independent manner by interference with retinoblastoma susceptibility gene product (Rb)/E2F function. In human breast tumor cells, increased MDM2 expression levels correlated with TGF beta resistance. Thus, MDM2 may confer TGF-beta resistance in a subset of tumors and may promote tumorigenesis by interference with two independent tumor suppressors, p53 and Rb. PMID- 9856954 TI - Regulation of cocaine reward by CREB. AB - Cocaine regulates the transcription factor CREB (adenosine 3', 5'-monophosphate response element binding protein) in rat nucleus accumbens, a brain region that is important for addiction. Overexpression of CREB in this region decreases the rewarding effects of cocaine and makes low doses of the drug aversive. Conversely, overexpression of a dominant-negative mutant CREB increases the rewarding effects of cocaine. Altered transcription of dynorphin likely contributes to these effects: Its expression is increased by overexpression of CREB and decreased by overexpression of mutant CREB. Moreover, blockade of kappa opioid receptors (on which dynorphin acts) antagonizes the negative effect of CREB on cocaine reward. These results identify an intracellular cascade culminating in gene expression-through which exposure to cocaine modifies subsequent responsiveness to the drug. PMID- 9856955 TI - A family of cAMP-binding proteins that directly activate Rap1. AB - cAMP (3',5' cyclic adenosine monophosphate) is a second messenger that in eukaryotic cells induces physiological responses ranging from growth, differentiation, and gene expression to secretion and neurotransmission. Most of these effects have been attributed to the binding of cAMP to cAMP-dependent protein kinase A (PKA). Here, a family of cAMP-binding proteins that are differentially distributed in the mammalian brain and body organs and that exhibit both cAMP-binding and guanine nucleotide exchange factor (GEF) domains is reported. These cAMP-regulated GEFs (cAMP-GEFs) bind cAMP and selectively activate the Ras superfamily guanine nucleotide binding protein Rap1A in a cAMP dependent but PKA-independent manner. Our findings suggest the need to reformulate concepts of cAMP-mediated signaling to include direct coupling to Ras superfamily signaling. PMID- 9856956 TI - A memorial to Robert Tiegerstedt: the centennial of renin discovery. PMID- 9856958 TI - Effect of activation and inhibition of the renin-angiotensin system on plasma PAI 1. AB - Increased plasma renin activity (PRA) has been associated with an increased risk of myocardial infarction (MI), whereas angiotensin-converting enzyme (ACE) inhibition appears to reduce the risk of recurrent MI in patients with left ventricular dysfunction. These observations may be partially explained by an interaction between the renin-angiotensin system (RAS) and fibrinolytic system. To test this hypothesis, we examined the effect of salt depletion on tissue-type plasminogen activator (tPA) antigen and plasminogen activator inhibitor-1 (PAI-1) activity and antigen in normotensive subjects in the presence and absence of quinapril (40 mg BID). Under low (10 mmol/d) and high (200 mmol/d) salt conditions there was significant diurnal variation in PAI-1 antigen and activity and tPA antigen. Morning (8 AM through 2 PM) PAI-1 antigen levels were significantly higher during low salt intake compared with high salt intake conditions (ANOVA, F=5.8, P=0.048). PAI-1 antigen correlated with aldosterone (r=0.56, P<10(-7)) during low salt intake. ACE inhibition significantly decreased 24-hour (ANOVA for 24 hours, F=6. 7, P=0.04) and morning (F=24, P=0.002) PAI-1 antigen and PAI-1 activity (F=6.48, P=0.038) but did not alter tPA antigen. Thus, the mean morning PAI-1 antigen concentration was significantly higher during low salt intake than during either high salt intake or low salt intake and concomitant ACE inhibition (22.7+/-4.6 versus 16. 1+/-3.3 and 16.3+/-3.7 ng/mL, respectively; P<0.05). This study provides evidence of a direct functional link between the RAS and fibrinolytic system in humans. The data suggest that ACE inhibition has the potential to reduce the incidence of thrombotic cardiovascular events by blunting the morning peak in PAI-1. PMID- 9856959 TI - Inverse relationship between fenfluramine-induced prolactin release and blood pressure in humans. AB - Although substantial evidence from experimental animals suggests that augmentation and reduction in serotonergic neurotransmission both affect arterial blood pressure (BP), it is unknown whether "tonic" central serotonergic activity is related to resting BP variability in humans. We tested this hypothesis in a community sample by evaluating the relationship between resting BP and a neuropharmacologic index of brain serotonergic activity (the fenfluramine challenge test). Subjects were 270 generally healthy men and women aged 25 to 60 years who were not receiving prescribed antihypertensive or psychotropic medications. The sample included 216 non-Hispanic whites and 47 blacks. Resting systolic BP ranged from 85 to 161 mm Hg and diastolic from 58 to 98 mm Hg. Each subject received 0.55 to 0.65 mg/kg D,L-fenfluramine hydrochloride, and the plasma prolactin concentration was measured over 3.5 hours. Analyses revealed a linear, inverse relationship between the maximum fenfluramine-induced prolactin rise and systolic and diastolic BP in whites: r=-0.36 and r=-0.29, respectively (P<0.001 for both). These relationships were not observed in the black participants. In whites, the prolactin response to fenfluramine remained a significant predictor of systolic and diastolic BPs in multivariate models including age, gender, body mass index, physical activity, smoking, and alcohol consumption (P135/85 mm Hg. These data reveal that in white but not black adults, fenfluramine-induced prolactin release correlates inversely with BP and may indicate a role of central serotonergic activity in the pathogenesis of hypertension. PMID- 9856957 TI - Chronic nitric oxide inhibition model six years on. PMID- 9856960 TI - Intracellular angiotensin II regulates the inward calcium current in cardiac myocytes. AB - -The influence of intracellular administration of angiotensin II (Ang II) on the inward calcium current (ICa) was investigated in single myocytes isolated from adult rat ventricle. Comparative studies were also made in ventricular cells of Golden hamsters. The ICa was measured in single cells using the whole-cell voltage clamp configuration. The results indicated that Ang II (10(-8) mmol/L) dialyzed into the rat myocytes reduced the peak ICa by 35+/-5.5% (n=20; P<0.05). Losartan (10(-7) mmol/L) added to the bath did not suppress the effects of Ang II, indicating that the peptide is acting intracellularly. Moreover, the intracellular dialysis of losartan (10(-6) mmol/L) or [Sar1Val5Ala8] Ang II (10( 6) mmol/L) did not change the effect of Ang II. Stimulation of ICa by exogenous cAMP or inhibition of protein kinase C did not alter the effect of Ang II on ICa. Zaprinast (100 micromol/L), an inhibitor of cGMP phosphodiesterase, when added to the bath solution increased appreciably the effect of Ang II on ICa (P<0.05). In ventricular myocytes of Golden hamsters, in which Ang II has a positive inotropic action, the intracellular administration of Ang II (10(-8) mmol/L) increased ICa by 36+/-2.4% (n=20; P>0.05). The effect of the peptide was not altered by the intracellular administration of losartan (10(-6) mmol/L), by [Sar1Val5Ala8] Ang II (10(-6) mmol/L), or by the inhibitor of protein kinase A. The inhibition of protein kinase C, however, prevented the effect of Ang II ICa in the hamster myocytes. The results particularly suggest that the activation of the cardiac renin-angiotensin system regulates ICa and myocardial contractility, an effect that varies with the species. PMID- 9856961 TI - Ambulatory pulse pressure: a potent predictor of total cardiovascular risk in hypertension. AB - A wide pulse pressure (PP) is a marker of increased artery stiffness and high cardiovascular (CV) risk. To investigate the prognostic value of ambulatory PP, which is currently unknown, we studied 2010 initially untreated subjects with uncomplicated essential hypertension (mean age, 51.7 years; 52% men). All subjects underwent baseline procedures including 24-hour noninvasive ambulatory blood pressure (BP) monitoring. The mean duration of follow-up was 3.8 years (range, 0 to 11 years), and CV morbidity and mortality were the outcome measures. There were 200 major CV events (2.61 per 100 person-years), 36 of which were fatal (0.47 per 100 person-years). In the 3 tertiles of the distribution of office PP, the rate of total CV events (per 100 persons per year) was 1.38, 2. 12, and 4.34, respectively, and that of fatal events was 0.12, 0.30, and 1.07 (log-rank test, both P<0.01). In the 3 tertiles of the distribution of average 24 hour PP, the rate of total CV events was 1.19, 1.81, and 4.92, and that of fatal events was 0.11, 0.17, and 1. 23 (log-rank test, both P<0.01). After controlling for several independent risk markers including white coat hypertension and nondipper status, we found that ambulatory PP was associated with the biggest reduction in the -2 log likelihood statistics for CV morbidity (P<0.05 versus office PP). In each of the 3 tertiles of office PP, CV morbidity and mortality increased from the first to the third tertile of average 24-hour ambulatory PP (log-rank test, all P<0.01). Age, left ventricular hypertrophy, and nondipper status were independent predictors of CV mortality, and the further predictive effect of ambulatory PP (P<0.001) was marginally but not significantly superior to that of office PP and average 24-hour systolic BP. We conclude that ambulatory PP is a potent risk marker in essential hypertension. CV morbidity is more closely predicted by ambulatory than by office PP even after control for multiple risk factors. PMID- 9856962 TI - Characteristics of 9194 patients with left ventricular hypertrophy: the LIFE study. Losartan Intervention For Endpoint Reduction in Hypertension. AB - -Losartan was the first available orally administered selective antagonist of the angiotensin II type 1 receptor developed for the treatment of hypertension. The Losartan Intervention For Endpoint (LIFE) Reduction in Hypertension Study is a double-blind, prospective, parallel group study designed to compare the effects of losartan with those of the beta-blocker atenolol on the reduction of cardiovascular morbidity and mortality. Patients with essential hypertension, aged between 55 and 80 years, and ECG-documented left ventricular hypertrophy (LVH) were included. Altogether, 9223 patients in Scandinavia, the United Kingdom, and the United States were randomized from June 1995 through April 1997, and 9194 remain after exclusion of a study center at which irregularities were discovered. This population of hypertensives (mean systolic/diastolic blood pressure, 174.4/97.8 mm Hg) with LVH comprises women (54.1%) and men, mostly retired from active work (mean age, 66.9 years), with a high prevalence of overweight (mean body mass index, 28.0 kg/m2), diabetes mellitus (12.3%), lipid disorders (18.0%), and symptoms or signs of coronary heart disease (15.1%). There were fewer current smokers (<17%) than in the general population, and approximately 7% were nonwhite. Almost 30% of participants had been untreated for at least 6 months when screened for the study. Only 1557 persons who entered the placebo run-in period of 14 days were excluded, predominantly because of sitting blood pressures above or below the predetermined range of 160-200/95-115 mm Hg and ECG-LVH criteria not met. By application of simple 12-lead ECG criteria for LVH (Cornell voltage QRS duration product formula plus Sokolow-Lyon voltage read by a core laboratory), hypertensive patients with LVH with an average 5-year coronary heart disease risk of 22.3% according to the Framingham score were identified. This population is now being treated (goal, <140/90 mm Hg) in adherence with the protocol for at least 4 years after final enrollment (ie, through April 2001) and until at least 1040 patients suffer myocardial infarction, stroke, or cardiovascular death. PMID- 9856963 TI - Blood pressure in Spain: distribution, awareness, control, and benefits of a reduction in average pressure. AB - Distribution of blood pressure (as per US Joint National Committee VI classification and staging criteria) plus awareness, treatment, and control of hypertension were studied in a representative Spanish population sample of 2021 persons (age range, 35 to 64 years). Pressure was determined in accordance with World Health Organization guidelines. A total of 45.1% of subjects were hypertensive (>/=140/>/=90 mm Hg or undergoing drug therapy); 12% had isolated systolic hypertension, and 8.7% had isolated diastolic hypertension. Pulse pressure was 48.7 mm Hg. Heart rate was 81.4 bpm in untreated hypertensives and 78.9 bpm in normotensives (P<0.05). A substantial proportion of the community burden of blood pressure was attributable to stage 1 (28.3% of subjects), the most frequent category of hypertension, and to the high-normal blood pressure group (17% of subjects). A percentage breakdown showed that among hypertensives, 44.5% were aware of their condition; of these, 71.9% were undergoing drug therapy, and of those being treated, only 15.5% were controlled (5% of hypertensives). Not only are these figures consistent with the fact that Spain has a higher cerebrovascular mortality than other countries such as the United States, but they represent a great potential for improvement, particularly among those groups registering relatively worse data (younger men, rural residents, and unskilled professionals). A decrease of only 1 to 4 mm Hg in average blood pressure could reduce the prevalence of hypertension in Spain by 12.4% to 15.4%. PMID- 9856964 TI - Reactive oxygen species are critical in the oleic acid-mediated mitogenic signaling pathway in vascular smooth muscle cells. AB - Obese hypertensive patients with cardiovascular risk factor clustering have increased plasma nonesterified fatty acid levels and are at high risk for atherosclerotic events. Our previous studies demonstrated that oleic acid induces a mitogenic response in rat aortic smooth muscle cells (RASMCs) through protein kinase C (PKC)- and extracellular signal-regulated kinase (ERK)-dependent pathways. In the present study we investigated the possibility that the generation of reactive oxygen species (ROS) constitutes a critical component of the oleic acid-induced mitogenic signaling pathway in RASMCs. We studied the effect(s) of oleic acid on the generation of ROS using the oxidant-sensitive fluoroprobe 2',7'-dichlorofluorescin diacetate. Relative fluorescence intensity and fluorescent images were obtained with laser confocal scanning microscopy from 1 to 5 minutes, since preliminary studies demonstrated that the peak fluorescence intensity occurred within 5 minutes. Oleic acid (100 micromol/L) induced a time dependent increase of cell fluorescence that was >8-fold of that seen in control cells at 5 minutes. This was blocked by catalase, which suggests that H2O2 was the principal ROS. The oleic acid-induced increases in H2O2 were blocked when PKC was inhibited with the use of bisindolylmaleimide and when PKC activity was downregulated by exposing RASMCs to phorbol 12-myristate 13-acetate for 24 hours. Stearic and elaidic acids, which are weak PKC activators, did not significantly increase H2O2 production. The increase of H2O2 in response to oleic acid was inhibited by the antioxidant N-acetylcysteine. N-Acetylcysteine also completely blocked ERK activation and the increase of thymidine incorporation in response to oleic acid. The data suggest that generation of H2O2 in RASMCs exposed to oleic acid is PKC dependent. Moreover, H2O2 production emerges as a critical intermediary event in the oleic acid-mediated mitogenic signaling pathway between the activation of PKC and ERK. These observations raise the possibility that the elevated plasma nonesterified fatty acids, including oleic acid, in obese hypertensive patients contribute to vascular growth and remodeling by a PKC dependent mechanism to generate ROS that subsequently activate ERK. PMID- 9856965 TI - Felodipine inhibits free-radical production by cytokines and glucose in human smooth muscle cells. AB - An imbalance between nitric oxide (NO) and superoxide is importantly involved in the pathogenesis of vascular disease. Inflammatory stimuli and risk factors contribute to these alterations. Calcium antagonists and angiotensin-converting enzyme inhibitors are commonly used cardiovascular drugs. To clarify the effect of felodipine and ramiprilat on the balance of these free radicals, we stimulated human aortic smooth muscle cells (HASCs) with cytokines (human interleukin-1beta, tumor necrosis factor-alpha, lipopolysaccharide, and/or interferon-gamma) or high glucose in the presence and absence of these compounds. Felodipine, but not ramiprilat, concentration-dependently inhibited cytokine-induced NO production and NO synthase (NOS) mRNA induction. The antioxidant N-acetylcysteine also inhibited cytokine-induced NO production and induction of inducible NOS mRNA. Moreover, felodipine inhibited cytokine-induced superoxide production both in the presence and absence of an NOS inhibitor, suggesting that it acted as a superoxide scavenger and not as an inhibitor of inducible NOS induction. High glucose treatment (22 mmol/L for 48 hours) also significantly increased superoxide production in HASCs, and this increase was inhibited in a concentration-dependent manner by felodipine but not by ramiprilat. These results suggest that felodipine may exert vascular protective effects by suppressing free radical generation in human smooth muscle cells during activation of inflammatory mechanisms and diabetic conditions. PMID- 9856966 TI - Basal and stimulated sympathetic responses after epinephrine: no evidence of augmented responses. AB - Delayed facilitation of norepinephrine release through the action of epinephrine (NE) at presynaptic beta-adrenoceptors has been postulated to account for the delayed hemodynamic effects of epinephrine and to be a mechanism causally related to the development of hypertension. To determine whether a short-term increase in epinephrine concentrations resulted in subsequent facilitation of sympathetic responses, 9 healthy subjects (age, 21+/-0.9 years) were studied at rest and during physiological stress on 2 occasions when they received an infusion of either saline or epinephrine (20 ng/kg per minute) in random order. Heart rate, blood pressure, forearm blood flow, epinephrine concentrations, and NE spillover were measured at rest, during mental stress (Stroop test), and during a cold pressor test. Measurements were performed before, during the 1-hour infusion of epinephrine or placebo, and 1 hour after the infusion. A radioisotope dilution method was used to measure NE spillover. Hemodynamic measurements and NE spillover were increased during the infusion of epinephrine, but 1 hour after discontinuation of epinephrine there was no significant augmentation of hemodynamic or sympathetic responses. NE spillover 1 hour after saline or epinephrine infusion was similar (0.85+/-0.2 versus 0. 87+/-0.2 microg/min; P=0.92). In addition, there was no delayed facilitation of stress-induced hemodynamic or NE responses after epinephrine. These findings do not support the hypothesis that epinephrine results in delayed facilitation of NE release. PMID- 9856968 TI - Both brain angiotensin II and "ouabain" contribute to sympathoexcitation and hypertension in Dahl S rats on high salt intake. AB - -Dahl salt-sensitive (Dahl S) and salt-resistant (Dahl R) rats from 5 to 9 weeks of age received a regular or high salt diet and concomitantly an intracerebroventricular infusion of the angiotensin type 1 blocker losartan (1 mg. kg-1. d-1) or antibody Fab fragments, which bind ouabain and related steroids with high affinity, or gamma-globulins as control (200 microg/d for both). At 9 weeks of age, blood pressure (BP), heart rate (HR), central venous pressure, and renal sympathetic nerve activity were recorded in conscious rats at rest and in response to air stress and to intracerebroventricular alpha2-agonist guanabenz (50 microg) and ouabain (0.5 microg). Baroreflex function was assessed by acute volume expansion with intravenous 5% dextrose and ramp changes of BP by +/-50 mm Hg induced by intravenous phenylephrine and sodium nitroprusside. In Dahl S but not R rats, high salt significantly increased BP and HR; enhanced BP, HR, and renal sympathetic nerve activity responses to air stress and guanabenz; and attenuated cardiopulmonary and arterial baroreflex control of renal sympathetic nerve activity and HR. Both losartan and Fab fragments prevented these responses to high salt to a similar extent in Dahl S rats but had no effect in Dahl R rats on high salt. Sympathoexcitatory responses to ouabain were attenuated in Dahl S on high versus regular salt and were abolished in Dahl R or S treated with losartan or Fab fragments. Consistent with previous studies in SHR, the present data indicate that in Dahl S on high salt, both brain "ouabain" and angiotensin II contribute to decreased sympathoinhibition and increased sympathoexcitation, impairment of baroreflex, and therefore hypertension. PMID- 9856967 TI - I1-imidazoline agonist moxonidine decreases sympathetic nerve activity and blood pressure in hypertensives. AB - Moxonidine is an I1-imidazoline receptor agonist that reduces blood pressure in hypertensives. Experimental data suggest that moxonidine inhibits central sympathetic activity. However, whether such a mechanism is involved in vivo in humans is still unclear. We investigated the effects of 0.4 mg moxonidine orally on muscle sympathetic nerve activity and heart rate in an open study in 8 healthy volunteers. Furthermore, we studied the effects of 0.4 mg moxonidine on muscle sympathetic nerve activity, heart rate, blood pressure, 24-hour blood pressure profile, and hormone plasma levels in 25 untreated hypertensives in a double blind, placebo-controlled study. Moxonidine decreased muscle sympathetic nerve activity in both healthy volunteers (P<0.05 versus baseline) and hypertensives (P<0.02 versus placebo). Plasma norepinephrine also decreased (P<0. 01), whereas plasma epinephrine and renin levels did not change (P=NS). Furthermore, moxonidine decreased systolic (P<0.0001) and diastolic (P<0.001) blood pressure. Heart rate decreased after moxonidine in healthy subjects (P<0.05); in hypertensives, heart rate decreased during the night hours (P<0.05) but not during daytime (P=NS). Plasma levels of LDL, HDL, and total cholesterol were not influenced by the drug (P=NS). Moxonidine decreases systolic and diastolic blood pressure by inhibiting central nervous sympathetic activity. This makes this new drug suitable for the treatment of human hypertension and possibly for other cardiovascular diseases with increased sympathetic nerve activity, ie, ischemic heart disease and heart failure. PMID- 9856969 TI - Cardiovascular effects of nitric oxide and adenosine in the nucleus tractus solitarii of rats. AB - It has been shown that nitric oxide (NO) is synthesized in the central nervous system as well as in vascular endothelial cells. We recently reported that NO was involved in central cardiovascular regulation, modulated the baroreflex, and was involved in a reciprocal release with excitatory amino acids in the nucleus tractus solitarii (NTS) of rats. We also reported previously that adenosine increased the release of glutamate in the NTS. The purpose of the present study was to investigate the possible interaction of NO and adenosine in the NTS. Male Sprague-Dawley rats were anesthetized with urethane, and blood pressure was monitored intra-arterially. Unilateral microinjection of L-arginine (3.3 nmol/60 nL) into the NTS produced decreases in blood pressure and heart rate. Microinjection of adenosine (2.3 nmol/60 nL) also produced depressive and bradycardic effects. These cardiovascular effects were attenuated by prior administration of the specific adenosine receptor antagonist DPSPX (0.92 nmol). Similarly, prior administration of NO synthase inhibitor NG-monomethyl-L-arginine or NG-nitro-L-arginine methyl ester significantly attenuated the depressive and bradycardic effects of adenosine. These results demonstrate a reciprocal attenuation of adenosine receptor antagonist and NO synthase inhibitor on L arginine and adenosine responses, respectively, in the NTS and implicate an interaction between NO and adenosine in central cardiovascular regulation. PMID- 9856970 TI - Baroreflex control of sympathetic nerve activity and heart rate in obstructive sleep apnea. AB - -Patients with obstructive sleep apnea are at increased risk for hypertension. The mechanisms underlying this increased risk are not known. We tested the hypothesis that obstructive sleep apnea, independent of factors such as hypertension, obesity, and age, is characterized by impairment of baroreflex sensitivity. We measured muscle sympathetic nerve activity (MSNA) and heart rate responses to activation and deactivation of baroreceptors in newly diagnosed, never treated, normotensive patients with obstructive sleep apnea. These responses were compared with those obtained in healthy control subjects closely matched for age, body mass index, and blood pressure. Heart rate and MSNA changes during infusion of phenylephrine (baroreceptor activation) were similar in the control subjects and patients with sleep apnea. Infusion of nitroprusside (baroreceptor deactivation) elicited similar decreases in mean arterial pressure (MAP) but lesser MSNA increases in patients with sleep apnea than in control subjects. Calculation of DeltaMSNA/DeltaMAP ratio revealed that baroreflex regulation of sympathetic activity for similar blood pressure changes was diminished in patients with sleep apnea in comparison to normal control subjects (P=0.01). However, increases in heart rate during nitroprusside infusion were comparable in both groups. Sympathetic, blood pressure and heart rate responses to the cold pressor test were also similar in the 2 groups. Our results indicate that normotensive patients with sleep apnea have a selective impairment of the sympathetic response to baroreceptor deactivation but not to baroreceptor activation or to the cold pressor test. The impairment of baroreflex sympathetic modulation in patients with sleep apnea is not accompanied by any impairment of baroreflex control of heart rate. PMID- 9856971 TI - Alpha1-adrenergic plus angiotensin receptor blockade reduces atherosclerosis in apolipoprotein E-deficient mice. AB - -We have used the apolipoprotein E (apoE)-deficient mouse model to determine whether both the angiotensin II type 1 (AT1) and the alpha1-adrenergic receptors influence arteriosclerotic changes in this hyperlipidemic animal model. Mice were treated with antihypertensive drugs beginning at 9 weeks of age, and aortic atherosclerosis was measured after 12 weeks of treatment. Systolic blood pressure in the untreated apoE-deficient mouse averaged 104 mm Hg throughout the treatment period. Prazosin at a dose of 7.5 mg. kg-1. d-1 was ineffective in attenuating atherosclerosis and did not significantly reduce blood pressure. Losartan, at dosages of either 20 or 30 mg. kg-1. d-1, also did not influence atherosclerosis and had only a slight blood pressure-lowering effect. However, combined treatment with both prazosin and losartan markedly reduced atherosclerotic lesion development from an average lesion size per section of 2.6 to 1.5x10(5) microm2 (P<0.001) and significantly reduced blood pressure to 85+/-5 mm Hg. Treatment with NG-nitro-L-arginine methyl ester (40 mg. kg-1. d-1) produced significant elevations of blood pressure (127+/-3.8 mm Hg) but had no effect on the development of atherosclerosis. None of the treatments used affected plasma cholesterol throughout the 12-week period. These studies suggest that the vascular changes associated with atherosclerosis are influenced by a combination of AT1 and alpha1-adrenergic receptor activation. PMID- 9856972 TI - Dopamine D1 receptor and protein kinase C isoforms in spontaneously hypertensive rats. AB - -Dopamine, via D1-like receptors, stimulates the activity of both protein kinase A (PKA) and protein kinase C (PKC), which results in inhibition of renal sodium transport. Since D1-like receptors differentially regulate sodium transport in normotensive and hypertensive rats, they may also differentially regulate PKC expression in these rat strains. Thus, 2 different D1-like agonists (fenoldopam or SKF 38393) were infused into the renal artery of anesthetized normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) (n=5 to 6/drug/strain). Ten or 60 minutes after starting the D1-like agonist infusion, both the infused kidney and the noninfused kidney that served as control were prepared for analysis. The D1-like agonists produced a greater diuresis and natriuresis and inhibited Na+,K+-ATPase activity in proximal tubule (PT) and medullary thick ascending limb (mTAL) to a greater extent in WKY (Delta20+/-1%) than in SHR (Delta7+/-1%, P<0.001). D1-like agonists had no effect on PKC-alpha or PKC-lambda expression in either membrane or cytosol but increased PKC-theta expression in PT in both WKY and SHR at 10 minutes but not at 60 minutes. However, membranous PKC-delta expression in PT and mTAL decreased in WKY but increased in SHR with either 10 or 60 minutes of D1-like agonist infusion. D1 like agonists also decreased membranous PKC-zeta expression in PT and mTAL in WKY but increased it in PT but not in mTAL in SHR. We conclude that there is differential regulation of PKC isoform expression by D1-like agonists that inhibits membranous PKC-delta and PKC-zeta in WKY but stimulates them in SHR; this effect in SHR is similar to the stimulatory effect of norepinephrine and angiotensin II and may be a mechanism for their differential effects on sodium transport. PMID- 9856973 TI - Bromocriptine regulates angiotensin II response on sodium pump in proximal tubules. AB - -Dopamine and angiotensin II (Ang II) receptors have been reported to exhibit an interaction in renal proximal tubules. The present study was designed to investigate the regulation by a D2-like dopamine receptor of Ang II-mediated stimulation of Na,K-ATPase activity in the renal proximal tubules. Ang II (10( 13) to 10(-9) mol/L) stimulated Na,K-ATPase activity in the proximal tubules that was completely abolished when the tubules were pretreated with the D2-like receptor agonist bromocriptine (1 micromol/L) for 30 minutes. The effect of bromocriptine on Ang II response was prevented by domperidone (1 micromol/L), a D2-like dopamine receptor antagonist. Similarly, the inhibition of forskolin (1 micromol/L)-induced cAMP accumulation caused by Ang II (10 pmol/L) was also abolished in bromocriptine-pretreated tubules. Basal and forskolin-stimulated cAMP was not significantly different in bromocriptine-treated tubules compared with the control. [3H]-Ang II binding sites (angiotensin type 1 [AT1] receptors) were reduced by approximately 65% in bromocriptine-treated proximal tubules, a result that was further substantiated by Western blot analysis revealing a 50% decrease in AT1 receptors in bromocriptine-treated tubules compared with the control. Western blot analysis of G proteins revealed a 2-fold increase in Gsalpha and a 20% decrease in Gialpha1 and Gialpha2 in the bromocriptine-treated proximal tubules. Bromocriptine (1 micromol/L) alone stimulated Na,K-ATPase activity during the first 30 minutes of incubation, and thereafter the stimulation fell to the basal level. Similarly, bromocriptine-mediated inhibition of cAMP lasted only up to 20 minutes. The data suggest that preactivation of D2 like dopamine receptors abolishes Ang II-mediated stimulation of Na,K-ATPase activity and inhibition of cAMP accumulation. This phenomenon may be a consequence of a decrease in AT1 receptors and alterations in G protein levels in the proximal tubules. We propose that such a regulation of Ang II response by bromocriptine is the result of heterologous desensitization of the D2-like receptor system. PMID- 9856974 TI - Biophysical signals underlying myogenic responses in rat interlobular artery. AB - -To assess cellular mechanisms mediating myogenic responses of interlobular artery (ILA), experiments were performed with the use of isolated perfused hydronephrotic kidneys. ILAs were divided into 3 groups according to their basal diameters: proximal (>60 microm), intermediate (40 to 60 microm), and distal (<40 microm) ILAs. Myogenic responses were obtained by stepwise increase in perfusion pressure. Greater myogenic responsiveness was observed in ILAs with smaller diameters. Diltiazem (10 micromol/L) inhibited myogenic responses of all segments of ILAs. Furthermore, gadolinium (10 micromol/L), a mechanosensitive cation channel blocker, abolished myogenic responses of distal but not proximal ILA. In contrast, 2-nitro-4-carboxyphenyl-N, N-diphenyl-carbamate (200 micromol/L), an inhibitor of phospholipase C, prevented myogenic responses of proximal but not distal ILA. Finally, basal proximal ILA diameters were increased by treatment with 50 nmol/L of staurosporine (P<0.05), and subsequent addition of thapsigargin (1 micromol/L) blocked myogenic contraction of proximal ILAs. Myogenic responses of intermediate ILAs exhibited characteristics between those of distal and proximal ILAs. Our data indicate that underlying mechanisms for myogenic responses differ in distinct segments of ILAs. The present results suggest that mechanosensitive cation channels are involved in myogenic constriction of distal ILAs. Finally, our findings provide evidence that the stimulation of phospholipase C mediates myogenic contraction of proximal ILAs. PMID- 9856975 TI - Chloride regulates afferent arteriolar contraction in response to depolarization. AB - -Renal vascular reactivity is influenced by the level of dietary salt intake. Recent in vitro data suggest that afferent arteriolar contractility is modulated by extracellular chloride. In the present study, we assessed the influence of chloride on K+-induced contraction in isolated perfused rabbit afferent arterioles. In 70% of vessels examined, K+-induced contraction was abolished by acute substitution of bath chloride. Consecutive addition of Cl- (30, 60, 80, 100, 110, and 117 mmol/L) restored the sensitivity to K+, and half-maximal response was observed at 82 mmol/L chloride. The calcium channel antagonist diltiazem (10(-6) mol/L) abolished K+-induced contractions. Bicarbonate did not modify the sensitivity to chloride. Norepinephrine (10(-6) mol/L) induced full contraction in depolarized vessels even in the absence of chloride. Iodide and nitrate were substituted for chloride with no inhibitory effect on K+-induced contraction. Approximately 30% of the vessels constricted in response to K+ in the absence of chloride. This response was reversibly blocked by the alpha1 blocker phentolamine (PA) (10(-5) mol/L) and, with PA present, the dependence on chloride was similar to the above series. The results show that K+-induced contraction of smooth muscle cells in the afferent arteriole is highly sensitive to chloride, whereas neurotransmitter release and ensuing contraction is not dependent on chloride. Thus, there are different activation pathways for depolarizing vasoconstrictors and for the sympathetic nervous system in renal afferent arterioles. This could be of physiological relevance for the resetting of afferent arteriolar sensitivity during changes in salt intake. PMID- 9856976 TI - Thiazide-induced vasodilation in humans is mediated by potassium channel activation. AB - -Hydrochlorothiazide and indapamide are thought to exert their hypotensive efficacy through a combined vasodilator and diuretic effect, but in vivo evidence for a direct vascular effect is lacking. The presence and mechanism of a direct vascular action of hydrochlorothiazide in vivo in humans were examined and compared with those of the thiazide-like drug indapamide. Forearm vasodilator responses to infusion of placebo and increasing doses of hydrochlorothiazide (8, 25, and 75 microg. min-1. dL-1) into the brachial artery were recorded by venous occlusion plethysmography. Dose-response curves were repeated after local tetraethylammonium (TEA) administration to determine the role of potassium channel activation and, in patients with the Gitelman syndrome, to determine the role of the thiazide-sensitive Na-Cl cotransporter in the vasodilator effect of hydrochlorothiazide. Vascular effects of hydrochlorothiazide were compared with those of indapamide in both normotensive (mean arterial pressure, 85+/-7 mm Hg) and hypertensive (mean arterial pressure, 124+/-16 mm Hg) subjects. At the highest infusion rate, local plasma concentrations of hydrochlorothiazide averaged 11.0+/-1.6 microg/mL, and those of indapamide averaged 7. 2+/-1.5 microg/mL. In contrast to indapamide, hydrochlorothiazide showed a direct vascular effect (maximal vasodilation, 55+/-14%; P=0. 013), which was inhibited by TEA (maximal vasodilation after TEA, 13+/-10%; P=0.02). The response was not dependent on blood pressure and was similar in patients with Gitelman syndrome, indicating that absence of the Na-Cl cotransporter does not alter the vasodilatory effect of hydrochlorothiazide. The vasodilator effect of hydrochlorothiazide in the human forearm is small and only occurs at high concentrations. The mechanism of action is not mediated by inhibition of vascular Na-Cl cotransport but involves vascular potassium channel activation. In contrast, indapamide does not exert any direct vasoactivity in the forearm vascular bed. PMID- 9856977 TI - Renal 11beta-hydroxysteroid dehydrogenase in genetically salt-sensitive hypertensive rats. AB - -Renal 11beta-hydroxysteroid dehydrogenase II (11beta-HSDII) converts glucocorticoids into inactive metabolites and plays an important role in controlling blood pressure and sodium retention. To examine whether this enzyme may be involved in the pathophysiology of salt-sensitive hypertension, we determined 11beta-HSDII activity and mRNA levels in the blood vessel and kidney of Dahl Iwai salt-sensitive (DS) rats and Dahl Iwai salt-resistant (DR) rats. Urinary free corticosterone:free 11-dehydrocorticosterone ratio was measured to estimate renal 11beta-HSD activity. Vascular 11beta-HSDII activity was expressed as percent conversion of [3H]corticosterone to [3H]11-dehydrocorticosterone in homogenized mesenteric arteries. 11beta-HSDII mRNA was estimated with the use of competitive polymerase chain reaction (PCR). Renal 11beta-HSDII activity and mRNA levels were significantly decreased in 8- and 12-week-old high salt DS rats compared with DR, Sprague-Dawley (SD), or low salt DS rats of the same age. Decreased 11beta-HSDII activity and mRNA levels in mesenteric arteries were observed in 8- and 12-week-old high salt DS rats. Urinary excretion of 11beta HSDII inhibitory factors was measured by inhibition of enzyme activity in microsomes from human kidney. The urinary inhibitors were significantly increased in 8- and 12-week-old high salt DS rats compared with DR, SD, or low salt DS rats of the same age. There were no significant differences in 11beta-HSDII activity and mRNA levels in mesenteric arteries and kidney or in urinary inhibitors between 4-week-old DS, DR, and SD rats. These results indicate that 11beta-HSDII may play a role in salt sensitivity and development of hypertension in the DS rat. PMID- 9856978 TI - Glucocorticoids inhibit superoxide anion production and p22 phox mRNA expression in human aortic smooth muscle cells. AB - -Recent reports suggest that the increased production of reactive oxygen species (ROS) in the vascular wall may contribute to the functional and structural changes associated with hypertension and atherosclerosis. Although glucocorticoid therapy can promote atherosclerosis, protective effects of these compounds on vascular lesion formation have been reported. In the present study, we investigated whether ROS production in cultured human aortic smooth muscle cells (HSMCs) can be modulated by glucocorticoids. Pretreatment of HSMCs with dexamethasone for 24 hours attenuated the basal and platelet-derived growth factor (PDGF)-AB- and angiotensin II-induced superoxide anion (O2. -) production. PDGF-AB-stimulated O2. - production was also inhibited by prednisolone and hydrocortisone but not by other steroids, such as testosterone and norgestrel. Incubation of HSMCs with glucocorticoids for 24 hours decreased 2',7' dichlorodihydrofluorescein (DCHF) oxidation, an indicator of intracellular ROS levels. Dexamethasone decreased the mRNA expression of p22 phox, one of the components of NADPH oxidase, but had no effect on the activity of superoxide dismutase. The effects of dexamethasone on DCHF oxidation, and p22 phox mRNA expression and PDGF-AB-stimulated O2. - production were inhibited by the glucocorticoid receptor antagonist RU486. These results indicate that glucocorticoids decrease O2. - production by HSMCs via a receptor-dependent pathway. This effect is likely to be mediated by a decrease in the generating system, such as downregulation of p22 phox mRNA, rather than an increased inactivation of O2. -. The inhibition of ROS production might contribute to the local protective effects that glucocorticoids have on vascular lesion formation. PMID- 9856979 TI - Arachidonate-induced tyrosine phosphorylation of epidermal growth factor receptor and Shc-Grb2-Sos association. AB - -Protein tyrosine phosphorylation induced by arachidonic acid (AA), an important lipid second messenger, was investigated in rabbit renal proximal tubule epithelial cells. AA stimulated tyrosine phosphorylation of a number of proteins with estimated molecular weights of 42, 44, 52, 56, 85, and 170/180 kDa. The phosphoproteins pp44 and pp42 were identified as 2 isoforms of mitogen-activated protein kinase (MAPK). Phosphorylation of MAPK in response to AA was transient, dose-dependent, and accompanied by an increase in its activity. The mechanism of AA-induced MAPK activation in RTE cells was protein kinase C-independent and involved tyrosine phosphorylation of adaptor protein Shc and its association with Grb2-Sos complex. Moreover, stimulation of RTE cells with AA resulted in significant phosphorylation of epidermal growth factor (EGF) receptor and its association with Shc. The effect of AA on EGF receptor phosphorylation, its association with Shc, and MAPK activation was similar to the effect of 1 ng/mL EGF. Tyrphostin AG1478, a specific inhibitor of EGF receptor tyrosine kinase activity, completely blocked the effects of AA and EGF but not phorbol ester on MAPK phosphorylation. These data suggest that in renal tubular epithelial cells, the mechanism of AA-induced MAPK activation involves tyrosine phosphorylation of EGF receptor and its association with Shc and Grb2-Sos complex. Given the critical role of AA in signaling linked to G protein-coupled receptors (GPCRs), these observations provide a mechanism for cross talk between GPCRs linked to phospholipases and the tyrosine kinase receptor signaling cascades. PMID- 9856980 TI - G-protein beta3 subunit gene (GNB3) variant in causation of essential hypertension. AB - -Essential hypertensives display enhanced signal transduction through pertussis toxin-sensitive G proteins. The T allele of a C825T variant in exon 10 of the G protein beta3 subunit gene (GNB3) induces formation of a splice variant (Gbeta3 s) with enhanced activity. The T allele of GNB3 was shown recently to be associated with hypertension in unselected German patients (frequency=0.31 versus 0.25 in control). To confirm and extend this finding in a different setting, we performed an association study in Australian white hypertensives. This involved an extensively examined cohort of 110 hypertensives, each of whom were the offspring of 2 hypertensive parents, and 189 normotensives whose parents were both normotensive beyond age 50 years. Genotyping was performed by polymerase chain reaction and digestion with BseDI, which either cut (C allele) or did not cut (T allele) the 268-bp polymerase chain reaction product. T allele frequency in the hypertensive group was 0.43 compared with 0.25 in the normotensive group (chi2=22; P=0.00002; odds ratio=2.3; 95% CI=1.7 to 3.3). The T allele tracked with higher pretreatment blood pressure: diastolic=105+/-7, 109+/-16, and 128+/ 28 mm Hg (mean+/-SD) for CC, CT, and TT, respectively (P=0.001 by 1-way ANOVA). Blood pressures were higher in female hypertensives with a T allele (P=0.006 for systolic and 0.0003 for diastolic by ANOVA) than they were in male hypertensives. In conclusion, the present study of a group with strong family history supports a role for a genetically determined, physiologically active splice variant of the G protein beta3 subunit gene in the causation of essential hypertension. PMID- 9856981 TI - Decreased flow-induced dilation and increased production of cGMP in spontaneously hypertensive rats. AB - -We investigated flow (shear stress)- and agonist-induced cGMP release in mesenteric vascular beds of spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). The mesenteric vascular bed was perfused in situ with Tyrode's solution. Vascular relaxation and cGMP release in the perfusate were determined on stimulation by flow or by acetylcholine (0.1 micromol/L) or sodium nitroprusside (0.1 mmol/L). Flow-induced release of cGMP was significantly greater in SHR than in WKY (P<0.01), despite a lower flow induced dilation in SHR. In both strains, NG-nitro-L-arginine methyl ester (L NAME) completely inhibited cGMP release in response to flow (P<0.001), although flow-induced dilation was not affected by L-NAME in SHR. Moreover, the activity of the constitutive nitric oxide synthase (NOS) was significantly greater in SHR (82+/-3.5 fmol/min) than in WKY (66+/-3.5 fmol/min; P<0.05) and was associated with increased expression of endothelial NOS mRNA in SHR. Sodium nitroprusside induced larger increases in cGMP release in SHR (3593+/-304 fmol/min) than in WKY (2467+/-302 fmol/min; P<0.05). The release of cGMP in response to acetylcholine was significantly lower in SHR (292+/-80 fmol/min) than in WKY (798+/-218 fmol/min; P<0.05) in parallel with smaller acetylcholine-induced relaxation in SHR. Despite increased cGMP production in response to flow and NOS activity, flow induced dilation was decreased in SHR, suggesting an upregulation of the NO/cGMP pathway to compensate for the increased vascular tone in SHR. PMID- 9856982 TI - Concordant genetic and physiological data are required for candidate genes. PMID- 9856983 TI - Phosphorylation of moesin by rho-associated kinase (Rho-kinase) plays a crucial role in the formation of microvilli-like structures. AB - Rho-associated kinase (Rho-kinase), which is activated by the small GTPase Rho, phosphorylates moesin at Thr558 in vitro. Here, using a site- and phosphorylation state-specific antibody, we found that the expression of dominant active RhoA in COS7 cells induced moesin phosphorylation and the formation of microvilli-like structures at apical membranes where the Thr558-phosphorylated moesin accumulated, whereas the expression of dominant negative Rho-kinase inhibited both of these processes. The expression of dominant active Rho-kinase also induced moesin phosphorylation. When COS7 cells expressing moesin or moesinT558A (substitution of Thr by Ala) were cultured under serum-depleted conditions, there were few microvilli-like structures, whereas microvilli-like structures remained in the cells expressing moesinT558D (substitution of Thr by Asp). The expression of moesinT558A inhibited the dominant active RhoA-induced formation of microvilli like structures. These results indicate that Rho-kinase regulates moesin phosphorylation downstream of Rho in vivo and that the phosphorylation of moesin by Rho-kinase plays a crucial role in the formation of microvilli-like structures. PMID- 9856984 TI - Assembly of the sarcoglycan complex. Insights for muscular dystrophy. AB - Four unique transmembrane glycoproteins comprise the sarcoglycan complex in striated muscle. The sarcoglycan complex contributes to maintenance of sarcolemma integrity. A shared feature of four types of autosomal recessive limb girdle muscular dystrophy (LGMD) is that mutations in a single sarcoglycan gene result in the loss of all sarcoglycans at the sarcolemma. The mechanism of destabilization is unknown. We report here our findings of sarcoglycan complex biosynthesis in a heterologous cell system. We demonstrate that the sarcoglycans are glycosylated and assemble into a complex that resides in the plasma membrane. Complex assembly was dependent on the simultaneous synthesis of all four sarcoglycans. Mutant sarcoglycans block complex formation and insertion of the sarcoglycans into the plasma membrane. This constitutes the first biochemical evidence to support the idea that the molecular defect in sarcoglycan-deficient LGMD is because of aberrant sarcoglycan complex assembly and trafficking, which leads to the absence of the complex from the sarcolemma. PMID- 9856985 TI - Identification of STRAP, a novel WD domain protein in transforming growth factor beta signaling. AB - Transforming growth factor-beta1 (TGF-beta1) is the prototype of a large family of proteins that regulate a variety of biological processes. The pleiotropic responses to TGF-beta are mediated via ligand-induced heteromeric complex formation by type I (TbetaR-I) and type II (TbetaR-II) serine-threonine kinase receptors. Several studies have shown that TbetaR-II acts as a primary receptor, binding TGF-beta and phosphorylating TbetaR-I, whose kinase activity then propagates the signals. Therefore, intracellular proteins that interact with type I receptors are likely to play important roles in TGF-beta signaling. We have identified a novel WD domain-containing protein, designated STRAP (serine threonine kinase receptor-associated protein), which interacts with TbetaR-I in a yeast two-hybrid system. STRAP associates with both functional TbetaR-I and TbetaR-II in vivo. Overexpression of STRAP leads to inhibition of TGF-beta mediated transcriptional activation. It also shows synergistic inhibition of TGF beta signaling in concert with Smad7, but not with Smad6, as measured by TGF-beta dependent transcriptional reporters. The existence of the STRAP gene from yeast to mammals indicates an evolutionarily conserved function in eukaryotes. The data suggest a potential role for STRAP in TGF-beta signal transduction. PMID- 9856986 TI - Expression of SFT (stimulator of Fe transport) is enhanced by iron chelation in HeLa cells and by hemochromatosis in liver. AB - SFT (stimulator of Fe transport) is a novel transport protein that has been found to facilitate uptake of iron presented to cells as either Fe(II) or Fe(III). When HeLa cells are exposed to the iron chelator desferrioxamine, levels of SFT mRNA increase in an actinomycin D-sensitive manner. In contrast, cells exposed to high levels of iron down-regulate SFT expression in a time-dependent and reversible fashion. Thus, homeostatic regulation of SFT expression not only ensures that sufficient levels of iron are maintained but also limits excessive assimilation to prevent potentially harmful effects of this toxic metal. The unexpected observation that SFT transcript levels are up-regulated in hemochromatosis patients therefore suggests that enhanced SFT expression contributes to the etiology of this iron overload disorder. PMID- 9856987 TI - Association of N- and C-terminal domains of phospholipase D is required for catalytic activity. AB - Rat brain phospholipase D1 (rPLD1) belongs to a superfamily defined by the highly conserved catalytic motif (H(X)K(X)4D, denoted HKD. RPLD1 contains two HKD domains, located in the N- and C-terminal regions. Deletion mutants of rPLD1 that contained only an N- or C-terminal HKD domain exhibited no catalytic activity when expressed in COS 7 cells. However, when N-terminal fragments containing one of the HKD domains were cotransfected with a C-terminal fragment containing the other HKD domain, PLD activity was restored. Furthermore, immunoprecipitation assays showed that the N- and C-terminal halves of rPLD1 were physically associated when expressed in COS 7 cells. In addition, deletion of 168 amino acids from the N terminus of rPLD1 significantly enhanced basal PLD activity while inhibiting the response to phorbol ester. Likewise, the coexpression of this truncated N-terminal half with the C-terminal half resulted in increased PLD activity. In summary, this study provides direct evidence that the enzymatic activity of rPLD1 requires the presence of the HKD domains in both the N- and C terminal regions of the molecule. More importantly, the two halves of rPLD1 can associate, and this may be essential to bring the two HKD domains together to form an active catalytic center. These findings provide new insights into the catalytic mechanism of enzymes of the PLD superfamily. PMID- 9856988 TI - Neurosecretion competence, an independently regulated trait of the neurosecretory cell phenotype. AB - Neurosecretion competence is intended as the ability of neurosecretory cells to express dense and clear vesicles discharged by regulated exocytosis (neurotransmitter release). Such a property, which so far has never been studied independently, is investigated here by a heterotypic cell fusion approach, using a clone of rat pheochromocytoma PC12 cells totally incompetent for neurosecretion that still largely maintains its typical molecular and cellular phenotype. When fused with wild-type partners of various species (rat, human) and specialization (PC12, neuroblastoma SH-SY5Y, HeLa), the defective cells reacquire their competence as revealed by the expression of their secretion-specific proteins. Fused wild-type cells therefore appear able to complement defective cells by providing them with factor(s) inducing the reactivation of their secretory program. The mechanism of action of these factors may consist not in a coordinate unblocking of transcription but in the prevention of a rapid post-transcriptional degradation of the mRNAs for secretion-specific genes. PMID- 9856989 TI - The N-terminal domain of RGS4 confers receptor-selective inhibition of G protein signaling. AB - Regulators of heterotrimeric G protein signaling (RGS) proteins are GTPase activating proteins (GAPs) that accelerate GTP hydrolysis by Gq and Gi alpha subunits, thus attenuating signaling. Mechanisms that provide more precise regulatory specificity have been elusive. We report here that an N-terminal domain of RGS4 discriminated among receptor signaling complexes coupled via Gq. Accordingly, deletion of the N-terminal domain of RGS4 eliminated receptor selectivity and reduced potency by 10(4)-fold. Receptor selectivity and potency of inhibition were partially restored when the RGS4 box was added together with an N-terminal peptide. In vitro reconstitution experiments also indicated that sequences flanking the RGS4 box were essential for high potency GAP activity. Thus, RGS4 regulates Gq class signaling by the combined action of two domains: 1) the RGS box accelerates GTP hydrolysis by Galphaq and 2) the N terminus conveys high affinity and receptor-selective inhibition. These activities are each required for receptor selectivity and high potency inhibition of receptor-coupled Gq signaling. PMID- 9856990 TI - Bile acid uptake via the human apical sodium-bile acid cotransporter is electrogenic. AB - Intestinal absorption of bile acids depends on a sodium-bile acid cotransport protein in the apical membrane of the ileal epithelial cell. Transport is Na+ dependent, but the Na+-bile acid stoichiometry and electrogenicity of transport are not known. Studies in whole intestine, isolated cells, and ileal membrane vesicles have been unable to resolve this issue because transport currents are small and can be obscured by other ionic conductances and transport proteins present in these membranes. In this study, the human apical sodium-bile acid transporter was expressed in stably transfected Chinese hamster ovary cells that lack other bile acid transporters. The Na+-dependent transport of a fluorescent bile acid analog, chenodeoxycholyl-Nepsilon-nitrobenzoxadiazol-lysine, was monitored by fluorescence microscopy in single, voltage-clamped cells. Bile acid movement was bidirectional and voltage-dependent with negative intracellular voltage-stimulating influx. A 3-fold reduction in extracellular Na+ produced a negative 52 mV shift of the flux-voltage relationship, consistent with a 2:1 Na+:bile acid coupling stoichiometry. No Na+- or voltage-dependent uptake was observed in nontransfected Chinese hamster ovary cells. These results indicate that the cotransport of bile acids and Na+ by human apical sodium-bile acid transporter is electrogenic and bidirectional and is best explained by a 2:1 Na+:bile acid coupling stoichiometry. These results suggest that membrane potential may regulate bile acid transport rates under physiological and pathophysiological conditions. PMID- 9856991 TI - Evidence for a functional interaction between integrins and G protein-activated inward rectifier K+ channels. AB - Heteromultimeric G protein-activated inward rectifier K+ (GIRK) channels, abundant in heart and brain, help to determine the cellular membrane potential as well as the frequency and duration of electrical impulses. The sequence arginine glycine-aspartate (RGD), located extracellularly between the first membrane spanning region and the pore, is conserved among all identified GIRK subunits but is not found in the extracellular domain of any other cloned K+ channels. Many integrins, which, like channels, are integral membrane proteins, recognize this RGD sequence on other proteins, usually in the extracellular matrix. We therefore asked whether GIRK activity might be regulated by direct interaction with integrin. Here, we present evidence that mutation of the RGD site to RGE, particularly on the GIRK4 subunit, decreases or abolishes GIRK current. Furthermore, wild-type channels can be co-immunoprecipitated with integrin. The total cellular amount of expressed mutant GIRK channel protein is the same as the wild-type protein; however, the amount of mutant channel protein that localizes to the plasma membrane is decreased relative to wild-type, most likely accounting for the diminished GIRK current detected. GIRK channels appear to bind directly to integrin and to require this interaction for proper GIRK channel membrane localization and function. PMID- 9856992 TI - Formation of alpha-Pal/Max heterodimers synergistically activates the eIF2-alpha promoter. AB - The transcription factor alpha-Pal recognizes two tandem palindromic repeats within the promoter of eukaryotic translation initiation factor 2-alpha (eIF2 alpha). Whereas both binding sites have the same "core domain" sequence (CGCATGCG), they differ with respect to their flanking sequences. Of the two sites, the 5'-cap proximal site has a higher binding affinity for alpha-Pal than does the 5'-cap distal site (Jacob, W. F., Silverman, T. A., Cohen, R. B., and Safer, B. (1989) J. Biol. Chem. 264, 20372-20384). The well characterized transcription factor Max binds to sequences that are remarkably similar to the core domain that alpha-Pal recognizes. To date, all of the Max heterodimer partners lack DNA binding domains and are thus dependent on Max interacting with DNA. Here we report that the two alpha-Pal sites have very different binding activities with respect to the E-box-binding protein Max. The 5'-cap distal or low alpha-Pal affinity site binds both alpha-Pal and Max. Furthermore, both heterodimers and homodimers of each of these proteins bind to this site. In contrast to the low affinity site, the high affinity site does not bind Max as a homodimer. This is the first documented case where Max heterodimerizes with a transcription factor that has affinity for DNA independent of Max. PMID- 9856993 TI - Two-stage activation for alpha5beta1 integrin binding to surface-adsorbed fibronectin. AB - By analyzing the functional binding of alpha5beta1 integrin to adsorbed fibronectin in intact cells, we demonstrate that integrin activation results in linear increases in adhesion strength as a function of ligand density, suggesting that modulation of the receptor-ligand interaction is the dominant mechanism for adhesion during the initial stages of adhesion and that cooperative binding contributes little to initial adhesion strength. Using this experimental framework, we show the existence of three distinct activation states for alpha5beta1 integrin binding to adsorbed fibronectin for both passive, antibody induced and active, cell-controlled activation. During the initial phase of adhesion, alpha5beta1 integrin is activated in an energy-dependent process from the nonbinding ground state to an intermediate state in which the receptor binds fibronectin and provides significant mechanical coupling. In later stages of adhesion maturation, alpha5beta1 integrin is activated to a higher binding state, which provides significant increases in adhesion strength compared with the intermediate state. These multiple binding states most likely result from different integrin conformations and reflect distinct interactions between alpha5beta1 and sites on adsorbed fibronectin. Multiple activation states for alpha5beta1 suggest the existence of distinct stages in adhesion signaling and strengthening and can provide a versatile mechanism for the regulation of adhesive interactions. PMID- 9856994 TI - Neurexophilin binding to alpha-neurexins. A single LNS domain functions as an independently folding ligand-binding unit. AB - alpha-Neurexins (Ialpha, IIalpha, and IIIalpha) are receptor-like proteins expressed in hundreds of isoforms on the neuronal cell surface. The extracellular domains of alpha-neurexins are composed of six LNS repeats, named after homologous sequences in the Laminin A G domain, Neurexins, and Sex hormone binding globulin, with three interspersed epidermal growth factor-like domains. Purification of neurexin Ialpha revealed that it is tightly complexed to a secreted glycoprotein called neurexophilin 1. Neurexophilin 1 is a member of a family of at least four genes and resembles a neuropeptide, suggesting a function as an endogenous ligand for alpha-neurexins. We have now used recombinant proteins and knockout mice to investigate which isoforms and domains of different neurexins and neurexophilins interact with each other. We show that neurexophilins 1 and 3 but not 4 (neurexophilin 2 is not expressed in rodents) bind to a single individual LNS domain, the second overall LNS domain in all three alpha-neurexins. Although this domain is alternatively spliced, all splice variants bind, suggesting that alternative splicing does not regulate binding. Using homologous recombination to disrupt the neurexophilin 1 gene, we generated mutant mice that do not express detectable neurexophilin 1 mRNA. Mice lacking neurexophilin 1 are viable with no obvious morbidity or mortality. However, homozygous mutant mice exhibit male sterility, probably because homologous recombination resulted in the co-insertion into the neurexophilin gene of herpes simplex virus thymidine kinase, which is known to cause male sterility. In the neurexophilin 1 knockout mice, neurexin Ialpha is complexed with neurexophilin 3 but not neurexophilin 4, suggesting that neurexophilin 1 is redundant with neurexophilin 3 and that neurexophilins 1 and 3 but not 4 bind to neurexins. This hypothesis was confirmed using expression experiments. Our data reveal that the six LNS and three epidermal growth factor domains of neurexins are independently folding ligand-binding domains that may interact with distinct targets. The results support the notion that neurexophilins represent a family of extracellular signaling molecules that interact with multiple receptors including all three alpha-neurexins. PMID- 9856995 TI - In situ flow activates endothelial nitric oxide synthase in luminal caveolae of endothelium with rapid caveolin dissociation and calmodulin association. AB - Acute changes in pressure or shear stress induce the rapid release of nitric oxide (NO) from the vascular endothelium resulting in vasodilation. Endothelial nitric oxide synthase (eNOS) regulates this flow-induced NO secretion. The subcellular location of flow-induced eNOS activity in the endothelium in vivo as well as the mechanisms by which hemodynamic forces regulate eNOS activity are unknown. The luminal cell surface of the endothelium, which is directly exposed to circulating blood stressors, has been examined for eNOS expression and functional activity. Immunoelectron microscopy of rat lung tissue shows eNOS labeling on the endothelial cell surface primarily within caveolae. Subcellular fractionation to purify luminal endothelial cell plasma membranes and their caveolae directly from rat lungs reveals that eNOS is not only concentrated but also enzymatically active in caveolae. Increasing vascular flow and pressure in situ rapidly activates caveolar eNOS with apparent eNOS dissociation from caveolin and association with calmodulin. Hemodynamic forces resulting from increased flow appear to transmit through caveolae to release eNOS from its inhibitory association with caveolin, apparently to allow more complete activation by calmodulin and other possible effectors. These data demonstrate a physiological relevant mechanotransduction event directly in caveolae at the luminal endothelial cell surface. Caveolae may serve as flow-sensing organelles with the necessary molecular machinery to transduce rapidly, mechanical stimuli and thereby regulate eNOS activity. PMID- 9856996 TI - Investigation of the anticoagulant mechanisms of a covalent antithrombin-heparin complex. AB - Recently, we developed a covalent antithrombin-heparin complex (ATH) as a possible treatment for respiratory distress syndrome. ATH reacted rapidly with thrombin and efficiently catalyzed the inhibition of either thrombin or factor Xa by exogenous antithrombin. In order to investigate mechanisms for the conjugate's unusual anticoagulant properties, changes in fluorescence due to covalent linkage or addition of exogenous antithrombin were studied in relation to reaction with thrombin derivatives or factor Xa. The emission spectrum of ATH was similar to that of antithrombin plus heparin mixtures. ATH quickly inhibited thrombin or factor Xa activities, as measured by a fluorogenic substrate. Fluorescein-labeled heparin was displaced from either thrombin or active site blocked thrombin by ATH, indicating that thrombin must bind to the conjugate's heparin moiety. Interaction of thrombin with ATH's heparin component was confirmed by a slow reaction rate of conjugate with a thrombin mutant that has weak heparin binding. Total intrinsic fluorescence increased when exogenous antithrombin was added to ATH, indicating that the catalytic mechanism may occur through a second inhibitor binding site. Thus, ATH reacts directly with thrombin through a bridge mechanism and probably catalyzes the reaction of thrombin with antithrombin by a second binding sequence on its heparin chain. PMID- 9856997 TI - Raf-1 is involved in the regulation of the interaction between guanine nucleotide exchange factor and Ha-ras. Evidences for a function of Raf-1 and phosphatidylinositol 3-kinase upstream to Ras. AB - The observation that activated c-Ha-Ras p21 interacts with diverse protein ligands suggests the existence of mechanisms that regulate multiple interactions with Ras. This work studies the influence of the Ras effector c-Raf-1 on the action of guanine nucleotide exchange factors (GEFs) on Ha-Ras in vitro. Purified GEFs (the catalytic domain of yeast Sdc25p and the full-length and catalytic domain of mouse CDC25Mm) and the Ras binding domains (RBDs) of Raf-1 (Raf (1-149) and Raf (51-131)) were used. Our results show that not only the intrinsic GTP/GTP exchange on Ha-Ras but also the GEF-stimulated exchange is inhibited in a concentration-dependent manner by the RBDs of Raf. Conversely, the scintillation proximity assay, which monitors the effect of GEF on the Ras.Raf complex, showed that the binding of Raf and GEF to Ha-Ras.GTP is mutually exclusive. The various GEFs used yielded comparable results. It is noteworthy that under more physiological conditions mimicking the cellular GDP/GTP ratio, Raf enhances the GEF-stimulated GDP/GTP exchange on Ha-Ras, in agreement with the sequestration of Ras.GTP by Raf. Consistent with our results, the GEF-stimulated exchange of Ha Ras.GTP was also inhibited by another effector of Ras, the RBD (amino acid residues 133-314) of phosphatidylinositol 3-kinase p110alpha. Our data show that Raf-1 and phosphatidylinositol 3-kinase can influence the upstream activation of Ha-Ras. The interference between Ras effectors and GEF could be a regulatory mechanism to promote the activity of Ha-Ras in the cell. PMID- 9856999 TI - A mutational approach shows similar mechanisms of recognition for the isolated and integrated versions of a protein epitope. AB - Antibody mAb164 is directed against the native form of the TrpB2 subunit of Escherichia coli tryptophan synthase. It recognizes a synthetic peptide, P11, constituted of residues 273-283 of TrpB, with high affinity. We introduced 16 single and 3 double mutations in each of the two contexts, TrpB2 and P11, and used them as local probes to study the cross-reactivity of mAb164 toward these two antigens. The equilibrium constant, KD, of dissociation from mAb164 was measured for each of the mutant derivatives of TrpB2 and P11 by a competition enzyme-linked immunosorbent assay and compared with the wild type one. The variation of the free energy of interaction, DeltaDeltaG, covered nearly 8 kcal/mol for the different mutations. The values of DeltaDeltaG for the mutant derivatives of TrpB2 and for those of P11 were close and the two sets of values were strongly correlated (r = 0.96). This correlation showed that mAb164 recognized the integrated and isolated versions of residues 273-283 with very similar mechanisms. A few significant differences between the recognitions of TrpB2 and P11 by mAb164 suggested some adaptability of the interaction. The results were compatible with a recognition of residues 273-283 of TrpB in a loop conformation, close to their structure in the crystals of the complete tryptophan synthase, TrpA2TrpB2. PMID- 9856998 TI - The propeptide domain of membrane type 1 matrix metalloproteinase is required for binding of tissue inhibitor of metalloproteinases and for activation of pro gelatinase A. AB - Activation of secreted latent matrix metalloproteinases (MMPs) is accompanied by cleavage of the N-terminal propeptide, thereby liberating the active zinc from binding to the conserved cysteine in the pro-domain. It has been assumed that an analogous mechanism is responsible for the activation of membrane type 1 MMP (MT1 MMP). Using recombinant wild-type MT1-MMP cDNA and mutant cDNAs transfected into COS-1 cells lacking endogenous MT1-MMP, we have examined the function of the propeptide domain of MT1-MMP. MT1-MMP was characterized by immunoblotting, surface biotinylation, gelatin substrate zymography, and 125I-tissue inhibitor of metalloproteinases 2 (TIMP-2) binding. In contrast to wild-type MT1-MMP transfected COS-1 cells, transfected COS-1 cells containing a deletion of the N terminal propeptide domain of MT1-MMP or a chimeric construction (substitution of the pro-domain of MT1-MMP with that of collagenase 3) were functionally inactive in terms of binding of 125I-labeled TIMP-2 to the cell surface and initiating the activation of pro-gelatinase A. These results support the concept that in its native plasma membrane-inserted form, the pro-domain of MT1-MMP plays an essential role in TIMP-2 binding and subsequent activation of pro-gelatinase A. PMID- 9857000 TI - Formation and fate of tyrosine. Intracellular partitioning of newly synthesized tyrosine in mammalian liver. AB - Tyrosine in an hepatocyte is transported from the plasma, synthesized from phenylalanine, or released during protein turnover. Effects of phenylalanine and tyrosine on the formation and fate (partitioning) of tyrosine from the different sources were examined in primary rat hepatocyte cultures. Rates of tyrosine degradation, transport, incorporation into and release from protein, and synthesis from phenylalanine were measured as well as the intracellular dilution of labeled tyrosine and phenylalanine incorporated into protein. We found tyrosine had little effect on phenylalanine hydroxylation over a wide range of conditions, that transported tyrosine and tyrosine from phenylalanine are in different metabolic pools, and that there appears to be channeling of newly synthesized tyrosine during degradation. In addition, under some conditions, intracellular partitioning of tyrosine is determined by tyrosine concentration. Specifically, if extracellular tyrosine is low and phenylalanine is at a normal plasma level, tyrosine use in protein synthesis takes precedence over tyrosine degradation or export. It is proposed that the mechanism controlling this is kinetic, based on relative rates of tyrosyl-tRNA formation and tyrosine degradation and export. A quantitative model of tyrosine and phenylalanine in flow and out-flow in hepatocytes is given, incorporating tyrosine synthesis, degradation, plasma membrane transport, and tyrosine and phenylalanine use and release during protein turnover. PMID- 9857001 TI - Characterization of c-myc 3' to 5' mRNA decay activities in an in vitro system. AB - The levels of mRNA and protein encoded by the c-myc protooncogene set the balance between proliferation and differentiation of mammalian cells. Thus, it is essential for the cell to tightly control c-myc expression. Indeed, cells utilize many mechanisms to control c-myc expression, including transcription, RNA processing, translation, and protein stability. We have focused on turnover of c myc mRNA as a key modulator of the timing and level of c-myc expression. c-myc mRNA is labile in cells, and its half-life is controlled by multiple instability elements located within both the coding region and the 3'-untranslated region (3' UTR). Much work has focused on the protein factors that bind the instability elements, yet little is known about the enzymatic activities that effect the degradation of c-myc mRNA. Here I have utilized a novel cell-free mRNA decay system to characterize the c-myc mRNA decay machinery. This machinery consists of 3' to 5' mRNA decay activities that are Mg2+-dependent, require neither exogenous ATP/GTP nor an ATP-regenerating system, and act independently of a 7mG(5')ppp(5')G cap structure to deadenylate an exogenous mRNA substrate in a c myc 3'-UTR-dependent fashion. Following deadenylation, nucleolytic decay of the 3'-UTR occurs generating 3' decay intermediates via a ribonucleolytic activity that can assemble on the c-myc 3'-UTR in a poly(A)-independent manner. PMID- 9857002 TI - The roles of nuclear factor of activated T cells and ying-yang 1 in activation induced expression of the interferon-gamma promoter in T cells. AB - Nuclear factor of activated T cells (NFAT) plays an important role in expression of many cytokine genes including interleukin-2 and interleukin-4. However, its role in interferon-gamma (IFN-gamma) expression is not well understood. In the current studies, two strong NFAT-binding sites in the IFN-gamma promoter were identified by DNase I footprint analysis at positions -280 to -270 and -163 to 155. NFATp bound independently to both sites and was required for the formation of a composite element with AP-1 spanning position -163 to -147. In Jurkat T cells and primary lymphocytes, activation-induced expression of IFN-gamma reporter constructs containing point mutations in either NFAT site or the AP-1 component of the composite site was decreased by approximately 40-65%. Despite elimination of both strong NFAT-binding sites, the IFN-gamma promoter remained completely sensitive to inhibition by cyclosporin. This suggests that other elements in the IFN-gamma promoter, such as the IFN-gamma proximal element, are sufficient for cyclosporin sensitivity of this gene. Ying-Yang 1 (YY1), a potential inhibitor of IFN-gamma expression, binds to sites located between the two NFAT sites. Mutation of the YY1 sites alone had little effect on IFN-gamma promoter activity. However, mutation of both the NFAT and YY1-binding sites abolished activation-induced expression in primary murine splenocytes but not in Jurkat T cells. This suggests that under some conditions, YY1 may play a positive role in activation-induced transcription of IFN-gamma. PMID- 9857003 TI - Insulin secretagogues activate the secretory granule receptor-like protein tyrosine phosphatase IAR. AB - To investigate the potential role of protein-tyrosine phosphatases (PTPs) in regulated secretion, cellular PTP activity was measured in pancreatic beta cell lines after exposure to insulin secretagogues. A peak of elevated PTP activity was detected in whole cell lysates after 15-20 min of treatment of the cells with high KCl, glucose, or TPA, which did not appear upon treatment with control compounds. Neither was it detected in cells that do not undergo regulated secretion. The PTP activation was transient, SDS-resistant, and localized to the cytoskeleton fraction of cells. The cytoskeletal localization of IAR, a receptor like PTP associated with secretory granules of neuroendocrine cells, suggested the possibility that IAR is the secretagogue-activated PTP. The transient expression of human IAR in betaTC3 and HIT-T15 beta cells, followed by treatment with secretagogues or control compounds and immunoprecipitation of human IAR, showed that immunoprecipitates from the secretagogue-treated cells contained an elevated PTP activity. The secretagogue-induced activation of IAR had identical kinetics to that of the endogenous PTP. Although ectopic IAR was present in membrane and cytoskeletal fractions from the cells, only the cytoskeleton associated IAR could be activated. Thus IAR represents the endogenous secretagogue-responsive PTP, or at least a component of it, and is one of the few receptor-like PTPs for which enzymatic activation has been demonstrated. Insulin secretion is detected prior to IAR activation, suggesting that IAR is not required for immediate secretion but likely plays a role in events downstream of insulin secretion or in another pathway related to the specialized function of secretory cells. PMID- 9857004 TI - Involvement of topoisomerases in the initiation of simian virus 40 minichromosome replication. AB - Topoisomerases provide the unlinking activity necessary for replication fork movement during DNA replication. It is uncertain, however, whether topoisomerases are also required for the initiation of replication. To investigate this point, we have performed pulse-chase experiments with SV40 minichromosomes as template to distinguish between the initiation and the elongation of replication. Using an unfractionated cytosolic extract as a source of replication functions, we found that the addition of topoisomerases at the initiation step significantly increased the number of active chromatin templates, whereas addition of topoisomerases at the elongation step had only minor effects. Minichromosomes with an extended chromatin structure as well as protein-free DNA required less topoisomerase for effective replication initiation. We could exclude the possibility that topoisomerases enhance the origin binding of T antigen, the SV40 replication initiator, and propose instead that the arrangement of nucleosomes influences the diffusion of supercoils during initial DNA unwinding. Efficient initiation therefore requires a high local concentration of topoisomerases to relax the torsional stress. PMID- 9857005 TI - The C331A mutant of neuronal nitric-oxide synthase is defective in arginine binding. AB - It has been proposed that Cys99 of human endothelial nitric oxide synthase (eNOS) is responsible for tetrahydrobiopterin (BH4) binding. To examine this possibility rigorously, we expressed rat neuronal NOS (nNOS) in Escherichia coli, with the homologous Cys331 to Ala mutation, and characterized structural and functional attributes of the purified, mutated enzyme. C331A-nNOS, as isolated, was catalytically incompetent. Upon prolonged incubation with L-arginine (L-Arg), not only BH4 binding but also catalytic activity could be restored. In contrast to wild-type nNOS (WT-nNOS), which exhibits an absorbance maximum at 407 nm that shifts immediately upon L-arginine addition to a high spin form, the C331A-nNOS mutant, as isolated, exhibited an absorbance maximum at 420 nm. C331A-nNOS, as isolated, did not bind detectable levels of either [3H]Nomega-nitro-L-arginine or [3H]BH4, but [3H]BH4 binding was reinstated after extended incubation with excess L-arginine. On the other hand, C331A-nNOS and WT-NOS were identical with regard to imidazole binding affinity, CaM binding affinity, and rates of cytochrome c and 2, 6-dichlorophenolindophenol reduction. EPR spectroscopy revealed conversion of low to high spin heme after extended incubation with high concentrations of L arginine (0.1-10 mM). The estimated Kd for L-arginine binding to C331A-nNOS was two orders of magnitude greater than WT-nNOS (>100 microM versus 2-3 microM). Here we propose that Cys331 plays an important role in stabilizing L-arginine binding to nNOS. Our findings suggest that the primary dysfunction in the C331A mutant of nNOS, as isolated, is disruption of the BH4-substrate binding interactions as broadcast from this mutated cysteine residue. Prolonged incubation with L-arginine appears to cause remodeling of the mutant protein to a form similar to that of WT-nNOS, allowing for normalized BH4 binding and nitric oxide synthetic activity. PMID- 9857006 TI - Structure-function relationships in a type I antifreeze polypeptide. The role of threonine methyl and hydroxyl groups in antifreeze activity. AB - Several analogs of an alanine-rich, alpha-helical type I antifreeze polypeptide from the winter flounder were synthesized and studied to evaluate the role of threonine residues on antifreeze activity. In this series, the four Thr residues in the wild type polypeptide were substituted with from one to four Ser, allo Thr, or Val residues. Circular dichroism studies determined that these substitutions did not significantly diminish alpha-helicity. Thermal hysteresis data showed that substitution of Thr by Ser resulted in moderate to complete loss of antifreeze activity, depending on the number and position of the substituted Thr residue(s). Replacement by Val, in confirmation of other recent reports, or by allo-Thr had a much less detrimental effect on activity though there were qualitative differences in activity between the mutants and the wild type AFP. Based on these results, we propose that both the methyl and hydroxyl groups of Thr, particularly of the central two Thr residues, Thr13 and Thr24, play key roles in the ice-binding properties of the antifreeze peptide. Specifically, the methyls participate in hydrophobic interactions with ice, which provide the driving force for binding and stability, whereas the hydroxyls and other polar residues control binding specificity and impart additional stability through hydrogen bonding. PMID- 9857007 TI - FK-binding protein is associated with the ryanodine receptor of skeletal muscle in vertebrate animals. AB - The ryanodine receptor/calcium release channel (RyR1) of sarcoplasmic reticulum from rabbit skeletal muscle terminal cisternae (TC) contains four tightly associated FK506-binding proteins (FKBP12). Dissociation and reconstitution studies have shown that RyR1 can be modulated by FKBP12, which helps to maintain the channel in the quiescent state. In this study, we found that the association of FKBP with RyR1 of skeletal muscle is common to each of the five classes of vertebrates. TC from skeletal muscle representing animals from different vertebrates, i.e. mammals (rabbit), birds (chicken), reptiles (turtle), fish (salmon and rainbow trout), and amphibians (frog), were isolated. For each, we find the following: 1) FKBP12 is localized to the TC (there are four FKBP binding sites/ryanodine receptor); 2) soluble FKBP exchanges with the bound form on RyR1 of TC; 3) release of FKBP from terminal cisternae by drug (FK590) treatment leads to a significant reduction in the net calcium loading rate, consistent with channel activation (the calcium loading rate is restored to the control value by reconstitution with FKBP12); and 4) RyR1 of skeletal muscle TC can bind to and exchange with either FKBP12 or FKBP12.6 (FKBP12.6 is the novel FKBP isoform found selectively associated with RyR2 of dog cardiac sarcoplasmic reticulum). We conclude that FKBP is an integral part of the RyR1 of skeletal muscle in each of the classes of vertebrate animals. The studies are consistent with a role for FKBP in skeletal muscle excitation-contraction coupling. PMID- 9857008 TI - Isolation and characterization of NUC70, a cytoplasmic, hematopoietic apoptotic endonuclease. AB - Endonucleolytic DNA fragmentation is the common end point and the prevailing indicator of apoptosis. We have identified a 70-kDa endonuclease (NUC70) that is activated in drug-induced apoptosis of human hematopoietic cells. We purified NUC70 to homogeneity and generated a rabbit polyclonal antibody to distinguish it from previously identified nucleases. Biochemical characterization of isolated NUC70 demonstrates that it is Ca2+/Mg2+-dependent and active over a pH range of 6 8. When incubated with isolated HeLa nuclei, NUC70 was capable of generating internucleosomal DNA fragmentation. This endonucleolytic activity was inhibited by Zn2+, aurintricarboxylic acid, N-ethylmaleimide, spermine, and iodoacetamide. Western immunoblots using the anti-NUC70 antibody and DNA-SDS-polyacrylamide gel electrophoresis assays indicate that NUC70 expression and activity is restricted to human hematopoietic cells. No such activity was detected in human epithelial cell lines or murine hematopoietic cells. We also observed no difference in levels of NUC70 expression between apoptotic and nonapoptotic cells, suggesting that activation of NUC70 may be by posttranslational modification. We demonstrate that NUC70 activity is diminished in cells pretreated with the caspase inhibitors z-DEVD-fmk, z-VAD-fmk, and Z-CH2-Asp-DCB. Time course studies of cytoplasmic and nuclear endonuclease activities during apoptosis show that NUC70 is a cytoplasmic endonuclease that is translocated to the nucleus after the initiation of apoptosis. We confirmed this with immunostaining studies using anti-NUC70 antibody. These results demonstrate that NUC70 is an endogenous cytoplasmic endonuclease that is activated during apoptosis in a caspase-dependent mechanism. PMID- 9857009 TI - Mammalian mitochondrial ribosomal proteins. N-terminal amino acid sequencing, characterization, and identification of corresponding gene sequences. AB - The integrity of healthy mitochondria is supposed to depend largely on proper mitochondrial protein biosynthesis. Mitochondrial ribosomal proteins (MRPs) are directly involved in this process. To identify mammalian mitochondrial ribosomal proteins and their corresponding genes, we purified mature rat MRPs and determined 12 different N-terminal amino acid sequences. Using this peptide information, data banks were screened for corresponding DNA sequences to identify the genes or to establish consensus cDNAs and to characterize the deduced MRP open reading frames. Eight different groups of corresponding mammalian MRPs constituted from human, mouse, and rat origin were identified. Five of them show significant sequence similarities to bacterial and/or yeast mitochondrial ribosomal proteins. However, MRPs are much less conserved in respect to the amino acid sequence among species than cytoplasmic ribosomal proteins of eukaryotes and bacteria. PMID- 9857010 TI - Single point mutations distributed in 10 soluble and membrane regions of the Nicotiana plumbaginifolia plasma membrane PMA2 H+-ATPase activate the enzyme and modify the structure of the C-terminal region. AB - The Nicotiana plumbaginifolia pma2 (plasma membrane H+-ATPase) gene is capable of functionally replacing the H+-ATPase genes of the yeast Saccharomyces cerevisiae, provided that the external pH is kept above 5.0. Single point mutations within the pma2 gene were previously identified that improved H+-ATPase activity and allowed yeast growth at pH 4.0. The aim of the present study was to identify most of the PMA2 positions, the mutation of which would lead to improved growth and to determine whether all these mutations result in similar enzymatic and structural modifications. We selected additional mutants in total 42 distinct point mutations localized in 30 codons. They were distributed in 10 soluble and membrane regions of the enzyme. Most mutant PMA2 H+-ATPases were characterized by a higher specific activity, lower inhibition by ADP, and lower stimulation by lysophosphatidylcholine than wild-type PMA2. The mutants thus seem to be constitutively activated. Partial tryptic digestion and immunodetection showed that the PMA2 mutants had a conformational change making the C-terminal region more accessible. These data therefore support the hypothesis that point mutations in various H+-ATPase parts displace the inhibitory C-terminal region, resulting in enzyme activation. The high density of mutations within the first half of the C-terminal region suggests that this part is involved in the interaction between the inhibitory C-terminal region and the rest of the enzyme. PMID- 9857011 TI - Synthesis of poly-N-acetyllactosamine in core 2 branched O-glycans. The requirement of novel beta-1,4-galactosyltransferase IV and beta-1,3-n acetylglucosaminyltransferase. AB - Poly-N-acetyllactosamine is a unique carbohydrate composed of N-acetyllactosamine repeats and provides the backbone structure for additional modifications such as sialyl Lex. Poly-N-acetyllactosamines in mucin-type O-glycans can be formed in core 2 branched oligosaccharides, which are synthesized by core 2 beta-1,6-N acetylglucosaminyltransferase. Using a beta-1, 4-galactosyltransferase (beta4Gal TI) present in milk and the recently cloned beta-1,3-N acetylglucosaminyltransferase, the formation of poly-N-acetyllactosamine was found to be extremely inefficient starting from a core 2 branched oligosaccharide, GlcNAcbeta1-->6(Galbeta1-->3)GalNAcalpha-->R. Since the majority of synthesized oligosaccharides contained N-acetylglucosamine at the nonreducing ends, galactosylation was judged to be inefficient, prompting us to test novel members of the beta4Gal-T gene family for this synthesis. Using various synthetic acceptors and recombinant beta4Gal-Ts, beta4Gal-TIV was found to be most efficient in the addition of a single galactose residue to GlcNAcbeta1- >6(Galbeta1-->3)GalNAcalpha-->R. Moreover, beta4Gal-TIV, together with beta-1,3-N acetylglucosaminyltransferase, was capable of synthesizing poly-N acetyllactosamine in core 2 branched oligosaccharides. On the other hand, beta4Gal-TI was found to be most efficient for poly-N-acetyllactosamine synthesis in N-glycans. In contrast to beta4Gal-TI, the efficiency of beta4Gal-TIV decreased dramatically as the acceptors contained more N-acetyllactosamine repeats, consistent with the fact that core 2 branched O-glycans contain fewer and shorter poly-N-acetyllactosamines than N-glycans in many cells. These results, as a whole, indicate that beta4Gal-TIV is responsible for poly-N acetyllactosamine synthesis in core 2 branched O-glycans. PMID- 9857012 TI - Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. AB - Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. Chem. 272, 27589-27597). In human liver extracts, most of the glycogenin-2 was only detectable after treatment with alpha-amylase. Similarly, purifed high Mr glycogen was only detected after release by alpha amylase treatment. Based on analysis by polymerase chain reaction, the predominant isoform in liver was glycogenin-2beta. Glycogenin-2 was found in Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate. Both human liver and human RD-ES cell extracts also contained glycogenin-1. Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. Mutation of Tyr-196 in glycogenin-2 to a Phe residue abolished the ability of glycogenin-2 to self-glucosylate but not to interact with glycogenin-1. Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5 fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. This result is important since it indicates that the level of glycogenin-2 can determine glycogen accumulation and hence has the potential to control glycogen synthesis. PMID- 9857013 TI - Single tottering mutations responsible for the neuropathic phenotype of the P type calcium channel. AB - Recent genetic and molecular biological analyses have revealed many forms of inherited channelopathies. Homozygous ataxic mice, tottering (tg) and leaner (tgla) mice, have mutations in the P/Q-type Ca2+ channel alpha1A subunit gene. Although their clinical phenotypes, histological changes, and locations of gene mutations are known, it remains unclear what phenotypes the mutant Ca2+ channels manifest, or whether the altered channel properties are the primary consequence of the mutations. To address these questions, we have characterized the electrophysiological properties of Ca2+ channels in cerebellar Purkinje cells, where the P-type is the dominant Ca2+ channel, dissociated from the normal, tg, and tgla mice, and compared them with the properties of the wild-type and mutant alpha1A channels recombinantly expressed with the alpha2 and beta subunits in baby hamster kidney cells. The most striking feature of Ca2+ channel currents of mutant Purkinje cells was a marked reduction in current density, being reduced to approximately 60 and approximately 40% of control in tg and tgla mice, respectively, without changes of cell size. The Ca2+ channel currents in the tg Purkinje cells showed a relative increase in non-inactivating component in voltage-dependent inactivation. Besides the same change, those of the tgla mice showed a more distinct change in voltage dependence of activation and inactivation, being shifted in the depolarizing direction by approximately 10 mV, with a broader voltage dependence of inactivation. In the recombinant expression system, the tg channel with a missense mutation (P601L) and one form of the two possible tgla aberrant splicing products, tgla (short) channel, showed a significant reduction in current density, while the other form of the tgla channels, tgla (long), had a current density comparable to the normal control. On the other hand, the shift in voltage dependence of activation and inactivation was observed only for the tgla (long) channel. Comparison of properties of the native and recombinant mutant channels suggests that single tottering mutations are directly responsible for the neuropathic phenotypes of reduction in current density and deviations in gating behavior, which lead to neuronal death and cerebellar atrophy. PMID- 9857014 TI - The sec-independent twin-arginine translocation system can transport both tightly folded and malfolded proteins across the thylakoid membrane. AB - A subset of lumen proteins is transported across the thylakoid membrane by a Sec independent translocase that recognizes a twin-arginine motif in the targeting signal. A related system operates in bacteria, apparently for the export of redox cofactor-containing proteins. In this report we describe a key feature of this system, the ability to transport folded proteins. The thylakoidal system is able to transport dihydrofolate reductase (DHFR) when an appropriate signal is attached, and the transport efficiency is almost undiminished by the binding of folate analogs such as methotrexate that cause the protein to fold very tightly. The system is moreover able to transport DHFR into the lumen with methotrexate bound in the active site, demonstrating that the DeltapH-driven transport of large, native structures is possible by this pathway. However, correct folding is not a prerequisite for transport. Truncated, malfolded DHFR can be translocated by this system, as can physiological substrates that are severely malfolded by the incorporation of amino acid analogs. PMID- 9857015 TI - Recombinant glutathione S-transferase/CD36 fusion proteins define an oxidized low density lipoprotein-binding domain. AB - CD36 is a multifunctional cell-surface receptor that binds adhesion molecules such as thrombospondin-1 and collagen and modified lipids and/or lipoproteins. It participates in cellular uptake of photoreceptor outer segments and scavenging of apoptotic cells and oxidized low density lipoprotein (Ox-LDL). Recognition and internalization of Ox-LDL by mononuclear phagocytes may play an important role in the development of atherosclerotic lesions. We have utilized a series of recombinant bacterial glutathione S-transferase/CD36 fusion proteins that span nearly all of the CD36 molecule to characterize the structural domain on CD36 that recognizes Ox-LDL. We found that the Ox-LDL-binding domain is different from the thrombospondin-1-binding domain located at amino acids 93-120. A fusion protein containing the region extending from amino acids 5 to 143 formed specific, saturable, and reversible complexes with Ox-LDL. As with intact CD36, binding was blocked by excess unlabeled Ox-LDL and antibodies to CD36. The stoichiometry and affinity of the fusion protein for Ox-LDL were similar to those of the intact protein. We also demonstrated that this fusion protein competitively inhibited binding of Ox-LDL to purified platelet CD36 and to CD36 expressed on peripheral blood monocytes and CD36 cDNA-transfected melanoma cells. The use of smaller peptides and fusion proteins including those spanning amino acids 28-93 and 5-93 has further narrowed the binding site to a region from amino acids 28 to 93, although participation of a sequence in the noncontiguous region 120-155 cannot be excluded. This study, for the first time, demonstrates unique regions of the scavenger receptor CD36 that bind the Ox-LDL ligand. Our structural analysis of the receptor provides information as to potential control of the trafficking of modified lipoproteins into the blood vessel wall. PMID- 9857016 TI - Linoleic acid-induced activity of plant uncoupling mitochondrial protein in purified tomato fruit mitochondria during resting, phosphorylating, and progressively uncoupled respiration. AB - An uncoupling protein was recently discovered in plant mitochondria and demonstrated to function similarly to the uncoupling protein of brown adipose tissue. In this work, green tomato fruit mitochondria were purified on a self generating Percoll gradient in the presence of 0.5% bovine serum albumin to deplete mitochondria of endogenous free fatty acids. The uncoupling protein activity was induced by the addition of linoleic acid during the resting state, and in the progressively uncoupled state, as well as during phosphorylating respiration in the presence of benzohydroxamic acid, an inhibitor of the alternative oxidase and with succinate (+ rotenone) as oxidizable substrate. Linoleic acid strongly stimulated the resting respiration in fatty acid-depleted mitochondria but had no effect on phosphorylating respiration, suggesting no activity of the uncoupling protein in this respiratory state. Progressive uncoupling of state 4 respiration decreased the stimulation by linoleic acid. The similar respiratory rates in phosphorylating and fully uncoupled respiration in the presence and absence of linoleic acid suggested that a rate-limiting step on the dehydrogenase side of the respiratory chain was responsible for the insensitivity of phosphorylating respiration to linoleic acid. Indeed, the ADP/O ratio determined by ADP/O pulse method was decreased by linoleic acid, indicating that uncoupling protein was active during phosphorylating respiration and was able to divert energy from oxidative phosphorylation. Moreover, the respiration rates appeared to be determined by membrane potential independently of the presence of linoleic acid, indicating that linoleic acid-induced stimulation of respiration is due to a pure protonophoric activity without any direct effect on the electron transport chain. PMID- 9857017 TI - Molecular basis for the stabilization and inhibition of 2, 3-dihydroxybiphenyl 1,2-dioxygenase by t-butanol. AB - The steady-state cleavage of catechols by 2,3-dihydroxybiphenyl 1, 2-dioxygenase (DHBD), the extradiol dioxygenase of the biphenyl biodegradation pathway, was investigated using a highly active, anaerobically purified preparation of enzyme. The kinetic data obtained using 2,3-dihydroxybiphenyl (DHB) fit a compulsory order ternary complex mechanism in which substrate inhibition occurs. The Km for dioxygen was 1280 +/- 70 microM, which is at least 2 orders of magnitude higher than that reported for catechol 2,3-dioxygenases. Km and Kd for DHB were 22 +/- 2 and 8 +/- 1 microM, respectively. DHBD was subject to reversible substrate inhibition and mechanism-based inactivation. In air-saturated buffer, the partition ratios of catecholic substrates substituted at C-3 were inversely related to their apparent specificity constants. Small organic molecules that stabilized DHBD most effectively also inhibited the cleavage reaction most strongly. The steady-state kinetic data and crystallographic results suggest that the stabilization and inhibition are due to specific interactions between the organic molecule and the active site of the enzyme. t-Butanol stabilized the enzyme and inhibited the cleavage of DHB in a mixed fashion, consistent with the distinct binding sites occupied by t-butanol in the crystal structures of the substrate-free form of the enzyme and the enzyme-DHB complex. In contrast, crystal structures of complexes with catechol and 3-methylcatechol revealed relationships between the binding of these smaller substrates and t-butanol that are consistent with the observed competitive inhibition. PMID- 9857018 TI - Characterization of the Escherichia coli damage-independent UvrBC endonuclease activity. AB - Incision of damaged DNA templates by UvrBC in Escherichia coli depends on UvrA, which loads UvrB on the site of the damage. A 50-base pair 3' prenicked DNA substrate containing a cholesterol lesion is incised by UvrABC at two positions 5' to the lesion, the first incision at the eighth and the second at the 15th phosphodiester bond. Analysis of a 5' prenicked cholesterol substrate revealed that the second 5' incision is efficiently produced by UvrBC independent of UvrA. This UvrBC incision was also found on the same substrate without a lesion and, with an even higher efficiency, on a DNA substrate containing a 5' single strand overhang. Incision occurred in the presence of ATP or ADP but not in the absence of cofactor. We could show an interaction between UvrB and UvrC in solution and subsequent binding of this complex to the substrate with a 5' single strand overhang. Analysis of mutant UvrB and UvrC proteins revealed that the damage independent nuclease activity requires the protein-protein interaction domains, which are exclusively needed for the 3' incision on damaged substrates. However, the UvrBC incision uses the catalytic site in UvrC which makes the 5' incision on damaged DNA substrates. PMID- 9857019 TI - Myocyte enhancer factor 2C and Nkx2-5 up-regulate each other's expression and initiate cardiomyogenesis in P19 cells. AB - The Nkx2-5 homeodomain protein plays a key role in cardiomyogenesis. Ectopic expression in frog and zebrafish embryos results in an enlarged myocardium; however, expression of Nkx2-5 in fibroblasts was not able to trigger the development of beating cardiac muscle. In order to examine the ability of Nkx2-5 to modulate endogenous cardiac specific gene expression in cells undergoing early stages of differentiation, P19 cell lines overexpressing Nkx2-5 were differentiated in the absence of Me2SO. Nkx2-5 expression induced cardiomyogenesis in these cultures aggregated without Me2SO. During differentiation into cardiac muscle, Nkx2-5 expression resulted in the activation of myocyte enhancer factor 2C (MEF2C), but not MEF2A, -B, or -D. In order to compare the abilities of Nkx2-5 and MEF2C to induce cellular differentiation, P19 cells overexpressing MEF2C were aggregated in the absence of Me2SO. Similar to Nkx2-5, MEF2C expression initiated cardiomyogenesis, resulting in the up regulation of Brachyury T, bone morphogenetic protein-4, Nkx2-5, GATA-4, cardiac alpha-actin, and myosin heavy chain expression. These findings indicate the presence of a positive regulatory network between Nkx2-5 and MEF2C and show that both factors can direct early stages of cell differentiation into a cardiomyogenic pathway. PMID- 9857020 TI - Mutations that induce constitutive activation and mutations that impair signal transduction modulate the basal and/or agonist-stimulated internalization of the Lutropin/Choriogonadotropin receptor. AB - Previous results from this laboratory suggested that the same active conformation of the lutropin/choriogonadotropin receptor (LHR) is involved in the stimulation of G proteins and in triggering the internalization of the bound agonist. We have now analyzed two naturally occurring, constitutively active mutants of the human LHR. These mutations were introduced into the rat LHR (rLHR) and are designated L435R and D556Y. Cells expressing rLHR-D556Y bind human choriogonadotropin (hCG) with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation does not affect the internalization of the free receptor, but it enhances the internalization of the agonist-occupied receptors approximately 3-fold. Cells expressing rLHR-L435R also bind hCG with normal affinity, exhibit a 47-fold increase in basal cAMP, and do not respond to hCG with a further increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors approximately 2- and approximately 17-fold, respectively. We conclude that the state of activation of the rLHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing rLHR-L435R is due to the fast rate of internalization of the bound hCG. The finding that membranes expressing rLHR-L435R (a system where internalization does not occur) respond to hCG with an increase in adenylyl cyclase activity supports this suggestion. PMID- 9857021 TI - Biochemical characterization of tomato annexin p35. Independence of calcium binding and phosphatase activities. AB - Tomato annexin p35 has been cloned and used in a site-directed mutagenesis study to explore the phospholipid binding and catalytic properties of the protein in detail. Analysis of the cDNA sequence of p35 reveals that the annexin has only two typical endonexin folds, corresponding to repeats I and IV. Expression of recombinant p35 in Escherichia coli confirmed both phospholipid binding and a nucleotide phosphatase activity that could be inhibited on interaction of the recombinant annexin with phospholipids. Site-directed mutagenesis in which the acidic residues Glu-68 (repeat I), and Asp-297 (repeat IV) were changed to Asn, generated two mutant forms, E68N and D297N, respectively. Both mutant forms of the annexin continued to express catalytic activity. Changing repeat I had little effect on phospholipid binding, whereas the change to repeat IV abolished this property. These data show that, in this plant annexin, repeat IV plays a more critical role in calcium-dependent phospholipid binding than repeat I, and that the catalytic and phospholipid binding activity of the protein can be separated experimentally. PMID- 9857022 TI - Mouse mu opioid receptor gene expression. A 34-base pair cis-acting element inhibits transcription of the mu opioid receptor gene from the distal promoter. AB - The 5'-flanking region of the mouse mu opioid receptor (MOR) gene has two promoters, referred to as distal and proximal, and the activities of each in the brain are quite different from each other. The 5'-distal promoter regulatory sequences (5'-DPRS), positioned between these two promoters, have strong inhibitory effects on the reporter gene expression driven by the MOR distal promoter. In our studies, detailed 3' deletion mapping of the 5'-DPRS narrowed down the negative cis-acting element to a 34-base pair (bp) segment (position 721 to -687). This 34-bp cis-acting element functions in both neuronal (NMB) and non-neuronal (CHO and RAW264.7) cultured cells. S1 nuclease protection assays indicated that this 34-bp cis-acting element suppresses distal promoter activity at the transcriptional level. Linker scanning mutagenesis demonstrated that nucleotides around position -721 and -689 in the 34-bp cis-acting element are essential for the regulation of distal promoter activity. Operational characterization of the 34-bp cis-acting element in the homologous MOR distal promoter and the heterologous SV40 promoter showed that its effects are position- and promoter-dependent while being orientation-independent in both promoters. Collectively, these data suggested that this 34-bp segment is a conditional transcriptional cis-acting element that blocks mouse MOR gene expression from the distal promoter. PMID- 9857024 TI - Catalytic mechanism and specificity for hydrolysis and transglycosylation reactions of cytosolic beta-glucosidase from guinea pig liver. AB - Cytosolic beta-glucosidase (CBG) from mammalian liver is known for its broad substrate specificity and has been implicated in the transformation of xenobiotic glycosides. CBG also catalyzes a variety of transglycosylation reactions, which have been been shown with other glycosylhydrolases to function in synthetic and genetic regulatory pathways. We investigated the catalytic mechanism, substrate specificity, and transglycosylation acceptor specificity of guinea pig liver CBG by several methods. These studies indicate that CBG employs a two-step catalytic mechanism with the formation of a covalent enzyme-sugar intermediate and that CBG will transfer sugar residues to primary hydroxyls and equatorial but not axial C 4 hydroxyls of aldopyranosyl sugars. Kinetic studies revealed that correction for transglycosylation reactions is necessary to derive correct kinetic parameters for CBG. Further analyses revealed that for aldopyranosyl substrates, the activation energy barrier is affected most by the presence of a C-6 carbon and by the configuration of the C-2 hydroxyl, whereas the binding energy is affected modestly by the configuration and substituents at C-2, C-4, and C-5. These data indicate that the transglycosylation activity of CBG derives from the formation of a covalently linked enzyme-sugar intermediate and that the specificity of CBG for transglycosylation reactions is different from its specificity for hydrolysis reactions. PMID- 9857023 TI - A TrkA-selective, fast internalizing nerve growth factor-antibody complex induces trophic but not neuritogenic signals. AB - Nerve growth factor (NGF) is a neurotrophin that induces neuritogenic and trophic signals by binding to TrkA and/or p75 receptors. We report a comparative study of the binding, internalization, and biological activity of NGF versus that of NGF in association with an anti-NGF monoclonal antibody (mAb NGF30), directed against the C termini of NGF. NGF.mAb complexes do not bind p75 effectively but bind TrkA with high affinity. After binding, NGF. mAb complexes stimulate internalization faster and to a larger degree than NGF. NGF.mAb-induced activation of TrkA, Shc, and MAPK is transient compared with NGF-induced activation; yet NGF and NGF. mAb afford identical trophic responses. In contrast, NGF induces Suc-1-associated neurotrophic activating protein phosphorylation and neuritogenic differentiation, but NGF.mAb does not. Thus, an absolute separation of trophic and neuritogenic function is seen for NGF.mAb, suggesting that biological response modifiers of neurotrophins can afford ligands with selected activities. PMID- 9857025 TI - Fatty acid synthase dimers containing catalytically active beta-ketoacyl synthase or malonyl/acetyltransferase domains in only one subunit can support fatty acid synthesis at the acyl carrier protein domains of both subunits. AB - A double-tagging, dual affinity chromatographic procedure, which permits isolation of dimers independently mutated in each subunit, has been exploited to probe the functional topology of the animal fatty acid synthase. Dimers were engineered in which the chain-terminating thioesterase reaction was compromised by mutation of the (active-site) serine residue in both subunits; these dimers assembled two long-chain fatty acyl moieties, which remained covalently linked to the 4'-phosphopantetheine residues of the two acyl carrier protein domains. Significantly, dimers that contained an additional mutation that compromised the activity of either the beta-ketoacyl synthase or malonyl/acetyltransferase activity in only one subunit also assembled two long-chain acyl moieties. In contrast, in a control experiment, introduction of an additional mutation that compromised the function of the acyl carrier protein domain in only one subunit resulted in the assembly of only one long-chain acyl moiety per dimer. Because the beta-ketoacyl synthase and malonyl/acetyltransferase domains are located near the amino terminus of the polypeptide and the acyl carrier protein domain near the carboxyl terminus, these results support a modified model for the animal fatty acid synthase in which head-to-tail functional contacts are possible both within as well as between subunits. PMID- 9857026 TI - Cloning and characterization of GEF-H1, a microtubule-associated guanine nucleotide exchange factor for Rac and Rho GTPases. AB - The Rho-related small GTPases are critical elements involved in regulation of signal transduction cascades from extracellular stimuli to cell nucleus and cytoskeleton. The Dbl-like guanine nucleotide exchange factors (GEF) have been implicated in direct activation of these GTPases. Here we have identified a new member of the Dbl family, GEF-H1, by screening a human HeLa cell cDNA library. GEF-H1 encodes a 100-kDa protein containing the conserved structural array of a Dbl homology domain in tandem with a pleckstrin homology domain and is most closely related to the lfc oncogene, but additionally it contains a unique coiled coil domain at the carboxyl terminus. Biochemical analysis reveals that GEF-H1 is capable of stimulating guanine nucleotide exchange of Rac and Rho but is inactive toward Cdc42, TC10, or Ras. Moreover, GEF-H1 binds to Rac and Rho proteins in both the GDP- and guanosine 5'-3-O-(thio)triphosphate-bound states without detectable affinity for Cdc42 or Ras. Immunofluorescence reveals that GEF-H1 colocalizes with microtubules through the carboxyl-terminal coiled-coil domain. Overexpression of GEF-H1 in COS-7 cells results in induction of membrane ruffles. These results suggest that GEF-H1 may have a direct role in activation of Rac and/or Rho and in bringing the activated GTPase to specific target sites such as microtubules. PMID- 9857027 TI - Simultaneous measurements of cytosolic and mitochondrial Ca2+ transients in HT29 cells. AB - Loading of HT29 cells with the Ca2+ dye fura-2/AM resulted in an nonhomogeneous intracellular distribution of the dye. Cellular compartments with high fura-2 concentrations were identified by correlation with mitochondrial markers, cellular autofluorescence induced by UV, and dynamic measurement of autofluorescence after inhibition of oxidative phosphorylation. Stimulation with carbachol (10(-4) mol/liter) increased cytosolic, nuclear, and mitochondrial Ca2+ activity ([Ca2+]c, [Ca2+]n, and [Ca2+]m, respectively) measured by UV confocal and conventional imaging. Similar results were obtained with a prototype two photon microscope (Zeiss, Jena, Germany) allowing for fura-2 excitation. The increase of [Ca2+]m lagged behind that of [Ca2+]c and [Ca2+]n by 10-20 s, and after removing the agonist, [Ca2+]m also decreased with a delay. A strong increase of [Ca2+]m occurred only when a certain threshold of [Ca2+]c (around 1 micromol/liter) was exceeded. In a very similar way, ATP, neurotensin, and thapsigargin increased [Ca2+]c and [Ca2+]m. Carbonyl cyanide p trifluoromethoxyphenylhyrdrazone reversibly reduced the increase of [Ca2+]m. The source of the mitochondrial Ca2+ increase had intra- and extracellular components, as revealed by experiments in low extracellular Ca2+. We conclude that agonist-induced Ca2+ signals are transduced into mitochondria. 1) Mitochondria could serve as a Ca2+ sink, 2) mitochondria could allow the modulation of [Ca2+]c and [Ca2+]n signals, and 3) [Ca2+]m may serve as a stimulatory metabolic signal when a cell is highly stimulated. PMID- 9857028 TI - Generation of destabilized green fluorescent protein as a transcription reporter. AB - The green fluorescent protein (GFP) is a widely used reporter in gene expression and protein localization studies. GFP is a stable protein; this property allows its accumulation and easy detection in cells. However, this stability also limits its application in studies that require rapid reporter turnover. We created a destabilized GFP for use in such studies by fusing amino acids 422-461 of the degradation domain of mouse ornithine decarboxylase (MODC) to the C-terminal end of an enhanced variant of GFP (EGFP). The fusion protein, unlike EGFP, was unstable in the presence of cycloheximide and had a fluorescence half-life of 2 h. Western blot analysis indicated that the fluorescence decay of EGFP-MODC-(422 461) was correlated with degradation of the fusion protein. We mutated key amino acids in the PEST sequence of EGFP-MODC-(422-461) and identified several mutants with variable half-lives. The suitability of destabilized EGFP as a transcription reporter was tested by linking it to NFkappaB binding sequences and monitoring tumor necrosis factor alpha-mediated NFkappaB activation. We obtained time course induction and dose response kinetics similar to secreted alkaline phosphatase obtained in transfected cells. This result did not occur when unmodified EGFP was used as the reporter. Because of its autofluorescence, destabilized EGFP can be used to directly correlate gene induction with biochemical change, such as NFkappaB translocation to the nucleus. PMID- 9857029 TI - The rub family of ubiquitin-like proteins. Crystal structure of Arabidopsis rub1 and expression of multiple rubs in Arabidopsis. AB - Several proteins with significant identity to ubiquitin have been characterized recently. In contrast to ubiquitin's main role in targeting proteins for degradation, a described function of one family of ubiquitin-related proteins, the Rub family, is to serve as a stable post-translational modification of a complex involved in the G1-to-S cell cycle transition. Rub proteins have been found in animals, plants, and fungi and consist of 76 residues with 52-63% identity to ubiquitin. In this study three different RUB proteins within the plant Arabidopsis are identified; two differ by only 1 amino acid, while the third is only 77.6% identical to the other two. Genes encoding all three are expressed in multiple organs. In addition, we report the crystal structure of higher plant RUB1 at 1.7-A resolution to help elucidate the functional differences between Rub and ubiquitin. RUB1 contains a single globular domain with a flexible COOH-terminal extension. The overall RUB1 structure is very similar to ubiquitin. The majority of the amino acid differences between RUB1 and ubiquitin map to the surface. These changes alter the electrostatic surface potential in two regions and likely confer specificity between ubiquitin and RUB1 and their ubiquitin-activating enzyme (E1) or E1-like activating enzymes. PMID- 9857030 TI - Crystal structure of the human ubiquitin-like protein NEDD8 and interactions with ubiquitin pathway enzymes. AB - The NEDD8/Rub1 class of ubiquitin-like proteins has been implicated in progression of the cell cycle from G1 into S phase. These molecules undergo a metabolism that parallels that of ubiquitin and involves specific interactions with many different proteins. We report here the crystal structure of recombinant human NEDD8 refined at 1.6-A resolution to an R factor of 21.9%. As expected from the high sequence similarity (57% identical), the NEDD8 structure closely resembles that reported previously for ubiquitin. We also show that recombinant human NEDD8 protein is activated, albeit inefficiently, by the ubiquitin activating (E1) enzyme and that NEDD8 can be transferred from E1 to the ubiquitin conjugating enzyme E2-25K. E2-25K adds NEDD8 to a polyubiquitin chain with an efficiency similar to that of ubiquitin. A chimeric tetramer composed of three ubiquitins and one histidine-tagged NEDD8 binds to the 26 S proteasome with an affinity similar to that of tetraubiquitin. Seven residues that differ from the corresponding residues in ubiquitin, but are conserved between NEDD8 orthologs, are candidates for mediating interactions with NEDD8-specific partners. One such residue, Ala-72 (Arg in ubiquitin), is shown to perform a key role in selecting against reaction with the ubiquitin E1 enzyme, thereby acting to prevent the inappropriate diversion of NEDD8 into ubiquitin-specific pathways. PMID- 9857031 TI - Structure-activity of tetrad-forming oligonucleotides as a potent anti-HIV therapeutic drug. AB - Recently, we have described the design and characterization of oligonucleotides containing only G and T bases, i.e. T30695 and T30177, that are potent inhibitors of human immunodeficiency virus type 1 (HIV-1) replication in culture (Jing, N., Rando, R. F., Pommier, Y., and Hogan, M. E. (1997) Biochemistry 36, 12498-12505). To understand that observation and to rationalize the generally high thermal stability of oligonucleotide folding for these compounds, we have used NMR methods, coupled to molecular modeling, to obtain a high resolution structure model for T30695, which is the most potent of the integrase inhibitors that have been identified thus far. Modeling and NMR data obtained in the presence of Li+ ions show that T30695 assumes an intramolecular fold with a distorted G-octet core and a set of three open, partially disordered loops. This is referred to as Li+-form structure. The NMR-based model suggests that, upon coordination with three K+ equivalents, the central G-octet becomes more regular and that the loop domains become orderly and compact. This is referred to as K+-form structure. Based upon the assay of inhibition of HIV-1 integrase, T30695 demonstrated a strong inhibition of HIV-1 integrase activity as the K+-form structure, but a poor inhibition of HIV-1 integrase activity as the Li+-form structure. The structure/activity analysis suggests that the K+-induced conformation transition of the tetrad-forming oligonucleotides, such as T30695 and T30177, plays a key role in inhibition of HIV-1 integrase activity. PMID- 9857032 TI - Endocytosis of functional epidermal growth factor receptor-green fluorescent protein chimera. AB - A chimera of the epidermal growth factor receptor (EGFR) and green fluorescent protein (GFP) has been engineered by fusing GFP to the carboxyl terminus of EGFR. Data are provided to demonstrate that the GFP moiety does not affect the expected functioning of EGFR. EGFR-GFP becomes phosphorylated at tyrosine residues in response to EGF and is capable of phosphorylating endogenous substrates and initiating signaling cascades. EGF-dependent association of the chimeric receptor with the clathrin adaptor protein AP-2, involved in endocytosis, and with Shc adaptor protein, which binds in close proximity to the fusion point, is not affected by the GFP moiety. Receptor down-regulation and internalization occur at rates similar to those in cells expressing wild-type EGFR. Western blot analysis reveals that lysosomal degradation of EGFR-GFP proceeds from the extracellular domain and that GFP is not preferentially cleaved. Time-dependent co-localization of EGFR-GFP and Texas Red-conjugated EGF in living cells using digital deconvolution microscopy demonstrates the trafficking of ligand-receptor complexes through the early and multivesicular endosomes followed by segregation of the ligand and receptor at the late stages of endocytosis. Time-lapse optical analysis of the early stages of endocytosis reveals localization of EGFR-GFP in the tubular-vesicular endosomal compartments. Rapid dynamics of membrane movement and fusion within these compartments were observed. This approach and the fidelity of the biochemical properties of the EGFR-GFP demonstrate that real-time visualization of trafficking and protein interactions of tyrosine kinase receptors in the presence or absence of the ligand are feasible. PMID- 9857033 TI - Molecular cloning and characterization of a novel retinoic acid-inducible gene that encodes a putative G protein-coupled receptor. AB - The effects of retinoids such as all-trans-retinoic acid (ATRA) on cell growth, differentiation, and apoptosis are thought to be mediated by nuclear retinoid receptors, which are involved in ligand-dependent transcriptional activation of target genes. Using differential display, we identified the cDNA of a novel gene, designated retinoic acid-inducible gene 1 (RAIG1), which was induced by ATRA in the squamous carcinoma cell line UMSCC-22B. Two RAIG1 transcripts of 2.4 and 6.8 kilobase pairs, respectively, have the same ORF that encodes a 357-amino acid polypeptide. RAIG1 mRNA is expressed at high level in fetal and adult lung tissues. Induction of RAIG1 expression by ATRA is rapid (within 2 h) and dose dependent in the range between 1 nM to 1 microM. The constitutive RAIG1 mRNA levels, which were low in three of five head and neck and four of six lung cancer cell lines, increased after ATRA treatment in most cell lines. The deduced RAIG1 protein sequence contains seven transmembrane domains, characteristic of G protein-coupled receptors. A fusion protein of RAIG1 and the green fluorescent protein was localized in the cell surface membrane and perinuclear vesicles in transiently transfected cells. RAIG1 was mapped to chromosome 12p12. 3-p13. Our results provide novel evidence for a possible interaction between retinoid and G protein signaling pathways. PMID- 9857034 TI - Electron microscopy and x-ray diffraction studies of Lotus tetragonolobus A isolectin cross-linked with a divalent Lewisx oligosaccharide, an oncofetal antigen. AB - The interactions of lectins with multivalent carbohydrates often leads to the formation of highly ordered cross-linked lattices that are amenable to structural studies. A particularly well ordered, two-dimensional lattice is formed from fucose-specific isolectin A from Lotus tetragonolobus cross-linked with difucosyllacto-N-neohexaose, an oligosaccharide possessing the Lewisx determinant, which is an oncofetal antigen. A combination of electron microscopy, x-ray diffraction, simulation of electron micrographs, and molecular model building was used to determine the relative positions of the tetrameric lectin and bivalent carbohydrate within the lattice. X-ray diffraction from unoriented pellets was used to determine the lattice dimensions and analysis of electron micrographs was used to determine the lattice symmetry. Molecular models of the lattice were constructed based on the known structure of the jack bean lectin concanavalin A and the high degree of sequence homology between the two lectins. Using the symmetry and dimensions of the lattice and its appearance in filtered electron micrographs, molecular models were used to determine the orientation of the lectin in the lattice, and to define the range of lectin-oligosaccharide interactions consistent with the structural data. The present study provides the first description of a highly ordered, two-dimensional, cross-linked lattice between a tetravalent lectin and a bivalent carbohydrate. PMID- 9857035 TI - Accurate 3' end processing and adenylation of human signal recognition particle RNA and alu RNA in vitro. AB - Human signal recognition particle (SRP) RNA is transcribed by RNA polymerase III and terminates with -GUCUCUUUUOH on its 3' end. Our previous studies showed that the three terminal uridylic acid residues of human SRP RNA are post transcriptionally removed and a single adenylic acid residue is added, resulting in a 3' end sequence of -GUCUCUAOH (Sinha, K. M., Gu, J., Chen, Y., and Reddy, R. (1998) J. Biol. Chem. 273, 6853-6859). In this study we show that the Alu RNA, corresponding to the 5' and 3' ends of SRP RNA, is also accurately processed and adenylated in vitro. Alu RNAs containing 7 or 11 additional nucleotides on the 3' end were accurately processed and then adenylated. Deletion analysis showed that an 87-nucleotide-long motif comprising of the 5' and 3' ends, including stem IV of the Alu RNA, is sufficient and necessary for the 3' end processing and adenylation. A 73-nucleotide-long construct with deletion of stem IV, required for the binding of SRP 9/14-kDa proteins, was neither processed nor adenylated. The adenylated Alu RNA as well as adenylated SRP RNA were bound to the SRP 9/14 kDa heterodimer and were immunoprecipitated by specific antibodies. A significant fraction of SRP RNA in the nucleoli was found to be processed and adenylated. These data are consistent with nascent SRP and/or Alu RNAs first binding to SRP 9/14-kDa protein heterodimer, followed by the removal of extra sequence on the 3' end and then the addition of one adenylic acid residue in the nucleus, before transport into the cytoplasm. PMID- 9857036 TI - The N-terminal sequence affects distant helix interactions in hemoglobin. Implications for mutant proteins from studies on recombinant hemoglobin felix. AB - The N-terminal 18-amino acid sequence of the beta-chain of hemoglobin, as far as the end of the A helix, has been replaced by the corresponding sequence of the gamma-chain of fetal hemoglobin with the remaining sequence of the beta-chain retained (helices B through H). The gamma-beta-chain had the correct mass, and its entire sequence was established by mass spectrometric analysis of its tryptic peptides; the alpha-chain also had the correct mass. This recombinant hemoglobin (named Hb Felix) retains cooperativity and has an oxygen affinity like that of HbA both in the presence and absence of the allosteric regulators, 2,3 diphosphoglycerate or chloride but differs from HbF in its 2,3-diphosphoglycerate response. However, Hb Felix has some features that resemble fetal hemoglobin, i.e. its significantly decreased tetramer-dimer dissociation and its circular dichroism spectrum, which measure the strength of the tetramer-dimer interface in the oxy conformation and its rearrangement to the deoxy conformation, respectively. Even though Hb Felix contains the HbA amino acids at its tetramer dimer interface, which is located at a distance from the substitution sites, its interface properties resemble those of HbF. Therefore, the N-terminal sequence and not just those amino acids directly involved at the subunit interface contacts with alpha-chains must have a strong influence on this region of the molecule. The results reinforce the concept of fluid long range relationships among various parts of the hemoglobin tetramer (Dumoulin, A., Manning, L. R., Jenkins, W. T., Winslow, R. M., and Manning, J. M. (1997) J. Biol. Chem. 272, 31326-31332) and demonstrate the importance of the N-terminal sequence, especially in some mutant hemoglobins, in influencing its overall structure by affecting the relationship between helices. PMID- 9857037 TI - Identification of an interaction between the m-band protein skelemin and beta integrin subunits. Colocalization of a skelemin-like protein with beta1- and beta3-integrins in non-muscle cells. AB - Signaling across integrins is regulated by interaction of these receptors with cytoskeletal proteins and signaling molecules. To identify molecules interacting with the cytoplasmic domain of the beta3-integrin subunit (glycoprotein IIIa), a placental cDNA library was screened in the yeast two-hybrid system. Two identical clones coding for a 96-amino acid sequence were identified. This sequence was 100% identical to a sequence in skelemin, a protein identified previously in skeletal muscle. Skelemin is a member of a superfamily of cytoskeletal proteins that contain fibronectin-type III-like motifs and immunoglobulin C2-like motifs and that regulate the organization of myosin filaments in muscle. The amino acid residues in the isolated clones encompassed C2 motifs 4 and 5 of skelemin. A recombinant skelemin protein consisting of C2 motifs 3-7 interacted with beta1- and beta3-integrin cytoplasmic domains expressed as glutathione S-transferase (GST) fusion proteins, but not with GST-beta2-integrin cytoplasmic tail or GST alone. The skelemin-binding region was in the membrane proximal cytoplasmic domains of the integrins. Full-length skelemin interacted with integrin in intact cells as demonstrated by the colocalization of hemagglutinin-tagged skelemin in Chinese hamster ovary (CHO) cells containing alphaIIbbeta3-integrin and by the finding that microinjection of C2 motif 4 of skelemin into C2C12 mouse myoblast cells caused spread cells to round up. A skelemin-like protein was detected in CHO cells, endothelial cells, and platelets, and this protein colocalized with beta1- and beta3-integrins in CHO cells. This study suggests the presence of a skelemin-like protein in non-muscle cells and provides evidence that it may be involved in linking integrins to the cytoskeleton. PMID- 9857038 TI - Dimerization/docking domain of the type Ialpha regulatory subunit of cAMP dependent protein kinase. Requirements for dimerization and docking are distinct but overlapping. AB - Based on increasing evidence that the type I R subunits as well as the type II R subunits localize to specific subcellular sites, we have carried out an extensive characterization of the stable dimerization domain at the N terminus of RIalpha. Deletion mutants as well as alanine scanning mutagenesis were used to delineate critical regions as well as particular amino acids that are required for homodimerization. A set of nested deletion mutants defined a minimum core required for dimerization. Two single site mutations on the C37H template, RIalpha(F47A) and RIalpha(F52A), were sufficient to abolish dimerization. In addition to serving as a dimerization motif, this domain also serves as a docking surface for binding to dual specificity anchoring proteins (D-AKAPs) (Huang, L. J., Durick, K., Weiner, J. A., Chun, J., and Taylor, S. S. (1997) J. Biol. Chem. 272, 8057-8064; Huang, L. J., Durick, K., Weiner, J. A., Chun, J., and Taylor, S. S. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 11184-11189). A similar strategy was used to map the sequence requirements for anchoring of RIalpha to D-AKAP1. Although dimerization appears to be essential for anchoring to D-AKAP1, anchoring can also be abolished by the following single site mutations: C37H, V20A, and I25A. These sites define "hot spots" for the anchoring surface since each of these dimeric proteins are deficient in binding to D-AKAP1. In contrast to earlier predictions, the alignment of the dimerization/docking domains of RIalpha and RII show striking similarities yet subtle differences not only in their secondary structure (Newlon, M. G., Roy, M., Hausken, Z. E., Scott, J. D., and Jennings. P. A. (1997) J. Biol. Chem. 272, 23637-23644) but also in the distribution of residues important for both docking and dimerization functions. PMID- 9857039 TI - The suppressor of cytokine signaling (SOCS) 1 and SOCS3 but not SOCS2 proteins inhibit interferon-mediated antiviral and antiproliferative activities. AB - The suppressor of cytokine signaling (SOCS) proteins are a family of cytokine inducible negative regulators of cytokine signaling. Interferon (IFN)-gamma treatment induces the expression of SOCS1, SOCS2, and SOCS3 mRNAs. To examine the effect of SOCS proteins on IFN-mediated Janus-activated kinase/signal transducers and activators of transcription (STAT) signaling, HeLa- and MCF-7-derived stable cell lines expressing SOCS1, SOCS2, and SOCS3 proteins were established. SOCS1 and SOCS3 but not SOCS2 inhibited the tyrosine phosphorylation and nuclear translocation of STAT1 in response to IFN stimulation. The IFN-mediated antiviral and antiproliferative activities were consistently blocked by the constitutive expression of SOCS1 and SOCS3 but not SOCS2 proteins. The maximum inhibitory activities of SOCS1 and SOCS3 proteins toward the activation of STAT1 were observed at very low levels of SOCS protein expression. In addition, SOCS1 exhibited a much stronger inhibitory activity toward the activation of STAT1 than did SOCS3. These results suggest that SOCS1 and SOCS3 but not SOCS2 are inhibitors of IFN-mediated Janus-activated kinase/STAT signaling pathways. PMID- 9857040 TI - Substrate specificity of the SpCCE1 holliday junction resolvase of Schizosaccharomyces pombe. AB - SpCCE1 from Schizosaccharomyces pombe is an endonuclease that resolves Holliday junctions in vitro. SpCCE1 also binds and cleaves a range of other DNAs (Y junction; flap; and flayed, nicked, and partial duplexes) with varying efficiency. Cleavage sites are always 3' of thymine nucleotides positioned at or close to the branch point or strand interruption. SpCCE1's favored substrate is the X-junction. Up to two dimers of SpCCE1 can bind concurrently to the same X junction at its crossover point. From mixing experiments of SpCCE1 and the Escherichia coli RuvA protein, we show that each dimer of SpCCE1 binds to a different face of the X-junction and that both are seemingly competent for strand cleavage. We propose that this provides a mechanism whereby SpCCE1 can scrutinize all four junction strands simultaneously for cleavable thymine nucleotides. SpCCE1 appears to resolve X-junctions by a nick and counter-nick mechanism. Therefore, to ensure a high probability of bilateral strand cleavage, SpCCE1 has a relatively long lifetime on X-junctions. This mechanism has the drawback of limiting dissociation from noncleavable junctions. We discuss why this might not be a problem in vivo. PMID- 9857041 TI - Early steps in the unfolding of thermolysin-like proteases. AB - Several series of site-directed mutations in thermolysin-like proteases are presented that show remarkable nonadditivity in their effect on thermal stability. A simple model is proposed that relates this nonadditivity to the occurrence of independent partial unfolding processes that occur in parallel at elevated temperatures. To prove this model, a thermolysin-like protease was designed in which two mutations located approximately 35 A apart in the structure individually exert small stabilizing effects of 2.3 and 4. 1 degreesC, respectively, but when combined stabilize the protease by 14.6 degreesC. This overadditivity, which follows directly from the model, confirms that unfolding of this engineered protease starts in parallel at two different regions of the protein. PMID- 9857042 TI - Structural determinants of metal-induced conformational changes in HIV-1 integrase. AB - Human immunodeficiency virus type 1 (HIV-1) integrase (IN) undergoes a reversible metal-induced conformational change that activates the enzyme (Asante-Appiah, E., and Skalka, A. M. (1997) J. Biol. Chem. 272, 16196-16205). In this report, key structural features that mediate this conformational change have been identified by site-directed mutagenesis, limited proteolysis, and mass spectrometry studies. The results reveal two separable metal-induced effects. One depends on residues in the N-terminal domain (amino acids 1-50) and a C-terminal tail (amino acids 274-288) and is detected by increased resistance of the full-length protein to proteolytic digestion. This effect appears to depend on metal binding at an undefined location distinct from the known sites in the N-terminal and catalytic core domains. The second conformational change depends on metal binding at the active site in the catalytic core domain. Substitution of acidic residues Asp64 or Glu152 in the catalytic core D,D(35)E motif or truncation of the Src homology 3 (SH3)-like domain in the C-terminal region of the enzyme abolishes this metal induced change. Comparison of tryptic digests of an HIV-1 IN derivative competent for metal-induced conformational change and a conformation-defective D64N derivative identified specific regions in HIV-1 IN that are affected by this second change. A region in the N terminus that spans Lys14, an extended loop and the adjacent region in the core domain (including lysines 136, 156, and 160 and Arg173), and residues at the C terminus beyond the SH3-like domain all become less accessible to proteolysis in the conformation-competent protein. In contrast, a region that encompasses Lys258 in the putative DNA binding groove of the SH3-like domain becomes more sensitive to proteolysis in the presence of Mn2+. The results are consistent with a model in which the binding of the metal ion by residues of the D,D(35)E motif elicits specific changes in all three domains of HIV-1 IN, inducing the restructuring of the enzyme for catalytic competence. PMID- 9857043 TI - Hyaluronan fragments synergize with interferon-gamma to induce the C-X-C chemokines mig and interferon-inducible protein-10 in mouse macrophages. AB - Hallmarks of chronic inflammation and tissue fibrosis are increased influx of activated inflammatory cells, mediator release, and increased turnover and production of the extracellular matrix (ECM). Recent evidence has suggested that fragments of the ECM component hyaluronan play a role in chronic inflammation by inducing macrophage expression of chemokines. Interferon-gamma (IFN-gamma), an important regulator of macrophage functions, has been shown to induce the C-X-C chemokines Mig and IP-10. These chemokines affect T-cell recruitment and inhibit angiogenesis. The purpose of this investigation was to determine the effect of hyaluronan (HA) on IFN-gamma-induced Mig and IP-10 expression in mouse macrophages. We found a marked synergy between HA and IFN-gamma on Mig and IP-10 mRNA and protein expression in mouse macrophages. This was most significant with Mig, which was not induced by HA alone. The synergy was specific for HA, was not dependent on new protein synthesis, was not mediated by tumor necrosis factor alpha, was selective for Mig and IP-10, and occurred at the level of gene transcription. These data suggest that the ECM component HA may influence chronic inflammatory states by working in concert with IFN-gamma to alter macrophage chemokine expression. PMID- 9857044 TI - Coexpression of the KCNA3B gene product with Kv1.5 leads to a novel A-type potassium channel. AB - Shaker-related voltage-gated potassium (Kv) channels may be heterooligomers consisting of membrane-integral alpha-subunits associated with auxiliary cytoplasmic beta-subunits. In this study we have cloned the human Kvbeta3.1 subunit and the corresponding KCNA3B gene. Identification of sequence-tagged sites in the gene mapped KCNA3B to band p13.1 of human chromosome 17. Comparison of the KCNA1B, KCNA2B, and KCNA3B gene structures showed that the three Kvbeta genes have very disparate lengths varying from >/=350 kb (KCNA1B) to approximately 7 kb (KCNA3B). Yet, the exon patterns of the three genes, which code for the seven known mammalian Kvbeta subunits, are very similar. The Kvbeta1 and Kvbeta2 splice variants are generated by alternative use of 5'-exons. Mouse Kvbeta4, a potential splice variant of Kvbeta3, is a read-through product where the open reading frame starts within the sequence intervening between Kvbeta3 exons 7 and 8. The human KCNA3B sequence does not contain a mouse Kvbeta4-like open reading frame. Human Kvbeta3 mRNA is specifically expressed in the brain, where it is predominantly detected in the cerebellum. The heterologous coexpression of human Kv1.5 and Kvbeta3.1 subunits in Chinese hamster ovary cells yielded a novel Kv channel mediating very fast inactivating (A-type) outward currents upon depolarization. Thus, the expression of Kvbeta3.1 subunits potentially extends the possibilities to express diverse A-type Kv channels in the human brain. PMID- 9857045 TI - Chlorophyll a formation in the chlorophyll b reductase reaction requires reduced ferredoxin. AB - The reduction of chlorophyllide b and its analogue zinc pheophorbide b in etioplasts of barley (Hordeum vulgare L.) was investigated in detail. In intact etioplasts, the reduction proceeds to chlorophyllide a and zinc pheophorbide a or, if incubated together with phytyldiphosphate, to chlorophyll a and zinc pheophytin a, respectively. In lysed etioplasts supplied with NADPH, the reduction stops at the intermediate step of 7(1)-OH-chlorophyll(ide) and Zn-7(1) OH-pheophorbide or Zn-7(1)-OH-pheophytin. However, the final reduction is achieved when reduced ferredoxin is added to the lysed etioplasts, suggesting that ferredoxin is the natural cofactor for reduction of chlorophyll b to chlorophyll a. The reduction to chlorophyll a requires ATP in intact etioplasts but not in lysed etioplasts when reduced ferredoxin is supplied. The role of ATP and the significance of two cofactors for the two steps of reduction are discussed. PMID- 9857046 TI - Identification of multiple Caenorhabditis elegans caspases and their potential roles in proteolytic cascades. AB - Proteases of the caspase family play a central role in the execution of programmed cell death in all metazoans examined. The Caenorhabditis elegans caspase CED-3 is essential for programmed cell death in this organism. Three additional C. elegans caspase-related genes, csp-1 (caspase homolog-1), which encodes the csp-1A, csp-1B, and csp-1C RNA species; csp-2, which encodes the csp 2A and csp-2B RNA species; and csp-3 are identified. CSP-1A, CSP-1B, CSP-2A, and CSP-2B proteins are similar in sequence to caspase proproteins. CSP-1C is similar only to large caspase subunits, and CSP-3 is similar only to small caspase subunits. CSP-1B can be activated to become a cysteine protease by processing at internal aspartate residues. Activated CSP-1B can cleave the CSP-1B, CED-3, and CSP-2B proproteins, and activated CED-3 can cleave the CED-3 and CSP-2B proproteins. Inhibitor and synthetic substrate studies further suggest that activated CSP-1B and activated CED-3 have different substrate specificities. These results suggest that C. elegans encodes several caspases that might act in proteolytic cascades to regulate processes such as programmed cell death. PMID- 9857047 TI - Novel aspects of tetramer assembly and N-terminal domain structure and function are revealed by recombinant expression of human AMP deaminase isoforms. AB - AMP deaminase isoforms purified from endogenous sources display smaller than predicted subunit molecular masses, whereas baculoviral expression of human AMPD1 (isoform M) and AMPD3 (isoform E) cDNAs produces full-sized recombinant enzymes. However, nearly 100 N-terminal amino acid residues are cleaved from each recombinant polypeptide during storage at 4 degreesC. Expression of N-truncated cDNAs (DeltaL96AMPD1 and DeltaM90AMPD3) produces stable recombinant enzymes exhibiting subunit molecular masses and kinetic properties that are similar to those reported for purified isoforms M and E. Conversely, wild type recombinant isoforms display significantly higher Km(app) values in the absence of ATP. Gel filtration analysis demonstrates native tetrameric structures for all recombinant proteins, except the wild type AMPD1 enzyme, which forms aggregates of tetramers that disperse upon cleavage of N-terminal residues at 4 degreesC. These data: 1) confirm that available literature on AMP deaminase is likely derived from N truncated enzymes and 2) are inconsistent with a new model proposing native trimeric structure of an N-truncated rabbit skeletal muscle AMP deaminase (Ranieri-Raggi, M., Montali, U., Ronca, F., Sabbatini, A., Brown, P. E., Moir, A. J. G., and Raggi, A. (1997) Biochem. J. 326, 641-648). N-terminal residues also influence actomyosin-binding properties of the enzyme, which are enhanced and suppressed by AMPD1 and AMPD3 sequences, respectively. Finally, co-expression of AMPD1 and AMPD3 recombinant polypeptides produces tetrameric enzymes with either isoform-specific or mixed subunits, and also reveals that tetramer assembly is driven by relative polypeptide abundance with no apparent preference for like subunits. PMID- 9857048 TI - Identification of a potent DNase activity associated with RNase T of Escherichia coli. AB - RNase T was first identified as an enzyme responsible for end turnover of tRNA in Escherichia coli. Its activity, specific for tRNA-C-C-A, catalyzes the release of tRNA-C-C and AMP. RNase T, along with several other RNases, plays a role in maturation of several other RNA species by a similar limited nuclease activity. In previous work, we identified the gene for RNase T, rnt, as a high copy suppressor of the UV sensitivity conferred by deficiency in three single-strand DNA-specific exonucleases, RecJ, exonuclease I, and exonuclease VII. This suggested that RNase T may process DNA substrates as well. In this work, we show that purified RNase T possesses a potent 3' to 5' single-strand DNA-specific exonucleolytic activity. Its Km for single-strand DNA substrates is many orders of magnitude lower than that for tRNA, suggesting that single-strand DNA may be a natural biological substrate for RNase T. We suggest that the DNase activity of RNase T may play a role in end trimming reactions during DNA recombination and/or DNA repair. PMID- 9857049 TI - Recombinant acyl-CoA:cholesterol acyltransferase-1 (ACAT-1) purified to essential homogeneity utilizes cholesterol in mixed micelles or in vesicles in a highly cooperative manner. AB - Acyl-coenzyme A:cholesterol acyltransferase (ACAT) is an integral membrane protein located in the endoplasmic reticulum. It catalyzes the formation of cholesteryl esters from cholesterol and long-chain fatty acyl coenzyme A. The first gene encoding the enzyme, designated as ACAT-1, was identified in 1993 through an expression cloning approach. We isolated a Chinese hamster ovary cell line that stably expresses the recombinant human ACAT-1 protein bearing an N terminal hexahistidine tag. We purified this enzyme approximately 7000-fold from crude cell extracts by first solubilizing the cell membranes with the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, then proceeding with an ACAT-1 monoclonal antibody affinity column and an immobilized metal affinity column. The final preparation is enzymologically active and migrates as a single band at 54 kDa on SDS-polyacrylamide gel electrophoresis. Pure ACAT-1 dispersed in mixed micelles containing sodium taurocholate, phosphatidylcholine, and cholesterol remains catalytically active. The cholesterol substrate saturation curves of the enzyme assayed either in mixed micelles or in reconstituted vesicles are both highly sigmoidal. The oleoyl coenzyme A substrate saturation curves of the enzyme assayed under the same conditions are both hyperbolic. These results support the hypothesis that ACAT is an allosteric enzyme regulated by cholesterol. PMID- 9857050 TI - Cse1p is required for export of Srp1p/importin-alpha from the nucleus in Saccharomyces cerevisiae. AB - In metazoan cells, the CAS protein has been shown to function as a recycling factor for the importin-alpha subunit of the classical nuclear localization signal receptor, exporting importin-alpha from the nucleus to allow its participation in multiple rounds of nuclear import. CAS is a member of a family of proteins that bear homology to the larger subunit of the nuclear localization signal receptor, importin-beta, and that are found in all eukaryotes from yeast to humans. Sequence similarity identifies the product of the Saccharomyces cerevisiae CSE1 gene as a potential CAS homologue. Here we present evidence that Cse1p is the functional homologue of CAS: Cse1p is required to prevent accumulation of Srp1p/importin-alpha in the nucleus, it localizes to the nuclear envelope in a pattern typical of nuclear transport receptors, and it associates in vivo with Srp1p in a nucleotide-specific manner. We show further that mutations in CSE1 and SRP1 have specific effects on their association and on the intracellular localization of Cse1p. PMID- 9857051 TI - Activation of p47(PHOX), a cytosolic subunit of the leukocyte NADPH oxidase. Phosphorylation of ser-359 or ser-370 precedes phosphorylation at other sites and is required for activity. AB - The leukocyte NADPH oxidase catalyzes the reduction of oxygen to superoxide (O-2) at the expense of NADPH in phagocytes and B lymphocytes. The enzyme is dormant in resting cells but becomes active when the cells are exposed to appropriate stimuli. During oxidase activation, the highly basic cytosolic oxidase component p47(PHOX) becomes phosphorylated on several serines and migrates to the plasma membrane. We report here that p47(PHOX)-deficient B lymphoblasts expressing the p47(PHOX) S359A/S370A or p47(PHOX) S359K/S370K double mutation show dramatically reduced levels of enzyme activity and phosphorylation of p47(PHOX) as compared with the same cells expressing wild type p47(PHOX). In addition, these mutant p47(PHOX) proteins fails to translocate to the plasma membrane when the cells are stimulated. In contrast, normal phosphorylation and translocation are seen in mutants containing aspartate or glutamate at positions 359 and 370, but oxidase activity is still greatly reduced. These results imply that a negative charge at position 359 and/or 370 is sufficient to allow the phosphorylation and translocation of p47(PHOX) to take place but that features unique to a phosphorylated hydroxyamino acid are required to support O-2 production. These findings, plus those from an earlier study (Inanami, O., Johnson, J. L., McAdara, J. K., El Benna, J., Faust, L. P., Newburger, P. E., and Babior, B. M. (1998) J. Biol. Chem. 273, 9539-9543), suggest that oxidase activation requires 1) the sequential phosphorylation of at least two serines on p47(PHOX): Ser-359 or Ser 370, followed by Ser-303 or Ser-304; and 2) the translocation of p47(PHOX) to the membrane at some point after the first phosphorylation takes place. PMID- 9857052 TI - A novel mechanism of CD4 down-modulation induced by monosialoganglioside GM3. Involvement of serine phosphorylation and protein kinase c delta translocation. AB - In this report the molecular mechanism(s) involved in the rapid and selective endocytosis of cell surface glycoprotein CD4 induced by exogenous monosialoganglioside GM3 in human peripheral blood lymphocytes have been investigated. Inhibition of the GM3-induced CD4 down-modulation was observed in the presence of specific protein kinase C (PKC) inhibitors. Scanning confocal microscopy revealed the translocation and clustering on the cell surface of PKC isozymes delta and theta (more evidently than alpha and beta) after GM3 treatment, suggesting the involvement of these isozymes in the ganglioside induced CD4 down-modulation. Exogenous GM3 induced phosphorylation of CD4 molecule, which then dissociated from p56(lck), as early as after 5 min. Moreover, addition of GM3 resulted in a rapid (1 min) cytosolic phospholipase A2 activation with consequent arachidonic acid release, whereas no phosphatidylinositol-phospholipase C activity was observed. Both PKC translocation and CD4 down-modulation were blocked by the trifluoromethylketone analog of arachidonic acid, a selective inhibitor of cytosolic phospholipase A2 and by mitogen-activated protein kinase inhibitor PD98059. Taken together, these findings strongly suggest that GM3 may trigger a novel mechanism of modulation of the CD4 surface expression through the activation of enzyme(s) involved in the regulation of cellular functions. PMID- 9857053 TI - Molecular mechanism of basic calcium phosphate crystal-induced activation of human fibroblasts. Role of nuclear factor kappab, activator protein 1, and protein kinase c. AB - Synovial fluid basic calcium phosphate (BCP) crystals are markers of severe joint degeneration in osteoarthritis. BCP crystals cause mitogenesis of articular cells and stimulate matrix metalloprotease production, thus promoting degradation of articular tissues. Previous work suggested that BCP crystal-induced cell activation required intracellular crystal dissolution, induction of proto oncogene expression, and activation of signal transduction pathways involving protein kinase C and mitogen-activated protein kinases. Here we further elucidate the mechanisms of BCP crystal-induced cell activation as BCP crystals activate transcription factors nuclear factor kappaB and activator protein 1 in human fibroblasts. We confirm the role of protein kinase C in BCP crystal-induced mitogenesis in human fibroblasts. In contrast, we demonstrate that BCP crystals do not activate signal transduction pathways involving protein tyrosine kinases or phosphatidylinositol 3-kinase. These data further define the mechanism of cell activation by BCP crystals and confirm its selectivity, an observation that may have therapeutic implications. PMID- 9857054 TI - Ran-dependent signal-mediated nuclear import does not require GTP hydrolysis by Ran. AB - Nuclear import of classical nuclear localization sequence-containing proteins involves the assembly of an import complex at the cytoplasmic face of the nuclear pore complex (NPC) followed by movement of this complex through the NPC and release of the import substrate into the nuclear interior. This process has historically been thought to require nucleotide hydrolysis as a source of energy. We found, using hydrolysis-resistant GTP analogs and a mutant Ran unable to hydrolyze GTP, that transport of classical nuclear localization sequence containing substrate through the NPC and release of the substrate into the nucleus did not require hydrolysis of GTP by Ran. In fact, for movement of this type of import substrate into the nuclear interior we did not observe a requirement for hydrolysis of any nucleotide triphosphate. We did, however, find that a pool of free GTP (or its structural equivalent) must be added, probably because the GDP Ran that is added must be converted to GTP Ran during the import process. We found that a requirement for GTP hydrolysis can be restored to an import mixture consisting of recombinant import factors by the addition of RCC1, the Ran guanine nucleotide exchange factor. PMID- 9857055 TI - The type I keratin 19 possesses distinct and context-dependent assembly properties. AB - Keratins (K), the cytoplasmic intermediate filament (IF) proteins of epithelial cells, are encoded by a multigene family and expressed in a tissue- and differentiation-specific manner. In human skin, keratinocytes of the basal layer of epidermis and the outer root sheath of hair follicles express K5 and K14 as their main keratins. A small subpopulation of basal cells exhibiting stem-cell like characteristics express, in addition, K19. At 40 kDa, this keratin is the smallest IF protein due to an exceptionally short carboxyl-terminal domain. We examined the assembly properties of K19 and contrasted them to K14 in vitro and in vivo. Relative to K5-K14, we find that K5-K19 form less stable tetramers that polymerize into shorter and narrower IFs in vitro. When transiently co-expressed in cultured baby hamster kidney cells, the K5 and K19 combination fails to form a filamentous array, whereas the K5-K14 and K8-K19 ones readily do so. Transient expression of K19 in the epithelial cell lines T51B-Ni and A431 results in its integration into the endogenous keratin network with minimal if any perturbation. Collectively, these results indicate that K19 possesses assembly properties that are distinct from those of K14 and suggest that it may impart unique properties to the basal cells expressing it in skin epithelia. PMID- 9857056 TI - c-Src activation plays a role in endothelin-dependent hypertrophy of the cardiac myocyte. AB - Activation of the atrial natriuretic peptide (ANP) gene is regarded as one of the earliest and most reliable markers of hypertrophy in the ventricular cardiac myocyte. We have examined the role of the nonreceptor tyrosine kinases in the signaling mechanism(s) leading to hypertrophy using human ANP gene promoter activity as a marker. Endothelin (ET), a well known hypertrophic agonist, increased activity of c-Src, c-Yes, and Fyn within minutes and promoted a selective redistribution of each of these kinases within the cell. Overexpression of c-Src effected a significant increase in activity of a cotransfected human ANP promoter-driven chloramphenicol acetyl transferase reporter, while expression of either c-Yes or Fyn was considerably less effective in this regard. ET-dependent stimulation of the human ANP gene promoter was partially inhibited by co transfection with dominant negative Ras or dominant negative Src or Csk or by treatment with the potent Src family-selective tyrosine kinase inhibitor PP1, suggesting that the Src family kinases are involved in signaling ET-dependent activation of this promoter. Both ET- and Src-dependent activation of the ANP promoter required the presence of a CArG motif in a serum response element-like structure between -422 and -413 but did not appear to require assembly of a ternary complex for full activity. These findings support a role for Src in the activation of ANP gene expression and suggest that this kinase may contribute in an important way to the signaling mechanisms that activate hypertrophy in the cardiac myocyte. PMID- 9857057 TI - Hop as an adaptor in the heat shock protein 70 (Hsp70) and hsp90 chaperone machinery. AB - Hop, an abundant and conserved protein of unresolved function, binds concomitantly with heat shock protein 70 (Hsp70) and Hsp90, participates with heat shock proteins at an intermediate stage of progesterone receptor assembly, and is required for efficient assembly of mature receptor complexes in vitro. A largely untested hypothesis is that Hop functions as an adaptor that targets Hsp90- to Hsp70-substrate complexes; if true, then loss of either Hsp70 binding or Hsp90 binding by Hop should equally disrupt its ability to promote assembly of mature receptor complexes. To generate Hop mutants that selectively disrupt heat shock protein interactions, highly conserved amino acids in the previously mapped Hsp70 and Hsp90 binding domains of Hop and in a conserved C-terminal domain were targeted for small substitutions and deletions. In co-precipitation assays, these mutants displayed selective loss of association with heat shock proteins. In assays using Hop-depleted rabbit reticulocyte lysate for the cell-free assembly of receptor complexes, none of the Hop mutants inhibited Hsp70 binding to receptor, but all mutants were defective in supporting Hsp90-receptor interactions. Thus, Hop has a novel role in the chaperone machinery as an adaptor that can integrate Hsp70 and Hsp90 interactions. PMID- 9857058 TI - Degradation of proto-oncoprotein c-Rel by the ubiquitin-proteasome pathway. AB - The c-rel proto-oncogene product, c-Rel, belongs to the Rel/NF-kappaB transcription factor family, which regulates a large variety of cellular functions. The activation of NF-kappaB involves the degradation of the inhibitor, IkappaB, through the ubiquitin-proteasome (Ub-Pr)-mediated pathway. Here we report that the turnover of c-Rel is also regulated by the Ub-Pr pathway, thus adding another level of complexity to the regulation of NF-kappaB. High molecular weight ubiquitinated c-Rel conjugates are detected in cells and accumulate in cells treated with proteasome inhibitors. In a cell-free in vitro degradation assay, c-Rel is degraded specifically through the Ub-Pr pathway. N-terminally truncated c-Rel is readily degraded, implying the dispensability of N-terminal sequence; in contrast, a series of deletion mutants missing C-terminal sequences display a reduced susceptibility to the degradation. Interestingly, the sequence between residues 118 and 171 of c-Rel, i.e. the region immediately following the c-Rel/v-Rel homology domain, appears to play an important role in mediating ubiquitin conjugation and the subsequent degradation. Together with our previous study showing an elevated tumorigenic potential for C-terminally truncated mutants, our data suggest that the C-terminal domain of c-Rel plays an important role in mediating c-Rel degradation and growth control. PMID- 9857059 TI - Transcriptional up-regulation of the delayed early gene HRS/SRp40 during liver regeneration. Interactions among YY1, GA-binding proteins, and mitogenic signals. AB - Arg-Ser-rich domain-containing proteins (SR proteins), a family of splicing factors, can regulate pre-mRNA alternative splicing in a concentration dependent manner. Thus, the relative expression of various SR proteins may play an important role in alternative splicing regulation. HRS/SRp40, an SR protein and delayed early gene in liver regeneration, can mediate alternative splicing of fibronectin mRNA. Here we determined that transcription of the HRS/SRp40 gene is induced about 5-fold during liver regeneration, similar to the level of steady state mRNA. We found that both mouse and human HRS promoters lack TATA and CAAT boxes. The mouse promoter region from -130 to -18, which contains highly conserved GA-binding protein (GABP) and YY1 binding sites, conferred high transcriptional activity. While GABPalpha/GABPbeta heterodimer transactivated the HRS promoter, YY1 functioned as a repressor. During liver regeneration, the relative amount of GABPalpha/GABPbeta heterodimer increased 3-fold, and YY1 changed little, which could partially account for the increase in HRS gene transcription. Interleukin-6, a critical mitogenic component of liver regeneration, was able to relieve the repressive activity of the YY1 site within the HRS promoter. The combined effect of small changes in the level of existing transcription factors and mitogenic signals may explain the transcriptional activation of the HRS gene during cell growth. PMID- 9857060 TI - Translocation efficiency, susceptibility to proteasomal degradation, and lipid responsiveness of apolipoprotein B are determined by the presence of beta sheet domains. AB - Apolipoprotein (apo) B100 is an atypical secretory protein in that its translocation across the endoplasmic reticulum membrane is inefficient, resulting in the partial translocation and exposure of apoB100 on the cytoplasmic surface of the endoplasmic reticulum. Cytosolic exposure leads to the association of nascent apoB with heat shock protein 70 and to its predisposition to ubiquitination and proteasomal degradation. The basis for the inefficient translocation of apoB100 remains unclear and controversial. To test the hypothesis that beta sheet domains present in apoB100 contribute to its inefficient translocation, we created human apoB chimeric constructs apoB13,16 and apoB13,13,16, which contain amino-terminal alpha globular domains but no beta sheet domains, and apoB13,16,beta, which has an amphipathic beta sheet domain of apoB100 inserted into apoB13,16. These constructs, along with carboxyl-terminal truncations of apoB100, apoB34 and apoB42, were used to transfect HepG2 and Chinese hamster ovary cells. In contrast to the lack of effect of proteinase K on apoB13,16 and apoB13,13,16, the levels of apoB34, apoB42, and apoB13,16,beta were decreased by 70-85% after proteinase K-induced proteolysis in both HepG2 and Chinese hamster ovary cells. Either oleic acid or proteasomal inhibitors (N acetyl-leucinyl-leucinyl-norleucinal and lactacystin) significantly increased the cell levels of apoB13,16,beta, apoB34, apoB42, and full-length apoB100 but had no effect on the cell levels of apoB13,16 and apoB13,13,16. When HepG2 cells were incubated with a microsomal triglyceride transfer protein inhibitor, the cellular levels of apoB13,16,beta, apoB34, and apoB42 were decreased by 70-80%, whereas the levels of apoB13,16 and apoB13,13,16 were unaffected. The effects of microsomal triglyceride transfer protein inhibition were reversed by lactacystin. Our results clearly demonstrate that the translocation efficiency, susceptibility to proteasomal degradation, and lipid responsiveness of apoB were determined by the presence of a lipid binding beta sheet domain. It is possible that beta sheet domains may at least transiently facilitate the interaction of apoB with the lipid bilayer surrounding the translocation channel. PMID- 9857061 TI - A novel F box protein, NFB42, is highly enriched in neurons and induces growth arrest. AB - NFB42 (neural F Box 42 kDa) is a novel gene product that is highly enriched in the nervous system. Its predicted protein contains an F box, a motif recently shown to couple cell cycle regulation to the proteasome pathway (Bai, C., Sen, P., Hofmann, K., Ma, L., Goebl, M., Harper, J. W., and Elledge, S. (1996) Cell 86, 263-274). NFB42 mRNA and protein are expressed in all major areas of the adult rat brain but are not detected in non-neural tissues. NFB42 protein is localized primarily to the cytoplasm of neurons and does not appear to be present in glia. The presence of an F box in NFB42 suggests that it may be involved in cell cycle regulation; however, its expression in postmitotic neurons indicates that it is not involved in regulating typical cell cycle events. In an initial attempt to characterize the function of this protein, NFB42 was transfected into N1E-115 neuroblastoma and Chinese hamster ovary cells. The expression of full length NFB42, but not an F box deletion mutant, inhibits proliferation in both cell lines. Additional experiments demonstrate that NFB42 interacts with Skp1p, a component of the proteasome pathway, and deletion of the F box also inhibits this interaction. Overall, the expression pattern of NFB42, along with the presence of an F box domain and the ability to inhibit growth, suggests that it may play a role in maintaining neurons in a postmitotic state. PMID- 9857062 TI - Post-transcriptional regulation of a milk membrane protein, the sialomucin complex (Ascites sialoglycoprotein (ASGP)-1/ASGP-2, rat muc4), by transforming growth factor beta. AB - Sialomucin complex (SMC, Rat Muc4) is a heterodimeric glycoprotein complex consisting of a mucin subunit ASGP-1 (ascites sialoglycoprotein-1) and a transmembrane subunit ASGP-2, which can act as a ligand for the receptor tyrosine kinase ErbB2. SMC is highly expressed on the surface of ascites 13762 rat mammary adenocarcinoma cells, approximately 100 times the level in lactating mammary gland and 10(4) times that in virgin mammary gland. SMC is sharply increased at mid-pregnancy in a manner similar to beta-casein. Unlike beta-casein, SMC appears to be regulated post-transcriptionally. Its transcript is present in both virgin and pregnant mammary tissue, and SMC synthesis is induced rapidly in cultured primary mammary epithelial cells from either normal pregnant or virgin rats. SMC protein, but not transcript, levels are significantly reduced when mammary cells are cultured in Matrigel, a reconstituted basement membrane which stimulates casein expression. SMC precursor is synthesized in Matrigel at a 10-fold lower rate. Matrigel has no effect on either the level of SMC or its transcript in cultured 13762 mammary tumor cells. The Matrigel effect on primary mammary and 13762 cells is mimicked by transforming growth factor beta, a component associated with this complex matrix. These results indicate that SMC is a novel product of normal mammary gland and milk, which is post-transcriptionally regulated by transforming growth factor beta in normal mammary gland, but not in 13762 mammary adenocarcinoma cells. PMID- 9857063 TI - Degradation of the G protein-coupled receptor kinase 2 by the proteasome pathway. AB - GRK2 is a ubiquitous member of the G protein-coupled receptor kinase (GRK) family and has been shown to play a key role in determining the desensitization and resensitization patterns of a variety of G protein-coupled receptors. In this report, we show that GRK2 is actively degraded by the proteasome proteolytic pathway, unveiling a new mechanism for the rapid regulation of its expression levels. Interestingly, activation of beta2-adrenergic receptors (beta2AR) markedly increases GRK2 ubiquitination and degradation through the proteasome pathway. In addition, blocking GRK2 degradation notably alters beta2AR signaling and internalization, consistent with a relevant physiological role for GRK2 proteasomal degradation. Activity-dependent modulation of GRK2 cellular levels emerges as an important mechanism for modulating the cellular response to agonists acting through G protein-coupled receptors. PMID- 9857064 TI - Structure-function studies of human leptin. AB - To elucidate the structural requirement of human leptin for its functions, the wild-type, mutant-type, C-terminal deletion, and N-terminal deletion were expressed in Escherichia coli and purified in soluble forms. These leptin analogs were intracerebroventrically injected into C57BL/6J ob/ob mice, and their in vivo biological activities were evaluated. The mutant-type leptin lacking a C-terminal disulfide bond reduced food intake at doses of more than 15 pmol/mouse, which was as effective as the wild-type leptin. C-terminal deletion without the loop structure, also significantly, but to a lesser extent, reduced food intake at doses of more than 90 pmol/mouse. However, N-terminal deletions showed no effect on food intake. We also evaluated the effects of the leptin analogs on radiolabeled leptin binding to its receptor in the choroid plexus using autoradiography. An excess of unlabeled mutant-type leptin as well as wild-type leptin led to complete inhibition of binding. C-terminal deletions led to weak inhibitory activity, whereas N-terminal deletions caused no inhibitory activity. These results clearly demonstrate that the N-terminal region of leptin is essential for both its biological and receptor binding activities. The amino acid sequence of the C-terminal loop structure is also important for enhancing these actions, whereas the C-terminal disulfide bond is not needed. PMID- 9857065 TI - Effect of transmembrane and kinase domain mutations on fibroblast growth factor receptor 3 chimera signaling in PC12 cells. A model for the control of receptor tyrosine kinase activation. AB - The effect of six point mutations causing various human skeletal dysplasias, occurring in the transmembrane (TM) and kinase domains (KD) of fibroblast growth factor receptor 3, were introduced into a chimera composed of the extracellular domain of human platelet-derived growth factor beta and the TM and intracellular domains of hFGFR3. Stable transfectants in rat PC12 cells showed distinct differences in the two classes of mutations. The cells containing TM mutants displayed normal expression and activation but higher responsiveness to lower doses of ligand. The KD mutants showed significantly altered expression patterns. Normal amounts of a lower Mr receptor (p130) reflecting incomplete glycosylation, but only greatly decreased amounts of the mature (p170) form, were observed. However, the latter material showed normal ligand-dependent activation. In contrast, the p130 form, which is regularly observed in the expression of both native and chimeric receptors, exhibits strong ligand-independent tyrosine phosphorylation, particularly with the K650E mutation. Expression of two of the KD mutants (K650M and K650E), under control of an inducible metallothionein promoter, indicated that this receptor was sufficiently autoactivated to produce at least partial differentiation and, in the case of the K650E mutation, to induce ligand-independent neurite outgrowth. A model is presented that suggests that the low Mr (p130) KD mutants can, under the right conditions, signal intracellularly, but when they are fully glycosylated and move to the cell surface they adopt a normal, inhibited conformation, in the form of ligand independent dimers, that neutralizes the effects of the mutations. When ligands bind, these dimeric receptors are activated in a normal manner. This model suggests that unliganded dimers may be a common intermediate in receptor tyrosine kinase signaling. PMID- 9857066 TI - Role of the COOH-terminal domains of meprin A in folding, secretion, and activity of the metalloendopeptidase. AB - Secreted forms of the alpha subunit of recombinant mouse meprin A include an NH2 terminal prosequence, a catalytic domain, and three COOH-terminal domains designated as MAM (meprin, A-5 protein, receptor protein-tyrosine phosphatase mu), MATH (meprin and TRAF homology), and AM (after MATH). In this study, the importance of these COOH-terminal domains for biosynthesis of secreted, activable forms of the protease was investigated. Transcripts of the meprin subunit truncated after the protease (alpha(1-275)), MAM (alpha(1-452)), and MATH (alpha(1-528)) domains or with individual domains deleted (DeltaMAM, DeltaMATH, and DeltaAM), were transfected into human embryonic kidney 293 cells. The wild type subunit, DeltaMATH, DeltaAM, alpha(1-452), and alpha(1-528) were secreted into the media, although the DeltaAM mutant was secreted at very low levels. The DeltaMATH and alpha(1-452) mutants were not activable by limited proteolysis. The alpha(1-528) mutant was as active as wild-type meprin alpha against a bradykinin substrate, but had no activity against azocasein, and it, as all other mutants, was more vulnerable to extensive degradation by proteases than the wild-type protein. Pulse-chase experiments revealed that the DeltaMAM and alpha(1-275) mutants were rapidly degraded within cells. Treatment with lactacystin, a specific inhibitor of the proteasome, significantly decreased the degradation, indicating that the mutants lacking the MAM domain are degraded by the proteasome as misfolded proteins. These results indicate that the MAM domain is necessary for correct folding and transport through the secretory pathway, the MATH domain is required for folding of an activable zymogen, and the AM domain is important for activity against proteins and efficient secretion of the protein. The work demonstrates the interdependence of the domains for correct folding of an activable, stable, mature enzyme. PMID- 9857067 TI - A toxic fusion protein accumulating between the mitochondrial membranes inhibits protein assembly in vivo. AB - When overexpressed in Saccharomyces cerevisiae, beta-galactosidase fusion proteins directed to the mitochondria are toxic, preventing growth of yeast cells on non-fermentable carbon sources (Emr, S. D., Vassarotti, A., Garrett, J., Geller, B. L., Takeda, M., and Douglas, M. G. (1986) J. Cell Biol. 102, 523-533). We show that such fusion proteins interfere with the assembly of respiratory complexes in the mitochondrial inner membrane, without blocking protein translocation. The gene YME1, encoding an ATP-dependent metalloprotease of the mitochondrial inner membrane, acts as a suppressor of this defect; a 3-fold overexpression of Yme1p is sufficient to restore respiratory complex assembly and mitochondrial function. Detailed knowledge of the topology and effect of the toxic beta-galactosidase fusion proteins will permit the identification and characterization of components that control protein sorting and protein assembly within the mitochondrial inner membrane. PMID- 9857068 TI - Cbl-mediated negative regulation of the Syk tyrosine kinase. A critical role for Cbl phosphotyrosine-binding domain binding to Syk phosphotyrosine 323. AB - The proto-oncogene product Cbl has emerged as a potential negative regulator of the Syk tyrosine kinase; however, the nature of physical interactions between Cbl and Syk that are critical for this negative regulation remains unclear. Here we show that the phosphotyrosine-binding (PTB) domain within the N-terminal transforming region of Cbl (Cbl-N) binds to phosphorylated Tyr323 in the linker region between the Src homology 2 and kinase domains of Syk, confirming recent results by another laboratory using the yeast two-hybrid approach (Deckert, M., Elly, C., Altman, A., and Liu, Y. C. (1998) J. Biol. Chem. 273, 8867-8874). A PTB domain-inactivating point mutation (G306E), corresponding to a loss-of-function mutation in the Caenorhabditis elegans Cbl homologue SLI-1, severely compromised Cbl-N/Syk binding in vitro and Cbl/Syk association in transfected COS-7 cells. Using heterologous expression in COS-7 cells, we investigated the role of Cbl PTB domain binding to Syk Tyr323 in the negative regulation of Syk. Co-expression of Cbl with Syk in COS-7 cells led to a dose-dependent decrease in the autophosphorylated pool of Syk and in phosphorylation of an in vivo substrate, CD8-zeta. Unexpectedly, these effects were largely due to the loss of Syk protein. Both the decrease in Syk and CD8-zeta phosphorylation and reduction in Syk protein levels were blocked by either G306E mutation in Cbl or by Y323F mutation in Syk. These results demonstrate a critical role for the Cbl PTB domain in the recruitment of Cbl to Syk and in Cbl-mediated negative regulation of Syk. PMID- 9857070 TI - Tyrosine phosphorylation of syndecan-1 and -4 cytoplasmic domains in adherent B82 fibroblasts. AB - The syndecans, a family of cell surface proteoglycans, have highly conserved cytoplasmic domains that bind proteins containing PDZ domains and co-localize with the actin cytoskeleton. The syndecan cytoplasmic domains contain four conserved tyrosine residues, two of which are located within favorable sequences for phosphorylation. Endogenous tyrosine phosphorylation of syndecans-1 and -4 is detected in adherent B82 fibroblasts. Approximately 1.5% of total syndecan is endogenously phosphorylated, while most, if not all, cell surface syndecan is phosphorylated following treatment with the tyrosine phosphatase inhibitor pervanadate. Syndecan phosphorylation is also detected in Raji-S1 and NMuMG cells, but only following treatment with vanadate or pervanadate, suggesting that endogenous phosphorylation is maintained in an "off" state in these cells. Endogenous syndecan phosphorylation in B82 cells is rapidly blocked by genistein (IC50 < 10 microM) confirming the presence of a constitutively active kinase and a corresponding tyrosine phosphatase. Phosphorylation is also inhibited by herbimycin A (IC50 < 1.0 microM) and staurosporine (IC50 < 1.0 nM), suggesting a role for Src family kinases in regulating syndecan phosphorylation. Together, these data suggest an important role for tyrosine phosphorylation of the syndecan cytoplasmic domains in regulating downstream signaling events in response to cell adhesion and/or growth factor activity. PMID- 9857069 TI - Cloning and characterization of a novel mammalian PP2C isozyme. AB - PP2C is a structurally diversified protein phosphatase family with a wide range of functions in cellular signal transduction. A novel PP2C subtype, designated PP2Cdelta, was identified from a rat cDNA clone, which encodes a protein of 392 amino acid residues. While PP2Cdelta shares approximately 30% sequence identity in its catalytic domain with the mammalian PP2C, it lacks a 90-residue carboxyl terminal sequence conserved in mammalian PP2C. Northern blot analysis showed that PP2Cdelta is widely expressed in rat tissues. The transcription of the PP2Cdelta gene was activated in response to stress, such as the addition of ethanol to the culture medium or UV irradiation of cells. Recombinant PP2Cdelta purified from bacteria exhibited a potent Mn2+-dependent serine/threonine phosphatase activity. Unlike other members of the PP2C family, the activity of PP2Cdelta was inhibited, rather than stimulated, by Mg2+. Transfection with PP2Cdelta resulted in inhibition of cell growth, precluding generation of stable 293 or CHO transfectants. Using a modified tetracycline-regulated PP2Cdelta-GFP dicistronic expression cassette, it was revealed that overexpression of PP2Cdelta blocked cell cycle progression and arrested cells at early S phase, resulting in inhibition of DNA synthesis and leading to cell death. These results suggest that PP2Cdelta plays a role in regulation of cell cycle progression via dephosphorylation of its substrates whose appropriate phosphorylation states might be crucial for cell proliferation. PMID- 9857072 TI - Equilibrium binding studies of non-claret disjunctional protein (Ncd) reveal cooperative interactions between the motor domains. AB - Non-claret disjunctional protein (Ncd) is a minus end-directed microtubule motor required for normal spindle assembly and integrity during Drosophila oogenesis. We have pursued equilibrium binding experiments to examine the affinity of Ncd for microtubules in the presence of the ATP nonhydrolyzable analog 5'-adenylyl beta, gamma-imidodiphosphate (AMP-PNP), ADP, or ADP + Pi using both dimeric (MC1) and monomeric (MC6) Ncd constructs expressed in Escherichia coli. Both MC1 and MC6 sediment with microtubules in the absence of added nucleotide as well as in the presence of either ADP or AMP-PNP. Yet, in the presence of ADP + Pi, there is a decrease in the affinity of both MC1 and MC6 for microtubules. The data for dimeric MC1 show that release of the dimer to the supernatant is sigmoidal with the apparent Kd(Pi) for the two phosphate sites at 23.3 and 1.9 mM, respectively. The results indicate that binding at the first phosphate site enhances binding at the second site, thus cooperatively stimulating release. Stopped-flow kinetics indicate that MgATP promotes dissociation of the Mt.MC1 complex at 14 s-1, yet AMP-PNP has no effect on the Mt.MC1 complex. These results are consistent with a model for the ATPase cycle in which ATP hydrolysis occurs on the microtubule followed by detachment as the Ncd.ADP.Pi intermediate. PMID- 9857071 TI - Nuclear sterol regulatory element-binding proteins activate genes responsible for the entire program of unsaturated fatty acid biosynthesis in transgenic mouse liver. AB - Previous studies have shown that the rate of fatty acid synthesis is elevated by more than 20-fold in livers of transgenic mice that express truncated nuclear forms of sterol regulatory element-binding proteins (SREBPs). This was explained in part by an increase in the levels of mRNA for the two major enzymes of fatty acid synthesis, acetyl-CoA carboxylase and fatty acid synthase, whose transcription is stimulated by SREBPs. Fatty acid synthesis also requires a source of acetyl-CoA and NADPH. In the current studies we show that the levels of mRNA for ATP citrate lyase, the enzyme that produces acetyl-CoA, are also elevated in the transgenic livers. In addition, we found marked elevations in the mRNAs for malic enzyme, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase, all of which produce NADPH. Finally, we found that overexpressing two of the SREBPs (1a and 2) led to elevated mRNAs for stearoyl-CoA desaturase 1 (SCD1), an isoform that is detectable in nontransgenic livers, and SCD2, an isoform that is not detected in nontransgenic livers. This stimulation led to an increase in total SCD activity in liver microsomes. Together, all of these changes would be expected to lead to a marked increase in the concentration of monounsaturated fatty acids in the transgenic livers, and this was confirmed chromatographically. We conclude that expression of nuclear SREBPs is capable of activating the entire coordinated program of unsaturated fatty acid biosynthesis in mouse liver. PMID- 9857073 TI - AppA, a redox regulator of photosystem formation in Rhodobacter sphaeroides 2.4.1, is a flavoprotein. Identification of a novel fad binding domain. AB - The AppA protein is required for increased photosystem gene expression upon transition of the facultatively photoheterotrophic bacterium Rhodobacter sphaeroides 2.4.1 from aerobic to anaerobic photosynthetic conditions. AppA shows no obvious similarity to proteins with established function. Genetic evidence suggests that its effect is exerted through modulation of the activity of the repressor PpsR, which controls expression of multiple photosystem genes. To gain insight into the nature of AppA involvement in redox-dependent photosystem gene expression, the appA gene was overexpressed in Escherichia coli. AppA was produced as insoluble inclusion bodies. The purified inclusion bodies were found to contain FAD. By overexpressing various deletion derivatives, we were able to localize the region of AppA sufficient for FAD binding to approximately 120 amino terminal residues. To assess the role of FAD binding in AppA function, we constructed an AppA derivative lacking the entire FAD binding domain. Surprisingly, this derivative complemented the AppA null mutant undergoing transition from aerobic to anaerobic photosynthetic growth conditions almost to the same extent as the full-length AppA protein. When the sequence of the amino terminal portion of AppA was examined, it was shown not to contain any known flavin binding motifs. However, two open reading frames of unknown function, showing significant similarity to the amino terminus of AppA, were identified, i.e. Synechocystis sp. Srl1694 and E. coli F403. The latter gene was amplified and overexpressed in E. coli, and the partially purified F403 protein was found to contain FAD as a cofactor. We have therefore concluded that the amino terminus of AppA represents a novel FAD binding domain present in a small group of bacterial proteins. The binding of FAD by AppA may be the first clue as to how this regulatory protein is involved in redox-regulated reactions. PMID- 9857074 TI - Down-regulation of human granzyme B expression by glucocorticoids. Dexamethasone inhibits binding to the Ikaros and AP-1 regulatory elements of the granzyme B promoter. AB - The serine protease granzyme B is an essential component of the granule exocytosis pathway, a major apoptotic mechanism used by cytotoxic T lymphocytes and natural killer cells to induce target cell apoptosis. Granzyme B gene transcription is induced in activated lymphocytes upon antigenic stimulation, and several regulatory regions including CBF, AP-1, and Ikaros binding sites have been shown to be essential in the control of granzyme B promoter activation. Dexamethasone, a glucocorticoid that is widely used as an immunomodulatory and anti-inflammatory agent, inhibits granzyme B mRNA transcript in phytohemagglutinin-activated peripheral blood mononuclear cells. Transfection of a reporter construct containing the -148 to +60 region of the human granzyme B promoter demonstrated that this region was the target for dexamethasone repression. Mutation of Ikaros or AP-1 binding sites in the context of the granzyme B promoter demonstrated that both sites participate in dexamethasone mediated inhibition of the granzyme B promoter activity. Electromobility shift assay revealed that dexamethasone abolished the binding of nuclear transcription factors to the Ikaros binding site and reduced AP-1 binding activity. These results indicate that dexamethasone is able to abrogate the transcriptional activity of the human granzyme B gene promoter by inhibiting the binding of nuclear factors at the AP-1 and Ikaros sites. PMID- 9857075 TI - Neisseria gonorrhoeae porin modulates phagosome maturation. AB - The porin (PorB) of Neisseria gonorrhoeae has been implicated in the pathogenesis of this species. Porin is believed to translocate from the bacterial outer membrane into target cell membranes affecting various cell functions. Here we investigated the effect of porin on phagosome maturation. Phagocytosis of latex beads by human macrophages was allowed in the presence or absence of purified porin. Isolation of latex bead-containing phagosomes and subsequent two dimensional gel electrophoresis revealed substantial differences in the phagosomal protein composition. Immunoblotting detected higher amounts of annexin II and the early endocytic markers Rab5 and transferrin receptor and decreased levels of the late endocytic markers Rab7 and cathepsin D in phagosomes obtained in the presence of porin compared with those obtained in its absence. Furthermore, association of Rab4 with the latex bead-containing phagosomes was revealed by flow cytometry. The amount of this small GTPase was markedly higher in the phagosomes isolated in the presence of porin. The data thus indicate that neisserial porin is itself able to arrest phagosome maturation within macrophages. PMID- 9857076 TI - Identification of structural elements involved in the interaction of simian virus 40 small tumor antigen with protein phosphatase 2A. AB - SV40 small tumor antigen (small-t) was used as a model to identify structural elements involved in the interactions between regulatory proteins and protein phosphatase 2A (PP2A). Using mutant proteins and synthetic peptides, we identified a small domain within small-t that is a major site for interaction with the dimeric form of PP2A. A series of small-t truncation mutants identified a region surrounding the first of two conserved cysteine clusters that was critical for interaction with PP2A. These mutants also identified additional regions of small-t that contribute to high affinity interaction. Deletion of residues 110-119, which encompass the first cysteine cluster, resulted in a protein that failed to bind to PP2A. Synthetic peptides that contained residues 105-122 of small-t blocked binding of small-t to PP2A. These peptides also inhibited the phosphatase activity of PP2A in a manner analogous to full-length small-t. The active small-t peptides adopt a beta-strand structure that was essential for high affinity interaction with the PP2A dimer. Based on circular dichroism measurements, the same cysteine cluster-containing peptides that bind to PP2A also interact with zinc. Interaction with zinc required the conserved cysteines but was not required for interaction with PP2A. PMID- 9857077 TI - Functional domains of the alpha1 catalytic subunit of the AMP-activated protein kinase. AB - The AMP-activated protein kinase is a heterotrimeric enzyme, important in cellular adaptation to the stress of nutrient starvation, hypoxia, increased ATP utilization, or heat shock. This mammalian enzyme is composed of a catalytic alpha subunit and noncatalytic beta and gamma subunits and is a member of a larger protein kinase family that includes the SNF1 kinase of Saccharomyces cerevisiae. In the present study, we have identified by truncation and site directed mutagenesis several functional domains of the alpha1 catalytic subunit, which modulate its activity, subunit association, and protein turnover. C terminal truncation of the 548-amino acid (aa) wild-type alpha1 protein to aa 312 or 392 abolishes the binding of the beta/gamma subunits and dramatically increases protein expression. The full-length wild-type alpha1 subunit is only minimally active in the absence of co-expressed beta/gamma, and alpha1(1-392) likewise has little activity. Further truncation to aa 312, however, is associated with a large increase in enzyme specific activity, thus revealing an autoinhibitory sequence between aa 313 and 392. alpha-1(1-312) still requires the phosphorylation of the activation loop Thr-172 for enzyme activity, yet is now independent of the allosteric activator, AMP. The increased levels of protein expression on transient transfection of either truncated alpha subunit cDNA are because of a decrease in enzyme turnover by pulse-chase analysis. Taken together, these data indicate that the alpha1 subunit of AMP-activated protein kinase contains several features that determine enzyme activity and stability. A constitutively active form of the kinase that does not require participation by the noncatalytic subunits provides a unique reagent for exploring the functions of AMP-activated protein kinase. PMID- 9857078 TI - The NH2-terminal region of apolipoprotein B is sufficient for lipoprotein association with glycosaminoglycans. AB - An initial event in atherosclerosis is the retention of lipoproteins within the intima of the vessel wall. The co-localization of apolipoprotein (apo) B and proteoglycans within lesions has suggested that retention is due to lipoprotein interaction with these highly electronegative glycoconjugates. Both apoB100- and apoB48-containing lipoproteins, i.e. low density lipoproteins (LDLs) and chylomicron remnants, are atherogenic. This suggests that retention is due to determinants in the initial 48% of apoB. To test this, the interaction of an apoB fragment (apoB17), and apoB48- and apoB100- containing lipoproteins with heparin, subendothelial matrix, and artery wall purified proteoglycans was studied. ApoB100-containing LDL from humans and human apoB transgenic mice and apoB48 containing LDLs from apoE knockout mice were used. Despite the lack of the carboxyl-terminal 52% of apoB, the apoB48-LDL bound to heparin-affinity gel as well as did apoB100-LDL. An NH2-terminal fragment containing 17% of full-length apoB was made using a recombinant adenovirus; apoB17 bound to heparin as well as did LDL. Monoclonal antibodies against the NH2-terminal region of apoB decreased apoB100 LDL binding to heparin, whereas antibodies against the LDL receptor binding region did not alter LDL-heparin interaction. The role of the NH2 terminal region of apoB in LDL interaction with matrix molecules was also assessed. Media containing apoB17 decreased LDL binding to subendothelial matrix by 42%. Moreover, removal of the apoB17 by immunoprecipitation abrogated the inhibitory effect of these media. Antibodies to the NH2-terminal region decreased LDL binding to matrix and dermatan sulfate proteoglycans. Purified apoB17 effectively competed for binding of LDL to artery derived decorin and to subendothelial matrix. Thus, despite the presence of multiple basic amino acids near the LDL receptor-binding domain of LDL, the NH2-terminal region of apoB is sufficient for the interaction of lipoproteins with glycoconjugates produced by endothelial and smooth muscle cells. The presence of a proteoglycan-binding site in the NH2-terminal region of apoB may explain why apoB48- and apoB100-containing lipoproteins are equally atherogenic. PMID- 9857079 TI - The 72-kDa component of signal recognition particle is cleaved during apoptosis. AB - Proteins cleaved by apoptotic caspases are commonly recognized by autoantibodies found in the serum of patients with rheumatic disease. We report that the 72-kDa signal recognition particle (SRP) protein, a rare target of autoantibodies found in the serum of patients with dermatomyositis and systemic lupus erythematosus, is rapidly cleaved in Jurkat T cells treated with apoptotic (i.e. Fas ligation, treatment with gamma or ultraviolet radiation, or co-culture with anisomycin or staurosporine) but not proliferative (CD3 cross-linking) stimuli. Cleavage of SRP 72 produces a 66-kDa amino-terminal fragment and a 6-kDa carboxyl-terminal fragment that is selectively phosphorylated on serine residues. Cleavage of SRP 72 is prevented by chemical and peptide caspase inhibitors, and by overexpression of bcl-2, an inhibitor of apoptotic cell death. Analysis of the carboxyl terminus of SRP 72 has identified a putative cleavage site (SELD/A) for group III caspases, and carboxyl-terminal serine residues that are highly conserved in phylogeny. Both serine phosphorylation and caspase cleavage of SRP 72 are observed in cells derived from human, dog, rat, and mouse. Canine SRP 72 is cleaved in vitro by recombinant caspase 3 but retains the ability to mediate transport of a signal peptide-containing protein into the endoplasmic reticulum lumen. The 72-kDa component of the SRP joins a growing list of autoantigens that undergo post-translational modifications during programmed cell death. PMID- 9857080 TI - Bacterial lipopolysaccharide disrupts endothelial monolayer integrity and survival signaling events through caspase cleavage of adherens junction proteins. AB - Bacterial lipopolysaccharide or endotoxin induces actin reorganization, increased paracellular permeability, and endothelial cell detachment from the underlying extracellular matrix in vitro. We studied the effect of endotoxin on transendothelial albumin flux and detachment of endothelial cells cultured on gelatin-impregnated filters. The endotoxin-induced changes in endothelial barrier function and detachment occurred at doses and times that were compatible with endotoxin-induced apoptosis. Since the actin cytoskeleton and cell-cell and cell matrix adhesion all participate in the regulation of the paracellular pathway and cell-matrix interactions, we studied whether protein components of the actin linked adherens junctions were modified in response to endotoxin. Components of cell-cell (beta- and gamma-catenin) and cell-matrix (focal adhesion kinase and p130(Cas)) adherens junctions were cleaved by caspases activated during apoptosis with dose and time requirements that paralleled those seen for barrier dysfunction and detachment. Cleavage of focal adhesion kinase led to its dissociation from the focal adhesion-associated signaling protein, paxillin, resulting in reduced paxillin tyrosine phosphorylation. Inhibition of caspase mediated cleavage of these proteins protected against detachment but not opening of the paracellular pathway. Therefore, endotoxin-induced disruption of endothelial monolayer integrity and survival signaling events is mediated, in part, through caspase cleavage of adherens junction proteins. PMID- 9857081 TI - Substrate specificity and kinetic mechanism of the insect sulfotransferase, retinol dehydratase. AB - Spodoptera frugiperda retinol dehydratase catalyzes the conversion of retinol to the retro-retinoid anhydroretinol. It shares sequence homology with the family of mammalian cytosolic sulfotransferases and provides the first link between sulfotransferases and retinol metabolism. In this study the enzymatic properties of retinol dehydratase were examined using bacterially expressed protein. We show that retinol dehydratase can catalyze the transfer of the sulfonate moiety to small phenolic compounds and exhibits many functional similarities to the mammalian cytosolic sulfotransferases. The bisubstrate reaction that it catalyzes between retinol and the universal sulfonate donor 3'-phosphoadenosine 5' phosphosulfate seems to involve ternary complex formation and to proceed via a Random Bi Bi mechanism. In addition to the low nanomolar Km value for free retinol, retinol dehydratase is strongly inhibited by retinol metabolites, suggesting a preference for retinoids. Conversely, a number of tested mammalian cytosolic sulfotransferases do not utilize retinol, indicating that retinol is not a general substrate for sulfotransferases. PMID- 9857082 TI - Immunophilins: beyond immunosuppression. PMID- 9857083 TI - Nitrogen-carbon-linked (azolylphenyl)oxazolidinones with potent antibacterial activity against the fastidious gram-negative organisms Haemophilus influenzae and Moraxella catarrhalis. PMID- 9857085 TI - Discovery and structure-activity relationship of the first non-peptide competitive human glucagon receptor antagonists. AB - The first non-peptide competitive human glucagon receptor antagonist, 2 (benzimidazol-2-ylthio)-1-(3,4-dihydroxyphenyl)-1-ethan one, NNC 92-1687 (2), is described. This antagonist has a binding affinity of 20 microM (IC50) and a functional Ki = 9.1 microM at the human glucagon receptor. A structure-activity relationship (SAR) was obtained on this compound, and the results show that only the benzimidazole part can be changed without complete loss of affinity. Analogues with tert-butyl or benzyloxy groups in the 5-position of the benzimidazole moiety were found to be equipotent or slightly more potent, all displaying binding affinities around 5-20 microM. Most of the changes to the catechol and the linker gave compounds without any affinity toward the human glucagon receptor. The 3-hydroxy group could, however, in the presence of a 4 hydroxy group be changed to a methoxy or a chloro group while retaining affinity. PMID- 9857084 TI - A novel (benzodifuranyl)aminoalkane with extremely potent activity at the 5-HT2A receptor. PMID- 9857086 TI - Enantiospecific semisynthesis of (+)-almuheptolide-A, a novel natural heptolide inhibitor of the mammalian mitochondrial respiratory chain. AB - The development of novel styryl lactone derivatives as bioactive compounds and the semisynthesis of both 4,5-dialkoxylated eight-membered-ring lactones with a heptolide skeleton (almuheptolide-A (1) type) and 7-alkoxylated delta-lactones with a saturated furanopyrone skeleton (etharvensin (8) type) have been successfully achieved from the chiral unsaturated alpha-pyrone altholactone (7). This new method is a direct and one-step enantiospecific alkoxylation of altholactone (7) in concentrated acid medium, followed by formation of the eight membered-ring zeta-lactone. The reaction mechanism operating in the synthesis of the heptolide skeleton is postulated to be a direct Michael-type addition. Concerted opening of both the alpha-pyrone and tetrahydrofuran rings and subsequent intramolecular rearrangement with the ring closure lead to almuheptolide-A (1). This compound (1) and its diacetated derivative (1a) showed potent and selective inhibitory activity toward mammalian mitochondrial respiratory chain complex I. This mechanism of action, reported here for the first time, provides a possible explanation for the cytotoxic and antitumor activities previously described for related natural compounds. PMID- 9857087 TI - Side chain methyl substitution in the delta-opioid receptor antagonist TIPP has an important effect on the activity profile. AB - The delta-opioid antagonist H-Tyr-Tic-Phe-Phe-OH (TIPP-OH) or its C-terminal amide analogue was systematically modified topologically by substitution of each amino acid residue by all stereoisomers of the corresponding beta-methyl amino acid. The potency and selectivity (delta- vs mu- and kappa-opioid receptor) were evaluated by radioreceptor binding assays. Agonist or antagonist potency were assayed in the mouse vas deferens and in the guinea pig ileum. In the TIPP analogues containing L-beta-methyl amino acids the influence on delta-receptor affinity and on delta-antagonist potency is limited, the [(2S,3R)-beta MePhe3]TIPP-OH analogue being among the most potent delta-antagonists reported. In the D-beta-methyl amino acid series, the [D-beta-MeTic2] analogues are delta selective antagonists whereas [D-Tic2]TIPP-NH2 is a delta-agonist. NMR studies did not indicate any influence of the beta-methyl substituent on the conformation of the Tic residue. The [(2R,3S)-beta-MePhe3]TIPP-NH2 is a potent delta-agonist, its C-terminal carboxylic acid analogue being more delta-selective but displaying partial agonism in both the delta- and mu-bioassay. These results constitute further examples of a profound influence of beta-methyl substitution on the potency, selectivity, and signal transduction properties of a peptide. PMID- 9857088 TI - The bioactive conformation of aminoalkylindoles at the cannabinoid CB1 and CB2 receptors: insights gained from (E)- and (Z)-naphthylidene indenes. AB - The aminoalkylindoles (AAIs) are agonists at both the cannabinoid CB1 and CB2 receptors. To determine whether the s-trans or s-cis form of AAIs is their receptor-appropriate conformation, two pairs of rigid AAI analogues were studied. These rigid analogues are naphthylidene-substituted aminoalkylindenes that lack the carbonyl oxygen of the AAIs. Two pairs of (E)- and (Z)-naphthylidene indenes (C-2 H and C-2 Me) were considered. In each pair, the E geometric isomer is intended to mimic the s-trans form of the AAIs, while the Z geometric isomer is intended to mimic the s-cis form. Complete conformational analyses of two AAIs, pravadoline (2) and WIN-55, 212-2 (1), and of each indene were performed using the semiempirical method AM1. S-trans and s-cis conformations of 1 and 2 were identified. AM1 single-point energy calculations revealed that when 1 and each indene were overlayed at their corresponding indole/indene rings, the (E)- and (Z)-indenes were able to overlay naphthyl rings with the corresponding s-trans or s-cis conformer of 1 with an energy expense of 1.13/0.69 kcal/mol for the C-2 H (E/Z)-indenes and 0.82/0.74 kcal/mol for the C-2 Me (E/Z)-indenes. On the basis of the hypothesis that aromatic stacking is the predominant interaction of AAIs such as 1 at the CB receptors and on the demonstration that the C-2 H (E/Z)- and C-2 Me (E/Z)-indene isomers can mimic the positions of the aromatic systems in the s-trans and s-cis conformers of 1, the modeling results support the previously established use of indenes as rigid analogues of the AAIs. A synthesis of the naphthylidene indenes was developed using Horner-Wittig chemistry that afforded the Z isomer in the C-2 H series, which was not produced in significant amounts from an earlier reported indene/aldehyde condensation reaction. This approach was extended to the C-2 Me series as well. Photochemical interconversions in both the C-2 H and C-2 Me series were also successful in obtaining the less favored isomer. Thus, the photochemical process can be used to provide quantities of the minor isomers C-2 H/Z and C-2 Me/E. The CB1 and CB2 affinities as well as the activity of each compound in the twitch response of the guinea pig ileum (GPI) assay were assessed. The E isomer in each series was found to have the higher affinity for both the CB1 and CB2 receptors. In the rat brain membrane assay versus [3H]CP-55,940, the Ki's for the C-2 H/C-2 Me series were 2.72/2.89 nM (E isomer) and 148/1945 nM (Z isomer). In membrane assays versus [3H]SR141716A, a two-site model was indicated for the C-2 H/C-2 Me (E isomers) with Ki's of 10. 8/9.44 nM for the higher-affinity site and 611/602 nM for the lower-affinity site. For the Z isomers, a one-site model was indicated with Ki's of 928/2178 nM obtained for the C2 H/C-2 Me analogues, respectively. For the C-2 H/C-2 Me series, the CB2 Ki's obtained using a cloned cell line were 2.72/2.05 nM (E isomer) and 132/658 nM (Z isomer). In the GPI assay, the relative order of potency was C-2 H E > C-2 Me E > C-2 H Z > C-2 Me Z. The C-2 H E isomer was found to be equipotent with 1, while the C-2 Me Z isomer was inactive at concentrations up to 3.16 microM. Thus, results indicate that the E geometric isomer in each pair of analogues is the isomer with the higher CB1 and CB2 affinities and the higher pharmacological potency. Taken together, results reported here support the hypothesis that the s-trans conformation of AAIs such as 1 is the preferred conformation for interaction at both the CB1 and CB2 receptors and that aromatic stacking may be an important interaction for AAIs at these receptors. PMID- 9857089 TI - Identification of an opioid kappa receptor subtype-selective N-substituent for (+)-(3R,4R)-dimethyl-4-(3-hydroxyphenyl)piperidine. AB - A three-component library of compounds was prepared in parallel using multiple simultaneous solution-phase synthetic methodology. The compounds were biased toward opioid receptor antagonist activity by incorporating (+)-(3R,4R)-dimethyl 4-(3-hydroxyphenyl)piperidine (a potent, nonselective opioid pure antagonist) as one of the monomers. The other two monomers, which included N-substituted or unsubstituted Boc-protected amino acids and a range of substituted aryl carboxylic acids, were selected to add chemical diversity. Screening of these compounds in competitive binding experiments with the kappa opioid receptor selective ligand [3H]U69,593 led to the discovery of a novel kappa opioid receptor selective ligand, N-?(2'S)-[3-(4-hydroxyphenyl)propanamido]-3' methylbutyl?-(3R, 4R)-dimethyl-4-(3-hydroxyphenyl)piperidine (8, RTI-5989-29). Additional structure-activity relationship studies suggested that 8 possesses lipophilic and hydrogen-bonding sites that are important to its opioid receptor potency and selectivity. These sites appear to exist predominantly within the kappa receptor since the selectivity arises from a 530-fold loss of affinity of 8 for the mu receptor and an 18-fold increase in affinity for the kappa receptor relative to the mu-selective ligand, (+)-N-[trans-4-phenyl-2-butenyl]-(3R, 4R) dimethyl-4-(3-hydroxyphenyl)piperidine (5a). The degree of selectivity observed in the radioligand binding experiments was not observed in the functional assay. According to its ability to inhibit agonist stimulated binding of [35S]GTPgammaS at all three opioid receptors, compound 8 behaves as a mu/kappa opioid receptor pure antagonist with negligible affinity for the delta receptor. PMID- 9857090 TI - Biphenylsulfonamide endothelin antagonists: structure-activity relationships of a series of mono- and disubstituted analogues and pharmacology of the orally active endothelin antagonist 2'-amino-N- (3,4-dimethyl-5-isoxazolyl)-4'-(2 methylpropyl)[1, 1'-biphenyl]-2-sulfonamide (BMS-187308). AB - Substitution at the ortho position of N-(3,4-dimethyl-5-isoxazolyl) benzenesulfonamide led to the identification of the biphenylsulfonamides as a novel series of endothelin-A (ETA) selective antagonists. Appropriate substitutions on the pendant phenyl ring led to improved binding as well as functional activity. A hydrophobic group such as isobutyl or isopropoxyl was found to be optimal at the 4'-position. Introduction of an amino group at the 2' position also led to improved analogues. Combination of the optimal 4'-isobutyl substituent with the 2'-amino function afforded an analogue (20, BMS-187308) with improved ETA binding affinity and functional activity. Compound 20 also has good oral activity in inhibiting the pressor effect caused by an ET-1 infusion in rats. Doses of 10 and 30 micromol/kg iv 20 attenuated the pressor responses due to the administration of exogenous ET-1 to conscious monkeys, indicating that the compound inhibits the in vivo activity of endothelin-1 in nonhuman primates. PMID- 9857091 TI - Repaglinide and related hypoglycemic benzoic acid derivatives. AB - The structure-activity relationships in two series of hypoglycemic benzoic acid derivatives (5, 6) were investigated. Series 5 resulted from meglitinide (3) when the 2-methoxy was replaced by an alkyleneimino residue. Maximum activity was observed with the cis-3, 5-dimethyl-piperidino (5h) and the octamethyleneimino (5l) residues. Series 6 resulted from the meglitinide analogon 4 bearing an inversed amido function when the 2-methoxy, the 5-fluoro, and the alpha-methyl residue were replaced by a 2-piperidino, a 5-hydrogen, and a larger alpha-alkyl residue, respectively. An alkoxy residue ortho to the carboxy group further increased activity and duration of action in the rat. The most active racemic compound, 6al (R4 = isobutyl; R = ethoxy), turned out to be 12 times more active than the sulfonylurea (SU) glibenclamide (1). Activity was found to reside predominantly in the (S)-enantiomers. Compared with the SUs 1 and 2 (glimepiride), the most active enantiomer, (S)-6al (AG-EE 623 ZW; repaglinide; ED50 = 10 micro/kg po), is 25 and 18 times more active. Repaglinide turned out to be a useful therapeutic for type 2 diabetic patients; approval was granted recently by the FDA and the EMEA. From investigations on the pharmacophoric groups in compounds of type 5 and 6, it was concluded that in addition to the two already known-the acidic group (COOH; SO2NH) and the amidic spacer (CONH; NHCO) the ortho residue R1 (alkyleneimino; alkoxy; oxo) must be regarded as a third one. A general pharmacophore model suitable for hypoglycemic benzoic acid derivatives, SUs, and sulfonamides is proposed (Figure 6). Furthermore, from superpositions of low-energy conformations (LECs) of 1, 2, and (S)-6al, it was concluded that a common binding conformation (LEC II; Figure 10B) may exist and that differences in binding to the SU receptor and in the mechanism of insulin release between repaglinide and the two SUs may be due to specific hydrophobic differences. PMID- 9857092 TI - Resistance-modifying agents. 5. Synthesis and biological properties of quinazolinone inhibitors of the DNA repair enzyme poly(ADP-ribose) polymerase (PARP). AB - Clinical studies concerning the role of poly(ADP-ribose) polymerase (PARP) in the repair of drug- and radiation-induced DNA damage have been impeded by the poor solubility, lack of potency, and limited specificity of currently available inhibitors. A series of 2-alkyl- and 2-aryl-substituted 8-hydroxy-, 8-methoxy-, and 8-methylquinazolin-4(3H)-ones has been synthesized and evaluated for PARP inhibitory activity in permeabilized L1210 murine leukemia cells. 8-Methoxy- and 8-methylquinazolinones (14-34) were readily prepared by acylation of 3 substituted anthranilamides with the appropriate acid chloride, followed by base catalyzed cyclization. The requisite 8-hydroxyquinazolinones (6, 35-39) were synthesized by demethylation of the corresponding 8-methoxyquinazolinones with BBr3. N-Methylation of 8-methoxy-2-methylquinazolinone (15) with MeI, followed by O-demethylation by BBr3, afforded the control N3-methylquinazolinones 42 and 43, respectively. In general, an 8-hydroxy or 8-methyl substituent enhanced inhibitory activity in comparison with an 8-methoxy group. 2-Phenylquinazolinones were marginally less potent than the corresponding 2-methylquinazolinones, but the introduction of an electron-withdrawing or electron-donating 4'-substituent on the 2-aryl ring invariably increased potency. This was particularly evident in the 8-methylquinazolinone series (IC50 values 0.13-0.27 microM), which are among the most potent PARP inhibitors reported to date. N3-Methylquinazolinones 42 and 43 were essentially devoid of activity (IC50 values > 100 microM). In studies with L1210 cells in vitro, a concentration of 200 microM 8-hydroxy-2 methylquinazolinone (6, NU1025) (IC50 value 0.40 microM) potentiated the cytotoxicity of the monomethylating agent 5-(3-methyltriazen-1-yl)imidazole-4 carboxamide and gamma-radiation 3.5- and 1.4-fold, respectively, at the 10% survival level. PMID- 9857093 TI - (R)-(-)- and (S)-(+)-Synadenol: synthesis, absolute configuration, and enantioselectivity of antiviral effect. AB - Synthesis of (R)-(-)- and (S)-(+)-synadenol (1a and 2a, 95-96% ee) is described. Racemic synadenol (1a + 2a) was deaminated with adenosine deaminase to give (R)-( )-synadenol (1a) and (S)-(+)-hypoxanthine derivative 5. Acetylation of the latter compound gave acetate 6. Reaction with N, N-dimethylchloromethyleneammonium chloride led to 6-chloropurine derivative 7. Ammonolysis furnished (S)-(+) synadenol (2a). Absolute configuration of 1a was established by two methods: (i) synthesis from (R)-methylenecyclopropanecarboxylic acid (8) and (ii) X-ray diffraction of a single crystal of (-)-synadenol hydrochloride. Racemic methylenecyclopropanecarboxylic acid (10) was resolved by a modification of the described procedure. The R-enantiomer 8 was converted to ethyl ester 13 which was brominated to give vicinal dibromides 14. Reduction with diisobutylaluminum hydride then furnished alcohol 15 which was acetylated to the corresponding acetate 16. Alkylation-elimination procedure of adenine with 16 yielded acetates 17 and 18. Deprotection with ammonia afforded a mixture of Z- and E-isomers 1a and 19 of the R-configuration. Comparison with products 1a and 2a by chiral HPLC established the R-configuration of (-)-synadenol (1a). These results were confirmed by X-ray diffraction of a single crystal of (-)-synadenol hydrochloride. The latter forms a pseudosymmetric dimer with adenine-adenine base pairing in the lattice with the nucleobase in an anti-like conformation. Enantiomers 1a and 2a exhibit varied enantioselectivity toward different viruses. Both enantiomers are equipotent against human cytomegalovirus (HCMV) and varicella zoster virus (VZV). The S-enantiomer 2a is somewhat more effective than R-enantiomer 1a in herpes simplex virus 1 and 2 (HSV-1 and HSV-2) assays. By contrast, enantioselectivity of antiviral effect is reversed in Epstein-Barr virus (EBV) and human immunodeficiency virus type 1 (HIV-1) assays where the R enantiomer 1a is preferred. In these assays, the S-enantiomer 2a is less effective (EBV) or devoid of activity (HIV-1). PMID- 9857094 TI - Inactivation of O6-alkylguanine-DNA alkyltransferase. 1. Novel O6 (hetarylmethyl)guanines having basic rings in the side chain. AB - A number of novel guanine derivatives containing heterocyclic moieties at the O6 position have been synthesized using a purine quaternary salt which reacts with alkoxides under mild conditions. Initially O6-substituents were investigated in which the benzene ring of the known agent, O6-benzylguanine, was replaced by unsubstituted heterocyclic rings. The ability of these agents to inactivate the DNA repair protein O6-alkylguanine-DNA alkyltransferase (ATase), both as pure recombinant protein and in the human lymphoblastoid cell line Raji, has been compared with that of O6-benzylguanine. The present paper focuses on O6 substituents with basic rings, and under standard conditions several of them proved more effective than benzyl for inactivation of both recombinant and Raji ATase. Among the pyridine derivatives, the 2-picolyl compound 7 is not very active in contrast to the 3- and 4-picolyl compounds, and this influenced our choice of isomers of other basic ring systems for study. Since halogen substitution in the thiophene ring considerably increased the activity (17 versus 6), similar modifications in the pyridine series were examined. The more polar O6 substituents in this study are on the whole compatible with the stereochemical requirements of the ATase protein, and their pharmacological properties may be valuable in subsequent in vivo investigations, particularly the thenyl (6), 5 thiazolylmethyl (12), 5-bromothenyl (17), and 2-chloro-4-picolyl (21) derivatives. PMID- 9857095 TI - Prediction of binding affinities for TIBO inhibitors of HIV-1 reverse transcriptase using Monte Carlo simulations in a linear response method. AB - Monte Carlo (MC) simulations in combination with a linear response approach were used to estimate the free energies of binding for a series of 12 TIBO nonnucleoside inhibitors of HIV-1 reverse transcriptase. Separate correlations were made for the R6 and S6 absolute conformations of the inhibitors, as well as for the analogous N6-monoprotonated species. Models based upon the neutral unbound inhibitors produced overall better fits to experimental values than did those using the protonated unbound inhibitors, with only slight differences between the neutral R6 and S6 cases. The best results were obtained with a three parameter linear response equation containing van der Waals (alpha), electrostatic (beta), and solvent accessible surface area (SASA, gamma) terms. The averaged (R6 and S6) rms error was approximately 0.88 kcal/mol for the observed range of 4.06 kcal/mol in inhibitor activities. The averaged values of alpha, beta, and gamma were -0.150, 0.114, and 0. 0286, respectively. Omission of the alpha term gave beta 0.152 and gamma 0.022 with a rms of 0.92. The unweighted van der Waals components were found to be highly attractive but failed to correlate well across the series of inhibitors. Contrastingly, while the electrostatic components are all repulsive, they show a direct correlation with inhibitor activity as measured by DeltaGbinding. The role of gamma is primarily to produce an overall negative binding energy, and it can effectively be replaced with a negative constant. During the MC simulations of the unbound solvated inhibitors, the R6 and S6 absolute conformations do not interconvert due to the formation of a favorable hydrogen bond to solvent. In the complex, however, interconversion of these conformations of the inhibitor is observed during the course of the simulations, a phenomenon which is apparently not observed in the crystalline state of the complex. Hydrogen bonding of the inhibitor to the backbone NH of K101 and the lack of such an interaction with the C=O of K101 or with solvent correlate with enhanced activity, as does the ability to assume a number of different orientations of the inhibitor dimethylallyl moiety with respect to residues Y181 and Y188 while retaining contact with W229. Overall, the use of a combination of MC simulation with a linear response method shows promise as a relatively rapid means of estimating inhibitor activities. This approach should be useful in the preliminary evaluation of potential modifications to known inhibitors to enhance activity. PMID- 9857096 TI - Chemoprotective properties of phenylpropenoids, bis(benzylidene)cycloalkanones, and related Michael reaction acceptors: correlation of potencies as phase 2 enzyme inducers and radical scavengers. AB - Induction of phase 2 enzymes (e.g., glutathione transferases, NAD(P)H:quinone reductase, glucuronosyltransferases, epoxide hydrolase) is a major strategy for reducing the susceptibility of animal cells to neoplasia and other forms of electrophile toxicity. In a search for new chemoprotective enzyme inducers, a structure-activity analysis was carried out on two types of naturally occurring and synthetic substituted phenylpropenoids: (a) Ar-CH=CH-CO-R, where R is OH, OCH3, CH3, or Ar, including cinnamic, coumaric, ferulic, and sinapic acid derivatives, their ketone analogues, and chalcones; and (b) bis(benzylidene)cycloalkanones, Ar-CH=C(CH2)n(CO)C=CH-Ar, where n = 5, 6, or 7. The potencies of these compounds in inducing NAD(P)H:quinone reductase activity in murine hepatoma cells paralleled their Michael reaction acceptor activity (Talalay, P.; De Long, M. J.; Prochaska, H. J. Proc. Natl. Acad. Sci. U.S.A. 85, 1988, 8261-8265). Unexpectedly, the bis(benzylidene)cycloalkanones also powerfully quenched the lucigenin-derived chemiluminescence evoked by superoxide radicals. Introduction of o-hydroxyl groups on the aromatic rings of these phenylpropenoids dramatically enhanced their potencies not only as inducers for quinone reductase but also as quenchers of superoxide. These potentiating o hydroxyl groups are hydrogen-bonded, as shown by moderate downfield shift of their proton NMR resonances and their sensitivities to the solvent environment. The finding that the potencies of a series of bis(benzylidene)cycloalkanones in inducing quinone reductase appear to be correlated with their ability to quench superoxide radicals suggests that the regulation of phase 2 enzymes may involve both Michael reaction reactivity and radical quenching mechanisms. PMID- 9857097 TI - Self-immolative nitrogen mustard prodrugs for suicide gene therapy. AB - Four new potential self-immolative prodrugs derived from phenol and aniline nitrogen mustards, four model compounds derived from their corresponding fluoroethyl analogues and two new self-immolative linkers were designed and synthesized for use in the suicide gene therapy termed GDEPT (gene-directed enzyme prodrug therapy). The self-immolative prodrugs were designed to be activated by the enzyme carboxypeptidase G2 (CPG2) releasing an active drug by a 1, 6-elimination mechanism via an unstable intermediate. Thus, N-[(4-?[4-(bis?2 chloroethyl?amino)phenoxycarbonyloxy]methyl?pheny l)c arbamoyl]-L-glutamic acid (23), N-[(4-?[4-(bis?2-chloroethyl?amino)phenoxycarbonyloxy]methyl?pheno xy) carbonyl]-L-glutamic acid (30), N-[(4-?[N-(4-?bis[2 chloroethyl]amino?phenyl)carbamoyloxy]methyl?+ ++phen oxy)carbonyl]-L-glutamic acid (37), and N-[(4-?[N-(4-?bis[2-chloroethyl]amino?phenyl)carbamoyloxy]methyl?+ ++phen yl)carbamoyl]-L-glutamic acid (40) were synthesized. They are bifunctional alkylating agents in which the activating effects of the phenolic hydroxyl or amino functions are masked through an oxycarbonyl or a carbamoyl bond to a benzylic spacer which is itself linked to a glutamic acid by an oxycarbonyl or a carbamoyl bond. The corresponding fluoroethyl compounds 25, 32, 42, and 44 were also synthesized. The rationale was to obtain model compounds with greatly reduced alkylating abilities that would be much less reactive with nucleophiles compared to the corresponding chloroethyl derivatives. This enabled studies of these model compounds as substrates for CPG2, without incurring the rapid and complicated decomposition pathways of the chloroethyl derivatives. The prodrugs were designed to be activated to their corresponding phenol and aniline nitrogen mustard drugs by CPG2 for use in GDEPT. The synthesis of the analogous novel parent drugs (21b, 51) is also described. A colorectal cell line was engineered to express CPG2 tethered to the outer cell surface. The phenylenediamine compounds were found to behave as prodrugs, yielding IC50 prodrug/IC50 drug ratios between 20- and 33-fold (for 37 and 40) and differentials of 12-14-fold between CPG2-expressing and control LacZ-expressing clones. The drugs released are up to 70-fold more potent than 4-[(2-chloroethyl)(2 mesyloxyethyl)amino]benzoic acid that results from the prodrug 4-[(2 chloroethyl)(2-mesyloxyethyl)amino]benzoyl-L-glutamic acid (CMDA) which has been used previously for GDEPT. These data demonstrate the viability of this strategy and indicate that self-immolative prodrugs can be synthesized to release potent mustard drugs selectively by cells expressing CPG2 tethered to the cell surface in GDEPT. PMID- 9857098 TI - Synthesis and potent antifolate activity and cytotoxicity of B-ring deaza analogues of the nonpolyglutamatable dihydrofolate reductase inhibitor Nalpha-(4 amino-4-deoxypteroyl)-Ndelta-hemiphthaloyl- L-ornithine (PT523). AB - Six new B-ring analogues of the nonpolyglutamatable antifolate Nalpha-(4-amino-4 deoxypteroyl)-Ndelta-hemiphthaloy l-L-ornithine (PT523, 3) were synthesized with a view to determining the effect of modifications at the 5- and/or 8-position on dihydrofolate reductase (DHFR) binding and tumor cell growth inhibition. The 5- and 8-deaza analogues were prepared from methyl 2-L-amino-5-phthalimidopentanoate and 4-amino-4-deoxy-N10-formyl-5-deaza- and -8-deazapteroic acid, respectively. The 5,8-dideaza analogues were prepared from methyl 2-L-[(4-aminobenzoyl)amino]-5 phthalimidopentanoate and 2, 4-diaminoquinazoline-6-carbonitriles. The Ki for inhibition of human DHFR by the 5-deaza and 5-methyl-5-deaza analogues was about the same as that of 3 (0.35 pM), 11-fold lower than that of aminopterin (AMT, 1), and 15-fold lower than that of methotrexate (MTX, 2). However the Ki of the 8 deaza analogue was 27-fold lower than that of 1, and that of the 5,8-dideaza, 5 methyl-5,8-dideaza, and 5-chloro-5,8-dideaza analogues was approximately 50-fold lower. This trend was consistent with the published literature on the corresponding DHFR inhibitors with a glutamate side chain. In colony formation assays against the human head and neck squamous carcinoma cell line SCC25 after 72 h of treatment, the 5- and 8-deaza analogues were approximately as potent as 3, whereas the 5,8-dideaza analogue was 3 times more potent. 5-Methyl and 5 chloro substitution was also favorable, with the 5-methyl-5-deaza analogue being 2. 5-fold more potent than the 5-deaza analogue. However the effect of 5-methyl substitution was less pronounced in the 5,8-dideaza analogues than in the 5-deaza analogues. The 5-chloro-5,8-dideaza analogue of 3 was the most active member of the series, with an IC50 = 0.33 nM versus 1.8 nM for 3 and 15 nM for MTX. The 5 methyl-5-deaza analogue of 3 was also tested at the National Cancer Institute against a panel of 50 human tumor cell lines in culture and was consistently more potent than 3, with IC50 values in the low-nanomolar to subnanomolar range against most of the tumors. Leukemia and colorectal carcinoma cell lines were generally most sensitive, though good activity was also observed against CNS tumors and carcinomas of the breast and prostate. The results of this study demonstrate that B-ring analogues of 3 inhibit DHFR activity and tumor cell colony formation as well as, or better than, the parent compound. In view of the fact that 3 and its B-ring analogues cannot form polyglutamates, their high cytotoxicity relative to the corresponding B-ring analogues of AMT is noteworthy. PMID- 9857100 TI - Vasoconstrictor responsiveness of portal hypertensive vessels. PMID- 9857099 TI - Design and synthesis of novel alpha1a adrenoceptor-selective dihydropyridine antagonists for the treatment of benign prostatic hyperplasia. AB - We report the synthesis and evaluation of novel alpha1a adrenoceptor subtype selective antagonists. Systematic modification of the lipophilic 4,4 diphenylpiperidinyl moiety of the dihydropyridine derivatives 1 and 2 provided several highly selective and potent alpha1a antagonists. From this series, we identified the 4-(methoxycarbonyl)-4-phenylpiperidine analogue SNAP 5540 (-) [(-) 63] for further characterization. When examined in an isolated human prostate tissue assay, this compound was found to have a Ki of 2.8 nM, in agreement with the cloned human receptor binding data (Ki = 2.42 nM). Further evaluation of the compound in isolated dog prostate tissue showed a Ki of 3.6 nM and confirmed it to be a potent antagonist (Kb = 1.6 nM). In vivo, this compound effectively blocked the phenylephrine-stimulated increase in intraurethral pressure (IUP) in mongrel dogs, at doses which did not significantly affect the arterial pressure (diastolic blood pressure, DBP), with a DBP Kb/IUP Kb ratio of 16. In addition, ( )-63 also showed greater than 40 000-fold selectivity over the rat L-type calcium channel and 200-fold selectivity over several G protein-coupled receptors, including histamine and serotonin subtypes. These findings prove that alpha1a adrenoceptor-subtype selective antagonists such as (-)-63 may be developed as uroselective agents for an improved treatment of BPH over nonselective alpha1 antagonists such as prazosin and terazosin, with fewer side effects. PMID- 9857101 TI - Renal effects of brain natriuretic peptide in patients with congestive heart failure. AB - The effect of a continuous infusion of human brain natriuretic peptide, 2 pmol.min-1.kg-1, during 60 min was studied in nine patients with congestive heart failure and in 10 healthy control subjects. Brain natriuretic peptide increased from 1.6 to 101 pmol/l in control subjects and from 25 to 173 pmol/l in congestive heart failure during infusion. Urinary sodium excretion increased significantly in both congestive heart failure (60%) and control subjects (71%), but the absolute increase was significantly lower in congestive heart failure (27 micromol/min) than in control subjects (190 micromol/min). Urinary flow rate did not change. The lithium clearance technique was used to evaluate the segmental tubular function; the distal fractional reabsorption of sodium decreased significantly less in congestive heart failure (DFRNa: -0.8%) than in control subjects (DFRNa: -3.7%). Baseline values for glomerular filtration rate and renal plasma flow were reduced in congestive heart failure, but brain natriuretic peptide induced no significant changes between congestive heart failure and control subjects. Brain natriuretic peptide induced the same absolute increase in secondary messenger cGMP in plasma and urine in both patients and healthy subjects. It is concluded that the natriuretic response to brain natriuretic peptide infusion was impaired in patients with congestive heart failure compared with healthy subjects, and it is likely that the impaired natriuretic response was caused by a reduced responsiveness in the distal part of the nephron. PMID- 9857102 TI - Angiotensin-converting enzyme inhibition restores the diffusing capacity for carbon monoxide in patients with chronic heart failure by improving the molecular diffusion across the alveolar capillary membrane. AB - Conductance of alveolar capillary membrane (DM) and capillary blood volume (VC) are the subcomponents of the pulmonary diffusing capacity for carbon monoxide (DLCO). In chronic heart failure, stress failure of the membrane provides a mechanism for reduced DM and subsequent impairment of DLCO. Angiotensin converting enzyme inhibition improves DLCO in patients with chronic heart failure. This study was aimed at investigating which of the two subcomponents of DLCO is affected by angiotensin-converting enzyme inhibitors. Twenty-seven patients with NYHA class II to III chronic heart failure (group 1) and 13 age- and sex-matched normal subjects underwent pulmonary function testing with determination of DM and VC, while receiving placebo and 48 h and 1 and 2 months after starting enalapril treatment (10 mg twice daily). Nine similar patients (group 2) received isosorbide dinitrate (40 mg thrice daily) for a month then enalapril for another month, and underwent pulmonary function testing at 48 h and 1 month after starting treatments. Effects of angiotensin-converting enzyme inhibition in normal controls were not significant in the short- or mid-term. In group 1 patients, the only change observed at 48 h was a reduction in VC (probably due to a decrease in capillary pulmonary pressure). There was a marked increase in DM to a similar extent at 1 and 2 months, resulting in a significant improvement in DLCO despite a decrease in VC. In group 2 patients, nitrates failed to improve DLCO and DM, whereas enalapril was as effective as in group 1. These observations suggest a modulatory effect of angiotensin-converting enzyme inhibition on the membrane function which emerges gradually and persists over time and is probably dissociated from changes in pulmonary capillary pressure and VC. Chronic heart failure disturbs the alveolar capillary interface and increases gas diffusion resistance; angiotensin-converting enzyme inhibition restores the diffusive properties of the membrane and gas transfer, and protects the lung when the heart is failing. PMID- 9857103 TI - Impaired sympathetic response before intradialytic hypotension: a study based on spectral analysis of heart rate and pressure variability. AB - The purpose of this study was to evaluate the autonomic response to standard haemodialysis and the changes associated with the onset of intradialytic hypotension in 12 normotensive patients with uraemia. Power spectra of R-R interval and of blood pressure fluctuations were obtained during a standard dialysis session and estimated in the low-frequency (LF, 30-150 mHz) and high frequency (HF, 150-400 mHz) range. The absolute power of the LF component of blood pressure variations and the LF/HF ratio of R-R interval were assumed as indexes of sympathetic activity. Standard haemodialysis induced hypotension in six patients (unstable) while a minor pressure decline was present in the other six (stable). Normalized blood volume before dialysis and percentage volume reduction were similar in the two groups. Tachycardia in response to pressure and volume decrease was more pronounced in stable than in unstable patients, as evidenced by a higher slope of the relation between R-R interval and systolic blood pressure (7.9 versus 0.9 ms/mmHg, P<0.01). Sympathetic tone was enhanced during early dialysis in all patients (+2+/-1 for R-R LF/HF ratio, +2.4+/-0.6 mmHg2 and +7.2+/-2 mmHg2 for absolute LF power of diastolic and of systolic blood pressure respectively, P<0.05), compared with baseline predialysis values. During late dialysis, unstable patients showed an impairment of sympathetic activation which preceded hypotension and was maximal during the crisis (-2.9+/-1.4 for R-R LF/HF ratio, -2.7+/-1.4 mmHg2 and -8.6+/-4.0 mmHg2 for absolute LF power of diastolic and of systolic blood pressure respectively, P<0.05). On the contrary, stable patients showed constantly elevated indexes (+3.7+/-1.4 for R-R LF/HF ratio, +5.9+/-2.7 mmHg2 and +13.3+/-6.2 mmHg2 for LF of diastolic and of systolic blood pressure, P<0.05). Values returned to predialysis levels after the end of the dialysis session in all patients. We conclude that standard haemodialysis activates a marked and reversible sympathetic response in both stable and unstable uraemic patients. However, in unstable patients, such activation is impaired in late dialysis, therefore contributing to the onset of the hypotensive crisis. PMID- 9857104 TI - Increased plasma levels of adrenomedullin in patients with pulmonary hypertension. AB - Adrenomedullin, a potent hypotensive peptide, reduces blood pressure and pulmonary vascular resistance, and increases pulmonary blood flow. The mRNA for adrenomedullin and its receptor is highly expressed in the lung, suggesting a regulatory role for adrenomedullin in the pulmonary circulation. To investigate the clinical significance of adrenomedullin in patients with pulmonary hypertension, we studied the relationship between plasma levels of adrenomedullin and pulmonary haemodynamics. Venous, arterial and pulmonary arterial blood samples were obtained during cardiac catheterization and plasma levels of adrenomedullin were measured by specific radioimmunoassay in 33 consecutive patients with severe pulmonary hypertension (12 cases of primary pulmonary hypertension, 21 with chronic thromboembolic pulmonary hypertension; age 49+/-16 years, mean pulmonary arterial pressure 50+/-15mmHg). In addition, plasma levels of adrenomedullin were measured before and after acute nitric oxide inhalation. The changes in plasma adrenomedullin during the follow-up period of 10.3+/-4.3 months were also evaluated (n=5). Sixty-two healthy subjects served as the control group. Adrenomedullin was measured in an antecubital vein in the controls. Plasma levels of adrenomedullin were significantly higher in the patients with pulmonary hypertension than in the control subjects (10.1+/-8.7 versus 4.9+/-1.1pmol/l, P<0.01). Plasma levels of adrenomedullin, expressed as their natural logarithm, were significantly correlated with mean right atrial pressure (r=0.71, P<0.01), stroke volume (r=-0.63, P<0.01), total pulmonary resistance (r=0.60, P<0.01), mean pulmonary arterial pressure (r=0.37, P<0.05), and the natural logarithm of plasma atrial natriuretic peptide (r=0. 63, P<0.01). Plasma levels of adrenomedullin did not change significantly after nitric oxide inhalation, but significantly increased in association with the elevation of the total pulmonary resistance during the long-term follow-up period. These results suggest that plasma levels of adrenomedullin increase in proportion to the extent of pulmonary hypertension. PMID- 9857105 TI - Portal hypertension increases vasoconstrictor responsiveness of rat aorta. AB - We have examined the effects of pre-hepatic portal hypertension on the responsiveness of aorta from Wistar and Sprague-Dawley rats. Rats were made portal hypertensive by creating a calibrated portal vein stenosis, or sham operated. In rat aorta, there was no significant difference between portal hypertensive and sham-operated animals in the contractile potency of KCl, noradrenaline or phenylephrine. In aortas from Wistar rats, the maximum response to KCl (0.71+/-0.12 g) and noradrenaline (1.00+/-0.17 g) but not phenylephrine (0.86+/-0.10 g) in portal hypertensive animals was significantly increased compared with that in sham-operated animals (0.45+/-0.04 g, 0.57+/-0.07 g, 0.71+/ 0.05 g respectively). In aortas from Sprague-Dawley rats, the maximum response to KCl (1. 21+/-0.21 g) and phenylephrine (1.54+/-0.30 g) but not noradrenaline (0.93+/-0.09 g) in portal hypertensive animals was significantly increased compared with that in sham-operated animals (0.59+/-0.09 g, 0.76+/-0.11 g, 1.04+/ 0.10 g respectively). There was no difference between portal hypertensive and sham-operated Wistar rats in the affinity or maximum number of binding sites for [3H]prazosin to alpha1-adrenoceptors in cardiac ventricular membranes. It is concluded that portal hypertension tends to produce an increase rather than a decrease in the contractile response to vasoconstrictors in aorta from both Wistar and Sprague-Dawley rats. This suggests that the diminished responsiveness to vasoconstrictors reported in portal hypertensive rats in vivo is not due to a diminished responsiveness at the level of the vascular smooth muscle. PMID- 9857106 TI - Autonomic nervous system dysfunction in sclerodermic and primary Raynaud's phenomenon. AB - Our aim was to investigate the sympathetic hyperactivity of systemic sclerosis that may lead to greater morbidity and mortality from cardiovascular events. We analysed the sympathetic (low-frequency) and vagal (high-frequency) components of heart rate variability, in supine and upright positions, in 10 patients with systemic sclerosis, 12 patients with primary Raynaud's phenomenon and 14 controls. We also analysed lung function in order to evaluate a possible link between heart rate variability and ventilation parameters. Heart rate variability was reduced in the supine position in subjects with systemic sclerosis both in comparison with primary Raynaud's phenomenon (total power: 1103+/-156 versus 3302+/-486 ms2, P<0.004) and control subjects (3148+/-422 ms2, P<0.002). Low frequency power was higher in patients with systemic sclerosis than in the controls (54.5+/-4.5 versus 42.5+/-3.5 normalized units, P<0.01). During tilt, the change in heart rate was +44% in controls, +24% in subjects with primary Raynaud's phenomenon, and only +17% in the patients with systemic sclerosis (P<0.01 versus controls). In patients with systemic sclerosis we found a significant correlation between high-frequency power and the indices of lung function (residual volume: r2=0.5143, P<0.01; total lung capacity: r2=0.5142, P<0.01, vital capacity: r2=0.3789, P<0.05). Heart rate variability was reduced and sympathetic output increased in patients with systemic sclerosis. Subjects with primary Raynaud's phenomenon were characterized by normal heart rate variability and by some degree of sympathetic hyperactivity. During tilting, subjects with systemic sclerosis maintained an unmodified heart rate variability, thus suggesting an impaired baroceptor modulation of the autonomic control. The negative correlation between high-frequency power and indices of respiratory insufficiency in patients with systemic sclerosis suggests that the pulmonary structure plays an important role in the modulation of heart rate variability. PMID- 9857107 TI - Comparative effects of dilator drugs on human penile dorsal artery and deep dorsal vein. AB - The present study was designed to characterize the response of human penile dorsal artery and deep dorsal vein to dilator drugs used in the diagnosis and treatment of erectile dysfunction with special emphasis on the effects on sympathetic neurotransmission. Ring segments of penile dorsal artery and deep dorsal vein were obtained from 20 multi-organ donors during procurement of organs for transplantation. The rings (3 mm long) were suspended in organ bath chambers for isometric recording of tension. We then studied the relaxant responses to prostaglandin E1 (PGE1), vasoactive intestinal peptide (VIP), papaverine (PV), sodium nitroprusside (SNP) and linsidomine chlorhydrate (SIN-1), and analysed the effects of these drugs on contractions induced by stimulation of perivascular sympathetic nerves. In artery and vein rings contracted by noradrenaline, all the drugs tested caused concentration-dependent relaxation. The order of potencies in terms of IC50 values (concentration of agonist causing 50% of the maximal relaxation) was PGE1=VIP>SNP>SIN-1=PV. Both arteries and veins contracted to electrical field stimulation (15 V, 0.5-2 Hz, 0.2 ms duration for 15 s) in a frequency-dependent manner. All relaxant drugs caused concentration-dependent inhibition of neurogenic contractions; the relative order of potencies was PGE1>VIP>SNP>SIN-1=PV. It is concluded that inhibition of sympathetic activity constitutes an effective relaxing mechanism in penile dorsal artery and vein. Modulation of sympathetic activity together with the direct effects on smooth muscle should be considered to evaluate adequately the efficacy of relaxant drugs to increase human penile blood supply. PMID- 9857108 TI - Increased excretion of nitric oxide in exhaled air of patients with chronic renal failure. AB - Nitric oxide exerts multiple effects on renal function. It remains unclear whether endogenous nitric oxide production is increased or decreased in patients with chronic renal failure. To evaluate endogenous nitric oxide production in these patients we studied exhaled nitric oxide output by an ozone chemiluminescence method and plasma NO2(-)/NO3(-) levels by the Griess method in 40 patients with end-stage chronic renal failure who underwent regular continuous ambulatory peritoneal dialysis (n=30) or haemodialysis (n=10), and in 28 healthy subjects. Patients with chronic renal failure had a higher exhaled nitric oxide concentration [39+/-3 versus 19+/-1 parts per billion, (mean+/-S.E.M.), P<0.0001], a greater nitric oxide output (177+/-11 versus 96+/-7 nl.min-1.m-2, P<0.001) and a higher plasma NO2(-)/NO3(-) concentration (96+/-14 versus 33+/-4 micromol, P<0.01) than controls. These values did not differ between patients on haemodialysis and those on continuous ambulatory peritoneal dialysis. Patients with chronic renal failure had significantly higher plasma concentrations of both interleukin-1beta and interferon-gamma than controls. The exhaled nitric oxide output did not correlate with plasma NO2(-)/NO3(-) or with peritoneal dialysate NO2(-)/NO3(-), but plasma NO2(-)/NO3(-) correlated with dialysate NO2(-)/NO3(-) in patients who underwent continuous ambulatory peritoneal dialysis (r=0.77, P<0.01). Haemodialysis for 4 h acutely decreased plasma NO2(-)/NO3(-) (92+/-17 versus 50+/-8 micromol, P<0.05) and cGMP concentration (16.5+/-4.3 versus 5.1+/ 1. 7 pmol/ml, P<0.01), but did not decrease exhaled nitric oxide output. The increase in exhaled nitric oxide with the simultaneous increase in circulating cytokines suggests that nitric oxide synthase seems to be induced significantly in patients with chronic renal failure. Increased endogenous nitric oxide production may have a pathophysiological role in patients with uraemia. PMID- 9857109 TI - Alpha-lipoic acid reduces expression of vascular cell adhesion molecule-1 and endothelial adhesion of human monocytes after stimulation with advanced glycation end products. AB - Advanced glycation end products (AGEs) have been identified as relevant mediators of late diabetic complications such as atherosclerotic disease. The endothelial migration of monocytes is one of the first steps in atherogenesis and monocyte endothelial interaction itself is linked to the expression of adhesion molecules like vascular cell adhesion molecule-1 (VCAM-1). Recently, stimulation of VCAM-1 by AGEs has been demonstrated. Since endothelial stimulation by AGEs is followed by generation of oxygen free radicals with subsequent activation of nuclear transcription factor kappaB, we investigated the influence of alpha-lipoic acid on the expression of VCAM-1 and monocyte adherence to endothelial cells in vitro by means of cell-associated chemiluminescence assays and quantitative reverse transcriptase polymerase chain reaction using a constructed recombinant RNA standard. We found that alpha-lipoic acid was able to decrease the number of VCAM 1 transcripts from 41. 0+/-11.2 to 9.5+/-4.7 RNA copies per cell in AGE stimulated cell cultures. Furthermore, expression of VCAM-1 was suppressed in a time- and dose-dependent manner by alpha-lipoic acid as shown by chemiluminescence endothelial cell assay. Pretreatment of endothelial cells with 0.5 mM or 5 mM alpha-lipoic acid reduced AGE-induced endothelial binding of monocytes from 22.5+/-2.9% to 18. 3+/-1.9% and 13.8+/-1.8% respectively. Thus, we suggest that extracellularly administered alpha-lipoic acid reduces AGE-albumin induced endothelial expression of VCAM-1 and monocyte binding to endothelium in vitro. These in vitro results may contribute to the understanding of a potential antioxidative treatment of atherosclerosis. PMID- 9857110 TI - Relationship of serum levels of interleukin-6, soluble interleukin-6 receptor and tumour necrosis factor receptors to the acute-phase protein response in advanced pancreatic cancer. AB - The level of the acute-phase response is a major predictor of survival in patients with advanced pancreatic cancer. This study examines the association between the acute-phase protein response, as determined by serum C-reactive protein, and serum levels of interleukin-6, soluble interleukin-6 receptor and the soluble tumour necrosis factor receptors in patients with pancreatic cancer. Thirty-four blood samples were collected from 13 patients with advanced pancreatic cancer. Samples were also collected from six healthy subjects. Levels of C-reactive protein, interleukin-6, soluble interleukin-6 receptor and soluble tumour necrosis factor receptors 55 and 75 were measured by indirect ELISA. Serum levels of C-reactive protein, interleukin-6 and soluble tumour necrosis factor receptors 55 and 75 were significantly higher in cancer patients than in controls. Levels of serum soluble interleukin-6 receptor were not significantly different between the two groups. In cancer patients, a significant positive association was found between the level of the acute-phase protein response and serum levels of interleukin-6, soluble tumour necrosis factor receptor 55 and soluble tumour necrosis factor receptor 75. No association was found between levels of soluble interleukin-6 receptor and any other factor. There is no significant relationship between the level of soluble interleukin-6 receptor and the acute-phase protein response in vivo and the biological role of soluble interleukin-6 receptor in the chronic inflammatory component of cachexia remains unclear. PMID- 9857111 TI - Macrophage-mediated lysis of a beta-cell line, tumour necrosis factor-alpha release from bacillus Calmette-Guerin (BCG)-activated murine macrophages and interleukin-8 release from human monocytes are dependent on extracellular glutamine concentration and glutamine metabolism. AB - Macrophages and monocytes are cells with a large capacity for cytokine production. Cytokines produced by these cells are not preformed and released upon stimulation, but must be transcribed and translated. Although much is known concerning the regulation of the latter processes at the molecular level, the role of exogenous amino acids in the secretory process has not been actively investigated. Glutamine is utilized by macrophages at a much faster rate than any other amino acid. The role for high rates of glutamine utilization in macrophages or monocytes is not fully understood. We demonstrate here that the rates of lipopolysaccharide-stimulated tumour necrosis factor-alpha secretion from bacillus Calmette-Guerin (BCG)-activated murine peritoneal macrophages and lipopolysaccharide-stimulated interleukin-8 production from human monocytes are dependent upon extracellular glutamine concentration. We also demonstrate that potent inhibition of cytokine production can be achieved by incubating macrophages or monocytes in the presence of the glutaminase inhibitor 6-diazo-5 oxo-norleucine. On co-culture of BCG-activated macrophages and the clonal pancreatic beta-cell line BRIN-BD11, macrophage-specific beta-cell death was significantly reduced on prior exposure of macrophages to 6-diazo-5-oxo norleucine. Thus glutamine metabolism may be essential for generation of cytotoxic products from macrophages, including tumour necrosis factor-alpha. PMID- 9857112 TI - Inhibition of mitogen-activated proliferation of human peripheral lymphocytes in vitro by propionic acid. AB - Recurrent infections are common features in patients affected by propionic acidaemia (McKusick 232000) and methylmalonic acidaemia (McKusick 251000). Since these disorders are biochemically characterized by tissue accumulation of propionic acid and methylmalonic acid respectively, it is possible that these compounds may act as immunosuppressants. We therefore investigated the effect of propionate and methylmalonate on cellular growth of human peripheral lymphocytes stimulated in vitro by phytohaemagglutinin, concanavalin A and pokeweed mitogen, a recognized test of cellular immunocompetence. Lymphocytes were cultured in flat bottomed 96-well microplates at 37 degrees C for 96 h (phytohaemagglutinin and concanavalin A) or 144 h (pokeweed mitogen) in the presence of one mitogen at different concentrations and of one acid added at doses of 1.0, 2.5 or 5.0 mM. Cell blastogenesis was measured by the incorporation of tritiated thymidine into cellular DNA and compared with that of identical cultures with no acid added (controls). A consistent and progressive inhibitory effect of propionic acid with increasing concentrations in culture was identified with all mitogens and was more pronounced with pokeweed mitogen. Lymphocyte blastogenesis was not altered in the presence of methylmalonic acid. The effect of propionate was observed only when the drug was added at the beginning (phytohaemagglutinin-activated) or until 24 h (concanavalin A- and pokeweed mitogen-activated) of culture. The viability of lymphocytes after treatment with the drug, as assessed by the Trypan Blue exclusion test, revealed no change when compared with the same untreated lymphocytes, indicating no lymphocytotoxic activity. In conclusion, propionic acid, which accumulates in tissues of patients with propionic acidaemia, causes 'in vitro' immunosuppression, which may be related to the recurrent infections characteristic of these patients. PMID- 9857113 TI - Changes in indices of antioxidant status, lipid peroxidation and inflammation in human skeletal muscle after eccentric muscle actions. AB - This study investigated the effects of chronic muscle inflammation on indices of antioxidant status and muscle injury after eccentric exercise. Eight subjects each performed 70 maximal voluntary eccentric muscle actions on an isokinetic dynamometer, using the knee extensors of a single leg. Venous blood samples were collected into serum and EDTA tubes 5 and 3 days before exercise, immediately before exercise, and then again on days 3, 4, 5, 6, 7, 10 and 12 after the bout. Needle biopsies were taken from the vastus lateralis of six subjects, a week before exercise (baseline), and again on days 4 and 7 post-exercise. The concentrations of malondialdehyde in plasma and muscle were used as markers of lipid peroxidation. Creatine kinase activity, beta-glucuronidase activity and total antioxidant capacity were determined in serum. In muscle, aqueous and bound total antioxidant capacity, the aqueous sulphydryl concentration, and beta glucuronidase and glucose-6-phosphate dehydrogenase activity were determined. No changes were detected in serum total antioxidant capacity, serum creatine kinase and beta-glucuronidase after the baseline biopsy. After exercise serum creatine kinase and beta-glucuronidase were elevated although other serum measures were unchanged. In muscle, aqueous and bound total antioxidant capacity, sulphydryls, glucose-6-phosphate dehydrogenase and beta-glucuronidase were all elevated. Despite evidence of inflammation in this study, muscle antioxidant status was not compromised, and malondialdehyde was unaltered in muscle and plasma. Therefore, this study provides no evidence that chronic muscle inflammation compromises antioxidant status or increases lipid peroxidation. PMID- 9857114 TI - Chronic fatigue syndrome: a hypothesis focusing on the autonomic nervous system. AB - Chronic fatigue syndrome is a debilitating illness of unknown aetiology, with estimated levels of prevalence of up to about 8. 7/100 000 in the U.S.A. Like pain fatigue it is a personal, emotionally rich experience, which may originate from peripheral or central sites (or both). The nature of the symptoms is complex and reflects the interaction of the patient with the environment and cultural milieu. Accordingly the common use of the same terminology for different types of fatigue may be misleading. Autonomic activation is a key component of both real and simulated physical exercise. Alterations in autonomic nervous system activity are a key component of several physiopathological conditions. In chronic fatigue syndrome disturbances in autonomic activity, and in other homoeostatic mechanisms, such as the hormonal and immune systems, have been reported recently. In this review we followed the hypothesis that in chronic fatigue syndrome the paradoxical condition of disturbing somatic symptoms in the absence of organic evidence of disease might be addressed by focusing on attending functional correlates. In particular we addressed possible alterations in cardiovascular autonomic control, as can be assessed by spectral analysis of R-R interval and systolic arterial pressure variability. With this approach, in subjects complaining of unexplained fatigue, we obtained data suggesting a condition of prevailing sympathetic modulation of the sino-atrial node at rest, and reduced responsiveness to excitatory stimuli. Far from considering the issue resolved, we propose that in the context of the multiple physiological and psychological interactions involved in the perception and self-reporting of symptoms, attendant changes in physiological equivalents might furnish a convenient assessment independent from subjective components. Indices of sympathetic modulation could, accordingly, provide quantifiable signs of the interaction between subject's efforts and environmental demands, independently of self descriptions, which could provide convenient measurable outcomes, both for diagnosis and treatment titration in chronic fatigue syndrome. PMID- 9857115 TI - Can medicine prevent war? Imaginative thinking shows that it might. PMID- 9857116 TI - Reinventing doctors. Will move doctors from this winter of discontent to a position of leadership. PMID- 9857118 TI - Snapshots from the decade of the brain. Exciting, but no cause for triumphalism PMID- 9857117 TI - Socioeconomic differentials in the mortality of pets probably reflect the same differences in material circumstances as in their owners. PMID- 9857119 TI - "No longer Gage": an iron bar through the head. Early observations of personality change after injury to the prefrontal cortex. PMID- 9857120 TI - BMJ bans anniversaries. We promise our readers no more boring anniversaries PMID- 9857121 TI - Larks and owls and health, wealth, and wisdom. AB - OBJECTIVE: To test the validity of Benjamin Franklin's maxim "early to bed and early to rise makes a man healthy, wealthy, and wise." DESIGN: Cross sectional analysis of sleeping patterns in a nationally representative group of elderly people, and longitudinal investigation of mortality. SETTING: Eight areas in Britain (five in England, two in Scotland, and one in Wales). SUBJECTS: 1229 men and women aged 65 and over who in 1973-4 had taken part in a survey funded by the Department of Health and Social Security and for whom data on sleeping patterns, health, socioeconomic circumstances, and cognitive function had been recorded. MAIN OUTCOME MEASURES: Self reported income, access to a car, standard of accommodation, performance on a test of cognitive function, state of health and mortality during 23 years of follow up. RESULTS: 356 people (29%) were defined as larks (to bed before 11 pm and up before 8 am) and 318 (26%) were defined as owls (to bed at or after 11 pm and up at or after 8 am). There was no indication that larks were richer than those with other sleeping patterns. On the contrary, owls had the largest mean income and were more likely to have access to a car. There was also no evidence that larks were superior to those with other sleeping patterns with regard to their cognitive performance or their state of health. Both larks and owls had a slightly reduced risk of death compared with the rest of the study sample, but this was accounted for by the fact that they spent less time in bed at night. In the study sample as a whole, longer periods of time in bed were associated with increased mortality. After adjustment for age, sex, the presence of illness, and other risk factors, people who spent 12 or more hours in bed had a relative risk of death of 1.7 (1.2 to 2.5) compared with those who were in bed for 9 hours. The lowest risk occurred in people who spent 8 hours in bed (adjusted relative risk 0.8; 0.7 to 1.0). CONCLUSION: These findings do not support Franklin's claim. A "late to bed and late to rise" lifestyle does not seem to lead to socioeconomic, cognitive, or health disadvantage, but a longer time spent in bed may be associated with increased mortality. PMID- 9857122 TI - Does the fly matter? the CRACKPOT study in evidence based trout fishing. The Collaborative Randomised and Controlled Kennet Piscatorial Options Trail (CRACKPOT) Investigators. AB - OBJECTIVE: To investigate the importance of the type of dry fly (artificial floating fly) in catching trout (brown and rainbow) in an English chalkstream. SETTING: River Kennet, Berkshire. DESIGN: Five anglers on five separate occasions spent five hours using a randomly allocated fly from a sample of five types. PARTICIPANTS: Five anglers of considerable but varying experience, determination, and opinion. MAIN OUTCOME MEASURES: Number, weight, and species of trout caught. RESULTS: One fly (Black Gnat) performed significantly worse than the others. The fly most successful in catching brown trout was the Cinnamon Sedge. CONCLUSION: The possible prolongation of doctors' leisure time consequent on the use of unproductive trout flies has resource implications for the NHS. Urgent funding of a definitive, large multiriver trial is needed. PMID- 9857123 TI - Influence of superstition on the date of hospital discharge and medical cost in Japan: retrospective and descriptive study. AB - OBJECTIVES: To determine the influence of superstition about Taian (a lucky day) Butsumetsu (an unlucky day) on decision to leave hospital. To estimate the costs of the effect of this superstition. DESIGN: Retrospective and descriptive study. SETTING: University hospital in Kyoto, Japan. SUBJECTS: Patients who were discharged alive from Kyoto University Hospital from 1 April 1992 to 31 March 1995. MAIN OUTCOME MEASURES: Mean number, age, and hospital stay of patients discharged on each day of six day cycle. RESULTS: The mean number, age, and hospital stay of discharged patients were highest on Taian and lowest on Butsumetsu (25.8 v 19.3 patients/day, P=0.0001; 43.9 v 41.4 years, P=0.0001; and 43.1 v 33.3 days, P=0.0001 respectively). The effect of this difference on the hospital's costs was estimated to be 7.4 million yen (?31 000). CONCLUSION: The superstition influenced the decision to leave hospital, contributing to higher medical care costs in Japan. Although hospital stays need to be kept as short as possible to minimise costs, doctors should not ignore the possible psychological effects on patients' health caused by dismissing the superstition. PMID- 9857124 TI - Life is sweet: candy consumption and longevity. PMID- 9857125 TI - Grandmothers' role in preventing unnecessary accident and emergency attendances: cohort study. PMID- 9857126 TI - Xenotransmission of the socioeconomic gradient in health? A population based study. PMID- 9857128 TI - The physician's prayer PMID- 9857127 TI - Chewing gum for extradural haemorrhage. PMID- 9857129 TI - 1755 and all that: a historical primer of transmissible spongiform encephalopathy. PMID- 9857130 TI - Brainy mind. PMID- 9857131 TI - The consciousness of sight. PMID- 9857132 TI - Brain drugs of the future. PMID- 9857133 TI - Differences in mental abilities. PMID- 9857134 TI - The brain, circadian rhythms, and clock genes. PMID- 9857135 TI - Neurogenetic determinism and the new euphenics. PMID- 9857136 TI - An international work exchange: better than a midlife crisis. PMID- 9857137 TI - Only if she has a fishing rod! PMID- 9857138 TI - Apgar scores for dads. PMID- 9857139 TI - De futuro urbanorum. PMID- 9857140 TI - Alastair MacIntosh MacLachlainn (1892-1959): an unusual career. PMID- 9857141 TI - Get your book published. PMID- 9857143 TI - To the abattoir! PMID- 9857144 TI - Down with EBM! PMID- 9857142 TI - Lessons from Whitehall. PMID- 9857145 TI - The need for political correctness in scientific writing PMID- 9857146 TI - Double bandaging of sprained ankles. PMID- 9857147 TI - A seamless service. PMID- 9857148 TI - The primacy of primary health care. PMID- 9857149 TI - Clinical governance. PMID- 9857150 TI - The promise of the neurosciences. PMID- 9857151 TI - Medical omniscience. PMID- 9857152 TI - Doctors' autonomy. PMID- 9857154 TI - The prayer of the oppressed PMID- 9857153 TI - The accelerating pace of change. PMID- 9857155 TI - The ship PMID- 9857156 TI - What's in a name? PMID- 9857158 TI - Shall I compare thee to a PMID- 9857157 TI - A christmas carol PMID- 9857159 TI - Nan West's murals in the royal national orthopaedic hospital PMID- 9857161 TI - The new victorians: community care in the 1990s PMID- 9857160 TI - Coming to terms with death PMID- 9857162 TI - Will you present the prizes? PMID- 9857163 TI - My cancer and the magic bullet PMID- 9857164 TI - From the other side PMID- 9857166 TI - It's all right to get up late PMID- 9857165 TI - Boom ditty boom ditty boom ditty boom (goodness gracious me) PMID- 9857167 TI - A cinnamon sedge is better than a black gnat PMID- 9857168 TI - Superstition increases japanese health costs PMID- 9857169 TI - Sweets in moderation may be good for you PMID- 9857170 TI - Grandmothers prevent unnecessary hospital attendance PMID- 9857171 TI - Socioeconomic status affects the health of pets PMID- 9857172 TI - The ubiquitin-proteasome pathway: on protein death and cell life. PMID- 9857174 TI - Yeast mitochondrial F1F0-ATP synthase exists as a dimer: identification of three dimer-specific subunits. AB - Using the technique of blue native gel electrophoresis, the oligomeric state of the yeast mitochondrial F1F0-ATP synthase was analysed. Solubilization of mitochondrial membranes with low detergent to protein ratios led to the identification of the dimeric state of the ATP synthase. Analysis of the subunit composition of the dimer, in comparison with the monomer, revealed the presence of three additional small proteins. These dimer-specific subunits of the ATP synthase were identified as the recently described subunit e/Tim11 (Su e/Tim11), the putative subunit g homolog (Su g) and a new component termed subunit k (Su k). Although, as shown here, these three proteins are not required for the formation of enzymatically active ATP synthase, Su e/Tim11 and Su g are essential for the formation of the dimeric state. Su e/Tim11 appears to play a central role in this dimerization process. The dimer-specific subunits are associated with the membrane bound F0-sector. The F0-sector may thereby be involved in the dimerization of two monomeric F1F0-ATP synthase complexes. We speculate that the F1F0-ATP synthase of yeast, like the other complexes of oxidative phosphorylation, form supracomplexes to optimize transduction of energy and to enhance the stability of the complex in the membrane. PMID- 9857173 TI - A viral activator of gene expression functions via the ubiquitin-proteasome pathway. AB - The ability of herpes simplex virus type 1 (HSV-1) to attain a latent state in sensory neurones and reactivate periodically is crucial for its biological and clinical properties. The active transcription of the entire 152 kb viral genome during lytic replication contrasts with the latent state, which is characterized by the production of a single set of nuclear-retained transcripts. Reactivation of latent genomes to re-initiate the lytic cycle therefore involves a profound change in viral transcriptional activity, but the mechanisms by which this fundamentally important process occurs are yet to be well understood. In this report we show that the stimulation of the onset of viral lytic infection mediated by the viral immediate-early (IE) protein Vmw110 is strikingly inhibited by inactivation of the ubiquitin-proteasome pathway. Similarly, the Vmw110 dependent reactivation of quiescent viral genomes in cultured cells is also dependent on proteasome activity. These results constitute the first demonstration that the transcriptional activity of a viral genome can be regulated by protein stability control pathways. PMID- 9857175 TI - FRET microscopy demonstrates molecular association of non-specific lipid transfer protein (nsL-TP) with fatty acid oxidation enzymes in peroxisomes. AB - The fate of fluorescently labeled pre-nsL-TP (Cy3-pre-nsL-TP) microinjected into BALB/c 3T3 fibroblasts was investigated by confocal laser scanning microscopy. The protein exhibited a distinct punctate fluorescence pattern and colocalized to a high degree with the immunofluorescence pattern for the peroxisomal enzyme acyl CoA oxidase. Proteolytic removal of the C-terminal leucine of the putative peroxisomal targeting sequence (AKL) resulted in a diffuse cytosolic fluorescence. These results indicate that microinjected Cy3-pre-nsL-TP is targeted to peroxisomes. The association of nsL-TP with peroxisomal enzymes was investigated in cells by measuring fluorescence resonance energy transfer (FRET) between the microinjected Cy3-pre-nsL-TP and Cy5-labeled antibodies against the peroxisomal enzymes acyl-CoA oxidase, 3-ketoacyl-CoA thiolase, bifunctional enzyme, PMP70 and catalase. The technique of photobleaching digital imaging microscopy (pbDIM), used to quantitate the FRET efficiency on a pixel-by-pixel basis, revealed a specific association of nsL-TP with acyl-CoA oxidase, 3 ketoacyl-CoA thiolase and bifunctional enzyme in the peroxisomes. These observations were corroborated by subjecting a peroxisomal matrix protein fraction to affinity chromatography on Sepharose-immobilized pre-nsL-TP. Acyl-CoA oxidase was retained. These studies provide strong evidence for a role of nsL-TP in the regulation of peroxisomal fatty acid beta-oxidation, e.g. by facilitating the presentation of substrates and/or stabilization of the enzymes. PMID- 9857176 TI - Mutations in the pore regions of the yeast K+ channel YKC1 affect gating by extracellular K+. AB - The product of the Saccharomyces cerevisiae K+-channel gene YKC1 includes two pore-loop sequences that are thought to form the hydrophilic lining of the pore. Gating of the channel is promoted by membrane depolarization and is regulated by extracellular K+ concentration ([K+]o) both in the yeast and when expressed in Xenopus oocytes. Analysis of the wild-type current now shows that: (i) [K+]o suppresses a very slowly relaxing component, accelerating activation; (ii) [K+]o slows deactivation in a dose-dependent fashion; and (iii) Rb+, Cs+ and, to a lesser extent, Na+ substitute for K+ in its action on gating. We have identified single residues, L293 and A428, at equivalent positions within the two pore loops that affect the [K+]o sensitivity. Substitution of these residues gave channels with reduced sensitivity to [K+]o in macroscopic current kinetics and voltage dependence, but had only minor effects on selectivity among alkali cations in gating and on single-channel conductance. In some mutants, activation was slowed sufficiently to confer a sigmoidicity to current rise at low [K+]o. The results indicate that these residues are involved in [K+]o sensing. Their situation close to the permeation pathway points to an interaction between gating and permeation. PMID- 9857177 TI - Drosophila grim induces apoptosis in mammalian cells. AB - Genetic studies have shown that grim is a central genetic switch of programmed cell death in Drosophila; however, homologous genes have not been described in other species, nor has its mechanism of action been defined. We show here that grim expression induces apoptosis in mouse fibroblasts. Cell death induced by grim in mammalian cells involves membrane blebbing, cytoplasmic loss and nuclear DNA fragmentation. Grim-induced apoptosis is blocked by both natural and synthetic caspase inhibitors. We found that grim itself shows caspase-dependent proteolytic processing of its C-terminus in vitro. Grim-induced death is antagonized by bcl-2 in a dose-dependent manner, and neither Fas signalling nor p53 are required for grim pro-apoptotic activity. Grim protein localizes both in the cytosol and in the mitochondria of mouse fibroblasts, the latter location becoming predominant as apoptosis progresses. These results show that Drosophila grim induces death in mammalian cells by specifically acting on mitochondrial apoptotic pathways executed by endogenous caspases. These findings advance our knowledge of the mechanism by which grim induces apoptosis and show the conservation through evolution of this crucial programmed cell death pathway. PMID- 9857178 TI - A link between cell cycle and cell death: Bax and Bcl-2 modulate Cdk2 activation during thymocyte apoptosis. AB - Resting thymocytes undergoing apoptosis in response to specific stimuli degrade the cdk inhibitor p27(Kip1) and upregulate Cdk2 kinase activity. Inhibition of Cdk2 kinase activity efficiently blocks cell death via certain apoptosis pathways whereas overexpression of Cdk2 accelerates such cell death, suggesting its involvement in the signal transduction pathways activated by certain apoptotic stimuli. We found that Cdk2 activation during thymocyte apoptosis can be regulated by p53, Bax and Bcl-2. The highly elevated Cdk2 kinase activity in the apoptosing thymocytes is not associated with its canonical cyclins, cyclin E and cyclin A, and requires de novo synthesis of proteins for activation to take place. We therefore propose Cdk2 activation to be a crucial event in distinct pathways of apoptosis and the point at which the cell cycle and cell death pathways interact. PMID- 9857180 TI - Fission yeast Csk1 is a CAK-activating kinase (CAKAK). AB - Cell cycle progression is dependent on the sequential activity of cyclin dependent kinases (CDKs). For full activity, CDKs require an activating phosphorylation of a conserved residue (corresponding to Thr160 in human CDK2) carried out by the CDK-activating kinase (CAK). Two distinct CAK kinases have been described: in budding yeast Saccharomyces cerevisiae, the Cak1/Civ1 kinase is responsible for CAK activity. In several other species including human, Xenopus, Drosophila and fission yeast Schizosaccharomyces pombe, CAK has been identified as a complex homologous to CDK7-cyclin H (Mcs6-Mcs2 in fission yeast). Here we identify the fission yeast Csk1 kinase as an in vivo activating kinase of the Mcs6-Mcs2 CAK defining Csk1 as a CAK-activating kinase (CAKAK). PMID- 9857179 TI - Cdc6 protein causes premature entry into S phase in a mammalian cell-free system. AB - We exploit an improved mammalian cell-free DNA replication system to analyse quiescence and Cdc6 function. Quiescent 3T3 nuclei cannot initiate replication in S phase cytosol from HeLa or 3T3 cells. Following release from quiescence, nuclei become competent to initiate semiconservative DNA replication in S phase cytosol, but not in G0 phase cytosol. Immunoblots show that quiescent cells lack Cdc6 and that minichromosome maintenance (MCM) proteins are not associated with chromatin. Competence of G1 phase nuclei to replicate in vitro coincides with maximum Cdc6 accumulation and MCM protein binding to chromatin in vivo. Addition of recombinant Cdc6 to permeabilized, but not intact, G1 nuclei causes up to 82% of the nuclei to initiate and accelerates G1 progression, making nuclei competent to replicate prematurely. PMID- 9857181 TI - Analysis of Rad3 and Chk1 protein kinases defines different checkpoint responses. AB - Eukaryotic cells respond to DNA damage and S phase replication blocks by arresting cell-cycle progression through the DNA structure checkpoint pathways. In Schizosaccharomyces pombe, the Chk1 kinase is essential for mitotic arrest and is phosphorylated after DNA damage. During S phase, the Cds1 kinase is activated in response to DNA damage and DNA replication blocks. The response of both Chk1 and Cds1 requires the six 'checkpoint Rad' proteins (Rad1, Rad3, Rad9, Rad17, Rad26 and Hus1). We demonstrate that DNA damage-dependent phosphorylation of Chk1 is also cell-cycle specific, occurring primarily in late S phase and G2, but not during M/G1 or early S phase. We have also isolated and characterized a temperature-sensitive allele of rad3. Rad3 functions differently depending on which checkpoint pathway is activated. Following DNA damage, rad3 is required to initiate but not maintain the Chk1 response. When DNA replication is inhibited, rad3 is required for both initiation and maintenance of the Cds1 response. We have identified a strong genetic interaction between rad3 and cds1, and biochemical evidence shows a physical interaction is possible between Rad3 and Cds1, and between Rad3 and Chk1 in vitro. Together, our results highlight the cell-cycle specificity of the DNA structure-dependent checkpoint response and identify distinct roles for Rad3 in the different checkpoint responses. KEYWORDS: ATM/ATR/cell-cycle checkpoints/Chk1/Rad3 PMID- 9857182 TI - Low-affinity nerve-growth factor receptor (P75NTR) can serve as a receptor for rabies virus. AB - A random-primed cDNA expression library constructed from the mRNA of neuroblastoma cells (NG108) was used to clone a specific rabies virus (RV) receptor. A soluble form of the RV glycoprotein (Gs) was utilized as a ligand to detect positive cells. We identified the murine low-affinity nerve-growth factor receptor, p75NTR. BSR cells stably expressing p75NTR were able to bind Gs and G expressing lepidopteran cells. The ability of the RV glycoprotein to bind p75NTR was dependent on the presence of a lysine and arginine in positions 330 and 333 respectively of antigenic site III, which is known to control virus penetration into motor and sensory neurons of adult mice. P75NTR-expressing BSR cells were permissive for a non-adapted fox RV isolate (street virus) and nerve growth factor (NGF) decreased this infection. In infected cells, p75NTR associates with the RV glycoprotein and could be precipitated with anti-G monoclonal antibodies. Therefore, p75NTR is a receptor for street RV. PMID- 9857183 TI - A metalloprotease-disintegrin, MDC9/meltrin-gamma/ADAM9 and PKCdelta are involved in TPA-induced ectodomain shedding of membrane-anchored heparin-binding EGF-like growth factor. AB - The ectodomains of many proteins located at the cell surface are shed upon cell stimulation. One such protein is the heparin-binding EGF-like growth factor (HB EGF) that exists in a membrane-anchored form which is converted to a soluble form upon cell stimulation with TPA, an activator of protein kinase C (PKC). We show that PKCdelta binds in vivo and in vitro to the cytoplasmic domain of MDC9/meltrin-gamma/ADAM9, a member of the metalloprotease-disintegrin family. Furthermore, the presence of constitutively active PKCdelta or MDC9 results in the shedding of the ectodomain of proHB-EGF, whereas MDC9 mutants lacking the metalloprotease domain, as well as kinase-negative PKCdelta, suppress the TPA induced shedding of the ectodomain. These results suggest that MDC9 and PKCdelta are involved in the stimulus-coupled shedding of the proHB-EGF ectodomain. PMID- 9857184 TI - Mona, a novel hematopoietic-specific adaptor interacting with the macrophage colony-stimulating factor receptor, is implicated in monocyte/macrophage development. AB - The production, survival and function of monocytes and macrophages are regulated by the macrophage colony-stimulating factor (M-CSF or CSF-1) through its tyrosine kinase receptor Fms. Binding of M-CSF results in Fms autophosphorylation on specific tyrosines that act as docking sites for intracellular signaling molecules containing SH2 domains. Using a yeast two-hybrid screen, we cloned a novel adaptor protein which we called 'Mona' for monocytic adaptor. Mona contains one SH2 domain and two SH3 domains related to the Grb2 adaptor. Accordingly, Mona interacts with activated Fms on phosphorylated Tyr697, which is also the Grb2 binding site. Furthermore, Mona contains a unique proline-rich region located between the SH2 domain and the C-terminal SH3 domain, and is apparently devoid of any catalytic domain. Mona expression is restricted to two hematopoietic tissues: the spleen and the peripheral blood mononuclear cells, and is induced rapidly during monocytic differentiation of the myeloid NFS-60 cell line in response to M CSF. Strikingly, overexpression of Mona in bone marrow cells results in strong reduction of M-CSF-dependent macrophage production in vitro. Taken together, our results suggest an important role for Mona in the regulation of monocyte/macrophage development as controlled by M-CSF. PMID- 9857185 TI - A function-structure model for NGF-activated TRK. AB - Mechanisms regulating transit of receptor tyrosine kinases (RTKs) from inactive to active states are incompletely described, but require autophosphorylation of tyrosine(s) within a kinase domain 'activation loop'. Here, we employ functional biological assays with mutated TRK receptors to assess a 'switch' model for RTK activation. In this model: (i) ligand binding stimulates activation loop tyrosine phosphorylation; (ii) these phosphotyrosines form specific charge pairs with nearby basic residues; and (iii) the charge pairs stabilize a functionally active conformation in which the activation loop is restrained from blocking access to the kinase catalytic core. Our findings both support this model and identify residues that form specific charge pairs with each of the three TRK activation loop phosphotyrosines. PMID- 9857186 TI - Inactivation and dephosphorylation of protein kinase Balpha (PKBalpha) promoted by hyperosmotic stress. AB - To study the role of protein kinase B (PKB) in response to cellular stress, we examined PKBalpha activity following different stress treatments. Hyperosmotic but not chemical stress resulted in inactivation of PKBalpha and prevented activation by pervanadate and mitogens. Hyperosmotic shock did not affect the MAP kinase pathway, suggesting that this inhibitory effect was specific for PKB. Our data further indicate that downregulation occurs via dephosphorylation of Thr308 and Ser473, the major regulatory phosphorylation sites of PKBalpha. Indeed, calyculin A, which inhibits protein phosphatases 1 and 2A, effectively blocked hyperosmotic stress-mediated inactivation (dephosphorylation) of PKBalpha. High osmolarity did not affect phosphatidylinositol 3-kinase activity but led to a marked increase in PI(3,4,5)P3 and a decrease in PI(3,4)P2 formation after pervanadate stimulation, suggesting that hyperosmotic stress has an inhibitory effect on a phosphatidylinositol 5-phosphatase which converts PI(3,4,5)P3 into PI(3,4)P2. Immunofluorescence studies revealed that membrane translocation, a prerequisite for PKB activation, was not affected by hyperosmotic stress. Our results indicate that hyperosmotic stress can act at two levels: (i) inhibition of phosphorylation of Thr308 and Ser473 by upstream kinases and (ii) by promoting rapid dephosphorylation of these regulatory sites. PMID- 9857187 TI - Induction of the antigen receptor expression on B lymphocytes results in rapid competence for signaling of SLP-65 and Syk. AB - The binding of antigen to the B cell antigen receptor (BCR) results in the activation of protein tyrosine kinases (PTKs) such as Lyn and Syk, and the phosphorylation of several substrate proteins including HS1 and SLP-65. How these signaling elements are connected to the BCR is not well understood. Using an expression vector for a tamoxifen-regulated Cre recombinase, we have developed a method that allows the inducible expression of the BCR. Disruption of the VH leader reading frame of the immunoglobulin heavy chain by two loxP sites is overcome by Cre-mediated DNA recombination and results in the cell surface expression of the BCR starting 4 h after exposure of transfected B cells to tamoxifen. This method can, in principle, be employed for the inducible expression of any secreted or type I transmembrane protein. By monitoring the activation of signaling elements in pervanadate-stimulated B cells expressing different levels of the BCR, we show here that phosphorylation of SLP-65 and Syk, but not of Lyn, is strictly dependent on the expression of the BCR on the cell surface. These data suggest that the BCR reorganizes its signaling molecules as soon as it appears on the cell surface. PMID- 9857188 TI - Targeted disruption of SHIP leads to Steel factor-induced degranulation of mast cells. AB - To investigate the role of the src homology 2 (SH2)-containing inositol 5' phosphatase (SHIP) in growth factor-mediated signalling, we compared Steel factor (SF)-induced events in bone marrow-derived mast cells (BMMCs) from SHIP-/- and SHIP+/+ littermates. We found SF alone stimulated massive degranulation from SHIP /- but none from SHIP+/+ BMMCs. This SF-induced degranulation, which was not due to higher c-kit levels in SHIP-/- cells, correlated with higher intracellular calcium than that in SHIP+/+ cells and was dependent on the influx of extracellular calcium. Both this influx and subsequent degranulation were completely inhibited by PI-3-kinase inhibitors, indicating that SF-induced activation of PI-3-kinase was upstream of extracellular calcium entry. A comparison of phosphatidylinositol-3,4,5-trisphosphate (PIP3) levels following SF stimulation of SHIP+/+ and SHIP-/- BMMCs suggested that SHIP restricted this entry by hydrolyzing PIP3. Although PI-3-kinase inhibitors blocked the release of intracellular calcium, implicating PIP3, and PLCgamma-2 was slightly more tyrosine phosphorylated in SHIP-/- cells, the increase in inositol-1,4,5 trisphosphate (IP3) and intracellular calcium levels were identical in SHIP-/- and SHIP+/+ BMMCs. These results suggest that SHIP prevents SF from triggering degranulation of normal BMMCs, and does so by hydrolyzing PIP3, which in turn limits extracellular calcium entry at a step after the release of intracellular calcium. PMID- 9857189 TI - A cdc15-like adaptor protein (CD2BP1) interacts with the CD2 cytoplasmic domain and regulates CD2-triggered adhesion. AB - A human CD2 cytoplasmic tail-binding protein, termed CD2BP1, was identified by an interaction trap cloning method. Expression of CD2BP1 is restricted to hematopoietic tissue, being prominent in T and natural killer (NK) cells, with long (CD2BP1L) and short (CD2BP1S) variants arising by alternative RNA splicing. Both CD2BP1 molecules are homologous to Schizosaccharomyces pombe cdc15, and include a helical domain, variable length intervening PEST sequence and C terminal SH3 domain. Although the CD2BP1 SH3 domain binds directly to the CD2 sequence, KGPPLPRPRV (amino acids 300-309), its association is augmented markedly by the CD2BP1 N-terminal segment. Upon ligand-induced clustering of surface CD2 molecules, CD2BP1 redistributes from a cytosolic to a surface membrane compartment, co-localizing with CD2. In turn, CD2-stimulated adhesion is downregulated by CD2BP1, apparently through coupling of the protein tyrosine phosphatase (PTP)-PEST to CD2. These findings offer the first molecular view into the control processes for T cell adhesion. PMID- 9857190 TI - PTP-SL and STEP protein tyrosine phosphatases regulate the activation of the extracellular signal-regulated kinases ERK1 and ERK2 by association through a kinase interaction motif. AB - Protein kinases and phosphatases regulate the activity of extracellular signal regulated kinases 1 and 2 (ERK1/2) by controlling the phosphorylation of specific residues. We report the physical and functional association of ERK1/2 with the PTP-SL and STEP protein tyrosine phosphatases (PTPs). Upon binding, the N terminal domains of PTP-SL and STEP were phosphorylated by ERK1/2, whereas these PTPs dephosphorylated the regulatory phosphotyrosine residues of ERK1/2 and inactivated them. A sequence of 16 amino acids in PTP-SL was identified as being critical for ERK1/2 binding and termed kinase interaction motif (KIM) (residues 224-239); it was shown to be required for phosphorylation of PTP-SL by ERK1/2 at Thr253. Co-expression of ERK2 with catalytically active PTP-SL in COS-7 cells impaired the EGF-induced activation of ERK2, whereas a PTP-SL mutant, lacking PTP activity, increased the ERK2 response to EGF. This effect was dependent on the presence of the KIM on PTP-SL. Furthermore, ERK1/2 activity was downregulated in 3T3 cells stably expressing PTP-SL. Our findings demonstrate the existence of a conserved ERK1/2 interaction motif within the cytosolic non-catalytic domains of PTP-SL and STEP, which is required for the regulation of ERK1/2 activity and for phosphorylation of the PTPs by these kinases. Our findings suggest that PTP-SL and STEP act as physiological regulators of the ERK1/2 signaling pathway. PMID- 9857191 TI - Notchless encodes a novel WD40-repeat-containing protein that modulates Notch signaling activity. AB - Signaling by Notch family receptors is involved in many cell-fate decisions during development. Several modifiers of Notch activity have been identified, suggesting that regulation of Notch signaling is complex. In a genetic screen for modifiers of Notch activity, we identified a gene encoding a novel WD40-repeat protein. The gene is called Notchless, because loss-of-function mutant alleles dominantly suppress the wing notching caused by certain Notch alleles. Reducing Notchless activity increases Notch activity. Overexpression of Notchless in Xenopus or Drosophila appears to have a dominant-negative effect in that it also increases Notch activity. Biochemical studies show that Notchless binds to the cytoplasmic domain of Notch, suggesting that it serves as a direct regulator of Notch signaling activity. PMID- 9857192 TI - Regionalized metabolic activity establishes boundaries of retinoic acid signalling. AB - The competence of a cell to respond to the signalling molecule retinoic acid (RA) is thought to depend largely on its repertoire of cognate zinc finger nuclear receptors. XCYP26 is an RA hydroxylase that is expressed differentially during early Xenopus development. In Xenopus embryos, XCYP26 can rescue developmental defects induced by application of exogenous RA, suggesting that the enzymatic modifications introduced inhibit RA signalling activities in vivo. Alterations in the expression pattern of a number of different molecular markers for neural development induced upon ectopic expression of XCYP26 reflect a primary function of RA signalling in hindbrain development. Progressive inactivation of RA signalling results in a stepwise anteriorization of the molecular identity of individual rhombomeres. The expression pattern of XCYP26 during gastrulation appears to define areas within the prospective neural plate that develop in response to different concentrations of RA. Taken together, these observations appear to reflect an important regulatory function of XCYP26 for RA signalling; XCYP26-mediated modification of RA modulates its signalling activity and helps to establish boundaries of differentially responsive and non-responsive territories. PMID- 9857193 TI - Mitotic silencing of human rRNA synthesis: inactivation of the promoter selectivity factor SL1 by cdc2/cyclin B-mediated phosphorylation. AB - We have used a reconstituted cell-free transcription system to investigate the molecular basis of mitotic repression of RNA polymerase I (pol I) transcription. We demonstrate that SL1, the TBP-containing promoter-binding factor, is inactivated by cdc2/cyclin B-directed phosphorylation, and reactivated by dephosphorylation. Transcriptional inactivation in vitro is accompanied by phosphorylation of two subunits, e.g. TBP and hTAFI110. To distinguish whether transcriptional repression is due to phosphorylation of TBP, hTAFI110 or both, SL1 was purified from two HeLa cell lines that express either full-length or the core domain of TBP only. Both TBP-TAFI complexes exhibit similar activity and both are repressed at mitosis, indicating that the variable N-terminal domain which contains multiple target sites for cdc2/cyclin B phosphorylation is dispensable for mitotic repression. Protein-protein interaction studies reveal that mitotic phosphorylation impairs the interaction of SL1 with UBF. The results suggest that phosphorylation of SL1 is used as a molecular switch to prevent pre initiation complex formation and to shut down rDNA transcription at mitosis. PMID- 9857194 TI - Leukemic transformation by the v-ErbA oncoprotein entails constitutive binding to and repression of an erythroid enhancer in vivo. AB - v-ErbA, a mutated thyroid hormone receptor alpha (TRalpha), is thought to contribute to avian erythroblastosis virus (AEV)-induced leukemic transformation by constitutively repressing transcription of target genes. However, the binding of v-ErbA or any unliganded nuclear receptor to a chromatin-embedded response element as well as the role of the N-CoR-SMRT-HDAC co-repressor complex in mediating repression remain hypothetical. Here we identify a v-ErbA-response element, VRE, in an intronic DNase I hypersensitive site (HS2) of the chicken erythroid carbonic anhydrase II (CAII) gene. In vivo footprinting shows that v ErbA is constitutively bound to this HS2-VRE in transformed, undifferentiated erythroblasts along with other transcription factors like GATA-1. Transfection assays show that the repressed HS2 region can be turned into a potent enhancer in v-ErbA-expressing cells by mutation of the VRE. Differentiation of transformed cells alleviates v-ErbA binding concomitant with activation of CAII transcription. Co-expression of a gag-TRalpha fusion protein in AEV-transformed cells and addition of ligand derepresses CAII transcription. Treatment of transformed cells with the histone deacetylase inhibitor, trichostatin A, derepresses the endogenous, chromatin-embedded CAII gene, while a transfected HS2 enhancer construct remains repressed. Taken together, our data suggest that v ErbA prevents CAII activation by 'neutralizing' in cis the activity of erythroid transcription factors. PMID- 9857195 TI - Evidence that P-TEFb alleviates the negative effect of DSIF on RNA polymerase II dependent transcription in vitro. AB - Recently, a positive and a negative elongation factor, implicated in 5,6-dichloro 1-beta-D-ribofuranosylbenzimidazole (DRB) inhibition of transcription elongation, has been identified. P-TEFb is a positive transcription elongation factor and the DRB-sensitive kinase that phosphorylates the C-terminal domain (CTD) of the largest subunit of RNA polymerase II (Pol II). PITALRE, a member of the Cdc2 family of protein kinases, is the catalytic subunit of P-TEFb. DSIF is a human homolog of the yeast Spt4-Spt5 complex and renders elongation of transcription sensitive to DRB. DRB sensitivity-inducing factor (DSIF) binds to RNA Pol II and may directly regulate elongation. Here we show a functional interaction between P TEFb and DSIF. The reduction of P-TEFb activity induced by either DRB, antibody against PITALRE, or immunodepletion resulted in a negative effect of DSIF on transcription. DSIF acts at an early phase of elongation, and the prior action of P-TEFb makes transcription resistant to DSIF. The state of phosphorylation of CTD determines the DSIF-RNA Pol II interaction, and may provide a direct link between P-TEFb and DSIF. Taken together, this study reveals a molecular basis for DRB action and suggests that P-TEFb stimulates elongation by alleviating the negative action of DSIF. PMID- 9857196 TI - The structure of plasmid-encoded transcriptional repressor CopG unliganded and bound to its operator. AB - The structure of the 45 amino acid transcriptional repressor, CopG, has been solved unliganded and bound to its target operator DNA. The protein, encoded by the promiscuous streptococcal plasmid pMV158, is involved in the control of plasmid copy number. The structure of this protein repressor, which is the shortest reported to date and the first isolated from a plasmid, has a homodimeric ribbon-helix-helix arrangement. It is the prototype for a family of homologous plasmid repressors. CopG cooperatively associates, completely protecting several turns on one face of the double helix in both directions from a 13-bp pseudosymmetric primary DNA recognition element. In the complex structure, one protein tetramer binds at one face of a 19-bp oligonucleotide, containing the pseudosymmetric element, with two beta-ribbons inserted into the major groove. The DNA is bent 60 degrees by compression of both major and minor grooves. The protein dimer displays topological similarity to Arc and MetJ repressors. Nevertheless, the functional tetramer has a unique structure with the two vicinal recognition ribbon elements at a short distance, thus inducing strong DNA bend. Further structural resemblance is found with helix-turn-helix regions of unrelated DNA-binding proteins. In contrast to these, however, the bihelical region of CopG has a role in oligomerization instead of DNA recognition. This observation unveils an evolutionary link between ribbon-helix-helix and helix turn-helix proteins. PMID- 9857197 TI - Crm1p mediates regulated nuclear export of a yeast AP-1-like transcription factor. AB - The yeast AP-1-like transcription factor, Yap1p, activates genes required for the response to oxidative stress. Yap1p is normally cytoplasmic and inactive, but will activate by nuclear translocation if cells are placed in an oxidative environment. Here we show that Yap1p is a target of the beta-karyopherin-like nuclear exporter, Crm1p. Yap1p is constitutively nuclear in a crm1 mutant, and Crm1p binds to a nuclear export sequence (NES)-like sequence in Yap1p in the presence of RanGTP. Recognition of Yap1p by Crm1p is inhibited by oxidation, and this inhibition requires at least one of the three cysteine residues flanking the NES. These results suggest that Yap1p localization is largely regulated at the level of nuclear export, and that the oxidation state affects the accessibility of the Yap1p NES to Crm1p directly. We also show that a mutation in RanGAP (rna1 1) is synthetically lethal with crm1 mutants. Yap1p export is inhibited in both rna1-1 and prp20 (RanGNRF) mutant strains, but Yap1p rapidly accumulates at the nuclear periphery after shifting rna1-1, but not other mutant cells to the non permissive temperature. Thus, disassembly of export complexes in response to RanGTP hydrolysis may be required for release of substrate from a terminal binding site at the nuclear pore complex (NPC). PMID- 9857198 TI - The role of exportin-t in selective nuclear export of mature tRNAs. AB - Exportin-t (Xpo-t) is a vertebrate nuclear export receptor for tRNAs that binds tRNA cooperatively with GTP-loaded Ran. Xpo-t antibodies are shown to efficiently block tRNA export from Xenopus oocyte nuclei suggesting that it is responsible for at least the majority of tRNA export in these cells. We examine the mechanism by which Xpo-t-RanGTP specifically exports mature tRNAs rather than other forms of nuclear RNA, including tRNA precursors. Chemical and enzymatic footprinting together with phosphate modification interference reveals an extensive interaction between the backbone of the TPsiC and acceptor arms of tRNAPhe and Xpo-t-RanGTP. Analysis of mutant or precursor tRNA forms demonstrates that, aside from these recognition elements, accurate 5' and 3' end-processing of tRNA affects Xpo-t-RanGTP interaction and nuclear export, while aminoacylation is not essential. Intron-containing, end-processed, pre-tRNAs can be bound by Xpo-t RanGTP and are rapidly exported from the nucleus if Xpo-t is present in excess. These results suggest that at least two mechanisms are involved in discrimination of pre-tRNAs and mature tRNAs prior to nuclear export. PMID- 9857199 TI - A role for the yeast La protein in U6 snRNP assembly: evidence that the La protein is a molecular chaperone for RNA polymerase III transcripts. AB - The first protein that binds to all newly synthesized RNA polymerase III transcripts is a highly conserved phosphoprotein known as the La autoantigen. Although binding by the yeast La protein Lhp1p to pre-tRNAs is required for the normal pathway of tRNA maturation, the role of the La protein in the biogenesis of other polymerase III transcripts has been unclear. We identified a mutation in a novel component of the U6 snRNP that causes yeast cells to require Lhp1p for growth. This protein, Lsm8p, is a member of a family of proteins, known as Sm like proteins, that shares two conserved motifs with the core Sm proteins of the U1, U2, U4 and U5 snRNPs. The lsm8-1 cells have drastically reduced levels of the mature U6 snRNP, consistent with a defect in U6 snRNP assembly. In these cells, Lhp1p stabilizes newly synthesized U6 RNA, thus facilitating assembly of the RNA into the U6 snRNP. These results provide evidence that Lhp1p is a molecular chaperone for polymerase III-transcribed RNAs and implicate Lsm8p as a key component in the very early steps of U6 snRNP assembly. PMID- 9857200 TI - Control of cleavage site selection during mRNA 3' end formation by a yeast hnRNP. AB - Endonucleolytic cleavage of pre-mRNAs is the first step during eukaryotic mRNA 3' end formation. It has been proposed that cleavage factors CF IA, CF IB and CF II are required for pre-mRNA 3' end cleavage in yeast. CF IB is composed of a single polypeptide, Nab4p/Hrp1p, which is related to the A/B group of metazoan heterogeneous nuclear ribonucleoproteins (hnRNPs) that function as antagonistic regulators of 5' splice site selection. Here, we provide evidence that Nab4p/Hrp1p is not required for pre-mRNA 3' end endonucleolytic cleavage. We show that CF IA and CF II devoid of Nab4p/Hrp1p are sufficient to cleave a variety of RNA substrates but that cleavage occurs at multiple sites. Addition of Nab4p/Hrp1p prevents these alternative cleavages in a concentration-dependent manner, suggesting an essential and conserved role for some hnRNPs in pre-mRNA cleavage site selection. PMID- 9857201 TI - Structure of the foot-and-mouth disease virus leader protease: a papain-like fold adapted for self-processing and eIF4G recognition. AB - The leader protease of foot-and-mouth disease virus, as well as cleaving itself from the nascent viral polyprotein, disables host cell protein synthesis by specific proteolysis of a cellular protein: the eukaryotic initiation factor 4G (eIF4G). The crystal structure of the leader protease presented here comprises a globular catalytic domain reminiscent of that of cysteine proteases of the papain superfamily, and a flexible C-terminal extension found intruding into the substrate-binding site of an adjacent molecule. Nevertheless, the relative disposition of this extension and the globular domain to each other supports intramolecular self-processing. The different sequences of the two substrates cleaved during viral replication, the viral polyprotein (at LysLeuLys/GlyAlaGly) and eIF4G (at AsnLeuGly/ArgThrThr), appear to be recognized by distinct features in a narrow, negatively charged groove traversing the active centre. The structure illustrates how the prototype papain fold has been adapted to the requirements of an RNA virus. Thus, the protein scaffold has been reduced to a minimum core domain, with the active site being modified to increase specificity. Furthermore, surface features have been developed which enable C-terminal self processing from the viral polyprotein. PMID- 9857202 TI - A newly identified N-terminal amino acid sequence of human eIF4G binds poly(A) binding protein and functions in poly(A)-dependent translation. AB - Most eukaryotic mRNAs possess a 5' cap and a 3' poly(A) tail, both of which are required for efficient translation. In yeast and plants, binding of eIF4G to poly(A)-binding protein (PABP) was implicated in poly(A)-dependent translation. In mammals, however, there has been no evidence that eIF4G binds PABP. Using 5' rapid amplification of cDNA, we have extended the known human eIF4GI open reading frame from the N-terminus by 156 amino acids. Co-immunoprecipitation experiments showed that the extended eIF4GI binds PABP, while the N-terminally truncated original eIF4GI cannot. Deletion analysis identified a 29 amino acid sequence in the new N-terminal region as the PABP-binding site. The 29 amino acid stretch is almost identical in eIF4GI and eIF4GII, and the full-length eIF4GII also binds PABP. As previously shown for yeast, human eIF4G binds to a fragment composed of RRM1 and RRM2 of PABP. In an in vitro translation system, an N-terminal fragment which includes the PABP-binding site inhibits poly(A)-dependent translation, but has no effect on translation of a deadenylated mRNA. These results indicate that, in addition to a recently identified mammalian PABP-binding protein, PAIP-1, eIF4G binds PABP and probably functions in poly(A)-dependent translation in mammalian cells. PMID- 9857203 TI - Complete kinetic mechanism of elongation factor Tu-dependent binding of aminoacyl tRNA to the A site of the E. coli ribosome. AB - The kinetic mechanism of elongation factor Tu (EF-Tu)-dependent binding of Phe tRNAPhe to the A site of poly(U)-programmed Escherichia coli ribosomes has been established by pre-steady-state kinetic experiments. Six steps were distinguished kinetically, and their elemental rate constants were determined either by global fitting, or directly by dissociation experiments. Initial binding to the ribosome of the ternary complex EF-Tu.GTP.Phe-tRNAPhe is rapid (k1 = 110 and 60/micromM/s at 10 and 5 mM Mg2+, 20 degreesC) and readily reversible (k-1 = 25 and 30/s). Subsequent codon recognition (k2 = 100 and 80/s) stabilizes the complex in an Mg2+-dependent manner (k-2 = 0.2 and 2/s). It induces the GTPase conformation of EF-Tu (k3 = 500 and 55/s), instantaneously followed by GTP hydrolysis. Subsequent steps are independent of Mg2+. The EF-Tu conformation switches from the GTP- to the GDP-bound form (k4 = 60/s), and Phe-tRNAPhe is released from EF-Tu.GDP. The accommodation of Phe-tRNAPhe in the A site (k5 = 8/s) takes place independently of EF-Tu and is followed instantaneously by peptide bond formation. The slowest step is dissociation of EF-Tu.GDP from the ribosome (k6 = 4/s). A characteristic feature of the mechanism is the existence of two conformational rearrangements which limit the rates of the subsequent chemical steps of A-site binding. PMID- 9857204 TI - Structure of the 3'-hairpin of the TYMV pseudoknot: preformation in RNA folding. AB - The solution structure of an RNA-hairpin present in the pseudoknot, which is found at the 3'-terminus of turnip yellow mosaic virus genomic RNA, has been solved by nuclear magnetic resonance spectroscopy. The loop, which contains the sequence 5'-GGGUCA-3', was found to be highly structured and, contrary to expectations, does not attain its stability through GA or GC base pair formation but by triple interactions between the tilted adenosine and the minor groove sides of the first two guanosines. Interestingly, a very similar conformation was found for the cognate pseudoknot, implying that the 3'-hairpin is preformed for folding into a pseudoknotted structure. These findings suggest a mechanism of 'predetermined-fit' as a principle in RNA folding. PMID- 9857205 TI - Molecular basis of double-stranded RNA-protein interactions: structure of a dsRNA binding domain complexed with dsRNA. AB - Protein interactions with double-stranded RNA (dsRNA) are critical for many cell processes; however, in contrast to protein-dsDNA interactions, surprisingly little is known about the molecular basis of protein-dsRNA interactions. A large and diverse class of proteins that bind dsRNA do so by utilizing an approximately 70 amino acid motif referred to as the dsRNA-binding domain (dsRBD). We have determined a 1.9 A resolution crystal structure of the second dsRBD of Xenopus laevis RNA-binding protein A complexed with dsRNA. The structure shows that the protein spans 16 bp of dsRNA, interacting with two successive minor grooves and across the intervening major groove on one face of a primarily A-form RNA helix. The nature of these interactions explains dsRBD specificity for dsRNA (over ssRNA or dsDNA) and the apparent lack of sequence specificity. Interestingly, the dsRBD fold resembles a portion of the conserved core structure of a family of polynucleotidyl transferases that includes RuvC, MuA transposase, retroviral integrase and RNase H. Structural comparisons of the dsRBD-dsRNA complex and models proposed for polynucleotidyl transferase-nucleic acid complexes suggest that similarities in nucleic acid binding also exist between these families of proteins. PMID- 9857207 TI - Chondrocyte differentiation in a rat mesenchymal cell line. AB - We used a combination of morphologic and histochemical methods to demonstrate that rat calvaria-derived mesenchymal cells, RCJ 3.1C5. 18, in culture progress through the differentiation pathway exhibited by chondrocytes in the endochondral growth plate. The cells were grown either as monolayer or suspension cultures. Subconfluent monolayer cultures did not express markers typical of chondrocyte phenotypes. However, after reaching confluency the cells formed nodules of chondrocytic cells separated by cartilage-appearing matrix and encapsulated by fibroblast-like cells. Suspension culture produced cell aggregates with similar characteristics. Matrix in both the nodules and aggregates stained for collagen Types II and XI and aggrecan, and some cells displayed a distinctive pericellular matrix that stained for Type X collagen. Mineralization was evident in older cultures. By electron microscopy, most cells in the aggregates appeared as typical chondrocytes. However, some larger cells were surrounded by a "mat" of matrix comprised of hexagonal arrays of dense nodules interconnected by a filamentous network. Immunogold localization confirmed the presence of collagen Type X in this matrix. Analysis of markers of chondrocyte differentiation and terminal differentiation over time showed that these markers were acquired sequentially over 2 weeks of culture. This model system will be useful to study the regulation of various steps in the chondrocyte differentiation pathway. PMID- 9857206 TI - Crystal structures of open and closed forms of binary and ternary complexes of the large fragment of Thermus aquaticus DNA polymerase I: structural basis for nucleotide incorporation. AB - The crystal structures of two ternary complexes of the large fragment of Thermus aquaticus DNA polymerase I (Klentaq1) with a primer/template DNA and dideoxycytidine triphosphate, and that of a binary complex of the same enzyme with a primer/template DNA, were determined to a resolution of 2.3, 2.3 and 2.5 A, respectively. One ternary complex structure differs markedly from the two other structures by a large reorientation of the tip of the fingers domain. This structure, designated 'closed', represents the ternary polymerase complex caught in the act of incorporating a nucleotide. In the two other structures, the tip of the fingers domain is rotated outward by 46 degrees ('open') in an orientation similar to that of the apo form of Klentaq1. These structures provide the first direct evidence in DNA polymerase I enzymes of a large conformational change responsible for assembling an active ternary complex. PMID- 9857208 TI - Immunohistochemical localization of Ca2+/Calmodulin-dependent protein kinase IV in outer hair cells. AB - A smooth membrane system consisting of subsurface cisternae (SSC) underlies the lateral plasmalemma of auditory outer hair cells (OHCs). The SSC contain Ca ATPase and are regarded as an intracellular Ca2+ reservoir like the sarcoplasmic reticulum of myocytes. Recently, it has been demonstrated that Ca-ATPase activity in sarcoplasmic reticulum is regulated by Ca2+/calmodulin-dependent protein kinases (CaM kinases). Here we investigated the presence of CaM kinases in OHCs and their possible association with the SSC. Inner ears collected from adult gerbils and from neonates at 2-day intervals between 0 and 20 days after birth were immunostained with antibodies specific for different CaM kinases. A polyclonal antiserum against CaM kinase IV yielded a strong immunostaining reaction along the lateral wall of OHCs. The staining appeared after the tenth postnatal day and continued into adulthood. No other site in the inner ear, including cochlear inner hair cells and vestibular hair cells, was reactive. The kinase's apparent association with the SSC strongly supports its involvement in intracellular Ca2+ homeostasis and suggests a role in regulating the OHCs' slow motile responses. PMID- 9857209 TI - Microwave staining of enteric neurons using cuprolinic blue (Quinolinic phthalocyanin) combined with enzyme histochemistry and peroxidase immunohistochemistry. AB - Methods that visualize subsets as well as the entire enteric neuron population are not readily available or have proved to be unreliable. Therefore, we attempted to combine NADPH-d histochemistry, AChE histochemistry, and CGRP immunohistochemistry, techniques that mark subsets of enteric neurons, with a technique that appeared to visualize the entire enteric neuron population, the cuprolinic blue staining method. To guarantee representative staining results, the individual staining methods were modified by using microwaves. In addition, this preserved the characteristics of each of the individual techniques. The distribution of NADPH-d, AChE, and CGRP corresponded well with previous morphological and physiological reports. Consequently, the different combinations gave rise to rapid, useful, and ready-to-use double labeling techniques. Their main advantage is that they simultaneously visualize the total population as well as subsets of enteric neurons. PMID- 9857210 TI - Fibroblast growth factor promotes recruitment of skeletal muscle satellite cells in young and old rats. AB - Although the role of satellite cells in muscle growth and repair is well recognized, understanding of the molecular events that accompany their activation and proliferation is limited. In this study, we used the single myofiber culture model for comparing the proliferative dynamics of satellite cells from growing (3 week-old), young adult (8- to 10-week-old), and old (9- to 11-month-old) rats. In these fiber cultures, the satellite cells are maintained in their in situ position underneath the fiber basement membrane. We first demonstrate that the cytoplasm of fiber-associated satellite cells can be monitored with an antibody against the extracellular signal regulated kinases 1 and 2 (ERK1 and ERK2), which belong to the mitogen-activated protein kinase (MAPK) superfamily. With this immunocytological marker, we show that the satellite cells from all three age groups first proliferate and express PCNA and MyoD, and subsequently, about 24 hr later, exit the PCNA+/MyoD+ state and become positive for myogenin. For all three age groups, fibroblast growth factor 2 (FGF2) enhances by about twofold the number of satellite cells that are capable of proliferation, as determined by monitoring the number of cells that transit from the MAPK+ phenotype to the PCNA+/MAPK+ or MyoD+/MAPK+ phenotype. Furthermore, contrary to the commonly accepted convention, we show that in the fiber cultures FGF2 does not suppress the subsequent transition of the proliferating cells into the myogenin+ compartment. Although myogenesis of satellite cells from growing, young adult, and old rats follows a similar program, two distinctive features were identified for satellite cells in fiber cultures from the old rats. First, a large number of MAPK+ cells do not appear to enter the MyoD-myogenin expression program. Second, the maximal number of proliferating satellite cells is attained a day later than in cultures from the young adults. This apparent "lag" in proliferation was not affected by hepatocyte growth factor (HGF), which has been implicated in accelerating the first round of satellite cell proliferation. HGF and FGF2 were equally efficient in promoting proliferation of satellite cells in fibers from old rats. Collectively, the investigation suggests that FGF plays a critical role in the recruitment of satellite cells into proliferation. PMID- 9857211 TI - Carbonic anhydrase in the gills of seawater- and freshwater-acclimated flounders Platichthys flesus: purification, characterization, and immunohistochemical localization. AB - Flounders Platichthys flesus were investigated with respect to isolation, purification, and cellular localization of carbonic anhydrase (CA) in the respiratory system. CA was purified from gills and erythrocytes and was shown to exclusively represent a soluble enzyme with an apparent molecular weight of 30 kD. Inhibition constants (KI) towards acetazolamide (ACTZ) were 8.4.10(-9) M for erythrocyte CA and 7.6.10(-9) M for gill CA, indicating a high sensitivity to sulfonamides, as exhibited by human CA II. Specific CA activity did not differ significantly in seawater- and freshwater-acclimated fish. Antibodies were raised against purified gill and erythrocyte CA. Both antisera crossreacted and were used to localize CA in the gills of seawater and freshwater flounders at the light microscopic level. Independent of the salinity, a positive reaction of variable intensity was found in the following cell types: pavement cells (PVCs), forming the gill epithelial surface layer; mucous cells (MCs); pillar cells (PCs), bordering the vascular channels of the secondary lamellae; and chloride cells (CCs), mitochondria-rich cells located in the primary epithelium, the interlamellar regions, and at the bases of the secondary lamellae.(J Histochem Cytochem 47:43-50, 1999) PMID- 9857212 TI - Differential expression of RNA and protein of the three pore-forming subunits of the amiloride-sensitive epithelial sodium channel in taste buds of the rat. AB - Salt taste signals from the rat anterior tongue are probably transduced via epithelial sodium channels (ENaCs) residing in the apical cellular pole of taste cells. The signals are blocked by mucosal amiloride in low microM concentrations. In contrast, the rat vallate papilla does not contribute to amiloride-blockable salt taste. Two approaches were used to probe for the three subunits of ENaC in the anterior and posterior tongue of the rats in sodium balance. (a) Immunohistochemistry with antibodies against ENaC subunits and against amiloride binding sites. In the anterior tongue, reactivity for alpha-, beta-, and gamma subunits was present in taste buds and lingual epithelium. In the posterior tongue vallate papilla, reactivity for alpha-subunit and for amiloride binding sites was easily demonstrable, whereas that for beta-subunit and especially for gamma-subunit was weaker than in the anterior tongue. (b) RT-PCR techniques were used to probe for the presence of ENaC subunit mRNA. In isolated taste buds of the anterior tongue, mRNA of all three subunits was found, whereas in isolated taste buds of the vallate papilla only mRNA of the alpha-subunit was easily detectable. That of beta- and gamma-subunits was much less abundant. RNA of all three subunits was abundant only in taste buds of the anterior tongue. Therefore, subsets of elongated taste cells do express ENaC, but regional differences exist in the transcription and expression of subunits. The regional differences suggest that amiloride-sensitive salt taste, which requires all three subunits, is present in the anterior but not the posterior tongue of rats, as functional studies indicate. PMID- 9857213 TI - Cytoskeletal and adhesive structural polarizations accompany IL-13-induced human macrophage fusion. AB - During the inflammatory response to an implanted biomaterial, monocytes undergo a striking phenotypic progression of differentiation into macrophages, which may subsequently fuse to form foreign body giant cells (FBGCs). Taking advantage of an in vitro system of cytokine-induced FBGC formation together with the optical slicing capabilities of a confocal microscope, we investigated the cytoskeletal reorganization and adhesive structure development during this dramatic morphological progression. Human monocytes demonstrated diffuse cytoplasmic staining of adhesive structural proteins. Punctate filamentous (F)-actin structures appeared along the ventral cell membrane of macrophages and were identified as the core of podosome adhesive structures by the distinctive ring staining of vinculin, talin, and paxillin around the F-actin. Cytokine-induced FBGCs were characterized by a restriction of podosomes to the extreme periphery of the ventral cell surface. Although macrophages and FBGC contained equivalent amounts of F-actin, significantly more F-actin was located within 1 micron of the ventral plasma membrane in FBGCs compared to macrophages. Taken together, these results provide new information on the dynamic cytoskeletal reorganization and adhesive structure development that occur during phenotypic progression from human monocytes to macrophages to FBGC. Furthermore, they suggest the acquisition of functional specializations on FBGC formation, which may enhance our understanding of chronic inflammatory processes. PMID- 9857214 TI - Two domains of rat galectin-4 bind to distinct structures of the intercellular borders of colorectal epithelia. AB - Galectin-4 (G4) is a member of a family of soluble galactoside-binding lectins found in various mammalian tissues. To determine the function of this protein in colorectal tissue, we separately produced the N- and C-terminal carbohydrate binding domains (CBD) of rat G4 as a recombinant glutathione S-transferase (GST) fusion protein (G4-N and G4-C) and examined the tissue binding site(s) of each CBD by light and electron microscopy (LM and EM). At the LM level, both fusion proteins stained the intercellular borders of the surface-lining epithelial cells of colorectal mucosa. At the EM level, two proteins recognized spatially close but distinct subcellular structures. G4-N stained electron-lucent flocculent substances freely located in the intercellular spaces, whereas G4-C bound to the lateral cell membranes demarcating the intercellular spaces. These findings suggest that colorectal G4 may be involved in crosslinking the lateral cell membranes of the surface-lining epithelial cells, thereby reinforcing epithelial integrity against mechanical stress exerted by the bowel lumen. (J Histochem Cytochem 47:75-82, 1999) PMID- 9857215 TI - Vesicular acetylcholine transporter in the rat cochlear nucleus: an immunohistochemical study. AB - After being synthesized in the cytoplasm of axon terminals, acetylcholine is packaged into synaptic vesicles by a proton-dependent transporter, vesicular acetylcholine transporter (VAChT). Localization of VAChT is restricted to cholinergic neurons, especially their terminals. We used an anti-VAChT antibody from INCSTAR to localize cholinergic terminals in the rat cochlear nucleus (CN), an important brainstem auditory center. VAChT immunoreactivity in the rat CN appears as labeled puncta and a few connecting fibers. In ventral CN (VCN), VAChT labeled puncta are closely associated with somatic profiles of medium to large neurons. In and near the granular regions of VCN, VAChT-labeled puncta are more diffusely scattered. In the subpeduncular corner and the medial sheet, some VAChT labeled fibers are seen in connection with especially prominent VAChT-labeled puncta. In dorsal CN (DCN), VAChT-labeled puncta show no clear association with somata and are found in all layers. Ultrastructurally, VAChT labeling is seen in the cytoplasm and is associated with synaptic vesicle membrane of terminals with small round vesicles. Such VAChT-labeled terminals synapse with cell bodies and dendrites in the CN.(J Histochem Cytochem 47:83-90, 1998) PMID- 9857216 TI - Exogenous 17beta-estradiol blocks alpha and mu but not pi class glutathione S transferase immunoreactivity in epithelium of Syrian hamster vas deferens. AB - Members of the glutathione S-transferase (GST) family of detoxification enzymes play a role in chemotherapy resistance in certain cancers but have not been directly implicated as agents whose absence may predispose tissues to hormonally induced tumorigenesis. Here we report the development of a polyclonal antiserum to a hamster mu class GST, and immunohistochemical analysis of alpha, mu, and pi class GSTs to study the effects of hormone treatment on their expression in reproductive tract tissues of male golden Syrian hamsters. These animals develop leiomyosarcomas in the vas deferens after treatment with testosterone propionate (TP) and 17beta-estradiol (E2). High levels of all three GST classes were detected throughout the reproductive tract epithelium of control animals. In 100% of the experimental animals, 4 weeks of treatment either with E2 alone, or with E2 plus TP promoted a complete loss of immunostaining for alpha and mu class GSTs, but not for pi class GSTs, only in the epithelial lining of the vas deferens. In contrast, treatment with TP alone resulted in moderate hyperplasia of smooth muscle in the proximal vas deferens, with no cellular atypia and no changes in immunoreactivity of any of the GST classes. The consistent and site specific nature of these results strongly suggests a functional role for GSTs in hormonally induced carcinogenic process. (J Histochem Cytochem 47:91-98, 1999) PMID- 9857217 TI - Ultrastructural localization of beta-actin and amphoterin mRNA in cultured cells: application of tyramide signal amplification and comparison of detection methods. AB - We describe a nonradioactive preembedding in situ hybridization protocol using digoxigenin-labeled RNA probes and tyramide signal amplification to increase the sensitivity of detection. The protocol is sensitive enough for electron microscopic localization of endogenous messenger RNAs encoding beta-actin and amphoterin. Three visualization methods were compared: diaminobenzidine enhanced by nickel, Nanogold enhanced by silver and gold toning, and fluorescently labeled tyramides. Diaminobenzidine and Nanogold can be used in both light and electron microscopy. The nickel-enhanced diaminobenzidine was the most sensitive visualization method. It is easy to accomplish but a drawback is poor spatial resolution, which restricts its use at high magnifications. Nanogold visualization has considerably better spatial resolution and is therefore recommended for electron microscopy. Fluorescent tyramides, especially TRITC tyramide, offer a good detection method for fluorescence and confocal microscopy. The methods were used to localize amphoterin and beta-actin mRNAs in motile cells. Both mRNAs were found in the soma and cell processes. In double labeling experiments, beta-actin mRNA localized to filamentous structures that also contained ribosomal proteins. Especially in the cortical cytoplasm, beta-actin mRNA was associated with actin filaments. Direct localization to microtubules was only rarely seen. (J Histochem Cytochem 47:99-112, 1999) PMID- 9857218 TI - Malignancies of the uterine corpus and immunoreactivity score of the DNA "mismatch-repair" enzyme human Mut-S-homologon-2. AB - We analyzed human Mut-S-Homologon-2 expression in normal endometrial tissue (n = 15) and malignancies of the uterine corpus (n = 40). Human Mut-S-Homologon-2 protein was investigated immunohistochemically on frozen sections, using a highly sensitive streptavidin-peroxidase technique and a specific mouse monoclonal antibody (clone FE11). Human Mut-S-Homologon-2 labeling pattern was compared with the staining pattern of the proliferation marker Ki-67 in the same tumors. A human Mut-S-Homologon-2 immunoreactivity score (human Mut-S-Homologon-2-IRS: negative 0-1; weak 2-3; moderate 4-6; strong 8-12) for semiquantitative analysis of human Mut-S-Homologon-2 expression is presented. In normal endometrial tissue samples we found weak nuclear immunoreactivity for human Mut-S-Homologon-2 in 67%, whereas the remaining 33% were negative for human Mut-S-Homologon-2 (mean human Mut-S-Homologon-2-IRS 1.25 +/- 1.29). All malignancies of the uterine corpus analyzed revealed moderate to strong nuclear immunoreactivity (mean human Mut-S-Homologon-2-IRS 9.00, +/- 3.16). Human Mut-S-Homologon-2 staining was heterogeneous, with visual differences among individual tumor cells. Expression of human Mut-S-Homologon-2 protein was consistently and strongly upregulated in tumor cells of malignancies of the uterine corpus compared with normal endometrial tissue (human Mut-S-Homologon-2-PP p<0.001; human Mut-S-Homologon-2 IS p<0.001; human Mut-S-Homologon-2-IRS p<0.001). No statistically significant correlation in comparing the labeling patterns for human Mut-S-Homologon-2 with the labeling patterns for Ki-67 (mean percentage of Ki-67-positive tumor cells 22.00% +/- 17.20) was observed in malignancies of the uterine corpus (human Mut-S Homologon-2-PP p=0.443; human Mut-S-Homologon-2-IS p=0.234; human Mut-S-Homologon 2-IRS p=0.173). Our findings indicate that human Mut-S-Homologon-2 is expressed in normal human endometrial tissue and that expression of human Mut-S-Homologon-2 may be of importance for the genetic stability of malignancies of the uterine corpus in vivo. (J Histochem Cytochem 47:113-118, 1999). PMID- 9857219 TI - Complete chromogen separation and analysis in double immunohistochemical stains using Photoshop-based image analysis. AB - Simultaneous detection of two different antigens on paraffin-embedded and frozen tissues can be accomplished by double immunohistochemistry. However, many double chromogen systems suffer from signal overlap, precluding definite signal quantification. To separate and quantitatively analyze the different chromogens, we imported images into a Macintosh computer using a CCD camera attached to a diagnostic microscope and used Photoshop software for the recognition, selection, and separation of colors. We show here that Photoshop-based image analysis allows complete separation of chromogens not only on the basis of their RGB spectral characteristics, but also on the basis of information concerning saturation, hue, and luminosity intrinsic to the digitized images. We demonstrate that Photoshop based image analysis provides superior results compared to color separation using bandpass filters. Quantification of the individual chromogens is then provided by Photoshop using the Histogram command, which supplies information on the luminosity (corresponding to gray levels of black-and-white images) and on the number of pixels as a measure of spatial distribution. (J Histochem Cytochem 47:119-125, 1999) PMID- 9857220 TI - Heat shock proteins: protective effect and potential therapeutic use (review). AB - The heat shock proteins (hsps) are induced by a variety of stressful stimuli and their overexpression has been shown to protect cells both in vitro and in vivo against such stimuli, as well as against stimuli-inducing apoptosis. The potential therapeutic benefit of elevating hsp levels in individuals with, for example, cerebral or cardiac ischaemia or neurodegenerative diseases has led to the identification of specific methods of inducing hsp expression in a non stressful manner. These include pharmacological procedures and cytokine treatment to elevate endogenous hsp levels and the development of viral vectors to deliver exogenous hsp genes. The advantages and disadvantages of each of these methods and their ultimate therapeutic potential are discussed. PMID- 9857221 TI - The riddle of L-glucose pentaacetate insulinotropic action (review). AB - The two anomers of L-glucose pentaacetate were recently found to stimulate insulin release. The insulinotropic action of these esters cannot be attributed to the catabolism in islet cells of their glucidic or acetic moieties. The present review deals with an alternative hypothesis. It is proposed that L glucose pentaacetate itself directly interacts with a yet unidentified receptor leading to plasma membrane depolarization, induction of electrical activity and increase in the cytosolic concentration of ionized Ca2+. This process displays analogies with the identification of bitter compounds by taste buds. Thus, beta-L glucose pentaacetate indeed displays a bitter, but no sweet, taste. Purified islet B-cells contain the alpha-gustducin G-protein involved in the perception of bitter taste by taste buds. The pentaacetate ester of beta-L-glucose decreases 86Rb outflow from prelabelled islets, provokes bioelectrical activity in islet B cells and may induce oscillations of cytosolic Ca2+ in these insulin-producing cells. The effects of beta-L-glucose pentaacetate upon glucagon and somatostatin secretion by the isolated perfused pancreas are also compatible with the present hypothesis. It is proposed that the L-glucose pentaacetate anomers could be used as novel insulinotropic tools in the treatment of non-insulin-dependent diabetes mellitus. PMID- 9857223 TI - Immunohistochemical expression of syndecan-1 in human endometrial cancer cells. AB - There are indications of increased frequency of endometrial cancer, one of the most common malignancies in women. Tissue samples of normal and malignant endometria were obtained post operatively from 30 women. We noted expression of syndecan-1 in 40% of investigated cancers. The most differentiated cancers showed 75% of positively stained specimens, moderately differentiated 40% and poorly differentiated neoplasm did not stain at all. In normal endometrial tissue syndecan-1 expression was regular and distinct in each specimen, but immunoreactivity of the hyperplastic endometrial specimens was absent. The detection of syndecan-1 in endometrial cancer of different clinical and histological stages could be of prognostic value in clinical diagnosis. PMID- 9857222 TI - Occurrence of acetaldehyde protein adducts formed in various organs of chronically ethanol fed rats: an immunohistochemical study. AB - We investigated the pathophysiological role of acetaldehyde protein adducts formed in vivo in organs of chronically alcohol fed male Wistar rats. Thirty male Wistar rats were fed on rodent pellets and 15% alcohol (V/V) for 5, 8 and 12 months, respectively before they were sacrificed. Further 30 male rats were chosen as the control group. We tested several organs by histological and immunohistochemical methods. Using immunohistological analysis, in the 12 months groups the basal membranes of glomerula, the membranes of liver, skeleton muscle and heart cells, and the gut were stained positively for acetaldehyde adducts. Using Western blotting of liver cell fractions (mitochondria/ lysosomes; microsomes; cytosol) adducts in charateristic molecular weight regions were detected. Approximately 30% of the sera of experimental rats contained antibodies against the acetaldehyde adducts formed in vivo. Immunologically detectable acetaldehyde adducts could be found in all rat organs tested. The stage of alcohol disease attained in this experiment after 12 months of ethanol feeding is described as the initial phase of manifestations of disturbances that are seen also in the carbohydrate metabolism. PMID- 9857224 TI - Radiation kills human peripheral T cells by a Fas-independent mechanism. AB - The mechanism by which radiation induces human peripheral T cell apoptosis is not known. We examined sequential changes in post-irradiated peripheral blood mononuclear cells (PBMC(S)) taken from normal volunteers, by using flow-cytometer and an anti-CD3 monoclonal antibody, annexin V, propidium iodide, anti-Fas antibody, and anti-Fas ligand antibody. After 5 or 10 Gy of irradiation with a 60Co radiation therapy unit, most of the human peripheral T cells showed positivity against annexin V in 15 h, and positivity against propidium iodide in 23 h after irradiation. On a microscopy-video system, approximately 80% of mononuclear cells revealed apoptotic changes in 24 h after irradiation. Because of its proposed role in activation-induced cytotoxicity, we also examined the Fas (CD95/Apo-1) pathway in killing T cells by irradiation. Irradiated PBMC, displayed no increase in surface Fas expression and caspase-3 activity relative to non-irradiated cells. In addition, the anti-Fas ligand failed to eliminate the apoptotic death of PBMC, after irradiation. These results suggest that irradiation induces direct apoptosis of T cells by a Fas-independent mechanism. PMID- 9857225 TI - Endopeptidase 24.11/CD10 is down-regulated in renal cell cancer. AB - The regulatory mechanisms responsible for malignant transformation, tumor progression and metastasis in renal cell cancer (RCC) are still unclear, but there is some evidence that biologically active peptides might have regulatory effects on the behavior of this malignancy. Tumor cells can change local concentrations of active peptides by modulating their cell-surface enzymes. Using immunohistochemistry and enzyme-histochemistry, the expression of various membrane peptidases was examined in RCC and adjacent noninvaded renal parenchyma (n = 44). We describe the down-regulation of neutral endopeptidase 24.11 (NEP) protein expression in RCC of the clear cell/chromophilic type when compared with renal parenchyma, and show for the first time the lack of enzyme activity of NEP in RCC. The strongest expression could be found for dipeptidyl peptidase IV (DPIV) which is only decreased in RCC of the chromophobe cell type and is even present in oncocytoma. Aminopeptidase N (APN) and aminopeptidase A (APA) show attenuated expression in up to one third of clear cell/ chromophilic RCC. Chromophobe RCC and oncocytomas do not express APN, APA, NEP and gamma glutamyltranspeptidase. PMID- 9857226 TI - A fast non-fluorescent staining method for the detection of primed in situ synthesis products. AB - For the assessment of chromosomal numeral aberrations in cells, the method of choice is fluorescent in situ hybridisation (FISH) or, a newly introduced technique, oligonucleotide primed in situ hybridisation (PRINS). In the PRINS method labeled nucleotides are incorporated into newly synthesized DNA mediated through the Taq polymerase. Both PRINS and FISH reactions are visualized with a fluorescent light detection system. We present a method whereby PRINS products can be reliably and rapidly visualized by a streptavidin DAB detection system. PMID- 9857227 TI - A study on the relationship between antisense EGFR cDNA fragments and nuclear matrix proteins in glioblastoma cells. AB - The association of antisense epidermal growth factor receptor (EGFR) cDNA fragments with nuclear matrix from EGFR-antisense transfected glioblastoma cell lines U343 and U87 was investigated. A 1015 bp DNA fragment (primer I-II) was amplified in both genomic DNA and nuclear matrix-associated DNA (NM DNA) from EGFR-antisense transfected glioblastoma cell lines U343E and U87E. Two different DNA fragments (940 bp and 110 bp) were amplified by primer I-III in both genomic DNA and NM DNA of U343E, while one 110 bp PCR product was shown with the same primer in both genomic DNA and NM DNA of U87E only. After EGFR-antisense transfection, the binding property of the 110 bp DNA fragment (primer IV-V) to nuclear matrix was not affected. Southwestern blotting demonstrated the presence of antisense EGFR cDNA binding nuclear matrix proteins. Our findings demonstrate that not only EGFR DNA is associated with nuclear matrix, but the transfected antisense EGFR cDNA also binds to nuclear matrix proteins. The nuclear matrix is most likely involved in the replication and transcription of antisense EGFR cDNA or hybridisation with sense mRNA in vitro. PMID- 9857228 TI - Enhancement of a constitutively active promoter for gene therapy by a positive feed-back transcriptional activator mechanism. AB - Success of gene replacement therapy depends on long-term, high level expression of the transgene. Gene therapy vectors incorporating a promoter of a constitutively active eukaryotic gene may allow long-term expression in vivo, but the expression level may be insufficient for therapeutic effects. To enhance transcription from eukaryotic promoters, a strategy with dicistronic vectors encoding the therapeutic gene of interest together with a transcription factor that binds and activates the promoter was tested. Expression vectors for the chimeric tet repressor/VP16 transcription factor (tTA) driven by the human beta actin promoter were constructed, and tandem tet operators were inserted within the promoter. This arrangement significantly enhanced expression of G-CSF in fibroblasts to higher levels than the immediate/early CMV promoter. Stably transfected fibroblast clones produced up to 2.4 microg G-CSF per 10(6) cells x 24 h. After injection of genetically engineered cells into SCID mice, the enhanced beta-actin promoter construct resulted in marked leukocytosis, whereas the unmodified promoter had only a marginal therapeutic effect. Transcription factor-enhanced, feed-back-activated human promoters may thus achieve higher expression levels than viral control elements, and may be advantageous for gene therapy due to high constitutive activity in vivo. PMID- 9857229 TI - Functional roles of HIV accessory proteins for viral replication (review). AB - Numerous lentiviruses, including human immunodeficiency virus types 1 and 2 (HIV 1 and HIV-2) of causative agents of human AIDS as representative, have been recently isolated from various species of primates. The fundamental and most prominent feature of the viruses is the presence of a number of accessory genes in their genomes. Extensive biological and biochemical studies have demonstrated that the accessory gene products are not essential for viral replication at least in certain types of cells. Quite surprisingly, some of these accessory proteins are absolutely non-essential in any types of cells so far examined. In this brief review, our systematic genetic studies on the importance of the accessory proteins of HIV-1 and HIV-2 for viral replication are described and discussed. PMID- 9857230 TI - Cytotoxicity of Fas ligand against lymphoma cells with radiation-induced Fas antigen. AB - Fas antigen, also termed APO-1 or CD95, is a transmembrane protein and a member of the tumor necrosis factor receptor/nerve growth factor receptor superfamily which mediates apoptosis upon oligomerization. The Fas/Fas ligand system is considered to be a key regulator of apoptosis. Recently, we have demonstrated that Fas antigen expression is induced by low-dose irradiation of some types of lymphomas, and we also demonstrated that irradiation-induced Fas antigen expression increased with the passage of time until peaking at 48 h after irradiation in CML-C1, CML-C2, DL-40, and DL-95 cell lines. In this study, we also examined the potential cytotoxicity of Fas ligand peptide against several types of lymphoma/leukemia cell lines that showed induction of Fas antigen expression under irradiation. Flow cytometry analysis was performed at 6, 24 and 48 h after irradiation. Samples (1 x10(6) cells/ml) from irradiated and non irradiated cells of each cell line were incubated with or without 5 microg/ml of Fas ligand peptide for 2 h at 37 degrees C in a humidified atmosphere of 5% carbon dioxide (CO2) in air. The killing effect of Fas ligand against cell lines of CML-C1, DL-40, and DL-95 were clearly identified as the percentage of cells with Fas antigen expression induced by irradiation. Concerning HD-70 cell line, for which soluble Fas antigen has been identified, the killing effects were clearly observed in samples pre-treated with PBS washings. To our knowledge, this is the first report describing a possible application of the Fas/Fas ligand system in treatment of certain types of malignancies in which Fas antigen is inducible by irradiation. PMID- 9857231 TI - Telomere length, telomerase activity and telomerase RNA expression during mouse mammary tumor progression. AB - To investigate the roles of telomere length (mean length of the terminal restriction fragments; TRFs), telomerase activity (TA) and telomerase RNA (mTR) expression in relation to mouse mammary tumor progression, we examined a pregnancy-dependent mouse mammary tumor line (TPDMT-4) and its four autonomous sublines (T4-OI320: non-metastatic; and T4-OI165, -OI96, and -OI145: artificial metastatic) of DDD/1 mouse origin, and an autonomous growing mammary tumor (JYG MC) showing spontaneous lung metastasis developed in BALB/c mice infected with a Chinese feral mice (Sub-Jyg)-derived mouse mammary tumor virus (JYG-MTV). Compared with normal (pregnant) mammary tissue, the TA was elevated in the TPDMT 4 tumor and in the non-metastatic subline tumor (T4-OI320) (x10 fold, respectively), and was further increased (x13-15 fold) in parallel with the acquisition of metastatic potential (T4-OI165, -OI96, and -OI145). The mTR level was upregulated (x2.7-2.8 fold) in all autonomous growing tumors compared to the normal counter-part, but not in TPDMT-4. The TRF was shorter in accord with tumor progression (normal mammary tissue, 48 kb; TPDMT-4, 45 kb; T4-OI320, 37 kb; T4 OI165, -OI96 and -OI145, mean 37.7 kb; and JYG-MC, 21 kb). These results suggest that the activation of TA occurs as an early event at the stage of hormone dependent tumorigenesis, followed by the up-regulation of mTR expression in accordance with the acquisition of autonomous growth, and then further activation of TA occurs when the tumor acquires metastatic potential. The TRF shortening was in parallel with the tumor progression. PMID- 9857232 TI - Germinated barley foodstuff increases fecal volume and butyrate production at relatively low doses and relieves constipation in humans. AB - Germinated barley foodstuff (GBF), derived from the aleurone layer, scutellum and germ of germinated barley, contains a large quantity of fermentable dietary fibers, especially hemicellulose. GBF was given to 9 healthy volunteers in a dose of 9 g of GBF per day for 10 consecutive days, and subsequently 18 g of GBF for another 10 days. As a control, no GBF was given for 3 days before administration of GBF (control period). Fecal weight, water content and short chain fatty acid content were measured before and during the last 3 days of each experimental period. Feeding of GBF significantly increased the fecal butyrate content as well as fecal weight at both dose-levels (9 and 18 g/day), compared with those during the control period. Daily administration of 9 g GBF induced the maximum level of defecation in humans. Relatively mild but chronic constipated volunteers (n = 16) were administered 9 g of GBF daily for 14 days. In this experiment, the condition of defecation (frequency, volume) was estimated by a questionnaire survey. GBF significantly improved defecation within a short period without severe adverse effects. No major abnormalities in laboratory findings were found in hematologic and urinary analyses. In conclusion, daily administration of 9 g GBF was effective for improving defecation in healthy but constipated humans. GBF is a highly safe and effective foodstuff for improving defecation. PMID- 9857233 TI - Quantification of Ggamma- and Agamma-globins by electrospray ionisation mass spectrometry. AB - Elucidation of the molecular basis for persistent fetal haemoglobin (Hb F) production in adult life has important implications for the pathophysiology and treatment of human beta haemoglobinopathies. Electrospray ionisation mass spectrometry (ESMS) was applied to analyse the pattern of gamma-globin expression in patients with hereditary persistence of fetal haemoglobin (HPFH) and sickle cell anaemia (SCA). Ggamma and Agamma-globin chains were identified by their measured molecular masses and distinguished by mass difference (14 Da) following deconvolution of ESMS spectra using maximum entropy based software. Prediction of HPFH type by ESMS was confirmed by molecular analysis. Direct determination of Ggamma:Agamma globin chain ratio from whole blood by the novel application of ESMS provides a rapid and sensitive approach to characterisation of gamma-globins and facilitates correlation of gamma-globin level and polymorphism of cis-active elements at the beta-globin locus. PMID- 9857234 TI - Establishment of a quantitative RT-pCR for detection of vascular cell adhesion molecule-1 transcripts in endothelial cells after stimulation with advanced glycation endproducts. AB - Advanced glycation endproducts (AGE) are supposed to increase endothelial expression of adhesion molecules like vascular cell adhesion molecule-1 (VCAM-1) by inducing an intracellular stress with subsequent activation of nuclear transcription factor NF-kappa-B. Quantitative analysis of VCAM-1-transcription has not been demonstrated concerning this topic. Thus, the aim of this study was to establish quantitative reverse transcription polymerase chain reaction (RT PCR) assays using a spacer gene in order to measure the amounts of specific mRNA for VCAM-1 in human umbilical vein endothelial cells (HUVEC) which were stimulated with AGE-albumin (AGE-BSA). A recombinant RNA-standard was synthesized and used as internal RT-PCR standard. The amount of VCAM-1-mRNA in unstimulated HUVEC was found to be 2.2 +/- 2.7 copies per cell. After stimulation with AGE BSA, mRNA-levels were elevated to 38.9 +/- 10.9 copies per cell. Positive controls (stimulated with lipopolysaccharide) revealed mRNA-levels of 78.7 +/- 27.5 copies per cell. We conclude that quantitative RT-PCR using the spacer gene technique is a valid and reliable method for the measurement of small amounts of specific PMID- 9857236 TI - Menstrual state should be considered in determining sero-positivity of soluble angiogenic factors in breast cancer. AB - We investigated whether blood angiogenic factor (vascular endothelial growth factor, VEGF; angiogenin; basic fibroblast growth factor, bFGF; platelet-derived growth factor-AB, PDGF-AB) levels change during menstrual cycle of healthy premenopausal females or after menopause. We also measured the serum angiogenic factor levels in 34 operable breast cancer patients and compared them to those of healthy volunteer controls. No differences in the four angiogenic factor levels were found between the follicular and luteal phases of normal menstruation. However, angiogenin and bFGF levels were higher in pre-menopausal females than post-menopausal female and young male healthy volunteers. In cancer patients, the sero-positivity rate of the bFGF was 8.8% with menstrual-state-unmatched cut-off points, which increased to 36.4% with menstrual-state-matched cut-off points. This discrepancy was especially high in post-menopausal cancer patients. In conclusion, physiological elevation of the bFGF during normal menstruation can influence the precise interpretation of the pathological elevation of the bFGF in pre-menopausal breast cancer patients. PMID- 9857235 TI - Thymidine phosphorylase activity in tumor correlates with venous invasion. AB - Thymidine phosphorylase (dThdPase) is identical to platelet-derived endothelial cell growth factor (PD-ECGF), an angiogenesis factor. We investigated the correlation between dThdPase activity in gastric cancer tissue and clinicopathological factors. Thirty-three cancer tissue specimens and 23 adjacent normal gastric mucosal specimens were obtained from surgery. Measurement of dThdPase activity was based on the amount of 5-fluorouracil formed from 5'-deoxy 5-fluorouridine by dThdPase. Mean dThdPase activity in cancer tissue was approximately 3.2 times higher than that in normal tissue. Cancerous tissues with venous invasion had about twice the dThdPase activity as cancerous tissues without venous invasion. Other clinicopathological features were not related to dThdPase activity. A correlation between dThdPase activity and immunosuppressive acidic protein level was observed (r = 0.532, P = 0.005). dThdPase activity in gastric cancer cells was found to be correlated with venous invasion, supporting previous findings that it plays a role in tumor angiogenesis. PMID- 9857237 TI - Enhancement of in vitro and in vivo anti-tumor activity of anti-GD2 monoclonal antibody 220-51 against human neuroblastoma by granulocyte-macrophage colony stimulating factor and granulocyte colony-stimulating factor. AB - We have evaluated the anti-tumor effect of anti-GD2 mouse monoclonal antibody (mAb) 220-51 against human neuroblastoma cell line TGW in vitro and in vivo. The mAb 220-51 was able to mediate complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) using human effector cells. In the presence of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte ADCC was significantly augmented in vitro. When mAb 220-51 was administered to tumor-bearing nude mice, tumor growth was significantly inhibited as compared with untreated controls. Administration of recombinant murine GM-CSF in combination with mAb 220-51 significantly enhanced the anti-tumor effect of mAb in vivo. Recombinant human granulocyte colony stimulating factor (G-CSF) combined with mAb 220-51 was also able to enhance it, although granulocyte ADCC was not affected by the presence of recombinant human G CSF in vitro. Moreover, GM-CSF and G-CSF work additively to enhance the anti tumor effect of mAb 220-51 in vivo. The GM-CSF and G-CSF may have a clinical potency in immunotherapy with anti-GD2 mAb for the treatment of neuroblastoma. PMID- 9857238 TI - Possible prediction of adverse reactions to fluorouracil by the measurement of urinary dihydrothymine and thymine. AB - Dihydropyrimidine dehydrogenase (DPD) deficiency with a defect of the pyrimidine catabolic pathway has recently become the focus of considerable attention, due to the severe 5-fluorouracil (5-FU) toxicities occurring in DPD deficiency patients. Studies also suggest that 5-FU toxicities could occur in another pyrimidine metabolic disorder, dihydropyrimidinuria (DHPuria). This study shows that urinary dihydrothymine (DHT) and thymine (THY) are useful indexes for detection of DPD deficiency and DHPuria. We measured urinary DHT and THY in 276 Japanese adults to establish reference ranges. When males and females were compared, both DHT and THY levels were found to be significantly higher in females. The reference ranges (mean +/- SD with logarithmic values) for males were found to be 1.56-5.70 micromol/g of creatinine for DHT and 0.40-1.47 micromol/g of creatinine for THY. The reference ranges for females were found to be 1.89-8.33 micromol/g of creatinine for DHT and 0.58-2.30 micromol/g of creatinine for THY. In addition to this study we analyzed a DPD deficiency case and a DHPuria case. In the DPD deficiency case, the THY concentrations of all urine samples were out of the reference range. However, uracil levels in most of the samples were within the normal range. The DHPuria case excreted large amounts of DHT and dihydrouracil, both out of the normal range. PMID- 9857239 TI - Insulin-like growth factor-1 and insulin-like growth factor receptor in thyroid tissues of patients with Graves' disease. AB - Growth factors are frequently involved in the regulation of mitosis and differentiation of several cell types and insulin-like growth factor-1 (IGF-1) is actively involved in the thyroid stimulating hormone-mediated proliferation of thyrocytes. In view of the pivotal role of IGF-1 in thyrocyte proliferation and of the still unsettled role of this growth factor in the pathogenesis of hyperplastic thyroid lesions, we investigated the expression of IGF-1 and of its corresponding receptor, by means of immunohistochemistry, in the surgical specimens obtained from six patients with Graves' disease. Moreover, IGF-1 mRNA expression was analysed in one such case by means of Northern hybridisation. All samples showed consistent intracytoplasmic immunoreactivity for both IGF-1 and IGF-1 receptor; the vast majority of hyperplastic thyrocytes were strongly decorated by the two antibodies used in this study whereas stromal cells displayed IGF-1 immunoreactivity only. IGF-1 mRNA was markedly overexpressed in Graves' disease in comparison with normal thyroid tissues. The results of this study suggest that IGF-1 and IGF-1 receptor may be actively involved in the pathogenesis of Graves' disease; furthermore, IGF-1 and IGF-1 receptor apparently act by different mechanisms (paracrine vs. autocrine) as suggested by their differential expression in epithelial and stromal cells. PMID- 9857240 TI - Regulation of human prostate-specific antigen gene expression in transgenic mice: evidence for an enhancer between the PSA and human glandular kallikrein-1 genes. AB - The human prostate-specific antigen (PSA) and glandular kallikrein-1 (hGK-1, also known as hK2) genes are tandemly located on chromosome 19, separated by a 12-kb intergenic region. The coordinate regulation of these two genes suggests the presence of common regulatory elements responsible for tissue specificity and/or levels of expression within this region. To identify such regulatory elements, we generated two sets of transgenic mice, which had incorporated either the PSA gene alone or together with the intergenic region. Both sets of transgenics exhibit remarkably prostate-specific expression of the transgene. However, the presence of the intergenic region abrogates the dependence on high PSA gene copy-number for high levels of PSA expression. This suggests the existence of a positive regulatory element in the intergenic region. By using a previously identified distal enhancer element of PSA (termed DEE 1) as a probe, we identified a cross hybridizing fragment, which we termed DEE 2, in the intergenic region. Sequence analysis shows that DEE 2 is 76% identical to DEE 1, and it includes a putative androgen-responsive element. Here, we propose a model to illustrate how the two enhancers may work to regulate the transcription of PSA and hK2. PMID- 9857241 TI - Measurement of circulating immunoreactive alpha-atrial natriuretic peptide in human plasma: validation and clinical application. AB - Recent evidence suggests that plasma alpha-atrial natriuretic peptide (alpha-ANP) may serve as a useful biochemical marker of severe heart failure, as circulating levels become ighly elevated. In the present study, a specific radioimmunoassay (RIA) protocol was adapted and optimised for rapid quantitative measurement of circulating alpha-ANP levels in plasma taken from clinically normal subjects and heart transplant recipients with no evidence of heart failure or heart transplant rejection. The plasma concentration of immunoreactive alpha-ANP in heart transplant recipients (115 +/- 10 pg/ml, n = 14) was significantly higher (p < 0.01) than in normal subjects (14 +/- 5 pg/ml, n = 20). Comparison of circulating plasma immunoreactive alpha-ANP levels obtained by the adapted assay to levels obtained by a standard RIA protocol revealed a significant correlation (r = 0.998, y = 1.004 x -2.94, n = 40). The adapted assay is simple, precise, and with a shorter incubation time that would enable results to be available within one working day. These results show that circulating biological active a-ANP levels are elevated in healthy heart transplant recipients with no evidence of heart failure; thus extending previous reports concerning the elevation in circulating alpha-ANP levels from patients with a wide range of heart disorders. The adapted RIA protocol would facilitate rapid detection and quantification of alpha-ANP in experimental research and routine clinical investigations. PMID- 9857242 TI - Histochemically demonstrable acid phosphotyrosine phosphatase activity in human tissues. AB - A histochemical method for the demonstration of activity of acid phosphatase(s) hydrolyzing o-phospho-L-tyrosine at pH 6.2 is described. Due to the instability of ordinary acid phosphatase incubation media containing phosphotyrosine and lead salt, the latter was dissolved in citrate solution (possibly forming a chelate) to obtain a stable incubation mixture. Localization of enzyme activity at the cellular level was good. Phosphotyrosine is known to be hydrolyzed at slightly acid pH by prostatic, macrophagic, and low-molecular weight cytoplasmic acid phosphatases and phosphotyrosyl protein phosphatases. These enzymes have been suggested or are established to be involved in cellular regulation by dephosphorylation of tyrosyl residues of proteins formed by various protein tyrosine kinases. An unexpected finding from various tissues was that the widely distributed lysosomal acid phosphatase did not contribute to the histochemical reaction, possibly due to poor hydrolysis of phosphotyrosine by lysosomal-type acid phosphatase. Various human tissues were studied for their acid phosphotyrosine phosphatase activities, which were then quantitated photometrically. Highest activity was found in the prostate glands, seminal vesicle epithelium, endometrial secretory glands, alveolar macrophages, stratum granulosum of the skin, and lymph nodes. The contribution of various enzymes hydrolyzing phosphotyrosine to histochemically detected activity in different tissues is discussed here. PMID- 9857243 TI - Fibronectins and basal lamina molecules expression in human subcutaneous white adipose tissue. AB - Adipose tissue is a type of connective tissue whose extracellular matrix (ECM) components are poorly characterized in vivo. Several in vitro studies have suggested a regulatory role for the ECM molecules during adipocyte differentiation. Since no data are available concerning the in vivo expression of the main ECM components such as fibronectin, collagen IV, laminin and heparan sulfate proteoglycan in the adipose tissue, we investigated the presence of these molecules by immunohistochemistry. We show that fibronectin isoforms are not expressed in fully differentiated subcutaneous adipocytes whereas collagen IV, laminin and heparan sulfate are detectable around single adipocytes, i.e. in the region corresponding to the basement membrane. These data are supported by previous in vitro studies showing a strong decrease of fibronectin synthesis during adipocyte development whereas basement membrane molecules seem to increase during adipocyte differentiation. PMID- 9857244 TI - Immunohistochemical study of tonsils from newborn infants with emphasis on follicular dendritic reticulum cells. AB - The origin of follicular dendritic reticulum cells (FDRCs) is still debated in the literature, although their derivation from local transformed mesenchymal cells is now generally accepted. The purpose of this immunohistochemical study was to further define the nature of the FDRC, for which an endothelial cell derivation was proposed in previous papers. Palatine tonsils were removed at autopsy from eight newborn infants ranging from a few hours to four days of age. Paraffin embedded tonsil specimens were stained by current histochemical methods and immunohistochemically processed with a panel of antibodies against follicular dendritic reticulum cells (CD21 and CD35), endothelial cells (Factor VIII R. Ag, CD31, CD34), B lymphoid cells (L26, LN1, MB2), T lymphoid cells (UCHL1, DF-T1, OPD4, CD8, CD3), monocytic cells (PGM1 and KP1), interdigitating dendritic cells (S100 Protein, PGM1), intercellular adhesion molecule (ICAM-1 (CD54). Double immunostainings for endothelial markers and for FDRCs were performed. In newborn infant tonsils, lymphoid follicles are absent. T lymphocytes appear to represent the largest component whereas B lymphocytes are seen in small aggregates between blood capillary vessels. Monocytes and interdigitating dendritic cells are also present. The capillary endothelial cells within the B lymphoid aggregates are positive for endothelial and FDRCs markers; double immunostaining for Factor VIII R Ag (or other endothelial markers) and CD21 was present within the same capillary endothelial cells. Further they were positive for ICAM-1. These observations lend further evidence that the derivation of FDRCs may be from transformed endothelial cells and on this way they act as enhancing microenvironment for B lymphocyte expansion. PMID- 9857245 TI - Immunohistochemical study of Tn, sialosyl-Tn and T antigens in human meningiomas. AB - Since an aberrant glycosylation associated with abnormal expression of cell surface carbohydrates has been described in many human tumours, we have investigated the expression of simple mucintype carbohydrate antigens (Tn, sialosyl-Tn and T) in 30 formalin-fixed paraffin-embedded surgical specimens of primary meningiomas, subdivided according to the WHO classification as follows: 18 meningotheliomatous, 7 transitional, 2 angiomatous and 3 anaplastic; in addition, 5 meningeal samples taken at autopsy were tested as control. Specific monoclonal antibodies (HB-Tn1, HB-STn1 and HB-T1, Dako) were utilized with ABC method; the percentage of immunostained cells was graded as follows: O (no staining); 1 (> 0 to 5%); 2 (> 5 to 50%); 3 (> 50%). Tn antigen was not expressed in any case. Sialosyl-Tn antigen was demonstrated only in cases of anaplastic meningiomas (score 1-3). The immunoreactivity for T antigen was found in 2 cases of meningotheliomatous (score 1) and in all the anaplastic meningiomas (score 1 2). Negative results were obtained with antisera in the control meningeal samples. Our results thus document a peculiar expression of sialosyl-Tn and T antigens in anaplastic meningiomas, suggesting a block of glycosylation as an immunophenotypic finding of malignant transformation. The sporadic immunostaining for T antigen encountered in the meningotheliomatous variety may be interpreted as an intermediate step of malignant changes. PMID- 9857246 TI - Application of fluorescence resonance energy transfer techniques to the study of lectin-binding site distribution on Paramecium primaurelia (Protista, Ciliophora) cell surface. AB - Fluorescence resonance energy transfer (FRET) is a photophysical phenomenon occurring between the molecules of two fluorochromes with suitable spectral characteristics (donor-acceptor dye pair), and consisting in an excitation energy migration through a non-radiative process. Since the efficiency of the process is strictly dependent on the distance and reciprocal orientation of the donor and acceptor molecules, FRET-based techniques can be successfully applied to the study of biomolecules and cell component organisation and distribution. These techniques have been employed in studying Paramecium primaurelia surface membrane for the reciprocal distribution of N-acetylneuraminic acid (NeuAc) and N acetylglucosamine (GlcNAc) glycosidic residues, which were found to be involved in mating cell pairing. NeuAc and GlcNAc were detected by their specific binding lectins, Limulus polyphemus agglutinin (LPA) and wheat germ agglutinin (WGA), respectively. Microspectrofluorometric analysis afforded the choice of fluorescein isothiocyanate and Texas red conjugated with LPA and WGA, respectively, as a suitable donor-acceptor couple efficiently activating FRET processes. Studies performed both in solution and in cells allowed to define the experimental conditions favourable for a FRET analysis. The comparative study carried out both on the conjugating-region and the non conjugating region of the surface membrane, indicates that FRET distribution appears quite homogeneous in mating-competent mating type (mt) I, whereas, in mating-competent mt II cells, FRET distribution seems to be preferentially localised on the conjugating-region functionally involved in mating cell pairing. This difference in the distribution of lectin-binding sites is suggested to be related to mating-competence acquisition. PMID- 9857247 TI - Distribution of neurofilaments in the telencephalon and mesencephalon of the adult and developing Gallotia galloti lizard. AB - The location and chronology during development of the immunoreactivity due to the presence of neurofilaments (NF) in telencephalon and mesencephalon of the lizard Gallotia galloti has been studied. For this purpose we have used two antibodies recognizing both phosphorylated and non phosphorylated neurofilaments (NF), a polyclonal Ab (NF 005), and a commercial monoclonal antibody (NF-200). The study was completed by using the Bielschowsky technique. During ontogeny, the anti-NF 005 immunoreactivity appeared at E40 in some tracts in mesencephalon and increased in intensity in isolated nerve fibers, tracts and commissurae till adult. However, a weak staining appeared in some neurons. In telencephalon, the reactivity was detected only in adult specimens. It was clearly more abundant in mesencephalon than in telencephalon, which could indicate that a greater complexity and functional importance exist in the lizard midbrain in relation to other primitive regions as the basal nuclei and cortical areas. In contrast to young specimens, the monoclonal anti-NF 200 was detected in neuronal perikarya, dendrites and axons in adults. Thus, in lizards, both antibodies highly recognized phosphorylated and non-phosphorylated forms of proteins of NF (NF-H). In mammals, these forms of proteins are implicated in axonal maturation. The presence of these NF in reptiles, identified for the first time, proved to be phylogenetically stable. The anti-NF immunoreactivity distribution occurs both caudo-rostrally and from the ventral to the dorsal regions. PMID- 9857248 TI - Ultrastructure of enkephalin- and substance P-immunoreactive axon terminals in the hypoglossal nucleus of the chicken. AB - The morphology and distribution of leucine-enkephalin (LENK)- and substance P (SP)-containing axon terminals in the hypoglossal nucleus of the chicken was investigated immunocytochemically at the electron microscopic level. The major findings of the present study were as follows: 1) Both LENK- and SP immunoreactive (IR) profiles including unmyelinated axons, varicosities and axon terminals were found throughout the hypoglossal nucleus. 2) LENK-IR profiles without synapses (preterminal axons and varicosities) were observed more frequently (66%) than LENK-IR synaptic profiles (34%). In contrast, SP-IR synaptic terminals were seen more often (68%) than SP-IR terminals without synapses (32%). 3) Both LENK- and SP-IR synaptic terminals ending on dendrites were more numerous (83% and 94%, respectively) than those making synapses on somata (17% and 6%, respectively). 4) The average diameter of SP-IR synaptic terminals were significantly larger than that of LENK-IR ones. 5) Both LENK- and SP-IR synaptic terminals contained numerous small clear vesicles and several large dense-cored vesicles (LDV). Although both LENK- and SP-IR synaptic terminals contained LDV consistently, SP-IR synaptic terminals contained significantly more LDV than LENK-IR types. 6) Postsynaptic dense bodies (Taxi bodies) were seen more frequently in SP-IR synaptic terminals than in LENK-IR ones. These findings confirm both LENK and SP innervation in the hypoglossal nucleus and suggest a functional role as neuromodulator for the two neuropeptides in tongue control. PMID- 9857249 TI - Microscopical localization of actin-associated proteins alpha-actinin, tropomyosin and vinculin in motile human neutrophils. PMID- 9857250 TI - [Quality assurance for rational diagnosis and therapy in dermatology]. AB - Today quality is not only in the economy but also in medicine a prime parameter in the evaluation of goods and services. To guarantee quality in a comprehensive way not only quality control but also quality assurance or even quality management are needed. One has to focus on its integral aspects which are structure, process and result. With laboratory analysis a preanalytic phase has to be distinguished from an analytic and a postanalytic one. According to the notion of the Deutscher Arztetag quality assurance is a common task of all physicians. A variety of organisations provide support. Guidelines of various types are pivotal. Some concerning dermatology have already been released by Deutsche Dermatologische Gesellschaft; others are in the process of formulation. A comprehensive monograph is under way. PMID- 9857251 TI - [MIA ("melanoma inhibitory activity"). Biological functions and clinical relevance in malignant melanoma]. AB - The protein MIA was identified and isolated from the tissue culture supernatant of melanoma cells in vitro by its ability to inhibit thymidine incorporation by melanoma cell lines. After purification and partial sequencing of the peptide, a fragment of the MIA cDNA was cloned by RT-PCR. This cDNA fragment was used to screen phage libraries and subsequently fully encoding human and murine MIA cDNA and genomic DNA clones were obtained. The MIA gene spans a region of approximately 2 kb and is divided into 4 exons. Mapping the MIA gene revealed that the human gene is located on chromosome 19 and the murine gene on chromosome 7. The MIA open reading frame spans 131 (human) or 130 (murine) amino acids. The first 24 (human) or 23 (murine) amino acids represent a signal sequence directing the secretion of MIA into the extracellular compartment. The mature, secreted MIA consists of 107 amino acids and its MW is approximately 11 kDa. Preliminary structural data suggests that MIA is a small globular protein stabilized by two intramolecular disulfide bonds. Expression studies of protein und mRNA levels indicate that MIA is expressed specifically by malignant melanoma cells and chondrocytes. This points to a highly restricted expression pattern which is controlled by the MIA promoter. In addition, MIA provides a clinically useful parameter in patients with malignant melanoma. Enhanced values were measured in the serum of all patients with metastatic melanoma (stage III and IV). In vitro and in vivo experiments using recombinant MIA protein revealed that MIA specifically inhibits attachment of melanoma cells to fibronectin and laminin. Further analysis indicated a direct binding between MIA and the matrix proteins. This finding provides an explanation for the capability of MIA to inhibit proliferation of melanoma cells in vitro. Our studies suggest a putative function of MIA in regulated detachment of melanoma cells from the extracellular matrix which is an important step in metastasis. PMID- 9857252 TI - [Immunochemotherapy of metastatic uveal melanoma with interferon alfa-2b, interleukin-2 and fotemustine. Case reports and review of the literature]. AB - In spite of their tumor's origin in the uveal tract, many patients suffering from advanced uveal melanoma are admitted to dermatological oncology units. Most patients with metastases from uveal melanoma receive treatments that were established for stage IV cutaneous melanoma. However, both the biology as well as the metastastic behaviour of this tumor is different from cutaneous melanoma. Lymphatic metastases do not occur, and hematogeneous metastases usually occur later and predominantly involve the liver. The prognosis is very bad ranging from 2 to 5 months. We describe three patients with advanced uveal melanoma who received immunochemotherapy containing interferon-alpha 2b, interleukin-2, and fotemustine. This therapy induced a partial response of more than 49 months duration in one patient, whereas for the remaining patients the disease progression could be stabilized for eight and 16 months, respectively. This therapeutic success is reflected by a prolonged survival of 14,43+, and 59+ months. PMID- 9857253 TI - [Adjuvant therapy of pemphigus vulgaris and pemphigus foliaceus with intravenous immunoglobulins]. AB - High dose intravenous immunoglobulins (IVIG) have been shown to be effective in different autoimmune diseases. We report on the use of IVIG in patients with pemphigus vulgaris (PV) or pemphigus foliaceus (PF) resistant to conventional therapy. Six patients who presented with a relapse of their disease following conventional immunosuppressive treatment with prednisolone and azathioprine were additionally treated with 2 g/kg of IVIG every four weeks. Six to nine cycles of this therapy were given to each patient. All patients showed healing of their blisters without evidence of relapse even when steroids and immunosuppressants were tapered. None of the patients relapsed within one year of follow-up, although IVIG treatment was discontinued. Side effects of IVIG treatment were moderate with only slight evidence of headaches. We suggest that adjuvant treatment of PV and PF resistant to conventional immunosuppressive strategies is a useful addition to our therapeutic arsenal in both diseases. PMID- 9857254 TI - [Epidemiology of sexually transmitted diseases in the catchment area of the Rostock University Dermatology Clinic after 1945]. AB - The decline of STD in the region of Rostock, Germany, since 1945 is explained by a decreasing incidence of gonorrhoea and trichomoniasis. On the other hand chlamydial and candidal infections have increased. The rate of chlamydia infections was about 45% in our venerological patients, as common as gonorrhea used to be. Genital candidal infections were to be found in 23%. Mycoplasmas were identified in 15%, usually combined with other pathogens except in some cases of vaginitis. PMID- 9857255 TI - [Pustular dermatitis after administration of a too highly concentrated dithranol ointment in seborrhea]. AB - Seborrhoic dermatitis is an erythematosquamous condition of unknown etiology with a prevalence of approximately 2.5%. Frequently difficult to treat, it may respond to the application of low doses of dithranol, a substance which can induce a pustular dermatitis as an adverse side effect depending on the applied concentration and individual susceptibility. We describe a pustular dermatitis after the application of a preparation containing dithranol at an erroneously high concentration in a 30-year-old patient with seborrhoic dermatitis. PMID- 9857256 TI - [Papillary cystadenoma of the minor salivary glands]. AB - Papillary cystadenoma of the minor salivary glands is a rare benign tumor that clinically resembles a mucous cyst. We demonstrate its histologic features and differential diagnosis by a case report. Benign and malignant neoplasms of the minor salivary glands are not well acknowledged in the dermatologic literature, but should be considered in the differential diagnosis for mucous cysts. PMID- 9857257 TI - [Treatment of isolated testosterone deficiency]. PMID- 9857258 TI - [History of the University Clinic of Dermatology and Venereology in Graz. A review of 125 years (1873-1998)]. PMID- 9857259 TI - [Narrow band UVB 311 nm versus broad band UVB. New developments in phototherapy]. PMID- 9857260 TI - Physiological and clinical significance of carotenoids. PMID- 9857261 TI - The role of carotenoids and retinoids in gap junctional communication. AB - Induction of gap junctional communication (GJC) is discussed as one possible mechanism underlying the cancer-preventive properties of carotenoids. Structurally different carotenoids have different effects on this GJC pathway. Beta-carotene, echinenone, canthaxanthin, cryptoxanthin and 4-hydroxy-beta carotene efficiently induce GJC in murine fibroblasts. Decomposition products of canthaxanthin (e.g. 4-oxo-retinoic acid) enhance both GJC and expression of connexin 43 mRNA. Thus, the biological activity of canthaxanthin is due at least in part to formation of active decomposition products such as 4-oxo-retinoic acid. A number of other small molecules such as 1,25-dihydroxycholecalciferol or thyroid hormones serve as nuclear receptor ligands. Cholecalciferol, 3,3',5 triiodo-L-thyronine and L-thyroxine induce GJC to a similar extent as carotenoids or retinoic acid. Interactions between these signalling pathways may be involved in GJC regulation. PMID- 9857262 TI - Effect of lutein on beta-carotene absorption and cleavage. AB - Carotenoid interactions during absorption and in postabsorptive metabolism have been observed in both human and animal studies. We reviewed the mutual interactions reported between lutein and beta-carotene and report new data on the postprandial beta-carotene and retinyl ester response in the triglyceride-rich plasma lipoprotein (TRL) fraction in volunteers after a single test meal with beta-carotene alone, or beta-carotene combined with lutein or lycopene. Results indicate an inhibitory effect of lutein on beta-carotene absorption, but apparently not on beta-carotene cleavage. In a comparative study with two beta carotene/lutein ratios (2:1 and 1:2, respectively), this inhibitory effect of lutein was found to be most marked when lutein was the predominant carotenoid. In studies on plasma (serum) response also an inhibitory effect of beta-carotene on lutein response was observed. PMID- 9857263 TI - Potential of vegetable processing to increase the delivery of carotenoids to man. AB - It has been suggested that carotenoids contribute to the inverse association between vegetable consumption and risk for coronary heart disease and cancer. However, there are indications that the bioavailability of beta-carotene from vegetables is less than previously thought. Therefore, it is of interest to explore whether the bioavailability of carotenoids from vegetables can be improved. We have reviewed data on the effect of vegetable processing on carotenoid bioavailability. Based on this analysis, we conclude that processing of vegetables by mechanical homogenization or heat treatment has the potential of increasing the bioavailability of carotenoids. Addition of fat during processing may also have an effect. PMID- 9857264 TI - Quantification of the "SLAMENGHI" factors for carotenoid bioavailability and bioconversion. AB - The bioavailability of carotenoids is the fraction of ingested carotenoids utilized for normal physiological functions or storage, while bioconversion is the proportion of bioavailable carotenoid converted to retinol (vitamin A). Factors influencing carotenoid bioavailability and bioconversion have been incorporated in the mnemonic "SLAMENGHI": species of carotenoid; molecular linkage; amount of carotenoids consumed in a meal; matrix in which the carotenoid is incorporated; effectors of absorption and bioconversion; nutrient status of the host; genetic factors; host-related factors; and mathematical interactions. In this paper, an attempt is made to quantify the impact of the various SLAMENGHI factors on carotenoid bioavailability and bioconversion although many of the data available are not strictly comparable. PMID- 9857265 TI - The contribution of various foods to intake of vitamin A and carotenoids in The Netherlands. AB - This study presents data on dietary intake of specific carotenoids in the Netherlands, based on a recently developed food composition database for carotenoids. Regularly eaten vegetables, the main dietary source of carotenoids, were sampled comprehensively and analysed with modern analytic methods. The database was complemented with data from recent literature and information from food manufacturers. In addition, data on intake of vitamin A are presented, which are based on the most recent update of the Dutch Food Composition Table. Intake of vitamin A was calculated for adult participants of the second Dutch National Food Consumption Survey in 1992, whereas intake of carotenoids was calculated for participants of the Dutch Cohort Study on diet and cancer, aged 55 to 69 in 1986. Mean intake of vitamin A amounted to 1.1 and 0.9 mg RE/day for men and women, respectively; the contributions of meat, fats and oils, vegetables and dairy products to total intake were 35%, 24%, 16%, and 16%, respectively. Mean intake of alpha-carotene, beta-carotene and lutein plus zeaxanthin was 0.7, 3.0, and 2.5 mg/day respectively for both men and women, while mean intake of lycopene was 1.0 mg/day for men and 1.3 mg/day for women. The most important foods contributing to intake of beta-carotene and lutein plus zeaxanthin were carrots (beta-carotene only), spinach, endive and kale. PMID- 9857266 TI - Malaria is associated with reduced serum retinol levels in rural Zambian children. AB - Vitamin A deficiency increases the risk of illness, while infections impair vitamin A status. Malaria is highly prevalent in rural Zambia. We describe the relationship between malaria and vitamin A status. We examined dietary vitamin A intake, malaria parasitaemia and serum concentrations of retinol, C-reactive protein (CRP) and alpha 1-acid glycoprotein (AGP) in 210 children under the age of 2. Vitamin A intake was low. Serum retinol was negatively correlated with malaria parasite count and to serum levels of CRP and AGP. Increased malaria parasite density resulted in raised CRP and AGP levels, which were negatively associated with serum retinol. We conclude that improvement of dietary vitamin A intake and prevention of infectious diseases, especially malaria, could alleviate vitamin A deficiency in this population. PMID- 9857267 TI - Impact of a social marketing campaign promoting dark-green leafy vegetables and eggs in central Java, Indonesia. AB - In order to work towards further reduction of vitamin A deficiency in central Java, Indonesia, a social marketing campaign promoting eggs and dark-green leafy vegetables was initiated in March 1996. The nutritional surveillance system (December 1995-December 1996) found the following. The campaign's messages were well noticed. Consumption of at least one egg in the past week increased from 80% to 92% in mothers and from 78% to 92% in children 12-36 months old. It increased in all socio-economic groups and was independent of ownership of chickens. Most eggs had been purchased. The quantity of vegetables prepared increased from 93 to 111 g/person daily and most was purchased. Vitamin A intake increased from 335 to 371 RE/d for mothers and from 130 to 160 RE/d for children. Serum retinol levels increased after the start of the campaign, and were related to egg consumption and vitamin A intake. Because 1. data were collected in such a way that respondents were not aware of the link between data collected and the campaign, and 2. vitamin A status increased and was related to increased consumption of eggs and vitamin A intake, we conclude that the social marketing campaign was successful. PMID- 9857268 TI - Antioxidant effects of carotenoids. AB - Surprisingly, neither the precise pharmacological effect nor the toxicological profile is usually established for food components. Carotenoids are no exception in this regard. Only limited insight into the pharmacology and toxicology of carotenoids exists. It is known that the antioxidant action of carotenoids is determined by 1. electron transfer reactions and the stability of the antioxidant free radical, 2. the interplay with other antioxidants and 3. the reaction with active oxygen. Numerous metabolites of carotenoids are formed upon their action as an antioxidant. Most of these metabolites have an unknown biological activity. It is concluded that a severe lack of knowledge hampers adequate suggestions for human supplementation. PMID- 9857269 TI - Metabolism of retinoic acid: implications for development and cancer. AB - Vitamin A derived retinoids, including all-trans retinoic acid (RA), play an important role in regulating cellular growth and differentiation. Biological activities of retinoids are mediated through interactions with two classes of nuclear receptors, RA receptors (RARs) and retinoid X receptors (RXRs). In addition, cellular retinoid-binding proteins (CRBPs and CRABPs) and RA metabolizing enzymes may explain the pleiotropic effects of retinoids. Recently, a novel cytochrome P450 enzyme (CYP26) with specific RA 4-hydroxylase activity, which is rapidly induced by RA, has been cloned from man, mouse and zebra fish, fullfilling all requirements of an enzyme which could be of crucial importance in controlling steady-state levels of active retinoids in cells and target tissues, thus protecting against excessive exposure. Besides the putative role of this newly identified CYP26 in contributing to susceptibility of cancer cells to retinoids, the possible function of this gene in early embryonic development is discussed. PMID- 9857270 TI - Safety of vitamin A: recent results. AB - A still unresolved public health concern is that excessive vitamin A intake, like vitamin A deficiency, possibly causes birth defects not only in animals but also in man. Due to the low incidence of possibly vitamin A-related malformations in man, available data cannot convincingly define the upper safe limit of periconceptional vitamin A intake. Direct human intervention studies are not feasible for ethical reasons. Therefore, a novel approach in addressing this issue was chosen by combining teratogenicity data from a validated animal model with data on systemic exposure to vitamin A and its major metabolites in female volunteers. In a study in pregnant women endogenous plasma concentrations of vitamin A metabolites during early pregnancy ranged from 0.26 to 7.72 ng/ml. Since they did not cause any foetal malformations, retinoid plasma levels in this range can be considered non-teratogenic. Results of a trial in non-pregnant women document that daily oral vitamin A supplements of 4000, 10,000 and 30,000 IU given for 3 weeks were in the range or slightly above the range of endogenous plasma levels seen in early pregnancy. Even after a 3-week treatment with 30,000 IU/day, peak plasma levels of retinoic acid and isotretinoin were within or just slightly above the range of their physiological levels. In cynomolgus monkeys (average weight: 3-4 kg), a NOAEL (no observed adverse effect level) of 7500 IU per kg body weight and a LOAEL (lowest observed adverse effect level) for developmental toxicity of 20,000 IU/kg was found. Considering these results in the cynomolgus monkey, a dose of 30,000 IU/day should also be considered as non teratogenic in man. PMID- 9857271 TI - Importance and timing of sexual counseling after myocardial infarction. AB - BACKGROUND: This study examined differences in the perceptions of the importance and timing of sexual counseling in patients with myocardial infarction (MI). METHODS: Using a longitudinal descriptive prospective design, subjects were identified from the discharge records of three health-care agencies after having experienced an acute MI in the last 4 to 8 weeks. The Sexual Counseling Needs of MI Patients Survey and a consent form were mailed to patients at 2, 4, and 6 months after MI. Patients (N = 91) rated the importance and timing of 14 sexual counseling items using a Likert scale (1 to 5), and reported the preferred method of education. The possible range for the importance score was 14 to 70. RESULTS: Mean importance ratings for each of the 14 items at each of the data collection periods ranged from 3.28 to 4.59. The mean overall importance score at 2 months (56.96), 4 months (56.52), and 6 months (55.19) reflected the importance of these items for teaching. There was no significant difference between importance scores at each of these time periods. Subjects were divided as to when sexual counseling should occur, in hospital or after discharge, for items related to specific sexual counseling. The most preferred educational method at 2, 4, and 6 months after MI were written materials, followed by individual discussion, and a videotape to watch at home. CONCLUSIONS: Patients validated the importance of each of the specific areas for sexual counseling after MI. The timing and educational methods for sexual counseling must be further addressed in future research. Sexual counseling initiated in the acute-care setting and continued throughout the recovery period can impact the quality of life of patients with MI and their partners. PMID- 9857272 TI - Comprehensive cardiac rehabilitation improves the control of dyslipidemia in secondary prevention. AB - BACKGROUND: Secondary prevention is an important goal of cardiac rehabilitation in patients with coronary heart disease (CHD). Dyslipidemia is one of the major risk factors that is important to control to reduce the incidence of future ischemic coronary events. The aim of the present study was to assess whether control of dyslipidemia, in secondary prevention, could be improved by a comprehensive cardiac rehabilitation program. METHODS: Fifty-two newly diagnosed hyperlipidemic men, who had experienced a recent CHD event, were separated in two equal groups of 26 patients: group CR+, in which patients were included in a 2 month cardiac rehabilitation program including an extensive educational program on cardiovascular risk factors, lipids, and diet, and group CR-without any cardiac rehabilitation. Mean age, body mass index, initial levels of total cholesterol, triglycerides, high-density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol, and LDL/HDL ratio were not significantly different between the 2 groups. In both groups, each patient was referred to a dietitian and to the same lipidologist to start an appropriate hypolipidemic treatment. Treatment of coronary event, type of hyperlipidemia, and hypolipidemic treatment were not different between the two groups. RESULTS: Lipid measurements, performed 3 months after the beginning of the hypolipidemic treatment, showed that patients from the CR+ group, compared with those from the CR-group, had a significantly greater reduction of total cholesterol (23% versus 13%; P < 0.001), of LDL cholesterol (28% versus 12%; P < 0.001), of LDL/HDL ratio (34% versus 13%; P < 0.01) and of triglycerides (33% versus 21%; P = 0.05). CONCLUSIONS: Patients with CHD included in a comprehensive cardiac rehabilitation program showed a significantly better response to the hypolipidemic treatment than patients without cardiac rehabilitation. These results could be attributable to the extensive educational program on secondary prevention performed during cardiac rehabilitation, leading to optimized knowledge on lipid-lowering diet and to improved diet and drug adherence. A secondary prevention educational program must be an important part of any comprehensive cardiac rehabilitation program in patients with CHD. PMID- 9857273 TI - Combined resistive-aerobic training in older patients with coronary artery disease early after myocardial infarction. AB - BACKGROUND AND PURPOSE: Older patients with artery disease have higher rates of disability than younger coronary patients, partly based on lower levels of skeletal muscle strength. We compared the effect of a combined resistive-aerobic training program on muscular strength in older and younger coronary patients early after myocardial infarction. METHODS: The study population included 45 patients who had recently (4-12 weeks) suffered a myocardial infarction; 19 were age > or = 62 years (mean 68 +/- 3 years) and 25 were < 60 years of age (mean 48 +/- 7 years). Muscle strength was measured by single repetition maximum lifts for leg extension and bench press, before and after the 12-week conditioning program. Body composition was measured in a subset of 16 patients by dual x-ray absorptiometry. RESULTS: At baseline, the older population demonstrated lower strength measures than the younger patients. With conditioning, both groups improved strength similarly with leg-extension and bench press measures increasing by 35% and 14% respectively in the older patients and 39% and 14% in the younger patients (both P < 0.05, NS between groups). Within the older patient group, the men were significantly stronger than the women at baseline yet the women tended to improve their strength measures to a greater degree than the men, increasing leg strength by 66% and bench press by 29% versus 29% and 10% in older men (P < 0.10 between groups). In the overall study population, body composition measures showed a slight decrease in body weight, a decrease in fat mass, and a maintenance of lean body mass and bone mineral content with no difference in response between older and younger patients. CONCLUSIONS: Older coronary patients can effectively increase body strength with a combined resistive-aerobic exercise program in the early post-myocardial infarction period. PMID- 9857274 TI - Exercise prescription for patients with pacemakers. AB - When developing an exercise program for pacemaker patients, basic information about the pacemaker must be understood. Atrial, ventricular, and dual-chamber devices can produce varying exercise responses and impact the exercise prescription. The type of rate adaptive sensor the pacemaker has will affect the nature of heart rate response, and therefore, must be taken into account when prescribing exercise. While rate modulation is used with most chronotropically incompetent patients, individuals with VVI pacemakers will also benefit from regular exercise. Although the value of exercise testing pacemaker-dependent patients for ECG interpretation may be limited, it is useful in determining exercise capacity and ensuring proper pacemaker function. Participation in a supervised exercise training program can greatly enhance the follow-up and management of pacemaker-dependent patients as well as afford them the opportunity to experience the physical and psychologic benefits typically associated with cardiac rehabilitation. PMID- 9857275 TI - Chronic obstructive pulmonary disease: capillarity and fiber-type characteristics of skeletal muscle. AB - BACKGROUND: The purpose of this investigation was to compare capillarity and fiber type proportions of the vastus lateralis muscle between patients with chronic obstructive pulmonary disease (COPD) and healthy subjects. METHODS: Fifteen male subjects were included in the study (8 COPD: 61.0 +/- 1.8 years [mean +/- SEM]; forced expiratory volume in 1 second 42.0 +/- 2.1% predicted; 7 N: age 54.0 +/- 1.1). Subjects were submitted to a symptom-limited maximal exercise test on ergocycle. After a transcutaneous biopsy of the vastus lateralis muscle, sections were cut 8 to 10 microns thick and stained with the Andersen method for capillarity and Stevens method for fiber typing. RESULTS: Patients with COPD had a decrease in peak oxygen consumption compared with healthy subjects (1.2 +/- 0.1 versus 3.0 +/- 0.2 L/min). Number of capillaries per square millimeter was lower in patients with COPD versus healthy subjects (92.6 +/- 16.1 and 213.3 +/- 33.5, P < 0.001); percentages of fiber types were 43.5 +/- 5.5% type I, 56.5 +/- 5.5% type II in COPD, and 56.7 +/- 3.4% type I, 43.2 +/- 3.4% type II in healthy subjects (P < 0.05). In addition, capillaries/fiber ratio was 0.83 +/- 0.05 in COPD, and 1.56 +/- 0.10 in healthy subjects (P < 0.001). CONCLUSION: As expected, patients with COPD showed a decrease in exercise capacity. The muscle analysis results indicate that patients with COPD have a greater proportion of type II fibers and a much lower capillaries/fiber ratio than normal subjects. We conclude that COPD adversely affects fiber type and capillarization of the lower limbs. This could be partly caused by deconditioning in these patients. PMID- 9857276 TI - Evaluation of a new standardized ramp protocol: the BSU/Bruce Ramp protocol. AB - BACKGROUND: Because of recent technological advances, exercise testing laboratories now have the ability to use ramp protocols with treadmill exercise tests. Since the Bruce protocol is the most widely used treadmill protocol in clinical laboratories, a standardized ramp treadmill protocol was developed that corresponds to the speed and grade settings of the Bruce protocol at each 3 minute time interval. The purpose of this study was to evaluate the utility of using subject demographic and exercise test data to predict peak oxygen uptake (VO2peak) for tests conducted with the BSU/Bruce Ramp protocol. METHODS: Maximal exercise tests were performed by 698 men and women using the BSU/Bruce Ramp protocol. Peak oxygen uptake was measured during all tests. Stepwise multiple regression analyses were used to predict VO2peak (mL.kg-1.min-1) from maximal treadmill test time and selected variables including age, gender, physical activity habits, and body weight. RESULTS: Maximal test time was found to be the most potent predictor of VO2peak, accounting for 86% of the variance in peak aerobic power, with a standard error of estimate of 3.4 mL kg min-1. A multiple regression equation including age, gender, physical activity habits, and body weight resulted in a slightly improved prediction (R2 = 0.88; standard error of estimates = 3.1 mL kg min-1). CONCLUSIONS: Peak oxygen uptake values can be predicted with reasonable accuracy from the BSU/Bruce Ramp protocol. The BSU/Bruce Ramp would be an excellent choice for laboratories desiring to use a ramp treadmill protocol because of the design of the protocol with identical workloads at equivalent time periods (3, 6, 9, 12, 15, 18, 21 minutes) as the commonly used Bruce protocol. PMID- 9857277 TI - Gender differences in psychosocial profile at entry into cardiac rehabilitation. AB - PURPOSE: To find out in a sample of coronary patients at entry into a cardiac rehabilitation program, whether women report a lower level of physical functioning and a higher degree of psychosocial impairment than men. METHODS: One hundred and nine male and 122 female patients matched for age with a diagnosis of myocardial infarction, coronary artery bypass grafting, or coronary angioplasty were assessed at entry into a multidisciplinary outpatient rehabilitation program. Information was retrieved from hospital records, a semistructured interview, and several psychological questionnaires. Variables of physical functioning included perceived exercise tolerance, functional complaints, psychosomatic complaints, and sleeping disturbances. Variables of psychosocial impairment included anxiety, depression, vital exhaustion, well-being, feelings of being disabled, displeasure, and social inhibition. RESULTS: At program entry, there were no significant differences in age, coronary risk factors, coronary incident, or medication between genders. However, women reported significantly lower perceived exercise tolerance and significantly more functional and psychosomatic complaints. Moreover, women were significantly more anxious and scored significantly higher on social inhibition and vital exhaustion than men. CONCLUSIONS: At entry into rehabilitation, women in this sample report a higher degree of psychosocial impairment and a lower level of physical functioning than men. Women's significantly higher scores on social inhibition and vital exhaustion might partly explain their reported higher drop-out and lower adherence rates in cardiac rehabilitation. PMID- 9857279 TI - Cardiac rehabilitation without exercise tests for post-myocardial infarction and post-bypass surgery patients. AB - PURPOSE: To compare the progress of patients who were exercise tested before or during cardiac rehabilitation versus those patients who were not tested. METHODS: Eighty-eight (88) post-myocardial infarction patients and 141 post-bypass surgery patients had a symptom-limited exercise test before or during 12 weeks of cardiac rehabilitation. Another 125 post-myocardial infarction and 146 post-surgery patients were not tested. RESULTS: Caloric expenditure during class increased for the entire group (P < 0.001) from week 1 to week 12. Body weight decreased for the entire group as a result of cardiac rehabilitation (P < 0.001). Tricep skinfolds decreased for the entire group (P < 0.001) while subscapular skinfolds did not change (P = 0.28). The percent change from week 1 to week 12 for both groups was similar for all variables. No problems occurred during cardiac rehabilitation that required emergency medical management. CONCLUSIONS: Patients completing a 12-week cardiac rehabilitation program can be safely progressed in terms of their exercise capacity without an entry exercise test. This is desirable in a managed-care setting for reducing costs while maintaining effective patient care. Such factors as staff training and experience, institutional philosophy, patient referral patterns, and facility location must be considered before adopting a no-test policy. PMID- 9857278 TI - Influence of intensive physical training on urinary nitrate elimination and plasma endothelin-1 levels in patients with congestive heart failure. AB - BACKGROUND: Congestive heart failure (CHF) is associated with increased peripheral vascular resistance. Exercise-induced shear stress may release endothelial relaxing factors, such as nitric oxide (NO), and inhibit the production of vasoconstrictors such as endothelin-1 (ET-1) thereby modulating vascular tone. We examined the effect of intensive training on ET-1 plasma concentrations and NO-metabolite elimination in patients with CHF after acute myocardial infarction. METHODS: Seventeen patients with CHF after a myocardial infarction were randomized to an exercise group (n = 9), who performed physical training for 8 weeks, or a control group (n = 8) who received usual care. A physical examination, pulmonary function test, and a maximum exercise test were performed, and 24-hour urinary nitrate elimination and ET-1 in plasma were determined before and at the end of the study period. RESULTS: Maximal oxygen uptake remained unchanged in controls (17.9 +/- 1.4 to 18.1 +/- 1.5 mL/(kg min) but increased in the exercise group (from 20.4 +/- 0.75 to 26.7 +/- 1.4 mL/(kg min). After 8 weeks the urinary nitrate elimination in controls was significantly decreased (1.25 +/- 0.20 to 1.03 +/- 0.22 mmol/24 hours; P < 0.001), while it was unchanged in the exercise group (1.26 +/- 0.23 to 1.39 +/- 0.28; P = 0.71). Plasma ET-1 levels did not change after 8 weeks (7.87 +/- 0.62 versus 7.57 +/- 0.75 and 7.13 +/- 0.6 versus 7.35 +/- 0.7 pg/mL for control and exercise groups, respectively). CONCLUSION: In patients with CHF after acute myocardial infarction nitrate elimination decreases over the subsequent 2 months. This trend was reversed by training. Because nitrate elimination mirrors endogenous NO production, these results suggest that training may positively influence endothelial vasodilator function. PMID- 9857280 TI - Exercise testing in cardiac rehabilitation. Is the engine running hot? PMID- 9857281 TI - Probing lymphocyte biology by genomic-scale gene expression analysis. AB - The identity and abundance of mRNA species within a cell dictate, to a large extent, the biological potential of that cell. Although posttranscriptional mechanisms modify protein expression in critical ways, cellular differentiation requires key changes in gene transcription, as evidenced by the potent phenotypes that result from disruption of transcription factor genes in mice. It is now possible to assess the mRNA profile of a cell globally using recently developed genomics techniques. This review focuses on the potential of cDNA microarrays to define gene expression in lymphoid cells, a field which is in its infancy. Examples of cellular activation genes and cytokine inducible genes discovered using this technology are presented but these represent only a taste of the fruit that this new technology will ultimately bear. Gene expression profiles should provide essential new insights into lymphocyte differentiation and activation, the pathogenesis of immune disorders, and the molecular abnormalities in lymphoid malignancies. PMID- 9857282 TI - Antibodies to beta 2-glycoprotein-I: urea resistance, binding specificity, and association with thrombosis. AB - The aim of the present study was to evaluate the urea resistance and binding characteristics of anti-beta 2-glycoprotein I (anti-beta 2GPI) antibodies using standard anticardiolipin (aCL) and anti-beta 2GPI enzyme immunosorbent assays (ELISAs). Sera from patients with antiphospholipid syndrome (APS) (n = 22) and non-APS (n = 24), positive in a standard aCL ELISA, were tested in an anti-beta 2GPI ELISA performed in polystyrene-irradiated ELISA plates. Urea resistance aCL and anti-beta 2GPI ELISAs were performed by measuring the ability of antibodies to recognize antigen in the presence of 2 M urea. The serum dilution after urea treatment (D) expressed as a percentage of the serum dilution without urea treatment (D(o)) corresponding to the same optical density was defined as residual activity (RA = 100 D/D(o)). The higher the RA, the higher the resistance of the antibodies to urea. APS compared to non-APS sera had higher aCL binding (absorbance values ranging between 0.180 and 1.400; median, 0.717 vs 0.120-1.273; median, 0.250, respectively; P < 0.004). Six APS patients' sera had low aCL levels but they expressed RA > or = 30%. Anti-beta 2GPI antibodies were detected in 15 of 22 APS vs 3 of 24 non-APS patients (P < 0.03); RA > or = 30% was detected in 15 of 22 APS vs 1 of 23 non-APS patients (P < 0.004). Using a CL affinity column, antibodies were purified from three APS anti-beta 2GPI negative and three non-APS anti-beta 2GPI-positive patients and tested in a aCL ELISA, using highly purified bovine serum albumin (BSA) as a blocking agent (modified ELISA); reactivity was not detected in two APS and one non-APS sera. On the contrary, the reactivity of the purified antibodies was high when beta 2GPI was incubated with CL in the ELISA plates; thus some anti-beta 2GPI negative sera from APS patients recognized the CL/beta 2GPI complex, rather than CL or beta 2GPI alone. In conclusion, anti-beta 2GPI antibodies are common in the APS patients, but a number of such patients recognize the CL/beta 2GPI complex and not CL or beta 2GPI. Antibodies to either beta 2GPI or the CL/beta 2GPI complex derived from APS sera present a high resistance to urea. Anti-beta 2GPI antibodies of low urea resistance exist in a minority of non-APS patients with autoimmune disease. PMID- 9857283 TI - Expression of CTLA-4 molecule in peripheral blood T lymphocytes from patients with systemic lupus erythematosus. AB - CTLA-4 is a cell surface molecule expressed on activated T cells that is suggested to deliver a negative signal for T cell activation. Since CTLA-4 might be a negative regulator of autoimmune diseases, we investigated its expression on T cells from 20 patients with systemic lupus erythematosus (SLE) by flow cytometric analysis and RT-PCR. We found that although CTLA-4 mRNA was readily detected in all patients and controls, only a very minor subset of T cells expressed detectable surface CTLA-4 molecules in both groups. But patients with SLE had significantly increased percentages of CTLA-4-positive T cells compared with normal controls, implying at least that there was no apparent defective expression of CTLA-4 molecule in human lupus. The kinetics of CTLA-4 expression on T cells stimulated in vitro with PMA plus ionomycin were similar in normal controls and patients with SLE. The expression of CTLA-4 molecules after stimulation increased gradually and peaked at 72 hr. However, the induction of CTLA-4 expression on patients' T cells appeared to be weaker than that of normal individuals. Whether this reflects impaired downregulation by CTLA-4 molecules in SLE patients needs to be clarified further. PMID- 9857284 TI - Increased production of a Th2 cytokine profile by activated whole blood cells from rheumatoid arthritis patients. AB - T cells produce regulatory cytokines which control inflammation. In rheumatoid arthritis (RA), a Th1 cytokine profile has been described in the synovium. In order to assess the Th1/Th2 cytokine balance in blood, a one step culture immunoassay procedure was used to measure the ex vivo production of IFN gamma and IL-4 by whole blood cells from 26 RA patients and 25 controls. For comparison, the same cytokines were measured by ELISA in supernatants of activated whole blood cells. The direct whole blood assay was 10-fold more sensitive than standard ELISA to measure IL-4 levels. IL-4 production was higher in RA patients than in controls, whereas that of IFN gamma was lower. Accordingly, the IL-4/IFN gamma ratio, which reflects the Th2/Th1 cytokine balance in blood, was higher in RA patients (P < 0.0001). The present findings indicate a Th2-over-Th1 cytokine balance profile in RA blood. These results are in contrast with the Th1-over-Th2 pattern previously found in the joint, indicating that the two compartments appear to be differently regulated. PMID- 9857285 TI - Expanded T cell populations in patients with Wegener's granulomatosis: characteristics and correlates with disease activity. AB - Patients with Wegener's granulomatosis have a high prevalence of expanded populations of CD4+ and CD8+ T cells bearing different alpha/beta T cell receptors. To elucidate the role of these populations, we studied the phenotypic and functional characteristics of 13 expanded T cell populations in four patients for a period of 35-51 months. The expanded populations generally showed a persistently high expression of the activation markers HLA-DR and CD25. This expression was independent of the activity of the disease. The expanded populations also expressed CD45RO and/or CD45RA and most of them expressed CD57 but not CD28. Analysis of intracellular presence and secretion of IFN-gamma, IL 2, and IL-4 showed that most of the expanded cell populations contained and/or secreted more of these cytokines than the nonexpanded populations, with an especially high expression/secretion of IFN-gamma and IL-2. The expanded populations showed little proliferative response to Con A and OKT3. The proliferative response of the cells was partly restored after preincubation in medium alone. Some of the expanded populations were associated with disease activity, thus suggesting a link between expanded T cells and the disease. The activated status of the expanded populations and the tendency for certain populations to correlate in magnitude with disease activity suggest their involvement in the disease process. The relative stability of these cell populations indicates that the stimulus driving them is persistent, in agreement with the chronicity of the disease. PMID- 9857286 TI - Minimally invasive surgery induces endotoxin-tolerance in the absence of detectable endotoxemia. AB - Lipopolysaccharide (LPS) tolerance is characterized by an impaired proinflammatory cytokine production upon restimulation of mononuclear cells with LPS. LPS is considered the primary activator for this phenomenon. In response to major injury and extensive abdominal surgery, an immune reaction comparable to LPS tolerance has been described. Therefore, it was investigated whether primary stimuli other than LPS could induce cytokine downregulation. In eight patients who underwent a laparoscopic cholecystectomy, blood was obtained before and after induction of anaesthesia and 2, 6, and 24 hr postoperatively. Ex vivo stimulation of whole blood resulted in a transient reduction (nadir 2 hr postoperatively) of tumor necrosis factor-alpha, interleukin (IL)-1 beta, and interferon-gamma release, while IL-1 receptor antagonist production increased. Stress hormones, LPS-binding protein, and bactericidal/permeability-increasing protein do not seem to be involved. This study shows that minimally invasive surgery, in the absence of endotoxemia, can induce LPS desensitization. These data suggest that prior endotoxemia is not essential for the development of LPS tolerance. PMID- 9857288 TI - Leukocyte transfusion-associated granulocyte responses in a patient with X-linked hyper-IgM syndrome. AB - X-linked hyper-IgM syndrome (XHIM) is a severe congenital immunodeficiency caused by mutations in CD154 (CD40 ligand, gp39), the T cell ligand for CD40 on B cells. Chronic or cyclic neutropenia is a frequent complicating feature that heightens susceptibility to severe infections. We describe a patient with a variant of XHIM who produced elevated levels of serum IgA as well as IgM and suffered from chronic severe neutropenia. Eight of ten leukocyte transfusions with cells from a maternal aunt, performed because of mucosal infections, resulted in similar episodes of endogenous granulocyte production. Transfection studies with the mutant CD154 protein indicate that the protein is expressed at the cell surface and forms an aberrant trimer that does not interact with CD40. The data suggest that allogeneic cells from the patient's aunt, probably activated T cells bearing functional CD154, may interact with CD40+ recipient cells to produce maturation of myeloid precursors in the bone marrow. PMID- 9857287 TI - Presence of activated B-1 cells in chronic inflamed gingival tissue. AB - B-1 cells are physically and functionally unique B cells that produce polyreactive natural antibody. This study examined the activation of B-1 cells in inflamed gingival tissue. Serum IgG antibodies to phosphorylcholine, E. coli LPS, DNA, and some commensal bacteria were examined in adult periodontitis patients and healthy subjects. In addition, the proportion of B-1a (CD20+CD5+) cells and the amount of IL-6 and IL-10 in the inflamed gingival tissues were examined. The serum levels of IgG antibodies to phosphorylcholine, E. coli LPS, and commensal bacteria were significantly higher in the adult periodontitis patients than the healthy subjects. The proportion of B-1a cells and the amount of IL-6 and IL-10 were significantly higher in the inflamed gingival tissues than in peripheral blood from the healthy subjects. These results suggest the activation of B-1 cells in the inflamed gingival tissue of adult periodontitis patients, and that B 1 cells may serve as the first line of defense by producing polyreactive antibodies to phosphorylcholine, LPS, and commensal bacteria. PMID- 9857289 TI - TSP, PM10 and PM10/TSP ratios in the Mexico City Metropolitan area: a temporal and spatial approach. AB - Data for PM10 and TSP collected from SEDUE's (Secretary of Urban Development and Ecology) five principal air monitoring stations from March of 1988 to March of 1989 were analyzed with an emphasis on spatial and temporal distributions. The Mexico City sites consisted of a mixed residential and industrial setting in the NW, a highly industrialized area in the NE, the commercial and administrative district downtown, and areas in the SW and SE dominated by residential neighborhoods. High volume samplers for TSP and PM10 running in parallel were used following a protocol similar to the one specified by the U.S. Environmental Protection Agency. The individual data collected ranged from 24 to 1494 micrograms/m-3 for TSP and from 17 to 607 micrograms/m-3 for PM10. The PM10-to TSP ratios ranged from 0.104 to 0.914 for a one year period, 0.495 being the overall mean ratio. Seasonal and geographical differences in this ratio may reflect the range of sources, transport and transformations of primary and secondary particles. High concentrations of particles were found during the winter and early spring, followed by relatively low concentrations during the summer. Statistically significant differences were found for the two most contrasting sites, the NE and SW areas of the city. The northern and eastern parts were the most impacted by particulate matter, while the area at the southwestern edge of the city was least polluted by fine and total suspended particles. Two interesting observations have emerged from this analysis. First, there is the exceedingly high daily and annual values for both TSP and PM10, relative to the Mexican and U.S. standards and WHO criteria for particulate matter. Second, although the SW area of Mexico City had the lowest fine suspended particle concentrations, it is in this area that the highest concentrations of ozone have been observed. PMID- 9857290 TI - Exposure to air pollution from transitional household fuels in a South African population. AB - Exposure to high levels of particulate matter is one of the risks associated with a household utilizing low quality coal as primary household energy source. In South Africa more than 20 million people rely on traditional (wood) and transitional (coal and paraffin) fuels to fulfil their basic energy needs- cooking and space heating. Personal exposures to TSP in a coal burning Township in the Vaal Triangle (central South Africa) were determined during the winter and summer of 1991 and 1992. Personal monitoring of TSP were conducted over a 12 hour period. Forty five children, age 8-12, participated. Exposures varied from 294 to 2304 micrograms/m3. The average 12 hour exposure during a summer day (8:30 a.m. 8:30 p.m.) was 662 micrograms/m3 (N = 15) compared to 1333 micrograms/m3 for a winter school day (N = 13). The difference was highly significant (P < 0.0002). The study identified the following risk factors associated with exposure: season (winter higher than summer), gender (boys higher than girls), and day of the week (school days higher: schools are situated in the centre of completely unelectrified areas). Statistical significant difference between personal exposures to TSP were documented between electrified vs unelectrified (utilize coal only) areas. The study confirmed other reports of extremely high exposures to air pollution documented in developing communities across the globe. PMID- 9857291 TI - A survey of street sellers' exposure to carbon monoxide in Mexico City. AB - The aim of this study was to explore the CO exposure experienced by street sellers working on busy roads in Mexico City. In January 1991, CO measurements were taken at street level of selected avenues in the city centre. A survey of the numbers of street sellers working on the monitored streets, and their working schedules, was also undertaken. Pavement CO levels were compared with the concurrent concentrations measured at the nearest fixed-site monitoring station. Short term pavement CO concentrations ranged from 2.0 to 70.0 ppm with a mean concentration of 26 ppm. There was a significant positive correlation between pavement and fixed-site CO concentrations. CO concentrations at the pavements were consistently higher than the concurrent fixed-site monitor levels; the average ratio of street/fixed-site concentrations was 2.2. There were more than 1000 street sellers working in the surveyed avenues. More than 80% of the sellers reported that they work at least 6 days a week, with an average working shift of 10 hours per day. The findings of this study suggest that street sellers in Mexico City are exposed to CO concentrations well above national and international air quality criteria. Further research is needed to evaluate the health effects caused by CO exposures among such occupationally exposed groups. PMID- 9857292 TI - Real-time measurement of sub-PPM concentrations of airborne chemicals in semiconductor manufacturing. AB - Real-time mass spectroscopy (ICAMS) can provide hourly or daily estimates of employee exposure. Field calibration of the unit indicated essentially linear response from 0.01 (Cellosolve Acetate) and 0.03 ppm (Diglyme) to 1 ppm in semiconductor cleanrooms. The instrument can be programmed for 4 minute readings on a single compound, or for rotation among several chemicals, each requiring 4 minute dwell times for analysis. In contrast to full shift personal sampling methods to measure exposure, ICAMS offers insights into the occurrence of peak exposures. In addition, in the occupational environment ICAMS results can be integrated to estimate full-shift within a zone exposures. Thus, the ICAMS extends measurement sensitivities below those currently available and offers a viable alternative to personal sampling in the semiconductor industry. PMID- 9857293 TI - Human immunotoxicologic markers of chemical exposures: preliminary validation studies. AB - The circulating cells of the immune system are sensitive to environmental contaminants, and effects are often manifested as changes in the cell surface differentiation antigens of affected populations of cells, particularly lymphocytes. In this investigation, we explore the likelihood that variation in the expression of the surface markers of immune cells can be used as an index of exposure to toxic chemicals. We recruited 38 healthy New Jersey men to study pesticides effects: 19 orchard farmers (high exposure); 13 berry farmers (low exposure); and 6 hardware store owners (no exposure). Immunophenotyping was performed assaying the following cell surface antigens: CD2, CD4, CD8, CD14, CD20, CD26, CD29, CD45R, CD56, and PMN. Data were analyzed using univariate and multivariate methods. There were no significant differences among the groups with respect to routine medical histories, physical examinations, or routine laboratory parameters. No striking differences between groups were seen in univariate tests. Multivariate tests suggested some differences among groups and limited ability to correctly classify individuals based on immunophenotyping results. Immunophenotyping represents a fruitful area of research for improved exposure classification. Work is needed both on mechanistic understanding of the patterns observed and on the statistical interpretation of these patterns. PMID- 9857294 TI - Biological monitoring of workers at a recently opened hazardous waste disposal site. AB - A health assessment was performed in a recently opened landfill for toxic waste. The site has received 14,000 tons of hazardous waste, confined in drums or deposited as bulk material, which were left outdoors for seven months. The analysis in some samples showed that the waste is rich in heavy metals, however we did not find high levels of contaminates in air or surface soil in different areas on-site. When compared to a control group, high-risk workers show higher levels of arsenic in urine and hair, and non-specific symptoms (irritability and insomnia). But among groups, we did not find statistical differences in urinary mercury, blood lead, phenol in urine, cadmium in hair or blood, sister chromatid exchange values, lymphocytes proliferation kinetics, liver function tests, and other non-specific symptoms. We considered this project as a background study for human exposure to hazardous waste, providing useful results for the future evaluation of chronic effects in the same population. PMID- 9857295 TI - Risk factors for exposure to arsenic at a hazardous waste site. AB - Communities surrounding the Rocky Mountain Arsenal (RMA), a Superfund site in Colorado, were studied in order to determine whether exposures to arsenic were greater among persons who resided there than among residents of a comparison area. A census was conducted in areas adjacent to the RMA and in a comparison area 12-15 miles distant. From a stratified random sample, 469 persons were interviewed and urine samples obtained. Arsenic was detected in urine from 43 (9.2%) of the 469 persons sampled at a detection limit of 10 ppb. Trace levels of arsenic (detectable, but non-quantifiable) were found in 184 (39.2%) of those persons sampled. Neither the frequency of detection, the arithmetic mean nor the geometric mean values for urine arsenic was found to be statistically different when persons living near the site were compared to persons from the more distant comparison area. Therefore, the data were pooled across the study areas to evaluate risk factors for exposure to arsenic in this population-based sample. Multivariate logistic regression analyses were conducted to evaluate the risk of arsenic exposure associated with variables included in the interviews while controlling for confounding. Pathways for exposure to arsenic were evaluated through analysis of residence history, occupation, hobbies, dietary habits, water supply, housing and activity patterns. Children of Hispanic origin or non-white race, children who drank less than three glasses of water daily, and children who spent more time outdoors had an increased risk of having > or = 10 ppb of arsenic in their urine. Among adults, younger persons, especially those less than 40 years of age, persons of Hispanic origin or nonwhite race, and those employed in occupations where arsenic is likely to be found had an increased risk of having > or = 10 ppb of urine arsenic. Consumption of red wine or fish during the week prior to sampling was associated with trace levels of arsenic in urine. PMID- 9857296 TI - Residential exposure summary methodology for a reproductive health study of multiple hazardous waste sites. AB - Evaluation of associations between residence near hazardous waste sites and rare outcomes requires the consideration of large geographic areas and multiple sources of contamination. For a case-control study of birth defects and solvents, metals and pesticide exposure, the authors rely on previously collected data to rate possible residential exposure from hazardous waste sites. Four pathways of exposure are evaluated: air vapor, air particulates, groundwater ingestion, and groundwater inhalation. Areas within one mile of each site are classified according to 1) the probability of exposure to solvents, metals, and pesticides, and, if available, 2) the relative concentration of contaminants. The probability of exposure (low, medium, or high) for air vapor and particulates depends on the evaporation and soil retention potential of the contaminants, degree of containment, predominant wind direction, and size of the contaminated area. The probability of exposure for groundwater ingestion and inhalation depends on whether there are water supply wells or affected basements within one mile, solubility of the contaminants, direction of groundwater flow, and groundwater sampling results. Relative concentration is based on sampling results for the most commonly sampled media. Exposures are shown on a standard template that defines 25 sectors within a 1 mile radius of each site. The method cannot provide exact estimates of residential exposure, because it relies on data of variable quality. It does provide a basis for evaluating health effects at multiple sites by identifying susceptible residential areas. PMID- 9857297 TI - Chronic respiratory disease associated with long-term ambient concentrations of sulfates and other air pollutants. AB - Seventh-day Adventist (SDA) non-smokers who had resided since 1966 within five miles of their 1977 residence (n = 3914) completed a standardized respiratory symptoms questionnaire in 1977 and again in 1987. For each participant, cumulative ambient concentrations from 1977 to 1987 of suspended sulfates (SO4) in excess of several cutoffs as well as mean concentrations were estimated by interpolating monthly ambient concentration statistics from state air monitoring stations to the individual's residential and workplace zip codes. There were significant associations between ambient concentrations of suspended sulfates and development of new cases of asthma, but not new cases of overall airway obstructive disease (AOD) or chronic bronchitis. Comparison of previous analyses, in this population, of respiratory disease symptoms and total suspended particulates (TSP), ozone, and sulfur dioxide (SO2), and multipollutant analyses of these pollutants with SO4, indicated these results were not due to a surrogate relationship with other air pollutants. Development of definite symptoms of AOD and chronic bronchitis was most strongly related to TSP. PMID- 9857298 TI - The relationship between low-level air pollution exposure and short-term changes in lung function in Dutch children. AB - Following air pollution episodes in the winters of 1985 and 1987, a series of studies was initiated to investigate short-term changes in health status in children in relationship to short-term changes in air pollution concentrations. Air pollution exposure was characterized by measurements of SO2, NO2, PM10, nitrate, sulfate, H+, and HONO. Panels of unselected children were studied in four winters (1987-1991). Lung function was measured repeatedly with spirometry in these children. In the winter of 1990/1991, a panel of children with chronic respiratory symptoms was studied as well, with repeated observations of Peak Expiratory Flow made at the home. The three winters of 87/88, 88/89 and 89/90 were very mild; no air pollution episodes with long range transport from the east occurred. In these three winters, lung function was measured on 86 different days. Altogether, over 800 children participated. H+ concentrations (expressed as H2SO4) were generally below 0.6 microgram/m3, with a maximum of only 3 micrograms/m3. Daily mean SO2 concentrations never exceded 100 micrograms/m3 in this period. Nevertheless, lung function in these unselected children was found to decrease with increasing levels of especially particulate matter air pollution. In the winter of 1990/1991, a minor air pollution episode occurred. Maximum SO2 and PM10 concentrations were 105 micrograms/m3 and 174 micrograms/m3 respectively. This episode was associated with decreased lung function in a group of about 112 unselected children. There was also a relationship with decreased daily PEF readings in a panel of 73 children with chronic respiratory symptoms. PMID- 9857299 TI - Home dampness and molds as determinants of respiratory symptoms and asthma in pre school children. AB - The objective of our study was to assess the occurrence of respiratory symptoms in relation to dampness and molds in dwellings in pre-school children. A population-based cross-sectional study was carried out by mailing a parent administered questionnaire to a random sample of children in a source population consisting of all children aged 1 to 6 years of Espoo, an urban-suburban city (pop. 170,000) in the Helsinki metropolitan area. The study population included a total of 2568 children whose parents filled the questionnaire (response rate 80%). Included were questions on respiratory health, partly modified from ATS-DLD 78-C questionnaire, and potential personal and environmental determinants of the outcome. The determinants of interest were histories of water damage, presence of moisture and visible molds and perceived mold odor at home. The outcomes included persistent cough, phlegm and wheezing, persistent nasal congestion and excretion during the past 12 months and current asthma. The determinant-outcome relations were estimated in the logistic regression controlling for age, gender, parents' education, single parent or guardian, environmental tobacco smoke, gas cooking, pets and type of day care. The occurrence of persistent cough (OR 2.17; 95% CI 1.39, 3.39), phlegm (2.20; 1.27, 3.82), wheezing (2.62; 1.39, 4.39), nasal congestion (1.94; 1.15, 4.98), and nasal excretion (1.43; 0.95, 2.17) during the past year were higher with the presence of any determinant, but the occurrence of asthma (OR 1.10; 0.54, 2.24) was similar compared to the reference group. Mold odor during the past year (ORs from 2.38 to 6.87) and water damage over a year ago (ORs from 2.54 to 8.67) had the strongest association with the respiratory symptoms. A dose-response relation of the occurrence of the symptoms associated with the frequency of days with mold odor was observed. There was also a compatible time sequence between water damage taking place more than a year ago and the occurrence of symptoms during the past year. The results provide further evidence on the importance of home dampness and molds in the etiology of respiratory symptoms and new information on the role of mold odor as a risk indicator. PMID- 9857300 TI - Unintentional carbon monoxide deaths in California from charcoal grills and hibachis. AB - This study was carried out to determine the annual incidence rate of and factors associated with unintentional deaths due to carbon monoxide (CO) poisoning from charcoal grills/hibachis in California during 1979-1988. A search through the California Master Mortality File was conducted to identify all cases with ICD-9 codes related to unintentional CO deaths. Individual coroners' investigation reports were obtained from 42 counties and evaluated by three evaluators. After excluding miscoded and misclassified cases, 444 deaths were judged to be authentic cases of unintentional CO poisoning. Among them, 59 deaths found to be due to the improper use of charcoal grills/hibachis. An increasing but not statistically significant trend was observed over the 10-year period. The highest rates occurred among males, asians, blacks, and middle-aged (20-39) people. Fifty four percent of the deaths occurred in motor vehicles and 46% in residential structures. PMID- 9857301 TI - Mechanisms of the toxic effect of lead. I. Free lead in erythrocyte. AB - In this work, a selection of children was examined based on their free erythrocyte protoporphyrin (FEP) and total blood lead (PbB) contents. Two groups with clear differences in lead sensibility were selected. One group with high FEP levels (deep lead alteration), named "normal," and one group with low FEP levels (discrete lead alteration), named "lead tolerant," and both with similar PbB concentration, were formed. The selection, based on FEP level, showed a correlation with other indicators (neuromotor alterations). A lower activity of the enzymes Glyceraldehyde 3-phosphate dehydrogenase (GA3PD), Glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), and Lactate dehydrogenase (LDH) was found in the normal group when compared to the tolerant group. Therefore, the influence of the lead acetate upon the same erythrocyte enzyme activities was investigated. Lead inhibition of the enzymes GA3PD and G6PD was found, and this inhibition could be related to the free lead content found in the erythrocytes. Inhibition of the enzymes 6PGD and LDH was also found, but these enzymes were not inhibited by exposure to lead acetate. Erythrocyte free lead content was high in the normal group when compared to the lead tolerant group and could be a determinant factor in the biochemical alterations found. Low erythrocyte free lead contents could be an indicator of lead tolerance. TCA precipitation was found to be a useful method to evaluate free lead content; it gave similar results to gel filtration molecular chromathography. PMID- 9857302 TI - Prenatal and postnatal blood lead level and head circumference in children to three years: preliminary results from the Mexico City Prospective Lead Study. AB - Multiple regression modeling was used to explore the relationship between prenatal and postnatal blood lead levels and child head circumference in the first three years of life. Maternal blood lead at 36 weeks of pregnancy and cord blood lead were found to be inversely related to 6 month and 18 month head circumference, respectively. Child 12 month blood lead was inversely related to 36 month head circumference. Limitations of the data set and the statistical techniques were discussed, as were the interpretations of the findings. PMID- 9857303 TI - Paternal exposures and cardiovascular malformations. The Baltimore-Washington Infant Study Group. AB - Possible associations between paternal exposures and cardiovascular malformations were evaluated in the Baltimore-Washington Infant Study, a population based case control investigation of congenital heart disease and environmental factors. Home interviews of case and control parents elicited information on parental home and occupational exposures. Analysis focused on twelve cardiac diagnostic groups and paternal exposures incurred during the six months preceding the pregnancy. Associations were identified between jewelry making and atrial septal defect (Odds ratio: 12.6; 95% confidence interval: 2.3-68.6) and membranous ventricular septal defect (8.1; 2.0-33.3), welding and endocardial cushion defect with Down syndrome (1.8; 1.1-3.0), lead soldering and pulmonary atresia (2.3; 1.1-4.9) and ionizing radiation and endocardial cushion defect without Down syndrome (4.7; 1.7 12.6). Ionizing radiation was found to be associated with endocardial cushion defect with Down syndrome only when father was present at interview (5.6; 1.7 17.9); a similar effect of father at interview was noted for paint stripping in relation to coarctation of the aorta (3.5; 1.5-8.0) and muscular ventricular septal defect (3.5; 1.5-8.5). Also, paint stripping was associated with hypoplastic left heart only in the presence of family history of cardiac defects (11.9; 2.4-60.0). This large study on cardiac diagnostic groups and specific preconceptional exposures provides new leads for further assessment of the role of paternal exposures on adverse pregnancy outcome. PMID- 9857304 TI - Aryl hydrocarbon hydroxylase activity in human placenta and threatened preterm delivery. AB - Induction of aryl hydrocarbon hydroxylase (AHH) activity in the placenta has been well documented. This enzyme may be induced by a variety of Polycyclic Aromatic Hydrocarbons (PAHs) and the AHH inducibility is associated with harmful effects of environmental chemicals. Toxic effects of PAHs in tissues such as placenta have been demonstrated to be due to their metabolites, epoxides, which interact with DNA. Thus, environmental PAHs may be related to its alterations in fetal development. Founded on these findings the PAH metabolites could interfere with the normal course of the pregnancy and may be an aborticide, a teratogen or a carcinogen. We hypothesize that low increased activity of placental Aryl Hydrocarbon Hydroxylase (AHH) may be an important determinant of human fetotoxicity. The present investigation was designed to examine the possible implications of PAH exposure at environmental exposure levels on the normal course of the pregnancy using AHH induction as an indicator of PAH exposure. Threatened Preterm Delivery (TPD) was used as an index of problems in the normal course of pregnancy. A group of forty pregnancies at term with TPD was compared with eighty controls for placental AHH induction. Macroscopic placental examination was also performed. A significant increase in prevalence of placental AHH induction with TPD was shown (Odds-Ratio = 2.8; 95% confidence bounds [1.3 6.2]; chi 2 = 6.7 p < 0.01). No such increases were found associated with placental pathology. When taking into account the group of placenta without basal plate calcifications, the significant increase in prevalence of placental AHH induction with TPD above mentioned was greatly increased (Odds-Ratio = 8.9; 95% confidence bounds [2.4-32.9]; chi 2 = 11.1 p < 0.001) controlling for gestational age. The increase in prevalence of placental AHH induction with TPD disappeared when taking into account the subgroup with basal plate or parenchyma calcifications. It is hypothesized that the high estrogen and progesterone at term may explain these associations. PMID- 9857305 TI - Relationship between low lead exposure and somatic growth in adolescents. AB - The possible effects of moderate lead exposure on human growth and development are not well defined. This study was carried out in order to evaluate the relationship between somatic growth and lead exposure. 418 subjects (202 males and 216 females) aged 11-13 years in good health and living in an area of the northern region of Italy were submitted to the following determinations: blood lead levels (PbB), serum concentrations of gonadotropins (LH, FSH) and sex steroid hormones (DHA-S, T, E2) and anthropometric measurements (stature and weight). The mean blood levels were 85.39 and 70.08 micrograms/1, respectively, for males and females. Significant and negative relationships were found between blood lead levels and stature in 13 years old males and 12 years old females (p = 0.009 and p = 0.006, respectively). Negative relationships between lead in blood and LH and FSH were found only in males with lead levels higher than 90 micrograms/1. Our findings seem to suggest that even for low lead exposure this metal may affect stature growth and gonadotropins levels. PMID- 9857306 TI - Reporting bias related to an environmental hazard. AB - During spring 1984, second and fifth grade schoolchildren living in three Haifa Bay areas on the eastern Mediterranean coast, with different levels of air pollution, were studied. The parents of these children filled out ATS-NHLI (American Thoracic Society and the National Heart and Lung Institute) health questionnaires and the children performed PFT (Pulmonary Function Tests). A trend of higher prevalence of most reported respiratory symptoms was found for schoolchildren growing up in the medium and high polluted areas as compared with the low pollution area. Logistic models fitted for the respiratory conditions that differed significantly among the three residential areas also included background variables that could be responsible for these differences. Relative risks for respiratory conditions calculated from these models were in the range of 1.38 and 1.81 for children from the polluted area as compared to 1.00 for the low polluted area. All the measured values of PFT were within the normal range, with no consistent reduction in PFT for any residential area. During spring 1989, seventh graders (second graders in 1984) were reexamined and a new cohort of fifth grade children was studied, using the same techniques as in 1984. A very significant rise in the prevalence of most reported respiratory symptoms and diseases was observed among both fifth and seventh grade schoolchildren in 1989 compared to 1984, especially in the low and medium polluted areas and less in the polluted area. Changes over time in PFT in the older cohort were similar in the three areas. PFT of fifth graders in 1984 and in 1989 were very similar. The most significant factor in logistic models fitted for the prevalence of respiratory conditions among the studied schoolchildren in 1989, was the subjective attitude of their parents towards the deleterious effects of air pollution on their children's health, and the subjective estimate of their children's exposure to pollution rather than measured exposure. A huge campaign carried out during the survey against the main polluters in the Haifa Bay area caused both public concern and apparently reporting bias. PMID- 9857307 TI - Amoebological study of the atmosphere of San Luis Potosi, SLP, Mexico. AB - A one year round survey was conducted for isolation of free living amoebae (FLA) in the city of San Luis Potosi, (SLP), Mexico, which is placed in a desert environment. Samples were taken by modified impinger method and cultivated in laboratory conditions for FLA isolation following a week period of rehidration. 57 strains were isolated, 39% belonged to Acanthamoeba genus (which is important because it bears opportunistic pathogens that produce amoebic keratitis and granulomatous amoebic encephalitis in humans), 16% to Hartmannella, 9% to Vahlkampfia and the other proportion was divided among 6 other genera. The isolations were more abundant during dry season and the main genera were present in all four stands. The difference among them was the species variety which is discused as connected with abundance of organic wastes and lack of urbanization near the stations. PMID- 9857308 TI - Introduction to misconduct in science and scientific duties. PMID- 9857309 TI - The United States Government scientific misconduct regulations and the handling of issues related to research integrity. PMID- 9857310 TI - Scientific dishonesty: a Danish proposal for evaluation and prevention. AB - The term scientific dishonesty covers the wide scope from consciously fraudulent actions to gross negligence and refers both to character trait and to actions. Scientific dishonesty may occur in any country, but reactions in Europe have been slow to emerge. On the basis of a thorough review, the Danish Medical Research Council therefore has made recommendations on how to handle and how to prevent scientific dishonesty. Suspected cases must be referred directly to a national committee chaired by a High Court judge. An ad hoc investigatory committee may be appointed in each case. If dishonesty is established, the institution then is asked to implement appropriate sanctions and to report on the actions taken to the national committee. Prevention of scientific dishonesty forms an essential part of good scientific practice and must have an impact on research training, requirements for data storage, guidelines for scientific publication, and criteria for evaluating grant proposals and promotions. PMID- 9857311 TI - Integrity in science: administrative, civil and criminal law in the U.S.A. PMID- 9857312 TI - Integrity in science: misconduct investigations in a U.S. university. PMID- 9857313 TI - Questions from the delegates and answers by the panelists concerning "ethics and law in environmental epidemiology". PMID- 9857314 TI - Membrane properties that shape the auditory code in three nuclei of the central nervous system. AB - OBJECTIVE: We investigated if auditory neurons have an intrinsic ability to radically transform auditory signals. METHOD: We surveyed membrane properties that control coding by neurons, identified with intracellular staining or infrared-DIC videomicroscopy, in three stations of the auditory pathway. We used intracellular and patch-clamp techniques in slices, to study the voltage responses to current pulse injections and distinguished voltage-gated conductances with selective blockers. RESULTS: First order spherical bushy cells in the anteroventral cochlear nucleus responded at a short, stable latency with single spikes, due to a perithreshold interaction of Na+ and Ca2+ conductances. Two K+ conductances suppressed firing after this onset-spike. Second-order principal neurons of the lateral superior olive use unspecified mechanisms to secure stable onset latencies but maintained a very regular tonic firing, resulting in a chopper pattern. Other intrinsic properties induced a marked accommodation in spike rate. When depolarized as during alert states, neurons in the medial geniculate body (MGB) of the thalamus fired with variable latencies in a tonic mode. At negative resting potentials characteristic of sleep states, they responded at the onset of a depolarization and the offset of a hyperpolarization with phasic bursts due to a transient low threshold Ca2+ current. In the phasic, but not tonic mode, MGB neurons produced high-threshold Ca2+ spikes that may couple signal transmission to the neuron's metabolism. The three neuron types exhibit analogue computing abilities that transform the same input into entirely different output patterns. Isoflurane anaesthesia induces a current shunt in MGB neurons, radically changing the properties and preventing normal responses. Thus, thalamocortical auditory codes are compromised under anaesthesia. CONCLUSION: At all investigated stations of the auditory pathway, input signals are transformed by activation of voltage-controlled conductances and other intrinsic membrane properties. PMID- 9857315 TI - Fat-plug and paper-patch myringoplasty in rats. AB - OBJECTIVE: The purpose of this study is to investigate the recovery rate in paper patch and fat-plug myringoplasty in rats. METHOD: Small and large perforations were inflicted on the tympanic membranes of 60 rats. Fat-plug and paper-patch myringoplasties were performed to different groups and another group was left for spontaneous healing. RESULTS: We found that for small perforations, the recovery rate was 94.7% in fat-plug myringoplasty, 94.4% in paper-patch myringoplasty, and 66.6% in control group. The recovery rates in large perforations were 52.9%, 56.2%, and 26.6%, respectively. On the other hand, in larger perforations, paper patch or fat-plug myringoplasty have not been found effective. CONCLUSION: We believe that due to significant operational advantages, fat-plug or paper-patch myringoplasty can be suggested for the reconstruction of small and dry perforations of the tympanic membrane. PMID- 9857316 TI - Basal cell carcinoma of the conchal bowl: interdisciplinary approach to treatment. AB - OBJECTIVE: The auricle is the site of only 6% of cutaneous neoplasms, but represents an area of high risk for recurrence. The purpose of this work is to present a series of patients with basal cell carcinoma (BCC) of the conchal bowl, and a review of the literature concerning the management of such patients, and to propose an algorithm for use in the treatment of such cases. METHOD: Nine patients with BCC of the conchal bowl were treated using an interdisciplinary approach. Each patient was assessed by both otolaryngology and dermatology. RESULTS: None of the patients was found to have clinical evidence of significant spread into the external auditory canal (EAC). All were treated with Mohs' micrographic surgery (MMS). In each case, otolaryngology was available to perform en bloc resection of the temporal bone, should the tumour be found to extend significantly into the EAC such that it could no longer be accessed by MMS. None of the patients were found to have such extension. All cases were reconstructed using full-thickness skin grafts. All patients remain free of recurrence at an average follow-up of 12.2 months. CONCLUSION: Based on our experience and a review of the literature, we propose an algorithm for use in the management of BCCs of the conchal bowl. We suggest that an interdisciplinary approach that combines the efforts of otolaryngology and dermatology can provide optimal care by saving patients needless procedures and loss of cosmetically important tissue. PMID- 9857317 TI - A new flap for reliable nasal reconstruction. AB - OBJECTIVE: In this article, we describe a new flap for use in lower nasal reconstruction. METHOD: This technique consists of a planned heminasal transposition rotation flap, with both laterally based and glabellar components. Esthetically favourable incisions are made to follow at the junction between the esthetic subunits. RESULTS: Gratifying early results are achievable, resulting in little postoperative edema due to maintenance of laterally based lymphatic and venous outflow. Likewise, pleasing long-term outcomes have been noted in each of the ten patients in whom we have utilized this flap for nasal reconstruction. Complete flap survival has been noted in every patients. CONCLUSIONS: Our experience with the use of this flap technique suggests that it provides consistently rewarding, esthetically pleasing results in patients with cutaneous defects of the lower half of the nose. PMID- 9857318 TI - Adult epiglottitis: the Toronto Hospital experience. AB - OBJECTIVE: This study was conducted to gain insight into the diagnosis, treatment, and outcome measures in adult patients presenting with acute epiglottitis. METHOD: A retrospective clinical study was conducted on fifty-seven consecutive adult patients presenting with acute epiglottitis over a 13-year period to The Toronto Hospital, University of Toronto. Demographic description of patient population, presenting symptoms and signs, investigations performed, treatment provided, and incidence of complications were described. RESULTS: Soft tissue lateral neck x-rays were abnormal in 88.1% of patients tested but had a 12% false-negative rate. Indirect, direct, or flexible laryngoscopy were the most accurate investigations to establish diagnosis and were not associated with any complications. Four of 45 patients (8.9%) had positive blood cultures for Haemophilus influenzae, with the remainder demonstrating no growth, while 22% of patients (8/36) grew possible pathogenic organisms on throat culture. A rapid clinical course (< 12 hr), evidence of tachycardia, or positive pharyngeal or blood cultures were factors that selected for a group of patients requiring formal airway intervention. CONCLUSIONS: Adult epiglottitis is different from its paediatric counterpart in that organism identification is less common, the clinical course is more gradual, there is less seasonal variation, airway compromise is more uncommon, and conservative airway management can be contemplated in the selected patient. Adult epiglottitis can be managed safely with a selective approach to airway management with airway intervention for any patient with signs of critical airway compromise. PMID- 9857319 TI - Gore-tex augmentation grafting in rhinoplasty--is it safe? AB - OBJECTIVE: The utility of the Gore-tex soft-tissue patch in vascular and hernia surgery over the last 20 years led to its application in facial augmentation and rhinoplasty over the last decade. The early success of initial reported series must be balanced against the long-term problems that have faced all implant materials used in the nose in the past. Gore-tex is attractive to the facial plastic surgeon because of its ease in handling and sculpting, and its plentiful supply. This review analyzes the results of Gore-tex augmentation grafting. METHODS AND RESULTS: In a personal series of 30 rhinoplasty cases with minimum 18 month follow-up, there were three cases of infection, two of which required Gore tex removal, and one seroma requiring Gore-tax removal. In two other patients, the Gore-tex was associated with excessive scar tissue. CONCLUSION: Our results led to the development of guidelines for Gore-tex usage in the nose. Since adopting these guidelines, the authors have not yet experienced any complications with Gore-tex. PMID- 9857320 TI - Positron emission tomography in the evaluation of the negative neck in patients with oral cavity cancer. AB - OBJECTIVE: Proper management of the clinically negative neck (N0) in patients with squamous cell carcinoma (SCC) of the oral cavity (OC) is controversial. Detecting cervical metastasis in these patients is important, because cervical lymph node metastasis is associated with an unfavourable prognosis. Conventional radiographic studies, such as computerized tomography (CT) and magnetic resonance imaging (MRI), may augment physical examination. However, there are still limitations of these modalities in detecting the presence of metastatic cervical disease. Positron emission tomography (PET) is a functional imaging modality that has recently been used in patients to detect head and neck neoplasms. We report using PET in the evaluation of the N0 neck in 11 consecutive patients with SCC of the OC who underwent neck dissection. METHOD: The results of PET scans were correlated with the pathologic findings of 19 neck dissections. RESULTS: Four patients (36%) undergoing 7 neck dissections (37%) had pathologic evidence of disease. Positron emission tomography scans were positive in all of these patients and in all pathologically confirmed cervical metastasis. Seven patients (64%) undergoing 12 neck dissections (63%) had no pathologic evidence of cervical metastasis. Positron emission tomography scans were negative for malignancy in all of these seven patients. In the patient with SCC of the OC with a clinically N0 neck, we found PET to have an overall sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of 100%. Computerized tomography demonstrated 40%, 88%, 67%, 70%, 69%, respectively. In all statistical categories except specificity (p = .1), PET demonstrated statistical significance (p < .05) over CT. CONCLUSION: Positron emission tomography appears to be a promising diagnostic aid that may be applied when evaluating the N0 neck for patients with SCC of the OC. PMID- 9857321 TI - Paediatric hearing loss in rural Pakistan. AB - OBJECTIVE: The present study was designed to generate population-based data on the prevalence and causes of hearing loss in rural Pakistani children. A community screening programme was utilized to identify and evaluate the hearing impaired. METHODS: The study was performed in conjunction with the Ministry of Population Welfare in Sialkot District, Punjab Province. School-aged children between the ages of 5 and 15 years were screened and examined for hearing impairment according to World Health Organization (WHO) protocols. Case-control analysis of audiometric, physical examination, and risk factors for hearing loss were performed, followed by chi-square analyses. RESULTS: A total of 607 children comprised the study population, with an overall point prevalence of hearing impairment of 7.9%. Fifty percent of all hearing loss was conductive in nature, amenable to either medical or surgical therapy. The risk factors most associated with conductive hearing loss were otorrhea and multiple ear infections greater than 5. In cases of severe hearing loss, 70% were the result of consanguinous marriages. Almost no cases of hearing loss were attributable to measles, mumps, rubella, and the TORCH infections. CONCLUSION: This study has generated some badly needed population-based data on the magnitude of the problem of hearing loss in rural Pakistan. It is hoped that the results of this work will stress the importance of hearing health in Pakistan and to encourage other professionals to pursue similar projects. PMID- 9857322 TI - Increased otoacoustic-emission amplitude secondary to cochlear lesions. AB - OBJECTIVE: The measurements of transient evoked otoacoustic emissions and distortion-product otoacoustic emissions are being used increasingly, both as an objective hearing test clinically, and as a research tool to investigate the micromechanical aspects of cochlear function. We hypothesized that localized damage in the apical or middle cochlear turns may have an influence on the micromechanics and the function of adjacent, apparently normal cochlea. For that purpose, we used an animal model of localized apical and middle-turn cochlear lesions. METHOD: Extent of damage was assessed by scanning electron microscopy and the function of the damaged cochlea by change in the otoacoustic emission (OAE) levels. RESULTS: We found that localized damage to the apical or middle turn may be accompanied by an increase in OAE measured from adjacent apparently normal cochlea. CONCLUSION: Explanations to this phenomenon are suggested, and possible clinical associations such as to Meniere's disease and to sudden hearing loss are reviewed. PMID- 9857323 TI - Metastatic Sertoli-Leydig cell ovarian cancer manifested as a frontal sinus mass. PMID- 9857324 TI - Retropharyngeal lipoma causing severe dysphagia and dyspnea. PMID- 9857325 TI - Iron supplementation. AB - Iron deficiency affects approx. 20% of the world population. Due to predominantly vegetarian diets that reduce the bioavailability of food iron drastically, deficiency states are most widely distributed in developing countries. In addition, iron demand is increased by blood losses and by fast growth which increases the risk of iron deficiency in infants, young adolescents, and in menstruating and pregnant women. The symptoms of iron deficiency include impaired physical and intellectual performance. Iron supplementation may help to break the vicious cycle between inadequate nutrition and poverty. Fortification programs have to consider social and health aspects, including provision against iron overload. Excess iron stores may promote cancer and increase the cardiovascular risk, though the latter is a subject of current debate. The best approach to control such risks is individual iron supplementation geared to the demand by adequate laboratory controls. However, this approach is too costly for general application in developing countries. Food-iron fortification has successfully reduced iron deficiency in many trials and, in comparison, is much cheaper. As iron deficiency is widely distributed in most developing countries, the risk of inducing iron overload in the general population is low. Genetically determined diseases that may lead to siderosis, such as hereditary haemochromatosis or thalassaemia major, show a limited geographic and ethnic distribution. Such subgroups can be largely avoided by targeting food-iron fortification to infants, young adolescents, or pregnant women. Food vehicle and iron compound have to be matched in order to optimise iron bioavailability and to avoid rancidity in food, spoiling its taste and odour. The fortification of salt, sugar and spice mixtures or of bakery products with a short shelf-life are valid approaches to this end. Alternatively, haem iron can be used to fortify cereal-based food staples in developing countries such as tortillas or chappaties. Thus, a variety of options is available to solve the technical problems of food iron fortification. However, optimal solutions have to be tailored to the individual situation in each country. PMID- 9857326 TI - Failure of IGF-I infusion to promote growth in Zn deficient hypophysectomized rats. AB - The retarded growth of zinc-deficient rats is associated with low plasma insulin like growth factor-I (IGF-I). To determine whether the low circulating IGF-I concentrations are responsible for the stunted growth, the growth response of zinc-deficient hypophysectomized rats to supplementation with recombinant human IGF-I (hIGF-I) was investigated. There were three dietary groups: zinc-deficient (0.9 mg zinc/kg diet), controls (66 mg zinc/kg diet) and zinc-adequate pair-fed (66 mg zinc/kg diet). All rats in each group received hIGF-I (150 micrograms/day) by subcutaneous infusion for 12 days, except for half of the animals in the control group which were sham-treated and which received vehicle infusion for the same period of time. The magnitude of the change was small, nevertheless infusion of hIGF-I significantly increased (p < 0.05) the body weight, tail length, liver weight and tibial epiphyseal width of control animals. This growth effect of hIGF I was inhibited by lack of zinc in the diet, as evidenced by continuous weight loss, no increase in tail length and lowered tibial epiphyseal width in zinc deficient animals. We conclude that systemic supplementation of hIGF-I can promote growth in hypophysectomized rats. The absence of weight gain and bone growth in zinc-deficient animals during the infusion period suggests that the growth promoting effect of circulating IGF-I is impaired by zinc deficiency. PMID- 9857327 TI - Calcium absorption from small soft-boned fish. AB - The prevalence of osteoporosis in developing countries is low compared to most industrialised countries despite an apparent low Ca intake. It is possible, however, that food surveys have overlooked important Ca sources in developing countries. Small fish eaten with the bones can be a rich source of Ca, even though Ca from bone may be considered unavailable for absorption. In the present study, absorption of Ca from indigenous Bengali small fish was compared with the Ca absorption from milk. Ca absorption from single meals was determined in 19 healthy men and women (21-28 y). Each subject received two meal types on two separate occasions. Both meals consisted of white wheat bread, butter and ultra pure water with the main Ca source being either small Bengali fish (397 mg Ca in total) or skimmed milk (377 mg Ca in total). The meals were extrinsically labelled with 47Ca, and whole-body retention was measured on day 8, 12, 15 and 19 after intake of each meal. The labelling procedure was evaluated by an in vitro method. The calculated absorption of Ca as measured with 47Ca whole-body retention was 23.8 +/- 5.6% from the fish meal and 21.8 +/- 6.1% from the milk meal (mean +/- SD), which was not significantly different (p = 0.52). Even after correction for an incomplete isotope exchange, as indicated by the in vitro study, Ca absorption was similar from the two meal types. It was concluded that Ca absorption from small Bengali fish was comparable that from skimmed milk, and that these fish may represent a good source of Ca. PMID- 9857328 TI - Effect of diabetes on the tissular Zn/Cu ratio. AB - One of the parameters related to the development of coronary disease in diabetic patients is the tissular Zn/Cu ratio. We evaluated the levels of Zn and Cu, and the Zn/Cu ratio in insulin target tissues in diabetic and normoglucemic growing Wistar rats in order to determine the influence of diabetes and the disease evolution period. Diabetes was induced chemically by administration of streptozotocin. In order to determine the influence of the duration of diabetes on the Zn/Cu ratio, three time periods were studied: 7, 21 and 60 days. The animals were subsequently sacrificed and the target tissues (liver, adipose tissue, and skeletal muscle) were removed. Zn and Cu levels were measured by AAS after wet mineralization. STZ-induced diabetes modified the tissular Zn and Cu content. There was a significant decrease (p < 0.01) in liver and adipose tissue, but not in skeletal muscle. The in adipose tissue and skeletal muscle, but not in liver, effects were dependent on the duration of diabetes. The Cu content was higher in the liver of diabetic rats (p < 0.1) and lower in adipose tissue (p < 0.1) and skeletal muscle (NS). Tissular Cu levels also were affected significantly by the duration of diabetes. The Zn/Cu ratio showed a generalized decrease, except in skeletal muscle. This decrease was dependent on the presence of diabetes mellitus and the duration of the disease (p < 0.01). PMID- 9857329 TI - Changes in the concentrations of trace elements in human milk during lactation. AB - Inductively coupled plasma mass spectrometry (ICP-MS) was used to determine 18 trace elements (Ba, Be, Bi, Cd, Co, Cs, Cu, La, Li, Mn, Mo, Pb, Rb, Sb, Sn, Sr, Tl, and Zn) in 55 human milk samples from 46 healthy mothers collected during lactation periods extending to 293 days after birth. Se was quantified by hydride generation atomic absorption spectrometry (HG-AAS). To test the accuracy and the precision of the analytical procedure, milk powder reference materials (BCR 063 and BCR 150) were analyzed. The results obtained by ICP-MS and HG-AAS showed good agreement with the certified values. Whenever available, trace element concentrations determined in the human milk samples were compared to reliable literature data. The concentrations of Be (< 0.05 to 0.9 microgram/kg), Bi (< 0.09 to 2.0 micrograms/kg), Cs (1.7 to 7.7 micrograms/kg), La (< 0.05 to 3.7 micrograms/kg), Rb (440 to 1,620 micrograms/kg), and Tl (< 0.08 to 0.5 microgram/kg) are the first to be reported for human milk. The concentrations of the essential trace elements Cu (p < 0.005), Mn (p < 0.05), Mo (p < 0.0005), Se (p < 0.001), and Zn (p < 0.0005) significantly decreased and the concentrations of cobalt significantly increased (p < 0.005) in human milk during the course of lactation. All concentrations for the essential trace element tin in the human milk samples were below the method detection limit of 0.3 microgram/kg. Among the not essential and toxic elements-with the exception of Ba, Pb, and Tl-the trend toward lower concentrations with continuing lactation is much less pronounced than for the essential trace elements. With the exception of Se, the daily intakes of essential trace elements of fully breast-fed infants are considerably lower than dietary recommendations. PMID- 9857330 TI - The TSH-dependent variation of the essential elements iodine, selenium and zinc within human thyroid tissues. AB - Instrumental Neutron Activation Analysis was used in order to measure iodine, selenium and zinc concentration in thyroid samples. A pair of samples of normal and nodular tissue were collected from the thyroid gland from 72 patients selected on the basis of pathological criteria (44 cases of multinodular goiter, 12 of chronic lymphocytic thyroiditis (CLT), 6 of thyroid adenoma (TA) and 12 of thyroid cancer (TC)). The check for tissue homogeneity and sampling error was performed by means of the coefficient of variation (CV%) of the elements in replicate samples of normal and altered tissues. High CV% values (> 15%) for iodine reflected a functional variability in thyroid follicles, while low CV% values (< 10%) for selenium and zinc indicated that the composition of selected tissues was rather homogeneous. The variation of the element's concentration was compared in normal and altered tissues. The mean element concentrations had values close to those already reported in the literature; furthermore, our patients had marginal iodine and selenium deficiency. Both normal and nodular tissues in CLT showed statistically significant lower zinc values as compared with the other thyroid diseases. To evaluate the thyroid function, thyroid stimulating hormone (TSH) and thyroxine (T4) levels were measured in the serum of patients. Two arbitrary serum-TSH threshold levels (TSH < 1.0 and > 4.0 mU/L) were introduced in order to classify, respectively, hyperthyroidism and hypothyroidism, as well as euthyroid conditions (1.0 < TSH < 4.0 mU/L), and each patient was assigned to one of these groups. The influence of TSH in the variation of the concentration of iodine, selenium and zinc in normal and altered human thyroid tissues was significant. PMID- 9857331 TI - Review of publications. PMID- 9857332 TI - Pharmacokinetic aspects of measurement of glomerular filtration rate in the dog: a review. AB - Glomerular filtration rate (GFR) is estimated by means of clearance, defined as the volume of plasma that has been cleared of a particular substance per unit time. Glomerular filtration rate may be estimated by measuring the renal clearance of a filtration marker using data from both urine and plasma or by plasma clearance using only plasma data. Several alternative pharmacokinetic models are used for the calculation of clearance using various filtration markers with slightly different pharmacokinetic properties. The purpose of this article is to discuss how the choice of marker and pharmacokinetic model may influence estimated GFR values and to elucidate commonly used methods and reported GFR values in the dog. PMID- 9857333 TI - Diagnostic features of clinical neurologic feline infectious peritonitis. AB - Feline infectious peritonitis (FIP) is a fatal Arthus-type immune response of cats to infection with FIP virus, a mutant of the ubiquitous feline enteric coronavirus (FECV). The disease may occur systemically or in any single organ system, and primary neurologic disease is a common subset of such manifestations. We examined 16 domestic cats with clinical neurologic FIP and 8 control cats with nonneurologic FIP, with the intention of identifying the ante- and postmortem diagnostic tests that most contribute to accurate diagnosis. Of the 16 cats with neurologic FIP, 15 were less than 2 years of age and all 16 originated from large multiple-cat households. The most useful antemortem indicators of disease were positive anti-coronavirus IgG titer in cerebrospinal fluid, high serum total protein concentration, and findings on magnetic resonance imaging suggesting periventricular contrast enhancement, ventricular dilatation, and hydrocephalus. Postmortem diagnosis was facilitated by FIP monoclonal antibody staining of affected tissue and coronavirus-specific polymerase chain reaction. Most cats with neurologic and ocular forms of FIP had patchy, focal lesions, suggesting that recently developed technologies described in this report may be useful for evaluation of cats with suspected FIP. PMID- 9857334 TI - Nemaline rods in canine myopathies: 4 case reports and literature review. AB - The diagnosis of nemaline rod myopathy (NM) is based on the presence of numerous pathognomonic rods within a fresh frozen muscle biopsy specimen. Three forms of congenital NM have been described in humans, and rods have been found to occur in various other conditions. A similar myopathy was described in 1986 in a family of cats. In this report, we describe a case of congenital NM in a 10-month-old Border Collie, an adult-onset NM in an 11-year-old Schipperke, and 2 acquired myopathies with nemaline rods in adult dogs associated with hypothyroidism and Cushing's syndrome. Common clinical features included exercise intolerance, abnormal electromyography, and the presence of nemaline rods in fresh, frozen, and glutaraldehyde-fixed biopsies from proximal appendicular limb muscles. Staining of cryostat sections of muscle biopsy specimens by the modified Gomori trichrome technique disclosed numerous rod bodies that were localized to type 1 fibers by the histochemical adenosine triphosphatase reaction. Accumulation of rods also was demonstrated by electron microscopy in 2 of the cases with localized enlargement and streaming of Z lines. Documentation of NM in a young Border Collie and the adult-onset form in the Schipperke alerts clinicians to the existence of this disorder in these breeds. PMID- 9857335 TI - Influence of veterinary care on the urinary corticoid:creatinine ratio in dogs. AB - Physical and emotional stresses are known to increase the production and secretion of glucocorticoids by the adrenal cortex in both humans and experimental animals. The urinary corticoid: creatinine (C:C) ratio is increasingly used as a measure of adrenocortical function. In this study we investigated whether a visit to a veterinary practice for vaccination, a visit to a referral clinic for orthopedic examination, or hospitalization in a referral clinic for 1.5 days resulted in increases of the urinary C:C ratio in pet dogs. In experiment 1, owners collected voided urine samples from 19 healthy pet dogs at specified times before and after taking the dogs to a veterinary practice for yearly vaccination. In experiment 2, 12 pet dogs were evaluated in a similar way before and after an orthopedic examination at a referral clinic. In experiment 3, 9 healthy pet dogs were hospitalized for 1.5 days and urine samples were collected before, during, and after this stay. Basal urinary C:C ratios in all experiments ranged from 0.8 to 8.3 x 10(-6). In experiment 1, the urinary C:C ratio after the visit to the veterinary practice ranged from 0.9 to 22.0 x 10( 6). Six dogs had a significantly increased urinary C:C ratio (responders), but in 5 of these dogs the ratio was < or = 10 x 10(-6). In experiment 2, 8 of 12 dogs responded significantly with urinary C:C ratios ranging from 3.1 to 27.0 x 10( 6). In experiment 3, 8 of 9 dogs had significantly increased urinary C:C ratios, ranging from 2.4 to 24.0 x 10(-6), in some or all urine samples collected during hospitalization. In 4 dogs urinary C:C ratios 12 hours after hospitalization were still significantly higher than the initial values. Thus, a visit to a veterinary practice, an orthopedic examination in a referral clinic, and hospitalization can be considered stressful conditions for dogs. A large variation occurs in response, and in individual dogs the increases in urinary C:C ratios can exceed the cutoff level for the diagnosis of hyperadrenocorticism. Therefore, urine samples for measurement of the C:C ratio in the diagnosis of hyperadrenocorticism should be collected in the dog's home environment, to avoid the influence of stress on glucocorticoid secretion. PMID- 9857336 TI - Survival in dogs with nasal adenocarcinoma: 64 cases (1981-1995). AB - Case records of 64 dogs with nasal adenocarcinoma were reviewed. The effects of age, gender, tumor stage, presence of metastatic lesions, and treatment method on survival time were examined. Surgery groups included rhinotomy (n = 9), transnasal curettage (n = 29), and no surgery (n = 26). Chemotherapy groups included fluorouracil-cyclophosphamide combination therapy (n = 15), mitoxantrone (n = 7), and no chemotherapy (n = 42). Fifty-three dogs received fractionated cobalt 60 radiation therapy. Surgical procedure, chemotherapy group, and stage of primary tumor were not significantly associated with survival time (P > .05). Dogs that received radiation therapy had a significantly longer median survival time (424 days) than dogs that did not (126 days)(P = .0001). The presence of either regional lymph node or pulmonary metastasis was associated with significantly shorter median survival time (109 days) when compared to dogs without metastases (393 days)(P = .0125). When only dogs that had received radiation therapy were considered, neither surgical treatment nor chemotherapy group was associated with significant changes in median survival time. An alternate staging system emphasizing the presence or absence of metastases is proposed. PMID- 9857338 TI - Treatment of neonatal calf diarrhea with an oral electrolyte solution supplemented with psyllium mucilloid. AB - Dairy calves under 14 days of age with naturally occurring, uncomplicated diarrhea were treated for 3 days with a hypertonic oral electrolyte solution with (n = 15) or without (n = 12) psyllium. Clinical response and clinical pathology data were compared between the 2 groups. Glucose absorption was evaluated on days 1 and 3 by measurement of plasma glucose and lactate and serum insulin concentrations for 4 hours after formula administration. On day 1, glucose, lactate, and insulin concentrations were lower in psyllium-fed calves than in control calves, with significant differences noted in glucose and lactate concentrations at several time points (P < 0.05). Plasma lactate concentrations were higher at several times in both treatment groups on day 3 than on day 1 (P < 0.05). Fecal consistency was markedly different in psyllium-fed calves as compared with control calves within 24 hours of psyllium supplementation. Fecal percent dry matter content was lower in psyllium-fed calves than in control calves at least once a day during supplementation and on day 3 compared with day 0 in the psyllium-fed calves (P < 0.05). There were no significant differences in clinical performance scores, hydration status, arterial blood gas, serum anion gap, electrolyte, or total CO2 concentrations. Addition of psyllium to an oral electrolyte solution resulted in immediate alterations in glucose absorption without impairing rehydration in diarrheic calves, but differences were transient and did not affect clinical outcome. PMID- 9857339 TI - Survey of the large animal diplomates of the American College of Veterinary Internal Medicine regarding percutaneous lung biopsy in the horse. AB - A survey designed to obtain information on the indications, contraindications, complications, and methodology of percutaneous lung biopsy in the horse was sent to large animal diplomates of the American College of Veterinary Internal Medicine. Sixty-five of 190 diplomates returned the survey (response rate: 34%) and 59 of these 65 respondents (91%) indicated that they worked with horses. Forty-four diplomates had performed a percutaneous lung biopsy in 1 or more horses (i.e. 75% of those diplomates working with horses and 68% of total respondents). Clinical and radiologic diagnoses that prompted diplomates to perform percutaneous lung biopsy in the horse included a pulmonary miliary pattern (93%), suspicion of pulmonary infiltrative disease (91%), suspicion of pulmonary neoplasia (91%), suspicion of chronic obstructive pulmonary disease (COPD) (20%), and suspicion of exercise-induced pulmonary hemorrhage (EIPH) (7%). Only one of the respondents reported the use of percutaneous lung biopsy in the diagnostic workup if pneumonia was suspected, but 11% of respondents reported that suspicion of pulmonary abscessation would prompt them to perform a percutaneous lung biopsy. In contrast, a variable percentage of respondents felt there were contraindications to performance of this technique, which included neonatal septicemia (68%), pulmonary abscessation (65%), pleuropneumonia (55%) and pneumonia (42%), EIPH (41%), and COPD (26%). No respondent indicated that suspicion of neoplasia was a contraindication to percutaneous biopsy. Most common complications observed by respondents were epistaxis (68% of respondents), putative pulmonary hemorrhage (52%), tachypnea (39%), and respiratory distress (32%). Ten of 44 respondents (23%) had not seen any complications with percutaneous lung biopsy. Forty-two of 44 respondents (96%) warned owners about possible complications before performing percutaneous lung biopsy. All respondents to this question reported that they would perform percutaneous lung biopsies in horses in the future, but 4 of 41 would use the procedure only as a last resort. PMID- 9857337 TI - Nutritional alterations and the effect of fish oil supplementation in dogs with heart failure. AB - Alterations in body composition and nutritional status are common in humans with heart failure and are related, in part, to increases in cytokine concentrations. Cytokines have not been studied previously in dogs with naturally occurring cardiac disease nor has fish oil administration been used in this population to decrease cytokine production. The purposes of this study were to characterize nutritional and cytokine alterations in dogs with heart failure and to test the ability of fish oil to reduce cytokines and improve clinical outcome. Body composition, insulinlike growth factor-1, fatty acids, and cytokines were measured in 28 dogs with heart failure and in 5 healthy controls. Dogs with heart failure then were randomized to receive either fish oil or placebo for 8 weeks. All parameters were measured again at the end of the study period. At baseline, 54% of dogs with heart failure were cachectic and the severity of cachexia correlated with circulating tumor necrosis factor-alpha concentrations (P = .05). Cytokine concentrations at baseline, however, were not significantly increased in dogs with heart failure compared to controls. Baseline plasma arachidonic acid (P = .02), eicosapentaenoic acid (P = .03), and docosahexaenoic acid (P = .004) concentrations were lower in dogs with heart failure than in controls. Fish oil supplementation decreased interleukin-1 beta (IL-1) concentrations (P = .02) and improved cachexia (P = .01) compared to the placebo group. The mean caloric intake of the heart failure dogs as a group was below the maintenance energy requirement (P < .001), but no difference was found in food intake between the fish oil and placebo groups. Insulinlike growth factor-1 concentrations (P = .01) and reductions in circulating IL-1 concentrations over the study period (P = .02) correlated with survival. These data demonstrate that canine heart failure is associated with cachexia, alterations in fatty acids, and reduced caloric intake. Fish oil supplementation decreased IL-1 concentrations and improved cachexia. In addition, reductions in IL-1 predicted survival, suggesting that anticytokine strategies may benefit patients with heart failure. PMID- 9857340 TI - A combination chemotherapy protocol (VELCAP-L) for dogs with lymphoma. AB - Ninety-eight dogs with lymphoma treated with a 5-drug combination chemotherapy regimen (vincristine, L-asparaginase, cyclophosphamide, doxorubicin, prednisone [VELCAP-L]) were evaluated for pretreatment characteristics predictive for response and remission duration. The complete remission rate was 69%, with a median remission duration of 55 weeks. Dogs with advanced stage of disease, constitutional signs, dogs that were older, and dogs that were dyspneic were less likely to achieve remission. Once in remission, small dogs and dogs without pretreatment thrombocytopenia were likely to have longer remission duration. Toxicoses were frequent, but rarely fatal, and no predictitive factors were found for a dog developing toxicoses. VELCAP-L is an effective treatment for dogs in stage I-III lymphoma, particularly in young, small animals. PMID- 9857341 TI - Systemic plasmacytosis and polyclonal gammopathy in a dog. PMID- 9857343 TI - "Comparison of some pharmacokinetic parameters of 5 angiotensin-converting enzyme inhibitors in normal beagles". PMID- 9857342 TI - Hyperthyroidism associated with a thyroid adenocarcinoma in a 21-year-old gelding. PMID- 9857344 TI - Abnormalities of the thoracic bellows: stress fractures of the ribs and hiatal hernia. PMID- 9857345 TI - Structure and function of the encapsidation signal of hepadnaviridae. AB - The hepatitis B virus (HBV) and other members of the hepadnaviridae replicate by reverse transcription of an RNA intermediate, pregenomic RNA (pgRNA). pgRNA is also translated into core protein and polymerase (reverse transcriptase) protein. Before being reverse transcribed, pgRNA is sequestrated from the cytoplasm by being packaged, together with polymerase, into subviral particles composed of core protein. For pgRNA to be encapsidated, its 5' end is folded into a stem-loop structure, known as the encapsidation signal or epsilon (epsilon). This stable bipartite stem-loop structure contains a bulge and an apical loop. Besides encapsidation, epsilon is involved in the activation of polymerase, in template restriction and in the initiation of DNA synthesis by reverse transcription. HBV DNA encoding epsilon forms part of the template that is translated into the precore/core fusion protein that is in turn post-translationally modified to produce hepatitis B e antigen (HBeAg). The DNA encoding epsilon may be recombinogenic. Mutations within epsilon can affect its function and sequence conservation within epsilon in natural isolates is therefore high. epsilon could provide a practical target for antiviral therapy. PMID- 9857346 TI - Liver-directed gene transfer: a linear polyethlenimine derivative mediates highly efficient DNA delivery to primary hepatocytes in vitro and in vivo. AB - Efficient DNA delivery is a prerequisite for the successful implementation of molecular antiviral strategies against chronic viral hepatitis and gene therapy in general. The cationic polymer polyethylenimine (PEI) has recently been explored as a gene transfer vector in various cell types in vitro and in vivo. In this study, we evaluated a linear PEI derivative (lPEI) as a vector for gene and oligodeoxynucleotide transfer into hepatocytes in vitro and in vivo. A simple protocol was developed that allowed transfection of up to 50% of primary hepatocytes in vitro. In addition, fluorescent oligodeoxynucleotides were efficiently delivered to the liver in vivo after intravenous injection into Pekin ducks. Thus, lPEI mediates highly efficient gene and oligodeoxynucleotide transfer into primary hepatocytes and is potentially useful for DNA delivery in vivo. PMID- 9857347 TI - Hepatitis B virus infection in microcarrier-attached immortalized human hepatocytes cultured in molecularporous membrane bags: a model for long-term episomal replication of HBV. AB - Studies on the pathobiology of chronic (long-term) hepatitis B virus (HBV) infection and in vitro drug testing have been hampered by the lack of appropriate systems for culturing susceptible cells chronically infected with HBV. Most of the in vitro studies of HBV replication have been performed with HBV genome transduced cell lines. In this system, viral production is mainly the result of chromosomal replication. In an in vitro infection system, owing to medium refreshment (which leads to the removal of infectious particles necessary for the perpetuation of infection) and to trypsinization for cell passages, it is difficult, if not impossible, to maintain chronic HBV infection, despite the use of susceptible cells. To circumvent these unfavourable factors for chronic HBV infection in vitro, we cultured microcarrier-attached immortalized human hepatocytes, infected with HBV, in molecularporous (MW 12,000-14,000) membrane (dialysis) bags for a duration of 2 months. HBV covalently-closed-circular (ccc) DNA, HBV precore/core and X mRNAs were detected in the cells cultured in this system following infection until the end of the experiment (day 58), while in classical culture conditions (monolayer), markers of HBV replication were also detected. Production of hepatitis B surface antigen (HBsAg) and HBV DNA was detected and their levels in culture medium (collected at the end of experiments from the molecularporous membrane bags) were increased 2.86- and 3.28-fold respectively. Using Southern blot analysis, HBV replicative intermediates could also be demonstrated throughout the experiments. However, integrated HBV DNA was not present. In contrast, HBV ccc DNA, HBV precore/core and X mRNAs, and replicative intermediates were not demonstrable in FTO 2B rat hepatoma cells infected in the same manner in parallel experiments. This in vitro infection system, using susceptible, immortalized human hepatocytes, therefore provides a new tool for studying the long-term effect of HBV infection, mainly involving episomal replication in hepatocytes, and for drug testing. PMID- 9857348 TI - The long-term effect of treatment with interferon-alpha 2a in chronic hepatitis B. The Long-Term Follow-up Investigator Group. The European Study Group on Viral Hepatitis (EUROHEP). Executive Team on Anti-Viral Treatment. AB - This study was performed to evaluate the long-term effects of interferon-alpha 2a (IFN-alpha 2a) vs no treatment in patients with chronic hepatitis B and to determine whether viral clearance, following therapy or occurring spontaneously, was sustained. Patients originating from three previously published multicentre, randomized, controlled trials were analysed. Information about survival and response during long-term follow-up was available in 340 (73%) and 308 (66%) of 469 randomized patients respectively. Response to therapy (viral clearance) was defined as: loss of hepatitis B virus (HBV) DNA and loss of hepatitis B e antigen (HBeAg) and improvement in alanine aminotransferase level. Scheduled treatment free follow-up was 12 months in all studies. Median long-term follow-up time after inclusion in the individual studies was 4.7 years (range: 0.2-7.5 years). Viral clearance after IFN-alpha 2a, or occurring spontaneously, was sustained in 70 out of 80 evaluable patients (88%) who were responders at the end of the original trials and 21 (30%) lost hepatitis B surface antigen (HBsAg). A total of 80 patients received (re)treatment during the long-term follow-up period and 33% of them responded, irrespective of previous treatment category. Overall response rate was not significantly affected by gender, sexual inclination or ethnic origin. Durability of response did not depend upon ethnic origin or presence of cirrhosis. At the end of the original trial periods, 253 patients were histologically evaluated and 22 (9%) had histologically confirmed progression to cirrhosis. During long-term follow-up an additional five patients developed cirrhosis. Hepatocellular carcinoma developed in three patients (1%): in one patient during the follow-up period of the original trial and in two patients (one untreated) during the long-term follow-up period. Ten of 25 deaths were liver-related (hepatocellular carcinoma in three, gastrointestinal bleeding in two and liver failure in five). The distribution of clinical events (progression to cirrhosis, hepatocellular carcinoma and liver-related deaths) was unrelated to original treatment category and response to treatment. Hence, 90% of responding patients will, irrespective of treatment category, have a sustained response. At least 30% of responding patients will eventually lose HBsAg. For a number of reasons, the present patient population and observation period are insufficient to establish a presumed beneficial effect of IFN-alpha 2a on disease progression and survival. PMID- 9857349 TI - Viral and host factors in the prediction of response to interferon-alpha therapy in chronic hepatitis C after long-term follow-up. AB - Acute infection with hepatitis C virus (HCV) develops into a chronic hepatitis in about 50-70% of patients. Treatment of these patients with interferon-alpha (IFN alpha) results in a sustained long-term response in only 15-20% but causes numerous unwanted side-effects in a higher percentage of patients. The aim of our study was to define host or viral parameters that would allow identification of responders and non-responders to IFN-alpha prior to the onset of treatment. We studied a group of 87 patients suffering from chronic hepatitis C who were treated with IFN-alpha. After long-term follow-up, 18 patients (21%) showed a sustained response to IFN-alpha therapy (normalization of serum transaminases and loss of viral RNA from serum) for up to 7 years after therapy had ceased. By univariate and multivariate analyses, no host factors were found to be predictive of response to therapy. Neither the degree of inflammation or fibrosis in liver biopsy samples obtained before treatment nor immunogenetic factors (major histocompatibility complex II haplotype and tumour necrosis factor-alpha promoter polymorphism) were associated with response to therapy. In contrast, viral parameters showed a strong association with response to therapy. HCV genotype 3 was found significantly more frequently in responders (P = 0.034), and mean HCV RNA concentration was lower in responders (3.1 x 10(4)) than in non-responders (2.5 x 10(5)) (P = 0.01). By multivariate analysis, both HCV genotype and HCV RNA concentration were independent predictors of response to therapy. However, exact prediction of response to treatment for an individual patient was not possible on the basis of pretreatment viral RNA concentration or viral genotype. The best association with response to therapy was found to be clearance of HCV RNA from serum 3 months after the start of treatment (32 of 34 partial and sustained responders vs 0 of 53 non-responders; P = 0.001). In conclusion, determination of pretreatment viral factors, but not host factors, was significantly correlated with treatment response but did not give an accurate prediction for patients, whereas clearance of HCV RNA from serum after 3 months of therapy was predictive of response to therapy. PMID- 9857350 TI - Clinical evaluation of the branched DNA assay for hepatitis B virus DNA detection in patients with chronic hepatitis B lacking hepatitis B e antigen and treated with interferon-alpha. AB - The aim of this study was to evaluate the Chiron branched DNA (bDNA) assay for detection of serum hepatitis B virus (HBV) DNA in patients with chronic hepatitis B lacking hepatitis B e antigen (HBeAg) and undergoing interferon (IFN) therapy. Results obtained with the bDNA assay were compared with those obtained using the Abbott liquid hybridization (LH) assay and the polymerase chain reaction (PCR). Serial samples (274) from 34 patients were analysed. Analysis of variance results indicated that bDNA values were more significantly correlated than LH values with both PCR positive/negative results (probability of artifact (Prob > F) = 0.7 and 0.09 for LH and bDNA assays, respectively) and presence/absence of precore mutations (Prob > F = 0.21 and 0.001 for LH and bDNA assays, respectively). Both bDNA and LH results correlated highly with alanine aminotransferase (ALT) values (both had Prob > F values of 0.0) while PCR was not correlated with ALT (Prob > F = 0.05). In 26 evaluable patients, a model based on a generalized Knodell score was used to predict response to IFN therapy, as defined by normalization of ALT values during therapy. This model discriminated well between non-responders and responders. The bDNA results correlated well with the generalized Knodell score, while the LH results did not (Prob > F = 0.04 and 0.19 for the bDNA and LH assays, respectively). In conclusion, the bDNA assay appears to be useful for quantification of HBV DNA levels in HBeAg-negative chronic hepatitis as it correlates with biochemical and histological indications of disease severity as well as with response to IFN therapy. PMID- 9857351 TI - Multi-measurement method comparison of three commercial hepatitis B virus DNA quantification assays. AB - Serum specimens (327) from patients with chronic hepatitis B were evaluated for hepatitis B virus (HBV) DNA using three commercial assays--Chiron Quantiplex (CA), Digene Hybrid Capture (DA) and Abbott HBV DNA assay (AA). The HBV DNA values obtained following evaluation were used to compare the linearity, responsiveness and precision of each assay and to determine the conversion between the three different assay values. The comparison was accomplished using a new statistical approach termed the multi-measurement method (MMM). MMM is a minimal bias, non-linear regression technique that allows simultaneous multiassay performance evaluation as well as assay value interconversion. MMM analysis demonstrated that the CA was more sensitive and responsive than either the DA or the AA. Both the CA and DA were more precise than the AA. Validation of the MMM results was performed using two additional data sets of 551 and 100 specimens, respectively. MMM is a novel statistical tool that has a broad application for the generation of statistically normalized laboratory data and for interassay standardization. PMID- 9857352 TI - Evidence for the genetic basis of non-response in mice. AB - In the murine model the immune response to both the hepatitis B envelope and nucleocapsid antigens are linked to the MHC class II haplotype but they are independently restricted. Although in a human outbred population non-response is less likely, the host genetic environment may be important in determining the level of response to vaccine and inclusion of antigens apart from the S antigen may improve protection. The inclusion of HBcAg in vaccine remains controversial since, despite its high immunogenicity, the Th-cell response to this antigen may have a role in the down regulation of the immune response to HBV so promoting persistence. PMID- 9857353 TI - Characterization of the T- and B-cell immune response to a new recombinant pre S1, pre-S2 and SHBs antigen containing hepatitis B vaccine (Hepagene); evidence for superior anti-SHBs antibody induction in responder mice. AB - Hepagene is a novel recombinant particle consisting of the pre-S1, pre-S2 and small surface (SHBs) antigens (Ag) of the hepatitis B virus (HBV) and is adjuvanted with alhydrogel in the final formulation. It has been primarily developed to enhance anti-SHBs antibody titres in inadequate responders, to conventional SHBsAg vaccines. Since non-compliance is also a problem with existing HBV vaccine schedules, the ability to accelerate current immunization regimens to provide more rapid protection has also been an important objective. Here we describe the T- and B-cell responses to Hepagene in two strains of responder mouse (BALB/c and SWR/J). Hepagene induced high in vitro spleen T-cell proliferative responses in both strains (max. Stimulation Index = 43), following intraperitoneal immunization. High concentrations of interferon-gamma (max. = 5000 pg/mL) were detected in the media of spleen cells cultured with non adjuvanted Hepagene particles. SWR/J mice showed the highest serum antibody (Ab) titres to non-adjuvanted Hepagene. The presence of alhydrogel adjuvant in the vaccine formulation significantly improved the titres. Anti-pre-S1 Ab was detected in both strains of mouse but anti-pre-S2 Ab was only detected in the SWR/J strain. In BALB/c mice, the anti-Hepagene (non-adjuvanted) IgG1 Ab subclass was predominant but in SWR/J mice IgG1, IgG2a and IgG2b subclasses were of a similar magnitude. In BALB/c mice, Hepagene induced higher anti-SHBs Ab responses than Engerix-B (11097 IU/mL and 1276 IU/mL, respectively), following two doses of vaccine (10 micrograms/mouse). The vaccine therefore, induces strong cellular and humoral immune responses and these data suggest that Hepagene is an improved hepatitis B vaccine. PMID- 9857354 TI - Hepatitis B third-generation vaccines: improved response and conventional vaccine non-response--evidence for genetic basis in humans. AB - The lack of response to hepatitis B vaccination remains a problem for those individuals directly at risk of hepatitis B infection, particularly those who work in the health care industry. The factors associated with non-response to hepatitis B vaccination have been investigated in 86 non-responder health care workers who had received multiple 'S' vaccinations without sustained production of anti-HBs. This group received a recently developed hepatitis B vaccine, Hepagene, which included proteins derived from the envelope region of HBV, not present in currently licensed vaccines. The pre-S1 and pre-S2 proteins were included in Hepagene in order to circumvent anti-HBs non-responsiveness which had previously been demonstrated in the inbred mouse model. The inclusion of these additional proteins in Hepagene enabled some seroconverion, from non-responder to responder; however, a proportion of the vaccinees remained non-responders and the reasons for this have been investigated here, with reference to HLA alleles and the demographic predisposition. Here the mechanisms that underlie hepatitis B vaccine non-response have considered the distribution of HLA alleles, age, sex, height and weight in addition to the T-cell responses to Hepagene derived antigens. PMID- 9857355 TI - Hepatitis B third-generation vaccines: improved response and conventional vaccine non-response--third generation pre-S/S vaccines overcome non-response. AB - A new triple-S containing recombinant hepatitis B vaccine was evaluated in terms of immunogenicity and reactogenicity in a cohort of healthy healthcare professionals who were persistent non-responders to the currently licensed hepatitis B vaccines. One hundred subjects were allocated randomly to receive two doses of 5, 10, 20 or 40 micrograms of a new hepatitis B vaccine 2 months apart. The overall seroconversion rate was 70% with a single dose of 20 micrograms of the vaccine being as effective as two doses of either 20 micrograms or 40 micrograms of the vaccine formulation in terms of seroconversion, seroprotection and geometric mean titres. PMID- 9857356 TI - Current aspects of hepatitis B surface antigen mutants in Singapore. AB - Mutations occurring on the major antigenic 'a' determinant of hepatitis B surface antigen (HBsAg) have been detected in immunized Singapore children, despite immunoprophylaxis with hepatitis B immune globulin and HBV vaccine. These vaccine escape HBsAg mutants display a predominance of the Gly145-to-Arg145 mutation in the antigenic 'a' determinant. Our latest follow-up studies indicate the stability of this as well as other vaccine-escape HBsAg mutants over time. We have also identified HBsAg mutants in immunized children with amino acid substitutions outside the 'a' determinant. Transmission of various vaccine-escape HBsAg mutants have been shown in our epidemiological studies. Detection of HBsAg mutants, carrying amino acid changes at various position of the 'a' determinant in random population, points to their emergence in Singapore before the implementation of the vaccination programme. Of significant interest is the recent identification of vaccine-escape HBsAg mutants in hepatocellular carcinoma. PMID- 9857358 TI - Mechanism of emergence of hepatitis B virus escape variants: approaches to prevention. AB - Hepatitis B virus causes a chronic infection in a large proportion of cases. One possible mechanism whereby hepatitis B avoids the host defences is by mutation of antigenic epitopes. This article reviews current views in the area. PMID- 9857357 TI - Current status of HBV vaccine escape variants--a mathematical model of their epidemiology. AB - We present a deterministic model of the possible emergence of a vaccine escape variant of hepatitis B virus (HBV). The model identifies the key unknowns determining this process: the protection afforded by the current vaccines against particular HBV variants; the infectiousness of these variants; and the current prevalence of individuals infectious with the variants (each factor relative to wild-type). By making pessimistic assumptions about these unknowns we show that even a highly infectious variant, under a vaccine programme that affords no protection against the variant, would still take decades to emerge. Thus the current low prevalence of variants is not evidence for the cross-reactivity of the current vaccines or for a lack of infectiousness in the variants. As any vaccine failure will be inapparent for decades it may be sensible to recommend vaccine modifications now rather than later. PMID- 9857359 TI - New US cancer genetics network announced. PMID- 9857360 TI - Towards the biosynthesis of an alternative to Taxol. PMID- 9857361 TI - Muscling in on atherosclerosis. PMID- 9857362 TI - Protease inhibitors on trial for inflammatory bowel disease. PMID- 9857363 TI - Tolerizing B cells to jump the first hurdle in xenotransplantation. PMID- 9857364 TI - Ageing research in the UK: plenty of proliferative potential. Cellular Senescence: the future of ageing. Oxford, United Kingdom, 16-17 July 1998. PMID- 9857365 TI - Germline alteration by gene therapy: assessing and reducing the risks. AB - Developments in gene therapy are certain to lead to the treatment of an increasing variety of diseases, some of which will affect patients who might wish to have children following their gene therapy treatment. These circumstances raise the concern that germline integration of gene therapy vector DNA could occur. Although our current understanding of reproductive biology and of the biodistribution of gene therapy vectors administered to extragonadal sites indicate that this risk is low, animal experiments and clinical studies designed specifically to address this question are warranted; because of this risk, every gene therapy vector should be tested for its potential to integrate into germ cells and preimplantation embryos. PMID- 9857366 TI - Sertoli cell transplants: their use in the treatment of neurodegenerative disease. AB - The efficacy of treating neurodegenerative diseases with the transplantation of fetal tissue has been demonstrated in animal models of Parkinson's disease, Huntington's disease and stroke. In the clinical setting, neural transplantation as a treatment for patients with Parkinson's disease has shown promising results. However, for this treatment method to be effective neuronal survival needs to be improved through either trophic support or localized immunoprotection. Co transplanting Sertoli cells, which express many nutritive, regulatory, trophic and immunosuppressive factors, with fetal neural cells could provide both of these requirements. Such a strategy could enhance the recovery benefits associated with transplantation and decrease the need for, and the risks associated with, long-term systemic immunosuppression. PMID- 9857367 TI - Heat shock protein--peptide complexes as immunotherapy for human cancer. AB - Heat shock proteins (Hsps), ubiquitous in nature, act as chaperones for peptides and other proteins. They have been implicated in loading immunogenic peptides onto major histocompatibility complex molecules for presentation to T cells. When isolated from tumor cells, Hsps are complexed with a wide array of peptides, some of which serve as tumor-specific antigens. Animal studies have demonstrated that heat shock protein--peptide complexes (HSPPCs) from tumor cells can act as vaccines to prevent or treat tumors. Potent and specific tumor antigens have long been the holy grail in cancer immunotherapy; HSPPCs from tumor cells could become a safe and reliable source of tumor-specific antigens for clinical application. PMID- 9857368 TI - Towards gene therapy for the central nervous system. AB - Gene therapy has generated enormous scientific, medical and public interest over the last decade. Clinical trials involving approximately 2000 patients worldwide have targeted simple genetic diseases such as cystic fibrosis, muscular dystrophy, adenosine deaminase deficiency, Gaucher's disease and familial hypercholesterolemia, as well as complex acquired diseases such as cancer and AIDS. The central nervous system is a new and particularly exciting target for gene therapy because its unique properties prevent the successful treatment of many neurological disorders by conventional means. This review discusses the potential applications of in vivo gene therapy to neurological disorders that have the greatest potential for genetic treatments. PMID- 9857369 TI - Prospects for gene therapy in human prostate cancer. AB - Prostate cancer is the most common neoplasm in men and a significant cause of mortality in affected patients. Despite significant advances, current methods of treatment are effective only in the absence of metastatic disease. Gene therapy offers a renewed hope of using the differential characteristics of normal and malignant tissue in constructing treatment strategies. Several clinical trials in prostate cancer gene therapy are currently under way, using immunomodulatory genes, anti-oncogenes, tumor suppressor genes and suicide genes. A continued understanding of the etiological mechanisms involved in the establishment and progression of prostate cancer, along with advances in gene therapy technology, should make gene therapy for prostate cancer therapeutically valuable in the future. PMID- 9857370 TI - [Physiopathology of the renin-angiotensin system in the kidney. Proceedings of a meeting. Le Coudray Montceaux, 27-28 March 1998]. PMID- 9857371 TI - [Polymorphism of the renin-angiotensin-aldosterone system: molecular and epidemiogenetic aspects]. AB - The involvement of polymorphic variations in the structure of the renin angiotensin aldosterone system genes in the predisposition to cardiovascular disease is a particularly important field of investigation. A large number of studies have been published and their results are often contradictory. It is therefore important to identify the functional variants modifying the level of expression of these genes or the structure of the encoded proteins. This will allow the clear quantitation of their effect on quantitative phenotypes both in vitro and in vivo. It is also important to study in the future new phenotypes, particularly dynamic phenotypes, potentially influenced by these genotypes and to use them for building a new cardio-vascular pharmacogenetics. PMID- 9857372 TI - [Experimental genetic modifications in the renin-angiotensin and kallikrein-kinin systems: significance for understanding cardiovascular regulation and study of genetic determinism of human diseases]. AB - The precise role of the different proteins that constitute the renin-angiotensin and kallikrein-kinin systems in the development of hypertension and some cardiac and renal diseases remains unclear. Genetic manipulations in animals is a powerful approach that provide the opportunity to explore the role of each of these proteins in vivo. Indeed it is possible in the rat and in the mouse to manipulate a specific gene without modifying the other genetic and environmental factors. A causal link can thus be established between the gene and a physiologic or pathologic alteration. The possibilities are either the overexpression of the gene in all or in specific tissues (transgenesis), or the modification (often the inactivation) of the endogenous gene by homologous recombination. The second technique has the advantage to be more specific but it can be used only in the mouse; it is performed by transfecting totipotent embryonic stem cells with a vector harboring identical sequences to those of the gene to be targeted. The embryonic stem cells are then injected into embryos in which they will participate in the generation of the different organs including the gonads. The resulting chimeric animals can therefore transmit the mutation to their offspring creating a new genetically modified mouse strain. Many strains targeted in the different components of the renin-angiotensin and kallikrein-kinin systems have been generated in this way. These animal models should allow to test many physiopathological hypotheses that have been put forward from the results of human genetics and clinical studies, and also to raise new ones. PMID- 9857373 TI - Examining the renin-angiotensin system one hundred years after its discovery. AB - This article reviews the biology of angiotensin converting enzyme (ACE). There are two ACE isozyme, somatic ACE (made by tissues such as the lung and kidney) and testis ACE (produced only by developing male germ cells). Mice lacking all ACE expression were prepared using homologous recombination of embryonic stem cells. These animals have profoundly low blood pressures, reduced male fertility and a renal lesion characterized by under development of the renal medulla and papilla. A second line of mice was made in which ACE activity is found in the plasma but is absent from all tissues such as lung. These animals have a phenotype very similar to mice lacking all ACE with the exception of the renal lesion which is much less pronounced. This second line of mice strongly suggests that it is tissue-bound ACE which is critical in the proper functioning of the renin-angiotensin system. PMID- 9857374 TI - Towards gene therapy for renal diseases. AB - The rationale of the somatic gene therapy is the correction of diseases at the most fundamental level. Ideal gene therapy should be achieved by the replacement of the wrong gene sequence of genome with correct one. However, the gene technology to date is yet immature so as to correct the wrong gene sequence in vivo. Potentially, the present technology of gene transfer may provide: 1) correction of cellular dysfunction by expressing the deficient gene; 2) addition of new function for a cell by transferring an exogenous gene; 3) inhibition of unfavorable action of a cell by introducing a counteracting gene. In nephrology, the gene transfer targeted kidney has been challenged at the experimental level. HVJ-liposome method and recombinant adenovirus allow gene transfer to the particular cells in kidney in vivo. Ex vivo gene transfer using mesangial cells and macrophages are another option. Transplant kidney is also a good material for genetic engineering. The potential application of gene transfer is enormous while the therapeutic application have just begun to explored. We have been devoted to HVJ-liposome mediated gene transfer to the kidney and successfully demonstrated the suppression of the extracellular matrix accumulation of the glomeruli in the experimental glomerulonephritis through inhibition of the TGF-beta action by antisense oligonucleotides or soluble type receptor chimera for TGF-beta. We also applied this technology to the inhibition of interstitial fibrosis in unilateral ureter obstruction model. The new HVJ-liposome method improved in lipid composition allows gene transfer to tubulointerstitial fibroblast by retrograde approach from ureter. In consequence, introduced TGF-beta antisense suppressed the TGF-beta mRNA in concomitant with ameliorating interstitial fibrosis. We believe that the gene transfer technique will become common strategy to study the molecular aspect of the renal diseases and will be possibly applicable to molecular intervention in nephrology. PMID- 9857375 TI - [Molecular structure and function of angiotensin ii receptors]. AB - Angiotensin II (Ang II) receptors are 7 transmembrane domain receptors corresponding to 2 pharmacologically and molecularly distinct receptors, called AT1 and AT2, the primary structures of which have been established by molecular cloning. Most if not all the physiological actions of Ang II are mediated by the AT1 receptor, which is coupled to a Gq protein activating a phospholipase C (PLC), which in turn mobilizes the intracellular calcium stores and activates protein kinases C. Many site directed mutagenesis works have allowed to identify short extracellular sequences responsible for the Ang II binding, whereas non peptidic AT1-specific antagonists bind to a different transmembranar site. Structural modifications are responsible for the change of the receptor from an inactive to an active state. At the basal state, the receptor is mostly in an inactive state; agonists present a better affinity for the active state, displacing the equilibrium to this state; at the opposite, the inverse agonists present a better affinity for the inactive state. Antagonists present a similar affinity for both states of the receptor. Several mutations of polar residues of the transmembrane domains block the receptor either in an inactive state (D74D, S115A, Y292F) or in a constitutively active state (N111A and N295A). After activation, the receptor is coupled to different intracellular proteins, the first of them being the G proteins of the Gq/11 family. The sequences of the receptor involved in this coupling correspond to the 2nd, the 3rd intracellular loops and the proximal segment of the carboxyterminal domain. Other sequences interact with other proteins, such as the 319YIPP332 sequence of the carboxyterminus, which interacts with the Jak2 tyrosine kinase. After the binding of a peptidic ligands, the ligand-receptor complex is internalized independently for the G protein coupling. Again, site directed mutagenesis experiments have localized a sequence of the carboxyterminus (329STLSTKMSTLS338) involved in the internalization. This serine and threonine-rich sequence plays also a role in the desensitization of the AT1 receptor, consecutively to its phosphorylation. The AT2 receptor is only 34% identical to the AT1 receptor and its functions are far less understood. Its physiological functions (apoptosis and antiproliferative actions) and its signaling pathways (activation of Gi proteins and tyrosine phosphatases) are still a matter of debate. PMID- 9857376 TI - [Evidence of a renin receptor on human mesangial cells: effects on PAI1 and cGMP]. AB - Most proteases a receptor or a binding site that serves to concentrate the proteolytic activity on the cell surface and to mediate cellular effects. We looked for such a receptor for renin, an aspartyl protease. The binding of recombinant human renin labelled with 125I was studied on primary and immortalized human mesangial cells. The binding of renin was specific, saturable and was characterized by Kd = 0.4 nM and 8,000 sites/cell and Kd = 1 nM and 2,000 sites/cell for primary and immortalized cells, respectively. The binding did not depend on the active site of the enzyme, was not followed by internalization and degradation of renin and did not modify intracellular Ca2+. Stimulation of primary cells with 100 nM induced a significant increase of 3H thymidine incorporation but was not associated with an increase of the cell number. Furthermore, incubation of mesangial cells 24 h with 100 nM renin provoked an increase of tPA and of PAI1 in the conditioned medium. This increase was not modified neither by captopril nor by angiotensin II receptors antagonists. The tPA antigen elevation was confirmed by fibrin zymography showing an increase of tPA/PAI1 complexes. But, surprisingly, the reverse zymogram showed that PAI antigen increase was associated with decreased PAI activity which was due to PAI clivage in an inactive form. PAI clivage by renin required the presence of the cells and could not be obtained by incubating renin and recombinant human PAI alone. When primary mesangial cells were cultured in the presence of a specific inhibitor of renin active site, RO 42-5982, PAI accumulation in the conditioned medium was reduced by 50-60%, suggesting that endogenous renin plays a role in PAI synthesis and/or secretion. The binding of renin does not induce cAMP and cGMP generation. However, in the presence of renin (100 nM and 1 microM) the extent of cGMP generated by CNP (10 and 100 nM) was reduced by 50%. Preliminary results of the renin receptor purification by affinity chromatography indicate that the receptor Mr is about 57 kDa. PMID- 9857377 TI - Identification of a novel dual angiotensin II/vasopressin receptor. AB - The isolation and molecular characterization of the Ang II/AVP receptor elucidates the structure of a novel dual receptor coupled to adenylate cyclase and responding with equal sensitivity to Ang II and AVP. The cloning strategy in conjunction with site directed mutagenesis have permitted the delineation of the Ang II and AVP binding domains within the receptor polypeptide. Pharmacological characterization of the receptor defines the AngII/AVP receptor as a novel AT1/V2 type of receptor. The renal immunocytochemical distribution of the Ang II/AVP receptor to the outer medullary thick ascending limb tubules and inner medullary collecting ducts suggests a prominent role in renal tubular sodium and fluid reabsorption. PMID- 9857378 TI - [Effect of angiotensin ii on Na+/H+ exchangers of the renal tubule]. AB - Angiotensin II is a key element in regulating the volume of extracellular liquid. It acts indirectly through aldosterone secretion by adrenals and directly on the renal tubule too: It regulates luminal Na+/H+ antiporters (NHE3 and possibly NHE2) after binding to membrane AT1 receptors located both on the basolateral and on the apical side of the cells. The main site of Ang II action is proximal tubule, mainly the S1 segment which has a high level of AT1 receptors. Circulating Ang II concentrations (10(-12) to 10(-10) M), increased NaCl, water and NaHCO3 reabsorption via NHE3 in the proximal tubule. There is also a synthesis of Ang II within the cells of proximal tubule, which is secreted within the lumen where the physiological concentration is stable 10(-8) M, i.e. 100 to 1000 times higher than the circulating concentration. Luminal ANG II originating from kidney has a physiological autocrine function on NaCl, water and probably NaHCO3 reabsorption, since inhibiting Ang II synthesis, by conversion enzyme inhibition, or effect, by AT1 receptor antagonists, induces a reduction of proximal tubule reabsorption. The stimulatory effects of circulating and intrarenal Ang II seem to be explained by protein kinase C stimulation and possibly by a reduction of cAMP production or by a stimulation of a non-receptor tyrosine kinase. When pharmacological doses of Ang II (> 10(-8) M) are applied in the peritubular or the luminal medium of isolated microperfused proximal tubule in vitro, a paradoxical inhibition of NHE3 was observed. These effects appear to involve arachidonic acid metabolites through the cytochrome P450 pathway and possibly a rise in cytosolic free Ca++. The physiological significance of these supraphysiological effects are unknown. PMID- 9857379 TI - [Active metabolites derived from angiotensin II]. AB - It has been recently shown that angiotensin II (Ang II) is not the only active peptide of the renin-angiotensin system. Several of its degradation products including Ang III (obtained by deletion of the N terminal amino acids), Ang IV (obtained by deletion of the two N terminal amino acids), and Ang II (1-7) (obtained by deletion of the C terminal amino acid), also possess biological functions. These peptides are formed via the activity of several enzymes: angiotensin--converting enzyme, aminopeptidases A and N, neutral endopeptidase and prolylendopeptidase. Ang III possesses most of the properties of Ang II and shares the same receptors AT1 and AT2. In addition this peptide is particularly important in brain physiology and plays a major role in the secretion of arginine vasopressine. Ang IV possesses its own receptors distinct from AT1 and AT2. Some of its effects (for example, stimulation of the synthesis of the type 1 inhibitor of plasminogen activator by endothelial cells) were previously attributed to Ang II. Others effects, like renal and cerebral vasodilatation, are opposed to Ang II effects. The role of Ang IV in renal physiology remains to be determined. Ang II (1-7) exhibits direct and indirect effects, the latter resulting from Ang II (1 7)-dependent formation of nitric oxide and vasodilatory prostaglandins. Ang II (1 7) potentiates the hypotensive effect of bradykinin and plays also a major role in the control of the hydroelectrolytic balance. It possesses its own receptor: AT1-7, recognizable by (sar1-thr8) Ang II or Sarthran. Finally Ang II (1-7) is converted into Ango II (1-5), by angiotensin-converting enzyme. This peptide is inactive. All of these enzymes, peptides and receptors are present in kidney. Thus the renin-angiotensin system appears to be much more complicated than thought a few years ago, setting the problem of new therapeutic tools for the treatment of hypertension and glomerulosclerosis. PMID- 9857380 TI - Role of angiotensin in the development of the kidney and urinary tract. AB - When perfusion pressure to the kidney falls, e.g., as a result of dehydration or mechanical hindrance to the renal arterial blood flow, the release of renin, hence angiotensin (Ang), surges. This feedback regulation is geared to preservation of renal hemodynamic environment by raising systemic blood pressure. We are aware that a surge of renin-angiotensin release also occurs when there is a mechanical hindrance to urine outflow. This phenomenon of ureteral pressure sensitive activation of renin-angiotensin has been heretofore viewed as an error of nature. We have obtained evidence which challenges this traditional view when we examined strains of mutant mice which are completely devoid of either angiotensin type 1 (AT1) receptor gene (Agtr1-) or angiotensin type 2 (AT2) receptor gene (Agtr2-) as a result of genetic manipulation of these animals. These strains of mice display varying degrees of urinary tract obstruction. In Agtr2- mice obstructions develop during early kidney ontogenesis in ureto, and, in Agtr1- mice, during late ontogenesis ex utero. One may recall that, throughout its normal ontogenesis, the kidney is twice at risk for obstruction of urine outflow. Thus, in utero the ureter is transiently obliterated. This transient obliteration is believed to protect the kidney from the high pressure from the cloaca when urine is not yet formed. During this period, the ureter is surrounded by dense layers of undifferentiated mesenchymal cells. Subsequent expansive growth that the ureter must achieve, therefore, in concert with a timely disappearance of the surrounding mesenchymal cells. The study in Agtr2- embryos indicated that Ang, through the Agtr2 receptor, promotes disappearance of these mesenchymal cells, and that inactivation of this receptor results in congenital obstructive nephropathy. Our additional studies in human specimens indeed indicate that many infants with congenital anomalies of the kidney and urinary tract have a significant mutation within the AT2 gene. Once animals are born, the kidney comes to be of primary importance for preservation of body fluid homeostasis, and urinary output increases dramatically. The large volume of urine predisposes the kidney to obstructive nephropathy due to the high resistance offered to the urine by the downstream ureter. Normally, a special device develops within the urinary tract in a timely fashion, which enables the kidney to collect a bulk of urine, and then to expel it downward periodically without imposing positive pressure upon the renal parenchyma. This special device is the renal pelvis. In the studies on Agtr1 null mutant mice, we learned that Ang, through the AT1 receptor, promotes development of the pelvis shortly after birth, so that inactivation of this receptor in Agtr1- mice leads to absence of development of the pelvis, hence to obstructive nephropathy. Collectively, Agtr1 or Agtr2 null mutant mice suffer from urinary tract obstruction. Given that urinary tract obstruction per se is a potent stimulus for Ang generation, Ang is essential for the kidney to escape from obstructive injury. PMID- 9857381 TI - [Interactions between the renin-angiotensin system, nitric oxide and endothelin]. AB - The renin-angiotensin system plays a major role in the regulation of blood pressure and sodium balance. Nitric oxide (NO) and endothelin (ET-1) are involved in the regulation of renin release and modulate the vasoconstrictive and fibrogenic effects of angiotensin II. the mechanisms that activate renin production are less effective when endogenous NO synthesis is inhibited. In the absence of NO, ET-1 prevents renin secretion. Angiotensin II stimulates the production of NO and ET-1 by endothelial cells. The vascular effects of angiotensin II are inhibited by NO reinforced by ET-1. The stimulation of ET-1 secretion could partly explain the long-term effects of angiotensin II on vascular remodelling. PMID- 9857382 TI - [Effects of angiotensin ii on vascular remodeling]. AB - Classically the AT1 receptors to angiotensin II are considered to be present on the smooth muscle cell membrane in the arterial wall, in which they diffusely regulate peripheral resistances. They are also present on numerous other cell types, including fibroblasts and myofibroblasts, monocytes and macrophages, endothelial cells, where they participate to the phenotypic modulation of the cell, involved in their activation leading to tissular remodeling. The intra cellular pathway involving the phospholipase C activation and the mobilization of intra-cellular calcium is predominantly involved in the functional vasomotor response to angiotensin II. In contrast the intra-cellular signaling pathway leading to production of oxygen free radicals and activation of the NF-kappa-B system is probably mainly involved in the phenotypic modulation of target cells and their consequences on the vascular tissue remodeling. PMID- 9857383 TI - Angiotensin II is involved in the progression of renal disease: importance of non hemodynamic mechanisms. AB - Several recent studies have provided clear evidence that angiotensin-converting enzyme (ACE)-inhibitors slow the progression of renal disease. These effects are mainly independent from a comitant reduction in systemic blood pressure. Thus, angiotensin II (Ang II) exerts other effects on the kidney which are involved in the loss of renal function. Ang II induces proliferation of cultured mesangial and glomerular endothelial cells. Our group was the first to demonstrate that Ang II stimulates hypertrophy of cultured proximal tubular cells. Ang II stimulates bioactivation and expression of transforming growth factor-beta (TGF-beta) in tubular MCT cells. This Ang II-mediated expression of TGF-beta is due to an increase in transcriptional activity. A neutralizing anti-TGF-beta antibody attenuates the Ang II-induced increase in protein synthesis in MCT cells suggesting that the hypertrophy is mediated by synthesis and activation of endogenous TGF-beta. Proximal tubular cells undergoing Ang II-mediated hypertrophy are arrested in the G1-phase of the cell cycle and express typical G1 phase-associated genes. Induction of such G1-phase-associated early growth response genes have been also described in vivo after infusion of Ang II into the renal artery. This G1-phase arrest depends on the induction of the cyclin dependent kinase (CdK) inhibitor p27Kip1. p27Kip1 expression is stimulated after incubation of LLC-PK1 cells with Ang II or TGF-beta and binds to cyclin D1-CdK4 complexes, inhibits their kinase activity, and hampers G1-phase exit. Ang II stimulates transcription of collagen type IV in MCT cells. In addition to the classical a1 (IV) chain, a3 (IV) collagen, which has normally a restricted localization in the kidney, is also induced. This stimulation is mediated by endogenous synthesis and autocrine action of TGF-beta because a neutralizing anti TGF-beta antibody as well as TGF-beta antisense oligonucleotides attenuate Ang II induced collagen type IV transcription and synthesis. In addition, Ang II exerts immunomodulatory effects on the kidney through the induction of chemokines such as MCP-1 and RANTES. In conclusion, Ang II has emerged as a multifunctional acting as a growth factor and a profibrogenic cytokine, and even having inflammatory properties. PMID- 9857384 TI - [Conversion enzyme inhibitors against the progression of renal and vascular diseases: yes but again]. AB - Since the discovery at the end of the seventies of angiotensin converting enzyme inhibitors (ACEI), numerous clinical data became available in the indications for which these agents were initialy developed, taking into account the putative role of renin angiotensin system: first in severe hypertension, secondly in moderate hypertension and then in heart failure. At the same time, an increasing interest was raised for "evidence based medicine" leading to a change in clinical study design: from clinical documentation of effects based on intermediate end points such as blood pressure, serum lipids, serum electrolytes or physical training capabilities to clear demonstrations through double blind placebo controlled trial with a positive effect on morbi-mortality. This evolution was furthermore stimulated by advances of knowledge on physiopathological mechanisms as well as the emergence of new drugs within this therapeutic class both stimulating clinical research. In that prospective, three examples are obvious: treatment of myocardial infarction, slowing down of the progression of diabetic nephropathy and of chronic renal failure. All these new indications for ACEI were obtained though large morbi-mortality clinical studies which are reviewed in this article. Finally, clinical studies are running with ACEI in order to demonstrate a possible effect on primary or secondary prevention of cardiovascular morbi mortality in high risk populations results which should be available at the beginning of the next century. PMID- 9857385 TI - [A plea for the "nose"]. PMID- 9857386 TI - [The nose in internal medicine]. AB - Compared with the lower respiratory tract, the nose reacts with greater physiological sensitivity to environmental stimuli by means of secretion, obstruction and sneezing. When a certain level of nasal reaction is exceeded, we have what is called vasomotoric rhinopathy. Systemic illnesses can also be reflected in the nose. Timely recognition can prevent life-threatening situations such as arise in Wegener's granulomatosis or rhinoliquorrhoea. In rhinitic complaints thought should also be given to the possible side effects of medication. Antihypertensive products and hormonal contraceptives are those most frequently responsible. The suffering experienced by a patient with chronically blocked nasal breathing is often underestimated. A variety of possible causes are reviewed. PMID- 9857387 TI - [Anagrelide in control of myeloproliferative thrombocythemia: long-term experiences in 6 patients]. AB - Symptomatic or asymptomatic chronic elevation of platelet count can be observed in all forms of myeloproliferative disorders (MPD). Benefits and limitations of the traditional platelet-reducing agents, such as radioactive phosphorus, alkylating agents, hydroxyurea and interferon alpha, are well known and have been largely described. Anagrelide (Agrelin) is an additional newer drug with a selective platelet-lowering effect. We describe our own long-term experience in 6 patients with MPD who were treated with anagrelide as part of a compassionate-use protocol between April 1991 and August 1997. The median duration of therapy was 54 months (range 17 to 75 months), with an overall response rate of 100% (complete and partial response for at least 4 weeks). The initial median platelet count of 1211 x 10(9)/l (range 847 to 2050 x 10(9)/l) was reduced rapidly and lastingly to 570 x 10(9)/l (range 216 to 667 x 10(9)/l) at the time of the last control. Under treatment with anagrelide 4 of the 6 patients showed a reduction of disease-associated symptoms or complications. Adverse reactions were generally mild and transitory, and no interruption or cessation of therapy was required. Development of drug resistance or late adverse events were not observed. Treatment with anagrelide is effective, safe and in our opinion easy to administer. It also appears to be suitable for long-term administration. PMID- 9857388 TI - [Has laparoscopy changed the surgical approach in gastroesophageal reflux? Apropos of an experience with 63 cases of gastroesophageal reflux treated by laparoscopy]. AB - INTRODUCTION: Indications for surgery in gastrooesophageal reflux disease (GERD) have decreased significantly since the introduction of proton pump inhibitors. At the same time there has been renewed surgical interest in this disease triggered by the development of laparoscopy. We present our results with laparoscopic treatment of GERD and the change in our surgical approach to the condition during the past few years. METHODS: All patients admitted to the surgical department underwent prior evaluation by a gastroenterologist. Indications for surgery were resistance to medical therapy in 30 cases, recurrence of GERD after cessation of treatment in 25 cases and personal preference of the patient when maintenance therapy was necessary in 6 cases. The data were reviewed and the patients contacted during the first trimester of 1998 to evaluate long-term results. RESULTS: 63 patients were operated on between 1991 and July 1997. Rossetti-Nissen fundoplication was performed routinely until March 1995. Thereafter, the type of antireflux surgery has been chosen according to the preoperative evaluation of the patients, including endoscopy, oesophageal manometry and gastric emptying studies. There was no mortality and, except for one case, only minor morbidity. Four patients required reoperation, two for severe gas bloat syndrome and two for recurrence of GERD. Overall, the long-term follow-up shows that 93 patients are satisfied or very satisfied with their outcome (Visick 1 or 2). 5% of the patients need daily medication for heartburn, 5% complain of occasionally disturbing dysphagia, and 12% of sometimes distressing gas bloat. More patients have gas bloat after the Rossetti-Nissen procedure than after the Nissen or Toupet fundoplications. CONCLUSIONS: The enthusiasm of surgeons for the laparoscopic approach to GERD has prompted renewed interest in the pathophysiology of the disease. Consequently, indications for surgery are better defined, surgical technique is chosen according to the individual patient based on objective preoperative studies and follow-up has improved. In this setting, laparoscopic fundoplication represents an effective treatment for GERD. It is associated with lower postoperative morbidity than open surgery and represents the first choice when surgery is indicated. PMID- 9857389 TI - [Gastroesophageal reflux in infants and children]. AB - Gastrooesophageal reflux is a common clinical condition in infancy and childhood. Evaluation and treatment are indicated if it is associated with complications such as failure to thrive, oesophagitis or pulmonary symptoms. Depending on the clinical symptoms, investigations may include pH-monitoring, upper gastrointestinal series and endoscopy. Gastrooesophageal reflux may be treated by parental reassurance, dietary advice, positional treatment, prokinetic agents and acid secretion inhibitors. Surgery is rarely indicated. PMID- 9857390 TI - [Fluctuating consciousness with diagnostic CT]. PMID- 9857391 TI - Understanding the continuum of discrete-trial traditional behavioral to social pragmatic developmental approaches in communication enhancement for young children with autism/PDD. AB - Clinicians are faced with the challenge of making informed decisions amidst heated debates over the most effective treatment approaches for young children with autism. This article provides a more specific focus to this debate by considering the practice of enhancing spontaneous language and related social communicative abilities of young children with autism/pervasive developmental disorder (PPD). First, a historical perspective of the evolution of different approaches for enhancing communication and related abilities is presented, followed by a description of characteristics of the approaches. The approaches are described along a continuum from massed discrete trial, traditional behavioral to social-pragmatic, developmental. The current state of knowledge regarding the effectiveness of early services for children with autism/PDD is examined and conclusions are presented with consideration of the need for more meaningful outcome measures than are currently used for the next generation of outcome research. PMID- 9857392 TI - Pivotal responses and the natural language teaching paradigm. AB - This article discusses the identification of pivotal behaviors for maximizing the impact of intervention for children with autism. Language deficits are not likely to make sufficient improvement unless pivotal variables, such as motivation, are addressed in the design of intervention programs. Pivotal target behaviors related to such children's motivation to engage in social communication are discussed, and their integration into a Natural Language Teaching Paradigm is described. Of particular interest are variables related to child choice, the use of natural reinforcers, the interspersal of maintenance trials to build behavioral momentum, and reinforcing communicative attempts. When used in combination to motivate the children to engage in and initiate communicative interactions, the impact on development can be considerable in terms of rate of acquisition, generalization of gains, and normalization of language development. PMID- 9857393 TI - The picture exchange communication system. AB - The Picture Exchange Communication System (PECS) was developed as a means to teach children with autism and related developmental disabilities a rapidly acquired, self-initiating, functional communication system. Its theoretical roots combine principles from applied behavior analysis and guidelines established within the field of alternative and augmentative communication. This approach has several potential advantages relative to imitation-based strategies (both vocal and gestural) and symbol selection strategies. The system begins with the exchange of simple icons but rapidly builds "sentence" structure. The system also emphasizes developing the request function prior to developing responding to simple questions and commenting. The development of requesting with a sentence structure also permits the rapid development of attributes more traditionally taught within a receptive mode. The relationship between the introduction of PECS and various other behavioral issues (i.e., social approach and behavior management) as well as its relationship to the codevelopment of speech are reviewed. PMID- 9857394 TI - Peer-mediated social intervention for young children with autism. AB - In this paper, we trace the history of peer-mediated intervention for young children with autism. Special attention is paid to outcomes achieved, implementation strategies, practicality, combining peer-mediation and naturalistic teaching, and overall conclusions from 20 years of study. PMID- 9857395 TI - Environmental supports to enhance social-communication. AB - "Environmental support" is a general term used to define the systematic use of explicit visual and/or social cues to clarify meaning for children with autism. Environmental supports can serve as a compensatory means to structure and enhance language understanding, social engagement, and social-communicative interactions. The unique and atypical developmental patterns associated with autism provide the rationale for the use of two environmental supports: visually cued instruction and modified sociolinguistic input. Using empirical research, multiple examples of environmental supports to enhance social-communication are provided. PMID- 9857396 TI - [Health promotion and public health]. PMID- 9857397 TI - [Program of action by the Swiss Foundation for Health Promotion/1998-2002]. PMID- 9857398 TI - [Pedalo--a community project for mothers of infants in St. Gallen-Rotmonten]. AB - The "Pedalo" pilot project has been in the trial phase in St. Gallen's Rotmonten Quarter for a year and a half now. It was developed by a group of women residents of the quarter. Having the backing and support of the ZEPRA executive committee, the objective of the project is to reunite mothers with one or several small children and offer them support and guidance in everyday and crisis situations. The project is designed as a grassroots effort with a preventative character, working hand in hand with trained laypersons within Rotmonten and joining forces with other support organizations and agencies of the City of St. Gallen as needed (catalyst and coordination function). The model for the "Pedalo" meeting place was the Dutch intervention project Moeders informeren moeders (MIM, "Mothers Inform Mothers"). This project idea was adopted by a group of mothers who had previously taken the initiative in their own neighborhood. Today, "Pedalo" essentially provides for a meeting place, in addition to organizing ad-hoc events or campaigns in the neighborhood. Since the beginning of the trial phase the project has been supported by public funding and private donations. PMID- 9857399 TI - [The "stop violence" campaign of the Swiss Conference of Equality Officers]. AB - The campaign "Stop Domestic Violence against Women" (referred to below as "Stop Violence") is a prevention campaign of the Swiss Conference of Equality Officers. It has been primarily focussed an heightening public awareness of violence against women in relationships with the opposite sex and offering victims and experts a refuge and meeting place. Central elements of the campaign: public service announcements on television and in the radio broadcast at various times of the day and the Info Line, a trilingual hot line and contact point specially set up by the campaign for victims and experts. The evaluation was designed to describe and estimate the type and extent of phone contacts with victims, experts and other callers. It was also designed to survey the opinion of the campaign on the part of those confronted with the victims of domestic violence on a daily basis, i.e. the staff of women's shelters. Consequently, the goal of the evaluation was to gather feedback on the campaign as well as assess the benefits and effect of the Info Line. PMID- 9857400 TI - [A quality culture for health promotion!]. AB - Quality and quality management are omnipresent as notions in the daily life of projects but there exists neither a clear conception nor a homogeneous quality understanding. This expresses the fact that a critical and self-reflecting project culture is not rooted strongly enough. A good project and quality management is indispensable to optimize the goal attainment. Up to now, however, concrete implementations as far as quality improvement in the daily lift of projects is concerned, were lacking. This article shows the necessity of health promotion being confronted with quality and what can be gained by that. It presents a project of the Institute of Social and Preventive Medicine of the University of Zurich in which quality promotion is specifically put into practice. In co-operation with project leaders, instruments for a quality management are being developed and tested which support and steer the constant process of quality promotion. PMID- 9857401 TI - [Health promotion--as a change of social systems]. AB - Most prevention concepts are principally based on the goal to change people's behavior, yet consider structural changes as important. Often measures concerning structural changes are planned and realized over people's heads. The concept of health promotion according to the Ottawa Charta postulates the planning and realization of structural changes with the participation of the persons affected by it. Methodical hints and concrete examples of such participative changes of social systems (communities, districts, schools, hospitals, firms etc.) are rare in literature. This article outlines how, with the model of the "Public-Health Action Cycle", changes in social systems can be participatively planned and realized, with the aim of developing or maintaining healthy social systems. Social systems can only be changed together with the people affected (participation), if these systems are comprehensible. In this way, participative structural changes become a practicable goal in health promotion. PMID- 9857402 TI - [Health promotion in the basic education of teachers in Switzerland]. AB - School is an extraordinary setting through which to practice health promotion. Obviously the preparation of future teachers to this task is of great importance. In Switzerland, there are some models of good practice and many enthusiastic fighters for health in the field of pre-service teacher training. Yet the results of a study at the swiss teacher training institutes show that about 30% of these institutions do not include health-issues in their curriculum. Furthermore, in those institutes including health-issues, the structural position of the person(s) encharged with this task is often weak and marginalized. The practical impact of the study is only partially satisfying: while the addressed authorities (Swiss Conference of Cantonal Ministers of Education, Cantonal school authorities) have translated and published the results of the study and give them some weight by formulating their own recommendations, the constitution of a network of the persons encharged with health-issues in the teacher training institutions has failed. PMID- 9857403 TI - [Health promotion and prevention in the Lucerne canton--1995 analysis and prospects]. AB - Considering the legal situation the canton of Lucerne is addressed to make his contribution to health promotion and prevention. His target groups are authorities on cantonal and community levels, specialised organisations and other health professionals, as for example physicians and pharmacists. The objectives of this study are to survey the health promotional and preventive activities, to evaluate the needs of the target groups as well as to focus on the opportunities of improved cooperation. Based on these results recommendations to the cantonal health authorities will be formulated. As far as the allocation of financial and personal resources are concerned health promotion and prevention do not figure as top priorities. Besides, the study results suggest to improve the cooperation between political representatives and health professionals. The cantonal health promotion authorities are well known among the target groups. However, cooperation could be intensified on all levels of action and is regarded as most desirable. A mutual forum focused on information and experience transfer would support these results and could stimulate joint projects. In addition, the needs of the communities have to be given special attention. PMID- 9857404 TI - [Health promotion in the neighborhood--resources and needs of neighborhood groups and institutions]. AB - The goal of the present study was to assess how inhabitants and organizations perceive health resources and problems in their neighbourhood, and where they see needs and possibilities to improve the situation. In two areas of the city of Berne 617 inhabitants were interviewed in the street, and 95 neighbourhood organizations answered a postal questionnaire. Most resources and problems were related to the environmental situation, infrastructures, and social life in the neighbourhood. Noteworthy findings were the high degree of insecurity mainly of older women, the lack of integration of many foreign inhabitants, and the difficulties of living together among different generations and nationalities. The inquiry among neighbourhood-organizations confirmed their importance in several health related aspects of the neighbourhood: e.g. social support, lobbying for local interests, and recreational activities. Many of these organizations can be motivated for further participation in health promotion: by increasing their sensitivity to health implications of their work, by improving collaboration and networking, and by appropriate professional support. PMID- 9857405 TI - [Evaluation of a community-oriented health promotion project "Mauren Aktiv"]. AB - In the year 1996, "Mauren Aktiv"--a community health promotion project in Liechtenstein--was evaluated. The methods used were as follows: Interviews with key-persons of the community, group-discussions, written questionnaire, telephone interviews and analysis of documents. The evaluation showed the following results: "Mauren Aktiv" works. It has increased the offer of health promotion activities by more than 100 percent. The offer is well received by the population. The flexible organisational structure proved to be functional. But "Mauren Aktiv" only partly achieves its claim to be a co-ordinating agent for all health promotion activity within the community. Favourable factors for the project are the co-operation of numerous volunteers and the conditions offered by the municipality. On the contrary the co-operation of the volunteers and the lack of professional support cause problems, for example regarding the documentation of the activities of "Mauren Aktiv". PMID- 9857406 TI - [New drugs for horses and agriculturally useful animals and changes in the pharmaceutical market since 1996]. PMID- 9857407 TI - [Case report. Increase in bulk of the inguinal area in a pig]. PMID- 9857408 TI - [Laparoscopic reposition and fixation of the left displaced abomasum in cattle]. AB - This article explains a minimally invasive technique for surgical correction of the left displaced abomasum in cattle. Endoscopic photographs show how the abomasum will be deflated, replaced and percutaneously fixed. Laparoscopy makes it possible to fix the fundus of the abomasum in all clinical conditions, independent of size, gas filling and fluid in abomasum and rumen. Especially cases with extremely dilated abomasum and cases with small quantity of gas within the abomasum are no problem in percutaneous abomasopexy. Fixation of the pyloric part of the abomasum or other abdominal structures are avoided. Adhesions of the abomasum to the rumen or the left body wall and other pathologic conditions can be seen. PMID- 9857409 TI - [Use of sodium ceftiofur in the combined therapy of complicated septic diseases in cattle]. AB - The use of Sodium-Ceftiofur (Excenel) with a dosage regimen of 1 mg/kg body mass intramuscularly was evaluated in the therapy of complicated claw diseases, like septic arthritis of the distal interphalangeal joint, septic tenosynovitis of the digital flexor tendon sheath or complicated interdigital necrosis. Sodium Ceftiofur was applied in 34 dairy cattle in addition to surgical intervention. 14 cattle of the control group perceived only surgical intervention without additional antibiotic therapy. Application of the cephalosporin proved to be necessary, with the untreated animals exhibiting far more disturbance of the reconvalescence course in the postoperative phase. Treated animals had a decreasing duration of the postoperative stay at the clinic. Sodium-Ceftiofur was found to be an effective antibiotic in the therapy, an application over a five- to 10-day-period is recommended in complicated digital diseases. As a side effect five cows exhibited slight diarrhea, which disappeared without additional treatment. PMID- 9857410 TI - [Thyroid hormones in Old German Black Pied- and Holstein-Friesian suckling calves. Effects of parturition and postpartum adaptation]. AB - The present trial deals with the effects of birth stress on the thyreoidea respectively on the thyroxine- and triiodthyronine-concentration as well as on their unbound fraction (FT4 and FT3) of suckler calves and the evolution of those parameters in the first three months of life. T4- and T3 concentrations in blood serum of calves were beyond maternal levels increasing significantly in the first 24 hours with strong individual specificity and reaching levels of adult animals at the end of the three-month-period. The different increasing rates of the four parameters show a stimulation of the thyreoideal activity and of the monodejodinase-system as well and they show also a greater decomposition rate of the binding proteins. An influence of the course of parturition on the T3 level p.n. and the T4/T3 ratio in the first 24 hours could be disclosed. The hormone concentrations p.n. showed correlations to the pH and pCO2 of the neonatal calves. For T4 and T3 a strong influence of maternal levels on the hormone levels of the calves p.n. could be ascertained. Effects of breed, surrounding temperatures and type of husbandry on the investigated parameters could be established. PMID- 9857411 TI - [Occurrence of struma in cattle in the Czech Republic]. AB - In the years 1988 to 1995 among 1355 cattle examined in the frame of iodine deficiency studies in the Czech Republic 404 animals (30%) showed congenital struma. Clinical and postmortal findings are described. A monitoring of iodine content in milk showed lower values in herds with struma prevalence. The insufficient supply of iodine in these animals results from low iodine content of feed or from the goitrogenic influence of nitrate in drinking water or high content of crucifera in feed or other stress factors. An increased iodination of mineral mix in feed is highly recommended. PMID- 9857412 TI - [Clinical examinations of gilts with puerperal septicemia and toxemia]. AB - In a clinical examination on 78 gilts suffering from feverish puerperal illness the signs were documented. 42 gilts without puerperal disturbances out of the same farms (n = 22) were used as a control. Data for statistics were recorded in minute-books, listing 38 parameters, partly divided in three or four subunits. Within the first 24 hours after parturition 60% of the probands were registered, the other gilts up to the 72nd hour after parturition. 42.3% of the patients beared for the first or second time. Duration of partus averaged more than six hours in 85.9% of patients with consequent puerperal illness, whereas probands of the same age in the control group finished parturition in less than three hours (78.8%; p < or = 0.01). Frequency of obstetrical intervention measured 27% in the group of patients, in the group of probands 9.5% (p < or = 0.05). Gilts with following puerperal illness delivered 1.1 piglets more than healthy individuals (p < or = 0.05) and showed an increased stillbirth rate (p < or = 0.01). Signs of clinical interest in puerperal illness were increase of body temperature (p < or = 0.01), cardiac rate (p < or = 0.01) and respiratory frequency (p < or = 0.01). Approximately 75% of the patients showed anorexia, 66% abnormal faecal consistency. 24.4% of the diseased animals showed exclusively signs of mastitis, in 29.5% there was a combination of mastitis and inflammatory affection of the genital system to be diagnosed, in 46.1% of the cases a solitary infection of the reproductive tract was to be stated. Predominantly E. coli, followed by Staphylococcus spp. and Streptococcus spp. were isolated from the genital tract. The results of this study emphasize the clinical necessity to differentiate between isolated mastitis and puerperal septicaemia respectively toxaemia in cases of feverish puerperal illness. Puerperal septicaemia and toxaemia can, but do not have to be associated with mastitis in gilts. PMID- 9857413 TI - [Radiologic description of the growth plates of the atlas and axis in foals]. AB - Fractures of the first two cervical vertebrae, atlas and axis, may occur in foals for different reasons, e.g. in cases of a fall, going head over heels or when being hit by a hoof. The tentative clinical diagnosis can be confirmed by x raying the standing animal, with aid of computed tomography in the anaesthetized foal respectively. The growth plates however, and their time of closure have to be considered when interpreting radiographs. In the atlas there are two ventrolateral plates and one dorsomedian cartilagineous plate. Only the dorsal plate, however, can be found in the dorsoventral projection up to an age of about 12 months. The ventrolateral growth plates which have closed at about six months of age cannot be seen in either ther dorsoventral or laterolateral projection. In transversely oriented CT-scans all of the three centres of ossification can easily be made visible in the atlas. In the axis the cartilagineous gaps between the dens axis and the cranial epiphysis as well as the cranial and caudal epiphyseal growth plate can be shown in normal x-radiographs in both planes of projection. Besides this there are growth plates between the corpus vertebrae and the arcus vertebrae in the axis which cannot be seen in either radiographic projection. They become clearly visible in transversal CT-scans and are ossified at the age of three to four months. At the end of the first year the growth plates between the dens axis and the cranial epiphysis have closed. The cranial and caudal epiphyseal plate are gone at an age of about four to five years. PMID- 9857414 TI - [Diagnostic validity of thermography of lameness in horses]. AB - Thirty-six lameness free horses and 119 horses with lameness of the distal forelimb including the carpus were evaluated through thermography. Examination was done with an infrared thermography camera "Thermovision 470" and a specially developed analyzing software program. Temperature differences between corresponding regions of the left and right forelimb were determined and scrutinized statistically between various lameness groups. By creating temperature differences between both limbs a parameter for skin temperature was found which is independent of surrounding temperature. In lameness free horses skin temperature was contralaterally symmetric and there was no significant temperature difference between left and right limb. A significant (p < 0.01) temperature difference of all regions in comparison to lameness free horses was demonstrated in diseases like navicular disease, pododermatitis and tendopathia. Horses with a diagnosis of coffin bone fracture and arthropathia showed a significant (p < 0.05) temperature difference in almost all regions compared to controls, whereas horses with laminitis and periostitis did not show a significant difference. Loss of symmetric distribution of skin temperature could be demonstrated between affected and non-affected regions. Thermography can show and quantitatively prove very well changes in skin temperature in forelimb lameness. It must be emphasized that thermography in lameness diagnosis of horses is only useful in combination with a thorough clinical examination including additional examination procedures. PMID- 9857415 TI - [Dicrocoeliasis in New World camelids]. AB - Natural infections with the lancet fluke (Dicrocoelium dendriticum) were detected in a group of seven diseased or dead/euthanized South American Camelids (five Llamas, two Alpacas) from Switzerland and Southern Germany. Clinical findings in almost all the animals revealed an acute decline of general condition followed by recumbency, decreased body temperature and a varying degree of anaemia. Concurrently, all animals showed an average to poor nutritional status. All liver enzyme activities analysed in serum biochemistry conformed to the reference values and therefore offered no diagnostic clues for this disease. Necropsy however, disclosed major alterations in the liver in the form of cirrhosis, abscesses, granulomas, and a massive infestation with D. dendriticum. The coprological investigations performed at the outset of the examinations revealed eggs of the lancet fluke in only two animals. This suggests that clinical findings alone permit at best only a provisional diagnosis. Repeated coprologic follow-ups showed that the presence of eggs of D. dendriticum can be diagnosed accurately and that clinical signs appear with an excretion rate above 1000 eggs per gram faeces (EpG). In these cases, praziquantel in a single dose of 50 mg/kg per os was given. This treatment was well tolerated and achieved a quite acceptable 90% reduction of eggs in the faeces. PMID- 9857416 TI - [Sevoflurane (SEVOrane) as an inhalation anesthetic in dogs in comparison with halothane and isoflurane]. AB - 1969 Sevofluran was synthesized and in December 1995 licensed for clinical use in Germany. The low blood/gas partition coefficient is responsible for the fast uptake and elimination of sevoflurane. Sevoflurane does not irritate the airway. In human medicine no side effect of liver- and kidney function have been seen after sevofluran anaesthesia. There is low cardiovascular and respiratory depression caused by sevoflurane. In this study the use of sevoflurane in dogs should be tested and compared with isoflurane and halothane anaesthesia. All dogs were premedicated with /-methadon and diazepam. No significant depression of the cardiovascular system was seen. Neither kidney-nor hepatotoxic side effects could be found after sevoflurane, isoflurane and halothane anaesthesia. After sevoflurane anaesthesia the dogs woke up quietly and without any excitation and were able to stand on average ten minutes earlier after sevoflurane anaesthesia than after isoflurane and 85 minutes earlier than after halothane anaesthesia. PMID- 9857417 TI - [Possibilities and status of heart surgery in dogs]. AB - Before use of cardiovascular surgical techniques and procedures in humans, many experiments, e.g., hypothermic circulatory arrest and cardiopulmonary bypass using the heart-lung machine, have been performed in the dog. As a consequence experimental canine cardiovascular surgery is highly developed. This has not resulted in the routine performance of open heart surgery in veterinary medicine, probably because of the high costs. Cardiovascular surgery in the dog is generally limited to interventions not depending on hypothermic circulatory arrest or cardiopulmonary bypass. The clinical cardiovascular surgery in dogs can be divided into routine and more specialized interventions. The first category includes correction of peritoneopericardial diaphragmatic hernia, pericardial fenestration in dogs with pericardial effusion, treatment of persistent right aortic arch, and patent ductus closure. The specialized interventions include dilation of pulmonic and aortic stenoses and pacemaker implantation. The diagnosis and surgical treatment of such diseases is described. New developments in cardiovascular surgical treatment that can be expected include catheter techniques for occlusion of shunts and dilations using balloons, because the financial costs of these procedures are not prohibitive. PMID- 9857418 TI - [Case report. Extensive discharge skin changes in a cat]. PMID- 9857419 TI - [Ophthalmologic quiz. Enophthalmos and blepharospasm in a dog]. PMID- 9857420 TI - [Disorders of sex development in dogs]. AB - In two prepubertal dogs of female phenotype, a clitoral enlargement with a penile structure leading to recurring inflammation of the vagina was diagnosed. Progesterone, oestrogen and testosterone serum concentrations were radioimmunologically analysed. The histology of the penile structure, the gonads and the uterus was made via a hematoxylin-eosin staining method. The chromosome constitution being analysed by the method of Basur and Gilman (1964) was 2n = 78,XX. Regarding all results of this investigation the diagnose was XX-male. The development of this sexual disorder is discussed. The surgical removal of gonads and uterus and the excision of the enlarged clitoris is described. PMID- 9857421 TI - [Ehrlichia canis infections of dogs in Germany. Epidemiology, diagnosis, therapy and prophylaxis]. AB - Between January and December 1997 infections with Ehrlichia canis were detected in 211 dogs in Germany. Of the 53 epidemiologically evaluable dogs, 19 animals born and raised in Germany had travelled with their owners abroad in endemic areas, 30 dogs originated from there and four dogs had never left Germany. As regards to the possible location of infection it has been registered that most dogs had been taken to countries of the Mediterranean Sea (Spain, France, Italy, Greece) or had been imported from there. On inquiry, ticks had been forwarded from four dogs only, which were determined as Rhipicephalus sanguineus. The subsequent serological investigation of these four dogs revealed a Rickettsia conorii infection in two of them. PMID- 9857423 TI - [Abnormal behavior and adaptation problems in dogs and cats and their pharmacologic control]. AB - Small animal practitioners are increasingly confronted with patients showing adaptation related problems (ARP) which are expressed as disturbed or abnormal behavior (DAB). As a result, practitioners are asked increasingly to euthanize animals which seemingly cannot be socialized. In healthy dogs and cats, three main causes for DAB can be detected: refusal of obedience because of the drive for dominance; anxiety and frustration; and geriatric DAB. Increasingly, disease conditions not readily diagnosed can cause DAB, especially hypothyroidism. Influencing and contributing factors to DAB are breed, sex, experiences as a puppy, behavior of owners, changes in the pet's environment. ARPs may also cause disturbances in the condition of skin and fur, e.g. atopic dermatitis, pruritus sine materia, lick granuloma, and of the intestinal organs (vomiting, irritated bowel syndrome) and may result in an immune deficiency. Therapeutic approaches include behavioral therapy, surgical or hormonal castration with progestins or antiandrogens, substitution with thyroxin in cases with hypothyroidism, and/or the use of psychopharmaca, most prominently of modern antidepressiva like amitriptyline; buspirone; clomipramine and fluoxetine, but also of selegiline, a mono-aminoxydase inhibitor. These compounds, among other effects, are elevating prolactin levels. This seems to allow to formulate a working hypothesis: in the canine species, prolactin is obviously a hormone enabling socialization; hence all drugs which safely cause an increase in prolactin production might be suitable to manage or control ARPs and DAB in the dog, but also in the cat. Higher levels of prolactin than those required for socialization, as seen in nursing bitches or some clinically overt cases of pseudopregnancy, may cause maternal aggression and can be controlled with prolactin inhibitors, if needed. PMID- 9857422 TI - [Extensive stomach ulcers due to Dieffenbachia intoxication in a cat]. AB - Extensive gastric ulceration was diagnosed endoscopically and histologically in a two-year-old cat with haematemesis, anorexia and severe dehydration. Based on history (previous ingestion of leaf pieces of Dieffenbachia) and exclusion of another cause, intoxication with Dieffenbachia was diagnosed. The toxical effect of Dieffenbachia on mucous membranes is probably explained by ejection of calcium oxalate crystals leading to injury of mast cells and a massive histamine release. Supportive treatment including electrolyte infusions, ranitidine, sucralfate, and forced feeding using an endoscopically placed gastrostomy tube resulted in quick clinical recovery of this cat. PMID- 9857424 TI - [Cystic testicular anomaly with subsequent stenosis of the small intestine in an emerald tree boa (Corallus caninus L., 1758)]. AB - A case report with emphasis on diagnostic procedures and differential diagnoses is given. The etiology is discussed in comparison to similar malformations in mammals and other reptiles. The literature is reviewed with emphasis on differential diagnostic aspects. PMID- 9857425 TI - [Oleothorax: the "bomb" from the past is still ticking]. PMID- 9857426 TI - [Prognosis for cardiopulmonary resuscitation]. PMID- 9857427 TI - [Advances in the treatment of chronic lymphocytic leukemia (CLL)]. AB - CLL is a chronic lymphoproliferative disorder which is characterized by the proliferation of a CD5 positive B cell clone. At diagnosis most patients are in early stage of the disease (stage A). It is well established that an early treatment in stage A is not associated with a survival advantage. Therefore, the disease should be treated only when signs of progression are present. The standard initial therapy is chlorambucil +/- prednisone. By this treatment remissions (mostly partial remissions) are achieved in about half of the patients. Fludarabine, a purine analogue, is more effective than chlorambucil as initial treatment (75% remissions, 27% complete remissions, 33 months progression free survival). However, the overall survival is not prolonged when compared to chlorambucil treatment. Fludarabine has a high efficacy in patients with recurrent disease, in particular in those who are resistant to alkylating agents. The allogeneic and autologous stem cell transplantation is an experimental treatment which may be indicated in younger patients who show a response to conventional treatment. With both procedures hematological remissions are frequently achieved and in some of the patients long lasting molecular remissions were obtained. Currently, it is unknown whether patients can be cured by stem cell transplantation and longer follow-up will be necessary to clarify this question. Monoclonal B-cell antibodies are effective in vivo and partial remissions can be obtained in chemotherapy refractory patients. Antibodies have a high efficacy in clearing lymphoma cells from peripheral blood and bone marrow but are less effective on the organomegaly. PMID- 9857428 TI - [Limitations of life support in patients with anoxic encephalopathy after CPR. Results of a questionnaire to Austrian intensive care units]. AB - A questionnaire was sent to 97 Austrian intensive care units to define the current practises of predicting neurologic outcome in patients with anoxic encephalopathy after cardio-pulmonary resuscitation. All relevant prognostic predictors, such as clinical scales, electrophysiological techniques and laboratory tests were included to examine if these procedures influence the decision to withdraw life support. The answers of 68 (70%) units were collected and analysed. The results show that tests such as somatosensory evoked potential of the determination of brain cell-specific cytosolic enzymes are without any practical importance in Austria. In addition, the methods of reducing life support were evaluated. According to the controversial discussion in the literature, there is no common approach to this problem, especially in the question of withdrawing controlled mechanical ventilation. The result of our survey show, that there is a need for general guidelines to determine the prognosis and the methods of withdrawing life support which could be used in all intensive care units regardless of their size and technical infrastructure. PMID- 9857429 TI - [Imipenem resistance in Pseudomonas aeruginosa]. AB - In 1997 in western Austria, 9.9% of Pseudomonas aeruginosa strains from patients of general practitioners were resistant to imipenem as well as 18.2% of the isolates from hospitals and 20.2% of the strains at a university teaching hospital. Within the hospital the imipenem resistance varied from 9.9% among out patients to 28.7% in isolates from intensive care units. In medical/surgical words, up to 15.1% of P. aeruginosa strains were resistant to imipenem. The incidence of imipenem-resistant P. aeruginosa strains correlates to the use of carbapenems. In June 1997, 10 consecutive isolates from 8 patients were obtained and typed using restriction fragment length polymorphism analysis (RFLP) and Pyocin typing. All 10 isolates were resistant to meropenem as well as to imipenem. The finding (by RFLP and Pyocin typing) of individual bacterial types in each isolate strongly contradicts the spread of infection by cross infection. However, all patients were proven to have been treated with imipenem during the 3 months prior to testing. In 1997, 13,880 g of imipenem were used at the university hospital in Innsbruck. The use of carbapenems appears to be the main cause for the increased incidence of imipenem-resistant P. aeruginosa strains. PMID- 9857430 TI - [Management of severe infection complicating aortofemoral grafts]. AB - OBJECTIVE: Infection of the retroperitoneum after implantation of an abdomino femoral aortic graft remains one of the main problems in vascular surgery. On basis of a critical review of own experiences we evaluated the management of this difficult clinical situation. PATIENTS: From 1970-1996 1500 aortofemoral graft operations (aneurysmal disease: 512, aortoiliacal occlusive disease: 988) were performed. Abdominal infection occurred in 12 patients (0.8%) (12 men, median age 60.5 [48-80] years). RESULTS: The median interval between operation and infection was 17.7 (0.5-108) months. The port of infection was in 50% the groin, 25% suffered from abdominal infection, in 3 cases it was not to identify. Clinical manifestation of infection was aortoduodenal fistula in 2 patients, false aneurysms in 2 cases, and a paraprosthetic abscess in another 4 patients. Operative therapy comprised (partial) removal of infected material in 10 patients with consecutive extraanatomical reconstruction in 8 of these. Mortality of graft infection was 50%. Causes of death were untreatable sepsis in 4 patients, another 2 died from hemorrhagic shock. 3 out of 6 surviving patients finally lost their limbs following multiple vascular procedures. CONCLUSION: Adequate surgical therapy of infected aortofemoral grafts remains an unsolved problem. Lack of knowledge of suitable parameters for the best treatment leaves the outcome of prosthetic infection unpredictable. Removal of the infected graft with extraanatomic reconstruction seems to be the standard of surgical treatment, which is recommended in these cases. PMID- 9857431 TI - Rupture of therapeutic oleothorax leading to paraffin oil aspiration and dissemination of tuberculosis--a fatal late complication of tuberculosis therapy in the 1940s. AB - In the 1940s, oleothorax (paraffin oil instillation) was widely used to treat patients with apical tuberculosis. The oil plombage should have been removed after a few years; however, since oleothoraces were usually asymptomatic, removal was uncommon. These in the meantime elderly patients are at risk of late complications, such as rupture of the oleothorax and aspiration of oil. We report the case of a 69-year-old man with a spontaneous rupture of an oleothorax leading to oil aspiration, lipid pneumonia and culture-proven disseminated tuberculosis with fatal outcome. Unexpected positive PCR for M. tuberculosis-DNA in tracheal secretions was one of the leading signs in this case. Thus oil plombage in patients with oleothorax may be "time bombs". Primary physicians should be aware of this life-threatening complication. PMID- 9857433 TI - ["Integrated psychotherapy": a new clinical challenge]. AB - Integrated psychotherapy according to Barolin wants to be embedded in the general health system and regards the whole "bio-psychosocial entity of man" using psychical means, which can take psychotropic as well as somatotropic effect. It should come out of an isolated ivory tower and seek coordination with each and everyone dealing with the "human entity": medical (like medication, physiotherapy, etc.), nursing, social work, administration, etc. Psychotherapeutic "schools" should serve as a solid base of learning. Practical use however should go beyond schools: patient-centered instead of school centered. Integrated psychotherapy should reach all persons who need it, within their somatogenic and psychogenic problems, especially the physical ill, up to accompaniment of the dying, especially in rehabilitation, including social environment, old-age groups. For this it needs social integration accepting economic responsibility (in the sense of "social ethics") plus evaluation. Hence, theory and practice of integrated psychotherapy find themselves linked to a higher human-ethical postulate, with a marked communicative component. PMID- 9857432 TI - Fatal complication of paraffin plombage after half a century. AB - Complications following thoracic plombage for treatment of tuberculosis can be observed more than 50 years after placement of the filling. The management of these late complications is challenging and frequently requires surgical intervention. We report a patient who received a plombage in 1947. She was admitted to hospital with subfebrile temperature and hoarseness. A computed tomography scan of the chest revealed transthoracic penetration of the paraffin plombage with intrusion into the overlying soft tissue. The patient underwent excision and debridement of the paraffin wax mass followed by thoracoplasty. She then developed septicaemia and died due to multiple organ failure 23 days after the surgical intervention. Early ablation of plombage should be considered in order to prevent late complications. PMID- 9857434 TI - [Basic principles of school-overlapping psychotherapy]. AB - There are general factors (non-specific and communicative) that are used in the different psychotherapeutical schools, in different degrees of importance. We strictly differentiate between schools and methods. Plurality of methods is indispensable. However, plurality of schools seems to be of no urgent importance. PMID- 9857435 TI - [Teaching, graduate education and training in psychotherapy]. AB - Basic skills in the field of psychosocial-psychotherapeutic medicine are necessary for medical students and residents. Psychotherapy as a special discipline goes far beyond this knowledge. The most important guidelines for training in Austria, BRD and Switzerland are presented. PMID- 9857436 TI - [Integrated use of psychotherapeutic treatment methods in therapy of alcoholism]. AB - The treatment of alcoholism is more promising than commonly assumed. Its success is based on the acceptance of a long-term treatment concept over a period of approximately 2 years, the willingness to differentiate between the individual treatment courses according to their underlying individual psychopathologies as well as adapting treatment measures to the actual phases during restitution. Many years of experience with various psychotherapeutic methods have proven that not so much one certain method, but their integrative application depending on the individual situation is relevant to treatment success. Thus, during treatment, a change between supportive, confrontative, systemic and family therapy-oriented elements can occur. PMID- 9857437 TI - [Integration of drug therapy and psychotherapy concepts in treatment of depression]. AB - The treatment of depressive disorders comprises an adequate pharmacological as well as psychotherapeutical intervention. The following article will demonstrate pragmatic treatment steps and corresponding problems in relation to the time context of the disorder as well as the treatment (1. Initial contact 2. First control, 3. Intermediate treatment phase. 4. Treatment termination). Without introducing innovative treatment techniques, basic biological assumptions as well as psychological aspects of depressive disorders are combined in a pragmatic way. Furthermore, new developments in psychopharmacology are taken into regard. PMID- 9857438 TI - [Psychotherapy consultation service in a trauma hospital]. AB - Psychotherapeutic consultation work with victims of accidents is an integrating approach, which is aiming at the emotional care of injured people, who already had mental disturbances before accident and/or following the trauma. During 12 years as psychotherapeutic consultant at an Accident Hospital in Vienna the necessary interventions during traumatologic treatment of stationary patients are demonstrated. PMID- 9857439 TI - [Correlation between inpatient and ambulatory psychotherapy from the viewpoint of the family physician]. AB - Very often the position of the family doctor is being reduced to a mere "doorkeeper" (a person who just lets the patients in and out). He should, however, be the first person being addressed also by patients with psychic and psychosomatic problems, which requires a fundamental education in psychosomatic or psychotherapeutic medicine. Compared to specialists or doctors working in hospitals the advantage of a family doctor is that the patients already trust him, and, furthermore, he is familiar with the family background. He functions as interpreter of the patients' complaints, and--given a certain indication--he combines medication and psychotherapy, which are the two essential parts of psychotherapy being carried out by a doctor. If the symptoms exceed his competence, he must send the patient to a specialist who can cope with his problems. His personal advice is essential in such a case. After having been released from stationary medical treatment, the patient should have the possibility to consult his family doctor again for the sake of continuous medical treatment. A successful combination of stationary and out-patient psychotherapy can be negatively influenced by the following facts: devaluation of the family doctor's competence by specialists or offending of the family doctor's feelings of self-reliance because of a supposed failure. PMID- 9857440 TI - [Problems with sexuality, erotic aspects and love in elderly patients in medical consultation]. AB - Sexual problems, especially in the elderly, are to handle as a part of the whole personality. After an informative talk about the knowledge in the field of sexuality, we have to differ between subjective and objective problems of the patient and to find out possibilities to solve real problems and conflicts. This discussion of the problems is in every case the first step to any form of therapy. PMID- 9857441 TI - [Judges are only human as well ...]. PMID- 9857442 TI - [How early and how specifically can hypoxic-ischemic brain lesions be detected with diagnostic imaging?]. AB - Pathogenic events affecting the developing brain cause malformations or lesions, the pattern of which depend on the stage of brain development. While in the past diagnosis of these patterns was made by post mortem examinations, today advances of brain imaging allow this already during life time. The patterns of hypoxic ischemic brain injuries on magnetic resonance imaging (MRI) are well known for the older child (after progress of myelination). This paper addresses the question how early and how specific these patterns can be recognized by two imaging methods, e.g. cranial ultrasound and magnetic resonance imaging. It concludes, that neonatal MRI but also neonatal ultrasound can reliably detect major lesions but may fail in the detection of less extensive patterns. Most authors therefore conclude, that a routine use of MRI for the detection of hypoxic-ischemic lesions during the neonatal period is not recommended and should rather be reserved for later controls. PMID- 9857443 TI - [Fetal acidemia and neonatal encephalopathy]. AB - OBJECTIVE: The purpose of this study was to determine the significance of fetal acidemia for newborn encephalopathy (NEP) and for the combination of NEP and multiorgan system dysfunction (MOS). In particular the influence of acidemia will be contrasted with criteria of pregnancy and delivery. STUDY DESIGN: 248 infants delivered with cord umbilical arterial pH < 7.15 (UApH) were retrospectively studied. Infants with similar or identical neonatal characteristics formed different groups: unaffected neonatal development, NEP, NEMO (NEP combined with MOS), other diseases, infection, exitus letalis. Statistics and calculations were done by means of factor analysis, univariate analysis, Student-Newman-Keuls-test, and Chi 2-test (p < 0.05). RESULTS: Twenty seven infants with UApH < 7.15 suffered from NEP. NEP in combination with MOS occurred in 11 cases. There was no relationship between the degree of acidemia and a single NEP. Infants with NEMO differ from all other groups in their mean UApH (p < 0.05). The Apgar scores 1 min and 5 min (A1, A5) separated newborns with unaffected neonatal development from all other groups (p < 0.05). UApH-differences in the group: NEMO resulted from a combination of acute intranatal fetal distress and operative delivery and a combination of A1, A5, preterm delivery, and pathologic CTG (p < 0.01). CONCLUSIONS: The degree of acidemia impacts the occurrence of NEMO. However, UApH alone does not predict this combination of characteristics. The Apgar score should be taken into account to evaluate an acidemic UApH. In case of acidemia complex factors of pregnancy and delivery are associated with an increased risk of NEMO. PMID- 9857444 TI - [Effect of continuous intravenous tocolysis with beta 2-mimetics and magnesium sulfate on erythropoietin level]. AB - 14 pregnant women receiving continuous intravenous tocolysis for threatened miscarriage were studied for an effect of beta 2-mimetics on erythropoiesis in humans. After tocolytic therapy for 48 h, the hematokrit fell by 14%; conversely erythropoietin levels rose by 88.7%. Increased erythropoiesis was reflected in increased reticulocyte (and reticulocyte subpopulation) counts. There was a significant correlation between cumulative tocolytic dose and erythropoietin elevation at 48 h. The hypervolemia evidenced by the decreased hematocrit was due to the increased oral fluid intake and fluid retention, and was the presumed main cause of the elevated erythropoietin levels. This effect could enhance understanding of the control of erythropoietin secretion. A similar mechanism may account for the increase in erythropoietin levels in pregnancy. PMID- 9857445 TI - [Detection of abnormalities in Magdeburg according to the Mainz model]. AB - Between 09/01/92 and 08/31/95 the Mainz model to register birth defects had been tested in the city of Magdeburg at the same time when the regional register instituted in 1980 was continued. The aim of this hospital-based model for the thorough registry of malformations in neonates is to examine all newborns born in hospitals using clinical and sonographical methods by experienced physicians according to a standardized scheme. Additional medical and social data were recorded. Data on live-and stillborn as well as spontaneous abortions older than the 16th gestational week and induced abortions following prenatal diagnosis were included. During the survey 5255 neonates were examined. The calculated percentage of major birth defects (by definition) was 7.8% and therefore higher than reported before. The experience with the Mainz model in the city of Magdeburg demonstrates, that this model is basically applicable to other centres. It is possible to obtain valid data on malformation frequencies in a defined population. Special attention has to be paid to certain conditions in the composition of hospital populations. PMID- 9857446 TI - [Selenium status and bronchopulmonary dysplasia in premature infants <1,500 g]. AB - Selenium is an essential component of the antioxidant enzyme glutathione peroxidase that protects tissues against oxidative injury by detoxifying peroxides. In preterm infants the risk for selenium deficiency is increased due to insufficient selenium uptake. Low selenium uptake and as a consequence decreased glutathione peroxidase activity may result in an elevated risk for the development of bronchopulmonary dysplasia (BPD). The aim of this prospective study was to investigate the relationship between the selenium status of preterm infants < 1500 g and the incidence of BPD. We determined the selenium plasma levels by means of atomic absorption spectrometry in 34 VLBW infants (mean birth weight 1075 +/- 249 g; mean gestational age 28.6 +/- 2.5 weeks) within the first 5 days of life and later in the age of 4 weeks. The infants received mainly parenteral nutrition and were not specifically supplied with selenium. Postnatally, the selenium plasma level was 34.2 micrograms/l (17.3/50) [median (25/75% quantil)] and dropped after 4 weeks to a median value of 16.1 micrograms/l (5.2/38.4) (p < 0.001). In the infants with BPD (n = 12) the selenium concentration within the first week of life was 45.0 micrograms/l (31.5/55.6) versus 33.2 micrograms/l (20.2/42.4) in the infants without BPD. In the age of 4 weeks of life the median selenium level was not significantly different between the infants with and without BPD - 17.2 micrograms/l (10.3/22.5) versus 14.8 micrograms/l (8.8/22.6). PMID- 9857447 TI - [Axillary versus rectal temperature measurement in premature and newborn infants]. AB - We performed a prospective study to see whether axillary temperature accurately reflects rectal temperature in premature and sick mature newborns nursed in incubators and whether axillary temperature measurement is better tolerated than rectal measurement. 561 pairs of rectal and axillary temperatures were obtained in 20 infants. Mean axillary temperatures were 0.16 degree C (SD 0.33 degree C) lower than rectal temperatures. In infants < 1000 g axillary temperatures were only 0.06 degree C (SD 0.30 degree C) lower than rectal temperatures whereas in infants > 2500 g axillary temperatures were 0.37 degree C (SD 0.38 degree C) lower than rectal temperatures. Our results show that axillary and rectal temperatures are interchangeable in extremely premature infants. In newborn infants however 0.4 degree C should be added to axillary temperatures to make them comparable to rectal temperatures. Assessment of infant behaviour during temperature measurement by the nursing staff showed that infants tolerated rectal better than axillary temperature measurements. The decision which kind of measurement will be performed can be taken individually for every infant. PMID- 9857448 TI - [Intact pregnancy with concomitant hydatidiform mole of a coexisting placenta]. AB - The simultaneous occurrence of an intact pregnancy with normal placenta morphology and sonographic signs of a hydatidiform mole of a second placenta, which was diagnosed in the 14th gestational week, is described. Prenatal diagnostic and perinatologic management will be reported. The difference of the placentas will be shown by histopathology. PMID- 9857449 TI - [Kinetic therapy of severe meconium aspiration syndrome]. AB - Recent studies showed the possible positive effect of body position changes during mechanical ventilation of severe lung diseases in adult patients. In neonatology kinetic therapy is still rarely used, therefore we present this case report. A full term newborn suffering from severe MAS and peripartal asphyxia was transferred to our NICU to perform extracorporeal lung support, if necessary. After application of a natural porcine surfactant and start of inhalative nitric oxide therapy (10 ppm) a clinical stabilisation was possible. Because of hypercapnia, high frequency oscillation ventilation was introduced later on. The PaCO2 values decreased quickly. A few days later severe pulmonary secretion problems occurred, which led to atelectasis and barotrauma due to local hyperinflation. After several different ventilation strategies had failed to improve the situation, kinetic therapy in combination with conventional mechanical ventilation was started. Under this therapy-concept it was possible to reventilate the atelectatic lung areas, and quickly an improvement of oxygenation was seen. Weaning from the respirator was possible within one week. In conclusion, we think that an important progress in therapy was due to kinetic therapy. PMID- 9857450 TI - [Lipoma of the corpus callosum in a newborn infant: misinterpretation of cranial ultrasound by coincidence with thrombocytopenia]. AB - Among the very rare intracerebral lipomas, those of the corpus callosum are the most frequent. Due to the advanced technology and the frequent use of ultrasonography these lesions are diagnosed more and more often. A female neonate was admitted to our hospital because of a progressive thrombocytopenia. Pregnancy was complicated by an autoimmune thrombocytopenia of the mother. While there were no remarkable findings on clinical presentation, a sonogram of the brain revealed an area of increased echogenicity in the midline which was interpreted as an intracerebral hemorrhage. In absence of any respective clinical signs a magnet resonance-tomography of the brain was performed leading to the hypothesis of a lipoma of the corpus callosum (LCC) that could be verified by a densitometry in a cranial computer tomography (CT). Obviously, the initially performed sonogram was misinterpreted as an intracerebral hemorrhage due to the coincidence with the thrombocytopenia. At last the discrepancy of clinical and ultrasonographical findings led to the diagnosis by magnet-resonance-tomography and CT scan. Knowledge of the typical sonographic appearance of an LCC may be helpful for the differential diagnosis of this rare lesion even in fetal ultrasound. PMID- 9857451 TI - [Viability ex utero in correlation with late abortion. Objections to the position on pregnancy termination after prenatal diagnosis--outline in Frauenarzt 2, 1989]. PMID- 9857452 TI - [Fetal tachycardia with silent CTC registration]. PMID- 9857453 TI - [Genetic factors in the development of atherosclerosis]. AB - Atherosclerosis is a complex multifactorial disease of the arterial wall, dependent on genetic disposition and multiple other risk factors. There are probably several candidate genes, that determine the individual susceptibility of the vessel wall to develop atherosclerosis. In recent years, a growing number of gene polymorphisms, associated with an elevated risk of myocardial infarction, has been identified. These genes and gene clusters play a crucial role in lipid metabolism, regulation of blood pressure and clotting. In contrast to rare monogenetic diseases with severe clinical signs and symptoms (e.g. familial hypercholesterolemia), genetic polymorphisms are relatively frequent. Due to their frequency, there is a high probability that one individual carries several alleles predisposing to coronary heart disease. Genetic polymorphisms become clinically important by interacting with lifestyle, environmental factors or endogenous metabolic disorders. We have recently established an animal model of rabbits, which may prove useful in the search for new genes predisposing to or protecting from atherosclerosis. Rabbits with high atherosclerotic response (HAR) show more than 70% atherosclerotic involvement of the aorta when fed a high cholesterol diet. In contrast, rabbits with low atherosclerotic response (LAR) show less than 20% atherosclerosis in spite of comparably high plasma cholesterol levels. Preliminary studies indicate that macrophages of LAR rabbits have a high scavenger receptor activity and high apolipoprotein E expression and thus appear to be very efficient in uptake and elimination of modified lipoproteins. This may result in a more efficient removal of cholesterol from the arterial wall and thus protect the animals from developing atherosclerosis. Today we are only at the beginning of understanding the complexity of gene interaction in atherosclerosis. Further identification of genetic factors of atherosclerosis will no doubt lead to a more efficient and economic prevention of coronary heart disease in the future. PMID- 9857454 TI - [Atherosclerosis of the aorta as a source of arterial embolisms]. AB - Aortic atherosclerosis has early been recognized as a potential source of embolism. The histological finding of cholesterol clefts in small end-arteries characterized the entity of cholesterol embolism. The clinical picture was extremely variable and the diagnosis was frequently established post-mortem or by means of invasive although insensitive procedures including biopsy and angiography. Therefore, cholesterol embolism was thought to be rare. With the routine use of transesophageal echocardiography for the diagnostic workup of arterial embolism, aortic atherosclerosis was shown to be the source of otherwise unexplainable embolism. Cross-sectional studies demonstrated an independent association between prominent plaques of more than 4 to 5 mm of thickness or plaques with mobile components in the aortic arch. In follow-up studies, the risk of embolic events in patients with this kind of lesions exceeded 10% per patient year. The results of pathological studies were consistent with these findings showing that ulcerated complex plaques carry an independent risk for embolic events. Apart from spontaneous embolism, atherosclerosis of the proximal aorta was shown to be a cause of embolic complications during cardiac surgery and catheterization procedures which involve the aorta. Medical treatment for the prevention of embolism in atherosclerotic disease of the aorta has not been studied systematically. In a variant form of aortic atherosclerosis consisting of mobile pedunculated thrombi inserting on relatively small plaques, anticoagulant therapy has proved to be useful in small numbers of patients. Recurrent embolic events could be prevented and regression of the thrombotic masses has been observed. PMID- 9857455 TI - [Intramural hemorrhage of the thoracic aorta: diagnosis, therapy and prognosis of 209 in vivo diagnosed cases]. AB - Aortic dissection with no entry or false lumen flow was recently identified as intramural hemorrhage of the aortic wall (IMH). Analysis of the literature revealed 209 cases of in vivo diagnosed IMH reflecting 17% of all dissections, whereas in postmortem studies this condition is found in 4-13%. Transesophageal echocardiography, computed tomography, magnetic resonance imaging and aortography (to rule out dissection) have been applied for diagnosing IMH in 57, 49, 43 and 38% of the cases, respectively. However, diagnostic accuracy of each modality is not available to date. In 34%, IMH involves the ascending aorta (type A). The average age of patients with IMH ranges between 55 and 65 years; 65% are males. In 12%, IMH was associated with abdominal aortic aneurysm. With 87%, arterial hypertension is the predominant risk factor for IMH irrespective of its location. As suggested by the term "mediastinal apoplexy" IMH may--similar to cerebral apoplexy--result from hypertensive rupture of the vasa vasorum in the aortic media. With 97%, the majority of patients present with acute chest or back pain similar to classic dissection. Mediastinal widening is found in 83%; signs of hemothorax/pleural effusion are present in 38%, acute aortic regurgitation in 26%, pericardial effusion in 23%, acute neurologic deficits in 12%, and pulse deficits in 5%. In 18%, IMH progresses to dissection and in another 15% to rupture. In 25% and 28%, respectively, dissection and rupture occur in the ascending aorta and in 12% and 9%, respectively, in the descending thoracic aorta. The 30-day mortality of IMH is 24% (36% with type A and 12% with type B IMH; p < 0.05). With surgical repair, mortality of type A IMH is lowered to 18% compared to 60% with medical treatment (p < 0.01). In contrast, with 8% mortality associated with medical treatment, prognosis of type B IMH is more favorable without surgical intervention, the latter associated with a 30-day mortality of 33% (p < 0.05). Thus, IMH is a potential precursor of dissection and should be managed like dissection with undelayed surgical intervention in patients with type A IMH and with medical treatment in type B IMH. PMID- 9857456 TI - [Perspectives of patients with terminal renal failure: quality of life and psychological adjustment before and in the first year after heart transplantation]. AB - This prospective study was designed to compare quality of life, life satisfaction, and subjective ratings of health before and at variable time intervals after heart transplantation (HTx). 175 patients were included between February 1994 and December 1997. Every six months before and 1 1/2, 3, 6, and 12 months after HTx, they received the following standardized and validated questionnaires: German SF 36, heart failure and specific transplant symptom list, global quality of life assessment, Munich life quality dimension list, expected/experienced life changes after HTx. Inclusion criteria were the acceptance of the patient on the waiting list for HTx, good command of the German language, and a minimum age of 18 years. During data evaluation, median (Me), mean (M), and standard deviation (SD) were created from individual parameters. Quality of life was rated as very poor by 84% of patients with congestive heart failure. Only 6 weeks after HTx, 74% rated their quality of life as significantly better. Before HTx 80% were very unsatisfied about their health status and 87% about physical performance. Six weeks after HTx, this parameter improved significantly and 76% were very satisfied about their health status and 50% about physical performance. While somatic changes expected before HTx corresponded well with experienced ones, psychological improvements were smaller than expected, but one year after HTx they were significant (before: M = 3.66; SD = 0.9; Range (R) = 3.78 vs 12 months postop: M = 4.61; SD = 0.6; R = 2.67; p < 0.05). Quality of life correlated before HTx best with subjectively rated health (r = 0.61, p < 0.01) and 6 months after with satisfaction about health status (r = 0.76, p < 0.01). Only in 25% were expected improvements fulfilled regarding sexual activity/satisfaction, professional situation, and recreational activities. 90% of post-transplant patients reported physical complaints, most by effects of immunsuppression, but were coping well. The study shows that already 6 weeks after successful HTx all quality of life parameters improved significantly. Despite some unfulfilled expectations and complaints, the postoperative life situation of HTx patients appeared significantly improved. PMID- 9857457 TI - [Value of spiro-ergometry and stress echocardiography on optimized programming of the upper frequency limit of dual chamber cardiac pacemakers]. AB - Patients with a dual chamber pacemaker often suffer from chronic heart failure. The influence of the upper tracking rate on cardiac performance in patients with and without congestive heart failure is unknown. Therefore, twenty patients with a dual chamber pacemaker implanted for high degree AV block were randomly programmed to upper tracking rates of 110, 120, and 130 bpm. Oxygen uptake (VO2), work capacity, and heart rate were determined at the anaerobic threshold (AT) and at peak exercise using spiroergometry every 4 weeks. Nine patients (71 +/- 12 years) had evidence for advanced heart failure (Weber C/D, group I); 11 patients (60 +/- 6 years) had no or only mild heart failure (Weber A/B, group II). Patients in group II achieved a higher mean VO2-AT at 130 bpm (17.3 +/- 3.9 ml/min/kg) than at 110 bpm (13.7 +/- 4.0 ml/min/kg; p = 0.001). Maximum oxygen uptake and work capacity at the anaerobic threshold were also significantly higher in group II with an upper tracking rate of 130 bpm than at 110 bpm (20.5 +/- 4.5 ml/min/kg vs. 18.2 +/- 5.3 ml/min/kg, p = 0.031, and 98 +/- 29 W vs. 86 +/- 27 W, p = 0.04). In group I, mean oxygen uptake at the anaerobic threshold (VO2-AT) was higher at 110 bpm (11.1 +/- 4.0 ml/min/kg) than at 130 bpm, although of only borderline statistical significance (9.2 +/- 2.6 ml/min/kg; p = 0.052). In group I a higher upper rate decreased VO2-AT by 24%, whereas in group II the higher upper rate improved aerobic capacity by 26%. Stressechocardiography (VVI pacing with a maximum tracking rate of 130 bpm) revealed new wall motion abnormalities in all patients of group I, but only in one patient of group II. Wall motion score index increased from 1.20 +/- 0.24 at rest to 1.54 +/- 0.28 under stress (p < 0.001) in group I, but not in group II (1.00 +/- 0 vs. 1.06 +/- 0.19; p = n.s.). Thus, carriers of dual-chamber pacemakers with no or mild heart failure (Weber A/B) benefit from higher programmed upper rates. In contrast, patients with more advanced heart failure (Weber C/D) improve aerobic capacity with lower programmed upper rates. This may be caused by exercise-induced ischemia in group I as indicated by stressechocardiography. PMID- 9857458 TI - [Clinical significance of sleep apnea disorders after implantation of a cardioverter-defibrillator in patients with cardiac disease and sustained ventricular tachyarrhythmia]. AB - The purpose of our study was to determine the prevalence of sleep related breathing disorders (SRBD) in patients with an implantable cardioverter defibrillator (ICD) and to evaluate prospectively the possible influence of SRBD on arrhythmia recurrence and circadian arrhythmia variation as well as on cardiac mortality during long-term follow-up. Forty consecutive ICD recipients with cardiac disease and a documented history of spontaneous, life-threatening, ventricular tachyarrhythmias underwent full-night polysomnography and were followed for 2 years. In 16 of 40 patients (40%), SRBD were diagnosed (Apnea/Hypopnea Index (AHI) > 10); in 9 of these 16 patients (56%) central sleep apneas (CSA) occurred (in 8 of these 9 patients in combination with Cheyne-Stokes respiration). Seven of the 16 patients with SRBD (44%) revealed obstructive sleep apneas (OSA). AHI was 32 +/- 15 (12-60) in patients with CSA and 32 +/- 27 (11 86) in patients with OSA. Patients with and without SRBD were comparable concerning left ventricular ejection fraction, NYHA classification, cardiac disease, ICD indication, and concomitant medication. ICD registered ventricular tachyarrhythmias occurred in 10 of 24 patients (42%) without SRBD, in 4 of 9 patients (44%) with CSA, and in 3 of 7 patients (44%) with OSA. The numbers and circadian variation of episodes registered during follow-up in patients without SRBD, with OSA or CSA were comparable (14 +/- 25, median 4 vs 15 +/- 15, median 7 vs 4 +/- 5, median 2.5). The 2-year cardiac mortality was highest in patients with CSA (4/9 (44%) vs. 0/7 patients (0%) with OSA vs 3/24 patients (12.5%) without SRBD. Thus, the prevalence of SRBD in patients with chronic heart failure and a history of malignant ventricular tachyarrhythmias is high (40%) and the occurrence of CSA seems to be predictive for cardiac mortality in these patients. An influence of moderate SRBD on arrhythmia recurrence and circadian variation of spontaneous sustained tachyarrhythmic events could not be demonstrated. PMID- 9857459 TI - [Interventional therapy for hemangioendothelioma of the liver in a newborn infant using a central venous approach]. AB - We report on a newborn (birth weight 3600 g) with high-output cardiac failure due to a large hepatic hemangioendothelioma requiring mechanical ventilation. One day post-partum therapy with steroids, furosemide, and digitalis was initiated. Despite this, the clinical condition deteriorated. Therefore, selective coil embolization of the arterial vessels supplying the hemangioendothelioma was performed using a venous approach with passage to the arterial side via the ductus arteriosus. After the embolization the patient improved quickly and he was extubated 4 days postinterventionally. Follow-up sonography 4 months postinterventionally showed dramatic regression of the hepatic tumor. We conclude that the coil-occlusion of hepatic hemangioendothelioma should be performed early in symptomatic newborns as arterial puncture and its associated complications can be avoided by using a venous approach with passage to the arterial side via the ductus arteriosus. In addition, in this age group, access to the portal system can be gained through the umbilical vein to occlude portal venous supplies to the hepatic hemangioendothelioma. PMID- 9857460 TI - [Increased acceptance of the International Normalized Ratio (INR) as a monitoring parameter of oral anticoagulation therapy in Germany. GELIA Study Group]. AB - In 1994 the German Cardiac Society recommended the International Normalized Ratio (INR) instead of "Quick's test" for the long-term management of oral anticoagulation therapy. Parallel cardiac surgical centers in Germany and Switzerland started patient recruitment for the GELIA study (German Experience with Low Intensity Anticoagulation). By the end of 1996, 862 consecutive patients with mechanical heart valve replacement had been enrolled. According to the protocol of the GELIA study, intensity of anticoagulation, methods used to control the oral anticoagulation therapy as well as other factors relevant for the anticoagulation management are reported every three months. The so far accumulated data material of the GELIA study seemed, therefore, appropriate to analyze potential changes in the acceptance of INR during the time period 1993 1996. Potential differences in the anticoagulation management were analyzed separately according to the time of enrollment of patients and changes during the follow up period, e. g., a switch from "Quick's test" to INR. Taking the time of enrollment into account, the percentage of patients who had INR-controlled oral anticoagulation therapy increased from 0% (1993), 14.3% (1994), 63.2% (1995) to 74.2% in 1996. During the follow up, there was a change from "Quick's test" to INR controls in 7 patients (1994; 2.1%), 67 patients (1995; 10.5%) and 142 patients (1996; 17.3%) with a consequent increase of INR controlled patients to a total of 34.2% in 1994, 52.0% in 1995, and 73.9% in 1996. PMID- 9857461 TI - [Intracardiac thrombosis and consecutive thromboembolisms in patients with heart valve diseases: predisposition and concepts for prevention]. AB - For patients with acquired heart valve lesions with increased risk for intracardiac thrombosis and consequent cardiogenic embolism there is consensus that oral anticoagulation therapy improves the overall prognosis. In mitral valve lesions anticoagulation is necessary after manifestation of atrial fibrillation or in cases of unstable sinus rhythm. The risk for thromboembolic events is increasing parallel to the enlargement of the left ventricular enddiastolic diameter, the left atrial size and dropping cardiac index. Spontaneous echo contrast (so called smoke like echos) indicate a prethrombotic state. In these cases an intensive anticoagulation is indicated. Aortic valve lesions require anticoagulation after manifestation of atrial fibrillation, the first manifestation of a thromboembolism or of spontaneous echo contrast. The risk for thromboembolism is increasing parallel to the reduction of left ventricular pump function. Life long oral anticoagulation therapy should be managed by use of the International Normalized Ratio (INR), and should be individualized taking into account patient related cardiac morphology and physiology, which may predispose to cardiogenic embolism. The target INR can range between 2.0 and 4.0. PMID- 9857462 TI - [Molecular mechanisms and indicators of thrombogenesis after heart valve replacement]. AB - Complex interactions among constituents of blood, components of the subendothelial extracellular matrix, and artificial surfaces of cardiovascular devices are involved in the thrombogenesis following heart valve replacement. Recently, the molecular basis of some of these interactions has been studied in detail. These insights have extended our understanding of interactive processes between platelet receptors, adhesive macromolecules, and abnormal flow conditions during platelet adhesion and aggregation. On the basis of new experimental data, it is concluded that circulating nonactivated platelets are capable of recognizing surface-bound fibrinogen via their glycoprotein receptor GPIIb-IIIa and become activated as a consequence of this specific interaction. In addition, a molecular mechanism has been proposed indicating that high-shear stress created by prosthetic heart valves can induce platelet aggregation. For this platelet reaction, multimeric von Willebrand factor is essential; its structure function relationship and its interaction with the platelet membrane GPIb receptor have been elucidated. The progress made provides specific targets for selective antithrombotic strategies. Moreover, new molecular activation markers have become available which permit appropriate control of established and experimental antithrombotic regimens. This paper reviews some aspects of recent advances in this area. PMID- 9857463 TI - [Thrombosis of prosthetic heart valves: diagnosis and management]. AB - The incidence of prosthetic valve thrombosis (PVT) has been reported to be < 0.1 0.25%/year following isolated aortic valve replacement and < 0.1-2.5%/year after mitral valve implantation. In a consecutive series 10.3% of PVT occurred after an interruption of oral anticoagulant therapy for weeks or months, in 20.7% with inadequate low anticoagulation while in 34.5% of PVT cases intensity of the oral anticoagulant therapy showed significant ups and downs. There was an obvious increase of PVT incidences during the winter months, concomitant with increasing fibrinogen levels and plasma viscosity. The diagnosis of PVT can be made with a high specificity and sensitivity by analysis of the typical opening and closing clicks of the valve occluder. Parameters indicating chronic intravascular hemolysis, fluoroscopy and echocardiography are additionally helpful to confirm the diagnosis. The therapy of choice is prosthetic valve re-replacement. Systemic thrombolytic therapy may be an alternative in selected patient groups. PMID- 9857464 TI - [Incidence of thromboembolism after heart valve replacement: modifying factors]. AB - The most important factors influencing thromboembolic rates after valve replacement are: 1) Cardiogenic factors such as left ventricular function, left atrial size and atrial rhythm. 2) Prosthesis-related factors such as type of prosthesis, position of prosthesis, and time after implantation. 3) Patient related factors such as quality of anticoagulation and gender. 4) Conditions with hypercoagulability. PMID- 9857465 TI - [Intensity of oral anticoagulant therapy after heart valve replacement]. AB - In recent years randomized clinical trials using standardized prothrombin time (international normalized ratio, INR) for the control of oral anticoagulant therapy have demonstrated that the use of a less intense therapeutic range decreases the rate of bleeding without reducing antithrombotic efficacy. Based on these results, the following guidelines for therapeutic ranges in heart valve replacement can be presently recommended: INR of 2.0-3.0 in patients with bioprosthetic valves for the first 3 months-except, probably, for patients with tissue valves in aortic position who are in sinus rhythmus; long-term anticoagulation corresponding to an INR of 2.0-3.0 in all patients with bioprosthetic valves and either atrial fibrillation or postoperative arterial embolism or a left atrial thrombus at surgery; long-term treatment equivalent to an INR of 2.5-3.5 in all patients with mechanical prosthetic heart valves. PMID- 9857466 TI - [Value of molecular markers for administration of anticoagulant therapy after heart valve replacement]. AB - In a prospective study, 39 patients following aortic or mitral valve replacement underwent investigation of molecular markers (prothrombin fragment F1+2, Factor II) in the initial phase of oral anticoagulation therapy (OAT). The results demonstrate that, despite INR being in the depicted range, the levels of the molecular markers remained high, indicating an increased risk of thromboembolic events. This leads to the conclusion that molecular markers are superior to INR in the monitoring of the early phase of OAT. PMID- 9857467 TI - [Inhibition of thrombocyte aggregation after heart valve replacement]. AB - After implantation of a mechanical cardiac valve, lifelong, full anticoagulation has been the tradition. After implantation of a biological valve, anticoagulation during 3 months subsequent to the operation is customary. This review evaluates the role of platelet inhibition after cardiac valve replacement. Platelet aggregation is inhibited effectively by aspirin in a daily dose of 100-160 mg. At this dose, episodes of severe bleeding are not significantly more frequent than during placebo, whereas patients on full anticoagulation bleed at a rate of 2% per year. After implantation of a mechanical cardiac valve, sole platelet inhibition is inferior to full anticoagulation. With a lower rate of bleeding, aspirin appears to prevent thromboembolic episodes caused by mechanical bileaflet prostheses (e.g., St. Jude) in the aortic position, and is almost as efficient as full anticoagulation. In Germany, patients with a porcine bioprosthesis, as currently in use, in the aortic position, frequently receive 100 mg aspiring per day. For patients with porcine bioprostheses in the mitral plus eventually in the aortic position in stable sinus rhythm, 100 mg aspirin per day is preferred to anticoagulation. For children with mechanical aortic valves, aspirin (2 mg/kg/day) needs to be considered an effective and convenient alternative to anticoagulation. Combining anticoagulation with 100 mg aspirin per day after implantation of a left-sided mechanical cardiac valve is pathophysiologically sound, but used to be considered as rendering the patients too bleeding-prone. Recently, total mortality and morbidity definitely have been demonstrated to be reduced by combined anticoagulation and platelet inhibition as compared to sole anticoagulation. Combining platelet inhibition with moderate anticoagulation (INR 2.0-3.0) was superior to combining it with full anticoagulation in terms of safety (INR > 3.0). Thus, current evidence favors combining moderate anticoagulation with 100 mg aspirin per day after left-sided mechanical cardiac valve replacement. PMID- 9857468 TI - [Anticoagulation in surgery, after hemorrhagic complications and in pregnancy]. AB - A review of the current management of the anticoagulated patient with heart valve prosthesis is presented. All patients with mechanical heart valve prostheses require life-long oral anticoagulation with coumarin or one of its derivatives. Recommendations for the prevention of thromboembolic events and bleeding complications in patients undergoing noncardiac operations are given. Furthermore, risk factors for hemorrhagic complications during long-term oral anticoagulation are discussed as well as management in case of acute bleeding. The advantages and disadvantages of three different treatment approaches to anticoagulation during pregnancy are discussed with delineation of specific risks for the mother and the fetus. PMID- 9857469 TI - [Anticoagulation in patients with heart valve replacement in pregnancy]. AB - Oral anticoagulants are the drugs of first choice for an effective anticoagulation after mechanical and biological heart valve replacement. Patients with mechanical heart valves are anticoagulated life long, whereas biological prostheses are anticoagulated for 3 months postoperatively in the case of sinus rhythm. Life-long anticoagulation in biological heart valve replacement is indicated if atrial fibrillation or dilatation of the left atrium or peripheral arterial embolism are present. During pregnancy oral anticoagulants are contraindicated in the first trimester. Heparins are effective alternatives for anticoagulation during this period. However, the dose of heparin has to be adjusted according to the activated partial thromboplastin time, which should be 1.5-fold prolonged. Low-molecular-weight heparins may be an alternative due to improved pharmacokinetics. Neither unfractionated nor low-molecular-weight heparin cross the placenta. PMID- 9857470 TI - [Patient self-monitoring in follow-up of long-term anticoagulant therapy]. AB - The CoaguChek system is a new system for simple self-testing of prothrombin time from capillary blood. The instrument works with a measuring principle which is completely new in coagulation: the motion of magnetic micro-particles in an oscillating magnetic field is stopped by the fibrin clot and is detected by a photodetector. The existing data show the analytical and clinical reliability of the system. Determinations for within series imprecision yielded a mean CV of 4.9%. The system is comparable to the comparison method Hepato Quick (y = 1.07 x 0.9% Quick; r = 0.89). Patient self-testing increases the relative time within therapeutic range from 54% to 84%. PMID- 9857471 TI - [Optimizing long-term anticoagulation by patient self-management?]. AB - An active role of the patient in managing his own therapy leads to an optimized quality of long-term anticoagulation and to a reduction of hemorrhagic or thromboembolic complications of treatment. Our study shows that patients could be easily trained in the self-management of oral anticoagulation and that they achieve superior anticoagulation control. In 10 years 1244 patients were trained in the self-management of oral anticoagulation at the Cardiac Rehabilitation Center in Bad Berleburg, Germany. With regard to their self-determined prothrombin times (PT) 387 patients could be followed up. The results were within the target range in 81.5% of the PT measurements taken by the patients themselves. Only 18.5% PT's were outside of the target range. The incidence of more important bleedings was 0.33% per year and of thromboembolic complications was 0.25% per year. Self-management of oral anticoagulation is associated with a reduced risk of hemorrhagic and thromboembolic complications and an improvement of prognosis and quality of life. PMID- 9857472 TI - A [[TiCl2(isodiCp)]2(mu-O)] dimer and a twinned cyclic [TiCl(isodiCp)(mu-O)]4 tetramer. AB - The mu-oxo dimer mu-oxo-bis[dichloro[(1,2,3,3a,7a-eta)- 4,5,6,7-tetrahydro-4,7 methano-1H-indenyl]titanium], [Ti2Cl4(mu-O)(C10H11)2], and the mu-oxo cyclic tetramer, cyclo-tetra-mu-oxo-tetrakis[chloro[(1,2,3,3a,7a-eta)-4,5,6,7-tetrahydro 4,7-methano-1H-indenyl]titanium], [Ti4Cl4(mu-O)4(C10H11)4], have been synthesized by electrophilic attack of TiCl4 on the exo-trimethylsilyl derivative of isodiCp (isodiCp is the isodicyclopentadienide anion or 4,5,6,7-tetrahydro-4,7 methano-1H-indene) in the presence of trace amounts of moisture. Both structures are endo isomers, with each Ti atom bonded in a eta 5 manner to the Cp ring of the isodiCp ligand. Both display a slight bending of the isodiCp group about the bond common to the Cp ring and the norbornane fragment, such that the norbornane group is displaced in the direction away from the Ti atom. The dihedral angles which describe this bending are 9.0 (3) and 10.2 (3) degrees for the dimer, and 9.2 (2) degrees for the tetramer. The dimer contains a bent Ti-O-Ti angle of 159.5 (2) degrees. The tetramer forms tetragonal crystals, which are twinned by merohedry on the (110) plane. The tetramer molecule contains a crystallographic 4 axis, with the Ti atoms linked by the O atoms in a mu-oxo manner to form an eight membered ring. PMID- 9857473 TI - 5-ammoniosalicylic acid chloride monohydrate. AB - The title compound, C7H8NO3+.Cl-.H2O, crystallized in the centrosymmetric space group P2(1)/c in an ionic form, the proton from HCl having been transferred to the amino N atom. The three H atoms on N, the H atom on the carboxyl group, the H atom of the hydroxyl group and the H atoms of the water molecule, all of which are involved in hydrogen bonding, are ordered. The eight 'best' hydrogen bonds have the following donor-acceptor distances: O...O 2.681 (2) and 2.598 (2); O...Cl 3.367 (1), 3.345 (2) and 3.156 (2); N...O 2.995 (2); N...Cl 3.173 (2) and 3.114 (2) A. In this structure, the acid cations are not linked directly to each other by hydrogen bonds, but are linked indirectly, via hydrogen bonds involving chloride ions and water molecules, into a three-dimensional network. Through basic second-level graphs, finite patterns substantially outnumber chains and rings. PMID- 9857474 TI - (4R)-(-)-2-thioxothiazolidine-4-carboxylic acid (raphanusamic acid). AB - The title acid, C4H5NO2S2, crystallized in the noncentrosymmetric space group P2(1)2(1)2(1) with one molecule as the asymmetric unit. Two hydrogen bonds occur, namely, N--H...O, with a donor-acceptor distance of 2.850 (2) A, and O-H...S (resonance induced), with a donor-acceptor distance of 3.318 (2) A; both H atoms involved are ordered. These two types of bonds link a given molecule to four neighbors, the molecules linked being of space-group symmetry types 1 and 3, or 2 and 4. These two subsets of molecules alternate in layers along c. Three significant C-H...O interactions occur, one of which crosslinks the subsets just described to produce a three-dimensional network of linked molecules. The absolute structure is determined. PMID- 9857475 TI - N-borane-N-(trimethylsilyl)morpholine. AB - The title compound, C7H20BNOSi, features an N,N-disubstituted six-membered morpholine ring in a chair conformation, with the trimethylsilyl group in the equatorial position and the borane group in the axial position. The least-squares plane formed by the four C atoms of the morpholine ring has a mean deviation of 0.013 (2) A. The O and N atoms are 0.672 (2) and 0.650 (2) A above and below the plane, respectively. PMID- 9857476 TI - The antifungal drug clotrimazole. AB - The structure of the title compound, 1-[(2-chlorophenyl)diphenylmethyl]-1H imidazole, C22H17ClN2, has been determined. The molecular conformation showed a weathercock-type structure and the three phenyl rings are almost perpendicular to the imidazole ring. The distances between the centres of the three phenyl rings and the centre of the imidazole ring are in the range 4.52-4.54 A. PMID- 9857477 TI - Propagation of a signal coordinating force generation along an actin filament in actomyosin complexes. PMID- 9857478 TI - Interactions of surface-confined DNA with acid-activated mitomycin C. AB - The anti-cancer drug mitomycin C (MC) was acid-activated and its interaction with single-stranded calf thymus DNA, immobilized at the surface of the hanging mercury drop electrode (DNA-modified HMDE) was studied by cyclic voltammetry. It was found that immersion of the DNA-modified electrode in a solution of acid activated MC (at pH 3.9) for a short time (usually 1 min) at open current circuit, followed by transfer of the electrode in a neutral blank background electrolyte, resulted in a decrease of the anodic peak G (due to guanine residues in DNA) and in the formation of a reversible couple at approx. -0.44 V. The potential of the cathodic peak was approx. 50 mV more negative than the cathodic peak of the acid-activated MC obtained under the same conditions in the absence of DNA. No changes of peak G occurred and only a very small cathodic peak appeared if the DNA-modified electrode was immersed in an MC solution not exposed to acid pH. On the basis of these results and additional experiments, including dependence on concentration, time and pH during the interaction of MC with DNA at the electrode surface, we concluded that acid-activated MC is covalently bound to guanine residues in DNA immobilized at the electrode surface and that the quinone group in the DNA-MC adduct is reversibly reduced at the electrode. PMID- 9857479 TI - Differential stability of E. coli ribosomal particles and free RNA towards thermal degradation studied by microcalorimetry. AB - We investigated the thermal degradation of E. coli ribosomes by differential scanning microcalorimetry. The 70S particles show two distinctive and irreversible peaks upon thermal degradation. Free rRNA in solution produces, on the contrary, an unstructured denaturation profile. The thermal analysis of 50S particles shows a profile substantially identical to that observed in 70S, while 30S particles produce an unstructured denaturation pattern. Therefore the thermal behavior of the 70S particle is essentially attributable to the denaturation of the 50S subunit. Our data validate previous observations that the 50S has a more rigid structure as compared to 30S, which behaves as a 'floppy' particle. In addition our data suggest that protein/RNA interactions play a significant role to stabilize three-dimensional structures of the ribosome. PMID- 9857480 TI - Analysis of hydrophobic and charged patches and influence of medium- and long range interactions in molecular chaperones. AB - The amino acid composition of the aromatic residues Phe, Tyr and Trp are much less significant in chaperones and the residues Cys, Glu, His, Met and Pro vary significantly in chaperones compared to normal globular proteins. In the present work, we have analysed the hydrophobic and charged patches in molecular chaperones which provide more insight for a better understanding of chaperone folding. Also, we have investigated the role of medium- and long-range contacts in chaperones and the preference of amino acid residues influenced by these interactions. Furthermore, the role of hydrophobic and helix-forming residues and disulfide bonding in these interactions have been discussed. PMID- 9857481 TI - The relationship between biological activity and the electronic structure and transfer of the whole acidic PLA2 molecule in ab initio level. AB - The electronic structure of the whole molecule of acidic phospholipase A2 (PLA2) from the venom of Agkistrodon halys pallas (A. halys pallas) has been calculated using the extended negative factor counting (ENFC) method in which dimers were calculated at the ab initio level using a minimal basis set, with simulation of the aqueous environment. Hopping conductivities were determined by the use of random walk theory. The results show that the frontier orbitals are mainly localized to residues which are involved in the biological activity of acidic PLA2. The C-terminal region might play some important role in biological activity because of its active electrons. The aromatic patch on the surface of the enzyme, together with two neighbouring acidic residues, has very active electrons that may be responsible for the inhibition of platelet aggregation. Trp30, which is involved in the interfacial recognition region, may transfer its electrons to the aggregated substrate. It is also concluded that the conductivity of the protein is caused mainly by holes transported through the valence band rather than electrons transferred in the conductive band. The a.c. conductivity of acidic PLA2 confirms that proteins, if doped, are amorphous conductors. Moreover, the a.c. conductivities of acidic PLA2 are approximately one order of magnitude higher than those of some other proteins. This suggests that the toxicity of acidic PLA2 may be related to its high a.c. conductivity. PMID- 9857482 TI - Hemoglobin-oxygen-carbon monoxide equilibria with the MWC model. AB - Fractional saturation equations for the Monod, Wyman and Changeux model are derived for the case of two distinct ligands bonding to a host molecule with four ligand sites and two conformational states. A variety of useful graphical studies can be derived from these equations when applied to normal human hemoglobin with O2 and CO as ligands. For example, the oxygen transport capability of hemoglobin can be assessed at different environmental CO levels and the concentrations of various liganded species can be displayed as a function of fractional saturation with oxygen. In addition, the CO pressure in the tissue, PCOtissue, can be calculated as a function of the tissue oxygen pressure, PO2tissue, at different environmental levels of CO. In an environment of a given CO concentration, PCOtissue decreases with PO2tissue until a minimum is reached. Further decrease in PO2tissue results in a fairly steep rise in PCOtissue. PMID- 9857483 TI - Mechanism of the alpha-complementation reaction of E. coli beta-galactosidase deduced from fluorescence correlation spectroscopy measurements. AB - The kinetics of the alpha-complementation reaction of two protein fragments yielding active E. coli beta-galactosidase was measured using fluorescence correlation spectroscopy (FCS). The association reaction was extremely slow with an apparent association rate kapp of 207 M-1 s-1. This low association rate can be explained by a fast pre-equilibrium and slow subsequent steps involving at least two dimeric complexes. The subsequent formation of a tetrameric complex is probable and consistent with the experimental data. The complexes comprise two or four subunits, respectively, of the large fragment (EA)2 and in all cases only one small fragment, ED which has been labeled with Cy5. These kinetics have been compared to the association kinetics of ED to inactivated (EA)2. The kinetics were similar to the association with native (EA)2. The data support the observation that lyophilization of (EA)2 in a reducing environment which causes complete loss of enzymatic activity does not interfere with binding. PMID- 9857484 TI - Blood plasma levels of lipoperoxides, glutathione peroxidase, beta carotene, vitamin A and E in women with habitual abortion. AB - The plasma levels of lipoperoxides, glutathione peroxidase (GSH-Px), reduced glutathione (GSH), beta carotene, vitamin A, E, some plasma biochemical and blood haematological parameters were investigated in 40 women with habitual abortion (HA) and controls. The levels of GSH, vitamin A, E and beta carotene were significantly lower in women with HA than in controls. However, the plasma levels of lipid peroxidation, alkaline phosphatase (ALP), glucose and blood haemoglobin were significantly higher in HA than in controls. In addition, plasma levels of GSH-Px, AST, ALT, total bilirubin, total protein, albumin, sodium, potassium, calcium and number of white blood cells, red blood cells, platelet and values of packet cell volume showed no significant differences between HA and controls. According to the results of this study, we observed that the levels of lipid peroxidation were increased and plasma levels of vitamin A, E and beta carotene were decreased in HA. The decrease of those antioxidants may play a significant role in women with habitual abortion. PMID- 9857485 TI - Failure of streptozotocin tetraacetate to undergo extensive hydrolysis and to inhibit D-glucose metabolism and insulinotropic action in rat pancreatic islets. AB - The esterification of several monosaccharides, such as D-glucose, D mannoheptulose and 2-deoxy-D-glucose was recently reported to increase their biological efficiency as either nutrient or antimetabolic agent. In the present study, however, the tetraacetate ester of streptozotocin was unexpectedly found to be less potent than unesterified streptozotocin in inhibiting D-glucose metabolism and insulinotropic action in isolated rat pancreatic islets. This coincided with a much lower rate for the hydrolysis of streptozotocin tetraacetate than D-glucose pentaacetate in islet homogenates. These findings document that the esterification of single sugars is not always a successful procedure to enhance their biological potency, for instance because of too low a rate for the intracellular hydrolysis of the ester. To the extent that the activity of the concerned esterase(s) may differ in distinct cell types, as suggested by a prior observation, advantage could be taken of such a situation to target selected esters towards specific, e.g. tumoural cells. PMID- 9857486 TI - DNA repair, cell cycle progression and cell death following camptothecin treatment in two murine lymphoma L5178Y sublines. AB - The processes involved in cell response to camptothecin (CPT) were investigated in two sublines of L5178Y (LY) murine lymphoma; LY-R, resistant and LY-S, sensitive to X-irradiation, which are inversely cross-sensitive to the drug. The cells were pulse-treated with 2 microM CPT for 1 h; this resulted in equal numbers of replication-related DNA double-strand breaks (DSBs) in both sublines. After drug removal, at different time points up to 24 h, the levels of DSBs were measured by using field inversion gel electrophoresis (FIGE) and comet assay at neutral pH. Both methods revealed faster DSBs repair in LY-S than in LY-R cells, in contrast with X-ray-induced DSBs. This however, was followed by the appearance of secondary breaks in the former subline. The cell cycle arrest was at S/G2 phase and comprised equal numbers of cells in LY-S and LY-R populations. In both sublines formation of giant cells took place, as well as delayed apoptosis starting about 20 h post-CPT incubation and proceeding with similar intensity. At the same time, the total number of necrotic cells appearing during post-exposure incubation in the LY-R subline exceeded that in the LY-S subline. We suggest that, beside previously documented higher susceptibility of topoisomerase I (Topo I) from LY-R cells to CPT, a higher initial rate of replication-related DSBs repair, but not lower propensity to apoptosis, may contribute to the relative CPT resistance of LY-S versus LY-R cells. PMID- 9857487 TI - Effect of dietary fat on lymphocyte proliferation and metabolism. AB - The effect of diets enriched with fat containing different fatty acids on glucose and glutamine metabolism of mesenteric lymph nodes lymphocytes, spleen, and thymus and lymphocyte proliferation was examined. The following fat-rich diets were tested: (1) standard chow (CC); (2) medium chain saturated fatty acids (MS)- coconut fat oil; (3) long chain saturated fatty acids (LS)--cocoa butter; (4) monounsaturated fatty acids (MU)--canola oil (n-9); (5) polyunsaturated fatty acids (PU)--soybean oil (n-6). Of the fat-rich diets tested, MS was the one to present the least pronounced effect. Lymphocyte proliferation was reduced by LS (64 per cent), MU (55 per cent), and PU (60 per cent). Hexokinase activity was enhanced in lymph node lymphocytes by PU (67 per cent), in the spleen by MS (42 per cent), and in the thymus by PU (30 per cent). This enzyme activity was reduced in the spleen (33 per cent) by LS and MU (35 per cent). In the thymus, this enzyme activity was reduced by LS (26 per cent) and MU (13 per cent). Maximal phosphate-dependent glutaminase activity was raised in lymphocytes by MS (70 per cent) and MU (20 per cent). This enzyme activity, however, was decreased in lymphocytes by PU (26 per cent), in the spleen by LS (15 per cent), and in the thymus by MU (44 per cent). Citrate synthase activity was increased in lymphocytes by MU (35 per cent), in the spleen by LS (56 per cent) and MU (68 per cent), and in the thymus by LS (42 per cent). This enzyme activity was decreased in lymphocytes by PU (24 per cent) only. [U-14C]-Glucose decarboxylation was raised by all fat-rich diets; MS (88 per cent). LS (39 per cent), MU (33 per cent), and PU (50 per cent), whereas [U-14C]-glutamine decarboxylation was increased by LS (53 per cent) and MU (55 per cent) and decreased by MS (17 per cent). The results presented indicate that the reduction in lymphocyte proliferation due to LS, LU and PU could well be a consequence of changes in glucose and glutamine metabolism. PMID- 9857488 TI - Oxidative activity in mitochondria isolated from rat liver at different stages of development. AB - The purpose of this study was to evaluate the oxidative capacities in hepatic mitochondria isolated from prepubertal, young adult and adult rats (40, 90 and 180 days of age, respectively). In these rats, mitochondrial respiratory rates using FAD- and NAD-linked substrates as well as mitochondrial protein mass were measured. The results show that only the oxidative capacity of FAD-linked pathways significantly declined in mitochondria from 180-day-old rats compared with those from younger animals. When we consider FAD-linked respiration expressed per g liver, no significant difference was found among rats of different ages because of an increased mitochondrial protein mass found in 180 day-old rats. However, when FAD-linked and lipid-dependent respiratory rates were expressed per 100 g body weight, significant decreases occurred in 180-day-old rats. Therefore, the decrease in liver weight expressed per 100 g body weight rather than an impaired hepatic cellular activity may be the cause of body energy deficit in 180-day-old rats. PMID- 9857489 TI - Exercise, depletion of antioxidants and antioxidant manipulation. AB - Strenuous physical activity is known to increase the production of reactive oxygen species (ROS), associated with depletion of antioxidant defence. In the present work we evaluated the level of lipid peroxidation and antioxidant components in blood of sportsmen under resting conditions and compared the data obtained with those in age- and sex-matched sedentary controls. A significant increase was noted in the levels of thiobarbituric acid reactive substances (TBARS) and conjugated dienes while a decrease was observed in ascorbic acid and glutathione levels in sportsmen. alpha-Tocopherol was unaltered in plasma of sportsmen as compared to controls. The activity of superoxide dismutase was increased (52 per cent) and glutathione peroxidase was decreased (43 per cent) in the erythrocytes of sportsmen compared to controls. Basal glutathione levels were negatively correlated with conjugated dienes and maximal oxygen uptake (VO2max) of the subjects. Dietary supplementation with antioxidant vitamins has been shown to be beneficial in combating oxidative stress without enhancing performance while exogenous glutathione was found to influence the endurance capacity of athletes. Such studies demonstrate the critical role played by glutathione and suggest that intervention trials should include a mixture of antioxidants rather than a single antioxidant. PMID- 9857490 TI - Alteration of phosphotyrosine-containing proteins during differentiation of chicken erythroleukemia cells (HD3). AB - After treatment of HD3 cells with erythroid-inducing agents (hemin and butyric acid) at 42 degrees C, the profile of phosphotyrosine-containing proteins was altered. Upon induction the overall level of phosphotyrosine-containing proteins increased. To examine the role of protein phosphorylation in HD3 cells differentiation, the cells were treated with specific inhibitors. In the presence of okadaic acid, cell proliferation was arrested and accompanied by a marked increase in haemoglobin synthesis, a differentiation marker of erythroid cells. Okadaic acid caused decrease of the phosphotyrosine-containing proteins, presumably to maintain a balance between phosphorylation/dephosphorylation processes in the cells. Addition of 3-isobutyl-1-methyl-xanthine, an activator of phosphatases, caused a decrease or disappearance of almost all phosphotyrosine containing proteins and, at the same time, prevented the erythroid differentiation of HD3 cells. Sodium orthovanadate, a specific inhibitor of phosphotyrosine phosphatase, increased the level of phosphotyrosine proteins and induced differentiation of HD3 cells. These results indicate that phosphorylation of cellular proteins is coupled with a reaction(s) which is responsible for triggering the differentiation of HD3 cells. The phosphorylation/dephosphorylation processes are associated with an early event(s) during the differentiation of HD3 cells and may not be connected to tyrosine residues. PMID- 9857491 TI - Methotrexate: pentose cycle and oxidative stress. AB - The effect of methotrexate (MTX) and leucovorin (LCV) on pentose cycle enzymes and the activity of enzymes involved in enzyme defence mechanisms against ROS in HeLa cells, were studied. The effect of MTX was also investigated on the cellular levels of glutathione. MTX inhibited the activity of glucose-6-phosphate and 6 phosphogluconate dehydrogenases. The activities of glutathione reductase and gamma-glutamylcysteine synthetase were also inhibited by the drug. No effect was observed on the activities of catalase, superoxide dismutase or transketolase. LCV had no effect on any of the enzymes studied. MTX decreased the cellular levels of glutathione (70 per cent), while the presence of LCV and glutamine did not interfere with the effect of MTX. The net results appear to show that the biological situation resulting from treatment with MTX leads to a reduction of effectiveness of the antioxidant enzyme defence system. PMID- 9857492 TI - Referential communication and category acquisition. AB - Explanations of category coherence include that categories reflect feature correlations in the world, that the human conceptual system is designed to create systematic categories, and that people have theories about the world that bind together seemingly unrelated features. The authors have suggested that the need to establish reference in communication also influences category coherence. This proposal was tested in 2 studies involving a referential communication task. In these studies, consistency was promoted between individuals by communication, which synchronized the category structures of different people. Further, people were focused on the commonalities of objects and on the differences related to the commonalities by communication--a pattern that is compatible with what has been observed in existing categories. These results suggest that categorization research must incorporate communication tasks into the canon of methodologies used to study category structure. PMID- 9857493 TI - Conscious and unconscious strategy discoveries: a microgenetic analysis. AB - We examined second graders' strategy discoveries on a task that could be solved either by arithmetic computation or by a simple insight. Strategy use was assessed on each trial through both an implicit measure and an explicit one. The results provided unambiguous evidence that strategies can be discovered without conscious awareness. Almost 90% of children showed the insight at an implicit, unreportable level before they showed it at an explicit, reportable one. When children received the relevant problems on each trial, 80% reported the insight within 5 trials of when their solution times indicated that they first generated it. Implications for understanding the relation between unconscious and conscious discoveries and for the role of competing activations in strategy discovery are discussed. PMID- 9857494 TI - Does consistent scene context facilitate object perception? AB - The conclusion that scene knowledge interacts with object perception depends on evidence that object detection is facilitated by consistent scene context. Experiment 1 replicated the I. Biederman, R. J. Mezzanotte, and J. C. Rabinowitz (1982) object-detection paradigm. Detection performance was higher for semantically consistent versus inconsistent objects. However, when the paradigm was modified to control for response bias (Experiments 2 and 3) or when response bias was eliminated by means of a forced-choice procedure (Experiment 4), no such advantage obtained. When an additional source of biasing information was eliminated by presenting the object label after the scene (Experiments 3 and 4), there was either no effect of consistency (Experiment 4) or an inconsistent object advantage (Experiment 3). These results suggest that object perception is not facilitated by consistent scene context. PMID- 9857495 TI - Use of the Hand Test in the classification of psychiatric inpatient adolescents. AB - This study investigates the extent to which 2 Hand Test variables, the Pathology summary score (PATH) and the Acting Out Score (AOS), were able to differentiate among 3 groups of adolescents. Thirty psychiatric inpatient adolescents diagnosed with various Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994) behavioral and emotional disorders were compared to 30 outpatient and 30 nonclinical adolescents matched on age, grade, and sex. An analysis of variance demonstrated the ability of both PATH and AOS to significantly differentiate among the 3 groups. Multiple regression equations utilized PATH and AOS as significant predictors of inpatient status (p < .0001) and general patient (inpatient and outpatient) status (p < .0001). Utilizing diagnostic efficiency statistics, overall correct classification was found to be highest for PATH > or = 3 and AOS > or = 2. These findings support the application of the Hand Test as a useful assessment tool for adolescents suspected of having psychiatric disorders and present evidence for the use of PATH and AOS for classification purposes in ways that are clinically meaningful. PMID- 9857496 TI - The diagnostic meaning of "nervous breakdown" among lay populations. AB - Two studies investigated the nature of nervous breakdown (NB): a mental illness commonly referred to by individuals not professionally related to the field of mental health. The clinical profile of NB was first surveyed among 121 undergraduates using a symptom checklist. Descriptive statistics and analysis of variance indicated NB is a time-limited condition that presents with primarily anxious and depressed features, associated with a series of external precipitating stressors (e.g., interpersonal, employment, and financial losses). Dimensions significantly uncharacteristic of NB included psychoticism, somatization, phobic anxiety, and mania. These results were replicated in a community sample of 189 adults from urban and suburban areas of a large metropolitan city. Respondents held a relatively unitary view of NB: Combined sample cluster analysis (N = 310) revealed 2 groups with similar symptom profiles that differed primarily with regard to level of distress associated with disorder. Thus, among this population, NB is not an amorphous term for generalized psychiatric disturbance. PMID- 9857497 TI - Predicting back pain sufferers' intentions to exercise. AB - One important factor in reducing back pain is the performance of exercises prescribed by physicians during treatment. The present investigation was conducted to examine the correlates of an individual's intention to adhere to such prescriptions. On the bases of M. Fishbein's (1980) theory of reasoned action and I. Ajzen's (1988) theory of planned behavior, attitudes, subjective norms, and 2 types of perceived behavioral control were measured in a sample of American patients. Attitudes and subjective norms failed to predict intentions to exercise in accordance with the physician's orders, but each of the perceived behavioral control measures did moderately well as an independent predictor and quite well when combined in a multiple regression approach. PMID- 9857498 TI - On gender specificity of person characteristics in personnel advertisements: a study among future applications. AB - This research is an examination of how person characteristics mentioned in personnel advertisements influence the judgments of male and female students with regard to their subjective eligibility for, and the attractiveness of, a particular vacancy. The first study showed that many frequently mentioned person characteristics in personnel advertisements applied differently to men and women, but that female characteristics were more in demand than male characteristics. The second study examined to what degree male and female participants felt that they themselves possessed male and female characteristics; results showed that the men were more confident about the degree to which they possessed male and female characteristics than the women were. The third study examined how the type of person characteristics mentioned in personnel advertisements influenced men's and women's judgments regarding their eligibility for and the attractiveness of a particular position. The men found all positions about equally attractive, whereas the women found "male" positions considerably less attractive. Implications of the study are discussed. PMID- 9857499 TI - Assertive, offensive, protective, and defensive styles of self-presentation: a taxonomy. AB - Classifications of self-presentational behavior are reviewed for differences and similarities. Earlier classifications and recent empirical research are used as a basis for a taxonomy of four styles of self-presentation--assertive, offensive, protective, and defensive--according to the relevant self-presentational intentions and the level of activity involved. Each style is illustrated with typical behavioral examples, together with individual differences and possible consequences. PMID- 9857500 TI - Profiles of time perspective and personality: developmental considerations. AB - Time perspective is an important but subtle cognitive construct underlying personality, decision making, and goal setting. This study identified 3 temporal dimensions--temporal extension, temporal attitude, and temporal structure--and reviewed the associations among them. T. J. Cottle's (1969, 1977) work on temporal profiles was briefly reviewed; it suggested that 3 types of temporal profiles can be isolated and that 3 broad personality dimensions--human agency, mood, and temperament--from characteristic associations with each of the temporal profiles. The authors expected the profiles to reflect developmental sensitivities as a person moves from adolescence into early adulthood. Two samples (159 high school students and 236 university students) participated in the study. The results provided some limited support for the actualizer and atomist profiles; somewhat surprisingly, the role of temporal extension appeared to be insignificant. No evidence was found for a developmental transition of time perspective between middle adolescence (15-17 years) and early adulthood (18-25 years). PMID- 9857501 TI - Cognitive correlates of boredom proneness: the role of private self-consciousness and absorption. AB - The contributions of private self-consciousness and absorption in explaining boredom proneness were investigated. University students enrolled at a public university in the southeastern United States completed a packet containing the Boredom Proneness Scale (BPS; R. Farmer & N. D. Sundberg, 1986), the Self Consciousness Scale (SCS; A. Fenigstein, M. F. Scheier, & A. H. Buss, 1975), the Tellegen Absorption Scale (TAS; A. Tellegen & G. Atkinson, 1974), the Need for Cognition Scale (NCS; J. T. Cacioppo, R. E. Petty, & C. F. Kao, 1984), and a demographic questionnaire. Scores on the Boredom Proneness subscale, Internal Stimulation, which indicates the difficulty in keeping oneself interested and entertained, were significantly lower for individuals high in absorption (a measure of attention). Individuals high in positive self-awareness, representing awareness of one's internal states, reported lower overall boredom. Individuals high in negative self-awareness, which focuses on evaluation and judgment, reported increased total boredom proneness scores. Implications of these findings for the treatment of boredom proneness and future research are discussed. PMID- 9857502 TI - The accuracy of dysphoric and nondepressed groups' predictions of life events. AB - The phenomenon of depressive realism was examined in relation to the future prediction of positive and negative life events. A group of dysphoric (n = 20) and nondepressed (n = 38) British undergraduates participated in a prospective study lasting 3 months. Partly consistent with the depressive realism hypotheses, dysphoric participants were more realistic concerning the negative life events they would experience, but they were less realistic concerning the negative life events they would not experience. Although no difference was found for predicting the occurrence of positive life events, dysphoric participants were found to be more realistic concerning positive life events that they would not experience. PMID- 9857503 TI - Cross-cultural similarities in the personality dimensions of heroin users. PMID- 9857504 TI - Perception of microphone noise in hearing instruments. AB - In a well-designed instrument the noise level is reduced until it is dominated by the front end noise. In a hearing instrument this is the microphone noise. This paper examines the perception of noise in a hearing instrument by both normal hearing and hearing-impaired listeners. The noise levels are specified as input referred values in one-third octave bands. Two sets of measurements, the just objectionable level (JOL) and the just-audible level (JAL), were assessed in third octave bands from 250 to 5000 Hz. The data indicated that the use of a subjectively described acceptability of noise, JOL, is an unreliable measure due to a very large variability across listeners. It is recommended that mean values for the noise threshold level, the JAL, be used as a guide in the optimization of microphone noise design. PMID- 9857505 TI - Exposure-response relationships for transportation noise. AB - This article presents synthesis curves for the relationship between DNL and percentage highly annoyed for three transportation noise sources. The results are based on all 21 datasets examined by Schultz [J. Acoust. Soc. Am. 64, 377-405 (1978)] and Fidell et al. [J. Acoust. Soc. Am. 89, 221-233 (1991)] for which acceptable DNL and percentage highly annoyed measure could be derived, augmented with 34 datasets. Separate, nonidentical curves were found for aircraft, road traffic, and railway noise. A difference between sources was found using data for all studies combined and for only those studies in which respondents evaluated two sources. The latter outcome strengthens the conclusion that the differences between sources cannot be explained by differences in study methodology. PMID- 9857506 TI - Measurements and model of the cat middle ear: evidence of tympanic membrane acoustic delay. AB - In order to better understand the mechanics of tympanic membrane (TM) transduction at frequencies above a few kHz, the middle-ear (ME) impedance measured near the tympanic membrane is studied for three anesthetized cat ears after widely opening the ME cavities (MEC). Three conditions were measured: intact ossicles, drained cochlea, and disarticulated stapes. When the cochlear load is removed from the ME by disarticulating the stapes, the impedance magnitude varies by about +/- 25 dB in the 5- to 30-kHz range, with peaks and valleys at intervals of approximately 5 kHz. These measurements suggest middle ear standing waves. It is argued that these standing waves reside in the TM. In contrast, the magnitude of the impedance for the intact case varies by less than +/- 10 dB, indicating that for this case the standing waves are damped by the cochlear load. Since the measurements were made within 2 mm of the TM, standing waves in the ear canal can be ruled out at these frequencies. Although the ME cavities were widely opened, reflections from the ME cavity walls or surrounding structures could conceivably result in standing waves. However, this possibility is ruled out by model predictions showing that such large standing waves in the ME cavity space would also be present in the intact case, in disagreement with the observation that standing waves are damped by cochlear loading. As a first order approximation, the standing waves are modeled by representing the TM as a lossless transmission line with a frequency-independent delay of 36 microseconds. The delay was estimated by converting the impedance data to reflectance and analyzing the reflectance group delay. In the model the ossicles are represented as lumped-parameter elements. In contrast to previous models, the distributed and lumped parameter model of the ME is consistent with the measured impedance for all three conditions in the 200-Hz to 30-kHz region. Also in contrast with previous models, the ear-canal impedance is not mass dominated for frequencies above a few kHz. Finally, the present model is shown to be consistent, at high frequencies, with widely accepted transfer functions between (i) the stapes displacement and ear-canal pressure, (ii) the vestibule pressure and ear-canal pressure, and (iii) the umbo velocity and ear-canal volume velocity. An improved understanding of TM mechanics is important to improve hearing aid transducer design, ear-plug design, as well as otoacoustic emissions research. PMID- 9857507 TI - Low-level steady-state auditory evoked potentials: effects of rate and sedation on detectability. AB - Steady-state auditory evoked potentials (SSAEPs) in alert adults are most detectable at stimulus or modulation rates of about 40 Hz. Sedation reduces the detectability of 40-Hz SSAEPs and increases it for higher rate SSAEPs. This study examined whether rates higher than 40 Hz would be preferable for detecting responses to low-intensity tones in sedated adults. Fourteen normal adults listened to 640-Hz tones at modulation rates (and toneburst rates) of 20-160 Hz, in 10-Hz steps, at levels of 38 and 58 dB peak equivalent sound-pressure level (peSPL) (20 and 40 dB normal hearing level (nHL) for amplitude-modulated (AM) tones), both alert and sedated (1-2 g chloral hydrate). Sedation reduced both signal (SSAEP) power and noise power at all rates, but noise power reduction was greater for higher rates. Detectability in the alert condition was always greatest at 40 Hz. Under sedation, a second detectability peak was present at 90 Hz for 58-dB peSPL tones, approximately equal to that seen at 40 Hz. At 38 dB peSPL (sedated), peak detectability moved from 40 to 50 Hz. These results suggest that presentation/modulation rates around 40 Hz may be optimal for SSAEP detectability at low levels in adults, whether alert or sedated. PMID- 9857508 TI - Nonmonotonicity of informational masking. AB - Neff and Green [Percept. Psychophys. 41, 409-415 (1987)] report that the masking of single tones by random-frequency multitone maskers varies nonmonotonically with number of masker components (peaking at 10-50 components). In this paper it is shown that such results are well predicted by a model (the component-relative entropy model, CoRE) wherein thresholds increase linearly with the ensemble variance of masker spectra smoothed by peripheral auditory filters [R. A. Lutfi, J. Acoust. Soc. Am. 94, 748-758 (1993)]. Three experiments were conducted. In the first, the nonmonotonic relation was replicated for 9 of 11 listeners in conditions similar to those of Neff and Green. In the second, the frequencies of masker components were fixed and the levels of components were varied randomly across presentations to simulate Gaussian noise. In this case, the nonmonotonicity and the total amount of masking for these listeners were shown to be significantly reduced. In the third experiment, masked thresholds for the signal were found to vary monotonically with the frequency spacing of masker components for a fixed number of masker components. Large individual differences among listeners were obtained in some experimental conditions. Individual as well as mean thresholds were well predicted by the CoRE model with an appropriate selection of the values of the two free parameters of the model for each listener. PMID- 9857509 TI - Suppression and the upward spread of masking. AB - The purpose of this study is to clarify the role of suppression in the growth of masking when a signal is well above the masker in frequency (upward spread of masking). Classical psychophysical models assume that masking is primarily due to the spread of masker excitation, and that the nonlinear upward spread of masking reflects a differential growth in excitation between the masker and the signal at the signal frequency. In contrast, recent physiological studies have indicated that upward spread of masking in the auditory nerve is due to the increasing effect of suppression with increasing masker level. This study compares thresholds for signals between 2.4 and 5.6 kHz in simultaneous and nonsimultaneous masking for conditions in which the masker is either at or well below the signal frequency. Maximum differences between simultaneous and nonsimultaneous masking were small (< 6 dB) for the on-frequency conditions but larger for the off-frequency conditions (15-32 dB). The results suggest that suppression plays a major role in determining thresholds at high masker levels, when the masker is well below the signal in frequency. This is consistent with the conclusions of physiological studies. However, for signal levels higher than about 40 dB SPL, the growth of masking for signals above the masker frequency is nonlinear even in the nonsimultaneous-masking conditions, where suppression is not expected. This is consistent with an explanation based on the compressive response of the basilar membrane, and confirms that suppression is not necessary for nonlinear upward spread of masking. PMID- 9857510 TI - Profiling the perceptual suppression of partials in periodic complex tones: further evidence for a harmonic template. AB - The basis for the perceptual cohesion of periodic complex tones was investigated. In experiment 1, 2-4 consecutive components (harmonics 6 and above) were removed from a 14-harmonic complex and replaced with a sinusoidal "probe," located at one of a set of regularly spaced positions spanning the gap. On each trial, subjects heard a complex tone followed by an adjustable pure tone in a continuous loop. Subjects were better able to match the pure tone to the probe when the probe did not coincide with a harmonic position. Minima in "hit rate" were more pronounced when harmonic probes were in positions adjacent to other harmonics than when they were not. These findings suggest that the pitch of each in-tune partial was actively suppressed by a template whose influence attenuated with distance from regions of consecutive harmonics. In experiment 2, the partials on either side of the spectral gap were harmonics of different fundamental frequencies. Hit-rate minima corresponding to both fundamentals were found, indicating an upward and downward spread of suppression, and also demonstrating the concurrent operation of two templates. The results confirm recent findings in support of template models, and are consistent with the idea that partial-pitch suppression underpins harmonic fusion. PMID- 9857512 TI - Effects of ear of entry and perceived location of synchronous and asynchronous components on mistuning detection. AB - Listeners were required to detect mistuning imposed on the center ("target") component of a 200-ms complex consisting of the first seven harmonics of a 500-Hz fundamental. In the standard interval of each 2IFC trial, all components were frequency modulated in-phase by a 5-Hz sinusoid. In the signal interval the frequency modulation of the target component was inverted in-phase, thereby introducing a mistuning proportional to the depth of FM. In a similar experiment, using monaural presentation, Carlyon [J. Acoust. Soc. Am. 95, 2622-2630 (1994)] reported a substantial elevation of thresholds in the presence of an unmodulated asynchronous interferer with frequency identical to the mean frequency of the target. This was attributed to the interferer, causing the target component to be perceptually segregated from the remainder of the complex, thereby impairing across-frequency comparisons. Experiment 1 of the present study showed that an interferer presented contralaterally for 200 ms before and 100 ms after the signal complex (no simultaneous presentation) also impaired performance, but to a lesser extent than an ipsilaterally presented one. Experiment 2 showed that an interferer which was presented dichotically with an interaural level difference (ILD) of 10 dB, so that it was perceived contralaterally, had the same (large) effect as if it were presented ipsilaterally. Experiment 3 showed that, in the absence of any interferer, performance was impaired when the nontarget components were presented contralaterally to the target component. However, performance was not impaired when the nontarget components were presented dichotically with an ILD of 20 dB, so that they were perceived contralaterally to the target component. It is concluded that the level of performance in the mistuning task is determined by whether the target is presented to the same ear as the rest of the complex, rather than by its perceived location. PMID- 9857511 TI - Behavioral assessment of acoustic parameters relevant to signal recognition and preference in a vocal fish. AB - Acoustic signal recognition depends on the receiver's processing of the physical attributes of a sound. This study takes advantage of the simple communication sounds produced by plainfin midshipman fish to examine effects of signal variation on call recognition and preference. Nesting male midshipman generate both long duration (> 1 min) sinusoidal-like "hums" and short duration "grunts." The hums of neighboring males often overlap, creating beat waveforms. Presentation of humlike, single tone stimuli, but not grunts or noise, elicited robust attraction (phonotaxis) by gravid females. In two-choice tests, females differentiated and chose between acoustic signals that differed in duration, frequency, amplitude, and fine temporal content. Frequency preferences were temperature dependent, in accord with the known temperature dependence of hum fundamental frequency. Concurrent hums were simulated with two-tone beat stimuli, either presented from a single speaker or produced more naturally by interference between adjacent sources. Whereas certain single-source beats reduced stimulus attractiveness, beats which resolved into unmodulated tones at their sources did not affect preference. These results demonstrate that phonotactic assessment of stimulus relevance can be applied in a teleost fish, and that multiple signal parameters can affect receiver response in a vertebrate with relatively simple communication signals. PMID- 9857513 TI - On the source-identification method. AB - The source identification method is a standard psychophysical procedure for studying the ability of listeners to localize the source of a sound. The method can be described in terms of a statistical model in which listeners' responses are determined by the width and bias of an internal distribution. This article presents a theoretical study of the method, particularly the relationships between the average experimental observables, rms error and variability, and parameters of the internal distribution. The theory is tested against source identification experiments, both easy and difficult. Of particular interest is the experimental dependence of observable statistics on the number of sources in the stimulus array, compared with theoretical predictions. It is found that the model gives a good account of several systematic features seen in the experiments. The model leads to guidelines for the design and analysis of source identification experiments. PMID- 9857514 TI - Methods of interval selection, presence of noise and their effects on detectability of repetitions and prolongations. AB - Accurate methods for locating specific types of stuttering events are necessary for diagnosis, treatment, and prognosis. A factor that could add variability to assessment of stuttering is noise on recordings. The effects of noise were assessed by adding noise to intervals of speech containing all fluent material, fluent material with a repetition, or fluent material with a prolongation. These intervals allow a unique dysfluency response to be made. A statistical analysis of the occurrence of such intervals in spontaneous speech showed that only a limited number of intervals met these criteria. This demonstrated that selecting intervals at random from spontaneous speech (as in time interval analysis procedure) will infrequently lead to a unique and unambiguous dysfluency specification for the interval. Intervals were selected for testing from the intervals that met the stipulated criteria. These were presented for dysfluency judgment when the position of the stuttering within an interval was varied and with different amounts of added noise (no added noise, 3 dB, and 6 dB of noise relative to mean speech amplitude). Accuracy in detecting stuttering type depended on noise level and the stuttering's position in the interval, both of which also depended on the type of stuttering: Noise level affected detection of repetitions more than prolongations: Repetitions were more difficult to detect when they occurred at the end of an interval whereas prolongations were more difficult to detect when they were at the beginning of an interval. The findings underline the importance of adopting rigorous recording standards when speech is to be employed to make stuttering assessments. PMID- 9857515 TI - Role of experience for language-specific functional mappings of vowel sounds. AB - Studies involving human infants and monkeys suggest that experience plays a critical role in modifying how subjects respond to vowel sounds between and within phonemic classes. Experiments with human listeners were conducted to establish appropriate stimulus materials. Then, eight European starlings (Sturnus vulgaris) were trained to respond differentially to vowel tokens drawn from stylized distributions for the English vowels /i/ and /I/, or from two distributions of vowel sounds that were orthogonal in the F1-F2 plane. Following training, starlings' responses generalized with facility to novel stimuli drawn from these distributions. Responses could be predicted well on the bases of frequencies of the first two formants and distributional characteristics of experienced vowel sounds with a graded structure about the central "prototypical" vowel of the training distributions. Starling responses corresponded closely to adult human judgments of "goodness" for English vowel sounds. Finally, a simple linear association network model trained with vowels drawn from the avian training set provided a good account for the data. Findings suggest that little more than sensitivity to statistical regularities of language input (probability density distributions) together with organizational processes that serve to enhance distinctiveness may accommodate much of what is known about the functional equivalence of vowel sounds. PMID- 9857516 TI - The recognition of sentences in noise by normal-hearing listeners using simulations of cochlear-implant signal processors with 6-20 channels. AB - Sentences were processed through simulations of cochlear-implant signal processors with 6, 8, 12, 16, and 20 channels and were presented to normal hearing listeners at +2 db S/N and at -2 db S/N. The signal-processing operations included bandpass filtering, rectification, and smoothing of the signal in each band, estimation of the rms energy of the signal in each band (computed every 4 ms), and generation of sinusoids with frequencies equal to the center frequencies of the bands and amplitudes equal to the rms levels in each band. The sinusoids were summed and presented to listeners for identification. At issue was the number of channels necessary to reach maximum performance on tests of sentence understanding. At +2 dB S/N, the performance maximum was reached with 12 channels of stimulation. At -2 dB S/N, the performance maximum was reached with 20 channels of stimulation. These results, in combination with the outcome that in quiet, asymptotic performance is reached with five channels of stimulation, demonstrate that more channels are needed in noise than in quiet to reach a high level of sentence understanding and that, as the S/N becomes poorer, more channels are needed to achieve a given level of performance. PMID- 9857517 TI - Effects of noise and spectral resolution on vowel and consonant recognition: acoustic and electric hearing. AB - Current multichannel cochlear implant devices provide high levels of speech performance in quiet. However, performance deteriorates rapidly with increasing levels of background noise. The goal of this study was to investigate whether the noise susceptibility of cochlear implant users is primarily due to the loss of fine spectral information. Recognition of vowels and consonants was measured as a function of signal-to-noise ratio in four normal-hearing listeners in conditions simulating cochlear implants with both CIS and SPEAK-like strategies. Six conditions were evaluated: 3-, 4-, 8-, and 16-band processors (CIS-like), a 6/20 band processor (SPEAK-like), and unprocessed speech. Recognition scores for vowels and consonants decreased as the S/N level worsened in all conditions, as expected. Phoneme recognition threshold (PRT) was defined as the S/N at which the recognition score fell to 50% of its level in quiet. The unprocessed speech had the best PRT, which worsened as the number of bands decreased. Recognition of vowels and consonants was further measured in three Nucleus-22 cochlear implant users using either their normal SPEAK speech processor or a custom processor with a four-channel CIS strategy. The best cochlear implant user showed similar performance with the CIS strategy in quiet and in noise to that of normal-hearing listeners when listening to correspondingly spectrally degraded speech. These findings suggest that the noise susceptibility of cochlear implant users is at least partly due to the loss of spectral resolution. Efforts to improve the effective number of spectral information channels should improve implant performance in noise. PMID- 9857518 TI - The relation between identification and discrimination of vowels in young and elderly listeners. AB - This study examined both the identification and discrimination of vowels by three listener groups: elderly hearing-impaired, elderly normal-hearing, and young normal-hearing. Each hearing-impaired listener had a longstanding symmetrical, sloping, mild-to-moderate sensorineural hearing loss. Two signal levels [70 and 95 dB sound-pressure level (SPL)] were selected to assess the effects of audibility on both tasks. The stimuli were four vowels, /I,e, epsilon, ae/, synthesized for a female talker. Difference limens (DLs) were estimated for both F1 and F2 formants using adaptive tracking. Discrimination DLs for F1 formants were the same across groups and levels. Discrimination DLs for F2 showed that the best formant resolution was for the young normal-hearing group, the poorest was for the elderly normal-hearing group, and resolution for the elderly hearing impaired group fell in between the other two at both signal levels. Only the elderly hearing-impaired group had DLs that were significantly poorer than those of the young listeners at the lower, 70 dB, level. In the identification task at both levels, young normal-hearing listeners demonstrated near-perfect performance (M = 95%), while both elderly groups were similar to one another and demonstrated lower performance (M = 71%). The results were examined using correlational analysis of the performance of the hearing-impaired subjects relative to that of the normal-hearing groups. The results suggest that both age and hearing impairment contribute to decreased vowel perception performance in elderly hearing-impaired persons. PMID- 9857519 TI - Long-range acoustic detection and localization of blue whale calls in the northeast Pacific Ocean. AB - Analysis of acoustic signals recorded from the U.S. Navy's SOund SUrveillance System (SOSUS) was used to detect and locate blue whale (Balaenoptera musculus) calls offshore in the northeast Pacific. The long, low-frequency components of these calls are characteristic of calls recorded in the presence of blue whales elsewhere in the world. Mean values for frequency and time characteristics from field-recorded blue whale calls were used to develop a simple matched filter for detecting such calls in noisy time series. The matched filter was applied to signals from three different SOSUS arrays off the coast of the Pacific Northwest to detect and associate individual calls from the same animal on the different arrays. A U.S. Navy maritime patrol aircraft was directed to an area where blue whale calls had been detected on SOSUS using these methods, and the presence of vocalizing blue whale was confirmed at the site with field recordings from sonobuoys. PMID- 9857520 TI - The neural network classification of false killer whale (Pseudorca crassidens) vocalizations. AB - This study reports the use of unsupervised, self-organizing neural network to categorize the repertoire of false killer whale vocalizations. Self-organizing networks are capable of detecting patterns in their input and partitioning those patterns into categories without requiring that the number or types of categories be predefined. The inputs for the neural networks were two-dimensional characterization of false killer whale vocalization, where each vocalization was characterized by a sequence of short-time measurements of duty cycle and peak frequency. The first neural network used competitive learning, where units in a competitive layer distributed themselves to recognize frequently presented input vectors. This network resulted in classes representing typical patterns in the vocalizations. The second network was a Kohonen feature map which organized the outputs topologically, providing a graphical organization of pattern relationships. The networks performed well as measured by (1) the average correlation between the input vectors and the weight vectors for each category, and (2) the ability of the networks to classify novel vocalizations. The techniques used in this study could easily be applied to other species and facilitate the development of objective, comprehensive repertoire models. PMID- 9857521 TI - The effect of abdominal wall morphology on ultrasonic pulse distortion. Part I. Measurements. AB - The relative importance of the fat and muscle layers of the human abdominal wall in producing ultrasonic wavefront distortion was assessed by means of direct measurements. Specimens employed included six whole abdominal wall specimens and twelve partial specimens obtained by dividing each whole specimen into a fat and a muscle layer. In the measurement technique employed, a hemispheric transducer transmitted a 3.75-MHz ultrasonic pulse through a tissue section. The received wavefront was measured by a linear array translated in the elevation direction to synthesize a two-dimensional aperture. Insertion loss was also measured at various locations on each specimen. Differences in arrival time and energy level between the measured waveforms and computed references that account for geometric delay and spreading were calculated. After correction for the effects of geometry, the received waveforms were synthetically focused. The characteristics of the distortion produced by each specimen and the quality of the resulting focus were analyzed and compared. The measurements show that muscle produces greater arrival time distortion than fat while fat produces greater energy level distortion than muscle, but that the distortion produced by the entire abdominal wall is not equivalent to a simple combination of distortion effects produced by the layers. The results also indicate that both fat and muscle layers contribute significantly to the distortion of ultrasonic beams by the abdominal wall. However, the spatial characteristics of the distortion produced by fat and muscle layers differ substantially. Distortion produced by muscle layers, as well as focal images aberrated by muscle layers, show considerable anisotropy associated with muscle fiber orientation. Distortion produced by fat layers shows smaller scale, granular structure associated with scattering from the septa surrounding individual fat lobules. Thick layers of fat may be expected to cause poor image quality due to both scattering and bulk absorption effects, while thick muscle layers may be expected to cause focus aberration due to large arrival time fluctuations. Correction of aberrated focuses using time-shift compensation shows more complete correction for muscle sections than for fat sections, so that correction methods based on phase screen models may be more appropriate for muscle layers than for fat layers. PMID- 9857522 TI - The effect of abdominal wall morphology on ultrasonic pulse distortion. Part II. Simulations. AB - Wavefront propagation through the abdominal wall was simulated using a finite difference time-domain implementation of the linearized wave propagation equations for a lossless, inhomogeneous, two-dimensional fluid as well as a simplified straight-ray model for a two-dimensional absorbing medium. Scanned images of six human abdominal wall cross sections provided the data for the propagation media in the simulations. The images were mapped into regions of fat, muscle, and connective tissue, each of which was assigned uniform sound speed, density, and absorption values. Propagation was simulated through each whole specimen as well as through each fat layer and muscle layer individually. Wavefronts computed by the finite-difference method contained arrival time, energy level, and wave shape distortion similar to that in measurements. Straight ray simulations produced arrival time fluctuations similar to measurements but produced much smaller energy level fluctuations. These simulations confirm that both fat and muscle produce significant wavefront distortion and that distortion produced by fat sections differs from that produced by muscle sections. Spatial correlation of distortion with tissue composition suggests that most major arrival time fluctuations are caused by propagation through large-scale inhomogeneities such as fatty regions within muscle layers, while most amplitude and waveform variations are the result of scattering from smaller inhomogeneities such as septa within the subcutaneous fat. Additional finite-difference simulations performed using uniform-layer models of the abdominal wall indicate that wavefront distortion is primarily caused by tissue structures and inhomogeneities rather than by refraction at layer interfaces or by variations in layer thicknesses. PMID- 9857523 TI - In vitro characterization of a novel, tissue-targeted ultrasonic contrast system with acoustic microscopy. AB - Targeted ultrasonic contrast systems are designed to enhance the reflectivity of selected tissues in vivo [Lanza et al., Circulation 94, 3334 (1996)]. In particular, these agents hold promise for the minimally invasive diagnosis and treatment of a wide array of pathologies, most notably tumors, thromboses, and inflamed tissues. In the present study, acoustic microscopy was used to assess the efficacy of a novel, perfluorocarbon based contrast agent to enhance the inherent acoustic reflectivity of biological and synthetic substrates. Data from these experiments were used to postulate a simple model describing the observed enhancements. Frequency averaged reflectivity (30-55 MHz) was shown to increase 7.0 +/- 1.1 dB for nitrocellulose membranes with targeted contrast. Enhancements of 36.0 +/- 2.3 dB and 8.5 +/- 0.9 dB for plasma and whole blood clots, respectively, were measured between 20 and 35 MHz. A proposed acoustic transmission line model predicted the targeted contrast system would increase the acoustic reflectivity of the nitrocellulose membrane, whole blood clot, and fibrin plasma clot by 2.6, 8.0, and 31.8 dB, respectively. These predictions were in reasonable agreement with the experimental results of this paper. In conclusion, acoustic microscopy provides a rapid and sensitive approach for in vitro chracterization, development, and testing of mathematical models of targeted contrast systems. Given the current demand for targeted contrast systems for medical diagnostic and therapeutic use, the use of acoustic microscopy may provide a useful tool in the development of these agents. PMID- 9857524 TI - Does hearing aid benefit increase over time? AB - In a recent tutorial for the journal, Palmer et al. [J. Acoust. Soc. Am. 103, 1705-1721 (1998)] reviewed the literature on the potential for increases in hearing aid benefit over time (acclimatization). Their review might leave some readers with the impression that acclimatization has implications for the fitting and selection of hearing aids today. We (Turner and Bentler), along with two other researchers in the field (Humes and Cox), conducted a similar review of the literature a few years earlier [Turner et al., Ear and Hearing 17, 14S-28S (1996)] and found little evidence of a robust effect. The bulk of the existing evidence, including the most recent studies on this topic, support earlier conclusions, i.e., that there is no evidence for the existence of a strong acclimatization effect in current hearing aid use. PMID- 9857525 TI - Use of image-based information in judgments of surface-reflectance properties. AB - We examined how well we can recover surface-reflectance properties from shading patterns under changes in surface shape. The stimulus we used was a square surface modulated in depth by a low-pass-filtered random field and rendered by the Phong illumination model [Commun. ACM 18, 311 (1975)]. Two different surface images (target and match) were presented side by side, with either the viewing direction or the surface-normal direction rotating around the horizontal axis. The target shape was manipulated by changing the spatial spectrum, and the target reflectance was manipulated by changing the diffuse-reflection coefficient and the specular-reflection exponent (shininess) of the Phong model. The shape parameters of the match stimulus were fixed, but its reflectance parameters were under the control of subjects, who had to make the apparent reflectance of the two surfaces as similar as possible. The results showed that the constant error (difference between simulated and matched values) was large except when the two surfaces had the same shape parameters or when they differed only in scale. The pattern of the constant errors and response variabilities suggests that the judgments of the subjects were based on the similarity of the luminance histogram of the surface image. Our results demonstrate a limitation of surface-reflectance constancy for changes in shape and the importance of image-based information in reflectance judgments. The results are discussed in relation to previous studies that showed effects of spatial layout on surface-reflectance perception. PMID- 9857526 TI - Staining tissue with methacrylate casts for light microscopy. AB - Scanning electron microscopy (SEM) of methyl methacrylate casts and light microscopy (LM) of tissue are well-established methods for studying the microcirculation. The two are complimentary, but methacrylate is transparent and thus its presence is often not appreciated by LM. Applying histologic stains to sections of tissue embedded in methyl methacrylate would allow the relationships of light microscopic and scanning electron microscopic views of cast vasculature to be better appreciated. We sought to test different stains on cast tissue to find one that would accent the cast. Surgically removed and autopsied human lungs were cast with methacrylate and processed by routine light microscopic methods. They were stained with the hematoxylin and eosin, Masson trichome, elastic--van Gieson, Grocott methenamine silver, Brown-Brennan, and Ziehl-Neelsen methods. The Ziehl-Neelsen procedure stained the methacrylate best, giving it a red color. This procedure also worked well without heating. We conclude that (1) cast methacrylate lung can be processed for routine LM with excellent results; (2) methacrylate stains well with the Ziehl-Neelsen technique; (3) the acid--fast stained cast lung shows capillaries and cells in both normal and diseased lung better than the routine hematoxylin and eosin stain; (4) this technique can be used to assess filling and correlate findings on the same tissue with the two different microscopic methods. PMID- 9857527 TI - Confocal laser scanning microscopy of chondrocytes in vitro: cytoskeletal changes after quinolone treatment. AB - The use of quinolone antibiotics would be significant for chronically diseased children (e.g., cystic fibrosis) as a prophylactic long-term treatment. However, quinolones were shown to cause cartilage damage in experimental animals when administered during certain developmental stages. In the present study, the effect of quinolones on chondrocytes was studied in a cell culture model in order to avoid animal experiments, to investigate the influence of single factors, and to open up the possibility to test human tissue. Chondrocytes were obtained from hip joint cartilage of 3 to 4-weeks-old rats and cultured in control medium or quinolone-supplemented medium. It was shown that quinolones heavily disturbed adhesion of chondrocytes to the culture dish, accompanied by changes in cell shape and cytoskeletal morphology. Reduction of filamentous actin (stress fibers) and disintegration of vimentin fibers was demonstrated by immunofluorescence and evaluated by confocal laser scanning microscopy. In contrast, distribution and amount of the adhesion molecule integrin alpha 1 did not change. Results of the present study indicate that quinolones disturb the adherence mechanism of chondrocytes and lead to cytoskeleton changes. PMID- 9857528 TI - Errors in quantitative backscattered electron analysis of bone standardized by energy-dispersive x-ray spectrometry. AB - Backscattered electron (BSE) imaging has proven to be a useful method for analyzing the mineral distribution in microscopic regions of bone. However, an accepted method of standardization has not been developed, limiting the utility of BSE imaging for truly quantitative analysis. Previous work has suggested that BSE images can be standardized by energy-dispersive x-ray spectrometry (EDX). Unfortunately, EDX-standardized BSE images tend to underestimate the mineral content of bone when compared with traditional ash measurements. The goal of this study is to investigate the nature of the deficit between EDX-standardized BSE images and ash measurements. A series of analytical standards, ashed bone specimens, and unembedded bone specimens were investigated to determine the source of the deficit previously reported. The primary source of error was found to be inaccurate ZAF corrections to account for the organic phase of the bone matrix. Conductive coatings, methylmethacrylate embedding media, and minor elemental constituents in bone mineral introduced negligible errors. It is suggested that the errors would remain constant and an empirical correction could be used to account for the deficit. However, extensive preliminary testing of the analysis equipment is essential. PMID- 9857529 TI - [Phenomenon of karyolysis in cytophotomorphometric examinations]. AB - Studies were performed on hepatocytes of ten rabbits' livers. The one-year-old animals were sacrificed by a blow in the suboccipital region. Under aseptic conditions each liver was divided into 9 parts. One of them was examined at once (zero time) the remaining parts were placed in sterile vessels lined with gauze moistened in Ringer's solution, which created an environment of 100% humidity. Autolysis was being conducted for 8 hours at temperature 37 degrees C, carrying out the investigations with hourly intervals. The cells for studying were obtained by preparing contact smears that stemmed from a fresh cross-section of the liver segment. The smears were fixed in formalin vapour 40% proceeding for 2 hours at temperature 21 degrees C, and subsequently in 96% ethanol for 0.5 hour. Feulgen's reaction was run in accordance with the routine principles by using the basic fuschin NI 42510 of Fischer Firm (USA). The morpho-photometric measurements of nuclei were taken by means of computed device "Morphoquant" of Carl Zeiss Firm, Jena, 1982. The following measurements of the cell nuclei were accomplished, namely: A. morphometric--surface, circumferential length, bulged surface, ratio of diameters, unstained surface; B. photometric--medium extinction, circumferential extinction, total extinction. The results of studies were documented by 95 diagrams. On the basis of measurements taken, it has been established that, during the 7 hour-long period of autolysis, nucleus of hepatocyte is subjected to dynamic transformations, whereas the plotted function does not follow a linear course. During the observation the circumference and the surface resemble a polynomial with three extremes (Fig. 12, 21, 39); similarly the mean extinction and circumference extinction (attention is attracted by its severe course as compared to surface and circumference) (Fig. 57, 66). At the time of experiment the differentiation of nuclei population undergoes changes in the direction of its uniformity which concerns, to greater or lesser degree, all the investigated features (compare figures of histograms). The isolated group of tetraploidal nuclei is characterised by occupying another place. It is defined by 3 variables in spatial arrangement (Fig. 76, 80). The analysis of correlation allowed for determination of the dynamic connections, between the studied variables, which seem to be specific for a given experimental time (Tab. 1, 2). On the ground of the performed studies 7 final conclusions have been formulated: 1. Autolysis of the nuclei of hepatocytes is non-linear phenomenon; (its course resembles multinominal graph 6 degrees with three extremes in 1, 4 and 6th hour. 2. Photomorphometric parameters allow for determining the status of the nucleus of hepatocyte in the course of autolysis. 3. The fourth hour of autolysis is a characteristic point after which no response of nucleus to deteriorating oxygen condition was observed. 4. In postmortem autolysis of the nuclei of hepatocytes pyknosis is found to appear, there is no phenomenon of karyorrhexis. 5. After a 7 hour-long experiment, Feulgen's diffusional reaction starts in cytoplasm. 6. A group of tetraploid nuclei is being isolated in the course of the autolysis of hepatocytes. 7. In the course of experiment there appeared a small quantitative loss of nuclear DNA. PMID- 9857530 TI - [Determination of the usefulness of selected biochemical parameters for assessing the advanced atheromatous changes in human coronary arteries]. AB - It is generally accepted that atherosclerosis is a dynamic process in which many factors of lipid, hemostatic or other nature play their negative and positive roles. The purpose of the study was to determine the relationship between the atheromatous changes in coronary arteries being assessed angiographically and the lipid and hemostatic risk factors, as well as to select biochemical parameters, which would be helpful for prognosing the degree of intensity with regard to atheromatous changes in coronary arteries. Studies of lipid parameters and hemostasis system were performed in 31 men with atherosclerosis of coronary vessels being angiographically estimated. The degree of intensity concerning the atheromatous changes was defined in a point scale according to Gensini based on the magnitude of coronary artery stenosis and its localization in respect of significance for myocardial function. The studied patients were divided into two groups, which differed by the degree of the intensity of atheromatous changes in coronary arteries: group I--men with mild (M-CAD, score < 32) n = 15, group II- men with severe atherosclerotic changes (S-CAD, score > or = 32) n = 16. The characteristics of both groups are given in table 1. All patients were on nitrates, salicylates, beta-blockers and calcium channel blockers. No antilipemics or anticoagulants were administered. The following biochemical parameters were determined in all men: cholesterol-Ch; triglycerides-TG; phospholipids-PL; apolipoproteins: Apo A, Apo A-I, Apo B; lipoproteins: VLDL, LDL, HDL and their lipids and proteins components; lipoprotein (a)-Lp(a); fibrinogen-Fb; euglobulin lysis time-ELT; inhibitor tissue plasminogen activator PAI-1; antithrombin III--AT III; spontaneous platelet aggregation-SPA, platelet factor 4-PF 4 and glucose. Table 2 lists the lipid parameters in serum and lipoprotein fractions. The levels for apolipoproteins A, A-I, B, lipoprotein (a), hemostatic parameters and glucose are given in table 3. Tables 4 and 5 present the results of multiple regression analysis for severity of atherosclerotic changes (score--dependent variable y) lipid and hemostatic parameters and glucose (independent variables x) in both groups. Prognostic variables necessary for the best fit in the model of relationship studied have been selected. Independent variables x are listed in descending order according to the absolute value of b*x. On the basis of the performed statistical analysis of the results of studies it has been ascertained that the biochemical parameters differentiating the patients with regard to the intensity of atheromatous changes are the coefficients: LDL-Ch/HDL-Ch and Apo B/Apo A ratio, LDL-PL, Fb and ELT whose values were higher as well as HDL-Apo A-I whose value was lower in the group of men with more severe atherosclerotic changes in coronary arteries (S-CAD). The stepwise multivariate analysis indicates that the most profound prognostic significance in risk of coronary atherosclerosis is claimed successively by: glucose, LDL-PL, HDL-Apo A-I, AT III, Fb, ELT, PAI-1, SPA, Lp(a), Apo B and PF 4. The results of the accomplished studies point out that the above-mentioned lipid, hemostatic parameters and glucose may be helpful in prognosing the severity of coronary atherosclerosis. PMID- 9857531 TI - [An attempt to block histamine release from basophils granulocytes with antibodies obtained as a result of long-term immunization]. AB - Pathogenetic mechanisms responsible for efficacy of specific immunotherapy still remain to be fully explained. This concerns both desensitization with classic allergens and very rarely used specific immunotherapy with bacteria. Microbes can play important role as hypersensitivity factor in some allergo-inflammatory processes. Bacterial products may act as basophil histamine liberators through immunological (IgE-mediated) and nonimmunological--particular lectin-sugar way. The aim of study was to verify if histamine release triggered by microbes could be modified (blocked) with specific antibacterial antibodies--taking into consideration both of mechanisms of basophil degranulation. The size of immediate (in healthy persons--Tab. 3, 4) and late as well as delayed (in asthmatic patients--Tab. 8) skin reactivity to examined microorganisms and the degree of basophil histamine release induced with these bacteria were compared. Human basophils were isolated from peripheral blood on Ficoll-Hypaque gradient, next challenged with whole, formalin-killed bacteria and with the same bacteria after incubation with specific and nonspecific sera. To differentiate between IgE dependent and non-immunological mechanisms of histamine release, the IgE molecules were removed from the surface of the basophils by exposure to pH 3.6 (stripping). In each experiment histamine release induced by anti-IgE antibodies was used as control of stripping (Tab. 5, 9). Levels of histamine from the basophils (without and after stripping) incubated with non-coated and specific antibodies coated bacteria were compared. The results were expressed as a percentage of total histamine content in the sample. Histamine release was assayed spectrofluorometrically by using Shore method in Norn modification. The main investigations concerned the basophils from 12 healthy, non-atopic individuals, who had positive immediate skin reactions with at least 1 from 3 microbial strains: Staphylococcus aureus 9615 (unencapsulated), Staphylococcus aureus Smith (encapsulated) and Escherichia coli. Sera containing specific antibodies for these microorganisms were obtained from immunized rabbits. As negative control served sera collected from animals after immunization. Additionally the basophils of 6 asthmatic (intrinsic asthma) patients treated with autovaccines were examined. All patients demonstrated positive late and delayed skin reactions, 3 of them also immediate, to autologous Neisseria and Moraxella species cultured from upper respiratory tract. The bacteria were used as a component of autovaccine and as a basophils stimulating factor in histamine assay. Microbes were incubated with patients own sera before (unspecific serum) and after treatment (source of "specific" antibodies). CONCLUSIONS: 1. Bacteria induced basophil histamine release through two ways: immunological (IgE-mediated) and non-immunological (sugar-lectin interactions). 2. Non-immunological interactions played the main role in basophil histamine release induced by bacteria--both in normal individuals and asthmatic patients. 3. Sera of immunized with bacteria animals partially reduced basophil histamine release induced by homologous strains (Tab. 7). 4. An incubation of autologous bacterial strains with asthmatic patients's sera collected after autovaccines treatment has no influence on basophil histamine release induced by these microbes (Tab. 9). 5. There was no correlation between the skin reactivity to bacteria (both in healthy persons and in asthmatic patients) and the intensity of basophil histamine release induced by microbes. PMID- 9857532 TI - [Skin reactions to antigens of propionibacterium acnes in patients with acne vulgaris treated with autovaccine]. AB - One of the most common diseases of the skin is acne. The etiology and pathogenesis of acne, in spite of the advancement of medical knowledge, remain unknown and the effects of treatment unsatisfactory. The mechanism of the beneficial effects of immunotherapy in some cases of acne, including autovaccines prepared from the bacterial strains of the patient, also awaits explanation. The present work was aimed at elucidating the influence of autovaccine on some parameters of specific humoral and cellular response against the same strains of Propionibacterium acnes that were isolated from the patient to prepare the autovaccine. These parameters were evaluated in vivo on the basis of early and delayed skin reactions and in vitro using respective laboratory tests. By analysing the influence of autovaccine on the immunological status it was hoped to shed some light on the immunological aspects of acne. An improvement after autovaccine was noted in 47.6% of patients. At the same time it was observed that the results of the present treatment with autovaccine were much better in patients who were previously treated for acne with Acnevac or autovaccine than in other patients. One may therefore conclude that repeated immunotherapy in acne is advantageous in terms of results. The frequency of early skin reactions against the Propionibacterium acnes of the patient and against standard strains, the level of serum IgE antibodies in patients with acne of various intensity and the release of histamine in the presence of Propionibacterium acnes from basophils of patients with positive early skin reactions all stand against the role of early type hypersensitivity and anaphilactoid phenomena linked with the structural antigens of the patient's strains of Propionibacterium acnes in the pathogenesis of acne (Tab. 1, Fig. 1). Reactions reflecting delayed-type hypersensitivity against the patient's strains of Propionibacterium acnes were observed more frequently than early-type reactions and more frequently than against standard strains (Tab. 2). Clinical improvement was particularly evident in patients in whom the intensity of the reactions decreased after treatment. This was accompanied by higher titres of specific antibodies against structural antigens of Propionibacterium acnes (Tab. 3) and a greater inhibition of migration of mononuclear cells in the presence of these bacteria or a nonspecific mitogen (PHA). It was concluded that specific antibodies generated by the autovaccine and directed against the strain of Propionibacterium acnes of the patient may reduce the intensity of delayed-type reactions in some cases of acne, as previously reported for tularemia and tuberculosis. Final unequivocal conclusions as to the pathogenesis of acne and mechanism of the effects of autovaccine could not be drawn. However, the present results form an encouraging basis for further research in this field. PMID- 9857533 TI - [Analysis of factors influencing the results of surgical treatment of esophageal cancer based on personal material]. AB - The aim of the study was to evaluate some factors which could influence postoperative course as well as early and late results of surgical treatment of thoracic esophageal cancer. 75 out of 90 patients treated surgically in the Department of General and Transplantation Surgery, Pomeranian Medical Academy, Szczecin were involved in the investigation. Basic laboratory tests, X-ray examination, endoscopy, tissue diagnosis, pulmonary ventilation study (since 1991) and ECG were done in all of them. The analyzed group of patients consisted of 71 males and 4 females. The period of follow-up ranged from 6 months to 10 years. The mean age of patients was 56.1 years and duration of dysphagia 4.62 months. The patients of stage II disease predominated and those in stage I comprised 12%. The weight loss was 8.89 kilograms and length of lesion 6.53 centimeters on average. Squamous cell carcinoma group was the largest and consisted of 63 patients, 7 of them had preoperative radiation therapy, 54 patients were operated on by right transthoracic approach following previous laparotomy. Esophagectomy was done by the method of Orringer, Akiyama and through the left transthoracic approach (following laparotomy) in 8, 2 and 3 patients respectively. In 8 patients esophagectomy was performed by the left transthoracic approach only. The operation was considered radical in 42 percent of patients. The most frequent tumor location was the mid esophagus appearing in 44 patients; 28 patients underwent chemotherapy postoperatively. Postoperative complications were found in 88 percent of patients. All the patients were divided into groups depending on the analyzed factor. The following factors have been investigated: age of patients, duration of dysphagia, stage of disease with a separate evaluation of T and N factors, weight loss, length of lesion, tissue diagnosis, preoperative radiation therapy, way of resection, tumor location, postoperative chemotherapy, effect of department and operating surgeon, complications postoperatively. It was found that both early and late results depended significantly on stage of disease including "T" and "N" factors (Tab. 2, 3, 4), tissue diagnosis (Tab. 5), type of resection (Tab. 6), location of tumor (Tab. 8), postoperative chemotherapy (Tab. 9) and experience of the department. Stage of disease together with method of operation have significantly influenced the incidence of cardiovascular complications encountered (Tab. 1, 2). Some topics of importance connected with the presented factors were dealt with in discussion, and studies available in literature regarding the outcome of modern directions concerning the treatment of esophageal cancer were also presented. PMID- 9857534 TI - [Evaluation of factors influencing the result of surgical treatment in patients with ruptured aneurysm of the abdominal aorta and iliac arteries]. AB - The aim of this paper was to analyze the risk factors influencing early and late results of postoperative therapy in patients with ruptured abdominal aortic aneurysm. The analysis included: age of patients, type and duration of symptoms, presence of shock prior to surgery, size of aneurysm, place of rupture, type of surgery and prosthesis, amount of blood transfused during the surgery, duration of the operation and occlusion. In conclusion the early results were evaluated on the basis of postoperative mortality. The examinations involved a group of 56 patients (49 males, 7 females) who had urgent surgeries in the years 1988-1995. Evaluation of the early results were based on case histories, results of some tests, anaesthetic charts and ICU files. The mean age was 66.7. During the surgery 7 patients died due to severe hemorrhagic shock. In the postoperative period 20 patients died due to complications. As a result the dependence between survival and 1) the presence of severe posthemorrhagic shock (Tab. 1), 2) the diseases the patients have suffered from particularly those of circulatory system (Tab. 4), 3) the type of surgery and prosthesis (Tab. 2, 3), 4) complications of postoperative period have been confirmed (Tab. 5). Other factors did not have any significant influence on the surgery. Examinations of the late results in 29 patients (the observation lasted 6-20 months) have been carried out. From this group--22 patients reported for follow-up examinations, 5--died, 1--lives abroad. The follow-up examinations revealed coexistence of hypertension in more than 60% of the patients and obliterative changes of peripheral arteries in about 30%. USG examination of the implanted prosthesis showed a proper blood flow in all patients, the presence of aneurysm of pseudoanastomosis has not been found. PMID- 9857535 TI - [The influence of cumulative dexamethasone, promethazine and dextran 70 used as protection against intraperitoneal adhesions on selected parameters of humoral immunity in women operated on for infertility]. AB - One of the most frequent reasons of the intraperitoneal adhesions formation in young women is pelvic inflammatory disease and surgical operations of pelvic organs. The element of more complex activities against adhesion creation is to limit an inflammatory reaction in the area of operation by intraperitoneal and systemic pharmacotherapy. The system of activities put into practice in the Clinic of Gynaecology includes in the field of pharmacological prophylaxis a systemic and intraperitoneal use of dexamethasone and promethazine before, during and after the operation, in total doses respectively: 250 mg and 300 mg, as well as intraperitoneal dosing of 500 ml of dextran 70, just before closing the peritoneal cavity. Verification of the results of these activities during laparoscopy or laparotomy shows its high effectiveness. For many years we did not observe any complications of any wound healing after the operations. Due to existing reports about possible complications in the postoperation period, which are probably caused by the use of the above-mentioned pharmaceutics, I decided to investigate how dexamethasone, promethazine and dextran 70 influence the selected parameters of humoral resistance in women who were operated on due to mechanical infertility. The investigated group consisted of 30 women, while the control group comprised 10 women who had gynaecological operations of similar extent. The venal blood samples were collected before the operation and on the third, fifth and twenty first day after the operation. The levels of selected acute phase proteins were checked and the character as well as the dynamics of value changes were monitored in both groups. Because of great variability between the proteins the fractions were picked out focusing on the differences in dynamics of metabolism, half-life time and biological functions. Following proteins were investigated: C-reactive protein, alfa1-acid glycoprotein, haptoglobin, alfa1 antitripsin, ceruloplazmin, alpha2-macroglobin, hemopexin, immunoglobulins A, G, M, albumins, transferrin, alpha2-HS-glycoprotein. The analysis of the study results has indicated that the application of dexamethasone, promethazine and dextran 70 only exerts modulating influence on the course of acute phase reaction induced by the operations. In the group having been studied earlier (in 3rd postoperative 24-hours) significant changes were revealed concerning the level of particular proteins in accordance with their physiological character (increase or decrease of level) in the control group such changes appeared later (about 5th 24 hours) and were characterized by markedly smaller amplitude and dynamics (Tab. 1 2, Fig. 1-15). The levels of proteins defined prior to operation were reached faster by patients of the study group (up to 21st 24-hours after operation). It has been disclosed that the use of composed antiadhesion therapy with dexamethasone, promethazine and dextran 70 accelerates, intensifies and simultaneously restricts the duration of transformations defined as acute phase reaction, which result in reducing the number of intraperitoneal adhesions. This therapy does not impair the mechanisms of humoral immunity in patients operated on. PMID- 9857536 TI - [Selected parameters of blood platelet function in patients with myeloproliferative syndrome]. AB - Symptoms of hemorrhagic diathesis and/or thrombotic complications are observed in patients with myeloproliferative syndrome (m-p.s.). It is emphasized that there is a meaningful direct and indirect share of blood platelets in the mentioned disturbances of hemostasis. Therefore in own investigations an attempt was made to evaluate the morphology and function of blood platelets in patients with various forms of m-p.s., as well as to find an answer to the question: do any differences in morphology and function (activation) of blood platelets appear in patients with m-p.s.?, and is activation of blood platelets always accompanied by the symptom of hemorrhagic diathesis or past thrombotic episodes (the second ones evidenced in the patients anamneses)?, do the blood platelets in idiopathic thrombocytopenia (one of the forms of m-p.s.), differ morphologically and functionally from platelets in other forms of m-p.s. The investigation comprised 45 patients, in whom m-p.s. was diagnosed at Department of Hematology PAM (28 women, 17 men) aged from 36 to 82 years (mean age 58.10 + 10.61). The diagnosis of m-p.s. in each patient was established on the basis of clinical picture and laboratory examinations (blood morphology, hematocrit, FAG score, histopathological examination of bone marrow). In patients the accomplished determination concerned blood platelets count, mean blood platelet volume, platelocrit, volume range of blood platelets, coagulation time by Duke's method. The function of platelets was estimated with the help of the following studies: aggregation of blood platelets under the influence of ADP according to Born's method, concentration of beta-thromboglobulin (beta-TG) and platelet factor--4 (PF-4) in plasma by ELISA method. In order to pay due consideration to the effect of blood platelets count exerted on the concentration of beta-TG and PF-4, a supplementary parameter was introduced, the so-called beta-TG and PF-4 standardized. That allowed for studying the platelets in all patients under the same conditions. The results of own investigations have shown that most of the patients with m-p.s. had disturbances involving the morphology of blood platelets (Tab. 2) and increased activity of platelets, being expressed by a rise in concentrations of beta-TG and PF-4 in plasma (Fig. 1, 2) and by abnormal aggregation of platelets due to ADP influence. Moreover, the determination of beta-TG and PF-4 concentration in plasma in thrombocytopenia states, particularly in patients with idiopathic thrombocytopenia ought to be performed after the standardization of platelets count, the finding being also supported by the results of own investigations (Tab. 3). There may be compliance with the viewpoint expressed in the literature that the results of studying the morphological parameters of blood platelets and their function (activation) in m p.s. are not a prognostic factor for the appearance of thrombotic complications and/or hemorrhages. PMID- 9857537 TI - [Early and late results of treating trigeminal neuralgia by Gasserian ganglioglycerolysis using a personal subjective pain measurement scale]. AB - The results of treating 61 patients suffering from trigeminal neuralgia by Gasserian ganglioglycerolysis were estimated in the studies. The following features were considered: sex, age, side of pain, duration of symptoms, pharmacological therapy and the course glycerolysis procedure, with attention being paid to whether the CSF outflow is present or not. The ganglionglycerolysis procedure was modified to ensure that the needle is inserted directly in the ganglion, but not into periganglional space. The anesthetic effect was obtained by simultaneous glycerol injection and slow needle withdrawal. The recent and remote results were assessed by implementing own subjective pain sensibility scale, characterizing the therapeutic results and the frequency of disease recurrence. PMID- 9857538 TI - [Horizontal laryngectomy in treatment of laryngeal cancer--oncologic and function results]. AB - The purpose of this paper was the retrospective evaluation of the results after horizontal partial larynx resection of different dimension. The material comprised 468 patients treated according to the described techniques in ENT Clinic Pom. Med. Acad. between 1970-1991. The series of patients subjected to analysis consisted of 35% of all cases treated by surgery and 70% of all conservative operations performed for cancer of the larynx in the above-mentioned period of time. Particulars were obtained from: a) case history, b) control examination, c) correspondence, d) data from USC. The results of the treatment in particular groups were submitted for detailed analysis. The assessment of applied technique from oncologic point of view was done on the basis of local recurrences and percentage of 3 and 5 years survival rate. Functional results were evaluated according to protective, respiratory and phonatory functions of the residual larynx. Results of the investigations were submitted for statistic valuation. In all types of operations satisfactory oncologic results were achieved. They were represented by high percentage of survival rate and low percentage of local recurrences. The longest survival 3- and 5-years respectively (73.90%) and 63.45%) and the lowest index of local failure (4.2%) were seen after classic, supraglottic resection. After horizontal glottic larynx resection 8/10 of patients operated on survived 3 years, 4/6 survived 5 years. Local recurrence was observed in 1/12 patients (Tab. 2). The lowest rate of survival (54.76% and 42.86%) and the highest number of local failures (11.3%) were revented after supracricoid laryngectomy with cricohyoidoepiglottopexy (CHEP)--in this group carcinomas staged T3 were predominant. After enlarged supraglottic laryngectomy and supracricoid resection with cricohyoidopexy (CHP) oncologic results were very close (Fig. 1). No substantial statistical difference was recorded in survival percentage after enlarged supraglottic larynx resection (70.51% and 61.66%) and supracricoid one with CHP (68.54% and 58.43%), nor in percentage of local recurrences (4.4% and 5.7%). The efficiency of the residual larynx was dependent on the extent of the resection. From the discussed types of operations the best functional results were achieved after classic supraglottic and glottic larynx resections. This was distinct in the best quality of speech, efficient respiratory tract protection and high percentage of decannulated cases (93%). In the glottic resection the respiratory tract protection was satisfactory in 9/11 patients and in 7/8 satisfactory speech quality was noticed. Following the extended laryngeal structures resection (extended supraglotic and supracricoid laryngectomies) worse protection of respiratory tract, higher percentage of not decannulated cases and worse quality of speech were observed than in operation previously discussed. It was proved that the one-stage reconstruction of the laryngeal structures (arytenoid cartilage and/or vocal cord) or base of the tongue improves the result considerably, in particular the deglutition and phonation. The oncologic and functional results after the discussed types of operation should stimulate their more frequent application in larynx cancer treatment. PMID- 9857539 TI - [Evaluation of the usefulness of ultrasonic examination of the pre-epiglottic space in patients with laryngeal carcinoma]. AB - The subject of the paper was to assess the usefulness of ultrasound examination of preepiglottic space (Fig. 1) in laryngeal carcinoma. Importance of oncological state of HTE space for the choice of treatment was presented. On the basis of 73 patients having laryngeal cancers with different location and progression (Tab. 1) USG, CT imaging and postoperative histopathological investigation were compared. USG investigation of HTE space was performed with ultrasonograph SSA 250 A (Toshiba) at the frequency of transducer 7.5 MHz. In this paper, the method of USG investigation of preepiglottic space was presented. CT examination of HTE space was performed with GE CT Max-640. On the basis of examinations in a group of 16 healthy persons, characteristics of USG image of normal HTE space was established (Fig. 2). That provided a point of reference involving the assessment of USG image of HTE space in patients with laryngeal cancer (Fig. 3). Results of USG, CT (Fig. 4, Fig. 5) and postoperative histopathological examinations were compared. In 86% of cases agreement between USG and histopathological examinations was noted as was between USG and CT. Results of assessment of USG investigation were: sensitivity 90%, specificity 78%, efficiency 86%. Attention was focused on emphatic predisposition of cancer localized on subhyoid part of epiglottis to penetrate into HTE space. Among the cases with this localization of cancer, in 80% neoplastic infiltration was observed in histopathological investigation of HTE space. In this paper causes of false results were analysed. In USG 6 cases were false positive and 3 false negative. In CT 7 cases were false positive and 2 false negative. Reasons of difficulties in interpretation were: advanced laryngeal carcinoma, short and thick neck, changes in neck spine. Established results have shown that USG investigation of preepiglottic space is valuable method and very useful in preoperative diagnosis in laryngeal cancer. It properly helps to assess oncological state of HTE space. This method makes it possible to find even minor changes in HTE space (0.5 cm). It should be here added that the method is easily applicable, reproducible, and in comparison with CT much cheaper and safe with regard to health. PMID- 9857540 TI - [Evaluation of the diagnostic value of color Doppler ultrasound examination of salivary gland neoplasms and metastatic tumors from the facial bones]. AB - The aim of the study was to evaluate usefulness of colour Doppler ultrasound examination in diagnosing the salivary gland tumours and the metastatic tumours of the neck originating from the facial part of the skull. Epidemiology and histopathology of the neoplasms involving the salivary glands and the facial skeleton were discussed including the route of their spreading to the neck. The author presents update techniques of bony face radiologic imaging and basic principles of modern colour Doppler ultrasound. The examinations with the use of a colour Doppler equipment-Acuson 128-XP 10 were performed in 150 patients with the neck tumours. The exact location, size, morphology and blood supply were assessed using B and B colour mode. Then some big neck vessels like the common, internal and external carotid artery, vertebral artery, internal jugular vein were visualized. All the patients were divided into three groups according to what they were suffering from: sialoadenitis, benign and malignant tumours. The obtained results were compared and confronted with clinical features. The pattern of vascularization failed to allow for establishing preliminary diagnosis in patients in each group. Within the first group, with inflamed glands did not compress the neck vessels. Of all the patients with benign tumours, extrinsic compression on the internal jugular vein and the carotid arteries was found in 16 and 14 patients respectively. In the third group of patients with malignant disease, compression on the veins was detected in 10 cases while 5 tumours compressed the arteries. The invasion involved the internal jugular vein in 7 patients while the common and internal carotid arteries were invaded in 6. The vertebral artery was never found to be affected. It was demonstrated that compression on veins resulted in disturbing the flow which was not observed as far as the arteries were concerned. Disturbing in the flow of veins and arteries was disclosed in cases of invasion. Atheromatosis was detected in patients of each group, 7 of them had to be operated on due to a stricture within the proximal part of internal carotid artery. The results of the study express high diagnostic value of colour Doppler ultrasound examination in the neck tumours. The examination should be obligatory performed as one of the first diagnostic measures after admitting a patient. The method seems to be of very little use in diagnosis differentiating inflammation, benign and malignant salivary gland tumours and metastatic masses of the neck hence some other diagnostic procedures should be here employed. PMID- 9857541 TI - [An attempt to use ultrasonic technique for confirming the diagnosis, planning and observation of long-term treatment results of painful temporo-mandibular joint dysfunction]. AB - The author presents an attempt of using ultrasonographic technique in diagnosis, planning and observation of treatment results of temporo-mandibular joint pain dysfunctions. Temporo-mandibular joint pain dysfunctions are interchangeably also called temporo-mandibular joint functional disorders. The assessment of pain symptoms in temporo-mandibular joint dysfunctions pain symptoms is principally based on a subjective estimation by the examining practitioner. There is no univocal definition of the disease or a simple index evidencing important symptoms in decision making. Additionally X-ray technique examinations, being hitherto used, in early stages of the disorder do not allow to diagnose it, and are also burdensome to a patient. The aim of this study was to confirm visibility of anatomical elements of the temporo-mandibular joint in an ultrasound examination, assess the mobility of the articular disc before, during and after prosthetic treatment with and without the use of ultrasound technique, and to determine the period of time necessary to obtain a therapeutic effect. The study material consisted of 180 patients, 128 women and 52 men, aged 20 to 60 years, treated by applying prostheses because of temporo-mandibular joint pain dysfunction, in the Department of Prosthetic Dentistry of the Pomeranian Medical Academy. The patients were divided into 2 groups, control and study group. The control group consisted of 90 patients, 63 women and 27 men. In this group prosthetic treatment planning and observation of results was based on a subjective estimation of the practitioner. The study group here comprised 90 patients, 65 women and 25 men, aged 26 to 60 years. In this group prosthetic treatment planning and observation of treatment results were carried on with the use of ultrasound technique. Data from both groups concerning history, results of examinations carried out by ultrasound technique, and the assessment of ultrasound examination were noted on standard examination records used in the Department of Prosthetic Dentistry. For the need of this study an own ultrasound technique was elaborated which allowed for analyzing the ultrasound image in a static situation and during functioning. The ultrasound examination was accomplished with the use of Acuson 128 XP apparatus, linear probe 7.5 MHz. In both groups prosthetic treatment was carried out using different prosthetic methods such as: splints, occlusal adjustment on fixed prosthetic restorations in therapeutical occulus et height in abnormal occlusion. In the study group the therapeutical occulus height was determined under the control of ultrasound technique. Statistical analysis of the obtained results was performed with the use of chi-square test and chi-square test with Yates correction. On the basis of the accomplished study it has been determined that the ultrasound technique makes it possible to visualize morphological elements and facilitates functional observation of the temporo-mandibular joint, articular disc, mandibular condyle and lateral pterygoid muscle in a degree which allows for planning the treatment and the observation of results (Tab. 1-6). The use of ultrasound technique in determining spatial position of the mandible in temporo-mandibular joint permitted to objectively estimate examination results in the study group. A small number of publications on the use of ultrasound technique in functional examination of the temporo-mandibular joint accessible in foreign literature and general reference on the subject in Polish literature do not allow for an objective comparison of obtained study results. PMID- 9857542 TI - [Comparative analysis of edentulous patients treated traditionally and with the use of a face-bow and Quick Master articulator]. AB - Correct determination of the occlusal plane is one of the most difficult stages of treatment. After determining the correct occlusal plane its reproduction is possible thanks to the use of articulators. These instruments simulate movements of the jaw in three planes. One of the most optimal articulators is a semi adjustable type. These instruments are not complicated and give good treatment results. A modern semiadjustable type of articulator is Quick Master. The face bow which comes together with this instrument is used for recording and transferring the occlusal relation to the articulator. This allows to mount models in an adequate three dimensional position in relation to the temporo mandibular joint. The use of these instruments leads to many questions and doubts due to difficulties in their use. Therefore the aim of my study was to elaborate a simple method of occlusal recording. I have also compared the treatment results of edentulous patients treated with the use of an articulator and the use of a traditional method. Prosthetic restorations were prepared among 60 patients. The study material was divided into two groups of 30 patients each. In the control group for preparing complete dentures the Gysi method was employed as the most common. In the study group a face-bow and articulator were used. After preparing complete dentures detailed clinical control examinations were carried out and were repeated 24-48 hours after fitting the dentures and also after 3 and 6 months of their use. Working with the face-bow I have employed my own modification of recording the occlusion. The upper wax rim was placed on a slightly warmed bite fork and drawing pins were placed in the recording block to act as a type of key. The lower rim was warmed and brought to occlusal contact a couple of times. Next the face-bow was inserted. The recorded occlusion was transferred and mounted in the articulator. Teeth in both cases were set up similarly to the Gysi method. Lower teeth were set up on the top of the ridge, the upper teeth could be set slightly out of the top line but in the area marked afore between the lines. After preparing and fitting the dentures a clinical examination was carried out and a survey concerning the dentures in use was filled out by the patients. In the study group QM the adaptation period lasted 5 to 30 days, an average of 10.5 days. Full adaptation was achieved in 24 patients during 3 to 14 days. In the control group adaptation period lasted 3 to 42 days, an average of 18.4 days (Tab. 1). A significant difference was obtained. An analysis of the number of reviews with need of occlusal adjustment (Tab. 2) in the QM group showed that most patients needed 1 adjustment or such procedure was not necessary. In the control group 2 or 3 adjustments were necessary (79%). Presented data show that dentures prepared with the use of an articulator are more physiologic and ensure a balanced occlusion. In the method with the use of an articulator a shorter adaptation period is necessary. A subjective patient estimation of the dentures was also positive for the articulator method. The introduced modification of recording the occlusion ensures an efficient and fast mounting and demounting of the face-bow and its use becomes very advantageous. Semiadjustable articulators should be essential instruments in rehabilitation of edentulous patients. The elaborated procedure is a simple and not time-consuming method. It ensures positive treatment results of edentulous patients assuring all basic aims of masticatory organ rehabilitation. PMID- 9857543 TI - [An attempt to assess the effectiveness of a program for planned dental care during the developmental period in light of clinical examination and questioning of dentistry students from the Pomeranian Medical Academy]. AB - The aim of the study was to assess the effectiveness of planned dental care in educational institutions. The investigation was carried out among 339 3rd, 4th and 5th year students of the Pomeranian Medical University in Szczecin, aged 22 26, who attended nurseries and schools in the years 1969-1989 in different communities (large towns, medium-sized towns, small towns, villages). The first part of the study consisted of questionary answers concerning dental care in general in nurseries, primary and high schools. On the basis of these answers two groups were distinguished: a group of students who had been subjected to planned dental care (positive surveys) and a group of students who had not been subjected to planned care (negative surveys) (Tab. 1). At the second stage the clinical examination of the surveyed students was carried out and the obtained data were registered on 1986 WHO record cards. The examination was performed among students who were subjected to dental care in primary and high school together with those students who were not subjected to this care. All examinations were done at a dental surgery with the use of a mirror, probe and a calibrated WHO-621 probe. Results of the surveys showed that systematically performed examinations of the oral cavity and systematic dental treatment were carried out for over 60% of surveyed students in primary schools and about 40% of surveyed students in high schools. In primary schools the organization of health education was more effective, its activity was confirmed by about 80% of respondents, while in high schools by only 40%. In response to question concerning topical fluoride preventive measures 40% of positive answers were obtained from the surveyed students concerning primary schools and 10% concerning high schools. Survey questions in the part referring to nurseries were often answered "I don't remember" therefore it is impossible to interpret the results. Fluoride tablets were taken by about 40% of surveyed students. Over 65% of respondents with diagnosed malocclusion were treated orthodontically. Among students with dental calculus over a half had it removed earlier. Much better results of clinical examinations were obtained in the group subjected to planned dental care at developmental age. In this group the average DMFT number was 11.31, the average number of teeth with untreated caries (D) was 0.82, average number of removed teeth because of caries (M) was 0.55, whereas in the negative group (not subjected to dental care) the average DMFT number was 14.61, average D number was 1.41, average M number was 2.54. The average number of filled teeth (F) showed no significant difference in both groups and was about 10 (Tab. 2). Malocclusion was diagnosed in about 30% of surveyed students in the group subjected to treatment and in over 45% of surveyed students not subjected to dental care in school (Tab. 2). In the group of positive surveys the proportion of respondents with healthy periodontium was over 11% whereas in the group of negative surveys it was about three times lower (Tab. 4). The results of the study show that planned program of dental care is most effectively carried out in primary schools. The results have confirmed that systematic dental care at development age has a positive effect on the stomatognathic system at mature age. PMID- 9857544 TI - [Evaluation of oral health in the adult population from ages 35-44 in Poland]. AB - The aim of the paper has been to provide objective data comparable to world literature, on the status of masticatory organ in the population of adults aged 35-44 years in Poland. As many as 1380 subjects were studied in 9 voivodeships with due consideration to three inhabited environments: big town, small town and village. The studies were performed in artificial lighting, using a probe and dental mirror, on the basis of methodology found in the WHO instruction of 1986 (Fig. 1.). The status of periodontium and the therapeutic requirements in that respect were assessed by resorting to CPITN index and WHO 621 periodontologic probe. The data concerning the status of dentition reveal a high frequency of caries, claiming 99.8%, as well as high intensity of caries expressed by mean number of DMF equal to 18.6. Despite the fact that the provided index values are comparable to values reached by the majority of developed countries, in case of Polish population aged 35-44 years there is, unlike those countries, very unfavourable distribution for components of D, M and F. The highest value (10.5) is requested by the mean number of missing teeth (M), by mean number of carious teeth (D) with 4.1, while, unfortunately, the lowest being the mean number of filled teeth (F), which is hardly 4.0. So high means of numbers for missing teeth and carious ones indicate that despite the low percentage of toothless subjects in the studied population (1.0%), it will be impossible to reach the 4th health target launched by WHO in the year 2000, it means a 50% reduction in the number of toothless subjects aged 35-44 years. It is evident from data concerning the status of dentition that in the oral cavity of each subject studied there were on the average 21 teeth whereof only 37.2% were made up of healthy teeth. The presence of healthy teeth was recorded more frequently in mandible than in maxilla and most frequently these were incisors and lower canines. Missing teeth (regardless of cause) were disclosed more frequently in maxilla than in mandible. The analysis of the missing teeth according to individual group of teeth reveals their very unfavourable distribution primarily in the lateral segments, it means in zone of support of the masticatory organ. That results in its far reaching consequences with regard to parodontial diseases and temporo-mandibular joints. The established results of studies covering parodontium disclose that it is unrealistic to achieve the 2nd periodontological target of health fixed by WHO for the year 2000, assuming that 75.0% of population aged 35-44 years should possess at least 3 healthy sextants, while deep periodontal pockets (6 mm) were detected in 15.0% of the examined subjects. The poor status of parodontal health is accompanied by colossal need of treatment in this respect. Almost 100% of subjects required instructions with regard to brushing as well as professional procedure in removing dental deposits, whereas 15.0% of the studied demanded specialistic periodontal treatment. The health status of parodontium and the therapeutic demands in this respect for Polish population are also unfavourable against the background of the population residing in the developed countries. The considerable dental losses revealed in the investigation exerted an influence on the prosthetic state of requirements closely related to the above finding. The analysis of data points to high percentages of persons not using removable dentures, particularly the lower ones. It was simultaneously noted that women were more frequent users of movable dentures both upper and lower. No therapeutic requirements involving the upper removable denture were evidenced in a higher percentage of the studied subjects than in the case of lower movable dentures. More persons asked for partial upper denture than lower. As concerns complete denture the situation is found to be reversed. It appears from the data concerning the temporo-mandible joints that in 70.6% of PMID- 9857545 TI - [The influence of acute hypercapnia on the permeability of the blood-brain barrier for gentamycin under conditions of general anesthesia in rabbits]. AB - The aim of the work was to demonstrate whether acute hypercapnia (paCO2 > 65 mm Hg) influenced the permeability of blood-brain barrier (BBB). Twelve Chinchilla rabbits which underwent general anaesthesia were randomly divided into 2 groups. The animals were sedated with intravenous administration of pentobarbital, then were subjected to endotracheal intubation and connected to volume-controlled respirator (Zimmermann pump). Artificial ventilation using air/oxygen mixture was applied. Auricular artery, inferior caval vein and aorta were catheterized with a catheter being also placed in the lateral ventricle of the brain. General anaesthesia was supported with continuous intravenous administration of pentobarbital. To maintain normal paCO2 values, the investigation was performed under normal ventilation in control group (5 rabbits). Controlled hypoventilation was applied to achieve an increase of paCO2 in the shortest possible time in the investigated group (7 rabbits). Heart rate (HR), systolic (SAP), diastolic (DAP) and mean (MAP) arterial blood pressure, intracranial pressure (ICP) and cerebral perfusion pressure (CPP) were continuously recorded. Gentamycin was applied as the marker of function of BBB, because it couldn't penetrate into the cerebrospinal fluid after intravenous administration under physiological conditions. BBB function in normal and significantly increased paCO2 was evaluated using gentamycin permeability indexes (QG), defined as gentamycin concentration ratio in the cerebrospinal fluid to serum gentamycin concentration in the same moment of trial. Comparative analysis of the QG index for both groups according to values achieved before the trial and after 1 and 3 hours of experiment indicates the degree of BBB damage. Non-parametric differences significance test according to Kolmogorow-Smirnow was applied for statistical verification of the results. Significance level for the trial was alpha = 0.05. RESULTS: None of the monitored parameters has changed in the control group. In the group of hypoventilated animals paCO2 values significantly statistically increased and pH values decreased after 1, 2 and 3 hours of the experiment (Fig. 1). In the investigated group increasing paCO2 values were followed by increase of ICP values, which became statistically significant during the third hour of hypercapnia. No statistically significant changes of the CPP values between animals from the control and investigated groups have been noticed. While comparing mean values of the gentamycin permeability index for the investigated group a minimal, statistically insignificant increase of the indexes has been stated during the first hour of hypercapnia. Continuous severe hypercapnia during next two hours caused increase of mean QG value of over 130% (Tab. 1, Fig. 2). That increase remained statistically insignificant as compared to the initial value and also to the estimated one after the first hour of hypercapnia. While comparing QG indexes between control and investigated groups in the given moments of trial it has been stated that mean QG value is statistically significantly higher in animals hypoventilated for 3 hours than in animals which experienced normal ventilation during the whole experiment. CONCLUSIONS: 1. Acute, severe hypercapnia disturbs the function of the blood-brain barrier for gentamycin. 2. For paCO2 values higher than 65 mm Hg no distinct relation between continuous increase of hypercapnia and functional status of blood-brain barrier has been found. 3. Evaluation of the blood-brain barrier function is a new and effective method for gentamycin application. PMID- 9857546 TI - [The effect of deficiency of selected bioelements on hyperactivity in children with certain specified mental disorders]. AB - The aim of my work was the answer to the following questions: how often does the deficiency of magnesium, copper, zinc, calcium, iron occur among hyperactive children in comparison with healthy children, deficiency of which of the considered bioelements is the most frequent, what is the effect of supplementation of deficit element on hyperactivity and does it depend on other certain disorders that coexist with hyperactivity? In a process of establishing the subject diagnosis I have followed the DSM IV criteria recognizing ADHD among examined ones. I have determined the deficiency of magnesium, copper, zinc, calcium, iron in the group of 116 children with diagnosed ADHD. Consequently, as a result, I have found out that shortage of above-mentioned bioelements occurs more often among hyperactive children than among those being healthy, and deficiency of magnesium is the most frequent in this respect. Further, I have divided the group of 110 children with magnesium deficiency into two groups according to the other mental disorders that coexist with ADHD: 1) the group where hyperactivity coexists with disorders typical for developmental age such as enuresis, tics, separation anxiety, stuttering, selective mutism (63 children); 2) the group where hyperactivity coexists with disruptive behaviour disorders: conduct disorder and oppositional defiant disorder (47 children). The content of magnesium, copper, zinc, calcium, iron has been determined respectively in blood (serum and red cells) and in hair by atomic absorption spectroscopy method in both groups accordingly. At the same time, the hyperactivity tests were carried out using Conner's Rating Scales for Parents and Teachers, Wender's Scale as well as Quotient of Development to Freedom from Distractibility. During the statistical analysis the inparametric tests have been used taking as a significance level p < 0.05. On the ground of obtained findings I have not stated any significant differences in bioelements content among hyperactive children in relation to other coexisting disorders, except for zinc. The zinc content in hair was higher among children with ADHD and disruptive behaviour disorder. The assessment of hyperactivity indicated the remarkably higher coefficient among children with coexisting behaviour disorders as compared to hyperactive children among whom, additionally, disorders typical for developmental age have occurred. The analysis of influence exerted by magnesium supplementation on hyperactivity has been carried out in the group of total 75 children with ADHD jointly with magnesium deficiency. The group of 50 children actually tested, apart from standard treatment have received the specified doses of magnesium preparations for 6 months on regular basis. The group of 25 children was left with standard treatment without additional magnesium. In both above-mentioned groups the content of bioelements and respectively ADHD level have been determined just before and after the test. The obtained results have clearly disclosed significant increase of magnesium, zinc, calcium content (Tab. 1) and respectively essential decrease of hyperactivity in the group of children treated with magnesium. At the same time, however, among the children given standard treatment without magnesium, hyperactivity has intensified (Tab. 3, 4). The findings herein presented indicate that it is necessary to take into consideration a possible bioelements deficiency among children with ADHD. Consequently, the accomplished study proves that there is a need of magnesium supplementation in ADHD children irrespectively of other mental disorders. The supplementation of that kind of magnesium supplementation together with standard traditional mode of treatment gives us the opportunity to extend the methods of therapy of ADHD children who are the "children of the risk" in connection with their educational, emotional and social problems. PMID- 9857547 TI - [Tuberculosis of the spine: diagnosis and treatment]. AB - The diagnosis and treatment for tuberculosis of the spine, based on own material from years 1992-97 (CMKP Orthopedic Department, Otwock) is presented. One hundred twenty nine patients were treated in these years, 85 of them surgically. The disease appeared usually in 4th, 5th or 6th decade of life, two adjacent vertebral bodies within lower thoracic spine were involved. Contemporary surgical methods as well as approved antibacterial treatment protocols are presented. Twenty-five per cent patients displayed symptoms of neurological impairment, namely paresis or plegia of the lower extremities. Good results of treatment in 85% of cases and only 3% poor results prove efficiency of the management suggested. PMID- 9857548 TI - [Idiopathic scoliosis: epidemiology and etiology]. AB - Authors present current knowledge about incidence and etiologic factors in idiopathic scoliosis (IS). Most data about incidence are based on screening examination in school children. Amount of patient with IS is related to the method of evaluation and experience of examiner. There is 1.9% to 3% cases in whole population if scoliosis is assumed as much as 10 degrees of Cobb angle (SRS). Etiology of IS is still in doubt despite of many investigations. There are many abnormalities in tissues but most of them probably secondary to this process. Etiology is multifactorial with dominance of inheritance. There are many genes responsible for occurrence of IS but genetic trait is still unknown. PMID- 9857549 TI - [Nonoperative treatment of acute, grade III acromioclavicular dislocation in judo competing athletes]. AB - Complete acromioclavicular separation is frequent injury during practice of judo. Fourteen athletes aged 17-33 (mean 23) were treated non-operatively. All patients received cryotherapy and Velpeau bandage for two weeks, intensive rehabilitation of the shoulder followed. The patients were reviewed after mean 4.5 years (range 3-7 years). A 100-point scale designed specifically for this study, inclusive of pain, function, strength, range of motion and return to preinjury level of sports activity was used for clinical assessment. Follow-up radiographs were compared to those taken immediately after injury but no correlation with clinical results has been found. Eight results were rated excellent (90-100 points), 5 results good (80-89 points) and one result was fair (70-79 points). Short period of immobilization and early rehabilitation seems to be efficient management in highly active individuals. PMID- 9857550 TI - [Remarks on Gamma nail use in the treatment of trochanteric fracture of the femur]. AB - Results of Gamma nail use in treatment for trochanteric fracture of the femur in 25 patients (15 females, 10 males) aged 45-91 years are presented. There were 12 type I, 9 type II, 3 type III and 1 type IV fracture according to Boyd-Griffin classification. Fracture healed 10-12 weeks after surgery in 24 cases; one patient died because of pulmonary embolism. PMID- 9857551 TI - [Clinical and radiological assessment of proximally coated stems in growth in total hip replacement]. AB - Clinical and radiological results of total hip replacement with proximally coated stems are discussed. Material included 90 hips (50 ABG stems, 40 Level III stems) in 83 patients (59 females, 24 males), mean age 48 years (range 17-22) and mean follow-up of 27 months (range 1-5 years). No stems were revised, the average Harris Hip Score at last follow-up ranged from 89 to 91 points. Stress transfer around the proximal part of the stem resulted in endosteal osteogenesis in the 2nd and 6th Gruen zones and changed trabecular orientation. There was no evidence of bone resorption due to stress shielding. Peri- and endosteal bone reaction in the 3rd and 5th Gruen zones need longer follow-up. PMID- 9857552 TI - [Mecring threaded cup in total arthroplasty of the hip with acetabular protrusion]. AB - Results of total arthroplasty of the hip with acetabular protrusion with the use of Mecring cup in 21 patients (25 hips) aged 25-75 years (mean 62) are reported. Morsalised impacted autografts were used to fill acetabular defect in all cases. There were 14 rheumatoid arthritis patients, 4 cases of osteoarthritis and 3 cases of Otto-Chrobak disease. Mean follow-up was 2.5 years (range 6 months-5 years). All autografts were incorporated after 6-9 months. Only one loosening and migration of the cup occurred. One patient died because of pulmonary embolism. Harris Hip Score increased from 37 points (22-49) preoperatively to 81 points (65 89) at the latest follow-up. Incorporation of autografts reinforced acetabular bottom and prevented recurrence of protrusion. Bone remodeling indicates restoring normal biomechanics of the hip. PMID- 9857553 TI - [Distal femur reconstruction with vascularized fibular graft: indications and surgical technique]. AB - Indications and surgical technique of distal femur reconstruction with vascularized fibular graft and fibular vessels pedicle are presented. The method is used in femoral metaphyseal defects combined with knee stiffness as safer procedure than free vascularized fibular graft. A case of 23 years old male with vast defect of distal femur after fracture complicated with osteomyelitis is presented. Good result of treatment has been achieved 18 months after surgery despite fracture of previously incorporated fibular graft. PMID- 9857554 TI - [Is posttraumatic osteomyelitis of iatrogenic nature?]. AB - A series of 112 patients with posttraumatic osteomyelitis has been reviewed. Analysis of the management revealed, that malpractice contributed to the onset of osteomyelitis in many cases. Since politrauma patients with extensive bone and soft tissue injury are prone to infection errors in management of these patients lead to serious complications. Neglecting of primary immobilization of the open fracture in politrauma patients, incorrect methods of fracture fixation or technical errors at surgery, erroneous soft tissue injury management (complete closure of the wound, neglecting compartment syndrome) and disregarding local antibiotics are among most frequently committed errors. The authors conclude, that in majority of cases posttraumatic osteomyelitis is of iatrogenic nature. PMID- 9857555 TI - [Osteoid osteoma of the lower extremity]. AB - Forty patients (27 males and 13 females) aged 18 months-41 years with preliminary diagnosis of osteoid osteoma localized within lower limb were reviewed. All underwent surgery. Histopathology confirmed previous diagnosis in 29 patients but in 11 cases (30%) histology either gave different diagnosis (despite clinical and radiological changes typical for osteoid osteoma) or was inconclusive. Three types of radiographic picture were distinguished: within cancellous bone the change was either osteolytic or oval, sclerotic; within cortical bone it was spindle-shaped, sclerotic. In 5 cases radiological symptoms were present prior to the pain. In 3 cases observed several years before surgery natural growth of osteoid osteoma could be traced. The key role of prostaglandin synthesized by osteoid osteoma in its pathology was emphasized. PMID- 9857556 TI - [Anthropometric parameters in assessment of patients with Marfan syndrome or with Marfan phenotype]. AB - A series of 37 patients aged 4-64 years has been evaluated with criteria of Lee and Ramirez. Diagnosis of Marfan syndrome has been established in 13 cases, in 24 patients the Marfan phenotype has been found. In both groups body height, upper extremities length, the length of upper and lower body segment, length of the foot and hand have been recorded. Metacarpal index has been calculated. Antropometric measurements did not reveal significant differences in body parts proportions between these two groups. PMID- 9857557 TI - [Anterior knee pain syndrome: a historical review]. AB - Anterior knee pain is a frequent musculoskeletal complaint affecting adolescent population with incidence of 36% in 14 years old schoolchildren. Until late sixties anterior knee pain used to be associated with chondromalacia. This review discusses etiology and treatment of anterior knee pain. Various etiologic theories exist ranging from trauma and patellar maltracking to retinacular nerves injury. None of these theories has been generally accepted and etiology of patellofemoral pain remains unclear. PMID- 9857558 TI - [Treatment of benign chondroblastoma]. AB - Six children have been operated on due to benign chondroblastoma at the mean age at first operation of 14.2 years (range 11-17 years). Mean follow-up was 4 years (2-7 years). Mean delay of the diagnosis was 12 months (6-24 months). One-stage surgery was successful in child with proximal fibula involvement, child with distal femoral epiphysis involvement (knee arthrodesis followed), two children with proximal tibia involvement. In 1 case of distal femoral epiphyseal involvement two revisions were necessary, 1 similar case is in the course of treatment. At the follow-up three children presented with the knee pain and restricted range of motion. PMID- 9857559 TI - [A case of infected tibial pseudoarthrosis treated by Ilizarov method complicated by supracondylar traumatic fracture of the femur]. AB - A case of 45 years old male with infected pseudoarthrosis of the tibia, 4 cm shortening of the limb involved, complicated with ipsilateral distal femur fracture is reported. Ilizarov apparatus was used to address whole pathology. At 8 months follow-up good clinical and radiological result is achieved--the fracture and pseudoarthrosis united, the leg is lengthened by 4 cm. PMID- 9857560 TI - [Osteopetrosis: a case report and review of literature]. AB - A case of type II autosomal dominant osteopetrosis is presented. The boy was admitted to the hospital at the age of 7 years and 6 months due to bilateral leg pain. Apart from radiography no pathology has been found. PMID- 9857561 TI - Cyclic changes in genital organs and vaginal cytology in cynomolgus monkeys (Macaca fascicularis). AB - Vaginal cytology was evaluated weekly over 12 months in 20 adult female Cynomolgus monkeys (Macaca fascicularis). After sacrifice of the animals the histology of the ovaries, uterus and vagina were studied in different phases of the menstrual cycle. The cytological examination of the vaginal smears showed that the superficial cells increased in number towards the middle of the cycle and the number of intermediate cells declined gradually. Parabasal cells were observed mainly at the beginning of the cycle; they disappeared towards the middle of the menstrual cycle. During the early follicular phase, the cells were moderately separated from each other, and during the second half of the proliferative or follicular phase, the superficial cells appeared clumped together. Leucocytes were usually absent except for at the beginning of the cycle and in the last few days of the late secretory or luteal phase. The maturation index of the vaginal smears can be considered as a tool for distinguishing the different phases of the menstrual cycle. The microscopic examination of the genital organs showed that during the proliferative or follicular phase of the cycle, which corresponds to the development of the ovarian follicles, the uterus showed growth of endometrial glands, stroma and endothelial cell proliferation with capillary sprouts. Shortly after ovulation and parallel to the formation of the corpora lutea, the endometrium enters the secretory or luteal phase, which is characterized by coiling of endometrial glands, glandular secretion and the differentiation of the spiral artery. The most striking changes in the vagina, is the marked basal cell proliferation and thickening of the stratum granulosum during the follicular phase of the menstrual cycle. The histological changes observed in the vagina demonstrated a good correlation with the observation on cytological examination of the smears. The present study demonstrated that the process of angiogenesis in the uterus during the different phases of the menstrual cycle is a multiple phenomenon involving proliferation, maturation and differentiation. PMID- 9857562 TI - [Effect of the evaluation parameter on sensitivity of resonance thrombography of thrombocytopenia in dogs]. AB - Based on 109 blood samples taken from 36 dogs suffering from thrombocytopenia resonance thrombography with the resonance thrombograph RTG 801 (von Hoerner und Sulger Electronic GmbH, Schwetzingen; manufacturer: Fresenius AG, Bad Homburg) was distinctly more sensitive and more closely correlated to the platelet count using an optimized parameter of the resonance thrombogramm (RTG) in comparison to usual parameters. Nevertheless, clinical requirements regarding samples with platelet counts > 25,000/microliter were not fulfilled. Out of 13 samples with reduced platelet count and simultanous extended capillary bleeding time, depending on the used parameter a maximum of 9 samples could be detected as pathological by the RTG. The normal RTG in part of the cases with clearly altered primary haemostasis contrasts to the exclusive use of RTG in the screening of thrombocytopenia in dogs. PMID- 9857563 TI - [Possibilities and limitations for use of the polymerase chain reaction (PCR) in the diagnosis of bovine leukemia virus (BLV) infection in cattle]. AB - Enzootic bovine leukosis is caused by the bovine leukemia virus (BLV) and has a world wide distribution in cattle. Due to the program for eradication of BLV infections in Germany the BLV incidence in cattle declined and only few new cases seem to occur per year. On the other hand, BLV-infected cattle with low, transient or without BLV-antibody titers are difficult to identify as BLV infected. These animals may be sources for new infections. It was the aim of this study to compare the suitability of agargel-immunodiffusion (AGID), enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) for diagnosis of BLV-infected cattle. We investigated a herd with 10 cows, where after a long period when the herd was negative suddenly a positive serological reaction appeared. In addition 64 animals from 6 federal states of different herds with doubtful serological reactions found in previous tests were included. In the herd with 10 cows we were able to detect BLV-infection in one animal 8 weeks earlier with PCR than with ELISA. Investigation of 56 adult cattle and 3 calves from different herds with both PCR and ELISA showed that 51 animals were positive in ELISA and 55 in PCR. Seven animal were positive in PCR and negative in ELISA. Three calves yielded negative results in PCR and positive results in ELISA. One cow which was positive in previous serological tests was negative in ELISA, AGID and PCR. Restriction fragment length polymorphism analysis demonstrated that the majority of the cattle was infected with the same BLV provirus variant. The four PCR variants used in this study yielded a similar sensitivity for BLV provirus detection. In conclusion, compared to the serological tests, PCR detects BLV infection earlier in naturally infected cattle. The method is also a useful tool to exclude or confirm BLV-infection in cattle with doubtful serological results. PCR may be used to complement the serological tests in the diagnosis of BLV infection. PMID- 9857564 TI - [Clinical findings and differential diagnosis in ketosis and hypocalcemia in sheep]. AB - Clinical and laboratory examinations were carried out in a prospective study from 1978 to 1998 on 454 spontaneously diseased sheep of different breeds aged 2-8 years. Three groups of sheep were defined by plasma concentrations of 3 hydroxybutyrate (3-OHB) and calcium. 214 ewes suffered from ketosis (pregnancy toxaemia, 3-OHB > 1.6 mmol/l), 195 ewes and 5 rams suffered from hypocalcaemia (Ca < 2.0 mmol/l), and 40 ewes were sick with booth diseases simultaneously. Clinical findings differ only slightly in all three groups. In ketosis the time until the onset of clinical signs and the duration of the disease is somewhat longer, while locomotion disturbances are less severe than in hypocalcaemia and in the combination of both. In individual cases a differential diagnosis is not possible on the basis of clinical findings, and also acetonuria is not a reliable criterion. Therefore, a therapy is suggested to treat all three possible forms of disease: daily oral application of Na-propionate, Ca-lactate and K-chloride, and additional subcutanous injection of Ca-borogluconate, vitamin D3, alpha Tocopherol and selenium at the beginning of the disease. Pathogenesis of both diseases are discussed, recommendations for feeding management are given. PMID- 9857565 TI - [Endoparasitic infections in sheep from the Swabian Alb]. AB - The endoparasite fauna of 59 slaughtered sheep (30 lambs, 29 ewes) from the Swabian Alb, Germany, was examined. One species of trematodes, 3 species of cestodes, 29 species of nematodes (23 species of gastro-intestinal and 6 species of lung nematodes), 1 species of arthropodes and 1 species of protozoa were recorded. All animals were infected with Dicrocoelium dentriticum as well as gastro-intestinal and lung nematodes, 45.8% with Moniezia spp., 15.3% with Cysticercus tenuicollis, 55.9% with Oestrus ovis and 11.9% with Sarcocystis gigantea. The most important gastro-intestinal nematodes were Ostertagia circumcincta and Cooperia curticei, which were recorded in all sheep, Ostertagia trifurcata and Chabertia ovine (98.3% each), Oesophagostumum venulosum (96.6%), Nematodirus filicollis (81.4% each), Ostertagia pinnata (78.0%), Trichuris ovis and Trichostrongylus colubriformis (76.3% each). The ewes harboured more abomasal and small intestinal nematodes (1819 and 3702) than the lambs (695 and 1730), which haboured more large intestinal nematodes (177) than those (56). The most often recorded lungworms were Cystocaulus ocreatus (74.6%) and Muellerius capillaris (72.9%), followed by Neostrongylus linearis (57.6%), Dictyocaulus filaria (50.8%), Protostrongylus brevispiculum (37.3%) and Protostrongylus rufescens (28.8%). The ewes carried higher lungworm burdens than the lambs. PMID- 9857566 TI - Studies on laticiferous plants: toxic effects in goats of Calotropis procera latex given by different routes of administration. AB - The effects on goats of Calotropis procera latex given by different routes of administration were investigated. The administration of latex at 1 ml/Kg body weight via the oral route or at 0.005 ml/Kg body weight/day via the intravenous or intraperitoneal route caused death of the goats between 20 minutes and 4 days. When the small dose of latex (0.005 ml/Kg body weight/day) was given by the oral route or intramuscular route no death among the goats occurred. Nervous signs, frequent urination, frothing at the mouth, dyspnoea and diarrhoea were the main features in goats given latex by the oral, intravenous or intraperitoneal route. Lameness was observed in goats given latex via the intramuscular route. Lesions were widespread congestion and haemorrhage, pulmonary cyanosis, enterohepatonephropathy, peritonitis (in goats receiving latex via i.p. route) and haemorrhagic myositis at the site of latex injection. These changes were accompanied by increases in the activities of serum GDH, LDH, ALP, GGT and AST and in the concentrations of cholesterol, urea and creatinine and decreases in the level of total protein. PMID- 9857567 TI - The hypoglossal nuclear complex in reindeer. AB - The hypoglossal complex has an ovoid shape and is largest at the level of obex. Caudally to the obex is 2/3 of the complex. The complex is divided into 4 groups. Sublingual nucleus of Roller presents a narrow strip of cells located ventrally to the middle part of hypoglossal nucleus. Laterally to obex is the paramedian nucleus of Jacobson in the form of short, narrow band. Posteriorly to Jacobson's nucleus is the intercalated nucleus of Staderini. Constinuation of the hypoglossal nucleus is the nucleus prepositus of Marburg. PMID- 9857568 TI - Morphometric study of the human embryonic retina. AB - In 33 embryos the retinal thickness and neuronal density was investigated. Diameter of the eye in relation to embryonic age increases linearly to stage 20 and after this period is more pronounced. Thickness of the external nuclear layer increases slowly in the investigated period. The internal nuclear layer appears at stage 17 and in increases in thickness fourfold to the end of embryonic period. PMID- 9857569 TI - Morphometric study of the muscles spindles of the eye muscle in human fetuses between 24-29 weeks. AB - The studies were carried out on human fetuses with the crow-rump length between 230 to 270 mm (24-29 weeks). The length of recti muscles varies from 12-15 mm in 24th week to 25-28 mm in 29th week. Number of muscle spindles varies from 29-36 in 24th week to 54-59 in 29th week. The length of the muscle spindles is from 30 300 microns in 24th week to 40-450 microns in 29th week. The diameter of the muscle spindles varies from 11-36 microns in 24th week to 15-70 microns in 29th week. PMID- 9857570 TI - The orifice of superior thyroid vein to the internal jugular vein in human fetuses. AB - In this study an attempt was made to define the ways of the orifice of the superior thyroid gland to the internal jugular vein in human fetuses, basing on corrosion casts of blood vessels. It was found that most frequently, i.e. in 44% of cases, the superior thyroid gland formed bilaterally multi-level anastomoses with the facial vein and lingual vein before the orifice to the internal jugular vein. In addition, anastomoses of the three mentioned veins into a common venous trunk and non-anastomotic course of the superior thyroid vein were found. PMID- 9857571 TI - Variations of the anterior communicating artery. AB - Study was performed in 30 human cadavers during autopsy. The anterior communicating artery was present in all investigated cases. In 30% of instances the artery was of diameter less than 1 mm (hypoplastic). PMID- 9857572 TI - Suggested nomenclature for terminal orders of bronchial ramifications and segmental arteries in the upper lobe of the left human lung. AB - Official nomenclature of bronchi and of pulmonary arteries includes segmental and some subsegmental branches. However, progress in clinical practice requires that nomenclature of subsequent order ramifications should be introduced. Nomenclature of the branches suggested by other authors is presented. Analysis of pulmonary artery branching within the upper lobe of the left lung, made on 150 corrosive preparations, and analysis of radiograms has prompted the author to suggest his own notation for the fifth and sixth order branches. PMID- 9857573 TI - Variations of the superficial palmar arch. AB - The superficial palmar arteries were carefully dissected, using gross techniques, in 80 conserved cadaver hands. 78 of the specimens (97.5%) were shown to have the superficial palmar arch 64 (80%) of which were formed by anastomoses between ulnar and radial arteries. In 14 of the specimens (17.5%) the arch was formed by the ulnar artery alone. There were no arches in 2 of the specimens (2.5%) which were supplied by the anterior median artery. PMID- 9857574 TI - Estimation of health state and physical efficiency of children from rural area of Lubin-Glogow copper-basin and lead levels in blood serum. Preliminary results. AB - The aim of the work was to estimate the health state and physical efficiency of children from rural area of Lubin-Glogow Copper-Basin in correlation with lead levels in blood. Lead concentration in blood showed increase tendency together with age of the children. In both sexes together with rise in lead concentration in blood enlargement of palatine tonsils was observed. In children with a higher lead concentration in blood lower values of physical fitness parameters are recorded. PMID- 9857575 TI - Change of some values of fetal metric features during storage and preservation. AB - On the material of 127 human fetuses of both sexes aged 84-112 days of fetal life as well as on 127 fetuses aged 228-256 days of human life, body length (BL), body mass (BM), head circumference (HC) and thorax circumference (ThC) were measured before their preservation. Value changes of these features were assessed as they are exposed to the following factors: placing in water for 6, 12 and 24 minutes; storing in refrigerator and control measurements were done after 12, 24 and 36 hours; storing in 10% solution of formalin for 1 month, 3 month and after 1 year. PMID- 9857576 TI - Mastoid emissary in Indian skulls. AB - Two hundred and eleven Indian skulls originating from medieval times, South of Peru, were studied in respect to variability of the mastoid emissary. Occurrence and localisation of the external orifice of the mastoid emissary were studied in three distinct and well-separated populations, living in the three different places: Villa el Salvador, Tablada de Lurin and Paracas. Most of the skulls presented considerable degree of artificial deformation. External orifice of the mastoid emissary was multiple in most of the skulls, especially in Villa el Salvador and Paracas populations. Mastoid emissaries in Indian were situated generally higher than in European, especially regarding the Paracas population. Significant sex differences were present in this material, regarded as a whole (summarized results for the three populations), what is in agreement with previous observations on European skulls. It indicates that even in cases of relatively deep deformation, some characteristic features of sex dimorphism of the human skull are noticeable. Additionally, the three studied populations, which were well separated from each other and lived alone, differed significantly regarding localization of the mastoid emissaries. PMID- 9857577 TI - Jens Peder Hart Hansen. 3 November 1936-23 September 1998. PMID- 9857578 TI - Substance abuse-high priority for public health. PMID- 9857579 TI - Variability of substance abuse. Global variability of substance abuse: is latitude a unique etiological factor? AB - Worldwide substance abuse consequences are a major problem challenging health planners and providers. To mediate these problems effectively, further information on the variability of substance abuse prevalence and associated causes is needed. There is some evidence suggesting that latitude may present unique etiology for substance abuse because of northerly conditions such as extreme light and dark cycles and longer periods of cold harsh environment. This hypothesis is investigated by reviewing the known literature and applying methods for evaluating latitude as a geophysical grouping characteristic on archival substance abuse data. Conclusions are based on previous findings and examples of alcohol-attributable mortality for populations from six northern areas and the United States. PMID- 9857580 TI - Persistent organic pollutants in maternal blood plasma and breast milk from Russian arctic populations. AB - Under the auspices of Arctic Monitoring and Assessment Programme (AMAP), a Russian-Norwegian co-operation project was established to assess the exposure of delivering women to persistent organic pollutants (POPs) in Arctic areas of Russia. In the period 1993-95 blood and breast milk samples were collected from 94 delivering women in Yamal and Tajmyr Autonomous Regions of Siberia. Concentrations of chlorinated pesticides and polychlorinated biphenyls (PCBs) were determined by high resolution gas chromatography with electron capture detection. The POP levels in maternal plasma among the non-indigenous women were higher than the native population, especially in total PCB, HCHs (hexachlorocyclohexanes) and the DDT-group. The dietary questionnaires showed that the non-indigenous populations consumed considerably less local food items like reindeer meat and fresh water fish. There was no correlation between local food consumption and elevated levels of pollutants. Even if the indigenous groups had lower concentrations of the most important pollutants than the non-indigenous population, they were still higher than the levels measured in the Scandinavian countries of the AMAP-study and up to levels of medical concern. The most important sources of organic pollutants for the Russian Arctic populations of Yamal and Tajmyr seems to be imported food from other areas of Russia and local use of pesticides. It must be a high priority concern to further elucidate these trends and initiate prophylactic measures for the exposed population groups. PMID- 9857581 TI - Building-related health problems: reflections on different symptom prevalence among pupils and teachers. AB - The occurrence of non-specific symptoms among populations in modern buildings is common in the northern parts of the world, and is often called the Sick Building Syndrome. Many factors have been shown to be associated with the prevalence of such symptoms. Based on a case study of a primary school in subarctic Sweden, the complicated nature of building-related, non-specific symptoms is reviewed. Preventive and corrective actions in cases of sick buildings often fail. It is suggested that failures of such actions might depend on their predominant origin in biomedical models. The study therefore proposes that the combined and simultaneous use of biomedical and psychosocial models should be tried in the management of building-related health problems. PMID- 9857582 TI - Place of the Hokkaido Ainu (northern Japan) among circumpolar and other peoples of the world. A comparison of the frequency variations of discrete cranial traits. AB - Fifteen discrete cranial variations were studied in Ainu and a number of populations from around the world. The major findings, extracted by B-squared distance analysis and Fisher's exact probability test, are as follows: A) The five key traits which distinguish Ainu from the predominant eastern Asians are (1) medial palatine canal, (2) hypoglossal canal bridging, (3) supraorbital foramen, (4) transverse zygomatic suture vestige, and (5) mylohyoid bridging. The frequency of the first trait is comparable to those of Africans, especially Subsaharan Africans, the second and fifth conform to those of the New World peoples, the third is similar to Australians and Subsaharan Africans, and the fourth is the only one aligning the Ainu to Northeast Asians. B) The frequencies of a few wormian bones in Ainu are significantly different from those of many Southeast Asians. The frequencies favour rough clinality from the south to as far north as the arctic area of Northeast Asia in the regions of western Oceania and eastern Asia. The present study confirms that Ainu and Jomon are certainly members of populations originated in Asia but possibly outliers of more predominant Asian populations. PMID- 9857583 TI - The course of childhood bronchiectasis: a case report and considerations of hospital use. AB - This paper discusses the clinical history of a Finnish person from the area north of the Arctic Circle with childhood-onset bronchiectasis. It covers a period of 30 years and is aimed at analysing the significance of the disease for the use of hospital services. Childhood bronchiectasis is a rare disease nowadays and difficult to diagnose, but it still leads to serious disability in some children, requiring regular hospitalisation. The prognosis for these cases with advancing age is not known. PMID- 9857584 TI - The human health programme under AMAP. AMAP Human Health Group. Arctic Monitoring and Assessment Program. AB - The human health programme of the first phase of AMAP was planned at an international meeting held in Nuuk, Greenland, October 1992. As the most vulnerable period to adverse effects of contaminants is during fetal development, it was decided to concentrate on analyses of umbilical cord blood and maternal blood. The programme was designed as a core programme in which 150 sample pairs should be collected in each of the 8 arctic countries and analyzed for persistant organic pollutants (POPs) and heavy metals (mercury, lead and cadmium). As some essential elements such as copper, zinc and selenium interfere with heavy metal toxicity these elements should also be analyzed. Additional analyses such as nickel and arsenic in urine, mercury in hair, and POPs in breast milk could be incorporated regionally according to specific local conditions. Radionucleides were not a major focus in the human programme as this issue was be dealt with by AMAP's radiation group. Implementation of the programme was a problem in most of the countries due to lack of funding. However, an offer from Canada to analyze all contaminants in 50 samples from each country enabled the first comparative circumpolar study of human exposure to contaminants to be completed. The study confirmed that in general the most important source of exposure to both POPs and mercury is food of marine origin and that Greenlanders and Inuit from the Canadian Arctic, due to their traditional lifestyle, are among the most highly exposed populations in the Arctic. This is not a result of local pollution in Greenland and Canada, but is due to long range transport of persistent contaminants through the atmosphere and their biomagnification in the marine food chain. For these reasons the most important recommendation of the first AMAP assessment is that priority should be given to the expeditious completion of negotiations to establish protocols for the control of POPs and heavy metals under the Convention on Long Range Transboundary Air Pollution. PMID- 9857585 TI - In vivo performance of a modified CSTi dental implant coating. AB - Cylindrical dental implants coated with cancellous structured titanium (CSTi) were studied in a dog model. CSTi-2-coated and hydroxyapatite-coated (HA) implants were placed in 8 mongrel dogs. The porosity of the CSTi-2 coating was 9% less than that of the previously studied CSTi-1, resulting in greatly improved mechanical strength and cosmetic appearance. A slightly lower level of bone ingrowth was observed for CSTi-2 than for CSTi-1. However, the in vivo attachment strength of the CSTi-2 coating was comparable both to CSTi-1 and to an HA-coated control after 8 weeks. Measured porosity is technique dependent; digital analysis of in vitro samples yielded higher porosity values than in vivo histology cross sections. PMID- 9857586 TI - Two-stage maxillary sinus reconstruction with endosseous implants: a prospective study. AB - The aim of this prospective study was to evaluate the results of 2-stage maxillary sinus reconstruction using titanium implants placed into iliac corticocancellous bone blocks previously grafted to the floor of sinuses. Fifty consecutive patients received 314 Branemark implants of varying lengths; 202 implants were placed in the grafted bone and 112 were placed in the adjacent anterior maxillary alveolar process, which had received buccal onlay bone grafts. Follow-up time was 9 to 48 months after implant placement, which was accomplished 5 months after bone grafting. Eighty-four percent of the implants were integrated into the grafted sinuses and 75% were integrated into the anterior graft. Six patients (12%) lost implants in strategic positions, leading to secondary implant placement prior to fabrication of fixed prostheses. Thirty-eight patients (76%) received fixed prostheses. Only 5 individuals (10%) attained permanent implant anchored overdentures. One patient lost all implants. The total implant survival rate (80.9%) and the survival rate of the fixed prostheses (100%) compare favorably with other reports. PMID- 9857587 TI - Endosseous implant and autogenous bone graft reconstruction of mandibular discontinuity: a 12-year longitudinal study of 31 patients. AB - Surgical, medical, and prosthodontic records of 61 consecutively treated patients with mandibular discontinuity were reviewed retrospectively. All 61 patients had undergone discontinuity reconstruction with autogenous bone grafts; 31 of 61 had also received endosseous dental implants and a dental osseoprosthesis. Of these 31 implant-reconstructed patients, 23 had free autogenous nonvascularized and 8 had vascularized bone grafts. The surgical-prosthetic protocol consisted primarily of secondary, free autogenous nonvascularized bone graft reconstruction and secondary root-form endosseous implant and fixed prosthesis dental reconstruction. Vascularized bone (8 patients) or soft tissue (4 patients) grafts were utilized selectively for severely compromised patients after extensive oncologic resection, avulsive trauma, or after previous radiation treatment. Endosseous implant survival (95.5% in 31 patients), autogenous bone graft success (98.4% in 61 patients), and dental osseoprosthesis success (100% in 31 patients) were favorable. A high incidence (9.1%) of nonfunctioning (sleeping) implants was recorded for this patient population. The need to remove the titanium mesh tray for various reasons (17.6%) and the need to reconstruct soft tissue in the irradiated patient (12%) were noteworthy. PMID- 9857588 TI - Influence of prosthesis material on stress distribution in bone and implant: a 3 dimensional finite element analysis. AB - A 3-dimensional finite element analysis was conducted to assess stress distribution in bone, implant, and abutment when gold alloy, porcelain, or resin (acrylic or composite) was used for a 3-unit prosthesis. A unit force was applied axially and then buccolingually to the center of the pontic. For gold and porcelain, similar maximum equivalent stress was found in each part of the models. In almost all cases, stress in the model with the resin prostheses was similar to or higher than that in the models with the other 2 prosthesis materials. The highest increase in stress with the resins was found in the implant-abutment unit under axial load. The protective role of resin for the implant-bone interface could not be demonstrated under the conditions of this analysis. PMID- 9857589 TI - Short (6-mm) nonsubmerged dental implants: results of a Multicenter clinical trial of 1 to 7 years. AB - Limited bone height restricts the use of long dental implants, so short implants may be selected in these situations. Recent reports on clinical results with short implants have been negative, however, and have suggested that indications for the use of these implants are limited. To verify these findings, a multicenter study of short ITI implants was carried out. In a 6-year period 253 short implants with a length of 6 mm were placed into 126 patients, who were followed up from 1 to 7 years. Altogether 7 implants were removed; 6 of these were located in the maxilla and 1 in the mandible. The quality of survival was comparable with the clinical results of longer implants from the same implant system. Although the clinical results of these short implants were favorable, it is recommended that they be used in combination with longer implants, especially when used in the less dense bone that is often seen in the maxilla. PMID- 9857590 TI - Osteotome single-stage dental implant placement with and without sinus elevation: a clinical report. AB - Forty-three sites in 16 patients were selected for placement of implants using osteotomes in a single-stage surgical technique. Most sites were treated to enhance bone quality. Sixteen sites in the posterior maxilla had sinus elevation performed in conjunction with implant placement. No soft tissue coverage of the implant cover screw was attempted in either the sinus-elevated or the nonelevated sites. Sinus elevation was significant relative to baseline (mean gain 3.25 mm, P < .01). The implant survival rate was 95.3%. PMID- 9857591 TI - Histologic and histomorphometric analysis of the bone response to machined and sandblasted titanium implants: an experimental study in rabbits. AB - The aim of this study was to make a comparative analysis between the bone response to machined and sandblasted implants. The sandblasting was done with 150 microns aluminum oxide particles. Under scanning electron microscopic examination, the machined implants presented typical machining grooves, while a very rough, highly irregular surface with depressions and indentations was present on the sandblasted implants. Light microscopy showed a different bone growth pattern on machined (implantopetal growth) and sandblasted (implantofugal growth) implants. No negative effects on the rate of bone growth were observed in spite of the presence of aluminum ions. The histomorphometric analysis showed that sandblasted implants presented, from the third week onwards, a significantly higher contact percentage (P < .0001). These values could point to higher osteoconductivity as a result of the higher surface roughness of sandblasted surfaces. PMID- 9857592 TI - Measurement of maxillary sinus volume using computerized tomographic images. AB - This study measured maxillary sinus volume as an aid in determining the volume of graft bone needed before grafting autogenous bone to the maxillary sinus floor. Maxillary sinus volumes were measured from computerized tomographic images of 38 sinuses using a 3-dimensional reconstruction system. When the sinus-lift procedure was simulated, volumes (mean +/- SD) of the inferior portion of the sinuses were 4.02 +/- 1.44 cm3 for 15-mm lifting and 6.19 +/- 1.77 cm3 for 20-mm lifting. In bone grafting of the maxillary sinus floor, taking into consideration individual differences in maxillary sinus volume and resorption of the grafted bone, 5.46 cm3 or more were required for a 15-mm lift and 7.96 cm3 or more were required for a 20-mm lift. PMID- 9857593 TI - Full-arch restoration of the jaw with fixed ceramometal prosthesis. AB - Between 1990 and 1995, 214 implants were placed in 29 maxillae and mandibles of 22 patients following extraction of all residual teeth as a consequence of severe periodontal disease. All patients were discharged wearing immediate dentures. The implants were analyzed with regard to the number per arch, location, length, and diameter. The 5-year cumulative survival rate was 98.5%. The mean number of implants per arch was 7.5 for the maxilla and 7.2 for the mandible. The preferred implant locations were canines, central incisors, lateral incisors, and second premolars in the maxilla; and lateral incisors, first molars, and canines in the mandible. The mean implant length was 14.7 mm in the mandible and 14.5 mm in the maxilla. The mean implant diameter was 3.8 mm in the maxilla and 3.8 mm in the mandible. The results of the present study indicate that immediate implantation for fixed full-arch reconstruction can be considered a viable treatment alternative in patients with severe periodontal disease. PMID- 9857594 TI - Effects of extracellular matrix constituents on the attachment of human oral epithelial cells at the titanium surface. AB - This in vitro study attempts to delineate the role of extracellular matrix (ECM) constituents at the epithelial tissue-implant interface. To know which ECM constituents have a beneficial influence on the behavior of epithelial cells, the attachment, proliferation, morphologic pattern, and differentiation or cytoskeletal organization of human oral epithelial cells on ECM-coated (type IV collagen, fibronectin, type I collagen, laminin, and vitronectin) and noncoated titanium surface have been evaluated and compared. In each experiment comparing commercially pure titanium and oxygen plasma-cleaned titanium, the same ECM constituents were used. In this study, type IV collagen could provide an excellent substratum for epithelial cell attachment on titanium surface, but vitronectin-coated titanium revealed lower effectiveness for attachment of epithelial cells than noncoated titanium. These results suggested that type IV collagen could be used as a means for obtaining good epithelial seal, whereas vitronectin could be used to restrain the attachment of epithelium to dental implants. PMID- 9857595 TI - Hot isostatic pressing-processed hydroxyapatite-coated titanium implants: light microscopic and scanning electron microscopy investigations. AB - Hot isostatic pressing (HIP) was used in a new procedure to produce hydroxyapatite (HA) coatings on a commercially pure titanium (cpTi) substrate for osseous implantation. Eighteen HIP-processed HA-coated implants were placed in the inferior border of the mandibles in 2 Labrador retriever dogs and left submerged for 3 months. As control specimens, 12 sandblasted cpTi implants were placed in the same mandibles and, to compare the bone reaction, 2 additional plasma-sprayed HA-coated implants (Integral) were placed. Tissue reactions at the bony interfaces of the implants were studied in ground sections with the implants in situ, using ordinary, fluorescent, and polarized light microscopy and scanning electron microscopy (SEM). The HIP-processed HA coatings displayed an increased density in light microscopy and SEM as compared to plasma-sprayed coatings. Direct bone-implant contact was found in all 3 types of surfaces. However, the production of new bone was far more abundant for the HA-coated implants than for sandblasted cpTi implants. The presence of bone-forming and bone-resorbing cells indicated active bone remodeling in the interface area at 3 months after implant placement. The present results support the view that epitaxial bone growth may occur from the HA-coated implant surface. It was concluded that the increased density of the present HIP-processed HA material does not reduce the bioactive properties of the coatings. PMID- 9857597 TI - Mandibular implant-retained overdenture: a clinical trial of two anchorage systems. AB - This in-vivo study aimed to investigate the load on the working-side implant and on the edentulous distal mucosa of the nonworking side in a mandibular implant retained overdenture (MIR-OVD) anchored to 2 implants by either a ball- or a clips-and-bar attachment. Three female patients were provided with duplicate dentures anchored in the 2 ways. Strain on the implant was investigated using a strain-gauged abutment, and load on the mucosa was measured using a suitably placed load cell. Ball attachments appeared to provide greater stability to the MIR-OVD, since load was more evenly distributed onto the distal mucosa of both sides. When the MIR-OVD was bar-anchored, axial load on the working-side abutment increased. PMID- 9857596 TI - Clinical and microbiologic effects of chemical versus mechanical cleansing in professional supportive implant therapy. AB - The aim of the present study was to compare the cleansing properties of mechanical supportive care for dental implants with the use of an etching gel. Sixteen patients underwent a 5-month clinical trial with monthly recalls. These patients, wearing maxillary complete dentures and mandibular overdentures supported by a bar device on 4 implants, were treated in a split-mouth study design. Test and control therapy were randomly assigned to left and right sides of the mandible. At the test side, 35% phosphoric etching gel (pH 1) was applied in the peri-implant sulcus. After 1 minute, the sulcus was thoroughly rinsed with a water spray for approximately 15 seconds per implants. Control therapy consisted of supra- and subgingival debridement using carbon fiber curettes and a rubber cup. Plaque, calculus, probing pocket depth, and modified Gingival Index were determined before each treatment. Microbiologic evaluation was performed at baseline, 1 month later, and 5 months later, just before and immediately after each treatment. Per treatment and per assessment, the mean scores of all clinical parameters were calculated for each patient. The number of colony-forming units was used as the primary efficacy variable in the analysis of microbiologic data. At baseline, no differences between test and control sites were observed for any of the clinical parameters. The mean Gingival Index and the mean probing pocket depth were reduced over the 5-month period. The mean reduction in Gingival Index at the test sites proved to be significantly larger at the control sites (P = .03). Both treatment modalities resulted in an instant reduction of the number of colony-forming units, where the reduction by chemical cleaning was larger (P < .05). This short-term study employing a high recall frequency indicates that local application of 35% phosphoric acid gel can be as effective as conventional mechanical supportive therapy. PMID- 9857598 TI - Soft liner-retained, implant-supported overdenture: a technical note. AB - This paper describes the design and fabrication of a soft liner-retained, implant supported overdenture used in 10 patients over a period of 1 to 6 years. A final complete denture impression is made of the dental arch, including implant abutments extending 5 mm above the gingival tissue. The master cast is fabricated with actual abutments placed in the impression. After the final wax try-in, the denture is flasked in the usual manner with the metal abutments in place. Before packing with acrylic resin, plastic tubing 1 to 2 mm thick is placed around each abutment. The denture is trial packed and allowed to bench cure over night. The plastic tubing is then removed and a soft denture lining material is placed in these areas only. The flask is closed and cured according to the manufacturer's recommendation. The denture is delivered as a conventional overdenture. PMID- 9857599 TI - Placement of dental implants without flap surgery: a clinical report. AB - Traditionally, the procedure of implant placement requires a surgical periosteal flap to be raised. In a percentage of implant cases, there is no need for flap surgery for implant placement, or for a follow-up surgical procedure for abutment connection. In this clinical investigation, 20 maxillary and mandibular implants were placed in seven adult male patients. The sites for implant placement were prepared according to an alternative surgical technique without raising a surgical flap. Patients were recalled periodically for 2 years to evaluate healing and clinical integration of implants. The results showed normal clinical healing at the first week of reexamination in all implant sites; periodontal probing of less than 2 mm circumferentially around all healing caps at 3 months and later at subsequent recall periods; no radiolucency observed in the peri implant zone; no sign of clinical mobility during recall examination; and no persistent or irreversible sign or symptoms of pain, infection, or necrosis. This alternative surgical technique can provide several advantages over the traditional 2-step procedure. PMID- 9857600 TI - Short-term healing following the use of calcium sulfate as a grafting material for sinus augmentation: a clinical report. AB - Because of the frequent lack of bone in the posterior maxilla, sinus augmentation has become a commonly practiced treatment modality. Many different materials have been used for augmenting the sinus, and the ideal graft is yet to be found. The present article reports the results of sinuses grafted with calcium sulfate in 2 patients. Bone biopsies were harvested 9 months after the augmentation procedure. In the first patient, 3 titanium threaded-cylinder implants were placed in the grafted area after 9 months, while in the second, 1 acid-etched, screw-shaped titanium implant was placed simultaneously with the graft. Light microscopic evaluation revealed new bone formation with ongoing remodeling and progressive lamellar maturation in the specimens. No remnants of the alloplastic material were detectable in any section, either within the bone or in the medullary tissue. When reevaluated at the uncovering procedure, the implants were radiographically and clinically judged to be osseointegrated. These observations suggest that, when used in the appropriate form and with the proper technique, calcium sulfate is a promising graft material for sinus augmentation, producing adequate quantity and quality of new bone for implant placement. PMID- 9857601 TI - Current knowledge in cleft lip and palate treatment from an orthodontist's point of view. AB - The aim of this review was to put new clinical research findings into proper perspectives relative to previously accepted knowledge on treatment of patients with cleft lip and palate. The first part of the paper deals with various aspects of infant orthopedic treatment, such as its influence on primary surgery, maxillary arch form and dimensions, feeding, psychological situation of the parents and speech development. Following parts analyze general maxillofacial growth outcome after surgery and also maxillofacial growth in relation to particular surgical procedures (palatal repair, periosteoplasty/gingivoplasty, bone grafting). The last part of the review discuss the effects of certain orthodontic/orthopedic treatment approaches as well as the role of dental implants in treatment of cleft lip and palate patients. PMID- 9857602 TI - Pain and discomfort experienced after placement of a conventional or a superelastic NiTi aligning archwire. A randomized clinical trial. AB - Two nickel-titanium arch-wire types commonly used for initial tooth alignment were compared with regard to the pain/discomfort patients experience during the initial phase of tooth movement. The two arch wires used were a superelastic nickel-titanium alloy, 0.014 inch Sentalloy, Light (GAC International Inc. Central Islip, NY, USA) and a 0.014 inch Nitinol (Unitek, Monrovia, CA, USA), a conventional nickel-titanium aligning archwire. One hundred and twenty-eight consecutive patients attending an orthodontic university clinic and 2 private practices for routine placement of a fixed appliance were randomly assigned one of these 2 initial arch wires. Assessments of pain/discomfort were made daily by means of a 100 mm visual analog scale (VAS) over the first 7-day period after bonding. On the first day, recordings were made every hour for the first 11 hours. The results showed that the level of discomfort increased continuously every hour after the insertion of either a Sentalloy or a Nitinol as first arch wires, with a peak in the first night, remaining high on the second day and decreasing thereafter to baseline level after 7 days. During the first 10 hours it was apparent that the pain/discomfort experienced after placement of a Sentalloy was less than that found with the Nitinol archwire, although a significant difference could be found at 4 hours only. No significant gender specific differences were found in either archwire group. A significant difference between the upper and lower dental arches was observed during the first 11 hours after placement of either a Sentalloy or a Nitinol arch wire, with the lower arch having the higher pain experience. PMID- 9857603 TI - The influence of the spring activator on the mobility of the lower jaw in traumatically injured patients. AB - For functional rehabilitation and improvement of mandibular mobility, 14 patients with dislocated collum or collum fracture dislocations were treated with a spring activator after surgical-conservative treatment. With an interincisal distance of < 12 mm, therapy was started with a loop spring. The average age of the patients was 26.6 years. Eight patients showed additional traumatic injuries, 2 patients were seriously polytraumatically injured. The clinical investigations referring to the maximum mouth opening and maximal interincisal distance resulted in a good to very good functional rehabilitation in all cases. The average maximal interincisal distance of 31.7 mm at the beginning of the spring activator therapy could be increased to 47.3 mm. The largest increase of the maximal interincisal distance was observed after 4 weeks of therapy. The type of reaction, however, differed interindividually. Three patients showed a spontaneous improvement of the interincisal distance within the first 2 to 3 weeks of removal of intermaxillary fixation, whereas 7 patients showed no improvement or only slightly improved values until insertion of the spring activator. An enlargement of the interincisal distance could be achieved in almost every patient within a relatively short period of time despite a drastically reduced mouth opening. Consequently, because of its mode of action in the form of a neuromuscular influence, the spring activator is especially suitable for the functional rehabilitation of patients with fractures of the condylar process. PMID- 9857604 TI - Treatment of Class II, Division 2 in the late growth period. AB - The "Deckbiss" with skeletal Class II jaw relationship sometimes presents a considerable therapeutic problem, particularly in the late growth period (DP3U), as regards the coordination of dental and skeletal treatment objectives. An effective treatment approach was demonstrated: a modified Herbst appliance used simultaneously with fixed appliances in the maxilla. The sample comprised 12 male (14.0 +/- 0.9 years old) and 10 female (12.3 +/- 0.4 years old) patients. Correction of the distal occlusion was achieved in all patients by means of the Herbst appliance, which was removed after an average time period of 6.4 +/- 0.2 months. In the mandible the multibracket appliances were then immediately inserted, and Class II elastics were used for retention. Maximum anchorage was required in the maxilla as well as in the mandible. Complete diagnostic records were made at the beginning of the treatment as well as 6 and 12 months later, in order to document skeletal and dental changes. A dental and skeletal Class I relationship was achieved in all cases. A significant improvement was recorded in the vertical jaw base relationship; this was still stable after a period of 12 months. In the dental area in particular, a so-called high-pull headgear effect (intrusion and distalization 16, 26) and intrusion of teeth 34, 44 were registered. Only a minor protrusion of the mandibular incisors was observed. Reinforcement of the bands reduced the failure rate significantly. The Herbst appliance does not represent a standard treatment for Class II. Its indication range is limited. PMID- 9857605 TI - In vitro evaluation of a measurement method to analyze the interdental, mesially directed force. AB - In order to evaluate interdental forces as a benchmark for mesial drift, a measurement technique was tested and evaluated on a human specimen. The measurement technique is based on the principle that a mesially directed horizontal force (FH) within a complete dental arch has an effect on interdental friction at the points of contact. The dynamic force (FZ), needed to pull out a defined metal strip from the interdental space is equal to the interdental frictional force (FR). Assuming unmodified approximal surfaces and unchanged tooth mobility in a complete dental arch, relative modifications of the interdental frictional force level provide a way of measuring horizontal force fluctuations. The validity of this measurement technique was measured by applying mesially directed forces of 1 to 5 N to the distal surface of the 2nd molar in a human specimen. The frictional forces measured increased in proportion to the distally applied force. The mesially directed force on the 2nd molar was transmitted in the dental arch anteriorly up to the incisor region and resulted in an increase of frictional forces. The reproducibility of this measurement technique was tested by quantitative analysis of potential measurement errors in the human specimen. The effect on interdental force measurement of the speed at which the metal strip is pulled was evaluated in a range of 50 to 500 mm/min. At the maximum pulling speed of 500 mm/min, a maximum scatter of 8% was recorded. Dependency of the dynamic force on direction of pull was measurable only when the metal strip was angled at more than 15 degrees. Experimental tests on the human specimen confirmed that the measurement technique presented here is sufficiently valid and reproducible for clinical long-term studies of interdental forces. PMID- 9857607 TI - Orthodontic treatment of a Class II/1 malocclusion with extremely enlarged overjet. AB - Orthodontic non-extraction treatment of a Class II malocclusion with an extremely enlarged overjet in a 9-year-old boy is demonstrated. PMID- 9857606 TI - The soft tissue cover of the mandibular condyle. Differentiation in histological forms and age-related changes of aggrecan- and versican-like proteoglycans. AB - Age-related changes of the composition of the extracellular matrix of the soft tissue cover of the mandibular condyle (STC), especially of the large proteoglycans, have been investigated. Proteoglycans were extracted from the STC of neonatal, juvenile and adult domestic pigs, fractionated by density gradient centrifugation and analyzed by electrophoresis/Western blotting. Experiments revealed firstly that a large CS/KS proteoglycan (aggrecan) is an essential constituent of the STC at all ages. This proteoglycan is required for nutrition of avascular tissues, and age-related changes in its average size and substitution with KS (keratan sulfate) may be a response to altered functional loading and tissue architecture of the STC. Secondly it was shown that a large CS/DS (chondroitin sulfate/dermatan sulfate) proteoglycan characterized by a doublet of core proteins at 200 and 250 kDa, thereby resembling perlecan, is present in the tissue of adults, but not of neonates and juveniles. Thirdly a large CS/DS proteoglycan characterized by core proteins at 350, 450 and 550 kDa, thereby resembling versican, was present in juveniles. It was detectable only weakly in neonates and not in adults. Results of core protein analysis were confirmed by results of agarose gel electrophoresis/Western blotting of the undigested proteoglycans isolated directly from the tissue extracts. Versican is believed to destabilize cell-matrix interactions required for cell proliferation and differentiation. In this context, presence of versican-like proteoglycans in the STC of growing individuals and its disappearance in adults appears to be related to the growth potential of the mandibular condyle. PMID- 9857608 TI - [Drug resistance in nosocomial strains of staphylococci to methicillin]. AB - The aim of the present study was the analysis of drug susceptibility of MRSA and MRCNS strains isolated from patients hospitalized in 14 wards of the State Clinical Hospital No 1 in Warsaw. The strains were identified (ID 32 STAPH), and their susceptibility to antimicrobial agents (ATB STAPH) was determined in ATB system (bioMerieux, France). Four methods were applied to confirm the resistance to methicillin: ATB-plus system, disc-diffusion method (Oxa 1 microgram, Oxoid, U.K.), Crystal MRSA ID (Becton Dickinson-BBL, USA) and agar screen test in TSA medium (Difco, USA) with methicillin (25 mg/l, Sigma, USA). 108 Staphylococcus spp. strains were found in 300 clinical specimens. 56 strains were methicillin resistant (52%). Among methicillin-resistant strains 13 MRSA, 28 MRSE and 15 of other species were found. All MRSA strains were susceptible to vancomycin, teicoplanin and fusidic acid. MRCNS were susceptible first of all to vancomycin (43/43), minocycline (42/43) and pristinamycin (42/43). On the basis of the obtained results it can be stated that methicillin-resistant staphylococci occur in hospital wards. The greatest number of methicillin-resistant strains was cultured from patients hospitalized in surgery wards (32), methicillin-resistant strains much more frequently occur among coagulase-negative staphylococci, especially in Staphylococcus epidermis. Glycopeptide antibiotics are most active against isolated MRSA strains. The most active therapeutic agent against MRCNS is vancomycin. PMID- 9857609 TI - [Utilization of siderophores from the Acinetobacter genus by staphylococcal bacilli]. AB - The ability of iron utilizing by means of siderophores produced by donor strains- the members of the genus Acinetobacter (8 strains) by 24 staphylococcal strains was investigated. All the donor strains synthesized hydroxamate class siderophores and six strains also catecholate class. The majority of staphylococcal strains could utilize these siderophores. Most strains utilized siderophores from A. juni 321 and A. johnsonii 349 strains. Only three staphylococcal strains were not be able to utilize siderophores from all donor strains. PMID- 9857610 TI - [Examination of selected biological properties for swarm cells in Proteus mirabilis]. AB - Certain biological properties, which generally are thought to play a role in the bacterial pathogenicity, were compared in short rods, swarm cells and penicillin induced filaments of Proteus mirabilis S1959. Swarm cells of P. mirabilis S1959 weakly adhere to human uroepithelial cells and also do not penetrate L929 line of mouse fibroblasts. They do not show any cytotoxic activity, are poorly phagocytized by macrophages and granulocytes and start to divide by the end of the phagocytosis process. Their well-marked cell-bound haemolytic activity is correlated with fast division to the short rods. In in vivo experiments we have demonstrated the presence of long filamentous multinucleate nonseptate cells in the bladder and in the kidneys of infected animals. However there was no correlation between the number of swarm cells seen under microscopic examination and the intensity of infection. PMID- 9857611 TI - [Occurrence of extended spectrum beta-lactamases (ESBL) and inducible beta lactamases (IBL) in clinical strains of gram-negative rods]. AB - This study was undertaken to check the situation concerning the occurrence of Gram-negative rods producing extended-spectrum beta-lactamases (ESBL) and inducible beta-lactamases (IBL) in clinical specimens from patients hospitalized in National Clinical Hospital No. 1 in Warsaw. Such determinations were not performed in this hospital so far. During three months (April-June, 1997) 200 strains of Gram-negative rods were cultured. The strains were identified in automatic ATB system using strips with biochemical tests: ID 32 E for enteric rods and ID 32 GN for non-fermenting rods. ESBL-producing strains were detected with double disc diffusion test according to Jarlier et al. (1988). Clavulanate was applied as the inhibitor of beta-lactamases (AMO/CLAV disc). Inducible beta lactamases were determined using double disc method according to Sanders and Sanders (1979). Cefoxitin was the inductor of these beta-lactamases. 82 strains (41% of all strains) belonging to Enterobacteriaceae family, 92 strains (46%) of Pseudomonadaceae rods and 26 strains (13%) of other Gram-negative rods were isolated. 30 ESBL-producing strains (15% of all strains) and 45 strains (22.5%) with IBL activity were detected. The obtained results confirm the necessity of continuous and reliable monitoring of ESBL--and IBL--producing strains among Gram negative rods isolated from clinical materials. The aims of such procedure are the control and prevention of their dissemination within a hospital as well as the avoidance of therapeutic failures. PMID- 9857612 TI - [Evaluation of in vitro susceptibility of hospital bacterial isolates to piperacillin and tazocin (piperacillin/tazobactam]. AB - Due to increasing frequency of infections caused by pathogens that are resistant to beta-lactam antibiotics, combinations of such antibiotics and beta-lactamase inhibitors were introduced into therapy in last few years. Tazobactam is the most potent beta-lactamase inhibitor. The purpose of the study was to evaluate in vitro susceptibility to piperacillin and piperacillin with tazobactam of 256 isolates cultured from biological samples obtained from 203 patients. The biological materials obtained were as follows: urine (44.9%), post-operative and post-traumatic wound swabs (27.3%), BAL (12.1%), blood (6.6%), drain swabs and other (5.5%). The isolates predominantly found were Escherichia coli (22.3%), Pseudomonas aeruginosa (16.0%), Staphylococcus aureus MSSA (13.7%), Proteus mirabilis (11.7%) and other. There were 95.5% of strains found susceptible to piperacillin with tazobactam and only 4.3% resistant ones. On the other hand, piperacillin only susceptible strains were 59.4% and resistant ones in 40.6%. Great differences in susceptibility to examined antimicrobial agents were observed in Enterobacteriaceae family and Staphylococcus (MSS) genus. There were no differences in susceptibility to piperacillin and tazobactam and piperacillin alone in anaerobic Enterobacteriaceae strains and non-fermenting Gram-negative bacilli. PMID- 9857613 TI - [Examination of protein MPB 70 properties in BCG substrains used for the production of Polish antituberculosis vaccines]. AB - The aim of the study was examination of MPB70 protein production by different BCG substrains and testing this protein as reagent in allergenic skin test on BCG vaccinated guinea pigs. Three BCG substrains: Danish 1331 (D), Japanese 172 (J) and Polish BCG Moreau (P) were used for the study. The protein MPB70 received from dr Nagai from Osaka University was used as reference preparation. It has been shown that MPB70 protein of molecular weight about 21,500 Da was present only in BCG Moreau (P) and Japanese (J) substrains. In immunoblotting test monoclonal antibodies reacted additionally with 43,000 Da protein in all tested substrains. We suspect that the detected additional molecule was a dimer of MPB70 protein. The development of skin reactions to MPB70 protein was seen in guinea pigs vaccinated with J and P substrains. The peak of alergy to MPB70 protein was observed 9 weeks after vaccination. In sera of vaccinated animals antibodies against MPB70 protein were detected by ELISA method. The possibility of using MPB70 protein for diagnostic purpose is discussed. PMID- 9857614 TI - [Detection of toxin producing strains of Clostridium difficile using rapid diagnostic methods]. AB - Feces of 53 patients from different hospital wards suffering from long term post antibiotic therapy diarrhea were tested. For direct detection of C. difficile toxin A, in samples TCD (Becton-Dickinson), and C. difficile Toxin A Test (Oxoid) tests were used. Toxin A was detected in 16 samples (29.6% tested). C. difficile strains were isolated from 40% of the fecal samples. Toxin A was detected in 25 Clostridium difficile strains with commercial tests and toxin B was detected using McCoy cell line. Toxin A was not produced by 3 C. difficile strains in vitro, but toxin B was produced by all strains. The polymerase chain reaction (PCR) test showed that all isolated strains possess genes of toxins A and B. PMID- 9857615 TI - [Bacterial flora in pharyngitis and tonsillitis]. AB - The aim of the study was a microbiological analysis of pharyngeal swabs obtained from 158 patients with the diagnosis of pharyngitis and purulent exudates from the tonsillar crypts of 10 patients treated for chronic purulent tonsillitis. Beta haemolytic streptococci groups A, B, C and G were isolated from 30% of the patients. The most frequently isolated were Streptococcus pyogenes--12% of patients and Streptococcus group C--10.7%. Other streptococci were isolated less frequently: Streptococcus group B--44%, group G--2.5%. The majority of isolated bacteria belonged to potential pathogenic flora (70% patients). Staphylococcus aureus (37%) and Haemophilus spp. (36%) were isolated most frequently. Other bacteria were isolated in the following sequence: Moraxella catarrhalis--22%, Streptococcus pneumoniae--17% and Gram-negative rods from the Enterobacteriaceae family--6%. One case of Plaut-Vincent tonsillitis was diagnosed. Aerobic and anaerobic bacteria were isolated from purulent exudates from the tonsillar crypts of 10 patients treated for chronic purulent tonsillitis. The isolated anaerobic bacteria belonged to genus of Prevotella, Fusobacterium, Peptostreptococcus and Gemella. PMID- 9857616 TI - [Susceptibility of clinical strains of gram-positive bacteria to selected beta lactam antibiotics]. AB - The aim of the study was to determine the activity of four beta-lactam antibiotics against nosocomial strains of Gram-positive bacteria. Two antibiotics combined with beta-lactamase inhibitors: timentin (TIC/CLAV) and tazocin (PIP/TZB) and two carbapenems: imipenem and meropenem were applied. The clinical strains were isolated from patients hospitalized in surgical ward of the National Clinical Hospital No 1 in Warsaw. The strains were identified in the automatic ATB system using ID 32 STAPH, API STREP, API CORYNE and API 20 A strips. The susceptibility of isolates to antibacterial agents was determined in the automatic ATB system using ATB STAPH, ATB STREP and ATB ANA strips. The susceptibility of strains to timentin, tazocin, imipenem and meropenem was tested with disc diffusion method. 111 strains of Gram-positive bacteria were cultured. Staphylococci (49) and enterococci (44) dominated among isolated strains. 33 Staphylococcus spp. strains were identified as methicillin-resistant. The obtained results indicate a significant role of Gram-positive cocci (staphylococci and enterococci) in the aetiology of nosocomial infections. Antibiotics combined with beta-lactamase inhibitors and carbapenems demonstrate broad antibacterial spectrum against clinical strains of Gram-positive bacteria except E. faecium strains. PMID- 9857617 TI - [The role of rotaviruses in digestive tract infections of hospitalized children with diarrhoea at the Health Care Consortium in Sokol Podlaski]. AB - The aim of the study was a trial of establishing of the frequency of rotavirus infection and mixed bacterial-viral infections in children treated for diarrhoea in a hospital in the period from July 1 1996 to June 30 1997. Moreover, an attempt was made at establishing of the frequency of rotavirus colonization of the digestive tract in healthy newborns born in that hospital during the period of that study. The studies were done on feces samples from 169 children with diarrhoea from 4 days to 13 years, and from 30 healthy peers. In all samples rotaviruses were sought along with pathogenic Enterobacteriaceae Tests for rotaviruses were done with Slidex-Rota Kit 2 (Bio-Merieux) and for EPEC with the EPEC latex test (Biomex). In all, rotaviruses were found in 88 ill children (52.1%) including mixed bacterial-viral infections in 10 (5.1%) children. In 9 children beside rotaviruses enteropathogenic E. coli (EPEC) were disclosed, and in 1 child S. enteritidis. Moreover, in 1 child (0.6%) culture yielded simultaneously S. sonnei and E. coli 0127. No rotaviruses were found in any 30 healthy newborns, but from one of them E. coli 018 was cultured. The second most frequent aetiological factor in diarrhoea were EPEC organisms found in 17 (10.0%) children, the third factor were S. enteritidis strains in 6 children (3.6%). In one case S. sonnei and E. coli 0127 were obtained from faces. Rotavirus infection was most frequent in children aged from 2 months to 3 years, and EPEC infection in children up to 2 years. The incidence was highest in winter/spring, with peak in April when 27 cases of rotavirus diarrhoea were noted. PMID- 9857618 TI - [Evaluation of the usefulness of Bordetella pertussis toxins for serodiagnosis of whooping cough]. AB - The results of serological tests for the detection of antibodies against pertussis toxin by means of the ELISA test, and endotoxin of B. pertussis in the passive haemagglutination test were analysed. The levels of IgG, IgM and IgA antibodies were determined for 95% of serum samples of 108 children in control group (without evidence of respiratory infections) for establishing of the correlation of these with the age of these children. In the light of these data the results were evaluated of testing of 136 serum samples taken from 127 children with suspected whooping-cough. The highest diagnostic importance had the demonstration of high IgA or/and IgG immunoglobulins level against pertussis toxin in ELISA test, or tracing of the rate of rise of antipertussis antibodies in the passive haemagglutination test with B. pertussis endotoxin in children with clinical symptoms of whooping-cough. PMID- 9857619 TI - [The presence of antibodies to Borrelia burgdorferi associated with immunologic complexes in sera of foresters]. AB - Binding of antibodies specific to Borrelia burgdorferi in circulating immune complexes can lead to false negative results in serological tests. The aim of our study was to determine the presence of IgM antibodies to Borrelia burgdorferi bound in immune complexes in 52 sera of foresters the National Park in Karkonosze. Free and bound in immune complexes IgM antibodies present in 6 (11.5%) examined sera. In 24 (46.2%) seronegative sera after dissociation of immune complexes IgM antibodies to spirochaeta were found. The rest of the examined seronegative sera we failed to find IgM antibodies to Borrelia burgdorferi. The diagnostic assay, such as antibody analysis of immune components is useful in establishing of the diagnosis of borreliosis in seronegative cases and monitoring of disease activity. That method should be introduced for routine diagnosis of Lyme disease. PMID- 9857620 TI - [Distribution of IgG subclasses and their biological activity in bioglobulin]. AB - Preparations of human immunoglobulin for intravenous use (IVIG) should contain proper percentage of IgG subclasses, corresponding to physiological proportions with preserved activity of antibodies. The amounts of IgG subclasses were determined using the radial immunodiffusion technique against to the WHO reference serum 67/97. It appears that variations are observed among the different lots and manufacturers of the various formulations of IVIG available in Poland. Although some of these differences are not statistically significant, significant differences in subclass concentrations were found between formulation of IVIG, with Bioglobulina being lower in IgG1 and higher in IgG2 and Endobulin having a lower per cent of IgG3. The study was aimed at establishment of activity values against diphtheria and tetanus toxoids and Streptococcus group B (GBS). IgG antibodies to tetanus and diphtheria toxoid antigens were predominantly of the IgG1 and IgG2 isotype. IgG4 subclass, was present in smaller proportion and IgG3 fractions was absent. Antibodies to GBS were only IgG2 and IgG1 isotypes. Basing on obtained results it can be stated that in spite of the imbalance between the amounts of IgG1 and IgG2 in Bioglobulina, the activity which was determined in subclasses is comparable with that of other IVIGs. PMID- 9857621 TI - [Detection of bacterial biofilm on medical biomaterials]. AB - This study was performed to assess the value TTC assay in the diagnosis of biomaterial-associated infections. In this assay, soluble colourless TTC is reduced to insoluble red formazan by electron transfer associated with active oxidative bacterial metabolism and is precipitated intracellularly. Microbial adhesion and biofilm formation on the surface of medical prosthetic devices (vesicular and urinary catheters) made of various polymers (PTFE, H-PE, PCW, SL), were determined. The microorganisms which are most often isolated in medical device-associated infections: S. aureus, S. epidermidis, E. faecalis, E. coli, P. vulgaris, P. aeruginosa, C. albicans, were included into the study. The obtained results indicate that the assay using TTC as a metabolic indicator of bacterial biofilm presence, is technically simple to conduct with minimal setup time. Even when classical cultures yielded no bacterial growth, TTC assessments demonstrated bacterial biofilms. TTC assay could be recommended as a quick routine method for confirmation of biomaterial device-associated infection. PMID- 9857622 TI - [A new method for evaluation of the antagonistic action of bacterial lactic acid (LAB) on selected pathogenic indicator bacteria]. AB - A comparison was made of three methods: paper disc, double layer and a newly proposed agar slab techniques for testing antibacterial activity of Lactobacillus strains. Strains of indicator bacteria were selected from important pathogens of the human alimentary and genitourinary tract. The agar slab method, which is based on applying of slabs of MRS agar with overnight growth of the antagonistic bacteria to the surface of appropriate agar plates seeded with indicator bacteria, appeared to be the only method suitable for testing of aerobic and anaerobic fastidious bacteria. After testing of 27 Lactobacillus strains against 3 selected indicator bacteria: E. coli, Salmonella enteritidis and Staphylococcus aureus, calculating mean values and standard deviations for diameters of inhibitory zones and making variance analysis it was shown that in comparison to other methods the agar slab technique, although less sensitive than others, gave most consistent and reproducible results. PMID- 9857623 TI - [First page]. PMID- 9857624 TI - [Unreliability of pulse oximetry in hypothermic children after cardiovascular surgery with deep hypothermic circulation]. AB - BACKGROUND: The aim of the study was to evaluate the accuracy of oxygen saturation with the pulse oximeter (SpO2) in children undergoing cardiovascular surgery with deep hypothermic circulation. METHODS: The SpO2 values measured at the hand and at the foot were compared in 50 patients with the arterial oxygen saturation (SaO2) measured with the oximeter. A variance value between the two measurements less than 3% was considered as a reliable measurement, and the precision of the measurement was defined as the standard deviation of the variability. RESULTS: For skin temperature between 35 and 28 C grade, and for core temperature more than 34 grade, the SpO2 is a reliable measurement of the SaO2; for skin temperature less than 28 grade and core temperature less than 34 grade the SpO2 do not correspond the SaO2. CONCLUSIONS: The variability and the precision of the measurements are not affected the position of the probe (hand or foot). PMID- 9857625 TI - [The use of hydrophobic PALL BB22-15MS filters in cross-infection prevention during anesthesia]. AB - OBJECTIVE: Microbiological testing evaluations of hygienic procedure properties using PALL BB22-15MS filters to prevent contamination of "single use" breathing circuits during anesthesia. DESIGN: Prospective study. SETTING: Two operating rooms in a University hospital. PATIENTS: One hundred and thirty eight patients underwent general anesthesia for urologic surgery procedures. Patients with positive tests for HIV, B and C hepatitis and those considered to be at risk for HIV infection were excluded. The study was divided into five phases on the basis of times of usage of the same circuit for an increasing number of patients: in phase I, microbiological tests were performed on 4 circuits used on 4 different patients; in phase II the same tests were performed on 2 circuits each used on two different groups of three patients; in phase III, a circuit was used on a group of 15 patients and another on a group of 16 patients and the results were analysed; in phase IV a circuit used in a group of 32 patients was evaluated and phase V involved the analysis of a circuit that was used in a group of 65 patients. INTERVENTIONS: A filter was left in place between the patient and the circuit's Y-piece during all phases of anesthesia. The level of microbial contamination of breathing circuits was analysed in order to evaluate the reliability of the procedure. RESULTS: All analysed circuits remained uncontaminated. Staphylococcus hominis was revealed in respiratory circuit no. 6 of phase II, probably as a consequence of secondary contamination. CONCLUSIONS: Our results confirm that this procedure with the routine placement of a PALL BB22 15MS filter at the circuit's Y-piece can provide an adequate level of protection against cross-infections during anesthesia. The hygienic protocol proposed may allow the change of the anesthetic breathing only once a month. PMID- 9857626 TI - [Rapidity of emergence from anesthesia with sevoflurane or isoflurane]. AB - BACKGROUND: Sevoflurane, a new anesthetic agent recently introduced in Italy, is characterized by a low coefficient of distribution blood/gas that is associated both quick narcosis and fast emergence from anesthesia. Aim of this study is the comparison between two different anesthetic methods in patients submitted to general, gynecological and orthopedical surgical operations. METHODS: Thirty subjects randomized in two groups of fifteen patients each have been studied. In the first group (8 females, 7 males), sevoflurane as maintenance anesthetic was utilized; in the second group (5 females, 10 males), isoflurane was used. The final course of anesthesia were the same for both groups. ECG, FC, NIBP, FiO2, Paw, Spirometry, ETCO2, Fi-Alogenates, ET-Alogenates, MAC were monitored in all patients. In each group of patients the emergence time, extubation time, and dismission from operatory room were compared. Cardiocirculatory performance was evaluated monitoring arterial pressure and cardiac frequency in the same time of intubation and every 5 minutes following until extubation time. RESULTS: The incidence of some postoperatory collateral effects was recorded too. The three emergence times showed a significative difference in favour of patients submitted to sevoflurane narcosis. Both groups of patients showed also appearance of nausea, vomiting and shived in similar way. CONCLUSIONS: On the basis of the results obtained, the conclusion is drawn that sevoflurane is more rapidly eliminated than isoflurane, but both give a good cardiocirculatory performance. PMID- 9857627 TI - Experimental and clinical studies about the preemptive analgesia with local anesthetics. Possible reasons of the failure. AB - Though we fully agree with the truthfulness of the physiopathological sequences proving the phenomena of the hyperalgesia onset and therefore a prolongation of the postoperative pain, we do not believe, as stated by some authors that the studies carried out up to now, both experimental and clinical, are sufficient, the first ones to confirm the preemptive analgesia, while the second ones to be defined as contrasting and totally inadequate to confirm the preemptive analgesia effect. The lack of positive clinical data is why some authors have suggested a "revision" and a "reduction" of the word preemptive analgesia. On the contrary, we believe that differences which seem to originate from the clinical works can also be found in the experimental ones if we examine them with the same methodological principle. In order to classify better such an important treatment like the preemptive analgesia, the aim of this note is to focus the attention on the experimental and clinical data. Our criterion to select the works has based on a backwards research taking into consideration the last reviews and studies about the preemptive analgesia. Since the results obtained with local anesthetics or with opioids seem to counterbalance each other, we have reexamined the experimental and clinical works limiting our attention to the local anesthetics. On a total of 19 assessments carried out in 16 experimental studies, only three of the eight ones comparing pre-vs post-administration prove a real preemptive analgesia effect of the local anesthetic (block of the first and second phase of hyperexcitability) whereas four of the eleven clinical studies examined seem to confirm the validity of the preemptive analgesia effect. These data show how the results of the experimental studies are equal to the results of the clinical ones. However, an important characteristic that seems to come out from these works is connected to two elements strictly linked each other: the intensity and the duration of the nociceptive impulse produced and the level and the type of the induced block. In our opinion, the failure of many of these works can be attributed to the inadequacy of the analgesic levels (intensity of the block) reached and maintained in the pre-and intraoperative period. Therefore we believe that before thinking of a "review" and "reduction" of the word preemptive analgesia is necessary to take a step backward and to face again the methodological problems of the preemptive analgesia. PMID- 9857628 TI - [Cardiopulmonary resuscitation. The effect of severe hypothermia on clinical outcome]. AB - BACKGROUND: Evaluation of outcome after CPR in severe hypothermic patients. DESIGN: Perspective study from October 1995 to April 1996. SETTING: First aid team of Italian Red Cross, Busto Arsizio (Varese), Italy. METHODS: A population of 22 patients in cardiac arrest in which CPR was performed immediately after rescue team's arrival is studied. ECG, core temperature, SpO2 and MAP were monitored whereas vital parameters were present during Basic Life Support. Outcome after CPR was evaluated with GOS scale. RESULTS: It has been observed that severe hypothermia and time of cardiac arrest impact on the clinical outcome after CPR. The high mortality rate after CPR with BLS standard is worsened by a core temperature < or = 33 degrees C. CONCLUSIONS: Severe hypothermia seems to have a dangerous effect upon outcome after cardiopulmonary resuscitation; heating systems for body temperature could prevent this situation improving CPR results. PMID- 9857629 TI - [Leptospirosis. Description of a clinical case and review of the literature]. AB - Leptospirosis is a world-wide diffused anthropozoonosis due to many strains of Leptospira. Initial symptoms may be mild, although in many cases severe systemic symptoms, including high fever, hypotension, etc. may be present since the beginning. In these latter circumstances, the diagnosis of leptospirosis can be very difficult because of the complexity of clinical picture especially when the history is lacking or incomplete. A case report of a 45 year-old man admitted to the hospital after severe jaundice and fever of unknown origin associated to altered mental status, renal failure and hypoxemia is presented. Because of the presence of septic shock and severe respiratory failure, the patient was transferred to the intensive care unit. The diagnostic hypothesis, based on clinical history, was confirmed by laboratory tests (leptospiral IgM antibodies detection). Therapeutical approach with the use of selected antibiotics (penicillin 24,000,000 U for day) and therapy of septic shock led to improvement of the patient's clinical conditions who was then transferred to a regular medical ward. PMID- 9857630 TI - Severe accidental hypothermia successfully treated by warmed peritoneal lavage. AB - A case of severe accidental hypothermia (core temperature 20 degrees C) in an elderly woman successfully treated by warmed lavage of peritoneal cavity is reported. The various available rewarming techniques and some observations about the differential diagnosis between the severe hypothermic patient with cardiac arrest and the hypothermic dead person are briefly reviewed on the basis of current literature. PMID- 9857631 TI - The educational activities of the European Academy of Anaesthesiology. AB - The European Academy of Anaesthesiology was founded in 1978 at the time of the introduction of the Medical Directives of the European Union. This article reviews the development of some of the educational work of the Academy including the foundation of its journal, the European Journal of Anaesthesiology. The article also considers in some details the development of the European Diploma in Anaesthesiology and Intensive Care since its inception in 1984 to the present time. PMID- 9857632 TI - Births, marriages, divorces, and deaths: provisional data for July 1998. PMID- 9857633 TI - [Correlation between semi-quantitative classification of corneal topography and the coefficient of Fourier transformation after non-mechanical perforating keratoplasty]. AB - BACKGROUND: Semiquantitative classification of corneal topography after penetrating keratoplasty has the potential for focusing information about the areal dioptric power of the cornea. The purpose of this study was to objectify the procedure of manual semiquantitative classification using a Fourier transform of corneal topography power data and to correlate both methods. PATIENTS AND METHODS: Fifty patients each (30 keratoconus, 20 Fuchs dystrophy) underwent nonmechanical trephination (excimer laser MEL60, Aesculap-Meditec, Jena) in penetrating keratoplasty. All procedures (7.5-mm trephination diameter in Fuchs, 8.0 mm in keratoconus, double-running 10-0 nylon suture) were done by one surgeon. Pre-, intra- and postoperative treatment were identical. At the follow up examinations, the keratometric astigmatism, qualitative and quantitative criteria of the automatic videokeratography, visual acuity and refraction were assessed. Corneal topography was classified both manually and based on Fourier coefficients. RESULTS: After a mean follow-up of 24 +/- 5 months, keratometric net astigmatism was 3.0 and 2.7 D with keratoconus and Fuchs dystrophy. Corneal topography analysis showed a higher orthogonality of the bow-tie shape and less asymmetry between opposite hemimeridians with increasing follow-up after keratoplasty. The semiquantitative classification showed a statistically significant correlation with the classification based on Fourier coefficients, especially with higher astigmatism and after suture removal (P = 0.04/0.01 before/after suture removal). DISCUSSION: After nonmechanical trephination, the semiquantitative classification of corneal topography can be synthetized using Fourier analysis of corneal dioptric power data. In the future, this method may be favored for prediction of potential best-corrected visual acuity after penetrating keratoplasty. PMID- 9857634 TI - [Depth of lamellar keratoplasty with the guided trephine system for transplantation of full-thickness donor sections]. AB - BACKGROUND: Despite the fact that deep lamellar keratoplasty (DLKP) is less invasive than to penetrating keratoplasty (PKP), this procedure is rarely performed. We therefore investigated whether or not the DLKP technique we employed can achieve stable improvement of visual acuity. MATERIALS AND METHODS: Thirty-three eyes underwent TLKP for treatment of superficial corneal pathology. The donor tissue transplanted was suitable for PKP. The donor lenticule was obtained on the artificial chamber of the guided trephine system (GTS). The recipient cornea was trephined with the same trephine to a depth of 680 microns. Manual dissection was performed with a bevel-up blade. The donor lenticule with the endothelium peeled off was then sutured in with a 10 x 0 nylon double-running antitorque suture. Cortisone-antibiotic eye drops were administered postoperatively. RESULTS: Throughout the series no complications occurred. The mean best corrected visual acuity (BSCVA) over glasses was 0.29 (+/- 0.21) preoperatively, 0.1 (+/- 0.11) at 1 week, 0.33 (+/- 0.14) at 1 month, 0.5 (+/- 0.13) at 6 months, 0.61 (+/- 0.16) at 1 year and 0.63 (+/- 0.15) at 2 years. Clinically, we observed two subpopulations. In the first group of 87% of the cases, mean BSCVA was 0.67 (+/- 0.07) at 6 months. The remaining cases (BSCVA < or = 0.25 at 6 months) achieved a mean BSCVA of only 0.2 (+/- 0.04) at 1 year. Mean corneal astigmatism measured 2.93 D (+/- 1.62) preoperatively, 2.69 D (+/- 1.18) at 1 month, 2.09 D (+/- 1.07) at 1 year, and 2.22 D (+/- 1.11) at 2 years. We did not observe any graft rejection. CONCLUSION: The technique reported for DLKP provides excellent matching of donor lenticule and recipient bed. Separation of donor and recipient stroma prevents interface healing. DLKP appears to be a safe procedure for the surgical treatment of superficial corneal pathology and may offer a clinically applicable alternative to PKP. PMID- 9857635 TI - [Effect of E. coli endotoxin on auto-/paracrine function and endothelial cell loss of donor corneas in organ culture]. AB - Corneal cells are known to participate in the regulation of local inflammatory processes by secretion of cytokines. As the corneal endothelium may be exposed to endotoxin in organ culture and endotoxin is known to trigger inflammatory reactions, we investigated the effect of endotoxin from E. coli on organ cultured donor corneae with respect to autocrine and paracrine functions and the endothelial viability and density. 6 pairs of donor corneae were transferred to organ culture. Medium samples were taken prospectively from day 0 to day 20. On day 10 the medium was changed and one of each pair was incubated with 50 micrograms/ml of endotoxin while the other was immersed in standard organ culture medium. The samples were screened for IL-1, -2, -4, -5, -6, -8, -10, TNF alpha and GM-CSF by ELISA. In addition endothelial cell counts were performed at day 0, after 10 and after 20 days of organ culture, using the fixed frame technique. All endotoxin-incubated organ culture media showed significantly increased IL-6 and 8 levels compared to the fellow cornea and to pre-exposure levels (P < or = 0.004). In the endotoxin-treated corneae a significantly higher endothelial cell loss occurred (P = 0.007) and signs of degeneration were observed. None of the other cytokines (IL-1 alpha, IL-2, IL-4, IL-5, IL-10, GM-CSF and TNF alpha) were detected during either the first (1 to 10-day) or the second (11 to 20-day) phase of incubation. A correlation seems to exist between an increase in IL-6 and -8 induced by endotoxin, and endothelial cell loss in organ culture. PMID- 9857636 TI - [Radiotherapy of exudative senile macular degeneration. A prospective controlled study]. AB - Despite the successful therapy of subretinal neovascular membranes by laserphotocoagulation there are many problems to be overcome. In the case of subfoveolar neovascularization, photocoagulation leads to a sudden decrease in visual acuity. Recently radiotherapy is considered as an alternative. Complications and effectivity were evaluated in this prospective and randomized trial. The initial results are presented. PATIENTS AND METHODS: There are 76 patients (51 women, 25 men, average age 77.7 +/- 8.6 years) included in the prospective randomized study. All of them show subfoveolar neovascular membranes in FLA and a decrease in visual acuity between 0.05 and 0.5. They were randomly assigned to either the radiotherapy or the control group. Radiotherapy was done within 6 days by 6 x 2 Gy (6 MV photons). The follow-up was at 4 weeks, after 3 months, after 6 months and then every 6 months after the end of radiotherapy. On average the follow-up is at 15.1 months. RESULTS: Concerning age and visual acuity before therapy, the control group and the radiotherapy group were not significantly different. At 4 weeks after radiotherapy, visual acuity was 0.13 +/ 0.46 (LogMAR). After 12 months, visual acuity at a distance was 0.11 +/- 0.30 in the therapy group and 0.09 +/- 0.13 (P = 0.838) in the control group. Patients with a preoperative visual acuity better than 0.2 improved more after radiotherapy. Metamorphopsy improved in 75% of the therapy group. The following complications could be observed: In the control group 3 patients suffered subretinal bleeding, in the radiotherapy group 3 patients, respectively. CONCLUSIONS: At present, the follow-up is too short to recommend radiotherapy as a standard procedure in the case of subfoveolar neovascularization. The results in patients with a better preoperative visual acuity encourage us to continue this study. PMID- 9857637 TI - [Trans-pupillary hyperthermia of malignant choroid melanoma. Initial experiences]. AB - The purpose of this report is to evaluate the efficacy of transpupillary thermotherapy (TTT) to treat choroidal melanoma. PATIENTS AND METHODS: Twelve patients with choroidal melanoma were treated. Four with small tumors close to the posterior pole received TTT alone; 8 were treated simultaneously with TTT and Ru-106 brachytherapy. To perform TTT a diode laser with a beam diameter of 3 mm was used. Exposure time ranged from 60 to 90 s until the retinal surface became light gray. Follow-up examinations were performed at 1-month intervals. If the tumor was partially regressed, additional TTT was performed to reach the endpoint of a chorioretinal scar. RESULTS: All tumors exhibited a reduction of tumor height in a maximal follow-up period of 6 months. Four cases showed complete regression within 3 months. Side effects were minimal. CONCLUSIONS: Treatment with TTT may be useful as a complementary modality to brachytherapy. A longer follow-up time is necessary for final evaluation. PMID- 9857638 TI - [Treatment of peri-ocular skin lesions with the erbium:YAG laser]. AB - INTRODUCTION: A great number of consmetically disturbing, cutaneous lesions are localized in the periocular region. While various approaches for treatment such as excision, electrocauterization or cryosurgery often show unsatisfactory results, the use of laser technology is of increasing interest. MATERIAL AND METHODS: A total of 104 patients with different cutaneous periorbital lesions (wrinkles, xanthelasma, syringoma) were treated with a new erbium: YAG laser system (wavelength 2940 nm, pulse duration 0.350 ms and pulse energy 0.1-1.7 Joules), which works on the principle of vaporization. RESULTS: The erbium: YAG laser allows athermic ablation of very thin skin layers without scarring and with minimal lateral thermal injury due an extremely short pulse duration. Periorbital lesions can be treated effectively by erbium: YAG laser, and good to excellent cosmetic results can be obtained. CONCLUSIONS: The use of pulsed erbium: YAG laser is an effective and promising new method for treatment of different superficial periorbital cutaneous lesions. PMID- 9857639 TI - [New European medical flight safety guidelines. Their significance for the ophthalmologists]. AB - In 1996 the new European Flight Crew Medical Requirements were finalized. The ophthalmological changes compared to the existing German standards range from the organization of evaluations to the necessary frequency of examinations. They also cover different examination techniques and different criteria for fitness. The changes in the requirements for ophthalmological fitness primarily affect ametropia, anisometropia, color vision and phoria. Knowledge of the European requirements is very important to advise pilots and any patient interested in becoming a pilot. PMID- 9857640 TI - [Low Vision Enhancement System (LVES). Initial clinical experiences with a new kind of optoelectronic rehabilitation system]. AB - BACKGROUND: The Low-Vision-Enhancement-System (LVES) is the first binocular optoelectronic rehabilitation device with variable focus distance. METHOD: LVES was attempted on 25 patients who were not adequately treatable with classic rehabilitation devices. Using the new device, short-distance vision as well as the ability to read and write were tested. RESULTS: In six patients, a significant improvement was achieved using LVES. In these patients, short distance vision as measured by LVES varied from 0.63 to 1.0. All succeeded in reading fluently, and five patients, in writing. Of these patients, five suffered from large central scotomas due to macular disease, and one female patient, from post-traumatic optic nerve atrophy. In 10 patients, near-sighted vision could be improved although the ability to read and write could not be restored, while nine patients did not show any benefit. In the latter group, were four patients with retinopathia pigmentosa, who complained of the further reduction in visual field due to the system. CONCLUSION: LVES is an additional device for the rehabilitation of low-vision patients which is especially useful in cases of macular diseases. PMID- 9857641 TI - [Recurrent panuveitis. First manifestation of Behcet disease in childhood]. AB - BACKGROUND: Behcet's disease is a rare systemic vasculitis of unknown etiology. The typical symptoms include recurrent oral and/or genital aphthous lesions, iridocyclitis (historically with hypopyon) and various skin lessions. The number of young adults and children which are diagnosed with Behcet-syndrome is increasing in recent years and the mean age of manifestation has decreased to 25 years in German patients. The disease is generally diagnosed later in German (48.5 months) than in Turkish patients (25.5 months). Ocular manifestation has been confirmed as a marker of severe prognosis. In 15-25% of affected patients it leads to blindness. PATIENT: We describe a 19-year-old Caucasian woman who has suffered from the typical symptoms (oral aphthous lesions, recurrent uveitis posterior, various skin lesions) since the 16th year of age. RESULTS: After three years the patient was finally be diagnosed with Behcet's disease (using the criteria of the "International Study Group for Behcet's Disease"). Visual acuity was stabilized with an immunosuppressive therapy, although there was no complete remission in disease activity. In addition a cerebral vasculitis was manifested. CONCLUSIONS: Ophthalmologists should be familiar with Behcet's disease. In cases of recurrent uveitis Behcet's disease should be included in the differential diagnosis because timely immunosuppressive therapy can prevent irrevocable changes in the corpus vitreum and retina and preserve complete visual acuity. PMID- 9857642 TI - [Eye involvement in porphyria erythropoetica congenita]. PMID- 9857643 TI - [Immunosuppressive therapy of intraocular inflammation. I. Basic principles, indications, contraindications, side-effects]. PMID- 9857644 TI - [Repeated paracentesis and total pleurectomy in treatment of patients with primary pneumothorax--early results]. AB - A group of 235 patients were treated between 1981 and 1993 for primary spontaneous pneumothorax. Immediate recovery was obtained in 150 (75%) out of 8 in whom drainage had been applied, and in a total of 79 who had been operated. Fifty of the remaining 52 patients with the first episode of pneumothorax, had repeated paracentesis with no complete lung expansion prior to operation, and 2 were referred to pleural cavity drainage. Three out of 79 operated patients, required re-thoracotomy (2 on the second day, and 1 on the eighth day following operation) because of hemothorax. In 3 patients an infection developed, thereupon a prolonged drainage of pleural cavity was used with good result. PMID- 9857645 TI - [Repeated paracentesis and total parietal pleurectomy in treatment of patients with primary pneumothorax--late results]. AB - Out of 235 patients discharged from the Department of Thoracic Surgery, Silesian Medical Academy, after treatment of primary spontaneous pneumothorax between 1981 and 1993, 232 (98.7%) answered our questionnaire and so were included in the study. Mean follow-up time was 10.4 years. No recurrence of the disease was reported by any of 79 patients who had been treated by surgical procedure/thoracotomy plus pleurectomy/, but 21 (14.3%) out of 147 treated by paracentesis reported a recurrence of pneumothorax. In 17 patients the recurrence was the first-time case, in 3 the second-time, in 1 the third-time. Out of 232 patients, 10 (4.3%) had a recurrence of pneumothorax on the opposite side. Achieving expansion of the lung, 80% of the patients reported no problems at all. Only 6 (2.6%) of the whole group stated they could no longer work in their profession because of the pneumothorax they had suffered in the past. PMID- 9857646 TI - [Secondary prophylaxis of venous thromboembolism: analysis of 49 fatal cases]. AB - The analysis of 49 fatal cases of venous thromboembolism--VTE (15% of total ambulatory patients number during long observation was performed. The advanced age of patients, multiple risk factors, underlying circulatory and respiratory tract diseases, malignancies, previous episodes of VTE especially with secondary pulmonary hypertension were the most important factors determining fatal prognoses in those patients. PMID- 9857647 TI - [Secondary ambulatory prophylaxis of venous thromboembolism: outcome of patients who were lost to follow-up]. AB - We have attempted to determine the outcome of 87 out-patients who were lost from follow-up. Several factors have been assessed: causes of lost from follow-up duration of oral anticoagulation, recurrent venous thromboembolic events, cause of death (if applicable). PMID- 9857648 TI - [The effect of chemotherapy on bone metabolism in patients with non-small cell lung cancer]. AB - In this study the influence of chemotherapy on bone metabolism in patients with III stage of non-small cell lung cancer has been evaluated. 30 patients (men) in average age 57.1 years were enrolled to this trial. The trial carried out on 2 groups of patients: group A consisted of 30 patients with NSCLC, group B of 15 patients without neoplastic process. In all participants bone scintigraphy was done and the following biochemical markers were measured: osteocalcin concentration, alkaline phosphatase's bone fraction, twenty-four-hour calcium and hydroxyproline urine excretion. In patients after two courses of chemotherapy (mitomycin, cisplatin, vinblastine) statistically significant reduction of serum concentration of osteocalcin, alkaline phosphatase, also reduction of calcium and hydroxyproline urine excretion have been observed. Scintigraphic examination of bones showed statistically significant decrease in resorption of isotope (MDP 99mTc) in patients after chemotherapy (p < 0.05). CONCLUSIONS: The study shows positive influence of chemotherapy on bone metabolism in patients with NSCLC. Patients treated with chemotherapy have bone metastases less frequently. PMID- 9857649 TI - [Evaluation of the prognostic significance of P53 mutation in patients with non small cell lung cancer]. AB - Prognostic value of p53 gene mutation was determined in 95 radically operated non small cell lung cancer patients (78 males and 17 females, mean age 57.8 years). Study group included 62 cases of squamous cell carcinoma, 30--adenocarcinoma and 3--large cell carcinoma. There were 52 patients in stage I disease, 16--in stage II, 26--in stage IIIa and one--in stage IIIb. Paraffin-embedded samples of resected tumors were assayed for p53 mutations with the use of PCR/SSCP analysis. p53 mutation were present in 22 cases (23%). The median survival in patients with and without p53 mutations were 49 and 75 months (p = 0.46), respectively, and the five-year survival rate 53% and 50%, respectively. In stage I disease the median survival for patients with p53 mutation was 53 months and for those without mutations the median survival could not be determined as more then a half of them were alive. Median survival in stage II patients with and without mutations was 35 months and 44 months (p = 0.62), and in stage IIIA--9.5 months and 17 months, respectively (p = 0.37). The results of this study indicate that p53 gene mutation is not correlated with prognosis in non-small cell lung cancer patients. PMID- 9857650 TI - [Typical and atypical pulmonary carcinoids--pathologic and clinical analysis of 77 cases]. AB - Neuroendocrine tumors of the lung represent a group of controversial pulmonary neoplasms regarding their classification, criteria for diagnosis, predictors of future behavior, and therapy. The histologic criteria for their classification rest heavily on the proposed by Arrigoni et al. in 1972, for atypical carcinoid (AC). According to these authors AC has mitoses between 5 and 10 per 10 high power fields (HPF), necrosis, hypercellularity and disorganized architecture. The survival analysis performed by Flieder et al. (1997) for a variety of clinical and histologic features revised the histologic criteria for separating AC from typical carcinoid (TC) and proposed a range of mitotic counts between 2 and 20 per HPF for AC. From 1978 until 1997, 77 resected pulmonary carcinoids were reclassified for TCs and ACs according to Flieder's et al. histologic criteria. The clinical and pathological various features were studied for the group of 62 TCs and 15 ACs. 77 patients (pts) entered the study: 29(38%) males and 48(62%)females; mean age 43 years, range 18-75 years, 46 pts underwent lobectomies, 16 bilobectomies, 12 pneumonectomies and 3 wedge resections. The patients with TC were younger than those with AC (males: 40 vs 50 years and females 42 vs 49 years). TCs were significantly smaller than ACs (mean diameter respectively: 24 mm vs 33 mm). Fifty four (87%) of TCs and all ACs had central localization. The time of patients observation ranged from 2 months to 18 and half years; 2 patients with TC died due to tumours progression; 3 due to other diseases. Regional lymph node metastases occurred in 10% TCs and 33% ACs (p = 0.032). The heterotopic bone formation appeared in 11(18%) TCs and 2(13%) ACs. The mitotic counts for AC range between 2 and 6 per 10 HPF, accompanied by small foci of necrosis in 2 cases. Peripheral carcinoids showed a spindle-cell morphology. The performed study has highlighted the new concept of the carcinoids classification and the importance of the mitotic counts as histologic criteria for AC diagnosis. The data based on the largest in Polish literature lung carcinoids collection. PMID- 9857651 TI - [Early and late results of surgical treatment in patients with primary mediastinal germ cell tumors]. AB - A group of 37 patients--20 (54%) men and 17 (46%) women--aged 2-65, mean 31.6 years, was treated between 1978 and 1995 for different kinds of primary mediastinal germ cell tumours. In 14 (37.8%) of them a mature solid teratoma was diagnosed, in 13 (35.1%)--dermoid cyst, in 5 (13.5%)--malignant teratoma, in 1 (2.7%)--immature teratoma, in 4 (10.8%)--seminoma. Mediastinal tumour was removed completely in 30 (81.1%) patients, partially in 7 (18.9%). Out of 37 patients, 3 (8.1%) died during postoperative time. Out of 26 patients discharged after surgical removal of mature teratomas, all are alive and have been followed-up for 10 years (17 patients) or 5 years (6 patients). Out of 8 discharged after surgical removal of malignant neoplasm, 3 are alive and all of them had primary mediastinal seminoma. One of them has survived 17 years after complete resection, and two patients--7.5 years and 2 years after partial resection and radiotherapy. Mean survival time is 4 years and 2 months for all patients with malignant neoplasm. PMID- 9857652 TI - [Procollagen III in bronchoalveolar lavage fluid of patients with allergic alveolitis and sarcoidosis]. AB - In 12 patients with sarcoidosis and 11 patients with allergic alveolitis concentration of procollagen III peptide in bronchoalveolar lavage fluid (in bronchial and alveolar fraction) was estimated using RIA method. In studied populations procollagen III levels were higher in comparison to control. In patients with allergic alveolitis and with DLCO < 60% pred. procollagen III peptide concentrations in BAL fluid were significantly higher than in patients with sarcoidosis. In patients with allergic alveolitis a positive correlation between BAL lymphocytes number and procollagen III peptide concentration was observed. PMID- 9857653 TI - [Mast cell: it's significance in bronchoalveolar lavage fluid cytologic diagnosis of bronchial asthma and interstitial lung disease]. AB - The analysis of mast cells in bronchoalveolar lavage (BAL) is rarely performed in routine diagnostic protocol. The BAL mast cells are rather difficult to be identified due to their paucity and technical problems with fixation and staining methods. The present study was carried out in order to characterize quantitatively and morphologically of mast cells obtained in bronchoalveolar lavage from 38 patients with several lung disorders. The percentage of mast cells ranged from: 0.0-0.6% (aspirin intolerant asthmatics), 0.1-1.8% (aspirin tolerant asthmatics), 0.9-1.8% (sarcoidosis), 1.5-2.0% (fibrosis), 0.04-0.08% (normals). There were statistically significant differences in the mast cells count between tested disease groups. Lung mast cells exhibited heterogeneity of size, shape, intensity of staining and degree of degranulation. Bronchoalveolar lavage is a very interesting and suitable model for the study of human lung mast cells. These cells should be analysed because of their clinical importance in pathogenesis of several lung disorders. We recommend, Carnoy's solution as a fixative and 0.5% solution of Toluidine blue (pH-0.5 with prolongation of staining time to 5-7 days at 4 degrees C) as a dye for routine diagnostic method of BAL mast cells identification. PMID- 9857655 TI - [Pneumonia and panophthalmitis as first signs of pneumococcal endocarditis]. AB - A case of pneumonia associated with pneumococcal endocarditis of bicuspid aortic valve complicated by pneumococcal meningitis and panophthalmitis successfully treated by antibiotics and late valve replacement was described. PMID- 9857654 TI - [Primary biliary cirrhosis with sarcoid-like infiltrations in the lung]. AB - A women, 50 years old, has been observed for 4 years because of recurrent infiltrations in both lungs. Biopsy of those lesions revealed sarcoid-like granulomas. Tubercle baccilli were not present in those lesions nor in the sputum or bronchial washings. Anti-tuberculosis therapy was without effect. On admission to our hospital she was in good performance state. There was an infiltration in the base of the left lung. Hepatosplenomegaly was observed on USG examination. Aspartate aminotransferase was 49 UI/l, alanine aminotransferase 70 UI/l. Alkaline phosphatase was 167 UI/l and the titer of antimitochondrial antibodies was 1:2000. Primary biliary cirrhosis was suspected, but the patient refused liver biopsy. Prednisone in the dose of 60 mg per day was given to suppress the granuloma formation in the lungs. During this treatment there was a decrease in size of liver and spleen, lung lesion disappeared and the titer of antimitochondrial antibodies decreased to 1:40. After 3 months of treatment the dose of prednisone was reduced gradually. When she was receiving 15 mg of prednisone every other day the titer of antimitochondrial antibodies rose to 1:8000 and the activity of alkaline phosphatase to 448 UI/l. At this time she accepted liver biopsy. Primary biliary cirrhosis was diagnosed. The possible connection between sarcoid-like granulomas in the lungs and the primary biliary cirrhosis is discussed. PMID- 9857657 TI - [Analysis of deaths during the course of status asthmaticus in patients treated at the Acute Asthma Center in the Pneumonology Clinic of the Warsaw Medical School in the years 1991-1996]. AB - In the Acute Asthma Treatment Center (OLSA) in the Department of Pneumonology of Warsaw Medical School in years 1991-1996, 582 patients with status asthmaticus were treated. The causes of status asthmaticus were bronchial asthma in 317 patients and COPD in 265 patients. Status asthmaticus was the cause of death in 21 patients treated in OLSA, which accounts for 3.6% of the total. 10 subjects were admitted with symptoms of brain death, who underwent resuscitation on their way to hospital. This study retrospectively analyzes the clinical characteristics (age, sex, PaCO2, pH, PaO2, time of mechanical ventilation and duration of treatment in ICU) of patients who died in status asthmaticus. They were divided into two groups: patients with asthma and COPD. A significant difference (p < 0.01) was detected between those two groups only in patients age. Mean duration of mechanical ventilation was 151 h in asthmatic and 104 h in COPD group. Mean duration of the OLSA stay was 19.5 days in the first and 7.4 days in the second group. The following fatal complications were observed: 2 cerebral strokes, 4 cardiac infarctions, 3 pneumothoraces, 2 atelectasis, 4 pneumonia, 1 case of gastric hemorrhage and 1 hemorrhage to mediastinum. PMID- 9857656 TI - [Rhodococcus equi--new human threat]. PMID- 9857658 TI - [The level of RANTES and interleukin-8 in serum of children with bronchial asthma after an exercise test]. AB - The study was carried out on 20 children with bronchial asthma. After the exercise induced test we observed decreased serum level of RANTES both in children with positive and negative results of the test. Serum level of IL-8 was increased in all examined group. We found strong correlation between serum level of RANTES and peripheral blood eosinophil counts in children with positive result of the test before and after the exercise. PMID- 9857659 TI - [Evaluation of the incidence of clinical symptoms and risk factors for COPD based on the monitoring of refinery workers in Piock (1993-1996)]. AB - Work place environment plays an important role in development diseases because of the time spend in and different toxic factors placed in it. The aim of study was to assess the ventilatory efficiency in individuals employed in Generating Plant (GP) in relation to the pollution of the work place and the tobacco smoking habit during 3 years. The study covered a group of 144 male individuals employed at GP. (age 39.3 +/- 8.7 yrs, period of employment 20.3 +/- 8.6 yrs, smokers 53.5%. Lung function tests consisted of VC, FVC, FEV1, FEV1%VC, PEF, FEF50 counted from flow volume curve and TGV, Rt measured using pletysmographic method. All tests were performed using pletysmograph (Masterlab) "Jaeger" placed on the ambulance near to the work place. The measurements were taken in May 1993 and 1996. Results were compared to with normal values (acc. ECSC). The anamnesis was obtained from all workers in the form of questionnaire projected for this study. All measurements were done during work time (9.00 a.m. to 2.00 p.m.). Mean values of the ventilation indices remained within the normal range but comparing results in smokers and non-smokers group significant differences in FEV1 and TGV were found. The symptoms of chronic bronchitis were present in 20.8% of persons. Spirometric criteria for the COPD diagnosis were found in 16 persons (11%). Only 5 persons (31%) were symptomatic. The mean decrease of FEV1 was 16.2 ml/yr, in the COPD group it was 82.5 ml/yr. The greater annual loss of FEV1 was found in the smokers and symptoms groups. PMID- 9857660 TI - [Gas exchange, blood acid-base balance and mechanical muscle efficiency during incremental levels of exertion in young healthy individuals]. AB - In this study we have evaluated the changes in gas exchange variables, blood acid base balance and the mechanical efficiency of muscle in healthy young men during an incremental exercise test. Twenty-six healthy men: age 22.1 +/- 1.4 (mean +/- SD) years, body mass 73.6 +/- 7.4 kg, height 179 +/- 8 cm, were subjects in this study. The subjects performed an incremental exercise test on a cycloergometer at a pedalling rate of 70 rev.min-1. The exercise test started at a power output of 30 W, followed by an increase of power output by 30 W every 3 minutes. Gas exchange variables were measured continuously (breath by breath). Antecubital blood samples for acid-base balance variables and plasma lactate concentration [La]pl were taken at the end of each 3-minute step. The lactate threshold (LT) in this study was defined as the highest power output above which [La]pl showed a sustained increase of > 0.5 mmol.l-1.step-1. The power output at LT amounted to 127 +/- 28 W. It corresponded to 45% of the maximal power output (MPO) reached at maximal oxygen uptake (VO2 max). The oxygen uptake at the LT amounted to 1734 +/- 282 ml.min-1 and corresponded to 48% of VO2 max (3726 +/- 363 ml.min-1). The minute ventilation at the LT amounted to 47.8 +/- 7.5 l, and its increase to the level of 125.7 +/- 19.7 l reached at the MPO was obtained mainly by intensification of breathing frequency from 23.8 +/- 3.31.min-1 to 43 +/- 5.91.min-1, for LT and MPO respectively. Analysis of the changes in PETCO2 during the incremental exercise test showed significant differences between subjects. One could recognise a group of subjects (n = 8) with high values of PETCO2 (above 45 mmHg) and a group of subjects (n = 8) with lower values of PETCO2 (below 43 mmHg). However, no significant differences in exercise tolerance, expressed by the level of MPO and maximal oxygen uptake, were found between those groups of subjects. The mechanical efficiency calculated on the basis of power output/net oxygen uptake ratio during cycling at a power output of 60 W amounted to 24.1 +/- 3.8 percent, at the LT 25.8 +/- 2.1%, whereas at the maximal power output a significant (p < 0.01) drop in muscle efficiency occurred, to the value of 23.1 +/- 1.6%. This drop in muscle efficiency occurring at the MPO may be an important factor limiting exercise tolerance when performing high power output exercise. IN CONCLUSION: The above presented data illustrate the physiological responses to incremental exercise and the level of exercise tolerance, which may serve as a reference point for the population of healthy, young physically active Polish students. PMID- 9857661 TI - [Changes in platelet function, activated partial thromboplastin time and plasminogen activator inhibitor in patients with bronchial asthma after prednisone treatment]. AB - Twenty patients with recently recognised bronchial asthma and 22 age and sex matched healthy control subjects were studied. In both groups the activated partial thromboplastin time (APTT), prothrombin ratio (INR), fibrinogen, euglobulin lysis time (ELT), platelet number and platelet adhesion and aggregation, tissue plasminogen activator antigen (t-PA Ag) and activity of plasminogen activator inhibitor (PAI-1) were tested and compared. Statistically significant differences between asthmatic and control groups concerning adhesion, aggregation, APTT and ELT were found. In asthmatic group after 14 days of prednisone administration in a dose of 20 mg/d statistically significant (p < 0.05) shortening of APTT, and a significant increase of adhesion, aggregation and PAI-1 activity were found. These results suggest that in asthmatic patients after prednisone treatment platelet activity appeared in a form of intensification of adhesion and of aggregation degree, also the activity of PAI-1 probably of the platelet origin increased. PMID- 9857662 TI - [Nitric oxide in expired air from rats with experimental bronchitis]. AB - Bronchitis was evoked in 20 rats by SO2 inhalation. Ten rats were exposed to SO2 only, 10 others were exposed to SO2 and treated with heparin simultaneously. The control group consisted of other 10 rats. NO concentration in the air exhaled by all animals was determined. The experiment was carried out in a specially constructed chamber. It lasted 15 weeks. In the group of animals exposed to SO2 only, NO concentration was found beginning from the %th week, with the maximum concentration in the 10th week. In the group of animals exposed to SO2 and treated with heparin NO concentration was much lower than in the above group. NO was not found in the control group. The conclusion is that NO concentration in the exhaled air is a result of bronchial inflammation, and can be decreased during treatment with heparin. PMID- 9857663 TI - [Urokinase plasminogen activator /U-PA/in blood of patients with lung cancer]. AB - Clinical and experimental studies suggest that urokinase plasminogen activator/u PA/ is associated with cancer invasion and metastasis. The aim of study was to evaluate the concentration and activity of u-PA in the plasma of patients with planoepithelial lung carcinoma. The examined group consisted of 40 patients/33 men, 7 women/aged 58 years with lung cancer in II, IIIa and IIIb stage of disease and 30 healthy adults as control. In citrate plasma concentration and activity of u-PA with ELISA method were performed. Concentration of u-PA was significantly lower in cancer patients then in controls. No associations between sex, age and plasma levels of u-PA were observed. Concentration and activity of u-PA in patients with IIIb stage of cancer were significantly decreased in comparison to patients with IIIa stage. PMID- 9857664 TI - [Treatment with salmeterol and quality of life in patients with asthma]. AB - The aim of this study was to assess an influence of 4-week treatment with salmeterol on quality of life in asthmatic patients. It was a double blind, placebo controlled study. On the first visit spirometer parameters and PC20H were measured. Within one week of observation all patients recorded scores of asthma symptoms and after that time 22 patients were randomised to the treatment with salmeterol or placebo. On the fourth visit 4-week salmeterol or placebo therapy was finished and all parameters were carried out as on the first visit. Symptoms severity recorded in diary cards was reduced after 4-week of salmeterol therapy. Four week treatment with salmeterol did not change PC20H. Treatment with salmeterol reduced severity of asthma symptoms and use of short-acting beta 2 agonists. The reduction of asthma severity and the use of bronchodilators was more pronounced in the group of patients treated with salmeterol than in the placebo group, but the differences were not significant. PMID- 9857665 TI - [Results of examination in healthy volunteers of rifampicin and isoniazid bioavailability from Polish two-drug combination capsules of rifamazid used for tuberculosis treatment]. AB - Purpose of the study was the examination of the relative bioavailability of rifampicin and isoniazid from Rifamazid (fixed-dose combination) and bioavailability of each drug in free simultaneous combination. The study was carried out as an open randomized cross-over study comprising 16 healthy volunteers for study of rifampicin and 15 healthy volunteers (slow acetylators) for isoniazid. The results have shown, that the pattern of absorption, elimination, plasma concentrations and pharmacokinetics parameters were found similar after administration of the drugs in free and in fixed combination. PMID- 9857666 TI - [Pneumocystis carinii infections in patients with neoplasms and AIDS--clinical pathologic changes in the lungs]. AB - Pneumocystis carinii (PC) has been documented as a cause of pneumonia in a broad range of immunocompromised patients. The presentation of disease will very based on the underlying predisposing condition but a confirmation depends upon the identification of organisms in a bronchial aspiration or lung biopsy specimen. This retrospective study based on autopsies of 15 patients (pts) with AIDS and 12 non AIDS pts with neoplastic diseases. Pneumocystis carinii pneumonia (PCP) was confirmed by histologic examination. The clinico-pathological analysis emphasizes a spectrum of morphologic variation of the disease in relation to the clinical course of a principal disease. A distinction was made between the microscopical diagnosis of PC infection in AIDS and non AIDS pts; the burden of organisms in infected AIDS pts appeared greater than that of neoplastic diseases (mostly with small cell lung carcinoma). Nonspecific features of diffuse alveolar damage associated with PC organisms were identified in 67% of non AIDS pts and 13% of AIDS pts. Various degree of interstitial fibrosis was more prominent in AIDS pts (67%) than in non AIDS pts. The high frequency of atypical changes in lung might be the result of various chemotherapeutic agents used in managing these pts or repeated pulmonary infections. PMID- 9857667 TI - [Can an aortic anomaly cause intense dyspnea in a patient with bronchial asthma?]. AB - 64 old man with bronchial asthma and a double arch of aorta was presented. Clinical symptoms and value of flow-volume curve before and after beta 2 agonist inhalation for differential diagnosis in this case were discussed. PMID- 9857668 TI - [Invasive pulmonary aspergillosis in the course of allergic pulmonary alveolitis]. AB - 20 years old man with history of acute allergic alveolitis 7 years ago, was admitted to hospital because of dyspnea, fever and cough after massive exposition to organic dust. Corticosteroids were introduced. During 3-weeks treatment respiratory failure progressed. Chest x-ray showed massive bilateral nodular opacities. Antibiotic therapy was not effective. The autopsy revealed invasive pulmonary aspergillosis. Presence of precipitins to aspergillus fumigatus was confirmed postmortem. Diagnostic difficulties are discussed. PMID- 9857669 TI - [Endometriosis of the lung diagnosed with videothoracoscopy]. AB - A case of non symptomatic pulmonary endometriosis appearing as multiple nodules on chest X ray and chest CT scan is reported. 43 year old woman was admitted to surgical department because of suspicion of neoplastic dissemination of unknown origin. Non invasive diagnostic procedures were not effective. Diagnosis of pulmonary endometriosis was established with thoracoscopy with lung sampling, confirmed with histopathologic evidence. PMID- 9857670 TI - [Pathophysiologic programming of cell death]. AB - In multicellular organisms homeostasis depends on a balance between cell proliferation and cell death. In this review principles of the physiology of programmed cell death (apoptosis), i.e. biochemical features, involved genes and proteolytic enzymes, are described. Alterations in apoptosis contribute to the pathogenesis of a number of human diseases, including cancer, viral infections, inflammation, hematopoietic and immunological system defects (e.g. AIDS), neurodegenerative disorders. Specific effect on regulation of apoptosis might lead to new possibilities for treatment. Methods of quantitative determinations of apoptosis are discussed. PMID- 9857671 TI - [Animal lectins--structure and function]. AB - Lectins are structurally and functionally diverse group of proteins or protein domains capable of specific binding of oligosaccharide structures present on cell surfaces, the extracellular matrix, and secreted glycoproteins. Animals lectins are classified into six groups: C-type lectins, S-type lectins (galectins), I type lectins, P-type lectins, pentraxins, and others. In this review, the basic knowledge regarding structure and function of animal lectins is presented. PMID- 9857672 TI - [The role of thrombin in hemostasis]. AB - Thrombin is a serine protease that plays a central role in hemostatic and thrombotic events. The structure, different forms of thrombin, its role in platelet activation and in other cellular effects are reviewed. Two distinct receptors involved in platelet activation induced by alpha-thrombin are presented: a high affinity (GPIb) coupled to phospholipase A2 and a moderate affinity, G protein associated with seven transmembrane domain receptor (STDR) coupled to phospholipase C. PMID- 9857673 TI - [Biological properties and clinical significance of interleukins 12 (IL-12)]. AB - IL-12 is a heterodimeric cytokine composed of 2 disulfide-linked subunits with molecular masses of 40 and 35 kDa, respectively. The cytokine is produced by phagocytic cells, professional antigen-presenting cells such as dendritic cells, skin Langerhans cells and B cells. IL-12 production is induced by bacteria, intracellular pathogens, fungi, viruses, or their products in a T-cell independent pathway or a T-cell-dependent pathway, the latter mediated through CD40 ligand-CD40 interaction. Interleukin 12 induces interferon gamma secretion by T cells and natural killer cells, enhances the proliferation of activated T cells and natural killer cells, augments the cytolytic activity of cytotoxic T lymphocytes and natural killer cells, and supports the differentiation of Th1 helper effector cells. The therapeutic potential of these activities is suggested by studies in tumor and microbial models. IL-12 has suppressed tumor growth in all murine models examined. Antimicrobial activity has been demonstrated in bacterial, parasitic, and viral models of infection. The cytokine also stimulates in vitro antitumor activity of lymphocytes from patients with cancer. Current data indicate that CD4 T cells, CD8 T cells, natural killer cells and interferon gamma may contribute to the antitumor effects of interleukin-12 therapy. Clinical trials are being initiated to evaluate the possible therapeutic uses of IL-12 in the treatment of neoplastic diseases and some infections. PMID- 9857674 TI - [The significance of beta 2-microglobulin in diagnosis and therapy]. AB - This article describes the significance of beta 2-microglobulin (beta 2m) determination both in urine and serum for the estimation of proximal renal tubular function and glomerular filtration rate. The usefulness of beta 2m in therapy, cancer diagnosing, epidemiologic investigations of industrial exposure to cadmium, lead, mercury and in evaluating etiologic factors of environmental nephropathies was also discussed. PMID- 9857675 TI - [The role of cystatin in neoplastic progression and diagnosis]. AB - Possible role of the cystatins, natural and specific inhibitors of the cysteine peptidases in cancer invasion is described. The value of cystatins determinations in serum and tissue extracts as diagnostic and prognostic parameters in human cancers is also presented. PMID- 9857676 TI - [Mice with mottled mutation--a model for defective copper metabolism in humans]. AB - The group of X-linked mottled (Atp7aMo) mutations in mice is described. A normal gene encodes a copper-binding P-type ATPase. Mutant animals have the disturbance in copper metabolism, hemizygous males (Mo/y) die between 14-18 days of life, heterozygous females (Mo/+) are normal and fertile. This kind of copper metabolic defect is observed also in other animal and in human. In human Menkes disease caused by X-linked Atp7a mutant gene leads to death in early childhood. Because of is 89% of homology between Atp7aMo gene and Atp7a locus in human, mottled mutations are an excellent model for Menkes disease. PMID- 9857677 TI - [Infantile colic as maternal reason of consulting a pediatrician: characteristics of mother and child]. AB - About 10-20% of infants under four months of age suffer from infantile colic (IC), an excessive screaming behaviour caused by multiple factors. WESSELS et al. (1954) definition is that babies in usually good health and nutritional condition are screaming/whining more than three hours daily, more than three days weekly for no apparent reasons. The question is whether/in what way daily routines of infants concerned differ from normal infants. Mothers of colicky infants (n = 25) and mothers in a control group (n = 25) record characteristics of the mother/child relationship, their assumptions why the child is screaming, care and attention they provide and babies' reactions. For seven days the mothers kept a diary about the babies' behaviour and worked on questionnaires concerning social support received, depressant effects and exhaustion. Interviews about the extent of care provided as well as personal thoughts and emotions were conducted. The "colic group" formed two subgroups: one fulfilling Wessels criteria (Wessel Koliker; WK), the other more conforming with the "control group" babies though mothers complained about screaming (Non-Wessel-Koliker; NWK). "Colic group" babies received more attention and care (compared to the "control group" babies), attempts to calm them down frequently failed and mothers in this group reacted less promptly to their babies' screaming. "NWK-group"-mothers strongly tend to attribute the babies' screaming to "meteorism" or "discontent". No differences existed between groups concerning social support, depressant effects and exhaustion. There is a risk that the colic may negatively influence the mother/child interaction beyond three months. PMID- 9857678 TI - [Computer assisted training of attention and concentration with healthy children]. AB - The intention of this study was to analyze in how far a computer assisted training for brain damaged adults can be used for the practice with children. Healthy children of different ages were trained with this attention and concentration programme. The results should help to adapt the training programme for the use in brain damaged children. Age-groups differed significantly in level of performance. Data showed that in comparison to Kindergarten-children, first grade children showed a higher benefit from the training. Implications for modifications of the training-programme are discussed. Other parameter of performance (reaction-time, errors) in relation to school age provide clues to the distribution of structure of difficulty in the programmes. A questionnaire assessing motivation before and during the training, intelligibility of the task, feasibility of the computer and adequacy of the test in children showed that sustained motivation can be achieved provided the training is used in an adaptive way. Thereby demotivating with too easy tasks as well as frustration caused by difficult tasks can be avoided. In general, there was good acceptance of the computerised cognitive training. The presence of a therapist during every training session proved to be necessary in children under the age of 6 years. PMID- 9857679 TI - [Evaluation of a group therapy program for anxious children]. AB - In this study 14 children with different anxieties attended in small groups a cognitive-behavioral training program. The groups met once a week over a period of ten weeks. Outcome was evaluated with different questionnares for children and parents and interviews at the beginning and the end of training and ten weeks later. The different measures showed a significant reduction of self-reported anxiety. Anxious, avoident and social insecure behaviour was also reduced. Further the negative self-talk of the children changed into more appropriate positive self-talk. As it was not possible to use an appropriate control group, the results can just be seen as hints for the effectivity of a cognitive behavioral orientated group therapy program for the treatment of various childhood anxieties. Further research under controlled conditions is necessary. PMID- 9857680 TI - [Child and adolescent psychiatric services in the canton of Zurich: a comparison of outpatients, day clinic, and inpatient referral populations]. AB - Findings in intervention research are an important basis for the discussion of issues pertaining to quality control. In Switzerland such findings have been rare. The present study of the entire 1988-1995 referred child and adolescent population of the Child and Adolescent Psychiatric Service of the Canton of Zurich (N = 7538 treated patients) concentrated on the demographic, diagnostic, and therapeutic attributes of the outpatient, day clinic, and inpatient populations that comprised the referred population. Marked differences are found in comparable studies from Germany with regard to patients treated in an inpatient setting. For example, the mean age of the patients in the Zurich sample is lower and patients with conduct disorders are more frequently represented. On the other hand, patients with psychotic illness are missing and, on the average, the duration of treatment is considerably longer. The reasons for these differences are to be found in the various structural attributes of the cantonal services that are provided in Zurich. In the area of inpatient care, where in comparison to outpatient settings and also day care settings patients with severe disorders are treated, there is a lack of places for the short-term care of acutely ill patients--for example, psychotic patients. This is especially true for adolescent patients. The limitations of traditional outpatient settings are frequently marked by unsatisfactory treatment results in outpatients with conduct disorders, developmental disorders, and a high impact of abnormal psychosocial circumstances. PMID- 9857681 TI - [No therapy possible? A group therapy model for psychologically disturbed and socially disadvantaged children]. AB - The authors have developed a group therapy model specifically for children who are difficult to treat using traditional psychotherapeutic techniques. In the present article they describe this model, which has been employed successfully for the past four years in an elementary school in an area of high social deprivation, in collaboration with teaching staff and a social worker. PMID- 9857682 TI - [Nephrology in the first decade of the twenty-first century]. AB - Although prophecy in medicine is extremely difficult, I do not expect, that xenotransplantation will become reality in the first decade of the 21th century. However I expect introduction of safer and more effective immunosuppressive agents or even induction of selective immunologic tolerance in kidney transplant patients. I expect a significant decline in the number of uraemic patients with diabetic or hypertensive nephropathies, thanks to better prevention and treatment of these pathological states. I predict intensification of studies of renal fibroblasts responsible for progression and loss of renal parenchyma in nephropathies of different etiology. Gene therapy will become a reality both in inborn and acquired nephropathies although safety of this kind of therapy remains to be established. Finally I expect essential improvement of therapy in clinical settings induced by abnormal homeostatic function of the kidneys mostly thanks to our better understanding of the molecular background of this function. PMID- 9857683 TI - Immunopathogenesis of glomerulonephritis; the importance of new concepts for clinical practice. PMID- 9857684 TI - The immunopathogenesis of IgA-nephropathy. PMID- 9857685 TI - Biology of cultured podocytes. PMID- 9857686 TI - The role of blood platelets in glomerulonephritis. PMID- 9857687 TI - Progression of renal damage in chronic glomerulonephritis and the therapeutical implications. PMID- 9857688 TI - Advanced glycation endproducts and the kidney. PMID- 9857689 TI - Dialysis therapy in the region of central and eastern Europe. PMID- 9857690 TI - Hemodialysis and CAPD--are they comparable? AB - I have made comparisons of thrice weekly HD (not daily HD) and PD (mainly CAPD). In terms of patient survivals, there is no proven modality advantage. In terms of cost and quality of life, PD probably has advantages in many patients. With a tendency towards earlier initiation of dialysis and incremental prescriptions to achieve a targeted amount of small-solute clearances, PD probably has cost and technical advantages. PMID- 9857691 TI - Contemporary points of view on treatment of acute renal failure (ARF). PMID- 9857692 TI - Non-immune mechanisms of progressive graft dysfunction. PMID- 9857693 TI - Relationships between dialysis adequacy, dietary intake and nutritional status of patients treated with continuous ambulatory peritoneal dialysis (CAPD). PMID- 9857694 TI - Urinary tract infections--pathogenetical aspects. PMID- 9857695 TI - Selected problems concerning the diagnosis of urinary tract infection. PMID- 9857696 TI - Chemotherapy of urinary tract infections. PMID- 9857697 TI - Urinary tract infections and renal stone disease. PMID- 9857698 TI - Urinary tract infections in children. PMID- 9857699 TI - [Can we prevent late complications of urinary tract infections?]. AB - Urinary tract infections are one of the most common renal diseases sometimes leading to renal injury and in consequence to chronic renal failure. The most frequent causative pathogen responsible for this infection is Escherichia coli. There are several factors which increase the risk of infection including vesicoureteral reflux, cystic renal disease, urinary calculi, obstruction and other anatomical and functional abnormalities of urinary tract as well as neurological bladder dysfunction, long term indwelling catheters, mechanical vaginal diaphragms and intensive sexual intercourse. This paper will highlight general view on the treatment of different manifestations of urinary tract infections including asymptomatic bacteriuria, urethritis, cystitis, prostatitis as well as acute and chronic pyelonephritis. The details of those problems will be elucidated in another paper. PMID- 9857700 TI - Current trends in the treatment of hypertension in renal disease. PMID- 9857701 TI - Hypotensive therapy in diabetic renal disease. PMID- 9857702 TI - Compliance of patients with arterial hypertension to therapeutic recommendations. PMID- 9857703 TI - [Treatment with erythropoietin in Central and Eastern Europe]. AB - Human recombinant erythropoietin (r-HuEpo) was introduced widely to the clinical practice more than ten years ago. In many countries belonging to Central and Eastern Europe (CEE) use of this drug was very delayed mainly due to the high costs of treatment. The aim of the presented study was to estimate actual possibilities of r-HuEpo administration in thirteen CEE countries. Survey was performed using especially prepared questionnaires including questions connected with r-HuEpo and filled out by the members of the Central and Eastern Advisory Board in Chronic Renal Failure. Below mentioned numbers of patients were under the control of the responders during performing the survey: on hemodialysis (HD) 1119, on peritoneal dialysis (PD) 205 and 1556 after renal transplantation (RT). Respectively in subsequent groups 53.5% (HD), 41.5% (PD) and 16.2 (RT) of patients were treated using r-HuEpo. Possibilities of treatment differs significantly between two groups of countries. In those with better developed economy (Croatia, Czech, Hungary, Macedonia, Poland, Slovakia, Slovenia, Yugoslavia) 60-70% of dialysed patients were given r-HuEpo and in the rest of countries (Bulgaria, Latvia, Lithuania, Romania, Russia) this drug was administered in 10-35% of patients. The most popular way of the r-HuEpo administration were subcutaneous injections (89.3%) comparing with only 10.7% of patients getting this important hormone intravenously. Average weekly dose of r HuEpo was 71-90 IU/kg/week in so treated patients and 91-100 IU/kg/week in whom drug was administered i.v., in majority of patients two or three times weekly. Major part of responders tried to achieve as the optimal level of Hb 10-12 g/dl but simultaneously in 1/3 of centres only achieving of the Hb level 9-10 g/dl was possible. The general majority of responders were not satisfied with the obtained results and recognised economical constrains as the main reason of such a situation. These problems were more widely observed in predialysis patients, because only few of them were on r-HuEpo treatment. Side effects of this drug expressed rarely and main of them were development or acceleration of hypertension (16%) and thrombotic complications in the vascular access (8%). The main cause of hyporesponsiveness to drug was iron deficit despite of the adequate monitoring of responders were convinced that further avoiding of economical constrains would be benefit dialysis and predialysis patients. It is especially important looking on the literature finding supported by personal experience showing that r-HuEpo was positive impact on the quality of life, status of the cardiovascular system, endocrine, metabolic and immunological disorders observed in uremic patients. PMID- 9857704 TI - Iron therapy: overview and recommendations. PMID- 9857705 TI - [The usefulness of normalized protein catabolic rate (NPCR) and serum albumin concentration measurement for evaluation of clinical status in patients treated with continuous ambulatory peritoneal dialysis]. AB - Malnutrition is a serious and common problem among patients dialyzed with peritoneal dialysis. Simple and reproducible methods of nutritional status assessment are needed for early problem identification to prevent and treat this complication. The aim of the present study was to evaluate the usefulness of albumin serum concentration and normalized protein catabolic rate (NPCR) derived from urea kinetic modeling session to assess the clinical status of ESRD patients treated with Continuous Ambulatory Peritoneal Dialysis (CAPD). Statistically significant, negative correlations between NPCR and hospitalization rate (r = 0.307; p < 0.05) and co-morbidity score (r = -0.429; p < 0.001) as well as statistically higher NPCR in the group of patients who survived the whole study period (0.82 vs. 0.74 in Non-Survivors; p < 0.01) support the value of this marker as an indicator of clinical status of peritoneal dialysis patients. Significant, negative correlations between serum albumin level and co-morbidity score (r = -0.379; p < 0.05), peritonitis rate (r = -0.359; p < 0.05) and hospitalization rate (r = -0.601; p < 0.005) were also found. Low absolute values of NPCR for the whole population as well as significant, positive correlations of this marker with dialysis adequacy parameters with concomitant lack of such correlations for albumin indicate, that NPCR as a nutritional marker should be interpreted with caution, and always regarded to the calculation method. PMID- 9857706 TI - [Antibodies to glutamic acid decarboxylase (GADA) as markers of risk for development of insulin-dependent diabetes mellitus]. AB - Autoantibodies to glutamic acid decarboxylase (GADA)-found frequently before the onset of IDDM are an important marker of the autoimmune process of B cells destruction. The aim of the present study was to evaluate the possible relationship between the frequency of GADA and ICA in first degree relatives of subjects with IDDM. The GADA were determined in 238 first degree relatives of IDDM subjects. In 32 of the investigated subjects GADA were measured second time after 12 months and in 24 of them IVGTT was performed to evaluate FPIR and GADA prevalence relationship. The results demonstrate that combination of ICA and GADA antibodies has a high sensitivity and increases the specificity for predicting IDDM in high risk siblings. PMID- 9857707 TI - [The effect of vitamin E on antioxidant tissue activity in pregnant rats with streptozocin-induced diabetes]. AB - The aim of our study was to establish if moderate doses of nonenzymatic antioxidant, vitamin E, could play a role in the antioxidant defence system in pregnant diabetic rats and their offspring. Two models of experimental diabetes were used: the 1st group consisted of 30 female Wistar rats which were given a single dose of streptozotocin (40 mg/kg) and were mated 7 days later, whereas in the 2nd group diabetes was induced on the 7th day of pregnancy (using the same dose of streptozotocin). In each part of the experiment diabetic animals were divided into two equal groups: vitamin E supplemented and fed with standard diet only. Our controls consisted of 20 normal rats. On the 1 day after delivery rats were decapitated and homogenates of maternal liver and uterus as well as neonatal lungs and liver were prepared. Then the following parameters were measured: malondialdehyde (MDA) concentrations in the homogenates and blood serum (colorimetric assay), glutathione (GSH) levels (colorimetric test), the activity of CuZn superoxide dismutase (SOD) (spectrophotometric method) and glutathione peroxidase (GPx) (immunoenzyme technique). Statistical analysis was performed using Mann-Whitney U test. The neonates of diabetic rats were smaller than healthy ones and serum glucose concentration was markedly higher in diabetic animals, especially in model I. MDA levels increased significantly, whereas GSH content and SOD as well as GPx activities were markedly diminished in diabetic pregnant rats and their offspring in comparison with the control group. In animals supplemented with tocopherol MDA concentrations declined significantly, GSH content and SOD activities were markedly elevated in almost all types of tissues studied, whereas glutathione peroxidase remained suppressed. CONCLUSIONS: By monitoring the activity of chosen scavenging enzymes in homogenates and body fluids we can accumulate information on ongoing biological oxidative stress. Our results suggest that diabetic pregnant rats and their neonates are exposed to an increased oxidative stress, but vitamin E supplementation could at least in part reduce its detrimental effects. PMID- 9857708 TI - [Organization of services for treatment of postoperative pain--3-year experience]. AB - We report organisation principles and three year experience of Acute Pain Service in general surgery clinic. 481 patients were treated after abdominal and vascular interventions, hemorrhoidal varices and mammectomies. Continuous epidural, combined spinal-epidural, intrapleural anaesthesia and continuous brachial plexus block were used for pain control. Time of analgesia varied from 1 to 4 days. The level of analgesia was assessed as good (VAS 3) in 94.8% of cases. Complications were mainly technical due to catheter or antibacterial filter failure. In 2% of cases cardiovascular complications were observed. Respiratory depression occurred in 1 patient. The work of APS team was assessed as very good by both surgeons and patients. PMID- 9857709 TI - [Management and diagnostics in blunt abdominal trauma in pregnancy]. AB - Vital activity of pregnant women as well as development of motorization promotes abdominal traumas. This paper describes mechanisms of the most frequent traumas, specificity of history taking and physical examination in pregnant woman after injury as well as general condition assessment. Usefulness of the most commonly performed biochemical tests, diagnostic abdominal lavage and imaging methods is discussed. Therapeutic management resulting from the patients condition assessed and diagnostic procedures performed is also presented. PMID- 9857710 TI - [Prevention using folic acid--a good method for reduction of neural tube defects in Poland]. AB - Incidence of neural tube defects in Poland in the 90's was 2.68 in 1000 births. The value of incidence has not been changed within last twenty years. Mortality caused by neural tube defects in Poland is much higher than in many other European countries and the United States. High rates of incidence of neural tube defects in Poland is probably caused by low utilisation of methods of secondary prevention in a group of low-risk pregnant women. Since methods of primary prevention of neural-tube defects are available; primary prevention with folic acid, should be immediately implemented in Poland. PMID- 9857711 TI - [Inherited hyperammonemia]. AB - Inherited hyperammonemia disorders are caused by specific enzymatic defects in the urea cycle or in metabolic pathways related to it. These disorders can be divided into the following groups: deficiencies of urea cycle enzymes, transport defects of dibasic amino acids, organic acidemias, defects in beta-oxidation of fatty acids, transient hyperammonemia of the newborn-probably a not genetically determined disorder. Manifestation of the mentioned disorders includes elevated serum ammonia level resulting in altered level of consciousness and/or persisted vomiting. Occurrence of irreversible neurologic sequelae depends mostly on the extent of hyperammonemic period. Differential diagnosis includes blood gas, anion gap, plasma amino acids analysis and urine organic acids analysis. In some cases specific tissue enzymes activity measurement is necessary. Dialysis, sodium benzoate, sodium phenylacetate and arginine are used in the treatment of acute hyperammonemia. In addition oral or rectal neomycin and/or lactulose can be used, which reduces intestinal ammonia production. PMID- 9857712 TI - [Gastric motor dysfunction in diabetic patients: pathogenesis, clinical manifestations, diagnosis and treatment]. AB - Recent reports provide evidence, that the disordered gastric motoric function in diabetic patients depends on glycaemia and that it can influence metabolic control. The etiopathogenesis of disordered gastric motoric function is poorly understood and is mainly hypothetical. The diagnosis of this dysfunction is difficult. It is recommended to evaluate of gastric motility, especially by scintigraphy, manometry and electrogastrography. In the treatment, it is important to remember about the good metabolic control of diabetes and use the gastrokinetic drugs in the combination or alternating therapy. In especially severe cases, surgical therapy is indicated. Lack of effective method of gastric motoric dysfunction treatment in diabetic patients suggest the necessity of further studies on new drugs, which would normalise the motoric abnormality and eliminate the symptoms. PMID- 9857713 TI - [Celiac disease in insulin-dependent diabetes mellitus]. AB - A coincidence of celiac disease and insulin-dependent diabetes mellitus has been described in several studies. Celiac disease appears to be more common among patients with IDDM than in general population. The diagnosis of celiac disease requires histologic evidence of villous atrophy and a positive antibody test result. Typical gastrointestinal complaints of celiac disease are rare in IDDM patients, while atypical isolated symptoms of celiac are more common. It is recommended that all young diabetic patients, even those asymptomatic, should be screened for celiac. Routine screening is recommended repeatedly during first years after the diagnosis of IDDM. PMID- 9857714 TI - [Peri-renal pheochromocytoma]. AB - Presented is a case of pheochromocytoma with an ectopic or extrarenal localisation in a 47 year old female farmer. The patient was admitted to the 1st Chair and Clinic of Internal Disease, Jagiellonian University Collegium Medicum due to a three year history of hypertension. The bouts of hypertension were characterised by sudden increases of blood pressure to 250/150 mmHg, which occurred after physical exertion but particularly during the night. On physical examination no abnormalities were detected. Laboratory analysis revealed no changes apart from mild hyperglycemia. 24 hour vanillymandelic acid excretion was normal on two successive accounts, but on a third analysis was raised. Fundus examination revealed bilateral hypertensive and diabetic angiopathy. Abdominal ultrasound revealed a tumor in the region of the pancreas with an echogenicity similar to that of the liver. Abdominal CT scan revealed normal suprarenal glands bilaterally. A smooth walled tumor of 55 mm diameter was visualized below the head of the pancreas. Its structure was not uniform with degenerating features. The tumor adhered to the descending and transverse part of duodenum, compressing the vena cava inferior and was in close proximity to the abdominal aorta. After surgical consultation the patient was transferred to the II Clinic of General Surgery Jagiellonian University Collegium Medicum where after pretreatment with alpha and beta blockers, ACE inhibitors, diuretics and Insulin CHOS the patient underwent surgery. Posterior to the duodenum, inferior to the head of the pancreas, between the aorta and vena cava inferior was a polycystic tumor of 80 mm diameter, which was evacuated in its entirety. Histopathology revealed a paraganglioma with an appearance of pheochromocytoma. On a three month follow up visit the patient was without symptoms. Her blood pressure and glycemia were normal despite being without treatment. PMID- 9857715 TI - [A giant gastric trichobezoar]. AB - The paper presents a 17-year-old girl with a big trichobezoar in the stomach, diagnostic and treatment proceeding. In our opinion upper GI tract endoscopy is the most useful adjunct to the diagnosis of all types of stomach besoars. PMID- 9857716 TI - [Public health matters in the early-Capitalistic period (XVII-XIX century]. AB - Modern opinion about people community formulated by philosophers and economists has shown essence of the public health care problem. Together with development of medical knowledge about man and his social role, sense of understanding joint responsibility of the government for health matters of the nation has been changing. Political and economical transformations in the era of early industrialisation tiggered in people need for making their material and health state better. This base was strenghtened by the French Revolution in proclaimed rights to freedom and individual life in the society. Modern adjusted to the epoch inner structures of undergoing modernization European countries, considered issues of the public health care in wider range. Sanitary, hygienic and epidemic problems as well as new health- and life-threatening factors in working population and pathology. PMID- 9857717 TI - [A little about the nineteenth-century history of a health resort in Ciechocinek]. AB - Impression of a Warsaw physician, hygienist and social worker Ludwik Natanson (1822-1896) from his journey to Ciechocinek in 1853 was reported. Brief history of this resort as well as contemporary indications and contraindications to treatment there were described. A role which in creation of this resort was played by Roman Ignatowski (1805-1889), who was a resort-doctor for many years, was presented. PMID- 9857718 TI - [Reductionism once again. The example of Helicobacter pylori]. AB - The contrasting evidence for a multifactorial pathogenesis of gastroduodenal disease rather than the reductionistic, monocausal role of Helicobacter pylori is presented. Evidence for the former is derived from epidemiological, physiological, immunological and experimental behavioral studies in animals. The high prevalence of the bacterium in popularies and the low incidence of peptic ulcer strongly suggests that it alone cannot play the only pathogenetic role. The evidence that peptic ulcer is not one disease raises the problem of identifying the contributions of psychosocial factors in combination with infection by H. pylori in the various forms of the human disease. The most likely role it plays is that of an "opportunist", when the gastroduodenal mucosal "defense" is compromised by many possible factors. PMID- 9857719 TI - [Psychotherapy research--how it should (not) be done. An expert reanalysis of comparative studies by Grawe et al. (1994)]. AB - 22 psychotherapy outcome studies used in the Bernese meta-analysis by Grawe, Donati and Bernauer (1994) for a treatment comparison between behavioural and psychoanalytic-psychodynamic treatment concepts were reanalysed by 12 expert psychotherapy researchers independent of each other. Three clinical criteria served as basic criteria for the assessment of the methodological quality of the 22 comparative studies: treatment dosage, therapists' competence or expertise, and an adequate realisation of the intended treatment concept. The expert ratings were then compared with an evaluation of an assessment of a research team from the University of Ulm and with the one from the Bernese research team. Contrary to the Bernese meta-analysis, both the experts and the Ulm research group conclude that only 5 or 8 of the 22 studies, respectively, could be accepted for a relatively fair comparison between the treatments under study. Out of the 5/8 studies, none could be considered fully suitable for a treatment comparison, at the most only moderately suitable. These remaining 5 or 8 studies, respectively, do not prove superiority of one treatment over the other. Hence, the "Dodo Bird Verdict" stands up under scrutiny. PMID- 9857720 TI - [From diagnosis to group therapy. A professional comment on Tschuschke et al.: Psychotherapy research--how it should (not) be done]. PMID- 9857721 TI - [Report of a self-analytic group discussion]. PMID- 9857722 TI - [Is early differentiation between psychogenic and somatogenic lumbago-ischialgia syndrome patients possible by anamnesis? Results of a screening study]. AB - Low back pain (LBP) is one of the most frequent causes for early retirement, hospital treatment and loss of days of work (Raspe u. Kohlmann 1993). Further differentiation and earlier diagnosis of psychogenic LBP could significantly reduce health costs. We interviewed 101 patients with LBP in the departments of Neurology and Neurosurgery (Gemeinschaftskrankenhaus Herdecke, University Witten/Herdecke) with the "structured biographical interview for patients with pain" (Egle 1992). This questionnaire has been used successfully differentiating between patients with organic and psychogenic chronic pain syndromes. It is a semi-standardised instrument based on observations, mainly made by Engel (1959), showing that many patients suffering from chronic benign pain syndromes had experienced psychic traumatisation during childhood. We compared a group of patients with clear neurological deficits and organic pain origin (n = 47) with a group of patients with psychogenic LBP (n = 25). Parameters were among others their biographical family constellations, their past medical history (also looking for symptoms of dissociation [conversion]), their present life and their coping with pain. In comparison with other studies examining similar aspects in patients who had pain in other parts of the body (Egle et al. 1991), our groups showed less discriminating results concerning biographical aspects. We found that also 40% of patients with organic pain origin hat traumatic constellations in their childhood. The following factors differentiated best between the both groups and characterised the psychogenic group: Intensity of pain was judged to be significantly higher with the "visual analog scale": age of the patient less than 40 years; distraction does not alter the pain; preceding experiences with physicians had often been negative; pain leads to impairment of familiar contacts; conflicts with parents during childhood could not adequately discussed with them at that time. These seven aspects can very well be part of a clinical history taking in a general medical setting, so that patients with LBP can be differentiated more effectively. PMID- 9857723 TI - [Organ donation from the viewpoint of the medical students]. AB - A questionnaire survey was carried out to examine the attitudes of 125 medical students, aged 19-37 years, toward organ donation. 73 of them were in their first semester and 52 senior students in their last year of the study. A return rate of 88% (senior students: 58%) was achieved. Although 59% (71%) expressed willingness to donate their organs, only 30% (50%) had signed an organ donor card. Concerns regarding definition and declaration of death, benefit of organ donation and feelings of the donor's family were identified in 51% (38%) of the students. 71% (79%) had already discussed this issue with their families. In summary, results of the study indicate that more intensified interdisciplinary discussion and information during the study of medicine could bring about an even more positive attitude toward organ donation. PMID- 9857740 TI - New insights in antibradycardia pacing. PMID- 9857741 TI - Radiofrequency catheter ablation of atrio-ventricular accessory pathways. AB - Radiofrequency catheter ablation of accessory pathways may represent the treatment of choice for patients with symptomatic atrio-ventricular reentrant tachycardia or atrial fibrillation and a high ventricular response through the accessory pathways with short antegrade refractory period. Moreover, catheter ablation has contributed towards understanding anatomical and electrophysiological characteristics of accessory pathways. As far as anatomical characteristics accessory pathways may be classified into fibers with proximal insertion in right or left atrium and fibers with proximal insertion in the atrio ventricular nodal junction. As far as electrophysiological properties are concerned, accessory pathways may be differentiated into non-decremental and decremental. In non-decremental by-pass tracts (the so-called Kent bundles) the impulse can be conducted antegradely, retrogradely and in both directions. Decremental accessory pathways generally show only antegrade conduction properties (the so-called Mahaim fibers) and can be classified in long and short pathways; in particular, catheter ablation has given considerable evidence that the vast majority (but not all) of them has a proximal insertion on the tricuspid anulus (atrio-ventricular pathways) or on the atrio-ventricular node (nodo ventricular pathways). Decremental accessory pathways with only retrograde conduction properties are involved in the so called "Permanent Junctional Reciprocating Reentrant Tachycardia". PMID- 9857742 TI - The implantable cardioverter defibrillator. PMID- 9857743 TI - Gene specific therapy for cardiac disease: the case of long QT syndrome. AB - Molecular genetics is progressively entering clinical practice. This new approach is modifying medical thinking as it becomes possible to diagnose diseases in their presymptomatic phase. It is therefore important for physicians to become acquainted with the "language" and the "methodology" of molecular biologists in order to balance opposite attitudes of the novice, i.e. skepticism and over expectation, and to establish a fruitful interaction with the molecular diagnostic laboratories. Long QT syndrome is an inherited disease that few years ago was still called "idiopathic" as the underlying causes were unknown. Clinicians are now becoming aware that what was considered as one disease is actually the common phenotype of defects in at least five different LQT-related genes and that therefore clinical heterogeneity is likely to parallel genetic heterogeneity. The first steps have been undertaken to define the relative prevalence of the molecular variants of LQTS, to develop gene-specific therapy and to perform risk stratification based on the molecular defect. In this article, current knowledge of the molecular bases of LQTS is reviewed and criteria are proposed to help defining 1) when it is appropriate to attempt molecular diagnosis, 2) how to interpret results of the diagnostic laboratory and 3) how molecular diagnosis may affect patients management. PMID- 9857744 TI - Stress echocardiography. PMID- 9857745 TI - Three dimensional echocardiography. PMID- 9857746 TI - Tissue Doppler imaging. PMID- 9857747 TI - New techniques in interventional cardiology. PMID- 9857748 TI - Interventional cardiology in the treatment of cardiogenic shock. PMID- 9857749 TI - Non-invasive assessment of the results of interventional cardiology. The value of radionuclide imaging techniques. PMID- 9857750 TI - [Image of the month. Uterine embolization of a persistent trophoblastic tumor]. PMID- 9857752 TI - [Clinical case of the month. Metastatic endocarditis: clinical observation and review of the literature]. AB - The authors describe a case of metastatic endocarditis associated with a gastric carcinoma. The diagnosis was made early and the treatment by surgery and chemotherapy allowed a survival of 18 months, which is unusually long. The differential diagnosis is discussed and includes nonbacterial thrombotic endocarditis, infectious endocarditis and primary tumors of the heart. PMID- 9857751 TI - [Pharma clinics. How I treat...or preventing pain with "patches"]. AB - There exists two distinct types of patches aiming at increasing the pain threshold either systematically or locally. The first type of analgesic patch is a real transdermal delivery system releasing fentanyl. Such potent opioid drug exerts a systemic effect. The other type of patch associates lidocain and prilocain to induce a local anesthesia of the skin without inducing a systemic effect. PMID- 9857753 TI - [Anatomo-clinical correlation. A case of invasive pulmonary aspergillosis]. AB - This report describes a case of invasive pulmonary aspergillosis affecting a young female patient who had benefited from a liver transplantation for a fulminant hepatic failure. Infections following liver transplantation can happen but in this case the short delay to the fatal outcome is unusual (6 days). Corticotherapy given prior to transplantation could have impaired the immune system. The authors discuss the infections due to Aspergillus species and particularly invasive pulmonary aspergillosis. PMID- 9857754 TI - [Viral implications in the etiopathogenesis of type 1 diabetes: current update]. AB - Type 1 diabetes mellitus results from autoimmune destruction of pancreatic beta cells. After having described genetic, immunological and metabolic factors, some researchers have hypothezised that environmental factors might trigger the autoimmune process. Based on epidemiological, anatomoclinical and animal studies, they suggest a role for virus infections. PMID- 9857755 TI - [Current classification of panniculitis]. AB - The inflammatory diseases of the hypodermis represent a difficult problem both for the accurate diagnosis and treatment. Several distinct etiologies and pathomechanisms can be responsible for such lesions. We present an updated panniculitis classification. PMID- 9857756 TI - [Tetany, spasmophilia, hyperventilation syndrome: theoretical and therapeutic synthesis]. AB - The hyperventilation syndrome (HVS), characterised by multiple somatic symptoms induced by inappropriate hyperventilation, constitutes the physiopathological manifestation of a common disorder in general medicine. As a synonym of spasmophilia or tetany, it has the advantage of offering diagnostic criteria, even though the latter are still vaguely defined. But its definition allows for objective measurements: indeed, a decrease in PCO2 during a hyperventilation provoking test and an abnormally low PCO2 rate at rest can be easily quantified. Moreover, the HVS concept offers a treatment which is both structured (respiratory reeducation, psychotherapy and pharmacology) and efficient. Yet, a number of scientific uncertainties still exist. There is no general agreement regarding the criteria which should be taken into account in a hyperventilation provoking test in order to diagnose an hyperventilation syndrome; the specificity of such a test is weak and a placebo can induce as many symptoms as can a HVS. Respiratory reeducation has good results but does not necessarily have an effect on PCO2. Some therapists see in it no more than a mechanism of relaxation and a rational explanation of frightening symptoms. This has led some authors to reject the term "hyperventilation syndrome" and to prefer the expression "chronic hyperventilation of unknown origin". PMID- 9857757 TI - [Troponin, a new myocardial infarction marker]. AB - The troponin (Tn) complex consists of three protein subunits referred to as TnT, TnI and TnC. Myocardium contains TnI and TnT isoforms which are not present in skeletal muscles and which can be separated from the muscular isoforms by immunological techniques. Using commercially available immunoassays, clinical laboratories are able to determine cardiac TnT and TnI (cTnT and cTnI) as quickly and reliably as classical cardiac markers. After acute myocardial infarction, cTnT and cTnI concentrations start to increase in serum in a rather similar way than CK-MB, but return to normal after longer periods of time (approximately one week). Because of their excellent cardiac specificity, Tn subunits appear ideally suited for the differential diagnosis of myocardial and muscular damage, for example in non cardiac surgery patients, in patients with muscular trauma or with chronic muscular diseases, or after intensive physical exercise. cTnT and cTnI may also be used for detecting evidence of minor myocardial damage, and therefore may find new applications in the management of patients with unstable angina. PMID- 9857758 TI - [Vaccination strategies for the general practitioner. II. New vaccines]. AB - In those vaccinal strategies of research action, we have to emphasize the practical data concerning the vaccination of the adolescents and adults according to the real situation estimated by the general practitioner. He has the capital preventive action to follow his patient's vaccination. PMID- 9857759 TI - [How I investigate...the age of adolescents using panoramic radiography]. AB - Stage of dental development gives an assessment of young adults chronological age. This is used in forensic medicine when birth data are lacking or doubted. The following method improves the accuracy of age determination in comparison with the commonly used method of Nolla. PMID- 9857760 TI - [Pharma-clinics. Drug of the month. Premelle (conjugated estrogens + medroxyprogesterone]. AB - Premelle, commercialised by Wyeth-Lederle, is a combination of conjugated estrogens 0.625 mg and medroxyprogesterone acetate 5 mg which is indicated in the treatment of menopause-associated problems, among which vasomotor symptoms, atrophic vaginitis and/or urethritis, and in the prevention and treatment of post menopausal osteoporosis. It is presented in two formulations, Premelle cyclic 5 and Premelle 5. The former, in which the progestagen is only given during the last 14 out of 28 days of the treatment cycle, is accompanied by regular bleeding and thus preferably indicated during perimenopause whereas the latter, in which the progestagen is given continuously and results in amenorrhea, is mostly indicated after menopause in order to improve long-term compliance. PMID- 9857762 TI - [What is your diagnosis? Iron deficiency anemia in malnutrition]. PMID- 9857763 TI - [Cholestasis: diagnosis]. AB - Cholestasis may present as extrahepatic (obstructive jaundice), as purely intrahepatic (hepatocellular), as part of a cholestatic hepatitis or as a result of intrahepatic obstruction. Anamnestic clues and clinico-chemical analyses (bilirubin, gamma-GT and alkaline phosphatase) together with sonography and finally ERCP usually will furnish the diagnosis. However, with technical advances computed tomography and MRI cholangiography will soon emerge as often useful techniques in hepatobiliary imaging. PMID- 9857764 TI - [Cholestasis: drug therapy]. AB - For the medical treatment of cholestasis plant alkaloids, phenobarbital and S adenosyl-L-methionine (SAMe) have been used. Although the mode of action of these substances is understood in part, the treatment of patients was unsuccessful. In contrast, ursodeoxycholic acid, a physiologically occurring bile acid in man, was successful. The daily dosage of ursodeoxycholic acid is 10-15 mg/kg bodyweight. Best results have been obtained in primary biliary cirrhosis where symptoms improved markedly, the cholestasis-indicating enzymes and immunoglobulin M decreased significantly. After long-term therapy even liver histology improved. Recently it has been shown that ursodeoxycholic acid prolongs the interval to liver transplantation. Ursodeoxycholic acid has to be taken lifelong, because interruption of therapy, even after long periods of continued treatment will induce a rebound of cholestasis. Ursodeoxycholic acid therapy is without side effects. In patients with primary sclerosing cholangitis ursodeoxycholic acid prolongs life expectancy only in combination with endoscopic bile duct dilatation. PMID- 9857765 TI - [Cholestasis: interventional therapy]. AB - Treatment of cholestasis by bile duct obstruction is nowadays dominated by endoscopy and other interventional measures. Intraductal concrements can be removed almost always without open surgery. This is as a rule achieved by a transpapillary approach with the additional use of lithothripsy. Percutaneous transhepatic treatment of concrements is restricted to patients in which a transpapillary approach is impossible. Stenoses of biliary ducts by malignancies are treated with plastic or metallic endoprosthesis if curative resection is precluded. Percutaneous drainage is a valid option in reserve. Stenosis of the papilla by scars, tumors or sphincter dysfunction are mainly treated endoscopically, malignancies--if possible--by radical surgery. Rare causes for obstruction have also to be considered. They are in most instances amenable to interventional procedures. PMID- 9857766 TI - [Diagnosis of chronic pancreatitis]. AB - Chronic pancreatitis is typically characterized by clinical (abdominal pain, steatorrhea, loss of body weight), morphological (calcifications, dilated ductus pancreaticus) as well as functional (maldigestion, diabetes mellitus) parameters. Since the diagnosis of chronic pancreatitis is hampered by the inavailability of early histological confirmation, it is therefore based on morphological (ultrasound, ERP, EUS, CT) and functional (faecal elastase) criteria. Due to the poor correlation between morphological and functional parameters in the early phase of the disease, both are complementary at this stage. While the diagnosis of severe cases of chronic pancreatitis with steatorrhea is hardly a challenge in clinical practice, the differential diagnostic evaluation of mild and moderate cases remains a major clinical problem. ERP remains to be the most sensitive morphological procedure, while determination of faecal elastase is the most sensitive and specific "tubeless" pancreatic function test available today and in the future prove to be rapid, easy to handle and highly practicable in clinical routine. PMID- 9857767 TI - [Chronic pancreatitis: nutrition and pain therapy]. AB - Therapy of chronic pancreatitis rests on five arms: Avoidance of alcohol, treatment of pain, replacement therapy for exocrine and endocrine insufficiency and adequate nutrition. Alcohol withdrawal improves pain and the patient's compliance. It also seems to retard the chronic inflammatory process. Therapy of pain depends on the pathomechanism of pain. There is a lack of prospective, controlled studies comparing various treatment regimens. Thus, treatment options are partly dependent on the experience of the physician taking care of the patient and include i.e. for pseudocysts: surgical vs percutaneous or endoscopic drainage; for stenosis of the main pancreatic duct close to the papilla: surgical vs endoscopic drainage (stents); for distal bile duct stenosis: endoscopic stents vs biliodigestive anastomosis vs pancreatic head resection; for pancreatic stones: extracorporal shock wave lithotripsy followed by endoscopic stone extraction vs surgery (pancreaticojejunostomy), finally for inflammatory tumor of the pancreatic head combined with pain with or without compression of the distal bile duct or duodenum: duodenum-preserving pancreatic head resection vs Whipple resection. Patients with pain resistant to medical treatment may be candidates for a transcutaneous blockade of the plexus coeliacus or for epidural nerve blockade before one choses a surgical procedure. Application of pancreatic enzymes does not seem to have a major beneficial effect on pancreatic pain. Modification of nutrition has become less restrictive. Thanks to improved substitution with acid resistant porcine pancreatic extracts with high lipase activity, fat restriction is no longer of paramount importance. However, supply with sufficient calories is still difficult due to pain, inadequate compliance and hypermetabolism. PMID- 9857768 TI - [Pancreaticolithiasis and pancreatic pseudocysts--interventional therapy]. AB - Interventional (non-surgical) therapy is a relative new approach in the management of chronic pancreatitis. Indications for endoscopic therapy are pseudocysts and strictures or stones of the main pancreatic duct. Endoscopic sphincterotomy, extraction of pancreatic duct stones, extracorporal shock-wave lithotripsy (ESWL), pancreatic duct stenting and drainage of pseudocysts are all methods with a high success rate but with a low complication rate. Therefore in patients with chronic pancreatitis and with a failure of conservative management interventional techniques have to be considered. They may be an alternative to surgical procedures. PMID- 9857769 TI - [Patient with right-sided epigastric pain and "negative" ultrasound]. AB - In most cases pain in the right upper abdominal quadrant is of biliary origin. Diseases of the biliary system are often visualized by ultrasonography; however, a negative test does not rule out a biliary cause. Moreover, a number of several non-biliary diseases have to be considered. Patient history, physical examination and laboratory findings determine the further diagnostic approach and include radiologic (i.e. CT/scintigraphy) and/or endoscopic (upper GI endoscopy, ERCP) examinations. In difficult diagnostic situations specialized diagnostic tools may be indicated (e.g. manometry in suspected dysfunction sphincter of the Oddi. PMID- 9857770 TI - [Incidental ultrasound findings of the gallbladder]. AB - The low threshold for sonographic examinations of the gallbladder, the relative high incidence of gall bladder disease and the extraordinary quality of ultrasound imaging leads to a remarkable number of chance findings. The image quality of new ultrasound scanners allows in most cases a rapid diagnostic decision. Therapeutic consequences depend in the first line on the following carefully directed anamnesis. The most common findings are gallstones, cholesterinosis, polyps and sludge. Rare but important findings are porcelain gallbladder, adenomyomatosis and cysts of the common bile duct. PMID- 9857771 TI - [The patient with isolated pancreatic enzyme elevation]. AB - Elevated serum lipase and amylase activities are the most common laboratory tests for pancreatitis. Next to pancreatic pathologies elevated pancreatic enzymes are associated with several other diseases. If no such association is found, an idiopathic form of pancreatic enzyme elevation can be diagnosed. For this entity no pancreatic pathologies have been found. In contrast to other diseases with elevated pancreatic enzymes, the idiopathic form needs neither further diagnostic management nor a regular follow-up. PMID- 9857773 TI - Mental disorder and violence: results of epidemiological studies in the era of de institutionalization. PMID- 9857774 TI - The growing belief that people with mental illnesses are violent: the role of the dangerousness criterion for civil commitment. AB - In response to a controversy concerning whether the stigma of mental illness has declined significantly in the United States in the past several decades, we assessed changes in public perceptions that mentally ill people are violent. Specifically, we compared answers to an open-ended question regarding respondents' understanding of the term "mental illness" from two nationally representative surveys, one conducted in 1950 and one in 1996. In an earlier paper, we reported the finding that perceptions of violence not only failed to decrease but actually increased significantly between 1950 and 1996. In this paper, we explore the possibility that the dangerousness criterion for involuntary commitment, widely adopted in the United States beginning in the 1960s, has contributed to the unexpected increase in perceptions that mentally ill people are dangerous. We find that, among respondents who mention violence in their description of a mentally ill person, the percentage who use "dangerous to self or others" phrasing to indicate this belief increased substantially, from 4.2% in 1950 to 44.0% in 1996. Moreover, eliminating these respondents from consideration, there was a slight decrease in perceptions of violence between 1950 and 1996. We discuss the possibility that the adoption of the dangerousness criterion, which was intended to protect the civil liberties of mentally ill persons, may also have had the unintended consequence of increasing the stigma of mental illness in the United States. PMID- 9857775 TI - The psychiatric epidemiology of violent behaviour. AB - This paper reviews the current state of the debate on the relationship between mental disorder and violent behaviour. Starting from the discussion of methodological approaches to assessing a possible association, the most important studies carried out on the issue in recent years are discussed. Their results concur in supporting the assumption that there is a moderate but reliable association between mental disorder and violence. However, this does not imply that people with mental illness are generally more likely to commit violent acts than members of the general population. An elevated risk of violent behaviour is only evident for specific psychiatric diagnoses and symptom constellations. For schizophrenia and other psychotic disorders, a significant increase in the likelihood to commit violent acts is reported. Substance use disorders and antisocial personality disorder, however, represent a markedly higher risk for violent behaviour. The article further discusses possible determinants of violent behaviour such as psychotic symptoms and comorbidity with substance abuse and considers who is at particular risk of becoming a target of violent acts. PMID- 9857776 TI - The Camberwell Study of Crime and Schizophrenia. AB - I report a population-based longitudinal study of all incident cases of schizophrenia (n = 538) in the London Borough of Camberwell between 1964 and 1984. The rates of criminal convictions are compared with those in a control sample representative of non-schizophrenic mental disorders matched for age, sex and period. The results show that women with schizophrenia have an increased rate of offending across all offence categories (rate ratio = 3.3). There is no overall increase for males, but a specific increase for violent convictions (rate ratio = 3.8). Looking at the risk of acquiring a first conviction, there is an independent but modest effect of schizophrenia (hazard ratio = 1.4), but the effects of gender, substance abuse, ethnicity and age at onset were more substantial. PMID- 9857777 TI - Epidemiological investigations of the associations between major mental disorders and crime: methodological limitations and validity of the conclusions. AB - Five epidemiological investigations of post-World War II birth cohorts have examined the relation between major mental disorders and criminality. This article critically reviews the objectives, methodology and findings of these investigations. The validity of the results is assessed as are the limitations which restrict their interpretation. It is concluded that persons who develop major mental disorders are at increased risk across the lifespan of committing both non-violent and violent crimes. However, this increased risk may be limited to generations of persons with major mental disorders born in the late 1940s, 1950s and 1960s. It is hypothesized that these generations of persons with major mental disorders have received inappropriate and inadequate mental health care and that they include a larger proportion of individuals than previous and perhaps subsequent generations, who display a pattern of stable antisocial behaviour from childhood through adulthood. PMID- 9857778 TI - Understanding causal paths between mental illness and violence. AB - The stigma associated with mental illness is a major concern for patients, families, and providers of health services. One reason for the stigmatization of the mentally ill is the public perception that they are violent and dangerous. Although, traditionally, mental health advocates have argued against this public belief, a recent body of research evidence suggests that patients who suffer from serious mental conditions are more prone to violent behaviour than persons who are not mentally ill. It is a point of contention, however, whether the relationship between mental illness and violence is only one of association, or one of causality; that mental illness causes violence. A proven causal association between mental illness and violence will have major consequences for the mentally ill and major implications for caregivers, communities, and legislators. This paper outlines the key methodological barriers precluding casual inferences at this time. The authors suggest that a casual inference about mental illness and violence may yet be hasty. Because a premature statement advocating a causal relationship between mental illness and violence could increase stigma and have devastating effects on the mentally ill the authors urge researchers to consider the damage that may be produced as a result of poorly substantiated causal inferences. PMID- 9857780 TI - Psychotic symptoms and violent behaviors: probing the components of "threat/control-override" symptoms. AB - Evidence for an association between mental illness and violence has grown in recent years, leading many to ask why such an association exists. One hypothesis links elevated rates of violence among people with mental illness to a small set of psychotic symptoms--so called threat/control-override (TCO) symptoms. Several studies have supported this hypothesis, but none has examined which of the components, threat or control-override--if either--predominates in explaining violence. To explore this issue we used data from a two-stage epidemiological study (n = 2741) conducted in Israel. Data on TCO symptoms were collected using two methods--fixed-format self-report questions from the first stage and psychiatrists' ratings based on interviews using the Schedule for Affective Disorders and Schizophrenia (SADS) from the second. Results show that both a measure of threat and a measure of control-override are independently associated with violent behaviors. Results also show that neither method--neither fixed format questions nor psychiatrist rating--predominates in explaining violence. In sum, these results indicate that both the threat and the control-override components of the TCO concept are useful in predicting violent behaviors and that a better measurement of the TCO concept is achieved using a multimethod approach. PMID- 9857781 TI - Gender differences in the relationship between mental illness and violence: evidence from a community-based epidemiological study in Israel. AB - Although males are generally more likely than females to report violent behaviors, emerging evidence suggests that the gender gap is substantially reduced among individuals with mental illness. This paper investigates whether the associations between gender and self-reported violent behaviors (fighting and weapon use) are moderated by three mental health indicators--treatment status, psychiatric diagnosis, and threat/control-override psychotic symptoms. Data from a two-stage epidemiological study conducted in Israel (weighted N = 2706) are analyzed using descriptive statistics and logistic regression analysis. Results indicate that treatment status and psychiatric diagnosis moderate the association between gender and fighting, but leave open questions both about the moderating role of threat/control-override symptoms and about the implications of mental illness for the gender/weapon use relationship. PMID- 9857779 TI - When symptoms of psychosis drive serious violence. AB - Three studies are briefly reviewed as contributions to the epidemiology of violence and mental disorder. The samples were severally drawn from a then substantial regional pre-trial prison, a national high security hospital service and a general psychiatric hospital with a mainly local clientele. Delusions emerged consistently as having an important role in the precipitation of a violent act by a person with a psychotic illness; the more serious the act, the more delusions appear to have had a direct role. Nevertheless, given that delusions are very common symptoms of psychosis, but serious violence by someone with psychosis is unusual, other mediating factors must be operating. There is some support each for an interactional effect with other symptoms of the psychotic process, an influential effect on further symptom,-particularly affective symptom-development and at least two social relationship mechanisms. This augurs well for a shift from hitherto primarily observational work to hypothesis-generated research. PMID- 9857782 TI - Male-female differences in the setting and construction of violence among people with severe mental illness. AB - Data from a sample of severely and persistently mentally ill involuntary patients indicated that differences in violence between males and females in the 4 months prior to hospital admission depended on the measure. In the bivariate analysis, males had a greater prevalence of violence on the two indicators which separated more serious violence from lesser and no violence; but there was no gender difference on the more inclusive measure which incorporated threats and fights not involving weapons or injuries. In multivariate analysis when other relevant predictors were controlled, gender was significant in predicting only the most inclusive indicator of violence and only in interaction with substance abuse problems. PMID- 9857783 TI - Taking the wrong drugs: the role of substance abuse and medication noncompliance in violence among severely mentally ill individuals. AB - Increasing numbers of severely mentally ill individuals are being treated in nonhospital, community-based settings and public concern about potential violence by these individuals has increased, often as a result of tragic, albeit uncommon events. The present study examines potential predictors of serious violence among persons with severe mental illness (SMI), with a specific focus on the joint effect of substance abuse and medication noncompliance. Subjects in the study are involuntarily admitted inpatients with SMI awaiting a period of court-ordered outpatient treatment, termed "involuntary outpatient commitment". During enrollment in a longitudinal outcome study of the effectiveness of OPC, 331 subjects and, whenever feasible, family members or other informants were interviewed. In addition, complementary data were gathered by review of involuntary commitment records and hospital records. Data collection included sociodemographic characteristics, illness history, clinical status, medication adherence, substance abuse and violent behavior during the 4 months preceding hospitalization. Descriptive and multivariable logistic regression procedures were used to examine the association between serious violent acts and a number of personal, social, and clinical characteristics. The combination of medication noncompliance and substance abuse was a significant predictor of serious violent acts in the community. Individuals who had problems with both alcohol and illicit drug abuse appear to be at greatest risk for violence. These results suggest that reducing violence risk among persons with SMI requires an aggressive approach to improving medication adherence in the context of integrated mental health and substance abuse treatment. PMID- 9857784 TI - Mental illness and criminal violence. AB - This article examines the relationship between criminal violence and mental illness. Our data suggest that mentally ill persons tend to have an increased risk for committing violent offenses, and that the violent offending by these individuals tends to be recidivistic. Our findings suggest that parents who have both committed violent offenses and experienced a psychiatric hospitalization increase the risk of violent offending among their offspring. We propose the hypothesis that mentally ill parents transmit a biological characteristic which may genetically predispose their child towards criminal violence. Prenatal disturbances during critical periods of fetal development may provide clues regarding the etiology of criminal violence. PMID- 9857785 TI - Psychiatric impairment, social contact, and violent behavior: evidence from a study of outpatient-committed persons with severe mental disorder. AB - The need to better understand and manage risk of violent behavior among people with severe mental illness in community care settings is increasingly being recognized, as public-sector mental health systems face mandates to provide more cost-effective services in less restrictive environments. The potential for serious violence in a small proportion of severely mentally ill (SMI) individuals has emerged as a key factor that increases cost and limits continuity and normalization of community-based services for populations with psychiatric disabilities. A major challenge to developing better strategies for risk assessment and management in community care settings involves specifying complex interactions between psychiatric impairment and the conditions of social life- including the quality and frequency of contact with others at close quarters. This is a study of the determinants of violent behavior in a sample of 331 adults with severe mental disorders in community-based treatment. An interaction between severity of functional impairment and frequency of social contact was found to be significantly associated with risk of violence. Among respondents with Global Assessment of Functioning (GAF) scores in the lowest 20%, more frequent contact with family and friends was linked to a higher probability of violent events. However, among better functioning respondents, frequent social contact was associated with lower risk of violence and greater satisfaction with relationships. These findings suggest that, where violence risk is concerned, the most salient feature of psychiatric impairment is the impairment of social relationships--the ways in which disorders of thought and mood not only distort one's subjective appraisal of experience and threat, but impair the ability to relate meaningfully to others, to resolve conflict and derive necessary support from family and friends. Thus, social contact may be a mixed blessing for SMI individuals. For some, it signals a positive quality of life, but for others- particularly those with extreme psychiatric impairment--frequent contact may add to conflict, stress, and increased potential and opportunity for physical violence. The impact of psychiatric impairment on violent behavior cannot be known in isolation, but must be considered in a social context. Effective community-based strategies to anticipate and prevent violence in the lives of persons with severe mental illness must take into account such interactions between social and clinical variables. PMID- 9857786 TI - Risk reconsidered: targets of violence in the social networks of people with serious psychiatric disorders. AB - Risk Reconsidered: Targets of violence in the Social Networks of People with Serious Mental illness. This exploratory analysis addresses the questions: 1) Who among the members of the social network of a person diagnosed with a major psychiatric disorder is likely to become a target of violence? 2) What kind of relationships do targets have with respondents in terms of the quality and quantity of interactions?, and 3) What are the risk factors that contribute to being a target of violence for people who are in the social networks of persons with serious mental disorders? The samples of 169 people with serious mental disorders were followed for 30 months. A logistic regression model of the risk for being a target of violence among the members of the cohort's social network reveals that both target and respondent characteristics are salient, and that mothers who live with adult children who have schizophrenia and co-occurring substance abuse bear a substantially elevated risk of becoming a target of violence, compared to other social network members. Other factors that elevate the risk for being a target of violence are being an immediate family members of the respondent, more time in residence with the respondent, and whether the respondent is financially dependent on the family. Respondents with the most mental health center visits had lower odds of committing an act or threat of violence against a social network member. PMID- 9857787 TI - Suicide among personality-disordered offenders: a follow-up study of 1943 male criminal offenders. AB - Suicide mortality among all male criminal offenders in Sweden who had been subjected to a major forensic psychiatric examination 1988-1991 (n = 1943) was studied, with special reference to offenders with personality disorders. The cohort was followed until the end of 1995. Altogether 135 individuals (6.9%) died during the follow-up period; the mode of death was suicide in 50 individuals (2.6%). The unadjusted suicide mortality ranged from 2.8% among those with personality disorders to 6.1% among those with drug-related psychosis. The standardised mortality ratio (SMR) among personality-disordered offenders was 1212, i.e. around 12 times that of the general population. Survival analyses by means of Cox regression models were performed to identify background factors associated with completed suicide. No specific principal diagnosis showed significantly increased risk for completed suicide. However, concomitant depression and drug abuse were significantly linked to suicide. Violent crime showed no association. Among personality-disordered offenders suicide methods did not differ from those of suicide victims in the general population. There was no association between violent index criminality or between life-time violent criminality and choice of a violent suicide method. PMID- 9857788 TI - Clinical prediction of violence as a conditional judgment. AB - Previous research on the prediction of violence in mentally ill individuals has focused primarily on determinations about the appropriateness of institutional confinement. The assessment and management of violent, mentally ill individuals in the community, however, requires clinicians to take a more detailed look at the factors that might precipitate or inhibit violence in the community. This paper examines a model of conditional prediction, in which clinicians provide assessments of the factors that they expect to be associated with violence in particular patients. These types of predictions were elicited from clinicians for a sample of 712 patients seen in an urban psychiatric emergency room. These patients were then followed in the community for 6 months, using both interviews and official records. Results showed that clinicians were generally accurate about the seriousness and location of the violence, but overestimated the role of medication compliance and drug use in the violent incidents. PMID- 9857789 TI - On the State of the Public Health: the Annual Report of the Chief Medical Officer of the Department of Health of England, 1995: an example of increased visibility of mental health at a national level. PMID- 9857790 TI - The Netherlands Mental Health Survey and Incidence Study (NEMESIS): objectives and design. AB - The article describes the objectives and design of a prospective study of the prevalence, incidence and course of psychiatric disorders in a representative sample of non-institutionalized Dutch adults. A total of 7146 men and women aged 18-64, contacted through a multistage sample of municipalities and households, were interviewed at home in 1996. The primary diagnostic instrument was the CIDI, which determines the lifetime occurrence of DSM-III-R disorders. The disorders included were: mood disorders, anxiety disorders, eating disorders, schizophrenia and other non-affective psychotic disorders, and dependence and abuse of psychoactive substances. Follow-up measurements in the same sample were scheduled at 12 and 36 months. The net response to the first measurement was 69.7%. Poststratification weightings were applied for gender, age, marital status and degree of urbanization. Limitations and advantages of the study design are discussed. Findings are reported elsewhere in this issue. PMID- 9857791 TI - Prevalence of psychiatric disorder in the general population: results of The Netherlands Mental Health Survey and Incidence Study (NEMESIS). AB - This article reports the initial results of a prospective study on the prevalence of psychiatric disorders in the Dutch population aged 18-64. The objectives and the design of the study are described elsewhere in this issue. A total of 7076 people were interviewed in person in 1996. The presence of the following disorders was determined by means of the CIDI: mood disorders, anxiety disorders, eating disorders, schizophrenia and other non-affective psychoses, and substance use disorders. Psychiatric disorders were found to be quite common. Some 41.2% of the adult population under 65 had experienced at least one DSM-III-R disorder in their lifetime, among them 23.3% within the preceding year. No gender differences were found in overall morbidity. Depression, anxiety, and alcohol abuse and dependence were most prevalent, and there was a high degree of comorbidity between them. The prevalence rate encountered for schizophrenia was lower (0.4% lifetime) than generally presumed. A comparison with findings from other countries is made. Relevant determinants of psychiatric morbidity were analysed. PMID- 9857792 TI - Risk factors for onset and persistence of psychosis. AB - Clinical practice, training and evaluation of treatment in the functional psychoses continues to be carried out mostly along the traditional line of separation by diagnostic entity. However, the combined evidence from research on risk factors for onset and for persistence of psychotic illness indicates quantitative, but not qualitative, differences between categories of schizophrenia and affective psychosis. "Developmental" factors, such as childhood dysfunction, increased cerebral ventricle size and familial morbid risk of schizophrenia operate preferentially, though not specifically, at that end of the psychopathological spectrum characterised by a preponderance of negative features. On the other hand, "social" factors, such as ethnic group, adverse life events and familial morbid risk of affective disorder have a larger impact at the end associated with predominance of affective features. Heterogeneity in the functional psychoses may thus be best conceived as two discrete effects operating at different ends of a continuous psychopathological spectrum. The use of highly reliable but arbitrary diagnostic categories may introduce serious bias in aetiological and treatment research. Evidence supporting the validity of a model of shared risk factors for continuous characteristics needs to be further elaborated and incorporated into our concepts of psychotic illness. PMID- 9857793 TI - Shared social environment and psychiatric disorder: a multilevel analysis of individual and ecological effects. AB - We examined associations, in terms of relative and population-attributable risks, between shared social environment at the neighbourhood level and (1) treated incidence of non-psychotic, non-organic disorders, and (2) subsequent level of service consumption. The multilevel analysis used linked records of all individuals in contact for the first time with any catchment area mental health service for non-psychotic, non-organic disorder over various specified time periods between 1981 and 1995. Socioeconomic indicators of 36 neighbourhoods in the city of Maastricht yielded a multivariately defined neighbourhood deprivation score. There were significant linear trends in the association between level of deprivation and treated incidence, especially in the population under 35 years of age (adjusted rate ratio for linear trend 1.17, 95% confident interval 1.11 1.23), who constituted around half of the patient population. The fraction of the incidence of psychiatric disorder attributable to deprivation was 17.8%. Multilevel analyses of rates of a second cohort, with cases divided according to level of service use over a standardised follow-up of 5 years after first contact with psychiatric services, revealed that the effect of deprivation scores on rates declined with intensity of out-patient service use, but increased with level of in-patient use. Up to 50% of in-patient episodes in this group could be attributed to neighbourhood level of deprivation. The increase in risk conferred by neighbourhood deprivation remained after adjustment for the individual-level equivalent. The findings therefore suggest that elements in the shared social environment influence both incidence and severity of non-psychotic, non-organic disorders, over and above any individual-level effect. PMID- 9857794 TI - Sick-leave in women with alcohol dependence or abuse: effects of additional psychiatric disorders. AB - The study objective was to assess sick-leave among women investigated in a general population survey of alcohol dependence/abuse (ADA). A total of 399 women, selected by stratified random sampling, were interviewed and diagnosed according to DSM-III-R. Data on sick-leave were obtained by linkage with the Social Insurance records. The study found that women with ADA but without other psychiatric disorders had an increased number of annual sick-leave spells--1.82 compared with 1.47 in the reference population-whereas the mean duration was similar. Women with ADA and other psychiatric disorders had 2.38 annual spells, but also considerably longer spells (mean 16.54 days vs 9.70). Socio-economic differences were large, with the less privileged groups having both more and longer spells. Stepwise multiple regression showed that both ADA and other psychiatric disorders contributed to high sick-leave incidence and duration, as, to a lesser extent, did low education and low social group (the last variable only affected duration of the spells). The frequency of disability pension/long sickness spells was higher in women with ADA (odds ratio of 2.95). We concluded that there is a strong association between ADA and sick-leave, which increases considerably in the presence of additional psychiatric disorders. PMID- 9857795 TI - Assessing the feasibility of recruiting a randomly selected sample of psychiatrists to participate in a national practice-based research network. AB - This study tested the feasibility of recruiting and retaining a randomly selected sample of psychiatrists compared to a volunteer sample to participate in the American Psychiatric Association's Practice Research Network (PRN). One hundred forty psychiatrists were randomly selected and contacted by phone by peer psychiatrists for recruitment into the PRN. As a comparison group, a sample of 146 self-selected volunteer psychiatrists were also included in the study. Recruited and volunteer psychiatrists were asked to participate in three studies to assess study compliance and retention. The representativeness of each sample was evaluated by comparing the psychiatrists' sociodemographic and practice characteristics to existing national data on psychiatrists. Study response rates and long-term retention rates were compared for the two groups. Sixty-one percent of the recruited sample who were eligible to participate in the network were willing to participate. Both the recruited and volunteer samples were broadly representative of the American Psychiatric Association's membership (with some differences in race, ethnicity and board certification). Of the recruited sample, 74.5% (38/51) successfully completed the network's first three pilot studies compared to 72.5% (98/135) of the volunteer sample. No psychiatrists in the recruited sample withdrew from the network compared to 2.1% (3/138) of the volunteer sample. These findings indicate a randomly selected sample of psychiatrists can be recruited and retained to participate in practice-based research. These methods can be used to enhance the generalizability of observational health services research studies, which require the participation of practicing clinicians. More effective methods should be tested to enhance participation rates. PMID- 9857796 TI - The impact of work-related rehabilitation on the quality of life of patients with schizophrenia. AB - In a cross-sectional study of 60 outpatients with schizophrenia (mean age 36.3 +/ 11.1) the effect of a work-related rehabilitation programme on the patients quality of life was investigated. A group of patients with the same diagnosis, but without rehabilitation, served as controls. Patients of the rehabilitation group had been attending the programme for a mean duration of 15.0 months. The programme focuses on occupational and everyday skills and also involves social aspects. The patients' quality of life was assessed using two self-administered questionnaires: the Munich List of Life Dimensions (MLDL, Heinisch et al. 1991) and the Everyday Life Questionnaire (Bullinger et al. 1993). Life satisfaction as well as functional quality of life were higher in the rehabilitation group in the majority of domains assessed. Difference between groups was highest for satisfaction with work, followed by leisure-time activities, independence and friendships/acquaintances. Results indicate that the rehabilitation programme acts like a "lever" which, applied to one point (work, day structuring), subsequently affects most domains of daily living. PMID- 9857797 TI - [Norwegian physicians in war?]. PMID- 9857798 TI - [Who will shape the female body? Culture, surgery and conflict]. PMID- 9857799 TI - [A competence center for Sjogren's syndrome--an example to be followed]. PMID- 9857800 TI - [Intestinal invagination in children]. PMID- 9857801 TI - [Intestinal invagination in children in the county of Hordaland 1983-92]. AB - The incidence of intussusception was examined with respect to whether children lived in the city of Bergen, where pediatricians were the primary contacts in 45% of the cases, or in the remaining municipalities where pediatricians were the primary contacts in 18%. Diagnosis and course of illness were compared for children admitted by pediatricians and general practitioners. Information was retrieved from hospital medical records. During 1983-92, 142 children 0 to 14 years were given the diagnosis on basis of barium enema (139 children) or surgery (three children). The incidence was 1.8 per 1,000 children per year for Bergen vs. 1.4 in the rest of the county (p = 0.3). Children admitted by pediatricians more often had the correct referral diagnosis (70% vs. 42%, p = 0.003), but they did not differ with respect to median age (8 months for both), median duration of symptoms (12 vs. 24 hours), percent in need of surgery (58% vs. 59%) or in symptoms. The lack of difference in incidence between Bergen and the more distant municipalities suggests that spontanous resolution of intussusception is uncommon. Although pediatricians more often admitted the children with the correct diagnosis, duration of symptoms and outcome did not differ from those admitted by general practitioners. This suggests that the general practitioners also appreciated the urgency of the symptoms. PMID- 9857802 TI - [Increased incidence of thyroid cancer among Norwegian women married to fishermen]. AB - The relationship between thyroid cancer in women and the occupation of their spouses was examined in a retrospective cohort study. Of the 2.9 million women registered in the Central Population Registry of Norway by the end of 1991, 1.2 million had a spouse registered with an occupation in one or more of the censuses of 1960, 1970 or 1980. These women were included in the study. Based on the first digit of their husbands' five-digit Nordic occupational code, the women were assigned to ten broad categories. A standardised incidence ratio (SIR) and 95% confidence interval were calculated for each occupational category. The women were further subdivided and analysed in 71 groups defined by the first two digits of their husband's occupational code. Among the women included in the study, a total of 2,409 cases of thyroid cancer were reported to the Cancer Registry of Norway during the period 1960-92. A significantly elevated risk of thyroid cancer was found only among women whose spouses belonged to the occupational category Agriculture, forestry or fishery (n = 208,279), with a SIR of 1.13. In the subgroup Fishing, whaling and sealing work (n = 40,839), the risk was even higher with a standardised incidence ratio of 1.91. Our data support the proposed relationship between increased risk of thyroid cancer and mode of living, more specifically dietary fish or other seafood. PMID- 9857804 TI - [Quality assurance of psychiatric consultations in somatic hospitals]. AB - Research has shown great variety in the clinical practice of consultation-liaison psychiatry in and between different countries. This paper presents the Norwegian experiences from a European collaborative study of quality management in consultation-liaison psychiatry. We describe a dynamic model for total quality management based on regular registration of some clinical data and the subsequent feed-back on changes of these. We discuss our experiences with this model and obstacles met in everyday work. Finally we point to different ways of implementing this method on a broader base in consultation-liaison psychiatry. PMID- 9857805 TI - [Constrictive pericarditis illustrated by an unusual case report]. AB - Constrictive pericarditis is an uncommon condition. Previously, tuberculosis or other bacterial infections were prevalent causes, often with prominent pericardial calcification. Presently, many patients with constrictive pericarditis of other aetiologies have lesser degrees of structural changes in the pericardium. We report on a case with severe symptoms where the correct diagnosis was elusive because of absent or minimal preoperative pericardial pathology. The clinical, echocardiographic and haemodynamic features of constrictive pericarditis are reviewed. We recommend thorough echocardiographic evaluation of central haemodynamics in patients with symptoms of heart failure when the aetiology is not readily apparent (e.g. previous myocardial infarction dilated cardiomyopathy or valvular disease. PMID- 9857803 TI - [Magnetic tomography of the central nervous system in adults with myelomeningocele]. AB - The Norwegian training and counselling centre for patients with rare disabilities is a national project with the aim of developing new models for services designed to improve the quality of life of these patients. As at 1 July 1997, 130 adults with myelomeningocele were registered users of the Centre. This article is a retrospective review of reports from MRI evaluations of the central nervous system of 61 (40%) of the centre's users, performed in seven departments of radiology. Caput, the craniocervical region and distal parts of the medulla were the most frequently examined areas; the thoracal medulla was examined in half of the cases. There were considerable differences with respect to the structures examined. Enlargement of the lateral ventricles was as frequent in patient without shunt (68%) as in patients with shunt (53%). Dysmorphology of the corpus callosum was seen in 39%. Intracranial structural abnormalities were often described without a report on the intracranial pressure. Herniation of the cerebellar tonsils was reported in 66% of patients, tethering of the spinal cord in 90%, and syringomyelia in 13%. In most cases, MT uncovered conditions with implications for management. We suggest that standard routines should be developed for examinations of the central nervous system of patients with myelomeningocele. PMID- 9857806 TI - [Central stimulants in adults with AD/HD. Do they help?]. AB - This article is a review of five controlled studies of the efficacy of methylphenidate in adults with attention deficit/hyperactivity disorder (AD/HD). All five had a placebo cross-over design. In one of the studies, patients noted from the side-effects whether they were taking methylphenidate or placebo, and it is, in fact, unlikely that any of the studies were blind. The diagnostic criteria used in two of the studies were not appropriate according to present knowledge, leaving three samples of subjects meeting the present criteria for AD/HD. In one study, the patient material was extremely selective: 74% of subjects belonged to social class 1 and 2, and only 35% of those who came for treatment were eligible. Only one study has an acceptable (though not perfect) design. In this study of patients in a psychiatric clinic, no difference was found between methylphenidate and placebo. In two studies, subjects who improved on methylphenidate were followed up. Of the total of 24 subjects in these two samples, only four continued with methylphenidate after 3-12 months; two of them suffered from narcolepsy and one was a substance abuser. Thus, the efficacy of methylphenidate on AD/HD in adults has not been demonstrated. Present research is more against than in favour of its existence. PMID- 9857807 TI - [Central stimulants in adults with AD/HD. Can they be abused?]. AB - As early as in 1979, abuse was observed when adults with MBD were treated with psychostimulants. In a study from 1981, five patients (19%) had been stimulated or excited. It was concluded that there was no reason to believe that adults with MBD were immune from stimulant abuse problems. In subsequent studies, it has been the rule to exclude patients who abuse alcohol or drugs. Obviously, there has been agreement that treatment implies a considerable risk of abuse of stimulants. However, nobody has systematically studied the frequency of psychostimulant abuse in subjects with AD/HD with and without abuse of alcohol and drugs, neither during the study nor at follow-up. There has been some attempts at using pemoline, a stimulant which cannot be abused. However, the mental and physical complications have been severe. Some preliminary studies indicate that stimulants can be used to treat alcoholics and substance abusers with concurrent AD/HD. Before such treatment can be recommended, the observations should be confirmed in controlled studies with representative materials, and the patients should be followed over a long period of time. It seems that stimulants are not an appropriate treatment for subjects with psychopathic personality and antisocial behaviour. PMID- 9857808 TI - [The significance of associated malformations of the central nervous system in myelomeningocele]. AB - Neural tube defects are the most frequent congenital structural malformations in Norway. Approximately half of these are myelomeningocele. Infants with myelomeningocele frequently have hydrocephalus at birth. Problems with intracranial pressure may as well develop later. There are several reports on pathological corpus callosum and an increasing number of reports on cognitive problems in patients with myelomeningocele. Most patients with myelomeningocele have a tethered spinal cord, and some have syringomyelia. Chiari malformation type II is a malformation of the skull and brainstem which is frequently observed in individuals with myelomeningocele. Chiari malformation may cause severe respiratory problems in infants. Chiari malformation, tethered cord as well as syringomyelia are associated with a range of neurological problems which may progress in adulthood. Surgical intervention may improve the situation. Anaesthesia may induce neurological complications in individuals with Chiari malformation or syringomyelia. Since pregnancy and childbirth are associated with complications, women with myelomeningocele should be examined before they become pregnant. Children and adults with myelomeningocele should routinely undergo MRI examinations of caput and the spinal cord to clarify their anatomical situation. PMID- 9857809 TI - [New distribution forms for pharmaceuticals--a logistic perspective]. AB - Pharmaceuticals are an important input in health care. As a complement to other modes of treatment and as a substitute for hospitalisation, they affect the health of individuals and populations. Enormous public financial resources are spent on pharmaceuticals, and halting the growth in expenditures is a political objective. Factors with room for improvement include drug use efficiency, cost efficient prescription, purchasing prices and distribution. High distribution costs affect prices and, thus, the assessment of product cost vs. utility. Changes in the distribution system may be important, for three reasons: First, increased capital costs call for higher efficiency. Second, increased competition requires improved logistics. And third, information technology has opened up for new supply chain solutions. Direct sales solutions are being considered, and were discussed in a Norwegian public report on the matter, but no final conclusion has been reached. This article discusses changes in the supply of pharmaceuticals and the development of the market. Alternative supply chains are outlined, including what role the postal service may play in a deregulated pharmaceutical market. PMID- 9857811 TI - [Use of the quality of life concept in medical research--useful or useless?]. PMID- 9857810 TI - [Female circumcision--the way forward]. PMID- 9857812 TI - [Estrogen should not be used in primary prevention of cardiovascular disease]. PMID- 9857813 TI - ["The great idea" of Tony Blair]. PMID- 9857814 TI - [Pain in newborn infants]. PMID- 9857815 TI - [Physician on vacation]. PMID- 9857816 TI - [Gene technology can not be prohibited]. PMID- 9857817 TI - [Family practitioner for 1500 patients with 37.5 hours of work weekly?]. PMID- 9857818 TI - Global AIDS surveillance. Part II. PMID- 9857819 TI - Multiresistant Salmonella typhimurium DT 104, Czech Republic. PMID- 9857820 TI - [Psychological aspects of environmental risks]. PMID- 9857821 TI - [Cognition and emotion in the evaluation of accidents: modelling and empirical studies]. AB - Lay people's evaluation of environmental risks is often based on mass media descriptions of single disastrous events. This article analyzes the cognitive and emotional reactions towards those reports. We assume that characteristic features of the events determine the cognitive evaluation and the rise of the emotions anger and sadness. An intuitive, spontaneous, and schema-based evaluation process is postulated that entails a coherent perception and appraisal of an event. The theoretical assumptions are formally specified and implemented in the computer model ITERA (Intuitive Thinking in Environmental Risk Appraisal). In three experimental studies, we presented manipulated descriptions about negative environmental events. The effects of three experimental variables (knowledge about the riskiness of an action, higher goal of the actor, voluntarity of the actor) were compared with the model's predictions. For anger we found a good correspondence between the empirical results and the model's data. The predicted coherence effects of the cognitive appraisal were confirmed to a great extent. Results for sadness did not conform with model predictions. PMID- 9857822 TI - [Subjective causal scenarios for global environmental change]. AB - Two studies are presented that investigate the assumptions that risk evaluation is based on subjective causal scenarios, and that the cognitive representation of global environmental risks is structured according to five causal levels: human attitudes, human activities, emissions or pollutions, environmental changes, and negative consequences. In study 1, 30 subjects listed in free-response format causes, consequences, and remedial measures for 14 environmental risks. Differences between predictive and diagnostic inferences were found: whereas subjects tend to assign immediate rather than mediated causes, they predominantly assign negative consequences for humans, irrespective of the length of the causal chain that leads to these consequences. In study 2, 41 subjects judged the overall similarity between 25 environmental risks. A multidimensional scaling analysis of these similarity judgments replicates the theoretically assumed five causal levels. Results of both studies support the assumptions that risk evaluation is based on implicit causal hypotheses and that the proposed five level structure adequately describes the cognitive representation of environmental risks. PMID- 9857823 TI - Understanding and valuing environmental issues: the effects of availability and anchoring on judgment. AB - This paper addresses the effects of availability and anchoring-and-adjustment on people's beliefs and values concerning environmental issues. The first three studies focus on lay people's perceptions of the causes of large scale environmental risks, the second series of three studies deals with how people value environmental goods and how much they are prepared to pay to mitigate environmental risks. In studies 1-3 we investigate the effects of availability and anchoring-and-adjustment on estimating the contribution of various factors to large scale environmental risks. Highly complex risks such as acid rain and global warming tend to be associated with multiple causes, and our results show that estimating the role of these causes is clearly affected by availability and anchoring-and-adjustment. Both have sizeable effects and persist over time. Moreover, corrective procedures only seem to have a limited effect. Availability and anchoring-and-adjustment not only play a role in judging the possible causes of risks; they also play a role in research attempting to assess the public's willingness to pay (WTP) to protect our environment. The outcomes of WTP surveys are often used as a tool to help policy decision making. In the second part of this article we present three studies on this issue. Results provide further evidence of the impact of the two heuristics on the outcomes of WTP research. Implications for research and practice are briefly discussed. PMID- 9857824 TI - [Abstract or concrete: the importance of biotechnological descriptions in its evaluation]. AB - Biotechnology is an environmental issue that can be evaluated with regard to the impact of the technology "as such" or with regard to specific products. We assume that the concreteness of presentations of biotechnology has an influence on evaluations of risks and benefits. We conducted two experimental studies. In Study 1, 99 subjects evaluated the strength and importance of pro and contra arguments that were presented in short texts on biotechnology. In study 2, 141 subjects were given short texts on biotechnology as well as 15 scales measuring evaluations of risks and chances. Datareveal an influence of the factor "concreteness" on evaluations of pro arguments and benefits as well as on evaluations of contra arguments and risks. But results point to an interaction between levels of concreteness of descriptions and their specific contents: in the pharmaceutical domain concrete descriptions are evaluated more positively than abstract descriptions, in the agricultural domain concrete descriptions are evaluated more negatively. PMID- 9857825 TI - Sustainable development and quality of life: expected effects of prospective changes in economic and environmental conditions. AB - In the context of "sustainable development", we studied which attributes are important to people's quality of life (QoL) and which changes in QoL people would expect from future economic and environmental improvements or deteriorations. About 200 adult subjects evaluated the relative importance of 22 different QoL attributes. They subsequently indicated expected changes in those attributes, under three different scenarios in which economic and environmental conditions would either improve or deteriorate. On average, subjects judged the QoL attributes "healthy", "family", "environmental quality", "nature" and "safety" to be most important, while "recognition", "comfort", "status" and "spiritual life" were found least important. The most important QoL attributes as well as "security" were judged as more important by women than by men. Also observed were income and age effects on relative attribute importance. Our (Dutch) subjects expected significant and varied negative QoL changes from an environmental deterioration scenario involving either an improved or a deteriorated economy. In contrast, they had mixed positive-negative QoL expectations about environmental improvement combined with economic deterioration. Subjects high in environmental concern assigned greater weight to "environmental" QoL attributes, and they expected environmental improvement versus deterioration to more strongly affect their QoL-attributes "environmental quality", "nature", "health" and "unity with nature", than did subjects low in environmental concern. We conclude that quality of life can be meaningfully conceived as a multi-attribute value concept, useful for assessing the expected effects of future economic and environmental conditions. Suggestions are given for methodological improvement and for further research. PMID- 9857826 TI - [Cognitive and affective components of the judgement of environmental risks]. AB - The goal of the present study was to clarify the determinants underlying people's subjective appraisal of the riskiness of environmental problems for human beings. It is assumed that judgments of environmental risks are a function of their ratings on the relevant psychological risk dimensions. In a psychometric study, 186 subjects evaluated 30 environmental problems on 13 judgment scales. Regression analysis shows that the essential predictors of risk judgments are the perceived frequency of harm, the perceived amount of harm and the emotions evoked by thinking about the threats arising from environmental problems. The results show that the subjective judgment of both the perceived frequency and amount of harm vary according to the risk source. By contrast, the various emotions evoked by the different risk sources are explained above all by interindividual differences. The regression model does not satisfactorily predict the risk judgments for one class of environmental risks, namely the condition of and changes in the biosphere. This finding points to the conclusion that subjects use more specific evaluation criteria to evaluate this class of risks. PMID- 9857827 TI - [The relationship between the perceived threat of environmental problems and the formation of action intentions]. AB - The results of a study using questionnaires at two measuring points in high schools in North Germany are reported and discussed. This article analyzes the students' reasons and motivation for more or less environmentally friendly behavior. An integrated theoretical concept of motivation for human action formed the theoretical basis. The action genesis process was subdivided into the phases of forming motives, intentions and volition. Following a presentation of this theoretical concept and its definition with regard to competing and compatible theoretical hypotheses, the methodological procedure is described whereby the various motivation structures of the pupils questioned were analyzed. Mixed distribution models were used which allow modelling of complex multivariate relationships (latent class analysis and mixed Rasch models). The results show strong positive relationships among the variables of the motivation, the intention and the volition phases, but with isolated area-specific motivation patterns and moderator effects of certain cognitive variables. PMID- 9857828 TI - [The pattern of variables predicting self-reported environmental behavior]. AB - Many of the changes observed in our environment today may be traced back to human action. In addition to a descriptive elaboration of possible predictor variables, the psychological analysis of environmental behavior is directed towards the identification of key variables and the structural relationships among these and behavior. Variables that are suitable to predict environmental behavior need to be identified. In this study, 215 subjects each were drawn from an urban and a rural sample; 85 further subjects were considered to be highly environmentally engaged. Thus the total sample population for this study included 515 subjects. Scales on acquisition of information, values, locus of control, attribution of responsibility, and environmental threat were administered to all subjects. A modified version of the protection-motivation theory formulated by Gardner and Stern (1996) served as a reference model. Simultaneous regression analysis revealed that scales specifically directed to the domain of environmental behavior are well suited to explain environmental actions, especially in the subset of highly engaged persons (R2 = .58). In contrast to both other groups, the acquisition of environmentally specific information was a strong predictor in this group. On the basis of these regression analyses, it is argued that additional predictors--along with the ones used in this study--must be taken into account in groups that do not display extraordinary engagement in environmental matters. PMID- 9857829 TI - Effect of citric acid supplements to a maize-soya diet on the in vitro availability of minerals, trace elements, and heavy metals. AB - The aim of the present study was to evaluate the effect of increasing supplements of citric acid (CTA) on the in vitro availability of mineral (Ca, Mg), trace elements (Mn, Zn) and heavy metals (Pb, Cd). Phytaterich diets (0.8%) based on maize, soybean meal, corn starch and soybean oil (containing 5.0 g Ca, 1.2 g Mg, 50 mg Mn, 22 mg Zn, 10 mg Pb and 5 mg Cd per kg diet) were supplemented with 0, 1, 2, 3 and 4% of CTA. All diets were treated in a multi-enzyme in vitro system followed by equilibrium dialysis after peptic and pancreatic digestion. Under the conditions investigated, dietary addition of CTA enhanced dialysability of calcium, magnesium, manganese and zinc. Furthermore, a dose-dependent increase of lead and cadmium dialysability due to CTA supplementations was evident. PMID- 9857830 TI - A dose titration study on the effect of virginiamycin on gilt/sow and piglet performance. AB - Virginiamycin (VM), a compound with proven performance-promoting properties in pigs, can also be considered as sow performance enhancer. The present dose titration study investigates the long-term effects of VM on gilt/sow and litter performance. A total of 140 healthy gilts, in 35 sets of 2 + 2 siblings each, were used in this trial; gilts in each set were randomly allocated to the four treatments (35 gilts per each treatment group): VM0 = negative control, virginiamycin 0 mg/kg, VM20 = virginiamycin 20 mg/kg, VM40 = virginiamycin 40 mg/kg and VM60 = virginiamycin 60 mg/kg of feed. VM was added to the gilt/sow feed for a period starting from 6 months of age up to conception of the fourth parity, covering three complete breeding cycles (pregnancy, lactation and weaning to-conception interval). The feed given did not contain any other antibacterial agent or performance enhancer. Piglets were given exactly the same quantity of creep feed from the fifth day onwards; it did not contain any antibacterial agent or performance enhancer. The lactation period was the same in all gilts/sows (21 days). Results indicate that VM supplementation of the feed improved gilt/sow performance by: (i) increasing (P < 0.05) the mean body weight at each farrowing, (ii) decreasing (P < 0.05) the mean weight loss from farrowing to weaning, and (iii) decreasing (P < 0.05) the mean weaning-to-conception intervals. VM supplementation of gilt/sow feed also resulted in improved litter size and litter performance in terms of: (i) a higher (P < 0.05) mean number of piglets born alive at each birth, (ii) a higher (P < 0.05) mean number of piglets weaned, (iii) a higher (P < 0.05) mean piglet body weight at each birth, and (iv) a higher (P < 0.05) mean piglet body weight at each weaning. These beneficial effects of VM were more pronounced in the VM40 group. PMID- 9857831 TI - A dose titration study on the effect of virginiamycin on specific blood parameters and milk quality in the sow. AB - The present study investigates the changes in blood cholesterol and total lipids concentrations, as well as changes in milk fat, protein, lactose and total solids content of sows after the long-term administration of virginiamycin (VM) in the feed. Seventy-two (72) healthy Dalland gilts, in 18 groups of four (2 + 2) siblings were used in total. The gilts of each group of siblings were randomly allocated to one of the following four dietary treatments: VM0 = negative control, virginiamycin 0 mg/kg, VM20 = virginiamycin 20 mg/kg feed, VM40 = virginiamycin 40 mg/kg feed and VM60 = virginiamycin 60 mg/kg of feed. Treatments started at the age of 6 months covering three complete breeding cycles (up to the third weaning). The first eight groups of siblings (eight gilts per treatment/32 gilts in total) were used for blood sampling (blood group), while the remaining 10 groups of siblings (10 gilts per treatment/40 gilts in total) were used for milk sampling (milk group). Feeds given at insemination, pregnancy and lactation did not contain any other antibacterial or performance enhancer. The results indicate that all three levels of VM supplementation of gilt/sow feed influenced certain blood parameters by increasing (P < 0.05) both cholesterol and total lipids concentrations on the 30th and 60th day of each pregnancy, at each farrowing and at each weaning. The effect of dietary VM on these blood parameters remained constant throughout three consecutive parities. Moreover, the results indicate a beneficial effect of dietary VM on the milk quality of the sows during the third lactation by increasing (P < 0.05) (i) the mean fat content, (ii) the mean protein content, (iii) the mean lactose content and (iv) the mean total solids content. The highest (P < 0.05) levels in both blood and milk parameters were noticed at the inclusion level of 40 mg/kg of feed. PMID- 9857832 TI - Nephelometric determination of haptoglobin plasma concentrations in fattening pigs. AB - The haptoglobin plasma concentrations in 110 fattening pigs living on three commercial farms and in 28 animals on the university pilot farm were measured using a nephelometric detection method based on an immunoassay. Following calibration of the automated analyser (Nephelometer BN 100) with a human haptoglobin standard, the measurements were performed using anti-human antiserum from rabbits. Differences in age, gender or breed of apparently healthy animals seem to have no influence on the plasma concentration of the protein. The average plasma level of haptoglobin in animals suffering from acute respiratory diseases was significantly higher. Furthermore, an increase was observed in animals without clinical symptoms living on a farm characterized by obvious defects in housing conditions. Comparison studies of the nephelometric method by use of a human or porcine standard for calibration and different anti-human or anti porcine antisera with radial immunodiffusion as a reference method resulted in high correlation coefficients for all variations. Optimal accuracy was obtained by calibrating the analyser with a porcine standard. Haptoglobin determinations have been shown to be a useful tool for health monitoring during the integrated pig-production process, allowing recognition of performance-reducing conditions. The immunonephelometric determination method is suitable for quantifying porcine plasma haptoglobin for routine checks. The use of animal-specific standards for calibration improves the accuracy of this method. PMID- 9857833 TI - Selenite and selenium yeast as feed supplements to growing fattening pigs. AB - Twenty-four cross-bred fattening pigs weighing, on average, 24 kg were divided into three equal groups and fed a basic diet containing 0.1 mg selenium/kg. The diet of group A was supplemented with 0.3 mg selenium/kg derived from selenium yeast, group B with 0.1 mg selenium/kg from the same source, and group C with 0.3 mg/kg from sodium selenite. The pigs were slaughtered after 103 days. There was no significant difference between the mean whole blood selenium concentration of the pigs supplemented with selenium yeast neither after five nor after nine weeks of supplementation (group A, 201 and 213 micrograms/litre; group B 192 and 201 micrograms/litre, respectively), but the group supplemented with selenite had significantly lower concentrations both after five and nine weeks (158 and 159 micrograms/litre, respectively). There were no significant differences between the mean activity of glutathione peroxidase (GSH-Px; in the whole blood of the three groups (group A, 348 and 327 mukat*/litre, group B, 342 and 302 mukat/litre; group C, 332 and 300 mukat/litre, after five and nine weeks, respectively). The concentration of selenium in the liver of the pigs supplemented with organic selenium was significantly higher than in the pigs supplemented with selenite (group A, 0.54 mg/kg wet weight; group B, 0.54 mg/kg; group C, 0.40 mg/kg). No deposits of lipofuscin or ceroid pigments were observed in the histological preparations of liver and heart from any of the pigs. PMID- 9857834 TI - Prolonged sevoflurane, isoflurane and halothane anaesthesia in oxygen using rebreathing or non-rebreathing system in cats. AB - Effects of prolonged sevoflurane, isoflurane and halothane anaesthesia in oxygen on clinical, cardiopulmonary, haematologic, and serum biochemical findings were compared in healthy, premedicated cats breathing spontaneously during 6 h of anaesthesia using rebreathing (semi-closed circuit) or non-rebreathing (Bain coaxial circuit) system. Recovery from anaesthesia with sevoflurane was more rapid than that with halothane or isoflurane in both systems. Respiration and heart rates during sevoflurane anaesthesia were similar to those during isoflurane rather than halothane anaesthesia in both systems. The degree of respiratory acidosis during prolonged sevoflurane anaesthesia was similar to that during isoflurane anaesthesia, and was less than that during halothane anaesthesia in both rebreathing and non-rebreathing systems. Prolonged sevoflurane anaesthesia induced mean arterial pressure similar to isoflurane or halothane anaesthesia in the non-rebreathing system, but it depressed mean arterial pressure less than isoflurane or halothane anaesthesia in the rebreathing system. Time related increase in the arterial carbon dioxide partial pressure was observed during halothane anaesthesia especially in the rebreathing system, however, no significant time-related changes in cardiopulmonary variables were observed during either sevoflurane or isoflurane anaesthesia in both systems. There were no significant differences among sevoflurane, isoflurane and halothane anaesthesia in serum biochemical values in both systems. PMID- 9857835 TI - A method for endoscopic embryo collection and transfer in the rabbit. AB - Thirty-two rabbits were used for endoscopic embryo collection and transfer. For embryo collection midventral laparoscopy and transcervical endoscopy were combined for orthograd flushing of the oviducts and uterine horns. Transfer was performed transcervically under laparoscopic control. The mean number of corpora lutea counted in nine donors was 13.3 +/- 8.4. A total of 72 morulae/blastocysts were obtained. No embryos were recovered when only the uterine horns were flushed. 291 embryos were transcervically transferred to 23 recipients which resulted in 10 pregnant animals at day 12. Two of three slaughtered recipients showed together 4 implantation sites. Four animals delivered 12 pups. PMID- 9857836 TI - Gender influences on performance, mood and recovery sleep in fatigued aviators. AB - Female aviators now are able to serve in combat roles, but few studies have addressed potential differences between the ability of males and females to withstand combat stressors. This study examined responses of men and women to one operational stressor, sleep deprivation. Pilots were tested on flight performance and mood during 40-h periods of sustained wakefulness. Baseline and recovery sleep also were examined. Gender produced no operationally-significant effects of flight performance or recovery sleep. Although mood tests showed that women felt less tense and more energetic than men, there were no interactions between sleep deprivation and gender on either flight performance or psychological mood. PMID- 9857837 TI - Using leg muscles as shock absorbers: theoretical predictions and experimental results of drop landing performance. AB - The use of muscles as power dissipators is investigated in this study, both from the modellistic and the experimental points of view. Theoretical predictions of the drop landing manoeuvre for a range of initial conditions have been obtained by accounting for the mechanical characteristics of knee extensor muscles, the limb geometry and assuming maximum neural activation. Resulting dynamics have been represented in the phase plane (vertical displacement versus speed) to better classify the damping performance. Predictions of safe landing in sedentary subjects were associated to dropping from a maximum (feet) height of 1.6-2.0 m (about 11 m on the moon). Athletes can extend up to 2.6-3.0 m, while for obese males (m = 100 kg, standard stature) the limit should reduce to 0.9-1.3 m. These results have been calculated by including in the model the estimated stiffness of the 'global elastic elements' acting below the squat position. Experimental landings from a height of 0.4, 0.7, 1.1 m (sedentary males (SM) and male (AM) and female (AF) athletes from the alpine ski national team) showed dynamics similar to the model predictions. While the peak power (for a drop height of about 0.7 m) was similar in SM and AF (AM shows a +40% increase, about 33 W/kg), AF stopped the downward movement after a time interval (0.219 +/- 0.030 s) from touch-down 20% significantly shorter than SM. Landing strategy and the effect of anatomical constraints are discussed in the paper. PMID- 9857838 TI - Comparison of the effects of backpack loading and of walking speed on foot-floor contact patterns. AB - Load-bearing over variable distances at diverse speeds constitutes a major component of manual materials handling activities that persist as ergonomic concerns even in the most technologically advanced societies. The mechanisms of slipping and tripping relate in part to the way pedal segments approach or leave the walking surface. To demonstrate the kinematic effects of load and speed, as measured underfoot, i.e. at the worker-substrate interface, 45 young adult males traversed a 40 m walkway unburdened, and with 20%, 30%, 40%, 50% and 60% bodyweight backpack loads, under controlled speed conditions. The same subjects also walked unburdened at 0.83 m s-1 to 2.22 m s-1 (i.e. in 1 km h-1 increments between 3 and 8 km h-1). Telemetric footswitch technology was used to obtain temporal patterns of heel, lateral and medial ball and toe of each foot. The findings of this study, in which subjects were shoeless, help to establish 'baseline' descriptions of foot-floor contact patterns, unaffected by interindividual differences due to shoe or boot rigidity. From these, measured or derived data on temporal statistical parameters were obtained which show that all make- and break-contacts of pedal segments occur absolutely and relatively faster as speed increases, whereas forefoot contacts are made faster and broken slower as loads increase. Under loading the earlier phases of stance simulate an increased speed pattern, whereas the later phases of stance simulate a reduced speed pattern. PMID- 9857839 TI - Symmetry of the elbow kinematics during racing wheelchair propulsion. AB - The purpose of this study was to establish whether bilateral symmetry exists during wheelchair propulsion in the elbow movement pattern of trained wheelchair racers. Seven endurance-trained wheelchair racers volunteered to participate in the study. Each subject was recorded by two-dimensional video analysis while pushing on a single-roller wheelchair ergometer at 14 mph (6.58 ms-1) in their own racing wheelchair. The range of elbow flexion, elbow orientation and selected timing parameters including cycle time and time spent in contact with the handrim (propulsion phase) were obtained from both left and right sides. Wilcoxon Matched Pairs Signed-Ranks tests determined the differences between the left and right sagittal plane images of wheelchair propulsion. Furthermore, the Bland-Altman method determined the agreement between the left and right sagittal plane images. The results indicated that the propulsion phase, elbow height and elbow angular displacement characteristics were not significantly different between right and left sides. The right elbow was higher than the left during the recovery phase, but the magnitude of this difference was only 0.03 m (N.S). Factors associated with chair design and the athlete's posture may have contributed to the small differences noted between left and right sides. In conclusion, the results appear to suggest that the assumption of bilateral symmetry of the elbow movement pattern during wheelchair propulsion is valid for the group. However, it is important to note that asymmetries exist in individuals and further research is warranted. PMID- 9857840 TI - Measuring the tilt of the pelvis. AB - A simple and easy-to-use device for measuring forward-backward rotation of the pelvis is introduced. It consists of a small rod, which can be attached to the skin of the sacrum. This 'antenna' follows the movements of the pelvis. The article describes the use of this device and the calibration of this method; to this end the antenna position is related to the 'actual' rotation of the pelvis, defined as the rotation of a set of easily palpable landmarks. Each landmark is found by palpation and its spatial coordinates are measured with a palpator. The results show that pelvic rotation with respect to a reference position can be obtained with an inaccuracy of 10%. With special care this can be reduced to 5%. PMID- 9857841 TI - Evaluation of work-rest schedules with respect to the effects of postural workload in standing work. AB - The influence of four work-rest schedules (60-min shift-15-min break, 45-15, 30 15, 30-30) on acute effects of physical workload in the back and legs due to standing work was investigated in 12 poultry inspectors. Subjective discomfort in the legs and back, and swelling in the distal lower leg were significantly affected, with the 60-15 schedule leading to a higher postural load as compared with the other schedules. No effect on spinal shrinkage was found. It was concluded that the 60-15 schedule should be avoided. An optimal work-rest schedule considering visco-elastic deformation of the spine would probably involve frequent short breaks, whereas longer breaks would seem more effective considering leg swelling. PMID- 9857842 TI - Working in a moving environment. AB - The present paper provides a review of research and theories concerning the question of how and why working in a moving environment may affect performance. It is argued that performance decrements can be expected to occur as a result of general factors or as a result of specific impairments of particular human skills. General effects happen when environmental motion, simulated or real, reduces motivation (due to motion sickness), increases fatigue (due to increased energy requirements), or creates balance problems. Specific effects of moving environments on task performance may only be expected through biomechanical influences on particular skills such as perception (interference with oculomotor control) or motor skills (such as manual tracking). There is no evidence for direct effects of motion on performance in purely cognitive tasks. PMID- 9857843 TI - Slip, trip and fall accidents occurring during the delivery of mail. AB - This study sought to identify causal factors for slip, trip and fall accidents occurring during the delivery of mail. Analysis of in-house data produced information about accident circumstances for 1734 fall cases. The most common initiating events in delivery falls were slips and trips. Slips most often occurred on snow, ice or grass, while trips tended to involve uneven pavements, obstacles and kerbs. Nearly one-fifth of falls occurred on steps, with step falls requiring longer absence from work than falls on the level. Half of all falls occurred during November-February and three-quarters of falls occurred between 7 and 9 a.m. Incidence rates for female employees were 50% higher than for their male colleagues. Accident-independent methods included interviews with safety personnel and managers, discussion groups with delivery employees, and a questionnaire survey of employees and managers. These techniques provided data on risk factors related to the task, behaviour, footwear and equipment. Arising from these accident-independent investigations, it is suggested that unsafe working practices, such as reading addresses while walking and taking shortcuts, increase the risk of falls. Organizational issues include management safety activities, training and equipment provision. Measures are discussed that might lead to a reduction in the incidence of delivery fall accidents. PMID- 9857844 TI - Biological filtering and ecological machinery for self-purification and bioremediation in aquatic ecosystems: towards a holistic view. AB - According to one of the approaches to the definition of criteria for the phenomenon of life, the key attribute is the ability of the system for some self regulating and self-supporting. Part of such holistic functions of aquatic ecosystems as self-regulating and self-supporting is their cleaning the water via a multitude of various mechanisms. The goal of this paper is to present some fundamental elements of the theory of ecosystem self-purification which emphasizes the importance of the four functional biological filters that are instrumental in purification and upgrading the quality of water in aquatic ecosystems. These functional filters are: (1) direct water filtering by aquatic organisms that are filter-feeders; (2) the filter (represented mainly by communities of aquatic plants/periphyton) which prevents input of pollutants and biogenic elements (N, P) from land into water bodies; (3) the filter (represented by benthic organisms) which prevents re-entry of pollutants and biogenic elements from the bottom sediments into the water; (4) the filter (represented by microorganisms attached to particles which are suspended in the water) that provides microbiological treatment of water column. New experimental data by the author reveal the role of man-made effects on the ecological machinery which purifies water. The analysis and discussion lead to the holistic theory of the natural process of bioremediation of aquatic ecosystems. PMID- 9857845 TI - A short history of plant virology. II. The twenties. AB - Plant virology, born at the end of the last century, consolidated during the Twenties. Important new viral diseases were described and their causal agents partially purified and characterized not least because of the development of methods and techniques. An interesting debate concerned the so called "intracellular bodies", which were finally demonstrated to be aggregates of virus particles. Discoveries mainly arrived from experimental investigations on tobacco mosaic virus, and concerned the identification of strains, the demonstration of antigenic property, and the protein nature of viruses. A new concept of virus drew great advantage from the first attempts of classification and nomenclature, and a debate on the living or non-living nature of viruses, universally accepted as a new class of pathogens, was opened. The idea of viruses as self-reproducing particles was first advanced, although on a controversial basis. On the contrary, there was only erratic investigations on the physiological alterations produced by viral diseases in plants and on the relationships between viruses and vectors. In spite of this gaps, the Twenties must be mentioned as the years of the turning point towards a biochemical concept of viruses, which will be achieved in the next decade. PMID- 9857846 TI - The role of synaptic junctions in the identification of human consciousness. AB - Consciousness rests on a complexly regulated mechanism (comprehending the stages of exocytosis and active transport) of projection of sensorial inputs, which can be assembled in quantic sources and then processed along the USC (Unified Synaptic Channel). Such a channel runs in loops all along the labyrinthine structure of the cerebral cortex and constitutes an anatomical-histological structure on its own along which the flow of molecular neurotransmitters determines a sort of constant low-noise effect. PMID- 9857847 TI - Relieving the common cold. PMID- 9857848 TI - I've heard that people with heartburn are at increased risk for developing cancer of the esophagus. Is this true? PMID- 9857850 TI - Hemochromatosis. Iron overload is often overlooked. PMID- 9857849 TI - Nipping colon cancer in the bud. PMID- 9857851 TI - A second look at HRT and heart attack prevention. PMID- 9857852 TI - Beta-blockers and heart attack. PMID- 9857853 TI - Sunlight boosts cataract risk. PMID- 9857854 TI - Effects of partial left ventriculectomy on left ventricular performance in patients with nonischemic dilated cardiomyopathy. AB - OBJECTIVES: This study sought to assess the effects of partial left ventriculectomy (PLV) on left ventricular (LV) performance in a series of consecutive patients with nonischemic dilated cardiomyopathy. BACKGROUND: Reduction of LV systolic function in patients with heart failure is associated with an increase of LV volume and alteration of its shape. Recently, PLV, a novel surgical procedure, was proposed as a treatment option to alter this process in patients with dilated cardiomyopathy. METHODS: We studied 19 patients with severely symptomatic nonischemic dilated cardiomyopathy, before and 13+/-3 days after surgery, and 12 controls. Single-plane left ventriculography with simultaneous measurements of femoral artery pressure was performed during right heart pacing. RESULTS: The LV end-diastolic and end-systolic volume indexes decreased after PLV (from 169 to 102 ml/m2, and from 127 to 60 ml/m2, respectively, p < 0.0001 for both). Despite a decrease in LV mass index (from 162 to 137 g/m2, p < 0.0001), there was a significant decrease in LV circumferential end-systolic and end-diastolic stresses (from 277 to 159 g/cm2, p < 0.0001 and from 79 to 39 g/cm2, p = 0.0014, respectively). Ejection fraction improved (from 24% to 41%, p < 0.0001); the stroke work index remained unchanged. CONCLUSIONS: The PLV improves LV performance by a dramatic reduction of ventricular end systolic and end-diastolic stresses. Further studies are needed to assess whether this effect is sustained during long-term follow-up and to define the role of PLV in the treatment of patients with dilated cardiomyopathy. PMID- 9857855 TI - The current role of left ventricular reduction for treatment of heart failure. PMID- 9857857 TI - The mechanism of decrease in dynamic mitral regurgitation during heart failure treatment: importance of reduction in the regurgitant orifice size. AB - OBJECTIVES: The purpose of this study was to quantify and characterize the regurgitant flow pattern and regurgitant orifice area in patients undergoing therapy for severe heart failure using contemporary echocardiographic techniques. BACKGROUND: Mitral regurgitation may be dynamic in patients with heart failure and ultimately correlate with outcome in a group of patients. METHODS: Fourteen patients with severe heart failure felt to require hemodynamic monitoring for the optimization of medical therapy were enrolled. Two-dimensional and Doppler echocardiograms were performed before and following invasively guided therapy. Hemodynamics and standard echocardiographic dimensions were determined as well as regurgitant volume and regurgitant orifice area derived from color M-mode and Doppler measurements. RESULTS: Invasively guided therapy for heart failure was associated with a reduction in weight, filling pressures of the left and right heart, systemic vascular resistance, and echocardiographic left atrial, left ventricular and mitral annular dimensions. The mitral regurgitant volume decreased from 47+/-27 ml before therapy to 14+/-14 ml after therapy; p < 0.001. While therapy for heart failure markedly attenuated the volume of regurgitation, the pattern of regurgitant flow across the mitral valve was not significantly altered. In contrast, there was no difference in the velocity time integral of the continuous-wave Doppler spectra of mitral regurgitation with therapy (128+/ 23 cm to 123+/-25 cm, p = 0.23). In all patients, the regurgitant orifice area decreased with therapy from 0.55+/-0.38 cm2 to 0.21+/-0.20 cm2 (p < 0.001). CONCLUSIONS: Pharmacologic reduction in filling pressure and systemic vascular resistance leads to a reduction in the dynamic mitral regurgitation of heart failure through a reduction in the regurgitant orifice area but not through a change in the gradient across the mitral valve. Reduction of the regurgitant orifice area is likely related to decreased left ventricular volumes and decreased annular distention. PMID- 9857856 TI - High- versus low-dose ACE inhibition in chronic heart failure: a double-blind, placebo-controlled study of imidapril. AB - OBJECTIVES: To determine dose-related clinical and neurohumoral effects of angiotensin-converting enzyme (ACE) inhibitors in patients with chronic heart failure (CHF), we conducted a double-blind, placebo-controlled, randomized study of three doses (2.5 mg, 5 mg and 10 mg) of the long-acting ACE inhibitor imidapril. BACKGROUND: The ACE inhibitors have become a cornerstone in the treatment of CHF, but whether high doses are more effective than low doses has not been fully elucidated, nor have the mechanisms involved in such a dose related effect. METHODS: In a parallel group comparison, the effects of three doses of imidapril were examined. We studied 244 patients with mild to moderate CHF (New York Heart Association class II-III: +/-80%/20%), who were stable on digoxin and diuretics. Patients were treated for 12 weeks, and the main end points were exercise capacity and plasma neurohormones. RESULTS: At baseline, the four treatment groups were well-matched for demographic variables. Of the 244 patients, 25 dropped out: 3 patients died, and 9 developed progressive CHF (3/182 patients on imidapril vs. 6/62 patients on placebo, p < 0.05). Exercise time increased 45 s in the 10-mg group (p = 0.02 vs. placebo), but it did not significantly change in the 5-mg (+16 s), and 2.5-mg (+11 s) imidapril group, compared to placebo (+3 s). Physical working capacity also increased in a dose related manner. Plasma brain and atrial natriuretic peptide decreased (p < 0.05 for linear trend), while (nor)epinephrine, aldosterone and endothelin were not significantly affected. Renin increased in a dose-related manner, but plasma ACE activity was suppressed similarly (+/-60%) on all three doses. CONCLUSIONS: Already within 3 months after treatment initiation, high-dose ACE inhibition (with imidapril) is superior to low-dose. This is reflected by a more pronounced effect on exercise capacity and some of the neurohormones, but it does not appear to be related to the extent of suppression of plasma ACE. PMID- 9857858 TI - Acute hemodynamic effects of biventricular DDD pacing in patients with end-stage heart failure. AB - OBJECTIVES: The aim of this study was to assess the potential acute benefit of multisite cardiac pacing with optimized atrioventricular synchrony and simultaneous biventricular pacing in patients with drug-refractory congestive heart failure (CHF). BACKGROUND: Prognosis and quality of life in severe CHF are poor. Various nonpharmacological therapies have been evaluated but are restricted in their effectiveness and applications. In the early 1990s, dual chamber pacing (DDD) pacing was proposed as primary treatment of refractory CHF but results were controversial. Recently, tests to evaluate the effect of simultaneous pacing of both ventricles have elicited a significant improvement of cardiac performance. METHODS: Acute hemodynamic study was conducted in 18 patients with severe CHF (New York Heart Association class III and IV) and major intraventricular conduction block (IVCB) (QRS duration = 170+/-37 ms). Using a Swan-Ganz catheter, pulmonary artery pressure, pulmonary capillary wedge pressure (PCWP) and cardiac index (CI) were measured in different pacing configurations: atrial pacing (AAI) mode, used as reference, single-site right ventricular DDD pacing and biventricular pacing with the right ventricular lead placed either at the apex or at the outflow tract. RESULTS: The CI was significantly increased by biventricular pacing in comparison with AAI or right ventricular (RV). DDD pacing (2.7+/-0.7 vs. 2+/-0.5 and 2.4+/-0.6 l/min/m2, p < 0.001). The PCWP also decreased significantly during biventricular pacing, compared with AAI (22+/-8 vs. 27+/-9 mm Hg; p < 0.001). CONCLUSIONS: This acute hemodynamic study demonstrated that biventricular DDD pacing may significantly improve cardiac performance in patients with IVCB and with severe heart failure, in comparison with intrinsic conduction and single-site RV DDD pacing. PMID- 9857859 TI - Expression and distribution of brain natriuretic peptide in human right atria. AB - OBJECTIVES: We investigated expression of brain natriuretic peptide (BNP) as well as atrial natriuretic peptide (ANP) and their genes in human right atria. Their relations with atrial pressure were also examined. BACKGROUND: The BNP plays a roll in electrolyte-fluid homeostasis such as ANP. The tissue level is reported to be elevated in the failing ventricles. However, expression and transmural distribution of BNP in the atria remain unclear. METHODS: Expression of ANP and BNP was immunohistochemically investigated in the right atrial (RA) specimens from 21 patients who had undergone cardiac surgery. The mRNA of specimens were quantitatively measured by Northern blot analysis and also evaluated by in situ hybridization. In addition, plasma levels of ANP and BNP were measured in the patients. RESULTS: The BNP immunoreactivity was diffusely seen in RA tissue of patients with mean RA pressure (mRAP) of 5 mm Hg or more, but it was noted only in the subendocardial half of the atria of those with mRAP less than 5 mm Hg. There was a significant correlation between the incidence of BNP-positive myocytes and mRAP (r = 0.850, p < 0.0001). Conversely, ANP-positive myocytes were found diffusely in all cases. In Northern blot analysis, the mRNAs levels of ANP and BNP in the atrial tissue were positively correlated with the mRAP (ANP, p = 0.775, p < 0.005 and BNP, p = 0.771, p < 0.005). In situ hybridization confirmed these findings. The mRNA levels were significantly correlated to each other (r = 0.845, p < 0.0002). Plasma ANP and BNP levels were elevated in the patients compared with that in controls; however, none were significantly correlated with the mRAP. CONCLUSIONS: Expression of BNP and BNP mRNA is augmented in the atria with increased pressure, and distributed predominantly in the subendocardial side. The level of BNP mRNA was well correlated with that of ANP mRNA. Thus, these two genes might be commonly regulated in response to atrial pressure. PMID- 9857860 TI - Augmentation of the cardiac natriuretic peptides by beta-receptor antagonism: evidence from a population-based study. AB - OBJECTIVES: The present retrospective analysis of data derived from a population based study examined the relationship between intake of beta-receptor antagonists and plasma concentrations of the cardiac natriuretic peptides and their second messenger. BACKGROUND: Beta-receptor antagonists are widely used for treatment of cardiovascular disease. In addition to direct effects on heart rate and cardiac contractility, recent evidence suggests that beta-receptor antagonists may also modulate the cross talk between the sympathetic nervous system and the cardiac natriuretic peptide system. METHODS: Plasma concentrations of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and their second messenger cyclic guanosine monophosphate (cGMP) were assessed in addition to anthropometric, hemodynamic and echocardiographic parameters in a population-based sample (n = 672), of which 80 subjects used beta-receptor antagonists. RESULTS: Compared to subjects without medication, subjects receiving beta-receptor antagonists were characterized by substantially elevated ANP, BNP and cGMP plasma concentrations (plus 32%, 89% and 18%, respectively, p < 0.01 each). Analysis of subgroups revealed that this effect was highly consistent and present even in the absence of hypertension, left atrial enlargement, left ventricular hypertrophy or left ventricular dysfunction. The most prominent increase was observed in a subgroup with increased left ventricular mass index. By multivariate analysis, a statistically significant and independent association between beta-receptor antagonism and ANP, BNP and cGMP concentrations was confirmed. Such an association could not be demonstrated for other antihypertensive agents such as angiotensin-converting enzyme inhibitors or diuretics. CONCLUSIONS: Beta-receptor antagonists appear to augment plasma ANP, BNP and cGMP concentrations. The current observation suggests an important contribution of the cardiac natriuretic peptide system to the therapeutic mechanism of beta-receptor antagonists. PMID- 9857862 TI - Coronary interventions at a crossroads: the bifurcation stenosis. PMID- 9857861 TI - Mechanical debulking versus balloon angioplasty for the treatment of true bifurcation lesions. AB - OBJECTIVES: The purpose of this study was to compare the immediate angiographic and long-term results of debulking versus balloon angioplasty for treatment of true bifurcation lesions. BACKGROUND: Previous studies have shown true bifurcation lesions to be a high risk morphological subset for percutaneous transluminal coronary angioplasty (PTCA). Although atherectomy devices have been used to treat bifurcation lesions, no studies have compared the outcomes of these alternative treatment modalities. METHODS: Between January 1992 and May 1997, we treated 70 consecutive patients with true bifurcation lesions (defined as a greater than 50% stenosis in both the parent vessel and contiguous side branch) with conventional PTCA (n = 30) or debulking (with rotational or directional atherectomy) plus adjunctive PTCA (n = 40). Paired angiograms were analyzed by quantitative angiography, and clinical follow-up was obtained in all patients. RESULTS: Acute procedural success was 73% in the PTCA group and 97% in the debulking group (p = 0.01). Major in-hospital complications occurred in two patients in the PTCA group and one in the debulking group. Treatment with atherectomy plus PTCA resulted in lower postprocedure residual stenoses than PTCA alone (16+/-15% vs. 33+/-17% in the parent vessel, and 6+/-15% vs. 39+/-22% in the side branch; p < 0.001 for both comparisons). At 1 year follow-up, the incidence of target vessel revascularization (TVR) was 53% in the PTCA group as compared with 28% in the debulking group (p = 0.05). Independent predictors of the need for repeat TVR were side branch diameter >2.3 mm, longer lesion lengths, and treatment with PTCA alone. CONCLUSIONS: For the treatment of true bifurcation lesions, atherectomy with adjunctive PTCA is safe, improves acute angiographic results, and decreases target vessel revascularization compared to PTCA alone. The benefits of debulking for bifurcation lesions were especially seen in lesions involving large side branches. PMID- 9857863 TI - Adjunctive stent implantation following directional coronary atherectomy in patients with coronary artery disease. AB - OBJECTIVES: This prospective case-control study evaluated the acute and long-term results of stent implantation preceded by debulking of the plaque by means of directional coronary atherectomy. BACKGROUND: In comparison with balloon angioplasty, intracoronary stenting produces a larger luminal diameter, maintains artery patency and reduces the incidence of restenosis. Optimal stent deployment is a pivotal factor for achieving the best results, but the bulk of the atherosclerotic plaque opposes stent expansion and may limit the success of the procedure. Debulking of the plaque may provide a better milieu for optimal stent deployment. METHODS: Directional coronary atherectomy followed by a single Palmaz Schatz stent implantation was attempted in 100 patients. The successes, complications and angiographic results of the combined procedure were evaluated both acutely and during follow-up. Matched patients undergoing successful Palmaz Schatz stent implantation alone during the same period served as controls. RESULTS: Atherectomy followed by stent implantation was performed in 94 patients with 98 lesions; periprocedural complications were observed in four cases. The stenosis diameter decreased from 76+/-9% at baseline to 30+/-13% after atherectomy (p < 0.0001), and 5+/-9% after stent implantation (p < 0.0001); it increased to 27+/-15% at 6-month angiography (p < 0.0001). During the 14+/-10 months of follow-up, none of the patients died or experienced myocardial infarction, but three patients underwent target lesion revascularization. The patients undergoing stent implantation alone achieved smaller acute gains, tended to have a higher late lumen loss, had a higher restenosis rate (30.5% vs. 6.8%, p < 0.0001) and showed a greater incidence of clinical events during follow-up (p < 0.0001). CONCLUSIONS: Debulking atherosclerotic lesions by means of directional coronary atherectomy before stent implantation is a safe procedure with a high success rate and a low incidence of restenosis at follow-up. PMID- 9857864 TI - Scraping of aortic debris by coronary guiding catheters: a prospective evaluation of 1,000 cases. AB - OBJECTIVES: This study was designed to determine the incidence and to quantitate aortic debris retrieved during placement of guiding catheters in patients undergoing percutaneous interventions. BACKGROUND: Studies have shown that atherosclerotic aortic debris predisposes patients to spontaneous or procedurally related ischemic events. METHODS: In 1,000 consecutive percutaneous interventions, the amount of visible atheromatous material from large-lumen guiding catheters was recorded. Clinical characteristics and in-hospital complications were prospectively collected and associated with debris production. RESULTS: Visible aortic debris (1+ to 3+) occurred more frequently with the Judkins left (JL) catheter, followed by the multipurpose (Multi) catheter compared to any other type of guiding catheter (65%, p = 0.001 and 60%, p = 0.01, respectively). Large debris (2+ and 3+) was observed most frequently with the Multi (odds ratio 3.79, C.I. = 2.32 to 6.21, p = 0.001), JL (odds ratio 2.83, C.I. = 1.98 to 4.05, p = 0.001) and voda left (VL) (odds ratio 2.73, C.I. = 1.51 to 4.95, p = 0.001) catheters. The Judkins right (JR) catheter type was least likely to produce any debris (24%, p = 0.001). A history of unstable angina (p = 0.05) or myocardial infarction (p = 0.003) was associated with a decreased incidence of debris production. The presence of debris was not found to be associated with in-hospital ischemic complications. CONCLUSIONS: Studies have shown that atherosclerosis of the aorta is a potential source of systemic embolism in patients undergoing cardiac catheterization. Our study shows that in more than 50% of percutaneous revascularization procedures, guiding catheter placement is associated with scraping debris from the aorta. Design characteristics of the JL, Multi and VL guiding catheters make them most likely to produce such debris. Meticulous attention to allow the debris to exit the back of the catheter is essential to prevent injecting atheromatous debris into the vascular bed. PMID- 9857865 TI - Diabetes mellitus and the clinical and angiographic outcome after coronary stent placement. AB - OBJECTIVES: The objectives of this study were to analyze the clinical and angiographic outcome of diabetic patients with successful coronary stent placement and to compare these results with those achieved after stenting in nondiabetic patients. BACKGROUND: The outcome of diabetic patients treated with stent placement due to coronary artery disease has not been assessed comprehensively. METHODS: This study analyzes a consecutive series of patients with successful stent placement comprising 715 patients with diabetes and 2,839 patients without diabetes. Clinical one year follow-up and angiographic control at 6 months were part of the protocol. Death, myocardial infarction and target lesion revascularization were considered as adverse events. An automated edge detection system was used for the angiographic assessment. The primary clinical endpoint was event-free survival at one year. The primary angiographic endpoint was restenosis rate at 6 months (> or = 50% diameter stenosis). RESULTS: Event free survival was significantly lower in diabetic than in nondiabetic patients (73.1 vs. 78.5%, p < 0.001). Survival free of myocardial infarction was also significantly reduced in the diabetic group (89.9 vs. 94.4% in nondiabetics, p < 0.001). The incidence of both restenosis (37.5 vs. 28.3%, p < 0.001) and stent vessel occlusion (5.3 vs. 3.4%, p = 0.037) was significantly higher in diabetic patients. Diabetes was identified as an independent risk factor for adverse clinical events and restenosis in multivariate analyses. CONCLUSIONS: Patients with diabetes mellitus have a less favorable clinical outcome at one year after successful stent placement as compared to the nondiabetic patients. The clinical follow-up was characterized by a higher incidence of death, myocardial infarction and reinterventions. Diabetic patients also demonstrated an increased risk for restenosis. PMID- 9857866 TI - Serial volumetric (three-dimensional) intravascular ultrasound analysis of restenosis after directional coronary atherectomy. AB - OBJECTIVES: We report the use of three-dimensional (volumetric) intravascular ultrasound (IVUS) analysis to assess serial changes after directional coronary atherectomy (DCA). BACKGROUND: Recent serial planar IVUS studies have described a decrease in external elastic membrane (EEM) area following catheter-based intervention as an important mechanism of late lumen renarrowing. METHODS: Thirty one patients with de novo native coronary lesions treated with DCA in the Serial Ultrasound Restenosis (SURE) Trial and in Optimal Atherectomy Restenosis Study (OARS) were enrolled in this study. Serial IVUS was performed before and after intervention and at 6 months' follow-up. In a subgroup of 18 patients from the SURE trial, IVUS was also performed at 24 h and at 1 month postintervention. Segments, 20-mm-long (200 image slices), were analyzed using a previously validated three-dimensional, computerized, automated edge-detection algorithm. The EEM, lumen, and plaque+media (P+M = EEM-lumen) volumes were calculated. RESULTS: At follow-up, lumen volume was smaller than at postintervention (159+/ 69 mm3 vs. 179+/-49 mm3, p = 0.0003). From postintervention to follow-up, there was a decrease in EEM volume (377+/-107 to 352+/-125 mm3, p < 0.0001), but no change in P+M volume (p = 0.52). The delta lumen volume correlated strongly with deltaEEM volume (r = 0.842, p < 0.0001), but not with deltaP+M volume. In the 18 patients from the SURE Trial, the decrease in lumen and EEM volumes occurred late, between 1 month and 6 months of follow-up. CONCLUSIONS: Volumetric IVUS analysis demonstrated that late lumen volume loss following DCA was a result of a decrease in EEM volume. This was a late event, occurring between 1 and 6 months' postintervention. PMID- 9857867 TI - Prevalence of sudden cardiac death during competitive sports activities in Minnesota high school athletes. AB - OBJECTIVES: Reliable prevalence data would be useful in assessing the impact of sudden cardiac death in young competitive athletes on the community and designing effective preparticipation screening strategies. BACKGROUND: The frequency with which these catastrophes occur is largely unknown. METHODS: We utilized a circumstance unique to Minnesota in which the precise number of participants and deaths due to cardiovascular disease could be ascertained over a substantial period of time based on a long-standing insurance program for catastrophic injury or death, mandatory for all student athletes engaged in interscholastic sports. RESULTS: Over the 12-year period, 1985/1986 to 1996/1997, inclusive, three sudden deaths due to cardiovascular disease occurred in competitive high school athletes (grades 10-12) during competition or practice. At autopsy, 1 each proved to be due to anomalous origin of the left main coronary artery from the right sinus of Valsalva, congenital aortic valve stenosis (with bicuspid valve) and myocarditis. All three athletes were white and male, 16 or 17 years of age; two competed in cross-country/track and one in basketball. During the study period there were 1,453,280 overall sports participations and 651,695 student athlete participants among the 27 high school sports. The calculated risk for sudden death was 1:500,000 participations and 1:217,400 participants per academic year (or 0.46/100,000, annually). Over a 3-year high school career for a student athlete the estimated risk was 1:72,500. CONCLUSIONS: The risk of sudden cardiac death in a population of high school student athletes was small, in the range of one in 200,000 per year, and was higher in male athletes. The rare occurrence of sudden cardiac death in competitive sports underlines the limitations implicit in structuring productive and cost-effective broad-based preparticipation screening strategies for high school athletes. PMID- 9857868 TI - Atrioventricular node fetal dispersion and His bundle fragmentation of the cardiac conduction system in sudden cardiac death. AB - OBJECTIVES: This study sought to examine the frequency of persistent fetal dispersion of the atrioventricular (AV) node and fragmentation of the atrioventricular bundle (His) bundle in the cardiac conduction system of sudden cardiac death cases and control subjects to establish their importance as the cause of death. BACKGROUND: These are two of the most frequent lesions reported in published reports in the cardiac conduction system in unexplained sudden deaths. METHODS: We have studied the conduction system of 347 hearts: 249 hearts from sudden cardiac death cases and 98 control hearts. The sudden cardiac death cases were divided, according to the pathology found, in three groups: group I: ischemic heart disease, 137 cases; group II: nonischemic heart disease, 48 cases, and group III: unexplained sudden cardiac deaths, 64 cases. The control group (group IV) consisted of patients with unnatural deaths and extracardiac natural deaths. RESULTS: Persistent fetal dispersion of the AV node was observed in 70 cases (20.17%) of all groups with a frequency (40.81%) statistically higher in the control group. Fragmentation of the His bundle was observed in 95 cases (31.77%), and the frequency was statistically higher in the control group, too (47.67%). CONCLUSIONS: Persistent fetal dispersion of the AV node and fragmentation of the His bundle can be a normal variation present during many years in life and must not be considered the anatomic substrate for arrhythmias and sudden death without electrocardiographic abnormalities. PMID- 9857869 TI - Autonomic nervous system activity and the spontaneous initiation of ventricular tachycardia. ESVEM Investigators. Electrophysiologic Study Versus Electrocardiographic Monitoring Trial. AB - OBJECTIVES: We hypothesized that neurohormonal activity contributes to the initiation of sustained ventricular tachycardia (VT) as reflected in indices of heart rate variability (HRV). BACKGROUND: Autonomic nervous system activity participates in experimental arrhythmias but clinical studies have been inconsistent. METHODS: Holter electrocardiograms from 53 patients with VT were analyzed. Heart rate variability indices were determined over 5 and 15 min and 24 h and examined for changes before the onset of VT. Heart rate variability indices in the frequency domain included ultra low frequency power (FP) (ULFP): 0-0.0033 Hz; very low FP (VLFP): 0.0033-0.04 Hz; low FP (LFP): 0.04-0.15 Hz; high FP (HFP): 0.15-0.4 Hz; total power (TP); normalized LFP (LFPn); normalized HFP (HFPn), and the ratio: LFP/HFP. RESULTS: Heart rate variability indices were severely diminished: TP: 12,009+/-11,076 ms2; ULFP: 10,087+/-9,565 ms2; VLFP: 1,416+/-1,571 ms2; LFP: 544+/-620 ms2; HFP: 161+/-176 ms2, and LFP/HFP: 3.68+/ 2.83. Heart rate increased before VT (80.4+/-17.3 to 85.3+/-17.4 bpm, p < 0.001). Several HRV variables declined 30 min before VT compared to 24-h values (VLFP: 5.89+/-17.81%, p = 0.031; LFP: -5.23+/-14.3%, p = 0.003; HFP: -4.35+/-13.7%, p = 0.04). LFPn and the LFP/HFP ratio decreased significantly before the onset of VT (-17.7+/-46.9%, p = 0.035 and -8.24+/-38.8%, p = 0.037, respectively), whereas HFPn increased slightly (4.29+/-29.9%, p = 0.097). CONCLUSIONS: Heart rate rose, whereas LFP, LFPn and LFP/HFP fell before the onset of VT. This pattern of changes could be explained by a rise in sympathetic activity and saturation of the HRV signal resulting in dissociation of the average and rhythmical effects of sympathetic activity. These findings suggest that alterations in autonomic activity contributed to arrhythmogenesis in this group of patients. PMID- 9857870 TI - Long-term outcome of patients with drug-refractory atrial flutter and fibrillation after single- and dual-site right atrial pacing for arrhythmia prevention. AB - OBJECTIVES: An initial crossover study comparing dual- and single-site right atrial pacing was performed followed by a long-term efficacy and safety evaluation of dual-site right atrial pacing in patients with drug-refractory atrial fibrillation (AF). Also examined was the efficacy of two single-site right atrial pacing modes (high right atrium and coronary sinus ostium) and the long term need for cardioversion, antithrombotic and antiarrhythmic drug therapies during dual-site atrial pacing. METHODS: Thirty consecutive patients with drug refractory symptomatic AF and documented primary or drug-induced bradycardia were implanted with a dual chamber rate-responsive pacemaker and two atrial leads. Single-site atrial pacing was performed at the high right atrium or the coronary sinus ostium. Continuous atrial pacing was maintained. RESULTS: Mean arrhythmia free intervals increased from 9+/-10 days in the control period preceding implant to 143+/-110 days (p < 0.0001) in single-site right atrial pacing and 195+/-96 days in dual-site right atrial pacing (p < 0.005 versus single-site pacing and p < 0.0001 versus control). Dual-site right atrial pacing significantly increased the proportion of patients free of AF recurrence (89%) as compared to single-site right atrial pacing (62%, p = 0.02). High right atrial pacing and coronary sinus ostial pacing had similar efficacy for AF prevention. Effective rhythm control was achieved in 86% of patients during dual right atrial pacing. Seventy-eight percent of patients at 1 year and 56% at 3 years remained free of symptomatic AF. The need for cardioversion was reduced after pacemaker implant (p < 0.05) and antithrombotic therapy was reduced (p < 0.06) without any thromboembolic event. Coronary sinus ostial lead dislodgement was not observed after discharge. CONCLUSIONS: Atrial pacing in combination with antiarrhythmic drugs eliminates or markedly reduces recurrent AF. Prevention of AF is enhanced by dual-site right atrial pacing. High right atrial and coronary sinus ostial pacing do not differ in efficacy. Dual-site right atrial pacing is safe, achieves long-term rhythm control in most patients, decreases the need for cardioversion, and antithrombotic therapy can be selectively reduced. PMID- 9857871 TI - Electrical storm in patients with transvenous implantable cardioverter defibrillators: incidence, management and prognostic implications. AB - OBJECTIVES: The purpose of this study was to determine the precise incidence, therapeutic options and prognostic implications of electrical storm in patients with transvenous implantable cardioverter-defibrillator (ICD) systems. BACKGROUND: Approximately 50% to 70% of patients treated with an ICD receive appropriate device-based therapy within the first 2 years. Most arrhythmic events require only one appropriate ICD firing for termination. However, some patients receive multiple appropriate shocks during a short period of time, a condition referred to as "arrhythmic or electrical storm." METHODS: This prospectively designed observational study comprised 136 recipients of transvenous ICDs who were followed for 403+/-242 days. Electrical storm was defined as ventricular tachycardia or fibrillation resulting in device intervention > or = 3 times during a single 24-h period. RESULTS: During follow-up, 57/136 patients (42%) received appropriate ICD therapy. Electrical storm occurred in 14/136 patients (10%) at an average of 133+/-135 days after ICD implantation. The mean number of arrhythmic episodes constituting electrical storm was 17+/-17 (range: 3 to 50; median 8) per patient. In 12 patients, electrical storm required hospital admission. The arrhythmia cluster could be terminated by a combined therapy with beta-blockers and intravenous amiodarone whereas class I antiarrhythmic drugs were only occasionally successful. The cumulative probability of survival as estimated by the Kaplan-Meier method showed that patients with an episode of electrical storm did not have a worse outcome compared to those without such an event. CONCLUSIONS: Electrical storm represents a frequent event in patients treated with modern ICDs. It occurs most commonly late after ICD implantation and can be managed by combined therapy with beta-blockers and amiodarone. Electrical storm does not independently confer increased mortality. PMID- 9857872 TI - Risk factors for clinically important adverse events after protamine administration following cardiopulmonary bypass. AB - OBJECTIVES: The purpose of this study was to determine risk factors for adverse events following protamine administration after cardiopulmonary bypass. BACKGROUND: Intravenous protamine administration is associated with a risk of severe systemic reactions. However, risk factors for these events have not been well delineated, thus hampering development of preventive strategies. METHODS: A case-control study nested within a cohort of consecutive patients undergoing surgery requiring cardiopulmonary bypass was performed. The primary case definition included those events (pulmonary hypertensive and systemic hypotensive) occurring within 10 min of protamine administration in the absence of other measurable causes of hemodynamic compromise. RESULTS: Comparing the 53 cases to the 223 control subjects, three risk factors were independently associated with events (multivariable odds ratio [95% confidence interval]): neutral protamine Hagedorn insulin use (8.18 [2.08, 32.2]); fish allergy (24.5 [1.24, 482.3]), and a history of nonprotamine medication allergy (2.97 [1.25, 7.07]). These risk factors demonstrated an increasingly strong association with progressively more specific case definitions. An estimated 39% of cardiopulmonary bypass patients had one or more of these risk factors. Prior intravenous protamine, central venous pressure prior to protamine, preoperative ejection fraction and the need for inotropes when coming off bypass did not exhibit statistically significant associations with events (all p > 0.15). Prior protamine allergy was associated specifically with an increased risk of pulmonary hypertension (multivariable odds ratio 189; 95% confidence interval 13, 2,856). CONCLUSIONS: Immunologic factors are important in predisposing individuals to protamine reactions, and a substantial proportion of patients are at considerably increased risk Strategies to reduce the risk of protamine-associated events are needed. PMID- 9857873 TI - Restricted coronary flow reserve in patients with mitral regurgitation improves after mitral reconstructive surgery. AB - OBJECTIVES: The purpose of this study was to assess coronary flow characteristics in patients with chronic mitral regurgitation (MR). BACKGROUND: Coronary flow reserve (CFR) has been reported to be restricted in cases with left ventricular (LV) volume overload caused by aortic regurgitation and increased LV preload. METHODS: The study populations consisted of 31 patients with nonrheumatic chronic MR. Eleven with chest pain and normal coronary arteries served as control subjects. Phasic coronary flow velocities were obtained in the proximal segment of the angiographically normal left anterior descending coronary artery at rest and during hyperemia (0.14 mg/kg/min adenosine infusion intravenously) using a 0.014-in. (0.036 cm), 15-MHz Doppler guide wire. Coronary flow reserve was obtained from the ratio of hyperemic/baseline time-averaged peak velocity (APV). Thirteen cases who underwent mitral valve reconstructive surgery were also studied 1 month after surgery. RESULTS: Compared with control subjects, CFR was significantly reduced in cases with MR (2.1+/-0.5 vs. 33+/-0.6, respectively, p < 0.01) because baseline APV was significantly greater (28+/-8 vs. 19+/-6 cm/s, respectively, p < 0.01), although maximal hyperemic APV was not significantly different (56+/-14 vs. 61+/-16 cm/s, respectively, p = NS). Significant correlations were obtained between CFR and LV end-diastolic pressure (LVEDP) (r = 0.70, p < 0.01), LV mass index (r = 0.42, p < 0.01), LV end-diastolic volume (r = 038, p = 0.04) and MR volume (r = 0.39, p = 0.03), and stepwise regression analysis showed LVEDP was the most important determinant of CFR in MR (r2 = 0.49, p < 0.0001). This restricted CFR improved significantly after mitral valve reconstructive surgery (2.1+/-0.5 vs. 3.1+/-0.6, respectively, p < 0.01) because of reduction of baseline APV (28+/-8 vs. 21+/-8 cm/s, respectively, p < 0.01). CONCLUSIONS: Coronary flow reserve is limited in cases with MR because of elevation of baseline resting flow velocity. This reduction of CFR correlates well with increase in LV preload, mass and volume overload, especially with increase in LV preload, and this restricted CFR improves after mitral valve surgery. PMID- 9857874 TI - Variation of anatomic valve area during ejection in patients with valvular aortic stenosis evaluated by two-dimensional echocardiographic planimetry: comparison with traditional Doppler data. AB - OBJECTIVES: Flow variations can affect valve-area calculation in aortic stenosis and lead to inaccuracies in the evaluation of the stenosis. Knowing that transvalvular flow varies normally within one beat, we designed this study to assess the response of the valve to intrabeat variation of flow during systole. Results were compared with flow-derived measurements. BACKGROUND: Technological improvements now allow us to evaluate aortic valve area directly by short axis planimetry. This offers the possibility to perform serial planimetries during one ejection phase and analyze the intrabeat dynamic behavior of the stenotic-aortic valve and compare these measurements with flow-derived measurements. METHODS: Forty echocardiograms displaying different degrees of aortic stenosis were analyzed by frame-by-frame planimetry of the valve area from onset of opening to complete closure. Maximal-mean area, opening and closing rates and ejection times were obtained and compared with Doppler-derived data. RESULTS: Valve area varied during ejection. Stenotic valves opened and closed more slowly than normals and remained maximally open for a shorter period. Mean area by Doppler data corresponded more closely to maximal than to mean-planimetered area. Duration of flow was shorter than valve opening in severely stenotic valves. Discrepancies between Doppler-derived and two-dimensional (2D) measurements decreased in less stenotic valves. CONCLUSIONS: Our observations reveal striking differences between the dynamics of normal and stenotic valves. Surprisingly, Doppler-derived mean-valve area correlated better with maximal-anatomic area than with mean anatomic area in patients with aortic stenosis. Discrepancies between duration of flow and valve opening could explain this phenomenon. PMID- 9857875 TI - Exercise-induced abnormal blood pressure responses are related to subendocardial ischemia in hypertrophic cardiomyopathy. AB - OBJECTIVES: We examined by thallium-201 scintigraphy whether exercise-induced abnormal blood pressure response (BPR) is related to myocardial ischemia. BACKGROUND: Hemodynamic instabilities during exercise in patients with hypertrophic cardiomyopathy (HCM) are considered to be caused by abnormal reflex control of vascular resistance. METHODS: In 105 patients with HCM, exercise thallium scintigraphy was performed by means of a multistage, symptom-limited bicycle ergometer exercise test. RESULTS: Eighty-eight patients had normal BPR (> or = 25 mm Hg from baseline to peak exercise), and 17 had abnormal BPR (<25 mm Hg). Clinical characteristics including age, the prevalence of obstruction, New York Heart Association functional class and echocardiographic measurements were similar between the two groups. Left ventricular end-diastolic pressure was significantly higher in patients with abnormal BPR than in those with normal BPR (18+/-8 vs. 14+/-5 mm Hg, p < 0.05). Exercise-induced perfusion abnormalities including fixed and reversible perfusion defects, and left ventricular cavity dilatation (LVCD) were identified in 72 (69%) of 105 study patients. Left ventricular cavity dilatation indicates subendocardial hypoperfusion and is a marker of diffuse subendocardial ischemia. The prevalence of fixed or reversible perfusion defects was similar between the two groups. Patients with abnormal BPR had the higher prevalence of LVCD as compared to those with normal BPR (47.1 vs. 10%, p < 0.0002). Multiple logistic regression analysis revealed that LVCD was independently associated with abnormal BPR (odds ratio 3.76, 95% confidence interval 1.61 to 8.76). CONCLUSIONS: Exercise-induced abnormal BPRs in patients with HCM are related to subendocardial ischemia during exercise. PMID- 9857876 TI - Long-term outcome and prognostic determinants in children with hypertrophic cardiomyopathy. AB - OBJECTIVES: We sought to determine clinical, angiographic, and echocardiographic predictors of survival in children with isolated hypertrophic cardiomyopathy (HCM) in a large pediatric centre. BACKGROUND: Sudden death is a catastrophic outcome of HCM in childhood but has been difficult to predict. Current therapies might provide for improved outcome if factors identifying high risk can be identified. METHODS: Records of 99 patients diagnosed with HCM from 1958 to 1997 at <18 yr were reviewed for clinical, angiographic (n = 62) and echocardiographic (n = 83) predictors of survival outcome. The effects of clinical characteristics on sudden death (including resuscitated sudden death) were individually tested in Cox's proportionate hazard modeling. RESULTS: Seventy-one subjects were male. Median age at diagnosis was 5.0 yr with a medical follow-up interval of 4.8 yr. Thirty-seven of 97 patients had a family history of HCM. Ambulatory electrocardiograms (ECG) in 78 patients demonstrated supraventricular tachycardia in 16 and ventricular tachycardia in 21. Death or resuscitated sudden death occurred in 18 patients. Sudden death rate was 2.7%/yr after age 8 yr. Cox's proportionate survival modeling revealed increased corrected QT interval (QTc) dispersion on ECG (relative risk [RR] 1.61 per 20 ms increment, p < 0.0003), ventricular tachycardia (VT) on ambulatory ECG (RR 3.75, p < 0.006) and myocardial bridging of the LAD coronary (RR 12.0, p < 0.003) to be associated with reduced time to death or resuscitated sudden death. CONCLUSIONS: Detailed assessment of ECGs, ambulatory ECGs, and coronary angiography can assist in identifying which children with HCM are at risk for sudden death. PMID- 9857877 TI - Growth characteristics of the aortic arch after the Norwood operation. AB - OBJECTIVES: We sought to characterize the growth of the reconstructed aortic arch after the Norwood operation (NO). BACKGROUND: The first stage of surgical palliation of hypoplastic left heart syndrome (HLHS), the NO, includes augmentation of the aortic arch with homograft. Growth characteristics of the reconstructed aortic arch, which is comprised of both native aortic tissue and homograft, have not been characterized. METHODS: Retrospectively, we examined the serial echocardiograms of 50 patients with HLHS who underwent NO to determine the diameter of the reconstructed transverse arch. Measurements were taken immediately after NO and at two other points (1 to 11 years of age). In addition, the autopsy specimens of 10 other patients with HLHS who underwent NO were examined to determine the contribution of native aortic tissue to the overall size of the reconstructed arch at the time of death (12 to 34 months). RESULTS: The diameter of the transverse aorta increased after NO in all subjects. Its rate of growth paralleled that seen in the normal population, though the reconstructed arch had a significantly larger diameter throughout childhood. Examination of autopsy specimens demonstrated a mean increase in circumference of the native aortic tissue of 0.67 cm (p value <0.01), whereas there was no significant change in homograft circumference. CONCLUSIONS: After reconstruction of the aortic arch in HLHS, the diameter of the arch continues to increase throughout childhood, and this increase is due to growth of the native aortic tissue. PMID- 9857878 TI - Myocardial blood flow and coronary flow reserve late after anatomical correction of transposition of the great arteries. AB - OBJECTIVES: Myocardial blood flow (MBF) in children late after arterial switch operation (ASO) was investigated quantitatively by positron emission tomography (PET). BACKGROUND: In children with transposition of the great arteries (TGA), ASO is widely accepted as the management of choice. The long-term patency of coronary arteries after surgical transfer to the neo-aorta, however, remains a concern. METHODS: Twenty-two normally developed, symptom-free children were investigated by PET with nitrogen-13 ammonia at rest and during adenosine vasodilation 10+/-1 years after ASO. A subgroup of 15 children (9+/-1 years; group A) had simple TGA and underwent ASO within 20 days after birth while 7 (13+/-3 years; group B) had complex TGA and underwent ASO and correction of associated anomalies later after birth. Ten young, healthy adults (26+/-6 years) served as the control group. RESULTS: Resting MBF was not different between groups. After correction for the rate-pressure product as an index of cardiac work, younger children of group A had significantly higher MBF at rest compared to healthy adults (102+/-29 vs. 77+/-6 ml/100 g/min; p = 0.012) while flow in group B was not different from the other groups (85+/-22 ml/100 g/min; p = NS). Hyperemic blood flows were significantly lower in both groups after ASO compared to normals (290+/-42 ml/100 g/min for group A, 240+/-28 for group B, 340+/-57 for normals; p < 0.01); thus, coronary flow reserve was significantly lower in both groups after ASO compared to healthy adults (3.0+/-0.6 for group A, 2.9+/-0.6 for group B, 4.6+/-0.9 for normals; p < 0.01). CONCLUSIONS: Blood flow measurements suggest decreased coronary reserve in the absence of ischemic symptoms in children late after arterial switch repair of TGA. The global impairment of stress flow dynamics may indicate altered vasoreactivity; however, the prognostic significance of these findings needs to be determined. PMID- 9857879 TI - Circadian activity of the endogenous fibrinolytic system in stable coronary artery disease: effects of beta-adrenoreceptor blockers and angiotensin converting enzyme inhibitors. AB - OBJECTIVES: To examine circadian changes in the sympathovagal balance, the activity of the renin-angiotensin system and hemostatic variables in patients with stable coronary artery disease, and the effects of beta-adrenoceptor blockade and angiotensin-converting enzyme inhibition. BACKGROUND: Sympathovagal balance and key components of the fibrinolytic system show circadian variability. The effects of beta-adrenergic blocking agents and angiotensin-converting enzyme inhibitors on these autonomic and hemostatic rhythms are not well defined. METHODS: Twenty patients with coronary artery disease underwent 24-h Holter monitoring for heart rate variability and blood sampling (6 hourly for 24 hours) after three consecutive treatment phases, (firstly with placebo, then bisoprolol, and finally quinapril). The effects on sympathovagal balance, hemostatic variables and the renin-angiotensin system activity were measured. RESULTS: The fibrinolytic capacity showed marked circadian variation at the end of the placebo phase (p = 0.002), plasminogen activator inhibitor-1 (PAI-1) activity peaking at 06.00 AM when tissue plasminogen activator (tPA) activity was at its nadir. Sympathovagal balance showed a sharp increase at approximately the same time but plasma renin activity did not rise until later in the day. Inspection of the 24-h profiles suggested that bisoprolol reduced sympathovagal balance and the morning peak of PAI-1 activity and antigen, with a small increase in tPA activity, although these changes were not significant. Quinapril produced a substantial rise in renin (p = 0.01) but did not significantly affect either PAI-1 or tPA. Sympathovagal balance was unaffected by quinapril. CONCLUSIONS: In patients with stable coronary artery disease, angiotensin-converting enzyme inhibition with quinapril does not affect either sympathovagal balance or the endogenous fibrinolytic system. Our data suggest that the sympathoadrenal system may modify fibrinolytic activity, judged by the response to beta-adrenoreceptor blockade with bisoprolol. PMID- 9857880 TI - Therapy with nitroglycerin increases coronary vasoconstriction in response to acetylcholine. AB - OBJECTIVES: The purpose of this study was to investigate whether therapy with nitroglycerin (GTN) would lead to abnormal coronary artery responses to the endothelium-dependent vasodilator acetylcholine. BACKGROUND: Nitroglycerin therapy is associated with specific biochemical changes in the vasculature that may lead to increased vascular sensitivity to vasoconstrictors. METHODS: Patients were randomized to continuous transdermal GTN, 0.6 mg/h (n = 8), or no therapy (n = 7), for 5 days prior to a diagnostic catheterization. Patients had similar risk factors for endothelial dysfunction. Quantitative angiography was performed in the morning to measure the mean luminal diameter of the left anterior descending coronary artery (LAD) in response to intracoronary acetylcholine (peak concentration, 10(-4) mol/liter). The transdermal preparation was removed from the GTN group, and 3 h later experimental procedures were repeated. RESULTS: In the morning, the GTN group experienced greater coronary constriction in response to acetylcholine infusion than those not receiving GTN (-19.6+/-4.2 vs. -3.8+/ 3.0%; p = 0.01). Three hours later, the GTN group continued to display greater constriction to acetylcholine (-24.1+/-5.9%) as compared to the non-GTN group ( 1.8+/-4.8%). When the morning and afternoon responses to acetylcholine were compared, the increase in coronary constriction in the GTN group was greater than the change observed in the non-GTN group (p < 0.05). CONCLUSIONS: This study demonstrates that therapy with GTN causes abnormal coronary vasomotor responses to the endothelium-dependent vasodilator acetylcholine, changes that were persistent for up to 3 hours after GTN discontinuation. This nitrate-associated vasomotor dysfunction has implications with respect to the development of nitrate tolerance and the potential for adverse events during nitrate withdrawal. PMID- 9857881 TI - Prognostic value of pharmacological stress echocardiography in women with chest pain and unknown coronary artery disease. AB - OBJECTIVES: In this study we sought to investigate the prognostic value of pharmacological stress echocardiography in women referred for chest pain, having unknown coronary artery disease. BACKGROUND: The noninvasive identification of a high-risk subgroup among women with chest pain and unknown coronary artery disease is an unresolved task to date. METHODS: A total of 456 women (mean [+/ SD] age 63+/-10 years) underwent pharmacological stress echocardiography with either dipyridamole (n = 305) or dobutamine (n = 151) for evaluation of chest pain and were followed-up for 32+/-19 months. None of them had a previous diagnosis of coronary artery disease. RESULTS: No major complication occurred during stress testing. Five tests (1.1%) were prematurely interrupted because of the appearance of side effects. Echocardiographic positivity was identified in 51 patients. During the follow-up, 23 cardiac events occurred: 3 deaths, 10 infarctions and 10 cases of unstable angina; an additional 21 patients underwent coronary revascularization. At Cox analysis, the echocardiographic evidence of ischemia was found as the only independent predictor of hard cardiac events (death, infarction) (odds ratio [OR] = 27.5; 95% confidence interval [CI] = (6.5 to 115.5; p = 0.0000). When spontaneous cardiac events (death, infarction and unstable angina) were considered as endpoints, the positive echocardiographic result (OR = 23.9; 95% CI = 8.6 to 66.8; p = 0.0000) and family history of coronary artery disease (OR = 3.7; 95% CI = 1.5 to 9.1; p = 0.0037) were independently correlated with prognosis. By using an interactive stepwise procedure, the prognostic value of stress echocardiography was found to be incremental to that provided by clinical variables, both considering hard and spontaneous cardiac events as endpoints. The 3-year survival rate for the negative and the positive population was respectively, 99.5% and 69.5% (p = 0.0000) considering hard cardiac events, 99.2% and 50.6% (p = 0.0000) considering spontaneous cardiac events. CONCLUSIONS: Pharmacological stress echocardiography is safe, highly feasible and effective in risk stratification of women with chest pain and unknown coronary artery disease, also when hard endpoints are considered. Its use can have relevant implications in daily clinical practice for selection of patients needing further investigations. PMID- 9857882 TI - Improvement of exercise capacity by sarpogrelate as a result of augmented collateral circulation in patients with effort angina. AB - OBJECTIVES: The purpose of this study was to evaluate whether a serotonin blocker, sarpogrelate, improves exercise capacity as a result of vasodilation of coronary collateral channels in patients with effort angina. BACKGROUND: Serotonin has been reported to decrease coronary collateral blood flow by collateral vasoconstriction in a canine model, suggesting that platelet activation in feeding coronary arteries of the collateral network has the potential to cause collateral vasoconstriction. METHODS: The subjects consisted of 22 patients with effort angina and reproducible ischemic threshold (group A, 11 patients with thrombolysis in myocardial infarction (TIMI) grade 2 or 3 flow of the ischemia-related coronary artery and Rentrop's collateral index 0 or 1; group B, 11 patients with TIMI grade 0 or 1 flow and Rentrop's collateral index 2 or 3). We repeated the symptom-limited treadmill exercise test using the Balke Ware protocol and exercise tetrofosmin myocardial perfusion scintigraphy with and without pretreatment with 200 mg orally administered sarpogrelate. Each exercise test was performed at 9:00 a.m. on different days. The order of tests with and without sarpogrelate was randomized. RESULTS: In group A, sarpogrelate increased neither exercise time at 0.1 mV ST depression nor double product at 0.1 mV ST depression. In contrast, in group B sarpogrelate increased the exercise duration at 0.1 mV ST depression from 181+/-112 (SD) to 248+/-131 s (p < 0.05) and also increased the double product at 0.1 mV ST depression by 21% (p < 0.01). The severity score using myocardial perfusion scintigraphy at the same workload was significantly (p < 0.01) decreased by 37% in group B, but not in group A (11%), due to the sarpogrelate treatment. CONCLUSIONS: Sarpogrelate augments flow reserve of the collateral circulation and improves exercise capacity in anginal patients with well-developed collaterals. These findings indicate that a serotonin blocker, sarpogrelate, is useful not only as an antiplatelet drugs, but as an antianginal drug. PMID- 9857883 TI - Comparative prognostic value of automatic quantitative analysis versus semiquantitative visual analysis of exercise myocardial perfusion single-photon emission computed tomography. AB - OBJECTIVES: The purpose of this study was to determine the prognostic value of automatic quantitative analysis in exercise dual-isotope myocardial perfusion single-photon emission computed tomography (SPECT) and to compare the prognostic value of quantitative analysis to semiquantitative visual SPECT analysis. BACKGROUND: Extent, severity and reversibility of exercise myocardial perfusion defects have been shown to correlate with prognosis. However, most studies examining the prognostic value of SPECT in chronic coronary artery disease (CAD) have been based on visual analysis by experts. METHODS: We studied 1,043 consecutive patients with known or suspected CAD who underwent rest Tl 201/exercise Tc-99m sestamibi dual-isotope myocardial perfusion SPECT and were followed up for at least 1 year (mean 20.0+/-3.7 months). After censoring 59 patients with early coronary artery bypass grafting or percutaneous transluminal coronary angioplasty, <60 days after nuclear testing, the final population consisted of 984 patients (36% women, mean age 63+/-12 years). RESULTS: During the follow-up period, 28 hard events (14 cardiac deaths, 14 nonfatal myocardial infarctions) occurred. Patients with higher defect extent (>10%), severity (>150) and reversibility (>5%) by quantitative SPECT defect analysis, as well as those with an abnormal scan (>2 abnormal segments, summed stress score >4 and summed difference score >2) by semiquantitative visual SPECT analysis, had a significantly higher hard event rate compared to patients with a normal scan (p < 0.001). With both visual and quantitative analyses, hard event rates of approximately 1% with normal scans and 5% with abnormal scans (p > 0.05) were observed over the 20-month follow-up period. A Cox proportional hazards regression model showed that chi-square increased similarly with the addition of quantitative defect extent and visual summed stress score variables after considering both clinical and exercise variables (improvement chi-square = 11 for both, p < 0.0007). There were no significant differences in the areas under receiver operating characteristic curves between quantitative and visual analysis (p > 0.70). Linear regression analysis also indicated that quantitative assessments correlated well with visual semiquantitative assessments. CONCLUSIONS: The findings of this study indicate that automatic quantitative analysis of exercise stress myocardial perfusion SPECT is similar to semiquantitative expert visual analysis for prognostic stratification. These findings may be of particular clinical importance in laboratories with less experienced visual interpreters. PMID- 9857884 TI - Abciximab in primary coronary angioplasty for acute myocardial infarction improves short- and medium-term outcomes. AB - OBJECTIVES: The purpose of this study was to compare the outcome of primary percutaneous transluminal coronary angioplasty for acute myocardial infarction (MI) when performed with or without the platelet glycoprotein IIb/IIIa antibody, abciximab. BACKGROUND: Abciximab improves the outcome of angioplasty but the effect of abciximab in primary angioplasty has not been investigated. METHODS: Data were collected from a computerized database. Follow-up was by telephone or review of outpatient or hospital readmission records. RESULTS: A total of 182 consecutive patients were included; 103 received abciximab and 79 did not. The procedural success rate was 95% in the two groups. At 30-day follow-up, the composite event rate of unstable angina, reinfarction, target vessel revascularization and death from all causes was 13.5% in the group of patients who did not receive abciximab, 4% (p < 0.05) in the abciximab group and 2.4% (p < 0.05) in the subgroup of patients (n = 87) who completed the 12-h abciximab infusion. At the end of follow-up (mean 7+/-4 months), the composite event rate was 32.4%, 17% (p < 0.05) and 13.1% (p < 0.01) in these three categories respectively. Abciximab bolus followed by a 12-h infusion was an independent predictor of event-free survival, in a Cox proportional hazards model (relative risk 0.49; 95% confidence interval 0.24 to 0.99; p < 0.05). CONCLUSIONS: Abciximab given at the time of primary angioplasty may improve the short- and medium-term outcome of patients with acute MI, especially when a 12-h infusion is completed. PMID- 9857885 TI - Combining thrombolysis with the platelet glycoprotein IIb/IIIa inhibitor lamifiban: results of the Platelet Aggregation Receptor Antagonist Dose Investigation and Reperfusion Gain in Myocardial Infarction (PARADIGM) trial. AB - OBJECTIVES: The trial was designed to assess the safety, pharmacodynamics and effects on reperfusion of the platelet glycoprotein (GP) IIb/IIIa inhibitor lamifiban when given with thrombolysis to patients with ST segment elevation acute myocardial infarction. BACKGROUND: Studies of fibrinolytic agents in acute myocardial infarction have demonstrated a direct relationship between early complete reperfusion and survival. Blockade of the platelet GP IIb/IIIa receptor complex inhibits platelet aggregation and may speed reperfusion when given in conjunction with thrombolysis to patients with acute myocardial infarction. METHODS: Patients with ST segment elevation presenting within 12 h of symptom onset who were treated with either tissue-plasminogen activator or streptokinase were enrolled in this three-part Phase II dose exploration study. In Part A, all patients received the GP IIb/IIIa inhibitor lamifiban in an open-label, dose escalation scheme. Parts B and C were a randomized, double-blind comparison of a bolus plus 24-h infusion of lamifiban versus placebo with patients randomized in a 2:1 ratio. The goal was to identify a dose(s) of lamifiban that provided >85% adenosine diphosphate (ADP)-induced platelet aggregation inhibition. A composite of angiographic, continuous electrocardiographic and clinical markers of reperfusion was the primary efficacy end point, and bleeding was the primary safety end point. RESULTS: Platelet aggregation was inhibited by lamifiban in a dose-dependent manner with the highest doses exceeding 85% ADP-induced platelet aggregation inhibition. There was more bleeding associated with lamifiban (transfusions in 16.1% lamifiban-treated vs. 10.3% placebo-treated patients). Lamifiban induced more rapid reperfusion as measured by all continuous electrocardiographic (ECG) parameters. CONCLUSIONS: Lamifiban given with thrombolytic therapy appears to be associated with more rapid and complete reperfusion than placebo. As expected in this small sample, there were no obvious clinical benefits to lamifiban over placebo. Reconciliation of ECG monitoring with clinical outcomes will require a larger, adequately powered clinical trial. PMID- 9857886 TI - Early changes in myocardial perfusion patterns after myocardial infarction: relation with contractile reserve and functional recovery. AB - OBJECTIVES: The purpose of this study was to assess early temporal changes in myocardial perfusion pattern by myocardial contrast echocardiography (MCE) and their relation to myocardial viability in patients with reperfused acute myocardial infarction (AMI). BACKGROUND: Myocardial contrast echocardiography no reflow is associated with poor contractile recovery after AMI. However, little is known regarding early reversibility of microvascular dysfunction and its relation to myocardial viability. METHODS: Intracoronary MCE was performed immediately after reflow and 9 days later in 28 patients with a first AMI and successful coronary recanalization (Thrombolysis in Myocardial Infarction trial grade 3 flow). Semiquantitative contrast score and wall motion score (WMS) were assessed in each initially asynergic segment at initial and repeat MCE study. Low dose dobutamine echocardiography (DE) was performed at day 10, and follow-up (FU) rest echocardiography was performed 6 weeks later. RESULTS: Among 200 initially asynergic segments, 49% exhibited no or heterogeneous contrast enhancement at initial MCE versus 24% at restudy (p < 0.001). Three groups of segments were defined according to early changes in contrast pattern: group A, "sustained no reflow" (n = 17); group B, improved contrast score (n = 68), and group C, "sustained reflow" (n = 112). Group A segments showed no improvement in WMS at FU. In contrast, group B segments showed significant improvement in WMS at FU (p < 0.0001), and exhibited more frequently contractile reserve at DE (36% vs. 6%, p = 0.02) and contractile recovery at FU (34% vs. 7%, p = 0.03) than group A segments. Group C segments exhibited contractile reserve and contractile recovery in 47% and 51% of segments respectively. CONCLUSIONS: Improvement in MCE perfusion pattern may occur after initial no-reflow in the days following reperfused AMI and is associated with preservation of contractile reserve and gradual regional functional recovery. PMID- 9857887 TI - Clinical characteristics determining the mode of presentation in patients with acute coronary syndromes. AB - OBJECTIVES: The purpose of this study was to examine clinical characteristics of patients with acute coronary syndromes to identify factors that influence the mode of presentation. BACKGROUND: In acute coronary syndromes, presentation with myocardial infarction or unstable angina has major prognostic implications, yet clinical factors affecting the mode of presentation are not well defined. METHODS: A prospective cohort study was made of 1,111 patients with acute coronary syndromes. Baseline demographic, clinical and biochemical data were compared in groups with myocardial infarction (n = 633) and unstable angina (n = 478). RESULTS: The risk of myocardial infarction relative to unstable angina was increased by age >70 years (odds ratio [OR] 2.21; 95% confidence interval [CI] 1.33 to 3.66), male gender (OR 1.56; CI 1.13 to 2.16) and cigarette smoking (OR 1.49; CI 1.09 to 2.03). A rise in admission creatinine from the 10th to the 90th centile of the distribution also increased the odds of myocardial infarction (OR 1.30; CI 1.05 to 1.94). Conversely, the risk of myocardial infarction relative to unstable angina was reduced by previous treatment with aspirin (OR 0.37; CI 0.27 to 0.52), hypertension (OR 0.64; CI 0.47 to 0.86) and previous acute coronary syndromes (OR 0.36; CI 0.26 to 0.51) and revascularization procedures (OR 0.36; CI 0.21 to 0.62). CONCLUSIONS: The clinical presentation of acute coronary syndromes may be influenced by various factors that have the potential to influence the coagulability of the blood, the collateralization of the coronary circulation and myocardial mass. Myocardial infarction is favored by cigarette smoking, advanced age and renal impairment, while unstable angina is favored by treatment with aspirin, hypertension, previous revascularization and previous coronary syndromes. PMID- 9857888 TI - Potential impact of evidence-based medicine in acute coronary syndromes: insights from GUSTO-IIb. Global Use of Strategies to Open Occluded Arteries in Acute Coronary Syndromes trial. AB - OBJECTIVES: The purpose of this study to determine whether use of cardiac medications reflects evidence-based recommendations for patients with non-ST elevation acute coronary syndromes. BACKGROUND: Agency for Health Care Policy and Research practice guidelines for unstable angina recommend the use of cardiac medications based on evidence from randomized trials. It is unknown whether practitioners in the U.S., Canada and Europe follow these recommendations in patients with non-ST elevation acute coronary syndromes. METHODS: We studied 7,743 patients with non-ST elevation acute coronary syndromes enrolled in the international Global Use of Strategies to Open Occluded Arteries in Acute Coronary Syndromes trial. The use of aspirin, beta-adrenergic blocking agents, angiotensin-converting enzyme inhibitors and calcium channel blocking agents was determined at discharge for all patients and "ideal" patients (those with indications and no contraindications). Using published estimates of relative mortality reductions with these drugs, we calculated the lives that could have been saved at 1 year if discharge medication use had better matched guideline recommendations. RESULTS: Overall, guideline adherence at discharge in "ideal" patients was 85.6% for aspirin, 59.1% for beta-blockers and 51.7% for angiotensin converting enzyme inhibitors. Calcium channel blockers were given to 26.7% of patients with a contraindication to these drugs. These rates were similar across locations of enrollment. Women and older patients less often received aspirin when "ideal," and younger patients more often received calcium channel blockers when they were contraindicated. If medication use had been more evidence-based, 1 year mortality might have been reduced by a relative 22%. CONCLUSIONS: There is significant room for improvement in the use of recommended drugs in patients with non-ST elevation acute coronary syndromes. Medication use that more closely follows recommendations could reduce mortality in this population. PMID- 9857889 TI - Enhanced activity of sodium-lithium countertransport in patients with cardiac syndrome X: a potential link between cardiac and metabolic syndrome X. AB - OBJECTIVES: This study was aimed at assessing both stimulated insulinemia and the sodium-lithium countertransport in a selected group of patients with cardiac syndrome X. BACKGROUND: Hyperinsulinemia, which is frequently present in patients with cardiac syndrome X, is often associated with an enhanced activity of the sodium-lithium countertransport, an in vitro marker of sodium-hydrogen exchange. METHODS: Fifteen patients with syndrome X and 14 matched controls were studied. After pharmacological washout, sodium-lithium countertransport was assessed from lithium-loaded red blood cells. Postload insulin levels were evaluated by a double-antibody radioimmunoassay. RESULTS: Maximal velocity of sodium-lithium countertransport was higher in patients with syndrome X compared to controls (635+/-200 vs. 324+/-49 micromol/liter/h, p = 0.001). Fourteen of the 15 patients with syndrome X (93%) presented sodium-lithium countertransport values higher than the mean +2 SD of the control group. At 120 min, 12 patients with syndrome X (80%) had plasma levels of insulin >420 pmol/liter, which corresponds to the mean value +2 SD of controls (p = 0.006). CONCLUSIONS: Both enhanced activity of the sodium-lithium countertransport and stimulated hyperinsulinemia are present in the vast majority of patients with cardiac syndrome X. As enhanced activity of the sodium-lithium countertransport has the potential to cause both glucose intolerance and smooth muscle hyperreactivity, it might represent a common cause of the metabolic and vascular alterations frequently found in syndrome X. PMID- 9857890 TI - Lipoprotein(a) and inflammation in human coronary atheroma: association with the severity of clinical presentation. AB - OBJECTIVES: The purpose of this study was the investigation of the in vivo role of lipoprotein(a) [Lp(a)] and inflammatory infiltrates in the human coronary atherosclerotic plaque and their correlation with the clinical syndrome of presentation. BACKGROUND: Lipoprotein(a) is an atherogenic and thrombogenic lipoprotein, and has been implicated in the pathogenesis of acute coronary syndromes. Lipoprotein(a) induces monocyte chemoattraction and smooth muscle cell activation in vitro. Macrophage infiltration is considered one of the mechanisms of plaque rupture. METHODS: This study of atherectomy specimens investigated the in vivo role of Lp(a) at different stages of the atherogenic process, and its relationship with macrophage infiltration. We examined coronary atheroma removed from 72 patients with stable or unstable angina. Specimens were stained with antibodies specific for Lp(a), macrophages (KP-1), and smooth muscle cells (alpha actin). Morphometric analysis was used to quantify the plaque areas occupied by each of the three antigens, and their colocalization. RESULTS: All specimens had localized Lp(a) staining; the mean fractional area was 58.2%. Ninety percent of the macrophage areas colocalized with Lp(a) positive areas, whereas 31.3% of the smooth muscle cell areas colocalized with Lp(a) positive areas. Patients with unstable angina (n = 46) had specimens with larger mean plaque Lp(a) areas than specimens from stable angina patients (n = 26): 64.4% versus 47.7% (p = 0.004). Unstable angina patients with rest pain (n = 28) had greater mean plaque Lp(a) area than unstable angina patients with crescendo exertional pain (n = 18): 71.1% versus 52.4% (p < 0.001). Mean KP-1 area was 31.2% in unstable rest angina versus 18.3% in stable angina (p = 0.05); alpha-actin area was greater in stable (48.5%) and crescendo exertional angina (48.8%) than in rest angina (30.4%). The strongest correlation between plaque KP-1 and Lp(a) area was in unstable rest angina (r = 0.88, p < 0.001), and between alpha-actin and Lp(a) areas in the crescendo exertional angina (r = 0.62, p < 0.01). CONCLUSIONS: Lipoprotein(a) is ubiquitous in human coronary atheroma. It is detected in larger amounts in tissue from culprit lesions in patients with unstable compared to stable syndromes, and has significant colocalization with plaque macrophages. A correlation of plaque alpha-actin and Lp(a) area suggests a role of Lp(a) in plaque growth. PMID- 9857891 TI - Comparison of mitral inflow and superior vena cava Doppler velocities in chronic obstructive pulmonary disease and constrictive pericarditis. AB - OBJECTIVE: This study was conducted to determine whether Doppler recording of superior vena cava flow velocities can differentiate chronic obstructive pulmonary disease from constrictive pericarditis in patients with a respiratory variation of > or = 25% in mitral inflow E velocity. BACKGROUND: Although respiratory variation (> or = 25%) in mitral E velocity is the main diagnostic criterion for constrictive pericarditis by Doppler echocardiography, it can also be present in chronic obstructive pulmonary disease. Because the respiratory variation is due to increased change in intrathoracic pressure with respiration in chronic obstructive pulmonary disease, and to dissociation of intrathoracic intracardiac pressure changes in constriction, it was hypothesized that the Doppler flow velocity pattern in the superior vena cava (affected by intrathoracic pressure) would be different in these two conditions. METHODS: Pulsed-wave Doppler recording of mitral and superior vena cava flow velocities in 20 patients with chronic obstructive pulmonary disease who had > or = 25% respiratory variation in mitral E-wave velocity were compared with those of 20 patients who had surgically proved constrictive pericarditis. RESULTS: Constrictive pericarditis and chronic obstructive pulmonary disease had similar respiratory variation in mitral E velocity (41% versus 46%). In the latter, the E/A ratio was lower (inspiration, 0.8+/-0.3 versus 1.5+/-0.7 [p < 0.0001]; expiration, 1.0+/-0.3 vs. 1.9+/-0.7 [p < 0.0001]) and deceleration time longer (inspiration, 198+/-53 ms versus 137+/-32 ms; expiration, 225+/-43 ms vs. 161+/ 33 ms [p < 0.0001]). Inspiratory superior vena cava systolic forward flow velocity was significantly higher in chronic obstructive pulmonary disease (72.9+/-22.6 cm/s versus 36.2+/-9.3 cm/s, p < 0.0001), while expiratory systolic forward flow velocity was similar. Hence, there was a significantly greater respiratory variation in superior vena cava systolic forward flow velocity in chronic obstructive pulmonary disease without an overlap with constrictive pericarditis (39.5+/-18.8 cm/s vs. 4.2+/-3.4 cm/s, p < 0.0001). CONCLUSIONS: Despite a similar respiratory variation in mitral E wave velocities, mitral inflow variables in chronic obstructive pulmonary disease are less restrictive compared with those in constrictive pericarditis. More importantly, patients with chronic obstructive pulmonary disease show a marked increase in inspiratory superior vena cava systolic forward flow velocity, which is not seen in patients with constrictive pericarditis. PMID- 9857892 TI - New real-time interactive cardiac magnetic resonance imaging system complements echocardiography. AB - OBJECTIVES: We conducted an initial clinical trial of a newly developed cardiac magnetic resonance imaging (CMRI) system. We evaluated left ventricular (LV) function in 85 patients to compare the clinical utility of the CMRI system with echocardiography, the current noninvasive gold standard. BACKGROUND: Conventional CMRI systems require cardiac-gating and respiratory compensation to synthesize a single image from data acquired over multiple cardiac cycles. In contrast, the new CMRI system allows continuous real-time dynamic acquisition and display of any scan plane at 16 images/s without the need for cardiac gating or breath holding. METHODS: A conventional 1.5T Signa MRI Scanner (GE, Milwaukee, Wisconsin) was modified by the addition of an interactive workstation and a bus adapter. The new CMRI system underwent clinical trial by testing its ability to evaluate global and regional LV function. The first group (A) consisted of 31 patients with acceptable echocardiography image quality. The second group (B) consisted of 31 patients with suboptimal echocardiography image quality. The third group (C) consisted of 29 patients with severe lung disease or congenital cardiac malformation who frequently have suboptimal echo study. Two independent observers scored wall motion and image quality using the standard 16-segment model and rank-order analysis. RESULTS: CMRI evaluation was complete in less than 15 min. In group A, no significant difference was found between ECHO and CMRI studies (p = NS). In group B, adequate visualization of wall segments was obtained 38% of the time using ECHO and 97% of the time using CMRI (p < 0.0001). When grouped into coronary segments, adequate visualization of at least one segment occurred in 18 of 30 patients (60%) with ECHO and in all 30 patients (100%) with CMRI (p < 0.0001). In group C, adequate visualization of the wall segments was obtained in 58% (CI 0.53-0.62) of the time using echocardiography and 99.7% (CI 0.99-1.0) of the time using CMRI (p < 0.0001). CONCLUSIONS: The new CMRI system provides clinically reliable evaluation of LV function and complements suboptimal echocardiography. In comparison with the conventional CMRI, the new CMRI system significantly reduces scan time, patient discomfort and associated cost. PMID- 9857893 TI - HMG-CoA reductase inhibition by atorvastatin reduces neointimal inflammation in a rabbit model of atherosclerosis. AB - OBJECTIVES: To study the effect of the 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA)-reductase inhibitor atorvastatin on the potential mechanisms involved in the recruitment of monocytic cells into the vessel wall. BACKGROUND: Inhibitors of HMG-CoA-reductase reduce cardiovascular mortality though the mechanisms yet elucidated. Most ischemic events are secondary to disruption of atherosclerotic plaques highly infiltrated by macrophages. METHODS: Atherosclerosis was induced in the femoral arteries of rabbits by endothelial damage and atherogenic diet for 4 weeks. Then, animals were switched to standard chow and randomized to receive either no treatment or atorvastatin (5 mg/kg/d) and killed after 4 weeks. RESULTS: Atorvastatin induced a significant reduction in serum lipids and in lesion size. Arterial macrophage infiltration was abolished by the treatment, and monocyte chemoattractant protein-1 (MCP-1) was significantly diminished in the neointima and in the media. Nuclear factor kappa-B (NF-kappaB) was activated in the 60% of the lesions, both in macrophages and vascular smooth muscle cells (VSMC), of the untreated group while only in 30% of the atorvastatin group. NF kappaB activity was also lower in the uninjured aorta and liver of treated compared with untreated rabbits. In cultured VSMC, MCP-1 expression and NF-kappaB activity induced by tumor necrosis factor alpha were downregulated by atorvastatin. CONCLUSIONS: In a rabbit atherosclerosis model, atorvastatin diminishes the neointimal inflammation, and this could contribute to the stabilization of the atherosclerotic plaque. This may be an additional explanation for the reduction of acute ischemic events in patients treated with statins. PMID- 9857894 TI - Encircling overlapping multipulse shock waveforms for transthoracic defibrillation. AB - OBJECTIVES: This study was performed to determine the efficacy of new encircling overlapping multipulse, multipathway waveforms for transthoracic defibrillation. BACKGROUND: Alternative waveforms for transthoracic defibrillation may improve shock success. METHODS: First, we determined the shock success achieved by three different waveforms at varying energies (18-150 J) in 21 mongrel dogs after short duration ventricular fibrillation. The waveforms tested included the traditional damped sinusoidal waveform, a single pathway biphasic waveform, and a new encircling overlapping multipulse waveform delivered from six electrode pads oriented circumferentially. Second, in 11 swine we compared the efficacy of encircling overlapping multipulse shocks given from six electrode pads and three capacitors versus encircling overlapping shocks given from a device utilizing three electrodes and one capacitor. RESULTS: In the first experiment, the encircling overlapping waveform performed significantly better than biphasic and damped sinusoidal waveforms at lower energies. The shock success rate of the overlapping waveform (six pads) ranged from 67+/-4% (at 18-49 J energy) to 99+/ 3% at > or = 150 J; at comparable energies biphasic waveform shock success ranged from 26+/-5% (p < 0.01 vs. encircling overlapping waveforms) to 99+/-5% (p = NS). Damped sinusoidal waveform shock success ranged from 4+/-1% (p < 0.01 vs. encircling overlapping waveform) to 73+/-9% (p = NS). In the second experiment the three electrode pads, one capacitor encircling waveform achieved shock success rates comparable with the six-pad, three-capacitor waveform; at 18-49 J, success rates were 45+/-15% versus 57+/-12%, respectively (p = NS). At 100 J, success rates for both were 100%. CONCLUSIONS: We conclude that encircling overlapping multipulse multipathway waveforms facilitate transthoracic defibrillation at low energies. These waveforms can be generated from a device that requires only three electrodes and one capacitor. PMID- 9857895 TI - Adventitial vasa vasorum in balloon-injured coronary arteries: visualization and quantitation by a microscopic three-dimensional computed tomography technique. AB - OBJECTIVES: The objective of this study was to examine the quantitative response of the adventitial vasa vasorum to balloon-induced coronary injury. BACKGROUND: Recent attention has focused on the role of vasa vasorum in atherosclerotic and restenotic coronary artery disease. However, the three-dimensional anatomy of these complex vessels is largely unknown, especially after angioplasty injury. The purpose of this study was to visualize and quantitate three-dimensional spatial patterns of vasa vasorum in normal and balloon injured porcine coronary arteries. We also studied the spatial growth of vasa vasorum in regions of neointimal formation. A novel imaging technique, microscopic computed tomography, was used for these studies. METHODS: Four pigs were killed 28 d after coronary balloon injury, and four pigs with uninjured coronary arteries served as normal controls. The coronary arteries were injected with a low-viscosity, radiopaque liquid polymer compound. Normal and injured coronary segments were scanned using a microscopic computed tomography technique. Three-dimensional reconstructed maximum intensity projection and voxel gradient shading images were displayed at different angles and voxel threshold values, using image analysis software. For quantitation, seven to 10 cross-sectional images (40 normal and 32 balloon injured cross-sections) were captured from each specimen at a voxel size of 21 microm. RESULTS: Normal vasa vasorum originated from the coronary artery lumen, principally at large branch points. Two different types of vasa were found and classified as first-order or second-order according to location and direction. In balloon-injured coronary arteries, adventitial vasa vasorum density was increased (3.16+/-0.17/mm2 vs. 1.90+/-0.06/mm2, p = 0.0001; respectively), suggesting neovascularization by 28 d after vessel injury. Also, in these injured arteries, the vasa spatial distribution was disrupted compared with normal vessels, with proportionally more second-order vasa vasorum. The diameters of first-order and second-order vasa were smaller in normal compared with balloon-treated coronary arteries (p = 0.012 first-order; p < 0.001, second-order; respectively). The density of newly formed vasa vasorum was proportional to vessel stenosis (r = 0.81, p = 0.0001). Although the total number of vasa was increased after injury, the total vascular area comprised of vasa was significantly reduced in injured vessels compared with normals (3.83+/-0.20% to 5.42+/-0.56%, p = 0.0185). CONCLUSIONS: Adventitial neovascularization occurs after balloon injury. The number of new vessels is proportional to vessel stenosis. These findings may hold substantial implications for the therapy of vascular disease and restenosis. PMID- 9857896 TI - The role of adventitial vasculature in restenosis: a new view of an old problem. PMID- 9857897 TI - "Stunning" of the left atrium after spontaneous conversion of atrial fibrillation to sinus rhythm: demonstration by transesophageal Doppler techniques in a canine model. AB - OBJECTIVES: This study compared left atrial and left atrial appendage contraction velocities in sinus rhythm before and after a brief period of atrial fibrillation in a canine model. BACKGROUND: In patients, left atrial appendage contraction velocities measured during sinus rhythm after cardioversion from atrial fibrillation are depressed relative to left atrial appendage emptying velocities measured during atrial fibrillation, suggesting that the left atrial appendage is mechanically "stunned." METHODS: This phenomenon was studied in a canine model of acute (60 min) pacing-induced atrial fibrillation followed by spontaneous reversion to sinus rhythm using epicardial and transesophageal pulsed wave Doppler. Unique features of the model include: 1) comparison of left atrial function postconversion to baseline sinus rhythm rather than to measurements during atrial fibrillation, 2) control of the duration of atrial fibrillation and 3) elimination of the extraneous influences of direct current shock and antiarrhythmic agents, which may independently depress left atrial function. RESULTS: Hemodynamic conditions (heart rate, mean arterial pressure, cardiac output, mean pulmonary artery pressure, mean right atrial pressure and mean left atrial pressure) at baseline, during 60 min of atrial fibrillation and after reversion to sinus rhythm were constant throughout the study period. Peak left atrial contraction velocities (measured from the transmitral flow velocity profile) were significantly (p < 0.02) reduced to 64+/-22% of baseline values upon spontaneous conversion of atrial fibrillation to sinus rhythm and recovered to basal values by 20 min after resumption of sinus rhythm. Peak left atrial appendage contraction velocities were significantly (p < 0.001) reduced to 49+/ 24% of baseline values upon spontaneous conversion of atrial fibrillation to sinus rhythm and recovered to basal values by 40 min after reversion to sinus rhythm. CONCLUSIONS: Even brief (60 min) periods of atrial fibrillation in normal canine hearts result in marked depression of global left atrial systolic function and regional left atrial (left atrial appendage) systolic function upon resumption of sinus rhythm. This "mechanical stunning" of left atrial systolic function appears to be more profound and of longer duration for the left atrial appendage compared with the left atrium as a whole, which may predispose the appendage to blood stasis and thrombus formation. Chronic models of atrial fibrillation need to be developed to examine the impact of longer periods of atrial fibrillation upon the magnitude and duration of postconversion left atrial "stunning." PMID- 9857898 TI - Artery size, neointima, and remodeling: time for some standards. AB - Controversy continues regarding the mechanism of coronary restenosis. While neointimal thickening was initially considered the major cause, recent studies suggest that changes in arterial size, or remodeling, plays an important or even dominant role in late lumen loss. Moreover, neointimal thickness and remodeling may be interrelated. The field has been complicated by the fact that remodeling analyses have not used consistent definitions or methods. In this editorial we thus describe a quantitative paradigm for remodeling analyses: as arterial plaque or neointima forms in an artery, it is accompanied by luminal encroachment, artery expansion or gradations of either. In this manner, remodeling is generally defined as any arterial size change (enlargement or contraction), independent or dependent of neointimal thickening. Standardization of definitions and quantitative methods may improve understanding of the components of restenosis resulting from artery size changes, neointimal thickening and their impact on lumen size. PMID- 9857899 TI - Therapeutic CPR or consent DNR--a dilemma looking for an answer. PMID- 9857900 TI - What's new in peer review? PMID- 9857901 TI - Heparin for acute myocardial infarction: "news" from the "past". PMID- 9857902 TI - Determinants of Doppler pulmonary flow components. PMID- 9857903 TI - Seasonal variations in the incidence of acute myocardial infarction. PMID- 9857904 TI - Anticoagulation in dilated cardiomyopathy. PMID- 9857905 TI - ATRAMI: a mark in the quest for the prognostic value of autonomic markers. Autonomic Tone and Reflexes After Myocardial Infarction. PMID- 9857906 TI - Coronary collateral perfusion--predictive value of myocardial contrast echocardiography. PMID- 9857908 TI - Stress echocardiography in the community. PMID- 9857907 TI - Surgery for ischaemic mitral regurgitation. PMID- 9857909 TI - The paradoxical benefit of sulfonylurea agents in diabetics undergoing coronary revascularization. PMID- 9857910 TI - Social support and coronary heart disease in women: the challenge to learn more. PMID- 9857911 TI - Growth hormone treatment in chronic heart failure: lessons from the first 99 patients. PMID- 9857912 TI - Platelet glycoprotein IIB/IIIA receptor antagonists. An asset for treatment of unstable coronary syndromes and coronary intervention. PMID- 9857913 TI - The clinical implications of diabetic heart disease. PMID- 9857914 TI - Re-use of devices in cardiology. Proceedings from a Policy Conference at the European Heart House, 5-6 February, 1998. PMID- 9857915 TI - Smoking, lipoproteins and coronary heart disease risk. Data from the Munster Heart Study (PROCAM). AB - AIMS: The mechanism of the increase in coronary heart disease risk associated with smoking is unclear, but may partly be due to smoking-related changes in intermediate risk factors such as lipid levels, fibrinogen and blood pressure. We therefore examined the distribution of these variables among smokers and non smokers in the Munster Heart Study. METHODS: 20696 men, aged 41.7+/-2.7 years (mean +/- SD) and 10212 women, aged 37.0+/-2.6 years, were enrolled between 1978 and 1995. Thirty-two percent of women and 36% of men smoked. Compared to non smokers, mean levels of low density lipoprotein cholesterol, total cholesterol, triglycerides and fibrinogen were increased, respectively, by 1.4%, 0.9%, 15% and 12.1% in male and by 2.0%, 5.5%, 12% and 3.4% in female smokers. Mean high density lipoprotein cholesterol levels, body mass index and blood pressure were reduced, respectively, by 6.4%, 3.8%, and 2% in male, and by 6.7% 1.2% and 2% in female smokers. In the subgroup of 4639 men aged 40 to 65 with 8 years of follow up, the coronary event rate (definite myocardial infarction, sudden cardiac death) in cigarette smokers was more than twice that of non-smokers with otherwise identical risk factors. CONCLUSION: In the Munster Heart Study, smoking was associated with adverse changes in lipids (of greater magnitude in women), and fibrinogen (of greater magnitude in men). However, these changes explained only a small part of the smoking-related increase in coronary heart disease risk. PMID- 9857916 TI - Influence of biological factors on lipid and fibrinogen measurements in young men. An epidemiological study in 2009 recruits. AB - AIMS: The aim of the present study was to detect significant relationships between lipid and fibrinogen measurements and several biological factors in young men. METHODS AND RESULTS: Medical history was obtained, and plasma lipids, lipoprotein (a) and fibrinogen levels were measured in 2009 male Greek army recruits (mean age 22.37+/-3.03 years) not taking any drugs. Plasma levels were as follows: total cholesterol, 171+/-34 mg x dl(-1), low density lipoprotein (LDL) cholesterol, 111+/-34 mg x dl(-1), high density lipoprotein (HDL) cholesterol, 45+/-10 mg x dl(1), and triglycerides, 74+/-32 mg x dl(-1). Lipoprotein (a) and fibrinogen were 18+/-13 and 278+/-67 mg x dl(-1). The atherosclerotic index, calculated as the ratio of total cholesterol/HDL, was 4+/ 1. Analysis of multivariate models that included potentially confounding factors revealed the following: body mass index, season of year during which blood examinations were performed, alcohol consumption, and place of residence were found to be significantly associated with plasma levels of total cholesterol, LDL cholesterol, fibrinogen and the atherosclerotic index in the pooled population. Season and physical activity were significantly associated with HDL-cholesterol, whereas season and family history of acute myocardial infarction were associated with triglycerides levels. Body mass index, family history of myocardial infarction and physical activity were associated with lipoprotein (a). CONCLUSION: Body mass index, season, alcohol consumption and place of residence are markers of plasma lipid profile and fibrinogen in young men. A family history of acute myocardial infarction and physical activity are related to lipoprotein (a). PMID- 9857917 TI - Social relations and extent and severity of coronary artery disease. The Stockholm Female Coronary Risk Study. AB - AIMS: Social relations have been repeatedly linked to coronary heart disease in men, even after careful control for standard risk factors. Women have rarely been studied and results have not been conclusive. We investigated the role of social support in the severity and extent of coronary artery disease in women. METHODS AND RESULTS: One hundred and thirty-one women, aged 30 to 65 years, who were hospitalized for an acute coronary event and were included in the Stockholm Female Coronary Risk Study, were examined with computer assisted quantitative coronary angiography. Angiographic measures included presence of stenosis greater than 50% in at least one coronary artery (severity) and the number of stenoses greater than 20% within the coronary tree (extent). Social factors included two measures of social support, which were previously shown to predict coronary disease in prospective studies of men. After adjustment for age, lack of social support was associated with both measures of coronary artery disease. With further adjustment for smoking, education, menopausal status, hypertension, high density lipoprotein and body mass index, the risk ratio for stenosis greater than 50% in women with poor as compared to those with strong social support was 2.5 (95% confidence interval 1.2 to 5.3; P=0.003). Also, women with poor social support had more stenoses obstructing at least 20% of the coronary lumen with multivariate adjustment, but the difference from women with strong support was only of borderline significance (P=0.09). CONCLUSION: The findings suggest that lack of social support contributes to the severity of coronary artery disease in women, independent of standard risk factors. PMID- 9857918 TI - Angiotensin II type 1 receptor A1166C gene polymorphism. Absence of an association with the risk of coronary artery disease and myocardial infarction and of a synergistic effect with angiotensin-converting enzyme gene polymorphism on the risk of these diseases. AB - AIM: There is evidence that interaction between angiotensin II type 1 receptor A1166C gene polymorphism and angiotensin I-converting enzyme Insertion/Deletion gene variation might have an effect on the risk of myocardial infarction. The study was carried out in a population of 2244 male Caucasians, whose coronary anatomy was defined by means of coronary angiography. We analysed the relationship, on the risk of ischaemic heart disease, of angiotensin II type 1 receptor A1166C gene variation, not only to myocardial infarction but also to coronary artery disease, and its potential interaction with angiotensin I converting enzyme Insertion/Deletion gene polymorphism. METHODS AND RESULTS: No association was detected between angiotensin II type 1 receptor A1166C gene polymorphism and coronary artery disease. Similarly, there was no link to myocardial infarction, either in the total population or in low risk groups. In addition, most importantly, we found no interaction between angiotensin II type 1 receptor A1166C gene variation and angiotensin I-converting Insertion/Deletion polymorphism, either in connection with the risk of coronary artery disease or myocardial infarction. CONCLUSION: This angiotensin II type 1 receptor A1166C gene variation is not associated with any detectable increase in risk of ischaemic heart disease. The findings of the present study do not suggest that, as regards risk of coronary artery disease and myocardial infarction, there is interaction between gene polymorphism and angiotensin I-converting enzyme Insertion/Deletion gene variation. PMID- 9857919 TI - Better increase in fibrin gel porosity by low dose than intermediate dose acetylsalicylic acid. AB - AIM: To investigate the influence on plasma fibrin gel structure of low and intermediate doses of acetylsalicylic acid in healthy individuals. The influence of acetylsalicylic acid on thrombin formation, fibrinolytic capacity and plasminogen inhibitor-1 in plasma was also investigated. METHODS: Nineteen subjects were treated with 75 mg and 11 with 320 mg acetylsalicylic acid daily; eight subjects received both doses. Fibrin gel structure was determined by a permeability technique yielding a porosity constant (Ks), and the thromboxane metabolite 11-dehydro-thromboxane B2 (TxM) was determined by an ELISA. RESULTS: Acetylsalicylic acid increased fibrin porosity by 65% at 75 mg (P<0.001, n=19), whereas lower increases were found at 320 mg (+22%, P<0.05, n=11). One week after withdrawal Ks had essentially returned to baseline (ns). Urinary thromboxane metabolites were suppressed during treatment (-61%, P<0.001 at 75 mg, n=19; -46%, P<0.01 at 320 mg, n=11). The intra-individual comparison showed similar results (Ks +92%, TxM -62% at 75 mg; Ks +5%, TxM -52% at 320 mg). Fibrinolytic capacity, plasminogen inhibitor-1 levels and thrombin generation (in platelet-poor citrated plasma) were not influenced. CONCLUSION: Low dose acetylsalicylic acid causes the greatest increase in fibrin gel porosity; this may well be of therapeutic importance. PMID- 9857920 TI - Risk stratification by pharmacological stress echocardiography in a primary care cardiology centre. Experience in 1082 patients. AB - AIM: In this study we sought to determine the safety, feasibility and prognostic value of pharmacological stress echocardiography performed in a primary care cardiology centre, populated by unselected patients evaluated with the aid of limited financial and technological resources. METHODS AND RESULTS: The study population was 1082 patients undergoing pharmacological stress echocardiography with either dipyridamole (n=714) or dobutamine (n=368) for the evaluation of known or suspected coronary artery disease. The echocardiogram was positive in 284 (26%) patients. Two sustained ventricular tachycardias, reversible by antidote, occurred during stress testing. Limiting ischaemia-independent side effects occurred in 1.5% dipyridamole and in 2.4% dobutamine stress echocardiograms. During follow-up (33+/-18 months), 17 cardiac deaths and 27 non fatal myocardial infarctions occurred. One hundred and twenty-seven patients underwent coronary revascularization, of whom 105 (37%) had a positive and 22 (3%) a negative stress testing result (P<0.0001). At Cox analysis, allowing for 14 clinical and stress-echo variables. the independent predictors of cardiac death were, in decreasing order, a positive stress testing result (Odds ratio [OR]=6.0), resting wall motion score index (OR=5.7), age greater than 65 years (OR=4.9), previous Q-wave myocardial infarction (OR=3.5), and hypercolesterolaemia (OR=2.7). The 4-year survival rate was 99.2% for patients with a negative and 89.8% for patients with a positive stress testing result (P=0.0000). When cardiac hard events (cardiac death and non-fatal myocardial infarction) were considered as end-points, the following variables were independently associated with prognosis: positive result of stress testing (OR=3.1), hypercolesterolaemia (OR=2.4), and resting wall motion score index (OR=2.7). The 4-year infarction-free survival rate was 97.0% for patients with a negative and 81.4% for patients with a positive stress testing result (P=0.0000). CONCLUSIONS: Pharmacological stress echocardiography with either dipyridamole or dobutamine was safe and feasible, providing an excellent tool for prognostic assessment of coronary artery disease in a primary care cardiology centre. PMID- 9857921 TI - Coronary collateral circulation behaviour and myocardial viability in chronic total occlusion treated with coronary angioplasty. AB - AIMS: We explored the role of microcirculation integrity following the chronic occlusion of an infarct-related artery to assess the behaviour of collateral circulation during and after reperfusion by coronary angioplasty METHODS AND RESULTS: Eighteen patients with a proximally occluded left anterior descending artery and firm evidence of intercoronary collateral circulation were studied with selective coronary angiography and selective intracoronary myocardial contrast echocardiography, before coronary angioplasty, and at 5 and 15 min and 12 h later. Myocardial enhancement during myocardial contrast echocardiography was evaluated with a semiquantitative score (0-3), which was correlated to basal and 6 months' regional left ventricular wall motion results. 16/18 procedures were successfully performed; four patients with an inadequate acoustic window were excluded. Restenosis was evident at the 6 months' follow-up in two patients. Basal myocardial contrast echocardiography indicated that 81/192 segments from the left anterior descending coronary artery and 90/192 from the right coronary artery were perfused; no perfusion was observed in 21 segments either before or after coronary angioplasty. After coronary angioplasty, the angiographic intercoronary collateral circulation immediately disappeared, and myocardial contrast echocardiography revealed that there was a progressive reduction of segments perfused by the right coronary artery and an increase in segments perfused by the left anterior descending coronary artery. Regional left ventricular wall motion analysis demonstrated that there was abnormal motion in 51/192 segments. There was no improvement in segments with score 0 and abnormal motion after 6 months (100% sensitivity), but 16/17 segments with score 3 did show an improvement (98% specificity). The predictive value of intermediate scores (1-2) in detecting long-term improvement, was only 43%. CONCLUSION: These data show that the adaptive mechanism observed in the behaviour of epicardial and microvascular circulation after reperfusion of a chronic occluded infarct-related artery can vary. In addition, this study clearly shows that microvascular integrity detected by myocardial contrast echocardiography can provide myocardial viability. PMID- 9857922 TI - Impairment of exercise capacity and peak oxygen consumption in patients with mild left ventricular dysfunction and coronary artery disease. AB - AIMS: Most studies in chronic heart failure have only included patients with marked left ventricular systolic dysfunction (i.e. ejection fraction < or =0.35), and patients with mild left ventricular dysfunction are usually excluded. Further, exercise capacity strongly depends on age, but age-adjustment is usually not applied in these studies. Therefore, this study sought to establish whether (age-adjusted) peak VO2 was impaired in patients with mild left ventricular dysfunction. METHODS: Peak VO2 and ventilatory anaerobic threshold were measured in 56 male patients with mild left ventricular dysfunction (ejection fraction 0.35-0.55; study population) and in 17 male patients with a normal left ventricular function (ejection fraction >0.55; control population). All patients had an old (>4 weeks) myocardial infarction. By using age-adjusted peak VO2 values, a 'decreased' exercise capacity was defined as < or = predicted peak VO2 1 x SD (0.81 of predicted peak VO2), and a severely decreased exercise capacity as < or = predicted peak VO2 - 2 x SD (0.62 of predicted peak VO2). RESULTS: Patients in the study population (age 52+/-9 years; ejection fraction 0.46+/ 0.06) were mostly asymptomatic (NYHA class I: n=40, 76%), while 16 patients (24%) had mild symptoms, i.e. NYHA class II. All 17 controls (age 57+/-8 years) were asymptomatic. Mean peak VO2 was lower in patients with mild left ventricular dysfunction (23.6+/-5.7 vs 27.1+/-4.6 ml x min(-1) x kg(-1) in controls, P<0.05). In 75% of the study population patients (n=42) age-adjusted peak VO2 was decreased (NYHA I/II: n=29/13) and in 18% of them severely decreased (n=10; NYHA I/II: n=6/4). In contrast, only three patients (18%) in the control population had a decreased and none a severely decreased age-adjusted peak VO2. CONCLUSION: In patients with mild left ventricular dysfunction, who have either no or only mild symptoms of chronic heart failure, a substantial proportion has an impaired exercise capacity. By using age-adjustment, impairment of exercise capacity becomes more evident in younger patients. Patients with mild left ventricular dysfunction are probably under-diagnosed, and this finding has clinical and therapeutic implications. PMID- 9857923 TI - The optimal mode of coronary revascularization for diabetics. A risk-adjusted long-term study comparing coronary angioplasty and coronary bypass surgery. AB - AIMS: Some recent studies have reported-superior outcomes for diabetic patients following coronary bypass surgery compared with coronary angioplasty. However, the available data are conflicting, are based on relatively small numbers of diabetic patients, and have limited duration of follow-up. The aims of this study were to compare risk adjusted long-term survival in diabetic patients following first-time revascularization via either coronary bypass surgery or coronary angioplasty; and, to identify variables independently associated with mortality. METHODS AND RESULTS: This was a two centre database project involving 15809 patients undergoing either coronary angioplasty or coronary bypass surgery as their initial revascularization procedure. Diabetes was present in 1938 (12%). Mean follow-up was 4.6+/-2.7 years for angioplasty and 6.6+/-4.3 years surgery diabetic patients. Multivariable time-related analyses in the hazard function domain for death were performed. Overall ten-year survival for pharmacologically treated diabetics was better after coronary bypass surgery (60%) than angioplasty (46%, <0.0001). However, the risk-adjusted survival advantage conferred by bypass surgery over angioplasty was strongest for patients receiving oral agents for diabetic control (75% vs 62%) and less impressive for diet (84% vs 81%) and insulin-treated diabetics (63% vs 64%). The major factors independently associated with worse outcome after angioplasty were incomplete revascularization, and the use of a sulfonylurea agent. The use of the left internal mammary graft improved survival in surgical patients. CONCLUSIONS: In general, diabetic patients had better long-term survival after bypass surgery than angioplasty. Incomplete revascularization and sulfonylurea therapy worsened outcome after angioplasty, and use of the left internal mammary improved outcome after bypass surgery. PMID- 9857924 TI - A placebo-controlled study of growth hormone in patients with congestive heart failure. AB - AIM: Experimental data in heart failure models and an open trial of seven patients with idiopathic dilated cardiomyopathy have suggested beneficial effects of growth hormone on cardiac function. The aim of the present study was to evaluate growth hormone effects on cardiac function in a placebo-controlled study. METHODS: Twenty two patients with congestive heart failure of different aetiologies in NYHA II and III and an echocardiographic ejection fraction <0.45 were studied in a 3 month double-blind placebo-controlled study with growth hormone added to optimal heart failure therapy. Patients received either placebo (n=11) or recombinant human growth hormone (n=11) in an initial dose of 0.1 IU x kg(-1) week(-1) for 1 week, and thereafter 0.25 IU x kg(-1) week(-1) for the rest of the treatment period. Cardiac function was assessed by equilibrium radionuclide angiography and Doppler echocardiography. Functional capacity was evaluated by computerized bicycle exercise electrocardiography. RESULTS: Recombinant human growth hormone had no significant effect on systolic or diastolic cardiac function, exercise capacity or neuroendocrine activation. In addition, there was no overall improvement in functional class or dyspnoea grade. Insulin-like growth factor-I significantly increased demonstrating that the growth hormone had an endocrine effect. CONCLUSION: This is the first double blind and placebo-controlled study of the administration, over 3 months, of recombinant human growth hormone in patients with congestive heart failure of different aetiologies. The treatment was safe and without serious side effects. However, no beneficial effects on cardiac function or structure could be detected. PMID- 9857926 TI - Heart rate variability index in congestive heart failure: relation to clinical variables and prognosis. AB - AIM: To evaluate the clinical and prognostic value of the heart rate variability index in patients with congestive heart failure. METHODS: Sixty-four patients with chronic congestive heart failure and sinus rhythm underwent clinical assessment, 24-h ambulatory electrocardiography and echocardiography. Patients were followed for 6 to 30 months. Cardiac death or heart transplantation constituted the primary end-point of the study. RESULTS: The heart rate variability index was related to left ventricular ejection fraction (r=0.29, P=0.02) and New York Heart Association class (P=0.01). Patients with a restrictive left ventricular filling pattern had a lower heart rate variability index compared to patients with a non-restrictive pattern (26+/-11 vs 33+/-9 units, P=0.01). Patients who died (n=11) or underwent heart transplantation (n=4) had a lower heart rate variability index compared to survivors (21+/-10 vs 33+/-9 units, P<0.0001). In multivariate survival analysis, a reduced heart rate variability index was related to survival independent of parameters of left ventricular function. CONCLUSION: The heart rate variability index provides independent information on clinical status and prognosis in patients with chronic congestive heart failure. PMID- 9857925 TI - Safety and utility of atropine addition during dobutamine stress echocardiography for the assessment of viable myocardium in patients with severe left ventricular dysfunction. AB - OBJECTIVE: To assess the feasibility safety and side effects of the addition of atropine to dobutamine stress echocardiography for the detection of viable myocardium in patients with left ventricular dysfunction (ejection fraction < or = 35%) prior to coronary revascularization. BACKGROUND: The assessment of viable and/or ischaemic myocardium has high prognostic value as regards improvement of function and survival after coronary revascularization. The addition of atropine to dobutamine during echocardiographic testing for the presence of viable myocardium is not common practice. Consequently, no data exist on the safety and additional diagnostic value of this practice. METHODS: Two hundred patients with left ventricular ejection fraction < or = 35% were studied. RESULTS: Test end points were: target heart rate in 164 (82%) of the patients, severe angina in 18 (9%), maximum dobutamine-atropine dose in six (3%), severe ST segment changes in five (2%), cardiac arrhythmias in four (2%), and hypotension in three (1%). Viability could be assessed echocardiogaphically in 105/200 (53%) from a biphasic response (improvement of wall motion with low dose dobutamine and worsening with high dose), in 93 from ischaemia and in 12 from sustained or late improvements. In 36/105 (34%) patients, ischaemic myocardium could only be assessed after the addition of atropine. Cardiac arrhythmias occurred in 11/200 (6%) and hypotension (decrease of systolic blood pressure >30 mmHg) in 21/200 (11%). Neither the use of atropine nor the induction of ischaemia were associated with an increased incidence of cardiac arrhythmias or hypotension. CONCLUSIONS: In a large group of patients with severe left ventricular dysfunction, dobutamine stress echocardiography is feasible and safe in 186/200 (93%); the addition of atropine was necessary in 34% to assess myocardial viability. Hypotension and cardiac arrhythmias were the most frequent side effects, but were not related to the induction of ischaemia or addition of atropine. PMID- 9857927 TI - Mitochondrial DNA in idiopathic cardiomyopathy. AB - AIMS: To investigate the frequency of pathogenic mitochondrial DNA mutations in idiopathic cardiomyopathy. METHODS AND RESULTS: We investigated the occurrence of seven previously reported pathogenic mitochondrial DNA point mutations in 52 patients with idiopathic dilated cardiomyopathy (blood n=33, myocardium n=19), 10 patients with hypertrophic cardiomyopathy (blood n=7, myocardium n=3), 67 controls with ischaemic heart disease (blood n=53, myocardium n=14) and eight controls with no overt cardiac disease (blood n=4, myocardium n=4). Total DNA or cell lysates were studied by polymerase chain reaction amplification and restriction fragment length polymorphism analysis for the identification of the following mitochondrial DNA point mutations: A3243G, A3252G, A3260G, A4269G, A8344G, T8993G/C and T9997C. None of these point mutations were detected in the blood or myocardium of any of the individuals with dilated or hypertrophic cardiomyopathy or in the controls. In addition we investigated the occurrence of major deletions of mitochondrial DNA in eight patients with dilated cardiomyopathy (myocardium n=7, skeletal muscle n=1), three patients with ischaemic heart disease (myocardium n=3) and one control myocardium by Southern blot analysis. Deletions were not detected in any of the patients. CONCLUSION: The results suggest that although these mutations are known to be associated with specific cardiomyopathies, they are not a common feature of idiopathic cardiomyopathy. PMID- 9857928 TI - Papillary muscle shortening for mitral valve reconstruction in patients with ischaemic mitral insufficiency. AB - AIMS: To evaluate the feasibility of papillary muscle shortening in a specific group of high risk patients with ischaemic mitral regurgitation undergoing mitral valve reconstruction. BACKGROUND: From January 1996 to December 1997, 712 (10.1%) out of a total of 7042 open heart patients underwent mitral valve surgery in our hospital. Mitral valve reconstruction was performed in 408 of these patients (57.3%) and valve replacement had to be performed in 304 patients (42.7%). METHODS: A specific technique of papillary muscle reconstruction was performed in 32 patients undergoing valve reconstruction (7.8%). These cases had degenerated and had developed fibrotic elongated papillary muscles, which resulted in prolapses of one or more parts of the mitral valve leaflets. The aetiology in this group of patients was ischaemic, requiring concomitant myocardial revascularization in 28 patients (87.5%) with a mean of 2.7 grafts/patient. All patients underwent papillary muscle shortening using a pericardium pledget reinforced Polytetrafluoroethylene suture and annuloplasty with a Carpentier Edwards Physio Annuloplasty Ring. Of these 32 patients, 17 (53.1%) were male, the mean age was 67.1+/-9.7 years (range 41 to 81 years) and all but one were in pre operative NYHA class III or IV. RESULTS: There were two hospital deaths (6.2%). Postoperative Doppler echocardiography indicated satisfactory mitral valve function in all patients. Within the short mean follow-up period of 9.6+/-5.4 months (3 to 26 months) there was one non-cardiac-related death (3.1%). There was no need for reoperation, and no cases of thromboembolic and bleeding complications in the postoperative period. All patients were in NYHA functional class I or II at the time of follow-up. CONCLUSION: Our results indicate that mitral valve repair is a safe treatment for this group of high risk patients, and that papillary muscle shortening is a valuable tool in these patients with ischaemic mitral regurgitation undergoing surgery. PMID- 9857929 TI - Increased left ventricular systolic function in insulin dependent diabetic patients with normal albumin excretion. AB - Alterations in cardiovascular function may be an aetiological factor for the development of microalbuminuria in patients with insulin-dependent diabetes mellitus. We studied cardiac function with echocardiography in relation to the degree of albuminuria in 27 insulin-dependent diabetes mellitus patients and 13 healthy subjects. Patients were grouped according to urinary albumin excretion: <20 microg x min(-1) (normoalbuminuric), and 20 to 200 microg x min(-1) (microalbuminuric). None were or had been treated with cardiovascular drugs. The normoalbuminuric patients had a higher heart rate, mean velocity of circumferential shortening, stroke velocity index (a measure of contractility), and aortic peak velocity than controls. No difference in diastolic function was present. In the microalbuminuric group, the stroke velocity index was comparable to values observed in healthy subjects. The increased systolic performance (heart rate and contractility) may contribute to the renal hyperperfusion and glomerular hyperfiltration observed in insulin-dependent diabetes mellitus patients before the development of micro- and in turn macroalbuminuria. The possible cause effect mechanisms should be further studied, as preventive medical treatment of the hypercontractile heart is possible. In conclusion, cardiac contractility is increased in insulin-dependent diabetes mellitus patients with normoalbuminuria and returns to levels observed in healthy subjects when microalbuminuria develops. PMID- 9857930 TI - The NETWORK investigators. PMID- 9857931 TI - Biochemical markers in the diagnosis of coronary artery disease. AB - For many years cardiac markers have been used to classify whether chest pain is attributable to acute myocardial infarction or not. However massive, myocardial infarction is frequently preceded by plaque inflammation and local thrombus formation. Novel cardiac markers focus on detection of these more subtle manifestations of coronary artery disease. Detection of inflammation of coronary artery plaques is best achieved by measurement of C-reactive protein and fibrinogen, while thrombus formation may be assessed by testing for fibrin formation and platelet activation. When coronary flow is severely impaired minor myocardial injury will occur and cellular constituents may egress from damaged myocytes. Among the many cardiac markers for myocardial cell necrosis, troponin T revealed the highest sensitivity and cardio-specificity. The superior performance of troponin T has not only refined detection of myocardial cell necrosis but has also improved the risk stratification process and may even facilitate therapeutic decision making in patients with acute coronary syndrome. This review will summarize the characteristics and performance of diagnostic tools used for classification and risk stratification of patients with suspected myocardial injury. PMID- 9857932 TI - Standardization activities of markers of cardiac damage: the need of a comprehensive approach. AB - The field of biochemical markers of cardiac damage is in a dynamic state, with new applications continually appearing and new assays and markers being developed, but also with several analytical and interpretative problems to resolve. In this situation it is essential that a uniform and rigorous outlook be maintained to ensure optimal test utilization. For these reasons, the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) agreed to establish a Committee on 'Standardization of Markers of Cardiac Damage' (C-SMCD). The first task of the C-SMCD concerns coordination of the different activities from affiliated associations, with (1) a review of documents and recommendations and (2) supervision and, possibly, the take-over of the standardization activities. Second, the C-SMCD supports development of scientific programmes for forthcoming international meetings and congresses. The third task of the C-SMCD is development of specific additional projects to promote the rational use of cardiac markers and to identify new areas of research. PMID- 9857933 TI - Diagnostic strategies in myocardial infarction using myoglobin measurement. AB - Determination of myoglobin, a low molecular weight haeme protein (17.8 kDa), present in both cardiac and skeletal muscle, is an old test with new perspectives. Advantages and disadvantages of myoglobin determination are well known; recent availability of rapid and accurate methods for the assay of this protein has greatly enhanced interest in the clinical utilization of the test. However, since myoglobin is present in both skeletal and cardiac muscle, any damage to these muscle types results in its release in blood and, consequently, high levels of the protein are observed in conditions unrelated to acute myocardial infarction (AMI). New strategies for myoglobin measurement may resolve this limitation. These strategies include both the combined measurement of myoglobin and a skeletal specific marker (carbonic anhydrase III) or a cardiac specific marker (troponin I or T), as well as myoglobin evaluation on serial samples. In particular. the diagnostic algorithm based on combined measurement of myoglobin and troponins significantly improves diagnostic efficiency in laboratory assessment of suspected AMI patients. However, further efforts are necessary to improve the standardization of present methods for myoglobin measurement: the significant disagreement demonstrated between some commercially available assays should be carefully considered in clinical practice. PMID- 9857934 TI - Troponin T or troponin I or CK-MB (or none?). AB - Differential diagnosis of patients who present with chest pain remains problematical. It has been shown that 11.8-7% of patients with acute myocardial infarction (AMI) are sent home from the emergency department (ED). Audit of our own ED has shown the incidence of missed prognostically significant myocardial damage to be 6.7%. Diagnostic criteria for AMI have classically been based on the triad of history, ECG and measurement of cardiac enzymes. The choice of 'cardiac enzymes' has been dictated by the evolution of laboratory techniques, commencing with measurement of aspartate transaminase and progressing to measurement of creatine kinase (CK) and its MB isoenzyme (CK-MB). Measurement of CK-MB has been shown by both clinical studies and rigorous statistical analysis to represent the best test for the diagnosis of AMI. The advent of real time immunoassay together with advances in therapeutic options for management of acute coronary syndromes (ACS) has resulted in a paradigm shift in the approach to laboratory testing. Immunoassay for CK-MB (CK-MB mass measurement) is diagnostically superior to CK MB activity measurement and is the test of choice for 'classical' AMI. Development of immunoassays for the cardiac troponins, i.e. cardiac troponin T (cTnT) and cardiac troponin I (cTnI), has enhanced diagnostic specificity. These measurements are completely specific for cardiac damage, allow quantitation of the extent of infarction and are diagnostically superior to CK-MB measurement. Applications of this specificity have included the differential diagnosis of CK elevation in arduous physical training, detection of myocardial damage after DC cardioversion and prediction of ejection fraction. Of more interest is the utility of these markers in management of patients presenting without clear electrocardiographic changes. Diagnosis and management of patients presenting with ST segment elevation has been clarified by large clinical trials of thrombolytic agents. In such patients, thrombolysis is the treatment of choice. Patients presenting with ST segment elevation represents the minority of patients with probable ACS 9.6% of all patients presenting to our hospital. The majority require risk stratification into high- and low-risk groups. It is here that cardiac troponins have a major role. The measurement of cTnT has been shown in a large number of studies to enable risk stratification of patients with unstable angina. The combination of cTnT, admission ECG and stress ECG can be used for a comprehensive risk stratification of patients with unstable angina. The combination of cTnT, admission ECG and stress ECG can be used for a comprehensive risk stratification which can be completed by 24 h from admission, as well as allowing a safe discharge policy from the ED. Measurements of cardiac troponins can also be used to predict prognosis in patients with other diagnostic categories. Patients with cardiac failure can be risk stratified according to cTnT status. cTnT status on admission allows subdivision into high- and low-risk groups in patients presenting with ST segment elevation. Certainly, cTnT measurement can be incorporated into a clinical decision-making strategy to assign patients to investigation and management pathways. There is evidence that cTnT may be useful to guide therapeutic options. The major issue is one of cost. In the U.K. model of managed care with undemanding diagnostic standards, the role of cTnT will be to enhance clinical decision-making strategies, to provide accurate diagnosis and to reduce lengths of stay. This can be shown to have potential for major improvements in cost efficiency. Improvements in diagnostic accuracy can reduce inappropriate long-term drug therapy. In systems with a more aggressive laboratory investigation strategy, rationalization of test numbers will provide an immediate cost reduction while improving quality. Finally, use of point-of-care testing (POCT) means that biochemical testing can be pe PMID- 9857935 TI - Biochemical differences between cTnT and cTnI and their significance for diagnosis of acute coronary syndromes. AB - Although assays for cardiac troponin T (cTnT) and cardiac troponin I (cTnI) exhibit similar clinical performance in patients with acute coronary syndromes for diagnosis and risk stratification, there are differences in the release and clearance of these proteins from damaged myocytes. There are also differences in the percentages of patients with chronic renal failure and skeletal muscle myopathies that have abnormal results. A review of the biochemistry of these proteins may provide insights as to the nature of these differences. Cardiac troponin T has a higher overall tissue concentration and free cytoplasmic concentrations than cTnI, and appears in blood of patients with acute myocardial infarction as a mixture of complexed (cTnT-I-C) and free cTnT. Cardiac troponin I is more hydrophobic and appears in blood predominately in the binary complex (I C) form, with smaller amounts of the ternary (T-I-C) complex. Degradation of the complex will lead to faster clearance of the protein. There are also differences in standardization and availability of commercial troponin kits. Cardiac troponin T assays are available from only one manufacturer and are standardized to a single material, whereas cTnI is available from multiple vendors. While there is some degree of correlation between assays, there are significant proportional biases between results which can be problematical when different tests are made on the same patient. PMID- 9857936 TI - Expression of cardiac troponin T isoforms in skeletal muscle of renal disease patients will not cause false-positive serum results by the second generation cardiac troponin T assay. AB - The purpose of this study was to determine whether the two monoclonal anti cardiac troponin T (cTnT) antibodies used in the second generation cTnT assay (capture Ab, M11.7; detection Ab, M7) would detect expression of cTnT isoforms in skeletal muscle from chronic renal disease patients. Skeletal muscle biopsies obtained from 45 chronic renal disease patients (as well as human heart muscles and normal human skeletal muscles) were prepared for Western blot analysis and blotted with the following anti-cTnT antibodies: M 11.7; M7; JS-2, Lakeland Biomedical; 13-11, Duke University) and anti-cTnI antibody JS-1. Using the M11.7 Ab, 20 of 45 renal skeletal muscles demonstrated one to three cTnT isoforms, MW34 39 kDa. These findings were confirmed by both the Lakeland and Duke antibodies. However the BM M7 antibody detected, in two of 45 muscles, only a protein with a MW of approximately 39 kDa. All four antibodies demonstrated equivalence in detection the 39 kDa cTnT isoform expressed in heart muscle. None detected any isoforms in normal skeletal muscle. A single cTnI isoform, MW 25 kDa, was detected by JS-1 only in normal adult myocardium. Based on the antibody configuration of the second generation cTnT assay, we conclude that while cTnT isoforms are expressed in human skeletal muscle obtained from chronic renal disease patients, if released into the circulation, they would not be detected. PMID- 9857937 TI - Troponin T, a predictor of death in chronic haemodialysis patients. AB - Of the 10 chronic haemodialysis patients whose serum TnT levels exceeded the threshold value of 0.1 microg.L(-1) at entry into the study, four were dead at 1 year and three others had a diagnosis of CAD. Of the 20 chronic haemodialysis patients with normal serum TnT levels at entry, one died and none had CAD. All five deaths were cardiac related, either arising from acute myocardial infarction or by sudden death. When serum TnT levels were compared with accepted predictors of death in chronic haemodialysis patients, such as serum creatinine, serum albumin and haematocrit, in the present study serum TnT proved to be more accurate and had excellent sensitivity and specificity. Serum TnT was also superior to serum TnI, which proved to be no more discriminating than the non specific muscle marker, aldolase. PMID- 9857938 TI - Experience with cardiac troponin T in difficult cases. AB - Recent years have seen the advent of new cardiac markers. These have been shown to be valuable in large controlled clinical trials. The breakthrough of the troponins world-wide for detection of myocardial lesions has meant that the Cardiac Troponin T parameter has been available in our hospital since August 1996. Analysis of the samples is carried out on the Elecsys, an automated immunology analyzer from Roche Diagnostics, Austria. The purpose of the present paper is to investigate whether study results on the diagnostic utility, sensitivity and specificity of troponin T can be confirmed in multimorbid patients in routine clinical practice, who are otherwise excluded from such trials. Troponin T was investigated in five cases including patients presenting with myocarditis and parallel damage of the skeletal muscles, as well as in haemodialysis patients. Cardiac troponin T has proved to be a very reliable parameter in all of these patients due to its high sensitivity and specificity, where other conventional diagnostic procedures were not helpful due to co morbidity and confounding factors. New clinically relevant information can be obtained by both individual as well as serial measurements. Cardiac troponin T determination is a powerful new tool and should be considered seriously, regardless of other underlying diseases: it is, therefore, not only a diagnostic but also a management tool. PMID- 9857939 TI - Diagnostic accuracy, angiographic correlates and long-term risk stratification with the troponin T ultra sensitive Rapid Assay in chest pain patients at low risk for acute myocardial infarction. AB - AIMS: This prospective study of acute chest pain patients clinically at low risk for a myocardial infarction was designed to: determine the diagnostic accuracy of a cardiac troponin T (cTnT) ultra sensitive Rapid Assay (RAII) compared with the quantitative cTnT assay; evaluate the association of a positive RAII with the presence and severity of coronary artery disease (CAD); and determine the ability of the RAII result to predict adverse events during long-term follow-up. METHODS AND RESULTS: A total of 199 patients referred for chest pain, without ST segment elevation on presenting ECG, underwent RAII, quantitative cTnT, CK and CK-MB tests drawn simultaneously > or = 10 h after symptom onset. An abnormal value for cTnT was defined as >0.1 ng.mL(-1). The presence and extent of CAD was recorded in patients undergoing angiography. Adverse events, including cardiac death, non fatal infarction, and readmission for unstable angina or heart failure, were assessed long-term. An abnormal RAII was found in 41 (20-6%) patients. The RAII sensitivity for detecting abnormal quantitative cTnT levels was 100%, specificity 96.3% (158/164) and overall concordance 97.5%. Although the presenting ECG was normal or non-specific in 95%, ST depression or T wave inversion occurred in 17% of RAII-positive versus 2%, RAII-negative patients (P=0.004). Of RAII-positive patients who underwent angiography (79%), 87% had CAD and 60% had multivessel disease. Kaplan Meier event-free survival curves showed early separation and continued to modestly diverge for patients with positive and negative RAII (69% versus 90% one-year event-free survival, P=0.002). CONCLUSION: In a chest pain population anticipated to have a low prevalence of acute coronary syndromes and a good prognosis, the RAII is a quick and reliable test. It provides an important initial opportunity to identify patients with a high prevalence of CAD and increased incidence of future cardiac events. PMID- 9857940 TI - Progress in the diagnosis of unstable angina and perspectives for treatment. AB - Unstable angina is a high-risk phase of coronary heart disease defined by clinical symptoms. Electrocardiographic findings are heterogeneous and creatine kinase is only rarely elevated. However, troponin T and troponin I are found in about 30-40% of patients with angina at rest disclosing minor myocardial cell injury. Several studies in recent years have documented that troponins are the best markers to identify high-risk patients when other life-threatening, non cardiac diseases are excluded. In addition, first evidence is provided that only these patients benefit from prolonged treatment with low molecular weight heparin or glycoprotein IIb/IIIa receptor blockers. Therefore, this new diagnostic potential should be made available to emergency rooms for risk stratification and in the future to guide therapeutic decision making. PMID- 9857941 TI - The FRISC experience with troponin T. Use as decision tool and comparison with other prognostic markers. AB - Early prognostic evaluation is essential for the application of appropriate treatment and further management in the heterogeneous population with unstable coronary artery disease. Among the variety of prognostic indicators that have been suggested, biochemical markers of myocardial damage, especially the troponins, have gained increasing interest. The FRISC troponin T (TnT) substudy has validated and provided additional insights into the use of biochemical markers for prognostic evaluation. Nine hundred and seventy-eight patients were enrolled and followed for 5 months. An increase in the rate of cardiac death or myocardial infarction was seen during follow-up, with increasing levels of TnT. The TnT level separated low and high risk patients better than the conventional division of unstable angina and myocardial infarction. In the comparison with other early available risk indicators TnT was an independent risk indicator and gave additive prognostic information to both inclusion ECG and the result of a predischarge exercise test. TnT was superior to CK-MB for risk stratification, while TnT and troponin I showed comparable results. In patients with elevated TnT, treatment with dalteparin significantly decreased the risk for death or myocardial infarction compared to placebo, while there was no benefit from dalteparin in patients without Tn-T elevation. Based on the experiences of the FRISC study and the literature we suggest that the use of troponins should be included in the diagnostic and prognostic evaluation of patients with suspicion of unstable coronary artery disease. PMID- 9857943 TI - Cardiac rehabilitation: where are we going? AB - Several components of cardiac rehabilitation can be considered to be of proven value: smoking cessation reduces cardiac events by 50% in observational studies, dietary modification such as the so-called Mediterranean diet--if rigorously applied--reduces cardiac events, decreases progression and has favourable effects on hypertension, osteoporosis and the risk of cancer. Cholesterol lowering by HMG CoA reductase inhibitors is cost effective but not practiced widely enough. Exercise training programmes improve functional capacity and symptoms particularly in patients with compensated heart failure. The prognostic implications are unclear at present. Stress modification is useful and return to work may be enhanced by cardiac rehabilitation. Cardiac rehabilitation, as a multifactorial intervention prevents progression, improves well being and prognosis, and should be an integral part of the cardiological management after a cardiac event. PMID- 9857942 TI - The cardiac troponins: uses in routine clinical practice. Experiences from GUSTO and other clinical trials. AB - Recent advances in pharmacological and mechanical approaches to acute coronary syndromes have led to rapid changes in the management of patients admitted with acute coronary syndromes. These changes have been mirrored by the appearance of newer highly specific biochemical markers of myocardial damage particularly the cardiac troponins. When new biochemical markers become available it is the responsibility of the clinical chemist to evaluate them critically in terms of sensitivity, specificity, efficiency and analyzer precision, in the rigid setting of quality control that laboratories practise, and to compare them with other markers. When the data are shown to Clinical Cardiologists with supporting statements such as 'useful management tool' and 'can be used for early diagnosis of Myocardial Infarction', a different set of questions may need to be answered. The 'So what?' response is most frequent and is the most important hurdle that these newer biochemical markers have to overcome to convince physicians to change their current practice. This presentation will review the results of studies that have examined the potential clinical usefulness of the cardiac troponins with respect to diagnosis and risk stratification of patients admitted with suspected acute coronary syndromes. Any troponin variable that survives the 'so what' question has one further major hurdle to overcome. This is the requirement to inform physicians what different therapeutic strategies they should follow if the variable is present. Available clinical trial evidence about differing management options for patients according to their troponin status will be reviewed and outline management algorithms will be presented. Many questions remain unanswered and these will be included at the time points where they may be relevant. PMID- 9857944 TI - Diagnosis and management of new cardiovascular risk factors. AB - Independent risk factors for coronary heart disease (CHD) can best be identified by means of long-term prospective epidemiological studies. These factors should not be viewed in isolation, but as part of a complex governing a person's global risk of CHD. In the Munster Heart Study (PROCAM), 8-year follow up of a cohort of middle-aged men has led to the identification of nine variables which independently contribute to CHD risk: age, smoking history, personal history of angina pectoris, family history of myocardial infarction (MI), presence of diabetes mellitus, systolic blood pressure and the levels of low density lipoprotein (LDL) cholesterol, high density lipoprotein cholesterol and triglyceride. An algorithm based on these risk factors may be used to calculate an individual's risk of fatal or non-fatal MI and is available in interactive fashion on the website of the International Task Force for Prevention of Coronary Heart Disease at www.chd-taskforce.com. Treatment goals for LDL cholesterol depend on a person's level of risk: for persons with a small increase in risk 160 mg x dl(-1), for those with a moderate increase in risk 135 mg x dl(-1) and for those at high risk, including those with a history of CHD (secondary prevention) 100 mg x dl(-1). Intervention trials indicate that lowering of LDL cholesterol for 5 years produces much of the benefit predicted from the epidemiological data. The place of newer risk markers such as Lp(a), homocysteine, and parameters of clotting and inflammation in risk prediction and management remains to be determined. PMID- 9857945 TI - Mental stress and ischaemic heart disease: an underestimated connection. AB - In this paper a theoretical framework is offered for mental stress as a risk factor for the development of ischaemic heart disease (IHD). The development of IHD related to mental stress can be described as dependent on interaction between the following factors: (1) the presence of stressors, (2) activation of a receptor and transformation system (i.e. the central nervous system) and (3) physiological effector systems. Stressors are defined as external or internal, nomothetic or idiosyncratic and are exemplified. A prerequisite for the psychological stress reaction is the existence of a central nervous system in which stimuli are perceived, interpreted and responded to. The interaction between neocortex and paleocortex is discussed as well as the initiation of the cerebral stress reaction. Four physiological effector systems seem to be related to mental stress: (1) the autonomic nervous system (ANS), (2) the hypothalamic pituitary-adrenal (HPA) axis, (3) the peripheral nervous system (PNS) and (4) the endorphin system. In the development of cardiovascular diseases the pathophysiological changes in the ANS and in the HPA axis play the most important roles. PMID- 9857946 TI - Interventional cardiology in perspective: impact on cardiac rehabilitation. AB - Exponential evolution of percutaneous, catheter-based revascularization has modified traditional premises of cardiac rehabilitation. With increasing numbers of percutaneous transluminal angioplasty (PTCA) procedures relative to coronary artery bypass grafting interventions, the need for in-patient rehabilitation is declining; on the other hand, the need for life-style counselling within the framework of out-patient rehabilitation is increasing. Risk factor interventions are important to minimize and to retard the progression of the disease. Rehabilitation programmes offer exercise training, stress modification and life style counselling to reduce cardiovascular risk factors and to increase exercise capability. Since costs of out-patient rehabilitation seem to be lower than those of in-patient rehabilitation, a case may be made in favour of directing limited funds to additional interventions and to saving money by replacing in-patient rehabilitation with outpatient programmes after PTCA. The budget-holder has to make the choice. PMID- 9857947 TI - Optimizing exercise training for subgroups of patients with chronic heart failure. AB - Patients with heart failure and severe left ventricular dysfunction are limited in their ability to tolerate exercise. Recent research has demonstrated that it is not a restricted cardiac output which limits exercise but an abnormality in the performance of a more peripheral component in the pathway of delivery of oxygen to the exercising muscles. Deficiencies in peripheral blood flow and skeletal muscle function and an exaggerated sensitivity to exercise-derived metabolic signals lead to early and profound exercise-induced fatigue and dyspnoea, the mechanisms of which we are only now beginning to understand. These findings raise the possibility of improving exercise tolerance by peripherally acting therapies such as exercise training of skeletal muscles in left ventricular dysfunction and chronic heart failure. Training has been shown to improve exercise tolerance at least as much as angiotensin-converting enzyme inhibitors in these syndromes; additionally, training improves ventilatory control, skeletal muscle metabolism and autonomic nervous system activity. As yet, the most appropriate forms of exercise therapy are not known nor is how best to choose the most suitable patients for this form of treatment. There is evidence that patients with at least moderate, and possibly severe but stable, heart failure can benefit from exercise rehabilitation, provided it is tailored to the capacity of the patient and certain safety considerations are taken into account. PMID- 9857948 TI - Optimal timing of phase II rehabilitation after cardiac surgery. The cardiologist's view. AB - Structured programmes of rehabilitation have been shown to be effective in restoring functional capacity, increasing return to work and improving the psychosocial status of patients following open heart surgery. However, uncertainty still exists about the optimal timing of rehabilitation after cardiac surgery. Data in the literature suggest that phase II rehabilitation can begin as early as one week after open heart surgery without having a negative influence on infections, mortality or readmissions. Early rehabilitation can even improve graft patency after CABG. However, age, operative complications and co morbidities must be considered individually for each patient when determining the beginning of phase II rehabilitation. PMID- 9857949 TI - Optimal timing of rehabilitation after cardiac surgery: the surgeon's view. AB - Cardiac rehabilitation has become an accepted adjunct treatment for the majority of patients with cardiovascular disease, especially for those who have received cardiac surgery. However, improved survival has not been generally demonstrated in a supervised cardiac rehabilitation programme, while some benefits have been found in functional capacity, psychosocial characteristics and lipoprotein patterns of patients who underwent sustained periods of exercise training. The influence of post-surgical conditions on phase II rehabilitation following cardiac surgery has not yet been well addressed. Many factors may influence the timing of phase II rehabilitation following cardiac surgery, especially the pre operative condition of the patient, the concomitant morbidity, the incidence of peri-operative complications and the rapidity of recovery, mainly influenced by post-operative cardiac and lung function, pain and wound healing. This article reviews the general and specific medical and surgical problems encountered during early follow-up of cardiac surgery patients, which might affect the timing of postoperative rehabilitation, analyses briefly the impacts of less invasive heart surgery on cardiac rehabilitation as well as that of fast tract protocols after conventional heart surgery. Some patients required service intervention during cardiac rehabilitation while others were withdrawn from cardiac rehabilitation for medical reasons or surgery-related complications. Specific problems in patients following cardiac transplantation are depicted briefly. Cardiac transplant recipients may suffer from pre-operative end-stage heart failure, prolonged cold ischaemia of the donor heart, denervation of the cardiac allograft, immunological allomismatch, and unusual psychological stress. In summary, phase II cardiac rehabilitation on a stationary and more recently on an ambulatory basis is generally recommended two to four weeks following uncomplicated coronary and valvular procedures, while patients following cardiac transplantation may be included in such programmes after approximately four to six weeks. The earliest rehabilitation is possible in patients following less invasive heart surgery and may start one to two weeks postoperatively. PMID- 9857950 TI - Comprehensive cardiac rehabilitation: is it cost-effective? AB - A major challenge for all health care systems is to identify the most efficient use of limited and finite resources available for health care. Economic evaluation provides a balance sheet of the benefits, harms and costs for making choices between alternative health care services and is one strategy to assist decision-makers to make rational choices about effective and efficient health care. Cost and outcomes data collected on two or more alternatives form the basis for economic evaluations and calculating a cost-effectiveness ratio. While comprehensive cardiac rehabilitation has been shown to be an effective intervention for patients with documented heart disease, the economic evaluation data from which to determine the efficiency of cardiac rehabilitation are limited. Available economic evaluations of comprehensive cardiac rehabilitation suggest that it is a cost-effective intervention following an acute coronary event that can be economically justified. Although the majority of data from studies with less rigorous designs suggest either savings or a decrease in health care utilization, there were increased costs per quality-adjusted life year gained in the only randomized controlled trial with a cost-effectiveness analysis of cardiac rehabilitation. As the traditional delivery of cardiac rehabilitation services is undergoing re-examination, there is a need for considerably more research on the cost-effectiveness of cardiac rehabilitation before any definitive statement about reimbursement is made. PMID- 9857951 TI - Recovery of high potassium-evoked dopamine release after depolarization challenge in the striatum of young and old male rats. AB - The aim of this study was to assess the recovery of high potassium-evoked dopamine (DA) release after depolarization challenge in young (3-4 months) and old (21-25 months) male Wistar rats. Recovery of DA release was evaluated by comparison of the peak responses of DA release induced by two serial high potassium stimulations. Concentric microdialysis probes were stereotaxically implanted in the lateral striatum of rats, and microdialysis was commenced 24 h after surgery. Using a low flow rate of perfusion (1 microl/min), all rats received 2 x 20 min infusions of 100 mM potassium solution separated by either 60 or 140 min. No difference in the basal DA concentration or the potassium-evoked DA release or its recovery was seen between the two groups. Our results suggest that the vesicular DA store recovers rapidly after high potassium challenge in both young and old rats. PMID- 9857952 TI - Activity of tachykinin NK1 and bradykinin B2 receptor antagonists, and an opioid ligand at different stimulation parameters in neurogenic inflammation in the rat. AB - Stimulation of the saphenous nerve in the anaesthetised rat results in cutaneous neurogenic oedema formation. We have examined the effect of a tachykinin NK1 and a bradykinin B2 antagonist, and a mu-opioid agonist on plasma extravasation observed in response to two differing nerve stimulating parameters (10 V, 1 ms, 2 Hz and 25 V, 2 ms, 10 Hz). The NK1 antagonist SR140333 abolished oedema, supporting the theory that an NK1 agonist is a primary mediator of neurogenic oedema. The B2 antagonist HOE 140 had no effect, indicating a lack of involvement of B2 receptors in this response. The pre-junctionally acting mu-opioid agonist DAMGO significantly inhibited oedema formation at the 10 V, 1 ms, 2 Hz (P < 0.001), but not the 25 V, 2 ms, 10 Hz stimulation parameters. Thus a post junctionally acting NK1 antagonist inhibited neurogenic oedema formation induced by both stimulation parameters, whilst a pre-junctionally acting mu-opioid agonist acted only at 10 V, 1 ms, 2 Hz parameters. These findings could be of interest with respect to therapeutic approaches of pathophysiological conditions which involve a neurogenic component. PMID- 9857953 TI - The onset of diffuse noxious inhibitory controls in postnatal rat pups: a C-Fos study. AB - The development of diffuse noxious inhibitory controls (DNIC) was studied in postnatal rats aged 12, 21 and 42-days-old, using immunoreactive localization of the c-fos protein products induced in the dorsal horn by noxious stimulation. The presence of DNIC was revealed by the reduction in the levels of Fos-like immunoreactivity that are normally induced by a standardized primary pinch stimulus when this stimulus was accompanied by a concurrent noxious stimulus (formalin) to a heterotopic body part. Significant reductions were seen at postnatal day 42 (P42; 15% reduction) and P21 (17% reduction), but concurrent stimulation had no significant effect at P12. These results suggest that the system subserving DNIC is functionally mature by P21, but not effective at P12. This delayed maturation of an inhibitory system may underlie the extreme sensitivity to somatosensory stimulation seen in neonatal pups and premature infants. PMID- 9857955 TI - 1-Trichloromethyl-1,2,3,4-tetrahydro-beta-carboline increases extracellular serotonin and stimulates hydroxyl radical production in rats. AB - 1-Trichloromethyl-1,2,3,4-tetrahydro-beta-carboline (TaClo), a neurotoxin structurally similar to the dopaminergic neurotoxin MPTP, may be formed in humans treated with chloral hydrate or exposed to trichloroethylene, a widely used industrial solvent. Systemically administered TaClo (0.4 mg/kg, i.p.) induced an immediate and transient release of dopamine (DA) and serotonin (5-HT) measured using microdialysis. However, only 5-HT was increased significantly (area under the curve, AUC, for the 1-2 h-period following TaClo administration: 400% compared with the respective control value; 2-3 h-period: 326%). This was followed by a progressive increase in hydroxyl radical formation reflected by higher extracellular concentrations of the hydroxylate product of salicylic acid, 2,3-dihydroxybenzoic acid (AUC for the 1-2 h period following TaClo administration: 182% compared with the respective control value; 2-3 h period: 190%). In contrast, extracellular glutamate and GABA were increased 2-3 h post injection by 64 and 51%, respectively. These data suggest that TaClo stimulates the generation of hydroxyl free radicals via an acute release of 5-HT and perhaps DA. PMID- 9857954 TI - Anticonvulsant effects of melatonin in amygdala-kindled rats. AB - Different doses of melatonin (50-100 mg/kg i.p.) were tested for their ability to suppress complex partial seizures in the amygdala kindling model of the rat. Thirty minutes after injection of 75 or 100 mg/kg melatonin, the current threshold necessary to elicit epileptic afterdischarges (ADT) was significantly increased by about 200 or 250%, respectively. These doses were also sufficient to suppress generalised seizures at threshold current. All doses of melatonin decreased body temperature by more than 0.5 degrees C, but caused no pronounced ataxia. Seizure susceptibility in terms of ADT changed with the time of the day but seems not to depend on the circadian variation of endogenous melatonin, since the highest ADT was found in the morning when endogenous melatonin levels are low. PMID- 9857956 TI - Relationship between neuronal activity and substance P-immunoreactivity in the rat spinal cord during acute and persistent myositis. AB - The spinal level of substance P (SP) is assumed to be an important determinant of neuronal activity under pathophysiological conditions. In rat dorsal horn neurones, impulse activity was studied during a carrageenan-induced acute (2-8 h) and a Freund's adjuvant-induced persistent (12 days) myositis and compared with the spinal substance P-immunoreactivity (SP-IR) of the same animals. Myositis induced changes in responsiveness of the neurones reached a maximum within 2-8 h, whereas background activity of the neurones was highest after 12 days of myositis. The area of SP-IR in the superficial dorsal horn decreased during acute and persistent myositis and the integrated density of the staining was largely unchanged. The difference in time-course between neuronal activity and SP-IR suggest that during persistent myositis factors other than SP gain more influence on the behaviour of the neurones. PMID- 9857957 TI - Transcranial laser doppler mapping of activation flow coupling of the rat somatosensory cortex. AB - Signal averaged laser Doppler (LD) through a thinned skull over the rat somatosensory cortex was used to map the spatial and temporal characteristics of activation-flow coupling, the change in regional cerebral blood flow (CBF) due to neuronal activation, in response to electrical forepaw stimulation. The location of maximal changes in amplitude of the LD response was reproducibly recorded at 4 5 mm lateral to Bregma. This location is very similar but slightly posterior and lateral to results obtained from electrophysiological, magnetic resonance imaging (MRI), and optical imaging studies. The latency of the activation-flow coupling (AFC) response was inversely correlated to response amplitude, with shorter latencies at positions of maximal amplitude. PMID- 9857958 TI - Formation of granular cytoplasmic aggregates in COS7 cells expressing mutant Cu/Zn superoxide dismutase associated with familial amyotrophic lateral sclerosis. AB - To investigate the molecular mechanisms of the 'gain of toxic function' of mutant Cu/Zn superoxide dismutase (SOD) associated with familial amyotrophic lateral sclerosis (FALS), mutant (Ala 4 --> Thr, Gly 85 --> Arg, Gly 93 --> Ala, and two base-pair deletion in the 126th codon), as well as wild-type (wt), Cu/Zn SODs were expressed in COS7 cells. The formation of granular cytoplasmic aggregates accompanied by collapse of the cytoplasm was observed in cells expressing mutant (mt) Cu/Zn SODs, but not in cells expressing wt Cu/Zn SOD. The aggregates contained ribosome-like particles and endoplasmic reticulum. These results suggest the possibility that mt Cu/Zn SODs promote the formation of aggregates which are toxic to cells. PMID- 9857959 TI - Removal of superficial inhibition releases hyperexcitability in middle and deep horizontal slices from rat somatosensory neocortex. AB - Synaptic physiology was studied in horizontal slices of rat somatosensory neocortex. Intrinsic properties of pyramidal neurons from horizontal slices resembled those recorded in cells from coronal slices, but cells in superficial horizontal slices displayed more prominent fast and slow inhibition, while cells from deeper slices showed disinhibition. This disinhibition in deeper horizontal slices resulted in epileptogenesis in 81% of middle and 35% of deep layer horizontal slices. Brief exposure to glutamate antagonists and dantrolene was ineffective in preventing epileptic activity, but limited pentobarbital exposure reduced the proportion of deep slices manifesting epileptiform activity by 75%. Thus, within cortex inhibition dominates superficially and excitation predominates in deeper layers. While the cortex is vulnerable to hyperexcitability when superficial cortex is compromised, enhancing fast inhibition can reset the excitation-inhibition balance, and prevent epileptogenesis. PMID- 9857960 TI - Differential distribution of alternatively spliced somatostatin receptor 2 isoforms (sst2A and sst2B) in rat spinal cord. AB - We have recently shown that the cytoplasmic tail of the somatostatin receptor sst2 undergoes alternative splicing giving rise to two isoforms, SSt2A and sst2B. In the present study, we have raised polyclonal antibodies that specifically detect either sst2A or sst2B and used these antisera for immunocytochemical localization of the receptor proteins in the rat spinal cord. sst2A immunoreactivity formed a dense network consisting of neuronal perikarya and dendrites in the superficial layers of the dorsal horn. In contrast, prominent sst2B-immunoreactivity was found on neuronal perikarya and proximal dendrites throughout the gray matter of the spinal cord. Taken together, we show that alternative carboxy-terminal splicing is involved in cell-specific expression of somatostatin receptor sst2 isoforms in rat spinal cord, and that sst2A and sst2B mediate effects of somatostatin at different cellular sites. PMID- 9857961 TI - Theta synchronization in the human EEG and episodic retrieval. AB - Event-related desynchronization (ERD) and synchronization (ERS) was measured during episodic retrieval in a specially designed recognition task which forces subjects to avoid semantic search strategies. ERD represents the percentage of a decrease, ERS an increase in band power. The results show that only the theta band differentiates between good and bad episodic memory performers and that good performance is related to a large degree of theta synchronization. The delta and alpha bands did not yield significant effects. Topographical differences in theta ERS reveal that good performers use primarily their right hemisphere to retrieve episodic information. This finding agrees with respective results from PET studies. PMID- 9857962 TI - Association between polymorphisms in the type 1 sigma receptor gene and schizophrenia. AB - Several antipsychotic agents such as haloperidol and rimcazole are known to bind to sigma receptors with high affinity, and evidence for a potential link between sigma receptors and the etiology of schizophrenia has been reported. The present study was conducted to systematically search for nucleotide variants of the type 1 sigma receptor gene in 48 schizophrenics. Two polymorphisms were found: GC-241 240TT in the 5' flanking region and Gln2Pro. These two polymorphisms were in nearly complete linkage disequilibrium with each other. The Pro2 variant of the Gln2Pro polymorphism changes the endoplasmic reticulum retention signal motif. These polymorphisms were examined in an extended sample of schizophrenics (n = 308) and controls (n = 433) and a significant association between the presence of the TT/Pro2 haplotype and schizophrenia was observed (odds ratio = 1.27, P = 0.04). PMID- 9857963 TI - A novel Drosophila paired-like homeobox gene related to Caenorhabditis elegans unc-4 is expressed in subsets of postmitotic neurons and epidermal cells. AB - We have isolated a novel Drosophila paired-like homeobox gene, DPHD-1. The homeodomain of DPHD-1 showed 85% amino-acid identity with that of the C. elegans Unc-4 protein. Whole-mount in situ hybridization of embryos and third-instar larvae revealed that the DPHD-1 mRNA is specifically localized in subsets of postmitotic neurons in the central nervous system (CNS) and in the developing epidermis with a segmentally repeated pattern. Double staining with a posterior compartment marker, an anti-Engrailed antibody, showed that DPHD-1 expressing neurons in the CNS were present in the posterior compartment, whereas DPHD-1 expression in the epidermis was restricted to the anterior compartment in each segment. This temporal and spatial expression pattern suggests that DPHD-1 may play a role in determining the distinct cell types in each segment. PMID- 9857964 TI - Chronic elevation of amyloid precursor protein expression in the neocortex and hippocampus of rats with selective cholinergic lesions. AB - Overexpression or aberrant processing of the beta-amyloid precursor protein (APP) and loss of cortical cholinergic function represent two hallmark pathological features of Alzheimer's disease, although it is still unclear whether these alterations take place independently or in an inter-related manner. In the present study, the possible relationships between altered APP expression and cholinergic hypofunction in the neocortex and hippocampus were addressed histologically following selective and complete (90-95%) removal of the basal forebrain cholinergic neurons by the 192 IgG-saporin immunotoxin, at a dose (5.0 microg, intraventricularly) producing profound and permanent cognitive deficits. Computer-aided densitometric analyses revealed, at 6 months post-lesion, a virtually complete loss of terminal cholinergic innervation in various neocortical and hippocampal regions (up to 80%), which correlated highly with the marked (up to 71%) increases in APP expression measured in the same territories. The present results indicate that the integrity of ascending basal forebrain cholinergic inputs to the neocortex and hippocampus may be required for the maintenance of physiological levels of APP expression in the same regions, thus providing a novel rationale for interventions aimed at restoring or enhancing cortical cholinergic neurotransmission. PMID- 9857965 TI - Glibenclamide, an antagonist of ATP sensitive K+ channels, blocks free radical generation in the rat myocardium. AB - The present study examined the effect of glibenclamide, an ATP-sensitive K+ (K(ATP)) channels antagonist, on the potassium chloride (KCl)-induced hydroxyl free radical (.OH) generation. Sodium salicylate in Ringer's solution (0.5 nmol/microl per min) was infused directly through a microdialysis probe to detect the generation of .OH as reflected by the formation of dihydroxybenzoic acid (DHBA) in the myocardium of anesthetized rat. The high concentration of KCl (70 mM) significantly increased the level of 2,3- and 2,5-DHBA by the action of depolarization by KCl. However, in the presence of glibenclamide (10 microM), KCl failed to increase the 2,3- and 2,5-DHBA formation. Moreover, when allopurinol (10 mg/kg), a xanthine oxidase inhibitor, was administered by i.v. injection, the elevation of DHBA was not observed. These results suggest that openings of cardiac K(ATP) channel by depolarization evokes .OH generation via xanthine oxidase reaction. PMID- 9857966 TI - Lewy body diseases and multiple system atrophy as alpha-synucleinopathies. PMID- 9857967 TI - Chandelier cells: shedding light on altered cortical circuitry in schizophrenia. PMID- 9857968 TI - Stress and long-term synaptic depression. PMID- 9857969 TI - Promoter prognostication: the serotonin transporter gene and antidepressant response. PMID- 9857970 TI - Leptin: the designer hormone of the 90s. PMID- 9857971 TI - Is the cPLA2 gene associated with schizophrenia? PMID- 9857972 TI - Inositol monophosphatase in immortalized lymphoblastoid cell lines indicates susceptibility to bipolar disorder and response to lithium therapy. PMID- 9857973 TI - Applications of DNA chips for genomic analysis. AB - A major frontier in medical genetics is the definition of the molecular basis of multifactorial diseases. This is especially relevant in the field of clinical psychiatry where the majority of common disorders display complex inheritance patterns, and are further influenced by environmental interactions. New technologies are needed to help address the pressing needs for discovering and deciphering the nature of such disease-associated genes. One such technology which has emerged within the past 3 years involves hybridization-based nucleic acid array (DNA chip) analysis. This technology has the potential to have a lasting impact on diverse genomic-based applications such as large-scale gene mapping studies, mutational analysis, and global expression level monitoring of all human genes. In this review we will describe the fundamental principles behind nucleic acid array-based assays, while focusing on their applications towards genome-wide DNA and RNA analysis. The current capabilities and limitations of these technologies will be discussed, with a focus on areas where future development will be needed for DNA chip-based assays to achieve their full potential. PMID- 9857974 TI - Alpha synuclein is present in Lewy bodies in sporadic Parkinson's disease. AB - A missense mutation in the human alpha synuclein gene was recently identified in some cases of familial Parkinson's disease (FPD). We have developed an antibody that recognizes the C-terminal 12 amino acids of the human alpha synuclein protein and have demonstrated that alpha synuclein is an abundant component of the Lewy bodies found within the degenerating neurons of patients with Parkinson's disease (PD). The presence of alpha synuclein in Lewy bodies of sporadic PD patients suggests a central role for alpha synuclein in the pathogenesis of PD. PMID- 9857975 TI - Beta-amyloid augments platelet aggregation: reduced activity of familial angiopathy-associated mutants. AB - The beta-amyloid (A beta) peptide is present both in serum and in platelets, however it is unclear whether A beta plays a role in platelet function. We have now investigated the effects of soluble A beta on platelet function and have found that low levels (0.1-1 nM) of soluble A beta augment ADP-dependent platelet aggregation and translocation of focal adhesion kinase to the platelet cytoskeleton. Addition of A beta to gel-filtered platelets along with concentrations of adenosine diphosphate (ADP) producing submaximal aggregation responses increased the aggregation response by over 2-fold depending on the ADP:A beta ratios. The structure activity requirements for A beta activity showed intriguing constraints. Only full length A beta has significant activity. Truncated A beta peptides, such as A beta(1-16) or A beta(25-35), or reverse A beta(40-1) all show little or no activity. We also examined the activity of mutant A beta peptides, corresponding with the APP(692A-G) and APP(693E-Q) (at A beta21 and A beta22, respectively) which are found in familial Alzheimer's disease and hereditary cerebral hemorrhagic amyloidosis, Dutch type (HCHWA-D), and found that these peptides showed little or no activity. These results suggest that A beta interacts with platelets in a highly specific manner and may play a role in regulating platelet function. PMID- 9857976 TI - Polymorphism within the promoter of the serotonin transporter gene and antidepressant efficacy of fluvoxamine. AB - Depression with psychotic features has been shown to respond to selective serotonin reuptake inhibitors (SSRIs). The serotonin transporter (5-HTT) is a prime target for SSRIs. A functional polymorphism within the promoter region of the 5-HTT gene, leading to different transcriptional efficiency, was recently reported. We tested the hypothesis that allelic variation of the 5-HTT promoter could be related to the antidepressant response to fluvoxamine and/or augmentation with pindolol (a serotonin autoreceptors antagonist) which has been suggested as an augmentation therapy for nonresponders. One hundred and two inpatients with major depression with psychotic features were randomly assigned to treatment with a fixed dose of fluvoxamine and either placebo or pindolol for 6 weeks. Depression severity was assessed once a week using the Hamilton Depression Rating Scale. Allelic variation in each subject was determined using a PCR-based method. Data were analyzed with a three-way repeated measures analysis of variance. Both homozygotes for the long variant (l/l) of the 5-HTT promoter and heterozygotes (l/s) showed a better response to fluvoxamine than homozygotes for the short variant (s/s). In the group treated with fluvoxamine plus pindolol all the genotypes acted like l/l treated with fluvoxamine alone. Fluvoxamine efficacy in delusional depression seems to be related to allelic variation within the promoter of the 5-HTT gene. Even though other factors may be implicated, genotyping at 5-HTT promoter may represent a promising tool to individualize the pharmacological treatment of depression. PMID- 9857977 TI - Abnormal G protein alpha(s) - and alpha(i2)-subunit mRNA expression in bipolar affective disorder. AB - Disturbances of events associated with intracellular signaling pathways have been suspected of involvement in the development or progression of affective disorders. Often, heterotrimeric G proteins are located at the beginning of these pathways as modulators of extracellular messages. For this reason, messenger RNA expression of three G protein alpha-subunits and of phosphatidylinositol-3 kinase (PI-3 K) regulatory subunit p85 was examined in granulocytes from patients with bipolar or unipolar affective disorder and compared to healthy controls. Messenger RNA expression of the G protein subunit alpha(q) and of p85 was identical in unipolar and bipolar patients and in controls. Furthermore, mRNAs of G protein subunits alpha(s) and alpha(i2) were not different in unipolar patients as compared to healthy controls. Alpha(s) mRNA, however, was markedly increased in bipolar patients. This increase was observed in lithium-treated (more than 12 months) and in unmedicated patients. Elevated levels of alpha(i2) mRNA in unmedicated bipolar patients did not reach statistical significance, whereas mRNA in bipolar patients receiving lithium was significantly above controls. Finally, long-term medication of unipolar patients with lithium had no influence on alpha(i2) mRNA levels. The data reveal elevated mRNA levels of G alpha(s) as a robust feature of bipolar affective disorder. Moreover, despite responsiveness of alpha(i2) gene expression to cAMP-related events, no substantial upregulation of alpha(i2) mRNA was observed in bipolar patients. The lack of alpha(i2) mRNA upregulation, hence, could be an additional abnormality in these patients. Even though lithium was able to reinstate this upregulation, there was no feedback downregulation of alpha(s). This strongly supports the notion of major disturbances of the cAMP signaling system in bipolar illness. PMID- 9857978 TI - Evidence for a chromosome 2p13-14 schizophrenia susceptibility locus in families from Palau, Micronesia. AB - A large multiplex schizophrenia pedigree ascertained from the Micronesian nation of Palau was genotyped with 406 microsatellite DNA markers evenly distributed throughout the genome. Assuming autosomal dominant inheritance, the highest genome-wide lod scores were found for DNA loci mapping to 2p13-14; the maximum lod score was 2.17 (theta = 0.05) at D2S441. A nonparametric APM analysis was also suggestive at D2S441 (APM score = 2.96, P = 0.011). Of the 14 affected cases in this extended family, eight share a large haplotype in this region spanning approximately 11 cM. When 16 other families containing 65 schizophrenic cases were typed in a follow-up study of this region, the maximum lod score remained positive (maximum at D2S441 1.69, theta = 0.20). APM results also remained positive at D2S441 for all 17 families (APM score = 4.87, P = 0.0006). The linkage and haplotype sharing results provide suggestive evidence for a 2p locus predisposing to schizophrenia in a subset of families in the Palauan population. PMID- 9857979 TI - Direct determination of dopamine D4 receptors in normal and schizophrenic postmortem brain tissue: a [3H]NGD-94-1 study. AB - Using an indirect subtraction binding technique and human postmortem tissue, several laboratories reported finding increases in dopamine D4 receptors in caudate nuclei of schizophrenic patients, although others have not replicated these findings. NGD-94-1 is a selective D4 antagonist with low affinity for the D2 and D3 receptors. [3H]NGD-94-1 has been used in this study to directly determine the density of D4 receptors in normals (n = 13) and schizophrenic subjects (n = 7) off antipsychotic drugs for at least 3 months prior to death, or on antipsychotic (n = 7) drugs at the time of death. Human postmortem coronal brain sections were incubated with [3H]NGD-94-1 and autoradiograms developed; and binding in pertinent regions was quantified. In normals, the highest density of [3H]NGD-94-1 binding was in the hippocampus (68 fmol mg(-1), temporal (33), insular (30), and entorhinal cortices (24.9). Significant increases in [3H]NGD-94 1 density in schizophrenics (n = 14) vs normals (n = 13) were observed in the entorhinal cortex (46%) at both low and high magnifications. The increases observed in the schizophrenics were found in both schizophrenics off antipsychotic drugs for at least 3 months prior to death and those on antipsychotic drugs at the time of death. Thus, the changes may be disease related and not a consequence of pharmacological treatment. No significant differences were found between the two schizophrenic groups in any brain area studied. PMID- 9857980 TI - Evidence for a role of phospholipase C-gamma1 in the pathogenesis of bipolar disorder. AB - Several studies have indicated that patients with bipolar disorder (BD) who respond well to lithium prophylaxis constitute a biologically distinct subgroup. Lithium is thought to stabilize mood by acting at the phosphoinositide cycle. We have investigated a polymorphism located in the gene (PLCG1) that codes for a gamma-1 isozyme of phospholipase (PLC), an enzyme that plays an important role in the phosphoinositide second messenger system. A population-based association study and a family-based linkage study were carried out on patients who were considered excellent responders to lithium prophylaxis. Response to lithium was evaluated prospectively with an average follow-up of 14.4 +/- 6.8 years. The PLCG1 polymorphism was investigated in 136 excellent lithium responders and 163 controls. In addition, the segregation of this marker was studied in 32 families ascertained through lithium-responsive bipolar probands. The allele distributions between lithium-responsive bipolar patients and controls were different, with a higher frequency of one of the PLCG1 polymorphisms in patients (chi2 = 8.09; empirical P = 0.033). This polymorphism, however, confers only a small risk (OR = 1.88, CI 1.19-3.00). Linkage studies with the same marker yielded modest support for the involvement of this gene in the pathogenesis of BD when unilineal families were considered (Max LOD = 1.45; empirical P = 0.004), but not in the whole sample. Our results provide preliminary evidence that a PLC isozyme may confer susceptibility to bipolar disorder, probably accounting for a fraction of the total genetic variance. Whether this polymorphism is implicated in the pathogenesis of BD or in the mechanism of lithium response remains to be determined. PMID- 9857981 TI - No evidence for involvement of the leptin gene in anorexia nervosa, bulimia nervosa, underweight or early onset extreme obesity: identification of two novel mutations in the coding sequence and a novel polymorphism in the leptin gene linked upstream region. AB - Mutations in the leptin gene can result in profound obesity in both rodents and humans. In humans, serum leptin levels correlate with body mass index (BMI: kg m( 2)). However, in patients with anorexia nervosa (AN) leptin levels are lower than in BMI-matched healthy controls. We had previously argued that genes involved in weight regulation should be considered as candidate genes for AN. To investigate this hypothesis we screened the coding region of the leptin gene and part of the leptin gene linked upstream region (LEGLUR) in 49 patients with AN and 315 children and adolescents with extreme obesity. Two novel mutations in the coding region (Ser-91-Ser; Glu-126-Gln), each found in a single proband, and a novel polymorphism in the LEGLUR (position -1387 G/A; frequency of both alleles approximately 0.50) were identified. Tests for association of LEGLUR polymorphism alleles were negative by comparing allele frequencies between 115 AN patients, 71 bulimia nervosa patients, 315 extremely obese children and adolescents, 141 healthy underweights and 50 controls that were not selected for body weight. Tests for transmission disequilibrium were also negative. Hence, an influence of variations in the leptin gene on eating disorders or extreme early onset obesity could not be detected. PMID- 9857982 TI - Leptin in relation to resumption of menses in women with anorexia nervosa. AB - Serum levels of leptin are decreased in underweight AN patients and increase with weight restoration. To assess the relationship of decreased leptin levels with other hormonal abnormalities in AN and to evaluate the possible role of increasing leptin levels, alone or in combination with other hormones, in the resumption of menses that accompanies weight gain, we studied cross-sectionally sixty-five consecutively enrolled AN patients. Subjects were divided in three groups: (I) underweight and amenorrheic; (II) weight-recovered but still amenorrheic; and (III) weight-recovered and eumenorrheic women. Patients in group I had decreased BMI, serum leptin, estradiol (E2), insulin-like growth factor 1 (IGF-1) and urinary growth hormone (GH) levels and increased sex hormone-binding globulin (SHBG) levels, compared to AN patients in groups II and III. Moreover, although no differences in leptin levels or BMI were observed between amenorrheic and eumenorrheic weight-recovered patients (groups II and III), free E2 and GH levels were higher (P<0.02) in weight-recovered, eumenorrheic women. Thus, it appears that leptin is a necessary, but not a sufficient, factor for the resumption of menses in AN patients. PMID- 9857984 TI - Nucleotide sequence and phylogenetic classification of human papillomavirus type 67. AB - The complete nucleotide sequence of human papillomavirus type 67 (HPV 67) cloned from a vaginal intraepithelial neoplasia, has been determined. It consists of 7801 nucleotides with a GC content of 38.4% and exhibits similar genome organizations of genital HPVs. By phylogenetic analysis based on the full nucleotide sequences of E6 open reading frame of 28 genital HPVs, HPV 67 was clustered with HPV 16, 31, 33, 34, 35, 52, and 58. PMID- 9857983 TI - Orf virus encodes a homolog of the vaccinia virus interferon-resistance gene E3L. AB - A homolog of the vaccinia virus (VAC) interferon resistance gene E3L has been discovered in orf virus strain NZ-2, a parapoxvirus that infects sheep, goats and humans. The gene is located 20 kb from the left terminus of the orf virus genome and is transcribed towards this terminus. RNase protection studies have been used to define the limits of the gene and Northern analysis revealed that it is expressed early in infection. The predicted amino acid sequence of the orf virus protein shares 31% identity (57% similarity) with the VAC E3L protein. Four of the six residues identified as being essential to dsRNA binding in the vaccinia virus protein are conserved in the orf virus protein whilst the other two amino acid changes are conservative substitutions. The orf virus gene has been sequenced in two other orf virus strains which vary markedly in their ability to produce experimental lesions in vivo. Their predicted protein sequences vary by less than 3% from the NZ-2 protein. The recombinant orf virus protein, expressed as a fusion protein in E. coli, bound double-stranded (ds)RNA but not dsDNA, single-stranded (ss)DNA or ssRNA . This is the first demonstration of a VAC E3L like gene encoded by a parapoxvirus. PMID- 9857985 TI - Primary structures of hemagglutinin-esterase and spike glycoproteins of murine coronavirus DVIM. AB - Diarrhea virus of infant mice (DVIM) is a member of murine hepatitis viruses (MHVs). The nucleotide sequences of the genes encoding the hemagglutinin-esterase (HE) and the spike (S) glycoproteins from DVIM were determined and compared with those of other MHVs. The deduced amino acid sequence of the HE protein was most similar to that of MHV-S strain (94% identity), and the S protein sequence was most similar to that of MHV-Y strain (90% identity). The DVIM HE protein has a unique N-linked glycosylation site in addition to other glycosylation sites common to many MHV strains. Unlike in some typical MHV strain, such as MHV-A59 and MHV-JHM, the vast majority of the S glycoprotein molecules in DVIM exist an uncleaved form probably due to several amino acid substitutions around the cleavage site. PMID- 9857986 TI - Expression analysis of recombinant herpes simplex virus type 1 DNase. AB - Expression of recombinant herpes simplex virus type 1 (HSV-1) deoxyribonuclease (DNase) was analyzed in BHK-21 cells, a standard cell line for virus propagation, by using mammalian cell expression systems based on vaccinia virus and on Semliki Forest virus (SFV)1. Although the establishing of recombinant vaccinia virus failed due to the apparent toxicity of the herpesviral enzyme, soluble and functional HSV-1 DNase was efficiently expressed in BHK-21 cells by the vaccinia virus/T7 RNA polymerase hybrid system as well as by recombinant Semliki Forest virus. Using rabbit antiserum ExoC, directed against the C-terminal residues 503 626, or mouse monoclonal antibody (MAb) Q1, raised against the type 2 enzyme, a major 85-kDa protein with the identical size of the enzyme from HSV-1-infected cells was identified to be induced in both expression systems. With recombinant SFV functional HSV-1 DNase coincided with the overproduction of a single major 85 kDa protein reaching an optimum between 16 h and 36 h after infection. At later times of infection the enzymatic activity vanished. Thus, recombinant SFV may be an appropriate expression vector for biochemical studies of the enzyme when (i) packaged recombinant virus particles are used for infection and (ii) infection does not exceed 24 h. Due to the limitations of transient expression systems, the vaccinia/T7 RNA polymerase hybrid system is suited for expression analysis on a small scale, and for studying intracellular interactions of the enzyme as demonstrated by immunofluorescence microscopy studies. Using vector pTM1, recombinant HSV-1 DNase was efficiently overproduced in BHK-21 cells at 6 h after transfection and was shown to colocalize with the cellular chromatin at sites apparently distinct from the bulk of the herpesviral replication sites the way it is observed for the enzyme of lytically infected cells. The deleting of the 123 C terminal amino acid residues did not alter this nuclear localization of HSV-1 DNase, suggesting that the latter sequences and other herpesviral factors are not required for the chromatin association. PMID- 9857987 TI - The cauliflower mosaic virus capsid protein: assembly and nucleic acid binding in vitro. AB - The capsid protein of the cauliflower mosaic virus (CaMV) was expressed in a bacterial system to study CaMV assembly. Bacterial lysates contained soluble particulate material and insoluble inclusion bodies that were both used for analysis. In vitro renaturation of pIV derivatives lead to the appearance of folded sheets or large tubular structures in electron microscopy. The region between amino acid positions 77 and 332 is sufficient for self-aggregation of pIV in vitro. C-terminal deletion to amino acid position 265 still allowed dimerization but prevented further aggregation. Nucleic acid binding assays of immobilized pIV derivatives demonstrated that a region located upstream of the retroviral "zinc finger-like" motif is involved in unspecific binding dsDNA, ssDNA and RNA. PMID- 9857988 TI - Sequence comparison of the large genomic RNA segments of two strains of lymphocytic choriomeningitis virus differing in pathogenic potential for guinea pigs. AB - Two strains of lymphocytic choriomeningitis virus (LCMV) differ in their ability to cause a lethal disease in outbred guinea pigs: the Armstrong (ARM) strain is not lethal at high doses (10(6) PFU), whereas the WE strain is lethal at less than 10 PFU inoculated intraperitoneally. The high pathogenic potential of LCMV WE has been mapped to the larger (L) of the two genomic RNA segments by genetic reassortment analysis (Riviere, Y., Ahmed, R., Southern, P. J., Buchmeier, M. J. and Oldstone, M. B. A., J. Virol. 55, 704-709, 1985). Here we describe the completed sequence of the LCMV WE L RNA, and its comparison to the L RNA of the non-virulent strain, LCMV ARM. Similar to the L RNA of LCMV ARM, the L RNA of WE is 7.2 kb long and contains two open reading frames (ORFs): the 5" ORF encodes a small RING finger (zinc-binding) protein, p11 Z, and the 3" ORF encodes the putative RNA-dependent RNA polymerase (RdRp or L protein). Comparison of nucleotide sequences for both viruses revealed 84% L RNA homology. At the amino acid level similarity between the two strains is 87% in the Z ORF, and 88% in the RdRp ORF. The most divergent regions are found in the N-terminal parts of the RdRp and Z proteins and are most likely to account for differences in pathogenic potential. PMID- 9857990 TI - Sequence characteristics of natural populations of tomato spotted wilt tospovirus infecting flue-cured tobacco in Georgia. AB - Using primers from conserved regions, the nucleocapsid (Nc) gene sequences of naturally occurring tomato spotted wilt tospovirus (TSWV) isolates from flue cured tobacco (Nicotiana tabacum) grown in several Georgia counties were amplified by immunocapture-reverse transcription-polymerase chain reaction. The resulting amplicons were cloned and sequenced. Sequence analyses showed a high degree of sequence conservation among the Nc genes of the tobacco isolates, and those reported from other parts of the world. However, distinct differences that were unique to these tobacco isolates as well as the previously studied peanut, tomato and pepper isolates from Georgia were present. The Georgia isolates formed a distinct cluster that was clearly resolved from the rest of the TSWV isolates based on sequence phylograms. PMID- 9857989 TI - Envelope and long terminal repeat sequences of an infectious murine leukemia virus from a human SCLC cell line: implications for gene transfer. AB - Development of methods for gene transfer into specific cell types or tissues is important for experimental research as well as clinical therapeutical approaches. We report here the cloning and characterization of the envelope (env) gene and the U3 region of a retrovirus from an infected human Small Cell Lung Cancer (SCLC) cell line. The replication of this murine retrovirus is also fully supported by other lung cancer cell lines of different histological origin. We present evidence that a long terminal repeat (LTR)-beta-galactosidase (beta-Gal) reporter construct performed as well as an analogous cytomegalovirus (CMV) promoter beta-Gal construct in the human lung epithelial cell line A549 and in the human larynx carcinoma cell line HEp2. PMID- 9857991 TI - A novel spliced transcript of influenza A/WSN/33 virus. AB - It has been shown that influenza A virus M1 mRNA has two alternative 5" splice sites: a distal 5" splice site producing M3 mRNA that has the coding potential for 9 amino acids and a proximal 5" splice site producing M2 mRNA that encodes the essential M2 ion-channel protein. In this study, we demonstrated that the laboratory widely used strain A/WSN/33, but not A/Udorn/72 possessed another novel 5" splice site producing a transcript with the coding potential for 54 amino acids. We nominated this novel transcript as M4 mRNA. M4 mRNA was detected in A/WSN/33-infected cells derived from different species. Sequence comparison of M1 mRNA in both A/WSN/33 and Udorn/72 at position 143 to 151 reveals that this novel 5" splice site generated in WSN was due to one nucleotide difference at position 147. Several strains of influenza A viruses other than WSN also possess the potential M4 5" splice site by sequence analysis from the files of GenBank. PMID- 9857992 TI - Stable expression of nucleocapsid proteins of Puumala and Hantaan virus in mammalian cells. AB - The development of an in vitro-system for the stable expression and the analysis of native hantavirus proteins using hantaviral cDNA is of particular interest. As a first step the expression of the hantavirus nucleocapsid (N) proteins in mammalian cells was studied in more detail. The cDNA of the S-RNA segment of Puumala virus strain CG-1820 and Hantaan virus strain 76-118 was used for the construction of eucaryotic expression vectors that allow the generation and selection of mammalian cells harboring and expressing the N protein genes of hantaviruses. A variety of conventional and novel expression vectors as well as different mammalian cell lines were screened. The expression of the N protein of Puumala virus using the pGRE5-1 vector in which the transcription is under control of inducible glucocorticoid responsive elements (GRE) revealed that the Puumala virus N protein can be expressed in Vero E6 cells efficiently without any detectable cell toxicity. From the variety of expression vectors tested, it was found that pCR3.1 is the vector of choice for stable expression of hantavirus N proteins. The successful establishment of different mammalian cell lines expressing considerable amounts of Puumala and Hantaan virus N protein indicates that the stable and efficient expression of this particular viral protein in the cell lines of three evolutionary distinct species (human, monkey, and mouse) is possible. The system described here represents the experimental basis for further studies of hantavirus infection, replication, and pathogenesis using a reverse genetics approach. PMID- 9857993 TI - Lung cancer and smoking: a case-control study in Buenos Aires, Argentina. AB - The objective of this study was to examine the risks for lung cancer associated with lifestyle characteristics of smoking in a developing country where lung cancer is the first cause of mortality by cancer in men, tobacco propaganda is freely allowed, and there are no restrictions operating for smoking. The design was a case-control and hospital-based study. Two hundred men with lung cancer and 397 hospital controls were interviewed. Odds ratio (OR) for current smokers was 8.5, whereas former smokers displayed an OR of 5.3. The risk increased with duration of smoking and with the number of cigarettes smoked per day. The attributable risk for smoking was 85%. Smokers of black tobacco and more than 24 cigarettes/day showed a risk of 12.9 regarding non-smokers, and of 15.5 for 40 or more years duration of smoking. The proportion of cases diagnosed as adenocarcinoma was higher than the proportion of squamous cell carcinoma. PMID- 9857994 TI - Correlation of expression of bombesin-like peptides and receptors with growth inhibition by an anti-bombesin antibody in small-cell lung cancer cell lines. AB - The murine anti-bombesin monoclonal antibody, 2A11, has been demonstrated to inhibit growth of some small-cell lung cancer (SCLC) cells in nude mice xenografts and in a clinical trial. To determine if the expression of bombesin like peptides (BLP) and their receptors (GRP-R and NMB-R) correlate with an in vitro response to 2A11, we measured these parameters in seven SCLC cell lines. Gastrin releasing peptide (GRP) mRNA was detected in three of seven cell lines (NCI-H69, NCI-H345, NCI-H510) and neuromedin B (NMB) mRNA was detected in all seven lines using an RNase protection assay (RPA). Immunoreactive BLP was detected in the cell pellets of all lines (range 0.11-59.90 pmol/mg protein) by a solid phase GRP radioimmunoassay (RIA) using 125I-labeled 2A11. RPA detected GRP receptor mRNA in two cell lines (NCI-H69 and NCI-H345) and NMB-receptor in three lines (NCI-H345, NCI-H510, and NCI-H660). Reverse transcriptase-PCR confirmed the presence of receptor mRNA in these lines and detected NMB-receptor in an additional three lines (NCI-H69, NCI-H82, and NCI-H187). Calcium mobilization in response to BLP stimulation was detected in the six cell lines expressing either GRP-R or NMB-R mRNA but not in NCI-N417, which had no detectable BLP-receptor. 2A11 (5 microg/ml) inhibited colony formation by 26-61% after 2 weeks in all cell lines except NCI-N417. Thus, growth inhibition by 2A11 requires the presence of at least one BLP-receptor. These findings may be useful in selecting patients with SCLC for treatment with 2A11. PMID- 9857995 TI - Selective use of PET scan in the preoperative staging of NSCLC. AB - Positron emission tomography (PET) is a imaging technique which identifies abnormalities by utilising their metabolic properties rather than their anatomical density. In this study, we evaluated the place of PET scanning for the characterisation of additional abnormalities discovered on routine, preoperative CT evaluation of patients with proven NSCLC. A total of 21 patients underwent PET scanning using fluorodeoxyglucose (FDG). In these patients, routine CT scans had found 26 concomitant lesions which were in lung, adrenal gland, liver, kidney, spleen and pleura. FDG uptake was positive at the primary site in all patients. FDG uptake in 13 of the concomitant lesions suggested malignancy. Malignancy was confirmed at all of these foci by histology in eight (six at thoracotomy, fine needle aspiration biopsy (FNAB) and open biopsy) and by clinical and radiological follow-up for 1-15 months in five. In 13 lesions, PET suggested benign disease. Histology reported no evidence of malignancy in five and the six foci followed on clinical and radiographic follow-up for 2-16 months had no evidence of malignancy. PET was falsely negative in only one site where malignancy was proven by thoracotomy. PET failed to detect the remaining lesion which was an unassociated primary kidney tumour in a patient with Li-Fraumeni syndrome. The accuracy of PET scanning in the assessment of those additional lesions found on CT evaluation of otherwise operable NSCLC was found to be 96% with a sensitivity of 93% and specificity of 100%. These results indicate that PET is useful to characterise additional lesions found on CT scan of patients with NSCLC. PMID- 9857996 TI - Peripheral non-small cell lung cancers 2.0 cm or less in diameter: proposed criteria for limited pulmonary resection based upon clinicopathological presentation. AB - Clinical features of peripheral non-small cell lung cancer 2.0 cm or less were retrospectively analyzed. Nodal status and prognosis in relation to tumor diameter and histologic type were investigated in 171 consecutive patients with peripheral clinical T1N0M0 non-small cell lung carcinomas 2 cm or less in diameter and who had undergone surgical resection between 1976 and 1997. Of the 171 patients, 136 had adenocarcinoma, 27 had squamous cell carcinoma, four had large cell carcinoma, three had carcinoid and one had adeno-squamous carcinoma. There was no statistically significant difference in the incidence of stage I cases between adenocarcinoma and squamous cell carcinoma. Lymph node involvement was recognized in 30 (17.5%) patients: ten (5.8%) at N1 nodes and 20 (11.7%) at N2 nodes. Lymph node metastasis was significantly more common in tumors 1.5-2.0 cm in diameter (22%) than in those 1.5 cm or less in diameter (14.0%, P = 0.0490). There was no lymph node metastasis in tumors 1.0 cm or less in diameter. The 5-year survival rates cases with or without lymph node involvement were 63.3 and 75.3%, respectively, showing significant difference (P = 0.0338). The result of the present study suggested that systematic mediastinal and hilar lymph node dissection is necessary even for cases with tumor diameter less than 2 cm. However, if the tumor is within 1.0 cm in diameter, mediastinal lymph node dissection might be dispensable; therefore, these cases are good candidates for video-assisted lobectomy. PMID- 9857997 TI - Controlled clinical trial of prophylactic cranial irradiation for patients with small-cell lung cancer in complete remission. AB - We conducted a randomised clinical trial on 211 patients with small-cell lung cancer in complete remission (CR). The aim of this trial was to evaluate the effect of prophylactic cranial irradiation (PCI) on overall survival. Eligible patients were randomly assigned to receive either PCI (100 patients) or no PCI (111 patients). Each centre was allowed to use its own PCI protocol as long as the total dose was within the range of 24-30 Gy and delivered in less than 3 weeks with fractions of 3 Gy or less. The mean follow-up is 5 years. The survival curves do not differ significantly (P = 0.25) between the two groups. The 4-year overall survival rate (95% confidence interval) is 22% [15-32%] in the PCI group versus 16% [10-25%] in the control group. The relative risk of death in the PCI group compared to the control group is 0.84 (95% CI = [0.62-1.13]). The incidence of brain metastasis is lower in the PCI group, but the difference is not statistically significant (P = 0.14). The 4-year cumulative rate of brain metastasis is 44% [32-57%] in the PCI group compared to 51% [38-63%] in the control group. In conclusion, in this study, which had to be closed prematurely, no significant difference was found in terms of the incidence of brain metastases nor in survival. PMID- 9857998 TI - Analysis of neuroendocrine markers, HER2 and CEA before and after chemotherapy in patients with stage IIIA non-small cell lung cancer: a Cancer and Leukemia Group B study. AB - Several studies have suggested that biochemical or molecular markers examined in non-small cell lung cancer carry prognostic or treatment response information. Non-small cell lung cancer patients whose tumors have neuroendocrine (NE) features may be more responsive to chemotherapy. In addition, increased expression of HER2 (c-erbB-2), a membrane-bound receptor with tyrosine kinase activity, has been associated with shortened survival. The Cancer and Leukemia Group B (CALGB) performed a study of patients with stage IIIA (N2 nodes positive) non-small cell lung cancer in which patients received initial chemotherapy followed by surgery, then post-operative therapy consisting of sequential chemotherapy and radiation therapy. Since all patients underwent mediastinoscopy, this provided an opportunity to compare pre- and post-chemotherapy tumor specimens to test the hypothesis that these proteins would predict treatment response. In particular, we hypothesized that the post-chemotherapy specimens would be enriched for NE marker negative cells because of the increased sensitivity of NE positive cells to chemotherapy. We performed immunohistochemical analysis for a panel of NE markers [neuron-specific enolase (NSE), Leu-7, chromogranin A (ChrA), synaptophysin (Syn)], HER2 and CEA to determine if there was an effect of therapy on the percentage of cells expressing these markers. Secondary endpoints were a correlation with chemotherapy response and survival. Slides were scored for intensity (0-4) and percentage of cells positive (0-4). Of 61 eligible patients, there were 38 with both pre- and post chemotherapy specimens. When both intensity of staining and percentage of positive cells were considered, post-chemotherapy specimens had a higher percentage of positive NE markers compared with pre-chemotherapy. In addition, there was no correlation between NE marker, HER2 or CEA expression (prior to or post treatment) and response to chemotherapy or survival. These data do not support the hypothesis that NE positive tumor cells are preferentially killed by chemotherapy in patients with stage IIIA non-small cell lung cancer. PMID- 9858000 TI - Maintenance chemotherapy for small cell lung cancer. PMID- 9857999 TI - Combination chemotherapy with docetaxel, vinorelbine and cisplatin as first-line treatment of advanced non-small-cell lung cancer: a multicenter phase II study of the Greek Cooperative Group for Lung Cancer. AB - Vinorelbine, docetaxel and cisplatin have documented single-agent activity in non small-cell lung cancer (NSCLC); a multicenter phase II trial was initiated in order to evaluate the tolerance and efficacy of their combination. A total of 24 chemotherapy-naive patients with measurable stage IIIB or IV NSCLC and performance status (PS; WHO) 0-2 entered the study. Vinorelbine (20 mg/m2 i.v.) was given on days 1 and 15, cisplatin (60 mg/m2) on day 1, and docetaxel (100 mg/m2) on day 16, in cycles of 28 days. Recombinant human granulocyte colony stimulating factor (150 microg/m2 s.c.) was administered prophylactically from day 17 to day 27. One pathological complete (4%) and six partial responses (25%) were documented (overall response 29%; 95% CI 11.6-49.2%). A total of five patients (21%) had stable and 12 (50%) progressive disease. The median duration of response was 28 weeks and the median time to tumor progression 36 weeks; the median survival was 20 weeks. Grade 3-4 neutropenia occurred in 16 patients (67%) while 13 of them (54%) developed febrile neutropenia. Grade 4 mucositis occurred in two patients (8%) and one of them also presented grade 4 diarrhea. There were four treatment-related deaths: two from sepsis, one from massive hemoptysis due to a pulmonary abscess and one from acute myocardial ischemia 7 days post chemotherapy. In conclusion, the high incidence of neutropenic episodes and treatment-related deaths led to an early discontinuation of patient enrollment. This combination, in the schedule and the doses used, could not be recommended for off protocol treatment of patients with advanced NSCLC. PMID- 9858001 TI - Retinal arteriovenous malformation with fluctuating vision and ischemic central retinal vein occlusion and its sequelae: 25-year follow-up of a case. AB - A patient with a retinal arteriovenous malformation (AVM) had experienced episodic visual loss with spontaneous recovery for many years, followed by permanent visual loss secondary to central retinal vein occlusion. She subsequently progressed to development of retinal neovascularization extending onto the posterior vitreous face with recurrent vitreous hemorrhage requiring vitrectomy. A brief review of the literature is presented, and the cause of the fluctuations in vision and central retinal vein occlusion are discussed. PMID- 9858002 TI - Retinal ischemia in aortic arch atheromatous disease. AB - Retinal ischemia is often caused by emboli arising from the cardiac chambers or the common carotid artery bifurcation; the latter are often composed of cholesterol. However, in many patients no lesions are identified after evaluation of these sources of emboli. Two patients were observed who had retinal ischemia and emboli originating from aortic atheromatous plaques that were visualized by transesophageal echocardiography. Cardiac, carotid, and intracranial sources of emboli were excluded. The embolic nature of retinal ischemia was further corroborated by the presence of microembolic signals during transcranial Doppler insonation of the middle cerebral artery on the side ipsilateral to the symptomatic retina. In patients with Hollenhorst plaques the aortic arch can be a potential source of emboli. Transesophageal echocardiography should be considered in these patients when the initial evaluation does not identify a cardiac or carotid lesion. PMID- 9858003 TI - Monocular band optic atrophy. AB - Band or "bow tie" optic atrophy is characterized by well-described ophthalmoscopic findings in the optic nerve and nerve fiber layer and homonymous hemianopia. It is typically associated with compressive lesions of the pregeniculate postchiasmal visual pathway or, less commonly, congenital malformations affecting the postgeniculate radiations or cortex. A unique case with band optic atrophy is described because of the unilateral visual defect. The optic atrophy is strictly unilateral and without an obvious structural lesion that could explain the optic disc damage. However, incidental cerebral gray matter cortical heterotopia may mark a congenital insult that contributed to both of these abnormal findings. PMID- 9858004 TI - Unusual presentations of sellar arachnoid cyst. AB - This report describes two unusual cases of parasellar arachnoid cyst with different neuro-ophthalmologic manifestations and clinical courses: a 33-year-old woman with parasellar arachnoid cyst, manifested by incongruous homonymous hemianopia, and a 64-year-old man with a presumed parasellar arachnoid cyst and bitemporal hemianopia that subsequently decompressed spontaneously. Parasellar arachnoid cyst is uncommon, and the clinical course has been incompletely described in the literature. Optimal treatment of patients with these cysts necessitates better understanding of their signs, symptoms, and clinical course. PMID- 9858005 TI - Homonymous hemianopsia due to a dural cavernous hemangioma. AB - The clinical and radiographic features of extra-axial cavernous hemangiomas are described, and a case of homonymous visual field loss due to a dural-based occipital cavernous hemangioma is reported. A patient presented with a homonymous hemianopsia due to an enhancing tentorial mass lesion. The preoperative clinical and magnetic resonance imaging features were suggestive of meningioma. The patient underwent gross total resection of the lesion and the final pathologic examination was consistent with cavernous hemangioma. There was complete resolution of the visual field defect after surgery. Extra-axial cavernous hemangiomas differ from intra-axial cavernous hemangiomas in their clinical and radiographic features. The former lesions may mimic meningioma and should be considered in the differential diagnosis of a dural-based mass. Early recognition of the lesion is important because surgical removal of cavernous hemangiomas may be associated with a higher morbidity and mortality rate than meningiomas. PMID- 9858006 TI - When fighting makes you see black holes instead of stars. AB - A young boxer developed a left homonymous hemianopia immediately after a blow to the head. A magnetic resonance image showed a lesion in the right lateral geniculata region that was consistent with a previous cerebral hemorrhage and was likely the cause of the visual field defect. PMID- 9858007 TI - Variability in visual cortex activation during prolonged functional magnetic resonance imaging. AB - This study was conducted to test whether cortical activation varies across successive epoques during functional magnetic resonance imaging (fMRI) studies. Ten normal adult volunteers were studied with a 1.5-T MR scanner. Pseudocoronal study planes were chosen perpendicular to the tentorium cerebelli, at two thirds the distance from the posterior edge of the splenium of the corpus callosum to the transverse sinuses. Functional images were acquired with a T2*-weighted spoiled gradient echo sequence. The visual cortex was stimulated by goggles flashing at 8 Hz. Each study consisted of 82 sequential scans, lasting 15 seconds each for a total of 20.5 minutes. Two scans without stimulation were alternated with two scans of visual stimulation. Scans 3 through 83 were divided into five sequences of 16 scans. For each sequence, the number of pixels within a predefined rectangular region of interest that showed increased activity during stimulation were counted. Least squares regression models of straight lines were fit to the data. The initial level of visual cortex activation in the region of interest, as measured by the y-intercept, varied substantially from subject to subject (range: 4-68, p < 0.001). There was sufficient evidence of systematic change with time to reject the hypothesis of constant activation with the same stimulus over time (p=0.02). The observed visual cortex activation with single plane fMRI varied both with time over successive epoques and among subjects. Possible factors responsible for the variation may include head movement, eyelid position, attention, and physiologic fatigue. These factors must be accounted for in experimental design and in data analysis and interpretation. PMID- 9858008 TI - Visually induced reactivity in posterior cerebral artery blood flow. AB - To evaluate visually induced reactivity (VIR) in the posterior cerebral artery (PCA), mean flow velocities in the PCA were measured bilaterally in 35 normal subjects and in 17 patients with PCA territory infarctions, by means of transcranial Doppler ultrasound. After the individual PCA baseline flow was estimated, different visual stimuli were applied: on-off light, colored light, complex scene, and visual imagery task, and the CO2 test was administered. A sampling rate of 20 Hz was used, and the raw data were transferred to a computer. The baseline flow and the maximum flow increase were calculated with a specially designed program. In control subjects, the on-off light stimulus induced a mean increase in PCA flow velocities of 21.5+/-6.4%, and colored light induced an increase of 22.3+/-6.3%. Complex scenes significantly elevated VIR more than light and colored light, with a mean increase of 28.8+/-6.8% (p < 0.05). Mental imagery had no significant effect on PCA flow velocities. There was no significant difference in flow between the right and left PCA in healthy subjects. In patients with PCA territory infarctions with homonymous hemianopsia or quadrantanopsia, there was a marked decrease of VIR and CO2 reactivity on the affected side corresponding to the extent of PCA territory infarction. Visual stimuli increased blood flow velocity bilaterally in the PCA, which supply the visual cortex and visual association area. This noninvasive test seems to be well suited to normal subjects and to patients with vascular disorders affecting the PCA. PMID- 9858009 TI - The many faces of sarcoidosis. PMID- 9858010 TI - Sinus histiocytosis with massive lymphadenopathy involving the orbit: reversal of compressive optic neuropathy after chemotherapy. AB - A 38-year-old woman from Antigua had compressive optic neuropathy of the right eye caused by orbital involvement with sinus histiocytosis. There was also nasal sinus involvement and massive cervical lymphadenopathy resulting in radiographic compression of the airway and carotid sheath. Because of the compressive optic neuropathy and threat to the airway and carotid perfusion, the patient underwent a 6-month chemotherapeutic regimen of cyclophosphamide, vincristine, and prednisone. After chemotherapy, the visual dysfunction resolved in correlation with diminution of the orbital mass, and marked regression of the cervical lymphadenopathy. This case demonstrates the potential efficacy of chemotherapy in the treatment of compressive optic neuropathy in cases of orbital sinus histiocytosis with massive lymphadenopathy. PMID- 9858011 TI - Tamoxifen retinopathy. PMID- 9858012 TI - Bilateral internuclear ophthalmoplegia related to chronic toluene abuse. AB - A 36-year-old woman exhibited slurred speech, progressive ataxia, blurred vision, and oscillopsia. Examination showed dysconjugate torsional nystagmus and bilateral internuclear ophthalmoplegia (INO). Further investigation revealed evidence of chronic toluene abuse. The neurologic findings in toluene abuse and the causes of bilateral INO are discussed. PMID- 9858013 TI - Mobius syndrome with oculomotor nerve paralysis without abducens paralysis. AB - Mobius syndrome is typified by bilateral facial nerve palsies, usually with abducens palsies. We examined an infant with Mobius syndrome who had bifacial weakness and third nerve palsies, but intact abduction of both eyes. Lower cranial nerve involvement, leading to respiratory, swallowing, and cardiac difficulties, was also present. Pathologic examination of the brainstem showed absent or hypoplastic third, seventh, tenth, and twelfth nerve nuclei. The fourth, fifth, sixth, and eighth nerve nuclei were intact. In Mobius syndrome with ocular motor palsies, rarely the sixth nerve may be spared. PMID- 9858014 TI - Patient with Kearns-Sayre syndrome exhibiting abnormal magnetic resonance image of the brain. AB - A 33-year-old Japanese man had Kearns-Sayre syndrome (KSS), which consists of the triad of external ophthalmoplegia, heart block, and "salt-and-pepper" retinopathy. The other systemic manifestations included sensorineural hearing loss, slight generalized muscle weakness, cerebellar ataxia, and elevated levels of cerebrospinal fluid protein. He exhibited a heteroplasmic mitochondrial DNA deletion of approximately 9 kb between the cytochrome c oxidase subunit 1 and cytochrome b genes. In the authors' experience, this deletion is one of the longest to be observed in such patients. His fundi were characterized bilaterally by white flecks in the inner layers of retina at the midperiphery. Visual evoked potentials showed delayed latency in the P100 component. The tibial somatosensory evoked potential revealed a marked prolongation of interpeak latency between the N20 and P40 components. Brain magnetic resonance images revealed high-intensity foci in several regions on T2-weighted images. Electrophysiological and magnetic resonance imaging findings suggested an involvement of the white matter of the central nervous system in this patient that was not reflected in the clinical findings. PMID- 9858015 TI - Proptosis with acute oculomotor and abducens nerve palsies. AB - OBJECTIVE: To determine whether orbital axial proptosis occurs in the affected eye of patients with acute oculomotor or abducens nerve palsy. MATERIALS AND METHODS: In this prospective, cross-sectional survey, the Hertel instrument was used to measure the amount of axial protrusion in 26 consecutive patients with complete or severe acute oculomotor nerve palsy and 27 consecutive patients with complete or severe acute abducens nerve palsy. The Mann-Whitney test was used to compare the amount of relative proptosis of the affected eye in the patient groups with that of 40 control subjects. The absolute amount of the interocular difference in axial protrusion of the normal eyes was used to determine control values. The proportion of patients and control subjects with relative proptosis greater than 1 mm was compared using Fisher's exact test. RESULTS: There was no significant difference in the degree of relative proptosis of the affected eye of patients with oculomotor nerve palsy or abducens nerve palsy as compared with the control eyes. Likewise, there was no significant difference in the proportion of patients with relative proptosis greater than 1 mm in either the oculomotor nerve palsy group or the abducens nerve palsy group as compared with the control group. CONCLUSIONS: Acute oculomotor or abducens nerve palsy does not produce any significant amount of orbital axial proptosis in the affected eye, at least as measured clinically using the Hertel instrument. Acute neurogenic ophthalmoplegia rarely causes relative proptosis greater than 1 mm. PMID- 9858016 TI - Epileptic periodic alternating nystagmus. AB - A patient developed periodic alternating nystagmus, periodic alternating gaze deviation, and periodic alternating head rotation as a manifestation of a seizure. This occurred as he awakened after hypoxic ischemic encephalopathy. Seizures should be added to the list of differential diagnoses of periodic alternating nystagmus. PMID- 9858017 TI - Functional magnetic resonance imaging (fMRI) of the occipital cortex in the presence of post-papilloedematous optic atrophy. PMID- 9858018 TI - Mild hypothermia--a revived countermeasure against ischemic neuronal damages. AB - Although hypothermia as a means of cerebral protection against and resuscitation from ischemic damage has a history of approximately six decades, extensive studies, both in basic and clinical fields, on the mechanisms, effects and methods of mild hypothermia at temperatures no less than 31 degrees C have started only in the last decade. In experiments on rodents, hypothermia in the postischemic period that is introduced up to several hours after reperfusion and is maintained for one day followed by a slow rewarming, significantly protects hippocampal neurons against damage. The mode of action of hypothermia is apparently non-specific and multi-focal in widely progressing cascade reactions in ischemic cells; namely, suppressing: (1) glutamate surge followed by; (2) intraneuronal calcium mobilization; (3) sustained activation of glutamate receptors; (4) dysfunction of blood brain barrier; (5) proliferation of microglial cells; and (6) production of superoxide anions and nitric oxide. In addition, mild hypothermia modulates processes in ischemic condition at the level of cell nucleus, such as the binding of transcription factor AP-1 to DNA, and ameliorates the depression of protein synthesis. This non-specific and widely affecting manner might explain why hypothermia is superior to any medicine developed. Recent clinical trials of mild hypothermia in various individual institutions have revealed significantly beneficial outcomes in some cases, along with an accumulation of practical knowledge of techniques and treatments. Large scale randomized studies involving multiple institutions as well as exchange of informations and ideas are needed for further development of hypothermia treatment. PMID- 9858019 TI - Bradykinin and muscle stretch alter contralateral cat neck muscle spindle output. AB - The objective of this study was to investigate the activity of primary and secondary muscle spindle afferents (MSAs) in neck muscles, when the contralateral splenius or trapezius muscles were (1) exposed to i.m. injection of bradykinin (BK) (6-86 microg/ml), (2) stretched, (3) stretched during exposure to BK or (4) stretched after exposure to BK. It was found that injection of BK, muscle stretch and the combination of the two stimuli significantly excited primary and secondary MSAs via reflex effects onto static fusimotor neurones. BK, alone, and in combination with muscle stretch, induced more frequent, potent and long lasting effects as compared to muscle stretch. The effects of muscle stretch was significantly increased after exposure of BK, indicating that stretch-sensitive nerve-endings within neck muscles are sensitised by BK. The results are discussed in relation to the disturbances in motor co-ordination and proprioception found in patients suffering from chronic muscle pain in the cervical region. PMID- 9858020 TI - Upper airway motor outputs during sneezing and coughing in decerebrate cats. AB - The purposes of the present study were to determine which upper airway movements cause a difference in the expiratory airflow pathway between sneezing and coughing, and to develop a new animal model for studying the neural mechanism of sneezing in paralyzed animals, i.e. fictive sneezing. We compared the upper airway motor patterns of sneezing and coughing, induced by electrical stimulation of the anterior ethmoidal nerve (AEN) and superior laryngeal nerve, respectively, in non-paralyzed decerebrate cats. Respiratory and laryngeal motor patterns that consisted of an inspiration phase, compression phase, and expulsion phase were observed for both sneezing and coughing. The main difference was observed in the activity of the elevator of the back of the tongue, styloglossus (SG) muscle, which was explosively activated during the expulsion phase of sneezing, whereas it was virtually silent during coughing. The nasopharyngeal closers were weakly to moderately activated during sneezing. Their activities during coughing were weaker than during sneezing. Furthermore, the AEN-induced activities of the phrenic and abdominal nerves and the lateral branch of the hypoglossal nerve (lat XII), which innervates the SG muscle, in paralyzed cats were consistent with the activities of the diaphragm, abdominal, and SG muscles during actual sneezing in non-paralyzed cats. Thus, we conclude that tongue movement is the main difference in the motor outputs between sneezing and coughing, which probably causes greater nasal airflow in sneezing, and that it is necessary to record the activity of the lat-XII to identify fictive sneezing in paralyzed cats. PMID- 9858021 TI - Neuronal degeneration and microglial reaction in the fetal and postnatal rat brain after transient maternal hypoxia. AB - This study examined the neuropathological changes in different areas of the brain of fetal and postnatal rats after transient maternal hypoxia. At different time intervals following hypoxia, reactive microglia as determined immunohistochemically with the antibody OX-42 that recognizes complement type three (CR3) receptors, responded vigorously to the hypoxic stress. Microglial activation was particularly evident in the cingulate cortex and the corpus callosum between 3 h and 14 days after hypoxia. Massive cell degeneration as determined ultrastructurally and significant neuronal loss as evaluated by cell counts were observed in the cingulate cortex at 1 and 3 days after hypoxic insults; thereafter, however, the neuronal density was restored to normal levels. Present results suggest that the cingulate cortex is most vulnerable to the hypoxic injury probably due to a redistribution of cerebral blood flow and/or metabolic changes. Besides being involved in the phagocytosis of cellular debris, it is suggested that the reactive microglial cells may have both neurotoxic and neurotrophic functions. PMID- 9858022 TI - Axonal regeneration with functional restoration in the vestibulospinal tract in young rats. AB - The present study examined regeneration and restoration of function of the mammalian central vestibular system in the infant rat. The lateral vestibulospinal tract (LVST) of rats was completely transected unilaterally by a ventral approach. After a postoperative interval of one day to three months, the LVST was examined by anterograde transport of wheat-germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) and retrograde transport of fluorescent dye. Twelve of the 22 LVST-transected rats showed successful regeneration. The regenerated fibers formed a compact fiber bundle, which sent terminals to their normal targets. The contribution of the regenerated fibers to functional recovery was estimated by analyzing the locomotor capacity of the transected rats. The locomotor movements were measured on the surface of a digitizer table by attaching a miniature resonance coil to the abdomen of the rats. Rats which shows normal locomotor movements represented a marked regeneration of LVST fibers. In contrast, rats with poorly-controlled locomotor movements showed unsuccessful regeneration. These results suggest that, contrary to previous thought, regeneration and functional restoration of the central vestibular system in young rats does occur. PMID- 9858023 TI - Adaptive modifications of post-saccadic smooth pursuit eye movements and their interaction with saccades and the vestibulo-ocular reflex in the primate. AB - Adaptation of the horizontal smooth pursuit eye movement was examined using step ramp moving target paradigms in chronically prepared Macaca fuscata. Monkeys were trained to pursue a small target which moved in the horizontal plane in a 3 degrees step-10 degrees/s ramp or a 0 degrees step-10 degrees/s ramp mode for 300 400 ms. When the target moved from central fixation point in a step-ramp mode, the monkeys usually responded with an initial pursuit eye movement (latency, 100 120 ms) which reached to a nearly constant velocity of 4 degrees/s in 50 100 ms, followed by a 1-3.5 degrees catch-up saccade (latency, 170-230 ms). The catch-up saccade was followed by a 7-8 degrees/s post-saccadic pursuit. The post-saccadic pursuit velocity was measured 40-90 ms after the end of the catch-up saccade. When the target velocity was doubled (20 degrees/s) for 100-200 ms immediately after the onset of the catch-up saccade, the post-saccadic pursuit velocity increased by 40%. When the target velocity was decreased by half (5 degrees/s) immediately after the onset of the catch-up saccade for 150 ms, the post-saccadic pursuit velocity decreased by 30%. These increases or decreases of post-saccadic pursuit were observable within just 50-150 trials. The adaptation of post saccadic pursuit occurred independent of the position of the target. The amplitude and latency of the catch-up saccade also increased correspondingly when the post-saccadic pursuit velocity was adaptively increased. Adaptation of smooth pursuit did not affect the dynamics of reflex eye movements, including the horizontal vestibulo-ocular reflex (HVOR) gain and phase measured by 0.33 Hz-10 degrees (peak-to-peak) turntable oscillations in darkness. Conversely, adaptation of the HVOR gain induced by a 2 h sustained oscillation of the turntable and screen in reversed direction at 0.33 Hz-10 degrees affected little the velocity of post-saccadic pursuit or the amplitude of the catch-up saccade. These results suggest that different neural mechanisms are respectively involved for the adaptation of horizontal smooth pursuit and HVOR in the primate. PMID- 9858024 TI - Inhibitory effects of ascorbic acid on dopamine uptake by rat striatal synaptosomes: relationship to lipid peroxidation and oxidation of protein sulfhydryl groups. AB - Ascorbic acid is frequently added in the incubation medium to prevent oxidation of dopamine (DA) during uptake assays. However, a preliminary study showed that the presence of ascorbic acid induced a decrease of DA uptake after prolonged incubation. The purpose of this study was to determine the mechanism underlying ascorbic acid-induced alterations of DA uptake in rat striatal synaptosomes. In this context, the effects of physiological concentrations of ascorbic acid (100 500 microM) on DA uptake and Na+/K+ ATPase activity (which is essential for DA transporter function) were assessed in synaptosomes before and after incubation at 37 degrees C. The capacity of synaptosomes to take up DA was significantly decreased after incubation owing to a reduction in DA transporters (but with no modification of their affinity for DA). This partial inhibition was associated with a decrease of Na+/K+ ATPase activity, a production of thiobarbituric acid reactive substances (TBARS) and malonaldehyde (MDA), and a loss of sulfhydryl group content. Addition of Trolox C to the medium prevented the reduction of DA uptake, the inhibition of Na+/K+ ATPase activity, the decrease in sulfhydryl group content and the production of TBARS and MDA. These results suggest that ascorbic acid in the presence of contaminant ferrous ions induced a decrease in functional DA transporters, probably through a lipid peroxidation process involving oxidation of sulfhydryl groups and at least in part through a decrease of Na+/K+ ATPase activity. PMID- 9858025 TI - Food-taking in the cat investigated with transection of the rubro- and corticospinal tracts. AB - In cats with the distal paw shaved food-taking from a horizontally or vertically placed tube was recorded with a video camera, supplementary results were obtained with a high frequency video system. Measurements were made of the dorsal contour of the digits to investigate angular movement in the proximal interphalangeal (PIP) joint and withdrawal of the paw before and after complete transection of the rubro- and corticospinal tracts just rostral to the forelimb segments. In the preoperative state PIP flexion occurs before withdrawal of the paw in the large majority of cats. Postsurgically the main part of PIP flexion is made during withdrawal of the paw. It is suggested that PIP flexion before withdrawal allows for accurate manipulative placement of the digits on the target which depends on the rubro- and corticospinal tracts, while PIP flexion during withdrawal, which may favour speed at the expense of accuracy, can be made without the rubro-and corticospinal tracts. PMID- 9858026 TI - Short-latency vergence eye movements elicited by looming step in monkeys. AB - The looming associated with forward and backward motion of the observer has been shown to elicit vergence eye movements with short-latency (approximately 80 ms) in human subjects. We studied the vergence eye movements elicited by looming in three monkeys (Macaca fuscata). The animals faced a large tangent screen onto which a random dot pattern was back-projected. The movements of both eyes were recorded with an electromagnetic induction technique. Fifty milliseconds after a centering saccade, this first pattern was replaced with a new one that showed the same image viewed from a slightly different distance. This looming step (two frame movie) included both radial optic flow and a size change. As expected from the optical geometry, centrifugal flow coupled with enlargement (signaling forward motion) increased the vergence angle, whereas the converse combination decreased the vergence angle. In both cases, the optimal step-change in apparent viewing distance was 2%. The latency of these vergence responses was very short and similar to those induced when disparity steps are applied to such large patterns (approximately 60 ms). We suggest that these two systems act in synergy to help maintain binocular alignment during forward and backward motion of the observer. PMID- 9858027 TI - CSF levels of HVA and 5-HIAA in drug-free schizophrenic patients and healthy controls: a prospective study focused on their predictive value for outcome in schizophrenia. AB - The CSF levels of HVA and 5-HIAA were determined in 90 drug-free DSM-III-R schizophrenic patients and 47 healthy control subjects, and their predictive value for 5-year outcome was evaluated. CSF was collected by lumbar puncture at index admission, and in 37 of the patients a second sample was drawn after approx. 7 weeks of neuroleptic treatment. Outcome was rated prospectively 5 years after index admission by means of the Strauss-Carpenter outcome scale. Schizophrenic patients had significantly lower levels of HVA in the CSF than the control group, but no difference was found for 5-HIAA. The CSF-amine metabolite levels were not correlated with age at admission, age at first symptoms or duration of the disorder. Neither HVA nor 5-HIAA correlated with the total outcome scores at a 1- and 5-year follow-up evaluation. First-admitted previously untreated patients with the poorest 5-year outcome had significantly lower HVA/5 HIAA quotients than those with a good outcome. Furthermore, patients still having a low HVA/5-HIAA quotient after treatment with neuroleptics had a poorer 5-year outcome than patients with an increased quotient. The data indicate that both HVA and 5-HIAA in the CSF, and especially their sensitivity to neuroleptic treatment, have a predictive value for the prognosis in schizophrenia. PMID- 9858028 TI - No linkage or linkage disequilibrium between brain-derived neurotrophic factor (BDNF) dinucleotide repeat polymorphism and schizophrenia in Irish families. AB - There is increasing evidence that a neurodevelopmental process is accountable for at least a proportion of schizophrenic cases. Brain-derived neurotrophic factor (BDNF), a member of a group of proteins that includes neurotrophin-3/4/5 and nerve growth factor (NGF), is an attractive candidate gene. We have performed a case control association study using the BDNF dinucleotide repeat polymorphism in a sample of familial schizophrenic individuals and in healthy, ethnically matched control subjects. We also performed a linkage analysis on 265 multiplex families using the same marker. We found no differences in allele frequencies between the patient and control groups nor any evidence for transmission disequilibrium or linkage with the multiply affected families. We conclude that DNA variation at or near the BDNF gene is unlikely to contribute to the genetic predisposition to schizophrenia. PMID- 9858029 TI - Functional polymorphism of -141C Ins/Del in the dopamine D2 receptor gene promoter and schizophrenia. AB - Several studies showed the density of D2 receptors was elevated in postmortem brains from schizophrenics. Genes which operate at the level of gene activation may be associated with the pathogenesis of schizophrenia. Arinami et al. [(1997) Human Molecular Genetics 6, 577-582] found a polymorphism in the 5'-flanking region of the D2 receptor gene designated as -141C Ins/Del. The promoter activity by luciferase assay of a plasmid containing the -141C Ins allele was higher than in the one containing the -141C Del allele. In addition, the -141C Ins allele frequency was significantly higher in schizophrenics than in control subjects. We replicated the -141C Ins/Del polymorphism in 170 schizophrenics and 121 healthy control subjects. The number of schizophrenics with the -141C Ins/Ins genotype was significantly higher than that of control subjects (P = 0.038). The frequency of the -141C Ins allele was significantly increased in the schizophrenics compared with the control subjects (P = 0.042). The mean age of onset for the patients with -141C Ins/Del was significantly lower than that for the patients with -141C Ins/Ins (P = 0.029). There was no association between the genotype and either positive symptoms or the response to antipsychotic medication. Our results suggest that the -141C Ins/Del polymorphism may affect the susceptibility to schizophrenia. PMID- 9858030 TI - Age at onset and gender resemblance in bipolar siblings. AB - In order to measure the intrafamilial correlation for age at onset and to examine gender resemblance among bipolar siblings, we assessed a sample of 130 bipolar patients belonging to 59 multiple affected sibships. To study the intrafamilial resemblance for age at onset and gender, we used the intraclass correlation and the sibship method, respectively. Within the whole sample, age at onset for affected siblings was correlated (rho = 0.42, P = 0.0001). Gender was randomly distributed among bipolar sibships, demonstrating the absence of gender resemblance among affected siblings. The existence of an intrafamilial correlation for age at onset among bipolar siblings suggests that this variable may assist in the identification of more heritable forms of the illness. No intrafamilial correlation was found for the gender of affected siblings, suggesting that familial vulnerability factors are not gender-specific. PMID- 9858031 TI - D2 dopamine receptor and GABA(A) receptor beta3 subunit genes and alcoholism. AB - As the dopaminergic and GABAergic systems have been implicated in alcohol-related behaviors, variants of the D2 dopamine receptor (DRD2) and GABA(A) receptor beta3 subunit (GABRB3) genes were determined in a population-based association study of Caucasian non-alcoholic and alcoholic subjects. In severe alcoholics, compared to non-alcoholics, a significant increase was found in the prevalence (P = 1.7 x 10( 5)) and frequency (P = 1.6 x 10(-5)) of the DRD2 minor (A1) allele. Moreover, a significant progressive increase was observed in A1 allelic prevalence (P = 3.1 x 10(-6)) and frequency (P = 2.7 x 10(-6)) in the order of non-alcoholics, less severe and severe alcoholics. In severe alcoholics, compared to non-alcoholics, a significant decrease was found in the prevalence (P = 4.5 x 10(-3)) and frequency (P = 2.7 x 10(-2)) of the GABRB3 major (G1) allele. Furthermore, a significant progressive decrease was noted in G1 allelic prevalence (P = 2.4 x 10(-3)) and frequency (P = 1.9 x 10(-2)) in non-alcoholics, less severe and severe alcoholics, respectively. In sum, in the same population of non-alcoholics and alcoholics studied, variants of both the DRD2 and GABRB3 genes independently contribute to the risk for alcoholism, with the DRD2 variants revealing a stronger effect than the GABRB3 variants. However, when the DRD2 and the GABRB3 variants are combined, the risk for alcoholism is more robust than when these variants are considered separately. PMID- 9858032 TI - Impulse-control disorders in alcoholics are related to sensation seeking and not to impulsivity. AB - Impulse-control disorders (ICD) include intermittent explosive disorder, kleptomania, trichotillomania, pyromania and pathological gambling. Several studies have suggested that the incidence of pathological gambling and impulsive violent behavior is substantially higher in alcohol-dependent patients than in the general population. The association between ICD and alcoholism, as well as personality characteristics such as sensation seeking and impulsivity, has never been systematically studied. The present study compared the levels of impulsivity and sensation seeking in age- and sex-matched groups of alcohol-dependent patients with concomitant ICD (ICD+, n = 30), alcohol-dependent patients without ICD (ICD-; n = 30) and control subjects (n = 30). All the alcohol-dependent patients (ICD+ and ICD-) were hospitalized for alcohol detoxification. Diagnoses of ICD were based on DSM-IV criteria and the Minnesota Impulsive Disorders Interview. All patients completed the Zuckerman Sensation-Seeking Scale (SSS) and the Barratt Impulsiveness Rating Scale (BIS). Mean scores on the SSS general factor, the SSS disinhibition subscale, and the SSS experience-seeking scale were significantly higher in ICD+ patients than in either ICD- patients or control subjects. By contrast, total scores and subscale scores on the BIS showed no significant differences among the three groups. Thus, it appears that measures of sensation seeking, rather than impulsivity, are relevant in distinguishing between alcohol-dependent patients with and without concomitant impulse control disorders. PMID- 9858033 TI - Phytohemagglutinin-stimulated calcium signal in lymphocytes of alcoholics before, during and after detoxification. AB - Acute and chronic exposure to ethanol influences intracellular calcium homeostasis via NMDA receptors, direct regulation of calcium channels or the phosphoinositide pathway. To explore the influence of alcohol withdrawal on calcium metabolism, we have investigated the resting and phytohemagglutinin (PHA) stimulated [Ca2+]i in lymphocytes of 10 alcoholics before, during and after withdrawal. Our findings suggest that both compartments of the PHA-stimulated signal are affected in alcoholics, with flattening of the initial peak and sustained calcium influx, as long as severe vegetative signs are present. MANOVA results showed significant interaction effects for both measurement points, for the initial peak, 40 s after stimulation (P = 0.05), and especially for the sustained influx at the end of the observation period (P = 0.001). The detailed mechanisms of this disturbed calcium homeostasis need further investigation. PMID- 9858034 TI - Dysregulation of cardiovascular and neuroendocrine responses to stress in premenstrual dysphoric disorder. AB - Twelve women with prospectively confirmed premenstrual dysphoric disorder (PMDD or PDD) were compared with 12 healthy control subjects for cardiovascular and neuroendocrine responses to speech and mental arithmetic (Paced Auditory Serial Addition Task) stressors during both the follicular and luteal phases of the menstrual cycle. Structured clinical interview was used to assess psychiatric and abuse histories, and standardized questionnaires were administered to assess current life stress. Results revealed that PMDD women had significantly lower stroke volume, cardiac output and cortisol levels but significantly elevated norepinephrine and total peripheral resistance at rest and also during mental stressors compared with control subjects. These effects were evident in both cycle phases. Significantly more women with PMDD had histories of sexual abuse, and they also reported greater current life stress than control subjects. Consistent with a history of trauma, the PMDD women exhibited significantly greater ratios of norepinephrine to cortisol at rest and during stress. These results are interpreted as reflecting dysregulation of the stress response and may be related to histories of severe and/or chronic exposure to stress for a subgroup of PMDD women. PMID- 9858035 TI - Epidemiologic and phenomenological aspects of post-traumatic stress disorder: DSM III-R diagnosis and diagnostic criteria not validated. AB - The aim of this cohort study was: (i) to validate the diagnostic criteria for post-traumatic stress disorder (PTSD) of the DSM-III-R; and (ii) to examine the incidence rate of PTSD in a study population exposed to two different traumatic events, i.e. a fire in a hotel ball-room and a multiple collision car-crash on a Belgian highway. One hundred and eighty-five victims (130 fire and 55 car accident victims) were assessed between 7 and 9 months after the traumatic event using the Composite International Diagnostic Interview (CIDI), PTSD Module, a fully structured diagnostic interview for the assessment of PTSD according to DSM III-R criteria. Twenty-three percent of the study population met DSM-III-R criteria for PTSD. By means of unsupervised and supervised multivariate statistical analyses we were unable to validate the three-factorial structure, i.e. criteria B, C and D, of the DSM-III-R PTSD diagnosis. The latter relies heavily on the C diagnostic criteria, which appear to be too restrictive. Women were more likely to develop symptoms of reexperience (B) and arousal (D) than men. There was a significantly higher incidence of criteria B, C and D, but not of PTSD, in fire than in car-accident victims. Between 42 and 57% of the victims developed the first PTSD symptoms on the day of the trauma; within the next week these incidence rates increased to 77.1, 57.8 and 73.5% for criteria B, C and D, respectively. In conclusion, this study was unable to demonstrate the validity of the diagnostic criteria for PTSD according to DSM-III-R. The present cohort study has defined a number of factors that may predict new occurrences of PTSD symptoms after a traumatic event, i.e. gender, type of trauma and time delay between the trauma and the assessment of the diagnostic criteria. PMID- 9858036 TI - The two-factorial symptom structure of post-traumatic stress disorder: depression avoidance and arousal-anxiety. AB - The first part of this study showed that the DSM-III-R symptom structure of post traumatic stress disorder (PTSD), i.e. criteria B (reexperience), C (avoidance numbing), and D (arousal), and, consequently the diagnosis of PTSD, could not be validated in fire and car-accident victims. The aims of this study were to: (i) determine the factors as well as their structure in the symptoms of PTSD; and (ii) develop a new classification or typology of PTSD. Exploratory and confirmatory factor analyses and cluster analyses were employed to: (i) examine the factors in PTSD symptomatology; and (ii) find and validate adequate diagnostic criteria for PTSD. The Composite International Diagnostic Interview (CIDI), PTSD Module, was used between 7 and 9 months after the traumatic event in a study group of 185 victims of two different traumatic events, i.e. 130 fire and 55 car-accident victims. Our findings support the existence of two factors, i.e. a first labeled 'depression-avoidance (DAV) dimension', as it contains items reminiscent of depression and avoidance, and a second labeled 'the anxiety arousal (AA) dimension', as it contains symptoms reminiscent of anxiety and increased arousal. Cluster analysis yielded two clusters, i.e. a cluster of subjects with PTSD cases and another with non-cases. Our PTSD algorithm was significantly less restrictive than the DSM-III-R diagnosis of PTSD. There are only quantitative, but no qualitative, differences between the cluster analytically derived classes. IN CONCLUSION: PTSD is not a well-delineated clinical entity, as there is a clinical continuum from PTSD non-cases to cases with less and more severe DAV and AA symptoms. It is more appropriate to express PTSD in terms of general severity of PTSD and severity of the DAV and AA dimensions. PMID- 9858037 TI - Familial transmission of seasonal changes in sleep and eating function in the general population. AB - Both genetic and environmental factors may be involved in the development of the increased sleep and appetite/body weight during winter, which are observed in seasonal affective disorder (SAD) as well as in normal variants of behavior in the general population. Decreases of these psychosomatic functions are also observed during summer, although the change may be less apparent than the winter changes. We studied familial correlation of these seasonal changes in 129 small Japanese families of the general population, using the Seasonality Pattern Assessment Questionnaire (SPAQ). No association was observed between the child and biological parents in the changes of sleep length or appetite/body weight during winter or summer. In contrast, significant spouse correlations were found in the changes of sleep and eating function during winter as well as during summer. This may indicate a role of shared environmental factors in the development of the seasonal patterns. The present results are preliminary and further studies are recommended. PMID- 9858038 TI - Seasonality in suicides: the influence of suicide method, gender and age on suicide distribution in Italy. AB - In order to ascertain if and how age, gender and choice of lethal means influence the seasonal distribution of suicide in Italy, data concerning all suicides registered in Italy from 1984 to 1995 have been analyzed, taking these variables into account. In the age group 14-65 years and over a total of 31771 male suicides (mean yearly rate, 12.6 per 100000) and 11984 female suicides (mean yearly rate, 4.4 per 100000) have been identified in Italy during the study period. Suicides in the younger age ranges, both among males and females, show a less marked asymmetrical seasonal distribution than those in the older age groups. Only suicides committed by violent methods (ICD 953-958) show clear evidence of seasonality, with a peak in spring and a low in late autumn. Suicides committed by non-violent methods (950-952) follow no seasonal trend in either sex. Spectral analysis reveals a circannual rhythm for violent suicides (ICD 953 958) in both genders. For male non-violent suicides (ICD 950-952), a period with a frequency of 0.0833 (12 months) has been identified, but with a polarity opposite to that of male violent suicides. For female non-violent suicides, no period of frequency of 0.0833 could be identified, but, as for female violent suicides, a period with frequency close to 0.2500 (4 months) has been found. Changes in climate, then, correlate with the monthly distribution of violent and non-violent suicides in opposite ways: male violent suicides show a significant positive relationship with indicators of temperature and exposure to the sun, and a significant negative relationship with indicators of humidity and rainfall. Female suicides show less significant relationships with climate indicators. Work aimed at suicide prevention should therefore take into account the complex influence of seasonal climate both on human biological rhythms (particularly on 5 HT related functions and their actions on mood and impulsivity) and on sociorelational habits. PMID- 9858039 TI - Decreasing tendency of seasonality in suicide may indicate lowering rate of depressive suicides in the population. AB - The seasonality of suicide is well known and a repeatedly demonstrated phenomenon. The authors analyzed the seasonality of 148 suicide events on Gotland between 1981 and 1996. A marked and significant seasonality with a spring and summer peak was found between 1981 and 1989, when the prescription of antidepressants was relatively low and stable. However, this seasonality disappeared in the period between 1990 and 1996, when prescription of antidepressants increased dramatically, indicating that more and more depressed patients were pharmacologically treated. As the seasonality of suicide in the population is the reflection of the seasonal nature of depressive suicides, the result suggests that a decreasing tendency of seasonality in suicide may indicate the lowering rate of depressive suicides in the given population. PMID- 9858040 TI - Positive and negative schizotypal symptoms relate to different aspects of crossover reaction time task performance. AB - Although the expressions of both positive and negative symptoms in schizophrenia spectrum illnesses can each occur with varying degrees of severity, researchers have often dichotomized patients as generally positive or negative subtypes. Studies of schizophrenia and schizotypal personality disorder (SPD) have not typically controlled for the severity of the other symptom types when examining the relationship between positive and negative symptom subtypes and cognitive impairment. The present study investigated the relationship between the severity of both symptom types and reaction time crossover task performance in SPD in groups made equivalent on the severity of the other type of symptom. Fifty-eight out of 458 undergraduates were screened into one of four groups (high negative high positive, low negative-low positive, high negative-low positive or low negative-high positive) by the Schizotypal Personality Questionnaire and assessed with the reaction time crossover task. The results indicated that negative schizotypal symptoms were associated with the early crossover pattern, while positive schizotypal symptoms related to longer overall reaction time. Therefore, different cognitive mechanisms involved in crossover task performance appeared to be associated with different symptom subtypes. PMID- 9858041 TI - Olfactory identification deficiency and WCST performance in men with schizophrenia. AB - Several studies using the University of Pennsylvania Smell Identification Test (UPSIT), which requires forced choice olfactory identifications, have reported olfactory identification deficits in patients with schizophrenia. This report examines the possible links between olfactory identification (usually attributed to the orbitofrontal cortex) and executive functions (usually attributed to dorsolateral prefrontal cortex) in 24 male patients with schizophrenia and 21 male comparison subjects. Olfactory performance was investigated under two conditions: spontaneous identification and forced choice identification. Executive function was assessed with the Wisconsin Card Sorting Test (WCST). Compared with controls, patients with schizophrenia exhibited a higher average number of cigarettes smoked per day, lower spontaneous identification scores on olfactory performance, and a higher percentage of perseverative errors on the WCST; there was a significant relationship between the two performance measures. Simpson-Angus scores, neuroleptic drug treatment levels, and scores on the Positive and Negative Syndrome Scale were not correlated with either olfactory measure. The lack of correlation between the forced choice olfactory identification score and the WCST score is consistent with findings in previous studies that used the UPSIT. By contrast, use of a spontaneous identification condition to assess olfactory performance did produce a significant association with WCST performance. PMID- 9858042 TI - Validity of the shortened Mood and Feelings Questionnaire in a community sample of children and adolescents: a preliminary research note. AB - The Mood and Feelings Questionnaire (MFQ) was designed to detect clinical depression in children and adolescents. Our aim was to investigate the relationship between symptom scores obtained using the short-version MFQ and psychiatric disorders in a non-clinical sample. Seventy-eight parents and 71 twins, who had completed the MFQ, were interviewed separately using a semistructured diagnostic interview, the Child and Adolescent Psychiatric Assessment. Parent-rated MFQ scores (MFQ-P) were found to distinguish those with ICD-10 (point biserial correlation = 0.345) and DSM-III-R depression (point biserial correlation = 0.369) from non-depressed cases. MFQ-P scores also differentiated depressed cases from those with 'other psychiatric diagnoses' (any anxiety disorder, oppositional defiant disorder and conduct disorder, hyperkinetic disorder/attention deficit hyperactivity disorder and adjustment disorder/post-traumatic disorder). The MFQ-P at the chosen cut-off point showed a sensitivity of 0.75 and specificity of 0.73 for an ICD-10 diagnosis of depression and a sensitivity of 0.86 and specificity of 0.87 for DSM-III-R depression. The number of self-rated reports (MFQ-C) was small, but overall the results suggest that self-rated MFQ scores may show less specificity. The MFQ-C at the selected cut-off point showed a sensitivity of 0.6 and specificity of 0.61 for ICD-10 depression, and a sensitivity of 0.75 and specificity of 0.74 for DSM-III-R depression. PMID- 9858043 TI - Test-retest reliability of the rat N40 auditory evoked response: preliminary data. AB - The N40 auditory evoked potential (EP) in rats has been used to develop an animal model for the study of sensory gating mechanisms. In an examination of the test retest reliability of sensory gating indices, derived from the rat N40 EP, six rats were tested four times daily for 5 consecutive days. A paired-click paradigm (S1-S2) was used, and both the S2/S1 amplitude ratio and the S1-S2 amplitude difference measures of sensory gating capacity were calculated. Based on the means of four daily trials, reliability coefficients were as follows; r = 0.69 for S1, and r = 0.66 for S2 amplitudes. A reliability coefficient of 0.59 was found for the S1-S2 difference measure, and one of 0.47 for the S2/S1 ratio measure. These results suggest that the N40 can be used to study sensory gating, but caution has to be exercised in interpreting the data particularly in a repeated measures design. PMID- 9858045 TI - Why is sleep deprivation an orphan drug? PMID- 9858044 TI - Association between anxiety disorders and a functional polymorphism in the serotonin transporter gene. AB - Recently, individuals with the short form of the serotonin transporter were found to be associated with neurotic characteristics. An association study on this polymorphism was performed in anxiety disorder patients and control subjects. The short form allele frequency in patients tended to be higher than that in control subjects (81.7 vs. 74.5%). Although it is difficult to ascribe significance to these 'tendencies', these data may suggest that variation of this functional polymorphism makes some contribution to anxiety disorders. PMID- 9858046 TI - Afferent receptors in the airways and cough. AB - The roles of airway rapidly adapting receptors (RARs) and of C-fibre receptors in the induction of cough are reviewed. It is concluded that, while there is substantial evidence that irritant receptors in the laryngeal wall and RARs in the tracheobronchial mucosa can cause cough, the evidence for such a similar direct role for C-fibre receptors is tenuous. Indeed there is accumulating evidence that the C-fibre receptors may cause apnoea and rapid shallow breathing, and also reflexly inhibit cough. However the C-fibre receptors may release tachykinins when stimulated, and these in turn may cause plasma extravasation from mucosal postcapillary venules. RARs are excited by increases in interstitial liquid volume, so C-fibre receptors may indirectly enhance cough via the RARs. PMID- 9858047 TI - Hans Loeschcke, Robert Mitchell and the medullary CO2 chemoreceptors: a brief historical review. AB - In the late 1950s, stimulated by reports from Leusen in Belgium and Winterstein in Germany on ventilatory responses to spinal fluid acid, Hans Loeschcke from Gottingen, and Robert Mitchell of the University of California in San Francisco were independently seeking the site of respiratory chemosensitivity to CO2 which they presumed to be mediated by cerebro-spinal fluid hydrogen ions. In 1960 Loeschcke came to San Francisco to join Mitchell for 3 months of intensive hunting for the site of action. This essay describes the events surrounding the localization of ventral medullary superficial (VMS) chemosensitivity to topical acidification, and some of their subsequent and largely independent work on the location, nature and function of this structure. The discovery led to a vast literature on all aspects of the regulation of respiration. PMID- 9858048 TI - Cytochromes and oxygen radicals as putative members of the oxygen sensing pathway. AB - This study applies biophysical methods like light absorption spectrophotometry of cytochromes, determination of NAD(P)H-dependent superoxide anion (O2-) formation and localisation of hydroxyl radicals (*OH) by 3-dimensional (3D) confocal laser scanning microscopy to reveal in human cells putative members of the oxygen sensing signal pathway leading to enhanced gene expression under hypoxia. A cell membrane localised non-mitochondrial cytochrome b558 seems to be involved as an oxygen sensor in the hepatoma cell line HepG2 in cooperation with the mitochondrial cytochrome b563 probably probing additionally metabolic changes. *OH the putative second messenger of the oxygen sensing pathway generated by a Fenton reaction could be visualized in the perinuclear space of the three human cell lines used. Substances like cobalt or the iron chelator desferrioxamine, which have been applied in HepG2 cells to mimic hypoxia induced gene expression, interact on various sides of the oxygen sensing pathway confirming the importance of b-type cytochromes and the Fenton reaction. PMID- 9858049 TI - Variability of vigilance and ventilation: studies on the control of respiration during sleep. AB - Ventilation is under metabolic as well as under behavioural control. This causes a complex interaction between states of 'vigilance' and respiration. This paper briefly summarizes sleep-related changes of respiration and presents an experimental study on the course of respiratory CO2-sensitivity during a whole night's sleep in ten healthy volunteers. The feedback control of breathing was challenged by continuous step changes of inspired CO2 every 7 min, resulting in 60, 3-step steady-state hypercapnic ventilatory responses (HCVR) per night in each subject. We analysed the variability of baseline ventilation and the effects of hypercapnia on ventilation with respect to sleep stages. There were only small differences in baseline PCO2 and ventilation between sleep stages, but a high variability of the slope of the CO2-response curves in the course of the night, ranging from 0.5 to 3.0 L min(-1) Torr(-1). The HCVR was significantly lower during REM sleep than during all stages of NREM sleep. Due to a compensatory left shift of the flattened CO2-response curves, however, ventilation at baseline CO2 as well as during slight hypercapnia varied much less than would be expected from the high variability of slopes. We conclude that the characteristics of the CO2 sensitive feedback control system of respiration, are highly variable during sleep, but due to offsetting effects, PCO2 and ventilation remain quite stable in the physiological range. PMID- 9858050 TI - A pontine-hypothalamic temperature difference correlated with cutaneous and respiratory heat loss. AB - The role of cutaneous and respiratory heat loss for selective brain cooling in different species is discussed and new experimental results from a comparative study are summarized. In three species (cat, rabbit and rat) the difference between pontine and hypothalamic temperatures was studied as a function of head heat exchanger vasomotion appraised by the difference between hypothalamic and ear pinna (cats and rabbits) or nasal mucosa (rats) temperatures during the behavioral states of wakefulness and slow wave sleep at an ambient temperature of 24+/-1 degrees C. The results show that: (i) the pontine-hypothalamic temperature difference is an useful indicator of selective brain cooling since it is positive and inversely correlated with the hypothalamic-ear pinna temperature difference in cats and rabbits and with the hypothalamic-nasal mucosa temperature difference in rats; (ii) respiratory heat loss prevails quantitatively over cutaneous heat loss in maintaining this difference. PMID- 9858051 TI - Endogenous carbon monoxide in control of respiration. AB - It is being increasingly appreciated that gas molecules such as nitric oxide (NO) function as chemical messengers in the nervous system. Recent studies suggest that carbon monoxide (CO) is another gas molecule that has similar biological actions as NO. The purpose of this article is to highlight the current information on the significance of endogenously generated CO in control of breathing. In mammalian cells, CO is generated during oxidative cleavage of heme by heme oxygenases (HO) and molecular oxygen is essential for this reaction. Two forms of HO have been identified including an inducible HO-1, that resembles stress-inducible protein HSP-32, and a constitutively expressed HO-2. HO-2 is expressed in many respiratory related neural structures including airway ganglion, carotid body, petrosal and nodose ganglia., nucleus of the tractus solitarius (nTS), and neurons of the rostral ventrolateral medulla (RVLM). Basal expression of HO-1 is either very low or even absent, but can be elevated during oxidative stress and hypoxia. Physiological studies have shown that CO might be of importance in vagally mediated contractions of airways. Several lines of evidence indicate that endogenously generated CO is a physiological modulator of the ventilatory response to hypoxia via its actions on carotid bodies and perhaps at brainstem neurons. In addition, CO might play a role in ventilatory adaptation to hypoxia, as low oxygen is a potent inducer of HO-1. Many of the neuronal structures that express HO also contain NOS, the enzyme that generates NO. Much remains to be studied on regulatory interactions between CO and NO and their impact on breathing. PMID- 9858052 TI - Respiratory sensations during heavy exercise in subjects without respiratory chemosensitivity. AB - Breathlessness arises from increased medullary respiratory center activity projecting to the forebrain (respiratory corollary discharge hypothesis). Subjects with congenital central hypoventilation syndrome (CCHS) lack the normal hyperpnea and breathlessness during hypercapnia. The corollary discharge hypothesis predicts that if CCHS subjects have normal hyperpnea during exercise, they will experience normal breathlessness during exercise. To test this, we studied four CCHS subjects and six matched controls during an exhausting constant load cycling test requiring substantial anaerobiosis. CCHS subjects rated significantly less breathlessness at the end of the test than controls, but ventilation (index of respiratory corollary discharge) was also somewhat lower in CCHS (not significant). In both groups, breathlessness increased disproportionately more than ventilation towards the end of exercise. These data failed to disprove the corollary discharge hypothesis of breathlessness, but do suggest that the relationship between ventilation and breathlessness is non linear and/or that projections of chemoreceptor afferents to the forebrain (presumed lacking in CCHS) is one source of breathlessness in normals. PMID- 9858053 TI - Methods and clinical significance of studying chemical drives. AB - Studying chemical control of ventilation implies evaluation of both chemoreceptor functions taken into account however the mechanical factors influencing the effector organs. The role of abnormal chemical drives has been demonstrated in COPD patients. More recently the role of abnormal chemical drives was studied during sleep. Absent or severely depressed drives may facilitate the development of central apneas and hypoventilation. High drives may lead to periodic breathing eventually with central apneas as well. Most intriguing therefore is the role of chemical drives in the pathogenesis of the obstructive and central sleep apnea syndrome. There is accumulating evidence that fluctuations in the drive to breathe may adversely affect the upper airway patency and facilitate upper airway closure and obstructive apneas. Interaction with chemical drives (eg by administration of acetazolamide) has been shown to improve central (and eventually also obstructive) sleep apnea. Studying chemical drives will probably be clinically useful in solving the complex mechanisms controlling ventilation during sleep in patients with and without underlying airway or lung disease. PMID- 9858054 TI - Mechanisms and clinicophysiological implications of the sniff- and gasp-like aspiration reflex. AB - Mechanical stimulation of the pharyngeal mucosa in cats and some other mammals evokes the 'aspiration reflex' (AR) characterized by rapid and strong inspiratory efforts not followed by active expirations. It resembles other spasmodic inspiratory acts such as the sniff, the gasp, and the sigh in several aspects, e.g. reflex or semireflex triggering from the upper airways, the sudden onset and termination of such inspirations, the massive recruitment, steep rise and high peak amplitude of inspiratory unit activity, analogous ventilatory pattern, and contribution to arousal. The similarity of these spasmodic acts is manifested mainly in enhanced speed and volume of inhalation, although of different intensity, which is determined by the varying degree of forced inspiratory activity and a concomitant inhibition of expiratory activity. The extent of the inspiratory dilation of the glottis and the timing and range of late-inspiratory and/or postinspiratory glottal narrowing modulate the depth of aspiration. Thus, the inhalation can be moderate as in sniffing, which provides a transfer of odorants to the olfactory mucosa. In AR the airstream is presumably strong enough to tear off the mechanical particles from the naso- and oropharynx and to convey them into the hypopharynx to allow their subsequent elimination by reflex swallowing or coughing. Prolonged glottal opening allows either the transfer of some additional air to the bronchi by sighing to prevent the development of atelectasis, or redistribution of a larger amount of fresh air into the lungs by gasping to support autoresuscitation. Should aspiration be a common effective component in these spasmodic processes, then the easily elicitable AR could be beneficial as a simple model for studying their properties in health and disease. PMID- 9858055 TI - Evidence for image-scanning eye movements during transitive inference. AB - Contrary to earlier work, recent studies have demonstrated a reduction in eye movements during the solution of tasks that seem to require visual imagery, relative to verbal tasks. The present study provides evidence that the nature of the visual imagery required by a task determines whether saccades are evoked and in which spatial pattern. In two experiments, subjects solved transitive inference problems with the relational terms left/right and above/below, while the horizontal and vertical EOG were recorded. Subjects made more horizontal and fewer vertical saccades while solving problems with the left/right terms than while solving identical problems with above/below. The results of silent counting tasks showed that the rate of subvocalization can also influence saccadic rate, especially in the horizontal plane, but cannot explain the eye-movement patterns observed during transitive inference. The results are discussed in terms of a motor theory of voluntary thinking. PMID- 9858056 TI - Sex differences in seasonal variations in P300. AB - Previous reports of seasonal variations in P300 were based on cross-sectional observations of subjects tested at different times of the year. In this study, we tested three groups of subjects in each of two seasons: winter and spring, spring and summer, and summer and winter. We found winter or spring maxima in auditory and visual P300 and visual slow wave. This pattern of results, with the amplitude of P300 being inversely related to the amount of sunlight in a season, supports the hypothesis that the allocation of processing resources varies across the seasons. Our results also suggest a trend for an increased sensitivity of women, as compared with men, to seasonal influences on P300. Although our findings do not provide strong evidence that P300 varies systematically as a function of season, seasonal factors appear to affect cognitive processing (as indexed by P300) differentially in men and women. PMID- 9858057 TI - Effects of inter- and intramodal selective attention to non-spatial visual stimuli: an event-related potential analysis. AB - Event-related potentials (ERPs) were recorded to trains of rapidly presented auditory and visual stimuli. ERPs in conditions in which subjects attended to different features of visual stimuli were compared with ERPs to the same type of stimuli when subjects attended to different features of auditory stimuli. This design permitted us to study effects of variations in both intramodal and intermodal visual attention on the timing and topography of ERP components in the same experiment. There were no indications that exogenous N110, P140 and N180 components to line gratings of high and low spatial frequencies were modulated by either intra- or intermodal forms of attention. Furthermore, intramodal and intermodal attention effects on ERPs showed similar topographical distributions. These combined findings suggest that the same neural generators in extrastriate occipital areas are involved in both forms of attention. Visual ERPs elicited in the condition in which subjects were engaged in auditory selective attention showed a large positive displacement at the occipital scalp sites relative to ERPs to attended and unattended stimuli in the visual condition. The early onset of this positivity might be associated with a highly confident and early rejection of the irrelevant visual stimuli, when these stimuli are presented among auditory stimuli. In addition, the later onset of selection potentials in the intramodal condition suggests that a more precise stimulus selection is needed when features of visual stimuli are rejected among other features of the same stimulus pattern, than when visual stimuli are rejected among stimuli of another modality. PMID- 9858058 TI - Repressive and defensive coping styles predict resting plasma endorphin levels in the elderly. AB - Jamner, L.D., Schwartz, G.E., 1986, Psychosom. Med. 48, 211-223, have proposed that repressive and defensive coping are associated with greater central endogenous opioid activity. They reasoned that high-defensive (HD) participants' attenuated distress, increased pain tolerances, attenuated somatic symptomatology, and accentuated reports of positive emotions are consistent with actions of centrally active opioid peptides. The present study assessed plasma beta-endorphin (END) levels, Marlowe Crowne Social Desirability Scale (MCSD), and Taylor Manifest Anxiety Scale (TMAS) scores in men (n = 6) and women (n = 20) between the ages of 59 and 79. Contrary to predictions, HD had lower plasma endorphin levels than did low-defensive (LD) participants. Raw MCSD scores correlated negatively with endorphin levels. Findings were significant for women only, which may have been due to the small sample of men. The results are discussed as they related to the hypothesis that defensiveness involves alteration of central opioid systems. PMID- 9858060 TI - Bradykinin stimulates prepubertal rat germ cell proliferation in vitro. AB - The effect of bradykinin on germ cell proliferation was studied by using in vitro organ cultures of testicular fragments from 3.5- and 4.5-day-old rats in the presence of 3H-thymidine. Bradykinin in tested concentrations of 10(-8), 10(-6) and 10(-4) M manifested a significant stimulation of rat prespermatogonial cell proliferation. The percentage of labeled germ cells increased up to 3-fold in comparison with the control value especially in 3.5-day-old rat testicular explants. Two kininase inhibitors: phosphoramidon and captopril were also used in combination with bradykinin. The percentage of labeled germ cells was slightly increased as compared to the samples treated with bradykinin only. Experiments with two agonists and two antagonists of B1 and B2 receptors for kinins showed that the rat prespermatogonial cell proliferation response to bradykinin is probably mediated via B2 receptors. The present findings suggest that bradykinin may be an important local testicular factor with paracrine or autocrine role in the regulation of spermatogonial cell proliferation and germ cell number. PMID- 9858059 TI - Anxiety and autonomic flexibility: a cardiovascular approach. AB - Autonomic characteristics of panickers, blood phobics, and nonanxious controls were compared with a variety of cardiovascular measures, including spectral analysis of the cardiac inter-beat interval time series (derived from the electrocardiogram). Responses to laboratory stressors (shock avoidance and cold face stress) of 16 participants who reported recent occurrences of frequent severe panic attacks, 15 participants who reported strong somatic reactions and fainting to the sight of blood, and 15 controls, were recorded. Results suggested distinct autonomic patterns among the three groups. Across conditions, panickers displayed the highest heart rates (HR) coupled with the least HR variability, which indicates low levels of cardiac vagal tone. Blood phobics showed more vagally mediated HR variability than panickers, with a significant association between cardiac rate and mean arterial pressure. Controls generally showed the most HR variability and 'spectral reserve' (a quality that indicates flexible responsivity). Results are discussed in the context of traditional models of anxiety and autonomic activity in contrast to contemporary notions of stability and change in biological systems. PMID- 9858061 TI - Delta9 tetrahydrocannabinol and cannabidiol alter cytokine production by human immune cells. AB - Marijuana, a widely abused drug in the US, and its derivatives (cannabinoids) have been used in AIDS and cancer patients for treatment of intractable nausea and cachexia. Yet, objective investigations of the effect of cannabinoids on the human immune system are few. We investigated the effect of delta9 tetrahydrocannabinol (THC) and cannabidiol (CBD) on cytokine production in vitro by human leukemic T, B, eosinophilic and CD8+ NK cell lines as models. THC decreased constitutive production of IL-8, MIP-1alpha, MIP-1beta, and RANTES and phorbol ester stimulated production of TNF-alpha, GM-CSF and IFN-gamma by NK cells. It inhibited MIP-1beta in HTLV-1 positive B-cells but tripled IL-8, MIP 1alpha and MIP-1beta in B-cells and MIP-1beta in eosinophilic cells but doubled IL-8. Both cannabinoids strongly inhibited IL-10 production by HUT-78 T-cells. Results indicate that THC and nonpsychotropic CBD have complex lineage and derivative specific effects on cytokines consistent with previous animal studies. These effects while of potential benefits in some inflammatory/autoimmune diseases may worsen HIV infection, tumorigenesis and allergic inflammation in the lung. PMID- 9858062 TI - Induction of immunomodulating cytokines by a new polysaccharide-peptide complex from culture mycelia of Lentinus edodes. AB - The immunomodulating effect of a new polysaccharide-peptide complex from culture mycelia of Lentinus edodes (LE) was studied for elucidation of the mechanism of augmentation of cell-mediated immunity. RNA samples were isolated from the untreated and treated murine splenocytes and human peripheral blood mononuclear cells. RT-PCR was used to analyze the cytokine gene expression and bioassay was used to analyze the cytokine production. By administration of LE, the expression levels of IL-2 and TNF-alpha genes were augmented in the treated murine spleen mononuclear cells and human peripheral blood mononuclear cells. The production of IL-2 were augmented in the treated murine spleen mononuclear cells, and the production of TNF-alpha were augmented in the treated murine peritoneal exudate macrophages. The production of IL-2 and TNF-alpha were augmented in the treated human peripheral blood mononuclear cells. These results suggest that LE may induce Th immune responses. PMID- 9858063 TI - The influence of base sequence on the immunological properties of defined oligonucleotides. AB - To assess the influence of base sequence on the immunostimulatory activities of DNA, cell binding and mitogenicity of a series of 30-mer phosphodiester oligonucleotides were tested using murine spleen cells. These compounds consisted of either a single base or a six base CpG motif in the context of 5' and 3' flanking sequences of each base. Among fluoresceinated oligonucleotides, (dG)30 had the highest binding of single base compounds tested while the presence of dG flanks increased binding of compounds with six base motifs, whether active on inactive. In assays of mitogenesis including incorporation of thymidine and uridine as well as expression of cell surface CD69, (dG)30 induced the highest responses among single base compounds. Among compounds with an active six base motif, the extent of proliferation varied with flanking sequence, with dG flanks producing the greatest stimulation in all assays tested. Together, these findings indicate that a variety of base sequences may affect the immunomodulatory properties of DNA, with the activity of dG sequences perhaps resulting from the formation of variant DNA structures. PMID- 9858064 TI - The effects of nonsteroidal anti-inflammatory drugs on immune functions of human peripheral blood mononuclear cells. AB - It is generally accepted that the nonsteroidal anti-inflammatory drugs (NSAIDs) exhibit anti-inflammatory effects primarily through inhibition of prostaglandin (PG) synthesis. However, effects of NSAIDs on immune responses are not fully understood. This study investigated effects of indomethacin and a new NSAID (d-2 [4-(3-methyl-2-thienyl)phenyl]propionic acid, termed as M-5011 in this study) on cytokine production, lymphocyte proliferation, activities of natural killer (NK) and lymphokine activated killer (LAK) cells and secretion of immunoglobulin (Ig). Both indomethacin and M-5011 augmented interleukin (IL)-2 production, whereas they suppressed IL-6 production both at the protein and mRNA levels. These two NSAIDs augmented proliferation of phytohemagglutinin (PHA)-stimulated PBMC and enhanced NK and LAK cell activities. In contrast, indomethacin was more potent than M-5011 in inhibition of both PG synthesis and Ig secretions by pokeweed mitogen (PWM)-stimulated PBMC. These results suggest that these two NSAIDs equally augment cell-mediated immunity, whereas indomethacin was more potent than M-5011 in the inhibition of humoral immunity and PG synthesis. PMID- 9858065 TI - Direct action of a protein-bound polysaccharide, PSK, on transforming growth factor-beta. AB - We investigated the action of a protein-bound polysaccharide, PSK, on transforming growth factor-beta (TGF-beta). (1) In in vitro-mixed culture of peripheral blood mononuclear cells (PBMC) from healthy human and mitomycin C treated human colon cancer cells, PSK or polyclonal antibody to TGF-beta significantly enhanced incorporation of 3H-thymidine into PBMC, and apparently decreased TGF-beta1 levels of acid-treated culture supernatant. (2) PSK or the antibody interfered with the quantitation by enzyme immunoassay of TGF-beta1 in acid-treated supernatant of the mixed culture. (3) PSK was suggested to form a complex with 125I-human recombinant TGF-beta1 standard, when changes in molecular weight of radioactivities were assessed by gel filtration. Recombinant human TGF beta1 inhibited growth of mink lung epithelial cell line Mv1Lu and promoted collagen synthesis in rat kidney fibroblast cell line NRK49F, but the complex did not have such activities. (4) In addition to TGF-beta1, PSK bound with TGF-beta2 and platelet-derived growth factor; however, PSK did not bind with 22 other species of cytokines and growth factors. (5) Protein moiety of PSK is suggested to play an important role in the expression of the activity. These results suggest that PSK modulates the biological activity of TGF-beta1 by binding to its active form. PMID- 9858066 TI - Antigen specific and nonspecific modulation of the immune response by aminolevulinic acid based photodynamic therapy. AB - Photosensitizers used normally in treating cancers have considerable potential for treatment of other diseases. One such photosensitizer is the endogenously synthesized photosensitizer protoporphyrin IX (PpIX). To better understand how protoporphyrin might be used in transplantation or in treating autoimmune diseases, information must be obtained on how the photosensitizer affects all immune cells. We used a combination of flow cytometry and in vitro activation assays (recall assays and mixed-lymphocyte reactions) to examine the effects of PpIX on the antigen specific component, lymphocytes and the non-antigen specific component, the macrophages/monocytes and dendritic cells of the immune system. Whereas, lymphocytes accumulate PpIX only when activated, both macrophages and dendritic cells accumulated PpIX immediately, without in vitro activation, as measured by flow cytometry. ALA-PDT (aminolevulenic acid-photodynamic therapy) treated adherent cells in the recall assay had a decreased capability to activate lymphocytes. By increasing the light dose in the recall assay, antigen primed lymphocytes were selectively eliminated from a population of cells. Stimulator cells in an MLR had a decreased stimulatory capacity following ALA-PDT treatment. Functional alterations are seen in both the antigen specific PMID- 9858067 TI - Alcoholism and drug abuse in three groups--bipolar I, unipolars and their acquaintances. AB - OBJECTIVE: Previous work has shown that manic-depressive illness and alcohol abuse are linked. This study further explores the relationship of alcohol and drug abuse in bipolar I patients and unipolar depressives and a comparison group obtained through the acquaintance method. METHOD: Diagnosis was accomplished according to Research Diagnostic Criteria (RDC): controls = 469; bipolars = 277; unipolar depressives = 678. Systematic data were gathered using the SADS on lifetime and current drug abuse and alcoholism. Both patients and comparison subjects were then followed prospectively for 10 years. First degree family members were interviewed using the RDC family history method. RESULTS: The group of bipolar patients and the group of unipolar patients had higher rates of drug and alcohol abuse than the comparison group when primary and secondary affective disorder patients were combined. However, primary unipolar patients did not have higher rates of alcohol or drug abuse than the comparison group. In contrast, primary bipolar patients had higher rates of alcoholism, stimulant abuse, and ever having abused a drug than the primary unipolar group and the control group. In an evaluation of the bipolar patients, drug abusers were significantly younger at intake and had a significantly younger age of onset of bipolar disorder. There was a significant increase in family history of mania or schizoaffective mania in the drug-abusing bipolar patients as compared to the non-abusing bipolar patients. LIMITATION: As in all adult samples of patients with affective illness, the chronology of alcohol and substance problems vis-a-vis the onset of illness was determined retrospectively. CONCLUSIONS: (1) Alcoholism and drug abuse are more frequent in bipolar than unipolar patients. (2) The drug abuse of bipolar patients tends toward the abuse of stimulant drugs. (3) In a bipolar patient, familial diathesis for mania is significantly associated with the abuse of alcohol and drugs. (4) More provocatively, these findings suggest the hypothesis of a common familial-genetic diathesis for a subtype of bipolar I, alcohol and stimulant abuse. CLINICAL IMPLICATIONS: The present analyses, coupled with two previous ones from the CDS, suggest that drug abuse may precipitate an earlier onset of bipolar I disorder in those who already have a familial predisposition for mania. Furthermore, in dually diagnosed patients with manic-depressive and alcohol/stimulant abuse history, mood stabilization of the bipolar disorder represents a rational approach to control concurrent alcohol and drug problems, and should be studied in systematic controlled trials. PMID- 9858068 TI - Evidence supporting the independent inheritance of primary affective disorders and primary alcoholism in the families of bipolar patients. AB - BACKGROUND: This study explored the nature of the association between bipolar disorder and alcoholism. METHODS: The authors studied 814 first-degree relatives of 121 bipolar patients, divided on the basis of response to lithium prophylaxis. Logistic regression analysis was used to analyze the contribution of demographic, familial and clinical variables to the risk of primary alcoholism in the relatives. RESULTS: The risk of primary alcoholism in relatives was not related to the degree of affective loading in the family or to the proband's lithium response. CONCLUSION: This study does not support a shared genetic liability between bipolar disorder and alcoholism. LIMITATIONS: This study lacked a control group, but the analysis accounted for this. CLINICAL RELEVANCE: These disorders are not alternative forms of the same illness. PMID- 9858070 TI - Bipolar I affective disorder: predictors of outcome after 15 years. AB - BACKGROUND: Robust predictors of long-term outcome in bipolar affective disorder would have substantial importance to both clinicians and researchers. Such predictors are not available, however, perhaps because of the limitations of previous efforts to find them. METHODS: In this study, 113 patients with bipolar affective disorder were followed semiannually for 5 years and annually for a subsequent 15 years. Of these, 23 (20.4%) had a poor long-term outcome indicated by the presence of mania or major depressive disorder throughout the 15th year. RESULTS: Among the baseline demographic and clinical variables tested, only active alcoholism and low levels of optimum functioning in the preceding 5 years characterized poor outcome patients. The persistence of depressive symptoms in the first 2 years of follow-up predicted depressive symptoms 15 years later but the early persistence of manic symptoms seemed to have no predictive value. A regression analysis eliminated alcoholism as an independent predictor. Thus, only poor optimal functioning in the 5 years before baseline assessment, and the persistence of depressive symptoms in the two subsequent years, were independently associated with poor, long-term prognosis. LIMITATIONS: Patients were recruited at tertiary care centers and sampling was therefore biased toward greater severity and chronicity. As is true of all naturalistic studies of course, treatment was not controlled. CONCLUSION: These findings suggest the existence of a poor outcome, depression-prone subtype of bipolar affective disorder. PMID- 9858069 TI - Major depressive disorder: a prospective study of residual subthreshold depressive symptoms as predictor of rapid relapse. AB - BACKGROUND: The study tested whether level of recovery from major depressive episodes (MDEs) predicts duration of recovery in unipolar major depressive disorder (MDD) patients. METHODS: MDD patients seeking treatment at five academic centers were followed naturalistically for 10 years or longer. Patients were divided on the basis of intake MDE recovery into residual depressive symptoms (SSD; N=82) and asymptomatic (N=155) recovery groups. They were compared on time to first episode relapse/recurrence, antidepressant medication, and comorbid mental disorders. Recovery level was also compared to prior history of recurrent MDEs ( > 4 lifetime episodes) as a predictor of relapse/recurrence. RESULTS: Residual SSD compared to asymptomatic recovery patients relapsed to their next MDE > 3 times faster (median=68 vs. 23 weeks) and to any depressive episode > 5 times faster (median=33 vs. 184 weeks). Residual SSD recovery status was significantly associated with early episode relapse (OR=3.65) and was stronger than history of recurrent MDEs (OR=1.64). Rapid relapse in the SSD group could not be attributed to higher comorbidity or lower antidepressant treatment. LIMITATIONS: Although inter-rater agreement on weekly depressive symptom ratings was very high (ICC > 0.88), some error may exist in assigning recovery levels. Antidepressant treatments were recorded, but were not controlled. CONCLUSIONS: MDE recovery is a powerful predictor of time to episode relapse/recurrence. Residual SSD recovery is associated with very rapid episode relapse which supports the idea that SSD is an active state of illness. Asymptomatic recovery is associated with prolonged delay in episode recurrence. These findings of this present study have important implications for the goals of treatment of MDD and for defining true MDE recovery. PMID- 9858072 TI - Cycloid psychoses predominate in severe postpartum psychiatric disorders. AB - BACKGROUND: The nosological status of postpartum psychoses has remained controversial because of their often 'atypical' symptomatology. A polydiagnostic approach may further clarify this issue. METHODS: In a retrospective study, we applied the ICD-10 and Leonhard's classification to 39 patients with severe postpartum psychiatric disorders. The patients were personally reexamined on average 12.5 years (6-26 years) after the onset of the illness. RESULTS: An acute onset and a polymorphous psychotic symptomatology with rapid changes characterized the majority of our cases. Unipolar depressive disorders (28%) and acute polymorphous psychotic disorders (21%) represented the largest proportions within the ICD-10-classification. Applying Leonhard's classification, over half the patients (54%) suffered from a cycloid psychosis. Among cycloid psychoses, motility psychoses clearly predominated. Schizophrenias occurred rarely (10%) according to both classifications. LIMITATIONS: Due to the unknown prevalence of the various diagnoses among women of child-bearing age, it is impossible to statistically infer a specific association between childbirth and a distinct diagnosis from our data. CONCLUSIONS: Our findings suggest that cycloid psychoses, in particular motility psychoses, account for the majority of postpartum psychoses, and do not support the hypothesis of a nosological independence of postpartum psychoses. PMID- 9858071 TI - Pre- and perinatal complications and risk for bipolar disorder: a retrospective study. AB - BACKGROUND: Many studies have reported that obstetric complications are risk factors for schizophrenia, but few studies have examined whether complications increase risk for bipolar disorder. METHODS: Bipolar-disorder probands and their adult siblings were diagnosed using DSM-III-R criteria. Obstetrical data from maternal reports were scored, blind to diagnosis, applying published scales that take into account number and severity of complications. RESULTS: Obstetric complication scores were significantly worse in probands than siblings without mood disorders. LIMITATIONS: Probands had relatively severe symptoms; research using more heterogeneous samples is needed. CONCLUSION: Results suggest obstetric complications are etiologically significant in bipolar disorder. PMID- 9858073 TI - Failure to demonstrate parent-of-origin effect in transmission of bipolar II disorder. AB - BACKGROUND: Parent-of-origin effect (POE) is suggested in transmission of bipolar disorder. Bipolar II disorder (BPII) should be considered separately. METHODS: The gender difference of transmitting parents, prevalence rate in children, and age at onset of patients in relation to the sex of the transmitting parent, were examined in 220 BPII patients. RESULTS: No evidence suggesting involvement of POE was found. CONCLUSION: POE is not involved in transmission of BPII. LIMITATION: Number of subjects is not sufficient. Rate of interviewed subjects differs between mothers and fathers. CLINICAL RELEVANCE: Female BPII patients do not transmit the disease more often than male patients. PMID- 9858074 TI - The emerging epidemiology of hypomania and bipolar II disorder. AB - The literature on the lifetime prevalence of the bipolar spectrum suggests rates of 3-6.5%. The Zurich cohort study identified a prevalence rate up to age 35 of 5.5% of DSM-IV hypomania/mania and a further 2.8% for brief hypomania (recurrent and lasting 1-3 days). The validity of DSM-IV hypomania and brief hypomania was demonstrated by a family history of mood disorders, a history of suicide attempts and treatment for depression. Comorbidity with anxiety disorders and substance abuse was found equally in both subtypes of hypomania. The study suggests that recurrent brief hypomania belongs to the bipolar spectrum. The findings should be verified on larger national cohorts in other epidemiological and clinical studies. PMID- 9858075 TI - The prevalence of major depressive and bipolar disorders in Hungary. Results from a national epidemiologic survey. AB - In order to estimate the prevalence of affective disorders in Hungary a sample of the Hungarian adult population (18-64 years) selected at random was interviewed using the Diagnostic Interview Schedule (DIS) which generated DSM-III-R diagnoses. The lifetime rate for Major Depressive Disorder (MDD) was 15.1%, and for Bipolar Disorders (BD) 5.1%. The female-to-male ratio was 2.7 for MDD and nearly equal for BD. The 1-year and 1-month period prevalence rates were 7.1% and 2.6% for MDD and 0.9% and 0.5% for manic episodes. A higher rate of divorced or separated persons was found among individuals with a lifetime diagnosis of MDD. Besides these, the lifetime diagnosis of BD coexisted with higher rates of the never-married state. The highest hazard rate for the development of BD or MDD was in the range 15-19 years but in MDD another peak was also found in the range 45 50 years. The first peak was characteristic of the recurrent, and the other one of the single form of MDD. Insomnia, loss of energy, decreased interest, concentration problems were the most common symptoms during the depressive episode, independent of polarity. Higher rates of lifetime diagnosis of dysthymia and all kinds of anxiety disorder were revealed among persons with MDD. BD was associated with GAD (Generalized Anxiety Disorder), and panic disorder more often than chance. PMID- 9858076 TI - Systematic clinical methodology for validating bipolar-II disorder: data in mid stream from a French national multi-site study (EPIDEP). AB - BACKGROUND: This paper presents the methodology and clinical data in mid-stream from a French multi-center study (EPIDEP) in progress on a national sample of patients with DSM-IV major depressive episode (MDE). The aim of EPIDEP is to show the feasibility of validating the spectrum of soft bipolar disorders by practising clinicians. In this report, we focus on bipolar II (BP-II). METHOD: EPIDEP involves training 48 French psychiatrists in 15 sites; construction of a common protocol based on the criteria of DSM-IV and Akiskal (Soft Bipolarity), as well as criteria modified from the work of Angst (Hypomania Checklist), the Ahearn-Carroll Bipolarity Scale, HAM-D and Rosenthal Atypical Depression Scale; Semi-Structured Interview for Evaluation of Affective Temperaments (based on Akiskal-Mallya), self-rated Cyclothymia Scale (Akiskal), family history (Research Diagnostic Criteria); and prospective follow-up. RESULTS: Results are presented on 250 (of the 537) MDE patients studied thus far during the acute phase. The rate of BP-II disorder which was 22% at initial evaluation, nearly doubled (40%) by systematic evaluation. As expected from the selection of MDE by uniform criteria, inter-group comparison between BP-II vs unipolar showed no differences on the majority of socio-demographic parameters, clinical presentation and global intensity of depression. Despite such uniformity, key characteristics significantly differentiated BP-II from unipolar: younger age at onset of first depression, higher frequency of suicidal thoughts and hypersomnia during index episode, higher scores on Hypomania Checklist and cyclothymic and irritable temperaments, and higher switching rate under current treatment. Eighty-eight percent of cases assigned to cyclothymic temperament by clinicians (with a cut off of 10/21 items on self-rated cyclothymia) were recognized as BP-II. Evaluation of this temperament by clinician and patient correlated at a highly significant level (r=0.73; p <0.0001). Cyclothymia and hypomania were also correlated significantly (r=0.51; p < 0.001). LIMITATION: In a study conducted in diverse clinical settings, it was not possible to assure that clinicians making affective diagnoses were blind to the various temperamental measures. However, bias was minimized by the systematic and/or semi-structured nature of all evaluations. CONCLUSION: With a systematic search for hypomania, 40% of major depressive episodes were classified as BP-II, of which only half were known to the clinicians at study entry. Cyclothymic temperamental dysregulation emerged as a robust clinical marker of BP-II disorder. These data indicate that clinicians in diverse practice settings can be trained to recognize soft bipolarity, leading to changes in diagnostic practice at a national level. PMID- 9858077 TI - Gender, temperament, and the clinical picture in dysphoric mixed mania: findings from a French national study (EPIMAN). AB - BACKGROUND: This research derives from the French national multisite collaborative study on the clinical epidemiology of mania (EPIMAN). Our aim is to establish the validity of dysphoric mania along a "spectrum of mixity" extending into mixed mania with subthreshold depressive manifestations; to demonstrate the feasibility of obtaining clinically meaningful data on this entity on a national level; and to characterize the contribution of temperamental attributes and gender in its origin. METHODS: EPIMAN involves training 23 French psychiatrists in four different sites, representing four regions of France; to rigorously apply a common protocol deriving from the criteria of DSM-IV and McElroy et al.; the use of such instruments as the Beigel-Murphy, Ahearn-Carroll, modified HAM-D; and measures of affective temperaments based on the Akiskal-Mallya criteria; obtaining data on comorbidity, and family history (according to Winokur's approach as incorporated into the FH-RDC); and prospective follow-up for at least 12 months. The present report concerns the clinical and temperamental features of 104 manic patients during the acute hospital phase. RESULTS: Dysphoric mania (DM defined conservatively with fullblown depressive admixtures of five or more symptoms) occurred in 6.7%; the rate of dysphoric mania defined broadly (DM, presence of > or = 2 depressive symptoms) was 37%. Depressed mood and suicidal thoughts had the best positive predictive values for mixed mania. In comparison to pure mania (0-1 depressive symptoms), DM was characterized by female over representation; lower frequency of such typical manic symptomatology as elation, grandiosity, and excessive involvement; higher prevalence of associated psychotic features; higher rate of mixed states in first episodes; and complex temperamental dysregulation along primarily depressive, but also cyclothymic, and irritable dimensions; such irritability was particularly apparent in mixed mania at the lowest threshold of depressive admixtures of two symptoms only. LIMITATION: In a study involving hospitalized affectively unstable psychotic patients, it was difficult to assure that psychiatrists making the clinical diagnoses would be blind to the temperamental measures. However, bias was minimized by the systematic and/or semi-structured nature of all evaluations. CONCLUSIONS: Mixed mania, defined cross-sectionally by the simultaneous presence of at least two depressive symptoms, represents a prevalent and clinically distinct form of mania. Subthreshold depressive admixtures with mania actually appear to represent the more common expression of dysphoric mania. Moreover, an irritable dimension appears to be relevant to the definition of the expression of mixed mania with the lowest threshold of depressive symptoms. Neither an extreme, nor an endstage of mania, "mixity" is best conceptualized as intrusion of mania into its "opposite" temperament - especially that defined by lifelong depressive traits - and favored by female gender. These data suggest that reversal from a temperament to an episode of "opposite" polarity represents a fundamental aspect of the dysregulation that characterizes bipolar disorder. In both men and women with hyperthymic temperament, there appears "protection" against depressive symptom formation during a manic episode which, accordingly, remains relatively "pure". Because men have higher rates of this temperament, pure mania is overrepresented in men; on the other hand, the depressive temperament in manic women seems to be a clinical marker for the well-known female tendency for depression, hence the higher prevalence of mixed mania in women. PMID- 9858078 TI - Signs and symptoms of mania in pure and mixed episodes. AB - BACKGROUND: Debate continues about the diagnosis of mixed mania and the restrictiveness of the DSM-III-R and DSM-IV criteria for Bipolar Disorder, mixed. Although awareness of dysphoric features during mania continues to grow, standard mania rating instruments do not adequately assess mixed states and there is a striking disparity between the dysphoric signs and symptoms emphasized in research studies and the commonly employed DSM criteria. METHODS: Three hundred sixteen inpatients meeting DSM-III-R criteria for Bipolar Disorder, manic or mixed, were evaluated by rating 20 signs and symptoms. The frequencies of these signs and symptoms were computed for both diagnostic subtypes and compared using chi2 statistics and conditional probability parameters. RESULTS: The most frequently noted signs and symptoms in mania are motor activation, accelerated thought process, pressured speech and decreased sleep. Although euphoric mood was present in a large portion of the cohort, irritability, dysphoric mood and mood lability were also prominent in the entire cohort. Dysphoric mood, mood lability, anxiety, guilt, suicidality, and irritability were the only symptoms significantly more common in the mixed group. In contrast, grandiosity, euphoric mood, and pressured speech were significantly more often observed in the pure manic group. Contrary to popular belief, paranoia did not differ significantly between the two groups. Suicidality was present in a non-trivial 7% of the entire cohort, including some subjects who did not meet the criteria for mixed mania. LIMITATIONS: The comparison of mixed and manic episodes requires the appropriate definition of mixed states. In the current report we use the DSM-III-R definition of Bipolar Disorder, mixed, which may be too rigid. CONCLUSIONS: The data underscore that mania is not a purely euphoric state. Substantial rates of dysphoria, lability, anxiety and irritability were noted in the "pure" manic patients, as well as in those who meet the full DSM criteria for Bipolar Disorder, mixed, suggesting, that perhaps a less restrictive definition of mixed states would be more appropriate. PMID- 9858079 TI - Social anxiety, hypomania and the bipolar spectrum: data, theory and clinical issues. AB - Reports in the literature indicate a subtle but consistent relationship between panic and bipolar II disorder. The possible connection between social phobia and bipolarity is less investigated. When we studied the treatment outcome of 32 social phobic patients administered either the reversible monoamine oxidase inhibitor (RIMA) meclobomide or the irreversible inhibitor MAOI phenelzine, we found that eighteen had remission > 50% of their socially anxious symptoms. Moreover, 14/18 of those improved became hypomanic, according to the Raskin Mania Scale (RMS) and the Young Mania Scale (YMS) coupled with expert clinical diagnosis. These findings possibly allude to a relationship of social phobia to bipolarity. Treatment with RIMA or MAOI exposed these subjects as having an atypical bipolar syndrome which is part of the bipolar spectrum. We then compared this special subset of subjects to the 18 socially phobic patients who failed to respond to RIMA's or MAOI's and to 26 patients with generalized anxiety disorder (GAD). Eleven of the 14 hypomanic responders gave histories of serious developmental deprivation (anaclisis); only 5/18 social phobics and 3/26 GADs without hypomanic responses had anaclitic histories. The author raises the possibility that anaclisis may have interacted with the impediment of volition of uncomplicated bipolar depression to produce social inhibition and anxiety. Finally, the author upholds the central role of depressive inhibition in bipolar disorder, which during antidepressant therapy often overshoots in a hypomanic direction; even in the absence of prior spontaneous hypomania, such disinhibition should classify this special subset of social phobic patients within the bipolar spectrum. PMID- 9858080 TI - The high prevalence of bipolar II and associated cyclothymic and hyperthymic temperaments in HIV-patients. AB - BACKGROUND: Although recent studies have shown high rates of current and lifetime depression in HIV-infected patients, there is little systematic data on the occurrence of bipolarity in these patients. METHOD: We compared 46 HIV patients with index major depressive episode (MDE) to an equal number of age- and sex matched seronegative MDE patients, and systematically examined rates of DSM-III-R bipolar subtypes (enriched in accordance with Akiskal's system of classifying soft bipolar disorders). RESULTS: Although HIV and psychiatric clinic patients had comparable background in terms of familial affective loading, HIV patients had significantly higher familial rates for alcohol and substance use. The more important finding was the significantly higher proportion of HIV patients with lifetime bipolar II disorder (78%), and associated cyclothymic (52%) and hyperthymic (35%) temperaments; the findings were the same irrespective of HIV risk status (intravenous drug user vs. homosexual and other risk groups combined). LIMITATIONS: The major methodologic limitation of our study is that clinicians evaluating temperament were not blind to affective diagnoses and family history. The comparison affective group was a sample of convenience drawn from the same tertiary care university facility. CONCLUSION: The finding of a high rate of bipolar II disorder in HIV patients has treatment implications for seropositive patients presenting with depression. More provocatively, we submit that premorbid impulsive risk-taking traits associated with cyclothymic and hyperthymic temperaments may have played an important role in needle-sharing drug use and/or unprotected sexual behavior, leading ultimately to infection with HIV. Given their public health importance, these clinical findings and insights merit further investigation. In particular, systematic case-control studies, as well as other large scale studies with prospective methodology need to be conducted. PMID- 9858081 TI - Antidepressants and suicide mortality. AB - BACKGROUND: This paper presents a nationwide analysis of suicide mortality in Finland from 1990 to 1995, when the total use of antidepressants, especially that by selective serotonin reuptake inhibitors (SSRIs) expanded in the country. METHODS: Suicide rate was analysed by various methods including that by intake of antidepressants. Various antidepressants were compared by calculating fatal toxicity indices (FTI) by relating number of fatal poisonings by a drug to its consumption. RESULTS: The expanded use of antidepressants coincided with an increased number of deaths caused by these drugs. The proportion of suicides committed by use of antidepressants among all suicides increased from 5.6% to 8.4%. The total suicide rate, however, declined significantly. This was mainly accounted for by the reduced suicide rates by hanging and carbomonoxide poisoning, which outnumbered the increased figures of suicides by poisoning. On the whole, 82% of suicides by antidepressants were committed by use of tricyclics. Use of doxepin and amitriptyline remained steady, and their FTIs were constantly high. The lowest FTIs were associated with fluoxetine, citalopram, mianserin and moclobemide. LIMITATIONS: The method ignores causality between the increased use of SSRIs and suicide mortality. Various factors affecting risk of suicide or choice of a method remain outside the scope of the data. CONCLUSION: The increased use of SSRIs coincided with a significant decline in suicide mortality. However, suicides by use of antidepressants showed an upward trend. Therefore, in suicide prevention, risks and benefits of antidepressants should be considered in choosing treatment for depressive patients. PMID- 9858082 TI - A brain MRI study in subjects with borderline personality disorder. AB - BACKGROUND: There have been only a few brain computed tomography imaging studies, with mostly negative findings, in subjects with borderline personality disorder (BPD). This is the first MRI study which evaluated the structural abnormalities of the brain in subjects with the sole diagnosis of BPD. METHODS: Twenty-five subjects with BPD were compared with age-, gender-matched healthy comparison subjects (n=25) on volumes of the frontal lobes, the temporal lobes, the lateral ventricles, and the cerebral hemispheres in brain magnetic resonance imaging. RESULTS: Subjects with BPD had a significantly smaller frontal lobe compared to comparison subjects (multivariate regression analysis, t=2.225, df=46, P=0.031). There were no significant differences in volumes of the temporal lobes, the lateral ventricles, and the cerebral hemispheres between subjects with and without BPD. LIMITATIONS: Strict inclusion and exclusion criteria employed in the present study may make it difficult to generalize our findings. The gray matter and white matter of the brain were not measured separately. Differences in head tilt during image acquisition were not corrected. CONCLUSIONS: The current study reports a smaller frontal lobe volume on brain MRI in subjects with BPD compared with healthy comparison subjects. This finding may serve as a potentially useful biological variable that may allow for subtyping BPD. PMID- 9858083 TI - Use of topiramate, a new anti-epileptic as a mood stabilizer. AB - RATIONALE: Because some anti-epileptic drugs (AEDs) are effective in bipolar affective disorders, the new AED topiramate (TPM) may be effective in psychiatric illnesses. TPM was evaluated in mood disorders refractory to previous therapies including newer AEDs. METHODS: Charts of 58 consecutive patients, 39 outpatients (15 males, 24 females) and 19 inpatients (6 males, 13 females) were reviewed. TPM 25 mg. b.i.d. was added to existing therapy and titrated in 50 mg increments every 3-7 days. Improvement was rated on a Likert global assessment scale of marked, moderate, mild, or no improvement or worse, based on quality of sleep, appetite, mood, and concentration. RESULTS: Of the 58 patients with psychiatric disorders, 44 patients had rapid cycling bipolar disorders characterized by manic, hypomanic, or mixed episodes. Eighteen patients had previously failed to respond to lamotrigine and/or gabapentin in addition to conventional mood stabilizers. Fourteen were Bipolar I, six Bipolar II, and seven mixed, ten patients had cyclothymic disorder, seven had bipolar disorder not otherwise specified. Of the remaining 14 patients, nine had schizoaffective disorder, three patients had dementia and two had psychosis. Mean duration of TPM treatment was 16.0 weeks; mean TPM dosage approximately 200 mg/day. Thirty-six of 58 (62%) patients exhibited marked or moderate improvement, usually within days or weeks. Twenty-three of 44 (52%) patients with bipolar affective disorders showed marked or moderate improvement. Minimal/no improvement was observed in 16; six were rated as worse. Adverse events included delirium in one patient with Bipolar Disorder Type I who overmedicated with TPM (800 mg) and tranylcypromine sulfate (170 mg) combined with alcohol. Other adverse effects were minor and included: paresthesias, somnolence, fatigue, impaired concentration and memory, nausea, and diarrhoea. LIMITATIONS: This study was performed in a nonrandom open and retrospective fashion. Therefore, any findings are limited by the design of this study. CONCLUSION: TPM may be useful in patients with mood disorders unresponsive to traditional therapy and warrants further clinical investigation. PMID- 9858084 TI - The effect of prophylactic lithium treatment on mortality and suicidal behavior: a review for clinicians. AB - Prophylactic lithium treatment lowers the frequency of recurrences in manic depressive illness. Does it also reduce mortality and suicidal behavior? In this paper all relevant publications were reviewed. The mortality of lithium-treated patients was compared with the mortality in the general population, and the patients' suicidal behavior when they were on lithium was compared with their behavior when they were not. The mortality of manic depressive patients is two to three times higher than that of the general population, a highly significant difference. In the studies reviewed here the patients' mortality during lithium treatment was not significantly higher or only moderately higher than in the general population. After discontinuation of lithium the mortality was again significantly higher. The number of patients attempting suicide was 6-15 times lower and the number of patients completing suicide was 3-17 times lower when they were on lithium than they were not. Similar observations have not been reported for prophylactic treatment with other mood stabilizers. For methodological reasons explained in the introduction, these observations cannot prove that lithium treatment has a mortality-lowering, antisuicidal effect. But they are compatible with such an assumption. It must be the duty of psychiatrists to keep the possibility of an antisuicidal action of lithium in mind when they choose prophylactic treatment for patients with severe depressions or suicidal thoughts or suicide attempts in the past or combinations of these. PMID- 9858085 TI - Suicide mortality in patients on lithium maintenance therapy. AB - Mood disorders are frequently recurrent and it has been shown that maintenance treatment can reduce long-term morbidity in this condition. It has also been shown that mood disorders carry an increased risk of suicide and that a significant proportion of individuals who commit suicide suffer from a mood disorder. This paper reports the results of a long term follow-up of a cohort of patients attending a specialist mood disorder clinic over a period of 18 years. Sixty-seven suffered from unipolar depression and 36 had bipolar or schizo affective disorders In order to qualify for entry to the cohort the unipolar patients had to have had at least three episodes of depression and those with bipolar disorders had to have had at least three episodes - with at least one manic episode and one depressive episode. All patients were treated with lithium. The initial treatment refusal rate and drop our rates were low. The mortality from suicide in this group was compared with that reported in five recent studies - all of which involved patients who had not been given maintenance therapy. The standardised mortality ratio (SMR) for all causes for the whole group was 0.93. There were two suicides. In one case the patient had continued treatment with lithium until death and in the other the patient had discontinued treatment 12 months before death. The overall suicide rate was 1.3 per 1000 patient years. Amongst similar groups of patients who had not been given maintenance therapy suicide rates of about 5.5 per 1000 patient years have been reported. It is concluded that maintenance treatment of mood disorders reduces the suicide rate in this vulnerable group of patients. PMID- 9858086 TI - The psychological autopsy approach to studying suicide: a review of methodological issues. AB - The psychological autopsy approach to studying suicide is becoming an increasingly used research method. It presents considerable methodological problems. In order to assist future researchers in this field and to help readers assess reports of psychological autopsy studies the authors have reviewed these issues on the basis of their own experience and those of other workers. The areas covered include research design, identification of subjects, sources of information and the particular issues concerned with approaching relatives and other informants, choice and recruitment of controls, the difficulties of conducting psychological autopsy interviews with relatives, problems for interviewers, the selection of appropriate measures to obtain information, and achieving valid and reasonably reliable conclusions from diverse information sources. PMID- 9858087 TI - Iowa record-linkage study: death rates in psychiatric patients. AB - The Iowa record-linkage study was developed to investigate death rates in psychiatric patients, and involved computer matching of death certificates with a roster of patients. A list of all patients admitted to our hospital from 1972 through 1981 was obtained and after removing duplicate entries the list was pared to 5412 names. The record included multiple identifiers (e.g., name, gender, date of-birth, hospital number). This information was then linked by computer with all Iowa death certificates for the same period; a total of 331 deaths were identified. Patients were assigned to a single psychiatric diagnostic category based on a computer program that reviewed each patient's clinical diagnoses and picked the one with the highest priority in a hierarchy we had created. Age and sex adjusted mortality tables were constructed, allowing us to compute expected numbers of deaths. Relative risk for premature death was greatest among women, and those under 20 years. Risk was associated with all psychiatric diagnoses and was significantly higher among patients of either gender with an organic mental disorder or schizophrenia; women with acute schizophrenia, depressive neuroses, alcoholism, drug abuse, and psychophysiological disorders; and men with neuroses. Death from natural causes, especially from heart disease, was significantly excessive among women, while death from accidents and suicides was excessive for both men and women. The overall SMR was 1.65 (P < 0.001). Most importantly, we found that the greatest excess of mortality occurred within the first 2 years following hospital discharge. Thus, we were able to demonstrate that risk of mortality in general, and of suicide specifically, differed according to age, gender, diagnosis, and portion of the follow-up. We have subsequently used this method to investigate specific risk factors associated with mortality in mood disorders, schizophrenia, and antisocial personality disorder. Findings from these studies are reported. PMID- 9858088 TI - Rating scales for mania: is self-rating reliable? AB - This commentary addresses the issue of whether self-rating mania scales are reliable. After a brief introduction, several newly developed self-rating mania scales are described and critically reviewed. Among the issues discussed are: the practical utility of self-assessment in patients with mania; the influence of severity of illness on ability to self-report; and, the extent to which the presence of psychosis or the absence of insight significantly interferes with a patients' ability to reliably complete self-rating measures. Data from recent studies suggests that self-rating mania scales are both reliable and valid for patients with manic symptoms, including those with psychotic features and those having little or no insight into their illness. The merits of including self rating measures in the assessment of mania are discussed in terms of their value for both clinical and research purposes. PMID- 9858089 TI - Progress and pitfalls: bipolar molecular linkage studies. AB - This paper reviews the history of molecular genetic linkage studies of bipolar disorder. The topic is introduced with a brief discussion of various genetic concepts, including linkage, lod scores and non-parametric statistics. It is emphasized that criteria for declaring linkage must include independent confirmation by a second group of investigators. Given that the inherited susceptibility for bipolar disorder is most likely explained by multiple genes of small effect, simulations indicate that universal confirmation of valid linkages cannot be expected. With this background, several valid linkages of BP disorder to genomic regions are reviewed. These valid linkages include 18p11, 18q22, 21q21, Xq26 and 4pter. The issue of anticipation and expanding triplet repeats is discussed. Finally, there is a brief section on recommendations for future genetic linkage studies of bipolar disorder. PMID- 9858090 TI - Neural retinal contribution to the slow negative potential of the canine electroretinogram. AB - To establish the normal waveform of the electrical responses from canine eyes, electroretinograms and the light peak were recorded in hybrid and beagle dogs under general anesthesia and artificial ventilation. The neural retinal and retinal pigment epithelial components were pharmacologically isolated by intravitreal glutamate injection and systemic sodium iodate administration, respectively. The a- and b-waves elicited by either flash or rectangular stimuli, the oscillatory potentials elicited by flash stimuli and the light peak elicited either by a single maintained illumination or by repetitive stimuli were almost identical with those of other vertebrates thus far studied. In contrast, in response to rectangular (several-second duration) stimuli, the c-wave was usually absent and was replaced by a slow cornea-negative potential that had a time course similar to that of the c-wave in other species. This slow negative potential was elicited at such low stimulus intensities that the a-wave was absent, was deepened by an intravenous administration of sodium iodate, was not affected by an intravitreal injection of sodium glutamate and was shallowed during the light peak. These results suggest that the slow negative potential that replaces the c-wave consists of large slow PIII, small retinal pigment epithelial c-wave and negligible contribution from the late photoreceptor potential and the dc component of the PII. PMID- 9858091 TI - Effects of experimental scotomata on sequential pattern-onset, pattern-reversal and pattern-offset visual evoked potentials. AB - The effect of experimental scotomata on visual evoked potentials to half-field stimulation using sequential checkerboard onset, reversal and offset was investigated in 10 normal subjects to assess the relative sensitivity of the three stimulus modes, and the contributions of pathways subserving macular and paramacular parts of the visual field. Four scotoma sizes (0-1.5 degrees , 0 degrees - 2 degrees, 0 degrees -3 degrees and 0 degrees -4.5 degrees ) were used to mask the central part of the stimulus field (0 degrees -12 degrees ). Five check sizes (6', 12', 20', 50' and 80') were presented for each scotoma size. Peak-to-peak amplitudes and peak latencies of components on the ipsilateral and contralateral sides of the scalp to the stimulated half-field were measured. Scotoma size was highly significant in influencing component amplitude (p < 0.0001) and latency (p < 0.03) of all ipsilateral and contralateral onset components and all ipsilateral reversal and offset components. Components that were attenuated to the greatest extent with the smallest 0 degrees -1.5 degrees scotoma were the contralateral onset P105 and ipsilateral reversal P100 and N145. Onset CIII, reversal N80, and offset N85 and P110 only showed a significant attenuation after the use of scotomata of 0 degrees -3 degrees and larger. Our results show that scotoma size is a significant factor in influencing all the major visual evoked potential components, with the exception of reversal and offset contralateral potentials (N105 and N115), probably reflecting their paramacular origins. Reversal, ipsilateral P100 and N145, and onset, contralateral P105, appear to be predominantly of macular origin and the most sensitive potentials for detecting effects of small central scotomata. PMID- 9858092 TI - Oscillatory potentials of the electroretinogram in hypertensive patients with different antihypertensive treatment. AB - Oscillatory potentials of the electroretinogram are useful to confirm the diagnosis of essential hypertension. Thirty-five hypertensive patients underwent primary antihypertensive therapy with four different treatments. Oscillatory potentials were recorded before the treatment and after 12 months. The oscillatory response increased in a statistically significant manner in the angiotensin-converting enzyme inhibitor group. This is probably caused by the vasodilatation mechanism, which increases the retinal blood flow. PMID- 9858093 TI - Comparisons of contact lens, foil, fiber and skin electrodes for patterns electroretinograms. AB - Pattern electroretinograms are small physiologic signals that require good patient cooperation and long recording times, particularly when conditions are not optimal. Six electrodes were compared to evaluate their efficacy. Pattern electroretinograms were recorded in eight healthy volunteers to high-contrast, pattern-reversal checks (40' width) with Burian-Allen, DTL fiber, C-glide, gold foil, HK loop and skin electrodes. Raw data for 320 reversals were analyzed off line to evaluate signal amplitude, quality, P50 and N95 peak times, artifact rate and electrical noise. Insertion time, impedance and subjective comfort were also assessed. The Burian-Allen contact lens electrode gave the largest signal and lowest impedance but was the least comfortable and had the highest artifact rate (p < 0.01). A skin electrode on the lower eyelid produced the smallest pattern electroretinogram with the poorest quality (p < 0.05). The four other electrodes were foil or fiber electrodes in contact with the tear film, conjunctiva and/or the inferior cornea. The signal from these showed only minor differences. When electrodes are compared for pattern electroretinograms recording, the foil and fiber electrodes do not differ substantially but contact lens and skin electrodes show substantial disadvantages. PMID- 9858094 TI - Electroretinographic changes in eyes with idiopathic macular hole treated by vitrectomy. AB - We recorded full-field electroretinograms before and after vitrectomy in 18 eyes of 18 patients with idiopathic macular hole. The results were compared between affected and fellow eyes in the preoperative and early (within 10 days) and late (3 to 6 months) postoperative periods. No significant changes between affected and control eyes were found in the amplitude of the rod electroretinogram, mixed cone-rod electroretinogram, cone electroretinogram a- and b-waves or 30-Hz flicker electroretinogram in all stages of the study. The peak implicit times of the cone electroretinogram (a- and b-wave) and dark- and light- adapted oscillatory potential (O1-O4), however, were delayed. Also, the amplitude of the oscillatory potentials (O1+O2+O3+O4) was significantly reduced in the early postoperative period. By the late period, all of these changes had resolved. We concluded that electrophysiologic changes were derived from a transitory dysfunction of the inner retina. The possible causes of the electroretinographic changes would include mechanical trauma of the surgery, intravitreous air tamponade or the changes in vitreous electrolytes after surgery. Oscillatory potentials were more sensitive than a- and b-waves in eliciting dysfunction of the inner retina in operate on eyes. PMID- 9858095 TI - Simultaneous pattern electroretinogram and visual evoked potential recordings in dyslexic children. AB - To help clarify the conflicting evidence of neurophysiologic abnormalities in children with reading problems (dyslexia), we examined pattern electroretinograms and visual evoked potentials to stimulation with checks of 24', 49' and 180', each at 5%, 42% and 100% contrast, in a group of dyslexic children and a group of normal (i.e., normally reading) children. Neurophysiologic difference between the groups was restricted to the visual evoked potential, which showed a significant prolongation of the P100 wave in dyslexic children at the highest contrast (100%) and the smallest checks (24'). There was no significant difference between normal and dyslexic children in the P50 and N95 pattern electroretinogram waves. These results support the assumption of a visual deficit in dyslexic children. However, they are not consistent with the evidence of an isolated deficit of the magnocellular function, which, theoretically, would cause more prominent visual evoked potential changes to lower contrast and the largest check stimuli. PMID- 9858096 TI - The c-wave of the electroretinogram recorded under clinical conditions from rabbits. AB - To determine whether large and repeatable c-waves can be recorded from rabbits with equipment already in use in clinical electroretinographic laboratories, the Burian-Allen electrode, connected bipolarly or monopolarly, was used to record electroretinograms from pigmented rabbits. The Jet electrode was also used. The c waves elicited by long-duration (4-second) stimuli were compared to those elicited by stroboscopic stimuli. In addition, the c-waves recorded with direct coupled amplification were compared to those recorded with condenser-coupled amplification (one-half-amplitude bandpass=0.1 Hz). The b-wave amplitude was not altered by the amplifier coupling or by the two stimulus durations. The largest c waves were elicited by 4-second-duration stimuli and recorded with direct-coupled amplification. Although the c-wave amplitude was reduced by stroboscopic stimuli and by condenser coupling, large and repeatable c-waves were elicited by stroboscopic stimuli and recorded with condenser-coupled amplification. A comparison of stimulus duration and amplifier coupling showed that the stimulus duration was more important in recording large-amplitude c-waves. Similar results were obtained with the Jet electrode. We conclude that repeatable and large c waves can be elicited by a stroboscopic stimuli and can be recorded with condenser-coupled amplification with good low-frequency response from rabbits. PMID- 9858097 TI - Seasonality of birth in epilepsy: a Danish study. AB - OBJECTIVES: A recent British epidemiological study, having found that the seasonality of birth in a large epileptic sample was significantly different from that of the general population, has pointed to neurodevelopmental disruption as the likely mechanism to cause at least part of the epilepsies of unknown aetiology. The aim of this study is to replicate the British study using a large Danish sample. MATERIAL AND METHODS: The population studied is composed by all the 50,886 patients discharged from Danish Hospitals, with a diagnosis of epilepsy, in the period from 1977 to 1993. The seasonal pattern of birth in this sample has been compared, using regression methods, with all the live births in Denmark. RESULTS: The results were strikingly similar to the British study, with a deficit of epileptic births in September and an excess during the winter months. CONCLUSION: This study seems to confirm the seasonal presence in the environment of an aetiological factor(s) for epilepsy which acts in the perinatal period disrupting the neurodevelopment. PMID- 9858098 TI - Multiple sclerosis in North African migrants to France. AB - Among some 7500 respondents with known place of birth who had completed a nationwide questionnaire survey for multiple sclerosis (MS) in France in 1986, there were 260 born in former French North Africa (Algeria, Morocco, Tunisia). They had migrated to France between 1923 and 1986, but 66% came between 1956 and 1964. Two-thirds were from Algeria, where virtually the entire European population had emigrated in 1962 at the end of the Algerian war for independence. The migrants were younger at prevalence day (mean 43.4 years) and at onset (29.4 years) than the French-born MS (46.6; 31.3 years). Eight migrants lacked age information. The 225 migrants with onset more than 1 year after immigration presumably acquired their MS in France. They provided an age adjusted (US 1960) MS prevalence rate 1.54 times that for all France. If the latter is taken at 50 per 100,000 population their estimated adjusted rate is 76.8 with 95% confidence interval of 67.1 to 87.5. The other 27 with presumed acquisition in North Africa gave an estimated adjusted prevalence of 16.6 per 100,000 (95% CI 10.9-24.1). For those migrants with acquisition in France there was a mean interval of 13 years between immigration or age 11 and clinical onset, with a minimum of 3 years. This series provides further support for the theses: 1) that MS is primarily an environmental disease acquired after childhood; 2) that acquisition requires prolonged or repeated exposure (here 3 years for these medium-to-high MS risk migrants) followed by a prolonged latent or incubation period between acquisition and symptom onset (here 10 years); and 3) that this disease is most likely a widespread but unknown persistent infection which results in clinical MS in only a small proportion of those affected. PMID- 9858099 TI - Analysis of lymphocyte subpopulations in cerebrospinal fluid and peripheral blood in patients with multiple sclerosis and inflammatory diseases of the nervous system. AB - We analysed different subsets of lymphocytes from peripheral blood (PB) and cerebrospinal fluid (CSF) by flow cytometry in order to determinate alterations in patients with multiple sclerosis (MS) in acute relapse and viral inflammatory neurological disease (IND). We found increased levels of adhesion molecules (LFA 1 and beta1 integrin) in the CSF of patients with MS and IND compared to NIND. CD4+/CD8+ ratio was significantly higher in CSF of MS as compared with all groups analysed and compared with PB. We detected a significantly higher expression of the interleukin-2 receptor in PB of MS patients when compared with other groups. In patients with IND a significant higher expression of the interleukin-2 receptor was found in the CSF compared with MS and NIND. Our findings indicate that the activation of T lymphocytes primarily occurs in the peripheral immune compartment in MS and the increase of adhesion molecules in CSF is related to inflammatory disorders and not only to MS. PMID- 9858100 TI - Genetic susceptibility to multiple sclerosis in Sardinians: an immunological study. AB - OBJECTIVES: We studied the in vitro production of variably MS-related cytokines from Sardinian MS and healthy donors bearing the two "Sardinian" MS-associated HLA-DR alleles: DR3 and DR4, with the purpose to evidentiate possible differences in their immune response. MATERIALS AND METHODS: ELISA were used for detection of cellular products by mitogen-activated peripheral blood mononuclear cells. RESULTS: PHA-activated HLA-DR4+/DR3- mononuclear cells produce significantly higher amounts of TNF-alpha compared with the DR3+/DR4-. In addition, homozygous HLA-DR3+ mononuclear cells from MS patients produce significantly lower amounts of IL-10 than those from homozygous HLA-DR3+ healthy donors. CONCLUSION: The abnormal production of detrimental or regulatory cytokines may account for the genetic susceptibility to MS in different HLA-subgroups of Sardinian MS patients. PMID- 9858101 TI - Reduced intracortical facilitation in patients with cerebellar degeneration. AB - OBJECTIVES: Transcranial magnetic stimulation (TMS) was used to study intracortical inhibitory and excitatory phenomena in patients with cerebellar ataxia. METHODS: Motor evoked potentials (MEP) following single and paired TMS were recorded from the first dorsal interosseus muscle (FDI) in 15 patients with autosomal-dominant or idiopathic cerebellar ataxia and 15 age matched normal controls. RESULTS: MEP amplitudes after paired TMS with short interstimulus intervals (1-4 ms) showing intracortical inhibition in the control group were not significantly different in the patient group. In contrast, with longer interstimulus intervals (8-20 ms) mean MEP amplitudes were significantly reduced in the patient group, indicating a decrease of intracortical facilitation. The mean postexcitatory inhibition after TMS was also significantly prolonged in the patient group. CONCLUSION: Our findings support the idea that the cerebellum physiologically exerts a facilitatory influence on the motor cortex which is decreased in patients with a cerebellar degeneration. PMID- 9858102 TI - Clinical and molecular findings in the first identified Italian family with dentatorubral-pallidoluysian atrophy. AB - OBJECTIVES: Dentatorubral-pallidoluysian atrophy (DRPLA) is a rare autosomal dominant neurodegenerative disorder mostly observed in Japan, rarely reported in American and North European populations. The aim of this study is to characterize the clinical and molecular features of the first identified Italian DRPLA family. PATIENTS AND METHODS: We describe a 33-year-old female presenting with ataxia, intellectual decline, epilepsy, and choreoathetosis with an adult age onset. Genomic DNA was isolated from peripheral blood lymphocytes of the patient and of her healthy family members following standard procedures. Molecular tests were performed including genetic analysis for SCA1, 2, and 3 (spinocerebellar ataxias), Huntington's disease (HD) and DRPLA, due to a possible overlapping in clinical presentation. RESULTS: Molecular analysis revealed in our patient the presence of a pathological CAG expansion within the DRPLA gene. We have also documented the presence of a smaller CAG expansion in her apparently healthy brother, excluding the possibility of a de novo mutation. CONCLUSION: We conclude that both siblings may have inherited the molecular lesion from their deceased father, the mother being normal at molecular evaluation. Our kindred and a previously reported family from the island of Malta suggest that hereditary DRPLA may also be present in the Mediterranean area. PMID- 9858103 TI - The serial use of two provocative tests in the clinical diagnosis of carpal tunel syndrome. AB - OBJECTIVES: 1) To examine the diagnostic value of the newly proposed manual carpal compression test (mCCT). 2) To evaluate the test efficacy of a serial use of mCCT and Phalen's test (PT) in the clinical diagnosis of carpal tunnel syndrome (CTS) in comparison with standard nerve conduction studies. MATERIALS AND METHODS: Prospective, controlled study of 47 patients with 63 symptomatic CTS hands according to clinical as well as electrophysiological criteria. In the control group, 39 hands of 20 healthy volunteers were examined. Clinical neurological examination including mCCT and PT and standard nerve conduction studies were always performed: sensory nerve conduction velocity third and fifth digit to wrist, median distal motor latency. Calculation of sensitivity, specificity, positive and negative predictive value for the mCCT, PT and mCCT+PT, i.e. at least one of both tests positive after sequential application. RESULTS: Sensitivity 83% for mCCT, 79% for PT and 92% for mCCT+PT. Specificity 92% for all three conditions. Excellent positive predictive value of 95% and good negative predictive value of 88% for mCCT+PT. CONCLUSION: The serial application of mCCT and PT is very useful in the clinical diagnosis of CTS. This finding facilitates the clinical diagnosis by the primary physician, it helps to improve referral decisions to electrophysiological departments and other ancillary examinations and may reduce costs. PMID- 9858104 TI - Neurophysiological and anatomical variability of the greater auricular nerve. AB - OBJECTIVE: To study the normal variation in greater auricular nerve (GAN) conduction and anatomical course. MATERIALS AND METHODS: GAN nerve conduction was studied in 77 healthy adults while 17 GANs were dissected in 10 cadavers. RESULTS: The largest sensory nerve action potentials (SNAP) were recorded when the stimulator was placed 6 or 7 cm from the external acoustic meatus (EAM). Amplitude decreased significantly with age (P = 0.001). The dissected GANs entered the subcutaneous layer between 55 and 105 mm (median 68 mm) from EAM. The postauricular branch was found in 13 of 17 cases. CONCLUSION: A large variation was observed both anatomically and by way of electrical stimulation. SNAP variation does probably reflect the anatomical variation in GAN course, branching, and terminal distribution. The best sites for stimulation were found 6 and 7 cm from EAM at the posterior sternocleiodomastoid border. The maximal amplitude, right:left amplitude ratio, and distal latency may be useful variables in GAN conduction studies. PMID- 9858105 TI - Clinical manifestations of 20 Taiwanese patients with paroxysmal kinesigenic dyskinesia. AB - INTRODUCTION: We compared the clinical manifestations and response to medications between familial and sporadic patients with paroxysmal kinesigenic dyskinesia (PKD), and also between patients with autosomal dominant (AD) and autosomal recessive (AR) inheritance. MATERIAL AND METHODS: This retrospective cohort study included 9 familial and 11 sporadic Taiwanese patients with PKD diagnosed during a 10-year period at one of two hospitals. The mean duration of follow-up was 3.8 +/- 2.7 years. Each patient was interviewed and their medical records, as well as videotape recordings of PKD attacks in 6 patients, were used for analysis. Patients were treated with either carbamazepine or phenytoin, and the efficacy of sodium valproate was tested in 5 patients. RESULTS: No single distinguishing feature in terms of clinical manifestations or therapeutic response was found to differentiate among familial, and sporadic cases, or between AD and AR inheritance. Carbamazepine and phenytoin were superior to sodium valproate in treating both familial and sporadic PKD patients, and both drugs resulted in almost complete remission of attacks. CONCLUSION: Our findings indicate that the sporadic and familiar forms of PKD, as well as the AR and AD inherited types, are similar in terms of clinical manifestations and response to treatment. The functional status and prognosis of our Taiwanese patients suggest that PKD is a relatively benign entity. PMID- 9858106 TI - Diabetic neuropathy in elderly Type 2 diabetic patients: effects of insulin treatment. AB - OBJECTIVES: To study the occurrence of diabetic neuropathy and the effect of insulin treatment in elderly Type 2 diabetic patients. MATERIAL AND METHODS: In 38 patients and 20 controls symptoms and neurophysiological examinations including electroneurography, vibration perception and temperature discrimination thresholds were investigated. Patients were randomized to insulin (n = 18) or sulfonylurea (n = 16) treatment and were re-investigated after 1 year. RESULTS: Neuropathy was present in 21/38 patients (56%). It was asymptomatic in 17/38 (45%) and symptomatic in 4/38 (11%). The occurrence of neuropathy was less common in healthy controls, 3/20 (15%) (P < 0.01). Temperature discrimination thresholds was the test that most often revealed pathology. The metabolic control after 1 year was significantly improved in the insulin treated group and unchanged in the sulfonylurea treated group. There were no changes as regards occurrence of neuropathy between or within the two treatment groups after 1 year. CONCLUSION: Diabetic neuropathy is common among elderly Type 2 diabetic patients. It is mostly asymptomatic. Improvement was not seen after 1 year of insulin treatment. PMID- 9858107 TI - Effects of intra-arterial papaverine on the chronic period of cerebral arterial vasospasm in rats. AB - OBJECTIVES: The effect of intra-arterial papaverine (IAP) on the basilar artery (BA) and cerebral perfusion pressure (CPP) during the chronic period of the cerebral arterial vasospasm in rats was investigated. MATERIAL AND METHODS: The study was carried out on male, Swiss-Albino rats, the weight of each varied between 200-340 g. A large volume (0.3 cc) of nonheparinized, autolog blood was utilized in order to cause a subarachnoid haemorrhage. For the measurement of the changes in BA diameter, the angiograms were made prior to the subarachnoid haemorrhage, 48 h after the subarachnoid haemorrhage, and in 1, 15, 30, and 60 min after papaverine infusion into the vertebral artery. The BA vascular index was found separately for each angiogram. At each stage of the procedure mean arterial blood pressure (MAP) and intracranial pressure (ICP) were monitored. RESULTS: BA diameter measurements were found to be 226+/-32 microm in pre haemorrhage angiograms and 145+/-44 microm in angiograms 48 h after the subarachnoid haemorrhage. In the angiograms immediately after IAP, it was found that the BA diameter reached about 92% (206+/-41 microm) of control values. But, in the angiograms 15 min after IAP, it was observed that BA underwent a spasm again. CONCLUSION: The dilatator effect of IAP on BA was temporary. Additionally, in the chronic vasospasm period when cerebral autoregulation mechanisms are impaired and CPP decreased significantly, IAP has adversely affected CPP decreasing MAP. PMID- 9858108 TI - A reversible neuronal antibody (anti-Tr) associated paraneoplastic cerebellar degeneration in Hodgkin's disease. AB - Paraneoplastic cerebellar degeneration (PCD) has been associated with a variety of neoplasms, most commonly with gynecologic tumors, breast cancer, small cell lung cancer, and Hodgkin's disease (HD). In some patients PCD is associated with circulating antineuronal antibodies like anti-Hu, anti-Yo or anti-Ri. Previously, only 5 patients with a new antineuronal antibody called anti-Tr, proposed to be specific for HD, have been reported. We describe 1 further patient with HD and reversible PCD with a decline in anti-Tr antibody titers in cerebrospinal fluid and serum corresponding to the improvement of clinical symptoms. At the present time the immunoreactive pattern observed in rat cerebellum is the only way to identify anti-Tr antibodies and differentiate them from other antibodies that immunoreact with the Purkinje cells. PMID- 9858109 TI - Myophosphorylase deficiency and limb-girdle muscular dystrophy in the same pedigree. AB - We report 2 familial patients with limb-girdle muscular dystrophy (LGD). The parents of patient 1 showed a consanguineous marriage and patient 2 was a paternal cousin of patient 1. Slowly progressive muscular weakness/wasting and dystrophic changes in the biopsied muscles were observed in both patients. However, a quantitative assay revealed a severely reduced myophosphorylase activity in patient 1 with normal activity in patient 2. A semi-ischemic exercise test disclosed no elevation of venous lactate in patient 1 with a normal increase in patient 2. A leukocytes DNA analysis in patient 1 did not show the gene deficits previously recognized in patients with McArdle's disease (McD). Patient 1 may only have abnormal myophosphorylase activity with dystrophic changes secondary to the myophosphorylase deficiency or coincidentally two genomic abnormalities for McD and LGD. LGD still has heterogenous etiologies and the responsible genes for these two disorders may be closely mapped. PMID- 9858110 TI - Measuring blood pressure in children and adolescents. PMID- 9858111 TI - Isolated systolic hypertension: an important cardiovascular risk factor. AB - Hypertension is an established risk factor for cardiovascular disease morbidity and mortality. Randomized trials of antihypertensive therapy have demonstrated the benefits of treating diastolic blood pressure, and recently the value of treating isolated systolic blood pressure has also been established. There is an excess risk of cardiovascular disease in subjects with borderline isolated systolic hypertension. In fact, data from men screened for the Multiple Risk Factor Intervention Trial show that the great majority of excess deaths are in those with high-normal systolic blood pressure or with stage 1 hypertension, i.e., systolic blood pressure 130 to 159 mmHg. Similarly, data from the Framingham Heart Study and the Physicians' Health Study emphasize the importance of mild elevations of systolic blood pressure. As age increases the hemodynamic patterns of blood pressure change due to an increase in large artery stiffness, and borderline isolated systolic hypertension becomes the dominant form of hypertension. These facts make the prevention and control of borderline isolated systolic hypertension a key strategic challenge in the effort to prevent excess mortality attributable to blood pressure levels above normal. PMID- 9858112 TI - No impaired cognitive function in treated patients with mild-moderate hypertension compared to normotensive controls. AB - Hypertension is a predictor for impaired cognitive function and dementia in several prospective studies. It is currently under debate whether treatment of hypertension, and thus blood pressure lowering, is another risk factor for cognitive decline. We recruited a sample of 123 treated hypertensive patients and 76 normotensive controls, from a population-based study in primary health care, for screening of blood pressure, metabolic variables and cognitive function, as measured by the Mini-Mental State Examination (MMSE). Treated hypertensives had higher blood pressure but did not differ in cognitive function from the normotensives. Neither educational level nor metabolic variables confounded the findings. In conclusion, treated hypertensives did not differ in cognitive function from normotensive controls. This does not support the notion that pharmacological blood pressure reduction impairs cognitive function. PMID- 9858113 TI - Muscle sympathetic vasoconstrictor activity in hydrocortisone-induced hypertension in humans. AB - AIM: This study was undertaken to test the hypothesis that increased sympathetic vasomotor drive is responsible for cortisol-induced hypertension. METHODS: Ten healthy male subjects on a fixed sodium diet (150 mmol/day) were randomized to five days of treatment with cortisol (200 mg/day) or placebo in a double-blind crossover study. On day 5 of each treatment, multi-unit muscle sympathetic activity was recorded from the common peroneal nerve. Resting muscle sympathetic activity (MSA) was measured in the recumbent position and stimulated MSA was measured in the final 20 sec of end-inspiratory capacity apnoea and end expiratory apnoea and in the second minute of a cold pressor stimulus. A subgroup of six subjects also underwent identical MSA measurements following 5 days treatment with dexamethasone (3 mg/day). MAJOR FINDINGS: Cortisol, but not placebo, significantly increased systolic (115+/-2 vs 129+/-3 mmHg precortisol vs cortisol day 5, p < 0.001) and diastolic blood pressure (53+/-3 vs 61+/-3, p < 0.05). Resting MSA was significantly reduced by cortisol (23.9+/-2.3 to 5.0+/-2.0 bursts/min, placebo vs cortisol, p < 0.01). Cortisol significantly attenuated the increase in MSA observed at end-inspiratory apnoea (56.3+/-3.9 vs 35.4+/-6.6, p < 0.05) and end-expiratory apnoea (50.5+/-3.5 vs 26.3+/-6.2 bursts/min, n = 8, p < 0.05), and during the cold pressor response (55.0+/-12.7 vs 21.4+/-7.6, n = 5, p < 0.05). Dexamethasone significantly increased systolic blood pressure and suppressed resting and stimulated MSA. No changes in body weight, haematocrit or angiotensin II concentrations occurred during dexamethasone treatment. CONCLUSION: MSA is significantly suppressed by cortisol treatment. As suppression of MSA is also observed during treatment with the pure glucocorticoid dexamethasone, suppressed MSA cannot be attributed to increased plasma volume or to changes in angiotensin II concentration. We conclude that cortisol-induced hypertension is not due to increased muscle sympathetic vasomotor drive. PMID- 9858114 TI - Factors influencing left ventricular mass in salt sensitive and salt resistant essential hypertensive patients. AB - To investigate whether salt sensitivity is associated with differences in left ventricular mass or geometry, salt sensitivity testing and Doppler echocardiography was performed in 30 essential hypertensive patients (7 women and 23 men) with mean age 43+/-9 years. Salt sensitivity was defined as a 10% increase or more in 24-h blood pressure (24hBP) when going from low to high dietary sodium intake based on a single test. Eight patients were characterized as salt sensitive and 22 as salt resistant. At baseline, there was no difference in casual blood pressure (156/103+/-17/9 vs 158/100+/-18/11 mmHg) or 24hBP (152/ 90+/-25/15 vs 159/89+/-19/8 mmHg), in duration of hypertension (5+/-4 vs 4+/-3 years), daily sodium excretion (144+/-68 vs 171+/-68 mmol), left ventricular mass (LVM) (212+/-45 vs 246+/-52 g) or left ventricular relative wall thickness (RWT) between the salt sensitive and salt resistant groups of patients. In the total study population, increased RWT was found in 17 patients, and increased LVM in 10 patients. In only 10 patients were both these variables normal. Left ventricular geometric pattern did not differ between the salt sensitive and salt resistant groups. LVM and RWT were significantly correlated with 24hBP (r = 0.57 and 0.51, respectively; both p < 0.01). Significant correlation was also found between LVM and casual blood pressure, blood volume, body surface area, serum creatinine and albuminuria (r = 0.53, 0.60, 0.54, 0.54 and 0.43, respectively; all p < 0.01). In multiple regression analysis, 24hBP and blood volume were identified as independent predictors of LVM (R = 0.51, p < 0.001). CONCLUSIONS: increased RWT or LVM is common in both salt sensitive and salt resistant essential hypertensive patients. Salt sensitivity status based on a single test does not influence left ventricular hypertrophy or geometry. Twenty-four-hour blood pressure is related to increased RWT and LVM. PMID- 9858115 TI - Performance of a new blood pressure monitor in children and young adults: the difficulties in clinical validation. AB - The aim of this study was to assess the performance of a new ambulatory blood pressure monitor (Dynapulse 5000A) in children and young adults. The reference methods, i.e. the mercury-Doppler (Hg-D) method of blood-pressure measurement (recommended for less than 5-year-old children by the British Hypertension Society criteria) and the mercury-auscultation (Hg-A) technique (recommended for 5 years and above) were initially compared utilizing 132 concomitant systolic blood pressure (SBP) readings taken among 44 subjects (3 in each) aged 2-30 years. Hg-D over-read SBP above that of the conventional Hg-A by a mean of 5.25 mmHg (95% confidence interval (CI) 4.11, 6.4). This difference in measurement between the two techniques was also significantly (p = 0.01) related to age. [Difference in mmHg = 7.83 (95% CI 5.53, 10.12) -0.30 (95% CI -0.54, -0.07) age in years.] The Dynapulse 5000A was therefore validated only against Hg-D throughout the age range and was undertaken utilizing 468 concomitant resting SBP measurements taken among 59 subjects (supine or sitting) using a 'T' system. The Dynapulse was found to give higher SBP readings (average 13.5 mmHg) than Hg-D sphygmomanometry. Although the differences were not dependent on the age of the child, the pulse rate or the body mass index, there was a significant drift in the differences with time (-0.05 mmHg/day). Repeatability of each of the measurement methods calculated utilizing two or three consecutive recordings taken at 1-min intervals showed that variability between repeated measurements made with the Dynapulse (14.8%) was greater than for Hg-D. Repeatability coefficient of the Hg-D (11.2%) was also less than ideal for a reference method. We conclude that Dynapulse 5000A over-reads SBP above that detected by Hg-D, but cannot be fairly graded by British Hypertension Society criteria due to the inconsistency and poor repeatability of the recommended reference methods. PMID- 9858116 TI - Influence of body size and left ventricular ejection dynamics on total arterial compliance determined using Doppler echocardiography and subclavian artery pulse tracings in healthy humans. AB - The aim was to investigate determinants of total arterial compliance in healthy humans. Estimates of aortic root pressures and flow were obtained non-invasively with the calibrated subclavian artery pulse tracing and Doppler echocardiography in 37 males (27-76 years) and 45 females (20-77 years). Total arterial compliance, estimated using a three-element vascular model, correlated positively with body height (r = 0.45, p < 0.01) and acceleration time of aortic root flow (r = 0.32, p < 0.01) and inversely with age (r = -0.34, p < 0.05), heart rate (r = -0.33, p < 0.01), and mean arterial pressure (r = -0.51, p < 0.01). Multivariate analysis indicated that height and heart rate contributed most to the prediction of total arterial compliance. The inclusion of mean arterial pressure within the model significantly reduced the contribution of age, but not that of body height and heart rate. After adjustment for height and heart rate, total arterial compliance did not differ significantly between gender. Thus, total arterial compliance, as assessed in this study, seems to reflect both arterial capacity and viscoelastic properties of the arterial wall. Differences in body size, heart rate and mean arterial pressure should be considered when comparing total arterial compliance in different groups. PMID- 9858117 TI - Enhanced angiotensin II stimulates renal disorders in transgenic Tsukuba hypertensive mice. AB - Tsukuba hypertensive mice (THMs) are transgenic mice carrying human renin angiotensin system (RAS) genes. The aim of this study is to evaluate whether renal disorders are present in THMs. Twenty-week-old THMs and C57BL/6 mice (C57s) were used for this study. Each group consisted of 8 mice. Systolic blood pressure, urinary volume, water intake and urinary albumin excretion were measured in each mouse. Each mouse was then euthanized, and the renal glomerulosclerosis index and glomerular size were measured. Systolic blood pressure of THMs was about 40 mmHg higher than that of C57s. Urinary volume, water intake and urinary albumin excretion were significantly higher in THMs than in C57s. The renal glomerulosclerosis index and glomerular size were also significantly higher in THMs than in C57s. These results suggested that an enhanced renin-angiotensin system, including its hypertensive effects, stimulates albuminuria, renal glomerulosclerosis and glomerular hypertrophy in THMs. PMID- 9858118 TI - Candesartan cilexetil vs losartan. PMID- 9858119 TI - 3-D characterization of ganglion cells of the terminal nerve plexus of rat atrioventricular junction. AB - Three-dimensional (3-D) morphology of neurons of the terminal nerve plexus of the atrioventricular junction was examined in a scanning electron microscope. Distributions of different cell types encountered as well as their relations to different structures of the atrioventricular specialized tissue were also studied. Most neurons were found disseminated in a thin connective tissue layer separating different segments of the atrioventricular conductive tissue from the interventricular septum. Sometimes, they formed small pluricellular ganglia (up to 5 neurons) but, frequently, they occurred isolated in the terminal ramifications of the intramural nerve plexus of specialized tissue. Some intranodal neurons could also be identified. According to their 3-D morphology, nerve cells of the perinodal ganglionated plexus could be divided into three categories: (1) Large unipolar neurons were scattered throughout the atrioventricular junction. Their long and thin axonal projections were often directed towards the interventricular septum. (2) Large pseudounipolar or bipolar neurons were located at a few specific loci, namely all along the bundle of His and its bifurcation into the right and left bundle branches. Frequently, they occurred solitary and immersed amongst strands of surrounding muscle cells. Only occasional synaptic impacts could be identified on the surface of neuronal bodies of these bipolar neurons. On the other hand, their dendritic varicosities were richly innervated. Due to their irregular shape, intimate association with muscular elements and their topographical superposition with occasional spindle like structures, these nerve cells recall prospective sensory neurons involved in integration of mechanical and neural stimuli to the heart. (3) Small multipolar interneurons could be identified in the retronodal ganglion and within right and left bundle branches. The present description of morphological heterogeneity of intramural nerve cells agrees with recent morphological and functional classifications of autonomic neurons and supports the idea that, at the level of the atrioventricular junction, a self-governed neuronal network may be operating. PMID- 9858120 TI - Distribution of neurons innervating the uterus of the pig. AB - The distribution of neurons innervating the uterus of the pig was studied with the use of fluorescent retrograde tracer Fast Blue. Tracer injections were made into the uterine cervix, pericervical, middle and perioviductal part of the right uterine horn. After tracer injection into the uterine cervix tracer-positive neurons were found bilaterally in the inferior mesenteric ganglia, paracervical ganglia, paravertebral ganglia L1-S3 and dorsal root ganglia Th10-L4 and S2-S3. After tracer injection into the pericervical part of the right uterine horn the distribution of tracer-positive neurons resembled the one described earlier, except the tracer-positive neurons were absent in the left dorsal root ganglia. After tracer injection into the middle part of the right uterine horn tracer positive neurons were found in the ipsilateral inferior mesenteric ganglion, bilaterally in the paracervical ganglia and in the ipsilateral paravertebral ganglia (L2-L6). Injection of the tracer into the perioviductal part of the right uterine horn revealed tracer-positive neurons bilaterally in the inferior mesenteric ganglia and paracervical ganglia and single cells in the ipsilateral paravertebral ganglia. PMID- 9858121 TI - Central histaminergic neurons regulate rabbit tracheal tension through the cervical sympathetic nerve. AB - We previously showed that stimulation of the posterior hypothalamus decreases tracheal tension and involves central histaminergic neurons. In the present study, we reveal that central histaminergic neurons project to the rostral ventrolateral medulla and affect cervical sympathetic nervous activity in rabbits. Administration of histamine into the fourth ventricle increased cervical sympathetic nervous activity and decreased tracheal tension. These effects were inhibited by administration of a histamine H receptor antagonist, pyrilamine, into the fourth ventricle. Unilateral injection of DL-homocysteic acid into the tuberomammillary nucleus increased cervical sympathetic nervous activity, an effect was antagonized by bilateral injection of pyrilamine into the rostral ventrolateral medulla. The pulse correlogram between the stimulation pulse applied to the tuberomammillary nucleus and the cervical sympathetic nerve activity showed a mode at 150 to 200 ms, which was reduced by pyrilamine administration into the fourth ventricle. Fibers anterogradely labeled by Phaseolus vulgaris leucoagglutinin (PHA-L) injected into the tuberomammillary nucleus were distributed in the A1, A2, C1, and C2 areas which are determined by tyrosine hydroxylase-immunohistochemistry. PHA-L positive neurons were in close contact with tyrosine hydroxylase-immunoreactive neurons in these four areas. Cell bodies in the tuberomammillary nucleus retrogradely labeled with fluorogold from the rostral ventrolateral medulla were immunoreactive with histamine. These results suggest that an excitatory efferent pathway projects from the tuberomammillary nucleus to the cervical sympathetic nerve and that the histaminergic neurons of this pathway influence tracheal tension through the rostral ventrolateral medulla. PMID- 9858122 TI - Inhibitory innervation of the guinea-pig urethra; roles of CO, NO and VIP. AB - The inhibitory innervation of guinea-pig urethral smooth muscle was investigated histochemically and functionally. The distribution of immunoreactivities to haem oxygenases (HO), neuronal NO synthase (nNOS), and vasoactive intestinal polypeptide (VIP) was studied, and the functional effects of the corresponding putative transmitters, CO, NO, and VIP, were assessed. HO-2 immunoreactivity was found in all nerve cell bodies of intramural ganglia, localized between smooth muscle bundles in the detrusor, bladder base and proximal urethra. About 70% of the ganglionic cell bodies were also NOS-immunoreactive (IR), whereas a minor part was VIP-IR. Some ganglion cells exhibiting tyrosine hydroxylase (TH) activity were demonstrated. Rich numbers of NOS-IR varicose nerve terminals could be found innervating the smooth muscle of the urethra, whereas VIP-IR terminals were less numerous. A rich number of TH-IR terminals were observed. The bladder showed a similar distribution of nerves, although only a few number of TH-IR nerves could be found. In bladder preparations exposed to sodium nitroprusside, cGMP-IR cells could be seen, forming an interconnecting network with long spindle shaped processes. The cGMP-IR cells were especially abundant in the outer smooth muscle layers of the bladder, but less numerous in the urethra. In urethral strip preparations, electrical field stimulation evoked long-lasting frequency dependent relaxations. The relaxations were not inhibited by the NO-synthesis inhibitor, L-NOARG, or enhanced by the NO-precursor, L-arginine. The haem precursor, 5-aminolevulinic acid (5-ALA), or the inhibitor of guanylate cyclase, ODQ, did not affect the urethral relaxations. Exogenously applied NO, SIN-1, and VIP relaxed the preparations by approximately 50%, whereas the relaxation evoked by exogenous CO was minor. These results suggest that CO probably is not involved in non-adrenergic, non-cholinergic inhibitory control of the guinea-pig urethra, where a non-NO/cGMP mediated relaxation seems to be predominant. PMID- 9858123 TI - Involvement of the ipsilateral rostral ventrolateral medulla in the pressor response to L-glutamate microinjection into the nucleus tractus solitarii of awake rats. AB - Microinjection of L-glutamate into the lateral commissural nucleus tractus solitarii (NTS) of unanesthetized rats evokes increases in mean arterial pressure (MAP) and a bradycardia. In a previous study we verified that this increase in MAP is mediated sympathetically because prazosin (i.v.) blocks this response. The aim of the present study was to evaluate the role of the rostral ventrolateral medulla (RVLM) in the pressor response produced by L-glutamate microinjected into the NTS of unanesthetized rats. L-Glutamate was microinjected into the NTS before and 15 and 90 min after microinjection of kynurenic acid into the ipsilateral RVLM. Pressor (+24+/-3 vs. +6+/-3 mm Hg) and bradycardic (-101+/-10 vs. -3+/-12 bpm) responses to L-glutamate microinjected into the NTS (n = 8) were almost abolished 15 min after microinjection of kynurenic acid into the RVLM when compared with control responses. Both pressor (+23+/-6 mm Hg) and bradycardic ( 93+/-16 bpm) responses to L-glutamate into the NTS returned to control values 90 min after microinjection of kynurenic acid into the RVLM. These data indicate that the pressor response to L-glutamate into the NTS is essentially dependent on the ipsilateral RVLM and also that this sympatho-excitatory response is mediated by excitatory amino acid receptors in RVLM neurons. PMID- 9858125 TI - Effect of hibernation on responses of hamster vas deferens to sympathetic nerve stimulation and exogenous neurotransmitters. AB - The present study investigated the responses of the vas deferens to sympathetic nerve stimulation and exogenous neurotransmitters taken from golden hamsters which had undergone 8 weeks of hibernation, 2 h of arousal from hibernation, those exposed to the cold but which failed to hibernate and age-matched control animals. Electrical field stimulation (EFS) of the vas deferens from each group produced frequency-dependent, tetrodotoxin-sensitive contractions. Contractions elicited by low frequencies of EFS in the hibernating group were significantly greater than in the other groups in the absence of any blocking agents. In the presence of the alpha1-adrenoceptor antagonist prazosin (3 microM) responses from all groups were reduced by approximately 40%, with the residual responses from the hibernating group being somewhat increased compared to the other groups. In the presence of the P2X receptor antagonist pyridoxalphosphate-6-azophenyl-2'4' disulphonic acid (30 microM), there was no significant difference in responses from all 4 groups. Exogenously applied beta, gamma-methylene ATP (beta,gamma meATP; 0.1-300 microM), a P2X receptor agonist, and noradrenaline (NA; 30 nM(-1) mM) both caused transient concentration-dependent contractions in all groups of animals. Contractions to beta,gamma-meATP at concentrations above 0.3 microM, and NA above 0.3 microM in the hibernating animals were statistically significantly greater than the cold- and age-matched control groups, although not significantly different from the aroused group. This study has shown postjunctional increases in responses to beta,gamma-meATP and NA as a result of hibernation, possible explanations for these increases are discussed. PMID- 9858124 TI - Diurnal rhythms in norepinephrine and acetylcholine synthesis of sympathetic ganglia, heart and adrenals of aging rats: effect of melatonin. AB - The effect of aging and melatonin on 24-h rhythms in tyrosine hydroxylase activity and 3H - choline conversion into 3H - acetylcholine were examined in cervical, stellate, coeliac-mesenteric and hypogastric ganglia, and in the adrenal medulla and heart of rats. Young (50 days old) and old (18 months old) rats received evening injections of 10 or 100 microg of melatonin or its vehicle for 17 days. In superior cervical, stellate and coeliac-superior mesenteric ganglia, as well as in the adrenal medulla, norepinephrine and acetylcholine synthesis attained maximal values at night (c.a. 2030-0100 h). In the hypogastric ganglion, maximal tyrosine hydroxylase activity occurred at night at both studied ages. Two maxima in acetylcholine synthesis were detected in hypogastric ganglion of young rats (c.a. 1300 h and 0100 h, respectively) while in old rats a single maximum was observed at noon. Cardiac tyrosine hydroxylase activity peaked at early night (c.a. 2200-2300 h) while cardiac acetylcholine synthesis peaked at the afternoon (c.a. 1700-1900 h). Old rats exhibited a significant decrease of rhythm amplitude and increase of mean values in tyrosine hydroxylase activity in autonomic ganglia and adrenal medulla, and abolition of tyrosine hydroxylase rhythm in the heart. Twenty-four hour rhythmicity in acetylcholine synthesis was impaired or abolished in aged rats. Treatment of old rats with 10 or 100 microg melatonin generally augmented amplitude of rhythms and reinduced the nocturnal peak of acetylcholine synthesis in the hypogastric ganglion. Only the high melatonin dose significantly augmented rhythm amplitude of tyrosine hydroxylase activity (superior cervical and coeliac-superior mesenteric ganglia) and acetylcholine synthesis (superior cervical, stellate and coeliac-superior mesenteric ganglia) in young rats. The results indicate that the activity of the central oscillator, driven to the organs in part via the autonomic nervous system, deteriorates significantly with aging and that melatonin may restore partially such a deterioration. PMID- 9858126 TI - Effect of growth hormone-releasing peptide 1-6 on GH secretion-stimulated by GHRH and pyridostigmine in lambs. AB - The GHRP-6 seems to act at a pituitary site, activating different intracellular messenger pathways from those utilized by GHRH, and at the hypothalamic level where receptors for GHRP-6 have been demonstrated. This study examines the effect of GHRP-6 on GH secretion in vitro and in vivo. Lamb adenohypophysial cell cultures were subjected to a challenge with 1) 10 nM GHRH-1-29; 2) 1 microM GHRP 6; and 3) 10 nM GHRH plus 1 microM GHRP-6. Both peptides released GH, GHRP-6 being less potent than GHRH, and the GH response to GHRH and GHRP-6 not being synergistic, without statistically significant differences between GHRH+GHRP-6 and GHRH alone. In in vivo studies, six lambs received 15 microg/kg GHRH (1-29) or 15 microg/kg GHRH plus 10 microg/kg GHRP-6 and six other lambs received 100 microg/kg GHRP-6 or 3 mg/kg pyridostigmine plus 100 microg/kg GHRP-6. The results have shown that the combination of GHRH plus GHRP-6, at low doses, causes higher GH peak (p < 0.05) and higher GH area under curve (p < 0.05) with respect to administration of GHRH alone. The administration of GHRP-6 plus pyridostigmine produced in a stronger GH response to GHRP-6 (100 microg/kg) in the amplitude of the GH peak (p < 0.001) and in the area under the GH response curve (p < 0.001). The complementary interactions of GHRP-6 with GHRH or pyridostigmine in releasing GH seem to indicate independent actions of these compounds. These results suggest that GHRP-6 potentiates the GH secretion stimulated by GHRH at the hypothalamic level, in these animals. PMID- 9858127 TI - Inhibition by estrogens of the oxidant-mediated mobilization of arachidonic acid in hepatocytes. AB - Oxidative stress is associated with alterations in arachidonic acid (AA) metabolism. The present work was performed to assess the effect of the oxidant tert-butyl hydroperoxide on the release of AA from rat hepatocytes, and the possible preventive actions of estrogens on this effect. The exposure of [14C] prelabeled cells to tertbutyl hydroperoxide produced the mobilization of [14C]-AA from hepatocyte lipids, both an intracellular [14C]-AA accumulation and an increased release of [14C]-products into the medium being observed. The formation of lysophospholipids was also enhanced significantly in the presence of the oxidant, thus suggesting the involvement of phospholipase A2 (E.C. 3.1.1.4) in the hepatocyte response to tert-butyl hydroperoxide. Estradiol and 2 hydroxyestradiol (25-100 microM) added in vitro to cell suspensions prevented significantly the oxidant- mediated stimulation of AA release, this effect probably being caused by the estrogen inhibitory actions against cellular lipid peroxidation. PMID- 9858128 TI - Human red cells from prenatal hemopoiesis. Sodium/lithium exchange symmetry. AB - Human red blood cells from prenatal stages of hemopoiesis (umbilical cord blood) and with increased lithium content were submitted to media with varying sodium concentrations in order to characterize extracellular sodium dependent-lithium efflux. On the other hand cells with different sodium contents were submitted to lithium media in order to characterize cellular sodium dependent-lithium influx through the above system. Experimental data were fitted by simple Michaelis Menten kinetics. The Km value for lithium efflux (extracellular sodium-dependent) was significantly lower than the Km value for lithium influx (cellular sodium dependent). The Vmax value for lithium influx was significantly greater than that for lithium efflux. The Vmax/Km ratio values were independent from the way in which the fluxes were measured. It is concluded that the simple carrier cannot be excluded as a model of the Na+-Li+ exchange system in these cells. PMID- 9858129 TI - Effect of RU486 injected on proestrous morning on LHRH, LH and 17beta-estradiol secretion during the estrous cycle in rat. AB - The antiprogesterone RU486 injected on proestrus to cyclic rats advances the preovulatory surge of LH, resulting in a 3-day estrous cycle. To ascertain whether proestrous progesterone secretion regulates ovulation by synchronizing the functions of the ovary, the pituitary and the hypothalamus, in this study the effects of RU486 (4 mg/0.2 ml oil/s.c.) at 09:00 h on proestrus (day 1), on LHRH secretion into the pituitary stalk blood vessels and on peripheral plasma concentrations of LH and 17beta-estradiol at 10:00 h and at 18:00 h on each day of the estrous cycle, have been investigated. Control rats present the expected surges of LHRH and LH at 18:00 h on day 1 and a second increase in LHRH but no LH secretion was found in the afternoon of day 2. RU486 decreases both LHRH and LH surges in the afternoon of day 1, while it increases plasma concentrations of LHRH and LH in the morning of day 2. During the rest of the estrous cycle RU486 injected rats show high basal secretion of LH in comparison to controls, and at 18:00 h of day 4 advanced preovulatory surges of both LHRH and LH are present in RU486-injected rats. In relation to 17beta-estradiol concentrations, RU486 injection increases those on day 3 and induces an advanced preovulatory surge of 17beta-estradiol in the morning of day 4. These results show that, in the 4-day cyclic rat, the 24 hours-shortening of the estrous cycle induced by the antiprogesterone RU486 is due to the enhanced secretion of estrogen during the days of metestrus and diestrus, which stimulates the release of a surge of LHRH in the afternoon of diestrus and this, in turn, induces the advanced preovulatory surge of LH. PMID- 9858130 TI - Effects of orange and apple pectin on cholesterol concentration in serum, liver and faeces. AB - To investigate the effects of pectin on cholesterol metabolism, normal rats were fed for three weeks a diet containing 2.5 or 5 % apple or orange pectin, or without pectin (control). Cholesterol concentrations were determined in faeces after 1, 2 and 3 weeks of treatment, and in liver and serum at the end of the experimental trials. Cholesterol concentration in faeces showed a significant increase by week 3 in rats fed 5 % orange or apple pectin. Hepatic cholesterol concentration declined significantly in all pectin-fed groups. Serum cholesterol only declined significantly in apple-fed groups. The decrease of cholesterol levels in liver and serum, and its increase in faeces could explain the beneficial effect of including these fibers in the diet to prevent some nowadays very frequent diseases. PMID- 9858131 TI - Aquaporins. AB - Aquaporins (AQP) are members of the major intrinsic protein superfamily of integral membrane proteins, which function as specialized water channels to facilitate the passage of water through the cell membrane in animals, plants and bacterias. Ten AQP homologues named from 0 to 9 have been clones so far in mammals. They are widely distributed and more than one AQP could be present in the same cell. Most of the AQPs are only permeable to water and impermeable to small organic and inorganic molecules, except for AQP 3, 7 and 9 which are also permeable to urea and glycerol. From the hydrophobicity profile all AQPs seem to have six transmembrane domains with five connecting loops and with the amino and carboxyl termini in the cytoplasm. They are synthesized as monomers, but there is evidence suggesting that AQPs are formed in the membrane as tetrameric units, each of which has four water pores. The primary amino acid sequence contains putative phosphorylation sites for protein kinasses A and/or C or casein kinase II, and the expression and membrane protein abundance of some AQPs are known to be under hormonal regulation. The human genes for several AQPs have been cloned and an increasing number of disturbances associated to abnormal functioning of these proteins had been identified. PMID- 9858132 TI - New insights into the immunoultrastructural organization of cutaneous basement membrane zone molecules. AB - The epidermal basement membrane zone (BMZ) is composed of various molecules, each of which plays an important role in dermo-epidermal adhesion. Genetic abnormality of certain BMZ molecules leads to an inherited group of skin diseases collectively referred to as epidermolysis bullosa, whose hallmark is skin fragility of varying degrees. Furthermore, development of autoantibodies to certain BMZ molecules leads to the onset of a number of acquired autoimmune blistering diseases in which dermo epidermal separation occurs, including bullous pemphigoid and epidermolysis bullosa acquisita. The ultrastructural location of each BMZ molecule has been studied using a range of immunoelectron microscopy (immuno-EM) techniques. Recent technical advances in immuno-EM and in molecular engineering for production of epitope-specific antibodies have enabled a more correct and precise elucidation of the native ultrastructural molecular organization of the respective molecules and their relationship to each other. These recent studies have also revealed several misinterpretations in the previously established model of the immunoultrastructural organization of BMZ molecules. In response to these findings, this review focuses on three major BMZ related molecules, type VII collagen, BPAG2 and laminin 5, for which recent immuno-EM studies have produced a revision in the accepted dogma on their ultrastructural distribution at the BMZ. PMID- 9858133 TI - Cytokines and cutaneous T-cell lymphomas. AB - Cytokines are considered to be of major importance for the pathogenesis of cutaneous T-cell lymphomas (CTCL). Their impact may result from autocrine, paracrine or endocrine effects. Several investigations demonstrated the overexpression of different cytokines in different CTCL entities. Interestingly, stage-dependent shifts in the cytokine pattern have been observed in mycosis fungoides (MF). There is evidence that the abnormal cytokine expression in CTCL might be responsible for tumor progression, resulting from an enhanced proliferation of the malignant cells and/or the depression of the anti-tumor immune response. Moreover, cytokine loops might explain phenomena like the epidermotropism of malignant cells or eosinophilia and increased plasma levels of IgE, which are present in advanced stages of CTCL. Analysis of the cytokine pattern in CTCL might give the basis for direct therapeutic intervention into the cytokine network as a new therapeutic approach. In this review, the current knowledge regarding cytokines in CTCL is summarized. PMID- 9858134 TI - A novel in-situ-zymography technique localizes gelatinolytic activity in human skin to mast cells. AB - Matrix-metallo-proteinases play a key role in cutaneous tissue remodeling and wound healing, and have been implicated as the rate-limiting factor in cutaneous tumor invasion and metastasis. We here describe a novel in-situ-zymographic method, which allows to directly localize sites of gelatinolytic activity in human skin. Gelatinolysis was detected through protein-hydrolysis in a 200 microm thick polyacrylamide gel underlying tissue sections. The lysis was substrate dependent, demonstrated time- and temperature-dependent kinetics, and was inhibited by both EDTA and 1,10-phenanthroline. Normal and diseased skin sections demonstrated multiple focal points of gelatinolysis which co-localized with individual cells. Histochemically, these were shown to represent most likely mast cells (via AS-d-chloroacetate esterase staining and metachromasia). However, immunohistochemical staining for gelatinases A and B showed no immunoreactivity patterns that corresponded to the identified foci of gelatinolysis. The reported in-situ-zymographic technique offers a decisive advantage over immunohistochemistry, since it detects only the activated and catabolically relevant proteases, and provides further evidence for a role of mast cells in extracellular matrix remodeling. PMID- 9858135 TI - An increased ratio of interleukin-1 receptor antagonist to interleukin-1alpha in inflammatory skin diseases. AB - IL-1 receptor antagonist (IL-1ra) is a cytokine that competitively binds the IL-1 receptor to antagonize IL-1 activity without any agonist function. Previous experiments indicated that the ratio of IL-1ra to IL-1alpha in the normal stratum corneum (SC) was much higher in the sun-exposed face than in the sun-protected area, upper arms. It was also reported by another laboratory that IL-1ra is increased in the lesional skin of psoriatic patients. This study was designed to measure the contents of IL-1alpha and IL-1ra in non-lesional and pathological SC obtained from inflammatory skin diseases including psoriasis and non-psoriatic dermatoses such as atopic dermatitis. The SC materials were obtained with a non invasive tape-stripping method. Their soluble fractions were prepared and assayed for IL-1alpha and IL-1ra by enzyme-linked immunosorbent assays. As a result we confirmed the previous findings that the ratio of IL-1ra to IL-1alpha in the normal SC was much higher in the face than in the sun-protected sites, the trunk as well as extremities. Next, we found that IL-1alpha contents were significantly reduced in the SC samples obtained from inflammatory skin regardless of whether their IL-1ra contents increased or unchanged. Moreover, we noted that an increased ratio of IL-1ra to IL-1alpha in the SC was not specific to psoriasis, but was also found in other inflammatory skin diseases including atopic dermatitis. This ratio was found to become lower after successful treatment of these skin lesions with topical glucocorticoids. We conclude from these observations that the increased ratio of IL-1ra to IL-1alpha in the SC is a non specific phenomenon that can occur in any inflammatory skin diseases regardless of the inflammatory pattern, probably reflecting a skin regulation process against various kinds of inflammation. PMID- 9858136 TI - In vivo human melanoma cytokine production: inverse correlation of GM-CSF production with tumor depth. AB - Melanomas produce multiple cytokines which may influence their growth in vivo. Experimental evidence suggests that granulocyte macrophage-colony stimulating factor (GM-CSF) can induce a potent anti-melanoma response. whereas interleukin-8 (IL-8) may act as a growth factor in human melanoma. Little is currently known regarding the production of these cytokines by human melanoma in vivo. In this study we tested the hypothesis that endogenous production of GM-CSF and IL-8 can be correlated with the depth of human malignant melanoma surgical specimens. We examined 45 melanocytic human tissue samples consisting of 27 primary cutaneous melanomas, 9 metastatic melanomas, and 9 dysplastic nevi for in vivo GM-CSF and IL-8 production using immunohistochemistry. The majority of thin melanomas (< or = 0.76 mm) stained highly positive for GM-CSF with little or no staining for IL-8 whereas the medium (>0.76- < or = 4.0 mm) and thick (>4.0 mm) melanoma specimens showed little or no staining for GM-CSF and significant amounts of IL-8 staining. Metastatic melanoma as well as dysplastic nevi specimens had little or no GM-CSF and IL-8 staining. These results support the hypothesis that endogenous melanoma cytokines such as GM-CSF and IL-8 with opposing effects on tumor progression play an important role in melanoma growth and regulation. PMID- 9858137 TI - Induction of apoptosis by synthetic ceramide analogues in the human keratinocyte cell line HaCaT. AB - In contrast to extracellular, long chain ceramides which comprise a structural component of the epidermal water barrier, intracellular ceramides originating from sphingomyelin hydrolysis have been shown to inhibit proliferation and to induce apoptosis in different cell populations. To further elucidate the possible role of intracellular ceramides in human epidermis, two new cell-permeable ceramide analogues, N-thioacetylsphingosine (C2-Cer=S) and 4-dodecanoylamino decan-5-ol (FS-5), were synthesized and tested for their ability to suppress cell growth and to induce apoptosis in immortalized human keratinocytes. It was shown that the well-investigated ceramide analogue N-acetylsphingosine (C2-Cer=O), as well as the new compound C2-Cer=S inhibited proliferation of HaCaT cells with half-inhibitory concentrations (IC50) of 20 microg/ml and 10 microg/ml, respectively, whereas FS-5 has been potent with an IC50>40 microg/ml. Overall, all three ceramide analogues induced apoptosis in HaCaT cells as assessed by DNA fragmentation using ELISA technique and in situ nick end labelling, thereby confirming the importance of ceramide signalling in keratinocytes. PMID- 9858138 TI - Lack of intrinsic polarity in the ligand-binding ability of keratinocyte beta1 integrins. AB - Within the basal layer of the epidermis the beta1 integrins have a pericellular distribution. Two monoclonal antibodies, 15/7 and 12G10, that detect a conformation of the beta1 integrin subunit that is induced following cation or ligand occupancy selectively recognized beta1 integrins at the basement membrane zone in vivo and in focal adhesions of cultured keratinocytes; they did not recognize integrins on the apical and upper lateral membranes of basal keratinocytes nor integrins on the suprabasal keratinocytes of hyperproliferative epidermis. Inhibition of intercellular adhesion did not induce the 15/7 epitope on the lateral and apical membrane domains. The surface distribution of the epitopes was consistent with the antibodies acting as reporters of ligand binding; in addition, the 15/7 epitope was exposed on unglycosylated, immature beta1 integrins. Although the apical membrane of basal keratinocytes is not normally in contact with extracellular matrix proteins, we found that it was capable of binding fibronectin-coated beads and that the 15/7 epitope was exposed on plasma membrane in contact with the beads. When a chimeric molecule consisting of the extracellular domain of CD8 and the cytoplasmic domain of the beta1 integrin subunit, used to mimic a constitutively active beta1 heterodimer, was introduced into keratinocytes it localized to the basal, lateral and apical membrane domains. We conclude that although the conformation of the keratinocyte beta1 integrins differs between the basal and the lateral/apical membrane domains there is no intrinsic polarity in the ligand binding potential of the receptors. PMID- 9858139 TI - Effects of UV irradiation with one minimal erythema dose on human afferent skin lymph in vivo. AB - Ultraviolet (UV) irradiation of the skin induces complex local and systemic immunomodulatory reactions. The biological effects of UV irradiation on human skin derived afferent lymph however are unknown. The aim of this study was to examine the effects of a single combined UV-A and UV-B irradiation with 1 minimal erythema dose (MED) on human skin derived lymph in vivo. After cannulation of a superficial lymph vessel on the lower leg, lymph flow and cell output per hour were determined before and for 6 days after UV irradiation of the lymph draining skin area in 5 volunteers. Furthermore, expression of CD1a, CD4, CD8, CD28, CD54, CD80, CD86 and HLA-DR on migrating lymph cells and cytokine levels (IL-1alpha, IL 1beta, IL-2, IL-6, IL-8, IL-10, IL-13, TNF-alpha and IFN-gamma) in the afferent lymph were analyzed by cytofluorometry and ELISA. After UV irradiation a small initial enhancement in the daily lymph flow per hour was noticed in correlation with the slight erythematous skin reaction. Following resolution of the skin reaction, a delayed increase in cell output in correlation with an additional peak in the lymph flow was found between the 4th and 6th day after UV irradiation. However, no changes in the expression of CD1a, CD4, CD8, CD28, CD54, CD80, CD86 and HLA-DR on migrating lymph cells were detectable. Interestingly, in parallel to the increased lymph flow and cell output, only elevated IL-8 protein levels were reproducibly detected in the afferent lymph after UV irradiation. Furthermore, using immunohistochemistry positive staining for IL-8 was found on migrating mononuclear lymph cells. In conclusion, our data demonstrate that a single UV irradiation of the skin with 1 minimal erythema dose leads to a delayed enhancement of lymph flow, number of migrating lymph cells and cytokine levels of IL-8. Moreover, we provide evidence that migrating lymph cells, besides resident epidermal and dermal cells, may contribute to the detected levels of IL-8 in the afferent lymph. PMID- 9858140 TI - Keratinocytes and dermal factors activate CRABP-I in melanocytes. AB - Recognition that cellular retinoic acid binding protein (CRABP)-I and CRABP-II are found in different cell types has provided additional support for the presumably divergent roles of these two proteins in mediating retinoic acid (RA) effects in human skin. CRABP-II is expressed in fibroblasts and keratinocytes, and CRABP-I in as yet unidentified cells, possibly epidermal melanocytes. Recently, we demonstrated that each of these RA-binding proteins in human skin possesses two classes of binding sites, possibly related to the state of phosphorylation of the proteins. We now characterize the cutaneous origin of CRABP-I further using an anion-exchange HPLC assay that allows effective separation of the two proteins in human skin, and a fluorescent in situ hybridization technique. We report that CRABP-I is expressed in isolated melanocytes at the mRNA level, although under these circumstances the protein has minimal RA-binding activity, and that keratinocytic and dermal influences are required for CRABP-I activity in melanocytes. This melanocyte origin for CRABP-I and the improvement by RA of the irregular hyperpigmentation associated with photoaging led us to examine the effects of RA using various cellular associations, from conventional pure cultures of melanocytes grown on plastic dishes to a pigmented skin equivalent consisting of melanocytes and keratinocytes grown on a dermal equivalent. We established that the inhibitory effects of RA on melanogenesis do not result from a direct effect on melanocytes alone but also involve keratinocytes and dermal influence. These data expand our understanding of cell-to-cell signaling in cutaneous pigmentation, and strongly suggest a role for CRABP-I in mediating RA effects on melanogenesis. PMID- 9858141 TI - Similarities of cutaneous and breast pathology in Cowden's Syndrome. AB - Cowden's Syndrome (CS), or multiple hamartoma syndrome, is an autosomal dominant disorder characterized by mucocutaneous lesions, multiple benign tumors of internal organs and an increased risk of breast cancer. Here, we describe and illustrate in detail the benign breast pathology of 59 breast cases from 19 women with CS. Fibrosis is a significant characteristic of the breasts of patients with CS. Fibroadenomas appear to hyalinize at an early age and are frequently complex. The specimens demonstrate a spectrum of dense hyalinization of both the lobule and the stroma, and hyaline nodules appear to be the most characteristic lesion. This hyalinization process shares striking similarities with keloids, as well as the sclerotic nodules seen in the skin of CS individuals. Ductal carcinoma in CS was common, and it appeared to be associated with stromal hyalinization. Other frequently found benign features are adenosis and cysts. Of interest, the features of the benign breast disease in CS show certain similarities with senescent lobules, fibrous mastopathy of diabetes mellitus, and mammary hamartomas. These observations provide a framework for pathologists to identify this underrecognized syndrome. PMID- 9858142 TI - Retinoid regulation of heparin-binding EGF-like growth factor gene expression in human keratinocytes and skin. AB - Retinoic acid (RA) has profound effects on epidermal homeostasis; however, the molecular mechanisms by which retinoids regulate keratinocyte cell proliferation and differentiation are not well understood. Here we report that mRNA expression of heparin-binding EGF-like growth factor (HB-EGF), a member of the EGF family of growth factors, is induced by RA in human keratinocytes and skin, and is overexpressed in the context of epidermal hyperplasia in vivo. Treatment of normal adult human keratinocytes with micromolar concentrations of RA significantly induced the expression of HB-EGF. The response was efficiently blocked by specific inhibitors of ErbB tyrosine kinase activity, MAP kinase kinase (MEK), or p38 stress-activated protein kinase. RA also enhanced the induction of HB-EGF mRNA in human skin organ culture, an ex vivo model system displaying many similarities to wound healing in vivo. HB-EGF transcripts were markedly increased in human skin by topical treatment with RA under conditions known to provoke epidermal hyperplasia. HB-EGF transcripts were also markedly overexpressed in the hyperplastic epidermis of psoriatic lesions, relative to normal skin. These results support the hypothesis that the effects of RA on epidermal hyperplasia are mediated at least in part by HB-EGF, and suggest that signal transduction mechanisms other than or in addition to nuclear RA receptors contribute to this effect. PMID- 9858143 TI - The human immunodeficiency virus (HIV)-type 1 coreceptor CXCR-4 (fusin) is preferentially expressed on the more immature CD34+ hematopoietic stem cells. AB - In synergy with the CD4 antigen, the chemokine receptor CXCR-4 functions as a coreceptor for T-cell-tropic HIV-1 strains. Using two- and three-color immunofluorescence analysis, we examined the expression of CXCR-4 on CD34+ cells in 21 samples obtained from leukapheresis (LP) products of cancer patients who underwent G-CSF-supported cytotoxic chemotherapy. In addition, eight samples from bone marrow (BM) were obtained. CXCR-4 was expressed on the surface of CD34+ cells from samples of both hematopoietic sources. The mean proportion of CD34+/CXCR-4+ cells from LP products was 1.7-fold greater in comparison with those from bone marrow (65.9+/-4.1% vs. 37.5+/-8.6% [+/- SEM], p < 0.05). For an intraindividual comparison, LP products and bone marrow from six patients were obtained on the same day, confirming the significantly greater proportion of CD34+ cells coexpressing CXCR-4 cells in LP products. In order to examine whether the CXCR-4 expression was related to the stage of maturation and differentiation of CD34+ cells, six samples from LP products and four samples from bone marrow were assessed using three-color immunofluorescence analysis. We found that the subset of CD34+/CD38low and CD34+/HLA-DRlow cells representing a population of more immature progenitor cells were brightly positive for CXCR-4, while there was a decrease in the level of CXCR-4 expression in the population of CD34+/HLA DRbright and CD34+/CD38bright cells. Based on the assessment of ten LP products, we found that the mean proportion of CD34+ cells coexpressing CD4 and CXCR-4 was 6.2+/-2.3% [+/- SEM], suggesting that a small population of CD34+ cells are, in principle, susceptible for an infection with T-cell-tropic HIV-1 strains. In conclusion, our data suggest that CXCR-4 is present on the surface of hematopoietic progenitor cells--particularly more primitive CD34+ cells. CXCR-4 could play a role in the homing of CD34+ cells to stromal elements of the bone marrow via its natural ligand stromal-derived factor-1 (SDF-1). PMID- 9858144 TI - Regulation and possible function of axl expression in immature human mast cells. AB - The receptor tyrosine kinase Axl which expresses extracellular domains reminiscent of cell adhesion molecules, is involved in homotypic binding as well as in intracellular signaling of myeloid progenitor cells. In order to investigate factors which might influence differentiation pathways through changes of the adhesive properties of cells, we analyzed the expression of axl in immature basophil and mast cell lines and in cultured basophil and mast cell precursors. Axl expression was induced by interferon-alpha in the human leukemic mast cell line HMC-1 and in cultured mast cells derived from CD34+ peripheral blood cells. Axl induction was dose dependent, appeared within 1 h, and was independent of de novo protein synthesis. IFNalpha-treated HMC-1 cells expressing axl formed large cell aggregates within 40 h while untreated cells did not. HMC-1 cells also expressed gas6, the putative ligand of axl, which has been shown to induce axl-mediated homotypic binding. Gas6 expression was independent of interferon treatment in HMC-1 cells. The present results suggest that axl mediated changes of cellular adhesive properties in mast cells may be important in mast cell differentiation as well as in mast cell-associated inflammation. PMID- 9858145 TI - Differential expression of the RET gene in human acute myeloid leukemia. AB - The RET proto-oncogene product is a receptor tyrosine kinase representing the signal-transducing molecule of a multi-subunit membrane receptor complex for at least two different types of transforming growth factor (TGF)-beta-related neurotrophic factors. We have previously shown that RET gene expression in acute myeloid leukemia (AML) occurs more frequently in AMLs displaying either a monocytic (FAB M4/M5) or intermediate-mature myeloid phenotype (FAB M2/M3) than in leukemias reflecting an earlier stage of myeloid differentiation (FAB M0/M1). To further verify the association between RET expression and the relative maturation stage of AML cells, we have performed a quantitative estimation of relative abundances of RET transcripts among various FAB subtypes of AMLs. By analyzing 13 AML samples and normal hematopoietic cells through a competitive quantitative RT-PCR approach, we were able to show that the relative levels of RET-specific mRNAs continuously increase with blast cell maturation in human AML, i.e., the amounts of RET gene-specific transcripts differ among RET-expressing AMLs, being higher in the more differentiated FAB phenotypes. In addition, we provide evidence that the relative amounts of RET transcripts increase upon in vitro and in vivo differentiation of leukemic promyelocytes from FAB M3 AML patients, becoming overall comparable to those found in normal granulocytes. These results indicate that RET expression in human AMLs is maturation associated, probably mirroring the developmental regulation of this gene during differentiation of normal hematopoietic cells. PMID- 9858146 TI - High levels of thrombopoietin in sera of patients with essential thrombocythemia: cause or consequence of abnormal platelet production? AB - Thrombopoietin (TPO) is the most important regulator of megakaryocyte development and platelet production. Platelet production is thought to be regulated by a negative regulatory feed back loop. In an attempt to evaluate the role of TPO in the pathobiology of essential thrombocythemia (ET), we have examined levels of TPO and other cytokines with thrombopoietic activity (interleukin-6 and interleukin-11) in sera obtained from 25 patients with ET (ten treated, 15 untreated) and 117 healthy control subjects. TPO serum levels were assessed using a sandwich-antibody ELISA that utilizes a polyclonal rabbit antiserum for both capture and signal. The mean serum TPO level in 25 ET patients was significantly elevated (545+/-853 pg/ml) as compared with that in healthy controls (95.3+/-54.0 pg/ml,p<0.001). The difference in TPO serum levels between ten treated (781+/ 1229 pg/ ml) and 15 untreated ET patients (388+/-458 pg/ml) did not reach statistical significance (p = 0.09). We conclude that either consumption or production of TPO is altered in ET. Failure of appropriate feedback regulation and continued megakaryocyte stimulation by an elevated TPO may play an important role in the pathobiology of ET. PMID- 9858147 TI - Thrombopoietin serum concentration in patients with reactive and myeloproliferative thrombocytosis. AB - We wished to test whether thrombopoietin (TPO) is entirely regulated by receptor binding or if other factors may play a role in the mechanism of TPO regulation. Therefore, we analyzed the TPO serum levels in 43 patients with reactive (secondary) thrombocytosis and in 37 with myeloproliferative thrombocytosis. Thrombocytosis was defined as a platelet level greater than 440 x 10(9)/l. Forty two patients (98%) with reactive thrombocytosis had high concentrations of IL-6 correlating with elevated C-reactive protein levels. Twenty-three patients (53%) in this group had TPO serum concentrations of more than 300 pg/ml (normal: below 300 pg/ml). Only nine patients (24%) with myeloproliferative thrombocytosis had TPO serum levels above normal range, whereas 28 patients (76%) had normal levels of TPO. No correlation between the TPO serum levels and the concentrations of IL 6 or EPO was established. The other investigated thrombopoietic cytokines (IL-3, IL-11, GM-CSF) were unmeasurable; therefore, a correlation could not be assessed. We conclude that TPO concentrations are not strictly inversely related to platelet count. TPO serum levels are elevated especially in a considerable percentage of patients with reactive thrombocytosis, arguing for the existence of additional mechanisms of TPO regulation. PMID- 9858148 TI - Idarubicin and intermediate-dose cytarabine for myeloid blast crisis of chronic myelogenous leukemia--results of a phase-II trial. AB - Sixteen patients with Philadelphia chromosome-positive chronic myelogenous leukemia (CML) in myeloid blast crisis were treated with cytarabine (AraC) 600 mg/m2 two times daily for 5 days and idarubicin 12 mg/m2 for 3 days. Patients achieving a second chronic phase received interferon (IFN) alpha 2b 5 mio units/day daily and AraC 20 mg/day subcutaneously 14 days every month. Study end points were remission rate and survival. Four patients (25%) entered a second chronic phase and had a median survival of 31.1 weeks (range 16.1-111 weeks). Nine patients (56%) experienced blast crisis again and had a median survival of 12.9 weeks (range 5.1-59.3 weeks). Three patients (18.8%) died of septic complications during marrow aplasia. The median overall survival was 16.1 weeks (range 2.6-111 weeks) with no significant difference between responders and nonresponding patients. We conclude that AraC/idarubicin is as effective as other intensive regimens in inducing second chronic phase in patients with myeloid blast crisis of CML. Remission duration and survival are comparable to previous results. Further studies to improve survival are required. PMID- 9858149 TI - A case of mu heavy-chain disease associated with hyperglobulinemia, anemia, and a positive Coombs test. AB - We report here for the first time a patient with mu heavy-chain disease (HCD), hyperimmunoglobulinemia, and a positive direct antiglobulin test (DAT, Coombs test). The heavy-chain diseases involve the proliferation of lymphoplasma cells of B cell origin and are characterized by the production of incomplete heavy chains devoid of light chains. The association of mu heavy-chain disease with either hyperglobulinemia or a positive DAT has not been reported in the literature to date. In this patient, immunofixation of serum proteins with monospecific antisera to alpha-, gamma-, mu,- or delta-chains and to kappa- and lambda-chains revealed a precipitation band with antibody to IgM, but not with kappa and lambda light-chain antibodies, indicating mu heavy-chain disease. Hyperglobulinemia was present, which is very uncommon for HCD. A DAT of the patient's red blood cells (RBC) was found to be strongly positive for anti-IgG but negative for anti-IgM, -IgA, -C3c, and -C3d. However, when the eluate from the patient's red blood cells was investigated with nephelometry, it was found to contain antigens reactive with anti-y as well with anti-mu-antiserum. When a DAT was performed with a randomly chosen test cell incubated with the eluate, the antibody-containing eluate was shown to react with anti-IgG as well as with anti IgM-antiserum. In summary, the eluate from the patient's RBCs contained IgG and an immunoglobulin structure reactive with anti-IgM in an RBC agglutination assay as well as with anti-mu antiserum in a nephelometric investigation. Whether this IgM on the patient's erythrocytes is penta- or oligomeric, complete IgM, or the heavy chain cannot be concluded from these observations. PMID- 9858150 TI - L-tryptophan-related eosinophilia-myalgia syndrome possibly associated with a chronic B-lymphocytic leukemia. AB - A female patient presenting with B-CLL and coincident eosinophilia-myalgia syndrome (EMS) after ingestion of L-tryptophan is described. The manifestations of EMS disappeared completely during treatment with cyclophosphamide/prednisone. and there was an intermittent clinical remission of CLL with absence of the monoclonal B-cell population. A few years later, the B-CLL relapsed, but without sign and symptoms of EMS. Whereas other eosinophilic syndromes such as eosinophilic fasciitis, panniculitis, or cellulitis Wells have been found to occur in relation to malignant underlying diseases, only a single patient with malignant fibrous histiocytoma following EMS has been described. There are no reports about an increased occurrence of B-CLL or other non-Hodgkin's lymphomas combined with or following EMS or related to L-tryptophan itself. The variant types of eosinophilic syndromes occurring due to malignant disorders, the differentiation from EMS, and the possible association between B-CLL and L tryptophan-related EMS are discussed. PMID- 9858151 TI - Transient tetraparesis after intrathecal and high-dose systemic methotrexate. AB - Aggressive polychemotherapy, intrathecal cytostatic prophylaxis and cranial irradiation have contributed to the remarkable improvement in the prognosis of acute lymphoblastic leukemia (ALL) and subtypes of high-grade non-Hodgkin's lymphoma (NHL) and the reduction of central nervous system (CNS) relapses. Early and late neurologic changes have been observed after different CNS-directed therapies. We report on the rare event of an acute tetraparesis after methotrexate (MTX) without other CNS-directed therapy. A young female with a diffuse large B-cell lymphoma developed signs of meningeal irritation a few hours after intrathecal prophylaxis with MTX, cytosine-arabinoside and dexamethasone. She recovered quickly. Ten days after her last course of systemic chemotherapy including high dose MTX she was admitted with a tetraparesis and motoric aphasia. A computer assisted tomography (CT) scan was normal. On magnetic resonance imaging (MRI) hyperintense white matter lesions were visible in the periventricular white matter. Initially, the radiologic signs were progressive while the patient's clinical condition improved. MRI controls after complete neurologic normalization revealed delayed partial regression of the white matter abnormalities. The patient has now been free of neurologic symptoms for 16 months. This case report demonstrates acute and subacute neurotoxic effects of MTX in the same patient and illustrates that radiologic CNS changes can persist irrespective of the disappearance of clinical symptoms. PMID- 9858152 TI - Interleukin-6 may play an important role in thrombopoiesis: a case of leukemic transformation from myelodysplastic syndrome. PMID- 9858153 TI - Supernumerary centrosomes and cancer: Boveri's hypothesis resurrected. PMID- 9858154 TI - SF-assemblin, striated fibers, and segmented coiled coil proteins. AB - The flagellar basal apparatus of many flagellate green algae contains noncontractile striated microtubule-associated fibers (SMAFs). The SMAFs consist of 2-nm protofilaments and are predominantly built from striated fiber (SF) assemblin, an acidic 33-kDa protein. In this review we summarize the present knowledge concerning the biochemical properties of SF-assemblin and the molecular architecture of the SMAFs, provide evidence for homologous proteins and similar filament systems in other eukaryotes, and, finally, discuss possible biological functions of SF-assemblin. PMID- 9858155 TI - Flagellar coordination in Chlamydomonas cells held on micropipettes. AB - The two flagella of Chlamydomonas are known to beat synchronously: During breaststroke beating they are generally coordinated in a bilateral way while in shock responses during undulatory beating coordination is mostly parallel [Ruffer and Nultsch, 1995: Botanica Acta 108:169-276]. Analysis of a great number of shock responses revealed that in undulatory beats also periods of bilateral coordination are found and that the coordination type may change several times during a shock response, without concomitant changes of the beat envelope and the beat period. In normal wt cells no coordination changes are found during breaststroke beating, but only short temporary asynchronies: During 2 or 3 normal beats of the cis flagellum, the trans flagellum performs 3 or 4 flat beats with a reduced beat envelope and a smaller beat period, resulting in one additional trans beat. Long periods with flat beats of the same shape and beat period are found in both flagella of the non-phototactic mutant ptx1 and in defective wt 622E cells. During these periods, the coordination is parallel, the two flagella beat alternately. A correlation between normal asynchronous trans beats and the parallel-coordinated beats in the presumably cis defective cells and also the undulatory beats is discussed. In the cis defective cells, a perpetual spontaneous change between parallel beats with small beat periods (higher beat frequency) and bilateral beats with greater beat periods (lower beat frequency) are observed and render questionable the existence of two different intrinsic beat frequencies of the two flagella cis and trans. Asynchronies occur spontaneously but may also be induced by light changes, either step-up or step down, but not by both stimuli in turn as breaststroke flagellar photoresponses (BFPRs). Asynchronies are not involved in phototaxis. They are independent of the BFPRs, which are supposed to be the basis of phototaxis. Both types of coordination must be assumed to be regulated internally, involving calcium sensitive basal-body associated fibrous structures. PMID- 9858156 TI - Human brush border myosin-I and myosin-Ic expression in human intestine and Caco 2BBe cells. AB - The human intestinal cell line, Caco-2BBe, has been established as an excellent model system for analysis of the enterocyte cytoskeleton including that of the actin rich apical brush border. To facilitate its use for functional analysis of a major component of the brush border, brush border myosin-I, human cDNAs encoding the heavy chain of this class I myosin were isolated and sequenced. The identity of this myosin as human brush border myosin-I was verified based on similarity with other vertebrate sequences, as well as its expression profile at both the RNA and protein levels. Localization of the protein in human intestine along the crypt-villus axis closely resembles that previously determined for brush border myosin-I in chicken, and is quite distinct from that of myosin-Ic, another myosin-I expressed in human intestine and Caco-2BBe cells. In immature cells of the crypt, brush border myosin-I staining is low, and there is significant cytosolic and basolateral localization, while villus cells stain much more intensely, and the protein is primarily localized to the brush border. Localization of myosin-Ic is essentially the inverse of brush border myosin-I in that crypt cells exhibit higher levels of staining, while villus cells have very low levels of myosin-Ic. The expression of both myosins-I was also examined during cell-contact induced differentiation of Caco-2BBe cells where expression and changes in localization closely resemble those that accompany differentiation of enterocyte in vivo. PMID- 9858157 TI - Relationship between microtubule dynamics and lamellipodium formation revealed by direct imaging of microtubules in cells treated with nocodazole or taxol. AB - Microtubules (MTs) contribute to the directional locomotion of many cell types through an unknown mechanism. Previously, we showed that low concentrations (<200 nM) of nocodazole or taxol reduced the rate of locomotion of NRK fibroblasts over 60% without altering MT polymer level [Liao et al., 1995: J. Cell Sci. 108:3473 3483]. In this paper, we directly measured the dynamics of MTs in migrating NRK cells injected with rhodamine tubulin and treated with low concentrations of nocodazole or taxol. Both drug treatments caused statistically significant reductions (approx. twofold) in growth and shortening rates and less dramatic effects on rescue and catastrophe transition frequencies. The percent time MTs were inactive (i.e., paused) increased greater than twofold in nocodazole- and taxol-treated cells, while the percent time growing was substantially reduced. Three parameters of MT dynamics were linearly related to the rates of locomotion determined previously: rate of shortening, percent time pausing and percent time growing. The number of MTs that came within 1 microm of the leading edge was reduced in drug-treated cells, suggesting that reduced MT dynamics may affect actin arrays necessary for cell locomotion. We examined two such structures, lamellipodium and adhesion plaques, and found that lamellipodia area was coordinately reduced with MT dynamics. No effect was detected on adhesion plaque density or distribution. In time-lapse recordings, MTs did not penetrate into the lamellipodium of untreated cells, suggesting that MTs affect lamellipodia either through their interaction with factors at the base of the lamellipodium or by releasing factors that diffuse into the lamellipodia. In support of the latter hypothesis, when all MTs were rapidly depolymerized by 20 microM nocodazole, we detected the rapid formation of exaggerated protrusions from the leading edge of the cell. Our results show for the first time a linear relationship between MT dynamics and the formation of the lamellipodium and support the idea that MT dynamics may contribute to cell locomotion by regulating the size of the lamellipodium, perhaps through diffusable factors. PMID- 9858158 TI - Expression of glycylated tubulin during the differentiation of spermatozoa in mammals. AB - Using quantitative immunogold analyses of tubulin isoforms we previously demonstrated a unique differential expression of glutamylated tubulin in the flagellum of mouse and man spermatozoa [Fouquet et al., 1997: Tissue Cell 29:573 583]. We have performed similar analyses for glycylated tubulin using two monoclonal antibodies, TAP 952 and AXO 49, directed to mono- and polyglycylated tubulin respectively. Glycylated tubulin was not found in centrioles and cytoplasmic microtubules (manchette) of germ cells. In mouse and man, axonemal tubulin was first monoglycylated and uniformly distributed in all doublets at all levels of the flagellum in elongating spermatids. In human mature spermatozoa axonemal microtubules were enriched in monoglycylated tubulin from the base to the tip of the flagellum. In mouse sperm flagellum a similar gradient of monoglycylated tubulin was also observed in addition to an opposite gradient of polyglycylated tubulin. In both species, monoglycylated tubulin labeling predominated in doublets 3-8 whereas glutamylated tubulin labeling [Fouquet et al., 1997] predominated in doublets 1-5-6. These differential labelings were suppressed after motility inhibition of mouse spermatozoa by sodium azide treatment and in non-motile human spermatozoa lacking dynein arms. The unique distribution of these tubulin isoforms and the known inhibition of motility induced by their specific antibodies are consistent with a complementary role of tubulin glycylation and glutamylation in the regulation of flagellar beating in mammalian spermatozoa. PMID- 9858159 TI - Isolation and characterization of novel Chlamydomonas mutants that display phototaxis but not photophobic response. AB - The unicellular green alga Chlamydomonas displays two distinct kinds of behavioral response to light: phototaxis, in which cells swim toward or away from the light source under constant illumination; and photophobic responses (also called stop responses or photoshock responses), in which cells transiently convert their flagellar waveform and swim backward upon sudden increase in light intensity. It has been suggested that the two responses partly share a common signal transduction pathway, but exactly how the different responses are produced has not been established. In this study, to help understand the molecular and cellular mechanisms that bring about the photophobic response, we isolated novel mutants (ppr1, ppr2, ppr3, and ppr4) that do not show the photophobic response. Importantly, these mutants retain the ability to display phototaxis, with almost the same sensitivities as in the wild type cell. Demembranated and reactivated flagellar axonemes of the ppr mutants were found to convert the bending patterns depending on the Ca2+ concentration, indicating that the axonemal mechanism for waveform conversion required for the photophobic response was unaffected by the mutations. In addition, measurements of electric currents in cell suspensions showed that these mutants generate normal photoreceptor currents (PRC) upon photostimulation, suggesting that they retain the normal activity of photoreception and the ionic channels that produce PRCs. However, the all-or-none flagellar current (FC), a Ca2+ current generated by PRC-induced depolarization of flagellar membrane, was absent or seriously impaired in the mutants. These findings clearly indicate that the all-or-none FC is necessary for the photophobic response but not for phototaxis. The isolation of the four genetically independent ppr mutants suggests that the generation of the FC is based on multiple components that are not used in the mechanism for phototaxis, and implies that the Chlamydomonas flagellar membrane possesses a voltage dependent Ca2+-channel specifically used for generation of photophobic responses. PMID- 9858160 TI - Dermal autografts for fascial repair after TRAM flap harvest. AB - To find an alternative to synthetic mesh closure of abdominal fascial defects after transverse rectus abdominis musculocutaneous (TRAM) flap harvest, dermal autografts were removed from tissue to be discarded and used for fascial closure. Dermal grafts have been used for herniorrhaphy and fascial repair after TRAM harvest previously, but have never been systematically studied. The dermal autograft technique was used in 24 patients to repair or reinforce anterior rectus sheath or external oblique fascia after TRAM harvest for breast reconstruction. During the same period, 25 other patients underwent TRAM breast reconstruction with abdominal wall closure by other methods. All patients were followed by serial physical examinations given by the operating surgeon. Average follow-up in the dermal autograft group was 12.6 versus 12.0 months in the second group. In the dermal autograft group, two patients complained of bulging of the anterior abdominal wall; one developed a true hernia, away from the location of the dermal autograft. In the second group, two patients experienced bulging. Wounds and infectious complications were similar in both groups. Dermal autografts are a useful alternative to mesh repair or direct closure of fascial defects after TRAM flap harvest. PMID- 9858161 TI - Tissue-engineered nipple reconstruction. AB - We describe a simple, effective approach to the creation of autologous tissue engineered cartilage in the shape of a human nipple by injecting a reverse thermosensitive polymer seeded with autologous chondrocytes in an immunocompetent porcine animal model. A biodegradable, biocompatible copolymer of polyethylene oxide and polypropylene oxide (Pluronic F-127), which exists as a liquid below 4 degrees C and polymerizes to a thick gel when it is exposed to physiologic temperatures (body temperatures), was used as a vehicle for chondrocyte delivery and as a scaffold to guide growth. Autologous chondrocytes isolated from porcine auricular elastic cartilage and suspended in 30% (weight/volume) Pluronic F-127 were injected on the ventral surface of the pigs from which the cells had been isolated. A circumferential subdermal suture was used to support the contour of the implant and assist in its projection in the form of a human nipple. After 3 weeks, the skin over and surrounding the implant was tattooed to create the appearance of a human nipple-areolar complex. As controls, an equal number of injections were made using either cells alone (not suspended in hydrogel), or hydrogel alone. After 10 weeks, all specimens were excised and examined both grossly and histologically. Before harvesting, visual inspection of the tattooed chondrocyte-Pluronic F-127 hydrogel implant sites revealed that they closely resembled a human female nipple-areolar complex. Nodules were similar in size, shape, and texture to a human nipple at each injection site. Glistening opalescent tissue was surgically isolated from each implant site. Hematoxylin and eosin, safranine o, trichrome blue, and Verhoeff's stains of the experimental implants showed nodules with the characteristic histologic signs of elastic cartilage. Control injections of copolymer hydrogel alone exhibited no evidence of cartilage formation. Control injections of chondrocytes alone showed evidence of dissociated microscopic nodules of elastic cartilage. PMID- 9858162 TI - Absence of longitudinal changes in rheumatologic parameters after silicone breast implantation: a prospective 13-year study. AB - There have been numerous questions regarding the association of polysiloxane with connective tissue disease and alteration of host immune response. C-reactive protein, rheumatoid factor, and anti-streptolysin-O titers were measured in 218 patients. These studies are routinely used in the diagnosis of autoimmune disease and mixed connective tissue disease. This prospective study has been in progress since 1985. The first patients were seen in July of 1985, and those individuals willing to participate were followed from 1985 to 1998. The implants included saline-filled elastomer shells and polysiloxane gel-filled elastomer shells. These groups were examined separately and in combination for changes between preoperative and postoperative states. In each instance, there was no statistical increase or decrease. Each patient underwent a physical examination and completed a questionnaire focusing on signs and symptoms of autoimmune and connective tissue diseases. The laboratory data and subjective clinical results demonstrated no significant differences between a nonimplanted group versus the saline group alone, the gel group alone, or the combined groups. The data failed to suggest any causal relationship between implants and autoimmune or connective tissue diseases over the study period of 13 years (since 1985). PMID- 9858163 TI - An analysis of silicone gel-filled breast implants: diagnosis and failure rates. AB - This retrospective review analyzed 180 women who underwent explantation of 357 silicone gel-filled breast implants from September of 1991 to January of 1995. Implant status, including shell integrity and implantation times, was determined at the time of explantation. The age range of the patients was 25 to 75 years, with a mean age of 47 years. The age of the implants ranged from 0.5 to 24 years, with a mean age of 10.5 years. Of the 292 implants with known and documented integrity status, there were 102 intact, 76 unruptured with signs of leakage, and 114 ruptured. The frequency of implant rupture significantly increased with implant age. The average age of rupture was 13.4 years. The average age of signs of leakage was 10.1 years. There were no significant differences in failure rates among the implant types of four manufacturers. Analysis of both mammography and magnetic resonance imaging (MRI) as diagnostic modalities for differentiating intact implants, implant leakage, and implant rupture was performed. Standard mammography was less reliable in diagnosing implant leakage or rupture (sensitivity, 55 percent; specificity, 69 percent) than MRI (sensitivity, 72 percent; specificity, 82 percent). In conclusion, implant rupture occurred at a significantly increasing rate with implant age (10 to 15 years). These findings were independent of implant type or manufacturer. Mammography alone is a below average diagnostic tool for detecting leakage or rupture, whereas MRI is a more accurate modality. PMID- 9858164 TI - The Le Fort I-palatal split approach for skull base tumors: efficacy, complications, and outcome. AB - The Le Fort I, split-palate approach provides intraoral surgical access to a region of the midline skull base ranging from the upper clivus to the second cervical vertebra. Although this approach provides perhaps the largest exposure of all the intraoral techniques, there is little concerning it in the literature. Furthermore, there are no detailed descriptions of case histories, complications, and outcome. The purpose of this study was to evaluate this procedure's effectiveness and identify associated complications as well as outcome. Seven cases of patients who underwent eight skull base surgeries using the Le Fort I, split-palate approach were evaluated retrospectively. Particular attention was paid to postoperative occlusion, speech, mouth opening, infection, tumor recurrence, postoperative recovery period, and viability of maxillary bone and teeth. Follow-up ranged from 4 months to 7 years with a mean of 3.9 years. Pathologic diagnoses included three chordomas (two recurrent), one recurrent meningioma, one liposarcoma, one chondrosarcoma, and one inflammatory mass. One patient with chordoma underwent a second operation using the same approach. No deaths or major neurologic problems related to the procedure occurred. One patient who had known local metastases at the time of operation died several months after surgery. All other patients are still living. Duration of hospital stay ranged from 5 to 53 days with a mean of 25.4 days. Postoperative complications included one case of meningitis with an associated cerebrospinal fluid leak, three cases of malocclusion, one case of velopharyngeal insufficiency, and one extracranial soft-tissue infection. The case of meningitis was successfully treated by antibiotics. The malocclusions were corrected by conservative treatment. No problems with mouth opening or bone or tooth viability occurred. Tumor recurred in both cases in which malignancy was involved, whereas only one recurrence was noted among the benign cases. It is concluded that the Le Fort I-palatal split technique is a relatively safe and effective means for approaching midline skull base tumors. Several modifications to the surgical protocol and surgical technique are detailed herein. PMID- 9858165 TI - Reconstruction of congenital stenosis of external auditory canal with a postauricular chondrocutaneous flap. AB - We describe here a new surgical technique for treatment of congenital stenosis of the external auditory canal using a postauricular chondrocutaneous flap. Our technique prevents recurring stenosis by cartilage support and provides a sufficient extension of the canal. The chondrocutaneous flap is safely elevated, easily reaches the canal, and has ample blood supply. It is beneficial for its self-cleaning function that the remnant canal, which contains cerumen glands, is used as part of the reconstructed canal. Six patients with congenital stenosis of the external auditory canal were treated by our procedure. In these patients, three patients were treated by a postauricular approach and the others were treated by an endaural approach. All patients had very satisfactory results. PMID- 9858166 TI - 5-year series of constricted (lop and cup) ear corrections: development of the mastoid hitch as an adjunctive technique. AB - Despite the multitude of corrective procedures described, adequate surgical correction of the congenital constricted ear remains a challenge. The maintenance of the shape and elevation of the reconstructed upper neohelix poses a particular problem. In the present series, experiences with lop ear correction utilizing standard techniques and the use of the mastoid hitch as a useful adjunct to these procedures are described. A total of 19 ears were reconstructed. There were three type 1, eight type 2a, seven type 2b, and one type 3 deformities (Tanzer classification). A graded sequence of procedures was adopted. Mild deformities were corrected by cartilage scoring techniques; a V-Y advancement of the helical root was added for moderate deformities. Cartilage expansion by a banner flap was required for more severe deformities. A mastoid hitch, whereby the refashioned upper neohelix is sutured to the mastoid fascia, should be used as an adjunct to these procedures to maintain helical elevation and prevent recurrence. Severe type 3 deformities may require autologous auricular reconstruction. Mean follow up time was 1 year. There were six excellent, seven good, four fair, and two poor results. Two patients who had not had mastoid hitch procedures developed a recurrence of the lop deformity. Adequate surgical correction of constricted ear deformities requires a variety of surgical techniques. The mastoid hitch being used for constricted ear correction has not been described elsewhere. The mastoid hitch is a useful adjunctive procedure that may be used effectively in combination with other procedures. PMID- 9858167 TI - A method that preserves circulation during preparation of the pectoralis major myocutaneous flap in head and neck reconstruction. AB - The present article describes a method that preserves circulation during the preparation of the pectoralis major myocutaneous flap used in head and neck reconstruction. The major disadvantage of this flap is its poor circulation and consequent partial necrosis. To solve this problem, we analyzed the circulation and hemodynamics of the pectoralis major myocutaneous flap (the perforator of the anterior intercostal branch located about 1 to 2 cm medial to the areola in the fourth intercostal space is important), evaluated the safe donor sites in the chest wall for a skin island (the perforator is included on the skin island's central axis), improved the surgical procedure for elevating flaps (for preventing perforator injuries), and devised a means to transfer flaps, thereby increasing the range of the flaps (the transfer route is under the clavicle). Using this technique, head and neck reconstruction was performed on 62 patients. The diagnosis included oral cancer (21), oropharyngeal carcinoma (10), parotid carcinoma (10), hypopharyngeal carcinoma (9), and other head and neck malignant tumors (12). Of these, partial or marginal necrosis of the flap caused by circulatory problems was detected in three patients (5 percent). Using our method, the problems associated with inadequate circulation in the pectoralis major myocutaneous flap were greatly alleviated, thus reconfirming the usefulness of this flap in head and neck reconstruction. PMID- 9858168 TI - Free transverse colon transplantation for functional reconstruction of intra-oral lining: a clinical and histologic study. AB - A variety of free-tissue transplantations are available for oropharyngeal (lining) reconstruction. The most commonly used flap, the radial forearm flap, is relatively thin, adaptable, and reliable but does not provide lubrication. In this study, the potential of the transverse colon to provide a lubricating tissue for replacement of oral lining was investigated. Eleven patients requiring replacement of oral lining received transverse colon flap transplantation between November of 1993 and December of 1995. There was one complete flap loss and one partial loss. Flap size used for reconstruction averaged 40.4 cm2, and average vessel length and diameter were 7 cm and 2.0 mm, respectively. In a follow-up period from 15 to 48 months, all colon flaps proved to be durable with continuous lubrication function. Histologic assessment of the transplanted colon flaps at various postoperative times showed an adequate amount of mucus-secreting goblet cells even after irradiation. There was no donor-site morbidity. The main disadvantage seemed a tendency to transplant too much tissue, which resulted in formation of redundant pockets affecting food handling. Thus, 8 out of 10 patients with flap survival required one to three debulking procedures. Given an accurate estimation of the area of mucosa defect and adequate trimming of the colon flaps, the transverse colon flap offers a good alternative for reconstruction of the oral lining, especially when lubrication is desirable in cases with large defects and preoperative or postoperative irradiation, or in case a radial forearm flap is not available. PMID- 9858169 TI - Efficacy of the ruby laser in the treatment of Ota's nevus previously treated using other therapeutic modalities. AB - Ruby laser treatment, especially with a Q-switched laser, is remarkably effective for Ota's nevus, although a wide variety of other therapeutic modalities have had limited success. Consequently, laser treatment is now considered the treatment of choice. However, for Ota's nevus previously treated with dry ice cryotherapy (carbon dioxide snow), dermabrasion, free skin grafting, or other methods, therapy is still a challenge, even with the ruby laser. In this study, 14 patients with Ota's nevus previously treated using other modalities were treated using a Q-switched ruby laser. Eight patients previously underwent dry ice cryotherapy, three patients underwent free skin grafting, two patients underwent dermabrasion, and one patient received a cosmetic tattoo. The study group was composed of five male and nine female patients. The ages of the patients at the start of treatment ranged from 5 to 62 years. We concluded, based on the findings of this study, that Q-switched ruby laser therapy can provide favorable results even with lesions previously treated by other therapeutic modalities, provided that the treatment sessions are repeated more frequently and over a longer period of time than those used for untreated lesions and that they are combined with plastic surgical techniques such as scar resection or local flaps. PMID- 9858170 TI - The distally based superficial sural artery island flap: clinical experiences and modifications. AB - The coverage of defects of the Achilles tendon, malleoli, and heel remains a challenge to reconstructive surgeons. The distally based superficial sural artery island flap is vascularized by the median superficial sural artery, posterolateral septal perforators originating from the peroneal artery, neurovascular arteries of the sural nerve, and combinations of these systems as suprafascial plexus. We used distally based superficial sural artery island flaps for the reconstruction of defects of ankle, malleolus, and heel in 17 patients between 1991 and 1997. The largest flap we have used until today was 12 cm in width and 15 cm in length. All flaps have survived. However, we observed venous congestion and edema in two flaps. Also, marginal necrosis occurred in two flaps for which we had not taken the sural nerve with the flap. After these complications, we made some modifications. We left a skin extension over the fasciovascular pedicle and used it as a roof of the tunnel. We took the sural nerve and the deep fascia in all cases. The main advantage of this flap is a constant and reliable blood supply without sacrifice of a major artery. In addition, this is a one-stage, safe and easy procedure that can be used for large defects. PMID- 9858171 TI - Arterialized venous dorsal digital island flap for fingertip reconstruction. AB - Fingertip resurfacing is a challenging reconstructive problem; despite the existence of many different surgical methods, enthusiasm on developing versatile flap techniques has been continuing. In this report, we describe an arterialized venous dorsal digital island flap for fingertip reconstruction. The vascularity of the reverse dorsal digital island flap is augmented by performing an arteriovenous anastomosis between a dorsal vein in the flap and one of the proper digital arteries at the fingertip. Eight flaps were used in eight patients for the reconstruction of their fingertip defects. The ages of the patients ranged from 19 to 41 years, with an average of 28.4 years. In two cases, the flap was used as a neurosensorial flap by coapting the dorsal digital radial nerve to the digital nerve. The mean follow-up was 11 months, and all but one flap totally survived. The operative technique is easy except for the necessity of performing standard microvascular surgery. The flap may be a good alternative for repairing fingertip defects in selected cases. PMID- 9858172 TI - Correlation of clinical history and electrodiagnostic abnormalities with outcome after surgery for carpal tunnel syndrome. AB - A retrospective study on 294 wrists in 154 patients who had been diagnosed with carpal tunnel syndrome and subsequently had surgery performed was undertaken; both clinical and electrodiagnostic findings were correlated. The cases were divided into three groups based on electromyographic severity (mild, moderate, severe), and recovery from symptoms was evaluated after 1 week, 3 months, and 1 year. The cases were also divided into five groups based on symptom duration, and the same investigations were performed. All operations were conducted by applying the open release method with the limited-palmar incision technique. Operative outcomes showed no association between recovery from symptoms and the severity of electromyographic findings or the duration of symptoms, although the group that had the shortest duration of symptoms recovered faster than the long-duration groups statistically. Postoperative results after 1 year were also successful for those patients who had had symptoms of long duration. Of the 294 wrist operations studied, good to excellent postoperative outcomes were recorded in 242 cases (82 percent), fair outcomes in 39 cases (13 percent), and poor outcomes in 13 cases (4 percent). Patients whose electromyogram revealed double crush syndrome still showed improvement, with good-to-excellent results in 11 out of 15 cases (73 percent). Patients with diabetes mellitus also showed improvement, with good-to excellent results in 14 out of 19 patients (74 percent). This study showed that postoperative results were satisfactory within 1 year, regardless of the degree of electromyographic severity, symptom duration, presence of diabetes mellitus, or double crush syndrome. PMID- 9858173 TI - Multimodality evaluation of pressure relief surfaces. AB - Multimodality evaluation of six different support surfaces was performed measuring interface pressure, transcutaneous oxygen tension, and blood flow in pressure ulcer-prone areas on healthy subjects. Interface pressure was measured with a flexible force-sensing resistor array. The interface pressure distribution patterns of 10 healthy male volunteers were measured in the supine and lateral positions using a pressure sensor (force sensing resistor) array. Transcutaneous oxygen (TcPO2) and laser Doppler flowmeter probes were placed over the sacrum, the right trochanter, and the right ischium. Data were recorded for 15 minutes each with the subject in the supine and lateral decubitus positions. Statistically significant differences (p < 0.05) between the various surfaces could be observed for blood flow and pressure measurements, especially in the lateral position over the trochanter and for the TcPO2 measurements in the supine position over the sacrum. We conclude that measuring interface pressure, transcutaneous oxygen tension, and blood flow allows a more thorough evaluation of the physiologic effects of special support surfaces than any single technique. Future studies on patients will assess algorithms for using these measurement techniques to predict the efficacy of various support surfaces in minimizing pressure ulceration. PMID- 9858174 TI - Effect of tissue-plasminogen activator on leukocyte-endothelial interactions at the microcirculatory level. AB - In free tissue transfer and replantation surgery, there is a debate over whether any pharmacologic agents should be used to improve vessel patency and tissue survival. Because tissue-plasminogen activator (t-PA) is a highly effective and safe fibrinolytic, it may be useful in obtaining and maintaining vessel patency. The direct effects of t-PA on skeletal muscle hemodynamics and leukocyte activation at the microcirculatory level were investigated. Male Sprague-Dawley rats (n = 20) were divided into three experimental groups: control (n = 8), vehicle (n = 6), and t-PA (n = 6). Using the cremaster muscle flap model and intravital microscopy, red blood cell velocity, vessel diameter, capillary perfusion, endothelial edema index, and leukocyte-endothelial interactions (rolling, adhering, and transmigrating leukocytes) in postcapillary venules were measured. In the vehicle and t-PA groups, vehicle or t-PA was infused by means of a catheter inserted into the lower abdominal aorta for local infusion. Except for a significant reduction in the diameter of the first order arterioles from 117 microm to 82 microm (medians; p = 0.026), t-PA did not significantly affect red blood cell velocity, vessel diameter, or capillary perfusion compared with vehicle. However, leukocyte-endothelial interactions did differ significantly in postcapillary venules. Adhering leukocytes counted per visual field decreased from 4.67 in the vehicle group and 3.50 in the control group to 1.67 in the t-PA group (medians; p = 0.015 and p = 0.005, respectively); transmigrating leukocytes in the t-PA group decreased from 4.75 in the vehicle group and 3.50 in the control group to 1.67 in the t-PA group (medians; p = 0.002 and p = 0.043, respectively). t-PA treatment significantly decreased the number of both adhering and transmigrating leukocytes. These novel findings on leukocyte-endothelial interactions suggest that t-PA has anti-inflammatory effect. PMID- 9858175 TI - Charged beads enhance cutaneous wound healing in rhesus non-human primates. AB - Enhanced cutaneous wound healing by positively charged cross-linked diethylaminoethyl dextran beads (CLDD) was studied in a standardized incisional wound model in 20 adult and 20 geriatric Macaca mulatta (rhesus) partitioned equally over five time periods. Physiologic saline served as a control. Soft tissue linear incisions were prepared between and 1 cm inferior to the scapulae. There were four incisions per rhesus; each incision was 1.5 cm long with 1 cm of undisturbed tissue between incisions, and both the experimental CLDD and physiologic saline treatments were administered to each rhesus. The incision treatments were either CLDD and soft-tissue closure with 4-0 BioSyn sutures or sterile physiologic saline and closure with 4-0 BioSyn sutures. The hypothesis was CLDD would enhance cutaneous wound repair. Verification of the hypothesis consisted of clinical examinations and histologic and tensiometric evaluations on biopsy specimens at 10 and 15 days, whereas 5-day and 2- and 4-month groups were assessed clinically and biopsy specimens were assessed histologically. The clinical course of healing for all groups was unremarkable. At 10 days, incisions in adult rhesus treated with CLDD had a 30-percent greater tensile strength compared with the physiologic saline-treated incisions (p = 0.01), whereas for geriatric rhesus, the CLDD treatment proved to be 15 percent greater in tensile strength compared with the physiologic saline cohort (p = 0.11). By day 15, incisions in adult rhesus were 26 percent stronger than the saline treatment group (p = 0.07), and the difference was 36 percent (p = 0.02) for the geriatric rhesus. From 5 through 15 days, histologic observations revealed a gradual decrease in quantity and integrity of CLDD, with no remnants of CLDD at either 2 or 4 months. Macrophages and multinucleated giant cells were localized in the dermis and were associated with the CLDD. These cells decreased commensurately with the decrease of CLDD beads. The data suggest that CLDD can enhance significantly the tensile properties of healing cutaneous wounds in both adult and geriatric rhesus. Moreover, if the wound healing is enhanced in geriatric patients, this finding may be clinically germane to conditions where wound healing is compromised, such as in diabetics and patients on steroids. PMID- 9858176 TI - Promotion of second intention wound healing by emu oil lotion: comparative results with furasin, polysporin, and cortisone. AB - Previous studies showed that twice-daily application of emu oil lotion (mixture of emu oil/fat, vitamin E, and botanical oil) immediately after creation of full thickness skin defects delayed wound healing 6 days later, perhaps owing to its antiinflammatory actions. If administration was delayed for 48 hours, a two-fold promotion of wound contraction, epithelialization, and infiltration of organized granulation tissue was observed. In the present study, emu oil lotion was applied to full-thickness skin defects in rodents 24 hours after surgery. Six days postoperatively, wound contraction and infiltration of fronts of epithelialized and granulation tissue were assessed. Results indicated a two-fold promotion of all of the above parameters with emu oil lotion. No such effects were exerted by pure emu oil, furasin, cortaid, or polysporin. Data obtained indicate promise for emu oil lotion as an aid in treating full-thickness skin defects if applied after the major postinflammatory stages of wound healing have transpired. PMID- 9858177 TI - Single versus double arterial anastomoses in combined second- and third-toe transplantation. AB - Combined second and third toe transplantation is one good option for reconstruction of multiple digit amputation. However, the use of one or two arteries for pedicle anastomoses, which may influence the vascular complication and success rate, has never been addressed in the literature. This study includes a retrospective review of 57 combined second and third toe transplantation in 54 patients performed from February of 1983 through December of 1996. Group I, composed of 41 transplantations, underwent one arterial anastomosis, and group II, composed of 16 transplantations, underwent double arterial anastomoses during surgery if there were two recipient arteries available or whenever the second and third toes showed inadequate blood perfusion after one arterial anastomosis. In group I, 10 transplantations (24.4 percent) required re-exploration with a success rate of 92.7 percent (38 out of 41 transplantations). In group II, only one transplantation (6.2 percent) required re-exploration with successful flap salvage. The success rate was 100 percent for group II. Because the re exploration and success rates between groups I and II were not statistically significant according to two-tailed Fisher's exact test, the combined second and third toe transplantation is a reliable procedure using either single or double arterial anastomoses. PMID- 9858178 TI - Salvage reconstruction of an extensive facial deformity due to congenital giant hairy nevus. PMID- 9858179 TI - Functional alveolar ridge reconstruction with prefabricated iliac crest free flap and osseointegrated implants after hemimaxillectomy. AB - We achieved functional alveolar ridge reconstruction after hemimaxillectomy using a prefabricated iliac crest flap. The iliac crest was vascularized secondarily by a long rectus abdominis muscle flap with its inferior epigastric vessels intact to obtain an ideal anatomic location between the maxillary defect and microvascular anastomosis site. The iliac crest was tightly resurfaced with a split-thickness skin graft as well. After a bony surgical delay, the prefabricated iliac crest flap was microsurgically transferred to the face. Three osseointegrated implants were placed in the prefabricated iliac crest, and a dental prosthesis was worn with immobilization and stability. Our procedure enabled recovery of a satisfactory facial appearance and excellent masticatory function. PMID- 9858180 TI - Salvage of externally exposed ventricular assist devices. AB - Although externally exposed ventricular assist devices are associated with extremely high mortality rates, salvage may be accomplished by early aggressive wound debridement, transposition of a well-perfused autologous tissue (such as omentum), dead space obliteration, and adequate external coverage using vascularized tissue. The temporary suppression rather than the total eradication of the infection should be the goal of these procedures. PMID- 9858181 TI - Sequential transfer of free flap to two defects. AB - This case report describes the use of a single lateral arm flap sequentially transferred to two defects in a bilateral hand injury. We believe this is a novel approach in reconstructing defects in bilateral hands when a staged reconstruction is planned. PMID- 9858182 TI - A virtual reality system for bone fragment positioning in multisegment craniofacial surgical procedures. AB - This article reports our clinical experience since 1994 with rigid-motion tracking of bone fragments during craniofacial surgical procedures, using a virtual reality approach. Three noncollinear infrared diodes are fixed to the skull base. A pointer is used to register anatomic features on the patient to those on the computerized tomography-based model of the patient within a computer work station. Three diodes are then attached to each fragment just before the osteotomy is completed. Rigid motions of the fragment are thus tracked and reported to the surgeon by using virtual reality techniques. Errors in fragment positioning are reported both graphically and numerically with respect to a precomputed optimum fragment position. This guidance system allows multisegment midface osteotomies to be performed more precisely. The main problems encountered so far have been devascularization-infection and difficulties in maintaining correct position during application of rigid fixation. Devascularization infection problems have been addressed by minimizing surgical exposure of the bone. Soft-fixation plates and temporary Kirschner wire fixation have helped with intermediate positioning, but an intraoperative mechanical positioning device would be useful in the future. PMID- 9858183 TI - A simple technique for reconstruction of the umbilicus, using two twisted flaps. AB - A successful umbilical reconstruction is described, using two twisted flaps with one pedicle. This technique is easy and simultaneously revises the scar of the abdominal wall. With this procedure, the umbilicus has a natural appearance with sufficient depression and normal-appearing wrinkles. PMID- 9858184 TI - Method for securing a subcuticular suture with minimal buried knot. AB - Subcuticular, intradermal suturing with retained suture has been performed with a variety of techniques requiring bulky buried knots. This report outlines a new method for closing a skin wound under light to moderate tension with fewer problems of buried knot bulkiness and gapping of the wound ends. The suture method was developed in a porcine model and has been used in clinical suturing with 3-0, 4-0, and 5-0 suture material. Outcomes for 23 cases have been satisfactory with no dehiscences and no exposure of knot material. This technique provides the surgeon with an excellent way to approximate skin wounds with a retained sucuticular suture. A modified technique for small 3- to 10-mm wounds is also described. PMID- 9858185 TI - Endoscopic omental harvest. AB - An omental flap is useful in reconstructive surgery, but harvesting such a flap generally requires laparotomy. However, endoscopic surgery facilitates harvesting an omental flap without open laparotomy. We performed endoscopic omental harvest in two patients. We described the procedure of endoscopic omental harvest, which is different from that reported previously. Four access ports were required: two placed lateral to the right rectus margin, one placed lateral to the left rectus margin, and one placed in the infraumbilical area. The stomach was suspended from the peritoneum for the dissection of the gastric rami. The vessels from the gastroepiploic arcade to the greater curvature of the stomach were individually clipped and divided. The omentum then was dissected to the transverse colon and the lower portion of the omentum was dissected along the transverse colon. Finally, the right side of the omentum was dissected. The omentum was transferred using the right gastroepiploic vessels for anastomosis. The advantages of endoscopic harvest are an inconspicuous scar, minimal operative pain, and early recovery. The disadvantages include a long procedure time. At the present time, endoscopic harvest of the left side of the omentum is problematic because of difficulty in identifying the left omental artery and the risk of injury to the spleen. However, these limitations will likely be resolved in the future. PMID- 9858186 TI - Anchoring galeal flaps for scalp reduction procedures. AB - This article describes an operative technique, based on the use of three anchoring galeal flaps, aimed at reducing the percentage of "stretch-back" that occurs after performing scalp reduction procedures. In 12 male patients undergoing a midline scalp reduction procedure, three rectangular (2 x 3 cm) galeal flaps in direct continuity with the longitudinal margin of the left scalp flap were sutured individually to the galeal undersurface of the right scalp flap to draw the two scalp flaps toward the midline of the scalp and to relieve the wound margins of closing tension. Tattoo marks were placed on the patient's scalp at the level of the vertical lines drawn through the external auditory meatuses (A1-A2) and 6 cm more posterior (B1-B2) to measure the movement and stretching of the scalp. The results were compared with those obtained from a control group of 13 male patients who underwent the same surgical procedure but without the use of the anchoring galeal flaps. Mean stretch-back (as measured 4 weeks postoperatively) at level A1-A2 was 8.3 mm in the control group and 1.6 mm in the experimental group. The mean stretch-back at level B1-B2 was 7.7 mm in the control group and 0.9 mm in the experimental group. A statistically significant difference (p < 0.005) was found between data from the control and experimental groups regarding the above-reported stretch-back values at both levels. The use of the described galeal flaps allowed us to obtain an 80.93-percent and an 88.09 percent stretch-back reduction at levels A1-A2 and B1-B2, respectively, 1 month postoperatively. PMID- 9858187 TI - Lower eyelid hernia repair for palpebral bags: a comparative study. AB - The standard treatment for herniated "bags" of the lower eyelid is surgical removal of excess fat. Sachs and Bosniak in 1986 and de la Plaza and Arroyo in 1988 described a new technique for treatment of palpebral bags that consisted of returning the herniated fat to the orbital cavity and retaining it by continuous sutures of the capsulopalpebral fascia either to the dehiscent portion of the orbital septum or to the periosteum of the lower orbital rim. This article reports a prospective study of 26 patients who underwent standard blepharoplasty in one lower eyelid and capsulopalpebral fascia hernia repair in the other lower eyelid. All were evaluated at 6 weeks and at 6 months after surgery, and the outcomes were compared. The results of the two different techniques in the same patient have shown comparable aesthetic outcomes in the treatment of palpebral bags. However, results indicate that the capsulopalpebral fascia hernia repair technique carries less discomfort and pain during the operation and may be less prone to postoperative bleeding and hematoma formation. In addition, in contrast to standard lower blepharoplasty with fat resection, hollowing of the lower lid or potential sunken appearance of the globe may remain absent with capsulopalpebral fascia hernia repair beyond the 6-month period of this study. PMID- 9858188 TI - Computerized morphometric quantitation of elastin and collagen in SMAS and facial skin and the possible role of fat cells in SMAS viscoelastic properties. AB - Recently, the superficial musculoaponeurotic system (SMAS) was found to be a composite tissue comprising collagen, elastic fibers, and fat cells in an extracellular viscous matrix. Both SMAS and facial skin tissues exhibit viscoelastic properties, but SMAS tissue has delayed stress relaxation. As a consequence, SMAS is viewed as a firmer elastic foundation for the more viscous facial skin. In some patients, a slackening effect of SMAS tissue takes place over a period ranging from weeks to months after tightening. To determine the relative quantity of viscoelastic components and better understand their biomechanical behavior, a quantitative morphometric study of the elastic and collagen fibers in the SMAS and facial skin was conducted. Thirty-four SMAS preparations were taken from 17 patients during either primary face lift operations (12 women) or reoperative face lift procedures (4 women, 1 man), which were performed 4 to 9 months after the original surgery, to examine the elastin and collagen content. For comparison, preauricular skin was also gathered from these patients. The specimens were stained with Weigert's staining to identify elastin and collagen fibers. Using a computerized morphometric analysis, 100 fields of each SMAS and skin specimen were examined. According to our findings, the average percentage of elastin and collagen fibers in SMAS and facial skin was as follows: (1) the percentage of elastin fibers in the SMAS was 4.71 +/- 1.2 (standard error of mean +/- 0.0291); (2) the percentage of elastin fibers in the skin was 6.1 +/- 1.8 (standard error of mean +/- 0.0436); (3) The percentage of collagen fibers in the SMAS was 38.7 +/- 5.9 (standard error of mean +/- 0.1430); and (4) the percentage of collagen fibers in the skin was 48.47 +/- 6.96 (standard error of mean +/- 0.1688). A statistical significance of p < 0.0001 was demonstrated between the collagen and elastin groups. A different percentage of elastin and collagen fibers was found among the 17 patients and within each of them separately. Neither gender nor age differences were found regarding elastin and collagen fiber content. No statistical differences were demonstrated between specimen sources, i.e., whether the operations were primary or reoperative face lift procedures. Findings from previous studies indicate that the cheek has two viscoelastic layers, the skin and the SMAS. The proportional similarity in average percentages of elastin and collagen in SMAS and facial skin cannot explain the relatively delayed stress relaxation effect of the SMAS. Therefore, the fat cells that are found exclusively in the SMAS probably lend a certain degree of firmness to this layer and play a significant role in the long-term efficacy of SMAS surgery. PMID- 9858189 TI - Redraping the inferior orbicularis arc. AB - The surgical technique of redraping of the inferior arc of the orbicularis oculi muscle is used primarily to produce lower lid and midfacial smoothing in patients undergoing aesthetic surgery. The midfacial fat compartments, suborbicularis oculi fat and malar fat, are bound to the orbicularis muscle by the superficial muscular aponeurotic system so that redraping the orbicularis muscle also repositions the midface. Orbicularis arc redraping should be accompanied by lateral canthoplasty to ensure stability to the shape of the eyelid fissure postoperatively. Modifications in orbicularis redraping and canthoplasty technique are necessary in patients with prominent eyes and distensible lower lids. Supraplacement of canthal fixation is needed in patients with prominent eyes, and lid shortening is needed in patients who have distensible lower lids. This technique also can be used in patients undergoing reconstructive surgery for correction of lower lid retraction because of its ability to recruit periorbital skin upward into the lower lid. For more severe cases of lower lid retraction after aesthetic surgery, adjunctive procedures such as spacer implants in the lower lid and periosteal flap canthoplasty can be used together with the orbicularis arc redraping to rehabilitate patients. PMID- 9858190 TI - Simultaneous face lifting and skin resurfacing. AB - Although face lifting and skin resurfacing both produce dramatic facial rejuvenations, the simultaneous combination has never been popularized. The development of new methods of lifting and resurfacing may now allow a reevaluation of this combination. The objective was to evaluate the safety and efficacy of this combined face lift and skin resurfacing. The facial skin was resurfaced with a trichloroacetic acid peel or a "short-pulse" CO2 laser and, then, lifted after liposuction and superficial musculoaponeurotic system tightening. A dramatic rejuvenation was produced in these 25 cases. There were no incidents of persistent erythema, pigmentation, hyperpigmentation, or full thickness flap necrosis. Side effects were the usual sequelae of edema and ecchymoses. The small areas of "dusky" erythema of the skin flaps were no more significant than those after a face lift alone. By following the guidelines developed in this report, the combination of controlled skin resurfacing with face lifting is safe and effective. Other procedures, such as blepharoplasties, can also be added. After one recovery period of 6 to 8 weeks, these patients reported that they looked 15 to 20 years younger. The complexion continued to improve during the 1-year follow-up period. PMID- 9858191 TI - The ideal dorsal graft: a continuing quest. PMID- 9858192 TI - Tissue preservation in hand injury: the first step to toe-to-hand transplantation. AB - The recommendations outlined in this editorial for the initial management of nonreplantable amputation injuries emphasize tissue preservation. This facilitates future toe-to-hand transplantation in terms of reconstructive options, functional and aesthetic outcomes in the hand, and donor-site morbidity in the foot. It is hoped that surgeons involved in the emergency surgical care of such cases will find these guidelines useful. Although tissue conservation is the main goal at this stage of treatment, this should not be at the expense of tissue viability. If in doubt about management issues, advice should be sought from the reconstructive microsurgery unit. PMID- 9858193 TI - A simple method for tunneling a nerve graft during facial reanimation. PMID- 9858194 TI - Use of Sorbsan, a calcium alginate fiber dressing, as a drug reservoir in iontophoretic transdermal delivery. PMID- 9858195 TI - Tulip flap for reconstruction of the central tubercle in cleft lip patients. PMID- 9858196 TI - Direct effect of vitamins at the time of palatal fusion. PMID- 9858197 TI - The extended supraplatysmal plane face lift. PMID- 9858198 TI - Reverse flow in microsurgery. PMID- 9858199 TI - Tensor fasciae latae in abdominal wall reconstruction. PMID- 9858200 TI - Use of an existing tattoo to conceal a surgical incision. PMID- 9858201 TI - Intravenous fluids and bupivacaine are contraindicated in tumescent liposuction. PMID- 9858202 TI - Adiposubcutaneous flap for reconstruction of first web space depression in ulnar paralysis in leprosy patients. PMID- 9858203 TI - We have missed the essential step of treatment for plantar ulcers in leprosy patients for too long. PMID- 9858204 TI - Microsurgery in angioacces gaining for chronic dialysis is a benefit for the patient and a valuable teaching instrument. PMID- 9858205 TI - Telomeres and haemopoiesis. PMID- 9858206 TI - Developments in allogeneic peripheral blood progenitor cell transplantation. PMID- 9858207 TI - Composition and function of peripheral blood stem and progenitor cell harvests from patients with severe active rheumatoid arthritis. AB - High-dose chemotherapy with autologous stem cell rescue has been proposed as an intensive therapy for severe rheumatoid arthritis (RA). In view of previous observations of abnormal haemopoiesis in RA patients, the composition and function of peripheral blood stem cell harvests (PBSCH) was investigated. Compared with PBSCH from healthy allogeneic donors mobilized with the same dose of G-CSF (filgrastim; 10 microg/kg/d, n = 14), RA PBSCH (n = 9) contained significantly fewer mononuclear cells (375 v 569 x 10(6)/kg, P = 0.03) and CD34+ cells (2.7 v 5.8 x 10(6)/kg, P = 0.003). However, there were increased proportions of CD14+ cells (P = 0.006) and CD14+ CD15+ cells (the phenotype of previously described 'abnormal' myeloid cells, P = 0.002) in the RA PBSCH which translated into 3.5- and 7-fold increases respectively on a per CD34+ cell basis. There were no differences in T-cell activation status as judged by proportions of CD4+ and CD8+ expressing CD45RA, CD45RO, HLA-DR and CD28 (RA PBSCH, n = 7, donor PBSCH, n = 5, P = 0.2-0.7). Phytohaemagglutinin responses determined fluorocytometrically with induction of CD69 expression were reduced in CD4+ and CD8+ cells following filgrastim administration in 3/3 RA patients tested. Compared with bone marrow as a potential source of CD34+ cells, PBSCH contained 11-fold more T cells (P < 0.0005), 8-fold more B cells (P < 0.0005) and 4-fold more monocytes (P = 0.02). In short-term methylcellulose culture there were no differences in colony counts (CFU-GM, CFU-GEMM, BFU-E) per CD34+ cell from PBSCH from RA patients (n = 11) and healthy donors (n = 10). Long-term culture initiator cells were cultured successfully from cryopreserved PBSCH from RA patients (n = 9). In conclusion, PBSCH from RA patients differed significantly in composition from normal individuals, but in vitro studies support normal stem and progenitor cell function. Changes in T-cell function occur during mobilization in RA patients. This work provides reassurance for the use of PBSCH as haematological rescue and baseline data for clinical trials of graft manipulation strategies in patients with RA. PMID- 9858209 TI - Extensive phenotypic analysis of CD34 subsets in successive collections of mobilized peripheral blood progenitors. AB - The transplantation of mobilized progenitor cells after high-dose chemotherapy shortens haemopoietic engraftment. CD34 cell subsets were examined in 20 consecutive mobilized progenitor cell collections obtained from patients with solid tumours that had not been previously treated. The analysis of CD34 cells was based on the expression of intracellular antigens, surface antigens including CD38, and cell size using multi-dimensional flow cytometry. We also correlated the numbers of stem cell subsets reinfused to haemopoietic recovery. The majority of CD34+ cells expressed CD13 and CD33. A significant proportion was cytoplasmic myeloperoxidase (cMPO) positive. CD34+ MPO+ cells increased significantly in late collections. MPO expression was related to cell size. Cells expressing CD13 also increased in late collections in parallel to CFU-GM count. Small subpopulations of CD34+ CD38+ were committed to B cells, T cells and erythroid cell lineages. A small population expressing the megakaryocytic antigen had a small size and were predominantly CD38-. A minor subpopulation expressed stem cells antigens. These were significantly higher in late collections (CD34+ Thy-1+ and CD34+ CD33-). After mobilization, patients received three cycles of intensive chemotherapy followed by reinfusion of mobilized progenitors (5.45 x 10(6)/kg CD34+ cells, range 3.4-11.88). The numbers of reinfused CD34 cells or the individual subsets did not influence recovery of leucocytes (9 d) or platelets (9 d). In conclusion, the numbers of stem cells and their subsets differed between collections and, in unpretreated patients receiving intensive chemotherapy, there was no delayed engraftment when sufficient numbers of stem cells were reinfused. The recovery period was short and not correlated to any stem cell subsets. PMID- 9858208 TI - Evaluation of breast tumour cell contamination in the bone marrow and leukapheresis collections by RT-PCR for cytokeratin-19 mRNA. AB - There is considerable interest in an autologous transplantation (AT) programme for patients with high-risk breast cancer; however, the issue of the incidence of occult bone marrow (BM) micrometastasis at diagnosis, and the cancer contamination of peripheral blood stem cell (PBSC) collections used for haematological rescue, is still debated. The presence of BM micrometastasis was evaluated in bilateral BM biopsies obtained at diagnosis of 33 patients with stage II/IIIA breast cancer using: (i) a 'nested' reverse transcriptase polymerase chain reaction (RT-PCR) assay for cytokeratin 19 (K19) mRNA, (ii) histology, and (iii) immunohistochemistry (IHC) analysis with a panel of three monoclonal antibodies. The RT-PCR assay only was used to determine contamination of PBSC collections obtained after priming with recombinant human granulocyte colony stimulating factor (rhG-CSF). K19 transcripts in one or both BM samples were detected in 48% of patients at diagnosis, with an overall 85% concordance with the results of IHC analysis. On the other hand, 56% of PCR- and IHC-positive BM samples were diagnosed as 'normal' on histological analysis. 57% of patients showed K19 mRNA in at least one PBSC collection; the possibility to have contaminated PBSC collections was significantly higher in patients with K19 positivity in BM at diagnosis. In four patients who had shown K19 positivity in BM and in PBSC collections, immunoselected CD34+ cells used for haematological rescue were K19-negative. There was a trend towards longer relapse free survival (RFS) in patients transplanted with K19-negative PBSC collections as compared to the others. In conclusion, a substantial proportion of patients with high-risk non-metastatic breast cancer present occult BM micrometastasis at diagnosis and also show cancer contamination of PBSC collections used for AT. These might represent a category of patients with poorer prognosis after AT, and possible candidates for more intensive and/or alternative therapeutic regimens, including AT with purged PBSCs. PMID- 9858210 TI - Bone marrow transplantation for adults with acute leukaemia and 11q23 chromosomal abnormalities. AB - Adults with acute leukaemia and abnormalities of chromosome 11q23 have a poor prognosis when treated with conventional chemotherapy. To determine whether more intensive therapy can improve outcome for patients with this karyotypic finding, a retrospective analysis of all patients with acute leukaemia and 11q23 abnormalities treated at our centre was performed. 12 patients were treated with conventional chemotherapy alone (CC); 20 patients received high-dose chemo/radiotherapy (HDCT) with autologous (seven patients) or allogeneic (13 patients) bone marrow transplantation (BMT). The treatment-related mortality was 25% [95% Confidence Interval (CI) 7-69%] for the CC group and 46% (CI 25-73%) for the BMT group (P = 0.69). Cumulative risk of leukaemia progression was 89% (CI 61 100%) in the CC patients and 38% (CI 12-69%) in the BMT patients (P = 0.001). The 2-year event-free survival for patients treated with CC was 8% (CI 0-31%) and for patients receiving HDCT and BMT was 34% (CI 14-54%) (P = 0.03). These results confirm that conventional chemotherapy is rarely curative for adults with acute leukaemia and 11q23 abnormalities but that HDCT with BMT can result in long-term survival in a significant proportion of patients. PMID- 9858211 TI - Expansion of megakaryocyte progenitors from human umbilical cord blood using a new two-step separation procedure. AB - Cord blood (CB) transplantation is primarily performed in children, rather than in adults, due to the low number of haemopoietic progenitor cells obtained from the small volume of a single CB collection. Prolonged thrombocytopenia is a major problem following CB transplantation. Efforts are currently underway to expand the number of CB progenitor cells ex vivo, in order to enable transplantation in adults and to decrease the period of thrombocytopenia. In this study we investigated different techniques for enrichment and expansion of megakaryocyte (Mk) progenitor cells and haemopoietic stem cells from CB. CBs from 20 normal deliveries were depleted of red blood cells (RBC) by dividing each sample and testing cell separation on 3% gelatin, Hespan, Ficoll-Paque or a two-step 3% gelatin followed by Ficoll-Paque separation. The two-step procedure was found to be superior to the other methods in enrichment of the Mk progenitor cells (CFU Mk) (34.3-fold), while at the same time retaining the number of myeloid and erythroid progenitors, CD34+ and CD41+ cells. In short-term (14d) liquid culture of non-adherent nucleated cells isolated by gelatin and Ficoll-Paque, a 40-fold expansion of clonable Mk progenitor cells was obtained in the presence of thrombopoietin (r-hu-TPO) and stem cell factor (r-hu-sCF. In similar cultures of isolated CD34+ cells, a 100-fold clonable Mk progenitor was obtained at day 14. Therefore this new technique may facilitate the ex vivo expansion of Mk progenitor cells and be adopted for future use in CB transplantation. PMID- 9858212 TI - Autologous bone marrow transplantation for non-Hodgkin's lymphoma resulting in long-term remission of coincidental Crohn's disease. AB - Crohn's disease usually runs a chronic relapsing and remitting course, characterized by a repeated need for immunosuppressive drug therapy or often surgery. and is considered to be incurable by standard treatment. We report a case of successful long-term disease control of Crohn's disease following autologous bone marrow transplantation. This case provides further support for the concept that some forms of severe immune-mediated diseases may be amenable to treatment by high-dose therapy with autologous stem cell support. PMID- 9858213 TI - Seasonal variation in the incidence of Hodgkin's disease. AB - Earlier literature suggested there may be a seasonal rhythm of onset of Hodgkin's disease. This issue has been re-examined using population-based prospectively collected data with high ascertainment levels. The Data Collection Study (DCS) of the Leukaemia Research Fund (LRF) Centre for Clinical Epidemiology (University of Leeds) generated the information used, which was based on a population of 13.5 million--about one quarter of England and Wales--over 10 years. The RYE histopathological classification was employed. The findings show that in patients with nodular sclerosing histopathology there was a highly significant circannual rhythm with a low amplitude (extent of seasonal variation) and a peak in March. A significant, but different, rhythm with a high amplitude and a peak in August was found in lymphocyte predominant Hodgkin's disease. However, this finding is less certain, due to smaller numbers and a lower significance level. The main conclusion is that there is a highly significant seasonality in nodular sclerosing Hodgkin's disease. The findings provide further evidence that nodular sclerosing and lymphocyte predominant may be two different diseases. The differing seasonality rhythms may provide aetiological clues. PMID- 9858214 TI - In-vivo immune responses to idiotypic VH complementarity-determining region 3 peptide vaccination in B-cell non-Hodgkin's lymphoma. AB - Idiotypic IgM expressed by B-cell non-Hodgkin's lymphoma (NHL) is clone specific and an ideal tumour antigen for immune targeting. We previously showed that repeated rounds of in vitro stimulation of peripheral blood mononuclear cells (PBMC) with autologous idiotypic heavy variable (VH) complementarity-determining region (CDR)-3 peptide produced T-cell lines able to proliferate, produce Th1 cytokines and kill autologous target cells pulsed with the CDR3 peptide. Furthermore, fresh autologous lymphoma cells were lysed by these T-cell lines, suggesting a potential for the clinical use of the CDR3 peptide for lymphoma vaccination. In this study, we determined the specific immune responses of a patient following vaccination with VH CDR3 peptide to determine if similar immune responses observed in vitro could be elicited in vivo. The patient received three fortnightly subcutaneous injections of idiotypic CDR3 peptide. The injection sites developed inflammation after the second and third vaccinations. Peripheral blood T cells were able to proliferate and produce IFN-gamma upon rechallenge with the CDR3 peptide in vitro. Cytotoxic T lymphocytes (CTLs) for autologous CD3 depleted CDR3-pulsed PBMC were detected, but only after the first vaccination. T cell lines generated after vaccination could lyse peptide-pulsed autologous target cells. We also detected the production of anti-CDR3 peptide antibodies in the patient's serum. We conclude that naked CDR3 peptide vaccination can result in the generation of potentially beneficial specific immune responses as predicted by in vitro studies. PMID- 9858215 TI - Establishment of an IL-2-dependent cell line derived from 'nasal-type' NK/T-cell lymphoma of CD2+, sCD3-, CD3epsilon+, CD56+ phenotype and associated with the Epstein-Barr virus. AB - A novel interleukin-2 (IL-2)-dependent cell line, HANK1, was established from a patient with CD56+ NK/T-cell lymphoma arising in the retroperitoneum. Morphologically, HANK1 is a pleomorphic large cell line with irregular nuclei, which contains azurophilic granules in the cytoplasm. Immunophenotypic analysis showed that HANK1 expressed CD2, CD3epsilon, CD56, TIA-1, granzyme B, and HLA-DR, but no other T-lineage markers. These features were the same as seen in the original tumour, and are highly characteristic of nasal and 'nasal-type' NK/T cell lymphoma as described in the proposed W.H.O. classification. Genotypically, this cell line also demonstrated the germline configuration of the T-cell receptor beta, gamma and the immunoglobulin heavy chain genes and clonal integration of the Epstein-Barr virus (EBV) together with antigen expression with a type II latency pattern (LMP-1+ and EBNA2-). Furthermore, Southern blot analysis using the EBV termini as probes confirmed its derivation from the original lymphoma, and revealed that it contained multiple copies of the EBV genome. Dose-dependent growth on IL-2 was observed in an in vitro study with a doubling time of 3 d at maximal stimulation. These data indicate that HANK1 seemed to preserve the biological characteristics of the original tumour and therefore may serve as a good model for the further analysis of unusual 'nasal type' NK/T-cell lymphoma. PMID- 9858216 TI - Paclitaxel plus high-dose cyclophosphamide with G-CSF support in patients with relapsed and refractory aggressive non-Hodgkin's lymphoma. AB - Based on the single-agent activity of both paclitaxel and cyclophosphamide in the treatment of non-Hodgkin's lymphoma (NHL), we conducted a phase II study to evaluate the efficacy of the combination of the two drugs in patients with refractory and relapsed aggressive NHL. All patients received 900 mg/m2 bolus of cyclophosphamide intravenously daily for 3 consecutive days with a concurrent infusion of 150 mg/m2 of paclitaxel over 72 h (50 mg/m2/d). 24 h after the completion of chemotherapy, patients received subcutaneous injections of 5 microg/kg of granulocyte-colony stimulating factor (G-CSF) daily until white cell count recovery. Treatment was repeated every 3 weeks. Patients who had at least a partial response (PR) after two courses continued to receive a maximum of four courses. Patients with responding disease were allowed to undergo high-dose chemotherapy followed by stem-cell/bone marrow transplantation if they were eligible. Of the 77 patients who were eligible for the study, 74 (96%) were evaluable for toxicity and treatment response. The overall response rate was 45% (95% CI 33-57%). Patients who received treatment after their disease relapsed from a complete response (CR) had an 81% response rate (38% CRs), whereas those with primary refractory disease had a 22% response rate. Toxicities of > grade 2 included alopecia (100%) and stomatitis (25%). Neutropenic fever of grade > 2 occurred after 18% of the courses, and platelet count of < or = 20 x 10(9)/l developed after 20% of the courses. Thus, the combination of paclitaxel plus high dose cyclophosphamide is an effective new regimen in the treatment of refractory and relapsed NHL. PMID- 9858217 TI - CD40 regulation of death domains containing receptors and their ligands on lymphoma B cells. AB - Within the tumour necrosis factor (TNF) family the induction of apoptosis is restricted to some ligand-receptors pairs, including TNF-TNF receptor type I (TNFRI/p55), FasL-Fas, TNF-related apoptosis-inducing ligand (TRAIL) and its death-receptors (DR)-4 and -5. The pair CD40L-CD40 belongs to the same family but rescues B cells from apoptosis. To investigate how these opposing actions are cross-linked, purified follicular lymphoma (FL) cells were activated upon a human CD40L-transfected murine fibroblastic layer, then RNA messengers for the above molecules were analysed using RT-PCR. The observed down-modulation of TRAIL and up-regulation of TNF and Fas transcripts might account for CD40-CD40L-mediated FL cell survival. PMID- 9858218 TI - Bolus administration of cladribine in the treatment of Waldenstrom macroglobulinaemia. AB - This phase II clinical trial evaluated bolus cladribine as a single agent in Waldenstrom macroglobulinaemia (WM). Cladribine was administered to 20 patients at a dose of 0.12 mg/kg/d by 2 h intravenous infusion for 5 consecutive days at monthly intervals for three courses. Partially responding patients were continued on therapy until maximal response and/or prohibitive toxicity, to a maximum of eight courses. Complete responders were treated with one additional course of cladribine. After a median of three courses of cladribine, all 20 patients were evaluable; one achieved a complete response (CR) (5%) and 10 achieved a partial response (PR) (50%). The median duration of response follow-up was 28 months (range 1-37 months). Four of 7 (57%) untreated and 7/13 (54%) previously treated patients responded. The major toxicity encountered was myelosuppression with 60% of patients demonstrating grade 3 or 4 neutropenia. Non-haematological toxicities included two patients with herpes zoster and two patients with non-melanoma skin cancers. At a median follow-up duration of 20 months, 17 patients remain alive and three have died. We confirm that bolus cladribine is an effective and safe method of drug delivery in WM patients. Recommendations regarding the equivalence of the continuous infusion and bolus methods in untreated patients requires further study. Bolus cladribine is more convenient and less costly than infusional cladribine since it obviates the need for central catheters and infusional devices. PMID- 9858219 TI - Anticipation in familial plasma cell dyscrasias. AB - Familial myeloma was described as early as 1925; however, the causative factors are unknown. Studies of families with other familial haematological malignancies demonstrate anticipation. Five new families are described in which plasma cell dyscrasias occurred in parent and child generations (six such pairs), and data were pooled with those of 16 other families (with 20 parent-child pairs affected) recorded in the literature. Disease-free survival for parent and child generations were each estimated and differences in the disease-free survival between generations were tested by the log-rank and signed rank methods. In all six previously unreported parent-child pairs with plasma cell dyscrasia and in 18/20 such pairs found in the literature, the disease occurred at an earlier age in the child generation. The median age of onset of myeloma in the parent and child generations of all 26 pairs was 71 years (95% CI 67-78 years) and 50 years (95% CI 45-55 years), respectively (P<0.0001). The ages of onset of malignant plasma cell dyscrasias in the parent and child generations of these families compared with patients in the general population were significantly different for the child generation (P<0.005) but not for the parent generation. It would appear that anticipation occurs in familial myeloma. PMID- 9858220 TI - Chromosomal abnormalities in systemic amyloidosis. AB - Primary systemic amyloidosis (AL) is a plasma cell disorder characterized by deposition of monoclonal light chains in different organ systems. Although multiple and complex numerical chromosomal abnormalities have been described in patients with multiple myeloma, it is currently unknown whether such changes occur in systemic amyloidosis. Bone marrow samples from 21 patients with AL were studied by standard cytogenetics and interphase fluorescence in situ hybridization (FISH) for the presence of numerical chromosomal abnormalities. We tested for six chromosomes (7, 11, 9, 15, 18 and X) using centromere-specific probes. The monoclonal plasma cells were identified by simultaneous fluorescent staining of the monotypic cytoplasmic immunoglobulin. We compared these results with those obtained from 19 patients with monoclonal gammopathy of undetermined significance (MGUS) and normal controls. Multiple numerical chromosomal abnormalities were detected in AL by interphase FISH, including trisomy of chromosomes 7 (42%), 9 (52%), 11 (47%), 15 (39%), 18 (33%) and X (13% in women and 54% in men). Monosomy of chromosome 18 was seen in 72% of cases. Previous exposure to alkylator therapy did not appear to correlate with these abnormalities. No significant difference was observed in the prevalence of these abnormalities between AL and MGUS. Multiple chromosomal numerical abnormalities were detected by interphase FISH analysis in patients with AL, especially monosomy of chromosome 18. Aneuploidy in the monotypic plasma supports a neoplastic nature for the disorder. PMID- 9858221 TI - Expression of p210 and p190 BCR-ABL due to alternative splicing in chronic myelogenous leukaemia. AB - The hallmark of chronic myelogenous leukaemia (CML) is the presence of the Philadelphia chromosome and its resultant fusion message, BCR-ABL, and fusion protein, p210. Patients with CML in blast crisis, or with Philadelphia positive acute lymphoblastic leukaemia (ALL), can have a smaller BCR-ABL fusion transcript possessing only the first exon of BCR fused to ABL. This smaller transcript encodes a 190 kD protein which is more strongly transforming than the p210 protein derived from the larger CML-associated transcript. We performed RT-PCR on samples from CML patients in chronic phase to determine the frequency and mechanism of p190 and p210 co-expression and to see if this correlated with clinical indices. We examined the peripheral blood or marrow of 67 patients with CML and found that 35 of them expressed both transcripts whereas the remainder expressed the p210-encoding transcript exclusively. Additional PCR products of an intermediate size were also frequently detected and have been isolated and sequenced. Data from two of these products indicate that they are the result of alternative splicing and include variable combinations of BCR exons. We believe that the expression of the p190-encoding transcript in the chronic phase of CML is also due to alternative splicing. A comparison of patients co-expressing the p190- and p210-encoding transcripts with those patients who expressed only the p210-encoding transcript detected significantly higher white blood cell (WBC) counts and blast cell counts at time of testing as well as significantly higher white blood cell counts at diagnosis. PMID- 9858222 TI - The susceptibility of Philadelphia chromosome positive cells to FAS-mediated apoptosis is not linked to the tyrosine kinase activity of BCR-ABL. AB - We investigated whether inhibition of the BCR-ABL tyrosine kinase by the CGP57418B compound would render chronic myeloid leukaemia (CML) cells susceptible to Fas (CD95, Apo-1)-mediated cell death. Only two (AR230 and SD1) out of 10 BCR ABL positive cell lines were found to express the CD95 protein. No change in Fas expression was observed in any of the 10 cell lines after 48 h exposure to CGP57418B. AR230 cells were resistant and SD1 cells were partially resistant to Fas-mediated apoptosis induced by ligation of the Fas receptor to an anti-Fas IgM antibody. Pre-incubation with 1 microM CGP57418B did not change the susceptibility of these cell lines to Fas-mediated cell death. Similar results were observed in experiments with CD34+ cells from CML patients and from normal individuals. The data suggest that, in contrast to some cytotoxic drugs, the CGP57148B tyrosine kinase inhibitor utilizes a pathway other than the CD95 system in order to induce apoptosis in CML cells. PMID- 9858223 TI - Arsenic trioxide inhibits growth of human T-cell leukaemia virus type I infected T-cell lines more effectively than retinoic acids. AB - Adult T-cell leukaemia (ATL) is difficult to cure using conventional therapies. Recently the therapeutic possibility of retinoic acids (RA) has been reported. In this study, suppression of in vitro growth of human T-cell leukaemia virus type I (HTLV-I) infected T-cell lines and fresh ATL cells by arsenic trioxide (As2O3) were evaluated by comparison with a series of RA derivatives. Proliferation of four HTLV-I-infected T-cell lines was significantly reduced within 72 h by 1.0 micromol/l As2O3. Growth of two out of four HTLV-I-infected T-cell lines was also inhibited by 1.0 micromol/l RA, but to a lesser extent than by As2O3. The mechanism of this growth inhibition was due to the induction of apoptosis. Apoptosis was also induced in fresh ATL cells from patients by AS2O3, but far less by RA. As described in patients with acute promyelocytic leukaemia, 1.0 micromol/l of As2O3 can be safely achieved in the serum of patients; however, it is difficult to maintain this concentration of RA. In conclusion, As2O3 has therapeutic potential for the treatment of ATL and may be far more clinically beneficial than RA. PMID- 9858224 TI - Ultrastructural characteristics and lysozyme content in hypergranular and variant type of acute promyelocytic leukaemia. AB - We investigated the electronmicroscopic (EM) features and cellular lysozyme (LZ) content in 16 cases of acute promyelocytic leukaemia (APL): 11 cases of the hypergranular form (M3) and five cases of the microgranular variant (M3-V). The main EM features in all cases were: irregular, folded or bilobed nuclei, many cytoplasmic granules, distended rough endoplasmic reticulum (RER) cisternae which, in some cases, presented as stellate forms (more frequent in M3-V), and bundles of cytoplasmic microfilaments. Many Auer rods were present in M3 cases and few in M3-V; most of these disclosed parallel tubular arrays (PTA) with a varied periodicity ranging from 13 to 26 nm. There was a significant difference between M3 and M3-V (P<0.0001) in both the number of granules per cell section (62.9 +/- 34.5 v 38.0 +/- 23.6) and in the granule section area (0.044 +/- 0.033 v 0.026 +/- 0.015 microm2). In some cases, mainly in M3-V, we found cells with large granules containing PTA which probably represent poorly developed Auer rods. Intracellular LZ content assayed by a post-embedding immunogold method, showed high granular LZ density (in the range of that found in M4 and M5) in M3 cells and very low granular LZ content in M3-V. This study adds new objective parameters for the diagnosis of these two types of APL and provides new information on their LZ pattern of expression. PMID- 9858225 TI - Systemic mastocytosis associated with acute myeloid leukaemia: report of two cases and detection of the c-kit mutation Asp-816 to Val. AB - A subset of patients with systemic mastocytosis (SM) develop acute myeloid leukaemia (AML). However, little is known about the biology of such leukaemias and their relationship to the mast cell (MC) lineage. We report on two female patients who suffered from SM and AML. According to FAB criteria, the leukaemias were classified as AML-M4 (patient 1) and AML-MO (patient 2). The coexistence of the two distinct neoplasms (AML and SM) was demonstrable by immunostaining of serial bone marrow (BM) sections with monoclonal antibodies (mAb). In particular, the MC infiltrates were found to react with mAb against MC-tryptase and MC growth factor receptor c-kit (CD117), but not with mAb to CD15 or CD34. In contrast, the AML blasts were immunoreactive for CD15 (patient 1) or CD34 (patient 2), but did not express tryptase. The c-kit point mutation Asp-->Val at codon 816, considered to play a role in the transformation of MC progenitors, was detected in patient 1 in a BM cell fraction containing 4% MC. However, no c-kit mutation was found in pure AML blasts (<1% MC). These findings argue against an evolution of the AML clone from neoplastic MC or MC-committed progenitors. PMID- 9858226 TI - Near-haploid common acute lymphoblastic leukaemia of childhood with a second hyperdiploid line: a DNA ploidy and fluorescence in-situ hybridization study. AB - Near-haploidy is a rare cytogenetic finding in childhood acute lymphoblastic leukaemia (ALL) and is associated with a poor prognosis. A second hyperdiploid line, occurring presumably by endoreduplication of the near-haploid stemline, is often observed. We present a case of common ALL in relapse characterized morphologically by a dual population of small and large lymphoblasts. Cytogenetic analysis supplemented with fluorescence in-situ hybridization (FISH) studies localized near-haploidy and hyperdiploidy to the small and large blast population respectively. DNA ploidy determination confirmed two abnormal clones with near haploidy as the predominant one. A novel t(9;12)(q11;q13) was present in the near haploid clone and was duplicated in the hyperdiploid clone. This finding identified cells bearing near-haploidy to be the clonogenic population following malignant transformation and confirmed endoreduplication as the mechanism for the presence of associated hyperdiploidy. PMID- 9858227 TI - Longitudinal observation and outcome of nonfamilial childhood haemophagocytic syndrome receiving etoposide-containing regimens. AB - The long-term outcome of 22 children treated with etoposide-containing regimens for haemophagocytic syndrome (HS) were longitudinally studied; none of them had a family history of the disease. All patients received etoposide-containing (150 mg/m2/d) regimens, combined, in 16 cases, with intravenous immunoglobulin (IVIG) and prednisolone. Complete remission (CR) was achieved in 12 patients, partial remission in seven, and early mortality occurred in three. Of the 12 CR patients, only four remain alive and disease-free, with a median follow-up of 47.4 months; one CR patient died due to infection and the remaining seven had relapsed diseases. Three patients with a partial response or with relapsed disease progressed to T-cell lymphoma, characterized, in the two cases tested, by clonal chromosomal abnormalities. Epstein-Barr virus (EBV) infection was implicated in disease pathogenesis in 15/22 patients. The overall survival was 45.5%, 40.9% and 40.9% at 1, 3 and 5 years, respectively, and disease-free survival for CR patients at these same times was 45.5%, 36.4% and 36.4%. The etoposide-containing regimen would appear to be an effective initial therapeutic option for childhood HS. However, in view of the frequency of partial remissions and relapsed disease, a more intensive chemotherapy or bone marrow transplantation should be applied. The progression to EBV-containing T-cell lymphoma in three patients is consistent with the previous observation that EBV-associated HS is a potentially malignant disease. PMID- 9858228 TI - The in-vitro generation of dendritic cells from blast cells in acute leukaemia. AB - Dendritic cells (DC) are potent antigen-presenting cells responsible for the initiation of primary antigen-specific immune responses. In chronic myeloid leukaemia DC have been generated from Ph+ cells and these Ph+ DC are capable of stimulating cytolytic T-cell responses against the parent leukaemia cells. The prevalence of this phenomenon in acute leukaemia (AL) is unknown and we have therefore studied a variety of acute leukaemias to determine their potential for DC development. Peripheral blood mononuclear cells (PBMC) from 21 cases of AL were cultured in GM-CSF + TNF alpha. Of these cases, 15 were viable in culture and cells with typical DC morphology were observed in 12 of these 15 cases. DC growing in culture expressed either CDla and/or CD83 and were HLA-DR+ CD40+ CD80+ CD86+ typical of mature DC. In 9/12 cases the cultured cells possessed potent antigen-presenting capacity as measured in the allo-MLR. The malignant origin of the cultured DC was confirmed by FISH analysis in two cases (one 5q- and one Ph+ AL) and by persistent aberrant expression of CD19 in two cases of biphenotypic leukaemia. Functional DC may be derived from AL blasts in a significant number of patients and such DC may be capable of inducing leukaemia-specific immune responses with potential for clinically beneficial effects. PMID- 9858229 TI - No treatment for low-risk thrombocythaemia: results from a prospective study. AB - Essential thrombocythaemia (ET) is a chronic myeloproliferative disorder characterized by the occurrence of thromboembolic episodes, particularly in patients aged > 60 years or with a previous history of thrombosis, and/or by haemorrhages in patients with an exceedingly high platelet count. In these subgroups of patients the use of cytoreductive therapy is beneficial in terms of risk/benefit ratio. Only limited anecdotal data are available on the thrombotic or haemorrhagic risk and survival in young asymptomatic ET patients with a platelet count < 1500 x 10(9)/l. Therefore the optimal management of these patients is unknown. To assess the incidence of thrombosis and haemorrhages in this group of patients we carried out a prospective observational study in a cohort of 65 patients with ET, aged < 60 years, with no history of thrombosis or haemorrhage and platelet count < 1500 x 10(9)/l, and in 65 age- and sex-matched controls. Patients were not treated with cytoreductive therapy until the occurrence of thrombosis or haemorrhage. Arterial or venous thrombotic events were objectively documented both in cases and in controls. The median follow-up was 4.1 years, with an incidence of thrombosis in patients and controls of 1.91 and 1.50 cases/100 patient-years, respectively. The age- and sex-adjusted risk rate ratio was 1.43 (95% CI 0.37-5.4). Only three minor haemorrhagic episodes occurred in patients, with an incidence of 1.12 cases/100 patient-years. Pregnancy and surgery were not associated with thrombosis in these patients. We conclude that the thrombotic risk in young ET patients, with no thrombotic history and a platelet count < 1500 x 10(9)/l, is not increased compared to the normal population and that a conservative therapeutic approach should therefore be considered in these patients. PMID- 9858230 TI - Plasma thrombopoietin (TPO) levels and expression of TPO receptor on platelets in patients with myelodysplastic syndromes. AB - Data on endogenous thrombopoietin (TPO) levels and their regulation in myelodysplastic syndromes (MDS) are sparse. We examined the plasma TPO level of 85 MDS patients by a sensitive enzyme immunoassay and the platelet expression of TPO receptor (TPO-R) protein, which metabolizes endogenous TPO, in 19 MDS patients with an equilibrium binding assay using 125I-TPO. The MDS patients had higher plasma TPO levels (7.0 +/- 9.3 fmol/ml) than 52 normal subjects (P < 0.0001). Refractory anaemia (RA) patients (n = 39) had higher plasma TPO levels than patients (n = 28) with RA with excess blasts (RAEB) or RAEB in transformation (RAEB-t) (P = 0.0002), irrespective of similar platelet counts in these groups. The plasma TPO level correlated inversely with the platelet count in RA patients (P = 0.0027) but not in RAEB and RAEB-t patients (P = 0.7865). These data suggest that the physiological pathway for TPO production and metabolism is conserved, at least partially, in RA, but deranged in RAEB/RAEB-t. The number of TPO-R per platelet was significantly smaller in 19 MDS patients (17.5 +/- 13.3) than in normals (P = 0.0014), but similar between RA patients and patients with RAEB and RAEB-t. Further, the bone marrow megakaryocyte count, determined in 31 MDS patients, was quite similar between RA patients and patients with RAEB or RAEB-t. Thus, in addition to thrombocytopenia, a reduced platelet TPO-R number may contribute to elevated plasma TPO levels in MDS, and a regulatory pathway for circulating TPO other than platelet TPO-R and marrow megakaryocytes, such as blasts expressing TPO-R, may operate in RAEB/RAEB-t. PMID- 9858231 TI - Poor response rate to a continuous schedule of Amifostine therapy for 'low/intermediate risk' myelodysplastic patients. AB - Ineffective haemopoiesis leading to cytopenia presents the major clinical management problem for patients with myelodysplasia (MDS). Preliminary studies have demonstrated that the synthetic aminothiol Amifostine stimulates multilineage haemopoiesis both in vitro and in vivo in patients with MDS. We have treated 12 patients with an uninterrupted 8-week schedule of thrice-weekly intravenous Amifostine with a starting dose of 300 mg/m2 escalating to 450 mg/m2 in non-responders. No patients satisfied response criteria on study but two patients showed minor responses. We conclude that therapeutic response to Amifostine in MDS may be schedule dependent. PMID- 9858232 TI - Reactivation of Behcet's disease in the course of multicentric HHV8-positive Castleman's disease: long-term complete remission by a combined chemo/radiation and interferon-alpha therapy regimen. AB - We used a new combined chemo- (COP/ABVD), radiation and interferon-alpha (10 x 10(6) IU s.c. 3x per week/12 months) therapy regimen to treat severe multicentric Castleman's disease (CD) complicated by relapsing Behcet's disease (BD) manifestations. More than 16 years after diagnosis of CD the patient remains in very good clinical condition, with remission of all CD and BD manifestations 13 months after discontinuation of the interferon-alpha treatment. In addition, our clinicopathological, immunohistological and virological data suggest a pathogenetic link between CD and BD via activation of pre-existing BD-specific plasma cells due to CD-related HHV8-induced overexpression of interleukin-6. PMID- 9858233 TI - A novel BCR-ABL fusion gene (e2/1a) in a patient with Philadelphia-positive chronic myelogenous leukaemia and an aggressive clinical course. AB - A novel variant BCR-ABL mRNA transcript was detected by reverse transcription polymerase chain reaction (RT-PCR) in a patient with Philadelphia positive (Ph+ve) chronic myelogenous leukaemia (CML) who had an aggressive clinical course. Sequence analysis of the amplified cDNA fragment revealed an in-frame fusion with part of BCR exon e2 joined to part of ABL exon 1a. PCR of genomic DNA from this patient demonstrated that this unusual chimaeric mRNA resulted from breakpoints that fell within each of these exons. This is the first report of breakpoints occurring within both ABL and BCR exons and the first fusion to involve ABL exon 1a. PMID- 9858234 TI - Cerebral candidiasis in a child 1 year after leukaemia. AB - We describe an unusual case of a late presentation of a fungal brain abscess in a non-neutropenic child 1 year after completing chemotherapy for M5 acute myeloid leukaemia (AML). Biopsy of the mass identified candidal hyphae and the patient was treated with 5 mg/kg of liposomal amphotericin B for 6 weeks. The lesion resolved completely and the child remains well 2 years later. Invasive fungal infection should be included in the differential diagnosis of unexplained symptoms in patients who have previously received intensive chemotherapy. PMID- 9858235 TI - Mast cell recruitment after subcutaneous injection of RANTES in the sole of the rat paw. AB - The effect of hrRANTES was studied after the injection in the sole of the rat paw, an area particularly rich in mast cells. Subcutaneous injections of RANTES 50 ng/10 microl produced an erythematous reaction which was inhibited by anti RANTES antibody 50 microg/rat injected in the tail vein 30 min before hrRANTES 50 ng/10 microl was injected. In another set of experiments the animals were injected subcutaneously in the sole of the paw with PBS 10 microl (control), LPS (100 ng/10 microl) hrRANTES 50 ng/10 microl or anti-RANTES 50 microl/rat injected in the tail vein 30 min before hrRANTES 50 ng/10 microl was injected. The biopsies were analysed after 4 h and counted in an optic field. hrRANTES produced a strong recruitment of mast cells selectively coloured with 0.1% toluidine blue and inhibited by anti-RANTES antibody. In addition to the optical and electron microscope study, in some of the excised tissue Northern blot analysis for histidine decarboxylase (HDC) mRNA was performed to estimate the amount of histamine generation in the tissue of the injection sites. We found that subcutaneous injections of hrRANTES 50 ng/10 microl in the sole of the rat paw produced an accumulation of a great number of mast cells compared to PBS 10 microl (negative control) or LPS 100 ng/10 microl (positive control) after 4 h. The hrRANTES effect was inhibited by anti-RANTES antibody injected in the tail vein 30 min before hrRANTES exposure. Moreover, hrRANTES increased HDC mRNA and histamine generation. PMID- 9858236 TI - Effect of the interaction between fibronectin and VLA-4 on the proliferation of human B cells, especially a novel human B-cell line, OPM-3. AB - Very late antigen (VLA)-4 integrin has been suggested to play an important role in haemopoiesis. However, little is known concerning the roles of the fibronectin (FN)/VLA-4 interaction in the proliferation of human B cells. In this study we investigated the effect of immobilized FN on the proliferation of various B-cell lines, including a newly-established B-cell line, OPM-3, and human tonsillar B cells, that primarily express VLA-4 but not VLA-5. Immobilized FN significantly promoted the proliferation of OPM-3 cells and normal B cells via VLA-4. The cross linking of beta1 integrins of OPM-3 cells resulted in the phosphorylation of the focal adhesion kinase (FAK) associated 90 kD protein, an increase in FAK associated kinase activity, and the phosphorylation of Raf-1. Furthermore, the MEK1 inhibitor, PD98059, inhibited the FN-promoted proliferation of OPM-3 cells. These results demonstrate that the FN/VLA-4 interaction transmits the growth signal(s) which may be mediated by Ras pathway in OPM-3 cells, and suggest that OPM-3 cells may be of great value in studying the roles of the FN/VLA-4 interaction in human B-cell growth. PMID- 9858237 TI - The haemochromatosis mutations do not modify the clinical picture of thalassaemia major in patients regularly transfused and chelated. AB - Iron overload is the main cause of morbidity and mortality in patients with thalassaemia major. In order to establish if the presence of the mutations recently described in the haemochromatosis gene affects the severity of iron overload in thalassaemia patients, we compared the prevalence of mutations C282Y and H63D in 216 young adults regularly transfused and chelated in North-Eastern Italy with the frequency found in a group of blood donors from the same area. For each patient, mean serum ferritin over the last 3 years, liver iron concentration, and the presence of diabetes, hypogonadism and heart disease, were considered. The frequency of the C282Y allele was 1.9% in patients with thalassaemia major and 2.3% in blood donors (P=ns). The frequency of the H63D allele was 16.2% in patients with thalassaemia major and 15.3% in blood donors (P=ns). When age, liver iron concentration and mean yearly serum ferritin levels were compared in patients with and without mutations C282Y and H63D, no significant differences were found. Also, the prevalence of iron-induced complications was not significantly different between patients carrying or not carrying the mutations. The presence of the HH mutations does not seem to influence the degree of iron overload and its consequences in regularly transfused and chelated patients with thalassaemia major. PMID- 9858238 TI - Iron status in pregnant women: which measurements are valid? AB - Anaemia in pregnancy in developing countries continues to be a public health problem of significant proportion. At least 50% of the anaemia has been blamed on iron deficiency. In populations where chronic inflammation and iron deficiency anaemia coexist, the criteria to accurately define iron status are not always clear. Similarly, in pregnancy, with marked physiological changes, cut-off points for biochemical parameters need to be re-examined. In this study we examined the diagnostic accuracy of iron parameters including mean cellular volume (MCV), serum iron, transferrin, total iron binding capacity (TIBC) and its saturation, zinc protoporphyrin (ZPP), ferritin and serum transferrin receptor (TfR) for the assessment of iron status in a population of anaemic pregnant women in Malawi. Stained bone marrow aspirates were used as the standard for comparison. Results show that for the purpose of screening, serum ferritin is the best single indicator of storage iron provided a cut-off point of 30 microg/l is used. A number of other commonly used parameters of iron status were shown to have limited diagnostic accuracy. Logistic regression was used to obtain mathematical models for the prediction of bone marrow iron status using a combination of available parameters. PMID- 9858239 TI - Effects of four species of interferon-alpha on cultured erythroid progenitors from congenital dyserythropoietic anaemia type I. AB - The in vitro effects of interferon (IFN)-alpha on erythroid progenitor cells derived from the peripheral blood of five patients with congenital dyserythropoietic anaemia (CDA) type I and seven healthy adults were studied. Ficol-hypaque-separated mononuclear cells were cultured for 14 d in StemGEM-1d medium with 0, 1, 10 and 100 U/ml of recombinant IFN-alpha2a, recombinant IFN alpha2b, the genetically-engineered hybrid molecule IFN-alpha(1-8) or the laboratory-designed molecule IFN-alpha(consensus). Erythroid bursts and colonies were counted, picked and processed for electron microscopy. In the experiments employing IFN-alpha2a there were no differences in the numbers of erythroid bursts or colonies between four patients with CDA type I and seven healthy adults. All five patients with CDA type I showed the 'Swiss-cheese' ultrastructural abnormality of the heterochromatin in a proportion of the erythroblasts when the progenitor cells were cultured in the absence of added IFN alpha. A statistically significant reduction in the proportion of erythroblasts showing the 'Swiss-cheese' defect was seen when the erythroid progenitors were cultured in the presence of 0.01-0.5 U/ml IFN-alpha2a (five patients), or 0.1 U/ml of IFN-alpha2b (two patients). In contrast, no reduction was seen in cultures containing 0.1-100 U/ml of either IFN-alpha(1-8) or IFN-alpha(consensus) or 20 microM hydroxyurea (two patients). The partial correction of the 'Swiss cheese' abnormality by low concentrations of IFN-alpha2a in vitro provides an experimental model with which the mechanisms underlying the haematological response that occurs after the in vivo administration of this species of IFN alpha may be investigated. PMID- 9858240 TI - Congenital dyserythropoiesis with intererythroblastic chromatin bridges and ultrastructurally-normal erythroblast heterochromatin: a new disorder. AB - Two non-anaemic subjects, a father and daughter, with a new form of congenital dyserythropoiesis are reported. The features of their disorder are: (1) an abnormal blood film with basophilic stippling of red cells and oval macrocytes, (2) various dysplastic changes in the erythroblasts, including internuclear chromatin bridges, (3) ultrastructurally-normal erythroblast heterochromatin, (4) normal serum thymidine kinase activity, and (5) a probable autosomal dominant inheritance. The last three features distinguish this disorder from CDA type I. PMID- 9858241 TI - Activation of the human delta-globin gene promoter in primary adult erythroid cells. AB - Restoration of the CCAAT box or insertion of an erythroid Kruppel-like factor (EKLF) binding site in the delta promoter activates its expression in several erythroid cell lines. We extended these studies using a novel primary human adult erythroid cell (hAEC) system to investigate these effects at the late erythroblast stage. Restoration of the CCAAT box at -70 bp, or insertion of an EKLF binding site at -85 bp or -95 bp in the promoter significantly increased delta globin gene expression in hAEC. Our results demonstrate that the altered CCAAT box (CCAAC) and the lack of an EKLF binding site in delta-globin contribute to its low level of expression in the hAEC model as well. PMID- 9858242 TI - R411C mutation of the ALAS2 gene encodes a pyridoxine-responsive enzyme with low activity. AB - A R411C missense mutation of the erythroid-specific delta-aminolaevulinate synthase (ALAS2) gene was identified in a pedigree with X-linked pyridoxine responsive sideroblastic anaemia (XLSA). The normal and the mutant cDNAs were expressed in E. coli, and the enzyme protein was purified. ALAS activity of the mutant enzyme was 12% and 25%, when incubated in the absence and the presence of pyridoxal 5'-phosphate, respectively, compared with that of the wild-type enzyme. These findings suggest that the R411C mutation accounts for low ALAS activity and a partial pyridoxine-responsiveness of the disease in the patient. PMID- 9858243 TI - Prevalence of the C282Y and H63D mutations in the HFE gene in patients with hereditary haemochromatosis and in control subjects from Northern Germany. AB - Mutation analysis was performed for two HFE mutations (C282Y, H63D) in unrelated patients with hereditary haemochromatosis (n = 92), family members of patients (n = 34), and unrelated controls (n = 157) from Northern Germany, 87/92 patients (94.6%) revealed the C282Y mutation in homozygous form, five were heterozygous. No H63D mutation was found in 174 chromosomes of patients homozygous for C282Y, whereas four of the heterozygote patients also carried the H63D mutation. Among the control group, 9.6% were heterozygotes for C282Y. 2/157 subjects were homozygous, 37/157 were heterozygous for the H63D mutation, but showed no signs of iron overload. PMID- 9858244 TI - The incidence and significance of fevers during treatment with antithymocyte globulin for aplastic anaemia. AB - Antithymocyte globulin (ATG) is a foreign protein used widely to treat aplastic anaemia (AA). Febrile reactions occurring during its administration may be impossible to distinguish clinically from fever due to sepsis, and are therefore routinely treated with intravenous antibiotics after collection of blood cultures. A statistically highly significant difference was found in positive blood cultures between 39 AA patients who developed fever during ATG therapy, and 38 febrile neutropenic patients with acute leukaemia. suggesting that most fevers developing during ATG treatment are not due to infection. It may therefore be reasonable to consider early discontinuation of intravenous antibiotics in patients who are clinically stable and have no proven sepsis. PMID- 9858245 TI - Anti-CD36 autoantibodies in thrombotic thrombocytopenic purpura and other thrombotic disorders: identification of an 85 kD form of CD36 as a target antigen. AB - The presence of anti-CD36 antibodies in plasma of patients with thrombotic thrombocytopenic purpura (TTP), idiopathic thrombocytopenic purpura (ITP), and heparin-induced thrombocytopenia without/with thrombosis (HIT/HITT) has been examined by immunoblots, and a monoclonal antibody capture assay, the platelet associated IgG characterization assay (PAICA). Results with PAICA showed that 73% (8/11) of patients with TTP were positive, and 71% (10/14) by immunoblots. With ITP, 20% (6/30) were positive by PAICA and 19% (3/16) by immunoblots; HIT, 30% (3/10) were positive by PAICA and 60% (6/10) by immunoblot; HITT, 50% (2/4) by PAICA and 100% (4/4) by immunoblot. Purification of CD36 by fast protein liquid chromatography (FPLC) from Triton X-100 extracts of normal platelet membranes resulted in the isolation of two different forms: the classic 88 kD form, and a second, lighter 85 kD form. Our data indicated that the patients' plasma autoantibodies reacted strongly with the 85 kD form. Conventional monoclonal and polyclonal antisera produced to the 88 kD form reacted strongly with the 88 kD form but weakly with the 85 kD form. These results confirm the possible importance of anti-CD36 antibodies in the pathophysiology of TTP and other thrombocytopenias and demonstrate the presence of a previously unrecognized target antigen for these antibodies. PMID- 9858246 TI - The P2Y1 receptor, necessary but not sufficient to support full ADP-induced platelet aggregation, is not the target of the drug clopidogrel. AB - Recently we showed that the P2Y1 receptor coupled to calcium mobilization is necessary to initiate ADP-induced human platelet aggregation. Since the thienopyridine compound clopidogrel specifically inhibits ADP-induced platelet aggregation, it was of interest to determine whether the P2Y1 receptor was the target of this drug. Therefore we studied the effects of clopidogrel and of the two specific P2Y1 antagonists A2P5P and A3P5P on ADP-induced platelet events in rats. Although clopidogrel treatment (50 mg/kg) greatly reduced platelet aggregation in response to ADP as compared to untreated platelets, some residual aggregation was still detectable. In contrast, A2P5P and A3P5P totally abolished ADP-induced shape change and aggregation in platelets from both control and clopidogrel-treated rats. A2P5P and A3P5P (100 microM) totally inhibited the [Ca2+]i rise induced by ADP (0.1 microM) in control and clopidogrel-treated platelets, whereas clopidogrel treatment had no effect. Conversely, the inhibition of adenylyl cyclase induced by ADP (5 microM) was completely blocked by clopidogrel but not modified by A2P5P or A3P5P (100 microM). A3P5P (1 mM) reduced the number of [33P]2MeSADP binding sites on control rat platelets from 907 +/- 50 to 611 +/- 25 per platelet. After clopidogrel treatment, binding of [33P]2MeSADP decreased to 505 +/- 68 sites per platelet and further decreased to 55 +/- 12 sites in the presence of A3P5P (1 mM). In summary, these results demonstrate that the platelet P2Y1 receptor responsible for the initiation of aggregation in response to ADP is not the target of clopidogrel. Platelets may express another, as yet unidentified, P2Y receptor, specifically coupled to the inhibition of adenylyl cyclase and necessary to induce full platelet aggregation, which could be the target of this drug. PMID- 9858247 TI - Plasminogen Kanagawa-I, a novel missense mutation, is caused by the amino acid substitution G732R. AB - A new dysplasminogen, plasminogen Kanagawa-I, was identified in a healthy male with no previous thrombotic episodes. His plasma plasminogen (PLG) activity was 51.4% of that of normal pooled plasma (reference interval 70-130%) and the antigen level was 94.2% of that of normal pooled plasma (reference interval 80 150%). Nucleotide sequencing revealed a heterozygous G to A transition in exon 18, which resulted in an amino acid substitution of G732R. Both the proband's father and paternal grandfather were heterozygous for this mutation. Interestingly, the grandfather was found to be a compound heterozygote for plasminogen Kanagawa-I and Tochigi (A601T), so that his plasminogen activity and antigen level was 7.7% and 87.2% of that of normal pooled plasma, respectively. However, he has never been affected by significant thrombosis. PMID- 9858249 TI - Identification of new type 2B von Willebrand disease mutations: Arg543Gln, Arg545Pro and Arg578Leu. AB - We report the identification in five patients (three families) affected with type 2B von Willebrand disease (VWD) of three heterozygous nucleotide substitutions at the codon for arginine 543, 545 and 578 of the mature von Willebrand factor (VWF) subunit resulting in a glutamine, proline and leucine substitution, respectively. These mutations are located in the A1 loop where prevalent type 2B mutations (Arg543Trp, Arg545Cys and Arg578Gln) have been already identified at the same positions. By in vitro mutagenesis of full-length cDNA of VWF and transient expression in Cos-7 cells, we have shown that the six corresponding mutated recombinant VWFs (Gln543, Trp543, Cys545, Pro545, Leu578 and Gln578 rVWF) exhibited quantitatively normal expression and normal multimeric pattern but increased ristocetin- and botrocetin-induced binding to platelets as compared with that for wild-type rVWF. The two mutations at position 545 induced the greatest reactivity for GPIb of corresponding rVWFs as compared to the two mutations at positions 543 and 578. PMID- 9858248 TI - Additional genetic risk factors for venous thromboembolism in carriers of the factor V Leiden mutation. AB - Only a minority of subjects with factor V (FV) Leiden mutation develop venous thromboembolism (VTE), suggesting that additional genetic risk factors may be present in symptomatic carriers. We screened 157 unrelated carriers of the FV Leiden mutation with a first episode of VTE and 291 unrelated asymptomatic FV carriers for the presence of two frequent mutations, i.e. G20210A of the prothrombin gene and C677T of the methylenetetrahydrofolate reductase gene. Carriers with other inherited or acquired thrombophilia-associated abnormalities were excluded from analysis. Heterozygotes for the G20210A mutation were more prevalent among symptomatic carriers than in asymptomatic carriers (10.8% v 2.7%, P<0.0001); homozygotes for the C677T mutation were also more prevalent in symptomatic carriers (21.6% v 14.4%, P = 0.05). Factor V Leiden carriers who had had a VTE episode during oral contraceptive intake were more frequently carriers of the G20210A mutation (14.3%, P = 0.03). These results further support the idea that VTE in carriers of FV Leiden results from interaction with additional genetic or circumstantial risk factors, and that an accurate search for such factors is required to identify carriers at risk. PMID- 9858250 TI - Two novel mutations (Pro864His, Val867Glu) causing type 2A von Willebrand disease and affecting a single restriction site in exon 28. AB - We detected two transversions in two unrelated Italian patients with type 2A von Willebrand disease (VWD): a C to A at nucleotide 8821 and a T to A at nucleotide 8830, resulting in the missense mutations Pro864His and Val867Glu respectively. Both mutations were in the heterozygous form and abolished the BstXI restriction site in exon 28 of the VWF gene. In both mutations plasma VWF multimer pattern improved by antiproteases. Moreover, DDAVP normalized plasma VWF multimers in the Pro864His patient, especially when protease inhibitors were present. These new mutations appear to be of the 2A VWD subtype due to the increased susceptibility to proteases. PMID- 9858251 TI - Factor V Arg306-->Thr (factor V Cambridge) and factor V Arg306-->Gly mutations in venous thrombotic disease. AB - We investigated the prevalence of two reported mutations of the factor V gene (factor V Arg306-->Thr, or factor V Cambridge, and factor V Arg306-->Gly) in 104 relatively young patients with verified venous thrombosis and in 208 age-, sex- and race-matched controls, in order to establish whether the two mutations are associated with increased predisposition for venous thrombosis. PCR amplification followed by BstNI and MspI digestion was employed to determine the genotypes, and each mutation was confirmed by DNA sequencing. Among the controls, one individual was found to be heterozygous for the factor V Arg306-->Thr mutation and one heterozygous for the factor VArg306-->Gly mutation; none of the patients carried either mutation. Our findings do not support factor V Cambridge and factor V Arg306-->Gly as risk factors for venous thrombosis. PMID- 9858252 TI - Acute liver failure induced by alcohol and paracetamol in an HCV-infected haemophiliac. AB - Paracetamol (acetaminophen) is frequently used by haemophiliacs in the management of acute or chronic pain, primarily due to its lack of adverse effects when taken at therapeutic doses. We describe acute hepatic and renal toxicity of paracetamol in a 36-year-old patient with haemophilia B, chronic hepatitis C and chronic alcohol abuse. Moderate doses of paracetamol (6 g/d for 4 d), taken with therapeutic intent, resulted in life-threatening organ dysfunction which gradually recovered with full supportive care. PMID- 9858253 TI - Complete splenic embolization in the treatment of immune thrombocytopenic purpura. PMID- 9858254 TI - Merlin, the neurofibromatosis type 2 gene product, and beta1 integrin associate in isolated and differentiating Schwann cells. AB - Neurofibromatosis type 2, a disease characterized by the formation of multiple nervous system tumors, especially schwannomas, is caused by mutation in the gene encoding merlin/schwannomin. The molecular mechanism by which merlin functions as a tumor suppressor is unknown, but is hypothesized to involve plasma membrane and cytoskeleton interaction. Several merlin antibodies were used to study merlin expression, localization, and protein association in primary cultures of rat sensory neurons, Schwann cells (SCs), and SCs grown with neurons (SC/N cultures) before and during differentiation into myelinating cells. Western blot analysis revealed that neurons predominantly expressed a 68-kD protein, but SCs expressed two additional 88- and 120-kD related proteins. Extensive immunological characterization demonstrated that the 88-kD protein shared three domains with the 68-kD merlin protein. Western blot analysis of soluble and insoluble culture fractions demonstrated that the majority of merlin and related proteins were soluble in isolated SCs and undifferentiated SC/N cultures, but became insoluble in myelinating SC/N cultures. Double immunofluorescence staining suggested that merlin translocated from the perinuclear cytoplasm in undifferentiated SCs to the subplasmalemma in differentiating SCs and partially colocalized with beta1 integrin. Finally, beta1 integrin antibody coimmunoprecipitated 68-kD merlin from isolated SC and undifferentiated SC/N cultures, but predominantly the 88-kD protein from differentiating SC/N cultures. Together, these results provide evidence that merlin interacts with beta1 integrin and that merlin localization changes from a cytosolic to cytoskeletal compartment during SC differentiation. PMID- 9858255 TI - Identification of reggie-1 and reggie-2 as plasmamembrane-associated proteins which cocluster with activated GPI-anchored cell adhesion molecules in non caveolar micropatches in neurons. AB - Neurons are believed to possess plasmalemmal microdomains and proteins analogous to the caveolae and caveolin of nonneuronal cells. Caveolae are plasmalemmal invaginations where activated glycosyl-phosphatidylinositol (GPI)-anchored proteins preferentially assemble and where transmembrane signaling may occur. Molecular cloning of rat reggie-1 and -2 (80% identical to goldfish reggie proteins) shows that reggie-2 is practically identical to mouse flotillin-1. Flotillin-1 and epidermal surface antigen (ESA) (flotillin-2) are suggested to represent possible membrane proteins in caveolae. Rat reggie-1 is 99% homologous to ESA in overlapping sequences but has a 49-amino-acid N-terminus not present in ESA. Antibodies (ABs) which recognize reggie-1 or -2 reveal that both proteins cluster at the plasmamembrane and occur in micropatches in neurons [dorsal root ganglia (DRGs), retinal ganglion, and PC-12 cells] and in nonneuronal cells. In neurons, reggie micropatches occur along the axon and in lamellipodia and filopodia of growth cones, but they do not occur in caveolae. By quantitative electronmicroscopic analysis we demonstrate the absence of caveolae in (anti caveolin negative) neurons and show anti-reggie-1 immunogold-labeled clusters at the plasmamembrane of DRGs. When ABs against the GPI-anchored cell adhesion molecules (CAMs) F3 and Thy-1 are applied to live DRGs, the GPI-linked CAMs sequester into micropatches. Double immunofluorescence shows a colocalization of the CAMs with micropatches of anti-reggie antibodies. Thus, reggie-1 and reggie-2 identify sites where activated GPI-linked CAMs preferentially accumulate and which may represent noncaveolar micropatches (domains). PMID- 9858256 TI - Rac1-dependent actin filament organization in growth cones is necessary for beta1 integrin-mediated advance but not for growth on poly-D-lysine. AB - The activity of filopodia and lamellipodia determines the advance, motility, adhesion, and sensory capacity of neuronal growth cones. The shape and dynamics of these highly motile structures originate from the continuous reorganization of the actin cytoskeleton in response to extracellular signals. The small GTPases, Rac1, Rho, and CDC42, regulate the organization of actin filament structures in nonneuronal cells; yet, their role in growth cone motility and neurite outgrowth is poorly understood. We investigated in vitro the function of Rac1 in neurite outgrowth and differentiation by introducing purified recombinant mutants of Rac1 into primary chick embryo motor neurons via trituration. Endogenous Rac1 was expressed in growth cone bodies as well as in the tips and shafts of filopodia, where it often colocalized with actin filament structures. The introduction of constitutively active Rac1 resulted in an increase in rhodamine-phalloidin staining, presumably from an accumulation of actin filaments in growth cones, while dominant negative Rac1 caused a decrease in rhodamine-phalloidin staining. Nevertheless, both Rac1 mutants retarded growth cone advance, and hence attenuated neurite outgrowth and inhibited differentiation of neurites into axons and dendrites on laminin and fibronectin. In contrast, on poly-D-lysine, neither Rac1 mutant affected growth cone advance, neurite outgrowth, or neurite differentiation despite inducing similar changes in the amount of rhodamine phalloidin staining in growth cones. Our data demonstrate that Rac1 regulates actin filament organization in neuronal growth cones and is pivotal for beta1 integrin-mediated growth cone advance, but not for growth on poly-D-lysine. PMID- 9858257 TI - Response of retinal ganglion cell axons to striped linear gradients of repellent guidance molecules. AB - Although molecular gradients have long been postulated to play a role in the development of topographic projections in the nervous system, relatively little is known about how axons evaluate gradients. Do growth cones respond to concentration or to slope? Do they react suddenly or gradually? Is there adaptation? In the developing retinotectal system, temporal retinal ganglion cell axons have previously been shown to avoid repellent cell-surface activities distributed in gradients across the optic tectum. We confronted temporal retinal axons with precisely formed striped linear gradients of repellent tectal membranes and of two candidate repellent molecules, ephrin-A2 and -A5. Axons entered gradient stripes independently of their slope and extended unhindered in the uphill direction until they suddenly avoided an apparent threshold concentration of repellent material that was independent of slope. This critical concentration was similar in both linear and nonlinear gradients, and hence independent of gradient shape. When gradients of identical slope were formed on different basal levels of repellent material, axons grew uphill for a fixed increment of concentration, possibly measured from the lowest point of the gradient, rather than up to a fixed absolute concentration. The speed of growth cones was not affected by repellent unstriped gradients below the critical concentration level. Similar results were found with membranes from cell lines stably transfected with either ephrin-A5 or ephrin-A2, two previously identified growth cone repellent cell-surface proteins. These data suggest that growth cones or axons can integrate guidance information over large distances, probably by a combined memory and adaptation mechanism. PMID- 9858258 TI - TGF-beta regulates the survival of ciliary ganglionic neurons synergistically with ciliary neurotrophic factor and neurotrophins. AB - We investigated putative roles of transforming growth factor (TGF)-beta expressed in peripheral ganglia in the regulation of neuronal cell survival during the period of ontogenetic neuron death (OD). The chick ciliary ganglion (CG), where OD occurs between embryonic days (E) 6 and 10, was employed as a model system. We show that CG neurons (E8) are immunoreactive (ir) for TGF-beta2 and -beta3 as well as the TGF-beta receptor TbetaR-II, but are not ir for TGF-beta1. Ciliary neurotrophic factor (CNTF) and fibroblast growth factor (FGF)-2, established neurotrophic molecules for CG neurons, up-regulate TGF-beta3 mRNA and TGF-beta biological activity in cultures of E8 CG neurons. None of the TGF-beta isoforms- beta1, beta2, or beta3--has a trophic, survival-promoting effect on cultured CG neurons. However, all isoforms enhance CG neuron survival mediated by CNTF or FGF 2, significantly and over a wide range of concentrations. In combination with the neurotrophins (NT) nerve growth factor (NGF) and NT-3, which are not neurotrophic for CG neurons, TGF-beta significantly promotes CG neuron survival. However, TGF beta does not act synergistically with the neuropoietic cytokines oncostatin M, leukemia inhibiting factor, or interleukin-6. Immunoneutralization of endogenous TGF-beta released from CG neurons using an antibody to TGF-beta1/-beta2/-beta3 significantly reduces the potency of CNTF or FGF-2 to promote CG neuron survival. The blocking effect of the anti-pan-TGF-beta antibody could be rescued by adding exogenous TGF-beta. Together, these data suggest that para-/autocrine TGF-beta signaling has an important effect on the regulation of neuron survival in a model system of peripheral neurons. PMID- 9858259 TI - Fibroblast growth factor-2 stimulates cell proliferation and decreases sexually dimorphic cell death in an avian song control nucleus. AB - The neural system controlling song in birds has proven a useful model for investigating how neuronal growth and survival are regulated by sexual differentiation. The present study focused on one song control area, the robust nucleus of the archistriatum (RA), and explored how sex differences in the proliferation of putative glia cells in this region influence sexually dimorphic cell survival. In zebra finches (Poephila guttata), RA neuron death is much greater in young females than in males, resulting in marked sex differences in RA neuron number. An earlier study indicated that just prior to this sexually dimorphic neuron death the proliferation of putative glia cells within the RA is significantly lower in females than in males and remains so throughout the peak of neuron death. This suggests that sex differences in glia (or glia-derived molecules) might regulate neuron survival during sexual differentiation of the RA. To determine whether increased cell proliferation within the RA favors increased cell survival, we infused the potent glia mitogen fibroblast growth factor-2 (FGF-2) into the RA unilaterally in young females. We find that FGF-2 infusions increase RA cell proliferation and concurrently decrease the incidence of degenerating RA cells, results consistent with the hypothesis that glia exert neurotrophic effects on RA neurons during sexual differentiation. PMID- 9858260 TI - Sexually dimorphic responses to neonatal basal forebrain lesions in mice: I. Behavior and neurochemistry. AB - The nucleus basalis magnocellularis (nBM) provides the primary source of cholinergic input to the cortex. Neonatal lesions of the nBM produce transient reductions in cholinergic markers, persistent abnormalities in cortical morphology, and spatial navigation impairments in adult mice. The present study examined sex differences in the effects of an electrolytic nBM lesion on postnatal day 1 (PND 1) in mice on behavior and neurochemistry in adulthood. Mice were lesioned on PND 1 and tested at 8 weeks of age on a battery of behavioral tests including passive avoidance, cued and spatial tasks in the Morris water maze, simple and delayed nonmatch to sample versions of an odor discrimination task, and locomotor activity measurements. Following behavioral testing, mice were sacrificed for either morphological assessment or neurochemical analysis of a cholinergic marker or catecholamines. There were no lesion or sex differences in acquisition or retention of passive avoidance, performance of the odor discrimination tasks, or activity levels. Control mice showed a robust sex difference in performance of the spatial water maze task. The lesion produced a slight cued but more dramatic spatial navigation deficit in the water maze which affected only the male mice. Neurochemical analyses revealed no lesion-induced changes in either choline acetyltransferase activity or levels of norepinephrine or serotonin at the time of testing. The subsequent report shows a sex difference in lesion-induced changes in cortical morphology which suggests that sexually dimorphic cholinergic influences on cortical development are responsible for the behavioral deficits seen in this study. PMID- 9858261 TI - Sexually dimorphic responses to neonatal basal forebrain lesions in mice: II. Cortical morphology. AB - Previous studies in the mouse have shown that neonatal lesions to the cholinergic basal forebrain (nBM) areas result in transient cholinergic depletion of neocortex and precipitate altered cortical morphogenesis. Lesion-induced morphological alterations in cortex persist into adulthood and are accompanied by behavioral changes, including spatial memory deficits. The current study investigated whether neonatal nBM lesions affect male and female mice differently in adulthood. Quantitative morphometry of cortical layer width was employed to assess alterations in cytoarchitecture in neonatally nBM-lesioned and littermate control mice of both sexes following behavioral testing. Our results showed significant decreases in cortical layer IV and V widths across somato/motor cortex in neonatally nBM lesioned mice of both sexes. Sexually dimorphic responses were observed in cortical layer II/III and total cortical width, limited to the area containing the "barrel cortex" representation of the whisker hairs. In lesioned females, layer II/III and total cortical width were decreased relative to female controls, and in lesioned males, layer II/III was increased relative to controls, whereas total cortical width was unchanged. In male but not female mice we observed significant correlations between decreased widths in layer IV and V and impaired performance on a spatial memory task. The current data further support a role of developing cholinergic cortical afferents in the modulation of cortical morphogenesis and cortical circuits involved in cognitive behaviors. In addition, our observations provide further evidence for sexually dimorphic development and function in cognitive centers of the rodent brain. PMID- 9858262 TI - In vivo dynamics of axon pathfinding in the Drosophilia CNS: a time-lapse study of an identified motorneuron. AB - We developed a system for time-lapse observation of identified neurons in the central nervous system (CNS) of the Drosophila embryo. Using this system, we characterize the dynamics of filopodia and axon growth of the motorneuron RP2 as it navigates anteriorly through the CNS and then laterally along the intersegmental nerve (ISN) into the periphery. We find that both axonal extension and turning occur primarily through the process of filopodial dilation. In addition, we used the GAL4-UAS system to express the fusion protein Tau-GFP in a subset of neurons, allowing us to correlate RP2's patterns of growth with a subset of axons in its environment. In particular, we show that RP2's sharp lateral turn is coincident with the nascent ISN. PMID- 9858263 TI - Time course of the development of motor behaviors in the zebrafish embryo. AB - The development and properties of locomotor behaviors in zebrafish embryos raised at 28.5 degrees C were examined. When freed from the chorion, embryonic zebrafish showed three sequential stereotyped behaviors: a transient period of alternating, coiling contractions followed by touch-evoked rapid coils, then finally, organized swimming. The three different behaviors were characterized by video microscopy. Spontaneous, alternating contractions of the trunk appeared suddenly at 17 h postfertilization (hpf), with a frequency of 0.57 Hz, peaked at 19 hpf at 0.96 Hz, and gradually decreased to <0.1 Hz by 27 hpf. Starting at 21 hpf, touching either the head or the tail of the embryos resulted in vigorous coils. The coils accelerated with development, reaching a maximum speed of contraction before 48 hpf, which is near the time of hatching. After 27 hpf, touching the embryos, particularly on the tail, could induce partial coils (instead of full coils). At this time, embryos started to swim in response to a touch, preferentially to the tail. The swim cycle frequency gradually increased with age from 7 Hz at 27 hpf to 28 Hz at 36 hpf. Lesions of the central nervous system rostral to the hindbrain had no effect on the three behaviors. Lesioning the hindbrain eliminated swimming and touch responses, but not the spontaneous contractions. Our observations suggest that the spontaneous contractions result from activation of a primitive spinal circuit, while touch and swimming require additional hindbrain inputs to elicit mature locomotor behaviors. PMID- 9858264 TI - Fibroblast growth factor receptor signaling in Xenopus retinal axon extension. AB - Fibroblast growth factor receptors (FGFRs) and N-cadherin both regulate axon extension in developing Xenopus retinal ganglion cells (RGCs). Cultured cerebellar neurons have been shown to require FGFR activity for N-cadherin stimulated neurite outgrowth, raising the possibility that N-cadherin is a FGFR ligand. To investigate this possibility in the developing visual system, retinal neurons were transfected with a dominant-negative FGFR (XFD) and plated on purified N-cadherin substrates. XFD-expressing neurons extended markedly shorter processes than control GFP-expressing neurons, implicating a role for FGFRs in N cadherin-stimulated neurite outgrowth. To examine whether N-cadherin and FGFRs share the same pathway or use distinct second messenger pathways, specific inhibitors of implicated signaling molecules were added to neurons stimulated by N-cadherin, basic fibroblast growth factor (bFGF), or brain-derived nerve factor (BDNF) (which stimulates RGC outgrowth by a FGFR-independent mechanism). Diacylglycerol (DAG) lipase and Ca2+/calmodulin kinase II inhibitors both significantly reduced outgrowth stimulated by N-cadherin or bFGF but not by BDNF. Furthermore, we show that inhibiting DAG lipase activity in RGC axons extending in vivo toward the optic tectum reversibly slows axon extension without collapsing their growth cones. Thus, a common second-messenger signaling pathway mediating both N-cadherin- and bFGF-stimulated neurite extension is consistent with a model in which N-cadherin directly modulates the FGFR or a model whereby both FGFR and N-cadherin regulate the same second-messenger system. PMID- 9858265 TI - Intrinsic and synaptic properties of neurons in the vocal-control nucleus IMAN from in vitro slice preparations of juvenile and adult zebra finches. AB - A common theme of diverse neural systems is that circuits that are important for initial acquisition of learning do not necessarily serve as a substrate for the long-term storage of that memory. The neural basis of vocal learning in songbirds provides an example of this phenomenon, since a circuit that is necessary for vocal production during initial stages of vocal development apparently plays no subsequent role in controlling learned vocalizations. This striking functional change suggests the possibility of marked physiological changes in synaptic transmission within this circuit. We therefore examined intrinsic and synaptic properties of neurons in the cortical nucleus IMAN (lateral magnocellular nucleus of the anterior neostriatum), which forms part of this developmentally regulated circuit, in an in vitro preparation of the zebra finch forebrain. Although both functional and morphological characteristics of these neurons change substantially during vocal development, we did not observe widespread, substantive changes in the electrophysiological characteristics of juvenile versus adult IMAN neurons examined in vitro. Overall, both the intrinsic properties and synaptic responses of IMAN neurons were similar in slices from juvenile birds (at ages when lesions of IMAN disrupt vocal production) and in slices from adult birds (when IMAN lesions have no effect on song production). However, one intrinsic property that did vary between juvenile and adult cells was spike duration, which was longer in juvenile cells, suggesting the potential for activation of second-messenger cascades and/or enhanced synaptic transmission onto target cells of IMAN neurons. The pattern of synaptic response observed in both juvenile and adult cells suggests that IMAN projection neurons receive direct excitatory afferent inputs, as well as disynaptic inhibitory inputs from interneurons within IMAN. Activation of inhibitory interneurons rapidly curtails the excitatory response seen in projection neurons. This inhibition was abolished by bicuculline, indicating that the inhibitory interneurons normally exert their postsynaptic response via GABA(A) receptors on projection neurons. The inhibitory response could also be blocked by CNQX (6-cyano-7-nitroquinoxaline-2,3-dione), suggesting that the activation of inhibitory interneurons within IMAN may be governed primarily by AMPA receptors. PMID- 9858266 TI - Role for cell adhesion and glycosyl (HNK-1 and oligomannoside) recognition in the sharpening of the regenerating retinotectal projection in goldfish. AB - Cell-adhesion molecules (CAMs) are thought to play crucial roles in development and plasticity in the nervous system. This study tested for a role for cell adhesion and in particular, the recognition of two glycosyl epitopes (HNK-1 and oligomannoside) in the activity-driven sharpening of the retinotopic map formed by the regenerating retinal fibers of goldfish. HNK-1 is a prominent glycosyl epitope on many CAMs and extracellular matrix (ECM) molecules, including NCAM, L1, ependymin, and integrins, which have all been implicated in synaptic plasticity. To test for a role of HNK-1 in the sharpening process, we used osmotic minipumps to infuse HNK-1 antibodies for 7-21 days into the tectal ventricle starting at 18 days after optic nerve crush. Retinotopic maps recorded at 76-86 days postcrush showed a lack of sharpening similar to that seen previously with two antibodies to ependymin, an HNK-1-positive ECM component present in cerebrospinal fluid. The multiunit receptive fields at each point averaged 26 degrees versus 11-12 degrees in regenerates infused with control antibodies or Ringer's alone. The HNK-1 epitope also binds to the G2 domain of laminin to mediate neuron-ECM adhesion. To test for a role for laminin, a polyclonal antibody was similarly infused and also prevented sharpening to approximately the same degree. The results support a role for the HNK-1 epitope and laminin in retinotectal sharpening. The oligomannoside epitope (recognized by monoclonal antibody L3) on the CAM L1 interacts with NCAM on the same cell to promote stronger L1 homophilic interactions between cells. Both an L1-like molecule and NCAM are prominently reexpressed in the regenerating retinotectal system of fish. Infusion of oligomannosidic glycopeptides resulted in decreased sharpening, with multiunit receptive fields that averaged 22.7 degrees. Infusions of mannose-poor glycopeptides less prominently disrupted sharpening, with average multiunit receptive fields of 18 degrees. Thus, oligomannosidic glycans in particular may play a role in retinotopic sharpening. Blocking glycan-mediated interactions between CAMs and ECM molecules could decrease the extent of exploratory growth of retinal axon collaterals, preventing them from finding their retinotopic sites, or could interfere with L1 or NCAM and laminin binding at the synaptic densities preventing stabilization of retinotopically appropriate synapses. Together, these results support a prominent role for cell adhesion and glycan epitopes in visual synaptic plasticity. PMID- 9858267 TI - A transient role for ciliary neurotrophic factor in chick photoreceptor development. AB - Previous studies suggest that ciliary neurotrophic factor (CNTF) may represent one of the extrinsic signals controlling the development of vertebrate retinal photoreceptors. In dissociated cultures from embryonic chick retina, exogenously applied CNTF has been shown to act on postmitotic rod precursor cells, resulting in an two- to fourfold increase in the number of cells acquiring an opsin positive phenotype. We now demonstrate that the responsiveness of photoreceptor precursors to CNTF is confined to a brief phase between their final mitosis and their terminal differentiation owing to the temporally restricted expression of the CNTF receptor (CNTFR alpha). As shown immunocytochemically, CNTFR alpha expression in the presumptive photoreceptor layer of the chick retina starts at embryonic day 8 (E8) and is rapidly down-regulated a few days later prior to the differentiation of opsin-positive photoreceptors, both in vivo and in dissociated cultures from E8. We further show that the CNTF-dependent in vitro differentiation of rods is followed by a phase of photoreceptor-specific apoptotic cell death. The loss of differentiated rods during this apoptotic phase can be prevented by micromolar concentrations of retinol. Our results provide evidence that photoreceptor development depends on the sequential action of different extrinsic signals. The time course of CNTFR alpha expression and the in vitro effects suggest that CNTF or a related molecule is required during early stages of rod differentiation, while differentiated rods depend on additional protective factors for survival. PMID- 9858268 TI - Organizational effects of estrogens on brain vasotocin and sexual behavior in quail. AB - Reproductive behavior is sexually differentiated in quail: The male-typical copulatory behavior is never observed in females even after treatment with high doses of testosterone (T). This sex difference in behavioral responsiveness to T is organized during the embryonic period by the exposure of female embryo to estrogens. We showed recently that the sexually dimorphic medial preoptic nucleus (POM), a structure that plays a key role in the activation of male copulatory behavior, is innervated by a dense steroid-sensitive network of vasotocin immunoreactive (VT-ir) fibers in male quail This innervation is almost completely absent in the female POM and is not induced by a chronic treatment with T, suggesting that this neurochemical difference could be organizational in nature. This idea was tested by injecting fertilized quail eggs of both sexes on day 9 of incubation with either estradiol benzoate (EB) (25 microg, a treatment that suppresses the capacity to show copulatory behavior in adulthood) or the aromatase inhibitor R76713 (10 microg, a treatment that makes adult females behaviorally responsive to T), or with the solvents as a control (C). At 3 weeks posthatch, all subjects were gonadectomized and later implanted with Silastic capsules filled with T. Two weeks later, all birds were perfused and brain sections were processed for VT immunocytochemistry. Despite the similarity of the adult endocrine conditions of the subjects (all were gonadectomized and treated with T Silastic implants providing the same plasma level of steroid to all subjects), major qualitative differences were observed in the density of VT-ir structures in the POM of the different groups. Dense immunoreactive structures (fibers and a few cells) were observed in the POM of C males but not females; EB males had completely lost this immunoreactivity (and lost the capacity to display copulatory behavior); and, conversely, R76713 females displayed a male-typical VT ir system in the nucleus (and also high levels of copulatory behavior). Similar changes in immunoreactivity were seen in the nucleus of the stria terminalis and in the lateral septum (VT-ir fibers only in this case) but not in the magnocellular vasotocinergic system. These neurochemical changes closely parallel the effects of the embryonic treatments on male copulatory behavior. The vasotocinergic system of the POM can therefore be considered an accurate marker of the sexual differentiation of brain circuits mediating this behavior. PMID- 9858269 TI - Target neuron specification of short-term synaptic facilitation and depression in the cricket CNS. AB - We investigated the role of retrograde signals in the regulation of short-term synaptic depression and facilitation by characterizing the form of plasticity expressed at novel synapses on four giant interneurons in the cricket cercal sensory system. We induced the formation of novel synapses by transplanting a mesothoracic leg and its associated sensory neurons to the cricket terminal abdominal segment. Axons of ectopic leg sensory neurons regenerated and innervated the host terminal abdominal ganglion forming monosynaptic connections with the medial giant interneuron (MGI), lateral giant interneuron (LGI), and interneurons 7-1a and 9-2a. The plasticity expressed by these synapses was characterized by stimulating a sensory neuron with pairs of stimuli at various frequencies or with trains of 10 stimuli delivered at 100 Hz and measuring the change in excitatory postsynaptic potential amplitude recorded in the postsynaptic neuron. Novel synapses of a leg tactile hair on 7-1a depressed, as did control synapses of cercal sensory neurons on this interneuron. Novel synapses of leg campaniform sensilla (CS) sensory neurons on MGI, like MGI's control synapses, always facilitated. The form of plasticity expressed by novel synapses is thus consistent with that observed at control synapses. Leg CS synapses with 9-2a also facilitated; however, the plasticity expressed by these sensory neurons is dependent on the identity of the postsynaptic cell since the synapses these same sensory neurons formed with LGI always depressed. We conclude that the form of plasticity expressed at these synaptic connections is determined retrogradely by the postsynaptic cell. PMID- 9858270 TI - Ovarian Brenner tumors: MR imaging characteristics. AB - This study describes the appearance of Brenner tumors on MR imaging and compares quantitative signal intensity measurements of Brenner tumors with that of other ovarian tumors. A search of pathologic and MR records disclosed patients who had MRIs showing Brenner tumors prior to surgical excision. Patients (21) with other surgically proven ovarian masses were randomly selected for comparison. MR imaging was performed at 1.5 T with phased array multicoils and fast spin echo T2 weighted images. Region-of-interest measurements of signal intensity (SI) were made to calculate signal intensity ratios (SIR = mass SI/muscle SI). Brenner tumors showed significantly lower SIR than other tumors on T2-weighted images (p = 0.004) and similar SIR on T1-weighted images. Brenner tumors show lower signal intensity on T2-weighted images than other non-fibrous ovarian tumors. This lower signal intensity may result from the extensive fibrous content of these tumors. PMID- 9858271 TI - High-resolution cardiac imaging using an interleaved 3D double slab technique. AB - A three-dimensional (3D) gradient-echo sequence with interleaved double-slab excitation was developed and optimized for the requirements in pediatric cardiac imaging. For this purpose high contrast between blood and myocardium signal should be obtained without the use of contrast agents. An acceptable measuring time for a large region examined with high spatial resolution should be achieved as well, especially with regard to the small structures of the heart and vessels of infants. The presented approach works with gradient moment nulling and a short echo time of 5.5 ms resulting in generally high signal intensity and only minor signal losses due to turbulent flow. The sequence allows simultaneous ECG-gated recording of two separately excited slabs with small thickness (10 mm) and with a distance of several centimeters between them. Thus, common effects of presaturation in 3D imaging can be avoided, although a relatively short measuring time is achievable. In order to get a 3D data set with good signal homogeneity of blood and of the other structures across a large volume of interest several double-slab measurements with suitable positions must be performed. The latter aspect is especially important for postprocessing techniques as multiple planar reconstruction and maximum intensity projection. Examples of applications of the new technique and appropriately postprocessed images are presented allowing demonstration even of subtle cardiac malformations. PMID- 9858273 TI - Functional MRI of the motor cortex using a conventional gradient system: comparison of FLASH and EPI techniques. AB - Gradient echo (GE) and echo planar imaging (EPI) techniques are two different approaches to functional MRI (fMRI). In contrast to GE sequences, the ultra short EPI technique facilitates fMRI experiments with high spatial and temporal resolution or mapping of the whole brain. Although it has become the method of choice for fMRI, EPI is generally restricted to modern scanners with a strong gradient system. The aim of our study was to evaluate the applicability of EPI for fMRI of the motor cortex using a 1.5 T scanner with a conventional gradient system of 10 mT/m (rise time: 1 ms). Therefore, EPI was compared with a well established high-resolution fast low-angle shot (FLASH) technique (matrix size 1282). The FLASH technique was applied additionally with a 64(2) matrix size to exclude influences caused by different spatial resolution, because the EPI sequence was restricted to a 64(2) matrix size. A total of 35 healthy volunteers were included in this study. The task consisted of clenching and spreading of the right hand. FLASH and EPI techniques were compared regarding geometric distortions as well as qualitative and quantitative fMRI criteria: Mean signal increase between activation and rest and the area of activation were measured within the contralateral, ipsilateral, and supplementary motor cortex. The quality of subtraction images between activation and rest, as well as the quality of z-maps and time course within activated regions of interest, was evaluated visually. EPI revealed significant distortions of the anterior and posterior brain margins; lateral distortions (relevant for the motor cortex) could be neglected in most cases. The mean signal increase was significantly higher using FLASH 1282 compared to FLASH 64(2) and EPI 64(2), whereas the activated areas proved to be smaller in FLASH 1282 functional images. Both results can be explained by well-documented partial volume effects, caused by different voxel size. Similar quality of the subtraction images and of the time courses in different regions of interest were found for all techniques under investigation, but slightly reduced quality of z-map in FLASH 1282. Within the limits of reproducibility and measurement accuracy, the location of contralateral activation was similar using FLASH and EPI sequences. In conclusion, EPI proved to be a reliable technique for fMRI of the motor cortex, even on an MR scanner with a conventional gradient system. PMID- 9858272 TI - Reduced MTR in the corticospinal tract and normal T2 in amyotrophic lateral sclerosis. AB - The objective of this study was to test the hypothesis that magnetization transfer ratios (MTR) are decreased in the corticospinal tract of patients with amyotrophic lateral sclerosis (ALS); to determine if T2 is increased in corticospinal tract or reduced in motor cortex in ALS; to determine if corticospinal tract MTR correlates with a clinical measure of motor neuron function in ALS. Ten ALS patients and 17 age-matched controls were studied. Double spin echo MRI and 3D gradient echo MRI with and without off-resonance saturation were acquired on each subject. 3D data sets were coregistered and resliced to match the spin echo data set. MTR was calculated for corticospinal and non-corticospinal tract white matter. T2 was calculated for corticospinal and non-corticospinal tract white matter, motor cortex and non-motor cortex. MTR was reduced by 2.6% (p < .02) in corticospinal, but not in non-corticospinal, tract white matter in ALS. There was no difference in T2 in any brain region. The correlation between a clinical measure of motor neuron function and corticospinal tract MTR was statistically significant. These findings are consistent with the known pathology in ALS and suggest that MTR is more sensitive than T2 for detecting involvement of the corticospinal tract. Quantitative MTR of the corticospinal tract may be a useful, objective marker of upper motor neuron pathology in ALS. PMID- 9858274 TI - How does brain MRI lesion volume change on serial scans in patients with multiple sclerosis? AB - Although lesion load changes on conventional T2-weighted brain magnetic resonance imaging (MRI) scans from patients with multiple sclerosis (MS) are used to monitor the effect of treatment, there is no clear definition of how lesion load changes over years according to the lesion load present at a baseline evaluation. In the present study, we evaluated the relationship between lesion load changes over time and lesion load at a baseline evaluation in a group of untreated patients with MS. We scanned nineteen patients on two separate occasions with a mean interval 16.4 months between the two examinations. In each scanning session, a scan with forty contiguous 3-mm-thick axial slices was acquired. We assessed MRI lesion loads using a semi-automated local thresholding technique. Both a linear (p < 0.0001) and a quadratic component (p = 0.0008) of the baseline volume were significant in describing the follow-up volume. The equation to model this finding was as follows: Vf = beta0 Vb + beta1 (Vb)2, where Vf is the lesion volume at follow-up, Vb is the lesion volume at baseline, beta0 = 0.834 (SE = 0.098), and beta1 = 0.014 (SE = 0.003) (mL)(-1). Our data indicate that lesion volume changes detectable on serial brain MRI studies from patients with MS are dependent on the extent of lesion burden present on the baseline MRI scans. This finding has to be considered when planning phase III trials. PMID- 9858275 TI - Brain MRI lesion volume measurement reproducibility is not dependent on the disease burden in patients with multiple sclerosis. AB - We evaluated the potential effect of the lesion burden on the reproducibility of repeated lesion volume (LV) measurements from brain magnetic resonance imaging (MRI) scans of patients with multiple sclerosis (MS). Dual-echo, conventional spin echo brain MRI scans were obtained from 107 patients with MS. On proton density-weighted images, LV was assessed three times by the same raters, using a semi-automated, local thresholding technique for lesion segmentation. Mean LV (MLV) was 16.1 mL (range = 0.7-57.3 mL). The mean intra-observer coefficient of variation (COV) for the three measurement replicates was 2.6% (range = 0.2-7.2%). The intra-observer measurement variance (Var) increased with MLV and the fitted model was Var = 0.00187 MLV1.84. This indicates that LV measurements can be considered as measures whose variances are proportional to the square of their mean values, i.e., these measures have constant COV. Using a semi-automated, local thresholding segmentation technique, the reproducibility of LV measurements from brain MRI scans of patients with MS is not significantly influenced by varying lesion burdens. PMID- 9858276 TI - On- and off-resonance spin-lock MR imaging of normal human brain at 0.1 T: possibilities to modify image contrast. AB - The aim of the present investigation was to determine spin lock (SL) relaxation parameters for the normal brain tissues and thus, to provide basis for optimizing the imaging contrast at 0.1 T. 68 healthy volunteers were included. On-resonance spin lock relaxation time (T1rho) and off-resonance spin lock relaxation parameters (T1rho(off), Me/Mo), MT parameters (T1sat, Ms/Mo), and T1, T2 were determined for the cortical gray matter, and for the frontal and parietal white matters. The T1rho for the frontal and parietal white matters ranged from 110 to 133 ms and from 122 to 155 ms with locking field strengths from 50 microT to 250 microT, respectively. Accordingly, the values for the gray matter ranged from 127 to 155 ms. With a locking field strength of 50 microT, T1rho(off) for the frontal and parietal white matters were from 114 to 217 ms and from 126 to 219 ms, and for the gray matter from 136 to 267 ms with the angle between the effective magnetic field (B(eff)) and the z-axis (theta) ranging from 60 degrees to 15 degrees, respectively. The T1rho of the white and gray matters increased significantly with increasing locking field amplitude (p < 0.001). The T1rho(off) decreased significantly with increasing theta (p < 0.001). T1rho and T1rho(off) with theta > or = 30 degrees were statistically significantly shorter in the frontal than in the parietal white matters (p < 0.05). The duration, amplitude and theta of the locking pulse provide additional parameters to optimize contrast in brain SL imaging. PMID- 9858277 TI - Multicomponent water proton transverse relaxation and T2-discriminated water diffusion in myelinated and nonmyelinated nerve. AB - The influence of compartmental boundaries on water proton transverse relaxation and diffusion measurements was investigated in three distinct excised nerves, namely, the non-myelinated olfactory nerve, the Schwann cell myelinated trigeminal nerve, and the oligodendrocyte myelinated optic nerve of the garfish. The transverse relaxation decay curves were multiexponential and their decomposition yielded three primary components with T2 values approximately 30 50, 150, and 500 ms, which were subsequently assigned to water protons in the myelin, axoplasm, and interaxonal compartments. The short T2 component was absent in the non-myelinated olfactory nerve, but present in both myelinated nerves and thus provides supporting evidence for the use of quantitative T2 measurements to measure the degree of myelination. The signal contribution of each T2 component to the apparent diffusion coefficient measurements was varied by incrementing the spin-echo time with a preparatory CPMG train of radiofrequency pulses. The apparent diffusion coefficient and its anisotropy were shown to be independent of the spin-echo time over the range of 70 to 450 ms. PMID- 9858278 TI - Regional distribution of manganese found in the brain after injection of a single dose of manganese-based contrast agents. AB - Manganese (Mn) complexes are unstable and dissociate in vivo. Because of the release of this metal, there exists some concern about the potential long-term neurotoxicity associated with the use of Mn-based contrast agents. This latter problem arises because manganese is known to accumulate in specific regions of the brain of people intoxicated by this metal. It was previously demonstrated that Mn can accumulate in the mice brain after administration of 5 micromol/kg of MnCl2, Mn-diethylenetriaminepentaacetate (Mn-DTPA), or Mn-dipyridoxal diphosphate (Mn-DPDP). In order to better characterize the behavior of Mn complexes after administration, this study assesses the regional distribution of Mn in the brain after i.v. injection of a single dose of MnCl2 or Mn-DTPA. Male Wistar rats received an i.v. injection of 5 micromol/kg of 54Mn as MnCl2 or Mn-DTPA. The rats were killed at one and two weeks post exposure. The distribution of the radioactivity in the slices was monitored by autoradiography. For both MnCl2 or Mn-DTPA, we observed that the radioactivity was dispersed in the entire brain, but the radioactivity was higher in several regions. No difference was observed between MnCl2 or Mn-DTPA in the regional distribution of Mn, and no difference was observed between the two times of exposure (1 week or 2 weeks). The uptake of Mn was minimal in corpus callosum. Maximal Mn concentration was observed in the hippocampal region, thalamus, colliculi, amygdala, olfactory nuclei, and cerebellum. PMID- 9858279 TI - Statistical methods for detecting activated regions in functional MRI of the brain. AB - Two statistical tests for detecting activated pixels in functional MRI (fMRI) data are presented. The first test (t-test) is the optimal solution to the problem of detecting a known activation signal in Gaussian white noise. The results of this test are shown to be equivalent to the cross-correlation method that is widely used for activation detection in fMRI. The second test (F test) is the optimal solution when the measured data are modeled to consist of an unknown activation signal that lies in a known lower dimensional subspace of the measurement space with added Gaussian white noise. A model for the signal subspace based on a truncated trigonometric Fourier series is proposed for periodic activation-baseline imaging paradigms. The advantage of the second method is that it does not assume any information about the shape or delay of the activation signal, except that it is periodic with the same period as the activation-baseline pattern. The two models are applied to experimental echo planar fMRI data sets and the results are compared. PMID- 9858280 TI - Data-driven curvilinear reconstructions of 3D MR images: application to cryptogenic extratemporal epilepsy. AB - This paper presents a data-driven method for the reconstruction and visualisation of curvilinear slices from three-dimensional (3D) magnetic resonance (MR) scans of the head. Visualisation of curvilinear slices, rather than standard planar slices, produces symmetrical views of the cortex and allows small abnormalities to be detected by comparing the two hemispheres of the brain. In our method, the surface defined by the upper half of the brain is used as a reference shape for curvilinear reconstructions. The brain is first segmented from the 3D scan using a 3D region growing method associated to an unsupervised threshold selection technique. The upper half of the segmented brain is then extracted and fitted by a deformable surface model. This surface is finally interactively moved by the operator in the 3D scan, to visualise the desired curvilinear slice, which is projected on the screen as a two-dimensional image. We show an application of this visualisation technique to the localisation of cerebral epileptogenic lesions. The procedure has proven efficient and handy in clinical use. PMID- 9858281 TI - Two methods for semi-automated quantification of changes in ventricular volume and their use in schizophrenia. AB - Two semi-automated methods for quantification of ventricular volume change from baseline and follow-up magnetic resonance imaging scans have been developed. Technique 1 employs direct segmentation of the ventricles from both the scans using thresholding and contour extraction. Technique 2 operates on difference images produced by voxel based intensity subtraction of the baseline from the registered follow-up images. Here, all voxels with intensities above a noise threshold and in a restricted area are monitored to compute volumetric changes. In phantom measurements the first technique was accurate to 0.0046%, the second to 0.167% of the phantom volume. Results from normal volunteers was that the average ventricular volume changed by 1.52% and 1.54% for images acquired within 9 months using techniques 1 and 2, respectively. With schizophrenic patients mean change of 10.78% and 9.43% were found employing the first and second procedures, respectively. All measurements agreed with a radiologist's visual grading of the changes. Robust, objective, fast, easy-to-use, and fairly accurate procedures have been developed and validated to quantify volumetric changes. PMID- 9858282 TI - In vivo echo-planar imaging of rat spinal cord. AB - An integrated approach to echo-planar imaging of rat spinal cord in vivo with a small field of view (FOV) is presented. This protocol is based on a multishot interleaved echo-planar imaging (EPI) sequence and includes: 1) use of an inductively coupled implantable coil for improved signal-to-noise ratio (SNR); 2) three-dimensional (3D) automatic shimming of the magnetic field over the spinal cord; and 3) post-acquisition data processing using a multireference scan for minimizing image artifacts. Some of the practical issues in implementing this protocol are discussed. This imaging protocol will be useful in characterizing the spinal cord pathology using techniques that are otherwise time-consuming, such as diffusion tensor imaging. PMID- 9858283 TI - Experimental evaluation of nonlinearities of small-sized insertable gradient coils. AB - A phase imaging technique is proposed to map out and quantify gradient nonlinearities of small-sized insertable gradient coils, assuming the whole-body system gradients are highly linear in the domain of interest. The theory is developed and simple equations are derived to allow quantification. It is applied to a 4-loop 18-cm diameter cylindrical gradient coil of optimal design. Experimental gradient nonlinearity maps are obtained for different fields of view. Gradient non-linearities are quantified locally and in regions of interest, showing close agreement with model data. PMID- 9858284 TI - Magnetic resonance spectroscopic imaging for visualization and correction of distortions in MRI: high precision applications in neurosurgery. AB - We present a method for the quantification and correction of geometrical/intensity distortions of magnetic resonance images predominantly caused by bulk magnetic susceptibility shifts due to susceptibility heterogeneities of measured biologic tissues and shape of the object under investigation. The method includes precise and fast measurements of the static magnetic-field distribution inside the measured object and automated data processing. Magnetic-field deviations in the range (-2.4; 2.6) ppm were found in the human brain at B0 = 1.5 T. For routinely used imaging parameters, with a read gradient strength of about approximately 1 mT/m, the magnetic-field perturbations in the human brain can cause geometrical distortions up to +/-4 mm and intensity changes up to +/-50%. MR images corrected by the described method are suitable for planning high precision applications in neurosurgery. PMID- 9858285 TI - 1H MR spectroscopy monitoring of changes in choline peak area and line shape after Gd-contrast administration. AB - Fifteen percent loss in the peak area of choline containing compounds (Cho) was recently observed in 1H MR spectra of contrast-enhancing tumor at 5-10 min after Gd-contrast administration [Magn. Reson. Med. 37:222-225, 1997]. In this study, chemical shift imaging (CSI, 1500/135 ms PRESS) was used to assess the spectral changes in 47 Gd-enhancing glial brain tumors and metastatic brain tumors measured at 0-5, 5-10, and/or 10-15 min after administration of Gd-contrast. Percent Cho peak area losses measured at these times, 3 +/- 3, 12 +/- 2, and 14 +/- 3 SEM, respectively, coincided with trends of line narrowing and up-field shift of the Cho peak. Significant changes in creatine and N-acetyl acetate signals were not observed. It is concluded that the Gd-induced loss of tumor Cho signal measured after 5 min, typically required for post contrast-MRI and the positioning of the CSI volume on tumor, shows little further change with time, if any. PMID- 9858286 TI - In vivo differential diagnosis of prostate cancer and benign prostatic hyperplasia: localized proton magnetic resonance spectroscopy using external-body surface coil. AB - Localized proton-stimulated echo acquisition mode (STEAM) spectroscopy was performed in seven patients with benign prostatic hyperplasia (BPH), six patients with prostate cancer, and seven healthy volunteers to determine whether citrate levels detected using a saddle-type external-body surface coil (two loops of 13 cm x 17 cm) could reliably discriminate BPH from prostatic cancer. Relative area ratios of citrate level to choline plus creatine or citrate to lipid signal were compared with postoperative pathologic histology findings. The metabolic signals were well detectable as much as the line width of water resonance was ranging from 5 to 9 hz. Average SNRs of citrate in BPH and prostate cancer were 11.4 and 1.9, respectively. The major finding was consistently lower citrate levels in prostate cancer compared with BPH and normal prostate central gland. This was significantly (p < 0.01) reflected by lower mean citrate/[creatine+choline] peak area ratio and citrate/lipid peak area ratio observed for region of cancer (0.446 +/- 0.063, 0.097 +/- 0.030) compared with BPH (1.458 +/- 0.107, 0.786 +/- 0.162) and normal central gland (1.418 +/- 0.129, 0.175 +/- 0.011), respectively. These studies demonstrate the potential of citrate spectrum detected by an external body surface coil as an in vivo marker for discriminating prostate cancer from BPH. PMID- 9858287 TI - 13C imaging by double resonance scalar-coupling editing. AB - The two-dimensional FT Imaging of 13C-glucose obtained with twin spin-echo double resonance sequence is presented. The images have been obtained by a doubly tuned surface coil on samples containing water and 13C-enriched glucose in water. It is shown, both theoretically and experimentally, that the whole editing capability of the twin spin-echo double resonance imaging sequence is also preserved in the presence of the radiofrequency field inhomogeneity produced by the surface coil. As in an efficient selective irradiation method, the enhancement in the signal-to noise ratio with respect to the direct 13C detection, depends on the number of protons J-coupled to 13C. PMID- 9858288 TI - Diagnosis of spine metastases by FDG imaging using a gamma camera in the coincidence mode. AB - PURPOSE: To illustrate the detection of metastatic lung cancer using a conventional dual detector gamma camera modified to operate in the coincidence mode to image the static distribution of 2-deoxy-2-[F-18]fluoro-D-glucose (FDG). METHODS: A patient with known lung cancer underwent FDG imaging to evaluate the extent of metastatic disease. Twenty-three-minute emission image acquisition at one bed position using a dual detector coincidence gamma camera was performed to evaluate the entire chest and upper abdomen. RESULTS: In addition to detecting the malignant lung nodule, mediastinal lymph node metastases and a rib metastasis, symptomatic metastases to lower thoracic and upper lumbar vertebral bodies, which were not identified on radiographs, CT, or bone scan, were shown clearly by FDG coincidence gamma camera imaging. CONCLUSION: FDG tumor imaging in the body is feasible using a dual detector gamma camera operating in coincidence mode and may find an important role in a certain subset of FDG tumor imaging. PMID- 9858289 TI - A prospective comparison of clinical examination, MRI, bone SPECT, and arthroscopy to detect meniscal tears. AB - Recent studies have shown that SPECT bone scintigraphy is valuable to detect meniscal tears of the knee. This has not been formally assessed in a prospective study, and no substantive study has compared bone SPECT with other noninvasive diagnostic methods. One hundred consecutive patients referred to an orthopedic surgeon with undiagnosed knee pain were assessed by clinical examination, MRI, SPECT bone scintigraphy, and arthroscopy. The MRI and SPECT bone scan findings were reported blinded to other information. Using arthroscopy as a gold standard, both MRI and SPECT showed high diagnostic ability to detect meniscal tears, with respective sensitivity rate, specificity rates, and positive and negative predictive accuracies of 80%, 71%, 84%, and 71% for MRI and 84%, 80%, 88%, and 76% for SPECT. Some meniscal tears were detected by MRI alone (n = 5) or SPECT alone (n = 8). SPECT bone scintigraphy is a suitable alternative to MRI to detect meniscal tears. The comparable diagnostic ability of SPECT bone scintigraphy implies that it can be used successfully when MRI is unavailable or unsuitable. PMID- 9858290 TI - Dual isotope F-18 FDG and Tc-99m RBC imaging for lung cancer. AB - PURPOSE: To produce fluorodeoxyglucose (FDG) images with markers for normal organs and large blood vessels in patients with suspected lung cancer. METHODS: Dual isotope SPECT images were made using F-18 FDG- and Tc-99m-labeled autologous red cells. RESULTS: FDG-positive lesions are localized in relation to major structures when they are viewed in a fused rotating three-dimensional display or in cross sections. CONCLUSIONS: Tc-99m red cell and FDG fusion imaging provides relative location information for lung tumors. PMID- 9858291 TI - Time interval between the last dose of propylthiouracil and I-131 therapy influences cure rates in hyperthyroidism caused by Graves' disease. AB - PURPOSE: To examine the effects of propylthiouracil (PTU) pretreatment on the outcome of initial I-131 therapy for Graves' disease. DESIGN: A retrospective chart review was done. PATIENTS AND METHODS: The authors studied 106 patients in an outpatient nuclear medicine setting who were given initial I-131 therapy for Graves' disease from September 1989 to March 1993 and followed for at least 6 months after therapy. These patients were divided into groups based on whether they had ever received PTU or, if they had received PTU, the length of time between the last dose of PTU and the I-131 therapy dose. Measured failure rates of initial I-131 therapy were based on recurrent or continued hyperthyroidism. RESULTS: Treatment failure rates increased markedly from 2.5% in non-PTU-treated patients (n = 80) to 23.1% (n = 26) in patients pretreated with PTU (P = 0.003). Although not significant, two PTU-pretreated subgroups showed a trend toward increased failure rates. The failure rate was 15.4% (n = 13) in patients whose last dose of PTU was 7-14 days before I-131 therapy, and it increased further to 30.8% (n = 13) in patients whose last dose of PTU was within 1 week of I-131 therapy. CONCLUSIONS: PTU pretreatment within 2 weeks of I-131 treatment is a strong independent risk factor in failure rates after initial I-131 therapy in patients with Graves' disease. Patients should be free of PTU for 2 weeks before I-131 therapy if they are able to tolerate it, otherwise the dose of I-131 may need to be adjusted upward to diminish the risk that the initial I-131 therapy will fail. PMID- 9858293 TI - Altered radiopharmaceutical distribution in myeloma: bone tracer in the heart and heart tracer in the bone. AB - Tissue deposits of amyloid may lead to various clinical symptoms and signs, depending on the site of deposition. In addition, a specific subtype of amyloidosis, AL protein type, is associated with multiple myeloma. Two cases representing contrasting examples of altered radiopharmaceutical biodistribution in myeloma and amyloidosis are presented and discussed, with a bone tracer depicting the heart and a heart tracer depicting the bones. PMID- 9858292 TI - Can the C-14 urea breath test replace follow-up endoscopic biopsies in patients treated for Helicobacter pylori infection? AB - BACKGROUND: The C-14 urea breath test (UBT) is the most specific noninvasive test to detect Helicobacter pylori, with reported sensitivity and specificity rates of 90% and 95%, respectively. This test has not been evaluated for eradication after a therapeutic trial. The goal of this study was to assess the accuracy of C-14 UBT in the diagnosis and eradication of H. pylori infection in patients with duodenal ulcer who were treated with a triple drug regimen. METHODS: Sixty patients with active duodenal ulcers who tested positive for the rapid urease test had a C-14 UBT at 0 weeks (at enrollment) and at 6 and 12 weeks using 5 microCi (185 KBq) of C-14 urea. A single breath sample was collected at 15 minutes for UBT. H. pylori was eradicated using lansoprazole and two antibiotics. RESULTS: Receiver operator characteristic curves showed that, using a value of 400 counts per minute (cpm), UBT had a sensitivity rate of 91%, specificity rate of 93%, positive predictive value of 77%, and a negative predictive value of 97% in the prediction of H. pylori eradication. The mean + 3 SD of H. pylori-negative patients was 380.1 cpm; at this cutoff value, the sensitivity and specificity rates were 91.3% and 92.8%, respectively. CONCLUSION: The C-14 UBT was an accurate, rapid, and easily administered test to diagnose initial H. pylori infection and to monitor its eradication, thereby obviating the need for repeated endoscopic biopsies. PMID- 9858294 TI - Metastatic pulmonary, gastric, and renal calcification demonstrated on bone scintigraphy in a patient with malignant melanoma and renal failure. AB - PURPOSE: This case report describes the clinical, scintigraphic, and pathologic findings in a patient with an unexpected finding of a cutaneous malignant melanoma. METHOD: Multiple imaging studies were done, as was a pathologic examination of a suspicious pigmented lesion on the patient's back. RESULT: A Tc 99m MDP bone scan showed diffuse uptake in the skeleton, lungs, kidneys, and stomach. CONCLUSION: Metastatic calcification, as shown by isotope scintigraphy, is an unusual manifestation of metastatic cancer from a primary cutaneous melanoma. PMID- 9858295 TI - Identification of new renal scarring in repeated episodes of acute pyelonephritis using Tc-99m DMSA renal SPECT. AB - PURPOSE: This prospective study compared the value of Tc-99m DMSA renal planar scintigraphy with SPECT to detect new renal involvement in patients with repeated episodes of acute pyelonephritis (APN). MATERIALS AND METHODS: Children with suspected APN were transferred to our department for DMSA renal scans. Seventy two children (ages 1 week to 15 years) had DMSA planar and SPECT imaging performed twice because of clinical or laboratory suspicion of repeated APN. In addition, radiographic voiding cystourethrography was also performed in all cases. The presence of vesicoureteral reflux (VUR) was graded on a scale of 0 to 5. RESULTS: New lesions were observed with SPECT in 56 kidneys and with planar scintigraphy in 38 kidneys. No patients had a negative result of Tc-99m DMSA renal SPECT who also had a positive Tc-99m DMSA planar result. The degree of VUR as related to APN was diagnosed better with SPECT than with planar scintigraphy (46 compared with 30 and 10 compared with 8, respectively). There is a significant difference (P < 0.05) between the diagnostic ability of these two methods to identify the increased tendency of repeated APN to occur with high grade VUR compared with low-grade or no VUR. CONCLUSIONS: High-grade VUR is more commonly associated with renal injury than is low-grade or no VUR. If only Tc-99m DMSA renal planar scintigraphy is performed, renal scarring may be underestimated. Our results suggest that Tc-99m DMSA renal SPECT, rather than planar scintigraphy, should be used routinely in children with a clinical suspicion of APN, especially for those with scarred kidneys and high-grade VUR. PMID- 9858296 TI - Tl-201 myocardial SPECT in patients with systemic arterial diseases. AB - We compared the results of Tl-201 myocardial SPECT of patients with arteriosclerosis obliterans (ASO), abdominal aortic aneurysm (AAA), and Takayasu arteritis (TA), so that coronary artery disease could be evaluated in them. Twenty-three patients (ASO, 9; AAA, 8; TA, 6) had Tl-201 myocardial SPECT with stress testing, and SPECT data was assessed qualitatively and quantitatively. Qualitative analysis showed that myocardial perfusion was decreased most in patients with ASO, followed by patients with AAA, and was minimally decreased in patients with TA. Quantitative analysis indicated that segmental uptake was significantly less in four segments in ASO compared with TA, less in two segments in AAA compared with TA, and less in two segments in ASO compared with AAA. Thus patients with ASO and AAA were found to have a marked abnormality in Tl-201 myocardial SPECT, indicating that this procedure should be performed in these patients. In patients with TA, however, because of minimal abnormality in myocardial perfusion, the role of SPECT is limited. PMID- 9858297 TI - Proliferative myositis: a case of a pseudomalignant process. AB - A case of proliferative myositis, an unusual inflammatory pseudotumor of a skeletal muscle, is presented. Often it is erroneously misdiagnosed as a malignant neoplasm. PMID- 9858298 TI - Intraoperative localization of jejunal bleeding due to Dieulafoy's disease using Tc-99m RBC. PMID- 9858299 TI - Detection of malignant thymoma during myocardial perfusion tomography with Tc-99m sestamibi: potential implications for tumor evaluation and staging. PMID- 9858300 TI - Achalasia seen on I-123 thyroid scintigraphy. PMID- 9858301 TI - Early duodenogastric reflux in Tc-99m tetrofosmin myocardial scintigraphy: a case report. PMID- 9858302 TI - Porcelain gallbladder detected on bone scan. PMID- 9858303 TI - In-111 octreotide lung uptake in hypersensitive pneumonitis caused by fungal infection. PMID- 9858304 TI - Intrathoracic stomach causing a pitfall on thyroid imaging. PMID- 9858305 TI - Subcutaneous fat necrosis on gallium scan after abdominal surgery. PMID- 9858306 TI - Tc-99m tetrofosmin uptake in pulmonary tuberculosis with Pott's disease. PMID- 9858307 TI - Biliary and liver abscesses demonstrated with Tc-99m DISIDA and Ga-67 imaging. PMID- 9858308 TI - Decreased bilateral frontal lobe perfusion in dementia of amyotrophic lateral sclerosis. PMID- 9858309 TI - Visualization of a photopenic lesion secondary to a giant small bowel diverticulum with fecal retention during Meckel's scintigraphy. PMID- 9858310 TI - Ga-67 uptake in the aorta in Wegener's granulomatosis: overlap with Takayasu's arteritis? PMID- 9858311 TI - Current readings in nuclear medicine. PMID- 9858312 TI - Analysis of the human and mouse promoter region of the non-Hodgkin's lymphoma associated CD30 gene. AB - To investigate the regulation of CD30 at the level of transcription, we have isolated and compared the promoter sequence of human and murine CD30. Analysis of the human and mouse promoter identified a number of potential transcription factor binding sites, including ETS, MZF, AP-1, IK2, CREB, Stat, USF, and Spl. The absence of TATA or CAAT boxes and the identification of one major and three minor transcription initiation sites for CD30 suggest that it is a member of the class of TATA-less promoters that use initiator elements to correctly position the RNA polymerase. Comparison of the murine and human CD30 promoters identified a number of highly conserved regions, including an Spl site 40 bp upstream from the major start site and a downstream promoter element (DPE) that may be involved in directing transcriptional initiation of the CD30 gene. Functional analysis of the human CD30 promoter in transfected Jurkat T cells provided further evidence that these conserved regions are important regulatory elements in the CD30 promoter. PMID- 9858314 TI - Enhanced interleukin-2 gene transfer immunotherapy of breast cancer by coexpression of B7-1 and B7-2. AB - The capability of B7-1 to augment the antitumor activity of some cytokines has been shown primarily for such cytokines as interleukin-12 (IL-12), IL-7, and to a lesser extent IL-2. In this study, we investigate the ability of B7-1 and B7-2 to augment the antitumor activity of IL-2. Considering the affinity of both molecules for CD28 (T cell receptor for B7-1 and B7-2), we postulated that their potential to augment IL-2 antitumor activity would be similar. Two murine transgenic adenocarcinoma models were chosen to investigate the activity of adenoviral vectors constructed to express either B7-1 and IL-2 or B7-2 and IL-2. Before administering the vector intratumorally to tumor-bearing mice, we determined the expression of B7-1, B7-2, MHC I, and MHC II on these tumor cells and demonstrated positive expression of only MHC I. Intratumoral injection of the vector expressing B7-1 and IL-2 resulted in complete regression of all tumors treated. In contrast, the vector expressing B7-2 and IL-2 was significantly less effective at regressing PyMT tumors, whereas both double recombinant vectors demonstrated similar levels of complete regression in the Neu (NDL 8142) model. Regressed mice were all protected for rechallenge in both models and demonstrated antigen-specific cytotoxic T lymphocytes (CTL) in the PyMT model. These findings indicate that the combination of IL-2 with B7-1 or B7-2 significantly enhances the antitumor activity of IL-2. PMID- 9858313 TI - 16-Epiestriol, a novel anti-inflammatory nonglycogenic steroid, does not inhibit IFN-gamma production by murine splenocytes. AB - All the steroidal anti-inflammatory drugs currently available are glucocorticoids. The desired anti-inflammatory activities of glucocorticoids frequently are accompanied by adverse side effects, notably glycogenic activities and profound immunosuppression, that can limit clinical use. We recently identified 16-epiestriol, a naturally occurring steroid, as exhibiting significant anti-inflammatory activity without glycogenic activity. In the present study, we compared the effects of 16-epiestriol and hydrocortisone on the capacity of murine splenocytes to produce interferon-y (IFN-y). We injected young adult male BDF1 mice once with 20 mg/kg hydrocortisone or 20, 5, or 1 mg/kg 16 epiestriol and 4 h later harvested the splenocytes. Flow cytometric analysis confirmed that 16-epiestriol did not alter the number of CD3+ T cells in the spleen. In contrast to the suppressive effects of hydrocortisone, none of the 16 epiestriol concentrations inhibited concanavalin A-stimulated IFN-gamma production by spleen cells, as determined by ELISA. Incubating spleen cells from untreated mice in concentrations of 16-epiestriol ranging from 1 mg/ml to 100 pg/ml did not alter profiles of IFN-gamma production, in contrast to the suppressive dose-response effects of hydrocortisone. Collectively, these results support the contention that 16-epiestriol may be a clinically useful safe anti inflammatory steroid without profound immunosuppressive activities. PMID- 9858315 TI - Differential effects of IFN-beta1b on the proliferation of human vascular smooth muscle and endothelial cells. AB - The effect of human interferon (IFN)-beta1b (Betaseron) on the proliferation of cultured human vascular smooth muscle and endothelial cells was tested in vitro. IFN-beta1b inhibited thymidine incorporation and growth of primary cultures of human aortic and coronary artery smooth muscle in a concentration-dependent manner. The same concentrations of IFN-beta1b did not inhibit thymidine incorporation or growth of primary cultures of human aortic or coronary artery endothelial cells. IFN-beta1b induced the expression of MxA (an antiviral protein induced by type I IFNs) in both smooth muscle and endothelial cells, suggesting that both cell types express receptors for type I IFNs. The growth-inhibitory effect of IFN-beta1b could be mimicked by commercially available human IFN-beta, but not by IFN-alpha2 or IFN-alpha8. The effect of IFN-beta1b was species specific, as it did not inhibit thymidine incorporation in aortic smooth muscle cells derived from pig, rabbit, rat, or mouse. The action of IFN-beta1b on smooth muscle cells persisted for at least 4 days following a 24 h preincubation with IFN-beta1b. Human vascular smooth muscle cells treated with IFN-beta1b did not release lactate dehydrogenase, nor did they show any morphologic change, suggesting that IFN-beta1b was not toxic to the human vascular smooth muscle cells. IFN-beta1b inhibited vascular smooth muscle growth while having no growth inhibitory effect on endothelial cells obtained from the same blood vessel, making it a potential candidate for treating pathologic conditions where abnormal vascular smooth muscle proliferation is implicated, such as restenosis following balloon angioplasty or smooth muscle proliferation following vascular stenting. PMID- 9858316 TI - Induction of interferon synthesis and activation of interferon-stimulated genes by liposomal transfection reagents. AB - Liposome-mediated transfection is a widely used technique for the introduction of exogenous DNA into mammalian cells. We observed a significant induction of endogenous interferon (IFN)-stimulated genes (ISGs) in cells treated with the liposomal reagents, lipofectamine and DOSPER, in the absence of DNA. Liposome treatment induced expression of reporter constructs driven by IFN-responsive promoter elements, demonstrating a generalized effect on ISG expression. The kinetics of ISG induction were markedly delayed in response to liposome as compared with IFN or double-stranded RNA. ISG induction could be transferred to naive cells with conditioned medium from liposome-treated cells, suggesting that a secreted factor was responsible for this activity. A cell line defective in IFN signaling was refractory to liposome-induced ISG expression, indicating a role for IFN in this induction. Indeed, liposome treatment directly induced IFN-beta gene expression and, thus, represents a novel IFN inducer. IFN induction by liposomal reagents and its potential effects on transgene expression should be considered in the choice of transfection reagent. The ability of liposomal gene delivery reagents to induce IFN synthesis in the host may prove useful in gene therapy approaches to viral and neoplastic diseases. PMID- 9858317 TI - Impact of RNase L overexpression on viral and cellular growth and death. AB - The biologic actions of interferons (IFNs) are complex and involve multiple biochemical mechanisms, including the 2-5A system, a regulated RNA decay pathway. The 2-5A system is implicated in the antipicornavirus activity of IFN and in the control of apoptosis. To further investigate involvement of the 2-5A system in the control of viral and cellular growth and death, human RNase L cDNA was stably expressed in murine 3T3 cells from a constitutive cytomegalovirus (CMV) promoter. A clonal cell line, 3T3/pLZ, was isolated that overexpressed RNase L by >100-fold compared with levels of the endogenous murine RNase L. Interestingly, human RNase L levels in 3T3/pLZ cells decreased 3-fold as cells entered a confluent, growth arrest state, suggesting autoregulation. Overexpression of human RNase L greatly enhanced both the cell growth inhibitory activity of IFN and the proapoptotic activity of staurosporine. Furthermore, high levels of RNase L suppressed the replication of diverse viruses: encephalomyocarditis virus, vesicular stomatitis virus, human parainfluenza virus-3, and vaccinia virus. Additional reductions in viral growth were obtained by treating 3T3/pLZ cells with IFN (a + beta) before infections. These results directly demonstrate the anticellular and antiviral potential of the 2-5A system. PMID- 9858318 TI - Serum intercellular adhesion molecule-1 and long-term response to IFN-alpha2b therapy in chronic hepatitis C. AB - We have attempted to correlate the outcome of interferon (IFN) therapy with circulating soluble intercellular adhesion molecule-1 (sICAM-1) and the level of viremia in a sample of patients with chronic hepatitis C virus (HCV) infection. Forty-two patients were studied. Eighteen patients were maintained in long-term remission following IFN therapy, whereas 24 did not respond or relapsed. Serum concentrations of sICAM-1 were measured by enzyme-linked immunoassay. Viremia was measured by branched DNA signal amplification assay. Basal sICAM-1 was significantly higher in long-term responders than in nonresponder/relapsing patients. It was found that very high levels (>1000 ng/ml) were closely associated with long-term clinical response. A quantitative evaluation of viremia in basal conditions, which was significantly lower in long-term responders, gave completely opposite results. During treatment, sICAM-1 concentrations significantly decreased in the group of long-term responders but not in the nonresponders. sICAM-1 reduction was apparent as early as 1 month after treatment started. Serum sICAM-1 may be a useful parameter in evaluating the outcome of patients with chronic hepatitis C infection treated with IFN. PMID- 9858319 TI - Comparative study of the pharmacodynamic and pharmacologic effects of Betaseron and AVONEX. AB - The aim of this 1 week study was to compare the biologic effects induced by Betaseron and AVONEX using their approved dose, route, and schedule. Sixteen healthy volunteers were randomly assigned to receive either a single i.m. dose of AVONEX (6 million International Units [MIU]) or, every other day s.c. doses of Betaseron (8 MIU). Common side effects associated with interferon-beta (IFN-beta) treatment and biologic response parameters (neopterin, beta2-microglobulin, interleukin-10 [IL-10], and MxA protein levels in blood) were measured. Ibuprofen was administered to all subjects throughout the study. Fever, chills, and myalgia occurred most frequently and with the greatest severity between 6 and 12 h after the first dose of either IFN-beta. Despite the additional dosing of subjects in the Betaseron group, the incidence, duration, and severity of the side effects were not significantly different from those in the AVONEX group. Biologic response parameters reached similar maximum concentrations in both treatment groups. In the Betaseron group, neopterin and beta2-microglobulin levels remained significantly greater than baseline throughout the 7 day study, whereas those in the AVONEX group were elevated only through day 5. Betaseron treatment significantly increased IL-10 levels above baseline, but AVONEX treatment did not. The overall induction of neopterin, beta2-microglobulin, and IL-10 (as measured by area under the concentration-time curve) was significantly greater in the Betaseron group than the AVONEX group (p = 0.031). The results of this study demonstrate that the approved Betaseron dosing regimen, in combination with ibuprofen use, provided a significantly greater and more consistently elevated biologic response compared with that of AVONEX and did so with a side effects profile comparable to that of once a week AVONEX dosing. PMID- 9858320 TI - Murine GBP-2: a new IFN-gamma-induced member of the GBP family of GTPases isolated from macrophages. AB - We have cloned a new member of the interferon (IFN)-induced guanylate-binding protein (GBP) family of GTPases, murine GBP-2 (mGBP-2), from bone marrow-derived macrophages. mGBP-2 is located on murine chromosome 3, where it is linked to mGBP 1. With the identification of mGBP-2 there are now two human and two murine GBPs. Like other GBPs, mGBP-2 RNA and protein are induced by IFN-gamma. In addition, mGBP-2 shares with the other GBPs important structural features that distinguish this family from other GTPases. First, mGBP-2 contains only two of the three consensus sequences for nucleotide binding found within the classic GTP binding regions of other GTPases. A second amino acid motif found in mGBP-2 is a potential C-terminal site for isoprenoid modification, called a CaaX sequence. mGBP-2 is prenylated, as detected by [3H]mevalonate incorporation, when expressed in COS cells and preferentially incorporates the C-20 isoprenoid geranylgeraniol. Surprisingly, despite having a functional CaaX sequence, mGBP-2 is primarily cytosolic. GBP proteins are very abundant in IFN-exposed cells, but little is known about their function. mGBP-2 is expressed by IFN-gamma-treated cells from C57Bl/6 mice, whereas mGBP-1 is not. Thus, the identification of mGBP-2 makes possible the study of GBP function in the absence of a second family member. PMID- 9858321 TI - IP-10 gene transcription by virus in astrocytes requires cooperation of ISRE with adjacent kappaB site but not IRF-1 or viral transcription. AB - Transcription of the IP-10 gene requires interferon (IFN)-stimulated response element (ISRE) and kappaB sites to be induced by lipopolysaccharide (LPS), IFN gamma, virus, and poly(I:C). A requirement for Stat1 binding to ISRE for IFN gamma and IFN regulatory factor-1 (IRF-1) binding to ISRE for LPS, poly(I:C), and virus has been reported. We investigated whether viral transcription is required for IP-10 induction and how ISRE interacts with IRF-1 and with two kappaB sites. IP-10 mRNA was induced by Newcastle disease virus and Sendai virus in rat astrocytes and the human astrocytoma U251 cell line. IP-10 was also induced by UV irradiated virus, which is unable to carry out viral transcription. The minimal IP-10 virus response element (VRE) consists of an ISRE and adjacent kappaB site between -236 and -153, to which p50/p65 NF-kappaB proteins and IRF-like proteins bind. Virus induced NF-kappaB binding to an isolated kappaB sequence adjacent to ISRE. However, no protein binding to isolated ISRE was induced by virus. Virus also induced IP-10 in cells expressing a defective IRF-1 gene. Therefore, effective ISRE activity of IP-10 VRE may require an IRF-like protein binding, which is enhanced by an NF-kappaB heterodimer binding to an adjacent KB site. IRF 1 is not required for virus-induced IP-10 gene expression. PMID- 9858322 TI - Transcriptional activation and redox regulation of the tumor necrosis factor alpha promoter in human T cells: role of the CRE/kappa3 promoter region. AB - Aberrancies in T cell expression of tumor necrosis factor-alpha (TNF-alpha) are frequently observed in inflammatory states characterized by oxidative stress due to excessive generation of hydrogen peroxide (H2O2) and other reactive oxygen species (ROS). In this study, we examined the possible effects of oxidative stress on the expression of TNF-alpha protein and transcriptional activation of the TNF-alpha promoter in human T cells. Results show that exposure of resting T cells to micromolar concentrations of H2O2 did not induce TNF-alpha protein production or transcriptional activation of the TNF-alpha promoter. However, oxidative signals resulted in a dose-dependent suppression of TNF-alpha protein production and transcriptional activation in T cells stimulated with the lectin phytohemagglutinin (PHA) and phorbol myristate acetate (PMA). Optimal suppression of TNF-alpha promoter activity was observed when cells were exposed to oxidative stress during early T cell activation, and other experiments demonstrated that the transactivation responses of the TNF-alpha promoter were quite susceptible to inhibition by both oxidative and reducing changes in cellular redox. Furthermore, reporter gene assays with 5' deletion mutants of the TNF-alpha promoter showed that the CRE/kappa3 composite site played a major role in activation of the TNF alpha promoter by dual stimulatory signals and suppression of the TNF-alpha promoter by oxidative signals. Thus, T cell expression of TNF-alpha at the protein and transcriptional levels is highly regulated by changes in cellular redox, and the CRE/kappa3 composite site is important for both activation and redox regulation of the TNF-alpha promoter. PMID- 9858323 TI - Prostate cancer and black men. AB - Prostate cancer is a significant cause of death among men of all races in the United States, and it does disproportionately affect Black men. This disease poses a number of questions that desperately need answers. These questions involve not just the cause and prevention of the prostate cancer, but a very real and valid question is "does screening for and aggressive treatment of prostate cancer save lives." Almost all questions in prostate cancer are not questions unique to blacks or whites, or any specific population. These questions can only be answered through well-designed basic and clinical research studies. This research must be supported by both physician and patient participation. Conveying truthful, accurate information in this disease in which so much is unanswered is imperative. In American medical history, black men have often been misled or misinformed oftentimes by well-meaning paternalistic individuals. Physicians and laymen teaching about this disease must themselves realize and then truthfully convey "what is known, what is not known, and what is believed." This will allow the layman to make educated decisions regarding screening, treatment, and participation in clinical studies. PMID- 9858324 TI - The epidemiology of prostate cancer part I: descriptive epidemiology. AB - The incidence and mortality of prostate cancer is highly varied among populations and especially among blacks and whites. The incidence rates of all American populations have dramatically changed over the past 25 years. The recent increase in incidence has been attributed to prostate cancer screening. Although the incidence has increased over the past 25 years, the mortality rates although vastly different between populations have remained rather stable within populations. Prostate cancer is still a disease that primarily afflicts older men. The median age at diagnosis is 71 years for whites and 69 years for blacks. More than 80% are over the age of 65 years. Screening for prostate cancer has dramatically increased the number of men with local disease at diagnosis, but it is unclear whether screening and aggressive treatment have caused a decrease in mortality. PMID- 9858325 TI - The epidemiology of prostate cancer part II: the risk factors. AB - The epidemiology of prostate cancer gives us some clues that its etiology is likely both environmental and genetic. There is extreme country to country variability in prostate cancer mortality. Countries in which dietary fat intake is greater have been shown to have higher prostate cancer mortality rates leaving some to conclude that dietary fat causes prostate cancer. Migration studies show that men moving from Japan and China adopt increased risks of prostate cancer. Second- and third-generation Japanese Americans and Chinese Americans actually have risks of prostate cancer similar to white American men. This is highly suggestive that prostate cancer has an environmental influence. The differences in black-white mortality and newer data suggesting a higher mortality among Jamaican and Brazilian men of African descent suggest there may be a genetic predisposition to prostate cancer. Some have suggested certain polymorphisms increase prostate cancer risk, whereas others are searching for genetic mutations that may increase prostate cancer risk. Africans may have an increased prevalence of these genetic risk factors. Ultimately, the cause of prostate cancer is likely to be a combination of environmental and genetic factors. PMID- 9858326 TI - Hereditary prostate cancer in African-American families. AB - The recognition that family history is an important risk factor for the development of prostate cancer has led to the search for prostate cancer susceptibility genes. Although most prostate cancer linkage studies have focused on Caucasian families, there is epidemiological evidence indicating that the relative risk attributable to a positive family history is similar in African American and Caucasian-American families. There are data to support the existence of two potential prostate cancer susceptibility genes both on the long arm of chromosome 1; these have been called HPC1 and HPC2. Studies from us and others have shown that HPC1 may contribute to the clustering of prostate cancer in some African-American families. African-American families with multiple cases of prostate cancer should be encouraged to participate in genetic research studies with the goal of identifying the molecular basis for inherited prostate cancer susceptibility in this population. PMID- 9858327 TI - Race and serum prostate-specific antigen levels: current status and future directions. AB - Prostate cancer (PC) is a serious public health problem in the United States, especially in African-American men (AAs). AAs have higher incidence and mortality rates and present with higher stage and grade of PC than other racial/ethnic groups. Given the importance of serum prostate-specific antigen level (PSA) as a screening tool for PC and its prognostic value among patients with PC, exploring the relationship between PSA and race has gained enormous interest lately. The number of studies addressing such relationships has increased tremendously over the past few years. Most of these studies indicate a higher PSA in AAs than whites in men with or without PC even after adjustment for patients' age and prostate volume (in men without PC) and for PC grade and stage (in men with PC). The etiology of higher PSA levels in AAs is not completely understood and might be secondary to biological, environmental/socioeconomic reasons, or both. Studying serum PSA dynamics among AAs may help in elucidating the epidemiological features of PC in AAs. PMID- 9858328 TI - Pathology of premalignant lesions and carcinoma of the prostate in African American men. AB - The objective of this study was to summarize data describing the morphological characteristics of the spectrum of prostatic neoplasia in African-American men and compare them to their equivalents in Caucasian men. Studies that have emphasized the documentation of pathological features of prostate cancer or its putative precursors in African Americans are reviewed and are contrasted with comparable data concerning Caucasians. The author's experience is also presented. There is significant overlap between the pathological features of prostate cancer in African Americans and Caucasians in their clinical associations. The main pathological parameter reported with consistency to differ between the two groups is the observation that African-American patients present with a more advanced stage of the disease compared with Caucasian patients. Reports on differences in the grade and volume of the disease are less consistent. Comparative data on "preclinical," autopsy-type cancers and those concerning premalignant lesions indicate a similar prevalence of autopsy cancers between the two races while the prevalence and extent of high-grade prostatic intraepithelial neoplasia is higher in African-American men compared with Caucasian men. The difference is more evident in younger individuals. In conclusion, there are indications that African American men with clinically localized prostate cancer tend to have a more advanced pathological stage at the time of radical prostatectomy compared with Caucasian men. Our data also indicate a higher prevalence of a putative precursor for clinically evident prostate cancer, high-grade prostatic intraepithelial neoplasia in African-American men. The earlier onset and extensiveness of this lesion may contribute to the higher incidence of clinically evident prostate cancer in African Americans. PMID- 9858329 TI - Prostate cancer in the African American: is this a different disease? AB - Age-adjusted mortality rates from prostate cancer between 1991 and 1995 are approximately two times worse among African-American men than Caucasian men. Age specific mortality rates from prostate cancer are reported to be three times greater among African-American men compared with Caucasian men between the ages of 40 and 60 years. These statistics lead one to ask whether the difference in outcome is secondary to a more rapidly growing prostate cancer among African American men compared with Caucasian men, or is it a result of delayed diagnosis among African-American men, which would account for there being more advanced prostate cancer at presentation? This report demonstrates many histological and biological differences that may account for clinical outcome differences. PMID- 9858330 TI - The influence of race on the efficacy of curative radiation therapy for carcinoma of the prostate. AB - The purpose of this study was to examine the effect of race on the outcome of patients treated curatively with external beam irradiation for carcinoma of the prostate. The study was performed between January 1980 and December 1993 of 1,529 men with prostate cancer. Similar percentages of Caucasian men (CM) and African American men (AAM) had localized disease (stages T1 and T2) and advanced stage disease (stage T3). There was no difference in crude survival by race (P = .13). At 5 years, crude survival by race was 75% for CM and 73% for AAM. At 10 years, the crude survivals, were 50% and 40%, respectively. Disease-specific survival rates were equivalent for AAM and CM (P = .66). The 5-year disease-specific survival was 83% for CM and 85% for AAM. At 10 years, the disease-specific survival was 65% for CM and 69% for AAM. There was no difference in disease specific survival by race when stage-for-stage comparisons were made. Among those patients referred for curative radiation therapy, AAM and CM had a similar age, stage, and grade distribution. This study demonstrated that there was no difference in disease-specific survival between CM and AAM treated curatively with radiation for prostate cancer. PMID- 9858331 TI - Prostate cancer treatment outcome in blacks and whites: a summary of the literature. AB - The disparate mortality rates of prostate cancer among populations have led to the question "Is therapy as effective in blacks versus whites." A review of the cancer literature that has assessed black-white outcomes supports the conclusion that equal treatment yields equal outcomes among equal patients. Although epidemiological studies demonstrate that blacks are disproportionately diagnosed with higher stage and higher grade disease, clinical studies show that equal treatment yields equal outcome regardless of race. Patterns-of-care studies demonstrate that there is not equal treatment in the United States among black and white patients. PMID- 9858332 TI - Prostate cancer research and the National Cancer Institute. AB - The National Cancer Institute (NCI) is the largest funder of prostate cancer research in the United States and indeed the world. It is sponsoring a number of studies to answer the significant questions pertinent to prostate cancer and pertinent to black men. These include studies of epidemiology; cancer prevention, screening, and control; clinical treatment; and basic science. In addition, the NCI is charged with dissemination of research findings to physicians and the lay public. The following is a description of the NCI prostate cancer research portfolio including the scientific questions being addressed and how are they being addressed. PMID- 9858333 TI - Deconstructing (and reconstructing) cell migration. AB - An overriding objective in cell biology is to be able to relate properties of particular molecular components to cell behavioral functions and even physiology. In the "traditional" mode of molecular cell biology, this objective has been tackled on a molecule-by-molecule basis, and in the "future" mode sometimes termed "functional genomics," it might be attacked in a high-throughput, parallel manner. Regardless of the manner of approach, the relationship between molecular level properties and cell-level function is exceedingly difficult to elucidate because of the large number of relevant components involved, their high degree of interconnectedness, and the inescapable fact that they operate as physico chemical entities-according to the laws of kinetics and mechanics-in space and time within the cell. Cell migration is a prominent representative example of such a cell behavioral function that requires increased understanding for both scientific and technological advance. This article presents a framework, derived from an engineering perspective regarding complex systems, intended to aid in developing improved understanding of how properties of molecular components influence the function of cell migration. That is, cell population migration behavior can be deconstructed as follows: first in terms of a mathematical model comprising cell population parameters (random motility, chemotaxis/haptotaxis, and chemokinesis/haptokinesis coefficients), which in turn depend on characteristics of individual cell paths that can be analyzed in terms of a mathematical model comprising individual cell parameters (translocation speed, directional persistence time, chemotactic/haptotactic index), which in turn depend on cell-level physical processes underlying motility (membrane extension and retraction, cell/substratum adhesion, cell contractile force, front-vs.-rear asymmetry), which in turn depend on molecular-level properties of the plethora of components involved in governance and regulation of these processes. Hence, the influence of any molecular component on cell population migration can be understood by reconstructing these relationships from the molecular level to the physical process level to the individual cell path level to the cell population distribution level. This approach requires combining experimental, theoretical, and computational methodologies from molecular biology, biochemistry, biophysics, and bioengineering. PMID- 9858334 TI - Cell migration strategies in 3-D extracellular matrix: differences in morphology, cell matrix interactions, and integrin function. AB - Cell migration in extracellular matrix is a complex process of adhesion and deadhesion events combined with cellular strategies to overcome the biophysical resistance imposed by three-dimensionally interconnected matrix ligands. Using a 3-D collagen matrix migration model in combination with computer-assisted cell tracking for reconstruction of migration paths and confocal microscopy, we investigated molecular principles governing cell-matrix interactions and migration of different cell types. Highly invasive MV3 melanoma cells and fibroblasts are large and highly polarized cells migrating at low speed (0.1-0.5 microm/min) and at high directional persistence. MV3 melanoma cells utilize adhesive migration strategies as characterized by high beta1 integrin surface expression, beta1 integrin clustering at interactions with matrix fibers, and beta1 integrin-mediated adhesion for force generation and migration. In contrast, T lymphocytes and dendritic cells are highly mobile cells of lower beta1 integrin expression migrating at 10- to 40-fold higher velocities, and directionally unpredictable path profiles. This migration occurs in the absence of focal adhesions and largely independent of beta1 integrin-mediated adhesion. Whereas cell-matrix interactions of migrating tumor cells result in traction and reorientation of collagen fibers, partial matrix degradation, and pore formation, leukocytes form transient and short-lived interactions with the collagen lacking structural proteolysis and matrix remodeling. In conclusion, the 3-D extracellular matrix provides a spatially complex and biomechanically demanding substrate for cell migration, thereby differing from cell migration across planar ligands. Highly adhesive and integrin-dependent migration strategies detected in morphologically large and slowly migrating cells may result in reorganization of the extracellular matrix, whereas leukocytes favor largely integrin-independent, rapid, and flexible migration strategies lacking typical focal adhesions and structural matrix remodeling. PMID- 9858335 TI - Adhesion-guided in vitro morphogenesis in pure and mixed cell cultures. AB - The ability to understand and control the morphogenesis of mammalian cells is a fundamental objective of cell and developmental biology and tissue engineering research. Numerous processes, both biochemical and biophysical in nature, have been studied in an attempt to elucidate the mechanisms underlying this behavior. We focus here on the contributions of biophysical phenomena to the morphogenetic behavior of pure and mixed cell populations on solid surfaces in vitro. These principles are illustrated using characteristic liver tissue cells as a model system. The studies discussed demonstrate that cell-substratum and cell-cell adhesive forces are critical determinants of the ultimate morphology, cytoarchitecture, and organization achieved by these cells in vitro. PMID- 9858336 TI - Measuring actin dynamics in endothelial cells. AB - Cytoplasmic actin distributes between monomeric and filamentous phases in cells. As cells crawl, actin polymerizes near the plasma membrane of expanding peripheral cytoplasm and depolymerizes elsewhere. Thus, the finite actin filament lifetime, the diffusivity of actin monomer, and the distribution of actin between the polymer and monomer phases are key parameters in cell motility. The dynamics of cellular actin can be determined by following the evolution of fluorescence in the techniques of photoactivated fluorescence (PAF) or fluorescence recovery after photobleaching (FRAP) of microinjected actin derivatives. A mathematical model is discussed that measures monomer diffusion coefficients, filament turnover rates, and the fraction of actin polymerized from measurements of the evolution of fluorescence from a photoactivated band [Tardy et al. (1995) Biophys. J., 69:1674-1682; McGrath et al. (1998) Biophys. J., in press]. Applying this model to subconfluent endothelial cells shows that approximately 40% of the actin is polymer and that these filaments turn over on average every 6 minutes. This report discusses how PAF and FRAP can be combined with more traditional biochemistry to probe actin cytoskeleton remodeling in endothelial cells. PMID- 9858337 TI - Epidermal growth factor receptor-mediated motility in fibroblasts. AB - Cell motility is induced by many growth factors acting through cognate receptors with intrinsic tyrosine kinase activity (RPTK). However, most of the links between receptor activation and the biophysical processes of cell motility remain undeciphered. We have focused on the mechanisms by which the EGF receptor (EGFR) actuates fibroblast cell motility in an attempt to define this integrated process in one system. Our working model is that divergent, but interconnected pathways lead to the biophysical processes necessary for cell motility: cytoskeleton reorganization, membrane extension, formation of new adhesions to substratum, cell contraction, and release of adhesions at the rear. We postulate that for any given growth factor some of the pathways/processes will be actively signaled and rate-limiting, while others will be permissive due to background low-level activation. Certain couplings have been defined, such as PLCgamma and actin modifying proteins being involved in cytoskeletal reorganization and lamellipod extension and MEK being implicated in detachment from substratum. Others are suggested by complementary investigations in integrin-mediated motility, including rac in membrane protrusion, rho in new adhesions, myosin II motors in contraction, and calpain in detachment, but have yet to be placed in growth factor-induced motility. Our model postulates that many biochemical pathways will be shared between chemokinetic and haptokinetic motility but that select pathways will be activated only during RPTK-enhanced motility. PMID- 9858338 TI - Regulation of integrin-mediated adhesion during cell migration. AB - Migrating cells form dynamic and highly regulated adhesive interactions with their environment. In particular, integrin-mediated adhesions to the extracellular matrix (ECM) play a central role in cell migration. This review focuses on recent advances in understanding the adhesive mechanisms that regulate cell detachment at the rear of migrating fibroblasts and neutrophils. The contribution of several key adhesive regulators is discussed, including myosin mediated cell contractility, tyrosine phosphorylation, rho, calcium fluxes, and calpain. A challenge for future investigation will be to determine how adhesive events are spatially and temporally coordinated to promote productive directional cell movements. PMID- 9858339 TI - Signaling of de-adhesion in cellular regulation and motility. AB - Adhesion is a process that can be divided into three separate stages: (1) cell attachment, (2) cell spreading, and (3) the formation of focal adhesions and stress fibers. With each stage the adhesive strength of the cell increases. De adhesion can be defined as the process involving the transition of the cell from a strongly adherent state, characterized by focal adhesions and stress fibers, to a state of intermediate adherence, represented by a cell that is spread, but that lacks stress fibers terminating at adhesion plaques. We propose that this modification of the structural link between the actin cytoskeleton and the extracellular matrix results in a more malleable cellular state conducive for dynamic processes such as cytokinesis, mitogenesis, and motility. Anti-adhesive proteins, including thrombospondin, tenascin, and SPARC, rapidly signal de adhesion, potentially mediating proliferation and migration during development and wound healing. Intracellular signaling molecules involved in the regulation of de-adhesion are only beginning to be identified. Interestingly, many of the same signaling proteins recognized to play important roles during the process of adhesion have also been found to act during de-adhesion. Characterization of the precise mechanisms by which these signals modulate adhesive structures and the cytoskeleton will further our understanding of the regulation of adhesive strength and its function in cellular physiology. PMID- 9858340 TI - Chemoattractant-induced lamellipod extension. AB - The mammary adenocarcinoma cell line MTLn3 is chemotactic towards epidermal growth factor (EGF), and this induced motility is thought to promote breast cancer invasion and metastasis. Stimulation of MTLn3 cells with EGF results in the extension of a flat, thin structure filled with filamentous actin and termed a lamellipod. Lamellipod extension is dependent on actin polymerization and is localized to the border of adherent cells. The structure of EGF-stimulated lamellipods in MTLn3 cells is well suited to analysis of chemoattractant stimulated protrusion. Actin polymerization occurs within 200 nm of the extending edge of the lamellipod. Although extension of the lamellipod is not dependent upon interaction with the substratum, stabilization of the extended lamellipod is dependent on an adhesive substratum. Dorsal ruffling is suppressed during lamellipod extension. Tyrosine phosphorylation is reduced in preexisting focal contacts compared to new contacts induced by EGF stimulation. The coordination of turnover of focal contacts with lamellipod extension is proposed to result in polarized cell motility in response to gradients of chemoattractants. PMID- 9858341 TI - Contributions of the epidermal growth factor receptor to keratinocyte motility. AB - The epidermal growth factor (EGF) receptor plays a central role in numerous aspects of keratinocyte biology. In normal epidermis, the EGF receptor is important for autocrine growth of this renewing tissue, suppression of terminal differentiation, promotion of cell survival, and regulation of cell migration during epidermal morphogenesis and wound healing. In wounded skin, the EGF receptor is transiently up-regulated and is an important contributor to the proliferative and migratory aspects of wound reepithelialization. In keratinocytic carcinomas, aberrant expression or activation of the EGF receptor is common and has been proposed to play a role in tumor progression. Many cellular processes such as altered cell adhesion, expression of matrix degrading proteinases, and cell migration are common to keratinocytes during wound healing and in metastatic tumors. The EGF receptor is able to regulate each of these cellular functions and we propose that transient and dynamic elevation of EGF receptor during wound healing, or constitutive overexpression in tumors, provides an important contribution to the migratory and invasive potential of keratinocytes. PMID- 9858342 TI - Hepatocyte growth factor and Madin-Darby canine kidney cells: in vitro models of epithelial cell movement and morphogenesis. AB - It is becoming increasingly apparent that epithelial cell movement and changes in morphology are central to both development and regeneration of epithelial organs and are involved with pathological processes such as transformation of epithelia to carcinoma and metastasis. Hepatocyte growth factor (HGF) is a mesenchymally derived growth factor with pleiotrophic effects on epithelia depending on culture conditions. In vivo, HGF plays a role in mesenchymal-epithelial interactions. Madin-Darby canine kidney (MDCK) cells, which share many properties with polarized epithelia in vivo, are remarkably sensitive to HGF. In vitro models of HGF-treated MDCK cells have proven to be useful for the study of epithelial cell movement and changes in morphology. When cultured on plastic at low density, MDCK cells scatter in response to HGF. MDCK cells grown as cell suspensions in collagen gels form complex branching tubular structures in response to HGF. When cultivated as a monolayer on permeant supports, MDCK cells are well polarized with established E-cadherin mediated cell-cell junctions and dedifferentiate in response to HGF. Some of the mechanisms responsible for changes in cell movement and morphology that have been characterized using these models are summarized in this review. Models of MDCK cells exposed to HGF will continue to be useful in the study of epithelial cell movement and morphogenesis in vitro and will provide important clues into the cellular mechanisms important during in vivo epithelial processes such as organ development, regeneration, and transformation to carcinoma. PMID- 9858343 TI - Evaluation of the Gen-Probe PACE 2 assay for the detection of asymptomatic Chlamydia trachomatis and Neisseria gonorrhoeae infections in male arrestees. AB - BACKGROUND: The LET and Gen-Probe PACE 2 assay are used to screen male arrestees for evidence of infection with chlamydia and gonorrhea in the intake/booking area of the Jefferson Parish Correctional Center. GOAL OF THIS STUDY: To determine the accuracy of the Gen-Probe PACE 2 assay for the detection of asymptomatic infection with Chlamydia trachomatis and Neisseria gonorrhoeae infections in male arrestees. STUDY DESIGN: From December 1993 to March 1994, 196 arrestees younger than 22 years were screened with the Gen-Probe PACE 2 assay and McCoy shell vial culture for Chlamydia trachomatis. From April to October 1994, 444 arrestees of all ages were screened with the Gen-Probe PACE 2 assay and standard culture for Neisseria gonorrhoeae. The sensitivity, specificity, and predictive values of the Gen-Probe PACE 2 assay, compared with culture, were calculated with 95% confidence intervals. RESULTS: The Gen-Probe PACE 2 assay compared with culture had a sensitivity, specificity, and positive predictive value for Chlamydia trachomatis of 84%, 99%, and 93% and for Neisseria gonorrhoeae of 54%, 99.5%, and 78%. CONCLUSION: The Gen-Probe PACE 2 assay is useful for screening young males in this jail setting and is more accurate for detecting Chlamydia trachomatis compared with Neisseria gonorrhoeae. PMID- 9858344 TI - Failure of azithromycin therapy in gonorrhea and discorrelation with laboratory test parameters. AB - BACKGROUND: Azithromycin is efficacious in the treatment of chlamydial genital tract infection but less so in gonorrhea. However, MICs of azithromycin for gonococci from previously reported azithromycin treatment failures were consistently below the 'susceptible' MIC level of 2 mg/L. GOAL OF THIS STUDY: To examine gonococci not eliminated with 1 g azithromycin therapy to establish treatment outcome/MIC correlates in gonorrhea. STUDY DESIGN: The MICs and phenotypes of gonococci isolated from five cases of treatment failure after 1 g azithromycin therapy were determined and compared with the MICs of a systematic sample of routine isolates. RESULTS: Azithromycin MICs of gonococci from five cases of failed 1 g azithromycin treatment were 0.125 or 0.25 mg/L, well within the current 'susceptible' MIC range. None of the isolates were of the mtr phenotype. The MIC90 of a systematic sample of 219 gonococcal isolates was 0.25 mg/L. CONCLUSION: The antibiotic MIC/treatment outcome correlates that are usually found in gonorrhea do not apply for azithromycin. Current MIC criteria do not accurately define susceptibility or resistance of gonococci to azithromycin and by themselves do not predict the likely outcome of therapy. Pharmacokinetic factors may decrease the predictive value of MIC data. PMID- 9858345 TI - Development of a multi-item scale to quantitatively assess sexual behaviors and the transmission of high- and low-risk human papillomaviruses. AB - BACKGROUND: Epidemiologic research is frequently hindered by the inherent difficulty in quantifying the risk of sexually transmitted disease (STD) acquisition associated with individual patterns of sexual behavior. GOAL OF THE STUDY: To develop a quantitative sexual behavior risk scale and demonstrate its predictive validity in an assessment of risk factors for incident infection with human papillomaviruses (HPVs). STUDY DESIGN: Data from a prospective study of HPV infection in female university students was used to generate quantitative multi item sexual behavior scales which were used in Cox regression analyses. RESULTS: Although risk was incurred both in casual sexual encounters and in noncasual relationships, risk in these contexts were only weakly correlated. The construction of separate measurement scales was performed. CONCLUSIONS: Improved precision of measurement of STD risk demonstrated that significant risk was associated with different patterns of sexual behavior and was incurred in both casual and/or noncasual relationships. Scores on the sexual behavior risk scales were highly predictive of incident infection with HPV types of both high and low oncogenic potential. PMID- 9858346 TI - Trend in Chlamydia trachomatis infection among pregnant women in the past ten years in Japan: significance of Chlamydia trachomatis seroprevalence. AB - BACKGROUND AND OBJECTIVES: Chlamydia trachomatis infection is believed to be the most common bacterial sexually transmitted disease (STD) in industrialized countries. The objective of the current study was to assess the recent trend in the prevalence of C. trachomatis in Japan. GOAL OF THIS STUDY: To determine the trend in the seroprevalence for C. trachomatis among pregnant women in Nagasaki, Japan, during the past 10 years. STUDY DESIGN: The seroprevalence for C. trachomatis of 9,652 pregnant women of various ages screened in 1996 and 1997 was compared with those of 275 and 297 stocked samples from 1987 and 1992, respectively. Serum antibodies to C. trachomatis were detected by the enzyme immunoassay. Prospective samples of 33 seropositive cases were also analyzed to determine kinetics of the serum antibody titer. RESULTS: The seroprevalence has decreased in all age groups during the last 10 years. More than 70% of seropositive cases converted to be seronegative within 10 years. CONCLUSION: The prevalence of C. trachomatis has been decreasing among Japanese pregnant women. PMID- 9858347 TI - Quinolone-resistant Neisseria gonorrhoeae: the beginning of the end? Report of quinolone-resistant isolates and surveillance in the southwestern United States, 1989 to 1997. AB - BACKGROUND: Fluoroquinolones are one of the most widely used treatments for gonorrhoeae. Changes in the susceptibility of Neisseria gonorrhoeae to these agents may threaten their use. GOAL OF THIS STUDY: To report several resistant strains (>1 mcg/ml) isolated in the western United States and to evaluate the prevalence of strains with reduced susceptibility (ofloxacin 0.25 mcg/ml, ciprofloxacin 0.06 mcg/ml). STUDY DESIGN: The microbiology and epidemiology of three resistant strains were characterized and 12,761 other strains were evaluated for fluoroquinolone susceptibility as part of the Gonococcal Isolate Surveillance Project of the Centers for Disease Control and Prevention. RESULTS: Fluoroquinolone-resistant strains may appear sporadically. The prevalence of isolates with reduced susceptibility to fluoroquinolones remains low in the Southwest region of the United States. CONCLUSIONS: Continued active surveillance is needed to detect and control the spread of quinolone-resistant Neisseria gonorrhoeae. PMID- 9858348 TI - Hepatitis C virus infection in Chicago women with or at risk for HIV infection: evidence for sexual transmission. AB - BACKGROUND AND OBJECTIVES: The importance of sexual transmission of hepatitis C virus (HCV) infection is unclear. We attempted to define its role in women with or at risk for HIV infection. GOAL OF THIS STUDY: To ascertain if high-risk sexual behavior was independently associated with HCV infection. STUDY DESIGN: Risk factors were assessed cross-sectionally in Chicago women newly enrolled in the Women's Interagency HIV Study. Women who had (n = 243) or were at risk for HIV infection (n = 53) were tested for HCV antibodies (Ab). RESULTS: Of 296 women, 123 (42%) were HCV Ab positive; prevalence was 90% in women who injected drugs (IDU) compared with 12% in noninjectors (odds ratio [OR], 64.0, 95% confidence interval [CI], 29.9 to 137.0). A multivariate model showed associations with IDU (OR, 110.3, 95% CI, 33.3 to 365.8), prior gonorrhea (OR, 3.6, 95% CI, 1.4 to 8.9), and sex with a male IDU (OR, 2.7, 95% CI, 1.1 to 7.0). CONCLUSION: Injection drug use is the strongest predictor of HCV infection, but sexual risk factors are also independently associated. PMID- 9858349 TI - Detection of Neisseria gonorrhoeae infection by ligase chain reaction testing of urine among adolescent women with and without Chlamydia trachomatis infection. AB - BACKGROUND AND OBJECTIVES: Culture, the conventional method for detection of Neisseria gonorrhoeae, requires invasive sampling and stringent specimen transport conditions. The recently developed ligase chain reaction test (LCR; Abbott Laboratories; North Chicago, IL) allows noninvasive sampling and stable transport conditions, but has not been evaluated with specimens from adolescent populations. GOAL OF THIS STUDY: To perform a comparative evaluation of a commercial LCR test and culture for the diagnosis of N. gonorrhoeae in adolescent women. STUDY DESIGN: Urine and endocervical swab specimens from 330 teenage women seen in two public health adolescent clinics were tested by LCR and culture. For resolution of discordant results, a polymerase chain reaction (PCR) test was developed that directly amplifies N. gonorrhoeae DNA from urine samples processed for LCR. RESULTS: Thirty-one of 330 (9.4%) cervical specimens were culture positive for N. gonorrhoeae, and 30 of 330 (9.1%) urine specimens were positive by LCR. After resolution of 13 discordant results, the sensitivity, specificity, and positive and negative predictive values of LCR for urine were 88.2%, 100%, 100%, 98.7%, respectively, and for culture of endocervical specimens were 82.3%, 98.9%, 90.3% and 98%, respectively. CONCLUSIONS: Although more expensive than culture, LCR offers a sensitive means for the detection of N. gonorrhoeae in urine samples and may be useful for this purpose in settings where pelvic examinations are difficult to perform and simultaneous detection of N. gonorrhoeae and Chlamydia trachomatis is advantageous. PMID- 9858350 TI - Risk assessment to improve targeting of HIV counseling and testing services for STD clinic patients. AB - BACKGROUND AND OBJECTIVE: To determine whether self-administered risk assessment could improve targeting of HIV counseling and testing in an STD clinic. STUDY DESIGN: Computerized records from the Prince George's County, Maryland, STD clinic from 1993 through 1996 were used to develop and test models for predicting a positive HIV test. In 1996, a self-administered risk assessment was compared with a counselor's risk assessment of the same patient. RESULTS: Testing the 10% of patients at highest risk would identify 39% of those who were HIV-positive; testing 70% of the patients could identify 92% of those who were HIV-positive. In 1996, 2,288 patients completed the self-administered HIV risk assessment. The same number of HIV-positive persons (7 [28%]) were identified using either self assessment or face-to-face interview. CONCLUSIONS: Selectively offering voluntary HIV testing based on risk assessment would not be useful because it would miss many infected persons. If prevention counseling cannot be offered to everyone, it could be targeted to those who report a risk by self-assessment. PMID- 9858351 TI - A geographic relation between alcohol availability and gonorrhea rates. AB - BACKGROUND AND OBJECTIVES: The availability of alcohol measured as alcohol outlet density is associated with numerous alcohol-related outcomes in small area analysis. A number of studies suggest that high-risk sexual behavior should also be considered an alcohol-related outcome. GOAL OF THIS STUDY: To assess the geographic relationship between alcohol availability and high-risk sexual behavior at the neighborhood level. STUDY DESIGN: Ecological analysis of the geographic relation between off-premise, on-premise, and total alcohol outlet density and reported gonorrhea rates among 155 urban residential census tracts in New Orleans during 1995. RESULTS: All alcohol outlet density variables were positively related to gonorrhea rates. Off-premise outlets per square mile was most strongly related to gonorrhea rates (beta +/- SE) (beta = 0.582+/-0.073), accounting for 29% of the variance in gonorrhea rates. Interpreted as an elasticity, a 10% increase in off-sale alcohol outlet density accounts for a 5.8% increase in gonorrhea rates. Including the covariates percent black and percent unemployed to the model reduced but did not remove the effect of off-sale outlet density (beta = 0.192+/-0.047). CONCLUSIONS: These results indicate there is a geographic relationship between alcohol outlet density and gonorrhea rates at the census tract level. Although these results cannot be interpreted causally, they do justify a public health intervention as a next step in defining the relation between alcohol availability and high-risk sexual behavior. PMID- 9858352 TI - Treponemal specific tests for the serodiagnosis of syphilis. Syphilis and HIV Study Group. AB - OBJECTIVES: To determine the rate of concordance of the Microhemagglutination Assay for Antibodies to T. pallidum (MHA-TP) and the Fluorescent Treponemal Antibody-Absorption test (FTA-ABS) prior to therapy in patients with early stage syphilis and to assess the incidence of and associated risk factors for seroreversion of these treponemal specific tests during the first year after therapy for early syphilis. DESIGN: Multicenter, prospective, cohort treatment study of patients with early syphilis. METHODS: Five hundred twenty-five patients were enrolled in a study to evaluate the response of early syphilis to either benzathine penicillin 2.4 million units intramuscularly once or this therapy plus amoxicillin 2 g and probenecid 500 mg orally both three times daily for 10 days. Serologic and clinical follow-up was conducted at intervals over 1 year. MHA-TP and FTA-ABS tests were performed on serologic specimens from each patient visit. RESULTS: Enrollment specimens showed 5% discordant MHA-TP and FTA-ABS results with 85% of these demonstrating a nonreactive MHA-TP. This occurred most commonly in primary syphilis. In patients who had a 1-year serologic follow-up with FTA ABS or MHA-TP, seroreversion occurred in 9% and 5% of cases, respectively. No association between HIV-seropositivity and TST seroreversion was demonstrated. CONCLUSION: The MHA-TP may be less sensitive than the FTA-ABS for identifying patients with primary syphilis. Treponemal specific tests may become nonreactive during the first year after therapy for early syphilis. PMID- 9858353 TI - Group counseling to prevent sexually transmitted disease and HIV: a randomized controlled trial. AB - OBJECTIVE: To compare prevention effectiveness of multisession group counseling with standard HIV prevention counseling for reducing risk behaviors and new sexually transmitted diseases (STDs). METHODS: Small groups of consenting STD clinic patients were randomized to four 1-hour small group counseling interventions based on the information-motivation-behavioral skills (IMB) model with a booster session at 2 months or to the standard two 20-minute individual counseling sessions. Follow-up interviews and examinations were 2, 6, 9, and 12 months after enrollment. RESULTS: From March 1992 through June 1993, 996 (59%) of 1,681 eligible persons were enrolled; 32 (3%) tested HIV-positive and were excluded. Intervention attendance was 98% for one session, and 47% attended four or five counseling sessions. Follow-up was similar for both groups: 72% attended at least once; 47% returned at 12 months. Both groups had similar increases in condom use and decreases in number of partners, and similar number of new infections with gonorrhea (14%), chlamydia (10%), or syphilis (2%). CONCLUSIONS: Two 20-minute counseling sessions were as effective as four 1-hour group sessions for reducing risk behavior and STD incidence in an STD clinic patient population. PMID- 9858354 TI - Partner notification in the real world: a four site time-allocation study. AB - BACKGROUND AND OBJECTIVES: Although partner notification has been a long-standing intervention and prevention strategy for sexually transmitted diseases (STD), variations in partner notification practices across sites have never been documented. GOALS OF THE STUDY: To describe provider-assisted partner notification practices in four STD programs in the United States. STUDY DESIGN: Eleven disease intervention specialists (DIS) in each of three urban sites and seven DIS in one rural site documented their activities and clients for 14 working days using a personal digital assistant. RESULTS: Of 2,506 recorded activity hours across sites, 37.4% of the recorded time was spent on partner notification (PN) activities with 1148 clients. Field visits to locate contacts accounted for the largest proportion of time spent on PN. Overall, PN clients were cases of or were contacts to nonprimary and secondary (P&S) syphilis (39.6%), gonorrhea (25.5%), chlamydia (18.0%), HIV/AIDS (10.4%), and P&S syphilis (6.5%). CONCLUSION: The activities which constitute PN, the diseases for which PN is used, and the time spent on each PN client vary across sites. More research is needed on the determinants of these variations and their association with the ultimate goal of disease prevention. PMID- 9858355 TI - The association of false-positive rapid plasma reagin results and HIV infection. AB - BACKGROUND AND OBJECTIVES: To study the relationship between a false positive rapid plasma reagin (RPR) result (FP), syphilis, and HIV infection in our patients. METHODS: A prospective study of the incidence of FP tests and syphilis in the general population and its relationship to HIV infection over a period of 6 months. RESULTS: 8.76% of the population were HIV positive. False positives were found in 15% and 1.2% of the HIV infected and noninfected patients, respectively; the attributable risk for HIV was 14.97. Syphilis was found in 5% and 0.9% of the positive and negative HIV patients, respectively; the attributable risk for HIV was 5.4. CONCLUSIONS: The incidence of false positive RPR results in the HIV-infected population is significantly higher than that of the non-HIV-infected patients. PMID- 9858356 TI - The saga of the nerve growth factor. PMID- 9858357 TI - Endurance training increases acetylcholine receptor quantity at neuromuscular junctions of adult rat skeletal muscle. AB - The aim of the study was to test the hypothesis that a 16 week endurance training program would alter the abundance of endplate-associated nicotinic acetylcholine receptors (nAChR) in various rat skeletal muscles. We found a 20% increase in endplate-specific [125I]alpha-bungarotoxin binding in several muscles of trained rats, accompanied by equal susceptibility of toxin binding to the inhibitory effect of D-tubocurarine in sedentary and trained muscles. We conclude that the neuromuscular junction adaptations that occur with increased chronic activation include an increase in nAChR number. Results of experiments designed to determine nAChR turnover also suggest that this effect is mediated by an alteration in the receptor's metabolic state. The potential implications and mechanisms of this adaptation are discussed. PMID- 9858358 TI - 6-OHDA denervation substantially decreases DCC mRNA levels in rat substantia nigra compacta. AB - The function of deleted in colorectal cancer (DCC) protein, a member of the immunoglobulin superfamily of cell adhesion molecules, in the adult CNS is unknown. Recently the transcript encoding DCC has been shown to be expressed in a variety of rat brain regions, including the substantia nigra pars compacta and the striatum, which encompasses the nigrostriatal dopaminergic system. In the present study DCC mRNA expression in substantia nigra, striatum, dentate gyrus and piriform cortex was investigated in adult rats using in situ hybridization histochemistry following unilateral injections of 6-hydroxydopamine (6-OHDA) in the median forebrain bundle. DCC mRNA levels were greatly reduced in the substantia nigra ipsilateral to the 6-OHDA lesion compared to those on the contralateral side while there was no apparent effect on DCC mRNA levels in the other regions analysed. These data indicate expression of DCC mRNA in dopamine neurones of the substantia nigra pars compacta and support a role for DCC in the adult CNS, with potential involvement in the function of central dopamine neurones. PMID- 9858359 TI - The time course of auditory perceptual learning: neurophysiological changes during speech-sound training. AB - Here we report that training-associated changes in neural activity can precede behavioral learning. This finding suggests that speech-sound learning occurs at a pre-attentive level which can be measured neurophysiologically (in the absence of a behavioral response) to assess the efficacy of training. Children with biologically based perceptual learning deficits as well as people who wear cochlear implants or hearing aids undergo various forms of auditory training. The effectiveness of auditory training can be difficult to assess using behavioral methods because these populations are communicatively impaired and may have attention and/or cognitive deficits. Based on our findings, if neurophysiological changes are seen during auditory training, then the training method is effectively altering the neural representation of the speech/sounds and changes in behavior are likely to follow. PMID- 9858360 TI - Aggressive behaviour in transgenic mice expressing APP is alleviated by serotonergic drugs. AB - Transgenic mouse strains were generated that overexpress human APP or clinical mutants of APP. All transgenic mouse strains that over-express APP displayed essentially the same phenotype of disturbed behaviour, differential glutamatergic responses, deficits in maintenance of long-term potentiation and premature death, but formation of amyloid plaques was seen in the highest expressing APP/London transgenic mice only. Apart from cognitive deficits, the APP transgenic mice were characterized by aggressive behaviour, which was pharmacologically alleviated with 8-OH-DPAT and buspirone, two serotonergic agonists. The atypical neuroleptic drug risperidone was equally active in this regard. The data establish an important aspect of the transgenic mice as experimental models for behavioural aspects of Alzheimer's disease, in addition to other early and late symptoms. PMID- 9858361 TI - Effect of inhibition of glucose and fat metabolism on galanin-R1 receptor mRNA levels in the rat hypothalamic paraventricular and supraoptic nuclei. AB - Using in situ hybridization histochemistry we have studied the effect of glucose and fat deprivation on galanin-R1 receptor (GAL-R1-R) mRNA levels in the rat paraventricular (PVN) and supraoptic (SON) nuclei after single and repeated i.p. administration of a glucose antimetabolite 2-deoxy-D-glucose (DG), as well as of a fatty acid antimetabolite, sodium mercaptoacetate (MA), treatments known to increase food intake. Both DG and MA injections caused an increase in levels of GAL-R1 mRNA transcripts in the PVN and SON. These results indicate that glucose and fat deprivation increase the sensitivity of PVN and SON neurons to galanin, and that regulation of receptor levels may be an important mechanism in galaninergic signalling. PMID- 9858362 TI - Alteration of digital representations in somatosensory cortex in focal hand dystonia. AB - Focal hand dystonia involves a loss of motor control of one or more digits; it is associated with the repetitive, synchronous movements of the digits made by musicians over periods of many years. Magnetic source imaging revealed that there is a smaller distance (fusion) between the representations of the digits in somatosensory cortex for the affected hand of dystonic musicians than for the hands of non-musician control subjects. The data suggest that use-dependent susceptibility to digital representation fusion in cortex may be involved in the etiology of focal dystonia. A successful therapy for the condition has been developed based on this consideration. PMID- 9858363 TI - Identification of a serine protease with nerve growth promoting activity from snake venom. AB - A new protein with nerve growth promoting activity was purified from the crude venom of the Agkistrodon halys Pallas, a Chinese snake. Its amino-terminal sequence unexpectedly showed high homology with serine proteases, suggesting that it is a new member of the serine protease family. It also cross-reacted with antibodies against thrombin-like enzyme and possessed weak arginine esterase activity, amounting to about 3% of the activity of trypsin. However, its nerve growth promoting activity was comparable to that of nerve growth factor (NGF). It was named NGF-like protease (NLP). Northern blot analysis further demonstrated different patterns of induction of c-myc, vgf and trkA mRNA transcription in PC12 pheochromocytoma cells treated with NGF and NLP, respectively. These data suggested that NLP represents a novel potent neurotrophic factor. PMID- 9858364 TI - Dystrophins in neurohypophysial lobe of normal and dehydrated rats: immunolocalization and biochemical characterization. AB - Dystrophin, utrophin and dystroglycan are present not only in muscle but also in brain. In muscle, they link the extracellular matrix to the cytoskeleton. Their function in brain is not understood. Here we show their presence in the hypothalamo-neurohypophysial system which secretes the neurohormones oxytocin and vasopressin. Using immunocytochemistry, we showed that dystrophins are present in the neurohypophysis of control rats. After water deprivation, immunoreactivity dramatically decreased and appeared in axonal swellings in the hypothalamic tract. Dystrophin immunostaining can be ascribed to dystrophin and/or utrophin as well as the DMD (Duchenne Muscular Dystrophy) gene short products Dp140 and Dp71 as revealed by Western immunoblots of synaptosomes isolated from neurohypophyses of control rats. In synaptosomes isolated from rats under water deprivation, the immunoreactivity entirely disappeared. Further biochemichal characterization of isolated neurosecretory granules (NSG) showed that Dp140 and Dp71 are enriched in the NSG stored in the swellings of the neurohypophysis whereas the NSG of the nerve endings are devoid of these proteins. In addition we observed that the presence of beta-dystroglycan and actin correlates with the presence of dystrophins. Our data favor a direct implication of the dystrophins and/or utrophin, dystroglycan and actin in the neurosecretory processes of the hypothalamo-neurohypophysial system. PMID- 9858365 TI - Reproductive experience reduces striatal dopaminergic responses in freely moving female rats. AB - Reproductive experience influences basal and pregnancy profiles of circulating prolactin levels in women and female rats, respectively. Endocrine responses to dopaminergic antagonists are modified by reproductive experience as well. Striatal extracellular dopamine metabolites were measured in vivo by HPLC-ED in perfusates obtained by microdialysis in non-anaesthetized, freely moving, intact and ovariectomized, nulliparous and primiparous rats. Data were collected for at least 7 h. This period always included the light-dark shift at 18:00 h. In a second experiment, microdialysis was performed in ovariectomized nulliparous and primiparous rats treated with haloperidol (1.0 mg/kg s.c.). During the 1 h before and after the onset of the light-dark shift, HVA concentrations in the perfusates increased in nulliparous compared with primiparous rats. The haloperidol-induced increase in DOPAC and HVA was less intense in primiparous rats than that in nulliparous ovariectomized rats. These data revealed a different pattern of striatal dopaminergic anticipation and response to the shift in the light-dark cycle in nulliparous as compared with primiparous intact females. In addition, a distinct striatal dopaminergic response to haloperidol was observed in primiparous as compared to nulliparous ovariectomized rats. The results suggest that reproductive experience can modulate the activity of dopaminergic terminals in the striatum. PMID- 9858366 TI - Short CAG repeats within the hSKCa3 gene associated with schizophrenia: results of a family-based study. AB - In a family-based association study we investigated transmission of a multiallelic CAG repeat in a novel neuronal potassium channel gene, hSKCa3, in 59 parent/ offspring trios. In contrast to recent reports of an association of moderately large repeats with schizophrenia in case-control studies, our findings indicate that short CAG repeats (< or=19 repeats) are transmitted at an increased frequency to schizophrenic offspring (p=0.014), particularly among familial cases (p=0.007). No evidence for a parent-of-origin effect was found. Multiallelic TDT procedure showed no association of individual CAG repeats to schizophrenia. Further studies using family-based designs should clarify whether hSKCa3 is a susceptibility factor to schizophrenia or co-segregates with a major disease gene in tight linkage. PMID- 9858367 TI - Modulation of the neurotransmitter release in rat cerebellar neurons by GRO beta. AB - We report here that, in cultured cerebellar granule cells, the CXC chemokine GRObeta stimulates the signaling pathway of the extracellular signal-regulated kinases, and enhances both evoked and spontaneous postsynaptic currents in patch clamped Purkinje neurons from rat cerebellar slices. The GRObeta-induced enhancement of the excitatory post-synaptic currents evoked by stimulating the parallel fibres is blocked by the inhibitor of the extracellular signal-regulated kinases pathway PD98059, which also reduces both basal frequency of spontaneous post-synaptic currents and mean amplitude of evoked excitatory post-synaptic currents. Our results suggest that GRObeta modulates neurotransmitter release in the cerebellum through the activation of the extracellular signal-regulated kinases pathway. PMID- 9858368 TI - Intact recognition of vocal expressions of fear following bilateral lesions of the human amygdala. AB - A recent case study found that bilateral damage to the amygdala impairs the normal appraisal of vocal expressions of fear. However, the single source of evidence for this auditory emotion-processing impairment is from a patient with extra-amygdaloid damage that may include the basal ganglia, which have been shown to be important for prosody evaluation. In this study we provide evidence of preserved evaluation of vocal expressions of fear in a female patient (S.P.) with bilateral damage to the amygdala but with intact basal ganglia. This same patient has previously been shown to be impaired in the evaluation of facial expressions, including fear. These results indicate that the analysis of nonverbal signals of fear from different input channels are dissociable, being at least partially dependent on different brain structures. We suggest that the amygdala, in conjunction with the basal ganglia, may support the normal appraisal of auditory signals of danger. PMID- 9858369 TI - Early assessment of motor dysfunctions aids in successful occlusion of the middle cerebral artery. AB - Occlusion of the rodent middle cerebral artery by embolism, using an intraluminal filament, produces behavioral alterations which resemble many symptoms associated with stroke. This model has been used to examine treatment interventions for the disease, however, variable success rate in completely blocking the middle cerebral artery may present inconclusive interpretation of the data. To detect successful occlusion of the middle cerebral artery, we demonstrate here sensitive and reliable behavioral parameters including the elevated body swing test, the postural tail-hang test, the spontaneous rotational test, and the forelimb akinesia test. These assays provide a criterion for identifying animals with incomplete occlusion which could promote host-related spontaneous recovery and might confound true effects of experimental therapies on ischemia-induced dysfunctions. From a practical standpoint, the early reliable identification of partial cerebral ischemia aids in immediate and efficient adjustments of the surgical procedure to create a complete and stable ischemia stroke animal model. PMID- 9858370 TI - The role of the ventromedial nucleus of the hypothalamus in epileptogenesis. AB - Flurothyl kindling initiates a time-dependent process that results in a facilitated propagation from the forebrain to the brainstem seizure system and in an increase in the complexity of behavioral seizure expression. We investigated the involvement of the ventromedial nucleus of the hypothalamus (VMH) in mediating this facilitated propagation between these seizure systems. Bilateral ibotenic acid lesions of the VMH, but not the dorsomedial nucleus of the hypothalamus (DMH), resulted in a disruption in the propagation of seizure activity from the forebrain to the brainstem. Moreover, VMH lesioned mice were able to express brainstem seizures following minimal corneal electroconvulsive shock (mECS). Together, our results indicate that the VMH is a critical substrate involved in propagating seizure activity between the forebrain and brainstem, but is not involved in the expression systems necessary for forebrain or brainstem seizure manifestations. PMID- 9858371 TI - Cerebral control of unimanual and bimanual movements: an H2(15)O PET study. AB - We measured regional cerebral blood flow (rCBF) during unimanual and bimanual movements using H2(15)O PET. Six healthy volunteers performed unimanual, bimanual symmetric and bimanual-asymmetric ballistic finger movements. The study was designed to minimize anticipation and preparation of movements. Data were analysed using SPM. Unpredictably paced unimanual movements resulted in significant activation of contralateral primary motor-somatosensory cortex (M1 S1) and mesial frontal cortex (p < 0.001). Performance of symmetric bimanual movements resulted in bilateral activation, but no additional activation of mesial frontal cortex was shown. Comparison of asymmetric with symmetric bimanual movements revealed additional recruitment of mesial frontal cortex (p< 0.001). We suggest that rostral mesial frontal cortex facilitates asymmetric non-mirrored bimanual finger movements. PMID- 9858372 TI - Expression of extraocular-superfast-myosin heavy chain in rat laryngeal muscles. AB - Superfast myosin heavy chain (MHC), which is found in jaw-closing muscle and extraocular muscle (EOM), may also be found in rat laryngeal muscles. Immunostaining and Western blot using anti-EOM antibody were performed to identify and localize EOM MHC in laryngeal muscles. Specific reactivity of laryngeal IIL MHC was confirmed by Western blot and on immunostaining, all fibers in the lateral part of thyroarytenoid muscle reacted with EOM antibody. A scattered pattern of positive fibers was observed in the medial part of the thyroarytenoid, the posterior cricoarytenoid and the lateral cricoarytenoid muscles. EOM MHC was not detected in the cricothyroid muscle. The expression of EOM MHC in rat laryngeal muscle is consistent with the functional demands of the airway protection reflex. PMID- 9858373 TI - Nitric oxide synthase in the auditory brain stem. AB - The auditory superior olivary complex (SOC), a group of inter-related brain stem nuclei, sends efferents to a variety of neuronal structures including the cochlea and the inferior colliculus. The present study conducted in rats and hamsters sought to investigate whether the neuroactive substance nitric oxide (NO) is produced in the rodent SOC. Immunohisto- and cytochemistry of the NO-synthesizing enzyme neuronal NO-synthase (nNOS), and histochemistry (NADPH-diaphorase activity of nNOS) was used. Perikarya stained by either method were found in the medial nucleus of the trapezoid body and in periolivary nuclei projecting to the cochlea, while terminals were observed mainly in the lateral superior olivary nucleus and in the superior paraolivary nucleus. These data demonstrate the existence of nNOS in the rodent SOC and provide evidence that nitric oxide acts as neuroactive substance in the auditory brain stem and the organ of Corti. PMID- 9858374 TI - D-cycloserine, a partial NMDA receptor-associated glycine-B site agonist, enhances reversal learning, but a cholinesterase inhibitor and nicotine has no effect. AB - The present study examined the efficacy of single and combined treatments with an anticholinesterase, tetrahydroaminoacridine, nicotine and a glycine-B site partial agonist, D-cycloserine, in alleviating the water maze reversal learning defect induced by a medial septal lesion. D-cycloserine (3 and 10 mg/kg) improved reversal learning. Tetrahydroaminoacridine (1 and 3 mg/kg) and nicotine (0.1 and 0.3 mg/kg) had no effect on reversal learning. A combination of tetrahydroaminoacridine 3 mg/kg or nicotine 0.3 mg/kg and D-cycloserine 10 mg/kg was not more effective than D-cycloserine 10 mg/kg alone in improving reversal learning. This suggests that stimulation of NMDA mechanisms may more effectively improve in medial septal-lesioned rats reversal learning processes than stimulation of cholinergic activity. PMID- 9858375 TI - Neuronal activity in the cholinoceptive basal forebrain of freely moving narcoleptic dobermans. AB - Cholinergic stimulation in the basal forebrain (BF) triggers cataplexy in canine narcolepsy. Extracellular single unit recordings in the BF were carried out in freely moving narcoleptic dogs to study the neuronal mechanisms mediating cataplexy induction in the BF. Among the 64 recorded neurons, 12 were wake active, three were slow wave sleep (SWS)-active, 17 were wake-/REM-active, 11 were REM sleep-active, three were cataplexy-active, and the other 18 were state independent. Systemic administration of physostigmine, a cholinesterase inhibitor, induces status cataplecticus, decreases SWS and increases acetylcholine levels in the BF. Firing of most of the state-dependent neurons in the BF was significantly modified by physostigmine. Some of these neurons may thus mediate sleep stage changes or the effect on cataplexy observed after cholinergic stimulation in the BF. PMID- 9858376 TI - Slow intrinsic optical signals in the rat spinal dorsal horn in slice. AB - Tetanic stimulation of high-threshold primary afferent fibers in the dorsal root was found to elicit intrinsic optical signals (IOSs) in transverse slices of 11- to 20-day-old rat spinal cords. The IOS, lasting for 30 s or longer, was most prominent in the lamina II of the dorsal horn. Treatment with a Na+-K+-2Cl- co transport blocker, furosemide, abolished the IOS, suggesting that the origin of the IOS is the cellular swelling due to an activity-dependent rise in extracellular K+. Substance P antagonist spantide, glutamate antagonists 2-amino 5-phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione, and the mu opioid agonist [d-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin suppressed IOSs. Thus, IOSs represent at least in part the slow excitatory response that is known to be generated in dorsal horn neurons after tetanic activation of unmyelinated afferent fibers. PMID- 9858377 TI - Human primary visual cortex and lateral geniculate nucleus activation during visual imagery. AB - The functional magnetic resonance (fMRI) technique can be robustly used to map functional activation of the visual pathway including the primary visual cortex (V1), the lateral geniculate nucleus (LGN), and other nuclei of humans during visual perception stimulation. One of the major controversies in visual neuroscience is whether lower-order visual areas involve the visual imagery process. This issue was examined using fMRI at high magnetic field. It was demonstrated for the first time that the LGN was activated during visual imagery process in the human brain together with V1 and other activation. There was a tight coupling of the activation between V1 and the LGN during visual imagery. PMID- 9858378 TI - Novel alternative splicing in the 5' exon of the neurotrophin-3 gene. AB - The neurotrophin-3 (NT-3) gene has previously been reported to consist of three exons including two 5' short untranslated exons and a 3' long exon encoding the entire protein, and to give rise to two classes of transcripts by alternative splicing of the 5' exons to the 3' coding exon. In the present study, we demonstrated the presence of at least four new classes of transcripts of the NT-3 gene, in addition to the two known transcripts. The present finding proposes the further complexity of the regulational mechanism for NT-3 expression. PMID- 9858379 TI - Relationship of specific EEG frequencies at specific brain areas with performance. AB - This study shows that incorrect responses are preceded by different EEG characteristics than correct responses, and that these differences appear in specific brain regions that participate in each particular task. EEGs were recorded in children during three different tasks: color discrimination (CDT), verbal working memory (VWM), and word categorization task (WCT). EEG segments previous to the presentation of the stimulus were analysed. Incorrect responses were preceded by lower EEG power values at 7.8 Hz in posterior temporal and right parietal leads in CDT, 8.59 and 9.36 Hz in frontal areas in VWM, and 10.72 Hz in the left hemisphere in WCT. In the former task > 1.56 Hz power in frontal areas prior to an incorrect response was also observed. PMID- 9858380 TI - LTP in CA1 of the adult rat hippocampus and voltage-activated calcium channels. AB - It is difficult to induce long-term potentiation (LTP) in CA1 of hippocampal slices from 120-day-old rats when a single 100 Hz, 1 s tetanus is administered in extracellular solution containing 2.0 mM calcium and 2.0 mM magnesium. However, in the presence of 2.5 mM calcium and 1.3 mM magnesium LTP is reliably induced by this same stimulus. Although the amplitude of LTP is similar to that observed in slices from 30-day-old rats, LTP in slices from mature rats is not inhibited by MK-801 but is blocked by nifedipine. These results suggest that factors contributing to LTP in slices from mature rats require careful consideration under different experimental paradigms. PMID- 9858381 TI - [3H]acetylcholine release from rat amacrine-like neurons is inhibited by adenosine A1 receptor activation. AB - We studied the effect of endogenous adenosine on the release of [3H]acetylcholine ([3H]ACh) in cultures enriched (96.4+/-0.4%) in rat cholinergic amacrine-like neurons, as determined by labeling with an antibody against choline acetyltransferase. A small population of these cells also contained GABA. Using these cultures we observed that both [3H]ACh release, which was largely Ca2+ dependent, and 45Ca2+ influx, evoked by depolarization with 50 mM KCl, were increased when adenosine A1 receptor activation was prevented by removal of endogenous adenosine with adenosine deaminase, or by application of the A1 receptor antagonist DPCPX. Our results indicate that, in cultured rat amacrine like neurons, the activation of A1 receptors decreases calcium influx and, thereby, inhibits [3H]ACh release. PMID- 9858383 TI - Cerebral ischemia and CNS transplantation: differential effects of grafted fetal rat striatal cells and human neurons derived from a clonal cell line. AB - Stroke mortality has declined over recent decades, prompting a demand for the development of effective rehabilitative therapies for stroke survivors. This effort has been facilitated by significant progress in replicating the behavioral and neuropathological changes of authentic human cerebral ischemia using relevant animal models. Since the rodent model of middle cerebral artery occlusion mimics several motor abnormalities seen in clinical cerebral ischemia, we have utilized this model to investigate treatment strategies for stroke. The present study explored the potential benefits of neural transplantation of fetal rat striatal cells or human neurons derived from a clonal embryonal carcinoma cell line to correct the abnormalities associated with cerebral ischemia. We report here that ischemia-induced behavioral dysfunctions were ameliorated by the neural grafts as early as 1 month post-transplantation. Of note, transplantation of human neurons induced a significantly more robust recovery than fetal rat striatal grafts. Thus, the logistical and ethical concerns about the use of fetal striatal cells for transplantation therapy can be eliminated by exploiting cell line-derived human neurons as alternative graft sources. Transplantation of human neurons has a therapeutic potential for treatment of behavioral deficits associated with cerebral ischemia. PMID- 9858384 TI - Material-specific lateralization of prefrontal activation during episodic encoding and retrieval. AB - Although numerous neuroimaging studies have examined the functional neuroanatomy supporting episodic memory for verbal material, there have been few investigations of non-verbal episodic encoding and retrieval. We used fMRI to directly compare prefrontal activation elicited by verbal and non-verbal material during encoding and during retrieval. Regardless of the mnemonic operation (encoding/retrieval), inferior prefrontal activation lateralized based on material type. Verbal encoding and retrieval resulted in greater left inferior prefrontal activation, whereas non-verbal encoding and retrieval resulted in greater right inferior prefrontal activation. The similarity between inferior prefrontal activity during encoding and during retrieval indicates that these mnemonic operations depend on shared processes mediated by inferior prefrontal regions. PMID- 9858382 TI - HPLC detection of serotonin within the rat cochlea. AB - This study was performed to analyse the cochlear concentrations of serotonin (5 HT) and 5-hydroxyindole-3-acetic acid (5-HIAA), their sources and modifications induced by noise exposure. Superior cervical ganglionectomy did not modify these concentrations. However, removal of the blood by aortic perfusion reduced significantly (about 76%) the cochlear concentration of 5-HT without affecting the 5-HIAA concentration. These results indicate that blood constitutes an important source of 5-HT to the cochlea, opposite to the superior cervical ganglion. Exposure to noise at 90 dB SPL did not modify the total cochlear concentrations of 5-HT and 5-HIAA, or the concentrations remaining after removal of the blood, suggesting that 5-HT could have a modulatory role in the cochlea distinct from that of olivocochlear neurotransmitters. PMID- 9858385 TI - Effects of YM872 on atrophy of substantia nigra reticulata after focal ischemia in rats. AB - Middle cerebral artery (MCA) occlusion causes atrophy in the ipsilateral substantia nigra reticulata (SNR). The effects of glutamate AMPA receptor antagonism on SNR atrophy, which is supposed to inhibit excitatory inputs from the subthalamic nucleus to the SNR, was investigated in rats with permanent MCA occlusions. Histological examination revealed marked atrophy two weeks after MCA occlusion in the saline-treated control group. However, constant i.v. infusion of YM872, a selective AMPA receptor antagonist, for 2 weeks significantly reduced SNR atrophy; neurological deficits also decreased. These results suggest that the AMPA receptor may be involved in the pathogenesis of SNR atrophy during the subacute phase of focal cerebral ischemia. PMID- 9858386 TI - Circadian modulation of membrane properties in slices of rat suprachiasmatic nucleus. AB - Perforated patch clamp recordings of neurons in slices of the suprachiasmatic nucleus (SCN) were made in their subjective day and night phases. The spontaneous firing rate and input resistance were significantly higher during the subjective day as compared to the subjective night. In parallel, the membrane potential of neurons recorded at daytime was significantly more depolarized than at nighttime. The day-night differences in membrane potential and input resistance were maintained when spikes and GABA(A) receptor-mediated transmission were blocked by tetrodotoxin and bicuculline, suggesting a cell-autonomous regulation of the circadian modulation of membrane properties. In conclusion, these findings suggest that at least one hyperpolarizing ionic conductance would be open at night and closed during mid-day, when spontaneous firing reaches peak levels in the SCN. PMID- 9858388 TI - Randomized event-related experimental designs allow for extremely rapid presentation rates using functional MRI. AB - Previous studies have shown that hemodynamic response overlap severely limits the maximum presentation rate with event-related functional MRI (fMRI) using fixed intertrial experimental designs. Here we demonstrate that the use of randomized experimental designs can largely overcome this limitation, thereby allowing for event-related fMRI experiments with extremely rapid presentation rates. In the first experiment, fMRI time courses were simulated using a fixed intertrial interval design with intervals of 16, 3, and 1 s, and using a randomized design having the same mean intertrial intervals. We found that using fixed intertrial interval designs the transient information decreased with decreasing intertrial intervals, whereas using randomized designs the transient information increased with decreasing mean intertrial intervals. In a second experiment, fMRI data were collected from two subjects using a randomized paradigm with visual hemifield stimuli presented randomly every 500 ms. Robust event-related activation maps and hemodynamic response estimates were obtained. These results demonstrate the feasibility of performing event-related fMRI experiments with rapid, randomized paradigms identical to those used in electrophysiological and behavioral studies, thereby expanding the applicability of event-related fMRI to a whole new range of cognitive neurosciences questions and paradigms. PMID- 9858387 TI - Cyclic nucleotides attenuate lipid peroxidation-mediated neuron toxicity. AB - Recent studies suggest that increased lipid peroxidation and lipid peroxidation products, such as 4-hydroxynonenal (HNE), contribute to neuronal loss in conditions associated with oxidative stress. The focus of the present study was to determine possible neuroprotective effects of elevated cyclic nucleotide levels against lipid peroxidation and HNE-mediated neural toxicity. Application of 8-bromo derivative analogs of cAMP or cGMP resulted in attenuation of HNE induced increases in mitochondrial calcium, reactive oxygen species, and neuron loss. Similar results were obtained when neural cells were pretreated with the phosphodiesterase inhibitors zaprinast or isobutylmethylxanthanine (IBMX). These data are consistent with a possible neuroprotective role for elevated cyclic nucleotide levels in disorders associated with increases in lipid peroxidation and HNE. PMID- 9858389 TI - Time course of the medial olivocochlear efferent effect on otoacoustic emissions in humans. AB - The time course of the medial olivocochlear efferent system has been studied in humans, using the suppressive effect of a contralateral broad-band noise (CBBN) on 2f1-f2 distortion product otoacoustic emissions (DPOAE) amplitude. DPOAE were recorded with F2 at 1.5 kHz, with a temporal resolution of 2.6 s, in the presence and absence of a 40 dB SL continuous CBBN, whose duration ranged between 30 s and 20 min. The CBBN suppressive effect on DPOAE amplitude was greatest from the first 2.6 s, and was sustained for 20 min. At the CBBN offset, when the CBBN duration was > or = 2 min, DPOAE amplitude continued to increase for > 1 min, showing an efferent effect outlasting CBBN stimulation. PMID- 9858390 TI - Identification of olfactory receptor mRNA sequences from the rat olfactory bulb glomerular layer. AB - In situ hybridization has demonstrated mRNA for olfactory receptors (OR) in the axon terminals of olfactory receptor neurons. Neurons that express the same OR appear to send their axons to two stereotyped glomeruli in the olfactory bulb (OB). Based on these observations, we tested the feasibility of using RT-PCR to isolate and sequence OR mRNA from small samples of the rat OB glomerular layer. Biomagnetic mRNA isolation followed by RT-PCR yielded partial sequences for 21 novel members of the OR family. The results suggest that the topography of OR mRNA can be mapped across the OB, to study synaptic specificity and odor representation in the olfactory system. PMID- 9858391 TI - CNTF and GDNF, but not NT-4, support corticospinal motor neuron growth via direct mechanisms. AB - Axotomy and neurodegenerative diseases cause corticospinal motor neuron (CSMN) degeneration. We previously showed that CNTF, NT-4 and GDNF can support CSMN survival in enriched preparations. Here we developed a fluorescence-activated cell sorting method to highly purify CSMN (97+/-4.6%). We tested the neurotrophic activities of CNTF, NT-4 and GDNF on enriched and purified CSMN preparations. Similar to their effects on enriched CSMN preparations, CNTF and GDNF sustained the survival of purified CSMN for at least 5 days with ED50 values of 1.28+/-0.46 nM and 0.59+/-0.39 nM, respectively. In contrast, NT-4 supported survival of enriched but not of purified CSMN, indicating that CNTF and GDNF sustain motor neuron survival by direct action of CSMN, while NT-4 requires accessory cells present in enriched CSMN preparations. PMID- 9858392 TI - Monoclonal antibodies reveal expression of the CGRP receptor in Purkinje cells, interneurons and astrocytes of rat cerebellar cortex. AB - The molecular layer of adult rat cerebellum displays high levels of calcitonin gene-related peptide (CGRP) receptors, but the cellular location of the receptor remains unidentified. In an attempt to reveal the expression sites of these receptors, monoclonal antibodies raised against purified CGRP receptors from porcine cerebellar membranes were used in double-immunofluorescence experiments combined with confocal microscopy. PEP-19, a marker that is highly enriched in Purkinje cells (Pc), revealed that CGRP receptors are located in Pc cytoplasm and dendrites, where they label small puncta sometimes arranged in a row along the course of the dendrite itself. CGRP receptors were also located in inhibitory interneurons. Furthermore, as shown by double-labeling experiments with GFAP, CGRP receptor-IR labeled Golgi epithelial cells and their radial fibers (Bergmann fibers), as well as astrocytic processes encircling Pc somata. The simultaneous presence of CGRP receptors in Purkinje cells and in the glial cells that heavily enshroud Purkinje cells allows us to hypothesize that these receptors may be involved in neuron-glia interactions influencing neuronal activity. PMID- 9858393 TI - Effects of exercise and training on blood rheology. AB - The effects of exercise on the rheological properties of blood have not received much research attention. Recent, limited evidence indicates that the viscosities of whole blood and plasma increase in response to a variety of exercise protocols. The increase in whole blood viscosity is mainly attributed to an increase in haematocrit and plasma viscosity, whereas the deformability and aggregability of red blood cells remain unaltered. The increases in plasma viscosity and haematocrit have been ascribed to exercise-induced haemoconcentration as a result of fluid transfer from the blood to the interstitial spaces. Although the long term effects of endurance training on blood rheology have been very briefly examined, the exact effect of training has not as yet been determined. However, available cross-sectional and longitudinal studies indicate that the blood of endurance athletes is more dilute and this has been attributed to an expansion of plasma volume as a result of training. It has been suggested that this blood dilutional effect of endurance training may be advantageous in delivering oxygen to the exercising muscles because of a reduced resistance to blood flow. The increase in plasma volume may also contribute to the body water pool and help offset dehydration. The influence of strength and power training on blood rheology is not known. PMID- 9858395 TI - Applied physiology of water polo. AB - Water polo has been played for over a century. While the rules of the game have evolved considerably over this time, the sport has consistently remained, physiologically, a highly demanding activity. Much attention has been paid to the technical and strategic elements of the game; however, despite the potential for improvements in athletic performance and the maintenance of athletes' health, there are few published studies (particularly in English) on the physical and physiological demands and adaptations to water polo training and competition. Game analyses have demonstrated that water polo is an 'intermittent' sport comprised of intense bursts of activity of <15 seconds duration with intervening, lower intensity intervals averaging <20 seconds duration. Physiological measurements obtained during game play indicate a cumulative effect of the repeated sequences of activities and suggest there is a high metabolic demand on the athletes. The multiple individual skills and movements required for playing water polo also place considerable demands on the neuromuscular system. Observations of the frequency and duration of the different activities, and of the physiological responses to participating in a water polo match, are initial sources of information for designing training programmes specific to the game and to the different playing positions. The physical and physiological attributes of elite water polo players offer some insight into the minimum requirements for participation and the adaptations that result from training and competition. Further systematic documentation and experimentation are required to facilitate the design and specification of individual training programmes and to better understand the long term effects of water polo on athletes' health. PMID- 9858396 TI - Sudden death in athletes: an update. AB - The athlete projects the ultimate image of well-being in the health status spectrum. Nevertheless, exercise-related sudden cardiac death (SCD) is an uncommon, yet tragic, occurrence. Exercise-related SCD is defined by symptoms that arise within 1 hour of participation in sport. The major mechanisms involved in exercise-related SCD are related to haemodynamic and electrophysiological changes brought about by exercise in the susceptible individual. Fatal arrhythmia seems to be the most common mechanism of death. Between 1 and 5 cases of SCD per 1 million athletes occur annually. In young athletes (<35 years old), the majority of these cases are caused by defined and hereditary cardiovascular disorders. Among other aetiologies, hypertrophic cardiomyopathy and coronary artery anomalies are most common in this group. In older athletes (>35 years old), sudden death is usually associated with atherosclerotic cardiac disease. A problem for identifying athletes at risk for SCD is that the athlete's heart undergoes adaptive changes in response to regular physical exercise. Alterations in cardiac function influence the physical examination, the electrocardiogram and the echocardiogram. Because of these characteristic 'abnormalities' of the athlete's heart, it is often difficult to distinguish physiological adaptations from pathophysiological processes. Although studies and observations have helped to clarify the cardiovascular pathology responsible for SCD in young, apparently healthy individuals, effective methods for preventing SCD and identifying and screening athletes at risk remain elusive. Problems with routine comprehensive screening of athletes include the limitations inherent in the predictive value of available diagnostic procedures and the cost of testing large populations. The variation from normal cardiac physiology found within the athletic population and the rarity of SCD in athletes means that elaborate screening to determine individuals at risk is neither practical nor cost effective. A thorough assessment of pertinent family and medical histories, cardiac auscultation of young athletes, evaluation of exercise-induced symptoms and education of older athletes to the symptoms of cardiac ischaemia are all essential to primary prevention of SCD in the athletic population. Until reliable methods can accurately identify those athletes at risk for SCD, broad recommendations are available to help guide the management and participation in sports of athletes with cardiovascular disease. PMID- 9858397 TI - Axillary nerve injuries in contact sports: recommendations for treatment and rehabilitation. AB - Axillary nerve injuries are some of the most common peripheral nerve injuries in athletes who participate in contact sports. Resulting deltoid muscle paralysis is secondary to nerve trauma which occurs following shoulder dislocation or a direct blow to the deltoid muscle. Compression neuropathy has been reported to occur in quadrilateral space syndrome as the axillary nerve exits this anatomic compartment. The axillary nerve is also extremely vulnerable during any operative procedure involving the inferior aspect of the shoulder, and iatrogenic injury to the axillary nerve remains a serious complication of shoulder surgery. Accurate diagnosis of axillary nerve injury is based on a careful history and physical examination as well as an understanding of the anatomy of the shoulder and the axillary nerve in particular. Inspection, palpation and neurological testing provide the bases for diagnosis. A clinically suspected axillary nerve injury should be confirmed by electrophysiological testing, including electromyography and nerve conduction studies. During the acute phase of injury, the athlete should be rested and any ligamentous or bony injury should be treated as indicated. Patients should undergo an extensive rehabilitation programme emphasising active and passive range of motion as well as strengthening of the rotator cuff, deltoid and periscapular musculature. Shoulder joint contracture should be avoided at all costs as a loss of shoulder mobility may ultimately affect functional outcome despite a return of axillary nerve function. If no axillary nerve recovery is observed by 3 to 4 months following injury, surgical exploration is indicated. Athletes who sustain injury to the axillary nerve have a variable prognosis for nerve recovery, although the return of function of the involved shoulder is typically good to excellent. We recommend that athletes who sustain axillary nerve injury may return to contact sport participation when they achieve full active range of motion of the shoulder and when shoulder strength is documented to be good to excellent by isometric or manual muscle testing. PMID- 9858398 TI - Evaluation of automated methods of measuring hemoglobin and hematocrit in horses. AB - OBJECTIVE: To evaluate the accuracy of 3 automated methods of determining Hct and hemoglobin (Hb) concentration, compared with manual methods. Animals-22 clinically normal adult horses of various breeds. PROCEDURE: A blood sample was obtained from each horse. Six dilutions (representing Hct of 0, 10, 20, 40, 60, or 70%) were prepared from each sample and analyzed, using 1 of 2 blood gas analyzers or a hemoximeter (for automated determinations) or the Wintrobe macrohematocrit and cyanmethemoglobin methods (for manual determinations). Regression analysis was used to determine mean slope relationships between Hct and Hb measurements obtained by use of manual versus automated methods. Slopes were compared, using Student's t-test. RESULTS: Of the 3 automated methods examined, only 1 blood gas analyzer reported Hct and Hb values that were not significantly different from those determined by use of manual methods; however, this analyzer could not report Hb concentrations below 2.5 g/dl. The other blood gas analyzer reported values for Hct and Hb concentrations that were consistently higher than those obtained by use of manual methods at Hct < or = 20% and Hb < or = 6.6 g/dl. The hemoximeter yielded more accurate results if the Hb concentration was between 6.6 and 20 g/dl. CONCLUSION: Although there were some limitations in measuring at low Hb concentrations, the method of determining Hb concentration and Hct with blood gas analyzer 2 was more accurate than that with blood gas analyzer 1 (Hct and Hb concentration) or the hemoximeter (Hb only). PMID- 9858399 TI - Reproducibility of breath hydrogen concentration measurements in dogs after change of diet. AB - OBJECTIVE: To assess reproducibility of breath hydrogen (H2) concentration measurements in dogs on 3 consecutive days after introduction of a new diet. ANIMALS: 10 healthy colony-source dogs. Procedure Dogs were fed a commercial pet food for a minimum of 10 days before introduction of a high-carbohydrate commercial diet. Expired breath samples were collected at baseline and every 90 minutes for 18 hours on the first 3 days after introduction of the new diet. The area under the curve (AUC) for breath H2 concentration versus time on days 1, 2, and 3 were compared by use of repeated-measures ANOVA. RESULTS: The (AUC) did not differ significantly among test days 1, 2, and 3. CONCLUSIONS: Expired breath H2 concentration in dogs indicated good reproducibility during the first 3 days after introduction of a new diet. CLINICAL RELEVANCE: It is not necessary to provide a period for adaptation to a test diet prior to performing breath H2 tests in dogs. PMID- 9858400 TI - Evaluation of an assay for detecting telomerase activity in neoplastic tissues of dogs. AB - OBJECTIVE: To determine feasibility of using the telomere repeat amplification protocol (TRAP) assay to detect telomerase activity in tumors of dogs. SAMPLE POPULATION: Samples of tumor or normal tissue were obtained from client-owned dogs that underwent surgical biopsy during the period of January 1996 through December 1997. PROCEDURE: The TRAP assay was used to detect telomerase activity in malignant or benign tumors of dogs. Telomerase status (positive or negative) was compared with results of histologic examination for each sample to estimate specificity and sensitivity of this assay for the diagnosis of malignancy. RESULTS: Of 26 malignant tumors, 24 were telomerase positive on TRAP assay, whereas 3 of 4 benign tumors and 3 of 3 normal tissues were telomerase negative. Analysis of these results indicated an estimated sensitivity of 92% and specificity of 86% for tumor analysis, using the TRAP assay. CONCLUSION: The TRAP assay can be used to measure telomerase activity in malignant tumors of dogs. CLINICAL RELEVANCE: Because telomerase activation may be required for indefinite longevity of cells, it may also serve as a tumor marker and therapeutic target. The TRAP assay can be used to detect telomerase in samples of fluid as well as tissues obtained from solid tumors. Therefore, it may have considerable clinical value in rapid and noninvasive diagnosis of neoplasia in dogs. Additional studies must be completed to more accurately determine sensitivity and specificity of the assay. PMID- 9858394 TI - Associations between physical activity and susceptibility to cancer: possible mechanisms. AB - Physical activity is associated with a reduced risk of all-cause and colonic cancers, and it seems to exert a weaker effect on the risk of breast, lung and reproductive tract tumours. This review examines possible mechanisms behind the observed associations. Restriction of physical activity by pre-existing disease may contribute to the association with lung cancers, but seems a less likely explanation for other types of tumour. Indirect associations through activity related differences in body build or susceptibility to trauma seem of minor importance. Potential dietary influences include overall energy balance and energy expenditure, the intake and/or bioavailability of minerals, antioxidant vitamins and fibre, and the relative proportions of protein and fat ingested. Links between regular exercise and other facets of lifestyle that influence cancer risks are not very strong, although endurance athletes are not usually smokers, and regular leisure activity is associated with a high socioeconomic status which tends to reduce exposure to airborne carcinogens, both at work and at home. Overall susceptibility to cancer shows a 'U'-shaped relationship to body mass index (mass/height2) reflecting, in part, the adverse influences of cigarette smoking and a tall body build for those with low body mass indices and, in part, the adverse effect of obesity at the opposite end of the body mass index distribution. Obesity seems a major component in the exercise-cancer relationship, with a particular influence on reproductive tract tumours; it alters the pathways of estradiol metabolism, decreases estradiol binding and facilitates the synthesis of estrogens. Among the hormonal influences on cancer risk, insulin-like growth factors promote tumour development and exercise mediated increases in cortisol and prostaglandin levels may depress cellular components of immune function. However, the most important change is probably the suppression of the gonadotropic axis. Apparent gender differences in the benefits associated with regular exercise reflect gender differences in the hormonal milieu and also a failure to adapt activity questionnaires to traditional patterns of physical activity in females. The immune system is active at various stages of tumour initiation, growth and metastasis. However, acute and chronic changes in immune response induced by moderate exercise are rather small, and their practical importance remains debatable. At present, the oncologist is confronted by a plethora of interesting hypotheses, and further research is needed to decide which are of practical importance. PMID- 9858401 TI - Relations between biochemical markers of osteoarthritis and other disease parameters in a population of dogs with naturally acquired osteoarthritis of the genual joint. AB - OBJECTIVE: To investigate the hypothesis that concentrations of the keratan sulfate epitope, 1/20/5D4 (5D4) and total sulfated glycosaminoglycans (S-GAG) in synovial fluids, and of 5D4 in serum of dogs with naturally acquired osteoarthritis of the genual joint (stifle), secondary to cranial cruciate ligament deficiency, are associated with other disease parameters of osteoarthritis. ANIMALS: 58 dogs with stifle osteoarthritis secondary to naturally acquired cranial cruciate ligament (CCL) deficiency. PROCEDURE: All dogs were examined clinically, radiographically and, in some instances, scintigraphically. Serum and synovial fluid from both stifles were assayed for 5D4 concentration, using an inhibition ELISA, and for S-GAG, using a direct dye binding assay. RESULTS: Serum 5D4 concentration was not significantly associated with other disease features. Total S-GAG values were high, and 5D4 values were low, in synovial fluid from clinically active, compared with contralateral joints. The S-GAG concentration in synovial fluid from clinically active joints was negatively correlated with radiographic severity score (rs = -0.389, P = 0.004, Spearman's rank correlation). Other associations between marker concentrations and disease parameters could not be detected. CONCLUSIONS: Serum 5D4 concentration is not a useful marker of stifle osteoarthritis in dogs; however, synovial fluid S-GAG and 5D4 values may hold more promise as disease measures. PMID- 9858402 TI - Serotyping scheme for Staphylococcus aureus isolated from cows with mastitis. AB - OBJECTIVE: To identify the Staphylococcus aureus capsular serotypes that are not typable, using capsular serotypes 5 and 8, which are currently used to type S aureus isolated from cows with mastitis. SAMPLE POPULATION: Milk samples (n = 273) from cows with mastitis in 178 dairy herds in California, Wisconsin, Michigan, Texas, and New York that were collected by state diagnostic laboratories and S aureus-positive milk samples collected by Veterinary Health Services in the United Kingdom (15), France (22), The Netherlands (36), and Germany (21). PROCEDURE: Capsular serotyping of coded isolates was performed by use of direct cell agglutination and immunoprecipitation of cell extracts with antisera specific for capsular types 5 and 8 and a newly developed S aureus serotyping antiserum 336. RESULTS: In the United States, S aureus capsular types 5 and 8 accounted for 18 and 23% of the isolates, respectively, and type 336 accounted for 59%. Percentage of capsular serotypes in European samples were as follows: type 5 = 34%, type 8 = 34%, type 336 = 30%, and nontypable = 2%. CONCLUSIONS: Serotypes 5 and 8 accounted for only 41% of S aureus isolates from US milk samples, but accounted for 70% of isolates from European milk samples. Addition of the newly developed serotyping antiserum 336 to the typing scheme accounted for 100% of US samples and 98% of European samples and will enable development of a more comprehensive S aureus vaccine. PMID- 9858403 TI - Clinical consequences of exposing pregnant gilts to strains of porcine reproductive and respiratory syndrome (PRRS) virus isolated from field cases of "atypical" PRRS. AB - OBJECTIVE: To determine clinical consequences of exposing pregnant gilts to strains of porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) isolated from field cases of "atypical" or "acute" PRRS in vaccinated herds. ANIMALS: 20 pregnant gilts and their pigs and fetuses. PROCEDURE: 8 pregnant gilts (principals: 4 groups [2 gilts/group]) were exposed oronasally at or about 45 days of gestation to 1 of 4 strains of PRRSV and necropsied 6 weeks later. Nonexposed controls (2 additional pregnant gilts) were kept under otherwise similar conditions. The experiment was repeated, except that principals were exposed at or approximately 90 days of gestation and allowed to farrow. Clinical observations were made at least twice daily, and samples and specimens from gilts and their fetuses and pigs were tested for PRRSV and homologous antibody. RESULTS: Exposure of pregnant gilts to PRRSV at or approximately 45 days' gestation resulted in low prevalence of transplacental infection and fetal death. Exposure of pregnant gilts to PRRSV at or approximately 90 days' gestation resulted in higher prevalence of transplacental infection and fetal death. Moreover, 1 gilt aborted and many liveborn pigs of other litters were weak and unthrifty. Clinical signs of disease and reproductive failure were especially severe for a field strain of PRRSV isolated from an epizootic that fit the strictest definition of atypical PRRS. Controls remained clinically normal and free of PRRSV. CONCLUSION AND CLINICAL RELEVANCE: Some strains of PRRSV now circulating in US swine herds are more virulent than those encountered in the past. Clinical PRRS in vaccinated herds suggests need for a new generation of vaccines. PMID- 9858404 TI - Hoof size, shape, and balance as possible risk factors for catastrophic musculoskeletal injury of Thoroughbred racehorses. AB - OBJECTIVE: To evaluate hoof size, shape, and balance as risk factors for catastrophic musculoskeletal injuries (CMI), including suspensory apparatus failure (SAF) and cannon bone condylar fracture (CDY) in Thoroughbred racehorses. ANIMALS: 95 Thoroughbred racehorses that died between 1994 and 1996. PROCEDURE: 38 quantitative measures of hoof size, shape, and balance were obtained from orthogonal digital images of the hoof and were compared between case horses with forelimb CMI (70), SAF (43), and CDY (10) injuries and control horses whose death was unrelated to the musculoskeletal system (non-CMI, 25). Comparison of group means between cases and controls was done using ANOVA, and multivariable logistic regression was used to estimate odds ratios. RESULTS: Odds of CMI were 0.62 times lower for a 5mm increase in ground surface width difference and 0.49 times lower for a 100-mm2 increase in sole area difference. Odds of SAF were 6.75 times greater with a 10 degrees increase in toe-heel angle difference and 0.58 times lower with a 100-mm2 increase in sole area difference. Odds of CDY were 0.26 times lower with a 3 degrees increase in toe angle, 0.15 times lower with a 5-mm increase in lateral ground surface width, and 0.35 times lower with a 100-mm2 increase in sole area difference. CLINICAL RELEVANCE: Decreasing the difference between toe and heel angles should decrease risk of SAF for Thoroughbred racehorses and should be considered in addition to increasing toe angle alone to help prevent catastrophic injury. Trimming the hoof to perfect mediolateral symmetry may not be a sound approach to avoiding injury. PMID- 9858405 TI - Prediction of inherited portosystemic shunts in Irish Wolfhounds on the basis of pedigree analysis. AB - OBJECTIVE: To test validity of prediction for inherited portosystemic shunts (PSS) in Irish Wolfhounds, using nonselective clinical findings and a computerized database containing 5-generation pedigrees. ANIMALS: 613 dogs in the first and 396 dogs in the second cohort. PROCEDURE: Preprandial venous ammonia concentration was measured at 6 to 8 weeks in all pups born between Jan 1, 1988 and Jan 1, 1997 Portosystemic shunts were confirmed in hyperammonemic pups, using radioisotope shunt index measurement, and diagnosis of shunting was confirmed at abdominal surgery or necropsy. Findings in dogs of the first cohort (born before Jan 1, 1992) were used to predict shunting in their offspring of the second cohort. Common ancestors of first-cohort dogs with shunts were tested for positive associations with the disease. Risk for a shunt in all second-cohort dogs was predicted on the basis of relatedness with founders and was compared with outcome of clinical screening. RESULTS: Prevalence of shunts in first and second cohorts was 3.1 and 2.3%, respectively. Fifteen highly related associated founders could be identified. Second-cohort dogs were classified into 6 groups of increasing predicted risk. Mean number of dogs per class was 60; number of clinically diagnosed cases ranged from 0 in the class with the lowest risk to 4 in the highest risk class. CONCLUSIONS: Genetic risk for reproducing a PSS in Irish Wolfhounds was accurate, using the described method. CLINICAL RELEVANCE: Risk estimation provides a tool for genetic counseling, does not require knowledge of the mode of inheritance, and may be valid for any inherited disease. PMID- 9858406 TI - Molecular characteristics of equine stromelysin and the tissue inhibitor of metalloproteinase 1. AB - OBJECTIVE: To clone the entire coding sequence of equine matrix metalloproteinase 3 (MMP-3, stromelysin) and tissue inhibitor of metalloproteinase-1 (TIMP-1) and compare their nucleotide and amino acid sequences with those of MMP-3 and TIMP-1 from other species. SAMPLES: Articular cartilage harvested from the joints of 4 foals, 2 yearlings, and 3 adult horses. PROCEDURE: A cDNA library was constructed from mRNA extracted from equine chondrocytes. The library was screened and clones selected that contained the cDNA for MMP-3 and TIMP-1. The cDNA was sequenced and the nucleotide and deduced amino acid sequences compared with known sequences in other species. Northern blot analysis was performed, using the resulting cDNA clones. RESULTS: An 1803-bp cDNA for MMP-3 including the entire coding sequence of 1434 bases was cloned and sequenced. A 744-bp cDNA for TIMP-1 including the entire coding sequence of 624 bases was cloned and sequenced. Northern analysis revealed MMP-3 to hybridize to a single mRNA species at approximately 2.1 kb. TIMP-1 hybridized to a single mRNA species at approximately 0.8 kb. CONCLUSIONS: MMP-3 and TIMP-1 were highly homologous to that of other species at the nucleotide and amino acid level although each had unique residues in part of the peptide that is generally conserved. CLINICAL RELEVANCE: Understanding the molecular structure of MMP-3 and TIMP-1 and the availability of their cDNA should allow a more detailed understanding of their balance in cartilage and the degradative processes in joint disease. PMID- 9858407 TI - Interleukin 8 response by bovine mammary epithelial cells to lipopolysaccharide stimulation. AB - OBJECTIVE: To determine whether an established bovine mammary epithelial cell line expresses interleukin 8 (IL-8) mRNA and synthesizes antigenic IL-8 in response to lipopolysaccharide (LPS) stimulation. SAMPLE POPULATION: A bovine mammary epithelial cell line (MAC-T). PROCEDURE: mRNA was isolated from cells stimulated with graded concentrations of LPS. The first strand of IL-8 cDNA was synthesized, using a reverse transcriptase (RT) reaction with a specific oligonucleotide. Amplification of IL-8 cDNA was obtained by use of polymerase chain reaction (PCR). The MAC-T-derived antigenic IL-8 was quantified by use of a commercial anti-human IL-8 kit in a sandwich ELISA. RESULTS: RT-PCR revealed expression of MAC-T-derived mRNA within the first hour after stimulation with LPS. Expression of IL-8 mRNA was correlated to production of IL-8 protein detected in medium by use of the sandwich ELISA. Amounts of antigenic IL-8 increased in a dose- and time-dependent manner, and were maximal (57 pg/ml) at 48 hours after stimulation with 20 microg of LPS/ml. CONCLUSIONS: MAC-T cells secrete IL-8 in response to stimulation with LPS in a dose- and time- dependent manner. The results were consistent with our hypothesis that mammary gland epithelial cells can be a source of IL-8 during the early stage of mastitis. Therefore, IL-8 may have a pivotal role in resolving bacterial infections. PMID- 9858408 TI - Effects of human intravenous immunoglobulin on canine monocytes and lymphocytes. AB - OBJECTIVE: To evaluate interactions of human intravenous immunoglobulin (IVIG) with canine lymphocytes and monocytes. SAMPLE POPULATION: Heparinized blood samples from 4 clinically normal Beagles. PROCEDURE: Binding ability of IVIG to canine lymphocytes and monocytes was measured by flow cytometry and an indirect immunofluorescent assay. Dual-staining fluorescent assays were done to determine lymphocyte subsets that bind IVIG. Competitive assays were done, using intact canine IgG and Fc fragments, and inhibition of binding was compared with that of F(ab)2 fragments. Ability of IVIG to inhibit phagocytosis of antibody-coated canine RBC also was determined, using a canine mononuclear cell phagocytic assay. RESULTS: IVIG concentrations (10, 1, 0.1, and 0.01 mg/ml) bound to (mean+/-SD) 99.6+/-0.4, 92.4+/-6.1, 20.4+/-24.6 and 2.0+/-5.1 % of canine lymphocytes, respectively, Dual staining analyses with IVIG and canine lymphocyte markers indicated that IVIG bound to CD4, CD8, and B lymphocytes. The aforementioned 4 IVIG concentrations bound to 98.0+/-2.1, 85.5+/-13.5, 64.7+/-32.8, and 26.5+/ 17.1 % of monocytes, respectively. Inhibition of IVIG (0.01 mg/ml) binding to monocytes was significant (P< 0.05) in the presence of 1 and 10 mg of canine IgG/ml and 1 mg of canine Fc fragments/ml. In the presence of F(ab')2 fragments of canine IgG, inhibition was not significant, suggesting that binding is Fc mediated. Co-culturing of monocytes, opsonized RBC, and the 4 concentrations of IVIG and no IVIG resulted in 11.8+/-5.1, 27.7+/-12.3, 31.8+15.1, 53.8+/-6.7, and 45 + 12% of the monocytes containing RBC, respectively. Phagocytosis inhibition was significant (P < 0.05) at an IVIG concentration of 10 mg/ml. CONCLUSIONS: IVIG binds to canine lymphocytes and monocytes; binding to the latter is Fc mediated. IVIG also inhibits Fc-mediated phagocytosis of antibody-coated RBC. CLINICAL RELEVANCE: Owing to its ability to inhibit Fc-mediated phagocytosis of antibody-coated RBC, IVIG may be an effective short-term treatment for dogs with immune-mediated hemolytic anemia. PMID- 9858409 TI - Protection against infection and abortion induced by virulent challenge exposure after oral vaccination of cattle with Brucella abortus strain RB51. AB - OBJECTIVES: To determine efficacy of orally administered Brucella abortus vaccine strain RB51 against virulent B abortus challenge exposure in cattle as a model for vaccination of wild ungulates. ANIMALS: 20 mixed-breed beef cattle obtained from a brucellosis-free herd. PROCEDURE: Sexually mature, Brucella-negative beef heifers were vaccinated by mixing > 10' viable RB51 organisms or diluent with their feed. Heifers were fed individually and consumed their entire ration. Each heifer received approximately 3 X 10' colony-forming units (CFU). Six weeks after oral vaccination, heifers were pasture-bred to brucellosis-free bulls. At approximately 186 days' gestation, heifers were challenge exposed conjunctively with 107 CFU of virulent B abortus strain 2308. RESULTS: Vaccination with the rough variant of B abortus RB51 did not stimulate antibodies against the O polysaccharide (OPS) of B abortus. After challenge exposure and parturition, strain 2308 was recovered from 80% of controls and only 20% of vaccinates. Only 30% of the vaccinates delivered dead, premature, or weak calves, whereas 70% of the controls had dead or weak calves. CONCLUSIONS: Cattle vaccinated orally with the rough variant of B abortus strain RB51 develop significant (P < 0.05) protection against abortion and colonization and do not produce OPS-specific antibodies. Clinical Relevance-Results encourage further investigation into use of strain RB51 to vaccinate wild ungulates (elk and bison) orally. PMID- 9858410 TI - Survey for antibodies to bovine adenoviruses in six- to nine-month-old feedyard cattle. AB - OBJECTIVE: To determine the prevalence of antibody to bovine adenovirus (BAdV) serotypes 1-8 and 10 in calves at a farm and after 5 weeks in a feedyard. ANIMALS: 2- to 5-month-old calves of mixed English breeding (n = 100) from 4 farms. PROCEDURE: Serum BAdV antibody was measured by use of a microtitration test. RESULTS: Serum antibodies were found to the 9 BAdV serotypes studied. Seroconversion to each virus had occurred in some calves by the time the second serum sample had been obtained, indicating that the BAdV were present and inducing active infection in these calves. CONCLUSIONS: Antibody to BAdV serotypes 1-8 and 10 are present in cattle populations of the United States, indicating existence of these serotypes, although only BAdV serotypes 1-4, 7, and 10 have been isolated. PMID- 9858411 TI - Molecular, clinical, and pathologic comparison of two distinct strains of Haemobartonella felis in domestic cats. AB - OBJECTIVE: To characterize 2 strains of Haemobartonella felis by use of molecular techniques. ANIMALS: 35 specific-pathogen-free cats, 6 months to 4 years old. PROCEDURE: Intraperitoneal or IV inoculation with blood containing H felis small form (Hfsm, 18 cats) or H felis large form (Hflg, 11 cats); 6 cats were uninfected controls. Hfsm was evaluated for capability to cross-protect against the more virulent Hflg. Morphology of both strains was compared by light microscopy of Wright-Giemsa-stained blood smears, and the 16S rRNA genes were sequenced. RESULTS: Infection with Hflg induced signs of depression, fever, and severe macrocytic normochromic anemia with nucleated erythrocytes. More than 95% of erythrocytes were parasitized. Inoculation with Hfsm and uninfected control blood induced mild or no clinical signs and no hematologic abnormalities. Anti-H felis IgG was first detected on postinoculation day (PID) 21, and increased to maximal titer of 400 by PID 28. Reactivated infection was observed in 8 of 29 cats (4 Hfsm and 4 Hflg), with 5% parasitized erythrocytes during the later attack. On PID 8, Hflg-inoculated cats had positive results of polymerase chain reaction analysis (PCR) that persisted until cats were treated with doxycycline or oxytetracycline; Hfsm-inoculated cats had positive PCR results that persisted for duration of observation (3 months). CONCLUSIONS: Genetically and morphologically distinct strains of H felis infect cats in the field. The level of genetic difference suggested that these strains may be different species or genera. CLINICAL RELEVANCE: PCR is a critical diagnostic aid to detect occult Haemobartonella spp infection, as well as response to treatment and clearance of the organism. PMID- 9858412 TI - Population pharmacokinetics of gentamicin in horses. AB - OBJECTIVE: To develop and validate a population pharmacokinetic model for gentamicin in horses, using retrospective clinical data. ANIMALS: 62 horses that had been treated IV with multiple doses of gentamicin at our veterinary teaching hospital between 1987 and 1996. Procedure-46 horses were assigned to the study group, and 16 to the validation group. Detailed history of dosage, sample collection times, and selected pathophysiologic variables were recorded for each patient. Samples were analyzed by use of a fluorescence polarization immunoassay method. Pharmacostatistical analysis was conducted, using computer software. The predictive model correlates pharmacokinetic parameters to concomitant pathophysiologic variables and estimates the inter- and intraindividual variability in disposition. RESULTS: A two-compartment model best described the data. Clearance (CI) was linearly correlated to body weight and serum creatinine concentration. Volume of the central compartment (Vd(c)) was linearly related to body weight. Interindividual coefficients of variability for CI and Vd(c) were 24 and 16%, respectively. The residual variability (intraindividual) was 13%; mean prediction error percent (bias) was 2%; and mean absolute prediction error percent (precision) was 29%. CONCLUSIONS: Population pharmacokinetic analysis allows study of the basic features of gentamicin disposition in horses with sparse data per individual. A considerable proportion of the pharmacokinetic variability of gentamicin in our study population was explained by differences in body weight and serum creatinine concentration. CLINICAL RELEVANCE: Population pharmacokinetics can be used to design first-dosage regimens according to the clinical characteristics of individual animals. Population pharmacokinetic models could also be included in Bayesian forecasting strategies to improve plasma concentration predictions in individual patients. PMID- 9858413 TI - Determination of skin concentrations of enrofloxacin in dogs with pyoderma. AB - OBJECTIVE: To compare serum and skin concentrations of enrofloxacin in dogs with pyoderma with those of clinically normal dogs and to evaluate concentrations in dogs with superficial versus deep pyoderma. ANIMALS: 16 clinically normal dogs and 16 dogs with pyoderma. PROCEDURE: Enrofloxacin (approx 5 mg/kg of body weight, PO) was administered daily to all dogs. Serum samples and skin biopsy specimens were obtained on day 1 at 3 hours after drug administration and on day 3 immediately before and 3 hours after drug administration. Samples and specimens were assayed by high-performance liquid chromatography. Morphometric analysis was performed on skin biopsy specimens to determine correlation between inflammatory cells and peak tissue enrofloxacin concentration on day 1. RESULTS: Morphometric analysis revealed high correlation between dermal inflammatory cell count and drug concentration in dogs with pyoderma. CONCLUSIONS: At mean dosage of 5 mg/kg once daily, enrofloxacin tissue concentrations were significantly greater in dogs with pyoderma at 3 hours after pill administration. Enrofloxacin tissue concentration on day 3 at 3 hours after pill administration was 12.4 times the 90% minimum inhibitory concentration of enrofloxacin for Staphylococcus intermedius. CLINICAL RELEVANCE: In dogs with pyoderma, therapeutic tissue concentrations of enrofloxacin are reached as early as 3 hours after drug administration. PMID- 9858414 TI - Effects of exercise and EDTA administration on blood ionized calcium and parathyroid hormone in horses. AB - OBJECTIVE: To determine effects of exercise on blood ionized calcium (Ca2+) and plasma parathyroid hormone (PTH) concentrations in horses and to compare the effects of exercise-induced and EDTA-induced hypocalcemia on PTH secretion. ANIMALS: 17 horses entered in a show jumping competition and 5 horses given EDTA. PROCEDURE: Blood Ca2+ and plasma PTH concentrations were measured before and after exercise in the 17 horses entered in the jumping competition. In the other 5 horses, concentrations were measured during infusion of EDTA IV. RESULTS: Exercise resulted in a significant decrease in blood Ca2+ concentration and a significant increase in plasma PTH concentration, and blood Ca2+ concentration was correlated with plasma PTH concentration. Administration of EDTA resulted in hypocalcemia and an increase in PTH concentration. For the same decrease in Ca2+ concentration, magnitude of the exercise-induced increase in PTH concentration was similar to magnitude of the EDTA-induced increase. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the increase in plasma PTH concentration in horses after exercise is secondary to hypocalcemia and that the increase in PTH concentration seems to be commensurate with the decrease in Ca2+ concentration. PMID- 9858415 TI - Correlation between longitudinal bone growth, growth hormone, and insulin-like growth factor-I in prepubertal dogs. AB - OBJECTIVE: To determine the association between longitudinal bone growth and concentrations of growth hormone (GH) and insulin-like growth factor-I (IGF-I) in serum from prepubertal dogs. Animals-6 male 14-week-old German Shepherd Dogs. PROCEDURE: Blood was obtained every 30 minutes for 14 consecutive days. Concentrations of GH and IGF-I in serum were determined, using a canine-specific radioimmunoassay and conventional radioimmunoassay after acid-ethanol extraction, respectively. Simultaneous biplanar radiography was performed daily to measure bone growth. Spectral analysis was used to estimate specific features of GH secretion during an extended period. Multiple linear regression with different lag times between independent and dependent variables was used to determine the strongest predictors of bone growth. RESULTS: The power spectra of GH concentrations in serum had a primary peak at a frequency of 0.02 cycles/h or a periodicity of 50 h/cycle. A significant determinant of longitudinal bone growth was a lag time of 1 day in concentration of GH in serum. The relationship between IGF-I concentration in serum and bone growth was not significant. CONCLUSIONS: The primary frequency of GH secretion is outside the time frame of a single day and the concentration of GH in serum is a primary determinant of bone growth. CLINICAL RELEVANCE: A better understanding of the components of bone growth provide discernment to improved diagnosis and treatment of abnormal bone growth. PMID- 9858416 TI - Effect of irradiation with a low-intensity diode laser on the metabolism of equine articular cartilage in vitro. AB - OBJECTIVE: To determine whether irradiation with a low-intensity diode laser, which produces radiation at a wavelength of 810 nm, will induce nonthermal enhancement of chondrocyte metabolism. SAMPLE POPULATION: 144 grossly normal articular cartilage explants aseptically harvested from the femoral condyles of 6 adult horses. PROCEDURE: Treated cartilage explants were irradiated with a diode laser at 1 of 7 fluence levels that ranged from 8 to 1,600 J/cm2. Explants were incubated for 24 or 72 hours, labeled for 24 hours with [35S]Na2SO4, and assayed for newly synthesized sulfated glycosaminoglycan (GAG; measured incorporation of 35SO4) and endogenous GAG, chondroitin 6-sulfate (CS), and keratan sulfate (KS) content, using a dimethylmethylene blue assay. Laser-induced temperature changes were measured during irradiation with a diode laser and a neodymium:yttrium aluminum garnet (Nd:YAG) laser, which produces radiation at a wavelength of 1,064 nm, using conditions that were reported in previous studies to increase explant metabolism. RESULTS: After incubation for 24 or 72 hours, rate of 33SO4 uptake or endogenous GAG, CS, or KS content in irradiated explants was not significantly different than in nonirradiated explants. Cartilage temperature increased < 4.75 C during diode laser application. Cartilage temperature increased 5 to 12 C during Nd:YAG laser application. CONCLUSIONS: Minimal thermal increases in cartilage explants with use of a low-intensity diode laser resulted in no change in proteoglycan metabolism of chondrocytes. An increase in tissue temperature over a narrow range with use of a Nd:YAG laser may have contributed to the metabolic alteration of chondrocytes reported in previous studies. PMID- 9858417 TI - Effects of 6alpha-methylprednisolone acetate on an equine osteochondral fragment exercise model. AB - OBJECTIVE: To determine effects of intra-articularly administered 6alpha methylprednisolone acetate (MPA) in exercised horses with carpal osteochondral fragmentation. ANIMALS: 18 horses: 3 groups of 6 each. PROCEDURE: An osteochondral (chip) fragment was created in 1 randomly chosen middle carpal joint of each horse. Polyionic fluid (PF) was injected into both middle carpal joints of horses in the control group. In horses of the MPA-control group, MPA was injected into the middle carpal joint without an osteochondral fragment; a similar volume of PF was injected into the contralateral middle carpal joint. In the MPA-treated group of horses, 100 mg of MPA was injected into the middle carpal joint containing the osteochondral fragment; a similar volume of PF was injected into the contralateral joint. Injections were administered on postsurgical days 14 and 28, and horses were exercised on a high-speed treadmill for 8 weeks, starting on postsurgical day 15. RESULTS: Clinical improvement in degree of lameness was not associated with MPA administration. Joints that contained an osteochondral fragment and were treated with MPA had lower prostaglandin E2 concentration in synovial fluid, and lower scores for intimal hyperplasia and vascularity in synovial membrane, compared with PF-treated joints. However, articular cartilage erosion and morphologic lesions suggested possible deleterious effect of intra-articular MPA administration. CONCLUSIONS: Some beneficial effects of MPA administration on synovial fluid and synovial membrane were identified; however, the deleterious findings contrast with those associated with triamcinolone acetonide used in a similar model, but agree with other results of MPA administration in normal and abnormal joints. PMID- 9858418 TI - Viral excretion in domestic ferrets (Mustela putorius furo) inoculated with a raccoon rabies isolate. AB - OBJECTIVE: To determine susceptibility, incubation and morbidity periods, clinical signs of infection, serologic response, and excretion of virus in domestic ferrets inoculated with rabies virus of raccoon origin. ANIMALS: 54 domestic ferrets. PROCEDURE: 5 groups of ferrets were inoculated IM with the rabies virus. Oral cavity swab specimens and saliva were obtained for virus isolation. Blood was obtained for virus-neutralizing antibody determination. If clinical signs were severe, ferrets were euthanatized immediately. Salivary gland and brain tissue was collected for virus isolation and rabies diagnosis, respectively. RESULTS: Of 51 inoculated ferrets, 19 (37%) were euthanatized with clinical signs of rabies. Mean incubation period was 28 days (range, 17 to 63 days). Clinical signs included ataxia, cachexia, inactivity, paresis, paraparesis, bladder atony, tremors, hypothermia, lethargy, constipation, paralysis, and anorexia. Two rabid ferrets manifested aggressive behavior. Mean morbidity period was 4 to 5 days (range, 1 to 8 days). Virus antigen was detected in brain tissue from all rabid ferrets (n = 19). Two rabid ferrets had detectable virus-neutralizing antibody. Of 32 ferrets that survived, only 1 seroconverted; survivors remained clinically normal throughout the observation period. Rabies virus was isolated from salivary glands of 12 of 19 (63%) rabid ferrets, and 9 (47%) shed virus in saliva. Initiation of virus excretion ranged from 2 days before onset of illness to 6 days after onset. CONCLUSIONS AND CLINICAL RELEVANCE: Rabies should be considered in the differential diagnosis for ferrets that have acute onset of paralysis or behavioral changes and a condition that rapidly deteriorates despite intense medical intervention. PMID- 9858419 TI - "Primary Raynaud's" is not a disease or even a disorder; it's a trait. PMID- 9858420 TI - Primary Raynaud's is a disease or a disorder: it is not merely a trait. PMID- 9858421 TI - The classification of cyclooxygenase inhibitors. AB - In summary, precise classification of COX inhibitors has important clinical implications for efficacy and toxicity. However, classification of these agents clinically is difficult because there are insufficient data to predict correlations between biochemical and pharmacologic properties and the clinical effect of a given agent. In any case, specific COX-2 inhibitors are expected to show antiinflammatory and analgesic activities equivalent to those of NSAID, as well as significant reductions in the incidence of the life threatening side effects (i.e., GI bleeding) associated with COX-1 inhibition. The advantages of preferential COX-2 inhibitors may be more subtle and therefore more difficult to verify in clinical trials. PMID- 9858422 TI - Expression of stem cell factor (SCF) and SCF receptor (c-kit) in synovial membrane in arthritis: correlation with synovial mast cell hyperplasia and inflammation. AB - OBJECTIVE: Stem cell factor (SCF), the ligand for the SCF receptor (c-kit) expressed on precursors and mature mast cells (MC), is a major agonist for human MC (e.g., SCF induces MC development, chemotaxis, activation, proliferation of MC precursors, mediates MC adhesion, and changes MC releasability). We investigated expression of SCF and c-kit in synovial membrane with particular reference to the mechanism of local MC hyperplasia and inflammation in arthritis. METHODS: We conducted single and double labeling immunohistochemistry (ABC, APAAP, indirect immunofluorescence techniques) with antibodies to SCF, c-kit, MC tryptase, Ki-67 antigen (marker for proliferating cells), and CD68 (monocyte/macrophage marker). Synovial specimens analyzed were from 31 patients: traumatic arthritis (TrA, n=9), osteoarthritis (OA, n=12), and rheumatoid arthritis (RA, n=10). Control experiments were performed on human lung, skin, and buccal mucosa tissues, on the HMC-1 mast cell line, and isolated lung MC. Morphometry was performed by computerized image analysis. RESULTS: Synovial c-kit expression was found to be restricted to MC, whereas SCF is detected in synovial lining cells, stromal fibroblasts, monocyte/macrophages, endothelial cells, and in vascular basement membranes. SCF staining was localized to MC as well, but it was not possible to specify whether this represents SCF produced by or bound (via c-kit) to MC. In inflamed synovial membranes/areas, SCF was found to be redistributed into the extracellular matrix. Redistribution of SCF was accompanied by degranulation and/or accumulation of c-kit+ MC, the hyperplasia of which correlated positively with histologic inflammation/inflammatory cell densities, but did not appear to involve MC proliferation in situ. These findings appeared to be common for all the conditions (TrA, OA, RA) studied. CONCLUSION: In addition to the demonstration/characterization of SCF and c-kit protein expression in human synovium, results of this study suggest the hypothesis that, in arthritis, local mobilization of SCF may play a role in the development of synovial MC hyperplasia without inducing in situ proliferation of MC, and that the synovial SCF/MC c-kit system may contribute to the local nonspecific inflammatory response/arthritic flares in TrA, OA, and RA. PMID- 9858423 TI - Ferritin subunits in sera and synovial fluids from patients with rheumatoid arthritis. AB - OBJECTIVE: To determine the proportion of glycosylated ferritin [ferritin bound to concanavalin A (Con-A)] and ferritin subunits in sera and synovial fluids (SF) from patients with rheumatoid arthritis (RA). METHODS: Ferritin concentrations were measured by a sandwich ELISA using rabbit IgG F(ab')2 anti-human ferritin antibody as a coating antibody. Proportions of glycosylated ferritin were examined using Con-A Sepharose 4B. Ferritin subunits were tested by Western blot analysis. RESULTS: Ferritin concentrations in RA SF were significantly elevated compared to those in osteoarthritis (OA) SF (p < 0.01) and those in RA sera (p < 0.01). Percentages of glycosylated ferritin in SF were low in both RA and OA (RA 11.9 +/- 10.7, n = 41; OA 6.9 +/- 11.0, n = 10). However, percentages of glycosylated ferritin in RA sera (65.9 +/- 15.0, n = 20) were significantly higher than in RA SF (p < 0.01). Western blot analysis revealed both G subunit (23 kDa) and L subunit ( 19 kDa) in RA sera, although SF ferritin was composed mostly of L subunit. CONCLUSION: Significant differences in ferritin molecule composition were observed between sera and SF from patients with RA, which suggests that in RA most SF ferritin is synthesized locally in the affected joint. PMID- 9858424 TI - HLA associations of seropositive rheumatoid arthritis in a Cree and Ojibway population. AB - OBJECTIVE: To determine the HLA associations of seropositive rheumatoid arthritis (RA) in a Cree and Ojibway population; to determine whether specific alleles distinguish juvenile or adult onset. METHODS: HLA-A, B, C, and DRB1 alleles were analyzed in 23 Ojibway and Cree patients with RA seen in a single tertiary care center. Comparisons were made with published results of controls and with results of 18 patients with rheumatoid factor (RF) positive polyarticular juvenile rheumatoid arthritis (JRA) from the same population. RESULTS: Comparisons among patients with RA, patients with RF positive polyarticular JRA, and controls showed increased frequencies of the RA shared epitope in patients with RA and of DRB1*0901 in patients with seropositive polyarticular JRA, while the frequency of DRB1*08 alleles was decreased in patients with RF positive polyarticular JRA. CONCLUSION: In this population, DRB1*0901 may promote while DRB1*08 alleles may protect against a juvenile onset of RA specifically. In contrast, the RA shared epitope may have a greater effect on the risk of adult onset seropositive RA. Due to the small patient numbers, these results require confirmation. PMID- 9858425 TI - The performance of the 1987 ARA classification criteria for rheumatoid arthritis in a population based cohort of patients with early inflammatory polyarthritis. American Rheumatism Association. AB - OBJECTIVE: The 1987 American Rheumatism Association (ARA) criteria for rheumatoid arthritis (RA) were developed to discriminate between patients with established RA and those with another rheumatological disorder. Their ability to determine which patients presenting with early synovitis have "true" RA is not known. We evaluated whether the 1987 ARA classification criteria for RA in patients newly presenting with inflammatory polyarthritis (IP) predict persistent, disabling, or erosive arthritis. METHODS: We studied 486 patients with early IP referred to the Norfolk Arthritis Register. The 1987 ARA criteria were applied at baseline, and assessed for their ability to identify (1) patients referred to hospital for whom the diagnosis of RA was recorded by the hospital physician; (2) patients at 3 years with (a) persistent synovitis; (b) moderate or greater disability; and (c) erosions. RESULTS: At baseline, 323 (67%) patients satisfied the ARA criteria in the classification tree format. Exactly 50% of those referred to hospital were given a diagnosis of RA. By 3 years, 76% of the 486 patients had persistent disease, 36% had a Health Assessment Questionnaire score > or = 1, and 40% had erosions. The sensitivity of the criteria was good, ranging from 77 to 87% depending on the outcome. The specificities were poor, and thus the overall discriminatory ability showed little improvement over random probability. CONCLUSION: . Among patients newly presenting with IP, the 1987 ARA criteria for RA had a low ability to discriminate between patients who developed persistent, disabling, or erosive disease and those who did not. Alternative criteria are required for studies investigating early RA. PMID- 9858426 TI - How Canadian and US rheumatologists treat moderate or aggressive rheumatoid arthritis: a survey. AB - OBJECTIVE: To determine which second line agents Canadian and US rheumatologists use to treat patients with active rheumatoid arthritis (RA). METHODS: A one page survey was sent by fax or mail to all 263 members of the Canadian Rheumatology Association and 320 members of the American College of Rheumatology (10% random sample weighted by region) known to practice adult rheumatology. The survey asked for first and second treatment preferences in patients with (1) aggressive RA; (2) moderate RA; and (3) aggressive RA failing a trial of methotrexate (MTX) 25 mg. RESULTS: Altogether 231 (87.8%) Canadian and 230 (71.7%) US rheumatologists responded, and 214 responses in each survey were analyzable. In aggressive RA. MTX was the drug of first choice of most Canadian (68.7%) and US (78.5%) rheumatologists. Intramuscular gold was a drug of first choice for 14.5 and 1.9% of Canadians and Americans, respectively. 93.9% of Canadian and 90.2% of US respondents preferred single agents for the treatment of moderate RA. Among US rheumatologists. no clear leader emerged as a single agent alternative for the management of aggressive RA unresponsive to MTX. Most said they would use combination (38.3%) or triple (23.8%) therapy involving MTX plus sulfasalazine and/or hydroxychloroquine. 52.3% of Canadians preferred single agent therapy, with 34.6% choosing gold as an alternative to MTX. CONCLUSION: Canadian and US rheumatologists preferred MTX for the treatment of aggressive RA. Canadian rheumatologists saw a small but significant role for intramuscular gold. No single agent emerged as a clear alternative to MTX among US rheumatologists. PMID- 9858427 TI - Is bone turnover a determinant of bone mass in rheumatoid arthritis? AB - OBJECTIVE: To study the relationship between bone turnover markers and bone mineral density in patients with rheumatoid arthritis. METHODS: We studied 54 patients, 24 of whom were receiving low dose steroids, and compared them to 54 age and sex matched controls. RESULTS: An 8.2% decrease of femoral neck bone mineral density (BMD) was found in patients not taking steroids compared with controls (confidence interval 1.2-15.3%). Serum markers of bone turnover -- namely, procollagen type I C-terminal propeptide (PICP) and procollagen type I N terminal propeptide (PINP), which reflect bone formation, and procollagen type I C-terminal telopeptide, which reflects bone resorption -- were significantly increased compared with controls (p < 0.05, p < 0.01, p < 0.01, respectively). Both PINP levels and PICP levels were correlated with the femoral neck BMD as well as osteocalcin levels: R = -0.32 (p < 0.05), R = -0.29 (p < 0.05), and R = 0.42 (p < 0.01), respectively. The best independent predictors of bone mass (stepwise multiple regression analysis) at the femoral neck were steroid use, osteocalcin levels, age, height, the presence of rheumatoid factor, and the Health Assessment Questionnaire score, which explained 61.6% of the variance in femoral neck BMD. CONCLUSION: Elderly patients with RA using steroids with severe disease and high levels of osteocalcin have marked osteoporosis at the hip. PMID- 9858429 TI - Dehydroepiandrosterone in systemic lupus erythematosus: relationship between dosage, serum levels, and clinical response. AB - OBJECTIVE: To examine in women with systemic lupus erythematosus (SLE) who participated in a clinical trial the relationship between daily dose of dehydroepiandrosterone (DHEA), serum levels of DHEA and DHEA sulfate (DHEAS), clinical effectiveness, and side effects. METHODS: Twenty-three women with mild to moderate SLE were treated with DHEA for a 6 month period. The starting dose was 50 mg/day, and monthly stepwise increases were allowed. Subjects were assessed monthly by the Systemic Lupus Erythematosus Disease Activity Index, Systemic Lupus Activity Measure (SLAM), Health Assessment Questionnaire, and other outcomes. Serum testosterone, DHEA, and DHEAS levels were obtained and side effects noted monthly. RESULTS: Statistically significant improvements were found in all lupus outcomes over 6 months. Serum DHEA and DHEAS levels correlated with the dose of DHEA. Serum DHEA and DHEAS correlated negatively with SLAM score. A second order regression analysis of serum DHEAS level versus SLAM score suggested that the optimal serum level of DHEAS was 1000 microg/dl. The most common side effect was acne. CONCLUSION: The clinical response to DHEA was not clearly dose dependent. Serum levels of DHEA and DHEAS correlated only weakly with lupus outcomes, but suggested an optimum serum DHEAS of 1000 microg/dl. Monitoring these serum levels appears to have limited clinical utility. PMID- 9858428 TI - Clinical characteristics, treatment, and outcome of adult onset Still's disease in southern Chinese. AB - OBJECTIVE: To study the clinical characteristics, treatment outcome, and complications of patients with adult onset Still's disease (AOSD) in our local Chinese population. METHODS: Patients with AOSD were identified among others who attended our rheumatology clinics from 1967 to 1997 and were followed. Their clinical and laboratory features at presentation, treatment, and outcome were recorded and compared with other reported series. RESULTS: Sixteen patients with AOSD were identified. Eleven (69%) were female. Nine (56%) had onset of the disease between 16 and 35 years of age. The commonest presenting features were fever (100%), arthritis (94%), rash (85%), weight loss (69%), and sore throat (63%). Fifteen patients presented with pyrexia of unknown origin and the median duration of fever before the establishment of the diagnosis was 6 weeks (range 4 75). The acute phase response was marked in all patients with gross elevation of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and complement levels. Hyperferritinemia (> 5 times normal) was present in 90% of cases. Most patients (81%) required corticosteroid therapy and 85% of those steroid treated patients received additional disease modifying agents. The mean duration of followup of our patients was 93.3 months (range 8-362). Five (33%) had monocyclic systemic disease, 6 (40%) had polycyclic systemic disease, and 4 (27%) had frequent relapses that progressed to a chronic arthropathy. CONCLUSION: AOSD in southern Chinese tends to run a benign course, with few patients evolving into chronic inflammatory arthropathy. A significantly lower incidence of serositis, lung involvement, and enlargement of the reticuloendothelial organs was observed at presentation compared with patients of different ethnic origins. PMID- 9858430 TI - Hyperprolactinemia in males with systemic lupus erythematosus. AB - OBJECTIVE: To study prospectively the serum prolactin (PRL) concentrations among male patients with systemic lupus erythematosus (SLE) and their possible relationship to disease activity and manifestations. METHODS: Serum PRL levels were measured by radioimmunoassay in 31 male patients with SLE and 31 age matched controls. Demographic, clinical, and laboratory features of the patients were obtained. Mean PRL levels from both groups were compared, and PRL from patients with SLE was correlated with variables of disease activity, including the SLE Disease Activity Index (SLEDAI), complement level, and anti-dsDNA titer. Thirteen patients were followed serially and changes in PRL levels in relation to fluctuation in disease activity were evaluated. RESULTS: Mean PRL levels were higher in male patients with SLE than healthy controls; however, the difference did not reach statistical significance (230 vs 194 mIU/l; p = 0.06). Hyperprolactinemia was found in 4 patients (13%) and was not associated with particular clinical manifestations or autoantibodies. Considering all patients as a whole, PRL levels did not correlate with variables of disease activity and there was no difference in PRL between patients with active versus inactive disease. A subanalysis of the 4 hyperprolactinemic patients revealed a higher SLEDAI score than those with normal PRL (8.8 vs 3.7; p = 0.20); however, the difference was not statistically significant. Among the hyperprolactinemic patients, PRL levels did not correlate with SLEDAI score or anti-dsDNA titer. Prospective studies of PRL levels in 13 patients did not indicate a role of PRL in the monitoring of disease activity or predicting relapses. CONCLUSION: Hyperprolactinemia occurred in a small proportion of male patients with SLE and its significance remained unclear. Serum PRL level did not correlate with disease activity and was not a reliable marker for disease monitoring. PMID- 9858431 TI - Mycophenolate mofetil suppresses autoimmunity and mortality in the female NZB x NZW F1 mouse model of systemic lupus erythematosus. AB - OBJECTIVE: To examine the effects of the immunosuppressant, mycophenolate mofetil (MM), on autoimmunity, glomerulonephritis, and mortality in the female NZB x NZW F1 (B/W) mouse model of systemic lupus erythematosus (SLE). METHODS: The development of murine lupus was assessed during the lifespan of 10 female B/W mice given 200 mg/kg/day of MM compared to 10 female B/W mice given vehicle. At 6 week intervals, mice were examined for weight change, albuminuria, antibodies to DNA, and IgG immunoglobulin levels. Morbidity and mortality were assessed daily. In a parallel study, MM treated and control B/W mice were examined at 18 weeks of age for splenocyte phenotype and adhesion molecule expression, as well as antibody titers and in vitro cytokine production in response to immunization with dinitrophenyl-keyhole limpet hemocyanin (DNP-KLH). RESULTS: The administration of MM was well tolerated without apparent side effects. Weight gain in MM treated and control mice was identical through 36 weeks of age. In the treatment group, MM suppressed the development of albuminuria and anti-DNA antibodies compared to the control animals. There were no significant differences between groups in serum concentrations of total IgG. At 60 weeks of age survival in the MM treated group was 100% compared to 10% in the control group. MM did not alter the percentages of CD4, CD8, or IgM positive splenocytes; however, the percentage of CD4+ T lymphocytes expressing very late antigen 4 and intercellular adhesion molecule 1 was reduced. MM inhibited the antibody response to DNP-KLH immunization in vivo; however, in vitro cytokine production in response to KLH was not suppressed. CONCLUSION: MM suppressed the development of autoimmunity and prolonged lifespan in the female B/W mouse model of SLE. Suppression of autoimmunity was achieved without obvious side effects or altered CD4:CD8 T cell ratios. MM may be a useful primary or adjunctive therapy in human SLE. PMID- 9858432 TI - Excessive synthesis of matrix metalloproteinases in exocrine tissues of NOD mouse models for Sjogren's syndrome. AB - OBJECTIVE: Matrix metalloproteinases (MMP) and their substrates, components of the extracellular matrix, regulate environmental signals for cellular differentiation and tissue function. Changes in the levels of these enzymes may influence cell survival as well as pathology involving ectopic apoptosis. Using the non-obese diabetic (NOD) mouse model for Sjogren's syndrome, we evaluated the synthesis and expression of MMP in the exocrine target tissues of autoimmunity. METHODS: NOD, immunodeficient NOD-scid, and nondiabetic NOD.B10.H2b mice were evaluated for MMP activity in their saliva and exocrine gland lysates by gelatin zymography and reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, changes in protein content of saliva and gland lysates were determined by specific Western blot and by enzymatic activity of amylase and cysteine proteases. Mice continuously treated with the MMP inhibitor GM6001 were evaluated from 7 to 20 weeks of age for the contribution of MMP activity to development of these hallmark biochemical markers of Sjogren's syndrome-like disease of NOD mice. RESULTS: Gelatin zymography of whole saliva and gland lysates indicated the presence of increased proteolytic activity, corresponding to proteins with a molecular mass ranging from 50 to 95 kDa, in the saliva of older (> 20 weeks of age) NOD mice as well as NOD.B10.H2b and NOD-scid mice compared to BALB/c controls. Elevated steady state levels of mRNA transcripts for the gelatinases MMP-2 and MMP-9 were detected in total RNA extracted from parotid and submandibular glands by RT-PCR. Despite prophylactic injection of the broad spectrum MMP inhibitor GM6001 into mice beginning at 7 weeks of age and continuing to 20 weeks, development of the autoimmune exocrinopathy was neither stopped nor retarded. CONCLUSION: These observations suggest that excessive MMP activity is associated with autoimmune Sjogren's syndrome-like disease in NOD mice. However, a possible contribution by increased MMP activity in initiation and progression of this autoimmune disease is yet to be elucidated. PMID- 9858433 TI - Kininogen-kallikrein-kinin system in plasma and saliva of patients with Sjogren's syndrome. AB - OBJECTIVE: To evaluate variables of the kininogen-kallikrein-kinin system (KKKS) simultaneously in plasma and saliva of patients with Sjogren's syndrome (SS). METHODS: We studied a group of 20 female patients with SS aged 37-75 years, 7 with primary SS (SS1) and 13 with SS secondary to rheumatoid arthritis (SS2), and 20 healthy individuals. Total kininogen and high and low molecular weight kininogen (HKg and LKg, respectively) levels were evaluated by ELISA. The activity of plasma and tissue kallikreins was determined by enzyme activity on selective chromogenic substrates. RESULTS: The plasma levels of total kininogen, HKg, and LKg, and the activity of plasma kallikrein observed in patients were not significantly different from controls. The tissue kallikrein-like activity in plasma and the active tissue kallikrein in saliva were significantly increased in patients with SS, whereas the total salivary tissue kallikrein activity in patients was not significantly different from controls. The concentration of protein in the saliva of patients was significantly increased, and a positive correlation between salivary protein levels and the active tissue kallikrein was observed. CONCLUSION: Comparisons between the total and the active tissue kallikrein in saliva of patients with SS showed that most of the tissue kallikrein was in its active form. In addition, we observed a concomitant increase of the tissue kallikrein-like activity in plasma. These results suggest increased activation of the KKKS in plasma and saliva of patients with SS. PMID- 9858434 TI - Autonomic cardiovascular neuropathy in Sjogren's syndrome. A controlled study. AB - OBJECTIVE: To test patients with primary Sjogren's syndrome (SS) for evidence of autonomic neuropathy. METHODS: Thirty-two patients with primary SS and 22 age and sex matched healthy individuals were asked specific questions about symptoms suggestive of autonomic neuropathy, and were subjected to a battery of 5 cardiovascular tests: response of blood pressure to sustained hand grip, Valsalva maneuver, heart rate response to deep breathing, and heart rate and blood pressure response to standing up. The chi-squared test with Yates' correction and 95% confidence intervals were used for statistical analysis of the results. RESULTS: Sixteen patients (50%) had symptoms of autonomic neuropathy when specifically asked versus none of the controls (p < 0.0005). The frequency of abnormal responses to the tests was 68.8% in patients and 12.7% in controls (p < 0.0001). Severe autonomic cardiovascular neuropathy was found in 87.5% of the patients but in none of the healthy individuals (p < 0.0001). CONCLUSION: Our results suggest that autonomic neuropathy is a feature of a significant portion of the SS population, and such patients should have appropriate evaluation. Similarly, patients with unexplained autonomic neuropathy should be investigated for evidence of SS. PMID- 9858436 TI - Radiographic diagnosis of sacroiliitis--are sacroiliac views really better? AB - OBJECTIVE: To determine whether detailed oblique radiographs of the sacroiliac (SI) joints provide significant diagnostic advantage to a single AP projection of the pelvis in establishing the presence and severity of sacroiliitis. METHODS: Radiographs (both AP pelvis and detailed oblique projections) of 445 right SI joints and 442 left SI joints were obtained from patients with an established diagnosis of a seronegative spondyloarthropathy, and interpreted for severity. Data for the right and left SI joints were analyzed, comparing interpretations of severity based on AP pelvis projections with oblique views. RESULTS: Analysis of these data showed an agreement rate between AP views and SI views of 89.7% for the right SI joint radiographs and 86.4% for the left SI joint. There was no instance in which a patient with "unequivocal abnormalities"of the SI joints on the AP pelvis was read as having "normal" SI views. Similarly, there were no cases in which "normal" SI joints on AP pelvis films were read as having unequivocal abnormalities on SI views. CONCLUSION: In this group of patients with seronegative spondyloarthropathies, there was very close agreement between severity score of sacroiliitis from AP pelvis radiographs and SI joint views. We conclude that in most circumstances, the AP pelvis film will yield the diagnosis of sacroiliitis without the additional radiation exposure and expense related to specific SI joint radiographs. PMID- 9858435 TI - Serum testosterone levels are not elevated in patients with ankylosing spondylitis. AB - OBJECTIVE: Studies in patients with ankylosing spondylitis (AS) describe slightly elevated serum testosterone levels, but these studies were not properly controlled for possible confounders. METHODS: In a case-control study serum levels of sex steroids, luteinizing hormone, and sex hormone binding globulin (SHGB) were measured in patients with AS and in age and sex matched controls. The body mass index, smoking status, use of alcohol, and fat intake were recorded. RESULTS: Testosterone levels measured in serum extracts did not differ in 50 male patients with AS compared to controls (mean +/- SD 16 +/- 4 vs 15 +/- 5 nmol/l, respectively; p = 0.54). In unextracted serum, however, male patients showed elevated testosterone (p < 0.001) and dehydroepiandrosterone sulfate levels (p = 0.003), even after controlling for confounders (p < 0.001). One of 10 female patients had an elevated testosterone level in unextracted serum. The 17 male users and one of the 2 female users of phenylbutazone had the highest testosterone levels in unextracted serum, and all showed a significant decline after extraction. Serum levels of other sex steroids, luteinizing hormone, and SHGB did not differ significantly between patients and controls. CONCLUSION: Serum testosterone levels are not elevated in male patients with AS. Spuriously elevated testosterone levels in unextracted serum might be related to the use of phenylbutazone in our patient sample. PMID- 9858437 TI - Chronic arthritis and carpo:metacarpal ratio in Japanese patients with adult Still's disease. AB - OBJECTIVE: To characterize Japanese patients having adult Still's disease (ASD) with chronic arthritis (> 6 months) and to examine the association of chronic arthritis with carpo:metacarpal ratio (CMC ratio), an index of radiographic progression in rheumatoid arthritis (RA). METHODS: Twenty-seven patients with ASD (16 women and 11 men, mean age at disease onset 27.7 years) were classified into 2 groups: patients with (chronic articular ASD, 16 patients, 59%) or without (systemic ASD, 11 patients, 41%) chronic arthritis. Clinical and laboratory findings were compared between both groups. CMC ratio was calculated on serial hand radiographs in patients with chronic articular ASD. RESULTS: In our series, serositis was rarely observed in chronic articular ASD. Peripheral arthritis (including transient arthritis), such as metacarpophalangeal, proximal interphalangeal, or ankle joint, was more frequently observed in chronic articular ASD than in systemic ASD (p < 0.05). Wrist arthritis was frequently observed also in systemic ASD; however, joint space narrowing of carpometacarpal or intercarpal joints was recognized only in chronic articular ASD (44%). CMC ratio at the last observation in 14 patients with chronic articular ASD was significantly decreased (0.526 +/- 0.039) compared to that at disease onset (0.553 +/- 0.034) (p < 0.05), while no decrease was observed in 4 with systemic ASD (0.565 +/- 0.062 at disease onset, 0.563 +/- 0.043 at the last observation). CONCLUSION: It is suggested that chronic articular ASD has certain characteristics. CMC ratio may be a quantitative index for assessment of radiographic changes of carpal joints, not only in RA but also in chronic articular ASD. PMID- 9858438 TI - Tumor necrosis factor-alpha is not essential in endotoxin induced eye inflammation: studies in cytokine receptor deficient mice. AB - OBJECTIVE: Anterior uveitis frequently occurs in association with specific systemic inflammatory diseases. Interleukin 1 (IL-1) and tumor necrosis factor alpha (TNF-alpha) have been implicated in the pathogenesis of these diseases. We evaluate the need for these cytokines in a model of anterior uveitis. METHODS: Endotoxin was injected into the vitreous of mice deficient in IL-1 receptor type I, TNF receptors p55 and p75, both IL-1R1 and TNFR p55, or controls. Eyes were harvested after 24 h for histology and IL-6 bioassays or after 3 h for reverse transcriptase-polymerase chain reaction analysis of mRNA for specific cytokines or enzymes. RESULTS: No significant difference in the number of infiltrating cells was found in TNFR p55/p75 deficient mice compared to controls in any of 4 separate experiments or in the combined data (p = 0.8). The number of infiltrating cells was significantly reduced in 2 of 4 experiments with IL-1R1 deficient mice (p < 0.001 based on combined data from 4 studies). IL-1R1/TNFR p55 deficient mice had a reduction in infiltrating cells in 2 of 3 experiments (p < 0.001 based on combined data from all studies). IL-6 levels were not significantly reduced in either of 2 experiments with TNFR p55/p75 deficient mice, but were reduced in one of 2 experiments with IL-1R1-/- mice (p = 0.02) and in one experiment with IL-1R1/TNFR p55 deficient mice (p = 0.01). In response to endotoxin, all 3 receptor deficient lines increased mRNA levels for IL-1-alpha, IL-10, TNF-alpha, IL-1 receptor antagonist, and inducible nitric oxide synthase. CONCLUSIONS: IL-1 appears to have a more pivotal role in endotoxin induced uveitis than TNF-alpha, although neither cytokine is essential. Deletion of receptors for both cytokines has the most consistent effect, which is in accord with the hypothesis that these cytokines are, at least in part, functionally redundant. PMID- 9858439 TI - Diacerhein and rhein reduce the interleukin 1beta stimulated inducible nitric oxide synthesis level and activity while stimulating cyclooxygenase-2 synthesis in human osteoarthritic chondrocytes. AB - OBJECTIVE: To evaluate the in vitro effects of diacerhein, a new drug for the treatment of osteoarthritis (OA), and its active metabolite, rhein, on the production of nitric oxide (NO), prostaglandin (PGE2), cyclooxygenase-2 (COX-2), as well as the production and expression of the inducible nitric oxide synthase (iNOS) in human OA chondrocytes. These results were compared to those of the nonsteroidal antiinflammatory drug (NSAID) naproxen. METHODS: Human OA chondrocytes were incubated in the presence or absence of 25 units/ml recombinant human interleukin-1beta (rhIL-1beta) with or without therapeutic concentrations of diacerhein and rhein at 5, 10, and 20 microg/ml and naproxen at 30 and 90 microg/ml. Effect of the drugs was also tested on both OA chondrocytes and cartilage explants on increasing IL-1beta concentration (0-100 units/ml). The NO and PGE2 levels were determined in the culture medium using the Griess reaction and a specific ELISA, respectively. Production of COX-2 and synthesis and expression of iNOS were quantitated by Western blot and Northern blot, respectively. RESULTS: The IL- 1beta induced NO production was inhibited by both diacerhein and rhein in a time and dose dependent fashion, with statistical significance reached at the therapeutic concentration of 20 microg/ml. A decrease over 80% was found at 24, 48, and 72 h incubation. This was consistent for both chondrocytes and cartilage explants even in the presence of high IL-1beta concentration (100 units/ml). Moreover, this effect appeared to result from iNOS transcriptional and/or post-transcriptional events as indicated by a decrease in this enzyme level for both the mRNA and protein. Naproxen, however, showed only a slight inhibition of IL-1beta induced NO production at the highest dose used, 90 microg/ml. A maximum decrease of 23% in IL-1beta induced NO production was recorded after a 72 h incubation. In contrast to naproxen, which abrogated PGE2 and had no effect on COX-2 synthesis, rhein and diacerhein at 5 and 10 microg/ml produced an enhancement in their levels. CONCLUSION: Diacerhein and rhein, in contrast to an NSAID, are potent inhibitors of IL-1beta induced NO production by chondrocytes and cartilage, without reducing PGE2 production. PMID- 9858440 TI - Piroxicam therapy: a double blind, randomized, multicenter study comparing 2 versus 4 week treatment in patients with painful knee osteoarthritis with effusion. AB - OBJECTIVE: To compare the efficacy and safety of piroxicam 20 mg once a day for 14 or 28 days in patients with knee osteoarthritis (OA) and synovial effusion. METHODS: We conducted a multicenter, randomized, double blind study in 1905 outpatients. Efficacy was assessed by changes in synovial effusion, pain on a 100 mm visual analog scale (VAS), and impairment using Lequesne's functional index. Patients were classified at Day 28 as improved (defined as VAS and Lequesne index decrease of at least 30% from Day 14), worsened (defined as VAS and Lequesne index increase of at least 30% from Day 14), or unchanged. Safety was assessed on the basis of adverse events reported by the patients. RESULTS: After 14 days, changes in pain, synovial effusion, and functional impairment significantly decreased from baseline within each group (p < 0.001, respectively), but did not differ between the groups. Between 14 and 28 days, outcome measure changes were significantly better in the 28 day group, p = 0.01, 0.0001, and 0.0001, respectively. In the 28 day and 14 day groups, improvement with regard to pain was observed for 339 (52.4%) and 280 (29.4%) patients, respectively, (p < 0.0001), and with regard to functional impairment for 298 (31.5%) and 233 (24.3%) patients (p < 0.0001). Adverse events accounted for 7.5 and 6.7% of withdrawals in the 28 day and 14 day groups, respectively. CONCLUSION: When administration of piroxicam 20 mg is prolonged to 28 days, continuing benefit is observed for some patients with knee OA with painful synovial effusion without a significant difference in safety. PMID- 9858441 TI - The effectiveness of exercise therapy in patients with osteoarthritis of the hip or knee: a randomized clinical trial. AB - OBJECTIVE: To determine the effectiveness of exercise therapy in patients with osteoarthritis (OA) of the hip or knee. METHODS: A randomized single blind, clinical trial was conducted in a primary care setting. Patients with hip or knee OA by American College of Rheumatology criteria were selected. Two intervention groups were compared. Both groups received treatment from the patients' general practitioner, including patient education and medication if necessary. The experimental group also received exercise therapy from a physiotherapist in primary care. The treatment period was 12 weeks. The main outcome measures were pain, medication use (nonsteroidal antiinflammatory drugs, NSAID) and observed disability. RESULTS: A total of 201 patients were randomized. Exercise therapy was associated with a reduction of pain in the past week (difference in change 17.0; 95% CI -23.6, -10.4) and observed disability (-0.19; 95% CI -0.38; -0.01). Effect sizes were medium (0.58) and small (0.28), respectively. No effect of exercise therapy was found for the use of NSAID. Additional beneficial effects (p = 0.05) were found for the use of paracetamol (effect size 0.33), global effect as perceived by the patient (effect size 0.68), and muscle strength of the hip (effect size 0.34). CONCLUSION: After 12 weeks, exercise therapy is effective in reducing pain and disability. The size of the effects is medium and small, respectively. PMID- 9858442 TI - Pentosidine in synovial fluid in osteoarthritis and rheumatoid arthritis: relationship with disease activity in rheumatoid arthritis. AB - OBJECTIVE: Pentosidine is an advanced glycation endproduct formed by glycosylation and oxidation. Our aim was to develop a means to measure pentosidine in synovial fluid (SF), and to compare its concentration in SF in patients with osteoarthritis (OA) and rheumatoid arthritis (RA), and to investigate the relationship between its concentration in SF and the disease activity of RA. METHODS: SF was collected from knee joints in 31 patients with RA and 40 with OA, who had hydrarthrosis. One patient with RA and 7 with OA who had the complication of diabetes mellitus or chronic renal failure made up the DM/CRF group, and the remaining patients made up the RA group (n = 30) and the OA group (n = 33). Pentosidine was measured by the direct HPLC method with column switching after hydrolysis of SF. RESULTS: Pentosidine was detected in all SF and was greater in RA (83.9 +/- 46.0 nmol/l, mean +/- SD) than in OA (40.1 +/- 19.6 nmol/l). Three DM/CRF patients undergoing hemodialysis had markedly high pentosidine levels (482.5 +/- 280.8 nmol/l). There was a significant correlation between pentosidine and C-reactive protein (CRP), erythrocyte sedimentation rate, and Lansbury Index (p < 0.01). Patients with RA were divided into high and low activity groups according to the CRP and Lansbury Index. Pentosidine was significantly higher in the high activity group (CRP > or = 2.0 mg/dl and Lansbury Index > or = 50%) than in the low activity group (CRP < 2.0 and/or Lansbury Index < 50) (100.9 +/- 42.8 vs 58.5 +/- 39.6 nmol/; p = 0.0013). CONCLUSION: Pentosidine in synovial fluid was higher in RA than in OA. Pentosidine levels in SF were related to the disease activity in RA. PMID- 9858443 TI - Prevalence of juvenile chronic arthritis and familial Mediterranean fever in Turkey: a field study. AB - OBJECTIVE: To investigate the prevalence of juvenile chronic arthritis (JCA), familial Mediterranean fever (FMF), and Behcet's disease in Turkish children through a field survey. METHODS: The field survey was based on cluster centering with 2 level strata. A total of 46,813 children were screened. For the diagnosis of chronic arthritis and Behcet's previously suggested criteria were used. We have developed criteria for the diagnosis of probable FMF. Children previously diagnosed to have these diseases were also defined and included. RESULTS: JCA was found in 6.4/10,000. 2.8/10,000 children were previously diagnosed as FMF (minimum phenotype frequency). Together with the probable diagnosis of FMF, the prevalence increased to 9.3/10,000. The findings were also compared with those of our center. None of the 46,813 children had Behcet's disease. CONCLUSION: The prevalence of chronic arthritis is similar to the other childhood populations reported. However, FMF has a very high prevalence. PMID- 9858445 TI - Improvement of systemic lupus erythematosus activity by the association of delayed onset Cushing's syndrome. AB - We describe a 43-year-old woman with systemic lupus erythematosus (SLE) who had complete resolution of all SLE symptoms as a result of hypercortisolemia induced by a glucocorticoid-producing adenoma of the left adrenal gland. After an adrenalectomy, she developed an SLE exacerbation, characterized by photosensitivity, polyarthralgia, and hemolytic anemia, which required intensive steroid therapy. This is the first report of a patient with SLE entering apparent remission due to excessive adrenal secretion of glucocorticoids. PMID- 9858444 TI - Development of bone mineral density at the lumbar spine and femoral neck in juvenile chronic arthritis--a prospective one year followup study. AB - OBJECTIVE: To determine the magnitude of lumbar and femoral bone mineral gain in patients with juvenile chronic arthritis (JCA) using dual X-ray absorptiometry. METHODS: Bone mineral density (BMD) was measured at entry and again after 12 months at the lumbar spine and femoral neck in healthy children (n = 65) and children with oligoarticular (n = 36) and polyarticular (n = 69) JCA. Five of the oligoarticular and 38 polyarticular patients were treated with systemic glucocorticoids. In addition to the changes in BMD, the annual changes in calculated bone mineral volumetric density (BMDvol) and bone size were determined simultaneously. RESULTS: In polyarticular JCA, the acquisition of BMD was decreased at the femoral neck (2.2 vs 4.8%; p < 0.05), but remained the same at the spine compared with healthy children; in oligoarticular JCA, the increase in BMD at the femoral neck was similar to that in controls, but significantly increased at the spine compared with the change in the control group (7.4 vs 4.9%; p < 0.05). The detected annual changes in BMD were associated with the changes in BMDvol. Bone mineral gain was significantly delayed at the lumbar spine in children treated with glucocorticoids. CONCLUSION: In children with JCA, the development of bone mineral is different at the lumbar spine and at the femoral neck, but it also depends on the subtype of JCA and on the use of systemic glucocorticoids. PMID- 9858446 TI - Diagnostic confusion caused by hepatitis C: hemochromatosis presenting as rheumatoid arthritis. AB - We describe a patient with hemochromatosis and coexistent infection with the hepatitis C virus who was initially thought to have rheumatoid arthritis. His symptoms began at the age of 44 with pain of the hand joints, shoulders, hips, and knees and a positive rheumatoid factor. Four years later, he required replacement of both hips due to severe hip arthritis. Abnormalities in liver function were noted early on, but they were attributed to infection with the hepatitis C virus, detected serologically and by polymerase chain reaction amplification in the blood. The correct diagnosis was delayed until a decision to use methotrexate as treatment for his arthritis led to a liver biopsy, which revealed increased iron deposition consistent with hemochromatosis, confirmed by genetic testing, which revealed that the patient was homozygous for the C282Y mutation of the HLA-H gene. PMID- 9858447 TI - Multiple cerebellar infarction due to vertebral artery obstruction and bulbar symptoms associated with vertical subluxation and atlanto-occipital subluxation in ankylosing spondylitis. AB - Ankylosing spondylitis (AS) results in disease-specific inflammation at the site of ligamentous insertion into the bone. Atlantoaxial joint subluxation and vertical subluxation of the axis may occur as a consequence of instability resulting from the inflammatory process. Spontaneous anterior atlantoaxial subluxation is a well recognized complication in about 2% of patients with AS, and presents with or without signs of spinal cord compression. Vertical subluxation may follow anterior or posterior subluxation. It was noted in 3-8% of patients with rheumatoid arthritis, but is an exceedingly rare complication of AS. Moreover, it has never been reported that multiple cerebellar infarction and bulbar symptoms developed spontaneously due to atlanto-occipital subluxation and vertical subluxation in a patient with a long [corrected] history of AS. We describe a man with AS who developed multiple cerebellar infarction due to vertebral artery obstruction and bulbar symptoms associated with atlanto occipital subluxation and vertical subluxation. PMID- 9858448 TI - Behcet's disease with unusual cutaneous lesions. AB - A variety of cutaneous lesions may occur in Behcet's disease (BD) both at presentation and over the course of the disease. Skin involvement of the hands and feet has been infrequently observed. We describe a patient with BD with recurrent, multiple, papulonodular cutaneous lesions affecting the palm and fingers of both hands, occurring simultaneously with aphthous stomatitis. The lesions consisted of roundish, erythematous, painful, bluish-red nodules, 0.5-1 cm in diameter, with a "pernio-like" aspect. Histologic examination revealed perivascular neutrophilic infiltrates. We suggest that cutaneous lesions with a pernio-like aspect as observed in our patient may be included in the spectrum of the cutaneous manifestations of Behcet's disease. PMID- 9858449 TI - Folic acid supplementation prevents deficient blood. PMID- 9858450 TI - Managed care and the practice of rheumatology. PMID- 9858451 TI - Additional surgery after breast device implantation. PMID- 9858452 TI - Adjustment in patients with rheumatoid arthritis and their children. PMID- 9858453 TI - What use are fibromyalgia control points? PMID- 9858454 TI - Sudden sensorineural hearing loss in patients with systemic lupus erythematosus or lupus-like syndrome and antiphospholipid antibodies. PMID- 9858455 TI - Anti-annexin V antibodies and osteonecrosis in systemic lupus erythematosus. PMID- 9858456 TI - Controlling hyperuricemia. PMID- 9858457 TI - Multifocal osteonecrosis after chemotherapy in a patient with breast cancer. PMID- 9858458 TI - Sp1 screening test in risk assessment for osteoporosis. PMID- 9858459 TI - Soluble tumor necrosis factor receptor levels in familial Mediterranean fever. PMID- 9858460 TI - Phylogenetic analysis of the genus Bifidobacterium and related genera based on 16S rDNA sequences. AB - The 16S rRNA gene sequences were determined for type strains of 21 Bifidobacterium species. A phylogenetic tree was constructed using the determined sequences and sequences from DNA databases, which contain the sequences of 11 type strains of Bifidobacterium species and 11 strains of related genera. All species of the genus Bifidobacterium and Gardnerella vaginalis ATCC 14018 belonged to a cluster phylogenetically distinct from the other genera. The cluster was divided into two subclusters: subcluster 1 composed of most species of Bifidobacterium and G. vaginalis, and subcluster 2 consisting of two species, B. denticolens and B. inopinatum; both of which were isolated from human dental caries. In the genus Bifidobacterium, four groups of species are known to be moderately to highly related by DNA-DNA hybridization. The four groups of species exhibited more than 99% similarity among their 16S rDNA sequences within each group. These results indicated that species with around 99% or more similarity in their 16S rDNA sequences should be confirmed for species identities. PMID- 9858461 TI - Serological studies of an acid-labile O-polysaccharide of Proteus vulgaris OX19 lipopolysaccharide using human and rabbit antibodies. AB - In a Weil-Felix test, sera from patients infected with Rickettsia sp. agglutinate Proteus OX types of bacteria and Proteus lipopolysaccharide (LPS) are responsible for the cross-reaction. Data on the character of LPS of one of the OX group strains, Proteus vulgaris OX19, are contradictory, and it remained unclear whether it has an O-polysaccharide (OPS) and is thus LPS of the smooth type (S) or not (rough-type LPS). Our studies showed that P. vulgaris OX19 (strain PZH-24) produces a smooth-type LPS that contains a long-chain OPS, but it undergoes depolymerization during mild acid hydrolysis conventionally used for LPS delipidation and loses the serological activity. An elucidation of the complete structure of OPS demonstrated the presence of a glycosyl phosphate linkage responsible for the acid-lability of the polysaccharide chain. In ELISA, both IgM type antibodies in a Weil-Felix test with human anti-Rickettsia typhi sera and rabbit anti-P. vulgaris OX19 antibodies reacted with OPS. Rabbit antibodies did not inhibit the cross-reaction with human antibodies and thus bind to different epitopes. PMID- 9858462 TI - Analysis of genetic relationships and antimicrobial susceptibility of verotoxin producing Escherichia coli strains isolated in Nagasaki Prefecture, Japan in 1996. AB - A total of 19 Escherichia coli O157 isolates were obtained in Nagasaki Prefecture, in the south-western part of Japan, between 1990 and 1996. Pulsed field gel electrophoresis (PFGE) and computer-assisted analysis were applied to determine genetic relationships among these strains. Fragment patterns of the isolates in Nagasaki, as determined by PFGE, were compared with those of isolates in other areas where large outbreaks and sporadic cases of E. coli O157 infection occurred. Similarity values of all the strains isolated in Nagasaki Prefecture were over 0.65 except for E. coli O26. Some strains were identical to the strains isolated from the areas where large outbreaks occurred. All strains were susceptible to ampicillin, fosfomycin, minocycline, amikacin, ofloxacin and sulfamethoxazole-trimethoprim. PMID- 9858463 TI - Detection and long-term existence of Shiga toxin (Stx)-producing Escherichia coli in sheep. AB - The isolation and characterization of Shiga-like toxin (Stx)-producing Escherichia coli (STEC) from sheep are described. The distribution of stx genes in E. coli isolates was detected by PCR. When brain heart infusion (BHI) broth and novobiocin supplemented m-EC broth (N-mEC) were used as enrichment culture for the isolation of STEC, N-mEC, compared to BHI, showed clearly lower efficiency. Finally, 5 STEC isolates from 4 sheep were isolated and characterized by biochemical and genetical analysis. All of them were confirmed by ELISA and Vero cell cytotoxicity assay for the production of Stx. Moreover, some strains carried hemolysin and eaeA genes and harbored large plasmids. Based on their plasmid profiles, antibiotic patterns and PCR-based DNA fingerprinting analysis using random amplified polymorphic DNA (RAPD), all isolates were different from each other. Three of the isolates were identified to belong to serogroups O2, O153 and O165, respectively, and the STEC strains belonging to these serogroups had been isolated from STEC outbreaks in humans. Four months after the first isolation in July 1997, STEC from sheep #1 was isolated again. A new isolate, HI 11, was identified as STEC O2:Hnt. Simultaneously, 2 STEC, which were genetically and phenotypically different from each other, were isolated from the same sheep at intervals of 4 months. These results demonstrate that sheep may be an important animal for studying human STEC infections, and that further epidemiological surveys on STEC are necessary. PMID- 9858464 TI - Rapid serodiagnosis of Mycobacterium avium-intracellulare complex infection by ELISA with cord factor (trehalose 6, 6'-dimycolate), and serotyping using the glycopeptidolipid antigen. AB - Mycobacterium avium-intracellulare complex (MAC) is one of the most important opportunistic pathogens, particularly in patients with acquired immunodeficiency syndrome (AIDS). The aim of this study was to determine whether an enzyme-linked immunosorbent assay (ELISA) using trehalose 6,6'-dimycolate (TDM) as an antigen can be used for the rapid serodiagnosis of MAC infection. We also identified MAC serotypes by ELISA using serotype-specific glycopeptidolipid (GPL) antigen. To confirm our findings, the thin-layer chromatographic (TLC) behavior of serotype specific GPL of the strains isolated from MAC-infected patients was also tested. Forty patients infected with MAC and 30 healthy controls were tested. Thirty-two of the 40 MAC-infected patients had higher titers of serum antibodies against MAC TDM than against MTB TDM, while all 30 healthy control sera were unreactive to MAC TDM and MTB TDM. Results of the GPL ELISA indicated that 20 of the 40 MAC infected patients' sera were reactive against serotype 4 GPL, 3 against serotype 8 GPL, and 1 against serotype 16 GPL. A TLC analysis of the GPL of the 40 MAC isolates showed that 16 strains were of serotype 4, 5 of serotype 8, and 2 of serotype 16. Results of the GPL ELISA were in good accord with those of the TLC analysis for most patients. Our findings suggest that ELISA using TDM is useful for rapid serodiagnosis of MAC infection, and that complementary ELISA testing using serotype-specific GPL gives additional detailed information concerning MAC serotypes. PMID- 9858465 TI - Attachment of nontypable Haemophilus influenzae to human pharyngeal epithelial cells mediated by a ganglioside receptor. AB - Nontypable Haemophilus influenzae (NTHI) is one of the major pathogens of human respiratory infections and has the ability to attach to pharyngeal epithelial cells. We characterized the epithelial cell receptor to which NTHI bind. Neuraminidase pretreatment of pharyngeal epithelial cells resulted in a significant decrease in NTHI attachment, suggesting sialic acid as an important component of the receptor. The attachment was not decreased in NTHI pretreated with 1,000 microg/ml of fucose, N-acetyl-neuraminic acid, N-acetyl-glucosamine, N acetyl-galactosamine, acetyl-salicylic acid and colominic acid. Only treatment with gangliosides D1a, D1b and D2 at a concentration of 12.5 microg/ml significantly decreased the attachment. On the other hand, treatment with gangliosides M1, M2, M3, D3, T1b and asialoganglioside M1 did not decrease the attachment of NTHI. Only ganglioside D2 inhibited the attachment significantly at a concentration of 12.5 ng/ml. Other isolates of NTHI showed a decrease in attachment after treatment with ganglioside D2. Treatment of cells with anti human GD2 monoclonal antibody also decreased the attachment of NTHI in a dose dependent manner. This study indicates that sialic acid glycoconjugate, GD2, is one of the receptors of NTHI on human pharyngeal epithelial cells. PMID- 9858466 TI - Purification and characterization of enterotoxin produced by Aeromonas sobria. AB - We purified the toxin of Aeromonas sobria capable of inducing a positive response in the mouse intestinal loop assay. The purified toxin showed a positive response not only in the loop assay but also in a hemolytic assay. Subsequently, we cloned the toxin gene and demonstrated that the product of this gene possessed both hemolytic and enterotoxic activities. These results showed that the enterotoxin of A. sobria possesses hemolytic activity. Nucleotide sequence determination of the toxin gene and amino acid sequence analysis of the purified toxin revealed that it is synthesized as a precursor composed of 488 amino acid residues, and that the 24 amino-terminal amino acid residues of the precursor is removed in the mature toxin. As antiserum against the purified toxin neutralized the fluid accumulation induced by living cells not only of A. sobria but also of A. hydrophila, this and antigenically related toxin(s) are thought to play an essential role in the induction of diarrhea by these organisms. The toxin-injured Chinese hamster ovary (CHO) cells induced the release of intracellular lactose dehydrogenase (LDH). The release of LDH from CHO cells and the lysis of erythrocytes by the toxin were repressed by the addition of dextran to the reaction solution, indicating that the toxin forms pores in the membranes and that the cells were injured by the osmotic gradient developed due to pore formation. However, the histopathological examination of intestinal cells exposed to the toxin showed that it caused fluid accumulation in the mouse intestinal loop without causing cellular damage. PMID- 9858467 TI - Genomic analysis of the viral population in genital secretions early after infection of simian immunodeficiency viruses in macaque monkeys. AB - To clarify the change in the viral population during passage from the vaginal cavity to blood circulation and vice versa, we examined the viral clones detected in cells in vaginal washes (VWCs) early after inoculation and after systemic infection with polyclonal SIV. In two intravaginally inoculated monkeys, the viral clones found in VWCs at 18 days p.i. were shown to be some of those contained in the inoculum, whereas the viral population in the peripheral blood mononuclear cells (PBMCs) was a monotype. This gradual decrease of viral clones suggested the possible existence of two barriers, one at the genital tract and the other between the genital tract and the blood. Later, at one month p.i., the viral clones in VWCs became rather restricted, whereas those in PBMCs diverged from a single clone to several clones. This suggested that different mechanisms affect the viral populations in PBMCs and VWCs. In order to examine how the viral population was affected by passage from the blood to the vaginal cavity, a monkey was intravenously inoculated and the viral clones in VWCs were analyzed at 14 days p.i., at a time of the heterogeneous population in PBMCs. The viral population in VWCs was found to be a single clone and this clone was a minor type in PBMCs, suggesting that the major clone in PBMCs was not always secreted to the vaginal cavity. PMID- 9858468 TI - TbRab2p, a marker for the endoplasmic reticulum of Trypanosoma brucei, localises to the ERGIC in mammalian cells. AB - The Rab family of small GTPases is a subset of the Ras superfamily. Rabs regulate the flux through individual steps of the intracellular membrane trafficking pathway, such as ER-to-Golgi transport, probably by controlling SNARE complex assembly. In Trypanosoma brucei a number of Rab proteins have been isolated by EST analysis; here we characterise one of these, TbRab2p (originally designated Trab1p), which is a member of the Ypt1p subfamily of Rab proteins. Recombinant TbRab2p is capable of hydrolysing GTP and is post-translationally modified in vitro by addition of a geranylgeranyl prenyl group, properties of an authentic Rab GTPase. Antibodies against recombinant TbRab2p show that in trypanosomes TbRab2p is localised primarily to the endoplasmic reticulum (ER) and colocalises with BiP in wild-type trypanosomes. Over expression of TbRab2p in procyclic form T. brucei results in a cell population having a 40-fold increase in TbRab2p expression. In these cells biosynthesis of procyclin, a secretory pathway glycoprotein, is decreased, accompanied by an increase in general protein biosynthesis, suggesting that excess TbRab2p affects ER function. Heterologous expression of TbRab2p in COS cells resulted in targeting to the pre-Golgi transport intermediate (ERGIC), indicating that the targeting information is conserved between mammals and trypanosomes. Clustal and phylogenetic analyses support assignment of TbRab2p as a Rab2 homologue. In addition, over expression of TbRab2p in trypanosomes results in membrane reorganisation and formation of opaque vesicular structures visible by phase contrast microscopy, consistent with accumulation of ER-derived vesicular structures in cells highly overexpressing TbRab2p. Ultrastructural examination by electron microscopy confirmed the presence of a tubulo-vesicular membrane bound compartment in close proximity to the cis-Golgi, probably equivalent to the ERGIC. TbRab2p is therefore a new ER/ERGIC marker for T. brucei. PMID- 9858469 TI - Nuclear organisation of NIPP1, a regulatory subunit of protein phosphatase 1 that associates with pre-mRNA splicing factors. AB - Protein phosphatase-1 (PP1) is complexed to many proteins that target it to particular subcellular locations and regulate its activity. Here, we show that 'nuclear inhibitor of PP1' (NIPP1), a major nuclear PP1-binding protein, shows a speckled nucleoplasmic distribution where it is colocalised with pre-mRNA splicing factors. One of these factors (Sm) is also shown to be complexed to NIPP1 in nuclear extracts. Immunodepletion of NIPP1 from nuclear extracts, or addition of a 'dominant negative' mutant lacking a functional PP1 binding site, greatly reduces pre-mRNA splicing activity in vitro. These findings implicate the NIPP1-PP1 complex in the control of pre-mRNA splicing. PMID- 9858470 TI - Transdifferentiation of NRP-152 rat prostatic basal epithelial cells toward a luminal phenotype: regulation by glucocorticoid, insulin-like growth factor-I and transforming growth factor-beta. AB - The role of basal epithelial cells in prostatic function, development and carcinogenesis is unknown. The ability of basal prostatic epithelial cells to acquire a luminal phenotype was explored in vitro using the NRP-152 rat dorsal lateral prostate epithelial cell line as a model system. NRP-152, which was spontaneously immortalized and clonally derived, is an androgen-responsive and nontumorigenic cell line that has a basal cell phenotype under normal growth conditions. However, when placed in mitogen-deficient media, these cells undergo a dramatic morphological change to a luminal phenotype. Under these growth restrictive conditions, immunocytochemical analysis shows that NRP-152 cells acquire the luminal markers Z0-1 (a tight-junction associated protein), occludin (integral tight-junction protein), and cytokeratin 18, and lose the basal markers cytokeratins 5 and 14. Total protein and mRNA levels of cytokeratins 8, 18, c-CAM 105 (the calcium-independent cell adhesion molecule) and Z0-1, as detected by western and/or northern blot analyses, respectively, are induced, while cytokeratin 5 and 15 are lost, and occludin is unchanged. Concomitant with this differentiation, expression of transforming growth factor-beta2 (TGF-beta2), TGF beta3, and TGF-beta receptor type II (TbetaRII) is induced, while those of TGF beta1 and TbetaRI remain essentially unchanged. Mitogens, such as insulin-like growth factor-I and dexamethasone inhibit luminal differentiation, while exogenous TGF-beta induces such differentiation. These data together with TGF beta neutralization experiments using pan-specific antibody implicate an important role for autocrine TGF-beta in the induction of the luminal differentiation. PMID- 9858472 TI - The four populations of myoblasts involved in human limb muscle formation are present from the onset of primary myotube formation. AB - To understand how and when myogenic precursor cells become committed to their particular developmental programs, we have analysed the different populations of myoblasts which grow out from explants of muscle tissue isolated from human limb buds from the beginning of primary fibre formation throughout subsequent development and post-natal growth. Four phenotypically distinct types of myoblasts were identified on the basis of their expression of desmin, myogenin and myosin heavy chain isoforms (MyHC), and after 5 and 20 divisions, cells were cloned. All four types of myoblasts were present at the beginning of primary myogenesis. Each respective phenotype was stably heritable through cloning and subsequent proliferation. The type 1 clones correspond to a novel class of myoblasts never described during human development, that biochemically differentiates, but does not fuse. Type 2 clones are composed of small myotubes expressing only embryonic MyHC. Type 3 clones are composed of thin and long myotubes expressing both embryonic and fetal MyHCs. The type 4 clones are composed of myotubes that have a phenotype very similar to human satellite cells. Contrasting with others species, no other population of myoblasts appear during fetal development and only the relative number of these four types changes. PMID- 9858471 TI - Characterization of a focal adhesion protein, Hic-5, that shares extensive homology with paxillin. AB - Paxillin is a focal adhesion scaffolding protein which was originally identified as a substrate of the oncogenic tyrosine kinase, v-src. Paxillin has been proposed to be involved in regulation of focal adhesion dynamics. Two alternatively spliced mouse paxillin cDNAs were cloned and in the process, a paxillin-related protein, Hic-5, was also identified. Cloning and characterization of Hic-5 indicates that this protein shares extensive homology with paxillin. Although Hic-5 was originally characterized as a TGF-beta inducible gene and proposed to be a transcription factor involved in senescence, the studies here demonstrate that Hic-5 is localized to focal adhesion in REF52 cells and can interact with the focal adhesion proteins, Fak, Frnk, and vinculin. In addition, like paxillin, Hic-5 can bind to a negative regulator of Src PTKs, csk but does not bind to the adaptor protein Crk. Like paxillin, localization of this protein to focal adhesions is mediated primarily by the LIM domains; however, sequences outside the LIM domains also play a minor role in focal adhesion targeting. These results suggest that Hic-5 like paxillin could be involved in regulation of focal adhesion dynamics and raise the possibility that Hic-5 and paxillin could have overlapping or opposing functions in the overall regulation of cell growth and differentiation. PMID- 9858473 TI - Simple fluctuation of Ca2+ elicits the complex circadian dynamics of cyclic AMP and cyclic GMP in Paramecium. AB - The circadian dynamics of cyclic adenosine 3',5'-monophosphate (cAMP) and cyclic guanosine 3',5'-monophosphate (cGMP) were simulated in Paramecium multimicronucleatum. The mathematical functions determined closely mimic the Ca2+ dependence of adenylate cyclase (AC) and guanylate cyclase (GC) activities as documented in P. tetraurelia. Patterns of cAMP concentration ([cAMP]), cGMP concentration ([cGMP ]), and the ratio [cGMP]/[cAMP] were calculated with respect to Ca2+ concentrations ([Ca2+]) fluctuating sinusoidally with a period of 24 hours at three different levels: low, medium, and high. The functions displayed varying patterns of [cAMP] characteristic for [Ca2+] fluctuating at each level, while patterns of [cGMP] and [cGMP]/[cAMP] almost paralleled [Ca2+] fluctuations. Similar patterns were observed for actual [cAMP] and [cGMP] measured during the light/dark cycle in P. multimicronucleatum, grown in axenic media additionally containing [Ca2+] at 25 (low), 100 (medium), or 400 (high) microM, respectively. The coincidence between simulated and measured fluctuations of [cAMP] and [cGMP] suggests that the circadian fluctuations of intracellular [Ca2+] primarily stimulate activities of AC and GC via their different degrees of Ca2+ dependence, which are ultimately responsible for the circadian spatiotemporal organization of various physiological functions in Paramecium. PMID- 9858474 TI - Dystrophic phenotype induced in vitro by antibody blockade of muscle alpha dystroglycan-laminin interaction. AB - alpha-dystroglycan is a glycoprotein expressed on the surface of skeletal muscle fibres and other cell types. In muscle, alpha-dystroglycan provides a link between the myofibre cytoskeleton through its indirect binding to dystrophin, and the basal lamina through its binding to laminin-2, a protein of the extracellular matrix. The disruption of this linkage between the myofibre cytoskeleton and the extracellular matrix is a common feature of Duchenne and other muscular dystrophies, though the pathogenic mechanisms leading to muscle wasting remain unknown. By treating primary mouse muscle cultures with a monoclonal antibody which blocks alpha-dystroglycan binding to laminin, we show here the induction of a dystrophic phenotype in vitro. The phenotype is inducible in differentiated cultures only, is characterised by reduced myotube size, myofibril disorganisation, loss of contractile activity, reduced spontaneous clustering of acetylcholine receptors and is reversed by addition of excess exogenous laminin 2. Thus, alpha-dystroglycan may be part of a signalling pathway for the maturation and maintenance of skeletal myofibres. Detailed knowledge of this signalling pathway may provide insights into the molecular pathology of the various inherited muscular dystrophies, and identify valuable pharmacological targets and new therapeutic strategies. PMID- 9858475 TI - Amino acid motifs required for isolated beta cytoplasmic domains to regulate 'in trans' beta1 integrin conformation and function in cell attachment. AB - The role of beta cytoplasmic domains in regulating beta1 integrin conformation and function in cell attachment is not fully understood. In this study, we tested the ability of transiently expressed beta cytoplasmic domains connected to an extracellular reporter domain to regulate 'in trans' the conformation of endogenous beta1 integrins, and compared these effects on cell attachment. We found that chimeric receptors containing either the beta1, beta3 or beta5 cytoplasmic domains inhibited the expression of the conformationally dependent 9EG7 and 12G10 epitopes on endogenous beta1 integrins. In contrast, chimeric receptors containing the beta4 or alpha5 cytoplasmic domain, or a control receptor lacking a cytoplasmic domain, had no effect. This inhibition occurred in a dose-dependent manner that required high levels of expression of the chimeric receptor. These results suggest that beta1 integrin conformation can be regulated by conserved cytosolic interactions involving beta cytoplasmic domains. This is further supported by our findings that mutations within amino acid motifs conserved among these beta cytoplasmic domains, specifically the NXXY, NPXY and TST-like motifs, reduced the ability of these chimeric receptors to regulate beta1 integrin conformation. Interestingly, the chimeric receptors inhibited cell attachment in a similar dose-dependent manner and required intact NXXY, NPXY, and TST-like motifs. The beta1 chimera also inhibited the binding of soluble fibronectin to endogenous beta1 integrins. Thus, the concomitant inhibition in the expression of conformation-dependent integrin epitopes, cell attachment and ligand binding by the chimeras, suggests that the expression of the 9EG7 and 12G10 epitopes correlates with integrin function. However, Mn2+, which is an extracellular activator of integrin function, increased 9EG7 expression to basal levels in the presence of the beta1 chimera, but did not rescue cell attachment to the same extent. Thus, although the beta1 integrin conformation recognized by mAb 9EG7 may be required for cell attachment, it is not sufficient, suggesting that the beta chimeras may be inhibiting both ligand binding and post-ligand binding events required for cell attachment. In addition, the inhibitory effects of the chimeric receptors on cell attachment were not reversed by the addition of the pharmacological agents that inhibit intracellular signals previously shown to inhibit integrin function. This finding, together with the requirement for high levels of the chimeric receptors and the fact that mutations in the same conserved motifs in heterodimeric beta1 integrins have been reported to regulate beta1 integrin conformation and function in cell attachment, suggest that beta cytoplasmic domains regulate these processes by interacting with cytosolic factors and that the regulatory effect of the chimeras may be due to their ability to titrate proteins from endogenous integrins. PMID- 9858476 TI - Cytoskeletal changes induced by GRAF, the GTPase regulator associated with focal adhesion kinase, are mediated by Rho. AB - Graf, the GTPase regulator associated with focal adhesion kinase was previously shown to have GAP activity for &Rgr; A and Cdc42 in vitro (Hildebrand et al 1996 Mol. Cell Biol. 16: 3169-3178). In this study we sought to determine whether Graf acted at the level of Cdc42, Rho, or both in vivo and whether Graf was a signal terminator or transducer for these proteins. Microinjection of Graf cDNA into subconfluent Swiss 3T3 cells (in the presence of serum) has marked effects on cell shape and actin localization. Graf expression causes clearing of stress fibers followed by formation of long actin based filopodial-like extensions. Similar phenotypes were observed following injection of the Rho-inhibitor, C3 into these cells. The Graf response was dependent on GAP activity, since injection of Graf cDNA containing point mutations in the GAP domain (R236Q or N351V) which block enzymatic activity, does not confer this phenotype. Injection of Graf into Swiss 3T3 cells in which Rho has been down-regulated by serum starvation has no effect on cell morphology. Using this system, we demonstrate that Graf blocks sphingosine-1-phosphate (SPP) stimulated (Rho-mediated) stress fiber formation. Conversely, Graf expression does not inhibit bradykinin stimulated (Cdc42-mediated) filopodial extensions. These data indicate that Graf is a GAP for Rho in vivo. To further substantiate these results we examined the effect of Graf over-expression on Rho-mediated neurite retraction in nerve growth factor (NGF)-differentiated PC12 cells. In PC12 cells, which express relatively high levels of endogenous Graf, overexpression of Graf (but not Graf containing the R236Q mutation) enhances SPP-induced neurite retraction. These data indicate the possibility that Graf may be an effector for Rho in certain cell types. PMID- 9858477 TI - Role of collagenase in mediating in vitro alveolar epithelial wound repair. AB - Type II pneumocytes are essential for repair of the injured alveolar epithelium. The effect of two MMP collagenases, MMP-1 and MMP-13 on alveolar epithelial repair was studied in vitro. The A549 alveolar epithelial cell line and primary rat alveolar epithelial cell cultures were used. Cell adhesion and cell migration were measured with and without exogenous MMP-1. Wound healing of a cell monolayer of rat alveolar epithelial cell after a mechanical injury was evaluated by time lapse video analysis. Cell adhesion on type I collagen, as well as cytoskeleton stiffness, was decreased in the presence of exogenous collagenases. A similar decrease was observed when cell adhesion was tested on collagen that was first incubated with MMP-1 (versus control on intact collagen). Cell migration on type I collagen was promoted by collagenases. Wound healing of an alveolar epithelial cell monolayer was enhanced in the presence of exogenous collagenases. Our results suggest that collagenases could modulate the repair process by decreasing cell adhesion and cell stiffness, and by increasing cell migration on type I collagen. Collagen degradation could modify cell adhesion sites and collagen degradation peptides could induce alveolar type II pneumocyte migration. New insights regarding alveolar epithelial cell migration are particularly relevant to investigate early events during alveolar epithelial repair following lung injury. PMID- 9858478 TI - Induction of antisense Pax-3 expression leads to the rapid morphological differentiation of neuronal cells and an altered response to the mitogenic growth factor bFGF. AB - Mutations within the Pax-3 gene lead to a range of developmental abnormalities in both humans and mice. In this report, we have investigated the role that Pax-3 plays in neuronal cell development by specifically downregulating Pax-3 expression within a neuronal cell line. This was achieved by stably transfecting the neuronal cell line ND7 with an expression vector in which antisense Pax-3 RNA was produced under the control of the inducible MMTV promoter. In the stable transfectants, we found that the addition of dexamethasone led to the induction of antisense Pax-3 RNA and a rapid downregulation in endogenous Pax-3 protein expression. The decrease in endogenous Pax-3 protein expression corresponded with a dramatic change in the morphology of the cell: the normally rounded ND7 cells exhibited increased cell to substrate adhesion, extended long neurite processes and expressed genes such as snap-25 that are characteristic of a mature neuron. The morphological differentiation induced by a reduction in Pax-3 expression was followed 24-48 hours later by a cessation in cell proliferation. Interestingly the morphological differentiation and cessation in cell proliferation inducted in the cell lines lacking Pax-3 could be reversed by the addition of the mitogenic growth factor EGF but not by bFGF, whose receptor was downregulated in these cells. These results suggest that the expression of Pax-3 is essential to maintain the undifferentiated phenotype of these immature neuronal cells, and in its absence the cells acquire many of the characteristics of a mature neuronal cell. The slow onset of cell cycle arrest in the cells lacking Pax-3 argues against this transcription factor playing a direct role in the regulation of neuronal cell proliferation. PMID- 9858479 TI - Production of gelatinases and tissue inhibitors of matrix metalloproteinases by equine ovarian stromal cells In vitro. AB - Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) play very important roles in extracellular matrix (ECM) remodeling in ovarian follicle growth and ovulation. Equine follicles are embedded in cortex that is at the center of the ovary, and they must expand/emigrate to the fossa, the only site in the ovary for ovulation. Therefore, equine ovarian stromal cells (EOSC) are probably involved in ECM remodeling during follicle growth. This study examined whether cultured EOSC synthesize gelatinases and TIMPs, molecules essential for ECM remodeling in other systems. Results showed that cultured EOSC (passage 3-8) had a fibroblast-like morphology and were positive for alpha-smooth muscle actin and type I procollagen by immunostaining. Gelatinase A (MMP-2), gelatinase B (MMP 9), TIMP-1, and TIMP-2 were present in EOSC-conditioned medium, and TIMP-3 in ECM of EOSC. Transforming growth factor beta significantly stimulated the activity of gelatinases A and B and TIMP-1 in conditioned medium from EOSC (p < 0.05). Phorbol 12-myristate 13-acetate also significantly stimulated the activity of gelatinases A and B and TIMP-1 in conditioned medium and of TIMP-3 in ECM (p < 0.05). Our results suggest that EOSC produce important components of the ECM remodeling machinery and, therefore, may play a role in the ECM remodeling during follicle growth in this species. PMID- 9858480 TI - Identification of naturally occurring follistatin complexes in human biological fluids. AB - Follistatin (FS) binds activin and inhibin proteins. Many organs are sensitive to activin and inhibin; thus the formation of FS-activin/inhibin complexes is important to our understanding of ligand activity. Other investigators studying FS have detected large molecular weight immunoreactive FS bands (greater than the expected molecular weight of FS alone) that have not been well characterized. The goal of this study was to identify naturally occurring FS monomers and FS activin/inhibin complexes in several organ systems. The pituitary, ovary, kidney, and urine were chosen for this investigation. Molecular masses were assigned to in vitro assemblies of complexes containing recombinant inhibin or activin with FS for comparison with naturally occurring FS forms. The recombinant complex of FS-activin was primarily 97-kDa size, while FS-inhibin complexes were detected in a range of molecular sizes from 66 kDa to 97 kDa, 133 kDa, and > 220 kDa. FS containing complexes of 66-kDa, 97-kDa, and 133-kDa were identified in the tissues examined and in pregnant urine. Our study points to the assembly of a series of FS-activin/inhibin complexes in a variety of organ systems that may impact upon the available amount of free versus bound (or "complexed") ligand, which must be considered when investigating the biology of activin- or inhibin responsive cells. In addition, urine may be an important biological fluid that can be used to measure significant changes in circulating FS complexes. PMID- 9858481 TI - Stage-dependent accumulation of cadmium and induction of metallothionein-like binding activity in the testis of the Dogfish shark, Squalus acanthias. AB - Cadmium (Cd) is an established spermatotoxicant. Using the shark (Squalus acanthias) testis model, we investigated stage-related patterns of intratesticular Cd accumulation and effect. After a single injection of 109CdCl2, tracer was rapidly eliminated from plasma but accumulated and was retained in testis for at least 7 days. Intratesticular 109Cd was stage dependent, resulting in a 3- to 5-fold gradient: germinal zone (GZ) > premeiotic (PrM) > meiotic (M) > postmeiotic (PoM) stages. When measured as tissue:plasma ratios, the Cd-binding mechanism in GZ (71:1) was similar to that in liver (87:1) but lower than in kidney (381:1). The same intratesticular gradient was seen in untreated controls when tissue Cd levels were measured by atomic absorption spectroscopy, implying environmental exposure. A single CdCl2 injection (5 mg/kg i.v.) elevated testicular Cd > 160-fold in all stages but did not alter the direction or magnitude of the gradient. Intratesticular distribution of metallothionein-like Cd-binding protein was stage dependent (PrM = PoM > GZ = M), but the pattern differed from the Cd gradient. This binding component was Cd inducible in all but M stages, but induction did not alter the stage-dependent pattern of binding activity or Cd accumulation. Analysis of tissue subfractions after in vivo tracer injection indicated that the binding mechanism responsible for the intratesticular gradient is mainly cytosolic, but that a second less abundant component is associated with the nucleus. The functional significance of preferential Cd accumulation in GZ and PrM stages of spermatogenesis remains to be determined. PMID- 9858483 TI - Sperm subpopulations in boar (Sus scrofa) and gazelle (Gazella dama mhorr) semen as revealed by pattern analysis of computer-assisted motility assessments. AB - The aim of this study was to test the suitability of "pattern analysis" for the exploration of data provided by computer-assisted semen analysis methods. Data sets derived from the examination of boar sperm responses to bicarbonate and caffeine (measurements on 3208 spermatozoa) and from studies of semen cryopreservation in Mohor gazelles (7278 spermatozoa) were reanalyzed. A nonhierarchical classification method was used to generate initial subgroups of spermatozoa (9 for boar, 13 for gazelle). The subgroup centroids were fused, yielding three boar sperm subpopulations and four gazelle sperm subpopulations distinguished by sperm behaviors. Bicarbonate and caffeine both induced major transitions (p < 0.0001) of boar sperm behavior, detected as shifts in group membership (from group 2, i.e., active but nonlinear movement, into group 1, i.e., linear, rapid movement). Some spermatozoa (approximately 3%) were refractory to both caffeine and bicarbonate. The gazelle sperm subpopulation structure was affected by the inclusion of equex (sodium triethanolamine lauryl sulfate) in the cryoprotective diluents. Equex suppressed the appearance of spermatozoa with erratic behavior (p < 0.0001; high curvilinear velocity, low linearity, low straight-line velocity) after cryopreservation. The proportion of these erratic spermatozoa was positively correlated with animal age (r = 0.68, p = 0.029). Pattern analysis revealed novel aspects of the data not seen in the original investigations and usefully supplemented the more standard data analysis approaches. PMID- 9858482 TI - Repression of major histocompatibility complex genes by a human trophoblast ribonucleic acid. AB - The suppression of polymorphic major histocompatibility complex antigen expression in human trophoblasts is critical for the avoidance of a cell-mediated immune response by maternal lymphocytes against cells expressing paternal antigens. In this study, a repressor of major histocompatibility complex gene expression was cloned by negative immunoselection using a trophoblast cDNA expression library in interferon-gamma-responsive human cells. The sequence of this regulatory gene was analyzed, and the functions of the transfected cDNA or microinjected gene product were examined in interferon-gamma-responsive cells by immunocytochemical methods. The repressor, called TSU-- trophoblast STAT (signal transducers and activators of transcription) utron (untranslated region of an mRNA)--reduced STAT1 nuclear translocation and suppressed major histocompatibility complex class II antigen expression at high doses of interferon-gamma and class I expression at low doses of interferon-gamma. TSU encoded a small, untranslated poly-A+-RNA that appeared to bind STAT1 through pairs of motifs analogous to STAT-binding promoter sequences. These promoter-like motifs, but no open reading frame, were conserved in a TSU-related gene in goats. Northern blot analysis demonstrated that TSU was expressed as a 0. 5-kilobase (kb) RNA in placenta and as an ubiquitous 4.4-kb RNA. TSU expression may protect trophoblasts from immune attack and promote the survival of the placenta and fetus. PMID- 9858484 TI - Effects of porcine follicular fluid and oviduct-conditioned media on maturation and fertilization of porcine oocytes in vitro. AB - Advances in porcine in vitro fertilization have been impaired by low normal fertilization rates resulting from a high rate of polyspermy. The present study was undertaken to determine the effects of porcine follicular fluid (pFF) and oviductal explant-conditioned medium on maturation and fertilization of porcine oocytes in vitro. Oocytes and pFF were collected from small, medium, and large follicles and pooled within size category. Maturation and fertilization media were supplemented (10%) with either fetal calf serum (FCS) or pFF (either fresh or snap-frozen). Snap-frozen pFF from small (3.1-5.0 mm) and medium (5.1-7 mm) follicles, respectively, increased maturation rates of oocytes from small and medium follicles by nearly 36% (p < 0.05) compared with those treated with FCS or fresh pFF. Supplementing media with either fresh or snap-frozen pFF from medium follicles reduced (p < 0.05) polyspermy of oocytes from small follicles by 30% compared with supplemental FCS. Snap-frozen pFF increased (p < 0.05) normal fertilization compared to that in fresh pFF (29% vs. 18%). Supplementing oocytes from medium follicles with snap-frozen pFF yielded the lowest (18%, p < 0.05) polyspermy rate. Oocytes from both small and medium follicles supplemented with pFF and/or conditioned medium (CM) from oviducts of periovulatory gilts exhibited a 95% improvement in normal fertilization rate and a 34% decrease in polyspermy rate compared to those treated with FCS (p < 0.05). CM from oviducts of luteal gilts did not improve rates of polyspermy and normal fertilization (p > 0.05). We conclude that snap-frozen follicular fluid from medium follicles and CM from cultured oviducts of periovulatory gilts improve in vitro maturation, reduce polyspermy, and increase normal fertilization rates in vitro. PMID- 9858485 TI - Differential distribution of inositol trisphosphate receptor isoforms in mouse oocytes. AB - In mammalian fertilization, inositol 1,4,5-trisphosphate receptor (IP3R) dependent Ca2+ release is a crucial signaling event that originates from the vicinity of sperm-egg interaction and spreads as a wave throughout the egg cytoplasm. While it is known that Ca2+ is released by the type 1 IP3R in the egg cortex, the potential involvement of other isoform types responsible for the Ca2+ rise in the mouse egg (interior) and their spatial distribution are not known. In addition, the biochemical basis has not been definitively established for the development of increased sensitivity to inositol 1,4,5-trisphosphate (IP3) during meiotic maturation. Using specific antibodies to the type 1, 2, and 3 IP3R, we tested the hypotheses that different IP3R isoforms are responsible for the internal Ca2+ elevation and that they contribute to the maturation-associated acquisition of IP3 sensitivity. In both preovulatory oocytes and ovulated eggs of CF-1 mice, immunofluorescence revealed that types 1 and 2 isoforms were present in the cell cortex and interior. Type 1 was observed throughout the cytoplasm, and Western analysis indicated a 1.9-fold maturation-associated increase. In contrast, the signals detected for the type 2 (high-affinity) isoform and type 3 were present to a lesser extent, with type 2 restricted to isolated islands (similar to aggregates of vesicles detected by electron microscopy), which, in the cortex, may amplify early sperm-egg signaling events. The cortical-to perinuclear localization of the receptor and cortical vesicle aggregates imply an efficient mechanism for propagating Ca2+ release from the cortex into the interior of the egg to activate development, and the isoform localization analysis indicates a clear spatial and biochemical heterogeneity. Types 1 and 2 isoforms were also present in granulosa cells. PMID- 9858486 TI - MUC4 and MUC5B transcripts are the prevalent mucin messenger ribonucleic acids of the human endocervix. AB - Mucins secreted by the endocervical epithelium protect the surfaces of the reproductive tract epithelium from pathogen penetrance and modulate sperm entry into the uterus. Three large gel-forming mucins, MUCs 5AC, 5B, and 6, are expressed by the endocervical epithelium, as is MUC4, a relatively uncharacterized mucin for which only tandem repeat sequence has been reported. We sought to determine the relative abundance of each of these mucin gene transcripts and to relate their expression to blood progesterone and estradiol. Samples were obtained from six subjects at successive stages in the menstrual cycle. Primers to nontandem repeat sequences of MUCs 4, 5AC, 5B, and 6 were used in semiquantitative reverse transcription-polymerase chain reaction to determine relative abundance of each mucin gene in relation to beta2-microglobulin message control. In order to design primers from a nontandem repeat region of MUC4 so that MUC4 message levels could be quantitated, we obtained approximately 2.7 kilobase nontandem repeat sequence 5' to the tandem repeat sequence of a MUC4 genomic clone. The sequence showed lack of cysteine-rich D-domains and was rich in serine and threonine. Semiquantitative polymerase chain reaction analyses indicated that the principal mucin transcripts of human endocervix are MUC4 and MUC5B, with MUC4 predominant in 15 of 21 samples. When correlated with plasma steroid levels, message levels of both MUC4 and MUC5B were inversely related to progesterone levels. PMID- 9858487 TI - Concurrent pulsatile secretion of luteinizing hormone and follicle-stimulating hormone during different phases of the estrous cycle and anestrus in beagle bitches. AB - The 6-h secretory profiles of LH and FSH and the possible concordance between the episodic release of LH and FSH were studied in 6 beagle bitches during early, mid , and late anestrus and during the follicular and luteal phases of the estrous cycle. Plasma samples were obtained at 10-min intervals via jugular venipuncture. In all stages of anestrus and in the luteal phase, FSH and LH secretion was pulsatile. All FSH pulses coincided with LH pulses. However, the mean duration of the FSH pulse (115 min) was significantly longer than that of the LH pulse (72 min). The basal plasma LH concentration was low compared with the maximum peak levels, whereas FSH pulses were characterized by relatively low peaks compared with the basal levels. In contrast to the basal plasma LH levels and the area under the curve (AUC) for LH, the basal plasma FSH levels and the AUC for FSH increased significantly as anestrus progressed. During the follicular phase, the secretory pattern of LH was characterized by frequent increases of short duration. During this phase, the basal plasma FSH concentration was relatively low, whereas the basal plasma LH level was high in comparison with that in the other phases of the estrous cycle. The luteal phase was characterized by an increased frequency of LH pulses, a shorter duration of the LH peaks, and a tendency to a lower amplitude of both LH and FSH peaks compared with values observed during anestrus. It is concluded that in the bitch, FSH and LH pulses are released in concordance and that progression from early to late anestrus is associated with an increase in basal plasma FSH concentration without a concomitant rise in basal plasma LH concentrations. The latter suggests that in the bitch an increase in circulating FSH should be considered to be a critical event required for the initiation of ovarian folliculogenesis and consequently for the termination of anestrus. PMID- 9858488 TI - Relationship between follicular development and the decline in the follicle stimulating hormone surge in heifers. AB - Experiment 1 was conducted to determine whether progesterone affects the pattern of the FSH surge or follicular development associated with a follicular wave in heifers. On Day 7 (Day 0 = ovulation), heifers were allocated into a group receiving prostaglandin F2alpha (PGF2alpha; n = 6) or a control group (n = 5). Twenty-four hours later, all detectable follicles (>/= 2 mm) were ablated (Hour 0). Follicular development was monitored Hours 0, 3, 6, 9, 12, and 16, at 8-h intervals thereafter until Hour 112. To monitor FSH concentrations, blood was sampled at Hours -24, 0, 3, 6, 9, 12, and 16, and at 8-h intervals thereafter until Hour 104. There were no differences (p > 0.05) between the PGF2alpha treated group and controls in the patterns of the FSH surge or follicular development. Experiment 2 tested the hypothesis that 3-mm follicles do not have FSH-suppressing capacity and that suppression increases as follicles grow beyond 5 mm. Twenty-four hours after an injection of PGF2alpha (Days 6-8), heifers were subjected to either ablation of follicles >/= 2 mm or ovariectomy. Intact heifers were allocated into four groups (n = 5) in which all follicles of the new wave were ablated upon reaching either 3, 5, or 7 mm or were not ablated (controls). Blood was sampled at 8-h intervals to monitor FSH and estradiol-17beta. Averaged over Hours 8-120, FSH concentrations (ng/ml) were higher (p < 0.05) in the ovariectomized (2.02 +/- 0.05) and the 3-mm groups (1.91 +/- 0.05) than in the 5 mm (1.52 +/- 0.05), 7-mm (1.35 +/- 0.04), and control groups (1.33 +/- 0.05); and estradiol concentrations (pg/ml) were lower (p < 0.05) in the ovariectomized group (0.19 +/- 0.03) than in the 3-mm (1.48 +/- 0. 16), 5-mm (1.56 +/- 0.15), 7 mm (2.22 +/- 0.27), and control groups (2.55 +/- 0.49). In conclusion, the presence of endogenous progesterone did not affect FSH patterns or follicular development. Follicles pimozide (8 microM) > mibefradil (11 microM). By contrast, the effects of these drugs on the P4-initiated AR were varied: pimozide (10 and 20 microM) significantly (p < 0.05) increased the percentage of AR spermatozoa, calmidazolium was without effect, and mibefradil (20 microM) significantly (p < 0.05) inhibited the AR. These disparate results do not allow us to reach any definitive conclusion concerning the role of a sperm VOCCT in the mechanism of the P4-initiated AR. However, the differences between the [Ca2+]i and AR effects, in particular the inverse relationship in the case of pimozide, suggest a dissociation between the amplitude of the P4-stimulated Ca2+ signal and the downstream biological effect of that signal, the AR. PMID- 9858493 TI - Synthesis of retinoic acid by rat ovarian cells that express cellular retinoic acid-binding protein-II. AB - The induction of pseudopregnancy by the injection of eCG in rats results in the appearance of cellular retinoic acid-binding protein type II (CRABP[II]) in the granulosa cells of the ovary and the lining epithelium of the uterus within 48 h. This expression pattern is also seen in the normal mature female rat, in which CRABP(II) is expressed in the uterine epithelium during estrus (but not diestrus) and in the granulosa and luteal cells of the ovary. We have previously demonstrated that the uterine epithelial cells from the pseudopregnant rat have gained the ability to synthesize retinoic acid from retinol, in correlation with the induced expression of CRABP(II). If this is true for other sites of CRABP(II) expression, then local production of retinoic acid is intimately connected with various stages of reproduction in the female. Here we report that granulosa cells from the ovary of the eCG-treated immature rat and luteal cells from the ovary of the eCG/hCG-treated immature rat (both of which express CRABP[II]) synthesized markedly higher amounts of retinoic acid when cultured, compared to granulosa cells cultured from the ovary of the prepubertal rat treated with control vehicle. Culturing the granulosa cells from either control or eCG-treated animals had no effect on the expression of CRABP(II) cells. These data are consistent with our hypothesis that CRABP(II) expression is associated with retinoic acid synthesis and strengthen the case that local generation of retinoic acid plays an important role in reproduction. PMID- 9858494 TI - Testicular morphology and function in boars differing in concentrations of plasma follicle-stimulating hormone. AB - The objective of this study was to evaluate morphological characteristics and testicular function of boars with different endogenous concentrations of FSH. Boars were selected at 6 mo of age on the basis of mean FSH concentrations in plasma collected at 4, 5, and 6 mo of age. Boars were classified within half sibling families based on whether they had high concentrations of FSH (HiFSH, > 500 ng/ml, n = 9) or low concentrations (LoFSH, < 500 ng/ml, n = 7). At 14.5 mo, testes were collected, fixed, sectioned at 1 microm, and evaluated for morphological characteristics. Boars with LoFSH had larger (p < 0.01) testicular and epididymal weights than boars with HiFSH, greater (p < 0.01) daily sperm production per gram of testis, and greater total daily sperm production per boar. Testes of boars with LoFSH had a greater (p < 0.03) volume percentage of seminiferous tubules, a lesser percentage (p < 0.03) of Leydig cells, and a somewhat lesser (p = 0.06) percentage of vascular structures than testes of boars with HiFSH. Testes of boars with LoFSH had greater (p < 0.01) total tubule volume and tubule length than testes of boars with HiFSH. There were no differences (p > 0.70) in volume, diameter, or total number of Leydig cells or in total interstitial volume in testes (p > 0.41) of these two groups. Production of testosterone in vitro per paired testis and per million Leydig cells was not different (p > 0.65) between boars with HiFSH or LoFSH. Greater concentrations of FSH in blood plasma were negatively associated with development of seminiferous tubules and spermatogenic efficiency, whereas Leydig cell development was not different in boars of these two groups. PMID- 9858495 TI - Development and differentiation of the interstitial and tubular compartments of fetal porcine testes. AB - Male differentiation is initiated by fetal testicular androgen synthesis, catalyzed by the enzyme 17alpha-hydroxylase/17,20 lyase cytochrome P450 (P450c17). This study was an investigation of testicular development and differentiation in porcine fetuses recovered on Days 30-42 of gestation. The expression of P450c17 was localized in fetal gonads by in situ hybridization and immunocytochemistry and related to cellular proliferation through expression of the proliferating cell nuclear antigen (PCNA). Gonadal P450c17 expression was quantified by Western immunoblot analysis and related to testosterone secretion by cultured explants of fetal gonads. P450c17 transcripts were detected in the interstitium surrounding testicular cords preceding the appearance of the enzyme protein. The intensity of both P450c17 hybridization and staining was greater in Yorkshire fetal gonads, which also exhibited more advanced tubular development. PCNA staining was prominent within tubular primordia and was higher in testes from Yorkshire than from Meishan fetuses on all days examined. P450c17 expression paralleled testosterone secretion, which decreased by Day 42, and was generally less in cultures of Meishan than of Yorkshire fetal gonads. These data demonstrate that the expression of P450c17 in porcine fetal testes coincides with differentiation of central medullary cells and androgen secretion during gonadal development between Days 30 and 42 of gestation. This occurs as medullary cords organize and is associated with changes in cellular proliferation within the tubular compartment. PMID- 9858496 TI - Seasonal variation in long-day stimulation of prolactin secretion in ewes. AB - Whereas ewes initiate reproductive activity in response to a photoperiod signal initiated after the winter solstice of 35 long days (35 LD) followed by short days, the reproductive axis fails to respond to this signal between the autumn equinox and the winter solstice. The aim of experiment 1 was to determine whether the prolactin axis, like the reproductive axis, is unresponsive to a 35 LD photoperiod signal followed by continuous exposure to short days between the autumn equinox and the winter solstice. Whereas the 35 LD signal from September 21 ( 6 h increase in day length) initiated a rise in prolactin in at least 75% of ewes in each group (p < 0.05). The aim of experiment 2 was to determine whether ewes failed to secrete prolactin during a 35 LD photoperiod from September 21 because they did not recognize a 6-h increase in day length at any time of year as a stimulatory photoperiod signal or because hypothalamic/pituitary regulation of prolactin synthesis or secretion is compromised in September. The results demonstrated that while hypothalamic regulation of prolactin secretion and pituitary stores of prolactin were normal at all times of year examined, the ability of ewes to secrete prolactin in response to a long-day photoperiod signal appears to be dependent on photoperiodic history rather than the time of year of the photoperiodic challenge. PMID- 9858497 TI - Estradiol up-regulates estrogen receptor-alpha messenger ribonucleic acid in sheep endometrium by increasing its stability. AB - During the preovulatory period, estrogen up-regulates estrogen receptor-alpha (ER) gene expression in endometrium in female mammals of all species examined. The purpose of this study was to determine directly whether estradiol up regulates ER mRNA by increasing the stability of the message. Endometrial tissue was collected from ovariectomized ewes 18 h after the ewes were injected with 50 microg estradiol. Previous work indicated rapid accumulation of ER mRNA at this time. Estradiol increased uterine weights (to 157 +/- 15%) as well as steady state concentrations of ER (to 309 +/- 37%), progesterone receptor (PR; to 165 +/ 19%), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; to 374 +/- 32%) mRNAs in endometrium, compared to control levels of 100%. The effects of estradiol on ER mRNA stability in endometrium were measured in explants cultured with the transcription inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole, as well as by labeling RNA in vivo with 4-thiouridine. Both assays indicated that estradiol enhanced ER mRNA stability (half-life increased from 9 h to >/= 24 h). The estradiol effect was specific, because the stabilities of PR, GAPDH, and c fos mRNAs were unaffected by treatment. Thus, estradiol up-regulates steady-state concentrations of ER mRNA in endometrium by a novel posttranscriptional mechanism. PMID- 9858498 TI - Yolk formation and degradation during oocyte maturation in seabream Sparus aurata: involvement of two lysosomal proteinases. AB - Oocyte growth within the follicle is preponderantly due to the accumulation of hepatically derived yolk protein (vitellogenin, VTG) by receptor-mediated endocytosis; once in the oocyte, VTG is partially processed and stored in yolk globules. In some pelagic egg-laying marine teleosts, additional cleavages of yolk proteins followed by a pronounced water uptake occur concomitantly with final oocyte maturation. The aim of this study was to establish the lysosomal enzymes involved in these two proteolytic processes that characterize oocyte maturation of seabream Sparus aurata. The enzymatic activities of several cathepsins were assessed in the various classes of oocytes. Changes in cathepsin B, D, and L activity were found depending on the oocyte maturation stage; cathepsin B and D were found to be at maximum level in early-vitellogenesis oocytes, and cathepsin L in mid-vitellogenesis ones. Cathepsin D and L were purified from seabream ovary, and their roles in VTG and lipovitellin (LV) proteolysis, respectively, were analyzed. Here we demonstrate directly that one of the catalysts for the intraoocytic processing of VTG in yolk proteins is cathepsin D; however, we cannot exclude also a role of cathepsin B in the same process. On the other hand, cathepsin L is responsible for the second proteolytic cleavage of the LV components. We postulate that the acquisition of buoyancy by eggs through the hydration process may be regulated by enzymatic activation at the appropriate time of oocyte maturation, this process probably being the key event in the reproduction of this marine pelagic egg spawner. PMID- 9858499 TI - Hormonal control of H-type alpha(1-2)fucosyltransferase messenger ribonucleic acid in the mouse uterus. AB - The H epitope, an alpha(1-2)fucosylated carbohydrate structure, has been implicated in initial attachment of the murine blastocyst to luminal uterine epithelial cells in vitro. In this study, the expression of the H-type alpha(1 2)fucosyltransferase (FUT1) gene was examined in endometrium of mice. Northern blotting of luminal epithelial RNA identified a single 6.2-kilobase transcript. In situ hybridization studies showed a signal for FUT1 mRNA on Days 1-3 of pregnancy in glands and luminal epithelium. The signal diminished by Day 4 and could not be detected on Day 5 of pregnancy. The in situ signal in endometrial epithelia was highest at estrus and metestrus and was absent at diestrus. Estrogen treatment after ovariectomy gave strong FUT1 mRNA expression in epithelia, but with progesterone, progesterone + estrogen, or vehicle, no message could be detected. A semiquantitative reverse transcription-polymerase chain reaction (PCR) analysis of FUT1 mRNA from luminal epithelium generated large amounts of PCR product on Day 1 of pregnancy; this diminished on Days 2, 3, and 4, and the product was barely detectable on Day 5. A kinetic analysis of FUT1 activity on Day 1 of pregnancy suggested a single enzyme with a Michaelis-Menten constant (Km) of 0.29 mM towards phenyl-beta-D-galactoside and of 1.75 mM towards Galbeta(1-3)GalNAc. These results suggest that expression of the H epitope is regulated at the level of FUT1 transcription and that transcription is stimulated by estrogen in the endometrial epithelium. PMID- 9858500 TI - Effect of stress-like concentrations of cortisol on gonadotroph function in orchidectomized sheep. AB - The effect of stress-like concentrations of cortisol (C) on the feedback potency of estradiol (E2) was assessed using 32 orchidectomized sheep (wethers) assigned at random to 1 of 4 treatment groups in a 2 x 2 factorial design. Wethers received C (3. 6 mg/50 kg per hour; groups 2 and 4) or a comparable volume of C delivery vehicle (groups 1 and 3) as a continuous infusion for 7 days. During the final 48 h of infusion, wethers received E2 (0.3 microg/50 kg/h; groups 3 and 4) or E2 delivery vehicle (groups 1 and 2). The pattern of LH secretion was assessed during a 4-h period of intensive blood collection beginning 44 h after initiation of E2 infusion. Gonadotroph responsiveness (LH secretion induced by GnRH challenge [500 ng, i.v.]) was determined 48 h after E2 delivery was begun. Although the frequency of secretory episodes of LH was not affected (p > 0.05) by infusion of C or E2 alone, LH pulse frequency was significantly decreased in wethers receiving C and E2 in combination. In contrast, neither the magnitude of basal gonadotroph responsiveness nor the extent of E2-dependent augmentation of responsiveness was significantly affected by stress-like concentrations of C. In a second experiment, the effect of C on the magnitude of E2-induced increase in pituitary concentration of GnRH receptor and GnRH receptor mRNA was assessed using 32 additional wethers. Continuous infusion of E2 for 48 h increased (p < 0.05) tissue concentrations of GnRH receptor and GnRH receptor mRNA. Concurrent delivery of C did not affect (p > 0.05) E2-induced increase in GnRH receptor mRNA but significantly reduced the magnitude of the E2-dependent increase in pituitary concentration of GnRH receptor. Collectively, these data indicate that stress like concentrations of C enhance the negative feedback potency of E2 and reduce estrogen-dependent augmentation of the concentration of GnRH receptor in pituitary tissue. PMID- 9858501 TI - Effect of stress-like concentrations of cortisol on estradiol-dependent expression of gonadotropin-releasing hormone receptor in orchidectomized sheep. AB - The effect of stress-like concentrations of cortisol (C) on estrogen-dependent expression of GnRH receptor was evaluated using orchidectomized sheep (wethers; n = 6 animals per group). C (5.0 mg/50 kg per hour; groups 1-4) or a comparable volume of vehicle (groups 5-8) was delivered by continuous infusion for 48 h. During the final 24 h of infusion, animals received concurrent infusion of estradiol (E2) at rates of 0 (groups 1 and 5), 0.5 (groups 2 and 6), 2.0 (groups 3 and 7), or 8.0 (groups 4 and 8) microg/50 kg per hour. Pituitary tissue was collected at the end of infusion. Although C did not affect (p > 0.05) the basal concentration of GnRH receptor or GnRH receptor mRNA, it reduced (p < 0.05) the increase in receptor and receptor mRNA induced by concurrent administration of 0. 5 microg E2/50 kg per hour. In contrast, the increase in GnRH receptor expression induced by higher levels of estrogen stimulation was not affected (p > 0.05) by concurrent administration of C. The effect of C on the temporal pattern of E2 dependent increase in GnRH receptor expression was assessed using wethers receiving E2 (0.5 microg/50 kg per hour) by continuous infusion for 0 (groups 1 and 5), 24 (groups 2 and 6), 48 (groups 3 and 7), or 72 h (groups 4 and 8). Animals received C (5.0 mg/50 kg per hour; groups 1-4) or vehicle (groups 5-8) beginning 24 h before, and continuing throughout, the E2 delivery period. Stress like concentrations of C reduced (p < 0. 05) the increase in GnRH receptor and receptor mRNA induced after 24 h of E2 stimulation. However, the suppressive effect of C was transient, and tissue levels of GnRH receptor and receptor mRNA were comparable after 72 h of E2 infusion in animals receiving C or vehicle alone. Collectively these observations demonstrate that C suppresses estrogen dependent increase in tissue concentrations of GnRH receptor and receptor mRNA. However, this effect of C is transient and not evident in animals receiving moderate to high levels of estrogen stimulation. This transient suppression of GnRH receptor expression may account, at least in part, for the anti-gonadal effect of glucocorticoids. PMID- 9858502 TI - Heparin and high-density lipoprotein mediate bovine sperm capacitation by different mechanisms. AB - Capacitation is an important process in bovine sperm maturation and is an obligatory step prior to fertilization. Two capacitating agents, namely heparin and high-density lipoprotein (HDL), have been shown to induce sperm capacitation. A family of major proteins of bovine seminal plasma designated BSP-A1/A2, BSP-A3, and BSP-30 kDa (collectively called BSP proteins) bind to the sperm surface upon ejaculation via their membrane choline phospholipids. Our previous studies with bovine epididymal sperm showed that BSP proteins potentiate sperm capacitation induced by heparin and HDL. This study was undertaken to clarify the mechanism of capacitation induced by heparin and HDL in the presence of BSP proteins. Washed bovine ejaculated sperm were incubated with heparin (12 microg/ml) or HDL (10-160 microg/ml) in the presence of polyclonal antibodies against purified BSP proteins (anti-BSP proteins). The percentage of capacitated sperm was evaluated after the induction of the acrosome reaction (AR) with lysophosphatidylcholine. When sperm were incubated for 5 h with heparin and anti-BSP proteins (40 microg/ml), the AR level was not significantly different from control levels (16. 8 +/- 0.9% vs. 12.9 +/- 0.9%). In contrast, incubation of sperm for 8 h with HDL and anti-BSP proteins did not inhibit the AR (42.4 +/- 1.1% vs. 17.1 +/- 1.6 for the control samples). We also investigated the effect of heparin and HDL on protein tyrosine phosphorylation associated with capacitation. The tyrosine phosphorylation of a group of proteins was increased in the presence of heparin. However, HDL did not significantly stimulate protein phosphorylation. The increase in phosphorylation was correlated with an increase in the AR after the incubation with heparin but not with HDL. These results indicate that heparin and HDL mediate capacitation via different mechanisms. PMID- 9858503 TI - Rat amnion type IV collagen composition and metabolism: implications for membrane breakdown. AB - We report here that rat amnion type IV collagens are composed primarily of alpha1(IV) and alpha2(IV) chains. Amnion basement membrane collagens were more sensitive to degradation by collagenases than were adult rat kidney basement membrane collagens, which are enriched in alpha3(IV), alpha4(IV), and alpha6(IV) chains. Amnion type IV collagen content per unit of protein was markedly reduced by Day 21 of pregnancy, the day of delivery. Increased amnion levels of matrix metalloproteinase (MMP)-2 and MMP-9, gelatinases that degrade type IV collagen, were found by Day 21, suggesting that collagen breakdown was responsible, in part, for the decline in amnion type IV collagen. Infection of organ cultures of Day 18 rat amnions with a recombinant adenovirus expressing MMP-9 (AdMMP-9) caused release of collagen fragments detected as hydroxyproline in the culture fluid, amnion cell detachment, and apoptosis. The AdMMP-9-induced apoptosis was prevented by the MMP inhibitor batimastat. These findings suggest that MMPs are implicated in anoikis and apoptotic death of amnion cells, and may be part of a complex program of fetal membrane remodeling that occurs before delivery. PMID- 9858504 TI - Induction of matrix metalloproteinases and collagenolysis in chick embryonic membranes before hatching. AB - The membranes surrounding the chick embryo undergo striking morphological changes before hatching, which include structural degradation of the allantoic membrane. The fibrillar collagen content of the membranes declined by embryonic day (ED) 20 (the day of hatching). By ED 19, a 55-kDa matrix metalloproteinase (MMP) activity appeared in the extraembryonic fluid, and by ED 20 there was substantial 55-kDa MMP activity in embryonic membrane extracts. Reverse transcription-polymerase chain reaction was employed to clone a partial cDNA representing the chicken homologue of MMP-13, a 55- to 57-kDa enzyme. MMP-13 mRNA dramatically increased in abundance in embryonic membranes by ED 19, reaching a peak on ED 20. Introduction of the MMP inhibitor batimastat into the extraembryonic fluid prevented the structural changes in the embryonic membranes before hatching. We conclude that, like mammalian fetal membranes, chick embryonic membranes undergo terminal remodeling before hatching, in part as a result of increased MMP activity. The chicken egg system represents a novel in vivo model for exploring biochemical events leading to embryonic membrane remodeling prior to birth and to test inhibitors of MMPs for their ability to prevent collagenolysis and fetal membrane rupture. PMID- 9858505 TI - Insulin-like growth factor-2 regulation of conceptus composition: effects of the trophectoderm and inner cell mass genotypes in the mouse. AB - The purpose of this study was to measure the effect of insulin-like growth factor 2 deficiency on the growth of the mouse conceptus. Initial observations on normal development in the 129J/Sv strain established that wet and dry weights were reduced by 35% when the insulin-like growth factor-2 gene was inactive. The DNA contents were reduced by only 15%. We exchanged the inner cell mass and trophectoderm between mouse blastocysts that had or lacked an active insulin-like growth factor-2 gene. At embryonic Day 16.5, lack of this gene's activity in the derivatives of either tissue decreased the fluid volumes of the exocelomic and amniotic cavities. The wet weights of the "fetal placentas," the yolk sacs, and the fetuses were also decreased. However, the tissue wet weight decrease could not be accounted for by the change in DNA content, indicating that cell associated biomass had changed. The conclusions are 1) that insulin-like growth factor-2 levels regulate the composition of the fetus and extra-embryonic tissues and 2) that trophectoderm and inner cell mass derivatives cooperate to control extra-cellular fluid volume in the conceptus. PMID- 9858506 TI - The critical need for CD4 help in maintaining effective cytotoxic T lymphocyte responses. PMID- 9858507 TI - Viral immune evasion due to persistence of activated T cells without effector function. AB - We examined the regulation of virus-specific CD8 T cell responses during chronic lymphocytic choriomeningitis virus (LCMV) infection of mice. Our study shows that within the same persistently infected host, different mechanisms can operate to silence antiviral T cell responses; CD8 T cells specific to one dominant viral epitope were deleted, whereas CD8 T cells responding to another dominant epitope persisted indefinitely. These virus-specific CD8 T cells expressed activation markers (CD69(hi), CD44(hi), CD62Llo) and proliferated in vivo but were unable to elaborate any antiviral effector functions. This unresponsive phenotype was more pronounced under conditions of CD4 T cell deficiency, highlighting the importance of CD8- CD4 T cell collaboration in controlling persistent infections. Importantly, in the presence of CD4 T cell help, adequate CD8 effector activity was maintained and the chronic viral infection eventually resolved. The persistence of activated virus-specific CD8 T cells without effector function reveals a novel mechanism for silencing antiviral immune responses and also offers new possibilities for enhancing CD8 T cell immunity in chronically infected hosts. PMID- 9858508 TI - The transcription factor early growth response 1 (Egr-1) advances differentiation of pre-B and immature B cells. AB - In mature B lymphocytes, the zinc finger transcription factor early growth response 1 (Egr-1) is one of the many immediate-early genes induced upon B cell antigen receptor engagement. However, its role during earlier stages of lymphopoiesis has remained unclear. By examining bone marrow B cell subsets, we found Egr-1 transcripts in pro/pre-B and immature B lymphocytes, and Egr-1 protein in pro/pre-B-I cells cultivated on stroma cells in the presence of interleukin (IL)-7. In recombinase-activating gene (RAG)-2-deficient mice overexpressing an Egr-1 transgene in the B lymphocyte lineage, pro/pre-B-I cells could differentiate past a developmental block at the B220(low) BP-1(-) stage to the stage of B220(low) BP-1(+) pre-B-I cells, but not further to the B220(low) BP 1(+) CD25(+) stage of pre-B-II cells. Therefore, during early B lymphopoiesis progression from the B220(low) BP-1(-) IL-2R- pro/pre-B-I stage to the B220(low) BP-1(+) IL-2R+ pre-B-II stage seems to occur in at least two distinct steps, and the first step to the stage of B220(low) BP-1(+) pre-B-I cells can be promoted by the overexpression of Egr-1 alone. Wild-type mice expressing an Egr-1 transgene had increased proportions of mature immunoglobulin (Ig)M+ B220(high) and decreased proportions of immature IgM+ B220(low) bone marrow B cells. Since transgenic and control precursor B cells show comparable proliferation patterns, overexpression of Egr-1 seems also to promote entry into the mature B cell stage. Analysis of changes in the expression pattern of potential Egr-1 target genes revealed that Egr-1 enhances the expression of the aminopeptidase BP-1/6C3 in pre B and immature B cells and upregulates expression of the orphan nuclear receptor nur77 in IgM+ B cells. PMID- 9858509 TI - Interleukin 12 and interleukin 4 control T cell adhesion to endothelial selectins through opposite effects on alpha1, 3-fucosyltransferase VII gene expression. AB - The alpha1,3-fucosyltransferase, FucT-VII, is crucial for the formation of ligands for all three selectins, and its expression regulates the synthesis of these ligands. Short-term polarized T helper (Th)1, but not Th2 or naive CD4(+) T cells, can home to sites of inflammation, but the molecular basis for this difference has remained unclear. Here we show that naive CD4(+) T cells do not express FucT-VII and fail to bind vascular selectins. We also show that when CD4(+) T cells are activated in the presence of the Th1 polarizing cytokine interleukin (IL)-12, levels of FucT-VII mRNA and binding to E- and P-selectin are significantly augmented. In contrast, activation of CD4(+) T cells in the presence of IL-4, a Th2 polarizing cytokine, inhibited FucT-VII expression and binding to vascular selectins. T cell activation upregulated expression of the Core2 transferase, C2GnT, equivalently regardless of the presence or absence of polarizing cytokines. These data indicate that the selective ability of Th1 cells, as opposed to Th2 cells or naive CD4(+) T cells, to recognize vascular selectins and home to sites of inflammation is controlled principally by the expression of a single gene, FucT-VII. PMID- 9858510 TI - Interleukin 7 receptor control of T cell receptor gamma gene rearrangement: role of receptor-associated chains and locus accessibility. AB - VDJ recombination of T cell receptor and immunoglobulin loci occurs in immature lymphoid cells. Although the molecular mechanisms of DNA cleavage and ligation have become more clear, it is not understood what controls which target loci undergo rearrangement. In interleukin 7 receptor (IL-7R)alpha-/- murine thymocytes, it has been shown that rearrangement of the T cell receptor (TCR) gamma locus is virtually abrogated, whereas other rearranging loci are less severely affected. By examining different strains of mice with targeted mutations, we now observe that the signaling pathway leading from IL-7Ralpha to rearrangement of the TCR-gamma locus requires the gammac receptor chain and the gammac-associated Janus kinase Jak3. Production of sterile transcripts from the TCR-gamma locus, a process that generally precedes rearrangement of a locus, was greatly repressed in IL-7Ralpha-/- thymocytes. The repressed transcription was not due to a lack in transcription factors since the three transcription factors known to regulate this locus were readily detected in IL-7Ralpha-/- thymocytes. Instead, the TCR-gamma locus was shown to be methylated in IL-7Ralpha-/- thymocytes. Treatment of IL-7Ralpha-/- precursor T cells with the specific histone deacetylase inhibitor trichostatin A released the block of TCR-gamma gene rearrangement. This data supports the model that IL-7R promotes TCR-gamma gene rearrangement by regulating accessibility of the locus via demethylation and histone acetylation of the locus. PMID- 9858511 TI - Genetic control of natural killing and in vivo tumor elimination by the Chok locus. AB - The molecular mechanisms underlying target recognition during natural killing are not well understood. One approach to dissect the complexities of natural killer (NK) cell recognition is through exploitation of genetic differences among inbred mouse strains. In this study, we determined that interleukin 2-activated BALB/c derived NK cells could not lyse Chinese hamster ovary (CHO) cells as efficiently as C57BL/6-derived NK cells, despite equivalent capacity to kill other targets. This strain-determined difference was also exhibited by freshly isolated NK cells, and was determined to be independent of host major histocompatibility haplotype. Furthermore, CHO killing did not correlate with expression of NK1.1 or 2B4 activation molecules. Genetic mapping studies revealed linkage between the locus influencing CHO killing, termed Chok, and loci encoded within the NK gene complex (NKC), suggesting that Chok encodes an NK cell receptor specific for CHO cells. In vivo assays recapitulated the in vitro data, and both studies determined that Chok regulates an NK perforin-dependent cytotoxic process. These results may have implications for the role of NK cells in xenograft rejection. Our genetic analysis suggests Chok is a single locus that affects NK cell mediated cytotoxicity similar to other NKC loci that also regulate the complex activity of NK cells. PMID- 9858512 TI - Complementary roles for scavenger receptor A and CD36 of human monocyte-derived macrophages in adhesion to surfaces coated with oxidized low-density lipoproteins and in secretion of H2O2. AB - Oxidized low-density lipoprotein (oxLDL) is considered one of the principal effectors of atherogenesis. To explore mechanisms by which oxLDL affects human mononuclear phagocytes, we incubated these cells in medium containing oxLDL, acetylated LDL (acLDL), or native LDL, or on surfaces coated with these native and modified lipoproteins. The presence of soluble oxLDL, acLDL, or native LDL in the medium did not stimulate H2O2 secretion by macrophages. In contrast, macrophages adherent to surfaces coated with oxLDL secreted three- to fourfold more H2O2 than macrophages adherent to surfaces coated with acLDL or native LDL. Freshly isolated blood monocytes secreted little H2O2 regardless of the substrate on which they were plated. H2O2 secretion was maximal in cells maintained for 4-6 d in culture before plating on oxLDL-coated surfaces. Fucoidan, a known ligand of class A macrophage scavenger receptors (MSR-A), significantly reduced macrophage adhesion to surfaces coated with oxLDL or acLDL. Monoclonal antibody SMO, which blocks oxLDL binding to CD36, did not inhibit adhesion of macrophages to oxLDL coated surfaces but markedly reduced H2O2 secretion by these cells. These studies show that MSR-A is primarily responsible for adhesion of macrophages to oxLDL coated surfaces, that CD36 signals H2O2 secretion by macrophages adherent to these surfaces, and that substrate-bound, but not soluble, oxLDL stimulates H2O2 secretion by macrophages. PMID- 9858513 TI - Analysis of the role of variation of major histocompatibility complex class II expression on nonobese diabetic (NOD) peripheral T cell response. AB - The current paradigm of major histocompatibility complex (MHC) and disease association suggests that efficient binding of autoantigens by disease-associated MHC molecules leads to a T cell-mediated immune response and resultant autoimmune sequelae. The data presented below offer a different model for this association of MHC with autoimmune diabetes. We used several mouse lines expressing different levels of I-Ag7 and I-Ak on the nonobese diabetic (NOD) background to evaluate the role of MHC class II in the previously described NOD T cell autoproliferation. The ratio of I-Ag7 to I-Ak expression correlated with the peripheral T cell autoproliferative phenotype in the mice studied. T cells from the NOD, [NOD x NOD. I-Anull]F1, and NOD I-Ak transgenic mice demonstrated autoproliferative responses (after priming with self-peptides), whereas the NOD.H2(h4) (containing I-Ak) congenic and [NOD x NOD. H2(h4) congenic]F1 mice did not. Analysis of CD4(+) NOD I-Ak transgenic primed lymph node cells showed that autoreactive CD4(+) T cells in the NOD I-Ak transgenic mice were restricted exclusively by I-Ag7. Considered in the context of the avidity theory of T cell activation and selection, the reported poor peptide binding capacity of NOD I-Ag7 suggested a new hypothesis to explain the effects of MHC class II expression on the peripheral autoimmune repertoire in NOD mice. This new explanation suggests that the association of MHC with diabetes results from "altered" thymic selection in which high affinity self-reactive (potentially autoreactive) T cells escape negative selection. This model offers an explanation for the requirement of homozygous MHC class II expression in NOD mice (and in humans) in susceptibility to insulin-dependent diabetes mellitus. PMID- 9858514 TI - Immunization of mice with urease vaccine affords protection against Helicobacter pylori infection in the absence of antibodies and is mediated by MHC class II restricted responses. AB - We examined the roles of cell- and antibody-mediated immunity in urease vaccine induced protection against Helicobacter pylori infection. Normal and knockout mice deficient in major histocompatibility complex (MHC) class I, MHC class II, or B cell responses were mucosally immunized with urease plus Escherichia coli heat-labile enterotoxin (LT), or parenterally immunized with urease plus aluminum hydroxide or a glycolipid adjuvant, challenged with H. pylori strain X47-2AL, and H. pylori organisms and leukocyte infiltration in the gastric mucosa quantified. In an adjuvant/route study in normal mice, there was a direct correlation between the level of protection and the density of T cells recruited to the gastric mucosa. In knockout studies, oral immunization with urease plus LT protected MHC class I knockout mice [beta2-microglobulin (-/-)] but not MHC class II knockout mice [I-Ab (-/-)]. In B cell knockout mice [microMT (-/-)], vaccine-induced protection was equivalent to that observed in immunized wild-type (+/+) mice; no IgA+ cells were detected in the stomach, but levels of CD4(+) cells equivalent to those in the wild-type strain (+/+) were seen. These studies indicate that protection of mice against H. pylori infection by immunization with the urease antigen is dependent on MHC class II-restricted, cell-mediated mechanisms, and antibody responses to urease are not required for protection. PMID- 9858515 TI - Genetic regulation of commitment to interleukin 4 production by a CD4(+) T cell intrinsic mechanism. AB - The dysregulated expression of interleukin 4 (IL-4) can have deleterious effects on the outcome of infectious and allergic diseases. Despite this, the mechanisms by which naive T cells commit to IL-4 expression during differentiation into mature effector cells remain incompletely defined. As compared to cells from most strains of mice, activated CD4(+) T cells from BALB mice show a bias towards IL-4 production and T helper 2 commitment in vitro and in vivo. Here, we show that this bias arises not from an increase in the amount of IL-4 produced per cell, but rather from an increase in the proportion of CD4(+) T cells that commit to IL 4 expression. This strain-specific difference in commitment was independent of signals mediated via the IL-4 receptor and hence occurred upstream of potential autoregulatory effects of IL-4. Segregation analysis of the phenotype in an experimental backcross cohort implicated a polymorphic locus on chromosome 16. Consistent with a role in differentiation, expression of the phenotype was CD4(+) T cell intrinsic and was evident as early as 16 h after the activation of naive T cells. Probabilistic gene activation is proposed as a T cell-intrinsic mechanism capable of modulating the proportion of naive T cells that commit to IL-4 production. PMID- 9858516 TI - CD5 expression is developmentally regulated by T cell receptor (TCR) signals and TCR avidity. AB - Recent data indicate that the cell surface glycoprotein CD5 functions as a negative regulator of T cell receptor (TCR)-mediated signaling. In this study, we examined the regulation of CD5 surface expression during normal thymocyte ontogeny and in mice with developmental and/or signal transduction defects. The results demonstrate that low level expression of CD5 on CD4(-)CD8(-) (double negative, DN) thymocytes is independent of TCR gene rearrangement; however, induction of CD5 surface expression on DN thymocytes requires engagement of the pre-TCR and is dependent upon the activity of p56(lck). At the CD4(+)CD8(+) (double positive, DP) stage, intermediate CD5 levels are maintained by low affinity TCR-major histocompatibility complex (MHC) interactions, and CD5 surface expression is proportional to both the surface level and signaling capacity of the TCR. High-level expression of CD5 on DP and CD4(+) or CD8(+) (single positive, SP) thymocytes is induced by engagement of the alpha/beta-TCR by (positively or negatively) selecting ligands. Significantly, CD5 surface expression on mature SP thymocytes and T cells was found to directly parallel the avidity or signaling intensity of the positively selecting TCR-MHC-ligand interaction. Taken together, these observations suggest that the developmental regulation of CD5 in response to TCR signaling and TCR avidity represents a mechanism for fine tuning of the TCR signaling response. PMID- 9858517 TI - Complement-dependent clearance of apoptotic cells by human macrophages. AB - Apoptotic cells are rapidly engulfed by phagocytes, but the receptors and ligands responsible for this phenomenon are incompletely characterized. Previously described receptors on blood- derived macrophages have been characterized in the absence of serum and show a relatively low uptake of apoptotic cells. Addition of serum to the phagocytosis assays increased the uptake of apoptotic cells by more than threefold. The serum factors responsible for enhanced uptake were identified as complement components that required activation of both the classical pathway and alternative pathway amplification loop. Exposure of phosphatidylserine on the apoptotic cell surface was partially responsible for complement activation and resulted in coating the apoptotic cell surface with C3bi. In the presence of serum, the macrophage receptors for C3bi, CR3 (CD11b/CD18) and CR4 (CD11c/CD18), were significantly more efficient in the uptake of apoptotic cells compared with previously described receptors implicated in clearance. Complement activation is likely to be required for efficient uptake of apoptotic cells within the systemic circulation, and early component deficiencies could predispose to systemic autoimmunity by enhanced exposure to and/or aberrant deposition of apoptotic cells. PMID- 9858518 TI - Visualization of CD4/CD8 T cell commitment. AB - A system to innocuously visualize T cell lineage commitment is described. Using a "knock-in" approach, we have generated mice expressing a beta-galactosidase reporter in place of CD4; expression of beta-galactosidase in these animals appears to be an accurate and early indicator of CD4 gene transcription. We have exploited this knock-in line to trace CD4/CD8 lineage commitment in the thymus, avoiding important pitfalls of past experimental approaches. Our results argue in favor of a selective model of thymocyte commitment, demonstrating a fundamentally symmetrical process: engagement of either class of major histocompatibility complex (MHC) molecule by a differentiating CD4(+)CD8(+) cell can give rise to T cell antigen receptor (TCR)hi thymocytes of either lineage. Key findings include (a) direct demonstration of a substantial number of CD4-committed, receptor/coreceptor-mismatched cells in MHC class II- deficient mice, a critical prediction of the selective model; (b) highly efficient rescue of such "mismatched" intermediates by forced expression of CD8 in a TCR transgenic line, and an explanation of why previous experiments of this nature were less successful-a major past criticism of the selective model; (c) direct demonstration of an analogous, though smaller, population of CD8-committed mismatched intermediates in class I-deficient animals. Finally, we found no evidence of a CD4 default pathway. PMID- 9858520 TI - The c-kit ligand, stem cell factor, can enhance innate immunity through effects on mast cells. AB - Mast cells are thought to contribute significantly to the pathology and mortality associated with anaphylaxis and other allergic disorders. However, studies using genetically mast cell-deficient WBB6F1-KitW/KitW-v and congenic wild-type (WBB6F1 +/+) mice indicate that mast cells can also promote health, by participating in natural immune responses to bacterial infection. We previously reported that repetitive administration of the c-kit ligand, stem cell factor (SCF), can increase mast cell numbers in normal mice in vivo. In vitro studies have indicated that SCF can also modulate mast cell effector function. We now report that treatment with SCF can significantly improve the survival of normal C57BL/6 mice in a model of acute bacterial peritonitis, cecal ligation and puncture (CLP). Experiments in mast cell-reconstituted WBB6F1-KitW/KitW-v mice indicate that this effect of SCF treatment reflects, at least in part, the actions of SCF on mast cells. Repetitive administration of SCF also can enhance survival in mice that genetically lack tumor necrosis factor (TNF)-alpha, demonstrating that the ability of SCF treatment to improve survival after CLP does not solely reflect effects of SCF on mast cell- dependent (or -independent) production of TNF-alpha. These findings identify c-kit and mast cells as potential therapeutic targets for enhancing innate immune responses. PMID- 9858519 TI - Type I interferon-mediated stimulation of T cells by CpG DNA. AB - Immunostimulatory DNA and oligodeoxynucleotides containing unmethylated CpG motifs (CpG DNA) are strongly stimulatory for B cells and antigen-presenting cells (APCs). We report here that, as manifested by CD69 and B7-2 upregulation, CpG DNA also induces partial activation of T cells, including naive-phenotype T cells, both in vivo and in vitro. Under in vitro conditions, CpG DNA caused activation of T cells in spleen cell suspensions but failed to stimulate highly purified T cells unless these cells were supplemented with APCs. Three lines of evidence suggested that APC-dependent stimulation of T cells by CpG DNA was mediated by type I interferons (IFN-I). First, T cell activation by CpG DNA was undetectable in IFN-IR-/- mice. Second, in contrast to normal T cells, the failure of purified IFN-IR-/- T cells to respond to CpG DNA could not be overcome by adding normal IFN-IR+ APCs. Third, IFN-I (but not IFN-gamma) caused the same pattern of partial T cell activation as CpG DNA. Significantly, T cell activation by IFN-I was APC independent. Thus, CpG DNA appeared to stimulate T cells by inducing APCs to synthesize IFN-I, which then acted directly on T cells via IFN IR. Functional studies suggested that activation of T cells by IFN-I was inhibitory. Thus, exposing normal (but not IFN-IR-/-) T cells to CpG DNA in vivo led to reduced T proliferative responses after TCR ligation in vitro. PMID- 9858521 TI - Vasostatin, a calreticulin fragment, inhibits angiogenesis and suppresses tumor growth. AB - An endothelial cell inhibitor was purified from supernatant of an Epstein-Barr virus-immortalized cell line and identified as fragments of calreticulin. The purified recombinant NH2-terminal domain of calreticulin (amino acids 1-180) inhibited the proliferation of endothelial cells, but not cells of other lineages, and suppressed angiogenesis in vivo. We have named this NH2-terminal domain of calreticulin vasostatin. When inoculated into athymic mice, vasostatin significantly reduced growth of human Burkitt lymphoma and human colon carcinoma. Compared with other inhibitors of angiogenesis, vasostatin is a small, soluble, and stable molecule that is easy to produce and deliver. As an angiogenesis inhibitor that specifically targets proliferating endothelial cells, vasostatin has a unique potential for cancer treatment. PMID- 9858522 TI - The central role of CD4(+) T cells in the antitumor immune response. AB - The induction of optimal systemic antitumor immunity involves the priming of both CD4(+) and CD8(+) T cells specific for tumor-associated antigens. The role of CD4(+) T helper cells (Th) in this response has been largely attributed to providing regulatory signals required for the priming of major histocompatibility complex class I restricted CD8(+) cytolytic T lymphocytes, which are thought to serve as the dominant effector cell mediating tumor killing. However, analysis of the effector phase of tumor rejection induced by vaccination with irradiated tumor cells transduced to secrete granulocyte/macrophage colony-stimulating factor indicates a far broader role for CD4(+) T cells in orchestrating the host response to tumor. This form of immunization leads to the simultaneous induction of Th1 and Th2 responses, both of which are required for maximal systemic antitumor immunity. Cytokines produced by these CD4(+) T cells activate eosinophils as well as macrophages that produce both superoxide and nitric oxide. Both of these cell types then collaborate within the site of tumor challenge to cause its destruction. PMID- 9858523 TI - Processing of switch transcripts is required for targeting of antibody class switch recombination. AB - Antibody class switching is mediated by somatic recombination between switch regions of the immunoglobulin heavy chain gene locus. Targeting of recombination to particular switch regions is strictly regulated by cytokines through the induction of switch transcripts starting 5' of the repetitive switch regions. However, switch transcription as such is not sufficient to target switch recombination. This has been shown in mutant mice, in which the I-exon and its promoter upstream of the switch region were replaced with heterologous promoters. Here we show that, in the murine germline targeted replacement of the endogenous gamma1 promoter, I-exon, and I-exon splice donor site by heterologous promoter and splice donor sites directs switch recombination in activated B lymphocytes constitutively to the gamma1 switch region. In contrast, switch recombination to IgG1 is inhibited in mutant mice, in which the replacement does not include the heterologous splice donor site. Our data unequivocally demonstrate that targeting of switch recombination to IgG1 in vivo requires processing of the Igamma1 switch transcripts. Either the processing machinery or the processed transcripts are involved in class switch recombination. PMID- 9858524 TI - Natural killer (NK) cell-mediated cytotoxicity: differential use of TRAIL and Fas ligand by immature and mature primary human NK cells. AB - Mature natural killer (NK) cells use Ca2+-dependent granule exocytosis and release of cytotoxic proteins, Fas ligand (FasL), and membrane-bound or secreted cytokines (tumor necrosis factor [TNF]-alpha) to induce target cell death. Fas belongs to the TNF receptor family of molecules, containing a conserved intracytoplasmic "death domain" that indirectly activates the caspase enzymatic cascade and ultimately apoptotic mechanisms in numerous cell types. Two additional members of this family, DR4 and DR5, transduce apoptotic signals upon binding soluble TNF-related apoptosis-inducing ligand (TRAIL) that, like FasL, belongs to the growing TNF family of molecules. Here, we report that TRAIL produced or expressed by different populations of primary human NK cells is functional, and represents a marker of differentiation or activation of these, and possibly other, cytotoxic leukocytes. During differentiation NK cells, sequentially and differentially, use distinct members of the TNF family or granule exocytosis to mediate target cell death. Phenotypically immature CD161(+)/CD56(-) NK cells mediate TRAIL-dependent but not FasL- or granule release-dependent cytotoxicity, whereas mature CD56(+) NK cells mediate the latter two. PMID- 9858525 TI - A critical role of natural immunoglobulin M in immediate defense against systemic bacterial infection. AB - To evaluate the role of natural immunoglobulin (Ig)M in the immediate response against microbial infection, we tested mutant mice that are deficient in secreted (s)IgM in an acute peritonitis model induced by cecal ligation and puncture (CLP). 20% of wild-type mice died within 32 h of CLP, whereas 70% of sIgM deficient mice died within the same time period. The increased susceptibility was associated with a reduced level of tumor necrosis factor (TNF)-alpha, a decreased neutrophil recruitment and an increased bacterial load in the peritoneum, and elevated levels of endotoxin and proinflammatory cytokines in the circulation. Resistance to CLP by sIgM-deficient mice was restored by reconstitution with polyclonal IgM from normal mouse serum. Reconstitution with a monoclonal IgM specific to phosphatidylcholine, a conserved cell membrane component, has a modest effect but a monoclonal IgM specific to phosphocholine is not protective. These findings demonstrate a critical role of natural IgM in the immediate defense against severe bacterial infection. PMID- 9858527 TI - p53-mediated regulation of proliferating cell nuclear antigen expression in cells exposed to ionizing radiation. AB - The proliferating cell nuclear antigen (PCNA) is a highly conserved cellular protein that functions both in DNA replication and in DNA repair. Exposure of a rat embryo fibroblast cell line (CREF cells) to gamma radiation induced simultaneous expression of PCNA with the p53 tumor suppressor protein and the cyclin-dependent kinase inhibitor p21(WAF1/Cip1). PCNA mRNA levels transiently increased in serum-starved cells exposed to ionizing radiation, an observation suggesting that the radiation-associated increase in PCNA expression could be dissociated from cell cycle progression. Irradiation of CREF cells activated a transiently expressed PCNA promoter chloramphenicol acetyltransferase construct through p53 binding sequences via a mechanism blocked by a dominant negative mutant p53. Electrophoretic mobility shift assays with nuclear extracts prepared from irradiated CREF cells produced four p53-specific DNA-protein complexes with the PCNA p53 binding site. Addition of monoclonal antibody PAb421 (p53-specific) or AC238 (specific to the transcriptional coactivator p300/CREB binding protein) to the mobility shift assay distinguished different forms of p53 that changed in relative abundance with time after irradiation. These findings suggest a complex cellular response to DNA damage in which p53 transiently activates expression of PCNA for the purpose of limited DNA repair. In a population of nongrowing cells with diminished PCNA levels, this pathway may be crucial to survival following DNA damage. PMID- 9858526 TI - c-Myc target genes involved in cell growth, apoptosis, and metabolism. PMID- 9858528 TI - Regulation of the MEF2 family of transcription factors by p38. AB - Members of the MEF2 family of transcription factors bind as homo- and heterodimers to the MEF2 site found in the promoter regions of numerous muscle specific, growth- or stress-induced genes. We showed previously that the transactivation activity of MEF2C is stimulated by p38 mitogen-activated protein (MAP) kinase. In this study, we examined the potential role of the p38 MAP kinase pathway in regulating the other MEF2 family members. We found that MEF2A, but not MEF2B or MEF2D, is a substrate for p38. Among the four p38 group members, p38 is the most potent kinase for MEF2A. Threonines 312 and 319 within the transcription activation domain of MEF2A are the regulatory sites phosphorylated by p38. Phosphorylation of MEF2A in a MEF2A-MEF2D heterodimer enhances MEF2-dependent gene expression. These results demonstrate that the MAP kinase signaling pathway can discriminate between different MEF2 isoforms and can regulate MEF2-dependent genes through posttranslational activation of preexisting MEF2 protein. PMID- 9858529 TI - The yeast telomere length counting machinery is sensitive to sequences at the telomere-nontelomere junction. AB - Saccharomyces cerevisiae telomeres consist of a continuous 325 +/- 75-bp tract of the heterogeneous repeat TG1-3 which contains irregularly spaced, high-affinity sites for the protein Rap1p. Yeast cells monitor or count the number of telomeric Rap1p molecules in a negative feedback mechanism which modulates telomere length. To investigate the mechanism by which Rap1p molecules are counted, the continuous telomeric TG1-3 sequences were divided into internal TG1-3 sequences and a terminal tract separated by nontelomeric spacers of different lengths. While all of the internal sequences were counted as part of the terminal tract across a 38 bp spacer, a 138-bp disruption completely prevented the internal TG1-3 sequences from being considered part of the telomere and defined the terminal tract as a discrete entity separate from the subtelomeric sequences. We also used regularly spaced arrays of six Rap1p sites internal to the terminal TG1-3 repeats to show that each Rap1p molecule was counted as about 19 bp of TG1-3 in vivo and that cells could count Rap1p molecules with different spacings between tandem sites. As previous in vitro experiments had shown that telomeric Rap1p sites occur about once every 18 bp, all Rap1p molecules at the junction of telomeric and nontelomeric chromatin (the telomere-nontelomere junction) must participate in telomere length measurement. The conserved arrangement of these six Rap1p molecules at the telomere-nontelomere junction in independent transformants also caused the elongated TG1-3 tracts to be maintained at nearly identical lengths, showing that sequences at the telomere-nontelomere junction had an effect on length regulation. These results can be explained by a model in which telomeres beyond a threshold length form a folded structure that links the chromosome terminus to the telomere-nontelomere junction and prevents telomere elongation. PMID- 9858530 TI - Evidence that protein binding specifies sites of DNA demethylation. AB - It has been hypothesized that protein factors may protect CpG islands from methyltransferase during development and that demethylation may involve protein DNA interactions at demethylated sites. However, direct evidence has been lacking. In this study, demethylation at the EBNA-1 binding sites of the Epstein Barr virus latent replication origin, oriP, was investigated by using human cells. Several novel findings are discussed. First, there are specific preferential demethylation sites within the oriP region. Second, the DNA sequence of oriP alone is not the target of an active demethylation process. Third, EBNA-1 binding is required for the site-specific demethylation in oriP. Interestingly, CpG sites adjacent to and between the EBNA-1 sites do not become demethylated. Fourth, demethylation of the first DNA strand in oriP at the EBNA-1 binding sites involves a passive (replication-dependent) mechanism. The second-strand demethylation appears to occur through an active mechanism. That is, EBNA-1 protein binding prevents the EBNA-1 binding sites from being remethylated after one round of DNA replication, and it appears that an active demethylase then demethylates these hemimethylated sites. This study provides clear evidence that protein binding specifies sites of DNA demethylation and provides insights into the sequence of steps and the mechanism of demethylation. PMID- 9858531 TI - RING1 interacts with multiple Polycomb-group proteins and displays tumorigenic activity. AB - Polycomb-group (PcG) proteins form large multimeric protein complexes that are involved in maintaining the transcriptionally repressive state of genes. Previously, we reported that RING1 interacts with vertebrate Polycomb (Pc) homologs and is associated with or is part of a human PcG complex. However, very little is known about the role of RING1 as a component of the PcG complex. Here we undertake a detailed characterization of RING1 protein-protein interactions. By using directed two-hybrid and in vitro protein-protein analyses, we demonstrate that RING1, besides interacting with the human Pc homolog HPC2, can also interact with itself and with the vertebrate PcG protein BMI1. Distinct domains in the RING1 protein are involved in the self-association and in the interaction with BMI1. Further, we find that the BMI1 protein can also interact with itself. To better understand the role of RING1 in regulating gene expression, we overexpressed the protein in mammalian cells and analyzed differences in gene expression levels. This analysis shows that overexpression of RING1 strongly represses En-2, a mammalian homolog of the well-characterized Drosophila PcG target gene engrailed. Furthermore, RING1 overexpression results in enhanced expression of the proto-oncogenes c-jun and c-fos. The changes in expression levels of these proto-oncogenes are accompanied by cellular transformation, as judged by anchorage-independent growth and the induction of tumors in athymic mice. Our data demonstrate that RING1 interacts with multiple human PcG proteins, indicating an important role for RING1 in the PcG complex. Further, deregulation of RING1 expression leads to oncogenic transformation by deregulation of the expression levels of certain oncogenes. PMID- 9858532 TI - hnRNP H is a component of a splicing enhancer complex that activates a c-src alternative exon in neuronal cells. AB - The regulation of the c-src N1 exon is mediated by an intronic splicing enhancer downstream of the N1 5' splice site. Previous experiments showed that a set of proteins assembles onto the most conserved core of this enhancer sequence specifically in neuronal WERI-1 cell extracts. The most prominent components of this enhancer complex are the proteins hnRNP F, KSRP, and an unidentified protein of 58 kDa (p58). This p58 protein was purified from the WERI-1 cell nuclear extract by ammonium sulfate precipitation, Mono Q chromatography, and immunoprecipitation with anti-Sm antibody Y12. Peptide sequence analysis of purified p58 protein identified it as hnRNP H. Immunoprecipitation of hnRNP H cross-linked to the N1 enhancer RNA, as well as gel mobility shift analysis of the enhancer complex in the presence of hnRNP H-specific antibodies, confirmed that hnRNP H is a protein component of the splicing enhancer complex. Immunoprecipitation of splicing intermediates from in vitro splicing reactions with anti-hnRNP H antibody indicated that hnRNP H remains bound to the src pre mRNA after the assembly of spliceosome. Partial immunodepletion of hnRNP H from the nuclear extract partially inactivated the splicing of the N1 exon in vitro. This inhibition of splicing can be restored by the addition of recombinant hnRNP H, indicating that hnRNP H is an important factor for N1 splicing. Finally, in vitro binding assays demonstrate that hnRNP H can interact with the related protein hnRNP F, suggesting that hnRNPs H and F may exist as a heterodimer in a single enhancer complex. These two proteins presumably cooperate with each other and with other enhancer complex proteins to direct splicing to the N1 exon upstream. PMID- 9858533 TI - Polypyrimidine tract-binding protein positively regulates inclusion of an alternative 3'-terminal exon. AB - Polypyrimidine tract-binding protein (PTB) is an abundant vertebrate hnRNP protein. PTB binding sites have been found within introns both upstream and downstream of alternative exons in a number of genes that are negatively controlled by the binding of PTB. We have previously reported that PTB binds to a pyrimidine tract within an RNA processing enhancer located adjacent to an alternative 3'-terminal exon within the gene coding for calcitonin and calcitonin gene-related peptide. The enhancer consists of a pyrimidine tract and CAG directly abutting on a 5' splice site sequence to form a pseudoexon. Here we show that the binding of PTB to the enhancer pyrimidine tract is functional in that exon inclusion increases when in vivo levels of PTB increase. This is the first example of positive regulation of exon inclusion by PTB. The binding of PTB was antagonistic to the binding of U2AF to the enhancer-located pyrimidine tract. Altering the enhancer pyrimidine tract to a consensus sequence for the binding of U2AF eliminated enhancement of exon inclusion in vivo and exon polyadenylation in vitro. An additional PTB binding site was identified close to the AAUAAA hexanucleotide sequence of the exon 4 poly(A) site. These observations suggest a dual role for PTB in facilitating recognition of exon 4: binding to the enhancer pyrimidine tract to interrupt productive recognition of the enhancer pseudoexon by splicing factors and interacting with the poly(A) site to positively affect polyadenylation. PMID- 9858534 TI - Functional organization of the yeast SAGA complex: distinct components involved in structural integrity, nucleosome acetylation, and TATA-binding protein interaction. AB - SAGA, a recently described protein complex in Saccharomyces cerevisiae, is important for transcription in vivo and possesses histone acetylation function. Here we report both biochemical and genetic analyses of members of three classes of transcription regulatory factors contained within the SAGA complex. We demonstrate a correlation between the phenotypic severity of SAGA mutants and SAGA structural integrity. Specifically, null mutations in the Gcn5/Ada2/Ada3 or Spt3/Spt8 classes cause moderate phenotypes and subtle structural alterations, while mutations in a third subgroup, Spt7/Spt20, as well as Ada1, disrupt the complex and cause severe phenotypes. Interestingly, double mutants (gcn5Delta spt3Delta and gcn5Delta spt8Delta) causing loss of a member of each of the moderate classes have severe phenotypes, similar to spt7Delta, spt20Delta, or ada1Delta mutants. In addition, we have investigated biochemical functions suggested by the moderate phenotypic classes and find that first, normal nucleosomal acetylation by SAGA requires a specific domain of Gcn5, termed the bromodomain. Deletion of this domain also causes specific transcriptional defects at the HIS3 promoter in vivo. Second, SAGA interacts with TBP, the TATA-binding protein, and this interaction requires Spt8 in vitro. Overall, our data demonstrate that SAGA harbors multiple, distinct transcription-related functions, including direct TBP interaction and nucleosomal histone acetylation. Loss of either of these causes slight impairment in vivo, but loss of both is highly detrimental to growth and transcription. PMID- 9858535 TI - AnCF, the CCAAT binding complex of Aspergillus nidulans, contains products of the hapB, hapC, and hapE genes and is required for activation by the pathway-specific regulatory gene amdR. AB - CCAAT binding factors (CBFs) positively regulating the expression of the amdS gene (encoding acetamidase) and two penicillin biosynthesis genes (ipnA and aatA) have been previously found in Aspergillus nidulans. The factors were called AnCF and PENR1, respectively. Deletion of the hapC gene, encoding a protein with significant similarity to Hap3p of Saccharomyces cerevisiae, eliminated both AnCF and PENR1 binding activities. We now report the isolation of the genes hapB and hapE, which encode proteins with central regions of high similarity to Hap2p and Hap5p of S. cerevisiae and to the CBF-B and CBF-C proteins of mammals. An additional fungus-specific domain present in HapE was revealed by comparisons with the homologs from S. cerevisiae, Neurospora crassa, and Schizosaccharomyces pombe. The HapB, HapC, and HapE proteins have been shown to be necessary and sufficient for the formation of a CCAAT binding complex in vitro. Strains with deletions of each of the hapB, hapC, and hapE genes have identical phenotypes of slow growth, poor conidiation, and reduced expression of amdS. Furthermore, induction of amdS by omega amino acids, which is mediated by the AmdR pathway specific activator, is abolished in the hap deletion mutants, as is growth on gamma-aminobutyric acid as a sole nitrogen or carbon source. AmdR and AnCF bind to overlapping sites in the promoters of the amdS and gatA genes. It is known that AnCF can bind independently of AmdR. We suggest that AnCF binding is required for AmdR binding in vivo. PMID- 9858536 TI - Mechanism of regulatory target selection by the SOX high-mobility-group domain proteins as revealed by comparison of SOX1/2/3 and SOX9. AB - SOX proteins bind similar DNA motifs through their high-mobility-group (HMG) domains, but their action is highly specific with respect to target genes and cell type. We investigated the mechanism of target selection by comparing SOX1/2/3, which activate delta-crystallin minimal enhancer DC5, with SOX9, which activates Col2a1 minimal enhancer COL2C2. These enhancers depend on both the SOX binding site and the binding site of a putative partner factor. The DC5 site was equally bound and bent by the HMG domains of SOX1/2 and SOX9. The activation domains of these SOX proteins mapped at the distal portions of the C-terminal domains were not cell specific and were independent of the partner factor. Chimeric proteins produced between SOX1 and SOX9 showed that to activate the DC5 enhancer, the C-terminal domain must be that of SOX1, although the HMG domains were replaceable. The SOX2-VP16 fusion protein, in which the activation domain of SOX2 was replaced by that of VP16, activated the DC5 enhancer still in a partner factor-dependent manner. The results argue that the proximal portion of the C terminal domain of SOX1/2 specifically interacts with the partner factor, and this interaction determines the specificity of the SOX1/2 action. Essentially the same results were obtained in the converse experiments in which COL2C2 activation by SOX9 was analyzed, except that specificity of SOX9-partner factor interaction also involved the SOX9 HMG domain. The highly selective SOX-partner factor interactions presumably stabilize the DNA binding of the SOX proteins and provide the mechanism for regulatory target selection. PMID- 9858537 TI - Spi-1/PU.1 is a positive regulator of the Fli-1 gene involved in inhibition of erythroid differentiation in friend erythroleukemic cell lines. AB - Spi-1/PU.1 and Fli-1 are two members of the ETS family of transcription factors whose expression is deregulated by proviral insertion in most erythroleukemic cell lines induced by the spleen focus-forming virus (SFFV) and Friend murine leukemia virus (F-MuLV) components of the Friend viral complex, respectively. In this study, we present evidence that transcription of the Fli-1 gene is positively regulated by Spi-1/PU.1 in SFFV-transformed cell lines: (i) all SFFV transformed cell lines expressing Spi-1/PU.1 are characterized by a specific pattern of Fli-1 gene transcripts initiated in the -200 region instead of position -400 as reported for F-MuLV-transformed cell lines; (ii) these Fli-1 transcripts initiated in the -200 region are downregulated in parallel with that of Spi-1/PU.1 during hexamethylenebisacetamide (HMBA) induced differentiation; and (iii) Fli-1 transcription is upregulated in SFFV cells lines following stable transfection of a Spi-1/PU.1 expression vector. Furthermore, we found by transient transfection assays that the -270/-41 region of the Fli-1 gene displays promoter activity which is transactivated by Spi-1/PU.1. This promoter is strictly dependent on the integrity of two highly conserved ETS DNA binding sites that bind the Spi-1/PU.1 protein in vitro. Finally, we show that transfection of constitutive or inducible Fli-1 expression vectors in SFFV-transformed cells inhibits their erythroid differentiation induced by HMBA. Overall, these data indicate that Fli-1 is a target gene of the Spi-1/PU.1 transcription factor in SFFV-transformed cell lines. We further suggest that deregulated synthesis of Fli 1 may trigger a common mechanism contributing to erythroleukemia induced by either SFFV or F-MuLV. PMID- 9858538 TI - Glutamate induces phosphorylation of Elk-1 and CREB, along with c-fos activation, via an extracellular signal-regulated kinase-dependent pathway in brain slices. AB - In cell culture systems, the TCF Elk-1 represents a convergence point for extracellular signal-related kinase (ERK) and c-Jun N-terminal kinase/stress activated protein kinase (JNK/SAPK) subclasses of mitogen-activated protein kinase (MAPK) cascades. Its phosphorylation strongly potentiates its ability to activate transcription of the c-fos promoter through a ternary complex assembled on the c-fos serum response element. In rat brain postmitotic neurons, Elk-1 is strongly expressed (V. Sgambato, P. Vanhoutte, C. Pages, M. Rogard, R. A. Hipskind, M. J. Besson, and J. Caboche, J. Neurosci. 18:214-226, 1998). However, its physiological role in these postmitotic neurons remains to be established. To investigate biochemically the signaling pathways targeting Elk-1 and c-fos in mature neurons, we used a semi-in vivo system composed of brain slices stimulated with the excitatory neurotransmitter glutamate. Glutamate treatment leads to a robust, progressive activation of the ERK and JNK/SAPK MAPK cascades. This corresponds kinetically to a significant increase in Ser383-phosphorylated Elk-1 and the appearance of c-fos mRNA. Glutamate also causes increased levels of Ser133-phosphorylated cyclic AMP-responsive element-binding protein (CREB) but only transiently relative to Elk-1 and c-fos. ERK and Elk-1 phosphorylation are blocked by the MAPK kinase inhibitor PD98059, indicating the primary role of the ERK cascade in mediating glutamate signaling to Elk-1 in the rat striatum in vivo. Glutamate-mediated CREB phosphorylation is also inhibited by PD98059 treatment. Interestingly, KN62, which interferes with calcium-calmodulin kinase (CaM-K) activity, leads to a reduction of glutamate-induced ERK activation and of CREB phosphorylation. These data indicate that ERK functions as a common component in two signaling pathways (ERK/Elk-1 and ERK/?/CREB) converging on the c-fos promoter in postmitotic neuronal cells and that CaM-Ks act as positive regulators of these pathways. PMID- 9858539 TI - Abnormal, error-prone bypass of photoproducts by xeroderma pigmentosum variant cell extracts results in extreme strand bias for the kinds of mutations induced by UV light. AB - Xeroderma pigmentosum (XP) is a rare genetic disease characterized by a greatly increased susceptibility to sunlight-induced skin cancer. Cells from the majority of patients are defective in nucleotide excision repair. However, cells from one set of patients, XP variants, exhibit normal repair but are abnormally slow in replicating DNA containing UV photoproducts. The frequency of UV radiation induced mutations in the XP variant cells is significantly higher than that in normal human cells. Furthermore, the kinds of UV-induced mutations differ very significantly from normal. Instead of transitions, mainly C-->T, 30% of the base substitutions consist of C-->A transversions, all arising from photoproducts located in one strand. Mutations involving cytosine in the other strand are almost all C-->T transitions. Forty-five percent of the substitutions involve thymine, and the majority are transversions. To test the hypothesis that the UV hypermutability and the abnormal spectrum of mutations result from abnormal bypass of photoproducts in DNA, we compared extracts from XP variant cells with those from HeLa cells and a fibroblast cell strain, MSU-1.2, for the ability to replicate a UV-irradiated form I M13 phage. The M13 template contains a simian virus 40 origin of replication located directly to the left or to the right of the target gene, lacZalpha, so that the template for the leading and lagging strands of DNA replication is defined. Reduction of replication to approximately 37% of the control value required only 1 photoproduct per template for XP variant cell extracts, but approximately 2.2 photoproducts for HeLa or MSU-1.2 cell extracts. The frequency of mutants induced was four times higher with XP variant cell extracts than with HeLa or MSU-1.2 cell extracts. With XP variant cell extracts, the proportion of C-->A transversions reached as high as 43% with either M13 template and arose from photoproducts located in the template for leading-strand synthesis; with HeLa or MSU-1.2 cell extracts, this value was only 5%, and these arose from photoproducts in either strand. With the XP variant extracts, 26% of the substitutions involved thymine, and virtually all were T-->A transversions. Sequence analysis of the coding region of the catalytic subunit of DNA polymerase delta in XP variant cell lines revealed two polymorphisms, but these do not account for the reduced bypass fidelity. Our data indicate that the UV hypermutability of XP variant cells results from reduced bypass fidelity and that unlike for normal cells, bypass of photoproducts involving cytosine in the template for the leading strand differs significantly from that of photoproducts in the lagging strand. PMID- 9858540 TI - A novel intron element operates posttranscriptionally To regulate human N-myc expression. AB - Precisely regulated expression of oncogenes and tumor suppressor genes is essential for normal development, and deregulated expression can lead to cancer. The human N-myc gene normally is expressed in only a subset of fetal epithelial tissues, and its expression is extinguished in all adult tissues except transiently in pre-B lymphocytes. The N-myc gene is overexpressed due to genomic amplification in the childhood tumor neuroblastoma. In previous work to investigate mechanisms of regulation of human N-myc gene expression, we observed that N-myc promoter-chloramphemicol acelyltransferase reporter constructs containing sequences 5' to exon 1 were active in all cell types examined, regardless of whether endogenous N-myc RNA was detected. In contrast, inclusion of the first exon and a portion of the first intron allowed expression only in those cell types with detectable endogenous N-myc transcripts. We investigated further the mechanisms by which this tissue-specific control of N-myc expression is achieved. Using nuclear run-on analyses, we determined that the N-myc gene is actively transcribed in all cell types examined, indicating a posttranscriptional mode of regulation. Using a series of N-myc intron 1 deletion constructs, we localized a 116-bp element (tissue-specific element [TSE]) within the first intron that directs tissue-specific N-myc expression. The TSE can function independently to regulate expression of a heterologous promoter-reporter minigene in a cell-specific pattern that mirrors the expression pattern of the endogenous N-myc gene. Surprisingly, the TSE can function in both sense and antisense orientations to regulate gene expression. Our data indicate that the human N-myc TSE functions through a posttranscriptional mechanism to regulate N-myc expression. PMID- 9858541 TI - DNA methylation profile of the mouse skeletal alpha-actin promoter during development and differentiation. AB - Genomic levels of DNA methylation undergo widespread alterations in early embryonic development. However, changes in embryonic methylation have proven difficult to study at the level of single-copy genes due to the small amount of tissue available for assay. This study provides the first detailed analysis of the methylation state of a tissue-specific gene through early development and differentiation. Using bisulfite sequencing, we mapped the methylation profile of the tissue-specific mouse skeletal alpha-actin promoter at all stages of development, from gametes to postimplantation embryos. We show that the alpha actin promoter, which is fully methylated in the sperm and essentially unmethylated in the oocyte, undergoes a general demethylation from morula to blastocyst stages, although the blastula is not completely demethylated. Remethylation of the alpha-actin promoter occurs after implantation in a stochastic pattern, with some molecules being extensively methylated and others sparsely methylated. Moreover, we demonstrate that tissue-specific expression of the skeletal alpha-actin gene in the adult mouse does not correlate with the methylation state of the promoter, as we find a similar low level of methylation in both expressing and one of the two nonexpressing tissues tested. However, a subset of CpG sites within the skeletal alpha-actin promoter are preferentially methylated in liver, a nonexpressing tissue. PMID- 9858542 TI - The beta subunit of eukaryotic translation initiation factor 2 binds mRNA through the lysine repeats and a region comprising the C2-C2 motif. AB - Eukaryotic translation initiation factor 2 (eIF2) has been implicated in the selection of the AUG codon as the start site for eukaryotic translation initiation, since mutations in its three subunits in yeast that allow the recognition of a UUG codon by the anticodon of the initiator Met-tRNAMet have been identified. All such mutations in the beta subunit of eIF2 (eIF2beta) mapped to a region containing a putative zinc finger structure of the C2-C2 type, indicating that these sequences could be involved in RNA recognition. Another feature of eIF2beta that could mediate an interaction with RNA is located in the amino-terminal sequences and is composed of three repeats of seven lysine residues which are highly conserved in other species. We show here the ability of eIF2beta, purified from Escherichia coli as a fusion to glutathione S transferase, to bind mRNA in vitro. Through a deletion analysis, mRNA binding was found to be dependent on the lysine repeats and a region encompassing the C2-C2 motif. Strong mRNA binding in vitro could be maintained by the presence of only one lysine or one arginine run but not one alanine run. We further show that only one run of lysine residues is sufficient for the in vivo function of eIF2beta, probably through charge interaction, since its replacement by arginines did not impair cell viability, whereas substitution for alanines resulted in inviable cells. mRNA binding, but not GTP-dependent initiator Met-tRNAMet binding, by the eIF2 complex was determined to be dependent on the presence of the lysine runs of the beta subunit. PMID- 9858543 TI - Alternative splicing results in differential expression, activity, and localization of the two forms of arginyl-tRNA-protein transferase, a component of the N-end rule pathway. AB - The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. The underlying ubiquitin-dependent proteolytic system, called the N-end rule pathway, is organized hierarchically: N-terminal aspartate and glutamate (and also cysteine in metazoans) are secondary destabilizing residues, in that they function through their conjugation, by arginyl-tRNA-protein transferase (R-transferase), to arginine, a primary destabilizing residue. We isolated cDNA encoding the 516-residue mouse R-transferase, ATE1p, and found two species, termed Ate1-1 and Ate1-2. The Ate1 mRNAs are produced through a most unusual alternative splicing that retains one or the other of the two homologous 129-bp exons, which are adjacent in the mouse Ate1 gene. Human ATE1 also contains the alternative 129-bp exons, whereas the plant (Arabidopsis thaliana) and fly (Drosophila melanogaster) Ate1 genes encode a single form of ATE1p. A fusion of ATE1-1p with green fluorescent protein (GFP) is present in both the nucleus and the cytosol, whereas ATE1-2p-GFP is exclusively cytosolic. Mouse ATE1-1p and ATE1 2p were examined by expressing them in ate1Delta Saccharomyces cerevisiae in the presence of test substrates that included Asp-betagal (beta-galactosidase) and Cys-betagal. Both forms of the mouse R-transferase conferred instability on Asp betagal (but not on Cys-betagal) through the arginylation of its N-terminal Asp, the ATE1-1p enzyme being more active than ATE1-2p. The ratio of Ate1-1 to Ate1-2 mRNA varies greatly among the mouse tissues; it is approximately 0.1 in the skeletal muscle, approximately 0.25 in the spleen, approximately 3.3 in the liver and brain, and approximately 10 in the testis, suggesting that the two R transferases are functionally distinct. PMID- 9858544 TI - Transcriptional regulation of the AP-2alpha promoter by BTEB-1 and AP-2rep, a novel wt-1/egr-related zinc finger repressor. AB - AP-2 transcription factors have been suggested to exert key regulatory functions in vertebrate embryonic development, in tumorigenicity of various cancer cell types, and in controlling cell cycle and apoptotic effector genes. In this study, we investigated transcriptional regulation of the AP-2alpha gene promoter mediated by an autoregulatory element (referred to as A32) with a core consensus AP-2 binding site at position -336 relative to the mRNA initiation site. AP-2 and multiple different nuclear proteins in HeLa and Neuro2A cell extracts form specific bandshifts with the A32 element. By screening a mouse brain cDNA expression library, we isolated two different cDNAs encoding the transcription factor BTEB-1 and a novel zinc finger protein, AP-2rep. AP-2rep reveals a modular structure with homology to transcription factors of the wt-1/egr-1-family. AP 2rep, BTEB-1, and AP-2 interact in a mutually exclusive manner with overlapping binding sites in the A32 element. Transfection studies revealed that BTEB-1 is a strong activator of AP-2alpha promoter activity, whereas cotransfected AP-2alpha resulted in moderate autoactivation of promoter activity. In contrast, AP-2rep confers strong transcriptional repression to the AP-2alpha gene, and we observed an excellent correlation between induction of AP-2rep mRNA expression and downregulation of AP-2alpha mRNA during development of the kidney. In summary, we have identified multiple transcription factors and cloned from an expression library a novel zinc finger silencing factor, AP-2rep, mediating positive and negative regulation of AP-2alpha expression through a set of overlapping cis regulatory promoter elements. PMID- 9858545 TI - p21(Waf1/Cip1) inhibition of cyclin E/Cdk2 activity prevents endoreduplication after mitotic spindle disruption. AB - During a normal cell cycle, entry into S phase is dependent on completion of mitosis and subsequent activation of cyclin-dependent kinases (Cdks) in G1. These events are monitored by checkpoint pathways. Recent studies and data presented herein show that after treatment with microtubule inhibitors (MTIs), cells deficient in the Cdk inhibitor p21(Waf1/Cip1) enter S phase with a >/=4N DNA content, a process known as endoreduplication, which results in polyploidy. To determine how p21 prevents MTI-induced endoreduplication, the G1/S and G2/M checkpoint pathways were examined in two isogenic cell systems: HCT116 p21(+/+) and p21(-/-) cells and H1299 cells containing an inducible p21 expression vector (HIp21). Both HCT116 p21(-/-) cells and noninduced HIp21 cells endoreduplicated after MTI treatment. Analysis of G1-phase Cdk activities demonstrated that the induction of p21 inhibited endoreduplication through direct cyclin E/Cdk2 regulation. The kinetics of p21 inhibition of cyclin E/Cdk2 activity and binding to proliferating-cell nuclear antigen in HCT116 p21(+/+) cells paralleled the onset of endoreduplication in HCT116 p21(-/-) cells. In contrast, loss of p21 did not lead to deregulated cyclin D1-dependent kinase activities, nor did p21 directly regulate cyclin B1/Cdc2 activity. Furthermore, we show that MTI-induced endoreduplication in p53-deficient HIp21 cells was due to levels of p21 protein below a threshold required for negative regulation of cyclin E/Cdk2, since ectopic expression of p21 restored cyclin E/Cdk2 regulation and prevented endoreduplication. Based on these findings, we propose that p21 plays an integral role in the checkpoint pathways that restrain normal cells from entering S phase after aberrant mitotic exit due to defects in microtubule dynamics. PMID- 9858546 TI - FLP recombinase-mediated induction of Cu/Zn-superoxide dismutase transgene expression can extend the life span of adult Drosophila melanogaster flies. AB - Yeast FLP recombinase was used in a binary transgenic system ("FLP-OUT") to allow induced overexpression of catalase and/or Cu/Zn-superoxide dismutase (Cu/ZnSOD) in adult Drosophila melanogaster. Expression of FLP recombinase was driven by the heat-inducible hsp70 promoter. Once expressed, FLP catalyzed the rearrangement and activation of a target construct in which expression of catalase or Cu/ZnSOD cDNAs was driven by the constitutive actin5C promoter. In this way a brief heat pulse (120 or 180 min, total) of young adult flies activated transgene expression for the rest of the life span. FLP-OUT allows the effects of induced transgene expression to be analyzed in control (no heat pulse) and experimental (heat pulse) populations with identical genetic backgrounds. Under the conditions used, the heat pulse itself always had neutral or slightly negative effects on the life span. Catalase overexpression significantly increased resistance to hydrogen peroxide but had neutral or slightly negative effects on the mean life span. Cu/ZnSOD overexpression extended the mean life span up to 48%. Simultaneous overexpression of catalase with Cu/ZnSOD had no added benefit, presumably due to a preexisting excess of catalase. The data suggest that oxidative damage is one rate-limiting factor for the life span of adult Drosophila. Finally, experimental manipulation of the genetic background demonstrated that the life span is affected by epistatic interactions between the transgene and allele(s) at other loci. PMID- 9858547 TI - Identification of constitutive and ras-inducible phosphorylation sites of KSR: implications for 14-3-3 binding, mitogen-activated protein kinase binding, and KSR overexpression. AB - Genetic and biochemical studies have identified kinase suppressor of Ras (KSR) to be a conserved component of Ras-dependent signaling pathways. To better understand the role of KSR in signal transduction, we have initiated studies investigating the effect of phosphorylation and protein interactions on KSR function. Here, we report the identification of five in vivo phosphorylation sites of KSR. In serum-starved cells, KSR contains two constitutive sites of phosphorylation (Ser297 and Ser392), which mediate the binding of KSR to the 14-3 3 family of proteins. In the presence of activated Ras, KSR contains three additional sites of phosphorylation (Thr260, Thr274, and Ser443), all of which match the consensus motif (Px[S/T]P) for phosphorylation by mitogen-activated protein kinase (MAPK). Further, we find that treatment of cells with the MEK inhibitor PD98059 blocks phosphorylation of the Ras-inducible sites and that activated MAPK associates with KSR in a Ras-dependent manner. Together, these findings indicate that KSR is an in vivo substrate of MAPK. Mutation of the identified phosphorylation sites did not alter the ability of KSR to facilitate Ras signaling in Xenopus oocytes, suggesting that phosphorylation at these sites may serve other functional roles, such as regulating catalytic activity. Interestingly, during the course of this study, we found that the biological effect of KSR varied dramatically with the level of KSR protein expressed. In Xenopus oocytes, KSR functioned as a positive regulator of Ras signaling when expressed at low levels, whereas at high levels of expression, KSR blocked Ras dependent signal transduction. Likewise, overexpression of Drosophila KSR blocked R7 photoreceptor formation in the Drosophila eye. Therefore, the biological function of KSR as a positive effector of Ras-dependent signaling appears to be dependent on maintaining KSR protein expression at low or near-physiological levels. PMID- 9858548 TI - The msh2 gene of Schizosaccharomyces pombe is involved in mismatch repair, mating type switching, and meiotic chromosome organization. AB - We have identified in the fission yeast Schizosaccharomyces pombe a MutS homolog that shows highest homology to the Msh2 subgroup. msh2 disruption gives rise to increased mitotic mutation rates and increased levels of postmeiotic segregation of genetic markers. In bandshift assays performed with msh2Delta cell extracts, a general mismatch-binding activity is absent. By complementation assays, we showed that S. pombe msh2 is allelic with the previously identified swi8 and mut3 genes, which are involved in mating-type switching. The swi8-137 mutant has a mutation in the msh2 gene which causes a truncated Msh2 peptide lacking a putative DNA binding domain. Cytological analysis revealed that during meiotic prophase of msh2-defective cells, chromosomal structures were frequently formed; such structures are rarely found in the wild type. Our data show that besides having a function in mismatch repair, S. pombe msh2 is required for correct termination of copy synthesis during mating-type switching as well as for proper organization of chromosomes during meiosis. PMID- 9858549 TI - hnRNP A1 recruited to an exon in vivo can function as an exon splicing silencer. AB - Some exons contain exon splicing silencers. Their activity is frequently balanced by that of splicing enhancers, and this is important to ensure correct relative levels of alternatively spliced mRNAs. Using an immunoprecipitation and UV-cross linking assay, we show that RNA molecules containing splicing silencers from the human immunodeficiency virus type 1 tat exon 2 or the human fibroblast growth factor receptor 2 K-SAM exon bind to hnRNP A1 in HeLa cell nuclear extracts better than the corresponding RNA molecule without a silencer. Two different point mutations which abolish the K-SAM exon splicing silencer's activity reduce hnRNP A1 binding twofold. Recruitment of hnRNP A1 in the form of a fusion with bacteriophage MS2 coat protein to a K-SAM exon whose exon splicing silencer has been replaced by a coat binding site efficiently represses splicing of the exon in vivo. Recruitment of only the glycine-rich C-terminal domain of hnRNP A1, which is capable of interactions with other proteins, is sufficient to repress exon splicing. Our results show that hnRNP A1 can function to repress splicing, and they suggest that at least some exon splicing silencers could work by recruiting hnRNP A1. PMID- 9858550 TI - Multiple distinct splicing enhancers in the protein-coding sequences of a constitutively spliced pre-mRNA. AB - We have identified multiple distinct splicing enhancer elements within protein coding sequences of the constitutively spliced human beta-globin pre-mRNA. Each of these highly conserved sequences is sufficient to activate the splicing of a heterologous enhancer-dependent pre-mRNA. One of these enhancers is activated by and binds to the SR protein SC35, whereas at least two others are activated by the SR protein SF2/ASF. A single base mutation within another enhancer element inactivates the enhancer but does not change the encoded amino acid. Thus, overlapping protein coding and RNA recognition elements may be coselected during evolution. These studies provide the first direct evidence that SR protein specific splicing enhancers are located within the coding regions of constitutively spliced pre-mRNAs. We propose that these enhancers function as multisite splicing enhancers to specify 3' splice-site selection. PMID- 9858551 TI - Transcriptional and posttranscriptional silencing of rodent alpha1(I) collagen by a homologous transcriptionally self-silenced transgene. AB - Transient transfection of rodent fibroblasts with plasmids carrying a full-size pro-alpha1(I) collagen gene (pWTC1) results in rapid reduction of the endogenous transcripts by >90%, while the transgene mRNA is undetectable. Using deletion constructs, two adjacent 5' noncoding regions of the gene are identified as causing transcriptional silencing of the endogene in normal and v-fos-transformed cells but not in nontumorigenic revertants, which show partial relief from v-fos transformation-induced alpha1(I) gene suppression. The 3' end of the transgene carries an additional element(s), causing posttranscriptional silencing of the endogene in all cells including the revertant. Data indicate that the transgenes are transcriptionally self-silenced. Genome-integrated transgenes that are transcriptionally active also allow expression of the endogene, suggesting gene activation by chromosomal factors missing in pWTC1. Silencing is not regulated by antisense RNA. Silencing of the endogenous pro-alpha1(I) collagen gene is not linked to the level of transgene expression. PMID- 9858552 TI - Cux/CDP homeoprotein is a component of NF-muNR and represses the immunoglobulin heavy chain intronic enhancer by antagonizing the bright transcription activator. AB - Nuclear matrix attachment regions (MARs) flanking the immunoglobulin heavy chain intronic enhancer (Emu) are the targets of the negative regulator, NF-muNR, found in non-B and early pre-B cells. Expression library screening with NF-muNR binding sites yielded a cDNA clone encoding an alternatively spliced form of the Cux/CDP homeodomain protein. Cux/CDP fulfills criteria required for NF-muNR identity. It is expressed in non-B and early pre-B cells but not mature B cells. It binds to NF-muNR binding sites within Emu with appropriate differential affinities. Antiserum specific for Cux/CDP recognizes a polypeptide of the predicted size in affinity-purified NF-muNR preparations and binds NF-muNR complexed with DNA. Cotransfection with Cux/CDP represses the activity of Emu via the MAR sequences in both B and non-B cells. Cux/CDP antagonizes the effects of the Bright transcription activator at both the DNA binding and functional levels. We propose that Cux/CDP regulates cell-type-restricted, differentiation stage-specific Emu enhancer activity by interfering with the function of nuclear matrix-bound transcription activators. PMID- 9858553 TI - Poly(ADP-ribose) polymerase binds with transcription enhancer factor 1 to MCAT1 elements to regulate muscle-specific transcription. AB - Striated muscle-specific expression of the cardiac troponin T (cTNT) gene is mediated through two MCAT elements that act via binding of transcription enhancer factor 1 (TEF-1) to the MCAT core motifs and binding of an auxiliary protein to nucleotides flanking the 5' side of the core motif. Using DNA-protein and protein protein binding experiments, we identified a 140-kDa polypeptide that bound both the muscle-specific flanking sequences of the most distal MCAT1 element and TEF 1. Screening of an expression library with the MCAT1 element yielded a cDNA encoding a truncated form of poly(ADP-ribose) polymerase (PARP). Endogenous PARP from embryonic tissue nuclear extracts migrated as a 140-kDa protein. Recombinant full-length PARP preferentially bound the wild-type MCAT1 element and was shown to physically interact with TEF-1. In addition, endogenous TEF-1 could be coimmunoprecipitated with PARP from extracts of primary skeletal muscle cells. Recombinant PARP was able to ADP-ribosylate TEF-1 in vitro. Inhibition of the enzymatic activity of PARP repressed expression of an MCAT1-dependent reporter in transiently transfected primary muscle cells. Together, these data implicate PARP as the auxiliary protein that binds with TEF-1 to the MCAT1 element to provide muscle-specific gene transcription. PMID- 9858555 TI - Deletion of 11 amino acids in p90(rsk-mo-1) abolishes kinase activity. AB - p90(rsk) is a distal member of the mitogen-activated protein kinase signaling pathway. It has been cloned from a variety of species including Xenopus laevis, mouse, chicken, rat, and human. The clone p90(rsk-mo-1), isolated by others from a mouse library, contains a unique 33-nucleotide deletion not found in the p90(rsk) clones from any other species that have been examined. When p90(rsk-mo 1) was expressed in Cos-7 cells that were subsequently stimulated with epidermal growth factor, the immunoprecipitated p90(rsk-mo-1) protein showed no measurable kinase activity toward the ribosomal protein S6 peptide. By comparison, expression of rat p90(rsk-1) resulted in significant kinase activity. Deletion of the 33-nucleotide region missing in the p90(rsk-mo-1) clone from the p90(rsk-rat 1) cDNA abolished kinase activity in the resulting protein. When these 33 nucleotides were introduced into the p90(rsk-mo-1) cDNA, the expressed protein showed significant kinase activity. Reverse transcription-PCR and direct sequencing of mRNA isolated from several mouse tissues indicated the presence of the full-length form of p90(rsk-1) in the mouse and showed no conclusive evidence for a deletion-containing form. This study indicates the presence of a full length p90(rsk-1) mRNA in mouse tissues that is homologous to that identified in other species and suggests that the deletion in p90(rsk-mo-1) may be a cloning artifact. The findings provide additional support for the conclusion that the first catalytic domain of p90(rsk) is responsible for its enzymatic activity toward ribosomal protein S6. PMID- 9858554 TI - Mapping of a serine-rich domain essential for the transcriptional, antiapoptotic, and transforming activities of the v-Rel oncoprotein. AB - The v-Rel oncoprotein belongs to the Rel/NF-kappaB family of transcription factors and induces aggressive lymphomas in chickens and transgenic mice. Current models for cell transformation by v-Rel invoke the combined activation of gene expression and the dominant inhibition of transcription mediated by its cellular homologs. Here, we mapped a serine-rich transactivation domain in the C terminus of v-Rel that is necessary for its biological activity. Specific serine-to alanine substitutions within this region impaired the transcriptional activity of v-Rel, whereas a double mutant abolished its function. In contrast, substitutions with phosphomimetic aspartate residues led to a complete recovery of the transcriptional potential. The transforming activity of v-Rel mutants correlated with their ability to inhibit programmed cell death. The transforming and antiapoptotic activities of v-Rel were abolished by defined Ser-to-Ala mutations and restored by most Ser-to-Asp substitutions. However, one Ser-to-Asp mutant showed wild-type transactivation ability but failed to block apoptosis and to transform cells. These results show that the transactivation function of v-Rel is necessary but not sufficient for cell transformation, adding an important dimension to the transformation model. It is possible that defined protein protein interactions are also required to block apoptosis and transform cells. Since v-Rel is an acutely oncogenic member of the Rel/NF-kappaB family, our data raise the possibility that phosphorylation of its serine-rich transactivation domain may regulate its unique biological activity. PMID- 9858556 TI - Activated MEK stimulates expression of AP-1 components independently of phosphatidylinositol 3-kinase (PI3-kinase) but requires a PI3-kinase signal To stimulate DNA synthesis. AB - To investigate the contribution that ERK/mitogen-activated protein kinase signalling makes to cell cycle progression and gene expression, we have constructed cell lines to express an inducible version of activated MEK1. Using these cells, we show that activation of MEK leads to the expression of Fra-1 and Fra-2 but not c-Fos. Treatment of Ras-transformed cells with the MEK inhibitor PD098059 blocks expression of Fra-1 and Fra-2, showing that in Ras transformation ERK signalling is responsible for Fra-1 and Fra-2 expression. Activation of MEK1 in growth-arrested cells leads to DNA synthesis; however, ERK activation alone is insufficient because the induction of DNA synthesis is blocked by inhibition of phosphatidylinositol 3-kinase (PI3-kinase). Activation of PI3-kinase is indirect, perhaps through autocrine growth factors, and is required for the induction of cyclin D1. PMID- 9858558 TI - Functional domains of the Rsp5 ubiquitin-protein ligase. AB - RSP5, an essential gene of Saccharomyces cerevisiae, encodes a hect domain E3 ubiquitin-protein ligase. Hect E3 proteins have been proposed to consist of two broad functional domains: a conserved catalytic carboxyl-terminal domain of approximately 350 amino acids (the hect domain) and a large, nonconserved amino terminal domain containing determinants of substrate specificity. We report here the mapping of the minimal region of Rsp5 necessary for its essential in vivo function, the minimal region necessary to stably interact with a substrate of Rsp5 (Rpb1, the large subunit of RNA polymerase II), and the finding that the hect domain, by itself, is sufficient for formation of the ubiquitin-thioester intermediate. Mutations within the hect domain that affect either the ability to form a ubiquitin-thioester or to catalyze substrate ubiquitination abrogate in vivo function, strongly suggesting that the ubiquitin-protein ligase activity of Rsp5 is intrinsically linked to its essential function. The amino-terminal region of Rsp5 contains three WW domains and a C2 calcium-binding domain. Two of the three WW domains are required for the essential in vivo function, while the C2 domain is not, and requirements for Rpb1 binding and ubiquitination lie within the region required for in vivo function. Together, these results support the two domain model for hect E3 function and indicate that the WW domains play a role in the recognition of at least some of the substrates of Rsp5, including those related to its essential function. In addition, we show that haploid yeast strains bearing complete disruptions of either of two other hect E3 genes of yeast, designated HUL4 (YJR036C) and HUL5 (YGL141W), are viable. PMID- 9858557 TI - The repertoire of fos and jun proteins expressed during the G1 phase of the cell cycle is determined by the duration of mitogen-activated protein kinase activation. AB - In Rat-1 fibroblasts nonmitogenic doses of lysophosphatidic acid (LPA) stimulate a transient activation of mitogen-activated protein kinase (MAPK), whereas mitogenic doses elicit a sustained response. This sustained phase of MAPK activation regulates cell fate decisions such as proliferation or differentiation, presumably by inducing a program of gene expression which is not observed in response to transient MAPK activation. We have examined the expression of members of the AP-1 transcription factor complex in response to stimulation with different doses of LPA. c-Fos, c-Jun, and JunB are induced rapidly in response to LPA stimulation, whereas Fra-1 and Fra-2 are induced after a significant lag. The expression of c-Fos is transient, whereas the expression of c-Jun, JunB, Fra-1, and Fra-2 is sustained. The early expression of c-Fos can be reconstituted with nonmitogenic doses of LPA, but the response is transient compared to that observed with mitogenic doses. In contrast, expression of Fra-1, Fra-2, and JunB and optimal expression of c-Jun are observed only with doses of LPA which induce sustained MAPK activation and DNA synthesis. LPA-stimulated expression of c-Fos, Fra-1, Fra-2, c-Jun, and JunB is inhibited by the MEK1 inhibitor PD098059, indicating that the Raf-MEK-MAPK cascade is required for their expression. In cells expressing a conditionally active form of Raf-1 (DeltaRaf-1:ER), we observed that selective, sustained activation of Raf-MEK-MAPK was sufficient to induce expression of Fra-1, Fra-2, and JunB but, interestingly, induced little or no c-Fos or c-Jun. The induction of c-Fos observed in response to LPA was strongly inhibited by buffering the intracellular [Ca2+]. Moreover, although Raf activation or calcium ionophores induced little c-Fos expression, we observed a synergistic induction in response to the combination of DeltaRaf-1:ER and ionomycin. These results suggest that kinetically distinct phases of MAPK activation serve to regulate the expression of distinct AP-1 components such that sustained MAPK activation is required for the induced expression of Fra-1, Fra-2, c-Jun, and JunB. However, in contrast to the case for Fra-1, Fra-2, and JunB, activation of the MAPK cascade alone is not sufficient to induce c-Fos expression, which rather requires cooperation with other signals such as Ca2+ mobilization. Finally, the identification of the Fra-1, Fra-2, c-Jun, and JunB genes as genes which are selectively regulated by sustained MAPK activation or in response to activated Raf suggests that they are candidates to mediate certain of the effects of Ras proteins in oncogenic transformation. PMID- 9858559 TI - Cyclin D- and E-dependent kinases and the p57(KIP2) inhibitor: cooperative interactions in vivo. AB - This study examines in vivo the role and functional interrelationships of components regulating exit from the G1 resting phase into the DNA synthetic (S) phase of the cell cycle. Our approach made use of several key experimental attributes of the developing mouse lens, namely its strong dependence on pRb in maintenance of the postmitotic state, the down-regulation of cyclins D and E and up-regulation of the p57(KIP2) inhibitor in the postmitotic lens fiber cell compartment, and the ability to target transgene expression to this compartment. These attributes provide an ideal in vivo context in which to examine the consequences of forced cyclin expression and/or of loss of p57(KIP2) inhibitor function in a cellular compartment that permits an accurate quantitation of cellular proliferation and apoptosis rates in situ. Here, we demonstrate that, despite substantial overlap in cyclin transgene expression levels, D-type and E cyclins exhibited clear functional differences in promoting entry into S phase. In general, forced expression of the D-type cyclins was more efficient than cyclin E in driving lens fiber cells into S phase. In the case of cyclins D1 and D2, ectopic proliferation required their enhanced nuclear localization through CDK4 coexpression. High nuclear levels of cyclin E and CDK2, while not sufficient to promote efficient exit from G1, did act synergistically with ectopic cyclin D/CDK4. The functional differences between D-type and E cyclins was most evident in the p57(KIP2)-deficient lens wherein cyclin D overexpression induced a rate of proliferation equivalent to that of the pRb null lens, while overexpression of cyclin E did not increase the rate of proliferation over that induced by the loss of p57(KIP2) function. These in vivo analyses provide strong biological support for the prevailing view that the antecedent actions of cyclin D/CDK4 act cooperatively with cyclin E/CDK2 and antagonistically with p57(KIP2) to regulate the G1/S transition in a cell type highly dependent upon pRb. PMID- 9858560 TI - CA- and purine-rich elements form a novel bipartite exon enhancer which governs inclusion of the minute virus of mice NS2-specific exon in both singly and doubly spliced mRNAs. AB - The alternatively spliced 290-nucleotide NS2-specific exon of the parvovirus minute virus of mice (MVM), which is flanked by a large intron upstream and a small intron downstream, constitutively appears both in the R1 mRNA as part of a large 5'-terminal exon (where it is translated in open reading frame 3 [ORF3]), and in the R2 mRNA as an internal exon (where it is translated in ORF2). We have identified a novel bipartite exon enhancer element, composed of CA-rich and purine-rich elements within the 5' and 3' regions of the exon, respectively, that is required to include NS2-specific exon sequences in mature spliced mRNA in vivo. These two compositionally different enhancer elements are somewhat redundant in function: either element alone can at least partially support exon inclusion. They are also interchangeable: either element can function at either position. Either a strong 3' splice site upstream (i.e., the exon 5' terminus) or a strong 5' splice site downstream (i.e., the exon 3' terminus) is sufficient to prevent skipping of the NS2-specific exon, and a functional upstream 3' splice site is required for inclusion of the NS2-specific exon as an internal exon into the mature, doubly spliced R2 mRNA. The bipartite enhancer functionally strengthens these termini: the requirement for both the CA-rich and purine-rich elements can be overcome by improvements to the polypyrimidine tract of the upstream intron 3' splice site, and the purine-rich element also supports exon inclusion mediated through the downstream 5' splice sites. In summary, a suboptimal large-intron polypyrimidine tract, sequences within the downstream small intron, and a novel bipartite exonic enhancer operate together to yield the balanced levels of R1 and R2 observed in vivo. We suggest that the unusual bipartite exonic enhancer functions to mediate proper levels of inclusion of the NS2-specific exon in both singly spliced R1 and doubly spliced R2. PMID- 9858561 TI - SL1 trans splicing and 3'-end formation in a novel class of Caenorhabditis elegans operon. AB - Many Caenorhabditis elegans genes exist in operons in which polycistronic precursors are processed by cleavage at the 3' ends of upstream genes and trans splicing 100 to 400 nucleotides away, at the 5' ends of downstream genes, to generate monocistronic messages. Of the two spliced leaders, SL1 is trans spliced to the 5' ends of upstream genes, whereas SL2 is reserved for downstream genes in operons. However, there are isolated examples of what appears to be a different sort of operon, in which trans splicing is exclusively to SL1 and there is no intercistronic region; the polyadenylation signal is only a few base pairs upstream of the trans-splice site. We have analyzed the processing of an operon of this type by inserting the central part of mes-6/cks-1 into an SL2-type operon. In this novel context, cks-1 is trans spliced only to SL1, and mes-6 3' end formation occurs normally, demonstrating that this unique mode of processing is indeed intrinsic to this kind of operon, which we herein designate "SL1-type." An exceptionally long polypyrimidine tract found in the 3' untranslated regions of the three known SL1-type operons is shown to be required for the accumulation of both upstream and downstream mRNAs. Mutations of the trans-splice and poly(A) signals indicate that the two processes are independent and in competition, presumably due to their close proximity, raising the possibility that production of upstream and downstream mRNAs is mutually exclusive. PMID- 9858562 TI - Ribosomal protein L3 mutants alter translational fidelity and promote rapid loss of the yeast killer virus. AB - Programmed -1 ribosomal frameshifting is utilized by a number of RNA viruses as a means of ensuring the correct ratio of viral structural to enzymatic proteins available for viral particle assembly. Altering frameshifting efficiencies upsets this ratio, interfering with virus propagation. We have previously demonstrated that compounds that alter the kinetics of the peptidyl-transfer reaction affect programmed -1 ribosomal frameshift efficiencies and interfere with viral propagation in yeast. Here, the use of a genetic approach lends further support to the hypothesis that alterations affecting the ribosome's peptidyltransferase activity lead to changes in frameshifting efficiency and virus loss. Mutations in the RPL3 gene, which encodes a ribosomal protein located at the peptidyltransferase center, promote approximately three- to fourfold increases in programmed -1 ribosomal frameshift efficiencies and loss of the M1 killer virus of yeast. The mak8-1 allele of RPL3 contains two adjacent missense mutations which are predicted to structurally alter the Mak8-1p. Furthermore, a second allele that encodes the N-terminal 100 amino acids of L3 (called L3Delta) exerts a trans-dominant effect on programmed -1 ribosomal frameshifting and killer virus maintenance. Taken together, these results support the hypothesis that alterations in the peptidyltransferase center affect programmed -1 ribosomal frameshifting. PMID- 9858563 TI - The B29 (immunoglobulin beta-chain) gene is a genetic target for early B-cell factor. AB - Early B-cell factor (EBF) is a transcription factor suggested as essential for early B-lymphocyte development by findings in mice where the coding gene has been inactivated by homologous disruption. This makes the identification of genetic targets for this transcription factor pertinent for the understanding of early B cell development. The lack of B29 transcripts, coding for the beta subunit of the B-cell receptor complex, in pro-B cells from EBF-deficient mice suggested that B29 might be a genetic target for EBF. We here present data suggesting that EBF interacts with three independent sites within the mouse B29 promoter. Furthermore, ectopic expression of EBF in HeLa cells activated a B29 promoter controlled reporter construct 13-fold and induced a low level of expression from the endogenous B29 gene. Finally, mutations in the EBF binding sites diminished B29 promoter activity in pre-B cells while the same mutations did not have as striking an effect on the promoter function in B-cell lines of later differentiation stages. These data suggest that the B29 gene is a genetic target for EBF in early B-cell development. PMID- 9858565 TI - Testing for DNA tracking by MOT1, a SNF2/SWI2 protein family member. AB - Proteins in the SNF2/SWI2 family use ATP hydrolysis to catalyze rearrangements in diverse protein-DNA complexes. How ATP hydrolysis is coupled to these rearrangements is unknown, however. One attractive model is that these ATPases are ATP-dependent DNA-tracking enzymes. This idea was tested for the SNF2/SWI2 protein family member MOT1. MOT1 is an essential Saccharomyces cerevisiae transcription factor that uses ATP to dissociate TATA binding protein (TBP) from DNA. By using a series of DNA templates with one or two TATA boxes in combination with binding sites for heterologous DNA binding "roadblock" proteins, the ability of MOT1 to track along DNA was assayed. The results demonstrate that, following ATP-dependent TBP-DNA dissociation, MOT1 dissociates rapidly from the DNA by a mechanism that does not require a DNA end. Template commitment footprinting experiments support the conclusion that ATP-dependent DNA tracking by MOT1 does not occur. These results support a model in which MOT1 drives TBP-DNA dissociation by a mechanism that involves a transient, ATP-dependent interaction with TBP-DNA which does not involve ATP-dependent DNA tracking. PMID- 9858564 TI - A trans-activation domain in yeast heat shock transcription factor is essential for cell cycle progression during stress. AB - Gene expression in response to heat shock is mediated by the heat shock transcription factor (HSF), which in yeast harbors both amino- and carboxyl terminal transcriptional activation domains. Yeast cells bearing a truncated form of HSF in which the carboxyl-terminal transcriptional activation domain has been deleted [HSF(1-583)] are temperature sensitive for growth at 37 degreesC, demonstrating a requirement for this domain for sustained viability during thermal stress. Here we demonstrate that HSF(1-583) cells undergo reversible cell cycle arrest at 37 degreesC in the G2/M phase of the cell cycle and exhibit marked reduction in levels of the molecular chaperone Hsp90. As in higher eukaryotes, yeast possesses two nearly identical isoforms of Hsp90: one constitutively expressed and one highly heat inducible. When expressed at physiological levels in HSF(1-583) cells, the inducible Hsp90 isoform encoded by HSP82 more efficiently suppressed the temperature sensitivity of this strain than the constitutively expressed gene HSC82, suggesting that different functional roles may exist for these chaperones. Consistent with a defect in Hsp90 production, HSF(1-583) cells also exhibited hypersensitivity to the Hsp90-binding ansamycin antibiotic geldanamycin. Depletion of Hsp90 from yeast cells wild type for HSF results in cell cycle arrest in both G1/S and G2/M phases, suggesting a complex requirement for chaperone function in mitotic division during stress. PMID- 9858566 TI - FAST-2 is a mammalian winged-helix protein which mediates transforming growth factor beta signals. AB - The mechanisms by which transforming growth factor beta (TGF-beta) and related ligands regulate transcription remain poorly understood. The winged-helix (WH) transcription factor fork head activin signal transducer 1 (FAST-1) was identified as a mediator of activin signaling in Xenopus embryos (X. Chen, M. J. Rubock, and M. Whitman, Nature 383:691-696, 1996). We have cloned a novel WH gene from the mouse which shares many properties with FAST-1. We find that this gene, which we call FAST-2, is able to mediate transcriptional activation by TGF-beta. FAST-2 also interacts directly with Smad2, a cytoplasmic protein which is translocated to the nucleus in response to TGF-beta, and forms a multimeric complex with Smad2 and Smad4 on the activin response element, a high-affinity binding site for FAST-1. Analysis of the sequences of FAST-1 and FAST-2 reveals substantial protein sequence divergence compared to known vertebrate orthologs in the WH family. This suggests that FAST-2 represents a new WH gene related to FAST 1, which functions to mediate TGF-beta signals in mammals. We have also examined the structure of the FAST-2 gene and find that it overlaps with a kinesin motor protein gene. The genes are transcribed in opposite orientations, and their transcripts overlap in the 3' untranslated region. PMID- 9858567 TI - Two distinct gamma interferon-inducible promoters of the major histocompatibility complex class II transactivator gene are differentially regulated by STAT1, interferon regulatory factor 1, and transforming growth factor beta. AB - The major histocompatibility complex (MHC) class II transactivator (CIITA) is the master regulatory factor required for appropriate expression of class II MHC genes. Understanding the expression of CIITA is key to understanding the regulation of class II MHC genes. This report describes the independent regulation of two distinct CIITA promoters by cytokines with opposing functions, gamma interferon (IFN-gamma) and transforming growth factor beta (TGF-beta). A functional analysis of deletion mutations of the upstream promoter (promoter III) identified an IFN-gamma-responsive region located approximately 5 kb from the transcriptional start site. An in vivo DNase I hypersensitivity analysis detected a hypersensitive site in this area which supports the relevance of this region. When the downstream promoter (promoter IV) was studied by in vivo genomic footprinting, IFN-gamma-induced changes at putative binding sites for STAT1, interferon regulatory factor 1 (IRF-1), and E-box proteins were seen. Gel shift and supershift analyses for IRF-1 confirmed the in vivo footprint results. The role of the IFN-gamma-inducible transcription factor STAT1 was examined functionally. Although both promoters were controlled by STAT1, promoter-specific regulation was exhibited. The IFN-gamma response of promoter III was completely dependent on STAT1 and not IRF-1, while promoter IV was partially activated by IRF-1 in the total absence of STAT1 expression. While both promoters were affected by TGF-beta, activation of promoter III by IFN-gamma was more severely diminished by TGF-beta treatment. The differential control of CIITA promoters by TGF-beta, IRF-1, and STAT1 may be important in refining regulation of class II MHC genes in different cell types and under different stimulatory conditions. PMID- 9858568 TI - Receptor inhibition of pheromone signaling is mediated by the Ste4p Gbeta subunit. AB - The pheromone response pathway of the yeast Saccharomyces cerevisiae is initiated in MATa cells by binding of alpha-factor to the alpha-factor receptor. MATa cells in which the a-factor receptor is inappropriately expressed exhibit reduced pheromone signaling, a phenomenon termed receptor inhibition. In cells undergoing receptor inhibition, activation of the signaling pathway occurs normally at early time points but decreases after prolonged exposure to pheromone. Mutations that suppress the effects of receptor inhibition were obtained in the STE4 gene, which encodes the beta-subunit of the G protein that transmits the pheromone response signal. These mutations mapped to the N terminus and second WD repeat of Ste4p in regions that are not part of its Galpha binding surface. A STE4 allele containing several of these mutations, called STE4(SD13), reversed the signaling defect seen at late times in cells undergoing receptor inhibition but had no effect on the basal activity of the pathway. Moreover, the signaling properties of STE4(SD13) were indistinguishable from those of STE4 in wild-type MATa and MATalpha cells. These results demonstrate that the effect of the STE4(SD13) allele is specific to the receptor inhibition function of STE4. STE4(SD13) suppressed the signaling defect conferred by receptor inhibition in a MATa strain containing a deletion of GPA1, the G protein alpha-subunit gene; however, STE4(SD13) had no effect in a MATalpha strain containing a GPA1 deletion. Suppression of receptor inhibition by STE4(SD13) in a MATa strain containing a GPA1 deletion was unaffected by deletion of STE2, the alpha-factor receptor gene. The results presented here are consistent with a model in which an a-specific gene product other than Ste2p detects the presence of the a-factor receptor and blocks signaling by inhibiting the function of Ste4p. PMID- 9858569 TI - Cell differentiation during sexual development of the fungus Sordaria macrospora requires ATP citrate lyase activity. AB - During sexual development, mycelial cells from most filamentous fungi differentiate into typical fruiting bodies. Here, we describe the isolation and characterization of the Sordaria macrospora developmental mutant per5, which exhibits a sterile phenotype with defects in fruiting body maturation. Cytological investigations revealed that the mutant strain forms only ascus precursors without any mature spores. Using an indexed cosmid library, we were able to complement the mutant to fertility by DNA-mediated transformation. A single cosmid clone, carrying a 3.5-kb region able to complement the mutant phenotype, has been identified. Sequencing of the 3.5-kb region revealed an open reading frame of 2.1 kb interrupted by a 66-bp intron. The predicted polypeptide (674 amino acids) shows significant homology to eukaryotic ATP citrate lyases (ACLs), with 62 to 65% amino acid identity, and the gene was named acl1. The molecular mass of the S. macrospora ACL1 polypeptide is 73 kDa, as was verified by Western blot analysis with a hemagglutinin (HA) epitope-tagged ACL1 polypeptide. Immunological in situ detection of the HA-tagged polypeptide demonstrated that ACL is located within the cytosol. Sequencing of the mutant acl1 gene revealed a 1-nucleotide transition within the coding region, resulting in an amino acid substitution within the predicted polypeptide. Further evidence that ACL1 is essential for fruiting body maturation comes from experiments in which truncated and mutated versions of the acl1 gene were used for transformation. None of these copies was able to reconstitute the fertile phenotype in transformed per5 recipient strains. ACLs are usually involved in the formation of cytosolic acetyl coenzyme A (acetyl-CoA), which is used for the biosynthesis of fatty acids and sterols. Protein extracts from the mutant strain showed a drastic reduction in enzymatic activity compared to values obtained from the wild-type strain. Investigation of the time course of ACL expression suggests that ACL is specifically induced at the beginning of the sexual cycle and produces acetyl-CoA, which most probably is a prerequisite for fruiting body formation during later stages of sexual development. We discuss the contribution of ACL activity to the life cycle of S. macrospora. PMID- 9858570 TI - Functional role for protein kinase Cbeta as a regulator of stress-activated protein kinase activation and monocytic differentiation of myeloid leukemia cells. AB - Human myeloid leukemia cells respond to 12-O-tetradecanoylphorbol-13-acetate (TPA) and other activators of protein kinase C (PKC) with induction of monocytic differentiation. The present studies demonstrated that treatment of U-937 and HL 60 myeloid leukemia cells with TPA, phorbol-12,13-dibutyrate, or bryostatin 1 was associated with the induction of stress-activated protein kinase (SAPK). In contrast, TPA-resistant TUR and HL-525 cell variants deficient in PKCbeta failed to respond to activators of PKC with the induction of SAPK. A direct role for PKCbeta in TPA-induced SAPK activity in TUR and HL-525 cells that stably express PKCbeta was confirmed. We showed that TPA induced the association of PKCbeta with MEK kinase 1 (MEKK-1), an upstream effector of the SAPK/ERK kinase 1 (SEK1)- >SAPK cascade. The results also demonstrated that PKCbeta phosphorylated and activated MEKK-1 in vitro. The functional role of MEKK-1 in TPA-induced SAPK activity was further supported by the demonstration that the expression of a dominant negative MEKK-1 mutant abrogated this response. These findings indicate that PKCbeta activation is necessary for activation of the MEKK-1-->SEK1-->SAPK cascade in the TPA response of myeloid leukemia cells. PMID- 9858571 TI - The yeast RER2 gene, identified by endoplasmic reticulum protein localization mutations, encodes cis-prenyltransferase, a key enzyme in dolichol synthesis. AB - As an approach to understand the molecular mechanisms of endoplasmic reticulum (ER) protein sorting, we have isolated yeast rer mutants that mislocalize a Sec12 Mfalpha1p fusion protein from the ER to later compartments of the secretory pathway (S. Nishikawa and A. Nakano, Proc. Natl. Acad. Sci. USA 90:8179-8183, 1993). The temperature-sensitive rer2 mutant mislocalizes different types of ER membrane proteins, suggesting that RER2 is involved in correct localization of ER proteins in general. The rer2 mutant shows several other characteristic phenotypes: slow growth, defects in N and O glycosylation, sensitivity to hygromycin B, and abnormal accumulation of membranes, including the ER and the Golgi membranes. RER2 and SRT1, a gene whose overexpression suppresses rer2, encode novel proteins similar to each other, and their double disruption is lethal. RER2 homologues are found not only in eukaryotes but also in many prokaryote species and thus constitute a large gene family which has been well conserved during evolution. Taking a hint from the phenotype of newly established mutants of the Rer2p homologue of Escherichia coli, we discovered that the rer2 mutant is deficient in the activity of cis-prenyltransferase, a key enzyme of dolichol synthesis. This and other lines of evidence let us conclude that members of the RER2 family of genes encode cis-prenyltransferase itself. The difference in phenotypes between the rer2 mutant and previously obtained glycosylation mutants suggests a novel, as-yet-unknown role of dolichol. PMID- 9858572 TI - cDNA of the yeast retrotransposon Ty5 preferentially recombines with substrates in silent chromatin. AB - The yeast retrotransposon Ty5 preferentially integrates into regions of silent chromatin. Ty5 cDNA also recombines with homologous sequences, generating tandem elements or elements that have exchanged markers between cDNA and substrate. In this study, we demonstrate that Ty5 integration depends upon the conserved DD(35)E domain of integrase and cis-acting sequences at the end of the long terminal repeat (LTR) implicated in integrase binding. cDNA recombination requires Rad52p, which is responsible for homologous recombination. Interestingly, Ty5 cDNA recombines at least three times more frequently with substrates in silent chromatin than with a control substrate at an internal chromosomal locus. This preference depends upon the Ty5 targeting domain that is responsible for integration specificity, suggesting that localization of cDNA to silent chromatin results in the enhanced recombination. Recombination with a telomeric substrate occasionally generates highly reiterated Ty5 arrays, and mechanisms for tandem element formation were explored by using a plasmid-based recombination assay. Point mutations were introduced into plasmid targets, and recombination products were characterized to determine recombination initiation sites. Despite our previous observation of the importance of the LTR in forming tandem elements, recombination cannot simply be explained by crossover events between the LTRs of substrate and cDNA. We propose an alternative model based on single-strand annealing, where single-stranded cDNA initiates tandem element formation and the LTR is required for strand displacement to form a looped intermediate. Retrotransposons are increasingly found associated with chromosome ends, and amplification of Ty5 by both integration and recombination exemplifies how retroelements can contribute to telomere dynamics. PMID- 9858573 TI - CHOP-Dependent stress-inducible expression of a novel form of carbonic anhydrase VI. AB - CHOP (also called GADD153) is a stress-inducible nuclear protein that dimerizes with members of the C/EBP family of transcription factors and was initially identified as an inhibitor of C/EBP binding to classic C/EBP target genes. Subsequent experiments suggested a role for CHOP-C/EBP heterodimers in positively regulating gene expression; however, direct evidence that this is the case has so far not been uncovered. Here we describe the identification of a positively regulated direct CHOP-C/EBP target gene, that encoding murine carbonic anhydrase VI (CA-VI). The stress-inducible form of the gene is expressed from an internal promoter and encodes a novel intracellular form of what is normally a secreted protein. Stress-induced expression of CA-VI is both CHOP and C/EBPbeta dependent in that it does not occur in cells deficient in either gene. A CHOP-responsive element was mapped to the inducible CA-VI promoter, and in vitro footprinting revealed binding of CHOP-C/EBP heterodimers to that site. Rescue of CA-VI expression in c/ebpbeta-/- cells by exogenous C/EBPbeta and a shorter, normally inhibitory isoform of the protein known as LIP suggests that the role of the C/EBP partner is limited to targeting the CHOP-containing heterodimer to the response element and points to a preeminent role for CHOP in CA-VI induction during stress. PMID- 9858574 TI - A new 34-kilodalton isoform of human fibroblast growth factor 2 is cap dependently synthesized by using a non-AUG start codon and behaves as a survival factor. AB - Four isoforms of human fibroblast growth factor 2 (FGF-2) result from alternative initiations of translation at three CUG start codons and one AUG start codon. Here we characterize a new 34-kDa FGF-2 isoform whose expression is initiated at a fifth initiation codon. This 34-kDa FGF-2 was identified in HeLa cells by using an N-terminal directed antibody. Its initiation codon was identified by site directed mutagenesis as being a CUG codon located at 86 nucleotides (nt) from the FGF-2 mRNA 5' end. Both in vitro translation and COS-7 cell transfection using bicistronic RNAs demonstrated that the 34-kDa FGF-2 was exclusively expressed in a cap-dependent manner. This contrasted with the expression of the other FGF-2 isoforms of 18, 22, 22.5, and 24 kDa, which is controlled by an internal ribosome entry site (IRES). Strikingly, expression of the other FGF-2 isoforms became partly cap dependent in vitro in the presence of the 5,823-nt-long 3' untranslated region of FGF-2 mRNA. Thus, the FGF-2 mRNA can be translated both by cap-dependent and IRES-driven mechanisms, the balance between these two mechanisms modulating the ratio of the different FGF-2 isoforms. The function of the new FGF-2 was also investigated. We found that the 34-kDa FGF-2, in contrast to the other isoforms, permitted NIH 3T3 cell survival in low-serum conditions. A new arginine-rich nuclear localization sequence (NLS) in the N-terminal region of the 34-kDa FGF-2 was characterized and found to be similar to the NLS of human immunodeficiency virus type 1 Rev protein. These data suggest that the function of the 34-kDa FGF-2 is mediated by nuclear targets. PMID- 9858575 TI - Fiber-type-specific transcription of the troponin I slow gene is regulated by multiple elements. AB - The regulatory elements that restrict transcription of genes encoding contractile proteins specifically to either slow- or fast-twitch skeletal muscles are unknown. As an initial step towards understanding the mechanisms that generate muscle diversity during development, we have identified a 128-bp troponin I slow upstream element (SURE) and a 144-bp troponin I fast intronic element (FIRE) that confer fiber type specificity in transgenic mice (M. Nakayama et al., Mol. Cell. Biol. 16:2408-2417, 1996). SURE and FIRE have maintained the spatial organization of four conserved motifs (3' to 5'): an E box, an AT-rich site (A/T2) that binds MEF-2, a CACC site, and a novel CAGG motif. Troponin I slow (TnIs) constructs harboring mutations in these motifs were analyzed in transiently and stably transfected Sol8 myocytes and in transgenic mice to assess their function. Mutations of the E-box, A/T2, and CAGG motifs completely abolish transcription from the TnI SURE. In contrast, mutation of the CACC motif had no significant effect in transfected myocytes or on the slow-specific transcription of the TnI SURE in transgenic mice. To assess the role of E boxes in fiber type specificity, a chimeric enhancer was constructed in which the E box of SURE was replaced with the E box from FIRE. This TnI E box chimera, which lacks the SURE NFAT site, confers essentially the same levels of transcription in transgenic mice as those conferred by wild-type SURE and is specifically expressed in slow-twitch muscles, indicating that the E box on its own cannot determine the fiber-type-specific expression of the TnI promoter. The importance of the 5' half of SURE, which bears little homology to the TnI FIRE, in muscle-specific expression was analyzed by deletion and linker scanning analyses. Removal of the 5' half of SURE (-846 to -811) results in the loss of expression in stably transfected but not in transiently expressing myocytes. Linker scanning mutations identified sequences in this region that are necessary for the function of SURE when integrated into chromatin. One of these sites (GTTAATCCG), which is highly homologous to a bicoid consensus site, binds to nuclear proteins from several mesodermal cells. These results show that multiple elements are involved in the muscle-specific activity of the TnIs promoter and that interactions between upstream and downstream regions of SURE are important for transcription in the context of native chromatin. PMID- 9858576 TI - Identification of the in vivo casein kinase II phosphorylation site within the homeodomain of the cardiac tisue-specifying homeobox gene product Csx/Nkx2.5. AB - Csx/Nkx2.5, a member of the homeodomain-containing transcription factors, serves critical developmental functions in heart formation in vertebrates and nonvertebrates. In this study the putative nuclear localization signal (NLS) of Csx/Nkx2.5 was identified by site-directed mutagenesis to the amino terminus of the homeodomain, which is conserved in almost all homeodomain proteins. When the putative NLS of Csx/Nkx2.5 was mutated a significant amount of the cytoplasmically localized Csx/Nkx2.5 was unphosphorylated, in contrast to the nuclearly localized Csx/Nkx2.5, which is serine- and threonine-phosphorylated, suggesting that Csx/Nkx2.5 phosphorylation is regulated, at least in part, by intracellular localization. Tryptic phosphopeptide mapping indicated that Csx/Nkx2.5 has at least five phosphorylation sites. Using in-gel kinase assays, we detected a Csx/Nkx2.5 kinase whose molecular mass is approximately 40 kDa in both cytoplasmic and nuclear extracts. Mutational analysis and in vitro kinase assays suggested that this 40-kDa Csx/Nkx2.5 kinase is a catalytic subunit of casein kinase II (CKII) that phosphorylates the serine residue between the first and second helix of the homeodomain. This CKII site is phosphorylated in vivo. CKII-dependent phosphorylation of the homeodomain increased Csx/Nkx2. 5 DNA binding. Serine-to-alanine mutation at the CKII phosphorylation site reduced transcriptional activity when the carboxyl-terminal repressor domain was deleted. Although the precise biological function of Csx/Nkx2.5 phosphorylation by CKII remains to be determined, it may play an important role, as this CKII phosphorylation site within the homeodomain is fully conserved in all known members of the NK2 family of the homeobox genes. PMID- 9858577 TI - Repressors and upstream repressing sequences of the stress-regulated ENA1 gene in Saccharomyces cerevisiae: bZIP protein Sko1p confers HOG-dependent osmotic regulation. AB - The yeast ENA1/PMR2A gene encodes a cation extrusion ATPase in Saccharomyces cerevisiae which is essential for survival under salt stress conditions. One important mechanism of ENA1 transcriptional regulation is based on repression under normal growth conditions, which is relieved by either osmotic induction or glucose starvation. Analysis of the ENA1 promoter revealed a Mig1p-binding motif (-533 to -544) which was characterized as an upstream repressing sequence (URSMIG ENA1) regulated by carbon source. Its function was abolished in a mig1 mig2 double-deletion strain as well as in either ssn6 or tup1 single mutants. A second URS at -502 to -513 is responsible for transcriptional repression regulated by osmotic stress and is similar to mammalian cyclic AMP response elements (CREs) that are recognized by CREB proteins. This URSCRE-ENA1 element requires for its repression function the yeast CREB homolog Sko1p (Acr1p) as well as the integrity of the Ssn6p-Tup1p corepressor complex. When targeted to the GAL1 promoter by fusing with the Gal4p DNA-binding domain, Sko1p acts as an Ssn6/Tup1p-dependent repressor regulated by osmotic stress. A glutathione S-transferase-Sko1 fusion protein binds specifically to the URSCRE-ENA1 element. Furthermore, a hog1 mitogen-activated protein kinase deletion strain could not counteract repression on URSCRE-ENA1 during osmotic shock. The loss of SKO1 completely restored ENA1 expression in a hog1 mutant and partially suppressed the osmotic stress sensitivity, qualifying Sko1p as a downstream effector of the HOG pathway. Our results indicate that different signalling pathways (HOG osmotic pathway and glucose repression pathway) use distinct promoter elements of ENA1 (URSCRE-ENA1 and URSMIG-ENA1) via specific transcriptional repressors (Sko1p and Mig1/2p) and via the general Ssn6p-Tup1p complex. The physiological importance of the relief from repression during salt stress was also demonstrated by the increased tolerance of sko1 or ssn6 mutants to Na+ or Li+ stress. PMID- 9858578 TI - Specification of regions of DNA replication initiation during embryogenesis in the 65-kilobase DNApolalpha-dE2F locus of Drosophila melanogaster. AB - In the early stage of Drosophila embryogenesis, DNA replication initiates at unspecified sites in the chromosome. In contrast, DNA replication initiates in specified regions in cultured cells. We investigated when and where the initiation regions are specified during embryogenesis and compared them with those observed in cultured cells by two-dimensional gel methods. In the DNA polymerase alpha gene (DNApolalpha) locus, where an initiation region, oriDalpha, had been identified in cultured Kc cells, repression of origin activity in the coding region was detected after formation of cellular blastoderms, and the range of the initiation region had become confined by 5 h after fertilization. During this work we identified other initiation regions between oriDalpha and the Drosophila E2F gene (dE2F) downstream of DNApolalpha. At least four initiation regions showing replication bubbles were identified in the 65-kb DNApolalpha-dE2F locus in 5-h embryos, but only two were observed in Kc cells. These results suggest that the specification levels of origin usage in 5-h embryos are in the intermediate state compared to those in more differentiated cells. Further, we found a spatial correlation between the active promoter regions for dE2F and the active initiation zones of replication. In 5-h embryos, two known transcripts differing in their first exons were expressed, and two regions close to the respective promoter regions for both transcripts functioned as replication origins. In Kc cells, only one transcript was expressed and functional replication origins were observed only in the region including the promoter region for this transcript. PMID- 9858579 TI - The nuclease activity of Mre11 is required for meiosis but not for mating type switching, end joining, or telomere maintenance. AB - The Saccharomyces cerevisiae MRE11 gene is required for the repair of ionizing radiation-induced DNA damage and for the initiation of meiotic recombination. Sequence analysis has revealed homology between Mre11 and SbcD, the catalytic subunit of an Escherichia coli enzyme with endo- and exonuclease activity, SbcCD. In this study, the purified Mre11 protein was found to have single-stranded endonuclease activity. This activity was absent from mutant proteins containing single amino acid substitutions in either one of two sequence motifs that are shared by Mre11 and SbcD. Mutants with allele mre11-D56N or mre11-H125N were partially sensitive to ionizing radiation but lacked the other mitotic phenotypes of poor vegetative growth, hyperrecombination, defective nonhomologous end joining, and shortened telomeres that are characteristic of the mre11 null mutant. Diploids homozygous for the mre11-H125N mutation failed to sporulate and accumulated unresected double-strand breaks (DSB) during meiosis. We propose that in mitotic cells DSBs can be processed by other nucleases that are partially redundant with Mre11, but these activities are unable to process Spo11-bound DSBs in meiotic cells. PMID- 9858580 TI - A box H/ACA small nucleolar RNA-like domain at the human telomerase RNA 3' end. AB - Simple sequence repeat telomeric DNA is maintained by a specialized reverse transcriptase, telomerase. The integral RNA subunit of telomerase contains a template region that determines the sequence added to chromosome ends. Aside from providing the template, little is known about the role of the telomerase RNA. In addition, no hypotheses have been suggested to account for the striking evolutionary divergence in size and sequence between telomerase RNAs of ciliates, yeasts, and mammals. We show that the two- to threefold increase in size of the mammalian telomerase RNAs relative to ciliate telomerase RNAs is due to the presence of an extra domain resembling a box H/ACA small nucleolar RNA (snoRNA). The human telomerase RNA (hTR) H/ACA domain is essential in vivo for hTR accumulation, hTR 3' end processing, and telomerase activity. By substituting the U64 box H/ACA snoRNA for the hTR H/ACA domain, we demonstrate that a heterologous snoRNA can function to promote chimeric RNA accumulation and 3' end processing but not telomerase activity. In addition, we show that maturation of full-length hTR and its assembly into active telomerase occur from an mRNA promoter-driven RNA polymerase II transcript but not from a U6 snRNA promoter-driven RNA polymerase III transcript. Finally, we show that a small percentage of hTR is associated with nucleoli. These results have implications for the biogenesis and structure of hTR and the human telomerase ribonucleoprotein complex. They also expand the structural and functional diversity of the box H/ACA snoRNA motif. PMID- 9858581 TI - Elevated levels of a U4/U6.U5 snRNP-associated protein, Spp381p, rescue a mutant defective in spliceosome maturation. AB - U4 snRNA release from the spliceosome occurs through an essential but ill-defined Prp38p-dependent step. Here we report the results of a dosage suppressor screen to identify genes that contribute to PRP38 function. Elevated expression of a previously uncharacterized gene, SPP381, efficiently suppresses the growth and splicing defects of a temperature-sensitive (Ts) mutant prp38-1. This suppression is specific in that enhanced SPP381 expression does not alter the abundance of intronless RNA transcripts or suppress the Ts phenotypes of other prp mutants. Since SPP381 does not suppress a prp38::LEU2 null allele, it is clear that Spp381p assists Prp38p in splicing but does not substitute for it. Yeast SPP381 disruptants are severely growth impaired and accumulate unspliced pre-mRNA. Immune precipitation studies show that, like Prp38p, Spp381p is present in the U4/U6.U5 tri-snRNP particle. Two-hybrid analyses support the view that the carboxyl half of Spp381p directly interacts with the Prp38p protein. A putative PEST proteolysis domain within Spp381p is dispensable for the Spp381p-Prp38p interaction and for prp38-1 suppression but contributes to Spp381p function in splicing. Curiously, in vitro, Spp381p may not be needed for the chemistry of pre mRNA splicing. Based on the in vivo and in vitro results presented here, we propose that two small acidic proteins without obvious RNA binding domains, Spp381p and Prp38p, act in concert to promote U4/U5.U6 tri-snRNP function in the spliceosome cycle. PMID- 9858582 TI - The yeast a1 and alpha2 homeodomain proteins do not contribute equally to heterodimeric DNA binding. AB - In diploid cells of the yeast Saccharomyces cerevisiae, the alpha2 and a1 homeodomain proteins bind cooperatively to sites in the promoters of haploid cell type-specific genes (hsg) to repress their expression. Although both proteins bind to the DNA, in the alpha2 homeodomain substitutions of residues that are involved in contacting the DNA have little or no effect on repression in vivo or cooperative DNA binding with a1 protein in vitro. This result brings up the question of the contribution of each protein in the heterodimer complex to the DNA-binding affinity and specificity. To determine the requirements for the a1 alpha2 homeodomain DNA recognition, we systematically introduced single base-pair substitutions in an a1-alpha2 DNA-binding site and examined their effects on repression in vivo and DNA binding in vitro. Our results show that nearly all substitutions that significantly decrease repression and DNA-binding affinity are at positions which are specifically contacted by either the alpha2 or a1 protein. Interestingly, an alpha2 mutant lacking side chains that make base-specific contacts in the major groove is able to discriminate between the wild-type and mutant DNA sites with the same sequence specificity as the wild-type protein. These results suggest that the specificity of alpha2 DNA binding in complex with a1 does not rely solely on the residues that make base-specific contacts. We have also examined the contribution of the a1 homeodomain to the binding affinity and specificity of the complex. In contrast to the lack of a defective phenotype produced by mutations in the alpha2 homeodomain, many of the alanine substitutions of residues in the a1 homeodomain have large effects on a1-alpha2 mediated repression and DNA binding. This result shows that the two proteins do not make equal contributions to the DNA-binding affinity of the complex. PMID- 9858583 TI - The oncogenic potential of the Pax3-FKHR fusion protein requires the Pax3 homeodomain recognition helix but not the Pax3 paired-box DNA binding domain. AB - The chimeric transcription factor Pax3-FKHR, produced by the t(2;13)(q35;q14) chromosomal translocation in alveolar rhabdomyosarcoma, consists of the two Pax3 DNA binding domains (paired box and homeodomain) fused to the C-terminal forkhead (FKHR) sequences that contain a potent transcriptional activation domain. To determine which of these domains are required for cellular transformation, Pax3, Pax3-FKHR, and selected mutants were retrovirally expressed in NIH 3T3 cells and evaluated for their capacity to promote anchorage-independent cell growth. Mutational analysis revealed that both the third alpha-helix of the homeodomain and a small region of the FKHR transactivation domain are absolutely required for efficient transformation by the Pax3-FKHR fusion protein. Surprisingly, point mutations in the paired domain that abrogate sequence-specific DNA binding retained transformation potential equivalent to that of the wild-type protein. This finding suggests that DNA binding mediated through the Pax3 paired box is not required for transformation. Our results demonstrate that the integrity of the Pax3 homeodomain recognition helix and the FKHR transactivation domain is necessary for efficient cellular transformation by the Pax3-FKHR fusion protein. PMID- 9858584 TI - Genetic evidence for Pak1 autoinhibition and its release by Cdc42. AB - Pak1 protein kinase of Schizosaccharomyces pombe, a member of the p21-GTPase activated protein kinase (PAK) family, participates in signaling pathways including sexual differentiation and morphogenesis. The regulatory domain of PAK proteins is thought to inhibit the kinase catalytic domain, as truncation of this region renders kinases more active. Here we report the detection in the two hybrid system of the interaction between Pak1 regulatory domain and the kinase catalytic domain. Pak1 catalytic domain binds to the same highly conserved region on the regulatory domain that binds Cdc42, a GTPase protein capable of activating Pak1. Two-hybrid, mutant, and genetic analyses indicated that this intramolecular interaction rendered the kinase in a closed and inactive configuration. We show that Cdc42 can induce an open configuration of Pak1. We propose that Cdc42 interaction disrupts the intramolecular interactions of Pak1, thereby releasing the kinase from autoinhibition. PMID- 9858585 TI - Cyclin E2, a novel G1 cyclin that binds Cdk2 and is aberrantly expressed in human cancers. AB - A novel cyclin gene was discovered by searching an expressed sequence tag database with a cyclin box profile. The human cyclin E2 gene encodes a 404-amino acid protein that is most closely related to cyclin E. Cyclin E2 associates with Cdk2 in a functional kinase complex that is inhibited by both p27(Kip1) and p21(Cip1). The catalytic activity associated with cyclin E2 complexes is cell cycle regulated and peaks at the G1/S transition. Overexpression of cyclin E2 in mammalian cells accelerates G1, demonstrating that cyclin E2 may be rate limiting for G1 progression. Unlike cyclin E1, which is expressed in most proliferating normal and tumor cells, cyclin E2 levels were low to undetectable in nontransformed cells and increased significantly in tumor-derived cells. The discovery of a novel second cyclin E family member suggests that multiple unique cyclin E-CDK complexes regulate cell cycle progression. PMID- 9858586 TI - Cloning, characterization, and expression of a novel Zn2+-binding FYVE finger containing phosphoinositide kinase in insulin-sensitive cells. AB - Signaling by phosphorylated species of phosphatidylinositol (PI) appears to regulate diverse responses in eukaryotic cells. A differential display screen for fat- and muscle-specific transcripts led to identification and cloning of the full-length cDNA of a novel mammalian 2,052-amino-acid protein (p235) from a mouse adipocyte cDNA library. Analysis of the deduced amino acid sequence revealed that p235 contains an N-terminal zinc-binding FYVE finger, a chaperonin like region in the middle of the molecule, and a consensus for phosphoinositide 5 kinases at the C terminus. p235 mRNA appears as a 9-kb transcript, enriched in insulin-sensitive cells and tissues, likely transcribed from a single-copy gene in at least two close-in-size splice variants. Specific antibodies against mouse p235 were raised, and both the endogenously and heterologously expressed proteins were biochemically detected in 3T3-L1 adipocytes and transfected COS cells, respectively. Immunofluorescence microscopy analysis of endogenous p235 localization in 3T3-L1 adipocytes with affinity-purified anti-p235 antibodies documented a punctate peripheral pattern. In COS cells, the expressed p235 N terminal but not the C-terminal region displayed a vesicular pattern similar to that in 3T3-L1 adipocytes that became diffuse upon Zn2+ chelation or FYVE finger truncation. A recombinant protein comprising the N-terminal but not the C terminal region of the molecule was found to bind 2.2 mole equivalents of Zn2+. Determination of the lipid kinase activity in the p235 immunoprecipitates derived from 3T3-L1 adipocytes or from COS cells transiently expressing p235 revealed that p235 displayed unique preferences for PI substrate over already phosphorylated PI. In conclusion, the mouse p235 protein determines an important novel class of phosphoinositide kinases that seems to be targeted to specific intracellular loci by a Zn-dependent mechanism. PMID- 9858587 TI - Regulation of cyclin A-Cdk2 by SCF component Skp1 and F-box protein Skp2. AB - Cyclin A-Cdk2 complexes bind to Skp1 and Skp2 during S phase, but the function of Skp1 and Skp2 is unclear. Skp1, together with F-box proteins like Skp2, are part of ubiquitin-ligase E3 complexes that target many cell cycle regulators for ubiquitination-mediated proteolysis. In this study, we investigated the potential regulation of cyclin A-Cdk2 activity by Skp1 and Skp2. We found that Skp2 can inhibit the kinase activity of cyclin A-Cdk2 in vitro, both by direct inhibition of cyclin A-Cdk2 and by inhibition of the activation of Cdk2 by cyclin-dependent kinase (CDK)-activating kinase phosphorylation. Only the kinase activity of Cdk2, not of that of Cdc2 or Cdk5, is reduced by Skp2. Skp2 is phosphorylated by cyclin A-Cdk2 on residue Ser76, but nonphosphorylatable mutants of Skp2 can still inhibit the kinase activity of cyclin A-Cdk2 toward histone H1. The F box of Skp2 is required for binding to Skp1, and both the N-terminal and C-terminal regions of Skp2 are involved in binding to cyclin A-Cdk2. Furthermore, Skp2 and the CDK inhibitor p21(Cip1/WAF1) bind to cyclin A-Cdk2 in a mutually exclusive manner. Overexpression of Skp2, but not Skp1, in mammalian cells causes a G1/S cell cycle arrest. PMID- 9858588 TI - Cell cycle-dependent regulation of human DNA polymerase alpha-primase activity by phosphorylation. AB - DNA polymerase alpha-primase is known to be phosphorylated in human and yeast cells in a cell cycle-dependent manner on the p180 and p68 subunits. Here we show that phosphorylation of purified human DNA polymerase alpha-primase by purified cyclin A/cdk2 in vitro reduced its ability to initiate simian virus 40 (SV40) DNA replication in vitro, while phosphorylation by cyclin E/cdk2 stimulated its initiation activity. Tryptic phosphopeptide mapping revealed a family of p68 peptides that was modified well by cyclin A/cdk2 and poorly by cyclin E/cdk2. The p180 phosphopeptides were identical with both kinases. By mass spectrometry, the p68 peptide family was identified as residues 141 to 160. Cyclin A/cdk2- and cyclin A/cdc2-modified p68 also displayed a phosphorylation-dependent shift to slower electrophoretic mobility. Mutation of the four putative phosphorylation sites within p68 peptide residues 141 to 160 prevented its phosphorylation by cyclin A/cdk2 and the inhibition of replication activity. Phosphopeptide maps of the p68 subunit of DNA polymerase alpha-primase from human cells, synchronized and labeled in G1/S and in G2, revealed a cyclin E/cdk2-like pattern in G1/S and a cyclin A/cdk2-like pattern in G2. The slower-electrophoretic-mobility form of p68 was absent in human cells in G1/S and appeared as the cells entered G2/M. Consistent with this, the ability of DNA polymerase alpha-primase isolated from synchronized human cells to initiate SV40 replication was maximal in G1/S, decreased as the cells completed S phase, and reached a minimum in G2/M. These results suggest that the replication activity of DNA polymerase alpha-primase in human cells is regulated by phosphorylation in a cell cycle-dependent manner. PMID- 9858589 TI - Direct regulation of mitochondrial RNA synthesis by thyroid hormone. AB - We have analyzed the influence of in vivo treatment and in vitro addition of thyroid hormone on in organello mitochondrial DNA (mtDNA) transcription and, in parallel, on the in organello footprinting patterns at the mtDNA regions involved in the regulation of transcription. We found that thyroid hormone modulates mitochondrial RNA levels and the mRNA/rRNA ratio by influencing the transcriptional rate. In addition, we found conspicuous differences between the mtDNA dimethyl sulfate footprinting patterns of mitochondria derived from euthyroid and hypothyroid rats at the transcription initiation sites but not at the mitochondrial transcription termination factor (mTERF) binding region. Furthermore, direct addition of thyroid hormone to the incubation medium of mitochondria isolated from hypothyroid rats restored the mRNA/rRNA ratio found in euthyroid rats as well as the mtDNA footprinting patterns at the transcription initiation area. Therefore, we conclude that the regulatory effect of thyroid hormone on mitochondrial transcription is partially exerted by a direct influence of the hormone on the mitochondrial transcription machinery. Particularly, the influence on the mRNA/rRNA ratio is achieved by selective modulation of the alternative H-strand transcription initiation sites and does not require the previous activation of nuclear genes. These results provide the first functional demonstration that regulatory signals, such as thyroid hormone, that modify the expression of nuclear genes can also act as primary signals for the transcriptional apparatus of mitochondria. PMID- 9858590 TI - Analysis of mice with single and multiple copies of transgenes reveals a novel arrangement for the lambda5-VpreB1 locus control region. AB - The murine lambda5-VpreB1 locus encodes two proteins that form part of the pre-B cell receptor and play a key role in B-lymphocyte development. We have identified a locus control region (LCR) which is responsible for coordinate activation of both genes in pre-B cells. Analysis of mice with single and multiple copies of transgenes shows a clear difference in the expression behavior of the genes depending on the transgene copy number. While expression of both lambda5 and VpreB1 in single- and two-copy integrations requires the presence of a set of DNase I hypersensitive sites located 3' of the lambda5 gene, small fragments containing the genes have LCR activity when arranged in multiple-copy tandem arrays, indicating that additional components of the LCR are located within or close to the genes. The complete LCR is capable of driving efficient copy dependent expression of a lambda5 gene in pre-B cells even when it is integrated into centomeric gamma-satellite DNA. The finding that activation of expression of the locus by positively acting factors is fully dominant over the silencing effect of heterochromatin has implications for models for chromatin-mediated gene silencing during B-cell development. PMID- 9858592 TI - Characterization of the DNA-binding and dimerization properties of the nuclear orphan receptor germ cell nuclear factor. AB - The orphan receptor germ cell nuclear factor (GCNF) is a member of the superfamily of nuclear receptors. During development, GCNF exhibits a restricted brain-specific expression pattern, whereas GCNF expression in the adult is germ cell specific. Therefore, the receptor may participate in the regulation of neurogenesis and reproductive functions. No natural GCNF target gene has yet been identified, but recent data demonstrate specific and high-affinity binding of GCNF either to the direct repeat DNA element AGGTCAAGGTCA (DR0) or to extended half-sites, such as TCAAGGTCA. In this study, we show that murine GCNF (mGCNF) can bind as a homodimer to extended half-sites, thus describing a novel property within the nuclear receptor superfamily. Homodimeric binding to extended half sites requires the presence of a dimerization function within the mGCNF DNA binding domain (DBD) and a novel dimerization surface encompassing the putative helix 3 and the helix 12 region of the mGCNF ligand-binding domain (LBD). In addition, the mGCNF LBD has the potential to adopt different conformations with distinct dimerization properties. The helix 12 region of the mGCNF LBD not only regulates the switch between these dimerization conformations but also dictates the DNA-binding behavior and transcriptional properties of the different dimerization conformations. In summary, our findings describe unique DNA-binding and dimerization properties of a nuclear receptor and suggest a novel mechanism that allows mGCNF to modulate target gene activity. PMID- 9858591 TI - trans repression of the human metallothionein IIA gene promoter by PZ120, a novel 120-kilodalton zinc finger protein. AB - Metallothioneins are small, highly conserved, cysteine-rich proteins that bind a variety of metal ions. They are found in virtually all eukaryotic organisms and are regulated primarily at the transcriptional level. In humans, the predominant metallothionein gene is hMTIIA, which accounts for 50% of all metallothioneins expressed in cultured human cells. The hMTIIA promoter is quite complex. In addition to cis-acting DNA sequences that serve as binding sites for trans-acting factors such as Sp1, AP1, AP2, AP4, and the glucocorticoid receptor, the hMTIIA promoter contains eight consensus metal response element sequences. We report here the cloning of a novel zinc finger protein with a molecular mass of 120 kDa (PZ120) that interacts specifically with the hMTIIA transcription initiation site. The PZ120 protein is ubiquitously expressed in most tissues and possesses a conserved poxvirus and zinc finger (POZ) motif previously found in several zinc finger transcription factors. Intriguingly, we found that a region of PZ120 outside of the zinc finger domain can bind specifically to the hMTIIA DNA. Using transient-transfection analysis, we found that PZ120 repressed transcription of the hMTIIA promoter. These results suggest that the hMTIIA gene is regulated by an additional negative regulator that has not been previously described. PMID- 9858593 TI - The NeuroD1/BETA2 sequences essential for insulin gene transcription colocalize with those necessary for neurogenesis and p300/CREB binding protein binding. AB - NeuroD1/BETA2 is a key regulator of pancreatic islet morphogenesis and insulin hormone gene transcription in islet beta cells. This factor also appears to be involved in neurogenic differentiation, because NeuroD1/BETA2 is able to induce premature differentiation of neuronal precursors and convert ectoderm into fully differentiated neurons upon ectopic expression in Xenopus embryos. We have identified amino acid sequences in mammalian and Xenopus NeuroD1/BETA2 that are necessary for insulin gene expression and ectopic neurogenesis. Our results indicate that evolutionarily conserved sequences spanning the basic helix-loop helix (amino acids [aa] 100 to 155) and C-terminal (aa 156 to 355) regions are important for both of these processes. The transactivation domains (AD1, aa 189 to 299; AD2, aa 300 to 355) were within the carboxy-terminal region, as analyzed by using GAL4:NeuroD1/BETA2 chimeras. Selective activation of mammalian insulin gene enhancer-driven expression and ectopic neurogenesis in Xenopus embryos was regulated by two independent and separable domains of NeuroD1/BETA2, located between aa 156 to 251 and aa 252 to 355. GAL4:NeuroD1/BETA2 constructs spanning these sequences demonstrated that only aa 252 to 355 contained activation domain function, although both aa 156 to 251 and 300 to 355 were found to interact with the p300/CREB binding protein (CBP) coactivator. These results implicate p300/CBP in NeuroD1/BETA2 function and further suggest that comparable mechanisms are utilized to direct target gene transcription during differentiation and in adult islet beta cells. PMID- 9858594 TI - RGS3 inhibits G protein-mediated signaling via translocation to the membrane and binding to Galpha11. AB - In the present study, we investigated the function and the mechanism of action of RGS3, a member of a family of proteins called regulators of G protein signaling (RGS). Polyclonal antibodies against RGS3 were produced and characterized. An 80 kDa protein was identified as RGS3 by immunoprecipitation and immunoblotting with anti-RGS3 antibodies in a human mesangial cell line (HMC) stably transfected with RGS3 cDNA. Coimmunoprecipitation experiments in RGS3-overexpressing cell lysates revealed that RGS3 bound to aluminum fluoride-activated Galpha11 and to a lesser extent to Galphai3 and that this binding was mediated by the RGS domain of RGS3. A role of RGS3 in postreceptor signaling was demonstrated by decreased calcium responses and mitogen-activated protein (MAP) kinase activity induced by endothelin-1 in HMC stably overexpressing RGS3. Moreover, depletion of endogenous RGS3 by transfection of antisense RGS3 cDNA in NIH 3T3 cells resulted in enhanced MAP kinase activation induced by endothelin-1. The study of intracellular distribution of RGS3 indicated its unique cytosolic localization. Activation of G proteins by AlF4-, NaF, or endothelin-1 resulted in redistribution of RGS3 from cytosol to the plasma membrane as determined by Western blotting of the cytosolic and particulate fractions with RGS3 antiserum as well as by immunofluorescence microscopy. Agonist-induced translocation of RGS3 occurred by a dual mechanism involving both C-terminal (RGS domain) and N-terminal regions of RGS3. Thus, coexpression of RGS3 with a constitutively active mutant of Galpha11 (Galpha11 QL) resulted in the binding of RGS3, but not of its N-terminal fragment, to the membrane fraction and in its interaction with Galpha11-QL in vitro without any stimuli. However, both full-length RGS3 and its N-terminal domain translocated to the plasma membrane upon stimulation of intact cells with endothelin-1 as assayed by immunofluorescence microscopy. The effect of endothelin-1 was also mimicked by calcium ionophore A23187, suggesting the importance of Ca2+ in the mechanism of redistribution of RGS3. These data indicate that RGS3 inhibits G protein-coupled receptor signaling by a complex mechanism involving its translocation to the membrane in addition to its established function as a GTPase-activating protein. PMID- 9858595 TI - Siah-1 N-terminal RING domain is required for proteolysis function, and C terminal sequences regulate oligomerization and binding to target proteins. AB - The Drosophila seven in absentia (sina) gene was initially discovered because its inactivation leads to R7 photoreceptor defects. Recent data indicate that Sina binds to the Sevenless pathway protein Phyllopod, and together they mediate degradation of Tramtrack, a transcriptional repressor of R7 cell fate. Independent studies have shown that Sina and its highly related mammalian homologues Siah-1 and Siah-2 bind to the DCC (deleted in colorectal cancer) protein and promote its proteolysis via the ubiquitin-proteasome pathway. To determine the roles of mammalian Siahs in proteolysis and their interactions with target proteins, we sought to define Siah-1 domains critical for regulation of DCC. Mutant Siah-1 proteins, harboring missense mutations in the carboxy (C) terminal domain analogous to those present in Drosophila sina loss-of-function alleles, failed to promote DCC degradation. Point mutations and deletion of the amino (N)-terminal RING finger domain of Siah-1 abrogated its ability to promote DCC proteolysis. In the course of defining Siah-1 sequences required for DCC degradation, we found that Siah-1 is itself rapidly degraded via the proteasome pathway, and RING domain mutations stabilized the Siah-1 protein. Siah-1 was found to oligomerize with itself and other Sina and Siah proteins via C-terminal sequences. Finally, evidence that endogenous Siah-1 regulates DCC proteolysis in cells was obtained through studies of an apparent dominant negative mutant of Siah-1, as well as via an antisense approach. The data indicate that the Siah-1 N terminal RING domain is required for its proteolysis function, while the C terminal sequences regulate oligomerization and binding to target proteins, such as DCC. PMID- 9858597 TI - Melanization of Cryptococcus neoformans in murine infection. AB - Cryptococcus neoformans is a fungus that is pathogenic in humans and that can produce melanin in vitro. Melanization is associated with virulence, but there is no evidence that melanin is made during infection. Melanins are difficult to study because they are amorphous and insoluble. Melanin-binding peptides from a phage display library were used to demonstrate that C. neoformans makes melanin like compounds in tissue. Melanin-binding peptides were characterized by a high proportion of positively charged and aromatic residues. Two other methods, demonstration of an antibody response to melanin in mice infected with C. neoformans and analysis of yeast cell walls in infected tissue by light microscopy, were used to support these findings. The demonstration that C. neoformans melanizes in tissue has important implications for pathogenesis and drug discovery. PMID- 9858596 TI - The E6 oncoproteins of high-risk papillomaviruses bind to a novel putative GAP protein, E6TP1, and target it for degradation. AB - The high-risk human papillomaviruses (HPVs) are associated with carcinomas of the cervix and other genital tumors. Previous studies have identified two viral oncoproteins, E6 and E7, which are expressed in the majority of HPV-associated carcinomas. The ability of high-risk HPV E6 protein to immortalize human mammary epithelial cells (MECs) has provided a single-gene model to study the mechanisms of E6-induced oncogenic transformation. In this system, the E6 protein targets the p53 tumor suppressor protein for degradation, and mutational analyses have shown that E6-induced degradation of p53 protein is required for MEC immortalization. However, the inability of most dominant-negative p53 mutants to induce efficient immortalization of MECs suggests the existence of additional targets of the HPV E6 oncoprotein. Using the yeast two-hybrid system, we have isolated a novel E6-binding protein. This polypeptide, designated E6TP1 (E6 targeted protein 1), exhibits high homology to GTPase-activating proteins for Rap, including SPA-1, tuberin, and Rap1GAP. The mRNA for E6TP1 is widely expressed in tissues and in vitro-cultured cell lines. The gene for E6TP1 localizes to chromosome 14q23.2-14q24.3 within a locus that has been shown to undergo loss of heterozygosity in malignant meningiomas. Importantly, E6TP1 is targeted for degradation by the high-risk but not the low-risk HPV E6 proteins both in vitro and in vivo. Furthermore, the immortalization-competent but not the immortalization-incompetent HPV16 E6 mutants target the E6TP1 protein for degradation. Our results identify a novel target for the E6 oncoprotein and provide a potential link between HPV E6 oncogenesis and alteration of a small G protein signaling pathway. PMID- 9858598 TI - Withdrawal of survival factors results in activation of the JNK pathway in neuronal cells leading to Fas ligand induction and cell death. AB - The JNK pathway modulates AP-1 activity. While in some cells it may have proliferative and protective roles, in neuronal cells it is involved in apoptosis in response to stress or withdrawal of survival signals. To understand how JNK activation leads to apoptosis, we used PC12 cells and primary neuronal cultures. In PC12 cells, deliberate JNK activation is followed by induction of Fas ligand (FasL) expression and apoptosis. JNK activation detected by c-Jun phosphorylation and FasL induction are also observed after removal of either nerve growth factor from differentiated PC12 cells or KCl from primary cerebellar granule neurons (CGCs). Sequestation of FasL by incubation with a Fas-Fc decoy inhibits apoptosis in all three cases. CGCs derived from gld mice (defective in FasL) are less sensitive to apoptosis caused by KCl removal than wild-type neurons. In PC12 cells, protection is also conferred by a c-Jun mutant lacking JNK phosphoacceptor sites and a small molecule inhibitor of p38 mitogen-activated protein kinase and JNK, which inhibits FasL induction. Hence, the JNK-to-c-Jun-to-FasL pathway is an important mediator of stress-induced neuronal apoptosis. PMID- 9858599 TI - CREB binding protein interacts with nucleoporin-specific FG repeats that activate transcription and mediate NUP98-HOXA9 oncogenicity. AB - Genes encoding the Phe-Gly (FG) repeat-containing nucleoporins NUP98 and CAN/NUP214 are at the breakpoints of several chromosomal translocations associated with human acute myeloid leukemia (AML), but their role in oncogenesis is unclear. Here we demonstrate that the NUP98-HOXA9 fusion gene encodes two nuclear oncoproteins with either 19 or 37 NUP98 FG repeats fused to the DNA binding and PBX heterodimerization domains of the transcription factor HOXA9. Both NUP98-HOXA9 chimeras transformed NIH 3T3 fibroblasts, and this transformation required the HOXA9 domains for DNA binding and PBX interaction. Surprisingly, the FG repeats acted as very potent transactivators of gene transcription. This NUP98-derived activity is essential for transformation and can be replaced by the bona fide transactivation domain of VP16. Interestingly, FG repeat-containing segments derived from the nucleoporins NUP153 and CAN/NUP214 functioned similarly to those from NUP98. We further demonstrate that transactivation by FG repeat-rich segments of NUP98 correlates with their ability to interact functionally and physically with the transcriptional coactivators CREB binding protein (CBP) and p300. This finding shows, for the first time, that a translocation-generated fusion protein appears to recruit CBP/p300 as an important step of its oncogenic mechanism. Together, our results suggest that NUP98-HOXA9 chimeras are aberrant transcription factors that deregulate HOX responsive genes through the transcriptional activation properties of nucleoporin specific FG repeats that recruit CBP/p300. Indeed, FG repeat-mediated transactivation may be a shared pathogenic function of nucleoporins implicated human AML. PMID- 9858600 TI - C-Terminal binding protein is a transcriptional repressor that interacts with a specific class of vertebrate Polycomb proteins. AB - Polycomb (Pc) is part of a Pc group (PcG) protein complex that is involved in repression of gene activity during Drosophila and vertebrate development. To identify proteins that interact with vertebrate Pc homologs, we performed two hybrid screens with Xenopus Pc (XPc) and human Pc 2 (HPC2). We find that the C terminal binding protein (CtBP) interacts with XPc and HPC2, that CtBP and HPC2 coimmunoprecipitate, and that CtBP and HPC2 partially colocalize in large PcG domains in interphase nuclei. CtBP is a protein with unknown function that binds to a conserved 6-amino-acid motif in the C terminus of the adenovirus E1A protein. Also, the Drosophila CtBP homolog interacts, through this conserved amino acid motif, with several segmentation proteins that act as repressors. Similarly, we find that CtBP binds with HPC2 and XPc through the conserved 6 amino-acid motif. Importantly, CtBP does not interact with another vertebrate Pc homolog, M33, which lacks this amino acid motif, indicating specificity among vertebrate Pc homologs. Finally, we show that CtBP is a transcriptional repressor. The results are discussed in terms of a model that brings together PcG mediated repression and repression systems that require corepressors such as CtBP. PMID- 9858601 TI - Human osteogenesis involves differentiation-dependent increases in the morphogenically active 3' alternative splicing variant of acetylcholinesterase. AB - The extended human acetylcholinesterase (AChE) promoter contains many binding sites for osteogenic factors, including 1,25-(OH)2 vitamin D3 and 17beta estradiol. In differentiating osteosarcoma Saos-2 cells, both of these factors enhanced transcription of the AChE mRNA variant 3' terminated with exon 6 (E6 AChE mRNA), which encodes the catalytically and morphogenically active E6-AChE isoform. In contrast, antisense oligodeoxynucleotide suppression of E6-AChE mRNA expression increased Saos-2 proliferation in a dose- and sequence-dependent manner. The antisense mechanism of action was most likely mediated by mRNA destruction or translational arrest, as cytochemical staining revealed reduction in AChE gene expression. In vivo, we found that E6-AChE mRNA levels rose following midgestation in normally differentiating, postproliferative fetal chondrocytes but not in the osteogenically impaired chondrocytes of dwarf fetuses with thanatophoric dysplasia. Taken together, these findings suggest morphogenic involvement of E6-AChE in the proliferation-differentiation balance characteristic of human osteogenesis. PMID- 9858602 TI - Histone acetyltransferase and protein kinase activities copurify with a putative Xenopus RNA polymerase I holoenzyme self-sufficient for promoter-dependent transcription. AB - Mounting evidence suggests that eukaryotic RNA polymerases preassociate with multiple transcription factors in the absence of DNA, forming RNA polymerase holoenzyme complexes. We have purified an apparent RNA polymerase I (Pol I) holoenzyme from Xenopus laevis cells by sequential chromatography on five columns: DEAE-Sepharose, Biorex 70, Sephacryl S300, Mono Q, and DNA-cellulose. Single fractions from every column programmed accurate promoter-dependent transcription. Upon gel filtration chromatography, the Pol I holoenzyme elutes at a position overlapping the peak of Blue Dextran, suggesting a molecular mass in the range of approximately 2 MDa. Consistent with its large mass, Coomassie blue stained sodium dodecyl sulfate-polyacrylamide gels reveal approximately 55 proteins in fractions purified to near homogeneity. Western blotting shows that TATA-binding protein precisely copurifies with holoenzyme activity, whereas the abundant Pol I transactivator upstream binding factor does not. Also copurifying with the holoenzyme are casein kinase II and a histone acetyltransferase activity with a substrate preference for histone H3. These results extend to Pol I the suggestion that signal transduction and chromatin-modifying activities are associated with eukaryotic RNA polymerases. PMID- 9858603 TI - Ribosomal pausing and scanning arrest as mechanisms of translational regulation from cap-distal iron-responsive elements. AB - Iron regulatory protein 1 (IRP-1) binding to an iron-responsive element (IRE) located close to the cap structure of mRNAs represses translation by precluding the recruitment of the small ribosomal subunit to these mRNAs. This mechanism is position dependent; reporter mRNAs bearing IREs located further downstream exhibit diminished translational control in transfected mammalian cells. To investigate the underlying mechanism, we have recapitulated this position effect in a rabbit reticulocyte cell-free translation system. We show that the recruitment of the 43S preinitiation complex to the mRNA is unaffected when IRP-1 is bound to a cap-distal IRE. Following 43S complex recruitment, the translation initiation apparatus appears to stall, before linearly progressing to the initiation codon. The slow passive dissociation rate of IRP-1 from the cap-distal IRE suggests that the mammalian translation apparatus plays an active role in overcoming the cap-distal IRE-IRP-1 complex. In contrast, cap-distal IRE-IRP-1 complexes efficiently repress translation in wheat germ and yeast translation extracts. Since inhibition occurs subsequent to 43S complex recruitment, an efficient arrest of productive scanning may represent a second mechanism by which RNA-protein interactions within the 5' untranslated region of an mRNA can regulate translation. In contrast to initiating ribosomes, elongating ribosomes from mammal, plant, and yeast cells are unaffected by IRE-IRP-1 complexes positioned within the open reading frame. These data shed light on a characteristic aspect of the IRE-IRP regulatory system and uncover properties of the initiation and elongation translation apparatus of eukaryotic cells. PMID- 9858604 TI - The two forms of karyogamy transcription factor Kar4p are regulated by differential initiation of transcription, translation, and protein turnover. AB - Kar4p is a transcription factor in Saccharomyces cerevisiae that is required for the expression of karyogamy-specific genes during mating, for the efficient transit from G1 during mitosis, and for essential functions during meiosis. Kar4p exists in two forms: a constitutive slower-migrating form, which predominates during vegetative growth, and a faster-migrating form, which is highly induced by mating pheromone. Transcript mapping of KAR4 revealed that the constitutive mRNA was initiated upstream of two in-frame ATG initiation codons, while the major inducible mRNA originated between them. Thus, the two forms of Kar4p are derived from the translation of alternative transcripts, which possess different AUG initiation codons. Site-directed mutations were constructed to inactivate one or the other of the initiation codons, allowing the expression of the two Kar4p forms separately. At normal levels of expression, the constitutive form of Kar4p did not support wild-type levels of mating. However, the two forms of Kar4p could also be expressed separately from the regulatable GAL1 promoter, and no functional difference was detected when they were expressed at equivalent levels. Pulse-chase experiments showed that the induced form of Kar4p was highly expressed and stable during mating but rapidly turned over in vegetative cells. In contrast, the constitutively expressed longer form showed the same rate of turnover regardless of the growth condition. Furthermore, overexpression of either form of Kar4p in vegetative cells was toxic. Thus, the elaborate regulation of the two forms of Kar4p at the levels of transcription, translation, and protein turnover reflects the requirement for high levels of the protein during mating and for low levels during the subsequent phases of the cell cycle. PMID- 9858605 TI - Ribosomal protein S14 of Saccharomyces cerevisiae regulates its expression by binding to RPS14B pre-mRNA and to 18S rRNA. AB - Production of ribosomal protein S14 in Saccharomyces cerevisiae is coordinated with the rate of ribosome assembly by a feedback mechanism that represses expression of RPS14B. Three-hybrid assays in vivo and filter binding assays in vitro demonstrate that rpS14 directly binds to an RNA stem-loop structure in RPS14B pre-mRNA that is necessary for RPS14B regulation. Moreover, rpS14 binds to a conserved helix in 18S rRNA with approximately five- to sixfold-greater affinity. These results support the model that RPS14B regulation is mediated by direct binding of rpS14 either to its pre-mRNA or to rRNA. Investigation of these interactions with the three-hybrid system reveals two regions of rpS14 that are involved in RNA recognition. D52G and E55G mutations in rpS14 alter the specificity of rpS14 for RNA, as indicated by increased affinity for RPS14B RNA but reduced affinity for the rRNA target. Deletion of the C terminus of rpS14, where multiple antibiotic resistance mutations map, prevents binding of rpS14 to RNA and production of functional 40S subunits. The emetine-resistant protein, rpS14-EmRR, which contains two mutations near the C terminus of rpS14, does not bind either RNA target in the three-hybrid or in vitro assays. This is the first direct demonstration that an antibiotic resistance mutation alters binding of an r protein to rRNA and is consistent with the hypothesis that antibiotic resistance mutations can result from local alterations in rRNA structure. PMID- 9858606 TI - Two Xenopus proteins that bind the 3' end of histone mRNA: implications for translational control of histone synthesis during oogenesis. AB - Translationally inactive histone mRNA is stored in frog oocytes, and translation is activated at oocyte maturation. The replication-dependent histone mRNAs are not polyadenylated and end in a conserved stem-loop structure. There are two proteins (SLBPs) which bind the 3' end of histone mRNA in frog oocytes. SLBP1 participates in pre-mRNA processing in the nucleus. SLBP2 is oocyte specific, is present in the cytoplasm, and does not support pre-mRNA processing in vivo or in vitro. The stored histone mRNA is bound to SLBP2. As oocytes mature, SLBP2 is degraded and a larger fraction of the histone mRNA is bound to SLBP1. The mechanism of activation of translation of histone mRNAs may involve exchange of SLBPs associated with the 3' end of histone mRNA. PMID- 9858607 TI - Rb inhibits the intrinsic kinase activity of TATA-binding protein-associated factor TAFII250. AB - The retinoblastoma tumor suppressor protein, Rb, interacts directly with the largest TATA-binding protein-associated factor, TAFII250, through multiple regions in each protein. To define the potential role(s) of this interaction, we examined whether Rb could regulate the intrinsic, bipartite kinase activity of TAFII250. Here, we report that Rb is able to inhibit the kinase activity of immunopurified and gel-purified recombinant TAFII250. Rb inhibits the autophosphorylation of TAFII250 as well as its phosphorylation of the RAP74 subunit of TFIIF in a dose-responsive manner. Inhibition of TAFII250 kinase activity involves the Rb pocket (amino acids 379 to 928) but not its amino terminus. In addition, Rb appears to specifically inhibit the amino-terminal kinase domain of TAFII250 through a direct protein-protein interaction. We further demonstrate that two different tumor-derived Rb pocket mutants, C706F and Deltaex22, are functionally defective for kinase inhibition, even though they are able to bind the amino terminus of TAFII250. Our results suggest a novel mechanism of transcriptional regulation by Rb, involving direct interaction with TAFII250 and inhibition of its ability to phosphorylate itself, RAP74, and possibly other targets. PMID- 9858608 TI - Activation domain-specific and general transcription stimulation by native histone acetyltransferase complexes. AB - Recent progress in identifying the catalytic subunits of histone acetyltransferase (HAT) complexes has implicated histone acetylation in the regulation of transcription. Here, we have analyzed the function of two native yeast HAT complexes, SAGA (Spt-Ada-Gcn5 Acetyltransferase) and NuA4 (nucleosome acetyltransferase of H4), in activating transcription from preassembled nucleosomal array templates in vitro. Each complex was tested for the ability to enhance transcription driven by GAL4 derivatives containing either acidic, glutamine-rich, or proline-rich activation domains. On nucleosomal array templates, the SAGA complex selectively stimulates transcription driven by the VP16 acidic activation domain in an acetyl coenzyme A-dependent manner. In contrast, the NuA4 complex facilitates transcription mediated by any of the activation domains tested if allowed to preacetylate the nucleosomal template, indicating a general stimulatory effect of histone H4 acetylation. However, when the extent of acetylation by NuA4 is limited, the complex also preferentially stimulates VP16-driven transcription. SAGA and NuA4 interact directly with the VP16 activation domain but not with a glutamine-rich or proline-rich activation domain. These data suggest that recruitment of the SAGA and NuA4 HAT complexes by the VP16 activation domain contributes to HAT-dependent activation. In addition, extensive H4/H2B acetylation by NuA4 leads to a general activation of transcription, which is independent of activator-NuA4 interactions. PMID- 9858610 TI - The two Drosophila telomeric transposable elements have very different patterns of transcription. AB - The transposable elements HeT-A and TART constitute the telomeres of Drosophila chromosomes. Both are non-long terminal repeat (LTR) retrotransposons, sharing the remarkable property of transposing only to chromosome ends. In addition, strong sequence similarity of their gag proteins indicates that these coding regions share a common ancestor. These findings led to the assumption that HeT-A and TART are closely related. However, we now find that these elements produce quite different sets of transcripts. HeT-A produces only sense-strand transcripts of the full-length element, whereas TART produces both sense and antisense full length RNAs, with antisense transcripts in more than 10-fold excess over sense RNA. In addition, features of TART sequence organization resemble those of a subclass of non-LTR elements characterized by unequal terminal repeats. Thus, the ancestral gag sequence appears to have become incorporated in two different types of elements, possibly with different functions in the telomere. HeT-A transcripts are found in both nuclear and cytoplasmic cell fractions, consistent with roles as both mRNA and transposition template. In contrast, both sense and antisense TART transcripts are almost entirely concentrated in nuclear fractions. Also, TART open reading frame 2 probes detect a cytoplasmic mRNA for reverse transcriptase (RT), with no similarity to TART sequence 5' or 3' of the RT coding region. This RNA could be a processed TART transcript or the product of a "free standing" RT gene. Either origin would be novel. The distinctive transcription patterns of both HeT-A and TART are conserved in Drosophila yakuba, despite significant sequence divergence. The conservation argues that these sets of transcripts are important to the function(s) of HeT-A and TART. PMID- 9858609 TI - Regulation of the mdm2 oncogene by thyroid hormone receptor. AB - The mdm2 gene is positively regulated by p53 through a p53-responsive DNA element in the first intron of the mdm2 gene. mdm2 binds p53, thereby abrogating the ability of p53 to activate the mdm2 gene, and thus forming an autoregulatory loop of mdm2 gene regulation. Although the mdm2 gene is thought to act as an oncogene by blocking the activity of p53, recent studies indicate that mdm2 can act independently of p53 and block the G1 cell cycle arrest mediated by members of the retinoblastoma gene family and can activate E2F1/DP1 and the cyclin A gene promoter. In addition, factors other than p53 have recently been shown to regulate the mdm2 gene. In this article, we report that thyroid hormone (T3) receptors (T3Rs), but not the closely related members of the nuclear thyroid hormone/retinoid receptor gene family (retinoic acid receptor, vitamin D receptor, peroxisome proliferation activation receptor, or retinoid X receptor), regulate mdm2 through the same intron sequences that are modulated by p53. Chicken ovalbumin upstream promoter transcription factor I, an orphan nuclear receptor which normally acts as a transcriptional repressor, also activates mdm2 through the same intron region of the mdm2 gene. Two T3R-responsive DNA elements were identified and further mapped to sequences within each of the p53 binding sites of the mdm2 intron. A 10-amino-acid sequence in the N-terminal region of T3Ralpha that is important for transactivation and interaction with TFIIB was also found to be important for activation of the mdm2 gene response element. T3 was found to stimulate the endogenous mdm2 gene in GH4C1 cells. These cells are known to express T3Rs, and T3 is known to stimulate replication of these cells via an effect in the G1 phase of the cell cycle. Our findings, which indicate that T3Rs can regulate the mdm2 gene independently of p53, provide an explanation for certain known effects of T3 and T3Rs on cell proliferation. In addition, these findings provide further evidence for p53-independent regulation of mdm2 which could lead to the development of tumors from cells that express low levels of p53 or that express p53 mutants defective in binding to and activating the mdm2 gene. PMID- 9858611 TI - Activation of transcription by metabolic intermediates of the pyrimidine biosynthetic pathway. AB - Saccharomyces cerevisiae responds to pyrimidine starvation by increasing the expression of four URA genes, encoding the enzymes of de novo pyrimidine biosynthesis, three- to eightfold. The increase in gene expression is dependent on a transcriptional activator protein, Ppr1p. Here, we investigate the mechanism by which the transcriptional activity of Ppr1p responds to the level of pyrimidine biosynthetic intermediates. We find that purified Ppr1p is unable to promote activation of transcription in an in vitro system. Transcriptional activation by Ppr1p can be observed, however, if either dihydroorotic acid (DHO) or orotic acid (OA) is included in the transcription reactions. The transcriptional activation function and the DHO/OA-responsive element of Ppr1p localize to the carboxyl-terminal 134 amino acids of the protein. Thus, Ppr1p directly senses the level of early pyrimidine biosynthetic intermediates within the cell and activates the expression of genes encoding proteins required later in the pathway. These results are discussed in terms of (i) regulation of the pyrimidine biosynthetic pathway and (ii) a novel mechanism of regulating gene expression. PMID- 9858612 TI - Cyclosporin A promotes translational silencing of autocrine interleukin-3 via ribosome-associated deadenylation. AB - Translation is regulated predominantly by an interplay between cis elements at the 3' and 5' ends of mRNAs and trans-acting proteins. Cyclosporin A (CsA), a calcineurin antagonist and blocker of interleukin-2 (IL-2) transcription in T cells, was found to inhibit translation of IL-3 mRNA in autocrine mast cell tumor lines. The mechanism involved ribosome-associated poly(A) shortening and required an intact AU-rich element in the 3' untranslated region. FK506, another calcineurin inhibitor, shared the effect. The translational inhibition by CsA was specific to oncogenically induced lymphokines IL-3 and IL-4 but not to IL-6, c jun, and c-myc, which are expressed in the nonmalignant precursor cells. Furthermore, no translational down-regulation of the mRNA was observed in IL-3 transfected precursor cells. These data suggest that translational silencing is associated with the tumor phenotype. PMID- 9858613 TI - Coactivator PC4 mediates AP-2 transcriptional activity and suppresses ras-induced transformation dependent on AP-2 transcriptional interference. AB - ras oncogene-transformed PA-1 human teratocarcinoma cells have abundant AP-2 mRNA but, paradoxically, little AP-2 transcriptional activity. We have previously shown that overexpression of AP-2 in nontumorigenic variants of PA-1 cells results in inhibition of AP-2 activity and induction of tumorigenicity similar to that caused by ras transformation of PA-1 cells. Evidence indicated the existence of a novel mechanism of inhibition of AP-2 activity involving sequestering of transcriptional coactivators. In this study, we found that PC4 is a positive coactivator of AP-2 and can restore AP-2 activity in ras-transformed PA-1 cells. Relative to vector-transfected ras cell lines, ras cell lines stably transfected with and expressing the PC4 cDNA have a diminished growth rate and exhibit a loss of anchorage-independent growth, and they are unable to induce the formation of tumors in nude mice. These data suggest that a transcriptional coactivator, like a tumor suppressor, can have a growth-suppressive effect on cells. Our experiments are the first to show that ras oncogenes and oncogenic transcription factors can induce transformation through effects on the transcription machinery rather than through specific programs of gene expression. PMID- 9858614 TI - Selected elements of herpes simplex virus accessory factor HCF are highly conserved in Caenorhabditis elegans. AB - HCF is a mammalian nuclear protein that undergoes proteolytic processing and is required for cell proliferation. During productive herpes simplex virus (HSV) infection, the viral transactivator VP16 associates with HCF to initiate HSV gene transcription. Here, we show that the worm Caenorhabditis elegans possesses a functional homolog of mammalian HCF that can associate with and activate the viral protein VP16. The pattern of sequence conservation, however, is uneven. Sequences required for mammalian HCF processing are not present in C. elegans HCF. Furthermore, not all elements of mammalian HCF that are required for promoting cell proliferation are conserved. Nevertheless, unexpectedly, C. elegans HCF can promote mammalian cell proliferation because a region of HCF that is conserved can promote mammalian cell proliferation better than its human counterpart. These results suggest that HCF possesses a highly conserved role in metazoan cell proliferation which is targeted by VP16 to regulate HSV infection. The precise mechanisms, however, by which HCF functions in mammals and worms appear to differ. PMID- 9858615 TI - E2F and histone deacetylase mediate transforming growth factor beta repression of cdc25A during keratinocyte cell cycle arrest. AB - cdc25A is a tyrosine phosphatase that activates G1 cyclin-dependent kinases (Cdk's). In human keratinocytes, cdc25A expression is down-regulated after the initial drop in Cdk activity caused by cell exposure to the antimitogenic cytokine transforming growth factor beta (TGF-beta) or removal of serum factors. Here we show that the TGF-beta-inhibitory-response element in the cdc25A promoter maps to an E2F site at nucleotides -62 to -55 from the transcription start site. This site is not required for basal transcription in keratinocytes. We provide evidence that the cell cycle arrest program activated by TGF-beta in human keratinocytes includes the generation of E2F4-p130 complexes that in association with histone deacetylase HDAC1 inhibit the activity of the cdc25A promoter from this repressor E2F site. This mechanism is part of a program that places keratinocytes in the quiescent state following the initial drop in Cdk activity caused by cell exposure to TGF-beta. PMID- 9858616 TI - Gbp1p, a protein with RNA recognition motifs, binds single-stranded telomeric DNA and changes its binding specificity upon dimerization. AB - Gbp1p is a putative telomere-binding protein from Chlamydomonas reinhardtii that contains two RNA recognition motifs (RRMs) which are commonly found in heterogeneous nuclear ribonucleoproteins (hnRNPs). Previously we demonstrated that Gbp1p binds single-stranded DNA (ssDNA) containing the Chlamydomonas telomeric sequence but not the RNA containing the cognate sequence. Here we show that at lower protein concentrations Gbp1 can also bind an RNA containing the cognate sequence. We found that mutation of the two RRM motifs of Gbp1p to match the highly conserved region of hnRNP RRMs did not alter the affinity of Gbp1p for either RNA or DNA. The ability of Gbp1p to associate with either of these two nucleic acids is governed by the dimerization state of the protein. Monomeric Gbp1p associates with either ssDNA or RNA, showing a small binding preference for RNA. Dimeric Gbp1p has a strong preference for binding ssDNA and shows little affinity for RNA. To the best of our knowledge, this is the first example of a protein that qualitatively shifts its nucleic acid binding preference upon dimerization. The biological implications of a telomere-binding protein that is regulated by dimerization are discussed. PMID- 9858617 TI - RNA polymerase II transcription suppresses nucleosomal modulation of UV-induced (6-4) photoproduct and cyclobutane pyrimidine dimer repair in yeast. AB - The nucleotide excision repair (NER) pathway is able to remove a wide variety of structurally unrelated lesions from DNA. NER operates throughout the genome, but the efficiencies of lesion removal are not the same for different genomic regions. Even within a single gene or DNA strand repair rates vary, and this intragenic heterogeneity is of considerable interest with respect to the mutagenic potential of carcinogens. In this study, we have analyzed the removal of the two major types of genotoxic DNA adducts induced by UV light, i.e., the pyrimidine (6-4)-pyrimidone photoproduct (6-4PP) and the cyclobutane pyrimidine dimer (CPD), from the Saccharomyces cerevisiae URA3 gene at nucleotide resolution. In contrast to the fast and uniform removal of CPDs from the transcribed strand, removal of lesions from the nontranscribed strand is generally less efficient and is modulated by the chromatin environment of the damage. Removal of 6-4PPs from nontranscribed sequences is also profoundly influenced by positioned nucleosomes, but this type of lesion is repaired at a much higher rate. Still, the transcribed strand is repaired preferentially, indicating that, as in the removal of CPDs, transcription-coupled repair predominates in the removal of 6-4PPs from transcribed DNA. The hypothesis that transcription machinery operates as the rate-determining damage recognition entity in transcription-coupled repair is supported by the observation that this pathway removes both types of UV photoproducts at equal rates without being profoundly influenced by the sequence or chromatin context. PMID- 9858620 TI - Editorial AB - In this issue of the Journal of Helminthology I have much pleasure in including a selection of papers drawn from two workshops, one on 'Nematode Population Genetics' and another on the 'Ecology of Fish Parasites' held in April 1998 at Exeter during the Spring Meeting of the British Society for Parasitology (BSP).The aim of the workshop on 'Nematode Population Genetics' convened by Dr Mark Viney, University of Edinburgh, is to review the current state of knowledge of this topic and to consider future research priorities. Following Mark Viney's overview, Michael Blouin, Oregon State University, Corvallis, USA, considers the effects of parasite life histories on mitochondrial DNA diversity in nematodes. Marleen Roos et al., Institute for Animal Science and Health and the University of Utrecht, The Netherlands, then review polymorphic DNA markers in the genome of parasitic nematodes whereas in the fourth and final paper of this series Alison Galvani and Sunetra Gupta, University of Oxford, consider the effects of mating probability on the population genetics of nematodes.Two papers from the workshop on the 'Ecology of Fish Parasites', convened by Professor Clive Kennedy, University of Exeter, focus on the community ecology of helminths in fish and include a study by Clive Kennedy et al., University of Exeter and University of Rome on the 'Composition and diversity of helminth communities in eels in the River Tiber: long term changes and comparison with insular Europe'. This is followed by an invited paper by William Font, Southeastern Louisiana University, USA on 'Parasites in paradise: patterns of helminth distribution in Hawaiian stream fishes'.I wish to express my grateful thanks to Mark Viney and Clive Kennedy for their assistance in the publication of these papers for this special issue. PMID- 9858618 TI - Interactions between the class II transactivator and CREB binding protein increase transcription of major histocompatibility complex class II genes. AB - Class II major histocompatibility (class II) genes are regulated in a B-cell specific and gamma interferon-inducible fashion. The master switch for the expression of these genes is the class II transactivator (CIITA). In this report, we demonstrate that one of the functions of CIITA is to recruit the CREB binding protein (CBP) to class II promoters. Not only functional but also specific binding interactions between CIITA and CBP were demonstrated. Moreover, a dominant negative form of CBP decreased the activity of class II promoters and levels of class II determinants on the surface of cells. Finally, the inhibition of class II gene expression by the glucocorticoid hormone could be attributed to the squelching of CBP by the glucocorticoid receptor. We conclude that CBP, a histone acetyltransferase, plays an important role in the transcription of class II genes. PMID- 9858621 TI - Nematode population genetics. PMID- 9858622 TI - Mitochondrial DNA diversity in nematodes. AB - The relatively small literature on mitochondrial DNA (mtDNA) diversity in nematode species is summarized here. Nematodes show a wide range of overall genetic diversities and population genetic structures. Species-wide levels of diversity correlate strongly with the breeding system and other life cycle features that control effective population size. Obligate outcrossers that parasitize mobile vertebrate hosts are the most diverse, species having hermaphroditic stages are less so, and species having asexual reproductive stages appear even less diverse. Nevertheless, these conclusions are preliminary because there exist so few data on DNA diversity in nematodes. What is needed are more comparative studies using similar sampling designs and the same DNA markers, including nuclear loci and further work with mtDNA. PMID- 9858619 TI - Interdomain B in ZAP-70 regulates but is not required for ZAP-70 signaling function in lymphocytes. AB - The protein tyrosine kinase ZAP-70 plays an important role in T-cell activation and development. After T-cell receptor stimulation, ZAP-70 associates with the receptor and is phosphorylated on many tyrosines, including Y292, Y315, and Y319 within interdomain B. Previously, we demonstrated that Y292 negatively regulates ZAP-70 function and that Y315 positively regulates ZAP-70 function by interacting with Vav. Recent studies have suggested that Y319 also positively regulate ZAP-70 function. Paradoxically, removal of interdomain B (to create the construct designated Delta), containing the Y292, Y315, and Y319 sites, did not eliminate the ability of ZAP-70 to induce multiple gene reporters in Syk-deficient DT-40 B cells and ZAP-70/Syk-deficient Jurkat cells. Here we show that Delta still utilizes the same pathways as wild-type ZAP-70 to mediate NF-AT induction. This is manifested by the ability of Delta to restore induction of calcium fluxes and mitogen-activated protein kinase activation and by the ability of dominant negative Ras and FK506 to block the induction of NF-AT activity mediated by Delta. Biochemically we show that the stimulated tyrosine phosphorylation of Vav, Shc, and ZAP-70 itself is diminished, whereas that of Slp-76 is increased in cells reconstituted with Delta. Deletion of interdomain B did not affect the ability of ZAP-70 to bind to the receptor. The in vitro kinase activity of ZAP-70 lacking interdomain B was markedly reduced, but the kinase activity was still required for the protein's in vivo activity. Based on these data, we concluded that interdomain B regulates but is not required for ZAP-70 signaling function leading to cellular responses. PMID- 9858623 TI - Polymorphic DNA markers in the genome of parasitic nematodes. AB - Polymorphic molecular markers are being identified to characterize the genomes of parasitic nematodes. The aim is to construct a map with markers evenly spread over the six chromosomes. With such a map, regions can be identified that are under selection pressure when attempts are being made to eradicate worms, be it by drugs, vaccines or genetic resistance in the sheep. Several types of markers have been developed, microsatellites, transposon-associated markers, amplified fragment length polymorphism (AFLP) and expressed sequence tag (EST) markers. Linkage groups can be constructed using several genetic crosses between inbred and drug resistant strains. EST markers will be especially important for comparative mapping with the genome of Caenorhabditis elegans, and therefore localization of the linkage group on a chromosome. It will then be possible to identify functional genes close to markers that have changed allele frequencies under selection pressure and identify the mechanisms of resistance to parasite control. PMID- 9858624 TI - The effects of mating probability on the population genetics of nematodes. AB - We review how constraints on the mating probability of female worms by segregation within individual host guts form a critical element in the parasite population structure of nematodes. We consider the effects of these constraints on the population genetics of nematodes under various assumptions regarding worm competition as reflected in the relationship between the abundance of a certain genotype within the gut and the probability of reproductive success. The consequences for the emergence of resistance to drugs and immunotherapy and implications for host-parasite coevolution are discussed. We also review evidence for genetic heterogeneity in parasite populations as a necessary prerequisite for the applicability of mating probability models designed to assess the population genetics of nematodes. PMID- 9858625 TI - Composition and diversity of helminth communities in eels anguilla anguilla in the river tiber: long-term changes and comparison with insular europe AB - Most studies of helminth communities in the European eel Anguilla anguilla have been undertaken in the British Isles, and there are very few analyses of community composition and structure from continental Europe. To fill this gap and test the hypothesis that helminth communities in freshwater eels in the British Isles are not typical of those of continental Europe, helminth communities of eels in the River Tiber below Rome were analysed by season using data collected in 1980 and new data from 1996. The intestinal helminth communities in the Tiber eels were species poor and characterized by low diversity. Most eels harboured one or no parasite species and communities were heavily dominated by the acanthocephalan Acanthocephalus clavula. Intestinal helminth infracommunity richness and diversity did not differ between seasons within a year or between the same seasons in 1980 and 1996, although some changes in composition were apparent. Intestinal infracommunities from Tiber eels were very similar in characteristics to those analysed from the British Isles, and their temporal changes also showed close similarities to those reported from rivers in the UK. It seems likely therefore that conclusions derived from British studies can be applied to helminth communities of eels on the continent. PMID- 9858626 TI - Parasites in paradise: patterns of helminth distribution in hawaiian stream fishes AB - Of the 13 species of helminths that parasitize stream fishes in Hawai'i, seven species are considered to be native to the archipelago and the remaining six species to be introduced by man. Sources of colonization for native species are piscivorous birds for three species, and marine fishes for four species. Non native helminths have been brought to Hawai'i in association with the importation of parasitized exotic species of poeciliids introduced into streams for mosquito control and as aquarium releases. Many of these introduced parasites have broad host specificity and now infect the five species of native gobioid stream fishes. Exotic parasites, including a roundworm Camallanus cotti, a tapeworm Bothriocephalus acheilognathi and a leech Myzobdella lugubris, are more widely distributed among Hawaiian streams than are native species. PMID- 9858627 TI - Morphological variability in Fasciola hepatica eggs in ruminants, rodents and lagomorphs. AB - The length and width of 1297 Fasciola hepatica eggs shed in cattle hosts, 337 in sheep and 199 in nutria, were measured from several parts of France. The data were compared with those obtained from other studies in Spain, France (where rats were also investigated), Germany and the Netherlands. One way analysis of variance and discriminant analysis were used to assess differences between host origins. The distribution of length and width of eggs were analysed using skewness and kurtosis Fisher coefficients. The eggs recovered from sheep, cattle, rodents and lagomorphs were different in size: the eggs found in rodents (length L x width W in microm: 8592) and lagomorphs (L x W in microm: 9100) were smaller than those found in sheep and cattle (L x W in microm: 10,000). These morphological differences in F. hepatica eggs were host-induced in rats (L x W in microM: 9709 in cattle to 8949 in rats) and rabbits (L x W in microm: 9709 in cattle to 8432 in rabbits). These differences in size of eggs might correspond to their being less able to develop into miracidia in less frequent hosts such as rodents and rabbits. PMID- 9858628 TI - Population dynamics of and larval trematode interactions with Lymnaea tomentosa and the potential for biological control of schistosome dermatitis in Bremner Bay, Lake Wanaka, New Zealand. AB - Lymnaea tomentosa, the intermediate host of a schistosome which causes schistosome dermatitis in Lake Wanaka, was found to depths of 16 m. The snail recruits in January, lives for up to 21 months, is associated with all water plant species and is found in areas devoid of macroscopic plants. It is host to several trematode species. Avian schistosome infections appear in the spring when echinostome prevalence is low, and a decrease in schistosome prevalence may be correlated with an increase in echinostome prevalence during the summer. A multiple-kind lottery model analysis of parasite species richness implies that interspecific interactions may be occurring in the host snail during the months of December and January. PMID- 9858629 TI - The effects of salinity, pH and temperature on the half-life and longevity of Echinostoma caproni miracidia. AB - Miracidia of Echinostoma caproni were exposed to solutions varying in salinity, pH, and temperature in 1 ml concavity slides. Half-lives of the miracidial populations were determined and longevity curves constructed to find maximum life spans of the miracidia in the different conditions. Control miracidia in aquarium water at pH 7.2 and 22 degrees C. had a half-life of 3.6 h and a maximum life span of 9 h. Miracidia of E. caproni were not very tolerant of saline solutions from 0.1% to 0.4%, the latter being lethal within an hour. A bimodal effect was found with exposure to aquarium water of varying pH, with a peak at pH 5 in acid solutions and pH 9 in alkaline solutions. Miracidia tolerated pH ranges from 3 to 11 exhibiting half-lives of 2.4 h or greater in these solutions. At lower than ambient temperatures, E. caproni miracidia lived longer, the greatest being a half-life of 5.0 h and a maximum life span of 15 h at 5 degrees C. At warmer temperatures, the half-life was reduced until lethality was reached at 40 degrees C. ANCOVA analysis of log transformed longevity curves supported the observation that in pH trials many miracidia survived initially with a major die-off after 3.3 h. The opposite trend of early die-off and gradual mortality of the survivors was supported in the temperature trials. PMID- 9858630 TI - Populations of acanthocephalus anguillaePomphorhynchus laevis in rivers with different pollution levels AB - The distribution of two acanthocephalan species (Pomphorhynchus laevisAcanthocephalus anguillae) in the chub (Leuciscus cephalus) was studied in four river reaches characterized by different levels of pollution: the River Ticino near Abbiategrasso (unpolluted), the Naviglio Grande Canal, in Milano (slightly polluted), the River Lambro near Merone village (polluted) and the River Lambro near Monza (severely polluted).Pomphorhynchus laevis was restricted to the unpolluted and the slightly polluted sites, while the intensity of A. anguillae increased proportionally to water pollution. These differences were partially explained by the variation in abundance of their intermediate hosts (Echinogammarus stammeri for P. laevisAsellus aquaticus for A. anguillae). Data on the occurrence of P. laevis and A. anguillae showed a significant negative binomial frequency distribution, suggesting their tendency to be aggregated within the host populations of L. cephalus. PMID- 9858631 TI - Morphological and developmental characteristics of Echinococcus granulosus derived from sheep, cattle and camels in Iran. AB - Echinococcus granulosus is widely distributed throughout Iran, where a variety of animals act as intermediate hosts. In this study, the development and morphological characteristics of E. granulosus of sheep, cattle and camel origin were compared and, overall, the characteristics of larval and adult isolates from sheep and cattle, although similar, showed significant differences from those of the camel isolates, especially with respect to the size of rostellar hooks. Differences in the fertility rates of hydatid cyst in sheep, cattle and camel were recorded as 88%, 19% and 70% respectively. In dogs experimentally infected with the three 'strains' of E. granulosus, the rate of development was more rapid in the camel than in the sheep and cattle 'strain'. In addition, adult E. granulosus derived from the camel demonstrate differences in worm length, segmentation and size, shape, number and arrangement of the genitalia when compared with worms derived from sheep and cattle. The epidemiological significance of these findings is discussed in relation to human hydatid infections in Iran. PMID- 9858632 TI - Biological control of Haemonchus contortus infective larvae in ovine faeces by administering an oral suspension of Duddingtonia flagrans chlamydospores to sheep. AB - A single oral dose of an aqueous suspension containing 11,350,000 chlamydospores of a Mexican isolate of Duddingtonia flagrans (FTHO-8) given to sheep, resulted in a maximum reduction of 88% (range 86.7-90.4%) of the population of Haemonchus contortus infective larvae in the faeces. The effect of this treatment continued for 4-5 days after administration of the suspension. The possible use of this treatment as a method of control of ovine haemonchosis is discussed. PMID- 9858633 TI - Morphological changes of larval Echinococcus multilocularis in mice treated with albendazole or mebendazole. AB - Using DBA/2J mice, tissue homogenates of larval Echinococcus multilocularis were injected into the mesenteric veins to generate the liver infection. Mice were treated with either albendazole or mebendazole for prolonged periods to examine the morphological changes of the metacestode. Albendazole induced disorganization of both laminated and germinal layers and suppressed the maturation of vesicles. Amorphous but loosely laminated PAS-positive material was observed inside the damaged vesicles, although new vesicles slightly developed inside or outside of the damaged ones. Active proliferation of vesicles occurred after treatment with albendazole was terminated. Hydatid cysts were more severely damaged in mice treated with mebendazole and new vesicles did not develop around the damaged ones. Also, hydatid cysts reappeared after treatment with mebendazole was terminated. These results indicate that these drugs do not eliminate larval E. multilocularis in the long-term, but mebendazole has a higher suppressive effect on multivesiculation than albendazole. PMID- 9858634 TI - Effects of Echinostoma caproni infection on the phospholipid and sphingolipid content of the intestinal mucosa of ICR mice. AB - High performance thin layer chromatography (HPTLC) was used to determine phospholipids and sphingolipids in the intestinal mucosa of ICR mice infected with Echinostoma caproni for two weeks. The major phospholipids detected in both infected and non-infected mucosa were phosphatidylcholine (PC) and phosphatidylethanolamine (PE). HPTLC-densitometric analysis showed that there was a significant decrease in the weight of both PC and PE in the intestinal mucosa of infected mice compared to that of the uninfected controls. Cerebrosides and sulphatides, but not sphingomyelin, were identified in the intestinal mucosa of both infected and uninfected hosts. There was an apparent increase in the cerebroside content of the mucosa of infected versus control mice. The pathobiochemical changes seen in the polar lipid content of infected hosts probably reflect the feeding and behavioural activities of E. caproni in the mouse intestine. PMID- 9858635 TI - Specific gravity of Opisthorchis viverrini eggs. AB - The specific gravity of the eggs of the liver fluke Opisthorchisviverrini was determined using a sucrose gradient centrifugation and found to range from 1.2713 to 1.3043. The peak egg count was located at the sucrose fraction with a specific gravity of 1.2814. An attempt to float eggs in saturated sodium nitrate solution, sp.gr. 1.4, failed. Examination of human stool specimens for Oviverrini eggs by simple flotation in saturated sodium nitrate solution and the formol-ether sedimentation technique revealed that the flotation technique was not as efficient as the sedimentation technique. It was suggested that the flotation techniques were inappropriate for the detection of Oviverrini eggs in faeces or contaminated soil. PMID- 9858636 TI - Heparan sulfate upregulates platelet-derived growth factor receptors on human lung fibroblasts. AB - Heparan sulfate is a molecule that possesses a large structural variability and which has been shown to inhibit the proliferation of fibroblasts in vitro. The aim of this study was to determine whether the anti-proliferative effects of heparan sulfate were exerted by regulation of the activity of the platelet derived growth factor and/or of the platelet-derived growth factor receptors. Both l-iduronate-rich, anti-proliferative and the l-iduronate-poor, non-anti proliferative heparan sulfate species, were incubated with confluent human embryonic lung fibroblasts for 24 h. The mRNA levels for PDGF-AA, PDGF-BB, and their receptors were measured. Binding studies were performed with [125I]-PDGF-BB and [125I]-EGF for 2 h at 4 degreesC in cultures preincubated with both types of heparan sulfate for 24 h. In separate experiments, cultures were incubated together with heparan sulfate and [125I]-PDGF-BB for 2 h at 4 degreesC. Increases of two- to threefold in the mRNA levels for both the alpha- and the beta receptors of PDGF was obtained after treatment with both types of heparan sulfate, whereas the mRNA levels of both the PDGF-AA and the PDGF-BB were essentially unaffected. A sixfold increase in binding was only noted for [125I] PDGF-BB in cultures pre-treated with the anti-proliferative heparan sulfate for 24 h, whereas no effect was noted with use of the non-anti-proliferative heparan sulfate. Incubating the [125I]-PDGF-BB and the anti-proliferative heparan sulfate together for 2 h resulted in a smaller, threefold increase in binding. This indicates that the anti-proliferative heparan sulfate both stabilizes and increases expression of the PDGF receptors. To investigate whether the increased number of PDGF receptors could affect cell activity, cells were preincubated with anti-proliferative heparan sulfate and then treated with PDGF-BB. This resulted in an increase in mitogenicity compared to cells treated only with PDGF-BB. Neither an increase in binding for [125I-EGF] nor an increase in the mitogenic response of EGF could be observed in cultures pre-treated with the anti proliferative heparan sulfate. The results indicate that the extracellular matrix itself may regulate important biological phenomena such as cell proliferation and matrix production through affecting the expression of receptors of PDGF, which initiate both stimulatory and inhibitory signals. PMID- 9858637 TI - Exchange of Ser-4 for Val, Leu or Asn in the sequon Asn-Ala-Ser does not prevent N-glycosylation of the cell surface glycoprotein from Halobacterium halobium. AB - The archaeon Halobacterium halobium expresses a cell surface glycoprotein (CSG) with a repeating pentasaccharide unit N-glycosidically linked via N acetylgalactosamine to Asn-2 of the polypeptide (GalNAc(1-N)Asn linkage type). This aspar-agine of the linkage unit is located within the N-terminal sequence Ala-Asn-Ala-Ser-, in accordance with the tripeptide consensus sequence Asn-Xaa Ser/Thr typical for nearly every N-glycosylation site known so far, which are of the GlcNAc(1-N)-Asn linkage type. By a gene replacement method csg mutants were created which replace the serine residue of the consensus sequence by valine, leucine, and asparagine. Unexpectedly, this elimination of the consensus sequence did not prevent N-glycosylation. All respective mutant cell surface glycoproteins were N-glycosylated at Asn-2 with the same N-glycan chain as the wild type CSG. Asn-479 is N-glyco-sylated via a Glc(1-N)Asn linkage type in the wild type CSG. Replacement of Ser-481 in the sequence Asn-Ser-Ser for valine prevented glycosylation of Asn-479. From these results we postulate the existence of two different N-glycosyltransferases in H.halobium, one of which does not use the typical consensus sequence Asn-Xaa-Ser/Thr necessary for all other N glycosyltransferases described so far. PMID- 9858638 TI - Genomic organization of the murine polysialyltransferase gene ST8SiaIV (PST-1). AB - Polysialic acid (PSA) is an important regulator of cellular interactions. Two enzymes (ST8SiaII and ST8SiaIV) are capable of synthesizing PSA. In the present study, the gene encoding the murine ST8SiaIV (PST-1) has been isolated and characterized. In contrast to the ST8SiaII (STX) gene which contains six exons and spans about 80 kb, the ST8SiaIV gene comprises only five exons spanning over at least 55 kb. However, alignment of the two genes revealed that exon-intron boundaries of exons 2-5 of ST8SiaIV and exons 3-6 of ST8SiaII are located at identical sites. Differences are restricted to the 5'-region encoded by one exon in the case of ST8SiaIV, whereas the corresponding region of ST8SiaII is interrupted by a very long intron. 5'-RACE analysis of the ST8SiaIV transcript using mRNA from AtT20 cells identified two transcription start sites at positions -324 and -204 relative to the translation start codon. The promoter region of ST8SiaIV lacks TATA- and CAAT-like sequences and is enriched in G+C (60%). The promoter contains putative Sp1, AP-1, AP-2, and PEA3 binding sites, as well as a purine- and a pyrimidine-rich region. Luciferase reporter gene assays demonstrated that the region between nucleotides -443 and -162 is sufficient to direct gene expression. The induction of luciferase activity was 30- and 10-fold in the PSA-positive AtT20 and CHO cells, but only 5- and 7-fold in the PSA negative NIH-3T3 cells and in a PSA-negative subline of AtT20. Thus, although decreased in activity in PSA-negative cell lines, the basal promoter is not sufficient for the strong cell-type and tissue specific regulation of the ST8SiaIV gene, suggesting regulatory elements in the more upstream 5'-region. PMID- 9858639 TI - Recycling cell surface glycoproteins undergo limited oligosaccharide reprocessing in LEC1 mutant Chinese hamster ovary cells. AB - The ability of particular cell surface glycoproteins to recycle and become exposed to individual Golgi enzymes has been demonstrated. This study was designed to determine whether endocytic trafficking includes significant reentry into the overall oligosaccharide processing pathway. The Lec1 mutant of Chinese hamster ovary (CHO) cells lack N -acetylglucosaminyltransferase I (GlcNAc-TI) activity resulting in surface expression of incompletely processed Man5GlcNAc2 N linked oligosaccharides. An oligosaccharide tracer was created by exoglycosylation of cell surface glycoproteins with purified porcine GlcNAc-TI and UDP-[3H]GlcNAc. Upon reculturing, all cell surface glycoproteins that acquired [3H]GlcNAc were acted upon by intracellular mannosidase II, the next enzyme in the Golgi processing pathway of complex N -linked oligosaccharides (t1/2= 3-4 h). That all radiolabeled cell surface glycoproteins were included in this endocytic pathway indicates a common intracellular compartment into which endocytosed cell surface glycoproteins return. Significantly, no evidence was found for continued oligosaccharide processing consistent with transit through the latter cisternae of the Golgi apparatus. These data indicate that, although recycling plasma membrane glycoproteins can be reexposed to individual Golgi derived enzymes, significant reentry into the overall contiguous processing pathway is not evident. PMID- 9858640 TI - Cloning, expression, purification, and characterization of the acid alpha mannosidase from Trypanosoma cruzi. AB - The acid alpha-mannosidase of Trypanosoma cruzi is a broad-specificity hydrolase involved in the catabolism of glycoconjugates, presumably in the digestive vacuole. We have cloned the alpha-mannosidase gene from a T.cruzi epimastigote genomic library. The alpha-mannosidase gene was determined to be single copy by Southern analysis, and similar sequences were not detected in genomic digests of either Trypanosoma brucei or Leishmania donovani. The coding region was subcloned into the Pichia pastoris expression vector pPICZ, and alpha-mannosidase activity was detected in the medium of induced cultures. The recombinant alpha-mannosidase demonstrated a pH optimum, inhibition by swainsonine, Km, and substrate specificity consistent with the characteristics of the alpha-mannosidase previously purified from T.cruzi epimastigotes. The recombinant enzyme was purified 103-fold from the culture medium of Pichia pastoris and had a native molecular mass of 359 kDa by gel filtration. A combination of SDS-PAGE, deglycosylation with endo H, and NH2-terminal sequencing indicates that the enzyme is originally synthesized as a homodimeric polypeptide that is subsequently cleaved to form a heterotetramer composed of 57 and 46 kDa subunits. A polyclonal antibody raised to the recombinant enzyme was shown to immunoprecipitate the alpha-mannosidase from T.cruzi cell extracts and will be used in future immunolocalization studies. PMID- 9858641 TI - Transbilayer movement of Glc-P-dolichol and its function as a glucosyl donor: protein-mediated transport of a water-soluble analog into sealed ER vesicles from pig brain. AB - The results described in the accompanying article support the model in which glucosylphosphoryldolichol (Glc-P-Dol) is synthesized on the cytoplasmic face of the ER, and functions as a glucosyl donor for three Glc-P-Dol:Glc0-2Man9-GlcNAc2 P-P-Dol glucosyltransferases (GlcTases) in the lumenal compartment. In this study, the enzymatic synthesis and structural characterization by NMR and electrospray-ionization tandem mass spectrometry of a series of water-soluble beta-Glc-P-Dol analogs containing 2-4 isoprene units with either the cis - or trans -stereoconfiguration in the beta-position are described. The water-soluble analogs were (1) used to examine the stereospecificity of the Glc-P-Dol:Glc0 2Man9GlcNAc2-P-P-Dol glucosyltransferases (GlcTases) and (2) tested as potential substrates for a membrane protein(s) mediating the transbilayer movement of Glc-P Dol in sealed ER vesicles from rat liver and pig brain. The Glc-P-Dol-mediated GlcTases in pig brain microsomes utilized [3H]Glc-labeled Glc-P-Dol10, Glc-P (omega, c )Dol15, Glc-P(omega, t,t )Dol20, and Glc-P-(omega, t,c )Dol20as glucosyl donors with [3H]Glc3Man9GlcNAc2-P-P-Dol the major product labeled in vitro. A preference was exhibited for C15-20 substrates containing an internal cis -isoprene unit in the beta-position. In addition, the water-soluble analog, Glc-P-Dol10, was shown to enter the lumenal compartment of sealed microsomal vesicles from rat liver and pig brain via a protein-mediated transport system enriched in the ER. The properties of the ER transport system have been characterized. Glc-P-Dol10was not transported into or adsorbed by synthetic PC liposomes or bovine erythrocytes. The results of these studies indicate that (1) the internal cis -isoprene units are important for the utilization of Glc-P-Dol as a glucosyl donor and (2) the transport of the water-soluble analog may provide an experimental approach to assay the hypothetical "flippase" proposed to mediate the transbilayer movement of Glc-P-Dol from the cytoplasmic face of the ER to the lumenal monolayer. PMID- 9858642 TI - Topological studies on the enzymes catalyzing the biosynthesis of Glc-P-dolichol and the triglucosyl cap of Glc3Man9GlcNAc2-P-P-dolichol in microsomal vesicles from pig brain: use of the processing glucosidases I/II as latency markers. AB - In the current model for Glc3Man9GlcNAc2-P-P-Dol assembly, Man5GlcNAc2-P-P-Dol, Man-P-Dol, and Glc-P-Dol are synthesized on the cytoplasmic face of the ER and diffuse transversely to the lumenal leaflet where the synthesis of the lipid bound precursor oligosaccharide is completed. To establish the topological sites of Glc-P-Dol synthesis and the lipid-mediated glucosyltransfer reactions involved in Glc3Man9GlcNAc2-P-P-Dol synthesis in ER vesicles from pig brain, the trypsin sensitivity of Glc-P-Dol synthase activity and the Glc-P-Dol:Glc0-2Man9GlcNAc2-P P-Dol glucosyltransferases (GlcTases) was examined in sealed microsomal vesicles. Since ER vesicles from brain do not contain glucose 6-phosphate (Glc 6-P) phosphatase activity, the latency of the lumenally oriented, processing glucosidase I/II activities was used to assess the intactness of the vesicle preparations. Comparative enzymatic studies with sealed ER vesicles from brain and kidney, a tissue that contains Glc 6-P phosphatase, demonstrate the reliability of using the processing glucosidase activities as latency markers for topological studies with microsomal vesicles from non-gluconeogenic tissues lacking Glc 6-P phosphatase. The results obtained from the trypsin-sensitivity assays with sealed microsomal vesicles from brain are consistent with a topological model in which Glc-P-Dol is synthesized on the cytoplasmic face of the ER, and subsequently utilized by the three Glc-P-Dol-mediated GlcTases after "flip-flopping" to the lumenal monolayer. PMID- 9858643 TI - The effect on IgG glycosylation of altering beta1, 4-galactosyltransferase-1 activity in B cells. AB - An absence of galactose on the N-linked oligosaccharides of immunoglobulin G (IgG) has been shown to affect the functional activity of the antibody molecule. In patients with rheumatoid arthritis there is an increased proportion of IgG which lacks galactose and correspondingly lower levels of beta1, 4 galactosyltransferase (beta4Gal-T) activity. The recent demonstration of several expressed beta4Gal-T genes in man raises the possibility that the enzyme responsible for the decreased IgG galactose is not the "classical" beta4Gal-T (beta4Gal-T1). To directly address the question of whether reduced beta4Gal-T1 would lead to reduced IgG galactose, the level of beta4Gal-T1 in a human IgG secreting B cell line was specifically altered using stable transfection with sense (SpcDNA3-Gal-T1) or antisense (ASpcDNA3-Gal-T1) human beta4Gal-T1 cDNA. SpcDNA3-Gal-T1 B cell transfectants expressed up to a 2.5-fold higher level of beta4Gal-T enzyme activity for the exogenous neoglycoconjugate acceptor GlcNAc pITC-BSA than did ASpcDNA3-Gal-T1 transfectants. Flow cytometric analysis with Ricinus communis agglutinin I (RCAI) revealed an overall greater number of Galbeta1,4GlcNAc structures in the fixed and permeabilized SpcDNA3-Gal-T1 B cell transfectants compared with the ASpcDNA3-Gal-T1 transfectants. Moreover, there was increased galactosylation of IgG secreted from the SpcDNA3-Gal-T1 transfectants relative to the ASpcDNA3-Gal-T1 B cell transfectants. Alteration of the level of the "classical" beta4Gal-T (beta4Gal-T1) in B cells therefore affects IgG glycosylation. PMID- 9858644 TI - Anti-GM3 antibodies activate calcium inflow and inhibit platelet-derived growth factor beta receptors (PDGFbetar) in T51B rat liver epithelial cells. AB - Glycolipids expressed in the plasma membrane regulate a variety of cellular processes including intracellular calcium dynamics. We used flow cytometry to characterize the glycoconjugates on the plasma membrane of T51B liver epithelial cells. Antibodies against glycolipids found to be present were tested for their ability elevate intracellular calcium. An antibody against GM3 (DH2) nearly doubles intracellular calcium while an antibody against type II chains (1B2) increases calcium to nearly four times the baseline level, similar to levels obtained with epidermal growth factor (EGF). The antibodies stimulated calcium inflow but did not trigger calcium release from internal stores. In addition DH2 but not 1B2 inhibited platelet-derived growth factor beta receptor (PDGFbetar) function. This is the first demonstration of activation of calcium inflow by agents that bind GM3 and type II chains. The ganglioside-mediated calcium inflow is likely to stimulate secretion by these liver cells. PMID- 9858646 TI - Monoclonal antibody 91.9H raised against sulfated mucins is specific for the 3' sulfated Lewisa tetrasaccharide sequence. AB - The IgG1hybridoma antibody, 91.9H, was originally raised against sulfated mucins isolated from normal human colonic mucosa. Previous studies have shown that the 91.9H antigen is expressed on normal colonic epithelial cells and the sulfomucins that they produce, but not in the normal small intestine and stomach. Tissue specific changes occur in 91.9H antigen expression in disease: the antigen diminishes in colonic carcinomas, whereas in regions of gastric mucosa showing intestinal metaplasia and in gastric carcinomas, the antigen is expressed as a "neo-antigen." This report is concerned with elucidation, by the neoglycolipid technology, of the determinant recognized by antibody 91.9H using sulfated and sialyl oligosaccharides of Lewisa(Lea) and Lextypes, and analogs that lack sulfate, sialic acid, or fucose. Binding experiments with the lipid-linked oligosaccharides immobilized on chromatograms or on microwells, and inhibition of binding experiments with free oligosaccharides based on di-, tri- and tetrasaccharide backbones, show that the 91.9H antigenic determinant is based on a trisaccharide backbone, and consists of the 3'-sulfated Leatetrasaccharide sequence, which is a potent ligand for the E- and L-selectins. The antibody gives a relatively low signal with the 3'-sulfated non-fucosylated backbone, and has no detectable cross-reaction with the 3'-sulfated Lexisomer, nor with sialyl-Leaand Lexanalogues. Antibody 91.9H is a valuable addition, therefore, to the repertoire of reagents for mapping details of the distribution, and determining the relative importance of sulfated and sialyl oligosaccharides as ligands for the selectins, in normal and pathological epithelia and endothelia. PMID- 9858645 TI - Neurotrophic growth factors stimulate glycosaminoglycan synthesis in identified retinal cell populations in vitro. AB - Glycosaminoglycans (GAG) are known to participate in central nervous system processes such as development, cell migration, and neurite outgrowth, but little is known with respect to their regulation through soluble neurotrophic factors. In the present study, we have addressed this issue using cell culture models of three distinct cell populations derived from young rat retinas, namely, purified M uller glia, pigmented epithelium, and neurons respectively. Cultures were maintained in chemically defined media in the presence or absence of either basic fibroblast or epidermal growth factor. In control glial and epithelial cultures, hyaluronic acid dominated the soluble GAG pool, with lesser contributions from dermatan sulfate, chondroitin sulfate, and heparan sulfate (in decreasing order). Retinal neuronal GAG were almost exclusively chondroitin sulfate (approximately 90%). Treatment of glial and epithelial cultures with either factor led to dose dependent increases in especially hyaluronic acid synthesis (a maximum 6-fold increase relative to control levels), with smaller but consistent changes in chondroitin sulfate. Similar treatment of retinal neurons did not lead to any changes in GAG synthesis. These data indicate that glia and pigment epithelia are the principal sources of GAG components in retina at least in vitro, and that endogenous neurotrophic growth factors can greatly modify GAG synthesis in these two retinal cell populations. Such data suggest that a delicate balance may exist between growth factor availability and glycoconjugate metabolism in vivo, participating in normal or pathological states of the retina. PMID- 9858647 TI - Salt and water transport across the alveolar epithelium in the developing lung: correlations between function and recent molecular biology advances (Review). AB - Significant progress have been made in understanding the mechanisms of alveolar fluid clearance at the time of birth and the transition from placental oxygenation to air breathing. During fetal life, the mammalian lung is a fluid filled secretory organ that fills no respiratory function. Its potential air spaces are filled with fluid that is actively secreted in response to an osmotic force generated by Cl(-)-secretion and the fluid-filled lung is necessary for a proper development of the air-breathing lung. As term approaches, net Cl(-) secretion decreases, which is accompanied by a decreased secretion rate of the fluid into the air spaces. Concomitantly with the decreased Cl(-)-secretion, the alveolar epithelium begins to absorb Na+ to prepare for fluid absorption and the air breathing life. The causes for the decreased Cl(-)-secretion and the beginning of the Na+ absorption are not clear. Alterations in the hormonal milieu of the lung as well as changes in plasma stress hormone levels have been suggested to play roles. The switch from a placental oxygenation to pulmonary oxygenation requires that the fluid in the air spaces be rapidly removed from the lung lumen. Recent studies have demonstrated that removal of the alveolar fluid at birth is regulated via endogenous plasma epinephrine in the newborn. Molecular, cellular, and whole animal in vivo studies have demonstrated that fluid absorption at birth is related to expression and function of the epithelial sodium channel (ENaC). Several different in vivo and in vitro preparations have been used to investigate the mechanisms of alveolar fluid transport, primarily in adult lungs and have demonstrated that alveolar fluid absorption is driven by active Na+ transport. Both catecholamine-dependent and -independent regulatory mechanisms have been identified, probably acting on ENaC and other apical sodium channels and/or the basolaterally located Na+, K(+)-ATPase. Future studies are needed to integrate new insights to the molecular mechanisms behind fluid clearance with their function in both normal and pathological lungs. PMID- 9858648 TI - Role of tetrahydrobiopterin in the function of nitric oxide synthase, and its cytoprotective effect (Review). AB - This review discusses the role of (6R)-5,6,7,8-tetrahydrobiopterin (H4 biopterin) in the function of nitric oxide synthase (NOS), and the protective effect of H4 biopterin against nitric oxide (NO)- and/or reactive oxygen species-induced cytotoxicity. Although NOS releases NO, which regulates vascular tone and immune surveillance under normal conditions, NOS seems to produce superoxide anion and hydrogen peroxide when H4 biopterin, one of the cofactors, or L-arginine, a substrate, is decreased, suggesting the possibility that NOS is a source of reactive oxygen species under pathological conditions. Moreover, simultaneous release of NO and reactive oxygen species in the presence of suboptimal concentrations of H4 biopterin and/or L-arginine may be highly toxic, since NO reacts with superoxide anion and hydrogen peroxide to form peroxynitrite, singlet oxygen and the hydroxyl radical, which are toxic. An increase in H4 biopterin content in cells obviates NOS dysfunction (production of reactive oxygen species instead of NO) and protects the cells against NOS dysfunction-related cell injury. Moreover, H4 biopterin has a strong scavenging activity for reactive oxygen species, and inhibits thier cytotoxicity. H4 biopterin is also likely to reduce NO-induced cytotoxicity. Thus, H4 biopterin is not only an important regulator of NOS function, but is also an intracellular antioxidant. NO and reactive oxygen species are known to be implicated in the development of many pathological states. It is possible that H4 biopterin could be effective for treating many diseases, such as ischemia-reperfusion injury, inflammation and diabetes mellitus. PMID- 9858649 TI - The non-env tropism of HIV/SIV (Review). AB - The tropism of human and simian immunodeficiency viruses (HIV and SIV) determined by sequences other than env has been studied. The restriction of HIV-1 replication in monkey cells was demonstrated to be regulated by viral non-env sequence. Likewise, the gag-pol region of SIVagm (virus of the African green monkey) genome was found to be responsible for growth restriction in human cells of the virus. No viral DNA synthesis was detected in cells nonpermissive for the viruses. In addition, a number of HIV-1 gag gene mutants, which have an early defect in viral replication cycle and direct no viral DNA synthesis in some cells, exhibited a phenotype of host range mutant. Taken together, it can be concluded that the viral tropism associated with the uncoating/ reverse transcription process does exist in HIV/SIV replication. Furthermore, many of the accessory gene mutants of HIV/SIV exhibit host cell-dependent replication property. In this review, we summarize these examples of non-env tropism of HIV/SIV. PMID- 9858650 TI - Setting up of an original computer-assisted methodology to characterize in vitro drug-induced anti-angiogenic effects. AB - The development of angiogenesis within a tumor brings on a sequence of extremely complex molecular events. We have developed a methodology which enables a wide set of biological parameters to be quantitatively determined in the field of anti angiogenesis pharmacology. This methodology which includes a video cell tracking device, is unique because it offers the possibility of evaluating the specific influence of a given compound with potential anti-angiogenic properties on cell cycle kinetics, cell death, global cell line growth, and cell motility. We chose TNP-470, a synthetic analogue of fumagilin, to test our methodology on HUVEC cell lines taken from various human umbilical cord veins. The experiments carried out with TNP-470 did not confirm all the data reported in the literature. Our results show that i) TNP-470 could be considered as a cytotoxic agent; ii) this compound had an apparently marginal cytostatic effect; and iii) it did not increase the apoptosis level. Our methodology also revealed that the HUVEC cell lines are very heterogeneous in terms of different biological parameters. This highlights the problem of the reproductibility of the result. PMID- 9858651 TI - CGH-detected DNA sequence copy number amplifications can be confirmed by interphase-FISH: new possiblities for prognostic approaches in oral squamous cell carcinomas. AB - In order to control the data obtained by comparative genomic hybridization (CGH) on DNA sequence copy number amplifications, 20 oral squamous cell carcinomas (SCC) were subjected to interphase fluorescence in situ hybridization (I-FISH) examination using specific DNA probes for the oncogenes int2 and erbB-2, and the corresponding centromeric probes of chromosomes 11 and 17. In all cases characterized by distinct peaks of the CGH profile on the critical chromosomal segment 11q13, these data could be clearly substantiated by the I-FISH analyses using the int2 probe and estimating the signal index, the int2/centromer 11 relation, and the fraction of nuclei with high int2 signal numbers. In addition, I-FISH detected smaller cell fractions with high signal numbers (and/or signal clusters) in some tumors which were not definitely conspicuous in CGH. In contrast to int2, erbB-2 amplification apparently does not play a major role in oral SCCs, as the blurred peaks of CGH profiles on chromosome 17ql 1.2-q12 corresponded well with the findings of I-FISH using the erbB-2 probe. Gain of a whole chromosome 17 is apparently a rather common feature of these tumors. In conclusion, the combination of interphase FISH with oncogene-specific probes and CGH is regarded as a valuable means of practical molecular cytogenetic analysis of oral SCCs which could eventually achieve high practical importance in the pathologic analysis of these tumors and in prognosis of their development. PMID- 9858652 TI - Implication of the glycoxidation and lipoxidation reactions in the pathogenesis of dialysis-related amyloidosis (Review). AB - Dialysis-related amyloidosis is recognized as a serious bone and joint complication in long-term dialysis patients. Beta2-microglobulin has been demonstrated to be a major constituent of the amyloid fibrils. However, the molecular pathogenesis of this disorder remains unknown. Recent biochemical and immunohistological studies have identified a new modification of beta2 microglobulin in the amyloid fibrils, i.e., the advanced glycation end products (AGEs). AGEs are formed by non-enzymatic glycative and oxidative (glycoxidation) reactions. The levels of AGEs, such as pentosidine and carboxymethyllysine (CML), are elevated in both the plasma proteins and skin collagen of non-diabetic dialysis patients several times more than in normal subjects. The AGE accumulation in uremia cannot be attributed to hyperglycemia, nor simply to their decreased renal clearance. Recently, gathered evidence has suggested that, in uremia, an increase in carbonyl compounds, derived from both carbohydrates and lipids, modifies proteins, leading to the augmentation of the production of not only AGEs, but also the advanced lipoxidation end products (ALEs). Uremia might thus be a state of carbonyl overload with potentially damaging proteins ('carbonyl stress'). Immunohistochemical studies, with antibodies specific to AGEs and ALEs, identified carbonyl stress in long-lived beta2-microglobulin amyloid deposits. Furthermore, proteins modified by carbonyl stress exhibit a variety of biological activities towards several types of cells, which might partially account for dialysis arthropathies. PMID- 9858653 TI - Effects of irradiation on telomerase activity in human lymphoma and myeloma cell lines. AB - The effect of high-dose irradiation on telomerase activity was examined in some human lymphoma (DL40, DL95, DL110) and myeloma (U266) cell lines. The survival rate was reduced in DL40, DL110 and U266 by irradiation. Irradiation, however, showed no effect on the rate of DL95. Telomerase activity was detected in non irradiated samples of all cell lines, as measured by PCR-based telomeric repeat amplification protocol assay. The telomerase activity increased 2-6.5 fold by irradiation. Especially in DL110, the activation increased in a dose-dependent manner. In the early phase after irradiation, we observed no correlation between telomerase activity and cell viability, suggesting that telomerase-mediated chromosome healing might not be a major cause and/or not sufficiently effective to protect the cells from irradiation. PMID- 9858654 TI - Infection with the human immunodeficiency virus type 2: epidemiology and transmission (Review). AB - Human immunodeficiency virus type 2 (HIV-2) is known to be one of the agents that causes acquired immunodeficiency syndrome (AIDS). It has been present in West Africa since the 1960s and is currently epidemic there. Compared with human immunodeficiency virus type 1 (HIV-1), HIV-2 is genomically different. Furthermore, it is less prevalent worldwide than HIV-1. In West Africa, seropositive rates of HIV-2 are higher in urban versus rural communities, however, there are no gender differences. Sexual contact and vertical transmission are known modes of infectivity, though HIV-2 is less contagious than HIV-1. PMID- 9858655 TI - Abnormal stress response and increased fighting behavior in mice lacking the bcr gene product. AB - The in vivo function of proteins which regulate activity of the GTPase Rac is largely unknown. Here we establish that mice lacking bcr, a known GTPase activating protein for Rac, exhibit a defect in the regulation of both hormonal and behavioral stress responses. Bcr null mutants demonstrate prolonged elevation of plasma glucocorticoids and increased fighting in males in response to physiological and social stress, respectively. Combined biochemical and behavioral data indicate that bcr is involved in mediating the cellular effects of glucocorticoids, specifically down-regulation of the stress-activated hippocampal hypothalamic-pituitary-adrenal axis. PMID- 9858656 TI - Molecular pharmacology of methyl-3,5-diiodo-4 (4'methoxyphenoxy) benzoate (DIME) and its non-hydrolyzible ethanone analog (DIPE) (Review). AB - A molecular structural relationship of thyroid hormones to methyl-3,5-diiodo-4 (4'-methoxy-phenoxy) benzoate (DIME) and 1-[3,5-diiodo-4-(4'-methoxyphenoxy) phenyl]-ethanone) (DIPE) and to apoptosis-mediated metamorphogenic mechanisms is postulated. DIME disrupts microtubule assembly already in anaphase, preparing cells for G2/M block, chromosome aggregation and caspase-3 mediated apoptosis. Cooperative action of DIME and vincristine, defining mutually exclusive cellular sites, identifies microtubules as primary drug targets followed by downstream cellular consequences, leading to cell death. Absence of in vivo toxicity of DIME appears to be related to impermeability to DIME of normal cells, but not of tumor cells in vivo. Normal tissue cells hydrolyze DIME but most tumor cells, except lung cancer cells, do not. DIPE, being resistant to enzymatic hydrolysis, is equally effective in all tumor cells. PMID- 9858657 TI - Poly (ADP-ribose) polymerase, a potential target for drugs: Part II. Regulation of differentiation by the poly ADP-ribose system. AB - It is shown that eukaryotic differentiation is specifically sensitive to pADPRT regulation in Trypanosomna, Leishmania and Mytilus models. There is powerful inhibition of early differentiation without cell toxicity by pADPRT ligands. PMID- 9858658 TI - Angiotensin II receptors on colorectal carcinoma cells. AB - The presence of angiotensin II receptors was found on cells of three colorectal carcinoma cell lines. The binding assays with 125I-labelled angiotensin II and ligands specific for angiotensin AT1 or AT2 receptors showed that angiotensin receptors on colorectal cancer cells are mostly of the AT2 type. The binding capacity of tumor cells was not significantly changed by butyrate-induced differentiation. PMID- 9858659 TI - Quantitative trait loci for blood glucose confirm diabetes predisposing and protective genes, Iddm4 and Iddm5r, in the spontaneously diabetic BB/OK rat. AB - Several crossing studies using diabetic BB/OK and diabetes-resistant rat strains have clearly shown that the MHC class-II-genes of the RT1u haplotype (Iddm1) and the lymphopenia (Iddm2) are essential but not sufficient for type 1 diabetes development. The search for additional diabetogenic genes revealed predisposing non-MHC genes, Iddm3 and Iddm4, and a diabetes protective gene, Iddm5r, cosegregating with diabetes in the BB/OK rat subline. These findings were based on cosegregation studies comparing allele frequencies between diabetic and non diabetic cross hybrids. Since, type 1 diabetes is characterised by hyperglycaemia we analysed 22 diabetic and 43 non-diabetic [(BB x SHR)FI x BB] backcross hybrids (28M:37F) which were already homozygous for Iddml and Iddm2 to search for quantitative trait loci (QTLs) affecting blood glucose in BB/OK rats. The QTL analysis using 117 microsatellite markers located on 19 autosomal chromosomes and the X chromosome, revealed suggestive linkage for blood glucose at the same position for diabetics (lod score 3.1) and non-diabetics at an age of 16 weeks at locus D6Mgh2 on chromosome 6 (lod score 1.9). In contrast, the peak for nondiabetics at an age of 28 weeks (lod score 3.1) was located in the region on chromosome 1 flanked by D1Mgh12 and D1Mit14, whereas the peak for diabetics (lod score 1.9) was found between Sa and Igf2. The distance between two peaks is ca. 50 cM. These findings are consistent with previously described results and provide strong evidence on the relevance of the described region for the development of diabetes not only in the rat, but, regarding the chromosomal homology also in human. PMID- 9858660 TI - ATP inhibition of proliferation of immortalized human fibroblasts is greater than that of normal human diploid fibroblasts. AB - It is known that cancers develop by a multi-step process. Normal cells are first immortalized, and then transformed into tumorigenic cells. Normal human cells are very rarely immortalized, but once they are, they are relatively easily transformed into tumorigenic cells. This indicates that the immortalization step plays a critical part in the development of human cancers. Thus, elucidation of the mechanisms of this step would shed light on the process of carcinogenesis in human cells. To understand the causes of immortalization, it is important to determine the differences in cellular phenotype between immortalized and normal human cells. In this study, we found that immortalized human fibroblasts were more sensitive to the growth inhibitory effects of ATP than normal human fibroblasts. ADP was as effective as ATP, but AMP, adenosine, and phosphoric acid were not. These results indicate that a high-energy bound of ATP and ADP may contribute to the growth inhibition of the cells. When the immortalized cells were pulse-labeled with [32P]-ATP, 30-, 31-, 33- and 40-kDa membrane fraction proteins were more prominently labeled in the immortalized cells than in the normal cells. At present, the characteristics of these proteins are being investigated. PMID- 9858661 TI - Intrauterine events and the programming of adulthood disease: the role of fetal glucocorticoid exposure (Review). AB - There is increasing epidemiological evidence in humans which associates low birth weight with later cardiovascular and metabolic disorders including hypertension, insulin resistance, hyperlipidaemia and death from ischaemic heart disease. The molecular mechanisms underlying this link are unknown but fetal glucocorticoid exposure may play a role. In adult mammals, glucocorticoid hormones are involved in control of several physiological processes that maintain homeostasis including coordination of responses to stress. During development, glucocorticoids have important regulatory functions to prepare the organism for metabolic adaptations necessary for extrauterine life. Fetal glucocorticoid load is, in part, regulated by placental and fetal 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) which catalyses a rapid breakdown of maternal and fetal glucocorticoids into inert products. Supraphysiological doses of glucocorticoids retard fetal growth, and human intrauterine growth retardation is associated with elevated cortisol levels. Recent studies have shown that exposing rats to excessive glucocorticoids in utero reduces birth weight and causes permanent hypertension and hyperglycaemia in the adult offspring. These observations show that glucocorticoids could be the link between low birth weight and later disease. Understanding of the molecular details involved in prenatal glucocorticoid action may provide novel insights into the pathogenesis of common cardiovascular and metabolic disorders. PMID- 9858662 TI - Effects of carbaryl and naphthalene on rat hepatic CYP1A1/2: potential binding to Ah receptor and 4S benzo(a)pyrene-binding protein. AB - Activation of the CYP1A1 gene has been described to be mediated by the cytosolic Ah receptor (AhR) and a possible cooperative role of the 4S benzo(a)pyrene binding protein (4S protein). Carbaryl (CAR) has been shown to induce human CYP1A1 gene expression without binding to the human AhR. In this study, Sprague Dawley rats received a single i.p. dose of 20, 80, 150 micromol/kg CAR or NAPn (naphthalene, the aromatic part of CAR) and were sacrificed after 24 h. CAR increased ethoxyresorufin-O-deethylase and methoxyresorufin-O-demethylase activities, the level of CYP1A1, 1A2 proteins, and CYP1A1 mRNA at the highest dose, whereas NAPn showed no effects. Moreover, CAR, naphthol (its major metabolite) and NAPn were not ligands in vitro of the TCDD binding site of AhR or the benzo(a)-pyrene binding site of 4S protein in rat, neither was CAR a ligand of these two binding sites in mice, dog, monkey or human. Molecular properties of CAR were evaluated and showed that this molecule is far from the structural characteristics of CYP 1A1 specific inducers although a planar conformation can be achieved with an energy < 5 kJ x mol(-1). The data demonstrated that CAR could also modulate the AhR-mediated responses, even though it did not meet the structural requirements to be ligand of AhR. PMID- 9858663 TI - Differential nitric oxide release and sensitivity to injury in different murine mammary tumor cell lines. AB - The purpose of this study was to determine whether nitric oxide (NO) production by different mammary tumor cell lines correlated with their sensitivity to NO mediated injury. Three mammary tumor cell lines LM2, LM3 and LMM3 syngeneic to BALB/c mice were cultured in vitro with IFNgamma + LPS. Different levels of NO production among the three lines were detected in culture supernatants. The only tumor cell line which did not produce NO (LM2) showed the highest sensitivity to SNP-derived NO cytotoxicity (87%), while LM3 and LMM3 which both produced higher levels of NO than LM2, showed lower cytotoxicity by SNP (39% and 22% respectively). Spleen cells (SC) from M2 tumor bearing mice (TBM) were able to lyse LM2 cells by NO-dependent mechanisms. SC from M3-TBM exerted cytotoxicity against LM3 cells mainly by NO-independent mechanisms. Thus, we postulate an inverse correlation between NO production and NO mediated cytotoxicity in the three mammary tumor cell lines. It is possible that tumor cells producing NO develop mechanisms to resist NO injury. PMID- 9858664 TI - Role of the glycosyl-phosphatidylinositol anchor in the intracellular transport of a transmembrane protein in Madin-Darby canine kidney cells. AB - In order to compare the trafficking of proteins with different membrane anchors, we have constructed and expressed three different recombinant forms of neutral endopeptidase (NEP) in MDCK cells. The wild type form of NEP (WT-NEP) is attached to the plasma membrane by a single N-terminal membrane spanning domain, whereas the glycosylphosphatidylinositol-anchored form of the protein (GPI-NEP) contains a C-terminal GPI anchor. A double anchored form of NEP (DA-NEP) was also constructed, that contains both the original N-terminal membrane spanning domain and a C-terminal GPI anchor. We show here that WT-NEP, GPI-NEP and DA-NEP, which are all apically targeted in MDCK cells, behave differently when subjected to Triton X-100 solubilisation: despite the presence of the transmembrane anchor DA NEP behaves as a GPI-anchored protein. This suggests that the GPI anchor of DA NEP is dominant over the transmembrane anchor of the native protein to determine its pattern of solubility in Triton X-100. PMID- 9858666 TI - Role of fibroblast growth factor during early midbrain development in Xenopus. AB - Genes encoding fibroblast growth factors (FGFs) are expressed in early Xenopus neurulae in the prospective midbrain-hindbrain boundary (MHB) region of the neural plate. These expression domains overlap those of XWnt-1 and XEn-2, raising the question of the role of FGF signalling in the regulation of these genes, and more generally about the function of FGF during Xenopus midbrain development. We report that explants from the prospective MHB grafted into the anterior neural plate in midneurula stage embryos induce XWnt-1 expression and, at a lower frequency, XEn-2 expression in the vicinity of the graft. Such a process is likely to involve FGF signalling. Implantation of FGF4- or FGF8-soaked beads in the prospective forebrain at neurula and tailbud stages causes the up-regulation of XWnt-1 and XEn-2 in the dorsal and lateral region of the anterior midbrain. This effect is not relayed by endogenous FGF genes since exogenous FGFs inhibit the expression of endogenous XFGF3 or XFGF8. However, consequences of grafting MHB or implanting FGF4 or FGF8 beads on tadpole brain development are different. MHB grafts induce ectopic mesencephalic structures, strongly suggesting that a region homologous to the isthmic organizer of amniotes is specified as early as the midneurula stage. In contrast, exogenous FGFs do not cause the formation of ectopic mesencephalic structures but an overgrowth of mesencephalon and diencephalon. We propose that FGF signals from the prospective MHB play a crucial role in the spatial regulation of XWnt-1 and XEn-2 expression in the posterior midbrain, but that the full organizing activity of the MHB involves other factors in combination with FGF. PMID- 9858665 TI - Cytochrome P450 1A1 in rat peripheral blood lymphocytes: inducibility in vivo and bioactivation of benzo[a]pyrene in the Salmonella typhimurium mutagenicity assay in vitro. AB - The presence and inducibility of CYP1A1 in freshly isolated peripheral blood lymphocytes was examined in untreated rats and in rats pretreated with agents known to induce the enzyme in other tissues, as well as dexamethasone [CAS #50-02 2], which is not commonly associated with CYP1A1 induction. CYP1A1 but not CYP1A2 was detected by Western blot analysis of lymphocytes from untreated rats and was induced in lymphocytes from rats treated with the known CYP1A inducers beta naphthoflavone [CAS #6051-87-2] or 3-methylcholanthrene [CAS #56-49-5] (7.3 fold), cigarette smoke (2. 8-fold), and pyridine [CAS #108-86-1] (2.6-fold). CYP1A1 was also induced in lymphocytes from rats treated with the nonprototypic inducer dexamethasone (17.7-fold) or bromobenzene [CAS #108-86-1] (3. 9-fold). Lymphocyte homogenate from rats treated with the inducers also catalyzed NADPH dependent bioactivation of benzo[a]pyrene [CAS #50-32-8] to mutagens. The benzo(a)pyrene mutagenicity was detected using Salmonella typhimurium TA100 in the Ames test, and correlated positively with lymphocyte CYP1A1 content. The data show that CYP1A1 is present in rat peripheral blood lymphocytes in vivo, and is inducible by prototypic, as well as nonprototypic, inducers of the enzyme. PMID- 9858667 TI - Recombinogenic targeting: a new approach to genomic analysis--a review. AB - Currently, recombinational cloning procedures based upon methods developed for yeast, Saccharomyces cerevisiae, are being exploited for targeted cloning and in vivo modification of genomic clones. In this review, we will discuss the development of large-insert vectors, homologous recombination-based techniques for cloning and modification, and their application towards functional analysis of genes using transgenic mouse model systems. PMID- 9858668 TI - An investigation into the lipid interactions of peptides corresponding to the C terminal anchoring domains of Escherichia coli penicillin-binding proteins 4, 5 and 6. AB - The Escherichia coli low molecular mass penicillin-binding proteins PBP4, PBP5 and PBP6 are DD-peptidases involved in murein biosynthesis. It has been suggested that these proteins may be anchored to the periplasmic face of the inner membrane via their C termini. Here, peptide homologues (P4, P5 and P6) of the PBP4, PBP5 and PBP5 C-terminal regions have been used to investigate potential protein-lipid interactions involved in this anchoring mechanism. Surface pressure changes observed for the interactions of P5 and P6 with a range of monolayers indicated that the peptides are membrane interactive and that the interactions proceeded via predominantly hydrophobic forces with only minor requirements for anionic lipid. In contrast, P4 interactions with monolayers appeared to proceed via predominantly electrostatic forces with a major requirement for anionic lipid. The lipid interactions of all three peptides were generally enhanced by low pH and for P5 and P6 were in the range of 10-15 mN m-1 whereas for P4 interactions they were in the range of 3-7 mN m-1. CD analysis implied the presence of alpha helical structure in P5 and P6 and molecular area determinations implied that P4 may also possess helical architecture in the presence of dioleoylphosphatidylglycerol monolayers. Overall, our results support the view that C-terminal amphiphilic alpha-helices are involved in the membrane anchoring of PBP5 and PBP6 and suggest that a similar mechanism could contribute to PBP4 membrane anchoring. Furthermore, we have speculated that the presence of cationic residues in the hydrophilic face of these alpha-helices may help facilitate membrane interaction. PMID- 9858669 TI - Analysis of correlated data in human biomonitoring studies. The case of high sister chromatid exchange frequency cells. AB - Sister chromatid exchange (SCE) analysis in peripheral blood lymphocytes is a well established technique that aims to evaluate human exposure to toxic agents. The individual mean value of SCE per cell had been the only recommended index to measure the extent of this cytogenetic damage until the early 1980's, when the concept of high frequency cells (HFC) was introduced to increase the sensitivity of the assay. All statistical analyses proposed thus far to handle these data are based on measures which refer to the individual mean values and not to the single cell. Although this approach allows the use of simple statistical methods, part of the information provided by the distribution of SCE per single cell within the individual is lost. Using the appropriate methods developed for the analysis of correlated data, it is possible to exploit all the available information. In particular, the use of random-effects models seems to be very promising for the analysis of clustered binary data such as HFC. Logistic normal random-effects models, which allow modelling of the correlation among cells within individuals, have been applied to data from a large study population to highlight the advantages of using this methodology in human biomonitoring studies. The inclusion of random-effects terms in a regression model could explain a significant amount of variability, and accordingly change point and/or interval estimates of the corresponding coefficients. Examples of coefficients that change across different regression models and their interpretation are discussed in detail. One model that seems particularly appropriate is the random intercepts and random slopes model. PMID- 9858670 TI - Modulation of Hedgehog target gene expression by the Fused serine-threonine kinase in wing imaginal discs. AB - The Fused (Fu) serine-threonine kinase and the Suppressor of fused (Su(fu)) product are part of the Hedgehog (Hh) signaling pathway both in embryos and in imaginal discs. In wing imaginal discs, the Hh signal induces Cubitus interruptus (Ci) accumulation and activates patched (ptc) and decapentaplegic (dpp) expression along the anterior/posterior (A/P) boundary. In this paper, we have examined the role of the Fu and Su(fu) proteins in the regulation of Hh target gene expression in wing imaginal discs, by using different classes of fu alleles and an amorphic Su(fu) mutation. We show that, at the A/P boundary, Fu kinase activity is involved in the maintenance of high ptc expression and in the induction of late anterior engrailed (en) expression. These combined effects can account for the modulation of Ci accumulation and for the precise localization of the Dpp morphogen stripe. In contrast, in more anterior cells which do not receive Hh signal, we show that Fu plays a role independent of its kinase function in the regulation of Ci accumulation. In these cells, Fu may be involved in the stabilization of a large protein complex which is probably responsible for the regulation of Ci cleavage and/or targeting to nucleus. We propose that the Fused function is necessary for the activation of full-length Ci and counteracts the negative Su(fu) effect on the pathway, leading to en, ptc and dpp expression. PMID- 9858672 TI - Evaluation of 8-oxodeoxyguanosine, typical oxidative DNA damage, in lymphocytes of ozone-treated arteriosclerotic patients. AB - In the present study we measured the amount of 8-oxo-2'-deoxyguanosine (8-oxo-dG) in DNA isolated from lymphocytes of arteriosclerotic patients undergoing ozonetherapy. Treatment of the patients with therapeutic concentration of ozone caused a significant increase over the control value in the amount of 8-oxo-dG of DNA isolated from their lymphocytes. However, only three out of six patients examined responded positively to the treatment in terms of the base damage. The increases varied among patients, and were in the range of 100-450%. This interindividual difference may at least be partly explained by recently demonstrated heritable susceptibility to ozone. PMID- 9858671 TI - Fluorescent differential display analysis of gene expression in apoptotic neuroblastoma cells. AB - Identification of differentially expressed genes will provide leads in the elucidation of the molecular mechanisms underlying neuronal cell death associated with neurodegenerative disorders. Using a high-throughput fluorescent differential display (FDD) system based on an automated DNA sequencer, we analyzed global patterns of gene expression during the apoptosis of neuroblastoma SH-SY5Y cells induced by a neurotoxin, colchicine. Initial screening of approximately 24000 cDNA bands displayed with 320 primer combinations has revealed 263 fragments showing differential expression patterns, suggesting that approximately 1% of transcripts are modulated in their expression level. Of these differentially displayed bands, we cloned 18 fragments composed of 17 distinct species and confirmed differential expression of each species by reverse transcription-PCR or Northern blot hybridization, thereby proving the reliability of the approach. These include eight derived from seven known genes, five homologous to expressed sequence tags (ESTs), and five totally lacking any homology to those deposited in the database. Among these, a novel transcript SAI1 induced prominently was characterized further and revealed to encode a putative RNA-binding protein NAPOR (neuroblastoma apoptosis-related RNA-binding protein), containing three copies of evolutionarily conserved RNA recognition motif. Since several RNA-binding proteins have been known to play crucial roles in other apoptosis systems, it is conceivable that NAPOR is also involved in the process of neuronal cell death. PMID- 9858673 TI - Doxorubicin physical state in solution and inside liposomes loaded via a pH gradient. AB - We have examined doxorubicin's (DOX) physical state in solution and inside EPC/cholesterol liposomes that were loaded via a transmembrane pH gradient. Using cryogenic electron microscopy (cryo-EM) we noted that DOX loaded to 200-300 mM internal concentrations in citrate containing liposomes formed linear, curved, and circular bundles of fibers with no significant interaction/perturbation of the vesicle membrane. The individual DOX fibers are putatively comprised of stacked DOX molecules. From end-on views of bundles of fibers it appeared that they are aligned longitudinally in a hexagonal array with a separation between fibers of approx. 3-3.5 nm. Two distinct small angle X-ray diffraction patterns (oblique and simple hexagonal) were observed for DOX-citrate fiber aggregates that had been concentrated from solution at either pH 4 or 5. The doxorubicin fibers were also present in citrate liposomes loaded with only one-tenth the amount of doxorubicin used above (approx. 20 mM internal DOX concentration) indicating that the threshold concentration at which these structures form is relatively low. In fact, from cryo-EM and circular dichroism spectra, we estimate that the DOX-citrate fiber bundles can account for the vast majority (>99%) of DOX loaded via a pH gradient into citrate buffered liposomes. DOX loaded into liposomes containing lactobionic acid (LBA), a monoanionic buffer to control the internal pH, remained disaggregated at internal DOX concentrations of approx. 20 mM but formed uncondensed fibers (no bundles) when the internal DOX concentration was approx. 200 mM. This finding suggests that in the citrate containing liposomes the citrate multianion electrostatically bridged adjacent fibers to form the observed bundles. 13C-NMR measurements of [1,5-13C]citrate inside liposomes suggested that citrate 'bound' to the DOX complex and 'free' citrate rapidly exchange indicating that the citrate-DOX interaction is quite dynamic. DOX release into buffer was relatively slow (<4% at 1 h) from liposomes containing DOX fibers (in citrate loaded to a low or high DOX concentration or in LBA liposomes loaded to a high internal DOX concentration). LBA containing liposomes loaded with disaggregated DOX, where the internal DOX concentration was only approx. 20 mM, experienced an osmotic stress induced vesicle rupture with as much as 18% DOX leakage in less than 10 min. The possible implications for this in vivo are discussed. PMID- 9858674 TI - Advantage of the use of human liver S9 in the Ames test. AB - The mutagenicity of 13 chemicals was compared using human liver S9 or liver S9 prepared from male Sprague-Dawley rats either non-treated (R-n) or pretreated with phenobarbital/5,6-benzoflavone (R-i). The test compounds used in this study were well recognized procarcinogens requiring cytochrome P450 for metabolic activation. These included polycyclic aromatic hydrocarbons, aromatic amines, heterocyclic aromatic amines, nitrosoamines, and nitropyrene. We used four human liver S9 fractions, one of which was prepared from the liver sample having higher levels of the P450-catalyzed drug metabolizing enzyme activities, a possible explanation for which was enzyme induction by anti-asthma agents for 10 years. The results of the present study are as follows: (1) there were individual differences in the magnitude of the mutagenic activity of the procarcinogens by each S9 fraction used, (2) equivalent mutagenicity of chemicals was seen with three human S9 fractions (H3, H8, and H12), while a human H14 S9 fraction showed higher P450 enzyme activity, leading to much higher mutagenicity than the other three human S9 specimens, (3) the order of magnitude of the mutagenicity of the procarcinogens using human and rat liver S9 fractions was R-i>/=H14>/=R-n>/=H3, H8, and H12, while with 2-aminoanthrathene, N-nitrosodimethylamine, and 1 nitropyrene, this relationship was H3, H8, H12, and H14>/=R-n>/=R-i. The experimental data in the present study strongly suggest that the complementary use of human liver S9 fraction in the Ames test is a much more useful tool than rat S9 for evaluation of genotoxicity to humans. PMID- 9858675 TI - 'Mass-murder' of ORFs from three regions of chromosome XI from Saccharomyces cerevisiae. AB - The complete sequence of the yeast Saccharomyces cerevisiae reveals the presence of many new genes, many of which are without homologs in databases. Characterisation of these genes by novel methods includes systematic deletion followed by phenotypic analysis of mutant strains. We have developed a hierarchical strategy for such a functional analysis of genes, in which the primary phenotypic screening is performed on groups of contiguous genes which are then reinvestigated down to the single gene level. This strategy is applied to the whole chromosome XI as part of EUROFAN (the EUROpean Functional ANalysis) program, and we present here our results on a group of 22 genes from this chromosome. This sample is representative of the results that are emerging for the whole chromosome. Out of the 22 genes deleted, three were shown to be essential, and another three genes confer a mutant growth phenotype to cells when deleted. All phenotypes have been complemented. These figures are in accordance with the previously published fraction of lethal and growth-defective deletions of single genes. We have found no synthetic phenotypes resulting from a combination of deleted genes and have always been able to attribute a mutant phenotype to a single gene. PMID- 9858676 TI - Differential expression of non-muscle myosin heavy chain genes during Xenopus embryogenesis. AB - Class II non-muscle myosins are implicated in diverse biological processes such as cytokinesis, cellularization, cell shape changes and gastrulation. Two distinct non-muscle myosin heavy chain genes have been reported in all vertebrates: non-muscle myosin heavy chain-A (NMHC-A) and -B (NMHC-B). We report here the isolation of the Xenopus homolog of NMHC-A and present a comparative analysis of the developmental and spatial expression patterns of NMHC-A and the previously isolated NMHC-B to address the role of NMHCs in Xenopus development. A 7.5 kb NMHC-A mRNA is present, maternally in unfertilized eggs and throughout embryogenesis, as well as in all adult tissues examined. An additional 8.3 kb zygotic transcript for NMHC-A is also detected, but only during embryonic stages. Whole mount in situ hybridization with tailbud stage embryos shows that NMHC-A mRNA is predominantly expressed in the epidermis, whereas NMHC-B mRNA is expressed in the somites, brain, eyes and branchial arches. Interestingly, the expression of NMHC-B in developing somites is gradually restricted to the center of each somite as differentiation proceeds. DAPI nuclear staining demonstrated that NMHC-B mRNA is colocalized with the nuclei or perinuclear area. In animal cap experiments, treatment with activin A or ectopic expression of Xbra and an activated form of Xlim1 markedly up-regulates NMHC-B as well as muscle actin mRNAs and slightly down-regulates NMHC-A mRNA, consistent with NMHC-B expression in the somitic muscle and NMHC-A expression in the epidermis. PMID- 9858677 TI - DNA damage by hydroquinone in human white blood cells: analysis by alkaline single-cell gel electrophoresis. AB - The genotoxicity of hydroquinone (HQ) in human white blood cells was investigated by means of alkaline single-cell gel electrophoresis (SCGE). The exposure of purified lymphocytes to HQ (0.5-50 microg/ml) produced significant and dose related increases in DNA migration; conversely, no induction of DNA damage was observed in leukocytes after in vitro treatment of whole blood samples (100-500 microg/ml). Similar differences in DNA damage between whole blood samples and purified lymphocytes were observed after treatments with hydrogen peroxide (H2O2, 50 microM). The DNA damaging activity of HQ was significantly (p<0.001, U-test) inhibited by exogenous catalase (250 U/ml), indicating the generation of peroxides in the mechanism of genotoxicity of HQ. Parallel experiments using the standard SCGE protocol, and an acellular method entailing the lysis of cells before HQ treatment, provided fairly similar results, indicating that HQ oxidation does not require endogenous metabolism. Experiments to compare the effectiveness of HQ in the induction of single-strand breaks and alkali-labile sites in resting cells and micronuclei in cytokinesis-blocked cells indicate that despite the extensive DNA damage detected by SCGE immediately after treatment, a significant excess of micronuclei is not observed after stimulation and in vitro cultivation. These data explain the apparent discrepancy between the high DNA damaging potential of HQ in human lymphocytes, as revealed by SCGE, and the relatively low activity reported in most cytogenetic assays with HQ on the same cell type. PMID- 9858678 TI - Role of integrins in mouse eyelid development: studies in normal embryos and embryos in which there is a failure of eyelid fusion. AB - Eyelid fusion normally occurs between E15.5 and E16.5 of mouse embryonic development and results from the migration of a population of periderm-derived epithelial cells over the corneal surface. Cell migration is known to depend on extracellular matrix receptors of the integrin family and to be regulated by growth factors. We were therefore interested that a failure of eyelid fusion has been reported in mice that are homozygous null for the transforming growth factor alpha (TGF-alpha) gene and in mice (invalpha5beta1) in which a transgenic alpha5beta1 integrin under the control of the involucrin promoter is misexpressed in differentiating keratinocytes. We examined expression of the alpha2beta1, alpha3beta1, alpha5beta1 and alpha6beta4 integrins during eyelid fusion in wild type embryos and found selective upregulation of the alpha5beta1 integrin and its ligand, fibronectin, in the migrating eyelid tip cells. In TGF-alpha null embryos, the failure of eyelid fusion was correlated with a failure to upregulate the alpha5beta1 integrin and fibronectin in the tip cells. Using beta galactosidase as a reporter gene in transgenic mice, we observed specific activity of the involucrin promoter in the eyelid tip cells. In invalpha5beta1 mice the transgenic human integrin was overexpressed not only in the tip cells but throughout the eyelid epidermis. In contrast, the endogenous, murine, alpha5beta1 integrin was only weakly expressed in the tip cells. We speculate that selective and coordinated expression of the alpha5beta1 integrin and fibronectin in eyelid tip cells is required for eyelid fusion and may be under the control of growth factors that include TGF-alpha. PMID- 9858679 TI - Temperature dependence of monovalent cation fluxes in isolated rat hearts: a magnetic resonance study. AB - Ion flux studies were performed on Langendorff-perfused rat hearts using 87Rb, 7Li and 23Na NMR at 36, 20 and 10 degreesC, and at constant extracellular pH (7.40). Using 31P NMR, the intracellular pH was estimated and the high energy phosphate content monitored. Compared to 36 degreesC (k=0.044+/-0.015 min-1), our measurements showed incomplete Rb+ efflux with a dramatically (5-fold) increased rate constant, k, at 20 degreesC, k=0.238+/-0.080 min-1. 5 microM glibenclamide, a KATP-channel inhibitor, completely depressed the hypothermia-activated Rb+ efflux at this temperature (k=0.052+/-0. 018 min-1). 7Li NMR efflux studies on KCl-arrested hearts at 20 degreesC also showed an increase (3-fold) in efflux rate constant: k=0.090+/-0.003 min-1 relative to its value at 36 degreesC. At 10 degreesC, both Rb+ and Li+ showed efflux rate constants similar to those observed at 36 degreesC, k=0.071+/-0.016 min-1 and k=0.050+/-0. 005 min-1, respectively, and the washout was complete. 31P NMR at 36, 20 and 10 degreesC indicated cytosolic alkalinization at pH values of 7.05, 7.21 and 7.40, respectively. The ion transport data could be interpreted in terms of a myocyte model allowing for temperature-dependent changes in transport coefficients. The incomplete efflux of Rb+ at 20 degreesC may indicate the existence of a mitochondrial Rb+-pool with a very low Rb+ permeability for efflux. These findings correlate with previously observed membrane phase transitions in these systems. PMID- 9858680 TI - Subdivision of the Escherichia coli K-12 genome for sequencing: manipulation and DNA sequence of transposable elements introducing unique restriction sites. AB - A transposon-based method of introducing unique restriction sites was used for subdivision of the Escherichia coli genome into a contiguous series of large non overlapping segments spanning 2.5Mb. The segments, sizes ranging from 150 to 250kb, were isolated from the chromosome using the inserted restriction sites and shotgun cloned into an M13 vector for DNA sequencing. These shotgun sizes proved easily manageable, allowing the genomic sequence of E. coli to be completed more efficiently and rapidly than was possible by previously available methods. The 9bp duplication generated during transposition was used as a tag for accurate splicing of the segments; no further sequence redundancy at the junction sites was needed. The system is applicable to larger genomes even if they are not already well-characterized. We present the technology for segment sequencing, results of applying this method to E. coli, and the sequences of the transposon cassettes. PMID- 9858681 TI - The receptor tyrosine kinase gene linotte is required for neuronal pathway selection in the Drosophila mushroom bodies. AB - The linotte (lio) mutant was first isolated as a memory mutant. The lio gene encodes a putative receptor tyrosine kinase (RTK), homologous to the human protein RYK. This gene has been independently identified in a screen for embryonic nervous system axonal guidance defects and called derailed (drl). Here, we report that linotte mutants present structural brain defects in the adult central complex (CX) and mushroom bodies (MB). linotte and derailed are allelic for this phenotype, which can be rescued by a drl+ transgene. The Lio RTK is expressed preferentially in the adult CX and MB. Our results suggest that, analogous to its role within the embryonic nervous system, the Lio RTK is involved in neuronal pathway selection during adult brain development. PMID- 9858682 TI - In vivo anticlastogenic effects of L-ascorbic acid in mice. AB - The effects of l-ascorbic acid on the frequency of micronuclei induced by model mutagens, cyclophosphamide (CP), mitomycin-C (MMC) and bleomycin (BLM) hydrochloride were tested using mouse bone marrow. Three doses of ascorbic acid (AsA) viz., 10, 30 and 60 mg/kg b.w. were tested for anticlastogenic effects. The doses of positive mutagens used were, CP-50 mg/kg, MMC-4 mg/kg and BLM-20 mg/kg b.w. Bone marrow sampling was done at 24 h after the treatment. AsA was found to be effective in reducing MN frequency induced by CP and BLM. With MMC only, the highest dose showed a slight inhibitory effect. PMID- 9858683 TI - Sunlight mutagenesis: changes in mutational specificity during the irradiation of phage M13mp2. AB - We reported previously that the mutations in phage M13mp2, a single-stranded DNA phage, induced by sunlight exposure are predominated by G-to-C transversions. We have now made an unexpected observation that an exposure to sunlight for a short period of time results in induction mainly of C-to-T transitions while a longer exposure results in the induction of G-to-C transversions. This peculiar phenomenon suggests that DNA damage formed by initial sunlight exposure can be transformed during an elongated exposure. 7, 8-Dihydro-8-oxoguanine (8-oxoG) in DNA might be involved in the shift of the mutational specificity, as 8-oxoG was formed in the phage DNA upon the sunlight exposure. We also compared the mutagenic activity of UVB irradiation with that of sunlight exposure. The results demonstrate that the genotoxic properties of sunlight and UVB in phage M13mp2 mutagenesis are different. The shift in the mutational specificity associated with the dose of the sunlight may call for general cautions in the studies of agent-induced mutagenesis. PMID- 9858684 TI - Targeting and retrofitting pre-existing libraries of transposon insertions with FRT and oriV elements for in-vivo generation of large quantities of any genomic fragment. AB - A procedure is described that converts the pre-existing transposon insertion libraries to a collection of 'pop-out' strains, each allowing generation of 20- to 100-kb genomic fragments directly from the genome. The procedure consists of two steps: (1) single transposon insertions are targeted and retrofitted with excision and amplification elements (FRT and oriV), by homologous recombination with an FRT-oriV-carrying plasmid; and (2) two retrofitted neighbouring transposons are brought together by P1 transduction. From each strain, a 20- to 100-kb genomic fragment, bound by a pair of retrofitted transposons, could be excised and amplified upon supplying in trans the excision (Flp) and replication (TrfA) functions. To enhance the efficiency of crossing-in the FRT-oriV cassette, we transiently increased the copy number of our retrofitting plasmids using a temperature-sensitive TrfA-supplying helper plasmid. Using FRT-oriV and helper plasmids, we retrofitted four Tn10KmR and three Tn10CmR insertions. Subsequently, the FRT-oriV retrofitted insertions were crossed with each other in pairs (KmRxCmR), using P1 phage transductions. The resulting CmRFRT-[28-65-kb]-KmRFRT strains were transformed with a plasmid expressing FLP and trfA genes from the tightly controlled Ptet promoter. Induction of this tightly repressed promoter by autoclaved chlortetracycline (cTc) resulted in the efficient excision and amplification of genomic fragments located between FRT sites, but only in productive strains, i.e. having two parallel FRTs. We have shown that genomic fragments of 28-, 40-, 50- and 65-kb were efficiently excised and amplified. Furthermore, we could convert non-productive strains (having FRTs in non-parallel orientation), to productive combination of parallel FRTs, because one of the FRT elements was flanked by two convergent loxP sites, and thus could be inverted by the Cre function delivered either by the P1 phage or by a specially constructed temperature-sensitive Plac-cre plasmid. Although several microbial genomes were recently sequenced, the described method will help in supplying large quantities of any genomic fragment (prepared without the conventional cloning and its artifacts) for refined sequence comparison among strains and species, and for further analysis of uncharacterized ORFs, various mutations, and regulatory elements or functions. The excised and circularized DNA fragments (plasmids) could be propagated like any other large plasmids but only in hosts that could supply the appropriate Rep function. Our original 'pop-out' method [Posfai et al. (1994) Nucleic Acids Res. 22, 2392-2398] was already employed for sequencing of the E. coli genome [Blattner et al. (1997) Science 277, 1453-1462]. Moreover, the Flp-mediated recombination between two FRT elements resulted in bacterial strains with large deletions (for parallel FRT orientations) or with large inversions (for inverted FRT orientations). PMID- 9858685 TI - Riboflavin transport by rabbit renal basolateral membrane vesicles. AB - The present study examined riboflavin (RF) uptake by isolated rabbit renal basolateral membrane (BLM). RF uptake was linear during the initial 10 seconds and leveled off thereafter with longer incubation. Studies on RF uptake as a function of incubation medium osmolarity indicated that the BLM RF uptake was the results of transport (approximately 45%) into the intravesicular space as well as binding (approximately 55%) to membrane surfaces. The RF binding to BLM was Na+ dependent so that replacement of Na+ by other cations eliminated the binding component of RF uptake. The process of BLM RF uptake was saturable as a function of substrate concentration and was significantly inhibited by cis-addition of its structural analogs, lumiflavin and lumichrome, indicating the involvement of a carrier-mediated process. The BLM RF uptake was affected by changes in extravesicular pH so that, as compared to pH 7.5, RF uptake was lower at pH 6.5 and higher at pH 8.5. The effect of extravesicular pH persisted when the transmembrane H+ gradient was dissipated by FCCP, indicating the direct effect of pH on BLM RF uptake. The BLM RF uptake was not affected by alterations of the transmembrane electrical potential, induced by either the presence of anions with different membrane permeability (Cl-=NO-3>SO-4>gluconate-) or using nigericin (10 microg/mg protein) with an outwardly or inwardly directed transmembrane K+ gradient. The BLM RF uptake was, however, inhibited by probenecid and p aminohippurate, and was enhanced by trans-RF. In summary, these results demonstrate the existence of a Na+-dependent BLM binding of RF and a membrane associated carrier system for RF uptake by renal BLM. PMID- 9858686 TI - Requirement for EphA receptor signaling in the segregation of Xenopus third and fourth arch neural crest cells. AB - We describe here the isolation of a full-length cDNA encoding a Xenopus orthologue of the mammalian EphA2 receptor tyrosine kinase and investigate its role in cranial neural crest migration. We show that the primary sites of Xenopus EphA2 expression are rhombomere 4 of the developing hindbrain, migratory cranial neural crest cells and mesoderm of the visceral arches. To interfere with EphA2 and related receptors during cranial neural crest migration, we took a dominant negative approach. Overexpression of kinase-deficient EphA2 receptor variants led to abnormal migration of cranial neural crest cells. Neural crest cells of the third arch were found to mismigrate posteriorly, resulting in the failure of third and fourth arch neural crest to separate into distinct streams. These defects could be rescued by expression of full-length EphA2 receptors. A comparison of the expression domains of EphA2-binding proteins mapped by receptor affinity probe (RAP) in situ staining with those for EphA2 receptors revealed co expression of ligands and receptors in the visceral arch mesenchyme. Taken together, these results suggest that EphA receptors may mediate attractive or adhesive signals during migration of cranial neural crest cells. PMID- 9858687 TI - Phospholipid acyl chain rotational dynamics are independent of headgroup structure in unilamellar vesicles containing binary mixtures of dioleoyl phosphatidylcholine and dioleoyl-phosphatidylethanolamine. AB - We have examined relationships between phospholipid headgroup structure and acyl chain dynamics, and their respective roles in modulating the physical properties of biological membranes. Fluorescence lifetime and anisotropy measurements were used to assess structural changes involving the lipid acyl chains in homogeneous populations of small and large unilamellar vesicles containing binary mixtures of dioleoyl-phosphatidylcholine (PC) and dioleoyl-phosphatidylethanolamine (PE) in the liquid-crystalline (Lalpha) phase. These measurements involve three different fluorescent lipid analogs containing diphenylhexatriene (DPH) linked to either a trimethylamine moiety (i.e., TMA-DPH) or the sn-1 position of monostearoyl phospholipids containing PC or PE headgroups (i.e., DPH-PC and DPH-PE). The average lifetimes, rotational correlation times, and order parameters associated with DPH-PC and DPH-PE are virtually identical, and are not affected by alterations in the PE content of the membrane. These results suggest that the average cross-sectional areas of the phospholipid acyl chains of DOPE and DOPC relative to the membrane normal are similar in these unilamellar vesicles. Since PC headgroups are larger than those of PE, differences in the relative orientation of the phosphocholine and phosphoethanolamine moieties relative to the membrane surface probably function to maintain optimal van der Waals contact interactions between acyl chains. On the other hand, the average lifetime associated with TMA-DPH, whose chromophoric group is near the membrane surface, increases with increasing PE content. The position of TMA-DPH relative to the membrane surface does not change, since the rotational dynamics of TMA-DPH are independent of the PE concentration. Therefore, alterations in the average lifetime of TMA-DPH results from polarity differences near the membrane surface at the level of the glycerol backbone. These results are discussed in terms of how differences in the average conformation of the glycerol backbones or phospholipid headgroups of PE and PC have the potential to regulate membrane function. PMID- 9858688 TI - A human cell system for detecting asbestos cytogenotoxicity in vitro. AB - Crocidolite, a carcinogenic asbestos in humans, specifically induces mesothelioma. We investigated the cytogenotoxic effects of crocidolite in a human mesothelioma cell line, MSTO211H, and a human promyelocytic leukemia cell line, HL60. Using confocal laser scanning microscopy, we found that the MSTO211H cells had phagocytotic activity, whereas the HL60 cells did not. In the MSTO211H cells, crocidolite decreased the cell population and increased the numbers of polynucleated cells (PN) and tetraploid cells, and increased the coefficients of variation (CV) of DNA contents in G0/G1 cells and the formation of 8 hydroxydeoxyguanosine. In contrast, crocidolite showed none of these cytogenotoxic effects in HL60 cells. To investigate the importance of phagocytosis in the cytogenotoxicity of crocidolite, we sorted the crocidolite phagocytosed cells from less-phagocytosed cells by fluorescence-activated cell sorting, and studied the differences in cytogenotoxicity between these two cell groups. We found significant increases in the numbers of PN and tetraploid cells and the CV in the crocidolite-phagocytosed cells compared to the less phagocytosed cells. These findings indicate that MSTO211H cells are susceptible to the cytogenotoxic effects of asbestos due to their phagocytotic activity, and that the MSTO211H cell line is suitable for the detection of such effects on human cells by asbestos and other materials which need to be phagocytosed to exert their toxicity. PMID- 9858689 TI - Cre/loxP-mediated in vivo excision of large segments from yeast genome and their amplification based on the 2microm plasmid-derived system. AB - In vivo excision and amplification of pre-determined, large genomic segments, directly from the genome of a natural host, provides an alternative to conventional cloning in foreign vectors. Using this approach, we have devised an in vivo procedure for excising large segments of Saccharomyces cerevisiae genome using Cre/loxP system of bacteriophage P1, followed by amplification of excised circles, as based on the yeast 2microm plasmid-derived ori and Flp/FRT machinery. To provide the excision and replication enzymes, trans-acting genes cre and FLP, which were under a very tight control of GAL1 and GAL10 promoters, respectively, were inserted by homologous recombination into the URA3 gene on chromosome V. Two parallel loxP sequences, which serve as the recognition sites for the Cre recombinase, were also integrated into the genome at pre-determined sites that are 50-100kb apart. Moreover, 2microm ori, REP3 and two inverted FRTs, which serve as a conditional replication system, were also integrated between the loxP sites. The strain carrying all these inserted elements was perfectly stable. Only after the induction by galactose of the Cre excision function, the genomic segment flanked by two loxP sites was excised and circularized. Applying this procedure, the 50-kb LEU2-YCR011c and 100-kb LEU2-YCR035c regions of chromosome III were successfully excised from the S. cerevisiae genome, whereas the 2microm ori, as aided by FRT/Flp, provided the amplification function. Such excised and amplified genomic segments can be used for the sequencing and functional analysis of any yeast genes. PMID- 9858690 TI - Triethylene glycol dimethacrylate induces large deletions in the hprt gene of V79 cells. AB - Acrylate esters are applied in industrial and consumer products often associated with polymers and resins. The difunctional methacrylate, triethylene glycol dimethacrylate (TEGDMA), is also frequently included in dental composite materials. Recently, mutagenicity testing of the compound revealed the induction of gene mutations at the hprt locus in V79 cell [H. Schweikl, G. Schmalz, K. Rackebrandt, The mutagenic activity of unpolymerized resin monomers in Salmonella typhimurium and V79 cells, Mutat. Res. 415 (1998) 119-130]. In the present study, TEGDMA caused a dose dependent increase of the number of micronuclei in V79 cells. Furthermore, the mutation spectra induced in exon sequences of the hprt gene in HPRT-deficient V79 cell clones were analyzed by the polymerase chain reaction (PCR). No DNA sequence deletions were observed in spontaneously occurring HPRT-deficient cell clones at the molecular level after PCR analysis, indicating that all spontaneous mutations were caused by point mutations. However, TEGDMA treated V79 cell cultures exhibited different mutation spectra. Only one cell clone among a total of 25 contained all exon sequences of the hprt gene. Large DNA sequences were deleted in 24 cell clones. Partial gene deletions occurred in four clones from exon 5 through 9, and exon 1 was not amplified in one cell clone. Exon sequences of the hprt gene were totally deleted in 19 HPRT deficient clones. The induction of mostly large deletions in the genome of mammalian cells, like the mutation spectra induced by TEGDMA in V79 cells here, is probably typical for crosslinking agents, including anticancer drugs. Identical types of mutations including chromosomal aberrations and the formation of micronuclei in vitro were observed for acrylates and methacrylates tested so far in various mutation assays. Therefore, we conclude by analogy that the induction of large DNA sequence deletions as shown here with the reactive dimethacrylate, triethylene glycol dimethacrylate, is probably common for acrylates and methacrylates. PMID- 9858691 TI - A new system to place single copies of genes, sites and lacZ fusions on the Escherichia coli chromosome. AB - To place a single-copy lacZ fusion on the E. coli chromosome, a method was developed based on in vivo homologous DNA recombination through P1 transduction. The fusions, initially constructed on plasmids, are crossed to lambdalacZ fusion vectors which are then lysogenized at the chromosomal lambda att site. The features of the new system are: (1) lambda lysogens carrying the fusion are made without regard for copy number; (2) P1 transduction from the lysogenic strain into an appropriate recipient generates the single-copy fusion; (3) The lacZ fusion has no prophage associated with it; (4) the lacZ fusion can be transferred by P1 transduction to other strains, simply by selecting for an antibiotic marker; (5) the system can be widely applied to construct single copies of any gene or site placed between bla and lacZ on the standard lacZ fusion plasmid vectors; and (6) the single-copy construct flanked by prophage att sites can be excised by site-specific recombination to generate non-replicating circular DNA of the clone or a cell cured of the construct. PMID- 9858692 TI - Assembly of the Kdp complex, the multi-subunit K+-transport ATPase of Escherichia coli. AB - Kdp, the high affinity ATP-driven K+-transport system of Escherichia coli, is a complex of the membrane-bound subunits KdpA, KdpB, KdpC and the small peptide KdpF. The assembly of this complex was studied by the analysis of mutants that expressed two of the three large subunits and inserted them into the cytoplasmic membrane. In the strains that do not express KdpC or KdpA the other two subunits did not copurify on dye-ligand affinity columns after solubilization with non ionic detergent. In the mutant lacking KdpB the other two subunits copurified under the same conditions. It is concluded that KdpC forms strong interactions with the KdpA subunit, serving to assemble and stabilise the Kdp complex. A structure in which KdpC could be one of the connecting links between the energy delivering subunit KdpB and the K+-transporting subunit KdpA is suggested by these data. PMID- 9858693 TI - Overlapping and non-overlapping Ptch2 expression with Shh during mouse embryogenesis. AB - In Drosophila, patched encodes a negative regulator of Hedgehog signaling. Biochemical experiments have demonstrated that vertebrate patched homologues might function as a Sonic hedgehog (Shh) receptor. In mice, two patched homologues, Ptch and Ptch2, have been identified. Sequence comparison have suggested that they might possess distinct properties in Shh signaling. In the developing tooth, hair and whisker, Shh and Ptch2 are co-expressed in the epithelium while Ptch is strongly expressed in the mesenchymal cells. We report here the chromosomal localization of Ptch2 and further analysis of Ptch2 expression. Throughout mouse development, the level of Ptch2 expression is significantly lower than that of Ptch. In early mouse embryos, Ptch and Ptch2 were found to be co-expressed in regions adjacent to Shh-expressing cells in the developing CNS. Similar to other epidermal structures, Shh and Ptch2 also show overlapping expression in the developing nasal gland and eyelids. Thus, during mouse development, Ptch2 is expressed in both Shh-producing and -nonproducing cells. PMID- 9858694 TI - Genomic DNA sequencing by SPEL-6 primer walking using hexamer ligation. AB - DNA sequencing by SPEL-6 (Sequential Primer Elongation by Ligation of 6-mers) primer walking is based on the rapid assembly of true primers by ligation of several (three to 10) contiguous hexamers complementary to a DNA template saturated with Escherichia coli single-stranded DNA-binding protein. To prove the usefulness and to check the reliability of this method, a 3-kb DNA fragment carrying the genes encoding the EcoVIII restriction-modification (RM) system was sequenced with low redundancy (2.8). The use of both single-stranded (ss) and double-stranded (ds) DNA templates was compared. For this project, 27 primers were assembled by hexamer ligation to form 18-30-nt strings of three to five hexamers. Each primer was designed based on nucleotide sequence determined in a previous run, and was produced in a matter of minutes. The overall length of the easily readable sequencing ladders was about 300-450nt. We found that strong secondary structures in the ss DNA tend to interfere with its template function for the primer assembly by hexamer ligation, especially when they overlap the 3' end of such a primer. This was easily overcome either by avoiding such hairpin regions or by using longer strings of hexamers, since we show that their ligation is highly cooperative, and ligation efficiency increases with the length of the string (). Some general rules for successful primer assembly and prospects for using the SPEL-6 method for large-scale, fully automated fluorescent sequencing of large genomes are discussed. PMID- 9858695 TI - Isolation, expression and regulation of a zebrafish paraxis homologue. AB - The formation of somites involves the subdivision of segmented presomitic mesoderm into segmentally arranged somite blocks. In mice and chicks, the basic helix-loop-helix (bHLH) gene, paraxis, is involved in this process. Here, we report the isolation of a zebrafish homologue of paraxis, par1. par1 is expressed in presomitic paraxial mesoderm from late gastrula stages, and expression is maintained in ventrolateral cells after somite formation. In spt- embryos, par1 expression is both delayed and severely reduced whereas in flh- embryos, ectopic transcripts are detected in axial mesoderm. Spatial regulation of par1 expression within the somites is affected in several mutants with defects in axial midline tissues. PMID- 9858696 TI - Human proteolipid protein (PLP) mediates winding and adhesion of phospholipid membranes but prevents their fusion. AB - Proteolipid protein (PLP or lipophilin) is a highly conserved, strongly hydrophobic, integral membrane protein, and is the major protein component of central nervous system myelin. Although PLP has been implicated in many functions, its in vivo role is still uncertain. Here, we report the investigation of PLP's putative adhesive function using purified PLP and reconstituted phospholipid vesicles made of either 100% phosphatidylcholine (PC), or a mixture of 92% PC and 8% phosphatidylserine (PS), by weight. PLP-induced changes in the phospholipid bilayer surfaces were directly examined by transmission electron microscopy. We found that upon the introduction of PLP, larger lipid vesicles became smaller and unilamellar. At the PLP:lipid molar ratio of 1:20, vesicle membranes rolled onto themselves forming 'croissant'-like structures that subsequently adhered to each other. The phenomena of PLP-induced bilayer rolling and adhesion were dependent on the concentration of PLP and the period of incubation, but were independent of the presence of calcium and types of phospholipids (PC or PC:PS). Furthermore, the presence of PLP in the lipid bilayers prevented the fusion of membranes. These findings show that PLP can induce membrane 'winding' while preventing the fusion of adjacent lipid bilayers. Hence, our data provide direct evidence for PLP's suspected function of membrane adhesion, and also suggest that PLP could potentially play a role in the formation of the myelin sheath. PMID- 9858697 TI - mCelsr1 is an evolutionarily conserved seven-pass transmembrane receptor and is expressed during mouse embryonic development. AB - Mcelsr1 encodes a protein of 3034 amino acids predicted to contain seven membrane spanning domains having homology to a group of peptide hormone binding G-protein coupled receptors. Its extracellular domain comprises epidermal growth factor like repeats, laminin A G-domains and cadherin repeats. Homologous genes have been identified in C. elegans and D. melanogaster suggesting that the Celsr gene family is ancient. mCelsr1 mRNA expression precedes gastrulation, is subsequently restricted primarily to ectodermal derivatives and is tightly regulated in the developing central nervous system (CNS). We observe segmentally-restricted gene expression in the developing hindbrain and in the spinal cord dynamic dorso ventrally restricted 'stripes' of expression. PMID- 9858698 TI - Recombination in phage lambda: one geneticist's historical perspective. AB - Several features of bacteriophage lambda suit it for the study of genetic recombination. Central among them are those that make it possible to correlate inheritance of DNA with the inheritance of information encoded by DNA through density-label equilibrium centrifugation. Such studies have revealed relationships between DNA replication and recombination, have identified roles for double-strand breaks in the initiation of recombination, and have elucidated the role of the recombination-stimulating sequence, chi. PMID- 9858700 TI - Interactions of a vesicular stomatitis virus G protein fragment with phosphatidylserine: NMR and fluorescence studies. AB - The interaction of a 19 amino acid vesicular stomatitis virus G protein fragment (GTWLNPGFPPQSCGYATVT) with phosphatidylserine-containing model membranes was investigated using solution-phase 1d and 2d 1H NMR spectroscopy and intrinsic tryptophan fluorescence. Results of these studies show that this peptide interacts with model membranes containing negatively charged phospholipids. The interaction is modulated by both ionic and hydrophobic factors and appears to be dependent on the fluidity and lipid packing of the target bilayer. The data further suggest the existence of two isomeric forms of this peptide, which react differentially with model membranes. Upon binding, 2d 1H NOESY and tryptophan fluorescence data indicate penetration of the tryptophan residue into the bilayer. A model is proposed for the interaction of the peptide with model membranes, consistent with the experimental findings. PMID- 9858701 TI - From bacteriophage lambda to nonlinear dynamics. AB - Thirty years ago, Waclaw and myself were working on the same organism and had the same type of interest. Since that happy time, our trajectories have diverged so much that I am afraid any paper I might offer for his jubilee would seem completely strange and irrelevant to him and as well to his colleagues. For this reason, I decided rather to offer Waclaw, as a modest present, a product of my recent work which I consider, perhaps presumptuously, as an 'oeuvre d'art' (of course, this qualification is not supposed to apply to this paper as a whole, just to the figure). Please note above the expressions 'divergent trajectories' and 'strange'; we will meet them again below in a different context. PMID- 9858699 TI - Direct binding between two PDZ domain proteins Canoe and ZO-1 and their roles in regulation of the jun N-terminal kinase pathway in Drosophila morphogenesis. AB - During Drosophila embryogenesis, the ventral epidermis dorsally expands and the left and right epithelial sheets meet and fuse along the dorsal midline. For this dorsal closure to occur, two PDZ domain proteins, Cno and ZO-1, are required. The dorsal epidermis remains open when the expression of ZO-1 and Cno are reduced simultaneously by hypomorphic mutations in the relevant loci. ZO-1 and Cno colocalize at adherens junctions in embryonic epithelia, and form a protein complex upon binding to each other. Genetic analysis showed that Cno is involved in the Jun N-terminal kinase (JNK) pathway for dorsal closure, as a modulator acting upstream of, or in parallel with, the small GTPase Drac1. The ZO-1-Cno complex may be involved in dynamic changes in cytoskeletal organization and cell adhesion during morphogenetic events associated with dorsal closure in the Drosophila embryo. PMID- 9858702 TI - Arrangement and functional identification of genes in the regulatory region of lambdoid phage H-19B, a carrier of a Shiga-like toxin. AB - H-19B is a lambdoid phage that carries the genes (stx-I) encoding the two toxin subunits of a Shiga-like toxin; Escherichia coli lysogens of H-19B are converted to toxin producers. Based on the determination of a 17-kb region of the H-19B genome and functional studies, we have identified the early regulatory region and associated genes of H-19B, as well as the location of the late regulatory region and the toxin and lysis genes. A comparative analysis of the sequence of the H 19B genome reveals the presence of ORFs and genes found in analogous positions on the genomes of a number of other lambdoid phages. A cloned genomic fragment that confers immunity to an infecting H-19B phage contains an ORF of an analogous size and genomic location for a repressor gene, adjacent to a putative operator region. The lambda replication genes, O and P, are conserved in H-19B except for a 39-bp insert in the O gene creating two new O protein-binding sites in the origin of replication (ori), giving H-19B six binding sites as opposed to the four sites found in lambda. We identify ORFs and sequences involved in transcriptional regulation encoding N-like antitermination systems like those found in other lambdoid phages and nearly identical to sequences found in phage HK97. Our functional studies show that these sequences support antitermination even though they contain significant differences from those of other lambdoid phages. We also identify ORFs and sequences analogous to the Q-p'R late antiterminators-promoters found in other lambdoid phages. The Shiga-like stx-I genes are located directly downstream of the promoter, p'R, for the late genes, and upstream of the lysis genes. PMID- 9858704 TI - The turgor sensor KdpD of Escherichia coli is a homodimer. AB - Escherichia coli responds to K+-limitation or high osmolarity by induction of the kdpFABC operon coding for the high affinity K+-translocating KdpFABC complex. Expression of the corresponding operon is controlled by the membrane-bound sensor kinase KdpD and the cytoplasmic response regulator KdpE. Here, we examine the oligomeric state of KdpD. KdpD-His673-->Gln and KdpD-Asn788-->Asp are kinase inactive. When the corresponding genes are coexpressed, the resulting KdpD protein regains kinase activity in vitro, suggesting that the functional state of KdpD is at least a dimer and that the kinase reaction is a result of a trans phosphorylation between two monomers. Furthermore, coexpression of kdpD-6His and kdpD-(Delta128-391) leads to stable heterooligomers that can bind to Ni-NTA agarose and that are coeluted. Purified and solubilized KdpD-6His has been electrophoresed in blue native polyacrylamide gels (BN-PAGE), and unphosphorylated and phosphorylated KdpD resulted in the same band pattern suggesting that the oligomeric state of KdpD does not change upon phosphorylation. In addition, determination of the molecular masses of KdpD-6His and KdpD-6His approximately 32P by gel filtration reveals a value of 245 kDa for both forms of the protein. The Stokes radius is determined to be 5.4 nm. Sucrose gradient sedimentation analysis of KdpD-6His results in a molecular mass of 289 kDa. The calculated molecular mass of a KdpD-6His monomer is 99.6 kDa. Considering the detergent bound to KdpD the obtained data reveal that KdpD is a homodimer and there is no change in the oligomeric state upon activation. Crosslinking experiments with single Cys KdpD molecules indicate that there is a close contact between the monomers in the transmitter as well as in transmembrane domain 1. BN-PAGE of solubilized and purified KdpD-6His devoid of Cys residues demonstrates that Cys residues do not contribute to the stabilization of the dimer. PMID- 9858703 TI - Gal4 in the Drosophila female germline. AB - The modular Gal4 system has proven to be an extremely useful tool for conditional gene expression in Drosophila. One limitation has been the inability of the system to work in the female germline. A modified Gal4 system that works throughout oogenesis is presented here. To achieve germline expression, it was critical to change the basal promoter and 3'-UTR in the Gal4-responsive expression vector (generating UASp). Basal promoters and heterologous 3'-UTRs are often considered neutral, but as shown here, can endow qualitative tissue specificity to a chimeric transcript. The modified Gal4 system was used to investigate the role of the Drosophila FGF homologue branchless, ligand for the FGF receptor breathless, in border cell migration. FGF signaling guides tracheal cell migration in the embryo. However, misexpression of branchless in the ovary had no effect on border cell migration. Thus border cells and tracheal cells appear to be guided differently. PMID- 9858705 TI - Acquired mutations in phage lambda genes O or P that enable constitutive expression of a cryptic lambdaN+cI[Ts]cro- prophage in E. coli cells shifted from 30 degreesC to 42 degreesC, accompanied by loss of immlambda and Rex+ phenotypes and emergence of a non-immune exclusion-state. AB - The majority of bacteria, which carry the N+-cI857[Ts]-cro--O+-P+ fragment of lambda genome, are killed when derepressed by shifting from 30 degreesC to 42 degreesC. Among rare survivors, we observed a proportion of colony-forming units (cfu) that retained the typical immlambda-immunity phenotype when grown at 30 degreesC; however, when shifted from 30 degreesC to 42 degreesC, they lost lambda immunity and acquired a non-immune exclusion-state (Nie phenotype). We also found that the immlambda survivor cfu quickly lost their Rex+ exclusion phenotype (as measured by T4rII plating inhibition) when shifted from 30 degreesC to 42 degreesC, even though they produced CII, which stimulates pE-cI-rexA-rexB transcription. The Nie phenotype was characterized by an inhibition of plating of the homoimmune phage, lambdawt, and the heteroimmune phage, lambdaimm434. However, lambdavir and spontaneous mutants of lambdawt (lambdase mutations localized within oR) escaped the Nie exclusion-state and plated efficiently on lawns of Nie cfu at 42 degreesC. Thus, we examined the scope of the Nie exclusion state toward lambda mutants blocked for lysogeny, and lambda hybrids substituted for immunity or replication genes. Phage like lambdawt, competent for lysogeny, were severely excluded compared to some mutants of lambda defective for lysogeny. Among this latter type, there was high variance in the Nie exclusion of various cI mutants; some of which were not excluded. The Nie exclusion-state was attributed to the constitutive expression of the defective lambda fragment in the survivor cfu, made possible by the acquired replication defect(s). We characterized, both genetically and physically, the mutations in the defective integrated lambda prophage that permitted growth of the survivor cfu at 42 degreesC. In five of seven survivor cfu, we identified IS2 insertions within lambda genes O and P that can block replication initiation from the lambda fragment. The remaining survivor cfu had multiple base substitutions within the C terminal end of O and N-terminal half of P, the majority of which were silent. In some of these mutants, either an ochre nonsense mutation or a single-base frameshift deletion inactivated P. PMID- 9858706 TI - Fgf-R3 is expressed in a subset of chicken spinal motorneurons. AB - The expression pattern of the fibroblast growth factor receptor Fgf-R1, R2 and R3 genes was studied in the chicken spinal cord using in situ hybridization (ISH). Unlike Fgf-R1 which is widely expressed in motoneurons, Fgf-R3 is expressed in a subset of motoneurons in the medial subdivision of the median motor column (MMCm) that also express Islet-1 and Lim-3. The motoneuron identity of the labelled cells was confirmed by double ISH and by single cell RT-PCR. Interestingly, E3.5 spinal cord motoneurons do not express Fgf-R3, suggesting the expression of Fgf R3 in motoneurons begins with axonal growth. PMID- 9858707 TI - Transcriptional control of Drosophila bicoid by Serendipity delta: cooperative binding sites, promoter context, and co-evolution. AB - Concentration of maternal BICOID (BCD) establishes the anterior pattern in the Drosophila embryo. Successive deletions in the bcd promoter allowed us to localize an enhancer sequence in the 5'-UTR and a down-regulating element downstream of the ATG initiator codon, and identify a 49 bp region sufficient to drive transcription of a reporter gene specifically in nurse cells. This fragment contains two binding sites for the Serendipity (Sry) d zinc finger activator, that mediate its cooperative binding. Both sites (sdbs) are essential for bcd expression. Further analysis showed that the bcd promoter configuration is decisive for Sry d activating function. Replacement of sdbs by binding sites for Sry b, the Sry d paralog, restores bcd transcription in sry d mutant ovaries, demonstrating that the functional divergence between these two proteins during evolution was mainly driven by changes in their DNA-specific recognition properties, resulting in the control of separate developmental pathways. PMID- 9858708 TI - Rotational dynamics of spin-labelled surfactant-associated proteins SP-B and SP-C in dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylglycerol bilayers. AB - Pulmonary surfactant proteins SP-B and SP-C have been isolated from porcine lungs and selectively labelled with 2,2,6, 6-tetramethylpiperidine-N-oxyl (TEMPO) isothiocyanate at their N-terminal amine ends, to analyse the mobility of both proteins on the nanosecond time scale using electron spin resonance (ESR) spectroscopy. Reconstitution of the labelled forms of these proteins in bilayers of dipalmitoylphosphatidylcholine (DPPC) or dipalmitoylphosphatidylglycerol (DPPG) results in much broader and anisotropic ESR spectra, indicating a large restriction in rotational mobility of the protein-attached probe when inserted in membranes. Distinctive differences were found between the ESR spectra of the two polypeptides, that were consistent with intrinsic differences in mode of interaction of SP-B and SP-C with phospholipid bilayers. The mobility of the protein spin probes was sensitive to temperature on the time scale of conventional spin-label ESR. Both proteins, TEMPO-SP-B and TEMPO-SP-C, showed considerable increases in mobility at temperatures above the pretransition of pure DPPC. Finally, the mobility of the spin probes attached to both SP-B and SP C was more restricted in DPPG than in DPPC bilayers, demonstrating that electrostatic interactions of the positively charged residues at the protein surface influence the rotational dynamics of the proteins in anionic lipid bilayers. Although some residual segmental mobility of the thiourea-linked probes cannot be discounted, the results clearly reflect preferential differences in overall protein dynamics in gel and fluid phases of the two phospholipids that could be important for the biophysical properties of surfactant bilayers and monolayers. PMID- 9858709 TI - Mutational analysis of a satellite phage activator. AB - The late gene activator, Delta, of satellite phage P4 is more efficient than the Delta of satellite phage phiR73 in utilizing a P2 helper prophage that lacks an activator (ogr) gene. Analysis of P4 Delta is complicated by the fact that this protein contains two tandem phiR73 Delta-like domains. We performed a mutational analysis of phiR73 Delta, in order to select mutations that might not be found using P4 Delta. The host RNA polymerase alpha subunit mutation rpoA155 (L289F) blocks the growth of P2, P4, and P4 carrying the delta gene of phiR73. A mutant of this latter phage that can grow in the presence of rpoA155 carries a V19M mutation in phiR73 Delta. This suggests that aa 19 contacts RNA polymerase, in addition to the aa residues 13, 42 and 44, that have been implicated in interactions with RNA polymerase by previous mutational analyses of P2 ogr and P4 delta. In corroboration of the proposed role of the regions at aa residues 19, 42, and 44, we found phiR73 Delta mutations in these regions that showed a reduced activation of late gene expression, but a normal ability to bind to late gene promoters. All activators in the Delta class contain four Cys residues that bind Zn2+. Mutation of these aa residues in phiR73 Delta eliminated late gene activation. Spectroscopic analysis of these mutant proteins revealed that they were unable to bind Zn2+. Histidine residues were substituted for two of the Cys residues (C30 and C35), changing a C2C2 type Zn-binding motif to a C2H2 motif. Although His residues are used in coordinating Zn2+ in other proteins, these His substitutions resulted in complete loss of activity and the inability to bind Zn2+. PMID- 9858710 TI - Bacteriophage phi29 early protein p17 is conditionally required for the first rounds of viral DNA replication. AB - The gene 17 of the Bacillus subtilis phage phi29 is known to be involved in the viral DNA replication in vivo. In this paper, we show that the presence of protein p17 is required when phage infection occurs at a low multiplicity of infection (moi), which is probably the natural condition for infection, but is dispensable at a high moi. Gene 17 has been cloned in an Escherichia coli expression vector and protein p17 purified. A stimulatory effect of protein p17 was demonstrated under in vitro conditions required to amplify phi29 DNA, starting with a low amount of input DNA. We propose that p17, which is synthesized early after infection, is required at the very beginning of the phage amplification, conditions in which a low number of viral DNA molecules enter the host cell, possibly to recruit the limiting amount of initiation factors at the replication origins. Once the infection process is established and the other replication proteins reach optimal concentration, p17 becomes dispensable. PMID- 9858712 TI - Interaction of the neuronal marker dye FM1-43 with lipid membranes. Thermodynamics and lipid ordering. AB - The fluorescent dye FM1-43 labels nerve terminals in an activity-dependent fashion and has been found increasingly useful in exploring the exo- and endocytosis of synaptic vesicles and other cells by fluorescence methods. The dye distributes between the aqueous phase and the lipid membrane but the physical chemical parameters characterizing the adsorption/partition equilibrium have not yet been determined. Fluorescence spectroscopy alone is not sufficient for a detailed elucidation of the adsorption mechanism since the method can be applied only in a rather narrow low-concentration window. In addition to fluorescence spectroscopy, we have therefore employed high sensitivity isothermal titration calorimetry (ITC) and deuterium magnetic resonance (2H-NMR). ITC allows the measurement of the adsorption isotherm up to 100 microM dye concentration whereas 2H-NMR provides information on the location of the dye with respect to the plane of the membrane. Dye adsorption/partition isotherms were measured for neutral and negatively-charged phospholipid vesicles. A non-linear dependence between the extent of adsorption and the free dye concentration was observed. Though the adsorption was mainly driven by the insertion of the non-polar part of the dye into the hydrophobic membrane interior, the adsorption equilibrium was further modulated by an electrostatic attraction/repulsion interaction of the cationic dye (z=+2) with the membrane surface. The Gouy-Chapman theory was employed to separate electrostatic and hydrophobic effects. After correcting for electrostatic effects, the dye-membrane interaction could be described by a simple partition equilibrium (Xb=Kcdye) with a partition constant of 103-104 M-1, a partition enthalpy of DeltaH=-2.0 kcal/mol and a free energy of binding of DeltaG=-7.8 kcal/mol. The insertion of FM1-43 into lipid membranes at room temperature is thus an entropy-driven reaction following the classical hydrophobic effect. Deuterium nuclear magnetic resonance provided insight into the structural changes of the lipid bilayer induced by the insertion of FM1-43. The dye disturbed the packing of the fatty acyl chains and decreased the fatty acyl chain order. FM1-43 also induced a conformational change in the phosphocholine headgroup. The -P-N+ dipole was parallel to the membrane surface in the absence of dye and was rotated with its positive end towards the water phase upon dye insertion. The extent of rotation was, however, much smaller than that induced by other cationic molecules of similar charge, suggesting an alignment of FM1-43 such that the POPC phosphate group is sandwiched by the two quaternary FM1-43 ammonium groups. In such an arrangement the two cationic charges counteract each other in a rotation of the -P-N+ dipole. PMID- 9858711 TI - A novel growth differentiation factor-9 (GDF-9) related factor is co-expressed with GDF-9 in mouse oocytes during folliculogenesis. AB - Growth differentiation factor-9 (GDF-9) is a transforming growth factor-b (TGF-b) family member which is expressed in the oocytes in mouse ovaries (McGrath, S.A., Esquela, A.F., Lee, S.J., 1995. Oocyte-specific expression of growth/differentiation factor-9. Mol. Endocrinol. 9, 131-136). GDF-9 is indispensable for normal folliculogenesis since female mice deficient for the GDF 9 gene are infertile due to an arrest of follicular growth at the primary follicle stage (Dong, J., Albertini, D.F., Nishimori, K., Kumar, T.R. , Lu, N., Matzuk, M.M., 1996. Growth differentiation factor-9 is required during early ovarian folliculogenesis. Nature 383, 531-535). We searched the GenBank Expressed Sequence Tag (EST) database with the mouse GDF-9 cDNA sequence, and identified from a mouse 2-cell embryo library an EST cDNA that encodes a putative member of the TGF-b superfamily, and named it as GDF-9B. Northern blot hybridization analyses of mouse ovaries revealed a single transcript of approximately 4.0 kilobases (kb) for GDF-9B and of 2.0 kb for GDF-9. We cloned by reverse transcription-polymerase chain reaction from mouse ovarian RNA a partial 821-base pair GDF-9B cDNA that spans the sequence encoding the putative mature region of GDF-9B. The COOH-terminal region of GDF-9B appears to be 53% homologous to GDF-9. Moreover, like GDF-9, GDF-9B lacks the cysteine residue needed for the covalent dimerization of several TGF-b family members. Using in situ hybridization analysis, we demonstrate that GDF-9B and GDF-9 mRNAs are co-localized in the oocyte. We also show that GDF-9B and GDF-9 genes are co-ordinately expressed during follicular development. PMID- 9858713 TI - Drosophila ciD encodes a hybrid Pangolin/Cubitus interruptus protein that diverts the Wingless into the Hedgehog signaling pathway. AB - The Hedgehog (Hh) and Wingless (Wg) signaling pathways play important roles in animal development. The activities of the two pathways depend on each other during Drosophila embryogenesis. In the embryonic segment, Wg is required in anterior cells to sustain Hh secretion in adjacent posterior cells. Hh input in turn is necessary for anterior cells to maintain wg expression. The Hh and Wg pathways are mediated by the transcription factors Cubitus interruptus (Ci) and Pangolin/TCF (Pan), respectively. Coincidentally, pan and ci are adjacent genes on the fourth chromosome in a head-to-head orientation. Our genetic and in situ hybridization data indicate that ciD is a mutation affecting both ci and pan. Molecular analysis revealed that the ciD allele is caused by an inversion event that swapped the promoter regions and the first exons of the two genes. The ci gene in ciD is controlled by the ubiquitous pan promoter and encodes a hybrid Ci protein that carries the N-terminal region of Pan. This domain has previously been shown to bind to the b-catenin homolog Armadillo (Arm), raising the possibility that Wg input, in addition to Hh input, modulates the activity of the hybrid CiD protein. Indeed, we found that Wg signaling induces the expression of the Hh target gene patched (ptc) in ciD animals. We provide evidence that this effect depends on the ability of the CiD protein to bind Arm. Genetic and molecular data indicate that wild-type Pan and CiD compete for binding to Arm, leading to a compromised transduction of the Wg signal in heterozygous ciD/+ animals and to a dramatic enhancement of the gain-of-function activity of CiD in homozygous mutants. Thus, the Hh and the Wg pathways are affected by the ciD mutation, and the CiD fusion protein integrates the activities of both. PMID- 9858715 TI - Mechanical aspects of membrane thermodynamics. Estimation of the mechanical properties of lipid membranes close to the chain melting transition from calorimetry. AB - Changes in the internal energy of lipids with temperature are related to both lipid volume and area changes. Close to the chain melting transition of lipid bilayers volume and enthalpy fluctuations generally follow proportional functions. This makes it possible to calculate the relationship between membrane excess heat capacity with lipid volume, area compressibility and the membrane bending modulus, if the area fluctuations of the two monolayers are assumed to be mainly decoupled. Thus, compressibility and elasticity display pronounced maxima at the chain melting transition. These maxima can also be related to pronounced minima of the sound velocity in the lipid transition range, which were found in ultrasonic experiments. In the present study heat capacity profiles and volume changes were obtained. The compressibilities and the bending modulus were then deduced from the specific heat. The relevance of these findings for structural transitions and for the curvature dependence of heat capacities is discussed. PMID- 9858714 TI - Several new bacteriophage T4 genes, mapped by sequencing deletion endpoints between genes 56 (dCTPase) and dda (a DNA-dependent ATPase-helicase) modulate transcription. AB - We have analyzed DNA of wild-type T4 and of 13 independent large viable deletions isolated by Homyk and Weil (Virology 61 (1974) 505-523) and by Little (Virology 53 (1973) 47-59), by sequencing, cloning, and expression studies. The deletions can be explained by illegitimate recombination between short (4- to 15-bp) ectopic repeats. In four deletions, adjacent regions are partially homologous, and in at least one of them, the base adjacent to the overlap was deleted during recombination. The sequence 5'-GGGC, which has not been associated with T4 deletions in other map regions, occurs within three repeats, and near the repeats in four more of the 13 deletions. Five previously named genes, 69, soc, mrh, modA, and dda were mapped relative to the deletion endpoints. Nine additional ORFs were found interspersed between them. One of these shares some similarities with mrh (modulates rpoH; Frazier and Mosig, Gene 88 (1990) 7-14), and another one resembles modA (coding for an ADP-ribosyl-transferase that modifies RNA polymerase alpha subunits, Skorko et al., Eur. J. Biochem. 79 (1977) 55-66) respectively. We found that the host's heat shock sigma factor, sigma32, is phosphorylated, and that Mrh protein modulates this phosphorylation. The ORF dda.9 downstream of mrh has a patchy similarity with conserved C-terminal segments (motifs) of sigma32; therefore, we call it srh. Another ORF, dda.2 located between modA and dda, shares sequence similarity with sigma70, and we call it srd. We consider the possibility that Srh and Srd act as decoys for sigma32, or sigma70, respectively. Expression of several of the ORFs from cloned DNA appears to be toxic to the host bacteria. Mutant clones only could be constructed from gene 69 and from modA. Moreover, dda.2 (srd)-containing bacteria grow extremely slowly, and they form filaments in liquid cultures. Clones carrying mrh and srh show less severe filamentation. Our results highlight the importance of 'non-essential' genes for phage development and evolution. PMID- 9858716 TI - Maternal Nanos regulates zygotic gene expression in germline progenitors of Drosophila melanogaster. AB - Maternal Nanos (Nos) protein is required for germline development in Drosophila embryos. Here we show that Nos regulates zygotic gene expression in the germline progenitors, or pole cells. In order to probe the gene expression in pole cells, we screened ten enhancer-trap lines which showed beta-gal expression in pole cells. All of these enhancer-trap markers were fully activated in pole cells after their migration to the embryonic gonads. In the pole cells lacking Nos, the expression of nine out of ten enhancer-trap markers was affected. Among nine markers, five (Type-A) were prematurely expressed in the pole cells during the course of their migration. The expression of other four markers (Type-B) initiated correctly after pole-cell migration, but their expression was significantly reduced. Thus, we conclude that the maternal Nos plays a dual role in zygotic gene regulation in pole cells: to define the stages of expression for Type-A markers, and to enhance expression for Type-B markers. Contrary to our results, "Heller and Steinmann-Zwicky (1998)" have recently reported that no premature expression of Type-A markers occurs in the pole cells of embryos derived from nos mutant females. This discrepancy is due to the difference in the nos mutant alleles used for these analyses. We used the much stronger allele, nosBN. PMID- 9858717 TI - Characterization of the nuclear localization signal in the avian sarcoma virus integrase. AB - A sequence of 21 amino acids (aa) in the C-terminal region of the 286-aa avian sarcoma virus (ASV) integrase (IN) protein has been shown previously to mediate nuclear localization of both IN and beta-galactosidase (betaGal) protein fused to it. This karyophilic sequence includes a high proportion of prolines and residues with basic side chains. In this report, site-directed mutagenesis was used to introduce single aa substitutions of several of these residues. Indirect immunofluorescence showed that IN-betaGal fusion constructs with Ala substitutions for sequence constituents K206, P215, K225 or R227 had lost the exclusive nuclear localization capability of the wild-type fusion. A fusion protein with the conservative substitution K206R retained the nuclear localization capacity. The site-specific substitutions that reduced karyophilic activity had no effect on the processing or joining activities of IN in vitro. However, the introduction of three of the four Ala codon substitutions into viral DNA clones caused a significant delay in viral replication following transfection of cycling chicken embryo fibroblasts. These results are consistent with a possible role for ASV IN in nuclear targeting. PMID- 9858719 TI - Amphiphilic and hydrophilic forms of acetylcholinesterase from sheep platelets. AB - Acetylcholinesterase (AChE, EC 3.1.1.7) was extracted from sheep platelets by successive homogenizations, yielding low-salt soluble (LSS), high-salt soluble (HSS) and detergent-soluble (DS) fractions. These accounted, respectively, for about 30%, 7% and 60% of total AChE activity. Applications of hydrophobic chromatography on phenyl-agarose to three solubilized fractions revealed that hydrophilic forms were almost exclusively located in the LSS fraction ( approximately 27% of total AChE), whereas most amphiphilic forms were present in DS extracts ( approximately 59% of total AChE), the remaining forms being distributed among aqueous soluble fractions. Enzyme molecular forms in the solubilized extracts were identified by centrifugation in 5-20% sucrose gradients containing Triton X-100 or Brij 97 to differentiate between hydrophilic or amphiphilic species. A predominance of hydrophilic dimeric forms ( approximately 22%), with small amounts of hydrophilic monomers (5%) and amphiphilic dimers and monomers (3%), was found in soluble AChE (LSS fraction). Amphiphilic AChE forms extracted in the HSS and DS fractions had a single peak in the sedimentation profiles with sedimentation coefficients of about 6S in gradients with Triton X 100; these were slightly shifted in the presence of Brij 97. After treatment with dithiothreitol, this molecular form solubilized in DS was converted to another molecular form with a lower sedimentation coefficient. Our results show that amphiphilic globular dimers are the dominant molecular form in sheep platelet AChE, suggesting a partial conversion of this membrane-bound form into soluble dimers and monomers, mainly with a hydrophilic character, through the action of either endogenous proteases and phospholipases or residual endogenous reducing agents. PMID- 9858718 TI - Distinct expression patterns of notch family receptors and ligands during development of the mammalian inner ear. AB - The cochlea and vestibular structures of the inner ear labyrinth develop from the otic capsule via step-wise regional and cell fate specification. Each inner ear structure contains a sensory epithelium, composed of hair cells, the mechanosensory transducers, and supporting cells. We examined the spatio-temporal expression of genes in the Notch signaling pathway, Notch receptors (Notch1-4) and two ligands, Jagged1 and Delta1, in the developing mammalian inner ear. Our results show that Notch1 and Jagged1 are first expressed in the otic vesicle, likely involved in differentiation of the VIIIth nerve ganglion neurons, and subsequently within the inner ear sensory epithelia, temporally coincident with initial hair cell differentiation. Notch1 expression is specific to hair cells and Jagged1 to supporting cells. Their expression persists into adult. Notch2, Notch3, Notch4, and Delta1 are excluded from the inner ear epithelia. These data support the hypothesis that Notch signaling is involved in hair cell differentiation during inner ear morphogenesis. PMID- 9858721 TI - Chicken anemia virus strains with a mutated enhancer/promoter region share reduced virus spread and cytopathogenicity. AB - Plasmid pCAV/E contains an infectious cloned double-stranded CAV (chicken anemia virus) DNA genome (Noteborn et al., J. Virol. 65 (1991) 3131-3139). We have constructed mutated CAV genomes by introducing mutations into the CAV promoter/enhancer region of pCAV/E. Various mutated CAV strains were functional and had a smaller cytopathogenic effect in chicken T cells than wild-type CAV. In particular, mutations within the '12-bp insert' of the promoter/enhancer region had this effect. PCR and sequence analysis showed that the CAV mutants were stable under cell-culture conditions. Southern-blot analysis showed that all replication DNA intermediates were normally formed by the CAV mutants. All viable mutant CAV strains were able to produce a neutralizing conformational epitope, which implies that they can trigger the required protective immune response. These features make these mutant CAV strains potential candidates for the development of an attenuated CAV vaccine. PMID- 9858720 TI - Embryonic expression and activity of doughnut, a second RYK homolog in Drosophila. AB - In the Drosophila embryo, a subset of muscles require expression and function of the RYK subfamily RTK gene derailed (drl) for correct attachment. We have isolated a second RYK homolog, doughnut (dnt), from Drosophila. The DNT protein exhibits 60% amino acid identity to DRL, and is structurally as similar to the mammalian RYK proteins as is DRL, indicating an ancient duplication event. dnt is expressed in dynamic patterns in the embryonic epidermis, being found at high level in epithelia adjacent to cells that are invaginating into the interior of the embryo, including ventral furrow, cephalic furrow, fore- and hindgut, optic lobe and tracheal pits. dnt is capable of a partial rescue of the muscle attachment defect of drl-/- embryos, indicating that it encodes a receptor with a related and significantly overlapping biochemical function. PMID- 9858722 TI - Pax3 and Pax7 are expressed in commissural neurons and restrict ventral neuronal identity in the spinal cord. AB - Pax3 and Pax7 are transcription factors sharing high sequence identity and overlapping patterns of expression in particular in the dorsal spinal cord. Analysis of Pax3 and Pax7 double mutant mice demonstrates that both genes share redundant functions to restrict ventral neuronal identity in the spinal cord. In their absence, the En1 expression domain is expanded dorsally but that of Evx1 is not affected. In addition, Pax3 and Pax7 are expressed in commissural neurons and double mutant embryos exhibit highly reduced ventral commissure. Our findings reveal two distinct regulatory pathways for spinal cord neurogenesis, only one of which is dependent on Pax3/7 and 6. PMID- 9858723 TI - The lipid/protein interface as a target site for general anesthetics: a multiple site kinetic analysis of synaptosomal Ca2+-ATPase. AB - There is a long-standing controversy on whether membrane lipids or proteins are the target for general anesthetics. The plasma membrane-associated Ca2+-ATPase of synaptosomes has recently been established as a model system for general anesthesia, the protein interior being the proposed target site (M.M. Lopez, D. Kosk-Kosicka, J. Biol. Chem. 270 (1995) 28239-28245). Multiple-site kinetics is now applied as a mechanistic tool to analyze inhibition by organic solvents and general anesthetics. A close fit to the experimental data points was achieved using the complex equations for a competitive displacement of lipid activators from multiple sites on the protein surface. Inhibitor dissociation constants were about 1. 6x105-fold higher than the microscopic lipid dissociation binding constants that are derived here for the first time. Binding of lipid therefore is by -7.1 kcal/mole favored over that of the tested inhibitors. The latter are nevertheless effective because in the model used displacement of only few of the lipid solvation molecules cause complete inhibition. The lipid/protein interface rather than protein or lipid alone appeared to be the anesthetic target site. PMID- 9858724 TI - Novel sequence organization and insertion specificity of IS605 and IS606: chimaeric transposable elements of Helicobacter pylori. AB - IS605, an insertion sequence (IS) that is unusual in containing homologs of genes for the single putative transposases of two other unrelated IS elements (IS200 and IS1341), was found in nearly one-third of a set of 238 independent isolates of the gastric pathogen Helicobacter pylori. Hybridization and PCR tests indicated that any strain carrying one of these ORFs also carried the other, which implies that both ORFs are in the same unit of transposition. The IS605 ends and target sites for insertion were identified by sequencing eight preexisting insertions in strain NCTC11638, corresponding empty sites in other strains, and new transpositions in E. coli of an IS605 derivative marked with a selectable chloramphenicol-resistance gene. These tests showed that IS605 is also unusual in: (1) having unique, not inverted repeat, ends; (2) not duplicating (or deleting) target sequences during transposition; and (3) inserting with its left (IS200-homolog) end next to 5'-TTTAA or 5'-TTTAAC. IS605 was implicated in at least two genome rearrangements in strain NCTC11638. A second member of the IS605 family, called IS606 (25% amino acid identity to IS605 in inferred proteins) was found in one-third of 38 H. pylori strains tested, many of which did not carry IS605. The features of these two chimaeric IS elements are discussed in terms of possible transposition mechanisms, IS element evolution, and effects of IS elements on genome organization and evolution in the microbes that they inhabit. PMID- 9858726 TI - Comparison between the dynamics of lipid/gramicidin A systems in the lamellar and hexagonal phases: a solid-state 13C NMR study. AB - We have investigated the effect of gramicidin A on the dynamics of two model membranes: dimyristoylphosphatidylcholine (DMPC) in the lamellar phase at a lipid to-peptide molar ratio of 10:1 and dioleoylphosphatidylcholine (DOPC) in the hexagonal HII phase at a lipid-to-peptide molar ratio of 5:1. Natural abundance 13C nuclear magnetic resonance (NMR) spectroscopy was used in combination with magic angle spinning to increase the spectral resolution, therefore allowing the different regions of the lipid bilayers to be investigated from the same spectra. 31P NMR was also used to detect and confirm the formation of the DOPC HII phase in the presence of gramicidin A. In order to examine the effect of gramicidin A on both the fast and slow motions of DMPC and DOPC, the 1H spin-lattice relaxation times in the laboratory frame (HT1) as well as the 1H spin-lattice relaxation times in the rotating frame (HT1rho) were calculated for each resolved protonated lipid resonance in the 13C spectra. For both DMPC and DOPC, we found that the presence of gramicidin A does not significantly affect the fast motions of the lipid acyl chains but increases slightly the fast motions of the polar head group. However, the HT1rho are significantly decreased, this effect being more pronounced for DOPC most likely due to a decrease in the rate of the lipid lateral diffusion. PMID- 9858725 TI - Regulated nuclear entry of the C. elegans Pax-6 transcription factor. AB - It is shown that the C. elegans Pax-6 locus encodes two protein isoforms. One contains a Paired DNA binding domain as well as a homeodomain; the other consists only of the carboxy-terminal portion of the locus encoding the homeodomain. These two isoforms are expressed in a variety of postembryonic cell lineages. In one set of lineages, nuclear localization of a homeodomain-only form (MAB-18 isoform) appears to be under temporal and spatial control. Nuclear localization of MAB-18 is correlated with the genetic requirement for mab-18 and with activation of a reporter gene driven by a mab-18 promoter. Reporter gene expression is dependent on mab-18 gene activity. It is hypothesized that a positive feedback loop is activated by regulated nuclear entry. PMID- 9858727 TI - Observations on template switching during DNA replication through long inverted repeats. AB - The bacterial Tn5 transposon consists of a pair of long inverted repeats flanking a central region that carries genes for antibiotic resistance. An analysis of DNA replication through Tn5 by two-dimensional gel electrophoresis has revealed two interesting features: (i) a spike representing X-shaped molecules, and (ii) a spot representing a barrier on the arc of Y-shaped replication intermediates. The electrophoretic behavior of various restriction fragments derived from this region indicates that the X molecules contain two linear Tn5 fragments joined together by a cross connection (Holliday junction). However, their formation is recA independent. The junction seems to connect the right inverted repeat of one fragment to the left inverted repeat of the other. The structure of the X molecules suggests that they could be formed by template switching when synthesis of the leading strand enters the right inverted repeat. One possible mechanism is that switching occurs at the center of a transient cruciform structure. A similar switching event, occurring when synthesis of the leading strand enters the left inverted repeat, could give rise to the barrier. These results imply that the inviability of palindromic DNA, which has previously been attributed to the slowing down of replication, may actually be caused by frequent template switching. PMID- 9858728 TI - The Notch signalling pathway in hair growth. AB - The Notch signalling pathway is an important mediator of cell fate selection whose involvement in epidermal appendage formation is now becoming recognised. Hair follicle development and hair formation involve the co-ordinated differentiation of several different cell types in which Notch appears to have a role. We report intricate expression patterns for the Notch-1 receptor and three ligands, Delta-1, Jagged-1 and Jagged-2 in the hair follicle. Notch-1 is expressed in ectodermal-derived cells of the follicle, in the inner cells of the embryonic placode and the follicle bulb, and in the suprabasal cells of the mature outer root sheath. Delta-1 is only expressed during embryonic follicle development and is exclusive to the mesenchymal cells of the pre-papilla located beneath the follicle placode. Expression of Jagged-1 or Jagged-2 overlaps Notch-1 expression at all stages. In mature follicles, Jagged-1 and Jagged-2 are expressed in complementary patterns in the follicle bulb and outer root sheath, Jagged-1 in suprabasal cells and Jagged-2 predominantly in basal cells. In the follicle bulb, Jagged-2 is localised to the inner (basal) bulb cells next to the dermal papilla which do not express Notch-1, whereas Jagged-1 expression in the upper follicle bulb overlaps Notch-1 expression and correlates with bulb cell differentiation into hair shaft cortical and cuticle keratinocytes. PMID- 9858729 TI - Effect of cholesteryl hemisuccinate on the interfacial properties of phosphatidylcholine bilayers. AB - Cholesteryl hemisuccinate (CHEMS) is an amphipathic lipid that can regulate cell growth. A comparison of the effects of CHEMS and cholesterol on 1-palmitoyl-2 oleoyl-sn-glycero-3-phosphocholine (POPC) and 1,2-dipalmitoyl-sn-glycero-3 phosphocholine (DPPC) bilayers was investigated using fluorescence techniques. In liquid-crystalline phase POPC bilayers, CHEMS increased the interfacial surface charge, but was less effective than cholesterol in reducing acyl chain mobility and interfacial hydration. In liquid-crystalline phase DPPC bilayers, CHEMS and cholesterol were equally effective in reducing acyl chain mobility. Similar to the POPC matrix, CHEMS increased the interfacial surface charge and cholesterol decreased the surface hydration. The different effect of cholesterol and CHEMS on acyl chain mobility may be due to a preferential interaction of cholesterol with POPC. In gel phase DPPC bilayers, CHEMS and a succinylated pyrenyl cholesterol analog exhibited different effects on membrane physical-chemical properties than cholesterol. Succinylation also increased the rate of transfer of the pyrenyl cholesterol analog between single unilamellar vesicles approximately seven fold. This process demonstrated first-order kinetics which indicated that transbilayer migration was not a rate-limiting step. The succinylation of cholesterol places a carboxyl group at the lipid-water interface and the sterol ring deeper in the bilayer. For a structural model to explain its biological properties, CHEMS should be considered a bulky fatty acid. PMID- 9858730 TI - Characterization of two frizzled8 homologues expressed in the embryonic shield and prechordal plate of zebrafish embryos. AB - We have isolated and characterized two complete cDNA clones, Zfz8a and Zfz8b, which encode zebrafish Frizzled (Fz) homologues. The predicted protein sequences, spanning 579 and 576 amino acid residues for ZFz8a and ZFz8b, respectively, were highly homologous (78%) to each other and contained an extracellular cysteine rich domain and seven transmembrane domains that are well conserved in Fz receptor protein members. In comparison with other Fz family members, ZFz8a and ZFz8b showed the highest homology with mouse Fz8 (MFz8), sharing 84 and 76% amino acid identity, respectively. The presence of Zfz8a and Zfz8b transcripts was detected by in situ hybridization in zebrafish embryos from the 512 cell stage, and their appearance in the future dorsal region could be observed before embryos reached the 30% epiboly stage. At shield stage, Zfz8a transcripts were expressed in both epiblast and shield whereas expression of Zfz8b was only detected in the embryonic shield. During gastrula stages, both Zfz8a and Zfz8b transcripts were found in anterior dorsal regions of the involuting mesendoderm (future prechordal plate). By the 2- to 3-somite stage, expression of both Zfz8a and Zfz8b was restricted to the prechordal plate and prospective anterior neurectoderm, although expression of the Zfz8a gene was no longer present in the most anterior portion of the prechordal plate, the polster. In one-eyed pinhead mutant embryos, which lack prechordal plate, both Zfz8a and Zfz8b transcripts were reduced, confirming the prechordal plate specificity of Zfz8a and Zfz8b gene expression. These results provide an additional evidence supporting the role of Wnt signaling in organizer-mediated axial patterning. PMID- 9858731 TI - Regulatory implications of protein assemblies at the gamma origin of plasmid R6K a review. AB - Recognition of the replication origin (ori) by initiator protein is a recurring theme for the regulated initiation of DNA replication in diverse biological systems. The objective of the work reviewed here is to understand the initiation process focusing specifically on the gamma-ori of the antibiotic-resistance plasmid R6K. The control of gamma-ori copy number is determined by both plasmid encoded and host-encoded factors. The two central regulatory elements of the plasmid are a multifunctional initiator protein pi, and sequence-related DNA target sites, the inverted half-repeats (IRs) and the direct repeats (DRs). The replication activator and inhibitor activities of pi seem to be at least partially distributed between two naturally occurring pi polypeptides (designated by their molecular weights pi35.0 and pi30.5). Regulatory variants of pi with altered states of oligomerization in nucleoprotein complexes with DRs and IRs have been isolated. The properties of these mutants laid the foundation for our model of pi protein activity which proposes that different protein surfaces are required for the formation of functionally distinct complexes of pi with DRs and IRs. These mutants also suggest that pi polypeptides have a modular structure; the C-terminus contains the DNA-binding domain while the N-terminus controls protein oligomerization. Additionally, pi35.0 binds to a novel DNA sequence in the A+T-rich segment of gamma-ori. This binding site is at or near the site from which synthesis of the leading strand begins. PMID- 9858733 TI - The phospholipid-deficient pho1 mutant of Arabidopsis thaliana is affected in the organization, but not in the light acclimation, of the thylakoid membrane. AB - The pho1 mutant of Arabidopsis has been shown to respond to the phosphate deficiency in the leaves by decreasing the amount of phosphatidylglycerol (PG). PG is thought to be of crucial importance for the organization and function of the thylakoid membrane. This prompted us to ask what the consequences of the PG deficiency may be in the pho1 mutant when grown under low or high light. While in the wild-type, the lipid pattern was almost insensitive to changes in the growth light, PG was reduced to 45% under low light in the mutant, and it decreased further to 35% under high light. Concomitantly, sulfoquinovosyl diacylglycerol (SQDG) and to a lesser extent digalactosyl diacylglycerol (DGDG) increased. The SQDG increase correlated with increased amounts of the SQD1 protein, an indicator for an actively mediated process. Despite of alterations in the ultrastructure, mutant thylakoids showed virtually no effects on photosynthetic electron transfer, O2 evolution and excitation energy allocation to the reaction centers. Our results support the idea that PG deficiency can at least partially be compensated for by the anionic lipid SQDG and the not charged lipid DGDG. This seems to be an important strategy to maintain an optimal thylakoid lipid milieu for vital processes, such as photosynthesis, under a restricted phosphate availability. PMID- 9858732 TI - Identification and developmental expression of Ci-isl, a homologue of vertebrate islet genes, in the ascidian Ciona intestinalis. AB - Here we describe the cloning and expression pattern of Ci-isl, a homologue of vertebrate genes, in the ascidian. Early in development, Ci-isl expression occurs in the primordia of palps and brain vesicle, then in the tailbud embryo it is transiently extended to the notochord cells. At larva stage, the expression is down-regulated in the notochord, and it persists predominantly in the compartments of the nervous system. These observations indicate that also in invertebrates, islet genes show an expression pattern during differentiation of the nervous system. PMID- 9858734 TI - TrfA dimers play a role in copy-number control of RK2 replication. AB - Copy-number regulation of the broad-host-range plasmid RK2 is dependent on the plasmid-encoded initiator protein, TrfA, and the RK2 origin of replication. The handcuffing model for copy-number control proposes that TrfA-bound oris reversibly couple to prevent the further initiation of plasmid replication when the copy number in vivo is at or above the replicon-specific copy number. TrfA mutants have been isolated which allow for oriV replication at elevated copy numbers. To better understand the mechanism of 'handcuffing', the copy-up TrfA(G254D/S267L) mutant was characterized further. In the present study we show by size exclusion chromatography and native gel electrophoresis that unlike wt TrfA which is largely dimeric, purified His6-TrfA(G254D/S267L) is primarily monomeric. In vivo, TrfA33(G254D/S267L) supports replication of an RK2 ori plasmid in trans at a greatly elevated copy number, while in cis the plasmid exhibits runaway replication. However, expression of either of two previously isolated DNA-binding defective TrfA mutants, TrfA33(P151S) or TrfA33(S257F), in a cell transformed with a mini-RK2 replicon encoding TrfA33(G254D/S267L) results in suppression of the runaway phenotype. His6-TrfA(P151S) and His6-TrfA(S257F) purify as dimers, and when expressed in vivo are incapable of supporting RK2 plasmid replication. In contrast, combination of the trfA(P151S) or trfA(S257F) mutation with the trfA(G254D/S267L) mutations results in the expression of mutant TrfA proteins which are mainly monomers and which can no longer restore copy control to replication directed by TrfA33(G254D/S267L) in vivo. On the basis of these findings a handcuffing model is proposed, whereby oriV-bound TrfA monomers are coupled by dimeric TrfA molecules. PMID- 9858736 TI - pBRINT-Ts: a plasmid family with a temperature-sensitive replicon, designed for chromosomal integration into the lacZ gene of Escherichia coli. AB - A pBRINT-Ts family of integrative vectors for Escherichia coli was constructed by using a temperature-sensitive replicon derived from pSC101, a region of homology to the lacZ gene, and various antibiotic resistance markers (kanamycin, chloramphenicol and gentamycin) for selection of the integrants. The gene or group of genes to be integrated can be inserted into a multiple cloning site, flanked by an antibiotic resistance marker and lacZ sequences. A simple and rapid procedure was developed for the selection of cells where allelic exchange had occurred. With this procedure, the efficiency of integration of around 10-3 was observed, using several E. coli strains. From colonies with an integrated pBRINT Ts plasmid, we detected an average allelic exchange event frequency of 7.5%. As a test for this system, we integrated the amy gene that codes for the alpha-amylase from B. stearothermophilus, into the lacZ gene of E. coli W3110. Production of alpha-amylase was found to be proportional to copy number; at up to 10 copies per chromosome. PMID- 9858735 TI - Cloning and characterization of Sel-1l, a murine homolog of the C. elegans sel-1 gene. AB - The Notch signaling pathway regulates specification and proliferation in a variety of cell lineages in invertebrates and vertebrates. We have cloned a murine homolog of SEL-1, a key negative regulator of the Notch pathway in Caenorhabditis elegans. Murine SEL-1L (mSEL-1L) protein exhibits a high degree of similarity to SEL-1, including a signal peptide and the C-terminal region required for SEL-1 function in C. elegans. This mammalian homolog of sel-1 is widely expressed in adult mouse and human tissues, with particularly high levels in the pancreas. RNA in situ analysis of developing mouse embryos indicates that mSEL-1L is moderately expressed throughout the neural tube and dorsal root ganglia, with particularly high levels in the floor plate of the neural tube beginning at E10.5 and increasing at E11.5. Expression is high at E14.5 and E17.5 in the acini of the pancreas, and moderate in the epithelial cells of the gut villi. We localized the SEL-1L protein to the cytosol, possibly in intracellular vesicles, in a beta-islet-derived tumor cell line (RinM). PMID- 9858737 TI - Destabilization of cationic lipid vesicles by an anionic hydrophobically modified poly(N-isopropylacrylamide) copolymer: a solid-state 31P NMR and 2H NMR study. AB - The effect of binding PNIPAM-Py-Gly, a copolymer of N-isopropylacrylamide, N-[4 (1-pyrenyl)butyl]-N-n-octadecylacrylamide and N-glycydyl-acrylamide, on membrane stability in cationic multilamellar vesicles (MLVs) was examined using solid state phosphorus (31P) and deuterium (2H) nuclear magnetic resonance (NMR) spectroscopy. For MLVs of composition n-octadecyldiethylene oxide (ODEO)+cholesterol (CHOL)+1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)+dimethyldioctadecylammonium bromide (DODAB) (molar ratios 75:10.5:10.5:4), PNIPAM-Py-Gly induced a complete conversion from a bilayer-type 31P NMR spectrum to one characteristic of lipids undergoing isotropic motional averaging, indicating the existence of regions of high local membrane curvature. This response was sustained even at elevated temperatures. For MLVs of composition POPC+1,2-dioleoyloxy-3-(trimethylammonio)-propane (DOTAP), only at high levels of DOTAP and ionic strength did PNIPAM-Py-Gly induce even a partial conversion to an isotropic-type 31P NMR spectrum. At lower pH this effect was diminished. Raising the temperature eliminated the isotropic 31P NMR spectral component, and this effect was not reversible upon returning to room temperature. 2H NMR spectroscopy of headgroup-deuterated DOTAP and POPC confirmed the 31P NMR results, but did not provide specific surface electrostatic information. We conclude that the binding of PNIPAM-Py-Gly to phospholipid-based vesicles is dominated by electrostatic attraction between cationic lipids and the polymer's glycine residues. At high binding levels, the polymer assumes a collapsed conformation at the surface, resulting in regions of high local curvature of the lipid assembly. For ODEO based liposomes, these effects are magnified by the additional contribution of hydrogen bonding to the strength of polymer binding. PMID- 9858738 TI - Using chromosomal lacIQ1 to control expression of genes on high-copy-number plasmids in Escherichia coli. AB - Transcription of the lac and the hybrid tac promoters is repressed by the lac repressor and induced by the non-metabolizable substrate IPTG. The degree of repression depends upon the ratio of LacI molecules in a cell to the DNA operator sites. In the absence of an inducer, repression of Ptac on a high-copy-number (hcn) plasmid was equivalent in strains containing lacIQ1 on the chromosome, or lacI+ on the plasmid, but not from strains with lacI+ or lacIQ only on the chromosome. Induction of Ptac on hcn plasmids in strains in which expression was controlled by lacIQ1 occurred at very low inducer concentrations (3-10microM IPTG) and reached levels significantly higher than in strains with lacI+ on the plasmid. Greater than 300-fold induction of a beta-LacZ fusion was observed, and >600-fold induction was estimated from recombinant hemoglobin synthesis. Transcription from PlacIQ1 initiated in the same point as PlacI+, but was 170 fold stronger, consistent with the lac repressor levels required to control LacI regulated genes on hcn plasmids. The DNA sequence upstream of lacI was used to develop a simple PCR test to identify lacIQ1 by a characteristic 15-bp deletion. This deletion created a consensus -35 hexamer, responsible for the increased lacI transcription, and was easily detectable in a variety of strains. Using lacIQ1 hosts eliminates the requirement to maintain lacI on the plasmid to regulate gene expression on hcn expression plasmids. PMID- 9858739 TI - GalR-mediated repression and activation of hybrid lacUV5 promoter: differential contacts with RNA polymerase. AB - The GalR repressor regulates expression of genes of the gal regulon in Escherichia coli. We studied the regulatory effect of GalR in vitro on a heterologous promoter, lacUV5, by placing the GalR-binding site, OE, at different locations upstream of this promoter. Despite the fact that the lacUV5 promoter is transcribed efficiently by RNA polymerase (RNP) alone, GalR modulated transcription from many of the PlacUV5 variants. Depending on the location of OE and the neighboring DNA sequence, GalR repressed, activated or had no effect on the promoter. Both repression and activation involved formation of GalR-RNP-DNA ternary complexes and required an intact c-domain of the alpha subunit of the holoenzyme. These results support the differential contact model of a regulator action, in which a regulator differentially binds to, and lowers the energy of, intermediates of transcription initiation either to hinder or to facilitate a step of initiation. The nature of the contacts depends upon the context, i.e. the geometry of the ternary complex. The observed repression and activation effect of GalR on a heterologous promoter also underscores the point that a regulator is not a dedicated protein for repression or for activation. PMID- 9858740 TI - Structural and functional evolution of the basal ganglia in vertebrates. AB - While a basal ganglia with striatal and pallidal subdivisions is 1 clearly present in many extant anamniote species, this basal ganglia is cell sparse and receives only a relatively modest tegmental dopaminergic input and little if any cortical input. The major basal ganglia influence on motor functions in anamniotes appears to be exerted via output circuits to the tectum. In contrast, in modern mammals, birds, and reptiles (i.e., modern amniotes), the striatal and pallidal parts of the basal ganglia are very neuron-rich, both consist of the same basic populations of neurons in all amniotes, and the striatum receives abundant tegmental dopaminergic and cortical input. The functional circuitry of the basal ganglia also seems very similar in all amniotes, since the major basal ganglia influences on motor functions appear to be exerted via output circuits to both cerebral cortex and tectum in sauropsids (i.e., birds and reptiles) and mammals. The basal ganglia, output circuits to the cortex, however, appear to be considerably more developed in mammals than in birds and reptiles. The basal ganglia, thus, appears to have undergone a major elaboration during the evolutionary transition from amphibians to reptiles. This elaboration may have enabled amniotes to learn and/or execute a more sophisticated repertoire of behaviors and movements, and this ability may have been an important element of the successful adaptation of amniotes to a fully terrestrial habitat. The mammalian lineage appears, however, to have diverged somewhat from the sauropsid lineage with respect to the emergence of the cerebral cortex as the major target of the basal ganglia circuitry devoted to executing the basal ganglia-mediated control of movement. PMID- 9858741 TI - Cecropins induce the hyperosmotic stress response in Escherichia coli. AB - Cecropin A and B, below or near their minimum inhibitory concentrations in viable Escherichia coli, interfered with the rapid NaCl-induced hyperosmotic shrinkage of the cytoplasmic volume (plasmolysis), and also activated the promoter of the hyperosmotic stress gene osmY. The same promoter was also expressed by hyperosmolar NaCl or sucrose, two of the most commonly used antimicrobial food preservatives. Stress responses were monitored during the logarithmic growth phase of E. coli strains that contain specific promoters fused to a luxCDABE operon on a plasmid. The luminescence assay, developed to monitor the transcriptional response to stresses, is based on the premise that organisms often respond and adapt to sublethal environmental adversities by increased expression of stress proteins to restore homeostasis. The luminescence response from these fusion strains to a specific stress occurs as the transcription at the promoter site is activated. Cecropins induced luminescence response only from the osmY-luxCDABE fusion, but not the corresponding stress promoter activation associated with macromolecular or oxidative damage, or leakage of the cytoplasmic content including the proton gradient. The inhibitory effect of cecropins on plasmolysis is interpreted to suggest that the primary locus of action of these antimicrobial peptides in the periplasmic space is on the coupling between the inner and outer membrane. PMID- 9858742 TI - Effect of mutation of glycosylation sites on the Na+ dependence of steady-state and transient currents generated by the neuronal GABA transporter. AB - The GABA (gamma-aminobutyric acid) transporter (GAT1) belongs to a superfamily of secondary active uptake systems for neurotransmitters that depend on the electrochemical gradients for Na+ and Cl-. In the GAT1, two Na+ ions and one Cl- ion are co-transported with one GABA molecule. Steady-state transport activity and transient charge movements during partial reactions of the transport cycle of the GAT1 of mouse brain expressed in Xenopus oocytes were investigated by two electrode voltage clamp. Functional expression was demonstrated by Na+-dependent [3H]GABA uptake. Effects of mutation of two out of three N-glycosylation sites located in the extracellular loop between transmembrane domains 3 and 4 (Asn176, Asn181, Asn184) were analysed. Simultaneous substitution of two Asn by Asp leaves the transport system intact but leads to a reduction in turnover and complex changes in the interaction of external Na+ with the transport protein. If Asn176 is mutated to Asp and simultaneously Asn181 to Gly, no transport and no charge movements can be detected. In conclusion, mutations of the glycosylation sites result in altered transport, and the local conformation at Asn181 is critical for expression of transport activity. PMID- 9858743 TI - Recognition of binding sites I and II by the TrpI activator protein of pseudomonas aeruginosa: efficient binding to both sites requires InGP even when site II is replaced by site I. AB - TrpI protein, the activator of transcription of the trpBA operon of three species of fluorescent Pseudomonads, bends the DNA when it forms either of two well characterized complexes with the trpBA regulatory region. In complex 1, TrpI is bound only to its strong binding site (site I), whereas in complex 2, which is required for activation of the trpBA promoter, TrpI is bound both to site I and to the weaker site II. Indoleglycerol phosphate (InGP) strongly stimulates formation of complex 2 and is required for activation. The present study focuses on the binding of TrpI to DNA containing a duplication of site I and the effect of the duplication on TrpI-induced DNA bending. We find that even on DNA containing a tandem (direct or inverted) duplication of site I, the formation of DNA-TrpI complexes with both sites occupied is strongly stimulated by InGP. Thus, even when TrpI binding to two adjacent sites needs not be cooperative, InGP significantly promotes the formation of complex 2. Gel binding data indicate that InGP can have several effects: (1) TrpI molecules bound to either of two adjacent strong binding sites appear to interfere with binding to the other site; InGP relieves this apparent interference. (2) InGP increases the intrinsic affinity of TrpI for sites I and II and/or enhances cooperative TrpI binding to adjacent DNA sites. Furthermore, a third molecule of TrpI can form a footprint adjacent to the duplication on DNA containing a direct (but not inverted) repeat of site I, indicating that TrpI bound to site I is oriented asymmetrically in spite of the quasi-symmetry of the binding site. The calculated bending angle for DNA in complex 2 is increased by approximately 20 degrees when site I is substituted in either orientation for site II; thus, on DNA containing a site I duplication, the bending angle of complex 2 is nearly twice that of complex 1. PMID- 9858744 TI - Antiamoebin can function as a carrier or as a pore-forming peptaibol. AB - Antiamoebin is a 16-residue polypeptide whose crystal structure and lytic activity in membrane vesicles have recently been reported. It is a bent helical molecule and a member of the peptaibol family of antibiotics. Under conditions which produce voltage-dependent conductance activity by other members of the family, no single-channel conductance was detected for antiamoebin, and a carrier like mechanism was put forward to account for its mode of action. We now present evidence for pore formation that is largely voltage-insensitive, with large amplitude single-channel events on top of a background conductance that may account for the previously proposed carrier-like activity. Thus, antiamoebin may be the first instance of a peptide which can function both as an ion carrier and a pore former. PMID- 9858746 TI - Functional expression of sperm angiotensin II type I receptor in Xenopus oocyte: modulation of a sperm Ca2+-activated K+ channel. AB - In addition to Ca2+ and K+ fluxes, angiotensin II (Ang II) has been shown to influence sperm motility. The present study investigated the involvement of angiotensin II type 1 receptor (AT1) in mediating the modulatory effect of Ang II on a sperm Ca2+-activated K+ channel expressed in Xenopus oocytes injected with RNAs of spermatogenic cells. Ang II at a concentration of 1 microM was found to potentiate the ionomycin-induced current, previously demonstrated to be mediated by a 'Maxi' Ca2+-activated K+ channel. However, at higher concentration, 20 microM, Ang II was found to suppress the ionomycin-induced current. Both potentiating and inhibitory effects of Ang II were blocked by losartan, a specific antagonist of AT1 receptors. Immunohistochemical studies further confirmed the presence of AT1 receptors in spermatogenic cells while expression of AT1 receptor mRNA was demonstrated by RT-PCR. These results suggest that Ang II may influence sperm motility as well as other sperm function by acting on AT1 receptors, and exerting potentiating and inhibitory effects on the Ca2+-activated K+ channels. PMID- 9858745 TI - Transcriptional activation of the catechol and chlorocatechol operons: variations on a theme. AB - The ortho-cleavage pathways of catechol and 3-chlorocatechol are central catabolic pathways of Pseudomonas putida that convert aromatic and chloroaromatic compounds to tricarboxylic acid (TCA)-cycle intermediates. They are encoded by the evolutionarily related catBCA and clcABD operons, respectively. Expression of the cat and clc operons requires the LysR-type transcriptional activators CatR and ClcR, and the inducer molecules cis,cis-muconate and 2-chloro-cis,cis muconate. In addition to sequence similarities, CatR and ClcR share functional similarities which allow catR to complement clcR mutants. DNase-I footprinting, DNA bending and in vitro transcription analyses with RNA polymerase mutants indicate that CatR and ClcR activate transcription via a similar mechanism which involves interaction with the C-terminal domain of the alpha-subunit (alpha-CTD) of RNA polymerase. In vitro transcription assays with different regions of the clc promoter indicate that the ClcR dimer bound to the promoter proximal site (the activation binding site) interacts with the alpha-CTD. Gel shift assays and DNase-I footprinting have demonstrated that CatR occupies two adjacent sites proximal to the catBCA promoter in the presence of inducer and an additional binding site within the catB structural gene called the internal binding site (IBS). CatR binds the IBS with low intrinsic affinity that is increased by cooperativity in presence of the two promoter binding sites. Site-directed mutations in the IBS indicate a probable cis-acting repressor function for the IBS. The location of the IBS within the catB structural gene, the cooperativity observed in footprinting studies and phasing studies suggest that the IBS participates in the interaction of CatR with the upstream binding sites by looping out the intervening DNA. Although the core transcriptional activation mechanisms of CatR and ClcR have been conserved, nature has provided some flexibility to respond to different environmental signals in addition to the presence of inducer. Transcriptional fusion studies demonstrate that the expression from the clc promoter is repressed when the cells are grown on succinate, citrate or fumarate and that this repression is ClcR-dependent and occurs at the transcriptional level. The presence of these organic acids did not affect the expression from the cat promoter. In vitro transcription assays demonstrate that the TCA-cycle intermediate, fumarate, directly and specifically inhibits the formation of the clcA transcript. No such inhibition was observed when CatR was used as activator on either the cat or clc template. Since both the catechol and the chlorocatechol pathways feed into the TCA cycle, but only the chlorocatechol pathway is inhibited by fumarate, there is a subtle difference in the regulation of these two pathways where intracellular sensing of a TCA-cycle intermediate leads to a reduction of chloroaromatic degradation. PMID- 9858747 TI - Sequence of a pyrimidine-selective Na+/nucleoside cotransporter from pig kidney, pkCNT1. AB - A 3 kb cDNA called pkCNT1, a new member of the Na+/nucleoside cotransporter family, was cloned from pig kidney and sequenced. The sequence of pkCNT1 encodes a 647 amino acid protein that is 84% identical to the sequence of the rat pyrimidine-selective Na+/nucleoside cotransporter, rCNT1. pkCNT1 transports pyrimidines, such as thymidine and uridine, and has a Km for uridine of 9 microM. PMID- 9858748 TI - Genetic organization of the regions associated with surface polysaccharide synthesis in Vibrio cholerae O1, O139 and Vibrio anguillarum O1 and O2: a review. AB - Vibrio cholerae and V. anguillarum are recognized as aquatic-borne human and fish pathogens, respectively. Based upon analyses of several genes and the presence of novel genetic elements it seems that these two species are very closely related. Studies in this laboratory have identified an association of IS1358 with rfb and capsule loci in these two species. The most recent findings suggest that IS1358 is associated with the rfb region in V. cholerae O1 and O139 and in V. anguillarum O1 and O2. In addition, the rfb region in both V. cholerae serogroups and in V. anguillarum O1 is limited at one end by gmhD. These features make it feasible to envisage a mechanism by which the evolution of new rfb genes is taking place involving IS1358 and the region around gmhD. Furthermore, it is possible to envisage that there is or has been an exchange of genetic material between these species leading to new rfb/capsule regions. This review examines the genetics and biosynthesis of the O-antigen and capsule of V. cholerae O1 and O139, as well as the V. anguillarum serogroup O1 and the role of IS1358. Throughout this review we have used the new nomenclature for rfb genes proposed by. PMID- 9858749 TI - Mapping of 41 chemotaxis, flagellar and motility genes to a single region of the Sinorhizobium meliloti chromosome. AB - Three previously identified gene clusters that contain chemotaxis (che), flagellar (fla) and motility (mot) genes of Sinorhizobium meliloti (formerly Rhizobium meliloti) were mapped to a contiguous 45-kb region of the S. meliloti RU11/001 genome by pulsed-field gel electrophoresis (PFGE) in combination with Southern hybridization. The entire region was cloned and sequenced. The map combines 32 che, fla (flg, flh, fli) and mot genes and nine new open reading frames that probably encode taxis-related functions as well. It is concluded that between 80 and 90% of the genes responsible for chemotaxis and motility are located in a single region of the S. meliloti chromosome near the his-39 marker. PMID- 9858750 TI - A fluorescence study of ligand-induced conformational changes in cytosolic fructose-1,6-bisphosphatase from germinating castor oil seeds. AB - The intrinsic fluorescence of homogeneous castor oil seed cytosolic fructose-1,6 bisphosphatase (FBPasec) was used as an indicator of conformational changes due to ligand binding. Binding of the substrate and the inhibitor fructose-2,6 bisphosphate (F-2,6-P2) was quantitatively compared to their respective kinetic effects on enzymatic activity. There are two distinct types of substrate interaction with FBPasec, corresponding to catalytic and inhibitory binding, respectively. Inhibitory substrate binding shares several characteristics with F 2,6-P2 binding which indicates that both ligands bind at the same site. However, F-2,6-P2 does not prevent fluorescence transitions attributed to catalytic substrate binding. The marked synergistic inhibition of FBPasec by AMP and F-2,6 P2 appears to arise via AMP's promotion of F-2,6-P2 binding. Based on the X-ray crystal structure of porcine kidney FBPase our modelling studies suggest the existence of a distinct F-1,6-P2/F-2,6-P2 inhibitory binding site which partially overlaps with the enzyme's catalytic site. We propose that a pronounced allosteric transition mediated by AMP binding increases access of F-1,6-P2 and F 2,6-P2 to this common inhibitory binding site. PMID- 9858751 TI - The organization of corticothalamic projections: reciprocity versus parity. AB - All neocortical areas receive inputs from and project back to the thalamus. It is often said that the corticothalamic projections are organized in a way that reciprocates the spatial distribution of thalamocortical pathways. The present review examines to what extent this rule of reciprocity is actually supported by the most recent neuroanatomical data, particularly those relating to the central organization of the vibrissal sensory system in the rat. A critical survey of previous studies is made and new results are presented concerning the fine grained organization of corticothalamic projections in this sensory system. Together, prior results and the present set of new data confirm the existence of both, reciprocal and nonreciprocal patterns of corticothalamic connectivity. This conclusion leads us to propose that the spatial organization of corticothalamic connections complies with a more fundamental rule, the rule of parity, from which reciprocity follows as a general, but not obligatory consequence. The rule of parity states that the distribution of corticothalamic projections across and within the thalamic nuclei is determined by the branching patterns of the different classes of prethalamic afferents. The anatomical, developmental and physiological consequences of this rule are discussed. The rule of parity suggests that, according to the behavioral context, both prethalamic and corticothalamic pathways may function in a feedback mode. PMID- 9858752 TI - Two intertwined methylation activities of the MmeI restriction-modification class IIS system from Methylophilus methylotrophus. AB - The class-IIS restriction endonuclease, R.MmeI, was isolated from Methylophilus methylotrophus. It was originally described as a monomeric enzyme, with the native Mr 105000+/-7000, which did not cleave DNA efficiently [Boyd et al. (1986) Nucleic Acids Res. 14, 5255-5274; Tucholski et al. (1995) Gene 157, 87-92]. However, it was discovered that R.MmeI endonucleolytic activity is enhanced by S adenosyl-l-methionine (AdoMet) and sinefungin, an analogue of AdoMet. Surprisingly, the purified R.MmeI endonuclease was found to have a second enzymatic activity, namely methylation of the adenine residue to N6-methyladenine in the top strand of the MmeI-recognition sequence, 5'-TCCR*AC-3' (*A=meA. The R.MmeI methylating activity requires AdoMet and is increased in the presence of several divalent cations, 20-fold by Mg2+ or Ca2+, and less by Mn2+, Zn2+ and Co2+; however, methylation is inhibited entirely by sinefungin, at concentrations above 9microM. The latter observation shows that the enhancing effect of AdoMet or sinefungin on the DNA cleavage was not related to the process of DNA methylation. Furthermore, a second component of the MmeI restriction-modification system, a M.MmeI methyltransferase, was isolated and purified. The M.MmeI protein was found to have an Mr of 48000+/-2000 (under denaturing conditions) and to methylate both adenine residues (*A) in the MmeI-recognition sequence 5'-TCCR*AC 3'/3'-*AGGYTG-5'. Methylation of the top strand does not inhibit the DNA cleavage by R.MmeI, whereas methylation of both DNA strands blocks the cleavage process. PMID- 9858753 TI - Identification of a beta-lactoglobulin lactosylation site. AB - Thermal treatment of milk leads to non-enzymatic glycosylation of proteins through Maillard reaction. Free NH2 groups of basic amino acids react with the reducing carbonyl group of lactose forming the so-called Amadori products. Electrospray mass spectrometry analysis shows that beta-lactoglobulin (beta-LG), the major whey protein, undergoes lactosylation under industrial thermal treatment. In order to investigate the specificity of reactive sites for lactose binding the analysis of trypsin hydrolysates of beta-LG isolated from different industrial milks was performed. Results demonstrate that Lys-100 is a preferential lactosylation site of beta-LG during industrial milk treatment. These results were confirmed by an analysis of the three-dimensional model of the protein which showed that Lys-100 had the highest solvent accessibility and proximity to another amino group making Lys-100 the best candidate to lactosylation. Lys-47, previously identified by other authors, showed a good proximity to another Lys residue, but an intermediate level of exposition to solvent. PMID- 9858754 TI - A conserved TN8TCCT motif in the octapeptide-encoding region of Pax genes which has the potential to direct cytosine methylation. AB - Our previous findings have shown that the developmental genes Pax7 and Pax3 are differentially methylated; the gene region that encodes the paired domain is hypomethylated, whereas the region that encodes the homeodomain is hypermethylated. For this reason, the known DNA sequence between the paired and homeoboxes was analysed for the presence of a conserved DNA motif to which a modifying protein could bind in order to direct the methylation or demethylation of surrounding gene sequences. The octapeptide-encoding region was found to contain several nucleotides that were highly conserved throughout the Pax gene family from phylogenetically distant species. The most conserved nucleotides are thought to comprise a motif TN8TCCT where N8=any combination of eight nucleotides. A conserved octapeptide-like-encoding sequence containing the TN8TCCT motif was also found in non-Pax genes of higher eukaryotes and in the non coding strand of plants. Moreover, differential methylation seems to be associated with the presence of the TN8TCCT motif in p53 and the human oestrogen receptor genes. The presence of the TN8TCCT motif within an octapeptide-like encoding sequence in human T-cell leukaemia virus type 1 might suggest that the putative recognition motif may have been introduced into various host genomes via some form of retroviral agent. PMID- 9858755 TI - Isolation and characterization of an antifreeze protein from the longhorn sculpin, Myoxocephalus octodecimspinosis. AB - A new type of antifreeze protein was isolated from the serum of the longhorn sculpin, Myoxocephalus octodecimspinosis, by gel filtration and high-performance liquid chromatography. This protein (LS-12) exhibits freezing point depression activity (thermal hysteresis) and ice crystal modification properties similar to those seen for other types of fish antifreeze polypeptide, except that ice crystals grow as hexagonal trapezohedra in the presence of LS-12, rather than hexagonal bipyramids usually seen. Ice crystal etching studies demonstrate that LS-12 does not bind to the hexagonal bipyramidal or secondary prism surfaces reported for the antifreeze polypeptides from winter flounder and shorthorn sculpin, respectively. Circular dichroism studies indicate that LS-12 has an alpha-helix content of about 60% at 1 degreesC, which is in good agreement with a value of about 70% predicted from the amino acid sequence. Limited proteolysis studies and further analysis of the amino acid sequence suggest that LS-12 consists of four amphipathic alpha-helices of similar length which are folded into a four-helix bundle. Based on its size (Mr=12299) and predicted tertiary structure, LS-12 can be regarded as the first example of a new class (type IV) of fish antifreeze protein. PMID- 9858756 TI - What is the role of dopamine in reward: hedonic impact, reward learning, or incentive salience? AB - What roles do mesolimbic and neostriatal dopamine systems play in reward? Do they mediate the hedonic impact of rewarding stimuli? Do they mediate hedonic reward learning and associative prediction? Our review of the literature, together with results of a new study of residual reward capacity after dopamine depletion, indicates the answer to both questions is 'no'. Rather, dopamine systems may mediate the incentive salience of rewards, modulating their motivational value in a manner separable from hedonia and reward learning. In a study of the consequences of dopamine loss, rats were depleted of dopamine in the nucleus accumbens and neostriatum by up to 99% using 6-hydroxydopamine. In a series of experiments, we applied the 'taste reactivity' measure of affective reactions (gapes, etc.) to assess the capacity of dopamine-depleted rats for: 1) normal affect (hedonic and aversive reactions), 2) modulation of hedonic affect by associative learning (taste aversion conditioning), and 3) hedonic enhancement of affect by non-dopaminergic pharmacological manipulation of palatability (benzodiazepine administration). We found normal hedonic reaction patterns to sucrose vs. quinine, normal learning of new hedonic stimulus values (a change in palatability based on predictive relations), and normal pharmacological hedonic enhancement of palatability. We discuss these results in the context of hypotheses and data concerning the role of dopamine in reward. We review neurochemical, electrophysiological, and other behavioral evidence. We conclude that dopamine systems are not needed either to mediate the hedonic pleasure of reinforcers or to mediate predictive associations involved in hedonic reward learning. We conclude instead that dopamine may be more important to incentive salience attributions to the neural representations of reward-related stimuli. Incentive salience, we suggest, is a distinct component of motivation and reward. In other words, dopamine systems are necessary for 'wanting' incentives, but not for 'liking' them or for learning new 'likes' and 'dislikes'. PMID- 9858758 TI - DNA binding properties and processive proofreading of herpes simplex virus type 1 DNA polymerase. AB - The DNA binding properties of herpes simplex virus type 1 DNA polymerase (HSV pol), an alpha-like DNA polymerase, were investigated using an optimized band shift assay. With linear double-stranded DNA (dsDNA), HSV pol formed two complexes. The favored DNA template was dsDNA with protruding 5'-phosphoryl termini. Stable binding of HSV pol was observed with a DNA hairpin containing a primer region of 9 bp of dsDNA, a 6-base loop and a 12-base 5'-terminal single stranded extension. For the polymerization activity of HSV pol on poly(dT) an optimal primer length of 8 to 10 nucleotides was determined. The DNA binding event could be clearly separated from the enzymatic activities by its unique response to divalent cations and salt. Under ionic strength conditions where HSV pol exerts optimal polymerization activity in vitro, novel polymerase-DNA complexes were detected by band-shift analysis. These new complexes were similar while either in DNA polymerase or 3',5' exonuclease mode. Using a polymerase trap method and high-resolution polyacrylamide gel electrophoresis, HSV pol demonstrated internal switching from 3',5' exonuclease to polymerase-active mode during one DNA binding event. These results support the role of HSV pol as a true replicase, which proofreads without dissociating from the DNA template. PMID- 9858757 TI - Characterization of rice endo-beta-glucanase genes (Gns2-Gns14) defines a new subgroup within the gene family. AB - Thirteen new beta-glucanase-encoding genes have been identified in the rice genome. These genes, together with other monocot beta-glucanases, have now been classified into four subfamilies based on the structure and function of the genes. Two tandem gene clusters, Gns2-Gns3-Gns4 and Gns5-Gns6, were classified in the defense-related Subfamily A. Growth-related 1,3;1,4-beta-glucanase Gns1 was classified in Subfamily B. Gns7 and Gns8, together with the barley genes GVI and Hv34, represent Subfamily C. Gns9 and a beta-glucanase gene from wheat were grouped in Subfamily D. Genes in Subfamilies C and D have structures that are distinct from those of the other subfamilies, but there are very little data available on the biochemical or physiological roles of these genes. Gene expression in growing tissues and lack of gene induction in response to disease related treatments suggest that Subfamilies C and D may function in control of plant growth. PMID- 9858759 TI - Characterization of the maize prolamin box-binding factor-1 (PBF-1) and its role in the developmental regulation of the zein multigene family. AB - A maize prolamin box (P-box)-binding factor (PBF-1) has been purified and characterized from immature endosperm tissue. PBF-1 has a molecular weight of 38kDa. It is detected only in endosperm, but not in root or leaf tissues, consistent with its tissue-specific function. Site-directed mutagenesis experiments reveal that both the P-box and its flanking sequences are important for PBF-1 DNA binding. Developmental studies show that PBF-1 accumulates in the endosperm from 8 to at least 30days after pollination (DAP). From 16 to 24DAP, however, multiple shifted bands of protein(s)-DNA complexes can be observed, which correlate with an increase in zein gene expression. PBF-1 can also bind to the P-box from '22-kDa' and '19-kDa' zein promoters, but at a lower affinity than to the '27-kDa' zein promoter. The effects of protein dephosphorylation and zinc ion chelators on PBF-1 DNA binding activity are also shown. A model is proposed where PBF-1 serves as a 'recruiter' of class-specific transcription factors like Opaque2 (O2). PMID- 9858760 TI - Comparison of conformational changes and inactivation of soybean lipoxygenase-1 during urea denaturation. AB - The unfolding and inactivation of soybean lipoxygenase-1 during urea denaturation has been compared. Equilibrium study indicates that inactivation of the enzyme occurs at low urea concentrations before significant conformational change of the molecule as a whole. In the presence of 6.0 M urea, the unfolding of soybean lipoxygenase-1, as monitored by fluorescence intensity, is a triphasic process, while the inactivation of the enzyme shows single-phase kinetics. The rate constant of inactivation is consistent with that of the fast conformational change of the enzyme. The results suggest that active sites of lipoxygenase-1 containing iron cofactor are situated in a limited region of the enzyme molecule that is more fragile to denaturants than the protein as a whole. The kinetic theory of substrate reactions catalyzed by unstable enzymes (Duggleby (1986) J. Theor. Biol. 123, 67-80) has been applied to study the effect of substrate on enzyme inactivation. On the basis of the kinetic equation of substrate reaction in the presence of urea, inactivation rate constants for the free enzyme and enzyme-substrate complex have been determined. The substrate, linoleic acid, has no effect on inactivation of the ferric form of lipoxygenase-1. PMID- 9858761 TI - Modulation of gene expression by DNA-protein and protein-protein interactions in the promoter region of the zein multigene family. AB - A common cis-acting element in the promoter region of many genes expressed during endosperm development of cereal seeds, the prolamine-box or P-box, is only 20bp upstream of the alpha-class 22-kDa zein gene-specific cis element, the O2-box, which is recognized by the b-ZIP transcription factor, Opaque-2 (O2). The proximity of these two boxes has prompted a study of how two DNA-binding proteins of a different hierarchy might be involved in the activation and modulation of the 22-kDa zein-encoding genes. This was accomplished by utilizing a highly purified P-box-binding-factor-1 (PBF-1) and a bacterially expressed truncated form of the O2 protein. After adding the recombinant O2 to the purified fraction of PBF-1, binding studies were performed with a series of DNA probes combining the P- and O2-boxes from zein promoters. These studies have revealed an interesting inhibitory effect of PBF-1 over O2 function dependent on their ratio, consistent with its in-vivo properties and the developmental expression profiles of zein genes. We also could show that the P-box is specifically recognized by topoisomerase II and single-strand DNA-binding proteins, indicating a possible additional linkage between P-box and the scaffold-attachment-region (SAR). PMID- 9858762 TI - 'Interfacial affinity chromatography' of lipases: separation of different fractions by selective adsorption on supports activated with hydrophobic groups. AB - Lipases contained in commercial samples of lipase extracts from Rhizopus niveus (RNL) and Candida rugosa (CRL) have been selectively adsorbed on hydrophobic supports at very low ionic strength. Under these conditions, adsorption of other proteins (including some esterases) is almost negligible. More interestingly, these lipases could be separated in several active fractions as a function of a different rate or a different intensity of adsorption on supports activated with different hydrophobic groups (butyl-, phenyl- and octyl-agarose). Thus, although RNL seemed to be a homogeneous sample by SDS-PAGE, it could be separated, via sequential adsorption on the different supports, into three different fractions with very different thermal stability and substrate specificity. For example, one fraction hydrolyzed more rapidly ethyl acetate than ethyl butyrate, while another hydrolyzed the acetate ester 7-fold slower than the butyrate. Similar results were obtained with samples of CRL. Again, we could obtain three different fractions showing very different properties. For example, enantioselectivity for the hydrolysis of (R,S) 2-hydroxy-4-phenylbutanoic acid ethyl ester ranged from 1.2 to 12 for different CRL fractions. It seems that very slight structural differences may promote a quite different interfacial adsorption of lipases on hydrophobic supports as well as a quite different catalytic behavior. In this way, this new 'interfacial affinity chromatography' seems to be very suitable for an easy separation of such slightly different lipase forms. PMID- 9858763 TI - Structure and polymorphism of the Chironomus thummi gene encoding special lobe specific silk protein, ssp160. AB - cDNA encoding Chironomus thummi ssp160 was used to isolate a genomic clone that hybridized in situ to band A2b on polytene chromosome IV, the site of the ssp160 gene. DNA sequencing, primer extension and gene/cDNA nucleotide sequence alignment revealed the gene contains six exons and five introns; 70% of ssp160 is encoded in exon 3. Variations between cDNA and gene sequences led to the design of a polymerase chain reaction, restriction fragment length polymorphism assay that was subsequently used to demonstrate the existence of polymorphic alleles whose distribution varied between geographically separated populations of larvae. The polymorphism is associated with codon deletions in a six-amino-acid repeat containing an N-linked glycosylation motif. These deletions may have resulted from slipped-strand mispairing during DNA replication. PMID- 9858764 TI - 1H-NMR investigation of the influence of the heme orientation on functional properties of myoglobin. AB - There are two interconverting forms of myoglobin, which differ in the orientation of the heme by a 180 degrees rotation around the alpha,gamma-meso axis; the proteins possessing the same heme orientation, as found in the single crystal, and the reversed heme orientation are called the major and minor forms, respectively. Structures and functional properties of these two forms have been investigated by NMR. Heme peripheral side-chain and non-coordinated amino acid proton resonances of the minor form in its met-cyano form have been assigned and the comparison of the shift between the corresponding resonances of the two forms revealed that the heme electronic structure is not largely influenced by the heme rotation. On the other hand, the exchange rate of His E7 NepsilonH proton of the minor form is larger by a factor of 3-5 than that of the major one, indicating that the stability of the hydrogen bond between Fe-bound ligand and His E7 is slightly weaker in the minor form that in the major one. The rate of autoxidation of oxy-myoglobin and azide affinity of met-aquo myoglobin were also found to depend on the orientation of the heme. PMID- 9858766 TI - Analysis of the evolutionarily conserved repeat motifs in the genome of the highly endangered central Indian swamp deer Cervus duvauceli branderi. AB - We have analyzed the genome of central Indian swamp deer Cervus duvauceli branderi, an inhabitant of the Kanha National Park, a wildlife conservatory in Central India, with a view to provide a genetic basis for their extinction. Evolutionarily conserved repeat sequence motifs (GATA)3.75, TA(GATA)4, (GACA)3.75, (TGG)6 and a set of mouse beta-actin primers were used to uncover the sequence variation within and between related species by employing techniques of hybridization and AP-PCR amplification. The oligo probe carrying the GACA and TGG repeat motifs was found to be positive with Cervus genome, whereas (GATA)3.75, TA(GATA)4 and beta-actin probes did not cross-hybridize with the same. AP-PCR amplification with (GACA)3.75, unlike the (TGG)6 primer, generated distinct bands in the range of 0. 37-2.10kb amongst different genomes including Cervus. A comparative genome analysis of other species using the AP-PCR approach with (GACA)3.75 primer revealed the phylogenetic status of Cervus duvauceli branderi. From the analysis of a very limited number of Cervus DNA samples, we observed a high level of genetic homogeneity that may be a prime reason for the extinction of this species. This study has implications in the context of conservation of this endangered Cervus duvauceli branderi species. PMID- 9858765 TI - Characterization of the 3' untranslated region of mouse E2F1 mRNA. AB - The E2F family of transcription factors has been implicated in the regulation of the G1 to S phase transition of the mammalian cell cycle. We have focused on characterizing the cell cycle stage-specific expression of one family member, E2F1. Previous studies indicated that there are two mouse E2F1 (mE2F1) mRNA species whose abundance peaks in early S phase. However, it was unknown as to what constituted the structural difference between the two mE2F1 mRNAs and whether or not they encoded identical proteins. We have now cloned sequences corresponding to the 3' untranslated region (3'-UTR) of the mE2F1 gene. Northern blot analyses using different probes demonstrated that the two E2F1 mRNAs were distinguished by differences in the length of their 3' UTRs. We found that the longer (2.7-kb) mE2F1 mRNA contained two consensus RNA instability elements that the shorter (2.2-kb) mE2F1 mRNA lacked. However, a comparison of the stability of the 2.7-kb and the 2.2-kb mE2F1 mRNAs suggests that both mE2F1 mRNAs are fairly stable, having a half-life of 6-9h in both asynchronously growing cells and in the S phase of synchronized cells. Thus, we have determined that both mE2F1 mRNAs contain the identical coding region of the E2F1 protein and that enforced expression of mE2F1 mRNA should not be hampered by problems with RNA stability. PMID- 9858767 TI - pH dependence of the activation parameters for chymopapain unfolding: influence of ion pairs on the kinetic stability of proteins. AB - We studied the irreversible thermal denaturation of chymopapain, a papain-related cysteine proteinase. It was found that this process follows simple first-order kinetics under all conditions tested. Rate constants determined by monitoring ellipticity changes at 220 or 279 nm are essentially identical, indicating that denaturation involves global unfolding of the protein. Enthalpies (DeltaH(double dagger)) and entropies (DeltaS(double dagger)) of activation for unfolding were determined at various pH values from the temperature dependence of the rate constant. In the pH range 1.1-3.0, a large variation of both DeltaH(double dagger) and DeltaS(double dagger) was observed. For the few proteins studied so far (lysozyme, trypsin, barnase) it is known that activation parameters for unfolding vary little with pH. It is proposed that this contrasting behavior of chymopapain originates from the numerous ion pairs - especially those with low solvent accessibilities - present in its molecular structure. In contrast, fewer, more exposed ion pairs are present in the other proteins mentioned above. Our results were analyzed in terms of differences in the protonation behavior of carboxylic groups between the transition (TS) and native (N) states of the protein. For this purpose, a model of independently titrating sites was assumed, which explained reasonably well the pH dependence of activation parameters, as well as the protonation properties of native chymopapain. According to these calculations, pK values of carboxyls in TS are shifted 0.6-0.9 units upwards with respect to those in N. In addition, some groups in TS appear to be protonated with unusually large enthalpy changes. PMID- 9858768 TI - Characterization and mapping of four novel human expressed polymorphic trinucleotide microsatellites. AB - Recently, several important human genetic diseases have been shown to be due to pathological expansion of expressed trinucleotide microsatellites. Discovery of other such 'expansion diseases' will depend on characterization of more expressed loci containing trinucleotide repeats. We searched the expressed sequence tag database (dbEST) for repetitive trinucleotides and selected four loci for further studies (EST00586 EST05486, EST13299 and HHEA48B). For each locus, we have identified size polymorphism by PCR amplification and achieved chromosomal mapping using a somatic hybrid cell panel and the Stanford G3 radiation hybrid panel. Further searches of GenBank, dbEST and UniGene unraveled EST clusters for three of the loci, allowing construction of contigs and, in one instance, identification of a partial open reading frame. Three of the loci were linked to autosomal dominant human genetic diseases whose primary gene defect has not yet been established. Although it is a priori improbable that there is an etiological connection between the loci studied and the diseases, our results demonstrate that dbEST constitutes a useful starting point in the search for candidate loci for new expansion diseases. PMID- 9858769 TI - Inducible and constitutive transcription factors in the mammalian nervous system: control of gene expression by Jun, Fos and Krox, and CREB/ATF proteins. AB - This article reviews findings up to the end of 1997 about the inducible transcription factors (ITFs) c-Jun, JunB, JunD, c-Fos, FosB, Fra-1, Fra-2, Krox 20 (Egr-2) and Krox-24 (NGFI-A, Egr-1, Zif268); and the constitutive transcription factors (CTFs) CREB, CREM, ATF-2 and SRF as they pertain to gene expression in the mammalian nervous system. In the first part we consider basic facts about the expression and activity of these transcription factors: the organization of the encoding genes and their promoters, the second messenger cascades converging on their regulatory promoter sites, the control of their transcription, the binding to dimeric partners and to specific DNA sequences, their trans-activation potential, and their posttranslational modifications. In the second part we describe the expression and possible roles of these transcription factors in neural tissue: in the quiescent brain, during pre- and postnatal development, following sensory stimulation, nerve transection (axotomy), neurodegeneration and apoptosis, hypoxia-ischemia, generalized and limbic seizures, long-term potentiation and learning, drug dependence and withdrawal, and following stimulation by neurotransmitters, hormones and neurotrophins. We also describe their expression and possible roles in glial cells. Finally, we discuss the relevance of their expression for nervous system functioning under normal and patho-physiological conditions. PMID- 9858770 TI - Kinetic characterization of the inactivation of ammonia monooxygenase in Nitrosomonas europaea by alkyne, aniline and cyclopropane derivatives. AB - The kinetic mechanisms of seven inactivators of ammonia oxidation activity in cells of the nitrifying bacterium, Nitrosomonas europaea were investigated. The effects of the inactivators were specific for ammonia monooxygenase (AMO) which oxidizes ammonia to hydroxylamine. The aniline derivatives, 1,3-phenylenediamine and p-anisidine, were potent inactivators of AMO while other derivatives were ineffective as inactivators. Two cyclopropane derivatives, 1, 2 dimethylcyclopropane and cyclopropyl bromide, were inactivators while cyclopropane was not an inactivator. The mechanisms of three alkynes, 1-hexyne, 3 hexyne, and acetylene, were also examined. For all seven compounds, the inactivation of AMO was irreversible, time-dependent, first-order, and dependent on catalytic turnover. Saturation of the rate of inactivation was indicated for p anisidine (kinact=2.85 min-1; KI=1.0 mM) and cyclopropyl bromide (kinact=4.4 min 1; KI=97 microM), but not for any of the remaining five inactivators, including acetylene. Ammonia slowed the rate of inactivation for acetylene and cyclopropyl bromide, but enhanced the rate of inactivation for the remaining inactivators. All seven compounds appear to be mechanism-based inactivators of AMO. PMID- 9858771 TI - Amplification of erbB-4 oncogene occurs less frequently than that of erbB-2 in primary human breast cancer. AB - ErbB-4 protein is a recently discovered member of the ErbB family. The role of ErbB-4 protein in mammary-gland tissue has not been definitively established. To date, the expression of erbB-4 in breast tissue has been determined in only a few cases and, to the best of our knowledge, its amplification has not been examined. We therefore used the double differential polymerase chain reaction (ddPCR) for determination of the amplification profile of erbB-4 and erbB-2, another gene from the ErbB family, in human primary breast cancer specimens. We examined the amplification of the genes in 20 normal breasts and 176 invasive breast cancer samples. Amplification of erbB-2 was detected in 19% and erbB-4 in 13% of the samples studied. Co-amplification of the two oncogenes was found in only five out of 176 samples. Human breast cancer-derived cell lines in most cases overexpress both erbB-2 and erbB-4 (Beerli et al., 1995. Mol. Cell Biol. 15, 6496-6505; Han et al., 1995. Proc. Natl. Acad. Sci. USA 92, 9747-9751), but data on separate erbB-2 overexpression, without overexpression of erbB-4, were also reported (Wosikowski et al., 1997. Clin. Cancer Res. 3, 2405-2414). At the gene level, we found that co-amplification of the genes in the case of human breast cancer is rare. Moreover, an inverse association of the erbB-4 amplification with estrogen receptor activity and direct correlation with the tumor size were found. Due to these correlations, erbB-4 oncogene amplification can be assumed to be of prognostic or predictive value in the diagnosis of breast cancer. PMID- 9858772 TI - Single nucleotide polymorphism spectra in newborns and centenarians: identification of genes coding for rise of mortal disease. AB - Some single-nucleotide polymorphisms (SNPs) increase the risk of mortal disease. Identifying these SNPs and the genes in which they reside is an important area in human genomics. Such qualitative observations are important in themselves. However, an accurate assessment of the numerical distribution and age-dependent decline of SNPs in the population would permit calculation of the rises represented by each SNP. Such analyses have not been attempted because of a lack of an efficient and cost-effective method to detect multiple SNPs in a large number of individuals and a large number of genes. Here, we suggest the use of an analytical procedure that can scan for SNPs in 100-bp DNA sequences from as many as 10000 donors' blood cell samples, or 20000 alleles, simultaneously. Our suggestion is based on technology developed for studies of somatic mutations in human tissue DNA for point mutations at frequencies equal to or greater than 10( 6). In a simplified version of this technology, any SNP arising at frequencies at or above 5x10(-4) would be identified with useful precision. A gene would be represented by 10 or more sections of 100bp. This strategy includes splice-site mutations that represent a significant fraction of gene inactivating point mutations and would not be observed in strategies using cDNA. To illustrate the logic of the suggested approach, we use American mortality records to calculate the expected decrease in SNPs coding for premature mortality in newborns and centenarians. We consider several elementary cases: SNPs in one gene only, any of several genes, or all of several genes that create a risk of death by pancreatic cancer. The fraction of expressed polymorphisms affecting mortality should be simultaneously increased in probands and decreased in the aged relative to newborns. Silent polymorphisms in the same gene would remain unchanged in all three groups and serve as internal standards. A key point is that scanning a gene, in which loss of gene function creates the risk of mortality is expected to reveal not one, but multiple SNPs, which decline with age, as carriers die earlier in life than non-carriers. Several SNPs in a scanned gene would suggest that the decreasing SNP was genetically linked to a different polymorphism that creates the disease risk. PMID- 9858773 TI - Probing epitopes on human prorenin during its proteolytic and non-proteolytic activation. AB - The conformational changes of prorenin (PR) that are associated with its reversible non-proteolytic activation and irreversible proteolytic activation were monitored with immunoradiometric assays, using antibodies against epitopes belonging to the propeptide or the renin part of PR. Binding of PR to the renin inhibitor remikiren or protonation of PR resulted in the slowly progressive and simultaneous expression (t1/2 congruent with3.5-5.0 h at 4 degreesC) of epitopes of the N-terminal and C-terminal halves of the propeptide and an epitope that is manifest on renin but not on native non-activated PR. During reversible PR activation-inactivation, expression and disappearance of these epitopes coincided with the appearance and disappearance of enzyme activity. Cleavage of the propeptide from the renin part of PR by plasmin, as demonstrated by the failure of remikiren to unmask the N-terminal and C-terminal propeptide epitopes, was, with some time lag, followed by the simultaneous expression (t1/2 congruent with60 min at 4 degreesC) of the renin-specific epitope and enzymatic activity. Based on these findings we propose a model for the non-proteolytic activation of PR that involves the formation of an intermediary form of activated PR with the following properties: (1) the covalently bound propeptide has moved out of the active-site cleft, so that binding sites are exposed to active site ligands, (2) the propeptide is still not in the 'relaxed' conformation that is characteristic for fully, non-proteolytically, activated PR, and (3) the N-terminal part of the renin polypeptide chain has not yet attained the proper location that is required for enzymatic activity. PMID- 9858774 TI - Essential histidyl residues at the active site(s) of sucrose-phosphate synthase from Prosopis juliflora. AB - Chemical modification of sucrose-phosphate synthase (EC 2.4.1.14) from Prosopis juliflora by diethyl pyrocarbonate (DEP) and photo-oxidation in the presence of rose bengal (RB) which modify the histidyl residues of the protein resulted in the inactivation of the enzyme activity. This inactivation was dependent on the concentration of the modifying reagent and the time of incubation and followed pseudo-first order kinetics. For both the reagents, the inactivation was maximum at pH 7.5, which is consistent with the involvement and presence of histidine residues at the active site of the enzyme. Substrates, UDPG and F6P protected the enzyme against the inactivation by the modifying reagents suggesting that the histidine residues may be involved in the binding of these substrates and are essential for the catalytic activity. Specificity of DEP was indicated by an increase in absorbance at 240 nm along with concomitant inactivation of the enzyme and reactivation of the modified enzyme by hydroxylamine. These results strongly suggest the presence of histidine residue(s) at or near the active site of the enzyme. PMID- 9858775 TI - Lipoamide dehydrogenase from streptomyces seoulensis: biochemical and genetic properties. AB - Lipoamide dehydrogenase was purified around 22-fold relative to the crude extracts of Streptomyces seoulensis with an overall yield of 9. 5%. The enzyme was composed of two identical subunits with a molecular mass of 54 kDa and contained 1 mol of FAD per mol of subunit. The absorption spectra of the enzyme revealed the absorption maxima of flavoprotein at 272, 349, and 457 nm. Catalytically active two-electron reduced lipoamide dehydrogenase was produced by anaerobic reduction with one equivalent of NADH. Addition of excess amount of NADH led to the four-electron reduced lipoamide dehydrogenase. The reaction of the enzyme in the reduction reaction of lipoamide or lipoic acid could be explained by a ping-pong mechanism like many other lipoamide dehydrogenases reported earlier. The enzyme also catalysed the reduction of various quinone compounds with NADH as electron donor via a ping-pong mechanism. The enzyme can catalyse a single electron transfer in case of quinone-reducing process, evidenced by the production of 1, 4-naphthosemiquinone radical anion. The quinone reducing activity of the enzyme was dramatically inhibited by NAD+, indicating the involvement of four-electron reduced form. The structural gene for the enzyme was cloned using a DNA fragment PCR-amplified with the primers designed from N terminal and internal amino acid sequences. The deduced amino acid sequence shared striking similarity with those of lipoamide dehydrogenases from prokaryotes and eukaryotes. The gene was named lpd. All tested Streptomyces contained one homologue of the lpd gene, which is consistent with the fact that most organisms contain only one lipoamide dehydrogenase. PMID- 9858776 TI - Protein cross talking through osmotic work: the free energy of formation of the MgADP-myosin complexes at the muscle protein osmotic pressure. AB - A method is presented to determine the energy of formation of the myosin-ADP complexes at the muscle protein osmotic pressure. It is found that, at 18 kP, the putative protein osmotic pressure in skeletal muscle, the increase of MgADP from 0.05 to 2 mmolal, increases the free energy of myosin-ADP and of myosin-(ADP)2 by 0. 756 and by 9.85 kJ/mol, respectively, and decreases the free energy of myosin by 8.34 kJ erg/mol. It is pointed out that the local changes of water chemical potential, induced by the binding of MgADP to myosin, can be sensed by other structures of the contractile machinery, which per se may even be insensitive to MgADP. Cross talking between macromolecules can thus be achieved by changes of the water chemical potential. PMID- 9858777 TI - The homo-dimeric form of ADP-ribosyl cyclase in solution. AB - ADP-ribosyl cyclase is a multi-functional enzyme that catalyzes the formation of two Ca2+ signaling molecules, cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP). X-ray crystallography of three different crystal forms shows that it is a non-covalent dimer. Chemical cross-linking and dynamic light scattering were used in this study to determine if the cyclase is also a non-covalent dimer in solution. Treatment of the cyclase in dilute solution (0.05 mg/ml) with dimethylsuberimidate resulted in complete conversion to a species with molecular weight about twice that of the monomeric cyclase. Prolonged cross-linking of the cyclase at four times higher concentration produced also only the covalently linked dimers and no multimer formation was observed. The cross-linked dimer retained full enzymatic activity and readily catalyzed the formation of cADPR from NAD, NAADP from NADP, cyclic ADP-ribose phosphate from NADP, and cyclic GDP-ribose from nicotinamide guanine dinucleotide. Analysis of the autocorrelation functions obtained from dynamic light scattering measurements indicated the cyclase solution (2 mg/ml) was composed of a single molecular species and its diffusion coefficient was measured to be 7. 4x10-7 cm2/s. Computer modeling using the crystallographic dimensions of the non-covalent cyclase dimer, a donut shaped molecule with a central cavity and overall dimensions of 7x6x3 nm, gave a value for the diffusion coefficient essentially the same as that measured. These results indicate the cyclase is a non-covalent dimer in solution. PMID- 9858778 TI - High-potential states of blue and purple copper proteins. AB - Electrochemical measurements show that there are high-potential states of two copper proteins, Pseudomonas aeruginosa azurin and Thermus thermophilus CuA domain; these perturbed states are formed in guanidine hydrochloride (GuHCl) solution in which the proteins are still blue (azurin) and purple (CuA). In each case, the high-potential state forms reversibly. Absorption (azurin, CuA), visible circular dichroism (azurin, CuA), resonance-Raman (CuA), and EPR (CuA) spectra indicate that the structure of the oxidized copper site of each high potential form is very similar to that of the native protein. It is proposed that GuHCl perturbs one or more H-bonds in the blue or purple copper active site, thereby allowing Cu(I) to adopt a more favorable coordination structure than that in the rigid cavity of the native protein. PMID- 9858779 TI - Cloning of a thermostable ascorbate oxidase gene from Acremonium sp. HI-25 and modification of the azide sensitivity of the enzyme by site-directed mutagenesis. AB - A gene encoding a thermostable ascorbate oxidase (ASOM) was cloned from Acremonium sp. HI-25 and sequenced. The gene comprised 1709 bp and was interrupted by a single intron of 57 bp. ASOM consisted of 551 amino acids including a signal peptide with a molecular mass of 61200, and contained four histidine-rich regions with high sequence homology to the corresponding regions of other multicopper oxidases. The ASOM gene was expressed in Aspergillus nidulans under the Aspergillus oryzae Taka-amylase A gene promoter. The recombinant enzyme (An-ASOM) exhibited almost the same enzymatic properties as ASOM. The ASOM gene was mutated by site-directed mutagenesis with reference to the amino acid sequences of plant enzymes to generate enzymes with altered azide sensitivity. Site-directed mutagenesis at the trinuclear active copper site resulted in an increase in azide resistance; the Ala465Leu and Phe463Trp/Ala465Leu mutants exhibited approximately 10 and 20% increases in azide resistance, respectively. PMID- 9858780 TI - Subunit composition and oligomer stability of oat beta-glucosidase isozymes. AB - Oat beta-glucosidase (EC 3.2.1.21) has two isomeric forms, type I and type II, which are composed of 60 kDa peptides. To study the subunit composition and the stability of multimeric structure, the type I and II were purified from the primary leaves and coleoptiles of the etiolated oat seedlings where the isozymes are expressed organ-specifically. The monomers of the isozymes were isolated by urea-denatured gel electrophoresis followed by electroblotting. N-Terminal amino acid sequencing of the monomers indicated that the type I consisted of a peptide of ALESAKQVKPWQVPKRDWFP (As-Glu 1), and the type II having a peptide of ALESGKLKPWQIPKRDWFP (As-Glu 2) and As-Glu 1 in 1:1 ratio. The C-terminal amino acid of the As-Glu 1 was alanine and that of the As-Glu 2 was lysine. The As-Glu 2 was more negatively charged than the As-Glu 1. The type I isozyme is thus homomultimer of As-Glu 1 monomer and the type II heteromultimer of As-Glu 1 and As-Glu 2 monomers in 1:1 ratio. Partial denaturation of the multimers with urea and CaCl2 broke down the higher multimers to the lower multimers, which were in turn dissociated into homodimers and heterodimer. Denaturation study with urea and CaCl2 indicate that the higher multimers of the homooligomeric type I were more stable than those of the heterooligomeric type II and that hydrophobic interactions were important in the multimer formation. The homodimers were found to be more stable than the heterodimer. These results indicate that different combinations of the As-Glu 1 and As-Glu 2 monomers form the two isozymes of oat beta-glucosidase with different enzymatic properties and structural stability. PMID- 9858781 TI - In situ characterization of Helicobacter pylori arginase. AB - The properties of Helicobacter pylori arginase activity in metabolically competent cells and lysates were investigated with the aim of obtaining a better understanding of the nitrogen metabolism of the bacterium. One-dimensional 1H- and 13C-nuclear magnetic resonance spectroscopy, spectrophotometry, radio tracer analysis and protein purification techniques were employed to characterize in situ the first step in the utilization of l-arginine by the bacterium. Arginase activity was associated with the cell-envelope fraction obtained by centrifugation of lysates. A Km of 22+/-3 mM was determined for the enzyme activity, and differences of Vmax were observed between strains. Divalent cations stimulated arginase activity, and the most potent activators were Co2+>Ni2+>Mn2+. The activity was highly specific for l-arginine and did not catabolize analogs recognized by other arginases of prokaryote and eukaryote origin. The Ki of several inhibitors was measured and served also to characterize the enzyme activity. The presence of bicarbonate enhanced the hydrolysis of l-arginine in cell suspensions, but not in lysates or semi-purified enzyme preparations. Amino acid sequence analyses revealed important differences between the deduced structures of H. pylori arginase and those of other organisms. This finding was consistent with experimental data which showed that H. pylori arginase has unique properties. PMID- 9858782 TI - Exposure of the cryptic Arg-Gly-Asp sequence in thrombospondin-1 by protein disulfide isomerase. AB - Thrombospondin-1 is a matrix protein that inhibits proliferation, motility and sprouting of endothelial cells in vitro and angiogenesis in vivo. One mechanism by which thrombospondin-1 may influence endothelial cell biology is through interaction with the endothelial cell alphav beta3 integrin receptor. This interaction is mediated via a cryptic Arg-Gly-Asp sequence in the C-terminal Ca2+ binding region of thrombospondin-1. Exposure of the Arg-Gly-Asp sequence is controlled by disulfide interchange events in the Ca2+-binding loops and C globular domain. Limited reduction of thrombospondin-1 by dithiothreitol exposes the Arg-Gly-Asp sequence which can bind to the alphav beta3 integrin receptor and support endothelial cell spreading (X. Sun, K. Skorstengaard, D.F. Mosher, J. Cell Biol. 118 (1992) 693-701). Our aim was to identify possible physiological reductants that can mediate Arg-Gly-Asp exposure. We now report that protein disulfide isomerase, which is known to catalyze disulfide interchange in thrombospondin-1 and change its enzyme inhibitory properties and its binding to monoclonal antibodies, was secreted by bovine aortic endothelial cells and deposited on the cell surface. There was an average of approximately 2.2 fg of protein disulfide isomerase on the surface of a bovine aortic endothelial cell. Treatment of thrombospondin-1 with purified protein disulfide isomerase enhanced adhesion of endothelial cells to thrombospondin-1 in an Arg-Gly-Asp-dependent manner through the alphav beta3 integrin receptor and supported cell spreading. Both Ca2+-depleted and Ca2+-replete thrombospondin-1 were substrates for protein disulfide isomerase. These results suggest that endothelial cell derived protein disulfide isomerase may regulate Arg-Gly-Asp-dependent binding of thrombospondin 1. PMID- 9858783 TI - The function of Glu338 in the catalytic triad of the carbamoyl phosphate synthetase amidotransferase domain. AB - The synthesis of carbamoyl phosphate by the mammalian multifunctional protein, CAD, involves the concerted action of the 40 kDa amidotransferase domain (GLN), that hydrolyzes glutamine and the 120 kDa synthetase (CPS) domain that uses the ammonia, thus produced, ATP and bicarbonate to make carbamoyl phosphate. The separately cloned GLN domain has very low activity due to a reduction in kcat and an increase in Km but forms a hybrid complex with the isolated Escherichia coli CPS subunit. The hybrid has full glutamine-dependent catalytic activity and a functional interdomain linkage. The mammalian-E. coli hybrid was used to investigate the functional consequence of replacing His336 and Glu338, two residues postulated to participate in catalysis as part of a catalytic triad. The mutant mammalian GLN domains formed stable complexes with the E. coli CPS subunit, but the catalytic activity was severely impaired. While the His336Asn mutant does not form measurable amounts of the gamma-glutamyl thioester, the steady state concentration of the intermediate with the Glu338Gly mutant was comparable to the wild type hybrid because both the rate of formation and breakdown of the thioester are reduced. This result is consistent with the postulated role of Glu338 in maintaining His336 in the optimal orientation for catalysis and suggests a mechanism for the GLN CPS functional linkage. PMID- 9858784 TI - Limited proteolysis of a trypanosomal hypoxanthine phosphoribosyltransferase yields crystals that diffract X-rays to near atomic resolution. AB - Two crystal forms of the hypoxanthine phosphoribosyltransferase from Trypanosoma cruzi were grown and characterized. Proteolytic modification at the C-terminus of the recombinant enzyme yielded monoclinic crystals that diffract X-rays to higher resolution than the original, trigonal crystal form. Data from the monoclinic crystal form enabled determination of the crystal structure for the trypanosomal HPRT to 1.4 A resolution. PMID- 9858785 TI - Cloning of the mouse gene for D-dopachrome tautomerase. AB - D-Dopachrome tautomerase converts 2-carboxy-2,3-dihydroindole-5, 6-quinone (D dopachrome) into 5,6-dihydroxyindole. The amino acid sequence of this protein is 27% identical with that of macrophage migration inhibitory factor, which is known as a cytokine, pituitary hormone, and glucocorticoid-induced immunomodulator. In this study, we isolated and sequenced a 3490 bp-long genomic DNA of mouse D dopachrome tautomerase that consists of three exons and two introns. By two procedures, 5' rapid amplification of cDNA ends and cap site labeling, we determined the transcription initiation site, which is located 46 bp upstream of the translation initiation site. The possible polyadenylation sequence (AATAAA) is located 180 bp downstream of the termination codon. Computer-assisted analysis of the nucleotide sequence revealed a number of regulatory motifs, including multiple sites for Sp1, C/EBP, NF-Y, and USF. Although the precise pathophysiological functions of D-dopachrome tautomerase remain to be elucidated, the present results will contribute not only to elucidation of the mechanism of gene expression, but also to understanding of the molecular function of this protein. PMID- 9858786 TI - HspB3, the most deviating of the six known human small heat shock proteins. AB - From the alignment of 14 EST clones, the cDNA sequence of a novel human small heat shock protein (sHsp), called HspB3, could be deduced. The 3' part of the HspB3 cDNA is 99% identical to that of the previously reported HspL27 cDNA (W.Y. Lam, S.K. Wing Tsui, P.T. Law, S.C. Luk, K.P. Fung, C.Y. Lee, M.M. Waye, Isolation and characterization of a human heart cDNA encoding a new member of the small heat shock protein family-HSPL27, Biochim. Biophys. Acta 1314 (1996) 120 124). We argue that the HspB3 cDNA sequence is a corrected version of the HspL27 cDNA. The HspB3 cDNA is 742 bp long and contains an open reading frame specifying a polypeptide of 150 amino acid residues. Among the six known human sHsps it is evident that HspB3 is the most deviating one, having a unique N-terminal domain and essentially lacking a C-terminal extension. Northern blot analysis shows that in smooth muscle tissue the cDNA hybridizes with mRNA of about 0.9 kb. PMID- 9858787 TI - Hepatitis C virus and immunoglobulin gene rearrangements: an early step in lymphomagenesis? AB - A clonal expansion of peripheral blood mononuclear cells committed to IgM cryoprecipitating rheumatoid factor production has been demonstrated in hepatitis C virus (HCV)-associated mixed cryoglobulinemia (MC). To determine the role of HCV in B cell gene rearrangements we studied a series of 57 HCV-infected patients with and without MC. Clonal Ig gene rearrangements of both RNA and DNA were detected in 10 of the 13 patients with type II MC, 1 patient had gene rearrangement of the DNA only, and 2 had polyclonal patterns. 2 of the 17 patients with type III MC showed clonal rearrangement of both RNA and DNA, in 6 only the DNA was rearranged clonally and in 9 the patterns were completely normal. 14 of 27 patients with cryocrit <1% or without cryoglobulins had clonal DNA rearrangements without any in the RNA. These results suggest that clonal lesions in the DNA are related to HCV infection and that these changes antedate the appearance of mixed cryoglobulinemia. PMID- 9858789 TI - Determination of hematopoietic stem cells in peripheral blood by an automated hematology analyzer (SE-9000). AB - We evaluated the usefulness of an automated hematology analyzer (SE-9000) for the identification and counting of peripheral blood stem cells (PBSCs). The samples tested were from 14 patients with hematological malignancies. Peripheral blood samples were collected from the subjects before and after a course of chemotherapy. From the leukapheresis sample, CD34+ cells, assumed to be hematopoietic stem cells, were obtained with an immunomagnetic cell separator. The CD34+ cells obtained accumulated in the gate corresponding to low recurrent frequencies of the automated hematology analyzer. This gate shows results of the 'immature information' (IMI) channel. Software for detection of only the cells that accumulated in this gate was therefore developed. With this trial program, the regression coefficient between the percentage of leukocytes from the blood samples that were CD34+ and the percentage of such leukocytes that appeared on the IMI channel was 0.79. With this analyzer, the number of PBSC could be counted in about 80 s. The identification and counting of cells picked up by the IMI channel should be clinically useful for the monitoring of changes in PBSC after chemotherapy for mobilization. PMID- 9858788 TI - Rabbit IgG antibodies against cord red blood cell membranes bind to complement receptor 1 (CD35). AB - We have previously shown that a subpopulation of cord/fetal red blood cells (RBC) binds rabbit IgG antibodies raised against cord RBC and absorbed on adult RBC (F IgG), while control IgG, raised against and absorbed on adult RBC (A-IgG), fails to do so. In the present study, F-IgG maintained its binding to cord RBC surface antigens following absorption on spectrin but not after absorption on skeleton stripped RBC membranes. Spectrin-absorbed F-IgG- but not A-IgG-affinity-purified material from cord RBC contained polypeptides with apparent MW of complement receptor 1 (CR1) allotypes. Moreover, on immunoblotting these polypeptides reacted with 125I-F-IgG as well as with 125I-anti-CR1 mAb, and binding of 125I anti-CR1 mAb was inhibited by unlabelled F-IgG. In addition, cord RBC incubated with F-IgG prior to reaction with anti-CR1 showed decreased fluorescence intensity on flow cytometry. Taken together the results suggest that F-IgG binds to CR1 which shows increased expression/accessibility on a subpopulation of cord/fetal RBC. PMID- 9858790 TI - Changes in serum thrombopoietin levels after splenectomy. AB - To clarify the role of thrombopoietin (c-Mpl ligand, TPO) in 'hypersplenic' thrombocytopenia, we used an enzyme-linked immunosorbent assay to examine changes in serum TPO levels accompanied with splenectomy in 6 patients with liver cirrhosis, 4 patients with gastric cancer, and 2 patients with lymphoid malignancies. We also measured serum levels of other thrombopoietic cytokines such as interleukin-6 (IL-6) and erythropoietin. Platelet counts reached a maximum at day 14 after splenectomy in all subjects. In patients with liver cirrhosis, a lower elevation of platelet counts was observed compared with that in patients with gastric cancer. Serum TPO levels gradually elevated after splenectomy and reached a maximum 3.5 days after splenectomy in noncirrhotic patients, whereas peak serum TPO levels were delayed until day 7 in the cirrhosis group. IL-6 and erythropoietin showed similar kinetics between cirrhotic and noncirrhotic patients. These findings suggest that transient thrombocytosis after splenectomy may be associated with an alteration in the site of TPO catabolism by platelets from spleen to the blood and that deterioration of TPO production may play a role in thrombocytopenia in liver cirrhosis. PMID- 9858791 TI - Some aspects of morphological and dysplastic changes of the bone marrow in malignant lymphoma. AB - Cytological and histological study of the bone marrow of 33 patients with malignant lymphoma was performed to evaluate (1) the occurrence of secondary myelodysplastic changes, (2) the cell lines involved and (3) whether secondary myelodysplastic changes were similar or different for non-Hodgkin's lymphoma (NHL) and Hodgkin's disease (HD). Different changes in erythro-, granulo- and megakaryopoiesis were revealed. Granulo- and erythropoiesis were disturbed more frequently. Morphological changes of the erythroid series, megaloblastoid maturation of the nucleus and multinuclearity, were seen more often in NHL patients. Cells of the granulocytic series demonstrated different changes: hypogranulation and hypersegmentation in NHL, hypergranulation and Dohle's bodies in HD. No remarkable dysplastic changes of megakaryopoiesis were demonstrated. PMID- 9858792 TI - Influence of alpha-thalassemia on cholelithiasis in SS patients with elevated Hb F. AB - Chronic hemolysis, with consequent hyperbilirubinemia, predisposes SS patients to pigment gallstones. The other factors which influence the development of stones in these patients have not been identified. We have carried out a combined prospective and retrospective study of SS patients in Kuwait and specifically investigated the influence of coexistent alpha-thal trait on the prevalence of gallstones. A total of 45 patients (30 males, 15 females) with ages ranging from 1 to 16 years (mean 7.2 +/- 3.1) were studied. Most were either homozygotes for the Saudi Arabia/India haplotype (86.7%) or compound heterozygotes for this and the Benin haplotype (11.1%). They were screened for gallstones with ultrasonography. alpha-Globin genotypes were determined using a combination of PCR and allele-specific oligonucleotide hybridization techniques to identify the common alpha-thalassemia alleles in this population. Gallstones were detected in 7 (15.6%) patients (4 males, 3 females), whose mean age (10.5 +/- 5.5 years) was significantly higher than that (6.8 +/- 3.2 years) of those without stones (p < 0.01). The mean total Hb of the former (8.4 +/- 0.8 g/dl) was also significantly (p < 0.05) lower than in the latter (9.5 +/- 1.3 g/dl), while the difference in mean Hb F levels was not significant. None of the 4 alpha-thal homozygotes had gallstones while 2 of 13 heterozygotes and 5 of the 23 patients without coexistent alpha-thal had. The differences in these proportions are statistically significant (chi2 = 20.4, p < 0. 001). It therefore appears that coexistent alpha thal decreases the chance of developing gallstones in Arab SS patients. This may be related to less hemolysis in such patients as shown by their higher mean Hb level. PMID- 9858793 TI - Immunoblastic lymphadenopathy-like T cell lymphoma evolving into a massive plasma cell proliferation with biclonal paraproteinemia. AB - We present a case of immunoblastic lymphadenopathy-like T cell lymphoma (IBL-T) who subsequently developed a massive proliferation of plasma cells. At diagnosis of IBL-T, the patient had polyclonal hypergammaglobulinemia and subsequently, while on chemotherapy, developed paraproteinemia with biclonal peaks and the IBL T lesion was replaced with a massive proliferation of CD38-positive plasma cells. The evolution was not likely to be attributed to a new neoplastic proliferation of B cells. It appeared that two B cell clones possibly had a growth advantage among the polyclonal B cells due to a depletion of suppressor T cells or to a disturbance in the immune system. PMID- 9858794 TI - Pseudoreticulocytosis in a case of myelodysplastic syndrome. AB - A patient with normocytic anemia and marked and persistent reticulocytosis is presented. Causes responsible for blood loss and hemolytic diseases were excluded and, after bone marrow examination, myelodysplastic syndrome (refractory anemia) was diagnosed. In vitro reticulocyte survival studies suggested that reticulocytosis was a consequence of delayed maturation of the reticulocytes. Pseudoreticulocytosis may be an unusual presentation of myelodysplastic syndromes, because only 4 patients with such a finding have previously been reported. PMID- 9858795 TI - Cyclosporine and entrapment neuropathy. Report of two cases. PMID- 9858796 TI - Hemophagocytic ability of rhabdomyosarcoma cells. PMID- 9858797 TI - L-Carnitine treatment in beta thalassemia major. PMID- 9858798 TI - Angioimmunoblastic lymphadenopathy: treatment in three cases. PMID- 9858799 TI - Thromboembolism in beta-thalassemia major. PMID- 9858800 TI - Different brain morphologies from different genotypes in a single teleost species, the medaka (Oryzias latipes) AB - In a teleost fish, the medaka (Oryzias latipes), many inbred strains have been established from various origins including wild populations. Brains from five genetically different strains, which had been bred and raised under the same conditions, were examined to determine whether there is intraspecific genetic variation. A total of 25 brains from the wild-type strains (HNI-II, HB11A and HB32C) and from the body-colour mutant strains (Hi3 and HO5) were fixed, and the external features of the brains were examined under a stereomicroscope. The differences between the HNI-II brains and the Hi3 brains were the most remarkable in the external features. In order to carry out a volumetric analysis, the brains of all strains were cut into complete serial cryostat sections. Total brain volumes and relative volumes (in % of total brain volume) of the olfactory bulb, telencephalon, optic tectum, and cerebellum were calculated in each brain using a semi-automatic image analyzer. Statistical analysis showed that significant differences in the total brain volumes and the relative volumes of these subdivisions exist not only between wild-type and mutant strains but also among wild-type strains. Thus, our results demonstrate that the strains with different genotypes possess large variation in brain morphology. This is the first report to demonstrate that there exists intraspecific genetic variation in the gross brain morphology of a wild-type vertebrate. PMID- 9858801 TI - Calbindin immunoreactivity in the auricular lobe and interauricular granular band of the cerebellum in bullfrogs. AB - Calcium binding protein (CaBP) immunoreactivity in the cerebellum of bullfrogs was examined, concentrating on cells associated with the auricular lobe. While anti-calretinin and anti-parvalbumin also immunoreacted with the same cell populations, anti-calbindin exhibited the most robust and typical pattern of immunostaining. Calbindin immunoreactivity was observed in various populations of cells in the auricular lobe and interauricular granular band of the cerebellum, in the cerebellar peduncle, and in a bundle of interauricular commissural fibers which course through the dorsal, marginal, part of the molecular layer. Cells in the granular layer of the ventral part (i.e., corpus cerebelli) of the cerebellar plate were not CaBP-immunoreactive, nor were any fibers in the molecular layer of this cerebellar region. We believe that axons of CaBP-immunoreactive granule-like cells of the auricular lobes contribute to the formation of the interauricular fiber bundle, which corresponds to the lateral commissure of urodele amphibians. The pattern of calbindin immunoreactivity in the auricular lobes and marginal part of the cerebellar plate provides additional evidence that this cerebellar compartment, which is already present in tadpoles, has a distinct origin, biochemical characterization and connectivity and is separate from the compartment that forms the corpus cerebelli of frogs during metamorphosis. PMID- 9858802 TI - Rattlesnake hunting behavior: correlations between plasticity of predatory performance and neuroanatomy. AB - Rattlesnakes may shift between visual (eyes) and infrared (facial pits) stimuli without significant loss of predatory performance during an envenomating strike. The relative equivalency of these proximate stimuli is correlated with the organization of the associated neural pathways in the central nervous system. Visual and infrared information, although gathered by different sensory organs, converges within the optic tectum in an orderly spatiotopical representation where bimodal neurons respond to both stimuli. In turn, the tectum sends efferent pathways directly to premotor areas (brainstem) and indirectly to motor areas (spinal cord) where axial muscles involved in the strike might be activated. On the other hand, rattlesnakes do not maintain a high level of equivalent predatory performance when switching between chemosensory stimuli i.e., olfactory, and vomeronasal information. Deprived of vomeronasal input, strikes drop by about half, and poststrike trailing is lost entirely. Surprisingly, compensation by switching to information delivered via an intact olfactory input does not occur, despite the convergence of chemosensory information within the central nervous system. Finally, the launch of a targeted, envenomating strike involves both these modalities: radiation reception (visual, infrared) and chemoreception (olfactory, vomeronasal). However, in the absence of chemosensory information, the radiation modalities do not completely compensate, nor does the animal maintain a high level of predatory performance. Similarly, in the absence of radiation information, the chemosensory modalities do not completely compensate, nor does the animal maintain a high level of predatory performance. The absence of compensation in this multimodal system is also correlated with an absence of convergence of radiation and chemical information, at least at the level of first and second-order neurons, in the central nervous system. PMID- 9858803 TI - Diurnal and nocturnal visual capabilities in shorebirds as a function of their feeding strategies. AB - Some shorebird species forage with the same feeding strategy at night and during daytime, e.g. visual pecking in the Wilson's Plover (Charadrius wilsonia) or tactile probing in the Short-billed Dowitcher (Limnodromus griseus). The American Woodcock (Scolopax minor) uses tactile probing, by day and by night, but sometimes pecks for insects during daytime. The Black-winged Stilt (Himantopus himantopus) is a visual pecker, both by day and by night, and sometimes forages tactilely on windy (agitated water surface) moonless nights. Territorial Willets (Catoptrophorus semipalmatus) are visual peckers during daylight and on moonlight conditions but switch to tactile feeding under lower light conditions. It could be postulated that some shorebird species would switch from visual feeding during daytime to tactile foraging at night because they have poor night vision compared to species that are always sight foragers irrespective of the time of the day. This issue was examined by comparing retinal structure and function in the above species. Electroretinograms (ERGs) were obtained at different light intensities from anesthetized birds, and the retinae were processed for histological observations. Based on ERGs, retinal sensitivity, and rod:cone ratios, both plovers and stilts are well adapted for nocturnal vision. Although they have low rod density compared to that of stilts and plovers, Willets and woodcocks have a scotopic retinal sensitivity similar to that of stilts and plovers but rank midway between plovers and dowitchers for the b-wave amplitude. Dowitchers have the lowest scotopic b-wave amplitude and retinal sensitivity and appear the least well adapted for night vision. Based on photopic ERGs and cone densities, although stilts, Willets and dowitchers appear as well adapted for daytime vision, plovers occupy the last rank of all species examined. Compared to the nighttime tactile feeders and those that switch from daytime visual pecking to tactile feeding at night, nighttime sight feeders have a superior rod function and, consequently, potentially superior nocturnal visual capabilities. PMID- 9858804 TI - Comparative anatomy of the claustrum in selected species: A morphometric analysis. AB - The morphology of the claustrum was studied by stereological methods in representatives of five mammalian orders (Insectivora, Rodentia, Lagomorpha, Carnivora and Primates). In each species under study, a dorsal and a ventral part of the nucleus can be distinguished. Based on differences in shape and separation from surrounding structures, five morphological types of the claustrum occur. The claustrum of Insectivora and some rodents represents the least complicated morphological type. The nucleus is very poorly separated from the surrounding structures. The human claustrum is morphologically the most complicated, although the two above-mentioned principal divisions are apparent. The ventrally situated paraamygdalar part of the human claustrum may correspond to the endopiriform nucleus or ventral part of the claustrum of other mammals, because of its morphological characteristics and connections with the limbic system. In guinea pigs, traditionally classified as members of the Rodentia, a characteristic morphological type of the claustrum is present. This observation may support arguments questioning the current position of this species in mammalian classification. Based on stereological studies, the increase of the claustral volume that occurs with increase of the hemispheric volume is significantly smaller than the increase of the isocortical volume and larger than the increase of the allocortical volume. The increase of the volume of the dorsal and ventral parts of the claustrum does not differ significantly in the species under study. Neurons of the claustrum represent differentiated morphology. The numerical density of neurons in the dorsal part of the claustrum is significantly higher than in the ventral one. Differences in the morphology and cellular structure of the two parts of the claustrum may suggest differences in function of the two parts of the nucleus, most probably concerned with transfer of information among various cortical regions. Changes in the claustrum, a cortico-related structure, that occur with increased brain volume, may suggest that its development is less dynamic than that of the isocortex. PMID- 9858805 TI - Assignment1 of inosine '-monophosphate dehydrogenase type 2 (IMPDH2) to human chromosome band 3p21.2 by in situ hybridization. PMID- 9858806 TI - Identification and characterization of STK12/Aik2: a human gene related to aurora of Drosophila and yeast IPL1. AB - Mutations in aurora of Drosophila and related Saccharomyces cerevisiae IPL1 protein kinases are known to cause abnormal chromosome segregation. We earlier isolated a cDNA encoding a novel human protein kinase Aik which shares high amino acid identity with the Aurora/Ipl1 protein kinase family. In the present study, a second human cDNA highly homologous to aurora/IPL1 (Aik2) was identified and the nucleotide sequence was determined (gene symbol STK12). The C-terminal kinase domain of the STK12 encoded protein shares high amino acid sequence identity with those of mouse STK-1 (90%), rat AIM-1 (90%), human Aik (69%), mouse IAK1/Ayk1 (69%), Xenopus pEg2 (68%), Drosophila Aurora (62%), and yeast Ipl1 (45%), whereas the N-terminal domain of the STK12 protein shares little homology with those of Aurora/Ipl1 family members except for AIM-1 and STK-1. Northern blotting analyses revealed that STK12 expression was high in thymus, while low level expression was detected in small intestine, testis, colon, spleen, and brain. The STK12 protein content in HeLa cells is low in S phase, but it accumulates during M phase. STK12 was mapped to human chromosome 17p13.1 by fluorescence in situ hybridization. The chromosome location of STK12 was further defined using a radiation hybrid panel (Stanford G3), that showed a linkage with marker WI-7901 (LOD Score 7.83) located between D17S938 and D17S786. PMID- 9858807 TI - Assignment of Ptprn2, the gene encoding receptor-type protein tyrosine phosphatase IA-2beta, a major autoantigen in insulin-dependent diabetes mellitus, to mouse chromosome region 12F. AB - The receptor-type protein tyrosine phosphatase IA-2beta gene (mouse gene symbol Ptprn2) encodes a major autoantigen in insulin-dependent diabetes mellitus. We physically mapped Ptprn2 by fluorescence in situ hybridization to band F of mouse chromosome 12, a region that lacks diabetes susceptibility loci. The mapping confirms the proposed synteny of mouse 12F with band q36 of human chromosome 7. PMID- 9858808 TI - Genomic analysis of DUSP6, a dual specificity MAP kinase phosphatase, in pancreatic cancer. AB - DUSP6 (alias PYST1), one of the dual-specificity tyrosine phosphatases, is localized on 12q21, one of the regions of frequent allelic loss in pancreatic cancer. This gene is composed of three exons, and two forms of alternatively spliced transcripts are ubiquitously expressed. Although no mutations were observed in 26 pancreatic cancer cell lines, reduced expressions of the full length transcripts were observed in some cell lines, which may suggest some role for DUSP6 in pancreatic carcinogenesis. PMID- 9858809 TI - An optimized, fully automated system for fast and accurate identification of chromosomal rearrangements by multiplex-FISH (M-FISH). AB - Multiplex-FISH (M-FISH) is a recently developed technique by which each of the two dozen human chromosomes-the 22 autosomes and the X and Y sex chromosomes-can be stained or "painted" with uniquely distinctive colors. Using a combinatorial labeling technique and a specially designed filter set, each DNA probe can be identified by its unique spectral signature. Here we present several significant optimizations of the M-FISH technology. First, a new strategy for labeling the probes is described which allows for easy and fast production of the complex M FISH probe mix. Second, a newly developed, completely motorized microscope equipped with an eight-position filter wheel and a new generation of filter sets is presented that allows fully automatic imaging of a complete metaphase spread within seconds. Third, to determine the characteristic spectral signatures for all different combinations of fluorochromes, we developed a novel multichannel image analysis method. The spectral analysis is solely guided by the image information itself and does not require any user interaction. A complete analysis of a metaphase spread can be accomplished in less than 3 min. Sophisticated built in quality controls were developed, and the value of visual inspection of M-FISH images as a simple means of controlling the computer-generated chromosome classification are illustrated. In addition, we discuss advantages of adding new fluorochromes to the traditionally used five fluorochromes. PMID- 9858810 TI - A versatile image analysis approach for simultaneous chromosome identification and localization of FISH probes. AB - Modern cytogenetic techniques, such as comparative genomic hybridization (CGH) and the multi-color fluorescence in situ hybridization (FISH) techniques of multiplex fluorescence in situ hybridization (M-FISH) and spectral karyotyping (SKY), require a coordinated banding analysis to maximize their usefulness. All of the methods currently used, including Giemsa (G-) banding, Alu banding, and 4',6-diamidino-2-phenyl-indole (DAPI) banding, have serious drawbacks. A simple and effective method to band chromosomes concurrently with FISH is needed. To address this problem, we stained chromosomes with DAPI and chromomycin A3, and then used an image analysis program to generate banding by dividing the image taken with a DAPI excitation filter by the image taken with a chromomycin A3 excitation filter. The result was a metaphase spread in which the chromosomes possessed a banding pattern characteristic of R-banding. The image analysis program was then used to generate linescans of pixel intensity versus relative position along the length of chromosomes that were banded using this technique, which we have called D/C R-banding. Each chromosome in a genome was represented by a characteristic scan profile, which was unaffected by FISH signals. Reference linescans were prepared by karyotyping D/C R-banded chromosomes for a given species, and then drawing lines along the length of the known chromosomes. The linescans were combined into a spreadsheet database, which was linked by dynamic data exchange to the image analysis program and normalized for length and intensity. The linescan of an unknown chromosome was then transferred to the spreadsheet, where it was normalized for length and intensity and overlaid on the linescans of each chromosome in the genome. Unknown chromosomes were identified by comparison of their graphs with graphs in the standardized reference genome. We have used this approach to create reference linescan karyotypes of several species, and to identify chromosomes on which FISH was performed. PMID- 9858811 TI - Mouse centromere protein F (Cenpf) gene maps to the distal region of chromosome 1 by interspecific backcross analysis. PMID- 9858812 TI - Construction of a whole-genome radiation hybrid panel for high-resolution gene mapping in pigs. AB - We have developed a panel of 152 whole-genome radiation hybrids by fusing irradiated diploid pig lymphocytes or fibroblasts with recipient hamster permanent cells. The number and size of the porcine chromosome fragments retained in each hybrid clone were checked by fluorescence in situ hybridization with a SINE probe or by primed in situ labeling (PRINS) with SINE-specific primers. A strategy based on the interspersed repetitive sequence polymerase chain reaction (IRS-PCR) was developed for selected clones to determine if the large fragments painted by the SINE probe corresponded to one pig chromosome or to different fragments of several chromosomes. This strategy was buttressed by a double PRINS approach using primers specific for alpha-satellite sequences of two different groups of swine chromosomes. Genome retention frequency was estimated for each clone by PCR with 32 markers localized on different porcine chromosomes. Of the 152 hybrids produced, 126 were selected on the basis of cytogenetic content and chromosome retention frequency to construct a radiation hybrid map of swine chromosome 8. Our initial results for this chromosome indicate that the resolution of the radiation hybrid map is 18 times higher than that obtained by linkage analysis. PMID- 9858813 TI - Molecular cytogenetic mapping of 24 CEPH YACs and 24 gene-specific large insert probes to chromosome 17. AB - Defining boundaries of chromosomal rearrangements at the molecular level would benefit from landmarks that link the cytogenetic map to physical, genetic, and transcript maps, as well as from large-insert FISH probes for such loci to detect numerical and structural rearrangements in metaphase or interphase cells. Here, we determined the locations of 24 genetically mapped CEPH-Mega YACs along the FLpter scale (fractional length from p-telomere) by quantitative fluorescence in situ hybridization analysis. This generated a set of cytogenetically mapped probes for chromosome 17 with an average spacing of about 5 cM. We then developed large-insert YAC, BAC, PAC, or P1 clones to the following 24 known genes, and determined refined map locations along the same FLpter scale: pter-TP53-TOP3-cen TNFAIP1-ERBB2-TOP2A- BRCA1-TCF11-NME1-HLF-ZNF147/CL N80-BCL5/MPO/SFRS1-TBX2 PECAM1-DDX5/ PRKCA-ICAM2-GH1/PRKAR1A-GRB2-CDK3 /FKHL13-qter. Taken together, these 48 cytogenetically mapped large-insert probes provide tools for the molecular analysis of chromosome 17 rearrangements, such as mapping amplification, deletion, and translocation breakpoints in this chromosome, in cancer and other diseases. PMID- 9858814 TI - Human chromosome 9 pericentric homologies: implications for chromosome 9 heteromorphisms. AB - Pericentromeric polymorphisms of chromosome 9 include variations in the size of heterochromatin, pericentric inversions, and, more rarely, additional C-band negative, G-band-positive material in either the proximal short arm or long arm or within the heterochromatin. It has been postulated that rearrangements involving the different classes of satellite DNA present in this relatively unstable region of the human genome constitute a mechanism for the origin of these variants. We report the identification, by molecular cytogenetic investigations, of homologous stretches of euchromatin shared by the proximal short and long arms of chromosome 9 that suggest that exchanges involving these regions may be an additional mechanism for the origin of chromosome 9 polymorphisms. PMID- 9858815 TI - Evidence of a differential organization of chromatin containing terminal or interstitial (TTAGGG)n repeats by in situ digestion with nucleases. AB - (TTAGGG)n sequence repeats in human telomeres and in Chinese hamster interstitial centromeric areas were digested in situ with exonuclease III (ExoIII) and exonuclease Bal 31. Incubation with AluI was performed beforehand to increase DNA breaks near telomere sequence areas. DNA removal at these specific regions was quantified by digital image analysis of the fluorescence in situ hybridization signal produced by a telomeric probe. Exonuclease III was 2.6 times more active in interstitial than in terminal telomeric sequence areas. Exonuclease Bal 31 was 2.3 times more effective in terminal than in interstitial telomeric sequence regions. These results support the hypothesis that chromatin is differentially organized in both telomeric sequence areas, despite their similar DNA composition. PMID- 9858816 TI - Refined chromosomal localization of the putative tumor suppressor gene TP73. AB - We report the refined chromosomal localization of the putative tumor suppressor gene TP73 (alias p73) within the genomic region between the anonymous loci D1Z2 and D1S47. The region measures less than 6 Mb and covers a genetic distance of 16 cM. The present mapping considerably restricts the previous cytogenetic localization of TP73. PMID- 9858817 TI - Assignment of gamma-glutamyltransferase (GGT) to porcine chromosome band 14q2.1 by in situ hybridization. PMID- 9858818 TI - Sperm segregation analysis of a complex chromosome rearrangement, 2;22;11, by whole chromosome painting. AB - Using the human sperm-hamster oocyte fusion technique and whole chromosome painting, we studied sperm chromosome segregation in a male heterozygous for a complex chromosome rearrangement, 46,XY,-2, +der(2)t(2;11)(q13; q23), 11,+der(11)t(11;22)(q23;q11.2),-22, +der(22)t(2;22)(q13; q11.2). A total of 208 sperm complements were analyzed. The frequency of sperm carrying a normal or a balanced complement was 13.5% (9.62% and 3.85%, respectively). The frequency of unbalanced sperm was 86.5% (64.9% from 3:3 segregation, including 30 different types; 20.7% from 4:2 segregation, including 21 different types; and 0.96% from 5:1 segregation, including 2 different types). The sex ratio, determined in 134 sperm complements, did not differ from the expected 1:1 ratio. The results obtained in this study are compatible with the formation, during the synaptic process, of a complex hexavalent figure involving chromosomes 2, 11, and 22. The behavior and segregation of this complex figure would explain the high frequency (86.5%) of unbalanced complements observed in this carrier. PMID- 9858819 TI - ZOO-FISH and R-banding reveal extensive conservation of human chromosome regions in euchromatic regions of river buffalo chromosomes. AB - Commercially available human chromosome (HSA) painting probes were hybridized on river buffalo (Bubalus bubalis, 2n = 50) chromosomes by using FISH and R-banding techniques. Clear hybridization FITC-signals revealed extensive conservation of human chromosome regions in this species and demonstrated that human chromosome probes primarily paint euchromatic regions (R-bands). The present results are discussed in the light of previous gene mapping data obtained in river buffalo and ZOO-FISH data in cattle, and in relation to the standard bovine chromosome nomenclatures. In particular, HSA 8, HSA 10, HSA 11, and HSA 16+7 paint, respectively, BBU 1p, BBU 4p, BBU 5p, and BBU 24, which are homoeologous, respectively, to cattle chromosomes 25, 28, 29 and 27. Thus, these river buffalo chromosome arms can serve as markers to resolve discrepancies in the nomenclature of cattle and related species. PMID- 9858820 TI - Optimized mitogen stimulation induces proliferation of neoplastic B cells in chronic lymphocytic leukemia: significance for cytogenetic analysis.The Tampere Chronic Lympocytic Leukemia group. AB - We tested the effects of interleukin-2 (IL-2), human recombinant tumor necrosis factor alpha (TNF-alpha), Staphylococcus aureus Cowan I (SAC), TPA, and their combinations, using a standard thymidine incorporation assay, in order to identify an optimal mitogen combination (OMC) for 24 consecutive patients with B cell chronic lymphocytic leukemia (B-CLL). The combination that induced the highest thymidine incorporation was chosen as the OMC for each patient. Among 14 mitogen combinations tested, there were six different OMCs, of which the most frequent was TNF-alpha + IL-2. It was the OMC in 9 of 24 cases. The other OMCs were TNF-alpha + TPA1 (5/24), SAC + IL-2 (5/24), TPA1 + IL-2 (3/24), TPA10 + IL-2 (1/24), and TNF-alpha + TPA10 + IL-1 (1/24). The mitogenic power of the selected OMC in each case was then evaluated both by the combination of immunophenotyping and molecular cytogenetic techniques known as MAC (Morphology, Antibody, Chromosomes) and standard chromosome analysis. After OMC stimulation, the levels of DNA synthesis and B-cell proliferation (mitotic index) were, on average, 10 fold higher than those observed after standard TPA stimulation (P < 0. 0001). The proportion of mitotic B cells exceeded the proportion of mitotic T cells in 70.1% of the cases after OMC stimulation. After TPA stimulation, 7.7% +/- 2.5% of all mitoses were B-cell mitoses, whereas after OMC stimulation this proportion rose to 57.9% +/- 5.3%. The frequency of clonal chromosomal aberrations increased from 46% after TPA stimulation to 79% after OMC stimulation. The clonal aberrations del(6q), del(11q), and/or del(13q) were observed in 26%, 32%, and 42% of the patients with the respective clonal chromosomal aberrations, whereas the corresponding frequencies after TPA stimulation were only 4%, 21%, and 17%. When the lineage involvement of cells with clonal chromosomal aberrations from three patients was analyzed, the aberrations were found to be restricted to B cells only, and in one patient to a minor subset of B cells. The results demonstrate that an individually chosen OMC induces a high rate of proliferation in neoplastic B cells. We found deletions in 6q, 11q, and 13q at higher frequencies than reported previously, most probably as a result of an improved mitogenic response. The identification of an optimal mitogen stimulation for each patient, prior to chromosome analysis, can well be expected to reduce the rate of false normal results in the future. This is essential for accurate evaluation of the prognostic significance of chromosomal aberrations in B-CLL. PMID- 9858821 TI - Assignment of Sptlc2 to murine chromosome band 12, region E by in situ hybridization. PMID- 9858822 TI - Assignment of human PLD1 to human chromosome band 3q26 by fluorescence in situ hybridization. PMID- 9858823 TI - Assignment of human PLD2 to chromosome band 17p13.1 by fluorescence in situ hybridization. PMID- 9858824 TI - Assignment of the cellular retinol-binding protein 1 gene (RBP1) and of the coatomer beta subunit gene (COPB2) to human chromosome band 3q23 by in situ hybridization. PMID- 9858825 TI - Assignment of SHOX2 (alias OG12X and SHOT) to human chromosome bands 3q25-->q26.1 by in situ hybridization. PMID- 9858826 TI - Characteristics of chromosomes in polarized normal human bronchial cells provide a blueprint for nuclear organization. AB - In normal human terminally differentiated and polarized bronchial cells, the fluorescent "painting" technique (FISH) for all chromosomes (except Y) documented that each homologue of each chromosome occupies a distinct, separate domain within the nucleus. The homologues are distributed along the nuclear membrane. In most cells and chromosomes studied, the two homologues were not identical: one was usually more "compact" than the other which was more "open," displaying fiber shaped extensions. The differences between the territories of homologues 1 and 7 were shown to be statistically valid (P < 0.0001 by Wilcoxon sign rank test), as has been previously documented for the two X chromosomes (Eils et al., 1996). In some parallel arrays of bronchial cells, the position of the chromosomes in the nuclei was either identical or formed a mirror image, suggesting that the position of the chromosomes in polarized nuclei may be constant. To confirm this observation, the angles formed by the two homologues in the polarized oval nuclei were measured for chromosomes 1, X, and 7. The measurements disclosed that, in about two-thirds of the nuclei, the two homologues formed angles of 150 degrees, 157 degrees and 148 degrees, nearly identical to those formed by the same three chromosomes in prometaphase rosettes of cultured diploid human fibroblasts (Nagele et al., 1995). In about one third of the nuclei, the same homologues formed angles of 89 degrees, 72 degrees, and 94 degrees, and occasionally an angle of 180 degrees. A three-dimensional computer reconstruction of the nuclei was performed using the data for the X chromosomes. By cinematographic technique, it could be documented that the angles separating the two homologues depended on the rotation of the nucleus along the axes X, Y, and Z. The cause of the rotation is speculative at this time. Because of the concordance of these data in terminally differentiated epithelial cells with prior observations on prometaphases of human diploid fibroblasts, it is suggested that the position of chromosomes in all human cells is constant throughout the cell cycle. The possible significance of these observations is discussed. PMID- 9858827 TI - Identification of the WBSCR9 gene, encoding a novel transcriptional regulator, in the Williams-Beuren syndrome deletion at 7q11.23. AB - We have identified a novel gene (WBSCR9) within the common Williams-Beuren syndrome (WBS) deletion by interspecies sequence conservation. The WBSCR9 gene encodes a roughly 7-kb transcript with an open reading frame of 1483 amino acids and a predicted protein product size of 170.8 kDa. WBSCR9 is comprised of at least 20 exons extending over 60 kb. The transcript is expressed ubiquitously throughout development and is subject to alternative splicing. Functional motifs identified by sequence homology searches include a bromodomain; a PHD, or C4HC3, finger; several putative nuclear localization signals; four nuclear receptor binding motifs; a polyglutamate stretch and two PEST sequences. Bromodomains, PHD motifs and nuclear receptor binding motifs are cardinal features of proteins that are involved in chromatin remodeling and modulation of transcription. Haploinsufficiency for WBSCR9 gene products may contribute to the complex phenotype of WBS by interacting with tissue-specific regulatory factors during development. PMID- 9858828 TI - Fluorescence in situ hybridization analysis of the replication properties of the myotonic dystrophy protein kinase (DMPK) gene region. AB - Myotonic dystrophy (DM) is caused by an expansion of a CTG repeat sequence in the 3' noncoding region of a protein kinase gene (DMPK) at 19q13.3. We used in situ hybridization to analyse the replication timing of the genomic region containing DMPK in fibroblasts and myoblasts from controls and myotonic dystrophy patients. In this method the relative proportion of singlet to doublet hybridization signals is used to infer the relative time of replication of specific loci or regions. Our results show that in cells from normal individuals approximately 65% of signals appear as doublets, indicating early replication. In DM patients with a number of CTG repeats ranging from about 600-1800 we observed a significant increase of singlet-doublets compared to the background level. These results suggest the existence of replication alternations and/or structural differences between the normal and mutant alleles induced by the presence of the DM mutation. PMID- 9858829 TI - Application of FISH for in situ detection and quantification of DNA breakage. AB - We describe a simple procedure that allows the use of fluorescence in situ hybridization (FISH) for in situ detection of DNA strand breaks in single cells (DBD-FISH: DNA Breakage Detection-FISH). After trapping within an agarose microgel, cells are incubated in an unwinding alkaline solution, deproteinized and dehydrated. Areas of single-stranded DNA are generated by the alkaline solution in proportion to the degree of DNA strand breakage. These then act as targets for FISH of whole genomic or region-specific probes (telomeric, human chromosome 8 painting, human alphoid DXZ1 locus, and human c-erbB-2 cosmid probes). Measurement of the amount and surface of FISH signals provides information on the breakage level in probed areas, permitting the assessment of possible intragenomic differences in sensitivity as well as intercellular heterogeneity in DNA damage induction or repair. PMID- 9858830 TI - Identification of the sex chromosomes of the medaka, Oryzias latipes, by fluorescence in situ hybridization. AB - In the medaka, Oryzias latipes, which does not have cytologically recognizable sex chromosomes, the sex is genetically determined and the mechanism of sex determination (XX/XY) can be revealed by genetic crosses using a particular pigment gene. In a previous study, we isolated a sex-linked DNA marker (SL1) using the genomic differences between inbred strains of medaka. In the present paper, we further isolated another sex-linked clone (pHO5.110). The pHO5. 110 related sequences were tightly linked to sex in O. latipes. We designated the locus of the pHO5.110-related sequence on sex chromosomes of medaka as Sex-Linked 2 (SL2). Southern blot analyses suggested that the pHO5.110-related sequence was tandemly repetitive in the medaka genome. Using the clone as a probe for FISH analysis, strong hybridization signals were obtained in a couple of chromosomes that formed one of two large submetacentric chromosome pairs. The pHO5.110 related sequences were repetitive in the genomes of other species of Oryzias (O. curvinotus, O. luzonensis and O. mekongensis) that are karyologically related to O. latipes (all are members of the so-called biarmed group). By contrast, the sequences were not detected as repetitive in other Oryzias species. Hence, it is thought that pHO5.110-related sequences were amplified in the genome of a common ancestor of the biarmed group. PMID- 9858831 TI - Karyotype evolution of marsupials: from higher to lower diploid numbers. AB - A basic 2n = 14 ancestral marsupial karyotype giving rise to higher diploid numbers through chromosome fissions has been widely accepted for the last three decades. Our finding of interstitial telomeres in two South American species, one with the 2n = 14 "ancestral karyotype" and the other with 2n = 18, indicates that these complements evolved from a karyotype with a higher diploid number. A new scenario for the karyotype evolution in the group is put forward. In this scenario an ancestral karyotype with at least 22 chromosomes would have originated the basic karyotype with 2n = 14 before the radiation of marsupials. PMID- 9858832 TI - Assignment of GALGT encoding beta-1, 4N-acetylgalactosaminyl-transferase (GalNAc T) and KIF5A encoding neuronal kinesin (D12S1889) to human chromosome band 12q13 by assignment to ICI YAC 26EG10 and in situ hybridization. medjph@stjames.leeds.ac.uk. PMID- 9858833 TI - Assignment of the fetoprotein transcription factor gene (FTF) to human chromosome band 1q32.11 by in situ hybridization. PMID- 9858834 TI - Assignment of the human BC200 RNA gene (BCYRN1) to chromosome 2p16 by radiation hybrid mapping. PMID- 9858835 TI - A serpin gene cluster on human chromosome 6p25 contains PI6, PI9 and ELANH2 which have a common structure almost identical to the 18q21 ovalbumin serpin genes. AB - The human genes encoding the "ovalbumin" subgroup of closely related serine proteinase inhibitors (serpins) are located at 18q21.3 and 6p25. Those at 6p25 include proteinase inhibitor 6 (PI-6; gene symbol PI6), proteinase inhibitor 9 (PI-9; gene symbol PI9) and monocyte neutrophil elastase inhibitor (M/NEI; gene symbol ELANH2). Here we describe the fine mapping of these genes to a 200-kb region of chromosome 6 that includes the markers WI-8835 and D6S1338, and the establishment of the gene order: tel-PI6-PI9-ELANH2-cen. PI6 and ELANH2 are transcribed towards the telomere, and structural analysis shows that PI6 and PI9 are organized identically, having seven exons and six introns. PI6 and PI9 are almost identical in structure to the ovalbumin serpin genes at 18q21.3. The 18q21.3 genes have an extra exon and intron, otherwise all the other exon/intron boundaries are conserved between the two groups. These results represent the first detailed map of the chromosome 6 serpin gene cluster, and demonstrate that although they are very closely related, the 6p25 and 18q21-->q23 ovalbumin serpin genes form two structurally distinct groups. These findings do not support a previously proposed model for evolution of the clusters which invoked an inter chromosomal duplication of the entire 6p25 group to 18q21.3. PMID- 9858836 TI - The genomic breakpoint and chimeric transcripts in the EWSR1-ETV4/E1AF gene fusion in Ewing sarcoma. AB - Chromosome translocation creates a fusion between the EWSR1 gene and an ETS family gene. The fusion between these two genes is a characteristic feature of Ewing sarcoma. We previously identified a fourth translocation, t(17;22)(q12;q12), in genomic DNA isolated from cells of patients affected with Ewing sarcoma. The discovery of this translocation suggested that there might be a novel EWSR1-ETV4 fusion gene. In the present study, we determined the genomic breakpoint and characterized the chimeric transcript of the EWSR1-ETV4 fusion gene in two t(17;22) Ewing sarcomas. Reverse transcriptase-PCR assay showed an in frame fusion between the 5'-terminal region of EWSR1 and the 3' end of ETV4 (alias E1AF, PEA3); the chimeric transcript could thus serve as a template for expression of a protein composed of the N-terminal portion of EWSR1 fused to the DNA-binding domain of ETV4. Long PCR and sequence analysis of genomic DNA revealed that either exon 8 or intron 7 of EWSR1 is fused to the same intron of ETV4 in both tumors. Several palindromic oligomer sequences were found close to the breakpoints in both genes. The 159-bp Alu-like sequence was repeated in the breakpoint region of the ETV4 gene. These observations suggest a mechanism of EWSR1-ETV4 gene fusion. PMID- 9858837 TI - Comparative genomic hybridization and its application to Wilms' tumorigenesis. AB - Eighty sporadic Wilms' tumor samples were analyzed by comparative genomic hybridization (CGH) to identify chromosomal regions involved in the etiology of the disease. Twenty percent of the samples showed chromosomal gains or losses. The majority of chromosomal gains and losses were similar to those identified through molecular and cytogenetic studies. Gains were observed on chromosomes 1q, 7q, 8, and 12, whereas losses were found on chromosomes 1p, 4p, 4q, 7p, 16q, 18q, 21q, and 22q. Other genetic aberrations identified in this study included deletions of chromosomes 5p and 15q, as well as gains of discrete loci on chromosomes 3p and 3q. These latter regions have not been previously implicated in Wilms' tumorigenesis and may contain novel genes relevant to the development and/or progression of this disease. PMID- 9858838 TI - Neuronal migration and differentiation in the development of the mouse dorsal cochlear nucleus. AB - The dorsal cochlear nucleus (DCN) of mammals displays a cortical structure containing a number of cell types organized into distinct layers. In the present study, the migratory mode of large multipolar cells and granule cells as well as the morphological differentiation of the projection neurons were investigated in the development of the mouse DCN. The classification of the DCN neurons followed that of Ryugo and Willard. The mode of neuronal migration was examined by immunohistochemical bromodeoxyuridine labeling. Large multipolar neurons originated from the primary rhombic lip and small granule cells from the secondary rhombic lip. Large multipolar neurons migrated radially from the ventricular zone into the forming DCN. Granule cells were generated later than the large multipolar neurons and migrated via the subependymal and subpial routes. Large multipolar neurons and small granule cells were thus segregated early in the DCN development and intermixed later during perinatal maturation. Projection neurons retrogradely labeled by DiI application to the contralateral inferior colliculus showed neurite extension between the pial surface and the ventricular zone during migration in the DCN primordium. The retrogradely labeled projection neurons showed a well-differentiated morphology of the large multipolar neurons as early as the late embryonic stage. The arrangement of the radial glial processes coincided with that of the migratory projection neurons. The migratory immature neurons showed close apposition with the radial glial processes, suggesting that glial scaffolds are involved in the migration and settlement of the large multipolar neurons. Thus, it is suggested that the mode of migration and settlement of large multipolar neurons and granule cells in the developing DCN is highly similar to that of Purkinje and granule cell migration in the cerebellar development, based on the findings of this study and the structural similarity between the cerebellum and DCN. PMID- 9858839 TI - Proliferation of human Schwann cells induced by neu differentiation factor isoforms. AB - Neu differentiation factor (NDF), a 44-kD polypeptide, is a member of the neuregulin family which also includes glial growth factor, heregulin and acetylcholine-receptor-inducing activity. Previous studies have demonstrated that NDF/glial growth factor/heregulin/acetylcholine-receptor-including activity are products of neurons and mediate proliferation, differentiation and gene expression in Schwann cells of experimental animals. In the present study, the efficacy of different isoforms of NDF in stimulating human Schwann cell proliferation is investigated in Schwann-cell-enriched cultures derived from fetal human dorsal root ganglia (15-20 weeks gestation). NDF isoforms examined include alpha1, alpha2, EGF-like domain alpha2 (EGFalpha2), alpha3, beta1, EGFbeta1, EGFbeta, beta2 and beta3. For the assessment of Schwann cell proliferation, double immunostaining using antibodies specific for S-100 protein and bromodeoxyuridine was used. While treatment of Schwann cells with NDF alpha isoforms (alpha1, alpha2, alpha3 and EGFalpha2) had little effect on Schwann cell proliferation, NDF beta isoforms (beta1, beta2, beta3, EGFbeta1 and EGFbeta) induced a greatly enhanced proliferation in Schwann cells. The proliferation index in unstimulated Schwann cells was 1.3 +/- 0.9%, whereas in Schwann cells treated with NDFbeta isoforms the proliferation index was 21.8 +/- 2.2%. The finding that the truncated beta isoforms such as EGFbeta1 and EGFbeta retain a mitogenic activity as potent as full-length beta isoform indicates that the C terminal portion of the EGF-like domain is responsible for its receptor binding and subsequent biological activity. PMID- 9858840 TI - Immunoultrastructural expression of ICAM-1 and PECAM-1 occurs prior to structural maturity of the murine blood-brain barrier. AB - This study investigated the time of expression of two endothelial cell adhesion molecules, ICAM-1/CD54 and PECAM-1/CD31 in the developing postpartum mouse blood brain barrier (BBB). Immunoultrastructural studies demonstrated that both adhesion molecules are initially expressed primarily on the luminal endothelial cell surfaces at birth or shortly thereafter, they increase in an intensity at approximately 1 week postpartum and then decrease to a weak labeling of the luminal endothelial cell surfaces at 2 weeks after birth. In the adult animals, little or no adhesion molecule labeling was expressed on blood vessels. Our results present immunocytochemical evidence that upregulation of ICAM-1 and PECAM 1 occurs prior to structural maturity of the BBB. Moreover, these data support a notion that these adhesion molecules play an important role for angiogenesis in the developing murine BBB. PMID- 9858841 TI - Dose-dependent effects of multiple acute cocaine injections on maternal behavior and aggression in Sprague-Dawley rats. AB - Rat dams, which had no prior drug treatment, were either nontreated controls or were injected subcutaneously 4 times during a 10-day period with a single dose of 30, 15 or 7.5 mg/kg of cocaine hydrochloride HCl, or normal saline. Injections were given immediately postpartum following delivery of their final pup (PPD 1), and again on postpartum day 3 (PPD 3), postpartum day 6 (PPD 6) and postpartum day 10 (PPD 10). Dams were observed 30 min following injections for maternal behavior (MB) towards 8 surrogate male pups on PPD 1 and PPD 3 and for aggression towards a male or female intruder in the presence of their litter on PPD 6 and PPD 10. Compared to saline and untreated controls, cocaine-treated dams exhibited more disruptions in MB on both PPD 1 and PPD 3 and were less aggressive towards an intruder, regardless of intruder sex, on PPD 6 and PPD 10. In most cases MB was altered in a dose-dependent manner with the higher doses of cocaine resulting in a greater disruption of behavior. PMID- 9858842 TI - Myelin proteolipid protein: function in myelin structure is distinct from its role in oligodendrocyte development. AB - The myelin proteolipid proteins PLP and DM20 are essential for the compaction of central nervous system myelin and they play an important role in the maturation of the oligodendrocyte. The specific function of the less abundant DM20 isoform is still unknown, but rescue experiments previously indicated that both isoforms are necessary for oligodendrocyte maturation. In vitro experiments have suggested DM20 may assist in the translocation of PLP into the membrane. We tested this hypothesis in vivo, by investigating whether wild-type PLP derived from a transgene could be incorporated into the myelin membrane of Plp mutant rumpshaker mice. We previously demonstrated that expression of the PLP transgene alone in a more severe Plp mutant, jimpy mouse, did not result in PLP incorporation into the myelin. Here we report that there was significantly more PLP in white matter from rumpshaker expressing the PLP transgene than their nontransgenic rumpshaker littermates and that myelin structure was improved. The delay in oligodendrocyte development was not alleviated by expression of the PLP transgene however, supporting an essential role for DM20 in oligodendrocyte maturation. PMID- 9858843 TI - Enhanced epileptogenicity of area tempestas in the immature rat. AB - Age-related differences in kindling have been previously observed in terms of behavioral manifestations, seizure generalization and intensity of postictal refractory period. In the present study the development of kindling in the area tempestas (specifically the dorsal endopiriform nucleus) was examined in developing rats (15-16 days of age). Results indicate that kindling can be easily elicited from the immature area tempestas. Rats kindled from area tempestas reached the early kindling stages significantly faster, had longer afterdischarges and a greater number of severe stage 6 seizures than rats kindled from areas above the area tempestas or in the piriform cortex proper. This study indicates that the area tempestas is particularly epileptogenic in the immature rat. PMID- 9858844 TI - Motoneurones that innervate the rat soleus muscle mature later than those to the tibialis anterior and extensor digitorum longus muscles. AB - The response of motoneurones that innervate either the soleus or tibialis anterior (TA) and extensor digitorum longus (EDL) muscles to increased locomotor activity or to nerve injury at different stages after birth was examined. Increased locomotor activity of rat pups was induced by daily treatment with L dopa during the first 12 days after birth, and the number of surviving motoneurones to the soleus or TA/EDL muscles was established by retrograde labelling. Treatment with L-dopa resulted in the loss of a significant number of motoneurones within the soleus motor pool but had no effect on the survival of those motoneurones innervating the TA/EDL. Furthermore, following nerve injury during the first few days postnatally, more motoneurones within the soleus motor pool die than in the TA/EDL pool. These results indicate that motoneurones to the soleus muscle mature later than those to the TA/EDL muscles. PMID- 9858845 TI - Myelin proteolipid protein intron 1 sequences do not appear to enhance myelin proteolipid protein gene transcription. AB - Sequences from the first intron of the mouse myelin proteolipid protein (PLP) gene were examined for their ability to modulate PLP gene expression. Glial (N20.1) or nonglial (NIH 3T3) cells were transiently transfected with constructs that contained 1.4 kb of PLP promoter sequence driving luciferase reporter gene expression, as well as various portions of PLP intron 1 DNA. Although these same PLP intron 1 fragments enhanced reporter gene expression from a heterologous basal promoter in a previous study, the results reported here demonstrate that they do not augment PLP promoter activity. Thus, the regulation of PLP cell-type specific expression, conferred by the first intron, appears to be mediated by an enhancer-independent mechanism. PMID- 9858846 TI - Effect of Leuprorelin Acetate on Cell Growth and Prostate-Specific Antigen Gene Expression in Human Prostatic Cancer Cells. AB - Objectives: We investigated modulation of cell growth and prostate-specific antigen (PSA) gene expression in prostatic cancer cells by the luteinizing hormone-releasing hormone analog (LH-RHa), leuprorelin acetate, alone or combined with other agents. Methods: The effect of the analog on proliferation of both androgen-sensitive and -insensitive prostate cancer cells, maintained in different culture conditions, was evaluated by cell counts at various intervals of time. Basal expression of PSA gene and its variations were determined by a reverse transcriptase-polymerase chain reaction assay. Results: LH-RHa is ineffective in regulating cell growth, when used alone in both hormone-sensitive and -insensitive cell lines. Nevertheless, it counteracts the stimulatory action of androgens on proliferation of LNCaP cells, which respond to low concentrations of dihydrotestosterone. Moreover, LH-RHa has an inhibitory effect on the mitogenic action of epidermal growth factor (EGF) in androgen-unresponsive PC-3 cells. The analog reduces PSA gene expression in both hormone-sensitive and insensitive cells. Interestingly, it counteracts the gene expression induced by androgens in LNCaP cells and by EGF in PC-3 cells. Conclusions: These data show that LH-RHa may behave like a negative growth factor, which directly regulates cell growth and PSA gene expression. Moreover, our findings support the idea that growth factors may interfere with the androgen signalling pathway. PMID- 9858847 TI - Therapeutic Options in Locally Defined or Advanced Prostate Cancer. AB - Objective: Critical discussion of the available therapeutic options in locally defined or advanced prostate cancer (PCa) in the absence of standards or guidelines. Methods: Medline-based survey of the pertinent literature. The therapeutic options are presented stage-by-stage. Results: Intracapsular PCa (T1a T2bN0M0) is preferably managed by radical prostatectomy; 'insignificant' cancers may be treated expectantly. The outcome of irradiation is not as predictable as radical surgery. The following points regarding treatment of locally advanced PCa are still under debate: neoadjuvant treatment is probably not as efficient as believed; technical refinements of radical prostatectomy may be helpful; in some instances, the value of adjuvant treatment is undecided. The efficacy of irradiation in conjunction with androgen deprivation probably equals the efficacy of surgery. No consensus can be found in the presence of positive lymph nodes, although single institutions achieve remarkable results with surgery plus androgen deprivation. Conclusion: Prognostic factors beside the clinical stages and grades should be incorporated in the treatment decision, and quality-of-life measurement should gain more importance when the treatment outcome is assessed. PMID- 9858848 TI - Adjuvant Hormonal Treatment with Radiotherapy for Locally Advanced Prostate Cancer. AB - Objectives: Long-term results of radiotherapy in locally advanced prostate cancer are poor due to local and distant failures. Since prostate cancer is hormone dependent, tumor androgen deprivation may enhance tumor eradication. Methods: Three randomized phase III trials, RTOG and EORTC are reported: they assess androgen suppression by using a luteinizing hormone-releasing hormone analogue (LH-RHa) with or without androgen blockade before and during, or during and after external irradiation. Results: A gain in disease-free, local relapse-free and metastasis-free survival has been obtained (p < 0.001). Only the EORTC 22863 trial has reported a significant improvement in overall survival (p = 0.001) with an LH-RHa started the first day of radiotherapy and administered every 4 weeks over 3 years. In the RTOG 85-10 trial, and LH-RHa, initiated in the last week of radiation therapy and continued until relapse, increased overall survival only in patients with poorly differentiated tumor with a Gleason score of 8-10 (p = 0.03). Conclusion: Androgen suppression prior to and during radiation improves disease-free survival; adjuvant hormonal therapy with an LH-RHa during and after radiation improves overall survival. PMID- 9858849 TI - Intermittent Complete Androgen Blockade in PSA Relapse after Radical Prostatectomy and Incidental Prostate Cancer. AB - Objectives: To determine the efficacy, safety and feasibility of intermittent androgen deprivation (IAD) in patients with prostate-specific antigen (PSA) relapse after radical prostatectomy or with an incidental prostate cancer (pT1B) after transurethral resection of the prostate (TURP). Methods: Open, nonrandomized, prospective pilot study using the luteinizing hormone-releasing hormone analogue (LH-RHa), leuprorelin acetate (1-month depot) and cyproterone acetate. Results: Forty-four patients have been enrolled. After a 30-64 months' follow-up no progression to androgen-independent status has been observed. Of the entire observation period, 26.6 months (44-58%) remained treatment-free. During the treatment-free periods, normal testosterone levels were obtained, resulting in a cessation of the symptoms of androgen suppression and an improvement in quality of life. Conclusions: These results indicate that IAD is an effective and feasible therapy in patients with early stages of prostate cancer. Larger trials are necessary to confirm these encouraging results. Therefore, a European prospective, randomized, multicenter study (RELAPSE study) has been started to compare IAD with continuous androgen blockade in terms of time to tumor progression, safety and quality of life in patients with PSA relapse after radical prostatectomy. PMID- 9858850 TI - Intermittent Complete Androgen Blockade in Metastatic Prostate Cancer. AB - Introduction: Intermittent hormonal treatment of prostate cancer was first developed based upon experimental study results. Using the Shionogi mouse breast cancer model, it was shown that the tumor grows rapidly in the presence of androgens, then undergoes apoptotic regression when androgens are removed. This apoptotic potential can be reinduced several times by cyclic replacement and withdrawal of androgens. These results led to the concept being evaluated in clinical trials. Method: In most of the clinical studies a protocol is used in which the patient receives 36 weeks of androgen deprivation. For those patients whose prostate-specific antigen (PSA) drops to less than 4 ng/ml within 32 weeks of therapy, the androgen withdrawal is stopped at 36 weeks, and not reintroduced until PSA increases to 20 ng/ml. This cycle is then repeated until the patient's tumor becomes hormone-sensitive. Results: Akakura et al. in 1993, reported on 7 patients who had a total of 12 episodes off-hormone therapy after achieving PSA complex remission, with further response after each re-exposure. Bruchovsky et al. in 1997, reported on 47 patients who entered a study of intermittent hormone therapy to evaluate the effect of cyclic withdrawal and replacement therapy in 14 D2, 10 D, 19 C, 2 B2, 2 A2 patients. Treatment was initiated with combined androgen blockade and continued for at least 6 months until a serum PSA nadir was observed. The first two treatment cycles lasted 73 and 75 weeks, with a mean time off therapy of 30 and 33 weeks and an overall mean percentage time-off therapy of 41 and 45%. Serum testosterone returned to the normal range within 8 weeks (range: 1-26 weeks) of stopping treatment. The off-treatment period in both cycles was associated with an improvement in sense of well-being, and the recovery of libido and potency in the men who reported normal or near-normal sexual function before the start of therapy. In 7 patients with stage D2 disease, the cancer progressed to an androgen-independent state. The mean and median times to progression were 128 and 108 weeks. Seven patients died, one from a non-cancer related illness, with mean and median overall survival times of 210 and 166 weeks. Conclusion: These studies demonstrated that the androgen-dependent state of prostate cancer can be maintained during a course of intermittent androgen suppression, supporting the possibility of multiple apoptotic regressions under well-regulated conditions. Oliver et al. in 1997, conducted a retrospective study of 20 patients and concluded that intermittent androgen deprivation reduced induction of hormone-resistant prostate cancer, with no acute or major risk associated with the use of intermittent androgen suppression. More clinical studies are required to clarify the indication for intermittent hormone therapy and evaluate improvement in quality of life and survival. In the future, approaches to the improvement of therapeutic apoptosis could include intermittent hormone therapy, associated with additive cytotoxic therapeutic strategies. PMID- 9858851 TI - Analysis of three suggested psoriasis susceptibility loci in a large Swedish set of families: confirmation of linkage to chromosome 6p (HLA region), and to 17q, but not to 4q. AB - Psoriasis is known to be a heterogeneous disease with so far three reported major psoriasis susceptibility loci on chromosome 4q, 6p and 17q. In this study we investigated three reported gene locations by nonparametric and parametric linkage analysis in a large family set consisting of 104 families (153 sib pairs) from Sweden. We could confirm linkage to chromosome 6p. A maximum heterogeneous lod score of 2.78 was reached at locus D6S276 (alpha = 0.60). Allelic association studies within the HLA region indicated linkage disequilibrium at locus TNFbeta with a significant p value of 0.0009. Furthermore, we obtained weak evidence of linkage to the locus on chromosome 17q while no evidence of linkage could be found to the chromosome 4q region. PMID- 9858852 TI - Allele frequencies in a worldwide survey of a CA repeat in the first intron of the CFTR gene. AB - A dinucleotide CA repeat within intron 1 of the CFTR gene has recently been identified. We have determined the allele frequencies of this polymorphism in samples from 18 populations covering all major geographical areas, with a total of 1,816 chromosomes. When considering allele distributions, African populations presented a wider range of alleles than other populations and also presented higher expected heterozygosities. Analysis of molecular variance showed that 8.04% of the genetic variance in this locus could be attributed to differences among populations. We concluded that the polymorphism in the CA repeat in intron 1 of the CFTR gene is highly informative in populations from all geographical regions of the world. Thus, it can be applied to family studies of unknown mutations causing cystic fibrosis (CF) and can provide valuable information for genetic counseling. Moreover, its analysis should be included in the haplotypic analysis of known CF mutations. PMID- 9858853 TI - Polymorphisms in the HFE gene. AB - Hereditary hemochromatosis is an autosomal recessive disease characterized by progressive iron overload. Recently, a candidate gene named HFE was isolated on the short arm of the chromosome 6 within which two mutations were identified: C282Y and H63D. To date, only homozygosity for the C282Y mutation is considered as a diagnostic criterion of hemochromatosis. 7.6% of the patients studied in our laboratory did not carry two copies of the C282Y mutation. On the other hand, a dysmetabolic iron overload syndrome has recently been described and the search for the C282Y and H63D mutations revealed that none of the patients was homozygous for C282Y while 67% exhibited one of the mutations. The possibility of a new mutation in the HFE gene has been raised to explain the disease in the remaining patients, as well as, in the few hemochromatotic patients without two copies of the C282Y mutation. The aim of this study was to search for new mutations in the HFE gene in 16 such patients. Direct sequencing of exons and 3 introns did not reveal any new mutation but identified a few polymorphisms. PMID- 9858854 TI - Determination of the allele frequencies of three polymorphisms in the promoter region of the human protein C gene in three Brazilian ethnic groups. AB - The frequencies of three polymorphisms in the promoter region of the human protein C gene have been determined in three ethnic groups of the Brazilian population. The allele frequencies observed in south-eastern Brazilian Caucasians and Blacks were similar to the values obtained for the Dutch population and were different from those observed in Amazonian Indians living in the north of the country. The most frequent genotypic combination found in this latter group was the CC/GG/AA genotype, which is very rare among Caucasians. The complete heterozygous genotype CT/AG/AT was the most frequent both in Brazilian Caucasians and Blacks. Among 27 possible genotypic combinations, 21 were found in the Caucasian group and 15 in Blacks, revealing a high degree of genetic diversity in south-eastern Brazilian populations. PMID- 9858855 TI - Apolipoprotein B 5'-Ins/Del and 3'-VNTR polymorphisms in Chinese, malay and Indian singaporeans. AB - The allele frequencies for the apolipoprotein B (apo B) 5'-Ins/Del and 3'-VNTR polymorphisms varied significantly (p < 0.01) among Singaporeans of Chinese, Malay and Indian descent. We calculated the unbiased expected heterozygosities for the 5'-Ins/Del polymorphism as 0.3357, 0.1984 and 0.2418, and for the 3'-VNTR as 0.5980, 0.5260 and 0.6749, respectively, in the Chinese, Malays and Indians. Compared to heterozygosities reported for other populations, the Singaporeans differed from most Caucasians in having significantly lower values but were closely related to other non-Caucasians. Thirteen alleles, with a bimodal distribution, were observed at the 3'-VNTR polymorphic locus; the alleles occurring most frequently among the Chinese and Malays were of 35 or 53 repeats, and among the Indians, of 37 or 47 repeats. The Del allele was associated with elevated serum cholesterol (p = 0.023), LDL-cholesterol (LDL-C) (p = 0.001) in the Chinese, and apo B (p = 0.007) in the Indians. Likewise, the larger 3'-VNTR alleles (> 41 repeats) were associated with raised cholesterol (p = 0.018), LDL-C (p = 0.025), and triglyceride (p = 0.001) in the Chinese. The two polymorphisms were not in significant linkage disequilibrium (D = -0.0029, p = 0.494) in the three ethnic groups. PMID- 9858856 TI - Population study of common glucose-6-phosphate dehydrogenase mutations in Kuwait. AB - DNA samples from 206 unrelated Kuwaitis of both sexes, i.e. 200 randomly selected individuals and 6 glucose-6-phosphate-dehydrogenase (G6PD)-deficient probands, have been analyzed by the PCR/RFLP technique for mutations underlying the most common G6PD-deficient variants (Mediterranean and A-). At the first step all samples were studied for the 563C-->T and 376A-->G mutations, then the samples positive for 376A-->G were further analyzed for 202G-->A, 680G-->T and 968T-->C mutations. Three mutations (563C-->T, 376A-->G and 202G-->A) were found to be present in the Kuwaiti population at polymorphic frequencies (0.0503, 0.0215 and 0.0111, respectively). Nineteen out of 20 unrelated Kuwaiti chromosomes with 563C ->T had Mediterranean haplotype as judged by 1311C-->T polymorphism. The frequency of G6PD-deficient genotypes was 4.5% (5.73% in males and 2.56% in females). PMID- 9858857 TI - Variable distribution of TFF2 (Spasmolysin) alleles in Europeans does not indicate predisposition to gastric cancer. AB - Peptides belonging to the trefoil factor family (TFF) protect the gastrointestinal epithelia. Overexpression of TFFs was observed in pathological conditions such as gastritis, ulceration, metaplasia and neoplasia of the gastrointestinal tract. The aims of this work were to investigate the recently described TFF2 gene polymorphism in different European populations. DNA samples from blood of healthy individuals and gastric cancer patients were genotyped using the polymerase chain reaction. They were compared to a gastric cancer population. The results do not show any significant difference in allelic frequencies between gastric cancer patients and healthy individuals from Portugal. However, the frequency of the two alleles found varies considerably among Europeans. PMID- 9858858 TI - Genomic distribution and gonadal mRNA expression of two human luteinizing hormone receptor exon 1 sequences in random populations. AB - Exon 1 of the human luteinizing hormone receptor (LHR) gene coding region exhibits at least two forms of sequence heterogeneity between 37 and 60 bp, spanning the junction of the signal peptide and the amino terminus of the mature protein. The LHR 1 differs from the LHR 2 by the insertion of 6 bp in exon 1 but is of identical sequence in the 5' flanking region. RFLP analysis of the two haplotypes within a random population of 63 individuals revealed allele frequencies of 0. 37 and 0.63 for LHR 1 and LHR 2, respectively. 94% of the samples contained at least one LHR 2 allele, whereas only 68% contained the LHR 1 allele. No gender differences were observed, and both homozygotes and heterozygotes displayed apparently normal reproduction. Reverse-transcriptase polymerase chain-reaction analyses of LHR mRNA from testes and ovaries revealed that both haplotypes are transcribed in normal individuals, with no difference in tissue specific distribution. Thus, at least two functional polymorphic forms of exon 1 coding region of the same LHR gene are present in a random human population. PMID- 9858859 TI - alpha1-antitrypsin (PI) alleles as markers of Westeuropean influence in the Baltic Sea region. AB - The distribution of alpha1-antitrypsin (PI) alleles was studied in an attempt to elucidate migrations and admixture between populations in the Baltic Sea region. The frequency of the PI Z allele, a typically Northwesteuropean marker gene, showed a highly significant regional variation in the Baltic Sea region. The highest frequency (4.5%) was found in the western part of Latvia (Courland). The PI S allele, another marker of Westeuropean influence, also showed an increased frequency in the Courland population. These results indicate that among the populations east of the Baltic Sea the Curonian population has the most pronounced Westeuropean influence. Archaeological data have shown that from the 7th century and for several hundreds of years Courland received immigrations from mainland Sweden and the island of Gotland. We speculate that the increased frequencies of the PI Z alleles and S alleles in Courland may have been caused by these migrations. PMID- 9858860 TI - Identification in Portugal and Brazil of a mtDNA lineage containing a 9-bp triplication of the intergenic COII/tRNALys region. AB - Although the deletion of one of the 9-bp repeats in region V of mitochondrial DNA is very common in Asians, Asian-derived populations and Africans, the triplication of the 9-bp segment was described only a few times, mostly on individuals from Asian origin. Here, we report for the first time the presence of the 9-bp triplication in Europeans. The triplication was initially found in one Brazilian individual. Sequencing of the hypervariable segments I (HVSI) and II (HVS2) of the control region and RFLP analysis of the coding region classified the mtDNA as belonging to the European haplogroup H. Since white Brazilians are predominantly of Portuguese descent, we screened 96 unrelated Northern Portuguese for the 9-bp triplication and found its presence in two of them (2.1%). One of these had an mtDNA haplotype identical to that of the Brazilian individual, while the other differed in a single base change in HVS2. The fact that the 9-bp triplication has reached polymorphic frequencies in Northern Portugal and that it has apparently differentiated into at least two lineages defined by the mutuation in HVS2 suggests that it probably occurred a long time ago. PMID- 9858861 TI - A novel mutation (R271X) in the myotubularin gene causes a severe miotubular myopathy. AB - The mutation is a C to T transition at nucleotide 811 of the MTM1 gene (OMIM 310400) leading to premature termination of translation at codon 271 of the myotubularin protein (R271X). PMID- 9858862 TI - Allele frequency of D17S855 microsatellite locus in Japanese people. AB - Allele frequency of D17S855 microsatellite locus, an intragenic polymorphic cytosine-adenine repeat located in the BRCA1 gene, was analyzed in 456 normal tissues of Japanese adults. The frequency of D17S855 microsatellite polymorphism was 77.9% (335/456). The size of D17S855 PCR fragments ranged from 137 to 157 bp. The most frequent allele was 151 bp (29.9%) followed by 149-bp (19.3%), 145-bp (17.5%), and 147-bp (17.0%) fragments. The observed allele distribution of this microsatellite was similar to that of Southern Swedes, while it was quite different from that of European Caucasian specimens deposited in the Genome Database. PMID- 9858863 TI - Absence of the derived allele at the DYS199 locus in 3 NE Chinese populations. AB - The allelic states of DYS199 locus were studied by allele-specific PCR. We found that all of the 107 samples chosen from three populations of northeast China had DYS199 C alleles. The result excluded the possibility that this mutation first identified in native American populations came from the three populations of northeast China. PMID- 9858864 TI - Parathyroid hormone and calcitriol in hypertension caused by dietary calcium deficiency in rats. AB - Hypertension caused by calcium deficiency in the diet has been linked with an increase in parathyroid hormone (PTH) and calcitriol levels. We evaluated arterial blood pressure (ABP), PTH, and calcitriol in normotensive Sprague-Dawley rats (SDR) and in spontaneously hypertensive rats (SHR) fed from weaning on a control diet with a normal calcium content (1%) or a low-calcium diet (0.1%). The calcemia was also measured in the rats by colorimetric methods. The low-calcium diet decreased the calcemia in both strains and brought about an increase in the ABP which was significant in adult SDR and particularly noticeable during the early hypertensive phase in SHR. The rats fed on this diet had higher hormonal plasma levels when compared with the corresponding values in rats fed on the control diet. In particular, the SDR fed on the low-calcium diet showed much higher PTH (122.6 +/- 31.0 pg/ml, p or = 10 mM concentration, inhibited proliferation and reduced the viability of human pancreatic cancer (Panc-1) cells. The drug concentration determined the extent of inhibition, and with continued culture a proportion of the cells detached, most of which stained with trypan blue. Although hypodiploid cells were detected by flow cytometry of cells cultured with 20 mM NTBN, DNA laddering was absent and the TUNEL reaction negative. "Dark" cells present in samples cultured with 10 mM NTBN exhibited decreased cytoplasmic volume and increased staining with methylene blue and azure II, but lacked characteristic nuclear changes of type 1 programmed cell death. Cells cultured with > 10 mM of the spin trap exhibited nuclear and cytoplasmic changes more consistent with a non-type 1, type 2 variant of PCD with extensive cytoplasmic vacuolization. Careful analysis revealed evidence of marked pinocytosis in some cells. In view of the spin-trap associated pinocytosis, augmented uptake of chemotherapy in affected cells might be anticipated, but additive, synergistic or antagonistic interactions between NTBN and 5 fluorouracil were not observed. PMID- 9858886 TI - Gene therapy of gliomas: receptor and transcriptional targeting. AB - Through incremental increases in the overall therapeutic ratio of combined modality regimens, each addition of unique selective toxicity to a tumor moves one step closer to a cure. The primary advantage of adding gene therapy strategies to current oncologic regimens is the ability to design multiple levels of unique biologic selectivity into vectors using recombinant technology. This article presents an overview of current and potential methods for designing vectors targeted to high grade gliomas through selective cell entry or transcriptional regulation. Cell entry based methodologies are founded on increasing relative uptake of the vector through the chemical or recombinant addition of epitopes which bind to receptors selectively expressed on target cells. Transcriptional targeting utilizes promoter and enhancer systems which have potential for selectively activating transcription for transgene expression or vector propagation in target cells. PMID- 9858887 TI - Contradiction between the expected role of tumour suppressor genes and the evolutionist theory. PMID- 9858888 TI - Role of folic acid in modulating the toxicity and efficacy of the multitargeted antifolate, LY231514. AB - We studied the effects of folic acid on modulating the toxicity and antitumor efficacy of LY231514. Using several human tumor cell lines adapted to growth in low folate medium, folic acid was shown to be 100- to 1000-fold less active than folinic acid at protecting cells from LY231514-induced cytotoxicity. The lethality of LY231514 was compared in mice maintained on standard diet or low folate diet. The LD50 occurred at 60- and 250-fold lower doses of LY231514 in DBA/2 and CD1 nu/nu mice, respectively, maintained on low folate diet compared to standard diet. The L5178Y/TK-/HX-murine lymphoma was much more sensitive to the antitumor action of LY231514 compared to wild type L5178Y-S tumors. For mice on low folate diet, LY231514 at 0.3 and 1 mg/kg (qd x 10, i.p.) produced 100% inhibition of L5178Y/TK-/HX-lymphoma growth, and significant lethality occurred at > or = 3 mg/kg. For mice on standard diet, LY231514 produced > 95% inhibition of tumor growth at 30 to 300 mg/kg, but all mice died at 800 mg/kg. Folic acid supplementation was demonstrated to preserve the antitumor activity of LY231514 while reducing toxicity. The combination of folic acid with LY231514 may provide a mechanism for enhanced clinical antitumor selectivity. PMID- 9858889 TI - Viral and non-viral vectors for cancer gene therapy. AB - BACKGROUND: Our research has focused on developing improved delivery vectors for treating cancer by gene therapy using the tumor suppressor p53 gene. MATERIALS AND METHODS: Recombinant viral and non-viral vectors were used to deliver the p53 gene into non-small cell lung cancer (NSCLC) cells either in culture or as a subcutaneous tumor. Transduction of tumor cells was measured by beta-gal expression while tumor cell proliferation was used to measure the effect of p53. RESULTS: High level transduction was obtained in vitro and in vivo with a recombinant adenoviral vector, resulting in tumor cell growth inhibition in both models. A targeted, non-viral gene delivery vector based on the use of an EGF/DNA polyplex also resulted in efficient (as high as 66% transduction) and specific gene delivery in vitro when replication defective adenovirus was used as an endosome release agent. CONCLUSION: These vectors now provide improved methods to deliver therapeutic genes for cancer treatment by gene therapy. PMID- 9858890 TI - Mistletoe in immunology and the clinic (short review). AB - The recent meeting of the AGMIF (Working Group for Mistletoe Therapy and Immunological Research) held in Herdecke, Germany, on October, 3rd, 1997, covered recent developments in the field of immunological and biological properties of Viscum album L., the European mistletoe, which is used for adjuvant cancer treatment. So far, one extract component, the mistletoe lectin (ML)-1, was propagated by some researchers to be the only relevant substance within the extracts. However, immunological activities of other extract components such as polysaccharides, vesicles, chitin-binding lectin and their interactions discussed in the first part, underline the significance of the other components as well. In the second part, clinical evidence for the beneficial effects of subcutaneous and intratumoral application of mistletoe therapy was presented by different working groups. However, further research is of great importance to carefully analyse and characterize the involved molecules and exact mechanisms underlying the beneficial effects reported in this meeting. PMID- 9858891 TI - Effects of hydroxyapatite on spontaneous mammary tumourigenesis and related parameters in SHN virgin mice. AB - Spontaneous mammary tumourigenesis in a high mammary tumour strain of SHN virgin mice was inhibited by the chronic ingestion of a diet containing hydroxyapatite [HAP: Ca5(OH)(PO4)3] at 2.5% or 5% up to 7 months of treatments, while not thereafter. The decrease in the excretion of urinary component levels associated with mammary tumour development was prevented in the experimental group given HAP, which resulted in the higher component levels in the experimental group than in the control of the tumourous mice. The treatment little affected body weight change, the pattern of the oestrous cycle, endocrine organ weights, glucose tolerance, serum levels of glucose and free fatty acid and serum superoxide dismutase activity. The results suggest that the inhibition by HAP of mammary tumourigenesis is at least partly ascribed to its prevention of a decreased metabolic turnover as reflected by the higher excretion of urinary components in tumourous mice given HAP. PMID- 9858892 TI - The inhibitory effect of adenovirus-mediated p16INK4a gene transfer on the proliferation of lung cancer cell line. AB - Abnormalities in the p16INK4a tumor suppressor gene are found in many lung cancer cell lines and primary lung cancer tissue. To examine its tumor suppressor function and potential adequacy in cancer gene replacement therapy, wild-type p16INK4a gene was inserted in an adenovirus derived gene delivery system and introduced into lung cancer cell lines (NCI-H441 and NCI-H157) that did not express p16INK4a. Western blot assay and immunocytochemistry demonstrated production of wild-type p16 protein in these cell lines. The biological function of exogenous p16 protein was confirmed by the inhibition of pRB phosphorylation. The expression of exogenous p16 protein via recombinant adenovirus significantly inhibited cancer cell growth and colony formation in vitro of NSCLC that can not express endogenous p16. The flow cytometric analysis showed these results correlated with G1 cell cycle arrest. These observations suggest the value of adenovirally-mediated p16INK4a gene replacement therapy for lung cancer. PMID- 9858893 TI - Protective action of plant polyphenols on radiation-induced chromatid breaks in cultured human cells. AB - The present study was performed to determine whether plant polyphenols can protect human cells against radiation-induced DNA damage manifested as chromatid breaks. Since each chromatid contains a single continuous molecule of double stranded DNA, chromatid breaks represent unrepaired DNA strand breaks. The addition of green or black tea extracts, their polyphenols or curcumin to cultures of human skin fibroblasts or PHA-stimulated blood lymphocytes significantly reduced the frequencies of radiation-induced chromatid breaks. An exception to this general finding was that the green tea polyphenol, ( )epigallocatechin gallate, had no effect. The protective action of these plant polyphenols seems to result from their known antioxidant properties, particularly the scavaging of hydroxyl free radicals. Frequencies of chromatid breaks in cells arrested immediately after irradiation or 0.5 to 1.5 hours post-irradiation in the presence or absence of a DNA repair inhibitor, provide a measure of DNA damage. The results of the present study show that tea and other plant polyphenols can protect human cells against radiation-induced DNA damage. PMID- 9858894 TI - Pyruvate kinase and the interaction of amino acid and carbohydrate metabolism in solid tumors. AB - The interaction between glycolysis, glutaminolysis and tumor growth in WAG/Fra rnu/rnu rats has been investigated. Small tumors are characterized by a low conversion of glucose to lactate whereas the conversion of glutamine to lactate is high. In medium sized tumors the flow of glucose to lactate as well as oxygen utilization are increased whereas glutamine and serine consumption are reduced. At this stage the tumor cells start with glutamate and alanine production. Large tumors are characterized by a low oxygen and glucose supply but a high glucose and oxygen utilization rate. The conversion of glucose to glycine, alanine, glutamate, glutamine, and proline reaches high values and the amino acids are released. Pyruvate kinase increases with tumor weight and is positively correlated with an increase in glucose and oxygen utilization. The shift from glutamate consumption to glutamate production is correlated with an increase in glutamate dehydrogenase and glutamate oxaloacetate transaminase activity. PMID- 9858895 TI - Metabolic characteristics of different malignant cancer cell lines. AB - Extracellular AMP inhibits cell proliferation of MCF-7 and MDA-MB-453 whereas cell proliferation of the highly malignant Novikoff cell line is not affected. In medium with low glucose supply MDA-MB-453 cells grow well, Novikoff cells are slightly inhibited and MCF-7 cells are totally unable to grow. Isoelectric focusing revealed that a glyclytic enzyme complex exists in all three cell lines. In addition to the glycolytic enzymes, c-Raf-kinase, adenylate kinase, and nucleoside diphosphate kinase are also found within the complex. The differences in glucose in dependence of the three cell lines can be explained by the different constitutions of shuttle enzymes. MDA-MB-453 and Novikoff cells contain cytsolic glycerol 3-phosphate dehydrogenase which is associated with glyceraldehyde 3-phosphate dehydrogenase within the glycolytic enzyme complex and which is responsible for the transport of cytoslic hydrogen in the mitochondria. MCF-7 and Novikoff cells contain the pI 7.8 form of malate dehydrogenase which couples glycolysis with glutaminolysis. PMID- 9858897 TI - Assessment of apoptosis occurring in spleen cells from nitrogen mustard-treated or gamma-irradiated mice. AB - The short-term consequences on spleen cells of the intraperitoneal administration of nitrogen mustard (HN-2) to mice or of a whole-body gamma irradiation have been evaluated. Experiments were designed to assess the induction of apoptosis in spleen cells following exposure to these agents. The occurrence of this type of cell death was analysed by several methods, in particular the quantification in the blood of phosphotidylserine-bearing microparticles shed by apoptotic cells. In response to HN-2 or radiations, spleens undergo a rapid involution of their weight and cellularity. Ex vivo apoptosis occurs within 24 hours in cultured lymphocytes in a dose-dependent manner after both treatments. As compared with untreated controls, circulating microparticles increased 3-fold after the injection of 5 mg/kg of HN-2. PMID- 9858896 TI - Gene expression of cyclooxygenase-2, group II and cytosolic phospholipase A2 in human colorectal cancer. AB - The development of intestinal tumours in the Min mouse is influenced by Moml locus, where the group II phospholipase A2 (PLA2-II) gene has been suggested as the candidate gene. Since no causative genetic alterations have been identified in PLA2-II in human colorectal tumours, we tested whether PLA2-II or the cytosolic isoform cPLA2, are dysregulated in human colorectal carcinogenesis. By means of RT-PCR, the expression of these genes and cyclooxygenase-2 (COX-2), which is upregulated in colorectal tumours, were examined. No changes in gene expression of PLA2-II were found in 44 tumours compared to normal mucosa. cPLA2 and COX-2 were upregulated; however, no correlation in expression was found between these genes. Thus, the supply of arachidonic acid provided by PLA2 II/cPLA2 seems not to be the rate limiting step in PGE2 (a prostaglandin/mitogen) formation via COX-2 and PLA2-II plays a minor or no role in human colorectal carcinogenesis. PMID- 9858898 TI - Expression of somatostatin messenger RNA and receptor in cultured brain tumor cells. AB - In order to understand the role of somatostatin in tumor cell growth, the expression of somatostatin in 7 cultured brain tumor cells was investigated. Radioimmunoassay demonstrated the production of somatostatin into the culture supernatant of 6 tumor cells. The expression of somatostatin mRNA and receptor was investigated by reverse transcription polymerase chain reaction (RT-PCR) and a binding assay with 125I-somatostatin-14, respectively. Only two cell lines, GBS 1 and U87MG, expressed somatostatin mRNA, whereas somatostatin receptor was expressed in all cultured cells. These results suggest that somatostatin is an important factor for regulating cell growth. PMID- 9858900 TI - Doxorubicin cardiotoxicity and serum lipid increase is prevented by dextrazoxane (ICRF-187). AB - This study is related to the serious side effects of Doxorubicin-cardiotoxicity and serum lipid caused by the drug's cumulative effect. Studies were performed on experimental animals treated with intensive administration of Doxorubicin. Seventy five wistar rats were divided in two equal groups A and B. Group A was used for doxorubicin administration and B for doxorubicin and dextrazoxane. The drugs were administered weekly for twelve weeks at doses 0.2 mg/100 g BW for doxorubicin and 1.5 mg/100 g BW for dextrazoxane. Histological examination of the cardiac muscle, large vessels, liver and other organs and biochemical examination for serum lipids and liver enzymes were performed on certain weeks. Comparison of the findings of the two groups showed a) a reduction in doxorubicin cardiotoxicity by dextrazoxane and b) the addition of dextrazoxane to doxorubicin resulted in lowering the increase of serum lipids produced by doxorubicin. c) In vitro tests by chemiluminescence showed that dextrazoxane acts as a scavenger of oxygen free radicals. PMID- 9858899 TI - Interleukin-1 alpha and basic fibroblast growth factor induction of matrix metalloproteinases and their inhibitors in osteosarcoma cells is modulated by the metastasis associated protein CAPL. AB - BACKGROUND: Several recent investigations have shown that the expression of the CAPL protein seems to be of importance in the metastatic potential in some types of cancer. However, the mechanisms behind this and other biological functions of CAPL are still largely unknown. The aim of the present work was to investigate whether CAPL could affect the expression of candidate proteolytic facilitators of the metastatic process, i.e. matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). MATERIALS AND METHODS: A highly metastatic osteosarcoma cell line with a high expression of CAPL was transfected with either a vector containing a ribozyme against this transcript, or with the vector alone as a control. The expression of MMPs and TIMPs was investigated with ELISA and gelatin zymography. RESULTS: The cell-line with a low CAPL expression (III-14) responded to bFGF treatment by an increased synthesis of MMP-1 and MMP-9 and to Il-1 alpha treatment by an increased synthesis of MMP-9. In contrast, the cell-line with a high CAPL expression (pH beta-1) did not respond with an altered expression of these MMPs. Neither of these two cell-lines responded with an altered expression of MMP-2. bFGF treatment resulted in an increased expression of TIMP-1 in both cell-lines, while Il-1 alpha treatment resulted in a decreased production of TIMP 1 in pH beta-1 cells, and III-14 cells were unaffected. CONCLUSIONS: The CAPL protein expressed in cell-cultures appear to block the MMP induction by bFGF and Il-1 alpha. However, the induction of TIMP-1 by bFGF must proceed through a pathway different from the MMP induced pathway, i.e. a pathway unaffected by CAPL. In addition, CAPL appeared to act in synergy with Il-1 alpha to reduce the synthesis of TIMP-1. PMID- 9858901 TI - Isolation of a variant which has reduced Src activity from SH-SY5Y neuroblastoma cells. AB - SY-HMs was obtained from SH-SY5Y neuroblastoma cell line transfected with an expression vector, pRc/CMV. SY-HMs showed much lower viability than the parent cells when cultured in low-serum medium. Drug sensitivity examined by MTT method revealed that this clone was highly sensitive to herbimycin A, a Src kinase inhibitor. Consistently with this, tyrosine phosphorylation of Src in this clone was profoundly reduced compared to that in the parent cells, suggesting that the activity of Src is seriously impaired in SY-HMs. It is thus possible that active Src is necessary for the survival of SH-SY5Y cells in low-serum medium. SY-HMs would be useful for further investigation of the role of Src in neuroblastoma cells. PMID- 9858902 TI - p53, Bax and Bcl-2 in vivo expression in the murine thymus after apoptogenic treatments. AB - BACKGROUND: Neoplasia can results from a lack of cell elimination by apoptosis. In order to determine if mechanisms controlling apoptosis are disturbed during neoplastic transformation in a model of murine radio-induced thymic lymphomas, we have assessed the kinetics of p53, Bax and Bcl-2 in situ expression after induction of thymic apoptosis by irradiation or glucocorticoids at first in normal mice. MATERIALS AND METHODS: TUNEL method was used for in situ detection of apoptosis and protein expression was determined by indirect immunohistochemistry. RESULTS: After hydrocortisone injection, levels of p53 and Bax, but not Bcl-2, expression were raised. A whole body sublethal irradiation led to an increase of p53 and Bcl-2, but not Bax, expression. CONCLUSIONS: This is the first in vivo report of in situ protein expression in the thymus after apoptogenic treatments of mice. The results suggest that Bax could be involved in glucocorticoid-mediated apoptosis. The increased levels of Bcl-2 expression are discussed. PMID- 9858903 TI - Fluorescent location of lung tumour cells. AB - Cells were collected on glass slides by touching tumour surfaces (A) and normal regions (B) of the lung. The slides were stained with a nuclear stain and a fluorescent probe for a tumour associated cell surface protein. The (B) slides from the normal regions lacked fluorescent epithelial cells. The tumour slides (A) contained typical tumour cells and dyskaryotic cells which exhibited cell surface fluorescence. PMID- 9858904 TI - HMGIC expression patterns in non-small lung cancer and surrounding tissue. AB - BACKGROUND: So far no powerful molecular markers have been found allowing an early detection of lung cancer enabling a higher rate of curative cancer therapy. Based on its expression patterns in other malignant tumors the expression of the high mobility group protein gene HMGIC is a possible new candidate for such a diagnostic marker. MATERIALS AND METHODS: The HMGIC expression patterns were determined in samples from 19 non-SCLC, 14 corresponding non-malignant adjacent lung, and 7 pleural tissues by hemi-nested RT-PCR followed by the detection of HMGIC specific amplification products by Southern blot hybridization. RESULTS: HMGIC expression was found in 14/19 non-SCLC samples. In contrast, expression was not detectable in any of the non-malignant adjacent lung or pleural tissues. CONCLUSIONS: These results suggest that HMGIC expression may be a potential new and powerful indicator for lung cancer. An HMGIC expression assay may be applicable to sputum or broncho-alveolar lavage fluid samples facilitating early detection of lung cancer. PMID- 9858905 TI - The effect of interferon and anti-CD44 antibody on mouse glioma invasiveness in vitro. AB - In this study the effect of interferon and anti-CD44 antibody on the invasiveness of mouse glioma G-26 cells was evaluated. We confirmed the glial nature of G-26 glioma cells (G-26) in vitro and in vivo using immunohistochemistry: G-26 stained strongly for S-100 and stained weakly for glial fibrillary acidic protein (GFAP). Immunohistochemical evaluation for CD44 adhesion molecule showed that G-26 was positive both in vitro and in vivo. Weakly positive punctate staining for CD44 was seen in the cytoplasm of all viable glioma cells and focally strong staining was observed in a membranous pattern in the invading glioma cells. Evaluation of untreated G-26 cells using an in vitro invasion assay showed that they were able to digest a Matrigel matrix and to invade through an 8 microns microporous membrane. Treatment of the G-26 glioma cells for 3-4 days with mouse interferon alpha/beta at 8 x 10(2) or 8 x 10(3) mu/ml resulted in a significant decrease of invasiveness: 68.8% (p < 0.05) and 32.8% (p < 0.001) of cells, respectively, remained invasive when compared to control. Treatment of G-26 with antibody to the CD44 adhesion molecule significantly decreased invasiveness with 39.4% (p < 0.001) of cells remaining invasive when compared to control. We feel that both of these approaches, each of which produced significant inhibition of G-26 glioma cell invasion should be further evaluated for their usefulness in antiglioma therapy. PMID- 9858906 TI - Differential paclitaxel-induced cytotoxicity in rodent and human hepatoma cell lines. AB - Hepatoma is the leading cause of death in male cancer patients in Taiwan. In this study, we examined the effect of Paclitaxel on the in vitro growth of 2 rodent and 4 human hepatoma cell lines. Differential Paclitaxel-induced cytotoxicity was observed among hepatoma cell lines. In Paclitaxel-sensitive Hep3B and N1S1 cells, Paclitaxel-induced cytotoxicity was dose- and time-dependent. The effective doses of Paclitaxel were in the range 0.1-1.0 microM. Flow cytometric analysis showed that Paclitaxel-treated hepatoma cells were arrested in G2-M phases prior to apoptosis. In addition, growth inhibition by Paclitaxel was accompanied by an increase in the expression of proliferating cell nuclear antigen (PCNA) in hepatoma cells. For Paclitaxel-resistant hepatoma cells, cytostatic response and/or polyploidization was observed. Our results indicated that two thirds of the hepatoma cell lines examined showed some degree of resistance to Paclitaxel treatment in vitro. The expression of p53 gene had no direct effect on Paclitaxel induced cytotoxicity. The expression of PCNA and the development of polyploidization appear to be good markers for measuring Paclitaxel response. These findings suggest that Paclitaxel alone appears to by cytostatic to hepatoma cells, combination of Paclitaxel with other chemotherapeutic agents may show better cytotoxic effects. PMID- 9858907 TI - Endomitotic index of megakaryocytes measured by flow cytometry helps to diagnose hematological disorders with abnormal platelet counts. AB - Platelet production is a regulated phenomenon. Indeed, megakaryocyte volume is inversely correlated to the platelet count not only in normal individuals and immune thrombocytopenic purpura patients, but also, and surprisingly, in chronic myeloid leukemia patients. Patients with polycythemia vera and essential thrombocythemia are located outside this regression line. Herein, we describe morphometrical data confirmed by the flow cytometric measurement of the megakaryocyte endomitotic index (EI). The EI is a value which reflects the mean ploidy of megakaryocytes and corresponds to the mean of (?log2 DNA content expressed in N?-1). In this study, the megakaryocyte endomitotic index of 14 normal individuals was compared to those of chronic myeloid leukemia (CML) patients (n = 16), immune thrombocytopenic purpura (ITP) patients (n = 11), essential thrombocythemia (ET) patients (n = 10) and polycythemia vera (PV) patients (n = 12). The megakaryocyte EI was significantly lower in CML patients than in normal individuals. In contrast, in ET, PV and ITP patients, megakaryocyte EI was higher than in normal individuals. An inverse relationship between the endomitotic index estimated by flow cytometry and the mean megakaryocyte volume performed by morphometry was observed in normal individuals, CML and ITP patients. In conclusion, the endomitotic index is higher in ITP, ET and PV patients and lower in CML patients when compared to normal individuals and is an interesting tool which can help to diagnose rapidly hematological disorders with abnormal platelet counts. PMID- 9858908 TI - Modulating the antitumor immunity of MBT-2 murine bladder tumor bearing mice by postoperative administration of interferon-alpha. AB - This study was conducted mainly to investigate the effect of interferon-alpha (IFN-alpha) on the antitumor immunity of a tumor bearing host (TBH) when postoperatively administrated with or without lethally irradiated autologous tumor cells. Using the C3H/He-MBT-2 murine bladder tumor model, a status of postoperative residual tumor was mimicked by rechallenging tumor cells 24 hours after resecting the day-17 tumor. Using immunohistochemical analysis we demonstrated that after treating with lethally irradiated MBT-2 tumor cells (IRMBT-2) + IL-2 cells of CD4+, CD8+, CD44+ and CD11b+ phenotypes prominently infiltrate the subcutaneous local injection sites. In contrast, only scanty immune responding cells could be seen locally if treated with IRMBT-2 + IFN-alpha 2b, albeit in the presence of interleukin-2 (IL-2). However, the spleens of D17TBM treated with IRMBT-2 + IFN-alpha 2b contained the highest percentage of CD44+ memory T cells and cells of the CD11b+ phenotype; moreover, their natural killer (NK), lymphokine activated killer (LAK) and cytotoxic T lymphocytes (CTL) activities were significantly augmented. The results of in vivo tumor rechallenge revealed that administration of IFN-alpha, either alone or combined with IRMBT-2, could both effectively suppress the outgrowth of perioperative rechallenged tumor cells as well as prolong the survival of TBH. We conclude that despite the presence of autologous tumor vaccine, postoperative administration of IFN-alpha can further enhance the antitumor immunity of TBH and therefore can be an effective adjuvant therapy to improve the therapeutic results of surgery on a tumor bearing host. PMID- 9858909 TI - Induction of apoptosis and inhibition of proliferation in human tumor cells treated with extracts of Uncaria tomentosa. AB - Growth inhibitory activities of novel water extracts of Uncaria tomentosa (C-Med 100) were examined in vitro using two human leukemic cell lines (K562 and HL60) and one human EBV-transformed B lymphoma cell line (Raji). The proliferative capacities of HL60 and Raji cells were strongly suppressed in the presence of the C-Med-100 while K562 was more resistant to the inhibition. Furthermore, the antiproliferative effect was confirmed using the clonogenic assay, which showed a very close correlation between C-Med-100 concentration and the surviving fraction. The suppressive effect of Uncaria tomentosa extracts on tumor cell growth appears to be mediated through induction of apoptosis which was demonstrated by characteristic morphological changes, internucleosomal DNA fragmentation after agarose gel electrophoresis and DNA fragmentation quantification. C-Med-100 induced a delayed type of apoptosis becoming most dose dependently prominent after 48 hours of exposure. Both DNA single and double strand breaks were increased 24 hours after C-Med-100 treatment, which suggested a well-established linkage between the DNA damage and apoptosis. The induction of DNA strand breaks coupled to apoptosis may explain the growth inhibition of the tumor cells by Uncaria tomentosa extracts. These results provide the first direct evidence for the antitumor properties of Uncaria tomentosa extracts to be via a mechanism of selective induction of apoptosis. PMID- 9858910 TI - Production and characterisation of a recombinant single-chain anti ErbB2-clavin immunotoxin. AB - We generated a recombinant immunotoxin, named scFv(MGR6)-Cla, composed of the Fv region of an anti ErbB2 monoclonal antibody (MGR6) fused to clavin, a type 1 ribosome-inactivating protein (RIP) from Aspergillus clavatus. ErbB2 is a tyrosine kinase receptor which is overexpressed in most adenocarcinomas; clavin is a 17 kDa ribonuclease which inhibits protein synthesis by inactivating ribosomes. A recombinant DNA construct containing the cDNA of the single chain Fv fragment (scFv) of the MGR6 antibody fused to the clavin cDNA, was expressed at high levels in Escherichia coli as an insoluble fusion protein containing an N terminal affinity tag of six consecutive histidine residues. Inclusion bodies were denatured and the recombinant fusion protein was purified under denaturing conditions by single-step purification using immobilised metal ion affinity chromatography (IMAC). The purified immunotoxin was renatured at high yield and histidine tag removed by digestion with enterokinase. The purity of the immunotoxin obtained after refolding was confirmed by SDS-PAGE, RP-HPLC, GPC-HPLC and N-terminal sequence analysis. Cell-free protein synthesis inhibition and binding assays showed that both clavin and scFv(MGR6) maintained their properties after refolding. PMID- 9858911 TI - Antitumor efficacy of nedaplatin, a novel platinum complex, with cyclophosphamide in murine and human tumor model. AB - BACKGROUND: The antitumor efficacy of the combination of Nedaplatin (NDP) with cyclophosphamide (CPM) was evaluated using murine and human carcinoma implanted mice. NDP was developed as a second generation platinum complex. Because of its superior antitumor activity and lower nephrotoxicity than Cisplatin (CDDP), we also compared the antitumor activity of NDP plus CPM with that of CDDP plus CPM. MATERIALS AND METHODS: Various doses of NDP or CDDP (1/4 to 1 maximum tolerated dose; MTD) and CPM (58 or 78 mg/kg) were injected once via the tail vein into mice implanted with Lewis murine lung carcinoma or Ma44 human lung carcinoma. RESULTS: Simultaneous administration of NDP or CDDP with CPM resulted in synergistically enhanced inhibition of tumor growth and prolonged survival in comparison with therapy using only NDP, CDDP or CPM. The combination therapy of NDP with CPM showed a superior survival effect with frequent long-term tumor-free survivors in comparison to that of CDDP plus CPM without increased hematotoxicity. The augmented antitumor efficacy of the combination of NDP with CPM was also demonstrated against Ma44 human lung carcinoma. CONCLUSION: The results suggested the effectiveness of using a combination of NDP with CPM for clinical therapy. PMID- 9858912 TI - 11 beta-hydroxysteroid dehydrogenase type II in human colon: a new marker of fetal development and differentiation in neoplasms. AB - The colon plays an important role in water and electrolyte homeostasis and is a major target tissue for aldosterone. 11 beta-Hydroxysteroid dehydrogenase type II enzyme (11 beta HSD2) confers specificity to the non-selective mineralocorticoid receptor by inactivating glucocorticoids, thus allowing binding by mineralocorticoids. Using immunohisto/cytochemistry and Northern blot analysis, we examined 11 beta-HSD2 expression in human fetal colon (23 cases ranging in age from 15 to 40 weeks gestation), neonatal colon (2 cases, 6 and 12 months old), normal adult colon (15), adenoma (35), adenocarcinoma (34) and the human colonic epithelial cell line T84, in the presence or absence of sodium butyrate, to study the correlation between enzyme expression and cellular or neoplastic differentiation. In fetal colon, weak 11 beta-HSD2 immunoreactivity was detectable in superficial epithelium from 25 weeks gestation, but normal adult levels were apparent only after 40 weeks gestation, suggesting that fully developed aldosterone induced electrolyte transport can occur only at late gestational stages. In adult normal colon, superficial absorptive cells at the top of crypts were strongly positive for 11 beta-HSD2, whereas immunoreactivity was weak in adenomas and carcinomas, and the pattern of localization varied among patients. Northern blot analysis performed on 4 cases of adenocarcinoma also demonstrated lower levels of mRNA than autologous non-neoplastic colonic mucosa. In carcinomas, 11 beta-HSD2 immunoreactivity was more frequently detected in differentiated structures, such as those forming glandular or tubular structures. Studies with T84 cells showed that expression of 11 beta-HSD2 was markedly enhanced with the addition of sodium butyrate, a well known inducer of cell differentiation in colonic epithelia. These results suggest that the expression of 11 beta-HSD2 is associated with differentiation or maturation in human colonic epithelia and that the enzyme may serve as a useful marker in development and disease. PMID- 9858913 TI - Gene-specific damage produced by topoisomerase inhibitors in human lung cancer cells and peripheral mononuclear cells as assayed by polymerase chain reaction stop assay. AB - We have previously reported that the polymerase chain reaction (PCR)-stop assay is an useful technique for investigating gene-specific damage induced by cisplatin, and that the degree of gene-specific damage sustained by peripheral blood mononuclear cells (MNC) when exposed to cisplatin in vitro predicts the response to cisplatin-based chemotherapy. In the current study, we investigated whether PCR-stop assay was suitable for investigating gene- specific damage induced by the topoisomerase I inhibitor CPT-11 or the topoisomerase II inhibitor VP-16, in the human lung cancer cell lines PC-7 and H69, respectively. The cells were incubated with CPT-11 or VP-16 for 24 hours in vitro and the hypoxanthine phosphoribosyl transferase gene was amplified by PCR. Amplification of a 2.7kb fragment of this gene was clearly inhibited by each drug in a dose dependent manner. The concentration which reduced amplification by 63% (D63, the dose that produces an average of one lesion per single strand of the 2.7kb fragment), was 318 micrograms/ml for PC-7 treated with CPT-11 and 35 micrograms/ml for H69 treated with VP-16. We also used PCR-stop assay to investigate gene-specific damage induced by CPT-11 or VP-16 in adenocarcinoma cells from pleural effusions and MNC from freshly isolated blood obtained from 4 patients with lung cancer. Between-patient variations in the extent of gene-specific damage were observed in both types of cells. These results suggest that PCR-stop assay is suitable for the analysis of interindividual variations in the extent of gene-specific damage induced by topoisomerase inhibitors in human cells. PMID- 9858914 TI - Second primary cancers after resection of lung adenocarcinoma with ras gene mutation. AB - BACKGROUND: Ras gene mutations are associated with a poor prognosis in patients with adenocarcinoma of the lung. However, the association between cases with ras gene mutation and an occurrence of second primary cancer is unknown. MATERIALS AND METHODS: We examined 89 adenocarcinoma of the lung obtained from patients treated by curative resection, and four second primary cancers, for mutation at codons 12, 13, and 61 of three ras oncogenes using polymerase chain reaction and oligonucleotide hybridization techniques. RESULTS: Ras gene mutations were detected in 13 cases. Six patients died from recurrent disease within 3 years. Of the seven patients who survived over 5 years, three patients demonstrated a metachronous second primary cancer after the operation for lung cancer, and one patient had synchronous laryngeal cancer. Two of the cases with second primary cancer demonstrated the ras gene mutation. One had the same mutation as that of the primary lung cancer, and the other had a different mutation from the first lung cancer. CONCLUSION: Ras gene mutations play an important role in tumor progression of lung adenocarcinoma and also might have an role in the carcinogenesis in respiratory tract cancer. PMID- 9858915 TI - Bystander effect caused by cytosine deaminase gene and 5-fluorocytosine in vitro is substantially mediated by generated 5-fluorouracil. AB - Because it appears impossible to transfer toxic genes to all the cells of a cancer, the bystander effect is critical to induce effective antitumor effects. In the present study, possible in vitro mechanisms of the bystander effect by the cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) were investigated. CD transduced cancer cells exhibited much higher sensitivity to 5-FC compared to parental cells. CD-transduced cells caused killing of neighboring parental cells in the presence of 5-FC, irrespective of direct cell-to-cell contact. Media conditioned by CD-transduced cells and 5-FC contained considerable amounts of 5 fluorouracil (5-FU) and exhibited profound cytotoxicity on parental cells. Furthermore, this killing ability of conditioned media correlated well with 5-FU levels converted from 5-FC by CD-transduced cells. CD was shown not to be secreted into media from cells. These results indicate that diffusible 5-FU plays the substantially causative role in the in vitro bystander effect caused by the CD/5-FC system. PMID- 9858916 TI - Potential coanthracyclinic activity of pyridylmethylene-2-indolinones. AB - The paper reports the cytotoxic activity of pyridylmethylene-2-indolinones previously described as cardiotonics and the synthesis of three analogs of the most potent cytotoxic agent. Some of these compounds could be useful, when associated with anthracyclines, to reduce the cardiotoxicity of these potent antitumor drugs. PMID- 9858917 TI - cAMP stimulates the bystander effect in suicide gene therapy of human choriocarcinoma. AB - In gene therapy, tumor cells expressing herpes simplex virus thymidine kinase (HSV-tk) are sensitive to ganciclovir (GCV) and HSV-tk positive cells exposed to GCV are lethal to adjacent HSV-tk negative cells. This phenomenon has been called the bystander effect, and the gap junction is thought to mediate it. In this study, sensitivity to GCV and bystander effect in a human choriocarcinoma cell line, BeWo, transfected with HSV-tk were investigated. Furthermore, the effect of 8-bromo-cAMP on bystander effect and connexin40 gene transcription were examined. HSV-tk positive cells were sensitive to GCV at the concentration of 10 micrograms/ml in a time-dependent manner. The growth of HSV-tk negative cells was inhibited when the population of cultured cells contained more than 10% HSV-tk positive cells and 8-bromo-cAMP enhanced bystander effect. 8-bromo- cAMP increased connexin40 mRNA expression and gap junctional intercellular communication. PMID- 9858918 TI - Adenovirus mediated thymidine kinase gene therapy may enhance sensitivity of ovarian cancer cells to chemotherapeutic agents. AB - The current treatment concept of ovarian cancer consists of radical surgery with subsequent chemotherapy. We have shown that adenovirus (ADV) mediated thymidine kinase (TK) gene transduction of cisplatin-resistant human ovarian cancer xenotransplanted into nude mice followed by ganciclovir (GCV) administration leads to prolongation of survival or cure. In this study the interaction of ADV TK gene therapy and selected chemotherapeutic agents commonly used for the treatment of ovarian cancer was investigated in three ovarian cancer cell lines with different growth patterns. Toxicity and cell killing efficacy of gene therapy, chemotherapy and their combinations with different concentrations and time intervals were measured by a 3-(4,5- dimethylthiazol)-2,5-diphenyl tetrazolium bromide (MTT) based assay. A slightly increased resistance to gene therapy was observed in cells pretreated with chemotherapy. Removal of the drugs restored the previous susceptibility of the cells to gene therapy. No antagonism was observed with gene therapy followed by chemotherapy. The concomitant application of gene therapy and chemotherapy resulted in a higher rate of cell death than the interval therapy. A dose dependent synergistic interaction was observed only for the combination of gene therapy and the topoisomerase 1 inhibitor topotecan. This synergistic effect was still seen even if the chemotherapeutic agent was added 72 hours later. Our data demonstrate that in addition to its own therapeutic efficacy, ADV-TK based gene therapy may enhance the effect of subsequent chemotherapy while up-front chemotherapy was disadvantageous. PMID- 9858919 TI - Preservation of metastatic ability of colorectal tumor cells stratified by inducibility of endogenous tumor necrosis factor after orthotopic transplantation in nude mice. AB - BACKGROUND: Inducibility of endogenous tumor necrosis factor (TNF) by colorectal tumor cells can be regarded as a novel prognostic factor in terms of distant metastasis or local recurrence following curative operation, especially at Duke's stage C. In this study, the metastatic ability of human colorectal tumor cells stratified by inducibility of endogenous TNF was analyzed by orthotopic transplantation in nude mouse. METHODS: Fifty three cases of freshly resected colorectal tumor specimens cut into about 50mg pieces were inoculated into the cecum wall of nude mice. Two to nine months after transplantation, tumor growth on this wall and metastases to the peritoneal wall as well as to the liver were assessed. RESULTS: Of forty one evaluable cases, successful transplantation was observed in twenty nine (70%), and metastases to the peritoneal wall or to the liver was found in thirteen (32%), or eight cases (20%), respectively. In the twenty nine cases with local tumor growth, incidence of the liver metastases in nude mice when tumor specimens from patients with liver metastases (4/8) were used was significantly higher than that from patients without liver metastases (4/21) (P = 0.096). Inducibility of endogenous TNF was separately analyzed in fourteen of the evaluable twenty nine cases. Seven cases belonged to the high group and seven to the low group in terms of the amount of endogenous TNF secreted by tumor cells. Incidence of metastases in mice was 1/7 in the high group and 6/7 in the low group, and there was a statistically significant difference between liver metastases in mice and inducibility of endogenous TNF by colorectal tumor cells (p = 0.0057). CONCLUSION: From these results, it is strongly suggested that inducibility of endogenous TNF by colorectal tumor cells can affect a patient's prognosis since it regulates metastatic ability to the liver. PMID- 9858920 TI - The amount and proteolytic content of vesicles shed by human cancer cell lines correlates with their in vitro invasiveness. AB - Cancer cells are known to shed extracellular membrane vesicles both in vitro and in vivo. To analyse their possible involvement in the metastatic behaviour of tumours, we measured the Matrigel invasion capability and amounts of vesicles shed by four human tumour cell lines (8701-BC, MCF-7, MDA-MB-231 and HT-1080), and by MCF-10A, an immortalised human breast cell line. The proteolytic activity content of vesicles was analysed by gelatin and casein zymographies. While MCF 10A cells do not release a measurable amount of vesicles, all tumour lines analysed, when cultured in presence of serum, shed vesicles rich in MMP-9. Other vesicle-associated proteinases include MMP-2 and uPA. Amounts and proteolytic activities of shed vesicles correlate with the in vitro invasiveness of cells. Since vesicles appear to promote the proteolytic cascade required for the localised degradation of the extracellular matrix, their shedding from cancer cells might represent an important feature of tumour progression. PMID- 9858921 TI - Synthesis and in vitro cytotoxicity of lipidic alcohols and amines. AB - General methods for the conversion of unsaturated fatty acids into alcohols and amines and the preparation of lipidic 1,2-diamines were developed. The in vitro cytotoxicity of the synthetic lipidic compounds was tested against two different cell lines (P388 and NSCLCN6). Oleyl amine was the most active among the lipidic alcohols and monoamines. However, the saturated lipidic 1,2-hexadecanediamine exhibited the highest cytotoxicity (IC50 0.1 microgram/ml and 1.1 micrograms/ml). PMID- 9858922 TI - Submicromolar paclitaxel induces apoptosis in human gastric cancer cells at early G1 phase. AB - Paclitaxel induced apoptosis has been reported in many cancer cell lines, but the relationship between G2/M phase arrest and apoptosis is not clear. We have reported that low dose (10 nM) paclitaxel induced apoptosis in gastric cancer cells without G2/M phase arrest. In this study, SC-M1 gastric cancer cells were synchronized at early G1, late G1, S and G2/M phases by staurosporine, mimosine, hydroxyurea and berberine, respectively. We have found that paclitaxel could not induce apoptosis of gastric cancer cells in late G1, S and G2/M phases; however, paclitaxel induced apoptosis in the early G1 phase. Our results suggest that cells arrested at G2/M phase by paclitaxel eventually entered the early G1 phase then proceeded to apoptosis. PMID- 9858923 TI - Scanning electron microscopy of aberrant crypt foci in human colorectal mucosa. AB - BACKGROUND: Aberrant crypt foci (ACF) are clusters of morphologically altered crypts which can be observed by light or stereomicroscopy on the mucosal surface of the colon after staining with methylene-blue. They probably represent one of the earliest events in human colorectal carcinogenesis. The main purpose of the present study was to observe the surface features of aberrant and normal colonic crypts in humans using scanning electron microscopy (SEM) in order to find and measure differences between aberrant and normal. MATERIALS AND METHODS: Fifteen mucosal specimens containing ACF and 8 with normal mucosa taken from patients operated on for colon cancer were observed under a scanning electron microscope. RESULTS: By SEM ACF were easily observed on the mucosal surface, because they showed a well defined border and were elevated on the mucosal surface. Under higher magnification luminal openings of aberrant crypts had a larger overall average diameter than normal (37.6 microns +/- 13.5, mean +/- SD, vs 15.9 microns +/- 4.9, P = 0.001), though when crypt multiplicity of ACF (number of crypts per ACF) was higher, the diameter of luminal openings tended to be smaller and similar to those of normal crypts, with weak negative correlation between crypt multiplicity of ACF and mean diameter of aberrant luminal openings (r = 0.27). Finally, the mucosal surface among aberrant crypts was flattened because of a loss of microvilli. in conclusion, scanning electron microscopy allows a better definition of the topological features of aberrant crypt foci than light or stereomicroscopy. PMID- 9858924 TI - Apoptosis in different stages of human oogenesis. AB - Apoptosis is an active form of cell death characterized by a series of morphological changes that become particularly evident at the ultrastructural level. The majority of ovarian germ cells undergo degeneration during prenatal and reproductive life and only in recent studies has it been demonstred that this drop is due to an apoptotic process. We evaluated this process during human oogenesis in prenatal life and we studied the ultrastructural changes that occur in apoptosis in various phases of the meiotic process. From our observations it is clear that apoptosis involves two main phases of the meiotic process: an earlier one concerning the oogonia and oocytes in the preleptotene stage, and a later one that mainly concerns the oocytes in the pachytene stage. PMID- 9858925 TI - Does AK-2123 (Senazole) have sensitizing effects on radiation, cisplatin and hyperthermia under aerobic conditions in vitro? AB - We investigated the sensitizing effects of AK-2123 (Senazole) on the interaction of radiation, cisplatin and hyperthermia under aerobic conditions in the rat yolk sac cell line NMT-1R in vitro. The effects were assessed by clonogenic assay. A cytotoxic effect of AK-2123 after 24 hours exposure was observed as a function of the dose. For NMT-1R cells, the ID70 of AK-2123 was 400 micrograms/ml for 24 hours exposure, which was employed for subsequent combined treatments. Although a statistically significant increase in the G1 cell fraction was observed after AK 2123 treatment with a dose of ID70 (p = 0.02) no enhancing effect of AK-2123 on radiation, cisplatin or heat response curves was detected under aerobic conditions in vitro. PMID- 9858926 TI - Apoptosis during the development of radiogenic thymic lymphomas: effects of treatments inhibiting lymphoma development. AB - INTRODUCTION: Whole body fractionated irradiation induces thymic lymphomas in C57BL/Ka mice after a latent period during which preleukemic cells progressively transform into leukemic cells within an abnormal thymic microenvironment. A bone marrow graft or repeated cytokine injections prevent lymphoma development. We think that these treatments restore altered mechanisms controlling apoptosis. MATERIALS AND METHODS: Apoptosis was analyzed by flow cytometry in thymocytes from different groups of mice (control, preleukemic, prevented mice). RESULTS: The apoptotic rates did not change in freshly isolated thymocytes from different experimental groups. However, after culture, the level of apoptosis increased in preleukemic thymuses; and returned to normal value in cultured thymocytes from irradiated mice after lymphoma preventing treatments. Furthermore, thymic microenvironmental factors can control thymocyte apoptosis. CONCLUSION: We propose that after leukemogenic irradiation, there is an increase of cells with an activated suicide program, but that alterations of thymic environmental factors rescue them from apoptosis, allowing their further neoplastic transformation. PMID- 9858927 TI - Inhibition by Porphyromonas gingivalis LPS of apoptosis induction in human peripheral blood polymorphonuclear leukocytes. AB - Many factors, such as cytokines and bacterial products, are known to affect the life-span of polymorphonuclear leukocytes (PMNs). We investigated here whether Porphyromonas gingivalis (P.gingivalis) LPS slows down the apoptotic process of PMNs. During incubation in regular culture medium, the viability of PMNs progressively declined, producing apoptotic cells, characterized by chromatin condensation, internucleosomal DNA fragmentation, cell shrinkage and blebbing of plasma membrane. P.gingivalis LPS significantly increased the viability of PMNs and reduced the production of fragmented DNA, as efficiently as E.coli LPS. The present study suggests that the retardation of PMNs apoptosis by P.gingivalis LPS may modulate the restoration of acute inflammation in the periodontal tissues, where activated PMNs accumulate. PMID- 9858928 TI - Effects of vitamin C on arylamine N-acetyltransferase activity in human liver tumor cells. AB - Arylamine N-acetyltransferase (NAT) activity in a human liver tumor (heptoma) cell line was inhibited by vitamin C. Using high performance liquid chromatography, NAT activity on the acetylation of 2-aminofluorene and p aminobenzoic acid was examined. Two assay systems were performed, one with cellular cytosols, the other with intact liver tumor cell suspensions. The NAT activity in a human liver tumor cell line was inhibited by vitamin C in a dose dependent manner in both types of examined system: i.e. the greater the concentration of vitamin C in the reaction, the greater the inhibition of NAT activities in both systems examined. The data also indicated that vitamin C decreased the apparent Km and Vmax of NAT enzymes from human liver tumor cells in both systems examined. This report is the first demonstration which showed vitamin C effect on human liver tumor cell NAT activity. PMID- 9858929 TI - Radical intensity and cytotoxic activity of curcumin and gallic acid. AB - Natural phenolic compounds, curcumin and gallic acid, were compared for their cytotoxic activity in relation to their radical modulating activity. These two compounds induced apoptotic cell death in human promyelocytic leukemic HL-60 cells and human oral squamous carcinoma HSC-4 cells. Curcumin was more cytotoxic than gallic acid. Catalase reduced significantly the cytotoxic activity of gallic acid, but not that of curcumin. ESR spectroscopy demonstrated that curcumin produced radicals under alkaline conditions, scavenged the superoxide anion radical, and enhanced the radical intensity of sodium ascorbate at higher concentrations. As compared with curcumin, gallic acid produced higher amounts of radicals and more efficiently scavenged the superoxide anion radical. Gallic acid reduced the radical intensity of sodium ascorbate, suggesting a possible interaction between these two compounds. These data suggest that curcumin and gallic acid induce apoptosis by different mechanisms. PMID- 9858930 TI - Interaction of chlorpromazine with 2'-deoxyguanosine-5'-monophosphate by PM3 calculation. AB - Using the PM3 method, the interactions between chlorpromazine (CPZ) with 2' deoxyguanosine-5'-monophosphate (dGMP) were examined. We obtained the optimized geometrical structure of each CPZ, dGMP and a CPZ-dGMP system in both aqueous phase and gaseous-phase and investigated their geometric and electric changes and Force calculation. By Force calculation, three vibrations at 835, 800 and 737 cm 1 were assigned to the antisymmetric and symmetric P-O stretching vibration of PO3(2-) group in Z-dGMP, respectively. The vibrations at 889, 803 and 799 cm-1 due to the C5'-O-PO3(2-) stretching vibration shifted to their corresponding higher wave numbers, comparing to those of Z-dGMP alone. PMID- 9858931 TI - Topoisomerase-I inhibitor SN-38 can induce DNA damage and chromosomal aberrations independent from DNA synthesis. AB - BACKGROUND: SN-38 is the active metabolite of the topoisomerase-I (topo-I) inhibitor Irinotecan (CPT-11). Generally, topo-I inhibitors stabilize the complex between topo-I and DNA which collide with moving DNA replication forks, eventually leading to double stranded DNA damage. Therefore, topo-I inhibitors are regarded as S-phase specific. The present study investigated S-phase dependent and independent effects of SN-38. MATERIALS AND METHODS: Effects of exposure of A2780 cells to SN-38 (2 hours) were studied by assessing DNA/protein crosslinks, DNA damage and cytogenetic aberrations. RESULTS: A close correlation (r2 = 0.97) was established between drug-induced DNA/protein crosslinks and double stranded DNA breaks. Cytogenetic analysis revealed near maximum clastogenic effects already evident immediately following 2 hours drug exposure. However, qualitatively, chromatid breaks at 24 hours were different from those at 0 hours, in that at 24 hours they were associated with radial chromosome configurations and sister chromatid exchanges. CONCLUSION: The data corroborate that the S-phase dependent mechanism of action of topo-I inhibitors is also applicable to SN-38. The cytogenetic data indicate two distinct interactions of SN-38 with DNA: immediate induction of chromatid breaks independent from DNA synthesis, and induction of chromatid breaks associated with radial chromosome configurations dependent on DNA synthesis. PMID- 9858932 TI - Evidence that phosphatidylinositol 3-kinase and p70S6K protein are involved in differentiation of HL-60 cells induced by calcitriol. AB - 1,25-dihydroxyvitamin D3 (calcitriol) is not only an antirachitic agent, but also a well known regulator of cell differentiation. HL-60 promyelocytic leukemia cells differentiate to monocytes upon treatment with calcitriol. We describe here, that PI3-K inhibitors are able to block the differentiation induced by calcitriol in HL-60 cells. Also the downstream effector of PI3-K, p70S6K ribosomal protein kinase seems to be involved in HL-60 cell differentiation. PKC alpha and PKC delta are activated and translocated to the nucleus upon exposure of cells to calcitriol. However in our experiments the inhibition of PKC did not result in an inhibition of calcitriol induced differentiation of HL-60 cells. On the contrary, the use of thapsigargin, caused the differentiation process to stop. PMID- 9858933 TI - Morphological aspects of altered basement membrane metabolism in invasive carcinomas of the breast and the larynx. AB - In the present study we compared the localization of major basement membrane (BM) components and their mRNAs between invasive carcinomas of the breast (adenocarcinomas) and larynx carcinomas (squamous cell carcinomas, SCC), in order to determine the extent of BM production and deposition in malignant tumors of biologically different behaviour. Thus, breast carcinomas usually show a rapid locoregional/systemic spread, while the laryngeal SCCs normally show a more locally restricted growth pattern. While normal mammary glands and laryngeal mucosa revealed an intact epithelial BM as evidenced by a continuous linear staining for collagen IV, laminin-1, heparan sulfate proteoglycan (perlecan) and fibronectin-as well as collagen VII in the larynx mucosa-, this continuous staining was lost in the invasive carcinomas, however, affecting the two tumor types differently. In the breast carcinomas, a complete loss was seen even in well differentiated tumors affecting the various BM components similarly, while in the SCCs well differentiated carcinomas had retained significantly more BM material than poorly differentiated ones. In the SCCs, an "early" loss of collagen VII contrasted with a "later" loss of collagen IV, laminin, perlecan and fibronectin the extent of which was, however, associated with a decreasing degree of differentiation. In contrast to the protein findings, by use of the in-situ hybridization we observed a significant expression of mRNA for collagen IV, perlecan and fibronectin. The resulting pattern was comparable between both tumor types and not significantly related to the tumor cell differentiation. Both tumor cells and stroma cells were positively labelled with a more extensive labelling of the stroma cells. Our observations indicate a similar upregulation of the mRNAs for BM-components in breast and larynx carcinomas, but significant differences in the BM-protein deposition so that either major differences in presumed BM-proteolysis or further translational defects are suggested. Furthermore, it can be speculated that the far lesser amount of BM-material in the breast carcinomas may be linked to the more aggressive metastatic spread of those tumors, particularly when compared to the SCCs. PMID- 9858934 TI - Effects of dietary fiber on the rat intestinal mucosa exposed to low doses of a carcinogen. AB - BACKGROUND: Changes in morphological and immunohistochemical parameters were studied in the rat intestinal mucosa exposed to low doses of a carcinogen and administered with dietary fibers. METHODS: Tumors were induced by five subcutaneous injections of 1,2-dimethylhydrazine, 10 mg/kg rat, once a week. Rats were fed a semi-synthetic fiber-free diet (control) or a high-fiber diets (15%) derived from cellulose, tomato peels or white grape. The rats were sacrificed 24 weeks after the first carcinogen's injection. The ileum, colon and tumors were removed for the study. Areas of the mucosal stroma and of lymph infiltrations, and mitotic index were studied along with morphological parameters. Immunohistochemical parameters included determination of Ki-67 proliferating protein and apoptotic index. RESULTS: Areas of the stroma in colon tumors increased in rats fed tomato peels. Changes in areas of lymphoid infiltrates were related to the type of diet and tumor presence. Lymphoid infiltrations were found to be highly developed in the colon area close to tumors, especially in rats fed the white-grape diet. Mitotic index and Ki-67 protein increased significantly in the colon area close to a tumor and in tumors themselves without any relation to the fiber varieties consumed. Changes in the rate of apoptosis were not related to the preventive effect of diets: apoptotic index was high in tumors obtained from rats fed the high-cellulose diet with high tumor-preventive effects and also from rats fed the high-tomato-peel diet with low tumor-preventive effects. CONCLUSIONS: No morphological changes were found in the ileum of rats exposed to a carcinogen and fed different dietary fibers. In the colon, a carcinogen even in low concentrations inhibited the lymphoid system in the mucosa located far from the tumor or close to the tumor. An increase in the proliferation rate in the colon close to the tumor may reflect the development of precanceromatous processes or may be related to the effect of growth factors expressed by tumor cells. Finding adenoma-like dysplasia near tumors may be possible in early stages of the development of new tumors. In addition, activation of the lymphoid system of the colon following consumption to specific dietary fiber may be a mechanism by which fiber protect against cancer. PMID- 9858935 TI - The p53 and nm23-H1 genes are not deleted in oral benign epithelial lesions. AB - In an effort to identify genetic changes that may be the early hallmarks of epithelial cell overproliferation, we searched for p53 and nm23-H1 allelic deletions in oral benign epithelial lesions. In the study group were 25 benign epithelial lesions (lichen planus--17; leukoplakia--8; recurrent aphthous ulcers- 2; one specimen diagnosed as benign migratory glossitis). Among 21 samples analysed for exon 4 (p53 gene) LOH, only 6 were informative, with no loss of either allele. OF 23 samples tested for LOH at intron 6 of p53 gene, 8 were informative, again with no presence of LOH. For nm23-H1 gene, the analysis was performed on a total of 24 cases. Of them, 16 were informative, however, none exhibited LOH at this locus. In conclusion, whereas the presence of gross gene alterations (LOH) would have been definitive evidence for the involvement of p53 and/or nm23 in the hyperproliferation process, the absence of LOH does not exclude the presence of either smaller mutations, altered regulation of normal gene, or dysfunction at the level of wild type protein. Alternatively, p53 and nm23-H1 may have no relation to oral lesion formation, and cannot presently be considered as an early step in benign, tissue transformation. PMID- 9858936 TI - The influence of fatty acids on normal and transformed human liver cells in culture. AB - Mammals are incapable of the synthesis of certain polyunsaturated fatty acids and must fulfil their needs from plant sources. These polyunsaturates are further desaturated and chain elongated by the mammal, and quantitatively the most important site is the liver. The enzymes concerned with this process are the desaturases and these have been shown to be lost with cell transformation. We incubated normal human liver cells and a hepatoma cell line with fatty acids of differing degrees of unsaturation and chain length, to ascertain whether the transformed cells would exhibit the same response to the fatty acids as the normal liver cells. There were no differences in the way the two cell types reacted to the saturated fatty acid, but a mixed response to the monounsaturate. The two plant-derived polyunsaturates caused more of the hepatoma cells to die, and this effect was more pronounced with the longer chain more unsaturated compounds. The three fatty acids that are precursors for eicosanoid synthesis showed the least effect on the normal cells, but still caused hepatoma cell death to a degree very similar to that caused by the plant polyunsaturates. PMID- 9858937 TI - Transcriptional regulation of the alpha 4 integrin subunit gene in the metastatic spread of uveal melanoma. AB - Recently, expression of the alpha 4 integrin subunit has been shown to be inversely correlated with the invasive potential of B16 mouse epidermal melanoma. The purpose of this study was to establish whether expression of the human alpha 4 integrin subunit gene might be similarly regulated in human uveal melanoma which has varying degrees of invasiveness, and whether such modifications are determined by alterations in the transcriptional activity directed by the alpha 4 gene promoter. Two metastatic variants (MH5 and MH10) derived from a human uveal melanoma (SP6.5) were used. Expression studies were performed by transiently transfecting each of these cell lines with recombinant plasmids bearing various lengths of the alpha 4 promoter fused to the CAT reporter gene, and were further validated by Northern blot analyses of the alpha 4 transcript. Both transient transfection and mRNA analyses provided evidence that the transcriptional activity directed by the alpha 4 promoter sequences extending up to position -76 and -120 was indeed inversely correlated to the potential of uveal melanoma to yield metastasis. Experiments in electrophoretic mobility shift assay (EMSA) demonstrated that binding of the nuclear proteins that likely account for transcription of the alpha 4 gene to alpha 4.1 (namely Bp1, Bp2, Bp4, and Bp5) was dramatically reduced in uveal melanoma, but not in normal uveal melanocytes. These results highlight the fundamental function the alpha 4 integrin subunit may play in the ability of tumor cells to evade the primary tumor and form metastasis. PMID- 9858938 TI - Indomethacin inhibits kidney metastasis in bomirski melanoma-bearing hamsters, and modulates natural killer cytotoxic activity of tumor hosts in vivo and in vitro. AB - Natural killer (NK) cells are thought to play an important role in the control of metastatic dissemination. Therefore, stimulation of cytotoxic activity of NK cells against neoplastic cells could be preventive for metastatic spread. Bomirski amelanotic (Ab) melanoma of Syrian hamster is a transplantable tumor metastasizing preferably to the kidneys. During growth of the melanoma a significant depression of cytotoxic activity of NK cells of tumor hosts is observed. Treatment of melanoma-bearing hamsters with indomethacin provided in drinking water resulted in the increase of NK cytotoxic activity of blood cells and in the lower occurrence of kidney metastasis. Spleen cells obtained from healthy and melanoma-bearing hamsters were cultured in vitro with agents influencing NK activity. We found an augmentative effect of human interleukin 2 (IL2) and human tumor necrosis factor (TNF). We also observed the synergistic effect of IL2/TNF combination, which was present in both groups of animals. The stimulatory effects of cytokines could be potentiated by the additional supplementation of cultures with indomethacin. Similar experiments were performed on spleen cells isolated from the healthy and tumor-bearing animals treated in vivo with indomethacin. Also, in this group of hamsters in vitro stimulation of NK cell activity by the cytokines was effective. The studies presented may give insight into the pathogenesis of immune abnormalities seen in advanced stages of progression of Ab melanoma, and can provide an experimental basis for immunomodulation in this tumor model of spontaneous metastasis. PMID- 9858939 TI - Comparison of the effects of bischolylglycinatechloro-platinum(II) versus cisplatin on liver regeneration after partial hepatectomy. AB - BACKGROUND: Regional chemotherapy by intraportal administration has been envisaged as a useful strategy to prevent the high rate of recurrence after surgical removal of single liver tumors, even though it may inhibit liver regeneration. New cytostatic drugs, such as Bamet-H2-Na[Pt(cholylglycinate-O,N) (cholylglycinate-O) Cl]- have been developed to enhance the liver organotropism of antiproliferative agents. The aim of the present study was to compare the effects of Bamet-H2 and cisplatin on liver regeneration. MATERIALS AND METHODS: The ability of both drugs to inhibit liver cell proliferation was investigated using rat hepatocytes in primary culture. Two-thirds partially hepatectomized mice were used to measure the effects of glycocholic acid, cisplatin and Bamet-H2 on DNA synthesis by the regenerating liver in vivo. RESULTS: Up to 300 microM glycocholic acid, no effect on the growth of rat hepatocytes in primary culture was observed. By contrast, similar dose-dependent in vitro cytostatic effects of cisplatin and Bamet-H2 were found. No effect on mouse liver regeneration was found in animals receiving glycocholic acid. By contrast, both cisplatin and Bamet-H2 were found to induce a significant inhibition of DNA synthesis and slower recovery of liver mass. The effect of cisplatin was significantly stronger than that induced by Bamet-H2. Differences between these two compounds were also observed regarding the content of the drug in several tissues at short- and mid term. Both drugs were highly concentrated in the liver and kidney, with minor distribution in other tissues at 8 hours after the last injection of the drug. However, Bamet-H2 was more efficiently eliminated from the body in 5 days. Moreover, higher toxicity and lower survival were observed in the group of animals treated with cisplatin as compared to Bamet-H2. CONCLUSIONS: Our results indicate that in agreement with the previously reported effects on several tumor cells lines, cisplatin and Bamet-H2 are similarly efficient as cytostatic drugs in liver cells when these are continuously exposed to the compounds in vitro. However, in the in vivo situation Bamet-H2 is better tolerated by the animals and induces less inhibition of liver regeneration than cisplatin. This is probably due to the enhanced biliary elimination of Bamet-H2. PMID- 9858940 TI - Interaction between sodium 5,6-benzylidene-L-ascorbate and dopamine. AB - The interaction between sodium 5,6-benzylidene-L-ascorbate (SBA) and dopamine was investigated by three different parameters: radical intensity, prooxidant action and cytotoxic activity. Under alkaline conditions, SBA and dopamine produced doublet and quartet ESR signals, respectively. The addition of increasing concentrations of SBA completely scavenged the dopamine radical and replaced the latter with its own radical. On the other hand, dopamine accelerated the decay of SBA radical. These two compounds stimulated the methionine oxidation in culture medium, but in combination, their stimulation activities were weakened. Both of these two compounds dose-dependently reduced the viable cell number of human promyelocytic leukemics HL-60 cells. When these two compounds were mixed together before adding to HL-60 cells, both of their cytotoxic activities were diminished. The present study demonstrates the interaction between SBA and dopamine, which might modify their biological activities. PMID- 9858941 TI - High doses of thymosin alpha 1 enhance the anti-tumor efficacy of combination chemo-immunotherapy for murine B16 melanoma. AB - BACKGROUND: We have reported previously that combined chemo-immunotherapy with cyclophosphamide (CY), thymosin alpha 1 (T alpha 1) and low dose interferon alpha,beta (IFN alpha beta) has significant anti-tumour effects. Here, using mouse B16 melanoma as a model, we tested whether increasing the dose of T alpha 1 could increase the anti-tumour activity of triple combination chemo immunotherapy. MATERIALS AND METHODS: C57BL/6 mice were challenged subcutaneously with B16 melanoma cells and injected intraperitoneally with saline, CY (200 mg/kg, day 7), or CY with T alpha 1 (200, 600 or 6000 micrograms/kg/day, days 10 13) and IFN alpha beta (30,000 I.U., day 13). RESULTS: Chemo-immunotherapy with high dose (HD)-T alpha 1 caused complete tumour regression for 27.5 days after tumour cell injection (3.9 times longer than untreated controls) and delayed tumour relapse compared to all other groups. Moreover, it significantly increased the median survival time of treated mice, and cured an average of 23% of animals, while none was cured in any other group. Splenocytes from HD-T alpha 1-treated mice showed markedly increased cytotoxic activities against both YAC-l and autologous B16 tumour cells. HD-T alpha 1 treatment reversed the tumour-induced reduction in percentages of CD3 and CD4-positive splenocytes to non-tumour levels, and it increased percentages of CD8, B220 and IL-2R beta-positive cells to well beyond non-tumour controls. CONCLUSIONS: High doses of T alpha 1 improve anti-tumour efficacy of tnple chemo-immunotherapy against B16 melanoma. These effects appear to be mediated by stimulation of the host immune response. PMID- 9858942 TI - Resistance to apoptosis and cyclooxygenase-2 expression in a human adenocarcinoma cell line HT29 CL.19A. AB - BACKGROUND: COX-2 expression increases the tumorigenic potential of enterocytes. Tumorigenic effect is partially linked to an inhibition of programmed cell death which is one of the most important components in maintaining intestinal epithelium integrity. MATERIALS AND METHODS: We analyzed apoptosis in HT29 cl.19A cells cultured over 3 weeks, in the presence (10%) or in the absence of fetal bovine serum (FBS), by analysis of genomic DNA fragmentation after agarose gel electrophoresis, morphological measurement of apoptosis using DAPI chromatin staining, and transmission electron microscopy (TEM) to identify apoptotic cellular morphological changes. RESULTS: Regardless of the methods used, no apoptotic signs were observed for each culture condition, even if cells were cultured 3 weeks in the absence of FBS. CONCLUSION: Using HT29 cl.19A cells (untransfected cells), we found that intrinsic or constitutive COX-2 expression in adenocarcinoma cell line was associated with spontaneous resistance to apoptosis. PMID- 9858943 TI - Valproic acid inhibits proliferation and changes expression of CD44 and CD56 of malignant glioma cells in vitro. AB - Recently we were able to show that valproic acid (VPA) induces growth-arrest and differentiation of human neuroblastoma cells. Hence we investigated in vitro the antitumoral effects of VPA on malignant gliomas by determining cell proliferation and expression of CD56 and CD44 of human T98G, A172, 85HG66, 86HG39 and rat C6 cell lines. VPA at concentrations ranging from 0.1 to 1 mM strongly inhibited proliferation of A172, 86HG39, 85HG66 and C6 cells in a dose-dependent manner, whereas T98G cell growth remained unchanged. All human glioma cells were highly positive for CD44, whereas CD56 was differently expressed. After 7 days of incubation with 1mM VPA CD56 expression was markedly increased in T98G, A172 and 85HG66 cells, whereas CD44 expression was decreased in all human cell lines. These data suggest that VPA has antitumoral effects on malignant glioma cells. Therefore we consider VPA as a potent therapeutic agent for treatment of these tumors. PMID- 9858945 TI - 2'-2'-difluorodeoxycytidine: in vitro effects on cell-mediated immune response. AB - Little or no data are available on the immunotoxicity of the new deoxycytidine analogue, gemcitabine (2'-2'-Difluorodeoxycytidine, dFdC). The drug was tested on natural killer (NK), interleukin 2-activated killer (LAK) and antigen-dependent cytotoxic effector cells (CTL) activity. NK cells were treated for 16 hours and then tested against K562 cell line. LAK cells were pretreated for 16 hours before or after IL2 stimulation, and tested against DAUDI cells on day 4. CTL were pretreated for 16 hours on day-1 or on day 4 of coculture, and then tested against MT-2 on day 5. Cytotoxic activity was evaluated by a 4 hours 51Cr-release assay. The results indicate that dFdC inhibits markedly LAK or CTL generation, but does so less efficiently on "mature" LAK or CTL lymphocyte function and only slightly on NK cell activity. Therefore dFdC can be considered immunotoxic for either natural or antigen-dependent cell-mediated immunity. PMID- 9858944 TI - Effect of 4-hydroxynonenal, a product of lipid peroxidation, on NK susceptibility of human K562 target cells. AB - 4-Hydroxynonenal (HNE) is one of the major breakdown products generated by lipid peroxidation of cellular membranes. The level of lipid peroxidation and the concentration of its products are inversely related to the rate of cell proliferation and directly related to the level of cell differentiation. It has been reported that HNE inhibits DNA synthesis, ornithine decarboxylase (ODC) activity and c-myc expression in different leukemic cells lines. It has also been demonstrated that HNE inhibits proliferation and induces differentiation in HL60 cell line. In the present study the effects of HNE, at concentrations close to those found in the normal tissues, on the NK susceptibility of human K562 target cells were analyzed. Repeated treatments at 45 minutes intervals with 1 microM HNE were performed to maintain the cells in the presence of the aldehyde for 12 hours. The effect of HNE was compared with that obtained in Haemin-treated cells. HNE causes a strong inhibition of cells growth (53% vs. 34% with Haemin) without affecting cell viability. We further investigated the NK susceptibility of K562 cell line upon in vitro treatment with HNE. Cytotoxic activity of mononuclear cells (MNC) from peripheral blood of healthy donors was determined by 4 hours Cr51-release assay. The results obtained, expressed in terms of percentage of specific lysis at different E:T ratios and in terms of KC (10(6)) at the E:T ratio of 50:1, show that HNE treatment of K562 cells leads to a marked reduction of susceptibility to NK cells; this decrease is very close to that found in the K562 cells treated with Haemin used as inducer. Similar results were obtained using MNC pre-treated with beta-interferon (IFN) as effector cells. MNC show a reduced capacity to lyse HNE-treated cells also under the enhancing cytolytic effect of IFN. These results are in line with data obtained with several common inducers of differentiation such as DMSO, retinoic acid or others. PMID- 9858946 TI - A high-fat diet does not influence the growth of the Dunning R3327-H transplantable prostate adenocarcinoma. AB - BACKGROUND: The etiology of prostate cancer is currently a mystery. Several epidemiological studies suggest a link between dietary fat and prostate cancer. In vitro and in vivo studies support this evidence. Using the Dunning model of rat prostate cancer we hypothesized that a high-fat diet (20%) would increase the growth of the R3327-H tumor. MATERIALS AND METHODS: R3327-H tumors were implanted subcutaneously into male Copenhagen rats which were fed diets with 5 or 20% total fat. Tumors were allowed to grow for 16 weeks; they were then excised and weighed. The initial and final weights of the rats were also recorded. RESULTS: Statistical analysis revealed the level of dietary fat was a positive predictor of weight gain (p < 0.01). No effect on tumor growth was seen when compared to dietary fat, fiber type, or the interaction of fat and fiber. DISCUSSION: Growth of the R3327-H tumor, when implanted subcutaneously, is not affected by the level of dietary fat. PMID- 9858947 TI - Beneficial effects of a vanadium complex with cysteine, administered at low doses on benzo(alpha)pyrene-induced leiomyosarcomas in Wistar rats. AB - BACKGROUND: Vanadium is a potent environmental and body metal, possessing remarkable antitumor and antidiabetic properties. Vanadium salts and complexes have been widely investigated for their anticarcinogenic properties in experimental carcinogenesis. In the present study the antitumor effects of a new vanadium complex with cysteine in relation to identical doses of vanadyl sulfate and cysteine, in tumor bearing rats are investigated. MATERIALS AND METHODS: Male wistar rats were injected with benzo(alpha)pyrene and divided into four groups of 21 rats each. Control group was treated only with BaP. The first group(TR-1) was treated by vanadyl sulfate per os at daily doses of 0.5 mg of V/kg b.w per day. The second (TR-2) by cysteine at doses of 4.5 mg/kg b.w per day and the third group (TR-3), by the complex V(III)-cysteine at daily doses of V 0.5 mg/kg b.w (containing cysteine at concentrations of 4.5 mg/b.w). Treatment was started when tumors were developed (evidenced from a palbable mass at the site of Bap injection) and went on till death. Toxicological tests were performed in 27 rats divided into a control group and two test groups; T-1 administered with vanadyl sulfate at daily doses of 18.5 mg V/kg b.w and T-2 group with V(III)-cysteine complex at daily doses of 18.5 V/kg b.w, for 9 weeks. Mean survival time, death rate, tumor growth rate, the carcinogenic potency of BaP, and the anticarcinogenic potency in relation to histological findings in each treatment group were calculated in each group in order to evaluate the antitumor effects of the substances used. RESULTS: Vanadyl sulfate, cysteine and V(III)-cysteine exerted antitumor effects on leiomyosarcoma bearing Wistar rats. However, V(III) complex exerted much more potent effects than the other treatments, significantly prolonging mean survival time, retarding tumor growth rate and decreasing the carcinogenic potency of BaP in the TR-3 group, in comparison to the control and the TR-1 and TR-2 groups. Moreover V(III)-cysteine complex resulted in complete remission of 4 (19.7%) of the tumor bearing rats. Blood, urine, biochemical routine tests as well as autopsy did not reveal any toxic effects either of vanadyl sulafate or V(III)-cysteine complex. CONCLUSIONS: Vanadyl sulfate, cysteine and V(III)-cysteine complex exerted antitumor effects in tumor bearing rats. The V(III)-cysteine complex, however, exerts much more potent effects, as evident from the results of the present study. These beneficial effects of the above complex, in combination with its low toxicity provide evidence suggest its possible application in the treatment of human malignant diseases. PMID- 9858948 TI - Somatostatin analogue octreotide and melatonin inhibit bromodeoxyuridine incorporation into cell nuclei and enhance apoptosis in the transplantable murine colon 38 cancer. AB - There is much evidence of the antiproliferative activity of somatostatin (SS) and melatonin (Mel) upon the normal and neoplastic tissues. It has also been found, that both substances are able to alter, under certain conditions, apoptotic processes. Recently, it has been postulated that apoptosis plays a pivotal role in the control of tumour growth. So far, there is no data about the effect of SS analogue--octreotide (Sandostatin, SMS) and Mel on the apoptosis of colon cancer cells. The aim of this study is to examine the effects of SMS and Mel administered separately or together on apoptosis, bromodeoxyuridine incorporation and weight of tumours in the murine transplantable Colon 38 cancer. The male mice were implanted subcutaneously (s.c.) with a suspension of Colon 38 cells. After 6 days, the animals were subcutaneously injected with SMS, Mel, SMS and Mel together (once daily at 6-8 p.m., for 6 days). The incorporation of bromodeoxyuridine (BrDU) into cell nuclei was used as an index of cell proliferation (labelling index-LI). The in situ labelling of nuclear DNA fragmentation according to TUNEL method was considered as an apoptotic index (AI). Given separately, both SMS and Mel significantly decreased the LI and increased the AI. However, we have not observed any additive effect of SMS and Mel on either BrDU incorporation or apoptosis. The mean AI in the group treated jointly with SMS and Mel was significantly lower than in groups treated separately with SMS or Mel. It was also found, that the proliferation/apoptosis ratio were significantly lower in the group treated with SMS or MEL, which means that the imbalance between these two processes changed in favour of cell death. Possibly, the observed antitumour effects of these two substances could be due to this alteration. PMID- 9858949 TI - Over-expression of endoglin (CD105): a marker of breast carcinoma-induced neo vascularization. AB - The commencement of the complex process of carcinogenesis, and subsequent, rapid tumor growth and progression of mammalian neoplasms, including breast carcinomas (BCs), depends upon the continuous de novo formation of capillaries [i.e. neovascularization (NV)/neoplasm-related angiogenesis (NRA)]. The generation of a malignant, invasive cellular immunophenotype (CIP) and distant metastases, as aspects of tumor progression, are also NRA-dependent processes. Endothelial cells undergo rapid proliferation during mammary carcinoma-related angiogenesis. Human endoglin (CD105/EDG), is a homodimeric cell surface component of the transforming growth factor-beta (TGF-beta) type I receptor complex and is also a proliferation associated antigen (PM) expressed at high density on endothelial cells. Formalin fixed, paraffin-wax embedded, tissue sections (3-5 microns thick) of 15 BCs were employed for the assessment of EDG expression. An indirect, four-step, alkaline phosphatase (AP) (or diamino-benzidine [DAB]) conjugated, biotin-streptavidin based, antigen detection technique, employing the SN6h anti-EDG monoclonal antibody was conducted. Zymed's Histogold System was also utilized for immunocytological antigen detection. Strong expression (A; ++ + to ++ ++) of EDG on endothelial cells was demonstrated in all 15 BC cases. The most striking feature of the newly formed neoplasm-related capillaries was the presence of an enlarged perivascular space. Blood vessels in several normal human tissues (cortex, cerebellum, thymus, tonsil, spleen, lymph node, skin) used as control tissues contained significantly lower levels of EDG (B and mostly C; +/- to +), in accordance with the extremely slow turnover rate of normal endothelial cells. Furthermore, a close apposition between the capillaries and the adjacent parenchyma was observed in these normal controls. BCs, as most mammalian neoplasms, are characterized by extensive neovascularization and thus are candidates for anti-angiogenic therapy. Further studies should substantiate the importance of EDG expression in the earliest possible detection, diagnosis and NRA inhibition-based treatment of solid tumors, including BCs. PMID- 9858950 TI - [Value of orally administered retard morphine for therapy of severe pulmonary emphysema of the pink-puffer type. A pilot study]. AB - BACKGROUND AND OBJECTIVE: In severe "pink puffer" emphysema the patients' physical capacity is limited by dyspnoea despite maximal application of established treatment. This pilot study investigated the effect of retarded morphine, taken orally for 10 days, on ventilation, dyspnoea, walking capacity and wakefulness. PATIENTS AND METHODS: Twenty clinically stable patients (11 men, 9 women, mean age 68.5 [50-81] years) with "pink puffer" emphysema were studied over a period of 10 days in a prospective, non-controlled trial of cross-over design. Criteria for inclusion in the study were: 1-second forced expiratory volume (FEV1) < 1 I, vital capacity < 50% and normocapnia. In addition to their existing therapy patients received either no further therapy or retarded morphine. Morphine dosage was increased to maximally 3 x 30 mg daily, depending on effectiveness and side effects, dyspnoea at rest and immediately after a 6-min walk (assessed with Borg's visual analog scale), maximal walking capacity were determined, as well as blood gases, respiratory minute volume and the respiratory drive (airway occlusion pressure [P0.1]), responsiveness of the respiratory pathways to CO2 and wakefulness (concentration, fatigue, interest in surroundings). RESULTS: Twelve patients completed the study (group A). In the remaining patients (group B) the test had to be stopped prematurely because of undesirable side effects or an exacerbation of the underlying infection. In group A, morphine (mean dosage: 49.2 +/- 28.4 mg/d) caused a reduction of PaO2, dyspnoea on activity, the resting minute respiratory volume, respiratory drive and CO2 response, and an increase in PaCO2, HCO3- and the 6-min walking distance. Morphine did not produce a change in subjectively evaluated vigilance and the blood pH. CONCLUSION: After strict patient selection oral morphine produced a reduction of exercise dyspnoea and an increase in walking capacity in half of the patients with severe pulmonary emphysema. There also occurred a slight rise in PaCO2 without any relevant respiratory acidosis or significant decrease in wakefulness. PMID- 9858951 TI - [Endosonography controlled transgastric drainage of pancreatic pseudocysts]. AB - BACKGROUND AND OBJECTIVE: Endoscopic drainage of a pancreatic pseudocyst is an alternative to surgical intervention. But transmural drainage carries the risk of bleeding or perforation. Effectiveness and complication rate of endoscopic ultrasound-guided drainage, to avoid these risks, was investigated. PATIENTS AND METHODS: Eleven patients (eight men, three women; mean age 55 years) with a pancreatic pseudocyst (nine with alcoholic and two with biliary pancreatitis) were studied prospectively between 1996 and 1998. In all of them transpapillary drainage of the cyst had not been technically possible. After an endoscopic ultrasound (EUS) examination, the gastric wall was incised with a fistulotome under EUS guidance. A guide-wire was then advanced through the fistulotome into the pseudocyst. A double pigtail catheter was implanted for drainage. The size of the pseudocyst was monitored sonographically at two-week intervals. RESULTS: A cystogastrostomy was successfully established in ten of the twelve patients without serious complication. The pseudocyst was no longer demonstrated after a mean of 4.2 months (2 weeks to 6 months), while a small pseudocyst (1.6 cm [0.9 2.4 cm) remained in three patients. In two of the latter the size of the pseudocyst increased again after removal of the drainage catheter. Complete drainage by repeat cystogastrostomy succeeded in one of them, while a cystojejunostomy was established in the other. CONCLUSION: Endoscopic ultrasound guided transgastric drainage of a pancreatic pseudocyst is an effective treatment with few complications. PMID- 9858952 TI - [Syncope in 3rd degree atrioventricular block. Detection of virus genome in the myocardium]. AB - HISTORY AND CLINICAL FINDINGS: A 28-year-old woman was admitted after syncope which had been preceded by several flulike episodes. There was no history of any other serious disease. Physical examination was unremarkable. Heart sounds were regular and normal, there were no murmurs. INVESTIGATIONS: White cell count was 9400/microliter, with a normal differential count. Erythrocyte sedimentation rate and C-reactive protein were also normal. Virus serology revealed no abnormality. The electrocardiogram (ECG) showed complete (third degree) atrioventricular (AV) block with an idioventricular rhythm of 38 beats/min and right bundle branch block pattern. TREATMENT AND COURSE: A temporary transvenous pacemaker was inserted on the first hospital day. As myocarditis was suspected a right ventricular endomyocardial biopsy was obtained. Histological and immunohistological examinations demonstrated no unequivocal findings. But molecular-biological tests revealed. Coxsackie-B3 virus genome. The pacemaker was removed on the 6th day, when the ECG had shown intermittent second degree AV block. Regular sinus rhythm with a PR interval of 0.18 s was recorded on day 12, and 24-hour ECG monitoring for several days until her discharge on the 18th day confirmed this rhythm throughout. CONCLUSION: In aetiologically undetermined disease molecular-biological techniques can be indispensable for the exact diagnosis and may be decisive for administering specific treatment. PMID- 9858953 TI - [Therapy of acute migraine attacks and prevention of migraine]. PMID- 9858954 TI - [Tissue transglutaminase as autoantigen in endemic sprue. New aspects in diagnosis and etiopathogenesis]. PMID- 9858955 TI - [HIV infection as an occupational disease in a hospital physician. Decision of the Federal Social Court 18 November 1997]. PMID- 9858956 TI - [Interferon in hepatitis B]. PMID- 9858957 TI - [Cartilage regeneration therapy by autologous cartilage cell transplants]. PMID- 9858958 TI - Community-acquired pneumonia. AB - Community-acquired pneumonia (CAP) is a significant cause of morbidity and mortality in all age groups, especially the elderly, which is a patient population that continues to grow. Recently the spectrum and clinical picture of pneumonia has been changing as a reflection of this aging population; this requires a reassessment of and a new approach to the patient with pneumonia. Currently, pneumonia patients are classified as having either community-acquired or hospital-acquired infection rather than typical versus atypical. Patients who have CAP are categorized by age, presence of a coexisting medical illness, and the severity of the pneumonia. The rationale behind categorizing patients is to stratify them in terms of mortality risk to help determine the location of therapy (e.g., outpatient, inpatient, intensive care unit) and focus the choice of initial antimicrobial therapy. Once the decision to hospitalize a patient with pneumonia is made, the next step is to decide on an appropriate diagnostic evaluation and antibiotic therapy. Both decisions have evolved over the last several years since the publication of the American Thoracic Society's CAP guidelines. The current approach to the diagnostic work-up of pneumonia stresses a limited role of diagnostic tests and procedures. The antimicrobial regimen has now evolved into one that is empiric in nature and based on the age of the patient, the presence of coexisting medical disease, and the overall severity of the pneumonia. This process is a dynamic once because bacterial resistance to commonly used antibiotics can further complicate the course of pneumonia therapy, but the impact of resistance on outcome is less clear. Resistance of Streptococcus pneumoniae to penicillin is a prime example of this growing problem, and adjustment to pneumonia therapy may be required. A difficult but not uncommon problem in pneumonia patients is slow recovery and delayed resolution of radiographic infiltrates. Factors that impact negatively on pneumonia resolution include advanced age and the presence of serious comorbid illnesses such as diabetes mellitus, renal disease, or chronic obstructive pulmonary disease. In addition, certain organism factors (e.g., intrinsic virulence) may interact with host factors and advanced age to delay pneumonia resolution. For example, 50% of patients with pneumococcal pneumonia have radiographic clearing at 5 weeks, and the majority clear within 2 to 3 months. Recent data demonstrate that radiographic resolution of CAP is most influenced by the number of lobes involved and the age of the patient. Radiographic clearance of CAP decreases by 20% per decade after age 20, and patients with multilobar infiltrates take longer to clear than those with unilobar disease. In general, when approaching slowly resolving infiltrates after pneumonia, bronchoscopic evaluation and lung biopsy are more likely to yield a specific diagnosis if the patient is a nonsmoker younger than 55 years old with multilobar disease. If the patients has either no identifiable factors associated with prolonged pneumonia resolution or the repeat chest radiograph at 1 month shows no appreciable change, further diagnostic testing is indicated. The route and duration of antibiotic therapy, another detail of the management of CAP patients that has changed recently, is complicated by the fact that the majority of patients with CAP have no pathogen identified. Therefore, in most instances the physician initiates empiric antibiotics on the basis of epidemiologic data. If an etiologic pathogen is identified (either initially or at a later time), then the antibiotic spectrum can be narrowed. When no pathogen is discovered, broad-spectrum empiric antibiotics are continued. (ABSTRACT TRUNCATED) PMID- 9858959 TI - [Compliance with regulations for medical records in 15 Aquitaine public health facilities. Aquitaine DIM Physician's Group]. AB - OBJECTIVE: Record keeping contributes to the quality of health care in addition to fulfilling medicolegal and accreditation regulations. We conducted an audit of patient files in 15 public hospitals in Aquitaine to determine their compliance with national laws. METHODS: One hundred patient files were randomly selected in each hospital. Their compliance with 18 criteria established in 1992 by the ANDEM was studied. These 18 criteria were updated in accordance with the latest decrees from the health ministry and precisely defined in order to achieve good reproducibility of the measurements. RESULTS: Ten percent of the files fulfilled all 18 criteria. Each component of the record was correctly identified and the name of the general practitioner, admission documents, and delivery, anesthesia and transfusion records were almost always present. Surgery reports were found in 65 to 89% of the files. Low compliance with three criteria was observed: authorization to give health care to a minor (39 to 80% of the files), identification of the physician who prescribed the drugs during hospitalization (6 to 32%) and discharge prescription (42 to 95%). We also pinpointed problems concerning archiving and retrieval, especially concerning the results of complementary exams which were not necessarily stored with the patient files. CONCLUSION: This audit allowed us to define the first steps of a quality improvement project for patient records. We were able to deduct recommendations for analyzing compliance with legal requirements and suggest that the 1992 ANDEM criteria be updated to allow for valid comparisons between health care establishment. PMID- 9858960 TI - [Aneurysm of the iliac artery in an elderly subject. Role of treatment with covered endoprosthesis]. AB - BACKGROUND: Coated stents can be an alternative to open surgery for arterial aneurysms, particularly in patients with high operative risk. CASE REPORT: A 90 year-old man with advanced stage cardiopathy developed a 52 mm aneurysm of the iliac artery. Perioperative imaging (arteriography with marked catheter, CT-scan and endovascular ultrasonography) provided a precise description. The material used (PTFE coated thermal memory endoprosthesis, balloon in the hypogastric) provided effective exclusion. DISCUSSION: Endovascular treatment can be a useful alternative to open surgery for arterial aneurysm in the very elderly. PMID- 9858961 TI - [Association of an epsilon wave and syncope]. AB - BACKGROUND: Epsilon waves, rarely observed in clinical practice, result from late potentials favoring the development of ventricular rhythm disorders by reentry. CASE REPORT: A 53-year-old man with sequellar myocardial infarction experienced a syncope. The surface ECG recorded an epsilon wave. Programmed ventricular stimulation before and after anti-arrhythmia drugs triggered ventricular tachycardia which was hemodynamically poorly tolerated. A defibrillator was implanted and confirmed retrospectively the rhythmic origin of the syncope. DISCUSSION: The association of an epsilon wave and syncope should guide the etiology search towards severe ventricular rhythm disorders such as ventricular tachycardia. An electophysiologic study is required in order to determine the appropriate therapy and thus help avoid possibly fatal recurrence. PMID- 9858962 TI - [Detection of fecal occult blood: comparison of immunologic with guaiac tests]. PMID- 9858963 TI - [Primary biliary cirrhosis revealed by autoimmune thrombopenic purpura]. PMID- 9858964 TI - [Plasmodium falciparum malaria. 4 years after returning from an endemic zone]. PMID- 9858965 TI - [Chronic pain and psychological distress: a universal causal relationship?]. PMID- 9858966 TI - ["Evidence-based medicine". Standardized medicine or promotion of critical spirit?]. PMID- 9858967 TI - [Implantable cardiac defibrillators]. PMID- 9858968 TI - [Interview with S. Levy. Apropos of recommendations on defibrillators. Interview by M. Herida]. PMID- 9858969 TI - [Mucocutaneous manifestations of dengue]. AB - CHANGING EPIDEMIOLOGY: During the last half of the 20th century, dengue has spread to most countries in the inter-tropical zone and today is the most common cause of human arbovirus infection. In tropical zones, dengue is a frequent cause of fever and can be observed in travellers returning from an endemic zone. SKIN AND MUCOSAL MANIFESTATIONS: Though variable, skin and mucosal manifestations are important to recognize. Exanthema occurs late after constitutional symptoms and is suggestive. Minimal hemorrhagic lesions involving the skin and mucosa are observed in common dengue. They usually do not signal progression to authentic hemorrhagic dengue or shock. DIAGNOSIS: Skin manifestations of dengue raise the more general problem of diagnosis in travellers returning from tropical zones who develop fever and rash. PMID- 9858970 TI - [Interaction between converting enzyme inhibitors and hypoglycemic sulfonamides or insulin]. AB - POSSIBLE INTERACTIONS: Converting enzyme inhibitors (CEI) can intensify the glucose lowering effect of insulin or sulfonureas. This interaction has been observed in a few clinical cases and studied systematically in regional pharmacovigilance centers. LOW RISK RATE: This interaction occurs in certain patients with well-controlled diabetes who are given a CEI or when CEI dosage is increased. The risk of hypoglycemia in diabetics treated with CEI is very low and can be considered as negligible compared with the benefit of this class of well tolerated renal-protective antihypertension agents. DRUG DEPENDENT: A larger number of cases of hypoglycemia have been reported with captopril and enalapril, probably because these two CEI have been marketed for a longer period. Captopril may have a stronger hypoglycemia effect than enalapril. It has been demonstrated with hyperinsulin euglycemic clamp that captopril improves sensitivity to insulin in healthy volunteers with normal blood pressure; with enalapril however, the increase in insulin sensitivity is less pronounced and non-significant. AN INDIRECT MECHANISM: Captopril, and to a lesser extent enalapril, indirectly increases insulin sensitivity by increasing circulating kinine which leads to vasodilatation in the muscles and increased glucose uptake in muscle tissue. PMID- 9858971 TI - [Purulent pleurisy and empyema. With the exception of pleural tuberculosis]. AB - CLINICAL SETTING: Purulent collections in the pleural cavity usually occur as complications of pneumonia in immunodeficient or socially underprivileged patients. PUNCTURE ASPIRATION: The key to diagnosis, pleural aspiration is indicated in case of sufficiently abundant collections, especially in patients with fever. Exploratory puncture is a therapeutic emergency, allowing optimal antibiotic therapy when a causal germ is isolated and drainage of the purulent collection. If there is the slightest doubt, imaging techniques should be used to guide the puncture. DRAINAGE: Drainage is essential and is indicated whenever the aspiration fluid is purulent, contains, germs or the chemistry suggests major bacterial colonisation (acid pH, low glucose, high lactic acid dehydrogenase). Local injections of fibrinolytic agents improve drainage. PROGNOSIS: Complete recovery without sequellae is usually achieved. Physical therapy, provided early and for a prolonged period, helps improve the prognosis. Early care reduces the risk of recurrence of this potentially severe condition. PMID- 9858972 TI - [Thrombosis of the abdominal aorta]. PMID- 9858973 TI - Effect of baffles and a louvered bypass on the airflow and the convective patterns of contaminant inside a fume hood. AB - Various numerical predictions are given for the containment testing of fume hoods with internal baffles of different size and at different locations and with a louvered bypass. The numerical predictions were compared with experimental data and a fairly good agreement for the air velocity was found, although there was only qualitative agreement for the concentration of the contaminant. It was found that the level of contaminant leakage at the working aperture depended not only on the size of the recirculating airflow behind the fume hood sash but also on the blockage caused by the baffles inside the fume hood near the exhaust duct. Finally, some new suggestions for the design of the fume hood have been proposed. However, before implementing these suggestions it is essential that either experimental testing be performed, or the cost-effective alternative of using numerical predictions for containment testing, to determine accurately the level of contaminant leakage from the working aperture. PMID- 9858974 TI - Determinants of wheat antigen and fungal alpha-amylase exposure in bakeries. AB - The study's objectives were to measure flour antigen exposure in bakeries and define the determinants of exposure. Ninety-six bakery workers, employed in seven different bakeries, participated in the study. Two side-by-side full-shift inhalable dust samples were obtained from each study participant on a single occasion. The flour antigen exposure was measured as wheat antigen and fungal alpha-amylase content of the water-soluble fraction of inhalable dust, assayed via enzyme-linked immunosorbent assays. During the entire sampling period bakers were observed and information on 14 different tasks was recorded at 15-minute intervals. Other production characteristics were also recorded for each sampling day and used in statistical modeling to identify significant predictors of exposure. The mean alpha-amylase antigen exposure was 22.0 ng/m3 (ranging from below the limit of detection of 0.1 ng/m3 to 307.1 ng/m3) and the mean wheat antigen exposure was 109 micrograms/m3 (ranging from below the limit of detection of 1 microgram/m3 to 1018 micrograms/m3). Regression models that explained 74% of variability in wheat antigen and alpha-amylase antigen exposures were constructed. The models indicated that tasks such as weighing, pouring, and operating dough-brakers increased flour antigen exposure, while packing and decorating resulted in lower exposures. Croissant, puff-pastry, and bread/bun production lines were associated with increased exposure, while cake production and substitution of dusting with the use of divider oil were associated with decreased exposure. Exposure levels can be reduced by the automation of forming tasks, alteration of tasks requiring pouring of flour, and changes to the types of products manufactured. PMID- 9858975 TI - Application of a tracer gas challenge with a human subject to investigate factors affecting the performance of laboratory fume hoods. AB - The results of a "user" tracer gas test were applied to investigate the effects of various parameters on hood containment ability and to evaluate accepted methods to classify hood performance. This user tracer gas test was performed with a human subject standing in front of the hood. Based on the data collected, face velocity, its variability, and cross drafts are important in determining hood leakage. Results indicate that the temporal variability of face velocity may deserve as much consideration as its spatial variability, a parameter more traditionally recognized as being important. The data collected indicate that hoods with horizontally sliding sash doors perform better with the doors positioned to provide a center opening rather than when all of the doors are pushed to one side. The observed smoke patterns suggest that this trend is caused by the location and instability of vortices formed along the perimeter edge when all doors are pushed to one side. The results of manikin tracer gas tests and the user tracer gas test are inconsistent, suggesting that more research is needed to determine how best to evaluate whether a hood protects its users. PMID- 9858976 TI - Airborne contaminant exposure control in a partitioned work environment by exhaust ventilation systems. AB - The evaluation of contaminant removal effectiveness can play a key role in a working environment quality investigation and in remediation efforts. In this study a computer simulation technique was developed for assessing the efficiency of contaminant removal within a partitioned working environment. To determine the effect of the physical parameters of each partitioned space on contaminant removal effectiveness, a relative contaminant concentration unit was used to show the simulation results. In addition, tracer gas techniques were adopted to validate the accuracy of the prediction model. A comparison of measured CO2 concentrations in a controlled environment chamber with the results of a simulation model is presented. This study investigated the partition configuration of work environment and environment parameters including openings operation, constant concentration, and exhaust air volume mode. Results indicated that variations of the above three parameters can produce great differences in overall ventilation performance. Through this type of study, the interaction of airflow and contaminant concentration between partitioned spaces can be understood in advance and adequate knowledge can be provided to maintain a high quality and healthy environment for workers. PMID- 9858977 TI - Field validation of passive monitors for the determination of employee exposures to methylene chloride in pharmaceutical production facilities. AB - A series of field evaluations was performed to estimate the overall uncertainty of three manufacturers' passive monitors (Assay Technology Model 541 and 546, 3M Model 3520, and SKC Model 575-001) to determine methylene chloride (MeCl2) concentrations. Area samples were exposed in a pharmaceutical production facility at five MeCl2 air concentrations for both permissible exposure limit (PEL) and short-term exposure limit (STEL) periods. A specially designed evaluation chamber was used to concurrently expose six of each type of passive monitor while concurrently collecting six active samples from locations surrounding the dosimeter array. The active samples were used to estimate the actual concentration during the evaluation period. The precision, bias, and overall uncertainty were estimated for each monitor type at concentrations bracketing the Occupational Safety and Health Administration proposed exposure limits. The actual MeCl2 concentrations for the PEL sampling periods ranged from 0.9 to 63 ppm. The pooled overall uncertainty results for all the passive monitors evaluated under PEL sampling conditions met the NIOSH accuracy recommendations. Pooled overall uncertainty for PEL evaluations for the monitors were: Assay Technology Model 546 +/- 17%; 3M +/- 13%; and SKC +/- 17%. Actual MeCl2 concentrations for the STEL sampling periods ranged from 14 to 357 ppm. Pooled overall uncertainty results for Assay Technology and 3M monitors evaluated under STEL sampling conditions met the NIOSH accuracy recommendations; however, the SKC passive monitor was slightly greater than the NIOSH recommendation. Pooled overall uncertainty for STEL evaluations for the monitors were: Assay Technology Model 541 +/- 18%; 3M +/- 16%; and SKC +/- 27%. PMID- 9858978 TI - [Effect of loading suit "Penguin" on human metabolism during movements]. AB - Additional energy expenses due to stretching of the elastic elements of anti loading suit (ALS) "Penguin" as a whole (shoulders-feet) or only its lower part (waist-feet) in the course of cyclic leg movements were measured in five female and five male volunteers. ALS design enabled tensometric monitoring of efforts applied to specific elastic elements, and total efforts applied to the shoulder or pelvic girdles separately. Energy spend were determined with the indirect calorimetric techniques from the data of the expired air analysis. Registered were electromyograms of m. longus spinae, femoral extensor (m. biceps femoris) and femoral flexor (m. rectus femoris), and m. gastrocnemius. On the first stage, bicycle ergometer was pedaled w/o loading with a frequency of 60 cycles/min. The next stage included testing by incremental loading in which pedaling ceased at the pulse rate of 150/min. Results of the experiments that did not require stretching elastic parts of the suit and in which the total strain effort made up 20 to 25 kg and 15 to 16 kg by males and females, respectively, were compared. It was ascertained that ALS enhanced metabolism during motion by 20 to 30%; however, there was no significant difference in energy expenses when loaded by the whole suit or only its lower part. PMID- 9858979 TI - [Methodologic aspects of body water kinetic dynamic studies]. AB - In studying the level of hydration and liquid phases (LPs) in a space mission there use the stable and/or radioactive isotopes. The investigations are unique, the methods are adequate but not adapted in full measure to the challenges of the problem under study. The methodical approaches to the study of the dynamics of water metabolism are not available. Repeated introduction of the markers for these purposes is not acceptable. Another problem associates with taking the markers orally. In this case, a concentration of the markers will depend on the absorption and excretion processes. Prior to, during and in the readaptation period these functions will be different, making a correlation of the accumulated data difficult. There advances a possible version of solving these problems, namely, to use for the dynamic studies the residual contents of the markers in the LPs after single injection. However, this approach calls for investigating the kinetics of markers in weightlessness or during its ground-based simulation. The kinetics of tritium water has been studied in 6 volunteers under conditions of the 5-day bedrest and in 9 healthy men during free motor activity. There determined the characteristics of marker kinetics in a healthy man during his routine living activities. Under bedrest conditions there have been noted slowing down of the rate and a decrease in the degree of marker accumulation in the body after its single injection, the shift of a period of relative stabilization of marker content in LP to the more late dates of experiment, slowing-down of the marker excretion rate from the body. PMID- 9858980 TI - [Problem of orthostatic intolerance in astronauts and perspectives for its pharmacological prevention]. AB - Double-blind placebo-controlled method was applied to study venoconstrictor agent Dihydroergotamine effects on orthostatic tolerance in 8 healthy male-volunteers after 6-hour exposure to antiorthostatic stress (-6 degrees). The drug was able to induce blood centralization and diuresis additional to those caused by the simulated microgravity per se and failed to prevent the orthostatic intolerance. In 6 subjects the studies were repeated with the occlusive cuffs used in order to prevent venous blood return from the vascular bed of the lower limbs during 0 gravity simulation. The occlusive cuffs were shown to eliminate the additional drug-induced blood centralization and diuresis. Orthostatic tolerance in this case occurred to be less affected than in the case of no countermeasure used or in the case when dihydroergotamine was used alone. Thus, it was shown that mechanical prevention of the drug-induced blood centralization from being realized in 0-gravity simulation must be considered as the requirement necessary to permit the drug to perform its beneficial effect on the orthostatic tolerance after the real or simulated weightlessness. PMID- 9858981 TI - [Effect of antiorthostatic pre-training on transcapillary turnover of fluids in extra- and intracranial vessels of rats in antiorthostatic hypokinesia]. AB - Dynamics of the transcapillary turnover of liquids in the brain and soft tissues of the head was studied in pre-trained small laboratory animals (rats) during antiorthostasis, and their controls. Training for antiorthostasis consisted of tail-suspension for 2 hours in the period of two weeks. The transcapillary turnover of liquids was determined based on the arteriovenous difference in blood density (AVBD). Blood density was measured with the equipment of Anton Paar K.G. (Austria). Rats in the horizontal position did not exhibit any apparent trend in the dynamics of blood transcapillary turnover. Blood drain from the interstitial space at the time of return of the antiorthostatic rats into the horizontal position was dependent on the length of antiorthostatis. Beginning from the fifth hour of tail suspension, changed AVBD sign was an indication of edema of the muzzle soft tissue. This phenomenon was not observed in the pre-trained rats. Similar results were obtained in the investigation of cerebral vessels AVBD. Hence, changes in the transcapillary turnover of liquids in cranium during antiorthostatic hypokinesia point to the dominance of liquid filtration into the extravascular space. Antiorthostatic pre-training precludes liquid deposition in the interstitial space of the brain and the cerebral soft tissue. PMID- 9858982 TI - [Carbohydrate adaptive substance for prophylaxis of immune disorders and correction of dysbacterioses]. AB - Effects of a new preparation, adaptokhit, on the intestinal microbiocenosis and some parameters of immune resistance were studied in Macaque rhesus during 13-day hypokinesia. Experimental animals were given adaptokhit as an alimentary supplement preventing dysbiotic consequences of the microecologic unbalance due to motor restraint. Results of the investigation showed certain distinctions in the intestinal microflora in the experimental and control animals as early as on day three of the experiment. Intestinal microflora of the experimental primates who daily consumed adaptokhit at 50 mg/kg of the body remained stable throughout the period of observation. In contrast, in their controls lactoflora was found to decrease in parallel to the growth of opportunistically pathogenic endobacteria including representatives of Proteus and Clostridia sp., and development of dysbacteriosis of categories II and III. Besides, in the experimental animals adaptokhit prevented decline in the proliferative activity of lymphocytes further into adaptation. Also, at the final stage of observation following canceling the preparation there were signs of normalization of the allergologic status of primates in the experimental group. Therefore, adaptokhit can be used as an alimentary supplement to strengthen the colony resistance of animals in extreme conditions. PMID- 9858983 TI - [Peculiarities of cosmonauts fauces flora]. AB - In the course of the work done the specific and quantitative composition of the streptococcal autoflora of the fauces of the cosmonauts and the members of backup drew was investigated. In populations of isolated microorganisms the non pathogenic streptococci have dominated among which S. salivarius prevailed. The same species has constantly been isolated in all the cosmonauts, pre- and postflight. Observation of the microflora state of the fauces at different stages of their professional activity made it possible to reveal the peculiarities of an individual dynamics in the number of nonpathogenic streptococci isolated from tampon depending on the participation of the test-subjects in the previous space missions. This is evidently a reflection of the effect of psychoemotional tension on the state of colonization resistance (CR) of the fauces mucosa and, as result of this, on its microflora. In turn, the occurrence of the individual species of conditionally-pathogenic streptococci after the mission points to a decrease in the CR under effect of unfavourable factors of space mission. PMID- 9858984 TI - [Immobilized catalase in water purification systems]. AB - The results of studies on producing the biocatalyst based on catalase immobilized in the fibers from triacetate are presented. The catalase producer is Penicillium fungus. Catalase was produced by precipitation with the use of ethyl alcohol from the cultural fluid with separate and unseparate mycelium. The highest activity of catalase in the cultural fluid is seen on the nutrient medium containing 4% of carbon source. For immobilization the water solution of enzyme was concentrated in the vacuum-rotor evaporator at temperature of 25 degrees C. The enzyme was included in the structure of fibers during the process of their formation. Of the fiber-producing polymers (cellulose triacetate, chlorine, polysulphone) the most enzymatic activity has the catalase-containing fibers derived from the cellulose triacetate, in this case, the fine fibers of biocatalyst have the higher specific activity. It is established that the fibers obtained by using catalase of microbiological origin possess high stability and their activity does not practically change in the aqueous environment. The unpurified catalase is one and a half higher than at purified catalase. Under laboratory conditions there turned out the experimental batches of fibers and there conducted their endurance tests. Catalase included in cellulase triacetate has effectively functioned during a period of 2 years purifying the distilled water containing 50 mg/l of hydrogen peroxide. PMID- 9858985 TI - [Survival of laboratory animals in argon-containing hypoxic gaseous environments]. AB - Present paper is aimed at assessing the effect of hypoxic gaseous mixtures intended for the use as the fire suppression agents within closed volumes on the basis of argon with addition of carbonic acid on the body of mammalia. Three experiments were performed on the male white laboratory Wistar rats. It is indicated that argon adding to the hypoxic mixtures containing 4-5% volume increases animal survivability as compared to analogous nitrogen-based mixtures. An addition of 4-8% volume of carbonic acid impairs the of animal condition and reduces the survival rate. There noted a pronounced negative effect of the increased temperature on the survival time in the hypoxic media. One can draw inference about the principal possibility of using argon for formation of the hypoxic gaseous mixtures possessing fire suppression properties. PMID- 9858986 TI - [Cytoembryologic studies of super dwarf wheat grown in "Svet" greenhouse in the ground-based experiments]. AB - The Project of scientific programs MIR/SHUTTLE and MIR/NASA was allowed for studying the productional, cytoembryological, morphological, biomechanical and other characteristics of superclub wheat on cultivation in the Svet greenhouse on board orbital complex. This work was aimed at studying the duration of the complete cycle of ontogenesis of wheat and its individual stages, the peculiarities of forming the reproductive organs, processes, fertilization and formation of the seed production while cultivating in the Svet greenhouse under terrestrial conditions. Superclub wheat has been the object of experimentation. On cultivation of superclub wheat in the Svet greenhouse at designated conditions it was found that the cycle duration "from seed to seed" was 90-97 days. The number of granules in the wheat-ears studied was quite low and ranged from 15 to 30%. Performed studies with applying the light microscopy have indicated that in superclub wheat the embryological processes occur in compliance with those regularities which are described for the other forms of soft wheat. PMID- 9858987 TI - [Method for determination of radiation effects in cosmonauts by board dosimeter readings made during orbital mission]. AB - A technique for calculating the SS MIR absorbed doses from the galactic cosmic rays, protons of the Earth's radiation belts and solar flares is described. Calculated daily doses for different compartments along the MIR main axis are compared with doses calculated for the location of board radiometer R-16 in various periods of solar activity. Maximal doses in MIR compartments (in the absence of phantom) were compared with the doses in referential depths of a spherical phantom representing the blood forming tissue, skin, enteric epithelium, and the absorbed dose in the dosimeter location. This comparison allows more precise estimation of radiation hazard to cosmonauts in space flight. PMID- 9858988 TI - [Development of the device for Japanese quail nestlings maintenance in microgravity]. AB - Postembryonic development of the nestlings in microgravity has revealed their capacity for orientation and stabilization of their position in space which called for developing the special devices. Under terrestrial laboratory conditions the facility for maintaining the nestlings aged up to 10 days has been developed and tested. The results of tests have indicated that despite the restricted volume of individual cages the nestlings can grow without significant variations from the norm as evidenced by the dynamics of nestlings body mass during 10 days and the amount of consumed food. PMID- 9858989 TI - [Features of disturbances of phospholipid metabolism in patients with mechanical jaundice]. PMID- 9858990 TI - [Optimal rigidity of the protein structure: three classes of rigidity in the eubacterial RecA protein family]. PMID- 9858991 TI - [Potentially active copy of a LINE-1 element in HeLa cells: identification of a new specific promoter]. PMID- 9858992 TI - [Features of development of male sex cells in mice subjected to the effect of chemical mutagens dipine and nitrosomethylurea (NMM) during the prenatal period]. PMID- 9858993 TI - [Change in calcium and potassium currents in nerve cells during separate and combined application of diltiazem and ethanol]. PMID- 9858994 TI - [Differential mechanisms of density regulation in multispecies communities of microorganisms]. PMID- 9858995 TI - [Molecular mechanisms of lipid metabolism disruptions in the white rat brain during acute edema and normalizing effects of low-energy infrared laser radiation]. PMID- 9858996 TI - [CD3+4-8--thymocytes produce a chemotactic factor for stem elements of the bone marrow]. PMID- 9858997 TI - [A lipopolysaccharide of photosynthesizing bacteria suppresses the inhibitory effect of endotoxin on the cytochrome P-450 system of C57BL/6 mice in vivo]. PMID- 9858998 TI - [Reaction of the leech neuron to synaptic activation of various frequencies upon exposure to ganglioside antiserum]. PMID- 9858999 TI - [Use of radioresistance as a method for evaluating species independence of closely-related forms using the example of hamsters of the genus Phodopus (Rodentia, Cricetinae)]. PMID- 9859000 TI - [Protection of biological membranes from oxidation: the redox-chain NAD(P)H cytochrome b5 and lipid-radical cycle]. PMID- 9859001 TI - [Molecular modeling of the transmembrane segment of Na+-K+-ATPase. A pentagonal model]. PMID- 9859002 TI - [Some regularities in the generation and development of the neoplastic process in the body]. PMID- 9859003 TI - [Antistressor effect of adaptation to a physical load: role of nitric oxide]. PMID- 9859004 TI - [Circadian changes in levels of free amino acids in rat retina, and regulation of them by melatonin]. PMID- 9859005 TI - [Verapamil suppresses the activity of potential-regulated Ca2+-channels and does not affect activity of Ca2+-channels, regulated by intracellular Ca2+-stores, in PC-12 cells]. PMID- 9859007 TI - [Disruption of heat-shock protein synthesis does not prevent normal development of stress reaction in Drosophila melanogaster]. PMID- 9859006 TI - [Evaluation of viability of human embryonal brain cells in homotransplantation]. PMID- 9859008 TI - [Effect of fractions of peptides (1-10 kDa) from tissues of hibernating squirrels and peptides kyotorphin and neokyotorphin on fish behavior]. PMID- 9859009 TI - [Transcraniovertebral microstimulation corrects disturbances in childhood cerebral palsy]. PMID- 9859010 TI - [Carbon dioxide--a natural regulator of free radical homeostasis at various steps of evolution]. PMID- 9859011 TI - [New cationic liposomes for transfecting eukaryotic cells]. PMID- 9859012 TI - [Activation of serotoninergic neurons by metabolic precursors of serotonin: increase in frequency and duration of action potentials]. PMID- 9859013 TI - [Level of exchanged chromosome aberrations from long-term low-dose irradiation]. PMID- 9859014 TI - [Bacteria of the Spiroplasma genus infect two-spot ladybugs (Adalia bipunctata L.) in Russia]. PMID- 9859015 TI - Understanding changes in sexual activity among young metropolitan men: 1979-1995. AB - CONTEXT: Changes in the sexual behavior of teenagers can have a significant impact on levels of adolescent pregnancy and transmission of sexually transmitted diseases (STDs). Understanding the role played by attitudes and educational efforts will provide critical prevention information. METHODS: Data on the sexual behavior, sexual attitudes, educational experiences and demographics of 2,087 never-married metropolitan males aged 17-19 from the 1979 National Survey of Young Men and the 1988 and 1995 waves of the National Survey of Adolescent Males were analyzed through multivariate methods to examine factors that predict sexual behavior as well as those that predict sexual attitudes. RESULTS: The percentage of males aged 17-19 who had ever had sex increased from 66% in 1979 to 76% in 1988 and then decreased to 68% in 1995. The frequency of sexual intercourse in the year prior to the survey increased significantly over time, although the lifetime number of sexual partners did not. Acceptance of premarital sex increased significantly from 1979 to 1988, then decreased significantly from 1988 to 1995. Over time, young men were increasingly likely to prefer having and supporting a baby to marriage, abortion or adoption as the resolution to a nonmarital pregnancy. Trends in attitudes were strongly associated with sexual behaviors, with more conservative attitudes predicting less sexual activity. AIDS education, which was nearly universal in 1995, was associated with decreased sexual activity, although not among black youths. CONCLUSIONS: More conservative sexual attitudes and increased exposure to AIDS education are key predictors of decreased sexual activity among adolescent males. However, broader societal factors, such as fear of AIDS and increased awareness of problems associated with teenage pregnancy and STDs, may underlie both attitudinal and behavioral changes. PMID- 9859016 TI - Abortion incidence and services in the United States, 1995-1996. AB - CONTEXT: In the 1980s, the number of abortion providers in the United States began to decline, and more recently, so has the number of abortions performed. Whether the decline in service providers, which was last documented in 1992, is continuing and whether this influences the availability and number of abortions is of public interest. METHODS: In 1997, the Alan Guttmacher Institute conducted its 12th survey of all known abortion providers in the United States. The number and location of abortion providers and abortions were tabulated for 1995 and 1996, and trends were calculated by comparing these data with those from earlier surveys. Limited data were also gathered on types of abortion procedures. RESULTS: Between 1992 and 1996, the number of abortions fell from 1,529,000 to 1,366,000, and the abortion rate decreased from 26 to 23 per 1,000 women aged 15 44. The number of providers fell 14%, to 2,042, with the greatest decline among hospitals and physicians' offices rather than clinics. Eighty-six percent of counties had no known abortion provider, and 32% of women aged 15-44 lived in these counties. Of the country's 320 metropolitan areas, 89 had no known abortion provider, and for an additional 12, fewer than 50 abortions each were reported. Seventy percent of abortions were performed in specialized clinics and only 7% in hospitals. In the first half of 1997, early medical abortions were being offered in about 160 facilities, virtually all of which were also providers of surgical abortions. CONCLUSIONS: While abortion services in some areas of the country have declined since 1992 and many women continue to have limited access to providers, other factors have probably had more influence on the level of abortions performed. Early medical abortion methods are too new to be a measurable factor in abortion access. PMID- 9859017 TI - Multiple sexual partners among U.S. adolescents and young adults. AB - CONTEXT: Because many teenagers and young adults fail to use condoms correctly and consistently, the number of sexual partners they have is an important risk factor for sexually transmitted diseases, including HIV. Identifying factors that are associated with having multiple partners can help in the design of disease interventions. METHODS: Data on 8,450 males and females aged 14-22 who participated in the 1992 Youth Risk Behavior Survey were used to examine the prevalence of and factors associated with young people's having multiple partners. RESULTS: In all, 63% of female respondents and 64% of males were sexually experienced. Among those who had had sex during the three months before the survey, 15% and 35%, respectively, had had two or more partners during that period. At each age, the majority of sexually experienced respondents had had more than one lifetime partner; between ages 14 and 21, the proportion who had had six or more rose from 8% to 31% among females and from 14% to 45% among males. In logistic regression analyses, alcohol use, illicit drug use and young age at first coitus were associated with increased odds that females had had two or more partners in the previous three months, and being married lowered the odds; black or Hispanic race or ethnicity, alcohol use and young age at first coitus increased the odds for males, and being married reduced the odds. As the number of reported alcohol-related behaviors increased, the adjusted proportion of respondents who had recently had multiple partners rose from 8% to 48% among females and from 23% to 61% among men. CONCLUSIONS: The strong association between alcohol use and having multiple sexual partners underscores the need to educate young people about the effects of alcohol on partner choice and the risk of infection with sexually transmitted diseases. PMID- 9859018 TI - Increased condom use among teenage males, 1988-1995: the role of attitudes. AB - CONTEXT: Understanding whether and to what degree changes in young men's attitudes explain increases in condom use over time can be useful in developing more effective disease prevention strategies. METHODS: Data from the 1988 and the 1995 National Survey of Adolescent Males are used to determine changes in attitudes toward condoms, pregnancy prevention and HIV and AIDS. Two-limit tobit models are employed to investigate the association between these attitudes and condom-use behavior and to examine how this relationship may have changed over time. RESULTS: Between 1988 and 1995, young men's attitudes toward partner appreciation of condom use, condom-use embarrassment and pleasure reduction from condom use all changed in a direction suggestive of more consistent condom use. However, attitudes related to pregnancy prevention and AIDS avoidance changed in a direction suggestive of less-consistent condom use. Changes over time in the strength of the relationship between three attitude measures (masculinity, pleasure reduction and partner appreciation) and condom use also were predictive of lower levels of condom-use consistency. Only the strength of the relationship between condom-use embarrassment and consistent condom use changed in a direction corresponding to observed increases in rates of condom use among young men. CONCLUSIONS: Many of the significant changes in young males' attitudes toward condoms do not explain the increase in consistent condom use among adolescent males that occurred between 1988 and 1995. However increasing male contraceptive responsibility and emphasizing the risks and consequences of contracting HIV appear to be viable routes for policymakers to explore. Efforts particularly need to be targeted toward Hispanics. PMID- 9859019 TI - State abortion policy, geographic access to abortion providers and changing family formation. AB - CONTEXT: One of the goals in cutting welfare payments and setting time limits on welfare receipt is the reduction of out-of-wedlock childbearing among poor women. Yet such changes may increase the demand for abortion at the same time that access to abortion has decreased, throwing into doubt the potential effect of these changes on the proportion of women who are heading families. METHODS: State and county fixed-effects models were used to estimate the effects of factors influencing abortion availability--geographic access, parental notification requirements and Medicaid funding restrictions--on the county-level proportion of women heading households. RESULTS: The decline in geographic access to abortion providers during the 1980s accounted for a small but significant portion of the rise in the percentage of women heading families (about 2%). Restrictions on Medicaid funding for abortion accounted for about half of the increase in female headship among blacks, while new state parental notification requirements contributed modestly to the rise in the proportion of white women heading single parent families. CONCLUSIONS: Welfare reform legislation and attempts to reduce the availability of abortion services in the United States appear to be working at cross-purposes. Cutbacks in access to abortion may have contributed modestly to the increase in the proportion of women heading households. PMID- 9859020 TI - Using pharmacies in Washington state to expand access to emergency contraception. PMID- 9859022 TI - Effect of growth hormone releasing hormone on luteinizing hormone stimulated progestin biosynthesis in cultured rat ovarian granulosa cells. AB - In the present study, we examined the effects of growth hormone releasing hormone (GHRH) on luteinizing hormone (LH)-stimulated progestin biosynthesis. Granulosa cells from pregnant mare serum gonadotropin (PMSG)-treated immature rats were cultured in vitro in the absence or presence of ovine. National Institute of Health LH (100 ng/ml) with various doses of GHRH (10(-9), 10(-8) and 10(-7) mol/l) for 24 h. At the end of the incubation period, the incubation media were collected and levels of progesterone, 20 alpha-hydroxypregn-4-en-3-one and cyclic adenosine 3',5'-monophosphate (cAMP) were measured. GHRH significantly stimulated progesterone production and cAMP accumulation in media in a dose-dependent manner in the basal state (p < 0.01 and p < 0.001, respectively). In the presence of LH, GHRH significantly inhibited LH-stimulated progesterone production in a dose dependent manner (p < 0.01), whereas increasing concentrations of GHRH produced progressive increases in cAMP accumulation (p < 0.05). Since increasing concentrations of GHRH produced progressive decreases in the ratio of progesterone to 20 alpha-hydroxypregn-4-en-3-one production in the LH-stimulated state (p < 0.01), GHRH may be involved in modulation of key steroidogenic steps concerned with progesterone degradation rather than formation in LH-stimulated rat granulosa cells. These results suggest that GHRH may regulate the effects of LH in granulosa cells and play an important role in the reproductive process. PMID- 9859021 TI - Serum insulin-like growth factor-I, insulin-like growth factor binding protein-3, sex steroids, osteocalcin and bone mineral density in male and female rats. AB - Although it has been reported that the rate of weight gain and linear growth increases markedly during puberty in rats, little is known about the relationship between endocrine changes and bone mineral density (BMD) changes upon sexual maturation in these animals. The aim of this study was to examine the levels of serum insulin-like growth factor-I (IGF-I), IGF binding protein (IGFBP)-3, sex steroids and osteocalcin, and the changes in BMD in normal aging male and female rats. Male rats exhibited increases in serum IGF-I and IGFBP-3 concentrations before increases in serum testosterone levels. IGF-I and testosterone peaked at 9 weeks of age, and thereafter remained in a steady state, whereas IGFBP-3 reached a peak at 7 weeks of age, and then gradually declined. A strong correlation between serum IGF-I and IGFBP-3 levels was found in subjects 3-9 weeks old. A highly significant correlation between serum IGF-I and testosterone levels was also found. In females, serum 17 beta-estradiol, IGF-I and IGFBP-3 levels increased gradually from 3 to 5 weeks old, peaked at 9 weeks, and then decreased slowly thereafter. The correlation coefficient between serum IGF-I and IGFBP-3 was highly significant. The correlation coefficient between serum IGF-I and 17 beta-estradiol levels was weak, although it was strongest when the subjects were 3-9 weeks old. Serum osteocalcin is a marker of bone formation; its level remained relatively high from 3 to 9 and from 3 to 7 weeks of age in males and females, respectively, although osteocalcin in both sexes declined gradually with age. As for bone mass, sharp increases in BMD in the tibia, femur and lumbar vertebrae appeared earlier in female than in male rats, and the BMD in females tended to be higher than in males between 5 and 9 weeks old. After 9 weeks of age, BMD in males was higher than that in females, as BMD in males continued to increase whereas females tended to remain in a steady state after this stage. The correlation coefficients between tibial BMD and serum IGF-I or IGFBP-3 levels were highly significant when the subjects were from 3 to 9 weeks old. Taken together, these results suggest that BMD development occurs earlier in female than in male rats. This sex-related difference in changes in the BMD pattern may result from the earlier onset of puberty in females, and from sex-specific differences in concentrations of IGF-I, IGFBP-3 and sex steroids during maturation. PMID- 9859023 TI - Enhancement of apoptotic susceptibility by interleukin-1 beta in human endometrial epithelial cells. AB - Interleukin-1 (IL-1) is considered to be an essential cytokine for embryonic implantation. Expression of the IL-1 receptor in endometrial luminal epithelium increases maximally in the peri-implantation period, and embryonic implantation in mice is blocked by the IL-1 receptor antagonist. However, the function of IL-1 alone in implantation is still unclear. It has been reported that endometrial epithelial cells undergo apoptosis at the implantation site. In this study we have investigated the regulatory function of IL-1 in endometrial epithelial apoptosis, using the human endometrial epithelial cell line HHUA, which is susceptible to Fas-mediated apoptosis. The enhancement of endometrial apoptosis by IL-1 beta was not accompanied by any increase in cell-surface expression of Fas, which suggested that IL-1 beta enhanced the postreceptor apoptotic signals in the activated cells. IL-1 may be a regulator of apoptotic susceptibility in endometrial epithelium in the peri-implantation period. PMID- 9859024 TI - Is there a role for leptin in human reproduction? AB - Leptin is a protein product from the obesity gene (ob gene). It has been shown that leptin significantly correlates with body mass index in humans. In contrast to the obesity of genetically obese (ob/ob) mice, human obesity is not generally caused by gene mutations. It is possible that human obesity results from central leptin resistance. Leptin can serve as a metabolic cue in the neuronal activation of gonadotropin releasing hormone (GnRH) at the end of the prepubertal period. The concentration of leptin is higher in pubertal girls than pubertal boys, and it is supposed that sexual dimorphism might be established in the prepubertal period or even in earlier developmental phases. This dimorphism could be explained by a suppressive action of androgens on leptin. Decreased leptin levels were found in undernourished women of reproductive age, mainly presenting with oligo- or amenorrhea. Leptin concentrations fluctuate according to the phase of the menstrual cycle. It is suggested that the complex relationship of leptin with other hormones, such as insulin, can have etiopathogenetic importance in some enigmatic reproductive disturbances such as the polycystic ovary syndrome. Recent findings of leptin in non-adipose tissue of the placenta could indicate its potential role in developmental physiology and human reproduction. PMID- 9859025 TI - Serum total renin levels after ovarian electrocautery in women with polycystic ovary syndrome. AB - Recent observations have suggested an enhanced activity of the ovarian renin angiotensin system in polycystic ovary syndrome (PCOS). Owing to technical restrictions, the direct measurement of ovarian renin-angiotensin activity is impossible. The measurement of total renin (active renin + prorenin) in serum is particularly valuable for analyzing the ovarian renin-angiotensin system, as 90% of circulating renin is in the form of prorenin and ovaries are the major extrarenal source of prorenin in females. Also, the renin synthesized by ovaries is in the form of prorenin. In the present study we hypothesized that ovarian trauma caused by electrocautery 'impairs' the activity of the ovarian renin angiotensin system, which in turn would interrupt the endocrine vicious cycle of PCOS, and restore normal ovarian function. To test this, we examined the effect of ovarian electrocautery on serum levels of total renin in 11 oligomenorrheic women, aged 25 to 36 years, with PCOS and anovulatory infertility. Against our basic hypothesis the serum total renin levels remained unaltered after ovarian electrocautery, while the serum levels of luteinizing hormone, testosterone and androstenedione declined. The mechanism that induces ovulation without altering total renin levels in serum remains to be resolved. PMID- 9859026 TI - Bone mineral density in premenopausal women receiving levothyroxine suppressive therapy. AB - Osteoporosis is a well-known complication of thyrotoxicosis. Prolonged subclinical hyperthyroidism due to L-thyroxine treatment has been associated with reduced bone mass and thus with the potential risk of premature development of osteoporosis. The aim of this study was to assess the effect of a chronic L thyroxine suppressive treatment on bone mineral density (BMD) in a group of premenopausal women. Forty consecutive patients (mean age +/- SE = 40.95 +/- 1.56 years) affected by non-toxic goiter underwent bone mineral densitometry (dual energy X-ray absorptiometry; DEXA) of the lumbar spine (L1-L4) and right femoral neck. At the time of the study the patients had been under thyroid stimulating hormone (TSH) suppressive therapy for 74.95 +/- 10.34 months (range 17-168 months). Baseline levels of free thyroxine (fT4), free triiodothyronine (fT3), TSH, calcium and phosphorus were measured and correlated with BMD. The age of starting, duration of treatment, main daily dose, cumulative dose of treatment and body mass index (BMI) were also correlated with BMD. Statistical analysis was performed by multiple linear regression. BMD among female patients was not significantly different from that of the general population matched for age and sex. With the use of the regression model, no significant correlation was found between BMD and the variables considered. In conclusion, our data suggest that L thyroxine suppressive therapy, if carefully carried out and monitored, has no significant effect on bone mass. PMID- 9859027 TI - High levels of serum allopregnanolone in women with premature ovarian failure. AB - The endocrine characteristics of patients with premature ovarian failure (POF) have not been fully elucidated. The aim of the present study was to evaluate whether steroidogenic activity in women with POF is different with respect to fertile and postmenopausal subjects. In particular, circulating levels of allopregnanolone, a neuroactive steroid involved in modulation of reproductive function in rats, have been evaluated and correlated with serum levels of delta 4 precursor (dehydroepiandrosterone sulfate (DHEAS)), delta 5 intermediates (androstenedione, 17-hydroxyprogesterone (17-OHP), progesterone) and final products (estradiol and testosterone) of androgens. Levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) and sex hormone binding globulin (SHBG) were also determined. In all cases specific radioimmunological assays were used. Women with POF showed statistically significantly lower concentrations of 17-OHP, androstenedione and testosterone when compared to fertile controls, while no differences were found between women with POF and postmenopausal women. Serum DHEAS levels were similar in POF patients and in fertile controls and higher with respect to postmenopausal women. Serum allopregnanolone levels were significantly higher in women with POF than in postmenopausal and in fertile women. A significant inverse correlation between allopregnanolone levels and menopausal age in patients with POF was observed while no significant correlation was found between allopregnanolone and progesterone, androstenedione, 17-OHP and testosterone levels. In conclusion, allopregnanolone is scarcely influenced by the reduction of ovarian and adrenal activity observed in POF. PMID- 9859028 TI - Virilizing ovarian tumor of cell tumor type not otherwise specified: a case report. AB - Whereas ovarian tumors with overt endocrine manifestations account for less than 5% of all ovarian neoplasms, the incidence of virilizing type tumors in postmenopausal women is even lower since the average age of occurrence is 43 years. Steroid cell tumors not otherwise specified (NOS) are even more rare. We report the case of a 56-year-old woman (age of onset of menopause 43 years) who consulted our service due to a hyperandrogenic syndrome: deepening of the voice, temporal balding, hirsutism and cliteromegaly. Laboratory findings indicated hyperandrogenism in male range. The dexamethasone suppression test did not modify basal values, indicating that adrenal origin was unlikely. Transvaginal ultrasound disclosed multiple microcysts in the left ovary. Abdominal tomography was normal. Suspecting an ovarian tumor, bilateral oophorectomy was performed and a pediculate, 3 cm in diameter, was encountered in the left ovary. Histopathological studies determined it to be a virilizing ovarian tumor NOS. Postoperative recovery was fast; normal hormonal values were reached together with visible clinical improvement. This case is reported because this type of tumor is very infrequent in postmenopausal women, and because in this case it was the functional hormonal test that allowed tumor localization. PMID- 9859030 TI - Trocars: safety and selection. Emergency Care Research Institute. AB - Trocars are used during laparoscopic procedures and other minimally invasive surgery (MIS) to make small, puncturelike incisions in outer tissue layers. These incisions allow surgeons to insert cannulas through which surgical instruments can be introduced. MIS procedures allow faster recovery times than traditional open procedures as well as minimal patient discomfort--but at the same time, they entail a certain amount of risk. Trocar-related injuries and complications range from minor bleeding to, on rare occasions, death. In this Guidance Article, we describe what trocars are, how they are used, and how these complications can be avoided. Our discussion includes the use of trocar tip shields, which are designed to reduce th risks of trocar use--but whose value is the subject of considerable debate among surgeons. It also covers the relative merits of reusable and disposable models. In addition, the article includes an overview of the trocar-cannula market, listing current suppliers and the types of trocars they offer. PMID- 9859029 TI - The psychotherapeutic effects of estrogens. AB - The effect of estrogens on the central nervous system, particularly mood and behavior, remains a controversial area which needs clarification, not just for understanding of depression in women but to ensure that such commonplace problems in women have efficient and appropriate therapy. There is now good evidence that estrogens are rapidly effective in the treatment of depression in many women but this information has not found its way through to those health care personnel, psychiatrists and psychologists who are principally involved in the treatment of depression. There is also strong evidence for the benefits of estrogens on cognitive functioning, not only in preventing the onset of dementia but also in improving the symptoms in the established condition. Recent work has also suggested a benefit for estrogens on mood in women diagnosed as suffering from chronic fatigue syndrome. This article reviews the effect of endogenous estrogen on the female central nervous system and the ever increasing evidence for the diverse psychotherapeutic effects of exogenous estrogens. PMID- 9859031 TI - Fire hazard created by the misuse of DuraPrep solution. PMID- 9859032 TI - Carbon monoxide exposures during inhalation anesthesia: the interaction between halogenated anesthetic agents and carbon dioxide absorbents. PMID- 9859033 TI - Undetected venous line needle dislodgment during hemodialysis. PMID- 9859034 TI - [Modern aspects in the management of acquired heart valve lesions]. PMID- 9859035 TI - Acquired heart valve pathology. An update for the millennium. AB - A survey of 200 surgically excised valves in 1996, from a single academic center, revealed that degenerative disease is by far the most common type of underlying pathology, both for the mitral valve and the aortic valve. This observation relates also to the fact that the vast majority of patients was 60 years or older. Post-inflammatory (rheumatic) disease was relatively rare; younger individuals with post-inflammatory valve pathology usually originated from parts in the world known to be endemic for rheumatic fever. The results further emphasize the shift in health costs related to an aging population. PMID- 9859036 TI - [Diagnosis and differential therapy of mitral stenosis]. AB - Clinical symptoms and diagnostic findings in patients with mitral stenosis are usually determined by the extent of the stenosis. Compared to a normal mitral valve area (MVA) of > 4 cm2, MVA in patients with severe mitral stenosis is usually reduced to < 1.5 cm2. In older patients symptoms are frequently influenced by concomitant diseases (e.g. atrial fibrillation, arterial hypertension or lung disease). An important diagnostic element besides anamnesis, auscultation, ECG and chest X-ray is echocardiography, which is required in order to measure non-invasively and reliably the mitral valve gradient (MVG), the MVA and morphologic changes to the valves, as well as concomitant valvular disease, ventricular functions and, where appropriate, left-atrial thrombi. In addition to the surgical treatment of patients with severe mitral stenosis, which has been an established procedure for 50 years, percutaneous balloon mitral valvuloplasty (MVP) has recently established itself as an alternative option. At the current time, the Inoue technique seems to display the most advantages. Following transseptal puncture, the Inoue balloon is guided transvenously into the left atrium and then into the left ventricle using a special support wire. The balloon is short and soft. Its special unfolding character enables it to be placed securely in the mitral valve without any risk of ventricular perforation (Figure 1). As with surgical commissurotomy, balloon valvuloplasty leads to a separation of fused commissures. This results in a significant reduction of MVG, accompanied by an increase in the MVA (Figure 2). The results and success of MVP are influenced by the morphology of the valves and the changes to the subvalvular apparatus. In randomized studies, the results of surgical commissurotomy were comparable with those of balloon mitral valvulotomy. In our hospital, an increase in MVA from 1.0 to 1.8 cm2 could be achieved in 899 patients (mean age 56 +/- 3 years). In younger patients with less significantly changed valves, the results were correspondingly more favorable than in older patients (Figure 3). Provided valve morphology is suitable, a relapse following previous surgical commissurotomy is not a contraindication for MVP. The MVP complication rate is very low in skilled hands: mortality is below 1%; mitral insufficiency occurs in 3 to 10% of interventions; we observed a severe mitral insufficiency in 5% of our patient group. Thromboembolic complications may be prevented after exclusion of atrial thrombi by transesophageal echocardiography. The occurrence of a hemodynamically significant atrial septum defect is a very rare event. The mid term results (5 to 10 years) and the low restenosis rate following MVP in patients with suitable valves are comparable with those of surgical commissurotomy. In older patients with considerably changed, calcified and fibrotic valves, restenosis may be expected within 1 to 5 years. In these patients MVP represents no more than a palliative intervention in order to prolong the point of surgery, for example in patients where a concomitant aortic valve disease in itself is not yet an indication for surgery. Special indications are to be found in young patients with severe mitral stenosis yet few symptoms, in pregnant females and in emergency situations, as well as in patients with Grade II mitral stenosis with intermittent atrial fibrillation. Catheter therapy is much less invasive than surgery. In case of failure the patient still has the option of surgical therapy. Patients with morphologically significantly altered valves usually receive a valve replacement since an unsuccessful reconstruction would lead to a second operation within a very short time interval. Contraindications for MVP are thrombi in the left atrium, a previously existing > Grade II mitral regurgitation and marked, degenerative destruction of the subvalvular apparatus or extensive calcification of the valves. MVP thus represents a significant addi PMID- 9859038 TI - [Diagnostic approach and optimal treatment of aortic valve stenosis]. AB - The slow progression of valvular aortic stenosis enables the left ventricular myocardium to adapt itself to the increasing afterload. When myocardial adaption is exhausted, surgical intervention is urgent, the prognosis, however, is already limited. To quantify the hemodynamic severity of aortic stenosis, transaortic pressure gradients (dp) measured by Doppler echocardiography or hemodynamically are inappropriate, because dp is significantly dependent on the transaortic flow volume. In severe aortic stenosis, despite constant narrowing of the aortic valve area, the reduced stroke volume results in decreasing transaortic pressure gradients. With aortic valve resistance or transaortic pressure loss (PL)--the quotient of pressure gradient and stroke volume--the hemodynamic severity of aortic stenosis can be described accurately. If PL is known, a decompensated aortic stenosis (PL > 1 mm Hg/ml) may be differentiated from myocardial failure of another etiology and a concomitant left ventricular outflow tract obstruction. With respect to medical therapy, the prevention of bacterial endocarditis and thromboembolic complications is important. Knowing the potential danger of syncopies and ventricular arrhythmias during exercise with increasing severity of aortic stenosis, patients have to be informed about their limited functional capacity. The occurrence of typical symptoms during the natural history of chronic aortic stenosis (e.g. dizziness, syncopes, angina pectoris, arrhythmias) manifestation of ST-T-alterations or silent myocardial ischemias and demonstration of an inadequate myocardial adaptation to the chronic pressure overload in asymptomatic patients are accepted indications for a surgical intervention. If the indication for surgery remains uncertain, stress tests (e.g. radionuclidventriculography) may be performed to demonstrate an exhausted myocardial adaptation. If the PL and the severity of aortic valve/anulus calcification is known, the progression of a chronic aortic stenosis can be estimated. This might be important, if a cardiosurgical intervention has to be performed for other indications and aortic stenosis is co-existent but does not require an intervention at that time. For prognostic reasons myocardial decompensation due to aortic stenosis is an indication for an urgent surgical intervention. Attempts for medical recompensation or bridging strategies (e.g. balloon valvotomy) worsens the prognosis significantly. PMID- 9859037 TI - [Optimal management of primary and secondary mitral regurgitation]. AB - While morphologic alteration of parts of the mitral valve apparatus (ventricular wall, papillary muscles, chordae tendineae, valve annulus and leaflets) may result in a loss of its functional integrity (primary mitral regurgitation, MR) mitral annulus dilatation following left ventricular enlargement or change in chamber geometry and consecutive opening of the angle between papillary muscles and valve annulus cause secondary MR. Irrespective of these etiologies MR is chronically progressive and much more than the severity of MR the grade of myocardial adaptation to the chronic volume overload is of prognostic significance. Inadequate myocardial adaptation is demonstrated by an increase of the echocardiographically determined radius (r) to wall thickness (Th) ratio (r/Th > 3.0), indicating increasing left ventricular wall stress or by an insufficient increase of the left ventricular ejection fraction (< or = 5% of resting values) under exercise conditions, e.g. with radionuclide angiocardiography (RNV). Stressecho may replace RNV in the future for this indication. Actually, stress echo is not reliable to determine changes in left ventricular ejection fraction at rest versus exercise because of systematic errors and error reproduction. There are preliminary reports on biochemical markers like noradrenaline or tumor necrosis factor alpha being helpful to determine the breakdown of myocardial adaptation mechanisms. Surgical intervention is indicated in chronic MR irrespective of the hemodynamic severity, if myocardial adaptation is inadequate. If mitral reconstruction, the surgical technique of choice, remains insufficient to restore normal valve function, mitral valve replacement with preservation of the subvalvular apparatus is unavoidable. For a deceleration of the progressive volume overload in chronic MR for which a surgical intervention is not yet indicated, a long-term afterload reducing medical therapy preferably with long acting ACE-inhibitors seem to be prognostically favorable. PMID- 9859039 TI - [Diagnosis and indication for aortic valve replacement in asymptomatic and symptomatic patients with aortic regurgitation]. AB - Chronic volume overload is associated with dilatation and eccentric hypertrophy of the left ventricle (= ventricular remodeling). With the dilatation of the left ventricle and the shift of the pressure-volume-relationship to the right, the filling pressures can be kept normal despite severe regurgitation. Therefore, the patient with aortic regurgitation can remain asymptomatic over many years. Thus, the indication for aortic valve replacement in patients with severe aortic regurgitation is sometimes difficult and may lead to problems to choose the optimal time point for operation. As a general rule, symptomatic patients with severe aortic regurgitation should be operated as soon as possible. In asymptomatic patients with significant dilatation of the left ventricle and reduction of systolic pump function the therapy of choice is aortic valve replacement. Asymptomatic patients with normal left ventricular function have usually a good prognosis with a yearly mortality rate of approximately 0.04%. However, in the presence of significant dilatation of the left ventricle, i.e. enddiastolic chamber diameter more than 70 mm respectively endsystolic diameter more than 50 mm, patients have to be checked on a regular basis, i.e. in yearly intervals to detect left ventricular dysfunction in due time. According to the literature, asymptomatic patients with severe aortic regurgitation develop left ventricular dysfunction in a yearly rate of 4%. However, approximately 50% of all patients are even after 10 years asymptomatic. The indication for aortic valve replacement is given when the patient shows a deterioration of left ventricular function or becomes symptomatic. Valve replacement is also indicated in patients with an ejection fraction below 50% and/or endsytolic chamber diameter of more than 55 mm. Therapy of choice in symptomatic patients with severe aortic regurgitation is aortic valve replacement. In asymptomatic patients, operation depends on the degree of chamber dilatation respectively the severity of left ventricular dysfunction. In patients with severe aortic regurgitation but without clinical symptoms and moderate enlargement of the left ventricle regular check ups in yearly intervals are indicated. In the presence of severe left ventricular dilatation check-ups should be performed on a half-year basis to prevent irreversible damage to the heart muscle. PMID- 9859040 TI - [Prevention and management of dysfunction of prosthetic heart valves]. AB - The ideal heart valve prosthesis has not been developed. The today available mechanical and biological prostheses are far from perfect concerning thrombogenesis, mechanical durability and hemodynamic performance. Failure of an implanted valve is possible due to inadequate indication, inadequate surgical technique and inadequate medical follow-up. The patient with a heart valve prosthesis must be seen as a life-long challenge for the cooperation between cardiologists and cardiac surgeons. This article describes the main causes of prosthetic heart valve dysfunction and the surgical treatment, including structural dysfunction, thrombosis, paravalvular leakage, tissue ingrowth and prosthetic valve endocarditis. PMID- 9859041 TI - [Artificial bradycardias due to tape-running problems of old long-term ECG records]. AB - Artificial bradycardias due to tape-running alterations of old long-term ECG recorders were observed with increasing frequency in the last 2 years at our hospital. To document the extent of this problem, 115 consecutive 24-hour long term ECG recordings of 10 still used tape recorders with an age of 12 +/- 4 years were examined. Analysis of the tapes with a new analysis system revealed artificial bradycardias with rates of 10 to 40/min in 22 of 115 long-term ECG recordings (19%). These artificial bradycardias were observed in 4 out of 10 examined tape recorders (40%). Three of 4 concerned recorders were 14 years old at the time of examination, and one recorder was only 5 years old. Artificial bradycardias were caused by extensive alterations of tape-running speed in all cases. Diagnostic proof of artificial bradycardias due to tape running alterations on long-term ECG are simultaneous with prolonged RR-intervals occurring increases in all ECG times including P-width. PQ-time, QRS-width and QT duration with otherwise unchanged ECG morphology. To avoid prolonged hospital stays or therapeutic mistakes like unnecessary pacemaker implantation, especially older long-term ECG-tape recorders should be checked regularly and, if necessary, be replaced by new devices. PMID- 9859042 TI - [Therapy recommendations for adverse effects of Viagra and nitrates (NO-donors)]. PMID- 9859043 TI - [Safe and unsafe indications to the electrophysiological examination]. PMID- 9859044 TI - [Systemic cytostatic chemotherapy]. PMID- 9859045 TI - [High dose therapy with stem cell transplantation]. PMID- 9859046 TI - [Endocrine therapy of solid tumors]. PMID- 9859047 TI - [Immunotherapy: cytokines and tumor cell vaccines]. PMID- 9859048 TI - [Locoregional therapy methods]. PMID- 9859049 TI - [Endoscopic therapy of solid tumors of the gastrointestinal tract]. PMID- 9859050 TI - [Unconventional, alternative therapy methods in oncology]. PMID- 9859051 TI - [Myelodysplastic syndromes]. PMID- 9859053 TI - [Percutaneous endoscopic gastrostomy in ventriculoperitoneal shunt]. PMID- 9859052 TI - [Secondary hypersplenism syndrome. Etiology of thrombo- and leukocytopenia in sarcoidosis?]. PMID- 9859054 TI - [Acetylsalicylic acid in renal failure]. PMID- 9859055 TI - [Sling biopsy in gastroscopy]. PMID- 9859056 TI - [The 13C urea breath test]. PMID- 9859057 TI - ["Routine laboratory screen". Cost saving in the hospital laboratory]. PMID- 9859058 TI - [Determination of coronary flow reserve]. PMID- 9859059 TI - [Acute dyspnea]. PMID- 9859061 TI - [Legal protection from competition clause]. PMID- 9859060 TI - [Acute dyspnea]. PMID- 9859062 TI - [Vaccination after splenectomy and in asplenia]. PMID- 9859063 TI - [Incidence of de novo neoplasms after liver transplantation]. AB - BACKGROUND: De novo malignancy developing after transplantation constitutes a well-known complication or organ transplantation, mainly described among renal recipients. AIM: To determine the incidence of de novo internal malignancies (excluding therefore skin cancers and recurrent hepatocellular carcinoma) in a cohort of 183 patients undergoing liver transplantation (OLT) between 6/1/1991 and 12/1/1996 with a minimum follow-up of 12 months and under cyclosporine azathioprine-prednisone. PATIENTS AND METHODS: The study comprised 183 patients (mean age: 53 [8] years, with 70% males) whose charts were reviewed retrospectively. Rejection, steroids treatment, methyl-prednisolone bolus and OKT3 use were compared in the cases and in the matched control group. RESULTS: Seven malignancies were detected: 3 lung carcinomas, 1 larynx, 1 prostate, 1 colon and 1 ovarian. The mean age was 53 (8) years. The diagnosis of cancer was made at an average time of 24 (17) months (range, 10-54) post-OLT. Three patients died with a mean survival of 31 (16) months. OLT indication was mainly for viral liver disease (5/7). Although not statistically significant, immunosuppression data were higher among patients with cancer than in the matched group. Two additional patients developed post-transplantation lymphoproliferative diseases at 2 and 9 months, respectively. CONCLUSION: We observed a wide variety of malignancies after OLT, but no associated factor was found, even though there was a trend to higher doses of immunosuppression in patients with cancers. The institution of preventive measures and surveillance programs may allow for early institution of therapy, improving therefore the survival. PMID- 9859064 TI - [Bone mass in patients with cystic fibrosis of the pancreas. Relationship with anthropometric parameters and genotype]. AB - BACKGROUND: Cross-sectional study of bone mineral density (BMD) in children and adolescents with cystic fibrosis of the pancreas. The relationship of BMD values with nutritional status, respiratory function and the cystic transmembrane regulator genotype was also evaluated. PATIENTS AND METHODS: BMD expressed as grams of hydroxyapatite/cm2 was measured by dual-energy X-ray absorptiometry in the lumbar spine (L2-L4) in 41 patients (21 males and 20 women; age range: 4-21 years) with cystic fibrosis of the pancreas and compared with that of 471 normal controls (256 males and 215 women; age range: 1-20 years). Twenty patients were prepubertal, 9 pubertal and 12 young adults. RESULTS: Clinical repercussion of the disease evaluated by clinical and anthropometric data (weight, height and body mass index) and respiratory function was considered moderate. Height z score (mean [MSE]) was -0.53 (0.28), weight -0.81 (0.21) and body mass index -0.82 (0.12) BMD z score values (mean [MSE]) were -1.14 (0.17) and differed significantly (p < 0.001) from those of normal age- and sex-matched controls. No significant differences were observed between males and women or among prepubertal, pubertal and young adult patients. BMD z score values less than-1 z score were found in 53% and under -2 z score in 8%. Cystic transmembrane regulator genotype was studied in 36 patients (17 were F508/-, 10 F508/F508, 5 G542X/- and 4 diverse) and did not predict bone mineral status. A statistically significant correlation was found between BMD z score values and height z score, weight z score, body mass index z score and clinical assessment according to Shwachman criteria. A negative and statistically significant correlation was observed between BMD z score and functional score. CONCLUSIONS: The decrease in BMD values in CF patients begins early in life and appears to be related to the degree of clinical expression of the disease. PMID- 9859066 TI - [Continued training: a difficult dilemma]. PMID- 9859065 TI - [Scientific activity of the most productive Spanish research teams in pharmacology and pharmacy during the period 1986-1993 as covered by the Science Citation Index (SCI)]. AB - BACKGROUND: Bibliometric analyses at the research team level in the pharmacology and pharmacy (F&F) subfield allow in-depth analysis of the findings obtained at higher aggregation levels. MATERIAL AND METHODS: Publications of Spanish authors in pharmacological journals covered by the Science Citation Index (SCI) are studied and the most productive Spanish teams during 1986-1989 and 1990-1993 are identified through in-house programmes based on co-authorship analysis. Team composition is defined and team size, scientific output, productivity, expected impact factor and collaboration pattern of the most productive teams are analysed as well as its evolution over time. RESULTS: The observed annual increasing rate in the number of publications and authors in the field (11%) was similar to that of the number of researchers in the Spanish scientific system (8%). The number of highly productive teams doubled from the first to the second period (28 and 61 teams, respectively). The average team size was 8 researchers, showing an average scientific output of 3 documents/year and an average, productivity of 1.7 publications per author and team in each period. Over the years, an increase in the expected impact factor (EIF) of the publication journals and significant changes in the team composition were observed. A total of 13 stable teams were identified, with a greater size and output than the rest of the groups, and showing an upward trend in their expected impact factor and international collaboration rate. Stable teams did not show the negative correlation between team size and productivity observed for the total of the highly productive teams. CONCLUSIONS: The F&F subfield is in a very dynamic stage, with a great increase in the number of researchers, starting of new teams and consolidation of others. A trend was observed towards the concentration of activity in the most productive teams, that are the most visible internationally both in number of publications in the SCI and in the impact factor of the publication journals. PMID- 9859068 TI - [Oral communication in medicine]. PMID- 9859067 TI - [Systemic mastocytosis. Study of 14 cases]. AB - We have analyzed the clinical, analytical and evolutive data of 14 cases of systemic mastocytosis (SM) diagnosed in our hospital between 1991-1996 and we have compared these results with those of other published series. Clinical parameters, analytical profiles, peripheral hematologic data, radiologic data and histological study were collected. Following Metcalfe's criteria, the patients were classified in 4 groups: a) group 1, indolent mastocytosis; b) group 2, hematologic disorders; c) group 3, aggressive lymphadenopathic mastocytosis with eosinophilia, and d) group 4, mastocytic leukemia. The average age at diagnosis was 52.4 years, range 25-83, and 64% were females. The mean follow-up was 2 yr. In most of the cases (71.4%) the initial complaint was urticaria pigmentosa. The predominant clinical features were pruriginous-eritematous skin lesions (in 11 cases), and digestive symptoms (in 10 patients). The most usual biochemical disorder was the rise of serum alkaline phosphatase level (in 8 patients), while lactate-dehydrogenase (LDH) was normal in all the cases. The most striking roentgenologic features were oteopenia, observed in 50% of our patients. Pheripherical hematological disorders were discovered in 8 patients (64.3%) and in one of them circulating mast-cells were observed. The bone marrow was involved in all patients (100%) and in two of them mielodysplasic features were found. The diagnostic of SM is difficult in the absence of skin lesions. The skin lesions are very common in systemic mastocytosis. Bone marrow involvement is constant, so its study has a high diagnostic rentability. PMID- 9859069 TI - [Cystic fibrosis and digestive involvement: physiopathological, clinical and therapeutical considerations]. PMID- 9859070 TI - [Critical writing of scientific bibliography]. PMID- 9859071 TI - [Toxicity of ecstasy]. PMID- 9859072 TI - [Ecstasy: idiosyncrasy, overdosage or collateral effects]. PMID- 9859073 TI - [Secondary prevention of ischemic stroke: aspirin or ticlopidine]. PMID- 9859074 TI - [Sensitization to kiwi skin]. PMID- 9859075 TI - [Persistent hiccup? Dysfunction of the inferior olivary complex]. PMID- 9859076 TI - [Cefixime versus amoxicillin plus netilmicin in the treatment of community acquired non-complicated acute pyelonephritis]. AB - BACKGROUND: Community-acquired non-complicated acute pyelonephritis (APN) is a frequent, occasionally serious infection (around 20% of the cases are bacteremic) that usually requires hospital admission. The third generation oral cephalosporins which are active against more than 95% of E. coli strains should allow the outpatient management of these patients. OBJECTIVE: To evaluate the bacteriological and clinical efficacy of oral cefixime in comparison to amoxicilin plus netilcilin in the treatment of APN. PATIENTS AND METHODS: Patients older than 18 years affected by APN were included in a fourteen month prospective study. According to a random numbers chart, the patients received cefixime (400 mg/24 h in a single daily dose for 12 days) or amoxicilin (1 g/8 h per os) plus netilmicin (4 mg/kg/24 h in a single intramuscular daily dose) during five days followed by 7 days of an oral treatment chosen according to the susceptibility pattern of isolated microorganism. RESULTS: Sixty-one patients received cefixime and 65 amoxicillin plus retilmicin. There were no significant differences between both groups of patients. Thirty-two patients presented bacteremia (25.4%). The mean (SD) eak and trough concentrations of netilmicin were 11.4 (2.8) mg/l and 0.38 (0.4) mg/l, respectively. Clinical response was favorable in 97% of patients treated with cefixime and in 98% of those treated with amoxicilin plus netilmicin (p = NS). The infection recurred in 10 out of 59 patients (16.9%) in the cefixime arm of the study and in 9 out of 64 patients (14%) treated with amoxicillin plus netilmicin (p = NS). Tolerance to the study drugs was good in both arms of the study, and renal function remained normal. CONCLUSION: Cefixime seems to be an acceptable alternative to the regimens containing an aminopenicillin and an aminoglycoside for the treatment of community-acquired non-complicated APN. PMID- 9859078 TI - [Bibliometric indicators, subjects analysis and methodology of research published in Spain on epidemiology and public health care (1988-1992)]. AB - BACKGROUND: To describe Spanish scientific output in epidemiology and public health through the bibliometric indexes utilization and with regard to subjects, design and statistical methods employed, as well as its appropriateness, importance and funding. MATERIAL AND METHODS: Descriptive, longitudinal and retrospective study over 594 originals published in Spanish medical journals and listed in the Spanish Medical Index (Indice Medico Espanol) during the 5 years period 1988-1992. RESULTS: The average autors by article was of 4.9 and the Lotka's index, for the authors that publish articles on the topic, was -2.92; 82% of the authors have a productivity index equal to zero. Almost 70% of the articles are signed by an isolated institution, being a hospital in 51% of the cases. The journal that publishes most articles on the topic is Medicina Clinica. The average references by article is about 24. The Price' index is 36.5% and the isolation' index is 36.2%. "The non AIDS infections", the descriptive designs and the lack of inferential statistics methods, characterize the content of the articles. Regarding to the criteria established by the study, more than half of the analyzed research (56.5%) was appropriate and almost a fourth (22%) could be considered that was about important topics. CONCLUSIONS: Concerning bibliometric indicators, research on epidemiology and public health in Spain does not seem to differ from other medical specialties. The epidemiology as scientific method is widely used over the medical specialties, something which could explain the wide dispersion of scientific literature on this topic. There seems to be a certain lack of agreement between the research topics and the real health problems of Spain. PMID- 9859077 TI - [Laparoscopic splenectomy as an alternative to open surgery in the treatment of autoimmune thrombocytopenia]. AB - BACKGROUND: Several studies have shown the potential advantages laparoscopic splenectomy (LS) over open surgery. The aim of this study has been to evaluate the advantages of LS over open surgery in the treatment of autoimmune thrombocytopenia. PATIENTS AND METHODS: 54 consecutive patients splenectomized for the treatment of idiopathic thrombocytopenic purpura (ITP) or HIV-related thrombocytopenia were analyzed. Operative features (operative time, conversion to open surgery, accessory spleens), immediate (stay, analgesia and blood transfusion requirements) and late postoperative features (platelet count), as well as splenectomy-related complications in both surgical procedures were compared. RESULTS: Between February 1990 and February 1997, 54 splenctomies were performed for the treatment of autoimmune thrombocytopenia (ITP, n = 47, and HIV related thrombocytopenia, n = 7). Eighteen were performed through an open approach, and 36 by laparoscopy. Both groups were comparable with regard to age, sex, platelet count, disease duration and body mass index. LS was completed in 34 cases (conversion to open surgery: 5.5%). The incidence of accessory spleens was 11% in the LS group and 5.5% in the open surgery group. Postoperative morbidity (16% vs 28%) and blood requirements (25% vs 33%) were lower after LS, but the differences did not reach statistical significance. Analgesia requirements (7 [SD 3] vs 11 [6]; p < 0.01) and postoperative stay (3.8 [2.6] vs 7.4 [3] days; p < 0.01) were significantly shorter after LS. Following splenectomy, the platelet counts became normal in 72% of patients submitted to LS and 78% of patients in the open surgery group. After 20 and 63 months mean follow-up, one patient in each group developed late complications. CONCLUSION: As compared to open surgery, LS offers a better immediate clinical outcome, with similar long-term results. PMID- 9859080 TI - [Evidence based medicine, but in what evidence?]. PMID- 9859079 TI - [Usefulness of anti-neutrophil cytoplasmic antibodies, anti-proteinase 3 and anti myeloperoxidase in management of small vessel vasculitis]. AB - BACKGROUND: Analysis of usefulness of antineutrophil cytoplasmic antibodies (AN CA) as a marker of clinical activity in small vessel vasculitis. PATIENTS AND METHODS: 33 patients, 10 patients with Wegener's granulomatosis (WG) and 23 with microscopic polyangiitis (MPA) and rapidly progressive glomerulonephritis type III (RPGN III). The clinic and serologic follow-up was accomplished every 3 months during an average of 19 (SD, 24) months (range 3-52 months. The serologic follow-up included the determination of ANCA by indirect immunofluorescence (IFI) and ELISA, as well as the serum level of C reactive protein (RCP). RESULTS: At the time of diagnosis all patients were ANCA positive by IFI and ELISA. The seroconversion of ANCA from positive to negative was produced in 30/33 patients (90%). Twenty-six out of these 30 patients (87%) achieved the seroconversion within the first 6 months. During the follow-up 4 patients had a major relapse, all with positive ANCA. In 2 patients, one of each group, seroconversion from negative to positive was not associated with clinical relapse of vasculitis. CONCLUSION: ANCA should be used in conjunction with other indices of disease activity in patients with small vessel vasculitis. PMID- 9859081 TI - [Therapeutic use of growth hormone in the Spanish National Health System]. PMID- 9859082 TI - [Interaction between genes and diet as a determinant of the plasma levels of cholesterol]. PMID- 9859083 TI - [Evidence based medicine: a challenge for the 21st century]. PMID- 9859084 TI - [Placebos in drug clinical trials]. PMID- 9859085 TI - [Diagnosis of the localization of diseased glands in primary hyperparathyroidism: comparison of gammagraphy with 99mTc-sestamibi, echography and computerized tomography]. PMID- 9859086 TI - [Treatment of primary central nervous system lymphoma in AIDS patients]. PMID- 9859087 TI - [The impact of bronchial colonization in the quality of life of patients with chronic, stable bronchitis]. AB - BACKGROUND: The aim of this study was to determine the impact of respiratory function and bacterial colonization of the lower airway on the quality of life of patients with chronic, stable bronchitis (CB). MATERIALS AND METHODS: A series of 41 patients with stable CB was studied (age: 63.8; standard deviation (SD) 9.1 years; FVC% 91.0 (18.9); FEV1% 74.6 (23.7); FEV1/FEC 62.8 (11.2) with normal thoracic radiography. Patients with previous diagnosis of bronchiectasia, bronchial asthma and/or positive bronchodilatory tests (> 15%) were not included in the study. Bacterial growth in a sputum sample of grade 4-5 of the Murray Washington scale was considered diagnostic of bronchial colonization. Measurement of the quality of life was performed with the Nottingham Health Profile (NHP) and the St. George Respiratory Questionnaire (SGRQ). RESULTS: The patients presented a moderate alteration in their quality of life with scores over 25 in most of the dimensions of the NHP and the SGRQ. In 9 out of 41 cases (22%), the sputum cultures demonstrated bronchial colonization with the most frequently isolated bacterias being Haemophilus influenzae and Moraxella catarrhalis. Multivariate analysis performed with the quality of life as the dependent variable showed an association between FEV1/FEC1 and the SGRQ score (R2 = 0.18), and energy (R2 = 0.09) and physical mobility (R2 = 0.05) of NHP. CONCLUSIONS: Bronchial obstruction is the main determinant in the quality of life in patients with stable CB. Colonization of the lower airway is observed in 22% of the patients and also influences the quality of life of the patients but to a much lesser extent. PMID- 9859088 TI - [Relationship between medical treatment compliance and the degree of control in patients with high blood pressure, non-insulin dependent diabetes mellitus and dyslipidemia]. AB - BACKGROUND: To study the relationship between therapeutic compliance and the control of arterial hypertension, non insulin dependent diabetes mellitus and hyperlipidemia. PATIENTS AND METHODS: Prospective study performed on 174 hypertensive patients, 107 with diabetes and 107 with hyperlipidemia evaluating compliance by counting of tablets in two home visits. RESULTS: 34% hypertensive patients, 20% diabetics and 37% hyperlipidemics that took medication as instructed or more than they should were badly controlled. CONCLUSIONS: The control grade of high blood pressure, non insulin dependent diabetes mellitus and hyperlipidemia not only depends on improving compliance but also in adapting pharmacologic prescriptions. PMID- 9859089 TI - [Medication use in diabetes mellitus (VI). Economics and effectiveness of insulin and sulfonyl-urea combination therapy compared with conventional two daily doses]. AB - BACKGROUND: To compare the cost and effectiveness of bedtime intermediate-acting insulin and daytime-sulfonylurea (SU) combination therapy versus the conventional two-daily-dose insulin treatment. SUBJECTS, MATERIAL AND METHODS: A pharmacoeconomical analysis of cost minimization. To prove a similar effectiveness a transversal prospective study was carried out. Patients recently converted to insulin due to oral hypoglycaemic agents failure were recruited. Entry criteria were: age > 40 years-old, more than 3 and 1 years of diagnosed diabetes and follow-up, respectively, current BMI between 20-40 kg/m2, baseline HbA1c > 8.5% and fasting C-peptide > 0.3 nmol/l. BMI, HbA1c, hypoglycaemic crisis, insulin and SU (glicazide and glibenclamide) daily dose were recorded, estimating the cost of both therapies. RESULTS: Sixty-five patients (23 male), 32 in combined therapy (mean daily dose of insulin 19.5 U and 2.4 SU tb t.i.d.) and 33 patients with a two-insulin-injection regimen (38.4 U) were treated during a follow-up period of 2.4 years. The two groups exhibited similar mean age (67.8/67.7y), known diabetes duration (15.9/15.1y), BMI (28.9/28.8/kg/m2), previous HbA1c (8.9/9.1%) and fasting C-peptide (1.6/1.2 nmol/l). No statistical differences in BMI increase (1/1.4 kg/m2), neither in mean HbA1c (7.8/7.9%) nor severe hypoglycaemic crisis (0.03/0.17 episodes/year) were evidenced. Patients in combined therapy reported a lower number of mild hypoglycaemic crisis (0.7/1.9 episodes/month; p < 0.01) and the daily cost was significantly lower (94.5/134.3 ptas./day; p < 0.0001). CONCLUSIONS: Both therapies, two-insulin-injection regimen and insulin and sulfonylurea combination therapy were similarly effective in having an acceptable glycaemic control with similar risk for weight gain or severe hypoglycaemia. Combined therapy was more cost-effective and well tolerated, thus, comfort and a lower risk of mild hypoglycaemic episodes were evidenced. PMID- 9859090 TI - [European study of asthma. Prevalence of atopy in young adults of 5 areas in Spain. Spanish Group of European Asthma Study]. AB - BACKGROUND: The aim of the current study is to show the prevalence of atopy in five Spanish areas, and its variability according to area, age and gender. PATIENTS AND METHODS: From a populational based sample of 16,884 individuals aged 20 to 44 years-old, we obtained a randomized 20% subsample (n = 3,310). Participants performed specific IgE measurements, skin prick tests, forced spirometries and metacholine challenges to measure bronchial hyperresponsiveness. The response rate was 40%, and 1,313 individuals were finally included in the study. Specific atopy to the following aeroallargens was determined: cat dander, Cladosporium, Dermatophagoides, Phleum, Parietaria, birch, Alternaria, ambrosia, olive, rye grass and dog dander. RESULTS: The global prevalence of atopy (detectable specific antibodies IgE in serum and/or skin reactivity) widely varied by area, skin reactivity ranking in males from a minimum in Albacete (24.6%; 95% CI: 18-33) to a maximum in Huelva (39.6%; 95% CI: 30-53), and in females ranking from a minimun in Galdakao (10.3%; 95% CI: 6-17) to a maximum in Barcelona (28.8%; 95% CI: 19-43). Considering separately seropositivity and skin reactivity we observed a similar trend. Males showed a higher prevalence of global atopy (40.1%) than females (29.4%). Our data indicate that there is a decrease in the prevalence of atopy according to age in the general population, but only significant in men. Dermatophagoides pteronyssinus is the most common allergen in all ares but Albacete, where the most common allergen is the olive pollen. CONCLUSIONS: By means of a standard methodology, we report population data of the prevalence of atopy in five Spanish areas. The distribution of the prevalence of atopy varies widely in the five areas surveyed, according to the composition of the most common environmental allergens. PMID- 9859092 TI - [Chronic obstructive airways disease and perceived health]. PMID- 9859091 TI - [Markers of hemostatic activation in the coronary sinus versus peripheral circulation in coronary artery disease. Effects of percutaneous transluminal coronary angioplasty]. AB - BACKGROUND: To evaluate haemostatic changes in coronary sinus (CS) and peripheral circulation (PC) in patients with cardiovascular disease (CAD) following PTCA. PATIENTS AND METHODS: We studied 12 patients scheduled for elective PTCA. Blood samples were obtained from CS and PC before angioplasty and after each balloon dilation. Expression of P selectin and several haemostatic parameters were determined. RESULTS: D-Dimers (DD) plasma levels were significantly increased in samples from CS. No significant differences were found in other haemostatic parameters. A significant decrease of thrombin-antithrombin III levels after PTCA was assessed. CONCLUSIONS: Intracoronary and peripheral hemostatic activation does not occur during and immediately following PTCA. No major differences in hemostatic parameters between CS and PC were observed in patients with CAD. PMID- 9859094 TI - [Ethical review of epidemiologic studies: a need and a proposal]. PMID- 9859093 TI - [Dengue: a re-emerging disease. A clinical and epidemiological study in 57 Spanish travelers]. AB - Dengue infection is nowadays considered a re-emergent disease. It has a worldwide tropical and subtropical distribution. The dengue virus in a member of the flavivirus family composed by 4 different serotypes. The virus is transmitted by mosquitos of the Aedes genus. With the increment of travels to the endemic areas, dengue is now observed frequently in our country. We analyzed 57 patients, 30 with imported dengue (ID) and 27 with dengue fever suffered during the trip (DDT). This series is compared with other published ones and a review of the subject is presented. Patients with ID followed a protocol as a febril syndrome returning from the tropics. Dengue was diagnosed through a compatible clinico epidemiological history, the absence of other ferbil illness and positivity of specific serology. All patients had travelled to endemic areas (Central America 28 cases, Indian subcontinent 15, South-East Asia 10, South America 2, West Africa one, and Pacific one). The following were the most important clinical characteristics: fever and asthenia (100%), headache (98%), mialgia (84%), arthralgia (72%), morbilliform rash (61%) and retroocular pain (65%). For ID cases, the most helpful analitical results were: leucopenia (70%), reactive lymphocytes in peripheral blood smear (70%), thrombocytopenia (70%), and increased hepatic enzymes ALAT (53%), ASAT (63%) and LDH (100% in the 7 patients tested for this enzyme). Dengue must be included in differential diagnosis of fever in patients coming back to travels to tropical areas. PMID- 9859095 TI - [Regulatory proteins of the cell cycle: alterations in the cycline D1 pathway as a paradigm. Findings in breast cancer]. PMID- 9859096 TI - [Evidence-based medicine and anti-hypertensive treatments]. PMID- 9859097 TI - [On "The national tuberculosis control program for Spain"]. PMID- 9859098 TI - [Protease inhibitor-associated hypertriglyceridemia and diabetes mellitus in HIV infected patients]. PMID- 9859099 TI - Cardiopulmonary resuscitation in Saudi Arabia. A proposal for the formation of a Pan-Arab Resuscitation Council. PMID- 9859100 TI - Advisory statements of the International Liaison Committee on Resuscitation (ILCOR) CPR in pregnancy. PMID- 9859101 TI - Initial management of patients in trauma and shock. PMID- 9859102 TI - Pulse oximetry: an added criterion for discharge from the post-operative care unit. AB - Based on the observation that the degree of wakefullness measured by Post Anesthetic Recovery (PAR) score in some patients does not correlate with their oxygen saturation, the authors decided to carry on a study to validate that assumption. Three hundred patient ASA I & II were studied. Oxygen saturation and PAR score were recorded from the time of arrival till their discharge by the recovery room staff nurses. Thirty one patients out of one three hundred (10%) were found to be hypoxic (saturation < 95%) despite their high PAR score. The degree of wakefullness as measured by PAR scores cannot be used to establish an end point for oxygen supplementation. Oxygen supplementation and SpO2 monitoring are recommended in all patients recovering from anesthesia. PMID- 9859103 TI - Critical incident reports. AB - We describe a retrospective analysis of critical incident reports in two teaching hospitals. We included significant observations, involving unsafe practices during cardio-pulmonary resuscitation intensive care management and during anesthesia. Of the 143 critical incidents reported, 87% did not lead to negative out-come, out of these 13% were reports on deaths of patient resuscitated by CPR team or emergency department, underwent surgery, and or managed in the intensive care unit. Human errors and lack of communications were common factors for the majority of the incidents. Wrong drug labeling and irresponsible behavior were the most frequent among the human errors. The analysis aimed to regularize the method of reporting and also to determine the causes of complications, offer solutions and prevent occurrence of such incidents in the future. PMID- 9859104 TI - Anesthesia for laser surgery. PMID- 9859105 TI - Loss of vision: a manifestation of TURP syndrome. A case report. AB - We report a case of transient blindness, part of a TURP syndrome, in a 65 year old man undergoing TUR of the prostate under centro-neuron block. Irrigating fluid consisted of 1.5% glycine. As a result, serum levels of glycine increased to 12,470 mols.l-1 (N176-332 mumols.l-1). We attribute this loss of vision to glycine toxicity. PMID- 9859106 TI - Evoked potentials in high cervical spine injury. PMID- 9859107 TI - HELLP syndrome: undiagnosed in a case of impending rupture of uterus. A case report. AB - A 31-year-old women, 34 weeks gestation with intrauterine fetal death, and transverse lie with impending rupture of the uterus due to obstructed labor, was scheduled for urgent cesarean section. On preoperative anesthetic assessment, she was diagnosed to have HELLP syndrome based on clinical findings (subsequently confirmed by laboratory results). She was anesthetized taking the necessary precautions. Her intra and postoperative course is described. The case demonstrates how the anesthetist is often confronted by undiagnosed HELLP syndrome for urgent cesarean section and has limited time for investigations. He should depend on his clinical acumen to diagnose and treat appropriately for a favorable maternal outcome. PMID- 9859108 TI - Reprogramming the phagocytic pathway--intracellular pathogens and their vacuoles (review). AB - Phagocytic immune cells (particularly macrophages and neutrophils) take up and digest particles that have invaded our bodies. In doing so, they represent a very early line of defence against a microbial attack. During uptake, the particles are wrapped by a portion of the phagocyte's plasma membrane, and a new endocytic compartment, the phagosome, is formed. The typical fate of a phagosome is its fusion with lysosomes to yield a phagolysosome in which the particle is digested. Recent data show that some 'intracellular microorganisms' that can cause severe illnesses (tuberculosis, leprosy, legionnaire's disease and others) manage to reprogramme the host phagocytes not to deliver them to the lysosomal compartment. This probably results in increased survival of the pathogens. The analysis of the composition of such 'novel' compartments and research on the molecular mechanisms underlying the microbial interference with host cell functions are likely to yield important insights into: (1) which endocytic/phagocytic compartments phagocytes employ to handle ingested material in general; (2) how some pathogenic microorganisms can reprogramme the phagocytic pathway; and possibly (3) how infections caused by these microorganisms can be treated more effectively. Here, some studies are presented analysing which compartments intracellular pathogens inhabit and how microbes might be able to reprogramme their host cells. PMID- 9859109 TI - Phosphatidylinositol 4-kinase of Torpedo californica electrocytes: physico chemical characterization and regulation by calcium and vicinal molecules of phosphatidylinositol. AB - A phosphatidylinositol 4-kinase (Ptdlns 4-kinase, M(r) approximately 95,000) from the membranes of the electric organ of Torpedo californica was purified to apparent homogeneity. The Michaelis constant for ATP (KM = 280 +/- 60 microM at 20 degrees C) and the inhibition constant for adenosine (Ki = 0.4 mM at 20 degrees C) qualify the electrocyte Ptdlns 4-kinase as a type III kinase. The Ptdlns 4-kinase phosphorylates preferentially exogenous Ptdlns, added in the form of mixed Ptdlns/Triton X-100 micelles, whereas endogenously bound Ptdlns in the membrane fragments of electrocytes is a very poor substrate. It is important that the enzyme and the substrate Ptdlns are situated in different lipid bilayers. The catalytic turnover constant for exogenous Ptdlns is k = 55.3 +/- 6 min-1 at 20 degrees C and the molar Triton X-100/Ptdlns ratio of 16:1. For the substrate Ptdlns in the 'micellar solvent' Triton X-100, steady state kinetics were analysed in terms of the mole fraction X = n(Ptdlns)/[n(Ptdlns) + n(Triton X)] yielding the characteristic Michaelis mole fraction XM = 0.019 +/- 0.005 at 20 degrees C. The activity of the enzyme was enhanced about 5-fold in the presence of Triton X-114, yielding k = 277 +/- 30 min-1 at 20 degrees C. Triton X-114 has a shorter head-group, indicating that the vicinity of the Ptdlns head group in the mixed micelles should not be screened by bulky neighbours. The inhibition of the enzyme activity by Ca2+ is highly cooperative yielding the Hill inhibition constant Ki = 0.47 +/- 0.1 mM and the Hill coefficient h = 3.6 +/- 0.5. The enthalpy of activation is 100 +/- 10 kJ/mol between 0 degree C and 20 degrees C. Although the Ptdlns 4-kinase can be affinity-chromatographically copurified with the nicotinic acetylcholine (AcCho) receptor, suggesting tight association between the two proteins. AcCho does not affect the activity of the Ptdlns 4 kinase in the presence of the AcCho receptor. PMID- 9859110 TI - TGN38 cycles via the basolateral membrane of polarized Caco-2 cells. AB - TGN38 is a heavily glycosylated, type I integral membrane protein which is predominantly localized to the trans Golgi network (TGN), but which constitutively traffics between the TGN and the cell surface. The trafficking of TGN38 has been extensively studied in non-polarized cells, and a short, tyrosine based, peptide motif within the cytosolic domain of the protein has been shown to be necessary and sufficient for its rapid internalization from the cell surface and efficient delivery to the TGN. Such tyrosine-based motifs have also been shown to act as basolateral targeting signals, whilst N-linked glycans (as occur on the extracytosolic domain of TGN38) can act as apical targeting signals. TGN38 has previously been shown to be sorted to the basolateral surface of polarized canine MDCK cells; a polarized cell line in which biosynthetic sorting decisions concerning the eventual destination of apical or basolateral targeted plasma membrane proteins are made at the TGN. We now show that TGN38 is targeted exclusively to the basolateral domain of polarized human Caco-2 cells, a cell line in which newly synthesized membrane proteins destined for either the apical or basolateral plasma membrane may be sorted for delivery to their final destination either at the TGN or at the cell surface. These data also demonstrate that the heavily glycosylated, extracytosolic domain of TGN38 does not contain a dominant apical targeting signal. PMID- 9859111 TI - Oligomerization of the cytotoxin alpha-sarcin associated with phospholipid membranes. AB - alpha-Sarcin is a cytotoxic protein that specifically inactivates ribosomes. The protein translocates across phospholipid membranes. Oligomerization of the protein occurs upon interaction with membranes. Chemically cross-linked protein oligomers have been obtained by treatment of protein-vesicle complexes with the membrane impermeant reagent bis-(sulfosuccinimidyl) suberate. These structures are only obtained in the presence of acidic lipid vesicles composed of either natural or synthetic phospholipids. Such oligomers are not produced in concentrated protein solutions in the absence of vesicles. The formation of the chemically stabilized oligomers is saturated at the same lipid to protein molar ratio as all the perturbations caused by alpha-sarcin on lipid vesicles. Results are discussed in terms of the involvement of oligomer formation on protein translocation across membranes. PMID- 9859112 TI - Effect of procaine on membrane potential and intracellular pH in Xenopus laevis oocytes. AB - The effect of the local anaesthetic procaine on the intracellular pH, pHi, and electrophysiological properties of full-grown Xenopus oocytes was studied. In spite of its interference with both the pH-sensitive microelectrodes and fluorescent probe BCECF, we have shown that procaine induced an intracellular acidification rather than the alkalization commonly observed in most cells. The resting pHi of Xenopus oocytes loaded with BCECF was 7.36 +/- 0.04 (n = 16). Addition of 10 mM procaine to the bath at pH 7.5 caused pHi to decrease to a new steady state value of 6.97 +/- 0.05 (n = 9). A similar behaviour of pHi was observed with microelectrodes. Procaine also promoted a rise in membrane conductance and a membrane depolarization. These changes in membrane potential and conductance were not caused by the decrease in pHi since the addition of sodium propionate at pH 7.5 produced the same decrease of pHi as procaine, but resulted in only a slight depolarization with superimposed oscillations. Current measurements using two-electrode voltage clamp showed that the depolarization was associated with an inward current. No significant effect on this current was observed when replacing Cl, K or Na in the external medium. The absence of effect of Cl and K channel inhibitors argues against the involvement of Cl and K currents during the procaine response. PMID- 9859113 TI - Two glycoprotein populations of band 3 dimers are present in human erythrocytes. AB - The human erythrocyte Band 3 anion exchanger contains a single site of N glycosylation that contains either a short complex oligosaccharide or an extended polylactosaminyl oligosaccharide. Approximately equal amounts of the different glycosylated forms of Band 3 are found in human red cells. As Band 3 exists predominantly as dimers, they may be uniform and consist of a subunit containing an extended oligosaccharide paired with a subunit containing a short oligosaccharide chain. Alternatively, Band 3 dimers may be comprised of subunits that either contain polylactosaminyl or short oligosaccharide chains. To distinguish between these two extremes, the ability of Band 3 membrane domain dimers to bind to immobilized tomato lectin, which specifically binds polylactosaminyl oligosaccharide, was tested. The dimeric membrane domain of Band 3 could be resolved into two fractions by tomato lectin chromatography. This shows that Band 3 dimers are not homogeneous and that two pools exist in red cells, some with a long polylactosaminyl oligosaccharide and the other with a short complex type. The amount of short chain form recovered in the unbound fraction was higher than expected for a random distribution of oligosaccharide chains on Band 3 dimers. Detergent extraction experiments showed that Band 3 glycoforms did not display a differential interaction with the cytoskeleton. The ability to separate Band 3 dimers into two glycoform populations suggests that subunit exchange between dimers does not occur in the membrane or in detergent solution. Furthermore, the results show that while one population of Band 3 dimers is processed to contain polylactosaminyl oligosaccharide, the other largely escapes this processing step. PMID- 9859114 TI - [Lithium salts in child and adolescent psychiatry]. AB - Lithium is--besides neuroleptics--the drug of choice for the treatment of manic episodes. If the use of antidepressive drugs, during unipolar depressive illness, does not lead to a positive response, the additional administration of lithium is appropriate, even during a depressive episode. Lithium is also considered as the drug of choice for prophylactic treatment of bipolar affective disorders. This holds true also for adolescents. In contrast to the indications in adults, in adolescents an early administration is desirable to reduce risk factors of psychosocial development. Additional indications may be the presence of severe aggressivity in conduct disordered children. In these cases, a treatment with lithium salts can result in a behavioral improvement. This may be also the case in impulsive self-injurious behavior. The dosage and serum levels of lithium, as well as its adverse effects are comparable with those known from adults. At present, lithium treatment cannot be recommended for children under 12 years of age--except under in-patient conditions. PMID- 9859115 TI - [Significance of hepatic cytochrome P450 enzymes for psychopharmacology]. AB - Nearly all psychotropic drugs are metabolized by hepatic cytochrome P450-enzymes. In humans, there are 5 isoenzymes involved in this process. The activity of these enzymes can be modulated by a number of commonly used drugs, yielding potentially hazardous interactions. Most of the recently introduced selective serotonin reuptake inhibitors are potent inhibitors of cytochrome P450 enzymes. Thus, the plasma concentrations of tricyclic antidepressants or clozapine might be elevated into toxic levels. In contrast, carbamazepine induces most of the isoenzymes. This potentiates the elimination of tricyclics and antipsychotics and might cause a serious risk for the recurrence of depressive or psychotic symptoms. Moreover, 5-10% of the population are slow metabolizers of CYP2D6. This group is prone to increased adverse effects of moderately dosed medication. This review systematically points out the reported or predicted pharmacokinetic drug interactions in psychopharmacology focussing on clinical significance. PMID- 9859116 TI - [Delusion in depression]. AB - The present article endeavours an outline of depressive delusions, their symptomatology and their various topics. Their relations to depressive feelings of guilt and anxiety are discussed and differentiated. Delusions of guilt, poverty or disease including their preliminary states are by no means rare but regularly occurring symptoms which substantiate the diagnosis of major depressive disorder/endogenous depression/melancholia. Preliminary states like hypochondriatic fears of guilt and poverty are likewise characteristic clinical signs which may be differentiated with high validity from the kind of anxiety associated to neurotic depression/dysthmia. Delusional depression did not prove to be a nosological entity but as a concept may bear considerable heuristic value concerning therapeutic considerations. This particularly serious form of depression requires specific therapeutic procedures. PMID- 9859117 TI - [Adjuvant whole body acupuncture in depression. A placebo-controlled study with standardized mianserin therapy]. AB - In order to examine the efficacy of whole body acupuncture additionally applied to drug treatment in depression, a single-blind placebo-controlled study with 70 inpatients administered to three different treatment groups has been carried out. All patients were pharmacologically treated with the tetracyclic antidepressant mianserin. The verum group (n = 22) received acupuncture at specific points considered to be effective in the treatment of depression. The placebo group (n = 24) was treated with acupuncture at non-specific locations and the control group (n = 24) received only pharmacological treatment. Acupuncture was applied three times a week over a period of four weeks. Psychopathology was rated by judges blind to verum/placebo conditions twice a week over eight weeks with the CGI, GAS, BRMS and BfS rating scales. Additionally applied acupuncture improved the course of depression more than pharmacological treatment with mianserin did by itself. However, we could not detect any differences between placebo and verum acupuncture. PMID- 9859118 TI - [Volumetric brain findings in late depression. A study with quantified magnetic resonance tomography]. AB - A number of observations including clinical manifestation, course, outcome, and family history, support the view that patients presenting with a major depression occurring first in late life should be treated as a nosological subgroup. In this study quantitative magnetic resonance imaging (MRI) was used to investigate volumes of different brain structures in 19 patients with late onset major depression (age of onset > 50) and 13 age matched controls. 3-D MRI sequences were acquired using a Siemens 1.5T scanner. Whole brain volume, CSF volume, volume of the frontal and temporal lobes and the volume of the amygdala hippocampus complex were assessed using the software NMRWin. Compared to the controls, depressed patients showed a significantly lower whole brain volume and a significantly higher CSF volume, whereas volumes of the frontal and temporal lobes as well as the amygdala-hippocampus complex volumes were not significantly decreased. In addition, depressed patients exhibited a higher ventricle-brain ratio suggesting a higher degree of central atrophy compared to healthy individuals. Our results indicate that cerebral changes involving subcortical structures are of relevance in the pathogenesis of late-onset depression. Defining the aetiology of these lesions may be important for the development of preventive treatment of depression in the elderly. PMID- 9859119 TI - [Mild cognitive deficit in the elderly. Results of a gerontologic study]. AB - The term "mild cognitive impairment" refers to cognitive deficits which exceed normal physiological aging processes, but do not fulfill the criteria for dementia. The prevalence rates of four current concepts were compared in a sample of 202 healthy 60-64 year-old participants recruited from the interdisciplinary longitudinal study on adult development and aging (ILSE). Furthermore, the relationships between cognitive deficits and psychological and sociodemographic variables were examined. The following prevalence rates were determined: 13.5% for age-associated memory impairment, 6.5% for age-consistent memory impairment, 1.5% for late-life forgetfulness and 23.5% for aging-associated cognitive decline. Subjective cognitive complaints did not correlate with results obtained from neuropsychological tests. Significant correlations were however found between subjective cognitive complaints and higher scores on depression and neuroticism scales. Significant correlations were also found between a reduced test performance and a lower educational level and socioeconomic status. Longitudinal studies are warranted to further elucidate the predictive value of these diagnostic concepts. PMID- 9859120 TI - [Dementia screening in routine clinical practice. A comparative analysis of MMSE, SIDAM and ADAS]. AB - Dementia-screening in clinical routine requires short, sensitive and specific tools. A number of standardized instruments are available for this purpose. The present study analysed the relationship between size of three exemplary dementia screening tests and their diagnostic accuracy. The Mini-Mental-State-Examination (MMSE), the Structured Interview for the Diagnosis of Dementia of the Alzheimer type, Multiinfarct Dementia and Dementias of other Aetiologies according to ICD 10 and DSM-III-R (SIDAM) and the Alzheimer's Disease Assessment Scale (ADAS) were applied to 71 patients with dementia of the Alzheimer-type and 73 non-demented controls. A ROC-analysis revealed that neither SIDAM nor ADAS differentiated better between demented and non-demented probands than the MMSE. This was also true for patients with mild dementia. In dementia staging the more comprehensive instruments did not surpass the MMSE, too. Due to it's brevity, the MMSE is the preferential screening-instrument for clinical routine. PMID- 9859121 TI - [Copying and free drawing by patients with Alzheimer disease of different dementia stages]. AB - In this study we assessed the drawing abilities in 37 patients with probable Alzheimer's disease according to NINCDS-ADRDA criteria. Drawing abilities (drawing: house, flower, clock; Rey-Osterrieth figure; copying: MMST-figure; Rey Osterrieth figure) were quantified with different rating schemes and related to other neuropsychological assessments. All patients underwent a positron emission tomography with 18-FDG. Drawing performance was highly correlated with severity of dementia--expressed in MMST scores (r = 0.78; p < 0.0001)--with visuo-spatial short-term memory (r = -0.69; p = 0.001), and writing abilities (r = -0.77; p < 0.0001). The summarized drawing score showed a statistically significant correlation with the rate of temporoparietal glucose metabolism measured with positron emission tomography and 18-FDG (r = 0.39; p = 0.017). In the drawings of AD patients omittings and simplifications were typical, whereas perseverations rarely occurred. In severely demented patients closing-in phenomenons could be described, too. A subgroup of AD patients with visuo-constructive impairment as the leading symptom could not be identified. PMID- 9859122 TI - [Prescribing neuroleptics to senile dementia patients. On outcome in old age homes after inpatient psychiatric treatment]. AB - Psychopharmacological treatment of all demented patients (n = 49), discharged to nursing homes after gerontopsychiatric hospital treatment in 1992 and 1993, was analyzed. In spite of the higher risk of secondary effects in elderly, demented people are after prescriptions for psychopharmacologic medications, especially neuroleptics. It was shown that the residents in nursing homes receive significantly more neuroleptics than patients who are discharged (P = 0.0001). Prescriptions for benzodiazepines were two times higher, whereas clomethiazole was rarely given. In nursing homes these substances were prescribed as continuous treatment. Most prescriptions resulted from agitation, sleep disturbances or aggressive behavior. Astonishingly, the patients without positive effects from treatment, who worsened regarding behavioral complications of dementia, obtained the highest doses of neuroleptics. PMID- 9859123 TI - [Kaufman Assessment Battery for Children. Differential evaluation of (partial) intellectual ability of mentally handicapped adults]. AB - The use of the Kaufman-Assessment Battery for Children (K-ABC) was evaluated with a sample of 50 mentally disabled adults in a wide age range, as there exist no special tests for this group of patients. Intercorrelation--and factor analyses, and correlations with the German adaptation of the Wechsler Adult Intelligence Scale (HAWIE-R) proved the use of the K-ABC to be valid. As an assessment battery for children designed for the wide age range from 2-6 to 12-5 years is more adequate to the capability of intellectually impaired adults, the K-ABC delivers results on individual strengths and weaknesses, based on a neuropsychological theory. Thus the individual cognitive abilities can be assessed with more differentiation, and that gives an important base for intervention, partial remediation, and placement. PMID- 9859124 TI - [Delirium during oral therapy of herpes zoster with acyclovir. Case report and brief review of central nervous system side-effects of acyclovir]. AB - In differential diagnosis of a delir also adverse effects of medicaments have to be taken into account beside other causes. We report a case of an agitated delir with nocturnal disturbance of consciousness, confusion, restlessness and sleeplessness. This delir existed exclusively during the therapy of a cutaneous herpes zoster with zovirax-pills which can only be explained by a causal connection--after exclusion of other causes. As a so far undescribed predisposition for neurotoxicity of oral therapy with acyclovir signs of vascular encephalopathy were found in the patient's cranial magnetic resonance imaging. The central nervous side effects of acyclovir were summarized shortly. PMID- 9859125 TI - [High dosage thyroxine treatment in therapy and prevention refractory patients with affective psychoses]. AB - The following review summarizes current knowledge on the treatment of therapy resistant patients with affective disorders with supraphysiological doses of thyroxine (T4). Several groups have reported independently of each other that administration of 200-500 micrograms T4/day has excellent effects in 50-65% of patients a) with bipolar disorder, with or without, "rapid cycling" course, who were previously resistant to all prophylactic drugs and b) in the treatment of therapy-resistant depression. T4 is effective only in combination with an antidepressant or a prophylactic drug. Side effects are minimal, even when T4 is administered over several years. These results now justify to recommend high dose T4-augmentation as "last-resort" treatment also beyond research purposes, i.e. in psychiatric wards and in private practice. Recommendations for clinical applications are given and hypotheses on possible mechanisms underlying the efficacy of T4 treatment are discussed. PMID- 9859126 TI - [Propagation of the pathogen of Lyme disease Borrelia burgdorferi in the ticks Ixodes persulcatus]. AB - An increase of the number of Borrelia burgdorferi s. l. in nymphs was observed 3 4 months later the moulting and during the following 4-5 months. The percent of infected ticks remained unchanged. The reproduction of spirochete was going not only at a room temperature, but also at a lower temperature, 3-5 degrees (imitation of hibernation conditions). An increase of the spirochete density was especially well marked, when the ticks were maintained under fluctuated daily conditions. We have found out the difference in the transphase transmission. Almost all larvae and nymphs infected by a bloodsucking retained spirochetes after moulting (the first transphase transmission). When the nymphs infected at larval stage were fed on an uninfected mouse, only 30% of imago ticks retained the spirochetes (the second transphase transmission). PMID- 9859127 TI - [Features of the parasitic system of Ixodid ticks--Borrelia--small mammals in the Russian Northwest]. AB - Mammals and ticks was collected in 1995-1997 years in Novgorod region from May to September. Traps were exposed during 5 consecutive days every month in 4 different biotopes. 928 hosts were captured during three years. 357 larvae and 59 nymphs of Ixodes persulcatus, 356 larvae and 71 nymphs of I. trianguliceps were collected. The Clethrionomys glareolus and Sorex araneus are main hosts of larvae of I. persulcatus and I. trianguliceps. C. glareolus feed 76.2% of I. persulcatus larvae and 25% of I. trianguliceps larvae, S. araneus--27.1 and 45.9% of larvae, respectively. The abundance of larvae and nymphs of I. persulcatus was 0.26 and 0.13 on C. glareolus, and 0.41, 0.08 on S. araneus; for I. trianguliceps this indices are 0.15 and 0.09, 0.95 and 0.13, respectively. Infection rate of larvae and nymphs I. persulcatus and I. trianguliceps with Borrelia burgdorferi s. l. was 94-100%, confirmed by IFA with H5332. The borreliemia was found in 13.6% G. glareolus, 6.6--Microtus arvialis, 4.2% Apodemus uralensis. For all species of sorex this index varied from 0.9 to 44.9% in different years. Increase of mammal's numbers with the borreliemia correlated with peaks of activity of larvae of I. persulcatus. Finding of Borrelia burgdorferi s. l. in I. trianguliceps indicates an occurrence of enzootic locuses. PMID- 9859128 TI - [Isocitrate dehydrogenases of trematodes parasitizing cattle and the feasibility of inhibiting them using anthelmintic preparations]. AB - Activities and properties of NADF-dependent isocitrate dehydrogenases in cytosol and mitochondrial fractions from trematodes Eurytrema pancreaticum and Calicophoron ijimai were examined. Cytosol and mitochondrial enzymes were activated by ions Mn2+ and Mg2+ and inhibited by ions of heavy metals and p chloromercuribenzoate. The effect of anthelmintic preparations on activity of enzymes was investigated. PMID- 9859129 TI - [Names of scientists in valid names of mosquitoes (Diptera: Culicidae), initially described from the territory of the former USSR]. AB - The paper gives brief biographic data biologists, to whom the valid names of mosquitos, described from the former USSR, had been dedicated. PMID- 9859130 TI - Where is the person in the case? PMID- 9859131 TI - Slow rehabilitation of a traumatic lower limb amputee. AB - A 65-year-old male (GM) was referred to the physiotherapy department of the regional limb fitting centre for outpatient rehabilitation following a right transfemoral amputation five weeks previously. He had been knocked down by a bus and suffered a crush injury to the right leg resulting in a stable fracture to the right pubic ramus, a fractured skull and orbital bone. Immediately following admission to the local district general hospital, his right leg was amputated at the transfemoral level. He was nursed post-operatively in ITU for three days and was ventilated during this time. GM was then transferred to the general orthopaedic ward. He received physiotherapy throughout his hospital stay by ward based physiotherapists--not experts in amputee management but with access to specialist advice. Early physiotherapy was primarily concerned with respiratory care and maintenance of limb mobility and function in bed. Gentle, active stump exercises were commenced on the first post-operative day. GM sat out of bed on the sixth day and stood with the support of two people on the eighth day. His wound was healing well and treatment in the physiotherapy gym began the same day. Use of the pneumatic post-amputation mobility aid (PPAM aid) (Redhead, 1983; Marks, 1996) was started on day nine. However, progress with this early walking aid (EWA) was slow and GM achieved independent walking using parallel bars on the eighteenth post-operative day. PMID- 9859132 TI - Rehabilitation of a traumatic lower limb amputee. PMID- 9859133 TI - The accuracy of predicting functional recovery in patients following a stroke, by physiotherapists and patients. AB - BACKGROUND AND PURPOSE: The potential for post-stroke recovery and the range of predictive variables has been studied extensively. Knowledge of these variables alongside other factors, such as performance in therapy and professional experience, enable ongoing predictions to be made by members of the rehabilitation team. Patients' own predictions for their recovery has yet to receive much attention in this area of research. The aim of this study was to compare the predictive accuracy of the physiotherapist and the stroke patient with regard to functional change during a period 6-12 weeks post-stroke. METHOD: The stroke sample (N = 29) came from two National Health Service Trusts as did the physiotherapists (N = 4). No comparisons were made between the hospitals and data was coded for anonymity. Estimations were made by both physiotherapists and patients regarding items on each of the three sections of The Rivermead Motor Assessment (RMA). Intra-class correlation coefficients (ICCs) were used to describe agreement of each set of predictions with the achieved RMA scores. The results reported here represent the main emphasis of the research; however, other areas were also screened (for example, change in cognition, language and quality of life) by use of basic standardized measures. Recovery was also compared to other known predictive variables, such as age, severity of stroke and urinary incontinence. RESULTS: At follow-up assessment it was found that both physiotherapists' and patients' predictions demonstrated high and significant agreement with the achieved RMA scores at 12 weeks (ICCs ranging from 0.727 to 0.968). Physiotherapists' predictions demonstrated marginally higher levels of agreement than patients' predictions. CONCLUSIONS: The degree of accuracy demonstrated by both physiotherapists and patients was considerable. The patient group was perhaps the more notable as no subject had had prior knowledge of a stroke. The implications in respect of lay persons' involvement in decision making and in the rehabilitative process, alongside the health professionals, are perhaps worthy of closer consideration. PMID- 9859134 TI - Learning physiotherapy: students' ways of experiencing the patient encounter. AB - BACKGROUND AND PURPOSE: The aim of this paper was to describe and analyse the impact of formal education and professional experience on physiotherapy students' ways of experiencing interaction within a patient encounter. METHOD: Two groups of physiotherapy students were interviewed on two occasions; during the second and last term of their formal education programme, and during the last term and after 18 months' professional experience. Data were subjected to a qualitative analysis. Changes in conceptions between the two interview occasions were described quantitatively. RESULTS: Subjects' ways of experiencing interaction within a patient encounter could be described in four main categories: Mutuality; Technicalism; Authority and Juxtaposition. Mutuality and Technicalism denoted an integration of the communicative and problem-solving processes involved in the encounter, the former category from a patient-centred and the latter from a physiotherapist-centred perspective. Authority and Juxtaposition denoted a separation of the processes, the former from a physiotherapist-centred perspective and the latter from a patient-centred one. CONCLUSIONS: The results show a trend as regards direction of change in conceptions from separated to integrated perspectives on the communicative and problem-solving processes after the formal education programme. After 18 months' professional practice the Mutuality category dominated. PMID- 9859135 TI - Reliability of clinical balance outcome measures in the elderly. AB - BACKGROUND AND PURPOSE: Simple, practical and reliable clinical balance outcome measures are needed to assess baseline status and response to treatment in older people. The reliability of the clinical measures used in this testing protocol had not been determined for this population. This study assessed the inter-rater reliability of three commonly used clinical measures of balance: one leg standing, tandem gait and functional reach. METHOD: Two samples of older people were used: (1) non-disabled and (2) disabled community dwellers. All testing was performed in a single session by two trained examiners according to a standardized protocol. Intra-class correlations were calculated comparing the means of each clinical balance test for Examiner 1 with Examiner 2. RESULTS: Reliability coefficients were 0.75 for one leg standing, 0.73 for functional reach, and 0.31 for tandem gait for the non-disabled sample. Reliability coefficients were 0.85 for one leg standing, 0.79 for functional reach, and 0.62 for tandem gait for the disabled sample. CONCLUSIONS: These findings for the one leg standing and functional reach testing protocols in disabled and non-disabled older people can be used as outcome measures. Further study should be directed towards improving the reliability of the tandem gait test for use with older people. PMID- 9859137 TI - The use of musculoskeletal techniques in adult cerebral palsy. PMID- 9859136 TI - Improvement in ejection fraction by hydrotherapy as rehabilitation in patients with chronic pulmonary emphysema. AB - BACKGROUND AND PURPOSE: It has been reported that breathing exercise by immersion in combination with expiring into water improved pulmonary function and blood-gas exchange in patients with chronic obstructive pulmonary disease (COPD). This effect may result from respiratory muscle exercise against hydraulic pressure and decreased dead space by increased sub-abdominal pressure. Increased cardiac function by bathing is suggested to contribute to the effect. To clarify the effect of the breathing exercise by immersion on cardiac function, a study was made of cardiac function during the exercise in patients with emphysema. METHOD: Echocardiography, respiratory function test and arterial blood gas analysis were performed during a two-month breathing exercise programme in a pool filled with 38 degrees C water for 12 patients (aged 70.9 +/- 9.1 years) with stable chronic pulmonary emphysema who were treated at our hospital between 1993 and 1996. The patients breathed in whilst standing in a pool and breathed out through the mouth while sinking the nose under water. This exercise was repeated for 30 min per day and continued for six days a week for two months. RESULTS: The ejection fraction increased significantly after each 30-minute exercise (N = 12; p < 0.01) as well as after the two-month exercise programme (N = 12, p < 0.05). Left ventricular end-diastolic and systolic dimensions at rest decreased significantly after the two-month exercise programme (N = 12, p < 0.01). The ratio of forced expired volume in one second to forced vital capacity (FEV1:FVC) increased and PaCO2 decreased following this programme (N = 12, p < 0.05). CONCLUSIONS: These results suggest that the breathing exercise by immersion is useful not only in treating emphysema but also in improving cardiac function. PMID- 9859138 TI - [Frontal displacement chromatography of melittin on meshed polymer ionites]. AB - Conditions of frontal displacement chromatography were optimized for preparative purification of the polypeptide hormone melittin from bee venom. Melittin was purified to an extent higher than that of a standard preparation described in the biochemical literature. PMID- 9859139 TI - [The use of polyelectrolytes for isolation of trypsin inhibitor from industrial waste of fractionation of alfalfa leaf proteins]. AB - Arypsin inhibitor was concentrated from the albumin fraction of alfalfa leaf juice as insoluble complex precipitate with pectin, carboxymethylcellulose sodium salt, and dextran sulfate. The concentration degree varied from 40 to 100, and the yield of the inhibitor was 45-50%. The homogeneous inhibitor was further isolated from solution of the precipitate by affinity chromatography on Trypsin Sepharose. The final product had a molecular weight of 6.9 kDa, high inhibitory activity (16.6 U per mg protein) and relatively high (about 30%) content of the alpha-helical form. PMID- 9859140 TI - [Production of human insulin from proinsulin by a biological catalysis]. AB - The conversion of porcine and human proinsulins to insulin by enzymatic hydrolysis induced by trypsin and carboxypeptidase B was studied. The conditions of reactions in the presence of trypsin (pure or immobilized in PAAG) and carboxypeptidase B were determined. The possibility of insulin production by the use of immobilized trypsin and carboxypeptidase B for practical purposes was shown. PMID- 9859141 TI - [Detection of DNA autoantibodies using a novel amperometric biosensor on the basis of platinum(II) complex with DNA]. AB - A biochemical biosensor was developed on the basis of a steady-state Hg-film electrode and either DNA molecules or antibodies to DNA immobilized in a cellulose nitrate film. This biosensor is designed to measure the concentration of DNA (or antibodies to DNA), and it can be used in the diagnosis of autoimmune diseases. Electric current of catalytic H2 evolution caused by complexing between DNA (antibodies to DNA) and Pt(II) was used as an analytical signal. The detection sensitivity threshold for DNA and antibodies to DNA was 10 and 0.075 microgram/ml, respectively. PMID- 9859142 TI - [The use of pancreatic RNAse in the production of baker's yeast]. AB - Microdoses of a preparation of pancreatic RNase were shown to stimulate the growth of Saccharomyces cerevisiae yeast. The effect was only retained at a certain inoculum/enzyme preparation ratio. Industrial tests of the preparation showed that the addition of RNase to the first reservoirs for culture accumulation (an inoculator and a seeding device) increased the yield of bakers' yeast and improved their quality. PMID- 9859143 TI - [Polyneuropathy in patients with periodic leg movements during sleep]. AB - INTRODUCTION: Periodic legs movements of sleep (PLMS) are rhythmic, standard and repetitive contractions of muscles of the extremities during the sleep. It is known that the patients with restless legs syndrome (RLS) have disorders during the sleep: increase in the latency of the sleep, increased number of arousal, etc.; most of them have also periodic movements of the legs during the sleep. OBJECTIVE: The relationship of the periodic movements of the legs during the sleep with polyneuropathy is not clear. Some authors have found evidence of electrophysiological and pathological of signs of axonal mild polyneuropathy in patients with restless legs syndrome. In this work, we evaluated nine patients that were diagnosed of PLMS, to determine the prevalence of neuropathy in such sample. METHOD: Polysomnography of nocturnal sleep of 7-8 hours was performed, including electromyographic recording of both anterior tibialis muscles; and electroneurographic study of peroneal, sural, ulnar and median nerves. DISCUSSION: Just in none of the nine studied cases were obtained electrophysiological signs of neuropathy; though it has been able to demonstrate the existence of mild alteration of the peripheral nervous system, fundamentally of sensory character; nevertheless, C we think that it would have to be studied the existence of polyneuropathy in all the patients with PLMS in order to discard potentially tractable organic causes. PMID- 9859144 TI - [Record of neurological emergencies at a tertiary care hospital]. AB - INTRODUCTION AND OBJECTIVE: There is a progressive increase in the demand for multidisciplinary attention from the Emergency Medical Services. The objective of this study was to determine the proportion of neurological conditions in the total hospital emergency workload, their demographic composition and medical needs. PATIENTS AND METHODS: We present an observational study of a cohort of histories of neurological emergencies at a tertiary hospital during a period of one year. RESULTS: On analysis of all emergencies, 1,592 were neurological conditions. The duty neurologist was consulted in 87.9% of the cases. The average age was 59, and the majority were women (56.3%). Maximum demand was between 14.00 and 19.59 hours. Monday was the day of the week when most were seen. The commonest causes of consultation were change in strength or language, headache and epileptic crises. The complementary investigation done most frequently was a blood test. Cerebral CT scan were done in 31.9% of the patients. The diagnoses most often made, as a group, were cerebrovascular disease followed by epilepsy and headache. The majority of the patients were referred to Primary Care Centres or for neurological consultation. When the patients were grouped according to the specialties with doctors on duty, the second largest group were seen by the emergency neurologist at our hospital. CONCLUSION: In view of the above results, we consider the presence of a neurologist in the Emergency Department to be fully justified. PMID- 9859145 TI - [Morbidity due to intracerebral hemorrhage]. AB - INTRODUCTION: Intracerebral hemorrhage (HIC) represents 10-30% of all stroke. Epidemiological studies have shown factors associated with its high mortality, but those which might lead to lower morbidity are little known. OBJECTIVE: To find the factors which may influence the functional state of a series of patients with intracerebral hemorrhage. PATIENTS AND METHODS: We made a prospective study of 203 patients with intracerebral hemorrhage. We recorded vascular risk factors, arterial blood pressure, laboratory and neuroimaging parameters obtained on admission. The functional condition when discharged from hospital was evaluated on the Rankin Scale, in three categories: independent (Rankin 0-1), partially dependent (Rankin 2-3) and totally dependent patients (Rankin 4-5). RESULTS: There was a 23.2% death rate. Of the 156 patients who survived, at the time of hospital discharge 35.8% had a score of 0-1, 50.6% a score of 2-3 and 13.4% a score of 4-5 on the Rankin Scale. Age (p < 0.005), hyperglycaemia (p < 0.05) and size of hemorrhage (p < 0.05) were associated with increased morbidity. CONCLUSIONS: The functional condition on hospital discharge was better in younger patients with lower levels of glycemia on admission and smaller hematomas on CT. PMID- 9859146 TI - [Swallowing changes in cerebrovascular accidents: incidence, natural history, and repercussions on the nutritional status, morbidity, and mortality]. AB - OBJECTIVE: To determine the frequency of dysphagia in CVA, its natural history and value as a risk factor of respiratory infection, malnutrition and death. PATIENTS AND METHODS: A prospective study was made of 187 consecutive patients with cerebrovascular accidents (CVA). A standardized test for dysphagia was done during the first two days of the illness and repeated three days a week. The levels of urea, total proteins and albumin were determined on admission and on discharge. The patients were questioned by phone after 6 months. RESULTS: There was dysphagia of liquids in 36.4% of the patients. The incidence of dysphagia for semisolids was of the same frequency but more severe. Coma was the cause of inability to swallow in 25.7% of the patients. During their stay in hospital one third of the patients with dysphagia died, one third became normal and one third still had dysphagia when they were discharged. After one week, one, three and six months respectively, the cure rate for dysphagia was 29.4%, 4.1%, 55.9% and 55.9%, and survival 83.8%, 67.6%, 61.8% and 60.3%. Thus after 6 months only 3 patients (4.4%) were alive and dysphagic. Half of the 'cures' occurred in the first week, and none occurred after more than 77 days. As compared to the non dysphagic patients, the dysphagic patients had 10 times more risk of respiratory infection, 18 times higher risk of death, greater loss of albumin and less loss of urea. CONCLUSIONS: There is a high prevalence of dysphagia in CVA and although functional prognosis is not unfavorable, respiratory infections, malnutrition and death are frequent. PMID- 9859147 TI - [New interactive verbal dichotic listening test]. AB - INTRODUCTION AND OBJECTIVE: From the multimedia technology and of the digital recording of the voice, we have redesigned the verbal dichotic listening test. The recognition of 15 monosyllabic word pairs offered though some earphones is presented in an interactive program. METHODS: Using the mouse button, the subject decides the moment in which each stimuli coupe will take place. Likewise, the screen displays 10 words, among which you have to recognize the words supplied by the earphones. Finally, the program results in complete right answers and right answers cross or referred to the opposite ear. RESULTS: Applied the test to a sample of 92 subjects, apparently normal, with a middle and high cultural level, from among 19 and 60 years (44 females and 48 males), the test resulted in a predominance of rights answers in the right ear (p < 0.0064). Separated the sample in in left-handed and right-handed men (67 right handed and 25 left handed), it hasn't been found meaningful differences among their averages. Separated the sample by sex, the differences among the slightly meaningful averages (p < 0.04, according to the Student's t test). CONCLUSIONS: The test is offered as a resource appropriate for laterality studies, and for differential disorder study of gnosias and praxias of the dyslexias of aphasias, hearing aids, multiple sclerosis, and aptitudes studies. PMID- 9859148 TI - [Oral anticoagulation in the secondary prevention of cerebrovascular disease. Long-term follow-up of 169 patients]. AB - INTRODUCTION: Although the indications for oral anticoagulation (AO) in the treatment of cerebral vascular disease (CVD) are well established, their potential side effects continue to give cause for worry. OBJECTIVES: To describe the complications and ischemic relapses in patients treated with AO for secondary prevention of CVD of cardiac embolic origin. PATIENTS AND METHODS: We included 169 patients with embologenic cardiopathy who, following an CVD, were treated with AO and followed-up at our medical centre for at least three months. We recorded their past clinical history and risk factors, occurrence of vascular relapses (VR), complications involving hemorrhage (CH), and data regarding course and follow-up. RESULTS: During an average follow-up of 50.3 months of a total of 707.9 patient/years, 20 VR (2.8% per year) were recorded; 15 of these were cerebro-vascular and mainly mild. We recorded 59 CH in 41 patients (8.3% per year) of which 6 were considered to be major. There was a 30% drop-out rate from follow-up at our centre, mainly due to death from other causes or to change of referral centre. CONCLUSIONS: There is a low incidence of relapse and of complications (usually mild) following AO for the secondary prevention of CVD of cardio-embolic origin. Efficacy and security are maintained in the long term. PMID- 9859150 TI - [Increase in the incidence of occipital plagiocephaly]. AB - INTRODUCTION: The diagnosis and treatment of posterior plagiocephaly is one of the most controversial aspects of craniofacial surgery. PATIENTS AND METHODS: The purpose of this study is to describe a recent increase in the incidence of occipital plagiocephaly without synostosis in our hospital during the last 6 months. The shift in the referral patterns is roughly contemporaneous with the American Academy of Pediatrics recommendations regarding infant sleep position. The temporal coincidence of this increase with the recommendation to avoid the prone sleeping position, to reduce the risk of sudden infant death syndrome, suggests a possible causal relationship. If the association is causal, education regarding the need of head position rotation coupled with that for sudden infant death syndrome should obviate positional occipital plagiocephaly. CONCLUSIONS: The feature of true lambdoid synostosis versus those of deformational plagiocephaly secondary to positional molding are inadequately described in the literature and poorly understood; the differential diagnosis is important in relation to a conservative diagnostical and therapeutical intervention in patients with positional molding. PMID- 9859149 TI - [Intracranial epidermoid and dermoid cysts]. AB - INTRODUCTION: Dermoid and epidermoid cysts are dysembryogenic tumors representing 1% of all intracranial tumors. They are characterized by slow growth, localization to the middle in relation to the basal cisterns and their benign course. OBJECTIVES: To analyze the clinical and radiological characteristics, and the results of surgical treatment of intracranial dermoid and epidermoid cysts. PATIENTS AND METHODS: We reviewed 18 cases of epidermoid and 3 dermoid intracranial cysts which had been diagnosed and treated during a period of 18 years. All patients were diagnosed by computerized tomography (CT) and the last 15 cases by magnetic resonance (MR). RESULTS: The commonest site was the temporal lobe with 7 cases, followed by the suprasellar region with 4 cases. All patients had peculiar imaging characteristics both on CT (hypodense and with no contrast silhouetted) and on MR (hypointensity in T1 and in T2). They were completely resected in 16 cases. There were 3 episodes of aseptic meningitis and 2 deaths. CONCLUSIONS: Dermoid and epidermoid cysts are characterized by long term clinical symptoms. The commonest site is supratentorial in the midline or in the temporosylvian region. The diagnostic investigation of choice is MR. Treatment is surgical, preferably complete resection. However, after sub-total resection there is a prolonged period of remission. PMID- 9859152 TI - [Neurophysiological parameters and CD4 lymphocyte count in patients infected by human immunodeficiency virus]. AB - INTRODUCTION AND OBJECTIVE: The human immunodeficiency virus (HIV) causes serious, irreversible, progressive deterioration of the immune and nervous systems. The main target cells are the 'helper' T lymphocytes and monocyte macrophage cells with CD4 molecules on the surface acting as virus receptors. In this study we attempt to find whether the immune state and the nervous system are involved in parallel, or whether, on the contrary, HIV neurotropism is such that it leads to early nerve involvement, out of proportion to that of the immune system. PATIENTS AND METHODS: We studied a total of 66 persons, 30 seronegative and 36 seropositive, with different CD4 lymphocyte counts. In all cases motor and sensory conduction studies were done in the arms and legs, namely auditory, visual and somatosensory evoked potentials and also endogenous potentials (mainly P300). CONCLUSIONS: There are neurophysiological parameters which give pathological figures, even when immunity is maintained (figures of CD4 greater than 500/mm3) especially with regard to the figures for sensitivity of the legs, somatosensory evoked potentials and P300. Moreover, these are increased and others added at the same time as the CD4 count falls as the disease advances. PMID- 9859151 TI - [Serum lipids. lipoproteins, and apolipoproteins in adult epileptics treated with carbamazepine, valproic acid, or phenytoin]. AB - INTRODUCTION: It is known that anticonvulsants are able to modify lipid profile; nevertheless the initial studies were performed in patients or polytherapy whereas later investigations were carried out mostly in children. OBJECTIVES: To evaluate serum lipids, lipoproteins and apolipoproteins in adult epileptic patients on monotherapy. MATERIAL AND METHODS: Total cholesterol, triglycerides, low- and high-density lipoprotein cholesterol (including the HDL2 and the HDL3 subfractions) and apolipoproteins A1 and B were measured in 120 epileptics patients treated with carbamazepine (n = 42), sodium valproate (n = 38) and phenytoin (n = 40) and compared with the values of 48 healthy subjects. RESULTS: Most of the measured parameters were significantly higher in patients receiving carbamazepine or phenytoin; carbamazepine-treated subjects showed specifically an increase in HDL2 lipoprotein cholesterol, whereas phenytoin-treated subjects showed specifically an increase of triglycerides; all of the observed alterations, save the increase in HDL lipoprotein cholesterol and apolipoprotein A1, were significant in women but not in men. Carbamazepine nor phenytoin related changes showed any correlation with the dose or the plasma levels of the drugs. No relevant modifications of serum lipids were seen in patients who received sodium valproate. CONCLUSIONS: The observed alterations in serum lipids were associated to the use of anticonvulsants with enzyme inducing activity and showed significant differences between both sexes. PMID- 9859153 TI - [Clinical study of neurofibromatosis type 1]. AB - INTRODUCTION: Recklinghausen's disease is considered to be the autosomal dominant disorder with the highest rate of mutation after achondroplasia. It is a neuroectodermal disorder with considerable clinical effects. PATIENTS AND METHODS: We present a study of 14 patients seen for cafe-au-lait spots in the Clinical Genetics Department of the Hospital Infantil Sur. A detailed questionnaire and physical examination was done to obtain a clinical outline. CONCLUSION: Suspicion of this condition, together with laboratory investigations led to the conclusion that the cases were neurofibromatosis. PMID- 9859154 TI - [Cerebral evoked potentials and semantic categorization]. AB - INTRODUCTION: Cerebral evoked potentials represent variations in the electrical activity of the nervous system, excited by a sensory stimulus and which are recorded on the surface. They may be classified as endogenous or exogenous. Amongst the late endogenous evoked potentials (EEP) we may emphasize N400 which seems to represent the linguistic management wave, particularly for semantics. OBJECTIVE: To observe the EEP recorded throughout the process of carrying out a task involving sequential semantic categorization. PATIENTS AND METHODS: Nine healthy subjects carried out a task based on a paradigm of sequential reaction time (RT), similar to that designed by us in 1994. The data recorded were the RT and EEP at the level of the electrodes Fz, Cz and Pz in five lots of stimuli (in the first four lots the same list of words were repeated and in the fifth and final lot a different, new list appeared). CONCLUSIONS: It was seen that only N400 varied during the procedure. This variation was related to familiarity with the task. That is to say that the amplitude of N400 was reduced when the same words were repeated and increased when new words appeared for semantic classification. The reduction in parallel, although not significant for RT, allowed the reduction in N400 alone to be followed whilst the task was carried out. It may be the electrophysiological marker of the process of learning semantic categorization. PMID- 9859155 TI - [Analysis of the changes observed in the sleep EEG recording of patients with juvenile myoclonic epilepsy]. AB - OBJECTIVE: The initial phases and transitional periods of sleep facilitate electroclinical manifestations of juvenile myoclonic epilepsy (JME). The period at the time of waking is identified in the different phases of sleep by cyclic electrophysiological oscillations, which in turn are synchronous with the spike and-wave or multiple spike-and-wave activity of the electric crises seen in JME and other epileptic syndromes. This observation supports the theory of an alternating cyclical pattern (ACP) as the trigger of these discharges. PATIENTS AND METHODS: Following classical macrostructural and microstructural patterns of EEG sleep analysis, we investigated electrophysiological behavior in 13 patients diagnosed as having JME, and 5 persons of homogeneous epidemiological characteristics as controls. RESULTS: In our study we observed that both groups had similar macrostructural sleep parameters. However, in patients prone to crises, these were concentrated in the first two hours recorded and showed no differences in sleep-waking transition or ACP of phase I sleep. These were significant when both periods were compared with the remaining NREM sleep. CONCLUSION: We conclude from this study that ACP is the microstructural element of sleep which modulates and permits classification of epileptiform anomalies, mainly in studies at the time of wakening. PMID- 9859156 TI - [Lipids and lipoproteins in a group of patients with ischemic cerebrovascular disease]. AB - OBJECTIVE: To study the blood levels of a group of lipids and lipoproteins in patients with ischemic cerebrovascular disease which was not secondary to cardiac embolic disease. PATIENTS AND METHODS: We assessed 40 patients of an average age of 64.5 years. Of these, 26 had cerebral infarcts due to disease of the great vessels and 14 had lacunar infarcts. Forty persons with no cerebrovascular disease were used as controls. We excluded patients diagnosed as having renal failure, liver, hematological, neoplastic and acute febrile disorders. Plasma was analyzed for total cholesterol, triglycerides, lipoproteins (a), HDL, LDL, VLDL and apoprotein B between 3 weeks and 6 months after the initial stroke. RESULTS AND CONCLUSIONS: Levels of triglycerides and of lipoprotein (a) were significantly greater in patients than in controls. We found no differences between the levels of total cholesterol, HDL, LDL, VLDL in the three groups studied. However, apoprotein B was greater in the controls than in the patients. There were no significant differences between the groups with lacunar ictus and those with infarcts secondary to disease of the great vessels. PMID- 9859157 TI - [Usefulness of multimodal evoked potentials and the electroretinogram in the early diagnosis of brain death]. AB - INTRODUCTION AND METHODS: We studied 72 patients fulfilling all the criteria for brain death (BD) by means of a series of tests: brain stem auditory visual evoked potentials (VEP), short latent period somatosensory evoked potentials (SEV) and electroretinogram (ERG). RESULTS: Three characteristic PEATC patterns were identified: bilaterally flat (73.34%), bilateral I wave (16.66%) and unilateral I wave (10%). The N20 wave of the SEV and other later cortical components were not seen in any patients, whilst the so-called subcortical components were partially or totally maintained. The use of non-cephalic data permitted discussion as to the origin of the subcortical components. When cephalic data was used to obtain the ERG and the VEP, the a and b waves of the ERG were recorded in all cases, whilst in the VEP channel waves of inverse polarity appeared of similar morphology and the same latency, but of lower amplitude than those of ERG. When a non-cephalic reference was used, the morphology and latency of the ERG components were unchanged, whilst no response was obtained from the VEP channel. This electrophysiological pattern indicates that the only part of the visual pathway of patients with BD to maintain electrical activity is the retina. CONCLUSION: The application of this set of tests permits early diagnosis of BD, which is basic for transplantation. PMID- 9859158 TI - [Somatosensory trigeminal evoked potentials after gingival stimulation of the mental nerve. Normal values]. AB - OBJECTIVE: Somatosensory evoked potentials were recorded following unilateral stimulation of the mental nerve in the gum of 100 healthy volunteers aged between 17 and 22 years. METHODS AND RESULTS: Responses were recorded until 100 ms with electrodes placed over the scalp (C5/C6) referenced to central frontal (Fz). In 10 subjects, simultaneous recordings were made in masticatory and facial muscles to detect possible muscle artefacts. Stimulation was effected using a specially designed stimulator adaptable to each individual. Contralateral responses consisted of four very constant deflexions (N12, P19, N26 and P35) forming a W shaped complex of mean duration 31.27 ms. Tables of normality were compiled for latencies and amplitudes with confidence intervals of 99.8% reliability. Constancy of deflexions, stability of response (by serial studies), and possible sexual differences were also studied. Muscle artefacts were ruled out, and the participation of the mental nerve in the genesis of the responses was confirmed. CONCLUSION: We consider the proposed method a reliable alternative to other procedures used to obtain TEPs. PMID- 9859159 TI - [Amyotrophic neuralgia: review of 37 cases]. AB - INTRODUCTION: Amyotrophic neuralgia is characterized by pain of acute or subacute onset, accompanied by weakness and occasionally by atrophy of the brachial muscles, of unknown origin. We present our experience over the past 20 years. PATIENTS AND METHODS: We made a retrospective review of 37 patients with the above diagnosis, following the criteria of other series of such cases published in the literature. RESULTS: Twenty four of the patients were men and thirteen were women. The average age was 38 (11 to 71). A relevant clinical history was recorded in 9 cases; infection (5), surgery (4), remote trauma (3) and vaccination (1). There was a painful onset of the condition in 32 patients; objective weakness of the superior brachial plexus (30), inferior (5) or both (2). Atrophy was present in 23 and hypoaesthesia in 13. Two patients had fasciculations and 9 had hyperreflexia. In all patients electromyographic studies showed a neurogenic pattern of denervation of the muscles clinically affected. The severity of the condition was divided into mild (18), moderate (16) and intense (3). Prognosis was good in 24 and sequelae remained in 11. There were 2 bilateral cases and 2 relapses but no familial cases. CONCLUSIONS: There was a ratio of men/women of 1.8:1 and onset usually when the patient was in his forties. Mild infection, surgery, remote trauma and vaccination were the commonest clinical factors. Onset was painful in 85%. Muscular weakness was predominantly in the superior brachial plexus (85%), followed by atrophy in 62%. There was hypoaesthesia in a third of the patients. Most cases were mild (50%) and made a complete recovery (70%). Our findings are similar to those described in most series in the literature. PMID- 9859160 TI - [Statins (HMG-CoA reductase inhibitors), cholesterol and stroke]. AB - The HMG-CoA reductase inhibitors (known as statins) have been shown to reduce morbi-mortality of vascular origin, including transitory ischemic accidents and cerebral infarcts, in a large group of patients. The implications, both clinical and of costs, oblige us to consider analysis of the existing evidence and the answers to a number of questions arising in a field which has not yet received much attention from neurologists. PMID- 9859161 TI - [Intensive neurology. Past, present, and future]. AB - Although the neurological intensive care, seem have originated at the ends of 40s, during the epidemic of acute poliomyelitis that flogged Europe, it must be indicated that the growth and expansion of this subspecialty, has been a remarkable fact only in the course of the two last decades. Despite the fact that the neurological intensive care units (Neuro-ICU) are expensive; multiple have been the benefits derived from their creation; so much for patients, hospitals, as well for medical teaching. This is the current panorama of these units, mainly in developed countries, however, unfortunately this is not the situation in others, especially the underdeveloped ones. Many of the dilemmas that today confronts neurology in our countries are due to the nonexistence of these units. Undoubtedly, the neurocritical patients results more benefitted, when receives attention from the neurointensivists; thus the medical care that it receives becomes defragmented. The creation of the Neuro-ICU in our countries should not be made in a generalized way, but strategically, in addition, would be very convenient the incorporation of neurointensivists in the polyvalent intensive care units or intermediate care units. For the future, it will have to keep in mind the fact, that certain novel procedures that today emerge for the management of certain neurocritical conditions, will have to be assimilated by neurointensivists, since they will be the personnel disposed to implement them in any moment, and what is more important, it is the competent personnel prepared to treat any complication that emerge upon applying these. PMID- 9859162 TI - [Wegener's granulomatosis with meningeal involvement: clinic-radiological correlations]. AB - INTRODUCTION: Wegener's granulomatosis is a systemic vasculitis which, in its classical form, is characterized by involvement of the superior and inferior respiratory tract and the kidneys. The vasculitis may be multisystemic. Ophthalmic and neurological involvement are common (22% and 54% of those affected respectively). When considering involvement of the nervous system, the commonest finding is peripheral neuropathy, particularly in the form of multiple mononeuritis. Meningeal involvement is exceptional. CLINICAL CASE AND CONCLUSIONS: We present a case of Wegener's granulomatosis with meningeal involvement, studied using CT and MR. The findings using imaging techniques are described, and conditions which should be considered in the differential diagnosis are discussed. PMID- 9859163 TI - [Brown-Sequard syndrome as the initial clinical manifestation ofmultiple sclerosis]. AB - Multiple sclerosis is a disorder with different forms of clinical presentation. The Brown-Sequard syndrome has occasionally been described in association with multiple sclerosis. We present the case of a patient whose initial clinical presentation of multiple sclerosis was the Brown-Sequard syndrome. PMID- 9859164 TI - [Cleidocranial dysostosis. Presentation of a case]. AB - INTRODUCTION: Cleidocranial dysostosis is a syndrome defined by three characteristic findings: clavicular aplasia, retarded cranial ossification, and autosomic dominant hereditary transmission, with completed penetrance and full expression. However, the diagnosis cannot only be made based on those finding, because the polymorphism and extension of the lesions of this disease is important. Therefore, in this disease we can see upset in the second teething, short stature or dwarf, persistence of the biconvex appearance of vertebral body, bone hypoplastic iliac, retarded pubis branch ossification, wedge shape distal phalanges or with brachymesophalangia of the forefinger and fifth finger. CLINICAL CASE: We describe a 20 years old man, with cleidocranial dysostosis, without familiar antecedent (probable mutation), that come to our center for treatment of denture pathology with disabled eating, because anomalous distribution and eruption. He had clavicle agenesis, cranial ossification upset with wormian bones, vertebral bodies biconvex, superior maxillary hypoplastic, and dental packed in the superior maxillary and jawbone. CONCLUSIONS: Cleidocranial dysostosis is a hereditary disease, which can be of spontaneous apparition (mutation), has a grand polymorphism, affect the osseous development, predominate in the middle line membranous bone and is an entity of radiologic diagnosis. PMID- 9859165 TI - [Sleep-arousal dissociation in a case of hypersomnia]. AB - INTRODUCTION: Dissociated sleep-arousal states are clinical and experimental phenomena which represent mixed forms of the three stages of this cycle (arousal, NREM sleep and REM sleep). CLINICAL CASE: We describe the case of a man who presented with a history of excessive diurnal somnolence for the previous 5 years. He also had symptoms of sleep paralysis, hypnagogic hallucinations, automatic behavior and excessive movements during sleep. He had had no episodes of loss of muscle tone; MR was normal; HL DR2 and DQwl antigens were negative; two polisomnographic and a Multiple Latency Sleep test showed: 1. Absence of respiratory disorders. 2. Normal latency of REM sleep. 3. Periods of dissociated sleep (REM without atonia and arousal with atonia), and 4. An average sleep latency of 3.2 minutes and absence of REM periods. CONCLUSION: This case adds a new type of dissociated sleep state which may accompany the disorder known as hypersomnia without REM periods. PMID- 9859166 TI - [The nature, diagnosis and treatment of post-concussion syndrome]. AB - INTRODUCTION: The relationship between brief loss of consciousness, subsequent cognitive and emotional complaints, and impact on daily functioning continues to be hotly debated. DEVELOPMENT: In this paper the strong variability about prevalence of the postconcussional syndrome found in several studies is outstanding and the main issues of this disagreement are suggested. Recent neuroimaging techniques are discussed and some neuropsychological measures are suggested. CONCLUSIONS: Currents models (organic/psychogenic) of postconcussional symptoms are reviewed, and a multifactorial model which integrates biological factors with the relevance of neuropsychological deficits--attention, memory, speed of information processing--and coping process is proposed. Finally, according with this model, we conclude with some suggestions to improve neuropsychological intervention and medical treatment of these patients. PMID- 9859167 TI - [Neurological complications of aortic artery surgery]. AB - INTRODUCTION: Improvement in cardiac surgery techniques has led to a considerable reduction in mortality following surgery of the aorta, dissection of the aorta and for aortic aneurysms. Although there are satisfactory vascular results following surgical repair, morbi-mortality related to neurological complications, both cerebral and spinal, is still very high. Repair of aortic lesions may lead to damage by two main mechanisms: ischemia secondary to prolonged obstruction of the aorta or to neurological lesions due to total circulatory arrest. DEVELOPMENT: After description of the mechanisms leading to cerebral and spinal lesions and the risk factors involved, we discuss methods of vascular protection and other means of neuro-protection, both spinal and cerebral. Of the vascular techniques for spinal protection, the most useful ones are short-circuits, or active or passive shunts, and cardiopulmonary by-pass. Amongst the vascular techniques for cerebral protection we describe total circulatory arrest and methods of anterograde and retrograde cerebral perfusion, and when these may be used. As general measures for nervous system protection, we describe the use of hypothermia and drugs such as corticosteroids, free radical blockers, antagonists of the excitatory amino-acids etc. CONCLUSIONS: In spite of the number of studies done, the extreme sensitivity of the nervous system to ischemia has meant that in few cases have encouraging results been seen. Neurological damage continues to be the main cause of morbi-mortality in patients with dissection or aneurysm of the aorta. PMID- 9859168 TI - [Concept and historical evolution of epilepsy in pre-Colombian and Viceregency Peru]. AB - INTRODUCTION: We review and discuss beliefs regarding epilepsy, known in the Quechua language as sonko-nanay (disease of the heart). An overall view given by the indigenous people allows an outline to be drawn of tradition in Peru. DEVELOPMENT: This includes Quechua mythological beliefs, with understandable names and/or analogy with the different types of crises according to modern diagnostic classification, methods of treatment under the three different systems of government as well as real and fictitious surgery by trepanation of the skull. CONCLUSION: During the period of the Viceroys (based on Spanish medical knowledge) the different meanings are considered together with the confusion with tetanus and hysteria, and the outlines of treatment used. PMID- 9859169 TI - [Drug-resistant epilepsy]. PMID- 9859170 TI - [Painful ophthalmoplegia of sudden appearance in an elderly lady]. PMID- 9859171 TI - [Polyneuropathy in the critical patient. Relevance of neurophysiological studies]. PMID- 9859172 TI - [Intracranial arterio-venous malformations: an unresolved problem]. PMID- 9859173 TI - [Polite expressions in requests for neurophysiological studies]. PMID- 9859174 TI - [Familial and genetic cluster headache]. PMID- 9859175 TI - ["Trombolisis" or "trombolisis"]. PMID- 9859176 TI - Historical redistribution under the Social Security Disability Insurance Program. AB - This study uses Social Security administrative data on historical taxes and benefits by year, age, gender, and race for an ex post analysis of redistribution under the Disability Insurance (DI) program. The relationship between the taxes paid and benefits received to date under the program is described for successive cohorts as a whole and for specific race and gender groups both within cohorts and across time. PMID- 9859177 TI - Pension integration and Social Security reform. AB - Many employer-provided pensions plans explicitly account for Social Security in their benefit formulas-a practice known as integration. Because integrated pensions are directly linked to Social Security, both the incidence and design of explicitly integrated plans are likely to be affected by changes in the current Social Security program. While integration has been mentioned as an important issue in discussions of Social Security reform, researchers have largely ignored the concept of pension integration. This article provides basic information about pension integration and addresses, in general terms, the relationship between Social Security reform and pension integration. PMID- 9859178 TI - [The role of the striatum in the evolution of the mammalian forebrain]. PMID- 9859179 TI - [Study of the projections of the thalamic posterolateral nucleus to the field 18A of the rat extrastriatal cortex]. PMID- 9859180 TI - [NMDA-Induced oscillations of membrane potential in isolated spinal neurons from lamprey, Lampetra fluviatilis]. PMID- 9859181 TI - [GABA-Activated cloride currents in isolated spinal neurons from lamprey, Lampetra fluviatilis]. PMID- 9859182 TI - [Neuropeptide Y (NPY)-immunoreactivity in limbic and non-limbic structures of the telencephalon in turtles. Comparison with the literature data obtained in mammals]. PMID- 9859183 TI - [Structural and functional characteristics of supraspinal fibers monosynaptically connected with the lumbar motoneurons of the spinal cord from the frog, Rana ridibunda]. PMID- 9859184 TI - [Functional characteristics of correlation of the vestibular input and the vestibulospinal system of the frog, Rana ridibunda]. PMID- 9859185 TI - [Morphometric study of the development of the cerebellar Purkinje cells during postnatal rat ontogenesis]. PMID- 9859186 TI - [Effect of hypoxia on cell membranes of the right and left hemispheres of the rat embryo brain]. PMID- 9859187 TI - [Examples of recapitulation of the phylogenetic stages of development of the wakefulness-sleep cycle in mammalian ontogenesis]. PMID- 9859188 TI - [Disturbances in the wakefulness-sleep cycle and its dissolution]. PMID- 9859189 TI - [Effects of 6-hour deprivation of protosleep (primary sleep) in the brown frog, Rana temporaria]. PMID- 9859190 TI - [Study of intragroup competitive relations in the adolescent chimpanzee]. PMID- 9859191 TI - [Effect of 6-hour sleep deprivation in the guinea pig Cavia porcellus]. PMID- 9859192 TI - The teaching of crystallography: a historical survey. AB - The views of several well known crystallographers on how to teach the subject are presented with a description of teaching texts and schools available on a world wide scale. PMID- 9859193 TI - Electron crystallography--accomplishments and challenges. AB - Electron crystallography is a powerful tool for the quantitative structural characterization of substances that preferentially form thin microcrystals. Because multiple-beam dynamical scattering may cause observed diffraction intensities to deviate significantly from their kinematical values, it is necessary to demonstrate that the conditions favoring ab initio determinations can be established. Review of similar determinations made from electron and X-ray data make clear both the strengths and weaknesses of electron crystallography. With current instrumentation, the major onus now placed on the experimentalist is to optimize specimen preparation so that the resultant diffraction data can be directly interpreted. PMID- 9859194 TI - A molecular model for muscle contraction. AB - The molecular mechanism of muscle contraction has been elucidated by a combination of electron microscopy, biochemistry and X-ray diffraction from fibres and crystals. Protein crystallography provided the essential molecular anatomy for understanding this problem. Synchrotron radiation has played a crucial role. PMID- 9859195 TI - Time-resolved crystallography. AB - Time-resolved crystallography has been successfully applied on the time scale from seconds via milliseconds and nanoseconds to picoseconds on a variety of systems. This brief review largely deals with macromolecular systems, on which there has been substantial recent progress. The strategies for design of a successful experiment that eliminates or minimizes potential artefacts have been identified, and the specifically crystallographic components of these strategies have been implemented. The remaining computational challenge is to identify and extract time-independent structures, each corresponding to a distinct reaction intermediate, whose populations vary with time and give rise to the time dependent X-ray diffraction data. The fourth dimension, time, has been added to the three spatial dimensions of crystallography; it can no longer be regarded as purely a static technique. PMID- 9859196 TI - Crystal growth and crystallography. AB - Selected topics that may be of interest for both crystal-structure and crystal growth communities are overviewed. The growth of protein crystals, along with that of some other compounds, is one of the topics, and recent insights into related phenomena are considered as examples of applications of general principles. The relationship between crystal growth shape and structure is reviewed and an attempt to introduce semiquantitative characterization of binding for proteins is made. The concept of kinks for complex structures is briefly discussed. Even at sufficiently low supersaturations, the fluctuation of steps may not be sufficient to implement the Gibbs-Thomson law if the kink density is low enough. Subsurface ordering of liquids and growth of rough interfaces from melts is discussed. Crystals growing in microgravity from solution should be more perfect if they preferentially trap stress-inducing impurities, thus creating an impurity-depleted zone around themselves. Evidently, such a zone is developed only around the crystals growing in the absence of convection. Under terrestrial conditions, the self-purified depleted zone is destroyed by convection, the crystal traps more impurity and grows stressed. The stress relief causes mosaicity. In systems containing stress-inducing but poorly trapped impurities, the crystals grown in the absence of convection should be worse than those of their terrestrial counterparts. PMID- 9859197 TI - Crystal structural studies of a membrane protein complex, cytochrome c oxidase from bovine heart. AB - Membrane proteins invest the membranes of living cells and organelles with such functions as energy conversion, molecular transport and signal transduction. The number of known structures of membrane proteins is less than 20, which is too few to understand the highly organized biological reactions in a cell. The whole procedure, including crystallization, for bovine heart cytochrome c oxidase is described as an example of crystal structural analysis of a membrane protein. Structural features of membrane proteins and structure-function relationships of the enzyme are also discussed. PMID- 9859198 TI - Crystallographic studies on the ribosome, a large macromolecular assembly exhibiting severe nonisomorphism, extreme beam sensitivity and no internal symmetry. AB - Crystals, diffracting best to around 3 A, have been grown from intact large and small ribosomal subunits. The bright synchrotron radiation necessary for the collection of the higher-resolution X-ray diffraction data introduces significant decay even at cryo temperatures. Nevertheless, owing to the reasonable isomorphism of the recently improved crystals of the small ribosomal subunits, reliable phases have been extracted at medium resolution (5-6 A) and an interpretable five-derivative MIR map has been constructed. For the crystals of the large subunits, however, the situation is more complicated because at higher resolution (2.7-7 A) they suffer from substantial radiation sensitivity, a low level of isomorphism, instability of the longest unit-cell axis and nonisotropic mosaicity. The 8 A MIR map, constructed to gain insight into this unusual system, may provide feasible reasoning for the odd combination of the properties of these crystals as well as hints for future improvement. Parallel efforts, in which electron-microscopy-reconstructed images are being exploited for molecular replacement studies, are also discussed. PMID- 9859199 TI - Local genetic structure in red grouse (Lagopus lagopus scoticus): evidence from microsatellite DNA markers. AB - Allelic variation at seven hypervariable tri- and tetranucleotide microsatellite loci was used to determine levels of population differentiation between 14 populations of red grouse (Lagopus lagopus scoticus) in northeast Scotland, UK. Despite the potential for long-distance dispersal in grouse, and a semicontinuous habitat, significant population divergence was observed (mean RST = 0.153; P < 0.01) and an isolation-by-distance effect detected (Mantel test: P < 0.001). Examination of the spatial trend in principal component scores derived from allele frequencies among populations highlighted a barrier to gene flow that was confounding a simple isolation-by-distance effect. This barrier corresponded to an area of unsuitable habitat for grouse associated with a river system that bisected the study area. Mean genetic relatedness was higher for males than for females in all but one of the study populations, suggesting that the territorial behaviour and natal philopatry displayed by cocks have a manifold effect in generating the observed spatial genetic structure. Lower female relatedness values suggest a higher level of female-mediated gene flow, which is sufficient to prevent the loss of genetic variation from within populations and the onset of inbreeding effects. The potential consequences of local subdivision for red grouse populations are discussed. PMID- 9859203 TI - Population genetic structure, phylogeography and spawning philopatry in walleye (Stizostedion vitreum) from mitochondrial DNA control region sequences. AB - Mitochondrial (mt) DNA control region sequences were used to test the genetic and phylogeographic structure of walleye Stizostedion vitreum populations at different geographical scales: among spawning sites, lake basins, lakes, and putative glacial refugia in the Great Lakes region. Sequencing 199 walleye revealed nucleotide substitutions and tandemly repeated sequences that varied in copy number, as well as in sequence composition, in approximately 1200 bp of the mtDNA control region. Variable numbers of copies of an 11-bp tandem repeat showed no geographical patterning and were not used in further analyses. Substitutions in the other areas of the control region yielded 19 haplotypes, revealing phylogeographic structure and significant differences among glacial refugia, lakes, basins and some spawning sites. Differences among spawning populations were consistent with reduced gene flow, philopatry and possible natal homing. Analysis of spawning populations showed consistency of genotypic frequencies among years and between males and females, supporting philopatry in both sexes. The unglaciated plateau in southern Ohio, USA housed a very different haplotype that diverged prior to the Missouri, Mississippi and Atlantic glacial refugia types. Haplotypes from the three refugia colonized the Great Lakes after retreat of the Wisconsin glaciers, and their present distribution reflects the geography of their prior isolation and differential colonization. Populations that became associated with spawning localities appear to have diverged further due to philopatry, resulting in fine-scale phylogeographic structuring. PMID- 9859204 TI - Polymorphic microsatellite DNA markers in red drum (Sciaenops ocellatus). PMID- 9859205 TI - Microsatellites in reindeer, Rangifer tarandus, and their use in other cervids. PMID- 9859206 TI - Isolation, variability, and cross-species amplification of polymorphic microsatellite loci in the family Mustelidae. PMID- 9859207 TI - Isolation and characterization of microsatellite markers in the nuclear genome of the brown alga Laminaria digitata (Phaeophyceae). PMID- 9859208 TI - Microsatellite markers for the phalangerid marsupial, the common brushtail possum (Trichosurus vulpecula). PMID- 9859209 TI - Microsatellite primers from Geospiza fortis and cross-species amplification in Darwin's finches. PMID- 9859210 TI - Isolation and characterization of microsatellite markers in the bloodsucking bug Rhodnius pallescens (Heteroptera, Reduviidae). PMID- 9859211 TI - Identification and characterization of nuclear, cleaved amplified polymorphic sequence (CAPS) loci in Irvingia gabonensis and I. wombolu, indigenous fruit trees of west and central Africa. PMID- 9859212 TI - Loss of LKLF function results in embryonic lethality in mice. AB - Lung Kruppel-like factor (LKLF) is a member of the Kruppel-like family of zinc finger transcription factors and is closely related to erythroid kruppel-like factor (EKLF), which is necessary for beta-globin gene expression. While EKLF is expressed exclusively in erythroid cells, LKLF is expressed temporally during early embryonic development and predominantly in the adult mouse lung. To understand the role this novel transcription factor plays in development as well as tissue differentiation and function, animals lacking LKLF were produced using gene targeting technology. Mice lacking LKLF die in utero between day 11.5 and 13.5 of embryonic life and exhibit retarded growth, craniofacial abnormalities, abdominal bleeding and signs of anaemia. Although the yolk sac erythropoiesis is normal in mutant embryos, in vitro fetal liver cultures of these embryos fail to give rise to erythroid cells. Expression of other erythroid specific genes such as EKLF, GATA1 and GATA3 is unaltered in these animals. These findings demonstrate the LKLF function is indispensable during normal embryonic development, and although both LKLF and EKLF recognize common DNA motifs, they do not substitute for each other. PMID- 9859213 TI - Improved transfection efficiency of chicken gonadal primordial germ cells for the production of transgenic poultry. AB - Electroporation is a common method of DNA transfection for many types of eukaryotic cells, but has not been attempted in avian primordial germ cells (PGCs). DNA uptake in chicken primordial germ cells (PGCs) was tested using electroporation with and without dimethyl sulfoxide (DMSO). Gonadal tissue and chicken embryonic fibroblasts (CEFs) were isolated from 6-day-old embryos (stage 29), transfected with pCMV beta carrying the bacterial lacZ gene, and cultured for 24 h. Gonadal primordial germ cells (gPGCs) were purified from culture using a Ficoll gradient. The addition of DMSO significantly increased the transfection efficiency of gPGCs but had no effect on chicken embryonic fibroblasts. Electroporation of gPGCs resulted in an 80% transfection efficiency compared with about 17% observed with liposomes. Approximately 200 transfected gPGCs were injected into 2.5-day-old (stage 17) recipient embryos and the eggs were incubated for an additional 3.5 days, 7.5 days or to hatching. The exogenous gene was detectable in 100%, 67% and 41% of the 6-day-old (stage 29), 10-day-old (stage 36) recipient embryos and hatched chicks gonads, respectively. PCR analysis of DNA from the hatched chicks showed that exogenous lacZ DNA was detected only in the gonad and not the liver and heart. These results indicated that electroporation was a suitable means of transfecting avian gPCGs for the goal of producing transgenic poultry. PMID- 9859215 TI - Coat protein-mediated resistance to pea enation mosaic virus in transgenic Pisum sativum L. AB - Pea (Pisum sativum L.) plants were transformed in planta by injection/electroporation of axillary meristems with a chimeric pea enation mosaic virus (PEMV) coat protein gene contruct. R1 progenies of these plants were shown to harbor the transgene by polymerase chain reaction (PCR) and genomic Southern analysis, while transgene expression was demonstrated by western blot analysis. Transgenic R2, R3 and R4 plants displayed delayed or transient PEMV multiplication and attenuated symptoms as compared to control inoculated individuals. PMID- 9859214 TI - Lactation-induced WAP-SV40 Tag transgene expression in C57BL/6J mice leads to mammary carcinoma. AB - Two transgenic lineages were generated by directing the expression of SV40 T antigen to the mammary gland of inbred C57BL/6J mice using the whey acidic protein (WAP) promoter. In one lineage, WAPTag 1, multiparous female mice developed mammary adenocarcinoma with an average latency period of 13 months. The histopathological phenotype was heterogeneous, tumours occurred in a stochastic fashion, normal tissue was located next to neoplastic tissue, the mammary tumours usually developed and were remarkably similar to that observed in human cases. In addition, male and virgin females developed a poorly differentiated SV40 T antigen-positive soft tissue sarcoma, also at 13 months of age. In the other lineage, WAPTag 3, some parous females developed mammary tumours, but most mice succumbed to osteosarcomas arising from the os petrosum at 5.5 to 6 months of age and on necropsy, renal adenocarcinomas were also found. Appearance of these unexpected tumour types demonstrates the non-specific expression of SV40 Tag under the control of the WAP promoter. The expression of SV40 Tag in mammary glands at different stages of development was also examined, and only actively lactating glands were positive. This suggests that the abundant cyclic synthesis of SV40 Tag associated with pregnancy is required for mammary tumorigenesis in these lineages. PMID- 9859216 TI - Expression of a wool intermediate filament keratin transgene in sheep fibre alters structure. AB - Alteration of the protein composition of the wool fibre via transgenesis with sheep wool keratin and keratin associated protein (KAP) genes may lead to production of fibre types with improved processing and wearing qualities. Using this approach, we have demonstrated that high level cortical-specific expression of a wool type II intermediate filament (IF) keratin gene, K2.10, leads to marked alterations in both the microstructure and macrostructure of the wool fibres, which have higher lustre and reduced crimp. Analysis of mRNA found reduced levels of transcripts from endogenous cortical type I (p < 0.05) and type II (p < 0.01) keratin IF genes and from the KAP8 (p < 0.001) and KAP2 (p < 0.01) gene families. Examination of protein composition revealed an altered ratio in the keratin type II protein family of the wool fibre cortex. Whilst the over-expressed K2.10 transgene product constituted the majority of keratin type II IF protein, it appeared unable to form heterodimers with much of the expressed endogenous keratin type I IF. In comparison with non-transgenic sheep, fewer IF microfibrils were visible in the cortical cells of fibres from transgenics. The combined effect on fibre structure was disruption of the formation of orthocortical and paracortical cells in the fibre cortex, a factor which could account for the reduction in fibre crimp. No effects upon transcript or protein levels, or fibre microstructure or macrostructure were observed in transgenic sheep expressing the transgene at lower levels, indicating that subtle changes to the gene expression profile in sheep wool follicles can be tolerated. The data here also illustrate that control over endogenous transcript levels in the cortex results when factors acting on the endogenous keratin type I, keratin type II and KAP gene sequences are sequestered by the active K2.10 transgene locus. Moreover, interference to a transcriptional hierarchy shared by keratin and KAP genes may occur prior to establishment of the orthocortical and paracortical compartments of the follicle cortex, at the level of the chromatin. PMID- 9859217 TI - Overexpression of growth hormone affects alternatively spliced IGF-I mRNA expression in oMt1a-oGH transgenic mice. AB - Restorative growth hormone (GH) treatment of hypophysectomized rats differentially enhances the transcription of alternative IGF-I mRNA classes in liver. The goal of the present study was to determine the effects of GH overexpression on various classes of hepatic IGF-I mRNA in GH transgenic mice. Unstimulated oMt1a-oGH transgenic mice had low levels of transgene expression, and therefore were used to determine the effects of long-term, slightly elevated GH levels on the abundance on each alternative IGF-I mRNA class. The acute effects of high GH levels on the expression of alternative IGF-I mRNA were studied by gavaging transgenic mice with 25 mM zinc sulfate to activate oMt1a-oGH transgene expression. Long-term, low levels of oGH transgene expression in unstimulated transgenic mice resulted in a 73% down regulation of IGF-I 2Ea mRNA but not 1Ea and 2Eb mRNA. Acute stimulation of transgene expression triggered a rapid, 240% increase in 1Ea mRNA levels within 4 hours of transgene expression while 2Ea mRNA was down regulated to nearly non-detectable levels by 6 hours. IGF I 2Eb mRNA was not affected by the short-term GH elevation. Our results showed that IGF-I 1Ea and 2Ea mRNA were differentially regulated by chronic low or acute high levels of GH. These results suggest that the regulation of IGF-I 1Ea and 2Ea mRNA transcription involve different postreceptor molecules and/or feedback mechanisms. PMID- 9859218 TI - Nuclear localization signals enhance germline transmission of a transgene in zebrafish. AB - We report that cytoplasmic injection into zebrafish eggs of 10(4) copies of plasmid DNA complexed to nuclear localization signal (NLS) peptides, as compared to 10(6) copies of naked DNA, increased nuclear uptake of transgene DNA early during embryo development and enhanced transgene integration frequency into the germline of founders. Monitoring the dynamics of nuclear uptake of DNA-NLS complexes by fluorescence in situ hybridization (FISH) of interphase nuclei indicates that NLS enhances both the proportion of nuclei importing DNA during early embryo development, and the amount of DNA imported by individual nuclei. The use of NLS increases the proportion of germline transgenic founders from 14 to 43% (P < 0.01) as assessed by polymerase chain reaction analysis of F1s. From germline transgenic DNA-NLS-injected founders, 47% transgenic F1s are obtained in wild-type crosses, as opposed to 6% from naked DNA-injected founders (P < 0.01). In both cases, the transgene is transmitted to the F2 generation. In addition, high-resolution FISH analysis of transgenic F1s reveals that the use of NLS increases the number of distinct transgene integration sites along chromatin fibres. PMID- 9859219 TI - Expression of human erythropoietin transgenes and of the endogenous WAP gene in the mammary gland of transgenic rabbits during gestation and lactation. AB - An understanding of the expression of transgenes in the mammary gland during gestation and lactation is crucial for the use of transgenic mammals as bioreactors. Here we describe the temporal pattern of expression of the endogenous rabbit WAP gene and human erythropoietin (hEPO) transgenes under the control of rabbit WAP promoter and 3' flanking sequences. The endogenous rabbit WAP gene was expressed throughout gestation including the day of mating, as well as during lactation in transgenic rabbits bearing a minigene construct. In non pregnant cycling females, WAP expression was found independent of transgenic status; however, WAP expression was not detected in non-cycling females. The significance of this new finding is not clear at present. hEPO mRNA was detected in mammary gland biopsies from pregnant transgenic rabbits only on day 28 of gestation. During lactation, transcripts were present in mammary gland biopsy samples taken on days 0, 7, 14 and 21. A sharp decline in the levels of transcripts was found for an hEPOcDNA gene construct at the end of lactation (day 28). Although the levels of hEPO were too low to allow a conclusion to be drawn regarding temporal or position-dependent expression, this finding may reflect an integration position effect. PMID- 9859220 TI - A rapid and reliable PCR-based assay for gene transmission and sex determination in newborn transgenic mice. AB - This article describes a reliable and rapid method for simultaneous detection of a transgene and sex determination in the newborn mouse pups by PCR using three sets of primers in a single reaction. One set of sense/antisense primers is used to amplify the experimental transgene (androgen receptor gene in this case), the second set for the mouse Y-chromosome-specific SRY gene, and the third set for the beta subunit of the thyroid stimulating hormone (TSH beta), an internal control. This procedure allowed us to promptly analyze pups born from transgenic founders carrying the androgen receptor transgene and, at the same time, establish the sex of the animals. The method is simple, rapid and highly reproducible. PMID- 9859222 TI - Prediction of transgene integration by noninvasive bioluminescent screening of microinjected bovine embryos. AB - Transgenesis in domestic species, as a research tool and in biotechnological applications, has been limited by the expense of producing transgenic offspring by standard microinjection techniques. A major factor is the inefficiency of maintaining large numbers of recipient females, when a high percentage of these carry nontransgenic fetuses. There are two approaches to reduce this cost, the fusion of transfected fetal fibroblasts with enucleated oocytes, and the screening of microinjected embryos for transgene integration in blastocysts, prior to transfer. Here, we develop a luminescent screening system to select transgenic bovine embryos. A transgene with scaffold attachment regions flanking the murine HSP70.1 promoter linked to firefly luciferase cDNA, was microinjected into pronuclei of in vitro produced zygotes. At the blastocyst stage, the transgene was induced by heat shock (45 degrees C, 15 min) and 4-6 h later, luciferase expression was analyzed by photon counting imaging. Screened blastocysts were transferred to recipients and day 50 fetuses or calves were analyzed by PCR and Southern blot for transgene integration. When nonluminescent blastocysts were transferred, transgene integration was never observed. Of 13 fetuses derived from luminescent blastocysts, 3 contained integrated transgenes that were functional in all tissues examined. Image analysis of the signal emitted by positive blastocysts revealed that 9 nontransgenic fetuses were obtained from blastocysts that exhibited a localized luminescent signal. On the other hand, 3 of 4 fetuses derived from blastocysts that emitted light over more than 70% of their surface were transgenic. Thus, by selecting luminescent blastocysts on the basis of both signal intensity and distribution, the number of recipient females required to produce transgenic offspring can be greatly reduced. Using this technique it should also be possible to improve the efficiency of transgenesis by microinjection through studies in which vector design and integration conditions are examined at the blastocyst stage. PMID- 9859221 TI - Fertile homozygous transgenic mice expressing a functional truncated herpes simplex thymidine kinase delta TK gene. AB - Dividing cells expressing the Herpes simplex type 1 thymidine kinase (TK) can be killed upon ganciclovir treatment. Likewise, conditional cell knock-out can be obtained in transgenic mice expressing a TK gene placed under the control of tissue-specific regulatory sequences. Such animals provide powerful experimental systems for assessing the functional role of specific cell populations through their time-controlled ablation. However, whatever the regulatory sequences used, a leaky toxic overexpression of TK in testis renders male TK-transgenic mice sterile and prevents the generation of homozygous TK-expressing animals. To solve this problem, we designed a truncated TK variant (delta TK) not expressed in the testis. We generated transgenic mice expressing delta TK under the control of lymphocyte-specific regulatory sequences derived from the CD4 gene. The delta TK protein expressed in T-lymphocytes allowed the conditional ablation of activated T-cells in vitro and in vivo. Importantly, for one transgenic line we could generate fertile homozygous mice harboring a functional delta TK transgene. delta TK should thus dramatically facilitate the development of transgenic mice expressing a conditional suicide gene. PMID- 9859223 TI - Expression of allogeneic MHC class I antigens by transgenic mouse trophoblast does not interfere with the normal course of pregnancy. AB - Mammalian embryos express paternal histocompatibility antigens which make them potential targets for maternal immune responses. Yet, the histoincompatible fetus survives and develops normally. Down regulation of classical MHC antigen expression by trophoblast cells which are in direct contact with maternal circulation has been repeatedly shown. The trophoblast cells are unable to function properly in antigen presentation and do not induce allogeneic rejection reactions. In the present study we have created transgenic mice that express an allogeneic class I transgene whose transcription is controlled by the transferrin receptor promoter. The expression patterns of the transgene product mice from a single transgenic line were studied in each of the typical placental subpopulations. The allogeneic class I antigen was expressed in the allantoic plate region of the trophoblast, and this expression was not restricted to the endothelial region but extended also to the spongiotrophoblast, as well as the major blood vessels and in the endodermal sinuses. In contrast to the normal class I expression, prominent levels of allogeneic H-2 antigens were detected in the labyrinthine trophoblast. The fetal resorption rate in females mated with these transgenic males was not higher then the normal rate, and the embryos survived and developed normally. These data imply that the unusual expression of allogeneic class I antigens in certain trophoblast subpopulations does not affect fetal development. PMID- 9859224 TI - Expression of a novel piscine growth hormone gene results in growth enhancement in transgenic tilapia (Oreochromis niloticus). AB - Several lines of transgenic G1 and G2 tilapia fish (Oreochromis niloticus) have been produced following egg injection with gene constructs carrying growth hormone coding sequences of fish origin. Using a construct in which an ocean pout antifreeze promoter drives a chinook salmon growth hormone gene, dramatic growth enhancement has been demonstrated, in which the mean weight of the 7 month old G2 transgenic fish is more than three fold that of their non transgenic siblings. Somewhat surprisingly G1 fish transgenic for a construct consisting of a sockeye salmon metallothionein promoter spliced to a sockeye salmon growth hormone gene exhibited no growth enhancement, although salmon transgenic for this construct do show greatly enhanced growth. The growth enhanced transgenic lines were also strongly positive in a radio-immuno assay for the specific hormone in their serum, whereas the non growth enhanced lines were negative. Attempts to induce expression from the metallothionein promoter by exposing fish to increased levels of zinc were also unsuccessful. Homozygous transgenic fish have been produced from the ocean pout antifreeze/chinook salmon GH construct and preliminary trials suggest that their growth performance is similar to that of the hemizygous transgenics. No abnormalities were apparent in the growth enhanced fish, although minor changes to skull shape and reduced fertility were noted in some fish. There is also preliminary evidence for improved food conversion ratios when growth enhanced transgenic tilapia are compared to their non-transgenic siblings. The long term objective of this study is to produce lines of tilapia which are both growth enhanced and sterile, so offering improved strains of this important food fish for aquaculture. PMID- 9859225 TI - Expression and immunolocalisation of the snowdrop lectin, GNA in transgenic rice plants. AB - Transgenic rice (Oryza sativa L.) plants generated through particle bombardment expressed high levels of an insecticidal protein (the snowdrop lectin, GNA) directed against sap-sucking insects. Engineered plants expressed GNA either constitutively or in a tissue specific manner, depending on the nature of the promoter used to drive expression of the gene. We used specific antibodies raised against GNA to localize its expression in phloem tissue in plants engineered with the rice sucrose synthase promoter driving GNA expression. We report here molecular, biochemical and immunological analyses for fifteen independently derived transformants out of more than 200 plants we generated. PMID- 9859226 TI - Novel and transgenic food crops: overview of scientific versus public perception. AB - Recombinant DNA technology offers opportunities to develop new products in many different fields, including agriculture and the agro-food area. Transgenic plants with improved agronomic traits currently grow in field trials and a few varieties have already reached the European market. By and large, new technologies raise both concerns and expectations and modern biotechnology is no exception. Indeed, a voluntary moratorium on experiments involving recombinant DNA molecules was called for in 1974. At the present time, although a majority of academic and industrial scientists agree that transgenic food crops pose no risk for the environment or human health, some others believe that certain applications of modern plant biotechnology are hazardous. In particular, deliberate releases of genetically modified plants are regarded as risky. There is also a disparity between expert and lay perception of r-DNA technology applications to food crops, which makes public information a difficult task. This paper aims at exposing these conflicting points of view on the agricultural applications of modern biotechnology. We also propose some recommendations pertaining to public information in Europe. It appears that consensus conferences might be a good approach to stimulate public information and public debate in Europe, although this approach has to be adapted to the cultural context of each country. PMID- 9859228 TI - Chromatin heterogeneity within multicopy transgene arrays. PMID- 9859229 TI - [New techniques in heart surgery]. PMID- 9859227 TI - Efficient BLG-Cre mediated gene deletion in the mammary gland. AB - Using the phage P1-derived Cre/loxP recombination system, we have developed a strategy for efficient mammary tissue specific inactivation of floxed genes. Transgenic mice were generated which express Cre DNA-recombinase under the control of the mammary gland specific promoter of the ovine beta-lactoglobulin (BLG) gene. To test the specificity of Cre mediated recombination, we crossed these mice to animals harbouring a floxed DNA ligase I allele. We show that the BLG-Cre construct specifies mammary specific gene deletion, and furthermore that it is temporally regulated, predominantly occurring during lactation. We fully characterised the extent of gene deletion in one line (line 74). In this strain the virgin gland is characterised by low levels (7%) of Cre mediated deletion, whereas 70-80% of cells within the lactating mammary gland have undergone recombination. Immunohistochemistry and indirect in situ PCR were used respectively to demonstrate that both Cre protein and Cre activity were evenly distributed throughout the population of secretory epithelial cells. The level of background recombination in non-mammary tissues was found to be < or = 1.1%, irrespective of mammary gland developmental status. Crossing the transgenic BLG Cre strain described here to mice harbouring other floxed alleles will facilitate the functional analysis of those genes during differentiation and development of the mammary gland. PMID- 9859230 TI - [Quo vadis biomedical technology?]. PMID- 9859231 TI - [Imaging technology in medicine]. PMID- 9859232 TI - [Fluorescence--endoscopy in carcinoma of the urinary bladder]. PMID- 9859233 TI - [Minimally invasive interstitial cryosurgery with MRI guidance: possibilities for in vivo research and clinical application]. PMID- 9859234 TI - [Fluorescence examinations of cancer tissue]. PMID- 9859235 TI - [Analytic methods for functional magnetic resonance data]. PMID- 9859236 TI - [Technique for ultrasound imaging with contrast media using nonlinearity and time variance]. PMID- 9859237 TI - [Virtual neuro-endoscopy imaging of cerebral aneurysms]. PMID- 9859238 TI - [Dynamic geometric contour models for segmenting intravascular ultrasound image series in 3-dimensional surface reconstruction of coronary vessel walls]. PMID- 9859239 TI - [High resolution structural imaging of the human ear]. PMID- 9859240 TI - [Biomechanical 3D-FE model of the human cochlea]. PMID- 9859241 TI - [A 3D MLE algorithm for positron emission tomography during radiotherapy with heavy ions]. PMID- 9859242 TI - [Simultaneous control of radiotherapy with heavy ions by positron emission tomography]. PMID- 9859243 TI - [Evaluation of endoscopic images for 3-dimensional measurement of hollow biological organs]. PMID- 9859244 TI - [CT-based 3D planning for dental implantology]. PMID- 9859245 TI - [Image segmentation and 3D presentation of histological objects]. PMID- 9859246 TI - [Development of an educational program for medical ultrasound examinations: Ultra Trainer]. PMID- 9859247 TI - [Rapid neuronal filters in mammography]. PMID- 9859248 TI - [ImaView--imaging station for pre-processing tomography image data]. PMID- 9859249 TI - [Segmenting image series based on complex adaptive estimates]. PMID- 9859250 TI - [Digital data collection based on "DICOM Supplement Structured Reporting" in radiology]. PMID- 9859251 TI - [Determining cerebral circulation with electron beam computerized tomography]. PMID- 9859252 TI - [Evaluating trigger timing in detection and quantification of coronary artery calcinosis with electron beam tomography]. PMID- 9859253 TI - [Imaging cardiac impedance changes in the human thorax using impedance tomography]. PMID- 9859254 TI - [Comparison of numerically calculated surface potentials with and without considering anisotropic muscle tissue]. PMID- 9859255 TI - [Development of a 4 camera system for localizing an electrode series on the thorax]. PMID- 9859256 TI - [Optimizing electrode positions for solving the inverse problem in electrocardiography]. PMID- 9859257 TI - [Practical technical measurement problems in determining optical image errors of the human eye with a video-aberroscope]. PMID- 9859258 TI - [Determining the position of distorted point lattice patterns on the human retina by evaluating monochromatic image errors]. PMID- 9859259 TI - [3D reconstruction of anatomic structures with the TIM (tomographic imaging) visualization software]. PMID- 9859260 TI - [Development and validation of a method for wear measurement of polyethylene hip acetabulum with metal backing using roentgen images]. PMID- 9859261 TI - [Liquid dispense check]. PMID- 9859262 TI - [3D segmenting method for volume quantification of ECT (emission-computed tomography) data]. PMID- 9859263 TI - [A new measuring system for processing ultrasound multi-compression elastograph images of the prostate]. PMID- 9859265 TI - [Equipment for modification of nuclear medicine lung diagnosis as triggered SPECT -presentation, phantom studies and clinical value]. PMID- 9859264 TI - Fluorescence cystoscopy in the diagnostics and treatment of bladder tumors. PMID- 9859266 TI - [Development of a measuring system for vital microscopy]. PMID- 9859267 TI - [Development of a system for determining the position of an epiduroscope based on an optical PC mouse]. PMID- 9859268 TI - [Therapy evaluation in dermatology by comparative color measurements]. PMID- 9859269 TI - [Time-efficient and objective diagnosis of muscle function by video analysis with the BMP meter]. PMID- 9859270 TI - [Signal technique in biomedical devices]. PMID- 9859271 TI - [Determining hemodynamic status using intramyocardial electrocardiograms]. PMID- 9859272 TI - [Early detection of cardiac arrhythmias by computer-assisted online analysis of monophasic action potentials]. PMID- 9859273 TI - Applying neural network techniques to plethysmographic pulse shape analysis. PMID- 9859274 TI - [EEG analysis of the HP Component Monitoring System]. PMID- 9859275 TI - [Software platform for analysis of electro- and magnetocardiography databanks]. PMID- 9859276 TI - [Comparative assessment of isotropic and anisotropic models of signal transmission in the heart]. PMID- 9859277 TI - [Signal analysis of high resolution and intracardial electrocardiograms--use of the adaptative wavelet packet transformation for endocardial mapping]. PMID- 9859278 TI - [Multi-thread application for multi-channel signal processing and interpretation for laser Doppler flowmetry]. PMID- 9859279 TI - [Time and frequency parameters for analysis of IVUS (intravascular ultrasound) echo signals]. PMID- 9859280 TI - [Analysis of brief blood pressure and heart rate variability]. PMID- 9859281 TI - [High resolution methods for monitoring eye position]. PMID- 9859282 TI - [Technical measurement problems in a new procedure for noncontact intraocular pressure measurement]. PMID- 9859283 TI - [An eye phantom for calibrating and evaluating tonometers]. PMID- 9859284 TI - [Comparison of different evaluation methods for a multi-wavelength pulse oximeter]. PMID- 9859285 TI - [In vitro determination of substance concentration in scattering media with NIR laser remssionspectroscopy]. PMID- 9859286 TI - [The contribution of EMG for monitoring controlled electrostimulation in paralysis. 2.) Applications of EMG]. PMID- 9859287 TI - [Magnetic markers for device localization without roentgen exposure]. PMID- 9859288 TI - [Telemetry unit for transmission of performance and data from medical implants]. PMID- 9859289 TI - [Possibilities of infrared diaphanoscopy of human teeth]. PMID- 9859290 TI - [Development of an optical calibrator for commercial pulse oximeters: initial successes]. PMID- 9859291 TI - [Combined video radiography and manometry for evaluating transport function of the esophageal sphincter]. PMID- 9859292 TI - [Model studies for determining inertia of the respiratory system in ventilated newborn infants with jet impulses]. PMID- 9859293 TI - [Blood flow measurement for implantable devices. Experimental results]. PMID- 9859294 TI - [Software components for data processing of measurement values in experimental studies with PTCA catheters]. PMID- 9859295 TI - [An endotracheal tube with sensors]. PMID- 9859296 TI - [Multi-gas mainstream sensory for gas anesthesia monitoring]. PMID- 9859297 TI - [Intraoperative monitoring of the recurrent laryngeal nerve in thyroid operations]. PMID- 9859298 TI - [Electron microscopy study of coronary stents]. PMID- 9859299 TI - [Automatic EEG data processing in a multicenter study]. PMID- 9859300 TI - [A technique for imaging stretch of human back musculature with ultrasound for supporting the diagnosis of post-discectomy syndrome]. PMID- 9859301 TI - [A method for outcome control of hemodilution therapy in fetal growth retardation]. PMID- 9859302 TI - [Solution to the forward problem in electrocardiography from isotropic and anisotropic models of cardiac signal propagation]. PMID- 9859303 TI - [ECG analysis by signal pattern comparison]. PMID- 9859304 TI - [Magnetocardiography analysis of intraventricular activation by two-dimensional mapping of fragmentation of the band filtered QRS complex]. PMID- 9859305 TI - [A method for baseline correction of magnetocardiography field recordings]. PMID- 9859306 TI - [Characterization of an intravascular ultrasound measuring system for vessel wall assessment]. PMID- 9859308 TI - [Planning strategies for individual patient optimization of the dialysis process]. PMID- 9859307 TI - [Improved risk stratification by calculating QT dispersion in patients with dilated cardiomyopathy using biplanar technique in magnetocardiography]. PMID- 9859309 TI - [The "Fractionated Plasma Separation and Adsorption system" (FPSA), a new membrane absorption-assisted adjunctive extracorporeal blood purification system for liver failure]. PMID- 9859310 TI - [Microsphere based detoxification system. A new blood purification method: technical safety aspects]. PMID- 9859311 TI - [Clinical results with a new frequency-doubled, double pulse Nd:YAG laser (FREDDY) for lithotripsy in complicated choledocholithiasis]. PMID- 9859312 TI - [Suitability of modulated transillumination as coagulation control for laser induced thermotherapy]. PMID- 9859313 TI - [An automated concept for optimal ventilation of ARDS patients]. PMID- 9859314 TI - [Cell meets silicon--biomedical sensing for diagnosis and therapy]. PMID- 9859315 TI - [Micromechanical pressure sensors for medical evaluation of blood vessels and bypasses after surgical intervention]. PMID- 9859316 TI - [Technical microsystem monitoring for determination of metabolic parameters]. PMID- 9859317 TI - [Intraoperative monitoring of a robot-assisted minimal invasive mitral valve solo surgery]. PMID- 9859318 TI - [Development of a modular robot system for microsurgery]. PMID- 9859319 TI - [Development and characterization of a modified external fixator for inducing and monitoring micro-movement in fracture zones]. PMID- 9859320 TI - [Development and evaluation of a diagnostic treadmill for three-dimensional imaging of gait dynamics]. PMID- 9859321 TI - [Tensile strength of carbon fiber reinforced implant posts in quasi-statistical and increasing stress using simulated in vivo conditions]. PMID- 9859322 TI - [Analysis of tension distribution in bone matrix on a stressed external fixator bone pin in a two-dimensional model with finite element method of tension optics]. PMID- 9859323 TI - [Construction of a multiaxial testing system for connecting elements of modular endoprostheses]. PMID- 9859324 TI - [Refixation of the patellar ligament with special tumor endoprostheses]. PMID- 9859325 TI - [Development of a cone tapered connection for modular prosthesis systems of carbon fiber compounds]. PMID- 9859326 TI - [Comparison of national and international standards of electrical safety of medical devices]. PMID- 9859327 TI - [Electromagnetic interference with implants]. PMID- 9859328 TI - [Effect of mobile phones on medical devices]. PMID- 9859329 TI - [Telecardiology--a service performance option for the physician]. PMID- 9859330 TI - [Adaptive drift detection and intelligent alarm systems]. PMID- 9859331 TI - [Quality assurance of Doppler color ultrasound equipment]. PMID- 9859332 TI - [Recommendation for measuring magnetic interference according to the DIN 570107 A1 norm]. PMID- 9859333 TI - [Post-war studies of health risk caused by excessive military microwave exposure. What can these studies contribute to current risk assessment of electromagnetic fields?]. PMID- 9859334 TI - [Modeling and analysis of extensive electric sources: the magnetic field of stimulated peripheral nerves]. PMID- 9859335 TI - [Modeling the magnetoencephalogram from extended sources]. PMID- 9859336 TI - [Reconstruction of extended electric sources in a thoracic phantom using biomagnetic field measurements]. PMID- 9859337 TI - [Detecting the spatial distribution of QT dispersion in bipolar magnetocardiography with various myocardial infarct sites]. PMID- 9859338 TI - [Measuring para-DC biomagnetic fields of the head using a horizontal modulated patient cot]. PMID- 9859339 TI - [Results of ECG analysis by signal pattern comparison]. PMID- 9859340 TI - [Risk stratification by analysis of QT dispersion and late potentials in the electrocardiogram and biplanar magnetocardiogram in patients with coronary heart disease]. PMID- 9859342 TI - High-resolution ERD using realistic head geometry. PMID- 9859341 TI - [Fetal sinus bradycardia in the magnetocardiogram as an expression of sympatho vagal imbalance in the 2nd trimester]. PMID- 9859343 TI - [Contribution of EMG to monitoring controlled electrostimulation in paralysis: 1) Realization of a modular EMG recording hardware]. PMID- 9859344 TI - [Use of impedance spectroscopy in biotechnology]. PMID- 9859345 TI - [Topologic distribution of slow EEG components using various stimulation paradigms]. PMID- 9859346 TI - P-wave analysis in MCG and ECG after conversion of atrial fibrillation. PMID- 9859347 TI - [MEG studies of the somatosensory system by passive hand movement]. PMID- 9859348 TI - [Technical realization of a neural feedback system]. PMID- 9859349 TI - [Event-related analysis of interval sequences]. PMID- 9859350 TI - [In vitro evaluation of an implantable measuring system for impedance spectroscopy for evaluating tissue status]. PMID- 9859351 TI - [Noninvasive diagnosis of joint damage using an acoustic emission analysis]. PMID- 9859352 TI - [Optimizing a common axis-controlled knee joint prosthesis for tumor patients]. PMID- 9859353 TI - [Verification of an ultrasound measuring procedure for evaluating screw-bone fusion based on mechanical parameters]. PMID- 9859354 TI - [Exhalation monitoring with online analysis equipment--ppb and sub-ppb analysis with a improved sensors]. PMID- 9859355 TI - Human tracking performance under multitasking conditions. PMID- 9859356 TI - [Determination of group classification of Raynaud patients by discriminance analysis of laser Doppler signals]. PMID- 9859357 TI - [Active shielding for improved interference field suppression of magnetically shielded chambers]. PMID- 9859358 TI - [Device for three-dimensional measuring and frequency analysis of electric and magnetic fields]. PMID- 9859359 TI - [Micromechanical drug applicator for ophthalmology]. PMID- 9859360 TI - [Interactive computer program for separating respiratory pressure in newborn infants into its elastic, viscous and inert components]. PMID- 9859361 TI - [Experimental development of non-dispersive measurement of 13C/12C isotope relations in gases]. PMID- 9859362 TI - [Medical man-machine systems]. PMID- 9859363 TI - [Model development of biological processes exemplified by intracranial pressure]. PMID- 9859364 TI - [Calculation of the flow through a new single cusp valve for heart assist systems]. PMID- 9859365 TI - [Simulation of deglutition for quantitative study of micro-aspiration in mechanical ventilation]. PMID- 9859366 TI - [Calculation of asymmetric temperature fields in hyperthermia treatment with isolated extremity perfusion]. PMID- 9859368 TI - [Monophasic action potential over long-term follow-up]. PMID- 9859367 TI - [Complexity of PP intervals in a nonlinear model of patients after heart surgery intervention]. PMID- 9859369 TI - [An integrated data model in biomedicine]. PMID- 9859370 TI - [A model for simulating heat transfer and thermoregulation of premature infants]. PMID- 9859371 TI - [A multifunctional coronary circulation model: various applications for comparative studies of PTCA catheters]. PMID- 9859372 TI - [Selective, vibration-induced destruction of malignant cells]. PMID- 9859374 TI - [Detection of partial obstructions of the endotracheal tube]. PMID- 9859373 TI - [Follow-up of acoustic respiratory impedance in simulated obstruction of the upper airways]. PMID- 9859375 TI - [Modeling of the baroreceptor reflex in a pulsatile model]. PMID- 9859376 TI - [Flow dynamics of an over-dimensional end-to-side anastomosis in a circulatory model with numerical flow simulation]. PMID- 9859377 TI - A Simulink model for the human circulatory system. AB - This paper presents the mathematical simulation of the human circulatory system. The model is based on the former work of Coleman and co-workers and has been redesigned for simulation with the Matlab toolbox "Simulink". It includes the heart and the peripheral circulation, the respiratory system, the kidneys and the major neural and hormonal control mechanisms, which are necessary for maintaining homeostasis. The model contains more than 30 blocks with over 200 physiological variables, which can be accessed and plotted during the simulation. PMID- 9859378 TI - [Separating elastic, flow and velocity work and work of smooth musculature of the peripheral arterial pulse of the human]. PMID- 9859380 TI - [Modeling intracardiac potentials using the finite element method]. PMID- 9859379 TI - [Coupling cardiovascular models for analysis of frequency-adaptive cardiac pacemaker systems]. PMID- 9859381 TI - ["Nonlinear differential equation for assessing the stability of heart muscle excitation"]. PMID- 9859382 TI - [Modeling support element forms in quasi-telescopic external fixators]. PMID- 9859383 TI - [Regulation of the heart-lung machine: systems analysis]. PMID- 9859384 TI - [Field parameter studies of hollow fiber membrane oxygenator systems for extracorporeal blood circulation]. PMID- 9859385 TI - [Comparative studies of hemolysis in blood pumps of various designs]. PMID- 9859386 TI - [Development of a predictive MIMO regulator for heart-lung machines based on a model of extracorporeal circulation]. PMID- 9859388 TI - [Animal experiment for testing the application of perfluorocarbon-assisted membrane oxygenation]. PMID- 9859387 TI - [Procedure for membrane oxygenation of blood over hydrophilic polymer membranes]. PMID- 9859389 TI - [Assistance systems for minimal invasive surgery]. PMID- 9859390 TI - [Pneumatic external counterpulsation--a new technical perfusion principle]. PMID- 9859391 TI - [Expert system for knowledge-based Holter data analysis]. PMID- 9859392 TI - [Long-term measurement of unipolar intracardiac impedance with an implantable cardiac pacemaker and Holter monitor]. PMID- 9859393 TI - [Control algorithm for the "dromotropic" cardiac pacemaker concept]. PMID- 9859394 TI - [A technical regulating approach to dromotropic control of closed-loop frequency adjustment]. PMID- 9859395 TI - [Effect of tissue impedance on the ECG and MCG signal: a phantom study]. PMID- 9859396 TI - [Atrial fibrillation--study of electrophysiologic influences on optimizing cardiac stimulation methods]. PMID- 9859397 TI - [Relation of defibrillation threshold on shock duration and discharge capacity. Determination of chronoaxis-rheobase relations of defibrillators]. PMID- 9859398 TI - [Time-dependent balloon expansion of PTCA catheters]. PMID- 9859399 TI - [Reaction force studies in advancement and retrieval of PTCA catheters in coronary vessel models]. PMID- 9859400 TI - [Quantifying stenosis and evaluating projection--an online realization in the catheter laboratory]. PMID- 9859401 TI - [Foreshortened imaging: a new method for spatial interpretation and evaluating the view of the coronary region of interest]. PMID- 9859402 TI - [3-dimensional activity analysis with activity sensors for pacemaker therapy]. PMID- 9859404 TI - [Measuring and correlation of microbubbles in extracorporeal circulation]. PMID- 9859405 TI - [Optimizing 2-electrode arrays of implantable defibrillators]. PMID- 9859403 TI - [1-year follow-up of traumatic pacemaker electrodes]. PMID- 9859406 TI - [Interaction of radial strength and flexibility of coronary stents]. PMID- 9859407 TI - [Effect of substance and length of intramedullary implant rods of special tumor prostheses on bone canal]. PMID- 9859408 TI - [Effect of bioactive coats of natural and synthetic hydroxyapatite on repair and induction of bone tissue processes]. PMID- 9859409 TI - [Characterization of CVD diamond-coated titanium base compounds for biomedical applications]. PMID- 9859410 TI - [An in vitro study of hydrodynamic properties of the TRI-Technologies heart valve prosthesis in accordance with prEN 12006-1]. PMID- 9859411 TI - [Labcor-stented porcine heart valve prosthesis in laboratory testing in accordance with prEN 12006-1]. PMID- 9859412 TI - [Effect of electronic surface properties on interaction between fibrinogen and solid bodies]. PMID- 9859413 TI - [Correlation of physicochemical surface properties of various synthetic granulates with their hemocompatibility]. PMID- 9859414 TI - [In vitro studies of hemocompatibility of surface modified fluoropolymers]. PMID- 9859416 TI - [Evaluation of biomaterial surfaces by detection of mechanical and electrical inhomogeneities using scanning probe microscopy]. PMID- 9859417 TI - [Micromorphometry and stereography of rough depth relief coated implant surfaces by scanning electron microscopy and confocal laser scanning microscopy]. PMID- 9859415 TI - [Techniques for determination of the microstructure and elastomechanical properties of cortical bone]. PMID- 9859418 TI - [Different effects of various embedding techniques on confocal laser scanning microscopy image and mechanical properties of cortical bone]. PMID- 9859419 TI - [Bioactive coats on titanium implants and biocompatibility of titanium compounds for orthopedic implants]. PMID- 9859420 TI - [Porous materials for reconstruction of bone tissue]. PMID- 9859421 TI - [Collagen bionation of silicon, as a material for a lens epithelium coated capsular sack model]. PMID- 9859422 TI - [Substance-technical and optical studies of test bodies of hydroxylapatite ceramics]. PMID- 9859423 TI - [Innovative use of polymer substances in medical technology--form stability and surface modification]. PMID- 9859424 TI - [Sound transmission properties of synthetic prostheses in ossicular replacement during middle ear reconstruction]. PMID- 9859425 TI - [Design of a test chamber for simulation of adhesion pressure of the anterior lens capsule on the intraocular lens]. PMID- 9859426 TI - [Development of a multifunctional cell chamber for vital microscopy]. PMID- 9859427 TI - [Registration of hardness measurements of biomaterials in the nanometer area with scanning force microscopy studies]. PMID- 9859428 TI - [Roentgen contrasting of resorbable biomaterials PHB and PLA]. PMID- 9859429 TI - [Comparative studies of the degradation kinetics and effect on mechanical properties of polylactide and polyhydroxybutyric acid in vitro]. PMID- 9859430 TI - [Effect of softening agents on biodegradation and biocompatibility of poly-beta hydroxybutyric acid in vivo]. PMID- 9859431 TI - [The hemostasis system as an indicator of hemocompatibility of implant materials]. PMID- 9859432 TI - [Does heparin coating improve the biocompatibility poly(D,L)lactide?]. PMID- 9859433 TI - [Mechanical properties of metal vascular implants (stents)]. PMID- 9859434 TI - [Biocompatibility of glass and silicon]. PMID- 9859435 TI - [Cytometry and cell sorting]. PMID- 9859437 TI - [A new method for ex vivo cell and tissue culturing]. PMID- 9859436 TI - [Cellular engineering: crash tests with human erythrocytes reveal hidden properties of cellular proteins]. PMID- 9859438 TI - [Optical measuring methods for monitoring and evaluation of experimental cell and tissue cultures in undisturbed and flowing media]. PMID- 9859439 TI - Quantitative laser microscopy in single cells based on ratio-imaging of fluorescent indicators: a critical assessment. PMID- 9859440 TI - [MMD-NEURODOC: Multimedia Medical Database--electronic medical library]. PMID- 9859441 TI - [Modular studying--the new concept of medical technology curriculum of the Ulm Technical University]. PMID- 9859442 TI - [Education in biomedical technique: past--why--future?]. PMID- 9859443 TI - [New aspects of "electro-technology" in the engineering curriculum at the Dresden Technical University]. PMID- 9859444 TI - [Technical perfusion curriculum--an additional curriculum and complementary graduate curriculum in biomedical device technique]. PMID- 9859445 TI - [Concept of the "ECG signal analysis" teaching trial]. PMID- 9859446 TI - [Calibration of volume data from spiral CT images for defining the contour of individually fitted skull implants]. PMID- 9859447 TI - [Effect of corrective procedures on volume quantification of SPECT data]. PMID- 9859448 TI - [Segmentation-based movement estimation in ultrasound images]. PMID- 9859449 TI - [3D localization of electrodes on and in the human body based on biplanar roentgen images]. PMID- 9859451 TI - [Compression of 3- and 4-dimensional medical image data]. PMID- 9859450 TI - [Local resolution in temperature measurement using nuclear magnetic resonance tomography]. PMID- 9859453 TI - [Teaching materials from image data of the human body]. PMID- 9859452 TI - [Optical laboratory tomograph for education and research]. PMID- 9859454 TI - [An analytic model for blood pressure transmission distance of invasive blood pressure determination with external liquid coupling]. PMID- 9859455 TI - [Controlled calibration site for sensors for invasive blood pressure measurement]. PMID- 9859456 TI - [Vector magnetometer module for biomagnetic measurements]. PMID- 9859457 TI - [Cellular automated detection for simulation of the impulse transmission with reference to anisotropy]. PMID- 9859458 TI - [Evaluation of experimental cell and tissue cultures for improved tissue culture monitoring]. PMID- 9859459 TI - [Ellipsometric study of adsorption behavior of blood proteins fibrinogen and serum albumin to solid body surfaces]. PMID- 9859460 TI - [Cytotoxicity and scanning electron microscopy studies for determining biocompatible of solid body surfaces]. PMID- 9859461 TI - [Effect of iatrogenic damage to intramedullary nails]. PMID- 9859463 TI - [Pulsatile flow model of the heart for validating new echocardiography methods for determining regurgitation volume]. PMID- 9859464 TI - [Interference behavior of DDD pacemakers]. PMID- 9859462 TI - [An improved cardiovascular model for normothermic organ perfusion]. PMID- 9859465 TI - [Expansion of the initial depolarized tissue zone in cardiac pacemaker stimulation]. PMID- 9859466 TI - [Symbolic dynamics--an independent method for detecting nonlinear phenomena of heart rate regulation]. PMID- 9859467 TI - [Experimental study of the radial deformability of stents]. PMID- 9859468 TI - [A new heart valve for heart assist systems]. PMID- 9859469 TI - [Mechanical properties of human and swine pericardium]. PMID- 9859470 TI - [Method for calibrating spiral CT data in contour finding processes in individual fitting of skull implants]. PMID- 9859471 TI - [3-dimensional ultrasound elastography of the female breast: in vitro and in vivo results]. PMID- 9859472 TI - [A system for detection and imaging geometric values from video images for movement analysis of the human body]. PMID- 9859473 TI - [Calibration of roentgen image intensification in computer-assisted orthopedic surgery]. PMID- 9859475 TI - [Preprocessing of the Visible Female data]. PMID- 9859474 TI - [Segmentation of clinical 4-dimensional magnetic resonance tomography images using active contour models and haptic interaction]. PMID- 9859476 TI - [Electromagnetic field propagation in tissues to characterize channel properties for implant telemetry]. PMID- 9859477 TI - [Dual sequence method for analysis of spontaneous baroreceptor sensitivity]. PMID- 9859478 TI - [Measuring bioimpedance in dialysis patients for determining the amount of water in individual fluid compartments of the body]. PMID- 9859479 TI - [Validation of a robot for determination of regional perfusion with fluorescent microspheres]. PMID- 9859480 TI - [Validation of a filtration vessel to determine regional blood flow of the heart using fluorescent microspheres]. PMID- 9859481 TI - [Supply of disability aids for handicapped patients]. PMID- 9859482 TI - [Bioresorbable screws in comparison with titanium screws for osteosynthesis after sagittal fracture of the mandible--a prospective, randomized, controlled clinical study]. PMID- 9859483 TI - [Design of a gavanically isolated wide SCSI processor equipment interface for bidirectional data transfer with a data rate of 10 MByte/s]. PMID- 9859484 TI - [Microsystem technique for medical applications: overview of combined projects and prospects]. PMID- 9859485 TI - Implantable telemetric endosystem for biomedical applications--concept and realization. PMID- 9859486 TI - Telemetric transmission of energy and data for implantable systems. PMID- 9859487 TI - [Development of a miniaturized system for ambulatory home use in initial diagnosis of sleep apnea disorders]. PMID- 9859488 TI - [Highly integrated microsystem for neurostimulation]. PMID- 9859489 TI - [Development of a surface coating for spinal cord stimulation]. PMID- 9859490 TI - [Fiber optic dosimeter for radiotherapy]. PMID- 9859491 TI - [Fully implantable in vivo microdialysis system for glucose monitoring. Total system concept and implantable prototype system]. PMID- 9859493 TI - [Miniaturized cardioverter defibrillator]. PMID- 9859492 TI - [Evaluation of VISY-1 in an in vivo test: sampling and glucose determination]. PMID- 9859494 TI - [Micro-optic tonometer]. PMID- 9859495 TI - [Development of an analytic standard for evaluating heart rate variability in cardiovascular autonomic neuropathy in diabetes: comparison of short-term spectral analysis with Ewing's standard battery of reflex tests as reference method]. PMID- 9859496 TI - [University research and education in rehabilitation technology: inducing ISO 9001 conformity and initial experiences with a certified quality management system]. PMID- 9859497 TI - [Vital signs information representation--on the way to a standard for communication of medical devices in anesthesia and intensive care medicine; 2: Nomenclature and data dictionary]. PMID- 9859498 TI - [Laser Doppler flowmetry for microcirculation monitoring. A concerted European action]. PMID- 9859499 TI - Platform independent implementation of DICOM-based miniPACS. AB - Platform independence is an important issue for a software designed to run across a hospital environment which is practically always heterogeneous one. This paper presents a simple miniPACS solution designed over an existing Intranet (Internet limited to a local area network). Platform independence is achieved with Java programming language which allows resulting code to be executed on virtually all machines. System consists of a server application providing DICOM (Digital Imaging and Communications in Medicine) services and clients that can use these services over a TCP/IP connection. The server application is capable of an image storage and supports a query mechanism to allow access to stored data, thus the clients may perform two major functions: providing image data for archiving or image retrieval for viewing. Presented client application provides both these functions. It retrieves stored images for further processing and returns calculated results to the archive. The parts of the system are connected through a DICOM interface. Consequently the system architecture is open and any devices like scanners or viewing stations supporting specific DICOM services may be attached. PMID- 9859500 TI - [Containment sensors: manufacture in technical production facilities--technical and economic aspects]. PMID- 9859505 TI - [Prevalence of iron deficiency in early infancy]. AB - In order to determine the prevalence of iron deficiency among 6-to-24 month-old children in the Cascais county, a cross-sectional study with systematic opportunist sampling of all 6-to-24-month-old children seen for routine immunization or well-child health care at the Public Health Centers in the Cascais county, during the Spring of 1994. All children were submitted to a short nutritional and clinical inquiry. Blood samples to evaluate anemia and iron deficiency were obtained from eligible children after parental consent. Of the 183 children who visited the health centers during the enrollment period, 125 were eligible for blood sampling. Adequate blood samples were obtained from 120; 38 (31.7%) fulfilled the eligibility criteria for the therapeutical trial (polymaltose-ferric hydroxide, 5 mg/kg/day, for 4 weeks). Twenty-six completed the trial and 13 (50%) had a positive response. The prevalence of iron deficiency estimated for this population sample was 15.8% (CI95% = 8.8-22.3), twice the prevalence of iron deficient anemia, presenting only as mild anemia. Nevertheless, for a total population of 3,500 6-24 month old children in the Cascais area, a 15% prevalence means over 500 iron-deficient children (estimated range between 300 and 800) of which 250 are likely to suffer anemia (150-400) and may be at risk of developing a permanent intellectual deficit. The size of the problem as sampled in the Cascais area does not justify recommending the screening of all young children; instead it points to the need to pay special attention to nutritional education in well-child health care visits, and to allow the early detection and treatment of infants with iron deficiency or at risk of becoming iron deficient. PMID- 9859507 TI - [Pediatric varicocele]. AB - The treatment of pediatric varicocele has recently been given a special importance due to clinical and experimental evidence of testicular atrophy and histological changes in 50% of homolateral and sometimes in contralateral testis. Those changes are progressive and similar to those in infertile adults with varicocele. Between 1st January 1990 and 31st December 1994, 41 boys (five to 15 years old--mean age 11.9 years) were presented for evaluation of primary varicocele at the Pediatric Surgery Department of Dona Estefania Hospital. The varicocele was left sided in 39 boys, bilateral in one and another on the right, with a predominance of grade II. Forty patients underwent varicocelectomy, 31 (77%) by the Ivanisevic technique. Varicocele recurred in four cases (10%). The authors discuss the results and make some considerations concerning the management of varicocele in children. PMID- 9859506 TI - [Importance of J. Brazy's neurobiological index. Prediction of the number and severity of complications in very low birth weight infants]. AB - The objective of this study was to evaluate the neurodevelopment outcome of Very Low-Birth-Weight Infants (VLBW), between 1987 and 1993, and correlate these findings with J.Brazy's Neurobiologic Risk Score (NBRS). The minimum corrected age at follow-up was 12 months. The neurodevelopmental assessment was performed using Mary Sheridan and Ruth Griffiths scales, Auditory and Visual Brainstem Evoked Responses and Stycar test. The NBRS was applied to 77 children. According to the score, three groups of risk were defined: Low < or = 4; Intermediate 5-7; High > or = 8. We obtained the following results: children with NBRS < 4, 20% had handicaps (5% of which major); children with NBRS 5-7, 41% had handicaps (23% of which major); in children with NBRS > 8, 95% had handicaps (80% of which major). The incidence of handicaps, (all grades included) was 71.4% for those weighing less than 1000 gr at birth, and 39.2% for those weighing 1000-1499 gr at birth. Major handicaps, mainly motor deficits, occurred in 26.8% of VLBW infants and minor to moderate handicaps were observed in 18.3% of patients in this group. These results were compared to J.Brazy's originals. We concluded that the NBRS, which is simple and objective to perform, is a good predictor of subsquent abnormal development in VLBW infants, allowing the infant's integration as soon as possible in high-risk follow-up programs, to place as soon as possible. PMID- 9859508 TI - [Prepubertal testicular tumors]. AB - Prepubertal testis tumours (T.T) are rare, with different characteristics and clinical course to adults and with a better prognosis. The authors report the experience of Maria Pia Hospital's over a 15 year period. There were 8 cases of T.T.: 3 Yolk sac tumours, 3 teratomas, 1 Leydig cell tumour and 1 epidermoide cyst. All patients are submitted to an inguinal orchiectomy. Follow-up ranged between six months and 14 years. There was no recurrence of the disease in seven patients are without disease recurrence. Mortality was nil. PMID- 9859509 TI - [Nephrotic syndrome. What is new since the 1988 study?]. AB - The authors make a retrospective review of 53 new cases of Nephrotic Syndrome followed up in the Nephrology Unit from November 1988 to March 1994, bearing in mind the evaluation of casual changes of the disease standard regarding a previous study of 1988. Epidemiological, clinical, therapeutical and evolutional aspects were studied. Forty-four cases of primary Nephrotic Syndrome (83%) were identified, 61.4% of which behaved as cortico-sensitive, 25% as cortico dependent, and 13.6% as cortico-resistant; 8 cases (15%) of Nephrotic Syndrome secondary to infection, Systemic Lupus Erythematosus and Amyloidosis, and 1 case of congenital Nephrotic Syndrome (2%). The theory that the high number of cortico dependent is, probably, related with a higher severity in the relapse diagnosis and/or changes in the children's standard of living is admissible. It was also observed that at present there is a lower number of hospital discharges, related to more careful attitudes adopted regarding the evolution of the disease. PMID- 9859510 TI - [Bladder malfunction, urinary tract infection and vesicoureteral reflux in children]. AB - A correlation between urinary tract infection, vesicoureteral reflux and voiding disorders has increasingly been reported. Voiding dysfunction increases the incidence of recurrent urinary tract infection, induces and perpetuates vesicoureteral reflux, even after surgical antireflux treatment, and may result in permanent renal damage. The resolution of the primary cause with voiding normalization is essential to achieve good results in the treatment of secondary problems such as urinary tract infection and vesicoureteral reflux. Thirty seven children with vesicoureteral reflux secondary to voiding disorders were diagnosed and treated between 1990 and 1995 (five years). Forty-nine ureters were studied. The subjects became symptomatic between 1 month and 13 years of age, with the occurrence of urinary tract infection. All children were neurologically and morphologically normal. Symptoms suggesting bladder instability were detected in 34 (91.9%) and dysfunctional sphincter obstruction in three (8.1%). These patients were all evaluated with a renal/bladder sonogram and voiding cystogram, complemented in 17 (45.9%) with urodynamic testing that confirmed clinical diagnosis. 99mTc-dimercaptosuccinic acid renal scans performed on 29 (78.4%) children revealed renal damage in 26 (89.6%). A treatment program of bladder retraining and bowel habit normalization was encouraged in every child, anti cholinergic drugs were associated in 23 (62.2%), muscle-relaxant drugs in three (8.1%), phenoxybenzamine and intermittent catheterization were used in one child (2.7%). Urinary tract infection prophylaxis was instituted in 34 (91.9%) children. Urinary tract infection was completely resolved in 35 (94.6%) patients, and its frequency decreased in two (5.4%). Thirty-two children (86.5%) with vesicoureteral reflux were cured and four (10.8%) were improved. Evidence of voiding disfunction ceased in 22 (59.5%) cases and improved in 14 (37.8%) with a reduction in the frequency and intensity of complaints. Urgency syndrome and vesicoureteral reflux remained unchanged in one child (2.7%). These findings imply that detection and treatment of bladder/sphincter disfunction are essential in every child with the complex of recurrent urinary tract infection and vesicoureteral reflux. PMID- 9859512 TI - [Provision of pediatric cardiology care in Macau. Initial experience of the central hospital of Conde de S. Januario]. AB - In the last decades, the conceptual and technical advances in paediatric cardiology have brought additional options concerning the provision of paediatric cardiology care. The purpose of this study was to analyse the initial experience of a paediatric cardiology service in a conventional general hospital serving a population of nearly 450,000 inhabitants, mostly Chinese. Access to several levels of care, including cooperation with a tertiary referral center, is described. From September 1993 to July 1995, 717 children with either suspected or diagnosed heart disease were seen: 294 (41%) were discharged and, of the remaining 423 with heart disease, 364 (85%) had congenital heart disease; 75% were adequately managed with the available local resources. Appropriate referrals provided more advanced approaches for the smaller group requiring them. The training of local doctors has ensured the continuity of a good standard of care for this group of paediatric patients in Macau, with an evident reduction in direct as well as indirect costs. PMID- 9859511 TI - [Acute mastoiditis in children]. AB - We retrospectively studied seventeen cases of acute mastoiditis admitted to the Paediatric Ward of S. Francisco Xavier Hospital during 8 years and 8 months. Seven patients were admitted in 1995. We found no reason for this increase. Eleven children were male and 5 were under 2 years of age. All cases, except one of post-traumatic mastoiditis, occurred after acute otitis media. Only 5 children referred symptoms for more than seven days before admission. Seven patients had surgery. The surgical group of children had a longer period of illness when compared to those only treated medically (7.6 versus 3.9 days). Tympanocentesis was performed in 6 patients, but it did not affect the outcome of the illness. In three children there were complications: Bezold abscess, labyrinthitis and cholesteatoma. The latter was the only patient in our series with permanent hearing loss. PMID- 9859513 TI - [Prenatal cardiology. Suspicion to confirmation]. AB - The mortality rate is high and prognosis is worse among new-borns with prenatal diagnosis of heart malformation, mainly due to factors such as its association with other malformations, and a range of more severe diseases probably resulting from the predominance of the obstetric use of the four chamber view. In this study we retrospectively assessed the range of cardiopathies diagnosed by foetal echocardiography and their evolution, compared with previous years. From January 1994 to December 1995, 1173 foetal echocardiograms were performed at a gestation age of 24 weeks. Sixty-one foetuses (5.2%) had cardiac anomalies, structural in 56 and arrhythmia in 5. The risks and indications were maternal in 37%, foetal in 31%, familial in 17% and environmental in 15%. Three were false negatives (VSD:2; truncus arteriosus: 1). Five died in utero, and 18 were assessed after birth with a mean gestational age of 37 weeks and birth weight of 3 Kg, a caesarean section was performed in 9. All but one were born in central hospitals. Six children were operated on. Two children died, one after surgery. Compared with the four previous years of activity, indication due to foetal risk rose from 6 to 31%, the number of cases diagnosed with heart disease increased from 14 to 30 per year, and the mortality decreased from 59 to 11%. Despite this, we still observe that the vast majority of new-borns who are hospitalised due to a severe heart disease had no prenatal diagnosis, indicating the need to continue our educational policy in this field. PMID- 9859515 TI - [Free me from this nail! Please!]. AB - Ingrowing toenails are a very frequent pathology in children and adolescents. There is an anatomic component, but also other factors, such as inadequate shoes and incorrect nail-trimming. We treated 47 patients with 69 ingrowing toenails, between one and 15 years of age. The evolution time was one month to 10 years and 11 patients had precedent surgeries. Lateral electrosurgical matricectomy was the choice technique. With a follow up of at least eight months, there is only one case of recurrence. PMID- 9859514 TI - [Branched chain amino acid diseases]. AB - The authors present 19 cases of branched-chain AA catabolism disease: 9 Maple Syrup Urine Diseases, 6 Methylmalonic Acidemias, 2 Propionic Acidemias, 1 case of 3-OH-3-methylglutaryl-CoA-lyase deficiency and another of 2-methyl-ketoacetyl-CoA thiolase deficiency. Fifteen are early neonatal forms and in 4 the onset occurred later. Fifteen patients (78.9%) needed one or several extra-corporal procedures either in the initial acute phase or during relapse. Fifteen patients presented several metabolic relapses, sometimes fatal (3 children). Global mortality was 26.3%, that is 5/19 patients: 4 children with neonatal forms, one with a later onset. Eleven patients (57.9%) had an IQ/DQ > or = 80: only 46.6% of the neonatal forms obtained these results, in contrast with all the late onset forms. Of the survivors, 9 (64.2%), presented a residual neurologic condition. The correct diagnostic and treatment procedures are defined and the need to consider the existence of these diseases at any age, particularly in the neonatal period, is stressed. PMID- 9859516 TI - [Clinical and laboratory basis for the diagnosis and understanding of neonatal metabolic diseases]. AB - The authors divide neonatal metabolic diseases into two major groups: intoxication and energy deficiency. The main signs which allow for the suspicion of the diagnosis are indicated for each group. The complementary examinations to be carried out by the Clinical Pathology Service of the Central Hospital and those which must be carried out by the metabolic diseases Reference Centre are reviewed. Based on the clinical framework and on the examination results, the authors establish five syndromatic groups to orientate diagnosis. The authors conclude by presenting differential diagnosis tables based on the original systematic classification by Jean-Marie Saudubray, with up-dated modifications from their own experience. PMID- 9859517 TI - [Lipid disorders in children with insulin-dependent diabetes mellitus]. AB - Lipid and lipoprotein disorders are frequently detected in insulin dependent diabetics, which predisposes the high cardiovascular risk present in these patients. Studies performed with insulin dependent children showed early changes in lipid metabolism, usually correlated and aggravated by poor glycemic control. However, there are abnormalities present even in diabetic children with good glycemic control. The usual measures used to improve diabetic control are not sufficient to correct all the lipid disorders in Diabetes Mellitus. Hyperglycemia is the major factor, inducing metabolic lipid changes by increasing hepatic synthesis of triglycerides and promoting lipoprotein and apolipoprotein glycosylation and oxidation. Other changes, associated with the decrease of lipoprotein lipase activity are directly related to insulin deficiency. The lipid profile in children with poor diabetic control is similar to that already described for adult patients. The main abnormalities found are: increased levels of triglycerides, VLDL-Tg, LDL-Tg, VLDL-Cholesterol, Apo B and Apo CIII, with decreased values of HDL-Cholesterol and Apo AI. As there is a strong correlation between control and the degree of lipid changes, even with normal levels of cholesterol and triglycerides, measurements of Apo AI, Apo B100 and Apo CIII seem to be good and reliable indicators of glycemic control in diabetic children, and a factor with high predictive value for the evaluation of cardiovascular risk in adult patients. PMID- 9859518 TI - [Latent AIDS in young children of mothers who were HIV-negative in the first trimester of pregnancy]. AB - The authors describe two cases of infants admitted to our Unit with respiratory failure caused by a Pneumocystis Carinii pneumonia (PCP). Because their mothers were HIV negative in the first three months of pregnancy. HIV infection was not suspected and the diagnosis was only made after the PCP. Based on these examples we discuss the importance of detecting HIV infection during pregnancy. PMID- 9859519 TI - [Congenital cyst adenomatoid malformation of the lung]. AB - The authors report two cases of Congenital Cyst Adenomatoid Malformation diagnosed by ultrasound in the 24th and 27th weeks of pregnancy. The pregnancies had no other problems until their term and deliveries were normal. Surgical resection of the malformation was made on the 8th day of life in one child and on the 11th day in the other. After 2 years, the first child shows a good evolution in weight and height, without lung pathology. After 2 months, the other child also shows a similar condition. PMID- 9859520 TI - [Extra-lobar pulmonary sequestration with broncho-esophageal fistula]. AB - We describe a case of extra-lobar pulmonary sequestration with broncho-esophageal fistula in a newborn male who presented respiratory distress, cyanosis and feeding difficulties. The diagnosis was made with a swallowed contrast examination, nuclear magnetic resonance, digital subtraction angiography and confirmed by gross and histologic examination. The work of a multidisciplinary team was essential for an early diagnosis and the correct and effective treatment of this Cuncommon condition. PMID- 9859521 TI - [Pediatric Crohn's disease complications]. AB - This case report describes our experience in treatment of a child with Crohn's disease involvement of ileocecal and sigmoid regions complicated with a severe abdominal wall abscess. A combined therapeutic approach of the acute episode was based on intestinal diversion, abscess drainage, antibiotherapy, nutritional support and post-operative immunosuppression with azathioprine. The definitive treatment of the ileocolonic involvement occurred 6 months after the acute episode and consisted of limited resection, multiple strictureplasties, primar" closure of ileosigmod fistula and undiversion. This operative procedure was followed by a short period of total parenteral nutrition and progressive re establishment ofenteric nutritional support and immunosuppression. At 18 months follow-up, under low dose azathioprine revealed, a normal growth with a 20 Kg weight gain and 10 cm height gain, no evidence of fistula recurrence or restenosis and suggests the safety of strictureplasty in paediatric patients as a way to conserve as much small bowel as possible. PMID- 9859522 TI - [A breech delivery and induction with prostaglandin E2]. AB - Induction of labour with local application of PgE2 in breech presentation is still a much discussed problem. We aimed at studying the effect of local application of PgE2 (Prostin E2--3 mg) vaginal tablets for induction of labour in breech presentation. 15 pregnancies were studied and induced--8 for postdate, 6 for prae-eclampsia and 1 for fetal demise. Sonographic biometry and cardiotocographic examinations were conducted as well as a precise bishop score. Labour began within the interval of 3-13 hours after the application on the first tablet. The length of delivery was 4-24 hours. Effective labour was instituted in 93.33% on the parturiens, while in 1 (6.67%) there was a need for augmentation with oxytocin. There were no cases of hyperstimulation. Vaginal delivery was accomplished in 100% of the patients among which the breech was delivered with the classical manoeuvre in 10 cases and by the Bracht manoeuvre in 5 cases. Our results show, despite is small case number, that with favourable pelvic scores and absence of cephalopelvic disproportion and fetal distress PgE2 induction can be done locally even for breech presentation. PMID- 9859523 TI - [The mode of delivery in pregnant diabetics]. AB - In our study we have 313 cases--100 cases as a control group and 213 cases as a diabetic group. We studied in these cases the mode of delivery either spontaneous labour, labour induction, cesarean section, forceps and vacuum delivery. PMID- 9859524 TI - [The choice of the uterine-contraction agent in managing the placental period of labor based on our data]. AB - Every obstetrician is acquainted with the existing risk during the placental stage. The danger of a greater blood loss is always present. The use of uterotonic medicines reduces the post-delivery bleeding. The authors have carried out observations over 422 delivery cases. In 153 of them ergotamine has been applied in the placental stage, in 121 oxytocine, and in 148 delivery cases the placental stage has passed without uterotonic drugs. The authors have concluded that the application of contracting medicines reduces the blood loss, but their usage has to be selective and after delivery of the fore shoulder of the foetus. PMID- 9859525 TI - [Multiple pregnancy and microelements: zinc (preliminary report)]. AB - The aim af the study was to define the concentration of zinc in sera of women with multiple pregnancies during the last trimester and to compare it with the values in normal pregnancies. The study included 12 women with gemini and 67 with single fetus pregnancies. A statistically significant decrease was found in the mean zinc concentration in twin pregnancies--8.1 +/- 1.72 mcmol/L compared to the mean values in normal pregnancies--10.5 +/- 1.90 mcmol/L (p < 0.001). Various mechanisms, possibly responsible for the stated changes, their relation to some complications of multiple pregnancy and delivery are discussed, as well as their prophylactics. PMID- 9859526 TI - [Current monophasic hormonal contraception]. AB - Data are presented from an investigation of the use of monophasic contraceptive pills chosen by different age groups. Analyses are made of the data obtained from the Bulgarian Association for Family Planning (BAFP) card index for the last two years. Information is given on the number of observed cycles, type of applied contraceptive pill, aims of their application (contraception, treatment of acne, menstrual disturbances etc.), age of patients, as well as any possible adverse effects--change in blood pressure, bodyweight changes, intermenstrual bleeding, hairiness, breast problems, paraclinical changes, etc. The observed cycles show a very small percentage of adverse effects with hormonal contraception, mainly during the first 2 or 3 cycles, with predominance of intermenstrual bleeding. No bodyweight changes or increased hairiness were observed. PMID- 9859527 TI - [The effect of spinal analgesia on the hemodynamic indices in cesarean section]. AB - We studied systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse frequency (P.F) and mean blood pressure (MBP) among 30 pregnant women in 39.5 +/- 0.3 gestational weeks of pregnancy, age 23 +/- 0.9 years, height 162.0 +/- 1.4 cm and weight 75.5 +/- 2.55 kg. The above indices were studied dynamically in the Ist, 2nd, 3rd, 5th, 7th, 9th, 12th and 15th minute from the onset of the analgesia. We founded that SBP an DBP in the 7th min (the time for spinal block) were lowest. PF and MBP also decreased significantly in the 7th minute in comparison to the inmal values. We can conclude that the haemodynamic indices of the pregnant woman decrease wrong initiation of spinal analgesia for c.s., but remain in the region of normal reference values. PMID- 9859528 TI - [Morbidity in twins with a weight up to 1500 g and a gestational age up to 32 g. w]. AB - Seventy-five newborns from multiple pregnancies with very low and extremely low birth weight are studied. 94% of the infants under 1000 g and 66% of those above 1000 g are born by vaginal way. Intrapartal asphyxia develop most often the second twins with birth weight under 1000 g--64.7%. These are the infants with higher morbidity: RDS--56.5%, IVH--100%. The survival rate of the twins of this group is notably lower than that of the infants from singleton pregnancies with equal weight and gestational age: 12.1% under 1000 g and 69% above 1000 g, against 38.3% and 77.9% respectively. PMID- 9859529 TI - [The effect of antenatal corticosteroid prophylaxis in premature newborns with the use of surfactant therapy]. AB - The study is retrospective aiming to elucidate the effect of antenatal corticosteroids (CS) for prevention of RDS in preterm deliveries, followed by surfactant therapy. 28 premature babies from 26 to 31 gest. weeks, receiving Curosurf at birth, were included in the study. They were divided in two groups: group A-17 babies of mothers with CS prophylaxis and group B-11 babies from deliveries without prenatal CS. All babies were followed for incidence and severity of RDS, duration of artificial ventilation and oxygen therapy. Although mean gestational age and body weight are lower in group A, incidence of severe RDS is lower (23.5% versus 36.4% in group B); duration of artificial ventilation with FiO2 > 0.60 and as a whole is smaller (mean 28.9 hrs and 186 hrs v/s 50.3 hrs and 228 hrs resp. in group B; p < 0.05), duration of oxygen therapy is shorter (mean 428 hrs v/s 540 hrs in group B; p < 0.05), smaller is the incidence of pneumothorax, IVH gr. III-IV, inborn infections and death rate although not statistically significant. The conclusion is made that antenatal use of CS improves lung function in premature babies and potentiates the effect of surfactant. PMID- 9859530 TI - [Cytological and histological parallels in atypical colposcopic findings]. AB - The aim of the present study was to analyse the cytological and histological equivalents in 112 patients with atypical colposcopical findings. The results showed the follow distribution of the findings: for cytological examination in 66 of the patients-I gr. PAP, in 32-II gr. PAP, in 6 III-A gr. PAP, in 1-III-B gr. PAP, in 4-IV gr. PAP and in 3-V gr. PAP. The histological study of the obtained target biopsies established: 7 cases with normal epithelium, 82 with different findings of abnormal epithelium with or without nonspecific and specific inflammatory changes; 18 cases with dysplasia from I to III degrees, 2 cases with CIS and 3 with invasive cancer. The degree of correlation of the different findings, as well as the possible causes for hypo and hyperdiagnostic assessments are discussed. PMID- 9859531 TI - [The ovarian hyperstimulation syndrome in pregnancy after IVF-ET]. AB - The authors present 3 cases of OHSS in 85 patients (3.5%), treated with long stimulation protocol. They propose an original grading of the severity of the syndrome as well as prophylactic and therapeutic measures. PMID- 9859532 TI - [Pelvic actinomycosis]. AB - Actinomycosis is an uncommon inflammatory disease of the female genital tract predominantly affecting the upper sections. Its appearance is in close connection with expended use of different IUDs. Its histological demonstration needs the detection of the specific sulphur granules. Because they often are rare it is recommended working of 8-12 paraffin blocks from suspected cases. The exact diagnosis is of high significance for preventing possible recivides after surgical intervention. PMID- 9859533 TI - [A comparative study of the efficacy of antibiotic prophylaxis with cephalosporins in operative gynecology]. AB - The article presents the results of a comparative study on the efficacy of perioperative prophylaxis with cefalosporins in abdominal gynaecological operations. A total of 155 women meeting certain inclusion criteria were operated on and were randomly assigned to one of the following regimens: Cefoxitin 3 x 1 g -14 subjects, Cefotetan 2 x 1 g--43 subjects, Cefalotin 3 x 2 g for 24 hours--27 subjects, Cefalotin 3 x 2 g for 72 hours--41 subjects and a control group of 30 subjects without prophylaxis. The course of the postoperative period, the hospital stay and the average costs in each group were assessed and compared. None of patients receiving any prophylaxis developed infectious complications, while such complications occurred in 10% of the control without prophylaxis. The authors conclude that antibiotic prophylaxis effectively prevent the infectious complications and shortens the postoperative hospital stay. Despite its narrower anti-bacterial spectrum Cefalotin is not inferior in its efficacy to the second and the third-generation cefalosporins. While providing equal effect, the 24 hours Cefalotin regimen is from 2.8 to 4.9 times cheaper then the other prophylaxis regimens and is twice cheaper than the no-prophylaxis regimen. The longer 72-hours Cefalotin scheme shows no advantages to the 24-hours one but almost triples the expenses. Additional antibacterial treatment in the postoperative period needs more precise indications since unequivocal reasons for its administration are missing in a number of cases. PMID- 9859534 TI - [The diagnostic value of dilatation and curettage in endometrial carcinoma]. AB - The diagnostic accuracy of endometrial curettage is studied in 75 patients treated for endometrial cancer with respect to: 1. Detection of endometrial cancer; 2. Histologic type and grade of the cancer; 3. Spread to the cervical canal. It is found that: 1. The diagnosis of cancer by D&C is correct in 84% of cases; 2. The diagnosis of adenosquamous cancer is made by D&C in 25%; 3. Evidence of spread to the cervical canal by D&C is found in 24 cases and confirm ed by surgery in 7 (29.2%) cases, giving a false positive rate of 70.8%. Negative finding in py with target biopsy is advisable in cases of: 1. Positive cervical finding in the D&C specimen; 2. Negative finding in D&C specimen amd persisting symptoms. PMID- 9859535 TI - [Tumors in childhood and adolescence]. AB - The trial present the results for the incidence and histology of tumours in patients below 18 years, examined at the State University Hospital Maichin Dom in Sofia for two consequent periods of 4 years each. An increase is registered in the incidence of neoplastic disease from 1.85% to 3.42%. There is no significant difference in hystological characteristics of the tumours when comparing I and II period: 85.7% versus 77.4% benign tumours (mucous cysts, dermoid ovarian cysts and polyps) and 14.3% versus 22.6% malignant tumours (ill differentiated teratomas, cystadenocarcinomas, dysgerminomas, ovarian embryocarcinomas and rhabdomyosarcomas). Accurate diagnostics requires detailed gynaecological examination, including a rectal one, vaginal probing, vaginal scopia, ultrasound, laparoscopy. Early discovery followed by surgery provides a possibility for adequate and definite treatment. PMID- 9859537 TI - [The potentials for using cocultivation systems in assisted reproductive technologies]. PMID- 9859536 TI - [Attempts at suicide among girls]. AB - Suicides represent one of the main medico-social problems in Bulgaria. Every day three lives are lost because of suicide in our country. The authors present information on the attempted suicide among young girls from an epidemiological study, provided in 1995. It was found out that the leading motives for suicidal behaviour were the conflicts in the family between the girls and their parents. These conflicts induce fear from punishment, getting sometimes to mishandling. Another part of the suicide attempts were directed to draw their parent's attention, even to bring together separated parents. The frequent cases of premature sexual connections and of sexual misure are discussed. Attention is turned to the group suicide attempts among girl-friends and sister. The authors draw inferences on the necessity of directing the primary prevention among the young girls toward the achievement of mental stability, self-recognition and building up social competence. PMID- 9859538 TI - [The potentials for optimizing cervical screening]. PMID- 9859539 TI - [Updated aspects of hirsutism]. PMID- 9859540 TI - [Disseminated pelvic endometriosis with hyperplastic and metaplastic changes. A case report]. PMID- 9859541 TI - [The testicular feminization syndrome (Morris' syndrome)]. AB - The syndrome of testicular feminization is characterized by female phenotype in the presence of male gonads and has a population frequency of 1:20,000 to 1:60,000. The authors report of 2 cases with this syndrome. Its rarity, diagnostic evaluation and follow-up by the gynecologist are of interest. PMID- 9859543 TI - [Misoprostol--a new prostaglandin in obstetrical practice]. PMID- 9859542 TI - [The treatment of vaginal infections with Macmiror and Macmiror Complex]. AB - The aim of the present study was to examine the therapeutic possibilities of the wide-spectrum medicament Macmiror & Macmiror Complex for the treatment of the vaginal infections. MATERIALS AND METHODS: The study included 52 nonpregnant women among 20 u 54 years (middle age 30.5) with different by kind and intensity colpitis complains. The following microbiological characteristic was established: in 26 cases Candida spp., in 19 Gardnerella vaginalis, in 1 Trichomonas vaginalis and in the rest 6 mixed infection. The treatment of the patients was done in combined scheme: peroral and vaginal administration simultaneously with local treatment of the partner. The control examination was performed bistagely: on 7 10 day and on 30-40 day. RESULTS: The good clinical and microbiological influence of the treated patients was established for the first control examination the effect was found in 89.5% u 84.2% go of the cases with bacterial vaginosis and in 84.6% u 69.2% in those with mycotic colpitis. Relatively high values were found also at the second control, respectively 83.3% and 72.2% for the amine colpitis and 69.5% and 52.2% for vaginal candidosis. The similar favorable influence received also in the rest of the patients. CONCLUSIONS: Our results do show good possibilities of the nifuratel and nystatin combination (Macmiror & Macmiror complex) to influence vaginitis with different etiology, which give us founding to consider, that the drug is suitable for the mixed forms of vaginal infection. PMID- 9859544 TI - [Developmental dysplasia of the hip]. PMID- 9859545 TI - [Pediatric diseases associated with passive smoking]. PMID- 9859546 TI - [Infant mortality and sudden infant death in Navarra between 1985 and 1996]. AB - OBJECTIVE: The objective of this study was to obtain an accurate register of the deaths of children less than one year of age in order to discover the real infant mortality (IM) rate in Navara and its trend during the period between 1985 and 1996, a well as to investigate the deaths by sudden infant death (SID) in the same period and their relationship to the prone position during sleep. PATIENTS AND METHODS: Infant mortality was determined by detailed examination of all certified deaths. Research into possible nonregistered deaths was also carried out. The sources of information included the archives and registers of the pediatric and obstetric wards and pathology departments of all hospitals. Deaths codified as 798.0 (CIE9 infinity R) were included in the study as death by SID. To determine sleeping posture. In the second semester of 1996, a survey was carried out on a sample of 285 children of under 6 months of age. The survey's model is the same as that published previously. RESULTS: In the period of 1985 to 1992, IM was 12.33 per thousand live births. The SID rate was 1.30/1,000 live births. The prevalence of the prone position is unknown. In the period between 1993 and 1996, IM decreased to 5.84/1,000 live births and this decrease was proportional in both the neonatal and postneonatal components. The rate of SIM decreased to 0.44/1,000 live births. At the beginning of 1993, 86% of the children slept in the prone position, in 1994, 38% and in 1996, only 23%. CONCLUSIONS: Trustworthy data in Navarra shows a decrease in IM of more than 50% from 1993 on. The SID rate also decreased by 65%, coinciding with a massive drop in the use of the prone position during sleep. PMID- 9859547 TI - [Group education for asthmatic children: our experience]. AB - OBJECTIVE: Asthma is a chronic disease of great prevalence among children and with complex treatment. Health education is considered a very important therapeutic tool, but it has been poorly assessed in our environment. The main objectives of this work were to assess the utility of the education in groups of asthmatic children and parents in order to improve the knowledge about the disease, to raise the profile of the self-management concept and to improve the clinical evolution. PATIENTS AND METHODS: Sixty-one asthmatic children and their parents were distributed into groups according to the age of the patients (less than 2 years, between 2 and 5 years and more than 5 years) and each group attended 6 theoretical and practical sessions on asthma. Theoretical knowledge about the disease and its management was evaluated by using questionnaires before and after the sessions. The clinical evolution of the children was assessed for a period of two years, from the year before the training started until 1 year after it ended. RESULTS: Theoretical knowledge about the disease clearly improved both in children (p = 0.2) and their tutors (p < 0.00001). The yearly number of crises and hospitalizations were not reduced by the training; however, the number of visits to the outpatient clinic significantly decreased (p < 0.001). Patients started using the flow-meter (p < 0.001) and started proper treatment (p < 0.00001) more frequently after attending the group session. CONCLUSIONS: Group health education for asthmatic children improves both the knowledge and the self management of the disease by the patients and their parents, but does not modify the clinical evolution. PMID- 9859548 TI - [Adverse drug reactions in area I in Asturias]. AB - OBJECTIVE: The objective of this study was to know the incidence of adverse drug reactions in a group of children sent to our office by their pediatrician under the suspicion of having suffered a drug reaction. PATIENTS AND METHODS: Ninety eight children were retrospectively studied. Family background, clinical picture, drugs involved, time elapsing between the onset of symptoms and the date of the study, association to other allergic conditions and diagnostic methods used "in vivo" and "in vitro" were evaluated. RESULTS: Mean age of the patients was 4 years with 53.5% being female and 46.5% male. Positive drug allergy in the family background was found in 22%. The clinical picture showed urticaria and angioedema in 36%, morbiliform eruption in 31%, urticaria in 20%, only angioedema in 9% and other symptoms in 4%. Drugs involved included betalactams (78%), macrolides (17.5%) and others (less than 9%). The mean time elapsed between the onset of symptoms and the date of study was 2 years. There was association to other allergic conditions in 18% (50% in the cases in which the drug allergy was confirmed). RAST to penicillin G and V, amoxicillin and ampicillin was done in 28 cases (betalactams), being negative in all of them. Skin tests were carried out in 21 cases (betalactams), being positive in 19% (66% among the confirmed cases). In all but two cases a controlled provocation test was carried out, being positive only in 8. CONCLUSIONS: 1) The presence of adverse drug reactions is confirmed in only 9% of the cases. 2) Confirmation of the clinical history and the provocation test as the most reliable diagnostic methods currently available. 3) Acceptable sensitivity of the skin tests in the case of betalactams. 4) Null sensitivity of the RAST in the case of betalactams. PMID- 9859549 TI - [Essential bone cyst of the calcaneus in children. Report of three cases]. AB - OBJECTIVE: The purpose of this study was to analyze the properties of essential bone cyst of the calcaneus in childhood, as well as its evolution, and differences with regard to adults. PATIENTS AND METHODS: The cases of children with a diagnosis of essential bone cyst of calcaneus in last sixteen years in the pediatric orthopedic unit of San Carlos Clinic have been reviewed. We evaluated the information regarding the clinical history and the radiological and histological date as well as the therapy result with esteroid intracavity therapy and curettage and bone grafting. The radiologic score was done following Sodergard criteria and the posttreatment follow-up with Neer's score. RESULTS: The mean age of the patients was 10.3 years. Two of the patients had recurrent heel pain for more than six months. Treatment consisted in intracavitary esteroid in two cases and curettage and bone grafting in all three cases, in two of the cases because of failure of esteroid therapy and in the other because of an atypical X-ray image. In all cases the histological stady identified the existence of a characteristic membrane conective tissue. The postsurgery follow up a period of half four years has been satisfactory. CONCLUSIONS: The essential bone cyst of the calcaneous is exceptional localization in children, frequently symptomatic and resistant to the esteroid intracavitaria contrary to what happens with the essential bone cyst of other localizations and with the essential bone cyst of the calcaneus in the adults. The recurrent heel pain and atipical X-ray image are surgery indications. PMID- 9859550 TI - [The impact of psychological preparation for pediatric surgery on postoperative recovery]. AB - OBJECTIVE: Fear of surgery affects recovery in the emotional and behavioral state that leads the child to call the nurse more often or to ingest more sedatives after the operation. The present work evaluated the effects of psychological preparation for surgery during the pre-surgical period on the post-surgical recovery during the hospital stay in order to evaluate the effects that this preparation had on the recovery of the child. PATIENTS AND METHODS: The sample was composed of 60 pediatric patients of both sexes between 7 and 14 years of age admitted for minor scheduled surgery. Fear and five indicators of recovery were evaluated during the post-surgical period (call to the nurse, ingestion of sedatives and liquids, miction and sleep). The children were assigned randomly to one of the following four groups: filmed modeling, coping skills, filmed modeling plus coping skills, and control. RESULTS: The results indicated that the prepared children showed less fear, called the nurses less often, were administered fewer doses of sedatives and slept better after the operation. Analysis of the size of the effect showed that the programs that included coping skills training obtained the most benefits in the post-surgical recovery. PMID- 9859551 TI - [Accidental contact with syringes used by intravenous drug users (IDU): a decade of study]. AB - OBJECTIVE: The increase in drug addiction, AIDS, hepatitis C and B transmission is a problem the society faces with growing frequency, mainly due to syringes and other material used by seropositive patients being left in public place, with children being one of the groups being at greatest risk. We present our experience in accidents with syringes used by intravenous drug users (IDU) in patients less than 16 years of age during the last 11 years. PATIENTS AND METHODS: All patients attending our emergency department with diagnosis of contact with syringes used by IDU were prospectively studied. All patients were treated according to the follow-up protocol of accidental contact with syringes used by IDU. RESULTS: We studied 183 patients between 0 and 16 years of age, with a male gender predominance, the average age at presentation was 6.7 years. The majority of cases occurred during the months of April through August. One hundred twenty-eight cases completed the control after 1 year. At the one year control after the accident, 99% of the patients were anti-HBs antibody positive. Other markers positive for HBV were found in 15% of the cases. Markers for HCV were found in only 9 of 84 patients studied. Only one patient that did not receive either the gamma globulin nor the vaccine developed hepatitis B infection. No patient showed HIV conversion during the time of the study. CONCLUSIONS: The risk of HIV and HCV after accidental puncture with syringes used by IDU is nearly none. Hepatitis B virus infection is the major risk. HBV transmission in the workplace can be largely eliminated due to the increased use of hepatitis B vaccine. PMID- 9859552 TI - [WAGR syndrome: a case report]. AB - OBJECTIVE: WAGR syndrome is a rare syndrome which involves microdeletions of the short arm of chromosome 11 at band 11p13. The clinical features are Wilms' tumor, amiridia, genitourinary abnormalities and mental retardation. There are very few reported cases. We report a new case of WAGR syndrome and review the literature. PATIENTS AND METHODS: Chromosome preparations were obtained from lymphocyte cultures of peripheral blood. For chromosome analysis GTG banding and fluorescent "in situ" hybridization (FISH) were used. RESULTS: Chromosomal analysis revealed deletion of p12-p13 bands. Our patient had bilateral aniridia, Wilms' tumor and cryptorquidia. CONCLUSIONS: The karyotype was 46, XY, del (11)(p12-p13). The p13 band deletion was the cause of the WAGR syndrome. PMID- 9859553 TI - [Mean fetal growth curve adjustment]. AB - OBJECTIVES: To present the optimum equation of the anthropometric fetal growth curve evaluated by height, weight and head circumference according to sex and gestational age. To find the inflection point of the mentioned curves to get to know the precise moment in which growth velocity is the fastest. MATERIAL AND METHOD: We have used the anthropometry of 25,200 newborns (ages ranging from 25 to 43 weeks). All of them were born in the Neonatology Service of the San Carlos Hospital, Madrid. The statistical method used was to check wether fetal growth fitted a logistic curve. The accuracy of the contrast is based on non-linear regression technics. RESULTS: We present the tables for newborns (boys and girls) showing mean growth and weight, height and head circumference upper and lower limits, from the 25th to the 43rd gestational week. CONCLUSIONS: Fetal anthropometric growth fits perfectly a logistic curve. Mean growth velocity reduces sooner in girls than in boys. Weight growth velocity reduces later than height and head circumference. PMID- 9859554 TI - [Neuro-tuberculosis: solitary supratentorial tuberculoma and meningitis in a 5 year-old child]. PMID- 9859555 TI - [Relationship between a case of severe hearing loss and use of gentamycin in the pregnant mother]. PMID- 9859556 TI - [Chronic hiccups in childhood: usefulness of baclofen]. PMID- 9859557 TI - [Epstein-Barr virus, hemophagocytic syndrome and angiocentric lymphoma: a rare and fatal association]. PMID- 9859558 TI - [Smith-Magenis syndrome: a case report]. PMID- 9859559 TI - [Mitochondrial myopathy with a clinical onset simulating Guillain-Barre syndrome]. PMID- 9859560 TI - [Post-chickenpox purpura fulminans with transient reduction of C and S proteins]. PMID- 9859561 TI - [Chronic lumbar pain in an eleven year-old child]. PMID- 9859562 TI - [Eating disorders and the parliament]. PMID- 9859563 TI - [Eating disorders in adolescents: a disease? an epidemic?]. PMID- 9859564 TI - [Prevalence of eating disorders in Spain]. PMID- 9859565 TI - [Facial asymmetry with crying]. PMID- 9859566 TI - [Acute benign viral myositis: neither etiologic confirmation nor complementary proofs]. PMID- 9859567 TI - [Two aspects of pediatrics in Spain. Half a century of existence of the Spanish Pediatric Association (1949-1999) and medical care to children in the Social Security system]. PMID- 9859568 TI - [Acute myeloid leukemia: towards a cure?]. PMID- 9859569 TI - [Mycobacterium tuberculosis drug resistance in patients with HIV and pulmonary tuberculosis infections in Rome: 1987-1996]. AB - A retrospective chart review was performed on 118 HIV infected patients with pulmonary tuberculosis hospitalized between 1987 and 1996 in a tertiary care center for Infectious Diseases in Rome. The aims of this study were: a) to evaluate global prevalence of and risk factors for drug-resistant Mycobacterium tuberculosis and multidrug resistant tuberculosis; b) to assess trends in prevalence of drug-resistant tuberculosis over the 10-year study period. Prevalence of drug resistance of first Mycobacterium tuberculosis isolates was tested on Lowenstein-Jensen medium with the proportional method. Of the 118 patients studied, 83 had never been treated for tuberculosis and 35 had already been treated for at least 1 month. The overall prevalence of resistance to one or more drugs was 25% (17% in never treated patients vs 46% in already treated patients; p = 0.002). Five percent of isolates were resistant to both isoniazid and rifampin (1% in never treated patients vs 14% in already treated patients; p = 0.008). Resistance rates to individual drugs were: isoniazid 14%, rifampin 8%, ethambutol 0%, streptomycin 13%. During the study period no significant variations in prevalence of drug-resistant tuberculosis were found. In our area, empiric therapy should include 4 drugs: as well as isoniazid, rifampin and pyrazinamide, we recommend ethambutol. Surveillance of drug-resistant tuberculosis is needed. Directly observed therapy should be considered for HIV patients in order to prevent increases in drug resistance, relapses, and treatment failures. PMID- 9859570 TI - Long-term results in non randomized patients with acute myelogenous leukemia: a single institution experience. AB - Sixty-three non randomized adults with acute myelogenous leukemia were treated with an idarubicin-based protocol. The patients achieving complete remission received autologous bone marrow transplantation or (if > 50 years or refusing autologous bone marrow transplantation) high-dose Ara-C, as late intensification. Fifty-two patients (82.5%) achieved complete remission, 45 after one induction course and 16 of them underwent autologous bone marrow transplantation a median of 11 months later. As of December 1997 (median follow-up 112 months, range 50 135 months), 16 patients were still in complete remission (10 after autologous bone marrow transplantation, 6 after high-dose Ara-C) and 29 had relapsed (median time to relapse 14 months, range 2-75 months). Four patients died in complete remission. The median disease-free survival was 25 months; the 50-months and 10 year disease-free survival were 41% and 35% respectively. No significant differences were observed between the autologous bone marrow transplantation and high-dose Ara-C treated patients whose complete remission had lasted more than 11 months. The median disease-free survival in the autografted patients had not been reached after 120 months (the 50-month and 10-year disease-free survival chances were both 67%). Age was the only predictive variable for leukemic relapse. These long-term results confirm the antileukemic efficacy of an idarubicin-containing protocol, which led to high complete remission rates and favorably influenced disease-free survival. Furthermore, the efficacy of late intensification treatment with either autologous bone marrow transplantation or high-dose Ara-C is underscored. The disease-free survival chances after autologous bone marrow transplantation are comparable with those published for allogeneic bone marrow transplantation; however, disease-free survival of the patients receiving a high dose Ara-C intensification regimen is not significantly worse than that seen after autologous bone marrow transplantation. PMID- 9859571 TI - [Recurrent aphthous stomatitis: which role for viruses, food, and dental materials?]. AB - Recurrent aphthous stomatitis is a frequently occurring disorder which may be a clinical feature of systemic disease. For many other patients, it is a tedious problem often having no known cause. The aim of this study was to verify if immune responses to common foods and/or viruses are involved in the etiopathogenesis of recurrent aphthous stomatitis. Sixteen patients with this disorder were studied by measurement of immunoglobulin classes (IgG, IgA, IgM), blood lymphocyte subpopulations, blood circulating immune complexes, and complement fractions (C3 and C4). Intradermal skin tests for common food and inhalant allergens were performed in all cases. In 5 patients with positive skin tests, serum specific IgE were tested for the same allergens by radioallergosorbent test. Skin patch tests for dental material were performed in all cases. Oral mucosal biopsies and/or cytology samples were taken in 10 cases for histopathological evaluation and in situ hybridization for Papillomavirus, Cytomegalovirus, Herpes simplex virus I and II, Epstein Barr virus. In 13 patients, lymphocyte subpopulations were altered, with a reduced CD4/CD8 ratio. No other alterations of serum immunological parameters were observed. Skin patch tests for dental material were negative in all cases, while skin tests for food allergens were positive in 5 cases (not confirmed by radioallergosorbent test or food challenge tests). Virus antigen and DNA were not found in mucosal specimens, although one patient was positive for Epstein Barr virus DNA by in situ hybridization. An alteration of the CD4/CD8 ratio was demonstrated in most of the patients with recurrent aphthous stomatitis, although immune responses to food and/or dental material and/or common viruses did not seem to be involved in the etiopathogenesis of this disorder. PMID- 9859572 TI - [Zinc deficiency in liver cirrhosis: a curiosity or a problem?]. AB - This article reviews the literature on the role of zinc in liver cirrhosis and dietary zinc supplementation for cirrhotic patients with hepatic encephalopathy. Zinc is a trace metal found in many proteins and enzymes having catalytical, cocatalytical and structural functions. Zinc deficiency has been demonstrated in cirrhotic patients, and dietary supplementation of this metal could benefit the urea cycle, glucose and protein metabolism, and central nervous system neurotransmission in these subjects. After some brief considerations on normal zinc metabolism, the role zinc plays in the central nervous system, and its conduct in patients with hepatic encephalopathy, possible links between zinc deficiency and the pathophysiology of central nervous system dysfunction in cirrhotic patients are discussed. The article concludes with a critical review of favorable and unfavorable evidence for zinc supplementation in patients with hepatic encephalopathy. PMID- 9859573 TI - [Clinical use of glycoprotein IIb/IIIa inhibitors]. AB - Glycoprotein IIb/IIIa inhibitors are a new class of very powerful antiplatelet agents that act by inhibiting fibrinogen interplatelet bridges. Abciximab, tirofiban, lamifiban, and integrelin, all intravenous formulations, have been evaluated in phase II and phase III clinical trials. Although there seem to be relevant differences among drugs, their efficacy in treating acute ischemic syndromes and preventing acute complications of coronary angioplasty has been demonstrated. The major side effect has been an increase in bleeding complications. Future studies will be aimed at optimizing results and improving safety through more accurate assessment of optimal dosage and duration of infusion. PMID- 9859574 TI - Congenital conduction defects in children born to asymptomatic mothers with anti SSA/SSB antibodies: report of two cases. AB - It is very difficult to identify pregnant asymptomatic mothers carrying anti SSA/SSB antibodies. We report two cases of neonatal lupus erythematosus, born to asymptomatic mothers with anti-SSA/Ro antibody, who developed isolated complete congenital cardiac heart block and transient second degree conduction defect associated with cardiac abnormalities, respectively. The first died suddenly of acute myocarditis at the age of 20 months, while the second underwent surgery at the age of 10 years for a ventricular septal defect, after two episodes of second degree atrioventricular block in infancy. We believe that in both cases the diagnosis could have been made in utero after correct heart beat analysis. We propose careful monitoring of fetal heart beat in all pregnant mothers. The occurrence of heart beat modification should prompt clinicians to test the mother for antibody positivity. This approach may permit early diagnosis and in utero treatment in order to spare the child from cardiac conduction defects. We provide the evidence of these cases and propose a flow chart for all physicians dealing with pregnancy. PMID- 9859575 TI - The "Harlequin Sign". Case description and review of the literature. AB - Asymmetrical facial sweating and flushing has been named the "Harlequin Sign". This is a rare feature, as evident from only 12 cases described up to date. The "Harlequin Sign" represents a local autonomic dysfunction due to a cervical sympathetic deficit located at the pre or postganglionic level on the non flushing side. We observed slow onset and progression of the "Harlequin Sign" in a 19-year-old man, with preexisting slight miosis on the non-flushing side. The differential diagnosis included other forms of dysautonomia and a secondary origin of this partial Horner's syndrome. Both pupils normally reacted to light, convergence, and pilocarpine eye-drop instillation, but the affected side showed supersensitivity to phenylephrine. Deep tendon reflexes were normal, thus excluding Adie's syndrome. The absence of cholinergic supersensitivity in the iris muscles indicated normal function of the ciliary ganglion and excluded the ocular parasympathetic deficit, also evident for the coexisting Horner's syndrome. The clinical features could be explained by assuming that the lesion was located at the level of postganglionic sympathetic fibers, probably due to trans-synaptic postganglionic neuronal degeneration at the level of the stellate ganglion, thus determining the onset of the hemifacial symptoms. PMID- 9859576 TI - Pulmonary hyalinizing granuloma. A limited form of Wegener's granulomatosis? AB - Pulmonary hyalinizing granuloma is an uncommon disease that consists of slowly enlarging nodules in the pulmonary parenchyma. It occurs rarely: in fact, fewer than 70 case reports have been published in the past 20 years. It is important however in the differential diagnosis of lung diseases manifesting multiple pulmonary nodules. The etiology and pathogenesis of this disorder are unknown. Evidence suggests that the nodules could be the result of a chronic exaggerated immune response to infectious agents or to any other process in which antigen antibody complexes are involved. More than 50% of the patients reported have evidence of autoimmune phenomena, e.g. positive antinuclear antibodies, a positive rheumatoid factor, or circulating immune complexes. The present report describes, for the first time, a case of pulmonary hyalinizing granuloma in which the patient had antineutrophil cytoplasmic autoantibodies with a granular cytoplasmatic pattern with typical central accentuation of fluorescence intensity and negative nuclei. The presence of antineutrophil cytoplasmic autoantibodies suggests that pulmonary hyalinizing granuloma could be regarded as a localized, non-evolving, form of Wegener's granulomatosis or a purely granulomatous Wegener's granulomatosis. PMID- 9859577 TI - Lactobacillus casei pneumonia and sepsis in a patient with AIDS. Case report and review of the literature. AB - Lactobacilli are ubiquitous gram-positive anaerobic rods present in the normal bacterial flora of the mouth, vagina and gastrointestinal tract. Although they are usually non pathogenic, serious infections have occasionally been described in transplant recipients or severely ill patients. Only 4 cases have been reported involving AIDS: one had predisposing conditions other than AIDS, and none of the others had pure growth of lactobacilli. We report a case of community acquired Lactobacillus casei pneumonia in a CD4 lymphocyte-depleted AIDS patient. Lactobacillus was isolated in pure growth in repeated blood cultures in an outpatient with no preexisting lung diseases and no known risk factors for Lactobacillus infections (dental procedures, complicated deliveries, gastrointestinal diseases, cardiac prosthetic valves) or consumption of unusual dairy products. Although uncommon, lactobacilli are possible pathogens in HIV infected patients with very low CD4 counts, and their isolation in clinical specimens must not be neglected. Susceptibility tests are essential because of the variable antibiotic-resistance patterns of these bacteria. PMID- 9859578 TI - [Natural history of cancer of the kidney: iconographic follow-up]. AB - OBJECTIVE: To describe the natural history of renal cancer based on the images obtained in a case with more than 11 years' follow-up, from the initial stages of the disease to its outcome. METHODS/RESULTS: The history of a female patient in whom a renal mass had been detected at age 25 years is described. The patient died 11 years later due to metastasis. The images obtained throughout follow-up clearly illustrate the natural course of the disease, which could not be changed because the patient had refused surgery. CONCLUSIONS: The observation of a small renal mass in young patients is an indication for early surgery. The natural course of this disease is predictably fatal, its benign behaviour being highly questionable. Because pregnancy does not appear to accelerate the course of the disease, patient assessment can be completed and treatment can be instituted after delivery. PMID- 9859579 TI - [Does some relation exist in the size and histological composition of the prostate?]. AB - OBJECTIVE: To determine if the histological composition of the prostate-the proportion of glands and stroma-depends on its size. METHODS: Prostate volume was determined by abdominal US; transurethral biopsy of the prostate was performed in 22 patients (mean age 64.8 years) with BPH or bladder tumor. Morphometric analysis of a fragment of the prostate was performed using light and electron microscopy. The stroma-epithelial ratio (SER) and the density of prostatic vascularization (DPV), and the nuclear and cellular areas of 5 epithelial cells in each case, were determined. The ratio of the sum of granular to cellular areas (secreto-cellular ratio), and the nuclear-cytoplasmic ratio (NCR) were determined. The pearson coefficient and linear regression equation that best correlated the prostate volume and the rest of the variables. RESULTS: The mean SER was 4:1 and the DPV was 15.18 vessels/mm2. The cellular and nuclear areas were 79.6 and 21.2 mu2, respectively. The secreto-cellular ratio was 0.083 and the NCR was 0.28. The prostate volume showed no mathematical relationship with SER (r = 0.016, p = 0.9) or DPV (r = 0.026, p = 0.9). Only a random association between the US prostate volume and the cellular and nuclear areas, and the secreto-cellular and NCR could be observed. CONCLUSION: The histological composition of the prostate cannot be determined by the US prostate volume. PMID- 9859580 TI - [Adrenal pseudocysts. Therapeutic attitude]. AB - OBJECTIVE: Cystic tumors of the adrenal gland are uncommon, but are being increasingly more frequently diagnosed during routine radiological evaluation as "incidentalomas". We discuss the differential diagnosis, therapeutic approach and the existing controversies concerning the management of this tumor type. METHODS: Two additional cases of adrenal pseudocyst in two women aged 47 and 38 years are presented. In one case the tumor was discovered incidentally, whereas the other case presented with acute pain arising from intracystic hemorrhage. RESULTS: The fist patient had a cystic tumor of 8 cm with some inner walls. Fine needle aspiration biopsy revealed a benign cystic lesion of the right adrenal gland. At laparotomy, an 8.5 x 4.5 cm multiloculated cystic lesion was excised. The second patient presented with abdominal pain due to intracystic hemorrhage. A Doppler US did not disclose any vessel inside the lesion. We performed a lumbotomy and excised a 7.5 x 6 cm cystic tumor located in the right adrenal gland. Both lesions were diagnosed as adrenal pseudocyst; the second case was a hemorrhagic one. CONCLUSIONS: The therapeutic approach in adrenal cystic tumors can be based upon the radiological and cytological findings since malignant cystic tumors are uncommon. A clear liquid and a negative cytology practically discard malignant tumors. Furthermore, cystic adenocarcinomas are usually large and the cystic liquid is cloudy with abundant cellularity. Surgical treatment is justified in the symptomatic, big or complex tumors (mixed, non-homogeneous). PMID- 9859581 TI - [Should hernioplasty be combined with surgery of the prostate?]. AB - OBJECTIVE: In some patients consulting for a prostatic condition, inguinal hernia might be incidentally discovered during evaluation. The results achieved by simultaneous prostatectomy and hernia repair are presented. METHODS: 52 patients with hyperplasia of the prostate and inguinal hernia underwent treatment for both conditions during the same surgical procedure. Patient ages ranged from 56-88 years. All patients underwent suprapubic prostatectomy and hernia repair. RESULTS: 35 patients had indirect and 17 direct inguinal hernia; 13 were bilateral and 39 unilateral. The hernia had been present from one month of 22 years (mean 46 months). Symptoms related with prostate enlargement had been present from one month to 22 years (mean 40 months). Herniorrhaphy was performed through the inguinal incision in 13 cases, prostatectomy was performed through a midline incision in 50 cases, and the Pfannenstiel incision was utilized in two cases. Pathological analysis showed 49 cases had BPH and 3 had cancer of the prostate. Hernia repair was done using the Bassini procedure in 35 cases, 18 cases had a Mac Vay procedure, preperitoneal herniorrhaphy was performed in one case, and closure and ligation of the sac in 11 cases. There were 15 cases of recurrence (28.8%). CONCLUSIONS: In patients with BPH or prostatic cancer and inguinal hernia, prostatectomy and herniorrhaphy can be performed during the same session without increasing patient morbidity and mortality. Urological surgeons should be familiar with this combined procedure. PMID- 9859582 TI - [Sarcomatoid renal carcinoma]. AB - OBJECTIVE: Sarcomatoid carcinoma of the kidney is a unique, uncommon variety of parenchymatous tumors and is considered to have a worse prognosis by stage than the other histological types. Our experience is reviewed and compared with the larger series reported in the literature. METHODS: Of 101 cases of renal carcinoma submitted to surgery at our department from January, 1990 to December, 1996, there were 4 cases of sarcomatoid carcinoma of the kidney. We compared our cases with the larger series (91 cases in total) reported in the literature for incidence, age, distribution according to sex, tumor size, location, form of presentation, stage and survival. RESULTS: Sarcomatoid renal carcinoma accounts for 1% to 6% of parenchymatous tumors of the kidney according to the different series. The mean age at presentation was 48 years (range 27-61) in our series; there was no prevalence according to sex or location; the renal capsule was not compromised in 3 out of the 4 cases; only one case showed regional lymph node involvement and no case showed distant metastasis at the time of diagnosis. Although these tumors are diagnosed in the advanced stages, two of our cases were incidentally discovered in the early stages (one died 13 months thereafter and the other is alive at 42 months). Patients that are alive and disease-free can only be found in the patient group with stage I or II tumor. All patients with tumor stage III and IV have died. This histological type has a worse prognosis. Most of the series report a mean survival of 5 months. CONCLUSIONS: Sarcomatoid renal carcinoma accounts for 1%-6% of all renal carcinomas. It can present at any age and has a very poor prognosis, with a mean survival of 6 months. PMID- 9859583 TI - [Usefulness of BTA Stat test (Bard) in the diagnosis of bladder cancer. Preliminary results and comparison with cytology and cystoscopy]. AB - OBJECTIVE: The Bard BTA test is an immunochromatographic reaction to detect the bladder tumor antigen in patients with bladder cancer. The sensitivity, specificity and predictive values of this test are analyzed and compared with the standard methods of bladder cancer diagnosis (cystoscopy and cytology). METHODS: In 120 patients with symptoms suggestive of bladder cancer or who came for follow up control evaluation for bladder cancer, we performed the BTA test and cytology in a voided urine sample prior to cystoscopy. TUR and randomized biopsies were performed in patients with a suspicion of bladder cancer. RESULTS: 76 patients had bladder cancer and 44 were free of cancer. The sensitivity was 76.36% for the BTA test, 61.84% for cytology and 98.5% for cystoscopy. The specificity was 77.7% for the BTA test, 86.36% for cytology and 86.36% for cystoscopy. The positive and negative predictive values were 85.29% and 65.38%, 88.68% and 56.72%, and 91.5% and 97.4%, respectively, for each of the foregoing tests. The sensitivity by grade was 43.75% for G1, 78.38% for G2, 95.65% for G3 for the BTA test; 18.75%, 59.46% and 95.65% for G1, G2 and G3, respectively, for cytology. The sensitivity by stage was 53.8% for Ta, 76% for T1, and 100% for T2-4 and Tis for the BTA test, whereas the sensitivity for cytology was 23.08%, 62% and 100%, respectively, for the foregoing tumor stages. CONCLUSIONS: The BTA test is simple, fast and useful in bladder cancer diagnosis. Our results show that the BTA test is superior to cytology. PMID- 9859584 TI - [Valsalva minimal leak point pressure: a useful approximation to type III female urinary incontinence]. AB - OBJECTIVE: It has been reported that anatomic female urinary incontinence with complex sphincteric malposition can coexist with intrinsic damage of the sphincter itself. In this study we analyzed the utility of measuring minimum abdominal pressure at standardized bladder capacities that causes urinary incontinence in order to quantify intrinsic sphincteric damage in female urinary incontinence. METHODS: The study comprised 50 women with urinary incontinence aged 36-78 years (mean 59.4), ICS standardized complete urodynamic study was performed. Minimum leak point pressure with Valsalva maneuver in decumbent and standing positions was determined during the filling phase of cystomanometry and it was defined as a measure of the abdominal pressure expressed as total baldder pressure without involuntary detrusor activity and exercised at standardized bladder capacities that originates objective urinary incontinence. Minimum leak point pressure for each bladder capacity was evaluated. Leak point pressures below 60 cm H2O indicate intrinsic sphincteric damage; pressures between 60 and 90 cm H2O indicate intrinsic damage and malposition or urethral hypermobility may coexist, and leak pressures over 90 cm H2O are related to complex sphincteric malposition. RESULTS: 5 women showed severe sphincteric deficiency (type III) and urinary incontinence was demonstrated with 50 ml bladder capacity and 30 cm H2O of abdominal pressure without detrusor activity. Thirty-five women (70%) had type II urinary incontinence. Of these, 10 (28.5%) showed intrinsic sphincteric damage in addition to malpositioning of the sphincteric complex at leak point pressures between 60 and 90 cm H2O. The rest of the women showed Blaivas' type 0 and I urinary incontinence. CONCLUSIONS: Valsalva minimum leak point pressure is a reproducible, reliable, useful and easily measured parameter in diagnosing female stress urinary incontinence. It allows approximation of the abdominal pressure to the level at which urinary leakage is produced during the filling phase of cystomanometry and gives us an idea of the extent of the intrinsic sphincteric damage, if any. Not only is sphincter damage demonstrated in type III urinary incontinence, but that it may also coexist to a varying degree with complex sphincteric malposition. PMID- 9859585 TI - [Functional reinnervation in patients with a diagnosis of lower motor neuron neurogenic bladder: prognostic and therapeutic considerations]. AB - OBJECTIVE: To evaluate bladder and urethral sphincter reinnervation mechanisms during long-term follow-up in patients with lower motor neuron neurogenic bladder following neurological surgical injury. METHODS: A urodynamic study was conducted in 30 patients (21 male and 9 female; mean age 53.4 years) with lower motor neuron neurogenic bladder dysfunction arising from neurological injury sustained during surgery. The protocol included cystometry and periurethral electromyography (EMG) at 3, 6, 9, 12 months and once a year for 7 years, and videocystography at 3, 12 months and once a year for 7 years. Functional parasympathetic (detrusor) reinnervation criteria were cystometric. Functional sympathetic (bladder neck) reinnervation criteria were cystographic. Functional pudendal (periurethral sphincter) reinnervation criteria were electromyographic (increase of polyphasic and long amplitude and/or long time potentials). RESULTS: Detrusor reinnervation was demonstrated in 6 male patients (20%) with an average period of 44.6 months. Pudendal reinnervation was demonstrated in 17 patients (77.2%) with an average period of 17.6 months. EMG potentials were polyphasic in 17 cases and long amplitude/long time potentials in 3 cases. Sympathetic reinnervation was demonstrated in one patient (16.6%) at 60 months. CONCLUSIONS: Functional pudendal reinnervation of the periurethral sphincter was more frequent and was demonstrated earlier than reinnervation in vegetative elements (parasympathetic and sympathetic). Parasympathetic reinnervation had long-term therapeutic implications. Ongoing urodynamic assessment in patients with lower motor neuron neurogenic bladder following abdominoperineal resection or intervertebral disc prolapse surgery is warranted. Sympathetic reinnervation was scanty and was demonstrated later in relation to distal postganglionic fibers. PMID- 9859586 TI - [Urodynamics of urinary incontinence post radical prostatectomy]. AB - OBJECTIVE: To determine the urodynamic characteristics of urinary incontinence after radical retropubic prostatectomy. METHODS: We conducted a clinical and urodynamic study on 25 patients who had undergone radical retropubic prostatectomy due to localized prostate adenocarcinoma. RESULTS: 68% of the patients completely recovered urinary continence. The maximum rate of urinary continence recovery was found between the third and fourth month postprostatectomy. A relationship was demonstrated between preoperative hormone blockade, duration of the surgery and urinary continence. Videocystography demonstrated an incompetent proximal continence mechanism in all radical prostatectomy patients. All incontinent patients presented stress urinary incontinence. Denervation potentials of the periurethral sphincter was demonstrated by electromyography in 80% of incontinent patients. These potentials were not present in the continent patients. Reinnervation potentials were present in 50% of the continent patients and in 20% of the incontinent patients (significant differences). No relationship was found between other urodynamic data and post-radical prostatectomy urinary incontinence. CONCLUSIONS: The surgical difficulty influences the preservation of urinary continence. A high percentage of patients submitted to radical prostatectomy recover urinary continence with time. Urinary incontinence following radical prostatectomy is based on the distal sphincteric mechanism. Patients who remain incontinent four months postoperatively and with electromyographically demonstrated denervation potentials of the periurethral sphincter can be considered to be candidates for treatment of incontinence without waiting any further. PMID- 9859587 TI - [Comparative study of determined biochemical parameters in semen of vasectomized and non-vasectomized subjects]. AB - OBJECTIVE: Biochemical parameters, fundamentally those of prostatic origin, were analyzed and compared to determine the semen composition of vasectomized and non vasectomized subjects. METHODS: 100 ml of semen from each group (vasectomized and non-vasectomized) were obtained. After thawing, the pool was homogenized and 5 one-ml sample from each one were analyzed for the following biochemical parameters: gamma glutamyl transferase (GGT), lactic dehydrogenase (LDH), prostate acid phosphatase (PAP), prostate-specific antigen (PSA) and total protein (TP) in the seminal plasma. Determination of the foregoing biochemical parameters was performed automatically with autoanalyzers. RESULTS: The mean values for the vasectomized and non-vasectomized groups were respectively: a) GGT: 8890 +/- 811 and 5714 +/- 403 IU/L; b) LDH: 7045 +/- 879 and 2465 +/- 339 IU/L; c) PAP: 2,099,000 +/- 330,764 and 1,860,000 +/- 302,138 ng/ml; d) PSA: 953,000 +/- 154,715 and 1,018,000 +/- 119,685 ng/ml; total protein: 39,860 +/- 1094 and 37,900 +/- 5606 micrograms/ml. CONCLUSIONS: Analysis of the different biochemical parameters showed no statistically significant differences between the mean values for PAP, PSA and total protein for the vasectomized and non vasectomized group, but statistically significant differences were found for GGT and LDH (p < 0.001). PMID- 9859588 TI - [Uretero-iliac fistula]. AB - OBJECTIVE: To describe a case of uretero-iliac fistula, an extremely rare condition that is not easily diagnosed. METHODS/RESULTS: Herein we describe a 76 year-old female who had undergone repeated pelvic surgery for adenocarcinoma of the sigmoid. She had a right ureteral fistula that had been managed conservatively by insertion of a ureteral catheter. Diagnosis was made by selective arteriography of the iliac arteries. The patient was submitted to surgery; primary closure of the arterial fistula and nephroureterectomy were performed. CONCLUSIONS: Uretero-iliac fistula should be suspected in patients with a history of repeated pelvic surgery and ureteral catheter placement that present with massive hematuria. PMID- 9859590 TI - [Keratotic pseudoepitheliomatous and micaceous balanitis]. AB - OBJECTIVE: To describe the clinical and histopathological manifestations of a case of pseudoepitheliomatous, keratotic and micaceous balanitis and review the literature on this rare condition. METHODS: A case of pseudoepitheliomatous, keratotic and micaceous balanitis in a 63-year-old man is presented. The lesion appeared a few months after undergoing circumcision due to phimosis and recurrent inflammatory balanitis. RESULTS: The lesion relapsed after topical treatment with 5-FU, cryotherapy and shaving-biopsy plus electrocoagulation. It was finally diagnosed as epidermoid carcinoma and surgically excised. CONCLUSIONS: The clinical and histopathological features, course and relationship between micaceous balanitis and carcinoma of the penis are reviewed. PMID- 9859589 TI - [Hybrid tumor of the penis: is this denomination correct?]. AB - OBJECTIVE: Few cases of verrucous carcinoma of the penis with foci of invasive squamous cell carcinoma have been reported and denominated "hybrid tumors". The accuracy of this term is discussed in this paper. METHODS/RESULTS: A huge penile mass in a patient that had undergone three previous operations for lesions diagnosed as verrucous carcinoma is reported. Partial penectomy was performed. Histological examination showed a very well-differentiated squamous cell carcinoma. PCR (polymerase chain reaction) did not detect any type of human papillomavirus (HPV) in the tumor. CONCLUSIONS: Verrucous carcinoma is a strictly defined lesion with a different biological behaviour from that of squamous carcinoma. Preoperative deep biopsy may miss the squamous cell carcinoma. Definitive diagnosis can only be achieved by histological examination of the surgical specimen. In future, DNA studies could possibly support preoperative diagnosis of this lesion. PMID- 9859591 TI - [Epidermoid cyst of the testicle: presentation of a case]. AB - OBJECTIVE: To describe a case of epidermoid cyst of the testis, a rare lesion that warrants surgical exploration to confirm the diagnosis since its echo patterns are characteristic but not pathognomonic. METHODS/RESULTS: 10 years earlier the patient had noticed an asymptomatic swelling of the right testis which did not cause infertility since he had two children over the last 6 years. Tumor markers AFP, BHCG and LDH fell within the normal ranges. Ultrasound evaluation with a high resolution 7.5 Mhz transducer revealed a 4 x 3.5 x 3.2 cm testicular mass surrounded by atrophic parenchyma. The patient underwent radical inguinal orchidectomy. CONCLUSIONS: Epidermoid cyst is the benign variety of testicular teratoma. Two different forms exist: simple and mixed. Malignant components are present in the latter form. If the simple form is identified before substitution of the testicular parenchyma occurs, testis sparing surgery is justified; otherwise radical orchidectomy with inguinal exploration is indicated. PMID- 9859592 TI - [Risk of tumor seeding after nephroureterectomy combined with endoscopic resection of the ureteral meatus]. AB - OBJECTIVE: To describe a case of tumor implantation at the site of resection in the ureteral orifice, following nephroureterectomy combined with resection of the bladder cuff of. METHODS/RESULTS: A patient with high grade and stage carcinoma of the renal pelvis and no previous bladder tumor presented with a perivesical mass 6 months following nephroureterectomy. She had a high grade transitional cell carcinoma that infiltrated all of the bladder wall, vagina and parametrium in the region of the meatus that had been resected endoscopically. CONCLUSIONS: The present case indicates there may be histological or technique-related factors that facilitate tumor cell implantation in the deep perivesical region after this procedure is performed. It is necessary to identify these factors before this procedure is used widely. PMID- 9859593 TI - [Fracture of the penis: value of echo-Doppler-color]. AB - OBJECTIVE: A case of traumatic rupture of the corpora cavernosa evaluated by color Doppler ultrasonography is presented. METHODS: A 43-year-old male consulted for a large penile hematoma that had presented 16 hours earlier during sexual intercourse. The patient was evaluated by Doppler ultrasound using a 5 Mhz linear probe. The vascular integrity of the penis was demonstrated by color Doppler ultrasonography of the cavernous arteries and penile veins. RESULTS: Disruption at the base of the right corpus cavernosum associated with a small hematoma appeared as adjacent hypoechoic images. The color Doppler ultrasonic evaluation demonstrated the integrity of the arteries and cavernous veins and no pulsation of the hematoma, indicating conservative management. Four months after the trauma, no changes in erectile function or penile deviation have been observed. CONCLUSIONS: Color Doppler ultrasound is a useful diagnostic imaging technique in the assessment and follow-up of penile trauma. PMID- 9859594 TI - Urinary symptoms in women with gynecological disorders: the role of symptom evaluation and home uroflowmetry. AB - OBJECTIVE: The aim of our study was to analyze the effect of gynecological dysfunction on voiding symptoms in women. METHODS: A modified AUA symptom index questionnaire was self-administered and an ambulatory home uroflowmetry was performed, using a specially designed home uroflowmetry apparatus for multiple flow measurements. Urinary symptoms and home uroflowmetry (Home Urodata TM System) were evaluated in 68 women: 34 patients with gynecological dysfunction, and 34 normal controls. A total of 156 urination episodes was recorded, with a mean of 5.4 measurements per patient. RESULTS: Symptom index showed lower values in the group of normal controls than in the group with gynecological dysfunction. Voided volume, peak flow rate and average flow rate were all significantly better in the group of normal controls than in the group with gynecological disorders; the most remarkable changes were observed in patients with genital prolapse and with large uterine fibroma. In both groups the total urine volume was lower between midnight and 8 a.m. (2460 ml), if compared with the total urine volume between 8 a.m. and 4 p.m. (3360 ml) and the total urine volume between 4 p.m. and midnight (3072 ml) (p < 0.05). CONCLUSIONS: Noninvasive home uroflowmetry combines the information of a typical flowchart with uroflow parameters and supplies the physician with multiple consecutive voiding episodes, minimizing the environmental artifacts of the study. In our experience it was found to be useful to evaluate urinary symptoms reported by 18/34 patients (52.9%) with gynecological dysfunction. It can help to select which patients require further urodynamic investigation to improve the diagnostic accuracy and choose the correct treatment. PMID- 9859595 TI - In vitro properties of ciprofloxacin suppositories formulated with glycerogelatin and theobroma oil bases. AB - Ciprofloxacin suppositories formulated with glycerogelatin approximately nd theobroma oil bases were evaluated for in vitro bioavailability in 0.01N sodium hydroxide (pH approximately 10) and in phosphate buffer (pE- approximately 7.8). The release of ciprofloxacin from the two bases under conditions which simulated the conditions in the rectum was assessed using isolated pig rectum. The release of ciprofloxacin was faster from theobroma oil base than from glycerogelatin base using 0.01N sodium hydroxide as the dissolution medium. The cumulative amount of ciprofloxacin released was markedly reduced from each of the two bases in the presence of isolated pig's rectum and in phosphate buffer. PMID- 9859596 TI - Lipidic matrix of albendazole: an alternative for systemic infections. AB - Albendazole is a poorly water soluble drug, with low oral bioavailability, used in pharmacological treatment of a systemic disease as hydatid parasitosis. Lipidic matrices of Gelucires (44/14 and 35/02) were developed. After "in vitro" studies, one formulation was chosen for a single dose study in 8 healthy volunteers, with a cross-over and randomised design, taking a commercially available tablet as reference. Drug absorption was followed by albendazole sulphoxide dosage in urine by high pressure liquid chromatography. Neither albendazole nor albendazole sulphoxide were recovered in urine after tablet administration while 0.18% (+/- 0.06) of dose was recovered after lipidic matrix administration in the first 24 hours. Besides ageing control were performed up to 18 months post-elaboration. Lipidic matrix with Gelucire 44/14 was revealed as a promising attempt for oral pharmaceutical form in albendazole systemic treatment. PMID- 9859597 TI - New 1,2,3-triazole derivatives (ureas, amides, urethanes) with a potential biological interest. AB - This paper reports the preparation of three series of 1,2,3-triazole derivatives bearing an ureido substituent (compounds 5a-h), a carboxamido substituent (compounds 6a-f) or an urethane substituent (compounds 7a-l). Some compounds were submitted to in vitro functional tests on guinea-pig isolated intestinal preparations and/or to in vitro antitumor and antiviral screening. The tested compounds showed no activity on guinea-pig ileum, except compound 7g, provided of contracturant properties; similarly, no anticancer or antiviral significant activity was found. PMID- 9859598 TI - 1,1-Bis-(1,2,4-triazolyl and 1,3,4-thiadiazolyl) ethane and phenylethane derivatives as potential antibacterial agents. AB - A series of compounds containing two identical heterocyclic moieties such as triazole and thiadiazole separated by ethane or phenylethane was synthesized. The structures of the newly prepared compounds were established on the basis of their elemental analyses and spectral data. The target compounds were subjected to antifungal and antibacterial screening. The results of this testing revealed that compounds 23, 26, 34 and 35 showed promising antibacterial activity, while, compound 33 showed remarkable antifungal activity. PMID- 9859599 TI - [September, let's go: it is the time to migrate]. PMID- 9859600 TI - [New systems of cardiac mapping or life to the electrophysiologist may become complicated]. PMID- 9859601 TI - Minimally invasive coronary surgery at the beginning of the new millennium. PMID- 9859603 TI - [Pre-infarction angina and post-thrombolysis recanalization]. PMID- 9859602 TI - Endothelin-1 in heart failure: does it play a role? PMID- 9859604 TI - Brain natriuretic peptide in hypertrophic obstructive cardiomyopathy. PMID- 9859605 TI - Central role of peripheral mechanisms in exercise intolerance in chronic heart failure: the muscle hypothesis. PMID- 9859606 TI - [Reversible postischemic left ventricular dysfunction: treatment by coronary angioplasty]. PMID- 9859607 TI - [Etiopathogenesis of acute myocardial infarction: role of early leukocytosis]. AB - It has recently been suggested that inflammation may play an important role in the pathogenesis of acute ischemic syndromes. It may therefore be important to relate their clinical features with plasma indexes of inflammation. We have studied leukocyte, platelet and fibrinogen blood levels in 57 consecutive patients with acute myocardial infarction admitted to our Intensive Care Unit within 90 min after the onset of chest pain and treated with primary coronary angioplasty. Patients were divided into two groups on the basis of blood leukocyte levels: Group A, 24 patients, 17 males, mean age 54.2 +/- 13.7 years, with high blood leukocytes and Group B, 33 patients, 28 males, mean age 60.9 +/- 10.3 years, with normal blood leukocytes. Group A patients also had higher serum fibrinogen (p = 0.05) and blood platelet levels (p < 0.05). The stenosis observed after guidewire advancement was significant (> 75%) in 33% of the patients with leukocytosis vs 94% of the others (p < 0.01). No difference between the two groups was observed in the success rate of coronary angioplasty and prevalence of stent placement (100 vs 97%, and 43 vs 42% of the patients of Group A and B, respectively). In contrast, a tendency to rethrombosis requiring Rheopro administration was observed in 62% Group A patients vs 21% Group B patients (p < 0.01). In conclusion, the finding of leukocytosis in the acute phase of myocardial infarction suggests that coronary occlusion is mainly caused by a coronary thrombus occurring at the site of a non significant stenosis. In contrast, when blood leukocytes are normal, the underlying coronary stenosis is more often critical and the thrombotic process is less important. The high blood leukocytes, platelet and fibrinogen levels of Group A patients are consistent with a significant role of inflammation in the pathogenesis of the thrombotic process while hemodynamic and local mechanical factors are probably more important in patients with normal blood leukocytes. PMID- 9859608 TI - [Pulmonary hypertension in patients with left ventricular dysfunction studied with contrast-enhanced Doppler echocardiography: relations with diastolic parameters and prognostic implications]. AB - In patients with left ventricular dysfunction, the prognostic value of both pulmonary hypertension and mitral flow patterns has been recognized. However, the effect of the association of different degrees of pulmonary hypertension on prognosis and the corresponding left ventricular diastolic dysfunction is not clear. Accordingly, we considered the impact on survival of a categorization based on the relationship between pulmonary artery pressure and left ventricular diastolic dysfunction, as assessed by mitral and pulmonary venous flow analyses. Transthoracic Doppler echocardiography was carried out in 92 patients with ejection fraction < 45%, pulmonary artery systolic pressure > 25 mmHg and sinus rhythm. Tricuspid regurgitant velocity and Doppler parameters derived from transmitral and pulmonary venous flows were evaluated. In the case of inadequate baseline tracings, weak or poor Doppler signals were enhanced by intravenous injection of a galactose-based contrast agent (Levovist 8 ml suspension at a concentration of 400 mg/ml). To select those whose pulmonary hypertension was either proportional or unproportional to left side filling pressures, patients were divided as follows: Group 1 (n = 69) with low discrepancies and Group 2 (n = 23) with marked discrepancies between Doppler estimates of pulmonary artery systolic pressure and left side filling abnormalities. The patients of each group were also classified according to their mitral flow pattern: abnormal relaxation, pseudonormal and restrictive. Mean pulmonary artery systolic pressure was 49 +/- 16 mmHg in the total population, 43 +/- 11 mmHg in Group 1 and 68 +/- 14 mmHg in Group 2 (p < 0.0001). Several mitral and pulmonary venous flow variables significantly correlated with pulmonary artery systolic pressure in the total population and in the study groups. The best correlations were observed in Group 1 as regards the ratio of reverse-to-forward atrial wave duration (r = 0.83), E wave deceleration rate (r = 0.81), E wave deceleration time (r = -0.81) and the systolic fraction of pulmonary venous flow peak velocities (r = -0.75). In Group 1, the lower heart failure-free survival rate at 10 months was observed in patients with restrictive pattern (68%) as opposed to those with pseudonormal (94%) and abnormal relaxation patterns (97%). The overall heart failure-free survival rate in Group 2 was 86%. In conclusion, the classification according to the relationship between pulmonary hypertension and the alterations of left chamber filling may contribute to the prognostic stratification of patients with left ventricular dysfunction. The patients with pulmonary hypertension proportional to the increase in left chamber filling pressures and restrictive pattern exhibited the worst prognosis. PMID- 9859609 TI - [Natural harmonic echocardiography]. AB - The acquisition of echocardiographic images in harmonic mode (a frequency double than the transmitted, or fundamental) improves imaging quality. We assessed whether harmonic imaging improves the detection of endocardial borders, evaluation of ventricular function and diagnostic confidence in the clinical arena. We have studied in fundamental and harmonic imaging 45 patients (age 20-89 years, mean 53 years) using a multifrequency transthoracic probe transmitting at 1.75 MHz and receiving at 3.5 MHz (Acuson Sequoia). In 34 low echogenic patients we assessed left ventricular function. The remaining 11 patients represented selected cases (i.e. atrial septal aneurysm, aortic dissection, endocarditis and atrial septal defect). The echocardiographic images were recorded on a magneto optical disk and analyzed by two blinded observers. Endocardial definition has been semiquantitatively evaluated assigning a 0-4 score for each of the 16 segments of the left ventricle. A score of 0 was allotted to the non-visualizable segments and a score of 4 to the best detectable segments. Ejection fraction was calculated in each patient from the apical 4-chamber view. We compared endocardial border definition and ejection fraction at rest, in fundamental and harmonic mode, and assessed the interobserver agreement in the calculation of ejection fraction. Harmonic images always showed a better definition and lower noise compared to fundamental. Endocardial border definition was significantly improved in all segments (from 1.3 +/- 1.1 fundamental to 2.9 +/- 1.0 harmonic). Forty-two segments were non detectable in fundamental (score 0) compared to 5 in harmonic. Of these 42 segments, 37 were detectable in harmonic, with a score of 2.0 +/- 1.0. Conversely, none of the 5 segments non detectable in harmonic could be visualized in fundamental. The interobserver agreement in calculating ejection fraction was improved by harmonic imaging compared to fundamental (r = 0.91 and r = 0.67, respectively). In the selected clinical cases the diagnosis was easier and faster by harmonic imaging. The harmonic mode drastically improves echocardiographic imaging, it may be used routinely and reduce the need for more invasive techniques such as transesophageal echocardiography. PMID- 9859610 TI - Hyperplasia of the aorticopulmonary paraganglia: a new insight into the pathogenesis of sudden infant death syndrome? AB - In this study, the Authors search for morphologic alterations in the aorticopulmonary paraganglia (APP) of sudden infant death syndrome (SIDS) victims when compared with age-matched controls. Morphometric studies on serial sections of the APP were performed with an image analyzer in combination with a standard microscope attached to a video camera. APP hyperplasia, characterized by an increase in some parameters such as number, mean lobule diameter and total glomic tissue volume when compared with age-matched controls, was observed in 23.8% of SIDS victims. Similar alterations have been reported in peripheral chemoreceptors of animals and human beings who are chronically hypoxemic. In SIDS, it could reflect an abnormal chemoreceptor function, contributing to an altered respiration control. PMID- 9859611 TI - [Dolichoarteriopathies (kinking, coiling, tortuosity) of carotid arteries and atherosclerotic disease: an ultrasonographic study]. AB - The clinical significance of the dolichoarteriopathies (kinking, coiling and tortuosity) of the extracranial carotid arteries, their prognosis and etiopathogenesis have not yet been clearly defined. The aim of this study was to evaluate, by echo-color-Doppler, atherosclerotic lesions of carotids in subjects with, compared to those without dolichoarteriopathies. A total of 1220 subjects (aged 25-89 years), 316 with and 904 without dolichoarteriopathies, were studied. Patients were divided into 325 subjects without cardiovascular risk factors and 895 subjects with cardiovascular risk factors (352 with arterial hypertension alone, 298 with arterial hypertension associated with other cardiovascular risk factors, 245 with hypercholesterolemia, diabetes, cigarette smoking, alone or associated). Moreover, the study was performed specifically in a subgroup of 352 subjects with arterial hypertension without other cardiovascular risk factors. Myointimal thickness, as a mean of 5 measurements at the level of the common carotid, 2-3 cm from the flow-divider and plaques as a focal thickness > or = 2 mm, classified on the basis of echogenic characteristics (hard, fibrous, mixed, soft, hemorrhagic) and seat (bifurcation, common, internal and external carotid) were evaluated. In 316 patients with dolichoarteriopathies, compared to 904 patients without dolichoarteriopathies there were no differences in myointimal thickening (NS), the prevalence of carotid plaques was lower (p = 0.004) and there was no difference (NS) regarding ultrasonic characteristics (more frequently hard and less frequently fibrous, mixed and soft) and localization of plaques (more frequently at the level of the common and bifurcation and less frequently at the level of the internal and external carotid). In subjects without cardiovascular risk factors, myointimal thickness and carotid plaques did not show any significant differences (NS) in the group with compared to the without dolichoarteriopathies. On the contrary, either in subjects with cardiovascular risk factors, or in the subgroup with hypertension alone, myointimal thickening was equally represented (NS), while carotids with plaques were significantly less frequent (p = 0.002; p = 0.01) in the group with, compared to that without dolichoarteriopathies. In conclusion, dolichoarteriopathies of the carotids do not seem to be a consequence of atherosclerotic lesions but in the presence of cardiovascular risk factors or hypertension alone they may even prevent plaque formation. PMID- 9859612 TI - Constrictive pericarditis presenting as unexplained recurrent pleural effusion: a case report. AB - Constrictive pericarditis may exceptionally present as pleural effusion of unknown origin and this form of presentation may cause diagnostic problems. We report a case of subacute constrictive pericarditis in which there were recurrent pleural effusion with no other signs of the disease and the initial echocardiographic study was nondiagnostic. For this reason the patient was initially considered to have primary pulmonary or pleural disease. On the basis of the subsequent development of signs of systemic congestion and the results of computed tomography, Doppler echocardiography and cardiac catheterization, which were consistent with constriction, it was concluded that the patient had constrictive pericarditis. A complete resolution of pleural effusion and signs of systemic congestion was observed following pericardiectomy. PMID- 9859613 TI - [Lipoprotein plasmapheresis]. PMID- 9859614 TI - [Prevention of opportunistic infections in the protease inhibitor era]. AB - From the middle of 1996 we are living a striking reduction of incidence of opportunistic infections (Ols) associated to human immunodeficiency virus (HIV). The recovery of the immune system, at least partially, is showing up substantial changes of Ols after the introduction of highly active antiretroviral therapy (HAART): relieves, sometime complete resolutions, of Ols that previously did not give any response to the treatment (cryptosporidiosis, microsporidiosis, progressive multifocal leucoencephalopathy and Kaposi's sarcoma), changes of clinical presentations after HAART (CMV retinitis [CMVR] with vitritis and Mycobacterium avium-intracellulare [MAC] lymphadenitis), related to exuberant inflammatory response; and at last, long periods without reactivation of the Ols after prophylaxis suppression (CMVR and Pneumocystis carinii pneumonia [PCN]). All this sep up the necessity of a change in the prophylaxis recommendations after HAART introduction. This change would have been unthinkable two years ago, the point is to answer the following question: when can Ols prophylaxis to be stopped after HAART? The progress in the therapy of HIV and Ols infections have happened that fast that this recommendations will have to be reconsidered continuously. PMID- 9859616 TI - [Diagnosis and treatment of mycobacterial infections in patients with HIV/AIDS]. AB - Mycobacterial infection remains as a frequent complication associated to HIV infection. Although the widespread use of HAART has intensely decreased incidence of disseminated Mycobacterium avium (MAC) infection, it does not seem that it has affected tuberculosis occurrence so intensely. In spite of the intense search of new methods of rapid diagnosis, in the clinical practice the diagnosis of the mycobacterial illnesses continues based on culture, although the appearance of new media has allowed to shorten the time of growth. The combination of isoniazid (INH), rifampin (RIF) and pirazinamide (PZ) (with ethambutol [ETB] when primary resistance to INH is higher than 4%), remains as the elective treatment for tuberculosis in HIV infected patients. Due to the interaction between RIF and some antiretovirals drugs, such as proteasa inhibitors, a change in the usual regimens could be necessary. Combinations without RIF or antiretroviral therapy with drugs not interacting with RIF (nucleosides, ritonavir or nevirapin) have been suggested. The emergence of strains of Mycobacterium tuberculosis resistant to the antituberculosis drugs, the lack of adherence to treatment, and the frequency of adverse events hinders even more the control of the tuberculosis and they demand a narrow follow up of these patients. The treatment of the disseminated infection by MAC has improved in the last years with the generalization of the combinations including macrolides as claritromicin or azitromicin with ETB. The doubt persists about what combination is more effective, although like in other opportunists infections associated with a severe immunodeficiency, using antiretrovirals combinations that enhance the immune system could be a fundamental therapeutic approach. PMID- 9859615 TI - [Advances in the diagnosis and treatment of infections caused by herpesvirus and JC virus]. AB - During the last two years important advances in the diagnosis and treatment of cytomegalovirus (CMV) disease have occurred. Several studies have suggested that biologic markers of CMV viremia (PCR, branched DNA and pp65 antigenemia) might be useful both to stratify the risk of developing CMV disease and to follow the response of CMV retinitis to therapy. It has been shown that patients who are plasma CMV PCR positive have a risk of developing CMV disease three times higher than patients who are plasma CMV PCR negative. In addition, for each log10 increase of the CMV viral load there is a 3-fold higher risk of developing CMV disease. Currently, therapeutic options for induction treatment of CMV retinitis (CMVR) are: i.v. ganciclovir (GCV), i.v. foscarnet, i.v. cidofovir or GCV intraocular implant combined with oral GCV. For maintenance therapy options are: i.v. GCV (3, 5 or 7 days per week), oral GCV (only for peripheral retinitis), i.v. foscarnet (daily), i.v. cidofovir (biweekly) and GCV intraocular implant (replaced every 6-8 months) combined with oral GCV. There is currently enough evidence to allow the diagnosis of progressive multifocal leukoencephalopathy (PML) based on the finding of JC virus DNA in CSF by PCR. There are still no drugs with proven clinical efficacy against JC virus but the possibility that HAART treatments might improve the control of this disease appear promising. PMID- 9859617 TI - [Infections by Candida and cryptococci]. AB - The introduction of highly active antiretroviral therapy (HAART) has reduced dramatically the incidence of mucosal candidiasis and cryptococcosis in AIDS patients. Fluconazole is the drug of choice for candidiasis. The duration of antifungal treatment is based on response, but typically 7 to 14 days are required for oropharyngeal forms and up to 21 days for esophageal disease (200 the first day and 100 mg thereafter). Resistant candidiasis tends to occur in persons with advanced HIV disease and previous fluconazole therapy who have been noted to result in clinical improvement with HAART. HAART must be considered the therapy of choice for refractory candidiasis. The preferred treatment for cryptococcal meningitis includes two weeks induction treatment with amphotericin B (0.7 mg/kg/d IV) with or without flucytosine (25 mg/kg qid) followed by 8 weeks of fluconazole (200-400 mg PO qd). Long-term maintenance therapy with fluconazole (200-400 mg PO qd) is required to prevent relapses. In patients with elevated intracranial pressure who had focal neurologic deficits or mental status changes, serial lumbar punctures should be performed. In refractory cases the immediate placement of CSF drains must be considered. PMID- 9859618 TI - [Pneumocystis carinii pneumonia and HIV infection: diagnosis and treatment]. AB - Pneumocystis carinii pneumonia (PCP) is one of the leading complications among HIV-infected patients. Recent advances in PCP prophylaxis, diagnosis and treatment have caused a decrease in PCP-related morbidity and mortality. Despite these advances, PCP continues to be frequent in patients not known to be HIV infected and in those patients with poor adherence to prophylactic regimens or severe immunosuppression. In typical cases diagnosis may be suspected by the patient's clinical presentation. Clinicians are frequently faced with the differential diagnosis between PCP, bacterial pneumonia, pulmonary tuberculosis, and other specific respiratory disorders HIV-associated. Definitive diagnosis of PCP requires demonstration of Pneumocystis carinii (PC) in respiratory secretions or lung tissue. Conventional techniques, immunofluorescence using monoclonal antibodies and molecular techniques are highly specific, but sensitivity varies depending on the PC load present in the sample. Best diagnostic yield is obtained analyzing samples obtained by bronchoalveolar lavage. PC diagnosis using highly sensitive PCR in sputum-induced samples might allow noninvasive diagnosis in most HIV-infected patients suffering from PCP but PCR techniques remain to be standardized. Like in PCP prophylaxis, trimethoprim-sulphametoxazole (TS) is the drug of choice for PCP treatment. In severe case, TS is given intravenously. If patient is intolerant to TS, i.v. pentamidine or i.v. trimetrexate with folinic acid can be used. TS has a dose-dependent toxicity. In cases of hypersensitivity to TS, drug-desensitization should be tried. In severe documented PCP adjunctive corticosteroid therapy is effective and safe. In mild to moderate PCP, TS can be given orally. Best alternatives to TS in this situation are dapsone-pyrimethamine or clindamycin-primaquine (CP). Other effective options are oral atovaquone, aerosolize pentamidine and i.v. pentamidine. PMID- 9859619 TI - [Cerebral toxoplasmosis]. AB - Toxoplasmic encephalitis is associated with high mortality and morbidity and still presents a notable incidence in our setting. Neither the clinical symptoms nor radiological features are diagnostic of this disease; however, because of its frequency and clinical importance, specific treatment is begun whenever toxoplasmosis is suspected. In patients with negative serology, or who are receiving adequate prophylaxis, or who do not respond to 2 weeks of treatment, or who present radiological lesions suggestive of another illness, diagnosis should not be delayed, and brain biopsy should be considered as soon as possible. In these cases, SPECT with 201TI (sensitivity and specificity over 90-95% for lymphoma) and/or the PCR technique to detect T. gondii (sensitivity 50-65% and specificity 95-100%) or Epstein-Barr virus (sensitivity 70-80% and specificity 95% for lymphoma) can be very useful. The treatment of choice is pyrimethamine (100 mg the first day followed by 50 mg/day) and sulphadiazine (1-1.5 g/6 h) during 6-8 weeks. If the patient is allergic to sulfadiazine and cannot be desensitized the regimen of choice is pyrimethamine and clindamycin (600 mg/6 h), with similar efficacy. Clinical experience with other therapeutic alternatives is limited. Pyrimethamine can be associated with clarithromycin (0.5-1 g/12 h), azithromycin (1-1.5 g/day) atovaquone (750 mg/6 h), dapsone (100 mg/day) or doxycyclin (200 mg/12 h). Cotrimoxazole or clindamycin can be administered intravenously to patients who cannot receive enteral treatment. The toxicity of these therapeutic regimens is significant and treatment has to be suspended in 10 40% of cases. The interactions that can be produced with other drugs used to treat HIV-infected patients are generally of little clinical relevance. PMID- 9859620 TI - [Intestinal parasitic infections and leishmaniasis in patients with HIV infection]. AB - Intestinal parasite infections are very frequent in HIV patients with severe immunodeficiency (CD4 < 100/mm3) causing chronic diarrhea and malabsorption in the majority of cases. The most frequent microorganisms are microsporidia and Cryptosporidium parvum while Cyclospora cayetanensis and Isospora belli are more prevalent in subtropical and tropical areas and rare in industrialized areas. The diagnosis can be obtained by stool examination (differences in size and form of cysts), although microsporidia is frequently demonstrated by intestinal biopsy and/or duodenal aspirate. The treatment with cotrimoxazole for C. cayetanensis and I. belli is very effective and does not present any problems in the acute phase, however, due to a high percentage of relapses the treatment must be maintained while the patient is in a severe immunodeficiency state. E. intestinalis usually responds satisfactorily to albendazole while E. bieneusi is resistant to some drugs except in some cases (albendazole, atovaquone ad fumagillin). C parvum is also resistant to most medicaments but shows an adequate or partial clinical: response to paramomicine (< 50%). When there is no response, it is advised to administer octreotide since in half the cases the response is positive either total or partial. Nowadays with the use of protease inhibitors in the antiretroviral treatment a decrease in the incidence of these infections has been observed (microsporidia and C. parvum) even in the stools samples taken from the patients who had them before. As primary prophylaxis for C. parvum, it is better to avoid been exposed to the microorganism taking into account the 1997 preventive measures recommended by the USPHS/IDSA Prevention of Opportunistic Infections Working Group. The coinfection Leishmania-HIV is frequent in the mediterranean area. The most common specie is L. infantum. The incidence is most frequent in immunosuppressed patients (CD4 < 200 mm3) and in parenteral drug addicts. The symptomatology is similar to the one from immunocompetent patients, although in some cases it appears to be subclinical. A chronic development with relapses is frequent. The most effective diagnostic method for the finding of the parasites is thru bone marrow puncture and the culture in Novy-McNeal-Nicolle (NNN) medium. Serological tests have a low sensibility and the PCR is useful in asymptomatic cases, for therapeutical control and in relapses. The treatment is similar to that of immunocompetent patients, using primarily antimonials or amphotericine B (standard or lipid or liposomal forms). Relapses are very frequent, therefore, it is important to perform a secondary prophylaxis. However, no treatment has been completely effective. Mortality rate is high (approximately 25%) during the first month after diagnosis. This fact may be related to the severe immunodeficiency state and/or to the toxicity of the drugs used. The main priority for the future is to find a first line treatment with higher efficacy, decrease in relapses and a lower toxicity. PMID- 9859621 TI - [Other infections (Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, Salmonella spp., Campylobacter spp., Nocardia asteroides, Rhodococcus equi and Bartonella spp.)]. AB - People infected with human immunodeficiency virus (HIV) are at increased risk for bacterial infections due to HIV-associated immunologic defects. Bacterial infections were found to be, both a predictor of progression to AIDS and a substantial cause of mortality in pre-AIDS stages. Most bacterial infections are caused by Streptococcus pneumoniae, Haemophilus influenzae, Salmonella spp. and Pseudomonas aeruginosa. Rhodococcus equi, Nocardia spp., Campylobacter spp. and Bartonella spp. are less common. Data derived from two AIDS Clinical Trials Group studies showed that the most common bacterial infections were sinusitis (8.5 per 100 episodes per person years [py]), bacterial pneumonia (5.0 per 100 py), bronchitis (4.1 per 100 py) and soft tissue infections (3.5 per 100 py). In this review clinical characteristics and treatment recommendations according to data available in the literature for these infections are summarized. PMID- 9859623 TI - Emergency contraception: a modality whose time has come. PMID- 9859622 TI - [GESIDA survey on antiretroviral treatment in 1998. Working Group for the Study of AIDS (GESIDA) of the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC)]. PMID- 9859624 TI - Emergency contraception: a global overview. AB - Emergency contraception, sometimes referred to as "morning after" or postcoital contraception, provides a second chance for women who experience contraceptive failure or do not use a method, as well as for women who experience unplanned intercourse, including coerced sex or rape. The two primary methods of emergency contraception are postcoital use of a higher dose of oral contraceptive pills and insertion of an intrauterine device (IUD). Both can significantly reduce a woman's chance of becoming pregnant (75% and 99% respectively). Knowledge of emergency contraception is crucial, since women must know they can prevent pregnancy after intercourse in order to seek out treatment. While rates of unwanted pregnancy vary in different countries and among population groups, the need for emergency contraception is critical worldwide, However, the availability of emergency contraception differs widely. It is most extensively used in Europe, but is still a new method in other countries, including the United States. PMID- 9859625 TI - Safety of emergency contraception. AB - Emergency contraception (EC) prevents pregnancy. Four regimens are available in different parts of the world, a combination of ethinyl estradiol and levonorgestrel, levonorgestrel alone, mifepristone, and emergency insertion of an intrauterine device. All the regimens are also used either as long-term contraception or, in the case of mifepristone, as an abortifacient, and considerable data indicate their safety when used in these ways. Data on safety when the regimens are used as EC are lacking, but theoretically, and from practical experience, all appear to be extremely safe, particularly when compared to the risks of pregnancy. There has been a tendency to over-"medicalize" EC. Prescribing EC is simple. Consideration should be given to making EC available off prescription because it is so safe. PMID- 9859626 TI - Research on mifepristone and levonorgestrel in comparison with the Yuzpe regimen. AB - Women should be informed that it is possible to prevent unwanted pregnancy after intercourse in most cases by effective emergency contraception (EC). The currently used hormonal method, the Yuzpe regimen, however, has unpleasant side effects. The UNDP/UNFPA/WHO/World Bank Special Programme of Research, Development and Research training in Human Reproduction has worked on developing improved methods of EC for the last ten years. This research has focused on levonorgestrel and mifepristone. Now that results from large clinical studies are becoming available, they suggest that both these compounds are better tolerated and appear even more effective than the Yuzpe regimen. The challenge now is to implement the research results by making better emergency contraceptives a reality for women. PMID- 9859627 TI - The status of dedicated products. AB - Emergency contraception (EC) will not become a standard reproductive choice in the absence of dedicated products. Emergency contraception products based on the Yuzpe regimen have been available in Western Europe for a number of years. Levonorgestrel-only products are registered in 29 countries. Dedicated products of both types are being introduced into many developing countries and the United States. PMID- 9859628 TI - Should emergency contraceptive pills be available without prescription? AB - Leading health agencies, including the World Health Organization and the US Food and Drug Administration, consider emergency contraceptive pills (ECPs) safe and effective and have called for better access to them. Yet debate about whether ECPs should continue to be available by prescription only has been limited. After measuring the characteristics of ECPs against criteria developed to assess the necessity for prescription status for drugs generally, we argue that ECPs can safely be marketed over the counter. Professional assistance is not necessary since the woman diagnoses her own need for the pills and takes them herself. ECPs do not need to be adjusted for the individual woman and pose no potential threat of overdose or addiction. There are no contraindications except confirmed pregnancy (in which case the therapy will not work), and monitoring is not necessary. We conclude that prescription requirements that keep ECPs from women provide little, if any, benefit. PMID- 9859629 TI - When the morning after is Sunday: pharmacist prescribing of emergency contraceptive pills. AB - Pharmacists are well positioned to play a major role in increasing access to emergency contraceptive pills (ECPs). A pilot project in Washington State is testing direct pharmacist prescribing. Through a collaborative drug therapy agreement, a licensed prescriber, such as a physician, delegates to a pharmacist the authority to prescribe ECPs directly to women who meet the assessment criteria. Currently pharmacists at 111 Washington State pharmacies have collaborative agreements in place, and the number of participating pharmacies continues to increase. The response to this initiative has been extremely positive. Women who have received ECPs directly from pharmacists rate their interactions with the pharmacists positively and overwhelmingly cite convenience as the primary reason for going directly to the pharmacy. Physicians and other providers with independent prescribing authority can play a pivotal role by working with pharmacists to replicate the Washington State initiative in the states that allow it. PMID- 9859631 TI - Improving women's access to emergency contraception: innovative information and service delivery strategies. AB - Barriers to widespread use of emergency contraception (EC) include lack of knowledge on the part of women and providers, lack of support for the method from providers, and lack of a dedicated product in many countries. This article reviews strategies to improve women's access to EC launched by national or regional health authorities, clinicians, grass-roots health organizations, and women's groups. Information campaigns have targeted women to improve their knowledge of EC and providers to improve their comfort with it. Local groups and individual providers have also provided leaflets or designed innovative service strategies in order to improve women's access to the method. Expanding the scope or number of these programs and introducing them in areas where women do not currently have adequate knowledge of or access to EC will insure that more women will be able to use this method. PMID- 9859630 TI - Emergency contraceptive pills: what does the law say about prescribing, dispensing, repackaging, and advertising? AB - Despite the proven safety and efficacy of providing concentrated doses of oral contraceptive pills as postcoital contraception, many physicians and health care providers are unclear as to this "off-label" use and thus remain reluctant to dispense it. It is critical that health care providers learn about both the medical and legal aspects of providing emergency contraception so that they can make this important method of pregnancy prevention available to their patients. While a dedicated product may soon gain US Food and Drug Administration approval and go on the market, it is still important for health care providers to be aware of the legal issues surrounding the various aspects of both on- and off-label provision of emergency contraceptive pills. This paper provides an overview of the legal issues involved and suggests that there are a number of ways that health care providers can make emergency contraceptive pills available and readily accessible to patients with no serious legal risk. PMID- 9859632 TI - Are we making progress with emergency contraception? Recent findings on American adults and health professionals. AB - OBJECTIVES: To determine how awareness of and practices and attitudes toward emergency contraceptive pills (ECPs) have progressed among the American public and US health professionals. METHODS: In 1997, we conducted two nationally representative telephone surveys of Americans and health professionals of their knowledge, attitudes, and practices on ECPs and compared the findings to previous surveys. RESULTS: 66% of women and 51% of men 18 to 44 years old had heard of ECPs, up from 61% of women and 45% of men the same age in 1994. Only 1% of women surveyed reported having ever used this method, reflecting no change from 1994. Only 11% of women knew enough about ECPs to be able to use them. Americans named media as the primary source of information about ECPs. The proportion of physicians who had prescribed ECPs at least once in the preceding year increased significantly in 1997: 85% of obstetrician/gynecologists and 50% of family physicians compared to 69% and 34% in 1995. Almost all health professionals considered ECPs to be safe (99%) and effective (100%), yet relatively few discussed this option with their patients, and even fewer commonly prescribed it. CONCLUSION: Ongoing efforts are needed to improve awareness among the general public and to encourage health professionals to discuss and offer ECPs more widely. PMID- 9859633 TI - Call 1-888-NOT-2-LATE: promoting emergency contraception in the United States. AB - In 1997, the nonprofit Reproductive Health Technologies Project and the Office of Population Research at Princeton University, together with the communications firm Elgin DDB, planned and executed a mass media campaign to advertise the Emergency Contraception Hotline and more generally to further awareness of emergency contraception as a last chance means of pregnancy prevention in the United State. We produced a variety of public service announcements (PSAs) including television and radio spots in English and Spanish and several print versions adaptable for newspapers and magazines as well as outdoor settings such as billboards, transit shelters, and the sides of buses. Working with local coalitions, we succeeded in placing the PSAs free of charge in six pilot cities. We also generated coverage about the campaign in local and national news outlets. We chronicle the development of the media campaign, discuss the challenges and obstacles faced, and conclude with a review of the principal lessons learned. PMID- 9859634 TI - Emergency contraception: preliminary report of a demonstration and evaluation project. AB - Kaiser Permanente Southern California and the Pacific Institute for Women's Health began a demonstration and evaluation project on emergency contraceptive pills (ECPs) in the summer of 1996 with the goal of evaluating the feasibility and acceptability of ECPs in a large health maintenance organization and developing institutional templates, provider training and patient education materials that could be used to replicate the project. The ECP program had six components: repackaging of oral contraceptives in an ECP "kit," development of provider education materials, development of patient education materials, in service training, making ECPs kits available in convenient locations, and development of materials to support replication of the project inside and outside Kaiser Permanente. Although data are still being analyzed, preliminary results are promising. The success of the project within this relatively conservative, but well-established medical care organization provides a model for others. The development of a standard set of educational materials and approaches to implementation should facilitate dispensing ECPs in other settings. PMID- 9859635 TI - The impact of patient experience on practice: the acceptability of emergency contraceptive pills in inner-city clinics. AB - OBJECTIVES: This article reports on a study of 119 women who sought and used emergency contraceptive pills (ECPs) at Planned Parenthood of New York City (PPNYC) clinics between June 1996 and May 1997. It focuses on their satisfaction with the method, their attitudes toward ECPs, their reactions to the service, and the impact their perceptions had on changing the provision of care. METHOD: The PPNYC clinical protocol employed the Yuzpe method and fairly conservative procedures, including restrictive screening, a pelvic examination for all new patients, and limited appointment slots. A two-part survey captured information on patient experience with ECPs. RESULTS: The largest group of respondents (40%) found out about ECPs from friends. Almost 90% of the women were using contraception before their visit to the clinic. Sixty-eight percent reported that they sought ECPs because the condom failed. In the follow-up, a majority (57%) reported that they intended to change or had changed their method of contraception--more than three-quarters to a hormonal contraceptive. While generally satisfied with the service, many respondents were cautious of more extensive distribution of ECPs. CONCLUSION: The survey results had a profound impact on services: PPNYC revised the ECP protocol, developed a staff training package, expanded its service, and planned a multidimensional public media campaign. Further research, including a closer examination of participants' cautious attitude toward unrestricted distribution of ECPs, will be needed as PPNYC expands access to ECP. PMID- 9859636 TI - Inner-city adolescents' awareness of emergency contraception. AB - OBJECTIVES: To assess the awareness of emergency contraception (EC) among inner city adolescents attending a general primary health care clinic. METHOD: 197 patients filled out an anonymous 28-item survey on sexual activity, experience with contraceptives, attitude toward pregnancy, experience with pregnancy, awareness of and intent to use EC. RESULTS: 71% of the sample was sexually experienced; 90% had been active within six months of the clinic visit. While 81% of the sexually experienced segment of the sample had ever used contraceptives, 53% reported having had sex at least once during the past six months without using contraception. Fifty-seven percent "worried" following unprotected intercourse about a potential pregnancy; 32% of the sample had been involved in a pregnancy. Only 30% of the sexually experienced had heard of EC, but more than 87% stated they would use it if the need arose in the future. CONCLUSIONS: Urban adolescents are at high risk for unintended pregnancy due to inconsistent contraceptive use and/or method failure. Level of awareness of EC was low in our sample, particularly as compared to adult women in the United States, and to women of all ages (including teenagers) in European countries. Intent to use EC was high, however, indicating a strong desire to avoid unintended pregnancy. Attention should be focused on increasing both adolescent awareness of and access to EC. PMID- 9859638 TI - Emergency contraception among refugees and the displaced. AB - In 1994, the international relief community began to recognize and address the reproductive health needs of refugees and displaced populations. A minimum initial service package of reproductive health services for refugees and the displaced, which includes emergency contraception (EC), was developed and recommended for use in refugee settings. This paper describes the experience of one international relief organization, the International Rescue Committee (IRC), in introducing EC into its worldwide reproductive health program. A recent IRC survey found that EC is available in 4 out of 14 settings where it provides reproductive health services. A case study from Tanzania demonstrates the modes of delivery, the demand for EC by women who have experienced sexual violence, and the community responses to this method of contraception. More information, education, and communication directed at refugee communities; more donor support for supplies; and institutional commitment to train staff are needed to expand refugee access to EC. PMID- 9859637 TI - Emergency contraceptive pills: an exploratory study of knowledge and perceptions among Mexican women from both sides of the border. AB - A focus group study was conducted to explore what Mexican women from both sides of the US-Mexican border do to prevent pregnancy after unprotected intercourse, including their use of emergency contraceptive pills (ECPs); to examine their knowledge and perceptions of ECPs; and to explore what might influence Mexican women's use of ECPs. Eight focus groups (four in Mexico and four in California) were conducted with 55 sexually active Mexican women age 18 to 34. The women discussed a variety of postcoital methods intended to prevent pregnancy including herbs and injections. Although a third of the women had heard of ECPs, their knowledge was very limited. After being informed about the characteristics of ECPs, participants had many positive comments, preferring them to unplanned pregnancies and abortions. A major topic was whether or not ECPs are abortifacients. Having information about ECPs, a woman's personal circumstances, interpersonal factors, and the role of culture and religion were factors they thought would influence a woman's use of ECPs. After the focus group discussions, 95% of the women said they would use ECPs. Mexican women's acceptance and use of ECPs will likely depend on a host of factors within the broader context of their lives. PMID- 9859639 TI - [The effect of the acidity of the medium and of the temperature on the growth and polysaccharide excretion of Bacillus subtilis under submerged cultivation]. AB - Growth of Bacillus subtilis strains and excretion of polysaccharides have been studied under different environmental condition. These strains were studied as active antagonists of pathogenic microflora and were used for development of the biopreparation (probiotic), being efficient in treatment and prophylaxis of a number of bacterial infections of farm animals; the drug soon will be put into series production. The indices of accumulation of biomass of viable cells and exopolysaccharides (EPS) were used to study productivity of the strains. A possibility to increase yield in the production process was shown using the temperature factor and optimal pH of the culture medium under batch cultivation. The processes of the biomass and EPS accumulation were noncompetitive, almost coincided in time and were independent of the initial level of the pH of medium. PMID- 9859640 TI - [A search for site-specific type-II restriction endonucleases in streptomycetes]. AB - Three Streptomycetes strains producing endonucleases of restriction of II type have been found using the modified method of Belavin. PMID- 9859641 TI - [A comparative analysis of energy process inhibitors on the efficiency of phage infection in staphylococci]. AB - The study of the effect of KCN, DCCD and CCCP as inhibitors of the energy yielding processes showed that the efficacy of phage infection depended on respiration, proton ATPase, and proton electrochemical potential of hydrogen ions. There was a 49.5-68.0% decrease of the efficacy of phage infection after addition of the above mentioned inhibitors at the period of the contact of cells with bacteriophages at the stage of the phage nucleic acid transfer. The Embden Meyerhof-Parnas route inhibitors NaF and CH2ICOOH less affected the efficacy of phage infection. The same effect was observed during addition of Na3AsO4 as the ATP synthesis inhibitor. This efficacy decrease was probably due the inhibition of the processes of the substrate level phosphorylation and the deplete of the intracellular ATP content. PMID- 9859642 TI - [The cloning and expression of the gene for beta-galactosidase from Candida pseudotropicalis yeasts in Saccharomyces cerevisiae cells]. AB - The gene of beta-galactosidase of lactose-assimilating yeast Candida pseudotropicalis was cloned in pG2 and pBG2-3 hybrid shuttle vectors and expressed in Saccharomyces cerevisiae laboratory strains under the control of own promoter. The plasmids were able to replicate autonomously with relative stability in transformants of baker's yeasts. The availability of glucose or lactose in the medium influenced the recombinant plasmid stability and the expression of the cloned gene. A number of experiments have shown that the LAC+ phenotype in pG2-transformed Saccharomyces cerevisiae was due to the expression of the Candida pseudotropicalis lactose permease gene that is probably located in SaIG1/XhoI DNA fragment about 4.3 kb long. Southern hybridization experiments showed that LAC(+)-transformants of Saccharomyces cerevisiae contained both autonomously-replicative, and integrative pG2 plasmid. PMID- 9859643 TI - [The antibacterial properties of the intestinal microflora of healthy calves]. AB - Antibacterial properties of bacteria of genera Escherichia, Enterococcus, Bacillus, Lactobacillus, Bifidobacterium isolated from different parts of gastro intestinal tract of the healthy milk-fed and weanling calves were studied against test-bacteria, microorganisms of the Escherichia genus with and without pathogenic factors as well as the genera Klebsiella, Enterobacter, Proteus, Salmonella, Staphylococcus representatives that had been isolated from healthy and sick calves. Certain tolerance was determined between the groups of bacteria under study and only some strains of lactobacteria and bifidobacteria showed selective antibacterial action on pathogenic and conditionally pathogenic microorganisms, thus stabilizing intestinal microbiota in the gastro-enteric tract of calves. PMID- 9859644 TI - [The biological activity of a lipopolysaccharide from Ralstonia solanacearum ICMP 7859 and of its modified derivative]. AB - Coordinational compound of the Ralstonia solanacearum ICMP 7859 lipopolysaccharide (LPS) with germanium was obtained. The derivative has lost to considerable extent its toxicity (as compared to the initial substance) but preserves immunomodulating activity, interferon-inducing one in particular. Lipid A modification has led to complete loss of interferon-inducing activity. Apparently, phosphate at C4' GlcN II can be responsible for interferon-inducing activity of lipid A, while carboxylic groups of core oligosaccharide or 0 specific polysaccharide, are responsible for that in native molecule of LPS. PMID- 9859645 TI - [The characteristics of the causative agents of suppurative-inflammatory complications in hemophiliacs]. AB - Morphological-cultural and physiological-biochemical properties of 24 strains of microorganisms agents of pyo-inflammatory complications of different localization in patients with hemophilia have been studied. Microorganisms strains presented by the following species: Staphylococcus aureus, S. epidermidis, S. saprophyticus, Proteus vulgaris, P. morganii, Hafnia alvei, Serratia marcescens, have been identified. It was found out that in monoculture staphylococci prove to be the leading etiological agent (60.9%), gram-negative enterobacteria (52.2%) and bacterial associations (8.7%) occur more rarely. Special attention was paid to the study of resistance of antibiotics, circulation and pathogenicity factors that had a direct effect on the main disease severity. It was ascertained that high activity of enzymes and presence of pathogenicity factors were the peculiarities of microorganisms isolated from pyo-septic sites in patients with hemophilia. All the strains possessed multiple resistance to antibiotics. PMID- 9859646 TI - [Growth hormone in adolescents and in young adults]. PMID- 9859647 TI - [Management in an adolescent with Turner syndrome]. PMID- 9859648 TI - [Menstrual cycle]. PMID- 9859649 TI - [Contraception]. PMID- 9859650 TI - [Adolescents and medical services. Family pediatrics]. PMID- 9859651 TI - [Adolescents and health services: the hospital]. PMID- 9859652 TI - [Adolescents and health services: regional coordination]. PMID- 9859653 TI - [Adolescents and sexually transmitted diseases. Epidemiologic, preventive and social aspects]. PMID- 9859655 TI - [Vaccination of travelling and immigrant adolescents]. PMID- 9859654 TI - [Sexually transmitted diseases: laboratory diagnosis]. PMID- 9859656 TI - [Anti-hepatitis vaccination in children]. PMID- 9859657 TI - [Positive aspects of re-vaccination in adolescence]. PMID- 9859658 TI - [Idiopathic varicocele in adolescents]. PMID- 9859659 TI - [Spermatogenesis in developmental age]. PMID- 9859660 TI - [Medical assistance to adolescents in Europe]. PMID- 9859661 TI - [Sun and skin]. PMID- 9859662 TI - [Acne: therapeutic novelties]. PMID- 9859663 TI - [What adolescents eat]. PMID- 9859664 TI - [Sports and nutrients]. PMID- 9859665 TI - [Vitamin and mineral supplements in adolescents]. PMID- 9859666 TI - [The spermiogram in adolescence]. PMID- 9859667 TI - [Body changes in adolescence as a system of self-awareness]. PMID- 9859668 TI - [Optic neuritis and multiple sclerosis]. PMID- 9859669 TI - [A risk of developing multiple sclerosis following an outbreak of optic neuritis during a mean follow-up of 4.5 years]. AB - OBJECTIVE: The current study was designed to determine the risk of developing clinically definite multiple sclerosis (CDMS) after an isolated idiopathic optic neuritis (ON). PATIENTS AND METHODS: We retrospectively studied 28 patients (range from 18-45 years) who presented a unilateral acute ON between 1 st April and 31 st December. We excluded optic neuropathy of other causes, a previous diagnosis of MS or systemic diseases associated with ON. Patients underwent brain MRI, visual evoked potentials (VEPs), somatosensory evoked potentials (SEPs) and brainstem auditory evoked potentials (BAEPs). The mean interval between ON onset and MRI was 3.1 months. 24 patients were treated with corticosteroids in different ways: oral prednisone (14) and intravenous methylprednisolone (10). Mean duration of the follow-up was 4.5 years in 26 patients. RESULTS: Brain MRI detected white matter areas with increased signal in 10 of 25 patients (40%). Eight (30%) had bilateral anormalities on VEP, while SEPs and BAEPs revealed anormalities in one patient (5.5%). Two out 26 patients (7.7%) developed CDMS; one was treated with oral prednisone and the other with intravenous corticosteroids. No patients with normal MRI developed MS compared to 20% of patients with lesions on initial brain MRI. The only patient with abnormal SEPs and BAEPs did not develop MS. CONCLUSIONS: We found in our population low rate of developing CDMS with higher risk for those ON with abnormalities on initial brain MRI. PMID- 9859670 TI - [Familial episodic ataxia type 2. Clinical and genetic study of one family]. AB - INTRODUCTION: Episodic familial ataxia type 2 is caused by mutations in the gene CACNA1A, in chromosome 19p, that codifies part of a calcium channel. We report a family affected by this disease. PATIENTS AND METHODS: Nine members of this pedigree, all of them symptomatic, were seen and followed by us, including a magnetic resonance scan in all the cases but one. We performed linkage analysis to markers close or included in the gene CACNA1A. RESULTS: All of the patients had brief, self-limiting attacks of ataxia, that usually started between the ages of 8 and 12. Other symptoms frequently associated were dysarthria, headache, nausea and somnolence. Only one patient, with severe alcoholic intake, developed progressive ataxia after several years with self-limiting attacks. The remaining cases had nystagmus in lateral gaze as the only abnormality on examination. Acetazolamide decreased or eliminated the attacks in those patients treated. Magnetic resonance scans always showed cerebellar vermian atrophy. Genetic study confirmed linkage to gene CACNA1A. CONCLUSIONS: We emphasize the importance in knowing about this disease, with an easily identifiable clinical and neuroimaging pattern, and an efficient symptomatic treatment. PMID- 9859671 TI - [New applications of botulinum toxin]. PMID- 9859672 TI - [Alzheimer's disease and cellular membranes]. PMID- 9859673 TI - [Spontaneous epidural spinal hematoma]. PMID- 9859674 TI - [Spontaneous epidural spinal hematoma: report of 2 cases and review of the literature]. AB - Spontaneous spinal epidural hematoma is an uncommon cause of cord compression. This lesion has been correlated with some precipitant factors, but frequently is idiopathic. It's a unusual process, more uncommon in children, and there are only 350 cases reported in the literature. Etiology is unknown and there are several theories about it. The clinical presentation is remarkably uniform, with local back and radicular pain followed by sensory and motor disfunction. Early diagnosis and treatment are very important for the functional recovery of the patient. In this article, the literature is reviewed, especially the etiological and therapeutic aspects, and two new cases are reported. These cases can help to define the etiology of this entity. PMID- 9859675 TI - Extrapyramidal side effects and serotonin syndrome with fluoxetine and biperiden. PMID- 9859676 TI - [Striatal petechial hemorrhage and non-ketone hyperglycemia as the cause of hemichorea in a patient with unknown diabetes mellitus]. PMID- 9859677 TI - [Sensitive deficit of pseudo-polyneuritis distribution as the initial manifestation of spondylotic cervical myelopathy]. PMID- 9859678 TI - [Primary intraventricular hemorrhage secondary to postero-inferior cerebellar artery aneurysm]. PMID- 9859679 TI - [Words of misleading translation]. PMID- 9859680 TI - [Epidemiology of Parkinson disease]. PMID- 9859681 TI - [Current knowledge of the etiopathogenesis of Parkinson disease]. PMID- 9859682 TI - [Physiopathology of Parkinson disease]. PMID- 9859683 TI - [Parkinsonian syndromes and Parkinson's disease: differential clinical diagnosis]. PMID- 9859684 TI - [L-dopa: a fascinating history]. PMID- 9859685 TI - [Long-term complications of treatment with levodopa]. PMID- 9859686 TI - [Therapeutic strategies in advanced Parkinson's disease]. PMID- 9859687 TI - [Treatment of Parkinson's disease with COMT inhibitors: tolcapone]. PMID- 9859688 TI - Catechol-0-methyl transferase inhibitors in the treatment of Parkinson's disease. PMID- 9859689 TI - [Migraine and quality of life]. AB - The repercussion of migraine, as well as that of other chronic recurrent diseases, cannot be adequately evaluated with the classical epidemiologic indicators of morbidity and mortality. It is therefore necessary to use other parameters of evaluation such as the quality of life (QL) of the patients and the economic impact which they have on society. The authors herein review the concept of QL and the content and usefulness of the different tools which have been used to measure QL associated with health. The results of the QL studies in a general population are analyzed, compared with those for patients with primary headaches and other chronic diseases and with the differences among the different types of primary headaches, specially with the SF-36 questionnaire which is the most widely used measurement tool. All the specific questionnaires developed to evaluate the repercussion of migraine on the QL are also reviewed as are the results of its use in the different clinical trials, specially with the new drugs of the triptan family. We conclude emphasizing the importance of the measurement of QL in recognizing the socieconomic impact of migraine and recommending the use of these tools in the evaluation of new drugs since they new provide data patients' which cannot be obtained with the current efficacy measures. PMID- 9859690 TI - [Mechanism of action of zolmitriptan]. AB - Zolmitriptan is a new serotonergic agonist with excellent oral bioavailability exhibiting a potent symptomatic antimigraine effect. Zolmitriptan is a selective agonist of 5-HT1B/D receptors. 5-HT1B receptors are concentrated in the wall of the cranial extracerebral arteries. 5-HT1D receptors are located on the trigeminal terminals which receive pain from the leptomeningeal vessels. Migraine pain has its origin on cranial vessels. In fact, during a migraine attack the trigeminovascular system, which is composed by the cranial vessels and its trigeminal terminals, is activated. The activation of this system induces both dilatation and aseptic inflammation of cranial vessels. Zolmitriptan blocks both vascular phenomena. Its agonist action upon the 5-HT1D receptor ends the aseptic inflammation by inhibiting the release of vasoactive peptides. The dilatation of meningeal vessels disappears due to the stimulation of zolmitriptan of 5-HT1B receptors. As this drug crosses the blood brain barrier, zolmitriptan has both peripheral and central actions over the espinal trigeminal nucleus, which is rich in 5-HT1B/D receptors. Thus, the mechanism of action of zolmitriptan is double. On the one hand, zolmitriptan acts peripherally inhibiting dilatation and inflammation of cranial vessels. On the other, zolmitriptan exhibits a central nociceptive action in the brainstem nuclei. This dual action of zolmitriptan on migraine pain is completed with its beneficial effects on nausea and vomiting, due to its binding to the nucleus of the tractus solitarius, the center for control of vomiting. PMID- 9859691 TI - [Clinical efficacy of zolmitriptan in migraine]. AB - Zolmitriptan (previously known as 311C90) is a serotoninergic 5-HT1B/D agonist with high oral bioavailability with a double, central and peripheral, action mechanism. Evaluation of its clinical efficacy was developed in a program of clinical studies (search and confirmation of dosis, comparative and long term studies) and through analysis of efficacy in different clinical situations. Zolmitriptan shows a high effectiveness in the treatment of migraine crisis, significantly reduces the headaches by 2 hours of its administration, reduce the symptoms associated with migraine (nausea, photophobia and phonophobia) and improves the quality of life of the migraine patient. The efficacy is independent of the type of migraine characteristics of the patient as well as of the administration of other concomitant medications. The dosis of 2.5 mg of zolmitriptan has been found to be the optimum considering both efficacy and tolerability. PMID- 9859692 TI - [Safety profile of 311C90 (zolmitriptan)]. AB - A bibliographic review of the safety profile of 311C90 or zolmitriptan is performed in the present study showing the large number of clinical trials carried out in both healthy volunteers and patients with migraine. The molecule, a potent, selective agonist for the 5HT1B/1D receptors with central and peripheral activity does not appear to have significant influence on arterial pressure. ECG and Holter ECG studies did not show any alterations in healthy volunteers. In migraine patients, the ECG did not demonstrate ischemic alterations at any of the dosages of zolmitriptan used. In patients who had undertaken treatment for months, the hemogram and biochemical follow up did not show any changes. This new triptan was well tolerated in a wide spectrum of patients and healthy volunteers. Complaints of subjective side effects usually increase according to an increase in dosage. The most frequent adverse effects were nausea and dizziness. Other discomforts are: dryness of the mouth, sensation of heat, paresthesia, asthenia, drowsiness, and dizziness. The sensation of heaviness, tightness or pressure of the throat and chest have also been reported. The adverse effects reported with 5 mg of zolmitriptan are similar to those found with 100 mg of sumatriptan. The adverse side effects are usually mild, last a short time and remit without therapy. Zolmitriptan used together with the other most often used drugs in migraine patients did not show any important clinical interactions. However, it seems reasonable to limit the daily administration of zolmitriptan with monoaminoxidase inhibitors (MAOI-A) since a possible increase of the levels of zolmitriptan and its metabolites may be detected in the presence of one (moclobemide). At a dose of 2.5 mg, zolmitriptan appears to provide the best relationship between benefits and risk. PMID- 9859693 TI - [Treatment of mucosal leishmaniasis with aminosidine sulfate: results of two year follow-up]. AB - In 1996, 20 of 21 patients with mucosal leishmaniasis, treated in 1994 with aminosidine sulfate, 16mg/kg/day salt, by intramuscular injection for 20 days, were clinically evaluated. One patient died due to disease not related to mucosal leishmaniasis. Seven of 14 patients (66.7% N = 21) who achieved complete remission three months after treatment remained clinically cured 24 months later and seven relapsed in the same period (50% N = 14). Sorological follow-up showed poor correlation with the results of clinical examination. PMID- 9859694 TI - [Phlebotomines collected in remaining forests and wild animal shelters from the zoological garden of the Maringa urban area, southern Brazil. Preliminary study]. AB - The present study was undertaken after the detection of one case of cutaneous leishmaniasis with presumed infection in one of the three remaining wooded areas in the urban area of the city of Maringa, Southern Brazil; also in view of the lack of knowledge about sand flies and their behavior. From June to September, 1995, sand flies were caught with Falcao traps during the night in the remaining wooded areas (Parque do Inga, Bosque Dois and Horto Florestal). A total of 2,907 sand flies were caught in Parque do Inga; 1,723 of them were aught in forest traps and 1,184 in wild animal shelter traps at the zoo. The results show that Lutzomyia whitmani is better adapted to the three areas under study and that it frequently occurs in wild animal shelters within the urban perimeter of Maringa. PMID- 9859695 TI - [Killer toxin and enzyme production by Candida albicans isolated from buccal mucosa in patients with cancer]. AB - Opportunistic infections of the oral cavity are primarily caused by Candida and frequently occur in patients with cancer who are undergoing chemotherapy and antibiotic treatment. Of the specimens received from the oral mucosa of 44 patients with cancer, 25 (56.8%) yielded Candida on culture in Sabouraud agar. Twenty four of these isolates were identified as C. albicans (96%) and 1 as C. krusei (4%). The phenotypic characteristics of these isolates showed that all of them were strongly proteolytic, had a high ability to produce phospholipase, and presented the byotypes characterized as 811 (95.8%) and 511 (4.2%) in terms of susceptibility to killer toxins. PMID- 9859696 TI - [Contamination, by Toxocara spp eggs, in public parks and squares in Botucatu, Sao Paulo, Brazil]. AB - The frequency of Toxocara spp eggs in public parks was determined from March 1995 to February 1996. One hundred and twelve samples were collected from 10 public parks in Botucatu, Sao Paulo State, Brazil. Samples were processed by the decinormal sodium hydroxide concentration method. Out of the 120 soil samples analyzed, 21 were contaminated with Toxocara spp eggs, corresponding to a 17.5% rate of infestation. Of the ten squares submitted to analysis during the year, six presented contamination. Most of the ova found presented characteristics of inviability, with a consequent low chance of human infection, although the population is not free from the risk of contracting this zoonosis. PMID- 9859697 TI - Retrospective histopathological classification of 1,108 skin biopsies from patients clinically suspected of having leprosy from Bahia, northeast Brazil. AB - We report a retrospective histopathological classification carried out under laboratory conditions by the method of Ridley & Jopling of 1,108 skin biopsies from patients clinically suspected of having leprosy from Bahia, Northeast Brazil. PMID- 9859698 TI - [Histopathological and immunohistochemical characteristics of cutaneous and oral mucosa lesions in disseminated histoplasmosis associated with acquired immunodeficiency syndrome (AIDS)]. AB - Biopsy specimens of the skin and oral mucosa from twenty-five patients bearing the disseminated form of histoplasmosis (H. capsulatum) associated with AIDS (acquired immunodeficiency syndrome) were studied by histologic and immunohistochemistry techniques. Histologically, the skin lesions showed four different patterns: diffuse macrophage, granulomatous, vasculitic with leukocytoclastic and scarce inflammatory reaction. The cell markers for macrophages, lymphocytes B and T and H. capsulatum revealed CD68, UCHL-1 and L26 associated with variable amounts of fungi. PMID- 9859699 TI - [Utility of gamma rays in prophylaxis of transmissible malaria by blood transfusion]. AB - This study was carried out to evaluate the fortuitons advantage of using gamma irradiation in the prophylaxis of transmissible malaria by blood transfusion, with mice as the experimental model. In the first step, when the infected blood with Plasmodium berghei was submitted to 2,500 rad and 5,000 rad, with or without metronidazol, there was no success, because the animals presented parasitaemia and died after inoculation of irradiated blood. However, there was partial success in the second step, when the infected blood received 10,000 and 15,000 rad, and was inoculated in mice, which showed infection, and presented a survival rate of 20% and 40%, respectively, with later negativation of blood infected by P. berghei. PMID- 9859700 TI - [Strategy and perspectives for the control of cutaneous leishmaniasis in the state of Sao Paulo]. AB - The multiplicity of factors involved in the transmission of American cutaneous leishmaniasis (ACL) make it difficult to formulate an efficient strategy for the control of the disease. On the basis of the number of notified cases per location, the State of Sao Paulo determines the application of DDT in houses in those areas where two or more human cases have occurred. This procedure, however, impairs part of the program because 40% of the counties report only one human case each. These and other factors are analyzed in the light of the increase in the incidence of the disease in the State, its advance in the extra-forest environment and the delay in the decision-making process about the efficacy of the program. It is recommended that the control strategy be reviewed. PMID- 9859701 TI - [Coccidioidomycosis: a new brazilian case]. AB - A case of pulmonary coccidioidomycosis from the rural zone of Bertolinia, PI, is reported. The patient, a farm worker, attributed his illness to the dust inhaled while digging a water well during the dry season of the year, some weeks before the onset of the clinical manifestations. The main symptoms of the disease were severe chest pain and moderate fever. The diagnosis was made histopathologically: tissue phase fungal organisms--immature spherules and spherules with endospores- were observed in histological sections of a lung fragment obtained by open chest biopsy. This is the twelfth autochthonous case of coccidioidomycosis found so far in Brazil. All of them involved native inhabitants of the semi-arid part of Northeastern Brazil. The hot and dry environment of the region seems to favor the development of C. immitis in the soil. Humans and animals probably acquire the infection by digging the soil, when they become exposed to the conidium-bearing dust raised by this activity. PMID- 9859702 TI - [Acute/subacute disseminated paracoccidioidomycoses. First case in Rio Grande do Sul]. AB - The first autochthonous case of acute/subacute disseminated paracoccidioidomycosis observed in a child in Rio Grande do Sul (Brazil) is reported. The disease started with widespread superficial lymphadenopathy six months before the patient was admitted to the hospital. The diagnosis was made through a cervical lymph node biopsy. The spectrum of the clinical forms of the mycosis observed in this State is commented upon. PMID- 9859703 TI - [Progressive multifocal leukoencephalopathy as initial manifestation of acquired immunodeficiency]. AB - This is a report of a man with acquired immunodeficiency syndrome (AIDS) who presented acutely ill with severe progressive multifocal leukoencephalopathy (PML) as the first manifestation of AIDS. PML was diagnosed in the brain after gross and microscopical examination as well as by immunohistochemistry with an antibody against JC virus. PMID- 9859705 TI - Influence of Schistosoma mansoni infection on the reproductive capacity of albino mice. AB - This paper reports reduction on the reproductive capacity of female mice infected with Schistosoma mansoni, either in the acute phase or in the chronic one of the disease. This decrease in the reproductive capacity was highly significant (93.3% and 86.7%, for the acute and chronic phases, respectively). PMID- 9859706 TI - [Cost effectiveness of the treatment of HIV infection]. PMID- 9859704 TI - [American cutaneous leishmaniasis in a military training center located in Zona da Mata, Pernambuco, Brazil]. AB - An outbreak of American cutaneous leishmaniasis has been occurred in military training unit localized in 'Zona da Mata' of Pernambuco State, Brazil, where were registered 26 human cases. An epidemiological survey was carried out by entomological investigation and Montenegro skin test (MST). Lutzomyia choti presents predominance (89.9%) in sandflies identified. Out of 545 men who realized training activities were 24.12% positive to MST. PMID- 9859707 TI - [Infection and coronary atherosclerosis: the role Chlamydia pneumonia]. AB - The role of inflammatory reactions in the pathogenesis of atherosclerosis is widely accepted. Recently, an increasing body of evidence has linked infections to atherosclerosis. It is hypothesized that infections could interact with other risk factors of vascular disease, enhancing the endothelial damage and the production of atherosclerotic plaques. Several different infectious agents have been related to the atherosclerosis genesis: mainly herpesvirus, Helicobacter pylori and Chlamydia pneumoniae. Several lines of evidence strongly link C. pneumoniae to atherosclerosis. Consequently, several studies evaluating the effectiveness of antibiotic treatment in the reduction of cardiac ischemic events in patients with C. pneumoniae seropositivity have been performed. These studies support a causative role for C. pneumoniae. This article reviews the recent evidence linking infections to atherosclerosis, with emphasis on the role of C. pneumoniae on the atherosclerotic plaque. PMID- 9859708 TI - [Epidemiology of cardiovascular diseases in the Spanish elderly population]. AB - This paper is a comprehensive and critical review of the updated information available in Spain for the elderly population on the epidemiology of cardiovascular diseases. Clinical (coronary heart disease, heart failure, and cerebrovascular disease) and subclinical (left ventricular hypertrophy, carotid stenosis) cardiovascular diseases are reviewed. Prevalence and distribution of major classical cardiovascular risk factors such as hypertension, hyperlipidemia, diabetes mellitus and smoking and information on new risk factors such as microalbuminuria or abdominal obesity are also presented. The article is also focused on the high rates of morbidity, mortality and the burden of handicap in this age group in comparison with middle-aged people. Finally we call attention to the few and inconsistent population data available for some of the mentioned topics in our country, particularly the lack of specific figures of incidence and risk rates from cohort studies of elderly people in Spain. PMID- 9859709 TI - [Acute changes in wavelength of the process of auricular activation induced by stretching. Experimental study]. AB - OBJECTIVE: An evaluation is made of the acute modifications in the wavelength of the atrial excitation process induced by atrial stretching. MATERIAL AND METHODS: In 10 isolated Langendorff-perfused rabbit hearts and using a multiple electrode the wavelength of the atrial activation process (functional refractory period x conduction velocity) was determined in the right atrium. An analysis was also made of the inducibility of rapid repetitive atrial responses after 20 episodes of atrial burst pacing. Measurements were made under control conditions, after inducing two degrees of atrial wall stretch (D1 and D2), and following the suppression of atrial dilatation. RESULTS: Under control conditions the wavelength was 72.6 +/- 7.7 mm (250 ms cycle) and 54.0 +/- 5.1 mm (100 ms cycle). In D1 (mean longitudinal increase in atrial wall length = 24 +/- 3%) the wavelength shortened, with values of 59.8 +/- 6.6 mm (250 ms cycle; p < 0.01) and 44.9 +/- 5.1 mm (100 ms cycle; p < 0.01). In D2 (mean longitudinal increase in atrial wall length = 41 +/- 4%) the wavelength also shortened significantly, with values of 41.6 +/- 2.5 mm (250 ms cycle; p < 0.01 vs control) and 29.6 +/- 2.1 mm (100 ms cycle; p < 0.01 vs control). After suppressing atrial dilatation the wavelength was 65.7 +/- 8.0 mm (250 ms cycle, NS vs control) and 47.9 +/- 5.5 mm (100 ms cycle; NS vs control). The inducibility of rapid repetitive atrial responses increased during dilatation (22 episodes with over 30 consecutive repetitive responses in D1 [p < 0.01], 50 episodes in D2 [p < 0.001] vs 5 episodes under control conditions), and diminished after suppressing atrial dilatation (0 episodes with over 30 consecutive repetitive responses; p < 0.05). CONCLUSIONS: In the experimental model used, acute atrial dilatation produced a shortening in refractoriness and a decrease in conduction velocity. Both effects shortened the wavelength of the atrial activation process, facilitating the induction of atrial arrhythmias. The effects observed reverted upon suppressing atrial dilatation. PMID- 9859711 TI - [Therapeutic attitude in auricular fibrillation in emergency services]. PMID- 9859710 TI - [Retrospective study of auricular fibrillation of recent onset in a hospital emergency service: frequency, clinical presentation, and predictive factors of early conversion to sinus rhythm]. AB - AIM: To evaluate the proportion of emergencies due to recent-onset atrial fibrillation (AF), its clinical characteristics and in-hospital follow-up. The clinical predictors of conversion to sinus rhythm within the first 24 hours were analyzed. PATIENTS AND METHODS: 34,445 consecutive reports from patients presenting themselves at the emergency room of a community hospital during 15 months were retrospectively studied. The clinical reports of all patients (n = 186) with symptoms of recent onset AF (< 15 days) were reviewed. RESULTS: Hypertension (n = 77.41%) and lone AF (52 patients, 28%) were the most common etiologies. Forty seven patients (25%) presented with heart failure and the onset time was > 24 hours in 77 cases (41%). Conversion to sinus rhythm was observed in 71 out of 166 patients with at least 24 hours of follow-up (42.8%). Age < 60 years, the absence of cardiac disease, a NYHA functional class I, the absence of heart failure at the emergency room and the time from onset < 24 hours were significantly associated with conversion to sinus rhythm. The last two variables were selected as independent predictors by logistic regression analysis (sensitivity: 80%, specificity: 68%). CONCLUSIONS: We conclude that recent-onset AF represents 0.54% of all the hospital emergencies. The time from onset and the presence of heart failure predict the probability of conversion to sinus rhythm within the first 24 hours. PMID- 9859712 TI - [Cardiac manifestations of primary hypothyroidism. Determinant factors and treatment response]. AB - INTRODUCTION AND OBJECTIVES: Previous studies have not fully established the magnitude and determinant factors of cardiac manifestations of primary hypothyroidism. This study was aimed to assess the effects of thyroid deficiency on cardiac performance and structure. PATIENTS AND METHODS: We studied by echocardiography 19 patients with overt and 23 with subclinical hypothyroidism, and 21 control subjects. Patients were restudied one year after L-thyroxine therapy. Systolic function was assessed by the observed/predicted fractional shortening ratio. The predicted fractional shortening was calculated from the inverse relation of fractional shortening to end-systolic stress (p < 0.0001) in normal subjects. RESULTS: The observed/predicted fractional shortening ratio was lower (p = 0.043) and left ventricular mass was higher (p = 0.028) in overt hypothyroidism than in subclinical hypothyroidism and control subjects. By multivariate analysis, fractional shortening ratio was related to thyroxine levels (p = 0.0002), systemic vascular resistance (p = 0.0001) and age (p = 0.0009), and left ventricular mass was related to thyroxine levels (p = 0.0004) and weight (p = 0.0001). Pericardial effusion was observed in 37% of patients with overt hypothyroidism and 9% of patients with subclinical hypothyroidism (p = 0.03), and was mainly related to TSH levels (p = 0.0098). Hormone replacement therapy increased systolic function in overt hypothyroidism. Left ventricular mass did not change after therapy. Pericardial effusion disappeared in all patients. CONCLUSIONS: Primary hypothyroidism produces a decrease in myocardial contractility and an increase in left ventricular mass, both related to the severity of hormone deficiency. Pericardial effusion is mainly related to thyrotrophin plasma levels. Most of cardiac manifestations of hypothyroidism reverse with L-thyroxine therapy. PMID- 9859713 TI - [Maze technique for the treatment of auricular fibrillation: initial experience]. AB - INTRODUCTION AND OBJECTIVES: The MAZE procedure was developed as a surgical approach to the management of patients with atrial fibrillation refractory to medical treatment. This study seeks to identify the risk and benefits of adding the MAZE procedure in patients with atrial fibrillation undergoing surgery for underlying organic cardiac disorders. MATERIAL AND METHODS: Since november 1993, we have performed 10 interventions with the MAZE procedure, for the treatment of refractory atrial fibrillation. The indication to perform the technique was systemic embolism in 5 patients, contraindication for the anticoagulant treatment in two cases and no response to antiarrhythmic treatment in 5 cases. Two patients had more than one indication. In all the cases another surgical procedure was performed, 5 replacements of mitral valve, a mitral repair, one tricuspid repair and tree repairs of an atrial septal defect. RESULTS: Soon after surgery 9 patients were in sinus rhythm, and one in atrial fibrillation. Four patients needed atrial pacing during the first days. One patient required a pacemaker due to symptomatic sinus bradycardia. During the first 3 months, 4 patients had episodes of paroxysmal atrial fibrillation and flutter. One patient died suddenly one month after surgery. Seven patients have completed two years of follow-up, and are in stable sinus rhythm, in functional class I and free of antiarrhythmic drugs. All of them have echocardiographic evidence of mechanical activity in both atria. Left atrium had been reduced from 5.3 +/- 0.7 cm to 4.5 +/- 0.7 cm (p < 0.05). No patient has presented new embolic events. CONCLUSIONS: The MAZE procedure is a good choice in selected patients with atrial fibrillation refractory to medical treatment, or a precedent of systemic embolism. However, several problems can complicate the patient's course. PMID- 9859714 TI - [Complex implantation of an automatic defibrillator-cardioverter through a left superior vena cava]. AB - We describe the case of a 19-year-old girl with a left superior vena cava and a surgically corrected complete atrioventricular canal defect. After an inhospital sudden death an automatic defibrillator-cardioverter was implanted through her left superior vena cava. During the postoperative course, multiple inappropriate discharges caused by myopotential oversensing indicated the relocation of the electrode and, finally, insertion of two epicardial leads by a left submammarian thoracotomy approach, produced an optimal result. PMID- 9859715 TI - [Congenital anomalies of the mitral valve in the adult: presentation of 3 cases]. AB - Congenital mitral valve anomalies are uncommon and their incidence in adults is very unusual. Transthoracic echocardiography is essential for their diagnosis. In this study two adult patients with parachute mitral valve and one with isolated cleft of mitral valve are described. An echocardiographic exam was decisive in obtaining the diagnosis for each of these patients. Clinical aspects, typical echocardiographic findings and therapeutical approaches of these entities are discussed. PMID- 9859716 TI - [Refractory angina caused by steal phenomenon in internal mammary artery resolved with embolization]. AB - We report a patient with refractory angina in the postoperative period of a coronary artery bypass grafting. Ischemia was due to a large side branch of the left internal mammary artery causing steal phenomenon that was treated with transcatheter coil embolization. PMID- 9859717 TI - [Radiofrequency catheter ablation of supraventricular tachycardia in an adult with corrected tetralogy of Fallot]. AB - An 18 year old female with Fallot's tetralogy had undergone complete repair at thirteen years of age. Two years later she first presented a wide complex right bundle branch block tachycardia at a rate of 220 beats/min which could not be controlled on intravenous verapamil. Electrical shock successfully converted tachycardia to sinus rhythm, which showed typical Wolff-Parkinson-White syndrome. On electrophysiological study, the ortodromic tachycardia was found to be due to left lateral atrioventricular accessory pathway, which was ablated by radiofrequency catheter ablation. One year later she was symptom-free without antiarrhythmic medication. PMID- 9859718 TI - [Patent foramen ovale as a cause of recurrent cerebral embolism. Closure with minimally invasive surgery]. AB - We report a clinical case of a 63-year-old patient, referred with a history of repeated strokes, whose only cause was a patent foramen ovale with shunt right to left diagnosed by transesophageal echocardiography. He underwent minimally invasive surgery, through right parasternal minithoracotomy, practising defect closure. We comment on the value of echocardiography for diagnosing this pathology and the utility of this surgical technique for repairing these defects. PMID- 9859719 TI - [Clinical guidelines and prevention of ischemic heart disease in family practice]. PMID- 9859720 TI - [Serotonin uptake inhibitors (SSRI) in the treatment of children and adolescents]. PMID- 9859721 TI - [Visits by consultants and its effect on the clinical work and patient exchange. A Cochrane review of "outreach visits"]. PMID- 9859722 TI - [The auto- and endocrine function of the adipose tissue. Significance for metabolic complications in obesity]. AB - The present review discusses recent research showing adipose tissue to be highly metabolically active, producing and releasing many different bioactive compounds besides free fatty acids (FFA) such as tumor necrosis factor alpha (TNF alpha), leptin, acetylation stimulating protein (ASP), plasminogen activator inhibitor-1 (PAI-1), cholesterol ester transfer protein (CETP), prostaglandins and oestrogens. Most of these compounds have autocrine effects on the adipose cells and they are presumably involved in the physiological regulation of blood flow, growth and metabolism of the adipose tissue. When the adipose tissue becomes enlarged, as seen in association with obesity, it has now been shown that several of the compounds produced in the adipose tissue (TNF, PAI-1, CETP etc.) may be directly involved in the pathogenesis of some of the complications commonly seen in association with obesity such as insulin resistance, hypertension, enhanced thrombogenesis, and premature atherosclerosis. PMID- 9859723 TI - [The mini-mental state examination in screening of cognitive dysfunction and dementia]. AB - The mini-mental state examination (MMSE) is one of the most widely used screening instruments for the detection of cognitive impairments, used primarily in connection with screening for dementia. The intent of this review is to describe the original purpose of the MMSE and how it is currently used in clinical practice and in research. Advantages and limitations of the MMSE in providing a valid diagnosis of cognitive impairments (dementia) are discussed including sensitivity and specificity, the issues of the relationship of the MMSE scores to sociodemographic variables and examinations of factor structures of the MMSE. It is concluded that the MMSE provides a valid diagnosis of cognitive impairments among people with moderate and severe dementia in general populations. However, MMSE is not recommended as a screening instrument for the detection of early stages of dementia. Studies are needed to extend the present knowledge about how or whether the MMSE can be used in the clinical diagnostic evaluation of dementia and how demented patients treated with medications should be monitored. PMID- 9859724 TI - [Hand injuries in the users of hydraulic wood-cutters]. AB - This study has been carried out in order to suggest possible prophylactic recommendations. We investigated the type of injuries, the type of logsplitter used, the injury circumstances and the safety measures. Fifty-two patients were included. There were a total of 21 crushing injuries with amputations of fingers, hand, on forearm. Nine replantations, two revascularisations and seven amputations were carried out. Eighty-eight percent of the patients had their hand in the splitting area. Sixty-seven percent had not received any instructions on how to operate the machinery. In 58% of the cases two or more persons were operating the logsplitter and 40% of the machines had no emergency stop button. The hazards mentioned above influenced the injury frequence and severity. We therefore recommend improved prophylactic measures and precautions to be considered. For the old machines a campaign will be necessary, and for new logsplitters we recommend implementation of new standards. PMID- 9859725 TI - [Treatment of irritable colon with the food additive Appital]. AB - Appital is a herbal medicine. The aim was to examine its effect in patients with irritable bowel syndrome (IBS). Other diseases were excluded by physical examination, rectoscopy, blood tests and in patients older than 35 years X-ray of the colon or colonoscopy. The study was designed as a double-blind placebo controlled trial. The patients were randomized to either Appital or placebo. Following two weeks without medicine, the patient had Appital or placebo for eight weeks. Fifty-nine patients were randomized, 47 completed the study. The results were based on symptom scores registered by the patients. The symptom score was significantly reduced in patients treated with Appital (p = 0.002), but when compared to placebo, the difference was insignificant (p = 0.081). We concluded that Appital has no effect in relieving symptoms of IBS compared to placebo, although due to the possibility of type two error we cannot exclude a small, but hardly clinically relevant effect. PMID- 9859726 TI - [Adolescents and sexuality in family practice. A pilot study]. AB - It is well established that teenagers are generally dissatisfied with the sexual education they receive from school and parents. While several alternative sources of sex counselling have been suggested (i.e. an anonymous and semi-official telephone hot-line), the role of the general practitioner in sexual guidance of youngsters is only poorly evaluated. One hundred and sixty-eight teenage callers of "The Adolescent Sexuality Hot-line" were asked about their experience with sexual counselling by their family doctor. One third had actually discussed such topics with their g.p., and a total of 44.6% (significantly more girls than boys) regarded their g.p. as a desirable interlocutor on these issues. The reasons for having approached a g.p. for counselling differed according to gender, as did the causes for not wanting to involve a g.p. in sexual matters: boys tended to state shyness and lack of anonymity as main causes, whereas girls more often pointed to personal aspects of the doctor. PMID- 9859727 TI - [Seizures in acute apoplexy. Predisposing factors and significance for prognosis]. AB - The aim of the study was to determine the relationships between seizures during the early phase of stroke (early seizures, ES) and stroke outcome, and to identify predictors of ES. The study was prospective, consecutive and community based, and included 1197 patients with acute stroke. We determined the number and type of seizures, initial stroke severity, infarct size, mortality, and outcome in survivors. Stroke severity was measured on admission, weekly, and at discharge using the Scandinavian Stroke Scale (SSS). Multiple logistic and linear regression outcome analyses included relevant confounders and potential predictors. Fifty patients (4.2%) had seizures within 14 days of the stroke. In the multivariate analyses, only initial stroke severity was related to ES. For each 10-point increase in stroke severity (SSS score), the relative risk of ES increased by a factor of 1.65 (95% confidence interval, 1.4 to 1.9) (p < 0.0001). ES did not influence the risk of death during hospital stay (p = 0.56). In survivors, ES was related to a better outcome, equivalent to an improvement in SSS score of 5.7 points (SE [b] = 1.8; p = 0.002). The decisive factor of ES was initial stroke severity. ES per se was not related to mortality. Surprisingly, in survivors, ES predicted a better outcome. We explain this finding by a relatively larger ischaemic penumbra in patients who have ES after a stroke. PMID- 9859728 TI - [Visceral aneurysms. Two case reports]. AB - The authors report two case stories of visceral aneurysm: a 54 year-old man presenting recurrent gastrointestinal bleeding caused by a splenic artery aneurysm, and a 57 year-old man presenting with acute abdomen due to rupture of a hepatic artery aneurysm. Visceral aneurysms often present with classical gastrointestinal symptoms. They are connected with considerable mortality. Rupture requires instant surgery. In the elective phase the literature recommends embolization or resection. PMID- 9859729 TI - [Thymoma and pancytopenia]. AB - Thymomas are often associated with a variety of autoimmune disorders including various cytopenias. Rare cases have been associated with pancytopenia. We describe a patient with an invasive thymoma treated with thymectomy and subsequent radiation therapy. A few months later, rapidly progressing pancytopenia developed. Treatment with antibiotics, immunoglobulin, high-dose steroids and granulocyte-colony stimulating factor (G-CSF) had no effect, and the patient died within two months of overwhelming septicaemia complicated with intracranial haemorrhage. We review the literature and discuss therapies. PMID- 9859730 TI - [Intranodal hemorrhagic spindle cell tumor]. AB - We describe an example of a rare benign intranodal haemorrhagic spindle cell tumour (also called intranodal myofibroblastoma), occurring in a lymph node of the right inguinal region of a 53 year-old male patient. This is the first documentation of this tumour in the Danish literature. The lesion presents typically as a unilateral, solitary, painless inguinal lump. The microscopic appearance is characterized by proliferating spindle-shaped cells, interstitial haemorrhage and amianthoid fibers. Differential diagnosis includes primary and secondary lymph node tumours, such as Kaposi's sarcoma; metastatic spindle cell carcinoma; melanoma; neurilemmoma and soft tissue sarcomas. The clinical behaviour of the tumour is benign and local excision is the treatment of choice. PMID- 9859732 TI - [A reference program on hip fractures]. PMID- 9859731 TI - [Papillomavirus, uterine cervix cancer and screening]. PMID- 9859733 TI - [The death of Freud, again and again]. PMID- 9859734 TI - [Aamateur surgery]. PMID- 9859735 TI - [Indications for the use of serotonin uptake inhibitors in children and adolescents]. PMID- 9859736 TI - [Isotretinoin (Roaccutan) and depression]. PMID- 9859737 TI - [Withdrawal symptoms in connection with the use of selective serotonin reuptake inhibitors (SSRI)]. PMID- 9859738 TI - Stimulation of peripheral angiogenesis by vascular endothelial growth factor (VEGF). AB - Angiogenic growth factors constitutes a potentially novel form of therapy for patients with ischemic vascular disease. The feasibility of using recombinant formulations of angiogenic growth factors to augment collateral artery development by stimulation of capillary growth in animal models of myocardial and hindlimb ischemia has now been well established. In the case of vascular endothelial growth factor (VEGF) similar results may be achieved by gene transfer. Further laboratory and clinical studies may yield promising insights into the fundamental basis for native as well as therapeutic angiogenesis, and at the same time more explicitly define the manner in which therapeutic angiogenesis may be successfully incorporated into clinical practice. PMID- 9859739 TI - [Pathophysiology and clinical aspects of ischemia-reperfusion damage to skeletal muscles]. AB - Ischemia/reperfusion-injury of skeletal muscle--pathophysiology and clinical implications. Ischemia and reperfusion of skeletal muscle occurs in acute vascular occlusion and revascularisation, in elective vascular surgery, in orthopedic surgery by means of a tourniquet, and in transplantation of muscle containing cutaneous flaps. The ischemia sets up a cascade of events, which fully develops not prior to the moment of reperfusion. The ischemia-reperfusion-cascade is comparable to an overwhelming inflammatory reaction out of control. Depending on the duration of ischemia it prompts an injury for the muscle itself, and in some cases for the whole body. The present review discusses the main mechanisms of ischemia/reperfusion-injury in terms of cellular metabolism, endothelial function, cytokine release, and leucocyte function. Release of toxic oxygen radicals by activated leucocytes plays the pivotal role in this reaction. In addition, the clinical manifestations of ischemia/reperfusion will be reviewed as well as some of the means proposed to control this harmful reaction. PMID- 9859740 TI - Effects of endothelin and endothelin receptor antagonism in arteriolar and venolar microcirculation. AB - BACKGROUND: Endothelin-1 (ET-1) is a potent endogenous vasoconstrictor potentially involved in several cardiovascular diseases. The effect of ET-1 and the selective ETA receptor antagonist LU 135252 on skeletal muscle microcirculation in hypertensive rats was investigated. METHODS: The cremaster muscle of anaesthetised spontaneously hypertensive rats was superfused with 10( 8) M ET-1 with and without pre-treatment with LU 135252 10 and 30 mg/kg i.v. Vascular diameters were measured microscopically, recorded on videotape and quantified off-line. RESULTS: Superfusion with ET-1 led to a pronounced arteriolar constriction, which was the stronger the smaller the arterioles were (A1: 45%, A4: 90%). Venolar vasoconstriction was much less pronounced and independent of the vessel size (V1-V4: approx. 25%). LU 135252 (10 and 30 mg/kg i.v. was able to block arteriolar vasoconstriction dose-dependently, most pronouncedly so in the smallest arterioles. Venolar vasoconstriction was only antagonised by the higher dose. During the 30 minutes observation period cardiovascular parameters were not changed significantly with either dose of LU 135252. CONCLUSION: Selective ETA receptor blockade in hypertensive rats reduced ET-1 induced arteriolar vaso-constriction in resistance arterioles to a much higher degree then venolar constriction. As elevated ET-1 levels are seen in patients with primary hypertension, this new therapeutic principle may have promising clinical potential to treat hypertension by reducing peripheral arterial resistance. PMID- 9859741 TI - Rhythmical changes of the cutaneous blood flow in the forehead region under the condition of hypnoid relaxation. AB - BACKGROUND: A characteristic and stable blood flow rhythm can be detected for the skin of the forehead and ear lobes with frequencies of approx. 0.15 Hz (9/min), which were primarily not related to the respiratory rhythm. PATIENTS AND METHODS: The perfusion of the skin in the forehead region was investigated non-invasively with laser Doppler fluxmetry in ten healthy subjects before and during Hypnoid Relaxation (HyR). The HyR-state was induced by suggesting formulas regarding to the well known Autgeneous Training. RESULTS: In all test subjects rhythmical fluctuations of bloodflow with a frequency of approx. 0.15 Hz could be observed both, before and during HyR. We found that the amplitude of these fluctuations clearly (> 20% from individual baseline) increased in five of ten test subjects under the condition of HyR. Furthermore, in three of ten cases the spontaneous respiration under HyR adjusted to the frequency of the described bloodflow rhythm, which exists both, before and during HyR. CONCLUSIONS: These phenomena suggest an individually stabil and autonomous rhythm which is effected by alterations in the level of conciousness and which may be caused by the close linkage between the nerval structures for control of respiratory and circulatory systems. May be, this autonomic rhythm could be used as a trigger for breathing therapies or as a parameter for the impact of relaxation techniques on hemodynamics, e.g. in complementary therapy of vascular diseases like systemic sclerosis. PMID- 9859742 TI - Predicting healing of arterial leg ulcers by means of segmental systolic pressure measurements. AB - BACKGROUND: The Second European Consensus Document on Chronic Critical Leg Ischemia defined criteria to predict healing of leg ulcers. These recommendations are based on selected studies but the true performances of the tests proposed are not stated. METHODS: We performed an electronic and manual literature search to retrieve all studies that included the comparison of clinical outcome of cutaneous arterial leg ulcers with ankle (AP) and toe (TP) systolic pressures and we calculated the sensitivity and specificity of these measurements for predicting ulcer healing, according to standard methods for proportions and ROC curve analysis. RESULTS: Only three studies totalizing 220 legs with cutaneous ulcers (> 50% of the patients had diabetes) fulfilled our predefined criteria. The best performances of the tests (defined as the maximal area under the ROC curve) were obtained for an AP and a TP of 80 mmHg and 30 mmHg, respectively. Our analysis suggests that an AP > or = 80 mmHg predicts a favorable outcome with healing of leg ulcers. This simple test can be performed by the general practitioner. An AP of 50-80 mmHg should be followed by a TP and/or a transcutaneous PO2 measurement in case of unfavorable evolution. CONCLUSION: Our study confirms objectively the clinical validity of the criteria (AP < or = 50 mmHg, TP < or = 30 mmHg) supported by the Second European Consensus Document on Chronic Critical Leg Ischemia to predict unfavorable outcome of cutaneous arterial leg ulcers and provides objective performance values for these measurements. PMID- 9859743 TI - The hydrostatic measurement of systolic toe blood pressure: a preliminary validation of the method. AB - BACKGROUND: The hydrostatic method of measuring systolic toe blood pressure has not been validated. A comparison with established non-invasive methods is presented. PATIENTS AND METHODS: Conventional sphygmomanometry was applied to assess systolic arterial pressure at the ankle and brachial level; systolic arterial toe pressure was assessed by the hydrostatic method. An 8 MHz Doppler probe was used to demonstrate arterial pulsation in brachial, ankle, and toe arteries, respectively. 50 legs of patients with peripheral arterial occlusive disease were studied before and after successful arterial revascularisation. RESULTS: In all legs, ankle pressure and ankle/brachial pressure index (ABPI) increased postoperatively. Hydrostatic toe blood pressure (HSTBP) increased in 96% of cases, in 58% of cases above the upper detection limit of 70 mmHg. HSTBP and sphygmomanometric ankle pressures were significantly correlated (r = 0.754, p < 0.05). CONCLUSION: Successful arterial revascularisation in legs with peripheral arterial occlusive disease was associated with an increase in systolic ankle pressure and ABPI, assessed with sphygmomanometry. Similar changes were observed in systolic toe arterial pressure, assessed with the hydrostatic method. The correlation of the results obtained with both methods can be taken as a preliminary indication of the accuracy of the hydrostatic method, which may be particularly useful in cases of medial arterial calcification. PMID- 9859744 TI - Comparison in the interface pressure under self-adherent and non-self-adherent bandages during standing and exercise. AB - BACKGROUND: Self-adherent bandages are useful to prevent the bandage from becoming loose. However, the material binds tightly to itself to form a tight band, which may result in changes in interface pressure under the bandages during changes in posture and exercise, because the pressure depends on the elasticity of the bandages. The aim of this study was to compare the interface pressures under self-adherent and non-self adherent bandages while standing and during exercise. PATIENTS AND METHODS: In 20 normal volunteers the pressure was determined at the posteromedial area of the midcalf during lying, standing, tip toe exercise and walking, using an Air Pack Type Analyzer. We examined two kinds of bandages with a maximal tension of 70%, self-adherent and non-self-adherent bandages. RESULTS: There was no significant increase in pressure observed during standing in comparison with pressure in the supine position immediately after application. A significant increase in pressure under both bandages was observed during tip-toe exercise and walking. However, there was no significant difference in pressure during standing or exercise between self-adherent and non-self adherent bandages. Furthermore, there was no significant pressure difference between muscle contraction and relaxation between the two bandages. CONCLUSION: Self-adherent bandages can maintain their own elasticity, even when the material binds tightly to itself. PMID- 9859745 TI - Knitted double-velour Dacron prostheses in aortobifemoral position--long-term performance of different coating materials. AB - BACKGROUND: The purpose of this prospectively randomized study was to evaluate intraoperative as well as long-term performance of different coating materials for knitted Dacron aortobifemoral prostheses. PATIENTS AND METHODS: Between 1989 and 1992 a total of 150 consecutive patients who underwent aortobifemoral bypass were randomized for three different coating materials: gelatine (Unigraft; n = 49), collagen (Hemashield; n = 50) and human albumin (USCI; n = 51). Intra- and perioperative data such as duration of operation, clamping time, blood loss, and early complications were obtained as well as yearly follow-up examinations up to five years. Frequency of late complications, graft patency, and patient survival were observed. RESULTS: No statistically significant difference of intraoperative data could be obtained. In every group 6% of patients presented with anastomotic aneurysm in the groin. Primary patency rates were Unigraft 92%, Hemashield 91%, USCI 92% (no significant difference, n.s.), secondary patency rates after 5 years were Unigraft 98%, Hemashield 95%, USCI 94% (n.s.). CONCLUSIONS: Overall superiority of one particular type of prosthesis could not be stated. In order to prevent sudden graft failure associated with progressive arteriosclerotic disease or from late anastomotic aneurysm regular follow-up examinations should follow aorto-bifemoral bypass. PMID- 9859746 TI - Arterial reconstruction in diabetes and peripheral arterial occlusive disease: results in 192 patients. AB - OBJECTIVE: In the present study long-term patency and limb-salvage rates of femoro-distal bypasses were compared for patients with and without diabetes mellitus. METHODS: Between 1990 and 1994, some 192 femoro-crural bypass procedures were performed; 132 reconstructions were carried out on patients with peripheral arterial occlusive disease (PAOD), 60 bypasses on patients with PAOD and diabetes mellitus (DM). The two groups did not differ significantly regarding age, gender and risk pattern. RESULTS: The comparison of the two groups showed a significant advantage for the diabetic patients regarding the limb-salvage rate. In both groups there was also a significant difference regarding patency in favour of the diabetics. CONCLUSION: Patients suffering from diabetes and PAOD profit from femoro-distal bypasses. An aggressive vasosurgical reconstructive therapy is indicated for these patients. PMID- 9859747 TI - Successful thrombolysis in massive pulmonary embolism with right ventricular thrombus. AB - Systemic thrombolytic therapy for pulmonary embolism (PAE) is an established and common procedure. Due to increased risk, however, it is not much used in pulmonary embolism combined with thrombotic mass in the right ventricle. We applied it in a 59-year-old male patient with small intracardiac thrombus in the right ventricle (diameter = 1 cm) and peripheral pulmonary embolism whose diagnosis was obtained with transthoracal echocardiography (TTE) and helix lung CT. Twelve hours after thrombolytic treatment, helix lung CT scan showed a reduction in the size of the pulmonary embolism and no thrombotic masses in the right ventricle. In this patient with a small cardiac thrombus and rather peripheral pulmonary emboli, a systemic thrombolytic therapy proved to be effective and safe. PMID- 9859748 TI - Spinal cord stimulation in vibration white finger. AB - The authors report a case of vibration white finger syndrome in a 51-year old male, pneumatic drill worker. The patient complained of severe pain in the I, II, III and IV right fingers related to acral ischemic lesions. Dried skin with desquamation, tingling, paraesthesia and loss of sensation were present in both hands. Several arterial obstructions on forearm, hand and fingers were evident bilaterally at the angiography. Medical treatment, including administration of calcium-channel blockers, pentoxifylline and intravenous prostaglandin therapy, was unsuccessful. Under local anaesthesia an epidural spinal cord cervical electrode was implanted to control pain and ameliorate local microcirculatory conditions. The clinical result was excellent with the disappearance of symptoms and healing of acral lesions in a few weeks. Epidural spinal cord electrical stimulation represents an excellent technique for treatment in secondary Raynaud phenomenon related to vibration white finger syndrome. PMID- 9859749 TI - [Bypass to the anterior interosseous artery of the arm: follow-up of 3 years]. AB - Bypass to the anterior interosseous artery in the forearm: Follow-up at 3 years A 56 year-old, otherwise healthy woman suffered from acute ischaemia of her left arm one year after a fracture of the radius which had been treated with plaster fixation. Angiography demonstrated occlusions of the brachial, radial and ulnar arteries. Three surgical thrombectomies, local fibrinolysis and throacoscopic sympathectomy all failed to restore adequate blood supply to the forearm and hand. The patient continued to have pain on exercise and even at rest. After three months, an autologous vein bypass graft was implanted from the proximal brachial artery to the anterior interosseous artery. Thereafter, the patient became free of symptoms. At over three years, angiography was repeated and demonstrated a patent bypass. Furthermore, the lumen of the previously heavily diseased brachial artery had considerably improved in terms of diameter and regularity. These changes and possible mechanisms are discussed. PMID- 9859750 TI - A statistical consideration of CLAU-S: a disease specific questionnaire for the assessment of quality of life in patients with intermittent claudication. PMID- 9859751 TI - [Erwin Payr, Hans von Haberer, Ernst Jeger--a set of 3 stars in the universe of vascular surgery]. PMID- 9859752 TI - [The dynamics of the spatial organization of communities of small mammals (exemplified by rodents of the trans-Altai Gobi of Mongolia)]. AB - Between-year variation of rodent population densities was studied in the central part of trans-Altai Gobi, Mongolia, in summers of a nine year period. Variation in spatial structure of the desert rodent community in 1982-84 and in 1992 was examined by the use of discriminant function analysis. Although population densities, breadth and positions of niches of many species tended to vary independently, the positive conjunct variation was found for the most ecologically distinctive species. This can be explained by limitations of the climate. Although a guild structure of the community was pronounced each year, there were significant between year shifts in niche breadth and niche centres along certain axes of ecological space. Within year nearest neighbours in niche overlap (high seasonal overlap) differed in between year variation in abundance. It is supposed that interspecific competition could contribute to development of community structure. PMID- 9859753 TI - Precursors to pulmonary neoplasia. AB - Squamous dysplasia in the bronchi has been long recognized as a precursor of lung carcinoma, particularly squamous carcinoma. Atypical adenomatous hyperplasia (AAH) has been recently implicated as a precursor to adenocarcinoma. Bronchiolar neuroendocrine cell hyperplasia has also been suggested as a precursor to some pulmonary carcinoid tumors. The atypical adenomatous hyperplasia-adenocarcinoma sequence has been likened to the adenoma-carcinoma sequence in the large intestine. AAH is commonly multifocal, and may explain multicentricity that is observed with some adenocarcinomas. AAH has been shown to have immunohistochemical, morphometric, flow cytometric and genetic abnormalities overlapping with adenocarcinoma. Bronchiolar neuroendocrine cell hyperplasia (carcinoid tumorlets) is classically associated with inflammatory lesions in the airways, but may also be multifocal and bilateral. In the latter setting, lesions may attain a size greater than 0.5 cm and be (arbitrarily) classified as carcinoid tumors. PMID- 9859754 TI - Detection of human papillomaviruses in tissue specimens. AB - During the past decade, molecular methods based on the detection of viral DNA have become a key tool for the detection of human papillomaviruses (HPVs) in tissue. The methods can be divided into two groups: those in which tissue destruction is unavoidable for the detection of HPV DNA, and those in which the detection of viral DNA is performed in a way that allows tissue morphology preservation. Polymerase chain reaction is currently the most sensitive method for HPV detection and an excellent research tool. However, because of frequent contamination problems and lack of standardization, it is not readily applicable to diagnostic laboratories. The recent improvements in in situ hybridization have made it possible for this method to become the most appropriate method for routine detection of HPVs in tissue. At present, however, the use of at least two independent HPV DNA detection methods is indispensable for accurate determination of HPVs. PMID- 9859755 TI - Morphology, translocations, and clinical behavior: where do we go from here? PMID- 9859756 TI - Kuttner tumor (chronic sclerosing sialadenitis) of the submandibular gland: an underrecognized entity. PMID- 9859757 TI - Analysis of activation markers CD62 and CD63 after "in vitro" stimulation of platelets in antiphospholipid syndrome patients. PMID- 9859758 TI - Flow cytometry assay of phagocyte integrins in ischemic diseases. PMID- 9859759 TI - Flow cytometric analysis of fluorocytes in patients with erythropoietic porphyria. PMID- 9859760 TI - Quantitative analysis of P-glycoprotein, bcl-2 and transferrin receptor allows the stratification of acute myeloid leukemia patients within different prognostic risk classes. PMID- 9859761 TI - Megakaryocytopoiesis induced by PEG-rHu megakaryocyte growth and development factor (MGDF). PMID- 9859762 TI - The behaviour of blood lymphocyte subsets in patients treated with immunochemotherapy for metastatic melanoma. PMID- 9859763 TI - Detection of breast cancer cells using the magnetic cell separation (MACS) system: implications for stem cell purging. PMID- 9859764 TI - Monoclonal gammopathies with a high MC elicit altered phenotypic and genotypic features. PMID- 9859765 TI - Possible use of lymphoid CD subsets as prognostic markers for plasma cell dyscrasias. PMID- 9859766 TI - Comparison of clinical, morphological, immunophenotypical and cytochemical characteristics of LGL leukemia/lymphoma in dog, cat and human. PMID- 9859767 TI - Flow cytometry measurement of cytokine receptors in acute leukemias. Clinical and biologic implications. PMID- 9859768 TI - Diagnostic value of MPO and lysozyme antibodies in acute leukemia. Implications for the definition of FAB subtypes using a flow cytometric approach in combination with different permeabilising methods. PMID- 9859769 TI - Phenotypic changes of leukemic cells in a patient with prolympho-plasmacytic leukaemia after treatment with fludarabine. PMID- 9859770 TI - Lack of correlation between HLA-DR and CD25 "activation" related antigens on CD16+ NK cells in HIV infection. PMID- 9859771 TI - Surface P-glycoprotein expression on peripheral lymphocyte subpopulations from HIV-infected patients. PMID- 9859772 TI - Multicentric quality control trial for CD34+ cells evaluation in peripheral blood and leukapheresis collections from patients with hematological malignancies. Regione Piemonte Study Group. PMID- 9859773 TI - Comparison of four methods for CD34+ cell enumeration in cord blood. PMID- 9859774 TI - Simultaneous evaluation of reticulated elements in whole blood by flow cytometry. PMID- 9859775 TI - Over expression of alpha M beta 2 integrin on peripheral blood human neutrophils stimulated with GM-CSF. PMID- 9859776 TI - APO-1/Fas receptor (CD95) is non-functionally expressed in acute promyelocytic leukemias. PMID- 9859777 TI - Flow cytometric evaluation of activated eosinophils in graft versus host disease and hypereosinophilic conditions. PMID- 9859778 TI - Proliferative status and antigenic profile of CD34+ peripheral blood progenitors mobilized by chemotherapy and G-CSF. PMID- 9859779 TI - CD44 (H-CAM) is differently expressed on circulating and bone marrow hairy cells. PMID- 9859780 TI - Humoral-mediated suppression of lymphocyte blastogenesis in healthy donors receiving rhG-CSF. PMID- 9859781 TI - Expression of thrombospondin receptor (CD36) in chronic B-cell lymphoproliferative disorders: a role in tumor metastasis? PMID- 9859782 TI - CD44 protein in glioma rat cell culture. PMID- 9859783 TI - Cytofluorimetric evaluation of plasma cell proliferative activity in monoclonal gammopathies. PMID- 9859784 TI - Effect of antitumor platinum compounds with trans geometry on the induction of cytotoxicity and cell cycle modifications. PMID- 9859785 TI - Flow cytometric analysis of cell cycle phase perturbations induced by Thiocoraline, a new marine-derived anticancer compound. PMID- 9859786 TI - Antiphosphatidylserine antibodies in HIV-1+ patients bind apoptotic T cells in vitro. PMID- 9859788 TI - Taxol induces apoptosis in various cancer cell lines: comparison among different methodological approaches. PMID- 9859787 TI - Time- and dose-dependence of DNA fragmentation induced by anticancer agents: a flow cytometric study. PMID- 9859789 TI - Analysis of changes occurring in the cell cycle status of endometrial epithelial and stromal cells during the menstrual cycle by flow cytometry. PMID- 9859790 TI - Differences in cyclin B1 expression in cells blocked in the G2M phase after treatment with anticancer agent. New three-parametric flow cytometric analysis. PMID- 9859791 TI - Scavenger function of leukocytes phagocytosing apoptotic cells: analysis and quantitation by flow cytometry. PMID- 9859792 TI - Cell cycle kinetic effects and induction of apoptosis by all-trans retinoic acid (ATRA) in human MCF-7 breast cancer cells. PMID- 9859793 TI - Ki67 index and S-phase fraction on fine-needle aspirates from breast carcinomas. PMID- 9859794 TI - Detection of epirubicin (EPI) in human leukocytes and in plasma from ovarian cancer patients by flow cytometry. PMID- 9859795 TI - Measurement of antigen expression in early apoptotic cells. PMID- 9859796 TI - Apoptosis induced in glioma rat cells cultivated in the presence of a medicinal infusion of green tea. PMID- 9859797 TI - Life science applications of focused ion beams (FIB). PMID- 9859798 TI - An image analysis for the quantitative evaluation of K19 positive nodules in rat liver sections. PMID- 9859799 TI - Urine sediment analysis: comparison between microscopic evaluation and a fully automated flow cytometric analysis. PMID- 9859800 TI - Cell proliferation occurring during adult life is enhanced in vitamin E deficiency. PMID- 9859801 TI - Comparative analysis of wildtype and mutant forms of GFP. PMID- 9859802 TI - Three-dimensional reconstruction of FMRF-amide immunopositive neurons in the ventral ganglion of the barnacle Balanus amphitrite (Cirripedia, Crustacea). PMID- 9859803 TI - A computer-based technique for cell quantitation: "in vitro" chemotactic migration of endothelial cells. PMID- 9859804 TI - The use of CLSM to visualize the membrane flow during phagocytosis and exocytosis in Paramecium primaurelia. PMID- 9859805 TI - The flow cytometric cross match. Standardization of quantitative analysis. PMID- 9859806 TI - Expression and functional role of urokinase-type plasminogen activator receptor (UPA-R; CD87) in normal and acute leukemia cells. PMID- 9859808 TI - The effect of sample-to-sample variation on the estimation of cellular positivity from immunofluorescence test histograms. PMID- 9859807 TI - Flow cytometry assay of peripheral benzodiazepine receptors (pBZrs) on leukocytes following carbamazepine treatment. PMID- 9859809 TI - Discrimination of apoptotic cells in flow cytometry using trypan blue and FDA. PMID- 9859810 TI - Phenotypic heterogeneity of PHA-activated CD69+ lymphocytes: the expression of APO-1/FAS antigen (CD95) and perforins identifies different functional subsets. PMID- 9859811 TI - CD1b expression in Molt-4 clones exposed to IL-4 and GM-CSF. PMID- 9859812 TI - DNA ploidy, S phase fraction and telomerase activity in gastric cancer. PMID- 9859813 TI - Biological characterization of hydatidiform mole by flow cytometry. PMID- 9859814 TI - Correlation of cell proliferation and clinical behaviour of early gastric cancer. Immunohistochemical study with "in vivo" bromodeoxyuridine labelling. PMID- 9859815 TI - Biparametric analysis of DNA and Ag-NOR content for ploidy and kinetics. PMID- 9859816 TI - Ploidy and kinetics: comparative study with DNA flow cytometry and AgNOR protein quantification with image analysis. PMID- 9859817 TI - Bivariate analysis DNA/cytokeratin 7 and flow cytometric measurement of MIB-1 in superficial transitional carcinoma of the bladder (TCC). Methodological aspects and prognostic implications. PMID- 9859818 TI - Role of flow cytometric DNA analysis in the prognosis of endometrial carcinoma. PMID- 9859820 TI - P53 overexpression and DNA aneuploidy in colorectal adenocarcinomas. PMID- 9859819 TI - Flow cytometric S-phase fraction as an independent prognostic indicator in colorectal liver metastases. PMID- 9859821 TI - N-myc amplification and cell proliferation rate in human neuroblastoma. PMID- 9859822 TI - Hypothesis of clinical use of preoperative evaluation of DNA-index in endometrial carcinoma management. PMID- 9859823 TI - Increased yield of cancer cell detection in bronchial washing fluid combining conventional cytology and high-resolution flow cytometry. PMID- 9859824 TI - Retrospective analysis of the prognostic significance of DNA ploidy patterns in gastric cancer. PMID- 9859825 TI - Hepatocyte AgNOR protein quantitative expression is a powerful parameter for predicting hepatocellular carcinoma development in liver cirrhosis. PMID- 9859826 TI - Relationship between quantitative expression of protein p120 and proliferative activity in cancer cells. PMID- 9859827 TI - AgNOR protein expression and tumor growth rate of human carcinoma xenografts growing subcutaneously in nude mice. PMID- 9859828 TI - Monoclonal gammopathy and multiple myeloma: incidence of chromosomal aneuploidy detected by interphase fluorescence in situ hybridization. PMID- 9859829 TI - Aneusomy of chromosomes 1 and 17 in normal tissue adjacent to breast carcinomas. PMID- 9859830 TI - DNA flow cytometric analysis and numerical chromosome 9 aberrations detected by interphase cytogenetics in superficial bladder TCC. PMID- 9859831 TI - Ploidy analysis of paraffin-embedded breast fibroadenomas with flow cytometry and fluorescence in situ hybridization. PMID- 9859832 TI - In situ PCR (IS-PCR) to detect reduced amount of mouse minor satellite DNA. PMID- 9859833 TI - Combined cell sorting and FISH for detection of minimal residual disease in bone marrow of children with acute leukemia or solid tumors. PMID- 9859834 TI - Apoptosis and CD4+, CD8+ lymphocyte depletion following triphenyltin acetate (TPTA) exposure in mice thymic primary culture. PMID- 9859835 TI - Glutathione depletion enhances germ cell toxicity of butadiene metabolites: a flow cytometric study. PMID- 9859836 TI - Flow cytometric approach to study genome size variation in eurasiatic green toads of the Bufo viridis complex. PMID- 9859837 TI - Inter laboratory comparison for reliability and reproducibility of plant DNA flow cytometry. PMID- 9859838 TI - Senescence and 'rejuvenescence' in Colpoda inflata (Protista, Ciliophora): macronuclear DNA content and chromatin structure, evaluated by means of image analysis. PMID- 9859839 TI - Evolution of chromosome Y in primates. PMID- 9859840 TI - Lectin internalization and intracellular transport in Paramecium primaurelia cells blocked in phagocytic activity studied through CLSM. PMID- 9859841 TI - Human natural killer cell-target cell conjugate formation after exposure to extremely low frequency magnetic fields: a cytometric analysis. PMID- 9859842 TI - Rapid flow cytometric immunodetection of bacteria to monitor aquatic environments. PMID- 9859843 TI - Pathways of cadmium uptake in excitable mammalian cells: a microspectrofluorimetric study. PMID- 9859844 TI - Flow cytometric evaluation of CD38, CD45 RO and CD45 RA in HIV-infected children. PMID- 9859845 TI - Detection of P-glycoprotein efflux activity on peripheral blood lymphocyte subsets from HIV+ patients. PMID- 9859846 TI - CD16+CD56+CD8+ natural killer (NK) cells are decreased during HIV infection. PMID- 9859847 TI - Detection of the cytomegalovirus antigen in the neutrophils of peripheral blood by flow cytometry. PMID- 9859848 TI - Programmed cell death in children with active visceral leishmaniasis. PMID- 9859849 TI - alpha-Gliadin-induced T cell activation from the peripheral blood measured by CD69 expression. PMID- 9859850 TI - A national policentric study for the definition of a reference range for normal values of peripheral blood lymphocyte subsets in healthy adults. PMID- 9859851 TI - Toward a biobehavioral model of visceral hypersensitivity in irritable bowel syndrome. PMID- 9859852 TI - Experimental pathophysiology of panic. AB - In this article, we review how the knowledge of the pathophysiology of panic disorder has expanded, with special emphasis on laboratory models using lactate and carbon dioxide challenges. Experiments in the late 1960s revealed that lactate infusion can induce panic attacks. A prominent feature of these attacks is hyperventilation. Because lactate infusion induces a metabolic alkalosis, one would rather expect a compensatory hypoventilation. For years hyperventilation was thought to be causally linked to panic, but it has since been proven to be a symptom rather than a cause of panic attacks. Similarly, it is not hypocapnia but hypercapnia that has proven to be capable of provoking panic attacks. Carbon dioxide challenges are comparable to lactate infusion in the degree to which they meet the criteria for an ideal model of panic disorder. Experiments with carbon dioxide in first-degree relatives of panic disorder patients and in monozygotic twins support the idea of a constitutional predisposition to panic disorder. Of the various other agents that have been used to trigger panic attacks, cholecystokinin seems particularly promising as a valid laboratory model of panic disorder and may provide valuable data regarding the mechanism of panic attacks. The false suffocation alarm theory, proposed by Klein, is an integrative hypothesis that may account for a large number of the laboratory as well as clinical observations. PMID- 9859853 TI - The persistence of fatigue in chronic fatigue syndrome and multiple sclerosis: development of a model. AB - The cause of chronic fatigue syndrome (CFS) is unknown. With respect to factors perpetuating fatigue, on the other hand, a model has been postulated in the literature in which behavioral, cognitive, and affective factors play a role in perpetuating fatigue. In the present study, this hypothesized model was tested on patients with CFS and on fatigued patients with multiple sclerosis (MS). The model was formulated in terms of cause-and-effect relationships and an integral test of this model was performed by the statistical technique, "structural equation modeling," in 51 patients with chronic fatigue syndrome and 50 patients with multiple sclerosis matched for age, gender, and education. Attributing complaints to a somatic cause produced low levels of physical activity, which in turn had a causal effect on fatigue severity. Depression had to be deleted from the model. Sense of control over symptoms and focusing on bodily symptoms each had a direct causal effect on fatigue. The model showed an excellent fit for CFS patients, but was rejected for MS patients. Therefore, a new model for MS patients had to be developed in which sense of control had a causal effect on fatigue. In the MS model, no causal relationship was found between the physical state as measured by the Expanded Disability Status Score (EDSS) and fatigue or functional impairment. The present study shows that cognitive and behavioral factors are involved in the persistence of fatigue. Treatment should be directed at these factors. The processes involved in the subjective experience of fatigue in CFS were different from the processes related to fatigue in MS. PMID- 9859854 TI - Subjective symptoms and breathing pattern at rest and following hyperventilation in anxiety and somatoform disorders. AB - The purpose of the present study was to investigate the diagnostic specificity of bodily symptoms and respiratory behavior at rest and after a hyperventilation provocation test (HVPT) in patients that were either grouped according to the DSM classification or diagnosed as suffering from hyperventilation syndrome. Nine hundred three anxiety and somatoform patients, showing symptoms supposedly caused by psychogenic hyperventilation, and 170 healthy subjects, were studied. Breathing pattern and end-tidal CO2 concentration were recorded during breathing at rest and following a HVPT. Subjective symptoms in daily life and after HVPT were measured. A principal-components analysis was performed on both the symptoms and breathing variables and their specificity levels were compared in the two classifications of patients. Some symptoms in daily life were grouped together with the same symptoms after the HVPT, other symptoms were not. This suggests that the HVPT elicited partly specific symptoms, and partly reproduced the symptoms experienced in daily life. Similar findings were observed with respect to the breathing variables. Patients with panic differed from other patients with anxiety disorders by an increased level of symptoms and a FETCO2 decline at rest. The HVPT may be informative for diagnosis because it provokes some of the typical somatic and psychological symptoms, and it identifies the breathing instability that is characteristic of both patients with HVS and with anxiety. The same symptoms and breathing variables characterized the patients, whatever their classification. Overall, the specificity of breathing variables is rather low. PMID- 9859855 TI - The changing profile of anorexia nervosa at the Toronto Programme for Eating Disorders. AB - The present study is a retrospective comparison of anorexic patients of the restricting subtype (AN) with those of the bulimic subtype (AN+BN) over three consecutive 5-year time periods. Subjects consisted of 806 female out-patients diagnosed as either AN or AN+BN between 1978 and 1994 during an assessment of their eating problem at the University of Toronto Programme for Eating Disorders in Toronto, Canada. During each time period an increasing proportion of subjects was diagnosed with AN+BN than with AN. Subjects with AN+BN reported a longer duration of illness prior to being assessed. Over time, subjects in both groups appeared to weigh more, and were less likely to report amenorrhea. Subjects from both groups reported higher frequencies of purging behavior, impulsive behaviors, and associated affective symptoms in the later time periods. Over time, the differences between the two subgroups of patients became less distinct. The results indicate changes in the clinical profile of patients with anorexia nervosa seen at the Toronto Programme for Eating Disorders over the past 16 years. The implications of these changes are explored. PMID- 9859856 TI - The validity of the Dutch Sleep Disorders Questionnaire (SDQ). AB - The Sleep Disorders Questionnaire (SDQ) is a 176-item questionnaire designed to diagnose the presence of common sleep disorders. This study set out to assess the validity of a Dutch translation of the SDQ. Scores on 145 questionnaires were analyzed. A cluster analysis of these scores revealed the following clusters: healthy, depression, insomnia, narcolepsy, and apnea. The cluster classification proved correct for 67% of the subjects, as determined on the basis of polysomnography. These results show that the Dutch SDQ is a reasonably valid instrument for diagnosing sleep disorders. PMID- 9859857 TI - Salivary cortisol in women with and without perimenstrual mood changes. AB - An increase in the activity of the hypothalamic-pituitary-adrenal axis (HPA axis) is frequently associated with major depression. During the premenstrual phase of their reproductive cycle some women experience depressive mood changes that are proposed to be of similar intensity to that experienced during periods of major depression. This study examined the secretion of cortisol, the end-product in the HPA axis, at different stages of the menstrual cycle in women with and without premenstrual depression. Women who experienced only mild physical and emotional changes in the premenstrual phase of their cycle had a significantly higher cortisol secretion on a premenstrual day (measured hourly) compared to a postmenstrual day. Those who were significantly more depressed premenstrually showed the opposite pattern of cortisol secretion with significantly lower levels on the premenstrual day compared with the postmenstrual day. Across the menstrual cycle, women who were significantly more depressed premenstrually also had lower evening cortisol levels in their premenstrual phase. The results of this study indicate that, unlike major depression where the underlying neurological changes are manifest as overactivity of the HPA axis, premenstrual depressive changes are associated with reduced HPA axis activity. Premenstrual depression may, therefore, be similar neurologically to seasonal affective disorder, which is associated with underactivity of the HPA axis. PMID- 9859858 TI - Identification of differentially expressed genes in human trophoblast cells by differential-display RT-PCR. AB - Molecular mechanisms controlling human trophoblast invasiveness are still poorly understood. In the present investigation, mRNA patterns of trophoblast cells isolated from first trimester (high invasiveness) and term placentae (no/low invasiveness) were compared by differential-display reverse transcriptase polymerase chain reaction (DDRT-PCR) revealing differential expression of numerous genes. Of 18 differentially expressed DDRT-PCR products analysed, 11 were unknown, four showed homologies with expressed sequence tag sequences, and three others were homologous to integrin-beta1, ATP-synthetase U6 (both showing higher expression in first trimester) or to aldose-reductase (higher expression at term), respectively. One of the unknown transcripts (PBK1, accession number: AJ007398) was cloned from a first trimester placenta cDNA library and was characterized. The 1908-bp gene fragment contains an open reading frame of 1551 bp and an Alu-sequence in the 3' non-coding region. According to Northern blot analysis on JAr choriocarcinoma cells, the fragment is close to full-length cDNA. By in situ hybridization, PBK1 was detected only in first trimester but not term placentae in the proximal parts of cell islands and in closely adjacent villous cytotrophoblast. This expression pattern suggests that the newly identified molecule, PBK1, could be involved in the regulation of proliferation/ differentiation and potentially in invasion of trophoblast cells. PMID- 9859859 TI - Interaction of interstitial trophoblast with placental bed capillaries and venules of normotensive and pre-eclamptic pregnancies. AB - While endovascular trophoblast invasion of the human placental bed spiral arteries has been studied extensively, no information is available on the interaction between interstitially invading trophoblast and uterine capillaries and venules. Placental bed biopsies of eight normotensive and 15 pre-eclamptic patients were double-immunostained for cytokeratin and the endothelial marker CD31, providing satisfactory staining results in six and 10 biopsies, respectively. Interstitial trophoblast tissue density did not differ between the two series of biopsies, implying that this pathway of invasion is not impaired in pre-eclampsia. Both groups showed a similar incidence of approach of non-arterial vascular structures by perivascular trophoblast. Differences in CD31 staining intensity were noticed in different vascular cross-sections. Lower staining intensity was related to the presence of perivascular trophoblast. Because of the identity of CD31 with the platelet-endothelial cell adhesion molecule (PECAM)-1, the trophoblast-dependent downregulation of CD31 may play a role in the control of leukocytic traffic within the placental bed. The phenomena described in this paper did not show any difference between the normotensive and pre-eclamptic patients, implying that interaction of interstitial trophoblast with venous and capillary structures is not related to the pathogenesis of pre-eclampsia. PMID- 9859860 TI - Pregnancies complicated by retained placenta: sex ratio and relation to pre eclampsia. AB - Pre-eclampsia and placenta accreta have opposite histological features of placentation. This study set out to test the hypotheses that the sex ratios in these two pregnancy complications are opposite and that these conditions are mutually exclusive. A population-based database covering all deliveries in South Australia between 1986 and 1995 and the hospital-based obstetric database of the Adelaide Women's and Children's Hospital, covering 8549 births between 1993 and 1995, were used to ascertain the sex ratios in singleton pregnancies and the sex ratios in those pregnancies in which there was retained placenta, hypertension in pregnancy, or pre-eclampsia. The likelihood of independence of occurrence or mutual exclusivity of retained placenta and hypertension in pregnancy or pre eclampsia were also examined. The male:female sex ratio in the South Australian population was 1.077. In pregnancies with hypertension in pregnancy it was 1.165 (P<0.001) and in pregnancies with retained placenta it was 0.883 (P<0.0001). There was a trend to an increased sex ratio in pre-eclamptic pregnancy (1.248 in primigravid and 1.092 in multigravid women) but there was insufficient power to detect significance (P=0.207 and 0.470, respectively). Neither hypertension in pregnancy nor pre-eclampsia were mutually exclusive of placenta accreta: hypertensive disorders of pregnancy and placenta accreta occurred independently of each other. Our findings suggest that sex-linked antigens are unlikely to influence maternofetal interactions consistently to give rise to one but not the other pregnancy complication. PMID- 9859861 TI - Placental mitochondria as a source of oxidative stress in pre-eclampsia. AB - Pre-eclampsia is a hypertensive disorder of human pregnancy that is a leading cause of premature delivery and fetal growth retardation. It is characterized by hypertension, reduced uteroplacental blood flow, proteinuria and oedema. Pre eclampsia is associated with increased lipid peroxidation in the maternal circulation and in the placenta. Mitochondria are sources of oxygen radicals and are enriched with polyunsaturated fatty acids that are susceptible to peroxidation. Therefore, the mitochondria could be an important source of oxidative stress and lipid peroxidation. To study this, the level of lipid peroxidation in the mitochondrial fraction of placentae obtained from normally pregnant women (n=8) and women with pre-eclampsia (n=8) was examined. Placental tissues were homogenized and the mitochondrial fraction was isolated by ultracentrifugation. Mitochondrial lipid peroxides were estimated by malondialdehyde (MDA). NADPH and Fe++ were used to stimulate lipid peroxidation. Superoxide dismutase (SOD) was used to inhibit superoxide radicals and mannitol to inhibit hydroxyl radicals. The following results were found: (1) MDA levels were significantly greater in the mitochondrial fraction isolated from pre eclamptic placentae than from normal placentae (27.4+/-3.0 versus 17.0+/-1.8 nmol/g tissue, mean+/-s.e., P<0.05); (2) the oxidative potential of the pre eclamptic mitochondrial fraction was also higher than normal as evidenced by the significantly greater stimulation of lipid peroxidation by NADPH and Fe+ + (248+/ 25 versus 164+/-35 nmol/g, P<0.05); (3) superoxide dismutase, but not mannitol, attenuated the lipid peroxidation induced by NADPH and Fe+ + demonstrating that superoxide is the radical responsible for mitochondrial lipid peroxidation in this system; and (4) the amount of mitochondrial protein was 47 per cent greater and the activity of the mitochondrial enzyme, citrate synthase, was 56 per cent greater in the pre-eclamptic placentae indicating an increase in the amount of mitochondria in the pre-eclamptic placentae. It is concluded that: (1) mitochondrial lipid peroxidation is increased in pre-eclampsia; (2) the amount of placental mitochondria is increased in pre-eclampsia; (3) placental mitochondria contribute to the abnormal increase in lipid peroxidation that occurs in pre eclamptic placentae by both an increase in their amount and an increase in their susceptibility to oxidation; and (4) mitochondrial generation of superoxide could be an important source of oxidative stress in pre-eclampsia. PMID- 9859863 TI - Intrauterine expression of parathyroid hormone-related protein in normal and pre eclamptic pregnancies. AB - Maternal hypertension, vasoconstriction and placental insufficiency are features of pre-eclampsia. Alterations in calcium homeostasis and in the production of calciotropic hormones and vasoactive agents have also been described in association with pre-eclampsia. Parathyroid hormone-related protein (PTHrP) is abundantly expressed in intrauterine tissues during normal pregnancy and has roles in fetal growth and calcium homeostasis, placental calcium transport and vascular tone regulation. Intrauterine PTHrP mRNA expression and tissue PTHrP content were determined by Northern blot analysis and radio-immunoassay, respectively, in preterm and term pre-eclamptic women. PTHrP mRNA expression and PTHrP content in placenta, amnion over placenta, reflected amnion and choriodecidua from preterm pre-eclamptic women (n=8-10) were not different from preterm controls (n= 10-12). PTHrP mRNA expression and content in amnion over placenta and reflected amnion were significantly greater in term compared to preterm pre-eclamptics (P<0.05). PTHrP mRNA expression was significantly lower in choriodecidua from term pre-eclamptic women (n=8) compared to term controls (n=28, P<0.05), but was not different in placenta or amnion. PTHrP content was not altered in term pre-eclamptic women (n=8) compared to controls (n=25) for any tissue. In summary, PTHrP expression in placenta and amnion was not increased in pre-eclamptic women in association with maternal hypertension, placental insufficiency and vasoconstriction. PTHrP mRNA expression was decreased in choriodecidua in association with term but not preterm pre-eclampsia, however, levels of the protein were not decreased. The data suggest that PTHrP is not involved in the placental pathophysiology of pre-eclampsia in late gestation. PMID- 9859862 TI - Invasion of cytotrophoblastic JEG-3 cells is stimulated by hCG in vitro. AB - Trophoblast invasion into the uterine wall is controlled by many factors. Previously, a human chorionic gonadotropin (hCG) receptor has been found to be expressed on invasive trophoblast as well as on choriocarcinoma cells implying a possible role for the hormone in trophoblast invasion. Therefore, this study examined the role of hCG in the invasion of trophoblastic (JEG-3) cells. Increasing hCG concentrations were applied in a trophoblast invasion model, JEG 3, through matrigel-coated filters. The proliferation was quantified by WST-1 cleavage assay. Cell migration was studied by examining the number of cells that had passed the uncoated porous (8-microm pore size) filters. After staining, filters were examined microscopically for cells on the underside of the membrane. A quantitative protease assay was also performed. Flow cytometric analysis of alpha5 and alpha6 integrin subunits, which are essential for interactions between cells and extracellular matrix, was performed. hCG increased significantly (P<0.01) the in vitro invasion of trophoblastic JEG-3 cells in a dose-dependent manner. Migration was also increased by hCG (P<0.01). However, cell proliferation remained unchanged. The second messenger analogue dibutyryl cAMP (db cAMP) and the cAMP elevating factor (forskolin) mimicked the effects of hCG by stimulating a dose-dependent increase of trophoblastic cell UEG-3) invasion. The collagenolytic activity of trophoblastic cells (EG-3) was increased by hCG stimulation. No changes were shown in the expression of alpha5 and alpha6 integrin subunits on JEG-3 cells. In vitro hCG is a regulatory factor of invasion and migration in trophoblastic JEG-3 cells, whereas proliferation is not influenced. The endogenous production of hCG by the trophoblast in vivo implies an autocrine control of invasion processes by hCG. PMID- 9859864 TI - Expression of endothelial (type III) nitric oxide synthase in cytotrophoblastic cell lines: regulation by hypoxia and inflammatory cytokines. AB - Expression of endothelial nitric oxide synthase (eNOS) has been localized to the villous syncytiotrophoblasts suggesting that NO release from these cells could prevent platelet adhesion and aggregation in the intervillous space. Hypoxia- or inflammation-dependent changes in the release of this vasoactive substance may result in thrombus formation and altered vascular resistance which occur in the placental bed of pre-eclamptic patients. To evaluate the influence of low-oxygen tension and inflammation on eNOS production in the trophoblast steady-state eNOS mRNA and protein levels were investigated in cytotrophoblastic BeWo and Jeg-3 cells cultured at 3.5 per cent oxygen and/or in the presence of the pro inflammatory cytokines IL-1 and TNF-alpha. By RT-PCR and immunocytochemistry we demonstrate that BeWo cells produce eNOS mRNA and protein while eNOS polypeptide was undetectable in JEG-3 cells. In BeWo cells addition of both cytokines decreases eNOS mRNA and protein abundancies within 24 h of incubation while each substance alone had no effect. Compared to controls, the amount of eNOS transcripts was found to be elevated at low-oxygen tension, however, cNOS protein was downregulated after 24 h in the hypoxic environment, as shown by immunocytochemistry and Western blot analysis. Forskolin and methotrexate, which induce biochemical differentiation/ growth arrest in choriocarcinoma cells, stimulate eNOS mRNA and protein synthesis, but cannot overcome the decline of eNOS polypeptide levels during hypoxic incubation. It is speculated that acute hypoxia and inflammation impair eNOS/NO production of the trophoblast in vivo, which might contribute to pathological conditions of gestational diseases. PMID- 9859865 TI - Telomerase activity in human chorionic villi and placenta determined by TRAP and in situ TRAP assay. AB - Telomerase activity (TA) was analysed in human chorionic villi and placenta in normal and abnormal pregnancy using the telomeric repeat amplification protocol (TRAP) and in situ TRAP assay. Twenty chorionic villi specimens and 25 placenta specimens from normal pregnancies were examined as well as placenta specimens from 10 cases of intrauterine growth retardation (IUGR; nine asymmetric and one symmetric). TA was detected in 18 of the 20 (90 per cent) chorionic villi specimens and in 18 of the 25 (72 per cent) placenta specimens from normal pregnancy. However, no or only weak TA was exhibited in the placenta specimens of the nine asymmetric IUGR cases. In situ TRAP assay detected TA in trophoblastic cells from normal pregnancy, but not in trophoblastic cells from cases of asymmetric IUGR. PMID- 9859866 TI - Parallel Doppler assessment of yolk sac and intervillous circulation in normal pregnancy and missed abortion. AB - This study assessed yolk sac morphology and vascularity and intervillous blood flow in normal early pregnancy and missed abortion. Transvaginal colour and pulsed Doppler were used in a prospective analysis of 87 normal pregnancies and 48 missed abortions between 6 and 12 weeks gestation. The Kruskal-Wallis rank test was used to calculate the difference in yolk sac diameter and vascularity visualization rate between gestational weeks. Repeated measures analysis of variance was used for comparison of the intervillous circulation between groups. The growth of the yolk sac was considered statistically significant between gestational weeks 6 and 9, being most prominent between 9 and 10 weeks of gestation. Vascularity of the yolk sac, characterized by low velocity and absence of diastolic flow, was demonstrated in 67 per cent of normal pregnancies. Yolk sac blood flow was detected in 19 per cent of the patients with missed abortion. Doppler analysis of the intervillous circulation demonstrated decreased peak velocity of the continuous flow in patients with missed abortion for gestational weeks 11 and 12. It is concluded that progressive decrease of yolk sac vascularity coincides with visualization of more prominent colour-coded areas within the intervillous space. In patients with missed abortion, such changes do not occur. PMID- 9859867 TI - Interleukin-1beta and bacterial endotoxin change the metabolism of prostaglandins E2 and F2alpha in intact term fetal membranes. AB - There is strong evidence that prostaglandins E2 and F2alpha (PGE2 and PGF2alpha) are involved in the initiation and maintenance of human parturition and that their production can be stimulated by a number of cytokines and in infection induced preterm labour by bacterial endotoxin. This study used an intact fetal membrane disk model to investigate the regulation of PGE2 and PGF2alpha metabolism by interleukin-1 beta (IL-1beta) and bacterial endotoxin [lipopolysaccharide (LPS)]. Fetal membrane explants were incubated with IL-1beta (0.1 or 1.0 ng/ml) or LPS (10 ng/ml) for 24 h. A mixture of 3H-prostaglandin (0.1 microCi) and unlabelled prostaglandin (1 microg) was then added at selected times after the addition of inflammatory mediators. The radiolabelled prostaglandins and their metabolites were then extracted from the culture medium and quantified by high-pressure liquid chromatography. Levels of prostaglandin metabolites were generally decreased following incubation with IL-1beta or LPS, which is consistent with a decrease in the activity of 15-hydroxyprostaglandin dehydrogenase (PGDH). It is concluded that IL-1beta and LPS moderately decrease the metabolism of prostaglandins, which may contribute to increasing the local levels of active prostaglandins induced by these stimuli. PMID- 9859868 TI - Prostaglandin production by guinea-pig placenta and other uterine tissues during mid-pregnancy. AB - Prostaglandin (PG) output from cultured placenta, sub-placenta, endometrium and fetal membranes of guinea-pigs was measured on days 22, 29 and 36 of pregnancy to establish the source of increased PGF2alpha production during mid-pregnancy. PGF2alpha and 6-keto-PGF1alpha were produced in larger quantities than PGE2 by the placenta, sub-placenta and endometrium; 6-keto-PGF1alpha was in the major prostaglandin produced by the fetal membranes. The initial outputs of PGF2alpha, PGE2 and 6-keto-PGF1alpha from the sub-placenta, fetal membranes and endometrium either decreased or remained fairly constant between days 22 and 36. In contrast, the initial outputs of PGF2alpha, PGE2 and 6-keto-PGF1alpha from the placenta increased 14.7-, 2.5- and 2.0-fold, respectively, between days 22 and 36, indicating that the placenta is the tissue responsible for the increase in PGF2alpha output from the mid-pregnant guinea-pig uterus. Aristolochic acid (a phospholipase A2 inhibitor) inhibited prostaglandin output from the endometrium, but had a more variable effect in prostaglandin output from the other tissues. Thimerosal (an arachidonic acid uptake inhibitor) inhibited PGF2alpha and PGE2 outputs from the endometrium, but generally potentiated 6-keto-PGF1alpha output and prostaglandin output from the other tissues. Arachidonic acid release for prostaglandin synthesis in the endometrium, but not the placenta, sub-placental or fetal membranes, is apparently dependent upon a constant level of phospholipase A2 activity. PMID- 9859869 TI - Two-dimensional mapping and microsequencing of lysosomal proteins from human placenta. AB - Lysosomes degrade a wide range of macromolecules to yield monomer products which are exported out of the lysosome by a series of transporters. In addition, lysosomes perform a range of other functions which are cell or tissue specific. In order to gain insight into the tissue specific role of lysosomes, carrier ampholyte two-dimensional electrophoresis (2-DE) was used in combination with N terminal sequencing to identify the major proteins present in both the membrane and luminal space of placental lysosomes. From the 45 N-terminal peptide sequences generated, 14 luminal and five membrane proteins were identified while three other sequences were novel. The sequenced proteins were a mixture of lysosomal and non-lysosomal proteins. The lysosomal proteins consisted of gamma interferon-inducible protein (IP-30), Saposin D, cathepsins B and D, beta hexosaminidase, palmitoyl protein thioesterase, alpha-glucosidase, and LAMP-1. The non-lysosomal proteins were serum albumin, serotransferrin, haemoglobin gamma G chain, alpha-1-antitrypsin, placental lactogen, endoplasmin, peptide binding protein 74, p60 lymphocyte protein, p450 side chain cleavage enzyme and placental alkaline phosphatase. The 2-DE maps obtained in this study are the first to identify the major proteins in both the lumen and membrane of placental lysosomes through sequence analysis, and thus provide the basis upon which to build a complete 2-DE database of the lysosome. Furthermore, the identities of the proteins sequenced from the placental lysosomes suggest a role for lysosomes in the transport of nutrients across the trophoblastic layer. PMID- 9859870 TI - Studies on the localization of 5-hydroxytryptamine and its receptors in human placenta. AB - The localization of 5-hydroxytryptamine (5-HT) and its receptors in human placenta was studied under light and electron microscopy using immunohistochemistry and in situ hybridization. The syncytiotrophoblast, cytotrophoblast, stromal cells and decidual cells in human placentae all appeared to be 5-HT immunoreactive. The 5-HT immunoreactive material was distributed in cytoplasm with negative nuclei. The 5-HT immunoreactive material was also found in capillary endothelium. Trophoblast cells cultured in serum-free medium also showed 5-HT immunoreactivity in the cytoplasm. Fetal white blood cells and both syncytiotrophoblast and cytotrophoblast in human placenta showed 5-HT receptor immunoreactivity and 5-HT1A receptor mRNA hybridized signal was also detected in cytoplasm. The stromal cells and capillary endothelium in placental villi and maternal decidual cells all showed 5-HT receptor immunoreactivity in cytoplasm. The small flattened vesicles and large dense cored vesicles within trophoblast cells showed electron-dense 5-HT receptor immunoreactivity using immunoelectron microscopy. These results suggest that human placenta may not only produce 5-HT but also be a 5-HT target organ, and that 5-HT may not only play roles in placental development and pregnancy maintenance by paracrine and autocrine interactions but may also take part in regulating fetal development. PMID- 9859871 TI - Regulation of the choline transport system in superfused microcarrier cultures of BeWo cells. AB - BeWo choriocarcinoma cells were cultured onto solid microcarrier beads, packed into syringe barrels and superfused. The unidirectional choline uptake across the microvillous membrane of the cells was measured by a rapid single-circulation paired-tracer dilution procedure using methyl[3H]choline with D-[14C]mannitol as the extracellular reference molecule. Choline influx was saturable with a K(t) of 214+/-15 microM and a V(max) of 45.29+/-0.94 nmol/min/mg of cell protein. Uptake of labelled choline was partially inhibited by nicotine, strongly inhibited by hemicolinium-3, and was reduced by about 50 per cent in sodium-free perfusates. A range of agents was added to the stirrer flasks 24 h prior to the experiments to determine if intracellular or extracellular levels of choline or its metabolic product, acetylcholine, regulated choline uptake. Pre-incubation with 2 mM choline reduced the choline maximal uptake by half, while pre-incubation with 100 microM alpha-NETA [2-(alpha-naphthoyl)ethyltrimethyl-ammonium] reduced the influx by 77 per cent. Choline influx was also reduced to about half in the presence of 100 microM vesamicol, bethanecol or neostigmine. It is concluded that BeWo cells possess a choline transporter similar to that described in isolated cytotrophoblasts and syncytiotrophoblast microvillous membrane preparations, and that uptake appeared to be regulated by both intracellular and extracellular concentrations of choline and acetylcholine. Therefore, these cells provide a novel model for studying the role of acetylcholine in human placenta. PMID- 9859872 TI - Hepatocyte growth factor levels during normal and intra-uterine growth-restricted pregnancies. AB - Hepatocyte growth factor (HGF), also known as scatter factor, binds the c-met receptor. It has been shown to be involved in mesenchyme-epithelial interactions. HGF is produced by the villous mesenchyme of the placenta throughout pregnancy and its receptor located on the villous cytotrophoblast cells. In this study the levels of HGF were measured in consecutive samples of plasma taken from pregnant women. Normal pregnancies were compared with intrauterine growth restricted (IUGR) pregnancies (below the third centile). In both groups, the levels of HGF were found to increase significantly as pregnancy progressed and then fall post partum. There was a considerable amount of variation found between individual women but no significant difference (P=0.65) between the normal and IUGR pregnancies. PMID- 9859873 TI - Delayed vasoconstriction of the umbilico-placental circulation by angiotensin in fetal sheep. PMID- 9859874 TI - Are placental-fetal relationships a cause of neurodevelopmental disability? Abstracts and discussions of papers presented at a workshop in Weybridge, UK, on 21-23 November 1996. PMID- 9859875 TI - Dental anthropology of central-southern, Iron Age Italy: the evidence of metric versus nonmetric traits. AB - Discrete and metric dental traits are used to assess biological similarities and differences among 13 bioarchaeological populations located on each side of the Apennine mountains in central-southern Italy and dated to the first millennium BC. An initial hypothesis, that the mountain chain might provide a significant geographical barrier for population movement (resulting in greater biological affinities among those groups on the same side), is not supported. Instead, the samples appear to cluster more on the basis of time than geography. Archaeological evidence, however, supports an association between populations on opposite sides of the mountains and thus is in accord with the dental data. As anticipated, discrete dental traits appear to be more useful than metric dental traits in assessing such population affinities. This research represents a beginning to a better comprehension of the complexity of the biological and cultural dynamics of Italian populations during recent millennia. PMID- 9859876 TI - The posterior border of the sphenoid greater wing and its phylogenetic usefulness in human evolution. AB - The elucidation of patterns of cranial skeletal maturation and growth in fossil hominids is possible not only through dental studies but also by mapping different aspects of ossification in both extant African apes and humans. However, knowledge of normal skeletal development in large samples of extant great apes is flimsy. To remedy this situation, this paper offers an extensive survey and thorough discussion of the ossification of the posterior border of the sphenoid greater wing. Indeed, this area provides much information about basicranial skeletal maturation. We investigate three variants: the absence of the foramen spinosum and the position of both the foramen spinosum and the foramen ovale in relation to the sphenosquamosal suture. Providing original data about humans and 1,425 extant great ape skulls and using a sample of 64 fossil hominids, this study aimed to test whether different ossification patterns occurred during the course of human evolution. The incidence of three derived morphologies located on the posterior border of the sphenoid greater wing increases during human evolution at different geological periods. The evolutionary polarity of these three derived morphologies is assessed by outgroup comparison and ontogenetic methods. During human evolution, there is a clear trend for the foramen spinosum to be present and wholly located on the posterior area of the sphenoid greater wing. Moreover, in all the great ape species and in Australopithecus afarensis, the sphenosquamosal suture may split the foramen ovale. Inversely, the foramen ovale always lies wholly within the sphenoid greater wing in Australopithecus africanus, robust australopithecines, early Homo, H. erectus (and/or H. ergaster), and Homo sapiens. From ontogenetic studies in humans, we conclude that, during human evolution, the ossification of the posterior area of the sphenoid greater wing progressively surrounded the middle meningeal artery (passing through the foramen spinosum) and the small meningeal artery (passing through the foramen ovale). PMID- 9859877 TI - Correlated response, competition, and female canine size in primates. AB - Recently, comparative analyses of female canine tooth size in primates have yielded two hypotheses to explain interspecific variation in female relative canine size. Greenfield ([1992] Int. J. Primatol. 13:631-657; [1992] Yrbk. Phys. Anthropol. 35:153-184; [1996] J. Hum. Evol. 31:1-19) suggested that covariation in male and female canine size across species indicates that female canine size reflects correlated response (in which the expression of a trait in one sex causes the expression of the same trait in the other sex). Plavcan et al. ([1995] J. Hum. Evol. 28:245-276) noted that female canine size in primates is associated with variation in categorical estimates of the intensity of female-female agonistic competition, suggesting that selection favors large female canine size in many species. While it may seem that the two models are in conflict, they are not. To simultaneously evaluate these two models, this analysis examines the joint relations between male canine size, female canine size, and estimates of female-female competition in a sample of 108 primate species. Overall, female canine size is correlated with variation in male canine size. Controlling for variation in male canine size, female canine size is also correlated with estimates of the intensity of female-female agonistic competition. The relation between these variables differs strongly between anthropoid and strepsirhine primates. In anthropoids, the data suggest that selection for the development of large canines in females is not constrained by any affect of correlated response. In strepsirhines, the evidence suggests that sexual selection may affect male canine size but that correlated response affects female canine size, resulting in monomorphism for most species. These observations help reconcile the observations of Greenfield ([1992] Int. J. Primatol. 13:631-657; [1996] J. Hum. Evol. 31:1-19) and Plavcan et al. ([1995] J. Hum. Evol. 28:245-276) and provide a more precise model for understanding interspecific variation in female canine size and hence canine dimorphism. PMID- 9859879 TI - Proceedings of the Sixty-Seventh Meeting of the American Association of Physical Anthropologists. Salt Lake Hilton. Salt Lake City, Utah. April 1-April 5, 1998. PMID- 9859878 TI - Brief communication: twentieth-century replication of an Egyptian mummy: implications for paleopathology. AB - Replication in a modern human cadaver of ancient Egyptian mummification focused on tools used by ancient Egyptian embalmers, the use of natron (a mixture of sodium carbonate, bicarbonate, and chloride) in the preparation of the mummy, surgical procedures in the removal of the viscera and brain, and histologic examination of the viscera. The first three areas have been reported separately (Brier and Wade [1997] ZAS 124:89-100). In this paper, we demonstrate a degree of histologic preservation comparable to that seen in Egyptian mummies, indicating the effectiveness of ancient mummification and that the histologic appearance of such mummies is little altered by the passage of millennia. PMID- 9859880 TI - A tutorial on conducting meta-analyses of clinical outcome research. AB - Throughout the educational, medical, psychological, and social sciences, meta analysis is the present-day, broadly accepted means for combining many quasi experiments in a synthesis for the purpose of establishing the weight of scientific evidence bearing on a certain research question. Meta-analysis thereby is the preferred method for determining the preponderance of evidence in clinical outcome research relating to questions of treatment efficacy and treatment effectiveness. Relatively few meta-analyses appear in the literature of the communication disorder sciences. The purpose of this tutorial is to enhance the familiarity and accessibility of this technology in the domains of audiology and speech-language pathology. The results of the accompanying example constitute a preliminary meta-analysis of patient-perceived treatment effectiveness. The substance of the tutorial, however, transcends disciplinary interests regarding types of communication disorder. PMID- 9859881 TI - Two- to six-year controlled-trial stuttering outcomes for children and adolescents. AB - This research is a long-term follow-up of a previously published, controlled trial on the effectiveness of 3 stuttering treatments (intensive smooth speech, parent-home smooth speech, and intensive electromyography feedback) for children and adolescents, aged 11 to 18 years, who stutter. The previous controlled trial showed all 3 treatments to be effective compared to nontreatment after 12 months. This paper reports on the treatment effectiveness after an average of 4 years post-treatment. Results demonstrate that treatment gains were maintained in the long term, with rates of stuttering similar to the 1-year postoutcomes. There were no significant differences among the 3 treatments in long-term effectiveness. This controlled study substantiates the claim that the treatments investigated will more than likely have substantial long-term benefits for the fluency and personality of children and adolescents who stutter. PMID- 9859882 TI - Enhancement of electrolaryngeal speech by adaptive filtering. AB - Artificial larynges provide a means of verbal communication for people who have either lost or are otherwise unable to use their larynges. Although they enable adequate communication, the resulting speech has an unnatural quality and is significantly less intelligible than normal speech. One of the major problems with the widely used Transcutaneous Artificial Larynx (TAL) is the presence of a steady background noise caused by the leakage of acoustic energy from the TAL, its interface with the neck, and the surrounding neck tissue. The severity of the problem varies from speaker to speaker, partly depending upon the characteristics of the individual's neck tissue. The present study tests the hypothesis that TAL speech is enhanced in quality (as assessed through listener preference judgments) and intelligibility by removal of the inherent, directly radiated background signal. In particular, the focus is on the improvement of speech over the telephone or through some other electronic communication medium. A novel adaptive filtering architecture was designed and implemented to remove the background noise. Perceptual tests were conducted to assess speech, from two individuals with a laryngectomy and two normal speakers using the Servox TAL, before and after processing by the adaptive filter. A spectral analysis of the adaptively filtered TAL speech revealed a significant reduction in the amount of background source radiation yet preserved the acoustic characteristics of the vocal output. Results from the perceptual tests indicate a clear preference for the processed speech. In general, there was no significant improvement or degradation in intelligibility. However, the processing did improve the intelligibility of word initial non-nasal consonants. PMID- 9859883 TI - Adaptation of stuttering frequency during repeated readings: associated changes in acoustic parameters of perceptually fluent speech. AB - This study is part of a series investigating the hypothesis that stuttering adaptation is a result of motor learning. Previous investigations indicate that nonspeech motor learning typically is associated with an increase in speed of performance. Previous investigations of stuttering, on the other hand, indicate that improvements in fluency during most fluency-enhancing conditions or after stuttering treatment tend to be associated with decreased speech rate, increased duration of specific acoustic segments, and decreased vowel duration variability. The present acoustic findings, obtained from 8 individuals who stutter, reveal that speech adjustments occurring during adaptation differ from those reported for other fluency-enhancing conditions or stuttering treatment. Instead, the observed changes are consistent with those occurring during skill improvements for nonspeech motor tasks and, thus, with a motor learning hypothesis of stuttering adaptation. During the last of 6 repeated readings, a statistically significant increase in articulation rate was observed, together with a decrease in word duration, vowel duration, and consonant-vowel (CV) transition extent. Other adjustments showing relatively consistent trends across individual subjects included decreased CV transition rate and duration, and increased variability of both CV transition extent and vowel duration. PMID- 9859884 TI - Dysarthric sentence intelligibility: contribution of iconic gestures and message predictiveness. AB - This study examined changes in the sentence intelligibility scores of speakers with dysarthria in association with different signal-independent factors (contextual influences). This investigation focused on the presence or absence of iconic gestures while speaking sentences with low or high semantic predictiveness. The speakers were 4 individuals with dysarthria, who varied from one another in terms of their level of speech intelligibility impairment, gestural abilities, and overall level of motor functioning. Ninety-six inexperienced listeners (24 assigned to each speaker) orthographically transcribed 16 test sentences presented in an audio + video or audio-only format. The sentences had either low or high semantic predictiveness and were spoken by each speaker with and without the corresponding gestures. The effects of signal independent factors (presence or absence of iconic gestures, low or high semantic predictiveness, and audio + video or audio-only presentation formats) were analyzed for individual speakers. Not all signal-independent information benefited speakers similarly. Results indicated that use of gestures and high semantic predictiveness improved sentence intelligibility for 2 speakers. The other 2 speakers benefited from high predictive messages. The audio + video presentation mode enhanced listener understanding for all speakers, although there were interactions related to specific speaking situations. Overall, the contributions of relevant signal-independent information were greater for the speakers with more severely impaired intelligibility. The results are discussed in terms of understanding the contribution of signal-independent factors to the communicative process. PMID- 9859885 TI - FO processing and the separation of competing speech signals by listeners with normal hearing and with hearing loss. AB - Normal-hearing and hearing-impaired listeners were tested to determine F0 difference limens for synthetic tokens of 5 steady-state vowels. The same stimuli were then used in a concurrent-vowel labeling task with the F0 difference between concurrent vowels ranging between 0 and 4 semitones. Finally, speech recognition was tested for synthetic sentences in the presence of a competing synthetic voice with the same, a higher, or a lower F0. Normal-hearing listeners and hearing impaired listeners with small F0-discrimination (deltaF0) thresholds showed improvements in vowel labeling when there were differences in F0 between vowels on the concurrent-vowel task. Impaired listeners with high deltaF0 thresholds did not benefit from F0 differences between vowels. At the group level, normal hearing listeners benefited more than hearing-impaired listeners from F0 differences between competing signals on both the concurrent-vowel and sentence tasks. However, for individual listeners, deltaF0 thresholds and improvements in concurrent-vowel labeling based on F0 differences were only weakly associated with F0-based improvements in performance on the sentence task. For both the concurrent-vowel and sentence tasks, there was evidence that the ability to benefit from F0 differences between competing signals decreases with age. PMID- 9859886 TI - The dependence of the distortion product 2f1-f2 on primary levels in non-impaired human ears. AB - The optimal intensity relation between the two primaries used to generate Distortion Product Otoacoustic Emissions (DPOAEs) in unimpaired human ears, over a clinically relevant intensity range, was evaluated using a commercially available clinical device. The ILO92 was used to determine the level of the DPOAE at 2f1-f2 for 16 combinations of primary levels in the range of 40 to 80 dB SPL from 40 unimpaired, young adult ears. Data were collected between 1 and 6 kHz at 1, 1.5, 2, 3, 4, 5, and 6 kHz. The commonly used procedure of dropping data points less than 3 dB above the noise floor was compared to a power subtraction procedure. A multivariate ANOVA was performed to determine main effects of gender, ear, stimulus levels, frequencies, and interactions between stimulus levels and frequencies. An overall increase of DPOAE amplitude with increase in primary level was observed, along with a decrease of the optimal difference in primary levels as L2 was increased. Although the power subtraction and 3-dB drop paradigms yielded similar results at high stimulus levels, the power subtraction paradigm provided a more realistic indicator of DPOAE level when low level primaries were used. Possible mechanisms responsible for the level dependence of the optimal relationship between primaries and implications for clinical choice of primary levels are discussed. PMID- 9859887 TI - Transient-evoked otoacoustic emissions as a measure of noise-induced threshold shift. AB - Otoacoustic emissions and behavioral hearing thresholds were measured before and after exposure to a 10-min, 105-dB SPL, half-octave band of noise centered at 1.414 kHz. Along a single recovery function, transient-evoked otoacoustic emission (TEOAE) measurements made with 74-dB pSPL nonlinear click ensembles were alternated with a Bekesy threshold-tracking procedure. Each of the 14 participants with normal hearing underwent 2 hour-long temporary-threshold shift (TTS) sessions as well as 2 pretest sessions and a posttest session. The Bekesy test frequency was fixed at 2.0 kHz, whereas emissions were analyzed in half octave bandwidths with center frequencies ranging from 0.707 to 5.656 kHz. Results showed that (a) the maximum temporary emission shifts (TES) were half to 1 octave above the exposure frequency; (b) the 4.7-dB average temporary emission shift magnitude at approximately 2 min postexposure was less than half of the 11.7-dB average TTS; (c) the average recovery times for emissions and hearing thresholds were similar (188 vs. 186 min); and (d) the average TTS magnitude along the recovery function was predictable from TES magnitude. It is concluded that both TEOAEs and Bekesy thresholds reveal the same aspects of postexposure inner-ear changes. PMID- 9859888 TI - HINT list equivalency using older listeners. AB - HINT list equivalency was examined using 24 listeners between 60 and 70 years old who had sensorineural hearing impairment. A Greco-Latin square design was used to ensure that each list was presented an equal number of times per condition. Four conditions were tested: (1) speech in quiet, (2) speech in 65 dBA noise with noise at 0 degrees azimuth, (3) speech in 65 dBA noise with noise at 90 degrees azimuth, and (4) speech in 65 dBA noise with noise at 270 degrees azimuth. Speech materials were always presented at 0 degrees azimuth. Overall mean scores ranged from 29.9 dBA for the quiet condition to 63.4 dBA for the noise at 0 degrees azimuth condition. A significant difference was found between Lists 13 and 16 only. This was attributed to audibility differences among the listeners. Therefore, the 25 HINT lists should be considered equivalent for older populations with similar hearing impairment. The HINT lists can be used for relative measures, such as comparison of aided versus unaided sentence SRTs or comparison of 2 different hearing aids. PMID- 9859889 TI - Analysis of observational data in speech and language research using generalizability theory. AB - Most research in speech-language pathology relies on observational data collected by human observers or judges. The reliability and generalizability of such measurements are always important considerations. This article reviews classical methods of estimating reliability and proposes that a more powerful approach capable of estimating the dependability of behavioral measurements is available. This approach, based on generalizability theory, provides a practical framework for estimating multiple sources of measurement error in the collection of observational data. Concepts central to generalizability theory are discussed, and a hypothetical data set illustrates the usefulness of generalizability measurements in speech and language research. PMID- 9859890 TI - Understanding referential expressions in context: use of common ground by children and adolescents with mental retardation. AB - Listeners interpret utterances against the common ground, or network of presuppositions shared with the speaker. The first purpose of the study was to determine whether individuals with mental retardation use the major sources of common ground (i.e., physical copresence, linguistic copresence, and community membership) to resolve referential ambiguity. The second purpose was to determine whether they seek confirmation of their referent choices in accordance with the certainty of interpretation afforded by the common ground. The third purpose was to determine whether they signal noncomprehension when faced with ambiguity and common ground that is not informative. The final purpose was to evaluate the relationship between within-group variability in common ground use and measures of nonverbal cognition, receptive and expressive language, and social cognition. Participants were school-age individuals with mental retardation and typically developing children matched to them on nonverbal MA. Common ground use was examined in a role-playing task in which the participant responded to ambiguous utterances. Common ground was manipulated within participants. We determined whether referent selections were appropriate for the common ground, whether they were accompanied by confirmation requests, and whether noncomprehension was signaled. Both groups used all sources of common ground to resolve referential ambiguity at better than chance levels but were less successful in using community membership. Both groups also requested confirmation of their referent choices most often when the common ground was based on community membership. Both groups signaled noncomprehension when the common ground was not informative. Different aspects of common ground use were related to different predictors for the group with mental retardation. PMID- 9859891 TI - Grammatical morphology and the role of weak syllables in the speech of Italian speaking children with specific language impairment. AB - Italian-speaking children with specific language impairment (SLI) were compared to a group of younger control children in their use of auxiliary verbs, pronominal clitics, infinitives, present tense verb inflections, and articles. Differences favoring the control children were found for those morphemes that required the production of nonfinal weak syllables. On other grammatical morphemes, the two groups did not differ. A relationship was seen between the use of morphemes requiring nonfinal weak syllables and the use of nonfinal weak syllables that had no morpheme status. The findings are considered from the perspective of both prosodic production limitations and limitations in input processing. PMID- 9859892 TI - The effect of presentation rate on serial memory in young children with specific language impairment. AB - The study examined the serial memory ability of a group of preschool children with specific language impairment (SLI) who were compared to age and language control groups. The children were asked to recognize serial patterns under short and long presentation durations. The subjects were presented with images of common objects (that appeared to be easily recoded into a phonological form) and iconic images of scribble drawings and unfamiliar faces (that did not appear to invite recoding). Under long presentation conditions, the performance of children with SLI resembled that of their age-matched peers on all 3 types of tasks. However, under short presentation conditions, children with SLI performed worse than their age-matched peers on all 3 tasks (and similarly to their language matched peers). The performance of the children with SLI declined dramatically in all conditions when the items were presented for a brief period. If the serial memory deficits of young children with SLI were specific to phonological processing, their performance on recognizing the pattern of common objects should have been impaired, but not their performance with other visual tasks that are less likely to be recoded. Instead, serial memory in children with SLI was affected by presentation duration across tasks. The findings suggest that recognizing serial patterns is dependent, in part, on the speed of processing serial information. The findings are discussed in relation to models of limited capacity processing. PMID- 9859893 TI - Early verb-related vulnerability among children with specific language impairment. AB - The purpose of this study was to characterize the nature of early grammatical development among very young children with specific language impairment (SLI). Grammatical development was examined for two subtypes: (a) children with expressive language impairments only (SLI-E) and (b) children with both receptive and expressive language impairments (SLI-RE). In particular, characteristics of noun-phrase (NP) and verb-phrase (VP) elaboration were examined longitudinally to determine whether structures associated with NP and VP emerged together following a typical developmental progression. Group analyses did not reveal any differences between the subtypes on the Index of Productive Syntax (IPSyn; Scarborough, 1990). However, specific weakness in VP elaboration was revealed on the IPSyn as well as in more extensive productivity analyses. The contribution of these findings to a developmentally sensitive grammatical description of SLI for very young children is discussed. PMID- 9859894 TI - Identifiers of predominantly Spanish-speaking children with language impairment. AB - The purpose of this study was to identify a set of measures that would discriminate 31 predominantly Spanish-speaking children with normal language (NL children) from 31 children with language impairment (LI children). The LI children were identified as such by experienced, bilingual (Spanish/English), ASHA-certified, speech-language pathologists who were currently seeing the children in their caseloads. Children ranged in age from 5 to 7 years and were matched for age, gender, and school. Additionally, nonverbal cognitive measures assured that they did not differ significantly intellectually. Measures of vocabulary, novel bound-morpheme learning skills, and language form were randomly administered to all children. Further, parents responded to questions about their perceptions of their children's speech and language skills and family history of speech and language problems. A stepwise discriminant analysis indicated that 4 measures discriminated the groups of children with a sensitivity of 91.3% and a specificity of 100% (p < .0001): parental report of the child's speech and language skills, number of errors per T-unit, mean length per T-unit, and family history of speech and language problems. A second discriminant analysis indicated that the sensitivity and specificity could be maintained when only the first 2 measures were included. Confirmatory discriminant analyses of the 2- and 4 measure models indicated that the discriminant accuracy was stable on an independent sample. PMID- 9859895 TI - Tense over time: the longitudinal course of tense acquisition in children with specific language impairment. AB - Tense marking in English is relatively late appearing and is especially late for children with Specific Language Impairment (SLI). Little is known about the full course of acquisition for this set of morphemes. Because tense marking is a fundamental property of clause construction, it is central to current theories of morphosyntax and language acquisition. A longitudinal study is reported that encompasses the years of 2;6-8;9 years for typically developing children (N = 43) and 4;6-8;8 years for children with SLI (N = 21). The findings show that a diverse set of morphemes share the property of tense marking; that this set is not mastered until age 4 years in typically developing children and after 7 years for children with SLI; that acquisition shows linear and nonlinear components for both groups, in a typical S-shaped curve; and that nonsyntactic measures are not predictors of growth (including nonverbal intelligence, vocabulary size, and mother's education), whereas initial MLU does predict rate of acquisition. The findings are consistent with a model of Optional Infinitives (OI) for typically developing children (cf. Wexler, 1994, 1996) and Extended Optional Infinitives (EOI) for children with SLI. This model hypothesizes incomplete specification of features of tense that are represented in the grammar. PMID- 9859896 TI - Real-time inflectional processing by children with specific language impairment: effects of phonetic substance. AB - In this study, we examined the processing of low-phonetic-substance inflections (e.g., third-person-singular -s, past-tense -ed) versus a higher-phonetic substance inflection (e.g., present-progressive -ing) by children with specific language impairment (SLI) in two types of receptive tasks. Twenty-one children with SLI (Age: 8 years;6 months), 21 chronological age matched (CA; Age: 8;7), and 21 receptive syntax matched (RS; Age: 6;8) children participated in a word recognition reaction time (RT) task and an off-line task requiring grammaticality judgments. On the RT task, the children with SLI demonstrated RT sensitivity only to the presence of a higher-phonetic-substance inflection, unlike the CA and RS controls who displayed sensitivity to both higher-substance and low-substance inflections. On the grammaticality judgment task, the children with SLI performed more poorly than the CA controls only on sentences missing obligatory low substance inflections (e.g., "Carl already jump over the fence"). The findings are discussed within the framework of the surface account, which predicts that children with SLI should have greater difficulty processing and making grammatical judgments about low-substance inflections compared to higher substance inflections. PMID- 9859897 TI - The impact of emphatic stress on novel word learning by children with specific language impairment. AB - This investigation examined the influence of emphatic stress on children's novel word learning. Forty school-age children participated in this study, including 20 children with specific language impairment (SLI) and 20 children with normal language (NL) development. Results indicated that there were no significant stress effects for comprehension or recognition of novel words (for which all children demonstrated relatively high levels of performance); however, children in both groups exhibited significantly better production of words that had been presented with emphatic stress than with neutral stress. These findings are discussed within a limited capacity framework of language processing. PMID- 9859898 TI - Differential gene expression at gestational days 14 and 16 in normal and nitrogen induced hypoplastic murine fetal lungs with coexistent diaphragmatic hernia. AB - The purpose of this study was to identify differentially expressed genes in normal and nitrofen-induced hypoplastic lungs in fetal mice. Such genes may play a role in the regulation of lung development. CD-1 pregnant dams were gavaged with 25 mg of nitrofen on gestational day (Gd) 8 to induce pulmonary hypoplasia and diaphragmatic hernia (DH). Normal and nitrofen-treated fetuses were removed on Gd 14 and Gd 16. Lungs were examined in all nitrofen-exposed fetuses and only those that had developed severely hypoplastic lungs with coexistent diaphragmatic hernia were taken for molecular analyses. RNA was extracted from normal and nitrofen-treated lungs, reverse transcribed, and PCR-amplified using 48 combinations of anchor and arbitrary primers for each condition. The resulting cDNAs from normal and hypoplastic lungs were run on 6% polyacrylamide differential display gels. In Gd 14 lungs, we observed 10 differentially expressed cDNA bands, of which 6 were identified to be inhibited and 4 were reduced in the hypoplastic lungs compared to normal fetal lungs. From the Gd 16 lungs, a total of 29 differentially expressed cDNA bands were found, of which 11 were reduced, 4 were inhibited, 11 were enhanced, and 3 were induced in the hypoplastic compared to the normal lungs. All 39 differentially expressed cDNAs were cloned, sequenced, and identified through BLAST searches. Among the sequences that were identified, results were as follows: 1) Hypoplastic Gd 14 lungs had two unknown cDNA sequences with reduced/inhibited expressions, whereas one was a known sequence having 77% similarity with a promoter region regulating various cytokines such as IL-1, IL-2, and IL-11. The expression of this sequence was inhibited in the hypoplastic lungs. This sequence also had similarity to lipid-binding proteins. 2) On Gd 16, hypoplastic lungs had one cDNA sequence with reduced expression which had 82% similarity with thyroid hormone receptor gene exon 1 and two other cDNA sequences with enhanced expressions. One of these enhanced cDNA sequences in hypoplastic lungs had 98% similarity with the fibroblast growth factor receptor-3 gene, and the other was an unknown sequence. Northern blot hybridizations were performed to confirm the differential expression of the two sequences of interest, which were identified as thyroid hormone receptor and fibroblast growth factor (FGF) receptor-3. Overall, out of a total of 39 RT-PCR products (i.e., cDNAs), the abundance of which was altered by nitrofen, 6 were found to be homologous to sequences in Gen Bank through BLAST searches. These 6 sequences became the products of interest, and 3 of these 6 products were similar to previously identified genes. Our results may shed some light on regulatory aspects of lung development and open avenues for treatment of hypoplastic lungs and other respiratory problems in human neonates. PMID- 9859899 TI - Ventilatory failure during resistive loaded breathing in the newborn primate. AB - Previous investigations have shown that ventilatory failure during severe inspiratory resistive loading (IRL) in the 21-day-old infant primate occurs secondary to a decrease in respiratory frequency, that is, central failure. To examine the response of the more immature newborn to IRL, minute ventilation (V'E), arterial blood gases and pH, minute diaphragmatic electromyogram (EMG) activity, peak inspiratory airway pressure, and the centroid frequency (Fc) of the diaphragmatic EMG power spectrum were measured in four unanesthetized tracheotomized 2-day-old monkeys during various levels of IRL, until either 1) ventilatory failure occurred (ventilatory failure run) or 2) normocapnia was sustained for 1 hr (successful trial). During successful trials, minute ventilation, breathing frequency, tidal volume, Fc, and PaCO2 were sustained at baseline levels and an increase in minute EMG activity and peak inspiratory airway pressure were observed. In contrast, during ventilatory failure runs, minute ventilation and tidal volume fell and PaCO2 rose compared to their respective baseline values. Respiratory frequency did not change. The decline in tidal volume occurred despite significant increases in minute diaphragmatic EMG activity and peak inspiratory airway pressure. No shifts in Fc were noted, suggesting that peripheral diaphragmatic fatigue did not occur. We conclude that ventilatory failure during IRL in the 2-day-old monkey is due to the animal's inability to defend tidal volume as opposed to central failure. PMID- 9859900 TI - Dynamics of spontaneous breathing during patient-triggered partial liquid ventilation. AB - This study evaluates different ventilator strategies during gas (GV) and partial liquid ventilation (PLV) in spontaneously breathing animals. We hypothesized that during PLV, spontaneously breathing animals would self-regulate respiratory parameters by increasing respiratory rate (RR) and minute ventilation (V'E) when compared to animals mechanically ventilated with gas, and further that full synchronization of each animal's effort to the ventilator cycle would decrease RR at stable tidal volumes (V(T)). We studied 12 newborn piglets (1.54 +/- 0.24 kg) undergoing GV and PLV in 3 different modes: intermittent mandatory ventilation (IMV), synchronized IMV (SIMV), and assist control ventilation (AC). Modes occurred sequentially in random order during GV first, with the same order then repeated during PLV. Animals initially received continuous positive airway pressure (CPAP) and returned to CPAP during PLV at the end of the experiment. Pressure-limited, volume-targeted ventilation was used with a tidal volume goal of 13 cc/kg. Rate was set at 10/min during IMV and SIMV, with a back-up rate of 10/min during AC. RR, V'E, mechanical (V(T)) and spontaneous tidal volumes (sV(T)) were measured breath-to-breath using a computer-assisted lung mechanics analyzer; mean values were determined over 30-min periods. Data analysis used paired t-tests with Bonferroni correction as needed (P < 0.05). Blood gases were stable in all modes during GV and PLV. RR (min(-1)) and V'E (L x min(-1)/kg) increased in all modes from GV to PLV (RR: CPAP 71 vs. 128; IMV 69 vs. 112; SIMV 65 vs. 107; AC 33 vs. 47. V'E: CPAP 0.47 vs. 0.72; IMV 0.46 vs. 0.61; SIMV 0.45 vs. 0.61; AC 0.38 vs. 0.53; P < 0.05). Intermode comparisons during PLV showed a lower RR with AC (P < 0.02), and a higher V'E with CPAP (P < 0.05). V(T) and dynamic respiratory system compliance decreased from GV to PLV (V(T) P < 0.05; C(rs,dyn) P < 0.01); sV(T) remained unchanged. V(T) and sV(T) did not differ in intermode comparisons. We conclude that during PLV, spontaneously breathing piglets with normal lungs maintain physiologic blood gases by increasing V'E through increased RR. AC produced the most efficient respiratory pattern during PLV, with increased V'E achieved by a modest increase in RR. PMID- 9859901 TI - Urinary bombesin-like peptide levels in infants and children with bronchopulmonary dysplasia and cystic fibrosis. AB - Compared to normal infants and children, there are increased numbers of neuroendocrine cells with bombesin-like peptide (BLP) immunostaining in the lungs of infants and children with bronchopulmonary dysplasia (BPD) and cystic fibrosis (CF). However, there are no data documenting levels of urinary BLP in normal infants and children, or in children with lung disease. We therefore determined the normal developmental pattern for urinary BLP excretion in healthy infants and children, and in infants and children with BPD and CF, and correlated these findings with the subjects' clinical course. We measured urinary BLP levels in 110 subjects: 54 controls, 33 with BPD, and 23 with CF. An age-dependent decline in urinary bombesin levels was evident in the control and BPD subjects, but not in those with CF. There were no statistically significant differences in BLP levels between normal infants and those with BPD. Mean BLP levels were higher in the more immature preterm infants with BPD who required increased ventilatory support. The highest mean BLP levels were documented in BPD infants under age 3 months (882 fmol/mg creatinine), in controls between 3 and 12 months of age (625 fmol/mg creatinine), and in 12-60-month-old CF subjects (486 fmol/mg creatinine). Thus there is an age-dependent decline in BLP levels in controls and BPD, but not in CF. We speculate that the elevated urinary BLP levels in infants and children with BPD and CF may reflect increased pulmonary neuroendocrine cell activity in these conditions, due to the epithelial regenerative response to airway damage and repair. PMID- 9859902 TI - Chronic interstitial lung disease in children: response to high-dose intravenous methylprednisolone pulses. AB - The prognosis for children with chronic interstitial lung disease is poor and the mortality rate is high, especially in infants. This explains the many therapeutical protocols which have been proposed and investigated by several authors. In the present work, we evaluated the response of three infants with idiopathic pulmonary fibrosis to high-dose intravenous prednisolone pulses. The patients were referred to the department at the age of 4, 17, and 3 months, respectively. The diagnosis was confirmed by open lung biopsy and intravenous pulse methyl prednisolone therapy was started with the following protocol: 300 mg/m2 methylprednisolone daily for 3 days, repeated every 4 to 6 weeks. Because of the extreme severity of the respiratory distress at the time of diagnosis, the intravenous pulse treatments were initially complemented by oral prednisone. Clinical improvement was noticed within 6 months with progressive correction of hypoxemia. After follow-up for 3.5 to 4 years, with a total number of pulses of 37, 26, and 32, respectively, the children are symptom-free and do not require oxygen supplementation. During this period, no side effects and no adrenal insufficiency could be documented. Based on current knowledge of steroid action, it can be speculated that the response to intermittent high-dose intravenous methylprednisolone may explain the ability of this mode of hormone administration to maintain an adequate level of glucocorticoid receptor expression. More information and trials through multicenter collaborations are needed to assess therapeutical protocols of repeated high-dose intravenous steroid treatment. PMID- 9859903 TI - Effects of inhaled corticosteroids and inhaled cromolyn sodium on urinary growth hormone excretion in asthmatic children. AB - Over the past few years there has been an increasing awareness that asthma is a chronic inflammatory airways disease. The current therapeutic strategies for treating asthma focus on suppressing the inflammatory process by using cromones or inhaled corticosteroids (ICS). The beneficial effects of ICS in asthma are now well known, but its detrimental effect on linear growth remains a controversial issue. The aim of this open label, nonrandomized, cross-sectional, one-time study was to determine the influence of these drugs on urinary growth hormone (U-GH) levels in prepubertal asthmatic children. U-GH levels were measured in 47 prepubertal asthmatic children who had been treated for at least 6 months with either ICS (beclomethasone or budesonide at a mean daily dose of 360 microg) or with 80 mg daily dose of cromolyn sodium (CrS). There were also nine healthy children who served as a control. These three groups of children were matched for age and gender ratio. The mean level of U-GH in the CrS-treated group was 2.94 +/ 0.96 ng/night; this was significantly higher compared to the mean level of the ICS-treated group (1.99 +/- 0.83 ng/night; P < 0.001) and to the mean level of the control group (1.98 +/- 0.39 ng/night; P < 0.006). There was no significant difference between the mean level of U-GH in the group treated by ICS and the controls (P < 0.9). These results show that the mean levels of U-GH secretion of the children who were treated by CrS for 6 months was significantly increased, compared to the mean U-GH level of the ICS-treated group and the controls. The mean U-GH levels in the last two groups showed no statistically significant difference. PMID- 9859904 TI - Efficacy of bronchodilators administered by nebulizers versus spacer devices in infants with acute wheezing. AB - The aim of this study was to compare the response of infants with acute wheezing to treatments with inhaled terbutaline when administered by nebulizer or by metered-dose inhaler and spacer device (MDI-spacer). Thirty-four infants between the ages of 1 and 24 months who were seen in our emergency department for acute wheezing were studied in a double-blind, randomized trial. The participants received two treatments of terbutaline at 20-min intervals, either by a nebulizer (2 mg/dose in 2.8 mL of 0.9% saline solution) or by an MDI-spacer device (0.5 mg/dose). The outcome measure was a clinical score, based on respiratory rate, degree of wheezing, retractions, degree of cyanosis, color, and pulse oximetry data measured before treatment, 20 min after the first treatment, and again 20 min after the second treatment. There was no difference in the rate of improvement in the clinical score between infants who received terbutaline by nebulizer and those who received it by MDI-spacer. We conclude that MDI-spacers and nebulizers are equally effective means of delivering beta-2 agonists to infants and small children with acute wheezing. PMID- 9859905 TI - Efficacy of nasal intermittent positive pressure ventilation in treating apnea of prematurity. AB - The efficacy of nasal intermittent positive pressure ventilation (NIPPV) in treating apnea of prematurity was evaluated. Apneic preterm infants were randomly assigned to receive either NIPPV or continuous positive airway pressure (NCPAP) for 4 hr when they failed to respond to conservative therapy. The amount of reduction in apneic spells and bradycardia in the two groups after treatment was compared. Thirty-four infants (18 with NIPPV, 16 with NCPAP) were enrolled. Their birth weights ranged from 590-1,880 g (mean, 1,021 g) and gestational ages from 25-32 weeks (mean, 27.6 weeks). The baseline characteristics were comparable in the two groups. Frequency of apnea and bradycardia was reduced during both forms of treatments. However, the infants receiving NIPPV had a greater reduction of apneic spells (P = 0.02) and a tendency to greater decrease in bradycardia (P = 0.09) than those receiving NCPAP. We conclude that NIPPV is more effective than NCPAP in reducing apnea in preterm infants. NIPPV may reduce bradycardia; however, this needs to be validated by a larger number of observations. PMID- 9859906 TI - Acoustic method to quantitatively assess the position and patency of infant endotracheal tubes: preliminary results in rabbits. AB - In this preliminary laboratory study, an acoustical method was evaluated to quantitatively assess the position and patency of an infant-size endotracheal tube (ETT) by in vivo and in vitro measurements. The method consists of emitting an audible sound pulse into the ETT and the airways, and deriving position and patency information from the timing and characteristics of the returning echoes. The method's capacity to measure ETT changes of position in the tracheae of five anesthetized New Zealand white rabbits (weight, 4.3-4.9 kg; age, 1.5-3 years) was found to be accurate to 0.7 +/- 3.6 mm (mean +/- 95% CI) over a distance of 5 cm. The method was also shown to reliably differentiate between tracheal, bronchial, and esophageal intubations by means of an acoustically inferred diameter of the passageway just beyond the ETT tip. To assess the accuracy of estimating lumen obstruction, in vitro acoustical measurements were performed in different size ETTs (2.5, 3.0, 3.5, and 4.0 mm inner diameter), with obstructions ranging from 5 100% reduction in cross-sectional area. The system identified the sizes of these obstructions to within +/-7%. This technology has the potential for continuous, computer-based monitoring of breathing-tube function through instantaneous detection of ETT malposition or obstruction before it leads to a serious medical condition. PMID- 9859907 TI - A steroid-induced acute psychosis in a child with asthma. PMID- 9859910 TI - Thalidomide and its impact in dermatology. AB - Thalidomide, originally marketed as a sedative, was introduced in West Germany in 1956 and in numerous other countries soon thereafter. In part because it did not impair coordination or respiratory function, the drug rapidly became extremely popular. By 1961, however, there were mounting reports of phocomelia and other severe congenital abnormalities associated with maternal use of thalidomide, and the drug was withdrawn from the market and its availability highly restricted. A few years later, thalidomide would find use in dermatology after it was reported that leprosy patients with erythema nodosum leprosum (ENL) experienced rapid and dramatic improvement after taking the drug as a sedative. Additional data quickly confirmed thalidomide's efficacy in ENL, and today it is the drug of choice in the condition. In subsequent decades, the drug has been successfully tried in treatment of a variety of apparently unrelated dermatologic disorders. Meanwhile, thalidomide has been shown to possess a range of biologic actions, including inhibition of tumor necrosis factor alpha, possibly relevant to its clinical efficacy. Dermatologic disorders in addition to ENL in which thalidomide's effectiveness is well documented include aphthous stomatitis, discoid lupus erythematosus, actinic prurigo, Behcet's disease, and prurigo nodularis. More recently, the drug has been employed in dermatologic conditions associated with HIV infection. When used with safeguards to prevent teratogenicity and the drug's other major adverse effect, peripheral neuropathy, thalidomide may offer a good therapeutic option for many patients in whom other drug therapies have proven inadequate. PMID- 9859908 TI - Parvovirus B19-associated interstitial lung disease, hepatitis, and myositis. PMID- 9859911 TI - New antiviral agents for dermatologic disease. AB - The topic of antiviral agents for dermatologic disease is an area that is rapidly changing and expanding as our knowledge of herpes viruses, HIV, and human papilloma viruses continues to expand. New drugs are continuing to make therapy for these viruses more efficacious day by day. This review will discuss the recent advances in antiviral therapy, new therapies currently being developed or studied, and the future of antiviral vaccines. PMID- 9859912 TI - Ascomycins in dermatology. AB - Ascomycin derivatives represent a novel class of anti-inflammatory macrolactams that are under development for the treatment of skin diseases. The main biological effect of ascomycins is a blockage of the synthesis of both Th1- and Th2-type cytokines in target cells. SDZ ASM 981 is the most advanced ascomycin derivative under development. It has high antiinflammatory activity in animal models of allergic contact dermatitis and does not induce skin atrophy. Topical application of SDZ ASM 981 has been shown to be effective in atopic dermatitis (AD) and allergic contact dermatitis. Clinical studies using semi-occlusive conditions have also shown effectiveness in psoriasis. SDZ ASM 981 holds promise in overcoming the drawbacks of topical corticosteroids and studies are ongoing to further investigate its efficacy and safety in the treatment of inflammatory skin diseases. PMID- 9859913 TI - Pharmacological treatment of wounds. AB - Systemic treatment of patients with wounds can be directed at several physiological aspects of healing during the consecutive phases of tissue repair. Many approaches to pharmacological treatment have been tested in vitro, in animal experiments, or in clinical studies. Investigators and clinicians focus on treating underlying metabolic, infectious, inflammatory, or hemorrheological diseases and their complications. Drug treatment is often adjuvant or complementary to other measures such as compression, surgical intervention, reconstruction, or reopening procedures. This compounding fact can render interpretation of the results more difficult. As systemic treatment is not always without side-effects; local wound management is an option to consider. Topically applied growth factors certainly have the potential to influence the healing process. An indirect way of providing growth factors to wounds and chronic ulcers can be achieved by the application of grafts, cultured keratinocytes, and skin substitutes. Modulation of angiogenesis during wound healing is a recent target for research and treatment. Future reviews probably also will include genetic engineering methods for influencing wound healing. PMID- 9859914 TI - Recent advances in sun protection. PMID- 9859915 TI - Novel agents for the treatment of alopecia. AB - Recent approval in the United States of two new products, Propecia (Merck Co, Rahway, NJ) and Rogaine Extra Strength 5% (Pharmacia & UpJohn Co, Kalamazoo, MI), indicated in men to promote scalp hair growth, have added a new dimension to treatment options offered by physicians in treating androgenetic alopecia (AGA). The search for new and effective agents to treat many different hair loss problems has been intensified by the increase in hair biology research taking place worldwide, from university academic institutions to the pharmaceutical companies. All have a desire to profit from marketing such drugs that have been termed, "cosmeceuticals". Millions of men and women of every race suffer from various forms of alopecia, the most common being AGA where the target tissue active androgen, 5 alpha-dihydrotestosterone (DHT) aggravates genetically programmed scalp hair follicles that results in short, fine, miniaturized hairs. Currently available to treat alopecia are drugs indicated for other disease processes because no other agents are accessible; some have severe side-effects and many are minimally effective. These prescription drugs were not originally indicated for alopecia and have not been adequately tested in controlled clinical trials to assess for efficacy, safety, and toxicity. These agents continue to be used clinically to treat patients with various forms of alopecia. As a result, a variety of new agents are emerging in the patient application process to gain protection and approval specifically for various forms of alopecia. This report reviews the most recently approved products, some of the more promising compounds in clinical trial development, as well as those in the over-the-counter (OTC) "natural" treatments category. PMID- 9859916 TI - Alternative medicine and dermatology. AB - Because of increasing interest in the treatment and prevention of disease using nonconventional modalities, particularly in Western countries, it is important for practitioners of traditional Western medicine to remain open-minded about the use of alternative treatments. If the patient perceives the physician to be disapproving of the use of alternative treatments, she may not divulge the use of such treatments to the physician, even though alternative treatments can lead to adverse effects and to drug-herb interactions. The demographics and the reasons why patients seek alternative treatment are discussed. The scientific literature on the use of herbal and physical modalities is reviewed. Because of the large body of literature on the use of herbal remedies, the emphasis is on the current, most popular herbs in use by the general population, as well as on herbs used specifically for dermatologic disease. PMID- 9859917 TI - Quality of life assessments in dermatology. AB - This review describes the many ways in which skin disease can adversely affect the quality of life (QOL) of patients with skin disease. Measurement of this impact is required for clinical and health service research, and may be valuable in clinical practice and in the evaluation of new drug therapy. Methods of measuring QOL in dermatology are described. These include general health measures, dermatology specific measures such as the Dermatology Life Quality Index (DLQI), the Dermatology Quality of Life Scales (DQOLS), the Dermatology Specific Quality of Life (DSQL), and Skindex, and disease specific measures such as the Psoriasis Disability Index (PDI), the Psoriasis Life Stress Inventory (PLSI), and the Acne Disability Index (ADI). Instruments used for measuring QOL in children (the Children's Dermatology Life Quality Index (CDLQI)) and for measuring the impact of atopic dermatitis on the families of affected children (the Dermatitis Family Impact (DFI) questionnaire) are described. Reasons are given for the use of such measures in clinical practice. PMID- 9859918 TI - Pathogenesis of atherosclerosis: a possible relation to infection. AB - Atherosclerosis is the main underlying cause of coronary heart disease, which in turn is the most common cause of death in the industrialized world. An acute event in coronary heart disease is typically precipitated by thrombosis occurring at the site of atherosclerotic plaque disruption. Atherosclerotic plaques consist of a fibrous cap overlying a lipid-rich core. Many cell types are involved in their formation, including platelets, endothelial cells, activated monocytes, macrophages derived from monocytes and smooth muscle cells. The currently accepted hypothesis is that atherosclerosis develops as a response to injury and that it is primarily a chronic inflammatory condition. The endothelium plays an important role in regulating vascular blood flow and it is now apparent that endothelial dysfunction is an important contributor to the pathogenesis of atherosclerosis. There is growing evidence that infection may be a risk factor for atherosclerosis and myocardial infarction. Numerous studies have reported associations between human coronary heart disease (CHD) and bacterial and viral infections. At present, interest is focused on the potential aetiological role of C. pneumoniae which has been repeatedly identified, using various diagnostic techniques, in atherosclerotic lesions. There is also increasing seroepidemiological evidence of the association between C. pneumoniae and CHD. The role of this organism in atherosclerosis may be analogous to that of chronic C. trachomatis infection in trachoma. PMID- 9859919 TI - Seroepidemiological evidence for an association between Chlamydia pneumoniae and atherosclerosis. AB - The respiratory pathogen Chlamydia pneumoniae has been implicated in the pathogenesis of coronary heart disease. Evidence for this includes the direct demonstration of the organism in atherosclerotic plaques and the results of seroepidemiological studies. Of the several serological tests available, the microimmunofluorescence (MIF) test is the most widely used and has become a 'gold standard' because of its specificity and selectivity. In spite of differences in serological criteria of infection and interpretation, the findings of various research groups who have used the MIF test have suggested a link between atherosclerotic cardiovascular disease and Chlamydia pneumoniae infection. Further research is now required in accordance with standard criteria and using pre-defined techniques to confirm the role of the organism in coronary heart disease. PMID- 9859920 TI - Animal models for Chlamydia pneumoniae infection. AB - Animal models are used extensively in the ongoing investigation of a possible causal link between Chlamydia pneumoniae infection and conditions such as asthma and cardiovascular disease. Respiratory infections have been studied in monkeys, while mouse and rabbit models have been used to study both respiratory and cardiovascular infections. The degree of disease induced in mice depends on the strain used, the virulence of the C. pneumoniae strain used, and the dose administered. A characteristic mononuclear pneumonitis occurs, although the infection is systemic and the agent is found outside the lungs, in the circulation, spleen and liver. The infective dose used in the model tends to produce persistent infection, with inflammation continuing after the agent can no longer be cultured from the lungs. In reinfected animals the titre of infective chlamydia in lungs is much diminished, but the inflammation can be quite marked. The continuous persistence of the agent can be demonstrated by polymerase chain reaction (PCR), or, in chronically infected animals, after immunosuppression with cortisone. New Zealand White (NZW) rabbits provide an experimental model, not only for lung infections, but also for C. pneumoniae-induced atherosclerosis. Three laboratories have now reported that after inoculation, plaques develop in the arterial walls of experimental animals on a normal diet. In addition, one laboratory has reported from their studies on atherosclerosis in apoE-deficient and normal mice, that the persistence of the agent in aortic walls could be seen. Further studies are needed to evaluate the effect of the strain of chlamydia and dosage used, the importance of reinfection, the effect of diet and the effect of antibiotic treatment in these models. PMID- 9859921 TI - Chlamydia pneumoniae in vascular tissue. AB - This review summarizes the evidence for the presence of Chlamydia pneumoniae organisms in atherosclerotic lesions. A total of 17 publications are mentioned concerning this topic. In all but two, both from the same group, evidence is presented by means of various techniques for the existence of the organisms in atherosclerotic tissue. Thus, they have been found in the aorta and in coronary, carotid, pulmonary, femoral and iliac arteries. In the first study in the UK, the aorta, femoral and iliac arteries were found to be positive for C. pneumoniae. In a recent study on autopsy specimens undertaken in conjunction with South African investigators, 71% of atheromatous arteries were positive for C. pneumoniae compared with only 9% of non-atheromatous arteries. Interestingly, the organisms were detected in 67% of the vessels showing only early atherosclerotic lesions (fatty streaks). The relationship between the age of the subject and the probability of detecting C. pneumoniae organisms is discussed; the youngest subjects with arteries positive for these organisms were aged 15 and 20 years. It is concluded that the presence of chlamydiae within atherosclerotic lesions is now beyond doubt, but as yet there is no conclusive evidence for a causal role in atherosclerosis. PMID- 9859922 TI - Endovascular presence of Chlamydia pneumoniae DNA is a generalized phenomenon in atherosclerotic vascular disease. AB - The common respiratory pathogen Chlamydia pneumoniae has been implicated in the pathogenesis of coronary artery disease and acute myocardial infarction. In order to verify the endovascular presence of potentially viable chlamydia by detection of genomic DNA, we examined atherosclerotic arteries from various vascular regions using a C. pneumoniae specific nested polymerase chain reaction (PCR). The samples were obtained during surgical revascularization procedures or at autopsy. Chlamydial DNA was detected in 51/238 (21%) atherosclerotic samples. A total of 17 non-atherosclerotic control samples were PCR-negative. Chlamydial presence was detected in 36/140 (26%) vascular samples obtained at coronary revascularization procedures, in 9/61 (15%) samples from carotid artery stenosis, 3/17 (18%) samples from the aorta, and 3/20 (15%) iliac artery samples. Histomorphological discrimination of infected and non-infected arterial samples was not possible. Antichlamydial IgG and IgM response as examined by microimmunofluorescence assay did not aid identification of individual endovascular infection. C. pneumoniae is present in a significant proportion of atherosclerotic arteries. Its occurrence in atheromatous plaques is not limited to coronary arteries and may be considered indicative of an infectious component in atherosclerosis. However, it remains unclear whether chlamydia actually initiates atherosclerotic injury, facilitates its progression, or merely colonizes pre-existing atheromata. PMID- 9859923 TI - Chlamydia pneumoniae: inflammation and instability of the atherosclerotic plaque. AB - Coronary heart disease remains the most common cause of death in industrialized countries. Although atherosclerosis is generally asymptomatic in the early stages, progressive plaque development leads to arterial stenosis which is characterized by angina and may eventually lead to unstable angina, myocardial infarction and cardiac death. Evidence that the coagulation cascade is activated during acute coronary events has justified the use of antithrombotic agents such as aspirin, heparin and low molecular weight heparin (LMWH) in the standard management of acute coronary syndromes. The inflammatory process is also known to play a significant role in the pathogenesis of atherosclerosis, resulting in a cycle of continued inflammatory cell activation and ongoing cell recruitment. As the human leukocyte-associated antigen (HLA) system plays a key role in the regulation of the inflammatory process, the expression of HLA antigens in patients with symptomatic coronary heart disease has been investigated. These studies have demonstrated a relationship between the major histocompatibility complex (MHC) class II expression and the most severe pattern of angina refractory to conventional therapy, within the framework of a chronic infectious disease. A number of studies have documented an association between coronary heart disease and the presence of high titres of antibodies to Chlamydia pneumoniae, and this organism has been implicated in plaque instability. Such findings have stimulated interest in the role of C. pneumoniae in the pathogenesis of coronary heart disease, with a view to developing novel and effective treatment approaches. The ROXIS study showed a lower incidence of acute ischaemic events in patients with unstable angina treated with an antichlamydial antibiotic, roxithromycin. PMID- 9859924 TI - Genetics of Hb F/F cell variance in adults and heterocellular hereditary persistence of fetal hemoglobin. AB - Hb F and F cell values in normal adults vary considerably with a continuous distribution that is substantially skewed to the right implicating a polygenic influence. The high values of Hb F and F cells are transmitted in the condition referred to as heterocellular hereditary persistence fetal hemoglobin which should be regarded as a multifactorial quantitative trait, quite distinct from the classical pancellular hereditary persistence of fetal hemoglobins. Several factors have been shown to influence F cell/Hb F levels in normal adults including age, gender, genetic determinants linked and unlinked to the beta globin locus on chromosome 11p. Two trans-acting quantitative trait loci for F cell variance have been mapped, one on 6q and the other on Xp, with at least one other implicated. As an initial step towards hunting for the other quantitative trait loci we have carried out a preliminary analysis of F cell variance in 182 pairs of monozygotic and 373 pairs of dizygotic twins. The correlation coefficient of F cell variance in monozygotic twins was 0.89, while that in the dizygotic twins was 0.51. Overwhelming evidence for a strong genetic component in the control of Hb F/F cell levels is provided by a heritability of 0.87. However, the role and extent of contribution from the quantitative trait loci on 6q and Xp are still not known. PMID- 9859925 TI - F cells by immunofluorescent staining of erythrocyte smears. PMID- 9859926 TI - F reticulocytes assay: a method to evaluate fetal hemoglobin production. AB - Production of fetal hemoglobin (Hb F) involves molecular as well as cellular aspects as, among red blood cells, it is restricted to a specific population referred as the F cells. Thus understanding the mechanisms involved in persistence or re-emergence of Hb F production in various inherited or acquired conditions requires the measurement of both Hb F and F cells. In addition, in disorders with a hemolytic component, including sickle cell disease (SCD), because of a probable preferential survival of F cells as compared to non-F cells, the true parameter of F cell production is the F reticulocyte count. The F cells/F reticulocytes ratio then selectively reflects this preferential survival. Here we describe an original immunofluorescence microscopy assay that permits the simultaneous measurement of F cells and F reticulocytes. For this assay to be widely usable, we chose to use commercially available monoclonal antibodies. PMID- 9859927 TI - Cross evaluation of three flow cytometric F cell counting methods performed by different laboratories. AB - Three flow cytometric methods of counting F cells were evaluated in the settings of an external laboratory assessment scheme. The laboratories to participate with a different method were located in Oxford (method O), Athens (method A) and Jerusalem, (method J). Two monoclonal anti-gamma chain antibodies were used: monoclonal antibody produced by P. Beverley (Oxford) (BEV) and an antibody provided by Bioatlantic S.A.R.L. (France) (BIO). The specimens tested were mixtures in five predefined ratios of a sample with homozygous deltabeta thalassemia with 100% F cells with a sample with no F cells. A central independent laboratory prepared and distributed the aliquots (at room temperature) to the participating centers within 2 (O), 3 (A), and 6 (J) days. The performance of the three methods was evaluated by: 1) deviation indices, 2) relative accuracy, as percent difference of the counts from the target values, and 3) bias and linearity by linear regression of the counts versus the target values [parameters: slope (s), y-intercept (y), R squared (Rs), and F ratio]. The highest score of performance was obtained by method A with both monoclonal antibodies. PMID- 9859928 TI - Techniques for studying stimulation of fetal hemoglobin production in human erythroid cultures. AB - This report describes in detail the procedures for growing human erythroid cells in liquid culture for evaluating the potential of pharmacological agents to affect hemoglobin production. The procedure consists of two phases: an erythropoietin-independent phase in which peripheral blood mononuclear cells are first cultured in the presence of a combination of hemopoietic growth factors, but in the absence of erythropoietin, where early erythroid committed progenitors proliferate and differentiate into more mature progenitors. In the second phase, the latter cells, cultured in an erythropoietin-supplemented medium, continue to proliferate and mature into orthochromatic normoblasts and enucleated erythrocytes. This procedure produces large cultures of relatively pure and synchronized erythroid cell populations derived from normal donors or patients with beta hemoglobinopathies. The cultured cells recapitulate many aspects of erythropoiesis in vivo, including the donor's pattern of hemoglobin production (types and proportions). Tested compounds, at different concentrations, are added at different stages of the culture. The various types of hemoglobins and globin chains produced can be measured by high performance liquid chromatographic techniques and their cellular distribution analyzed by flow cytometry using fluorescently labeled antibodies against specific hemoglobins. This approach provides a screening system for compounds with potential therapeutic efficacy in patients with beta hemoglobinopathies. PMID- 9859929 TI - Determination of Hb F levels: the routine methods. AB - The two classical methods used to determine the level of Hb F in a hemolysate are herein described. Measurement of this hemoglobin fraction by resistance to alkali denaturation was the first technique introduced; it is today largely replaced by ion exchange high performance liquid chromatography. The limits, pitfalls, and advantages of the two methods are discussed. PMID- 9859930 TI - Gamma chain heterogeneity: determination of Hb F composition by perfusion chromatography. AB - The Ggamma:Agamma ratio is around 70:30 at the time of birth and usually 40:60 in the trace amounts of Hb F found in the adult. Changes in this ratio are observed in several hemoglobin disorders providing insights on the genetics and molecular pathophysiology of these diseases. Several techniques have been proposed to measure the Ggamma:Agamma ratio. We here describe perfusion chromatography which is now in routine use in one of our laboratories. The method involves a high velocity flow of the mobile phase through a porous reversed phase chromatographic stationary bed and allows us to determine this parameter one order of magnitude faster than with conventional high performance liquid chromatography. PMID- 9859931 TI - Indices of membrane alterations in beta-thalassemic erythrocytes. AB - Analytical protocols for the study of thalassemic erythrocyte membrane alterations are described. Denatured hemoglobin derivatives and aggregated band 3 are separated from detergent membrane extracts by gel-filtration as high molecular-weight aggregates and quantitated spectrophotometrically. Membrane bound, low-molecular-weight iron is measured on SDS-solubilized ghosts by a ferrozine-based colorimetric test. We adapted these methods for microscale preparation and analysis of erythrocyte ghosts in order to have suitable tools to estimate oxidative membrane damage in human samples. Data from 11 beta thalassemia intermedia patients and from 10 normal controls are reported as an example of the application of these methods. PMID- 9859932 TI - Red blood cell indices, cation content, and membrane cation transports. AB - In sickle cell disease, in the homozygous state, the increased heterogeneity of erythrocytes results mainly from membrane defects secondary to Hb S polymerization and the increased survival of F cells. The density distribution curve, using phthalate esters or the red blood cell indices measured with the H*3 system, are useful methods for the hematological follow-up of patients under specific therapies. The methods evaluating the red blood cell cation contents and the abnormal membrane potassium transport pathways are also described, in order to evaluate agents which can restore normal hemoglobin concentration and water content in dehydrated sickle cells. PMID- 9859933 TI - Globin chain synthesis analysis by high performance liquid chromatography in the screening of thalassemia syndromes. AB - We applied reversed phase high performance liquid chromatography for globin chain synthesis analysis in screening for beta-thalassemia. The alpha/non-alpha-globin chain synthesis ratios have been determined in alpha-, beta-, and deltabeta thalassemia carriers using the classical carboxymethyl cellulose chromatography as the reference method. Reversed phase high performance liquid chromatography is fast, accurate, and reproducible, and may be a suitable alternative for the traditional carboxymethyl cellulose chromatography. PMID- 9859934 TI - Molecular diagnosis and characterization of Hb Zurich [beta63(E7)His-->Arg]] carriers in a Kentucky family. PMID- 9859936 TI - Sheep Hb I variant, or beta13(A10)Gly-->Ser, in breeds from Corsica: detection by gene sequencing. PMID- 9859937 TI - Molecular differences in beta-thalassemia between the Spanish Mediterranean area and inland populations. PMID- 9859935 TI - Hb Tende [beta124(H2)Pro-->Leu]: a new variant with a moderate increase in oxygen affinity. PMID- 9859938 TI - Hb Koln [beta98(FG5)Val-->Met]: the first case found in a Chinese family. PMID- 9859939 TI - Measurements of human breast cancer using magnetic resonance spectroscopy: a review of clinical measurements and a report of localized 31P measurements of response to treatment. AB - A review of the literature has shown that in human breast tumours, large signals from phosphomonoesters (PME) and phosphodiesters (PDE) are evident. In serial measurements in 19 patients with breast cancer, a decrease in PME was significantly associated with a stable or responding disease (p = 0.017), and an increase in PME was associated with disease progression. Extract studies have shown PME to comprise of phosphoethanolamine (PEth) and phosphocholine (PCho), with the PEth to PCho ratio ranging from 1.3 to 12. The PCho content of high grade tumours was found to be higher than low grade tumours. In some animal models, changes in PCho have been shown to correlate with indices of cellular proliferation, and spheroid studies have shown a decrease in PCho content in spheroids with smaller growth fractions. A serial study of 25 patients with advanced primary breast tumours undergoing hormone, chemotherapy or radiotherapy treatments, showed that in this heterogenous group there were significant changes in metabolites that were seen during the first 3 weeks (range 2-4 weeks) of treatment, that correlated with volume change over this period, employed here as a measure of response. Changes in PME (p = 0.003), total phosphate (TP) (p = 0.008) and total nucleoside tri-phosphate (TNTP) (p = 0.02) over 3 (+/-1) weeks were significantly associated with response, as were the levels of PME (p<0.001), PDE (p = 0.01), TP (p = 0.001) and TNTP (p = 0.007) at week 3 (+/-1). PME at week 3 (+/-1) was also significantly associated with the best volume response to treatment (p = 0.03). A reproducibility analysis of results from the observation of normal breast metabolism in four volunteers showed a mean coefficient of variation of 25%, after correcting for changes resulting from the menstrual cycle. Reproducibility studies in four patients with breast cancer showed a mean coefficient of variation of 33%, with the reproducibility being better in patients measured on different days (difference in TP was -6%) compared with those measured on the same day (difference in TP was -29%). PMID- 9859940 TI - Phosphorus MR spectroscopy in the treatment of human extremity sarcomas. AB - The application of 31P MR spectroscopy in the characterization and treatment of malignant human extremity tumors is reviewed and placed in the perspective of results obtained in murine sarcomas. Despite the now widespread acquisition of gradient localized spectral maps, the low spatial resolution that can be achieved at 1.5 or 2 T with 31P MRS, greatly limits its use in the study of tumor heterogeneity. The potential of 31P MRS is in the evaluation and monitoring of large inoperable extremity tumors. There are early spectral changes in human extremity sarcomas monitored after therapy, and recent studies have shown that the 31P MR spectra measured before treatment, and the changes in phosphate metabolites measured shortly thereafter, correlate with the clinical response after 2 or 3 months. Larger clinical studies are needed to confirm whether correlations of, for instance, pretreatment tumor pH with necrosis at resection and Pi decrease with tumor regression, can be used as a predictive test for clinical response. PMID- 9859941 TI - Metabolic changes underlying 31P MR spectral alterations in human hepatic tumours. AB - Magnetic resonance spectroscopy (MRS) remains the technique of choice for observing tumour metabolism non-invasively. Although initially 31P MR spectroscopy showed much promise as a non-invasive diagnostic tool, studies of a wide range of hepatic tumours have conclusively shown that this technique cannot be utilized to distinguish between different tumour types. This lack of specificity and sensitivity appears to be a consequence of the fact that hepatic tumours develop with a range of modalities and not as a single abnormal disease process, and also because of the limited availability of MR detectable metabolic markers. This has led, in recent years, to a re-evaluation of the role of 31P MR spectroscopy, re-emerging as a non-invasive tool to follow the efficacy of the treatment regime. Furthermore, since the principal changes observed in tumours by 31P MRS appear to be an elevation in the concentration of phosphorylcholine (PCho) and phosphoethanolamine (PEth), new research using a combination of MRS and tissue culture of cell lines which carry a combination of known inducible oncogenes, are helping to elucidate some of the metabolic pathways that give rise to these metabolic alterations. PMID- 9859942 TI - Feasibility study of lactate imaging of head and neck tumors. AB - A proton spectroscopic imaging sequence was used to investigate the feasibility of lactate imaging in head and neck tumors. The sequence employs a two-shot lactate editing method with inversion recovery for additional lipid suppression, and a restricted field of view to suppress motion artifacts. Variations in acquisition parameters and two different receive coils were investigated on twelve patients. Elevated lactate was detected in three patients, no lactate was observed in seven patients, and two studies were inconclusive because of severe motion or inhomogeneity artifacts. Best results were obtained with an anterior/posterior neck coil at a 288 ms echo time (TE). PMID- 9859943 TI - NMR studies of phospholipid metabolism in hepatic lymphoma. AB - The detection of lymphomatous infiltration of the liver has implications for the staging and treatment of this disease. Our studies of patients with Hodgkin's disease and non-Hodgkin's lymphoma suggest that the involvement of the liver could be detected by 31P nuclear magnetic resonance (NMR) spectroscopy as an increase in the phosphomonoester/ATP and phosphomonoester/Pi ratios in the liver spectra in vivo. Studies of extracts of lymphomatous lymph nodes and of the lymphomatous mouse liver, showed that phosphoethanolamine was largely responsible for the increase in the phosphomonoester (PME) signal. This compound is involved in phospholipid metabolism, as a precursor and breakdown product of phosphatidylethanolamine. The kinetics of the synthesis of phosphatidylethanolamine from [13C2]ethanolamine were studied using 13C NMR spectroscopy. The increase in phosphoethanolamine in the lymphomatous liver was not found to be due to increased flux through the synthetic pathway to phosphatidylethanolamine, nor was it due to increased availability of ethanolamine. PMID- 9859945 TI - Current awareness in NMR in biomedicine. PMID- 9859944 TI - Non-invasive 19F-NMRS of 5-fluorouracil in pharmacokinetics and pharmacodynamic studies. AB - Knowledge of the exact dose and rate at which an antitumor agent is delivered to its target site is postulated to be crucial to proper patient management. It is now possible to obtain such information using non-invasive 19F-NMRS (nuclear magnetic resonance spectroscopy) following the administration of 5-fluorouracil (5-FU). We have performed such studies in 103 patients with breast, colorectal and other tumors. Measurable 19F signals were detected in 99 of these patients (92.5%). Estimation of the tumoral t1/2 of 5-FU in these patients revealed that 51 of them (51.5%) exhibited a tumoral t1/2 greater than 20 min, a value we had characterized as indicating drug trapping in the tumor. Of these patients, 46 who received regimen bolus 5-FU 600 mg/m2 with leucovorin for their treatment have been evaluated. In these patients, the association between trapping and response remains very high (p<.000001). None of the non-trappers responded to chemotherapy, whereas 70% of the evaluable trappers responded. Details are presented here on the methodology of NMRS data acquisition and on their pharmacokinetic analysis. The potential mechanisms underlying the trapping effect appear to be predicated primarily on transport processes. Suggestions are presented on how such pharmacokinetic imaging studies may extend both our understanding of the mechanism of action of 5-FU, and how they could be used to optimize patient treatment. PMID- 9859946 TI - Knowledge and perception of fall-related risk factors and fall-reduction techniques among community-dwelling elderly individuals. AB - BACKGROUND AND PURPOSE: Falls are a major cause of death and disability among older adults. Fall-related knowledge among community-dwelling elderly individuals, however, is unknown. The purposes of this study were to assess the perception of falling as a health problem, to determine the perceived importance of fall-related risk factors, and to identify personal characteristics and attitudes associated with perceived importance of fall-related risk factors among community-dwelling elderly people. SUBJECTS AND METHODS: Resident council meeting attendees in 3 apartment buildings for elderly persons completed a self administered survey (N=120). Items assessed factors influencing falls by the respondents (n=52) or falls among elderly persons (n=68). The data were analyzed by chi-square test, analysis of variance, and linear regression. RESULTS: Falling during the previous month was reported by 10.1% of the subjects. Eighty-six percent of the subjects considered falling to be a preventable health problem and viewed falling as a moderately important concern compared with other health concerns. Exterior environmental factors such as pavement conditions and handrails were perceived as most likely to cause falls. The perceived personal likelihood of falling due to these risk factors was markedly lower compared with the perceived risk for other elderly persons. CONCLUSION AND DISCUSSION: The community-dwelling elderly individuals studied considered falls to be preventable and understood the importance of fall-related risk factors, but they did not consider themselves to be susceptible to falling. PMID- 9859947 TI - Variations in posteroanterior stiffness in the thoracolumbar spine: preliminary observations and proposed mechanisms. AB - BACKGROUND AND PURPOSE: Evaluation of posteroanterior (PA) movement in the spine is commonly used in the clinic, but little is known about the mechanisms involved. The purposes of this study were to examine variations in PA stiffness along the thoracolumbar spine and to investigate possible factors that might determine the pattern of stiffness. SUBJECTS: Twenty-one pain-free volunteers (10 male, 11 female), aged 18 to 41 years (mean age=26.6, SD=7.5), participated. METHODS: Posteroanterior stiffness was measured at 5 locations (L4, L1, T10, T7, T4), together with various subject characteristics. RESULTS: Mean PA stiffness varied among locations, with the greatest stiffness at L4 (13.3 N/mm) and the lowest stiffness at L1 (10.4 N/mm). A relatively small, but important, proportion (22% or less) of the variance in stiffness data at some vertebral levels was accounted for by the variables describing subject characteristics. CONCLUSION AND DISCUSSION: Posteroanterior stiffness varies along the spine in a manner consistent with the nature of support for the spine. The observed pattern of variation of PA stiffness along the spine appears to be influenced by some factors other than those relating to the spine. PMID- 9859948 TI - A comparison of productivity and learning outcome in individual and cooperative physical therapy clinical education models. AB - BACKGROUND AND PURPOSE: This program evaluation was designed to evaluate productivity and the learning processes used during individual and cooperative clinical education experiences. SUBJECTS AND METHODS: Clinical instructors (n=23) and senior students (n=20) at the bachelor's degree level who were engaged in an individual learning experience at an Australian school provided workload productivity data on their daily patient care, administration, and teaching activities. An evaluation of the teaching and learning processes was conducted via questionnaire at the end of the experience. This same information was provided by a group of clinical instructors (n=8) and senior students (n=16) who were engaged in a cooperative learning experience. RESULTS: Clinical instructors in both learning experiences had to reduce their normal levels of productivity to supervise the students. The amount of patient care provided by students, however, compensated for this reduction in clinical instructor productivity. The extent of productivity gains in areas other than patient care were greater for the cooperative learning experience. Clinical instructors and students rated the individual and cooperative learning experiences similarly, although the students rated 3 particular learning processes more highly in the cooperative learning experience. CONCLUSION AND DISCUSSION: Advantages with respect to patient care, teaching, and administrative productivity were identified for each learning experience. From the perspective of the students, the cooperative learning experience appeared to provide additional educational benefits. PMID- 9859949 TI - Physical therapist management of lymphedema following treatment for breast cancer: a critical review of its effectiveness. AB - The purpose of this review is to analyze the research literature that has examined the effectiveness of physical therapy in the management of lymphedema following treatment for breast cancer. Thirteen studies met the criteria for experimental research, which were then categorized according to Sackett's levels of evidence. One study was graded at level II, 5 studies were graded at level III, and the remaining 7 studies were graded at level V. One grade B recommendation and 6 grade C recommendations were developed from the levels of evidence. The 13 selected studies were also graded according to 6 criteria to evaluate scientific rigor. Clinical recommendations and future research directions are provided. PMID- 9859950 TI - Physical therapy for a patient through six stages of amyotrophic lateral sclerosis. AB - BACKGROUND AND PURPOSE: This case report describes the use of Sinaki and Mulder's approach to staging amyotrophic lateral sclerosis (ALS) and functional outcome measures in designing a treatment program for a 59-year-old woman with ALS. CASE DESCRIPTION AND OUTCOMES: As the patient progressed from stage I through stage VI, over 12 months, the physical therapy goals changed from optimizing remaining function, to maintaining functional mobility, and finally to maximizing quality of life. DISCUSSION: Disease staging and the use of functional outcome measures provide a framework for physical therapy evaluation and treatment of patients with ALS throughout the disease process. Physical therapists can assist patients with ALS through the provision of education, psychological support, rehabilitation programs, and recommendations for appropriate equipment and community resources. PMID- 9859951 TI - An overview of the etiology, diagnosis, and treatment of Alzheimer disease. AB - Alzheimer disease (AD) is the most common form of dementia affecting elderly people. It is the fourth leading cause of death among adults in the United States, following heart disease, cancer, and stroke. The prevalence of AD increases with increasing age. An estimated 10% of people aged 65 years have this progressive, degenerative disease, and this percentage increases to 47.2% for people aged 85 years and older. An early-onset form of AD can affect individuals who are middle-aged, with the youngest documented case being that of a 28-year old. In the Framingham cohort, women with AD outnumbered men by a ratio of 2.8:1 for those aged 75 years or older. Undoubtedly, as our population continues to age, the increasing prevalence of AD will have an even greater impact on society than it does today. Approximately 4 million Americans have AD, and it is projected that the number will rise to 14 million by the middle of the next century. The financial impact of AD is staggering, with the average lifetime cost for an individual with AD exceeding $170,000. Although the majority of individuals with AD are cared for by family and friends at home, individuals with AD constitute half of all nursing home residents. The average cost of a year of nursing home care for an individual with AD is $42,000, and this cost can exceed $70,000. The purpose of this article is to provide an overview of the etiology of AD, the tools used in the diagnosis of AD, and the treatment of individuals with AD. In addition, the clinical presentation of the various stages of AD is described, and the psychosocial implications of this disease are discussed. PMID- 9859952 TI - Certification in CDP. Complete decongestive physiotherapy. PMID- 9859953 TI - Program accreditation. PMID- 9859954 TI - Surveillance for waterborne-disease outbreaks--United States, 1995-1996. AB - PROBLEM/CONDITION: Since 1971, CDC and the U.S. Environmental Protection Agency have maintained a collaborative surveillance system for collecting and periodically reporting data that relate to occurrences and causes of waterborne disease outbreaks (WBDOs). REPORTING PERIOD COVERED: This summary includes data for January 1995 through December 1996 and previously unreported outbreaks in 1994. DESCRIPTION OF THE SYSTEM: The surveillance system includes data about outbreaks associated with drinking water and recreational water. State, territorial, and local public health departments are primarily responsible for detecting and investigating WBDOs and for voluntarily reporting them to CDC on a standard form. RESULTS: For the period 1995-1996, 13 states reported a total of 22 outbreaks associated with drinking water. These outbreaks caused an estimated total of 2,567 persons to become ill. No deaths were reported. The microbe or chemical that caused the outbreak was identified for 14 (63.6%) of the 22 outbreaks. Giardia lamblia and Shigella sonnei each caused two (9.1%) of the 22 outbreaks; Escherichia coli O157:H7, Plesiomonas shigelloides, and a small round structured virus were implicated for one outbreak (4.5%) each. One of the two outbreaks of giardiasis involved the largest number of cases, with an estimated 1,449 ill persons. Seven outbreaks (31.8% of 22) of chemical poisoning, which involved a total of 90 persons, were reported. Copper and nitrite were associated with two outbreaks (9.1% of 22) each and sodium hydroxide, chlorine, and concentrated liquid soap with one outbreak (4.5%) each. Eleven (50.0%) of the 22 outbreaks were linked to well water, eight in noncommunity and three in community systems. Only three of the 10 outbreaks associated with community water systems were caused by problems at water treatment plants; the other seven resulted from problems in the water distribution systems and plumbing of individual facilities (e.g., a restaurant). Six of the seven outbreaks were associated with chemical contamination of the drinking water; the seventh outbreak was attributed to a small round structured virus. Four of the seven outbreaks occurred because of backflow or backsiphonage through a cross-connection, and two occurred because of high levels of copper that leached into water after the installation of new plumbing. For three of the four outbreaks caused by contamination from a cross connection, an improperly installed vacuum breaker or a faulty backflow prevention device was identified; no protection against backsiphonage was found for the fourth outbreak. Thirty-seven outbreaks from 17 states were attributed to recreational water exposure and affected an estimated 9,129 persons, including 8,449 persons in two large outbreaks of cryptosporidiosis. Twenty-two (59.5%) of these 37 were outbreaks of gastroenteritis; nine (24.3%) were outbreaks of dermatitis; and six (16.2%) were single cases of primary amebic meningoencephalitis caused by Naegleria fowleri, all of which were fatal. The etiologic agent was identified for 33 (89.2%) of the 37 outbreaks. Six (27.3%) of the 22 outbreaks of gastroenteritis were caused by Cryptosporidium parvum and six (27.3%) by E. coli O157:H7. All of the latter were associated with unchlorinated water (i.e., in lakes) or inadequately chlorinated water (i.e., in a pool). Thirteen (59.1%) of these 22 outbreaks were associated with lake water, eight (36.4%) with swimming or wading pools, and one(4.5%) with a hot spring. Of the nine outbreaks of dermatitis, seven (77.8%) were outbreaks of Pseudomonas dermatitis associated with hot tubs, and two (22.2%) were lake-associated outbreaks of swimmer's itch caused by Schistosoma species. INTERPRETATION: WBDOs caused by E. coli O157:H7 were reported more frequently than in previous years and were associated primarily with recreational lake water. This finding suggests the need for better monitoring of water quality and identification of sources of PMID- 9859955 TI - Cardiovascular disease risk factors and preventive practices among adults--United States, 1994: a behavioral risk factor atlas. Behavioral Risk Factor Surveillance System State Coordinators. AB - PROBLEM/CONDITIONS: Cardiovascular disease (CVD), including coronary heart disease (CHD) and stroke, is the leading cause of death in the United States, and state rates of CVD vary by state and by region of the country. Several behavioral risk factors (i.e., overweight, physical inactivity, smoking, hypertension, and diabetes mellitus) and preventive practices (i.e., weight loss and smoking cessation) are associated with the development of CVD and also vary geographically. This summary displays and analyzes geographic variation in the prevalences of selected CVD risk factors. REPORTING PERIOD: 1994 (1992 for prevalence of hypertension). DESCRIPTION OF SYSTEM: The Behavioral Risk Factor Surveillance System (BRFSS) is a state-based random-digit-dialing telephone survey of noninstitutionalized adults aged > or =18 years; 50 states and the District of Columbia participated in BRFSS in 1994, and 48 states and the District of Columbia participated in 1992. METHODS: Several different analyses were conducted: a) analysis of state risk factor and preventive practice prevalences by sex and race (i.e., black and white); b) mapping; c) cluster analysis; d) correlations of state prevalence rates by sex and race; and e) regression of state risk factor prevalences on state CHD and stroke mortality rates. RESULTS: Mapping the prevalence of selected CVD risk factors and preventive health practices indicates substantial geographic variation for black and white men and women, as confirmed by cluster analysis. Data for blacks are limited by small sample size, especially in western states. Geographic clustering is found for physical inactivity, smoking, and risk factor combinations. Risk factor prevalences are generally lower in the West and higher in the East. White men and white women are more similar in state risk factor rates than other race sex pairs; white women and black women ranked second in similarity. State prevalences of physical inactivity and hypertension are strongly associated with state mortality rates of CVD. INTERPRETATION: Geographic patterns of risk factor prevalence suggest the presence (or absence) of sociocultural environments that promote (or inhibit) the given risk factor or preventive behavior. Because the risk factors examined in this summary are associated with CVD, further exploration of the reasons underlying observed geographic patterns might be useful. The BRFSS will continue to provide geographic data about cardiovascular health behaviors with a possible emphasis on more data-based small- area analyses and mapping. This will permit states to more adequately monitor trends that affect the burden of CVD in their regions and the United States. Mapping also facilitates the exploration of patterns of morbidity, health-care use, and mortality, as well as the epidemiology of risk factors. Finally, by identifying those segments of the population with high levels of these risk factors and lower levels of the preventive health practices, public health personnel can better allocate resources and target intervention efforts for the prevention of CVD. PMID- 9859956 TI - Implications of the kinetics of zidovudine in the pregnant baboon following oral administration. AB - Zidovudine (ZDV) therapy in pregnancy reduces mother-to-child transmission of HIV. The action of ZDV in the fetus is thought to be an important contributor to efficacy. Previous research in primates has demonstrated that continuous infusion of ZDV to the mother leads to sustained plasma concentrations in the fetus; however, it has not been determined what concentrations of ZDV are achieved in the fetus following oral administration. The pharmacokinetics of drug distribution to the fetus following oral administration of a 100-mg dose of ZDV to the mother are reported from 6 chronically catheterized baboons. The first order elimination half-life of ZDV from both the mother and fetus was approximately 1.2 hours. The area under the concentration-time curve for the fetus was 77% (r2 = 0.98; p < .001) that of the mother and the estimated peak drug levels in the fetus were 52% (r2 = 0.83; p < .01) those in the mother. The rapid transfer and short half-life of ZDV leads to a drug concentration-time profile that would not sustain levels in the fetus with dosing every 4 hours. After comparing these findings with existing data from pregnant and nonpregnant humans, it seems likely that current dose recommendations for ZDV in pregnancy would not maintain levels of the active intracellular metabolite of ZDV in all fetuses. This may explain in part the 8% failure rate of ZDV prophylaxis. The correlation between fetal and maternal plasma concentrations of ZDV would allow titration of dose based on maternal drug levels to achieve fetal levels within the therapeutic range. PMID- 9859957 TI - Prechallenge high neutralizing antibodies and long-lasting immune reactivity to gp41 correlate with protection of rhesus monkeys against productive simian immunodeficiency virus infection or disease development. AB - To investigate the protective efficacy of various gp130 vaccine preparations, rhesus monkeys were immunized with gp130 oligomers (O-gp130) or two different gp130-monomer preparations (M1-gp130; M2-gp130) and challenged with 50 MID50 of simian immunodeficiency virus (SIV)mac32H. Following challenge the control animals and all animals of the M1- and M2-gp130 group and 1 animal of the O-gp130 group were productively infected, whereas 3 animals of the O-gp130 group resisted the productive virus replication. The protection was correlated with high neutralizing antibodies and a long-lasting immune response to the transmembrane protein gp41. Whereas none of the O-gp130 animals had developed disease symptoms, 3 M1-gp130 animals, 1 M2-gp130 animal, and 2 control animals died as a result of AIDS within 18 months after challenge. Therefore, immunization with virion derived gp130 oligomers of SIVmac32H can confer protection against the productive infection with SIVmac32H and suppress the development of the AIDS-like disease. PMID- 9859958 TI - Safety and immunogenicity of HIV recombinant envelope vaccines in HIV-infected infants and children. National Institutes of Health-sponsored Pediatric AIDS Clinical Trials Group (ACTG-218). AB - Study objectives were to evaluate the safety and immunogenicity of three HIV recombinant glycoproteins in HIV-infected infants and children between 1 month and 18 years of age with asymptomatic (P-1) infection. Using Chiron rgp 120 (SF 2) 15 or 50 microg; MicroGeneSys rgp 160 (IIIB) 40 or 320 microg; Genentech rgp120 (MN) 75 or 300 microg; or adjuvant control (Alum or MF-59), children were randomized to a double-blind, placebo-controlled, dose-escalating study of vaccine administered intramuscularly at entry and 1, 2, 3, 4, and 6 months later. No adverse events were attributed to study vaccines. Between 30% and 56% of volunteers exhibited a lymphoproliferative response as defined in terms of stimulation index (SI) to vaccine antigens; 65% of vaccinees but none of placebo recipients exhibited moderate or strong responses after enzyme immunoassay to HIV specific antigens. CD4 cell counts and quantitative HIV culture did not differ significantly among vaccine and control groups, nor were differences found among groups in HIV disease progression. The rgp160 and gp120 subunit vaccines were safe and immunogenic in this population. PMID- 9859960 TI - Time course of antibody response to tetanus toxoid and pneumococcal capsular polysaccharides in patients infected with HIV. AB - The temporal course of the humoral immune response to T-cell-dependent and T-cell independent type 2 antigens was evaluated in HIV-infected patients. In all, 26 seropositive patients were vaccinated with tetanus toxoid and 23-valent pneumococcal vaccines; total IgG and IgG1 antibodies to tetanus toxoid (Ttox) and total IgG and IgG2 antibodies against 23 Streptococcus pneumoniae capsular antigens (PPS) were measured at baseline, 2 months, and 12 months after vaccination. For the Ttox, baseline levels of IgG1 (Ttox-IgG1) increased from 11.0 to 19.5 mg/L at 2 months postimmunization. Overall only 6 patients (23%) showed a significant response. At 12 months postvaccination, Ttox-IgG and T-tox IgG1 were significantly lower than baseline levels (Ttox IgG basal; 11.0 mg/L, 12 months; 0.8 mg/L, Ttox IgG1 baseline; 13.1 mg/L, Ttox IgG1 12 months; 2.4 mg/L) and in 10 patients, antibodies that fell below protective levels (0.6 mg/L). In contrast with PPS, a significant response was observed at 2 and 12 months (PPS IgG basal; 35.9 U/ml, 2 months; 151.4 U/ml, 12 months; 59.7 U/ml; PPS-IgG2 baseline 20.3 U/ml, 2 months; 113.2 U/ml, 12 months; 51.9 U/ml). Overall, 19 patients (76%) showed an immune response to pneumococcal polysaccharides antigens. Immunization with the Ttox T-cell-dependent antigen fails to elicit a significant immune response and may induce inhibition of antibody production in HIV-infected patients. In contrast, immunization with a T-cell-independent type 2 antigen can cause the pneumococcal polysaccharides to induce significant immune response in a high proportion of HIV-infected patients. PMID- 9859959 TI - Maternal and perinatal factors related to maternal-infant transmission of HIV-1 in the P2C2 HIV study: the role of EBV shedding. Pediatric Pulmonary and Cardiovascular Complications of Vertically Transmitted HIV-1 Infection (P2C2 HIV) Study Group. AB - The association of maternal and perinatal factors with mother-infant transmission of HIV-1 was examined in a prospective multicenter cohort of singleton live births to 508 HIV-1-infected women with children of known HIV-1 infection status (91 [18%] HIV-1-infected, 417 [82%] uninfected). From multivariate logistic regression, independent predictors of HIV-1 transmission included maternal CD4 percentage (CD4%) (odds ratio [OR] per 10% increase in CD4% = 0.70; p = .003), ruptured membranes <24 hours (OR = 3.15; p = .02), and maternal bleeding (OR = 2.90; p = .03), whereas maternal zidovudine (ZDV) use was marginally associated (OR = 0.60; p = .08). The associations of maternal urinary cytomegalovirus (CMV) shedding, oropharyngeal Epstein-Barr virus (EBV) shedding, and serology profiles during pregnancy with HIV-1 transmission were examined in the subset of mothers in whom the CMV and EBV measurements were available. Maternal EBV seropositivity, CMV shedding, and CMV seropositivity were 100% (279 of 279), 7% (16 of 229), and 92% (270 of 274), respectively. These rates did not differ between transmitting and nontransmitting mothers. In univariate analyses, maternal EBV shedding was higher among transmitting than nontransmitting mothers (40 of 49 [82%] compared with 154 of 226 [68%]; p = .06) and was independently associated with transmission in multivariate logistic analyses adjusting for CD4%, ruptured membranes, and ZDV use, with an OR of 2.45 (95% confidence interval (CI), 1.03 5.84; p = .04). This permits the conclusion that EBV shedding is associated with maternal-infant HIV-1 transmission, independent of CD4%. PMID- 9859961 TI - Decay of HIV-1 DNA in patients receiving suppressive antiretroviral therapy. AB - We have examined the effect of potent antiretroviral regimens on the latent reservoirs of HIV-1. The HIV-1 DNA in the peripheral blood mononuclear cells (PBMC) of 10 patients with undetectable plasma HIV-1 RNA (<20 copies/ml) who had received combination antiretroviral therapy was assayed every 12 weeks. No evidence of residual viral replication was found in the PBMC after 24 weeks of treatment. Although HIV-1 DNA remained detectable in all patients, it decreased significantly from 3.5 log copies/10(6) cells (range, 1.8-4.7 log copies/10(6) cells) to 2.3 log copies/10(6) cells (range, 0.6-3.1 log copies/10(6) cells) after 60 weeks of suppressive therapy. Analysis based on 6 patients who reached 60 weeks showed a slow decline with an estimated half-life of 40 weeks (range, 26 163 weeks). Genotypic analysis by sequencing the HIV-1 pol gene revealed no changes in the reverse transcriptase or protease coding regions after 48 to 60 weeks of therapy. The findings suggest that, in addition to potent antiretroviral regimens, new strategies must be developed to ensure eradication of the latent reservoir of provirus, and hence of the virus itself. PMID- 9859962 TI - A prospective study of community-acquired bloodstream infections among febrile adults admitted to Mulago Hospital in Kampala, Uganda. AB - Septicemia is a frequent cause of death in HIV-infected adults in developing countries. Additional prospective studies are needed to determine the etiology of bloodstream infections (BSI) in febrile HIV-infected adults and guide initial evaluation and treatment in this setting. We assessed the prevalence and etiology of community-acquired BSI among 299 consecutive febrile adult medical admissions to Mulago Hospital, Kampala, Uganda, over a 4-month period in 1997. The median age of our patients was 30 years, 159 (53%) were male, and 227 (76%) HIV-1 seropositive. Overall, prevalence of bacteremia or fungemia (1 patient) was 24%. Bacteremia was more frequent in HIV-infected than in uninfected patients (27% versus 15%, respectively; p = .04). Mycobacterium tuberculosis (n = 28), Streptococcus pneumoniae (n = 15) and Salmonella species (n = 13) were the most frequent isolates. All Salmonella and mycobacterial isolates were recovered from HIV-infected patients. Pneumococcal bacteremia was not associated with HIV seropositivity. M. avium complex and M. simiae were isolated from two HIV infected patients. The rate of mycobacteremia among febrile HIV-infected adults presenting for hospitalization was 13%. Bacteremia and disseminated tuberculosis are frequent causes of morbidity in febrile HIV-infected Ugandan adults. Initial empiric antibiotic coverage in this setting should be targeted toward the pneumococcus and gram-negative enteric bacilli, especially nontyphi Salmonella species. All patients presenting with chronic cough should be evaluated for tuberculosis. PMID- 9859963 TI - Nosocomial bloodstream infections in HIV-infected patients: attributable mortality and extension of hospital stay. AB - A 3-year prospective matched case-control study was performed to investigate the potential risk factors, prognostic indicators, extension of hospital stay, and attributable mortality of nosocomial bloodstream infections in HIV-infected patients. Matching variables were: age, gender, number of circulating CD4+ T lymphocytes, cause of hospital admission, hospitalization in the same ward within the 6 weeks of diagnosis of the case, and length of stay before the day of infection in the case. Eighty-four cases and 168 matched controls were studied. Nosocomial bloodstream infections complicated about 3 of 1000 hospital days per patient in the study period. With step-wise logistic regression analysis, the most important predictors for developing nosocomial bloodstream infections were: increasing value of Acute Physiology and Chronic Health Evaluation (APACHE II) score (p = .001) and use of central venous catheter (CVC) (p = .002). The excess of hospital stay attributable to nosocomial bloodstream infections was 17 days. The crude mortality rate was 43%. The attributable mortality rate was estimated to be 27% (95% confidence interval [CI] = 13%-48%). The estimated risk ratio for death was 3.91 (95% CI = 2.06-7.44). Multivariate analysis identified two prognostic indicators that were significantly associated with unfavorable outcome of bloodstream infections: number of circulating CD4+ T cells <100/mm3 (p = .002) and APACHE II score >15 (p = .01). Nosocomial bloodstream infections are more common in patients with advanced HIV disease. Important cofactors are high APACHE II score and use of CVC. These infections can cause an excess mortality and significantly prolong the hospital stay of HIV-infected patients. PMID- 9859964 TI - A randomized trial of N-acetylcysteine for prevention of trimethoprim sulfamethoxazole hypersensitivity reactions in Pneumocystis carinii pneumonia prophylaxis (CTN 057). Canadian HIV Trials Network 057 Study Group. AB - Hydroxylamine derivatives of sulfamethoxazole may be the reactive metabolites that cause adverse reactions to trimethoprim-sulfamethoxazole (TMP-SMX). The increased frequency of reactions observed in HIV-positive individuals is hypothesized to be due to systemic glutathione deficiency and a decreased ability to scavenge these metabolites. Two hundred and thirty-eight patients were randomized to receive or not receive N-acetylcysteine (3 g of the 20% liquid solution) 1 hour before each dose of TMP-SMX (trimethoprim 80 mg, sulfamethoxazole 400 mg) twice daily, which was initiated as primary Pneumocystis carinii pneumonia prophylaxis. Forty-five patients had to discontinue TMP-SMX within 2 months because of fever, rash, or pruritus including 25 of 102 patients (25%) who were receiving TMP-SMX alone and 20 of 96 patients (21%) who were randomized to TMP-SMX and N-acetylcysteine. The difference between treatment groups is 4% (95% confidence interval [CI]: -16%, +9%). No independent association was found with the hypersensitivity reaction and age, gender, race, HIV risk factor, prior AIDS, concurrent use of fluconazole, or baseline CD4. N acetylcysteine at a dose of 3 g twice daily could not be shown to prevent TMP-SMX hypersensitivity reactions in patients with HIV infection. PMID- 9859965 TI - HIV seroincidence and risk factors among patients repeatedly tested for HIV attending sexually transmitted disease clinics in the United States, 1991 to 1996. STD Clinic HIV Seroincidence Study Group. AB - To assess the incidence of HIV infection and risk factors associated with HIV seroconversion among patients attending clinics for sexually transmitted diseases (STD), medical record reviews were conducted in 12 clinics in 7 U.S. cities. The records of all patients who initially tested negative for HIV from 1991 through 1996 and who received at least one additional HIV test during the study period were reviewed. In each of 7 cities, 5 to 112 patients seroconverted. Of the 286 seroconverters identified in total, 53% (152 of 286) were heterosexual men and 28% (81 of 286) were women. HIV incidence rates among men who have sex with men (MSM) ranged by city from 0.81 to 7.0 new infections/100 person-years. Rates among heterosexual men and women ranged from 0.018 to 1.2 infections/100 person years. Multivariate analyses showed that drug use was associated with HIV seroconversion only among heterosexuals. Most new HIV infections in these clinics are being transmitted heterosexually and are associated with drug use. Nevertheless, MSM, particularly young MSM, are at greatest risk for HIV in this population: 1 of 47 seroconvert/year. The effective use of targeted prevention efforts depends upon the continued ability to monitor the incidence of HIV infection. PMID- 9859966 TI - Seroprevalence of HTLV-I and HTLV-II among a cohort of HIV-infected women and women at risk for HIV infection. Women's Interagency HIV Study. AB - OBJECTIVES: To determine the seroprevalence of, and risk factors for, HTLV-I and HTLV-II infection among HIV-infected women and women at high risk for HIV infection. DESIGN: Cross-sectional analysis of baseline data for women enrolled in the prospective Women's Interagency HIV Study (WIHS). METHODS: From October 1994 through November 1995, 2657 women from five metropolitan areas in the United States (Chicago, Los Angeles, New York City [two sites], Northern California, and Washington DC) were enrolled in WIHS. An interview-based survey collected data on demographics, behavior, and medical history. HTLV-I and HTLV-II determinations were made using a combined HTLV-I/HTLV-II indirect immunofluorescent antibody (IFA) screening test, an IFA titration specificity test, and individual HTLV-I and HTLV-II confirmatory Western blots. Fisher's exact tests and logistic regression were used to determine univariate and multivariate independent predictors for HTLV-II infection. RESULTS: Of 2625 women enrolled in WIHS with confirmed HIV results, 2487 (95%) were tested for HTLV-I and HTLV-II. Of these, 241 (10%) were HTLV-II-seropositive and 13 (0.5%) were HTLV-I-seropositive. On multivariate analysis, independent predictors of HTLV-II infection included injection drug use (OR = 5.2; p < .001), black race (OR = 3.6; p < 0.001), age >35 years (OR = 3.3; p < .001) and a history of sex with a male injecting drug user (OR = 1.9; p < .001). Among women infected with HIV, the seroprevalence of HTLV-II was 11% compared with 6% for women at risk for HIV but not infected (p < .001). However, HIV was not an independent predictor of HTLV-II infection in multivariate analysis. CONCLUSIONS: This cross-sectional analysis confirms that HTLV-II is found commonly in HIV-infected women and uninfected women at risk for HIV in major urban areas throughout the United States and that HTLV-II is far more common than HTLV-I in these populations. Although injecting drug use is most strongly associated with HTLV-II infection, sexual transmission likely contributes to the high HTLV-II seroprevalence in this cohort. PMID- 9859967 TI - Preventing discrimination against volunteers in prophylactic HIV vaccine trials: lessons from a phase II trial. AB - CONTEXT: Preventive HIV vaccines can temporarily cause uninfected individuals to have positive results on HIV testing. As preparations are underway to mount larger efficacy trials, the social risks of trial participation should be studied. OBJECTIVE: To describe frequency of HIV testing and discrimination among participants in a preventive phase II HIV vaccine trial. PARTICIPANTS: 266 vaccine trial volunteers were eligible; 247 participated in a confidential survey. RESULTS: 63 volunteers (26% of respondents) reported 185 HIV tests during the prior 12 to 24 months; most tests were for other research studies, health care, insurance, incarceration, or employment. Only 5 volunteers reported having positive HIV test results. Volunteers reported 99 adverse social incidents or problems, 53 of which were related to the trial. The most common type of event occurred when volunteers disclosed their trial participation and were mistakenly presumed to be infected with HIV. Few reported difficulty obtaining insurance, job loss, and inadvertent disclosure of their participation in the trial. CONCLUSION: In this vaccine trial, few serious social harms were reported. Those who conduct HIV tests for insurance, employment, health care, or other reasons should be made aware that HIV vaccines can cause false-positive HIV test results. Those planning future trials must continue to provide needed support to volunteers. Social harms should be monitored with the same vigilance accorded to physical harms. PMID- 9859968 TI - Epidemiologic evidence for time variation in HIV infectivity. AB - Data from three epidemiologic studies of heterosexual transmission of HIV among monogamous couples are used to assess evidence for time variation in HIV infectivity, possibly related to varying levels of infectiousness following infection in the primary infected partner. Analyses are based on statistical techniques that account for the inherent incompleteness of exposure information from such studies, and that allow direct assessment of the hypotheses that infectivity varies with time since infection and across partnerships. Data include findings from 302 couples from the California Partners' Study and 51 and 31 couples, respectively, from two U.S. Center for Disease Control and Prevention (CDC)-sponsored studies of infection in partners of transfusion recipients. Results indicate weak evidence for higher infectivity following infection of the primary partner, decreasing to relatively lower levels from 2 to 10 years after. Although these findings are consistent with biologic observations of time variation in viral levels, other explanations of the observed pattern (e.g., heterogeneity of infectivity) are equally plausible, pointing out some inherent limitations of data from such studies. PMID- 9859969 TI - HTLV-I-associated myelopathy/tropical spastic paraparesis in Brazil: a nationwide survey. HAM/TSP Brazilian Study Group. AB - To study the epidemiology of HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) in Brazil, we conducted a nationwide survey between March 1994 and April 1995. Five centers from three regions of the country participated, enrolling 163 patients. Most patients came from the northeastern and southeastern regions (93.2%). Most enrollees were white women, 42.9% and 64.4%, respectively. The most common risk factors for infection included a history of venereal diseases (30.6%) and blood transfusion (21.6%). The median age at the beginning of the disease was 42 years. The main neurologic findings were spastic paraparesis, widespread brisk tendon jerks, bilateral Babinski's sign, and bladder dysfunction. Some interregional differences reached statistical significance. The ratio of females over males increased from south to north. In addition, in both southern and southeastern regions, whites prevailed, whereas in the northeast, mulattos predominated. This follows the normal distribution of the population in these regions. A significantly higher rate of venereal diseases was found in the southeast compared with the other regions studied. A history of intravenous drug use was more frequent among patients as the sample moves south. Finally, a fluctuating course of the disease was proportionally more frequent in the southern region. PMID- 9859970 TI - Simian T-lymphotropic virus type I infection among wild-caught Indonesian pig tailed macaques (Macaca nemestrina). AB - Evidence for the presence of simian T-lymphotropic viruses (STLV-I) was identified in live-caught pig-tailed macaques from two locations in southern Sumatra, Indonesia. Of 60 animals tested, 13.3% of the animals showed seroreactivity to HTLV-I/II enzyme-linked immunosorbent assay (ELISA) antigens. Of these, 75% showed indeterminate reactivity and 25% showed positive reactivity to HTLV-I/II Western blot antigens. Polymerase chain reaction (PCR) analysis of 6 of 8 seroreactive monkeys' peripheral blood mononuclear cell (PBMC) DNA showed production of proper size molecular weight product that hybridized specifically to an STLV-I tax gene-specific probe. Phylogenic analyses of tax gene fragment sequences from the PCR products of two samples, 930287 and 930306, indicated that these animals were infected with retroviruses related to those of the Asian STLV I clade. PMID- 9859971 TI - Effect of coinfection with hepatitis G virus on HIV disease progression in hemophilic men. PMID- 9859972 TI - HTLV-I/II antibodies among three different Indian groups from Paraguay. PMID- 9859973 TI - Patent on gene fragment sends researchers a mixed message...as Germany hesitates over Brussels directive. PMID- 9859974 TI - Allow cloning in embryo research, says UK report. PMID- 9859976 TI - Palaeontologists divided over 'stay at home' policy for fossils. PMID- 9859975 TI - US scientists may boycott AIDS congress. PMID- 9859977 TI - Animal rights activists turn the screw. PMID- 9859978 TI - Open up review system. PMID- 9859979 TI - Whose law for sharing research tools? PMID- 9859980 TI - Membrane fusion. Ready...aim...fire! PMID- 9859982 TI - Palaeontology. The face of Cinderella. PMID- 9859981 TI - A hydrogen-producing mitochondrion. PMID- 9859984 TI - The A to Z of DNA. PMID- 9859983 TI - Evolutionary biology. A plastic genome. PMID- 9859985 TI - Kendrew constructs; Geis gazes. PMID- 9859986 TI - A hydrogenosome with a genome. PMID- 9859987 TI - p53 polymorphism and risk of cervical cancer. PMID- 9859988 TI - p53 polymorphism and risk of cervical cancer. PMID- 9859989 TI - p53 polymorphism and risk of cervical cancer. PMID- 9859990 TI - Defining the functions of trans-SNARE pairs. AB - The homotypic fusion of yeast vacuoles includes a 'docking' step, which we show here to consist of two sequential reactions: a reversible 'tethering' mediated by the GTPase Ypt7, and 'SNARE pairing', in which SNARE proteins from opposite membranes form a complex in trans. The function of this trans-SNARE complex must be transient, as the complex can be disassembled by excess Sec18 in the presence of Sec17 and ATP without influencing the fusion rate. These data indicate that SNARE pairing may transiently signal to downstream factors, leading to fusion. PMID- 9859991 TI - Selective sweep of a newly evolved sperm-specific gene in Drosophila. AB - The pattern of genetic variation across the genome of Drosophila melanogaster is consistent with the occurrence of frequent 'selective sweeps', in which new favourable mutations become incorporated into the species so quickly that linked alleles can 'hitchhike' and also become fixed. Because of the hitchhiking of linked genes, it is generally difficult to identify the target of any putative selective sweep. Here, however, we identify a new gene in D. melanogaster that codes for a sperm-specific axonemal dynein subunit. The gene has a new testes specific promoter derived from a protein-coding region in a gene encoding the cell-adhesion protein annexin X (AnnX), and it contains a new protein-coding exon derived from an intron in a gene encoding a cytoplasmic dynein intermediate chain (Cdic). The new transcription unit, designated Sdic (for sperm-specific dynein intermediate chain), has been duplicated about tenfold in a tandem array. Consistent with the selective sweep of this gene, the level of genetic polymorphism near Sdic is unusually low. The discovery of this gene supports other results that point to the rapid molecular evolution of male reproductive functions. PMID- 9859992 TI - Ca2+/calmodulin signals the completion of docking and triggers a late step of vacuole fusion. AB - The basic reaction mechanisms for membrane fusion in the trafficking of intracellular membranes and in exocytosis are probably identical. But in contrast to regulated exocytosis, intracellular fusion reactions are referred to as 'constitutive' as no final Ca2+-dependent triggering step has been observed. Although transport from the endoplasmic reticulum to the Golgi apparatus in the cell depends on Ca2+, as does endosome fusion and assembly of the nuclear envelope, it is unclear whether Ca2+ triggers these events. Membrane fusion involves several subreactions: priming, tethering and docking. Proteins that are needed for fusion include p115, SNAPs, NSF, SNAREs and small GTPases, which operate in these early reactions, but the machinery that catalyses the final mixing of biological membranes is still unknown. Here we show that Ca2+ is released from the vacuolar lumen following completion of the docking step. We have identified calmodulin as the putative Ca2+ sensor and as the first component required in the post-docking phase of vacuole fusion. Calmodulin binds tightly to vacuoles upon Ca2+ release. Unlike synaptotagmin or syncollin in exocytosis, calmodulin does not act as a fusion clamp but actively promotes bilayer mixing. Hence, activation of SNAREs is not sufficient to drive bilayer mixing between physiological membranes. We propose that Ca2+ control of the latest phase of membrane fusion may be a conserved feature, relevant not only for exocytosis, but also for intracellular, 'constitutive' fusion reactions. However, the origin of the Ca2+ signal, its receptor and its mode of processing differ. PMID- 9859993 TI - Control of apoptosis and mitotic spindle checkpoint by survivin. AB - Progression of the cell cycle and control of apoptosis (programmed cell death) are thought to be intimately linked processes, acting to preserve homeostasis and developmental morphogenesis. Although proteins that regulate apoptosis have been implicated in restraining cell-cycle entry and controlling ploidy (chromosome number), the effector molecules at the interface between cell proliferation and cell survival have remained elusive. Here we show that a new inhibitor of apoptosis (IAP) protein, survivin, is expressed in the G2/M phase of the cell cycle in a cycle-regulated manner. At the beginning of mitosis, survivin associates with microtubules of the mitotic spindle in a specific and saturable reaction that is regulated by microtubule dynamics. Disruption of survivin microtubule interactions results in loss of survivin's anti-apoptosis function and increased caspase-3 activity, a mechanism involved in cell death, during mitosis. These results indicate that survivin may counteract a default induction of apoptosis in G2/M phase. The overexpression of survivin in cancer may overcome this apoptotic checkpoint and favour aberrant progression of transformed cells through mitosis. PMID- 9859994 TI - Calcium promotes cell survival through CaM-K kinase activation of the protein kinase-B pathway. AB - The protection against apoptosis provided by growth factors in several cell lines is due to stimulation of the phosphatidylinositol-3-OH kinase (PI(3)K) pathway, which results in activation of protein kinase B (PKB; also known as c-Akt and Rac) and phosphorylation and sequestration to protein 14-3-3 of the proapoptotic Bcl-2-family member BAD. A modest increase in intracellular Ca2+ concentration also promotes survival of some cultured neurons through a pathway that requires calmodulin but is independent of PI(3)K and the MAP kinases. Here we report that Ca2+/calmodulin-dependent protein kinase kinase (CaM-KK) activates PKB directly, resulting in phosphorylation of BAD on serine residue 136 and the interaction of BAD with protein 14-3-3. Serum withdrawal induced a three- to fourfold increase in cell death of NG108 neuroblastoma cells, and this apoptosis was largely blocked by increasing the intracellular Ca2+ concentration with NMDA (N-methyl-D aspartate) or KCl or by transfection with constitutively active CaM-KK. The effect of NMDA on cell survival was blocked by transfection with dominant negative forms of CaM-KK or PKB. These results identify a Ca2+-triggered signalling cascade in which CaM-KK activates PKB, which in turn phosphorylates BAD and protects cells from apoptosis. PMID- 9859995 TI - Decoupling of nucleotide- and microtubule-binding sites in a kinesin mutant. AB - Molecular motors require ATP to move along microtubules or actin filaments. To understand how molecular motors function, it is crucial to know how binding of the motor to its filamentous track stimulates the hydrolysis of ATP by the motor, enabling it to move along the filament. A mechanism for the enhanced ATP hydrolysis has not been elucidated, but it is generally accepted that conformational changes in the motor proteins occur when they bind to microtubules or actin filaments, facilitating the release of ADP. Here we report that a mutation in the motor domain of the microtubule motor proteins Kar3 and Ncd uncouples nucleotide- and microtubule-binding by the proteins, preventing activation of the motor ATPase by microtubules. Unlike the wild-type motors, the mutants bind tightly to both ADP and microtubules, indicating that interactions between the nucleotide- and microtubule-binding sites are blocked. The region of the motor that includes the mutated amino acid could transmit or undergo a conformational change required to convert the motor ATPase into a microtubule stimulated state. PMID- 9859996 TI - Identification of the receptor component of the IkappaBalpha-ubiquitin ligase. AB - NF-kappaB, a ubiquitous, inducible transcription factor involved in immune, inflammatory, stress and developmental processes, is retained in a latent form in the cytoplasm of non-stimulated cells by inhibitory molecules, IkappaBs. Its activation is a paradigm for a signal-transduction cascade that integrates an inducible kinase and the ubiquitin-proteasome system to eliminate inhibitory regulators. Here we isolate the pIkappaBalpha-ubiquitin ligase (pIkappaBalpha-E3) that attaches ubiquitin, a small protein which marks other proteins for degradation by the proteasome system, to the phosphorylated NF-kappaB inhibitor pIkappaBalpha. Taking advantage of its high affinity to pIkappaBalpha, we isolate this ligase from HeLa cells by single-step immunoaffinity purification. Using nanoelectrospray mass spectrometry, we identify the specific component of the ligase that recognizes the pIkappaBalpha degradation motif as an F-box/WD-domain protein belonging to a recently distinguished family of beta-TrCP/Slimb proteins. This component, which we denote E3RSIkappaB (pIkappaBalpha-E3 receptor subunit), binds specifically to pIkappaBalpha and promotes its in vitro ubiquitination in the presence of two other ubiquitin-system enzymes, E1 and UBC5C, one of many known E2 enzymes. An F-box-deletion mutant of E3RS(IkappaB), which tightly binds pIkappaBalpha but does not support its ubiquitination, acts in vivo as a dominant negative molecule, inhibiting the degradation of pIkappaBalpha and consequently NF-kappaB activation. E3RS(IkappaB) represents a family of receptor proteins that are core components of a class of ubiquitin ligases. When these receptor components recognize their specific ligand, which is a conserved, phosphorylation based sequence motif, they target regulatory proteins containing this motif for proteasomal degradation. PMID- 9859997 TI - Regulation of activity of the transcription factor GATA-1 by acetylation. AB - Modification of histones, DNA-binding proteins found in chromatin, by addition of acetyl groups occurs to a greater degree when the histones are associated with transcriptionally active DNA. A breakthrough in understanding how this acetylation is mediated was the discovery that various transcriptional co activator proteins have intrinsic histone acetyltransferase activity (for example, Gcn5p, PCAF, TAF(II)250 and p300/CBP. These acetyltransferases also modify certain transcription factors (TFIIEbeta, TFIIF, EKLF and p53). GATA-1 is an important transcription factor in the haematopoietic lineage and is essential for terminal differentiation of erythrocytes and megakaryocytes. It is associated in vivo with the acetyltransferase p300/CBP. Here we report that GATA-1 is acetylated in vitro by p300. This significantly increases the amount of GATA-1 bound to DNA and alters the mobility of GATA-1-DNA complexes, suggestive of a conformational change in GATA-1. GATA-1 is also acetylated in vivo and acetylation directly stimulates GATA-1-dependent transcription. Mutagenesis of important acetylated residues shows that there is a relationship between the acetylation and in vivo function of GATA-1. We propose that acetylation of transcription factors can alter interactions between these factors and DNA and among different transcription factors, and is an integral part of transcription and differentiation processes. PMID- 9859998 TI - Outcomes of primary phakic trabeculectomies without versus with 0.5- to 1-minute versus 3- to 5-minute mitomycin C. AB - PURPOSE: To compare the intraocular pressure and hypotony outcomes of primary phakic trabeculectomies with no mitomycin C (MMC), shorter MMC, and longer MMC exposure. METHODS: We evaluated primary phakic trabeculectomies with no MMC (36 eyes of 36 patients), 0.5- to 1-minute MMC (50 eyes of 50 patients), and 3- to 5 minute MMC (38 eyes of 38 patients) at the concentration of 0.5 mg/ml. Successful trabeculectomy was defined as an intraocular pressure of 21 mm Hg or less without development of a marked visual acuity loss associated with prolonged hypotony (intraocular pressure < 6 mm Hg over 3 months) and without the need for additional surgery to control intraocular pressure or treat postoperative complications. RESULTS: The three groups were similar in demographics, preoperative intraocular pressure, and medical dependency. However, the incidence of hypotony during the postoperative periods of 3 to 12 months was significantly higher in the 3- to 5-minute MMC group (P < .05, chi-square test). Severe visual acuity loss associated with hypotony was also more frequently found in the 3- to 5-minute MMC group than in the 0.5- to 1-minute (P = .009, chi-square test) group or the control group (P = .014, chi-square test). In addition, the success probabilities were significantly different among the three groups (P = .001, Kaplan-Meier survival analysis with log-rank test) and were the highest in the 0.5- to 1-minute MMC group and the lowest in the 3- to 5-minute MMC group. CONCLUSION: Shorter application (0.5 to 1 minute) of MMC appears to be optimal for the successful outcome of primary phakic trabeculectomy compared with no MMC or longer application of MMC at a concentration of 0.5 mg/ml. PMID- 9859999 TI - Patterns of optic disk damage in patients with early focal visual field loss. AB - PURPOSE: To study the patterns of structural damage of the optic disk in patients with early focal visual field loss using a confocal scanning laser ophthalmoscope. METHODS: Thirty-nine subjects with repeatable early focal visual field loss were included. The Heidelberg Retina Tomograph (Heidelberg Engineering, Heidelberg, Germany) was used to obtain topographic optic disk measurements. For analysis of structural damage, the topographic measurements were divided into 36 10-degree sectors. Sector analysis was performed using sector rim area to sector disk area ratio. Patients were assigned to one of three groups based on the measurement of the optic disk damage: (1) diffuse disk damage; (2) focal disk damage; or (3) no detectable disk damage. RESULTS: Approximately half of the patients with early focal glaucomatous visual field loss showed diffuse optic disk damage, one quarter to one third had focal damage, and one sixth had no detectable damage. Optic disk area was smallest in the no detectable-damage group (1.71 + 0.19 mm2), followed by the focal-damage group (2.06 + 0.54 mm2), and was largest in the diffuse damage group (2.29 + 0.48 mm2; P = .22). CONCLUSION: In patients with early focal glaucomatous visual field loss, observable optic disk damage can be diffuse, focal, or undetectable. The Heidelberg Retina Tomograph may be capable of detecting different patterns of diffuse or focal structural damage. PMID- 9860000 TI - Refractive changes after pediatric intraocular lens implantation. AB - PURPOSE: To report refractive changes after cataract surgery and intraocular lens implantation in infants and children. METHODS: In an ongoing prospective study, the refractive errors of all patients younger than 18 years undergoing intraocular lens implantation were determined at 6 weeks, 3 months, 6 months, and 1 year, and at least yearly thereafter. All patients with greater than 6 months of follow-up were included in the study. RESULTS: Eighty-three eyes of 81 patients were identified. Cataracts were traumatic in 32 eyes (38%) and developmental in 42 eyes (50%). At implantation, the mean (+/-SD) age was 6.3 +/- 4.6 years (range, 9 months to 17 years). The mean follow-up was 26.6 months (range, 6 months to 6.6 years). Patients 0 to 2 years old at the time of implantation demonstrated a mean myopic shift of -3.00 diopters during a mean follow-up period of 2.5 years. Patients 2 to 6 years old at the time of implantation demonstrated a mean myopic shift of -1.50 diopters in a similar follow-up period. Children aged 6 to 8 years experienced a mean myopic shift of 1.80 diopters during a mean follow-up period of 3.0 years, while children older than 8 years at the time of intraocular lens implantation experienced a mean myopic shift of -0.38 diopters during a mean follow-up period of 1.8 years. On average, the operated-on eye showed a greater mean myopic shift than the fellow eye. No statistically significant differences in refractive change were found in comparing amblyopic to nonamblyopic eyes, traumatic to nontraumatic cataracts, or primary to secondary intraocular lenses. CONCLUSIONS: Our data demonstrate a trend toward increasing postoperative myopia in pediatric patients undergoing intraocular lens implantation. This myopic shift is greatest in the younger age groups and persists until at least 8 years of age. There is much variability in the postoperative refractive changes, and predicting exactly when and where the refraction will stabilize for an individual patient is difficult. PMID- 9860002 TI - Reperfusion of occluded capillary beds in diabetic retinopathy. AB - PURPOSE: To demonstrate the reperfusion of nonperfused capillary beds in diabetic retinopathy. METHODS: In a retrospective study, we reviewed 292 fluorescein angiograms of 94 eyes of 74 patients (mean age, 52 years; range, 20 to 68 years) with diabetic retinopathy. Fluorescein angiography was performed repeatedly (mean, three times; range, two to eight times) during a mean follow-up period of 2 years (range, 3 months to 12 years). None of the 94 eyes received laser photocoagulation. RESULTS: Reperfusion of occluded capillary beds was observed in 65 (69%) of 94 eyes. Reperfusion was characterized by recanalization in 22 (34%) of the 65 eyes or by intraretinal neovascularization in 54 (83%) of the 65 eyes. The former took place in small nonperfused areas and the latter in larger nonperfused areas. Reperfusion occurred throughout the entire fundus in six of 94 eyes, resulting in resolution of diabetic retinopathy. Reperfused capillary beds with intraretinal neovascularization left vascular remodeling, which was seen as twisted or kinked abnormal vessels. CONCLUSIONS: In diabetic retinopathy, occluded capillary beds may be reperfused. Twisted abnormal vessels may represent the reperfusion process through intraretinal neovascularization. PMID- 9860001 TI - Diabetes, cardiovascular disease, selected cardiovascular disease risk factors, and the 5-year incidence of age-related cataract and progression of lens opacities: the Beaver Dam Eye Study. AB - PURPOSE: To describe the relationships of diabetes mellitus, cardiovascular disease, and selected cardiovascular disease risk factors to cumulative incidence of age-related cataract and to progression of lens opacities over a 5-year interval. METHODS: A follow-up examination of the Beaver Dam Eye Study cohort was performed 5 years after the baseline evaluation. Ages at the census prior to baseline ranged from 43 to 84 years of age. Protocols for examination, lens photography, and grading were the same for both examinations. RESULTS: Age at baseline was the most significant characteristic associated with incidence of nuclear, cortical, and posterior subcapsular cataract in those without diabetes (P < .001) for all cataracts. The positive association of age with cataract was found for nuclear and cortical cataract in the worse eye (P < or = .04) but not posterior subcapsular cataract in those with diabetes. Progression of nuclear sclerosis was common, occurring in about 70% of subjects when considering either eye. Incident cortical and posterior subcapsular cataracts (P < or = .001 for worse eye for each lesion) and progression of cortical and posterior subcapsular opacities were more common in those with diabetes (P < or = .001 for either eye for each lesion). Increased glycated hemoglobin level was associated with increased risk of nuclear and cortical cataracts in those with diabetes. Relationships of risk factors to posterior subcapsular cataracts, especially among those with diabetes, were often in the expected direction but lacked significance possibly due to small samples. CONCLUSIONS: Diabetes mellitus is associated with incidence over 5 years of cortical and posterior subcapsular cataract and with progression of more minor cortical and posterior subcapsular lens opacities. These changes may be related to level of glycemia. Cardiovascular disease and its risk factors have little effect on incidence of any age-related cataract. PMID- 9860004 TI - Human leukocyte antigens associated with hyperthyroid Graves ophthalmology in Japanese patients. AB - PURPOSE: To evaluate the association of human leukocyte antigen (HLA) types in Japanese patients with Graves ophthalmopathy. METHODS: Japanese patients with severe ophthalmopathy (48 patients) and patients without ophthalmopathy (46 patients) were selected prospectively from 234 consecutive patients with hyperthyroid Graves disease. The criteria for severe ophthalmopathy were proptosis of 19 mm or more, or a 3-mm difference in exophthalmometry measurement between the two eyes, or diplopia at the primary eye position. The criteria for the absence of ophthalmopathy were proptosis of 16 mm or less, no diplopia, no lid retraction, and no thickening of extraocular muscles. Serologic HLA typing of class I and II antigens was performed in the two patient groups. The frequency of each HLA antigen in the two patient groups was compared to that in 767 unrelated normal healthy Japanese controls. RESULTS: The frequencies of HLA-DR14 and DQ1 were significantly higher in the patient group with severe ophthalmopathy (35% and 75%, respectively) than in healthy controls (4% and 43%, respectively; corrected P < .05), and also greater than in the patient group without ophthalmopathy (13% and 55%, respectively; P < .01). On the other hand, the frequencies of HLA-B35, B54, DR4, and DQ4 were significantly higher in the patient group without ophthalmopathy (23%, 23%, 43%, and 30%, respectively) than in the patient group with severe ophthalmopathy (13%, 15%, 25%, and 19%, respectively; P < .01). CONCLUSIONS: HLA-DR14 and DQ1 antigens may be genetic markers of predisposition to the development of severe ophthalmopathy. In addition, HLA-B35, B54, DR4, and DQ4 may be genetic markers of resistance to severe ophthalmopathy. Variation in the severity of ophthalmopathy associated with Graves disease may depend, in part, on the presence or absence of particular HLA antigens. PMID- 9860003 TI - The effect of silicone ocular surgical devices on serum IgG binding to silicones. AB - PURPOSE: To determine whether silicone materials used in retinal detachment repair and cataract surgery increase serum IgG binding to silicone and identify correlations with complications of ocular surgery. METHODS: Serum from 49 patients who had ocular surgery using silicone materials was examined. Patient groups included scleral buckling (n = 25), silicone oil tamponade (n = 3), scleral buckling and silicone oil tamponade (n = 9), and silicone lens implants after cataract extraction (n = 12). Convalescent samples for all patients and preoperative samples from 19 patients (18 scleral buckling and one silicone oil tamponade) were examined. Postoperative complications were monitored for up to 108 months (mean, 10.7 months; mode, 1.5 months; range, 1 to 108 months). Samples were evaluated for the extent of IgG binding to silicones using a micromodification of a previously described enzyme-linked immunosorbent assay method. RESULTS: In 19 patients, IgG binding levels in preoperative samples were 21 arbitrary units (AU) or less. Of the 25 buckling patients, one developed complications; however, in all patients the postoperative levels of IgG binding to silicone were low (2.2 to 20.0 AU). Although four silicone lens patients developed mild complications, none displayed postoperative IgG binding levels of greater than 20 AU. Three patients who underwent both scleral buckling and silicone oil tamponade developed complications; one of these patients, who was also noted to have systemic connective tissue disease, had a significant elevation in postoperative serum IgG binding to silicone. CONCLUSIONS: Statistically significant elevations of serum IgG binding to silicone were noted postoperatively in only one patient who had a systemic connective tissue disease. The complication rate and frequency of enhanced serum IgG binding to silicone was low, making correlations to surgical complications difficult. Examination of matched samples suggested that if ocular exposure to silicone implants enhances the level of serum IgG binding to silicones, it must be a rare event that should not alter the clinical use of these important devices. PMID- 9860005 TI - Balloon catheter dilation for treatment of adults with partial nasolacrimal duct obstruction: a preliminary report. AB - PURPOSE: The purpose of this study is to determine the efficacy and morbidity of balloon catheter dilation for treatment of partial acquired nasolacrimal duct obstruction in adults with epiphora. METHODS: We performed balloon dacryocystoplasty prospectively on a series of 15 partial nasolacrimal duct obstructions in 13 adults with epiphora. Partial obstruction was diagnosed by a negative Jones-1 test and canalicular irrigation revealing simultaneous reflux through the opposing punctum and drainage into the nose. Balloon dacryocystoplasty was performed under local anesthesia using an antegrade insertion technique. Silicone intubation of the nasolacrimal system was performed immediately after balloon catheter dilation, and the tubes were removed 2 months postoperatively. RESULTS: Success was measured objectively and subjectively at follow-up examinations 2 months and 6 months after the procedure. At 2 months, 11 (73%) of 15 obstructions were open on irrigation, with subjective success (Munk, grade 0 or grade 1) reported in 13 (87%) of 15 obstructions. At 6 months, 11 (73%) of 15 obstructions were open on irrigation, with subjective success (Munk, grade 0 or grade 1) reported in 9 (60%) of 15 obstructions. CONCLUSIONS: Balloon dacryocystoplasty may be a satisfactory primary treatment for adults with acquired partial nasolacrimal duct obstruction who exhibit no clinical signs of chronic infection. Additional long-term observations following balloon dacryocystoplasty are required. PMID- 9860006 TI - Discontinuing anticytomegalovirus therapy in patients with immune reconstitution after combination antiretroviral therapy. AB - PURPOSE: To describe our experience with discontinuation of anticytomegalovirus maintenance therapy in patients who have had immune reconstitution after initiation of highly active antiretroviral therapy. METHODS: Fifteen patients with treated cytomegalovirus retinitis, who had immune reconstitution after initiation of highly active retroviral therapy, had anticytomegalovirus maintenance therapy discontinued. Patients were followed closely for relapse of retinitis. RESULTS: Median nadir CD4+ T-cell count, before institution of highly active antiretroviral therapy, was 20 cells/microl. At the time of discontinuation of anticytomegalovirus therapy, median CD4+ T-cell count was 297 cells/microl. Patients were followed for a median of 8 months off anticytomegalovirus therapy (range, 3 to 16 months). The median CD4+ T-cell count at last follow-up was 267 cells/microl. No patient off anticytomegalovirus therapy relapsed. CONCLUSION: In selected patients with immune reconstitution after initiation of highly active antiretroviral therapy, anticytomegalovirus therapy may be safely discontinued, at least temporarily. Longer follow-up of these patients is needed to determine how long such therapy may be interrupted, and when anticytomegalovirus therapy should be reinstituted. PMID- 9860007 TI - Mitomycin C trabeculectomy in eyes with cicatricial conjunctiva. AB - PURPOSE: To demonstrate favorable outcome of mitomycin C-augmented trabeculectomy in eyes with broad cicatricial conjunctiva created by previous surgeries. METHODS: Forty-six eyes (40 patients) with extensive conjunctival scarring that had undergone mitomycin C trabeculectomy were reviewed retrospectively. RESULTS: After a mean follow-up +/- SD of 13.7 +/- 7.8 months (range, 6 to 36 months), intraocular pressure was well controlled, below or equal to 16 mm Hg and 21 mm Hg, respectively, in 33 (72%) and 44 (96%) of the 46 eyes. In all eyes, a functional filtering bleb was present during the follow-up periods. CONCLUSION: Mitomycin C trabeculectomy after dissection of conjunctival scar tissue may be useful for treating refractory glaucoma. PMID- 9860008 TI - Lasso procedure to revise overcorrection with radial keratotomy. AB - PURPOSE: To report three patients who underwent the lasso procedure to revise overcorrection with radial keratotomy. METHODS: Case report and review of the literature. RESULTS: Four eyes of three patients who had undergone radial keratotomy with resultant hyperopic overcorrection underwent a lasso procedure. Before the procedure, average cycloplegic refraction spherical equivalent was +3.656 +/- 1.352 diopters, and average manifest refraction spherical equivalent was +2.250 +/- 0.621 diopters. A 10.0 monofilament nylon suture was placed in a circumferencial manner through the corneal stroma and overlapping the old radial keratotomy incisions. At 1 month postoperatively, best-corrected visual acuity was 20/20 in all four eyes, with average cycloplegic refraction spherical equivalent +0.438 +/- 1.423 diopters and average manifest refraction spherical equivalent -0.156 +/- 1.147 diopters. Mean delta cycloplegic refraction spherical equivalent was 3.219 +/- 1.724. CONCLUSIONS: The lasso procedure provides an immediate solution for symptomatic overcorrected hyperopic eyes after radial keratotomy. Predictability and long-term stability necessitate further follow-up. PMID- 9860009 TI - Transepithelial photorefractive keratectomy for treatment of thin flaps or caps after complicated laser in situ keratomileusis. AB - PURPOSE: To report transepithelial photorefractive keratectomy treatment of corneal irregularities produced during laser in situ keratomileusis (LASIK) in which there is a thin flap or cap associated with central corneal scarring or epithelial ingrowth that threatens vision. METHODS: Case reports. The thickness of the abnormal corneal flap or cap and associated scarring or epithelial ingrowth is estimated at the slit lamp or measured with an optical pachymeter. If residual myopia is sufficiently high to allow complete ablation of the flap or cap in the central cornea, a transepithelial photorefractive keratectomy is performed in which the epithelium is completely ablated with the excimer laser in phototherapeutic keratectomy mode; residual myopia is corrected using photorefractive keratectomy. RESULTS: This method was used successfully in two eyes of two patients in which a thin cap was associated with a transverse cut through the central cornea or a donut-shaped flap associated with epithelial ingrowth in the central cornea. In both cases, the abnormal cap or flap was ablated, central corneal clarity restored, and visual function improved. CONCLUSION: Transepithelial photorefractive keratectomy may be effective in treating central corneal thin cap or flap abnormalities associated with LASIK. PMID- 9860010 TI - Photorefractive keratectomy for myopia in the setting of adenoviral subepithelial infiltrates. AB - PURPOSE: To report a patient with persistent subepithelial corneal opacities 18 months after adenoviral keratoconjunctivitis who underwent photorefractive keratectomy for the correction of myopia. METHODS: Case report, review of medical literature, and slit-lamp photography. RESULTS: The patient underwent photorefractive keratectomy in each eye, 1 week apart, with ablation of central corneal opacities and resultant best-corrected visual acuity of BE, 20/20. Symptomatic subepithelial stromal infiltrates recurred in the peripheral but not the central cornea of each eye 3 months after laser treatment. CONCLUSIONS: Photorefractive keratectomy was successful in the correction of myopia and prevented the recurrence of adenoviral subepithelial corneal infiltrates within the laser-treated central cornea. PMID- 9860011 TI - Gelatinous drop-like corneal dystrophy is not one of the beta ig-h3-mutated corneal amyloidoses. AB - PURPOSE: To discover if beta ig-h3 is mutated in gelatinous drop-like corneal dystrophy, as has been suggested. METHODS: Genomic DNA was isolated from unrelated individuals with lattice corneal dystrophy type I (n = 3), Avellino corneal dystrophy (n = 3), and gelatinous drop-like corneal dystrophy (n = 3) and used as a template for polymerase chain reaction to amplify all exons in beta ig h3. The polymerase chain reaction product was then sequenced. RESULTS: Beta ig-h3 is mutated in lattice corneal dystrophy type I (Arg124Cys) and Avellino corneal dystrophy (Arg124His). In gelatinous drop-like corneal dystrophy, on the other hand, no mutation was detected in the entire coding region of beta ig-h3 (all 17 exons). CONCLUSION: Unlike the amyloidotic corneal dystrophies lattice type I and Avellino, gelatinous drop-like corneal dystrophy is not likely to be caused by a mutation in beta ig-h3. PMID- 9860012 TI - Corneal opacities in Gaucher disease. AB - PURPOSE: To describe the corneal findings in a variant of Gaucher disease. METHODS: Case report. In an 18-year-old man, ophthalmic and general clinical evaluation, and enzymatic and molecular genetics studies were performed. RESULTS: Diffuse, well-defined, small, linear, or dotlike corneal opacities were observed through, out the posterior two thirds of the corneal stroma in both eyes. The patient had calcific valvular heart disease. Enzymatic and ultrastructural studies were consistent with Gaucher disease. Analysis of the glucocerebrosidase gene disclosed homozygosity for a D409H mutation. CONCLUSION: Corneal opacities are a distinguishing ocular feature of the variant of Gaucher disease associated with the D409H mutation and with calcific cardiac disease. PMID- 9860013 TI - The survival of adenovirus in multidose bottles of topical fluorescein. AB - PURPOSE: To determine if common ocular adenovirus serotypes survive in vitro in multidose bottles of topical fluorescein (Fluress; Pilkington Barnes Hind, Inc, Sunnyvale, California). METHODS: Clinical isolates of adenovirus types 8 and 19 were inoculated separately into 10 bottles each of Fluress and maintained at room temperature (25 C). All bottles were titered for adenovirus on A549 cell monolayers at 0, 7, 14, 21, 28, and 49 days. RESULTS: Adenovirus was recovered from Fluress for up to 21 days for adenovirus type 19 and 28 days for adenovirus type 8. CONCLUSION: A multidose bottle of Fluress contaminated with adenovirus can be a potential source of adenoviral transmission in an ophthalmic office setting. PMID- 9860014 TI - Rash, fever, and chills after intravenous fluorescein angiography. AB - PURPOSE: To report a previously unreported complication associated with intravenous injection of fluorescein dye. METHOD: Case report. A 75-year-old man developed a unique complication after intravenous injection of fluorescein dye for angiography. RESULTS: Two hours after receiving an intravenous injection of fluorescein for angiography, the patient developed a fever, rash, and chills. Admission to a hospital and careful systemic evaluation determined that this reaction was a noninfectious allergic response to intravenous fluorescein dye injection. CONCLUSION: A delayed allergic response to intravenous fluorescein dye injection can occur. PMID- 9860015 TI - Ultrasound biomicroscopic analysis of anterior proliferative vitreoretinopathy. AB - PURPOSE: To report the use of ultrasound biomicroscopy for preoperative assessment of anterior proliferative vitreoretinopathy. METHODS: Case report. In a 35-year-old man with tractional retinal detachment and anterior proliferative vitreoretinopathy, we used ultrasound biomicroscopy with UX-02 (Rion Co Ltd, Tokyo, Japan) to perform the preoperative analysis of the ciliary body, anterior vitreous, and detached peripheral retina. RESULTS: In all meridians, the posterior insertion of the vitreous was drawn anteriorly, creating a retinal trough, and the meridian distance between the anterior and posterior vitreous base insertions was reduced. Retinochoroidal detachment was detected posterior to the anterior vitreous base insertion. Based on these imagings, sclerotomy locations were selected. CONCLUSION: Ultrasound biomicroscopy may be useful to assess anterior proliferative vitreoretinopathy preoperatively and to decide on sclerotomy sites. PMID- 9860016 TI - Anatomic and functional recovery of the fovea after foveal translocation surgery without large retinotomy and simultaneous excision of a neovascular membrane. AB - PURPOSE: To document the anatomic and functional recovery of the fovea after foveal translocation surgery with scleral shortening and simultaneous excision of a neovascular membrane in a patient with age-related macular degeneration. METHOD: Case report. RESULTS: The visual acuity of a 54-year-old woman with age related macular degeneration improved from 20/200 to 20/50 after excision of subretinal neovascular membrane and foveal translocation surgery in the right eye. Fixation shifted inferonasally 0.6 disk diameters, corresponding to the direction of foveal translocation, as shown by scanning laser ophthalmoscope microperimetry. Postoperative optical coherence tomography through fixation disclosed normal foveal concavity and intact retinal pigment epithelium. CONCLUSION: Anatomic and functional recovery of the fovea was confirmed in a patient with age-related macular degeneration after foveal translocation surgery with scleral shortening and simultaneous excision of a neovascular membrane. PMID- 9860017 TI - Three-dimensional ultrasonography of choroidal melanoma: extrascleral extension. AB - PURPOSE: To describe the results of three-dimensional ultrasonography used to evaluate extrascleral extension of a choroidal melanoma. METHODS: Case report. The three-dimensional ultrasound system uses a 10-MHz B-mode transducer combined with a motorized rotating holder. The system acquires 180 sequential images that are stored and processed to create a three-dimensional block of the region of interest. RESULTS: Unique coronal and oblique perspectives were obtained from interactive manipulation of the three-dimensional reconstruction. Examination of the three-dimensional image allowed us to detect the transscleral uveal-orbital connection. Extrascleral melanomatous extension was confirmed on histopathologic examination. CONCLUSION: Three-dimensional ultrasonography is a promising imaging technique for evaluating melanomatous extrascleral extension. PMID- 9860018 TI - Epibulbar allergic granulomatous nodules in a human immunodeficiency virus positive patient. AB - PURPOSE: To report an unusual epibulbar inflammatory process in a patient with human immunodeficiency virus (HIV). METHODS: Case report. A 32-year-old man developed fleshy epibulbar nodules on his right conjunctiva and cornea after being treated for conjunctivitis. A biopsy of the lesions was done, and the specimen was processed for histopathologic examination. RESULTS: The biopsy specimen contained inflammatory cells, including an eosinophilic abscess. The diagnosis was allergic granulomatous nodules. CONCLUSION: This case illustrates the occurrence of epibulbar allergic granulomatous nodules in an HIV-positive patient. PMID- 9860019 TI - Trichoadenoma of the eyelid. AB - PURPOSE: To report the unique occurrence of a trichoadenoma of the eyelid. METHODS: An 80-year-old woman underwent surgical resection of a lesion of the right lower eyelid that was suspected to be a basal cell carcinoma. The lesion was studied with light microscopy. RESULTS: Histopathology showed a lesion in the dermis that contained keratin cysts surrounded by a proliferation of eosinophilic epidermoid cells. The findings were characteristic of a trichoadenoma. CONCLUSION: Trichoadenoma, a rare, benign, cutaneous tumor that usually occurs on the face, can develop on the eyelid and simulate a basal cell carcinoma. PMID- 9860020 TI - Orbital floor implant migration across the ethmoidal sinuses and nasal septum. AB - PURPOSE: To report an unusual case of orbital floor implant migration across the ethmoidal sinuses and nasal septum. METHOD: Case report. A 61-year-old woman with a history of right orbital floor fracture repair 25 years earlier is described. RESULTS: The patient presented with sinus congestion and difficulty breathing through the right nostril. Computed tomographic scan disclosed medial migration of the right orbital floor implant across the ethmoidal sinuses and nasal septum. The patient underwent transorbital and transnasal endoscopic surgery with removal of the implant. CONCLUSIONS: When an alloplastic orbital floor implant is required, size and fixation of the implant are important. Late paranasal sinus or nasal airway problems may be sequelae, and the possibility of implant migration should be considered. PMID- 9860021 TI - Ishihara color plates as a test for simultanagnosia. AB - PURPOSE: To alert ophthalmologists to the use of Ishihara color plates in the detection of simultanagnosia. METHODS: We examined seven patients referred for impaired vision. Evaluation included color plate testing with Ishihara color plates. RESULTS: All seven patients had simultanagnosia, with marked difficulty in identifying the numbers in Ishihara color plates despite adequate visual acuity and the ability to name all of the colors in the plates correctly. One of these patients was referred with the diagnosis of a cone dystrophy because of her poor performance on the Ishihara test. All of the patients had bilateral occipitoparietal damage or atrophy on magnetic resonance imaging. CONCLUSIONS: Ophthalmologists must be aware that a poor performance with Ishihara plates may not be attributable to an impairment of color vision but rather to occipitoparietal brain damage associated with simultanagnosia. PMID- 9860022 TI - The ophthalmology of intracranial abnormalities. PMID- 9860023 TI - Unilateral peripapillary myelinated retinal nerve fibers associated with strabismus, amblyopia, and myopia. PMID- 9860024 TI - Executive Summary of Joint Task Force Practice Parameters on Diagnosis and Management of Rhinitis. AB - Rhinitis is a significant cause of widespread morbidity, medical treatment costs, reduced work productivity and lost school days. Although sometimes mistakenly viewed as a trivial disease, symptoms of allergic and non-allergic rhinitis may significantly impact a patient's quality of life, by causing fatigue, headache, cognitive impairment and other systemic symptoms. In addition, many antihistamines commonly used for treatment can themselves cause performance impairment that may contribute to fatal automobile accidents, work place accidents, decreased work productivity and in children, impaired school performance. Appropriate management of rhinitis may be an important component in effective management of coexisting or complicating respiratory conditions, such as asthma, sinusitis, or chronic otitis media. Rhinitis may be caused by allergic, non-allergic, infectious, hormonal, occupational, and other factors. Defining the causes of rhinitis in an individual is important because different rhinitis syndromes may require different therapeutic approaches for optimal management, an important consideration as more treatment options become available. This Executive Summary reviews key points about diagnosis and management of rhinitis contained in the comprehensive document, Diagnosis and Management of Rhinitis: Complete Guidelines of Joint Task Force on Practice Parameters in Allergy, Asthma and Immunology, and Joint Task Force Algorithm and Annotations for Diagnosis and Management of Rhinitis. These documents represent a consensus opinion of the Joint Task Force on Practice Parameters in Allergy, Asthma and Immunology, a national panel co-sponsored by the American Academy of Allergy, Asthma and Immunology, the American College of Allergy, Asthma and Immunology, and the Joint Council on Allergy, Asthma and Immunology. PMID- 9860025 TI - Joint Task Force Algorithm and Annotations for Diagnosis and Management of Rhinitis. AB - The algorithm and text annotations in this document are intended to assist clinical decision making about patients who present with symptoms of rhinitis. This document complements the Executive Summary of Joint Task Force Practice Parameters for Diagnosis and Management of Rhinitis (Ann Allergy, Asthma, Immunol 1998; 81:463-468) and Diagnosis and Management of Rhinitis: Complete Guidelines of the Joint Task Force on Practice Parameters in Allergy, Asthma and Immunology (Ann Allergy, Asthma, Immunol 1998;81:478-578). The Joint Task Force on Practice Parameters in Allergy, Asthma and Immunology is co-sponsored by the American Academy of Allergy, Asthma and Immunology, the American College of Allergy, Asthma and Immunology and the Joint Council of Allergy, Asthma and Immunology. PMID- 9860026 TI - Joint Task Force summary statements on Diagnosis and Management of Rhinitis. PMID- 9860027 TI - Diagnosis and management of rhinitis: complete guidelines of the Joint Task Force on Practice Parameters in Allergy, Asthma and Immunology. American Academy of Allergy, Asthma, and Immunology. AB - This document contains complete guidelines for diagnosis and management of rhinitis developed by the Joint Task Force on Practice Parameters in Allergy, Asthma and Immunology, representing the American Academy of Allergy, Asthma and Immunology, the American College of Allergy, Asthma and Immunology and the Joint Council on Allergy, Asthma and Immunology. The guidelines are comprehensive and begin with statements on clinical characteristics and diagnosis of different forms of rhinitis (allergic, non-allergic, occupational rhinitis, hormonal rhinitis [pregnancy and hypothyroidism], drug-induced rhinitis, rhinitis from food ingestion), and other conditions that may be confused with rhinitis. Recommendations on patient evaluation discuss appropriate use of history, physical examination, and diagnostic testing, as well as unproven or inappropriate techniques that should not be used. Parameters on management include use of environmental control measures, pharmacologic therapy including recently introduced therapies and allergen immunotherapy. Because of the risks to patients and society from sedation and performance impairment caused by first generation antihistamines, second generation antihistamines that reduce or eliminate these side effects should usually be considered before first generation antihistamines for the treatment of allergic rhinitis. The document emphasizes the importance of rhinitis management for comorbid conditions (asthma, sinusitis, otitis media). Guidelines are also presented on special considerations in patients subsets (children, the elderly, pregnancy, athletes and patients with rhinitis medicamentosa); and when consultation with an allergist-immunologist should be considered. PMID- 9860028 TI - What's in a name? Vaccine versus extracts. PMID- 9860029 TI - Pathophysiology of nocturnal asthma. AB - LEARNING OBJECTIVES: This article will focus on the pathophysiologic changes underlying the nocturnal worsening of asthma and the therapeutic approach to this disorder. DATA SOURCES: Selected articles appearing since 1985 dealing specifically with the underlying pathologic features and therapy of nocturnal asthma. STUDY SELECTION: Studies that aimed to elucidate the pathologic features, mechanisms, and therapeutic strategies for the treatment of nocturnal asthma are summarized. RESULTS: Nocturnal asthma is associated with significant decline in pulmonary function and increase of airway inflammation at night. The administration of medications must be designed to achieve the maximal effect during the night in nocturnal asthma. CONCLUSIONS: The further elucidation of the reasons underlying nocturnal asthma should lead to more specific therapeutic interventions with maximal effect at night. PMID- 9860030 TI - Latex antigens: identification and use in clinical and experimental studies, including crossreactivity with food and pollen allergens. AB - LEARNING OBJECTIVES: The purpose of this review is to introduce the reader to the range of latex allergens that have been identified by polypeptide sequencing. This knowledge is important for the assessment of clinical latex hypersensitivity, including crossreactivity with food and aeroallergens. DATA SOURCES: Medline search and relevant publications and reviews from the English medical literature since 1989. RESULTS: Of the more than 150 polypeptides in natural latex rubber, 35 or more can act as allergens and are recognized by IgE antibodies in the sera of latex-sensitive subjects. Complete or partial amino acid sequence data have now been obtained for 20 or more allergens, and have facilitated cloning of genes and development of allergen-specific antibodies. In latex-sensitive adults, hevein (Hev b6), rubber elongation factor (Hev b1) and Hev b5 are reported as major allergens, while in children with spina bifida, latex particle proteins are important allergens. Although incomplete, the data for finished latex products indicate that the number as well as diversity of latex antigens are limited compared with the allergen profile of natural rubber latex. Latex allergen sequence data allows epitope mapping, which establishes the molecular basis for understanding crossreactivity between latex, food, and aeroallergens. CONCLUSION: The identification of latex allergens is paving the way for studying the molecular basis of immunoreactivity to these proteins, including crossreactivity with food and other allergens. The development of reagents to detect and measure antigen-specific responses will increase diagnostic specificity of allergic reactions to latex products. PMID- 9860032 TI - Inhaled corticosteroids-the FDA takes another look. PMID- 9860031 TI - Allergen immunotherapy: therapeutic vaccines for allergic diseases. World Health Organization. American academy of Allergy, Asthma and Immunology. PMID- 9860033 TI - Algorithm for the diagnosis and management of asthma: a practice parameter update: Joint Task Force on Practice Parameters, representing the American Academy of Allergy, Asthma and Immunology, the American College of Allergy, Asthma and Immunology, and the Joint Council of Allergy, Asthma and Immunology. AB - This algorithm on the diagnosis and treatment of asthma is intended to complement and update the previously published Practice Parameters for the Diagnosis and Treatment of Asthma. Both documents were developed by the Joint Task Force on Practice Parameters, representing the AAAAI, ACAAI, and the JCAAI. The authors of this asthma algorithm have attempted to include all the elements essential for the diagnosis and care of patients with asthma. Every effort was made to keep the algorithm clear and concise, yet thorough and complete (Fig 1). Each component of the algorithm is elaborated further in a brief annotation. For further discussion, the reader is referred to the more extensive Practice Parameters for the Diagnosis and Treatment of Asthma. PMID- 9860035 TI - Fluorescein-induced allergic reaction. AB - BACKGROUND: Adverse reactions following intravenous sodium fluorescein are very unusual and their mechanism is still uncertain. We report the case of a patient who suffered an adverse reaction during a fluorescein ocular angiography. Positive allergy tests to fluorescein suggest an IgE-mediated mechanism. OBJECTIVE: Report the allergy evaluation performed in a patient who suffered an adverse reaction during an intravenous fluorescein administration. METHODS: We selected the case of a patient who suffered dizziness, diaphoresis, generalized pallor, nausea, sphincter relaxation, hypotension, and intense malaise during a fluorescein ocular angiography and compared the results to other nonreactive subjects. Allergy evaluation: Prick and intradermal skin tests and serial determinations of serum tryptase were performed on the patient and four control subjects who underwent and tolerated the same procedure as well as on a patient who developed an intense vagal reaction during blood extraction. RESULTS: Positive skin tests and dramatic increase of serum tryptase (67U/I) were observed in our patient. The rest of the patients had negative skin tests and did not have any variation in their serum tryptase. CONCLUSIONS: An IgE-mediated mechanism is suggested as responsible for this adverse reaction. We recommend that a complete allergy evaluation should be performed in all patients who have adverse reactions to fluorescein in order to differentiate true allergic reactions from other types of reactions. PMID- 9860034 TI - A case of pathophysiologic study in Kimura's disease: measurement of cytokines and surface analysis of eosinophils. AB - BACKGROUND: Kimura's disease is a rare but distinctive eosinophilic inflammatory disorder of unknown etiology; few reported case studies have focused on the immunopathologic background of this unique disease. OBJECTIVE: To define better the immunopathogenetic features of Kimura's disease, we attempted to quantitatively analyze values of cytokines and soluble interleukin-2 receptor (sIL-2R) in peripheral blood (PB), as well as perform surface immunophenotypic analysis of eosinophils from a Japanese patient with chronic relapsing Kimura's disease. RESULTS: Granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-alpha) and sIL-2R were elevated, and newly expressed antigens on eosinophils CD4, CD25, and HLA-DR were found to be involved in the pathophysiology of this disorder. CONCLUSIONS: Kimura's disease may be a disease in which activated lymphocytes release cytokines, and these released cytokines, such as GM-CSF and TNF-alpha cause eosinophil activation. These processes may be related to the pathogenesis of this disorder. PMID- 9860036 TI - Effect of mometasone furoate on early and late phase inflammation in patients with seasonal allergic rhinitis. AB - BACKGROUND: Mometasone furoate is a potent glucocorticoid that can markedly inhibit proinflammatory Th2 cytokines in vitro. An aqueous nasal spray formulation has been shown to be clinically active in reducing the symptoms of perennial and seasonal allergic rhinitis. OBJECTIVE: To determine whether pretreatment with mometasone furoate 200 microg once daily decreases specific indices of early and late phase nasal inflammation compared with placebo. METHODS: A randomized, double-blind, placebo-controlled crossover study was conducted using nasal provocation with ragweed antigen in 21 patients with ragweed-induced allergic rhinitis out of the ragweed season; the treatment period was 2 weeks. Symptom scores, rhinoprobe cytology, and nasal lavage fluid were collected during early and late phase periods for nasal cytokines (interleukin, 1, 4, 5, 6, and 8) and leukotriene B4 determinations using ELISA and RIA. RESULTS: Mean nasal symptom scores and sneezing frequency were consistently lower with mometasone furoate compared with placebo. Treatment was associated with a statistically significant early phase (30-minute time point) reduction in nasal lavage histamine levels compared with placebo (14.3 versus 20.2 ng/mL, P = .02). Within-treatment comparisons suggested that mometasone furoate reduced the antigen-induced late-phase response for IL-6, IL-8, and eosinophils compared with pretreatment. There were similar, but smaller, changes seen in the placebo group for these measurements. There were no statistically significant changes following antigen challenge in IL-1, IL-4, IL-5, LTB4, or in other nasal cytology parameters. CONCLUSION: These results suggest that the clinical activity of mometasone furoate nasal spray in seasonal allergic rhinitis is likely due, in part, to a reduction in the levels of histamine in nasal secretions related to the early phase response, and reductions in IL-6, IL-8, and eosinophils during the late phase response. PMID- 9860037 TI - A survey of television meteorologists about their sources for and understanding of pollen counts. AB - BACKGROUND: Pollen counts are widely used by television stations in news programming but little is known about the sources for and quality of this information. OBJECTIVE: The purpose of this investigation was to survey television meteorologists about their sources for pollen counts and to assess their general understanding of these data. METHODS: Sixty-seven meteorologists employed by television stations in the nation's 20 largest Designated Market Areas (DMAs) were contacted by telephone. A brief survey was administered verbally and their responses were recorded. RESULTS: Thirty-seven meteorologists indicated that their station used pollen counts in news programming. Most were aware of their station's source for this information and reported that viewers contacted them about the pollen count. Few meteorologists, however, knew the type of instrument that was used to obtain pollen samples they aired and only 10 selected the correct definition of a pollen count from a list of three choices. CONCLUSIONS: Pollen counts are an important element in television news programming that are poorly understood by the meteorologists who present them to the public. Allergists are encouraged to better educate the personnel at stations to which they supply pollen counts. PMID- 9860038 TI - Interleukin 4 and interferon-gamma secretion by antigen and mitogen-stimulated mononuclear cells in the hyper-IgE syndrome: no TH-2 cytokine pattern. AB - BACKGROUND: Enhanced production of TH-2 cytokines plays a key role in increased IgE production in allergic diseases. Reports about the cytokine profile secreted by peripheral blood mononuclear cells of patients with hyper-IgE syndrome, however, are controversial, suggesting alternative causes for increased IgE production in this syndrome. OBJECTIVE: We wished to determine whether mononuclear cells from patients with hyper-IgE syndrome have a pattern of interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) production characteristic of a predominance of TH-2 cells and whether the cytokine production pattern is constant over time. METHODS: IL-4 and IFN-gamma secretion by peripheral blood mononuclear cells stimulated with phytohemagglutinin and D. pteronyssinus was measured by ELISA in culture supernatants. Patients with the hyper-IgE syndrome were evaluated 3 times at 4-week intervals and compared with asthmatic patients and normal subjects. RESULTS: In PHA-stimulated cultures, patients with hyper-IgE syndrome had an IL-4 and IFN-gamma secretion similar to that of controls, while asthmatic patients had increased IL-4 and decreased IFN-gamma production. Cultures stimulated with D. pteronyssinus showed a variable pattern of secretion for both cytokines. CONCLUSIONS: In allergic diseases, increased serum IgE level is the result of a TH-2 pattern of cytokine production, with high IL-4 and decreased IFN-gamma protein secretion. The increased serum IgE concentration typical of the hyper-IgE syndrome is likely the result of a different immunoregulatory process. PMID- 9860039 TI - Inhibitory effect of ibudilast (KC-404) on the expression of the beta2 integrin family on an eosinophilic cell line (EoL-1). AB - BACKGROUND: Adhesion molecules are thought to play a key role in inflammatory processes in bronchial asthma. We previously observed an increased expression of the beta2-integrin family on an eosinophilic cell line (EoL-1) by platelet activating factor (PAF). OBJECTIVE: In the present study, we examined the effect of ibudilast (KC-404), a novel anti-asthma agent, on beta2 integrin expression induced by PAF. MATERIAL AND METHODS: EoL-1 cells (1x10(6)/mL) were incubated in the presence or absence of 10(-6) M ibudilast (KC-404), then cells were cultured in the presence or absence of PAF (10(-7) M) for 45 minutes. Flow cytometric analysis for CD11a, CD11b, and CD18 expression was examined. RESULTS: Ibudilast had an inhibitory effect on beta2 integrin expression induced by PAF [CD11a: 84.8% versus 73.1% (preincubation with ibudilast), CD11b: 35.8% versus 26.2%, CD18 74.9% versus 65.6%]. CONCLUSIONS: Ibudilast (KC-404) has anti-inflammatory activities through its inhibitory effect on the expression of adhesion molecules on eosinophils. PMID- 9860040 TI - Topical sodium cromoglycate in the treatment of moderate-to-severe atopic dermatitis. AB - BACKGROUND: Unsatisfactory treatment results for severe atopic dermatitis have led to many experimental therapies, including cromolyn sodium in various vehicles at concentrations ranging from 1% to 10%. Results suggest that the vehicle used to deliver the cromolyn is relevant to its effectiveness. OBJECTIVE: To test the efficacy of low concentrations of cromolyn in a water-soluble vehicle in the treatment of moderate-to-severe atopic dermatitis in a double-blind, placebo controlled study. METHODS: Twenty-six pediatric patients who had failed to respond to conventional therapy were randomized into 2 treatment groups: patients in group A used the study drug for 1 month (phase I), then received the placebo for 1 month (phase II); and patients in group B used the placebo for 1 month, then received the study drug for 1 month. The study drug was cromolyn sodium inhalation solution mixed into a water-based emollient cream to a final concentration of 0.21%. Upon enrollment and at each follow-up visit, every patient was given a severity score based on extent and severity of skin involvement. RESULTS: At enrollment, there were no significant differences between groups A and B in severity scores, age, sex, race, skin test and/or RAST positivity, eosinophil levels, IgE concentrations, or the presence of concomitant rhinitis or asthma. After the first phase of the study treatment, severity scores had decreased significantly for both groups with a significant difference between group A (cromolyn) and group B (placebo). After crossover, both groups had significantly lower severity scores than at entry into the study. CONCLUSION: Treatment with topical cromolyn in a hydrophilic emollient vehicle has a significant anti-inflammatory effect on moderate-to-severe atopic dermatitis. We have now incorporated this treatment into our clinical practice. PMID- 9860041 TI - Aspirin-induced isolated periorbital angioedema. PMID- 9860042 TI - The association between the total antioxidant potential of plasma and the presence of coronary heart disease and renal dysfunction in patients with NIDDM. AB - Oxidative stress may be an important pathogenetic factor in the development of diabetic vascular complications. The total antioxidative potential of plasma reflects the ability of an individual to resist oxidative stress. We measured the plasma total peroxyl radical-trapping potential (TRAP) and the concentrations of four plasma chain-breaking antioxidants in 81 patients with non-insulin-dependent diabetes mellitus (NIDDM) nine years after diagnosis and in 102 well-matched non diabetic control subjects. The association between the total antioxidative potential and the presence of coronary heart disease (CHD) and diabetic kidney disease were also studied. There were no significant differences in plasma TRAP between NIDDM patients and control subjects (1250+/-199 vs. 1224+/-198 microM). Nor were there any significant differences in the concentrations of plasma uric acid, ascorbic acid, alpha-tocopherol, and protein thiols between NIDDM patients and control subjects. Patients with a low glomerular filtration rate and/or high urinary albumin excretion had elevated plasma uric acid. Plasma TRAP was not, however, associated with renal dysfunction. The plasma of NIDDM patients with CHD had a significantly higher value of unidentified antioxidative potential than that of patients without CHD. This relation was strongly dependent upon smoking. In conclusion, these data demonstrate that there are no major defects in the antioxidative potential of plasma caused by NIDDM per se. CHD and diabetic renal dysfunction were not associated with changes in plasma TRAP. PMID- 9860043 TI - Pyruvate prevents hydrogen peroxide-induced apoptosis. AB - Studies were carried out to investigate the protective effects of pyruvate, a key glycolytic intermediate and alpha-keto-monocarboxylate, against oxidative stress induced apoptosis. Oxidative stress was induced by treating mouse thymocytes with 25 microM hydrogen peroxide for 15 min at 37 degrees C under 5% CO2 in air. Pre- and post-treatment of cells with 10 mM pyruvate inhibited morphological changes, internucleosomal DNA fragmentation, and translocation of phosphatidylserine to the plasma membrane surface, which are characteristic features of apoptosis. L lactate (10 mM) and acetate (10 mM) were ineffective in inhibiting apoptosis and appeared to be toxic to the cells under similar conditions. The results suggest that pyruvate has therapeutic potential for use in the treatment of oxidative stress-induced disorders associated with increased apoptosis. PMID- 9860044 TI - Comparison of protein oxidation and aldehyde formation during oxidative stress in isolated mitochondria. AB - Oxidative stress is known to cause oxidative protein modification and the generation of reactive aldehydes derived from lipid peroxidation. Extent and kinetics of both processes were investigated during oxidative damage of isolated rat liver mitochondria treated with iron/ascorbate. The monofunctional aldehydes 4-hydroxynonenal (4-HNE), n-hexanal, n-pentanal, n-nonanal, n-heptanal, 2 octenal, 4-hydroxydecenal as well as thiobarbituric acid reactive substances (TBARS) were detected. The kinetics of aldehyde generation showed a lag-phase preceding an exponential increase. In contrast, oxidative protein modification, assessed as 2,4-dinitrophenylhydrazine (DNPH) reactive protein-bound carbonyls, continuously increased without detectable lag-phase. Western blot analysis confirmed these findings but did not allow the identification of individual proteins preferentially oxidized. Protein modification by 4-HNE, determined by immunoblotting, was in parallel to the formation of this aldehyde determined by HPLC. These results suggest that protein oxidation occurs during the time of functional decline of mitochondria, i.e. in the lag-phase of lipid peroxidation. This protein modification seems not to be caused by 4-HNE. PMID- 9860045 TI - Protective effects of antioxidants against UVA-induced DNA damage in human skin fibroblasts in culture. AB - Ultraviolet A radiation (UVA, 320-400 nm) is mutagenic and induces genomic damage to skin cells. N-acetyl-cysteine (NAC), selenium and zinc have been shown to have antioxidant properties and to exhibit protective effects against UVA cytotoxicity. The present work attempts to delineate the effect of these compounds on genomic integrity of human skin fibroblasts exposed to UVA radiation using the single cell gel electrophoresis (SCGE) or Comet assay. The cells were incubated with NAC (5 mM), sodium selenite (0.6 microM) or zinc chloride (100 microM). Then cells were embedded in low melting point agarose, and immediately submitted to UVA fluences ranging from 1 to 6J/cm2. In the Comet assay, the tail moment increased by 45% (1 J/cm2) to 89% (6J/cm2) in non-supplemented cells (p)<0.01). DNA damage was significantly prevented by NAC, Se and Zn, with a similar efficiency from 1 to 4J/cm2 (p < 0.05). For the highest UVA dose (6J/cm2), Se and Zn were more effective than NAC (p < 0.01). PMID- 9860046 TI - Fructose induced deactivation of glucose-6-phosphate dehydrogenase activity and its prevention by pyruvate: implications in cataract prevention. AB - Glucose-6-phosphate dehydrogenase (G6PDH) is an important lens enzyme diverting about 14% of the tissue glucose to the hexose monophosphate shunt pathway. The main function of such a pronounced activity of the enzyme is to support reductive biosyntheses, as well as to maintain a reducing environment in the tissue so as to prevent oxy-radical induced damage and consequent cataract formation. Sugars are one of the well-known cataractogenic agents. Several reports suggest that the cataractogenic effect of the sugars in diabetes as well as in normal aging is initiated by the glycation of the proteins including the enzymes and subsequent formation of more complex and biologically inactive or harmful structures. In a diabetic lens the concentration of fructose exceeds significantly the concentration of glucose, suggesting that the contribution of fructosylation may be greater than that of glucosylation. These studies were undertaken to examine further the possibility that in addition to glycation, generation of oxygen free radicals by fructose and consequent oxidative modifications in certain enzymes may be an important participant in the cataractogenic process. This hypothesis was tested by using G6PDH. The enzyme was incubated with various levels of fructose (0-20mM) and its activity determined as a function of time. This led to a significant loss of its activity, which was prevented by superoxide dismutase, catalase, mannitol and myoinositol. Most interestingly, pyruvate at levels between 0.2 and 1.0 mM also offered substantial protection. Hence, the results, while elucidating further the mechanism of enzyme deactivation by sugars such as fructose, also demonstrate the possibility of therapeutic prevention of cataracts by pyruvate and other such keto acids, in diabetes and other disabilities involving oxygen free radicals in the pathogenetic process. PMID- 9860047 TI - Determination of oxidative DNA base damage by gas chromatography-mass spectrometry. Effect of derivatization conditions on artifactual formation of certain base oxidation products. AB - GC-MS is a widely used tool to measure oxidative DNA damage because of its ability to identify a wide range of base modification products. However, it has been suggested that the derivatization procedures required to form volatile products prior to GC-MS analysis can sometimes produce artifactual formation of certain base oxidation products, although these studies did not replicate previously-used reaction conditions, e.g. they failed to remove air from the derivatization vials. A systematic examination of this problem revealed that levels of 8-hydroxyguanine, 8-hydroxyadenine, 5-hydroxycytosine and 5 (hydroxymethyluracil) in commercial calf thymus DNA determined by GC-MS are elevated by increasing the temperature at which derivatization is performed in our laboratory. In particular, 8-hydroxyguanine levels after silylation at 140 degrees C were raised 8-fold compared to derivatization at 23 degrees C. Experiments on the derivatization of each undamaged base revealed that the artifactual oxidation of guanine, adenine, cytosine and thymine respectively was responsible. Formation of the above products was potentiated by not purging with nitrogen prior to derivatization. Increasing the temperature to 140 degrees C or allowing air to be present during derivatization did not significantly increase levels of the other oxidized bases measured. This work suggests that artifactual oxidation during derivatization is restricted to certain products (8 hydroxyguanine, 8-hydroxyadenine, 5-hydroxycytosine and 5-[hydroxymethyluracil]) and can be decreased by reducing the temperature of the derivatization reaction to 23 degrees C and excluding as much air possible. Despite some recent reports, we were easily able to detect formamidopyrimidines in acid-hydrolyzed DNA. Artifacts of derivatization are less marked than has been claimed in some papers and may vary between laboratories, depending on the experimental procedures used, in particular the efficiency of exclusion of O2 during the derivatization process. PMID- 9860048 TI - Inhibition of xanthine oxidase by pterins. AB - The effect of a panel of pterins on xanthine oxidase was investigated by measuring formation of urate from xanthine as well as formazan production from nitroblue tetrazolium. The pterin derivatives, depending on their chemical structure, decreased urate as well as formazan generation: 200 microM neopterin and biopterin suppressed urate formation (90% from baseline) and formazan production (80% from baseline) as well. Their reduced forms, 7,8-dihydroneopterin and 5,6,7,8-tetrahydrobiopterin, showed a lesser but still strongly diminishing influence (40% from baseline). Another oxidized pterin namely leukopterin showed only a weak inhibitory effect. Xanthopterin, a known substrate of xanthine oxidase, had a strong effect on urate formation (80% inhibition), but a lesser effect on formazan production (30% reduction). When iron-(III)-EDTA complex was added to the reaction mixture all the effects were more pronounced. Superoxide dismutase, which removes superoxide anion by dismutation into oxygen, decreased formazan production in addition to pterin derivatives and had a small but enhancing effect on urate formation. Also the reductant N-acetylcysteine had an additive effect to pterins to diminish formazan production in a dose-dependent way. The results of our study suggest that depending on their chemical structure pterins reduce superoxide anion generation by xanthine oxidase. PMID- 9860049 TI - Two-electron electrochemical oxidation of quercetin and kaempferol changes only the flavonoid C-ring. AB - Bulk electrolysis of the antioxidant flavonoids quercetin and kaempferol in acetonitrile both yield a single oxidation product in two-electron processes. The oxidation products are more polar than their parent compounds, with an increased molecular weight of 16g/mol, and were identified as 2-(3,4-dihydroxybenzoyl) 2,4,6-trihydroxy-3(2H)-benzofuranone and 2-(4-hydroxybenzoyl)-2,4,6-trihydroxy 3(2H)-benzofuranone for quercetin and kaempferol, respectively. Two-electron oxidation of the parent flavonoid is suggested to yield a 3,4-flavandione with unchanged substitution pattern in the A- and B-ring, which may rearrange to form the substituted 3(2H)-benzofuranone through the chalcan-trione ring-chain tautomer. The acidity of the 3-OH group is suggested to determine the fate of the flavonoid phenoxyl radical, originally formed by one-electron oxidation, as no well-defined oxidation product of luteolin (lacking the 3-OH group) could be isolated despite rather similar half-peak potentials: Ep/2 = 0.97V, 0.98 V and 1.17 V vs. NHE for quercetin, kaempferol and luteolin, respectively, as measured by cyclic voltammetry in acetonitrile. PMID- 9860050 TI - Antioxidant properties of catechins and proanthocyanidins: effect of polymerisation, galloylation and glycosylation. AB - A range of catechins and oligomeric procyanidins was purified by high performance liquid chromatography (HPLC) from grape seed, apple skin, lentil and almond flesh. Catechins, galloylated epicatechin, glycosylated catechin, procyanidin dimers, galloylated dimers, trimer, and tetramer species were all identified, purified and quantified by HPLC, LC-MS and NMR. The antioxidant properties of these compounds were assessed using two methods: (a) inhibition of ascorbate/iron induced peroxidation of phosphatidylcholine liposomes; (b) scavenging of the radical cation of 2,2'-azinobis(3-ethyl-benzothiazoline-6-sulphonate) (ABTS) relative to the water-soluble vitamin E analogue Trolox C (expressed as Trolox C equivalent antioxidant capacity, TEAC). Antioxidant activity in the lipid phase decreased with polymerisation in contrast with antioxidant action in the aqueous phase which increased from monomer to trimer and then decreased from trimer to tetramer. Galloylation of catechin and dimeric procyanidins decreased lipid phase and increased aqueous phase antioxidant activity. Glycosylation of catechin demonstrated decreased activity in both phases. PMID- 9860051 TI - Oral administration of (-)catechin protects against ischemia-reperfusion-induced neuronal death in the gerbil. AB - The effect of ad libitum oral-administration of (-)catechin solution on ischemia reperfusion-induced cell death of hippocampal CA1 in the gerbil was histologically examined. When (-)catechin solution instead of drinking water was orally administered ad libitum for 2 weeks, dose-dependent protection against neuronal death following by transient ischemia and reperfusion was observed. To evaluate the involvement of reduction of reactive-oxygen-species (ROIs) by the antioxidant activity of (-)catechin in this protection, the superoxide scavenging activity of the brain in catechin-treated gerbils was measured by ESR and spin trapping using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). The superoxide scavenging activities of the brains obtained from catechin-treated gerbils were significantly higher than those of catechin-untreated animals. From these results, it was suggested that orally administered (-)catechin was absorbed, passed through the blood-brain barrier and that delayed neuronal death of hippocampal CA1 after ischemia-reperfusion was prevented due to its antioxidant activities. PMID- 9860052 TI - Review of studies that compare the quality of cardiovascular care in HMO versus non-HMO settings. AB - OBJECTIVES: The authors compared the quality of cardiovascular care in health maintenance organizations (HMOs) versus traditional insurance arrangements through an analysis of existing literature. METHODS: Data were derived from all peer-reviewed studies published through November 1995 that used process or outcome measures to evaluate the quality of cardiovascular care in HMO versus non HMO settings. A standardized form was used to extract information from each study on: condition studied, study time frame, type of study design, type of comparison groups, characteristics of patients and physicians, process and outcome measures used, data collection methods, reliability and validity of quality measurements, risk adjustment techniques, findings about quality of care, summary of other findings, study limitations, and other comments that explained the context of the research. RESULTS: Seven of the 11 studies that examined process measures for cardiovascular care in HMO versus non-HMO patients found more differences in one or more process measures that favored HMOs than non-HMOs. Seven of the 10 studies that examined outcome measures found no statistically significant differences in patient care between HMO and non-HMO settings. The other three studies presented contradictory results. CONCLUSIONS: The existing literature suggests that the outcomes of care for cardiovascular conditions do not differ between HMO and non HMO settings, although selected measures of the process of cardiovascular care are actually better in HMO than in non-HMO settings. PMID- 9860053 TI - The effect of staff nursing on length of stay and mortality. AB - OBJECTIVES: Specialized hospital units developed historically for the efficiency of physicians, but their existence has created an opportunity for staff nurses to specialize as well. This study was done to test the hypothesis that specialized staff nursing has an effect on patient outcome as length of stay (LOS) and mortality, using casemix information and controlling for physician volume. METHODS: Sixteen Diagnosis Related Groups associated with particular specialty units in Yale New Haven Hospital were selected. Five years of data (FY 1987-FY 1993) from a period in which specialized unit configuration was relatively stable were obtained (N = 11,316). Data elements included basic patient characteristics, especially diagnosis and procedure codes, physician identifiers as scrambled code numbers, length of stay, length of intensive care unit stay, and discharge disposition. Specialized nursing units were defined by the percentage of patients in a given diagnosis related group discharged from that unit. Patient age and differential intensive care unit use were used for risk adjustment. RESULTS: In 13 of the 16 diagnosis related groups, patients cared for on specialized nursing units had shorter lengths of stay; the difference was statistically significant in nine. In the seven Diagnosis Related Groups with any deaths, the mortality on the specialized unit(s) was lower; the difference was statistically significant in four. Physician volume, defined as more or fewer than 20 discharges per diagnosis related group had little or no effect on either length of stay or mortality. CONCLUSIONS: The notion that nurses improve at caring for similar patients of a stable group of physicians as their experience increases has common sense appeal. If the findings of this study can be replicated in other institutions, with the refinements suggested here, it may be possible to separate the effects of multidisciplinary practice on outcomes and to track the effect of hospital reengineering projects that change patient mix or nursing specialization. Studying one hospital in depth suggested that interhospital studies of cost and quality may need to consider nursing specialization along with other comparisons. PMID- 9860054 TI - An event count model for studying health services utilization. AB - OBJECTIVES: The purpose of this article is to first conceptualize health services utilization behavior as event counts. Based on this concept and behavioral theory, the author presents the Generalized Event Count model as an alternative modeling tool for studying health services utilization. This model is theory driven and is consistent with behavioral assumptions. METHODS: In presenting the Generalized Event Count model, the author first examines its model assumptions to see whether they conform to elements of behavior theory, assumptions of health services utilization, and the distribution assumption of the nature of the data. To demonstrate the Generalized Event Count model, the author applied this model to an empirical ambulatory care utilization data set from a 1988 household interview of Chinese-Americans in Boston's inner-city community. RESULTS: The Generalized Event Count model analysis suggested that the regular source of medical care and the use of Chinese medicine were strong predictors of physician visits for this population. Further, the Generalized Event Count model was able to test that most of the ambulatory visits within an individual were correlated. CONCLUSIONS: Compared with other models, the Generalized Event Count model is more consistent with health services utilization behavioral assumptions. Moreover, it makes an efficient use of information from the utilization data for model estimation. This model has the potential of having broad applications in studying various types of health services utilization, especially for analyzing cross-sectional utilization data. PMID- 9860055 TI - Interpreting risk-adjusted length of stay patterns for VA hospitals. AB - OBJECTIVES: The Veterans Health System must become more competitive with the private sector in terms of efficiency of care. Studies have shown significantly longer lengths-of-stay (LOS) in facilities operated by the Department of Veterans Affairs (VA) compared with private sector facilities. Most comparisons, however, have not controlled well for casemix differences or have involved small numbers of patients. The aims of this study were: (1) controlling for casemix, to accurately measure the degree by which average length of stay in Veterans Affairs facilities exceeds that of private sector hospitals and (2) to demonstrate a methodology with which individual VA facilities can identify clinical and demographic subgroups of patients associated with the higher length-of-stay averages. METHODS: Subjects of the study were Veterans Health System patients hospitalized during 1991-1993 and veteran respondents to the 1991 National Hospital Discharge Survey. Hospitals' mean length of stay adjusted for patients' diagnosis related groups, severity, demographics, and travel distances were measured. RESULTS: Veterans Affairs medical centers' average risk-adjusted length of stay was 36% higher (8.9 days compared with 6.5 days) than that of the private sector. For individual hospitals, relative length-of-stay efficiency typically varied by condition. Among 14 hospitals in the VA's midwest region, none were high risk-adjusted length-of-stay outliers in all conditions studied, and four were high outliers for some conditions and low outliers for others. CONCLUSIONS: Controlling for differences in patient demographic and clinical factors, Veterans Affairs medical centers consumed significantly more days of care than private sector hospitals. Veterans Affairs medical centers will be able to improve efficiency by identifying specific subgroups of patients whose clinical treatment should be examined. PMID- 9860056 TI - Comparison of health status, socioeconomic characteristics, and knowledge and use of HIV-related resources between HIV-infected women and men. AB - OBJECTIVES: The authors compared socioeconomic characteristics, and knowledge and use of human immunodeficiency virus (HIV)-related resources and health status measures between HIV-infected women and men registered within the Denver Health and Hospitals health care system. METHODS: Data collected through two Centers for Disease Control-funded surveillance initiatives (Adult Spectrum of Disease and Supplement to HIV/AIDS Surveillance) were linked. Health status measures were obtained using the Medical Outcomes Study (MOS-20) questionnaire. To compare health status measures between genders, men were matched to women based on disease stage, intravenous drug use, race, years of education, employment status, and age. RESULTS: Among all patients interviewed (n = 419), women (n = 52) were more likely to be minority, uneducated, intravenous drug users, and at earlier stages of HIV-disease than men (n = 367). Employment status was not significantly different. Knowledge of available services was generally good among both genders. Women received public assistance and had health insurance (Medicaid) more often than men. Women used support services, social work, and shelter assistance less often than men. The matched pairs analysis (n = 46 pairs) showed no significant differences between genders in physical and social function, mental health, pain, or general health perceptions; however, role function was better in women than in men (P<0.02). CONCLUSIONS: When controlling for factors that may influence health and access to health care, HIV disease generally impacts the health status of both genders similarly. Women scored higher in role function which may reflect family caretakers' responsibilities. Although knowledge of HIV-related resources was similar by gender, men made contact more often suggesting areas for enhanced outreach toward women. PMID- 9860057 TI - Uncoupling of blood flow and metabolism in focal epilepsy. AB - PURPOSE: Interictal measurements of cerebral blood flow are less helpful in localizing epileptic foci than are measurements of brain metabolism. This may be related to an uncoupling of blood flow and metabolism. In this study, brain metabolism and blood flow were compared in an acute experimental model of focal interictal epilepsy. METHODS: Interictal epileptic foci were induced by an epicortical application of penicillin in rats. After 1 h, stereotyped interictal activity was initiated, lasting until the end of the experiment. Brain metabolism was determined with [14C]deoxyglucose, and cerebral blood flow with [14C]iodoan tipyrine autoradiography. RESULTS: In control experiments, metabolism and blood flow were coupled. In animals with focal interictal epileptic activity, the metabolism was strongly increased in the focus and reduced in areas lateral to the focus. In contralateral brain areas, blood flow and metabolism varied in a parallel fashion. Ipsilateral to the focus, however, blood flow and metabolism were altered disproportionately. In the focus, the increase of blood flow was less marked than the increase of metabolism, and the area with increased blood flow was larger than the area with increased metabolism. Lateral to the focus, in the area with a hypometabolism, blood flow was not concomitantly reduced. CONCLUSIONS: The experiments show that blood flow and metabolism in focal epilepsy may be uncoupled in widespread regions. This is due neither to structural abnormalities nor to the duration or discharge pattern of epileptic activity. The results explain why interictal metabolic investigations have a higher predictive value in presurgical epilepsy evaluation than do interictal measurements of blood flow. PMID- 9860058 TI - Bicuculline-induced rhythmic EEG episodes: gender differences and the effects of ethosuximide and baclofen treatment. AB - PURPOSE: There are gender differences in the expression of seizures. We tested rhythmic EEG episodes induced by low doses of bicuculline in rats for gender differences. To verify the validity of these discharges as a model of absence seizures in both male and female rats, we tested the antiabsence drug ethosuximide (ESM) and a gamma-aminobutyric acid (GABA(B))-receptor agonist, baclofen, which may exacerbate absence seizures. METHODS: Adult rats of both sexes were used. Under general anesthesia, EEG electrodes were implanted over frontal and occipital cortex, and some females were ovariectomized. After recovery, male, intact female rats, and female rats ovariectomized and ovariectomized rats with estradiol replacement were compared for occurrence of rhythmic EEG episodes (approximately 6 cycles/ s) induced by 2.5 mg/kg of bicuculline, s.c. Because of gender differences in sensitivity to bicuculline, further pharmacologic effects of ESM (125 and 250 mg/kg, i.p.) and baclofen (2 mg/kg, i.p.) were tested separately in male (3.0 mg/kg of bicuculline), and female (2.5 mg/kg of bicuculline) rats. RESULTS: After the identical dose of bicuculline, s.c., male and female rats differed in the incidence of rhythmic episodes and in the latency to onset of the first as well as the generalized episode. Female rats with natural or exogenous estrogens (but not ovariectomized rats) developed EEG episodes more often than did males, and this effect could be attributed to the presence of estrogens. ESM pretreatment suppressed the episodes, whereas baclofen enhanced their occurrence, as well as the total duration of episodes without gender-specific differences. CONCLUSIONS: The study demonstrates gender differences (related probably to the presence of circulating estrogens) in the susceptibility of rats to develop rhythmic EEG episodes induced by threshold doses of bicuculline. This activity has some features of an acute absence seizure model. PMID- 9860059 TI - Evidence of enhanced kindling and hippocampal neuronal injury in immature rats with neuronal migration disorders. AB - PURPOSE: Neuronal migration disorders (NMD) are often found in patients with epilepsy. However, the mechanisms linking these two pathologies are not yet fully understood. In this study, we evaluated whether NMD increased kindling seizure susceptibility and seizure-induced acute neuronal damage in the immature brain. METHODS: Experimental NMD were produced by exposing pregnant rats (gestation day 15) to methylazoxymethanol acetate (MAM, 25 mg/kg, ip). Seizures were induced in rat pups (postnatal day 15) transplacentally exposed to MAM and controls by hippocampal kindling. Afterdischarge (AD) threshold and duration, seizure stage, and number of stimulations required to reach each seizure stage were recorded. Acute seizure-induced damage was histologically assessed in Nissl-stained and silver-impregnated hippocampal tissue 24 h after kindling. RESULTS: Rat pups with NMD had a significantly lower AD threshold than controls (91+/-18 vs. 163+/-23 microA; p < 0.05). Furthermore, rats with NMD required fewer stimulations to reach seizure stage 3.5 and 4 than did controls. Additionally, rats with NMD had longer AD the second day of stimulation (2,094+/-416 s vs. 1,755+/-353 s; p < 0.05). Histologic examination revealed that in rats with NMD, acute seizure induced neuronal hippocampal damage occurred bilaterally in CA3 hippocampal neurons. CONCLUSIONS: The lowered AD threshold and more rapid kindling to stages 3.5 and 4 indicate that in the presence of severe NMD, hippocampal kindling is facilitated. Furthermore, this study suggests that in the immature brain, seizure induced hippocampal neuronal damage occurs if there is an underlying pre-existing pathology. PMID- 9860060 TI - Activation of the locus coeruleus after amygdaloid kindling. AB - PURPOSE: Substantia nigra (SN) and locus coeruleus (LC) neurons are implicated in the propagation and suppression of amygdaloid seizures. Both structures are activated concomitant with amygdaloid seizure discharges. Their mechanisms of activation, however, remain to be elucidated. SN firing is not associated with the induction of Fos immunoreactivity (ir), a marker of excitatory neuronal activation. LC has not been studied. The purpose of this investigation was to determine if amygdala-kindled generalized seizures could induce Fos-ir in the LC. METHODS: Female Sprague-Dawley rats were killed after generalized seizures induced by amygdala electrical stimulation and stained by using Fos immunocytochemistry. The number of Fos-ir neurons was compared between 15 animals with generalized seizures and four implanted, unstimulated controls. RESULTS: LC ir neurons were significantly (p < 0.05) more prevalent after seizures than in control animals. Their numbers correlated very highly with Fos-ir in the central nucleus of the amygdala (p < 0.0001). No Fos induction was observed in LC in controls or in the SN in either group. CONCLUSIONS: Amygdala-induced generalized seizures result in Fos-ir in the LC but not in the SN. This is consistent with different mechanisms of activation possibly involving disinhibition in the SN and direct excitation in the LC. PMID- 9860061 TI - Typical absence status in adults: diagnostic and syndromic considerations. AB - PURPOSE: To study the electroclinical features of typical absence status (TAS) in adults with syndromes of idiopathic generalized epilepsies (IGEs). METHODS: Twenty-one patients with one or more spells of TAS were identified among 136 consecutive adult patients with IGEs. All patients with TAS had comprehensive electroclinical investigations and EEG or video-EEG recorded absences. RESULTS: TAS occurred in 24.4% of 86 patients who had IGEs with typical absences alone or in combination with other seizures presisting in adult life. The prevalence of TAS appeared to be syndrome related, ranging from as high as 57.1% in perioral myoclonia with absences and 46.2% in "phantom" absences with GTCS to as low as 6.7% in juvenile myoclonic epilepsy. A varying degree of impairment of cognition was the cardinal clinical symptom shared in all TAS, but corresponding syndromes of IGE were often betrayed by other symptoms such as eyelid or perioral myoclonia. In phantom absences with GTCS, TAS was more numerous (p < or = 0.05) and more frequently the first overt seizure type (p = 0.006) than in any other IGE. Only in the syndrome of eyelid myoclonia with absences, TAS was always situation related, mainly as a result of antiepileptic drug discontinuation. CONCLUSIONS: The clinical EEG semiology and prevalence of TAS appear to be syndrome related with the highest prevalence in the syndromes of perioral myoclonia with absences and phantom absences with GTCS (p = 0.0024). PMID- 9860062 TI - Analysis of ictal EEGs of epilepsy associated with tuberous sclerosis. AB - PURPOSE: To investigate the significance of cortical pathology of tonic spasms in patients with tuberous sclerosis. METHODS: The subjects were 38 patients with epilepsy associated with tuberous sclerosis. We analyzed ictal EEGs of tonic spasms and partial seizures by means of video-EEG monitoring for a total of 763 tonic spasms in 20 patients and 107 partial seizures in 15 patients. We also investigated the relation between partial seizures and magnetic resonance imaging (MRI) findings of these patients. RESULTS: Ictal EEG patterns of tonic spasms were divided into generalized and focal patterns. Thirteen patients had only generalized patterns, whereas seven had both patterns. In five patients who had focal ictal patterns of tonic spasms and partial seizures, the location of focal patterns corresponded with the location of onset of partial seizures. Focal discharges were seen immediately before, after, and in the middle of tonic spasms in series in 13 patients. The location of focal discharges also corresponded with the location of the onset of partial seizures in 10 of the 13 patients. Regarding partial seizures, four patients had multiple active epileptogenic foci during the same period, and two others had shifting epileptogenic foci with increasing age. CONCLUSIONS: These findings indicate that cortical pathology plays an important role in the occurrence not only of partial seizures but also of tonic spasms in patients with tuberous sclerosis. PMID- 9860063 TI - Cerebral hemodynamic response to generalized spike-wave discharges. AB - PURPOSE: Data in the literature concerning metabolic demand during generalized spike-wave activity (gSW) are conflicting. We investigated instantaneous changes in cerebral blood flow velocities (CBFV) in both middle cerebral arteries (MCAs) by transcranial Doppler sonography (TCD) during gSW paroxysms recorded by scalp EEG. METHODS: In 13 patients, CBFVs in both MCAs were averaged, time-locked to the occurrence of the gSW; respiratory rate (RR) and end-expiratory pco2 were measured in one patient. RESULTS: Nine patients showed significant changes in CBFV during gSW. Four had biphasic flow changes with an initial increase (p < 0.05) and a subsequent decrease (p < 0.01). This was partially paralleled by an increase in RR (p < 0.01) and a decrease in pco2 (p < 0.01). In three patients, an increase in CBFV that preceded the onset of gSW by several seconds was observed, followed by a decrease in CBFV. Two patients showed a significant decrease only of CBFV. Only gSWs of a median duration of >0.8 s were associated with significant changes in CBFV. CONCLUSIONS: We were able to demonstrate that gSWs of several seconds duration lead to cortical perfusion changes. We suggest that the initial increase of CBFV demonstrated in some patients reflects neuronal activation, whereas the subsequent decrease might in part be due to hyperventilation-induced hypocapnia. PMID- 9860064 TI - Comparison of combined versus subdural or intracerebral electrodes alone in presurgical focus localization. AB - PURPOSE: The yield of subdural versus intracerebral electrodes for ictal localization remains a point of controversy. We assessed the relative sensitivity of these two types of electrodes per case. METHODS: Eighty-three intracranial recordings obtained from 82 patients were retrospectively reviewed to establish which type of electrode performed best in which patients and which seizure types. RESULTS: Sixty (73%) of 82 patients had temporal lobe seizure onsets, eight frontal, nine widespread or multifocal/multilobar or both, whereas in five, seizure onset was not localized. Exclusive use of intracerebral electrodes would have been sufficient for accurate localization of the seizure-onset zone in all 35 patients with strictly mesial temporal seizure onsets. In only 20 (57%) of these 35 patients, the same decision would have been reached with exclusive use of subdural electrodes. In widespread neocortical and mesial temporal seizures (n = 25), yield of both electrode types was at about the same level, but neither was sufficient to identify the zone of ictal onset on its own. In frontal or multilobar seizures (n = 22), yield of subdural electrodes was slightly better then that of the intracerebral electrodes, but was not sufficient in all cases. CONCLUSIONS: This study indicates that, depending on the characteristics of the seizure disorder, exclusive use of either intracerebral or subdural electrodes may easily result in erroneous diagnosis because of insufficient sampling of the brain. These findings are in contrast with other studies emphasizing the high yield of reliable EEG findings in evaluations with a single type of electrode and corroborate the results of one of our previous studies. PMID- 9860065 TI - Chronic intracranial EEG monitoring for localizing the epileptogenic zone: an electroclinical correlation. AB - PURPOSE: To evaluate the diagnostic yield and identify predictive factors of the surgical outcome in patients with intractable partial epilepsy undergoing chronic intracranial EEG monitoring (CIEM). METHODS: The clinical, magnetic resonance imaging (MRI) and electrophysiologic data of 108 patients that underwent CIEM were retrospectively reviewed. The discharge pattern and spatial extent of the initial ictal discharge were determined by blinded visual inspection and computerized analysis. RESULTS: The main predictive indicator for epilepsy surgery outcome in patients that underwent CIEM was the presurgical MRI findings. Most patients with hippocampal atrophy or complete lesionectomy were rendered seizure free after epilepsy surgery (83 and 80%, respectively), whereas only a small minority of patients with partial lesionectomy or no detected MRI lesion had seizure-free operative outcomes (21 and 22%, respectively). Multifocal independent initiation of the initial ictal discharge was associated with a poor surgical outcome. In contrast, the pattern and local spatial extent of the initial ictal discharge observed with CIEM failed to predict the surgical outcome. CONCLUSIONS: The main predictor of the surgical outcome in patients that underwent CIEM was the MRI findings, whereas CIEM had only limited use in localizing the epileptogenic zone in the absence of an MRI lesion. The reported findings indicate a low specificity of CIEM in defining the site of seizure onset, which in turn significantly impairs the reliability of CIEM in delineating the epileptogenic zone for epilepsy surgery. Further studies are required to define the indications and patient subpopulations who can benefit from CIEM before epilepsy surgery. PMID- 9860066 TI - Interictal spiking increases with sleep depth in temporal lobe epilepsy. AB - PURPOSE: To test the hypothesis that deepening sleep activates focal interictal epileptiform discharges (IEDs), we performed EEG-polysomnography in 21 subjects with medically refractory temporal lobe epilepsy. METHODS: At the time of study, subjects were seizure-free for > or =24 h and were taking stable doses of antiepileptic medications (AEDs). Sleep depth was measured by log delta power (LDP). Visual sleep scoring and visual detection of IEDs also were performed. Logistic-regression analyses of IED occurrence in relation to LDP were carried out for two groups of subjects, nine with frequent IEDs (group 1) and 12 with rare IEDs (group 2). RESULTS: The LDP differentiated visually scored non-rapid eye movement (NREM) sleep stages (p = 0.0001). The IEDs were most frequent in NREM stages 3/4 and least frequent in REM sleep. Within NREM sleep, in both groups, IEDs were more frequent at higher levels of LDP (p < 0.05). In group 1, after accounting for the level of LDP, IEDs were more frequent (a) on the ascending limb of LDP and with more rapid increases in LDP (p = 0.007), (b) in NREM than in REM sleep (p = 0.002), and (c) closer to sleep onset (p < 0.0001). Fewer than 1% of IEDs occurred within 10 s of an EEG arousal. CONCLUSIONS: Processes underlying the deepening of NREM sleep, including progressive hyperpolarization in thalamocortical projection neurons, may contribute to IED activation in partial epilepsy. Time from sleep onset and NREM versus REM sleep also influence IED occurrence. PMID- 9860067 TI - The effects of genetic polymorphisms of CYP2C9 and CYP2C19 on phenytoin metabolism in Japanese adult patients with epilepsy: studies in stereoselective hydroxylation and population pharmacokinetics. AB - PURPOSE: The aim of this study was to clarify the effects of genetic polymorphisms of cytochrome P450 (CYP) 2C9 and 2C19 on the metabolism of phenytoin (PHT). In addition, a population pharmacokinetic analysis was performed. METHODS: The genotype of CYP2C9 (Arg144/Cys, Ile359/Leu) and CYP2C19(*1, *2 or *3) in 134 Japanese adult patients with epilepsy treated with PHT were determined, and their serum concentrations of 5-(4-hydroxyphenyl)-5 phenylhydantoin (p-HPPH) enantiomers, being major metabolites of PHT, were measured. A population pharmacokinetic analysis (NONMEM analysis) was performed to evaluate whether genetic polymorphism of CYP2C9/19 affects the clinical use of PHT by using the 336 dose-serum concentration data. RESULTS: The mean maximal elimination rate (Vmax) was 42% lower in the heterozygote for Leu359 allele in CYP2C9, and the mean Michaelis-Menten constants (Km) in the heterozygous extensive metabolizers and the poor metabolizers of CYP2C19 were 22 and 54%, respectively, higher than those without the mutations in CYP2C9/19 genes. (R)- and (S)-p-HPPH/PHT ratios were lower in patients with mutations in CYP2C9 or CYP2C19 gene than those in patients without mutations. CONCLUSIONS: Although the hydroxylation capacity of PHT was impaired with mutations of CYP2C9/19, the impairment was greater for CYP2C9. In view of the clinical use of PHT, two important conclusions were derived from this population study. First, the serum PHT concentration in patients with the Leu359 allele in CYP2C9 would increase dramatically even at lower daily doses. Second, the patients with CYP2C19 mutations should be treated carefully at higher daily doses of PHT. PMID- 9860068 TI - A pilot study of topiramate in the treatment of infantile spasms. AB - PURPOSE: West syndrome is a rare epileptic syndrome associated with infantile spasms, a specific abnormal electroencephalographic pattern (termed hypsarrhythmia), and mental retardation. Management of this disorder is difficult because current treatment regimens, including many anticonvulsants and hormones, are often ineffective. Topiramate (TPM) is a new antiepileptic drug that may be effective in pediatric epilepsies. We conducted a pilot study to test the effects of rapid TPM dosing in patients with refractory infantile spasms. METHODS: Eleven children with refractory infantile spasms were given an initial dose of 25 mg TPM per day in addition to their current therapy. Dosage was increased by 25 mg every 2-3 days until spasms were controlled, the maximal tolerated dose was reached, or the maximal dose of 24 mg/kg/day was achieved. Efficacy was primarily assessed by video EEG and secondarily by parental count of spasm frequency. RESULTS: Five (45%) subjects became spasm free during the study, with absence of infantile spasms and hypsarrhythmia (either classic or modified) proven by video EEG. Nine subjects, including the five spasm free, achieved a spasm reduction of > or =50%. Spasm frequency decreased from 25.6+/-19.3 to 6.9+/-5.9 spasms/day. Sixty-four percent of the subjects were able to achieve TPM monotherapy. CONCLUSIONS: Results in this cohort of 11 patients with refractory disease show TPM to be a promising new agent for the treatment of infantile spasms. PMID- 9860069 TI - Double-blind, placebo-controlled, crossover study of lamotrigine in treatment resistant generalised epilepsy. AB - PURPOSE: Lamotrigine (LTG) is recognised as effective add-on therapy for focal epilepsies, but this is the first double-blind, placebo-controlled, crossover study in treatment-resistant generalised epilepsy. METHODS: The study consisted of 2 x 8-week treatment periods followed by a 4-week washout period. Patients received doses of either 75 or 150 mg daily, depending on their concomitant antiepileptic drugs (AEDs). Long-term continuation was offered at the end of the study with open-label LTG. RESULTS: Five centres in Australia recruited 26 patients who were having absence, myoclonic, or generalized tonic-clonic seizures or a combination of these. Twenty-two patients completed the study. There was a significant reduction in frequency of both tonic-clonic and absence seizure types with LTG. A 350% decrease in seizures was observed for tonic-clonic seizures in 50% of cases and for absence seizures in 33% of evaluable cases. Rash was the only adverse effect causing discontinuation. Twenty-three of 26 opted for open label LTG, with 20 still receiving LTG for a mean of 26 months. In these 20, 80% had > or =50% seizure reduction and five (25%) were seizure free. CONCLUSIONS: This study shows that LTG is effective add-on therapy in patients with refractory generalised epilepsies. Statistically significant reduction in seizures in both absence and tonic-clonic seizure types was seen even with low doses of LTG. PMID- 9860070 TI - Neurocysticercosis in persons with epilepsy in Medellin, Colombia. The Neuroepidemiological Research Group of Antioquia. AB - PURPOSE: A prospective series of 643 persons with epilepsy attending a reference neurologic center in Medellin, Colombia, was examined by computed tomography (CT scan) or serology or both with the enzyme-linked immunoelectrotransfer blot assay (EITB) to assess the prevalence of Taenia solium cysticercosis. METHODS: All presenting patients were consecutively enrolled in the study. Five hundred forty six persons underwent cerebral CT scans; 376 of them also had serum EITB performed. RESULTS: Prevalence of neurocysticercosis by CT scan was 13.92%. Overall prevalence of T. solium antibodies with EITB was 9.82%, but for those with late-onset epilepsy (onset after age 30 years), prevalence increased to 17.5% and 19% for those who originated from outside urban Medellin. Seroprevalence in individuals with mixed lesions (cysts and calcifications) was 88.2% and 64.10% in those with live cysts. Conversely, only 2.72% of persons with CT findings not related to neurocysticercosis had positive EITB tests. CONCLUSIONS: Our study shows that an important proportion of individuals with epilepsy have radiologic or serologic evidence of T. solium infection, suggesting that neurocysticercosis is an important etiology for epilepsy in Colombia. PMID- 9860071 TI - Photosensitive seizures provoked while viewing "pocket monsters," a made-for television animation program in Japan. AB - PURPOSE: To describe the recent epidemic of photosensitive seizure that occurred in relation to an episode of the television animation program "Pocket Monsters," we report four patients who experienced seizures while watching the episode in question. We also report some technical aspects of the program episode. METHODS: We investigated the clinical symptoms of the four patients and performed routine EEGs with intermittent photic stimulation (IPS). If IPS provoked no photoparoxysmal response (PPR) during the routine EEG examination, a second EEG was performed with the photic stimulator placed 10 cm from the patient's eyes. In addition, we reviewed the "Pocket Monsters" episode, focusing our attention on the visual techniques used with reference to the Independent Television Commission (ITC) guidelines. RESULTS: One patient who had myoclonic jerks before the convulsion in question was diagnosed as having juvenile myoclonic epilepsy, and the diagnosis of another patient was pure photosensitive epilepsy. The remaining two patients had their first seizures, which could be occasional seizures, and we therefore could not reach a diagnosis of epilepsy. In our four patients, only one showed PPR on the routine EEG. Two patients revealed PPR on the second EEG, and the remaining patient showed no PPR. Rapid changes in color are believed to be responsible for the photosensitive seizures because all four patients had seizures at around 18:50, when seconds of deep red and bright blue flashes, alternating at a frequency of 12 Hz, were shown. CONCLUSIONS: Regulations for technical aspects of children's programming, including the use of colors, are urgently needed in Japan to prevent a repeated incident. In addition, the IPS procedure needs to be standardized, especially for patients who are suspected to have photosensitivity. PMID- 9860072 TI - ILAE-defined epilepsy syndromes in children: correlation with quantitative MRI. AB - PURPOSE: The role of quantitative magnetic resonance imaging (MRI) in evaluation of childhood epilepsy remains poorly defined, with minimal published data. Previous work from our center questioned the specificity of hippocampal asymmetry (HA) in an outpatient group whose epilepsy was defined by using clinical and interictal data only. By using childhood volunteer controls and defining epilepsy syndromes using video-EEG monitoring, we readdressed the utility of HA in differentiating mesial temporal lobe epilepsy (MTLE) from other partial and generalized epileptic syndromes in children. METHODS: Seventy children were enrolled; entry criteria were age younger than 18 years with predominant seizure type recorded on video-EEG telemetry with volumetric MRI in all cases. Thirty healthy child volunteers had volumetric MRI. Epilepsy syndrome classification was according to ILAE. RESULTS: Control data revealed symmetric hippocampi, mean smaller/larger ratio of 0.96 (0.95-0.97, 95% CI) with no gender or right/left predominance. Overall 23% of patients had significant HA. Mean hippocampal ratio for MTLE was 0.78 (95% CI, 0.70-0.86), significantly lower than controls and from all other epilepsy syndromes. HA was highly specific (85%) to the syndrome of MTLE. Other potential epileptogenic lesions were found in 27 (39%) patients, lowest yield in frontal and mesial temporal syndromes. Dual pathology was present in 10% of patients. There was no significant association between HA and risk factors. CONCLUSIONS: In this study, we found that HA in children with a well defined epilepsy syndrome is highly sensitive and specific for MTLE. Whether this will correlate with surgical outcome, as in adults, is the subject of ongoing study. PMID- 9860073 TI - An automated injection system (with patient selection) for SPECT imaging in seizure localization. AB - PURPOSE: Ictal single-photon-emission computed tomography (SPECT) provides lateralization but has technical limitations: (a) a "truly ictal" injection must be shortly after seizure onset; (b) therefore, a seizure of brief duration may be missed; (c) more than one patient may need testing at any given time; (d) a trained health professional must stay next to each patient to inject; and (e) because the radionuclide is placed in the syringe in advance of the injection, decay of the radioactive element could result in less than optimal uptake, if the same volume of material were to be used regardless of the time after ligand preparation. METHODS: We developed an automated method of ligand injection that shortens time and increases efficiency of ictal SPECT ligand injection. By using an experimental setup, we compared manual injection times with times using an automated injection system. We determined relative costs and efficiency in work hours for the manual and automated methods. RESULTS: Injection times were 8-14 s with automated versus 19-26 s with manual injection. Readjusting volume for "ligand" decay was simple and accurate with the automated system. Injection times for clinical SPECT studies in three patients were 13, 13, and 12 s, respectively. The price of one pump equals 120 work hours of a nurse or 24 ictal injection attempts. Much of the nurse's time is "wasted" because no seizure occurs. CONCLUSIONS: The method can be more efficient of staff, shorten injection time, and facilitate obtaining "truly ictal" injections. It allows more cost-effective use of personnel. PMID- 9860074 TI - Ictal smile. AB - PURPOSE: Smiling is sometimes manifested during partial seizures. Its value for localizing the epileptogenic focus is not known. We analyzed smiling as an ictal manifestation possibly useful for seizure localization. METHODS: We reviewed patients referred to the video-EEG monitoring unit who presented a smile as part of their critical symptoms. Ictal smile was defined as an accordant expression accompanied by other characteristic epileptic symptoms and ictal EEG activity. RESULTS: Five of 86 patients experienced partial seizures with an ictal smile. We observed smiling during parietal (two patients) and temporal lobe (three patients) seizures. The right hemisphere appeared to be involved with greater frequency. CONCLUSIONS: Ictal smile is an uncommon manifestation of partial seizures involving temporal or parietal lobes, localized mainly on the right hemisphere. PMID- 9860076 TI - De novo status epilepticus as the presenting sign of neurosyphilis. AB - Although the incidence of seizures in neurosyphilis ranges from 14 to 60%, status epilepticus (SE) as a presenting complaint of neurosyphilis is definitely rare. A 44-year-old human immunodeficiency virus (HIV)-negative man with no history of epilepsy suddenly presented with acute mental confusion and was diagnosed as having a de novo complex partial nonconvulsive SE. Cerebrospinal fluid (CSF) findings, neuroimaging, and clinical course indicated that SE was the presenting symptom of an undiagnosed syphilitic meningovasculitis. The case is presented with a review of previous reports to emphasize the differential features and to underscore the importance of considering neurosyphilis among the possible causes of de novo SE. PMID- 9860075 TI - Excision of cortical dysplasia in the language area with use of a surgical navigator: a case report. AB - PURPOSE: We have developed an intraoperative optical tracking-based navigational system that allows localization in the operative space. Using three-dimensional reconstruction, this system has provided precise spatial information for intraoperative cortical mapping in patients with intractable epilepsy in whom the lesion lies close to eloquent cortex. METHODS: A 23-year-old man with intractable complex partial seizures (CPS) presented to our institution. Proton-density magnetic resonance imaging (MRI) showed a 3-cm lesion which lay 2 cm beneath the left frontal operculum. A three-dimensional model of the patient was reconstructed using MR modalities. Intraoperatively, subdural grid and strips were placed over the lesion and their electrodes were registered to the three dimensional model, which was displayed on a monitor. The navigational system was used to localize each electrode on the three-dimensional model. By the second operation, the sites of seizure activity were established and recorded on the three-dimensional model. A bipolar stimulator was also used to determine the speech area. RESULTS: The lesion, which proved to be cortical dysplasia, was removed completely and the cortical speech area was avoided. During the postoperative period, the patient had no neurological symptoms and no seizure activity. CONCLUSIONS: The localization of a lesion and its correlation with epileptogenic foci is important in optimizing treatment in patients with cortical dysplasia. Our navigational system provided accurate localization of the lesion and correlation with the epileptogenic focus and related eloquent cortex. We believe that the safe removal of the lesion was facilitated by this system. PMID- 9860077 TI - Abdominal epilepsy in an adolescent with bilateral perisylvian polymicrogyria. AB - PURPOSE: We report a patient with recurrent episodes of severe periumbilical pain accompanied by headache, pallor, dizziness, and visual hallucinations who was subsequently diagnosed as having abdominal epilepsy and a bilateral sylvian cortical malformation. METHODS AND RESULTS: During an EEG examination, the patient had a simple partial seizure, manifested as intense abdominal pain. The ictal EEG showed a focal electrographic seizure arising from the left frontotemporal region. Computed tomography scan was normal; however, magnetic resonance imaging (MRI) revealed asymmetric lesions involving both sylvian fissures compatible with polymicrogyria. Onset of treatment with sodium valproate was followed by a significant reduction of seizures. CONCLUSIONS: Abdominal epilepsy should be suspected in adolescents or adults with paroxysmal episodes of abdominal pain associated with migraine-like symptoms. EEG and MRI investigations are indicated. A developmental brain disorder should be considered as a possible etiology. PMID- 9860078 TI - Guidelines for neuroimaging evaluation of patients with uncontrolled epilepsy considered for surgery. Commission on Neuroimaging of the International League Against Epilepsy. PMID- 9860079 TI - Clinical and MRI evidence that occipital lobe seizures can be the major manifestation of the reversible posterior leukoencephalopathy syndrome (RPLS) PMID- 9860080 TI - Hypothesis that tiagabine-induced NCSE is associated with GABAergic hyperfunction, with GABA(B) receptors playing a critical role, is supported by a case of generalized NCSE induced by baclofen. PMID- 9860081 TI - Comprehensive asthma management: guidelines for clinicians. AB - Asthma is a chronic inflammatory disease characterized by reversible airway obstruction and nonspecific airway hyperreactivity. Asthma is managed in steps according to disease symptoms and severity. Treatment goals are to decrease symptoms, improve pulmonary function, and reduce overall morbidity and the associated cost of medical care. Antiasthma drugs are a key component of asthma management that are classified as either long-term-control medications that control symptoms and prevent disease exacerbations, or quick-relief medications that rapidly relieve airway obstruction and acute asthma symptoms. Several new leukotriene (LT) modulators have been developed that promise to improve asthma control, including LT receptor antagonists montelukast and zafirlukast and the 5 lipoxygenase inhibitor zileuton. Each decreases symptoms and the use of rescue medication, and improves pulmonary function in patients with mild intermittent to moderate persistent asthma. PMID- 9860082 TI - Chronic childhood illness and maternal mental health--why should we care? AB - This study evaluated which factors most influenced the impact of childhood asthma on the child's family. Seventy children/families seen at a tertiary-care hospital for asthma were evaluated. Stepwise multiple regression examined the effects of illness severity; family socioeconomic status (SES); family structure; social support; child's emotional characteristics; parent's health; family functioning; and maternal psychological distress on the Family Impact of Illness Scale. Analysis indicated that only the parent's Psychiatric Symptom Index significantly predicted impact scores. The most important predictors of how much impact a child's asthma has on the family are parental emotional distress and amount of social support. PMID- 9860083 TI - Predictors of treatment adherence among asthma patients in the emergency department. AB - This study examined predictors of treatment adherence among 120 adult patients with asthma at two emergency departments (EDs). Structured medical chart reviews were performed for characteristics hypothesized to be associated with treatment adherence difficulties. Sixty percent of subjects had evidence of nonadherence with asthma treatment. Several variables were associated with nonadherence including younger age, more utilization of ED services (more ED visits, using the ED for medication refills), certain treatment characteristics (receiving more medications in the ED, not being prescribed prednisone at discharge), and not keeping post-discharge follow-up appointments. Further research should be directed at enhancing self-management skills and decreasing inappropriate ED use among nonadherent patients. PMID- 9860084 TI - Adherence-related behavior in adolescents with asthma: results from focus group interviews. AB - Focus group interviews were conducted with 14 adolescents with asthma to explore self-management behavior, in particular with regard to adherence behavior. In addition, the adolescents discussed their feelings about having asthma, gave insight into how they evaluate the provided health care, and made recommendations for healthcare providers and for the development of patient education materials. The majority of participants did not take their prophylactic asthma medication regularly, and were rather late in starting to use their bronchodilator. They were sometimes fed up with having asthma. Moreover, the majority of participants were not always frank in telling their pediatrician how they managed their asthma. Finally, they found it essential that information about asthma should be given personally and not by means of leaflets, and recommended that healthcare providers should use audio-visual aids to illustrate what they are explaining. The results of the focus group interviews have been used for the development of an intervention program which aims at enhancing adherence in adolescents with asthma. PMID- 9860085 TI - High-dose inhaled budesonide may substitute for oral therapy after an acute asthma attack. AB - Patients attending the emergency room with acute asthma, participating in a study comparing salbutamol (albuterol in the United States) via a dry powder inhaler (Turbuhaler) with pressurized metered-dose inhaler (pMDI), were included in this 1-week follow-up study with the aim of assessing whether inhaled budesonide via Turbuhaler may be an alternative to prednisolone tablets after an acute asthma attack. Eighty-one patients with a mean age of 38 years and forced expiratory volume in 1 sec (FEV1) of 64% predicted normal value after treatment with salbutamol were randomized in this double-blind, double-dummy, parallel-group study. The doses given were budesonide 1600 microg b.i.d. or prednisolone in daily doses from 40 mg (day 1) decreased to 5 mg (day 7). FEV1 was recorded before and after the 7-day treatments and peak expiratory flow (PEF) morning and evening, clinical symptoms (visual analogue scale 0-100), and doses of rescue medication (terbutaline Turbuhaler 0.25 mg/dose) were recorded daily. The mean increase in FEV1 from baseline to day 7 was 17.3% in the budesonide Turbuhaler group and 17.6% in the prednisolone group. Mean values of morning PEF increased from day 1 to day 7 by 67 L/min in the budesonide Turbuhaler group and by 57 L/min in the prednisolone group (not significant). There were no statistically significant differences between the groups in clinical symptoms and in the number of doses of rescue medication. Because of disease deterioration, five patients in the Turbuhaler group and three in the prednisolone group needed additional symptomatic as well as corticosteroid treatment. Inhaled budesonide in high doses may be a substitute for oral therapy as follow-up treatment after an acute asthma attack. PMID- 9860087 TI - Complementary therapies for asthma: what patients use. AB - Asthma patients are prime candidates to employ complementary medicine (CM) for their conditions, yet little hard data exist on the prevalence of CM use by these patients. This survey was aimed at generating some information on this and related issues. Members of the UK National Asthma Campaign (17,000) were invited to complete a questionnaire and 4741 responded. Only 41% of the respondents had never tried CM. Breathing techniques, homoeopathy, and herbalism were the three most prevalent treatments. The majority of respondents perceived these therapies as moderately useful with only little difference between various treatments. In general, the money spent on CM was modest. It is concluded that CM is frequently used for asthma. Therefore, this area of healthcare would seem to warrant more rigorous investigation. PMID- 9860086 TI - Features that distinguish those who die from asthma from community controls with asthma. AB - To evaluate risk factors for asthma mortality, an unmatched case-control study was undertaken in the Canadian prairie provinces of Alberta, Saskatchewan, and Manitoba. Those between the ages of 5 and 50 (inclusive) who died from an acute exacerbation of asthma were compared to a control group of people with asthma from the same geographical areas who were contacted using random-digit dialing. Because no deaths occurred among residents less than 15 years old, this analysis was limited to cases and controls between 15 and 50 years old. Of the 38 deaths that occurred between November 1992 and October 1995, data were obtained from next of kin for 35 (92.1%). Of the 210 potential controls that were identified, 142 returned completed questionnaires (67.6%). Cases were more likely than controls to have asthma reported to be severe, to have experienced nocturnal symptoms, to have had cardiopulmonary resuscitation (CPR)/intubation, and to have had more healthcare utilization in the previous year. Medication use was also more common among cases compared to controls. Specific asthma triggers were reported more often for cases than controls; weather changes, excitement, depression, and stress showed the greatest case control differences. Although a number of very strong risk factors for death from asthma were identified, death from asthma is so rare in this age group that it is not possible to label an individual as "likely" to die from asthma. Nonetheless, patients, caregivers, and health professionals should be aware of indicators that would suggest greater risk. PMID- 9860088 TI - Comparative analysis of calcium-binding protein-immunoreactive neuronal populations in the auditory and visual systems of the bottlenose dolphin (Tursiops truncatus) and the macaque monkey (Macaca fascicularis). AB - This study compares the distribution of three calcium-binding protein immunoreactive (CaBP-immunoreactive) neuronal populations (calretinin-, calbindin and parvalbumin-immunoreactive) in the visual and auditory systems in two mammalian species which are fundamentally different in their evolutionary traits and ecology, the aquatic toothed whale Tursiops truncatus (bottlenose dolphin) and the terrestrial Old World primate, Macaca fascicularis (long-tailed macaque). Immunocytochemical analyses, combined with computerized morphometry revealed that in the visual and auditory systems of the bottlenose dolphin, calretinin and calbindin are the prevalent calcium-binding proteins, whereas parvalbumin is present in very few neurons. The prevalence of calretinin and calbindin immunoreactive neurons is especially obvious in the auditory system of this species. In both auditory and visual systems of the macaque monkey, the parvalbumin-immunoreactive neurons are present in comparable or higher densities than the calretinin and calbindin-immunoreactive neurons. In some structures of the visual and auditory systems of the macaque monkey, the calretinin- and calbindin-immunoreactive neurons are nearly absent. The prevalence of parvalbumin immunoreactive over calretinin- and calbindin-immunoreactive neurons is particularly prominent in the visual system of primates. Thus, the dominant sensory systems in both aquatic and terrestrial mammals are enriched in specific phenotypes of calcium-binding protein-immunoreactive neurons. PMID- 9860089 TI - Calcium binding protein calcyphosine in dog central astrocytes and ependymal cells and in peripheral neurons. AB - Calcyphosine is a calcium binding protein discovered in the dog thyroid in 1979. Calcyphosine mRNA and immunoreactivity were detected using Western and Northern blotting in the cerebral cortex, cerebral white matter and cerebellum. Using immunohistochemistry and in situ hybridization, both are present in ependymal cells, choroid plexus cells and several types of astrocytes of the subependymal cerebral layer, the cerebellar Bergmann layer, the retinal ganglion cell layer, the optic nerve and the posterior pituitary. Both are also present in neurons of nasal olfactory mucosa, enteric Auerbach and Meissner plexuses, orthosympathic and spinal cord ganglia as well as in endocrine cells of neural crest origin in the adrenal medulla. Calcyphosine immunoreactive astrocytes were also present mainly in hemispheric cerebral gray and white matter, hemispheric subcortical structures, brain stem and spinal cord. These results show that calcyphosine is a characteristic calcium binding protein of astrocytes and ependymal cells in the central nervous system and of neurons in the peripheral nervous system. This is of interest in view of the importance of calcium regulation in these cells, and since calcyphosine a calcium binding protein phosphorylated by cAMP dependent process, may be an intermediate between cAMP and inositol phosphate cascades. PMID- 9860090 TI - Presence of galanin in dopaminergic neurons of the sheep infundibular nucleus: a double staining immunohistochemical study. AB - The distribution of tyrosine hydroxylase (TH) and of galanin immunoreactive (IR) neurons were examined in the sheep infundibular nucleus. Antisera raised against TH and galanin were used on adjacent sections and for double immunohistochemical staining of the same sections. There was considerable overlap in the distribution of TH and galanin-IR neurons in the medial part of the nucleus. Most of the galanin-IR neurons were also TH-IR, but less than 50% of the TH-IR neurons also expressed galanin immunoreactivity. Neurons immunoreactive to TH alone were observed close to the third ventricle and in the rostral part of the infundibular nucleus. In the median eminence, TH and galanin-IR fibres overlapped mainly in the lateral and dorsal parts of the external layer, but the colocalisation of both antigens could not be assessed on the available material. Thus, in sheep, the population of catecholaminergic neurons of the infundibular nucleus may be subdivided into different subpopulations according to their peptide content, but does not appear segregated as in rat and human. PMID- 9860091 TI - Antimicrobial function of human saliva--how important is it for oral health? AB - Human saliva contains a number of physical physicochemical, and chemical agents that protect oral tissues against noxious compounds, in particular those produced by various microorganisms. Among such protective factors, the flushing effect of saliva flow is the most important one, not only because it so effectively removes exogenous and endogenous microorganisms and their products into the gut but also because a steady supply of saliva guarantees continuous presence of both non immune and immune factors in the mouth. A great number of studies with controversial results have been published regarding various individual agents and their possible association to oral health, particularly to dental caries. It appears that no single chemical agent is far more important than the others. For example, patients with selective IgA deficiency have normal levels of non-immune defense factors and often display a compensatory increase in the other immunoglobulin isotypes. The concerted action of all agents in whole saliva, both saliva- and serum-derived, provides a multifunctional protective network that is collapsed only if salivary flow rate is substantially reduced. In this mixture of defense factors, many show additive or even synergistic interactions against oral pathogens. Increased knowledge of the molecular functions of various agents has made it possible to prepare oral hygiene product that include host-derived antimicrobial agents instead of synthetic agents. Although the clinical efficacy of such products is still unsatisfactory and poorly described, new technologies, for example in the production of specific antibodies against oral pathogens, may considerably improve the antimicrobial power of these products. PMID- 9860092 TI - Shear stresses in the adhesive layer under porcelain veneers. A finite element method study. AB - A finite element study was made investigating the shear stresses in the composite cement layer and the enamel bond under loaded porcelain veneers. The maximum shear stresses were calculated under various conditions concerning loading angle, laminating extension, and cervical design. The result of the study was that maximum shear stresses did not exceed the stress level for debonding, but that great differences in maximum shear stress appeared with varying loss of bond and different loading angle. The fully laminated facing showed stress levels in the composite cement only 1/5 of those in the facing with a lack of adhesion in the periphery, and 1/15 in the enamel bond. The maximum stresses increased about 4 times when the load angle was 30 degrees as compared with 0 degrees, and increased about 1.5 times from 30 degrees to 60 degrees. The importance of getting a full lamination of the veneer and avoiding unfavorable loading conditions is emphasized in this study. PMID- 9860093 TI - A 5-year clinical evaluation of ceramic inlays (Cerec) cemented with a dual-cured or chemically cured resin composite luting agent. AB - Sixty-six class-II CAD/CAM-manufactured ceramic inlays (Cerec) were placed in 27 patients. Each patient received at least one inlay luted with a dual-cured resin composite and one inlay luted with a chemically cured resin composite. The inlays were examined 5 years after luting using the California Dental Association (CDA) criteria. Eighty-nine percent of the 66 inlays were rated 'satisfactory'. During the follow-up period replacement was required for 3 inlays because of inlay fractures (4.5%) and 1 inlay because of fracture of the tooth substance (1.5%). All those inlays were luted with the dual-cured resin composite luting agent. Of the remaining 62 inlays the CDA rating 'excellent' was given to 84% for color, 97% for surface, and 81% for anatomic form. 'Excellent' margin integrity was seen in 52% of the dual-cured resin composite luted inlays and in 61% of the chemically cured resin composite luted inlays. No statistically significant (P> 0.05) difference was observed between the two luting agents. PMID- 9860094 TI - Inductive analysis methods applied on questionnaires. AB - The am of this study was to evaluate subjective aspects from questionnaires dealing with dental trauma by applying different computerized inductive techniques within the field of artificial intelligence to questionnaires consisting of descriptive variables and of questions reflecting functional, personal, and social effects of patients' oral situation following dental trauma. As the methodology used is new to many readers in odontologic sciences, a detailed description of both the processes and the terminology is given. Utilizing a neural network as a first step in an analysis of data showed if relations existed in the training set, but the network could not make the relations explicit, so other methods, inductive methods, had to be applied. Inductive methods have the potential constructing rules from a set of examples. The rules combined with domain knowledge can reveal relations between the variables. It can be concluded that the usage of methods based on artificial intelligence can greatly improve explanatory value and make knowledge in databases explicit. PMID- 9860095 TI - Effects of continuous glucocorticoid infusion on the progression of dentinal caries in growing rats. AB - This study was undertaken to test the effects of a low dose of continuous glucocorticoid infusion on the rate of progression of dentinal caries in molars of young rats. Forty-seven rats were inoculated in the mouth with Streptococcus sobrinus and fed ad libitum a cariogenic diet and 10% sweetened water. After 10 days of caries initiation ten animals were killed to serve as a reference group. In the rest of the animals the cortisone or placebo pellet was implanted subcutaneously in the back of the neck. The daily release of cortisone was 0.42 mg per rat. Sweetened water was changed to pure water, and the diet was the same cariogenic diet. After 6 weeks of medication the areas of dentinal caries were quantified planimetrically. Schiff's staining was used to classify caries. Although cortisone medication slightly increased the number of carious lesions, statistical significance was not reached. However, compared with the placebo group, the rats receiving cortisone medication showed significantly increased dentinal caries progression and severity of lesions. This study suggests that glucocorticoids with a cariogenic diet reduce the intrinsic modulation or response of the odontoblasts to caries attack. PMID- 9860096 TI - The gingival plasma cell infiltrate in renal transplant patients on an immunosuppressive regimen. AB - Treatment with immunosuppressive agents inhibits gingival inflammation and progression of periodontitis in humans. We examined the numbers and the isotype distribution of immunoglobulin-producing plasma cells by immunohistochemistry in gingival specimens taken from renal transplant transplant recipients receiving immunosuppressive agents (IS), and from otherwise comparable systemically healthy patients. The immunosuppressed patient group had significantly (P< 0.05) fewer IgG-, IgA-, IgG1-, IgG2-, and IgG4-producing plasma cells in the connective tissue adjacent to the pocket epithelium. The reduced numbers of such patents with quiescent periodontal disease support the contention that high counts of plasma cells are indicative of more severe disease. PMID- 9860097 TI - Quality evaluation of oral health record-keeping for Finnish young adults. AB - The objective of this study was to assess the quality of oral health record keeping in public oral health care in relation to dentists' characteristics. A random computerized selection of 239 subjects, born in 1966-71 and clinically examined during 1994 in an administrative unit of the public oral health service in southern Finland, included 4-5 cases per dentist, the number of dentists being 50. Data concerning actual clinical examinations and treatment courses carried out in public dental clinics came from original oral health records. Criteria for assessment of oral health record entries were based on Finnish health legislation and detailed instructions of health authorities. The results showed that each patient's identity was available in 90% of documents. Recordings concerning continuity of comprehensive care were infrequent; a questionnaire concerning each patient's up-to-date health history was in only 26% of the oral health records. Notes concerning each patient's bite and function of the temporomandibular joint were in 37% of the records, notes about oral soft tissues were in 11%, and the check-up interval was recorded in 21%. Recording of indices on periodontal and dental status varied greatly; the community periodontal index of treatment need was found in 93% and the index of incipient lesions in 16% of the records. Female dentists and dentists younger than 37 years tended to record more information. Dentists should be encouraged to better utilize the options offered by oral health records for individual treatment schemes. PMID- 9860098 TI - Comparison of dentin apposition and dentinal caries progression in the mandibular and maxillary molars of the rat. AB - To show that the rate and the rate of reduction of dentin apposition are about the same in mandibular and maxillary molars, 23 Sprague-Dawley rat pups were randomized into 2 groups on the day of birth. During lactation half of their dams received a standard rodent diet; the other half, a diet containing 41% sucrose. At the age of 3 weeks the pups were weaned, weighed, given an intraperitoneal injection of oxytetracycline hydrochloride, and inoculated with oral Streptococcus sobrinus. During the experiment the pups received the same diet as their dams during lactation. After 5 weeks the pups were decapitated, their jaws sectioned sagittally, the first and second molars photographed, and the areas of dentin apposition and dentinal caries measured planimetrically. The area of dentin formation was about the same in maxilla and mandible in the first molars, but slightly smaller in the mandibular second molars of the control group. The sucrose diet reduced dentin apposition significantly in both jaws, although the areas were significantly smaller in the mandibles than in the maxillae. Caries did not affect the rate of dentin apposition. The areas of caries lesions were smaller in the maxillary molars of both diet groups. The results show that the hypothesis of equal rate of dentin apposition in mandible and maxilla was not valid because the reduction, caused by sucrose, was more prominent in mandibular molars for unknown reasons. The reduction of dentin apposition was reflected as acceleration of caries progression among the diet groups and the jaws. It was concluded that the response of the pulpodentinal complex to sucrose and dentinal caries during the primary dentinogenesis cannot be seen as a formation of reactionary or reparative dentin, as with adult rats. PMID- 9860099 TI - Perception of treatment need among orthodontic patients compared with professionals. AB - The aim was to evaluate estimated need for orthodontic treatment, as judged from intraoral photographs, among orthodontic patients and professionals. Twenty consecutive prospective orthodontic patients, 20 consecutive orthodontically treated patients, 10 randomized general dentists, and 10 orthodontists participated. Seventy pairs of anonymous intraoral photographs of dentitions with varying degrees of objective treatment need were randomly arranged in a notebook. The general dentists and orthodontists rated orthodontic treatment need on a visual analog scale in a similar way among themselves and were more reserved than both patient categories, who also scored similarly among themselves. Professional raters also had similar inter- and intra-rater reliability among themselves, and it was higher than in either of the patient categories. Treatment providers appear to be more restrictive, consistent, and reliable in their judgement of orthodontic treatment need from intraoral photographs than the target group, patients positive toward orthodontic treatment. PMID- 9860101 TI - The association between dental arch dimensions and occurrence of Finnish dental consonant misarticulations in cleft lip/palate children. AB - The aim of this study was to examine whether maxillary and mandibular dental arch width, length, and palatal height dimensions are associated with the occurrence of misarticulations (phonetic or phonologic errors) in the dental consonants /r/, /s/, and /1/ in different cleft types and sexes. The subjects were 263 (109 girls, 154 boys) 6-year-old Finnish-speaking non-syndromic children with isolated cleft palate (CP, n=79), deft lip/alveolus (CL(A), n=77), unilateral (UCLP, n=80), and bilateral (BCLP, n=27) cleft lip and palate. Dental plaster casts were measured by two authors using the technique of Moorrees, and auditive speech was analyzed with high reliability by two speech pathologists. The results showed that the occurrence of misarticulations increased and dental arch dimensions decreased with the severity of the cleft. Narrower and shorter maxillary arches as well as shallower palates were related to problems with the studied dental consonants. Mandibular arch dimensions were not related to the misarticulations. However, statistical analysis did not reveal significant differences in dental arch dimensions between subjects with and without misarticulations when they were compared separately for different cleft types. The etiology of clefting per se- isolated deft palate versus cleft lip with or without deft palate--did not seem to explain the associations between dental arch dimensions and the studied misarticulations. PMID- 9860100 TI - Odontologic survey of referred patients with symptoms allegedly caused by electricity or visual display units. AB - Twenty-eight consecutive patients with symptoms allegedly caused by electricity or visual display units were odontologically investigated according to a specially designed registration form including an anamnestic interview and a clinical protocol. The most common oral and general symptoms reported were burning mouth, craniomandibular dysfunction symptoms, skin complaints, and fatigue. Oral symptoms such as craniomandibular dysfunction and general symptoms such as eye complaints and dizziness scored highest on a visual analog scale regarding mean symptom intensity. The patients reported various numbers of medical diagnoses, such as allergic rhinitis or asthma and hypothyroidism. Various dental diseases were found; the most common were temporomandibular joint and masticatory muscle dysfunctions, lesions in the oral mucosa, and periodontal diseases. Urinary-Hg (U-Hg) analysis showed a mean U-Hg concentration of 8.5 nmol Hg/L urine, and none of the patients exceeded the limit of 50 nmol Hg/L urine. The U-Hg concentration was positively correlated with the number of amalgam fillings (P< 0.01) and craniomandibular disorders (P < 0.05). No or low secretion of the minor mucous glands was found in 43% of the patients. One patient showed hypersensitivity to gold and cobalt. The present study showed that various odontologic factors might be involved in some of these patients' suffering. Thus, it is important that professionals from other disciplines collaborate with dentistry if these patients are to be properly investigated. PMID- 9860102 TI - The effect of chronic administration of antidepressants on the circadian pattern of corticosterone in the rat. AB - Although antidepressant administration has been reported to alter the pituitary adrenal (PA) axis, the results are puzzling. In the present work, two possible factors contributing to these contradictory results were studied in adult male Sprague-Dawley rats: (i) the type of antidepressant and (ii) the time of day at which samples were taken. Samples were taken under nonstressful conditions. In expt 1, the acute effects of two doses (10 and 20 mg/kg) of the tricyclics clomipramine (CMI), desipramine (DMI) and imipramine (IMI), and the non-specific monoamine oxidase inhibitor (MAOI) phenelzine were studied. Only phenelzine increased plasma corticosterone with the low dose, whereas phenelzine and DMI increased plasma corticosterone with the high doses when measured 30 min after drug administration. In a second experiment, it was observed that after 12 daily doses of the drugs (20 mg/kg), all drugs increased plasma corticosterone at 30 min after the last drug administration. When the circadian pattern of corticosterone was studied in the same experiment, starting on the day after the last drug administration, a significant interaction of drug by time of day was found. Drugs caused changes in the normal levels of plasma corticosterone at certain times and DMI, IMI and phenelzine reduced the number of rats showing the normal corticosterone peak at 1900 hours. No significant effect of drugs on corticosteroid-binding globulin (CBG) was found. In a third experiment, phenelzine and IMI were administered as before, but samples were taken at several times both on the day of the last drug administration and on the following day. The two drugs altered the normal circadian pattern of corticosterone in a somewhat different way, but both caused a reduction of the corticosterone peak at lights off on the day after the last drug administration. The normal relative thymus weight observed in all groups (exp. 2) suggests that the overall biological activity of corticosterone was probably not affected by antidepressants. The present results indicate that most antidepressants are able acutely to activate the PA axis after repeated administration in a similar way or even more-strongly than after the first administration, and that some of these drugs alter normal circadian pattern of corticosterone. No evidence for decreased resting PA activity was found in antidepressant-treated rats. PMID- 9860103 TI - Transdermal nicotine effects on attention. AB - Nicotine has been shown to improve attentiveness in smokers and attenuate attentional deficits in Alzheimer's disease patients, schizophrenics and adults with attention-deficit/hyperactivity disorder (ADHD). The current study was conducted to determine whether nicotine administered via transdermal patches would improve attentiveness in non-smoking adults without attentional deficits. The subjects underwent the nicotine and placebo exposure in a counterbalanced double-blind manner. Measures of treatment effect included the Profile of Mood States (POMS), Conners' computerized Continuous Performance Test (CPT) of attentiveness and a computerized interval-timing task. The subjects were administered a 7 mg/day nicotine transdermal patch for 4.5 h during a morning session. Nicotine significantly increased self-perceived vigor as measured by the POMS test. On the CPT, nicotine significantly decreased the number of errors of omission without causing increases in either errors of commission or correct hit reaction time. Nicotine also significantly decreased the variance of hit reaction time and the composite measure of attentiveness. This study shows that, in addition to reducing attentional impairment, nicotine administered via transdermal patches can improve attentiveness in normal adult non-smokers. PMID- 9860104 TI - The influence of nicotine pretreatment on mesoaccumbens dopamine overflow and locomotor responses to D-amphetamine. AB - Pretreatment with psychostimulant drugs causes sensitisation of their effects on locomotor activity and dopamine (DA) overflow in the nucleus accumbens (NAcc) and there is evidence for similarities in the mechanisms involved. This study used in vivo microdialysis in conscious freely moving rats to investigate the extent to which pretreatment with nicotine causes sensitisation to D-amphetamine. Pretreatment with nicotine (0.4 mg/kg s.c. daily for 5 days) caused sensitisation of the locomotor responses to D-amphetamine (0.1-0.5 mg/kg s.c.) but not cocaine (15 mg/kg i.p.). Nicotine pretreatment did not influence the increase in DA overflow into dialysis probes, located in the core of the NAcc, evoked by systemic injections of D-amphetamine or cocaine (15 mg/kg i.p.) but decreased the overflow evoked by the administration of D-amphetamine (1 x 10(-6) M) through the dialysis probe. The results provide further evidence for a dissociation between the expression of sensitised locomotor responses to psychostimulant drugs and sensitisation of their stimulatory effects on DA overflow in the core of the NAcc. The results suggest that the sensitisation of the effects of nicotine on DA overflow in this subdivision of the NAcc may be pharmacologically specific to nicotinic drugs. PMID- 9860105 TI - Role of the retrorubral nucleus in striatally elicited orofacial dyskinesia in cats: effects of muscimol and bicuculline. AB - Orofacial dyskinesia (OFD) is a disorder characterized by involuntary movements of the oral and facial muscles. OFD attacks can be elicited acutely in cats by local injections of dopaminergic agents into the anterodorsal part (r-CRM) of the caudate nucleus. Because the dopaminergic A8 cell group, being embedded in the retrorubral nucleus (RRN), gives rise to fibres which terminate in the r-CRM, two questions arose: (1) whether the A8 cell group forms part of the circuitry that directs and/or modulates OFD, and (2) whether GABA-ergic compounds in the RRN play a role in OFD, and if so, whether a pharmacological GABA-ergic intervention of the activity in the RRN modulates or mediates OFD. For this purpose, the activity of the RRN was manipulated with local injections of the GABA(A) agonist muscimol and antagonist bicuculline. These local injections into the RRN were subsequently combined with manipulations of dopamine transmission in the r-CRM with local injections of the selective DAi receptor agonist (3,4 dihydroxyphenylimino)-2-imidazoline. The present study shows that local injections of GABA-ergic compounds into the RRN do not elicit OFD attacks in cats, but can modulate oral behaviour elicited from the r-CRM. The latter effect is dose dependent and GABA-ergic specific. PMID- 9860106 TI - Impairment of memory and plasma flunitrazepam levels. AB - Flunitrazepam was administered to volunteers in three different oral doses. The effects on psychomotor sedation, attention, working memory and explicit memory were then assessed at various intervals after dosing and compared with levels of the drug in the plasma. Three groups of 12 healthy males with similar levels of education were given placebo or flunitrazepam (1, 2 or 4 mg) in a double-blind, random-sequence study. Volunteers completed a battery of tests at night, 3.5 h after taking the drug and in the morning, 10 h afterward. Blood samples were collected for drug analysis before and after the nocturnal tests and before morning tests. At night, only the highest dose of flunitrazepam (4 mg) induced significant changes in psychomotor sedation, attention, working memory, and prose immediate recall. Doses of 2 and 4 mg flunitrazepam significantly reduced the mean scores of explicit memory (morning tests). Z-scores, calculated from differences between flunitrazepam and placebo, revealed that 2 mg flunitrazepam impaired memory but not alertness or attention. Linear regression analysis of the relationship between plasma levels of flunitrazepam and its effects (Z-scores) indicated that there was a significant positive correlation between peak levels of flunitrazepam at night and impairment of night attention and explicit memory, i.e. delayed recall of prose (r = 0.59, P < 0.01) and trigrams (r = 0.55, P < 0.01). However, memory and attention Z-scores as a function of plasma levels fitted with nonlinear regression analysis to the Emax model had higher correlation coefficients. To produce an effect equal to 50% of the maximum effect for memory impairment, concentrations (EC50) were 6.1 and 6.4 ng/ml for prose and trigrams delayed recall; but for attention they were much higher, at 13.2 ng/ml. The overall results indicate that higher concentrations were needed to impair attention than were required to impair memory. PMID- 9860107 TI - Clinical pharmacology of moclobemide during chronic administration of high doses to healthy subjects. AB - The objectives of this study were to assess the tolerability, safety, pharmacodynamics and pharmacokinetics of high-dose moclobemide in healthy subjects. Two sequential groups of six male and six female subjects (eight on active treatment, four on placebo) received for 8 days moclobemide 450 mg b.i.d. and 600 mg b.i.d., respectively. Intravenous tyramine pressor tests were conducted at baseline, at the beginning of treatment and at steady state. Oral tyramine pressor tests with 50, 100 and 150 mg tyramine were conducted under steady-state conditions. Pharmacokinetic parameters of moclobemide and two of its metabolites in plasma and urine were determined after the first and last dose of moclobemide. The incidence and intensity of adverse events was dose-dependent. The most frequently reported adverse events were insomnia, headache, dizziness and dry mouth. The i.v. tyramine pressor sensitivity during both moclobemide dosing regimens was enhanced 3 to 4-fold. Intake of tyramine 50 mg did not result in systolic blood pressure increases greater than 30 mmHg. With regard to blood pressure increases, tyramine 100 mg is still compatible with moclobemide 450 mg b.i.d. but not with 600 mg b.i.d. The clearance of moclobemide decreased by about 60% on multiple dosing, but no differences were found between both dosing regimens. The urinary excretion of the N-oxide metabolite doubled during multiple dosing. In conclusion, the maximum tolerated dose of moclobemide in healthy subjects is 600 mg b.i.d. provided the tyramine content in a meal is not higher than 50 mg. PMID- 9860108 TI - Ziprasidone 40 and 120 mg/day in the acute exacerbation of schizophrenia and schizoaffective disorder: a 4-week placebo-controlled trial. AB - A double-blind, placebo-controlled, multicenter study, was performed to evaluate the efficacy and safety of ziprasidone in 139 patients with an acute exacerbation of schizophrenia or schizoaffective disorder. Patients were randomized to receive ziprasidone 40 mg/day, 120 mg/day or placebo for 28 days. Ziprasidone 120 mg/day was significantly more effective than placebo in improving the BPRS total, CGI-S. BPRS depression cluster and BPRS anergia cluster scores (all P < 0.05). Similarly, the percentages of patients classified as responders on the BPRS (> or = 30% reduction) and the CGI improvement (score < or = 2) were significantly greater with ziprasidone 120 mg/day compared with placebo (P < 0.05). The number of patients who experienced an adverse event was similar in all three treatment groups, and discontinuation due to adverse events was rare (five of 91 ziprasidone-treated patients). The most frequently reported adverse events, that were more common in either ziprasidone group than in the placebo group, were dyspepsia, constipation, nausea and abdominal pain. There was a notably low incidence extrapyramidal side-effects (including akathisia) and postural hypotension and no pattern of laboratory abnormalities or apparent weight gain. Ziprasidone-treated patients were not clinically different from placebo-treated patients on the Simpson-Angus Rating scale, Barnes Akathisia scale and AIMS assessments. These results indicate that ziprasidone 120 mg/day is effective in the treatment of the positive, negative and affective symptoms of schizophrenia and schizoaffective disorder with a very low side-effect burden. PMID- 9860109 TI - Naltrexone administration affects ad libitum smoking behavior. AB - Endogenous opioid peptides have been implicated in the reinforcement of smoking and opioid antagonists have been examined to determine their role in smoking behavior. To date, the relationship between smoking behavior and chronic opiate antagonist administration during ad libitum smoking has not been investigated. The purpose of this study was to examine the relationships between naltrexone, an opiate antagonist administered orally, and smoking behavior and mood states during ad libitum smoking. A repeated measures experimental design was used. Normal adult male and female volunteers, admitted to the Clinical Research Center, were randomly assigned to naltrexone-treated (n = 22) or placebo-control (n = 21) groups in a double-blind manner. Day 1 was considered acclimation to the unit and day 2 was baseline, or pre-drug administration. On days 3, 4, and 5, subjects received 50 mg naltrexone or a placebo at 0700 and 1600 hours. Plasma nicotine and expired air carbon monoxide levels were measured daily at 1900 hours. Number of cigarettes smoked, mood states, withdrawal symptomatology and self-reported satisfaction with smoking were also quantified daily. Results indicated that plasma nicotine levels (P = 0.005), number of cigarettes smoked daily (P = 0.003) and self-reported satisfaction with smoking (P = 0.043) were significantly lower among those treated with naltrexone, compared to the placebo control group. Expired air carbon monoxide levels did not differ between the two groups. In addition, mood states and withdrawal symptoms did not differ between groups. These findings suggest that endogenous opioid peptides influence specific smoking behavior variables. PMID- 9860110 TI - Subjective, psychomotor, and analgesic effects of oral codeine and morphine in healthy volunteers. AB - The subjective, psychomotor, and physiological effects of analgesic doses of oral codeine and morphine were examined in 12 healthy volunteers. Subjects ingested placebo, morphine 20 or 40 mg, or codeine 60 or 120 mg in a randomized, double blind, crossover design. The smaller and larger doses of each drug were putatively equianalgesic, and the cold-pressor test was included to test this assumption. Codeine and morphine increased ratings of "feel drug effect" but had little effect on other subjective measures, including the Addiction Research Center Inventory, visual analog scales, and adjective checklists. The few subjective effects that were observed were modest and were dose-related for morphine but not for codeine. The drugs did not affect performance on Maddox Wing, digit-symbol substitution, coordination, auditory reaction, reasoning, and memory tests. Dose-related decreases in pupil size (miosis) were observed following codeine and morphine. Ratings of pain intensity decreased in a dose related manner for morphine but not for codeine. Plasma codeine and morphine levels varied as an orderly function of dose. These results suggest that oral codeine and morphine are appropriate drugs for outpatient pain relief because they are effective analgesics at doses that have only modest effects on mood, produce few side effects, and do not impair performance. The results also suggest a possible ceiling effect of codeine on analgesia and subjective effects. PMID- 9860111 TI - Both nicotine and mecamylamine block dizocilpine-induced explosive jumping behavior in mice: psychiatric implications. AB - Dizocilpine (MK-801) administration to an outbred strain of NIH Swiss mice elicits discrete episodes of explosive jumping behavior designated as "popping." This behavior may serve as a useful preclinical paradigm for the screening of potentially novel antipsychotic medications. Both nicotine and mecamylamine, a nicotinic antagonist, dose-dependently blocked dizocilpine-induced popping. The data suggest that nicotine may be of therapeutic benefit in the treatment of schizophrenia and that some of its effects may be mediated by non-nicotinic receptors. PMID- 9860112 TI - Dextrorotatory opioids induce stereotyped behavior in Sprague-Dawley and Dark Agouti rats. AB - Dextromethorphan and dextrorphan elicited a stereotyped behavioral syndrome in rats indistinguishable from that produced by PCP and other non-competitive NMDA antagonists. The rank order of potency for the induction of stereotyped behavior in male Sprague-Dawley rats was: MK-801>PCP>(+/-)cyclazocine>dextrorphan>(+/ )ketamine>dextromethorphan. These behavioral potencies were significantly correlated (0.91; P<0.05) with their respective affinities for high affinity [3H]dextrorphan-labelled NMDA receptors in rat forebrain membranes. To address the propensity of dextromethorphan to induce stereotyped behavior, dextrorotatory opioid induced stereotypies were investigated in female Dark Agouti and female Sprague-Dawley rats. The female Dark Agouti lacks CYP2D1, the cytochrome P450 enzyme which catalyses the oxidative O-demethylation of dextromethorphan to dextrorphan. No differences were observed in either potency or time to peak effect for dextromethorphan to induce stereotyped behavior in the rat strains, suggesting that the affinity of dextromethorphan for NMDA receptors adequately accounts for its ability to induce stereotyped behavior. Female Dark Agouti rats were, however, more sensitive to the effects of dextrorphan, which may reflect differences in the ability of this strain to metabolize dextrorphan. We find no evidence to suggest that dextromethorphan produces a behavioral syndrome in rats that is distinct from that induced by dextrorphan. The commonality between the pharmacologic profiles of these compounds suggests that the abuse potential of dextromethorphan containing antitussive preparations is related to the non competitive NMDA antagonist activity of dextromethorphan and its metabolites. PMID- 9860114 TI - Excitotoxic lesions of the mediodorsal thalamic nucleus attenuate intravenous cocaine self-administration. AB - The present experiments investigated the effects of excitotoxic, axon-sparing lesions of the mediodorsal nucleus of the thalamus (MD) on locomotor activity and i.v. cocaine self-administration. Infusion of quinolinic acid into the MD using a glass micropipette produced well-defined neuronal loss restricted to medial and lateral portions of the MD, sparing adjacent areas such as the lateral habenula and paraventricular thalamic nucleus. MD lesions resulted in delayed habituation to activity cages. In addition, lesioned rats self-administered significantly smaller amounts of cocaine than controls during a 14-day acquisition period, and showed attenuated responding for cocaine doses on the descending limb of the dose effect function. Since typical titrating patterns of responding were maintained in lesioned rats, and responding on the inactive lever did not differ from sham operated animals, these present results indicate an enhanced sensitivity to the reinforcing effects of response-contingent cocaine in rats with excitotoxic lesions of the MD. PMID- 9860113 TI - Opioid enhancement of the discriminative stimulus effects of cocaine: evidence for involvement of mu and delta opioid receptors. AB - Previous research in squirrel monkeys has shown enhancement of the discriminative stimulus effects of cocaine by mu-opioid agonists, but not by the delta agonist BW373U86. To examine further the role of mu and delta receptor stimulation in the ability of opioid drugs to modulate the discriminative stimulus effects of cocaine, the present study assessed the effects of cocaine alone and combined with SNC 80, a selective high-efficacy delta agonist, and fentanyl, a selective high-efficacy mu agonist. Five adult male squirrel monkeys were trained to discriminate i.m. injections of 0.3 mg/kg cocaine from saline under a fixed-ratio 10 schedule of food presentation. Cumulative doses of cocaine (0.03-1.0 mg/kg) engendered dose-related increases in drug-lever responding to a maximum of 100%, with a decrease in response rate observed at 1.0 mg/kg. Cumulative doses of SNC 80 (0.03-1.0 mg/kg) or fentanyl (0.001-0.01 mg/kg) resulted in a maximum of 22% and 48% drug-lever responding, respectively, accompanied by pronounced decreases in response rate. Administration of either SNC 80 (0.1-1.0 mg/kg) or fentanyl (0.001-0.01 mg/kg) prior to cumulative doses of cocaine produced dose-dependent leftward shifts in the cocaine dose-response function. When the selective delta antagonist naltrindole (1.0 mg/kg) was combined with SNC 80 (1.0 mg/kg) or fentanyl (0.01 mg/kg) prior to cumulative doses of cocaine, the leftward shift of the cocaine dose-response function produced by SNC 80 was blocked, whereas the leftward shift produced by fentanyl was not. By contrast, the mu antagonist naltrexone (0.3 mg/kg) blocked the cocaine-enhancing effects of fentanyl, but not of SNC 80. Combinations of SNC 80 (0.03-0.3 mg/kg) with fentanyl (0.001-0.003 mg/kg) resulted in leftward shifts in the cocaine dose-response function that were comparable in magnitude to the shifts in the cocaine dose-response function produced by either drug alone. These results suggest that opioid enhancement of the discriminative stimulus effects of cocaine is mediated independently by delta and mu-receptor mechanisms. PMID- 9860115 TI - Adaptations in the mesoaccumbens dopamine system resulting from repeated administration of dopamine D1 and D2 receptor-selective agonists: relevance to cocaine sensitization. AB - The mesoaccumbens dopamine (DA) system is intricately involved in sensitization to the locomotor stimulant effects of cocaine. Among the adaptations implicated in cocaine sensitization are transient subsensitivity of impulse-regulating DA D2 autoreceptors on ventral tegmental area (VTA) DA neurons leading to hyperactivity of the mesoaccumbens DA pathway, and persistently enhanced DA D1 receptor responses of nucleus accumbens (NAc) neurons. We have tested the hypothesis that both of these adaptations are necessary to produce cocaine sensitization. We injected rats twice daily for 2 weeks with the selective DA D1 class receptor agonist SKF 38393, the DA D2 class receptor agonist quinpirole, or both. We then used single-cell recording procedures to determine possible alterations in VTA DA autoreceptor sensitivity and NAc D1 receptor sensitivity at three withdrawal times: 1 day, 1 week and 1 month. We also tested whether these treatments produced cross-sensitization to cocaine at each withdrawal time. Repeated quinpirole treatment produced a reduction in VTA autoreceptor sensitivity and cross-sensitization to cocaine, but these effects lasted for less than 1 week. Repeated SKF 38393 treatment produced enhanced NAc D1 responses which lasted for 1 week and cross-sensitization to cocaine which was only evident after 1 week of withdrawal. Repeated treatment with the combination of the two agonists transiently down-regulated autoreceptor sensitivity, enhanced and prolonged D1 receptor supersensitivity (lasting 1 month), and produced enduring cross sensitization to cocaine. These results suggest that neuroadaptations within both the VTA and NAc may be necessary for the induction of enduring cocaine sensitization. PMID- 9860116 TI - 5-HT3 receptor over-expression decreases ethanol self administration in transgenic mice. AB - The 5-HT3 receptor is thought to play a role in the reward pathway and the phenomena of drug abuse by modulating dopamine release in the mesolimbic pathway. Studies involving this receptor have been hampered due to the low level of 5-HT3 receptors in the CNS. A 5-HT3 receptor over-expressing mouse was produced to study the role of this receptor in the rewarding properties of drugs of abuse. Over-expression was restricted to the forebrain by controlling gene expression with the Ca2+ calmodulin (CAM) kinase IIalpha promoter. No over-expression was detected in other body organs nor the cerebellum, as measured by ligand binding and Northern analysis. 5-HT3 receptor over-expressing mice drank less alcohol than non-transgenic mice in a two-bottle free choice test. Over-expression of the 5-HT3 receptor in these mice resulted in a decrease in ethanol consumption. These mice should prove useful in testing hypothesis regarding a common reward pathway for drugs of abuse and the role 5-HT3 receptors play in this pathway. PMID- 9860117 TI - The story of the internal carotid artery of mammals: from Galen to sudden infant death syndrome. AB - Some anatomical aspects of the blood supply of the brains of mammals have been examined to illuminate their functions. A fundamental explanation of sudden infant death syndrome (cot death) is suggested following experimental observations. Speculative contributions have been made to comparative physiological ideas concerning mammals of pronograde and erect habitus, their vascular pressure adaptations and temperature management. Neuro- and interventional radiologists may make some significant future applications of these ideas. Of immediate practical interest is the possibility of influencing the well-being of human embryos' neural tube development in utero by a comprehensive study of their temperature environment. PMID- 9860118 TI - MR angiography in internal carotid artery dissection: improvement of diagnosis by selective demonstration of the intramural haematoma. AB - We studied eight consecutive patients with clinical and intra-arterial angiographic (DSA) diagnosis of internal carotid artery (ICA) dissection to analyse the accuracy of MRA in dissections, using a new technique with presaturation pulses. Spin-echo images of the head were followed by three dimensional time-of-flight (TOF) MRA at the site of the dissection, with and without a special caudal saturation pulse in addition to the cranial one. The accuracy of MRA was assessed in 64 segments of 16 ICA, all examined with DSA and MRA. High-signal intramural haematoma in the ICA at the level of the dissection was observed in all patients either on the maximal-intensity projection (MIP) reconstructions or on the partitions with this presaturation pulse technique. MRA had a sensitivity to detect dissected vessels of 100%. Specificity for vessels correctly identified as not having a lesion was also 100%. There was good correlation between DSA and MRA in demonstrating the site of the dissected ICA segment and the degree of stenosis. In only two cases was there overestimation of the degree of stenosis on the MIP reconstructions of the 3-D MRA. PMID- 9860119 TI - MRI in occipital lobe infarcts: classification by involvement of the striate cortex. AB - We reviewed the MRI studies of 25 patients with occipital lobe infarcts to clarify the distribution of infarcts in the posterior cerebral arterial territory, focussing on their relationship to the striate cortex. Visual field defects and MRI findings were also correlated in 16 patients. On coronal and/or sagittal images, the distribution of the infarct and its relationship to the striate cortex were classified. Involvement of the cortex of both upper and lower lips of the calcarine fissure was observed in 10 patients, and involvement of the lower lip alone in 15. The upper cortical lesions were always accompanied by lower cortical lesions. The visual field defects were complete hemianopia in nine patients, superior quadrantanopia in six and hemianopia with a preserved temporal crescent in one. All patients with superior quadrantanopia had involvement of the lower cortex alone; there were no cases of inferior quadrantanopia. The characteristic vascular anatomy, and poor development of the collateral circulation in the lower cortical area, may explain the vulnerability of this area to infarcts. PMID- 9860120 TI - MRI of spinal cord and brain lesions in subacute combined degeneration. AB - Subacute combined degeneration is a rare cause of demyelination of the dorsal and lateral columns of the spinal cord and even more rarely of the pyramidal and spinocerebellar tracts and cerebellum. We present the initial and follow-up MRI appearances in a patient with subacute combined degeneration of the spinal cord, brain stem and cerebellum, due to vitamin B12 deficiency. The lesions in these structures were demonstrated clearly as pathologically high-signal areas on T2 weighted images. These lesions, except those of the brain stem and cerebellum, disappeared 4 months after therapy. MRI 14 months after the patient's discharge on vitamin B12 therapy showed the same picture. PMID- 9860121 TI - Hemorrhagic chondroid chordoma mimicking pituitary apoplexy. AB - We describe a hemorrhagic chondroid chordoma involving the sella turcica with suprasellar extension. The CT and MRI appearances mimiked a hemorrhagic pituitary adenoma. Chondroid chordoma is a variant composed of elements of both chordoma and cartilaginous tissue. An uncommon bone neoplasm, located almost exclusively in the spheno-occipital region, it is usually not considered in the differential diagnosis of a tumor with acute hemorrhage in the sellar region. We discuss the clinical and radiological characteristics which may allow one to differentiate chondroid chordoma from other tumors of this area. PMID- 9860122 TI - MRI demonstration of subarachnoid neurocysticercosis simulating metastatic disease. AB - We present a patient with neurocysticercosis with spinal subarachnoid spread who presented with lower back pain and progressive numbness and weakness of the left leg. MRI of the spine simulated metastasis. MRI of the brain demonstrated a "bunch of grapes" appearance in the basal cisterns, characteristic of cysticercosis. PMID- 9860123 TI - L-2-Hydroxyglutaric aciduria: MRI in seven cases. AB - The MRI findings in 7 patients with L-2-Hydroxyglutaric aciduria (L-2-OHG aciduria) are described and compared with previous neuroradiological reports and the only three published pathological cases. Signal abnormalities involved peripheral subcortical white matter, basal ganglia and dentate nuclei. Cerebellar atrophy was present. Although similar appearances may be seen in other metabolic disorders, the distribution of signal abnormalities in L-2-OHG aciduria is highly characteristic and may suggest the correct diagnosis. PMID- 9860124 TI - Hemimegalencephaly: signal changes suggesting abnormal myelination on MRI. AB - We reviewed the MRI of 17 patients with hemimegalencephaly to investigate abnormal myelination in this condition. On images of seven patients aged 18 months or less, the white matter on the affected side suggested advanced myelination for the age. On T1-weighted images of three patients aged 1 month, the anterior limb of the internal capsule in the affected hemisphere was myelinated, and T1 shortening was not clearly seen in the pre- and postcentral gyri. The cortical grey matter and subcortical white matter was isointense in two patients. Images of two patients aged 4 to 5 months and of five patients aged 8 18 months showed myelination that extended more peripherally in the white matter of the affected hemisphere. PMID- 9860125 TI - Lyme disease in a child presenting with bilateral facial nerve palsy: MRI findings and review of the literature. AB - We report a 7-year-old boy with neuroborreliosis presenting with headache and bilateral facial nerve palsy. MRI demonstrated tentorial and bilateral facial and trigeminal nerve enhancement. PMID- 9860126 TI - Clear-cell meningioma of the cauda equina. AB - Meningiomas are rare tumours in children and lumbar lesions are exceptional. We report a clear-cell meningioma (CCM) of the cauda equina in a 10-year-old girl. The tumour was diagnosed by MRI, showing an enhancing intradural mass extending from L1 to L4. Pathology and immunohistochemical study demonstrated a CCM. The patient had a recurrence 6 months after the operation requiring further surgery. CCM are rare lesions, characterised by abundant cytoplasmic glycogen particles. Complete surgical removal is necessary because, despite their benign histological appearance, CCM are potentially aggressive and may recur, spread locally and even metastasize. PMID- 9860127 TI - Anomalous vessel in the middle ear: the role of CT and MR angiography. AB - An 8-year-old child was examined because of conductive hearing loss with a retrotympanic mass on otoscopy. CT and MR angiography showed a large inferior tympanic artery traversing the hypotympanum and joining a thin, irregular internal carotid artery with a normal middle meningeal artery. These investigations, coupled with knowledge of the embryological development allowed a diagnosis of a complex vascular anomaly in the middle ear and avoided potential surgical complications. PMID- 9860128 TI - Radiology of an atypical fibro-osseous lesion confined to the nasal cavity. AB - Fibro-osseous lesions of the sinonasal region are relatively frequent, but those strictly confined to the nasal cavity are rare. We report an atypical fibro osseous lesion in the nasal cavity and describe its radiological features. The differential diagnosis is discussed. PMID- 9860129 TI - Anatomical and morphological factors correlating with rupture of intracranial aneurysms in patients referred for endovascular treatment. AB - The size of intracranial aneurysms is the only characteristic shown to correlate with their rupture. However, the critical size for rupture has varied considerably among previous accounts and remains a point of controversy. Our goal was to identify statistically significant clinical and morphological factors predictive of the occurrence of rupture and aneurysm size in patients referred for endovascular treatment. We retrospectively recorded the following factors from 74 patients who presented with ruptured (40) or unruptured (34) aneurysms: aneurysm morphology (uni/multilobulated), location (anterior/posterior), maximum diameter, diameter of the neck, and the patient's age and sex. We performed stepwise discriminant, and stepwise and logistic regression analysis to identify factors predicting rupture and the size of the aneurysm at rupture. The mean diameter of the ruptured aneurysms was 11.9+/-6.3 mm, range 3.0-33.0 mm, and that of the unruptured aneurysm 13.5+/-5.8 mm, range 5.0-30 mm. Stepwise discriminant analysis identified aneurysm morphology (P < 0.001) and location in the intracranial circulation (P < 0.001) as statistically significant factors in predicting rupture. Stepwise regression analysis revealed that aneurysm morphology and the size of the neck were predictors of aneurysm size at rupture. PMID- 9860130 TI - Vertebral arteriovenous fistula: endovascular treatment with electrodetachable coils. AB - We report a patient with an iatrogenic vertebro-vertebral fistula responsible for a cerebellar syndrome and a cervical bruit. Endovascular treatment of the fistula was carried out using electrically detachable coils. An angiogram 5 months after treatment showed that the fistula had totally disappeared. PMID- 9860131 TI - Mint 3: a ubiquitous mint isoform that does not bind to munc18-1 or -2. AB - Mint 1 and 2 are proteins that bind to munc18-1, an essential component of the synaptic vesicle fusion machinery, and are detectably expressed only in neurons [Okamoto and Sudhof, J. Biol. Chem. 272, 31459-31464 (1997)]. Mint 1 and 2 are composed of a variable N-terminal region that includes a conserved munc18-1 binding site, and a constant C-terminal region that contains one PTB and two PDZ domains. We have now identified a third mint isoform, mint 3. Similar to mint 1 and 2, the C-terminal half of mint 3 is composed of one PTB domain and two PDZ domains. However, in contrast to mint 1 and 2, mint 3 lacks an N-terminal munc18 binding domain and does not interact with munc18-1 in yeast two-hybrid assays. Mint 3 is ubiquitously expressed in all tissues, with lowest levels in brain and testis whereas mint 1 and 2 appear to be brain-specific. Our data suggest that mints form a diverse family of proteins with specialized neuronal and ubiquitous isoforms. PMID- 9860132 TI - In vitro fusion of tissue-derived endosomes and lysosomes. AB - We investigated the in vitro fusion of different endocytic compartments derived from perfused rat liver, where the cells are assumed to be in their physiological state. Specifically labelled early, late and transcytotic endosomes, as well as lysosomes were tested for their fusion properties. In addition to the expected ATP-dependent fusion between early endosomes, we observed fusion between early and late endosomes with similar efficiency, kinetics and cytosol dependence. Fusion between early endosomes and transcytotic vesicles could not be detected. Prolonged incubation of complementary labelled early endosomes under fusion supporting conditions followed by Percoll gradient centrifugation revealed the occurrence of fusion product at a dense position, indicating fusion events between light and dense compartments. Incubation of membrane preparations containing avidin-labelled endosomes and biotin-dextran-loaded lysosomes resulted in the formation of avidin-biotin complexes, indicating that fusion between early and late endosomes is followed by fusion with lysosomes. This was verified by colocalization of fluorescently labelled endosomes and lysosomes, as assessed by laser scanning microscopy. Endosome fusion, as well as content mixing between endosomes and lysosomes, were dependent on temperature and ATP, and could be inhibited by N-ethylmaleimide (NEM). The NEM-sensitivity was localised on endosomes and in the cytosol, but not on lysosomes. These observations indicate that early and late endosomes of rat liver exhibit a high fusion competence in vitro, promoting not only homotypic, but also heterotypic fusion. PMID- 9860133 TI - Zebrafish vimentin: molecular characterization, assembly properties and developmental expression. AB - To provide a basis for the investigation of the intermediate filament (IF) protein vimentin in one of the most promising experimental vertebrate systems, the zebrafish (Danio rerio), we have isolated a cDNA clone of high sequence identity to and with the characteristic features of human vimentin. Using this clone we produced recombinant zebrafish vimentin and studied its assembly behaviour. Unlike other vimentins, zebrafish vimentin formed unusually thick filaments when assembled at temperatures below 21 degrees C. At 37 degrees C few filaments were observed, which often also terminated in aggregated masses, indicating that its assembly was severely disturbed at this temperature. Between 21 and 34 degrees C apparently normal IFs were generated. By viscometry, the temperature optimum of assembly was determined to be around 28 degrees C. At this temperature, zebrafish vimentin partially rescued, in mixing experiments, the temperature-dependent assembly defect of trout vimentin. Therefore it is apparently able to "instruct" the misorganized trout vimentin such that it can enter normal IFs. This feature, that assembly is best at the normal body temperature of various species, puts more weight on the assumption that vimentin is vital for some aspects of generating functional adult tissues. Remarkably, like in most other vertebrates, zebrafish vimentin appears to be an abundant factor in the lens and the retina as well as transiently, during development, in various parts of the central and peripheral nervous system. Therefore, promising cell biological investigations may now be performed with cells involved in the generation of the vertebrate eye and brain, and, in particular, the retina. Moreover, the power of genetics of the zebrafish system may be employed to investigate functional properties of vimentin in vivo. PMID- 9860135 TI - Changes in actin cytoskeleton during volume regulation in C6 glial cells. AB - Changes in actin cytoskeleton in the C6 rat glial cell line were studied during decrease or increase (abrupt or gradual) of extracellular osmolality. Actin cytoskeleton was visualized by confocal microscopy after FITC-phalloidin labeling. G-actin, Triton-soluble F-actin and Triton-insoluble F-actin subfractions were determined by gel electrophoresis and scanning, and by DNase I inhibition assays. In control conditions C6 glial cells exhibited well-defined stress fibers and a relatively smooth cortical network. Extracellular anisosmotic changes induced a rapid actin cytoskeletal reorganization, which further progressed and was not reversed upon cell volume recovery. Hypotonic shock caused membrane ruffling and a shift towards polymerized actin, whereas hypertonicity (abrupt or gradual) led to a distinct morphological appearance of abundant short actin microfilaments with, however, no detectable alteration in actin subfractions. When anisosmotic cell volume regulation was prevented, cytoskeleton reorganization depended on the osmotic change and the experimental protocol, but was not related to the absence of volume readjustment. Therefore, although involvement of cytoskeletal alterations in transduction of volume regulatory responses cannot be excluded, it is likely that the observed changes in actin cytoskeleton in C6 glial cells are linked with, but do not initiate, cell volume regulatory processes. PMID- 9860134 TI - Synthesis of proto-oncogene proteins and cyclins depends on intact microfilaments. AB - It is well established that microfilament disintegration by cytochalasin D (CD) as well as latrunculin (LAT)-A and LAT-B causes an inhibition of S phase entry of various nontransformed cell lines. Our experiments extended these observations to human embryonal diploid fibroblasts (Wi-38). To investigate the question whether this stop of DNA synthesis is due to a decline of the synthesis of proteins that are necessary for G1 progression and S phase entry, we examined the expression of two proto-oncogenes (c-fos, c-jun) and three cyclins (D1, E, A) after altering the microfilament system. Disintegration of microfilaments by CD, LAT-A, or LAT-B of asynchronously growing fibroblasts caused a strong dose-dependent and time dependent inhibition of total protein synthesis. Expression of c-jun, cyclins D1, E, and A decreased by about the same percentage as total protein synthesis. The strong induction of total protein synthesis after reactivating serum-starved fibroblasts by adding fetal calf serum was suppressed, when CD or LAT-A were added to the culture medium during this reactivation process. While expression of cyclin E as well as cyclin A decreased by about the same percentage as total protein synthesis, cyclin D1 was more suppressed after microfilament disintegration. After reactivating growth-arrested Wi-38 fibroblasts, cultured in suspension for 12 h, by transferring them to a rigid substratum they could adhere to, total protein synthesis was strongly induced. Again alteration of microfilaments by CD suppressed that increase. The expression of cyclin D1 was slightly more suppressed than total protein synthesis after addition of CD during that reactivation process. Our results suggest that alteration of microfilaments causes a strong decline of total protein synthesis accompanied by a decrease of the expression of proteins that are required for G1 progression and S phase entry. The diminished presence of proteins that are important for cell cycle progression could explain the inhibition of DNA synthesis after microfilament disintegration by various drugs. PMID- 9860136 TI - Ciliated differentiation of rabbit tracheal epithelial cells in vitro. AB - Primary cultures of rabbit tracheal epithelial (RbTE) cells have been performed in two different ways. Quantitative analysis of both proliferative capacities and ciliated differentiation process were carried out using epithelial cell cultures from tracheal explants and from dissociated tracheal epithelial cells in air liquid interface conditions. We show that both alpha- and beta-tubulins from RbTE cells are polyglutamylated and that this posttranslational modification is restricted to cilia axonemes and centrioles of non-ciliated cells. A monoclonal antibody raised against polyglutamylated tubulins was used to quantify the proportion of ciliated cells. Even though epithelial cells from outgrowths obtained by the explant technique highly proliferated during the first days of culture, no ciliated differentiation occurred. On the other hand, using air liquid interface conditions after proliferation of dissociated cells, we could observe and quantify a ciliated cell differentiation in vitro by both Western blot and flow cytometric analysis. The specific detection and quantification of ciliated cells open the way for the biochemical and molecular characterization of centriolar components during ciliated differentiation. PMID- 9860137 TI - Enhanced differentiation of HL-60 leukemia cells to macrophages induced by ciprofibrate. AB - Ciprofibrate, an hypolipidaemic peroxisome proliferator, induced differentiation of HL-60 cells. The effect was greatly potentiated by phorbol 12-myristate 13 acetate at a concentration where neither phorbol ester nor ciprofibrate alone had any effect on these cells. As occurs for HL-60 cell differentiation induced by high phorbol ester concentration, the ciprofibrate-induced phorbol ester dependent differentiation of HL-60 cells proceeded through the monocytic/macrophage pathway and induced the phosphorylation of proteins with similar molecular weights suggesting that increased protein kinase C activity may be involved in the effect. The peroxisome proliferator-activated receptor (PPARalpha) transcription factor is expressed in HL-60 cells, but no changes were observed in its expression upon HL-60 cell differentiation. PMID- 9860138 TI - The role of PDGF-dependent suppression of apoptosis in differentiating 3T3-L1 preadipocytes. AB - In a chemically defined serum-free culture system, platelet-derived growth factor (PDGF) as the only externally applied growth factor, in concert with corticosterone, 3-isobutyl-1-methylxanthine (IBMX) and low insulin (1nM), stimulates adipose conversion of 3T3-L1 preadipocytes. Omission of PDGF during the induction period results in loss of differentiation competence and apoptotic cell death. Induction of apoptosis is shown to be clearly mediated by PDGF withdrawal, since neither corticosterone nor IBMX affect the apoptotic behaviour of 3T3-L1 cells. Cell viability in the absence of the survival factor PDGF could be achieved by application of high insulin (1 microM) or ectopical expression of the anti-apoptotic proto-oncogene Bcl-2. However, PDGF-independent suppression of cell death does not trigger adipose conversion in the presence of corticosterone and IBMX. Therefore, we conclude that suppression of apoptosis per se is not permissive for differentiation of 3T3-L1 preadipocytes and PDGF might exert some additional differentiation-promoting effect(s). PMID- 9860139 TI - Control of the human cell cycle by a bacterial protein, gapstatin. AB - The oral gram-negative bacterium Actinobacillus actinomycetemcomitans is a major pathogen in human periodontal disease. Saline extraction releases a range of surface-associated components from this bacterium, including one which exhibits potent anti-proliferative activity as assessed by its capacity to inhibit DNA synthesis by human and other mammalian cells. Cultures incubated with this bacterial fraction for a prolonged period comprise a high proportion of cells containing a 4n level of DNA. Studies using hydroxyurea-synchronized cultures showed that cells treated with the surface-associated fraction were arrested in the G2 phase of the cell cycle and did not enter mitosis. This G2/M blockade was observed only when the bacterial fraction was added to the cells during early S phase. Our data also suggest that the active bacterial component binds to surface receptors expressed by the human cells and may act by a novel mechanism which involves down-regulation of cyclin B1 expression. The anti-proliferative activity of the bacterial fraction, purified by a combination of ammonium sulphate precipitation, HPLC anion exchange and gel filtration, has been shown to be an 8 kDa protein, which we have called gapstatin. Purified gapstatin was shown to be responsible for the the inhibitory effects of the surface-associated fraction on mammalian cells. PMID- 9860140 TI - Euchromatin megabase cleavages and conjoint apoptotic-autophagic death expression with nucleolar ball-and-socket joint dislocations in human Chang liver cells arrested in S-phase by etoposide. AB - Etoposide induced a megabase (Mb) fragmentation pattern identical with that from genomic digestion by NotI restriction endonuclease which specifically cleaves CpG islands in euchromatin domains. Redigestion by NotI produced no change, suggesting cleavage in the same or closely related sites in euchromatin domains. Preferential euchromatin cleavage was further suggested by harvested metaphase chromosomes showing self-inflicted resolution of light G-bandings (R-bandings), the euchromatin domains. Autodegeneration following Mb euchromatin fragmentations was shown by their degradation into 200 bp ladders, and expressions of apoptotic and "non-apoptotic" active death morphologies that were also seen conjointly in the same cell. The endstage further showed heterochromatin masses anchored to the nucleolus by novel ball-and-socket joints where dislocations occurred with nuclear leakage. PMID- 9860141 TI - Replication of 5 S ribosomal genes precedes the appearance of early nuclear replication complexes. AB - The present work shows that replication of the 5 S ribosomal genes differs in time and 3'deoxyadenosine sensitivity from replication of other nuclear genes, in Allium cepa L. root meristems. Fluorescence in situ hybridization with the pTa794 DNA probe which contains a complete 410 bp 5 S gene from Triticum aestivum allowed to detect four clusters of 5 S genes in these diploid cells (2n = 16), two of them in the short arm of the smallest metacentric chromosomal pair 7. Replication of the 5 S ribosomal genes occurred very early in interphase, as discerned by their resolution as doubled spots only two hours after interphase was initiated in synchronous binucleate cells. Codetection of nuclear replication (by immunodetection of 5-bromo-2'-deoxyuridine incorporation) showed that the replication of the 5 S ribosomal genes occurred before any incorporation of 5 bromo-2'deoxyuridine could be detected in the nuclei. The earliest Br-DNA detected in these cells followed a radial pattern from different foci apparently dispersed along some chromosomal arms. These structures seem to represent early replication complexes, as a result of the displacement of multiple DNA forks from the foci known as pre-replication complexes where the replication machinery of the earliest replicating genes assembles. No consistent positional correlation existed between the formation of the early replication complexes and the already replicated 5 S ribosomal clusters. Finally, nuclear replication but not that of the 5 S genes was prevented by 3'deoxyadenosine, and the earliest replicating 5 S ribosomal gene cluster differed in both sister nuclei resulting from the segregation of one single chromosome in anaphase. PMID- 9860142 TI - Identification of a phospholipase C beta subtype in rat taste cells. AB - From rat circumvallate papillae a novel phospholipase C (PLC) subtype has been cloned and identified as most closely related to human PLC beta2. The corresponding mRNA was only detected in sensory lingual tissue but not in non taste lingual tissue or any other tissues examined by Northern blot analysis. In situ hybridization revealed that a subset of taste receptor cells of circumvallate papillae was specifically labeled. A functional involvement of this PLC beta subtype in taste signal transduction emerged from biochemical analysis monitoring the second messenger response in circumvallate preparations induced by denatonium benzoate. This bitter agent elicited a rapid and transient increase of the inositol 1,4,5-trisphosphate level; this response was blocked by U73122, a potent inhibitor of PLC, and by PLC beta2-specific antibodies. These data indicate that a phospholipase C beta2 isoform mediates a denatonium benzoate induced second messenger response of taste sensory cells in the circumvallate papillae. PMID- 9860143 TI - The progress of COST B1--a European collaboration on the study of interindividual differences in drug disposition and action. Organization for European Co operation in the field of Science and Technical Research. PMID- 9860144 TI - Adverse effects profile of the herbal antidepressant St. John's wort (Hypericum perforatum L.). AB - This paper provides a systematic review of adverse drug reactions (ADRs) associated with the use of extracts of the herb St. John's wort (Hypericum perfbratum L.) for the treatment of mild to moderate depression. METHODS: Searches of four computerized literature databases were performed for records of (ADRs). Manufacturers of hypericum products, the international drug monitoring centre of the World Health Organization (WHO) and the national drug safety monitoring bodies of Germany and the United Kingdom were also contacted for information. RESULTS: Information on (ADRs) originates from case reports, clinical trials, post-marketing surveillance and drug monitoring studies. Collectively, the data suggest that hypericum is well tolerated, with an incidence of adverse reactions similar to that of placebo. The most common adverse effects are gastrointestinal symptoms, dizziness/confusion and tiredness/sedation. A potential serious adverse effect is photosensitivity, but this appears to occur extremely rarely. CONCLUSIONS: Hypericum has an encouraging safety profile. However, as most of the current data originate from short-term investigations, more long-term studies are desirable. PMID- 9860145 TI - Haemodynamic and renal effects of felodipine in young and elderly subjects. AB - OBJECTIVE: To study the influence of age on renal and haemodynamic effects of the calcium antagonist felodipine. METHODS: Eight young (mean age 27 years) and eight elderly (mean age 75 years) healthy normotensive subjects were given felodipine intravenously for 120 min aiming at close to therapeutic plasma level concentration. Renal blood flow (RBF) and renal vascular resistance (RVR) was estimated from para-aminohippuric acid (PAH) clearance 51CrEDTA clearance was used to measure glomerular filtration rate (GFR) and used in the calculations of fractional excretion (FE) of electrolytes. Impedance cardiography was performed to assess stroke volume and for the calculation of cardiac output and ejection fraction. RESULTS: At the end of felodipine infusion, the concentration of felodipine was on average 10.0 nmol x l(-1) in young and 12.0 nmol x l(-1) in elderly subjects (NS). During felodipine infusion blood pressure (BP) decreased from 138/76 to 120/68 in elderly subjects. The BP in young subjects was 126/74 at basal and 125/70 after infusion of felodipine. The systemic and renal vascular resistance decreased to a similar extent in young and elderly subjects after felodipine infusion. Felodipine caused a decrease in systemic vascular resistance from 25.6 to 23.3 in elderly and from 23.8 to 21.8 in the young subjects. Mean values for RVR at baseline and during infusion of felodipine were significantly higher in the elderly (10.1-15.1) than in the young subjects (5.4-6.7). Felodipine reduced RVR by 10% in the young and by 12% in the elderly at the end of infusion. The young subjects had 31% higher GFR than the elderly subjects at the start of infusion. Felodipine infusion did not affect GFR. There were no effects on stroke volume and ejection fraction. An initial natriuretic effect was found after infusion of felodipine in the young subjects. The fractional excretion of all electrolytes tended to increase after both felodipine and placebo, more in the elderly than in the young subjects. CONCLUSION: The effects of felodipine on central and renal haemodynamics previously observed in young and middle-aged subjects also seem to exist in the elderly. Volume expansion seems to increase the excretion of electrolytes more in elderly than in young people, and therefore the effect of felodipine on natriuresis is more evident in young subjects. PMID- 9860146 TI - Antioxidative action of the novel calcium channel antagonist mibefradil on low density lipoproteins. AB - OBJECTIVE: Mibefradil is a novel calcium channel antagonist that selectively blocks T-channels. It acts to reduce hypertension, is cardioprotective and reduces ischemic episodes. Oxidative modification of low-density lipoproteins (LDL) is well known to contribute to coronary atherosclerosis and we therefore investigated to see whether mibefradil had antioxidative action on LDL. METHODS: Human LDL were isolated by ultracentrifugation. In vitro oxidation of LDL (0.1 micromol x l(-1) protein) in the presence of various concentrations of mibefradil was initiated by 3.2 micromol x l(-1) copper ions. The kinetics of formation of conjugated dienes was followed photometrically. Malondialdehyde and lipoperoxides were determined at maximum oxidation. LDL (0.3 micromol x l(-1)) were also pre incubated with mibefradil (120 micromol x l(-1)). Excessive mibefradil was separated by column technique. The resultant LDL were oxidized using copper ions or (AAPH) 2,2'-azobis(2-amidinopropane) hydrochloride. RESULTS: The presence of mibefradil in the concentration range from 10 to 200 micromol x l(-1) had dose dependent effects. These were protection of LDL against oxidation measured as prolongation of the lagtime up to 250%, and reduction in the formation of malondialdehyde down to 65% and of lipoperoxides to 20%. Pre-incubation of LDL with mibefradil prolonged the lagtime of Cu-mediated oxidation up to 132% and of AAPH-mediated oxidation up to 138%. CONCLUSION: In addition to the T-channel blocking and antiproliferative effects, our results provide arguments for a protective role of mibefradil (10-200 micromol x l(-1)) on LDL against in vitro oxidation. This was shown with three independent parameters (lagtime, malondialdehyde and lipoperoxides) and in different oxidation models. PMID- 9860147 TI - Buccal absorption of enalapril and lisinopril. AB - OBJECTIVE: The buccal absorption of captopril does not exhibit the classical pH/partition hypothesis, suggesting that mechanisms other than passive diffusion are involved in its absorption; animal studies have suggested that a peptide carrier-mediated transport system may be responsible for its absorption. The present study evaluated the effects of pH on octanol partitioning, and on the buccal absorption of enalapril and lisinopril, using in vitro techniques and buccal partitioning in human volunteer subjects. METHODS: The partitioning of enalapril and lisinopril into n-octanol was examined over the pH range of 3 9 at room temperature. RESULTS: Enalapril exhibited maximal partitioning into the organic phase at pH 4 5; minimal partitioning was recorded at pH values 8 and 9. The partitioning of lisinopril into n-octanol was found to be maximal at pH 9 and minimal at pH 3. Using the buccal absorption technique, the partitioning of enalapril and lisinopril (0.5 mg), was examined in six healthy male volunteers from buffered solutions (pH 3, 4, 5, 6, 7, 8 and 9). In the case of enalapril, lowest buccal partitioning occurred at pH 3, 8 and 9, while maximal partitioning occurred at pH 5; absorption of lisinopril was not extensive at any pH, but was greatest at pH 6. These results, in addition to the n-octanol partition coefficients, indicated that enalapril obeyed the normal lipid partition hypothesis with respect to buccal absorption. The buccal absorption of lisinopril also obeyed the lipid partition hypothesis over the pH range 3-7. These findings are in direct contrast to those for captopril. The buccal partitioning experiments were repeated at the maximal pH for absorption for each angiotensin converting enzyme (ACE) inhibitor, but with the addition of cephradine (0.05 mmol x l(-1)). CONCLUSION: The data indicated that the presence of this peptide transport inhibitor had no effect on the buccal absorption of enalapril (0.06 mmol x l(-1)) and lisinopril (0.057 mmol x l(-1)), which suggests that both drugs do not share a common buccal absorption pathway with cephradine. PMID- 9860148 TI - Lack of bioequivalence of a generic mefloquine tablet with the standard product. AB - OBJECTIVE: To assess the bioequivalence between a generic tablet of mefloquine (Mephaquin = M1) with the reference tablet (Lariam = M2) in healthy volunteers. METHODS: This open label, randomized two-way crossover study was performed in a single centre. Following an overnight fast, eighteen healthy volunteers received a single oral dose of 750 mg mefloquine either in the form of three M1 lactabs or three M2 tablets. Serial blood samples were collected up to 8 weeks after drug administration. Plasma samples were analysed for mefloquine and its carboxylic acid metabolite using liquid chromatography and subsequent tandem mass spectrometry (LC-MS/MS). The pharmacokinetic parameters of mefloquine and its metabolite were estimated by non-compartmental methods. RESULTS: The pharmacokinetics of mefloquine after administration of M1 and M2 tablets were significantly different as reflected by the respective mean values of maximum plasma concentration (Cmax 656 vs 1018 ng x ml(-1)), time to reach maximum concentration (tmax 46 vs 13 h) and area under the plasma concentration-time curve (AUC0-->infinity 338 vs 432 microg x h x ml(-1)). No significant differences existed between the elimination half-lives of the two formulations (394 vs 396 h). The relative bioavailability (M1 vs M2) was 0.78 and ranged from 0.38 to 1.37. Bioequivalence could not be demonstrated for log-transformed data of AUC0-->infinity or AUC0-->last within a predefined range of 80-125% and for Cmax within a range of 70-143%. CONCLUSIONS: The observed differences in Cmax, tmax and AUC are consistent with a slower rate and lower extent of mefloquine absorption after administration of M1. Statistical evaluation of these kinetic data showed that the M1 tablet is not bioequivalent to the M2 tablet. Clinical consequences of this finding cannot be excluded. PMID- 9860149 TI - Vancomycin dosing in morbidly obese patients. AB - OBJECTIVES AND METHODS: Vancomycin hydrochloride dosing requirements in morbidly obese patients with normal renal function were computed to determine the dose of vancomycin necessary to achieve target steady-state peak and trough concentrations and compared with a normal weight population. RESULTS: Morbidly obese patients [total body weight (TBW) 165 kg, ideal body weight (IBW) 63 kg] required 31.2 mg x kg(-1) x d(-1) TBW or 81.9 mg x kg(-1) x d(-1) IBW to achieve the target concentrations. Normal weight patients (TBW 68.6 kg) required 27.8 mg x kg(-1) x d(-1) to achieve the same concentrations. Because of altered kinetic parameters in the morbidly obese patients (obese: t1/2 = 3.3 h, V = 52 L, CL = 197 ml x min(-1); normal: t1/2=7.2 h, V=46 L, CL=77 ml x min(-1), 20 of 24 patients required q8h dosing (1938 mg q8h) compared with q12h dosing (954 mg q12h) in all normal weight patients in order to avoid trough concentrations that were too low for prolonged periods. There was a good correlation between TBW and CL, but only fair correlation between TBW and V. CONCLUSION: Doses required to achieve desired vancomycin concentrations are similar in morbidly obese and normal weight patients when TBW is used as a dosing weight for the obese (approximately 30 mg x kg(-1) x d(-1)). Shorter dosage intervals may be needed when dosing morbidly obese patients so that steady-state trough concentrations remain above 5 microg x ml(-1) in this population. Because of the large amount of variation in required doses, vancomycin serum concentrations should be obtained in morbidly obese patients to ensure that adequate doses are being administered. Dosage requirements for morbidly obese patients with renal dysfunction require further study. PMID- 9860150 TI - Distribution and metabolism of N(G)-nitro-L-arginine methyl ester in patients with septic shock. AB - OBJECTIVE: The pharmacokinetics of N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthesis, was investigated in patients with septic shock. METHODS: Blood was sampled at intervals before, during and after 12 h infusion of L-NAME 1 mg x kg(-1) x h(-1) in nine septic shock patients for determination of plasma concentrations by high-performance liquid chromatography (HPLC). In three patients the renal clearance of the drug was determined. RESULTS: Incubation of L-NAME with plasma and blood in vitro revealed hydrolysis to N(G)-nitro-L-arginine (L-NOARG), the active inhibitor of NO synthesis. L-NOARG did not undergo further degradation. Continuous intravenous infusion of 1 mg x kg(-1) x h(-1) of L-NAME for 12 h in patients with septic shock increased blood pressure and resulted in increasing plasma concentrations of L-NOARG (Cmax 6.2 microg x ml(-1) at 12 h) whereas L-NAME concentrations reached a plateau within 1.5 h (Cmax 1.0 microg x ml(-1)). After the infusion was stopped L-NAME disappeared from the plasma rapidly (half-life 19.2 min) whereas L-NOARG concentration declined slowly (half-life 22.9 h). The calculated volume of distribution for L-NAME was 0.451 x kg(-1) body weight and 1.961 x kg(-1) for L NOARG. The renal clearance for L-NOARG was 3.5% of total body clearance for L NOARG, whereas L-NAME could not be detected in urine. CONCLUSION: We conclude that vasoconstriction with L-NAME in septic patients may result from hydrolysis to L-NOARG, the active inhibitor of NO synthesis. The long plasma half-life and large volume of distribution for L-NOARG suggests extensive distribution to extravascular tissues. Since renal excretion is minimal, elimination of the metabolite L-NOARG follows other pathways. PMID- 9860151 TI - Sparfloxacin pharmacokinetics in healthy volunteers: the influence of acidification and alkalinization. AB - OBJECTIVE: To investigate the effect of acidification and alkalinization on the pharmacokinetics of sparfloxacin in healthy subjects. METHODS: A single 200-mg oral dose of sparfloxacin was given to nine healthy Japanese volunteers on three separate occasions under different conditions of urinary pH. Acidic and alkaline conditions were achieved by repeated oral doses of ammonium chloride and sodium bicarbonate, respectively. The concentrations of sparfloxacin and its metabolite in plasma and urine were determined by high-performance liquid chromatography assays. RESULTS: The difference between treatments for Cmax, AUCinfinity, and CL x f(-1) were found to be significant. The relative bioavailability of sparfloxacin was 84.4% and 122.3% after ammonium chloride and sodium bicarbonate treatments, respectively. The amount of unchanged sparfloxacin in urine samples collected 0-48 h after sparfloxacin administration represented 10.1% of the dose in the control, 14.3% of the dose in urine acidification and 8.4% of the dose with alkalinization of urine. Renal clearance was found to depend on urinary pH. However, the plasma elimination and the metabolism of sparfloxacin were not significantly altered by acidification or alkalinization of the urine. CONCLUSION: The urinary pH dependence of the renal clearance of sparfloxacin will be of minor clinical importance with regard to the low contribution of renal excretion to the overall elimination of sparfloxacin. On the other hand, the alteration in the environmental pH in the gastrointestinal tract, produced by the concomitant ingestion of ammonium chloride or sodium bicarbonate, influences the absorption and bioavailability of sparfloxacin. This effect is likely to be clinically significant. PMID- 9860152 TI - A pharmacokinetic interaction between carbamazepine and olanzapine: observations on possible mechanism. AB - OBJECTIVE: Olanzapine is a novel antipsychotic, which is effective against both the positive and negative symptoms of schizophrenia and causes fewer extrapyramidal adverse effects than conventional antipsychotics. The purpose of the present study was to assess the potential for a pharmacokinetic interaction between olanzapine and carbamazepine, since these agents are likely to be used concomitantly in the treatment of manic psychotic disorder. METHOD: The pharmacokinetics of two single therapeutic doses of olanzapine were determined in 11 healthy volunteers. The first dose of olanzapine (10 mg) was taken alone and the second dose (10 mg) after 2 weeks of treatment with carbamazepine (200 mg BID). Measurement of urinary 6beta-hydroxycortisol/cortisol excretion was used as an endogenous marker to confirm that induction of CYP3A4 by carbamazepine had occurred. RESULTS: The dose of olanzapine given after a 2-week pretreatment with carbamazepine was cleared more rapidly than olanzapine given alone. Olanzapine pharmacokinetic values for Cmax and AUC were significantly lower after the second dose, the elimination half-life was significantly shorter, and the clearance and volume of distribution were significantly increased. CONCLUSION: Carbamazepine has been shown to induce several P450 cytochromes including CYP3A4 and CYP1A2. Since CYP1A2 plays a role in the metabolic clearance of olanzapine, the interaction may be attributed to induction of CYP1A2 by carbamazepine, leading to increased first-pass and systemic metabolism of olanzapine. The interaction is not considered to be of clinical significance because olanzapine has a wide therapeutic index, and the changes in plasma concentration of olanzapine are within the fourfold variation that occurs without concern for safety in a patient population. PMID- 9860153 TI - Patterns of regular drug use in Spanish childbearing women: changes elicited by pregnancy. AB - OBJECTIVE: To determine drug use in Spanish women before pregnancy and from conception to the awareness of pregnancy (early period of pregnancy, EPP), as well as to analyse attitudinal changes when pregnancy was planned or known. METHODS: Trained gynaecologists used a structured questionnaire to collect demographic and obstetric characteristics, histories of regular drug taking before pregnancy, attitudes towards drug taking during pregnancy and current drug use in the EPP. Women were interviewed at their first antenatal visit during the first trimester. RESULTS: Two hundred and seventy-two women were included (mean age 29.3 years and 66.3 days of gestation). Before pregnancy, 24% regularly took drugs, 70% of them more than twice a week; a significantly higher frequency was found in those receiving public antenatal care and in those who had had less education. In 39% of women, awareness of pregnancy did not change their attitudes towards regular drug use. Among those who decided to suppress their regular drug intake, 58% did so when their pregnancy was confirmed and 42% when they planned it. In women who planned their pregnancy, 30.1% stopped when they tried to become pregnant. However, 62% of all women took drugs during the EPP. In private antenatal care significantly more drugs were taken per patient. By the 168 women 278 compounds were consumed during the EPP: 40% were analgesics (mainly paracetamol and acetylsalicylic acid) and 25% were digestive and metabolic drugs (mainly antacids and laxatives). Drugs were often used more than twice a week, particularly in women receiving public antenatal care and in those who had had less education. CONCLUSION: Drug taking is common in Spanish women of childbearing age, and many of those in our study did not decide to stop during the EPP. Few women avoid drugs when planning a pregnancy. Therefore, gynaecologists must advise against drug taking in patients who wish to become pregnant and suggest that unnecessary drug use be avoided when the pregnancy is already diagnosed. PMID- 9860154 TI - Intra-urban variation of antibiotic utilization in children: influence of socio economic factors. AB - OBJECTIVE: The aim of this study was to investigate the intra-urban variation of antibiotic utilization in children in Malmo and to evaluate the influence of socio-economic factors on this variation. METHODS: In an ecological analysis, the variations in antibiotic utilization in children, expressed as defined daily dose (DDD) or as the number of prescriptions per 1000 inhabitants per day, were compared with variations in socio-economic and demographic factors in the 17 administrative districts of the Swedish city of Malmo (235000 inhabitants). RESULTS: There were large between-area differences in antibiotic utilization, especially in children aged 0-6 years. Socio-economic factors reflecting a privileged situation correlated positively with antibiotic utilization. Thus, in districts with a high median family income and a high employment rate, the utilization of antibiotics was higher than in other districts. Conversely, in districts with a high proportion of blue-collar workers, people with foreign backgrounds and recipients of social benefit, antibiotic utilization was comparatively low. In contrast, the utilization of penicillin V relative to other antibiotics showed an opposite pattern, including positive correlations with the proportion of social benefit, immigrants and blue-collar workers and a negative correlation with employment rate. Conversely, the utilization of macrolides in relation to other antibiotics in children aged 0-6 years was highest in districts inhabited by those who were socio-economically privileged. INTERPRETATION: The findings suggest that utilization of antibiotics in children may vary considerably within a city, that it may increase with the degree of parental affluence, and that such affluence may reduce the utilization of penicillin V relative to other antibiotics. PMID- 9860156 TI - Are adverse drug reactions attributed to fluvoxamine caused by concomitant intake of caffeine? PMID- 9860155 TI - Trends in aspirin, paracetamol and non-steroidal anti-inflammatory drug use in children between 1981 and 1992 in France. AB - OBJECTIVE: Antipyretic/analgesic drugs (AADs) are among the most commonly used drugs in children. Their efficacy and adverse effects have often been debated and new AADs have been introduced over the past few years. The aim of this study was to assess the characteristics of the use of AADs in children in France, and their trends. METHODS: Two surveys on household health care consumption were undertaken in France, in 1981 and in 1992. They included 5060 and 4841 children, respectively. The AADs studied were aspirin, paracetamol and non-steroidal anti inflammatory drugs (NSAIDs). RESULTS: The proportion of children exposed to AADs increased significantly between 1981 and 1992 (+28% in 11 years). Among them, the percentage of subjects treated with aspirin decreased (-27%). In contrast, the percentage increased for paracetamol (+ 19%) and for NSAIDs (+179%). Aspirin was the AAD most used in 1981 (57.4%) and it was replaced by paracetamol in 1992 (71.6%). Nasopharyngitis was the main reason for AAD prescription under the age of 11 years; for older children it was influenza-like syndrome, irrespective of the study year. A change in AAD choice occurred in nasopharyngitis, acute bronchitis and influenza-like syndrome irrespective of the age group, and in otitis/sinusitis between 4 and 10 years. In all these cases aspirin prescription decreased, in contrast with paracetamol and NSAIDs. Self-medication of AAD was uncommon (8.3% for aspirin and 10.3% for paracetamol in 1992) and decreased (-29% and -33%). It was used principally for nasopharyngitis, influenza-like syndrome and pain. CONCLUSION: The consumption of AADs in children is high and is increasing. Paracetamol and NSAIDs tend to replace aspirin prescription in children and physicians have played the main role in this change. PMID- 9860157 TI - Drug information services: initial experiences in Dresden. PMID- 9860158 TI - Exposure-response relationship in the hand-arm vibration syndrome: an overview of current epidemiology research. AB - The complex of vascular, neurologic, and osteoarticular disorders occurring in the upper limbs of vibration-exposed workers is called hand-arm vibration syndrome. There is epidemiologic evidence for an increased occurrence of peripheral sensorineural disorders in occupational groups working with vibrating tools. An excess risk for wrist osteoarthrosis and for elbow arthrosis and osteophytosis has been reported in workers exposed to shocks and low-frequency vibration of high magnitude from percussive tools. However, there are too few epidemiology data to enable reliable conclusions to be drawn about exposure response relationships for both sensorineural disturbances and bone and joint disorders caused by hand-transmitted vibration. Cross-sectional and longitudinal epidemiology studies have shown that occupational exposure to hand-transmitted vibration from a great variety of hand-held tools is significantly associated with an increased occurrence of digital vasospastic disorders called vibration induced white finger (VWF). The proposal of an exposure-response relationship for VWF has been included in an annex to the international standard ISO 5349. The findings of several epidemiology studies have shown a poor agreement between the risk for VWF observed in various occupational groups and that predicted by the ISO 5349 model. Both overestimation and underestimation of the occurrence of VWF have been reported by investigators. It has been argued that the current ISO frequency-weighting curve for hand-transmitted vibration may be inappropriate for the assessment of vibration-induced adverse vascular effects. Alternative exposure-response relationships for VWF have been suggested in recent epidemiology studies. The epidemiology data used to construct current exposure response relationships for vibration-induced injuries are primarily derived from cross-sectional studies. Future epidemiology research should be based on prospective cohort studies because the design characteristics of such studies permit the study of cause-effect relationships and the formulation of etiologic hypotheses. PMID- 9860159 TI - Rate pressure product and oxygen saturation in tourists at approximately 3000 m above sea level. AB - OBJECTIVES: Using modern transportation technology, many travelers easily access moderate altitudes of approximately 3000 m above sea level. In the present study the effects of this altitude on cardiovascular parameters were studied among office workers dwelling at sea level. METHODS: Heart rate, blood pressure, arterial oxygen saturation (SpO2), and electrocardiography were monitored before and after Master's double-step exercise at 2700 and 3700 m. The test consisted of stepping onto and off of two 23-cm steps for 3 min at a predefined rate. RESULTS: The resting values recorded for the heart rate and mean blood pressure at 2700 and 3700 m did not statistically significantly differ from those noted at sea level. However, the increases in these values after exercise were significantly greater at high altitude. The rate pressure product more than doubled after exercise at 3700 m. Electrocardiographic abnormalities were observed in some cases. The postexercise blood lactate concentration was significantly higher at 3700 m than at sea level or at 2700 m, suggesting that the oxygen supply-demand relationship was not balanced at this altitude. Furthermore, exercise provoked an acute reduction in SpO2 at 2700 and 3700 m but showed no effect at sea level. CONCLUSION: These observations suggest that the oxygenation status of the heart might be at risk in many travelers and workers during and after exercise load at an altitude of approximately 3000 m. PMID- 9860160 TI - Nasal symptoms and indices of nasal inflammation in flour-dust-exposed bakers. AB - OBJECTIVE: To analyze whether indices of nasal airway inflammation in bakers were related to nasal symptoms and exposure to airborne flour dust. METHODS: A cross sectional study was performed in 12 currently flour-exposed bakers. They were examined by nasal lavage (NAL), visual inspection, a test of mucociliary clearance, and nasal peak expiratory flow (nasal PEF). NAL fluid was analyzed according to the inflammatory markers eosinophil cationic protein (ECP), indicating eosinophilic activity; myeloperoxidase (MPO), indicating active neutrophils; hyaluronic acid (HA) from active fibroblasts; tryptase, indicating activation of mast cells; and albumin, indicating plasma exudation. The bakers were also questioned about respiratory symptoms and working history. Their current and cumulative exposure to inhalable flour dust was estimated after exposure measurements and information about earlier work tasks. Office workers (n=16) without occupational exposure to dust or any other known nasal irritant or sensitizer served as controls. RESULTS: Personal inhalable dust measurements among the bakers working as dough makers or bread formers ranged from 1.0 to 3.8 mg/m3. Of the 12 bakers, 10 reported at least 1 nasal symptom (crusts, blockage, or a runny nose), a proportion significantly greater than that of the controls (P=0.009). Bakers with nasal symptoms had higher concentrations of markers of inflammation in their NALs as compared with nonsymptomatic bakers. The difference was significant for MPO (P=0.02) and HA (P=0.04) in relation to a runny nose. Tryptase was detected in only one NAL of the bakers. There was a positive correlation between the cumulative dose of inhalable flour dust and concentrations of MPO and HA in NAL. Two bakers were sensitized to wheat; they had the highest NAL concentrations of inflammatory markers. CONCLUSIONS: Our results indicate that flour dust exposure in bakers at levels below the current occupational exposure limit causes nasal mucosal inflammation, which, in turn, is related to nasal symptoms. We propose that the inflammation may be nonallergic, characterized by activation of neutrophils and fibroblasts. PMID- 9860161 TI - Exposure to polycyclic aromatic hydrocarbons in occupational versus urban environmental air. AB - OBJECTIVES: To evaluate the balance between occupational and environmental exposure to suspended particulate matter (SPM) and polycyclic aromatic hydrocarbons (PAHs), comparison measurements were performed in a coal-fired power plant and the urban atmosphere from the town nearby. METHODS: The analysis of SPM for PAH content was done according to a high-performance liquid chromatography (HPLC)-based method. The microscopic assessment was performed using scanning electron microscopy (SEM) by silver coverage of the samples derived by air filter. RESULTS: Contrary to expectations, the results showed low levels of particle-bound PAHs in the occupational environment (< 1 ng benzo(a)pyrene/m3 air) and high levels in urban air (range 80-1250 ng benzo(a)pyrene/m3). The SPM collected from the power plant exhibited non-respirable characteristics (particles larger than 10 microm), whereas urban SPM almost exclusively contained respirable airborne particles (<3 microm). CONCLUSIONS: The PAH burden, combined with the enhanced probability of respiratory absorption, confers a much greater hazard potential to the urban SPM. Under these conditions, in areas or countries in which old technologies remain in use, occupational exposure to SPM containing PAHs might represent a severe underestimation of the total risk as it does not take into account the background air pollution. PMID- 9860162 TI - A LISREL analysis of work-related risk factors and health complaints in the nursing profession. AB - OBJECTIVES: To fit a model which describes the complex interaction between various work-related factors and both musculoskeletal and psychosomatic complaints of nurses simultaneously and to compare the results with those of two LISREL models, in which the health outcome variables are analyzed separately in terms of their correlation with the same set of work-related factors. MATERIALS AND METHODS: Nurses (n=718) working in four Dutch nursing homes completed a questionnaire about work-related factors and musculoskeletal and psychosomatic complaints (response 81%). LISREL was used to describe and analyse the models. RESULTS: The full model had a reasonable overall fit. The strongest (and statistically significant) correlations were found between work postures and musculoskeletal complaints and between psychosocial work demands and psychosomatic complaints. In the two single models, health outcomes showed statistically significant correlations with the same work-related factors as they did in the full model. The full model could be slightly improved when the variable somatized complaints (e.g., dizzyness, headache) was related both to musculoskeletal complaints and to psychosomatic complaints. CONCLUSIONS: Although a model that simultaneously describes more dependent variables appears to provide a better approximation of the complexity of the actual relationship between work related factors and health effects, the models in which only one health outcome is studied seem to give a proper description of the data at hand. This is an important finding because these single health outcome models resemble the usual analysis strategy by means of standard regression analysis. The adapted version of the full model suggests that musculoskeletal complaints of nurses can partly be understood as non-specific health complaints. PMID- 9860164 TI - Sister chromatid exchange induced by chromium compounds in human lymphocytes. AB - The objective of this study was to study was to compare chromium (Cr), nickel chromium (Ni-Cr), and control groups for sister chromatid exchange (SCE) in lymphocytes to obtain an understanding of the mutagenic effect of Cr(VI) in humans. Subjects totaled 91 persons from the 3 groups, including 14 Cr and 34 Ni Cr electroplating workers and 43 control group members. Results showed that blood and urine Cr concentrations were highest among Cr workers (11.39 microg/l, 14.7 microg/g creatinine), next highest among Ni-Cr workers (5.28 microg/l, 6.2 microg/g creatinine), and lowest among the control group (2.36 microg/l, NA). After adjustment for smoking habits, SCE/cell values were 10.6, 9.4, and 8.3 for Cr workers, Ni-Cr workers, and controls, respectively. A synergetic effect was shown on HFC (high-frequency cells) percentages for Cr workers who also smoked. Odds ratios were 31.78 and 3.66 that Cr and Ni-Cr workers would have higher HFC percentages than the control group, respectively. The authors conclude that SCE in lymphocytes is useful for evaluation of the biological effects of environmental mutagens. PMID- 9860163 TI - Air pollution and mortality in Madrid, Spain: a time-series analysis. AB - OBJECTIVE: To assess the relationship, if any, between air pollutant (sulfur dioxide and total suspended particulate) levels and mortality in the city of Madrid during the period 1986-1992, controlling for weather, season, and influenza epidemics. METHODS: Daily death counts were obtained from the Regional Mortality Registry. Pollution data were supplied by the Municipal Monitoring Network. Time-series analysis methodology was used to assess the link between non accidental as well as circulatory- and respiratory-disease mortality, on the one hand, and mean daily concentrations of SO2 and total suspended particulate (TSP), on the other. Multivariate autoregressive integrated moving-average (ARIMA) models were used to adjust for season, temperature, relative humidity, and influenza. A sensitivity analysis was run to assess the robustness of the estimators. RESULTS: Graphical analysis revealed a linear relationship between mortality and TSP. The relationship was logarithmic in the case of SO2. TSP lagged 1 day and SO2 lagged 3 days with an independent effect on mortality. This relationship was produced without the detection of a minimal threshold in emission values. CONCLUSIONS: These results support the hypothesis of an association between pollution levels and mortality between 1986-1992 in Madrid. Additional measures designed to reduce pollution levels without compromising thermal comfort should be implemented. PMID- 9860165 TI - Benzene in environmental air and human blood. AB - OBJECTIVE: To study the blood benzene levels resulting from environmental and occupational benzene exposure. METHODS: Benzene in venous blood was measured in 243 nonoccupationally exposed subjects ("normal" people) and in 167 workers occupationally exposed to benzene. All exposed workers gave blood samples at the end of the work shift and on the following morning before resuming work. Blood benzene was assayed by gas chromatography (GC)-mass spectrometry. Occupational benzene exposure was monitored by environmental personal samplers and measured by GC analysis. RESULTS: The mean occupational benzene exposure for all 167 workers studied was 186 ng/l (58 ppb; range 5 1535 ng/l, 2-500 ppb). Overall, the mean blood benzene level of all workers was 420 ng/1 at the end of the shift and 287 ng/l on the morning thereafter. The blood benzene levels measured the morning after turned out to be significantly lower (t=3.6; P < 0.0001) than those measured at the end of the shift. The mean blood benzene level of the 243 "normal" subjects was 165 ng/l, which was significantly lower than that measured in the workers on the morning thereafter (t=5.8: P < 0.0000001). The mean blood benzene concentration was significantly higher in smokers than in nonsmokers in both the general population (264 versus 123 ng/l) and in the exposed workers. In the group of nonsmoking workers, whose workplace exposure to benzene was lower than 100 ng/l, blood benzene levels were similar (210-202 ng/l) to those measured in the nonsmoking general population (165 ng/l). End-of-shift blood benzene correlated significantly with environmental exposure (y=0.91x + 251; r=0.581; n=162; P < 0.00001). Finally, there was also a significant correlation between blood benzene measured at the end of the shift and that determined on the morning thereafter (y=0.45x + 109; r=0.572; n=156; P < 0.00001). CONCLUSION: Nonsmoking workers occupationally exposed to benzene at environmental levels lower than 100 ng/l (mean 35 ng/l) and the nonsmoking general population exposed to ubiquitous benzene pollution have similar blood benzene concentrations. This suggests that it is impossible to distinguish between occupational and environmental exposure when the benzene level in the workplace is less than 100 ng/l. PMID- 9860166 TI - 1,2- and 1,4-Cyclohexanediol: major urinary metabolites and biomarkers of exposure to cyclohexane, cyclohexanone, and cyclohexanol in humans. AB - The metabolism and toxicokinetics of cyclohexane (CH) and cyclohexanol (CH-ol), important solvents and chemical intermediates, were studied in volunteers after 8 h periods of inhalation exposure at concentrations of 1010 and 236 mg m(-3), respectively (occupational exposure limits: CH, 1050 mg m(-3); CH-ol, 200 mg m( 3)). Of the dose of absorbed parent compounds, the yields of urinary CH-ol and 1,2- and 1,4-cyclohexanediol (CH-diol) were 0.5%, 23.4%, and 11.3%, respectively, after exposure to CH and 1.1%, 19.1%, and 8.4%, respectively, after exposure to CH-ol as determined by a gas chromatography method involving hydrolysis of glucuronide conjugates. The metabolic patterns of CH and CH-ol were very similar to that of cyclohexanone (CH-one) studied in the laboratory previously. For all three compounds, peak excretion of CH-ol occurred at the end of the exposure period, after which it decayed rapidly. Excretion curves of 1,2- and 1,4-CH-diol reached maximal values within 0-6 h postexposure, with subsequent elimination half-lives being 14-18 h. The rate-limiting step in the elimination of CH compounds from the organism is renal clearance of CH-diols. Determination of CH diols in end-of-shift urine samples is recommended as a useful new method of biomonitoring of CH, CH-ol, and CH-one at the workplace. However, due to accumulation of CH-diols in the body during repeated exposure, quantitative relationships between the exposure and the level of CH-diols have to be adjusted according to the day of sampling during the working week. PMID- 9860167 TI - Changes in the classification of carcinogenic chemicals in the work area. Section III of the German List of MAK and BAT Values. AB - Carcinogenic chemicals in the work area are currently classified into three categories in section III of the German List of MAK and BAT Values (list of values on maximum workplace concentrations and biological tolerance for occupational exposures). This classification is based on qualitative criteria and reflects essentially the weight of evidence available for judging the carcinogenic potential of the chemicals. It is proposed that these categories - IIIA1, IIIA2, IIIB - be retained as Categories 1, 2, and 3, to correspond with European Union regulations. On the basis of our advancing knowledge of reaction mechanisms and the potency of carcinogens, these three categories are supplemented with two additional categories. The essential feature of substances classified in the new categories is that exposure to these chemicals does not contribute significantly to risk of cancer to man, provided that an appropriate exposure limit (MAK value) is observed. Chemicals known to act typically by nongenotoxic mechanisms and for which information is available that allows evaluation of the effects of low-dose exposures, are classified in Category 4. Genotoxic chemicals for which low carcinogenic potency can be expected on the basis of dose-response relationships and toxicokinetics, and for which risk at low doses can be assessed are classified in Category 5. The basis for a better differentiation of carcinogens is discussed, the new categories are defined, and possible criteria for classification are described. Examples for Category 4 (1,4 dioxane) and Category 5 (styrene) are presented. PMID- 9860168 TI - Mechanisms of fibrinogen domains: biomaterial interactions. AB - Spontaneous adsorption of fibrinogen is critical to the pathogenesis of biomaterial-mediated inflammatory responses. However, the mechanism by which adsorbed fibrinogen affects phagocyte responses is still not clear. To investigate the molecular interaction between fibrinogen and biomaterials, fibrinogen fragments (D100 and E50) were generated and used in the present study. The results indicate that biomaterial: D100 interaction is essential to fibrinogen-mediated inflammatory responses, because biomaterials precoated with D100, but not E50, prompt strong inflammatory responses. Furthermore, the results from in vitro studies show that whole molecule fibrinogen and D100 exhibit very similar protein:surface interactions. Specifically: (1) both D100 and fibrinogen have high affinity for biomaterial surfaces; and (2) the retention rates of adsorbed D100 in both in vivo and in vitro environments are as high as that for adsorbed fibrinogen. On the other hand, E50 does bind to biomaterials but with low affinity because, once bound, it is not tightly adherent to the biomaterial surfaces. Taken together, the results suggest that the mechanism of fibrinogen mediated inflammatory responses may involve the following three consecutive events: (1) after contact with blood or tissue fluid, the D domain tends to interact with biomaterial surfaces and is important in the tight binding of fibrinogen to implant surfaces; (2) the biomaterial surface then promotes conformational changes within the D domain, exposing P1 epitope (gamma 190-202, which interacts with phagocyte Mac-1 integrin); and (3) the engagement of Mac-1 integrin with P1 epitope then triggers subsequent phagocyte adherence and reactions. PMID- 9860169 TI - Release from alginate enhances the biological activity of vascular endothelial growth factor. AB - A primary factor which limits engineering tissues of substantial size is the lack of nutrients readily available to transplanted cells. One potential solution to this nutrient limitation is to encourage the rapid development of a vascular network within three-dimensional tissue engineering matrices. Vascular endothelial growth factor (VEGF) has been identified as a potent stimulator of angiogenesis in vivo. Though effective at stimulating endothelial cells to form blood vessels VEGF degrades rapidly. Spherical alginate beads (3.3+/-0.1 mm diameter) were examined as a means of delivering biologically functional VEGF at a controlled rate over extended times. The alginate beads demonstrated the ability to incorporate VEGF with an efficiency between 30 and 67%, depending on the processing conditions, and release it at a constant rate (5%/day) for up to 14 days in vitro. The released VEGF, when assayed for its ability to stimulate endothelial cells in culture, was found not only to be functional but more potent (three to five times) than the same mass of VEGF added directly to the culture medium. The release kinetics of freeze dried VEGF containing alginate beads were also examined and found to be comparable to non-freeze dried samples. PMID- 9860170 TI - Adsorption of albumin, collagen, and fibronectin on the surface of poly(hydroxybutyrate-hydroxyvalerate) (PHB/HV) and of poly (epsilon-caprolactone) (PCL) films modified by an alkaline hydrolysis and of poly(ethylene terephtalate) (PET) track-etched membranes. AB - The effect of alkaline hydrolysis on several surface properties of poly(hydroxybutyrate-hydroxyvalerate) (92/8) (PHB/HV) and poly(epsilon caprolactone) (PCL) films and of poly(ethylene terephtalate) (PET) track-etched membranes have been characterized, as well as the adsorption of three proteins normally encountered by mammalian cells in vivo, namely albumin, collagen, and fibronectin. The water contact angle decreases and the number of -COOH functions accessible to a chemical reaction at the surface of PCL increases with alkaline hydrolysis. Analysis by atomic force microscopy pictures reveals a change in surface morphology. The modifications of surface properties are correlated with a two times increase of the adsorption of three radiolabelled proteins. The hydrolysis results in a slight increase in the water contact angle of one face of the PHB/HV film and a sharp increase in the number of -COOH functions. Important morphology changes are also induced. The adsorption of the radiolabelled proteins is almost 100 times higher on the hydrolyzed polymer than on the native surface. The increase in hydrophilicity of different PET batches correlates to an increase in the number of -COOH functions. Nevertheless, the surface chemical composition and rugosity are constant and no significant difference in the amount of radiolabelled fibronectin adsorbed on the different surfaces is detectable. In conclusion, the effect of hydrolysis on the surface properties of each of the polyesters studied as well as the proteins adsorption on the different surfaces are different. The results strongly support the hypothesis that, in the system studied, parameters other than hydrophilicity influence protein adsorption: the main parameters that might play a role are the total surface area accessible to the proteins, as well as the surface chemical composition. PMID- 9860171 TI - Tissue factor expression by rat osteosarcoma cells adherent to tissue culture polystyrene and selected orthopedic biomaterials. AB - Tissue factor (TF), a transmembrane glycoprotein expressed by numerous cell types, plays a critical role in the initiation of blood coagulation at sites of vascular injury. Activated products of the coagulation cascade may then enhance the inflammatory responses associated with wound healing. In the present investigation the ability of rat osteosarcoma (ROS) cells to express TF activity was examined following their growth on tissue-culture polystyrene (TCPS) and selected orthopedic biomaterials (titanium and zirconium alloys, and stainless steel). ROS cells exhibited significant TF activity as evidenced by the conversion of Factor X to Factor Xa in the presence of TF, Factor VIIa, and Ca2+. Factor Xa concentrations ranged from 1.0 fM per cell at 10 min to 6.0 fM per cell after 60 min. Additionally, ROS cells stimulated with calcium ionophore (A23187) exhibited approximately twice the activity of non-stimulated cells when grown on TCPS but not on the metallic substrates. ROS cells (stimulated or unstimulated) adherent to the zirconium alloy generated lower amounts of Factor Xa compared to those bound to the other alloys and unstimulated cells grown on TCPS. These results indicate that ROS cells cultured on these synthetic surfaces differentially express procoagulant activity and that cells grown on TCPS, but not the metallic alloys, exhibit increased TF activity in response to stimulation by calcium ionophore. This procoagulant activity may potentiate subsequent inflammatory responses associated with the use of orthopedic biomaterials and thereby influence the tissue compatibility of the implant. PMID- 9860172 TI - Reactions of cells to topography. AB - Though contact guidance has been known since the very early days of cell culture very little quantitative examination of the reaction of cells to topography has been made. Exceptions to this subjective approach are given prominence below. Yet if we are to understand how cells react and if we are to be able to design ideal substrata for particular cells we need this information. Precision and quantitation are required both of the methods of examination of the cells but also in the definition of that topography. Recently it has become clear that the these reactions occur at the nanometric scale and have importance for use in cellular engineering and tissue repair. Topography appears to provide a set of very powerful signals for cells. PMID- 9860173 TI - Two-dimensional manipulation of confluently cultured vascular endothelial cells using temperature-responsive poly(N-isopropylacrylamide)-grafted surfaces. AB - Temperature-responsive hydration/dehydration changes in surface-grafted poly(N isopropylacrylamide) (PIPAAm) were utilized for hydrophilic/hydrophobic surface property alterations in cell culture. In this report, we utilized PIPAAm-grafted surfaces to recover confluently-cultured vascular endothelial cells as coherent monolayers from this cell culture substrate and to transfer to new cell culture substrates. For this purpose, we used two different methods to recover and transfer cell monolayer cultures: (1) chitin membranes used as an apical side cell support during cultured cell transfer, allowing cell basal side reattachment to new culture substrates after transfer; and (2) a cell culture insert (porous PET) used as both a support as well as new substrate, allowing basal surfaces of cultured cells to be exposed to the medium after transfer. In both cases, all cells grown on PIPAAm-grafted surfaces detach completely with maintenance of basement membrane-like structure. Recovered cells attach to the second culture surfaces, covering more than 60% of the new substrate, and retain approximately 90% viability and their original function as judged from tissue-type plasminogen activator secretion. This technique could be utilized to prepare novel bioartificial organs as well as cell co-culture systems by multi-layering different cell types to mimic tissue structures for tissue engineering. PMID- 9860174 TI - Vascular cell attachment and procoagulant activity on metal alloys. AB - The attachment and growth of vascular smooth muscle cells on biomaterials used as components of devices implanted in the vascular space may influence the biocompatibility of such materials. The nature of the materials may affect the attachment and/or the activation of these cells' procoagulant responses. Therefore, the main objective of this study was to measure the strength of adhesion of these vascular cells to potential biomaterials (titanium, zirconium alloys, and stainless steel) by exposing them to a range of shear stresses (50 300 dyn cm(-2)) in a parallel plate flow chamber. The procoagulant responses of the cells were evaluated by measuring the tissue factor (TF) activity promoted by the different materials under flow conditions. The materials supported distinctly different levels of initial cell adhesion in static culture. However, the fraction of adherent cells did not decline significantly with incrementally increasing shear stress within the range tested. TF expression, as measured by factor Xa (FXa) production. was material-dependent. For example, cells cultured on Ti1313 exhibited more FXa production (13.2 nM 10(-5) cells) than Ti1313(DH) (8.5 nM 10(-5) cells) or stainless steel (2 nM 10(-5) cells). Thus, our studies indicate that the level of adhesion, strength of attachment and the expression of procoagulant activity of adherent vascular cells depend strongly on the nature of the underlying biomaterial. PMID- 9860175 TI - Evaluating the interaction of bacteria with biomaterials using atomic force microscopy. AB - Bacterial infection of biomaterials represents one of the most important reasons for the failure of transdermal or implanted medical devices. The first and least understood step in biomaterial-associated infections is the initial interaction between bacteria and a surface. This initial interaction can be either attractive or repulsive depending on the physiochemical nature of the biological and synthetic surfaces, as well as the properties of the interstitial fluid. We have shown that atomic force microscopy (AFM) can be employed as an exquisitely sensitive and versatile tool for quantifying the interaction between bacteria and surfaces in physiological solutions. The forces of interaction between an AFM cantilever tip and a uniform lawn of bacteria immobilized on glass were determined. By comparing the interactions of cantilever tips with lawns of isogenic E. coli strains carrying genetic lesions that alter their cell surface composition, it was possible to evaluate the effect of macromolecules such as lipopolysaccharide and capsular polysaccharide on the adhesion process. Mutations that result in the synthesis of truncated lipopolysaccharide or in the overproduction of the negatively charged capsular polysaccharide colanic acid render the interaction of the bacteria with the AFM tip unfavorable due to increased electrostatic repulsion. Furthermore, AFM could be used to evaluate the adhesion of bacteria onto commercially relevant biomaterials. In one approach, micron-size polystyrene beads were attached to AFM tips which were then used to measure forces. Unfortunately, this approach is limited by the meager number of materials manufactured as beads of a size suitable for AFM measurements. As an alternative approach, AFM cantilever tips were coated with a confluent layer of bacteria and used to probe planar surfaces. In this configuration, AFM could be employed to measure the force of interaction between virtually any bacterium and surface of interest. PMID- 9860176 TI - Improving endothelial cell adhesion to vascular graft surfaces: clinical need and strategies. AB - Synthetic vascular grafts do not spontaneously endothelialize in humans and require some form of anticoagulation to maintain patency. Preseeding synthetic graft materials such as expanded polytetrafluoroethylene (ePTFE) and polyethylene terephthalate (PET) with endothelial cells (EC) has been examined in various in vitro and in vivo models. Although various studies provide encouraging results, clinical trials for EC seeding on synthetic grafts have not been equally successful. This paper provides a brief review of the various reports on EC seeding in animal and clinical studies. We discuss the inefficiencies associated with the EC seeding process and examine plasma protein treatment of the graft surfaces as a viable option for improving EC attachment, retention and spreading. As an alternative to existing therapies we present data on a heterogeneous ligand treatment of fibronectin (Fn) and avidin-biotin for enhanced human umbilical vein endothelial cell (HUVEC) adhesion to ePTFE graft surfaces. Control consisted of HUVECs seeded on Fn treated ePTFE graft surfaces. Functionality of HUVECs was assessed by measuring prostacyclin production of cells on both homogeneous and heterogeneous ligand treated surfaces. Laminar flow studies with a variable width flow chamber and scanning electron microscopy were used to measure initial cell retention and observe initial cell spreading on ePTFE surfaces, respectively. HUVEC retention on heterogeneous ligand treated graft surface was significantly (p < 0.001) higher compared to homogeneous ligand treated surfaces for shear stress in the range of 10-30 dyn cm(-2). HUVEC showed more cellular spreading on the heterogeneous ligand treated surface after seeding for 1-2 h. In vivo experimentation was performed in immune deficient (nude) rats by replacing a section of both the femoral arteries with 8 mnm long, 1 mm internal diameter denucleated ePTFE grafts treated with homogeneous and heterogeneous ligands respectively. Both grafts were seeded with similar cell density for 15 min prior to implantation. EC attachment and retention was measured by staining EC with hematoxylin and counting the cells before and after flow using light microscopy. The results indicate that a heterogeneous ligand treatment of graft surfaces using avidin-biotin and Fn-integrin attachment mechanisms increase cell seeding efficiency, initial cell retention and cellular spreading. PMID- 9860177 TI - Probing heterotypic cell interactions: hepatocyte function in microfabricated co cultures. AB - Replacement of liver function using extracorporeal bioartificial systems has been attempted with limited success. The instability of the hepatocyte phenotype in vitro has restricted the useful lifetime of these devices. Co-cultivation of hepatocytes with mesenchymal cells is one method that has been widely utilized to stabilize the liver-specific function of isolated cells; however, co-culture has yet to be successfully incorporated in a bioreactor setting. In this study, we probed heterotypic cell interactions in co-cultures of hepatocytes and 3T3 in order to better understand the cellular microenvironment necessary to induce and stabilize liver-specific functions. Using microfabrication and conventional techniques to control the heterotypic interface, the effects of varying degrees of heterotypic interaction on tissue function (albumin and urea synthesis) were examined. Our data indicated maximal induction of liver-specific functions in cultures with maximal initial heterotypic interaction, and that induction of hepatic functions in hepatocytes was increased in the vicinity of fibroblasts as compared to hepatocytes far from the heterotypic interface. Furthermore, our data suggested that heterotypic cell contact is necessary for induction of these functions. These studies will aid in the formation of design criteria for a co culture based bioartificial liver, as well as provide a useful tool to study the role of heterotypic and homotypic interactions in liver physiology and pathophysiology. PMID- 9860178 TI - Cell type-specific modulation of fibronectin adhesion functions on chemically derivatized self-assembled monolayers. AB - Cell type-specific responses (microfilament stress fibers for fibroblasts or neurites for neuroblastoma cells) were evaluated in culture on inert and chemically-derivatized silane substrata adsorbed with fibronectin (Fn). Substrata of self-assembled monolayers contain a 14-17 carbon aliphatic chain terminating with different chemical endgroups -- [CH3], [C=C], [Br], [CN], [Diol], [COOH], [NH2], [SH], [SCOCH3], or [SO3H]. Fn adsorbed effectively to all derivatized surfaces. 3T3 fibroblasts or neuroblastoma cells attached equivalently to all surfaces preadsorbed with Fn, indicating availability of receptor binding sites on Fns. However, transmembrane signaling from Fn(adsorbed): receptor(cell) surface complexes yielded a range of abilities for generating F-actin stress fibers in fibroblasts or neurites in neuroblastoma cells. Efficiency for stress fiber formation was very different from that of neurite extension. The same chemical endgroups on glass, titanium, or germanium yielded the same patterns of cellular physiological responses, indicating that inert substrata do not act at a distance and that only chemical endgroups regulate Fn signaling functions. When adhesion-inert albumin is co-adsorbed with Fn, efficiency of neurite extension is improved on some surfaces or diminished on others. These results indicate that the conformation of Fn(adsorbed) changes in specific ways on derivatized substrata. Change in Fn conformation was confirmed by FTIR/ATR spectroscopy experiments of Fn(adsorbed). Overall, these studies indicate changes in Fn conformations on chemically-derivatized self-assembled monolayers leading to up- or down-regulation of cell type-specific physiological responses from receptors via their signaling pathways. They also offer predictability for regulating responses of specific cell types when these cells interact with biomaterial implants in vivo. PMID- 9860179 TI - Treatment of bacterial biofilms on polymeric biomaterials using antibiotics and ultrasound. AB - New studies on the effect of frequency and duration of exposure upon the ultrasonic enhanced action of gentamicin against biofilms of Escherichia coli are presented in this paper. Ultrasound was applied at clinical levels which would not harm human tissue. The study on frequency indicated that lower frequencies produce higher levels of killing. The results of the timed experiment indicate that complete sterilization of a 14-h biofilm can be achieved after 6 h of exposure. These findings are significant because they show that biofilms can be reduced to zero reproductive ability as assessed by plate counting. PMID- 9860180 TI - Retinal pigment epithelium cell culture on thin biodegradable poly(DL-lactic-co glycolic acid) films. AB - Thin films of 50:50 and 75:25 poly(DL-lactic-co-glycolic acid) (PLGA) were manufactured with a controlled thickness of less than 10 microm. The effect of PLGA copolymer ratio on in vitro cell attachment, proliferation, morphology, and tight junction formation was evaluated using a human D407 retinal pigment epithelium (RPE) cell line. Almost complete cell attachment was achieved on both PLGA films after 8 h of cell seeding, which was comparable to that on tissue culture polystyrene (TCPS) controls. The initial cell seeding density affected attachment, and the optimal value for 50:50 PLGA was 25000 cells cm(-2). After 7 days of in vitro culture, cell density on 50:50 and 75:25 PLGA films increased 45 and 40 folds, respectively, and a 34-fold increase was observed on TCPS. The RPE cells cultured on PLGA films at confluence had a characteristic cobblestone morphology. Confluent RPE cells also developed normal tight junctions in vitro which were concentrated mainly at the apical surfaces of cell-cell junctions. These results demonstrated that thin biodegradable PLGA films can provide suitable substrates for human RPE cell culture, and may serve as temporary carriers for subretinal implantation of organized sheets of RPE. PMID- 9860182 TI - Albumin-binding surfaces: in vitro activity. AB - Immobilized monoclonal antibodies (Mabs) have been used to attract specific molecules to a solid surface from complex mixtures such as blood, plasma or serum, thereby directing the response to the modified substrate, a key goal in rational biomaterial design. The nature of the Mab dictated the nature of the response: anti-albumin antibodies were used to prevent cell and platelet adhesion in vitro, whilst anti-fibronectin Mabs promoted attachment. Patterned surfaces could be formed, bearing Mabs that generated adhesive and non-adhesive regions. Fibrinogen adsorption from plasma showed a Vroman peak on unmodified control polymer, which was reduced by 64% in the presence of surface-bound anti-albumin Mab. Immobilization of a control Mab reduced fibrinogen adsorption only slightly, implying an albumin-mediated effect. In static tests, platelet adhesion from human platelet rich plasma was significantly reduced by the immobilization of anti-HSA Mab when compared to the untreated FEP surface (p < 0.0001). This effect was also seen with citrated blood flowing through Mab-treated polyurethane tubing at a shear rate of 132 s(-1) (p=0.034). Since platelets and proteins (as blood, plasma or serum) were introduced to the surface simultaneously, the generation of a defined protein film must have been sufficiently rapid as to shape the platelet or cell response. PMID- 9860181 TI - Albumin-binding surfaces: synthesis and characterization. AB - The nature of the proteinaceous film deposited on a biomaterial surface following implantation is a key determinant of the subsequent biological response. To achieve selectivity in the formation of this film, monoclonal antibodies have been coupled to a range of solid substrates using avidin-biotin technology. Antibody clones varied in their antigen-binding activity following insertion of biotin groups into lysine residues. Biotinylated antibodies coupled to solid substrates via an immobilized avidin bridge retained their biological activity. During immobilization of avidin a significant proportion of the protein molecules were passively adsorbed rather than covalently attached to the surface. This loosely bound material could be removed by stringent elution procedures which resulted in a surface density of 5.4 pmol avidin cm(-2). Although these conditions would be harsh enough to denature monoclonal antibodies, they did not destroy the biotin-binding activity of the residual surface-coupled avidin, enabling the subsequent immobilization of biotinylated antibodies. The two-step immobilization technique allowed the use of gentle protein modification procedures, reduced the risk of surface-induced denaturation and removed loosely bound material from the surface. The versatility of the technique encourages its application to a wide range of immobilization systems where retention of biological activity is a key requirement. PMID- 9860183 TI - Retinal pigmented epithelium cultures on thermally responsive polymer porous substrates. AB - A cross-linkable co-polymer of UV-sensitive 4-(N-cinnamoylcarbamide)methylstyrene (CCMS) and N-isopropylacrylamide (NIPAAm), was applied to porous tissue culture inserts. Surface chemical analyses of the inserts show an introduction of a thermally responsive polymer comparable to that on similarly incorporated non porous polystyrene surfaces. Contact angle measurements as well as atomic force microscopy show a surface change in response to changing temperature in an aqueous environment, from hydrophilic, extended polymer chains below 32 degrees C to a dense hydrophobic film above 32 degrees C. Cell growth on porous inserts allowed measurement of cell expression, such as transepithelial resistance and fluid transport, which are not observable on cells from non-porous surfaces. Cultures of retinal pigmented epithelium (RPE) were able to restore an environment similar to in vivo by forming a tight junction barrier membrane upon confluence at 37 degrees C, as observed by changes in morphology, transepithelial resistance, and directionally-specific fluid transport. In addition, cells cultured on these surfaces detached as an oriented polarized sheet when the inserts were brought to 20 degrees C. This cell sheet was transplanted to other tissue culture surface without polymer detachment or dissolution, or cell damage caused by traditional detachment methods using proteolytic enzymes. PMID- 9860184 TI - Production and characterisation of monoclonal antibodies specific for bovine interleukin-4. AB - Genetic immunisation is a simple method for producing polyclonal antibodies in mice. By this method, we produced antibodies against bovine interleukin-4 (BoIL 4). After a final injection with a recombinant BoIL-4 protein, nine stable hybridoma cell lines were established which secreted monoclonal antibodies (MAbs) against this cytokine. Specific binding of each of the MAbs to recombinant BoIL-4 produced by Escherichia coli, baculovirus, and Trypanosoma brucei was demonstrated in an indirect ELISA and/or in Western blotting. These MAbs recognise the same antigenic region localised in the first 47 amino acids of the mature protein. None of them was able to neutralise the biological activity of the BoIL-4 under the conditions tested but one allowed the detection of BoIL-4 by flow cytometry. PMID- 9860185 TI - Effect of vaccination route and composition of DNA vaccine on the induction of protective immunity against pseudorabies infection in pigs. AB - Vaccination with naked DNA may be an alternative to conventional vaccines because it combines the efficacy of attenuated vaccines with the biological safety of inactivated vaccines. We recently showed that the vaccination with naked DNA coding for the immunorelevant glycoprotein D (gD) of pseudorabies virus (PRV) induced both antibody and cell-mediated immunity in pigs and provided protection against challenge infection. To determine whether the efficacy of the naked DNA vaccination against PRV could be improved, we compared three sets of variables. First, the efficacy of the naked DNA vaccine coding only for the immunorelevant gD was compared with a cocktail vaccine containing additional plasmids coding for two other immunorelevant glycoproteins, gB and gC. Second, the intramuscular route of vaccination was compared with the intradermal route. Third, the commonly used needle method of inoculation was compared with the needleless Pigjet injector method. Five groups of five pigs were vaccinated three times at 4-weeks intervals and challenged with the virulent NIA-3 strain of PRV 6 weeks after the last vaccination. Results showed that although the cocktail vaccine induced stronger cell-mediated immune responses than the vaccine containing only gD plasmid, both vaccines protected pigs equally well against challenge infection. Intradermal inoculation with a needle induced significantly stronger antibody and cell-mediated immune responses and better protection against challenge infection than intramuscular inoculation. Our data show that the route of administering DNA vaccines in pigs is important for an optimal induction of protective immunity. PMID- 9860186 TI - Interferon induction in turkeys by oral administration of the imidazoquinolinamine S-28828 and modulation of the pathogenesis of Escherichia coli. AB - A synthetic imidazoquinolinamine, S-28828, has been shown to be an effective antiviral and antitumor agent in mammals. This immune modifier induces a number of cytokines such as interferons, tumor necrosis factor-alpha, interleukins and granulocyte-macrophage colony-stimulating factors in mammals. We showed that when turkeys were given S-28828 orally, high serum titers of IFN were induced in a dose-dependent manner. Turkeys, once stimulated by S-28828, became refractory to IFN production by repeated stimulation. S-28828 induced spleen, bone marrow and peripheral leukocytes to produce IFN in vitro. Splenic adherent cells were the main producers of IFN after in vitro stimulation. S-28828-induced IFN was identified as type I IFN that was pH-resistant but heat-labile. We examined the effect of a high dose (100 mg kg(-1) body weight) of S-28828 on the pathogenesis of E. coli in turkeys. Treatment with S-28828 increased mortality in infected birds and impaired E. coli clearance from the liver. The enhancement of the pathogenicity of E. coli by S-28828 may have been due to the massive release of cytokines inducing a shock-like syndrome in infected turkeys. PMID- 9860187 TI - Genes of chicken MHC regulate the adherence activity of blood monocytes in Rous sarcomas progressing and regressing lines. AB - The influence of the chicken major histocompatibility (B) complex (MHC) on the adherence potential of monocyte-derived macrophages was examined using the congenic chicken lines CB and CC. These lines represent well-defined genetic models for the study of resistance (CB) or susceptibility (CC) to the progressive growth of Rous sarcomas. Using a monoclonal antibody specific for chicken monocytes/macrophages, CB and CC chickens were shown by flow cytometry analyses to have similar proportions of peripheral blood monocytes. However, when the glass-adherence potential of these cells was compared during incubation in tissue culture medium over 24, 48 and 72 h at 40 degrees C, significant differences were seen between cells from these two inbred lines. After 24 and 48 h, glass adherence by CB cells was 2-3 fold higher than that of CC cells. After 72 h this difference decreased to 1.5 fold. At 24 and 48 h, the adherent CB macrophages also appeared about 1.5 times larger than those of CC chickens. Genetic analysis using F1 hybrids (CBxCC) showed that this trait is regulated by a dominant gene that segregates with the B12 haplotype in the backcross generation F1xCC. From the results obtained with the recombinant congenic lines CB.R1 and CC.R1, we conclude that the gene regulating adherence potential is localized within the B F/L region of the chicken MHC. About 50% of adherent cells were able to phagocytose opsonised FITC-labelled Zymosan particles. The level of nitric oxide production in vitro by CB and CC macrophages was equal. The importance of cells of the mononuclear phagocyte system for the response to Rous sarcoma virus (RSV) infection was studied in CB chickens using the anti-macrophage agents silica, carrageenan, and C12MDP, encapsulated in liposomes. In those chickens treated with silica and carrageenan, we observed progressive growth of RSV-induced tumors. The graft-versus-host reactivity of peripheral blood lymphocytes (PBL) of treated chickens was comparable to controls. In vitro nitric oxide production by macrophages from silica-treated chickens was higher than by macrophages from untreated controls. PMID- 9860188 TI - Effect of various adjuvants on secondary immune response in chickens. AB - Stimulatory effects of several types of adjuvants on secondary antibody response to inactivated Newcastle disease virus (iNDV) were examined in chickens. For this purpose, animals were primed with iNDV without adjuvant resulting in a low but significant antibody response, boosted with iNDV plus adjuvant 3 weeks later, and analysed for specific antibody titres in serum 3 weeks after the booster. Water in-mineral oil emulsion (W/O) caused significant increase in antibody titres measured in an indirect enzyme-linked immunosorbent (ELISA), haemagglutination inhibition (HI), and virus neutralisation (VN) assay. The adjuvants tested included three oil-in-water emulsions (i.e. mineral oil-in-water, sulpholipo(SL) Ficoll400/squalane-in-water and sulpholipo-cyclodextrin/squalane-in-water), three negatively-charged polymers with high molecular weight (i.e. polyacrylate, polystyrenesulphonate and sulpho(S)-Ficoll400) and two surface-active agents (i.e. dimethyldioctadecylammonium bromide (DDA) and Quil A). These adjuvants enhanced significantly the secondary immune response but none reached the titre obtained with W/O. Combinations of adjuvants with distinct physicochemical properties, i.e. polyacrylate and DDA revealed only slight, beneficial effects. We concluded that the various types of adjuvants tested can stimulate secondary immune responses in primed animals but that W/O is superior. PMID- 9860189 TI - T lymphocytes, B lymphocytes, and macrophages in the ovaries and oviducts of laying hens experimentally infected with Salmonella enteritidis. AB - Subsets of T lymphocytes, B lymphocytes, and macrophages in the ovaries and oviducts of laying hens were enumerated by immunohistochemistry after intravenous inoculation with Salmonella enteritidis. Almost all T cell subsets in the ovaries and different regions of the oviduct increased in number at 7 days post inoculation and reached a peak by day 10. This T cell surge was followed by a peak in B cell numbers at day 14. The number of macrophages declined initially but recovered to preinoculation levels by day 21. At day 21, the numbers of T and B cells also returned to normal levels, except for IgG+ B cells in the infundibulum, isthmus, and vagina where they remained consistently elevated. The T and B cell proliferation at 10-14 days post-inoculation immediately preceded a decline in the number of S. enteritidis positive tissues from infected hens beginning at day 14 suggesting that these lymphocytes play a major role in the local immune response to S. enteritidis. The Salmonella-oviduct model will be useful for future studies on local immunity to various infectious agents. PMID- 9860190 TI - Effects of in vitro addition of exogenous vitamins C and E on gilthead seabream (Sparus aurata L.) phagocytes. AB - The phagocytic cell function is an important factor of resistance to infection in fish. Although it is known that several nutritional factors affect phagocyte function and that vitamins C and E modulate the vertebrate immune system, there are no data on the direct effect of these vitamins or on their possible synergistic effects on the fish phagocyte functions. We report the in vitro effect of vitamin C and/or E on the functions and ultrastructure of gilthead seabream (Sparus aurata L.) head-kidney phagocytic leukocytes. Head-kidney leukocytes were incubated with different concentrations of vitamins C and/or E for 48 h and then their migration, respiratory burst, phagocytosis and ultrastructure were studied. Leukocyte migration and phagocytosis increased when vitamin C or E were added, while the respiratory burst was synergistically enhanced when both vitamins were used together. Noticeable ultrastructural changes not previously reported were induced by these vitamins in gilthead seabream phagocytes (acidophilic granulocytes and macrophages). These results indicated that vitamins C and E added in vitro modulate the main functions of gilthead seabream phagocytes and induced ultrastructural changes characteristic of cell activation on such cells. PMID- 9860192 TI - Ventricular repolarization time indexes following anthracycline treatment. AB - The anthracyclines, doxorubicin and daunorubicin, are antibiotics effective in the treatment of many malignancies. However, their usefulness is limited by the development of potentially fatal cardiotoxicity. Cardiac monitoring by a noninvasive test capable of identifying patients at high risk of cardiac damage, before the ejection fraction deteriorates would have clinical utility. Electrocardiograms and echocardiograms are routinely utilized for noninvasive assessment of myocardial function. However, of the ECG abnormalities described, none has been noted to be of consistent predictive value for cardiotoxicity. The aim of this study was to assess the effects of doxorubicin on ventricular repolarization time indexes, as they have been shown to be effective in the identification of electrical myocardial instability and, hence, in the identification of risk for either arrhythmia or heart failure. For this reason, electrocardiograms were compared in 35 cancer patients at the first presentation (drug-free state) and after 29.4 +/- 37.65 weeks of treatment with doxorubicin. The results of the present study showed that after only a short period of treatment with doxorubicin there was a significant increase in ventricular recovery time dispersion indexes (QTc, JT, and JTc dispersion, and their "adjusted" values). Thus, increased regional variation in ventricular repolarization could be, in the absence of a significant modification of the echocardiographic parameters, an early marker of an electropathy, due to the early cardiotoxic action of doxorubicin on myocardial cells, eventually leading to heart failure. PMID- 9860191 TI - Functional activity of the CFTR Cl- channel in human myocardium. AB - The cyclic AMP (cAMP)-dependent chloride channel in the heart has been identified in various species as the cystic fibrosis transmembrane conductance regulator (CFTR). Although functional expression of the channel in the human atrium has been reported, we could not induce any cAMP-dependent chloride conductance in the atrial cells even with maximal cAMP stimulation, whereas the conductance could be induced in rabbit ventricular cells. To clarify the discrepancy between the results, we examined the level of CFTR mRNA expression in both conductance positive (human colonic epithelium and rabbit ventricle) and -negative (human atrium) tissues. Total RNA samples prepared from these tissues were subjected to the reverse transcription-polymerase chain reaction (RT-PCR). While CFTR transcripts were amplified from the conductance-positive samples, no amplified products could be detected from the conductance-negative sample. A nested PCR performed on the RT-PCR products of the conductance-negative sample resulted in successful amplification of the transcripts, indicating that the level of the CFTR mRNA expression in human atrium is extremely low compared with that in colonic epithelium and rabbit ventricle. The same molecular results were observed in human ventricular tissues. A nucleotide sequencing of the amplified transcripts showed that exon 5 of the CFTR gene was not alternatively spliced in human atrium and ventricle, and both the exon 5 spliced and unspliced isoforms were expressed in rabbit ventricle, unlike the findings of previous reports. Our data suggest that the amount of CFTR expressed in human myocardium might be physiologically insufficient to activate detectable cAMP-dependent chloride conductance. PMID- 9860193 TI - A comparison of ultrastructural changes on endomyocardial biopsy specimens obtained from patients with diabetes mellitus with and without hypertension. AB - The pathogenesis of diabetic cardiomyopathy is unknown. The synergistic, or enhanced, effect of hypertension on pathological changes in the heart of diabetic patients has been highly suspected. The purpose of this study was to evaluate the myocardial changes related to diabetes mellitus with and without hypertension, using biopsy specimens. We examined the ultrastructural changes in biopsy specimens of the endomyocardium obtained from 25 patients. They were divided into four groups: controls without hypertension or diabetes mellitus (n = 6), and patient with hypertension (n = 3), diabetes mellitus (n = 8), and diabetes with hypertension (n = 8). The diabetic patients showed nearly normal or mildly depressed systolic left ventricular function. Ultrastructural pictures were analyzed for thickening of the capillary basement membrane, presence of toluidine blue-positive materials (i.e., materials showing metachromasia) in the myocytes, size of myocytes, and interstitial fibrosis. The thickening of the capillary basement membrane, the accumulation of toluidine blue-positive materials, and interstitial fibrosis were all significantly greater in the patients with diabetes mellitus compared to the control subjects. The myocytes tended to be small (cell atrophy) in the diabetes group. Although these pathological changes in the heart were characteristic of diabetic patients, irrespective of the presence or absence of hypertension, the presence of hypertension increased the pathological changes of myocardial cells as well as abnormality in the capillary vessels in patients with diabetes mellitus. Alterations in the myocardial cells and capillaries, caused by diabetes mellitus, may lead to myocardial cell injury and interstitial fibrosis and, ultimately, to ventricular systolic and diastolic dysfunction, especially when the diabetes is accompanied by hypertension. PMID- 9860194 TI - Expression of cell division cycle 2 kinase transcription in chronically rejected cardiac allografts of nonhuman primates. AB - Accurate diagnosis and prevention of graft arteriosclerosis, known as chronic rejection, is critical to the success of cardiac transplantation, but often is difficult to attain. Expression of cell division cycle (cdc) 2 kinase, which plays a critical role in cell transition through the G2/M phase, is critical to the proliferation of vascular smooth muscle cells. To evaluate the usefulness of cdc2 kinase expression for pathophysiological analysis of chronic rejection, heterotopic cardiac transplantation was performed in Japanese monkeys (n = 7). Standard reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ RT PCR were performed to evaluate the expression. In the coronary arteries of chronically rejected allografts, enhanced cdc2 kinase expression was observed in thickened intima and media, while none was expressed in native hearts. The cdc2 kinase was also expressed before the intimal thickening occurred. These results indicate that enhanced expression of cdc2 kinase is a sensitive indicator for chronic cardiac rejection; targeting cdc2 kinase may be a viable gene therapy for prevention of this vasculopathy. PMID- 9860195 TI - Calcium equally increases the internal calcium recirculation fraction before and after beta-blockade in canine left ventricles. AB - We studied whether intracoronary Ca administration after beta-blockade would increase the internal Ca recirculation fraction (RF) analogously to the Ca administration before beta-blockade. This was performed in excised cross circulated canine hearts. We analyzed the exponential decay component of the postextrasystolic potentiation (PESP) following a spontaneous extrasystole. All the PESPs decayed in alternans with atrial pacing at a constant rate. We obtained the time constant (tau(e)) of the monoexponential decay component of the alternans PESP. An increment of intracoronary Ca by 1.5 mmol/l enhanced the left ventricular contractility index Emax (end-systolic maximum elastance) by 2.5 times before and after beta-blockade with propranolol. The intracoronary Ca after beta-blockade slightly but significantly increased tau(e), and hence increased RF calculated from tau(e) by RF = exp(-1/tau(e)). This was analogous to the slightly increased tau(e) and RF with Ca before beta-blockade. We speculate that the myocardial cyclic AMP-dependent phosphorylation level would not significantly alter the effect of intracoronarily administered Ca on myocardial Ca handling, in terms of tau(e) and RF. PMID- 9860196 TI - Role of K+ channels in EDHF-dependent relaxation induced by acetylcholine in canine coronary artery. AB - To identify the K+ channels responsible for endothelium-derived hyperpolarizing factor (EDHF)-dependent relaxation, we studied the effects of various K+ channel blockers on acetylcholine-induced relaxation, which persists even in the presence of both an inhibitor of nitric oxide synthase and that of cyclooxygenase, in canine coronary artery rings. A nonselective K+ channel blocker, tetrabutylammonium (TBA), a large and intermediate conductance Ca2+-activated K+ channel blocker, charybdotoxin (CTX), and a voltage-dependent K+ channel blocker, 4-aminopyridine (4-AP), significantly inhibited this residual relaxation. A combined treatment with CTX and 4-AP almost completely blocked the relaxation. Neither a large (iberiotoxin) nor a small (apamin) conductance Ca2+-activated K+ channel blocker blocked the relaxation. We also investigated effects of K+ channel blockers on basal tone to determine whether or not EDHF is involved in regulating basal tone. TBA and CTX substantially raised basal tone to a greater degree in endothelium-intact preparations than in endothelium-denuded preparations. These results indicate that EDHF may exert its relaxing action through intermediate conductance Ca2+-activated and voltage-dependent K+ channels in canine coronary arteries. In addition, EDHF may play a role in maintaining basal vascular tone. PMID- 9860197 TI - Modulation of cardiac interstitial noradrenaline levels through K(ATP) channels during ischemic preconditioning in rabbits: comparison of the effect of anesthesia between pentobarbital and ketamine + xylazine. AB - In rabbits, both the stimulation of alpha1-adrenoceptors and ischemic preconditioning (PC) reduce infarct size. One candidate for the mechanism of PC is noradrenaline (NA), which stimulates alpha1-adrenoceptors in the myocardium during PC. Opening of the K(ATP) channel is considered to be another candidate for PC, since a K(ATP) channel blocker, glibenclamide, blocks the infarct size reducing effect of the PC of 5-min ischemia and 5-min reperfusion in rabbits anesthetized with ketamine + xylazine. However, in rabbits anesthetized with pentobarbital, the infarct size-reducing effect of PC was not blocked by glibenclamide. The effect of glibenclamide on the PC effect thus differs depending on the anesthesia used. Therefore, we speculated that the increase in cardiac interstitial NA levels induced by PC may be modified by the anesthesia used, thus regulating the effect of glibenclamide on the PC effect. In open-chest Japanese white male rabbits anesthetized with pentobarbital or ketamine + xylazine, myocardial interstitial NA levels were measured before and during the PC of 5-min ischemia and 5-min reperfusion in the presence or absence of the K(ATP) channel blocker, glibenclamide (0.3 mg/kg, i.v.), using a microdialysis technique. The NA levels were measured using high-performance liquid chromatography coupled with electrochemical detection. The PC of 5-min ischemia and 5-min reperfusion significantly elevated the interstitial NA level. This increase in the NA level was not blocked by glibenclamide under anesthesia with pentobarbital. Under anesthesia with ketamine + xylazine, the PC did not cause an increase in the myocardial interstitial NA level in either the absence or the presence of glibenclamide. In conclusion, PC elevates the myocardial interstitial NA level, and this elevation is not mediated through the opening of the K(ATP) channel under anesthesia with pentobarbital. Under anesthesia with ketamine + xylazine, PC does not cause an increase in the myocardial interstitial NA level. This may explain the discrepancy in the blocking effect of glibenclamide on the infarct size-reducing effect of PC between anesthesia with pentobarbital and ketamine + xylazine. PMID- 9860198 TI - Vascular effects of endothelin-1 in stage 21 chick embryos. AB - Endothelin-1 is a very potent vasoconstrictor, but its function has not yet been investigated in the early stage of cardiovascular development. The purpose of the present study was to clarify whether endothelin-1 exerts a hemodynamic effect in stage 21 chick embryos. We measured vitelline artery blood pressure with a servo null micropressure system and blood flow velocity at the dorsal aorta with a 20 MHz pulsed Doppler velocity meter. The vitelline vessels were directly measured with a microscope video system. While monitoring these parameters, endothelin-1 was infused into a vein by a microinjector and data were collected. Endothelin-1 increased the blood pressure and heart rate, but decreased the dorsal aortic blood flow. Only the vitelline veins with a diameter of between 100 and 200 microm constricted after endothelin infusion, but smaller or larger veins and the arteries did not show any significant change in size, although the resistant arteries could not be measured by this method. In conclusion, endothelin-1 has apparent constrictive effects in the selected vessel in the early stages of cardiovascular development when the endocrine and autonomic nervous systems have not yet developed. PMID- 9860199 TI - Acute ascending aortic dissection complicating open heart surgery: cerebral perfusion defines the outcome. AB - OBJECTIVE: This retrospective study was designed to assess the risks of acute ascending aorta dissection (AAD) as a rare but potentially fatal complication of open heart surgery. METHOD: Among 8624 cardiac surgical procedures under cardiopulmonary bypass (CPB) and cardioplegic myocardial protection from 1978 to 1997, 10 patients (0.12%) presented with a secondary or so called 'iatrogenic' AAD. There were seven men and three women, mean age 64 +/- 9 years, ranging from 47 to 79. The original procedures involved five coronary artery bypass grafts (CABG), one repeat CABG, one aortic valve replacement (AVR), one AVR and CABG, one mitral valvuloplasty (MVP) and CABG and one ascending aorta replacement. We retrospectively analyzed their hospital records. RESULTS: Group I consisted of seven patients with AAD intraoperatively and group II consisted of three patients who developed acute AAD 8-32 days after cardiac surgery. In group I, treatment consisted of the original procedure, plus grafting of the ascending aorta in six patients and closed plication and aortic wrapping in one. In group II, two patients received a dacron graft and one patient developed lethal tamponnade due to aortic rupture before surgery. Postoperatively, six patients responded well and three died (33%), two patients from group I on the 2nd postoperative day with severe post-anoxic encephalopathy, and one from group II with severe peroperative cardiogenic shock. CONCLUSION: Preventing AAD with the appropriate means remains standard practice in cardiac surgery. If AAD occurs, it requires prompt diagnosis and interposition graft to allow a better prognosis. Intraoperative AAD happens at the beginning of CPB jeopardizing perfusion of the supra-aortic arteries. PMID- 9860200 TI - Localized fibrous tumours of the pleura: 15 new cases and review of the literature. AB - OBJECTIVE: To present a series of localized fibrous tumours of the pleura (LFTP), to define the clinical and histopathological diagnostic criteria of this tumour, and to determine the optimal treatment and follow-up. METHODS: Review of the charts of the patients with the diagnosis of LFTP (formerly called benign fibrous mesothelioma), as well as of all the histological sections, including immunohistochemical stains. Review of the literature with special emphasis on the clinical and histological criteria of malignancy. RESULTS: During the last 30 years, we found 15 patients with a complete clinical chart and histological material, particularly paraffin blocks of the tumour. The mean age was 57 years (range 27-79). Eight patients were asymptomatic, and the remaining seven presented with non-specific symptoms. All but one had complete resection of the tumour, including partial lung resection in two and partial chest wall resection in three. The diagnosis was confirmed by histological review in 15 cases. Immunohistochemical stainings showed positivity for vimentin in all cases, for CD 34 in 80%, but were consistently negative for cytokeratins. Nine tumours were histologically classified as malignant. Among them, five recurred, two of which were responsible for death. One benign tumour recurred after 1 year, and was treated successfully by repeat resection and radiotherapy. Overall, 13 patients (86%) were alive with no evidence of disease between 10 months and 27 years after the first resection. CONCLUSIONS: LFTP is a rare tumour which has a benign clinical course in over 80% of the cases, and is asymptomatic in half the patients. The diagnosis is difficult to establish before operation. Treatment consists of complete resection including adjacent structures if necessary. The clinical behaviour of LFTP cannot be predicted on the basis of histological aspects only. If histologically malignant tumours are more prone to recurrence and poor outcome, broad-based and locally invasive tumours bear a higher risk of recurrence. Long term follow-up is therefore mandatory in all cases in order to perform early re-resection when recurrence occurs. PMID- 9860201 TI - Chylothorax following oesophagogastrectomy for malignant disease. AB - OBJECTIVE: To define the incidence, causes, management and impact of Chylothorax after oesophagogastrectomy for malignant disease in Nottingham Thoracic Surgery unit. PATIENTS AND METHODS: Retrospective analysis of 523 patients with cancer of the oesophagus or the gastro-oesophageal junction who underwent oesophageal resection between January 1987 and November 1997 in a single unit using similar techniques and uniform routine perioperative management. RESULTS: Chylothorax occurred in 21 patients (4.0%). There were 12 males and 9 females with a mean age of 64.7 years (SD 7.5). Age, sex, tumour site, length, histological type, depth of wall penetration, nodal status and type of operative approach were not significant predisposing factors on univariate and multivariate analysis. Seventeen patients were treated conservatively (four deaths, 23.5%) and four surgically (one death, 25.0%), effective control of the chylous leak being achieved in all four cases. Eleven patients with a chylous drainage of up to 2.2 l/day, diminishing within 1 week of conservative treatment had an uneventful recovery. However, a chylous drainage of more than 2.5 l/day in the remaining ten patients was associated with increased morbidity, hospital stay, operative mortality and the need for surgical intervention. In comparison with the remaining patients (n = 502), those who developed chylothorax (n = 21) had more respiratory complications (42.8%, P = 0.008), longer mean hospital stay (23.8 days, P = 0.004), higher operative mortality (23.1%, P = 0.004) and, unexpectedly, reduced 5 year survival rate (P < 0.0001). CONCLUSIONS: There appeared to be no clear predisposing factor in the development of a chylous leak other than the routine extensive dissection. Although definitive conclusions can not be drawn, where there is early reduction of the initial amount (in this series up to 2.2 l/day) of drainage, there may be a place for successful non surgical management; in cases of high output chylothorax, persisting after a few days of conservative treatment, however, early re-operation and ligation of the thoracic duct, seems to be advisable. PMID- 9860202 TI - The end of the cold era: from intermittent cold to intermittent warm blood cardioplegia. AB - BACKGROUND: A major reduction in the energy demand of the myocardium results from the electromechanical arrest, and cooling contributes to a lesser degree to this reduction. It is from this assumption that strategies of myocardial protection, utilizing warm blood cardioplegic induction, followed by cold cardioplegia with terminal warm reperfusion before removal of the aortic cross clamp, became established as optimal myocardial protection. Continuous normothermic perfusion 'closed the loop' by avoiding myocardial ischemia and linking warm induction and terminal reperfusion. A series of laboratory and clinical data confirmed the benefits of warm heart surgery on myocardial function and metabolism. The disadvantages of continuous warm blood cardioplegia including disturbance of the operative field, led surgeons to administer warm hyperkalaemic blood intermittently as a new cardioplegic strategy. METHODS: This review examines the laboratory and clinical data with reference to the intermittent warm blood cardioplegia, to establish its experimental basis and place in clinical practice. CONCLUSIONS: Experimental observation and clinical application have established intermittent warm blood cardioplegia as a practical, effective and cheap myocardial protection technique, particularly with reference to coronary artery surgery. PMID- 9860203 TI - Can visual assessment of flow waveform morphology detect anastomotic error in off pump coronary artery bypass grafting? AB - OBJECTIVE: Flow probes are being used for intraoperative assessment of anastomotic quality during off-pump coronary artery bypass grafting (CABG). We conducted a survey with the cooperation of 19 international surgeons to assess the ability of surgeons to detect anastomotic errors by evaluating mean flow and flow waveform morphology. MATERIAL AND METHODS: Mongrel dogs underwent mammary to left anterior descending (LAD) grafting. Mean graft flow and flow wave morphology for varying degrees of anastomotic stenoses were recorded using transit-time flow probes. A questionnaire consisting of ten different recorded flow tracings and the corresponding mean flows were given to 20 surgeons from around the world. The surgeons were asked to determine the degree of stenosis and whether they would re do the anastomosis based upon the mean flow and the flow tracings. RESULTS: All of the 19 surgeons that responded were able to clearly identify a highly stenotic graft (>90% stenosis). However, 24% would re-do a fully patent anastomosis, 58% accepted an anastomosis with moderate stenosis, and 72% accepted anastomoses with severe stenosis. CONCLUSIONS: Evaluation of flow tracing morphology and/or mean flows can be used to reliably detect nearly occluded anastomoses (>90% stenosis). However, surgeons should be cautious in assessing anastomoses with lesser degrees of stenosis, as they may be more difficult to reliably interpret. PMID- 9860204 TI - First cardiological or cardiosurgical reintervention for ischemic heart disease after primary coronary artery bypass grafting. AB - OBJECTIVE: To study the first reintervention for ischemic heart disease anytime after coronary artery bypass grafting (CABG) and the variables that drive its need or bias its occurrence. Reintervention is defined as an isolated or combined repeat surgical or cardiological procedure for ischemic heart disease. METHODS: A consecutive series of 9600 CABG patients (1971-1992) were followed for up to 20 years (99.9% complete). A multivariable time-related analysis was performed. RESULTS: The 1-, 10- and 15-year freedom from reintervention was 99, 89 and 72% respectively. A three-phase hazard function was identified. Patient variables influencing early freedom included anginal instability, completeness of revascularization and institutional variables. Late freedom was influenced importantly by demographic variables, cardiac and non-cardiac comorbidity and extensive arterial grafting. The 1-month and 10-year survival after reintervention was 95 and 73%. The 1- and 10-year freedom from angina after reintervention was 74 and 32%. CONCLUSION: Reinterventions for ischemic heart disease by interventional cardiology or surgery are rather infrequent in the first decade after CABG but nearly half the patients surviving their second decade undergo one. The increased reintervention rate, apparent after 1985 did not go parallel with improved late post-CABG survival. Older age and the presence of multiple arterial grafts seem to reduce but also to bias the event. The very good survival, only when return of angina is present, suggests a more restrictive differential therapy approach, certainly in the presence of a well functioning arterial graft to the antero-septal region and where the co-morbidity might induce a high reinterventional survival cost. PMID- 9860205 TI - Beneficial effects of inhaled nitric oxide in hypoxaemic patients after coronary artery bypass surgery. AB - OBJECTIVE: Arterial oxygenation may be impaired in the early period after open heart surgery, with an associated increase in ventilation time, morbidity and hospital stay. We tested the hypothesis that inhaled nitric oxide could be a useful therapeutic adjunct in this setting. We sought to establish clinical benefits (if any), safety and the appropriate dose range of inhaled nitric oxide therapy in hypoxaemic patients after coronary artery bypass graft surgery. METHODS: Forty patients who satisfied our definition of post-operative impaired oxygenation were prospectively randomised. The treatment group (n = 20) received nitric oxide in addition to ventilatory support. While the control group (n = 20) was managed only by conventional ventilatory support. Cardio-respiratory parameters and clinical outcome measures were compared. RESULTS: We determined the optimum concentration of inhaled nitric oxide as 20 ppm in the majority (60%) of patients. Treatment improved arterial oxygenation (8.4 +/- 1.4 kPa before, 11.8 +/- 1.5 kPa after 4 h, P < 0.001) and this benefit was sustained with lower oxygen fractions required at 24 h (P < 0.001). A significantly shorter period of mechanical ventilation was required in the treatment group (mean ventilation hours 67.0 +/- 5.9 vs. 85.0 +/- 6.5, P < 0.05), although the study did not have the power to distinguish differences in ITU or overall hospital stay. Nitrous oxide and met-haemoglobin levels did not rise appreciably. CONCLUSION: We have established the safety and efficacy of inhaled nitric oxide, at a dose of between 10 and 30 ppm, in this group of patients. We suggest that nitric oxide and a delivery system are useful adjuvants in a cardiac surgical intensive care unit. PMID- 9860206 TI - Inhaled nitric oxide and inhaled prostaglandin E1: effect on left ventricular contractility when used for treatment of experimental pulmonary hypertension. AB - OBJECTIVE: Pulmonary hypertension (PHT) is a life-threatening complication after isolated heart and lung transplantation. Recent work has shown that inhaled nitric oxide (NO) in combination with inhaled prostaglandin E1 (PGE1) reduce pulmonary hypertension but their influence on cardiac contractility is less well defined. METHODS: This study investigated left ventricular contractility as measured by the 'Preload Recruitable Stroke Work-Relation' (PRSW) in 24 anesthetized open chest pigs, 12 receiving in random order NO (50 ppm), PGE1 (20 microg/ml) and their combination compared to 12 controls. PHT was induced by embolization with glass beads (500 microm). Prior to induction of PHT, sonomicrometric crystals were placed on the heart to measure instantaneous cardiac dimensions. Instantaneous intraventricular pressure (micro-tip catheter) and intraventricular dimensions were recorded digitally, while intraventricular volumes were calculated from the intraventricular dimensions applying the cylindric ellipsoidal volume model for the left ventricle. PRSW was calculated from the instantaneous pressure and volume data during rapid vena caval occlusion by analysis of generated pressure-volume loops. All data were analyzed by MANOVA and corrected for heart rate (level of significance #: P < 0.05); PRSW-slope measures contractility, (PRSW-X-intercept did not change significantly). RESULTS: PRSW-change +/- SEM (in percent of initial PRSW after induction of PHT) was 14.6% +/- 4.4% versus 1.6% +/- 4.4% for NO versus Control (P = 0.004), -8.8% +/- 4.6% versus 1% +/- 3.3% (P = 0.18) for PGE1 versus Control and -5.7% +/- 4.4% versus 2.5% +/- 4.2% for NO + PGE1 versus Control (P = 0.33), respectively. In summary, application of NO 50 ppm significantly reduced left ventricular contractility while PGE1 20 microg/ml and the combination of NO and PGE1 did not. CONCLUSION: If NO is not available, the sole application of nebulized PGE1 (20 microg/ml) appears to be safe with respect to left ventricular contractility in the setting of PHT. The combination of NO and PGE1 for the treatment of pulmonary hypertension should be considered for clinical application in situations where a combination of pulmonary hypertension and decreased left ventricular function is present. PMID- 9860207 TI - Endothelial prostacyclin (PGI-2) production of human and porcine valve allografts related to ischemic history. AB - BACKGROUND: The significance of cellular viability in human valve allografts for functional clinical longevity continues to be debated. Meaningful tests for this biological entity are therefore in demand to quantify the relative merits of graft origin and procurement techniques. The valve leaflet endothelium is recognized as a particularly sensitive target to noxes and its continued ability to produce prostacyclin (PGI-2) after explantation has been suggested as indicating viability. OBJECTIVE: Graft ischemic history and species differences were therefore studied in human and porcine valve leaflets by the measurement of endothelial prostacyclin production, post-explantational, basal and after stimulation with bradykinin. METHODS: Four groups of aortic valve donors were established. Fresh human heart-beating donors (h-HBD), cadaveric human donors (h NHBD) processed within 24 h, fresh porcine donors (p-HBD) and cadaveric porcine donors (p-NHBD) also processed within 24 h. Leaflets were separately incubated at 37 degrees C for successive periods of 30 min up to 5 h in Earle's Medium 199. After 240 min PGI-2 production was stimulated by 10 microM bradykinin. Postincubational release was stopped with indomethacin 10 microg/ml. Prostacyclin production was measured as 6-kPGF1a using an ELISA. RESULTS: Initial PGI-2 production is significantly higher in porcine than in human grafts and in both species enhanced by previous warm ischemia. While baseline species differences disappear during progressive incubation, differences resulting from graft history are maintained. After PGI-2 stimulation species differences dominate again while ischemic history has no effect. CONCLUSION: Ischemia and surgical manipulation are stimulators of endothelial PGI-2 production in both human and porcine allografts and, therefore, a correlation of this metabolic activity with cellular integrity may be misleading. Valid data are obtained only if the natural time course and reaction to stimulation of PGI-2 production are duely recognized and species differences in the response to mechanical and ischemic stress are considered. PMID- 9860208 TI - Hypothermic preservation of isolated rat lungs in modified bicarbonate buffer, EuroCollins solution or St Thomas' Hospital cardioplegic solution. AB - OBJECTIVES: Inadequate preservation solutions limit lung storage times and, consequently, transplant programs. To address this problem we established an isolated, ventilated and perfused rat lung preparation. Here we report the effects of hypothermic storage in EuroCollins solution, St Thomas' Hospital cardioplegic solution and a modified bicarbonate buffer solution. METHODS: Lungs from male Wistar rats (230-330 g) were perfused via the pulmonary artery with modified bicarbonate buffer (37 degrees C, 15 ml/min, constant flow) and ventilated by positive pressure (tidal volume:1.6-1.8 ml, 80 breaths/min). Vascular resistance (pulmonary artery pressure:perfusate flow ratio) and airways compliance (tidal volume:tracheal pressure ratio) were measured. After a control perfusion period (20 min), lungs were flushed with, then immersed in, bicarbonate buffer (4 degrees C) for varying periods (0-24 h). After storage, lung function was assessed during 20 min reperfusion. Having established a suitable period for study, storage in EuroCollins, St Thomas' Hospital cardioplegic solution or bicarbonate buffer were compared. RESULTS: Pulmonary compliance (ml/cmH2O) was significantly (P < 0.05) reduced in lungs stored for 6 h in modified bicarbonate buffer (0.026 +/- 0.008), EuroCollins solution (0.013 +/- 0.002) or St Thomas' Hospital solution (0.025 +/- 0.005) compared to unstored lungs (0.068 +/- 0.007). Vascular resistance, (1.32 +/- 0.13 cmH2O/ml per min) in unstored lungs, was similar in lungs stored in St Thomas' Hospital solution but increased significantly in lungs stored in modified bicarbonate buffer (3.22 +/- 0.78 cmH2O/ml per min) or EuroCollins solution (4.66 +/- 0.57 cmH2O/ml per min). CONCLUSIONS: Hypothermic storage of rat lungs for 6 h in modified bicarbonate buffer or St Thomas' Hospital solution causes less increase in vascular resistance on reperfusion than EuroCollins solution. PMID- 9860209 TI - Images in cardio-thoracic surgery. Caught in the act. PMID- 9860210 TI - Isolated pericardial rupture with left-sided haematothorax after blunt chest trauma. AB - Pericardial rupture after blunt chest trauma is described in the literature. The rupture is mostly caused by high velocity trauma with associated injuries. As a result the patients are often critically ill. We describe a case of a 59-year-old man who suffered from an isolated pericardial rupture with a left-sided haematothorax diagnosed 3 months after minimal blunt chest trauma. The patient was operated upon and the defect was closed without detrimental sequelae. PMID- 9860211 TI - A case of giant chondrosarcoma of the cricoid cartilage presenting as a superior mediastinal tumour. AB - We report the case of a 73-year-old man operated on for a symptomatic superior mediastinal mass which developed from the cricoid cartilage. Through cervicotomy and partial sternotomy, laryngeal subglottic resection allowed complete removal of the tumour. From pathological examination, a benign chondroma was initially diagnosed without any criterion of malignancy. However, subsequent review of the pathological slides showed the lesion to be a low grade chondrosarcoma with an epithelial quota of spindle cells, responsible for the patient's death. This case illustrates the need for resection of presumed cricoid cartilage chondromas because of their potential mediastinal development and associated malignancies. PMID- 9860212 TI - Primary angiosarcomas of the chest wall and pleura. AB - Primary angiosarcomas of the chest wall and pleura are extremely rare and carry a dismal prognosis. Two cases are reported. One patient (case 1), presented with massive recurrent haemothorax, was found to have multifocal angiosarcoma of the pleura, treated with surgical de-bulking, chemical pleurodesis and chemotherapy, achieving control of the bleeding. She died 10 months later from complications related to chemotherapy. A full post-mortem examination confirmed this was a primary pleural angiosarcoma with no evidence of disease elsewhere. Another patient (case 2) with a large solitary angiosarcoma of the chest wall, discovered incidentally on a routine physical examination, was successfully treated with surgical excision and subsequent radical radiotherapy, remaining well 15 years post-operatively. PMID- 9860213 TI - Cardiopulmonary bypass in sickle cell anaemia without exchange transfusion. AB - To minimize sickling during cardiopulmonary bypass, exchange transfusion is frequently recommended peri-operatively for patients with homozygous sickle cell anaemia to reduce the circulating concentration of HbS. We report the successful management of two children both aged 12 years with sickle cell anaemia (homozygous SS), several sickle cell crises and multiple blood transfusions who underwent cardiopulmonary bypass for mitral valve replacement. No pre- or intra operative exchange transfusions were employed in the two cases. Furthermore, moderate hypothermia in both cases was not associated with adverse sequelae. PMID- 9860214 TI - Aspergillus endocarditis: rare but serious Aspergillus ball obstructing the pulmonary artery. AB - Aspergillus endocarditis is considered a rare complication in cardiac surgery, which is usually fatal due to unclear clinical manifestations. We present two cases with early and late post-operative Aspergillus endocarditis. Early diagnosis, more investigation for appropriate antifungal agents and their dosage, and early and aggressive surgical treatment may decrease fatality. PMID- 9860215 TI - Infective endocarditis complicated by mycotic cerebral aneurysm: two case reports of women in the peripartum period. AB - Mycotic cerebral aneurysm is a relatively rare but very serious complication of infective endocarditis. Infective endocarditis is a rare but a potentially fatal complication of pregnancy. We report here two very rare cases of infective endocarditis associated with mycotic cerebral aneurysm in peripartum women. In one case, cardiac surgery was performed prior to cerebral surgery and after delivery. In the other case, emergency cerebral surgery was performed due to rupture on the day cardiac surgery had been scheduled, 45 days after delivery. The surgical management of a patient with infective endocarditis and mycotic cerebral aneurysm is reviewed. The surgical strategy for a pregnant patient is also reviewed. PMID- 9860216 TI - Right ventricular rupture in minimally invasive direct coronary artery bypass grafting. AB - We report a rare case of the rupture of the right ventricle which occurred in a minimally invasive direct coronary artery bypass grafting (MIDCABG) for a redo bypass surgery. A 52-year-old male patient underwent a left internal thoracic artery (LITA) to the left anterior descending artery (LAD) bypass. Rupture of the right ventricle occurred abruptly during dissection to find the LAD. Too much dissection of the interventricular groove under undue traction of the pericardium may cause a rupture of the heart. PMID- 9860217 TI - Metastatic malignant melanoma as an intracavitary obstructive mass in the right heart. PMID- 9860218 TI - Cardiovascular syphilis: a rare medical diagnosis? PMID- 9860219 TI - Basic surgical research. PMID- 9860220 TI - Clinical research in surgery: questions but few answers. AB - BACKGROUND: Surgeons must be prepared to be questioned about their understanding, activities, and achievements in basic and clinical research in terms of structural, organizational, and financial support of surgical research. RESULTS: Clinical research today comprises basic research related to accumulation of knowledge about biological systems, disease-related research including experimental research with animals, and in vitro research with biological material and clinical studies with the strongest empirical basis in data derived from controlled clinical trials. Most clinical methods related to diagnosis and treatment have been introduced into the practice of everyday medicine even in university hospitals without passing a prospective evaluation protocol. To test the efficacy of a new drug or a newly developed surgical technique, the controlled prospective clinical trial is the best method; however, fewer than 50% of clinical questions can be answered by controlled clinical studies. In Germany the frequency of controlled clinical trials for answering clinical questions is presently low; this is particularly true for surgery, in comparison with other countries. The concept of evidence-based medicine is based on the principles of data generation from controlled trials and meta-analyses of those studies. Best evidence synthesis means a reasonable synthesis of evident knowledge, experience and intuition. The challenge of professionalization of clinical research in surgery is linked to the establishment of full time positions for basic scientists and clinical researchers and the establishment of clinical research teams and of research professorships including basic scientists in surgical departments. CONCLUSION: Clinical research can be effective only if there is good cooperation between clinicians and basic scientists, if high grade scientific methods are established in clinical research institutions, if there is sufficient financial and personal supply, and if there are enough laboratories and animal operating facilities. There is an urgent need for structures integrating basic scientists in clinical research groups, for more interdisciplinary research projects and more multidisciplinary research, independent of clinical care in terms of manpower. PMID- 9860221 TI - Conceptional and structural conditions for successful clinical research. AB - The efficiency of surgical research has again become the subject of debate. Clinical research is required to improve our understanding of surgical disorders and our ability to treat patients. This involves both experimental research (research in the test tube) and clinical research in actual patients. The surgeon must remain the expert because it is he who deals with the patient and is confronted with his problems. On the other hand, care for the patient, must always be the central issue. Here a new orientation is needed, evaluating the effectiveness of surgical research from the patient's point of view. Surgical treatment, particularly surgical research, must be adapted first to the individual patient and only secondly to the surgical disease - the problem must determine the method, not vice versa. While it is clear that a creative atmosphere, supportive structure and efficient organisation are enormously helpful, today's exaggerated attention to matters of structure and organisation are often poor substitutes for creativity and intuition. Surgical research does not refer solely to therapy research but includes methods for carrying out controlled clinical trials, establishing guidelines and scores and designing instruments for measuring outcome. Socioeconomic and analyses and ethical considerations are crucial for facing such conflicts as "quality versus quantity", "profession versus business", "patient care versus economics costs". Proposals for designing more effective concepts, structure and organisation for clinical research are presented here, and three models are introduced: the cooperation model, the integration model and a mixture of the two. PMID- 9860222 TI - Gastroesophageal reflux in neurologically impaired children: partial or total fundoplication? AB - BACKGROUND: It is difficult to give guidelines when approaching gastroesophageal disease in neurologically impaired children. Indication for surgery has been increasing over recent years, but there is no consensus on the surgical technique of choice. Nothing has been written specifically comparing the results of different procedures in these patients, so far. STUDY DESIGN: We retrospectively compare the short- and long-term results of two different types of fundoplication in a series of children operated on for documented gastroesophageal reflux disease at our institution. RESULTS: One group (group A) of 27 patients, operated on between 1977 and 1993, underwent Nissen fundoplication, the other (group B), formed of 20 patients all of whom were operated on between 1993 and 1995, underwent Thai fundoplication. We compared the results in terms of positive outcome (recovery) and negative outcome (minor and major complication), computing the relative odds of group A versus group B in terms of risk of complication, and we compared the mean operative time and the length of hospital stay by means of a student's t-test analysis. CONCLUSIONS: Our results show that there is no statistical difference between the two procedures in terms of relative risk of complication and success rate. The duration of surgery and hospital stay were significantly shorter in group B. The Thal procedure can, therefore, be proposed as first choice in the management of these patients. PMID- 9860223 TI - Follow-up after transanal endoscopic microsurgery or transanal excision of large benign rectal polyps. AB - METHODS: Between January 1986 and December 1995, 238 patients with benign rectal polyps underwent either transanal endoscopic microsurgery (n = 226) or transanal excision (n = 12) at the Clinic of General and Abdominal Surgery, Johannes Gutenberg-University, Mainz. RESULTS: Mean polyp size was 4.2 cm; 89.1% of polyps measured more than 2 cm in diameter. In 89.1% of cases, histological analysis revealed polyps containing tubulovillous or villous adenomas. Synchronous colonic polyps were detected in 12.5% of patients. Follow-up data are available on 222 patients (94%). At follow-up examination, 169 of the 193 surviving patients (87.6%) were recurrence free. Seven of 193 patients (3.6%) had developed neoplastic colonic polyps and, in 17 patients (8.8%), metachronous polyps were detected. CONCLUSIONS: Transanal endoscopic microsurgical polypectomy was furthermore demonstrated to be a low-risk procedure with a low recurrence rate for the complete resection of large rectal polyps. At a follow-up rate of 61.1 %, the incidence of metachronous carcinoma ranged at 3.1%, which is markedly below the rate of 8-18% for tubulovillous or villous adenomas larger than 1 cm in diameter cited in the literature. PMID- 9860224 TI - Inter- and intraindividual reproducibility of anorectal manometry. AB - BACKGROUND: This study investigates the inter- and intraindividual variability of normal values and, thus, the reproducibility of anorectal manometry. MATERIALS AND METHODS: Following a standardized protocol, three anorectal manometries were performed 4 h apart on 2 days of investigation, with an interval of 4 weeks, in ten healthy volunteers. Measured parameters in all 60 manometries were: sphincter length (SL), resting pressure (RP), maximum squeeze pressure (MSP), relaxation of the internal anal sphincter (RIAS), and rectal compliance (RC). Interindividual variability was expressed as standard deviation from calculated mean values and intraindividual variability was tested with Wilcoxon's test for tied samples and Spearman's rank correlation test. RESULTS: A large interindividual variability was found for all measured parameters, except for SL, reflecting the extensive absolute range of measured values. Median intraindividual variability among the six individual measurements and between both measurement days revealed that MSP, RIAS and RC are parameters which were not reproducible in this volunteer study. A significant correlation between the results of the repetitive measurements and, thus, a good reproducibility was only found for the parameters SL and RP. CONCLUSIONS: Anorectal manometry has only limited diagnostic value; although rather exact quantifications of individual parameters can be achieved, the impact of these measurements should be regarded rather critically, since only SL and RP appeared to be reproducible parameters. PMID- 9860225 TI - Experience gained from operation of 103 adrenal incidentalomas. AB - BACKGROUND: Adrenal incidentaloma poses a perplexing problem for physicians. The challenge is to identify the rare functioning or malignant adrenal tumor that should be removed. PATIENTS AND METHODS: From 1970 until December 1996, we managed 579 patients with adrenal hyperplasia or neoplasm. Of these, 110 were followed without any operation and 469 were operated on. The histology from the operated patients revealed 145 pheochromocytomas, 72 Cushing's syndrome, 98 aldosteronomas, 21 virilizing tumors, 3 feminizing tumors, 98 non-functioning adrenalomas, 17 metastases, 7 cysts, and 15 miscellaneous tumors. RESULTS: Among the 98 non-functioning adrenal tumors, 79 were incidentalomas. Histological examination of 103 operated incidentalomas found 55 adenomas (5 functioning=aldosteronomas), 12 benign pheochromocytomas, 3 malignant or suspicious pheochromocytomas, 8 cystic lymphangiomas, 4 metastases, 4 angiomas, 3 ganglioneuromas, 4 myelolipomas, 1 hyperplasia, 2 hematomas, 2 cysts, and 5 adreno-cortical carcinomas. Among the non-operated incidentalomas, the most frequent findings were adenomas, metastases, hematomas, cysts and pseudotumoral hyperplasia; three disappeared spontaneously. No patients died of unrecognized adrenocortical carcinoma. CONCLUSION: Our experience confirms that in case of incidentaloma, the first concern is a nonfunctional pheochromocytoma (15%), and the less common occurrence is adreno-cortical carcinoma (2%). We recommend a restrictive surgical approach in the setting of adrenal incidentalomas. PMID- 9860226 TI - German medullary thyroid carcinoma/multiple endocrine neoplasia registry. German MTC/MEN Study Group. Medullary Thyroid Carcinoma/Multiple Endocrine Neoplasia Type 2. AB - INTRODUCTION: A national registry for medullary thyroid carcinoma (MTC) and multiple endocrine neoplasia type 2 (MEN2) was set up to evaluate epidemiological, clinical and prognostic factors of the sporadic and hereditary forms of MTC. PATIENTS AND METHODS: Coded data from 1217 patients with MTC from 57 participating centers in Germany were registered and statistically analyzed. The series included 515 (42%) males (mean age 43.1+/-16.1 years) and 702 (58%) females (mean age 44.4+/-17.6 years), with a mean follow-up of 5.2 years; 865 (71%) exhibited the sporadic form and 352 (29%) the familial form (244 MEN2a, 32 MEN2b, 76 FMTC). The mean age at diagnosis was 49 years for sporadic and 30 years for the familial form. RESULTS: Of the patients, 17% presented at stage I, according to the UICC, and 30%, 45%, and 8% presented at stages II, III, and IV, respectively. There were 12% of patients who died of the disease. The overall adjusted survival rate was 87% at 5 years, and 76% at 10 years. In a univariate analysis, the stage of disease at diagnosis, age, gender, and form were relevant prognostic factors. In a multivariate proportional-hazards analysis, the difference between patients with sporadic and familial disease disappeared. CONCLUSION: As the tumor stage at presentation is the major prognostic factor, early diagnosis and surgical intervention before cervical lymph node metastases appear is necessary to improve survival especially in sporadic cases. PMID- 9860227 TI - Thyroid carcinoma in intrathoracic goiter. AB - INTRODUCTION: Most cases of intrathoracic goiter can be managed by cervical incision alone. A thoracic approach may be needed when adhesions or an anomalous blood supply are present or carcinoma is suspected. PATIENTS AND METHODS: Only 44 patients out of 5263 operated on for goiter needed a thoracic incision. A sternotomy was performed in 29 cases and a thoracotomy in 15; a malignancy was present in 9 cases. Symptoms, surgical approach, histology, survival and pTN staging of these 9 patients were reviewed and discussed; no perioperative mortality was observed. DISCUSSION: A thoracic approach is more frequently needed for treatment of intrathoracic thyroid carcinoma as it offers a greater chance of radical excision and better control of intraoperative bleeding. Histologically, thyroid carcinoma in intrathoracic goiter is often anaplastic or rare and has a poor long-term survival rate when compared to cervical forms. PMID- 9860228 TI - Polysialylated neural cell adhesion molecule in childhood ganglioneuroma and neuroblastoma of different histological grade and clinical stage. AB - BACKGROUND AND AIMS: Neuroblastoma cells express the polysialylated form of the neural cell adhesion molecule (PSA-NCAM), which normally becomes restricted to a few neural regions after embryogenesis. The aim of the present study was to evaluate PSA-NCAM as a marker in childhood neuroblastoma. PATIENTS/METHODS: We studied the expression of PSA-NCAM on tumor specimens and in sera of 27 children, altogether, with ganglioneuroma and neuroblastoma of different histological grades and clinical stages. For both methods, immunohistochemistry on 5-microm frozen sections and immunoluminescence serum assay, the polysialic-acid-specific monoclonal antibody 735 was used. RESULTS: PSA-NCAM expression was highest in patients with undifferentiated neuroblastoma and advanced stages of disease, whereas children with differentiated tumor types and low clinical stages had distinctly reduced or no reactivity in immunohistochemistry and, simultaneously, normal serum levels. PSA-NCAM expression correlated with other prognostic and diagnostic markers, such as MYCN gene amplification, and serum concentrations decreased during successful treatment. CONCLUSIONS: We conclude that PSA-NCAM, both immunohistochemically and in the serum, is a promising candidate for another useful diagnostic and prognostic tumor marker in childhood neuroblastoma. PMID- 9860230 TI - Effect of granulocytes on the microcirculation in free-flap surgery. AB - INTRODUCTION: Impaired capillary perfusion may result in flap failure. Platelet emboli, vasospasm and/or polymorphonuclear leukocytes (PMNs) have been identified as possible causes. This study investigates the role of PMNs in causing impaired capillary perfusion in a free-flap model. METHODS: Sprague-Dawley rats were injected with either anti-neutrophil serum or saline. Their cremaster muscle was isolated on its pedicle. After arterial repair and reperfusion, capillary perfusion was counted each hour for 6 h. RESULTS: The number of PMNs was significantly reduced in the animals treated with anti-neutrophil serum. However, capillary perfusion did not improve in this group. CONCLUSIONS: These results demonstrate that depleting circulating PMNs does not counterbalance the reduction of perfused capillaries, i.e., does not increase their number. It is suggested that reduced capillary perfusion downstream from an anastomotic repair is not mediated by the presence of PMNs in the microcirculation. PMID- 9860229 TI - Genetically modified fibroblasts induce angiogenesis in the rat epigastric island flap. AB - METHODS: Gene therapy was tested for inducing functional angiogenesis in the superficial rat epigastric island flap to allow earlier pedicle division. Autologous rat fibroblasts were grown, harvested, cultured and retrovirally transfected to produce platelet-derived growth factor AA (PDGF-AA), an angiogenetically active protein. Stable gene expression was monitored by PDGF-AA enzyme-linked immunosorbent assay (ELISA). One hundred and eighty animals were divided into three groups (I-III) and a bilateral flap created in each animal. In all experiments, the right-sided flap was subjected to experimental treatment and the left-sided flap served as control (1ml saline 0.9%). During flap elevation, group I received 5X10(6) GMFB (genetically modified fibroblasts) plus 1 ml Dulbecco's modified Eagle's medium. Group II was treated with 5x10(6) NMFB (non modified fibroblasts) plus 1 ml medium and group III received 1 ml medium only. The flaps were sutured back and the vascular pedicle was bilaterally ligated and divided in each of ten animals during the following 6 days. After 7 days, the flaps were harvested, the amount of necrosis measured and histologically examined. RESULTS: The GMFB produced up to 560 times more PDGF-AA than the NMFB, measured by ELISA. The GMFB-treated flaps tolerated surgical division of the vascular pedicle significantly earlier than groups II and III. Histologically, fibroblasts persisted in all flaps of groups I and II, without major inflammatory reaction. In all GMFB-treated flaps, massive angiogenesis could be demonstrated. CONCLUSION: By means of retroviral gene transfer, autologous rat fibroblasts can be genetically modified for stable expression of the PDGF-A gene to produce high amounts of PDGF-AA, which is angiogenetically active. After injection into the panniculus carnosus, these cells induce functional angiogenesis to permit earlier division of the vascular pedicle in this flap model. PMID- 9860231 TI - Lymph node dissection in patients with differentiated thyroid carcinoma--who benefits? AB - INTRODUCTION: Papillary and follicular thyroid carcinomas are the most common thyroid malignancies and are usually indolent. Lymph-node involvement increases the rate of tumor recurrence and reduces long-term survival. However, characteristics such as age, long-term iodine deficiency, histological grade, extrathyroidal extension and distant metastases are clearly much more potent adverse factors than cervical lymph node metastases. Although there are no prospective randomised studies showing the overall benefit of routine cervicocentral lymph-node dissection in addition to total thyroidectomy, we propose this procedure as a standard part of primary surgery in differentiated thyroid carcinoma (DTC). RESULTS: In specialised centres, morbidity is not higher than for thyroidectomy alone. However, there is a significant increase in morbidity after re-operation in the cervicocentral compartment. Modified neck dissection of the cervicolateral compartment should only be performed if there is clinical evidence of lymph-node involvement in this area. CONCLUSIONS: This strategy provides the optimal surgical treatment for all subgroups of patients with DTC and creates optimal conditions for effective postoperative radioiodine ablation, which is another prognostic factor strongly associated with recurrence and survival in these patients. PMID- 9860232 TI - Cost analysis of three different surgical procedures for treatment of a pelvic tumour. AB - INTRODUCTION: Malignant pelvic tumours are rare, but adequate treatment is difficult because of anatomical and functional reasons. Different surgical procedures are recommended. Besides aspects influencing the quality of life due to the different surgical techniques, costs of these surgical procedures and the perioperative time interval, depending on the chosen surgical procedure, must also be considered. METHODS: Costs of three kinds of surgical treatment - internal hemipelvectomy vs external hemipelvectomy and application of an orthotic device vs ilio-femoral pseudoarthrosis - were compared, including costs of the immediate period of rehabilitation. Costs of the preoperative diagnostic procedure were excluded because they were assumed to be equal. For all calculations, treatment of a periacetabular pelvic tumour type II-b was assumed, according to the Enneking classification, with the need for a pelvic resection with wide margins including removal of the hipjoint. This analysis was performed based on the average costs per hour of physicians, nurses and physiotherapists, including a basic daily rate for additional costs, such as housekeeping and administrative costs of the hospital. In all cases, the costs of the operative procedure and the perioperative period up to 3 months were calculated, including the costs for the endoprosthetic device after internal hemipelvectomy and those for the orthotic devices after necessary external hemipelvectomy. RESULTS: For a postoperative period of 3 months, the cost for treatment with resection of the tumour and performing an ilio-femoral pseudoarthrosis is nearly DM 56,741.54; treatment with an internal hemipelvectomy including the endoprosthetic pelvic replacement costs DM 81,439.34; and treatment with external hemipelvectomy and application of an orthotic device with a pelvic cage costs DM 69,138.46. In this analysis, social costs due to the different rates of disability for years or costs of a new prosthetic device after years and aspects related to resulting quality of life were excluded. In contrast, costs of three different actual cases can differ significantly from the estimated costs. The most important influencing items are surgical implants, blood units and similar products, and surgical treatment of postoperative complications. CONCLUSIONS: Regarding all these aspects, limb salvage seems to be advantageous over amputation. Whether an ileo femoral pseudoarthrosis or endoprosthetic pelvic replacement should be performed is a decision that should be made by the physicians. based on the underlying diagnosis and the correlated expected survival. Furthermore, additional factors probably reducing the rate of postoperative complications, such as infections or necrosis following radiotherapy and being summarised under the term "quality of life", should also be regarded. PMID- 9860233 TI - Allergy--atopy--hypersensitivity--a matter of definition. PMID- 9860234 TI - Cow's milk allergens. PMID- 9860235 TI - Neutral endopeptidase 24.11: its physiologic and possibly pathophysiologic role in inflammation with special effect on respiratory inflammation. PMID- 9860236 TI - IgE antibodies specific for carbohydrates in a patient allergic to gum arabic (Acacia senegal). AB - The present study deals with the detailed investigation of the IgE antibody response of a gum arabic-allergic patient. The patient showed multiple serologic and skin test sensitizations to a range of pollen, other inhalants and foods, and bee venom, and to the recombinant allergens Bet v 1 and Bet v 2. Moreover, the patient's serum reacted strongly to gum-arabic extract. The NaIO4-treated and thus deglycosylated extract showed no binding to IgE. In contrast, removal of the protein backbone by basic hydrolysis did not deplete the IgE reactivity. Therefore, it is concluded that the gum arabic-specific IgE antibodies of this patient were mainly directed against the carbohydrate fraction of this material. In IgE-inhibition assays, cross-reactions occurred in the range of 60% between gum arabic and known immunogenic N-glycans containing alpha1-3-linked fucose. Since the inhibition graphs were not parallel and the inhibition was not complete with heterologue antigens, the cross-reacting epitopes of gum arabic appeared to be different from the latter well-known cross-reactive carbohydrate determinants (CCD). Inhibition may have been caused by a partial immunologic identity of the investigated carbohydrate moieties. A strong IgE response to the fucose containing glycan from bromelain was measured in a glycan ELISA that utilizes purified glycopeptides at the solid phase. This response, which may explain the multiple sensitizations without clinical significance diagnosed in the patient, could originate from inhalation of pollen, which is known to contain similar glycans, or from occupational sensitization during work as a baker and confectioner. Since the gum-arabic protein showed only very weak participation in the IgE reactivity, the clinical symptoms of the patient caused by gum arabic may be attributed to carbohydrate epitopes. Due to the repetitive polysaccharide sequence of gum arabic, several epitopes for the cross-linking of IgE should exist. PMID- 9860237 TI - In vitro effects of the pyrethroid S-bioallethrin on lymphocytes and basophils from atopic and nonatopic subjects. AB - Synthetic pyrethroids are increasingly used as insecticides and marketed as having relatively low human toxicity. The aim of this study was to examine the in vitro effects of the synthetic pyrethroid S-bioallethrin on human blood lymphocytes and basophils in atopic individuals and nonatopic control subjects. S bioallethrin caused inhibition of lymphocyte proliferation after a 72-h culture period in a concentration-dependent manner. The inhibition of the lymphocyte proliferation by S-bioallethrin at the concentration 6.5 microM correlated well with the total serum IgE values (r = -0.89, P < 0.001). Samples from atopic subjects were more sensitive to this inhibition than those from nonatopic volunteers. The regulatory interleukin-4/interferon-gamma (JL-4/IFN-gamma) balance showed a significant difference between atopic and nonatopic subjects after a short-term culture period (24 h) in the presence of the same concentration range of S-bioallethrin (P < 0.001). Additionally, IFN-gamma secretion was consistently lower in cells from the atopic donors. Furthermore, S bioallethrin induced histamine release from human basophils in a concentration dependent manner. Although the effect was small compared to histamine liberators such as N-formyl-Met-Leu-Phe and anti-IgE, the response to S-bioallethrin was significantly different in atopic donors from nonatopic (P = 0.0431). These findings are the first demonstration of the immunotoxicologic properties of the synthetic pyrethroid S-bioallethrin by this combined in vitro approach with human lymphocytes and basophils. Further studies will investigate the responses of lymphocytes from patients who are sensitive to these agents. PMID- 9860238 TI - Indoor environmental factors associated with house-dust-mite allergen (Der p 1) levels in south-eastern Australian houses. AB - BACKGROUND: Eighty households in the Latrobe Valley, Victoria, Australia, were sampled for house-dust-mite allergen (Der p 1). Allergen levels vary greatly between houses within climate regions. The reasons for this are not well understood. METHODS: House-dust-mite allergen samples were collected on six occasions between March 1994 and February 1995. All participating households contained at least one child between 7 and 14 years with a total of 148 subjects, 53 of whom were asthmatic. A detailed house survey was performed during every sampling visit, and a dwelling questionnaire was completed. Relative humidity was measured at the time of sample collection. RESULTS: The median bed allergen level was 30 microg/g during the first sampling period. Significantly higher allergen levels were associated with wool bedding and inner-spring mattresses (P < 0.001). As estimated from a multiple linear regression model, up to 70% reduction in bed allergen levels may be achieved by avoiding wool bedding and inner-spring mattresses. Other risk factors for high allergen levels included high indoor relative humidity, presence of substantial visible mould growth, brick cladding, and concrete slab foundation of the house. CONCLUSIONS: Avoiding wool bedding and replacing inner-spring mattresses with foam could substantially reduce bed allergen levels. PMID- 9860240 TI - Role of nonallergic hypersensitivity reactions in children with chronic urticaria. AB - BACKGROUND: IgE-independent (pseudoallergic) reactions to food and food ingredients are common in a subgroup of adult patients with chronic urticaria, who have daily spontaneous occurrence of wheals. However, for children with chronic urticaria (duration longer than 6 weeks, no physical influence), no data on the importance of pseudoallergen-induced chronic urticaria are available. Therefore, we investigated the role of nonallergic hypersensitivity to food in all children seen with chronic continuous urticaria in our two clinics over the last 2 years (n = 16). METHODS: All patients were given a low-pseudoallergen diet for 3 weeks followed by provocation with food rich in pseudoallergens. To identify the main eliciting agents, a subgroup of responders was exposed to food additives by double-blind, placebo-controlled food challenges. RESULTS: Pseudoallergen-induced urticaria was diagnosed in 12 cases (75%). Reactions occurred mainly to coloring agents and preservatives, but also to monosodium glutamate and a sweetener (saccharin/cyclamate). CONCLUSIONS: These results confirm that nonallergic hypersensitivity reactions play a role in children with chronic urticaria, although the latter disease is rare at that age. In children, food additives, especially coloring agents and preservatives, appear to play a more important role in eliciting nonallergic hypersensitivity reactions than in adult patients, where naturally occurring pseudoallergens in fruits and vegetables are mainly responsible. PMID- 9860239 TI - Eosinophil leukocyte degranulation in response to serum-opsonized beads: C5a and platelet-activating factor enhance ECP release, with roles for protein kinases A and C. AB - BACKGROUND: Eosinophils have a typical content of granule-bound, cytotoxic, cationic proteins which may, when released to the external milieu, play roles in diseases such as asthma and parasitic infestation. Therefore, we have investigated possible mechanisms by which their release is regulated in eosinophils. METHODS: The enzyme-linked immunosorbent assay (ELISA) was used to detect released eosinophil cationic protein (ECP). Release of ECP was induced by serum-opsonized, nonphagocytosable Sephadex beads (SOS). RESULTS: The complement fragment C5a and platelet-activating factor (PAF) were found to enhance ECP release in response to SOS in a dose-dependent fashion, and, contrary to previous reports, they were not found to act as secretagogues themselves on eosinophils in suspension. The role of protein kinase C (PKC) in eosinophil degranulation has been controversial. We found that ECP release induced by SOS was inhibited by the PKC inhibitors staurosporine and calphostin C. Activation of protein kinase A (PKA), by raising cAMP, also inhibited ECP release. Furthermore, pertussis toxin decreased ECP release on opsonized beads, indicating the involvement of pertussis toxin-sensitive G proteins. CONCLUSIONS: C5a, and PAF enhance granule release, rather than acting as secretagogues themselves. PKC and PKA have opposing roles in the regulation of ECP release in response to SOS. PMID- 9860241 TI - Contact urticaria from protein hydrolysates in hair conditioners. AB - Protein hydrolysates (PHs) are added to hair-care products (to "repair" broken hair), soaps, bath gels, creams, etc. From one to 22 PHs used in hair-care products (collagen, keratin, elastin, milk, wheat, almond, and silk) were tested in three patient groups: A) 11 hairdressers with hand dermatitis B) 2160 consecutive adults with suspected allergic respiratory disease subjected to routine skin prick tests C) 28 adults with atopic dermatitis. In group A, all the 22 PHs were tested with scratch and patch tests. In groups B and C, one to three PHs were tested with prick tests. Positive scratch/prick test reactions were seen in 12 patients from three PHs altogether. All were women with atopic dermatitis, and all reacted to at least hydroxypropyl trimonium hydrolyzed collagen (Crotein Q). In three patients, prick and open tests with a hair conditioner containing Crotein Q were performed with positive results. One patient reported contact urticaria on her hands, and two reported acute urticaria on their head, face, and upper body from a hair conditioner containing Crotein Q. In seven of the eight studied sera, specific IgE to Crotein Q was detected. In conclusion, PHs of hair cosmetics can cause contact urticaria, especially in patients with atopic dermatitis. PMID- 9860242 TI - Lack of association between atopy and RsaI polymorphism within intron 2 of the Fc(epsilon)RI-beta gene in a Spanish population sample. AB - BACKGROUND: The B genotype of RasI polymorphism located within intron 2 of the Fc IgE receptor I (Fc(epsilon)RI) gene was previously found to be increased in atopic patients from a Japanese population sample. METHODS: We studied these A/B genotypes in 70 Spanish atopic patients, and the results were compared to those of 51 nonatopic controls. RasI polymorphisms were studied by specific digestion of polymerase chain reaction fragments followed by polyacrylamide gel electrophoresis. RESULTS: The polymorphism frequency (A/A: 25/70, A/B: 28/70, B/B: 17/70) found in patients did not differ from the frequency in nonatopic control subjects. CONCLUSIONS: We did not find RasI polymorphisms associated with atopic disease. The genetic findings in atopy and asthma may be very different according to ethnic and local characteristics, and they must be carefully verified in different population samples. PMID- 9860243 TI - Diagnostic tests in children with atopic dermatitis and food allergy. AB - BACKGROUND: Skin testing is a common diagnostic procedure in food allergy. The skin prick test is the test of first choice for investigating the immediate IgE mediated reaction. The skin application food test (SAFT) has been developed on the basis of the mechanism of the contact urticaria syndrome (CUS). METHODS: We studied the relevance of the SAFT in children younger than 4 years with atopic dermatitis and (suspected) food allergy as compared with the prick-prick test, the radioallergosorbent test (RAST), and the oral challenge. In the skin tests, we used fresh food, in the same state as it was consumed. RESULTS: There was a good agreement between the SAFT and the prick-prick test. A moderate agreement was observed between the SAFT and the serologic test (RAST). Significantly more positive results in the RAST were observed than in the SAFT. There was very good agreement between the SAFT and the oral challenge (kappa = 0.86). CONCLUSIONS: The SAFT is a reliable and child-friendly skin test for evaluating (suspected) food allergy in children younger than 4 years with atopic dermatitis. The very good correlation with the oral challenge indicates that one may probably consider the SAFT a "skin provocation" in children younger than 4 years. PMID- 9860244 TI - Alveolar macrophage interleukin (IL)-10 and IL-12 production in atopic asthma. AB - Inflammation in asthma is characterized by a Th2 response. In many experimental systems, this response can be regulated by interleukin (IL)-10 and IL-12. IL-10 deactivates T cells, and IL-12 reorients the response toward a Th1 pattern. Alveolar macrophages (AM) can secrete both of these cytokines, and thus regulate T-cell behavior in asthma. They can enhance the Th2 response by turning off their secretion of IL-10 and IL-12, or tend to downregulate it by producing these cytokines. To elucidate that point, we assayed the AM IL-10 and IL-12 from 11 asthmatic patients and four controls. Six asthmatics were treated by inhaled corticosteroids. AM were recovered by bronchoalveolar lavage (BAL). They were isolated and cultured for 24 h without stimulation or in the presence of lipopolysaccharide (LPS). IL-10 and the p40 subunit of IL-12 were assayed in the BAL fluid and in AM culture supernatants by ELISA. Spontaneous AM IL-10 production was higher in asthmatics, particularly in the treated group. The AM IL 10 production after stimulation by LPS was also elevated in asthmatics, but was mainly so in untreated patients. IL-12 levels were higher in BAL fluids from untreated patients than from controls. The IL-12 production of LPS-stimulated-AM from these patients was increased. These results show that AM are at least primed for the production of IL-10 and IL-12 in asthma, and suggest that these cells could be involved in the resolution of the asthmatic inflammation. PMID- 9860245 TI - Prevalence of occupational asthma among automobile and furniture painters in the center of Eskisehir (Turkey): the effects of atopy and smoking habits on occupational asthma. AB - BACKGROUND: Occupational asthma (OA) is a respiratory disorder characterized by airway hyperreactivity caused by agents present in the workplace. For determination of the prevalence of OA among car and furniture painters exposed to isocyanate in the center of Eskisehir, Turkey, a clinical and epidemiologic prospective study in three phases was done, incorporating 312 (89.4%) of the painters. METHODS: Of these subjects, 190 (61%) were furniture painters and 122 (39%) automobile painters. In the first phase of the study, a modified questionnaire and pulmonary function test (PFT) were done. During the second phase, peak expiratory flow rate (PEFR) was monitored in 52 subjects whose complaints were confirmed and who agreed to a month of such monitoring. In the third phase, nonspecific bronchial provocation tests (NSBPT) with histamine were done on 23 of the PEFR-monitored workers. RESULTS: Finally, through questionnaire, typical history, PFT, PEFR monitoring, and NSBPT, 30 workers (9.6%) were diagnosed as having OA. Smoking habits and atopy in the OA-diagnosed workers were found to be statistically significantly high in comparison to the other workers. CONCLUSIONS: It was concluded that OA is a common disorder among automobile and furniture painters, and smoking habits and atopy were seen to have a significant effect on OA occurrence. PMID- 9860246 TI - Improvement with disodium cromoglycate of neutrophil phagocytosis and respiratory burst activity in a patient with hyperimmunoglobulin E syndrome. AB - We report a case of hyperimmunoglobulin E syndrome (HIE) complicated by neutrophil deficiency which was successfully treated with oral administration of disodium cromoglycate. A 48-year-old Japanese man with HIE developed Streptococcus pneumoniae meningitis. Laboratory tests after the meningitis revealed persistent neutropenia (300-800/mm3) and defects of phagocytosis and bacterial killing by neutrophils. Administration of disodium cromoglycate was started, and neutrophil counts gradually increased to 1200-1600/mm3. The impaired neutrophil activities returned to normal. The patient improved clinically; during the 2-year treatment, he had only two brief episodes of the common cold. Disodium cromoglycate may have potential clinical use in the treatment of cases of HIE even with neutrophil deficiency. PMID- 9860247 TI - Acute pancreatitis caused by allergy to kiwi fruit. PMID- 9860248 TI - Latex allergy in clinical practice. PMID- 9860249 TI - Pizza, an unsuspected source of soybean allergen exposure. PMID- 9860250 TI - Allergy to latex condoms. PMID- 9860251 TI - Anaphylaxis by pyridostigmine. PMID- 9860252 TI - Colocalization of tight junction proteins, occludin and ZO-1, and glucose transporter GLUT1 in cells of the blood-ocular barrier in the mouse eye. AB - The facilitative glucose transporter GLUT1 is abundant in cells of the blood ocular barrier and serves as a glucose transport mechanism in the barrier. To see the relationship between the glucose transfer function and junctional proteins in the barrier, we examined the localization of GLUT1 and the tight junction proteins, occludin and ZO-1, in the mouse eye. Their localization in the retina, ciliary body, and iris was visualized by double-immunofluorescence microscopy and immunogold electron microscopy. Occludin and ZO-1 were colocalized at tight junctions of the cells of the barrier: retinal pigment epithelial cells, non pigmented epithelial cells of the ciliary body, and endothelial cells of GLUT1 positive blood vessels. Occludin was restricted to these cells of the barrier. ZO 1 was found, in addition, in sites not functioning as a barrier: the outer limiting membrane in the retina, in the cell border between pigmented and non pigmented epithelial cells in the ciliary body, and GLUT1-negative blood vessels. These observations show that localization of occludin is restricted to tight junctions of cells of the barrier, whereas ZO-1 is more widely distributed. PMID- 9860253 TI - Plasma membrane phospholipid asymmetry precedes DNA fragmentation in different apoptotic cell models. AB - Biochemical alterations occurring in many cell types during apoptosis include the loss of plasma membrane phospholipid asymmetry and nuclear DNA fragmentation. Annexin V staining detects phosphatidylserine translocation into the outer plasma membrane layer occurring during cell death, while the in situ tailing (IST or TUNEL) reaction labels the DNA strand breaks typical of apoptosis. To compare the time course of these processes we investigated methylprednisolone-induced apoptosis of rat thymocytes, topoisomerase inhibitor-induced apoptosis in the human histiocytic lymphoma cell line U937, and serum deprivation-induced apoptosis in the rat pheochromocytoma cell line, PC12. At all time points, FACS analysis and quantitative fluorescence light microscopy showed a higher proportion of annexin V-positive than IST-positive cells, with significantly different time courses in the apoptotic cell models investigated (Anova test). Results were confirmed by confocal microscopy. Our data indicate that the exposure of phosphatidylserine, a potential phagocyte recognition signal on the cell surface of apoptotic cells in vivo, precedes DNA strand breaks during apoptosis in different cell types. PMID- 9860254 TI - Differentiating cells of murine stratified squamous epithelia constitutively express plasminogen activator inhibitor type 2 (PAI-2). AB - In stratified squamous epithelia a critical balance among cell proliferation, differentiation, and death must be maintained in order for these tissues to fulfill their barrier function. Previous studies have demonstrated that plasminogen activator inhibitor 2 (PAI-2) is a product of differentiating epidermal keratinocytes, suggesting a role for this inhibitor during squamous differentiation. Furthermore, in certain tumor cell lines, overexpression of PAI 2 confers resistance to the induction of programmed cell death, suggesting cytoprotective function(s). In the present study we demonstrate that PAI-2 mRNA and protein are constitutively and uniquely expressed in differentiating cells of murine stratified squamous epithelia, including epidermis, esophagus, vagina, oral mucosa, and tongue. PAI-2 immunohistochemical localization patterns suggest a predominantly cytosolic distribution, consistent with biochemical identification of the major PAI-2 species as a 43-kDa, presumably non glycosylated protein. Functional analysis shows that the majority of epithelial PAI-2 is active. In contrast to the high levels of PAI-2 expression in stratified squamous epithelia, little or no PAI-2 is detectable in simple epithelia. These findings suggest that epithelial PAI-2 may mediate inhibition of intracellular proteinases associated with events during terminal differentiation and death that are unique to stratified squamous epithelia. PMID- 9860255 TI - Improved mRNA in situ hybridization on formaldehyde-fixed and paraffin-embedded tissue using signal amplification with different haptenized tyramides. AB - We report an optimized in situ hybridization (ISH) protocol with a rapid signal amplification procedure based on catalyzed reporter deposition (CARD) to increase the sensitivity of non-isotopic mRNA ISH on formaldehyde-fixed and paraffin embedded tissue. The CARD method is based on the deposition of haptenized tyramide molecules in the vicinity of hybridized probes catalyzed by horseradish peroxidase. Commercially available and newly synthesized haptenized tyramides, including digoxigenin-, biotin-, di- and trinitrophenyl- as well as fluorescein tyramide, were compared. The haptenized tyramides were visualized using peroxidase conjugated anti-hapten antibodies followed by the diaminobenzidine reaction. As a test system, we applied digoxigenin-labeled oligonucleotides to detect insulin and vasoactive intestinal polypeptide mRNA in pancreatic endocrine tumors and liver metastases. Our results indicate that specificity, sensitivity, and applicability of oligonucleotide mRNA ISH can be significantly improved by using chemically digoxigenin-labeled oligonucleotide probes and signal amplification by CARD. Furthermore, all tested tyramides provided approximately equal amplification efficiency. In conclusion, CARD signal amplification should further promote mRNA ISH studies on paraffin-embedded tissues and allow for multiple-target nucleic acid detection in situ. PMID- 9860256 TI - Immunohistochemical localization of S100A1 and S100A6 in postnatally developing salivary glands of rats. AB - S100 proteins are calcium-binding proteins of the EF-hand superfamily and are involved in the regulation of a number of cellular processes. The present study deals with the immunohistochemical expression of S100A1 and S10100A6 in the rat submandibular and sublingual glands during postnatal development from day 0 to 12 weeks. Expression of S100A1 was particularly concentrated in pillar and transition cells in the granular convoluted tubule (GCT) and in striated duct cells of the submandibular gland age 4 weeks to maturity. None or only weak staining for S100A1 was observed in the duct segment at 0-5 days. On the contrary, immunostaining of S100A6 was present in proacinar cells in undifferentiated submandibular gland at age 3 days to 2 weeks. S100A6 immunoreactivity in rat submandibular gland coexisted with chromogranin reactivity. It is possible that S100A6 regulates secretion of chromogranin in proacinar cells. Secretion of growth factors and biologically active peptides in the GCT are regulated by calcium signals, and S100A1 may be involved in the secretory mechanism of granular cells. S100A1 and S100A6 are potentially of great importance in secretory functions of granular cells and proacinar cells, as well as in rat salivary glands. PMID- 9860257 TI - Ki-67 (MIB 5) immunostaining of mouse lung tumors induced by 4-nitroquinoline 1 oxide. AB - We immunostained mouse lung tumors using a mouse monoclonal antibody against recombinant Ki-67 antigen (clone; MIB 5) to establish an MIB 5 immunostaining method and to determine the extent of MIB 5 labeling to monitor cell proliferation activity in mouse lung tumors. A/J mice, treated with 4 nitroquinoline 1-oxide, were killed after 18 months. One hour before killing, bromodeoxyuridine (BrdU) was injected intraperitoneally. Lung tissues including tumors were fixed with phosphate-buffered 4% paraformaldehyde and embedded in paraffin. For MIB 5 immunostaining, two antigen-retrieval buffers, citrate buffer pH 6 and TRIS-HCl buffer pH 9.5 containing 5% urea, were tested, and constant and reproducible staining was obtained only with the TRIS-HCI buffer. The mean values of the MIB 5-positive cell index (PCI), the BrdU labeling index (LI), and the mitotic cell count for adenocarcinomas were 4.6%, 2.3%, and 7/mm2, and those for adenomas were 1.2%, 0.7%, and 1.3/mm2, respectively. Each of these values was significantly higher for adenocarcinomas than for adenomas. A close correlation was seen between the MIB 5 PCI and the BrdU LI for adenocarcinomas and adenomas and between the MIB 5 PCI and the mitotic cell count in adenocarcinomas. Thus, MIB 5 immunostaining is a useful method for assessing the proliferative activity of mouse tumor tissues. PMID- 9860258 TI - CNS glia are targets for GDNF and neurturin. AB - Glial cell line-derived neurotrophic factor (GDNF) and neurturin (NTN) are two closely related growth factors reported to selectively act on distinct neuronal populations in the CNS. Both GDNF and NTN signal through a receptor complex consisting of the signal transducing subunit, Ret, and a ligand-specific binding subunit, termed GDNF family receptor (GFR)alpha-1 and GFRalpha-2, respectively. By using RT-PCR, we observed that mRNAs encoding the subunits of both receptor complexes are widely expressed throughout the developing brain, suggesting the presence of targets for these growth factors other than the ones known today. We provide evidence that these targets include glial cells. PMID- 9860259 TI - Unconventional antigen retrieval for carbohydrate and protein antigens. AB - Aldehyde fixation of tissues often adversely affects the reactivity of cellular proteins with antibodies. A most commonly used retrieval technique in immunohistochemistry is high-temperature microwave heating of sections from formaldehyde-fixed and paraffin-embedded tissues. Here we report that pretreatment of paraffin and ultrathin cryosections with N-glycanase F to remove N-glycosidically linked oligosaccharides can result in a dramatic increase in specificity and intensity of immunogold labeling for sugar moieties present on O glycosidically linked oligosaccharides. This is demonstrated in the immunolocalization of poly alpha2,8 KDN (KDN, 2-keto-3-deoxy-D-glycero-D-galacto nononic acid) of megalin in rat kidney. The mechanism of this retrieval procedure is most probably based on the elimination of sterical hindrance by large N glycosidically linked oligosaccharides. Furthermore, we demonstrate that exposure of ultrathin cryosections to acidic conditions (pH 5.5) at ambient temperature prior to immunogold labeling can result in an increased labeling intensity. This effect was observed for megalin immunoreactive sites in proximal tubular epithelia of rat kidney. It is proposed that the mechanism of this retrieval procedure is based on the depolymerization of methylen and polymethylen bridges introduced by formaldehyde in the acidic milieu. PMID- 9860260 TI - Expression of type I and II receptors for transforming growth factor beta in the adult rat testis. AB - After having established the specificity of the antibodies for the rat testis by western blot analysis, the potential target cells for transforming growth factors (TGFbetas) were identified by immunohistochemical detection of both type I (TbetaRI) and type II (TbetaRII) transducing receptors for TGFbetas in the adult rat testis in situ. Leydig cells showed a strong TbetaRII immunoreactivity whereas the TbetaRI staining was weak. Only TbetaRII was detectable in Sertoli cells. In germ cells, staining for TbetaRI was stronger than for TbetaRII and the expression of both receptors depended on the seminiferous cycle stage. TbetaRI first appeared in pachytene spermatocytes and was absent in elongated spermatids from stage XIV onwards. Labelling for TbetaRII was observed as early as the spermatogonia stage; it increased in pachytene spermatocytes at the onset of TbetaRI and disappeared in elongating spermatids from stage XI onwards. These results show that TGFbetas can affect somatic cells functions and suggest that these factors are involved in the control of meiosis and early spermiogenesis, exerting a direct effect on germ cells. PMID- 9860261 TI - Movement-related skin strain associated with goal-oriented lip actions. AB - It is generally accepted that sensory input contributes to the generation of natural movements. In most motor systems, muscle spindles, tendon organs, joint receptors, and cutaneous mechanoreceptors may provide proprioceptive information. However, the perioral area of the human face lacks muscle spindles, tendon organs, and joint receptors and is therefore a model system for the study of cutaneous afferent contributions to proprioception. This investigation examined a series of skin strains associated with lower-lip movements in human subjects to determine if such strains, which serve as stimuli for cutaneous mechanoreceptors, may underlie proprioception in the face. The results suggested that strains associated with lower-lip movements were of sufficient magnitude to elicit cutaneous mechanoreceptor discharge, as shown in recent human microneurographic studies. Further, the magnitude of multiple strains was predictive of lower-lip movement endpoints. These results highlight the potential importance of cutaneous mechanoreceptors as putative proprioceptors. PMID- 9860262 TI - High acceleration impulsive rotations reveal severe long-term deficits of the horizontal vestibulo-ocular reflex in the guinea pig. AB - While there is agreement that unilateral vestibular deafferentation (UVD) invariably produces an immediate severe horizontal vestibulo-ocular reflex (HVOR) deficit, there is disagreement about whether or not this deficit recovers and, if so, whether it recovers fully or only partly. We suspected that this disagreement might mainly be due to experimental factors, such as the species studied, the means chosen to carry out the UVD, or the nature of the test stimulus used. Our aim was to sort out some of these factors. To do this, we studied the HVOR of alert guinea pigs in response to low and high acceleration sinusoidal and high acceleration impulses after UVD by either labyrinthectomy or by vestibular neurectomy. The HVOR in response to high acceleration impulsive yaw rotations was measured before, and at various times after, either unilateral labyrinthectomy or superior vestibular neurectomy. Following UVD, there was a severe impairment of the HVOR for ipsilesional rotations and a slight impairment for contralesional rotations, after either operation. This asymmetrical HVOR deficit in the guinea pig parallels the deficit observed in humans. Between the first measurement, which was made 1 week after UVD, and the last, which was made 3 months after UVD, there was no change in the HVOR. This lack of recovery was the same after labyrinthectomy as after vestibular neurectomy. The HVOR to low and high acceleration sinusoidal yaw rotations were measured after UVD, and the results were compared with those in response to impulsive rotations. For low acceleration sinusoidal rotations (250 degrees/s2), the gain was symmetrical, although reduced bilaterally. As the peak head acceleration increased, the HVOR became increasingly asymmetric. The HVOR asymmetry for sinusoidal rotations was significantly less than for impulsive rotations that had the same high peak head acceleration (2500 degrees/s2). Our results show that the HVOR deficit after UVD is the same in guinea pigs as in humans; that it is the same after vestibular neurectomy as after labyrinthectomy; that it is lasting and severe in response to high acceleration rotations; and, that it is more obvious in response to impulses than to sinusoids. PMID- 9860263 TI - Changes in the force-sharing pattern induced by modifications of visual feedback during force production by a set of fingers. AB - We investigated force-sharing among three fingers which acted in parallel and produced ramp profiles of total force from zero to the maximal voluntary force. The feedback to the subject was provided by a visual signal on the monitor and could correspond to the sum of forces of all the fingers or to the sum of forces of two fingers, while the force of the third finger was added with a coefficient 2 or 0.5. If the subjects did not know about the distorted feedback, they showed a template-sharing pattern within the whole range of total force values. This pattern did not depend on which finger force was multiplied and by which coefficient. If the subjects knew in advance how the feedback signal would be calculated, they tried to perform the task using either only the finger whose force was multiplied by 2 or two fingers when the force of the third one was multiplied by 0.5. Further into the trial, however, they were unable to track the ramp pattern using only one or two fingers and demonstrated a search activity that could continue until the end of the trial or lead eventually to a three finger sharing pattern similar to the template pattern used in cases of undistorted feedback. We conclude that the limited number of preferred sharing pattern within the studied task reflects an organization of the fingers into a structural unit (involving one, two, or all three fingers) by the central nervous system. The availability of structural units defines the presence of stable solutions available for the system. PMID- 9860264 TI - Glutathione protects PC12 cells from ascorbate- and dopamine-induced apoptosis. AB - The role of reduced glutathione (GSH) on ascorbate- and dopamine-induced apoptosis in PC12 cells was investigated. Ascorbate is a potent reducing agent and is thus expected to protect against dopamine-induced apoptosis. However, we found that both ascorbate and dopamine killed PC12 cells and ascorbate enhanced dopamine-induced toxicity. The EC50 of cell toxicity induced by ascorbate, dopamine and dopamine plus 0.1 mM ascorbate during 24-h treatment were 0.93+/ 0.15 mM, 0.18+/-0.05 mM and 0.13+/-0.04 mM, respectively. When the medium contained 10 mM GSH, the EC50 increased approximately three- and sevenfold for ascorbate and dopamine, respectively. With increased treatment duration, no further toxic effects of ascorbate or dopamine were observed. The GSH synthesis inhibitor, DL-buthionine-(S,R)-sulfoximine (BSO), induced cell toxicity and potentiated the toxic effects of ascorbate and dopamine, suggesting that endogenous GSH participates in protecting against basal oxidative stress. We conclude that both ascorbate and dopamine induce apoptosis in PC12 cells and further that GSH protects them from apoptosis. This study indicates that the toxic effects of ascorbate are potentially due to an oxidative mechanism, similar to that induced by dopamine. PMID- 9860265 TI - Simple and choice reaction-time performance following occlusion of the anterior cerebral arteries in the rat. AB - Focal cerebral ischemia in the rat has traditionally been studied by examining the consequences of middle-cerebral artery occlusion. However, the anteriorcerebral arteries of the rat may now also be bilaterally occluded by stereotaxic injection of the vasoconstrictor endothelin-1, resulting in ischemic damage to medial prefrontal cortex and the anteromedial basal forebrain. The behavioural consequences of anterior-cerebral artery occlusion (ACAo) were studied in two experiments using simple and choice reaction-time tasks designed to dissociate response impairments from dysfunction of motivation and attention, respectively. Following ACAo, reaction-time increased post-surgery in the choice, but not simple reaction-time task. There was also an increase in incorrect choices in the choice reaction-time task. However, the impairments were independent of motivational or attentional function, which remained intact. Although the ACAo-induced ischemic damage did not disrupt motivation or attention, the results suggest that the lesion results in an executive impairment in selecting and initiating responses. PMID- 9860266 TI - Column spacing in normal and visually deprived monkeys. AB - A recent model for the development of the pattern of eye-dominance domains in primary visual cortex predicts that stimulus conditions during early visual life determine the spacing (or periodicity) of ocular dominance columns (ODC). The model predicts that normal binocular visual experience consists of highly correlated binocular stimulation and leads to relatively narrow ODC spacing, while abnormal binocular visual stimulation attendant with strabismus consists of non-correlated, incoherent, and asynchronous stimulation and leads to wider than normal ODC spacing. Evidence in support of the model has been presented for strabismus in the kitten. We tested the predictions of the model in normal monkeys and others subjected to various forms of abnormal visual experience during infancy. We identified and measured the inter-column spacing (or periodicity) in the V1 cortex of 19 adult monkeys (M. mulatta) using the cytochrome-oxidase (CO) histochemical method. There were no significant differences in the V1 inter-column spacing between normal adult monkeys (n=5) and other adult monkeys having had monocular-form deprivation (n=5), experimental anisometropia (n=5), or experimental strabismus (n=4) early in life. The quality of early binocular visual experience is not a significant determinant of the inter-column spacing in primate V1 cortex. Therefore, the model predicting an increase in the ODC periodicity with strabismus is not supported. PMID- 9860267 TI - Attentional coding for three-dimensional objects and two-dimensional shapes. Differential interference effects. AB - The role played by the attentional mechanisms that enable dominance of relevant objects over distractor objects was investigated by measuring changes in the kinematics of the reach-to-grasp movement. Subjects reached towards three dimensional (3D) stimuli while attention was diverted towards distracting information consisting of either two-dimensional (2D) projected shapes or 3D objects. Movement kinematics were influenced to a greater degree when a secondary task was performed involving a 3D object rather than a 2D projected shape. When the distractor was 3D, both the reaching and the grasping components were altered but, when it was 2D only, the reaching component was modified. It is suggested that, when attention is directed towards a distractor, it is associated with interference in the kinematics of the action towards the target. Further, the nature and dimensions of the distractor selectively influence the reach or the grasp component of a prehension movement. PMID- 9860268 TI - Altered responses to potassium in cerebellar neurons from weaver heterozygote mice. AB - The pleiotropic weaver disease is caused by the mutation of a single amino acid in the G-protein-linked inwardly rectifying K+ channel, GIRK2. In homozygous (wv/wv) animals, the disease is characterized by loss of cerebellar and dopaminergic mesencephalic neurons as well as testicular cells, which produce ataxia, fine tremors, and sterility, respectively. Heterozygous (wv/+) animals show no obvious motor impairments, although some loss of both cerebellar and dopaminergic neurons is observed and wv/+ males become sterile at 3.5 months of age. Abnormal influxes of Na+ and Ca2+ have been linked to cerebellar cell death in wv/wv animals, but it's not clear whether similar changes are observed in wv/+ animals. To discover whether changes in K+-channel function or intracellular Ca2+ concentrations ([Ca2+]i) play a role in the augmented cell loss observed in wv/+ animals when compared with +/+ animals, we studied cultured cerebellar granule cells prepared from either wv/+ or +/+ animals. Resting [Ca2+]i was elevated in wv/+ relative to +/+ animals. Further, depolarizations of cells with elevated K+ solutions elicited much smaller changes in [Ca2+]i in wv/+ animals than in +/+ animals, presumably due to altered GIRK2 channel function. Both wv/+ and +/+ cells showed similar changes in [Ca2+]i when cells were depolarized by glutamate (1 mM), suggesting that both glutamate receptors and Ca2+ channels were unchanged in wv/ + animals. In summary, our results suggest that wv/+ cerebellar granule cells exhibit elevated resting [Ca2+]i levels and altered K+-channel function, which may contribute to the developmental abnormalities and increased cell death observed. PMID- 9860269 TI - Effect of the mono- and tetra-sialogangliosides, GM1 and GQ1b, on long-term potentiation in the CA1 hippocampal neurons of the guinea pig. AB - Effects of the mono- and tetra-sialogangliosides, GM1 and GQ1b, on long-term potentiation (LTP) were investigated in the CA1 neurons of guinea-pig hippocampal slices. The magnitude of LTP induced by a strong tetanus (100 Hz, 100 pulses) was not significantly affected by application of either ganglioside. In contrast, when LTP was induced by a weak tetanus (100 Hz, 4 pulses), a significantly greater LTP was induced in the presence of either ganglioside. Similarly, when slices were incubated in low-Ca2+ (1.0-1.1 mM) medium for more than 2 h, the LTP was usually small or absent, but showed a significant increase in amplitude of population spike (A-PS) when the slices were incubated with either GM1 or GQ1b (4 5 microg/ml). In addition, the application of GQ1b (4 microg/ml) reversed the blocking effect of an NMDA-receptor antagonist, APP-5 (10 microM), on the induction of LTP and resulted in forming LTP. Based on these findings, we conclude that GM1 and GQ1b exert positive modulatory effects on the induction of LTP in hippocampal CA1 neurons and suggest that GM1 and GQ1b may participate in the induction of LTP as donors of Ca2+ ions. PMID- 9860270 TI - Representation of the midline trunk, bilateral arms, and shoulders in the monkey postcentral somatosensory cortex. AB - Single neuronal activities were recorded in the arm/trunk region of the postcentral gyrus in awake Japanese monkeys. A total of 1608 units were isolated from four hemispheres of two animals, and receptive fields (RFs) and submodalities were identified in 1162 units. Deep or skin submodality neurons were dominant in area 3a or area 3b, respectively. The deep/skin ratio increased as the recording site moved from area 3b to the more caudal areas. In areas 3a and 3b, neuronal RFs were almost exclusively on either the arm or trunk. In areas 2 and 5, neurons with RFs on the trunk decreased and those with RFs on the hand or covering more than one body part, etc. increased. We found a total of 107 neurons with bilateral RFs and 56 with ipsilateral RFs, while the rest (n=999) were with contralateral RFs. Bilateral or ipsilateral neurons of skin submodality (n=37) were found in areas 1, 2, and 5. Twenty six (70%) had RFs on the trunk and/or occiput, five on the forelimb, and the rest (n=6) on both the trunk and forelimb (the combined type). Among 33 skin bilateral neurons, 90% (n=30) had RFs across the midline. Bilateral or ipsilateral neurons responding to joint manipulation (n=104) were found in areas 2 and 5. Most of them were activated by manipulation of the shoulder and/or elbow (the proximal type, n=72, 69%). There were 25 neurons of the combined type (both the proximal and distal joints were effective, 24%). Bilateral or ipsilateral neurons of deep-others submodality (n=20) were found in areas 1, 2, and 5. The forelimb type (n=12, 60%) was dominant in this category. The combined-type neurons in both the skin- and joint manipulation categories were found only or mostly in area 5. These results indicate the presence of hierarchical processing for bilateral as well as contralateral information within the arm/trunk region of the postcentral gyrus. PMID- 9860271 TI - Efferent synaptic connections of dopaminergic neurons grafted into the caudate nucleus of experimentally induced parkinsonian monkeys are different from those of control animals. AB - This study investigated the question of whether grafted dopamine cells in the striatum of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated monkeys form synapses and, if they do, whether their postsynaptic targets were the same as those in control monkeys or in previous studies in rats. Electron-microscopic single immunostaining was performed for tyrosine hydroxylase on vibratome sections prepared from the head of the caudate nucleus of controls and MPTP treated African green monkeys (Cercopithecus aethiops sabaeus) that received a graft. Furthermore, correlated light- and electron-microscopic double immunostaining was carried out for tyrosine hydroxylase and calbindin in the same brain area of MPTP-treated plus grafted animals. In control monkeys, the majority (97%) of dopamine boutons terminate on spines that were also synaptic targets of immunonegative boutons forming asymmetric synaptic contacts: synaptic triads. In MPTP-treated, grafted animals, the majority of transplanted dopamine cells terminate on dendritic shafts (67%) and somata (32%), and only a few (1.33%) form axospine synapses. The results of the double immunostaining experiments indicated that these newly formed axosomatic and axodendritic synapses are associated with calbindin-immunoreactive, medium-sized, spiny striatonigral projection neurons. These observations indicate that: (1) dopamine from transplanted embryonic tissue acts via synaptic contacts on host neurons; (2) the primary synaptic targets of transplanted dopamine cells are not spines but dendrites and somata of host neurons; (3) these target neurons are the same as in control animals; and (4) comparing these observations with results of control and grafted rats, there are major species differences between rats and monkeys in the dopamine innervation of both control and transplanted animals. PMID- 9860273 TI - Microsaccades differentially modulate neural activity in the striate and extrastriate visual cortex. AB - Saccadic eye movements in primates continually shift the location at which a given stimulus strikes the retina. Even during periods of steady fixation, microsaccades frequently jerk the center of gaze by small but resolvable distances, yet perception remains stable and continuous, uninterrupted by sudden jumps or shifts. The effect of such fixational eye movements on the activity of single neurons was examined in several regions of the visual cortex in macaque monkeys. We found that the firing of many neurons in striate and extrastriate cortex is profoundly influenced by saccades much smaller than the neurons' receptive fields. In striate cortex (V1) many cells showed a transient decrease in their firing shortly following a saccade. In sharp contrast, cells in the extrastriate areas V2 and V4 showed strong excitatory responses that closely coincided in time with the striate depression. No appreciable activity change was observed in the inferotemporal cortex (IT) following saccades. This activity pattern is consistent with the notion that topographic extrastriate areas receive extraretinal input associated with saccadic events. Such signals may be necessary for the stable perception of objects and scenes during eye movements, mediating the mapping between central object representations and the constantly changing retinotopic input. PMID- 9860272 TI - Co-localization of corticotropin-releasing hormone with glutamate decarboxylase and calcium-binding proteins in infant rat neocortical interneurons. AB - Corticotropin releasing hormone (CRH) has been localized to interneurons of the mammalian cerebral cortex, but these neurons have not been fully characterized. The present study determined the extent of co-localization of CRH with glutamate decarboxylase (GAD) and calcium-binding proteins in the infant rat neocortex using immunocytochemistry. CRH-immunoreactive (ir) neurons were classified into two major groups. The first group was larger and consisted of densely CRH immunostained small bipolar cells, predominantly localized to layers II and III. The second group of CRH-ir cells was lightly labeled and included multipolar neurons mainly found in deep cortical layers. Co-localization studies indicated that the vast majority of CRH-ir neurons, including both bipolar and multipolar types, was co-immunolabeled for GAD-65 and GAD-67. Most multipolar, but only some bipolar, CRH-ir neurons also contained parvalbumin, while CRH-ir neurons rarely contained calbindin or calretinin. These results indicate that virtually all CRH ir neurons in the rat cerebral cortex are GABAergic. Furthermore, since parvalbumin is expressed by cortical basket and chandelier cells, the co localization of CRH and parvalbumin suggests that some cortical CRH-ir neurons may belong to these two cell types. PMID- 9860274 TI - Coordination among the body segments during reach-to-grasp action involving the trunk. AB - To understand the internal representations used by the nervous system to coordinate multijoint movements, we examined the coordination among the body segments during reach-to-grasp movements which involve grasping by the hand and reaching by the arm and trunk. Subjects were asked to reach and grasp an object using the arm only, the trunk only, and some combinations of both arm and trunk. Results showed that kinematic parameters related to the transport component of the arm and the trunk, such as peak velocity and time to peak velocity, varied across conditions and that the coordination pattern between the arm and trunk was different across conditions. However, parameters related to the grasp component, such as peak aperture, time to peak aperture, and closing distance, were invariant, regardless of whether the hand was delivered to the target by the arm only, the trunk only, or both. We hypothesize that a hierarchy of motor control processes exists, in which the reach and grasp components are governed by independent neuromotor synergies, which in turn are coordinated temporally and spatially by a higher-level synergy. PMID- 9860275 TI - Distribution of adenosine A1 receptors in primary visual cortex of developing and adult monkeys. AB - Adenosine is an endogenous neuromodulator, whose actions in the brain are mediated by several, highly specific receptor systems. This study examined the distribution of adenosine A1 receptors in the primary visual cortex of developing and adult monkeys by way of radioligand-binding autoradiography. Receptor density was found to be highest in layers 2/3 and 5 of adult striate cortex and lowest in layer 4. A similar pattern was found in infant monkeys that were in the critical period of development. Furthermore, removal of visual stimulation appeared to have no effect on receptor density in any of the layers. These results show that levels of adenosine A1 receptors are not affected within ocular dominance columns after removal of visual input and that the geniculorecipient layer is less likely to be influenced by endogenous adenosine than other layers during early development and in the adult. PMID- 9860276 TI - Independent coactivation of shoulder and elbow muscles. AB - The aim of this study was to examine the possibility of independent muscle coactivation at the shoulder and elbow. Subjects performed rapid point-to-point movements in a horizontal plane from different initial limb configurations to a single target. EMG activity was measured from flexor and extensor muscles acting at the shoulder (pectoralis clavicular head and posterior deltoid) and elbow (biceps long head and triceps lateral head) and flexor and extensor muscles acting at both joints (biceps short head and triceps long head). Muscle coactivation was assessed by measuring tonic levels of electromyographic (EMG) activity after limb position stabilized following the end of the movements. It was observed that tonic EMG levels following movements to the same target varied as a function of the amplitude of shoulder and elbow motion. Moreover, for the movements tested here, the coactivation of shoulder and elbow muscles was found to be independent - tonic EMG activity of shoulder muscles increased in proportion to shoulder movement, but was unrelated to elbow motion, whereas elbow and double-joint muscle coactivation varied with the amplitude of elbow movement and were not correlated with shoulder motion. In addition, tonic EMG levels were higher for movements in which the shoulder and elbow rotated in the same direction than for those in which the joints rotated in opposite directions. In this respect, muscle coactivation may reflect a simple strategy to compensate for forces introduced by multijoint limb dynamics. PMID- 9860277 TI - The small-cell variant of mycosis fungoides. A clinicopathological and quantitative electron microscopic study on 14 patients. AB - Small-cell variants of Sezary syndrome and mycosis fungoides (MF) have been described. However, in these studies the nuclear area of the small-cell variant of MF (SC-MF) as compared to histological classical MF (CL-MF) was not characterized objectively by quantitative electron microscopy. In a 14-year follow-up period, of a total of 76 patch/plaque stage MF patients seen in the Department of Dermatology of the University Hospital Utrecht, 14 (18%) had an infiltrate composed of atypical lymphocytes characterized by a distinctly smaller cell diameter and smaller, hyperchromatic, deeply indented nuclei as compared to the usual cell type of MF. The aim of the investigation was to confirm this observation objectively using quantitative electron microscopy (morphometry) and to define SC-MF as compared to CL-MF. The study was performed on the 14 patients with SC-MF, and 10 patients with clinical and histological CL-MF and 4 patients with chronic eczema. Electron micrographs of sections obtained from each biopsy were analysed by computer to produce the following data: a nuclear contour index (NCI), the mean nuclear area (MNA), the mean nuclear area of the cells above the 75th percentile (P75NA) and the percentage of cells larger than 30 microm2. The values of MNA differed significantly between patients with SC-MF and those with CL-MF (17.6 vs 23.2 microm2; P = 0.02), as did the values of P75NA (20.7 vs 27.9 microm2; P = 0.01). The NCI of the SC-MF and CL-MF patients were similar. These results are consistent with our observations that SC-MF does indeed exist. PMID- 9860279 TI - Expression of CD30 on T helper cells in the inflammatory infiltrate of acute atopic dermatitis but not of allergic contact dermatitis. AB - The CD30 molecule has been proposed as a marker for a subset of CD4+CD45RO+ (memory) T cells with potent B cell helper activity producing IL-5 and IFN-gamma and as a specific marker for Th2 cells. Recently, an association has been demonstrated between elevated serum levels of soluble CD30, which is shed by CD30+ cells in vitro and in vivo, and atopic dermatitis but not respiratory atopic disorders or allergic contact dermatitis. We studied the expression of CD30 in the inflammatory infiltrate of atopic dermatitis compared with that of allergic contact dermatitis, with special regard to skin disease activity (acute vs subacute/ chronic). Biopsies were obtained from 16 patients suffering from atopic dermatitis (acute n = 6, subacute/ chronic n = 10), from 7 patients with acute allergic contact dermatitis and from 5 positive patch-test reactions. Paraffin-embedded as well as snap-frozen material was stained with anti-CD30 and anti-CD45RO mAbs according to standard procedures. Double-staining procedures for CD30CD3, CD30CD4, CD30CD45RO and CD30CD68 were also performed. Abundant CD45RO+ cells were detected both in atopic dermatitis and in allergic contact dermatitis lesions. We found scattered CD30+ cells in only one of six formalin-fixed paraffin-embedded acute atopic dermatitis biopsies, but in all of the respective snap-frozen specimens, possibly because CD30 expression on atopic dermatitis infiltrating cells is weak and sensitive to formalin fixation and paraffin embedding. CD30CD3 and CD30CD4 double staining identified CD30+ cells to be helper T lymphocytes. No significant CD30 expression (either in paraffin-embedded or in frozen material) could be found in subacute/chronic atopic dermatitis lesions or in any of the specimens of allergic contact dermatitis. The results suggest a specific regulatory function of CD30+ T cells in acute atopic dermatitis. With respect to the view that CD30 is a marker for Th2 cells, our observations confirm previous findings that Th2 cells predominate in the infiltrate particularly of acute atopic dermatitis. CD30 expression in acute atopic dermatitis but not in acute allergic contact dermatitis might be helpful in the histological differentiation of these disorders and in the further characterization of atopy patch testing. PMID- 9860278 TI - Abnormalities of basal cell keratin in epidermolysis bullosa simplex do not affect the expression patterns of suprabasal keratins and cornified cell envelope proteins. AB - Basal keratins, suprabasal keratins, filaggrin, and cornified cell envelope (CCE) precursor proteins are expressed during the differentiation of epidermal keratinocytes. These molecules are coordinately expressed during epidermal differentiation. The present study investigated the expression patterns of keratins and CCE precursor proteins in 15 patients with epidermolysis bullosa simplex (EBS), which is caused by mutations in the genes that encode for the basal keratins, keratins 5 and 14. The patterns of expression of keratins 5, 14, 1 and 10, filaggrin, and of the three major CCE precursor proteins, involucrin, loricrin and small proline-rich proteins 1 and 2 (SPRs), were studied immunohistochemically and by electron microscopy. In 14 of the 15 patients with EBS, the distribution pattern of keratins was not altered. In one neonate with EBS, basal cell keratins were expressed in the suprabasal layers. Ultrastructurally, numerous clumped tonofilaments were observed in the basal and suprabasal cells. In all cases, findings were positive for filaggrin in the granular cells, with positivity for involucrin in the upper spinous and granular cells. The upper spinous cells and granular cells were positive for SPRs 1 and 2, and loricrin was expressed in granular cells. Ultrastructurally, no marked abnormality was observed in the suprabasal layers such as a decrease in, or agglutination of, keratin filaments, except in one neonate. A CCE about 15 nm thick was formed normally in the cell membrane of cornified cells. The patterns of distributions of basal cell keratins, suprabasal keratins, filaggrin, and CCE precursor proteins, as well as the ultrastructural findings, resembled those of normal skin. Thus, the abnormality in basal cell keratins in patients with EBS did not appear to alter the patterns of expression of the keratins and CCE precursor proteins. PMID- 9860280 TI - The importance of CD54 and CD86 costimulation in T cells stimulated with Candida albicans and Dermatophagoides farinae antigens in patients with atopic dermatitis. AB - A number of studies have demonstrated an increased frequency of allergen-specific T cells producing increased amounts of interleukin-4 (IL-4) and IL-5, but little interferon-y in both the peripheral blood and skin lesions of patients with atopic dermatitis (AD). In this study, to further clarify the characteristics of T cells obtained from AD patients, we examined the dependency of the antigen specific proliferation of peripheral blood mononuclear cells (PBMC) from AD patients on costimulatory molecules. The antigens used were Candida albicans and Dermatophagoides farinae, for which AD patients show increased levels of IgE antibodies. PBMC from control healthy donors stimulated with these antigens incorporated [3H]-thymidine much more than PBMC from AD patients. The addition of anti-CD54, -CD40, -CD80 and -CD86 monoclonal antibodies to the cultures showed that the PBMC required only CD54 and CD86 for stimulation with C. albicans, but required CD54, CD80 and CD86 for stimulation with D. farinae. Among these monoclonal antibodies, the anti-CD54 antibody suppressed the proliferative responses of most PBMC, most effectively followed by the anti-CD86 antibody. However, there were no significant differences in the requirement for costimulatory molecules of PBMC proliferation stimulated with C. albicans or D. farinae between AD patients and healthy donors. Since many studies have suggested that T-helper type 1 and T-helper type 2 immune responses are different in their dependency on CD80 or CD86 costimulation, our present results suggest that the allergen-specific T cells of AD patients are not completely shifted to a T-helper type 2 subset. PMID- 9860281 TI - Nerve growth factor in normal and psoriatic skin equivalent models. AB - Our objective was to determine the pattern and time course of nerve growth factor expression in an established skin equivalent model that we have used in the past to study wound healing and psoriasis phenotypes. Skin equivalents were constructed in triplicate using normal neonatal foreskin keratinocytes plated on collagen gels containing fibroblast lines. These lines were derived from five specimens of psoriatic lesions, three specimens of normal skin from patients with psoriasis, and three specimens of eyelid skin from normal donors. Immunohistochemistry and a monoclonal nerve growth factor-b antibody were used to determine the pattern of protein staining over 2 weeks. We looked at the wound healing phenotype using the skin equivalent model for 7-14 days. When keratinocytes invaginate into the dermis of skin equivalents (beginning at around 7 days of growth), dark staining of nerve growth factor was seen under the basal membrane zone, suggesting that nerve growth factor serves in the development of the basal membrane zone and the epidermis, and may influence the migration of nerves through the basal membrane zone into the regenerated skin. PMID- 9860282 TI - SLS-irritated human skin shows no correlation between degree of proliferation and TEWL increase. AB - It is well known that cutaneous irritants influence epidermal proliferation but the pathogenesis is poorly understood. Recent investigations have shown that the skin barrier integrity influences the proliferation of the basal keratinocytes. Our question was whether the proliferating activity of keratinocytes is indeed regulated by the degree of skin barrier damage or by a direct toxic action of the irritant on the keratinocytes. Therefore various degrees of skin irritation were induced by the application of 0.1%, 0.5% and 2% sodium lauryl sulphate (SLS) solution to the forearm skin of six healthy volunteers. This experiment was performed to evaluate the relationship between SLS concentration and epidermal proliferation. In a second experiment another 14 volunteers were treated with a single SLS concentration (0.5%) to look for interindividual differences in the patterns of skin reaction and susceptibility to the irritant. Skin barrier function was evaluated by measurements of transepidermal water loss (TEWL) before and after irritation. Punch biopsies were taken after 96 h from exposed areas and from unexposed normal skin. Dividing keratinocytes were identified immunocytochemically using three different monoclonal antibodies: PCNA, MIB 1 and KiS1. Exposure to SLS resulted in concentration-dependent increases in both TEWL and epidermal proliferation. However, no significant correlation could be found between the degree of hyperproliferation and the TEWL changes. The results suggest that epidermal proliferation is modulated by a direct interaction of the surfactant with the keratinocytes and/or by release of mediators rather than the consequence of a barrier disturbance. PMID- 9860283 TI - A novel in situ method for the detection of deficient transglutaminase activity in the skin. AB - Autosomal recessive congenital ichthyoses are disorders of epidermal cornification, but are clinically and etiologically heterogeneous. Some cases, known as lamellar ichthyosis, are caused by mutations in the TGM1 gene encoding transglutaminase 1, which result in markedly diminished or lost enzyme activity and/or protein. In some cases, this enzyme is present but there is little detectable activity, and in other clinically similar cases, transglutaminase 1 levels appear to be normal. Since conventional enzyme assays and mutational analyses are tedious, we developed a novel assay for the rapid screening of transglutaminase 1 activity using covalent incorporation of biotinylated substrate peptides into skin cryostat sections. Coupled with immunohistochemical assays using transglutaminase 1 antibodies, our method allows rapid identification of those cases caused by alterations in this enzyme. PMID- 9860284 TI - Effects of diclofenac on experimental streptococcal necrotizing fasciitis (NF) in rabbit. AB - Aggravation of necrotizing fasciitis (NF) by the administration of non-steroidal antiinflammatory drugs (NSAIDs) has recently been suggested. A rabbit model of streptococcal NF was used to study the effects of parenteral administration of an NSAID on NF evolution and outcome. Of 16 rabbits inoculated with a Streptococcus pyogenes suspension together with staphylococcal alpha toxin, 8 were treated with two doses of 4 mg/kg diclofenac on day 1 after inoculation. Clinical, bacteriological and histological studies were performed until day 10. Under our experimental conditions, NSAID treatment significantly limited NF extension. A specific inverse relationship between the extent of inflammation and bacterial density in NF lesions was observed on day 1 after inoculation in the treated group suggesting that the greater severity of NF in humans treated with an NSAID could be due to the therapeutic delay induced by the misleading clinical effects of the NSAID, and not to inhibition of antibacterial defence. PMID- 9860285 TI - Loss of water from the stratum corneum induces epidermal DNA synthesis in hairless mice. AB - Many clinical studies have shown that low humidity has a deleterious effect on skin, but the mechanisms involved are poorly understood. To clarify the changes that occur in skin, we examined epidermal cell proliferation in mice kept in a dry (relative humidity < 10%) or a moist (relative humidity > 90%) environment. In animals exposed to low humidity, epidermal DNA synthesis started to increase within 12 h, reaching twice the original level, and the increased level was maintained for up to 5 days. The transepidermal water loss (TEWL) of mice kept for 12 h in the dry environment was the same as that of mice kept in the moist environment, but the skin conductance was lower. The increase in epidermal DNA synthesis following exposure to the dry environment was inhibited by topical application of petrolatum. It is concluded that loss of water from the stratum corneum induces epidermal cell proliferation within 12 h, and this change occurs in the absence of apparent cutaneous barrier dysfunction. PMID- 9860286 TI - In vitro study of the failure of skin surface after influence of hydration and preconditioning. PMID- 9860287 TI - Stability and cellular studies of [rac-1,2-bis(4-fluorophenyl) ethylenediamine][cyclobutane-1,1- dicarboxylato]platinum(II), a novel, highly active carboplatin derivative. AB - The synthesis of the diastereomeric [1,2-bis(4 fluorophenyl)ethylenediamine][cyclobutane-1, 1-dicarboxylato]platinum(II) complexes, rac- and meso-4F-Pt(CBDC), the evaluation of their structures, their tumor-inhibiting properties and their stability in physiological environment are described (reference complexes: the dichloro- and sulfatoplatinum(II) analogues, carboplatin and cisplatin). The most interesting diastereomer, rac-4F-Pt(CBDC), equals cisplatin and surpasses carboplatin in its effect on human breast cancer cell lines (MCF-7 and MDA-MB-231). Rac-4F-Pt(CBDC) is largely insensitive against attack of nucleophiles e.g. Cl-, a prerequisite for sufficient stability in vivo and for fewer side effects. In accordance with this, in vitro studies on the binding of rac-4F-Pt(CBDC) to albumin, the main plasma protein, show that the free, non-protein-bound fraction is relatively high, coming close to that of carboplatin. These properties are of importance for the transferability of the promising effects found in the cell culture experiments to in vivo conditions. The distinctly better anti-breast cancer activity of rac-4F-Pt(CBDC) than of carboplatin has been attributed to its ability to accumulate in the tumor cells. The human ovarian cancer cell line NIH-OVCAR-3 is also strongly inhibited by rac 4F-Pt(CBDC). PMID- 9860288 TI - Inhibition of tumour cell invasion by protease inhibitors: correlation with the protease profile. AB - The invasive potential of eight established human tumour cell lines of different origin has been studied in the Matrigel assay. Between 25% and 70% of the cells migrated through the Matrigel layer within 24 h, indicating that invasiveness varies with the cell type. Semiquantitative measurements of the proteases MMP-2 and MMP-9, and cathepsins B and L were performed in these cell lines and the cell culture media. High invasive potential was found in those cell lines expressing high levels of cathepsins B and L or matrix metal proteases (MMP), either alone or in combination. Overexpression of one of these enzymes is enough to explain a high invasive potential of a cell line. Selective protease inhibitors at 10 nM concentration in the culture medium were used to inhibit the migration of tumour cells in the Matrigel assay. The MMP inhibitor Batimastat reduced the invasive potential of all cell lines studied independently of the MMP expression. The effect of cysteine protease inhibitors was strongly correlated with the protease profile of the tumour cell line. Our findings support the hypothesis of a very complex activation cascade of matrix-degrading proteolytic enzymes and they underline the need to analyse the protease profile of any tumour before beginning an antiproteolytic tumour treatment. PMID- 9860289 TI - Emergence of higher levels of invasive and metastatic properties in the drug resistant cancer cell lines after the repeated administration of cisplatin in tumor-bearing mice. AB - To establish a more suitable model for reflecting biological aggressiveness in clinically recurrent cancers after chemotherapy, we made the in-vitro-established cisplatin-resistant cell lines, by exposing the parental tumor cell lines to cisplatin in a culture system, and also the in-vivo-established cisplatin resistant cell lines by repeated cisplatin administration to parental tumor bearing mice. Although both cell lines similarly demonstrated a clinically relevant low level of drug resistance (from 1.5 to 2.9 times more resistance to cisplatin than their parental cell lines), only the in-vivo-established cisplatin resistant cell lines showed significantly enhanced metastatic properties with a 2.1- to 3.4-fold increase in the number of lung metastatic nodules. These enhanced metastatic properties were caused by tumor invasiveness in combination with various levels of enhancement of cell attachment, proteolytic enzyme activity and cell motility. We concluded that anticancer drugs such as cisplatin could promote tumor progression only in the drug-resistant cell lines established in vivo. As a result, these cell lines are considered to be a more faithful and useful model for expressing biological aggressiveness in clinically recurrent cancers after chemotherapy than the conventional drug-resistant cell lines established in vitro. PMID- 9860290 TI - The effect of antibody against vascular endothelial growth factor on tumor growth and metastasis. AB - Vascular endothelial growth factor (VEGF), a very important in the process of tumor angiogenesis, was chosen as a target in a study to determine whether manipulation of angiogenesis with antibody against VEGF may interrupt tumor growth and metastasis. Anti-VEGF antibody was obtained from immunized rabbits, purified on an affinity column, and identified as neutralized antibody by Mile's assay. IVTA2MA891, a murine spontaneous breast cancer with a high rate of metastasis in lung in TA2 x 615 F1 mice, was chosen as an animal model in this study, because of the high expression of VEGF in the primary tumor as well as in the lung metastatic tumor. The anti-VEGF antibody could inhibit growth of S180 sarcoma in a dose-dependent manner, and the inhibition rate could reach 41.0% with a dose of 200 microg mouse(-1) day(-1). Anti-VEGF antibody could inhibit tumor growth by 76.2% in nude mice bearing human gastric cancer (MGC 803). When anti-VEGF antibody was combined with 131I-3H11, a murine monoclonal antibody conjugated with 131I, only one of five nude mice developed tumor and 84.0% more inhibition of tumor growth was obtained in comparison with treatment by 131I-3H11 alone. The growth of the primary tumor was inhibited by 44.0% and the number and size of the metastatic foci in the lungs were reduced by 73.0% and 83.7% respectively in the animal model, with a high rate of metastasis in lung. The anti-VEGF antibody may be potentially useful for clinical treatment of cancer and metastasis. PMID- 9860291 TI - Restoration of the p16 gene is related to increased radiosensitivity of p16 deficient lung adenocarcinoma cell lines. AB - To assess the role played by p16 gene expression in the radiosensitivity of human lung cancers, we transferred exogenous p16 genes into p16-deficient H460 and A549 lung adenocarcinoma cell lines and compared the cell survival curve in vitro after irradiation. The surviving fraction of the p16-transfected A549p16 and H460p16 cells that expressed exogenous p16 mRNA or protein was lower than those of the parental and negative control cells. The rapid exit of the p16-transfected cells from the G2/M phase in the cell cycle, both before and after irradiation, possibly contributes to the increased radiosensitivity of our experimental p16 transfected lung adenocarcinoma cell lines. We conclude that exogenous p16 gene may be another important factor controlling the intrinsic cellular radiosensitivity of cancer. PMID- 9860292 TI - Bendamustine as salvage treatment in patients with advanced progressive breast cancer: a phase II study. AB - A phase II pilot study of bendamustine as salvage treatment in patients with advanced breast cancer was performed to determine the objective response rates and make further observations on the toxicity of this drug. A group of 37 patients, pretreated with chemotherapy for advanced disease, entered the trial. Treatment consisted of 150 mg/m2 bendamustine on days 1 and 2 of a 4-week treatment course. Patients continued to receive treatment until complete remission and then two further courses, until tumour progression or unacceptable toxicity ensued. A total of 36 patients received at least one treatment course and were assessable for toxicity; 33 patients were evaluable for treatment results. Dose-limiting grade 3 and 4 WHO toxicity occurred in 5 and 3 patients respectively; 27% of patients entered complete or partial tumour remission. The median time to tumour progression was 2 months with a range of 1-14 months. The efficacy of bendamustine was apparently independent of pretreatment with anthracyclines, suggesting a lack of cross-resistance between bendamustine and anthracyclines. It can be concluded that bendamustine in the dose and application schedule used here is active in the salvage therapy of women with advanced breast cancer. The toxicity was acceptable. Future studies have to confirm the data of this pilot trial and to define the role of bendamustine in the combination chemotherapy of metastatic breast cancer that has been suggested by previous trials. PMID- 9860293 TI - Changes of immunological markers after autologous peripheral blood stem cell transplantation. AB - PURPOSE: Recently high-dose chemotherapy with peripheral blood stem cell transplantation (PBSCT) has become an important treatment for hematological and solid tumors. METHODS: Immunological parameters were examined before and after PBSCT in 9 patients with lung cancer and 13 patients with malignant lymphoma. Findings were compared with those for bone marrow transplantation (BMT). Peripheral blood cells were analyzed for phenotype and the levels of cytokines and soluble factors were measured. RESULTS: After PBSCT, activated T cells (CD3+HLA-DR+ cells, CD8+HLA-DR+ cells) and suppressor/cytotoxic T cells (CD8+CD11b- cells) were significantly higher in the patients with lung cancer than in those with malignant lymphoma. Serum levels of interleukin-4 and soluble interleukin-2 receptor were also significantly higher in the patients with lung cancer than in those with lymphoma. On the other hand, the serum levels of interferon gamma, tumor necrosis factor alpha, interleukin-6, soluble human leukocyte antigen class 1, and soluble thrombomodulin were significantly increased after bone marrow transplantation. The transfused peripheral stem cells of lung cancer and lymphoma patients had a similar number of granulocyte/macrophage-colony-forming units, but lung cancer patients had significantly more CD34-positive cells. CONCLUSION: By reinfusing large numbers of autologous immune cells, PBSCT may accelerate immune reconstitution, with T cells being likely to have a marked therapeutic potential. The changes after PBSCT were greater in patients with lung cancer than in lymphoma patients. These blood cells are potent mediators of anticancer activity and could play an important role in the elimination of autologous malignant cells. PMID- 9860294 TI - Evaluation of methylation analysis for diagnostic testing in 258 referrals suspected of Prader-Willi or Angelman syndromes. AB - Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are distinct neurodevelopmental disorders with interrelated genetic mechanisms because genomic imprinting within the chromosome 15q11-13 region affects both the PWS and the AS locus. Methylation analysis is one method of distinguishing between the maternally and paternally inherited chromosome 15. Here we present clinical and molecular data on a large series of 258 referred patients, evaluated with methylation analysis: 115 with suspected PWS and 143 with suspected AS. In these patients, the clinical phenotype was graded into three groups: classical (group 1); not classical but possible (group 2); not classical and unlikely (group 3). For PWS, a fourth group consisted of hypotonic babies. DNA methylation analysis confirmed the diagnosis of PWS in 30 patients (26%) and AS in 28 patients (20%). For 21 PWS patients the mechanism was established: 15 had deletions, 4 had uniparental disomy (UPD) and 2 a presumed imprinting defect. Clinically all those with an abnormal methylation pattern had the classical phenotype and none of those with a normal methylation pattern had classical PWS. For 23 AS patients in whom a mechanism was established, 17 had a deletion, 3 had UPD and 3 had a presumed imprinting defect. There was greater clinical overlap in AS, with 26 classical AS patients having a normal methylation pattern while an abnormal methylation pattern was seen in one patient from group 2. In addition, there were a further 40 patients with a normal methylation pattern in whom AS was still a possible diagnosis. Our conclusion is that methylation analysis provides an excellent screening test for both syndromes, providing approximately 99% diagnosis for PWS and for AS, a 75% diagnostic rate, supplemented for the remaining 25% with an essential basic starting point to further investigations. PMID- 9860295 TI - Transmission disequilibrium and sequence variants at the leptin receptor gene in extremely obese German children and adolescents. AB - Genetic determinants of the degree of obesity and body fat distribution have been demonstrated by family studies. The heritability has been estimated to be in the range 0.2-0.7. Mutation leading to obesity in humans has been described for only two genes, one of them the leptin gene. The leptin gene codes for a cytokine secreted by fat cells that binds to the leptin receptor (Lep-R), which exerts some of its biological functions by expression in the brain. Hence, the Lep-R gene appears to be a promising candidate for the determination of obesity in humans. We isolated genomic DNA clones from the Lep-R gene region and identified a new polymorphic microsatellite marker (OBR-CA) within 80 kb of the translation start of Lep-R. We genotyped this and a second, intragenic microsatellite marker (D1S2852) in 130 nuclear families consisting of extremely obese children and adolescents and both parents. Using the most frequent parental allele of both markers, our analysis revealed a significant transmission disequilibrium for the 266-bp allele of D1S2852 (corrected P-value=0.042). No significant result was obtained with the most frequent allele of OBR-CA (corrected P-value=1.0). However, two rare alleles showed transmission disequilibrium and were subsequently used for constructing a haplotype with the 266-bp allele. This haplotype had a transmission rate of 80% (nominal P-value=0.02). In order to identify the underlying mutation, we sequenced all coding exons of Lep-R and the partially overlapping gene encoding the obese receptor gene-related protein (ob rgrp) in individuals carrying this haplotype. We found one new mutation (Ser675Thr) in the Lep-R gene in one proband and several other mutations known to be not associated with obesity in other study groups. As this new mutation cannot explain our positive linkage result, the transmission disequilibrium of the 266 bp allele and the high transmission rate of the identified haplotype point towards a mutation in close proximity to marker D1S2852. PMID- 9860296 TI - Confirmation of FWT1 as a Wilms' tumour susceptibility gene and phenotypic characteristics of Wilms' tumour attributable to FWT1. AB - A susceptibility gene for Wilms' tumour (WT), designated FWT1, was previously mapped to chromosome 17q12-q21 by linkage analysis of a single family. We now confirm the existence of this gene by analysis of additional cases in the original family (3-point LOD score=5.69), and by detecting strong evidence of linkage to this region in an unrelated pedigree with seven cases of WT (3-point LOD score=2.56). Analysis of 11 smaller WT families confirms that there is genetic heterogeneity in familial WT, as three families exhibit strong evidence against linkage to FWT1. One of these was subsequently found to have a predisposing WT1 mutation. However, the other two families show evidence against both FWT1 and WT1, suggesting that at least one further familial WT gene exists. Analysis of the phenotype of 16 WT cases from the families linked to FWT1 demonstrates that they present at a significantly older age and a significantly later stage than both sporadic WT and the six cases from two families unlinked to either FWT1 or WT1. The results confirm the role of FWT1 in susceptibility to WT, provide strong evidence for genetic heterogeneity in familial WT and suggest there are phenotypic differences between familial WT due to FWT1, familial WT due to other genes and non-familial WT. PMID- 9860297 TI - The nuclear-encoded human NADH:ubiquinone oxidoreductase NDUFA8 subunit: cDNA cloning, chromosomal localization, tissue distribution, and mutation detection in complex-I-deficient patients. AB - We report the cloning of the cDNA sequence of the nuclear-encoded NDUFA8 subunit of NADH: ubiquinone oxidoreductase, the first mitochondrial respiratory chain complex. The NDUFA8 open reading frame (ORF) includes 519 bp and encodes 172 amino acids (Mr=20.1 kDa). The human cDNA sequence shows 86.2% identity with the bovine sequence, whereas the human NDUFA8 amino acid sequence is 87.8% similar to its bovine PGIV protein counterpart. Both human and bovine NDUFA8 contain a conserved cysteine motif. Polymerase chain reaction analysis of rodent/human somatic cell hybrids maps the human NDUFA8 gene to chromosome 9. A multiple tissue blot has revealed the highest NDUFA8 mRNA expression in human heart, skeletal muscle, and fetal heart. Mutation analysis of the NDUFA8 fibroblast cDNA in 20 patients with an isolated enzymatic complex I deficiency in cultured skin fibroblasts has revealed two polymorphisms, one within the ORF and the other in the 3' untranslated region of the NDUFA8 cDNA sequence. The allelic frequency of both polymorphisms was similar in controls and complex-I-deficient patients. PMID- 9860298 TI - Mitotic and meiotic instability of the CAG trinucleotide repeat in spinocerebellar ataxia type 1. AB - Spinocerebellar ataxia type 1 (SCA1) is an autosomal, dominantly inherited neurodegenerative disease caused by an unstable CAG trinucleotide repeat expansion in the ataxin-1 gene located on chromosome 6p22-p23. The expanded CAG repeat is unstable during transmission, and a variation in the CAG repeat length has been found in different tissues, including sperm samples from affected males. In order further to examine the mitotic and meiotic instability of the (CAG)n stretch we have performed single sperm and low-copy genome analysis in SCA1 patients and asymptomatic carriers. A pronounced variation in the size of the expanded allele was found in sperm cells and peripheral blood leucocytes, with a higher degree of instability seen in the sperm cells, where an allele with 50 repeat units was contracted in 11.8%, further expanded in 63.5% and unchanged in 24.6% of the single sperm analysed. We found a low instability of the normal alleles; the normal alleles from the individuals carrying a CAG repeat expansion were significantly more unstable than the normal alleles from the control individuals (P<0.001), indicating an interallelic interaction between the expanded and the normal alleles. PMID- 9860299 TI - Exon 1 donor splice site mutations in the porphobilinogen deaminase gene in the non-erythroid variant form of acute intermittent porphyria. AB - Acute intermittent porphyria (AIP) is an autosomal dominant disorder caused by a partial defect of the heme biosynthesis enzyme, porphobilinogen deaminase (PBGD). PBGD is encoded by two distinct mRNA species expressed in a tissue-specific manner from a single gene. One transcript is expressed in erythroid tissues, while the housekeeping transcript is expressed in all tissues. In classical AIP (95% of cases) the housekeeping and the erythroid-specific enzymes both have half normal activity in erythroid and non-erythroid tissues, whereas in the variant non-erythroid form of the disease the enzymatic defect is present only in non erythroid cells. A large allelic heterogeneity of mutations (n>135) has been demonstrated in classical AIP, but to date only three different mutations have been characterized in the non-erythroid variant form of AIP. We describe the molecular abnormalities responsible for the non-erythroid variant form of AIP in two French and two German unrelated AIP patients with normal PBGD activity in the erythrocytes. Three different splicing defects located in the intron 1 donor splice site were identified: a 33+1 g-->a mutation, previously described in a Dutch family, was found in two patients; two novel mutations (33+2 t-->a, 33+5 c- >g) affected the two remaining patients. All the mutations resulted in the activation of a cryptic splice site 67 bp downstream in intron 1, leading to a frameshift and a premature stop codon in exon 4. Mutations in the exon 1 donor splice site are involved in eight of the nine non-erythroid variant AIP families reported in the literature. These data show that most mutations causing the non erythroid variant AIP are exon 1 splice defects, in contrast with classical AIP, where missense mutations are chiefly involved. Moreover, the allelic heterogeneity of PBGD gene defects causing the non-erythroid variant AIP is demonstrated, with five different mutations identified. These mutations could be easily detected by a single denaturing gradient gel electrophoresis which also allows the presymptomatic detection of gene carriers in the affected families. PMID- 9860300 TI - Change of bone mass in postmenopausal Caucasian women with and without hormone replacement therapy is associated with vitamin D receptor and estrogen receptor genotypes. AB - Our purpose is to assess whether genotypes of the vitamin D receptor (VDR) and estrogen receptor (ER) and their interaction influence changes in bone mass in postmenopausal Caucasian women with and without hormone replacement therapy (HRT). A population of 108 US Mid-West women who participated in a study of low dose continuous estrogen/progestin was genotyped at the VDR BsmI site and the ER XbaI and PvuII sites. Adequate vitamin D and calcium nutritional intakes were assured in all the study subjects. For the 3.5-year duration of the study, we analyzed changes in bone mineral density (BMD) at the spine, femoral neck, distal radius, and the total body (total body bone mineral content, tbBMC). We adjusted for confounding factors, such as age and weight, in the analysis. We found that VDR and/or ER genotypes and/or their interaction generally had significant effects on the changes in the bone mass measurements in both the placebo and HRT groups. When a significant gene-by-gene interaction exists between VDR and ER genotypes, failure to account for them in analyses may yield nonsignificant results, even if significant genotypic effects exist. The amount of variation in changes in bone mass measurements explained by the total genotypic effects of the VDR and ER loci varies from approximately 1.0% (for the tbBMC changes in combined placebo and HRT groups) to approximately 18.7% (for the spine BMD changes in the HRT group). These results suggest that individual genotypes are important factors in determining changes in bone mass in the elderly with and without HRT and thus may need to be considered with respect to the treatment to preserve bone mass in elderly Caucasian women. PMID- 9860302 TI - Complete physical map of the common deletion region in Williams syndrome and identification and characterization of three novel genes. AB - Williams syndrome (WS) is a contiguous gene deletion disorder caused by haploinsufficiency of genes at 7q11.23. We have shown that hemizygosity of elastin is responsible for one feature of WS, supravalvular aortic stenosis (SVAS). We have also implicated LIM-kinase 1 hemizygosity as a contributing factor to impaired visual-spatial constructive cognition in WS. However, the common WS deletion region has not been completely characterized, and genes for additional features of WS, including mental retardation, infantile hypercalcemia, and unique personality profile, are yet to be discovered. Here, we present a physical map encompassing 1.5 Mb DNA that is commonly deleted in individuals with WS. Fluorescence in situ hybridization analysis of 200 WS individuals shows that WS individuals have the consistent deletion interval. In addition, we identify three novel genes from the common deletion region: WS-betaTRP, WS-bHLH, and BCL7B. WS-betaTRP has four putative beta-transducin (WD40) repeats, and WS-bHLH is a novel basic helix-loop-helix leucine zipper (bHLHZip) gene. BCL7B belongs to a novel family of highly conserved genes. We describe the expression profile and genomic structure for each of these genes. Hemizygous deletion of one or more of these genes may contribute to developmental defects in WS. PMID- 9860301 TI - Alteration of the LIS1 gene in Japanese patients with isolated lissencephaly sequence or Miller-Dieker syndrome. AB - LIS1 is a genetic entity that is responsible for lissencephaly. Previously we have reported isolated lissencephaly sequence (ILS) in a Japanese patient carrying a balanced chromosomal translocation that disrupted the LIS1 gene. We examined mutations of LIS1 in 12 additional Japanese patients, 8 of them with ILS and 4 with Miller-Dieker syndrome (MDS). Fluorescence in situ hybridization (FISH) analysis disclosed deletions of part of the LIS1 gene or of the chromosomal region surrounding it in three of the ILS cases and in three of the MDS cases. In one of the remaining five ILS cases, SSCP analysis and subsequent sequence analysis identified a 1-bp deletion in exon IV, which can be expected to result in premature termination of the gene product. Our results indicate that in Japan, as elsewhere, abnormality of the LIS1 gene is a common cause of MDS/ILS. PMID- 9860303 TI - Neurofibromatosis pseudogene amplification underlies euchromatic cytogenetic duplications and triplications of proximal 15q. AB - Cytogenetically visible interstitial duplications of proximal 15q, which lack the Prader-Willi Angelman critical region (PWACR) frequently segregate in families without phenotypic effect, but the nature of the extra euchromatin has remained unclear. We used comparative genome hybridisation to confirm that the extra material in a cytogenetic triplication originated from proximal 15q. A PAC clone containing sequences specific for the type-1 neurofibromatosis (NF-1) pseudogenes, which map to 15q11.2, hybridised along the length of the enlarged region between the PWACR and the centromere. Computerised measurement of the fluorescent signal from the enlarged and normal chromosomes gave an average ratio of 9.85:1, consistent with amplification. In a second family, an amplified P1-4 signal co-segregated with a cytogenetic duplication and the average ratio between amplified and normal signals in the proband was 8.22:1. Ratios in noncarrier family members and control individuals were close to unity in most cases, but significantly greater than one in at least one instance. Our results provide a novel explanation for cytogenetic variation in 15q11.2. They also suggest that NF 1 pseudogene copy number may be polymorphic in the normal population, and that high copy numbers can produce G bands which do not reflect those of the normal constitutional karyotype. PMID- 9860304 TI - Association of infantile convulsions with paroxysmal dyskinesias (ICCA syndrome): confirmation of linkage to human chromosome 16p12-q12 in a Chinese family. AB - We have studied one family of Chinese origin, in which benign infantile convulsions and paroxysmal choreoathetosis (of the dystonic form) were co inherited as a single autosomal dominant trait. This association is specific to ICCA syndrome, which we have recently described in four French families. Some patients in the new family also exhibit recurrence of epileptic seizures at a much later age, making the ICCA syndrome in this family atypical. DNA samples isolated from this family of 22 members (9 affected) have been tested with genetic markers at chromosome 16p12-q12, in which region the ICCA syndrome has previously been linked. Confirmation of linkage to this pericentromeric region of human chromosome 16 has been obtained and no critical meiotic recombination event has been detected in the ICCA region. This result suggests that, in contrast to marked clinical heterogeneity, the association of infantile convulsions with paroxysmal dyskinetic movements could be genetically homogeneous. PMID- 9860305 TI - Molecular characterization of phenylketonuria in Japanese patients. AB - We characterized phenylalanine hydroxylase (PAH) genotypes of Japanese patients with phenylketonuria (PKU) and hyperphenylalaninemia (HPA). PKU and HPA mutations in 41 Japanese patients were identified by denaturing gradient gel electrophoresis and direct sequencing, followed by restriction fragment length polymorphism analysis to find a large deletion involving exons 5 and 6. Of 82 mutant alleles, 76 (92%) were genotyped showing 21 mutations. The major mutations were R413P (30.5%), R243Q (7.3%), R241 C (7.3%), IVS4nt-1 (7.3%), T2781 (7.3%), E6nt-96A-->g (6.1%), Y356X (4.9%), R111X (3.7%), and 442-706delE5/6 (2.4%). Eight new mutations (L52 S, delS70, S70P, Y77X, IVS3nt-1, A132 V, W187 C, and C265Y) and a polymorphism of IVS10nt-14 were detected. In vitro PAH activities of mutant PAH cDNA constructs were determined by a COS cell expression system. Six mutations, viz., R408Q, L52 S, R241 C, S70P, V388 M, and R243Q, had 55%, 27%, 25%, 20%, 16% and 10% of the in vitro PAH activity of normal constructs, respectively. The mean pretreatment phenylalanine concentration (0.83+/-0.21 mmol/l) of patients carrying the R408Q, R241 C, or L52 S mutation and a null mutation was significantly lower (P<0.0005) than that (1.99+/-0.65 mmol/l) of patients with both alleles carrying mutations associated with a severe genotype. Simple linear regression analysis showed a correlation between pretreatment phenylalanine concentrations and predicted PAH activity in 29 Japanese PKU patients (y=31.9-1.03x, r=0.59, P<0.0001). Genotype determination is useful in the prediction of biochemical and clinical phenotypes in PKU and can be of particular help in managing patients with this disorder. PMID- 9860306 TI - Identification of de novo chromosomal markers and derivatives by spectral karyotyping. AB - Despite major advances in molecular cytogenetics during the past decade and the important diagnostic role that fluorescence in situ hybridization (FISH) plays in the characterization of chromosomal abnormalities, the usefulness of this technique remains limited by the number of spectrally distinguishable fluorochromes or fluorochrome combinations. Overcoming this major limitation would allow one to use FISH to screen the whole human genome for chromosomal aberrations which, until recently, was possible only through conventional karyotyping. A recently described molecular cytogenetics technology, 24-color FISH using spectral karyotyping (SKY), permits the simultaneous visualization of all human chromosomes in 24 different colors. Most chromosomal aberrations detected during cytogenetic evaluation can be resolved using routine cytogenetic techniques alone or in combination with single- or dual-color FISH. However, some cases remain unresolved, in particular de novo supernumerary marker chromosomes and de novo unbalanced structural rearrangements. These findings cause particular diagnostic and counseling concerns when detected during prenatal diagnosis. The purpose of this report is to demonstrate the application of SKY in the characterization of these de novo structural chromosomal abnormalities. Eight cases are described in this report. SKY has considerable diagnostic applications in prenatal diagnosis because of its reliability and speed. The identification of the chromosomal origin of markers and unbalanced translocations provides the patient, physician, and genetic counselor with better predictive information on the phenotype of the carrier. PMID- 9860307 TI - FRAXE: the HindIII/OXE20 restriction polymorphism is not a rare variant. PMID- 9860308 TI - Mass screening for prostate cancer. PMID- 9860309 TI - Screening for prostate cancer--more questions than answers. PMID- 9860310 TI - A guinea pig's view on prostate cancer screening trials. PMID- 9860311 TI - Accelerated hyperfractionated radiotherapy combined with induction and concomitant chemotherapy for inoperable non-small-cell lung cancer--impact of total treatment time. AB - Tumour cell proliferation during conventionally fractionated radiotherapy (RT) can negatively influence the treatment outcome in patients with unresectable non small-cell lung cancer (NSCLC). Accelerated and hyperfractionated RT may therefore have an advantage over conventional RT. Moreover, earlier studies have suggested improved survival with addition of cisplatin-based chemotherapy (CT). We present here the results of combined treatment with induction and concomitant CT and accelerated hyperfractionated RT in a retrospective series of patients with advanced NSCLC. Between August 1990 and August 1995, 90 consecutive patients, aged 42-77 years (median 63 years), with locally advanced unresectable or medically inoperable NSCLC and good performance status were referred for treatment: stage: I 23%, IIIa 37%, IIIb 40%. Patient histologies included: squamous cell carcinoma 52%, adenocarcinoma 34% and large cell carcinoma 13%. The treatment consisted of two courses of CT (cisplatin 100 mg/m2 day 1 and etoposide 100 mg/m2 day 1-3 i.v.), the second course given concomitantly with RT. The total RT dose was 61.2-64.6 Gy, with two daily fractions of 1.7 Gy. A one-week interval was introduced after 40.8 Gy to reduce acute toxicity, making the total treatment time 4.5 weeks. Concerning toxicity, 33 patients had febrile neutropenia, 10 patients suffered from grade III oesophagitis and 7 patients had grade III pneumonitis. There were two possible treatment-related deaths, one due to myocardial infarction and the other due to a pneumocystis carinii infection. The 1-, 2- and 3-year overall survival rates were 72%, 46% and 34%, respectively; median survival was 21.3 months. Fifty-nine patients had progressive disease: 21 failed locoregionally, 29 had distant metastases and 9 patients had a combination of these. Pretreatment weight loss was the only prognostic factor found, except for stage. However, the results for stage IIIb were no different from those for stage IIIa. We conclude that the survival results compare favourably with those of most other studies with a manageable toxicity. PMID- 9860312 TI - Risk of breast cancer and changes in mammographic parenchymal patterns over time. AB - The relationship between sequential mammographic parenchymal patterns and breast cancer was estimated and the results were applied to selective screening. In a pilot screening program 4163 Finnish women aged 40-47 years at entry were invited to be screened every second year from 1982 to 1990. Mammographic parenchymal patterns (Wolfe's classification) were recorded at each screening round. The follow-up ended in 1993 and up until that time 68 new breast cancers were diagnosed. The age-adjusted relative risk of breast cancer was 2.5 (95% CI 1.5 4.0) among women with high-risk mammographic parenchymal patterns (P2,DY) at the screenings preceding cancer diagnosis compared with those with low-risk patterns (N1,P1). After further adjustment for body mass index, number of pregnancies and size of the breast, the relative risk increased to 2.8 (95% CI 1.7-4.9). The mammographic parenchymal pattern is an independent risk factor of breast cancer but not strong enough to be used as a criterion for selective screening. PMID- 9860313 TI - Radiotherapy in Scandinavia. PMID- 9860314 TI - Verification of single beam treatment planning using a ferrous dosimeter gel and MRI (FeMRI). AB - A method for analysing and comparing treatment planning system (TPS) data and ferrous dosimeter gel measurements evaluated with MRI (FeMRI) was developed, including image processing to final absorbed dose images. Measurements were analysed according to this method and FeMRI data were thereby compared with the TPS-calculated dose distribution. For photons, differences between FeMRI- and TPS dose data were mainly within +/- 2%. Minor shortcomings found in both the FeMRI system and the TPS are explained and discussed. For electron beams, there was an overall good agreement. It was found that the TPS underestimates the lateral scattering dose outside the primary beam, but the reported dose difference corresponds to a small spatial deviation (less than 2 mm). It is important to consider this single beam data comparison when the method is extended to more complicated situations, for example when using several beams. PMID- 9860315 TI - Tumor hypoxia and gene expression--implications for malignant progression and therapy. AB - Most histopathological classifications of human cancers include significant numbers of hypoxic cells. There is increasing evidence that, at least in certain types of human solid tumors, there is a positive relationship between the presence of hypoxia and poor outcome after radiation therapy alone or radiation combined with other therapies. Hypoxia appears to be an independent prognostic factor. There is evidence for enhanced malignant progression associated with hypoxia, including locoregional invasion and distant metastases. The presence of hypoxia may negatively affect outcome by induction of radiation resistance by the classical oxygen effect and/or by effects on gene expression and malignant progression, causing more aggressive locoregional and distant disease. It is now clear that hypoxia has the potential to influence expression of genes and activities of associated proteins that regulate growth and tissue homeostasis, resulting in cellular phenotypic heterogeneity. The molecular pathways involved in signaling and regulating changes in gene activities in response to external stresses such as hypoxia are becoming known. Identification of patients with hypoxic tumors will lead to improved selective therapy. PMID- 9860316 TI - Radiation-induced apoptosis--the ceramide-SAPK signaling pathway and clinical aspects. AB - Apoptosis, or programmed cell death is an important regulatory mechanism that is involved in a variety of homeostatic processes. Decreased cellular sensitivity or inappropriate responses to apoptotic stimuli may be important factors in tumorigenesis and resistance to anticancer treatments. It is generally accepted that all mammalian cells constitutively express the biochemical machinery to execute apoptosis. It is, however, not clear which signal transduction pathways are involved, or to which extent various stimuli activate independent or partially overlapping pathways. In this paper we discuss the involvement of a ceramide-mediated stress-activated protein kinase (SAPK) signaling cascade in radiation-induced apoptosis. Furthermore, examples are presented of pharmacological intervention in specific signal transduction pathways that lead to modulation of the apoptotic response. Finally, data are presented to illustrate the potential clinical relevance of apoptosis. PMID- 9860318 TI - Aspects on the development of radiation therapy and radiation biology since the early work of Rolf Wideroe. PMID- 9860317 TI - How to optimize therapeutic ratio in brachytherapy of head and neck squamous cell carcinoma? AB - Considerable experience has been accumulated with low dose rate (LDR) brachytherapy in the treatment of squamous cell carcinoma of the oral cavity and oropharynx, 4 cm or less in diameter. Recent analysis of large clinical series provided data indicating that modalities of LDR brachytherapy should be optimized in treating these tumours for increasing therapeutic ratio. LDR brachytherapy is now challenged by high dose rate (HDR) brachytherapy and pulsed dose rate (PDR) brachytherapy. Preliminary results obtained with the last two modalities are discussed in comparison with those achieved with LDR brachytherapy. PMID- 9860319 TI - Rolf Wideroe--Why is the originator of the science of particle accelerators so neglected, particularly in his home country? PMID- 9860320 TI - Rolf Wideroe and the development of particle accelerators. AB - The development of particle accelerators is traced from the simple but ingenious table-top devices conceived during the late 1920s to the present day's large, complex machines which extend over tens of kilometers. The emphasis is on Rolf Wideroe and his seminal contributions to the field. Not only did Wideroe construct the first accelerator which accelerated charged particles to an energy higher than the maximum voltage difference is the accelerator proper, he also invented particle colliders, today's work horse in experimental particle physics. PMID- 9860321 TI - Decreased CD11b expression, phagocytosis, and oxidative burst in urban particulate pollution-exposed human monocytes and alveolar macrophages. AB - Elevated levels of air pollution particulates < or = 10 microm in diameter (PM10) have been associated with an increase in mortality and morbidity due to pulmonary complications, including pneumonia. Impairment of inflammatory and host defense functions of the alveolar macrophage (AM) may be a precipitating factor. The present study was undertaken to determine whether human AM and blood derived monocytes (MO) modulate the expression of receptors important for phagocytosis of opsonized microbes (CD11b, CD11c), gram-negative bacteria (CD14), extracellular matrix interaction (CD29), and immune responses (CD11a, CD54, HLA-DR) when exposed to particulates obtained from urban air (UAP). Furthermore, phagocytosis of and oxidant generation by opsonized yeast were investigated in particle exposed cells. AM and MO exposed to UAP for 18 h expressed significantly lower levels of CD11b and CD29. CD14 expression was markedly decreased in MO but not in AM, and CD11c was reduced in AM but not in MO. CD11a, CD54, and HLA-DR were unaltered in both phagocyte populations. Decreased receptor expression was not dependent on particle load in the cells. Phagocytosis of Saccharomyces cerevisiae and the chemiluminescence response were also significantly inhibited by UAP. Time course studies revealed that decreased oxidant generation was evident already at 3 h postexposure, while significant effects on phagocytosis and CD11b expression were found at 18 h. These data indicate that exposure to particulate pollution is likely to impair host defense functions of AM and MO, which are important in elimination of a variety of pathogens in the lung. PMID- 9860322 TI - Effects of in vitro exposure of beluga whale leukocytes to selected organochlorines. AB - The effects of in vitro exposure to different organochlorines were evaluated on immune functions of beluga whale peripheral blood leukocytes and splenocytes. The effects of different concentrations of four different congeners of PCBs (138, 153, 180, and 169) as well as two DDT metabolites (p,p'-DDT and p,p'-DDE) were evaluated on phagocytosis and cell proliferation. The effects of dioxins and mixtures of organochlorines were also evaluated on cell proliferation. The different compounds tested had no marked effect on phagocytosis. PCB 138 and p,p' DDT, but not PCB 153, PCB 180, PCB 169, and p,p'-DDE, reduced significantly the proliferative response of beluga splenocytes cultured either with or without phytohemagglutinin A (PHA). Proliferation of beluga splenocytes was not markedly affected by exposure to 5 ppm of PCB 138, 153, 180, and 169 separately. Exposure to a mixture of congeners 138, 153, and 180 (5 ppm each) significantly reduced splenocytes proliferation, but not the mixture of congeners 138, 153, 180, and 169 (5 ppm each). TCDD did not affect cell proliferation in our study. The reduced proliferation of beluga cells exposed in vitro to mixtures of organochlorines at concentrations in the range of those observed in tissues of St. Lawrence belugas might provide a basis to support the hypothesis that contaminants induce immunosuppression in these animals. PMID- 9860323 TI - Effects of lead on behavior, growth, and survival of hatchling slider turtles. AB - In this study the effects of lead on behavioral development of hatchling slider turtles (Trachemys scripta) from the Savannah River Site, near Aiken, SC, were examined. It was of interest to determine whether dose or size affects survival, growth, or behavior. Hatchlings from 1995 showed no significant differences in growth, survival, or behavior between control and lead-injected animals at a dose of 0.05 and 0.1 mg/g (n = 10 per group). In 1996, 48 hatchlings were divided into four groups injected with 0 (control), 0.25, 1, or 2.5 mg/g lead. Few significant differences occurred in growth or size as a function of lead treatment at 4 mo of age, but survival declined markedly as a function of lead dose. Righting response was significantly impaired by lead; time to right was directly related to lead dose. Size also affected behavior; larger hatchlings turned over more quickly and reached cover sooner than did smaller hatchlings. PMID- 9860324 TI - Effect of polycyclic aromatic hydrocarbons on the elimination kinetics of pyrene and the urinary excretion profile of 1-hydroxypyrene in the rat. AB - Pyrene was chosen as a noncarcinogen model of polycyclic aromatic hydrocarbons (PAHs). Groups of male Wistar rats were dosed with pyrene and with mixture of pyrene and fluoranthene, pyrene and benz[a]anthracene, or pyrene, fluoranthene, and benz[a]anthracene at 20 mg/kg by intravenous or oral routes. Blood samples were taken at 0.25, 0.5, 1, 2, 3, 4, and 5 h after administration. The concentration of pyrene was determined by gas chromatography. The toxicokinetic parameters for pyrene were determined from the time course of blood concentration. A significant increase in the bioavailability of pyrene after treatment with other PAHs was observed. Urinary 1-hydroxypyrene excretion was analyzed after pretreatment with acenaphthene, naphthalene, chrysene, phenanthrene, benz[a]anthracene, and benzo[a]pyrene. The urine from rats was collected for 3 d and the concentration of 1-hydroxypyrene was determined using high-performance liquid chromatography (HPLC). Most compounds examined caused a decrease in the urinary excretion of the metabolite of pyrene. PMID- 9860325 TI - Aflatoxin B1-induced suppression of nitric oxide production in murine peritoneal macrophages. AB - Aflatoxin B1 (AFB1), a potent hepatocarcinogen, is known to impair specific and non-specific immune responses. AFB1 mainly decreases lymphocyte functions and may also affect macrophages assisting lymphocyte functions. Macrophages play an important role in a host defense against tumors and bacteria. Furthermore, some macrophage products, including nitric oxide (NO), may be involved in cytotoxicity. The effect of aflatoxin B1 (AFB1) was investigated on NO production from murine peritoneal macrophages. Macrophages were pretreated with AFB1 for 24 h and then stimulated with lipopolysaccharide (LPS) for 24 h. AFB1 at 10 or 50 microM reduced the production of NO. Compared to vehicle control, there was a greater reduction of NO production with increased AFB1 pretreatment and LPS stimulation. AFB1 at 10 or 50 microM decreased inducible nitric oxide synthase (iNOS) activity about 24% and 28%, respectively, after stimulation with 1 microg/ml LPS and about 12% and 24%, respectively, after stimulation with 10 microg/ml LPS. AFB1 pretreatment also decreased the synthesis of iNOS protein and the mRNA of macrophages. Taken together, these results suggest that AFB1 pretreatment reduces NO production from murine peritoneal macrophages stimulated by LPS, which is mediated by the reduction of iNOS activity, mRNA, and protein. PMID- 9860326 TI - Staging of colorectal cancer: biology vs. morphology. AB - PURPOSE: An accurate determination of the extent or staging of a disease is critical, because it provides the basis for making therapeutic decisions. Staging is a collaborative effort by the surgeon and the pathologist. Radioimmunoguided surgery has been evaluated for its ability to help surgeons determine the extent of disease during surgery, when management decisions have the most impact on patient care. This study was done to compare radioimmunoguided surgery "biostaging" with traditional pathologic staging (TNM) as predictors of survival in patients undergoing curative resections for colorectal cancer. METHODS: Ninety seven patients with colorectal cancer were prospectively enrolled in radioimmunoguided surgery protocols. Evaluation of follow-up survival data was performed. All patients underwent exploratory laparotomy and radioimmunoguided surgery with resection of their primary colorectal tumor. Survival data were analyzed with the Kaplan-Meier method with log-rank comparisons. RESULTS: Of 97 patients enrolled in the study, 59 were evaluable and completely resectable by radioimmunoguided surgery. Mean follow-up was 62 months, with a range of 34 to 89 months. By traditional staging 13 patients were pStage I, 18 patients were pStage II, and 28 patients were pStage III. By radioimmunoguided surgery biostaging, 24 patients were radioimmunoguided surgery-negative whereas 35 patients were radioimmunoguided surgery-positive. Survival rates by pathologic stage approached a significant difference, but did not, as of the conclusion of the study period, reach it (P = 0.12). Survival rates based on radioimmunoguided surgery status demonstrated a highly significant difference (P = 0.0002). CONCLUSIONS: Radioimmunoguided surgery biostaging provides new information intraoperatively on cancer staging that has not been available before. This may lead to new strategies for therapy that can be individualized and optimized for each patient with cancer. PMID- 9860327 TI - Results of salvage abdominoperineal resection for anal cancer after radiotherapy. AB - PURPOSE: Nonsurgical treatment of anal cancer by radiotherapy alone or combined with chemotherapy is the standard therapy for epidermoid carcinoma of the anal canal. Surgery is only recommended for treatment failures. Very few studies have been devoted to the outcome of this salvage surgery. The aim of this study is to evaluate these results. METHODS: A retrospective review from 1986 to 1995 revealed 21 patients with residual or recurrent anal canal carcinoma after initial radiotherapy, operated on by abdominoperineal resection. Patients were reviewed as to age, gender, initial treatment, any symptoms of recurrence, duration until recurrence, any diagnosis imaging, treatment, and outcome. RESULTS: None of these 21 patients had known lymph node involvement or metastases at radiotherapy or at salvage abdominoperineal resection. Eleven patients had residual disease (positive biopsy less than 6 months after the end of radiotherapy) and 10 had tumor recurrence (more than 6 months after cessation of treatment). Recurrence occurred at a mean of 15 (range, 9-41) months after radiotherapy. All 21 patients underwent an abdominoperineal resection. Pathologic examination of the 21 specimens showed complete excision in all cases except one and lymph node metastases in two cases. There was no perioperative mortality. The mean follow-up after surgery was 40 months; no patients were lost to follow-up. Of the 21 patients, 10 died and 11 lived, of whom 9 are disease free. The overall survival rate at three years after salvage abdominoperineal resection was 58 percent. The overall survival rate for patients with residual disease (vs. recurrence) at three years was 72 percent (vs. 29 percent) and at five years was 60 percent (vs. 0 percent; P = 0.06). CONCLUSIONS: Salvage abdominoperineal resection for anal cancer can be expected to yield a number of survivors from residual disease, but the low rate of survival after abdominoperineal resection for recurrent disease suggests the need for additional postoperative treatment if salvage abdominoperineal resection is performed. PMID- 9860328 TI - Different prognostic effect of postoperative chemoradiation therapy on diploid and nondiploid high-risk rectal cancers. AB - PURPOSE: DNA ploidy has been shown to play a role in the response to cytotoxic therapy in a variety of malignancies, including breast cancer and melanoma. However, the importance of DNA ploidy in rectal cancer is unknown. The aim of the present study was to determine whether ploidy status might be associated with response to postoperative chemoradiation in TNM Stages II to III rectal cancer. METHODS: This retrospective study analyzed data from 229 patients with TNM Stages II to III rectal cancer who underwent resection between 1979 and 1984. The ploidy status and treatment modalities in relation to outcome were assessed. RESULTS: The recurrence-free ten-year survival rate was 52.2 percent for patients with diploidy and 50.5 percent for patients with nondiploidy (P = 0.99). The ten-year survival rates for patients with diploidy and patients with nondiploidy were 55 and 19 percent (P = 0.016) in the chemoradiation group, and 51 and 60 percent (P = 0.15) in the nonchemoradiation group, respectively. In the chemoradiation group, DNA nondiploidy was associated with an increased recurrence rate (83.3 vs. 50.0 percent; P = 0.001). The interaction between DNA nondiploidy and chemoradiation remained important in predicting outcome in the Cox regression model. Factors independently correlated with a worse outcome included Stage IIIb (relative risk, 2.9; 95 percent confidence interval, 1.7-5; P = 0.0001), perineural invasion (relative risk, 2.5; 95 percent confidence interval, 1.6-4, P = 0.0001), distal tumor (relative risk, 1.7; 95 percent confidence interval, 1.1 2.7, P = 0.014), and nondiploidy with chemoradiation (relative risk, 2.9; 95 percent confidence interval, 1.2-7.2, P = 0.0213). CONCLUSIONS: These findings suggest that DNA nondiploidy is inversely correlated with long-term outcome among patients with high-risk rectal cancer receiving chemoradiation. PMID- 9860329 TI - Cecal intubation model in the rat that facilitates selective in vivo study of colonic epithelial biology: preliminary report. AB - PURPOSE: A free-living animal model with ready and repetitive access to selected regions of the large bowel and with minimally altered bowel anatomy and physiology would facilitate the in vivo study of luminal factors on the colonic mucosa in a steady-state environment. This study describes a novel model of large bowel intubation in the rat. METHOD: Four Sprague-Dawley rats (240-260 g) had laparotomy and intubation of the distal colon and the cecum via a cecotomy with the use of two small tubes with restraints and transmural anchors. The tubes were tunneled and anchored to the back for infusion of fluid directly into the colon. Tube positions were studied when the animals were killed. Animals were fed on either a 10 percent fiber diet or a fiber-free diet. Stathmokinetic assessment of the distal colon was performed after one week of infusion with phosphate-buffered saline and sodium n-butyrate. RESULTS: The technique produced an easy access without affecting the weight gain of the animals after recovery. Tube positions were accurate after three weeks at the time the animals were killed. Infusions of phosphate-buffered saline and n-butyrate were well tolerated. n-Butyrate infusions twice daily for a week reversed the atrophy in the colonic mucosa induced by dietary fiber deprivation. CONCLUSION: An in vivo large-bowel intubation model permitting selective delivery of luminal factors provides an effective option for the study of colonic mucosal biology. PMID- 9860330 TI - Model of implantation of tumor cells simulating recurrence in colonic anastomosis in mice. AB - PURPOSE: Local recurrence after colorectal cancer surgery is usually perianastomotic. An experiment was designed to investigate whether free intraluminal cells can penetrate through a colonic anastomosis and thereby cause local recurrence. METHODS: BALB/c and C57/BL mice underwent ascending colotomy followed by watertight anastomosis. Thereafter, CT-26 murine colon carcinoma cells were injected into the cecal lumen 2 cm proximal to the anastomosis of syngeneic BALB/c mice, whereas B-16 murine melanoma cells were injected in the same fashion into C57/BL mice. Control animals without anastomosis received similar injections. Animals were killed 24 hours, 72 hours, and 30 days after surgery and were checked for tumorigenesis. RESULTS: Results of peritoneal fluid cytology were negative after 24 hours, whereas after 72 hours cancer cells were identified in the peritoneal fluid of 80 percent of mice with colotomy and anastomosis compared with 20 percent of control mice. Thirty days after surgery, 11.1 percent of the control BALB/c mice developed pericecal tumor growth, similar to the overall rate of murine melanoma in C57/BL. In mice with anastomoses, perianastomotic tumor growth was observed in 47.5 percent of BALB/c mice (P < 0.001) and was correlated with the number of injected cells. Tumor growth reached approximately 75 percent tumor take with high cell densities, whereas in C57/BL mice no difference was found between the experimental and control groups. CONCLUSIONS: The findings suggest that free intraluminal cancer cells of colonic origin may penetrate through watertight anastomoses and implant on the anastomotic or peritoneal surface and initiate tumor growth. This anastomotic penetration is cell-mass dependent. The reported experimental model is simple, reproducible, and advantageous for studies of colonic anastomosis. PMID- 9860331 TI - Fulminant colitis in inflammatory bowel disease: detailed pathologic and clinical analysis. AB - PURPOSE: The morphologic features of fulminant colitis may be nonspecific, making differentiation between ulcerative colitis and Crohn's disease difficult, even after colectomy. The aims of this study were 1) to identify histologic features that accurately differentiated ulcerative colitis, Crohn's disease, and indeterminate colitis in fulminant colectomy specimens; 2) to determine how frequently subsequent clinical course altered the pathologic diagnosis; and 3) to evaluate the natural history of histologically diagnosed indeterminate colitis. METHODS: Ninety-five fulminant colectomy specimens were evaluated, of which 85 had an original diagnosis of fulminant inflammatory bowel disease. Complete pathologic material and comprehensive clinical follow-up information was available on 67 cases of inflammatory bowel disease. These were re-evaluated in a blinded fashion, and histopathologic features were compared with the original diagnosis and reviewed in the light of subsequent clinical behavior to reach a final diagnosis. RESULTS: Evaluation of macroscopic features was not helpful in differentiating ulcerative colitis from Crohn's disease. Microscopic examination correctly diagnosed ulcerative colitis or Crohn's disease in only 58 of 67 (87 percent) cases. A further three cases (4 percent) were definitively classified after correlation with clinical data, leaving a residual six cases that were diagnosed as indeterminate colitis. Granulomas and lymphoid aggregates were the two most specific indicators of Crohn's disease. CONCLUSIONS: Histopathologic evaluation alone has limitations in the accurate classification of fulminant inflammatory bowel disease. Histologically diagnosed indeterminate colitis is a heterogeneous group that may include some patients who subsequently prove to have ulcerative colitis or Crohn's disease. PMID- 9860332 TI - Pudendal neuropathy is predictive of failure following anterior overlapping sphincteroplasty. AB - PURPOSE: This study assessed the efficacy of anterior overlapping sphincteroplasty and parameters predictive of a successful outcome. METHODS: Clinical findings and physiologic investigations of female patients who underwent anterior overlapping sphincteroplasty for fecal incontinence between 1988 and 1996 were reviewed. The extent of sphincter damage was assessed at needle electromyography as the number of quadrants exhibiting decreased motor unit potentials. Prolonged pudendal nerve terminal motor latencies were those of greater than 2.2 ms. The size of the endoanal ultrasound defect was assessed as degrees circumference of the external sphincter in which viable muscle was absent. Patients were reviewed by telephone questionnaire and were asked to grade the outcome of their surgery as excellent or good (success) or fair or poor (failure). Incontinence was graded using a scoring system of 0 (perfect continence) to 20 (complete incontinence). RESULTS: There were 100 patients who had an overlapping sphincteroplasty; complete follow-tip information was obtained for 77 patients at a median of 24 (range, 2-96) months. The median age was 47 (range, 25-80) years and they had a median duration of incontinence of four (range, 0.1-39) years. Prior sphincteroplasty had been performed in 30 patients with a median of one (range, 1-7) operations. Investigations performed included electromyography (n = 49), pudendal nerve terminal motor latency (n = 71), endoanal ultrasound (n = 49), and manometry (n = 67). Sixty percent of patients had improved continence and 42 (55 percent) considered their surgery to have been successful as attested to by a significant decrease in their incontinence score (from 15.1 +/- 4.5 to 4.3 +/- 4.2; P < 0.0001). Neither patient age, parity, prior sphincteroplasty, cause or duration of incontinence, extent of electromyography damage, size of the endoanal ultrasound defect, nor any manometric parameter correlated with outcome. However, 62 percent of 59 patients with bilaterally normal pudendal nerve terminal motor latencies had a successful outcome compared with only 16.7 percent of 12 patients with unilateral or bilateral prolonged pudendal nerve terminal motor latencies (P < 0.01). CONCLUSION: Bilateral normal pudendal nerve terminal motor latencies are the only factors predictive of long-term success after overlapping sphincteroplasty. PMID- 9860333 TI - Natural history of diverticular disease: when to operate? AB - PURPOSE: The natural history of patients admitted because of acute diverticulitis is largely unknown, and the selection of patients for surgical treatment varies notably. This study presents our experience concerning the outcome for 366 patients admitted during a 10-year period. METHODS: Three hundred sixty-six patients admitted to our hospital with acute diverticulitis from 1981 to 1990 were identified from a computer database, and their clinical data up to the end of 1996 were reviewed from the database and patient records. RESULTS: There were significantly more males than females in the age group less than 50 years old, and young males underwent surgical treatment during the first treatment period more frequently than the others. Young patients were operated on without mortality, and all their temporary colostomies were closed. Older patients died more often of diseases unrelated to the diverticular disease during the years after the first episode of acute diverticulitis. Recurrences of diverticular disease developed in 22 percent of patients, and they were significantly more common in patients less than 50 years old than in the older age groups. Males less than 50 years old more often developed complications of diverticular disease after two hospital admissions. CONCLUSIONS: Males first admitted when less than 50 years of age undergo more primary operations and develop more recurrences of diverticular disease than do older people. Based on our data, however, we recommend surgery for all patients after two episodes of acute diverticulitis that resolves after conservative treatment with antibiotics. PMID- 9860334 TI - Does HIV status influence the anatomy of anal fistulas? AB - PURPOSE: Although anorectal disease is common in human immunodeficiency virus positive patients, little is known about the type and anatomic distribution of anal fistulas in this patient group. The aim of this study was to compare anatomic characteristics of anal fistulas in human immunodeficiency virus positive patients with those in human immunodeficiency virus-negative patients by use of a retrospective chart review. METHODS: The charts of 146 male patients younger than 50 years with an anal fistula were reviewed. Incomplete fistulas referred to those tracts arising from an internal opening into either a blind sinus or an undrained abscess cavity. RESULTS: There were 60 human immunodeficiency virus-positive patients and 86 human immunodeficiency virus negative patients. Mean age of the human immunodeficiency virus-positive patient group was 37 years vs. 40 years for the human immunodeficiency virus-negative patient group. Thirty-one human immunodeficiency virus-positive patients (52 percent) were classified as having AIDS, and the remaining 29 patients (48 percent) were asymptomatic. Mean T helper cell count in the human immunodeficiency virus-positive patient group was 277 cells per microliter. Fistulous tracts were intersphincteric (n = 56), transsphincteric (n = 41), suprasphincteric (n = 2), and incomplete (n = 47). Incomplete fistulas were identified in 33 (55 percent) human immunodeficiency virus-positive patients vs. 14 (16 percent) human immunodeficiency virus-negative patients (P < 0.001). Of the 47 incomplete fistulas, 37 (79 percent) were found in association with an abscess cavity. All ten patients with an incomplete fistula into a blind sinus were human immunodeficiency virus-positive. The incidence of an incomplete fistula without an abscess was significantly higher in the human immunodeficiency virus-positive patient group (17 percent) compared with the human immunodeficiency virus-negative patient group (0 percent; P < 0.001). CONCLUSIONS: Anal fistulas in HIV-positive patients arise from the dentate line in similar locations to human immunodeficiency virus negative patients. However, human immunodeficiency virus-positive patients were more likely to have incomplete anal fistulas than human immunodeficiency virus-negative patients. Furthermore, human immunodeficiency virus-positive patients are predisposed to incomplete fistulas leading into a blind sinus. PMID- 9860335 TI - Associations between hemorrhoids and other diagnoses. AB - PURPOSE: The risk factors and mechanisms that contribute to the occurrence of hemorrhoids are not well understood. The study of the comorbid occurrences of hemorrhoids with other diagnoses in identical patients may point to a common underlying pathophysiology. The present study was undertaken to determine which diagnoses are associated with the occurrence of hemorrhoids. METHODS: A case control study compared the occurrence of comorbid diseases in case subjects with hemorrhoids with that of control subjects without hemorrhoids. The case population comprised all patients with hemorrhoids (International Classification of Diseases codes 455.0-455.9), who were discharged from hospitals of the U.S. Department of Veterans Affairs between 1986 and 1996. In a multiple logistic regression analysis, the occurrence of hemorrhoids served as outcome variable, and age, gender, ethnicity, and the comorbid occurrence of other diagnoses served as predictor variables. RESULTS: A total of 96,314 individual patients with hemorrhoids and the same number of control subjects were identified. In a chart review of a random sample of 100 cases, the diagnosis of hemorrhoids could be confirmed in 97 percent of all instances checked. The variety of diagnoses associated with hemorrhoids could be broken down into five large categories: 1) diseases associated with diarrhea (odds ratio, 1.30; 95 percent confidence interval, 1.27-1.33); 2) spinal cord injuries (odds ratio, 1.17; 95 percent confidence interval, 1.09-1.26); 3) constipation and related diseases (odds ratio, 1.48; 95 percent confidence interval, 1.43-1.54); 4) various types of anorectal diseases (odds ratio, 4.71; 95 percent confidence interval, 4.44-5.0); and 5) conditions that could be considered manifestations or sequelae of the hemorrhoidal disease itself (odds ratio, 3.41; 95 percent confidence interval, 3.30-3.51). CONCLUSIONS: The types and spectrum of comorbid diagnoses associated with hemorrhoids suggest that an increased tone of the anal sphincter constitutes a common pathophysiologic mechanism for the development of hemorrhoids. PMID- 9860336 TI - Cytokine production from colonic T cells in patients with ulcerative colitis with and without primary sclerosing cholangitis. AB - PURPOSE: Only five percent of all patients with ulcerative colitis develop primary sclerosing cholangitis. T cells accumulate at the sites of the colonic and bile duct inflammation in both ulcerative colitis and primary sclerosing cholangitis. T helper cell populations comprise functionally distinct subsets characterized by the cytokines they produce. Several alterations in cytokine production have been described in patients with ulcerative colitis. The aim of this study was to investigate possible differences in T helper subsets and cytokine production in peripheral blood and colonic mucosa among ulcerative colitis patients with and without primary sclerosing cholangitis. METHODS: Eleven patients with primary sclerosing cholangitis and extensive ulcerative colitis, 11 patients with extensive ulcerative colitis and no liver disease, and 5 patients without any history of liver disease who underwent routine colonoscopy because of previous polypectomy were included in the study. Colonoscopy with multiple biopsies was performed on all patients. Lamina propria mononuclear cells and peripheral blood mononuclear cells were isolated. A modified version of solid phase enzyme-linked immunospot assay was used for the separate counting of cells producing interferon-gamma, interleukin-2 (T helper 1), and interleukin-4 (T helper 2). RESULTS: No differences in spontaneous production of cytokines from peripheral blood mononuclear cells was found among the three groups. Patients with primary sclerosing cholangitis compared with patients with ulcerative colitis without liver disease showed a significant increase in the number of cells secreting interferon-gamma after purified protein derivative stimulation (P < 0.02). More cells secreting interferon-gamma were found in the two ulcerative colitis groups than in the cell populations from healthy controls (P < 0.03). The number of cells secreting interferon-gamma in the primary sclerosing cholangitis group was significantly lower than in the ulcerative colitis group without liver disease (P < 0.04). The number of cells secreting interleukin-4 was lower in the primary sclerosing cholangitis group than among the patients with ulcerative colitis only (P = 0.05). CONCLUSION: Isolated lymphocytes from colonic mucosa differ in cytokine production in patients with ulcerative colitis with and without primary sclerosing cholangitis. PMID- 9860337 TI - Intact colonic motor response to sudden awakening from sleep in patients with chronic idiopathic (slow-transit) constipation. AB - PURPOSE: There are few data about the relationships between colonic motor behavior and higher brain functions, such as sleep. Previous studies were done in healthy subjects, and it is unknown whether patients with functional motor disorders of the colon behave differently. This study was designed to characterize colonic motor activity in patients with constipation, both during sleep and after sudden awakening, and to compare it with that of healthy subjects. Our working hypothesis was that patients with constipation would have an impaired response to sudden awakening. PATIENTS AND METHODS: Twelve chronically constipated women, 22 to 49 years old, were recruited for the study, and their data were compared with those obtained from 12 healthy female volunteers, 21 to 38 years old. Manometric studies were performed in the descending and sigmoid colon for 30 minutes during sleep (immediately before awakening) and 30 minutes after being awakened suddenly. A motility index was calculated before and after the stimulus. RESULTS: In both groups motility in the descending and the sigmoid colon was almost absent during sleep and significantly increased after sudden awakening. No difference in postawakening values was found between patients with constipation and controls. CONCLUSIONS: In patients with chronic constipation, the brain-gut control of some fundamental mechanisms governing colonic motility is preserved. These data suggest that the alterations of colonic motility described in chronic constipation may be caused by an intrinsic dysfunction of the viscus. PMID- 9860338 TI - Long pedunculated colonic polyp composed of mucosa and submucosa: proposal of a new entity, colonic muco-submucosal elongated polyp. AB - We encountered 15 patients with colonic polyps showing histologic features that did not belong to any of the known categories. All polyps were elongated and drumstick-shaped, with lengths of 12 to 160 (mean, 29 mm) mm. Histologically, the polyps were covered with normal mucosa and consisted of edematous, loose, fibrous, connective tissues and dense, fibrous submucosal layers, often showing dilation of blood vessels and lymphatics. Although the mechanism of generation of such polyps remains unknown, their elongation may be caused by intestinal motion. Because this kind of polyp has not been described previously outside Japan, we here introduce a new type of polyp, which we have proposed calling the colonic muco-submucosal elongated polyp. PMID- 9860339 TI - Complications of abdominal stoma surgery. AB - PURPOSE: This study was undertaken to review and summarize the complications of ileostomy and colostomy creation and subsequent closure. METHODS: The English language medical literature for at least the past 15 years was reviewed comprehensively. RESULTS: Complications of surgery for the creation of end, loop, and "end loop" stomas are presented. Technical factors, which might influence complication rates, are discussed. Optimal management of ostomy complications is presented, especially for peristomal hernias. Similarly, techniques and complications for stoma closure are analyzed. CONCLUSIONS: Stoma creation is not a trivial undertaking; careful surgical technique minimizes complications (which are relatively frequent), and promotes good ostomy function. Peristomal hernias are difficult to cure permanently. The morbidity of ileostomy and colostomy closure is also appreciable. PMID- 9860340 TI - Surgery for intestinal graft-versus-host disease: report of two cases. AB - PURPOSE: With allogeneic bone marrow transplantation becoming increasingly common, intestinal graft-versus-host disease often is seen within specialized transplant units. Surgeons are involved infrequently in the management of such patients, and yet occasionally are called upon to operate on critically ill patients with refractory disease. This article reviews the salient features of this condition and discusses possible indications for surgery. METHOD: Two cases of intestinal graft-versus-host disease that required laparotomy are presented. RESULTS: A 24-year-old male with severe, unremitting, acute intestinal graft versus-host disease underwent small-bowel and large-bowel resections for massive bleeding. A 35-year-old male with recurrent symptoms of small-bowel obstruction after allogeneic bone marrow transplantation underwent small-bowel resection and multiple strictureplasties for chronic intestinal graft-versus-host disease. CONCLUSION: Surgery can be performed for complications of intestinal graft-versus host disease not responding to medical therapy, with successful outcome. PMID- 9860341 TI - Conservative surgical management of perforating Crohn's disease: side-to-side enteroenteric neoileocolic anastomosis: report of two cases. AB - Conservative surgical management of jejunoileal Crohn's disease is the first choice treatment in plurioperated patients with high risk of small-bowel syndrome. Treatment is more controversial in patients with a limited disease and no previous surgery, especially in those with terminal ileitis. Even in those cases we advocate conservative surgical management because we have demonstrated regression of both symptoms and morphologic lesions. Contraindications to nonresectional surgery include the presence of abscesses and fistulas. Impending short-bowel syndrome represents a partial exception to this approach. In this article two cases of conservative surgery performed in plurioperated patients with perianastomotic perforating recurrent disease are presented. We have named this procedure "side-to-side enteroenteric neoileocolic anastomosis," which is a combination of small resections and conservative procedure. This operative strategy leads us to believe that nonresectional surgery could be selectively performed even in patients with perforating Crohn's disease. PMID- 9860342 TI - Fungal sacral osteomyelitis as the initial presentation of Crohn's disease of the small bowel: report of a case. AB - We report a unique case of Candida albicans sacral osteomyelitis in a 48 year-old female with previously undiagnosed Crohn's disease. The patient was ill for one year with fatigue, weakness, and a 60-lb weight loss. At the time of presentation, she developed chills, fever, right lower quadrant abdominal pain, and right knee pain. Physical examination was significant for a palpable right lower quadrant abdominal mass. A computed tomographic scan of the abdomen and pelvis identified a large right-sided retroperitoneal mass, severe right hydronephrosis, and air within the right sacrum. Findings at laparotomy included small-bowel changes consistent with Crohn's disease, a multiloculated retroperitoneal abscess, and evidence of sacral osteomyelitis. A right hemicolectomy with sacral debridement and placement of presacral drains was performed. Bone cultures from the sacrum demonstrated a predominance of C. albicans, in addition to coliforms and enterococcus. The patient was placed on amphotericin B and intravenous antibiotics. Because serial computed tomographic scans of her pelvis demonstrated progression of her pelvic osteomyelitis to include the sacrum, right ilium, right acetabulum, and right femoral head, a repeat debridement with resection of the right femoral head was performed. After 12 months of follow-up, she was doing well without medications and had no constitutional symptoms or radiographic evidence of disease progression. This report illustrates a unique case of Crohn's disease presenting as sacral osteomyelitis secondary to small-bowel fistulization. Aggressive multidisciplinary surgical and medical management were the key to the successful management of this difficult case. PMID- 9860343 TI - Cecal infarct: report of a case. AB - PURPOSE: The purpose of this article is to present a case of an uncommon opportunistic fungal infection that appears in immunologically or metabolically compromised patients and is usually fatal. METHOD: A 54-year-old woman with an acute lymphoblastic leukemia had acute abdominal pain with peritoneal symptoms during her hospital stay. A laparotomy was performed and a cecal infarct firmly adherent to about 30 cm of infiltrated jejunal loop was discovered. RESULTS: Microscopic studies revealed an infiltration of the jejunal wall by abnormal lymphocytes. The cecal and ileal walls were infiltrated by leukocytes. Blood vessels were invaded by giant cells and large, unseptated, right-angle branching hyphea. CONCLUSIONS: These findings were considered typical of invasive mucormycosis of the cecum and the terminal ileum. PMID- 9860344 TI - Use of a jejunal pouch with ileal interposition in salvage surgery after restorative proctocolectomy. AB - PURPOSE: A novel technique is described for pouch reconstruction after failed restorative proctocolectomy and pouch excision. METHODS: Surgery was undertaken in two patients who had undergone restorative proctocolectomy with subsequent excision of the ileal J-pouch after necrosis. At revisional surgery it was technically impossible to form a pouch using the terminal ileum because of mesenteric shortening. A new 18-cm J-pouch was formed with a jejunal segment. After selective division of axial vessels, adequate length was obtained to allow formation of a jejunal-pouch-anal anastomosis. The small bowel distal to the pouch was interposed between the proximal jejunum and J-pouch and a defunctioning stoma was made. RESULTS: The postoperative course was uneventful in both cases. The functional results at 3 and 12 months after stoma closure were good, with five to seven bowel movements per day and complete continence. CONCLUSION: Shortening of the terminal ileal mesentery may preclude the formation of an ileal pouch in patients undergoing salvage surgery after failed restorative proctocolectomy. This novel technique of jejunal J-pouch formation and small bowel interposition has value as an alternative to definitive ileostomy or Kock's pouch in such patients. PMID- 9860345 TI - Factors involved in the maintenance of endothelial function. PMID- 9860346 TI - The role of tissue angiotensin-converting enzyme (ACE): studies of ACE mutant mice. PMID- 9860347 TI - Potential mechanisms for the effect of angiotensin-converting enzyme inhibitors on endothelial dysfunction: the role of nitric oxide. PMID- 9860348 TI - Bradykinin and the therapeutic actions of angiotensin-converting enzyme inhibitors. PMID- 9860349 TI - Impact of endothelin-1 on vascular structure and function. PMID- 9860350 TI - Effect of the angiotensin-(1-7) peptide on nitric oxide release. PMID- 9860351 TI - Hypertension, heart failure, and endothelial function. PMID- 9860352 TI - The effects of angiotensin-converting enzyme inhibition on endothelial dysfunction: potential role in myocardial ischemia. PMID- 9860353 TI - Primary and secondary prevention of coronary artery disease: trials of lipid lowering with statins. PMID- 9860354 TI - Effects of aspirin on endothelial dysfunction in atherosclerosis. PMID- 9860355 TI - Effects of antihypertensive agents on endothelial dysfunction: rationale for the brachial artery normalization of forearm function study. PMID- 9860356 TI - Cardioprotective effects of postmenopausal hormone treatment. PMID- 9860357 TI - Cardiovascular effects of soybean phytoestrogens. PMID- 9860358 TI - Nutriceuticals for cardiovascular health. PMID- 9860359 TI - Overview of angiotensin II-receptor antagonists. PMID- 9860360 TI - Effect of treatment on survival in an animal model of chronic heart failure. PMID- 9860361 TI - Modulation of arterial thrombosis by angiotensin-converting enzyme inhibition and angiotensin II type 1-receptor blockade. PMID- 9860362 TI - Disruption of the endothelial nitric oxide synthase gene: effect on vascular response to injury. PMID- 9860363 TI - Gene therapy for hypertension: antisense inhibition with adeno-associated viral vector delivery targeting angiotensin II type 1-receptor messenger ribonucleic acid. PMID- 9860364 TI - Vascular endothelial growth factor: gene therapy and therapeutic angiogenesis. PMID- 9860365 TI - Changing the treatment paradigm for coronary artery disease. PMID- 9860366 TI - More than coronary artery disease. PMID- 9860367 TI - Need for a paradigm shift: the importance of risk factor reduction therapy in treating patients with cardiovascular disease. AB - Cardiovascular disease remains the number one killer in the United States, despite advances made in diagnosis and therapy. A major shift to expand treatment beyond symptomatic obstructions and infarctions toward comprehensive therapies aimed at treatment of the underlying disease process could decrease the death rate and cost of cardiovascular disease enormously. In the past 5 years, major trials have clearly demonstrated that aggressive intervention with lipid-lowering therapy can dramatically alter the course of disease. Aspirin, smoking cessation, exercise, diet, and other medical and lifestyle interventions can also decrease risk. Successful therapies are not being implemented, however. Making prevention the primary approach to treatment will require increased resource allocation, use of health provider teams, integration of healthcare delivery systems, and expanded emphasis on educating patients about prevention. PMID- 9860368 TI - Why cholesterol as a central theme in coronary artery disease? AB - Evidence from epidemiologic studies and clinical trials have conclusively demonstrated a direct association between coronary artery disease and levels of total and low-density lipoprotein (LDL) cholesterol. Data from a number of studies suggest that even "average" or "normal" cholesterol levels are too high with respect to coronary artery disease risk. Low levels of high-density lipoprotein (HDL) cholesterol have also emerged as a coronary artery disease risk. A recent meta-analysis has eliminated much of the controversy surrounding triglyceride's contribution to coronary artery disease risk, establishing triglyceride levels as an independent risk factor. Lowering lipid levels by any means-including pharmacologic, surgical, and dietary/lifestyle changes--decreases coronary artery disease risk. PMID- 9860369 TI - Diet, lifestyle, and the etiology of coronary artery disease: the Cornell China study. AB - Investigators collected and analyzed mortality data for >50 diseases, including 7 different cancers, from 65 counties and 130 villages in rural mainland China. Blood, urine, food samples, and detailed dietary data were collected from 50 adults in each village and analyzed for a variety of nutritional, viral, hormonal, and toxic chemical factors. In rural China, fat intake was less than half that in the United States, and fiber intake was 3 times higher. Animal protein intake was very low, only about 10% of the US intake. Mean serum total cholesterol was 127 mg/dL in rural China versus 203 mg/dL for adults aged 20-74 years in the United States. Coronary artery disease mortality was 16.7-fold greater for US men and 5.6-fold greater for US women than for their Chinese counterparts. The combined coronary artery disease mortality rates for both genders in rural China were inversely associated with the frequency of intake of green vegetables and plasma erythrocyte monounsaturated fatty acids, but positively associated with a combined index of salt intake plus urinary sodium and plasma apolipoprotein B. These apolipoproteins, in turn, are positively associated with animal protein intake and the frequency of meat intake and inversely associated with plant protein, legume, and light-colored vegetable intake. Rates of other diseases were also correlated with dietary factors. There was no evidence of a threshold beyond which further benefits did not accrue with increasing proportions of plant-based foods in the diet. PMID- 9860370 TI - Atherosclerosis: a nutritional disease of childhood. AB - The development of coronary atherosclerosis begins in childhood. A clear relation between diet and cardiovascular disease risk has been demonstrated. Findings from the Bogalusa Heart Study indicate that most children still exceed national recommendations for intake of total and saturated fat. In addition, children's mean total energy intake is greater than energy expenditure, contributing to the high prevalence of obesity beginning in childhood. Even in childhood, obesity often occurs with other risk factors for cardiovascular disease, such as increased blood pressure, adverse changes in serum lipoproteins, and hyperinsulinemia. This clustering of risk factors has been linked to acceleration of atherosclerotic lesions in the coronary arteries of young individuals. Decreasing the incidence of coronary artery disease in mid and late life necessitates healthy habits in nutrition and lifestyle in early life. Public health measures to favorably alter lifestyle can have a major impact on heart disease prevention and should be pursued vigorously. PMID- 9860371 TI - Determinants of atherosclerosis in the young. Pathobiological Determinants of Atherosclerosis in Youth (PDAY) Research Group. AB - A multicenter cooperative study, Pathobiological Determinants of Atherosclerosis in Youth, examined the relation of the risk factors for adult coronary artery disease to atherosclerosis in nearly 3,000 persons, aged 15-34 years, who died from accidents, homicides, and suicides and were autopsied in forensic laboratories. Very-low-density lipoprotein (VLDL) plus low-density lipoprotein (LDL) cholesterol levels were positively, and high-density lipoprotein (HDL) cholesterol levels were negatively, associated with both fatty streaks and raised lesions in the aorta and right coronary artery, particularly after age 25. Elevated glycohemoglobin levels were associated with raised lesions throughout the 15-34-year age span. Body mass index was associated with both fatty streaks and raised lesions of the right coronary artery in men but not in women. Smoking was associated with a 3-fold increase in raised lesions of the abdominal aorta in the 25-34-year age group. Women lagged after men in the extent of raised lesions in the right coronary artery by about 5 years, but the effects of risk factors in women, except for adiposity, were similar to those in men. The risk factors for adult coronary artery disease accelerate atherogenesis in the teenage years and their effects are amplified in young adulthood, 20-30 years before coronary artery disease becomes clinically manifest. Long-range prevention of adult coronary artery disease will require control of the risk factors early in life. PMID- 9860372 TI - Atherosclerosis and acute coronary events. AB - Without thrombosis, coronary atherosclerosis is generally benign. It is plaque disruption, or fissuring, and subsequent thrombosis that make coronary atherosclerosis dangerous. Small ruptures often remain clinically silent, whereas more extensive plaque rupture may cause the development of unstable angina, myocardial infarction, and sudden death. The risk of plaque disruption depends more on plaque type (composition) than on plaque size and stenosis severity. Both plaque vulnerability (intrinsic disease) and rupture triggers (extrinsic forces) are important for plaque disruption. The resultant thrombotic response, which affects the clinical presentation and outcome, is partly determined by the reactivity of the circulating platelets and the balance between the coagulation and fibrinolytic systems. To prevent and treat life-threatening coronary thrombosis, the medical community should seek new ways to identify and treat dangerous, vulnerable plaques and to optimize antithrombotic treatment. PMID- 9860373 TI - Coronary atherosclerosis: is the process focal or diffuse among patients with symptomatic or fatal myocardial ischemia? AB - Coronary atherosclerosis among patients with symptomatic or fatal myocardial ischemia is a diffuse and extensive process, and it affects all 4 of the major (right, left main, left anterior descending, and left circumflex) epicardial arteries. Histologic evaluation of 5-mm arterial segments reveals involvement of the entire lengths of the epicardial arteries, although the size of plaque may vary from segment to segment. The dominant component of atherosclerotic plaque among patients with symptomatic or fatal myocardial ischemia is fibrous tissue. PMID- 9860374 TI - Is aggressive cholesterol control justified? Review of the post-coronary artery bypass graft trial. AB - The Post-Coronary Artery Bypass Graft (CABG) trial was undertaken to compare the efficacy of aggressive low-density-lipoprotein (LDL) cholesterol lowering (<85 mg/dL) with moderate LDL-cholesterol lowering (130-140 mg/dL) in preventing atherosclerotic progression in saphenous vein grafts. In both the aggressive- and moderate-treatment groups, clinicians titrated lovastatin dosages based on individual patients' LDL-cholesterol levels. Based on angiography performed 4-5 years after enrollment, the rate of disease progression was 31% lower in aggressive-treatment patients than in those who received moderate treatment. Compliance with lovastatin therapy was 85-90%. The results confirm that LDL cholesterol levels should be decreased to <100 mg/dL in patients with coronary artery disease. The challenge is to ensure that at-risk patients receive drug therapy in adequate, individualized doses. PMID- 9860375 TI - Importance of endothelial function in mediating the benefits of lipid-lowering therapy. AB - Trials of lipid lowering by various methods have clearly demonstrated the benefits, clinically and angiographically. Evidence of slowed arterial disease progression and even regression has been convincing but modest, at best. For example, among those treated intensively in the Familial Atherosclerosis Treatment Study (FATS), the mean improvement in proximal stenosis severity was <1% per patient, and only 12% of all lesions showed convincing regression. Despite these modest arterial benefits, the associated reductions in major cardiovascular events have been surprisingly great (24-35% in 3 recent large trials and > or =50% in angiographic trials using combination therapies). The process of plaque disruption helps explain this discrepancy. Disruption can be predicted by a large accumulation of core lipid in the plaque and a high density of lipid-laden macrophages in its thinned fibrous cap. Lesions with these characteristics comprise only 10-20% of the overall lesion population but account for 60-90% of the acute clinical events. Lipid-lowering therapy has beneficial effects on these "high-risk" features of plaque morphology. The composite of data presented here supports the hypothesis that lipid-lowering therapy selectively depletes lipids from this relatively small but dangerous subgroup of fatty lesions, effectively stabilizing them. PMID- 9860376 TI - Coronary artery disease: the Scandinavian Simvastatin Survival Study experience. AB - Although hyperlipidemia is a known risk factor for coronary artery disease, lipid lowering agents were not used widely until recently because evidence was lacking that they could prolong life. In 1987, a large clinical trial, the Scandinavian Simvastatin Survival Study (4S), was designed to test whether such therapy could decrease all-cause mortality in patients with documented coronary artery disease. The prospective, randomized, multicenter trial included 4,444 patients who had had angina pectoris or myocardial infarction (MI), serum total cholesterol of 213 310 mg/dL, and serum triglycerides < or =221 mg/dL. Patients received either simvastatin 20-40 mg/day or placebo and were followed for a median of 5.4 years. Therapy decreased total cholesterol an average of 25%; low-density lipoprotein (LDL) cholesterol, 35%; and triglyceride levels, 10%. Therapy increased high density lipoprotein (HDL) cholesterol levels 8%. Although noncardiac death rates were similar among the groups, the relative risk of mortality (from any cause) was decreased 30%, and the relative risk of coronary mortality was decreased 42% in the simvastatin arm. The mortality risk reductions were profound in patients > or =60 years of age. Treatment also significantly decreased the relative risk of coronary events and the need for bypass surgery or coronary angioplasty. Patients with diabetes also benefited significantly from simvastatin therapy. The reductions in relative risk of major coronary events were achieved irrespective of such baseline risk factors as hypertension and smoking and such medication factors as aspirin, beta-blocker, and calcium-antagonist use. Simvastatin therapy has been shown to be cost-effective, decreasing per-patient hospitalization costs by 31% or $3,872 in 1995 dollars. PMID- 9860377 TI - Preventing coronary artery disease in the West of Scotland: implications for primary prevention. AB - The effectiveness of plasma cholesterol reduction therapy is no longer in question. Recent studies of cholesterol-lowering drug therapy demonstrate significant reductions in risks of mortality and coronary events in patients with existing coronary artery disease. The West of Scotland Coronary Prevention Study (WOSCOPS) demonstrated the therapy's effectiveness as primary prevention in patients without demonstrated coronary artery disease. Now that the efficacy of lipid reduction has been demonstrated conclusively, we face the difficult question of deciding what the treatment goals should be, whom to treat, and how to make those decisions cost effectively. These questions have complex social and ethical components and will lead to considerable discussion about how lipid decreasing drugs will be used. PMID- 9860378 TI - The new pathophysiology of coronary artery disease. AB - Our understanding of coronary artery disease risk and the atherosclerotic process has changed greatly in recent years. For example, it is now known that angiographically apparent coronary artery plaque is not the major cause of myocardial infarction (MI). Rather, it is unstable, soft plaque that cannot be seen angiographically that is prone to rupture and result in infarction. Also important are changes in vascular reactivity resulting from diet. Cholesterol levels by themselves reveal little about a patient's coronary artery disease risk. Most infarctions occur in patients who have normal total cholesterol levels. At-risk patients can be identified using the ratio of total-to-high density lipoprotein (HDL) cholesterol levels. The ratio of triglyceride to HDL cholesterol levels is also important. Simple steps to assess patients' risk in practice are outlined. Primary prevention trials demonstrate that coronary artery disease risk can be lowered dramatically with diet and drug therapy. PMID- 9860379 TI - From heart surgery to prevention. AB - The evolution of the management of coronary artery disease over the last 30 years has been dramatic. Medical therapy, which often proved ineffective, was the mainstay of therapy until the mid-1960s. Then, coronary artery bypass grafting (CABG) revolutionized treatment, and the overall strategy became an invasive one. There was another shift to less invasive therapy with the advent of percutaneous transluminal coronary angioplasty (PTCA). Today, we have come nearly full circle with prevention becoming the cornerstone of management. Preventive intervention programs are evolving. They generally make extensive use of nurses and other nonphysician healthcare professionals. The experience of the preventive cardiology program at the Cleveland Clinic Foundation is reviewed. PMID- 9860380 TI - Avoiding revascularization with lifestyle changes: The Multicenter Lifestyle Demonstration Project. AB - The Multicenter Lifestyle Demonstration Project was designed to determine if comprehensive lifestyle changes can be a direct alternative to revascularization for selected patients without increasing cardiac events. A total of 333 patients completed this demonstration project (194 in the experimental group and 139 in the control group). We found that experimental group patients were able to avoid revascularization for at least 3 years by making comprehensive lifestyle changes at substantially lower cost without increasing cardiac morbidity and mortality. These patients reported reductions in angina comparable with what can be achieved with revascularization. PMID- 9860381 TI - Low-fat diets for children: practicality and safety. AB - Despite our knowledge that atherogenesis begins in childhood and is attributable to dietary fat, pediatricians and dietitians generally frown on low-fat diets for children. It is the responsibility of clinicians to dispel the myths that make this conventional wisdom persist and encourage the adoption of a plant-based, low fat diet. The consumption of adequate calories derived from plant-based foods will provide adequate minerals, protein, and calcium. PMID- 9860382 TI - Low-fat school lunch programs: achieving acceptance. AB - A study was carried out to determine whether hands-on classroom experience with low-fat foods would increase children's acceptance of those foods in the school's lunch program. The 9-month project took place at an elementary school in upstate New York. Half of the classrooms served as the intervention group and received classroom experience with new foods; the other half served as the control group and received no classroom experience. Consumption measurements of 16 new foods, introduced at approximately 2-week intervals, were taken for all students who ate school lunches. Compared with children in the control group, those in the intervention group ate significantly more of the new foods when they were offered in the lunch program. The study demonstrated that experiential learning about food in the classroom is an effective way to gain acceptance of diverse, low-fat foods in the school lunch program. PMID- 9860384 TI - Issues in managing coronary artery disease risk: panel discussions. PMID- 9860383 TI - Coronary risk reduction through intensive community-based lifestyle intervention: the Coronary Health Improvement Project (CHIP) experience. AB - Vigorous cholesterol lowering with diet, drugs, or a combination has been shown to slow, arrest, or even reverse atherosclerosis. Residential lifestyle intervention programs have successfully lowered serum cholesterol levels and other coronary risk factors, but they have the disadvantages of high cost and difficulty with long-term adherence. Community-based risk-reduction programs have the potential to effect change at low cost and improve long-term adherence. To assess the effectiveness of, and to develop a model for, such programs, the community-based Coronary Health Improvement Project (CHIP) was developed in Kalamazoo, Michigan. In the intensive (30-day, 40-hour), hospital-based educational program, participants are encouraged to exercise 30 minutes a day and to embrace a largely unrefined plant-food-centered diet that is high in complex carbohydrates and fiber; very low in fat, animal protein, sugar, and salt; and virtually free of cholesterol. A total of 304 enrollees in the first program were at elevated risk of coronary artery and related diseases: 70% were > or =10% above their ideal weight, 14% had diabetes, 47% had hypertension, and 32% had a history of coronary artery disease. Of the enrollees, 288 "graduated" from the program (123 men, 165 women; mean age was 55+/-11 years). Various markers of disease risk, including serum blood lipids and fasting blood glucose concentrations, were measured before and after the program. At 4 weeks, overall improvements in the participants' laboratory test results, blood pressures, weights, and body mass indexes were highly significant (p <0.001). Triglyceride levels decreased significantly (p <0.05) in participants who had elevated triglyceride levels (>200 mg/dL in men, 200-299 mg/dL in women). PMID- 9860385 TI - Esophageal manometry in clinical practice: the need for evidence-based assessment of clinical efficacy. PMID- 9860386 TI - Hepatitis C and diabetes mellitus: an ongoing controversy. PMID- 9860387 TI - Genetic anticipation in Crohn's disease. PMID- 9860388 TI - Guidelines for the management of Helicobacter pylori infection. Ad Hoc Committee on Practice Parameters of the American College of Gastroenterology. PMID- 9860389 TI - Calcaneal ultrasound bone densitometry in inflammatory bowel disease--a comparison with double x-ray densitometry of the lumbar spine. AB - OBJECTIVE: The aim of this study was to measure ultrasound (US) densitometric parameters [Broadband Ultrasound Attenuation (BUA), Speed of Sound (SOS), and stiffness of the os calcis] in patients with inflammatory bowel disease (IBD) and to compare the results with those obtained with conventional x-ray absorptiometry (DXA) of the lumbar spine. METHODS: Twenty-two patients with Crohn's disease (13 with ileal and nine with ileocolonic disease), 11 patients with ulcerative colitis (eight with left-sided and three with pancolitis), and 18 healthy controls. US densitometry of the right heel and DXA of the lumbar spine were performed within the same day. RESULTS: Compared to controls, IBD patients had significantly lower values with both methods, US and DXA. Forty-nine percent of patients had a lumbar T score below -1. Calcaneal SOS and stiffness of these patients were significantly reduced (p < 0.03 and p < 0.05, respectively). Positive significant correlations were found between lumbar DXA and calcaneal US parameters. Lumbar bone density and calcaneal US stiffness correlated inversely with the lifetime prednisone intake (p < 0.03 andp < 0.05, respectively), but not with age or duration of disease. A cut-off level of 80 dB/MHz for calcaneal BUA predicted axial osteopenia correctly in 74%, but some underestimation of spinal BMD was observed, especially in female patients with Crohn's disease. CONCLUSION: US evaluation of the os calcis gives results similar to those of conventional DXA and therefore may be used for screening IBD patients for axial osteoporosis. Because US does not expose patients to radiation, repeated measurements are possible and may be used to assess short term variations and the effect of treatment of IBD-associated bone disease. PMID- 9860390 TI - Achalasia: a critical review of epidemiological studies. AB - Achalasia is one of the earliest recognized gastroenterological conditions. However, several centuries after it was first described, it remains also among the least understood. One of the main reasons for this is the relative rarity of the disease, which has resulted in limited opportunities to conduct investigative research. Few epidemiological studies have been conducted to date, and their data suggest a worldwide incidence estimated at between 0.03-1.1/10(5)/yr. This review of the literature on the epidemiology of achalasia lends support to the idea that pooling of resources and collaboration at an international level is required, if any significant progress in the cause, treatment, and prevention of the disease is to be made. PMID- 9860391 TI - Prevention of first variceal bleed: an appraisal of current therapies. AB - Primary prophylaxis of esophageal variceal hemorrhage (EVH) is an important issue in the management of patients with portal hypertension. Given the high rates of initial variceal hemorrhage and mortality in patients who have not experienced bleeding from varices, there is an urgent need for some form of primary prophylaxis in all patients with large esophageal varices. The aim of this article is to review the various therapies that have been clinically assessed in randomized controlled trials for their efficacy in prevention of initial EVH. Beta-blockers have been found to be useful in primary prophylaxis of EVH, and the consensus at present is that they should be offered to all patients with portal hypertension who are at high risk for EVH. Nitrates and other newer agents are under evaluation. Surgery is not recommended for primary prophylaxis of EVH. Endoscopic sclerotherapy has not been shown unequivocally to be efficacious, and may even be deleterious, possibly related to an unacceptably high complication rate in this clinical setting. However, it may merit further clinical evaluation in light of recent reports of benefit in certain subgroups of patients with portal hypertension. On the other hand, endoscopic variceal ligation, which has an inherently low complication rate and brings about rapid obliteration of varices, may be a better option for primary prophylaxis of EVH. In the future, preprimary prophylaxis, an attractive concept, may be considered. This would involve intervention with pharmacologic agents even before the development of portal hypertension or esophageal varices. PMID- 9860392 TI - The changing use of esophageal manometry in clinical practice. AB - OBJECTIVE: Clinical practice guidelines now advise against the use of esophageal manometry in the early evaluation of unexplained chest pain. We examined data from patients referred for manometric evaluation over a 10-yr period (1987-1996) to see if clinicians were changing practice patterns and whether manometric diagnoses were affected by the changes. METHODS: Principal indications for the procedure and manometric findings were extracted from a review of 1162 subjects referred to a single clinical laboratory. The tracings were analyzed using a standardized classification method and categorized according to a pathophysiology based scheme. Referral indications and manometric diagnoses were compared for the first and second 5-yr periods of study. RESULTS: Chest pain as a referral indication declined from the first to the second half of the study period (odds ratio, 0.44; p < 0.0001), whereas dysphagia and preoperative evaluations became more common (odds ratio, 1.3; p < 0.05; odds ratio, 13.7; p < 0.0001, respectively). Similarly, hypermotility disorders decreased in frequency (odds ratio, 0.63; p = 0.0001), whereas hypomotility disorders increased (odds ratio, 1.6; p < 0.01). The decrease in hypermotility disorders was solely related to a decrease in nonspecific spastic disorders, including nutcracker esophagus (odds ratio, 0.58; p < 0.0001); the proportion of diagnoses of achalasia and diffuse esophageal spasm remained stable. CONCLUSIONS: These data show that practice patterns are already following current guidelines. They also reflect the disillusionment of clinicians with the poor specificity of manometry in chest pain management, the increasing popularity of antireflux surgery, yet the ongoing observation that nonspecific spastic disorders are closely associated with unexplained chest pain and may have a still-undefined pathogenetic role. PMID- 9860393 TI - HCV and diabetes mellitus: evidence for a negative association. AB - OBJECTIVE: Uncontrolled, retrospective clinical studies have recently claimed that HCV infection could trigger the onset of diabetes mellitus (DM). We sought to verify the association between DM and liver diseases of different etiology, stage, and severity in a prospective study including gender- and age-matched controls. METHODS: Two hundred forty-seven patients with liver cirrhosis (184 men, 116 with an associated hepatocellular carcinoma, 34% in Child-Pugh's class A) were evaluated (group 1). One hundred fifty-seven (63.5) of them were HCV positive, 38 (15.5%) HBV positive, 49 (19.8%) alcohol abusers, and three (1.2%) cryptogenic. Two control groups were also included. The first control group consisted of 138 patients with chronic hepatitis due to HCV infection (73.9%), HBV infection (15.9%), or alcohol abuse (10.2%) (group 2). The second control group included 494 patients with an acute osteoarticular trauma, age- and gender matched with patients in group 1 (group 3). RESULTS: Diabetes mellitus was present in 32.3%, 3.6%, and 9.7% of patients in groups 1, 2, and 3, respectively. When compared with controls (group 3), DM was significantly less frequent in group 2 (p < 0.004) and significantly more frequent in group 1 (p < 0.0001). The prevalence of DM was not different among patients with HCV, HBV infection, or alcohol abuse. In group 3, the prevalence of DM appeared to increase steadily with age. On the contrary, in patients with liver cirrhosis (group 1) DM was detected in about 20-30% of cases in all decades of age. In group 2, diabetics were found only in the 7th and 8th decades of life. At multivariate analysis cirrhosis and age were the only two factors independently associated with DM; odds ratios were 12.5 (95% confidence interval [C.I.], 6.74-20.4) for cirrhosis, and 1.47 for age (95% C.I. 0.39-2.55). CONCLUSIONS: Our findings disprove HCV infection as a trigger factor for DM, which should not be listed among the various extrahepatic manifestations of this viral infection. PMID- 9860394 TI - Anticipation in Crohn's disease may be influenced by gender and ethnicity of the transmitting parent. AB - OBJECTIVE: We sought to examine whether anticipation (an earlier age of onset in succeeding generations) is observed in Crohn's disease (CD) patients within the New York metropolitan area, and whether there are differences in the degree of anticipation with respect to gender and ethnicity of the affected parent. METHODS: Sixty-one parent-child pairs both affected by CD were identified; about half of the pairs were of Ashkenazi Jewish descent. An additional 17 pairs of second-degree relatives with CD were also identified. The intergenerational difference in age at diagnosis (AAD) was used to perform regression analysis and the degree of anticipation among subsets of patients separated on the basis of gender and ethnicity of the transmitting parent was determined. RESULTS: The AAD was consistently (90% of the time) lower in the younger member of the 61 parent child pairs (35.3+/-1.6 yr vs 20.8+/-1.1 yr, p = 0.0001). Furthermore, the degree of anticipation was significantly greater for father-child pairs (20.6+/-3.2 yr) than for mother-child pairs (11.7+/-2.1 yr). However, when the patient population where the parent had an AAD of < 28 was analyzed separately, there was a lack of clear-cut evidence of anticipation in the population as a whole. Only when the population was subdivided by ethnicity was there convincing evidence of anticipation in the Jewish population. CONCLUSION: Ascertainment bias may be responsible for the apparent anticipation observed in the CD population as a whole or in the nonJewish CD subgroup. However, the Jewish CD population displays strong evidence of anticipation even after correction for ascertainment bias. PMID- 9860395 TI - Lower esophageal sphincter relaxation characteristics using a sleeve sensor in clinical manometry. AB - OBJECTIVE: We undertook this study to determine the characteristics of swallow induced lower esophageal sphincter (LES) relaxation in the setting of clinical manometry using a standardized methodology. METHODS: We reviewed 170 manometric recordings performed using a perfused manometric assembly with a sleeve sensor and a computer polygraph. Patients were categorized as patient controls, gastroesophageal reflux disease (GERD), diffuse esophageal spasm (DES), or achalasia. Tracing were semiautomatically analyzed for basal LES pressure, LES pressure during deglutitive relaxation (relaxation LES pressure), duration of LES relaxation, timing of LES relaxation, and the success rate of primary peristalsis. RESULTS: Forty-six patient controls, 93 with GERD, five with DES, and 26 with achalasia were identified. GERD and achalasia patients had lower or higher basal LES pressures than patient controls, respectively. Compared with patient controls, achalasia patients had higher relaxation LES pressures, lower percent LES relaxation, and shorter durations of LES relaxation. The best single measure for distinguishing achalasia was the relaxation LES pressure; using the 95th percentile value of patient controls (12 mm Hg) as the upper limit of normal, its sensitivity and positive predictive value for the diagnosis of achalasia were 92% and 88%, respectively. Coupled with the finding of aperistalsis, a relaxation LES pressure > or = 10 mm Hg achieved 100% sensitivity and positive predictive value among these patients. CONCLUSIONS: Sleeve sensor recording is a practical method for clinical manometry that reliably records LES relaxation characteristics and is amenable to both a standardized manometry protocol and a semiautomated analysis routine. Relaxation LES pressure has a high diagnostic value for achalasia. PMID- 9860396 TI - Measurement of superior mesenteric artery flow by means of Doppler ultrasound in early dumping syndrome. AB - OBJECTIVE: Dumping occurs in about 10% of patients after gastric surgery. It has been suggested that early dumping is associated with an abnormal increase in postprandial splanchnic flow, but data from controlled studies are lacking. Therefore we have studied basal and postprandial superior mesenteric artery (SMA) blood flow in patients with dumping and in two control groups. METHODS: Three groups were studied, one group of patients after gastric surgery with early dumping (n = 6), one surgical control group with patients after gastric surgery without dumping symptoms (n = 7), and a healthy control group without previous gastric surgery (n = 10). Blood glucose and heart rate were measured after dumping provocation by oral ingestion of 50 g glucose. SMA blood flow was measured both basally and 20 min after glucose ingestion. RESULTS: Basal SMA flow was similar in the three groups. After glucose ingestion SMA flow was not significantly different between dumping patients and surgical controls. However, stimulated SMA flow in both groups after gastric surgery combined was significantly (p < 0.05) higher than in healthy controls. CONCLUSION: The systemic symptoms associated with early dumping do not result from increased SMA blood flow per se. After gastric surgery patients have an increased postprandial SMA flow irrespective of the presence of dumping. PMID- 9860397 TI - Physiological role of cholecystokinin in gastroprotection in humans. AB - OBJECTIVE: Cholecystokinin (CCK) is able to protect gastric mucosa against acute injury in experimental animals but little is known about the role of this hormone in maintaining mucosal integrity in humans. This double-blind, placebo controlled study was performed in 16 healthy volunteers. It describes the effects of CCK-8 infused intravenously (i.v.) at physiological doses and endogenous CCK released by intraduodenal (i.d.) oleate on gastric mucosal damage, as brought about by ethanol without or with pretreatment with loxiglumide, a selective CCK-A receptor antagonist. METHODS: CCK-8 was infused i.v. 30 min before and throughout the study or i.d. oleate was instilled through a separate duodenal tube. Thirty minutes after the start of i.v. infusion of CCK or i.d. oleate instillation, 100 ml of 50% ethanol spray was applied to the gastric mucosa using an endoscope. Gastroscopy was performed and mucosal lesions were quantified using modified Lanza score. Gastric biopsies were taken from oxyntic mucosa for histological evaluation and gastric content was aspirated for radioimmunoassay of somatostatin. RESULTS: In placebo-treated subjects ethanol caused endoscopic damage, with an average score of 2.8+/-0.2. Histologically, a widespread disruption of surface epithelium and deep hemorrhagic necrotic lesions were observed. Pretreatment with CCK or i.d. oleate markedly reduced the endoscopic lesion score to 0.7+/-0.1 and 0.3+/-0.1, respectively, and in both cases this reduction was accompanied by a significant rise in plasma CCK. Histologically, surface epithelium was still disrupted but deep necrotic lesions were absent. Gastric content collected before and after CCK or oleate showed a several-fold increase of luminally released somatostatin. CONCLUSIONS: Pretreatment with loxiglumide abolished the protective effects of i.v. CCK-8 and i.d. oleate on mucosal lesions induced by ethanol and prevented the rise in intragastric somatostatin, but failed to affect the increments in plasma CCK. Endogenous CCK plays a physiological role in the maintenance of mucosal integrity. This occurs through activation of CCK-A receptors and is associated with an increased gastric luminal release of somatostatin. PMID- 9860398 TI - Gastric myoelectrical activity in patients with cervical spinal cord injury. AB - OBJECTIVE: Dyspeptic symptoms are common in patients with cervical spinal cord injury (SCI). The supraspinal control of sympathetic innervation to the stomach is interrupted in these patients. Gastric emptying has been reported to be delayed in some patients with cervical SCI. Gastric myoelectrical activity is known to regulate gastric motility and is correlated with gastric emptying. The change in gastric myoelectrical activity after cervical SCI is unknown; our aim was to investigate it. METHODS: The study was performed in 12 cervical SCI patients and 14 healthy controls. Gastric myoelectrical activity was recorded using surface electrogastrography for 30 min in the fasting state and 1 h after a standard test meal. Spectral analysis was performed to compute the following parameters from the electrogastrogram; investigated were the percentage of 2-4 cycles/min (cpm) slow waves, the instability coefficient (IC) of the dominant frequency, the postprandial increment of dominant frequency (deltaF), and its power (deltaP). RESULTS: In both fasting and fed states, regular and stable gastric slow waves were observed in both the control group and patients with cervical SCI. The percentage of normal 2-4 cpm slow waves (preprandial, 80.7+/ 3.6% vs 91.5+/-3.7%, p > 0.05; postprandial, 82.0+/-4.4% vs 87.2+/-4.2%, p > 0.05) and IC (preprandial, 0.19+/-0.04% vs 0.28+/-0.05%; postprandial, 0.24+/ 0.04% vs 0.27+/-0.02%, p > 0.05) were not significantly different between the two groups. The dominant frequency and its power were also similar between the two groups, no matter whether in the fast (frequency, 2.92+/-0.3 vs 2.93+/-0.06 cpm; power, 30.05+/-1.29 vs 29.08+/-1.23 dB, p > 0.05) or fed (frequency, 3.17+/-0.07 vs 3.02+/-0.06 cpm; power, 32.55+/-0.90 vs 32.07+/-1.18 dB,p > 0.05) state. The postprandial response measured by deltaF (0.25+/-0.09 vs 0.09+/-0.07 cpm, p > 0.05) and deltaP (2.52+/-1.10 vs 2.24+/-1.20 dB, p > 0.05) were also similar between the two groups. CONCLUSION: Gastric myoelectrical activity was not altered after cervical SCI. PMID- 9860399 TI - Network of inflammatory cytokines and correlation with disease activity in ulcerative colitis. AB - OBJECTIVE: The inflammatory component of most human inflammatory chronic diseases implicates the production of proinflammatory cytokines. Tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL1beta) seem to play an important role in ulcerative colitis (UC) in relevant experimental models. Moreover, antiTNF therapy seems promising experimentally and clinically. However, these cytokines, and TNFalpha more particularly, are hardly seen in vivo in such patients. The mediators of choice, correlated with disease activities or drug efficacy, remain unclear. To characterize in vivo the network of colonic cytokines in patients with UC, and the contribution of the various cytokines to disease activity we performed this study, using the colonic perfusion method. METHODS: A 20-cm colon length was perfused. Perfusate samples were collected for cytokine determination by enzyme-linked immnoassays. Nineteen perfusions were performed in mild to moderate UC, including two successive perfusions in four patients. Six healthy control patients and four having Crohn's disease (CD) with rectal involvement were studied. Endoscopic score, leukocyte scintigraphy, and systemic markers of inflammation were simultaneously quantified. RESULTS: Large amounts of IL1beta, TNFalpha, IL6, and IL8 were produced in UC patients with a highly significant correlation between TNFalpha, IL1beta and IL8 two by two. Multivariate factorial analysis indicated that IL1beta showed the best correlation with disease activity. Locally produced IL6 was strongly associated with circulating platelet counts. Moreover, production of inflammatory cytokines was associated with similar variations of disease activity in the four patients with two successive perfusions performed. The level of inflammatory cytokines in CD was lower than in UC; TNFalpha, IL1beta, and IL6 were not found in any control patients. CONCLUSION: UC appears to be a chronic inflammatory disease characterized by high production of all four proinflammatory cytokines (IL1beta, TNFalpha, IL6, and IL8). These results suggest that colonic perfusion may be a suitable method to evaluate the local anticytokine properties of new drugs, in correlation with disease activity and systemic markers of inflammation. PMID- 9860400 TI - Ulcerative colitis with skip lesions at the mouth of the appendix: a clinical study. AB - OBJECTIVE: We examined the clinical characteristics of ulcerative colitis patients who demonstrated endoscopically discontinuous lesions at the mouth of the appendix. METHODS: Of patients with initial or recurrent active ulcerative colitis who underwent total colonoscopy during the past 3 yr at Osaka City General Hospital, we selected those who had skip lesions in the mouth of the appendix before treatment, and examined their gender, age, disease type, sites of lesions, inflammatory reaction, severity of disease, effects of treatment, and posttreatment course. RESULTS: Discontinuous lesions at the mouth of the appendix were found in 10 patients, who had the following common clinical features: the major lesion was usually present in the lower part of the large bowel including the rectum, many of the patients had suffered an initial attack only, all patients had mild disease, and many of the patients responded quite satisfactorily to treatment with salicylazosulfapyridine. CONCLUSION: Numerous patients with ulcerative colitis with discontinuous lesions at the mouth of the appendix were observed and their clinical characteristics were examined. Determination of the clinical significance of skip lesions in the appendix will contribute to elucidation of the pathogenesis of ulcerative colitis. PMID- 9860401 TI - Effect of disease localization on the anthropometric and metabolic features of Crohn's disease. AB - OBJECTIVE: We conducted this study to evaluate the effect of disease localization on the anthropometric and metabolic characteristics of inactive Crohn's disease (CD). METHODS: Forty-three patients with biopsy or radiology proven CD (23 men; age, 33.8 yr; range, 18-54 yr) in clinical remission (simplified Crohn's diseases activity index [SCDAI] < 3) and not receiving steroid therapy or nutritional support were enrolled in the study. Patients were separated into three subgroups according to disease localization: ileal (n = 16), ileo-colonic (n = 13), and colonic CD (n = 14). Sixty healthy volunteers (26 men; age, 32.0 yr; range, 18-60 yr), matched for age and height were used as control subjects. Body composition was assessed by both anthropometry and bioimpedance analysis and indirect calorimetry was performed to measure energy expenditure and substrate oxidation rates. RESULTS: CD patients showed a lower body weight than controls, both as a whole (61.5 kg; range, 41.5-74.0 vs 66.9 kg; range, 55.0-86.0 kg; p < 0.001) and separated by disease localization (ileal; p < 0.001; ileo-colonic; p < 0.001; and colonic; p < 0.05 vs controls). Fat-free mass (FFM) did not differ between the groups, whereas fat mass was significantly lower in CD patients than in controls (p < 0.001), with the lowest values in ileal and ileo-colonic patients. Basal metabolic rate by kg of FFM was higher in CD patients than in healthy individuals (p < 0.01). Nonprotein respiratory quotient was significantly lower in CD than in controls as a whole (0.79; range, 0.73-0.84 vs 0.83; range, 0.79-0.89; p < 0.001) or separated by disease localization (p < 0.001 each subgroup vs controls), with a consequent higher lipid oxidation rate. A lower dietary lipid intake was found in patients than in controls (p < 0.01). CONCLUSIONS: Regardless of disease localization, CD patients showed a lower fat mass and a higher utilization of lipids than control subjects. Patients with ileal and ileo-colonic disease showed the greater reduction in body weight, compared with control subjects, suggesting they were at a higher risk of malnutrition, probably as a consequence of the simultaneous occurrence of both malabsorption and decreased energy intake. PMID- 9860403 TI - Pregnancy outcome for women with Crohn's disease: a follow-up study based on linkage between national registries. AB - OBJECTIVE: Crohn's disease, characterized by chronic intestinal inflammation, is sometimes followed by malabsorption, which may interfere with embryogenesis and fetal growth. Therefore we examined birthweight, the frequency of preterm birth, and other reproductive outcomes in the offspring of women with Crohn's disease. METHODS: We used a historical registry-based study, with linkage between the Danish National Registry of Patients and the Danish Medical Birth Registry. Included were 510 newborns to mothers with Crohn's disease and 3018 controls in the study period from 1982 to 1992. RESULTS: The average birthweight of newborns to mothers with Crohn's disease was 185 g, 134 g less than expected for primiparas and multiparas. After adjusting for potential confounders the differences were 142 g (95% confidence interval [CI95%] = 76, 208) and 105 g (CI95% = 37, 173), respectively. The risk of low birthweight was increased in Crohn patients (odds ratio [OR] = 2.4; CI95% = 1.6-3.7), as was the risk of preterm birth (OR = 1.6; CI95% = 1.1-2.3). CONCLUSION: We found a lower birthweight in newborns of patients with Crohn's disease, indicating that Crohn's disease or its treatment may influence fetal growth. PMID- 9860402 TI - Corticosteroid therapy augments gastroduodenal permeability to sucrose. AB - OBJECTIVE: The aim of the present study was to investigate whether corticosteroid therapy alters gastroduodenal mucosal permeability and whether permeability alteration is associated with macroscopic mucosal damage. METHODS: Eight patients taking oral corticosteroid therapy (total prednisone-equivalent dose, 1.5+/-0.1 g; duration, approximately 30 days), nine patients with multiple sclerosis taking high-dose intravenous methyl-prednisolone therapy (total dose, 11.7+/-0.5 g; duration, approximately 9 days), and 20 age- and gender-matched controls were studied. Gastroduodenal permeability was determined using sucrose as a site specific permeability probe. Five-hour urine was collected after ingesting 100 g of sucrose and its urinary excretion rate was measured using high-pressure liquid chromatography. Gastroduodenal endoscopy was performed before steroid therapy to exclude subjects with evidence of macroscopic mucosal lesions. The sucrose test and endoscopy were repeated after completion of corticosteroid therapy. RESULTS: The urinary sucrose excretion rates were similar in the control group and in patient groups before corticosteroid therapy. The median excretion rate of sucrose increased four (one to 28)- and eight (two to 35)-fold, respectively, as compared with pretreatment values in patients taking oral steroid and high-dose intravenous methyl-prednisolone therapy (p < 0.01). Considering all patients together, subjects who received a mean prednisone-equivalent dose of 8.4+/-1.5 g exhibited mucosal lesions, whereas patients who received 3.3+/-1.8 g did not (p = 0.06). The post-therapy increments in sucrose excretion rates were associated with neither the presence of macroscopic lesions nor with the total steroid dose received. CONCLUSIONS: Corticosteroid therapy augments gastroduodenal permeability and high doses are associated with macroscopic mucosal lesions. Steroid-induced permeability increase does not appear to be associated with the presence of macroscopic mucosal lesions. PMID- 9860404 TI - Modulation of intravariceal pressure with pentoxifylline: a possible new approach in the treatment of portal hypertension. AB - OBJECTIVE: In this study the effect of the hemorheological agent pentoxifylline on the pressure of esophageal varices was investigated in portal hypertensive cirrhotic patients. METHODS: Intravariceal pressure was measured endoscopically using the direct puncture technique in 20 patients. Measurements were obtained under baseline conditions and 30 min after double-blind administration of pentoxifylline (1.4 mg/kg BW, n = 10 patients) or an identical volume of NaCl 0.9% solution (n = 10 patients). RESULTS: Under baseline conditions, intravariceal pressure was similar in pentoxifylline and placebo groups (17.3+/ 5.5 mm Hg vs 18.8+/-4.6 mm Hg, respectively; p = N.S.). Placebo administration had no significant effect on intravariceal pressure (18.8+/-4.6 mm Hg vs 18.3+/ 4.1 mm Hg; p = N.S.). In contrast, pentoxifylline caused a highly significant reduction of intravariceal pressure, (from 17.3+/-5.5 mm Hg to 11.4+/-5.9 mm Hg; p = 0.0001), the overall mean reduction being 36.1+/-14.1% mm Hg. CONCLUSIONS: We concluded that pentoxifylline, by reducing blood flow viscosity, caused a significant decrease in variceal pressure in patients suffering from portal hypertension. PMID- 9860405 TI - Small bowel motility in primary biliary cirrhosis. AB - OBJECTIVE: Previous studies have shown small bowel motor activity abnormalities in patients with liver cirrhosis of different etiologies, but motility has not been studied in patients with primary biliary cirrhosis. Our aim was to investigate proximal small bowel motility in these patients. METHODS: Twenty-five female patients presenting clinical, biochemical, serological, and histological findings compatible with primary biliary cirrhosis, 10 female patients with nonalcoholic liver cirrhosis, and 10 normal female controls were studied. Motility of the upper small bowel was measured in the fasted state by means of perfused manometric catheters, connected to external transducers and positioned in the small bowel under fluoroscopy. RESULTS: The average amplitude of contractions was significantly decreased in patients with primary biliary cirrhosis compared with other liver cirrhosis (20.2+/-1.0 vs 32+/-2.9 mm Hg). Also, a significantly increased frequency of cluster of contractions and an increased duration of phase II of the migrating motor complex as seen in liver cirrhosis was observed when compared with normals. CONCLUSION: We conclude that primary biliary cirrhosis patients present motor abnormalities of the small intestine similar to those of patients with liver cirrhosis of other etiologies. In addition, a decrease in the amplitude of small bowel contractions was also found in these patients, suggesting a myogenic involvement. PMID- 9860406 TI - Retinal complications and plasma C5a levels during interferon alpha therapy for chronic hepatitis C. AB - OBJECTIVE: The pathogenesis of retinal complications, such as retinal hemorrhage and cotton-wool spots formation, during interferon (IFN) therapy is unclear. We studied the relationship between the presence of retinal complications and levels of plasma-activated complement 5 (C5a), a known potent intravascular aggregator of granulocytes, during IFN-alpha therapy. METHODS: Forty-five patients with chronic hepatitis C but without diabetes mellitus and hypertension were studied. IFN-alpha was used 10 MU per day for 2 wk and 3 times weekly for an additional 22 wk. In 25 patients with IFN therapy, the optic fundi were examined before therapy began, every 4 wk thereafter, and whenever patients complained of visual symptoms. C5a levels were measured before, and during the 4th, 8th, 12th, and 24th wk, and at any time that a retinal complication was discovered. Twenty patients served as IFN-untreated controls. They had six optic fundi examinations, each 4 wk apart. C5a levels were measured three times, 4 wk apart, in 10 controls. RESULTS: No retinal hemorrhage or cotton-wool spots were detected before IFN-alpha therapy or in any of the controls. However, retinal hemorrhage occurred in six patients (24%) during IFN-alpha therapy. Five of six episodes occurred within the first 8 wk. Only three patients with retinal hemorrhage had visual symptoms. Cotton-wool spots developed in four patients with retinal hemorrhage. Retinal hemorrhage and cotton-wool spots resolved gradually despite continuous administration of IFN-alpha. Before IFN therapy and in controls, all C5a levels were <7 ng/ml. When retinal hemorrhage occurred, C5a was significantly increased (27.3+/-15.6 ng/ml,p < 0.01) relative to levels from the same patients before and after the hemorrhage (5.7+/-1.1 ng/ml), and also relative to levels in IFN-a-treated patients without retinal hemorrhage (5.7+/-1.1 ng/ml) and compared with levels in IFN-untreated controls (5.4+/-0.7 ng/ml). CONCLUSIONS: Retinal hemorrhage or cotton-wool spots often occur during IFN-alpha therapy for chronic hepatitis C. This study suggests that a high C5a level may be an important step in the pathogenesis of retinal capillary infarction, hemorrhage, and cotton-wool spots formation. PMID- 9860407 TI - Interferon-alpha-2B and ribavirin in combination for chronic hepatitis C patients not responding to interferon-alpha alone: an Italian multicenter, randomized, controlled, clinical study. AB - OBJECTIVE: The aim of the study was to assess the efficacy of interferon (IFN) alpha-2b and ribavirin in combination in the treatment of chronic hepatitis C (CHC) patients unresponsive to a previous treatment with IFN-alpha-2b alone. METHODS: We conducted a randomized study in 303 CHC patients. One hundred fifty two patients received subcutaneous administration of recombinant IFN-alpha-2b (3 MU thrice weekly) and ribavirin (1000-1200 mg/daily per os), whereas 151 received IFN-alpha-2b alone (6 MU thrice weekly). Both ribavirin and IFN-alpha-2b were given for 24 wk, regardless of treatment response. Alanine aminotransferase (ALT) levels and HCV RNA titer were checked during the treatment period and for a further 24 wk. RESULTS: Normal ALT levels were observed in 64.5% of the patients treated with IFN-alpha and ribavirin and in 22.6% of the patients treated with IFN-alpha alone. In the group of patients receiving IFN-alpha and ribavirin HCV RNA was not detectable in 40% of patients responders and remained undetectable in 44.2% of sustained responders. In the group of patients receiving IFN-alpha alone HCV RNA was not detectable in 24.2% of patients responders and remained not detectable in 33.3% of sustained responders. CONCLUSIONS: A 24-wk treatment course with IFN-alpha and ribavirin given to patients with a previous lack of response to IFN-alpha alone offers a chance of a sustained biochemical and virological response, at least in a subset of such patients. The role of long term therapy in inducing prolonged remission still remains to be explored. PMID- 9860408 TI - Serum alpha-fetoprotein levels and liver histology in patients with chronic hepatitis C. AB - OBJECTIVE: The clinical and morphological significance of a raised alpha fetoprotein (AFP) level in patients with chronic hepatitis C is undefined. We sought to determine the relation between serum AFP level and liver histology in this population. METHODS: We reviewed the clinical and histological records of 200 consecutively evaluated patients with chronic hepatitis C whose serum AFP levels were recorded. Two groups were studied: group I = 125/200 (62%) patients with normal AFP, < 10 ng/ml; and group II = 75/200 (38%) patients with raised AFP, > 10 ng/ml. The groups were compared according to age, gender, duration of disease, histology, and history of alcohol abuse. RESULTS: There was no significant difference in serum AFP based on age, gender, alcohol consumption, or disease duration. Significant histological differences were observed: cirrhosis was present in 57 (45%) patients in group I versus 51 (68%) in group II (p < 0.001). Hepatocellular carcinoma was more frequent in group II (14/75 [19%]) than in group I (1/125 [1%]) (p < 0.001). Ten of 77 (13%) noncirrhotic patients and 51/108 (47%) cirrhotic patients had a raised AFP (p < 0.002; relative risk, 3.262; confidence interval [C.I.], 1.912-5.564). A derived AFP level of 17.8 ng/ml maximized specificity for predicting histological outcome: one of 76 (1.3%), 29/108 (26.8%), and 14/15 (93.3%) patients were noncirrhotic, cirrhotic, or had HCC, respectively. This derived AFP value is 35% sensitive and 98.6% specific for cirrhosis, with a positive predictive value of 97.7%. CONCLUSION: A serum AFP level >17.8 ng/ml strongly suggests the diagnosis of cirrhosis in a population of patients with chronic hepatitis C. PMID- 9860409 TI - Parenteral antibiotic prophylaxis of bacterial infections does not improve cost efficacy of oral norfloxacin in cirrhotic patients with gastrointestinal bleeding. AB - OBJECTIVE: Selective intestinal decontamination with norfloxacin is useful in the prevention of bacterial infections in cirrhotic patients with gastrointestinal bleeding. However, bleeding cirrhotic patients with ascites, encephalopathy, or shock are at high risk to develop bacterial infections in spite of prophylactic norfloxacin. The aim of this study was to assess whether the addition of intravenous ceftriaxone could improve the efficacy of prophylaxis with norfloxacin in these patients. METHODS: Fifty-six cirrhotic patients with gastrointestinal hemorrhage and ascites, encephalopathy, or shock were randomized into two groups: Group 1 (n = 28) received oral norfloxacin 400 mg/12 h for 7 days, and group 2 (n = 28) received norfloxacin plus intravenous ceftriaxone 2 g daily during the first 3 days of admission. RESULTS: Ten patients were excluded because of community-acquired infection, surgery, or death within the first 24 h. The incidence of bacterial infections during hospitalization was 18.1% in group 1 and 12.5% in group 2 (p = NS). The incidence of severe infections (spontaneous bacterial peritonitis, bacteremia, or pneumonia) was also similar in both groups: 9% in group 1 versus 8.3% in group 2 (p = NS). There were no statistical differences between the two groups with respect to duration of hospitalization or mortality. The cost of antibiotic therapy (including prophylaxis and treatment of infections) was significantly higher in group 2. CONCLUSION: These results suggest that the addition of intravenous ceftriaxone during the first 3 days of hospitalization does not improve the cost-efficacy of oral norfloxacin in the prevention of bacterial infections in cirrhotic patients with gastrointestinal bleeding and high risk of infection. PMID- 9860410 TI - Acute effects of transjugular intrahepatic portosystemic stent-shunt (TIPSS) procedure on renal blood flow and cardiopulmonary hemodynamics in cirrhosis. AB - OBJECTIVE: An acute increase in portal pressure is associated with an immediate reduction in renal blood flow. It has been suggested that this supports the presence of an hepatorenal reflex. In this study, we used TIPSS placement as a model to investigate the effect of an acute reduction in portal pressure on renal blood flow and cardiopulmonary hemodynamic parameters. METHODS: Eleven cirrhotic patients were studied during elective TIPSS placement for variceal hemorrhage (n = 9) or refractory ascites (n = 2). Unilateral renal blood flow (RBF) was measured before and at 5, 15, 30, 45, and 60 min after shunt insertion. Heart rate (HR), mean arterial pressure (MAP), right atrial pressure (RAP), mean pulmonary artery pressure (PAP), pulmonary capillary wedge pressure (PCWP), cardiac output (CO), and systemic vascular resistance (SVR) were also measured before and 30 min after TIPSS placement. RESULTS: Despite significant increases in CO (p = 0.001), RAP (p < 0.001), PAP (p < 0.001), and PCWP (p = 0.001), and a fall in SVR (p = 0.003), no change was observed in RBF, HR, or MAP after TIPSS placement. The fall in the portoatrial pressure gradient correlated only with the rise in CO (p < 0.05) and the drop in SVR (p < 0.05). CONCLUSION: Despite the fall in portal pressure and the systemic hemodynamic changes caused by TIPSS placement, there is no immediate effect on RBF. Any improvement in renal function after TIPSS procedure does not appear to be due to an acute increase in RBF. PMID- 9860411 TI - Predicting hospital mortality in cirrhotic patients: comparison of Child-Pugh and Acute Physiology, Age and Chronic Health Evaluation (APACHE III) scoring systems. AB - OBJECTIVE: The severity of hepatic abnormalities and extent of dysfunction of other organ systems influences prognosis for cirrhosis. The Child-Pugh system has been used to classify cirrhotic patients into good, intermediate, or poor risk categories in evaluation and therapy. Disregard for cardiorespiratory, renal, electrolyte balance, and acid base status limits its predictive accuracy. We evaluated the accuracy of Acute Physiology and Chronic Health Evaluation (APACHE III) to predict short term hospital mortality in patients with liver cirrhosis. METHODS: A total of 282 patients were prospectively enrolled. Child-Pugh and APACHE III scores were recorded on day 1 for each patient. RESULTS: Mean age was 51.7+/-11.3 yr, with 65% men and 35% women; 57% presented with upper GI bleeding, 47% encephalopathy, 9% hepatorenal syndrome, and 7% hepatocellular carcinoma. Sixty-three patients (22%) died. Major causes of death were upper GI bleeding 38%, liver failure 21%, hepatorenal syndrome 19%, hepatocellular carcinoma 4%, and spontaneous bacterial peritonitis 6%. Child-Pugh and APACHE III scores for survivors (8.6+/-2.3 and 58.9+/-35.1) were lower than those for nonsurvivors (10.9+/-2.7 and 87.4+/-30.3) (p < 0.001). Using discriminant analysis, APACHE III correctly identified 75% of cases vs 67% of cases for Child-Pugh (p < 0.05). When information regarding ascites and prothrombin time was added to APACHE III, 81% of cases were correctly classified. CONCLUSION: The APACHE III scoring system is superior to Child-Pugh for prognosticating short term survival of cirrhotic patients. Prognostic accuracy of APACHE III can be enhanced by incorporating information regarding ascites and prothrombin time prolongation. PMID- 9860412 TI - Three months of abstinence from alcohol normalizes energy expenditure and substrate oxidation in alcoholics: a longitudinal study. AB - OBJECTIVE: The aim of this study was to evaluate the energy expenditure, substrate oxidation, and body composition in alcoholics during addiction and after several months of abstinence. METHODS: A total of 32 alcoholics without liver cirrhosis and malabsorption were consecutively recruited. A total of 55 social drinkers, matched for gender and height, were studied as a control group. Anthropometry and bioimpedance analysis were performed to assess body composition, and indirect calorimetry was used to measure basal metabolic rate (BMR) and substrate oxidation. Total abstinence was then achieved in 15 subjects. At 1, 2, 3, and 6 months of abstinence, the metabolic variables and the energy intake were re-examined. RESULTS: At enrollment (T0) alcoholics compared to controls showed a significant decrease in body mass index (22.2+/-2.71 vs 23.6+/ 1.3 kg/m2; p < 0.05), fat mass (14.1+/-4.5 vs 16.7+/-3.3 kg; p < 0.01), an increased BMR normalized by fat-free mass (34.5+/-3.7 vs 32.1+/-2.01 kcal/kg/day; p < 0.01), a lower nonprotein respiratory quotient (npRQ: 0.76+/-0.03 vs 0.83+/ 0.03; p < 0.001), with a consequently higher lipid oxidation (0.08+/-0.02 vs 0.04+/-0.02 g/min; p < 0.01), and a lower carbohydrate oxidation (0.05+/-0.02 vs 0.10+/-0.03 g/min; p < 0.01). Although at 1 and 2 months of abstinence the metabolic parameters had improved, only after 3 months of abstinence did alcoholics show values of body mass index (23.2+/-2.6 kg/ m2), fat mass (17.0+/ 5.34 kg), BMR/fat-free mass (33.1+/-2.78 kcal/kg/day), npRQ (0.82+/-0.02), lipid oxidation (0.05+/-0.03 g/min) and carbohydrate oxidation (0.11+/-0.04 g/min) comparable to those of controls; these values remained constant at 6 months. CONCLUSION: Three months of abstinence from alcohol could represent the minimum time necessary to obtain a normalization of the metabolic variables considered and of the nutritional status for these patients, probably related to a regression of the functional alterations of the microsomal ethanol oxidizing system and of mitochondria secondary to chronic ethanol abuse. PMID- 9860413 TI - Value of MR cholangiography in the diagnosis of obstructive diseases of the biliary tree: a study of 58 cases. AB - OBJECTIVE: The aim of this study was to evaluate magnetic resonance cholangiography (MRC) in the diagnosis of biliary tree obstruction. METHODS: Fifty-eight consecutive patients underwent MRC (GYROSCAN ACS II 1.5 Tesla, TSE T2 axial/coronal-MIP sequences) for clinical and biochemical signs of main bile duct obstruction. MRC images were interpreted by two radiologists and consensus was established according to presence or absence of main bile duct dilation, choledocholithiasis, and malignant or benign stricture. MRC was compared to a final diagnosis established by ultrasound and CT in 19 cases, endoscopic retrograde cholangiopancreatography (ERCP) in 25, intraoperative cholangiography and exploration in 14, and clinical, biochemical, and histological presentation when relevant. Included were single or multiple choledocholithiasis (28, including 11 < or = 3 mm), malignant (10) and benign (12) strictures, and intrahepatic cholestasis (9). RESULTS: Overall, MRC was sensitive (94%) and specific (92%) in detecting main bile duct dilation and choledocholithiasis (86 % and 97 %), but was less sensitive (64%) for small stones < or = 3 mm. Sensitivity for stones > 3 mm was 100%. For benign and malignant strictures, MRC was less sensitive (67% and 80%) but remained specific (98% and 96%). In the detection of normal main bile duct, MRC was highly sensitive (100%) and specific (94%). Diagnostic accuracy ranged from 91% to 98%. CONCLUSION: MRC appears to be specific for choledocholithiasis and sensitive except for small stones. Results for biliary stricture are less satisfactory, but remain specific. Our data confirm that MRC can be useful in the diagnostic workup of main bile duct obstruction. PMID- 9860414 TI - Prospective case-cohort study of intestinal colonization with enterococci that produce extracellular superoxide and the risk for colorectal adenomas or cancer. AB - OBJECTIVE: The aim of this study was to determine whether intestinal colonization with enterococci that produce extracellular superoxide (O2*-), a free radical implicated in the development of colorectal cancer, is associated with these lesions or their precursors. METHODS: A prospective case-cohort study was performed by isolating enterococci from stools of consecutive patients undergoing colonoscopy who had no prior history of colonoscopy or colorectal cancer. A food frequency questionnaire was also administered to control for dietary factors known to affect the risk for these lesions. RESULTS: Among 159 evaluable participants were 77 with no precancerous or cancerous pathology, 61 with adenomas <2 cm, 10 with adenomas > or =2 cm, and 11 with cancer. Regression analyses found no associations for those subjects with adenomas of any size or with cancer and colonization with O2*--producing enterococci, any nutrient, or age. For those patients with large adenomas > or = 2 cm or cancer, however, significant associations were noted for age (OR 1.94 per decade, 95% CI 1.2-3.5), beta-carotene (OR 0.44 per 500 microg/1000 kcal/day, 95% CI 0.2-0.8), vitamin A (OR 3.20 per 500 microg/1000 kcal/day, 95% CI 1.2-8.9), and vitamin E (OR 0.09 per 10 mg/ 1000 kcal/day, 95% CI 0.006-0.9), but not colonization with O2*- producing enterococci. Second stools collected 1 yr later, however, often contained dissimilar enterococcal flora, undermining an important study assumption. CONCLUSIONS: Significant associations were found for those with large adenomas or cancer (but not small adenomas), with age, and with foods enriched for vitamin A, vitamin E, and beta-carotene. An association between colonization with O2*--producing enterococci and colorectal adenomas or cancer, however, could not be ascertained, possibly because intestinal enterococcal flora changes over time, leaving a potentially cohesive hypothesis of colon cancer and risk factors as yet unanswered. PMID- 9860415 TI - Epidemiology of peptic ulcer disease in cirrhotic patients: role of Helicobacter pylori infection. AB - OBJECTIVE: The aim of this study was to investigate the clinical and epidemiological factors associated with the appearance of peptic ulcer in patients with cirrhosis and, in particular, the role of Helicobacter pylori infection. METHODS: A total of 201 of 220 consecutive patients included in a prospective study that aimed to evaluate the effect of dietary intervention on cirrhotic complications and survival underwent upper gastrointestinal endoscopy. At entry, an epidemiological and clinical questionnaire was completed and the presence of peptic ulcer disease or esophageal varices at endoscopy was prospectively collected. Sera were obtained and stored at -70 degrees C until analyzed, being tested afterward for Helicobacter pylori antibodies using a commercial ELISA kit. RESULTS: Eleven of 201 patients had borderline anti Helicobacter pylori IgG titers and were excluded from further analysis. In the remaining 190 patients, point prevalence of peptic ulcer was 10.5% and lifetime prevalence 24.7%. Multivariate analysis selected male sex (OR 2.3; 95%CI 1.09 4.89) and Helicobacter pylori seropositivity (OR: 1.7, 95%CI 1.02-2.81) as the variables independently related to peptic ulcer disease. CONCLUSIONS: Male sex and seropositivity for Helicobacter pylori are the major risk factors for peptic ulcer in cirrhosis. PMID- 9860416 TI - The utility of endoscopy in the management of patients with gastroesophageal reflux symptoms. AB - OBJECTIVE: The utility of endoscopy in the management of patients with symptoms of gastroesophageal reflux disease (GERD) is unclear. The purpose of this prospective study was to assess the impact of endoscopy on the subsequent management of patients with uncomplicated reflux symptoms. METHODS: A total of 742 patients underwent endoscopy for symptoms of GERD. Endoscopists recorded the therapy before endoscopy, the findings of endoscopy, and the treatment recommendations after endoscopy. RESULTS: There was no difference in pre endoscopy therapy or grade of esophagitis in subjects undergoing endoscopy for failed therapy versus GERD symptoms alone. After endoscopy, the most common strategy for patients taking omeprazole was to maintain or increase the dose. For those taking an H2 blocker before endoscopy, the most common outcome was to switch the patient to omeprazole, independent of the grade of esophagitis. CONCLUSIONS: Most patients undergoing endoscopy for symptoms of GERD were switched to omeprazole regardless of the endoscopic findings. No esophageal cancer was identified and the incidence of Barrett's esophagus was low. It appears that endoscopy itself did not change the management of patients receiving H2-blocker therapy. A trial of a proton pump inhibitor before endoscopy should be considered. PMID- 9860417 TI - Antro-pyloric contractile patterns and transpyloric flow after meal ingestion in humans. AB - OBJECTIVE: The present study aimed to evaluate the patterns of antral contractility and pylorus opening and closure in relation to transpyloric flow of a nutrient liquid meal. METHODS: Ultrasound images of the antro-pyloro-duodenal tract were continuously tape-recorded in healthy volunteers for 1 h postprandially and were reviewed twice later by independent observers. Episodes and patterns of pylorus opening and closure, antral-wall contractions, and transpyloric flow were assessed separately and their time relations were identified. RESULTS: Transpyloric flow (forward, 56%+/-13%; retrograde, 19%+/ 11%; and to and fro, 25%+/-14%) occurred essentially during episodes of prolonged pylorus opening not associated with occlusive antral or duodenal proximal contraction, which represented the antro-pyloro-duodenal common chamber. The antro-pyloro-duodenal common chamber lasted on average 36+/-12 s and represented 41%+/-12% of the total observation period. Different patterns of antral contractions were identified according to whether they reached or occluded the terminal antrum and pylorus. Preterminal antral contractions (46%+/-12%) did not propagate beyond the proximal two thirds of the antrum. Terminal antral contractions (54%+/-12%) propagated along the entire antrum and could either occlude (20%+/-9%) or not occlude (42%+/-17%) the lumen. Pyloric opening independent of antral contractions and related to nonocclusive antral contractions could occur equally at the onset of the antro-pyloro-duodenal common chamber. Pyloric closure independent of antral contractions put an end to transpyloric flow in 73%+/-6% of the antro-pyloro-duodenal common chamber. CONCLUSIONS: The final passage of contents from the stomach to the duodenum after the ingestion of a caloric liquid meal is the result of one or more episodes of uni- or bi-directional transpyloric flow, which are regulated by several motor events. The contractile states of the antrum and pylorus show specific features that variably interrelate to provide mixing and to regulate the transpyloric flow of ingesta. A crucial regulator of transpyloric flow appears to be the spatio temporal relation between antral contractions and pyloric closure rather than the contractile events per se. An equal number of pyloric openings related, and unrelated, to antral contractions appears to determine the onset of the antro pyloro-duodenal common chamber and, thus, of transpyloric flow. Pyloric closure unrelated to antral contractions appears to be the main mechanism that interrupts transpyloric flow. In conclusion, it appears that the antro-pyloro-duodenal tract acts as a functional unit in the digestion of a liquid caloric meal. PMID- 9860418 TI - The clinical significance of acquired jejunoileal diverticula. AB - OBJECTIVE: Because of the relative rarity of acquired jejunoileal diverticulosis, including its symptomatology and complications, diagnosis is often difficult and delayed, resulting in unnecessary morbidity and mortality. The purpose of the present study was to draw attention to jejunoileal diverticula and their complications as a site of gastrointestinal symptoms. METHODS: The records of 10 patients with symptomatic jejunoileal diverticula treated in our departments were reviewed. RESULTS: The clinical presentation was varying and nonspecific. Jejunoileal diverticula were diagnosed peroperatively in four patients operated on successfully for their acute complications. In one case the diagnosis was considered after a radiotargeted erythrocyte bleeding scan and in five other cases enteroclysis for chronic abdominal complaints demonstrated jejunoileal diverticula. The death of one patient operated on for massive hemorrhage from jejunal diverticula was probably related to delayed diagnosis and treatment. CONCLUSIONS: Jejunoileal diverticula should not always be dismissed as asymptomatic findings, as they may be the cause of vague, chronic symptomatology and acute complications, including intestinal obstruction, hemorrhage, and perforation. Awareness of the fact that jejunoileal diverticula may cause chronic nonspecific abdominal symptoms and serious acute complications may lead to earlier diagnosis and timely treatment with lower morbidity and mortality. PMID- 9860419 TI - Mycophenolate mofetil in patients with Crohn's disease. AB - OBJECTIVE: Intolerance to azathioprine is a rare but important problem in treating chronically active Crohn's disease. We performed this study to evaluate mycophenolate mofetil as an alternative immunosuppressive therapy for patients with Crohn's disease who did not tolerate azathioprine. METHODS: Four patients with highly active perianal Crohn's disease and two patients with chronically active, steroid-dependent Crohn's disease were included. All patients consumed 2 g/day of mycophenolate mofetil for a median of 8 months (range, 6-12 months). Disease activity was measured by the Perianal Crohn's Disease Activity Index in patients with perianal disease and by the Crohn's Disease Activity Index in patients with chronically active Crohn's disease. RESULTS: Azathioprine-induced side effects disappeared after the drug was discontinued. All patients improved during treatment with mycophenolate mofetil, as shown by a remarkable reduction in the respective clinical scores. Five patients showed no side effects during treatment with mycophenolate mofetil. After 4 months' treatment one patient developed diarrhea that was probably not due to mycophenolate mofetil. CONCLUSION: Mycophenolate mofetil could be an alternative therapy to azathioprine in patients with Crohn's disease. PMID- 9860420 TI - An unusual complication of a biliary stent-small bowel perforation of an incarcerated hernia sac. AB - We present a case of small bowel perforation after migration of an endoscopically inserted biliary stent inside an incarcerated hernia sac. A review of the literature revealed no other report of stent morbidity associated with hernias. The management and implications are discussed. PMID- 9860421 TI - Hormonal therapy in chronic radiation colitis. AB - Severe gastrointestinal bleeding is a rare complication of radiation therapy that requires frequent transfusions. This case report describes a patient with severe bleeding from radiation colitis after treatment of bladder cancer. During 5 months of therapy with multiple drugs, the patient needed 26 units of packed red cells. A subsequent hormone therapy consisting of an estrogen-progesterone combination significantly reduced the need for blood transfusions and hospitalization. We conclude that hormones might provide a promising new additional symptomatic therapy for bleeding radiogenic colitis. PMID- 9860422 TI - A newly recognized entity: intraductal "oncocytic" papillary neoplasm of the pancreas. AB - Intraductal papillary-mucinous tumors of the pancreas are increasingly recognized, and their characteristic endoscopic and radiological features are well reported in the literature in recent years. Oncocytic features in these tumors are uncommon and unrecognized. Intraductal oncocytic papillary neoplasm is a distinct pancreatic tumor and is a recently recognized entity. We report a case of a 69-yr-old patient who presented with symptoms mimicking pancreatitis, resulting in delay in the diagnosis of her pancreatic tumor. She underwent a successful Whipple's procedure and subsequently has remained well. The resected specimen showed an intraductal oncocytic papillary-mucinous neoplasm. The entity is new and the literature information is inadequate at present to judge the biological behavior of this tumor. We discuss this recently recognized entity. PMID- 9860423 TI - Early onset of nephrotic syndrome after treatment with D-penicillamine in a patient with Wilson's disease. AB - Wilson's disease responds to a variety of treatments including D-penicillamine and trientene. Nephrotic syndrome is a late complication of D-penicillamine treatment. We report a pediatric patient with Wilson's disease who developed nephrotic syndrome 2 wk after beginning D-penicillamine. His nephrosis resolved and his disease is quiescent with trientene treatment. PMID- 9860424 TI - Intraduodenal hematoma complicating intestinal biopsy: case reports and review of the literature. AB - We report two cases of postbiopsy duodenal hematoma and review 14 additional cases. Duodenal hematoma predominantly occurs in children and presents with abdominal pain, vomiting, and pancreatitis. Upper gastrointestinal series, abdominal ultrasound, and CT scan are useful in visualizing the hematoma. No comparative studies of the usefulness of these techniques are available, but a CT is indicated if perforation is suspected. The treatment is conservative if no perforation is detected, and resolution of symptoms generally occurs within 2 wk. PMID- 9860425 TI - A case of inflammatory pseudotumor of the liver causing elevated serum CA19-9 levels. AB - Inflammatory pseudotumor of the liver is a rare lesion characterized by proliferating fibrovascular tissue admixed with inflammatory cells. A 50-yr-old Japanese man was hospitalized because of upper abdominal pain and high fever. Computed tomography revealed a poorly demarcated, low density mass in the left lobe of the liver, and abnormal laboratory findings included WBC 9340/mm3, CRP 10.5 mg/dl, and marked elevation of CA19-9 to 1167.9 U/ml. Endoscopic retrograde cholangiography showed irregularity of the intrahepatic bile duct of the left lateral segment, and the lateral segmental branches of the portal vein were not visualized on the venous phase of abdominal angiography. Ultrasound-guided liver biopsy was performed, but malignant disease, including intrahepatic cholangiocarcinoma, could not be completely ruled out. The patient underwent left hepatic lobectomy with lymph node dissection. Histopathological examination yielded a definitive diagnosis of inflammatory pseudotumor. The lesion was immunohistochemically stained for CA19-9 by the ABC method, and the biliary epithelium in severely inflamed portal canals was found to be positive. The markedly elevated preoperative level of CA 19-9 decreased to almost within the normal range and the patient remains well 2 yr 9 months after surgery, without any complications. PMID- 9860426 TI - Arterial thrombosis leading to intestinal infarction in a patient with Behcet's disease associated with protein C deficiency. AB - Behcet's disease may be a possible cause of both occlusive and aneurysmal arterial involvement as well as recurrent venous thrombosis. A case of Behcet's disease complicated with vascular involvement leading to intestinal infarction is presented. A 41-yr-old man suffering from Behcet's disease for 15 yr presented with a 2-day history of severe abdominal pain and bloody diarrhea. Intestinal infarction secondary to thrombosis of the superior mesenteric artery had been diagnosed during surgical exploration 3 yr previously. He was started on anticoagulation with nutritional support. The patient was readmitted with severe diarrhea and malabsorption symptoms 3 yr after intestinal resection. A thrombus located in the posterior wall of the infrarenal portion of aorta was detected by aortography and ultrasonography. Although thrombosis is a relatively common complication of Behcet's disease caused by vasculitis, protein C deficiency, which is a pertinent laboratory finding in this case, might be a secondary factor in the thrombotic event. This is the first case reported of mesenteric artery thrombosis leading to bowel infarction and abdominal aorta thrombosis associated with protein C deficiency. PMID- 9860427 TI - Gastric polypoid lesions--illustrative cases and literature review. AB - Currently, upper gastrointestinal endoscopies are frequently performed for patients with various gastrointestinal symptoms. From time to time, lumps and bumps in the stomach are encountered on endoscopy. Four cases of gastric polypoid lesions are presented. The classification, differentiation, and management approach to these lesions are discussed. Although there is consensus that all gastric adenomatous polyps should be removed, as should gastric hyperplastic polyps that are symptomatic and/or bear dysplastic foci on forceps biopsy, controversy still exists over the management of asymptomatic gastric hyperplastic polyps that do not bear any dysplastic focus on forceps biopsies. Endoscopic ultrasonography (EUS) has a promising role in the evaluation of gastric submucosal polypoid lesions. PMID- 9860428 TI - Acute autoimmune sensorineural hearing loss associated with Crohn's disease. AB - We report the sudden onset of bilateral hearing loss in a patient with Crohn's disease while the intestinal disease was quiescent. Antibodies directed against collagen type II were detected in the serum. Dramatic improvement of his hearing was observed under corticosteroid therapy. Diagnosis of autoimmune sensorineural hearing loss was established. This condition could be considered as an extraintestinal manifestation of Crohn's disease. Early diagnosis is mandated, as prompt and intensive treatment with corticosteroids and/or immunosuppressive drugs is effective and could prevent irreversible hearing loss. PMID- 9860429 TI - A case of progressive multiple focal nodular hyperplasia with alteration of imaging studies. AB - Focal nodular hyperplasia (FNH) of the liver is a lesion characterized by a well circumscribed region of hyperplastic liver tissue with stellate fibrosis. The pathogenesis of the lesion is unknown but various authors consider that FNH may be a response to a preexisting vascular abnormality. We experienced a case of progressive multiple FNH, in which the hemodynamic change as shown by imaging modalities, may support this hypothesis. The patient, a 38-yr-old woman, was found by chance to have multiple portal venous shunts and multiple FNH in both lobes of her liver. Because of their benign characteristics, we followed the nodules periodically without any special treatment. After about 4 yr, the nodules increased both in size and number. In addition, digital subtraction angiography showed that the diameter of the artery had become larger. The hemodynamic change revealed by imaging studies in this case supports the hypothesis that one of the pathogens of FNH is a secondary hepatocellular response to arterial hyperperfusion caused by some vascular malformations. PMID- 9860430 TI - Solitary rectal ulcer induced by excessive use of analgesic suppositories containing paracetamol, caffeine, and codeine. AB - We report the case of a 53-yr-old woman who developed an ulcer of the distal rectum with mild stenosis after prolonged use of suppositories containing paracetamol, caffeine, and codeine. After undergoing extensive diagnostic tests with exclusion of other possible causes, she admitted to the abuse of the suppositories. She was treated with frequent endoscopic balloon dilations to prevent progression of the rectal stenosis. Because of severe pain on defecation, she needed a protective colostomy which could be closed after the healing of the ulcer 7 months later. There was no significant residual stenosis. This case is compared to cases described in the past 30 years. PMID- 9860431 TI - Symptomatic hypocalcemia from oral sodium phosphate: a report of two cases. AB - The use of oral sodium phosphate has, only in very rare instances (seven cases in the literature), been recognized to cause symptomatic hypocalcemia. We report a case in an elderly woman that led to severe tetany and a 10-day admission to control the electrolyte disturbances. The patient was predisposed to developing this complication because of chronic renal failure and vitamin D deficiency. The second hypocalcemic patient had perioral tingling and numbness of the extremities but recovered quickly because of normal renal function. Magnesium depletion was a predisposing factor in the second case. These cases illustrate the rising prevalence of this disorder, which can have severe consequences and can be prevented by avoiding the prescription of these agents to high risk patient groups. PMID- 9860432 TI - Choledochal cysts in adults: a report of two cases and review of the literature. AB - A choledocal cyst is a dilation of some component of the biliary tract that may include both intra- and extra-hepatic sites. They are classified into six types, all of which are relatively rare. Previously, choledochal cysts were treated with biliary-enteric bypass procedures. The current recommendation is to attempt complete excision to minimize the known risk of malignancy and the development of recurrent cholangitis or pancreatitis that may occur in patients with these cystic lesions. Two cases are discussed in which type I choledochal cysts presented. One was removed from a 31-yr-old man who presented with vague abdominal complaints the other from a 32-yr-old man who presented with pancreatitis. The epidemiology, diagnosis, surgical treatment, and risk of cancer in choledochal cysts is described. PMID- 9860433 TI - A diffuse T lymphocytic gastrointestinal mucosal infiltration associated with Sjogren's syndrome resulting in a watery diarrhea syndrome and responsive to immunosuppressive therapy. AB - We report the case of a 45-yr-old white man, investigated for chronic diarrhea, malabsorption and weight loss associated with sicca syndrome. Endoscopic and x ray examinations showed normal macroscopic mucosa in gastrointestinal tract (GIT). Immunohistochemistry showed diffuse polyclonal T cell lymphocytes infiltrating either epithelium and lamina propria in GIT. There was no villous atrophy in the jejunum and ileum. Corticosteroids, azathioprine, and cyclosporine failed to improve symptoms. Monthly intravenous cyclophosphamide administered over 1 yr, stopped the diarrhea and weight loss. The patient is free of symptoms up to a 5-yr follow-up. PMID- 9860434 TI - Granulocytic sarcoma of the jejunum: a rare cause of small bowel obstruction. AB - We report the case of a 40-yr-old man presenting with symptoms of small bowel obstruction. Small bowel x-rays revealed a stricture of the mid-jejunum. Push enteroscopy found a polypoid mass at 1 meter of the ligament of Treitz. Histopathological examination of the biopsy and surgical specimens showed a diffuse infiltrate of the mucosa made of medium to large cells, which were stained on immunohistochemistry by the leucocyte marker CD45 and the histiocyte/monocyte marker CD68 but were negative for the B and T cell markers. Cytological examination of the ascitic fluid revealed many myelobasts with cytoplasmic Auer rods and positive myeloperoxidase staining. There was no evidence of blood or bone marrow involvement suggestive of acute leukemia or myeloproliferative disorders. These findings were consistent with the diagnosis of preleukemic granulocytic sarcoma (or chloroma). Chemotherapy led to complete remission, but 21 months later the patient developed an acute myeloid leukemia. He died from aspergillus pneumonitis, 10 months after bone marrow allograft. Preleukemic granulocytic sarcoma of the small bowel is a rare condition and its diagnosis is usually not easy, requiring histochemical or immunohistochemical studies. Most cases have progressed to acute myeloid leukemia. PMID- 9860435 TI - An unusual hyperplastic hepatocellular nodule in a patient with hepatitis C virus related liver cirrhosis. AB - Recent advances in diagnostic imaging techniques have increased the likelihood of detecting novel nodular lesions of the liver. We report here a case of unusual hyperplastic hepatocellular tumor found in a 70-yr-old woman with hepatitis C virus-related cirrhosis. A mass was incidentally detected in the right lobe by abdominal ultrasonography and confirmed by computed axial tomography (CT). Magnetic resonance imaging demonstrated that the tumor had hyperintense signal with a small hypointense region in the center and a thin, hypointense rim on T1 weighted image and a hypointense signal on T2-weighted image. CT during hepatic arteriography showed that the tumor was hypodense with a central hyperdense region, whereas CT during arterial portography revealed that the tumor was isodense and surrounded by a thin circular hypodense band with a central hypodense region. These radiographic findings suggested a diagnosis of dysplastic nodule with malignant foci of hepatocellular carcinoma. The patient underwent tumor resection. Macroscopically, the tumor, 45 x 45 x 30 mm in size, was encapsulated and had a central stellate-like scar with radiating septa. Histological examination showed a hyperplastic hepatocellular tumor without cellular, nuclear or structural atypia. The central fibrous scar contained abundant small, artery-like and vein-like vessels, whereas there were no normal portal triads but rather several portal tract-like structures lacking bile ducts in the parenchyma of the tumor. Some of the portal tract-like structures were composed of artery-like and vein-like vessels, and the others possessed vein-like vessels only. There were no bile ducts in the tumor. The nontumorous liver tissue had evidence of macronodular cirrhosis. Finally, this tumor was regarded as an unusual type of hyperplastic hepatocellular nodule encountered in cirrhotic liver, characterized by the presence of central stellate-like fibrosis and the lack of bile ducts. Although the pathogenesis of the hyperplastic lesion is unclear, it may represent a focal regenerative hepatocellular response to localized circulatory disorder. PMID- 9860436 TI - Malignant fibrous histiocytoma metastases to the small intestine and colon presenting as an intussusception. AB - A case of malignant fibrous histiocytoma metastases to the small intestine and colon presenting as an intussusception is described. Although malignant fibrous histiocytoma is the most common soft tissue sarcoma in late adult life, GI involvement has rarely been reported. The review of both our case and eight cases in the English-language literature suggests that GI involvement from malignant fibrous histiocytoma occurs most frequently in the small intestine (six of nine) and that two major clinical manifestations of GI involvement are GI bleeding (five of nine) from ulcerated tumors and intussusception (two of nine) led by polypoid tumors. PMID- 9860437 TI - Desiccant recovered, Crohn's disease discovered. AB - Foreign bodies occasionally lodge in strictures due to Crohn's disease. Most cases involve patients known to have Crohn's disease. However, some patients deny or cannot recall ingestion of foreign bodies. Gastrointestinal obstruction caused by a foreign body is a rare presentation of Crohn's disease. We report the first case of Crohn's disease presenting as gastrointestinal obstruction resulting from accidental ingestion of a pharmaceutical desiccant. PMID- 9860438 TI - Common bile duct necrosis after cardiac catheterization. AB - A case of necrosis of the entire extrahepatic biliary tract after cardiac catheterization is presented. The axial blood supply to these structures makes them susceptible to ischemic injury. PMID- 9860439 TI - Asymptomatic elevation of aminotransferase levels and fatty liver secondary to heterozygous hypobetalipoproteinemia. PMID- 9860440 TI - Familial Crohn's disease with systemic lupus erythematosus. AB - We describe a young Japanese woman who was diagnosed with Crohn's disease affecting the ileum, transverse colon, and rectum, as confirmed by barium studies, colonoscopy, and histopathological examination. Her father and sister also had Crohn's disease. After a 4-yr course of sulfasalazine and elemental diet therapy, she was readmitted for perianal abscess associated with the presence of pancytopenia, microhematuria with granular cast, hypocomplementemia, and high titers of autoimmune antibodies (anti-ANA and anti-dsDNA antibodies). Based on these features, a diagnosis of systemic lupus erythematosus (SLE) was made. Despite the rarity of such combination (Crohn's disease with SLE), patients with Crohn's disease who develop such clinical findings might need evaluation for SLE. PMID- 9860441 TI - Acute colitis caused by caustic products. AB - We report two cases of acute proctocolitis caused by rectal application of caustic products of domestic use. One 61-yr-old woman applied an ammonia solution enema; the other patient, a 63-yr-old woman, accidentally applied an enema containing lye. Both patients presented with intense anal pain, but the first patient also had abdominal pain with guarding, hematochezia, and leucocytosis. An acute proctocolitis was found at sigmoidoscopy in both patients. Only conservative and symptomatic measures were prescribed in both cases, and a clinical and endoscopic recovery was seen. In spite of persistent fibrosis in the lamina propria, no signs of stenosis were found. PMID- 9860442 TI - Successful use of an HMG-CoA reductase inhibitor as adjunctive therapy in the management of refractory bile leak complicating laparoscopic cholecystectomy. PMID- 9860443 TI - Klebsiella pneumoniae panophthalmitis: a possible complication of endoscopic variceal injection sclerotherapy. AB - Complication of endoscopic variceal injection sclerotherapy for esophageal variceal hemorrhage is not unusual. However, sclerotherapy complicated panophthalmitis was never reported before. We report such an unusual complication and discuss its possible mechanism and treatment. PMID- 9860444 TI - Pseudo-Mirizzi syndrome in acute cholecystitis. AB - Common hepatic duct obstruction secondary to an impacted cystic duct stone is commonly referred to as the Mirizzi syndrome. Mirizzi syndrome is an uncommon cause of obstructive jaundice and can be mimicked by several other rare conditions. We describe a patient with a massively distended gallbladder due to acute cholecystitis who presented with clinical and cholangiographic findings simulating the Mirizzi syndrome. Endoscopists should be aware of acute cholecystitis as a possible etiology of common hepatic duct obstruction. PMID- 9860445 TI - Fatal intraoperative pulmonary embolism from a hepatic hydatid cyst. AB - A 43-yr-old woman was operated for recurring hydatid cysts of the liver. One of the cysts was located in segment 8 adjacent to both inferior vena cava and right hepatic vein. During the operation, after application of traction on the liver the patient suddenly went into cardiac arrest. After applying open heart massage a Trendelenburg operation was performed, revealing a massive embolus of echinococcal material into the paracentral branches of the pulmonary artery. Resuscitation was unsuccessful. In the literature only four similar cases have been described. The conclusions from these deaths are that an adequate incision is mandatory, no traction on the liver should be necessary, and total vascular exclusion of the liver before cyst drainage and extracorporal bypass are necessary. Interventional techniques should be avoided. PMID- 9860446 TI - Crohn's disease presenting as pyogenic liver abscess with review of previous case reports. AB - A 40-yr-old male doctor from India presented with pyogenic liver abscesses as the first manifestation of Crohn's disease. The Crohn's disease itself was limited to the appendix and the adjacent cecum and could be diagnosed only 6 months after the presentation with liver abscess. This single case highlights three unusual features of Crohn's disease, and stresses the importance of meticulous search for a cause for pyogenic liver abscess when it occurs in an otherwise healthy adult. PMID- 9860447 TI - Accredited fellowship training programs in gastroenterology. PMID- 9860448 TI - The role of vigorous detection of recurrence after curative resection of colorectal cancer. PMID- 9860449 TI - Viper's blood and a bad bug. PMID- 9860450 TI - Healing and prevention of NSAID-associated ulcer disease: is seeing believing? PMID- 9860451 TI - Esophageal dysmotility and its relation to systemic sclerosis variant. PMID- 9860452 TI - Intramuscular approach to liver cholestatic disease. PMID- 9860453 TI - Laparoscopic findings in patients with acute hepatitis by nonhepatitis virus viruses. PMID- 9860454 TI - Has eradication therapy an effect in Helicobacter-positive patients with chronic urticaria? PMID- 9860455 TI - Gastrointestinal involvement in Behcet's syndrome. PMID- 9860457 TI - Water chestnut of the esophagus. PMID- 9860456 TI - Symptoms and Helicobacter pylori: any link? The Italian Helicobacter pylori Study Group. PMID- 9860458 TI - Sherlock Holmes' magnifying glass to identify Wilson's disease? PMID- 9860459 TI - Do fundic gland polyps enter the mainstream of gastric carcinogenesis? PMID- 9860460 TI - Body-fundic mucopeptic cells expansion after Helicobacter pylori eradication. PMID- 9860461 TI - Re: Levine et al. Fecal hydrogen sulfide production in ulcerative colitis. PMID- 9860462 TI - Common bile duct webs. PMID- 9860463 TI - Re: Sharma et al. Dysplasia and cancer risk of short-segment Barrett's esophagus. PMID- 9860464 TI - Fundic gland polyps: a not so innocuous entity worth a careful evaluation. PMID- 9860465 TI - Could serum haptoglobin concentration be used as a marker of hypersplenism in chronic liver disease? PMID- 9860466 TI - Helicobacter pylori infection and gastric juice total phospholipid concentration in patients with bleeding peptic ulcer. PMID- 9860467 TI - Decreased gastroduodenal mucosal concentration of transforming growth factor alpha in Helicobacter pylori-infected dyspeptic patients. PMID- 9860468 TI - Local application of a potent vasoconstrictor during ERCP for the acute management of hemobilia. PMID- 9860469 TI - The European Surgical Association, surgery in Europe, and a taste of the European tomato: founding president address. PMID- 9860470 TI - Cytology of peritoneal lavage performed during staging laparoscopy for gastrointestinal malignancies: is it useful? AB - OBJECTIVE: To evaluate the potential benefit of cytology of the peritoneal lavage obtained during diagnostic laparoscopy for staging gastrointestinal (GI) malignancies. SUMMARY BACKGROUND DATA: Peritoneal lavage is a simple procedure that can be performed during laparotomy for GI tumors. Tumor cells in the lavage fluid are thought to indicate intraperitoneal tumor seeding and to have a negative effect on survival. For this reason, peritoneal lavage is frequently added to diagnostic laparoscopy for staging GI malignancies. METHODS: Patients who underwent peritoneal lavage during laparoscopic staging for GI malignancies between June 1992 and September 1997 were included. Lavage fluids were stained using Giemsa and Papanicolaou methods. Cytology results were correlated with the presence of metastases and tumor ingrowth found during laparoscopy and with survival. RESULTS: Cytology of peritoneal lavage was performed in 449 patients. Tumor cells were found in 28 patients (6%): 8/87 with an esophageal tumor, 2/32 with liver metastases, 11/72 with a proximal bile duct tumor, 7/236 with a periampullary tumor, and none in 7 and 15 patients with a primary liver tumor or pancreatic body or tail tumor, respectively. In 19 of the 28 patients (68%) in whom tumor cells were found, metastatic disease was detected during laparoscopy, and 3 of the 28 patients had a false-positive (n = 1) or a misleading positive (n = 2) lavage result. Therefore, lavage was beneficial in only 6/449 patients (1.3%); in these patients, the lavage result changed the assessment of tumor stage and adequately predicted irresectable disease. Univariate analysis showed a significant survival difference between patients in whom lavage detected tumor cells and those in whom it did not, but multivariate analysis revealed that these survival differences were caused by metastatic or ingrowing disease. CONCLUSION: Cytology of peritoneal lavage with conventional staining should no longer be performed during laparoscopic staging of GI malignancies because it provides an additional benefit in only 1.3% of patients and has limited prognostic value for survival in this group of patients. PMID- 9860471 TI - Laparoscopic cholecystectomy: day-care versus clinical observation. AB - OBJECTIVE: To determine the feasibility and desirability of laparoscopic cholecystectomy (LC) in day-care versus LC with clinical observation. SUMMARY BACKGROUND DATA: Laparoscopic cholecystectomy has been performed regularly as outpatient surgery in patients with uncomplicated gallstone disease in the United States, but this has not been generally accepted in Europe. The main objections are the risk of early severe complications (bleeding) or other reasons for readmission, and the argument that patients might feel safer when observed for one night. Quality-of-life differences hitherto have not been investigated. METHODS: Eighty patients (American Society of Anesthesiology [ASA] I/II) with symptomatic gallstones were randomized to receive LC either in day-care or with clinical observation. Complications, (re)admissions, consultations of general practitioners or the day-care center within 4 days after surgery, use of pain medication, quality of life, convalescence period, time off from professional activities, and treatment preference were assessed. The respective costs of day care and clinical observation were determined. RESULTS: Of the 37 patients assigned to the day-care group who underwent elective surgery, 92% were discharged successfully after an observation period of 5.7+/-0.2 hours. The remainder of the patients in this group were admitted to the hospital and clinically observed for 24 hours. For the 37 patients in the clinical observation group who underwent elective surgery, the observation time after surgery was 31+/ 3 hours. Three patients in the day-care group and one patient in the clinical observation group had complications after surgery. None of the patients in either group consulted a general practitioner or the hospital during the first week after surgery. Use of pain medication was comparable in both groups over the first 48 hours after surgery. There were no differences in pain and other quality of-life indicators between the groups during the 6 weeks of follow-up. Of the patients in the day-care group, 92% preferred day-care to clinical observation. The same percentage of patients in the clinical observation group preferred at least 24 hours of observation to day-care. Costs for the day-care patients were substantially lower (approximately $750/patient) than for the clinical observation patients. CONCLUSION: Effectiveness was equal in both patient groups, and both groups appeared to be satisfied with their treatment. Because no differences were found with respect to the other outcomes, day-care is the preferred treatment in most ASA I and II patients because it is less expensive. PMID- 9860472 TI - Hepatosplanchnic and peripheral tissue oxygenation during treatment of hemorrhagic shock: the effects of pentoxifylline administration. AB - OBJECTIVE: To evaluate the effects of pentoxifylline (PF) administration on liver, gut, and peripheral oxygenation during crystalloid resuscitation of hemorrhagic shock. SUMMARY BACKGROUND DATA: Hypoperfusion of the hepatosplanchnic vascular bed and hypoxia of vital organs may be prolonged despite adequate therapy of hemorrhagic shock. Vasoconstriction, leukostasis, platelet aggregation, and red blood cell plugging could be the underlying causes. PF has been shown to counteract these effects, but its effects in a large animal shock model have been less studied. METHODS: Thirteen anesthetized piglets (mean weight 19.6 kg) were bled steadily to a mean arterial pressure (MAP) of 40 to 50 mmHg and a 70% reduction in cardiac output during 1 hour. These levels were maintained for an additional hour. The animals were resuscitated with acetated Ringer's solution according to MAP and cardiac output values and followed for 80 minutes (total 3 hours and 20 minutes). Seven piglets were given PF boluses (12.5 mg/kg) and infusion (0.2 mg/kg x min), and the rest (n = 6) served as controls. Hemodynamic and systemic oxygen transport variables were recorded. Liver parenchymal and peripheral tissue (subcutaneous, transcutaneous, conjunctival) oxygen tensions (PO2) were measured continuously with polarographic electrodes. Jejunal intramucosal pH (pHi) was calculated every hour by the luminal PCO2, obtained with a balloon tonometer, and arterial bicarbonate concentration. RESULTS: Cardiac output decreased by a mean of 76% during shock and was restored during resuscitation in both groups. MAP decreased from 110 to 40 mmHg but remained at 70 to 80 mmHg during resuscitation in both groups despite remarkable volume load (2.6 ml/min per kg). Liver parenchymal PO2 decreased from 29+/-1 to 15+/-1 mmHg during shock and increased to 36+/-2 mmHg in the PF group, whereas in control group it remained at 26 mmHg. The difference between groups was significant, but at the end of follow-up the liver PO2 decreased to 21 mmHg in both groups. Gut pHi, peripheral tissue oxygen tensions, and the plasma adrenaline and noradrenaline concentrations did not differ between the groups. CONCLUSIONS: Pentoxifylline improved specifically, although only transiently, liver tissue oxygenation. Perhaps the microvascular abnormalities after resuscitation of hemorrhagic shock are more prominent in the hepatic vascular bed, rendering PF specifically effective in that area. The lack of any effect of PF on gut and peripheral tissue oxygenation may have resulted from the persistent vasoconstriction and inadequate restoration of blood volume with crystalloid solution. PMID- 9860473 TI - Inhibition of nitric oxide production and the effects of arginine and Lactobacillus administration in an acute liver injury model. AB - OBJECTIVE: To study the effect of inhibiting nitric oxide production and the effects of arginine and lactobacilli administration in an acute liver injury (LI) model. SUMMARY BACKGROUND DATA: Infectious complications caused by enteric bacteria are common in patients with liver diseases and those who have undergone liver surgery. Increased bacterial translocation has been proposed as one underlying mechanism. Lactobacilli constitute an integral part of the normal gastrointestinal microecology; they are involved in host metabolism and have many beneficial properties. Arginine has numerous roles in cellular metabolism and may be metabolized by lactobacilli in some cases. We have previously shown that rectal administration of Lactobacillus plantarum DSM 9843 (strain 299v), with and without arginine, in an acute LI model significantly reduces the extent of the LI and reduces bacterial translocation. To clarify the pathogenetic mechanisms, we studied the role of nitric oxide in the effects of L. plantarum and arginine in acute LI, as determined by bacterial translocation, ileal, cecal, and colonic nucleotides, RNA, and DNA. METHODS: Male Sprague-Dawley rats were used. L. plantarum, 2% arginine, and/or N-nitro-L-arginine methyl ester (L-NAME), as appropriate, were administered rectally once daily for 8 days. Acute LI was induced on the eighth day by intraperitoneal injection of D-galactosamine (1.1 g/kg body weight), and samples were collected after 24 hours. Bacterial translocation was evaluated by culture of portal and arterial blood, mesenteric lymph nodes, and liver tissue. Liver enzymes and bilirubin were assayed in the serum. The bacterial load in the cecum and colon was determined. Ileal, cecal, and colonic mucosal nucleotides, RNA, and DNA were evaluated. RESULTS: The levels of liver enzymes and bilirubin were lower in liver-injured rats supplemented with arginine and Lactobacillus, and this effect was abolished by the addition of L NAME. Inhibition of nitric oxide production (by L-NAME) increased bacterial translocation in many groups. L-NAME administration increased the cecal and colonic bacterial count and decreased the levels of mucosal nucleotides, RNA, and DNA. CONCLUSIONS: Inhibition of nitric oxide production modulated the effects of arginine and L. plantarum in this acute LI model. L-NAME potentiated the LI, as indicated by elevation of liver enzymes and bilirubin, and it also increased bacterial translocation and the cecal and colonic bacterial count. Increased bacterial translocation could be one of the mechanisms by which LI is potentiated. PMID- 9860475 TI - Isolated hypoxic hepatic perfusion with tumor necrosis factor-alpha, melphalan, and mitomycin C using balloon catheter techniques: a pharmacokinetic study in pigs. AB - OBJECTIVE: To validate the methodology of isolated hypoxic hepatic perfusion (IHHP) using balloon catheter techniques and to gain insight into the distribution of tumor necrosis factor-alpha (TNF), melphalan, and mitomycin C (MMC) through the regional and systemic blood compartments when applying these techniques. SUMMARY BACKGROUND DATA: There is no standard treatment for unresectable liver tumors. Clinical results of isolated limb perfusion with high dose TNF and melphalan for the treatment of melanoma and sarcoma have been promising, and attempts have been made to extrapolate this success to the isolated liver perfusion setting. The magnitude and toxicity of the surgical procedure, however, have limited clinical applicability. METHODS: Pigs underwent IHHP with TNF, melphalan, and MMC using balloon catheters or served as controls, receiving equivalent dosages of these agents intravenously. After a 20-minute perfusion, a washout procedure was performed for 10 minutes, after which isolation was terminated. Throughout the procedure and afterward, blood samples were obtained from the hepatic and systemic blood compartments and concentrations of perfused agents were determined. RESULTS: During perfusion, locoregional plasma drug concentrations were 20- to 40-fold higher than systemic concentrations. Compared with systemic concentrations after intravenous administration, regional concentrations during IHHP were up to 10-fold higher. Regional MMC and melphalan levels steadily declined during perfusion, indicating rapid uptake by the liver tissue; minimal systemic concentrations indicated virtually no leakage to the systemic blood compartment. During isolation, concentrations of TNF in the perfusate declined only slightly, indicating limited uptake by the liver tissue; no leakage of TNF to the systemic circulation was observed. After termination of isolation, systemic TNF levels showed only a minor transient elevation, indicating that the washout procedure at the end of the perfusions was fully effective. CONCLUSIONS: Complete isolation of the hepatic vascular bed can be accomplished when performing IHHP using this balloon catheter technique. Thus, as in extremities, an ideal leakage-free perfusion of the liver can now be performed, and repeated, without major surgery. The effective washout allows the addition of TNF in this setting. PMID- 9860474 TI - Randomized trial of surgery versus surgery followed by adjuvant hepatic arterial infusion with 5-fluorouracil and folinic acid for liver metastases of colorectal cancer. German Cooperative on Liver Metastases (Arbeitsgruppe Lebermetastasen) AB - OBJECTIVE: To determine the impact of adjuvant hepatic arterial infusion (HAI) on survival relative to resection alone in patients with radical resection of colorectal liver metastases. SUMMARY BACKGROUND DATA: Nearly 40% to 50% of all patients with colorectal carcinoma develop liver metastases. Curative resection results in a 5-year survival rate of 25% to 30%. Intrahepatic recurrence occurs after a median of 9 to 12 months in up to 60% of patients. The authors hypothesized that adjuvant intraarterial infusion of 5-fluorouracil (5-FU) might decrease the rate of intrahepatic recurrence and improve survival in patients with radical resection of colorectal liver metastases. METHODS: Between April 5, 1991, and December 31, 1996, patients with colorectal liver metastases from 26 hospitals were stratified by the number of metastases and the site of the primary tumor and randomized to resection of the liver metastases followed by adjuvant HAI of 5-FU (1000 mg/m2 per day for 5 days as a continuous 24-hour infusion) plus folinic acid (200 mg/m2 per day for 5 days as a short infusion), or liver resection only. RESULTS: The first planned intention-to-treat interim analysis after inclusion of 226 patients and 91 events (deaths) showed a median survival of 34.5 months for patients with adjuvant therapy versus 40.8 months for control patients. The median time to progression was 14.2 months for the chemotherapy group versus 13.7 months for the control group. Grade 3 and 4 toxicities (World Health Organization), mainly stomatitis (57.6%) and nausea (55.4%), occurred in 25.6% of cycles and 62.9% of patients. CONCLUSION: According to this planned interim analysis, adjuvant HAI, when used in this dose and schedule in patients with resection of colorectal liver metastases, reduced the risk of death at best by 15%, but at worst the risk of death was doubled. Thus, the chance of detecting an expected 50% improvement in survival by the use of HAI was only 5%. Patient accrual was therefore terminated. PMID- 9860476 TI - Extended drainage versus resection in surgery for chronic pancreatitis: a prospective randomized trial comparing the longitudinal pancreaticojejunostomy combined with local pancreatic head excision with the pylorus-preserving pancreatoduodenectomy. AB - OBJECTIVE: To analyze the efficacy of extended drainage--that is, longitudinal pancreaticojejunostomy combined with local pancreatic head excision (LPJ-LPHE) and pylorus-preserving pancreatoduodenectomy (PPPD) in terms of pain relief, control of complications arising from adjacent organs, and quality of life. SUMMARY BACKGROUND DATA: Based on the hypotheses of pain origin (ductal hypertension and perineural inflammatory infiltration), drainage and resection constitute the main principles of surgery for chronic pancreatitis. METHODS: Sixty-one patients were randomly allocated to either LPJ-LPHE (n = 31) or PPPD (n = 30). The interval between symptoms and surgery ranged from 12 months to 10 years (mean 5.1 years). In addition to routine pancreatic diagnostic workup, a multidimensional psychometric quality-of-life questionnaire and a pain score were used. Endocrine and exocrine functions were assessed in terms of oral glucose tolerance and serum concentrations of insulin, C-peptide, and HbA1c, as well as fecal chymotrypsin and pancreolauryl testing. During a median follow-up of 24 months (range 12 to 36), patients were reassessed in the outpatient clinic. RESULTS: One patient died of cardiovascular failure in the LPJ-LPHE group (3.2%); there were no deaths in the PPPD group. Overall, the rate of in-hospital complications was 19.4% in the LPJ-LPHE group and 53.3% in the PPPD group, including delayed gastric emptying in 9 of 30 patients (30%; p < 0.05). Complications of adjacent organs were definitively resolved in 93.5% in the LPJ LPHE group and in 100% in the PPPD group. The pain score decreased by 94% after LPJ-LPHE and by 95% after PPPD. Global quality of life improved by 71% in the LPJ LPHE group and by 43% in the PPPD group (p < 0.01). CONCLUSIONS: Both procedures are equally effective in terms of pain relief and definitive control of complications affecting adjacent organs, but extended drainage by LPJ-LPHE provides a better quality of life. PMID- 9860477 TI - Moderate activation of the apoptosis inhibitor bcl-xL worsens the prognosis in pancreatic cancer. AB - OBJECTIVE: To analyze the expression of the antiapoptotic gene bcl-xL in human pancreatic cancer and to correlate the results with clinical patient parameters. SUMMARY BACKGROUND DATA: Bcl-xL belongs to the bcl-2-related gene family and acts as a broad antiapoptotic factor to extend both normal and tumor cell survival. Recent findings indicate that tumor cell death induced by chemotherapy and radiotherapy is mediated by the activation of apoptosis. The fact that pancreatic cancer has an extremely malignant potential and that it is resistant to most anticancer treatment modalities suggests that mechanisms are activated that increase the viability of pancreatic cancer cells. METHODS: Seventy-four pancreatic cancer tissue samples were obtained from 32 female and 42 male patients undergoing surgery for exocrine pancreatic cancer. Normal human pancreatic tissue samples were available from 11 organ donors and 4 patients without pancreatic disease. The levels of bcl-xL mRNA expression were analyzed by Northern blot analysis. The exact site of bcl-xL mRNA transcription was determined by nonradioactive in situ hybridization. In addition, immunohistochemistry using specific polyclonal antibodies was used to localize the protein. RESULTS: Northern blot analysis indicated that, in comparison with the normal pancreas, bcl-xL mRNA was markedly overexpressed in 54% of the pancreatic cancer samples. Densitometric analysis revealed that pancreatic adenocarcinomas exhibited a mean 3.4-fold increase (p < 0.01) in bcl-xL mRNA levels in comparison with normal controls. With in situ hybridization, bcl-xL mRNA was found to be highly expressed in the cancer cells of tumor samples that exhibited increased mRNA expression by Northern blot analysis. Immunohistochemical analysis revealed bcl-x immunostaining in 88% of the cancer samples. Correlation of the molecular data with clinical patient parameters revealed that patients whose tumors exhibited no, faint, or weak bcl-xL expression lived significantly longer after tumor resection (median 12 months) than patients whose tumors exhibited moderate bcl-xL mRNA expression (median 5 months) (p < 0.05). However, 5 patients whose tumors exhibited intense bcl-xL mRNA expression tended to live longer (median 14 months). CONCLUSION: Enhanced expression of the antiapoptotic gene bcl-xL in pancreatic cancer and its association with shorter patient survival suggests that this factor may enhance the viability of pancreatic cancer cells in vivo. Inhibition of apoptotic pathways might be one of the reasons why pancreatic cancer shows only limited sensitivity to anticancer treatment. PMID- 9860479 TI - NO release from NO donors and nitrovasodilators: comparisons between oxyhemoglobin and potentiometric assays. AB - Unraveling the biology, pharmacology, and toxicology of NO depends on accurate NO assays, two of the more common being the oxyHb (oxyhemoglobin) assay and potentiometric detection using a Clark-type NO-selective electrode. Comparison of the specificity and sensitivity of the oxyHb and potentiometric methods was carried out using a broad series of nitrovasodilators, including organic nitrates, nitrites, thionitrates, nitrosothiols, and diazenium diolates. Only with the more labile diazenium diolates was a linear relationship observed between the rates of NO release measured potentiometrically and the rate of oxyHb oxidation from the oxyHb assay. The nonlinear plots indicate that N,O-species other than NO itself are capable of oxidizing oxyHb. PMID- 9860478 TI - Warm ischemic tolerance in collapsed pulmonary grafts is limited to 1 hour. AB - OBJECTIVE: To determine the length of warm ischemic tolerance in pulmonary grafts from non-heart-beating donors. SUMMARY BACKGROUND DATA: If lungs could be retrieved for transplant after circulatory arrest, the shortage of donors might be significantly alleviated. Great concern, however, exists about the length of tolerable warm ischemia before cold preservation of pulmonary grafts retrieved from such non-heart-beating donors. METHODS: The authors compared the influence of an increasing postmortem interval on graft function in an isolated, room air ventilated rabbit lung model during blood reperfusion up to 4 hours. Four groups of cadavers (four animals per group) were studied. In group 1, lungs were immediately reperfused. In the other groups, cadavers with lungs deflated were left at room temperature for 1 hour (group 2), 2 hours (group 3), or 4 hours (group 4). RESULTS: Pulmonary vascular resistance was enhanced in all ischemic groups compared with the control group. An increase was noted with longer postmortem intervals in peak airway pressure and in weight gain. A concomitant decline was observed in the venoarterial oxygen pressure gradient caused by progressive edema formation, as reflected by the wet-to-dry weight ratio at the end of reperfusion. CONCLUSIONS: Warm ischemia resulted in increased pulmonary vascular resistance. Graft function in lungs retrieved 1 hour after death was not significantly worse than in nonischemic lungs. Therefore, 60 minutes of warm ischemia with the lung collapsed may be tolerated before cold storage. Further studies are necessary to investigate whether lungs retrieved from non-heart beating donors will become a realistic alternative for transplant. PMID- 9860480 TI - Kinetic study of the reaction of glutathione peroxidase with peroxynitrite. AB - Glutathione peroxidases and their mimics, e.g., ebselen or diaryl tellurides, efficiently reduce peroxynitrite/peroxynitrous acid (ONOO-/ONOOH) to nitrite and protect against oxidation and nitration reactions. Here, we report the second order rate constant for the reaction of the reduced form of glutathione peroxidase (GPx) with peroxynitrite as (8.0 +/- 0.8) x 10(6) M-1 s-1 (per GPx tetramer) at pH 7.4 and 25 degreesC. The rate constant for oxidized GPx is about 10 times lower, (0.7 +/- 0.2) x 10(6) M-1 s-1. On a selenium basis, the rate constant for reduced GPx is similar to that obtained previously for ebselen. The data support the conclusion that GPx can exhibit a biological function by acting as a peroxynitrite reductase. PMID- 9860481 TI - Hypervalent chromium mimics reactive oxygen species as measured by the oxidant sensitive dyes 2',7'-dichlorofluorescin and dihydrorhodamine. AB - Intracellular metabolism of the carcinogen chromate [Cr(VI)] produces the oxidative stress and oxidative DNA damage associated with its genotoxicity. Such oxidative stress has previously been measured by fluorescence using oxidant sensitive dyes and attributed to the formation of reactive oxygen species (ROS). However, metabolism of Cr(VI) also produces Cr(IV) and Cr(V) which can directly damage biological macromolecules without forming ROS. We used the high-valence chromium species, bis(2-ethyl-2-hydroxybutyrato)oxochromate(V) [Cr(V)-EHBA], to test whether high-valence chromium would also react with the oxidant-sensitive dyes 2',7'-dichlorofluorescin (DCFH) and dihydrorhodamine (DHR). Cr(V)-EHBA caused both dyes to fluoresce over a wide dynamic range and under conditions which indicated that Cr(V) had reacted directly with both dyes without first forming a diffusible radical species. Dimethylthiourea (DMTU) and ethanol did not affect Cr(V)-induced fluorescence in vitro or Cr(VI)-induced fluorescence in A549 cells. Under the same conditions, ethanol and DMTU increased the extent of hydrogen peroxide-induced fluorescence. As chromium-induced fluorescence was unaffected by radical scavengers and was qualitatively different from hydrogen peroxide-induced fluorescence, we conclude that DCF and R123 fluorescence in chromate-treated A549 cells is a qualitative and cumulative measure of intracellular Cr(V) formation and not ROS. PMID- 9860482 TI - Additional pathways of S-conjugate formation during interaction of 4 nitrosophenetole with glutathione. AB - The rapid reactions of nitrosoarenes with cellular SH groups have proved to be main metabolic conversions during detoxication. Interactions of the phenacetin metabolite 4-nitrosophenetole with glutathione have been investigated in detail during the last years, revealing a complex pattern of products depending on the stoichiometry of the reactants and reaction conditions. Eight metabolites have been identified hitherto, and the present work extends this medley by six additional products. Three metastable sulfenamides, 4-ethoxy-2,N-bis(glutathion-S yl)-aniline, N4-(glutathion-S-yl)-4-amino-4'-ethoxydiphenylamine, and N (glutathion-S-yl)-4-aminophenol, as well as the N-sulfenylquinonimine N (glutathion-S-yl)-1,4-benzoquinonimine were characterized by chemical reactivity, chromatographic behavior, UV/vis absorption, 1H NMR, and FAB-MS data. The structure of the sulfenamide 2,N4-bis(glutathion-S-yl)-4-amino-4' ethoxydiphenylamine could not be proved unequivocally, but is strongly suggested due to the chemical reactivity, chromatographic behavior, and UV/vis absorption of the compound. Finally, traces of 4-aminophenol were detected. A reaction scheme is presented explaining the formation of all identified metabolites via a central sulfenamide cation. Molecular orbital calculations for this sulfenamide cation have been performed, corroborating the proposed reaction mechanisms on the basis of Klopman's generalized perturbation theory. PMID- 9860483 TI - Formation of 4,4-dialkoxycyclohexa-2,5-dienone N-(thiol-S-yl)imine during reaction of 4-alkoxynitrosobenzenes with thiols in alcoholic solvents. AB - During the interaction of nitrosoarenes with glutathione in aqueous media, intermediate generation of a highly resonance-stabilized sulfenamide cation has been repeatedly suggested. Most intermediates and end products could be explained by reactions of this sulfenamide cation with different nucleophiles such as excess thiol, solvent water, and metabolically produced arylamine. The present paper presents evidence for adduct formation of the sulfenamide cation with solvent alcohol at neutral pH. Sulfenamide cations generated from 4 nitrosophenetole and 4-nitrosoanisole, respectively, are strongly suggested to form the metastable ketals 4-ethoxy-4-methoxycyclohexa-2,5-dienone N-(glutathion S-yl)imine and 4,4-dimethoxycyclohexa-2,5-dienone N-(glutathion-S-yl)imine, respectively, during reaction with solvent methanol. Reaction of the two sulfenamide cations in ethanol yielded 4,4-diethoxycyclohexa-2, 5-dienone N (glutathion-S-yl)imine and 4-ethoxy-4-methoxycyclohexa-2, 5-dienone N-(glutathion S-yl)imine, respectively. Although the metastability of the ketals did not allow isolation of pure solid material, chromatographic and chemical behavior as well as tandem MS fragmentation substantiate a ketal structure of these intermediates. To confirm the proposed structure, new compounds, 2, 6-dimethyl-4 nitrosophenetole, 2,6-dimethyl-4-nitrophenetole, 2, 6-dimethyl-4-phenetidine, and N-(glutathion-S-yl)-N-hydroxy-4-aminoacetophenone, were synthesized and included in supportive experiments. In summary, the detection of ketals corroborates once more the occurrence of a sulfenamide cation which obviously not only reacts with soft nucleophiles such as GSH but, to a limited extent, also reacts with hard nucleophiles. The toxicological significance of this result is discussed. PMID- 9860484 TI - Photoprotective actions of natural and synthetic melanins. AB - Melanins are thought to be important modulators of photochemistry in skin. Eumelanin, a black-brown pigment, is believed to protect against UV-induced photodamage, whereas pheomelanin, a red-yellow pigment, is believed to possess photosensitizing properties. To investigate the hypothesized dichotomy of melanins as both photoprotectants and photosensitizers, we examined the effects of melanins on UV-induced liposomal lipid peroxidation. Sepia melanin, a representative eumelanin, and both red hair pheomelanin and synthetic pheomelanin were employed in these studies. Both eumelanin and pheomelanin inhibited UVA/B- and UVA-induced liposomal lipid peroxidation in a concentration-dependent manner as measured by inhibition of conjugated diene formation. No change in protective properties of the melanins was observed in the presence of saturating levels of O2 during UVA irradiation. Pheomelanin irradiated with UVA/B or UVA induced superoxide-catalyzed reduction of nitroblue tetrazolium, whereas eumelanin did not. Melanins are known to bind various metals, and we examined the effect of iron on the photoproperties of melanins. Eumelanin complexed with Fe(III) did not inhibit UVA/B-induced lipid peroxidation, whereas pheomelanin complexed with Fe(III) stimulated UVA/B-induced lipid peroxidation. Thus, complexation with iron reversed the antioxidant effect of eumelanin and converted pheomelanin into a prooxidant. Analysis of lipid peroxidation products indicated that the oxidation was mediated by free radicals rather than by singlet oxygen. These data indicate that both eumelanin and pheomelanin exert antioxidant effects against UV-induced lipid peroxidation but that the prooxidant activities of pheomelanin result from pheomelanin-metal complexation. PMID- 9860485 TI - Chicken serum albumin hydrolyzes dichlorophenyl phosphoramidates by a mechanism based on transient phosphorylation. AB - The hydrolyzing activities of O-hexyl O-2,5-dichlorophenyl phosphoramidate (HDCP) and p-nitrophenyl butyrate (p-NPB) in chicken serum had been found to copurify in the same protein, identified as albumin. The hydrolyzing activities of both chicken serum and commercial serum albumins from different species were inhibited in a dose-dependent manner by short chain fatty acids. On simultaneous incubation of chicken serum with HDCP and p-NPB, a competitive interaction was detected between the two substrates. This behavior suggests that both are hydrolyzed in the same albumin active site. When chicken serum was preincubated with one of the substrates, and the latter were withdrawn by large dilution, the hydrolyzing activities with both substrates were found to be reduced. This reduction was in turn dependent upon the time of preincubation with the first substrate. These results suggest that HDCP and p-NPB are hydrolyzed by the same albumin active site, via a mechanism based on transient phosphorylation/acylation of the active site. The proposed hydrolysis mechanism would account for the hydrolytic kinetics of both substrates. PMID- 9860486 TI - Prediction of aryl hydrocarbon receptor-mediated enzyme induction of drugs and chemicals by mRNA quantification. AB - Enzyme-specific testing for drug interactions by in vitro techniques has become a routine practice in drug development. With many drugs, enzyme induction has similar importance for the prediction of drug-drug interactions. We developed a method for recognizing enzyme induction mediated via the aryl hydrocarbon receptor. This type of induction may be clinically important since experimental data suggest a higher rate of toxification in induced subjects. Twenty-four drugs and environmental chemicals, selected as prototype inducers or being chemically related to known inducers, including HIV protease inhibitors nelfinavir, saquinavir, ritonavir, and indinavir, were tested for their potency to induce cytochrome P450 1A1 mRNA in human Hela cell cultures by a quantitative reverse transcriptase polymerase chain reaction. Known prototype inducers such as beta naphthoflavone and 3-methylcholanthrene exhibited the highest inducing potency quantified with an Imax value (maximal induction of cytochrome P450 1A1 mRNA synthesis) of 5.48 and 10.7 x 10(6) mRNA molecules per 150 ng of total RNA, respectively. The enzyme-inducing efficacy of some compounds such as resveratrol (2.92 x 10(6)) and the protease inhibitors was not much lower (2.23-3.08 x 10(6)). All compounds that were structurally similar to benzimidazoles exhibited some extent of enzyme induction; e.g., Imax values were 0.86 x 10(6), 0.20 x 10(6), and 0.14 x 10(6) for omeprazole, lansoprazole, and losartan, respectively. To predict the clinical relevance of these inducing effects, the concentration at half-maximal induction IM was estimated; the plasma concentrations of these drug substances were within 1 order of magnitude of the IM values, upon usual dosage. In conclusion, cytochrome P450 1A1 enzyme induction by drugs is a common phenomenon, though there is a great range in the inducing efficacy. In vitro prediction of enzyme induction may be useful for explaining or foreseeing drug interactions, drug side effects, or toxicity by xenobiotics. PMID- 9860487 TI - N2-amination of guanine to 2-hydrazinohypoxanthine, a novel in vivo nucleic acid modification produced by the hepatocarcinogen 2-nitropropane. AB - 2-Nitropropane, an industrial chemical and a hepatocarcinogen in rats, induces aryl sulfotransferase-mediated liver DNA and RNA base modifications [Sodum, R. S., Sohn, O. S., Nie, G., and Fiala, E. S. (1994) Chem. Res. Toxicol. 7, 344 351]. Two of these modifications were previously identified as 8-aminoguanine and 8-oxoguanine. We now report that the base moiety of the so far unidentified third nucleic acid modification, namely RX1 in RNA and DX1 in DNA, is 2 hydrazinohypoxanthine (N2-aminoguanine). 2-Hydrazinoinosine and 2 hydrazinodeoxyinosine, synthesized by adapting published procedures, cochromatographed with RX1 and DX1 of liver RNA and DNA, respectively, from 2 nitropropane-treated rats. 2-Hydrazinoinosine and 2-hydrazinodeoxyinosine are unstable in solution like the in vivo products RX1 and DX1. At neutral pH, hypoxanthine nucleoside is the major product of decomposition, while at pH 10 or above, xanthine nucleoside is also formed. RX1 and DX1 could be generated in the anaerobic reactions of hydroxylamine-O-sulfonic acid, an intermediate in the proposed activation pathway of 2-nitropropane, with guanine nucleosides. These results provide further evidence for the activation of 2-nitropropane and other carcinogenic secondary nitroalkanes to a reactive species capable of aminating nucleic acids and proteins. PMID- 9860488 TI - An unusual DNA adduct derived from the powerfully mutagenic environmental contaminant 3-nitrobenzanthrone. AB - The covalent binding of an N-hydroxy metabolite of the powerfully mutagenic 3 nitrobenzanthrone (NBA) to 2'-deoxyguanosine (dG) and calf thymus DNA has been investigated in vitro. The major adduct obtained from the reaction of the N acetoxy-N-acetyl derivative (N-Aco-N-Ac-ABA) of 3-aminobenzanthrone (ABA) and dG was identified as N-acetyl-3-amino-2-(2'-deoxyguanosin-8-yl)benzanthrone (dG-N-Ac ABA) by 1H NMR and mass spectroscopies as well as by the reaction of N-Aco-N-Ac ABA with the double-stranded calf thymus DNA. The coupling with the dG moiety occurred exclusively at C-2 of benzanthrone (BA), suggesting a significant contribution of a resonance-stabilized arenium ion intermediate derived from BA to the production of this new type of adduct. The preferred conformation of the adduct has been shown to be syn by 1H and 13C NMR. PMID- 9860489 TI - Mutagenic specificity of a derivative of 3-nitrobenzanthrone in the supF shuttle vector plasmids. AB - 3-Nitrobenzanthrone (NBA) is a powerful bacterial mutagen and a suspected human carcinogen present in diesel exhaust and airborne particulates [Enya, T., et al. (1997) Environ. Sci. Technol. 31, 2772-2776]. In the accompanying paper [Enya, T., et al. (1998) Chem. Res. Toxcol. 11, 1460-1467], N-acetoxy-N-acetyl-3 aminobenzanthrone (N-Aco-N-Ac-ABA) was synthesized to yield the DNA adducts of NBA. In this work, to investigate the mutagenic specificity of NBA in human cells, we analyzed mutations induced by N-Aco-N-Ac-ABA using the supF shuttle vector plasmids. Base sequence analysis of 110 and 100 plasmids with mutations in the supF gene propagated in normal cells [WI38-VA13] and nucleotide excision repair deficient cells [XP2OS(SV)], respectively, revealed that the majority of the mutations were base substitutions (85 and 90%) and the rest were deletions and insertions (10 and 15%) in both cell lines. About half of the mutant plasmids had a single base substitution. Of the base substitutions, the most frequent mutation was G.C to T.A transversion (41 and 51%), followed by G.C to A.T transitions (18 and 24%) in either cell. The mutations were distributed not randomly but located at several hot spots, and almost all (nine of ten) hot spots were at the sites of G.C base pairs. The polymerase stop assay in the supF gene revealed that N-Aco-N-Ac-ABA preferentially bound to guanine residues, and mutation sites were generally consistent with the sites where the guanine adducts were formed. PMID- 9860491 TI - Quantitative and qualitative analysis of DNA methylation at N3-adenine by N methyl-N-nitrosourea. AB - The sequence-specific alkylation of DNA by N-methyl-N-nitrosourea (MNU) has been demonstrated for the minor groove N3-methyladenine (N3-MeAde) adduct using neutral thermal hydrolysis and polyacrylamide sequencing gels. The ratio of relative yields of N7- and N3-MeAde and N7-methylguanine (N7-MeGua) is approximately 0.03:0. 15:1.00, respectively, on the basis of the gel data, and these values are comparable to relative yields determined by bulk digestion of MNU-methylated DNA when HPLC was used to analyze the individual adducts. In contrast to the methylation at N7-guanine (N7-Gua) by MNU, alkylation at Ade shows minimal sequence selectivity. Similar to the methylation at N7-Gua, formation of N3-MeAde by MNU is inhibited by 50-200 mM concentrations of NaCl and DNA binding cations, including distamycin and spermine. However, N3-MeAde formation at Ade residues within methidiumpropyl-EDTA-Fe(II) footprinted distamycin DNA affinity binding regions is selectively inhibited at low concentrations of distamycin relative to Ade sites outside of ligand binding regions, and N7-Gua within or outside the distamycin binding regions. HPLC analysis shows that distamycin also quantitatively inhibits the production of N3 methylguanine when calf thymus DNA is treated with MNU or methyl methanesulfonate. The specific inhibitory effect of distamycin, which binds in the minor groove at Ade/Thy-rich sequences, provides additional evidence that the predominant DNA lesion detected at Ade by sequencing gel analysis involves minor groove N3-MeAde modifications. PMID- 9860490 TI - DNA adducts of 2,3-epoxy-4-hydroxynonanal: detection of 7-(1', 2' dihydroxyheptyl)-3H-imidazo[2,1-i]purine and 1,N6-ethenoadenine by gas chromatography/negative ion chemical ionization/mass spectrometry. AB - 2,3-Epoxy-4-hydroxynonanal (EH) is a bifunctional aldehyde formed by epoxidation of trans-4-hydroxy-2-nonenal, a peroxidation product of omega-6 polyunsaturated fatty acids. EH is mutagenic and tumorigenic and capable of modifying DNA bases forming etheno adducts in vitro. Recent studies showed that etheno adducts are present in tissue DNA of humans and untreated rodents, suggesting a potential endogenous role of EH in their formation. A sensitive assay is needed so we can determine whether EH is involved in etheno adduct formation in vivo and study the biological significance of the etheno adducts in DNA. In this study, we developed a gas chromatography/negative ion chemical ionization/mass spectrometry assay for the analysis of 1, N6-ethenoadenine (epsilonAde) and 7-(1', 2'-dihydroxyheptyl) 3H-imidazo[2,1-i]purine (DHH-epsilonAde) in DNA; both are products from the reaction of adenine with EH. The assay entails the following sequence of steps: (1) addition of [15N5]epsilonAde and [15N5]DHH-epsilonAde to DNA as internal standards, (2) acid hydrolysis of DNA, (3) adduct enrichment by C18 solid phase extraction (SPE), (4) derivatization by pentafluorobenzylation (PFB), (5) separation of PFB-epsilonAde and PFB-DHH-epsilonAde on a Si SPE column, (6) acetonide (ACT) formation of PFB-DHH-epsilonAde, and (7) GC/MS analysis with selective ion monitoring (SIM). The limit of detection by on-column injection for PFB-epsilonAde monitoring of the (M - PFB)- ion at m/z 158 was 30 amol and for ACT-PFB-DHH-epsilonAde monitoring of the (M - PFB)- ion at m/z 328 was 0.4 fmol; the detection limits for the entire assay were 6.3 fmol for epsilonAde and 36 fmol for DHH-epsilonAde. In calf thymus DNA modified with EH at 37 degreesC for 50 h, both epsilonAde and DHH-epsilonAde were detected at high levels by this method, 4.5 +/- 0.7 and 90.8 +/- 8.7 adducts/10(3) adenine, respectively. These levels were also verified by HPLC fluorescence analysis, indicating that EH extensively reacts with adenine in DNA, forming etheno adducts. The high sensitivity of the assay suggests that it may be used in the analysis of ethenoadenine adducts in vivo. PMID- 9860492 TI - Stereochemistry of the biotransformation of 1-hexene and 2-methyl-1-hexene with rat liver microsomes and purified P450s of rats and humans. AB - The epoxidation of 1-hexene (1a) and 2-methyl-1-hexene (1b), two hydrocarbons present in the ambient air as pollutants, is catalyzed by some human and rat P450 enzymes. The enantioselectivities of these processes, when the reactions were carried out using rat and human liver microsomal preparations, were modest and dependent on both P450 composition and substrate concentrations. Various P450 isoforms (rat P450 2B1 and human P450 2C10 and 2A6) catalyzed the double bond oxidation of 1a and 1b with different product enantioselectivities. In the case of 1a, a moderately enantioselective hydroxylation at the allylic C(3) with the formation of 1-hexen-3-ol (4a) by microsomes from control or preinduced rats was also observed. The oxidation of this metabolite was, in turn, catalyzed by rat liver microsomes and mainly by rat P450 2C11, leading exclusively to the formation of 1-hexen-3-one, with no double bond epoxidation being observed. The stereochemical course of the microsomal epoxide hydrolase-catalyzed hydrolysis of the epoxy alcohols, threo-(+/-)- and erythro-(+/-)-1, 2-epoxyhexan-3-ol, theoretically expected to be formed from 4a, has been investigated. PMID- 9860493 TI - Mobilization of iron from coal fly ash was dependent upon the particle size and the source of coal. AB - Particulate air pollution, including coal fly ash, contains iron, and some of the pathological effects after inhalation may be due to reactive oxygen species produced by iron-catalyzed reactions. The objective of this study was to determine whether iron, present in coal fly ash, was mobilized, leading to ferritin induction in human airway epithelial cells, and whether the size of the particles affected the amount of iron mobilized. Three types of coal were used to generate the three size fractions of fly ash collected. The Utah coal fly ash was generated from a bituminous b coal, the Illinois coal fly ash from a bituminous c coal, and the North Dakota coal fly ash from a lignite a coal. Three size fractions were studied to compare the amount of iron mobilized in human airway epithelial (A549) cells and by citrate in cell-free suspensions. The size fractions selected were fine (<2.5 microm) and coarse (2.5-10 microm) components of PM10, airborne particulate matter <10 microm in diameter, and the fraction greater than 10 microm. Coal fly ash samples were incubated with 1 mM citrate to determine if iron associated with coal fly ash could be mobilized. Iron was mobilized by citrate from all three size fractions of all three coal types to levels as high as 56.7 nmol of Fe/mg of coal fly ash after 24 h. With all three coal types, more iron was mobilized by citrate from the <2.5 microm fraction than from the >2.5 microm fractions. Further, the mobilized iron was in the Fe(III) form. To determine if iron associated with the coal fly ash could be mobilized by A549 cells, cells were treated with coal fly ash, and the amount of the iron storage protein ferritin was determined after 24 h. Ferritin levels were increased by as much as 11.9-fold in cells treated with coal fly ash. With two of the three types of coal studied, more ferritin was induced in cells treated with the <2.5 microm fraction than with the >2.5 microm fractions. Further, inhibition of the endocytosis of the coal fly ash by the cells resulted in ferritin levels that were near that of the untreated cells, suggesting that iron was mobilized intracellularly, not in the culture medium. The results of this study suggest that differences in particle size and speciation of iron may affect the release of iron in human airway epithelial cells. PMID- 9860495 TI - Hydroxylation and methylthiolation of mono-ortho-substituted polychlorinated biphenyls in rats: identification of metabolites with tissue affinity. AB - The metabolism of three mono-ortho-substituted congeners, 2,3,3',4, 4' pentachlorobiphenyl (CB105), 2,3',4,4',5-pentachlorobiphenyl (CB118), and 2,3,3',4,4',5-hexachlorobiphenyl (CB156), was investigated with regard to the identification of hydroxy- and sulfur-containing metabolites and their tissue retention in rats. Hydroxylation proceeded primarily at the meta or para position either via an arene oxide, involving NIH shift and dechlorination, or by direct insertion of a hydroxyl group. CB105 was hydroxylated preferably in the 2,3,4 trichlorinated ring to yield 4-OH-2,3,3',4', 5-pentaCB, whereas CB118 was hydroxylated in the 2,4, 5-trichlorinated ring to yield the same hydroxy metabolite to a similar extent. The concentration of 4-OH-2,3,3',4',5-pentaCB in blood was >3 times higher than that in liver, lung, or kidney. The ratios of 4-OH 2,3,3',4',5-pentaCB to unchanged CB in blood were 11:1 for CB105 and 7:1 for CB118. The other two metabolites, 4'-OH-2, 3',4,5,5'-pentaCB from CB118 and 4'-OH 2,3,3',4,5,5'-hexaCB from CB156, also exhibited a high blood affinity. Another metabolism of mono-ortho-PCBs PCBs involved methylthiolation in the vicinal ortho and meta unsubstituted positions to give methylthio metabolites, which were detected as methylsulfonyl metabolites in liver and adipose tissue. The tissue retention of these metabolites might contribute to the toxic and biologic effects of mono-ortho-substituted PCBs. PMID- 9860494 TI - Identification of 1,6- and 1,8-dinitropyrene isomers as major mutagens in organic extracts of soil from Osaka, Japan. AB - The organic extracts of soil collected at parks in residential areas in Osaka and neighboring cities in the Kansai area, Japan, showed mutagenicity in Salmonella typhimurium strain TA98 in the presence or absence of a mammalian metabolic activation system (S9 mix). The soil extracts from Ibaraki and two different sites in Osaka, i.e., Sumiyoshi-ku and Minato-ku, were mutagenic in strain TA100 as well as in strain TA98. Direct-acting mutagenicity of soil extracts from Sumiyoshi-ku and Minato-ku toward strain TA98 were 66 or more times higher than that of the other cities. Both extracts exerted stronger mutagenicity in strains YG1021 and YG1024 than TA98 and TA100, and the potency was especially high in strain YG1024: Sumiyoshi-ku, 153 000 revertants/g of soil; and Minato-ku, 246 000 revertants/g of soil. Two mutagenic compounds (I and II) were isolated from the Soxhlet extract of soil from the park in Sumiyoshi-ku by repetitive separation using normal-phase and reversed-phase column chromatography. By comparing the mass and UV spectra and retention times for HPLC on two individual ODS columns of compounds I and II with those of authentic chemicals, we identified these two compounds as 1,6- and 1,8-dinitropyrene (DNPy) isomers. Amounts of DNPy isomers in soil from Sumiyoshi-ku and Minato-ku were 1.7-2.2 ng/g. Forty-three percent and 40% of the mutagenicity of soil from Sumiyoshi-ku and Minato-ku could be attributed to these DNPy isomers, respectively. PMID- 9860496 TI - Identification of free radical formation and F2-isoprostanes in vivo by acute Cr(VI) poisoning. AB - We previously reported the detection of a carbon-centered radical adduct of alpha (4-pyridyl 1-oxide)-N-tert-butylnitrone (POBN) in the bile of rats acutely poisoned with Cr(VI) utilizing an electron spin resonance spin-trapping technique. These former studies suggested that the free radical metabolite was derived from a polyunsaturated fatty acid. The present studies were undertaken to further characterize this radical adduct and to determine whether its formation is associated with enhanced lipid peroxidation in vivo. This report demonstrates that electron spin resonance (ESR) spectra with hyperfine coupling constants aN of 15.71 G and of 2.90 G were present in bile from Cr(VI)-poisoned rats. We found out that virtually identical ESR spectra were obtained when authentic POBN-pentyl radical adducts generated from the reaction of POBN with either pentylhydrazine or linoleic or arachidonic acid with lipoxygenase were added to bile. The hyperfine coupling constants for the POBN-pentyl radical adducts added to bile were as follows: aN = 15.85 G and = 2.60 G for the reaction between pentylhydrazine and POBN; aN = 15.72 G and = 2.61 G for the reaction between arachidonic acid, lipoxygenase, and POBN; and aN = 15.85 G and = 2. 85 G for the reaction between linoleic acid, lipoxygenase, and POBN. In addition, the formation of this radical adduct was associated with lipid peroxidation as quantified by increases in F2-isoprostane levels in bile. These studies, therefore, provide additional evidence that acute Cr(VI) poisoning is associated with enhanced generation of F2-isoprostanes in vivo and tentatively identify the radical species that is produced as the POBN-pentyl radical adduct. PMID- 9860497 TI - Chiral inversion and hydrolysis of thalidomide: mechanisms and catalysis by bases and serum albumin, and chiral stability of teratogenic metabolites. AB - The chiral inversion and hydrolysis of thalidomide and the catalysis by bases and human serum albumin were investigated by using a stereoselective HPLC assay. Chiral inversion was catalyzed by albumin, hydroxyl ions, phosphate, and amino acids. Basic amino acids (Arg and Lys) had a superior potency in catalyzing chiral inversion compared to acid and neutral ones. The chiral inversion of thalidomide is thus subject to specific and general base catalysis, and it is suggested that the ability of HSA to catalyze the reaction is due to the basic groups of the amino acids Arg and Lys and not to a single catalytic site on the macromolecule. The hydrolysis of thalidomide was also base-catalyzed. However, albumin had no effect on hydrolysis, and there was no difference between the catalytic potencies of acidic, neutral, and basic amino acids. This may be explained by different reaction mechanisms of the chiral inversion and hydrolysis of thalidomide. Chiral inversion is deduced to occur by electrophilic substitution involving specific and general base catalysis, whereas hydrolysis is thought to occur by nucleophilic substitution involving specific and general base as well as nucleophilic catalysis. As nucleophilic attack is sensitive to steric properties of the catalyst, steric hindrance might be the reason albumin is not able to catalyze hydrolysis. 1H NMR experiments revealed that the three teratogenic metabolites of thalidomide, in sharp contrast to the drug itself, had complete chiral stability. This leads to the speculation that, were some enantioselectivity to exist in the teratogenicity of thalidomide, it could result from fast hydrolysis to chirally stable teratogenic metabolites. PMID- 9860498 TI - Mechanisms for selective toxicity of fipronil insecticide and its sulfone metabolite and desulfinyl photoproduct. AB - Fipronil, an N-phenylpyrazole with a trifluoromethylsulfinyl substituent, initiated the second generation of insecticides acting at the gamma-aminobutyric acid (GABA) receptor to block the chloride channel. The first generation includes the polychlorocycloalkanes alpha-endosulfan and lindane. In this study, we examine the mechanisms for selective toxicity of the sulfoxide fipronil and its sulfone metabolite and desulfinyl photoproduct relative to their target site interactions in vitro and ex vivo and the importance in fipronil action of biooxidation to the sulfone. Differences in GABA receptor sensitivity, assayed by displacement of 4'-ethynyl-4-n-[2, 3-3H2]propylbicycloorthobenzoate ([3H]EBOB) from the noncompetitive blocker site, appear to be a major factor in fipronil being much more toxic to the insects (housefly and fruit fly) than to the vertebrates (humans, dogs, mice, chickens, quail, and salmon) examined; in insects, the IC50s range from 3 to 12 nM for fipronil and its sulfone and desulfinyl derivatives, while in vertebrates, the IC50 average values are 1103, 175, and 129 nM for fipronil, fipronil sulfone, and desulfinyl fipronil, respectively. The insect relative to the vertebrate specificity decreases in the following order: fipronil > lindane > desulfinyl fipronil > fipronil sulfone > alpha-endosulfan. Ex vivo inhibition of [3H]EBOB binding in mouse brain is similar for fipronil and its sulfone and desulfinyl derivatives at the LD50 dose, but surprisingly, at higher doses fipronil can be lethal without detectably blocking the [3H]EBOB site. The P450 inhibitor piperonyl butoxide, acting in houseflies, increases the metabolic stability and effectiveness of fipronil and the sulfone but not those of the desulfinyl compound, and in mice it completely blocks the sulfoxide to sulfone conversion without altering the poisoning. Thus, the selective toxicity of fipronil and fipronil-derived residues is due in part to the higher potency of the parent compound at the insect versus the mammalian GABA receptor but is also dependent on the relative rates of conversion to the more persistent and less selective sulfone metabolite and desulfinyl photoproduct. PMID- 9860499 TI - N2,7-bis(1-hydroxy-2-oxopropyl)-2'-deoxyguanosine: identical noncyclic adducts with 1,3-dichloropropene epoxides and methylglyoxal. AB - cis- and trans-1,3-dichloropropene epoxides (1,3-D-epoxides) are proposed to be the penultimate or ultimate genotoxic metabolites of the major soil fumigant nematicide 1,3-dichloropropene. The 1, 3-D-epoxide isomers and the potential aldehydes from their degradation readily form adducts with 2'-deoxyguanosine (dGuo) but not with 2'-deoxyadenosine or 2'-deoxycytidine. The reaction of dGuo with the 1,3-D-epoxides (1:20 molar equiv) in phosphate buffer at pH 7.4 for 24 h at 37 degreesC results in complete conversion to four adducts that can be separated by HPLC with the same UV spectra and electrospray (ES)/MS molecular ion and fragmentation patterns. These adducts contain no chlorine and are identical to those obtained more rapidly with methylglyoxal in place of the 1,3-D-epoxides. The four isomeric methylglyoxal adducts with dGuo were proposed originally by others to be the cyclic adducts 1,N2-(1, 2-dihydroxy-2-methyl)ethano-dGuo, but they are reassigned here as the four diastereomers of the noncyclic bis adducts N2, 7-bis(1-hydroxy-2-oxopropyl)-dGuo. The assignments are based on HPLC/UV and HPLC/ES/MS experiments and 1H NMR spectral analysis of the first of the four adducts eluted with HPLC. Acid-catalyzed depurination converts the four dGuo derivatives to the two corresponding isomers of N2,7-bis(1-hydroxy-2 oxopropyl)guanine, assigned by ES/MS and 1H and 13C NMR. Although identical adducts are formed from dGuo with the 1,3-D-epoxides or methylglyoxal, the latter alpha,beta-dicarbonyl compound is not an intermediate in the reaction; instead, the 1,3-D-epoxides hydrolyze to 3-chloro-2-hydroxypropanal which adds to dGuo at N2 and N7. The adducts dehydrochlorinate, in a rate-limiting reaction, thereby giving the same end products obtained on direct reaction with methylglyoxal. Thus, 3-chloro-2-hydroxypropanal (not the 1, 3-D-epoxides or methylglyoxal) is the derivatizing agent for dGuo and therefore probably the mutagenic agent on 1,3 D bioactivation. On the basis of the dGuo model studied here, the DNA adducts of 1, 3-D and its epoxides may be the same as those with methylglyoxal [Vaca, C. E., Fang, J.-L., Conradi, M., and Hou, S.-M. (1994) Carcinogenesis 15, 1887-1894]. PMID- 9860500 TI - Identification of novel metabolites of butadiene monoepoxide in rats and mice. AB - Differences in the metabolism of 1,3-butadiene (Bd) in rats and mice may account for the observed species difference in carcinogenicity. Previous studies of the metabolic fate of Bd have identified epoxide formation as a key metabolic transformation which gives 1, 2-epoxy-3-butene (BMO), although some evidence of aldehyde metabolites is reported. In this study, male Sprague-Dawley rats and male B6C3F1 mice received single doses of [4-14C]BMO at 1, 5, 20, and 50 mg/kg of body weight (0.014, 0.071, 0.286, and 0.714 mmol/kg of body weight). Analysis of urinary metabolites indicated that both species preferentially metabolize BMO by direct reaction with GSH when given by ip administration. The excretion of (R)-2 (N-acetyl-L-cystein-S-yl)-1-hydroxybut-3-ene (IIa), 1-(N-acetyl-L-cystein-S-yl)-2 (S)-hydroxybut-3-ene (IIb), 1-(N-acetyl-L-cystein-S-yl)-2-(R)-hydroxybut-3-ene (IIc), and (S)-2-(N-acetyl-L-cystein-S-yl)-1-hydroxybut-3-ene (IId) accounted for 48-64% of urinary radioactivity in rats and 46-54% in mice. The metabolites originating from the R-stereoisomer of BMO (IIc and IId) predominated over those arising from the S-stereoisomer (IIa and IIb) in both species. IIc was formed preferentially in mice and IId in rats. The corresponding mercaptoacetic acids, S (1-hydroxybut-3-en-2-yl)mercaptoacetic acid (IIf) and S-(2-hydroxybut-3-en-1 yl)mercaptoacetic acid (IIg), were identified only in mouse urine (ca. 20% of the recovered radioactivity). 4-(N-Acetyl-L-cystein-S-yl)-1,2-dihydroxybutane (Ia), a metabolite derived from hydrolysis of BMO, accounted for 10-17% of the radioactivity in rat and 6-10% in mouse urine. 4-(N-Acetyl-L-cystein-S-yl)-2 hydroxybutanoic acid (Ib), 3-(N-acetyl-L-cystein-S-yl)propan-1-ol (Ic), and 3-(N acetyl-L-cystein-S-yl)propanoic acid (Id), also derived from the hydrolysis of BMO, were only present in the rat. Metabolites of 1,2,3,4-diepoxybutane (DEB) were not detected after administration of BMO in rat or mouse urine. This study showed both quantitative and qualitative differences in the metabolism of BMO with varying doses and between species. The data aid in the safety evaluation of Bd and contribute to the interpretation of mathematical models developed for quantitative risk assessment and extrapolation of animals to humans. PMID- 9860501 TI - Probing the mechanism of the carcinogenic activation of N-nitrosodiethanolamine with deuterium isotope effects: in vivo induction of DNA single-strand breaks and related in vitro assays. AB - A series of bioassays, including in vivo induction of DNA single-strand breaks (SSB) and cytotoxicity in cytochrome P450 2E1-transfected cells, were utilized with N-nitrosodiethanolamine (NDELA), its deuterated isotopomers (alpha-D4NDELA and beta-D4NDELA), N-nitroso-2-hydroxymorpholine (NHMOR), and two of its deuterated isotopomers (2-D-NHMOR and 5,5-D2-NHMOR) to probe the mechanism of carcinogenic activation of NDELA and the role of its metabolite NHMOR. DNA samples, taken from the livers of male Wistar rats 4 h after the administration of NDELA, exhibited dose-dependent DNA SSB levels over the range of 0.08-0.75 mmol/kg (body weight), with the greatest SSB level at the highest dose. Deuterium isotope effects on DNA SSB levels were inversely dependent on dose: alpha D4NDELA, 3. 22-1.37; and beta-D4NDELA, 1.38-0.79. At the lowest dose of 0.15 mmol/kg (body weight), 5,5-D2-NHMOR gave an isotope effect for DNA SSB of 2.8 while that for 2-D-NHMOR was 0.7. NDELA and beta-D4NDELA were equally cytotoxic to human P450 2E1-transfected V79 Chinese hamster cells, while alpha-D4NDELA was not. Significant DNA SSB levels were observed in these cells for NDELA and beta D4NDELA but not for alpha-D4NDELA. A kinetic deuterium isotope effect of 2.6 for Vmax/Km was observed for the horse liver alcohol dehydrogenase-mediated oxidation of beta-D4NDELA to NHMOR, while kH/kD for alpha-D4NDELA was 1.05. These data provide the first definitive evidence for the activation of NDELA by a pathway involving the scission of the alpha-CH bond and are consistent with P450 2E1 mediated alpha-hydroxylation of NDELA producing the corresponding reactive alpha hydroxynitrosamine. PMID- 9860502 TI - Lactols in hydrolysates of DNA treated with alpha-acetoxy-N-nitrosopyrrolidine or crotonaldehyde. AB - alpha-Acetoxy-N-nitrosopyrrolidine (alpha-acetoxyNPYR) is a stable precursor to alpha-hydroxyNPYR, the initial product of metabolism and proposed proximate carcinogen of N-nitrosopyrrolidine (NPYR). Crotonaldehyde (2-butenal) is a metabolite of NPYR and also a mutagen and carcinogen. Both alpha-acetoxyNPYR and crotonaldehyde form DNA adducts, but these reactions have not been completely characterized. In previous studies, we detected substantial amounts of unidentified radioactivity in hydrolysates of DNA that had been treated with radiolabeled alpha-acetoxyNPYR. In this study, we have characterized these products as 2-hydroxytetrahydrofuran, the cyclic form of 4-hydroxybutanal, and paraldol, the dimer of 3-hydroxybutanal. These products were identified by comparison to standards and by conversion to 2,4-dinitrophenylhydrazones. 2 Hydroxytetrahydrofuran is the major product in neutral thermal hydrolysates of alpha-acetoxyNPYR-treated DNA and is derived predominantly from N2 (tetrahydrofuran-2-yl)deoxyguanosine 8. Paraldol is present to a lesser extent than 2-hydroxytetrahydrofuran in these reactions and is formed from paraldol releasing adducts, which in turn are produced in the reaction of crotonaldehyde, a solvolysis product of alpha-acetoxyNPYR, with DNA. Other products in hydrolysates of alpha-acetoxyNPYR-treated DNA are N7-substituted guanines 5 and 6, cyclic N7-C8 guanines 4, 11, and 12, and 1, N2-propanodeoxyguanosines 9 and 10. Paraldol is a major product in hydrolysates of crotonaldehyde-treated DNA, being present in amounts 100 times greater than those of previously identified adducts 9 and 10. The results of this study provide a more complete picture of the reactions of alpha-acetoxyNPYR with DNA and yield some new insights about possible endogenous DNA adducts formed from crotonaldehyde. PMID- 9860503 TI - Inhibition of nitrous acid-dependent tyrosine nitration and DNA base deamination by flavonoids and other phenolic compounds. AB - Exposure of tyrosine or DNA bases to acidic nitrite at low pH results in the nitration of tyrosine and the formation of base deamination products, respectively. At pH 1, hypoxanthine and xanthine are formed from the deamination of adenine and guanine, respectively, whereas under the same conditions, uracil is not detected. The yield of 3-nitrotyrosine derived from interaction of equimolar nitrite and tyrosine at pH 1 is approximately 50% of that obtained from equimolar peroxynitrite-tyrosine interactions at pH 7. 4. The ability of a range of plant phenolic constituents to prevent damage mediated by acidic nitrite was also examined in comparison with the activity of vitamin C. The epicatechin/gallate family of flavonols, constituents of green tea, red wine, etc., demonstrates the most extensive inhibitory properties against both tyrosine nitration and base deamination. The results also show that ascorbic acid is a poor inhibitor of nitration or deamination under acidic conditions such as those of the stomach. The ability of plant phenolics to scavenge reactive nitrogen species derived from acidic nitrite may contribute to the protective effects of tea polyphenols against gastric cancer. PMID- 9860504 TI - Action of bleomycin on structural mimics of intermediates in DNA double-strand cleavage. AB - Bleomycin-induced cleavage was examined in several nicked, gapped, or intact duplex DNA substrates, including a structure designed to mimic a proposed singly nicked intermediate in double-strand cleavage. This nicked structure appeared to correctly target the second cleavage event in the complementary strand, resulting in a blunt-ended double-strand break, similar to that induced directly by bleomycin alone in an intact duplex of the same sequence. A one-base-gapped structure was markedly less efficient in correctly targeting bleomycin attack in the complementary strand. The results are consistent with a model of bleomycin induced double-strand cleavage in which the nick formed by the initial bleomycin attack serves to target secondary attack to a specific position in the complementary strand, resulting in a double-strand break with a defined geometry. PMID- 9860505 TI - Metabolism-dependent neutrophil cytotoxicity of amodiaquine: A comparison with pyronaridine and related antimalarial drugs. AB - Life-threatening agranulocytosis and hepatotoxicity during prophylactic administration of amodiaquine have led to its withdrawal. Agranulocytosis is thought to involve bioactivation to a protein-reactive quinoneimine metabolite. The toxicity of amodiaquine and the lack of cheap drugs have prompted a search for alternative antimalarial agents. The aim of this study was to determine the metabolism and neutrophil toxicity of amodiaquine, pyronaridine, and other related antimalarial agents. Horseradish peroxidase and hydrogen peroxide were used to activate drugs to their respective quinoneimine metabolites. Metabolites were trapped as stable glutathione conjugates, prior to analysis by LC/MS. Amodiaquine was metabolized to a polar metabolite (m/z 661), identified as a glutathione adduct. Tebuquine was converted to two polar metabolites. The principal metabolite (m/z 686) was derived from glutathione conjugation and side chain elimination, while the minor metabolite gave a protonated molecule (m/z 496). Only parent ions were identified when chloroquine, cycloquine, or pyronaridine was incubated with the activating system and glutathione. Calculation of the heat of formation of the drugs, however, demonstrated that amodiaquine, tebuquine, cycloquine, and pyronaridine readily undergo oxidation to their quinoneimine. None of the antimalarial compounds depleted the level of intracellular glutathione (1-300 microM) when incubated with neutrophils alone. Additionally, with the exception of tebuquine, no cytotoxicity below 100 microM was observed. In the presence of the full activating system, however, all compounds except chloroquine resulted in depletion of the level of glutathione and were cytotoxic. Pretreating the cells with glutathione and other antioxidants inhibited metabolism-dependent cytotoxicity. In summary, our data show that amodiaquine and related antimalarials containing a p-aminophenol moiety undergo bioactivation in vitro to chemically reactive and cytotoxic intermediates. In particular, pyronaridine, which is currently being investigated in humans, was metabolized to a compound which was toxic to neutrophils. Thus, the possibility that it will cause agranulocytosis in clinical practice cannot be excluded, and will require careful monitoring. PMID- 9860507 TI - NO release from NO donors and nitrovasodilators: comparisons between oxyhemoglobin and potentiometric assays volume 11, number 12, december 1998, pp 1393-1397 PMID- 9860506 TI - Metabolic activation of racemic and enantiomeric trans-8, 9-dihydroxy-8,9 dihydrodibenzo[a,l]pyrene (dibenzo[def,p]chrysene) to dibenzo[a,l]pyrene-bis dihydrodiols by induced rat liver microsomes and a recombinant human P450 1A1 system: the role of the K-region-derived metabolic intermediates in the formation of dibenzo[a,l]pyrene-DNA adducts. AB - Metabolic activation studies of dibenzo[a,l]pyrene (DB[a,l]P) (dibenzo[def,p]chrysene), an extremely potent environmental carcinogen, have been focused on metabolism at the fjord region, a region associated with high mutagenic and carcinogenic activities of the corresponding fjord-region DB[a,l]P 11,12-diol-13,14-epoxides. DB[a,l]P is metabolized by beta-naphthoflavone (BNF)- and 3-methylcholanthrene-induced rat liver microsomes and a recombinant human P450 1A1 system to two major dihydrodiols, the K-region dihydrodiol, DB[a,l]P-8,9 dihydrodiol (DB[a,l]P-8,9-diol), and the fjord-region dihydrodiol, DB[a,l]P-11,12 dihydrodiol. We have investigated the further metabolic activation of DB[a,l]P 8,9-diol by BNF-induced rat liver microsomes and a recombinant human P450 1A1 system with epoxide hydrolase to DB[a,l]P-bis-diols and to DNA adducts. (+/-) trans-DB[a,l]P-8,9-diol was synthesized and resolved into its enantiomers. Racemic trans-DB[a,l]P-8,9-diol was metabolized by BNF-induced rat liver microsomes to six metabolites: two diastereomers of trans,trans-DB[a,l]P 8,9:11,12-bis-diol, two diastereomers of trans,cis-DB[a,l]P-8,9:11,12-bis-diol, and two diastereomers of trans-DB[a,l]P-8,9:13,14-bis-diol as characterized by NMR, MS, and UV spectroscopy. Metabolic studies using both enantiomeric (-)- and (+)-trans-DB[a,l]P-8,9-diol further demonstrated that each diastereomer of trans,trans-DB[a,l]P-8,9:11, 12-bis-diol and trans-DB[a,l]P-8,9:13,14-bis-diol was comprised of two enantiomers. Similarly, incubations of enantiomeric or racemic trans-DB[a,l]P-8,9-diol with a recombinant human P450 1A1 system and epoxide hydrolase also gave the same two enantiomeric mixtures of diastereomers of trans,trans-DB[a,l]P-8,9:11,12-bis-diol and the same two enantiomeric mixtures of diastereomers of trans-DB[a,l]P-8, 9:13,14-bis-diol. This suggested that the microsomal oxidations of (-)- and (+)-trans-DB[a,l]P-8,9-diol were stereospecific. The stereospecific formation of enantiomers of trans-DB[a,l]P-8,9 diol from DB[a,l]P was examined using both BNF-induced rat liver microsomes and a recombinant human P450 1A1 system with epoxide hydrolase. Stereospecificity was observed as both metabolic systems favored the formation of (-)-trans-DB[a,l]P 8,9-diol by 8-9-fold. DNA adduct studies were undertaken using TLC/HPLC 32P postlabeling techniques. In the presence of a recombinant human P450 1A1 system with epoxide hydrolase, DB[a,l]P gave two groups of calf thymus DNA adducts. The group of later-eluting adducts were identified as arising from syn- and anti DB[a,l]P-11,12-diol-13,14-epoxides, while the more polar early-eluting adducts were derived, in part, from the further activation of trans-DB[a,l]P-8,9-diol. Our data indicate that, in P450 1A1-mediated microsomal incubations, DB[a,l]P is metabolized to trans-DB[a,l]P-8,9-diol which is further metabolized to DB[a,l]P bis-diols. trans-DB[a,l]P-8,9-diol is metabolically activated to intermediates that can bind to DNA and give DNA adducts similar to those observed with DB[a,l]P. These results indicate that DB[a,l]P can be metabolically activated by both fjord-region and K-region pathways. PMID- 9860775 TI - To E or not to E--how do we tell? PMID- 9860776 TI - Inhibiting tissue angiotensin-converting enzyme: a pound of flesh without the blood? PMID- 9860777 TI - Localization of a gene responsible for arrhythmogenic right ventricular dysplasia to chromosome 3p23. AB - BACKGROUND: Arrhythmogenic right ventricular dysplasia (ARVD), a familial cardiomyopathy occurring with a prevalence of 1 in 5000, is characterized by replacement of myocytes with fatty and fibrous tissue. Clinical manifestations include structural and functional abnormalities of the right ventricle and arrhythmias, leading to a sudden death rate of 2.5% per year. Four loci have been mapped, but no gene has been identified as yet. METHODS AND RESULTS: We identified a large family of >200 members with ARVD segregating as an autosomal dominant trait affecting 10 living individuals. The diagnosis of ARVD was based on international diagnostic criteria including history, physical examination, ECG, echocardiogram, right ventricular angiogram, endomyocardial biopsy, and 24 hour ambulatory ECG. Blood was collected for DNA from 149 family members. Analysis of 257 polymorphic microsatellite markers by genetic linkage excluded previously known loci for ARVD and identified a novel locus at 3p23. Analysis of an additional 20 markers further defined the region. A peak logarithm of the odds score of 6.91 was obtained with marker D3S3613 at theta=0% recombination. Haplotype analysis identified a shared region between markers D3S3610 and D3S3659 of 9. 3 cM. CONCLUSIONS: A novel locus for ARVD has been mapped to 3p23 and the region narrowed to 9.3 cM. Identification of the gene will allow genetic screening and a specific diagnosis for a disease with protean nonspecific findings. It should also provide insight fundamental to understanding cardiac chamber-specific gene expression and/or the mechanism of myocyte apoptosis observed in this disease. PMID- 9860778 TI - Prospective study of herpes simplex virus, cytomegalovirus, and the risk of future myocardial infarction and stroke. AB - BACKGROUND: It has been hypothesized that infection with either herpes simplex virus (HSV) or cytomegalovirus (CMV) is associated with atherogenesis. However, prospective data relating evidence of prior exposure to these agents with risks of future myocardial infarction (MI) and stroke are sparse. METHODS AND RESULTS: In a prospective, nested case-control study of apparently healthy men, the baseline prevalence of antibodies directed against HSV or CMV was similar among 643 men who subsequently developed a first MI or thromboembolic stroke and among 643 age- and smoking-matched men who remained free of reported vascular disease over a 12-year follow-up period. Specifically, the relative risks for future MI and stroke were 0.94 (95% CI, 0.7 to 1.2) for HSV seropositivity and 0.72 (95% CI, 0.6 to 0.9) for CMV seropositivity, after adjustment for other cardiovascular risk factors. These findings were not materially altered in comparisons of early versus late events or in analyses stratified by smoking status. There was no evidence of association between HSV or CMV antibodies and plasma concentration of C-reactive protein, a marker of inflammation that predicts vascular risk in this cohort. CONCLUSIONS: Among apparently healthy middle-aged men, IgG antibodies directed against HSV or CMV do not appear to be a marker for increased atherothrombotic risk. The observed possible inverse relationship of CMV with MI and stroke was unexpected and may well be due to chance, because the direction of association is not compatible with the a priori hypothesis based on proposed biological mechanisms or previous cross-sectional and retrospective data. PMID- 9860779 TI - Gene therapy for myocardial angiogenesis: initial clinical results with direct myocardial injection of phVEGF165 as sole therapy for myocardial ischemia. AB - BACKGROUND: We initiated a phase 1 clinical study to determine the safety and bioactivity of direct myocardial gene transfer of vascular endothelial growth factor (VEGF) as sole therapy for patients with symptomatic myocardial ischemia. METHODS AND RESULTS: VEGF gene transfer (GTx) was performed in 5 patients (all male, ages 53 to 71) who had failed conventional therapy; these men had angina (determined by angiographically documented coronary artery disease). Naked plasmid DNA encoding VEGF (phVEGF165) was injected directly into the ischemic myocardium via a mini left anterior thoracotomy. Injections caused no changes in heart rate (pre-GTx=75+/-15/min versus post-GTx=80+/-16/min, P=NS), systolic BP (114+/-7 versus 118+/-7 mm Hg, P=NS), or diastolic BP (57+/-2 versus 59+/-2 mm Hg, P=NS). Ventricular arrhythmias were limited to single unifocal premature beats at the moment of injection. Serial ECGs showed no evidence of new myocardial infarction in any patient. Intraoperative blood loss was 0 to 50 cm3, and total chest tube drainage was 110 to 395 cm3. Postoperative cardiac output fell transiently but increased within 24 hours (preanesthesia=4.8+/-0.4 versus postanesthesia=4.1+/-0.3 versus 24 hours postoperative=6. 3+/-0.8, P=0.02). Time to extubation after closure was 18.4+/-1.4 minutes; average postoperative hospital stay was 3.8 days. All patients had significant reduction in angina (nitroglycerin [NTG] use=53.9+/-10.0/wk pre-GTx versus 9.8+/-6.9/wk post-GTx, P<0.03). Postoperative left ventricular ejection fraction (LVEF) was either unchanged (n=3) or improved (n=2, mean increase in LVEF=5%). Objective evidence of reduced ischemia was documented using dobutamine single photon emission computed tomography (SPECT)-sestamibi imaging in all patients. Coronary angiography showed improved Rentrop score in 5 of 5 patients. CONCLUSIONS: This initial experience with naked gene transfer as sole therapy for myocardial ischemia suggests that direct myocardial injection of naked plasmid DNA, via a minimally invasive chest wall incision, is safe and may lead to reduced symptoms and improved myocardial perfusion in selected patients with chronic myocardial ischemia. PMID- 9860780 TI - TNK-tissue plasminogen activator compared with front-loaded alteplase in acute myocardial infarction: results of the TIMI 10B trial. Thrombolysis in Myocardial Infarction (TIMI) 10B Investigators. AB - BACKGROUND: Bolus thrombolytic therapy is a simplified means of administering thrombolysis that facilitates rapid time to treatment. TNK-tissue plasminogen activator (TNK-tPA) is a highly fibrin-specific single-bolus thrombolytic agent. METHODS AND RESULTS: In TIMI 10B, 886 patients with acute ST-elevation myocardial infarction presenting within 12 hours were randomized to receive either a single bolus of 30 or 50 mg TNK-tPA or front-loaded tPA and underwent immediate coronary angiography. The 50-mg dose was discontinued early because of increased intracranial hemorrhage and was replaced by a 40-mg dose, and heparin doses were decreased. TNK-tPA 40 mg and tPA produced similar rates of TIMI grade 3 flow at 90 minutes (62.8% versus 62.7%, respectively, P=NS); the rate for the 30-mg dose was significantly lower (54.3%, P=0.035) and was 65. 8% for the 50-mg dose (P=NS). A prespecified analysis of weight-based TNK-tPA dosing using median TIMI frame count demonstrated a dose response (P=0.001). Similar dose responses were observed for serious bleeding and intracranial hemorrhage, but significantly lower rates were observed for both TNK-tPA and tPA after the heparin doses were lowered and titration of the heparin was started at 6 hours. CONCLUSIONS: TNK tPA, given as a single 40-mg bolus, achieved rates of TIMI grade 3 flow similar to those of the 90-minute bolus and infusion of tPA. Weight-adjusting TNK-tPA appears to be important in achieving optimal reperfusion; reduced heparin dosing appears to improve safety for both agents. Together with the safety results from the parallel Assessment of the Safety of a New Thrombolytic: TNK-tPA (ASSENT I) trial, an appropriate dose of this single-bolus thrombolytic agent has been identified for phase III testing. PMID- 9860781 TI - Correlation of factor VIIa values with factor VII gene polymorphism, fasting and postprandial triglyceride levels, and subclinical carotid atherosclerosis. AB - BACKGROUND: Factor VII plays a pivotal role in coagulation. Factor VIIc levels were reported to be a risk factor for fatal coronary heart disease (CHD). Factor VIIc and VIIag levels were noted to be positively associated with plasma triglyceride (TG) levels and influenced by a VII gene polymorphism. The purpose of this study is to determine whether these associations are related to activated factor VII (factor VIIa). METHODS AND RESULTS: Fasting and 3.5-hour postprandial samples from 216 cases with subclinical atherosclerosis and 341 matched controls selected from the ARIC cohort were assayed for levels of factors VIIa, VIIc, and VIIag and TG, and factor VII codon 353 gene polymorphism. The level of factor VIIa was higher in Arg/Arg than in Arg/Gln+Gln/Gln genotypes, and the difference was in accord with that of factors VIIag and VIIc. However, the factor VIIa difference was statistically insignificant. Factor VIIa values were not correlated with fasting or 3.5-hour postprandial TG levels, nor were they associated with subclinical atherosclerosis. CONCLUSIONS: Factor VIIa levels, like factor VIIag and VIIc levels, are influenced by factor VII gene codon 353 polymorphism. However, unlike factor VIIag or VIIc, factor VIIa is not influenced by TG levels; none of these is associated with subclinical atherosclerosis. PMID- 9860782 TI - Increased formation of distinct F2 isoprostanes in hypercholesterolemia. AB - BACKGROUND: F2 isoprostanes are stable, free radical-catalyzed products of arachidonic acid that reflect lipid peroxidation in vivo. METHODS AND RESULTS: Specific assays were developed by use of mass spectrometry for the F2 isoprostanes iPF2alpha-III and iPF2alpha-VI and arachidonic acid (AA). Urinary excretion of the 2 F2 isoprostanes was significantly increased in hypercholesterolemic patients, whereas substrate AA in urine did not differ between the groups. iPF2alpha-III (pmol/mmol creatinine) was elevated (P<0.0005) in homozygous familial hypercholesterolemic (HFH) patients (85+/-5. 5; n=38) compared with age- and sex-matched normocholesterolemic control subjects (58+/ 4.2; n=38), as were levels of iPF2alpha-VI (281+/-22 versus 175+/-13; P<0.0005). Serum cholesterol correlated with urinary iPF2alpha-III (r=0.41; P<0.02) and iPF2alpha-VI (r=0. 39; P<0.03) in HFH patients. Urinary excretion of iPF2alpha III (81+/-10 versus 59+/-4; P<0.05) and iPF2alpha-VI (195+/-18 versus 149+/-20; P<0.05) was also increased in moderately hypercholesterolemic subjects (n=24) compared with their controls. Urinary excretion of iPF2alpha-III and iPF2alpha-VI was correlated (r=0.57; P<0.0001; n=106). LDL iPF2alpha-III levels (ng/mg arachidonate) were elevated (P<0.01) in HFH patients (0.32+/-0.08) compared with controls (0.09+/-0.02). The concentrations of iPF2-III in LDL and urine were significantly correlated (r=0.42; P<0.05) in HFH patients. CONCLUSIONS: Asymptomatic patients with moderate and severe hypercholesterolemia have evidence of oxidant stress in vivo. PMID- 9860783 TI - Clinical outcomes of therapeutic agents that block the platelet glycoprotein IIb/IIIa integrin in ischemic heart disease. AB - BACKGROUND: Several platelet glycoprotein (GP) IIb/IIIa receptor antagonists have been evaluated in clinical trials. We conducted a systematic overview (meta analysis) to assess the effect of these compounds on death, myocardial infarction (MI), and revascularization. METHODS AND RESULTS: ORs were calculated for 16 randomized, controlled trials of GP IIb/IIIa inhibitors. An empirical Bayesian random-effects model combined the outcomes of 32 135 patients. There was a significant mortality reduction by GP IIb/IIIa inhibitors at 48 to 96 hours, with an OR of 0.70 (95% CI, 0. 51 to 0.96; P<0.03), equivalent to a reduction of 1 death per 1000 patients treated. Mortality benefits at 30 days (OR, 0.87; 95% CI, 0. 74 to 1.02; P=0.08) and 6 months (OR, 0.97; 95% CI, 0.86 to 1.10; P=0.67) were not statistically significant. For the combined end point of death or MI, there was a highly significant (P<0.001) benefit for GP IIb/IIIa inhibitors at each time point. The 30-day OR was 0.76 (95% CI, 0.66 to 0.87), or 20 fewer events per 1000 patients treated. For the composite end point of death, MI, or revascularization, there was also a highly significant (P<0.001) benefit for GP IIb/IIIa inhibitors. At 30 days, the OR was 0.77 (95% CI, 0.68 to 0.86), or 30 fewer events per 1000 patients treated. The risk differences for death, death or MI, and composite outcomes were similar at 6 months, indicating a sustained absolute improvement. Similar benefit was seen when trials were subgrouped by therapeutic indication (percutaneous intervention versus acute coronary syndromes). CONCLUSIONS: Application of this new therapeutic class to clinical practice promises substantial benefit for both indications. PMID- 9860784 TI - Prognostic value of treadmill exercise testing: a population-based study in Olmsted County, Minnesota. AB - BACKGROUND: The prognostic value of treadmill exercise testing (TMET) has been studied in selected populations. The generalizability of these data to different populations and to women is uncertain. METHODS AND RESULTS: A retrospective, population-based cohort study of all persons (1452 men and 741 women) who underwent TMET in years 1987 to 1989 in Olmsted County, Minnesota, was undertaken. Individuals were followed up for all-cause mortality and cardiac events (cardiac deaths, nonfatal myocardial infarction, or congestive heart failure). Sex-specific analyses were performed to determine whether the predictors of outcome and the magnitude of the associations were similar in both sexes. In men, 77 deaths and 106 cardiac events occurred during 8956 person-years of observation; in women, 46 deaths and 54 cardiac events occurred during 4801 person-years of follow-up. Exercise-induced angina, ECG changes, and workload achieved on the TMET were strongly associated with all-cause mortality and cardiac events in both sexes, and the strength of the association was similar. After adjustment, workload was the only TMET variable associated with outcome. A higher workload was associated with a reduction in the risk of cardiac events and of all-cause mortality; the protective effect of exercise capacity was strong and was similar in both sexes. CONCLUSIONS: In this population-based cohort, exercise capacity was the TMET variable that exhibited the strongest association with all cause mortality and cardiac events. This protective effect of exercise capacity was observed in both sexes. PMID- 9860785 TI - Differential effects of quinaprilat and enalaprilat on endothelial function of conduit arteries in patients with chronic heart failure. AB - BACKGROUND: Chronic heart failure (CHF) is associated with endothelial dysfunction, including impaired flow-dependent (endothelium-mediated) dilation (FDD). We have previously shown that ACE inhibition improves endothelium-mediated vasodilation in healthy volunteers. The present study was designed to determine whether ACE inhibition improves the impaired FDD in patients with CHF. Because their affinity to tissue ACE may influence the ability of ACE inhibitors to affect endothelial function, we compared the effects of quinaprilat (high affinity to tissue ACE) and enalaprilat (low affinity to tissue ACE) on FDD in patients with CHF. METHODS AND RESULTS: High-resolution ultrasound and Doppler were used to measure radial artery diameter and blood flow in patients with CHF. The effects of intra-arterial infusion of quinaprilat 1.6 microg/min (n=15) and enalaprilat 5 microg/min (n=15) were determined at rest and during reactive hyperemia (causing endothelium-mediated dilation) before and after N-monomethyl-L arginine (L-NMMA) to inhibit endothelial synthesis of nitric oxide. Quinaprilat improved FDD by >40% (10.2+/-0.6% versus 6.9+/-0.6%; P<0.01), whereas enalaprilat had no effect. In particular, the part of FDD mediated by nitric oxide (ie, inhibited by L-NMMA) was increased by >100% with quinaprilat (5.6+/-0.5% versus 2.5+/-0.5%; P<0.01). Enalaprilat had no effect on FDD even when it was infused twice in the same dose (5 microg/min) and up to 30 microg/min. The effect of sodium nitroprusside on radial artery diameter and blood flow was similar in patients treated with quinaprilat, enalaprilat, and placebo. CONCLUSIONS: Quinaprilat improves FDD in patients with CHF as the result of increased availability of nitric oxide, whereas enalaprilat does not. This observation suggests that intrinsic differences exist between quinaprilat and enalaprilat that determine the ability to improve endothelium-mediated vasodilation, ie, their different affinity to tissue ACE. PMID- 9860787 TI - Dipyridamole stress echocardiography for risk stratification in hypertensive patients with chest pain. AB - BACKGROUND: The noninvasive prognostic assessment of coronary artery disease (CAD) in hypertensive patients represents an unresolved task to date. In this study, we investigated the value of dipyridamole stress echocardiography in risk stratification of hypertensive patients with chest pain and unknown CAD. METHODS AND RESULTS: Dipyridamole stress echocardiography was performed in 257 hypertensives (110 men; age, 63+/-9 years) complaining of chest pain and without a history of CAD. No major complications occurred. Four tests were interrupted prematurely because of side effects, with 98. 4% feasibility of test. A positive echocardiographic response was found in 72 patients (27 during the low-dose [0.56 mg/kg]). During the follow-up (32+/-18 months), 27 cardiac events occurred: 3 deaths, 8 infarctions, and 16 cases of unstable angina. Moreover, 27 patients underwent coronary revascularization. At multivariate analysis, the positive echocardiographic result (OR, 5.5; 95% CI, 1.4 to 16.6) was the only predictor of hard cardiac events (death, infarction). Considering spontaneous cardiac events (death, infarction, and unstable angina) as end points, the positive echocardiographic result (OR, 4.2; 95% CI, 1.8 to 9.6) and family history of CAD (OR, 4.2; 95% CI, 1.5 to 6. 9) were independently associated with prognosis. The 3-year survival rates for the negative and the positive populations were, respectively, 97% and 87% (P=0.0019) considering hard cardiac events and 96% and 74% (P=0.0000) considering spontaneous cardiac events. CONCLUSIONS: Dipyridamole stress echocardiography is safe, highly feasible, and effective in risk stratification of hypertensives with chest pain and unknown CAD. At present, it represents an attractive option for prognostic assessment of this clinically defined population. PMID- 9860786 TI - Early impairment of renal hemodynamic reserve in patients with asymptomatic heart failure is restored by angiotensin II antagonism. AB - BACKGROUND: The early/asymptomatic stages of heart failure (HF) are characterized by sodium retention secondary to derangement of sodium reabsorption at the proximal nephron level. Because this phenomenon is reversed by ACE inhibition, abnormalities of renal sodium handling may depend on intrarenal changes of angiotensin II (AII)/nitric oxide (NO) levels. Renal hemodynamic reserve (ie, the glomerular vasodilatory response to amino acid infusion) has been proposed as a reliable test to assess in vivo AII/NO balance. METHODS AND RESULTS: In this study, the effects of 6 weeks of treatment with 5 mg/d of enalapril or with 50 mg/d of losartan on systemic hemodynamics and renal function were assessed, at baseline and after amino acid infusion (AA), in patients with mild HF (NYHA class I) and in healthy volunteers. Untreated HF patients showed a basal renal function comparable to that of healthy subjects. After AA, glomerular filtration rate and renal plasma flow significantly increased in healthy subjects (+29.0% and +30.4%, respectively), whereas no vasodilatory response was observed in HF. Although they did not affect basal renal hemodynamics, both enalapril and losartan restored a normal response to AA in HF patients. Blood pressure and heart rate were comparable in HF subjects and healthy subjects at baseline and were not modified by either treatment. Left ventricular ejection fraction was depressed in HF but did not change after either drug. Urinary excretions of cGMP and nitrate (indexes of NO activity in the kidney), comparable in healthy subjects and in HF patients, were unchanged by either enalapril or losartan and did not correlate with renal reserve. CONCLUSIONS: (1) Renal functional reserve is absent in patients with early/asymptomatic HF and normal renal function and (2) both enalapril and losartan restore a normal vasodilatory response to AA in these patients without affecting basal systemic and renal hemodynamics. These data suggest a major role of AII in the development of early abnormalities in patients with HF. PMID- 9860788 TI - Electrophysiological characteristics of the human atria after cardioversion of persistent atrial fibrillation. AB - BACKGROUND: In animal models, induced atrial fibrillation shortens the atrial effective refractory period (ERP) and reverses its physiological adaptation to rate. It is not clear whether this process, known as "electrical remodeling," occurs in humans. METHODS AND RESULTS: We determined the ERPs, at 5 pacing cycle lengths (300 to 700 ms) and in 5 right atrial sites, after internal cardioversion of chronic atrial fibrillation in 25 patients (14 in pharmacological washout and 11 on amiodarone). The ERPs were 195.5+/-18.8 ms in the washout and 206.3+/-17.9 ms in the amiodarone patients (P<0.0001). ERPs were closely correlated with the stimulation rates (r=0.95 in the washout and r=0.94 in the amiodarone group), and slope values indicating a normal (>/=0.07) or nearly normal (0.05 to 0.06) adaptation of ERP to rate were found in 77% of the 84 paced sites. The mean ERP was shorter in the lateral wall (198.1+/-17.9 ms) than in the atrial roof (203.3+/-21.5 ms) and in the septum (210.5+/-20.0 ms) (P<0.03). After 4 weeks of sinus rhythm, the mean ERP, determined again in 8 patients (4 in wash-out and 4 on amiodarone), was significantly increased compared with the basal study (221. 4+/-21.4 versus 197.8+/-18.3 ms, P<0.0001). CONCLUSIONS: After cardioversion of chronic atrial fibrillation, (1) atrial ERP adaptation to rate was normal or nearly normal in the majority of the cases, (2) a significant dispersion of refractoriness between different right atrial sites was present, and (3) ERPs were significantly increased after 4 weeks of sinus rhythm in both washout and amiodarone patients. PMID- 9860789 TI - Prospective study of atherosclerotic disease progression in the renal artery. AB - BACKGROUND: The aim of this study was to determine the incidence of and the risk factors associated with progression of renal artery disease in individuals with atherosclerotic renal artery stenosis (ARAS). METHODS AND RESULTS: Subjects with >/=1 ARAS were monitored with serial renal artery duplex scans. A total of 295 kidneys in 170 patients were monitored for a mean of 33 months. Overall, the cumulative incidence of ARAS progression was 35% at 3 years and 51% at 5 years. The 3-year cumulative incidence of renal artery disease progression stratified by baseline disease classification was 18%, 28%, and 49% for renal arteries initially classified as normal, <60% stenosis, and >/=60% stenosis, respectively (P=0.03, log-rank test). There were only 9 renal artery occlusions during the study, all of which occurred in renal arteries having >/=60% stenosis at the examination before the detection of occlusion. A stepwise Cox proportional hazards model included 4 baseline factors that were significantly associated with the risk of renal artery disease progression during follow-up: systolic blood pressure >/=160 mm Hg (relative risk [RR]=2.1; 95% CI, 1.2 to 3.5), diabetes mellitus (RR=2.0; 95% CI, 1.2 to 3.3), and high-grade (>60% stenosis or occlusion) disease in either the ipsilateral (RR=1.9; 95% CI, 1.2 to 3.0) or contralateral (RR=1.7; 95% CI, 1.0 to 2.8) renal artery. CONCLUSIONS: Although renal artery disease progression is a frequent occurrence, progression to total renal artery occlusion is not. The risk of renal artery disease progression is highest among individuals with preexisting high-grade stenosis in either renal artery, elevated systolic blood pressure, and diabetes mellitus. PMID- 9860790 TI - In vivo evaluation of Fontan pathway flow dynamics by multidimensional phase velocity magnetic resonance imaging. AB - BACKGROUND: Hemodynamic efficiency of Fontan circulation is believed to be a major determinant of outcome. Prior research on flow dynamics in different modifications of Fontan circulation used in vitro models and computer-based simulation. This study was designed to compare in vivo flow dynamics in the systemic venous pathway between patients with atriopulmonary anastomosis (APA) and those with total cavopulmonary connection (TCPC). METHODS AND RESULTS: Multidimensional phase-velocity magnetic resonance imaging (PV-MRI) studies were performed on 10 patients who had undergone a modified Fontan operation (5 with TCPC and 5 with APA) and were free of symptoms. The groups were comparable in terms of age and body surface area. The interval since surgery was longer for APA than for TCPC subjects. In each subject, the phase-velocity data sets were used to generate dynamic velocity-vector maps and to calculate quantitative flow indices describing the 3-dimensional blood-flow patterns throughout the cardiac cycle at the widest diameter of the Fontan pathway. Mean flow rate was comparable between groups. Velocity-vector maps showed areas of flow reversal, flow stagnation, and circular flow within APA but not TCPC pathways. Analysis of quantitative flow indices showed that compared with the APA group, flow velocities in the TCPC patients were significantly higher (mean velocity, 14+/-6 cm/s versus 5+/-3 cm/s; P=0.02), less variable (coefficient of variation, 19+/-2% versus 37+/-3.5%; P<0.0001), and more unidirectional (degree of unidirectionality, 89+/-7% versus 71+/-12%; P=0.03). APA pathways were significantly more dilated than were TCPC pathways (P<0.01) and showed a trend toward larger diameter with increased interval since surgery (R2=0.6, P=0.09). Fontan pathway dilatation correlated with flow velocity variability (R2=0.57, P=0.01) and inversely with flow unidirectionality (R2=0.75, P=0.001). CONCLUSIONS: Blood flow patterns are more organized and uniform in TCPC than in APA pathways and are significantly influenced by pathway diameter. We speculate that TCPC may result in a more hemodynamically efficient circulation than APA because of differences in pathway dimension and uniformity. PMID- 9860791 TI - Restoration of E2F expression rescues vascular endothelial cells from tumor necrosis factor-alpha-induced apoptosis. AB - BACKGROUND: Normally, quiescent endothelial cells (EC) line the inner surface of arteries and protect against thrombosis and neointimal growth. A variety of noxious stimuli, including balloon angioplasty, may compromise EC integrity, thereby initiating proliferation and triggering the local release of cytokines, including tumor necrosis factor-alpha (TNF-alpha). METHODS AND RESULTS: In vivo blockade of TNF-alpha using a soluble receptor molecule results in accelerated reendothelialization at sites of balloon angioplasty, suggesting an important physiological role of TNF-alpha in attenuating regrowth of endothelium after balloon angioplasty. Our studies reveal that TNF-alpha, an apoptosis-inducing cytokine, induces G1 cell-cycle arrest in proliferating EC. Quiescent EC are relatively immune to TNF-induced apoptosis versus proliferating EC, which display repression of the E2F transcription factor coincident with TNF-induced apoptosis and cell-cycle arrest. We also show that in this setting, E2F overexpression exerts a survival effect in proliferating EC and restores cell-cycle progression, in direct contrast to results of prior reports, which revealed that deregulated expression of E2F in normally cycling cells induces apoptosis. CONCLUSIONS: These data demonstrate that TNF-induced apoptosis is highly dependent on cell-cycle activity and that E2F can function as survival factor under certain conditions. PMID- 9860792 TI - Platelet-derived 12-hydroxyeicosatetraenoic acid plays an important role in mediating canine coronary thrombosis by regulating platelet glycoprotein IIb/IIIa activation. AB - BACKGROUND: In the thrombotic process of acute coronary syndromes, the pathophysiological role of thromboxane A2 via cyclooxygenase is well established; however, the role of 12-HETE via 12-lipoxygenase is little known. Therefore, we used OPC-29030, a novel specific inhibitor of 12-HETE synthesis, to test whether platelet-derived 12-HETE is involved in mediating cyclic flow variations (CFVs) and platelet aggregation in stenosed and endothelium-injured canine coronary arteries. METHODS AND RESULTS: After developing CFVs, dogs received a vehicle or OPC-29030 intravenously. Plasma and intraplatelet 12-HETE levels increased after CFVs. OPC-29030 but not vehicle reduced CFVs, which was associated with decreases in plasma and intraplatelet 12-HETE levels. Cessation of OPC-29030 restored CFVs in association with increases in plasma and intraplatelet 12-HETE levels. ADP and U46619 induced ex vivo platelet 12-HETE production and aggregation. After OPC 29030 administration, the ADP- and U46619-induced increases in ex vivo platelet 12-HETE production and aggregation were inhibited significantly. Platelet aggregation was linearly correlated with platelet 12-HETE production. OPC-29030 suppressed activation of human platelet glycoprotein IIb/IIIa. CONCLUSIONS: OPC 29030 reduced intraplatelet 12-HETE levels, resulting in the inhibition of coronary thrombosis in vivo in dogs. OPC-29030 inhibited human platelet glycoprotein IIb/IIIa activation in vitro. Thus, platelet-derived 12-HETE may play an important role in mediating thrombotic process. PMID- 9860793 TI - Offspring of normal and diabetic rats fed saturated fat in pregnancy demonstrate vascular dysfunction. AB - BACKGROUND: Disturbances of the in utero environment may "program" for disease in later life. In this study, we determined whether dietary fat supplementation and/or diabetes in pregnancy can adversely affect vascular function in the offspring. METHODS AND RESULTS: Female Sprague-Dawley rats were fed a breeding diet or a diet high in saturated fat (30% wt/wt) for 10 days before mating, throughout pregnancy, and postpartum. Endothelium-dependent relaxation to acetylcholine was blunted in isolated femoral arteries of 15-day-old weanling pups from dams fed the 30%-fat diet. Endothelial dysfunction and enhanced constrictor responses to norepinephrine were also observed in an additional study of 60-day-old offspring of dams fed 20% saturated fat. Rats with streptozotocin induced diabetes were also fed saturated fat during pregnancy. Femoral arteries from their 15-day-old offspring showed impairment of endothelium-dependent dilation and enhanced constrictor responses to norepinephrine and the thromboxane mimetic U46619 compared with young offspring of high-fat-fed normal dams. The 30% fat diet was also deleterious to vascular function in the maternal diabetic animals when assessed in mesenteric arteries 16 days postpartum. CONCLUSIONS: A high-fat diet in pregnancy led to vascular dysfunction in rat weanlings and young adult offspring. Vascular function further deteriorated in weanlings if the maternal rat was diabetic. PMID- 9860794 TI - Coordinate interaction between ATP-sensitive K+ channel and Na+, K+-ATPase modulates ischemic preconditioning. AB - BACKGROUND: We reported that digoxin abolishes the infarct size (IS)-limiting effect of ischemic preconditioning (IPC). Because ATP-sensitive K+ (KATP) channels are involved in IPC, we studied whether Na+,K+-ATPase and KATP channels functionally interact, thereby modulating IPC. METHODS AND RESULTS: Rabbits received 30 minutes of coronary artery occlusion followed by 3 hours of reperfusion. IPC was elicited by 5 minutes of occlusion followed by 10 minutes of reperfusion. The IS, expressed as a percentage of the area at risk, was 40.2+/ 2.8% in control and 39.8+/-5.0% in digoxin pretreatment rabbits. Both IPC and pretreatment with cromakalim, a KATP channel opener, reduced IS to 11.8+/-1.8% and 13.4+/-2.6% (P<0. 05 versus control). Digoxin abolished the reduction in IS induced by IPC (33.5+/-3.3%), whereas it did not change that induced by cromakalim (18.8+/-3.0%). In patch-clamp experiments, digoxin was found to inhibit the opening of KATP channels in single ventricular myocytes in which ATP depletion had been induced by metabolic stress. In contrast, digoxin had little effect on the channel opening induced by cromakalim. Moreover, the inhibitory action of digoxin on channel activities was dependent on subsarcolemmal ATP concentration. CONCLUSIONS: The IS-limiting effect of IPC is modulated by an interaction between KATP channels and Na+,K+-ATPase through subsarcolemmal ATP. PMID- 9860795 TI - Preserved vasodilator effect of bradykinin in dogs with heart failure. AB - BACKGROUND: In heart failure (HF), vasoconstrictor systems are activated and endothelium-derived vasodilation is blunted. Bradykinin, a potent vasodilator, may play an important role in this setting. However, it is not known whether its vasodilator effect is modified in HF. METHODS AND RESULTS: Fourteen chronically instrumented dogs were studied in the control state and in pacing-induced HF (250 bpm for 3 weeks). The dose-dependent decrease in mean aortic pressure (MAP) induced by acetylcholine was significantly blunted in HF. In contrast, in both control and HF, bradykinin infusion caused similar dose-dependent decreases in MAP and increases in cardiac output (CO). This vasodilator effect of exogenous bradykinin was potentiated similarly in both states by enalaprilat, which blocks both angiotensin conversion and bradykinin degradation. For evaluating the role of endogenous bradykinin, the effects of enalaprilat were compared with those of ciprokiren, a pure renin inhibitor. In control, ciprokiren did not produce any effect. Enalaprilat, however, produced a significant decrease in MAP and a significant increase in CO, which were attributed to the inhibition of bradykinin degradation, because these effects were absent after pretreatment with Hoe 140 (a bradykinin B2 receptor antagonist). In contrast, in HF, vasodilator effects of ciprokiren were observed, but enalaprilat produced larger changes in MAP and CO, and after Hoe 140, the hemodynamic effects of enalaprilat were significantly decreased, showing the effects of endogenous bradykinin, which were similar to those measured in control. CONCLUSIONS: In this model of HF with a blunted endothelium-derived vasodilation, the vasodilator effects of exogenous and endogenous bradykinin are preserved. These results suggest that bradykinin may play an important role in HF, in which vasoconstriction is present and endothelium-dependent vasodilation is blunted. PMID- 9860796 TI - Mechanoenergetic alterations during the transition from cardiac hypertrophy to failure in Dahl salt-sensitive rats. AB - BACKGROUND: The time course and mechanisms of altered mechanoenergetics and depressed cross-bridge cycling in hypertrophied and failing myocardium are uncertain. METHODS AND RESULTS: We studied mechanoenergetics in Dahl salt sensitive (DS) rats fed high-salt diet (HS) for 6 (HS-6) and 12 (HS-12) weeks to produce compensated hypertrophy and failure. The slope of the end-systolic pressure-volume relation (E'max) was similar in HS-6 and low-salt controls (LS 6), but reduced in HS-12 compared with controls (LS-12). Efficiency [1/slope of oxygen consumption (&f1;O2)-pressure-volume area (PVA) relation] was similar in HS-6 and LS-6 but higher in HS-12 versus LS-12 (59+/-16% versus 44+/-7%, P<0.05). Economy [1/slope of the force-time integral (FTI)-&f1;O2 relation] was similar in HS-6 and LS-6 but higher in HS-12 versus LS-12 (218+/-123 versus 74+/-39x10(3) g. s. mL O2-1. g; P<0.05). Compared with controls, myofibrillar ATPase activity was reduced by 24% in HS-6 and 44% in HS-12. V3 Isomyosin was increased in HS-6 (40+/ 12% versus 9+/-8%; P<0.05) and further increased in HS-12 (76+/-10% versus 22+/ 18%; P<0.05). Hypothyroid LS-12 rats had 100% V3 isomyosin, yet efficiency, economy, and ATPase values were intermediate between LS-12 and HS-12. HS-12 rats demonstrated increased troponin T3 isoform (17+/-2 versus 23+/-2%, P<0.05). There were no changes in troponin I or tropomyosin isoforms. However, the proportion of phosphorylated troponin T was reduced in HS-12 versus LS-12 hearts (P<.001). CONCLUSIONS: In DS rats, the transition to failure is associated with depressed E'max and increased efficiency and economy. These findings are linked to myofibrillar ATPase activity and suggest that mechanisms other than isomyosin switching are important determinants of ventricular energetics. A troponin T isoform switch is one potential mechanism. PMID- 9860797 TI - Images in cardiovascular medicine. Combined angiography and three-dimensional computed tomography for assessing systemic-to-pulmonary collaterals in pulmonary atresia with ventricular septal defect. PMID- 9860798 TI - Methylenetetrahydrofolate reductase mutation and coronary artery disease. PMID- 9860799 TI - Safety of calcium channel blockers in cardiovascular disease. PMID- 9860800 TI - Pravastatin and coronary heart disease. PMID- 9860801 TI - Extended mortality benefit of early postinfarction reperfusion. PMID- 9860802 TI - Diagnosis and management of infective endocarditis and its complications. PMID- 9860803 TI - Molecular chaperones: biology and prospects for pharmacological intervention. PMID- 9860804 TI - Inflammatory mediators of asthma: an update. PMID- 9860805 TI - Glycine and N-methyl-D-aspartate receptors: physiological significance and possible therapeutic applications. PMID- 9860806 TI - Mechanisms of therapeutic activity for gallium. PMID- 9860807 TI - Myogenic and neurogenic factors in the control of pyeloureteral motility and ureteral peristalsis. PMID- 9860808 TI - G protein regulation of potassium ion channels. PMID- 9860809 TI - Zoophilic and geophilic fungi as a cause of skin disease in farmers. AB - The impact of microscopic fungi on the farmers' health seems to be underestimated. In the present article an overview of fungi as pathogens is presented with reference to occupational hygiene in agriculture and related areas. The infection may be transmitted from infected humans, animals, plants or soil. To date, little epidemiological data on fungal skin disease in farmers is available. Epidemiological studies from Poland suggest that mycoses are the most prevalent skin diseases in farmers, and may be present even in over 20% of the population. Working conditions on farms greatly enhance the development of fungal infections. Farmers spend most of their working time in humid conditions, wearing rubber boots for long hours, etc. Another professional groups at higher risk for developing a fungal disease are animal feeders, foresters, grave-diggers and veterinarians as well as employees working in the food industry. Besides infection, fungi may also cause non-invasive forms of skin disease, as dermato mycotoxicosis professionalis or alternariosis. Criteria for classifying a case of mycosis as occupational disease are also discussed. PMID- 9860810 TI - Aspects of dental health in adult rural population in Poland. AB - Using the literature on the subject, an assessment was made of the state of the oral health in rural population: dental state, periodontal diseases, state and needs for prosthetic rehabilitation. The articles presents possible causes of this state of things. PMID- 9860811 TI - Toxicity of dermally applied alpha-cypermethrin in rats. AB - The aim of the study was to assess the immunotoxic effect of dermally applied alpha-cypermethrin in rats based on phagocytic and bactericidal activity of neutrophils of peripheral blood, and the general toxic effect based on histological and ultrastructural examination of internal organs. The preparation was dermally applied in doses of 50 mg/kg and 250 mg/kg. It was administered to the tail skin of female Wistar rats, 4 hours daily for 28 days. After the experiment, the animals were anaesthetized and heart blood was taken in order to evaluate the activity of granulocyte system. The following organs were taken for histological examinations: brain, lung, heart, liver, spleen, kidneys, thymus and lymphatic nodes. Lung, liver, kidney and heart were used for ultrastructural studies. The results of the study showed that bactericidal and phagocytic activity of neutrophils was stimulated after administration of 50 mg/kg alpha cypermethrin. Dermal application of the preparation resulted in slight histological changes in liver, kidney, lung and brain. Pathological changes in heart were observed only on the level of ultrastructure. PMID- 9860812 TI - Major medical and social needs of disabled rural inhabitants. AB - A considerable increase in the number of the disabled has been observed in Poland during the last two decades, especially in rural areas, and constitutes a serious social and economic problem. It is therefore necessary to continue work on the detailed definition of disability and to develop new disability qualification classification methods (evaluation of the degree of invalidity). The all-Polish study of the state of health of rural population has considerably extended the knowledge of the problems experienced by this population group. The survey covered 1,491 people, including 779 females (52.2%) and 712 males (47.8%). The primary aim of the study was the qualitative and quantitative analysis of the medical and social situation of disabled adult rural inhabitants. The paper presents the basis on which respondents were classified as disabled, an analysis of their health and social situation, and analysis of their medical and social needs. The features were determined which distinguish subpopulations of those with legally ascribed categories of invalidity from those who have no legal decision concerning invalidity, and factors which distinguish farmers from non farmers. The most frequent causes of disability were cardiovascular diseases, followed by diseases of the musculoskeletal system and connective tissue, diseases of the nervous system and sense organs, and respiratory diseases. As much as 93.7% of the total number of the disabled required diagnostic and treatment procedures. A great demand for specialist treatment was observed among the youngest disabled aged 20-34. It was noted that the provision of orthopaedic and rehabilitation equipment, as well as of auxiliary aids, was highly insufficient from the aspect of both quality and quantity. Only 17% of the total number of disabled who expressed needs (130 out of 765 persons) were provided with such equipment. Only 8 people had a full range of technical adjustments in their dwellings. The disabled mentioned the following problems which significantly disturbed their functioning in everyday life: material difficulties, need for providing care for another disabled member of the family, the lack of independent lodging and proper employment. The study shows that the health, social and economic situation of the disabled rural inhabitants is very difficult. Further studies of this problem are needed, as well as the organization of medical, rehabilitation and social aid in this micro-environment. PMID- 9860813 TI - Exposure to dust among agricultural workers. AB - The authors present results of studies of occupational exposure to dust in agriculture, conducted for the first time from the aspect of work site. Two work sites, typical of Polish agriculture, were considered: a tractor driver on a large state owned farm and a farmer on a private family farm. The studies covered all occupational activities performed within the annual work cycle. The results of the studies showed that the working conditions in agriculture were hazardous and exposure to dust was of a changeable character. This is due to the varied levels of dustiness according to the type of occupation and unequal distribution of working time. PMID- 9860814 TI - Protein profile of the allergenic pollen of Ipomoea fistulosa L.--a comparative study. AB - The pollen of Ipomoea fistulosa L. is an important aeroallergen of India. The pollen of this plant was collected from full bloomed flowers growing in different places of West Bengal, India. Protein content and profile were studied by SDS PAGE. Skin-prick tests with pollen antigens of all the samples were also performed. Considerable variation in the protein content and profile was noted with the highest protein content in the collected sample of Calcutta showing highest number of protein bands (10) designated as IF1 to IF10 with their weights ranging between 31 kDa to 89 kDa and 3 bands (IF8, IF9 and IF10) having molecular weights less than 29 kDa. Skin-prick tests also revealed highest degree of sensitivity to the Calcutta sample giving positive response in 52% of the patients. Skin reactivity ranged between 1+ to 3+. PMID- 9860815 TI - Calves as a potential reservoir of Cryptosporidium parvum and Giardia sp. AB - Studies on cryptosporidiosis and giardiosis were carried out between March and April 1997 on 75 calves from 9 selected farms of Wielkopolska macroregion. Faecal specimens from calves, 3-13 days old, were screened for oocysts of C. parvum using Ziehl-Neelsen staining and both for oocysts of C. parvum and cysts of Giardia sp. using direct immunofluorescent (MerIFluor Cryptosporidium/Giardia) assay. The oocysts of C. parvum assessed by Ziehl-Neelsen staining were revealed on 6 (67%) of 9 farms examined. The prevalence of infection ranged from 20-88%, and in some farms intensity of oocyst shedding was very high. However, in 35 calves assessed for mixed infections of C. parvum and Giardia sp., oocysts of Cryptosporidium were found in 18 (51%) calves and cysts of Giardia sp. were detected in 5 (14%) of 35 calves. Only in one calf was found coinfection with both parasites. The intensity of Giardia sp. infection was extremely low. Histological examination of the gut sections from immunosuppressed BALB/c mice experimentally infected with C. parvum isolates from calves revealed endogenous stages of C. parvum on the brush border of the ileum. The high prevalence and intensive shedding of cryptosporidial oocysts by calves in farms examined in this study suggests that naturally infected calves may be significant reservoirs for C. parvum infections in man and wild animals. PMID- 9860816 TI - Working environment conditions in rural areas according to psychosocial indices. AB - The aim of this work was to study psychosocial working environment factors among farmers and other people living in rural areas. The study was carried out as a cross-section investigation. All persons visiting local occupational health service centres for a health check up have been asked to answer an inquiry which was based on the Karasek-Theorell questionnaire on job strain. Five extra items on worry about the future were added. The questionnaire was completed by over 3,800 persons. Three of four indices showed significant difference with respect to sex. Women experienced less stimulance at work, authority over work and had a greater fear of the future. Farmers had a significantly higher index for psychological demands, stimulance at work as well as authority over work than other occupational groups. The index for authority over work was very high in comparison with presented results for different occupations in other studies. With respect to worry about the future, the farmers had a significantly higher index than nearly all the other occupational groups. The low risk of coronary heart disease (CHD) among farmers reported in other studies can probably be related to good psychosocial working environment as measured by the indices in this study as well as other known life style factors. PMID- 9860817 TI - Effects of exposure to grain dust in Polish farmers: work-related symptoms and immunologic response to microbial antigens associated with dust. AB - Medical examinations were performed in a group of 76 Polish farmers heavily exposed to grain dust during harvesting and threshing, and in a group of 63 healthy urban dwellers not exposed to organic dusts (controls). The examinations included: interview concerning the occurrence of respiratory disorders and work related symptoms, physical examination, lung function tests, and allergological tests comprising skin prick test with 4 microbial antigens associated with grain dust and agar-gel precipitation test with 12 microbial antigens. As many as 34 farmers (44.7%) reported the occurrence of work-related symptoms during harvesting and threshing. The most common was dry cough reported by 20 individuals (26.3%). Dyspnoea was reported by 15 farmers (19.7%), tiredness by 12 (15.7%), chest tightness by 8 (10.5%), plugging of nose and hoarseness by 5 each (6. 5%). No control subjects reported these work-related symptoms. The mean spirometric values in the examined group of farmers were within the normal range, but a significant post-shift decrease of these values was observed after work with grain. The farmers showed a frequency of the positive early skin reactions to environmental allergens in the range of 10.8 - 45.5%, and a frequency of positive precipitin reactions in range of 3.9 - 40.8%. The control group responded to the majority of allergens with a significantly lower frequency of positive results compared to the farmers. The obtained results showed a high response of grain farmers to inhalant microbial allergens and indicate a potential risk of occupational respiratory diseases (such as allergic alveolitis, asthma, Organic Dust Toxic Syndrome) among this population PMID- 9860818 TI - Laboratory simulation of splashes and spills of organophosphate insecticides on chemically protective gloves used in agriculture. AB - Agricultural workers rely on chemically protective gloves for protection from dermal exposure to insecticides. In Australia the most widely used gloves are manufactured from polyvinyl chloride or nitrile butadiene rubber. During insecticide application splashes and spills frequently occur on the external surfaces of gloves which may compromise the integrity of the membrane. Interaction of two organophosphate insecticides, chlorpyrifos (Lorsban 500 EC(R)) and diazinon (Jetdip(R)), with glove surfaces was investigated in laboratory conditions. The external surface of gloves was treated with concentrated insecticides for one minute and diluted and concentrated insecticides for 24, 36 and 48 hours and later examined by environmental scanning electron microscopy. Two classes of defects, cavities and convexities, were evident in the polyvinyl chloride gloves following all treatments, whereas cracking was significant in the nitrile butadiene rubber gloves after 24 hours. In addition, X-ray energy dispersive microanalysis was used to evaluate chemical changes on the glove surfaces. Phosphorus and sulfur were useful indicators for organophosphate retention over specific time frames. Results corroborated the need for more robust chemically protective gloves to be developed for routine agricultural use. PMID- 9860819 TI - Study on exposure of pig farm workers to bioaerosols, immunologic reactivity and health effects. AB - Occupational inhalation of organic dust may be a cause of numerous symptoms and diseases. Organic dust consists of various biological compounds which induce inflammatory reactions in the lungs on an immunotoxic or allergic basis. Bacteria and their metabolites, moulds and their spores, mycotoxins, glucans, and other still unidentified substances, can be potential aetiologic factors of diseases caused by organic dust. The aim of this study was to determine the concentration and composition of airborne microflora in typical Polish swine buildings and to assess the health conditions of the workers employed within them. Total concentration of microorganisms in the air of five examined swine breeding farms ranged from 613.7-1246.7 x 10(3) cfu/m3 (mean value 930.6 x 10(3) cfu/ m3). The examination of 53 employees working in the swine buildings included their medical history, physical examination, spirometry, and allergological tests. Work-related symptoms were reported by 31 (58.5%) of the subjects. No abnormal findings were present upon physical and spirometric examinations. The results suggest the common occurrence of work-related respiratory disease in swine workers, mostly corresponding to the Organic Dust Toxic Syndrome (ODTS). PMID- 9860820 TI - Seroepidemiologic study on tick-borne encephalitis among forestry workers and farmers from the Lublin region (eastern Poland). AB - The paper presents the results of seroepidemiologic studies concerning tick-borne encephalitis (TBE) in 1,583 persons (1,261 forestry workers and 233 farmers) from the Lublin region (eastern Poland) occupationally exposed to ticks and in 130 healthy blood donors (a control group). The mean percentage of seropositive reactions in forestry workers amounted to 19.8% and in farmers 32.0%. Based on 5 year research (1994-1998) conducted in 5 districts of the Lublin region, the existence of endemic foci of TBE was detected in the district of Bia a Podlaska, on the areas of Radzyn Podlaski and Parczew, where the percentage of seropositive reactions in forestry workers exceeded 50%. Statistical analysis showed that the frequency of seropositive reactions in forestry workers and farmers was significantly greater compared to control group (p < 0.001 and p < 0.05, respectively). It indicates that these groups are occupationally exposed to TBE virus. In the years 1994-98, a total of nine clinical cases of TBE (acute neuroinfection) in forestry workers and fourteen clinical cases in farmers were confirmed serologically. The effectiveness of specific immunization against TBE was proved on the basis of 100% seroconversion in 56 earlier seronegative forestry workers. The obtained results proved that forestry workers and farmers in Poland are under increased risk of infection with TBE virus. PMID- 9860821 TI - Seroepidemiologic study on Lyme borreliosis in the Lublin region. AB - The frequency of anti-Borrelia burgdorferi antibodies in human sera of various groups of people from the Lublin region was studied. In the indirect immunofluorescence test (IFT) sera from 836 forestry workers and 56 farmers occupationally exposed to ticks were examined. Fifty healthy blood donors from the city of Lublin were examined as a control group. Forestry workers showed positive response in 26%, whereas farmers in 11% and the control group in 6%. In ELISA test, sera from 44 forestry workers, 217 farmers, 458 patients from the dermatologic and neurologic clinics, and 50 blood donors (controls) were examined. A positive response was found in 38.6% of forestry workers, 28.1% of farmers, 12.2% of patients of the neurologic clinic, 27.1% of patients of the dermatologic clinic and in 6% of controls. During this study, in one of an forestry workers the diagnosis of Lyme borreliosis as the occupational disease was clinically confirmed. High percent of anti-Borrelia burgdorferi antibodies found in people exposed to ticks may suggest that Borrelia burgdorferi is widespread in the forest environment of eastern Poland and that infection often has an occupational character. PMID- 9860822 TI - Divalent cations stabilize unstacked conformations of DNA and RNA by interacting with base pi systems. AB - Nucleic acid structure, stability, and reactivity are governed substantially by cations. We propose that magnesium and other biological inorganic ions unstack bases of DNA and RNA. This unstacking function of cations opposes their previously accepted role in stabilizing DNA and RNA duplexes and higher assemblies. We show that cations interact favorably with pi-systems of nucleic acid bases. These cation-pi interactions require access of cations or their first hydration shells to faces of nucleic acid bases. We observe that hydrated magnesium ions located in the major groove of B-DNA pull cytosine bases partially out from the helical stack, exposing pi-systems to positive charge. A series of critical cation-pi interactions contribute to the stability of the anticodon arm of yeast-tRNAphe, and to the magnesium core of the Tetrahymena group I intron P4 P6 domain. The structural consequences of divalent cation-pi interactions are clearly distinct from, and some cases in opposition to, cation-electron lone pair interactions. These observations of cation-pi interactions suggest a number of new mechanistic roles for cations in DNA bending, DNA-protein recognition, base flipping, RNA folding, and catalysis. PMID- 9860823 TI - X-ray absorption spectroscopy on layered photosystem II membrane particles suggests manganese-centered oxidation of the oxygen-evolving complex for the S0 S1, S1-S2, and S2-S3 transitions of the water oxidation cycle. AB - By application of microsecond light flashes the oxygen-evolving complex (OEC) was driven through its functional cycle, the S-state cycle. The S-state population distribution obtained by the application of n flashes (n = 0. 6) was determined by analysis of EPR spectra; Mn K-edge X-ray absorption spectra were collected. Taking into consideration the likely statistical error in the data and the variability stemming from the use of three different approaches for the determination of edge positions, we obtained an upshift of the edge position by 0.8-1.5, 0.5-0.9, and 0.6-1.3 eV for the S0-S1, S1-S2, and S2-S3 transitions, respectively, and a downshift by 2.3-3.1 eV for the S3-S0 transition. These results are highly suggestive of Mn oxidation state changes for all four S-state transitions. In the S0-state spectrum, a clearly resolved shoulder in the X-ray spectrum around 6555 eV points toward the presence of Mn(II). We propose that photosynthetic oxygen evolution involves cycling of the photosystem II manganese complex through four distinct oxidation states of this tetranuclear complex: Mn(II)-Mn(III)-Mn(IV)2 in the S0-state, Mn(III)2-Mn(IV)2 in the S1-state, Mn(III)1-Mn(IV)3 in the S2-state, and Mn(IV)4 in the S3-state. PMID- 9860824 TI - Approaching the transition state in the crystal structure of a phosphoribosyltransferase. AB - Hypoxanthine phosphoribosyltransferase (HPRT) salvages 6-oxopurine bases in the nucleotide metabolic pathway. The 1.8 A crystal structure of an asymmetric dimer of the HPRT from the protozoan parasite Trypanosoma cruzi was determined in a ternary complex with the primary substrate phosphoribosylpyrophosphate (PRPP) and an analogue of the substrate hypoxanthine, revealing both open and closed active site conformations. The ligands are positioned for in-line nucleophilic attack at the PRPP ribose C1' by two metal ions which straddle the pyrophosphate leaving group. The structure provides the first evidence for the involvement of two metal ions in the HPRT-catalyzed reaction, and structural details further suggest the mechanism may proceed via SN2-type chemistry. The closed conformation reveals the structural roles for invariant flexible loop residues Ser103 and Tyr104 and supports a role for the loop in the liberation of pyrophosphate. The pre transition state structure is valuable for understanding the enzyme mechanism, as well as providing a foundation for antiparasite drug design efforts against T. cruzi, which causes Chagas' disease in humans. Additionally, the structure illuminates the molecular basis of three inherited mutations in the human HPRT leading to Lesch-Nyhan syndrome (D193N) or gout (S103R or S109L), as the homologous residues in the trypanosomal enzyme contribute to the previously unrecognized Mg2+ ion binding site and to the formation of the closed flexible loop, respectively. PMID- 9860825 TI - Molecular basis for interprotein complex-dependent effects on the redox properties of amicyanin. AB - The quinoprotein methylamine dehydrogenase (MADH), type I copper protein amicyanin, and cytochrome c-551i form a complex within which interprotein electron transfer occurs. It was known that complex formation significantly lowered the oxidation-reduction midpoint potential (Em) value of amicyanin, which facilitated an otherwise thermodynamically unfavorable electron transfer to cytochrome c-551i. Structural, mutagenesis, and potentiometric studies have elucidated the basis for this complex-dependent change in redox properties. Positively charged amino acid residues on the surface of amicyanin are known to stabilize complex formation with MADH and influence the ionic strength dependence of complex formation via electrostatic interactions. Altering the charges of these residues by site-directed mutagenesis had no effect on the Em value of amicyanin, ruling out charge neutralization as the basis for the complex dependent changes in redox properties. The Em value of free amicyanin varies with pH and exhibits a pKa value for the reduced form of 7.5. The crystal structure of reduced amicyanin at pH 4.4 reveals that His95, which serves as a ligand for Cu2+, has rotated by 180 degrees about the Cbeta-Cgamma bond relative to its position in oxidized amicyanin and is no longer in the copper coordination sphere. At pH 7.7, the crystal structure of reduced amicyanin contains an approximately equal distribution of two active-site conformers. One is very similar to the structure of reduced amicyanin at pH 4.4, and the other is very similar to the structure of oxidized amicyanin at pH 4.8. Potentiometric analysis of amicyanin in complex with MADH indicates that its Em value is not pH-dependent from pH 6.5 to 8.5, and exhibits an Em value similar to that of free amicyanin at high pH. The structure of reduced amicyanin at pH 4.4, with His95 protonated and "flipped", was modeled into the structure of the complex of oxidized amicyanin with MADH. This showed that in the complex, the redox-linked pH-dependent rotation of His95 is hindered because it would cause an overlap of van der Waals' radii with residues of MADH. These results demonstrate that protein-protein interactions profoundly affect the redox properties of this type I copper protein by restricting a pH-dependent, redox-linked conformational change of one of the copper ligands. PMID- 9860826 TI - Functional characterization of the protease of human endogenous retrovirus, K10: can it complement HIV-1 protease? AB - To investigate the biochemical properties of the protease encoded by the human endogenous retrovirus, K10 (HERV-K), 213 amino acids of the 3'-end of the HERV-K protease (PR) open reading frame were expressed in Escherichia coli. Autocatalytic cleavage of the expressed polypeptide resulted in an 18.2 kDa protein which was shown to be proteolytically active against a fluorogenic peptide used as a substrate for HIV-1 protease. On the basis of sequence homology and molecular modeling, the 106 N-terminal amino acids of HERV-K PR were predicted to comprise a retroviral protease core domain. An 11.6 kDa protein corresponding to this region was expressed and shown to be a fully functional enzyme. The 11.6 kDa domain of HERV-K PR is unusually stable over a wide pH range, exhibits optimal catalytic activity between pH 4.0 and 5.0, and exists as a dimer at pH 7.0 with a Kd of 50 microM. Like HIV-1 PR, the HERV-K PR core domain is activated by high salt concentrations and processes HIV-1 matrix-capsid polyprotein at the authentic HIV-1 PR recognition site. However, both the 18.2 and 11.6 kDa forms of HERV-K PR were highly resistant to a number of clinically useful HIV-1 PR inhibitors, including ritonavir, indinavir, and saquinavir. This raises the possibility that HERV-K PR may complement HIV-1 PR during infection, and could have implications for protease inhibitor therapy and drug resistance. PMID- 9860827 TI - Reactivity of the human thioltransferase (glutaredoxin) C7S, C25S, C78S, C82S mutant and NMR solution structure of its glutathionyl mixed disulfide intermediate reflect catalytic specificity. AB - Human thioltransferase (TTase) is a 12 kDa thiol-disulfide oxidoreductase that appears to play a critical role in maintaining the redox environment of the cell. TTase acts as a potent and specific reducing agent for protein-S-S-glutathione mixed disulfides (protein-SSG) likely formed during oxidative stress or as redox intermediates in signal transduction pathways. Accordingly, the catalytic cycle of thioltransferase itself involves a covalent glutathionyl enzyme disulfide intermediate (TTase-C22-SSG). To understand the molecular basis of TTase specificity for the glutathione moiety, we engineered a quadruple Cys to Ser mutant of human TTase (C7S, C25S, C78S, and C82S) which retains only the active site cysteine residue (C22), and we solved its high-resolution NMR solution structure in the mixed disulfide intermediate with glutathione (QM-TTase-SSG). This mutant which cannot form a C22-S-S-C25 intramolecular disulfide displays the same catalytic efficiency (Vmax/KM) and specificity for glutathionyl mixed disulfide substrates as wild-type TTase, indicating that the Cys-25-SH moiety is not required for catalysis or glutathionyl specificity. The structure of human thioltransferase is characterized by a thioredoxin-like fold which comprises a four-stranded central beta-sheet flanked on each side by alpha-helices. The disulfide-adducted glutathione in the TTase-SSG complex has an extended conformation and is localized in a cleft near the protein surface encompassing the residues from helices-alpha2,alpha3, the active site loop, and the loop connecting helix-alpha3 and strand-beta3. Numerous van der Waals and electrostatic interactions between the protein and the glutathione moiety are identified as contributing to stabilization of the complex and confering the substrate specificity. Comparison of the human thioltransferase with other thiol disulfide oxidoreductases reveals structural and functional differences. PMID- 9860828 TI - IKP104-induced decay of tubulin: role of the A-ring binding site of colchicine. AB - Tubulin, the major subunit protein of microtubules, has a tendency to lose its ability to assemble or to interact with ligands in a time-dependent process known as decay. Decay involves the increase in exposure of sulfhydryl groups and hydrophobic areas. The antimitotic drug IKP104 [2-(4-fluorophenyl)-1-(2-chloro-3, 5-dimethoxyphenyl)-3-methyl-6-phenyl-4(1H)-pyridinone] accelerates the decay of tubulin [Luduena et al. (1995) Biochemistry 34, 15751-15759]. In the presence of colchicine, however, IKP104 stabilizes tubulin against decay. We have shown that the stability and the acceleration of the decay of tubulin are mediated respectively by the high- and low-affinity binding site(s) of IKP104 [Chaudhuri et al. (1998) J. Protein Chem. 17, 303-309]. To better understand the mechanism by which colchicine protects tubulin from IKP104-induced decay, we examined the effect of colchicine and its analogues on this process. We found that IKP104 unfolds tubulin in a process involving a specific domain where colchicine interacts, although the binding sites of these two drugs are distinctly different. 2-Methoxy-5-(2',3',4'-trimethoxyphenyl) tropolone (MTPT), the bicyclic analogue of colchicine that lacks the B-ring, can also protect tubulin from IKP104-induced decay. An A-ring analogue of colchicine, 3,4,5 trimethoxybenzaldehyde (TMB), can also stop IKP104-induced unfolding of tubulin significantly. Interestingly, the C-ring analogue of colchicine, tropolone methyl ether (TME), does not prevent this process. Our results thus suggest that neither the B-ring nor the C-ring binding regions of colchicine are involved in the IKP104-induced decay and that the A-ring binding site of colchicine on tubulin plays a crucial role in IKP104-induced decay. PMID- 9860829 TI - Conformational change rate-limits GTP hydrolysis: the mechanism of the ATP sulfurylase-GTPase. AB - The fluorescent GTP analogues 3'-O-(N-methylanthraniloyl)-2'-deoxyguanosine 5' (beta, gamma-imidotriphosphate) (mGMPPNP) and 3'-O-(N-methylanthraniloyl)-2' deoxy-GTP (mGTP) were used to demonstrate that an enzyme isomerization precedes and rate-limits beta,gamma-bond cleavage in the catalytic cycle of the ATP sulfurylase-GTPase, from E. coli K-12. The binding of mGMPPNP to the E.AMP.PPi complex of ATP sulfurylase is biphasic, indicating that an isomerization occurs in the binding reaction. The isomerization mechanism was assigned based on the results of the enzyme concentration dependence of the observed rate constants, kobs, for both phases of the binding reaction, and sequential-mixing, nucleotide release experiments. The isomerization occurs after, and is driven by, the addition of mGMPPNP. Values were determined for each of the rate constants associated with the two-step kinetic model used in the interpretation of the results. A comparison of the enzyme concentration dependence of kobs for the hydrolysis and binding reactions reveals that the rate constants for the corresponding steps of these two reactions are extremely similar. The virtually identical rate constants for isomerization and beta, gamma-bond scission strongly suggest that isomerization rate-limits bond breaking. The implications of these finding for GTPase/target interactions and the mechanism of energetic linkage in the ATP sulfurylase system are discussed. PMID- 9860830 TI - The family 1 beta-glucosidases from Pyrococcus furiosus and Agrobacterium faecalis share a common catalytic mechanism. AB - Comparisons of catalytic mechanisms have not previously been performed for homologous enzymes from hyperthermophilic and mesophilic sources. Here, the beta glucosidase from the hyperthermophilic archaeon Pyrococcus furiosus was recombinantly produced in Escherichia coli and shown to have biophyscial and biochemical properties identical to those of the wild-type enzyme. Moreover, the recombinant enzyme was subjected to a detailed kinetic investigation at 95 degreesC to compare its catalytic mechanism to that determined at 37 degreesC for the beta-glucosidase (abg) from the mesophilic bacterium, Agrobacterium faecalis [Kempton, J., and Withers, S. G. (1992) Biochemistry 31, 9961]. These enzymes have amino acid sequences that are 33% identical and have been classified as family 1 glycosyl hydrolases on the basis of amino acid sequence similarities. Both enzymes have similar broad specificities for both sugar and aglycone moieties and exhibit nearly identical pH dependences for their kinetic parameters with several different substrates. Bronsted plots were constructed for bgl at several temperatures using a series of aryl glucoside substrates. These plots were concave downward at all temperatures, indicating that bgl utilized a two step mechanism similar to that of abg and that the rate-limiting step in this mechanism did not change with temperature for any given aryl glucoside. The Bronsted coefficient for bgl at 95 degreesC (beta1g = -0.7) was identical to that for abg at 37 degreesC and implies that these enzymes utilize nearly identical transition states, at least in regard to charge accumulation on the departing glycosidic oxygen. In addition, a high correlation coefficient (rho = 0.97) for the linear free energy relationship between these two enzymes and similar inhibition constants for these two enzymes with several ground state and transition state analogue inhibitors further indicate that these enzymes stabilize similar transition states. The mechanistic similarities between these two enzymes are noteworthy in light of the large difference in their temperature optima. This suggests that, in the presumed evolution that occurred between the hyperthermophilic archaeal enzyme and the mesophilic bacterial enzyme, structural modifications must have been selected which maintained the integrity of the active site structure and, therefore, the specificity of transition state interactions, while adapting the overall protein structure to permit function at the appropriate temperature. PMID- 9860831 TI - Microsomal cytochrome P450 dependent oxidation of N-hydroxyguanidines, amidoximes, and ketoximes: mechanism of the oxidative cleavage of their C=N(OH) bond with formation of nitrogen oxides. AB - Oxidation by rat liver microsomes of 13 compounds involving a C=N(OH) function (including N-hydroxyguanidines, amidoximes, ketoximes, and aldoximes) was found to occur with the release of nitrogen oxides such as NO, NO2-, and NO3-. The greatest activities were observed with liver microsomes from dexamethasone treated rats (up to 8 nmol of NO2- nmol of P450(-)1 min-1). A detailed study of the microsomal oxidation of some of these compounds was performed. Oxidation of N (4-chlorophenyl)-N'-hydroxy-guanidine led to the formation of the corresponding urea and cyanamide in addition to NO, NO2-, and NO3-. Formation of all these products was dependent on NADPH, O2, and cytochromes P450. Oxidation of two arylamidoximes was found to occur with formation of the corresponding amides and nitriles in addition to nitrogen oxides. Oxidation of 4-(chlorophenyl)methyl ketone oxime gave the corresponding ketone and nitroalkane as well as NO, NO2-, and NO3-. These reactions were also dependent on cytochromes P450 and required NADPH and O2. Mechanistic experiments showed that microsomal oxidations of amidoximes to the corresponding nitriles and of ketoximes to the corresponding nitroalkanes are not inhibited by superoxide dismutase (SOD) and are performed by a cytochrome P450 active species, presumably the high-valent P450-iron-oxo complex. On the contrary, microsomal oxidation of N-hydroxyguanidines to the corresponding cyanamides was greatly inhibited by SOD and appeared to be mainly due to O2*- derived from the oxidase function of cytochromes P450. Similarly, microsomal oxidations of N-hydroxyguanidines and amidoximes to the corresponding ureas and amides were also found to be mainly performed by O2*-, as shown by the great inhibitory effect of SOD (70-100%) and the ability of the xanthine-xanthine oxidase system to give similar oxidation products. However, it is noteworthy that other species, such as the P450 Fe(II)-O2 complex, are also involved, to a minor extent, in the SOD-insensitive microsomal oxidative cleavages of compounds containing a C=N(OH) bond. Our results suggest a general mechanism for such oxidative cleavages of C=N(OH) bonds with formation of nitrogen oxides by cytochromes P450 and NO-synthases, with the involvement of O2*- and its Fe(III) complex [(FeIII-O2-) or (FeII-O2)] as main active species. PMID- 9860832 TI - Reassessment of acarbose as a transition state analogue inhibitor of cyclodextrin glycosyltransferase. AB - The binding of several different active site mutants of Bacillus circulans cyclodextrin glycosyltransferase to the inhibitor acarbose has been investigated through measurement of Ki values. The mutations represent several key amino acid positions, most of which are believed to play important roles in governing the product specificity of cyclodextrin glycosyltransferase. Michaelis-Menten parameters for the substrates alpha-maltotriosyl fluoride (alphaG3F) and alpha glucosyl fluoride (alphaGF) with each mutant have been determined by following the enzyme-catalyzed release of fluoride with an ion-selective fluoride electrode. In both cases, reasonable correlations are observed in logarithmic plots relating the Ki value for acarbose with each mutant and both kcat/Km and Km for the hydrolysis of either substrate by the corresponding mutants. This indicates that acarbose, as an inhibitor, is mimicking aspects of both the ground state and the transition state. A better correlation is observed for alphaGF (r = 0.98) than alphaG3F (r = 0.90), which can be explained in terms of the modes of binding of these substrates and acarbose. Re-refinement of the previously determined crystal structure of wild-type CGTase complexed with acarbose [Strokopytov, B., Penninga, D., Rozeboom, H. J., Kalk, K. H., Dijhuizen, L., and Dijkstra, B. W. (1995) Biochemistry 34, 2234-2240] reveals a binding mode consistent with the transition state analogue character of this inhibitor. PMID- 9860833 TI - Perturbation of the internal water chain in cytochrome f of oxygenic photosynthesis: loss of the concerted reduction of cytochromes f and b6. AB - The 1.96 A structure of turnip cytochrome f revealed a linear internal chain of H2O molecules with the oxygen atoms of the chain having occupancies and "B" factors comparable to those of neighboring atoms [Martinez et al. (1996) Protein Sci. 5, 1081-1092. ]. Four waters extend 11 A from the heme toward Lys66 on the cytochrome surface. All residues that contribute an atom to the 15 H-bonds of five internal H2O molecules are essentially conserved in 23 cytochrome sequences. With only Gln and Asn side chains involved in H-bonding, the water chain resembles a "proton wire". The function of the conserved H2O chain was tested through site-directed mutagenesis of these Asn and Gln residues. Four of the five conserved Asn/Gln residues were changed in six mutants generated in the green alga, Chlamydomonas reinhardtii. Except for the N168F mutant, all grew photosynthetically. Although the rates of oxidation of cyt f oxidation and of reduction of cyt b6 (5-6 ms in the wild type) were not significantly affected, the rates of cyt f reduction and generation of the slow electrochromic band shift (Deltapsis) were markedly decreased, the half-times increasing to as much as 38 and 18 ms, respectively. Thus, in these mutants, reduction of cyt b6 reduction clearly precedes that of cyt f. Retardation of Deltapsis in the absence of an observable change in the rate of cyt b6 reduction implied that the rate of H+ translocation decreased in the mutants, and electron transfer was concomitantly retarded, most likely between the ISP and cyt f. The following was concluded: (i) proton and electron transfer are coupled in reduction of cyt f, and the cyt f water chain functions in H+ transfer; (ii) reduction of the high- and low potential chains in the b6f complex is not concerted in the water chain mutants; and (iii) quinol deprotonation and electron transfer from reduced quinone are initiated by an early event, probably the movement of the ISP triggered by oxidation of cyt f. PMID- 9860834 TI - Influence of divalent cations on nucleotide exchange and ATPase activity of chloroplast coupling factor 1. AB - The ATPase activity of the catalytic part of ATP synthases is inhibited by free Mg2+, even though MgATP is the substrate. Here we show that the inhibition of the MgATPase activity of chloroplast coupling factor 1 deficient in its epsilon subunit (CF1-epsilon) by Mg2+ is complex. The hydrolysis of MgATP by CF1-epsilon that contains tightly bound ADP, but no bound Mg2+, is initially rapid and decreases within about 1 min to a steady-state rate. The bound MgADP content of CF1-epsilon was varied. The initial fast phase of MgATP hydrolysis is eliminated when the molar ratio of MgADP to CF1-epsilon approaches 2. Loosely bound Mg2+ also affects the initial kinetics of the enzyme that contains bound MgADP. At molar ratios of bound MgADP to enzyme in excess of 1, the initial ATPase activity was low and reached the steady state after about 30 s. Free Mg2+ in the assay mix also inhibited steady-state ATP hydrolysis by all forms of the enzyme. The results are consistent with a model in which two Mg2+ bind cooperatively, probably to the dissociable nucleotide-binding sites on CF1-epsilon. Thus, four different nucleotide-binding sites may be involved in the inhibition of the MgATPase activity of CF1-epsilon. Three of these sites are potentially catalytic, and the fourth may be regulatory. The exchange of bound trinitrophenyl-ADP induced by the addition of MgATP or CaATP was found to be fast enough for the site to be involved in catalysis. PMID- 9860835 TI - The hydrogen-bonding network of water molecules and the peptide backbone in the region connecting Asp83, Gly120, and Glu113 in bovine rhodopsin. AB - Difference Fourier transform infrared spectra were recorded between mutants of rhodopsin and their batho products. The pigments studied were single and combined mutants of intramembrane residues of bovine rhodopsin: Asp83, Glu113, Gly120, Gly121, and Glu122. Previous studies [Nagata, T., Terakita, A., Kandori, H., Kojima, D., Shichida, Y., and Maeda, A. (1997) Biochemistry 36, 6164-6170] showed that one of the water molecules which undergoes structural changes in this process forms hydrogen bonds with Glu113 and the Schiff base, and that another water molecule is linked to this structure through the peptide backbone. The present results show that this water molecule is located at the place that is affected by the replacements of Asp83 and Gly120 but only slightly by Gly121 and not at all by Glu122. Asp83 and Gly120 are close to each other, in view of the observations that the carboxylic C=O stretching vibration of Asp83 is affected by the G120A replacement and that each replacement affects the common peptide carbonyl groups. Our results suggest that these residues in the middle of helices B and C are linked-through a hydrogen-bonding network composed of water and the peptide backbone-with the region around Glu113. PMID- 9860836 TI - Characterization of truncated and glycosylation-deficient forms of the cation dependent mannose 6-phosphate receptor expressed in baculovirus-infected insect cells. AB - A soluble truncated form of the cation-dependent mannose 6-phosphate receptor (CD MPR) encoding only the extracytoplasmic region, Stop155, and a truncated glycosylation-deficient form of the CD-MPR, Asn81/Stop155, which has been modified to contain only one N-linked glycosylation site at position 81 instead of five, were purified from baculovirus-infected High Five insect cells. The glycosylated recombinant proteins were functional in ligand binding and acid dependent dissociation as assessed by pentamannosyl phosphate-agarose affinity chromatography. Gel filtration, sucrose gradients, and cross-linking experiments revealed that both Stop155 and Asn81/Stop155 are dimeric, demonstrating that the transmembrane and cytoplasmic region of the receptor as well as N-linked oligosaccharides at positions 31, 57, and 87 are not required for dimerization. The Kd of Stop155 and Asn81/Stop155 for the lysosomal enzyme, beta-glucuronidase, was 0.2 and 0.3 nM, respectively. These values are very similar to those reported for the full-length CD-MPR, demonstrating that the extracellular region of the CD MPR is sufficient for high-affinity binding and that oligosaccharides at positions 31, 57, and 87 do not influence ligand binding. PMID- 9860837 TI - Protein kinase C-dependent regulation of Na+/Ca2+ exchanger isoforms NCX1 and NCX3 does not require their direct phosphorylation. AB - We compared the phosphorylation-dependent regulation of three mammalian Na+/Ca2+ exchanger isoforms (NCX1-NCX3) expressed in CCL39 fibroblasts that have little endogenous activity. Na+i-dependent 45Ca2+ uptake into NCX1- or NCX3-expressing cells, but not that into NCX2-expressing cells, was significantly enhanced by phorbol 12-myristate 13-acetate (PMA) or platelet-derived growth factor-BB, which was abolished by pretreatment of cells with calphostin C or a prior long exposure to PMA. This suggests that NCX1 or NCX3, but not NCX2, is stimulated by a pathway involving protein kinase C (PKC). Immunoprecipitation experiments using [32P]orthophosphate-labeled cells revealed that both NCX2 and NCX3 proteins were phosphorylated to a much lesser extent than the NCX1 protein in unstimulated cells and that the extent of phosphorylation was not increased by treatment with PKC activators, although NCX1 phosphorylation was enhanced significantly. Using site-directed mutagenesis, we identified three phosphorylation sites in the NCX1 protein in the PMA-stimulated cells to be Ser-249, Ser-250, and Ser-357 with Ser 250 being predominantly phosphorylated. We found that the NCX1 mutant with these serine residues substituted with alanine still maintained a normal response to PMA. In contrast, the NCX1 or NCX3 mutant, with the large central cytoplasmic loop deleted, lost the responsiveness to PMA. These results suggest that the PKC dependent regulation of NCX1 or NCX3 requires the central cytoplasmic loop but does not require the direct phosphorylation of the exchanger. PMID- 9860838 TI - Three residues predicted by molecular modeling to interact with the purine moiety alter ligand binding and channel gating in cyclic nucleotide-gated channels. AB - Cytoplasmic cAMP and cGMP are soluble cellular messengers that directly activate cyclic nucleotide-gated (CNG) channels. These channels mediate sensory transduction in photoreceptors and olfactory neurons. The closely related CNG channels in these cell types have different nucleotide activation profiles, and we have investigated the molecular basis of their nucleotide selectivity properties. Previously, we predicted that the purine moiety of the nucleotide interacts with residues F533, K596, and D604 (bovine rod alpha CNG channel subunit sequences) of the nucleotide binding domain. In this study, we replaced these three residues with the corresponding residues of the bovine olfactory CNG channel. Mutations at each position altered the nucleotide activation of the rod CNG channels. In a mutant where K596 was replaced with arginine, cAMP-activated currents were enhanced 8-12-fold, suggesting that residue 596 influences channel gating. Thermodynamic cycle analysis of the data showed that (1) the residues are energetically coupled and (2) energetic coupling exists between the potentiating effects of Ni2+ and the replacement of F533 with tyrosine. These data suggest that changes in one of the residues alter the purine contacts with the other residues and that F533 communicates with the C-linker region of the channel involved in Ni2+ potentiation. PMID- 9860840 TI - Restoration of the activity of active-site mutants of the hyperthermophilic beta glycosidase from Sulfolobus solfataricus: dependence of the mechanism on the action of external nucleophiles. AB - The beta-glycosidase from the hyperthermophilic Archaeon Sulfolobus solfataricus hydrolyzes beta-glycosides following a retaining mechanism based upon the action of two amino acids: Glu387, which acts as the nucleophile of the reaction, and Glu206, which acts as the general acid/base catalyst. The activities of inactive mutants of the catalytic nucleophile Glu387Ala/Gly were restored by externally added nucleophiles. Sodium azide and sodium formate were used as external nucleophiles and the products of their reaction were characterized. Glu387Ala/Gly mutants were reactivated with 2, 4-DNP-beta-Glc substrate and the Glu387Gly mutant showed recovered activity, with the same nucleophiles, also on 2-NP-beta Glc. The reaction catalyzed by the Glu387Gly mutant proceeded differently depending on the type of externally added nucleophile. Sodium azide restored the catalytic activity of the mutant by attacking the alpha-side of the anomeric carbon of the substrates, thereby yielding an inverting glycosidase. Sodium formate promoted the opposite behavior (retaining) in the mutant, producing 3-O beta-linked disaccharide derivative of the substrates. A possible role of sodium formate as a biomimicking agent in replacing the natural nucleophile Glu387 is also discussed. PMID- 9860839 TI - Calnexin and BiP interact with acid phosphatase independently of glucose trimming and reglucosylation in Schizosaccharomyces pombe. AB - The association of newly synthesized glycoproteins with the ER molecular chaperones calnexin and immunoglobulin binding protein (BiP) has been well documented in a variety of higher eukaryotes. Here we report that Cnx1p, the calnexin homologue in Schizosaccharomyces pombe, associates with newly synthesized molecules of the secreted glycoprotein acid phosphatase. Unlike ligand binding to mammalian calnexin, glucose trimming and reglucosylation of acid phosphatase by UDP-Glc:glycoprotein glucosyltransferase were shown to be dispensable for its binding to Cnx1p. Thus, despite the essentiality of Cnx1p for S. pombe viability, the glucose trimming and reglucosylation cycle does not appear to be required for protein folding in the fission yeast. The association of core-glycosylated acid phosphatase with Cnx1p after exposure of cells to heat shock or to DTT was shown to be reversible. However, Cnx1p stably associated with unglycosylated acid phosphatase after treatment with the core-glycosylation inhibitor tunicamycin. BiP was found to coprecipitate with Cnx1p, under normal and stress conditions, and following inhibition of protein synthesis by cycloheximide. We postulate that Cnx1p and BiP are part of a complex that is involved in the folding of both core-glycosylated trimmed ligands and unglycosylated proteins. PMID- 9860841 TI - Mechanism of the apparent cooperativity in the interaction of protein kinase C with phosphatidylserine. AB - Protein kinase C displays high apparent cooperativity in its activation by phosphatidylserine. This contribution uses a novel approach to address the physical basis for this apparent cooperativity. We examine the binding of protein kinase C betaII to large unilamellar vesicles as a function of increasing mole fraction phosphatidylserine and as a function of increasing total lipid concentrations. Binding data are subjected to an analysis, described in the Appendix, that allows calculation of the fractional saturation of phosphatidylserine binding sites with this ligand. This analysis reveals that (1) protein kinase C betaII binds approximately eight phosphatidylserine molecules and (2) the binding of each lipid is not cooperative. Rather, the apparent cooperativity observed in protein kinase C's interaction with multiple phosphatidylserine molecules arises from effects specific to the interaction of a multivalent macromolecule with multiple membrane-associated ligands. Nor does diacylglycerol, which has been previously shown to dramatically increase protein kinase C's affinity for phosphatidylserine-containing membranes, induce cooperativity. Thus, protein kinase C binds multiple phosphatidylserine molecules in the absence of interaction between potential binding sites. The method presented for determining the stoichiometry and cooperativity in the interaction of protein kinase C with phosphatidylserine is applicable to any multivalent molecule binding to monovalent ligands incorporated into lipid membranes. PMID- 9860842 TI - Thiol-mediated disassembly and reassembly of [2Fe-2S] clusters in the redox regulated transcription factor SoxR. AB - SoxR, a transcription factor containing [2Fe-2S] clusters, governs the cellular response to oxidative stress in Escherichia coli. The oxidation state of the iron sulfur clusters regulates the SoxR transcriptional activity. When the reduced iron-sulfur clusters become oxidized ([2Fe-2S]2+ state), SoxR is activated to stimulate transcription of the soxS gene, whose product in turn switches on a group of genes encoding various proteins that defend against oxidative stress and antibiotics. A previous study showed that the oxidized [2Fe-2S] clusters of SoxR are destroyed by a free-radical-dependent process in vitro during aerobic exposure to the biological thiol glutathione. Here, we show that different thiols have differing effects on the SoxR [2Fe-2S] clusters. Like reduced glutathione, N acetyl-L-cysteine, L-cysteine methyl ester, and L-cysteine ethyl ester disrupted the SoxR [2Fe-2S] clusters in aerobic solution. This disruption was blocked by L cysteine, which was effective at concentrations 100-fold lower (1-10 microM) than the disrupting thiols (1 mM). In view of a previous observation that superoxide dismutase and catalase block the disruption process, this result suggests that L cysteine may quench reactive SoxR or thiol intermediates involved in the cluster disruption reaction, the detailed mechanism of which remains unknown. In contrast, bifunctional thiols such as dithiothreitol or dithioerythritol promoted the aerobic assembly of the functional [2Fe-2S] clusters into apo-SoxR in the presence of Fe2+ and inorganic sulfide. The dithiol protein thioredoxin-A of E. coli acted catalytically in vitro in the presence of thioredoxin reductase and NADPH to promote [2Fe-2S] cluster assembly into apo-SoxR. The regulatory activity of SoxR in vivo, assessed by monitoring the paraquat-mediated induction of a soxS'::lacZ reporter fusion, was significantly lower in a strain lacking both thioredoxin-A and glutathione reductase, which maintains reduced glutaredoxins. Thus, cellular monothiols and dithiol proteins may contribute to SoxR regulation by affecting the disassembly and reassembly of the [2Fe-2S] clusters. PMID- 9860844 TI - Influence of docosahexaenoic acid and cholesterol on lateral lipid organization in phospholipid mixtures. AB - We investigated lateral lipid organization in membranes with a lipid composition relevant to neural and retinal membranes [phosphatidylcholine (PC)/phosphatidylethanolamine (PE)/phosphatidylserine (PS)/cholesterol, 4/4/1/1, mol/mol/mol/mol]. The mixed-chain phospholipids contained saturated stearic acid (18:0) in the sn-1 position and the monounsaturated oleic acid (18:1) or polyunsaturated docosahexaenoic acid (22:6) in sn-2. Lateral lipid organization was evaluated by 2H NMR order parameter measurements on stearic acid of all individual types of phospholipids in the mixture and, through a novel approach, two-dimensional NOESY 1H NMR spectroscopy with magic angle spinning (MAS). The docosahexaenoic acid chain order was evaluated from 1H NMR chain signal MAS sideband intensities. Averaged over all lipids, the cholesterol-induced increase in sn-1 chain order is 2-fold larger in monounsaturated than in polyunsaturated lipids, and the order of both saturated and polyunsaturated hydrocarbon chains increases. Addition of cholesterol increases lipid order in the sequence 18:0 18:1 PE > 18:0-18:1 PC > 18:0-18:1 PS for the monounsaturated and 18:0-22:6 PC >> 18:0-22:6 PE > 18:0-22:6 PS for polyunsaturated mixtures. The variation of order parameters between lipid species suggests that cholesterol induces the formation of lipid microdomains with a headgroup and chain unsaturation-dependent lipid composition. The preferential interaction between cholesterol and polyunsaturated 18:0-22:6 PC, followed by 18:0-22:6 PE and 18:0-22:6 PS, was confirmed by 1H MAS NOESY cross-relaxation rate differences. Furthermore, cholesterol preferentially associates with saturated chains in mixed-chain lipids reflected by higher saturated chain-to-cholesterol cross-relaxation rates. We propose that cholesterol forms PC-enriched microdomains in the polyunsaturated 18:0-22:6 PC/18:0-22:6 PE/18:0-22:6 PS/cholesterol membranes in which the saturated sn-1 chains are preferentially oriented toward the cholesterol molecules. PMID- 9860843 TI - Estrogen receptor alpha requires no accessory factors for high-affinity binding to a consensus response element. AB - Estrogen receptor (ER) alpha is commonly thought to bind to a consensus estrogen response element (ERE) as a homodimer, but previous experiments have not ruled out the presence of other proteins in the ERalpha/ERE complex. To characterize this interaction in more detail, we overexpressed mouse (m) ERalpha in a baculovirus system, using the selective advantage of the apoptosis inhibitor p35. Recombinant mERalpha possesses the predicted molecular weight and binds 17beta estradiol and an oligonucleotide containing a consensus vitellogenin ERE with high affinity. Over a wide concentration range of mERalpha protein (0.1-50 nM), only one complex was detected between mERalpha and vitellogenin ERE in gel shift assays. The ratio of E2:vitellogenin ERE bound by mERalpha was close to 2:1, and each complex contained only one ERE. The molecular weight of the complex was determined to be 160 000, very close to that predicted for two mERalpha proteins and one ERE oligonucleotide, therefore providing strong evidence that no other proteins were present. Recombinant mERalpha was purified such that it was the only protein observable by silver stain. Purified mERalpha and mERalpha in a nuclear extract behaved identically in Ferguson analysis, providing more evidence that only mERalpha was binding to the ERE. Purified mERalpha bound vitellogenin ERE with high affinity (Kd = 0. 92 +/- 0.20 nM), indicating that no other proteins are necessary for high-affinity mERalpha interaction with a consensus ERE. To determine whether ERalpha in an estrogen-responsive mammalian tissue behaves the same as the overexpressed mERalpha, we tested rat uterine cytosol by Ferguson analysis. ERalpha in rat uterine cytosol behaved identically to overexpressed mERalpha, suggesting that ERalpha in the uterine extract also binds to DNA predominantly as a homodimer with no additional proteins. PMID- 9860845 TI - Two stable unfolding intermediates of the disease-causing L68Q variant of human cystatin C. AB - In hereditary cystatin C amyloid angiopathy (HCCAA), presence of the Leu68 --> Gln substitution in cystatin C is coupled to a decreased concentration of this major cysteine proteinase inhibitor in cerebrospinal fluid and leads to its amyloid deposition in the brain. We established a high-yield expression system for L68Q cystatin C in Escherichia coli resulting in inclusion body accumulation at a level of 40% of the total cellular protein. Refolding of protein from purified inclusion bodies yielded a pure, almost completely monomeric and active inhibitor. CD and NMR spectroscopy demonstrated that so produced L68Q cystatin C is folded, conformationally homogeneous, and structurally very similar to wild type cystatin C. Incubation at pH 7.0-5.5 caused the cystatin C variant to dimerize rapidly. The molecular form present at pH 6.0 displayed a slightly increased amount of hydrophobic parts on the surface as measured by 1 anilinonaphthalene-8-sulfonic acid (ANS) binding. NMR results showed that the dimer has a structure similar to that of the wild-type cystatin C dimer formed as a result of slight denaturation. Under more acidic conditions, at pH 4.5, another stable unfolding intermediate of L68Q cystatin C was identified. This molecular form exists in a monomeric state, is characterized by changes in secondary structure according to far UV CD spectroscopy, and shows an altered ANS binding resembling that of a molten globule state. The acidic pH also caused an almost complete monomerization of preformed dimers. The state of denaturation of L68Q cystatin C in vivo is thus a critical factor for the concentration of active cysteine proteinase inhibitor in cerebrospinal fluid and likely also for the development of amyloidosis, in HCCAA patients. PMID- 9860846 TI - Helix-stabilizing nonpolar interactions between tyrosine and leucine in aqueous and TFE solutions: 2D-1H NMR and CD studies in alanine-lysine peptides. AB - Interactions between side chains spaced (i,i + 3) and (i,i + 4) may explain the context dependence of helix propensities observed in different systems. Nonpolar residues with these spacings occur frequently in protein helices and stabilize isolated peptide helices. Here (i,i + 3) and (i,i + 4) nonpolar interactions between Tyr and Leu in different solution conditions are studied in detail in alanine-based peptides using 2D 1H NMR and CD spectroscopy. Helix contents analyzed using current models for helix-coil transitions yield interaction energies which demonstrate significant helix stabilization in aqueous 1 M NaCl solutions by Tyr-Leu or Leu-Tyr pairs when spaced (i,i + 4) and, to a smaller extent, when spaced (i,i + 3), comparable to those estimated for other residue pairs. The interactions persist in solutions containing TFE, a helix-stabilizing solvent believed to diminish hydrophobic interactions, but not in helix destabilizing 6 M urea. 1H NMR resonances for all peptides and solution conditions except in 6 M urea were completely assigned. NMR data indicate that the N-terminal residues are more helical and that the N-acetyl group participates in helix formation. The two (i,i + 4) spaced pairs show the same pattern of NOE cross-peaks between the Tyr and Leu side chains, as do the two (i,i + 3) pairs in 1 M NaCl as well in TFE solutions, and correspond well with that expected for the specific Tyr-Leu pair with side-chain contacts in protein helices. PMID- 9860847 TI - Multiple states of beta-sheet peptide protegrin in lipid bilayers. AB - Protegrin-1 (PG-1), a beta-sheet antimicrobial peptide, was studied in aligned lipid bilayers by oriented circular dichroism (OCD). All of its spectra measured in a variety of lipid compositions were linear superpositions of two primary basis spectra, indicating that PG-1 existed in two different states in membranes. We designated these as state S and state I. The state assumed by PG-1 was strongly influenced by lipid composition, peptide concentration, and hydration condition. We have previously reported that the helical peptides, alamethicin and magainin, also exhibit two distinct OCD basis spectra-one corresponding to surface adsorption with the helix parallel to the bilayer and the other with perpendicular transbilayer insertion. States S and I of PG-1 may correspond to the surface state and the insertion state of alamethicin, since they show a similar dependence on lipid composition, peptide concentration, and hydration condition. Nonoriented CD spectra obtained from vesicle, micelle, and solution preparations are not linear superpositions of the basis spectra of the states S and I. This indicates that a molecular orientation change alone is insufficient to describe the S left and right arrow I transition. Rather, a more complicated process is taking place, perhaps involving a change in the hydrogen bonding pattern of the backbone. Although the structural basis of the OCD spectra remains to be determined, the discovery of two distinct states can provide information about dynamic changes of PG-1 in membranelike environments, properties undoubtedly related to its antimicrobial and cytotoxic effects. PMID- 9860848 TI - Influence of herbicide binding on the redox potential of the quinone acceptor in photosystem II: relevance to photodamage and phytotoxicity. AB - Here we show that herbicide binding influences the redox potential (Em) of the plastoquinone QA/QA- redox couple in Photosystem II (PSII). Phenolic herbicides lower the Em by approximately 45 mV, while DCMU raises the Em by 50 mV. These shifts are reflected in changes in the peak temperature of thermoluminescence bands arising from the recombination of charge pairs involving QA-. The herbicide induced changes in the Em of QA/QA- correlate with earlier work showing that phenolic herbicides increase the sensitivity of PSII to light, while DCMU protects against photodamage. This correlation is explained in terms of the following hypothesis which is based on reactions occurring in the bacterial reaction center. The back-reaction pathway for P680+QA- is assumed to be modulated by the free-energy gap between the P680+QA- and the P680+Ph- radical pairs. When this gap is small (i.e., when the Em of QA/QA- is lowered), a true back-reaction is favored in which P680+Ph- is formed, a state which decays forming a significant yield of P680 triplet. This triplet state of chlorophyll reacts with oxygen, forming singlet oxygen, a species likely to be responsible for photodamage. When the free-energy gap is increased (i.e., when the Em of QA/QA- is raised), the yield of the P680+Ph- is diminished and a greater proportion of the P680+QA- radical pair decays by an alternative, less damaging, route. We propose that at least some of the phytotoxic properties of phenolic herbicides may be explained by the fact that they render PSII ultrasensitive to light due to this mechanism. PMID- 9860849 TI - Nitrogenase of Azotobacter vinelandii: kinetic analysis of the Fe protein redox cycle. AB - Nitrogenase consists of two metalloproteins (Fe protein and MoFe protein) which are assumed to associate and dissociate to transfer a single electron to the substrates. This cycle, called the Fe protein cycle, is driven by MgATP hydrolysis and is repeated until the substrates are completely reduced. The rate limiting step of the cycle, and substrate reduction, is suggested to be the dissociation of the Fe protein-MoFe protein complex which is obligatory for the reduction of the Fe protein [Thorneley, R. N. F., and Lowe, D. J. (1983) Biochem. J. 215, 393-403]. This hypothesis is based on experiments with dithionite as the reductant. We also tested besides dithionite flavodoxin hydroquinone, a physiological reductant. Two models could describe the experimental data of the reduction by dithionite. The first model, with no reduction of Fe protein bound to MoFe protein, predicts a rate of dissociation of the protein complex of 8.1 s 1. This rate is too high to be the rate-limiting step of the Fe protein cycle (kobs = 3.0 s-1). The second model, with reduction of the Fe protein in the nitrogenase complex, predicts a rate of dissociation of the protein complex of 2.3 s-1, which in combination with reduction of the nitrogenase complex can account for the observed turnover rate of the Fe protein cycle. When flavodoxin hydroquinone (155 microM) was the reductant, the rate of reduction of oxidized Fe protein in the nitrogenase complex (kobs approximately 400 s-1) was 100 times faster than the turnover rate of the cycle with flavodoxin as the reductant (4 s 1). Pre-steady-state electron uptake experiments from flavodoxin hydroquinone indicate that before and after reduction of the nitrogenase complex relative slow reactions take place, which limits the rate of the Fe protein cycle. These results are discussed in the context of the kinetic models of the Fe protein cycle of nitrogenase. PMID- 9860850 TI - A thermal broadening study of the antenna chlorophylls in PSI-200, LHCI, and PSI core. AB - The intact photosystem I of maize containing its full antenna complement (PSI 200) has been purified and fractionated into the core and outer antenna (LHCI) components. It is demonstrated by absorption and fluorescence spectroscopy that at least 80% of the long wavelength absorbing antenna pigments (red forms) are located in LHCI. Absorption spectra in the Qy region of all three preparations were measured between 72 and 300 K and subjected to a thermal broadening analysis. Data are interpreted in the linear electron-phonon coupling assumption, and the average optical reorganization energy (Snum) for the bulk pigment band and the red absorption tail determined. A marked asymmetry in Snum values across the absorption band is demonstrated. The bulk pigments in all three preparations have rather low values, in the range of 15-25 cm-1, suggesting that Stokes shifts for the absorption forms are in the 1. 5-3 nm range. On the other hand the red forms have markedly greater reorganization energies. While a direct thermal analysis of the red tail indicates minimum Snum values of around 60 cm-1, when the contribution of the red tail of the bulk pigments is corrected for in LHCI, the more reliable value of 110 cm-1 is obtained. These high Snum values for the red pigment forms suggest that they have unusually wide homogeneously broadened absorption bands and large Stokes shifts (6-11 nm). PMID- 9860851 TI - Coagulation factor X-binding protein from Deinagkistrodon acutus venom is a Gla domain-binding protein. AB - Factor IX/factor X-binding protein (IX/X-bp) is an anticoagulant isolated from the venom of Trimeresurus flavoviridis (habu snake) and binds predominantly to factor IX. In this study, we isolated IX/X-bp-like proteins from the venom of Deinagkistrodon acutus (hundred pace snake) with binding characteristics different from those of IX/X-bp. The complete amino acid sequence and binding characteristics of the main anticoagulant protein, named X-bp, were investigated. The concentrations of X-bp at half-maximal binding to solid-phase factors X and IX were 0.4 and 3 nM, respectively. The binding of X-bp to solid-phase factor X was inhibited by 50% by 6- and 9-fold excess concentrations of factor X and Gla domain (GD) peptide 1-44, respectively, but was not influenced by GD peptide 1-41 and Gla domainless factor X. X-bp bound two Ca2+ ions per molecule with Kd values of 16 +/- 0.7 (mean +/- SE, n = 6) and 103 +/- 10 microM. X-bp was a heterodimer of C-type lectin-like subunits. The 16 kDa chain (A chain) consisted of 129 amino acid residues and was 68% identical to the sequence of the A chain of IX/X-bp. The 15 kDa chain (B chain) consisted of 123 amino acid residues and was 87% identical to IX/X-bp. Three-dimensional model construction from the known fold of IX/X-bp showed that amino acid residues different from those of IX/X-bp are mostly on the molecular surface. Some of these are concentrated on a part of the concave surface which is considered to be the coagulation factor-binding site, presumably acting as a discriminator for ligand binding. These results indicated that X-bp isolated from D. acutus venom was a GD-binding protein, and the C terminal region of GD peptide was critical for folding of the peptide. PMID- 9860852 TI - Kinetic isomers of a class II MHC-peptide complex. AB - Class II major histocompatibility (MHC) molecules bind fragments of antigens and present them to T cells. The triggering of the T-cell receptor (TCR) of CD4(+) T helper cells by these protein-peptide complexes is a key event in the generation of a cellular immune response. In the context of this interaction, it is generally assumed that class II MHC-peptide complexes adopt a single recognition structure at the cell surface. On the other hand, kinetic analysis has revealed that a number of class II MHC-peptide complexes show biphasic dissociation kinetics, indicating the presence of multiple kinetic isomers. Here, we demonstrate that a water-soluble version of the murine class II MHC molecule I-Ek complexed with an antigenic peptide derived from pigeon cytochrome c (PCC) displays monophasic as well as biphasic dissociation kinetics. While a simple monophasic dissociation curve was obtained at neutral pH, the complex showed biphasic dissociation behavior at acidic pH. This shift was independent of the ionic strength of the solution. Moreover, the short-lived isomer could be regenerated from a pool of kinetically homogeneous long-lived complexes. This demonstrates that the isomers interconvert and exist in a pH-sensitive equilibrium. Altering the peptide residue of PCC that occupies the P6 pocket of I Ek results in a class II MHC-peptide complex that shows only monophasic dissociation, indicating that the glutamine at this position plays a key role in the kinetic heterogeneity of the complex. PMID- 9860853 TI - The magnitude of the allosteric conformational transition of aspartate transcarbamylase is altered by mutations. AB - Global conformational transitions are of central functional importance for many enzymes and binding proteins. It is not known, however, how much variability can exist in such structural-functional linkages. We have characterized the global magnitude of the T to R conformational transition of Escherichia coli aspartate transcarbamylase (ATCase) by measuring (1) hydration changes using osmotic stress and (2) hydrodynamic changes using high-precision analytical gel chromatography. We find that specific mutations can alter the structural magnitude of the enzyme's conformational transition without abolishing allostery, suggesting that some degree of plasticity exists in the conformational component of allostery. PMID- 9860854 TI - Coulombic effects of remote subsites on the active site of ribonuclease A. AB - The active-site cleft of bovine pancreatic ribonuclease A (RNase A) is lined with cationic residues that interact with a bound nucleic acid. Those residues interacting with the phosphoryl groups comprise the P0, P1, and P2 subsites, with the scissile P-O5' bond residing in the P1 subsite. Coulombic interactions between the P0 and P2 subsites and phosphoryl groups of the substrate were characterized previously [Fisher, B. M., Ha, J.-H., and Raines, R. T. (1998) Biochemistry 37, 12121-12132]. Here, the interactions between these subsites and the active-site residues His12 and His119 are described in detail. A protein variant in which the cationic residues in these subsites (Lys66 in the P0 subsite and Lys7 and Arg10 in the P2 subsite) were replaced with alanine was crystallized, both free and with bound 3'-uridine monophosphate (3'-UMP). Structures of K7A/R10A/K66A RNase A and the K7A/R10A/K66A RNase A.3'-UMP complex were determined by X-ray diffraction analysis to resolutions of 2.0 and 2.1 A, respectively. There is little observable change between these structures and that of wild-type RNase A, either free or with bound 3'-cytidine monophosphate. K7A/R10A/K66A RNase A was evaluated for its ability to cleave UpA, a dinucleotide substrate that does not span the P0 or the P2 subsites. In comparison to the wild type enzyme, the value of kcat was decreased by 5-fold and that of kcat/Km was decreased 10-fold, suggesting that these remote subsites interact with the active site. These interactions were characterized by determining the pKa values of His12 and His119 at 0.018 and 0.142 M Na+, both in wild-type RNase A and the K7A/R10A/K66A variant. The side chains of Lys7, Arg10, and Lys66 depress the pKa values of these histidine residues, and this depression is sensitive to the salt concentration. In addition, the P0 and P2 subsites influence the interaction of His12 and His119 with each other, as demonstrated by changes in the cooperativity that gives rise to microscopic pKa values. Finally, the affinity of 3'-UMP for wild-type RNase A and the K7A/R10A/K66A variant at 0.018 and 0.142 M Na+ was determined by isothermal titration calorimetry. 3'-UMP binds to the variant protein with 5-fold weaker affinity at 0.018 M Na+ and 3-fold weaker affinity at 0.142 M Na+ than it binds to wild-type RNase A. Together these data demonstrate that long-range Coulombic interactions are an important feature in catalysis by RNase A. PMID- 9860855 TI - Engineering of S2 site of aqualysin I; alteration of P2 specificity by excluding P2 side chain. AB - Gly101, one of the conserved amino acid residues which was expected to be comprised in half-sphere-shaped S2 site small pocket of aqualysin I, a microbial thermophilic alkaline serine protease, was replaced by alanine, valine, or leucine to alterate the P2 specificity of the enzyme by excluding bulky P2 side chain of the substrate. By the mutation of G101A, the catalytic efficiencies of the enzyme for bulky amino acid residues in P2 site such as valine and leucine drastically decreased by excluding the P2 side chain. By the mutation of G101V, even the side chain of the methyl group of the alanine and the side chain of proline were excluded, while the catalytic efficiency toward glycine residue was retained. The enzyme was altered to be glycine preferable. The mutation of G101L reduced catalytic efficiencies for any substrate including glycine which is corresponding to the main chain of the peptide substrate. The strategies we have adopted in this paper are applicable to all subtilisin-related enzymes. PMID- 9860856 TI - Core mutations that promote the calcium-induced allosteric transition of bovine recoverin. AB - Recoverin is a small calcium binding protein involved in regulation of the phototransduction cascade in retinal rod cells. It functions as a calcium sensor by undergoing a cooperative, ligand-dependent conformational change, resulting in the extrusion of the N-terminal myristoyl group from a hydrophobic pocket. To test the role of certain core residues in tuning this allosteric switch, we have made and characterized two mutants: W31K, which replaces Trp31 with Lys; and a double mutant, I52A/Y53A, in which Ile52 and Tyr53 are both replaced by Ala. These mutations decrease the hydrophobicity of the myristoyl binding pocket. They are thus expected to make sequestering of the myristoyl group less favorable and destabilize the Ca2+-free state. As predicted, the myristoylated forms of the mutants exhibit increased affinity for Ca2+, whether monitored by equilibrium binding of 45Ca2+ (Kd = 17.2, 7.9, and 8.1 microM for wild type, W31K, and I52A/Y53A, respectively) or by the change in tryptophan fluorescence associated with the conformational change (Kd = 17.9, 3.6, and 4.4 microM for wild type, W31K, and I52A/Y53A, respectively). The mutants also exhibit decreased cooperativity of binding (Hill coefficient = 1.2 and 1.0 for W31K and I52A/Y53A vs 1. 4 for wild type). Binding of the mutant proteins to rod outer segment membranes occurs at lower Ca2+ concentrations compared to wild-type protein (K1/2 = 5.6, 2.2, and 1.0 microM for wild type, W31K, and I52A/Y53A, respectively). The unmyristoylated forms of the mutants exhibit biphasic Ca2+ binding curves, nearly identical to that observed for wild type. The binding data for the two mutants can be explained by a concerted allosteric model in which the mutations affect only the equilibrium constant L between the two allosteric forms, T (the Ca2+ free form) and R (the Ca2+-bound form), without affecting the intrinsic binding constants for the two Ca2+ sites. Two-dimensional NMR spectra of the Ca2+-free forms of the mutants have been compared to the wild-type spectrum, whose peaks have been assigned to specific residues (1). Many resonances assigned to residues in the C-terminal domain (residues 100-202) in the wild-type spectrum are identical in the mutant spectra, suggesting that the backbone structure of the C terminal domain is probably unchanged in both mutants. The N-terminal domain, in which both mutations are located, reveals in each case numerous changes of undetermined spatial extent. PMID- 9860857 TI - The P histo-blood group-related glycosphingolipid sialosyl galactosyl globoside as a preferred binding receptor for uropathogenic Escherichia coli: isolation and structural characterization from human kidney. AB - The P histo-blood group-related glycosphingolipid, sialosyl galactosyl globoside (SGG), has recently been implicated as a preferred binding receptor for uropathogenic Escherichia coli [Stapleton, A. E., Stroud, M. R., Hakomori, S., and Stamm, W. E. (1998) Infect. Immun. 66, 3856-3861]. We report here the purification and complete structural characterization of SGG from normal human kidney. Using metabolically [35S]-labeled E. coli as a probe, a monosialylated glycosphingolipid was isolated to homogeneity. The glycosphingolipid was purified by a combination of high-performance liquid chromatography and preparative high performance thin-layer chromatography and its structure unambiguously elucidated by 1H NMR, electrospray ionization mass spectrometry, and methylation analysis. Its primary structure was shown to be identical to a previously characterized, developmentally regulated, globo-series glycolipid thought to be unique to human teratocarcinoma. The significance of this structure as a unique receptor in human kidney for uropathogenic E. coli and its role in the pathogenesis of urinary tract infections are discussed. PMID- 9860858 TI - S100A1 utilizes different mechanisms for interacting with calcium-dependent and calcium-independent target proteins. AB - While previous studies have identified target proteins that interact with S100A1 in a calcium-dependent manner as well as target proteins that interact in a calcium-independent manner, the molecular mechanisms of S100A1-target protein interaction have not been elucidated. In this study, point and deletion mutants of S100A1 were used to investigate the contribution of carboxyl terminal amino acids to S100A1 interaction with calcium-dependent and calcium-independent target proteins. First, a recombinant rat S100A1 protein (recS100A1) expressed in bacteria exhibited physical and chemical properties indistinguishable from native S100A1. Next, proteins lacking the carboxyl-terminal nine residues of recS100A1 (Delta85-93), or containing alanine substitutions at Phe 88 (F88A), Phe 89 (F89A), or Trp 90 (W90A), both Phe 88 and Phe 89 (F88/89A), or all three aromatic residues (F88/89A-W90A) were recombinantly expressed. Like recS100A1, F88A, F89A, and W90A proteins interacted with phenyl-Sepharose in a calcium-dependent manner. However, the Delta85-93 protein did not interact with phenyl-Sepharose, indicating that a phenyl-Sepharose-binding region (PSBR) of recS100A1 had been disrupted. The F88/89A and F88/89A-W90A proteins exhibited reduced calcium dependent interaction with phenyl-Sepharose when compared with recS100A1, demonstrating that the carboxyl-terminal aromatic residues Phe 88, Phe 89, and Trp 90 comprise the PSBR of S100A1. Fluorescence studies showed that the Delta85 93 protein exhibited reduced calcium-dependent interaction with the dodecyl CapZ peptide, TRTK, while W90A bound TRTK with a Kd of 5.55 microM. These results demonstrate that the calcium-dependent target protein-binding site and the PSBR are indistinguishable. In contrast to the calcium-dependent target TRTK, activation of the calcium-independent target protein aldolase A by the point and deletion mutant S100A1s was indistinguishable from native S100A1. These results demonstrate that carboxyl-terminal residues are not required for S100A1 modulation of calcium-independent target protein aldolase A. Alltogether, these results indicate that S100A1 utilizes distinct mechanisms for interaction with calcium-independent and calcium-dependent target proteins. PMID- 9860859 TI - Modulation of quantum yield of primary radical pair formation in photosystem II by site-directed mutagenesis affecting radical cations and anions. AB - Pigment-protein interactions play a significant role in determining the properties of photosynthetic complexes. Site-directed mutants of Synechocystis PCC 6803 have been prepared which modify the redox potential of the primary radical pair anion and cation. In one set of mutants, the environment of P680, the primary electron donor of Photosystem II, has been modified by altering the residue at D1-His198. It has been proposed that this residue is an axial ligand to the magnesium cation. In the other set, the D1-Gln130 residue, which is thought to interact with the C9-keto group of the pheophytin electron acceptor, has been changed. The effect of these mutations is to alter the free energy of the primary radical pair state, which causes a change in the equilibrium between excited singlet states and radical pair states. We show that the free energy of the primary radical pair can be increased or decreased by modifications at either the D1-His198 or the D1-Gln130 sites. This is demonstrated by using three independent measures of quantum yield and equilibrium constant, which exhibit a quantitative correlation. These data also indicate the presence of a fast nonradiative decay pathway that competes with primary charge separation. These results emphasize the sensitivity of the primary processes of PS II to small changes in the free energy of the primary radical pair. PMID- 9860860 TI - Identification of the heme-modified peptides from cumene hydroperoxide inactivated cytochrome P450 3A4. AB - Cumene hydroperoxide-mediated (CuOOH-mediated) inactivation of cytochromes P450 (CYPs) results in destruction of their prosthetic heme to reactive fragments that irreversibly bind to the protein. We have attempted to characterize this process structurally, using purified, 14C-heme labeled, recombinant human liver P450 3A4 as the target of CuOOH-mediated inactivation, and a battery of protein characterization approaches [chemical (CNBr) and proteolytic (lysylendopeptidase C) digestion, HPLC-peptide mapping, microEdman sequencing, and mass spectrometric analyses]. The heme-peptide adducts isolated after CNBr/lysylendopeptidase-C digestion of the CuOOH-inactivated P450 3A4 pertain to two distinct P450 3A4 active site domains. One of the peptides isolated corresponds to the proximal helix L/Cys-region peptide 429-450 domain and the others to the K-region (peptide 359-386 domain). Although the precise residue(s) targeted remain to be identified, we have narrowed down the region of attack to within a 17 amino acid peptide (429-445) stretch of the 55-amino acid proximal helix L/Cys domain. Furthermore, although the exact structures of the heme-modifying fragments and the nature of the adduction remain to be established conclusively, the incremental masses of approximately 302 and 314 Da detected by electrospray mass spectrometric analyses of the heme-modified peptides are consistent with a dipyrrolic heme fragment comprised of either pyrrole ring A-D or B-C, a known soluble product of peroxidative heme degradation, as a modifying species. PMID- 9860861 TI - In vitro reconstitution of the core and peripheral light-harvesting complexes of Rhodospirillum molischianum from separately isolated components. AB - In most purple bacteria, the core light-harvesting complex (LH1) differs from the peripheral light-harvesting complex (LH2) in spectral properties and amino acid sequences. In Rhodospirillum (Rs. )molischianum, however, the LH2 closely resembles the LH1 of many species in amino acid sequence identity and in some spectral properties (e.g., circular dichroism and resonance Raman). Despite these similarities to LH1, the LH2 of Rs. molischianum displays an absorption spectrum similar to the LH2 complexes of other bacteria. Moreover, its crystal structure is very similar to the LH2 of Rhodopseudomonas (Rps.) acidophila. To better understand the basis of the biochemical and spectral differences between LH1 and LH2, we isolated the alpha and beta polypeptides of the LH2 complexes from an LH2 only strain of Rhodobacter (Rb.) sphaeroides as well as the alpha and beta polypeptides from both the LH1 and LH2 complexes from Rs. molischianum. We then examined their behavior in reconstitution assays with bacteriochlorophyll (Bchl). The Rb. sphaeroides LH2 alpha and beta polypeptides were inactive in reconstitution assays, whether alone, paired with each other, or paired in hybrid assays with the complementary LH1 polypeptides of Rs. rubrum, Rb. sphaeroides, Rb. capsulatus, or Rps. viridis. The LH1 beta polypeptide of Rs. molischianum behaved similarly to the LH1 beta polypeptides of Rs. rubrum, Rb. sphaeroides, Rb. capsulatus, and Rps. viridis, forming a subunit-type complex with or without an alpha polypeptide, and forming an LH1 complex when combined with a native LH1 alpha polypeptide. Interestingly, the LH2 beta polypeptide of Rs. molischianum, in the absence of other polypeptides, also formed a subunit-type complex as well as a further red-shifted complex whose spectrum resembled the 850 nm absorbance band of LH2. In the presence of the LH1 alpha polypeptide of Rs. rubrum or Rs. molischianum, it formed an LH1-type complex, but in the presence of the LH2 alpha polypeptide of Rs. molischianum it formed an LH2 complex. This is the first reported reconstitution of an LH2 complex using only isolated LH2 polypeptides and Bchl. It is also the first example of an LH2 beta polypeptide that can form an LH1 subunit-type complex and an LH1-type complex when paired with an LH1 alpha polypeptide. PMID- 9860862 TI - Generation of triplet and cation-radical bacteriochlorophyll a in carotenoidless LH1 and LH2 antenna complexes from Rhodobacter sphaeroides. AB - The LH1 antenna complex and a native form of the LH2 complex were isolated from the carotenoidless R26 and R26.1 mutants of Rhodobacter sphaeroides by the use of a new detergent, sucrose monocholate. One-color, pump-and-probe transient Raman spectroscopy of these complexes using 351 nm, approximately 50 ps pulses showed the generation of the triplet state of bacteriochlorophyll a (BChl a), whereas measurements using 355 nm, approximately 12 ns pulses showed the generation of BChl a cation radical. Subpicosecond to nanosecond time-resolved absorption spectroscopy using 388 nm, 200 fs pulses for excitation showed rapid (<1 ps) generation of the triplet state and fast decay (<10 ps) of the singlet state of BChl a. Microsecond absorption spectroscopy confirmed the generation of BChl a cation radical. EPR spectroscopy using 532 nm, approximately 5 ns pulses for excitation established the generation of BChl a cation radical. The EPR line width suggested that the unpaired electron is shared by two BChl a molecules. In LH1, the yield of BChl a cation radical per complex was estimated to be about 80% of that in the reaction center, and in LH2 about 50%. Thus, rapid generation of the triplet state, and its subsequent transformation into the cation-radical state of BChl a have been shown to be intrinsic properties of B870 and B850 BChl a assembly in the carotenoidless LH1 and LH2 antenna complexes. In the case of the carotenoid-containing LH2 complex, the triplet states of BChl a and carotenoid (spheroidene) were generated immediately after excitation, but the triplet-state BChl a was quenched efficiently by the carotenoid so that no BChl a cation radical was generated. Thus, the photoprotective function of the carotenoid in this antenna complex is shown. PMID- 9860863 TI - Mutagenesis studies of human red opsin: trp-281 is essential for proper folding and protein-retinal interactions. AB - Human red and green opsins contain a strikingly large number of tryptophan residues. These tryptophans are highly conserved among all red and green opsins. To investigate possible roles of these tryptophans in folding and structure, we have systematically replaced each tryptophan of human red opsin. When expressed in COS cells, wild-type red opsin undergoes N-linked glycosylation, forms a light sensitive pigment with absorption maximum at 560 nm upon reconstitution with 11 cis-retinal, and is transported to the plasma membrane. We used the extent of glycosylation, pigment generation, and intracellular localization of mutant red opsins as our criteria for assessing the effect of substitution. Replacement of eight tryptophans, Trp-59, Trp-90, Trp-149, Trp-152, Trp-183, Trp-191, Trp-195, and Trp-243, with Phe or Ala did not affect the wild-type phenotype significantly. However, replacement of Trp-5 and Trp-51 in the putative N terminal domain and Trp-142, Trp-177, Trp-179, and Trp-281 in the transmembrane domain with Phe had profound effects, indicating that these substitutions affected red opsin folding. Judged by the severity of the effects, we propose that Trp-5, Trp-51, Trp-177, and Trp-281 are important for red opsin folding. Although substitution of Trp-281 with Phe and Cys did not permit normal glycosylation and transport, substitution with Tyr and His permitted these processes but resulted in blue-shifted pigment. Thus, polar aromatics appear to substitute for Trp-281 to allow red opsin folding. The large spectral shift indicates that Trp-281 is essential for the proper interaction of the protein with 11-cis-retinal. PMID- 9860864 TI - Effects on substrate reduction of substitution of histidine-195 by glutamine in the alpha-subunit of the MoFe protein of Azotobacter vinelandii nitrogenase. AB - Studies of the substrate-reducing capabilities of an altered nitrogenase MoFe protein (alpha-195(Gln) instead of alpha-195(His)) from a mutant of Azotobacter vinelandii show, contrary to an earlier report [Kim, C.-H., Newton, W. E., and Dean, D. R. (1995) Biochemistry 34, 2798-2808], that the alpha-195(Gln) MoFe protein can reduce N2 to NH3 but at a rate that is <2% of that of the wild type. The extent of effective binding of N2 by this altered MoFe protein, as monitored by the inhibition of H2 evolution, is markedly increased as temperature is lowered but virtually eliminated at 45 degreesC. This inhibition of H2 evolution results in an increase in the ATP:2e- ratio, i.e., the number of molecules of MgATP hydrolyzed for each electron pair transferred to substrate, from ca. 5 (the wild-type level) at 45 degreesC to nearly 25 at 13 degreesC. Like wild-type nitrogenase, the N2 inhibition of H2 evolution reaches a maximum at an Fe protein:MoFe protein molar ratio of ca. 2.5, suggesting that a highly reduced enzyme may not be necessary for N2 binding. N2 binding to the alpha-195(Gln) MoFe protein retains a hallmark of the wild type by producing HD under a mixed N2/D2 atmosphere. The rate of HD production and the fraction of total electron flow allocated to HD are similar to those for wild-type nitrogenase under the same conditions. However, the electrons forming HD do not come from those normally producing NH3 (as occurs in the wild type) but are equivalent to those whose evolution as H2 had been inhibited by N2. N2 also inhibits C2H2 reduction catalyzed by the alpha-195(Gln) nitrogenase. This inhibition is relieved by added H2, resulting in a lowering of the elevated ATP:2e- ratio to that found under Ar. With solutions of NaCN, which contain both the substrate, HCN, and the inhibitor, CN-, reduction of HCN is not impaired with the alpha-195(Gln) nitrogenase, but the inhibition by CN- of total electron flow to substrate, which is observed with the wild-type MoFe protein, is completely absent. Unlike that of the catalyzed reduction of H+, HCN, or C2H2, the extent of azide reduction to either N2 or N2H4 is markedly decreased (to 5-7% of that of the wild type) with the alpha-195(Gln) nitrogenase. Azide, like N2, inhibits H2 evolution and increases the ATP:2e- ratio. Both effects are freely reversible and abolished by CO. Added D2 does not relieve either effect, implying that N2 produced from N3- is not the inhibitory species. The correlation between the extremely low rates of reduction for both N2 and azide by the alpha-195(Gln) nitrogenase and their common ability to inhibit H2 evolution suggests that alpha-histidine-195 may be an important proton conductor to the FeMo cofactor center and specifically required for reduction of these two substrates. PMID- 9860865 TI - Inhibition of elastase by N-sulfonylaryl beta-lactams: anatomy of a stable acyl enzyme complex. AB - beta-Lactam inhibitors of transpeptidase enzymes involved in cell wall biosynthesis remain among the most important therapeutic agents in clinical use. beta-Lactams have more recently been developed as inhibitors of serine proteases including elastase. All therapeutically useful beta-lactam inhibitors operate via mechanisms resulting in the formation of hydrolytically stable acyl-enzyme complexes. Presently, it is difficult to predict which beta-lactams will form stable acyl-enzyme complexes with serine enzymes. Further, the factors that result in the seemingly special nature of beta-lactams versus other acylating agents are unclear-if indeed they exist. Here we present the 1.6 A resolution crystal structure of a stable acyl-enzyme complex formed between porcine pancreatic elastase and a representative monocyclic beta-lactam, which forms a simple acyl-enzyme. The structure shows that the ester carbonyl is not located within the oxyanion hole and the "hydrolytic" water is displaced. Combined with additional kinetic and mass spectrometric data, the structure allows the rationalization of the low degree of hydrolytic lability observed for the beta lactam-derived acyl-enzyme complex. PMID- 9860866 TI - Substrates and inhibitors of human T-cell leukemia virus type I protease. AB - HTLV-I is an oncogenic retrovirus that is associated with adult T-cell leukemia. HTLV-I protease and HTLV-I protease fused to a deca-histidine containing leader peptide (His-protease) have been cloned, expressed, and purified. The refolded proteases were active and exhibited nearly identical enzymatic activities. To begin to characterize the specificity of HTLV-I, we measured protease cleavage of peptide substrates and inhibition by protease inhibitors. HTLV-I protease cleavage of a peptide representing the HTLV-I retroviral processing site P19/24 (APQVLPVMHPHG) yielded Km and kcat values of 470 microM and 0.184 s-1 while cleavage of a peptide representing the processing site P24/15 (KTKVLVVQPK) yielded Km and kcat values of 310 microM and 0.0060 s-1. When the P1' proline of P19/24 was replaced with p-nitro-phenylalanine (Nph), the ability of HTLV-I protease to cleave the substrate (APQVLNphVMHPL) was improved. Inhibition of HTLV I protease and His-protease by a series of protease inhibitors was also tested. It was found that the Ki values for inhibition of HTLV-I protease and His protease by a series of pepsin inhibitors ranged from 7 nM to 10 microM, while the Ki values of a series of HIV-1 protease inhibitors ranged from 6 nM to 127 microM. In comparison, the Ki values for inhibition of pepsin by the pepsin inhibitors ranged from 0.72 to 19.2 nM, and the Ki values for inhibition of HIV-1 protease by the HIV protease inhibitors ranged from 0.24 nM to 1.0 microM. The data suggested that the substrate binding site of HTLV-I protease is different from the substrate binding sites of pepsin and HIV-1 protease, and that currently employed HIV-1 protease inhibitors would not be effective for the treatment of HTLV-I infections. PMID- 9860867 TI - Topological and functional relationship of subunits F1-gamma and F0I-PVP(b) in the mitochondrial H+-ATP synthase. AB - Diamide treatment of the F0F1-ATP synthase in "inside out" submitochondrial particles (ESMP) in the absence of a respiratory Delta mu H+ as well as of isolated Fo reconstituted with F1 or F1-gamma subunit results in direct disulfide cross-linking between cysteine 197 in the carboxy-terminal region of the F0I PVP(b) subunit and cysteine 91 at the carboxyl end of a small alpha-helix of subunit F1-gamma, both located in the stalk. The F0I-PVP(b) and F1-gamma cross linking cause dramatic enhancement of oligomycin-sensitive decay of Delta mu H+. In ESMP and MgATP particles the cross-linking is accompanied by decoupling of respiratory ATP synthesis. These effects are consistent with the view that F0I PVP(b) and F1-gamma are components of the stator and rotor of the proposed rotary motor, respectively. The fact that the carboxy-terminal region of F0I-PVP(b) and the short alpha-helix of F1-gamma can form a direct disulfide bridge shows that these two protein domains are, at least in the resting state of the enzyme, in direct contact. In isolated F0, diamide also induces cross-linking of OSCP with another subunit of F0, but this has no significant effect on proton conduction. When ESMP are treated with diamide in the presence of Delta mu H+ generated by respiration, neither cross-linking between F0I-PVP(b) and F1-gamma subunits nor the associated effects on proton conduction and ATP synthesis is observed. Cross linking is restored in respiring ESMP by Delta mu H+ collapsing agents as well as by DCCD or oligomycin. These observations indicate that the torque generated by Delta mu H+ decay through Fo induces a relative motion and/or a separation of the F0I-PVP(b) subunit and F1-gamma which places the single cysteine residues, present in each of the two subunits, at a distance at which they cannot be engaged in disulfide bridging. PMID- 9860868 TI - Spectroscopic evidence for nanosecond protein relaxation after photodissociation of myoglobin-CO. AB - Nanosecond time-resolved absorption and magnetic optical rotatory dispersion (MORD) measurements of photolyzed myoglobin-CO visible bands (500-650 nm) are presented. These measurements reveal a 400 ns process, spectrally distinct from ligand recombination, that accounts for 7% of the observed spectral evolution in the visible absorption bands and 4% in the MORD. The time-resolved MORD, more sensitive to heme coordination geometry than absorption, suggests that this process is most likely associated with protein relaxation on the distal side of the heme pocket, perhaps accompanying rehydration of the deoxymyoglobin photoproduct or accommodation of protein side chains to ligand escape. PMID- 9860869 TI - The unusually slow unfolding rate causes the high stability of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus: equilibrium and kinetic studies of guanidine hydrochloride-induced unfolding and refolding. AB - To elucidate the energetic features of the anomalously high-level stabilization of a hyperthermophile pyrrolidone carboxyl peptidase (PfPCP) from a hyperthermophilic archaeon, Pyrococcus furiosus, equilibrium and kinetic studies of the guanidine hydrochloride (GuHCl)-induced unfolding and refolding were carried out with CD measurements at 220 nm in comparison with those from the mesophile homologue (BaPCP) from Bacillus amyloliquefaciens. The mutant protein of PfPCP substituted with Ser at both Cys142 and Cys188 (PfC142/188S) was used. The GuHCl unfolding for PfC142/188S and BaPCP was reversible. It was difficult to obtain the equilibrated unfolding curve of the hyperthermophile proteins at temperatures below 50 degreesC and pH 7, because of the remarkably slow rate of the unfolding. The unfolding for PfC142/188S attained equilibrium after 7 days at 60 degreesC, resulting in the coincidence between the unfolding and refolding curves. The Gibbs energy change of unfolding, DeltaGH2O (56.6 kJ/mol), for PfC142/188S at 60 degreesC and pH 7 was dramatically higher than that (7.6 kJ/mol) for BaPCP at 40 degreesC and pH 7. The unfolding and refolding kinetics for PfC142/188S and BaPCP at both 25 and 60 degreesC at pH 7 were approximated as a single exponential. The rate constant in water (kuH2O) of the unfolding reaction for PfC142/188S (1.6 x 10(-)15 s-1) at 25 degreesC and pH 7 was drastically reduced by 7 orders of magnitude compared to that (1.5 x 10(-)8 s-1) for BaPCP, whereas the refolding rates (krH2O) in water for PfC142/188S (9.3 x 10(-)2 s-1) and BaPCP (3.6 x 10(-)1 s-1) at 25 degreesC and pH 7 were similar. These results indicate that the greater stability of the hyperthermophile PCP was characterized by the drastically slow unfolding rate. PMID- 9860870 TI - Sensitivity of flavin fluorescence dynamics in neuronal nitric oxide synthase to cofactor-induced conformational changes and dimerization. AB - The fluorescence intensity of the two flavin prosthetic groups, FMN and FAD, in neuronal nitric oxide synthase (nNOS) was found to decay highly nonexponentially, being best described by four fluorescence lifetimes. This excited state heterogeneity is the result of multiple flavin quenching sites which are due to several flavin microenvironments created mainly by stacking with aromatic amino acids. Investigating nNOS in the absence of one or more of Ca2+/calmodulin, tetrahydrobiopterin, and heme revealed an influence of these cofactors on the microenvironments of the flavin prosthetic groups. Similar effects on the flavin rotational dynamics were found by analyzing the fluorescence anisotropy decay of the holo and of the different apo forms of nNOS. Since the tetrahydrobiopterin and the heme are located in the N-terminal oxygenase domain of nNOS, their effect on the flavins in the C-terminal reductase domain is explained by a folding back of the reductase domain onto the oxygenase domain. Thereby a domain-domain interface is created containing the FAD, FMN, heme, and tetrahydrobiopterin prosthetic groups which allows for efficient electron transfer during catalysis. The heme group, which is known to be essential for homodimerization of nNOS, was also found to be essential for the formation of the domain-domain interface. PMID- 9860871 TI - Lipid properties and the orientation of aromatic residues in OmpF, influenza M2, and alamethicin systems: molecular dynamics simulations. AB - Molecular dynamics simulations allow a direct study of the structure and dynamics of membrane proteins and lipids. We describe the behavior of aromatic residues and lipid properties in POPE and POPC bilayer models with the Escherichia coli OmpF trimer, single alamethicin and Influenza M2 helices, 4-helix M2 bundles, and two alamethicin 6-helix channel models. The total simulation time is over 24 ns, of systems containing solvent, protein, and between 104 and 318 lipids. Various types of adjustment between lipids and proteins occur, depending on the size of the protein and the degree of hydrophobic mismatch between lipid and protein. Single helices cause little measurable effect on nearby lipids whereas the 4 helix bundles, 6-helix channel models, and OmpF cause a significant lowering of order parameters in nearby lipid chains, an increased difference between odd and even chain dihedrals in the magnitude of the trans dihedral fractions and dihedral transition rates, and in most cases a decreased gauche population and a decrease in bilayer thickness. An increased tilt of the lipid chains near the proteins can account for most of the observed decrease in order parameters. The orientation of tryptophans and tyrosines on the outside of the proteins is determined by packing at the protein exterior and non-specific hydrogen bonding with lipids and solvent. The tyrosines in the broad bands that delimit the hydrophobic exterior of OmpF show little change in orientation over one nanosecond. Their rings are oriented predominantly perpendicular to the bilayer plane, with the hydroxyl group pointing toward the lipid-water interface. Phenylalanines in OmpF, alamethicin, and Influenza M2 are more mobile and assume a variety of orientations. PMID- 9860872 TI - Sphingomyelinase induces lipid microdomain formation in a fluid phosphatidylcholine/sphingomyelin membrane. AB - The behaviors of two chemically well-defined sphingolipids, N-palmitoyl sphingomyelin (C16:0-SM) and the corresponding ceramide (C16:0-Cer), in a 1 palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) matrix were compared. Minor attenuation of lateral diffusion upon increasing the mole fraction of C16:0 SM (XSM, up to 0.25) was indicated by the slight decrement in the excimer/monomer intensity ratio (Ie/Im) for a trace amount (mole fraction X = 0.01) of a pyrene labeled ceramide analogue (N-[(pyren)-1-yl]decanoyl-sphingosine, PDCer) in keeping with the miscibility of C16:0-SM in POPC. Increasing membrane order was revealed by the augmented polarization P for diphenylhexatriene (DPH). In contrast, when C16:0-Cer was substituted for C16:0-SM an approximately 1.6-fold increase in Ie/Im for PDCer was evident upon increasing Xcer, with parallel increment in DPH polarization. In agreement with our recent data on natural ceramides in dimyristoylphosphatidylcholine (DMPC) bilayers [Holopainen et al. (1997) Chem. Phys. Lipids 88, 1-13], we conclude that C16:0-Cer becomes enriched into microdomains in the fluid POPC membrane. Interestingly, enhanced formation of microdomains by ceramide was observed when the total sphingolipid content in tertiary alloys with POPC was maintained constant (Xcer + XSM = 0.25) and the SM/Cer stoichiometry was varied. Finally, when ceramide was generated enzymatically in POPC/C16:0-SM (3:1, molar fraction) LUVs by sphingomyelinase (SMase, Bacillus cereus), maximally approximately 85% of hydrolysis of sphingomyelin was measured within <3 min at 30 degreesC. The formation of ceramide was accompanied by a closely parallel increase in DPH polarization. There was also an increase in Ie/Im for PDCer; however, these changes in Ie/Im were significantly slower, requiring approximately 105 min to reach a steady state. These data show that the rapid enzymatic formation of ceramide under these conditions is followed by much slower reorganization process, resulting in the formation of microdomains enriched in this lipid. PMID- 9860873 TI - Suppression of microtubule dynamics by binding of cemadotin to tubulin: possible mechanism for its antitumor action. AB - Cemadotin (LU103793) (NSC D-669356) is a water-soluble synthetic analogue of dolastatin 15 that inhibits cell proliferation in vitro and the growth of human tumor xenografts. Cemadotin is in phase II clinical trials as a promising cancer chemotherapeutic agent. The drug blocks cells at mitosis. Its primary mode of action has been unclear but is believed to involve an action on microtubules. We have found that cemadotin binds to tubulin and strongly suppresses microtubule dynamics. Scatchard analysis of cemadotin binding to tubulin indicated that there are two affinity classes of cemadotin-binding sites with Kd values of 19.4 microM and 136 microM. Cemadotin did not inhibit the binding of vinblastine to tubulin, and, conversely, vinblastine did not inhibit the binding of cemadotin to tubulin. By quantitative video microscopy of individual microtubules, we found that cemadotin strongly suppressed dynamic instability of microtubules assembled to steady state using bovine brain tubulin devoid of microtubule-associated proteins. It reduced the rate and extent of growing and shortening, increased the rescue frequency, and increased the percentage of time the microtubules spent in an attenuated or paused state, neither growing nor shortening detectably. At the lowest effective cemadotin concentrations, dynamics were suppressed in the absence of significant microtubule depolymerization. The results suggest that cemadotin exerts its antitumor activity by suppressing spindle microtubule dynamics through a distinct molecular mechanism by binding at a novel site in tubulin. PMID- 9860874 TI - Evidence for kinetic intermediate states during the refolding of GdnHCl-denatured MM-creatine kinase. Characterization of a trapped monomeric species. AB - The kinetics of refolding of guanidinium chloride-denatured rabbit MM-creatine kinase was investigated. Recovery of enzymatic activity is biphasic, depending on the temperature but not on the protein or DTT concentration. Only 45% of the original, active dimeric form is recovered even after several hours of refolding. The reactivation yield is limited by the accumulation of a highly stable but nonproductive monomeric species. The ratio of "correct" to "incorrect" forms depends on the duration of exposure to the denaturant, which may be consistent with the existence of a heterogeneous population of unfolded states with regard to proline isomerization. The first fast reaction observed during renaturation results in the appearance of collapsed monomeric states, displaying features of a pre-molten globule state. These burst species are rapidly transformed into more structured monomers resembling a molten globule state possessing a partially folded C-terminal domain. A proportion of these latter transient intermediates (45%) associates into an active dimer, while the remainder (55%) is trapped by reshuffling in a monomeric dead-end product. Our results strongly indicate that (i) the dimeric state is a prerequisite for the expression of catalytic activity, (ii) the kinetic intermediates of refolding are very similar to those observed during equilibrium unfolding, and (iii) refolding of creatine kinase in these conditions is limited by the accumulation of inactive misfolded nondimerizable monomer. PMID- 9860875 TI - Conversion of a catalytic into a structural disulfide bond by circular permutation. AB - The thiol-disulfide oxidoreductase DsbA from Escherichia coli is the strongest oxidant of the enzyme family and required for disulfide bond formation in the bacterial periplasm. The catalytic domain of this 189-residue protein has a thioredoxin-like fold and contains a catalytic disulfide bridge that is located within the sequence Cys30-Pro31-His32-Cys33 at the N-terminus of an alpha-helix. The Cys30-Cys33 disulfide bond destabilizes DsbA by about 16 kJ/mol at pH 7.0, which appears to be caused by the extremely low pKa value of approximately 3.4 of the nucleophilic Cys30 thiol. Here we report the characterization of a circularly permuted variant of DsbA, termed H32-P31, in which the natural termini are connected by a Gly3-Thr-Gly linker and the new termini are located between the active-site cysteines (first residue His32, last residue Pro31). The disulfide bond in the variant thus connects the second with the penultimate residue. H32 P31 adopts a wild-type-like structure and folds reversibly and cooperatively in both redox forms. However, the permuted variant is catalytically inactive as dithiol oxidase in vivo and in vitro. Both cysteine thiols have pKa values > 8; the variant is 500-fold more reducing than the wild type and more stable in its oxidized form. Thus, the Cys30-Cys33 disulfide in the variant H32-P31 has adopted properties of a structural disulfide bond. PMID- 9860876 TI - Porcine recombinant dihydropyrimidine dehydrogenase: comparison of the spectroscopic and catalytic properties of the wild-type and C671A mutant enzymes. AB - Dihydropyrimidine dehydrogenase catalyzes, in the rate-limiting step of the pyrimidine degradation pathway, the NADPH-dependent reduction of uracil and thymine to dihydrouracil and dihydrothymine, respectively. The porcine enzyme is a homodimeric iron-sulfur flavoprotein (2 x 111 kDa). C671, the residue postulated to be in the uracil binding site and to act as the catalytically essential acidic residue of the enzyme oxidative half-reaction, was replaced by an alanyl residue. The mutant enzyme was overproduced in Escherichia coli DH5alpha cells, purified to homogeneity, and characterized in comparison with the wild-type species. An extinction coefficient of 74 mM-1 cm-1 was determined at 450 nm for the wild-type and mutant enzymes. Chemical analyses of the flavin, iron, and acid-labile sulfur content of the enzyme subunits revealed similar stoichiometries for wild-type and C671A dihydropyrimidine dehydrogenases. One FAD and one FMN per enzyme subunit were found. Approximately 16 iron atoms and 16 acid-labile sulfur atoms were found per wild-type and mutant enzyme subunit. The C671A dihydropyrimidine dehydrogenase mutant exhibited approximately 1% of the activity of the wild-type enzyme, thus preventing its steady-state kinetic analysis. Therefore, the ability of the C671A mutant and, for comparison, of the wild-type enzyme species to interact with reaction substrates, products, or their analogues were studied by absorption spectroscopy. Both enzyme forms did not react with sulfite. The wild-type and mutant enzymes were very similar to each other with respect to the spectral changes induced by binding of the reaction product NADP+ or of its nonreducible analogue 3-aminopyridine dinucleotide phosphate. Uracil also induced qualitatively and quantitatively similar absorbance changes in the visible region of the absorbance spectrum of the two enzyme forms. However, the calculated Kd of the enzyme-uracil complex was significantly higher for the C671A mutant (9.1 +/- 0.7 microM) than for the wild type dihydropyrimidine dehydrogenase (0.7 +/- 0.09 microM). In line with these observations, the two enzyme forms behaved in a similar way when titrated anaerobically with a NADPH solution. Addition of an up to 10-fold excess of NADPH to both dihydropyrimidine dehydrogenase forms led to absorbance changes consistent with reduction of approximately 0.5 flavin per subunit, with no indication of reduction of the enzyme iron-sulfur clusters. Absorbance changes consistent with reduction of both enzyme flavins were obtained by removing NADP+ with a NADPH-regenerating system. On the contrary, the two enzyme species differed significantly with respect to their reactivity with dihydrouracil. Addition of dihydrouracil to the wild-type enzyme species, under anaerobic conditions, led to absorbance changes that could be interpreted to result from both partial flavin reduction and the formation of a complex between the enzyme and (dihydro)uracil. In contrast, only spectral changes consistent with formation of a complex between the oxidized enzyme and dihydrouracil were observed when a C671A mutant enzyme solution was titrated with this compound. Furthermore, enzyme monitored turnover experiments were carried out anaerobically in the presence of a limiting amount of NADPH and excess uracil with the two enzyme forms in a stopped-flow apparatus. These experiments directly demonstrated that the substitution of an alanyl residue for C671 in dihydropyrimidine dehydrogenase specifically prevents enzyme-catalyzed reduction of uracil. Finally, sequence analysis of dihydropyrimidine dehydrogenase revealed that it exhibits a modular structure; the N-terminal region, similar to the beta subunit of bacterial glutamate synthases, is proposed to be responsible for NADPH binding and oxidation with reduction of the FAD cofactor of dihydropyrimidine dehydrogenase. The central region, similar to the FMN subunit of dihydroorotate dehydrogenases, is likely to harbor the site o PMID- 9860877 TI - Identification of two electron-transfer sites in ascorbate peroxidase using chemical modification, enzyme kinetics, and crystallography. AB - Chemical and mutagenic modification combined with X-ray crystallography has been used to probe the ascorbate binding site in ascorbate peroxidase (APX). Chemical modification of the single Cys residue in APX with Ellman's reagent (DTNB) blocks the ability of APX to oxidize ascorbate but not other small aromatic phenolic substrates. DTNB-modified APX (APX-TNB) exhibits only 1.3% wild-type activity when ascorbate is used as the substrate but full activity when aromatic substrates, guaiacol or pyrogallol, are used. Stopped-flow studies show that APX TNB reacts normally with peroxide to give compound I but that the rates of reduction of both compounds I and II by ascorbate are dramatically slowed. Conversion of Cys32 to Ser leads to approximately 70% drop in ascorbate peroxidase activity with no effect on guaiacol peroxidase activity. These results indicate that uncharged aromatic substrates and the anionic ascorbate molecule interact with different sites on APX. The 2.0 A X-ray crystal structure of APX TNB shows clear electron density for the TNB group covalently attached to Cys32 in all four molecules of the asymmetric unit, indicating complete and specific modification. It appears that the ascorbate site is blocked by DTNB modification which is well removed from the exposed delta-heme edge where aromatic substrates are thought to bind. This is the first experimental evidence indicating that ascorbate oxidation does not occur at the exposed heme edge but at an alternate binding site in the vicinity of Cys32 near Arg172 and the heme propionates. PMID- 9860878 TI - Identification of individual nucleotides in the bacterial ribonuclease P ribozyme adjacent to the pre-tRNA cleavage site by short-range photo-cross-linking. AB - The bacterial RNase P ribozyme is a site-specific endonuclease that catalyzes the removal of pre-tRNA leader sequences to form the 5' end of mature tRNA. While several specific interactions between enzyme and substrate that direct this process have been determined, nucleotides on the ribozyme that interact directly with functional groups at the cleavage site are not well-defined. To identify individual nucleotides in the ribozyme that are in close proximity to the pre tRNA cleavage site, we introduced the short-range photoaffinity cross-linking reagent 6-thioguanosine (s6G) at position +1 of tRNA and position -1 in a tRNA bearing a one-nucleotide leader sequence [tRNA(G-1)] and examined cross-linking in representatives of the two structural classes of bacterial RNase P RNA (from Escherichia coli and Bacillus subtilis). These photoagent-modified tRNAs bind with similar high affinity to both ribozymes, and the substrate bearing a single s6G upstream of the cleavage (-1) site is cleaved accurately. Interestingly, s6G at position +1 of tRNA cross-links with high efficiency to homologous positions in J5/15 in both E. coli and B. subtilis RNase P RNAs, while s6G at position -1 of tRNA(G-1) cross-links to homologous nucleotides in J18/2. Both cross-links are detected over a range of ribozyme and substrate concentrations, and importantly, ribozymes cross-linked to position -1 of tRNA(G-1) accurately cleave the covalently attached substrate. These data indicate that the conserved guanosine at the 5' end of tRNA is adjacent to A248 (E. coli) of J5/15, while the base upstream of the substrate phosphate is adjacent to G332 (E. coli) of J18/2 and, along with available biochemical data, suggest that these nucleotides play a direct role in binding the substrate at the cleavage site. PMID- 9860879 TI - Folding of the four-way RNA junction of the hairpin ribozyme. AB - The hairpin ribozyme consists of two loop-carrying duplexes (called A and B) that are adjacent arms of a four-way junction in its natural context in the viral RNA. We have shown previously that the activity of the ribozyme is strongly influenced by the structure adopted by the junction. In this study, we have used fluorescence resonance energy transfer to analyze the conformation and folding of the isolated four-way junction. Like other four-way RNA junctions, in the absence of added metal ions this junction adopts a square configuration of coaxially stacked arms, based on A on D and B on C stacking. Upon addition of magnesium ions, the junction undergoes an ion-induced transition to an antiparallel conformation. The data are consistent with folding induced by the binding of a single ion, with an apparent association constant in the range of 2000 M-1. Other divalent metal ions (calcium or manganese) can also induce this change in structure; however, sodium ions are unable to substitute for these ions, and are slightly inhibitory with respect to the transition. The loop-free hairpin junction adopts the same stacking conformer as the full ribozyme, but forms a more symmetrical X-shaped structure. In addition, the apparent stoichiometry of structural ion binding is lower for the isolated junction, and the affinity is considerably lower. PMID- 9860880 TI - Human health and chemical mixtures: an overview. AB - Unlike laboratory animals, people are rarely exposed to a single hazardous chemical. However, most of the information documenting adverse human health effects from environmental and occupational contaminants has come from studies focused on exposure to single chemicals, and there is little information available on how two or more contaminants affect humans. Most information on the effects of mixtures comes from animal systems and limited investigations of isolated human cells in culture, even though the study of mixtures in such systems has also been neglected. Two or more compounds may show additive, antagonistic, or synergistic interactions or may act on totally different systems and thus not interact. Furthermore, even a single chemical may have multiple effects and affect more than one organ system. Effects may vary with age, and metabolites may have totally different actions from the parent compound. This paper will review the variety of health effects in humans that may result from environmental contaminants and discuss how such contaminants may interact with each other. We will also present examples on how different contaminants interact from toxicologic studies of polychlorinated biphenyls performed as part of our Albany, New York, Superfund Basic Research Program project. PMID- 9860881 TI - Public health challenges posed by chemical mixtures. AB - Approximately 40 million people live within a 4-mile radius of waste sites that the Agency for Toxic Substances and Disease Registry (ATSDR) has assessed to date. Human populations living in the vicinity of such sites are often subjected to complex chemical exposures that may contribute to the total body burden of oxogenous chemicals. Apart from the contaminants found at waste sites, exposure may also include environmental, occupational, and personal agents. Concurrent exposure to chemicals such as welding fumes, indoor air pollutants, tobacco smoke, alcohol, and prescription and nonprescription drugs makes the health assessment of exposure to waste site chemicals a more complex task. Voluntary exposures such as these frequently entail exposures to relatively high chemical concentrations and can usually be well defined and quantified. Conversely, involuntary exposures from waste sites may be at low concentrations and hence difficult to characterize and quantify. Of the approximately 1450 waste sites evaluated by the ATSDR, 530 (37%) had either completed or potentially completed exposure pathways. Results of public health assessments conducted at 167 sites during 1993 to 1995 show that about 1.5 million people have been exposed to site specific contaminants. At 10% or more of the sites that had either completed or potentially completed exposure pathways, 56 substances were identified. Of these, 19 are either known or anticipated human carcinogens, and 9 are associated with reproductive or endocrine-disrupting effects. In this paper we present important concerns regarding hazardous waste sites including the impact on human health, ecology, and quality of life. To address such human-health related issues, the ATSDR has established a mixtures program that consists of three components: trend analysis to identify combinations of chemicals of concern, experimental studies to identify data that would be useful in the development and implementation of predictive decision support methodologies, and development of assessment methodologies and guidance to provide health assessors with the tools to incorporate the evaluation of multiple-chemical exposure into site assessments. PMID- 9860882 TI - Toxicology of chemical mixtures: international perspective. AB - This paper reviews major activities outside the United States on human health issues related to chemical mixtures. In Europe an international study group on combination effects has been formed and has started by defining synergism and antagonism. Successful research programs in Europe include the development and application of statistically designed experiments combined with multivariate data analysis and modeling in vitro and in vivo studies on a wide variety of chemicals such as petroleum hydrocarbons, aldehydes, food contaminants, industrial solvents, and mycotoxins. Other major activities focus on the development of safety evaluation strategies for mixtures such as the use of toxic equivalence factors or alternatives such as the question-and-answer approach, fractionation followed by recombination of the mixture in combination with a mixture design, and quantitative structure-activity relationship analysis combined with lumping analysis and physiologically based pharmacokinetic/pharmacodynamic modeling for studying complex mixtures. A scheme for hazard identification and risk assessment of complex mixtures and a consistent way to generate total volatile organic compound values for indoor air have also been developed. Examples of other activities are carcinogenicity studies on complex mixtures (petroleum middle distillates, foundry fumes, pesticides, heterocyclic amines, diesel exhaust, solid particles), neurotoxicity studies of mixtures of solvents alone or in combination with exposure to physical factors, and toxicity studies of outdoor air pollutants, focusing on particulates. Outside the United States, toxicologists and regulators clearly have a growing interest in the toxicology and risk assessment of chemical mixtures. PMID- 9860883 TI - Maximum workplace concentration values and carcinogenicity classification for mixtures. AB - In Germany, the Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area (MAK Commission) generally sets maximum workplace concentration values (i.e., a proposed occupational exposure level [OEL]) for single substances, not for mixtures. For mixtures containing substances with a genotoxic and carcinogenic potential, the commission considered it scientifically inappropriate to establish a safe threshold. This approach is currently under discussion. Carcinogenic mixtures are categorized according to either the carcinogenicity of the mixture or the classification of the carcinogenic substances included. In regulating exposure to mixtures, an approach similar to that used by the American Conference of Governmental Hygienists is proposed: For components with the same target organ and mode of action or interfering metabolism, synergistic effects must be expected and the respective OELs must be lowered. However, if there is proof that the components act independently, the OELs of the individual compounds are not considered to be modified. In the view of the commission, calculating OELs for solvent mixtures according to their liquid phase composition is not justified, and the setting of scientifically based OELs for complex mixtures is not possible. PMID- 9860884 TI - Current approaches toward chemical mixture studies at the National Institute of Environmental Health Sciences and the U.S. National Toxicology Program. AB - The National Institute of Environmental Health Sciences (NIEHS) has several new initiatives involving chemical mixtures and has recognized the need to develop new experimental approaches to enhance our efforts in this area. Responding to recent increases in nominations of complex occupational exposures for toxicologic assessment by the U.S. National Toxicology Program, the NIEHS and the National Institute for Occupational Safety and Health have begun a program to characterize exposures through field studies, identify biomarkers of exposure in workers, and recreate relevant mixed exposures in a laboratory setting. A second initiative with the National Center for Environmental Health/Centers for Disease Control and Prevention will examine blood samples from the U.S. National Health and Nutrition Examination Survey population surveys for selected endocrine-disrupting agents and for common patterns of persistent xenobiotics, providing critical information for the design of animal studies to assess risks of relevant chemical mixtures to humans. New toxicology testing methods (lower cost, faster) will enhance our ability to study chemical mixtures (e.g., dioxin and dioxinlike chemicals, combination AIDS therapies). Ongoing method development efforts involve in vitro functional toxicology assays, screens for estrogenic activity, and carcinogenesis studies in transgenic mice. A major scientific initiative with mixtures involves studies of individual and mixtures of dioxin and dioxinlike chemicals to determine if toxic equivalence factors predict carcinogenic potency in traditional and transgenic bioassays. Complementing these studies is an increased emphasis on physiologically based pharmacokinetic modeling, an activity central to the proper interpretation of chemical mixture studies. PMID- 9860885 TI - Environmental complex mixture toxicity assessment. AB - Trichloroethylene (TCE) was found as a contaminant in the well supplying water to an aquatic testing laboratory. The groundwater was routinely screened by a commercial laboratory for volatile and semivolatile compounds, metals, herbicides, pesticides, and polychlorinated biphenyls using U.S. Environmental Protection Agency methods. Although TCE was the only reportable peak on the gas chromatograph, with average concentrations of 0.200 mg/l, other small peaks were also present, indicating the possibility that the contamination was not limited to TCE alone. A chronic 6-month carcinogenicity assay was conducted on-site in a biomonitoring trailer, using the Japanese medaka fish (Oryzias latipes) in an initiation-promotion protocol, with diethylnitrosamine (DEN) as the initiator and the TCE-contaminated groundwater as a promoter. Study results indicated no evidence of carcinogenic potential of the groundwater without initiation. There was, however, a tumor-promotional effect of the groundwater after DEN initiation. A follow-up laboratory study was conducted using reagent grade TCE added to carbon-filtered groundwater to simulate TCE concentrations comparable to those found in the contaminated groundwater. Study results indicated no promotional effects of TCE. These studies emphasize the necessity for on-site bioassays to assess potential environmental hazards. In this instance, chemical analysis of the groundwater identified TCE as the only reportable contaminant, but other compounds present below reportable limits were noted and may have had a synergistic effect on tumor promotion observed with the groundwater exposure. Laboratory toxicity testing of single compounds can produce toxicity data specific to that compound for that species but cannot take into account the possible toxic effects of mixtures of compounds. PMID- 9860887 TI - Interactive toxicity and stress protein expression by vinylidene chloride and monochloroacetate in precision-cut rat liver slices. AB - Vinylidene chloride (VDC) is a groundwater and drinking water contaminant. Monochloroacetic acid (MCA) is a chlorination by-product of drinking water. Because environmental or occupational exposure to chemicals takes place at low concentrations, a sensitive in vitro system of liver slices was used to examine the interactive toxicity of MCA and VDC. Liver slices from Sprague-Dawley rats were exposed to 100 microM MCA for 1 hr before exposure to 20 or 48 microM VDC and incubated for 1 to 8 hr. MCA + 48 microM VDC resulted in a significant leakage of K+ by 4 hr, while MCA+ 20 microM VDC did not. At 4 hr, MCA + 48 microM VDC resulted in centrilobular necrosis. MCA caused a significant depletion of slice glutathione (GSH) at 1 hr, which was maintained up to 3 hr. As reactive VDC metabolites are detoxified by conjugation with GSH, the increase in VDC toxicity by MCA is possibly due to GSH-depleting effects of MCA. Heat shock protein (HSP) 72 was increased 2.5-fold by MCA + 20 microM VDC as early as 2 hr, although K+ leakage was not increased. MCA + 48 microM VDC resulted in a 3-fold increase in HSP 72 by 2 hr, while there were modest increases in HSPs 60 and 32. Therefore, HSP 72 is an early sensitive indicator of interactive toxicity of nontoxic concentrations of MCA and VDC. This is the first time that micromolar concentrations of these drinking water contaminants were observed to affect cellular homeostasis in the liver. PMID- 9860886 TI - Role of tissue repair in toxicologic interactions among hepatotoxic organics. AB - It is widely recognized that exposure to combinations or mixtures of chemicals may result in highly exaggerated toxicity even though individual chemicals might not be toxic at low doses. Chemical mixtures may also cause additive or less than additive toxicity. From the perspective of public health, highly exaggerated toxicity is of significant concern. Assessment of risk from exposure to chemical mixtures requires knowledge of the underlying mechanisms. Previous studies from this laboratory have shown that nontoxic doses of chlordecone (10 ppm, 15 days) and carbon tetrachloride (CCl4) (100 microliters/kg) interact at the biologic interface, resulting in potentiated liver injury and 67-fold amplification of CCl4 lethality. In contrast, although interaction between phenobarbital and CCl4 leads to even higher injury, animal survival is unaffected because of highly stimulated compensatory tissue repair. A wide variety of additional experimental evidence confirms the central role of stimulated tissue repair as a decisive determinant of the final outcome of liver injury inflicted by hepatotoxicants. These findings led us to propose a two-stage model of toxicity. In this model, tissue injury is inflicted in stage one by the well-described mechanisms of toxicity, whereas in stage two the ultimate toxic outcome is determined by whether timely and sufficient tissue repair response accompanies this injury. In an attempt to validate this model, dose-response relationships for injury and tissue repair as opposing responses have been developed for model hepatotoxicants. Results of these studies suggest that tissue repair increases in a dose-dependent manner, restraining injury up to a threshold dose, whereupon it is inhibited, allowing an unrestrained progression of injury. These findings indicate that tissue repair is a quantifiable response to toxic injury and that inclusion of this response in risk assessment may help in fine-tuning prediction of toxicity outcomes. PMID- 9860889 TI - Medium-term bioassays for carcinogenicity of chemical mixtures. AB - Carcinogenic effects of chemical mixtures were examined with a medium-term liver bioassay for carcinogens or a multiorgan medium-term bioassay using male F344 rats. In the medium-term liver bioassay, rats were initially treated with diethylnitrosamine (DEN) at 200 mg/kg body weight, i.p.; after 2 weeks they received chemical mixtures such as 10 different heterocyclic amines at one-tenth or one-hundredth the dose levels used in carcinogenicity studies and the mixtures of 20 different pesticides, each at acceptable daily intake (ADI) levels or a mixture of 100 times ADI levels. All animals were subjected to two-thirds partial hepatectomy at week 3 and were sacrificed at week 8. The number and areas of glutathione S-transferase placental form (GST-P) positive foci (preneoplastic lesions in the liver) were compared between respective groups. When 10 heterocyclic amines were mixed in the diet at one-tenth dose level, clear synergism was observed, but no combined effects were evident with the one hundredth dose levels. In the pesticide experiment, treatment of rats with the 20 pesticide mixture at the ADI dose level did not enhance GST-P-positive foci. In contrast, a mixture of 100 times the ADI significantly increased those values. In a multiorgan bioassay of 28 weeks, mixtures of 40 high-volume compounds and 20 pesticides (suspected carcinogens) added together at their respective ADI levels did not enhance carcinogenesis in any organs initiated by five different carcinogens (DEN, N-methylnitrosourea, dimethylhydrazine, N-butyl-N-(4 hydroxybutyl)nitrosamine, and dihydroxy-di-n-propylnitrosamine) in combination. The combination effect of low dietary levels of five antioxidants, butylated hydroxyanisole, caffeic acid, sesamol, 4-methoxyphenol, and catechol, were also examined using the multiorgan bioassay. The incidence of forestomach papillomas was significantly increased only in the combination group and the results indicate that combination of the five antioxidants can exert additive/synergistic effects on tumorigenesis in the multiorgan bioassay. These results indicate that chemical mixtures at very low doses did not enhance preneoplastic lesions synergistically but the mixtures at certain doses show synergism in the target organ. The medium-term bioassays are particularly useful tools for this purpose. PMID- 9860888 TI - Tumors and DNA adducts in mice exposed to benzo[a]pyrene and coal tars: implications for risk assessment. AB - Current methods to estimate the quantitative cancer risk of complex mixtures of polycyclic aromatic hydrocarbons (PAH) such as coal tar assume that overall potency can be derived from knowledge of the concentration of a few carcinogenic components such as benzo[a]pyrene (B[a]P). Genotoxic damage, such as DNA adducts, is thought to be an essential aspect of PAH-induced tumorigenesis and could be a biomarker for exposure useful for estimating risk. However, the role of B[a]P and the relationship of adduct formation in tumorigenesis have not been tested rigorously in models appropriate for human health risk assessment. Therefore, we directly compared tumor induction and adduct formation by B[a]P and coal tars in several experimental protocols, including one broadly accepted and used by regulators. We found that B[a]P content did not account for tumor incidences after exposure to coal tars. DNA adducts were found in both tumors and tumor-free tissue and tumor outcomes were not predicted by either quantitation of total DNA adducts or by the DNA adduct formed by B[a]P. These data suggest that risk assessments based on B[a]P content may not predict accurately risk to human health posed by environmental PAH. PMID- 9860890 TI - Lung tumorigenic interactions in strain A/J mice of five environmental polycyclic aromatic hydrocarbons. AB - The binary, ternary, quaternary, and quintary interactions of a five-component mixture of carcinogenic environmental polycyclic aromatic hydrocarbons (PAHs) using response surface analyses are described. Initially, lung tumor dose response curves in strain A/J mice for each of the individual PAHs benzo[a]pyrene (B[a]P), benzo[b]fluoranthene (B[b]F), dibenz[a,h]anthracene (DBA), 5 methylchrysene (5MC), and cyclopenta[cd]pyrene (CPP) were obtained. From these data, doses were selected for the quintary mixture study based on toxicity, survival, range of response, and predicted tumor yields. The ratios of doses among PAHs were designed to simulate PAH ratios found in environmental air and combustion samples. Quintary mixtures of B[a]P, B[b]F, DBA, 5MC, and CPP were administered to male strain A/J mice in a 2(5) factorial 32-dose group dosing scheme (combinations of five PAHs each at either high or low doses) and lung adenomas were scored. Comparison of observed lung adenoma formation with that expected from additivity identified both greater than additive and less than additive interactions that were dose related i.e., greater than additive at lower doses and less than additive at higher doses. To identify specific interactions, a response surface analysis using response addition was applied to the tumor data. This response surface model contained five dose, ten binary, ten ternary, five quaternary, and one quintary parameter. This analysis produced statistically significant values of 16 parameters. The model and model parameters were evaluated by estimating the dose-response relationships for each of the five PAHs. The predicted dose-response curves for all five PAHs indicated a good estimation. The binary interaction functions were dominated for the most part by DBA and were inhibitory. The response surface model predicted, to a significant degree, the observed lung tumorigenic responses of the quintary mixtures. These data suggest that although interactions between PAHs do occur, they are limited in extent. PMID- 9860893 TI - Statistically designed experiments to screen chemical mixtures for possible interactions. AB - For the accurate analysis of possible interactive effects of chemicals in a defined mixture, statistical designs are necessary to develop clear and manageable experiments. For instance, factorial designs have been successfully used to detect two-factor interactions. Particularly useful for this purpose are fractionated factorial designs, requiring only a fraction of all possible combinations of a full factorial design. Once the potential interaction has been detected with a fractionated design, a more accurate analysis can be performed for the particular binary mixtures to ensure and characterize these interactions. In this paper this approach is illustrated using an in vitro cytotoxicity assay to detect the presence of mixtures of Fusarium mycotoxins in contaminated food samples. We have investigated interactions between five mycotoxin species (Trichothecenes, Fumonisins, and Zearalenone) using the DNA synthesis inhibition assay in L929 fibroblasts. First, a central composite design was applied to identify possible interactive effects between mycotoxins in the mixtures (27 combinations from 5(5) possible combinations). Then two-factor interactions of particular interest were further analyzed by the use of a full factorial design (5 x 5 design) to characterize the nature of those interactions more precisely. Results show that combined exposure to several classes of mycotoxins generally results in effect addition with a few minor exceptions indicating synergistic interactions. In general, the nature of the interactions characterized in the full factorial design was similar to the nature of those observed in the central composite design. However, the magnitude of interaction was relatively small in the full factorial design. PMID- 9860892 TI - Estimation of toxicity of chemical mixtures through modeling of chemical interactions. AB - The Agency for Toxic Substances and Disease Registry (ATSDR), in collaboration with the Dutch Organization for Applied Scientific Research (TNO) Nutrition and Food Research Institute, is conducting studies to evaluate the role of chemical interactions in the expression of toxicity from low-level exposure to combinations of chemicals. The goal of this collaborative effort is to use a weight-of-evidence (WOE) approach to estimate joint toxicity of some simple chemical mixtures and to compare the estimations with test results from animal toxicity studies. The WOE approach uses individual chemical dose-response assessments and algorithms that incorporate various assumptions regarding potential chemical interactions. Qualitative evaluations were prepared for binary combinations of chemicals for the effect of butyl hydroxyanisole on di(2 ethylhexyl)phthalate, the effect of stannous chloride on Cd chloride (CdCl2), and the effect of CdCl2 on loperamide. Analyses of these evaluations and their comparison with the conclusions of laboratory animal experiments indicate that the WOE approach can be used to estimate qualitatively the joint toxicity of such simple mixtures. To further test the utility of the WOE approach, qualitative and semiquantitative evaluations were prepared for two chemical mixtures--one with similarly acting halogenated aliphatics (trichloroethylene, tetrachloroethylene, hexachloro-1,3-butadiene[HCBD], and 1,1,2-trichloro-3,3,3-trifluoropropene [TCTFP]) and the other with dissimilarly acting nephrotoxic components (mercuric chloride, lysinolalanine, D-limonene, and HCBD). These two sets of data were used to estimate the overall toxicities of the mixtures using the WOE algorithm for the mixture. The comparison of the results of the estimated toxicity with experimentally determined toxicity of the mixture of similarly acting nephrotoxicants demonstrated that the WOE approach correctly adjusted for the observed interactions in experimental animal studies. However, this was not true for the mixture of dissimilarly acting nephrotoxicants. This could be attributed to the fact that WOE evaluations are based on dose additivity that postulates that all chemicals in a given mixture act in the same way--by the same mechanism- and differ only in their potencies. In these cases the WOE approach evaluations, based on consideration of common mechanisms for simple chemical mixtures, can lead to better estimates of joint toxicity of chemical mixtures than the default assumption of dose additivity. The results also show that the WOE evaluations should be target-organ specific because none of the models tested could approximate the observed responses in organs other than the target organs in the laboratory animal studies. PMID- 9860891 TI - Comparative estrogenic activity of wine extracts and organochlorine pesticide residues in food. AB - The human diet contains industrial-derived, endocrine-active chemicals and higher levels of naturally occurring compounds that modulate multiple endocrine pathways. Hazard and risk assessment of these mixtures is complicated by noadditive interactions between different endocrine-mediated responses. This study focused on estrogenic chemicals in the diet and compared the relative potencies or estrogen equivalents (EQs) of the daily consumption of xenoestrogenic organochlorine pesticides in food (2.44 micrograms/day) with the EQs in a single 200-ml glass of red cabernet wine. The reconstituted organochlorine mixture contained 1,1,1-trichloro-2-(p-chlorophenyl)-2-(o chlorophenyl)ethane, 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane, 1,1-dichloro 2,2-bis(p-chlorophenyl)ethylene, endosulfan-1, endosulfan-2, p,p'-methoxychlor, and toxaphene; the relative proportion of each chemical in the mixture resembled the composition reported in a recent U.S. Food and Drug Administration market basket survey. The following battery of in vitro 17 beta-estradiol (E2) responsive bioassays were utilized in this study: competitive binding to mouse uterine estrogen receptor (ER); proliferation in T47D human breast cancer cells; luciferase (Luc) induction in human HepG2 cells transiently cotransfected with C3 Luc and the human ER, rat ER-alpha, or rat ER-beta; induction of chloramphenicol acetyltransferase (CAT) activity in MCF-7 human breast cancer cells transfected with E2-responsive cathepsin D-CAT or creatine kinase B-CAT plasmids. For these seven in vitro assays, the calculated EQs in extracts from 200 ml of red cabernet wine varied from 0.15 to 3.68 micrograms/day. In contrast, EQs for consumption of organochlorine pesticides (2.44 micrograms/day) varied from nondetectable to 1.24 ng/day. Based on results of the in vitro bioassays, organochlorine pesticides in food contribute minimally to dietary EQ intake. PMID- 9860894 TI - A nonlinear isobologram model with Box-Cox transformation to both sides for chemical mixtures. AB - The linear logistical isobologram is a commonly used and powerful graphical and statistical tool for analyzing the combined effects of simple chemical mixtures. In this paper a nonlinear isobologram model is proposed to analyze the joint action of chemical mixtures for quantitative dose-response relationships. This nonlinear isobologram model incorporates two additional new parameters, Ymin and Ymax, to facilitate analysis of response data that are not constrained between 0 and 1, where parameters Ymin and Ymax represent the minimal and the maximal observed toxic response. This nonlinear isobologram model for binary mixtures can be expressed as [formula: see text] In addition, a Box-Cox transformation to both sides is introduced to improve the goodness of fit and to provide a more robust model for achieving homogeneity and normality of the residuals. Finally, a confidence band is proposed for selected isobols, e.g., the median effective dose, to facilitate graphical and statistical analysis of the isobologram. The versatility of this approach is demonstrated using published data describing the toxicity of the binary mixtures of citrinin and ochratoxin as well as a new experimental data from our laboratory for mixtures of mercury and cadmium. PMID- 9860895 TI - Mixture design and multivariate analysis in mixture research. AB - Mixture design has been used to identify possible interactions between mutagens in a mixture. In this paper the use of mixture design in multidimensional isobolographic studies is introduced. Mutagenicity of individual nitro-polycyclic aromatic hydrocarbons (PAH) was evaluated is an organic extract of diesel exhaust particles (DEPs). The particles were extracted with dichloromethane (DCM). After replacing DCM with dimethyl sulfoxide, the extract was spiked with three individual nitro-PAH: 1-nitropyrene, 2-nitrofluorene, and 1,8-dinitropyrene. The nitro-PAH were added separately and in various combinations to the extract to determine the effects of each variable and to identify possible interactions between the individual nitro-PAH and between the nitro-PAH and the extract. The composition of the mixtures was determined by mixture design (linear axial normal) with four variables (the DEP extract and the three nitro-PAH, giving 8 different mixtures plus a triplicate centerpoint, i.e., a total of 11. The design supports a model with linear and interaction (product) terms. Two different approaches were used: traditional mixture design within a well-defined range on the linear part of the dose-response curves and an isobolographic mixture design with equipotent doses of each variable. The mixtures were tested for mutagenicity in the Ames assay using the TA98 strain of Salmonella typhimurium. The data were analyzed with projections to latent structures (PLS). The three individual nitro PAH and the DEP extract acted additively in the Ames test. The use of mixture design either within a well-defined range of the linear part on the dose-response curve or with equipotent doses saves experiments and reduces the possibility of false interaction terms in situations with dose additivity or response additivity. PMID- 9860896 TI - Physiological modeling of toxicokinetic interactions: implications for mixture risk assessment. AB - Most of the available data on chemical interactions have been obtained in animal studies conducted by administering high doses of chemicals by routes and scenarios different from anticipated human exposures. A mechanistic approach potentially useful for conducting dose, scenario, species, and route extrapolations of toxic interactions is physiological modeling. This approach involves the development of mathematical descriptions of the interrelationships among the critical determinants of toxicokinetics and toxicodynamics. The mechanistic basis of the physiological modeling approach not only enables the species, dose, route, and scenario extrapolations of the occurrence of toxicokinetic interactions but also allows the extrapolation of the occurrence of interactions from binary to multichemical mixtures. Examples are presented to show the feasibility of predicting changes in toxicokinetics of the components of complex chemical mixtures based on the incorporation of binary interaction data within physiologically based models. Interactions-based mixture risk assessment can be performed by simulating the change in the tissue dose of the toxic moiety of each mixture component during combined exposures and calculating the risk associated with each tissue dose estimate using a tissue dose versus response curve for all components. The use of such a mechanistic approach should facilitate the evaluation of the magnitude and relevance of chemical interactions in assessing the risks of low-level human exposures to complex chemical mixtures. PMID- 9860897 TI - Approaches to developing alternative and predictive toxicology based on PBPK/PD and QSAR modeling. AB - Systematic toxicity testing, using conventional toxicology methodologies, of single chemicals and chemical mixtures is highly impractical because of the immense numbers of chemicals and chemical mixtures involved and the limited scientific resources. Therefore, the development of unconventional, efficient, and predictive toxicology methods is imperative. Using carcinogenicity as an end point, we present approaches for developing predictive tools for toxicologic evaluation of chemicals and chemical mixtures relevant to environmental contamination. Central to the approaches presented is the integration of physiologically based pharmacokinetic/pharmacodynamic (PBPK/PD) and quantitative structure--activity relationship (QSAR) modeling with focused mechanistically based experimental toxicology. In this development, molecular and cellular biomarkers critical to the carcinogenesis process are evaluated quantitatively between different chemicals and/or chemical mixtures. Examples presented include the integration of PBPK/PD and QSAR modeling with a time-course medium-term liver foci assay, molecular biology and cell proliferation studies. Fourier transform infrared spectroscopic analyses of DNA changes, and cancer modeling to assess and attempt to predict the carcinogenicity of the series of 12 chlorobenzene isomers. Also presented is an ongoing effort to develop and apply a similar approach to chemical mixtures using in vitro cell culture (Syrian hamster embryo cell transformation assay and human keratinocytes) methodologies and in vivo studies. The promise and pitfalls of these developments are elaborated. When successfully applied, these approaches may greatly reduce animal usage, personnel, resources, and time required to evaluate the carcinogenicity of chemicals and chemical mixtures. PMID- 9860899 TI - Altered biologic activities of commercial polychlorinated biphenyl mixtures after microbial reductive dechlorination. AB - The reductive dechlorination of polychlorinated biphenyls (PCBs) by anaerobic bacteria has recently been established as an important environmental fate of these compounds. This process removes chlorines directly from the biphenyl ring with replacement by hydrogen, resulting in a product mixture in which the average number of chlorines per biphenyl is reduced. In this study, dechlorination of commercial PCB mixtures (Aroclors 1242 and 1254) by microorganisms eluted from PCB-contaminated sediments of the River Raisin (Michigan) and Silver Lake (Massachusetts) caused a depletion in the proportion of highly chlorinated PCB congeners and an accumulation of lesser-chlorinated congeners. Dechlorination occurred primarily at the meta and, to a much lesser extent, para positions of biphenyl. The concentrations of the coplanar congeners including 3,3',4,4',5 pentachlorobiphenyl, the most potent dioxinlike congener, were significantly lowered by reductive dechlorination. Microbial reductive dechlorination of commercial PCB mixtures caused a substantial reduction in biologic activities in several instances. It significantly lowered or eliminated the inhibitory effects of Aroclors on fertilization of mouse gametes in vitro. Similarly, the dechlorinated product mixtures had substantially lower ethoxyresorufin-O deethylase induction potencies and showed less ability to induce activating protein 1 transcription factor activity as compared to the unaltered Aroclors. In other assays the same dechlorinated product mixtures demonstrated biologic activities similar to the nondechlorinated Aroclors, including the ability of PCB mixtures to stimulate insulin secretion and cause neutrophil activation. The data presented here establish that the biologic activities of commercial PCB mixtures are altered by microbial reductive dechlorination and that an assessment of their toxic potential requires an array of tests that include the different mechanisms associated with PCBs. PMID- 9860900 TI - Using metal-ligand binding characteristics to predict metal toxicity: quantitative ion character-activity relationships (QICARs). AB - Ecological risk assessment can be enhanced with predictive models for metal toxicity. Modelings of published data were done under the simplifying assumption that intermetal trends in toxicity reflect relative metal-ligand complex stabilities. This idea has been invoked successfully since 1904 but has yet to be applied widely in quantitative ecotoxicology. Intermetal trends in toxicity were successfully modeled with ion characteristics reflecting metal binding to ligands for a wide range of effects. Most models were useful for predictive purposes based on an F-ratio criterion and cross-validation, but anomalous predictions did occur if speciation was ignored. In general, models for metals with the same valence (i.e., divalent metals) were better than those combining mono-, di-, and trivalent metals. The softness parameter (sigma p) and the absolute value of the log of the first hydrolysis constant ([symbol: see text] log KOH [symbol: see text]) were especially useful in model construction. Also, delta E0 contributed substantially to several of the two-variable models. In contrast, quantitative attempts to predict metal interactions in binary mixtures based on metal-ligand complex stabilities were not successful. PMID- 9860898 TI - Molecular and genetic ecotoxicologic approaches to aquatic environmental bioreporting. AB - Molecular and population genetic ecotoxicologic approaches are being developed for the utilization of arthropods as bioreporters of heavy metal mixtures in the environment. The explosion of knowledge in molecular biology, molecular genetics, and biotechnology provides an unparalleled opportunity to use arthropods as bioreporter organisms. Interspecific differences in aquatic arthropod populations have been previously demonstrated in response to heavy metal insult in the Arkansas River (AR) California Gulch Superfund site (CGSS). Population genetic analyses were conducted on the mayfly Baetis tricaudatus. Genetic polymorphisms were detected in polymerase chain reaction amplified 16S mitochondrial rDNA (a selectively neutral gene) of B tricaudatus using single-strand conformation polymorphism analysis. Genetic differences may have resulted from impediments to gene flow in the population caused by mortality arising from exposure to heavy metal mixture pollution. In laboratory studies a candidate metal-responsive mucinlike gene, which is metal and dose specific, has been identified in Chironomus tentans and other potential AR-CGSS bioreporter species. Population genetic analyses using the mucinlike gene may provide insight into the role of this selectable gene in determining the breeding structure of B. tricaudatus in the AR-CGSS and may provide mechanistic insight into determinants of aquatic arthropod response to heavy metal insult. Metal-responsive (MR) genes and regulatory sequences are being isolated, characterized, and assayed for differential gene expression in response to heavy metal mixture pollution in the AR-CGSS. Identified promoter sequences can then be engineered into previously developed MR constructs to provide sensitive in vitro assays for environmental bioreporting of heavy metal mixtures. The results of the population genetic studies are being entered into an AR geographic information system that contains substantial biological, chemical, and geophysical information. Integrated spatial, structural, and temporal analyses of these parameters will provide invaluable information concerning environmental determinants that restrict or promote gene flow in bioreporter populations. PMID- 9860901 TI - Genotoxicity of bioremediated soils from the Reilly Tar site, St. Louis Park, Minnesota. AB - An in vitro approach was used to measure the genotoxicity of creosote contaminated soil before and after four bioremediation processes. The soil was taken from the Reilly Tar site, a closed Superfund site in Saint Louis Park, Minnesota. The creosote soil was bioremediated in bioslurry, biopile, compost, and land treatment, which were optimized for effective treatment. Mutagenicity profiles of dichloromethane extracts of the five soils were determined in the Spiral technique of the Salmonella assay with seven tester strains. Quantitative mutagenic responses in the plate incorporation technique were then determined in the most sensitive strains, YG1041 and YG1042. Mutagenic potency (revertants per microgram extract) in YG1041 suggested that compost, land treatment, and untreated creosote soil extracts were moderately mutagenic with Arochlor-induced rat liver (S9) but were nonmutagenic without S9. However, the bioslurry extract was strongly mutagenic and the biopile extract was moderately mutagenic either with or without S9. A similar trend was obtained in strain YG1042. The strong mutagenic activity in the bioslurry extract was reduced by 50% in TA98NR, which suggested the presence of mutagenic nitrohydrocarbons. Variation in reproducibility was 15% or less for the bioassay and extraction procedures. Bioavailability of mutagens in the biopile soil was determined with six solvents; water-soluble mutagens accounted for 40% of the total mutagenic activity and they were stable at room temperature. The mutagenic activity in the bioslurry and biopsile samples was due to either the processes themselves or to the added sludge/manure amendments. The in vitro approach was effective in monitoring bioremediated soils for genotoxicity and will be useful in future laboratory and in situ studies. PMID- 9860903 TI - Modeling the chemistry of complex petroleum mixtures. AB - Determining the complete molecular composition of petroleum and its refined products is not feasible with current analytical techniques because of the astronomical number of molecular components. Modeling the composition and behavior of such complex mixtures in refinery processes has accordingly evolved along a simplifying concept called lumping. Lumping reduces the complexity of the problem to a manageable form by grouping the entire set of molecular components into a handful of lumps. This traditional approach does not have a molecular basis and therefore excludes important aspects of process chemistry and molecular property fundamentals from the model's formulation. A new approach called structure-oriented lumping has been developed to model the composition and chemistry of complex mixtures at a molecular level. The central concept is to represent an individual molecular or a set of closely related isomers as a mathematical construct of certain specific and repeating structural groups. A complex mixture such as petroleum can then be represented as thousands of distinct molecular components, each having a mathematical identity. This enables the automated construction of large complex reaction networks with tens of thousands of specific reactions for simulating the chemistry of complex mixtures. Further, the method provides a convenient framework for incorporating molecular physical property correlations, existing group contribution methods, molecular thermodynamic properties, and the structure--activity relationships of chemical kinetics in the development of models. PMID- 9860902 TI - Bioassay-directed fractionation and chemical identification of mutagens in bioremediated soils. AB - Soil from a Superfund site (Reilly Tar Site, St. Louis Park, Minnesota) contaminated with polycyclic aromatic hydrocarbons (PAHs) from creosote was treated with several bioremediation technologies including bioslurry (BS), biopile (BP), compost (CMP), and land treatment (LT). These treatment technologies are being evaluated in pilot scale laboratory systems by the U.S. Environmental Protection Agency's National Risk Management Research Laboratory in Cincinnati, Ohio. To evaluate the genotoxicity and identify the mutagens in the soil before and after the various treatments, fractionated extracts of five soils were bioassayed for mutagenic activity with a microsuspension modification of the Salmonella histidine reversion assay. Soils were extracted by sonication using dichloromethane (DCM). The five extracts were fractionated in triplicate (two for bioassay and one for chemical analysis) by reverse-phase high-performance liquid chromatography (HPLC) using hexane/DCM/methanol, and the fraction for bioassay were solvent-exchanged into dimethyl sulfoxide by nitrogen evaporation. Forty HPLC fractions for each sample were bioassayed in strain YG1041 with and without exogenous liver metabolic activation. As shown in a companion paper, the mutagenicity of two treatments (BS and BP) was significantly greater than the mutagenicity of the untreated soil. Mutagenic fractions (> 500 revertants) were analyzed by gas chromatography/mass spectrometry (GC/MS). PAH analysis of the soils indicated that all treatments were effective in reducing the total PAH concentration (48-74%). Qualitative GC/MS analysis of the mutagenic fractions from the BS and BP treatments indicated that they contained azaarenes, which are mutagens. The CMP and LT processes were the most effective and least toxic bioremediation procedures based on mutagenic potency and chemical analysis. This research demonstrated that the combination of bioassays and chemical analysis provided a more accurate determination of toxicity in these complex environmental mixtures. PMID- 9860904 TI - Evaluation (not validation) of quantitative models. AB - The present regulatory climate has led to increasing demands for scientists to attest to the predictive reliability of numerical simulation models used to help set public policy, a process frequently referred to as model validation. But while model validation may reveal useful information, this paper argues that it is not possible to demonstrate the predictive reliability of any model of a complex natural system in advance of its actual use. All models embed uncertainties, and these uncertainties can and frequently do undermine predictive reliability. In the case of lead in the environment, we may categorize model uncertainties as theoretical, empirical, parametrical, and temporal. Theoretical uncertainties are aspects of the system that are not fully understood, such as the biokinetic pathways of lead metabolism. Empirical uncertainties are aspects of the system that are difficult (or impossible) to measure, such as actual lead ingestion by an individual child. Parametrical uncertainties arise when complexities in the system are simplified to provide manageable model input, such as representing longitudinal lead exposure by cross-sectional measurements. Temporal uncertainties arise from the assumption that systems are stable in time. A model may also be conceptually flawed. The Ptolemaic system of astronomy is a historical example of a model that was empirically adequate but based on a wrong conceptualization. Yet had it been computerized--and had the word then existed- its users would have had every right to call it validated. Thus, rather than talking about strategies for validation, we should be talking about means of evaluation. That is not to say that language alone will solve our problems or that the problems of model evaluation are primarily linguistic. The uncertainties inherent in large, complex models will not go away simply because we change the way we talk about them. But this is precisely the point: calling a model validated does not make it valid. Modelers and policymakers must continue to work toward finding effective ways to evaluate and judge the quality of their models, and to develop appropriate terminology to communicate these judgments to the public whose health and safety may be at stake. PMID- 9860905 TI - Historical perspective on lead biokinetic models. AB - A historical review of the development of biokinetic model of lead is presented. Biokinetics is interpreted narrowly to mean only physiologic processes happening within the body. Proceeding chronologically, for each epoch, the measurements of lead in the body are presented along with mathematical models in an attempt to trace the convergence of observations from two disparate fields--occupational medicine and radiologic health--into some unified models. Kehoe's early balance studies and the use of radioactive lead tracers are presented. The 1960s saw the joint application of radioactive lead techniques and simple compartmental kinetic models used to establish the exchange rates and residence times of lead in body pools. The applications of stable isotopes to questions of the magnitudes of respired and ingested inputs required the development of a simple three-pool model. During the 1980s more elaborate models were developed. One of their key goals was the establishment of the dose-response relationship between exposure to lead and biologic precursors of adverse health effects. PMID- 9860906 TI - Uses and limits of empirical data in measuring and modeling human lead exposure. AB - This paper examines the uses and limits of empirical data in evaluating measurement and modeling approaches to human lead exposure. Empirical data from experiment or observation or both have been used in studies of lead exposure. For example, experimental studies have elucidated and quantified physiologic or biokinetic parameters of lead exposure under controlled conditions. Observation, i.e., epidemiology, has been widely applied to study population exposures to lead. There is growing interest in the use of lead exposure prediction models and their evaluation before use in risk assessment. Empirical studies of lead exposure must be fully understood, especially their limits, before they are applied as "standards" or reference information for evaluation of exposure models, especially the U.S. Environmental Protection Agency's lead biokinetic model that is a focus of this article. Empirical and modeled datasets for lead exposure may not agree due to a) problems with the observational data or b) problems with the model; caution should be exercised before either a model or observational data are rejected. There are at least three sources of discordance in cases where there is lack of agreement: a) empirical data are accurate but the model is flawed; b) the model is valid but reference empirical data are inaccurate; or c) neither empirical data nor model is accurate, and each is inaccurate in different ways. This paper evaluates some of the critical empirical input to biokinetic models, especially lead bioavailability. PMID- 9860907 TI - Predicting blood lead concentrations from lead in environmental media. AB - Policy statements providing health and environmental criteria for blood lead (PbB) often give recommendations on an acceptable distribution of PbB concentrations. Such statements may recommend distributions of PbB concentrations including an upper range (e.g., maximum and/or 90th percentile values) and central tendency (e.g., mean and/or 50th percentile) of the PbB distribution. Two major, and fundamentally dissimilar, methods to predict the distribution of PbB are currently in use: statistical analyses of epidemiologic data, and application of biokinetic models to environmental lead measurements to predict PbB. Although biokinetic models may include a parameter to predict contribution of lead from bone (PbBone), contemporary data based on chemical analyses of pediatric bone samples are rare. Dramatic decreases in environmental lead exposures over the past 15 years make questionable use of earlier data on PbBone concentrations to estimate a contribution of lead from bone; often used by physiologic modelers to predict PbB. X-ray fluorescent techniques estimating PbBone typically have an instrument-based quantitation limit that is too high for use with many young children. While these quantitation limits have improved during the late 1990s, PbBone estimates using an epidemiologic approach to describing these limits for general populations of children may generate values lower than the instrument's quantitation limit. Additional problems that occur if predicting PbB from environmental lead by biokinetic modeling include a) uncertainty regarding the fractional lead absorption by young children; b) questions of bioavailability of specific environmental sources of lead; and c) variability in fractional absorption values over a range of exposures. Additional sources of variability in lead exposures that affect predictions of PbB from models include differences in the prevalence of such child behaviors as intensity of hand-to-mouth activity and pica. In contrast with these sources of uncertainty and variability affecting physiologic modeling of PbB distributions, epidemiologic data reporting PbB values obtained by chemical analyses of blood samples avoid these problems but raise other issues about the validity of the representation of the subsample for the overall population of concern. State and local health department screening programs and/or medical evaluation of individual children provide PbB data that contribute to databases describing the impact of environmental sources on PbB. Overall, application of epidemiologic models involves fewer uncertainties and more readily reflects variability in PbB than does current state-of-the-art biokinetic modeling. PMID- 9860908 TI - A physiologically based kinetic model for lead in children and adults. AB - A physiologically based model of lead kinetics in children and adults has been developed and tested. The premises on which the physiologically based model is founded are reviewed in this paper. Because 95% or more of the body burden of lead in adults is found in the bone, bone metabolism is central to the model. Bone volumes are expressed as functions of body weight. Bone formation and resorption rates are estimated from human studies of stable labeled calcium kinetics. Cortical and trabecular bone are modeled separately, with their surface to-volume ratios taken into account. Standardized growth curves are used to relate body weight to age. Other model features such as organ volumes and physiologic functions are related to body weight based on measurements made in human subjects over a range of ages. Calibrations of the model to two human data sets are shown, and two applications to specific research questions are illustrated. A brief comparison of the structure of this model with that of the Leggett model, and a comparison of the output of this model with that of the integrated exposure uptake biokinetic model of the U.S. Environmental Protection Agency, are also included. PMID- 9860909 TI - The ICRP age-specific biokinetic model for lead: validations, empirical comparisons, and explorations. AB - The objective of this manuscript is to provide a description of the International Commission for Radiation Protection (ICRP) model and a comparison to other models (the integrated exposure uptake biokinetic [IEUBK] and O'Flaherty models), including the software used with the models, and a comparison of the model predictions for selected situations. The ICRP biokinetic model for Pb is a multicompartmental model for Pb uptake and disposition in children and in adults. The model describes deposition and retention of absorbed Pb in numerous tissues, removal from tissues to plasma, and movement along various routes of excretion. Long-term skeletal behavior of Pb is described in terms of age-specific rates of restructuring of compact and trabecular bone. The ICRP model is more flexible and has wider applicability than the IEUBK model. The major disadvantages are that application of the computer model requires some basic computer skills, and the user must convert the Pb concentrations in food, air, soil, dust, paint, or other media to the amount of Pb ingested or inhaled per day. Direct comparisons between the ICRP model and the IEUBK model are provided by modeling blood Pb levels using the IEUBK v0.99d default Pb uptakes and intake values. The model is used to simulate occupational exposure cases and a controlled Pb inhalation experiment in adult humans. Finally, use of the model to explore situations with limited data is illustrated by simulating the kinetics and disposition of Pb during acute Pb poisoning and chelation therapy in a child. PMID- 9860910 TI - The conceptual structure of the integrated exposure uptake biokinetic model for lead in children. AB - The integrated exposure uptake biokinetic model for lead in children was developed to provide plausible blood lead distributions corresponding to particular combinations of multimedia lead exposure. The model is based on a set of equations that convert lead exposure (expressed as micrograms per day) to blood lead concentration (expressed as micrograms per deciliter) by quantitatively mimicking the physiologic processes that determine blood lead concentration. The exposures from air, food, water, soil, and dust are modeled independently by several routes. Amounts of lead absorbed are modeled independently for air, food, water, and soil/dust, then combined as a single input to the blood plasma reservoir of the body. Lead in the blood plasma reservoir, which includes extracellular fluids, is mathematically allocated to all tissues of the body using age-specific biokinetic parameters. The model calculation provides the estimate for blood lead concentration for that age. This value is treated as the geometric mean of possible values for a single child, or the geometric mean of expected values for a population of children exposed to the same lead concentrations. The distribution of blood lead concentrations about this geometric mean is estimated using a geometric standard deviation, typically 1.6, derived from the analysis of well-conducted community blood studies. PMID- 9860911 TI - Structure, use, and validation of the IEUBK model. AB - The potential impact of the effects of lead in children is a major concern. Although measurements of lead concentration can be made in a geographic area, it is difficult to predict the effects of this exposure that involve complicated biologic functions. Dynamic mathematical models that can be simulated on a digital computer provide one method of analysis to facilitate the prediction process. The integrated exposure uptake biokinetic (IEUBK) model is a dynamic mathematical model that has been discretized for execution on a digital computer. This paper is concerned with the general difficulties in validating a dynamic model of this type. A number of the general pitfalls of validating a model of this type are presented. The illustrations are of a general nature not requiring an understanding of the physiologic effects of lead on children. The concept of validating a model by comparing results to historical data is discussed. A comparison is made with traditional modeling efforts having this form of dynamic model. Also included are general mathematic concepts illustrating potential difficulties with intuitive analyses in calibrating a dynamic model. PMID- 9860912 TI - Measurement error, biases, and the validation of complex models for blood lead levels in children. AB - Measurement error causes biases in regression fits. If one could accurately measure exposure to environmental lead media, the line obtained would differ in important ways from the line obtained when one measures exposure with error. The effects of measurement error vary from study to study. It is dangerous to take measurement error corrections derived from one study and apply them to data from entirely different studies or populations. Measurement error can falsely invalidate a correct (complex mechanistic) model. If one builds a model such as the integrated exposure uptake biokinetic model carefully, using essentially error-free lead exposure data, and applies this model in a different data set with error-prone exposures, the complex mechanistic model will almost certainly do a poor job of prediction, especially of extremes. Although mean blood lead levels from such a process may be accurately predicted, in most cases one would expect serious underestimates or overestimates of the proportion of the population whose blood lead level exceeds certain standards. PMID- 9860913 TI - Some useful statistical methods for model validation. AB - Although formal hypothesis tests provide a convenient framework for displaying the statistical results of empirical comparisons, standard tests should not be used without consideration of underlying measurement error structure. As part of the validation process, predictions of individual blood lead concentrations from models with site-specific input parameters are often compared with blood lead concentrations measured in field studies that also report lead concentrations in environmental media (soil, dust, water, paint) as surrogates for exposure. Measurements of these environmental media are subject to several sources of variability, including temporal and spatial sampling, sample preparation and chemical analysis, and data entry or recording. Adjustments for measurement error must be made before statistical tests can be used to empirically compare environmental data with model predictions. This report illustrates the effect of measurement error correction using a real dataset of child blood lead concentrations for an undisclosed midwestern community. We illustrate both the apparent failure of some standard regression tests and the success of adjustment of such tests for measurement error using the SIMEX (simulation-extrapolation) procedure. This procedure adds simulated measurement error to model predictions and then subtracts the total measurement error, analogous to the method of standard additions used by analytical chemists. PMID- 9860914 TI - The integrated exposure uptake biokinetic model for lead in children: independent validation and verification. AB - The U.S. Environmental Protection Agency employs a model, the integrated exposure biokinetic (IEUBK) model for lead in children, for the assessment of risks to children posed by environmental lead at hazardous waste sites. This paper describes results of an effort to verify the consistency of the documentation with the computer model and to test the computer code using a group that is independent from those involved in the model development. This review concluded that the IEUBK model correctly calculates the equations specified in the IEUBK model theory documentation. However, several issues were identified on model documentation, model performance, and the C++ programming language code (i.e., IEUBK model source code) documentation. These issues affect the ability of an independent reviewer to understand the workings of the IEUBK model but not the model's reliability. As a result of these findings, recommendations have been provided for updating documentation to the model as well as associated adjustments to the model documentation. PMID- 9860915 TI - Integrated exposure uptake biokinetic model for lead in children: empirical comparisons with epidemiologic data. AB - The concept of model validation is evolving in the scientific community. This paper addresses the comparison of observed and predicted estimates as one component of model validation as applied to the integrated exposure uptake biokinetic (IEUBK) model for lead in children. The IEUBK model is an exposure (dose)-response model that uses children's environmental lead exposures to estimate risk of elevated blood lead (typically > 10 micrograms/dl) through estimation of lead body burdens in a mass balance framework. We used residence specific environmental lead measurements from three epidemiologic datasets as inputs for the IEUBK model to predict blood lead levels, and compared these predictions with blood lead levels of children living at these residences. When the IEUBK modeling focused on children with representative exposure measurements, that is, children who spent the bulk of their time near the locations sampled, there was reasonably close agreement between observed and predicted blood lead distributions in the three studies considered. Geometric mean observed and predicted blood lead levels were within 0.7 microgram/dl, and proportions of study populations expected to be above 10 micrograms/dl were within 4% of those observed. PMID- 9860916 TI - Blood lead slope factor models for adults: comparisons of observations and predictions. AB - Here we explore the appropriateness of various parameter values for the Bowers et al. model [Risk Anal 14:183-189, 1994] in the context of predicting the influence of site-related exposure to lead in soil on the blood lead (PbB) levels of women of childbearing age. We outline the parameters prescribed by Bowers et al. as well as those prescribed by the U.S. Environmental Protection Agency (U.S. EPA). Comparison of the PbB levels predicted by the Bowers et al. model to those predicted by the validated O'Flaherty pharmacokinetic model indicates that the Bowers et al. model performs favorably when parameter values prescribed here are used. Use of the U.S. EPA-prescribed parameters yields predicted PbB levels that substantially exceed the validated O'Flaherty model predictions. Finally, both the U.S. EPA-prescribed parameter values and the parameter values recommended herein are used to predict PbB levels among adults living in four Superfund communities. Comparison of predicted PbB levels for these communities indicates that the U.S. EPA parameters overstate the incremental influence of lead in soil on PbB levels. Differences between the parameter values prescribed here and the U.S. EPA-prescribed parameters yield substantially different cleanup criteria for lead in soil, although conservative parameter values may still be appropriate for screening purposes. PMID- 9860917 TI - An empirical comparison of lead exposure pathway models. AB - Structural equation modeling is a statistical method for partitioning the variance in a set of interrelated multivariate outcomes into that which is due to direct, indirect, and covariate (exogenous) effects. Despite this model's flexibility to handle different experimental designs, postulation of a causal chain among the endogenous variables and the points of influence of the covariates is required. This has motivated the researchers at the University of Cincinnati Department of Environmental Health to be guided by a theoretical model for movement of lead from distal sources (exterior soil or dust and paint lead) to proximal sources (interior dust lead) and then finally to biologic outcomes (handwipe and blood lead). The question of whether a single structural equation model built from proximity arguments can be applied to diverse populations observed in different communities with varying lead amounts, sources, and bioavailabilities is addressed in this article. This reanalysis involved data from 1855 children less than 72 months of age enrolled in 11 studies performed over approximately 15 years. Data from children residing near former ore processing sites were included in this reanalysis. A single model adequately fit the data from these 11 studies; however, the model needs to be flexible to include pathways that are not frequently observed. As expected, the more proximal sources of interior dust lead and handwipe lead were the most important predictors of blood lead; soil lead often had a number of indirect influences. A limited number of covariates were also isolated as usually affecting the endogenous lead variables. The blood lead levels surveyed at the ore-processing sites were comparable to and actually somewhat lower than those reported in the the Third National Health and Nutrition Examination Survey. Lessened bioavailability of the lead at certain of these sites is a probable reason for this finding. PMID- 9860918 TI - Roles of lead-binding proteins in mediating lead bioavailability. AB - The intracellular bioavailability of lead (Pb) at low dosage levels in major target organs such as the kidney and brain appears to be largely determined by complexation with a group of low molecular weight proteins. These proteins are rich in aspartic and glutamic dicarboxyl amino acids. The proteins are chemically similar but not identical across all species examined to date and the brain protein appears to be different from that found in the kidney. These proteins possess dissociation constant values for Pb on the order of 10(-8) M and appear to normally bind zinc. In rats, these proteins attenuate the Pb inhibition of the heme pathway enzyme delta-aminolevulinic acid dehydratase by a mechanism involving both Pb chelation and zinc donation to this highly Pb-sensitive zinc dependent enzyme. Other studies in rats have shown that the kidney protein facilitates the intranuclear movement of Pb in vitro followed by chromatin binding, suggesting that this protein may be involved in alterations of the pathognomonic Pb intranuclear inclusion bodies in renal gene expression associated with the mitogenic effects of Pb in the kidney. PMID- 9860920 TI - Preface PMID- 9860921 TI - The 1996 Bernard B. Brodie lecture: A journey in cytochrome P450 and drug metabolism research. PMID- 9860919 TI - Bioavailability of soilborne lead in adults, by stable isotope dilution. AB - Using stable isotope dilution, we determined the bioavailability of soilborne lead (Pb) in human adult volunteers. Soil from a residential yard at a mining impacted federal Superfund site that had negligible amounts of other priority pollutants was dried and screened through a 25-micron mesh sieve. The < 250 micron fraction, which likely represents that ingested via hand-to-mouth activity, was then sterilized by exposure to radiation. Ten replicate samples yielded a mean (SD) soil Pb concentration of 2924 +/- 36 ppm, and a mean 206Pb/207Pb ratio of 1.1083 +/- 0.0002, indicating remarkable soil homogeneity. Six adults with 206Pb/207Pb ratios of > 1.190 were admitted to the clinical research center and fasted overnight prior to dosing with 250 micrograms Pb/70 kg bw (i.e., 85.5 mg soil/70 kg) in a gelatin capsule. Blood for Pb and 206Pb/207Pb ratios was obtained at 14 time points through 30 hr. Results of the isotopic analyses from these subjects indicate that on average 26.2% +/- 8.1 of the administered dose was absorbed. Six additional subjects were subsequently studied but ingested soil immediately after a standardized breakfast. Bioavailability in this group was only 2.52% +/- 1.7. Collectively, this study provides the first experimental estimates of soil Pb absorption in humans, and should allow for more precise estimates of health risks due to Pb-contaminated soil. PMID- 9860922 TI - The life and times of anthony Y. H. Lu PMID- 9860923 TI - Twenty years of biochemistry of human P450s: purification, expression, mechanism, and relevance to drugs. AB - Today cytochrome P450 (P450) research is accepted as an integral part of drug development and discovery. Work leading to this point included biochemical studies on P450 in experimental animal models and application to human systems. The development of recombinant expression systems has been an important part of the progress, and in this article we describe some recently developed bacterial systems that can be used for the production of metabolites, genotoxicity testing, and screening in random mutagenesis work. Rate-limiting aspects of P450 reactions vary with particular systems, and further investigations are in order. Non-ionic detergents have been utilized widely in P450 purification work; these compounds are now shown to be substrates for P450s. These oxidations are not only of fundamental interest in expanding the repertoire of P450 substrates but have significance in light of human exposure to these compounds. PMID- 9860924 TI - Molecular basis of P450 inhibition and activation: implications for drug development and drug therapy. AB - Three-dimensional homology models of cytochromes P450 (P450) 2B1 and P450 3A4 have been utilized along with site-directed mutagenesis to elucidate the molecular determinants of substrate specificity. Most of the key residues identified in 2B enzymes fall within five substrate recognition sites (SRSs) and have counterparts in bacterial P450 residues that regulate substrate binding or access. Docking of inhibitors into 2B models has provided a plausible explanation for changes in susceptibility to mechanism-based inactivation that accompany particular amino acid side-chain replacements. These studies provide a basis for predicting drug interactions due to P450 inhibition and for rational inhibitor design. In addition, the location of P450 3A4 residues capable of influencing homotropic stimulation by substrates and heterotropic stimulation by flavonoids has been identified. Steroid hydroxylation by the wild-type enzyme exhibits sigmoidal kinetics, indicative of positive cooperativity. Based on the 3A4 model and single-site mutants, a double mutant in SRS-2 has been constructed that exhibits normal Michaelis-Menten kinetics. Results of modeling and mutagenesis studies suggest that the substrate and effector bind at adjacent sites within a single large cavity in P450 3A4. A thorough understanding of the location and structural requirements of the substrate-binding and effector sites in cytochrome P450 3A4 should prove valuable in rationalizing and predicting interactions among the multitude of drugs and other compounds that bind to the enzyme. PMID- 9860925 TI - Nitric oxide synthase structure and electron transfer. AB - The nitric oxide synthases (NOS), although unrelated to the cytochromes P450 in terms of sequence, exhibit spectroscopic and catalytic properties strongly reminiscent of those of the P450 system. One important difference is the requirement of the NOS enzymes for tetrahydrobiopterin. The biopterin cofactor is shown by chemical studies to bind close to pyrrole ring D of the prosthetic heme group, a position confirmed recently for inducible NOS and endothelial NOS by crystal structures. The only plausible role so far for the tetrahydrobiopterin is as a transient electron donor for the activation of molecular oxygen. NADPH derived electrons are provided to the heme by the NOS flavin domain, but the biopterin may be required to provide an electron at a faster rate than that supported by the flavin groups. Chimeras in which the reductase domains of the isoforms have been exchanged indicate that the overall rate of catalytic turnover is directly governed by the ability of the flavin domain to deliver electrons. Electron transfer from the flavin to the heme domain, and within the flavin and heme domains, is thus a critical determinant of the catalytic turnover of NOS. PMID- 9860927 TI - The aryl hydrocarbon receptor: studies using the AHR-null mice. AB - The aryl hydrocarbon receptor (AHR) is believed to mediate the toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), polychlorinated biphenyls, and polycyclic aromatic hydrocarbons. AHR is a member of the Per, ARNT, Sim/basic helix-loop-helix superfamily of ligand-activated transcription factors that also harbors the transcription factors involved in the hypoxia response, development of the central nervous system, and day-night adaptations. To investigate the role of AHR in chemical toxicity and carcinogenesis and to determine any possible function in mammalian development and physiological homeostasis, AHR-null mice were developed. The AHR-null mice were resistant to the acute toxicity of TCDD and had an altered teratogenic response to this compound. These mice were found to have a number of abnormal phenotypes, thus confirming that AHR plays an important developmental and physiological role. Among the most consistent phenotypes was an altered liver pathology that was associated with accelerated rates of apoptosis. Evidence suggests that this may be related to an abnormal accumulation of levels of hepatic retinoic acid that cause an activation of transforming growth factor beta, resulting in stimulation of apoptosis. AHR may directly or indirectly control levels of a cytochrome P450 that is responsible for catabolizing retinoic acid. PMID- 9860926 TI - Multiple activated oxygen species in P450 catalysis: contributions To specificity in drug metabolism. AB - A hypervalent iron-oxene species has been widely proposed as the "active oxygen" in cytochrome P450 (P450)-catalyzed reactions. We recently examined the effect of mutation of the highly conserved threonine residue in P450s 2B4 and 2E1 to alanine, a change that is believed to interfere with proton delivery to the active site, and have determined the change in rates of deformylation of aldehydes, epoxidation of olefins, and hydroxylation of various substrates. The results support the concept that three distinct oxidants are functional in P450 catalysis: nucleophilic peroxo-iron, nucleophilic or electrophilic hydroperoxo iron, and electrophilic oxenoid-iron. The occurrence of multiple oxidizing species may contribute to the remarkable versatility of the P450 family of isozymes in the modification of drugs and other substrates. Furthermore, the relative concentrations of these oxidants in a particular P450 isozyme may contribute to substrate specificity and govern the type of reaction catalyzed. PMID- 9860929 TI - Applications and limitations of interspecies scaling and in vitro extrapolation in pharmacokinetics. AB - The search for new drugs is an extremely time-consuming and costly endeavor. Much of the time and cost are expended on generating data that support the efficacy and safety profiles of the drug. Because of ethical constraints, relevant pharmacological and toxicological assessments must be made in laboratory animals and in in vitro systems before human testing can begin. In support of the efficacy and safety evaluation during drug development, two fundamental challenges facing industrial drug metabolism scientists are (1) how to "scale-up" the pharmacokinetic data from animals to humans and (2) how to extrapolate the in vitro data to the in vivo situation. This review examines the applications and limitations of interspecies scaling and in vitro extrapolation in pharmacokinetics. PMID- 9860928 TI - Lanosterol 14alpha-demethylase (CYP51) and spermatogenesis. AB - CYP51 is the only gene of the cytochrome P450 (P450, or CYP) superfamily that is expressed in prokaryotes and eukaryotes. In animals, the gene product, P45014DM, catalyzes the lanosterol 14alpha-demethylase reaction, an essential step in cholesterol biosynthesis. P45014DM serves a housekeeping role, and it was surprising to find the highest level of CYP51 expression in the testes. This is a result of very high-level CYP51 expression in postmeiotic, haploid spermatids and results in elevated P45014DM activity in these cells. It is proposed that the elevated level of 14alpha-demethylase activity leads to production of signaling sterols by haploid germ cells, although the function of such sterols in males is unknown. PMID- 9860930 TI - Short- and long-term projections about the use of drug metabolism in drug discovery and development. AB - The science of drug metabolism, like any other science, has advanced from simple beginnings (by today's standards) to its present state. One can examine the path that has been taken to understand the forces driving the direction of evolution of this science. The trends discovered can then be used to make reasonable extrapolations about the changes that might be expected in the future. That exercise is the subject of this article. The main focus will be on drug metabolism as practiced in the industrial environment, representing the author's main experience as well as the principal arena of practical applications of the science. The discussion will draw mainly on broad phenomena occurring in this application of drug metabolism to drug discovery and development. PMID- 9860931 TI - Drug-metabolism research challenges in the new millennium: individual variability in drug therapy and drug safety. AB - One of the most challenging research areas in pharmacology in the new millennium is to understand why individuals respond differently to drug therapy and to what extent that individual variability in disposition is responsible for the observed differences in therapeutic efficacy and adverse reactions. To answer these complex questions, drug-metabolism research will rely on multidisciplinary approaches more than ever to investigate the many components involved in drug metabolism and disposition. Major research challenges include the following: (1) the genetic variation of drug targets (receptors, enzymes, etc.), drug transporters (multispecific organic anion transporter, P-glycoprotein, alpha-1 acid glycoprotein, etc.), and drug-metabolizing enzymes (cytochrome P450s and other enzymes); (2) the structure and function of all genetic variants of drug receptors, transporters, and metabolizing enzymes; (3) the induction, repression, and inhibition of all components involved in drug disposition; (4) the development of noninvasive in vivo methods to determine the physiological significance of various components in the handling of specific therapeutic agents in humans; (5) the mechanism of idiosyncratic adverse drug reactions; and (6) the pharmacokinetic and pharmacodynamic relationships to explain the individual differences in therapeutic efficacy and drug safety. Thus successful drug metabolism research in the new millennium must integrate receptor biology, enzymology, recombinant DNA technology, biochemical toxicology, and drug disposition into study design and conduct balanced in vitro and in vivo experiments to allow a full understanding of the mechanisms of individual variability in drug therapy and drug safety. PMID- 9860932 TI - Structure-function of cytochromes P450 and flavin-containing monooxygenases: implications for drug metabolism. AB - This article is a report on a symposium held at Experimental Biology '98 in San Francisco, California. Recent developments in site-directed mutagenesis, computer modeling, and mechanistic analysis of cytochromes P450 and flavin-containing monooxygenases are described. A unifying theme is the elaboration of general approaches for understanding and predicting the function of individual forms of these enzymes. A related goal is the production of soluble forms of mammalian cytochromes P450 for X-ray crystallography. PMID- 9860933 TI - Physiological and pathophysiological regulation of cytochrome P450. AB - This article is a report on a symposium sponsored by the American Society for Pharmacology and Experimental Therapeutics and held at the April 1998 Experimental Biology '98 meeting in San Francisco. The presentations focused on the mechanisms of regulation of cytochrome P450 gene expression by developmental factors and by hormones and cytokines, as well as on the interplay between physiological and chemical regulation. Approaches and systems used to address these questions included conditional gene knockouts in mice, primary hepatocyte cultures, immunofluorescence imaging of cells, and cell lines stably expressing reporter gene constructs. PMID- 9860934 TI - Serotonin receptor knockouts: a moody subject. PMID- 9860935 TI - Stem cell homing: rolling, crawling, and nesting. PMID- 9860937 TI - Protein dynamics and photon echoes. PMID- 9860936 TI - New insights into the tumor suppression function of P27(kip1) PMID- 9860938 TI - DNA curvature and deformation in protein-DNA complexes: a step in the right direction. PMID- 9860939 TI - Information coding in the cortex by independent or coordinated populations. PMID- 9860940 TI - Interaction between like-charged colloidal spheres in electrolyte solutions. AB - How colloidal particles interact with each other is one of the key issues that determines our ability to interpret experimental results for phase transitions in colloidal dispersions and our ability to apply colloid science to various industrial processes. The long-accepted theories for answering this question have been challenged by results from recent experiments. Herein we show from Monte Carlo simulations that there is a short-range attractive force between identical macroions in electrolyte solutions containing divalent counterions. Complementing some recent and related results by others, we present strong evidence of attraction between a pair of spherical macroions in the presence of added salt ions for the conditions where the interacting macroion pair is not affected by any other macroions that may be in the solution. This attractive force follows from the internal-energy contribution of counterion mediation. Contrary to conventional expectations, for charged macroions in an electrolyte solution, the entropic force is repulsive at most solution conditions because of localization of small ions in the vicinity of macroions. Both Derjaguin-Landau-Verwey-Overbeek theory and Sogami-Ise theory fail to describe the attractive interactions found in our simulations; the former predicts only repulsive interaction and the latter predicts a long-range attraction that is too weak and occurs at macroion separations that are too large. Our simulations provide fundamental "data" toward an improved theory for the potential of mean force as required for optimum design of new materials including those containing nanoparticles. PMID- 9860941 TI - Biochemical evolution II: origin of life in tubular microstructures on weathered feldspar surfaces. AB - Mineral surfaces were important during the emergence of life on Earth because the assembly of the necessary complex biomolecules by random collisions in dilute aqueous solutions is implausible. Most silicate mineral surfaces are hydrophilic and organophobic and unsuitable for catalytic reactions, but some silica-rich surfaces of partly dealuminated feldspars and zeolites are organophilic and potentially catalytic. Weathered alkali feldspar crystals from granitic rocks at Shap, north west England, contain abundant tubular etch pits, typically 0.4-0.6 microm wide, forming an orthogonal honeycomb network in a surface zone 50 microm thick, with 2-3 x 10(6) intersections per mm2 of crystal surface. Surviving metamorphic rocks demonstrate that granites and acidic surface water were present on the Earth's surface by approximately 3.8 Ga. By analogy with Shap granite, honeycombed feldspar has considerable potential as a natural catalytic surface for the start of biochemical evolution. Biomolecules should have become available by catalysis of amino acids, etc. The honeycomb would have provided access to various mineral inclusions in the feldspar, particularly apatite and oxides, which contain phosphorus and transition metals necessary for energetic life. The organized environment would have protected complex molecules from dispersion into dilute solutions, from hydrolysis, and from UV radiation. Sub-micrometer tubes in the honeycomb might have acted as rudimentary cell walls for proto-organisms, which ultimately evolved a lipid lid giving further shelter from the hostile outside environment. A lid would finally have become a complete cell wall permitting detachment and flotation in primordial "soup." Etch features on weathered alkali feldspar from Shap match the shape of overlying soil bacteria. PMID- 9860942 TI - Structure of a biological oxygen sensor: a new mechanism for heme-driven signal transduction. AB - The FixL proteins are biological oxygen sensors that restrict the expression of specific genes to hypoxic conditions. FixL's oxygen-detecting domain is a heme binding region that controls the activity of an attached histidine kinase. The FixL switch is regulated by binding of oxygen and other strong-field ligands. In the absence of bound ligand, the heme domain permits kinase activity. In the presence of bound ligand, this domain turns off kinase activity. Comparison of the structures of two forms of the Bradyrhizobium japonicum FixL heme domain, one in the "on" state without bound ligand and one in the "off" state with bound cyanide, reveals a mechanism of regulation by a heme that is distinct from the classical hemoglobin models. The close structural resemblance of the FixL heme domain to the photoactive yellow protein confirms the existence of a PAS structural motif but reveals the presence of an alternative regulatory gateway. PMID- 9860943 TI - Molecular recognition of angiogenesis inhibitors fumagillin and ovalicin by methionine aminopeptidase 2. AB - Angiogenesis inhibitors are a novel class of promising therapeutic agents for treating cancer and other human diseases. Fumagillin and ovalicin compose a class of structurally related natural products that potently inhibit angiogenesis by blocking endothelial cell proliferation. A synthetic analog of fumagillin, TNP 470, is currently undergoing clinical trials for treatment of a variety of cancers. A common target for fumagillin and ovalicin recently was identified as the type 2 methionine aminopeptidase (MetAP2). These natural products bind MetAP2 covalently, inhibiting its enzymatic activity. The specificity of this binding is underscored by the lack of inhibition of the closely related type 1 enzyme, MetAP1. The molecular basis of the high affinity and specificity of these inhibitors for MetAP2 has remained undiscovered. To determine the structural elements of these inhibitors and MetAP2 that are involved in this interaction, we synthesized fumagillin analogs in which each of the potentially reactive epoxide groups was removed either individually or in combination. We found that the ring epoxide in fumagillin is involved in the covalent modification of MetAP2, whereas the side chain epoxide group is dispensable. By using a fumagillin analog tagged with fluorescein, His-231 in MetAP2 was identified as the residue that is covalently modified by fumagillin. Site-directed mutagenesis of His-231 demonstrated its importance for the catalytic activity of MetAP2 and confirmed that the same residue is covalently modified by fumagillin. These results, in agreement with a recent structural study, suggest that fumagillin and ovalicin inhibit MetAP2 by irreversible blockage of the active site. PMID- 9860944 TI - Structure-based assignment of the biochemical function of a hypothetical protein: a test case of structural genomics. AB - Many small bacterial, archaebacterial, and eukaryotic genomes have been sequenced, and the larger eukaryotic genomes are predicted to be completely sequenced within the next decade. In all genomes sequenced to date, a large portion of these organisms' predicted protein coding regions encode polypeptides of unknown biochemical, biophysical, and/or cellular functions. Three-dimensional structures of these proteins may suggest biochemical or biophysical functions. Here we report the crystal structure of one such protein, MJ0577, from a hyperthermophile, Methanococcus jannaschii, at 1.7-A resolution. The structure contains a bound ATP, suggesting MJ0577 is an ATPase or an ATP-mediated molecular switch, which we confirm by biochemical experiments. Furthermore, the structure reveals different ATP binding motifs that are shared among many homologous hypothetical proteins in this family. This result indicates that structure-based assignment of molecular function is a viable approach for the large-scale biochemical assignment of proteins and for discovering new motifs, a basic premise of structural genomics. PMID- 9860945 TI - Structural code for DNA recognition revealed in crystal structures of papillomavirus E2-DNA targets. AB - Transcriptional regulation in papillomaviruses depends on sequence-specific binding of the regulatory protein E2 to several sites in the viral genome. Crystal structures of bovine papillomavirus E2 DNA targets reveal a conformational variant of B-DNA characterized by a roll-induced writhe and helical repeat of 10.5 bp per turn. A comparison between the free and the protein bound DNA demonstrates that the intrinsic structure of the DNA regions contacted directly by the protein and the deformability of the DNA region that is not contacted by the protein are critical for sequence-specific protein/DNA recognition and hence for gene-regulatory signals in the viral system. We show that the selection of dinucleotide or longer segments with appropriate conformational characteristics, when positioned at correct intervals along the DNA helix, can constitute a structural code for DNA recognition by regulatory proteins. This structural code facilitates the formation of a complementary protein-DNA interface that can be further specified by hydrogen bonds and nonpolar interactions between the protein amino acids and the DNA bases. PMID- 9860946 TI - Analysis of the regulatory phosphorylation site in Acanthamoeba myosin IC by using site-directed mutagenesis. AB - The actin-activated ATPase activity of Acanthamoeba myosin IC is stimulated 15- to 20-fold by phosphorylation of Ser-329 in the heavy chain. In most myosins, either glutamate or aspartate occupies this position, which lies within a surface loop that forms part of the actomyosin interface. To investigate the apparent need for a negative charge at this site, we mutated Ser-329 to alanine, asparagine, aspartate, or glutamate and coexpressed the Flag-tagged wild-type or mutant heavy chain and light chain in baculovirus-infected insect cells. Recombinant wild-type myosin IC was indistinguishable from myosin IC purified from Acanthamoeba as determined by (i) the dependence of its actin-activated ATPase activity on heavy-chain phosphorylation, (ii) the unusual triphasic dependence of its ATPase activity on the concentration of F-actin, (iii) its Km for ATP, and (iv) its ability to translocate actin filaments. The Ala and Asn mutants had the same low actin-activated ATPase activity as unphosphorylated wild type myosin IC. The Glu mutant, like the phosphorylated wild-type protein, was 16 fold more active than unphosphorylated wild type, and the Asp mutant was 8-fold more active. The wild-type and mutant proteins had the same Km for ATP. Unphosphorylated wild-type protein and the Ala and Asn mutants were unable to translocate actin filaments, whereas the Glu mutant translocated filaments at the same velocity, and the Asp mutant at 50% the velocity, as phosphorylated wild type proteins. These results demonstrate that an acidic amino acid can supply the negative charge in the surface loop required for the actin-dependent activities of Acanthamoeba myosin IC in vitro and indicate that the length of the side chain that delivers this charge is important. PMID- 9860947 TI - Structural invariance of constitutively active and inactive mutants of acanthamoeba myosin IC bound to F-actin in the rigor and ADP-bound states. AB - The three single-headed monomeric myosin I isozymes of Acanthamoeba castellanii (AMIs)-AMIA, AMIB, and AMIC-are among the best-studied of all myosins. We have used AMIC to study structural correlates of myosin's actin-activated ATPase. This activity is normally controlled by phosphorylation of Ser-329, but AMIC may be switched into constitutively active or inactive states by substituting this residue with Glu or Ala, respectively. To determine whether activation status is reflected in structural differences in the mode of attachment of myosin to actin, these mutant myosins were bound to actin filaments in the absence of nucleotide (rigor state) and visualized at 24-A resolution by using cryoelectron microscopy and image reconstruction. No such difference was observed. Consequently, we suggest that regulation may be affected not by altering the static (time averaged) structure of AMIC but by modulating its dynamic properties, i.e., molecular breathing. The tail domain of vertebrate intestinal brush-border myosin I has been observed to swing through 31 degrees on binding of ADP. However, it was predicted on grounds of differing kinetics that any such effects with AMIC should be small [Jontes, J. D., Ostap, E. M., Pollard, T. D. & Milligan, R. A. (1998) J. Cell Biol. 141, 155-162]. We have confirmed this hypothesis by observing actin-associated AMIC in its ADP-bound state. Finally, we compared AMIC to brush-border myosin I and AMIB, which were previously studied under similar conditions. In each case, the shape and angle of attachment to F-actin of the catalytic domain is largely conserved, but the domain structure and disposition of the tail is distinctively different for each myosin. PMID- 9860948 TI - Protein component of the ribozyme ribonuclease P alters substrate recognition by directly contacting precursor tRNA. AB - The protein component of ribonuclease P (RNase P) binds to the RNA subunit, forming a functional ribonucleoprotein complex in vivo and enhancing the affinity of the precursor tRNA (pre-tRNA) substrate. Photocrosslinking experiments with pre-tRNA bound to RNase P reconstituted with the protein component of Bacillus subtilis ribonuclease P (P protein) site specifically modified with a crosslinking reagent indicate that: (i) the central cleft of P protein directly interacts with the single-stranded 5' leader sequence of pre-tRNA, and (ii) the orientation and register of the pre-tRNA leader sequence in the central cleft places the protein component in close proximity to the active site. This unique mode of interaction suggests that the catalytic active site in RNase P occurs near the interface of RNA and protein. In contrast to other ribonucleoprotein complexes where the protein mainly stabilizes the active tertiary fold of the RNA, a critical function of the protein component of RNase P is to alter substrate specificity and enhance catalytic efficiency. PMID- 9860949 TI - Temperature, template topology, and factor requirements of archaeal transcription. AB - Although Archaea are prokaryotic and resemble Bacteria morphologically, their transcription apparatus is remarkably similar to those of eukaryotic cell nuclei. Because some Archaea exist in environments with temperatures of around 100 degreesC, they are likely to have evolved unique strategies for transcriptional control. Here, we investigate the effects of temperature and DNA template topology in a thermophilic archaeal transcription system. Significantly, and in marked contrast with characterized eucaryal systems, archaeal DNA template topology has negligible effect on transcription levels at physiological temperatures using highly purified polymerase and recombinant transcription factors. Furthermore, archaeal transcription does not require hydrolysis of the beta-gamma phosphoanhydride bond of ATP. However, at lower temperatures, negatively supercoiled templates are transcribed more highly than those that are positively supercoiled. Notably, the block to transcription on positively supercoiled templates at lowered temperatures is at the level of polymerase binding and promoter opening. These data imply that Archaea do not possess a functional homologue of transcription factor TFIIH, and that for the promoters studied, transcription is mediated by TATA box-binding protein, transcription factor TFB, and RNA polymerase alone. Furthermore, they suggest that the reduction of plasmid linking number by hyperthermophilic Archaea in vivo in response to cold shock is a mechanism to maintain gene expression under these adverse circumstances. PMID- 9860950 TI - Interaction of the Hsp70 molecular chaperone, DnaK, with its cochaperone DnaJ. AB - Chaperones of the Hsp70 family bind to unfolded or partially folded polypeptides to facilitate many cellular processes. ATP hydrolysis and substrate binding, the two key molecular activities of this chaperone, are modulated by the cochaperone DnaJ. By using both genetic and biochemical approaches, we provide evidence that DnaJ binds to at least two sites on the Escherichia coli Hsp70 family member DnaK: under the ATPase domain in a cleft between its two subdomains and at or near the pocket of substrate binding. The lower cleft of the ATPase domain is defined as a binding pocket for the J-domain because (i) a DnaK mutation located in this cleft (R167H) is an allele-specific suppressor of the binding defect of the DnaJ mutation, D35N and (ii) alanine substitution of two residues close to R167 in the crystal structure, N170A and T173A, significantly decrease DnaJ binding. A second binding determinant is likely to be in the substrate-binding domain because some DnaK mutations in the vicinity of the substrate-binding pocket are defective in either the affinity (G400D, G539D) or rate (D526N) of both peptide and DnaJ binding to DnaK. Binding of DnaJ may propagate conformational changes to the nearby ATPase catalytic center and substrate binding sites as well as facilitate communication between these two domains to alter the molecular properties of Hsp70. PMID- 9860951 TI - Mutations in the DnaK chaperone affecting interaction with the DnaJ cochaperone. AB - Hsp70 chaperones assist protein folding by ATP-controlled cycles of substrate binding and release. ATP hydrolysis is the rate-limiting step of the ATPase cycle that causes locking in of substrates into the substrate-binding cavity of Hsp70. This key step is strongly stimulated by DnaJ cochaperones. We show for the Escherichia coli Hsp70 homolog, DnaK, that stimulation by DnaJ requires the linked ATPase and substrate-binding domains of DnaK. Functional interaction with DnaJ is affected by mutations in an exposed channel located in the ATPase domain of DnaK. It is proposed that binding to this channel, possibly involving the J domain, allows DnaJ to couple substrate binding with ATP hydrolysis by DnaK. Evolutionary conservation of the channel and the J-domain suggests conservation of the mechanism of action of DnaJ proteins. PMID- 9860952 TI - Novel role of phosphorylation in Fe-S cluster stability revealed by phosphomimetic mutations at Ser-138 of iron regulatory protein 1. AB - Animals regulate iron metabolism largely through the action of the iron regulatory proteins (IRPs). IRPs modulate mRNA utilization by binding to iron responsive elements (IRE) in the 5' or 3' untranslated region of mRNAs encoding proteins involved in iron homeostasis or energy production. IRP1 is also the cytosolic isoform of aconitase. The activities of IRP1 are mutually exclusive and are modulated through the assembly/disassembly of its [4Fe-4S] cluster, reversibly converting it between an IRE-binding protein and cytosolic aconitase. IRP1 is also phosphoregulated by protein kinase C, but the mechanism by which phosphorylation posttranslationally increases IRE binding activity has not been fully defined. To investigate this, Ser-138 (S138), a PKC phosphorylation site, was mutated to phosphomimetic glutamate (S138E), aspartate (S138D), or nonphosphorylatable alanine (S138A). The S138E IRP1 mutant and, to a lesser extent, the S138D IRP1 mutant were impaired in aconitase function in yeast when grown aerobically but not when grown anaerobically. Purified wild-type and mutant IRP1s could be reconstituted to active aconitases anaerobically. However, when exposed to oxygen, the [4Fe-4S] cluster of the S138D and S138E mutants decayed 5 fold and 20-fold faster, respectively, than was observed for wild-type IRP1. Our findings suggest that stability of the Fe-S cluster of IRP1 can be regulated by phosphorylation and reveal a mechanism whereby the balance between the IRE binding and [4Fe-4S] forms of IRP1 can be modulated independently of cellular iron status. Furthermore, our results show that IRP1 can function as an oxygen modulated posttranscriptional regulator of gene expression. PMID- 9860953 TI - Bovine papillomavirus E5 protein induces oligomerization and trans phosphorylation of the platelet-derived growth factor beta receptor. AB - The bovine papillomavirus E5 protein is a 44-aa transmembrane protein that forms a stable complex with the cellular platelet-derived growth factor (PDGF) beta receptor and induces constitutive tyrosine phosphorylation and activation of the receptor, resulting in cell transformation. The E5 protein does not resemble PDGF, but rather activates the receptor in a ligand-independent fashion, thus providing a unique system to examine activation of receptor tyrosine kinases. Here, we used a variety of approaches to explore the mechanism of receptor activation by the E5 protein. Chemical cross-linking experiments revealed that the E5 protein activated only a small fraction of the endogenous PDGF beta receptor in transformed fibroblasts and suggested that this fraction was constitutively dimerized. Coimmunoprecipitation experiments using extracts of cells engineered to coexpress full-length and truncated PDGF beta receptors confirmed that the E5 protein induced oligomerization of the receptor. Furthermore, in cells expressing the E5 protein, a kinase-active receptor was able to trans-phosphorylate a kinase-negative mutant receptor but was unable to catalyze intramolecular autophosphorylation. These results indicated that the E5 protein induced PDGF beta receptor activation by forming a stable complex with the receptor, resulting in receptor dimerization and trans-phosphorylation. PMID- 9860954 TI - Stimulation of bacteriophage T4 middle transcription by the T4 proteins MotA and AsiA occurs at two distinct steps in the transcription cycle. AB - The bacteriophage T4 encodes proteins that are responsible for tightly regulating mRNA synthesis throughout phage development in Escherichia coli. The three classes of T4 promoters (early, middle, and late) are utilized sequentially by the host RNA polymerase as a result of phage-induced modifications. One such modification is the tight binding of the T4 AsiA protein to the sigma70 subunit of the RNA polymerase. This interaction is pivotal for the transition between T4 early and middle transcription, since it both inhibits recognition of host and T4 early promoters and stimulates T4 middle mode synthesis. The activation of T4 middle transcription also requires the T4 MotA protein, bound specifically to its recognition sequence, the "Mot box," which is centered at position -30 of these promoters. Accordingly, the two T4 proteins working in concert are sufficient to effectively switch the transcription specificity of the RNA polymerase holoenzyme. Herein, we investigate the mechanism of transcription activation and report that, while the presence of MotA and AsiA increases the initial recruitment of RNA polymerase to a T4 middle promoter, it does not alter the intrinsic stability of the discrete complexes formed. In addition, we have characterized the RNA polymerase-promoter species by UV laser footprinting and followed their evolution from open into initiating complexes. These data, combined with in vitro transcription assays, indicate that AsiA and MotA facilitate promoter escape, thereby stimulating the production of full-length transcripts. PMID- 9860955 TI - The biochemical properties of the ATPase activity of a 70-kDa heat shock protein (Hsp70) are governed by the C-terminal domains. AB - The cytosolic 70-kDa heat shock proteins (Hsp70s), Ssa and Ssb, of Saccharomyces cerevisiae are functionally distinct. Here we report that the ATPase activities of these two classes of Hsp70s exhibit different kinetic properties. The Ssa ATPase has properties similar to those of other Hsp70s studied, such as DnaK and Hsc70. Ssb, however, has an unusually low steady-state affinity for ATP but a higher maximal velocity. In addition, the ATPase activity of Hsp70s, like that of Ssa1, depends on the addition of K+ whereas Ssb activity does not. Suprisingly, the isolated 44-kDa ATPase domain of Ssb has a Km and Vmax for ATP hydrolysis similar to those of Ssa, rather than those of full length Ssb. Analysis of Ssa/Ssb fusion proteins demonstrates that the Ssb peptide-binding domain fused to the Ssa ATPase domain generates an ATPase of relatively high activity and low steady-state affinity for ATP similar to that of native Ssb. Therefore, at least some of the biochemical differences between the ATPases of these two classes of Hsp70s are not intrinsic to the ATPase domain itself. The differential influence of the peptide-binding domain on the ATPase domain may, in part, explain the functional uniqueness of these two classes of Hsp70s. PMID- 9860956 TI - Formation of the preprimosome protects lambda O from RNA transcription-dependent proteolysis by ClpP/ClpX. AB - Using the bacteriophage lambda DNA replication system, composed entirely of purified proteins, we have tested the accessibility of the short-lived lambda O protein to the ClpP/ClpX protease during the various stages of lambda DNA replication. We find that binding of lambda O protein to its orilambda DNA sequence, leading to the so-called "O-some" formation, largely inhibits its degradation. On the contrary, under conditions permissive for transcription, the lambda O protein bound to the orilambda sequence becomes largely accessible to ClpP/ClpX-mediated proteolysis. However, when the lambda O protein is part of the larger orilambda:O.P.DnaB preprimosomal complex, transcription does not significantly increase ClpP/ClpX-dependent lambda O degradation. These results show that transcription can stimulate proteolysis of a protein that is required for the initiation of DNA replication. PMID- 9860957 TI - Structural requirements for peptidic antagonists of the corticotropin-releasing factor receptor (CRFR): development of CRFR2beta-selective antisauvagine-30. AB - Different truncated and conformationally constrained analogs of corticotropin releasing factor (CRF) were synthesized on the basis of the amino acid sequences of human/rat CRF (h/rCRF), ovine CRF (oCRF), rat urocortin (rUcn), or sauvagine (Svg) and tested for their ability to displace [125I-Tyr0]oCRF or [125I-Tyr0]Svg from membrane homogenates of human embryonic kidney (HEK) 293 cells stably transfected with cDNA coding for rat CRF receptor, type 1 (rCRFR1), or mouse CRF receptor, type 2beta (mCRFR2beta). Furthermore, the potency of CRF antagonists to inhibit oCRF- or Svg-stimulated cAMP production of transfected HEK 293 cells expressing either rCRFR1 (HEK-rCRFR1 cells) or mCRFR2beta (HEK-mCRFR2beta cells) was determined. In comparison with astressin, which exhibited a similar affinity to rCRFR1 (Kd = 5.7 +/- 1.6 nM) and mCRFR2beta (Kd = 4.0 +/- 2.3 nM), [DPhe11,His12]Svg(11-40), [DLeu11]Svg(11-40), [DPhe11]Svg(11-40), and Svg(11-40) bound, respectively, with a 110-, 80-, 68-, and 54-fold higher affinity to mCRFR2beta than to rCRFR1. The truncated analogs of rUcn displayed modest preference (2- to 7-fold) for binding to mCRFR2beta. In agreement with the results of these binding experiments, [DPhe11, His12]Svg(11-40), named antisauvagine-30, was the most potent and selective ligand to suppress agonist induced adenylate cyclase activity in HEK cells expressing mCRFR2beta. PMID- 9860958 TI - The barrier-to-autointegration protein is a host factor for HIV type 1 integration. AB - In vivo, retroviral integration is mediated by a large nucleoprotein complex, termed the preintegration complex (PIC). PICs isolated from infected cells display in vitro integration activity. Here, we analyze the roles of different host cell factors in the structure and function of HIV type 1 (HIV-1) PICs. PICs purified by size exclusion after treatment with high salt lost their integration activity, and adding back an extract from uninfected cells restored this activity. In parallel, the native protein-DNA intasome structure detected at the ends of HIV-1 by Mu-mediated PCR footprinting was abolished by high salt and restored by the crude cell extract. Various purified proteins previously implicated in retroviral PIC function then were analyzed for their effects on the structure and function of salt-treated HIV-1 PICs. Whereas relatively low amounts (5-20 nM) of human barrier-to-autointegration factor (BAF) protein restored integration activity, substantially more (5-10 microM) human host factor HMG I(Y) was required. Similarly high levels (3-8 microM) of bovine RNase A, a DNA-binding protein used as a nonspecific control, also restored activity. Mu-mediated PCR footprinting revealed that of these three purified proteins, only BAF restored the native structure of the HIV-1 protein-DNA intasome. We suggest that BAF is a natural host cofactor for HIV-1 integration. PMID- 9860959 TI - Minimal and optimal mechanisms for GroE-mediated protein folding. AB - We have analyzed the effects of different components of the GroE chaperonin system on protein folding by using a nonpermissive substrate (i.e., one that has very low spontaneous refolding yield) for which rate data can be acquired. In the absence of GroES and nucleotides, the rate of GroEL-mediated refolding of heat- and DTT-denatured mitochondrial malate dehydrogenase was extremely low, but some three times higher than the spontaneous rate. This GroEL-mediated rate was increased 17-fold by saturating concentrations of ATP, 11-fold by ADP and GroES, and 465-fold by ATP and GroES. Optimal refolding activity was observed when the dissociation of GroES from the chaperonin complex was dramatically reduced. Although GroEL minichaperones were able to bind denatured mitochondrial malate dehydrogenase, they were ineffective in enhancing the refolding rate. The spectrum of mechanisms for GroE-mediated protein folding depends on the nature of the substrate. The minimal mechanism for permissive substrates (i.e., having significant yields of spontaneous refolding), requires only binding to the apical domain of GroEL. Slow folding rates of nonpermissive substrates are limited by the transitions between high- and low-affinity states of GroEL alone. The optimal mechanism, which requires holoGroEL, physiological amounts of GroES, and ATP hydrolysis, is necessary for the chaperonin-mediated folding of nonpermissive substrates at physiologically relevant rates under conditions in which retention of bound GroES prevents the premature release of aggregation-prone folding intermediates from the chaperonin complex. The different mechanisms are described in terms of the structural features of mini- and holo-chaperones. PMID- 9860960 TI - RNA polymerase subunit RPB5 plays a role in transcriptional activation. AB - A mutation in RPB5 (rpb5-9), an essential RNA polymerase subunit assembled into RNA polymerases I, II, and III, revealed a role for this subunit in transcriptional activation. Activation by GAL4-VP16 was impaired upon in vitro transcription with mutant whole-cell extracts. In vivo experiments using inducible reporter plasmids and Northern analysis support the in vitro data and demonstrate that RPB5 influences activation at some, but not all, promoters. Remarkably, this mutation maps to a conserved region of human RPB5 implicated by others to play a role in activation. Chimeric human-yeast RPB5 containing this conserved region now can function in place of its yeast counterpart. The defects noted with rpb5-9 are similar to those seen in truncation mutants of the RPB1 carboxyl terminal domain (CTD). We demonstrate that RPB5 and the RPB1-CTD have overlapping roles in activation because the double mutant is synthetically lethal and has exacerbated activation defects at the GAL1/10 promoter. These studies demonstrate that there are multiple activation targets in RNA polymerase II and that RPB5 and the CTD have similar roles in activation. PMID- 9860961 TI - Cloning and functional expression of a cDNA encoding a pheromone gland-specific acyl-CoA Delta11-desaturase of the cabbage looper moth, Trichoplusia ni. AB - Desaturation of coenzyme-A esters of saturated fatty acids is a common feature of sex pheromone biosynthetic pathways in the Lepidoptera. The enzymes that catalyze this step share several biochemical properties with the ubiquitous acyl-CoA Delta9-desaturases of animals and fungi, suggesting a common ancestral origin. Unlike metabolic acyl-CoA Delta9-desaturases, pheromone desaturases have evolved unusual regio- and stereoselective activities that contribute to the remarkable diversity of chemical structures used as pheromones in this large taxonomic group. In this report, we describe the isolation of a cDNA encoding a pheromone gland desaturase from the cabbage looper moth, Trichoplusia ni, a species in which all unsaturated pheromone products are produced via a Delta11Z-desaturation mechanism. The largest ORF of the approximately 1,250-bp cDNA encodes a 349-aa apoprotein (PDesat-Tn Delta11Z) with a predicted molecular mass of 40,240 Da. Its hydrophobicity profile is similar overall to those of rat and yeast Delta9 desaturases, suggesting conserved transmembrane topology. A 182-aa core domain delimited by conserved histidine-rich motifs implicated in iron-binding and catalysis has 72 and 58% similarity (including conservative substitutions) to acyl-CoA Delta9Z-desaturases of rat and yeast, respectively. Northern blot analysis revealed an approximately 1,250-nt PDesat-Tn Delta11Z mRNA that is consistent with the spatial and temporal distribution of Delta11-desaturase enzyme activity. Genetic transformation of a desaturase-deficient strain of the yeast Saccharomyces cerevisiae with an expression plasmid encoding PDesat-Tn Delta11Z resulted in complementation of the strain's fatty acid auxotrophy and the production of Delta11Z-unsaturated fatty acids. PMID- 9860963 TI - High-resolution NMR of encapsulated proteins dissolved in low-viscosity fluids. AB - The majority of known proteins are too large to be comprehensively examined by solution NMR methods, primarily because they tumble too slowly in solution. Here we introduce an approach to making the NMR relaxation properties of large proteins amenable to modern solution NMR techniques. The encapsulation of a protein in a reverse micelle dissolved in a low-viscosity fluid allows it to tumble as fast as a much smaller protein. The approach is demonstrated and validated with the protein ubiquitin encapsulated in reverse micelles prepared in a variety of alkane solvents. PMID- 9860962 TI - HNS, a nuclear-cytoplasmic shuttling sequence in HuR. AB - Proteins are transported into and out of the cell nucleus via specific signals. The two best-studied nuclear transport processes are mediated either by classical nuclear localization signals or nuclear export signals. There also are shuttling sequences that direct the bidirectional transport of RNA-binding proteins. Two examples are the M9 sequence in heterogeneous nuclear ribonucleoprotein A1 and the heterogeneous nuclear ribonucleoprotein K shuttling domain (KNS) sequence in heterogeneous nuclear ribonucleoprotein K, both of which appear to contribute importantly to the export of mRNA to the cytoplasm. HuR is an RNA-binding protein that can stabilize labile mRNAs containing AU-rich elements in their 3' untranslated regions and has been shown to shuttle between the nucleus and cytoplasm (18, 19). We have identified in HuR a shuttling sequence that also possess transcription-dependent nuclear localization signal activity. We propose that HuR first may bind AU-rich element-containing mRNAs in the nucleus and then escort them through the nuclear pore, providing protection during and after export to the cytoplasmic compartment. PMID- 9860964 TI - Gap junctional coupling in lenses lacking alpha3 connexin. AB - Fiber cells of the lens are interconnected by an extensive network of gap junctions containing alpha3 (Cx46) and alpha8 (Cx50) connexins. A specific role for these connexins in lens homeostasis is not known. To determine the contribution of these connexins to lens function, we used impedance techniques to study cell-to-cell coupling in lenses from homozygous alpha3 knockout (-/-), heterozygous (+/-), and wild-type (+/+) mice. Western blots and immunofluorescence data indicated that alpha8 remained at similar levels in the three classes of lenses, whereas alpha3 was approximately 50% of the normal level in the +/- lenses, and it was absent from the -/- lenses. Moreover, the data from +/+ lenses suggest that a cleavage of connexins occurs abruptly between the peripheral shell of differentiating fibers (DF) and the inner core of mature fibers (MF). The appearance of the cleaved connexins was correlated to a change in the coupling conductance. In -/- lenses the coupling conductance of MF was zero, and these fibers were depolarized by about 30 mV from normal (approximately -65 mV). The DF remained coupled, but the conductance was reduced to 30-35% of normal. However, the gap junctions in the DF of alpha3 -/- lenses remained sensitive to pH. We conclude that alpha3 connexin is necessary for the coupling of central fibers to peripheral cells, and that this coupling is essential for fiber cell homeostasis because uncoupled MF depolarize and subsequently become opaque. PMID- 9860965 TI - Conformational changes between the active-site and regulatory light chain of myosin as determined by luminescence resonance energy transfer: the effect of nucleotides and actin. AB - Myosin is thought to generate movement of actin filaments via a conformational change between its light-chain domain and its catalytic domain that is driven by the binding of nucleotides and actin. To monitor this change, we have measured distances between a gizzard regulatory light chain (Cys 108) and the active site (near or at Trp 130) of skeletal myosin subfragment 1 (S1) by using luminescence resonance energy transfer and a photoaffinity ATP-lanthanide analog. The technique allows relatively long distances to be measured, and the label enables site-specific attachment at the active-site with only modest affect on myosin's enzymology. The distance between these sites is 66.8 +/- 2.3 A when the nucleotide is ADP and is unchanged on binding to actin. The distance decreases slightly with ADP-BeF3, (-1.6 +/- 0.3 A) and more significantly with ADP-AlF4 ( 4.6 +/- 0.2 A). During steady-state hydrolysis of ATP, the distance is temperature-dependent, becoming shorter as temperature increases and the complex with ADP.Pi is favored over that with ATP. We conclude that the distance between the active site and the light chain varies as Acto-S1-ADP approximately S1-ADP > S1-ADP-BeF3 > S1-ADP-AlF4 approximately S1-ADP-Pi and that S1-ATP > S1-ADP-Pi. The changes in distance are consistent with a substantial rotation of the light chain binding domain of skeletal S1 between the prepowerstroke state, simulated by S1-ADP-AlF4, and the post-powerstroke state, simulated by acto-S1-ADP. PMID- 9860966 TI - Protein fluctuations are sensed by stimulated infrared echoes of the vibrations of carbon monoxide and azide probes. AB - The correlation functions of the fluctuations of vibrational frequencies of azide ions and carbon monoxide in proteins are determined directly from stimulated photon echoes generated with femtosecond infrared pulses. The asymmetric stretching vibration of azide bound to carbonic anhydrase II exhibits a pronounced evolution of its vibrational frequency distribution on the time scale of a few picoseconds, which is attributed to modifications of the ligand structure through interactions with the nearby Thr-199. When azide is bound in hemoglobin, a more complex evolution of the protein structure is required to interchange the different ligand configurations, as evidenced by the much slower relaxation of the frequency distribution in this case. The time evolution of the distribution of frequencies of carbon monoxide bound in hemoglobin occurs on the approximately 10-ps time scale and is very nonexponential. The correlation functions of the frequency fluctuations determine the evolution of the protein structure local to the probe and the extent to which the probe can navigate those parts of the energy landscape where the structural configurations are able to modify the local potential energy function of the probe. PMID- 9860967 TI - A model for amplification of hair-bundle motion by cyclical binding of Ca2+ to mechanoelectrical-transduction channels. AB - Amplification of auditory stimuli by hair cells augments the sensitivity of the vertebrate inner ear. Cell-body contractions of outer hair cells are thought to mediate amplification in the mammalian cochlea. In vertebrates that lack these cells, and perhaps in mammals as well, active movements of hair bundles may underlie amplification. We have evaluated a mathematical model in which amplification stems from the activity of mechanoelectrical-transduction channels. The intracellular binding of Ca2+ to channels is posited to promote their closure, which increases the tension in gating springs and exerts a negative force on the hair bundle. By enhancing bundle motion, this force partially compensates for viscous damping by cochlear fluids. Linear stability analysis of a six-state kinetic model reveals Hopf bifurcations for parameter values in the physiological range. These bifurcations signal conditions under which the system's behavior changes from a damped oscillatory response to spontaneous limit cycle oscillation. By varying the number of stereocilia in a bundle and the rate constant for Ca2+ binding, we calculate bifurcation frequencies spanning the observed range of auditory sensitivity for a representative receptor organ, the chicken's cochlea. Simulations using prebifurcation parameter values demonstrate frequency-selective amplification with a striking compressive nonlinearity. Because transduction channels occur universally in hair cells, this active channel model describes a mechanism of auditory amplification potentially applicable across species and hair-cell types. PMID- 9860968 TI - Cytologically normal cells from neoplastic cervical samples display extensive structural abnormalities on IR spectroscopy: implications for tumor biology. AB - Fourier-transform IR (FT-IR) spectra of pelleted exfoliated cervical cells from patients with cervical cancer or dysplasia differ from those from normal women. To study the origin of these spectral changes, we obtained the FT-IR spectra of individual cervical cells from normal, dysplastic, and malignant cervical samples. Ninety five percent of normal superficial and intermediate cells displayed two distinct spectral patterns designated A and B, and 5% displayed an intermediate pattern, suggesting extensive structural heterogeneity among these cells. Parabasal and endocervical cells showed pattern B spectra. The spectra of malignant, dysplastic, and other abnormal cells also were characterized. Analysis of FT-IR spectra of over 2, 000 individual cells from 10 normal females, 7 females with dysplasia, and 5 females with squamous cell carcinoma revealed that the spectra of normal-appearing intermediate and superficial cells of the cervix from women with either dysplasia or cancer differed from those of normal women. Chemometric and classical spectroscopic analysis showed a continuum of changes paralleling the transition from normalcy to malignancy. These findings suggest that (i) the structural changes underlying the spectroscopic changes are involved in or are a product of cervical carcinogenesis and (ii) the neoplastic process may be more extensive than currently recognized with morphological criteria. This approach may be useful for the structural study of neoplasia and also may be of help in the diagnosis or classification of cervical disorders. PMID- 9860969 TI - Plasticity of the neoplastic phenotype in vivo is regulated by epigenetic factors. AB - Age of host and transplantation-site microenvironment influence the tumorigenic potential of neoplastically transformed liver epithelial cells. Tumorigenic BAG2 GN6TF rat liver epithelial cells consistently form tumors at ectopic sites, but differentially express tumorigenicity or hepatocytic differentiation in the liver depending on host age and route of cell transplantation into the liver. Direct inoculation into host livers concentrates tumor cells locally, resulting in undifferentiated tumors near the transplantation site in both young (3-month-old) and old (18-month-old) rats. Transplantation-site tumors regress within 1 month in the livers of young rats, but grow progressively in old rats. However, inoculation of cells into the spleen distributes transplanted cells individually throughout the liver, resulting in hepatocytic differentiation by tumor cells with concomitant suppression of their tumorigenicity in young rats. When transplanted into livers of old rats by splenic inoculation, or when young hepatic-transplant recipients are allowed to age, hepatocytic progeny of BAG2 GN6TF cells proliferate to form foci, suggesting that the liver microenvironment of old rats incompletely regulates the proliferation and differentiation of tumor cell-derived hepatocytes. Upon removal from the liver, BAG2-GN6TF-derived hepatocytes revert to an undifferentiated, aggressively tumorigenic phenotype. We posit that the spectrum between normal differentiation and malignant potential of these cells reflects the dynamic interaction of the specific transformation related genotype of the cells and the characteristics of the tissue microenvironment at the transplantation site. Changes in the tissue milieu, such as those that accompany normal aging, may determine the ability of a genetically aberrant cell to produce a tumor. PMID- 9860970 TI - Inhibition of beta-catenin-mediated transactivation by cadherin derivatives. AB - We studied the effect of N-cadherin, and its free or membrane-anchored cytoplasmic domain, on the level and localization of beta-catenin and on its ability to induce lymphocyte enhancer-binding factor 1 (LEF-1)-responsive transactivation. These cadherin derivatives formed complexes with beta-catenin and protected it from degradation. N-cadherin directed beta-catenin into adherens junctions, and the chimeric protein induced diffuse distribution of beta-catenin along the membrane whereas the cytoplasmic domain of N-cadherin colocalized with beta-catenin in the nucleus. Cotransfection of beta-catenin and LEF-1 into Chinese hamster ovary cells induced transactivation of a LEF-1 reporter, which was blocked by the N-cadherin-derived molecules. Expression of N-cadherin and an interleukin 2 receptor/cadherin chimera in SW480 cells relocated beta-catenin from the nucleus to the plasma membrane and reduced transactivation. The cytoplasmic tails of N- or E-cadherin colocalized with beta-catenin in the nucleus, and suppressed the constitutive LEF-1-mediated transactivation, by blocking beta-catenin-LEF-1 interaction. Moreover, the 72 C-terminal amino acids of N-cadherin stabilized beta-catenin and reduced its transactivation potential. These results indicate that beta-catenin binding to the cadherin cytoplasmic tail either in the membrane, or in the nucleus, can inhibit beta-catenin degradation and efficiently block its transactivation capacity. PMID- 9860972 TI - Aldol sensors for the rapid generation of tunable fluorescence by antibody catalysis. AB - The synthesis of novel fluorogenic retro-aldol substrates for aldolase antibody 38C2 is described. These substrates are efficiently and specifically processed by antibody aldolases but not by natural cellular enzymes. Together, the fluorogenic substrates and antibody aldolases provide reporter gene systems that are compatible with living cells. The broad scope of the antibody aldolase allows for the processing of a range of substrates that can be designed to allow fluorescence monitoring at a variety of wavelengths. We also have developed the following concept in fluorescent protein tags. beta-Diketones bearing a fluorescent tag are bound covalently by the aldolase antibody and not other proteins. We anticipate that proteins fused with the antibody can be tagged specifically and covalently within living cells with fluorophores of virtually any color, thereby providing an alternative to green fluorescent protein fusions. PMID- 9860971 TI - Otoconin-90, the mammalian otoconial matrix protein, contains two domains of homology to secretory phospholipase A2. AB - The ability to sense orientation relative to gravity requires dense particles, called otoconia, which are localized in the vestibular macular organs. In mammals, otoconia are composed of proteins (otoconins) and calcium carbonate crystals in a calcite lattice. Little is known about the mechanisms that regulate otoconial biosynthesis. To begin to elucidate these mechanisms, we have partially sequenced and cloned the major protein component of murine otoconia, otoconin-90 (OC90). The amino acid sequence identified an orphan chimeric human cDNA. Because of its similarity to secretory phospholipase A2 (sPLA2), this gene was referred to as PLA2-like (PLA2L) and enabled the identification of human Oc90. Partial murine cDNA and genomic clones were isolated and shown to be specifically expressed in the developing mouse otocyst. The mature mouse OC90 is composed of 453 residues and contains two domains homologous to sPLA2. The cloning of Oc90 will allow an examination of the role of this protein in otoconial biosynthesis and in diseases that affect the vestibular system. PMID- 9860973 TI - Farnesyltransferase inhibitors induce cytochrome c release and caspase 3 activation preferentially in transformed cells. AB - Farnesyltransferase inhibitors (FTIs) represent a new class of anticancer drugs that show promise in blocking the growth of tumors. Here, we report that FTIs are capable of inducing apoptosis of transformed but not untransformed cells. Treatment of v-K-ras-transformed normal rat kidney (KNRK) cells with FTIs leads to the induction of apoptotic cell morphology, chromatin condensation and DNA fragmentation. In addition, fluorescence-activated cell sorter analysis of FTI treated KNRK cells shows a sub-G1 apoptotic peak (chromosome content of <2 N). This FTI-induced apoptosis is evident only when the cells are grown in low serum conditions (0.1% fetal calf serum) and is observed selectively with transformed KNRK cells and not with untransformed NRK cells. Further analysis of the mechanism underlying this apoptosis has shown that FTI treatment of KNRK cells results in the activation of caspase 3 but not caspase 1. Moreover, the addition of Z-DEVD-fmk, an agent that interferes with caspase 3 activity, can inhibit FTI induced apoptosis in a dose-dependent manner. Introduction of the CASP-3 gene into MCF7 cells, which lack caspase 3 activity, results in a significant increase of FTI-induced apoptosis. Furthermore, FTI induces the release of cytochrome c into the cytosol. This release is an important feature of caspase 3-mediated apoptosis. These results suggest that FTIs induce apoptosis through the release of cytochrome c from the mitochondria resulting in caspase 3 activation. PMID- 9860974 TI - The Arp2/3 complex mediates actin polymerization induced by the small GTP-binding protein Cdc42. AB - The small GTP-binding protein Cdc42 is thought to induce filopodium formation by regulating actin polymerization at the cell cortex. Although several Cdc42 binding proteins have been identified and some of them have been implicated in filopodium formation, the precise role of Cdc42 in modulating actin polymerization has not been defined. To understand the biochemical pathways that link Cdc42 to the actin cytoskeleton, we have reconstituted Cdc42-induced actin polymerization in Xenopus egg extracts. Using this cell-free system, we have developed a rapid and specific assay that has allowed us to fractionate the extract and isolate factors involved in this activity. We report here that at least two biochemically distinct components are required, based on their chromatographic behavior and affinity for Cdc42. One component is purified to homogeneity and is identified as the Arp2/3 complex, a protein complex that has been shown to nucleate actin polymerization. However, the purified complex alone is not sufficient to mediate the activity; a second component that binds Cdc42 directly and mediates the interaction between Cdc42 and the complex also is required. These results establish an important link between a signaling molecule, Cdc42, and a complex that can directly modulate actin networks in vitro. We propose that activation of the Arp2/3 complex by Cdc42 and other signaling molecules plays a central role in stimulating actin polymerization at the cell surface. PMID- 9860975 TI - Real-time quantitative measurement of autocrine ligand binding indicates that autocrine loops are spatially localized. AB - Autocrine ligands are important regulators of many normal tissues and have been implicated in a number of disease states, including cancer. However, because by definition autocrine ligands are synthesized, secreted, and bound to cell receptors within an intrinsically self-contained "loop," standard pharmacological approaches cannot be used to investigate relationships between ligand/receptor binding and consequent cellular responses. We demonstrate here a new approach for measurement of autocrine ligand binding to cells, using a microphysiometer assay originally developed for investigating cell responses to exogenous ligands. This technique permits quantitative measurements of autocrine responses on the time scale of receptor binding and internalization, thus allowing investigation of the role of receptor trafficking and dynamics in cellular responses. We used this technique to investigate autocrine signaling through the epidermal growth factor receptor by transforming growth factor alpha (TGFalpha) and found that anti receptor antibodies are far more effective than anti-ligand antibodies in inhibiting autocrine signaling. This result indicates that autocrine-based signals can operate in a spatially restricted, local manner and thus provide cells with information on their local microenvironment. PMID- 9860977 TI - A transgenic mouse model of metastatic prostate cancer originating from neuroendocrine cells. AB - A transgenic mouse model of metastatic prostate cancer has been developed that is 100% penetrant in multiple pedigrees. Nucleotides -6500 to +34 of the mouse cryptdin-2 gene were used to direct expression of simian virus 40 T antigen to a subset of neuroendocrine cells in all lobes of the FVB/N mouse prostate. Transgene expression is initiated between 7 and 8 weeks of age and leads to development of prostatic intraepithelial neoplasia within a week. Prostatic intraepithelial neoplasia progresses rapidly to local invasion. Metastases to lymph nodes, liver, lung, and bone are common by 6 months. Tumorigenesis is not dependent on androgens. This model indicates that the neuroendocrine cell lineage of the prostate is exquisitely sensitive to transformation and provides insights about the significance of neuroendocrine differentiation in human prostate cancer. PMID- 9860976 TI - The cyclin-dependent kinase inhibitor p27(Kip1) induces N-terminal proteolytic cleavage of cyclin A. AB - Progression through the cell cycle is regulated in part by the sequential activation and inactivation of cyclin-dependent kinases (CDKs). Many signals arrest the cell cycle through inhibition of CDKs by CDK inhibitors (CKIs). p27(Kip1) (p27) was first identified as a CKI that binds and inhibits cyclin A/CDK2 and cyclin E/CDK2 complexes in G1. Here we report that p27 has an additional property, the ability to induce a proteolytic activity that cleaves cyclin A, yielding a truncated cyclin A lacking the mitotic destruction box. Other CKIs (p15(Ink4b), p16(Ink4a), p21(Cip1), and p57(Kip2)) do not induce cleavage of cyclin A; other cyclins (cyclin B, D1, and E) are not cleaved by the p27-induced protease activity. The C-terminal half of p27, which is dispensable for its kinase inhibitory activity, is required to induce cleavage. Mechanistically, p27 does not appear to cause cleavage through direct interaction with cyclin/CDK complexes. Instead, it activates a latent protease that, once activated, does not require the continuing presence of p27. Mutation of cyclin A at R70 or R71, residues at or very close to the cleavage site, blocks cleavage. Noncleavable mutants are still recognized by the anaphase-promoting complex/cyclosome pathway responsible for ubiquitin-dependent proteolysis of mitotic cyclins, indicating that the p27-induced cleavage of cyclin A is part of a separate pathway. We refer to this protease as Tsap (pTwenty-seven- activated protease). PMID- 9860978 TI - A protein required for nuclear-protein import, Mog1p, directly interacts with GTP Gsp1p, the Saccharomyces cerevisiae ran homologue. AB - We previously isolated 25 temperature-sensitive gsp1 alleles of Saccharomyces cerevisiae Ran homologue, each of which possesses amino acid changes that differ from each other. We report here isolation of three multicopy suppressors-PDE2, NTF2, and a gene designated MOG1-all of which rescued a growth defect of these gsp1 strains. The gsp1 suppression occurred even in the absence of GSP2, another S. cerevisiae GSP1-like gene. Previously, NTF2 was reported to suppress gsp1 but not PDE2. Mog1p, with a calculated molecular mass of 24 kDa, was found to be encoded by the yeast ORF YJR074W. Both MOG1 and NTF2 suppressed a series of gsp1 alleles with similar efficiency, and both suppressed gsp1 even with a single gene dose. Consistent with the high efficiency of gsp1 suppression, Mog1p directly bound to GTP, but not to GDP-Gsp1p. The disruption of MOG1 made yeast temperature sensitive for growth. Deltamog1, which was suppressed by overexpression of NTF2, was found to have a defect in both classic and nonclassic nuclear localization signal-dependent nuclear-protein imports, but not in mRNA export. Thus, Mog1p, which was localized in the nucleus, is a Gsp1p-binding protein involved in nuclear-protein import and that functionally interacts with Ntf2p. Furthermore, the finding that PDE2 suppressed both gsp1 and rna1-1 indicates that the Ran GTPase cycle is regulated by the Ras-cAMP pathway. PMID- 9860979 TI - The adaptor protein Crk connects multiple cellular stimuli to the JNK signaling pathway. AB - c-Jun N-terminal kinases (JNKs) are potently activated by a number of cellular stimuli. Small GTPases, in particular Rac, are responsible for initiating the activation of the JNK pathways. So far, the signals leading from extracellular stimuli to the activation of Rac have remained elusive. Recent studies have demonstrated that the Src homology 2 (SH2)- and Src homology 3 (SH3)-containing adaptor protein Crk is capable of activating JNK when ectopically expressed. We found here that transient expression of Crk induces JNK activation, and this activation was dependent on both the SH2- and SH3-domains of Crk. Expression of p130(Cas) (Cas), a major binding protein for the Crk SH2-domain, also induced JNK activation, which was blocked by the SH2-mutant of Crk. JNK activation by Cas and Crk was effectively blocked by a dominant-negative form of Rac, suggesting for a linear pathway from the Cas-Crk-complex to the Rac-JNK activation. Many of the stimuli that activate the Rac-JNK pathway enhance engagement of the Crk SH2 domain. JNK activation by these stimuli, such as epidermal growth factor, integrin ligand binding and v-Src, was efficiently blocked by dominant-negative mutants of Crk. A dominant-negative form of Cas in turn blocked the integrin-, but not epidermal growth factor - nor v-Src-mediated JNK activation. Together, these results demonstrate an important role for Crk in connecting multiple cellular stimuli to the Rac-JNK pathway, and a role for the Cas-Crk complex in integrin-mediated JNK activation. PMID- 9860981 TI - PTEN/MMAC1/TEP1 suppresses the tumorigenicity and induces G1 cell cycle arrest in human glioblastoma cells. AB - PTEN/MMAC1/TEP1 is a tumor suppressor that possesses intrinsic phosphatase activity. Deletions or mutations of its encoding gene are associated with a variety of human cancers. However, very little is known about the molecular mechanisms by which this important tumor suppressor regulates cell growth. Here, we show that PTEN expression potently suppressed the growth and tumorigenicity of human glioblastoma U87MG cells. The growth suppression activity of PTEN was mediated by its ability to block cell cycle progression in the G1 phase. Such an arrest correlated with a significant increase of the cell cycle kinase inhibitor p27(KIP1) and a concomitant decrease in the activities of the G1 cyclin-dependent kinases. PTEN expression also led to the inhibition of Akt/protein kinase B, a serine-threonine kinase activated by the phosphatidylinositol 3-kinase (PI 3 kinase) signaling pathway. In addition, the effect of PTEN on p27(KIP1) and the cell cycle can be mimicked by treatment of U87MG cells with LY294002, a selective inhibitor of PI 3-kinase. Taken together, our studies suggest that the PTEN tumor suppressor modulates G1 cell cycle progression through negatively regulating the PI 3-kinase/Akt signaling pathway, and one critical target of this signaling process is the cyclin-dependent kinase inhibitor p27(KIP1). PMID- 9860980 TI - Differential regulation of transcription: repression by unactivated mitogen activated protein kinase Kss1 requires the Dig1 and Dig2 proteins. AB - Kss1, a yeast mitogen-activated protein kinase (MAPK), in its unphosphorylated (unactivated) state binds directly to and represses Ste12, a transcription factor necessary for expression of genes whose promoters contain filamentous response elements (FREs) and genes whose promoters contain pheromone response elements (PREs). Herein we show that two nuclear proteins, Dig1 and Dig2, are required cofactors in Kss1-imposed repression. Dig1 and Dig2 cooperate with Kss1 to repress Ste12 action at FREs and regulate invasive growth in a naturally invasive strain. Kss1-imposed Dig-dependent repression of Ste12 also occurs at PREs. However, maintenance of repression at PREs is more dependent on Dig1 and/or Dig2 and less dependent on Kss1 than repression at FREs. In addition, derepression at PREs is more dependent on MAPK-mediated phosphorylation than is derepression at FREs. Differential utilization of two types of MAPK-mediated regulation (binding imposed repression and phosphorylation-dependent activation), in combination with distinct Ste12-containing complexes, contributes to the mechanisms by which separate extracellular stimuli that use the same MAPK cascade can elicit two different transcriptional responses. PMID- 9860982 TI - A lineage-specific protein kinase crucial for myeloid maturation. AB - To identify genes involved in macrophage development, we used the differential display technique and compared the gene expression profiles for human myeloid HL 60 leukemia cell lines susceptible and resistant to macrophage maturation. We identified a gene coding for a protein kinase, protein kinase X (PRKX), which was expressed in the maturation-susceptible, but not in the resistant, cell line. The expression of the PRKX gene was found to be induced during monocyte, macrophage, and granulocyte maturation of HL-60 cells. We also studied the expression of the PRKX gene in 12 different human tissues and transformed cell lines and found that, among these tissues and cell types, the PRKX gene is expressed only in blood. Among the blood cell lineages, the PRKX gene is specifically expressed in macrophages and granulocytes. Antisense inhibition of PRKX expression blocked terminal development in both the leukemic HL-60 cells and normal peripheral blood monocytes, implying that PRKX is a key mediator of macrophage and granulocyte maturation. Using the HL-60 cell variant deficient in protein kinase C-beta (PKC beta) and several stable PKC-beta transfectants, we found that PRKX gene expression is under control of PKC-beta; hence PRKX is likely to act downstream of this PKC isozyme in the same signal transduction pathway leading to macrophage maturation. PMID- 9860983 TI - Mouse deformed epidermal autoregulatory factor 1 recruits a LIM domain factor, LMO-4, and CLIM coregulators. AB - Nuclear LIM domains interact with a family of coregulators referred to as Clim/Ldb/Nli. Although one family member, Clim-2/Ldb-1/Nli, is highly expressed in epidermal keratinocytes, no nuclear LIM domain factor is known to be expressed in epidermis. Therefore, we used the conserved LIM-interaction domain of Clim coregulators to screen for LIM domain factors in adult and embryonic mouse skin expression libraries and isolated a factor that is highly homologous to the previously described LIM-only proteins LMO-1, -2, and -3. This factor, referred to as LMO-4, is expressed in overlapping manner with Clim-2 in epidermis and in several other regions, including epithelial cells of the gastrointestinal, respiratory and genitourinary tracts, developing cartilage, pituitary gland, and discrete regions of the central and peripheral nervous system. Like LMO-2, LMO-4 interacts strongly with Clim factors via its LIM domain. Because LMO/Clim complexes are thought to regulate gene expression by associating with DNA-binding proteins, we used LMO-4 as a bait to screen for such DNA-binding proteins in epidermis and isolated the mouse homologue of Drosophila Deformed epidermal autoregulatory factor 1 (DEAF-1), a DNA-binding protein that interacts with regulatory sequences first described in the Deformed epidermal autoregulatory element. The interaction between LMO-4 and mouse DEAF-1 maps to a proline-rich C terminal domain of mouse DEAF-1, distinct from the helix-loop-helix and GATA domains previously shown to interact with LMOs, thus defining an additional LIM interacting domain. PMID- 9860986 TI - Natal origins of migratory monarch butterflies at wintering colonies in Mexico: new isotopic evidence. AB - Each year, millions of monarch butterflies from eastern North America migrate to overwinter in 10-13 discrete colonies located in the Oyamel forests of central Mexico. For decades efforts to track monarch migration have relied on observations and tag-recapture methods, culminating with the discovery of the wintering colonies in 1975. Monarch tag returns from Mexico, however, are few and primarily from two accessible colonies, and therefore tag-recapture techniques have not quantified natal origins or distinctiveness among monarch populations at wintering sites. Such information would be invaluable in the conservation of the monarch and its migration phenomenon since the wintering sites currently are threatened by habitat alteration. Here we show that stable hydrogen (deltaD) and carbon (delta13C) isotope ratios of wintering monarchs can be used to evaluate natal origins on the summer breeding range. Stable-hydrogen and carbon isotopic values of 597 wintering monarchs from 13 wintering roost sites were compared with isotopic patterns measured in individuals at natal sites across their breeding range over a single migration cycle. We determined that all monarch wintering colonies were composed of individuals originating mainly from the Midwest, United States, thereby providing evidence for a panmictic model of wintering colony composition. However, two colonies showed more northerly origins, suggesting possible priority colonies for conservation efforts. PMID- 9860985 TI - From patterns to processes: phase and density dependencies in the Canadian lynx cycle. AB - Across the boreal forest of North America, lynx populations undergo 10-year cycles. Analysis of 21 time series from 1821 to the present demonstrates that these fluctuations are generated by nonlinear processes with regulatory delays. Trophic interactions between lynx and hares cause delayed density-dependent regulation of lynx population growth. The nonlinearity, in contrast, appears to arise from phase dependencies in hunting success by lynx through the cycle. Using a combined approach of empirical, statistical, and mathematical modeling, we highlight how shifts in trophic interactions between the lynx and the hare generate the nonlinear process primarily by shifting functional response curves during the increase and the decrease phases. PMID- 9860984 TI - Metabolism to a response pathway selective retinoid ligand during axial pattern formation. AB - We report identification of 9-cis-4-oxo-retinoic acid (9-cis-4-oxo-RA) as an in vivo retinoid metabolite in Xenopus embryos. 9-Cis-4-oxo-RA bound receptors (RARs) alpha, beta, and gamma as well as retinoid X receptors (RXRs) alpha, beta, and gamma in vitro. However, this retinoid displayed differential RXR activation depending on the response pathway used. Although it failed to activate RXRs in RXR homodimers, it activated RXRs and RARs synergistically in RAR-RXR heterodimers. 9-Cis-4-oxo-RA thus acted as a dimer-specific agonist. Considering that RAR-RXR heterodimers are major functional units involved in transducing retinoid signals during embryogenesis and that 9-cis-4-oxo-RA displayed high potency for modulating axial pattern formation in Xenopus, metabolism to 9-cis-4 oxo-RA may provide a mechanism to target retinoid action to this and other RAR RXR heterodimer-mediated processes. PMID- 9860987 TI - Molecular evidence for an African origin of the Hawaiian endemic Hesperomannia (Asteraceae). AB - Identification of the progenitors of plants endemic to oceanic islands often is complicated by extreme morphological divergence between island and continental taxa. This is especially true for the Hawaiian Islands, which are 3,900 km from any continental source. We examine the origin of Hesperomannia, a genus of three species endemic to Hawaii that always have been placed in the tribe Mutisieae of the sunflower family. Phylogenetic analyses of representatives from all tribes in this family using the chloroplast gene ndhF (where ndhF is the ND5 protein of chloroplast NADH dehydrogenase) indicate that Hesperomannia belongs to the tribe Vernonieae. Phylogenetic comparisons within the Vernonieae using sequences of both ndhF and the internal transcribed spacer regions of nuclear ribosomal DNA reveal that Hesperomannia is sister to African species of Vernonia. Long-distance dispersal northeastward from Africa to southeast Asia and across the many Pacific Ocean island chains is the most likely explanation for this unusual biogeographic connection. The 17- to 26-million-year divergence time between African Vernonia and Hesperomannia estimated by the DNA sequences predates the age of the eight existing Hawaiian Islands. These estimates are consistent with an hypothesis that the progenitor of Hesperomannia arrived at one of the low islands of the Hawaiian Emperor chain between the late Oligocene and mid-Miocene when these islands were above sea level. Subsequent to its arrival the southeast Pacific island chains served as steppingstones for dispersal to the existing Hawaiian Islands. PMID- 9860988 TI - Comparative studies on mammalian Hoxc8 early enhancer sequence reveal a baleen whale-specific deletion of a cis-acting element. AB - Variations in regulatory regions of developmental control genes have been implicated in the divergence of axial morphologies. To find potentially significant changes in cis-regulatory regions, we compared nucleotide sequences and activities of mammalian Hoxc8 early enhancers. The nucleotide sequence of the early enhancer region is extremely conserved among mammalian clades, with five previously described cis-acting elements, A-E, being invariant. However, a 4-bp deletion within element C of the Hoxc8 early enhancer sequence is observed in baleen whales. When assayed in transgenic mouse embryos, a baleen whale enhancer (unlike other mammalian enhancers) directs expression of the reporter gene to more posterior regions of the neural tube but fails to direct expression to posterior mesoderm. We suggest that regulation of Hoxc8 in baleen whales differs from other mammalian species and may be associated with variation in axial morphology. PMID- 9860989 TI - Using rare mutations to estimate population divergence times: a maximum likelihood approach. AB - In this paper we propose a method to estimate by maximum likelihood the divergence time between two populations, specifically designed for the analysis of nonrecurrent rare mutations. Given the rapidly growing amount of data, rare disease mutations affecting humans seem the most suitable candidates for this method. The estimator RD, and its conditional version RDc, were derived, assuming that the population dynamics of rare alleles can be described by using a birth death process approximation and that each mutation arose before the split of a common ancestral population into the two diverging populations. The RD estimator seems more suitable for large sample sizes and few alleles, whose age can be approximated, whereas the RDc estimator appears preferable when this is not the case. When applied to three cystic fibrosis mutations, the estimator RD could not exclude a very recent time of divergence among three Mediterranean populations. On the other hand, the divergence time between these populations and the Danish population was estimated to be, on the average, 4,500 or 15,000 years, assuming or not a selective advantage for cystic fibrosis carriers, respectively. Confidence intervals are large, however, and can probably be reduced only by analyzing more alleles or loci. PMID- 9860990 TI - Evaluating multiple alternative hypotheses for the origin of Bilateria: an analysis of 18S rRNA molecular evidence. AB - Six alternative hypotheses for the phylogenetic origin of Bilateria are evaluated by using complete 18S rRNA gene sequences for 52 taxa. These data suggest that there is little support for three of these hypotheses. Bilateria is not likely to be the sister group of Radiata or Ctenophora, nor is it likely that Bilateria gave rise to Cnidaria or Ctenophora. Instead, these data reveal a close relationship between bilaterians, placozoans, and cnidarians. From this, several inferences can be drawn. Morphological features that previously have been identified as synapomorphies of Bilateria and Ctenophora, e.g., mesoderm, more likely evolved independently in each clade. The endomesodermal muscles of bilaterians may be homologous to the endodermal muscles of cnidarians, implying that the original bilaterian mesodermal muscles were myoepithelial. Placozoans should have a gastrulation stage during development. Of the three hypotheses that cannot be falsified with the 18S rRNA data, one is most strongly supported. This hypothesis states that Bilateria and Placozoa share a more recent common ancestor than either does to Cnidaria. If true, the simplicity of placozoan body architecture is secondarily derived from a more complex ancestor. This simplification may have occurred in association with a planula-type larva becoming reproductive before metamorphosis. If this simplification took place during the common history that placozoans share with bilaterians, then placozoan genes that contain a homeobox, such as Trox2, should be explored, for they may include the gene or genes most closely related to Hox genes of bilaterians. PMID- 9860991 TI - A new Eocene archaeocete (Mammalia, Cetacea) from India and the time of origin of whales. AB - Himalayacetus subathuensis is a new pakicetid archaeocete from the Subathu Formation of northern India. The type dentary has a small mandibular canal indicating a lack of auditory specializations seen in more advanced cetaceans, and it has Pakicetus-like molar teeth suggesting that it fed on fish. Himalayacetus is significant because it is the oldest archaeocete known and because it was found in marine strata associated with a marine fauna. Himalayacetus extends the fossil record of whales about 3.5 million years back in geological time, to the middle part of the early Eocene [ approximately 53.5 million years ago (Ma)]. Oxygen in the tooth-enamel phosphate has an isotopic composition intermediate between values reported for freshwater and marine archaeocetes, indicating that Himalayacetus probably spent some time in both environments. When the temporal range of Archaeoceti is calibrated radiometrically, comparison of likelihoods constrains the time of origin of Archaeoceti and hence Cetacea to about 54-55 Ma (beginning of the Eocene), whereas their divergence from extant Artiodactyla may have been as early as 64-65 Ma (beginning of the Cenozoic). PMID- 9860992 TI - Biosynthesis of the Pseudomonas polyketide coronafacic acid requires monofunctional and multifunctional polyketide synthase proteins. AB - Coronafacic acid (CFA) is the polyketide component of the phytotoxin coronatine, a virulence factor of the plant pathogen Pseudomonas syringae. Our current knowledge of polyketide biosynthesis largely is based on the analysis of polyketide synthases (PKSs) in actinomycetes and other Gram-positive bacteria. Consequently, the cloning and characterization of the CFA biosynthetic gene cluster will contribute significantly to our knowledge of polyketide synthesis in Pseudomonas. In this report, we describe two genes in the CFA biosynthetic gene cluster that encode PKSs that are structurally and functionally similar to the multifunctional modular PKSs, which catalyze the synthesis of macrolide antibiotics. The CFA PKS genes were overproduced in Escherichia coli and shown to cross-react with antisera made to a modular PKS involved in erythromycin synthesis. A scheme for CFA biosynthesis is presented that incorporates the activities of all proteins in the CFA PKS. In this report a gene cluster encoding a pseudomonad polyketide has been completely sequenced and the deduced gene functions have been used to develop a biosynthetic scheme. PMID- 9860993 TI - Bidirectional imprinting of a single gene: GNAS1 encodes maternally, paternally, and biallelically derived proteins. AB - The GNAS1 gene encodes the alpha subunit of the guanine nucleotide-binding protein Gs, which couples signaling through peptide hormone receptors to cAMP generation. GNAS1 mutations underlie the hormone resistance syndrome pseudohypoparathyroidism type Ia (PHP-Ia), so the maternal inheritance displayed by PHP-Ia has raised suspicions that GNAS1 is imprinted. Despite this suggestion, in most tissues Gsalpha is biallelically encoded. In contrast, the large G protein XLalphas, also encoded by GNAS1, is paternally derived. Because the inheritance of PHP-Ia predicts the existence of maternally, rather than paternally, expressed transcripts, we have investigated the allelic origin of other mRNAs derived from GNAS1. We find this gene to be remarkable in the complexity of its allele-specific regulation. Two upstream promoters, each associated with a large coding exon, lie only 11 kb apart, yet show opposite patterns of allele-specific methylation and monoallelic transcription. The more 5' of these exons encodes the neuroendocrine secretory protein NESP55, which is expressed exclusively from the maternal allele. The NESP55 exon is 11 kb 5' to the paternally expressed XLalphas exon. The transcripts from these two promoters both splice onto GNAS1 exon 2, yet share no coding sequences. Despite their structural unrelatedness, the encoded proteins, of opposite allelic origin, both have been implicated in regulated secretion in neuroendocrine tissues. Remarkably, maternally (NESP55), paternally (XLalphas), and biallelically (Gsalpha) derived proteins all are produced by different patterns of promoter use and alternative splicing of GNAS1, a gene showing simultaneous imprinting in both the paternal and maternal directions. PMID- 9860994 TI - rexB of bacteriophage lambda is an anti-cell death gene. AB - In Escherichia coli, programmed cell death is mediated through "addiction modules" consisting of two genes; the product of one gene is long-lived and toxic, whereas the product of the other is short-lived and antagonizes the toxic effect. Here we show that the product of lambdarexB, one of the few genes expressed in the lysogenic state of bacteriophage lambda, prevents cell death directed by each of two addiction modules, phd-doc of plasmid prophage P1 and the rel mazEF of E. coli, which is induced by the signal molecule guanosine 3',5' bispyrophosphate (ppGpp) and thus by amino acid starvation. lambdaRexB inhibits the degradation of the antitoxic labile components Phd and MazE of these systems, which are substrates of ClpP proteases. We present a model for this anti-cell death effect of lambdaRexB through its action on the ClpP proteolytic subunit. We also propose that the lambdarex operon has an additional function to the well known phenomenon of exclusion of other phages; it can prevent the death of lysogenized cells under conditions of nutrient starvation. Thus, the rex operon may be considered as the "survival operon" of phage lambda. PMID- 9860995 TI - The role of mismatch repair in the prevention of base pair mutations in Saccharomyces cerevisiae. AB - In most organisms, the mismatch repair (MMR) system plays an important role in substantially lowering mutation rates and blocking recombination between nonidentical sequences. In Saccharomyces cerevisiae, the products of three genes homologous to Escherichia coli mutS-MSH2, MSH3, and MSH6-function in MMR by recognizing mispaired bases. To determine the effect of MMR on single-base pair mismatches, we have measured reversion rates of specific point mutations in the CYC1 gene in both wild-type and MMR-deficient strains. The reversion rates of all of the point mutations are similar in wild-type cells. However, we find that in the absence of MSH2 or MSH6, but not MSH3, reversion rates of some mutations are increased by up to 60,000-fold, whereas reversion rates of other mutations are essentially unchanged. When cells are grown anaerobically, the reversion rates in MMR-deficient strains are decreased by as much as a factor of 60. We suggest that the high reversion rates observed in these MMR-deficient strains are caused by misincorporations opposite oxidatively damaged bases and that MMR normally prevents these mutations. We further suggest that recognition of mispairs opposite damaged bases may be a more important role for MMR in yeast than correction of errors opposite normal bases. PMID- 9860996 TI - Two functionally dependent acetylcholine subunits are encoded in a single Caenorhabditis elegans operon. AB - The deg-3 gene from the nematode Caenorhabditis elegans encodes an alpha subunit of a nicotinic acetylcholine receptor that was first identified by a dominant allele, u662, which produced neuronal degeneration. Because deg-3 cDNAs contain the SL2 trans-spliced leader, we suggested that deg-3 was transcribed as part of a C. elegans operon. Here we show that des-2, a gene in which mutations suppress deg-3(u662), is the upstream gene in that operon. The des-2 gene also encodes an alpha subunit of a nicotinic acetylcholine receptor. As expected for genes whose mRNAs are formed from a single transcript, both genes have similar expression patterns. This coexpression is functionally important because (i) des-2 is needed for the deg-3(u662) degenerations in vivo; (ii) an acetylcholine-gated channel is formed in Xenopus oocytes when both subunits are expressed but not when either is expressed alone; and (iii) channel activity, albeit apparently altered from that of the wild-type channel, results from the expression of a u662-type mutant subunit but, again, only when the wild-type DES-2 subunit is present. Thus, the operon structure appears to regulate the coordinate expression of two channel subunits. PMID- 9860997 TI - Reduced growth, abnormal kidney structure, and type 2 (AT2) angiotensin receptor mediated blood pressure regulation in mice lacking both AT1A and AT1B receptors for angiotensin II. AB - The classically recognized functions of the renin-angiotensin system are mediated by type 1 (AT1) angiotensin receptors. Whereas man possesses a single AT1 receptor, there are two AT1 receptor isoforms in rodents (AT1A and AT1B) that are products of separate genes (Agtr1a and Agtr1b). We have generated mice lacking AT1B (Agtr1b -/-) and both AT1A and AT1B receptors (Agtr1a -/-Agtr1b -/-). Agtr1b -/- mice are healthy, without an abnormal phenotype. In contrast, Agtr1a -/ Agtr1b -/- mice have diminished growth, vascular thickening within the kidney, and atrophy of the inner renal medulla. This phenotype is virtually identical to that seen in angiotensinogen-deficient (Agt-/-) and angiotensin-converting enzyme deficient (Ace -/-) mice that are unable to synthesize angiotensin II. Agtr1a -/ Agtr1b -/- mice have no systemic pressor response to infusions of angiotensin II, but they respond normally to another vasoconstrictor, epinephrine. Blood pressure is reduced substantially in the Agtr1a -/- Agtr1b -/- mice and following administration of an angiotensin converting enzyme inhibitor, their blood pressure increases paradoxically. We suggest that this is a result of interruption of AT2-receptor signaling. In summary, our studies suggest that both AT1 receptors promote somatic growth and maintenance of normal kidney structure. The absence of either of the AT1 receptor isoforms alone can be compensated in varying degrees by the other isoform. These studies reaffirm and extend the importance of AT1 receptors to mediate physiological functions of the renin angiotensin system. PMID- 9860998 TI - RNA as a target of double-stranded RNA-mediated genetic interference in Caenorhabditis elegans. AB - Introduction of exogenous double-stranded RNA (dsRNA) into Caenorhabditis elegans has been shown to specifically and potently disrupt the activity of genes containing homologous sequences. In this study we present evidence that the primary interference effects of dsRNA are post-transcriptional. First, we examined the primary DNA sequence after dsRNA-mediated interference and found no evidence for alterations. Second, we found that dsRNA-mediated interference with the upstream gene in a polar operon had no effect on the activity of the downstream gene; this finding argues against an effect on initiation or elongation of transcription. Third, we observed by in situ hybridization that dsRNA-mediated interference produced a substantial, although not complete, reduction in accumulation of nascent transcripts in the nucleus, while cytoplasmic accumulation of transcripts was virtually eliminated. These results indicate that the endogenous mRNA is the target for interference and suggest a mechanism that degrades the targeted RNA before translation can occur. This mechanism is not dependent on the SMG system, an mRNA surveillance system in C. elegans responsible for targeting and destroying aberrant messages. We suggest a model of how dsRNA might function in a catalytic mechanism to target homologous mRNAs for degradation. PMID- 9860999 TI - Targeted expression of teashirt induces ectopic eyes in Drosophila. AB - teashirt was initially identified as a gene required for the specification of the trunk segments in Drosophila embryogenesis and encodes a transcription factor with zinc finger motifs. We report here that targeted expression of teashirt in imaginal discs is sufficient to induce ectopic eye formation in non-eye tissues, a phenotype similar to that produced from targeted expression of eyeless, dachshund, and eyes absent. Furthermore, teashirt and eyeless induce the expression of each other, suggesting that teashirt is part of the gene network that functions to specify eye identity. PMID- 9861000 TI - An experimental test for lineage-specific position effects on alcohol dehydrogenase (Adh) genes in Drosophila. AB - Independent transgene insertions differ in expression based on their location in the genome; these position effects are of interest because they reflect the influence of genome organization on gene regulation. Position effects also represent potentially insurmountable obstacles to the rigorous functional comparison of homologous genes from different species because (i) quantitative variation in expression of each gene across genomic positions (generalized position effects, or GPEs) may overwhelm differences between the genes of interest, or (ii) divergent genes may be differentially sensitive to position effects, reflecting unique interactions between each gene and its genomic milieu (lineage-specific position effects, or LSPEs). We have investigated both types of position-effect variation by applying our method of transgene coplacement, which allows comparisons of transgenes in the same position in the genome of Drosophila melanogaster. Here we report an experimental test for LSPE in Drosophila. The alcohol dehydrogenase (Adh) genes of D. melanogaster and Drosophila affinidisjuncta differ in both tissue distribution and amounts of ADH activity. Despite this striking regulatory divergence, we found a very high correlation in overall ADH activity between the genes of the two species when placed in the same genomic position as assayed in otherwise Adh-null adults and larvae. These results argue against the influence of LSPE for these sequences, although the effects of GPE are significant. Our new findings validate the coplacement approach and show that it greatly magnifies the power to detect differences in expression between transgenes. Transgene coplacement thus dramatically extends the range of functional and evolutionary questions that can be addressed by transgenic technology. PMID- 9861001 TI - Efficient translesion replication in the absence of Escherichia coli Umu proteins and 3'-5' exonuclease proofreading function. AB - Translesion replication (TR) past a cyclobutane pyrimidine dimer in Escherichia coli normally requires the UmuD'2C complex, RecA protein, and DNA polymerase III holoenzyme (pol III). However, we find that efficient TR can occur in the absence of the Umu proteins if the 3'-5' exonuclease proofreading activity of the pol III epsilon-subunit also is disabled. TR was measured in isogenic uvrA6 DeltaumuDC strains carrying the dominant negative dnaQ allele, mutD5, or DeltadnaQ spq-2 mutations by transfecting them with single-stranded M13-based vectors containing a specifically located cis-syn T-T dimer. As expected, little TR was observed in the DeltaumuDC dnaQ+ strain. Surprisingly, 26% TR occurred in UV-irradiated DeltaumuDC mutD5 cells, one-half the frequency found in a uvrA6 umuDC+mutD5 strain. lexA3 (Ind-) derivatives of the strains showed that this TR was contingent on two inducible functions, one LexA-dependent, responsible for approximately 70% of the TR, and another LexA-independent, responsible for the remaining approximately 30%. Curiously, the DeltaumuDC DeltadnaQ spq-2 strain exhibited only the LexA-independent level of TR. The cause of this result appears to be the spq-2 allele, a dnaE mutation required for viability in DeltadnaQ strains, since introduction of spq-2 into the DeltaumuDC mutD5 strain also reduces the frequency of TR to the LexA-independent level. The molecular mechanism responsible for the LexA-independent TR is unknown but may be related to the UVM phenomenon [Palejwala, V. A., Wang, G. E., Murphy, H. S. & Humayun, M. Z. (1995) J. Bacteriol. 177, 6041-6048]. LexA-dependent TR does not result from the induction of pol II, since TR in the DeltaumuDC mutD5 strain is unchanged by introduction of a DeltapolB mutation. PMID- 9861002 TI - The JC and BK human polyoma viruses appear to be recent introductions to some South American Indian tribes: there is no serological evidence of cross reactivity with the simian polyoma virus SV40. AB - In an effort to understand the unusual cytogenetic damage earlier encountered in the Yanomama Indians, plasma samples from 425 Amerindians representing 14 tribes have been tested for hemagglutination inhibition antibodies to the human JC polyoma virus and from 369 Amerinds from 13 tribes for hemagglutination inhibition antibodies to the human BK polyoma virus. There is for both viruses highly significant heterogeneity between tribes for the prevalence of serum antibody titers >/=1/40, the pattern of infection suggesting that these two viruses only relatively recently have been introduced into some of these tribes. Some of these samples, from populations with no known exposure to the simian polyoma virus SV40, also were tested for antibodies to this virus by using an immunospot assay. In contrast to the findings of Brown et al. (Brown, P., Tsai, T. & Gajdusek, D. C. (1975) Am. J. Epidemiol. 102, 331-340), none of the samples was found to possess antibodies to SV40. In addition, no significant titers to SV40 were found in a sample of 97 Japanese adults, many of whom had been found to exhibit elevated titers to the JC and BK viruses. This study thus suggests that these human sera contain significant antibody titers to the human polyoma viruses JC and BK but do not appear to contain either cross-reactive antibodies to SV40 or primary antibodies resulting from SV40 infection. PMID- 9861003 TI - A genome-wide search for chromosomal loci linked to mental health wellness in relatives at high risk for bipolar affective disorder among the Old Order Amish. AB - Bipolar affective disorder (BPAD; manic-depressive illness) is characterized by episodes of mania and/or hypomania interspersed with periods of depression. Compelling evidence supports a significant genetic component in the susceptibility to develop BPAD. To date, however, linkage studies have attempted only to identify chromosomal loci that cause or increase the risk of developing BPAD. To determine whether there could be protective alleles that prevent or reduce the risk of developing BPAD, similar to what is observed in other genetic disorders, we used mental health wellness (absence of any psychiatric disorder) as the phenotype in our genome-wide linkage scan of several large multigeneration Old Order Amish pedigrees exhibiting an extremely high incidence of BPAD. We have found strong evidence for a locus on chromosome 4p at D4S2949 (maximum GENEHUNTER PLUS nonparametric linkage score = 4.05, P = 5. 22 x 10(-4); SIBPAL Pempirical value <3 x 10(-5)) and suggestive evidence for a locus on chromosome 4q at D4S397 (maximum GENEHUNTER-PLUS nonparametric linkage score = 3.29, P = 2.57 x 10(-3); SIBPAL Pempirical value <1 x 10(-3)) that are linked to mental health wellness. These findings are consistent with the hypothesis that certain alleles could prevent or modify the clinical manifestations of BPAD and perhaps other related affective disorders. PMID- 9861004 TI - A potential role for the nuclear factor of activated T cells family of transcriptional regulatory proteins in adipogenesis. AB - NFAT (nuclear factor of activated T cells) is a family of transcription factors implicated in the control of cytokine and early immune response gene expression. Recent studies have pointed to a role for NFAT proteins in gene regulation outside of the immune system. Herein we demonstrate that NFAT proteins are present in 3T3-L1 adipocytes and, upon fat cell differentiation, bind to and transactivate the promoter of the adipocyte-specific gene aP2. Further, fat cell differentiation is inhibited by cyclosporin A, a drug shown to prevent NFAT nuclear localization and hence function. Thus, these data suggest a role for NFAT transcription factors in the regulation of the aP2 gene and in the process of adipocyte differentiation. PMID- 9861005 TI - Anti-beta2 glycoprotein I (beta2GPI) autoantibodies recognize an epitope on the first domain of beta2GPI. AB - Anticardiolipin (aCL) autoantibodies are associated with thrombosis, recurrent fetal loss, and thrombocytopenia. Only aCL found in autoimmune disease require the participation of the phospholipid binding plasma protein beta2 glycoprotein I (beta2GPI) for antibody binding and now are called anti-beta2GPI. The antigenic specificity of aCL affinity purified from 11 patients with high titers was evaluated in an effort to better understand the pathophysiology associated with aCL. Seven different recombinant domain-deleted mutants of human beta2GPI, and full length human beta2GPI (wild-type), were used in competition assays to inhibit the autoantibodies from binding to immobilized wild-type beta2GPI. Only those domain-deleted mutants that contained domain 1 inhibited the binding to immobilized wild-type beta2GPI from all of the patients. The domain-deleted mutants that contained domain 1 inhibited all aCL in a similar but not identical pattern, suggesting that these aCL recognize a similar, but distinguishable, epitope(s) present on domain 1. PMID- 9861006 TI - Altered thymic positive selection and intracellular signals in Cbl-deficient mice. AB - Cbl is the product of the protooncogene c-cbl and is involved in T cell antigen receptor (TCR)-mediated signaling. To understand the role of Cbl for immune system development and function, we generated a Cbl-deficient mouse strain. In Cbl-deficient mice, positive selection of the thymocytes expressing major histocompatibility complex class II-restricted transgenic TCR was significantly enhanced. Two factors may have contributed to the altered thymic selection. First, Cbl deficiency markedly up-regulated the activity of ZAP-70 and mitogen activated protein kinases. The mitogen-activated protein kinase pathway was shown previously to be involved in thymic positive selection. Second, Cbl-deficient thymocytes expressed CD3 and CD4 molecules at higher levels, which consequently may increase the avidity of TCR/major histocompatibility complex/coreceptor interaction. Thus, Cbl plays a novel role in modulating TCR-mediated multiple signaling pathways and fine-tunes the signaling threshold for thymic selection. PMID- 9861008 TI - Analysis of variable (diversity) joining recombination in DNAdependent protein kinase (DNA-PK)-deficient mice reveals DNA-PK-independent pathways for both signal and coding joint formation. AB - Previous studies have suggested that ionizing radiation causes irreparable DNA double-strand breaks in mice and cell lines harboring mutations in any of the three subunits of DNA-dependent protein kinase (DNA-PK) (the catalytic subunit, DNA-PKcs, or one of the DNA-binding subunits, Ku70 or Ku86). In actuality, these mutants vary in their ability to resolve double-strand breaks generated during variable (diversity) joining [V(D)J] recombination. Mutant cell lines and mice with targeted deletions in Ku70 or Ku86 are severely compromised in their ability to form coding and signal joints, the products of V(D)J recombination. It is noteworthy, however, that severe combined immunodeficient (SCID) mice, which bear a nonnull mutation in DNA-PKcs, are substantially less impaired in forming signal joints than coding joints. The current view holds that the defective protein encoded by the murine SCID allele retains enough residual function to support signal joint formation. An alternative hypothesis proposes that DNA-PKcs and Ku perform different roles in V(D)J recombination, with DNA-PKcs required only for coding joint formation. To resolve this issue, we examined V(D)J recombination in DNA-PKcs-deficient (SLIP) mice. We found that the effects of this mutation on coding and signal joint formation are identical to the effects of the SCID mutation. Signal joints are formed at levels 10-fold lower than in wild type, and one-half of these joints are aberrant. These data are incompatible with the notion that signal joint formation in SCID mice results from residual DNA-PKcs function, and suggest a third possibility: that DNA-PKcs normally plays an important but nonessential role in signal joint formation. PMID- 9861007 TI - CpG DNA can induce strong Th1 humoral and cell-mediated immune responses against hepatitis B surface antigen in young mice. AB - Successful neonatal immunization of humans has proven difficult. We have evaluated CpG-containing oligonucleotides as an adjuvant for immunization of young mice (1-14 days old) against hepatitis B virus surface antigen. The protein alum-CpG formulation, like the DNA vaccine, produced seroconversion of the majority of mice immunized at 3 or 7 days of age, compared with 0-10% with the protein-alum or protein-CpG formulations. All animals, from neonates to adults, immunized with the protein-alum vaccine exhibited strong T helper (Th)2-like responses [predominantly IgG1, weak or absent cytotoxic T lymphocytes (CTL)]. Th2 type responses also were induced in young mice with protein-CpG (in 1-, 3-, and 7 day-old mice) and protein-alum-CpG (in 1- and 3-day-old mice) but immunization carried out at older ages gave mixed Th1/Th2 (Th0) responses. DNA vaccines gave Th0-like responses when administered at 1 and 7 days of age and Th1-like (predominantly IgG2a and CTL) responses with 14-day-old or adult mice. Surprisingly, the protein-alum-CpG formulation was better than the DNA vaccine for percentage of seroconversion, speed of appearance, and peak titer of the antibody response, as well as prevalence and strength of CTL. These findings may have important implications for immunization of human infants. PMID- 9861009 TI - Virus-specific CD8(+) T cell numbers are maintained during gamma-herpesvirus reactivation in CD4-deficient mice. AB - The murine gamma-herpesvirus 68 replicates in epithelial sites after intranasal challenge, then persists in various cell types, including B lymphocytes. Mice that lack CD4(+) T cells (I-Ab-/-) control the acute infection, but suffer an ultimately lethal recrudescence of lytic viral replication in the respiratory tract. The consequences of CD4(+) T cell deficiency for the generation and maintenance of murine gamma-herpesvirus 68-specific CD8(+) set now have been analyzed by direct staining with viral peptides bound to major histocompatibility complex class I tetramers and by a spectrum of functional assays. Both acutely and during viral reactivation, the CD8(+) T cell responses in the I-Ab-/- group were no less substantial than in the I-Ab+/+ controls. Indeed, virus-specific CD8(+) T cell numbers were increased in the lymphoid tissue of clinically compromised I-Ab-/- mice, although relatively few of the potential cytotoxic T lymphocyte effectors were recruited back to the site of pathology in the lung. Thus the viral reactivation that occurs in the absence of CD4(+) T cells was not associated with any exhaustion of the virus-specific cytotoxic T lymphocyte response. It seems that CD8(+) T cells alone are insufficient to maintain long term control of this persistent gamma-herpesvirus. PMID- 9861010 TI - Variant antigenic peptide promotes cytotoxic T lymphocyte adhesion to target cells without cytotoxicity. AB - Timelapse video microscopy has been used to record the motility and dynamic interactions between an H-2Db-restricted murine cytotoxic T lymphocyte clone (F5) and Db-transfected L929 mouse fibroblasts (LDb) presenting normal or variant antigenic peptides from human influenza nucleoprotein. F5 cells will kill LDb target cells presenting specific antigen (peptide NP68: ASNENMDAM) after "browsing" their surfaces for between 8 min and many hours. Cell death is characterized by abrupt cellular rounding followed by zeiosis (vigorous "boiling" of the cytoplasm and blebbing of the plasma membrane) for 10-20 min, with subsequent cessation of all activity. Departure of cytotoxic T lymphocytes from unkilled target cells is rare, whereas serial killing is sometimes observed. In the absence of antigenic peptide, cytotoxic T lymphocytes browse target cells for much shorter periods, and readily leave to encounter other targets, while never causing target cell death. Two variant antigenic peptides, differing in nonamer position 7 or 8, also act as antigens, albeit with lower efficiency. A third variant peptide NP34 (ASNENMETM), which differs from NP68 in both positions and yet still binds Db, does not stimulate F5 cytotoxicity. Nevertheless, timelapse video analysis shows that NP34 leads to a significant modification of cell behavior, by up-regulating F5-LDb adhesive interactions. These data extend recent studies showing that partial agonists may elicit a subset of the T cell responses associated with full antigen stimulation, by demonstrating that TCR interaction with variant peptide antigens can trigger target cell adhesion and surface exploration without activating the signaling pathway that results in cytotoxicity. PMID- 9861011 TI - Accelerated apoptosis of lymphocytes by augmented induction of Bax in SSI-1 (STAT induced STAT inhibitor-1) deficient mice. AB - Growth, differentiation, and programmed cell death (apoptosis) are mainly controlled by cytokines. The Janus kinase-signal transducers and activators of transcription (JAK-STAT) signal pathway is an important component of cytokine signaling. We have previously shown that STAT3 induces a molecule designated as SSI-1, which inhibits STAT3 functions. To clarify the physiological roles of SSI 1 in vivo, we generated, here, mice lacking SSI-1. These SSI-1-/- mice displayed growth retardation and died within 3 weeks after birth. Lymphocytes in the thymus and spleen of the SSI-1-/- mice exhibited accelerated apoptosis with aging, and their number was 20-25% of that in SSI-1+/+ mice at 10 days of age. However, the differentiation of lymphocytes lacking SSI-1 appeared to be normal. Among various pro- and anti-apoptotic molecules examined, an up-regulation of Bax was found in lymphocytes of the spleen and thymus of SSI-1-/- mice. These findings suggest that SSI-1 prevents apoptosis by inhibiting the expression of Bax. PMID- 9861012 TI - Endogenous E2F-1 promotes timely G0 exit of resting mouse embryo fibroblasts. AB - Much evidence strongly suggests a positive role for one or more E2F species in the control of exit from G0/G1. Results described here provide direct evidence that endogenous E2F-1, as predicted, contributes to progression from G0 to S. By contrast, cycling cells lacking an intact E2F-1 gene demonstrated normal cell cycle distribution. Therefore, E2F-1 exerts a unique function leading to timely G0 exit of resting cultured primary cells, while at the same time being unnecessary for normal G1 to S phase progression of cycling cells. PMID- 9861013 TI - The PTEN/MMAC1 tumor suppressor phosphatase functions as a negative regulator of the phosphoinositide 3-kinase/Akt pathway. AB - The PTEN/MMAC1 phosphatase is a tumor suppressor gene implicated in a wide range of human cancers. Here we provide biochemical and functional evidence that PTEN/MMAC1 acts a negative regulator of the phosphoinositide 3-kinase (PI3 kinase)/Akt pathway. PTEN/MMAC1 impairs activation of endogenous Akt in cells and inhibits phosphorylation of 4E-BP1, a downstream target of the PI3-kinase/Akt pathway involved in protein translation, whereas a catalytically inactive, dominant negative PTEN/MMAC1 mutant enhances 4E-BP1 phosphorylation. In addition, PTEN/MMAC1 represses gene expression in a manner that is rescued by Akt but not PI3-kinase. Finally, higher levels of Akt activation are observed in human prostate cancer cell lines and xenografts lacking PTEN/MMAC1 expression when compared with PTEN/MMAC1-positive prostate tumors or normal prostate tissue. Because constitutive activation of either PI3-kinase or Akt is known to induce cellular transformation, an increase in the activation of this pathway caused by mutations in PTEN/MMAC1 provides a potential mechanism for its tumor suppressor function. PMID- 9861014 TI - Targeted disruption of mouse long-chain acyl-CoA dehydrogenase gene reveals crucial roles for fatty acid oxidation. AB - Abnormalities of fatty acid metabolism are recognized to play a significant role in human disease, but the mechanisms remain poorly understood. Long-chain acyl CoA dehydrogenase (LCAD) catalyzes the initial step in mitochondrial fatty acid oxidation (FAO). We produced a mouse model of LCAD deficiency with severely impaired FAO. Matings between LCAD +/- mice yielded an abnormally low number of LCAD +/- and -/- offspring, indicating frequent gestational loss. LCAD -/- mice that reached birth appeared normal, but had severely reduced fasting tolerance with hepatic and cardiac lipidosis, hypoglycemia, elevated serum free fatty acids, and nonketotic dicarboxylic aciduria. Approximately 10% of adult LCAD -/- males developed cardiomyopathy, and sudden death was observed in 4 of 75 LCAD -/- mice. These results demonstrate the crucial roles of mitochondrial FAO and LCAD in vivo. PMID- 9861015 TI - Opposite roles of apolipoprotein E in normal brains and in Alzheimer's disease. AB - We have characterized the interaction between apolipoprotein E (apoE) and amyloid beta peptide (Abeta) in the soluble fraction of the cerebral cortex of Alzheimer's disease (AD) and control subjects. Western blot analysis with specific antibodies identified in both groups a complex composed of the full length apoE and Abeta peptides ending at residues 40 and 42. The apoE-Abeta soluble aggregate is less stable in AD brains than in controls, when treated with the anionic detergent SDS. The complex is present in significantly higher quantity in control than in AD brains, whereas in the insoluble fraction an inverse correlation has previously been reported. Moreover, in the AD subjects the Abeta bound to apoE is more sensitive to protease digestion than is the unbound Abeta. Taken together, our results indicate that in normal brains apoE efficiently binds and sequesters Abeta, preventing its aggregation. In AD, the impaired apoE-Abeta binding leads to the critical accumulation of Abeta, facilitating plaque formation. PMID- 9861016 TI - Viral mediated expression of insulin-like growth factor I blocks the aging related loss of skeletal muscle function. AB - During the aging process, mammals lose up to a third of their skeletal muscle mass and strength. Although the mechanisms underlying this loss are not entirely understood, we attempted to moderate the loss by increasing the regenerative capacity of muscle. This involved the injection of a recombinant adeno-associated virus directing overexpression of insulin-like growth factor I (IGF-I) in differentiated muscle fibers. We demonstrate that the IGF-I expression promotes an average increase of 15% in muscle mass and a 14% increase in strength in young adult mice, and remarkably, prevents aging-related muscle changes in old adult mice, resulting in a 27% increase in strength as compared with uninjected old muscles. Muscle mass and fiber type distributions were maintained at levels similar to those in young adults. We propose that these effects are primarily due to stimulation of muscle regeneration via the activation of satellite cells by IGF-I. This supports the hypothesis that the primary cause of aging-related impairment of muscle function is a cumulative failure to repair damage sustained during muscle utilization. Our results suggest that gene transfer of IGF-I into muscle could form the basis of a human gene therapy for preventing the loss of muscle function associated with aging and may be of benefit in diseases where the rate of damage to skeletal muscle is accelerated. PMID- 9861017 TI - Overexpression of Mdm2 in mice reveals a p53-independent role for Mdm2 in tumorigenesis. AB - The Mdm2 proto-oncogene is amplified to high copy numbers in human sarcomas and is overexpressed in a wide variety of other human cancers. Because Mdm2 protein forms a complex with the p53 tumor suppressor protein and down-regulates p53 function, the oncogenic potential of Mdm2 is presumed to be p53-dependent. To model these conditions in mice, we have used the entire Mdm2 gene, under transcriptional control of its native promoter region, as a transgene to create mice that overexpress Mdm2. The transgenic mice are predisposed to spontaneous tumor formation, and the incidence of sarcomas observed in the Mdm2-transgenic mice in the presence or absence of functional p53 demonstrates that, in addition to Mdm2-mediated inactivation of p53, there exists a p53-independent role for Mdm2 in tumorigenesis. PMID- 9861018 TI - Evidence that a prominent cavity in the coiled coil of HIV type 1 gp41 is an attractive drug target. AB - Synthetic C peptides, corresponding to the C helix of the HIV type 1 (HIV-1) gp41 envelope protein, are potent inhibitors of HIV-1 membrane fusion. One such peptide is in clinical trials. The crystal structure of the gp41 core, in its proposed fusion-active conformation, is a trimer of helical hairpins in which three C helices pack against a central coiled coil. Each C helix shows especially prominent contacts with one of three symmetry-related, hydrophobic cavities on the surface of the coiled coil. We show that the inhibitory activity of the C peptide C34 depends on its ability to bind to this coiled-coil cavity. Moreover, examining a series of C34 peptide variants with modified cavity-binding residues, we find a linear relationship between the logarithm of the inhibitory potency and the stability of the corresponding helical-hairpin complexes. Our results provide strong evidence that this coiled-coil cavity is a good drug target and clarify the mechanism of C peptide inhibition. They also suggest simple, quantitative assays for the identification and evaluation of analogous inhibitors of HIV-1 entry. PMID- 9861019 TI - The effects of health changes on projections of health service needs for the elderly population of the United States. AB - The 1982-1994 National Long-Term Care Surveys indicate an accelerating decline in disability among the U.S. elderly population, suggesting that a 1.5% annual decline in chronic disability for elderly persons is achievable. Furthermore, many risk factors for chronic diseases show improvements, many linked to education, from 1910 to the present. Projections indicate the proportion of persons aged 85-89 with less than 8 years of education will decline from 65% in 1980 to 15% in 2015. Health and socioeconomic status trends are not directly represented in Medicare Trust Fund and Social Security Administration beneficiary projections. Thus, they may have different economic implications from projections directly accounting for health trends. A 1.5% annual disability decline keeps the support ratio (ratio of economically active persons aged 20-64 to the number of chronically disabled persons aged 65+) above its 1994 value, 22:1, when the Hospital Insurance Trust Fund was in fiscal balance, to 2070. With no changes in disability, projections indicate a support ratio in 2070 of 8:1-63% below a cash flow balance. PMID- 9861020 TI - Identification of genes differentially regulated by interferon alpha, beta, or gamma using oligonucleotide arrays. AB - The pleiotropic activities of interferons (IFNs) are mediated primarily through the transcriptional regulation of many downstream effector genes. The mRNA profiles from IFN-alpha, -beta, or -gamma treatments of the human fibrosarcoma cell line, HT1080, were determined by using oligonucleotide arrays with probe sets corresponding to more than 6,800 human genes. Among these were transcripts for known IFN-stimulated genes (ISGs), the expression of which were consistent with previous studies in which the particular ISG was characterized as responsive to either Type I (alpha, beta) or Type II (gamma) IFNs, or both. Importantly, many novel IFN-stimulated genes were identified that were diverse in their known biological functions. For instance, several novel ISGs were identified that are implicated in apoptosis (including RAP46/Bag-1, phospholipid scramblase, and hypoxia inducible factor-1alpha). Furthermore, several IFN-repressed genes also were identified. These results demonstrate the usefulness of oligonucleotide arrays in monitoring mammalian gene expression on a broad and unprecedented scale. In particular, these findings provide insights into the basic mechanisms of IFN actions and ultimately may contribute to better therapeutic uses for IFNs. PMID- 9861021 TI - Loss of function of the tuberous sclerosis 2 tumor suppressor gene results in embryonic lethality characterized by disrupted neuroepithelial growth and development. AB - Germline defects in the tuberous sclerosis 2 (TSC2) tumor suppressor gene predispose humans and rats to benign and malignant lesions in a variety of tissues. The brain is among the most profoundly affected organs in tuberous sclerosis (TSC) patients and is the site of development of the cortical tubers for which the hereditary syndrome is named. A spontaneous germline inactivation of the Tsc2 locus has been described in an animal model, the Eker rat. We report that the homozygous state of this mutation (Tsc2(Ek/Ek)) was lethal in mid gestation (the equivalent of mouse E9.5-E13.5), when Tsc2 mRNA was highly expressed in embryonic neuroepithelium. During this period homozygous mutant Eker embryos lacking functional Tsc2 gene product, tuberin, displayed dysraphia and papillary overgrowth of the neuroepithelium, indicating that loss of tuberin disrupted the normal development of this tissue. Interestingly, there was significant intraspecies variability in the penetrance of cranial abnormalities in mutant embryos: the Long-Evans strain Tsc2(Ek/Ek) embryos displayed these defects whereas the Fisher 344 homozygous mutant embryos had normal-appearing neuroepithelium. Taken together, our data indicate that the Tsc2 gene participates in normal brain development and suggest the inactivation of this gene may have similar functional consequences in both mature and embryonic brain. PMID- 9861022 TI - Repetitive transcranial magnetic stimulation activates specific regions in rat brain. AB - Repetitive transcranial magnetic stimulation (rTMS) is a noninvasive technique to induce electric currents in the brain. Although rTMS is being evaluated as a possible alternative to electroconvulsive therapy for the treatment of refractory depression, little is known about the pattern of activation induced in the brain by rTMS. We have compared immediate early gene expression in rat brain after rTMS and electroconvulsive stimulation, a well-established animal model for electroconvulsive therapy. Our result shows that rTMS applied in conditions effective in animal models of depression induces different patterns of immediate early gene expression than does electroconvulsive stimulation. In particular, rTMS evokes strong neural responses in the paraventricular nucleus of the thalamus (PVT) and in other regions involved in the regulation of circadian rhythms. The response in PVT is independent of the orientation of the stimulation probe relative to the head. Part of this response is likely because of direct activation, as repetitive magnetic stimulation also activates PVT neurons in brain slices. PMID- 9861023 TI - Modeling stochastic gene expression: implications for haploinsufficiency. AB - There is increasing recognition that stochastic processes regulate highly predictable patterns of gene expression in developing organisms, but the implications of stochastic gene expression for understanding haploinsufficiency remain largely unexplored. We have used simulations of stochastic gene expression to illustrate that gene copy number and expression deactivation rates are important variables in achieving predictable outcomes. In gene expression systems with non-zero expression deactivation rates, diploid systems had a higher probability of uninterrupted gene expression than haploid systems and were more successful at maintaining gene product above a very low threshold. Systems with relatively rapid expression deactivation rates (unstable gene expression) had more predictable responses to a gradient of inducer than systems with slow or zero expression deactivation rates (stable gene expression), and diploid systems were more predictable than haploid, with or without dosage compensation. We suggest that null mutations of a single allele in a diploid organism could decrease the probability of gene expression and present the hypothesis that some haploinsufficiency syndromes might result from an increased susceptibility to stochastic delays of gene initiation or interruptions of gene expression. PMID- 9861024 TI - Lamellar lipoproteins uniquely contribute to hyperlipidemia in mice doubly deficient in apolipoprotein E and hepatic lipase. AB - Remnants of triglyceride-rich lipoproteins containing apolipoprotein (apo) B-48 accumulate in apo E-deficient mice, causing pronounced hypercholesterolemia. Mice doubly deficient in apo E and hepatic lipase have more pronounced hypercholesterolemia, even though remnants do not accumulate appreciably in mice deficient in hepatic lipase alone. Here we show that the doubly deficient mice manifest a unique lamellar hyperlipoproteinemia, characterized by vesicular particles 600 A-1,300 A in diameter. As seen by negative-staining electron microscopy, these lipoproteins also contain an electron-lucent region adjacent to the vesicle wall, similar to the core of typical lipoproteins. Correlative chemical analysis indicates that the vesicle wall is composed of a 1:1 molar mixture of cholesterol and phospholipids, whereas the electron-lucent region appears to be composed of cholesteryl esters (about 12% of the particle mass). Like the spherical lipoproteins of doubly deficient mice, the vesicular particles contain apo B-48, but they are particularly rich in apo A-IV. We propose that cholesteryl esters are removed from spherical lipoproteins of these mice by scavenger receptor B1, leaving behind polar lipid-rich particles that fuse to form vesicular lipoproteins. Hepatic lipase may prevent such vesicular lipoproteins from accumulating in apo E-deficient mice by hydrolyzing phosphatidyl choline as scavenger receptor B1 removes the cholesteryl esters and by gradual endocytosis of lipoproteins bound to hepatic lipase on the surface of hepatocytes. PMID- 9861025 TI - Inactivation of the cyclin-dependent kinase inhibitor p27 upon loss of the tuberous sclerosis complex gene-2. AB - Tuberous sclerosis is an autosomal dominant disorder characterized by the development of aberrant growths in many tissues and organs. Linkage analysis revealed two disease-determining genes on chromosome 9 and chromosome 16. The tuberous sclerosis complex gene-2 (TSC2) on chromosome 16 encodes the tumor suppressor protein tuberin. We have shown earlier that loss of TSC2 is sufficient to induce quiescent cells to enter the cell cycle. Here we show that TSC2 negative fibroblasts exhibit a shortened G1 phase. Although the expression of cyclin E, cyclin A, p21, or Cdc25A is unaffected, TSC2-negative cells express much lower amounts of the cyclin-dependent kinase (CDK) inhibitor p27 because of decreased protein stability. In TSC2 mutant cells the amount of p27 bound to CDK2 is diminished, accompanied with elevated kinase activity. Ectopic expression studies revealed that the aforementioned effects can be reverted by transfecting TSC2 in TSC2-negative cells. High ectopic levels of p27 have cell cycle inhibitory effects in TSC2-positive cells but not in TSC2-negative counterparts, although the latter still depend on CDK2 activity. Loss of TSC2 induces soft agar growth of fibroblasts, a process that cannot be inhibited by high levels of p27. Both phenotypes of TSC2-negative cells, their resistance to the activity of ectopic p27, and the instability of endogenous p27, could be explained by our observation that the nucleoprotein p27 is mislocated into the cytoplasm upon loss of TSC2. These findings provide insights into the molecular mechanism of how loss of TSC2 induces cell cycle entry and allow a better understanding of its tumor suppressor function. PMID- 9861026 TI - Obesity and mild hyperinsulinemia found in neuropeptide Y-Y1 receptor-deficient mice. AB - To elucidate the role of neuropeptide Y (NPY)-Y1 receptor (Y1-R) in food intake, energy expenditure, and other possible functions, we have generated Y1-R deficient mice (Y1-R-/-) by gene targeting. Contrary to our hypothesis that the lack of NPY signaling via Y1-R would result in impaired feeding and weight loss, Y1-R-/- mice showed a moderate obesity and mild hyperinsulinemia without hyperphagia. Although there was some variation between males and females, typical characteristics of Y1-R-/- mice include: greater body weight (females more than males), an increase in the weight of white adipose tissue (WAT) (approximately 4 fold in females), an elevated basal level of plasma insulin (approximately 2 fold), impaired insulin secretion in response to glucose administration, and a significant changes in mitochondrial uncoupling protein (UCP) gene expression (up regulation of UCP1 in brown adipose tissue and down-regulation of UCP2 in WAT). These results suggest either that the Y1-R in the hypothalamus is not a key molecule in the leptin/NPY pathway, which controls feeding behavior, or that its deficiency is compensated by other receptors, such as NPY-Y5 receptor. We believe that the mild obesity found in Y1-R-/- mice (especially females) was caused by the impaired control of insulin secretion and/or low energy expenditure, including the lowered expression of UCP2 in WAT. This model will be useful for studying the mechanism of mild obesity and abnormal insulin metabolism in noninsulin-dependent diabetes mellitus. PMID- 9861027 TI - A multidrug resistance transporter from human MCF-7 breast cancer cells. AB - MCF-7/AdrVp is a multidrug-resistant human breast cancer subline that displays an ATP-dependent reduction in the intracellular accumulation of anthracycline anticancer drugs in the absence of overexpression of known multidrug resistance transporters such as P glycoprotein or the multidrug resistance protein. RNA fingerprinting led to the identification of a 2.4-kb mRNA that is overexpressed in MCF-7/AdrVp cells relative to parental MCF-7 cells. The mRNA encodes a 655-aa [corrected] member of the ATP-binding cassette superfamily of transporters that we term breast cancer resistance protein (BCRP). Enforced expression of the full length BCRP cDNA in MCF-7 breast cancer cells confers resistance to mitoxantrone, doxorubicin, and daunorubicin, reduces daunorubicin accumulation and retention, and causes an ATP-dependent enhancement of the efflux of rhodamine 123 in the cloned transfected cells. BCRP is a xenobiotic transporter that appears to play a major role in the multidrug resistance phenotype of MCF-7/AdrVp human breast cancer cells. PMID- 9861029 TI - Generation and reproductive phenotypes of mice lacking estrogen receptor beta. AB - Estrogens influence the differentiation and maintenance of reproductive tissues and affect lipid metabolism and bone remodeling. Two estrogen receptors (ERs) have been identified to date, ERalpha and ERbeta. We previously generated and studied knockout mice lacking estrogen receptor alpha and reported severe reproductive and behavioral phenotypes including complete infertility of both male and female mice and absence of breast tissue development. Here we describe the generation of mice lacking estrogen receptor beta (ERbeta -/-) by insertion of a neomycin resistance gene into exon 3 of the coding gene by using homologous recombination in embryonic stem cells. Mice lacking this receptor develop normally and are indistinguishable grossly and histologically as young adults from their littermates. RNA analysis and immunocytochemistry show that tissues from ERbeta -/- mice lack normal ERbeta RNA and protein. Breeding experiments with young, sexually mature females show that they are fertile and exhibit normal sexual behavior, but have fewer and smaller litters than wild-type mice. Superovulation experiments indicate that this reduction in fertility is the result of reduced ovarian efficiency. The mutant females have normal breast development and lactate normally. Young, sexually mature male mice show no overt abnormalities and reproduce normally. Older mutant males display signs of prostate and bladder hyperplasia. Our results indicate that ERbeta is essential for normal ovulation efficiency but is not essential for female or male sexual differentiation, fertility, or lactation. Future experiments are required to determine the role of ERbeta in bone and cardiovascular homeostasis. PMID- 9861028 TI - Role of the proteasome and NF-kappaB in streptococcal cell wall-induced polyarthritis. AB - The transcription factor NF-kappaB activates a number of genes whose protein products are proinflammatory. In quiescent cells, NF-kappaB exists in a latent form and is activated via a signal-dependent proteolytic mechanism in which the inhibitory protein IkappaB is degraded by the ubiquitin-proteasome pathway. Consequently, inhibition of the proteasome suppresses activation of NF-kappaB. This suppression should therefore decrease transcription of many genes encoding proinflammatory proteins and should ultimately have an anti-inflammatory effect. To this end, a series of peptide boronic acid inhibitors of the proteasome, exemplified herein by PS-341, were developed. The proteasome is the large multimeric protease that catalyzes the final proteolytic step of the ubiquitin proteasome pathway. PS-341, a potent, competitive inhibitor of the proteasome, readily entered cells and inhibited the activation of NF-kappaB and the subsequent transcription of genes that are regulated by NF-kappaB. Significantly, PS-341 displayed similar effects in vivo. Oral administration of PS-341 had anti inflammatory effects in a model of Streptococcal cell wall-induced polyarthritis and liver inflammation in rats. The attenuation of inflammation in this model was associated with an inhibition of IkappaBalpha degradation and NF-kappaB-dependent gene expression. These experiments clearly demonstrate that the ubiquitin proteasome pathway and NF-kappaB play important roles in regulating chronic inflammation and that, as predicted, proteasome inhibition has an anti inflammatory effect. PMID- 9861030 TI - Adenovirus E4orf6 oncoprotein modulates the function of the p53-related protein, p73. AB - Recently, several proteins have been identified that are related in their sequence to the p53 tumor-suppressor protein. One of these proteins, which is termed p73, exhibits sequence homology to the p53 transcriptional activation, DNA binding, and oligomerization domains. The adenovirus E1B 55-kDa protein, the adenovirus E4orf6 protein, and SV40 T antigen each can bind to p53 and inhibit p53 function. Here we demonstrate that the adenovirus E4orf6 protein, but not the E1B 55-kDa protein or T antigen, interacts with p73. The E4orf6 protein inhibits p73-mediated transcriptional activation and cell killing in a manner similar to its effect on p53. Thus, only a subset of viral oncoproteins that antagonize p53 function also interacts with the related p73 protein. PMID- 9861031 TI - A mAb recognizing a surface antigen of Mycobacterium tuberculosis enhances host survival. AB - Murine mAbs reactive with the surface of Mycobacterium tuberculosis were assayed for their ability to affect the course of infection in mice challenged with virulent organisms. An IgG3 mAb (9d8) specific for arabinomannan and reactive with purified antigen from a clinical isolate of M. tuberculosis conferred partial protection on mice after respiratory challenge (30-60% survival >75 days; P 35,000-fold more potent than paclitaxel in inhibiting cell growth in the MDR DC-3F/ADX cell line. Various formulations, routes, and schedules of i.v. administration of dEpoB have been tested in nude mice. Slow infusion with a Cremophor-ethanol vehicle proved to be the most beneficial in increasing efficacy and decreasing toxicity. Although dEpoB performed similarly to paclitaxel in sensitive tumors xenografts (MX-1 human mammary and HT-29 colon tumor), its effects were clearly superior against MDR tumors. When dEpoB was administered to nude mice bearing our MDR human lymphoblastic T cell leukemia (CCRF CEM/paclitaxel), dEpoB demonstrated a full curative effect. For human mammary adenocarcinoma MCF-7/Adr cells refractory to paclitaxel, dEpoB reduced the established tumors, markedly suppressed tumor growth, and surpassed other commonly used chemotherapy drugs such as adriamycin, vinblastine, and etoposide in beneficial effects. PMID- 9861051 TI - Cloning of the cDNA encoding the urotensin II precursor in frog and human reveals intense expression of the urotensin II gene in motoneurons of the spinal cord. AB - Urotensin II (UII) is a cyclic peptide initially isolated from the caudal neurosecretory system of teleost fish. Subsequently, UII has been characterized from a frog brain extract, indicating that a gene encoding a UII precursor is also present in the genome of a tetrapod. Here, we report the characterization of the cDNAs encoding frog and human UII precursors and the localization of the corresponding mRNAs. In both frog and human, the UII sequence is located at the C terminal position of the precursor. Human UII is composed of only 11 amino acid residues, while fish and frog UII possess 12 and 13 amino acid residues, respectively. The cyclic region of UII, which is responsible for the biological activity of the peptide, has been fully conserved from fish to human. Northern blot and dot blot analysis revealed that UII precursor mRNAs are found predominantly in the frog and human spinal cord. In situ hybridization studies showed that the UII precursor gene is actively expressed in motoneurons. The present study demonstrates that UII, which has long been regarded as a peptide exclusively produced by the urophysis of teleost fish, is actually present in the brain of amphibians and mammals. The fact that evolutionary pressure has acted to conserve fully the biologically active sequence of UII suggests that the peptide may exert important physiological functions in humans. PMID- 9861052 TI - Systemic hypoxia changes the organ-specific distribution of vascular endothelial growth factor and its receptors. AB - Vascular endothelial growth factor (VEGF) plays a key role in physiological blood vessel formation and pathological angiogenesis such as tumor growth and ischemic diseases. Hypoxia is a potent inducer of VEGF in vitro. Here we demonstrate that VEGF is induced in vivo by exposing mice to systemic hypoxia. VEGF induction was highest in brain, but also occurred in kidney, testis, lung, heart, and liver. In situ hybridization analysis revealed that a distinct subset of cells within a given organ, such as glial cells and neurons in brain, tubular cells in kidney, and Sertoli cells in testis, responded to the hypoxic stimulus with an increase in VEGF expression. Surprisingly, however, other cells at sites of constitutive VEGF expression in normal adult tissues, such as epithelial cells in the choroid plexus and kidney glomeruli, decreased VEGF expression in response to the hypoxic stimulus. Furthermore, in addition to VEGF itself, expression of VEGF receptor-1 (VEGFR-1), but not VEGFR-2, was induced by hypoxia in endothelial cells of lung, heart, brain, kidney, and liver. VEGF itself was never found to be up-regulated in endothelial cells under hypoxic conditions, consistent with its paracrine action during normoxia. Our results show that the response to hypoxia in vivo is differentially regulated at the level of specific cell types or layers in certain organs. In these tissues, up- or down-regulation of VEGF and VEGFR-1 during hypoxia may influence their oxygenation after angiogenesis or modulate vascular permeability. PMID- 9861053 TI - The effect of 4,4'-diisothiocyanato-stilbene-2,2'-disulfonate on CO2 permeability of the red blood cell membrane. AB - It has long been assumed that the red cell membrane is highly permeable to gases because the molecules of gases are small, uncharged, and soluble in lipids, such as those of a bilayer. The disappearance of 12C18O16O from a red cell suspension as the 18O exchanges between labeled CO2 + HCO3- and unlabeled HOH provides a measure of the carbonic anhydrase (CA) activity (acceleration, or A) inside the cell and of the membrane self-exchange permeability to HCO3- (Pm,HCO-3). To test this technique, we added sufficient 4, 4'-diisothiocyanato-stilbene-2,2' disulfonate (DIDS) to inhibit all the HCO3-/Cl- transport protein (Band III or capnophorin) in a red cell suspension. We found that DIDS reduced Pm,HCO-3 as expected, but also appeared to reduce intracellular A, although separate experiments showed it has no effect on CA activity in homogenous solution. A decrease in Pm,CO2 would explain this finding. With a more advanced computational model, which solves for CA activity and membrane permeabilities to both CO2 and HCO3-, we found that DIDS inhibited both Pm,HCO-3 and Pm,CO2, whereas intracellular CA activity remained unchanged. The mechanism by which DIDS reduces CO2 permeability may not be through an action on the lipid bilayer itself, but rather on a membrane transport protein, implying that this is a normal route for at least part of red cell CO2 exchange. PMID- 9861054 TI - Inositol 1,4,5-trisphosphate [correction of tris-phosphate] activation of inositol trisphosphate [correction of tris-phosphate] receptor Ca2+ channel by ligand tuning of Ca2+ inhibition. AB - Inositol 1,4,5-trisphosphate (IP3) [corrected] binding to its receptors (IP3R) in the endoplasmic reticulum (ER) activates Ca2+ release from the ER lumen to the cytoplasm, generating complex cytoplasmic Ca2+ concentration signals including temporal oscillations and propagating waves. IP3-mediated Ca2+ release is also controlled by cytoplasmic Ca2+ concentration with both positive and negative feedback. Single-channel properties of the IP3R in its native ER membrane were investigated by patch clamp electrophysiology of isolated Xenopus oocyte nuclei to determine the dependencies of IP3R on cytoplasmic Ca2+ and IP3 concentrations under rigorously defined conditions. Instead of the expected narrow bell-shaped cytoplasmic free Ca2+ concentration ([Ca2+]i) response centered at approximately 300 nM-1 microM, the open probability remained elevated (approximately 0.8) in the presence of saturating levels (10 microM) of IP3, even as [Ca2+]i was raised to high concentrations, displaying two distinct types of functional Ca2+ binding sites: activating sites with half-maximal activating [Ca2+]i (Kact) of 210 nM and Hill coefficient (Hact) approximately 2; and inhibitory sites with half-maximal inhibitory [Ca2+]i (Kinh) of 54 microM and Hill coefficient (Hinh) approximately 4. Lowering IP3 concentration was without effect on Ca2+ activation parameters or Hinh, but decreased Kinh with a functional half-maximal activating IP3 concentration (KIP3) of 50 nM and Hill coefficient (HIP3) of 4 for IP3. These results demonstrate that Ca2+ is a true receptor agonist, whereas the sole function of IP3 is to relieve Ca2+ inhibition of IP3R. Allosteric tuning of Ca2+ inhibition by IP3 enables the individual IP3R Ca2+ channel to respond in a graded fashion, which has implications for localized and global cytoplasmic Ca2+ concentration signaling and quantal Ca2+ release. PMID- 9861055 TI - A phytochrome from the fern Adiantum with features of the putative photoreceptor NPH1. AB - In plant photomorphogenesis, it is well accepted that the perception of red/far red and blue light is mediated by distinct photoreceptor families, i.e., the phytochromes and blue-light photoreceptors, respectively. Here we describe the discovery of a photoreceptor gene from the fern Adiantum that encodes a protein with features of both phytochrome and NPH1, the putative blue-light receptor for second-positive phototropism in seed plants. The fusion of a functional photosensory domain of phytochrome with a nearly full-length NPH1 homolog suggests that this polypeptide could mediate both red/far-red and blue-light responses in Adiantum normally ascribed to distinct photoreceptors. PMID- 9861056 TI - Origin of a chloroplast protein importer. AB - During evolution, chloroplasts have relinquished the majority of their genes to the nucleus. The products of transferred genes are imported into the organelle with the help of an import machinery that is distributed across the inner and outer plastid membranes. The evolutionary origin of this machinery is puzzling because, in the putative predecessors, the cyanobacteria, the outer two membranes, the plasma membrane, and the lipopolysaccharide layer lack a functionally similar protein import system. A 75-kDa protein-conducting channel in the outer envelope of pea chloroplasts, Toc75, shares approximately 22% amino acid identity to a similarly sized protein, designated SynToc75, encoded in the Synechocystis PCC6803 genome. Here we show that SynToc75 is located in the outer membrane (lipopolysaccharide layer) of Synechocystis PCC6803 and that SynToc75 forms a voltage-gated, high conductance channel with a high affinity for polyamines and peptides in reconstituted liposomes. These findings suggest that a component of the chloroplast protein import system, Toc75, was recruited from a preexisting channel-forming protein of the cyanobacterial outer membrane. Furthermore, the presence of a protein in the chloroplastic outer envelope homologous to a cyanobacterial protein provides support for the prokaryotic nature of this chloroplastic membrane. PMID- 9861057 TI - Abscisic acid-induced stomatal closure mediated by cyclic ADP-ribose. AB - Abscisic acid (ABA) is a plant hormone involved in the response of plants to reduced water availability. Reduction of guard cell turgor by ABA diminishes the aperture of the stomatal pore and thereby contributes to the ability of the plant to conserve water during periods of drought. Previous work has demonstrated that cytosolic Ca2+ is involved in the signal transduction pathway that mediates the reduction in guard cell turgor elicited by ABA. Here we report that ABA uses a Ca2+-mobilization pathway that involves cyclic adenosine 5'-diphosphoribose (cADPR). Microinjection of cADPR into guard cells caused reductions in turgor that were preceded by increases in the concentration of free Ca2+ in the cytosol. Patch clamp measurements of isolated guard cell vacuoles revealed the presence of a cADPR-elicited Ca2+-selective current that was inhibited at cytosolic Ca2+ >/= 600 nM. Furthermore, microinjection of the cADPR antagonist 8-NH2-cADPR caused a reduction in the rate of turgor loss in response to ABA in 54% of cells tested, and nicotinamide, an antagonist of cADPR production, elicited a dose-dependent block of ABA-induced stomatal closure. Our data provide definitive evidence for a physiological role for cADPR and illustrate one mechanism of stimulus-specific Ca2+ mobilization in higher plants. Taken together with other recent data [Wu, Y., Kuzma, J., Marechal, E., Graeff, R., Lee, H. C., Foster, R. & Chua, N.-H. (1997) Science 278, 2126-2130], these results establish cADPR as a key player in ABA signal transduction pathways in plants. PMID- 9861058 TI - Independent deletions of a pathogen-resistance gene in Brassica and Arabidopsis. AB - Plant disease resistance (R) genes confer race-specific resistance to pathogens and are genetically defined on the basis of intra-specific functional polymorphism. Little is known about the evolutionary mechanisms that generate this polymorphism. Most R loci examined to date contain alternate alleles and/or linked homologs even in disease-susceptible plant genotypes. In contrast, the resistance to Pseudomonas syringae pathovar maculicola (RPM1) bacterial resistance gene is completely absent (rpm1-null) in 5/5 Arabidopsis thaliana accessions that lack RPM1 function. The rpm1-null locus contains a 98-bp segment of unknown origin in place of the RPM1 gene. We undertook comparative mapping of RPM1 and flanking genes in Brassica napus to determine the ancestral state of the RPM1 locus. We cloned two B. napus RPM1 homologs encoding hypothetical proteins with approximately 81% amino acid identity to Arabidopsis RPM1. Collinearity of genes flanking RPM1 is conserved between B. napus and Arabidopsis. Surprisingly, we found four additional B. napus loci in which the flanking marker synteny is maintained but RPM1 is absent. These B. napus rpm1-null loci have no detectable nucleotide similarity to the Arabidopsis rpm1-null allele. We conclude that RPM1 evolved before the divergence of the Brassicaceae and has been deleted independently in the Brassica and Arabidopsis lineages. These results suggest that functional polymorphism at R gene loci can arise from gene deletions. PMID- 9861059 TI - The Arabidopsis thaliana RPM1 disease resistance gene product is a peripheral plasma membrane protein that is degraded coincident with the hypersensitive response. AB - Disease resistance in plants is often controlled by a gene-for-gene mechanism in which avirulence (avr) gene products encoded by pathogens are specifically recognized, either directly or indirectly, by plant disease resistance (R) gene products. Members of the NBS-LRR class of R genes encode proteins containing a putative nucleotide binding site (NBS) and carboxyl-terminal leucine-rich repeats (LRRs). Generally, NBS-LRR proteins do not contain predicted transmembrane segments or signal peptides, suggesting they are soluble cytoplasmic proteins. RPM1 is an NBS-LRR protein from Arabidopsis thaliana that confers resistance to Pseudomonas syringae expressing either avrRpm1 or avrB. RPM1 protein was localized by using an epitope tag. In contrast to previous suggestions, RPM1 is a peripheral membrane protein that likely resides on the cytoplasmic face of the plasma membrane. Furthermore, RPM1 is degraded coincident with the onset of the hypersensitive response, suggesting a negative feedback loop controlling the extent of cell death and overall resistance response at the site of infection. PMID- 9861061 TI - Brain-wave recognition of sentences. AB - Electrical and magnetic brain waves of two subjects were recorded for the purpose of recognizing which one of 12 sentences or seven words auditorily presented was processed. The analysis consisted of averaging over trials to create prototypes and test samples, to each of which a Fourier transform was applied, followed by filtering and an inverse transformation to the time domain. The filters used were optimal predictive filters, selected for each subject. A still further improvement was obtained by taking differences between recordings of two electrodes to obtain bipolar pairs that then were used for the same analysis. Recognition rates, based on a least-squares criterion, varied, but the best were above 90%. The first words of prototypes of sentences also were cut and pasted to test, at least partially, the invariance of a word's brain wave in different sentence contexts. The best result was above 80% correct recognition. Test samples made up only of individual trials also were analyzed. The best result was 134 correct of 288 (47%), which is promising, given that the expected recognition number by chance is just 24 (or 8.3%). The work reported in this paper extends our earlier work on brain-wave recognition of words only. The recognition rates reported here further strengthen the case that recordings of electric brain waves of words or sentences, together with extensive mathematical and statistical analysis, can be the basis of new developments in our understanding of brain processing of language. PMID- 9861060 TI - Verb generation in patients with focal frontal lesions: a neuropsychological test of neuroimaging findings. AB - What are the neural bases of semantic memory? Traditional beliefs that the temporal lobes subserve the retrieval of semantic knowledge, arising from lesion studies, have been recently called into question by functional neuroimaging studies finding correlations between semantic retrieval and activity in left prefrontal cortex. Has neuroimaging taught us something new about the neural bases of cognition that older methods could not reveal or has it merely identified brain activity that is correlated with but not causally related to the process of semantic retrieval? We examined the ability of patients with focal frontal lesions to perform a task commonly used in neuroimaging experiments, the generation of semantically appropriate action words for concrete nouns, and found evidence of the necessity of the left inferior frontal gyrus for certain components of the verb generation task. Notably, these components did not include semantic retrieval per se. PMID- 9861062 TI - Introduction: pediatric cardiology exercise testing. PMID- 9861064 TI - Comments PMID- 9861063 TI - Normative cardiovascular responses to exercise in children. PMID- 9861065 TI - Cardiorespiratory testing: anaerobic threshold/respiratory threshold. PMID- 9861066 TI - Comments PMID- 9861068 TI - Comments PMID- 9861067 TI - Velocity of oxygen uptake response at the onset of exercise: A comparison between children after cardiac surgery and healthy boys. AB - A comparison was carried out concerning maximal oxygen uptake, oxygen uptake adjustment at the onset of high-intensity exercise, and maximal blood lactate between 10 healthy prepubertal boys and 35 children after repair of cardiac malformations or after Fontan operation. Mean maximal oxygen uptake (VO2) was moderately reduced in children after repair of tetralogy of Fallot or after Mustard or Senning operations and severely reduced after Fontan operations. Conversely, mean half-time of VO2 response was moderately prolonged in children after repair of tetralogy of Fallot or after Senning and Mustard operations and considerably prolonged after Fontan operations. According to our results unfavorable kinetics of VO2 response to physical exercise are present in addition to reduced aerobic power in many of the operated children. Besides being less qualified for endurance performance, these children are also less prepared for short, high-intensity exercise. PMID- 9861069 TI - Pre- and postoperative exercise testing of the child with atrial septal defect. PMID- 9861070 TI - Comments PMID- 9861072 TI - Comments PMID- 9861071 TI - Exercise testing in children with pulmonary valvar stenosis. AB - Pulmonary valvar stenosis with intact ventricular septum is a common anomaly. This lesion poses a fixed obstruction to the right ventricular outflow. The right ventricle ejects the entire cardiac output across the stenotic valve. Right ventricular systolic pressure and oxygen demand are increased at rest and more so with exercise. Exercise tolerance in children and adults with mild valvar pulmonary stenosis is nearly normal, but is diminished in those with moderate and severe stenosis, indicating impaired ability to sustain adequate cardiac output. Following relief of stenosis, cardiac performance improves in children, but remains abnormal in adults. This appears to be related to postoperative resolution of right ventricular hypertrophy in children, whereas myocardial fibrosis may explain the lack of improvement in adults. PMID- 9861074 TI - Comments PMID- 9861073 TI - Pre- and postoperative exercise testing of the child with coarctation of the aorta. AB - Exercise studies in patients with coarctation of the aorta (CoA) are reviewed. A brief survey of the natural history of unoperated CoA, the pioneers who reported the first operative repair, and comments on blood pressure measurements are provided. Preoperative and postoperative (postop) exercise studies of patients with CoA are also reviewed. The various etiological factors which may contribute to hypertension found in some post-op CoA patients are discussed. PMID- 9861075 TI - Exercise studies in tetralogy of Fallot: a review. PMID- 9861076 TI - Comments PMID- 9861077 TI - Exercise studies in patients with transposition of the great arteries after atrial repair operations (Mustard/Senning): a review. AB - Exercise evaluation studies of patients after atrial repair surgery for transposition of the great arteries, as in tetralogy of Fallot, represent only a small fraction of the 3970 Medline references (1966 to mid-1997) concerning this congenital heart lesion. We have abstracted data from 27 studies from 20 institutions reporting on measurements during exercise on work capacity, heart rate response, respiratory gas exchange, or radionuclide/radiographic systemic ventricular ejection fraction measurements in addition to resting pulmonary function measurements. These studies provide almost uniform general conclusions that even after 20 or more years of follow-up (1) most patients "report" that they are asymptomatic in performing usual levels of physical activities; (2) significant abnormalities are present, often in more than half of the patients studied, in one or more of the exercise measurements when compared to control subjects; and (3) the diminished exercise performance is related to a diminished cardiac output, results from diminished stroke volume but is also related to a blunted heart rate response. PMID- 9861078 TI - Comments PMID- 9861080 TI - Comments PMID- 9861079 TI - Exercise testing after the Fontan operation. PMID- 9861081 TI - Exercise testing in children with primary pulmonary hypertension. AB - Cardiopulmonary exercise testing is a useful noninvasive tool to assess physiological changes associated with exercise. Developing noninvasive methods to assess the severity of cardiopulmonary disorders, as well as the response to therapeutic interventions, is useful in conditions, such as primary pulmonary hypertension, in which invasive procedures carry significant risks. The 6-minute walk test is a simple measure of exercise endurance. Exercise studies that measure both hemodynamic and ventilatory responses provide additional information regarding the interaction of the circulatory and pulmonary systems. Subtle changes in exercise capacity may suggest deterioration prior to clinical manifestations. This may lead to an earlier reevaluation, including repeat cardiac catheterization, and subsequently changes in medical and/or surgical therapy. PMID- 9861082 TI - Comments PMID- 9861083 TI - Exercise in hypertensive children and adolescents: any harm done? AB - Hypertensive children and adolescents are frequently arbitrarily excluded from sports or exercise because of the fear of possible complications, such as stroke or myocardial infarction. No hypertensive children have had exercise-related morbidity or mortality in the reviewed literature. No restriction from dynamic exercise seems warranted; training has led to reductions in blood pressure in hypertensive youth. Isometric exercise is more hotly debated. Because no complications of exercise have been reported, the author prefers to allow participation if no target organ damage is present. Life-long maintenance of habitual physical activity may help prevent adult-onset essential hypertension. PMID- 9861084 TI - Comments PMID- 9861085 TI - Exercise and lipid abnormalities. PMID- 9861086 TI - Comments PMID- 9861088 TI - Comments PMID- 9861087 TI - Exercise testing of the child with obesity. AB - As the prevalence of childhood obesity increases, exercise testing of obese children is likely to increase as well. This article discusses the implications of pediatric obesity for exercise testing and provides some recommendations for conducting tests and evaluating results. Studies comparing obese and nonobese children during exercise testing indicate that obese children are capable of meeting the challenges of exercise testing to nearly the same extent as their nonobese peers. Their physiologic responses, at least for the levels of obesity reported in the literature, are not sufficiently different from their nonobese counterparts to necessitate major changes in test protocols. Laboratory staff should pay special attention to fostering confidence in the obese child during the pretest routine. PMID- 9861090 TI - Comments PMID- 9861089 TI - Exercise testing as a rehabilitative/training tool. AB - Exercise testing is an exceedingly useful noninvasive method for assessing cardiovascular function not only at rest but also during programmed, supervised physical exercise. Exercise testing has been intensively studied to delineate the cardiovascular response in various disorders [3-5, 15, 19, 22, 23, 26, 32]. Exercise testing as a tool for rehabilitation and for training to perform physical activity is discussed here. PMID- 9861092 TI - From other journals PMID- 9861093 TI - Upcoming events in pediatric cardiology PMID- 9861091 TI - Conclusion: pediatric cardiology exercise testing PMID- 9861094 TI - Editorial PMID- 9861095 TI - Hemodialysis prognostic nutrition index as a predictor for morbidity and mortality in hemodialysis patients and its correlation to adequacy of dialysis. Council on Renal Nutrition National Research Question Collaborative Study Group. AB - OBJECTIVE: Prospectively examine the use of a hemodialysis prognostic nutrition index (HD-PNI) as a predictor for morbidity and mortality in hemodialysis patients and its correlation to adequacy of dialysis. DESIGN: Prospective randomized collaborative study group. SETTING: There were 211 chronic hemodialysis centers; 202 from 43 United States, 9 from Canada. PATIENTS: There were 1527 hemodialysis patients undergoing treatment a minimum of 3 months and at least 18 years of age. Sample mirrored United States Renal Data System data for age, sex, race, and etiology of renal failure. INTERVENTIONS: None; routinely collected demographic, biochemical, and clinical data for 8-month baseline and 3 month predictive phases. METHODS: HD-PNI calculated from baseline data as linear mathematical equation using level of serum albumin, level of serum creatinine, and number of days and times hospitalized; HD-PNI risk defined as >/=0.8. Adequacy of dialysis calculated as urea reduction ratio (URR) from baseline data; adequacy risk defined as URR of /=65% did not significantly improve prediction. CONCLUSIONS: Use of HD-PNI is an effective screening tool to identify hemodialysis patients at risk for morbidity and mortality. No correlation was found between URR and HD-PNI. PMID- 9861096 TI - Quality of life implications of inadequate protein nutrition among hemodialysis patients. AB - OBJECTIVE: To determine the quality of life implications of inadequate protein nutrition among hemodialysis patients. DESIGN: Cross-sectional study. SETTING: All 22 chronic hemodialysis units in northeast Ohio. PATIENTS: There were 289 randomly selected patients. INTERVENTION: Interview and chart abstraction. MAIN OUTCOME MEASURES: Six quality of life subscales related to general health, physical function, social function, symptoms, burden of kidney disease, and employment status. RESULTS: Low albumin levels were independently associated with decreased physical function, social function, and burden of kidney disease scores after adjustment for patient age, race, gender, health insurance, cause of renal failure, years on dialysis, and number of comorbid conditions. Low protein catabolic rate was independently associated with decreased physical function scores and with being retired or disabled. CONCLUSION: Inadequate protein nutrition is independently associated with poor quality of life. Further study is needed to develop interventions to overcome barriers to protein nutrition and to determine the effect of such interventions on protein nutrition and patient quality of life. PMID- 9861097 TI - Reported pica behavior in a sample of incident dialysis patients. AB - In a prospective study, pica behavior was investigated during baseline interviews with a cohort of incident patients (n = 226) who began chronic dialysis therapy in metropolitan Atlanta, GA, during 1996 to 1997. Pica, defined as current pica behavior and/or reported history of pica behavior, was reported by 16% of the sample. Patients reporting pica were significantly more likely to be African American women and were significantly younger than the remainder of the sample. Approximately two thirds of patients who reported pica behaviors craved and excessively consumed ice; the remainder craved and consumed starch, dirt, flour, or aspirin. Among patients reporting pica, average serum albumin values were low and average phosphorus was increased. The average hematocrit of patients reporting ice pica was low. Over half of the hemodialysis patients reporting pica behavior had excessive usual interdialytic weight gain. Potential symptoms/problems affecting quality of life among patients practicing pica, eg, cramps, are shown in a case report. The data indicate the need for targeted education and support for dietitians' increased interaction with dialysis patients involved in pica behaviors. PMID- 9861098 TI - A dietary survey in Indian hemodialysis patients. AB - Malnutrition is a common problem in maintenance hemodialysis (MHD) patients, and compromised intake is an important cause. There is no information available about the nutrient intakes of MHD patients in India. The nutrient intakes of 106 MHD patients were studied cross-sectionally and on follow-up. A 24-hour recall was used on 4 consecutive days. After 2 months on dialysis, the mean energy intake was 29 +/- 6.6 kcal/kg ideal body weight (IBW) and the mean protein intake was.93 +/-.39 g/kg IBW (high biological value [HBV] protein 49% +/- 8.5%). Dietary deficiency of both protein and calories was present in 64.9%. Intake was better on nondialysis days compared with dialysis days, and in women and older patients. On follow-up there was no significant increase in food intake up to 6 months. After that, the total calorie intake increased significantly with a disproportionate drop in high biological value protein consumed and appeared to be derived predominantly from carbohydrate food (mean kcal/kg, 37 +/- 6.9; mean protein g/kg, 0.96 +/- 0.19; ratio of HBV protein to total protein consumed,.42 +/-.09). In summary this study showed suboptimal energy and protein intake in an MHD population. Intakes were further compromised on dialysis days, and with increasing time spent on dialysis, the quality of nutrient intake became poorer. PMID- 9861100 TI - Response to intradialytic parenteral nutrition. AB - Malnutrition is a major clinical problem in patients receiving maintenance hemodialysis and has adverse effects on survival. Nutritional intervention is indicated, and there is evidence that intradialytic parenteral nutrition can be beneficial. We describe the application of a formal policy regarding the use of intradialytic parenteral nutrition and the beneficial effects on nutrition in the first four patients managed in this fashion. However, the fifth patient did not respond to parenteral nutrition, despite adequate dialysis. This prompted further investigation, and the patient was shown to have extensive gastric malignancy. This report shows that establishing a protocol for intradialytic parenteral nutrition is possible in a medium-sized hemodialysis unit. In these circumstances, nonresponse to this intervention should always be investigated to determine if there is another underlying cause of malnutrition unrelated to renal failure. PMID- 9861099 TI - A comparison of two methods of dietary assessment in peritoneal dialysis patients. AB - OBJECTIVE: To conduct a comparison of two methods of dietary assessment in patients on peritoneal dialysis. DESIGN: Comparative, cross-sectional study of two methods of dietary assessment (3-day diet diary and 24-hour recall). Data was collected simultaneously by a single experienced dietitian. Each assessment was coded and analyzed blind. SETTING: Regional speciality peritoneal dialysis training unit. PATIENTS: In this study, 30 peritoneal dialysis patients recruited prospectively and consecutively as they attended for out-patient assessment of dialysis adequacy. Age range was 22 to 77 years. Patients were excluded if unwell, younger than 18 years, or had peritonitis. OUTCOME MEASURES: Total energy and protein intakes from both methods were compared. Protein intakes from both methods were compared with the protein catabolic rate generated from urea kinetics. Data obtained from both methods were compared using paired t tests, linear regression, and Bland and Altman techniques. RESULTS: There were no significant differences in the mean daily protein (72.4 g v 76.6 g) and total energy (1757 kcals (7.35 MJ) v. 1897 kcals (7.94 MJ)) intakes determined by the two methods. Positive correlations were seen between the measurements for protein intake (r =.58, P =.0026) energy intake (r =.78, P <.00001), with mean differences of.066 g/kg/d (SD.38) 2.04 kcal/kg/day (SD 6.67), respectively. For both methods there was a similar positive correlation between dietary protein intake and protein catabolic rate. A 24-hour recall was more likely to result in successful collection of data (29 of 30) than 3-day diet diaries (25 of 30) and was less time consuming. CONCLUSION: These two methods of determining dietary protein and energy intake do not differ significantly in the information they provide. The relative success in obtaining completed records of intake, the shorter time taken and the opportunity for patient education and assessment of other nutrition related factors has led to the adoption of the 24-hour recall method in our institution. PMID- 9861103 TI - Literature review. PMID- 9861102 TI - Initial nutrition assessment. PMID- 9861101 TI - Treating fluid noncompliance in the hemodialysis population using unit wide contests. AB - Our dialysis unit observed fluid noncompliance over a wide variety of patients. The consequences of fluid abuse that can include systemic and cardiovascular overload are a frequent clinic complication of hemodialysis patients. In spite of all our attempts, our dialysis unit continued to have a substantial group of patients who could not adhere to their fluid restriction. Our dialysis unit needed a novel program to help motivate patients to comply with diet/medication fluid regimens. A method that seems to work well in our dialysis unit is playing unit-wide games. We developed "The Fluid Game," an original patient education idea, for providing renal patients with a fun incentive to keep their interdialytic weight gains within acceptable limits. PMID- 9861104 TI - How to get the most from your iron supplement. PMID- 9861106 TI - Message from the chairperson PMID- 9861105 TI - Vending machine snacks. PMID- 9861107 TI - Announcements PMID- 9861108 TI - Education and training in airway management. PMID- 9861109 TI - Adrenocortical function in critical illness. PMID- 9861110 TI - Effect of graft reperfusion on haemodynamics and gas exchange during liver transplantation. AB - We have documented the changes in gas exchange, haemodynamic state and associated physiological variables which occurred after graft reperfusion in 20 patients undergoing uncomplicated orthotopic liver transplantation. Gas exchange was measured during constant ventilation using a metabolic monitor. After reperfusion, there were increases in VO2 (mean increase 57 (SD 25) ml min-1) (P < 0.001), VCO2 (mean increase 38 (17) ml min-1) (P < 0.001) and PaCO2 (mean increase 0.88 (0.56) kPa) (P < 0.001). These were associated with increases in cardiac output (1.2 (1.0) litre min-1 m-2) (P < 0.001) and mean pulmonary artery pressure (9 (6) mm Hg) (P < 0.001). There was a decrease in standard bicarbonate concentration (0.96 (1.6) mmol litre-1) (P < 0.02) and increase in hydrogen ion concentration (8.15 (5.9) mmol litre-1) (P < 0.001) consistent with the release of an acid load from the graft and previously ischaemic tissues. The increases in PaCO2 and hydrogen ion concentration were significantly larger in patients in whom venovenous bypass was used during the anhepatic period compared with the "piggyback" surgical technique. We found correlations between the changes in PaCO2 and VCO2 (r2 = 0.25, P < 0.02), cardiac output and VCO2 (r2 = 0.34, P < 0.01), and cardiac output and VO2 (r2 = 0.34, P < 0.01). We conclude that major alterations in gas exchange occur after reperfusion which result from alterations in metabolic rate and haemodynamic changes. These may be clinically relevant, particularly in patients at risk of cerebral oedema. PMID- 9861111 TI - Factors affecting assessment of cerebral autoregulation using the transient hyperaemic response test. AB - The transient hyperaemic response in the middle cerebral artery blood flow velocity on the release of brief compression of the ipsilateral common carotid artery has been validated as an indicator of cerebral autoregulation. We evaluated, in three stages, the effect of experimental factors such as duration of compression of the common carotid artery and magnitude of the decrease in blood flow velocity during common carotid artery compression on the transient hyperaemic response. In stage 1, 13 healthy volunteers underwent six transient hyperaemic response tests each; two tests each for either 3, 6 or 10 s duration of compression of the common carotid artery. In stage 2, 10 volunteers underwent four transient hyperaemic response tests each; two tests each for either 10 or 15 s duration of compression of the common carotid artery. In stage 3, data from the transient hyperaemic response tests using 10 s compression from the 23 volunteers who participated in stages 1 and 2 were analysed to evaluate the relationship between magnitude of decrease in blood flow velocity at the onset of compression and the transient hyperaemic response. The transient hyperaemic response ratio (blood flow velocity after the release of compression/baseline blood flow velocity) increased significantly when the duration of common carotid artery compression increased from 3 to 6 s, or from 6 to 10 s (stage 1); increase in the duration from 10 to 15 s did not have any significant effect (stage 2). The transient hyperaemic response ratio correlated significantly with the magnitude of decrease in blood flow velocity after compression, up to the values of the compression ratio (percent decrease in blood flow velocity at the onset of compression) of 40% but not more (stage 3). We conclude that experimental factors such as duration of common carotid artery compression and magnitude of the decrease in blood flow velocity during common carotid artery compression can significantly influence the transient hyperaemic response. These factors should be controlled if the transient hyperaemic response test is used for a comparison between repeated measurements. A compression time of 10 s and a compression ratio of 40% or more, allow maximum expression of the hyperaemic response in healthy volunteers. PMID- 9861112 TI - Influence of dose of domperidone on the acute ventilatory response to hypoxia in humans. AB - We have studied the ventilatory responses to acute isocapnic hypoxia (SpO2 78.8 (SD 1.4)% for 10 min) in 10 male volunteers given three different doses of oral domperidone: placebo, domperidone tablets 10 mg, 20 mg or 30 mg every 8 h for 48 h on separate days. Neither baseline ventilation nor the acute hypoxic ventilatory response was significantly different from placebo for any of the domperidone doses. However, hypoxic responses were either increased with increments of domperidone or subjects were not sensitive. We arbitrarily divided subjects into two groups according to their hypoxic response-plasma domperidone concentration relationship. Analysis of subjects (n = 5) who demonstrated at least a 2-litre min-1 increase in ventilation per 10 ng ml-1 increase in plasma domperidone concentration showed the greatest augmentation of hypoxic responses with the 20-mg dose (median 19.45 (range 13.37, 22.30) litre min-1) compared with placebo (median 8.21 (3.74, 9.47)) (P = 0.003). We were unable to predict which subjects would be sensitive to the effects of domperidone. PMID- 9861113 TI - A physiology simulator: validation of its respiratory components and its ability to predict the patient's response to changes in mechanical ventilation. AB - We aimed to validate the mathematical validity and accuracy of the respiratory components of the Nottingham Physiology Simulator (NPS), a computer simulation of physiological models. Subsequently, we aimed to assess the accuracy of the NPS in predicting the effects of a change in mechanical ventilation on patient arterial blood-gas tensions. The NPS was supplied with the following measured or calculated values from patients receiving intensive therapy: pulmonary shunt and physiological deadspace fractions, oxygen consumption, respiratory quotient, cardiac output, inspired oxygen fraction, expired minute volume, haemoglobin concentration, temperature and arterial base excess. Values calculated by the NPS for arterial oxygen tension and saturation (PaO2 and SaO2), mixed venous oxygen tension and saturation (PvO2 and SvO2), arterial and mixed venous carbon dioxide tension (PaCO2 and PvCO2) and arterial pH were accurate compared with measured values. Subsequently, arterial gas responses to changes in minute volume of FiO2 were measured in 31 patients and were compared with the NPS prediction for each response. The 95% limits of agreement in predicting the magnitude of change were: arterial oxygen tension -2.07 to 2.47 kPa; PaCO2 -0.33 to 0.67 kPa; and pH -0.023 to 0.033. This investigation has validated respiratory components of the NPS. We recommend the NPS as a clinical tool for predicting the effects of alterations in mechanical ventilation in stable patients in the intensive care unit. PMID- 9861114 TI - Respiratory sinus arrhythmia and clinical signs of anaesthesia in children. AB - We have investigated changes in respiratory sinus arrhythmia (RSA) and compared these with clinical signs of anaesthesia in children. Children aged 3-10 yr were anaesthetized by gaseous induction with halothane and nitrous oxide. Multiple heart rate variability (HRV) spectra were obtained by power spectral analysis of continuous epochs of time from before introduction of halothane (baseline) until the pupils were central and fixed (stage 3). Measurement of RSA was performed by integration of the area under the spectral curve within the range of the respiratory frequency +/- 0.15 Hz. In all patients RSA decreased continuously during induction unless stimulation occurred with insertion of an airway. Values of RSA were compared at three times: baseline, loss of pharyngeal tone and stage 3. The decrease in RSA from baseline to loss of pharyngeal tone and from loss of pharyngeal tone to stage 3 was significant (P = 0.003 and P = 0.018, respectively). These results show that RSA can be related to the clinical signs of anaesthesia and has potential as a measure of depth of anaesthesia in children. PMID- 9861115 TI - Changes in blood-gas tensions during apnoeic oxygenation in paediatric patients. AB - We report changes in arterial blood-gas tensions for up to 5 min of apnoeic oxygenation in 26 anaesthetized paediatric patients (21 children, five infants). Changes in oxygen and carbon dioxide tension were greatest in the first minute of apnoeic oxygenation. In subsequent minutes, rates of change in gas tension were approximately constant. The rate of decline in oxygen tension (31 (95% confidence interval (CI) 20.1-42.2) mm Hg min-1) was more than three times that reported in studies in adults. The rate of increase in carbon dioxide tension (4.2 (95% CI 3.7-4.7) mm Hg min-1) was similar to that reported in adults. After successful preoxygenation, oxygen tension remained greater than 290 mm Hg in all children (age > 1 yr) throughout the study. This was not the case in infants. We found no correlation between changes in blood-gas tensions and age or weight of patients. The small number of infants studied showed rapid decreases in oxygen tension which if sustained would be expected to limit the safe duration of apnoeic oxygenation, unlike adults where apnoeic oxygenation is limited by hypercapnia. Extrapolation of our results suggests that when preoxygenation has been successful, apnoeic oxygenation could continue safely in children for at least 10 min. Infants may become hypoxic after only 2 min. PMID- 9861116 TI - Beta 2-adrenergic responsiveness in vivo during abdominal surgery. AB - We have studied adrenergic function in vivo during anaesthesia and surgery. Epinephrine 50 ng kg-1 min-1 was given by i.v. infusion over 30 min to 10 healthy adult volunteers and to 10 patients undergoing abdominal operations. The cAMP response to stimulation by epinephrine, which was obtained as the area under the curve (AUC) for plasma cAMP concentration divided by the AUC for plasma concentration of epinephrine, was more pronounced during surgery (mean ratio 3.5) than in the control situation (ratio 1.4; P < 0.02). This resulted in greater hypokalaemic and hyperglycaemic responses (ratios -0.67 and 4.5) than in the control group (ratios -0.33 and 1.6, respectively; P < 0.004). Mean arterial pressure decreased in the control group while it increased in the study group, and serum cortisol concentration was higher in those who underwent surgery (P < 0.02). These results are consistent with an increased adrenergic response during abdominal surgery. PMID- 9861117 TI - Effect of preoperative extradural bupivacaine and morphine on stump sensation in lower limb amputees. AB - We have examined the effect of preoperative extradural bupivacaine and morphine on postoperative stump sensation in 31 patients undergoing amputation of the lower limb in a prospective, randomized, double-blind study. Patients were allocated randomly to one of two groups: group 1 received extradural 0.25% bupivacaine 4-7 ml h-1 and morphine 0.16-0.28 ml h-1 before and during operation; group 2 received extradural saline before and during amputation and conventional analgesics for pain treatment. All patients received general anaesthesia for the amputation and extradural bupivacaine and morphine after operation. Sensory examination of the limb/stump was carried out before amputation, and after 1 week and 6 months. The following were measured: pressure pain thresholds (pressure algometry), touch and pain detection thresholds (von Frey hairs), thermal sensibility (thermal rolls), and allodynia and wind-up-like pain. There were no differences between the two groups at any of the postoperative assessments for mechanical and thermal sensibility or rate of allodynia and wind-up-like pain. Our study suggests that preoperative and intraoperative extradural block had no long-term prophylactic effect on hyperalgesia, allodynia or wind-up-like pain. PMID- 9861118 TI - Does speed of intrathecal injection affect the distribution of 0.5% hyperbaric bupivacaine? AB - We have evaluated the influence of speed of intrathecal injection on lateral distribution of 0.5% hyperbaric bupivacaine. We studied 60 patients undergoing lower limb surgery who were placed in the lateral position with the operative side in the dependent position. After dural puncture (25-gauge Whitacre spinal needle), the needle aperture was turned towards the dependent side and 0.5% hyperbaric bupivacaine 8 mg was injected randomly at a rate of 0.02 ml s-1 (group slow, n = 30) or 0.25 ml s-1 (group fast, n = 30). Lateral position was maintained for 15 min while a blinded observer recorded loss of pinprick sensation and degree of motor block on both surgical and non-surgical sides. There were no differences between the groups. Forty-five minutes after patients were turned to the supine position, spinal anaesthesia was unilateral in 17 patients in group slow (56%) and in 13 patients in group fast (43%). We conclude that using extremely low speeds for intrathecal injection were not clinically advantageous in obtaining unilateral spinal anaesthesia. PMID- 9861119 TI - Effect of nimodipine on regression of spinal analgesia. AB - We have investigated the effect of infusion of nimodipine on the spread of spinal anaesthesia in 50 patients undergoing transurethral procedures. Patients were allocated randomly to receive during operation continuous infusion of nimodipine 10 ml h-1 (group N, n = 25) or normal saline (group C, n = 25) in a double-blind manner. All patients received hyperbaric lidocaine 100 mg (5% in 8% dextrose) intrathecally and were then placed in the lithotomy position. Twenty minutes after intrathecal injection the level of spinal anaesthesia was tested with a pressure palpator and a baseline was established. Assessments were repeated 5, 10 and 15 min thereafter. Five minutes after establishing baseline, mean regression of sensory analgesia did not differ between groups. Analgesia had regressed by 1.3 (SD 1.4) and 1.0 (1.9) cm, respectively. After 10 min, sensory block in group N regressed by 1.7 (1.7) cm and in group C by 1.5 (1.6) cm. After 15 min these values were 1.1 (1.7) cm and 2.2 (1.9) cm, respectively (P < 0.035). Similar results were found after normalizing the changes by dividing the change by patient height. PMID- 9861120 TI - An open, randomized comparison of alfentanil, remifentanil and alfentanil followed by remifentanil in anaesthesia for craniotomy. AB - We studied 52 adults undergoing elective craniotomy, allocated randomly to one of three opioid treatments: alfentanil 50 micrograms kg-1 followed by 0.833 microgram kg-1 min-1 until dural closure (group Alf.); alfentanil 50 micrograms kg-1 followed by 0.833 microgram kg-1 min-1 for 2 h, then remifentanil 0.25 microgram kg-1 min-1 (group Alf.-Remi.); or remifentanil 1 microgram kg-1 followed by 0.5 microgram kg-1 min-1 reducing to 0.25 microgram kg-1 min-1 after craniotomy (group Remi.). Anaesthesia was maintained with infusion of propofol and 66% nitrous oxide in oxygen. Infusions of propofol and remifentanil were stopped at head bandaging. Group Remi. had the least intraoperative haemodynamic responses and group Alf. the most (P < 0.05). Times to tracheal extubation and obey commands were similar in all groups. In all patients in group Alf.-Remi. and group Remi., the trachea was extubated 27 min from the end of anaesthesia; three patients in group Alf. were slower to recover. Use of analgesia in the recovery room and time to transfer to the neurosurgical unit were similar in the three groups. PMID- 9861121 TI - Patient-maintained remifentanil target-controlled infusion for the transition to early postoperative analgesia. AB - We studied 30 male patients in the early postoperative period to assess the efficacy, safety and feasibility of a patient-demand, target-controlled infusion (TCI) of remifentanil. All patients received the same TCI-based propofol remifentanil anaesthetic for elective orthopaedic surgery. At the end of surgery, infusion of remifentanil was reduced progressively until patients were breathing spontaneously. After extubation and transfer to the post-anaesthesia care unit, patients were given control of a hand-set and were able to increase the target remifentanil blood concentration by increments of 0.2 ng ml-1. If there were no demands, the TCI controller automatically reduced the target concentration. Pain scores, sedation level, ventilatory frequency, oxygen saturation and nausea were assessed. Mean time to onset of satisfactory analgesia (VAS < or = 3, out of 10) was 18.9 (95% confidence interval (Cl) 15.8-21.9) min at a mean target remifentanil concentration of 2.02 (Cl 1.87-2.16) ng ml-1. There were no episodes of hypoxaemia and the lowest ventilatory frequency was 9 bpm. Nausea occurred in 26.6% of patients and 10% vomited. The majority of patients were only slightly sedated. These results imply an effective tool without respiratory side effects in the early postoperative period after anaesthesia using remifentanil as the analgesic component. PMID- 9861122 TI - Ketorolac, diclofenac and ketoprofen are equally efficacious for pain relief after total hip replacement surgery. AB - We have compared the efficacy of ketorolac 30 mg i.v. followed by infusion at a rate of 90 mg/15.5 h, with that of diclofenac 75 mg followed by infusion of 75 mg/15.5 h or ketoprofen 100 mg followed by infusion of 100 mg/15.5 h, on postoperative pain in 85 patients after hip replacement surgery under spinal anaesthesia in a prospective, double-blind, randomized study. Supplementary analgesia was administered during the 16-h postoperative period with bolus doses of fentanyl delivered by a patient-controlled analgesia system. Mean total consumption of PCA-administered fentanyl was 890 (SD 400) micrograms in the ketorolac group, 920 (550) micrograms in the diclofenac group and 850 (350) micrograms in the ketoprofen group (ns). Median VAS scores were low over the entire study in each group and there was no significant difference between groups. No serious adverse events were recorded. PMID- 9861123 TI - Influence of timing of morphine administration on postoperative pain and analgesic consumption. AB - We have investigated if a pre-emptive dose of morphine, given 30 min before skin incision, influenced postoperative pain and morphine consumption after hysterectomy. In a prospective, randomized, double-blind, placebo-controlled clinical study, patients received morphine 0.3 mg kg-1 at induction of anaesthesia or 30 min later at skin incision. The primary endpoint was defined as 24-h morphine consumption via patient-controlled analgesia. We could not demonstrate any difference between the two groups in morphine consumption or pain scores, and we conclude that there was no evidence of pre-emptive analgesia in this study. PMID- 9861124 TI - A qualitative systematic review of incisional local anaesthesia for postoperative pain relief after abdominal operations. AB - In a qualitative systematic review, we have evaluated randomized controlled trials (RCT) of incisional local anaesthesia compared with placebo or no treatment in the control of postoperative pain after open abdominal operations. Twenty-six studies with data from 1211 patients were considered appropriate for analysis. Five RCT considered inguinal herniotomy, four hysterectomy, eight cholecystectomy and nine studies a variety of surgical procedures. Outcome measures were pain scores, supplementary analgesics and time to first analgesic request. Efficacy was estimated by significant difference (P < 0.05), as reported in the original investigation. All studies of herniotomy showed a 2-7-h duration of clinically relevant improved pain relief. Results of hysterectomy studies were inconclusive, with two being negative. Five of the cholecystectomy studies showed significant differences but questionable clinical importance and validity in three. In various other procedures results were inconsistent and in some of minor clinical importance. Except for herniotomy, there was a lack of evidence for effect of incisional local anaesthesia on postoperative pain and further standardized studies are needed before recommendations can be made. PMID- 9861125 TI - High-dose ondansetron regimen vs droperidol for morphine patient-controlled analgesia. AB - We have performed a randomized, double-blind study comparing droperidol and high dose ondansetron mixed with morphine for patient-controlled analgesia (PCA). To detect a reduction in the incidence of postoperative nausea and vomiting from 55% to 20% with a power of 80% at the P < 0.05 level, 29 patients per group were required. We studied 60 healthy women undergoing abdominal hysterectomy, anaesthetized using a standard technique. Group D received a bolus dose of droperidol 1.25 mg at induction followed by droperidol 0.1 mg per 1 mg of morphine from the PCA system. Group O received a bolus dose of ondansetron 4 mg at induction followed by ondansetron 0.32 mg per 1 mg of morphine. This dose of ondansetron is more than double that studied previously. Mean nausea and vomiting scores at 4, 8, 12 and 24 h, mean time to first vomit, sedation scores, incidence of side effects, and doses of prochlorperazine did not differ between the groups. In group D, 24 patients did not vomit compared with 23 in group O. The only significant difference between the groups was increased morphine consumption in the ondansetron group up until 12 h after operation (P < 0.05), but by 24 h this difference was not significant. The ondansetron regimen was more expensive (at local prices) by a factor of 27, and our results suggested no clinical advantage over droperidol. PMID- 9861126 TI - Granisetron-droperidol combination for the prevention of postoperative nausea and vomiting in female patients undergoing breast surgery. AB - We have compared the efficacy and safety of the combination granisetron droperidol with each antiemetic alone in preventing postoperative nausea and vomiting (PONV) after breast surgery. In a randomized, double-blind study, 150 female patients received granisetron 3 mg, droperidol 1.25 mg or granisetron 3 mg with droperidol 1.25 mg (n = 50 each) i.v., immediately before induction of anaesthesia. A standard general anaesthetic technique was used. The incidence of PONV during the first 24 h after anaesthesia was 18% with granisetron, 38% with droperidol and 4% with the granisetron-droperidol combination (P < 0.05; overall Fisher's exact probability test). We conclude that the granisetron-droperidol combination was more effective than each antiemetic alone in the prevention of PONV in female patients undergoing breast surgery. PMID- 9861127 TI - Oral granisetron prevents postoperative vomiting in children. AB - We have studied the efficacy of granisetron, a selective 5-hydroxytryptamine type 3 receptor antagonist, administered orally for the prevention of postoperative vomiting after tonsillectomy in children. In a randomized, double-blind, placebo controlled study, 160 paediatric patients, ASA 1, aged 4-10 yr, received placebo or granisetron (20, 40 or 80 micrograms kg-1) (n = 40 each) orally, 1 h before surgery. A standard general anaesthetic technique was used throughout. A complete response, defined as no emesis and no need for another rescue antiemetic during the first 24 h after anaesthesia, occurred in 40%, 48%, 85% and 90% of patients who had received placebo, or granisetron 20, 40 or 80 micrograms kg-1, respectively (P < 0.05; overall Fisher's exact probability test). There were no clinically important adverse events. We conclude that preoperative oral granisetron, in doses more than 40 micrograms kg-1, was effective for the prevention of postoperative vomiting in children. PMID- 9861128 TI - Haemodynamic and electroencephalographic response to insertion of a cuffed oropharyngeal airway: comparison with the laryngeal mask airway. AB - We have compared the cuffed oropharyngeal airway (COPA), a modified Guedel airway device with a specially designed cuff at its distal end, with the laryngeal mask airway (LMA), on haemodynamic and electroencephalographic (EEG) responses to insertion. In addition, we examined the haemodynamic and EEG changes during initiation of the effect-compartment controlled infusion. We studied 35 female patients undergoing ambulatory gynaecological surgery allocated randomly to received an LMA or COPA to manage the airway. After premedication with midazolam 0.03 mg kg-1 i.v. and low-dose alfentanil (0.01 mg kg-1), anaesthesia was induced and maintained with propofol, using an effect-compartment controlled infusion set at an effect-site concentration of 4 micrograms ml-1. After intercompartmental equilibration, the LMA (group I) or COPA (group II) was inserted and haemodynamic (arterial pressure, heart rate) and EEG (bispectral index (BIS)) responses to insertion studied. The effect-compartment controlled infusion of propofol caused only mild haemodynamic changes during induction. Changes in arterial pressure and heart rate after insertion were similar in both groups and not significantly different from baseline values before insertion. Changes in BIS after insertion were minor and similar between groups. PMID- 9861129 TI - Saline as an alternative to air for filling the laryngeal mask airway cuff. AB - We have assessed a new method to evacuate saline completely from the laryngeal mask airway (LMA) cuff and tested the hypothesis that intracuff pressures, fibreoptic position and oropharyngeal leak pressures are similar for saline compared with air during nitrous oxide-oxygen anaesthesia. Eight size 4 LMA were inflated with saline 30 ml. After syringe evacuation, median residual weight was 0.56 (range 0.24-0.98) g; after additional manual cuff squeezing it was 0.26 (0.21-0.35) g; and after drying for 12 h at 60 degrees C with the valve open it was -0.02 (-0.05-0.04) g. Pressure-volume curves of four size 3-5 LMA showed that compliance was lower for the saline-filled cuff. A clinical study of 20 patients allocated randomly to have saline or air in the cuff showed a significant increase in intracuff pressure with air, but not saline, during nitrous oxide oxygen anaesthesia. The fibreoptic position of the LMA changed more frequently in the air, compared with the saline-filled group (four of 10 vs none of 10; P = 0.04). Oropharyngeal leak pressures were similar between groups. We conclude that the saline-inflated LMA cuff was reliably emptied and more stable in terms of intracuff pressures and possibly fibreoptic position. Filling the LMA cuff with saline is a viable option during laser surgery to the airway. PMID- 9861130 TI - Technique training: endoscopic percutaneous tracheostomy. AB - Percutaneous tracheostomy is being used increasingly in the intensive care unit and endoscopic control of this procedure affords an improved level of safety. Training in such new minimal access techniques can be a significant risk factor in patient outcome. Surgical simulation provides training which minimizes this risk. We present a method of training in percutaneous endoscopic tracheostomy using a simulation model based on animal tissue. Our experience with this model is reported. PMID- 9861131 TI - Evaluation of nitrogen dioxide scavengers during delivery of inhaled nitric oxide. AB - We have analysed the ability of three nitrogen dioxide absorbing materials (soda lime, noXon and zeolite) to act as nitrogen dioxide scavengers during delivery of inhaled nitric oxide. Different mixtures of gas were produced in a ventilator (Servo Ventilator 300) and passed through an inspiratory tube. Concentrations of nitrogen dioxide and nitric oxide were measured in the distal part of the tube, with and without the gas having passed through a canister containing the different filter materials. Our findings indicated that nitrogen dioxide was absorbed effectively by all filter materials but that there was re-formation of nitrogen dioxide from nitric oxide and oxygen in or immediately after the canister. This initial production of nitrogen dioxide was very rapid and could not be prevented by the use of scavengers. Thus soda lime and zeolite had no practical effect as scavengers in this delivery system, and the effect of noXon was very slight. PMID- 9861132 TI - In vitro degradation of atracurium and cisatracurium at pH 7.4 and 37 degrees C depends on the composition of the incubating solutions. AB - The pharmacokinetic models proposed for atracurium or cisatracurium are based on the assumption that spontaneous degradation via Hofmann elimination proceeds in vivo at the same rate as measured in vitro at pH 7.4 and 37 degrees C. As different degradation rates have been reported for all 10 stereoisomers of atracurium measured together, for each of its three isomeric groups, and for the single isomer cisatracurium, we studied if the rate is dependent on factors other than pH and temperature. In vitro degradation of atracurium and cisatracurium was studied at 37 degrees C and pH 7.4 in nine incubating solutions containing one of three buffer systems (phosphate, HEPES or Tris) and additives (sodium chloride, potassium sulphate or glucose). Concentrations of atracurium, cisatracurium and laudanosine were measured after incubation for up to 240 min using an HPLC method. Degradation of atracurium proceeded monoexponentially. The rate was slower in the presence of sodium chloride, potassium sulphate, and in a lower concentration of the phosphate buffer. Glucose enhanced the degradation. At the same total buffer concentration (50 mmol litre-1), degradation was fastest in the phosphate, intermediate in the HEPES and slowest in the Tris buffer. Degradation rates of cisatracurium in sodium phosphate 50 mmol litre-1 and Sorensen (Na-K phosphate) buffer 66.7 mmol litre-1 were similar to those of atracurium. We conclude that, at constant pH and temperature, the degradation rate of atracurium was dependent on the total concentration of the base in the incubating solution. PMID- 9861133 TI - I.v. anaesthesia and EEG burst suppression in rats: bolus injections and closed loop infusions. AB - We describe a system for monitoring and controlling i.v. anaesthesia in rats using burst suppression ratio (BSR) detection in the extradural EEG. After bolus injection, peak BSR values of 95% were achieved with propofol 8 mg kg-1, etomidate 3.5 mg kg-1 and alphaxalone 4.5 mg kg-1. Thiopental 32 mg kg-1 produced a peak BSR of 70% (larger doses were not tolerated). Recovery was fastest with propofol, followed by etomidate and alphaxalone with equal duration, and slowest with thiopental. In further experiments, a closed-loop infusion system maintained BSR accurately at targets of 30%, 50%, 70% or 90% for 60 min with propofol or etomidate. During these experiments the infusion rates were found to decrease with time, more so with etomidate (approximately 40%) than with propofol (approximately 20%). Recovery times were 2-3 times longer with etomidate than with propofol. This model demonstrated differences between i.v. anaesthetics and may be useful in screening new compounds in preclinical development. PMID- 9861134 TI - Steady-state propofol brain:plasma and brain:blood partition coefficients and the effect-site equilibration paradox. AB - Based on volume-flow relationships, CNS agents that are highly lipid soluble (log octanol-water partition coefficient > 2) are expected to have equilibration half times (T1/2 kE0) that are proportional to brain solubility. Propofol, the most lipophilic anaesthetic in clinical use, has T1/2 kE0 values of 1.7 and 2.9 min in rats and humans, respectively, compared with an expected value of at least 8 min. As a first step in exploring this discrepancy between observed and predicted values, we determined the steady state brain:plasma and brain:blood partition coefficients in rats after a 4-h infusion of propofol. Brain:plasma and brain:blood partition coefficients were 8.2 (SD 1.6) and 3.0 (0.5), respectively. T1/2 kE0 predictions based on brain: blood partitioning in rats are more in agreement with the observed equilibration half-time, suggesting that drug bound to the formed elements of blood participates in the uptake and transfer of propofol to its effect site. PMID- 9861136 TI - Effects of etomidate, propofol and thiopental anaesthesia on arteriolar tone in the rat diaphragm. AB - We have assessed, by intravital microscopy in rats, the effects of different anaesthetics on diaphragmatic arteriolar diameter. Rats were anaesthetized with etomidate, propofol or thiopental (groups E, P and T, respectively) and the diameters of the arterioles were measured sequentially at baseline and after topical application of either mefenamic acid (MA, 20 mumol litre-1) or N omega nitro-L-arginine (NNA, 300 mumol litre-1), inhibitors of prostaglandins and nitric oxide, respectively. In group E, baseline arteriolar diameters were significantly higher than those in the two other groups (P < 0.01). MA and NNA induced significant constriction in the three groups (P < 0.001). However, whereas constriction induced by NNA was similar in the three groups, constriction induced by MA was significantly higher in group E compared with groups P and T (P < 0.05). We conclude that diaphragmatic arteriolar diameters in rats were greater during etomidate than during thiopental or propofol anaesthesia. This phenomenon may be mediated by prostaglandins. PMID- 9861135 TI - Effects of posture on blood flow diversion by hypoxic pulmonary vasoconstriction in dogs. AB - We used differential excretion of sulphur hexafluoride from the left and right lung to measure blood flow diversion by hypoxic pulmonary vasoconstriction (HPV) in the prone and supine positions in dogs (n = 9). Gas exchange was assessed using the multiple inert gas elimination technique. Blood flow diversion from the hypoxic (3% oxygen) left lung was mean 70.7 (SD 11.2)% in the supine compared with 57.0 (12.1)% in the prone position (P < 0.02). The supine position was associated with increased perfusion to low VA/Q regions (P < 0.05). The increased flow diversion with hypoxia in the supine position was associated with more ventilation to high VA/Q regions (P < 0.05). We conclude that flow diversion by hypoxic pulmonary vasoconstriction is greater in the supine position. This effect could contribute to the variable response in gas exchange with positioning in patients with ARDS. PMID- 9861137 TI - Primary afferent-evoked release of immunoreactive galanin in the spinal cord of the neuropathic rat. AB - We have determined if peripheral nerve stimulation altered the increased spontaneous release of immunoreactive (ir)-galanin that is found in the superficial dorsal horn of the spinal cord of neuropathic rats. Using the antibody microprobe technique to study the localized sites of ir-galanin release in vivo, we found that high intensity electrical stimulation of the injured nerve resulted in a further increase in ir-galanin release in the superficial dorsal horn, with no significant persistence of ir-galanin after release. Release of ir galanin at stimulus strengths sufficient to activate C fibres, in an area of the spinal cord thought to be concerned with nociceptive transmission, indicates a possible role for this peptide in the spinal modulation of pain after peripheral nerve injury. PMID- 9861138 TI - Antibacterial activity of lidocaine in mixtures with Diprivan. AB - We have studied the antibacterial activity of different concentrations of 0.005 2% lidocaine (lignocaine) in mixtures with Diprivan (propofol), against micro organisms commonly implicated in sepsis as a result of extrinsically contaminated Diprivan. Bacterial colony counts were reduced progressively with increasing concentrations of lidocaine. Bacteriostatic and bactericidal concentrations of lidocaine were 0.2-2%. Lidocaine 2% was not bactericidal for one of the seven organisms tested. By inhibiting bacterial replication, lidocaine, when added to Diprivan to reduce pain on injection, may possibly reduce the harmful consequences if extrinsic contamination occurs. PMID- 9861139 TI - Postoperative cognitive deficit in the elderly surgical patient. PMID- 9861140 TI - Mivacurium compared with succinylcholine in children with liver disease. AB - We have compared mivacurium and succinylcholine in 27 paediatric patients with mild (Child's A) to moderate (Child's B) liver disease undergoing oesophagogastroduodenoscopy (OGD) and injection of oesophageal varices, with 10 healthy children receiving mivacurium for ENT procedures. With mivacurium 0.2 mg kg-1, the severity of liver disease did not correlate with duration of block compared with controls (time from bolus to T1 25%, P = 0.74; T1 25% to T4:T1 > 0.7, P = 0.545). However, initial recovery (time to T1 25%, P = 0.002) and overall recovery (bolus to T4:T1 > 0.7, P = 0.004) from mivacurium-induced neuromuscular block correlated inversely with pre-existing concentrations of plasma cholinesterase. Conditions for tracheal intubation at 2 min with mivacurium were comparable with conditions at 1 min with succinylcholine in the liver patients. PMID- 9861141 TI - Atropine premedication and the cardiovascular response to electroconvulsive therapy. AB - A report by the Royal College of Psychiatrists recommended avoiding atropine premedication during electroconvulsive therapy (ECT). We have examined the cardiovascular effects of ECT with or without atropine premedication. Consenting patients (n = 30) were allocated randomly before their third ECT session to receive atropine or no premedication. The rate pressure product (RPP) was recorded before anaesthesia, before ECT stimulus and at 1-min intervals thereafter for 5 min. Patients who did not receive atropine had significantly lower RPP values after all stimulus recordings. Administration of atropine or not explained 32% of the variance of summated RPP after the stimulus. There was no clinically significant bradyarrhythmia in those who did not receive atropine. Our findings support the recommendation of the Royal College of Psychiatrists. The study suggests that when threshold determination is not needed, avoiding atropine effectively contains potentially harmful cardiovascular responses. PMID- 9861142 TI - Adrenal failure in the critically ill. AB - Three critically ill patients suffered multiple organ failure secondary to sepsis. Despite adequate supportive therapy and appropriate antibiotic cover, they failed to improve and required inappropriate inotrope support. They had not been treated with steroids or other drugs known to suppress adrenal function. Adrenal insufficiency was suspected. A random cortisol concentration and a short synacthen test demonstrated concentrations below the range expected in all three cases. High-dose steroid therapy was commenced with marked improvement in the short-term. However, in each case sepsis eventually caused death. PMID- 9861143 TI - Repeated transient neurological symptoms after spinal anaesthesia with hyperbaric 5% lidocaine. AB - We report a case of repeated delayed pain after cystoscopy under spinal lidocaine anaesthesia, which may be caused by transient radicular irritation. The possible aetiology of the symptoms is discussed. PMID- 9861144 TI - Prolonged endotracheal intubation in infants and children. 1965. PMID- 9861145 TI - Anaesthesia for telescopic procedures in the thorax. PMID- 9861146 TI - Anaesthesia for telescopic procedures in the thorax. PMID- 9861147 TI - A case made for automated anaesthetic record keeping. PMID- 9861148 TI - Peripheral blocks of the lower limb for repair of fractured neck of femur. PMID- 9861149 TI - Propofol and asepsis: is it safer to use the TCI technique? PMID- 9861150 TI - Ketorolac does not increase the risk of renal dysfunction after lung surgery. PMID- 9861151 TI - Local anaesthesia for routine ocular surgery. PMID- 9861152 TI - Pain after amputation. PMID- 9861153 TI - Stature of anaesthetic personnel and positioning of patients. PMID- 9861154 TI - Vecuronium, like rocuronium, causes pain on injection. PMID- 9861155 TI - Involvement of meiotic resumption in the disruption of gap junctions between cumulus cells attached to pig oocytes. AB - The present study was undertaken to examine the mechanisms by which cumulus cells regulate meiotic resumption in pig oocytes using microinjections of lucifer yellow into cumulus-oocyte complexes combined with a fluorescent assay. Some cumulus-oocyte complexes cultured for 0, 8, 16, 24, 32, 40 and 48 h were denuded to assess the nuclear status of oocytes; the remaining complexes were injected with lucifer yellow and monitored for the transfer from the oocyte to the surrounding cumulus cells using confocal laser scanning microscopy. The proportion of oocytes undergoing germinal vesicle breakdown at 16, 24 and 32 h of cultivation was much higher than that of cumulus-oocyte complexes in which all gap junctions within cumulus cells and between cumulus cells and oocyte were disrupted (16 h: 22.7% versus 8.8%, 24 h: 66.7% versus 40.3%, 32 h: 84.0% versus 69.4%), showing that the disruption does not trigger meiotic resumption in the pig oocyte. A significant positive correlation (r = 0.99, P < 0.01) was established between the proportion of germinal vesicle breakdown oocytes and that of cumulus-oocyte complexes exhibiting loss of gap junctions within all cumulus cell layers excluding the innermost layer. From these results, it is concluded that meiotic resumption in pig oocytes is induced by the disruption of gap junctions within cumulus cells, rather than that between the oocyte and cumulus cells, which blocks the conduction of meiosis inhibitory signals from the outer cumulus cells to the oocyte. PMID- 9861156 TI - Endometrial oxytocin receptor and uterine prostaglandin secretion in mares during the oestrous cycle and early pregnancy. AB - Circulating concentrations of 13,14-dihydro-15-ketoprostaglandin F2 alpha (PGFM) were measured before and after administration of oxytocin and after endometrial biopsy, with or without uterine flushing performed per vaginam, on days 10, 14 and 18 after ovulation in nine pregnant and nine cyclic mares. Concentrations of oxytocin receptor were measured in endometrial biopsy samples. Neither pregnancy status nor time after ovulation affected basal PGFM concentrations. PGFM concentrations were increased after oxytocin administration on each of the days studied in cyclic mares; on day 14 the mean response was 4.5 times higher than the mean response on days 10 and 18. In contrast, during pregnancy, responses to oxytocin administration occurred only on days 10 and 18. Marked increases in PGFM concentrations in response to endometrial biopsy occurred only on day 14 in cyclic mares and on day 18 in pregnant mares. Mean concentrations of oxytocin receptor were between 200 and 300 fmol mg-1 protein on day 10 in both pregnant and cyclic mares; in cyclic mares oxytocin receptor concentrations were increased approximately threefold on day 14 compared with days 10 and 18, but no such increase was evident during pregnancy. Total amounts of PGFM secreted after oxytocin treatment correlated with endometrial oxytocin receptor concentrations in cyclic (P < 0.001) but not in pregnant (P > 0.5) mares, and the same was true for PGFM release induced by endometrial biopsy (cyclic: P = 0.0025; pregnant: P > 0.5). The data support the hypothesis that endometrial concentrations of oxytocin receptor determine uterine prostaglandin F2 alpha secretion in cyclic mares and that endometrial oxytocin receptor concentrations are reduced in early pregnancy by a product of the conceptus. The increase in response of the pregnant uterus to oxytocin treatment or biopsy-flushing between days 14 and 18 was not due to an increase in the concentration of oxytocin receptors but presumably reflected increased receptor sensitivity. PMID- 9861157 TI - Not only inner cell mass cell nuclei but also trophectoderm nuclei of mouse blastocysts have a developmental totipotency. AB - The nuclei of mouse trophectoderm cells were found to have developmental totipotency like inner cell mass cells after serial nuclear transfer. Single inner cell mass or trophectoderm cells from expanded blastocysts synchronized with the cell cycle by treatment with nocodazole and aphidicolin to the G1 stage were injected into the perivitelline space of enucleated metaphase II oocytes together with Sendai virus. All oocytes were given three electrical pulses to induce fusion and activation (first nuclear transfer). Aphidicolin was present in all media used until fusion. When reconstituted oocytes developed to the two-cell stage, the nuclei of the reconstituted eggs were fused with the enucleated blastomeres of fertilized two-cell embryos by inactivated Sendai virus (second nuclear transfer). The reconstituted embryos were cultured in vitro and transferred to recipients. After the second nuclear transfer, 23-64% (for inner cell mass cells) and 32-62% (for trophectoderm cells) developed to morula or blastocyst stage. Better development of second nuclear transfer embryos was observed when oocytes fused with trophectoderm nuclei did not extrude a polar body after the first nuclear transfer. After transfer of morulae and blastocysts to recipients, four males were obtained, two from inner cell mass and two from trophectoderm nuclei. These findings indicate that the nucleus of inner cell mass and trophectoderm cells of mouse blastocysts can be reprogrammed within the cytoplasm of unfertilized oocytes and then in fertilized embryos. PMID- 9861158 TI - Differential effects of ovarian local anaesthesia during pro-oestrus on ovulation by the right or left ovary in normal and hemi-ovariectomized adult rats. AB - The effects on spontaneous ovulation in normal and hemi-ovariectomized cyclic rats of an injection of saline or a local anaesthetic (2-diethylamine-2',6' acetoxilidide hydrochloride with adrenaline, Xylocaine; administered at 13:00 h) into the bursa ovarica, were studied to investigate the participation of ovarian innervation in the regulation of the response of ovarian follicles to endogenous gonadotrophins on the day of pro-oestrus. The injection of saline or Xylocaine into both ovaries did not affect ovulation by the right ovary. Bilateral injection of Xylocaine did not affect ovulation by the left ovary. However, when saline was injected into the right ovary and Xylocaine into the left ovary, the number of ova shed by the left ovary decreased significantly (0.9 +/- 0.7 versus 4.4 +/- 0.2, P < 0.01; Mann-Whitney U test). In hemi-ovariectomized animals, when the right ovary was anaesthetized, ovulation was similar to that in saline treated animals, and when the right ovary was anaesthetized, ovulation was blocked (one of six animals ovulated). The injection of hCG at 14:00 h to hemi ovariectomized rats did not restore ovulation, whereas all the rats ovulated when anaesthesia occurred at 18:00 h. The present results suggest that on the afternoon of the day of pro-oestrus, before the LH peak, a neural signal modulates the response of the left ovary to gonadotrophins. PMID- 9861159 TI - Improved method of superovulation in monovulatory brushtail possums (Trichosurus vulpecula) using pregnant mares' serum gonadotrophin-luteinizing hormone. AB - A new superovulation regimen for the monovulatory brushtail possum (Trichosurus vulpecula) has been devised. It reduces the number of hormone treatments required and elicits a better rate of ovulation than the established pregnant mares' serum gonadotrophin (PMSG)-GnRH method. Ovarian stimulation was achieved by a single intramuscular injection of 15 iu PMSG. This treatment resulted in the recruitment and development of a large number (9-12) of Graafian follicles on the ovaries. Induction of ovulation was achieved by a single intramuscular injection of 2-10 mg pig LH or multiple intramuscular injections of GnRH, (4 x 50 micrograms, 90 min apart) given 72 h after PMSG treatment. Superovulation occurred in all animals (n = 48) examined 48 h after treatment irrespective of dose of LH or type of ovulatory stimulus used. The highest ovulatory success (83%), the maximum number of ovulation sites (9.5 +/- 2.8) and the highest number of oocytes recovered (9.0 +/- 2.5) were achieved after treatment with PMSG and 4 mg LH. These results were significantly greater than the ovulatory success (43%), numbers of ovulation sites (3.9 +/- 1.1) and number of oocytes recovered (2.1 +/- 0.9) after PMSG-GnRH treatment (P < 0.05). This simpler and more effective superovulation protocol should assist with more effective manipulation of possum reproduction in captivity. PMID- 9861160 TI - Long-term, quantitative analysis of gametogenesis in autotriploid rainbow trout, Oncorhynchus mykiss. AB - A long-term, quantitative analysis was conducted on the gametogenesis of autotriploid rainbow trout (Oncorhynchus mykiss) to quantify their degree of germline development and reproductive potential. Triploid and diploid (control) trout siblings were raised separately under identical conditions and sampled randomly for histological analysis. Triploid males underwent testicular development and proliferation of germ cells by mitosis and meiosis, progressing through initial phases of spermatogenesis at a similar pace to diploid controls. The effects of triploidy on males were most evident during the final stages of spermatogenesis, when all diploid males contained free spermatozoa in the lumen of most tubules (average relative frequency, ARF = 68.5%), whereas triploid males contained predominantly spermatocytes (ARF = 36.3%) and morphologically abnormal spermatozoa (ARF = 31.8%). In contrast, the gonadal development of triploid females was affected during its early stages; the major patterns observed were the arrest of the oogonia within oogonial clusters (ARF = 30.4-71.1%), the appearance of small numbers (ARF = 1.5-6.0%) of previtellogenic and early vitellogenic follicles, and the proliferation of non-follicular elements (vascular lacunae, fibrosis and tubular adenomas). In agreement with previous reports on the ovarian development of chromosomally female (3A:ZZW) triploid chickens, male-differentiating areas (ARF = 0.2-12.2%) were observed in most triploid females examined, which by the end of the sampling period appeared as gonadal hermaphrodites. It is hypothesized that the lack of proper somatic-to germ cell interactions prevents the segregation of the oocytes from the gonial clusters and may explain the early blockage observed during the gonadal morphogenesis of autotriploid female rainbow trout. PMID- 9861161 TI - Analysis of the mitotic index of chicken primordial germ cells before and after settling in the germinal ridge. AB - The mitotic index of chicken primordial germ cells was determined by analysing the nuclear DNA content and the number of M phase and S phase primordial germ cells at stage 14-15 (before settling in the germinal ridge) and at stage 17-18 (after settling in the germinal ridge) using an interactive laser cytometer. When nuclear DNA content was presented graphically, two peaks were revealed at each of the two stages being studied. The value of the second peak was approximately twice that of the first peak. Primordial germ cells, having two nuclei in one cell, were recognized at each stage and were considered to be at M phase in cell division. The amount of nuclear DNA in both S and M phases was correlated with that between the first and the second peaks and that in the second peak, respectively. The number of M phase and S phase primordial germ cells significantly increased after they had settled in the germinal ridge. These results suggest that chicken primordial germ cells start to proliferate actively as soon as they reach the gonadal ridge. PMID- 9861162 TI - Morphometric and endocrine analyses of the effects of nutrition on the testis of mature Merino rams. AB - The effects of nutrition on the testis were investigated in groups of five mature Merino rams that were fed either a sub-maintenance (low) diet or a supra maintenance (high) diet for 69 days. Testosterone, oestradiol and inhibin were measured in blood plasma sampled simultaneously from jugular and testicular veins after an i.v. injection of 200 ng ovine LH kg-1. Plasma concentrations of testosterone, inhibin and oestradiol were higher in testicular than in jugular vein plasma for both diets (P < 0.01). After the LH injection, jugular plasma testosterone increased more rapidly (P < 0.01) in rams fed the high diet than in rams fed the low diet. This was not seen in the testicular vein. Oestradiol concentrations were higher in rams on the high diet than in those on the low diet, in both the testicular (P < 0.0001) and the jugular vein (P < 0.02). Diet did not affect inhibin concentrations. Testes were surgically removed and processed for light microscopy. Testicular mass and seminiferous tubule length and diameter were higher with the high diet than the low diet (P < 0.01). The number of Sertoli cell nuclei per testis was also affected (high diet: 120 +/- 6 x 10(8); low diet: 77 +/- 7 x 10(8); P < 0.001), whereas the proportion of testis occupied by Sertoli cell nuclei was not affected. The number of Leydig cells per testis was not affected by diet, but Leydig cells occupied a greater volume of testis in rams on the high diet than in those on the low diet (P < 0.001). The effects of nutrition on Leydig and Sertoli cells are consistent with changes in the endocrine and exocrine functions of the testis. The finding that Sertoli cell population was altered in adult rams may be explained by the GnRH-independent effects of nutrition. PMID- 9861163 TI - Microscopic analysis of enzyme activity, mitochondrial distribution and hydrogen peroxide in two-cell rat embryos. AB - A developmental block is induced by phosphate in rat embryos at the late two-cell stage. The present study was designed to examine the energy metabolism of rat two cell blocked and non-blocked embryos. Enzyme activity was measured in individual embryos by histochemical techniques. The activities of malate dehydrogenase, isocitrate dehydrogenase, lactate dehydrogenase, pyruvate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, glutamate dehydrogenase, glucose-6 phosphate dehydrogenase, glucose-6-phosphatase, and phosphorylase did not differ among non-blocked and blocked embryos. However, the activity of succinate dehydrogenase was significantly decreased in blocked embryos compared with non blocked embryos. In blocked embryos, cytochrome oxidase activity was distributed homogeneously, but was located at the perinuclear region in non-blocked embryos. Active mitochondrial organization was visualized using the fluorescent probe rhodamine 123 and laser scanning confocal microscopy. In both non-blocked and blocked embryos, mitochondria were distributed homogeneously. The concentration of H2O2 measured fluorometrically in embryos cultured without phosphate did not change significantly during the culture period, but decreased in embryos cultured with phosphate. The timing corresponded to the occurrence of the two-cell block. In summary, these results suggest that the developmental block in rat two-cell embryos is induced by disturbance of mitochondrial energy metabolism. PMID- 9861164 TI - Effects of thyroidectomy and thyroxine replacement on seasonal reproduction in the red deer hind. AB - Two experiments were conducted to test whether thyroid hormones are required for the cessation of the breeding season in female red deer. In Expt 1, 16 mature hinds were allocated early in the breeding season (May) to the following groups: thyroidectomized (THX), thyroidectomized and treated with subcutaneous thyroxine implants which increased plasma tri-iodothyronine to physiological concentrations (THX + T4), or euthyroid controls. Plasma progesterone concentrations consistent with oestrous cyclicity were evident in all hinds during the breeding season (May September) but after 23 September plasma progesterone concentrations became generally low (< 2.0 nmol l-1) in control and THX + T4 hinds. In contrast, thyroidectomized hinds not receiving thyroxine continued to exhibit circulating progesterone comparable with breeding season concentrations until the end of the experiment (late December). Nine hinds were ovariectomized or ovariectomized and thyroidectomized in May and treated with subcutaneous oestradiol-impregnated capsules, which were removed for periods of about 1 months during August, November and January, to test whether thyroid hormones are involved in the seasonal decline of LH secretion (Expt 2). In the presence of oestradiol, basal LH concentrations and episodic LH secretion were low during the non-breeding season (September-March) (P < 0.05). During this season, basal and GnRH-induced LH concentrations and LH pulse frequency remained similar for both groups except in the absence of oestradiol, when basal LH (3.1 +/- 0.8 versus 1.3 +/- 0.5 ng ml 1, P < 0.05 and GnRH-induced LH (25.9 +/- 3.6 versus 4.7 +/- 0.4 ng ml-1, P < 0.001) concentrations were higher in thyroidectomized hinds than in euthyroid hinds, respectively. Twenty-five hinds were ovariectomized and thyroidectomized during the breeding season and treated with subcutaneous thyroxine implants at different times to identify when thyroid hormones are required to be present to suppress LH concentrations during the non-breeding season in the absence of oestradiol. Thyroxine treatment at the beginning of or during the non-breeding season was effective in suppressing plasma LH concentration, but this action of thyroid hormones did not occur during the breeding season. These results indicate that, in red deer hinds thyroxine is required for termination of the breeding season and that thyroid gland secretions specifically block steroid-independent inhibition of reproductive activity during the non-breeding season. This inhibitory mechanism requires thyroid hormones to be present only from around the time of the end of the breeding season for their normal expression, and they remain responsive to thyroid hormones after this period. PMID- 9861166 TI - Isolation of proteins from subacrosomal region of spermatozoa from a marsupial, the tammar wallaby (Macropus eugenii). AB - Recent studies indicate that subacrosomal proteins are necessary for the attachment of the acrosome onto the nucleus during sperm formation, and for the stability of the nuclear membrane during fertilization. For the first time, subacrosomal proteins have been isolated from a marsupial species, the tammar wallaby (Macropus eugenii), using a method developed in our laboratory. Whole ejaculated spermatozoa were fractionated into head and tail sections by ultrasonication to extract subacrosomal proteins. The heads (> 95% purity) were then isolated from tail sections using centrifugation with a three-step discontinuous sucrose gradient (35, 68 and 75% (w/v). The heads were treated with 0.1% (v/v) Triton X-100 which stripped off the acrosome, but not the subacrosomal proteins, from the head. The proteins were finally extracted by 100 mmol NaOH l 1. Four prominent subacrosomal polypeptides, with molecular masses of 45, 38, 33 and 29 kDa, were recognized from the SDS-PAGE gel. The localization of these polypeptides (particularly the 45 kDa polypeptide) was confirmed by fluorescent and immunogold labelling with polyclonal antibodies raised in mice against the obtained polypeptides. In wallaby testes, the 45 kDa polypeptide was detected as early as at the step 3 spermatid and was mainly associated with the membrane of the newly formed acrosome vesicle. This polypeptide was also found on the acrosomal membrane of all older spermatids. The 45 kDa polypeptide was found on the acrosomal region of the spermatozoa collected from the caput, corpus and cauda of the epididymis. The similarity of the sperm anatomy of the tammar wallaby with that of other marsupials, such as the brushtail possum, implies that this procedure could be applied effectively to other marsupial species with minor modification. PMID- 9861165 TI - Regulation of ovine GnRH receptor gene expression by progesterone and oestradiol. AB - The objective of this study was to determine whether progesterone prevents the stimulatory effects of oestradiol on GnRH receptor gene expression. In Expt 1, ewes were treated during the luteal phase (days 10-12 of the oestrous cycle) with either one or five subcutaneous implants containing oestradiol (n = 6 per group). Control ewes received no treatment (n = 6). Anterior pituitary glands were collected 16 h after treatment with oestradiol. Steady-state amounts of GnRH receptor mRNA were similar among all three treatment groups despite increased circulating concentrations of oestradiol in implanted ewes at the time of pituitary collection (4.3 +/- 0.6 and 24.7 +/- 2.6 pg ml-1 in ewes treated with one or five implants, respectively, compared with 0.5 pg ml-1 in controls). Experiment 2 was designed to determine whether progesterone was the ovarian factor preventing the stimulatory effects of oestradiol on expression of the GnRH receptor gene in Expt 1. Twenty-five ewes were ovariectomized on day 6 or day 7 of the oestrous cycle and assigned to one of five treatment groups (n = 5 per group). Control ewes received no further treatment. Endogenous luteal phase concentrations of progesterone were replaced in three groups of ewes at the time of ovariectomy via intravaginal implants. Three days after ovariectomy, one group of progesterone-treated ewes received one oestradiol implant, while another group of progesterone-treated ewes received five oestradiol implants. An additional group was treated with five oestradiol implants only, and anterior pituitary glands were collected from all ewes 16 h later. Compared with untreated ovariectomized ewes, treatment with progesterone alone did not affect amounts of GnRH receptor mRNA. In ewes treated with progesterone and either one or five oestradiol implants, steady-state amounts of GnRH receptor mRNA were increased twofold (P < 0.01). Treatment with oestradiol in the absence of progesterone increased amounts of GnRH receptor mRNA threefold (P < 0.001). These results provide evidence that the stimulatory effects of oestradiol on the expression of the GnRH receptor gene are prevented during the natural luteal phase in ewes. However, progesterone does not appear to act independently to mediate this effect. PMID- 9861167 TI - Sexual differences in the role of kainate receptors in controlling gonadotrophin secretion in prepubertal rats. AB - Glutamate stimulates LH secretion in adult female rats after activation of N methyl-D-aspartic acid (NMDA), kainate, and 2-amino-3-hydroxy-5-methyl-4-isoxazol propionic acid receptors. In contrast to the positive role of kainate receptors in the control of LH secretion in adult females, neither activation nor antagonization of kainate receptors in immature rats modified the onset of puberty. The present experiments were carried out to establish why, if kainate stimulates LH release in adult rats, it fails to advance puberty in immature rats, and to determine whether the role of kainate receptors is sexually dimorphic around puberty. In Expt 1, 4-, 8-, 12-, 16-, 20- and 30-day-old females were investigated 15 min after administration of vehicle or kainic acid, a kainate receptor agonist (2.5 mg kg-1). In Expt 2, 30-day-old female rats were studied 2, 5 and 10 min after administration of vehicle, 2.5 mg kainic acid kg-1 or NMDA, an NMDA receptor agonist (15 mg kg-1). In Expt 3, female and male rats were gonadectomized or sham-gonadectomized on day 23 and investigated on day 30 after injection of vehicle, kainic acid (2.5 mg kg-1 at -15 min) or 6,7 dinitroquinoxaline-2,3 dione (DNQX), a kainate antagonist (2 mg kg-1 in two injections at -120 and -60 min). Finally, 30-day-old female and male rats were investigated 15 min after injection of vehicle or NMDA (15 mg kg-1) or 60 min after administration of different doses of 5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine (MK-801), an NMDA antagonist (0.1, 0.25 or 0.50 mg kg-1). The results indicate that the role of kainate receptors in the control of gonadotrophin secretion is sexually dimorphic around puberty, since: (a) LH secretion was stimulated by kainic acid in male rats but inhibited in females; (b) FSH secretion was inhibited by kainic acid in ovariectomized females, but not in orchidectomized males; and (c) DNQX inhibited LH secretion in males but not in females. These differences were specific for kainate receptors since, in both sexes, NMDA stimulated and MK-801 inhibited LH secretion. It may be concluded that the secretion of gonadotrophins is modulated differently by kainate receptors in prepubertal male and female rats. PMID- 9861168 TI - Ovarian follicular dynamics during anoestrus in ewes. AB - The aim of the present study was to document ovarian antral follicle dynamics throughout seasonal anoestrus in sheep. Daily transrectal ultrasonography was performed during four 17 day scanning periods from March to July in Western White faced crossbred ewes. Blood samples were collected each day with ultrasonographic scanning for measurement of serum concentrations of FSH, oestradiol and progesterone. Blood samples were also taken every 15 min for 6 h, mid-way through each period of ultrasonographic examination, to determine the patterns of secretion of gonadotrophic hormones. Hormonal data were then related to observed changes in follicular populations and the patterns of antral ovarian follicle turnover. Ultrasonography showed that the ovaries of anoestrous ewes remained active and that the largest ovarian antral follicles grew to a periovulatory size (> or = 5 mm in diameter) at all stages of anoestrus. The total number of all ovarian follicles > or = 3 mm in diameter was lower during early anoestrus compared with at mid-anoestrus because of a significantly smaller number of small (3 mm) and medium (4 mm) ovarian follicles. The largest ovarian follicles (attaining > or = 5 mm in diameter before regression) exhibited a wave-like pattern of growth; an average of three waves of follicular development were recorded in sheep during each of the four 17 day scanning periods in anoestrus, with follicular waves emerging approximately every 5 days. This rhythmic pattern of follicular emergence was found to be associated with the occurrence of fluctuations in serum FSH concentrations. The growth rate of the largest follicles of the wave increased significantly from early to late anoestrus in sheep. In addition, ovarian follicles not growing beyond 3 mm in diameter showed organized patterns of growth and regression; their numbers tended to be lower (P = 0.09) at 3 days before and on the day of follicular wave emergence. Some ewes were seen to maintain synthesis of progesterone throughout anoestrus. This submaximal progesterone secretion tended to occur at irregular intervals and was not coupled with changes in concentrations or patterns of gonadotrophin release, ovulations or detectable morphological luteinization of ovarian antral follicles. It was concluded that the growth of ovarian antral follicles to an ovulatory size was maintained throughout anoestrus in ewes, with a transient shift in the number of small and medium-sized follicles during mid-anoestrus, and that the periodic emergence of waves of large follicles (> or = 5 mm in diameter) occurred in synchrony with an endogenous rhythm of FSH secretion. PMID- 9861169 TI - Patterns of growth, oestradiol and progesterone released by in vitro cultured mouse ovarian follicles indicate consecutive selective events during follicle development. AB - This study investigated the relationship between individual follicle growth, steroid release and follicular morphology to provide basic information about critical stages in follicle development. Preantral mouse follicles secreted significant and constant amounts of progesterone that were not related to oestradiol production but did appear to be related to thecal organization. Oestradiol release was variable among follicles of equivalent diameter, but marked increases in oestradiol concentrations were measured in follicles of 300 and 400 microns diameter. Over 4 days of culture, the proportion of follicles growing beyond the threshold diameter of 300 microns was lower in small (140-160 microns) than it was in large preantral follicles (170-210 microns). Retardation of growth below this diameter was associated with significantly decreased steroid concentrations. In follicles growing beyond the threshold diameter, antrum formation progressed, but full Graafian stage was not attained. Among large follicles, variability in oestradiol release could not be associated with obvious histological differences. In contrast, structural disturbances were associated with reduced steroid production and growth abnormalities. Therefore, it is concluded that the preantral follicles selected for culture comprise distinct follicle populations with respect to their developmental potential in vitro, and that follicle development is controlled by endogenous processes involving oestradiol at distinct stages of follicle development. PMID- 9861170 TI - Steady-state concentrations of mRNA encoding two inhibitors of protein kinase C in ovine luteal tissue. AB - Prostaglandin F2 alpha (PGF2 alpha) decreases secretion of progesterone from the corpus luteum in domestic ruminants. However, it is less effective during the early part of the oestrous cycle (Louis et al., 1973) and at the time of maternal recognition of pregnancy (Silvia and Niswender, 1984; Lacroix and Kann, 1986). Decreased luteal responsiveness may be due to failure of PGF2 alpha to activate fully its normal second messenger system, protein kinase C (PKC). Alternatively, increased resistance of the corpus luteum to PGF2 alpha might be attributable to greater concentrations of recently identified biological inhibitors of PKC. These possibilities were addressed by measuring steady-state concentrations of mRNA encoding PGF2 alpha receptor and two inhibitors of PKC, protein kinase C inhibitor-1 (PKCI-1) and kinase C inhibitor protein-1 (KCIP-1, brain 14-3-3 protein), in corpora lutea collected from ewes on days 4, 10 and 15 of the oestrous cycle (n = 5 per day) and day 15 of pregnancy (n = 7). There were no differences in mean concentrations of mRNA encoding PGF2 alpha receptor among the groups. However, concentrations of mRNA encoding both inhibitors of PKC were higher (P < 0.01) on day 4 of the oestrous cycle compared with the other groups. Treatment of ewes with a luteolytic dose of PGF2 alpha, which activates PKC, did not change concentrations of mRNA encoding either PKCI-1 or KCIP-I up to 24 h later. Luteal expression of mRNA encoding the PKC inhibitors and PGF2 alpha receptor was also examined in ewes treated with oestradiol in vivo for 16 h in the midluteal phase. High concentrations of oestradiol in serum (20 and 70 pg ml 1) did not influence quantities of any of the mRNAs examined. Therefore, an increase in PKC inhibitors may be involved in resistance of the corpus luteum to PGF2 alpha during the early part of the oestrous cycle but does not appear to mediate the increased resistance of the corpus luteum to PGF2 alpha during maternal recognition of pregnancy. Neither PGF2 alpha nor oestradiol affected steady-state concentrations of mRNAs encoding PKCI-1 or KCIP-I. PMID- 9861171 TI - Spontaneous testicular regression in collared lemmings (Dicrostonyx groenlandicus) exposed to prolonged long photoperiod. AB - Reproductive maturation and attainment of maximal gonadal size in collared lemmings (Dicrostonyx groenlandicus) is facilitated by transfer from a short photoperiod (8 h light: 16 h dark) to a long photoperiod (22 h light:2 h dark). However, reproductive maturation in lemmings born in 22 h light:2 h dark is impeded by exposure to the natal photoperiod of 22 h light:2 h dark. Data from adult lemmings suggest that prolonged exposure to a long photoperiod of 20 h light:4 h dark results in gonadal regression. The present experiments were conducted to test the hypothesis that adult lemmings exposed to a prolonged long photoperiod undergo testicular regression. Male collared lemmings were transferred at weaning (19 days of age) from the natal photoperiod of 8 h light:16 h dark to one of two long photoperiods: 22 h light:2 h dark or 24 h light:0 h dark (constant light). Gonadal mass was maximal in both groups 10 weeks after weaning. However, 20 weeks after weaning, gonadal mass in both groups was low (relative to that at 10 weeks), suggesting that regression had occurred. When testes mass was maximal 10 weeks after weaning, animals were transferred in groups from 22 h light:2 h dark to one of several shorter photoperiods: 20 h light:4 h dark, 18 h light:6 h dark, 16 h light:8 h dark, or 8 h light:16 h dark. Testes mass 20 weeks after weaning was greatest, that is, not different from maximal, in animals from the 20 h light:4 h dark and 18 h light:6 h dark groups, intermediate in animals from the 16 h light:8 h dark group, and low in animals from the 8 h light:16 h dark group. These observations indicate that transfer from 22 h light:2 h dark to a slightly shorter photoperiod permits maintenance of gonadal size, but transfer to a short photoperiod causes testicular regression. Body weight, bifid claw width and pelage colour did not change during spontaneous gonadal regression in animals housed in a long photoperiod for 20 weeks. These results suggest that prolonged exposure to an unchanging long photoperiod causes spontaneous testicular regression, and that photoperiod regulates reproductive and somatic parameters independently. PMID- 9861172 TI - Gonadotrophin profiles and dioestrous pulsatile release patterns in mares as determined by collection of jugular blood at 4 h intervals throughout an oestrous cycle. AB - In mares, dioestrous FSH profiles based on once-a-day sampling are variable; however, the pulsatility of plasma FSH, which has been suggested by limited windows of intensive sampling, may contribute to this variability. Jugular blood from six mares was sampled at 4 h intervals throughout an ovulatory cycle to determine cyclic FSH and LH patterns more accurately and to measure gonadotrophin pulse frequency during dioestrus. Synchronous pulses of FSH and LH occurred regularly in all mares between day 4 and day 12 (ovulation = day 0) with a mean (+/- SEM) frequency of 1.9 +/- 0.1 (FSH) or 1.6 +/- 0.1 (LH) pulses day-1. LH pulse amplitude declined (P < 0.0001) between day 4 and day 10, but FSH pulse amplitude remained large and stable, dipping slightly but not significantly on day 6. Daily mean FSH concentrations exceeded (P < 0.0001) early oestrous values between day 4 and day 5, and between day 7 and day 10. However, significantly different patterns were obtained when once-a-day sampling was simulated by selecting samples collected at 08:00 h or noon. LH was higher during the periovulatory surge than during dioestrus (P < 0.0001) and profiles were similar whether daily means or selected samples were used. It is concluded that: (1) the marked pulsatility of plasma FSH during dioestrus makes once-a-day sampling misleading for determining FSH profiles; (2) the dioestrous pattern of large, slow FSH pulses was consistent among mares, unlike that of the daily mean FSH profiles; and (3) no discrete FSH 'surges' were observed during dioestrus, although FSH pulse amplitude tended to undergo alternate increases and decreases. A period of higher amplitude FSH pulses preceded ovulation by 10.2 +/- 0.7 days, which corresponds to the approximate time the ovulatory follicle emerges. Therefore, it is possible that the signal for follicular recruitment in mares is intermittent excursions of plasma FSH above a threshold value. PMID- 9861173 TI - Enhancement of bovine oocyte fertilization in vitro with a bovine oviductal specific glycoprotein. AB - The purpose of this study was to determine the effect of a partially purified bovine oviductal glycoprotein (bOGP) on fertilization rates of bovine oocytes. The effect of albumin (control protein) or bOGP at 100 micrograms ml-1 during the 16-18 h fertilization period was evaluated in a standard IVF system using a sperm concentration between 0.5 and 0.125 x 10(6) spermatozoa ml-1. bOGP maintained a higher (P < 0.05) fertilization rate (62.0% versus 31.2%) at 0.125 x 10(6) spermatozoa ml-1 compared with the albumin control. The enhancement of fertilization by bOGP was blocked by the inclusion of a specific antibody to bOGP, whereas the antibody with albumin had no effect. A 2 h gamete preincubation step was subsequently included in the IVF procedure (0.125 x 10(6) spermatozoa ml 1) to determine whether the effect of bOGP was mediated through an interaction with the oocyte, the spermatozoon or both. When oocytes were preincubated with bOGP the fertilization rates were higher (P < 0.05) than with the albumin control (oocytes and spermatozoa exposed to albumin), whereas preincubation of spermatozoa with bOGP did not affect fertilization rates. There was no synergistic effect of preincubating oocytes and spermatozoa with bOGP. The increase in fertilization rate achieved by preincubating oocytes with bOGP was blocked with a specific antibody to bOGP. These results suggest that the increase in fertilization rates observed when bOGP is included during the 16-18 h fertilization period are primarily mediated through the interaction of bOGP with the oocyte since the same facilitatory effect was observed with a 2 h preincubation of oocytes before IVF. The ability to block these effects with a polyclonal antibody specifically generated against bOGP shows that this biological activity is due to bOGP. In summary, bOGP enhances fertilization in bovine oocytes whether it is included during preincubation or insemination and this appears to be due to a direct effect on the oocyte. PMID- 9861174 TI - Regulation of collagenolysis and cell death by plasmin within the formative stigma of preovulatory ovine follicles. AB - Collagen breakdown and apoptotic cell death within the apex of the preovulatory ovine follicle are characteristic of impending rupture. It has been hypothesized that plasmin regulates these two responses by activating collagenases and tumour necrosis factor alpha (TNF-alpha), respectively. Apical plasmin bioactivity, collagenolysis, TNF-alpha-mediated fragmentation of cellular DNA (a marker of apoptosis), stigma formation, and follicular rupture in sheep were inhibited by intrafollicular injection of alpha 2-antiplasmin. Explants of follicular wall released hydroxyproline-containing peptides (degraded collagen) and bioactive TNF alpha upon exposure to plasmin. These results indicate that plasmin has an essential intermediary role in the biomechanics of ovulatory ovarian tissue dissolution. PMID- 9861175 TI - Changes in circulating and ovarian concentrations of bioactive tumour necrosis factor alpha during the first ovulation at puberty in rats and in gonadotrophin treated immature rats. AB - Tumour necrosis factor alpha (TNF-alpha) concentrations were measured during periods of controlled and natural follicular development and ovulation in rat ovaries. Concentrations of bioactive TNF-alpha were determined in the ovaries and sera of rats during puberty (the period of vaginal opening and the first ovulation) and in immature rats after gonadotrophin treatment. Ovaries and sera were collected from 33-, 35-, 37-, 39-, 41- and 43-day-old rats (n = 6 or 7 per group); vaginal opening occurs on day 35. The presence of ovarian follicles and corpora lutea or ova in the oviducts was assessed. For gonadotrophin treatment, a single subcutaneous injection of 5 iu equine chorionic gonadotrophin (eCG) was administered at 08:00 h to 28-day-old rats to stimulate follicular development. A single subcutaneous injection of 10 iu human chorionic gonadotrophin (hCG) was administered 48 h later to induce ovulation. Ovaries and sera from three to six animals per group were collected 0, 3, 24, 48, 51, 54, 57, 60 and 72 h after injection of eCG. At puberty, ovarian concentrations of TNF-alpha were highest (approximately 1.1 fg micrograms-1 ovarian protein) before vaginal opening and the first ovulation. After vaginal opening and ovulation at day 37, ovarian concentrations of TNF-alpha were markedly reduced (0.091 fg microgram-1 ovarian protein) and remained low up to day 43. Serum concentrations of TNF-alpha remained low throughout the period of vaginal opening and the first ovulation (8 32 pg ml-1). In 43-day-old rats serum concentrations of TNF-alpha increased (105 pg ml-1). In the immature ovaries of 28-day-old rats TNF-alpha concentrations were highest before injection of eCG (approximately 1.2 fg micrograms-1 ovarian protein) and decreased to approximately 0.4 fg microgram-1 protein 3 h after injection. TNF-alpha concentrations decreased further 24 h after eCG injection (< 0.1 fg microgram-1 protein) and remained low until 48 h after eCG injection. Serum concentrations of TNF-alpha did not change during the 48 h period after injection of eCG. hCG was administered 48 h after eCG, and ovarian and serum TNF alpha concentrations increased transiently. Serum TNF-alpha concentrations increased 3 h after hCG and remained elevated until 9 h after injection, after which concentrations decreased. Ovarian concentrations of TNF-alpha increased 6 h after hCG, peaked (approximately 0.5 fg microgram-1 protein), and then declined. These results indicate that during puberty and the first ovulation, circulating and ovarian TNF-alpha concentrations change. In addition, exogenous gonadotrophins alter circulating and ovarian TNF-alpha concentrations. These data suggest that TNF-alpha has a role in follicular development and ovulation during puberty and in immature rats treated with gonadotrophins to induce ovulation. PMID- 9861176 TI - Nuclear transfer from sexed parent embryos in cattle: efficiency and birth of offspring. AB - The objectives of this study were to evaluate the efficiency and reliability of embryo sexing from isolated single blastomeres, and after nuclear transfer to examine the influence of the sex of donor embryos on development in vitro and in vivo up to calving. The sex of the donor embryo was determined by revealing a specific Y DNA sequence by PCR and electrophoresis after isolation of one, two, three, or more than five cells. The efficiency of sex determination was over 90% and reliability was 100% independent of the number of blastomeres used. In a second experiment, sex was determined from a single cell and the other cells were used for nuclear transfer. The effect of sex on in vitro development was studied in 386 male and 314 female reconstructed embryos derived from 19 male and 14 female parent embryos, respectively. Developmental competence in vitro of male and female constructs over 7 days was not statistically different (25.2 and 23.1% blastocysts on day 7, respectively; P > 0.05). After the transfer of predetermined male (n = 30) and female (n = 27) cloned embryos into recipient heifers, no effect of sex was observed on pregnancy rates at day 21, 35 and 90, or on calving rates (P > 0.05). These rates did not differ between single and twin transfer (P > 0.05). The sex of the calves born always corresponded to that determined from a single blastomere. These results show that sex can be determined accurately when using a single blastomere before nuclear transfer and that the sex of the parent embryo does not affect in vitro development or in vivo survival rates of cloned embryos. PMID- 9861177 TI - Spatio-temporal changes of prolyl 4-hydroxylase in granulosa cells during ovulation in eCG-hCG-treated immature rat ovaries. AB - The immunolocalization of prolyl 4-hydroxylase (PHase), a key enzyme of collagen synthesis, and the effects of anti-progesterone RU486 on PHase during the ovulatory process in eCG-hCG-treated immature rat ovaries were studied to investigate the mechanisms of tissue repair in follicle walls after follicular rupture. Immunolocalization of PHase was studied using an anti-rat PHase subunit monoclonal antibody, and the amount of immunoreactive PHase was measured by enzymeimmunoassay. No obvious immunolocalization of PHase was observed in theca cells throughout the ovulatory process except just after follicular rupture. In contrast, in granulosa cells, PHase was first observed at 9 h after the hCG injection, and the staining intensity apparently increased from 9 to 15 h, especially around the apex of preovulatory follicles and the orifice of ruptured follicles. Consistent with these observations, PHase concentration in granulosa cells isolated from the ovaries significantly increased by 9 h (0.45 +/- 0.03 pg per cell), and reached a peak at 15 h (0.66 +/- 0.06 pg per cell) after the hCG injection. This peak was inhibited when 20 mg RU486 kg-1 was administered at 8 h (0.46 +/- 0.05 pg per cell), and the RU486-inhibited PHase concentration was recovered by the concomitant administration of 10 mg progesterone kg-1 (0.65 +/- 0.02 pg per cell). The results suggest that PHase expressed in granulosa cells may play an important role in the repair of ruptured follicle walls, via progesterone-dependent PHase production. PMID- 9861178 TI - Correlation between the prevalence of certain fungi and sick building syndrome. AB - OBJECTIVE: To examine the role of fungi in the production of sick building syndrome. METHODS: A 22 month study in the United States of 48 schools (in which there had been concerns about health and indoor air quality (IAQ). Building indoor air and surface samples, as well as outdoor air samples were taken at all sites to look for the presence of fungi or their viable propagules. RESULTS: Five fungal genera were consistently found in the outdoor air and comprised over 95% of the outdoor fungi. These genera were Cladosporium (81.5%), Penicillium (5.2%), Chrysosporium (4.9%), Alternaria (2.8%), and Aspergillus (1.1%). At 20 schools, there were significantly more colony forming units per cubic metre (CFU/m3) (p < 0.0001) of propagules of Penicillium species in the air samples from complaint areas when compared with the outdoor air samples and the indoor air samples from noncomplaint areas. At five schools, there were more, although not significant (p = 0.10), Penicillium propagules in the air samples from complaint areas when compared with the outdoor air samples and the indoor air samples from noncomplaint areas. In 11 schools, the indoor air (complaint areas) fungal ratios were similar to that in the outdoor air. In these 11 schools Stachybotrys atra was isolated from swab samples of visible growth under wetted carpets, on wetted walls, or behind vinyl wall coverings. In the remaining 11 schools, the fungal ratios and CFU/m3 of air were not significantly different in different areas. Many of the schools took remedial action that resulted in an indoor air fungal profile that was similar to that outdoors. CONCLUSIONS: Propagules of Penicillium and Stachybotrys species may be associated with sick building syndrome. PMID- 9861179 TI - Survey of injuries among West End performers. AB - OBJECTIVES: To obtain more information about injuries of West End performers. METHODS: A retrospective survey of 269 performers appearing in 20 West End productions (12 dramas and eight musicals). RESULTS: In current productions, 46% of all performers sustained at least one injury for an average of 0.87 injuries per performer. Lower extremity injuries were the most common for dancers (52.2% of injuries) and actors (43.2%) with neck and back injuries the second most common. Sprains and strains were the most common diagnoses. 61% of performers thought that their injuries were preventable. Most performers consulted nonphysician healthcare providers. Factors significantly influencing the risk of injuries for performers include female sex, a history of previous injuries, missed performances due to previous injuries, more physically demanding roles, and performing on raked (angled) stages. CONCLUSION: West End performers commonly sustain injuries. Although primary prevention of most theatrical injuries is not possible, modification of raked stages may reduce the incidence. This study may be helpful to the growing number of healthcare providers who practice performing arts medicine and may stimulate additional concern and research in the medical and theatrical communities about the performance injuries of professionals, amateurs, and theatrical students worldwide. PMID- 9861180 TI - Cancer incidence in urban bus drivers and tramway employees: a retrospective cohort study. AB - OBJECTIVE: To investigate the risk of cancer associated with exposure to air pollution among bus drivers and tramway employees. METHODS: A retrospective cohort study of 18,174 bus drivers or tramway employees in Copenhagen in the period 1900-94. Data on employment were obtained from company files. Information on cancer was obtained from the Danish Cancer Registry. RESULTS: Findings showed that bus drivers or tramway employees had an increased risk of all malignant neoplasms (standardised incidence ratio (SIR) 1.24, 95% confidence interval (95% CI) 1.19 to 1.30). The relative risk was significantly increased for both men and women (SIR 1.24, 95% CI 1.19 to 1.30 and 1.28, 1.06 to 1.53, respectively). People employed for < 3 months had no increased risk of cancer (1.04, 0.81 to 1.31). For men who were employed for > 3 months the risk of lung cancer (1.6, 1.5 to 1.8), laryngeal cancer (1.4, 1.0 to 1.9), kidney cancer (1.6, 1.3 to 2.0), bladder cancer (1.4, 1.2 to 1.6), skin cancer (1.1, 1.0 to 1.2), pharyngeal cancer (1.9, 1.2 to 2.8), rectal cancer (1.2, 1.0 to 1.5) and liver cancer (1.6, 1.2 to 2.2) was significantly increased. For women employed for > 3 months the risk of lung cancer was significantly increased (2.6, 1.5 to 4.3). CONCLUSION: This cohort study shows that bus drivers and tramway employees are at an increased risk of developing several types of cancer. This might be due to the exposure to air pollution during working hours or to other risk factors, primarily smoking. PMID- 9861181 TI - Case-control study of occupational exposures and male breast cancer. AB - OBJECTIVE: To investigate whether risk of male breast cancer is associated with workplace exposures. METHODS: A case-control study of 178 cases of male breast cancer and 1041 controls was carried out with data from the United States national mortality follow-back survey, which collected questionnaire information from proxy respondents of a 1% sample of all 1986 United States deaths among subjects aged 25-74 years. Occupational exposure to electromagnetic fields, high temperatures, polycyclic aromatic hydrocarbons (PAHs), herbicides, other pesticides, and organic solvents was assessed by applying job-exposure matrices, based on the 1980 United States census occupation and industry codes, to the longest job held by study subjects as reported by the informants. A socioeconomic status index was created by combining information on annual family income, education, assets, and occupation to assess the association of socioeconomic status with male breast cancer. Relative risks were derived from logistic regression modelling, which included age, socioeconomic status, marital status, and body mass index, as well as occupational exposures. RESULTS: Risk for male breast cancer increased significantly with increasing socioeconomic status index (test for trend: p < 0.01), but the risks associated with individual socioeconomic status variables were smaller and the trends were not significant. A significant increase in risk of male breast cancer was associated with employment in blast furnaces, steel works, and rolling mills (odds ratio (OR) 3.4; 95% confidence interval (95% CI) 1.1 to 10.1, based on six cases), and motor vehicle manufacturing (OR 3.1; 95% CI 1.2 to 8.2, based on seven cases). However, exposures to electromagnetic fields, high temperature, PAHs, herbicides, other pesticides, and organic solvents were not associated with risk of male breast cancer. CONCLUSIONS: The role of workplace exposures in increasing risk of breast cancer among men employed in motor vehicle manufacturing and in blast furnaces, steel works, and rolling mills deserves further investigation. The finding on socioeconomic status suggests that, as well as reproductive factors, other lifestyle factors such as diet that may be related to high socioeconomic status in men should be investigated further. PMID- 9861182 TI - Air pollution and daily mortality in Rome, Italy. AB - OBJECTIVES: To assess the relation between several daily indicators of air pollution (particulates and gases) and daily mortality in the metropolitan area of Rome and in the central part of the city. METHODS: Time series analysis. The associations between daily concentrations of pollutants (particles, SO2, NO2, CO, O3) recorded by five fixed monitors and daily total mortality in the period from January 1992 to June 1995 were evaluated. The analysis included examination of the pollution effect on mortality by place of residence within the metropolitan area, by season, age, place of death (in and out a hospital), and cause of death (cardiovascular and respiratory disease). The Poisson model included loses smooth functions of the day of study, mean temperature, mean humidity, and indicator variables for day of the week and holidays. RESULTS: The mean daily number of deaths was 56.9 (44.8 among people > or = 65 years old). A mean of 36.3 deaths occurred in the city centre; 37.3 deaths a day were recorded in a hospital. Total mortality was significantly associated with a 10 micrograms/m3 increase in particles (0.4%) on that day (log 0), and with a 10 micrograms/m3 increase in NO2 at lag 1 (0.3%) and lag 2 (0.4%) (1 and 2 days before, respectively). The effect of particles (lag 0) and of NO2 (lag 2) on total mortality was higher among those living in the city centre (0.7% and 0.5%, respectively). The risk estimates were higher in the warmer season (1.0% and 1.1%, respectively), whereas no difference was found for those dying in or out of the hospital. The effect of particles was robust to a sensitivity analysis and to the inclusion of NO2 in the regression model. CONCLUSIONS: Increase in particulates and NO2, generated by the same mobile combustion sources, is associated with a short term increase in mortality in Rome. The effect is more evident among residents in the city centre, where the levels of exposure to pollutants recorded by fixed monitors are probably more reliable indicators of personal exposure. PMID- 9861183 TI - Small area study of mortality among people living near multiple sources of air pollution. AB - OBJECTIVE: In the area of Malagrotta, a suburb of Rome (Italy), a large waste disposal site, a waste incinerator plant, and an oil refinery plant became operational in the early 1960s and have represented three major sources of air pollution. To evaluate the potential health risk due to airborne contamination around these point sources, a small area analysis of mortality was conducted. Cancer of the liver, larynx, lung, kidney, lymphatic, and haematopoietic systems were evaluated. METHODS: Sex and age specific mortality (1987-93) and population denominators (1991) were available for the census tracts of the metropolitan area of Rome. Standardised mortality ratios (SMRs) were computed separately for males and females in bands of increasing distance from the plants, up to a radius of 10 km. Stone's test for the decline in risk with distance was performed with increments in radius of 1 km; SMRs were also computed after adjusting for a four level index of socioeconomic status. RESULTS: No overall excess or decline in risk with distance was found for liver, lung, and lymphohaematopoietic cancers in either sex. For laryngeal cancer, an increased but not significant risk was found at 0-3 km and at 3-8 km. A significant decline with distance in mortality from laryngeal cancer was found among men (p = 0.03); the trend remained after adjusting for the socioeconomic index (p = 0.06). CONCLUSIONS: The study showed no association between proximity to the industrial sites and mortality for most of the several conditions considered. However, mortality from laryngeal cancer declined with distance from the sources of pollution. This result is interesting, as previous findings of an increased risk of laryngeal cancer near incinerators have been controversial. PMID- 9861184 TI - Health survey of former workers in a Norwegian coke plant: Part. 1. Estimation of historical exposures. AB - OBJECTIVE: To estimate historical exposure levels at a coke plant for all agents considered to be of importance for epidemiological studies of mortality and cancer incidence. METHODS: Time weighted average exposure (8 h TWA) was estimated based on personal measurements for polycyclic aromatic hydrocarbons (PAHs) and carbonaceous particulates. Exposure to quartz was estimated relative to the concentration of carbonaceous particulates. These estimates were adjusted for the use of airstream helmets. Exposure to other agents were estimated qualitatively (asbestos, benzene, and arsenic) or semi-quantitatively (carbon monoxide (CO) and heat) based on measurements and other indicators of exposure. RESULTS: Exposure to PAHs was highest for those who worked at the top of the ovens (300 micrograms/m3) in the period from 1970-6. The estimated PAH exposure was reduced to an average of 65 micrograms/m3 after the introduction of exposure control measures in 1976. The estimates for carbonaceous particulates ranged from 1 to 16 mg/m3, with the highest exposure for workers at the top of the ovens and at the coke screening station. CONCLUSIONS: The exposure of greatest concern in this study is to PAHs, but exposures to carbonaceous particulates and CO may also be of importance. The major limitations of this study are the lack of personal measurements before 1975 and the total lack of measurements for some of the exposed categories of workers. Despite these limitations, we think that this assessment reflects the actual exposures for most of the former employees. The assessment thus provides a reasonable tool for the subsequent epidemiological study and for future epidemiological follow up studies at the coke plant. PMID- 9861185 TI - Health survey of former workers in a Norwegian coke plant: Part 2. Cancer incidence and cause specific mortality. AB - OBJECTIVES: A Norwegian coke plant that operated from 1964 to 1988 was investigated to ascertain whether the male workers in this plant had increased morbidities of cancer or increased mortality from specific causes, particularly associated with specific exposures at the coke plant. METHODS: Personal data on all the employees of the plant were obtained from the plant's archives. With additional data from the Norwegian Bureau of Statistics we identified 888 male former workers at the plant. Causes of death were obtained from the Norwegian Bureau of Statistics, and cancer diagnoses from the Norwegian Cancer Registry. The results were compared with national averages adjusted for age. Specific exposures were estimated with records of actual measurements done at the plant and interviews with former workers at the plant. RESULTS: A significant excess of stomach cancer (standardised incidence ratio (SIR) 2.22, 95% confidence interval (95% CI) 1.01 to 4.21) was found. Mortality from ischaemic heart disease and sudden death was positively associated with work in areas which entailed peak exposures to CO. When considering work in such areas the past 3 years before death, the association was significant (p = 0.01). The last result is based on only two deaths. CONCLUSIONS: Considering the short follow up time and the small size of the cohort the results should be interpreted with a certain caution. The positive results would justify a re-examination of the cohort at a later date. PMID- 9861186 TI - Occupational exposures to solvents and aluminium and estimated risk of Alzheimer's disease. AB - OBJECTIVES: To study the role of occupational exposures to solvents and aluminium in the aetiology of Alzheimer's disease (AD). An industrial hygienist rated exposure. METHODS: 89 subjects diagnosed with probable AD were matched by age, sex, and type of informant to 89 controls. Subjects were identified from a large health maintenance organisation in Seattle, WA. A complete occupational history was obtained from spouses of cases and controls as well as from controls themselves. After the interview an industrial hygienist, blinded to case-control status, rated exposures. RESULTS: Non-significant associations were found between AD and ever having been occupationally exposed to solvents (odds ratio (OR) 1.77, 95% confidence interval (95% CI) 0.81 to 3.90) and aluminium (OR 1.46, 95% CI 0.62 to 3.42). Although an increasing risk was found with increasing number of years of exposure to solvents, there was an inverse association between exposure intensity and AD, and measures of cumulative exposure taking into account both intensity and duration of exposure were not significant. Analysis of the age at which half the cumulative exposure to solvents was achieved showed that an older age incurred a greater risk of AD than a younger age. However, the total amount of exposure carried no risk. CONCLUSIONS: The results suggest that lifetime occupational exposure to solvents and aluminium are not likely to be important risk factors for Alzheimer's disease. PMID- 9861187 TI - Best estimate of the magnitude of mortality due to occupational exposure to hazardous substances. AB - OBJECTIVES: With a proportionate attributable risk approach, to estimate the magnitude of premature mortality induced by exposure to hazardous substances in the Australian workforce. METHODS: Estimates of the magnitude of mortality induced by exposure to hazardous substances in the Australian work-force were calculated by the proportionate attributable risk approach after careful consideration of options for methodological refinements. The main data sources used were unit mortality datasets (individual deidentified death records), and tabulations when these were unavailable. RESULTS: The estimated number of deaths that occurred in Australia each year from occupational exposure to hazardous substances was 2290; 78% of these were men. The rate of mortality attributable to occupational exposure to hazardous substances was three to four times greater in male workers than in female workers. Male (productive) person-years of life lost (PYLL) were generally eight times higher than female PYLL. Cancer was the prime cause of death, followed by renal, cardiovascular, neurological, and chronic respiratory disease. Acute toxic episodes accounted for a small proportion of mortality but yielded a much larger proportion of PYLL, reflecting the relatively young ages of those who experienced fatal effects. CONCLUSIONS: Although national estimates of the proportions of mortality attributable to occupational exposure to hazardous substances seemed to be validly derived, uncertainties remain associated with the lack of an empirical basis for derivation of proportionate risk fractions used in the calculations. The finding of an appreciable burden of mortality that is attributable to past occupational exposure to hazardous substances emphasises the necessity for occupational health and safety authorities to measure and reduce worksite exposures. There is also an incentive to encourage the construction of appropriately designed cohort studies across industries and occupational groupings so that, ultimately, risk estimates may be directly calculated and applied to total workforce data for the definitive estimation of the magnitude of harm induced by occupational exposure to hazardous substances. PMID- 9861188 TI - Myocardial infarction in male and female dominated occupations. AB - The aim of the study was to investigate whether workers in jobs dominated by the opposite sex have an increased risk of myocardial infarction (MI). A case referent study was carried out to estimate the relative risk of first MI in different occupational groups. The study base comprised all men and women in five counties in the middle of Sweden during 1976-84. Cases of MI were identified from both hospital discharge records and death records. Information on occupation was obtained from two consecutive censuses. Primary health related selection was analysed for men with data from the physical examination of conscripts to compulsory military service in 1969-70 combined with data from the censuses of 1970-90 and data on early retirement in 1971-92. Increased risk of MI was found among both women (relative risk (RR) 1.41, 95% confidence interval (95% CI) 1.15 to 1.73) and men (1.21, 1.10 to 1.32) in blue collar jobs where men predominate, and among men with white collar jobs (1.26, 1.09 to 1.45) where women predominate. However, the increased risk among men in white collar jobs was probably due to negative health selection into these occupations. These results do not support the notion that being of the sexual minority in an occupation is in itself an important risk factor for MI. PMID- 9861189 TI - Exposure to asphalt or bitumen fume and renal disease. PMID- 9861190 TI - Cancer risk in the rubber industry: a review of recent epidemiological evidence. PMID- 9861191 TI - Inhalation of ammonium nitrate fuel oil explosive (ANFO): and possible concomitant exposure. PMID- 9861192 TI - Occupational asthma due to amylase. PMID- 9861193 TI - Health of children born to medical radiographers. PMID- 9861194 TI - Platinum analogues in the treatment of recurrent high grade astrocytoma. PMID- 9861195 TI - Clinical implications of molecular abnormalities in lung cancer. PMID- 9861196 TI - Platinum-DNA adduct, nucleotide excision repair and platinum based anti-cancer chemotherapy. AB - Clinical studies performed by several groups suggest that platinum-DNA adduct- measured in malignant or non-malignant cells from cancer patients--may be an important marker for clinical biological effect of platinum-based chemotherapy. DNA repair is clearly an important effector of resistance to platinum-based DNA damaging agents in tissue culture, although its role in effecting clinical resistance to these agents is not completely clear. In recent years, it has become apparent that DNA repair is an extremely complex process. Processes within DNA repair that may contribute to one or more drug resistance phenotypes include 0-6-alkytransferase activity, base excision repair, mismatch repair, nucleotide excision repair (NER), and gene specific repair. Clearly, several of these processes may concurrently show increased activity within any single cell, or tumor, at any one time. For platinum compounds, in vitro data clearly show that NER is the DNA repair pathway responsible for the repair of cisplatin-DNA damage. One critical gene within NER is ERCC1. Data exist in human ovarian cancer and in human gastric cancer that ERCC1 may be a useful marker for clinical drug resistance when platinum-based systemic chemotherapy is utilized. Although the data suggest that the relative ERCC1 mRNA level may be a good marker for NER activity in human ovarian cancer, it is unclear whether expression of this gene has any relationship to other pathways of DNA repair. PMID- 9861197 TI - In-vitro metabolism of anti-cancer drugs, methods and applications: paclitaxel, docetaxel, tamoxifen and ifosfamide. PMID- 9861198 TI - Resident's project on osteoporosis flies with Shuttle. PMID- 9861199 TI - UBC gets to heart of problem. PMID- 9861200 TI - The evidence for insulin lispro. PMID- 9861201 TI - Assessing osteoporosis risk. PMID- 9861202 TI - Snowbirds: an unwelcome sign that winter's coming. PMID- 9861203 TI - "Yes" to exercise for breast cancer survivors. PMID- 9861204 TI - Participation in clinical trials among women living with HIV in Canada. Canadian Women's HIV Study Group. AB - BACKGROUND: To describe participation in clinical trials among HIV-positive women enrolled since 1993 in the Canadian Women's HIV Study, a prospective open cohort study. METHODS: All HIV-positive women being followed at hospital-based or community-based clinics at 28 sites in 11 Canadian cities were eligible to participate in the Canadian Women's HIV Study. Baseline and follow-up information was collected for 413 women every 6 months by study nurses using standardized questionnaires. Data included sociodemographic variables, HIV exposure group, CD4 count, disease classification, use of antiretroviral therapies and participation in clinical trials. RESULTS: At study intake 15.0% (62/413) of the women had participated in a clinical trial; an additional 8.5% (35/413) participated during a median follow-up of 18 months. Multivariate analysis revealed that the following factors were independently associated with participation in a clinical trial: white race (adjusted odds ratio [OR] 3.38, p = 0.001), current use of antiretroviral therapy (adjusted OR 2.01, p = 0.008), completion of secondary school (adjusted OR 1.97, p = 0.024) and residence in the Prairies or Atlantic provinces (adjusted OR 1.98, p = 0.043). INTERPRETATION: Although the overall clinical trial participation rate of 23.5% was relatively high among HIV-positive women, injection drug users were underrepresented in this study population, and non-white women, women who did not complete high school and women not receiving antiretroviral therapy were less likely than white women, women of higher education and women receiving antiretroviral therapy to participate in clinical trials in Canada. Because of the importance of trial participants being representative of the population for which therapeutic agents are intended, HIV clinical trials must recruit women with lower literacy levels, non-white women, women not receiving antiretroviral therapy and women who are injection drug users to ensure generalizability of research findings. Further study is needed to assess factors that act as barriers and motivators to women's participation in HIV clinical trials. PMID- 9861205 TI - Screening for prostate cancer: estimating the magnitude of overdetection. AB - BACKGROUND: No randomized controlled trial of prostate cancer screening has been reported and none is likely to be completed in the near future. In the absence of direct evidence, the decision to screen must therefore be based on estimates of benefits and risks. The main risk of screening is overdetection--the detection of cancer that, if left untreated, would not cause death. In this study the authors estimate the level of overdetection that might result from annual screening of men aged 50-70. METHODS: The annual rate of lethal screen-detectable cancer (detectable cancer that would prove fatal before age 85 if left untreated) was calculated from the observed prostate cancer mortality rate in Quebec; the annual rate of all cases of screen-detectable prostate cancer was calculated from 2 recent screening studies. RESULTS: The annual rate of lethal screen-detectable prostate cancer was estimated to be 1.3 per 1000 men. The annual rate of all cases of screen-detectable prostate cancer was estimated to be 8.0 per 1000 men. The estimated case-fatality rate among men up to 85 years of age was 16% (1.3/8.0) (sensitivity analysis 13% to 22%). INTERPRETATION: Of every 100 men with screen-detected prostate cancer, only 16 on average (13 to 22) could have their lives extended by surgery, since the prostate cancer would not cause death before age 85 in the remaining 84 (78 to 87). PMID- 9861206 TI - The evolution of clinical trials: inclusion and representation. PMID- 9861207 TI - Prostate cancer screening: waiting for Godot. PMID- 9861208 TI - Health services research and personal health information: privacy concerns, new legislation and beyond. PMID- 9861209 TI - Prostate cancer: 7. Radiation therapy for localized disease. PMID- 9861210 TI - The early years of radiation protection: a tribute to Madame Curie. PMID- 9861211 TI - As blue as Lake Louise. PMID- 9861213 TI - US editorial writers put Canadian health care under microscope. AB - Editorial writers from the US descended on Ottawa recently for their annual meeting, and CMAJ contributing editor Charlotte Gray was one of the speakers. She said the visitors received widely differing views on the Canadian health care system and may have emerged from the meeting more confused than informed. PMID- 9861212 TI - Ciguatera fish poisoning. PMID- 9861215 TI - Hospital mergers can bring breath of fresh air, CEO says. Interview by Heather Kent. PMID- 9861214 TI - Ontario's attempt at primary care reform hits another snag. AB - Ontario is the latest province to step into the health care reform spotlight. Proponents hope its proposed series of primary care pilot projects, built around the concept of patient rostering, will improve the delivery of care. If the project goes ahead, doctors in 5 participating sites will be paid under a capitation system, with payments starting at $70.29 annually for men aged between 25 and 35. PMID- 9861216 TI - The use and abuse of restraints. PMID- 9861217 TI - Africa's population problems not limited to Africa. PMID- 9861218 TI - Planning motherhood. PMID- 9861219 TI - Informing women about folic acid. PMID- 9861220 TI - Asthenia and paralysis. PMID- 9861222 TI - Women's decisions about hormone replacement therapy after education and bone densitometry. AB - BACKGROUND: The decisions that postmenopausal women make about whether to start hormone replacement therapy may depend on the potential risks and benefits of such therapy as well as their risk for osteoporosis-related fractures. This study examined the decisions made by women at risk for osteoporosis-related fractures who were educated about hormone replacement therapy and who were given information about their bone mineral density. METHODS: The study employed a prospective cohort design. Thirty-seven post--menopausal women with risk factors for osteoporosis-related fractures were recruited from an orthopedic clinic at a teaching hospital in Hamilton, Ont. The women were given an education kit (consisting of an audio tape and a work-book) to clarify the benefits and risks of hormone replacement therapy. Two to 4 weeks later, densitometry of the hip and the lumbar spine was performed. A summary of the risks, the densitometry findings and decisions about hormone replacement therapy were given to the women's family physicians for follow-up. Outcome measures included decisions about hormone replacement therapy, as well as use of such therapy and other medications at 12 months. RESULTS: After the education component alone, 10 (27%) of the women requested hormone replacement therapy. After densitometry testing, 4 more requested hormone replacement therapy (for a total of 14 women [38%]). At 12 months, 2 (5%) of the women had been lost to follow-up. Of the remaining 35, 6 (17%) were receiving hormone replacement therapy, 7 (20%) were using bisphosphonates, and 24 (68%) were taking calcium supplements. INTERPRETATION: These preliminary findings suggest that the combination of education about hormone therapy and feedback about bone density is associated with an increase in the use of hormone replacement therapy and other preventive medications by women at risk for osteoporosis-related fractures. However, the observed increase was small and so the clinical significance must be confirmed and clarified. PMID- 9861221 TI - Adverse effects associated with selective serotonin reuptake inhibitors and tricyclic antidepressants: a meta-analysis. AB - BACKGROUND: The use of antidepressant medications and the resulting costs have increased dramatically in recent years, partly because of the introduction of selective serotonin reuptake inhibitors (SSRIs). An assessment of the clinical and economic aspects of SSRIs compared with the older tricyclic antidepressants (TCAs) was initiated to generate information for purchasers of these drugs as well as clinicians. One component of this study was an examination of the adverse effects associated with the use of these drugs. METHODS: Searches of bibliographic databases (for January 1980 through May 1996) and manual scanning of both peer-reviewed publications and other documents were used to identify double-blind, randomized controlled trials involving at least one SSRI and one TCA. For the study of adverse effects, only trials that had at least 20 patients in each trial arm and that reported rates of adverse effects in both arms were retained. In total 84 trials reporting on 18 adverse effects were available. Meta analyses were undertaken to calculate pooled differences in rates of adverse effects. The question of whether the method of eliciting information from patients about adverse effects made a difference in the findings was also examined. Finally, differences in drop-out rates due to adverse effects were calculated. RESULTS: The crude rates of occurrence of adverse effects ranged from 4% (palpitations) to 26% (nausea) for SSRIs and from 4% (diarrhea) to 27% (dry mouth) for TCAs. The differences in the rates of adverse effects between the 2 types of drugs ranged from 14% more with SSRIs (for nausea) to 11% more with TCAs (for constipation). The results did not depend on the method of eliciting information from patients. There were no statistically significant differences between drug classes in terms of drop-outs due to adverse effects. INTERPRETATION: SSRIs and TCAs are both associated with adverse effects, although the key effects differ between the drug classes. Further explanation of the adverse effects and their relation to discontinuation of medication will require better studies involving prospective collection of quality-of-life data. PMID- 9861223 TI - Osteoporosis and bone densitometry: does the emperor have clothes? PMID- 9861224 TI - Prostate cancer: 6. Surgical treatment of localized disease. AB - A 65-year-old man undergoes a routine checkup before retiring. His wife has urged him to have his prostate examined, because she has read about testing for prostate cancer and a friend has just died of this disease. During the rectal examination, the man's physician discovers some firmness in the right lobe of the prostate gland. The patient has had no urinary symptoms and is in excellent general health. Sexual function is normal. There is no history of prostate cancer; his father died of a stroke at age 86 years. Testing shows that the patient's prostate-specific antigen level is 9.3 ng/mL, and he is referred to a urologist. Transrectal ultrasound-guided needle biopsy reveals adenocarcinoma with a Gleason score of 7 (intermediate grade). At a follow-up meeting with his physician, the patient says, "I have been doing some research, and it appears that I should have treatment. However, what is less clear to me is what form of therapy is best--surgery or radiation treatment. Please tell me what you can about the state of the art with respect to surgery." PMID- 9861225 TI - Enzyme replacement therapy for Gaucher's disease: the early Canadian experience. AB - BACKGROUND: The management of severe Gaucher's disease was dramatically improved by the development of enzyme replacement therapy. However, this treatment is very costly (currently about $21,000 per infusion for adults at the starting dose recommended by the manufacturer). The goal of this study was to determine how enzyme replacement therapy was being prescribed and financially supported in various parts of Canada. In addition, demographic and outcome information was elicited. METHODS: Prescribing physicians were identified through professional associations and with the help of the manufacturer of the enzyme preparations used for the treatment of Gaucher's disease. The physicians were surveyed by questionnaire in July 1995. The study included all patients in Canada who had received enzyme replacement therapy for Gaucher's disease before July 1, 1995. RESULTS: A total of 25 patients (15 children and 10 adults) with type 1 Gaucher's disease, the common nonneuronopathic variant of the disease, were receiving enzyme replacement therapy by the end of 1995. The indications for treatment included massive splenomegaly, growth failure, and severe bony, hematologic and pulmonary complications of the disease; no patients with mild disease were receiving treatment. Treatment regimens varied markedly (from 12 to 160 units of enzyme/kg per month). All the patients were reported to have responded well to therapy, based on serial measurements of hematologic indices, liver and spleen volumes, and numbers of bony crises as well as patients' subjective impressions. Financial support for therapy varied markedly from one province to another. None of the reporting physicians was aware of any patients with severe Gaucher's disease who were denied therapy as a result of inability to pay for the medication. Various agencies provided financial support for therapy, including both federal and provincial governments, private insurance carriers and the commercial supplier of the enzyme. In Ontario provincial health care officials accepted the development, by a multidisciplinary panel of medical experts, of formal guidelines for determining eligibility, on the basis of objective medical criteria, for reimbursement for enzyme replacement treatment. INTERPRETATION: Although some differences were found across the country with respect to the details of treatment, the indications for enzyme replacement therapy and the selection of severely affected patients were similar in the various provinces. However, financial support was inconsistent and varied among provinces and patients. This will prove to be a challenge in future, not only with respect to this disease but also for other diseases for which effective, expensive therapy has been developed. PMID- 9861226 TI - Reversible impairment of renal function associated with enalapril in a diabetic patient. AB - Acute renal failure and hyperkalemia due to angiotensin-converting enzyme inhibitors have been described in diabetic patients with other predisposing conditions. The case reported here involves a patient with type 1 diabetes mellitus, microalbuminuria and normal renal function who was treated with enalapril. Two years after initiation of this therapy, at a time when glycemic control was poor, he presented with symptomatic hyperkalemia and impaired renal function accompanied by hyporeninemic hypoaldosteronism. This case illustrates that reversible impairment of renal function and hyperkalemia can present after 2 years of treatment with angiotensin-converting enzyme inhibitors in patients with precipitating factors. PMID- 9861227 TI - Twelve deaths in Winnipeg: judge must ponder 48,000 pages of inquest testimony. AB - The Winnipeg inquest into the deaths of 12 young heart patients has ended after nearly 3 years of testimony, much of which centred on a surgeon's competence. The recommendations emerging from it are expected early next year. PMID- 9861228 TI - Royal College debates whether MDs should promote moderate consumption of alcohol. AB - Emerging news about the potential beneficial effects of moderate alcohol consumption raises some interesting challenges for physicians, who often come face to face with problems created by alcohol. Physicians on the affirmative side won a debate on the pros and cons of moderate drinking that was held during this fall's Royal College meeting. Pam Harrison explains how they did it. PMID- 9861229 TI - [New perspectives for Alzheimer patients: acetylcholine inhibition with Rivastigmine. Congress of the European Neurologic Society. Nice, 6 July 199]. PMID- 9861230 TI - MD reviewers' role in the new anticancer drug approval process in the newly established Japanese regulatory agency, PMDEC (Pharmaceuticals and Medical Devices Evaluation Center) PMID- 9861231 TI - Cytochrome P4501A1 and glutathione S-transferase M1 genotypes as risk factors for prostate cancer in Japan. AB - BACKGROUND: The p53 mutation spectrum of prostate cancers developing in Japan indicates a role for environmental factors. This suggests there might be differences in susceptibility due to genetic polymorphisms in metabolic activation enzyme genes. We analyzed genetic polymorphisms of the xenobiotic metabolizing enzymes, CYP1A1 and GSTM1. METHOD: Genotyping of CYP1A1 and GSTM1 was investigated by using allele-specific PCR in 115 prostate cancer (PCa) patients and 204 control patients. RESULTS: The CYP1A1 Val/Val genotype significantly increased the risk for PCa (OR = 2.6; 95% CI = 1.11-6.25) and the Ile/Val genotype showed a similar tendency (OR = 1.4; CI = 0.86-2.29). Individuals with the GSTM1 (0/0) genotype demonstrated a slightly increased risk (OR = 1.3; CI = 0.82-2.04). The combination of the CYP1A1 Val allele and GSTM1 (0/0) genotype was associated with a higher risk (OR = 2.3; CI = 1.18-4.48) than the CYP1A1 Val allele alone. When cases were analyzed by age at initial diagnosis, the relative risks with both the CYP1A1 Val allele and the GSTM1 (0/0) genotype were higher in the young group than in the old group (CYP1A1; OR = 1.7, CI = 0.89-3.17: GSTM1; OR = 1.6, CI = 0.84-2.99). The frequency of the GSTM1 (0/0) genotype was also higher in patients with advanced stage disease. In stage D, the OR was 1.7 with a CI of 0.93-3.17 and in stages A and B, the OR was 0.8 with a CI of 0.40-1.62. CONCLUSIONS: These results suggest that CYP1A1 and GSTM1 polymorphisms are linked to a propensity for PCa development. PMID- 9861232 TI - Significance of free to total PSA ratio in men with slightly elevated serum PSA levels: a cooperative study. AB - BACKGROUND: The ratio of free PSA in total PSA (f/t) has been reported to improve the diagnostic accuracy of prostate cancer in the group with slightly elevated serum PSA values. In Japanese cases, the clinical significance of f/t is still controversial. METHODS: The diagnostic significance of f/t in serum for prostate cancer was evaluated in a cooperative study. A total of 77 cases with prostate cancer and 224 with non-prostate cancer showing less than 20 ng/ml of total PSA were evaluated. RESULTS: Serum total and free PSA values were not affected by storage at 25 degrees C for 2 days. The determination of f/t was useful in the cases with a serum total PSA of 5.1-10 ng/ml; the specificity was 60% with a sensitivity of 90% at an f/t of 0.148. The positive predictive value for diagnosis of prostate cancer also increased to 54% from 34% of that in total PSA alone. In the range of 4.1-10 ng/ml, the cut-off value of f/t was 0.155 for obtaining relatively high specificity; sensitivity was 85% and specificity was 56.5%. CONCLUSIONS: Thus, the determination of f/t was considered to be an effective tool for discriminating the non-prostate cancer cases from those of prostate cancer. PMID- 9861233 TI - Results of transrectal ultrasound-guided biopsies and clinical significance of Japanese prostate cancer. AB - BACKGROUND: We review the outcomes of ultrasound-guided biopsy in consecutive patients and assess clinical significance of Japanese prostate cancer. METHODS: Examination was made of 1469 patients subsequent to transrectal ultrasound-guided biopsy of the prostate gland. For 84 patients, two or more sets of ultrasound guided biopsies were conducted following the initial negative results during this period. Two hundred and thirty-two patients with benign histology at the initial biopsy underwent transurethral resection of the prostate (TURP). The clinical significance of the cancers was assessed based on patient age and calculated tumor volume at diagnosis, assumed cancer volume doubling time and life expectancy in the Japanese male population. RESULTS: Overall, 327 of the 1469 patients (22.3%) had prostate carcinoma. Positive biopsy rates in patients with PSA 2.0 ng/ml or lower, 2.1-4.0 ng/ml, 4.1-10.0 ng/ml and 10.1 ng/ml or greater were 4.6%, 8.6%, 15.8% and 59.5%, respectively. Of the 232 patients who underwent TURP, 15 (6.5%) had cancer. Of the 84 patients subjected to the multiple sets of biopsies, 19 (22.6%) cancers were detected. Of the 203 cancers without distant metastasis at initial biopsy, 13.3%, 25.1%, 32.5% and 40.4% of tumors for 2-, 3-, 4- and 6-year tumor doubling times gave no indication of clinical significance. Nearly half these patients (43-52%) had clinical stage T1c disease. The estimated proportion of clinically insignificant tumors in repeat biopsy was virtually the same as first set biopsies. CONCLUSIONS: Low PSA was not necessarily an indication of indolent cancer and repeat biopsy did not often demonstrate clinically unimportant cancers. Many patients with stage T1c disease may eventually prove to require no treatment. PMID- 9861235 TI - Development of data-based semi-quantitative food frequency questionnaire for dietary studies in middle-aged Japanese. AB - BACKGROUND: We designed a data-based semi-quantitative food frequency questionnaire to clarify the relationship between food intake and lifestyle related diseases among middle-aged Japanese. METHODS: A total of 351 middle-aged individuals were recruited to a one-day weighed diet record survey in 1994. In all, 586 foods were consumed. Intake of 31 nutrients including energy, protein, fat, carbohydrate, vitamins, minerals and dietary fiber by food was computed by multiplying the weight of food consumed by its nutrient content. First, 252 foods with up to 90 cumulative % contribution to nutrient intake were selected. Of these, foods having apparently the same/similar nutrient content were combined into 206 foods by research dietitians. Next, 183 foods with up to 0.90 cumulative multiple regression coefficient and 90 cumulative % contribution were chosen. At this stage an additional food grouping was made. RESULTS: Finally, 102 foods/recipes were included in the questionnaire: rice (2 items), bread and noodles (11), eggs, milk and dairy products (10), soybean, soybean products and other beans (7), meat including beef, pork and chicken (12), fish (5), other fish, shellfish and fish products (10), green-yellow vegetables (8), other vegetables and mushrooms (7), edible roots (2), seaweeds (3), seeds (2), fruits (8), beverages (7) and confectioneries (8). The frequencies were classified into eight categories. Portion size was calculated for the respective foods largely from the one-day weighed diet record. CONCLUSIONS: The developed semi quantitative food frequency questionnaire substantially covered the intake of 31 nutrients and may be competent to rank middle-aged Japanese efficiently. PMID- 9861234 TI - Clinicopathological factors predicting retroperitoneal lymph node metastasis and survival in endometrial cancer. AB - BACKGROUND: By clarifying the significance of clinicopathological factors for retroperitoneal lymph node metastasis and survival of patients with endometrial cancer, we suggest ideas for optimal treatment of this disease. METHODS: A retrospective study was conducted in 310 women with endometrial cancer who underwent surgery with retroperitoneal lymphadenectomy. To evaluate retroperitoneal lymph node metastasis, age-adjusted and multivariable analyses were carried out for six clinicopathological factors including pathological grade, myometrial invasion, cervical invasion, peritoneal cytology, lymphatic permeation and vascular invasion. To evaluate survival, besides the above factors, a positive rate of metastasis of dissected retroperitoneal lymph nodes was included. RESULTS: In 40 patients (13%) with nodal metastasis, the average positive rate of metastasis of dissected retroperitoneal lymph nodes was 22%. For retroperitoneal lymph node metastasis, the odds ratio of deep myometrial invasion, cervical invasion and severe lymphatic permeation were 5.97, 2.72 and 12.01, respectively. For survival, the hazard ratios of the positive rates of metastasis of dissected retroperitoneal lymph nodes (both 25% and < 25%), positive peritoneal cytology and poor pathological grade were 7.10, 3.24, 3.82 and 3.27, respectively, and 5-year survival rates for them were 0, 50, 72 and 77%, respectively. CONCLUSIONS: For retroperitoneal lymph node metastasis, lymphatic permeation, deep myometrial invasion and cervical invasion were the independent prognostic factors. For survival, retroperitoneal lymph metastasis, poor pathological grades and positive peritoneal cytology were the independent prognostic factors. The positive rate of metastasis of dissected retroperitoneal lymph metastasis plays an important role in predicting survival of endometrial cancer. Lymph node biopsy is insufficient in treatment of this disease. PMID- 9861236 TI - Acute myelogenous leukemia with monosomy 7, inv(3) (q21q26), involving activated EVI 1 gene occurring after a complete remission of lymphoblastic lymphoma: a case report. AB - A 42-year-old female with a mediastinal tumor and massive pleural effusion ws admitted to our hospital in June 1993. Biopsy revealed lymphoblastic lymphoma. She had no evidence of distant metastasis except pleural effusion. Bone marrow examination revealed a normal karyotype (46, XY). The patient had been progression-free for more than 1 year after achieving complete remission by induction, consolidation and maintenance therapy according to the standard chemotherapy and involved-field radiation for lymphoblastic lymphoma. From May 1996 progressive leukopenia and thrombocytopenia developed. The diagnosis of refractory anemia with excess of blasts (RAEB) was made. Subsequently, in November 1996, she developed acute myelogenous leukemia (AML), M4 type by FAB classification. The karyotype of MDS and AML clones involved inversion (3) (q21q26) and monosomy 7. The EVI 1 gene was examined and was proved to be rearranged and activated. This may be the first case among the therapy-related cases of MDS/AML reported whose karyotypes were followed and in which the mRNA expression of EVI 1 gene involved was directly proved in the leukemogenesis process of chemotherapy-induced secondary MDS and AML. PMID- 9861237 TI - Pure red cell aplasia and myasthenia gravis with thymoma: a case report and review of the literature. AB - A case of pure red cell aplasia (PRCA), myasthenia gravis (MG) and thymoma is reported. A 70-year-old woman presented with severe anemia. She had been diagnosed as having MG 8 years earlier and her symptoms were adequately controlled with ambenonium chloride. When she visited our hospital, her hematocrit was 13.7% with a hemoglobin concentration of 4.7 g/dl and her reticulocyte counts were persistently abnormal at 0.1%. Although both direct and indirect Coombs' tests were positive, there was no evidence of hemolysis. Routine screening tests for other etiologies of anemia were negative. Serological tests for anti-DNA and anti-acetylcholine receptor antibodies gave positive results. A bone marrow examination revealed severe erythroid hypoplasia. PRCA was diagnosed and the patient was treated with periodic transfusions. A lateral view chest roentgenogram and a computed tomography scan of the thorax showed the presence of an anterior mediastinal mass which was suspected to be thymoma. The patient underwent thymothymectomy and the tumor was diagnosed as a thymoma. Although the patient received no treatment for MG and PRCA after surgery, her hematological test results rapidly improved and she was discharged from the hospital on the 29th postoperative day. At that time, her hematocrit was 33.2%, her hemoglobin concentration was 10.0 g/dl, her peripheral reticulocyte level was 1.8% and her left partial ptosis had improved. She is doing well, 9 months after surgery. For a patient to remain in remission without treatment for PRCA and MG after thymothymectomy is extremely rare. PMID- 9861238 TI - Nasopharyngeal cancer with neck recurrence at 8 years and a lung metastasis at 15 years after the first definitive radiotherapy: a case report. AB - We report a patient with nasopharyngeal cancer with long-term follow-up of more than 16 years after the first course of radiotherapy in 1981. He developed a lung metastasis in 1996 after having a second course of radiotherapy for neck recurrence in 1989. The patient was a 42-year-old man with a nasopharyngeal tumor and a fixed upper neck metastasis (T1N1M0), which was treated with definitive radiotherapy. He manifested regional recurrence, at the margin of the radiation portal, with an 8 year disease-free interval, which was treated successfully by definitive re-irradiation. He developed a solitary lung metastasis, which was treated by video-assisted thoracoscopic lung resection, 7 years disease-free after the second course of radiotherapy. For 20 months after the removal of the lung metastasis he has been generally well without any signs of recurrence of sequelae. This case indicates the efficacy of definitive re-irradiation for regional recurrence and the necessity for long-term observation after radiation therapy for nasopharyngeal cancer. PMID- 9861239 TI - Phyllodes tumor showing intracystic growth: a case report. AB - A phyllodes tumor often grows rapidly and occasionally contains microcysts; however, the tumor rarely shows a morphologically intracystic pattern. We experienced a rare case of a phyllodes tumor with a solid mass growing into the cyst. A 62-year-old female noticed a tumor in her right breast in January 1995. The tumor grew rapidly and she visited our out-patient clinic in February 1995. On physical examination, a 10 x 8 cm, well defined and movable mass with a smooth surface was palpated in the upper outer quadrant of the right breast. Mammography showed a large tumor shadow in the upper outer quadrant of the right breast without any microcalcification. Ultrasonography revealed a large cystic shadow with a low echoic lesion and solid component with heterogeneous internal echo in the cyst. Under general anesthesia, the tumor was widely excised. The resected specimen was 11.5 x 11 x 11 cm in size and the tumor was not invasive to surrounding tissues. Old bloody fluid was contained within the cyst. The gross appearance showed papillary process protrusions into a central cystic cavity. Histological examination revealed a borderline case of phyllodes tumor. Two years after the operation, she is doing well without any recurrence. PMID- 9861240 TI - Bradycardia induced by irinotecan: a case report. AB - Irinotecan chloride (CPT-11) is a new semi-synthetic camptothecin analogue which has encouraging antitumor activity against various malignancies. The major and unique toxicity of CPT-11 is diarrhea. Cardiovascular toxicity is rare and has not been found in clinical trials performed in Japan except for a very few cases of insignificant tachycardiac arrhythmia. We report a case of a 69-year-old man with recurrent colon cancer who suffered from bradycardia induced by infusion of CPT-11. Other toxicities including hematological toxicity and diarrhea were mild. Pharmacokinetic analysis using a limited sampling model revealed that the occurrence of bradycardia did not correlate with the excess of drug exposure. Although all of the cholinergic actions reported in the literature were mild, cardiotoxicity may come to be a clinically significant problem. If the events were examined more thoroughly, the cholinergic effect may be discovered more frequently. To administer CPT-11 safely needs meticulous monitoring not only for hematological toxicity and diarrhea but also for other cholinergic actions including bradycardia. PMID- 9861241 TI - Centenarians in Japan. PMID- 9861242 TI - Liver cancer mortality in Japan. PMID- 9861243 TI - Physician contracts. Business and legal challenges. AB - There is no doubt that PPMCs are here to stay, at least for the near future. This article in no way addresses all of the legal and clinical concerns that the CNE may have when facing an arrangement with a PPMC. Even so, if the CNE can help forge agreements with PPMCs that are legally reliable, that are fiscally sound, that can be managed efficiently, and that are supportive of quality patient care, he or she will have made an invaluable contribution to the future of healthcare delivery. PMID- 9861244 TI - Colorado Consortium for Nursing Work Force Development. Moving from chaos to collaboration. PMID- 9861245 TI - Subtle and not-so-subtle insensitivity to ethnic diversity. PMID- 9861246 TI - Quality and standards for accreditation of continuing nursing education. PMID- 9861247 TI - Improving the ability to detect the impact of labor on patient outcomes. AB - A key part of resource allocation decisions is determining the impact of staffing on clinical outcomes. A national study involving eight hospitals and 77 units lends support for a previously identified labor assignment phenomena that may contribute to an underestimation of the impact of nursing labor on outcomes if only traditional labor measurement methods are used. The authors suggest actions for nurse administrators and researchers who must address frequent queries as to the adequacy of nursing staffing. PMID- 9861248 TI - The influence of education and personality on risk propensity in nurse managers. AB - The current healthcare environment is demanding innovation. Nurse managers are positioned strategically to creatively change and innovate the patient care delivery process, if they are willing to take risks. The authors discuss a study that tests whether a nurse manager's education, years of experience, and personality are related to their propensity to take risks in administrative decision making. Results of the study indicate that nurse managers who achieved at least a bachelors degree, had higher autonomy orientations, and had lower control orientations were more likely to choose higher risk options in nursing administrative decisions. The authors found that self-esteem and years of experience were not related to nurse manager risk propensity. PMID- 9861249 TI - Analysis of performance measures. A veterans home case study. AB - The development or adaptation of a comprehensive performance measurement system is vital for the proper assessment of organizational performance. The authors describe their experience in developing and implementing a modified performance measurement questionnaire for the assessment of an existing performance measurement system in a state veterans home. Adapted for a long-term care environment, the questionnaire determines whether the organization's strategies, performance measurement system, and employees' actions match up and support each other. PMID- 9861250 TI - Understanding Generation X employees. AB - Understanding Generation X employees--those born between 1961 and 1981--is essential if they are to be recruited into and retained in nursing and their potential maximized. The author discusses the times, characteristics, and work values and demands of Generation X. Armed with an enhanced understanding, nurse administrators are better prepared to maximize the potential of Generation X employees. PMID- 9861251 TI - Problem-solving tools for analyzing system problems. The affinity map and the relationship diagram. AB - The author describes how to use two management tools, an affinity map and a relationship diagram, to define and analyze aspects of a complex problem in a system. The affinity map identifies the key influencing elements of the problem, whereas the relationship diagram helps to identify the area that is the most important element of the issue. Managers can use the tools to draw a map of problem drivers, graphically display the drivers in a diagram, and use the diagram to develop a cause-and-effect relationship. PMID- 9861252 TI - Tobacco legislation: time to right some wrong. PMID- 9861253 TI - Remember your office at gift-giving time. It won't hurt if selections are tax deductible. PMID- 9861254 TI - Interferon therapy for hepatitis B and C. PMID- 9861255 TI - Another toxic shock syndrome. Streptococcal infection is even more dangerous than the staphylococcal form. AB - In the battery of diseases caused by group A streptococci, streptococcal toxic shock syndrome is among the most severe. Its nonspecific presentation and rapid progression require that the treating physician be both knowledgeable and vigilant. Certain signs and symptoms suggest that caution is in order: Pain out of proportion to physical findings may herald deep tissue involvement in a patient who at first glance appears to have cellulitis. Streptococcal infection accompanied by hypotension, confusion, or unexplained acute renal insufficiency is a clue to streptococcal toxic shock syndrome. Treatment consists of antibiotic and supportive care, with aggressive surgical debridement of soft-tissue foci of infection when necessary. Anecdotal evidence suggests that intravenous immunoglobulin may have a place in neutralizing the secreted streptococcal toxins that are thought to mediate features of the disease. At present, even with aggressive therapy, the mortality rate of streptococcal toxic shock syndrome can exceed 50%. Understanding of its pathogenesis is progressing. However, until effective interventions are developed, early detection appears to be the best weapon. PMID- 9861256 TI - Trends in smoking-related diseases. Why smoking cessation is still the best medicine. AB - Although millions of Americans have kicked the habit, the effects of cigarette smoking likely will be around for a long time. What was once regarded as a glamorous habit is now recognized as a health threat and an economic burden. But what headway has been made in the reduction of related morbidity and mortality? The authors of this article review the current epidemiologic data on smoking related diseases and make an indisputable case for smoking cessation. PMID- 9861258 TI - Counseling patients to quit smoking. What to say, when to say it, and how to use your time to advantage. AB - Helping patients stop smoking is one of the most important--and frustrating- services primary care physicians can provide. Many physicians are reluctant to spend time talking about tobacco use with patients who show little or no interest in changing their habits. Fortunately, specific technique have been identified that can make the task easier. This article from the distinguished Nicotine Dependence Center of the Mayo Clinic looks at the "best practices" for dealing with nicotine dependence. PMID- 9861257 TI - Drug therapy to aid in smoking cessation. Tips on maximizing patients' chances for success. AB - The arsenal of pharmacologic agents available for smoking cessation has expanded in the last few years, and it is likely to continue to do so. It is important that practicing physicians keep abreast of new methods as they become available and encourage patients who smoke to undertake cessation measures. Nicotine replacement therapy is available in gum, patch, nasal spray, or inhaler form, and bupropion therapy aids in smoking cessation through dopaminergic activity. The foundation of effective intervention is likely to remain unchanged: an individualized plan addressing behavioral, addictive, pharmacologic, and relapse prevention components. In addition to the necessary information about treatment choices, physicians should offer motivation, support, and follow-up to their patients who wish to quit smoking. PMID- 9861259 TI - Assessment of valvular heart disease. Why echocardiography is an essential component. AB - Assessment of patients with valvular heart disease has undergone many changes in the past few decades with the introduction of noninvasive techniques such as echocardiography. Along with the history, physical examination, electrocardiogram, and chest films, echocardiography is an important component of the assessment, providing essential information for diagnosis and follow-up. PMID- 9861260 TI - A puzzling case of palpable hyperpigmentation. Polythelia, or supernumerary nipple. PMID- 9861261 TI - Stopping progression to tophaceous gout. When and how to use urate-lowering therapy. AB - Although the prevalence of tophaceous gout has decreased in the past few years, the disease still exists, and without accurate diagnosis and therapy, it can still result in destructive arthritis. However, use of urate-lowering drugs may reduce plasma urate concentrations enough to allow resorption of tophi and prevent painful tophaceous gout. Some patients may have mechanical problems from tophi, despite adequate control of acute arthritis; in such situations, joint replacement or surgical excision of tophi may be necessary. PMID- 9861262 TI - 10 years of Mectizan. PMID- 9861263 TI - Onchocercal eye disease and the impact of Mectizan treatment. AB - Onchocerciasis continues to be a major cause of blindness, particularly in those sub-Saharan African countries which are outside the area of West Africa monitored by the Onchocerciasis Control Programme (OCP). Onchocercal ocular disease and blindness develop as a result of long exposure to onchocercal infection. Until 1987, suramin and diethylcarbamazine were the only drugs available for the treatment of onchocerciasis and they could not be used for community therapy because of their toxicity and the dosage schedules required. The registration of Mectizan (ivermectin, MSD) for treatment of human onchocerciasis in 1987, and the donation of this drug by Merck & Co. for as long as it is needed, provided a new opportunity for the safe treatment and control of the disease. The data available on the impact of repeated doses of Mectizan on ocular onchocercal disease indicate a significant reduction of ocular microfilarial loads and regression of early lesions of the anterior segment, including iridocyclitis and sclerosing keratitis. Such improvements are seen more rapidly when Mectizan is used than when onchocerciasis is limited by vector control alone. Mectizan treatment also has a beneficial effect on onchocercal optic-nerve disease and visual-field loss. Long-term maintenance of Mectizan therapy should lead to a reduction in the prevalence of blindness in endemic communities. PMID- 9861264 TI - Delivery systems and cost recovery in Mectizan treatment for onchocerciasis. AB - The efficiency of on-going delivery systems and cost recovery in Mectizan (ivermectin, MSD) treatment for onchocerciasis are reviewed. The search is on for an effective system of Mectizan delivery, involving drug procurement, delivery from port to districts and distribution to eligible persons, which can be sustained by the endemic countries for many years. The mechanisms for procuring and clearing the drug at the ports, and the drug's integration into the existing delivery systems of each national health service, need to be improved. Although large-scale treatments by mobile teams or community-based methods evidently achieve high and satisfactory rates of coverage, they also incur high recurrent costs which have to be covered by external partners and are not sustainable by national health services. Cost-sharing is considered an important factor in a sustainable delivery system and community-directed treatment, in which the community shares the cost and ownership of local distribution and is empowered to design and implement it, is likely to be more cost-effective and sustainable. PMID- 9861265 TI - Economic impact of onchocerciasis control through the African Programme for Onchocerciasis Control: an overview. AB - This note overviews several studies that have been conducted on the economic impact of onchocerciasis (river blindness) control through the African Programme for Onchocerciasis Control (APOC). A cost-benefit analysis of the APOC concludes that the programme is highly cost-effective. The economic rate of return (ERR) is 17% if benefits are considered in accordance with the stated objective of the programme (i.e. the achievement of long-term, sustainable, ivermectin-delivery systems). However, the cost-benefit analysis significantly under-estimates the net benefits from the APOC, since it considers, for ease of quantification, only the reduction in blindness as the principal benefit accruing from control activities. Recent studies, summarized here, have shown that there may be substantial benefits (in terms of enhanced productivity, increased household level welfare, and reduced health-related expenditure, for instance) resulting from the reduction of the skin-related symptoms associated with the disease. PMID- 9861266 TI - Diagnostics in onchocerciasis: future challenges. AB - The classical method of determining the prevalence and intensity of onchocercal infection is by the demonstration and counting of microfilariae in biopsies obtained by skin snipping. Although very specific, this technique is inadequate for detecting early, light or prepatent infections, and is also becoming increasingly unacceptable to the populations investigated. The prolonged clearing effect that Mectizan (ivermectin, MSD) treatment has on skin microfilariae also renders the skin-snip method of diagnosis less appropriate in areas with Mectizan treatment. Given all these factors, the greater challenge in the area of diagnostics for onchocerciasis is to develop a less invasive, adequately sensitive, and equally specific diagnostic test, either to replace or to be an adjunct to the present skin-snip method. This challenge is being addressed, with at least three new diagnostic tests for onchocerciasis under development: an immunological assay, based on a three-antigen cocktail; a PCR-based assay, which may also be used for 'pool screening' of blackflies; and the diethylcarbamazine (DEC) patch test. Of all these tests, the DEC patch test seems to fit best the criteria of an ideal test. The PCR assay would be better than the patch test if the cost of using it could be reduced substantially. PMID- 9861267 TI - The impact of Mectizan on the transmission of onchocerciasis. AB - For many years there was no suitable drug available for the control of onchocerciasis. The advent of Mectizan (ivermectin, MSD; an effective microfilaricide), its registration in October 1987 for the treatment of human onchocerciasis, and its suitability for large-scale application were major break throughs in the control of human onchocerciasis via chemotherapy. Several studies, both fly-feeding experiments and community trials, have established that Mectizan treatment causes a significant reduction in the transmission of infection. Although long-term treatment in some isolated foci (such as occur in the New World and in some hypo- and meso-endemic areas elsewhere) appears to interrupt transmission, more prolonged treatment is required to prove if transmission can be stopped. Advantage could be taken of the significant impact of Mectizan on transmission by giving treatment while or just before transmission by blackflies is most intense. PMID- 9861268 TI - Changes in the use profile of Mectizan: 1987-1997. AB - The usually conservative approach of Merck & Co. to drug development became even more so in the Mectizan (ivermectin, MSD) programme because of adverse experiences following 'extra-label' use in Collie dogs and the discovery of a low threshold for acute neurotoxicity in CF-1 mice. Although a very cautious approach and rapid development programme ensued, Merck remained conservative and excluded children under the age of 5 years, pregnant women, and mother who were nursing children under the age of 3 months from treatment. A subsequent, more relaxed set of standards was based on vast human clinical experience, inadvertent use in hundreds of pregnant women without ill-effect, and new laboratory information indicating that the presence of a protective blood-brain barrier protein component (P-glycoprotein) helped to stop Mectizan from crossing the placenta and from crossing the blood-brain barrier in most animal species, including humans. This has allowed more groups to be included in Mectizan treatments: pregnant women living in areas where the risk of loss of sight because of onchocerciasis is very high; and women who are nursing children as young as 1 week of age. Mass distribution of the drug continues to be largely under community control and the likelihood of serious adverse experiences related to finding a human population with unusually low levels of P-glycoprotein (or no P-glycoprotein) seems remote. PMID- 9861269 TI - Control of onchocerciasis: challenges for the future. AB - The control of onchocerciasis has grown over the last two decades to attain global dimensions. This rapid growth poses challenges that are technical and managerial in nature. Appropriate control measures have to be applied to eliminate the disease as a worldwide, public-health problem and prevent the problem recurring. Novel tools and innovative approaches, both for control and surveillance activities, will have to be developed. The health systems of the affected countries need not only to be directly involved in control activities but also to direct operations and have adequate resources to run them successfully. PMID- 9861270 TI - The Mectizan Donation Programme--a 10-year report. AB - In the 10 years since 1987 when Merck & Co. announced a plan to donate its safe and effective microfilaricide, Mectizan (ivermectin, MSD), to treat onchocerciasis wherever and for as long as needed, more than 96 million treatments have been enabled in community-based treatment programmes in all 34 countries, in Africa, the Middle East, and Latin America, where the disease is endemic. It is expected that donations enabling some 33 million treatments will be approved in 1997. Since the beginning of the donation programme, it is estimated that some 19-20 million people have received at least one dose of the drug and that many have received their sixth, seventh, eighth, or even ninth annual dose. PMID- 9861271 TI - Mectizan as a stimulus for development of novel partnerships: the international organization's perspective. AB - Following the free donation of Mectizan (ivermectin, MSD) by Merck & Co. in 1987, early efforts to mass distribute the drug came from non-governmental development organizations (NGDO), which had already established projects in Africa and Latin America by 1989. In the beginning, these projects were NGDO-specific and fairly independent from one another. Subsequently, the need to co-ordinate all attempts at Mectizan distribution led to the creation of the Onchocerciasis Eradication Programme of the Americas (OEPA), in 1991, and the NGDO Co-ordination Group for Ivermectin Distribution, in 1992. The latter was to become the 'prime mover' in Mectizan-distribution programmes, particularly in areas of Africa not monitored by the Onchocerciasis Control Programme in West Africa (OCP), until the advent of the African Programme for Onchocerciasis Control (APOC) in 1995. Once the co ordination group had been established and its co-ordinator appointed, the Mectizan-distribution programme expanded rapidly, reaching 7 million people by 1995. However, the limited resources of the co-ordination group and the need to achieve a better co-ordinated and more comprehensive control of onchocerciasis ultimately led to the creation of the APOC in 1995. The international co operation behind the APOC has inaugurated a unique type of global partnership, in which the success of the programme lies not with a single player, but with close working relationships and collaboration between many partners: donors; governments of endemic countries; NGDO; international organizations; and private institutions. This is a complex process, the success of which cannot be guaranteed. However, if it can be made to succeed, it will serve as a model to be used to address other serious and intractable development or health problems. PMID- 9861272 TI - Human onchocerciasis: current assessment of the disease burden in Nigeria by rapid epidemiological mapping. AB - Rapid epidemiological mapping of onchocerciasis (REMO) was carried out in Nigeria between 1994 and 1996, to provide accurate estimates of the populations living in high-risk areas for the disease and to identify the communities to be given priority treatment with Mectizan (ivermectin, MSD). The rapid epidemiological assessment (REA) teams which carried out the survey comprised epidemiologists, geographers, and entomologists. They used topographical maps, at a scale of 1:250,000, to select sample villages at 30-km intervals along selected river courses. Several non-governmental development organizations participated in the surveys, providing support to the teams. In each study community, 50 adult males who had lived in the area for at least 10 years were randomly selected and examined for the presence of palpable, onchocercal nodules. The REA results were cross-validated by an independent REMO expert and then presented, initially as percentage prevalences in pie-chart form, using a geographical information system. The number of Nigerians living in high-risk areas (demarcated by river systems with villages that had > or = 19% prevalence) and who therefore require urgent Mectizan treatment, was estimated as 13,809,313. The national REMO map produced is allowing the African Programme for Onchocerciasis Control's technical consultative committee to prioritize the approval of proposals submitted from Nigeria in a rational and effective manner. PMID- 9861273 TI - Onchocercal dermatitis: clinical impact. AB - The clinical features of onchocercal dermatitis include itching, papular and papulomacular rash, skin atrophy and depigmentation. The results of a multi country study have shown that > 30% of the population in communities where onchocerciasis is endemic have onchocercal dermatitis. The most troubling symptom suffered by those affected was itching and this was closely related to reactive onchocercal dermatitis (acute papular, chronic papular and/or lichenified lesions). Reactive onchocercal dermatitis and troublesome itching were common in all age-groups and were an important cause of stigma in most endemic communities, those affected suffering from poor self-esteem. Concern about onchocercal depigmentation varied between study sites and subjects. PMID- 9861274 TI - Mectizan delivery systems and cost recovery in the Central African Republic. AB - The Central African Republic (CAR) has a serious onchocerciasis problem. The disease is endemic in three quarters of the country and there is considerable onchocercal blindness in the north-west. The low population density and extreme poverty (the CAR being one of the 20 poorest countries in the world) combine to make mass treatment with Mectizan (ivermectin, MSD) a challenge. Although planned, primary health care (PHC) is not widely developed in the country. Mectizan distribution was carried out in 1993-1994 by mobile teams in order to address the most urgent need, particularly in the north-west. Since then, the strategy has been one of community involvement, using village health workers, chosen by their own communities, to do the treatment. The system has been a stimulus to the development of PHC in some areas, as the co-ordinators of the Mectizan programme are often the only health personnel to visit every village. The long distances between health centres, with a mean of 45 km, are likely to be an obstacle to the population collecting their own Mectizan, within the self treatment system otherwise in place. Operational research is planned to examine ways in which the population can contribute to cost recovery without there being a reduction in treatment coverage. PMID- 9861275 TI - Onchocerciasis: the burden of disease. AB - The greatest burdens related to human onchocerciasis are the result of the eye and skin lesions and severe itching produced by the microfilariae. Although the major manifestations of the disease do show geographical variation (e.g. onchocercal blindness is not a common complication in all endemic countries), they are often sufficiently severe to prevent human use of the often very fertile land close to the rivers in which the vectors breed. Though for many years thought to be of relatively minor importance compared with onchocercal eye disease, the skin lesions of onchocerciasis have recently been shown to be a major socio-economic burden, in terms of disability-adjusted life-years. The demonstration of an excellent correlation between the prevalence of palpable nodules in a community and the community microfilarial load has led to the development of rapid, safe and non-invasive methods to assess and map the levels of endemicity across whole countries. This has enabled mass treatment with Mectizan (ivermectin, MSD) to be targeted first at hyperendemic communities. Estimates of the burden of onchocerciasis will continue to change as better means of measurement become available. It seems possible, however, that use of Mectizan will eliminate the disease before its true burden can be estimated accurately. PMID- 9861276 TI - Onchocerciasis: changes in transmission in Mexico. AB - There are now three endemic foci of onchocerciasis in Mexico, all located in mountainous areas in the south-east: two in the state of Chiapas and one in the state of Oaxaca. Together, these three foci cover 16,900 km2 and contain about 286,000 people in 947 localities, most of the localities being small and scattered. The main economic activity in all the foci is coffee farming. Introduction of Mectizan (ivermectin, MSD) into Mexico in 1989 revolutionized control of onchocerciasis in the country and made elimination of the disease a reasonable goal. Concerted efforts between 1989 and 1997, supported by Merck & Co., the River Blindness Foundation and the Onchocerciasis Elimination Programme in the Americas, have led to steady decreases in the incidence of new cases, nodule prevalence and skin-snip positivities. The improvement has been most marked in the Oaxaca focus, where transmission may have been interrupted. As prior to attempts at control, the largest populations at risk are in the Chiapas foci, where logistical problems and migration have combined to limit the success of local control programmes. PMID- 9861277 TI - Problems and perspectives of managing an onchocerciasis control programme: a case study from Plateau state, Nigeria. AB - The onchocerciasis control programme in Plateau state (now Plateau and Nasarawa states), Nigeria, was one of the pioneering Mectizan-distribution projects in Nigeria. Although initiated under the River Blindness Foundation (RBF) in 1991, in collaboration with the Ministry of Health, it was absorbed into the Carter Center's Global 2000 River Blindness Programme (GRBP) in 1996. The objectives of the programme were to support the delivery of Mectizan (ivermectin, MSD) to at least 80% of those living in communities where onchocerciasis was highly endemic, within the first 3 years of the project's inception, and to maintain this coverage for a period of 10-15 years. The programme has so far been successful, and much of this success is attributed to problem identification and problem solving through continuous review and evaluation of programme activities, and implementation of strategies, when required, to ensure those programme objectives are met. The implementation steps of the programme, and some of the managerial problems identified during the course of the effort, are reviewed. The challenge now is to learn how to transform this functional, programme-designed and programme-directed effort into the new community-directed treatment being promoted by the African Programme for Onchocerciasis Control. The new challenges of the transition require middle-level managers and implementors with effective, efficient and indeed state-of-the-art management skills. PMID- 9861278 TI - The burden of Onchocerca volvulus in Sudan. AB - Onchocerciasis has been reported in Sudan since 1908 and now prevails in three endemic regions known as the southern, northern and eastern foci. The southern focus is the largest, with nodule and blindness rates exceeding 80% and 12%, respectively, in certain villages. Onchocercal infection in this region causes only a mild skin reaction although microfilarial loads in the skin are high. In contrast, those with onchocerciasis in the northern focus, located between the fourth and fifth Nile cataracts, present with limited but severe skin reactions, low nodule rates (16%), low microfilarial loads in the skin and no ocular involvement. The characteristics of patients from the eastern focus, close to the border with Ethiopian border are similar to those in the north, although most onchocercal skin disease in this area comprises the severe localized pruritus known as sowda. PMID- 9861279 TI - The burden of onchocerciasis in Uganda. AB - Onchocerciasis is far more prevalent and far more of a public-health and socio economic problem in Uganda than it was thought to be a decade ago. It appears that over a million Ugandans have the disease and nearly two million others live in endemic areas. A few years of annual treatment with Mectizan (ivermectin, MSD) have reduced the community microfilarial load (CMFL) in sentinel villages in Uganda by 52%-100%. The greatest reductions (100%) have been in those villages with the lower CMFL pre-treatment. Assessments at intervals of much less than 1 year should help to show how quickly skin cleared by Mectizan is re-invaded by microfilariae and give an idea of how frequently treatment should be given to produce the greatest benefits. It seems likely that it would be best to give Mectizan at intervals of < 1 year and that such frequent treatment only becomes feasible if Mectizan distribution is community-directed. PMID- 9861280 TI - A multi-centre study of the effect of Mectizan treatment on onchocercal skin disease: clinical findings. AB - A multi-centre, double-blind, placebo-controlled trial of the effect of Mectizan (ivermectin, MSD) treatment on the skin disease and severe itching associated with onchocerciasis was carried out in the forest zones of Nigeria, Ghana and Uganda. Overall, 4072 subjects, none of whom had received Mectizan previously, were enrolled and allocated into four groups, to receive Mectizan every 3, 6 or 12 months or placebo every 3 months. Subjects with skin lesions were stratified within each treatment group to ensure equal representation. Each subject was given a clinical examination and interviewed on enrolment and then 3-monthly for 15 months. The presence and severity of itching were determined by open-ended questions followed by probing questions. Skin lesions were classified and their severity graded using a standard system. Analysis of the results was restricted to the data from the 1530 subjects who received all their scheduled treatments and attended all the follow-up visits. From 6 months onwards, all subjects who had received Mectizan reported less severe itching, had lower prevalences of reactive skin lesions and had less severe skin lesions than those in the placebo group (P < 0.05 for each). The greatest reductions, in both itching and skin disease, were seen in subjects treated with Mectizan every 6 months. Mectizan treatment is therefore beneficial for the control of troublesome itching and for reducing the prevalence and severity of skin disease causes by Onchocerca. it is recommended that mass distribution of Mectizan to communities in the study areas be carried out at 6-monthly intervals for maximum efficacy. PMID- 9861281 TI - Challenges for the future: loiasis. PMID- 9861282 TI - Corporate donations. PMID- 9861283 TI - Partnerships between non-governmental development organizations. PMID- 9861284 TI - Onchocerciasis: a Latin American perspective. PMID- 9861285 TI - The donation of Mectizan. PMID- 9861286 TI - Cameroon and Chad: cost recovery. PMID- 9861287 TI - Combating onchocerciasis in Africa after 2002: the place of vector control. PMID- 9861288 TI - Problems and perspective in programme management: the case of the National Onchocerciasis Control Programme in Nigeria. PMID- 9861289 TI - Community perspective on Mectizan's role as a catalyst for the formation of novel partnerships. PMID- 9861290 TI - The challenge of establishing community-directed treatment with Mectizan in Uganda. PMID- 9861291 TI - Animal models--Onchocerca ochengi and the development of chemotherapeutic and chemoprophylactic agents for onchocerciasis. PMID- 9861292 TI - Endothelins. PMID- 9861293 TI - The discovery of endothelins. PMID- 9861294 TI - Endothelin in hypertension: a role for receptor antagonists? AB - The rapid development of endothelin-receptor antagonists has made the endothelin pathway a new therapeutic target in the treatment of cardiovascular diseases, only ten years after the report of its discovery. While the first clinical trials will help to position this new family of compounds in our therapeutic armament for the treatment of essential or secondary forms of hypertension, several preclinical chronic studies already provide a picture of what we can expect from these drugs. Endothelin-receptor antagonists are not effective in all experimental models of hypertension, but those that respond present hypertrophy of small arteries, secondary to a local overexpression of the peptide. Although angiotensin II seems to represent a stimulus for endothelin overexpression in some models, other, as yet undetermined, stimuli are likely in others. Besides their narrow spectrum of antihypertensive activity, endothelin-receptor antagonists may also protect from complications of hypertension by improving end organ function in a pressure-independent manner. This seems to be the case for the structure and reactivity of resistance arteries, as well as for renal damage. However, it is not clear at this point if cardiac structure and function are improved beyond the benefits produced by blood pressure reduction. The first results in essential hypertensive subjects suggest some degree of efficacy of endothelin-receptor antagonists. Other clinical trials will help to determine if secondary forms of the disease benefit equally or more from this new class of drugs, and if end-organ damage can be reduced beyond blood-pressure reduction. PMID- 9861295 TI - Endothelin blockers and renal protection: a new strategy to prevent end-organ damage in cardiovascular disease? PMID- 9861297 TI - Endothelin and heart transplantation. PMID- 9861296 TI - Endothelin and restenosis. PMID- 9861298 TI - Endothelin-1 has haemodynamic effects at pathophysiological concentrations in patients with left ventricular dysfunction. AB - OBJECTIVES: Plasma levels of immunoreactive endothelin-1 (ET-1) are raised in chronic heart failure. Whether plasma ET-1 contributes to the haemodynamic derangement found in chronic heart failure is not known. We investigated the effects of exogenous ET-1 on the pulmonary and systemic vasculature in patients with left ventricular systolic dysfunction (LVD), with or without overt heart failure. METHODS: ET-1 was infused at 1, 5 and 15 pmol/min into a distal pulmonary artery of ten patients with LVD to achieve plasma concentrations of ET 1 similar to those found in patients with heart failure and pulmonary hypertension. Haemodynamics were measured using a pulmonary thermodilution catheter and an arterial line. Intravascular Doppler and local pulmonary angiography were used to assess local pulmonary blood flow in the first four patients. RESULTS: Systemic haemodynamic changes occurred with ET-1 infusion: mean arterial pressure (100 +/- 3 [standard error of the mean]) to 107 +/- 3 mmHg; p < 0.01) and systemic vascular resistance (1699 +/- 118 to 2033 +/- 135 dynes s/cm5; p < 0.001) rose, while the cardiac index fell from 2.43 +/- 0.17 to 2.20 +/- 0.16 l/min/m2 (p < 0.002). Mean pulmonary artery pressure (21 +/- 2 mmHg) and pulmonary vascular resistance (151 +/- 14 to 147 +/- 14 dynes s/cm5) did not change however. CONCLUSIONS: Exogenous ET-1, when infused to achieve plasma concentrations similar to those in severe heart failure and pulmonary hypertension, causes systemic but not pulmonary vasoconstriction. PMID- 9861300 TI - Acute effects of an endothelin-1 receptor antagonist bosentan at different stages of heart failure in conscious dogs. AB - OBJECTIVE: Inhibition by endothelin antagonist is a potential therapy in heart failure. However, the effect of endothelin inhibition during the development of heart failure has not been evaluated. The goal of our study was to examine the acute hemodynamic effects of the mixed endothelin receptor antagonist bosentan in the control state and at different stages of heart failure induced by right ventricular pacing (250 bpm) in conscious dogs. METHODS: Nine dogs were chronically instrumented for the measurements of left ventricular pressure and its first derivative (dP/dt), cardiac output, left ventricular regional wall thickness and aortic pressure. Bosentan (3 mg/kg, i.v. bolus) and placebo were given at control, at 1 week of pacing (stage of left ventricular dysfunction with perserved cardiac output) and at 3 weeks of pacing (phase of heart failure with low cardiac output). RESULTS: With the development of heart failure, baseline plasma endothelin level increased progressively. Placebo did not induce hemodynamic and plasma endothelin changes during the 30 min recording at any stage. At control, bosentan did not change hemodynamics. At 1 and 3 weeks of pacing, bosentan did not modify left ventricular myocardial function indices but reduced mean arterial pressure (by 7 +/- 2 and 8 +/- 1 mm Hg respectively, p < 0.005). Bosentan increased stroke volume at 3 weeks of pacing only. CONCLUSIONS: Endothelin inhibition by endothelin antagonist bosentan, decreases aortic pressure in both early left ventricular dysfunction and in heart failure in contrast with the control state. In the phase of heart failure with low cardiac output, bosentan increases stroke volume. In the early left ventricular dysfunction, bosentan, by reducing arterial pressure, may limit the deterioration of cardiac function through a reduction of the workload imposed on the heart. PMID- 9861299 TI - Beneficial effects of long-term selective endothelin type A receptor blockade in canine experimental heart failure. AB - OBJECTIVES: We examined the effects of chronic type A endothelin receptor (ETA) blockade in a dog model of pacing-induced cardiomyopathy. METHODS: Eight dogs received an ETA antagonist, LU 135252 (50 mg/kg orally daily) and nine dogs received a matching placebo starting at day three of pacing and continued for the remainder of the three weeks of pacing. RESULTS: In the placebo group, the mean pulmonary artery pressure and left ventricular end diastolic pressure increased from 16 +/- 3 and 8 +/- 2 mmHg, respectively, at baseline to 40 +/- 11 and 34 +/- 7 mmHg, respectively, at two weeks (both p < 0.001 versus baseline). Cardiac output declined from 3.5 +/- 0.7 to 1.9 +/- 0.6 l/min (p < 0.001). In the treatment group, LU 135252 attenuated the increase in mean pulmonary artery and left ventricular end diastolic pressure (16 +/- 3 and 9 +/- 1 mmHg at baseline to 29 +/- 3 and 27 +/- 3 mmHg, respectively, at two weeks (p < 0.001), and the decline in cardiac output (3.2 +/- 0.3 to 2.6 +/- 0.8 l/min, p < 0.01; p < 0.05 versus placebo for the three parameters). Systemic and pulmonary vascular resistance increased only in the placebo group. Left ventricular end-diastolic volume increased to a similar degree. However, LU 135252 attenuated the increase in plasma norepinephrine level (placebo, 1.2 +/- 0.5 to 3.7 +/- 1.9 pmol/l; treatment, 0.8 +/- 0.3 to 2.4 +/- 0.6 pmol/l; both p < 0.001 versus baseline; p < 0.05 versus placebo). CONCLUSION: Our results suggest that endothelin-1 plays a role in the hemodynamic perturbations in canine pacing-induced cardiomyopathy. The favourable hemodynamic effects without concomitant aggravation of neurohormonal activation suggests that ETA receptor blockade may be beneficial in the treatment of heart failure. PMID- 9861301 TI - Altered inotropic response of endothelin-1 in cardiomyocytes from rats with isoproterenol-induced cardiomyopathy. AB - OBJECTIVE: The positive inotropic effect of endothelin-1 (ET-1) on normal myocardial contraction may be altered in pathological states. The purpose of this study was to assess the direct effect of ET-1 on cardiomyocyte performance and its cellular mechanism in congestive heart failure (CHF). METHODS: We measured the plasma levels of ET-1 and compared the effects of ET-1 (10(-10)-10(-8) M) on contractile performance and the [Ca2+]i transient in the myocytes of left ventricles (LV) from 15 age-matched normal adult rats and 15 rats with isoproterenol (ISO)-induced CHF. RESULTS: With CHF, the plasma levels of ET-1 (19.7 +/- 6.3 vs. 4.1 +/- 0.5 fmol/ml, p < 0.05) were markedly elevated. In normal myocytes, superfusion of ET-1 caused significant increases in the systolic amplitude (SA, 8-16%) and the peak velocity of shortening (dL/dtmax, 20-35%; p < 0.01) without causing a change in the peak [Ca2+]i transient. In contrast, in myocytes from CHF rats, ET-1 produced significant reductions in SA (9-13%) and in the velocity of relengthening, dR/dtmax (10-14%; p < 0.05). The myocytes' dR/dtmax also decreased by 8-10% (p < 0.05). These changes were associated with a significant decrease in the peak [Ca2+]i transient (20-23%, p < 0.01). These responses to ET-1 were abolished by the incubation of myocytes with an ETA receptor antagonist (BQ123) or a protein kinase C (PKC) inhibitor (H-7 or staurosporine). CONCLUSION: ISO-induced CHF is associated with elevated plasma ET 1 and an altered cardiomyocyte response to ET-1. After CHF, ET-1 produces a direct depression of cardiomyocyte contractile performance that is associated with a significant decrease in the peak [Ca2+]i transient. These effects are likely to be mediated through ETA receptors and involve the PKC pathway. PMID- 9861302 TI - Selective ETA receptor blockade prevents left ventricular remodeling and deterioration of cardiac function in experimental heart failure. AB - BACKGROUND: Left ventricular (LV) dilation, which is a predictor of survival in humans with chronic heart failure (CHF), is limited by a mixed endothelin ETA-ETB antagonist. Whether selective ETA receptor blockade influences LV dilation is unknown. We determined, in a rat model of CHF, the effects of the ETA receptor blocker LU 135,252 on LV remodeling. METHODS AND RESULTS: Rats were subjected to coronary artery ligation and treated for ten weeks with placebo or LU 135,252 (LU), at a dose of 10 or 30 mg kg-1 day-1. Systolic blood pressure and heart rate (plethysmography) were determined in conscious animals before and after four and ten weeks of treatment. At these time points, cardiac output and LV dimensions were measured in anesthetized rats by transthoracic echocardiography. LV hemodynamics were determined in anesthetized rats after ten weeks. Pressor responses to ET-1 (1 nmol/kg, i.v.) and sarafotoxin S6c (0.3 ng/kg, i.v.) were measured, to assess the efficacy of ET receptor antagonism and the lack of blockade of ETB receptor blockade, respectively. The pressor response to ET-1 was significantly reduced by LU (% change in systolic blood pressure: sham: 9 +/- 1; CHF: 5 +/- 1; CHF LU: 0 +/- 3 and -4 +/- 2% for the low and high dose, respectively). LU did not affect the response to sarafotoxin (CHF: -37 +/- 3; CHF LU: -29 +/- 3 and -28 +/- 2% for the low and high dose, respectively). Both doses of LU decreased systolic blood pressure, but only the high dose of LU reduced heart rate. Furthermore, LU restored cardiac output dose-dependently throughout the study. Both doses of LU limited LV dilatation and deterioration of LV fractional shortening to the same extent. After ten weeks, LU normalized LV end diastolic- and central venous pressures, but did not affect LV dP/dtmax or dP/dtmin. LU did not prevent the development of cardiac hypertrophy, but reduced LV collagen density. CONCLUSIONS: In this rat model, the selective ETA receptor blocker LU, at the dose of 30 mg kg-1 day-1, reduced blood pressure and heart rate, limited progressive left ventricular remodeling and improved cardiac hemodynamics and function. These effects of LU might have important clinical relevance in the treatment of heart failure. PMID- 9861303 TI - Reduced pulmonary metabolism of endothelin-1 in canine tachycardia-induced heart failure. AB - OBJECTIVES: Plasma endothelin-1 (ET-1) increases in congestive heart failure (CHF). The pulmonary vascular bed could contribute to this increase through a reduced clearance. We evaluated the effect of tachycardia-induced CHF on pulmonary ET-1 kinetics. To discern between changes due to variations in pulmonary hemodynamics from true alterations of endothelial cell functions, we quantified ET-1 kinetics in isolated rat lungs under variable pressure and flow rate conditions. METHODS AND RESULTS: Indicator-dilution studies were performed in anesthetized dogs (n = 14) before and 3 weeks after rapid ventricular pacing and in isolated lungs from healthy rats (n = 4). In isolated lungs, graded increases in perfusion rate from 5-25 ml/min caused gradual reductions in ET-1 extraction from 60 +/- 1.5% to 17 +/- 4.9% (mean +/- S.D.). The capacity to clear ET-1 from the circulation, as computed from the permeability-surface area product (PS), however did not vary over this range of flows. CHF increased plasma ET-1 (11.2 +/- 11.4 vs. 5.2 +/- 1.6 fmol/ml, p < 0.01), did not affect pulmonary ET-1 extraction (29.4 +/- 12.5% vs. 29.9 +/- 12.9%), but decreased the PS (8.3 +/- 5.4 cm3/s vs. 14.4 +/- 9.9 cm3/s, p = 0.038). Contrary to the invariability of the PS in normal isolated rat lungs, CHF was associated with a positive relationship between the PS and pulmonary plasma flow (r = 0.65, p < 0.01). ET-1 binding studies in lung tissues showed no significant variations in ETA and ETB receptors densities but revealed a threefold decrease in binding affinity (p < 0.01) that may explain the reduced clearance. CONCLUSION: CHF causes a reduction of pulmonary ET-1 clearance that likely contributes to the increased circulating ET 1 levels and reflects pulmonary metabolic dysfunction associated with this condition. PMID- 9861304 TI - Comparison of the acute effects of a selective endothelin ETA and a mixed ETA/ETB receptor antagonist in heart failure. AB - OBJECTIVE: Both the selective endothelin (ET) ETA receptor and mixed ETA/ETB receptor antagonists improve haemodynamics in patients and experimental models with congestive heart failure (CHF) and reduce the mortality in CHF rats. However, it remains unclear which of these antagonists is superior in the treatment of CHF. In addition, there is little information as to whether these ET receptor antagonists contribute to the neuroendocrine regulation and body fluid balance. We therefore investigated the cardiorenal and neurohumoral benefits of selective ETA receptor and mixed ETA/ETB receptor antagonists in CHF. METHODS: We administered acutely either the selective ETA receptor antagonist FR139317 (FR, n = 6, 1 and 10 mg/kg) or the mixed ETA/ETB receptor antagonist TAK-044 (TAK, n = 6, 1 and 3 mg/kg) to conscious dogs with CHF induced by rapid right ventricular pacing for ten days. RESULTS: Both FR and TAK decreased the cardiac pressures and the plasma atrial natriuretic peptide level and increased the cardiac output and urinary sodium excretion. FR increased the urine flow rate in association with an increased glomerular filtration rate and renal plasma flow, while TAK reduced the plasma aldosterone level. Neither antagonist increased the plasma renin activity or norepinephrine levels. CONCLUSIONS: These ETA/ETB antagonist. However, the long-term administration of a mixed ETA/ETB receptor antagonist would improve not only the haemodynamics but also prevent fluid retention by suppressing secretion of aldosterone during the treatment of chronic CHF. PMID- 9861305 TI - Endothelin and ischaemic arrhythmias-antiarrhythmic or arrhythmogenic? AB - OBJECTIVE: The aim of this study was to investigate the influence of endogenously released and exogenously applied endothelin-1 (ET-1) on ischaemia-induced arrhythmias. METHODS: Ischaemia was induced in pentobarbitone-anaesthetised rats by ligation of a coronary artery for 30 min. To determine the role of endogenous ET-1 in ischaemic arrhythmias, either the ETA receptor antagonist BQ123 (50 micrograms/kg/min, i.v.; n = 10) or the ETB receptor antagonist PD161721 (0.1 or 1 mg/kg i.v.; n = 10 per group) was administered before the onset of ischaemia. To assess the influence of exogenous ET-1 on arrhythmias, ET-1 (1.6 nmol/kg i.v.) was administered 5 min before ischaemia in the absence (n = 12) or presence of BQ123 (n = 10) or PD161721 (n = 10). The total number of ventricular ectopic beats (VEB's) were counted and expressed as median (Q1-Q3) and the incidence of ventricular fibrillation (VF) and ventricular tachycardia (VT) in each group was determined. Mean arterial blood pressure (MABP) and heart rate (HR) were measured. RESULTS: In control animals (n = 20), the incidence of VF was 65% and the total VEB count was 2775 (1870-4041). Both BQ123 and the higher dose of PD161721 reduced the VEB count to 654 (348-1489; P < 0.05) and 782 (432-1153; P < 0.05) respectively. Only PD161721 reduced the incidence of VF (to 10%; P < 0.05). Administration of ET-1 reduced VEB's to 1530 (1204-2017); P < 0.05) and the incidence of VF to 17% (P < 0.05). Neither PD161721 nor BQ123 modified this antiarrhythmic effect of ET-1, but rather enhanced the reduction in arrhythmias. Before occlusion, ET-1 caused a transient fall in MABP (from 107 +/- 3 to 63 +/- 3 mmHg; P < 0.05). PD161721, but not BQ123, partially blocked this effect. Upon occlusion, MABP fell in control animals (from 106 +/- 4 to 67 +/- 4 mmHg at 1 min post-occlusion; P < 0.05). This was significantly attenuated by ET-1, although neither of the antagonists were able to block this effect of ET-1. CONCLUSIONS: ET-1 released endogenously during ischaemia is arrhythmogenic whereas exogenous application of ET-1 may, under certain conditions, be antiarrhythmic. PMID- 9861306 TI - Pulmonary and cardiac expression of preproendothelin-1 mRNA are increased in heart failure after myocardial infarction in rats. Localization of preproendothelin-1 mRNA and endothelin peptide. AB - OBJECTIVES: Recent reports indicate that endothelin (ET) plays an important pathophysiological role in congestive heart failure (CHF). However, existing data on local cardiopulmonary ET production are few. No studies have hitherto examined the specific anatomic localization of cardiopulmonary ET synthesis in CHF. Thus, the aims of the present study were to examine whether cardiopulmonary preproET-1 mRNA synthesis is upregulated in CHF and to determine the anatomic localization of preproET-1 mRNA and the mature peptide. METHODS: CHF was induced in rats by occluding the left coronary artery. Only animals with a left ventricular end diastolic pressure above 15 mmHg after one week were included (n = 28). Sham operated animals served as controls (n = 24). Hearts and lungs were examined by mRNA slot blot analyses, in situ hybridization (ISH) and immunohistochemistry (IHC). RESULTS: In CHF-rats, slot blot analyses revealed a 3.5 +/- 1.1-fold and a 6.4 +/- 0.8-fold upregulation of preproET-1 mRNA in the noninfarcted and the infarcted area of the left ventricles, respectively (p < 0.05 for both). ISH revealed that the preproET-1 mRNA was localized predominantly over the granulation tissue in the infarcted region. The ET peptide was predominantly localized to inflammatory cells and remaining cardiomyocytes in the infarcted region as determined by IHC. Lungs from CHF-rats showed a 1.5 +/- 0.1-fold upregulation of preproET-1 mRNA (p = 0.01). The most abundant preproET-1 mRNA and ET-1-like-immunoreactivity (ET-1-ir) was seen over inflammatory cells and over airway epithelial cells. Some ET-1-ir was also located to bronchial and vascular smooth muscle cells. CONCLUSION: Increased cardiopulmonary ET synthesis strongly suggest a pathophysiological role for ET in CHF. PMID- 9861307 TI - Insulin and IGF-I attenuate the coronary vasoconstrictor effects of endothelin-1 but not of sarafotoxin 6c. AB - OBJECTIVE: To examine the hypothesis that insulin and insulin-like growth factor I (IGF-I) attenuate endothelin-induced contraction of porcine coronary epicardial arteries in vitro. BACKGROUND: Endothelin-induced coronary vasoconstriction is mediated by two types of receptors, A (ETA) and B (ETB), resulting in calcium influx. Both insulin and IGF-I attenuate endothelin-induced calcium influx into porcine coronary artery smooth muscle. METHODS: Epicardial arteries harvested from juvenile pigs were contracted with cumulative concentrations of endothelin-1 (ETA- and ETB-receptor agonist; 10(-10)-10(-6) M) or of sarafotoxin-6c (ETB receptor agonist; 10(-11)-10(-7) M). In additional experiments, endothelin-1 or sarafotoxin-6c were added after incubation with 10(-8) M regular insulin or IGF I. These experiments were repeated in vessels without endothelium. Contraction for each vessel was calculated relative to the response to 60 mM KCl. RESULTS: The maximal contractions to endothelin-1 in vessels with and without endothelium were 158 +/- 8 and 200 +/- 21%, respectively (p < 0.05 at 10(-8.5)-10(-6.5) M). Both insulin (at 10(-7)-10(-6) M) and IGF-I (at 10(-6.5)-10(-6) M) attenuated the contraction to endothelin-1 in vessels with intact endothelium, as well as in vessels without endothelium (at 10(-7) and 10(-6) M for insulin and 10(-7.5)-10( 6) M for IGF-I). The maximal contractions to sarafotoxin-6c in vessels with and without endothelium were 54 +/- 13 and 84 +/- 7%, respectively (p < 0.05 at 10( 9), 10(-8.5) and 10(-7) M). Insulin and IGF-I did not affect the response to sarafotoxin-6c in vessels with and without endothelium. CONCLUSION: Insulin and IGF-I attenuated ETA-receptor-mediated coronary contraction through an endothelium-independent mechanism. The IGF axis may serve as an endogenous modulator of endothelin-mediated vasoconstriction. PMID- 9861308 TI - Impaired paracrine effect of endothelin-1 on vascular smooth muscle in streptozotocin-diabetic rats. AB - OBJECTIVE: This study was to examine the effect of streptozotocin-induced diabetes on endothelin-1 and its receptors in the mesenteric artery and in the thoracic aorta. METHODS: Diabetes was induced in SD rats by streptozotocin. Insulin was given subcutaneously. Endothelin-1 levels in the plasma, thoracic aorta and mesenteric artery were measured using radioimmunoassay. The Bmax and Kd values of endothelin-1 receptors in the mesenteric artery and in the thoracic aorta were analyzed using Scatchard plot analysis. Preproendothelin mRNA levels were examined using RT-PCR. RESULTS: Endothelin-1 levels in the mesenteric artery (83.6 +/- 6.9 pg/mg protein) and in the thoracic aorta (73.9 +/- 8.2 pg/mg protein) increased in 2 week diabetic rats compared with both control (51.8 +/- 5.3, 46.3 +/- 5.9 pg/mg protein) and insulin treated rats (65.6 +/- 8.1, 48.1 +/- 4.2 pg/mg protein) but not in 4 week diabetic rats. There was no change in plasma endothelin-1 levels in these diabetic rats. The RT-PCR results indicated that preproendothelin mRNA levels in the mesenteric artery (0.38 +/- 0.02 vs 0.52 +/- 0.05 units) and in the thoracic aorta (0.45 +/- 0.06 vs 0.62 +/- 0.03 units) decreased in 4 week diabetic rats but not in 2 week diabetic rats. A significant increase in Kd and Bmax of endothelin receptors in the mesenteric artery and in the thoracic aorta was observed in both 2 week (about 70%) and 4 week (80-85%) diabetic rats. Insulin replacement reversed the effects of diabetes on endothelin 1 peptide contents, preproendothelin mRNA levels and the binding activity in the blood vessels. CONCLUSION: Increased endothelin peptide content with no change in mRNA or decreased mRNA levels with no change in peptide content together with increased receptor binding sites and affinities might imply a decrease in endothelin release and therefore an impaired paracrine effect of endothelin on vascular smooth muscles in these STZ-diabetic rats. PMID- 9861309 TI - Chronic blockade of endothelin ETA receptors improves flow dependent dilation in resistance arteries of hypertensive rats. AB - OBJECTIVE: Flow (shear stress)-induced dilation (FD) is attenuated in hypertension. Flow triggers the release by endothelial cells of dilators, such as NO or cyclo-oxygenase (COX) derivatives and constrictor factors such as endothelin-1 (ET-1) which might be involved in several cardiovascular diseases. We hypothesized that ET-1 might play a functional role in FD and participate in the endothelial dysfunction in hypertension. METHODS: We investigated the effect of chronic treatment with the ETA receptor blocker LU135252 (50 mg/kg/day) for 2 weeks on the dilator response to flow in normotensive (Wistar-Kyoto; WKY) or hypertensive (SHR, n = 7 or 8 per group) rats. RESULTS: Systolic arterial pressure was not significantly affected by chronic ETA receptor blockade in both strains. In mesenteric resistance arteries (diameter: approximately 100 microns), isolated in vitro, FD was lower and myogenic tone higher in SHR than in WKY rats. Chronic ETA receptor blockade increased FD by 73% (7.5 +/- 1.5 to 13.0 +/- 2.7 microns dilation with a flow-rate of 150 microliters/min) in SHR (no effect in WKY). The participation of NO to FD was increased in SHR and the participation of dilator COX product(s) (blocked by indomethacin 10 mumol/l) to FD was significantly increased in SHR and in WKY. In control rats FD was improved by acute ETA receptor blockade in WKY rats (18.5 +/- 2.0 to 23.2 +/- 1.8 microns dilation to flow-rate of 150 microliters/min) and significantly more in SHR (6.0 +/- 1.8 to 15.1 +/- 1.6 microns). Acetylcholine-induced dilation was also improved by chronic ETA receptor blockade (no effect of an acute blockade). Myogenic and phenylephrine-induced tone were not affected by chronic or acute ETA receptor blockade. The improvement of endothelium-dependent dilation was not related to a change in blood pressure. CONCLUSION: Chronic ETA receptor blockade increased flow-induced dilation in SHR possibly by suppressing flow-induced ETA stimulation and by improving the release of dilator products by the endothelium. PMID- 9861310 TI - Endothelin B receptor-mediated vasoconstriction induced by endothelin A receptor antagonist. AB - OBJECTIVE: The vasoconstrictor effect of endothelins (ET) is mediated by endothelin A (ETA) and endothelin B (ETB) receptors. Furthermore, ETB receptor stimulation results in release of vasodilators. Hence, ETA receptor antagonists should attenuate ET-mediated vasoconstriction. The aim of the present study was to evaluate and compare the effects of BQ-123, an ETA receptor antagonist, and bosentan, an ETA and ETB receptor antagonist, on coronary vasomotor tone, left ventricular systolic function and ET-1 efflux in the presence or absence of myocardial ischaemia/reperfusion. METHODS: Isolated rat hearts were perfused using a Langendorff preparation. Global ischaemia was induced on average by 68 +/ 2% (+/- standard error of the mean) reduction of a baseline perfusion flow-rate 10 min after introduction of ET antagonists. Thirty minutes of ischaemia was followed by 30 min reperfusion. ET-1 efflux in coronary perfusate was measured by radioimmunoassay. RESULTS: In non-ischaemic hearts (n = 7), BQ-123 (10(-6) M) perfusion induced a progressive decrease in coronary flow-rate compared with control group. This flow reduction persisted after wash-out of BQ-123. In contrast, bosentan (10(-5) M, n = 7) induced no change in perfusion rate. In the absence of ET antagonists (n = 16), there was a 22 +/- 6% post-ischaemic increase in perfusion flow-rate. BQ-123 (n = 5) but not bosentan (n = 12) abolished this post-ischaemic increase in flow-rate. Neither BQ-123 nor bosentan induced significant variation in force of contraction. In ischaemic hearts, ischaemia per se induced a transient decrease in force of contraction. Bosentan significantly (P < 0.05) accentuated and BQ-123 tended to accentuate (P = 0.06) this decrease in force of contraction during ischaemia. Bosentan but not BQ-123 significantly impaired the recovery of systolic function during reperfusion (P < 0.05). Both BQ 123 and bosentan perfusion increased ET-1 efflux rate to 730 +/- 188% and 315 +/- 81% respectively. This effect was abolished during ischaemia for BQ-123, but not for bosentan. CONCLUSIONS: In isolated perfused rat hearts, both BQ-123 and bosentan increased ET-1 efflux, but only BQ-123 exerted vasoconstrictor effects. These results thus generated the hypothesis that: (1) ET-1 release within the coronary vascular bed may be physiologically subject to negative feedback regulation mediated via ETA receptors; (2) ETA receptor antagonists increase ET-1 efflux, which may lead to net vasoconstriction via unopposed ETB stimulation. Furthermore, the negative inotropic effects observed during ischaemia suggest that ET is critical to the maintenance of systolic function during ischaemia. PMID- 9861311 TI - The endothelin A receptor antagonist LU 135252 protects the myocardium from neutrophil injury during ischaemia/reperfusion. AB - OBJECTIVE: Endothelin-1 (ET-1) is not only a potent vasoconstrictor but also a stimulator of polymorphonuclear leukocyte (PMN) aggregation and adhesion. The aim of this study was to investigate whether an ETA receptor antagonist attenuates the PMN-mediated contractile dysfunction following myocardial ischaemia. METHODS: Isolated rat hearts were perfused according to the Langendorff method. The hearts were subjected to global ischaemia and reperfused with buffer solution only, or human PMNs dissolved in rat plasma (HNRP). RESULTS: In an initial study, the ETA receptor antagonist LU 135252 (1 and 10 mumol/l) or ET-1 (1 and 10 nmol/l) did not significantly affect the recovery of left ventricular developed pressure (LVDP), end-diastolic pressure (LVEDP), the first derivative of left ventricular pressure (dP/dt) or the rate pressure product (RPP) during reperfusion with buffer solution only compared to a vehicle group. In a second study on hearts reperfused with HNRP, administration of LU 135252 (10 mumol/l) significantly enhanced the recovery of LVDP, dP/dt and RPP in hearts reperfused with HNRP. LVEDP was 20 mmHg lower in hearts given LU 135252 than vehicle in combination with HNRP (P < 0.05). The outflow of PMNs in the coronary effluent during reperfusion was 41 +/- 8% in hearts given LU 135252 compared to 9 +/- 5% in vehicle-treated hearts (P < 0.01). There was a significant correlation between the myocardial functional recovery and the outflow of PMNs. Administration of ET 1 (0.1 and 1 nmol/l) in combination with HNRP resulted in complete loss of contractile function and no outflow of PMNs during reperfusion. CONCLUSION: The ETA receptor antagonist LU 135252 protects from ischaemia/reperfusion injury in the isolated rat heart in the presence of PMNs. It is suggested that inhibition of PMN-induced injury during reperfusion is an important cardioprotective action of LU 135252. PMID- 9861312 TI - Endothelin-A and -B antagonists protect myocardial and endothelial function after ischemia/reperfusion in a rat heart transplantation model. AB - OBJECTIVE: Previous studies suggested that endothelin-1 (ET-1) may play a pathophysiological role in myocardial ischemia/reperfusion injury. This study was designed to investigate the effects of the selective ET-A receptor antagonist BQ123 and the selective ET-B receptor antagonist BQ788 on myocardial and endothelial function after reversible deep hypothermic ischemia in a heterotopic rat heart transplantation model. METHODS: Isogenic intraabdominal heterotopic transplantation was performed in Lewis rats. After 1 h of cold ischemic preservation reperfusion was started either after application of placebo (control), BQ123 (3 mumol/kg/min). BQ788 (3 mumol/kg/min), ET-1 (8 pmol/kg/min) or simultaneous infusion of BQ123 or BQ788 and ET-1, respectively (n = 12 each). An implanted balloon was used to obtain pressure-volume relations of the transplanted heart. Myocardial blood flow (MBF) was assessed by the hydrogen clearance method. Measurements were taken after 1 and 24 h of reperfusion. Endothelium-dependent vasodilation to acetylcholine (ACH) and endothelium independent vasodilation to sodium nitroprusside were also determined. RESULTS: Both BQ123 and BQ788 significantly improved myocardial and endothelial functional recovery during early reperfusion, whereas ET-1 significantly impaired myocardial and endothelial function. Simultaneous infusion of ET-1 diminished the effects of BQ123 and BQ788. Although myocardial function and baseline MBF were similar in all groups after 24 h of reperfusion, endothelium dependent vasodilation to ACH was still significantly higher in the BQ123 and BQ788 groups and lower in the ET 1 groups (p < 0.05). CONCLUSIONS: These results suggest that endogenous ET release is involved in the pathogenesis of reperfusion injury after heart transplantation. ET-A and ET-B receptor antagonists may be useful to reduce ischemia/reperfusion injury. PMID- 9861313 TI - Controversies in the management of Graves' disease. PMID- 9861314 TI - Growth hormone and cortisol metabolism. PMID- 9861315 TI - Short Synacthen tests--the need for rationalization. PMID- 9861316 TI - Too much of a good thing. PMID- 9861317 TI - Defining the normal cortisol response to the short Synacthen test: implications for the investigation of hypothalamic-pituitary disorders. AB - OBJECTIVE: To define the normal cortisol response to the Short Synacthen Test using four different cortisol immunoassays and to assess the implications for the investigation of hypothalamic-pituitary disorders. DESIGN AND PATIENTS: The cortisol response to 250 micrograms im ACTH1-24 (Synacthen, Ciba Geigy) in 100 healthy volunteers using four different cortisol immunoassays has been measured. In 44 newly diagnosed and untreated patients with pituitary disease, basal and 30 minute post-ACTH cortisol results were also determined using the four immunoassays. RESULTS: The distribution of cortisol results at all time points and for all methods were non-Gaussian and significant differences in the absolute values of the 5th-95th percentiles were found between methods (P < 0.01). At 30 min post-Synacthen in normals the 5th percentile of the cortisol response ranged from 510 to 626 nmol/l with the different methods. Similarly the relationship between assay results differed at different time points. No effect of age on the cortisol response was found but for stimulated cortisol values and the incremental responses females showed significantly higher responses than males (P < 0.05) for most methods. Although there was a significant positive linear correlation (P < 0.001) between stimulated and basal cortisol values for all methods, no significant relationship was found between the incremental response and basal cortisol values. In the pituitary disease patients basal and 30 minute post-ACTH cortisol results were significantly lower (P < 0.05 and < 0.001) than the control group using the same cortisol assay. When the results were compared to the 5th percentile of the gender and assay specific control group 33.3% of male and 17.4% of female patients failed the Synacthen test at 30 min. CONCLUSIONS: The definition of the 'normal' response to Synacthen should be both gender and method related at all time points. The data suggest that up to one third of untreated patients with pituitary disease may have subtle defects in the hypothalamic-pituitary-adrenal axis. PMID- 9861318 TI - Comparative evaluation of conventional and dynamic magnetic resonance imaging of the pituitary gland for the diagnosis of Cushing's disease. AB - OBJECTIVES: The ability of MRI to detect pituitary ACTH-secreting adenomas in patients with Cushing's disease is limited. Owing to different dynamics of contrast enhancement between adenomas and normal pituitary tissue, it has been suggested that obtaining images within seconds after gadolinium (Gad) injection using dynamic procedures increases the sensitivity of MRI in the detection of pituitary microadenomas. The objective of this study was to compare the ability of conventional magnetic resonance imaging (CMRI) and dynamic MRI (DMRI) to detect ACTH-secreting pituitary adenomas. DESIGN: Twenty-six consecutive patients with ACTH-dependent Cushing's syndrome and 10 normal subjects were investigated. According to the results of inferior petrosal sinus sampling, 21 patients had Cushing's disease and five had ectopic ACTH syndrome. Patients with Cushing's disease were operated regardless of the results of imaging studies. All underwent identical MRI and DMRI procedures using a 1.0 T magnet. Image sampling time during DMRI was 19 sec. Scans were randomly mixed and analysed blind, retrospectively and independently by two experienced radiologists. The clarity of the images was assessed by the analysis of agreement among radiologists. MRI findings were compared to surgical and histopathological findings. RESULTS: Surgical exploration identified three macrodenomas and 14 microadenomas. One microadenoma was found at pathological examination after subtotal hypophysectomy and no tumour was found in three cases. According to the combined opinion of radiologists, the three macroadenomas were identified equally well with CMRI and DMRI. Eight ACTH-secreting microadenomas were detected with CMRI and 11 with DMRI. The three microadenomas detected with DMRI only were visualized within 60 sec following Gad injection. No false positives occurred with CMRI. Three false positives were obtained with DMRI: one in a patient with ectopic ACTH syndrome while a silent microprolactinoma and normal tissue were found at the site of the radiological abnormality in two patients with Cushing's disease. In our study, the sensitivity of DMRI is greater than that of CMRI (0.67 vs. 0.52) but is associated with a loss in specificity (0.80 vs. 1.00). False positives may result from the increased sensitivity of DMRI which detects incidental pituitary lesions, technical artefacts or lowest clarity of images, as suggested by a lower observer agreement of DMRI (Kappa statistic 0.66 vs. 0.83). Overall, the two MR procedures had equivalent diagnostic power (0.72). CONCLUSIONS: In our hands, dynamic procedures did not improve the usefulness of MRI in Cushing's syndrome. PMID- 9861319 TI - Low level of glucocorticoid receptor messenger ribonucleic acid in pituitary adenomas manifesting Cushing's disease with resistance to a high dose dexamethasone suppression test. AB - OBJECTIVES: The overnight 8-mg dexamethasone suppression test is often used to differentiate Cushing's disease, due to an oversecretion of ACTH from the pituitary gland, from other kinds of Cushing's syndrome. However, a few patients with ACTH-producing pituitary adenoma show no suppression of plasma cortisol after the administration of 8 mg of dexamethasone. To clarify the relationship between the level of glucocorticoid receptor (GR) in the pituitary adenoma and the sensitivity to dexamethasone in Cushing's disease, we thus examined the levels of GR alpha and GR beta mRNAs in the pituitary adenomas in six patients who were proven at surgery to have pituitary ACTH-producing adenomas. MATERIALS: Total RNA was extracted from six pituitary adenomas and pituitary tissue adjacent to one of the adenomas, and the mRNA levels of GR alpha, GR beta, pro opiomelanocortin (POMC) and beta-actin in these samples were sampled by quantitative RT-PCR. RESULTS: The GR alpha mRNA levels in the adenomas from the two patients who showed no response to the 8-mg dexamethasone suppression test were significantly lower than those in the adenomas of four patients who showed suppression. The GR beta mRNA level was much lower than that of GR alpha mRNA but not significantly different among the six adenomas. CONCLUSIONS: These results suggest strongly that decreased expression of GR alpha in pituitary adenomas may be the major reason for the marked insensitivity to the 8-mg dexamethasone suppression test observed in two patients with Cushing's disease. PMID- 9861320 TI - Reproducibility of the low dose dexamethasone suppression test: comparison between direct plasma and salivary cortisol assays. AB - BACKGROUND: The low dose dexamethasone suppression test (DST) has been used to detect subtle variations in the feedback suppression of the hypothalamic pituitary-adrenal axis, which may contribute to the pathogenesis of several diseases including depression, the metabolic syndrome and coronary artery disease. Little is known about the reproducibility of this test, or whether the test can be combined with analysis of salivary cortisol which would offer a significant advantage over plasma in population studies. SUBJECTS AND DESIGN: A low dose DST was carried out in 29 healthy subjects (14 men, 15 women), aged 24 54 (mean 35.1) years, on two separate occasions 1-10 weeks apart. Following the administration of 0.25 mg dexamethasone (DXM) at 2200 h, plasma and saliva were sampled at 0830 h the next day. Cortisol was measured by radioimmunoassay in plasma and time-resolved immunofluorescent assay ('DELFIA') in saliva. Bland Altman plots were produced for post-DXM plasma and salivary cortisol measures and used to derive a coefficient of repeatability for each measure, which describes the range of cortisol measurements within which 95% of repeated measurements will fall. RESULTS: The baseline, pre-DXM cortisol concentrations were far more variable for saliva (mean 16.5, range 4.4-34 nmol/l) than for plasma (mean 407.5; range 232-958 nmol/l). Following DXM both measurements showed an approximately 30% suppression from baseline but the variability of salivary cortisol was much greater. From the Bland-Altman plots the 95% range for the differences about their mean was calculated and used as an indication of repeatability. For plasma 95% of differences were within 0.78 log units, indicating that a repeated measurement was approximately half as small or twice as large as the first. For saliva 95% of differences were within 1.64, indicating that a repeated measurement was approximately five times as small or five times as large as the first. CONCLUSIONS: Assessment of dexamethasone suppression by salivary cortisol measurement is far less repeatable than the use of plasma cortisol. In the context of field studies of dexamethasone suppression, salivary cortisol measurements may only be appropriate for large numbers of subjects. PMID- 9861321 TI - Age-dependent decline in cortisol levels and clinical manifestations in patients with ACTH-independent Cushing's syndrome. AB - OBJECTIVE: Adrenocortical cells change with age in both subcellular morphology and level of steroidogenesis. This study evaluates the influence of aging on hypercortisolism and clinical manifestations in patients with ACTH-independent Cushing's syndrome due to a cortisol-secreting adrenal adenoma (CS). DESIGN: Restrospective study. PATIENTS: Thirty-six (33 females and 3 males) with CS. 31 healthy controls were age- and sex-matched to CS patients. MEASUREMENT: Patient age at diagnosis was compared to the degree of hypercortisolism and clinical manifestations. The degree of hypercortisolism was estimated using serum cortisol levels and urinary free cortisol excretion. Positive clinical manifestations were quantified using 12 symptoms and signs which resulted in a clinical score for each patient. Endogenous creatinine clearance, estimated disease duration and gender were analysed as possible factors affecting adenoma-secreated cortisol. RESULTS: Patient age correlated negatively with serum cortisol level (r = -0.417, P = 0.0107, n = 36) and urinary excretion of free cortisol (r = -0.613, P = 0.0002, n = 31). Analysis of Spearman's rank correlation showed that the clinical score decreased with age (rho = -0.631, P = 0.0004, n = 33). Oedema, weakness/myopathy, hirsutism, striae and psychological changes were infrequent symptoms in the elderly. Using stepwise multiple regression analysis, we estimated the clinical score (Y) by factors of serum cortisol levels (X1:nmol/l), age (X2: years of age) and gender (X3: gender; female = 1, male = 0), i.e. Y = 4.771 + 0.004 X1 - 0.064 X2 + 2.548 X3 (r = 0.743, P < 0.0001, n = 33). Results suggested that aging both suppressed cortisol secretion and repressed manifestations directly as an independent factor, although its direct effect may be weak. CONCLUSIONS: Our findings indicated the importance of age as a factor modulating hormonal levels and possible clinical manifestations in patients with cortisol-secreting adenomas. PMID- 9861322 TI - Prevalences of Gs alpha, ras, p53 mutations and ret/PTC rearrangement in differentiated thyroid tumours in a Korean population. AB - OBJECTIVE: In thyroid tumours, ras, Gs alpha, p53 mutations and ret/PTC rearrangement have been reported with variable prevalences in different geographical regions. We studied the prevalence of these mutations and rearrangement in thyroid tumours in a Korean population. As MDM2 and Bcl-1 protein expressions have been suggested to be associated with p53 protein, we also studied possible relationships among them. PATIENTS AND DESIGN: Eleven cases of adenomatous goitre, eight cases of follicular adenoma, five cases of follicular carcinoma and 37 cases of papillary carcinoma were included in this study. To find mutations and rearrangement, RT-PCR, SSCP and/or direct sequencing, after subcloning if necessary, were used, and immunohistochemical stainings were performed for p53, MDM2 and Bcl-2 proteins in cases of papillary carcinoma. RESULTS: We could not find any rearrangement for ret/PTC-1, -2, -3 and mutation of Gs alpha. For the ras oncogene, K and H-ras mutations were not found, but N-ras mutations, point mutation of CAA to CGA in codon 61, were detected in one follicular adenoma (12.5%, 1/8) and one follicular carcinoma (33%, 1/3). p53 mutations were detected in only one case of papillary carcinoma (3%, 1/31: exon 8, codon 266 GGA-->GAA). In 30 cases of papillary carcinoma without p53 mutation, the prevalences of positive immunohistochemical staining were 13.3% for p53 protein, 53.3% for MDM2 protein and 56.7% for Bcl-2 protein. While over expression of p53 protein was not significantly related to that of MDM2 and Bcl-2 proteins, over-expression of MDM2 and Bcl-2 in papillary carcinoma were associated. CONCLUSION: ret/PTC rearrangement, Gs alpha, ras and p53 mutations are relatively rare in differentiated thyroid neoplasms from a Korean population, which may reflect genetic and environmental differences from patients in countries with high prevalences. P53 protein over-expression was noted in 13.3% of papillary carcinoma cases without p53 mutation and was not significantly related to MDM2 and Bcl-2 expression. PMID- 9861323 TI - Gestational transient hyperthyroxinaemia (GTH): screening for thyroid function in 23,163 pregnant women using dried blood spots. AB - OBJECTIVE: Transient elevation of serum free T4 (gestational transient hyperthyroxinaemia; GTH) occurs occasionally during normal pregnancy, especially in early gestation. However, the frequency of GTH and its clinical features remain unclear to date. The aim of this study was to determine the occurrence rate of GTH and the relation between serum levels of hCG, free T4 (fT4), and TSH in a large number of pregnant women. DESIGN: The four criteria of GTH were as follows: (1) no past history of thyroid disease, (2) negative tests for MCHA and TGHA, (3) no multiple pregnancies or trophoblastic disease and (4) transient hyperthyroxinaemia at less than 16 weeks of gestation. Thyroid function and hCG levels in 23,163 pregnant women were evaluated by mass sreening. If individual fT4 levels were more than the upper limit, blood re-sampling and the clinical and laboratory analysis of thyroid function were performed to exclude women with thyroid disease. The concentrations of hCG, fT4, and TSH in women with GTH and normal pregnant controls (n = 218) were compared. Regression analysis was performed for the comparison between hCG, fT4, and TSH levels in women with GTH. MEASUREMENTS: Blood samples were obtained using dried blood spots. Blood levels of fT4 was measured by radioimmunoassay, TSH and hCG were measured by fluoroimmunoassay. Anti-microsome antibody (MCHA) and anti-thyroglobulin antibody (TGHA) were measured by indirect agglutination reaction. RESULTS: GTH was observed in 66 of 23,163 women. The overall occurrence rate of GTH was 0.285%. In 22 of the 66 GTH women, serum TSH was undetectable. Using regression analyses, the concentration of fT4 was correlated with hCG levels in women with GTH (P < 0.05, r = 0.269), whereas the concentration of TSH was not correlated with hCG or fT4 level. The concentrations (M +/- SD) of fT4, TSH, and hCG in women with GTH were 42.5 +/- 12.3 pmol/l, 0.20 +/- 0.31 mU/l and 190.2 +/- 98.8 x 10(3) IU/l, whereas those of controls were 14.6 +/- 3.8 pmol/l, 1.43 +/- 1.25 mU/l and 60.1 +/- 45.1 x 10(3) IU/l. The concentrations of fT4 and hCG were significantly (P < 0.0001) higher than those of normal controls, and TSH was significantly (P < 0.0001) lower than those of normal controls. CONCLUSION: The occurrence rate of gestational transient hyperthyroxinaemia was 0.285%, and could possibly be attributed to increased levels of circulating hCG. Based on the data obtained from a large number of pregnant women, we propose gestational transient hyperthyroxinaemia as a definite clinical entity. PMID- 9861324 TI - No evidence for allelic association of a human CTLA-4 promoter polymorphism with autoimmune thyroid disease in either population-based case-control or family based studies. AB - OBJECTIVE: The cytotoxic T lymphocyte associated-4 (CTLA-4) gene is a candidate for T-cell mediated autoimmune disease and polymorphism has been reported to be associated with both type 1 diabetes and autoimmune thyroid disease. A previously unreported polymorphism of the promoter region of the human CTLA-4 gene has recently been described in a sample of a normal control population. We investigated the distribution of this polymorphism, situated at position -318 to the ATG start codon and resulting in a C-T change leading to an Mse I restriction site, in both population based case control studies and family studies in patients with Graves' disease (Caucasian and Hong Kong Chinese), autoimmune hypothyroidism and systemic lupus erythematosus (SLE). DESIGN: Target DNA was amplified using the polymerase chain reaction and the resulting product was digested using the Mse I restriction enzyme. PATIENTS: One hundred and ninety-one white UK Caucasian and 98 Hong Kong Chinese patients with Graves' disease, 78 white UK Caucasian patients with Graves' disease plus family members, 92 white UK Caucasian patients with autoimmune hypothyroidism, 13 white UK Caucasian patients with autoimmune hypothyroidism plus family members, 132 white UK Caucasian patients with systemic lupus erythematosus, 355 white UK Caucasian control subjects and 82 Hong Kong Chinese control subjects. MEASUREMENTS: Frequencies of the C and T alleles were compared between patients and control subjects using the chi 2-test and Fisher's exact test for small numbers. RESULTS: No association with the T allele was observed in any of the patient groups studied. CONCLUSION: These data suggest that the C-T change in exon 1 of the promoter region of the CTLA-4 gene does not play a role, nor is in linkage disequilibrium with a disease causing mutation, in the development of autoimmune disease. PMID- 9861325 TI - The systemic stress response to thermal injury in children. AB - OBJECTIVE: Thermal injury is extremely stressful, but data characterizing the endocrine stress response to this injury in children are sparse. The objective of this study was to measure the effects of thermal injury on the levels of stress hormones in children and to assess the temporal changes associated with them. PATIENTS: Twenty-three children, 13 girls and 10 boys aged between 5 months and 12 years 3 months (mean, 2 years 11 months), with burns covering 10-61% of their body surface (mean, 20.5%) were studied during the first 5 days following injury. MEASUREMENTS: The levels of arginine vasopressin, angiotensin II, cortisol, adrenaline, noradrenaline and dopamine were measured in sequential blood samples obtained from thermally injured children on admission and at specified time intervals during the 5 days of the investigation. RESULTS: At admission the concentrations of all the hormones were high, and varied widely between individual patients. The geometric mean and 95% confidence intervals of admission hormone levels were as follows: arginine vasopressin 18.3 (8.3-40.7) pmol/l; angiotensin II 122.0 (56.0-266.2) pmol/l; cortisol 650.6 (473.0-895.0) nmol/l; dopamine 1.0 (0.1-8.0) nmol/l; adrenaline 6.4 (3.2-12.5) nmol/l and noradrenaline 2.3 (1.3-4.3) nmol/l. Although the concentrations of arginine vasopressin and cortisol returned to normal 24 to 36 h after admission, the levels of angiotensin II, adrenaline and dopamine fluctuated and remained higher than normal throughout the study (108 h). CONCLUSIONS: Thermal injury results in the release of abnormally high levels of stress hormones in children. Although there are similarities between some of the data reported here and those reported in adults, higher levels of adrenaline and lower levels of noradrenaline than reported in adults suggest important differences too. These differences may need to be taken into account in the management of burn-injured children. PMID- 9861326 TI - Effects of chronic opioid antagonism on gonadotrophin and ovarian sex steroid secretion during the luteal phase. AB - OBJECTIVES: Since short-term opioid antagonism increases LH pulsatility during the luteal phase in women, we postulated that prolonged opioid antagonism may also accelerate the LH secretory episodes at this time. If so, the functional and temporal links between secretory episodes of pituitary LH and oestradiol (E2) and progesterone (P) release from the mature human corpus luteum may be disrupted. STUDY DESIGN: Prolonged opioid blockade with the oral antagonist naltrexone (100 mg daily) was effected in eight women during the entire luteal phase of their cycles. Following documented ovulation in both placebo (control) and naltrexone cycles, blood samples were obtained daily and frequently (every 10 minutes for 10 h) on days 6-8 after ovulation. MEASUREMENTS: In all blood samples, LH, E2 and P were determined by IRMA. RESULTS: Compared to control cycles, the temporal organization and the endocrine characteristics of the luteal phase remained virtually unchanged during chronic opioid blockade. Periodic fluctuations were detected (by cluster analysis) in LH, E2 and P data series established by frequent sample collections in both the control and naltrexone cycles. LH secretory profiles were remarkably similar during control and naltrexone cycles, and the E2 and P secretory episodes tended to be coupled to LH pulses during both cycles. As determined by time-series analysis, the cross-correlations between the LH/E2 and LH/P data series remained unaltered by opioid blockade. CONCLUSIONS: Chronic opioid antagonism with naltrexone did not disrupt the temporal organization or endocrine characteristics of the luteal phase. In particular, prolonged opioid blockade did not change LH secretory patterns. The functional and temporal links between LH inputs and sex steroid release from the mature corpus luteum remained unaffected by prolonged opioid antagonism. In contrast to the effects of short-term opioid blockade on LH pulsatile release during the luteal phase, the effects of chronic opioid antagonism on LH release may be transient and may not persist throughout the entire luteal phase, suggesting desensitization of the opiate receptors. PMID- 9861327 TI - Adrenal steroid hormones in short children born small for gestational age. AB - OBJECTIVE: Programming of the endocrine axis has been postulated to occur during critical phases of fetal development and is affected by intrauterine growth retardation. The aim of this study was to investigate this hypothesis with regard to adrenal steroid hormones. Thus, serum cortisol and dehydroepiandrosterone sulphate (DHEAS) levels were compared in children born small for gestational age (SGA) who remained short and in children born at an appropriate size for gestational age (AGA), of both short and normal stature. DESIGN AND PATIENTS: Seven serum samples for cortisol measurements were taken during a 24-h period from a total of 184 prepubertal individuals. The study group comprised 53 children born SGA who remained short (41 boys, 12 girls; mean chronological age, 8.8 +/- 2.5 years). The reference groups of children born AGA were as follows: 75 healthy short children (56 boys, 19 girls; mean chronological age, 10.8 +/- 2.6 years) and 56 healthy children of normal height (37 boys, 19 girls; mean chronological age, 11.3 +/- 1.8 years). A single serum sample for measurement of DHEAS was taken between 1000 and 1400 h in 110 of the 184 children (33 short SGA, 42 short AGA and 35 AGA of normal height). MEASUREMENTS: Serum cortisol and DHEAS were measured by radioimmunoassay. RESULTS: No differences were found between children born SGA and children born AGA in either cortisol levels, calculated as area under the curve (AUC), or the circadian cortisol rhythm, estimated from the calculated nadir, the peak and the amplitude. No difference between the groups was found for serum DHEAS concentrations. Serum cortisol levels, expressed as AUC, and serum DHEAS levels did not correlate with size at birth. However, when adjusted for age at investigation, serum DHEAS, but not serum cortisol, correlated with weight at birth. CONCLUSIONS: Serum cortisol levels and rhythms do not correlate with size at birth and are similar in children born small for gestational age who remain short and children born appropriate size for gestational age of both short and normal stature. However, DHEAS levels in young children before adrenarche correlated inversely with weight at birth, indicating a relationship with fetal growth. PMID- 9861328 TI - Gonadotrophin and prolactin secretory dynamics in girls with normal puberty, idiopathic precocious puberty and precocious puberty due to hypothalamic hamartoma. AB - OBJECTIVE: This study was designed to test the hypothesis that hypothalamic hamartoma causes precocious puberty through a different neuroendocrine mechanism than that of normal puberty or of idiopathic precocious puberty. DESIGN AND PATIENTS: We compared the pattern of gonadotrophin secretion among 4 girls with precocious puberty due to hypothalamic hamartoma, 27 girls with idiopathic precocious puberty, and 14 girls with normal puberty. All subjects were breast stage 3 or 4. Blood samples were obtained every 20 min for 4 h during the day (1.000 hours to 1400 h) and night (22.00 hours to 0200 h). MEASUREMENTS: LH, FSH, and prolactin were measured in each blood sample. Girls also underwent LHRH stimulation with measurement of LH and FSH before and after stimulation. RESULTS: There were no significant differences in mean LH level, LH peak amplitude, or LH or FSH peak frequency during either the day or the night among the three diagnostic groups. However, the mean +/- SD LHRH-stimulated peak LH levels were greater in girls with hypothalamic hamartoma than in girls with normal puberty or with idiopathic precocious puberty (194 +/- 142 vs 85 +/- 60 or 66 +/- 54 IU/l, respectively, P < 0.05). The LHRH-stimulated peak FSH level in girls with hypothalamic hamartoma exceeded the level for the normal pubertal girls (31 +/- 19 vs 17 +/- 7 IU/l, P < 0.05), but not the level for the girls with idiopathic precocious puberty (25 + 12 IU/l). The peak LH to peak FSH ratio in the girls with hypothalamic hamartoma exceeded the ratio for the girls with idiopathic precocious puberty (7.3 +/- 3.9 vs 2.6 +/- 3.0 IU/l, P < 0.05), but not the ratio for the normal pubertal girls (5.0 + 2.9). There were no significant differences in mean prolactin level, peak amplitude or frequency, or in the ratio of mean night to mean day prolactin, among the 3 diagnostic groups. CONCLUSIONS: We conclude that spontaneous gonadotrophin and prolactin secretion are similar among girls with hypothalamic hamartoma, idiopathic precocious puberty, or normal puberty. However, the increased LHRH-stimulated peak LH in the girls with hypothalamic hamartoma suggests subtle differences in neuroendocrine regulation that may underlie their more rapid pubertal maturation. PMID- 9861329 TI - Determination of a common genetic variant of luteinizing hormone using DNA hybridization and immunoassays. AB - OBJECTIVE: An immunologically anomalous form of LH, due to two point mutations in codons 8 and 15 of the LH beta gene, has previously been described. LH status, i.e. the discrimination between wild-type (WT) and variant (V) LH, is usually determined by immunoassays, which can be unreliable at low serum concentrations of LH. A DNA hybridization assay was therefore developed to score the LH genotype in all subjects, independent of their serum LH concentrations. To evaluate the performance of the hybridization method, and to expand our observations of the worldwide occurrence of the V-LH, we determined its frequency in additional populations. To confirm the connection between the anomalous immunoreactivity and the V-LH beta gene, we also sequenced the LH beta subunit gene of a homozygous person. DESIGN: According to the ratio of two immunoassays, one detecting only WT LH and the other detecting equally WT and V-LH, individuals can be classified as homozygotes for the V-LH beta allele, heterozygotes or WT. DNA samples from persons with known LH status, according to the immunoassays, were used for the development and evaluation of a new allele-specific DNA hybridization assay. This assay, and PCR and restriction fragment length polymorphism analysis, were used to determine the frequency of the V-LH beta allele in DNA samples obtained from eight populations. PATIENTS: Ambulatory adult men and women, apparently healthy and with no endocrine disorders. RESULTS: The LH genotyping by immunoassays and by the new hybridization method gave identical results with all samples analysed (n = 25). The V-LH beta subunit was observed to always have the two point mutations, and to be identical with the ones previously reported. The V-LH beta carrier frequency in the DNA samples collected from various populations varied between 0 and 53.5%. CONCLUSIONS: The immunoassay technique and the hybridization assay can be used as alternatives to determine the LH status. A great variation in carrier frequency of the V-LH beta allele is observed in different populations. PMID- 9861330 TI - Reduced serum dehydroepiandrosterone levels in diabetic patients with hyperinsulinaemia. AB - OBJECTIVE: To elucidate the interaction between insulin and dehydroepi androsterone (DHEA) concentrations, we evaluated serum DHEA and DHEA-sulphate (DHEA-S) levels in diabetic patients with hyperinsulinaemia. PATIENTS AND DESIGN: Twenty-four subjects with non-insulin dependent diabetes mellitus, 12 hyperinsulinaemic subjects (fasting serum insulin concentrations > or = 10 mU/ml (71.8 pmol/l)) and 12 non-hyperinsulinaemic subjects, and 10 normal control subjects were studied. Serum DHEA, DHEA-S, cortisol and ACTH levels were investigated in these subjects. Moreover, their serum DHEA levels were compared during hyperinsulinaemic-euglycaemic clamp and after ACTH stimulation. MEASUREMENTS: Serum insulin, cortisol, ACTH, DHEA and DHEA-S concentrations were evaluated by RIA. Serum glucose was determined by the glucose oxidase method. RESULTS: Diabetic patients with hyperinsulinaemia showed significantly lower levels of serum DHEA and DHEA-S than controls. After ACTH stimulation, these patients also showed significantly lower DHEA levels. During the hyperinsulinaemic-euglycaemic clamp, serum DHEA concentrations of diabetic patients with hyperinsulinaemia remained low and did not decline further, although those of control subjects and non-hyperinsulinaemic diabetic patients showed a significant decline of serum DHEA levels. Even after ACTH stimulation during the clamp, serum DHEA in hyperinsulinaemic patients was still significantly lower than in controls. CONCLUSIONS: In diabetic patients with hyperinsulinaemia, baseline DHEA levels are chronically and maximally suppressed compared to control subjects and non-hyperinsulinaemic diabetic patients, and thus not decreased further by exogenous insulin infusion during hyperinsulinaemic euglycaemic clamp. PMID- 9861333 TI - Bone mineral density in women with cytotoxic-induced ovarian failure. AB - OBJECTIVE: Premature ovarian failure is associated with a reduction in bone mineral density. As survival rates following treatment for haematological malignancies improve, chemotherapy-induced ovarian failure is becoming more common. However, there are few data concerning the impact of this on bone mineral density (BMD). We have therefore measured the BMD in 33 women with ovarian failure following treatment with cytotoxic chemotherapy. PATIENTS AND DESIGN: We studied 33 women who received combination chemotherapy for Hodgkin's disease (n = 27), non-Hodgkin's lymphoma (n = 4), sarcoma (n = 1) and acute myeloid leukaemia (n = 1). The mean (range) age of the subjects at the time of BMD measurement was 37.5 (24-50) years and the mean (median: range) duration of amenorrhoea was 49 (24: 5-277) months. Eleven women had received hormone replacement therapy (HRT) for a mean (range) duration of 25 (1-62) months. BMD was measured by single photon absorptiometry or single X-ray absorptiometry, and dual energy X-ray absorptiometry at the distal and proximal radius, the femoral neck and the lumbar spine, respectively. BMD was expressed as Z-scores and statistical analysis was performed using the Wilcoxon matched-pairs signed-rank test. RESULTS: There was no significant reduction in BMD at the hip, spine or a forearm in the cohort as a whole, although there was a trend to reduce bone density at all sites. When patients who had received HRT were excluded from the analysis there were small reductions in mean BMD at all sites, but this was only statistically significant at the proximal forearm (Z-score = -0.65; P = 0.03). Mean BMD of the HRT-treated patients was normal at all sites. Only seven patients (21%) had a BMD Z-score < 2 at any site. CONCLUSION: It is inappropriate to assume that ovarian failure from different aetiologies has a similar deleterious impact on the skeleton. Untreated premature ovarian failure following cytotoxic chemotherapy results in some reduction in bone mineral density, but this is of a minor degree and is less than that observed in other hypo-oestrogenic states. The reason for this is unclear but studies of residual hormone production in the cytotoxic-damaged ovary may provide an answer. PMID- 9861332 TI - Cognitive dysfunction in patients with pituitary tumour who have been treated with transfrontal or transsphenoidal surgery or medication. AB - OBJECTIVE: This study was carried out to examine the neuropsychological status of patients treated for pituitary tumour by transfrontal surgery, transphenoidal surgery or medical treatment only, with or without radiotherapy. DESIGN AND MEASUREMENTS: Three groups of 23 patients who had been treated for pituitary tumour were compared with 23 healthy controls on a range of neuropsychological measures. The surgical patients were also subdivided into two groups and compared. The neuropsychological measures were standardized psychological tests designed to assess aspects of attention, memory and executive function. PATIENTS: The patients were those who had been treated with transfrontal surgery (n = 23), transsphenoidal surgery (n = 23) and medication only (n = 23). The groups did not differ with respect to age, education or premorbid ability level as assessed by the National Adult Reading Test. All participants were free of known sources of cognitive impairment other than pituitary tumour. RESULTS: Comparison of the four groups revealed that nearly half of the transfrontal, one-third of the transsphenoidal and one-quarter of the non-surgical group had three or more neuropsychological tests scores below the 10th percentile compared to less than 5% of the controls. Impairments in memory and executive function were found in both surgical groups. The non-surgical patients appeared to have problems only on tasks requiring high levels of cognitive processing. Differences were found between the two surgical groups with respect to the severity of the cognitive impairment, the transfrontal patients having more severe impairment than the transsphenoidal. No significant negative effects on cognitive functioning were associated with radiotherapy; however, transfrontal surgery patients who had not been treated with radiotherapy were found to be more impaired than other patients. This was thought to be related to radical surgery. CONCLUSIONS: Many patients with treated pituitary tumour suffer significant cognitive impairment. The severity and nature of impairment differs between treatment groups, although the cause of this could not be addressed by this study. Recommendations are made for future research and clinical practice. PMID- 9861331 TI - Serum leptin and insulin concentrations in prepubertal lean, obese and insulin dependent diabetes mellitus children. AB - OBJECTIVE: To evaluate the relationship between serum levels of leptin and insulin in prepubertal lean, obese and insulin-dependent diabetes mellitus (IDDM) children. SUBJECTS AND MEASUREMENTS: Prepubertal children, 16 lean, 17 obese and 16 IDDM were included in the study. Fastang serum leptin and insulin concentrations were measured by radioimmunoassays. RESULTS: The serum level of leptin was significantly higher in obese children than in lean and IDDM children (P < 0.0001 and P < 0.0001, respectively), and showed a positive correlation with body mass index (BMI) for the combined group (lean, obese and IDDM; r = 0.77, P < 0.0001). In addition, the serum leptin level was higher in IDDM children than in lean controls (P < 0.01), whereas no difference was found in BMI between the two groups. The mean fasting serum levels of insulin were significantly elevated in IDDM children as compared with lean controls (P < 0.01). A significant positive correlation was found between serum insulin and leptin levels for the combined group (r = 0.37, P < 0.01). When a multiple regression analysis for all subjects was performed, the total contribution of all parameters, including gender, BMI and log insulin, accounted for 75% of the leptin variation. BMI (57.8%), log insulin (14.0%) and gender (3.2%) contributed significantly to this variation. CONCLUSIONS: The elevated concentration of leptin in insulin-dependent diabetic children, independent of body mass index, was probably caused by chronically increased serum insulin levels. We demonstrated that not only body mass index but also insulin was a significant independent predictor of serum leptin concentrations. It is therefore suggested that insulin might play an important role in regulating serum leptin concentrations independent of adiposity. PMID- 9861335 TI - Unicystic ameloblastoma. A review of 193 cases from the literature. AB - Based on a world-wide literature survey of 193 published cases of unicystic ameloblastomas (UA), data have been produced allowing the presentation of a revised concept of this much debated lesion. UA is a variant of the solid or multicystic ameloblastoma. Radiographically, the unilocular pattern is more common that the multilocular, especially in cases associated with tooth impaction. However, it is stressed that although the lesion is pathomorphologically unicystic, it will far from always produce a unilocular radiolucency. The mean age at the time of diagnosis of UA is closely related to an association with an impacted tooth. Almost 20 years separate the mean age of the 'dentigerous' variant from the 'non-dentigerous' (16.5 years versus 35.2 years) The male:female ratio for the 'dentigerous' type is 1.5:1, but for the 'non-dentigerous' type it is reversed (1:1.8). Location favours greatly the mandible (mandible to maxilla = 3 to 13:1). Between 50 and 80% of cases are associated with tooth impaction, the mandibular third molar being most often involved. The 'dentigerous' type occurs on average 8 years earlier than the 'non dentigerious' variant. The mean age for unilocular, impaction-associated UAs is 22 years, whereas the mean age for the multilocular lesion unrelated to an impacted tooth is 33 years. Histologically, the minimum criterion for diagnosing a lesion as UA is the demonstration of a single cystic sac lined by odontogenic (ameloblastomatous) epithelium often seen only in focal areas. This simple type of UA (according to the authors' modification of the classification by Ackermann et al. (Journal of Oral Pathology 1988; 17:541-546)), is one of four UA subtypes, the others being (1) simple with intralumenal proliferations; (2) simple with both intralumenal and intramural proliferations; and (3) simple with intramural proliferations only. All four subtypes occur in both the 'dentigerous' and 'non dentigerous' variants. The simple subtype with and without intralumenal proliferations may be treated conservatively (enucleation), whereas subtypes showing intramural growths must be treated radically, i.e. as a solid or multicystic ameloblastoma. Finally, the authors disclose areas and issues pertaining to UA that still need to be addressed. PMID- 9861334 TI - In vivo and in vitro effects of AVP and V1a receptor antagonist on Cushing's syndrome due to ACTH-independent bilateral macronodular adrenocortical hyperplasia. AB - We examined the possibility that AVP and V1a receptors were involved in regulating cortisol production in a 49 year old man with ACTH-independent bilateral macronodular adrenocortical hyperplasia (AIMAH), and investigated the effects of a V1a receptor antagonist. An i.v. injection of a small dose (0.1 or 0.3 U) of AVP, insulin-induced hypoglycaemia, upright posture tests, and oral administration of a V1a receptor antagonist (OPC-21268; 300 mg), and its repeated administration at a dose of 600 mg/day for 8 days were performed. An in vitro study of dispersed cells obtained from resected AIMAH tissue was also conducted. Plasma ACTH, AVP and cortisol levels and 24-h urinary free cortisol excretion were measured in the in vivo studies and cortisol concentrations in incubation media in the in vitro study. Injection of small doses of AVP stimulated cortisol secretion without any elevation of plasma ACTH. Insulin-induced hypoglycaemia caused a rise in plasma AVP followed by an increase in plasma cortisol. Although plasma cortisol levels were not affected by single or repeated administrations of OPC-21268, 24-h urinary free cortisol excretion was significantly decreased by the repeated treatment. In the in vitro study, more cortisol was stimulated by AVP from adrenal cells of the AIMAH tissue than from those of a normal adrenal gland, and this secretion was completely suppressed by OPC-21268. These results suggested that hypersensitivity to AVP may have contributed to overproduction of cortisol in this case of ACTH-independent bilateral macronodular adrenocortical hyperplasia, and may have contributed to its pathogenesis. PMID- 9861336 TI - Effects of the anticancer drug cocktail "UFT" on metastatic potentials of squamous cell carcinoma (O-1N) in a hamster model. AB - UFT, a mixture of 1-(2-tetrahydrofuryl)-5-fluorouracil (tegafur) and uracil, is one of the most widely used anticancer agents. The tissue localization and the efficacy of UFT in preventing lymph node metastasis were studied in hamsters transplanted with oral squamous cell carcinoma (termed "O-1N") which has high lymph node metastatic potency. After UFT administration for 14 consecutive days, the incidence of lymph node metastasis was significantly decreased (16% vs 58%). When UFT was administered orally for 3 consecutive days to hamsters bearing O-1N, and tissue concentrations of tegafur and 5-FU were measured by gas chromatography, the value of 5-FU was significantly higher in the primary transplant tumour, cervical lymph nodes with or without metastasis, and liver, than in the normal oral tissues. When 14C-UFT(14C-tegafur + uracil, a uracil/FT molar ratio of 4) was administered orally for 3 consecutive days to hamsters bearing O-1N, the 14C-tegafur level of tissue homogenates in the buccal and metastatic tumours was within the range of normal oral tissues, but the localization of silver grains was significantly higher in tumour cells than in the oral normal tissues. The results suggest that UFT would be useful for treatment and prevention of lymph node metastasis of oral carcinomas because of the high accumulation of active metabolites in metastatic and non-metastatic lymph nodes. PMID- 9861337 TI - Is radiation-induced degranulation of mast cells in salivary glands induced by substance P? AB - Although DNA is the critical target for the lethal effects of irradiation, the precise mechanisms by which irradiation causes damage in tissues and biological systems is not fully understood. In the present study, the number of mast cells and the expression of the neuropeptide substance P (SP) in salivary glands were examined 10 days after a regimen of irradiation. The irradiation was given as a single dose or 5 consecutive days with daily doses of 7 Gy up to a total dose of 35 Gy. In addition, the number of mast cells and the expression of SP were examined 2 and 24 h after a single dose of 7 Gy. Immunohistochemical staining for 5-hydroxytryptamine (5-HT) and staining with avidin peroxidase and toluidine blue were used to detect mast cells. At examination 2 and 24 h after irradiation treatment, no change in the number of mast cells and the pattern of SP expression was observed. Ten days after irradiation there was a remarkable reduction in the number of mast cells in all the three glands, but there was a marked increase in the number of nerve fibers showing SP-like immunoreactivity in the parenchyme. The results show that early time-dependent alterations in the density of mast cells occur in response to irradiation, and that these changes occur concomitantly with changes in the expression of SP. Since the peripheral nervous system is a main regulator of salivary gland function, it is tempting to speculate that the nervous system interacts with mast cells via SP in modulating irradiation provoked tissue responses in salivary glands. PMID- 9861338 TI - Smoking patterns and cancer of the oral cavity and pharynx: a case-control study in Uruguay. AB - A case-control study of cancer of the oral cavity and pharynx was conducted in Uruguay, between 1992 and 1996. 425 patients microscopically diagnosed with squamous cell carcinoma of the oral cavity and pharynx were frequently matched on age, residence, and urban/rural status with 427 hospitalised controls. The study was restricted to males. Smokers of black tobacco cigarettes were associated with an increased risk of 12.1 (95% confidence interval (CI) 7.6-19.4), when compared with non-smokers after fitting a model which included the matching variables, birthplace, education, and total alcohol consumption. Lifelong smokers of hand rolled cigarettes displayed an odds ratio (OR) of 8.7 (95% CI 5.6-13.4), compared with non-smokers. When smokers were excluded from the calculations, the OR for smokers of black tobacco cigarettes was 3.0 (95% CI 2.0-4.6), compared with smokers of blond tobacco cigarettes, after controlling for the same variables mentioned above, plus pack-years, years since stopping, and filter use. Hand rolling appears to be less important than smoking black tobacco in this study (OR 1.6, 95% CI 0.9-2.5). Thus, smoking black tobacco cigarettes appears to be an important habit in oral and pharyngeal carcinogenesis. PMID- 9861340 TI - Expression of cyclin dependent kinase inhibitor p21waf1/cip1 in premalignant and malignant oral lesions: relationship with p53 status. AB - p21waf1/cip1 protein, an inhibitor of cyclin dependent kinases, is a critical downstream target in the p53-specific pathway of growth control, and can also be induced by p53 independent pathways in relation to terminal differentiation. p21waf1 is also a putative tumour suppressor. Hence, we sought to determine whether this protein is abnormally expressed during betel- and tobacco-related oral oncogenesis. The aim was to determine whether a correlation exists between the expression profile of p21 and clinicopathological parameters of the patients, as well as with their p53 status. Immunohistochemical analysis showed that the expression of p21 protein in premalignant lesions was consistently elevated in the superficial, differentiated cells of the epithelium, while overexpression of the p53 tumour suppressor gene was observed in the basal proliferating layers of the epithelium. Our study demonstrated that p21 overexpression is associated with differentiation in proliferating dysplasias and squamous cell carcinomas (SCCs). The expression of p21 and p53 proteins was observed in 11/25 premalignant lesions. In 7 of these 11 cases, a heterogenous pattern of expression of p21 and p53 was observed. Four of these 11 premalignant and 30/51 malignant lesions showed concordant expression of both p21 and p53 proteins. The discordant p21 +/p53- phenotype was observed in 4/25 premalignant lesions and 5/51 oral SCCs. The p21-/p53+ phenotype was observed in 5/25 premalignant lesions and 7/51 oral SCCs. These results suggest that induction of p21 occurs by both p53 dependent and independent mechanisms during oral tumorigenesis. PMID- 9861339 TI - Survival analysis in a sample of oral cancer patients at a reference hospital in Rio de Janeiro, Brazil. AB - This article gives the results of a 5-year survival analysis of 371 oral cancer patients who received medical care at a cancer reference hospital in Rio de Janeiro between 1986 and 1987. Patient profiles based on selected risk factors for oral cancer and 5-year survival based on prognostically relevant variables are described. PMID- 9861341 TI - A comparison between the University of Washington Head and Neck Disease-Specific measure and the Medical Short Form 36, EORTC QOQ-C33 and EORTC Head and Neck 35. AB - The University of Washington Head and Neck Disease-Specific questionnaire (UW QOL) is potentially a suitable routine measure of outcome in head and neck cancer because it is quick and simple for patients to complete and is easy to process. The aim of this study was to compare the UW-QOL with three validated instruments in a group of patients having primary surgery for oral cancer. Between May and December 1995, 34 consecutive patients with previously untreated oral and oropharyngeal squamous cell carcinoma underwent primary surgery. Patients self completed the following questionnaires: UW-QOL, Medical Outcomes Short Form 36 (SF-36), the European Organisation for Research and Treatment of Cancer (EORTC) QOQ-C33 and the EORTC Head and Neck (H&N35). Questionnaires were distributed at four time intervals: pre-operatively, and at 3, 6 and 12 months. Patients were withdrawn if they developed recurrent disease. 29 patients agreed to participate in this prospective study and cross-sectional comparisons were made on baseline data and trends over time for longitudinal comparisons. This study emphasises the difficulties of one questionnaire to adequately evaluate quality of life. However, it would appear that the UW-QOL is a broad measure suitable for routine "low cost" assessment of disease-specific functional status. PMID- 9861342 TI - Expression of bcl-2 oncoprotein in Indian oral squamous cell carcinomas. AB - Thirty-nine oral squamous cell carcinomas were assessed for bcl-2 protein expression by immunostaining of tumour sections. Twenty-three per cent of these tumours showed strong nuclear staining for bcl-2 protein. Tumours of the cheek and tongue together accounted for 77% of overexpression of this protein. When bcl 2 expression was compared with p53 expression, they were found to be non overlapping. These results suggest that overexpression of either of these genes may substitute each other in the development of oral carcinomas of Indians. PMID- 9861343 TI - Laryngeal cancer in Lower Silesia: descriptive analysis of 501 cases. AB - This paper presents a descriptive analysis of 501 cases of laryngeal cancer. The patients were classified according to their age, sex, primary tumour localisation and stage. There were 448 males and 53 females, with a male-to-female ratio of 8.5:1. The peak incidence of cancer was detected in the 6th and 7th decades of age. The most common primary tumour localisation was the glottis. Three-hundred and-forty patients presented at advanced tumour stages (T3/T4). The proportion of patients presenting matastatic neck nodes was 29.3% and the incidence of metastatic lymph nodes increased with increasing T stage. Histopathological analysis revealed that 98% of tumours were squamous cell carcinomas. There were 125 well-differentiated, 235 moderately differentiated and 133 poorly differentiated carcinomas. Among other tumours, there was 1 adenoid squamous cell carcinoma, 1 giant cell carcinoma, 1 adenoma pleomorphicum, 1 adenoid cystic carcinoma (cylindroma), 1 haemangiopericytoma, 1 verrucous cell carcinoma, 1 lymphoepithelioma and 1 granular cell tumour. PMID- 9861344 TI - The use of pilocarpine hydrochloride to prevent xerostomia in a child treated with high dose radiotherapy for nasopharynx carcinoma. AB - A case is presented of a 10-year-old male with carcinoma of the nasopharynx with involvement of neck nodes, treated with high dose radiotherapy after four cycles of chemotherapy. Because of concern about causing xerostomia with its attendant problems, pilocarpine hydrochloride, 5 mg orally three times daily, was initiated at the onset of radiotherapy. Radiotherapy was well tolerated and at 7 months postradiotherapy, there was no evidence of residual or recurrent tumour and no xerostomia. Pilocarpine hydrochloride should be considered in children who are to receive high dose radiotherapy to the head and neck region which would include the parotid glands. PMID- 9861345 TI - A rare case of fibrosarcoma of the jaws in a 4-year-old male. AB - A case of fibrosarcoma in a 4-year-old male child is reported. Primary fibrosarcomas of bone in the head and neck region are rare. The histological appearance of the tumour is related to its grade of differentiation. It could present a high level of cellularity and the amount of collagen is variable. The accepted treatment is radical surgery; however, metastases could occur in the lungs. This article is presented to highlight the rarity of fibrosarcomas in the jaws of children. PMID- 9861346 TI - New superselective intra-arterial infusion via superficial temporal artery for cancer of the tongue and tumour tissue platinum concentration after carboplatin (CBDCA) infusion. AB - We developed a new technique of superselective intra-arterial chemotherapy for tongue cancer using a modified (1.35 mm) angiographic catheter. The catheter was confirmed to be inserted into the lingual artery by the new technique. We measured the platinum concentrations in resected tumour tissues after infusion of carboplatin (CBDCA) at 20 mg/m2 over 30 min from 30 min before tumour resection in 12 patients with cancer of the tongue (6 patients: superselective intra arterial infusion; 6 patients: conventional intra-arterial infusion). The mean platinum concentration in tumour tissue was 10.5 +/- 1.2 micrograms/g wet, which was more than twice higher than, and significantly different from, 4.3 +/- 3.8 micrograms/g wet by the conventional intra-arterial infusion method. This new superselective intra-arterial infusion method allows direct infusion of the anticancer agent into the artery supplying the tumour and is expected to become a new therapeutic modality for cancer of the tongue. PMID- 9861347 TI - Human papillomavirus, Epstein-Barr virus, human herpesvirus 8 and human cytomegalovirus involvement in salivary gland tumours. AB - Human papillomaviruses (HPV) are involved in the etiology of both benign and malignant epithelial lesions. The occurrence of HPV types 16 and 18 in gynecological squamous cell carcinomas is also well known. Of the herpesviruses, Epstein-Barr virus (EBV) is associated with, for example, undifferentiated nasopharyngeal carcinoma, endemic Burkitt's lymphoma and immunoblastic lymphoma, and human herpesvirus 8 (HHV-8) with Kaposi's sarcoma. As little is known about the etiological factors of salivary gland tumours, the presence of HPV, EBV, HHV 8 and human cytomegalovirus (CMV) in these tumours were examined. Fresh tissue samples obtained from 19 consecutive pleomorphic adenomas and 19 malignant salivary gland tumours were analyzed with polymerase chain reaction. Two samples showed EBV DNA positivity, a lymphoma of the parotid gland and a pleomorphic adenoma arising in the nasal cavity. HPV, HHV-8 and CMV DNA were not detected in any of the tumour samples. The results indicate that HPV, HHV-8 and CMV do not seem to have any role in the etiology of salivary gland neoplasms. PMID- 9861348 TI - pRb and p16 protein alterations in human oral tumorigenesis. AB - Cyclin dependent kinase inhibitor 2/multiple tumour suppressor gene 1 (CDKN2/MTS1) and retinoblastoma (Rb) tumour suppressor genes play important roles in the regulation of the cell cycle. The protein products of these genes p16INK4 (p16) and pRb, respectively, like p53 protein inhibit progression from G1 to S phase. p16 exerts its function through inhibition of CDK4-mediated phosphorylation of pRb. The pRb/p16 pathway is a critical target for molecular aberration at the G1-S checkpoint in a wide range of primary human tumours. The expression of p16 and pRb proteins was analyzed by immunohistochemistry in 35 cases of oral squamous cell carcinomas (SCCs), 22 cases of premalignant oral lesions and 30 normal oral tissues. Lack of pRb expression was observed in 23/35 (66%) oral SCCs and 14/22 (64%) premalignant lesions. Lack of p16 expression was observed in 22/35 (63%) oral SCCs and 13/22 (59%) premalignant lesions. Weak p16 and pRb immunoreactivities were observed in normal oral mucosal epithelium. The status of p16 and pRb was correlated with clinicopathological characteristics of the patients. Alteration in p16 expression showed significant correlation with tumour staging and progression (P = 0.024). Alteration in pRb/p16 expression correlated with heavy consumption of betel and tobacco. Our results suggest that alterations in the p16/pRb pathway are early events in oral tumorigenesis and may be involved in the development of betel- and tobacco-related oral malignancies. PMID- 9861349 TI - Differentiation of odontogenic keratocysts from other odontogenic cysts by the expression of bcl-2 immunoreactivity. AB - Odontogenic keratocysts (OKC) present an aggressive course with a marked tendency to recurrence. The epithelium of OKC is thought to have an intrinsic growth potential and has been shown to present a higher rate of proliferation as compared to other types of cyst. bcl-2 has a role in the extension of cell survival. The objective of the present study was to evaluate the bcl-2 protein expression in different odontogenic cysts. A total of 19 dentigerous cysts (DC), 20 radicular cysts (RC) and 14 OKC were used in the present study. DC and RC showed an almost complete negativity for bcl-2. OKC, on the other hand, presented in all cases a strong positivity in the cells of the basal layer, with, in most cases, more than 50% of the cells positive. This bcl-2 positivity of the basal layer of OKC could point to an abnormal control of the cell cycle. The bcl-2 protein overexpression could then produce an increase in the survival of the epithelial cells, and this increased lifespan could, in turn, lead to the peculiar aggressive growth pattern of OKC. Moreover the bcl-2 staining can be useful to differentiate OKC from other types of odontogenic cysts. PMID- 9861350 TI - Expression of proliferating cell nuclear antigen in ameloblastomas and odontogenic cysts. AB - The identification of the proliferative activity in tumours may be useful to predict the biological behaviour of different lesions. Proliferating cell nuclear antigen (PCNA) has been used for the evaluation of the proliferative ability of many lesions. In this study 22 ameloblastomas (4 follicular, 5 plexiform, 4 acanthomatous, 5 unicystic, 4 recurrent), 12 odontogenic keratocysts (OKC), 8 dentigerous cysts (DC), and 12 radicular cysts (RC) were analysed. PCNA+ cells were present in all cyst types but the OKC contained the highest number of PCNA+ cells. In OKC the location of PCNA+ cells was mainly suprabasal. In ameloblastoma PCNA+ cells were located mainly in the peripheral portion of the tumour islands. Statistical analysis showed that ameloblastoma had higher PCNA+ cell counts than OKC (P < 0.0001); OKC had higher values than DC and RC (P < 0.0001). Recurrent ameloblastoma presented higher PCNA+ cell counts than other types of ameloblastoma, while unicystic ameloblastoma showed lower values than acanthomatous, plexiform and follicular ameloblastomas (in this latter case the difference was not statistically significant). These data could help to explain the different biological behaviour of these lesions. PMID- 9861351 TI - Detection of HPV-16 genome in human oral cancers and potentially malignant lesions from India. AB - The presence of high risk human papilloma virus (HPV) 16 and 18 was examined in 100 oral cancer patients of Indian descent, 80 patients with potentially malignant oral lesions and corresponding clinically normal mucosa from 48 of these patients. Additionally, presence of HPV-33, -6 and -11 was also studied in 86 oral cancers, 50 potentially malignant oral lesions and 30 corresponding normal oral mucosa. All the patients with oral cancer and oral lesions, were long term tobacco-chewers, and a majority of the patients were in Advanced Stages III and IV. The DNA samples were amplified by polymerase chain reaction (PCR) using HPV L1 consensus primers. Typing of HPV was performed by Southern hybridization analysis of the PCR products using HPV-16, -18, -33, -6 and -11 type specific oligonucleotide probes. HPV-16 was detected in 15 out of 100 (15%) oral tumours, 27 out of 80 (34%) potentially malignant lesions and 15 out of 48 (31%) of the corresponding normal mucosa in the patients with oral lesions. HPV-18 was not detected in any of the oral cancers, oral lesions and normal mucosa. HPV-33 and the low-risk HPV-6 and -11 were also not detected in the oral cancers, oral lesions and corresponding normal mucosa. A significantly higher prevalence of HPV 16 was observed in oral lesions (27 out of 80, 34%) as compared to oral cancers (15 out of 100, 15%). The observed difference of 19% (95% confidence interval [CI]: 6%, 31%), between these two proportions was statistically significant at the 5% level of significance. Our data indicates that HPV-16 may play a direct role in a certain proportion of oral cancers; whereas in a subpopulation of oral cancers HPV-16 infection may be vital in the early events associated with development of potentially malignant oral lesions, and the presence of the virus not essential in the progression of the oral lesion to frank malignancy. PMID- 9861352 TI - Epidemiology of cancer of the lip in The Netherlands. AB - Descriptive epidemiological data of new cases of squamous cell carcinoma of the vermilion border of the lip in the Netherlands from 1989-94 inclusive are presented. Lip cancer represented 0.47 and 0.09% of all new malignancies in males and females, respectively. The lower lip was the most frequently affected site. The majority of the lip cancers were diagnosed in tumour stage I. The median age at diagnosis in males was 68 years, 5 years less than in females. The overall male-to-female ratio was 5.7. Age-adjusted incidence rates in males and females were 2.2 and 0.3 per 100,000 (ESR), respectively. The cumulative lifetime risk for developing lip cancer was 0.15 for males and 0.03 for females. Mortality/incidence ratios in males and females were 0.05 and 0.07, respectively. Differences in lip cancer incidence were observed between an urban and a rural area. There was a positive association between the occurrence of lip cancer and rural residence; rate ratios were 3.3 among males and 3.5 among females. PMID- 9861353 TI - Oral ulceration as a presenting sign of unknown sarcoidosis mimicking a tumour: report of 2 cases. AB - Oral lesions seem to be rather infrequent in sarcoidosis, and only 43 cases have been described in the English literature. Clinically most oral lesions appear as non-tender well-circumscribed swellings or submucosal nodules. Superficial ulceration is unusual. The authors present two cases in which the first presenting symptom of an unknown sarcoidosis was the presence of an oral ulcerated lesion with indurated margins that could easily have been misdiagnosed for a squamous cell carcinoma. It must be stressed that in all ulcerated lesions of the oral cavity, squamous cell carcinoma must always be considered in the differential diagnosis. PMID- 9861354 TI - Solitary fibrous tumour of the tongue. AB - Solitary fibrous tumour (SFT) is a neoplasm most often localised in the pleura and peritoneum. The tumour is composed of spindled fibroblastic cells arranged in a haphazard way. Recently SFT has been described in many locations. Only one case of oral SFT has been described in the cheek: this is the second case of an oral SFT located in the tongue. The differential diagnosis must be made from many soft tissue tumours. SFTs stain strongly, in almost all cases, for CD34. PMID- 9861355 TI - The International Cochrane Collaboration. PMID- 9861356 TI - No influence of socioeconomic factors on severe malarial anaemia, hyperparasitaemia or reinfection. AB - Malaria is responsible for nearly 500 million clinical cases per year, only a small proportion of whom will become severely ill. Socioeconomic risk factors may play a role in the development of severe malaria in African children and in their susceptibility to reinfection. In Gabon, 100 children suffering from severe malaria, defined as hyperparasitaemia and/or severe anaemia, were matched for sex, age and provenance to 100 children with mild malaria. Socioeconomic factors were assessed using a standard questionnaire and compared between the 2 groups. The children were followed-up and the time to first reinfection was recorded. No significant influence of socioeconomic factors could be detected on the severity of disease or the time to first reinfection. Socioeconomic factors are not major determinants of severe malarial anaemia and hyperparasitaemia in children in Gabon. PMID- 9861357 TI - Direct comparison of microscopy and polymerase chain reaction for the detection of Plasmodium sporozoites in salivary glands of mosquitoes. PMID- 9861358 TI - The use of morbidity questionnaires to identify communities with high prevalences of schistosome or geohelminth infections in Tanzania. AB - Parasitic infections were investigated in Morogoro Rural District, Tanzania, between October 1992 and June 1993. A total of 4589 schoolchildren (aged 7-17 years) from 30 primary schools was screened for infection with Ascaris lumbricoides, Trichuris trichiura, hookworms (3456 children only), Schistosoma mansoni and S. haematobium. The children were also asked about their recent experiences of the following: diarrhoea, abdominal pain, blood in stool, perception of suffering from schistosomiasis, and worm infection and examined for spleen and liver enlargement. Among schools, there were correlations between the prevalence of S. mansoni infection and bloody stools, spleen enlargement and liver enlargement, and between S. haematobium infection and the presence of blood in urine. To exclude ecological explanations for the correlations, logistic regression was used to estimate the adjusted odds ratio (OR) for each infection and each sign or symptom. No sign or symptom was significantly associated with any geohelminth infection. Reported blood in stool was significantly associated with S. mansoni infection (OR = 1.62, P = 0.045). Reported blood in urine was significantly associated with S. haematobium infection (OR = 7.71, P < 0.001), as was reported blood in stool (OR = 11.52, P < 0.001), indicating that presence of blood in either form of excreta was related to the local term for schistosomiasis. These results support the possibility of using reported blood in stool as a means of rapid assessment for identifying communities with a high prevalence of S. mansoni infection. PMID- 9861359 TI - The population biology and epidemiology of schistosome and geohelminth infections among schoolchildren in Tanzania. AB - A study of helminth infections was undertaken among 3244 schoolchildren from 28 schools in Morogoro Rural District, Tanzania. Schistosoma haematobium was the most common infection, followed by hookworms, Ascaris lumbricoides, S. mansoni, and Trichuris trichiura. Infection prevalence of each species varied among schools and age groups, but not between sexes. There was no relationship between the prevalences of different infections among schools, except for a strong negative correlation between the prevalence of hookworm and S. mansoni infections. Within each age group, there was little excess overlap in the distribution of each infection; thus the number of multiple infections was low whereas the number of individuals harbouring at least one infection was relatively high. More children than expected carried infections of A. lumbricoides and S. mansoni, and the clustering effect increased with age. Only 2 schools had high overall infection prevalences of both geohelminths and schistosomes. Logistic regression analysis of morbidity and parasitological data indicated that individuals with multiple species infections were not at increased risk of morbidity (on a multiplicative scale) compared to individuals with single species infections. This was attributed in part to the low egg counts observed for each parasite species. The results implied little interaction between schistosome and geohelminth infections in the region, both in parasitological terms and in the context of their combined effects on health. Implications for the feasibility and benefits of combined control of geohelminths and schistosomes are discussed. PMID- 9861360 TI - Prevalence and genotype of hepatitis C virus infection in pregnant women and blood donors in Ghana. AB - The seroprevalence of hepatitis C virus was evaluated in blood donors and antenatal clinic attenders in Kumasi, Ghana and seropositive subjects were tested for hepatitis C virus ribonucleic acid by the polymerase chain reaction (PCR). The overall seroprevalence among Ghanaians was 2.8% but there was a significantly higher prevalence in males (4.6%) than in females (1.0%). No risk factor for infection was identified by a questionnaire. Among those who showed evidence of active infection with a positive PCR, the most common genotype was type 2 but the subtype could not be specifically determined; these type 2 hepatitis C viruses may be indigenous to Africa. PMID- 9861361 TI - Increasing frequency of tuberculosis among staff in a South African district hospital: impact of the HIV epidemic on the supply side of health care. AB - To describe the changing frequency of tuberculosis among staff in a South African hospital, and to compare incidence in health workers with that in ancillary staff, the number and type of cases of tuberculosis among staff diagnosed between 1991 and 1996 were ascertained. The incidence rate of tuberculosis among health workers and ancillary staff was compared with the age-specific rate in the community (20-59 years old). In 1991-1992, 2 cases of tuberculosis were diagnosed among hospital staff; but in 1993-1996 there were 20 cases diagnosed (annualized incidence rates 138/100,000 and 690/100,000; P < 0.0001). Of 14 cases tested (64%), 12 (86%) were infected with human immunodeficiency virus (HIV). Most cases (82%) successfully completed treatment, but 4 died (18%). The incidence of tuberculosis amongst health workers (558/100,000 person-years of observation [PYO]) and ancillary staff (445/100,000 PYO) was not significantly different (P = 0.7), but it was lower than the incidence rate among 20-59 years old people in the community (1543/100,000). Tuberculosis has increased amongst hospital staff, secondary to the impact of HIV. The HIV epidemic is having a substantial impact on the health of hospital staff and interventions to counter this are urgently needed. PMID- 9861362 TI - Community-acquired septicaemia in southern Viet Nam: the importance of multidrug resistant Salmonella typhi. AB - In a prospective study conducted between mid 1993 and 1994, 437 adults and children were admitted with community-acquired septicaemia to an infectious diseases hospital in southern Viet Nam. Gram-negative aerobes accounted for 90% of isolates and were predominantly Salmonella typhi (67%), Sal. para-typhi A (3%), Escherichia coli (10%), and Klebsiella spp. (5%). Other Salmonella spp. (1%), Pseudomonas aeruginosa (1%), Neisseria meningitidis (0.5%) and Haemophilus influenzae (0.2%) were uncommon. Staphylococcus aureus (5.5%) and Streptococcus pneumoniae (2%) were the most common Gram-positive isolates. Patients with enteric fever were younger (median age 16 years, range 1-63) than the other patients (median age 43 years, range 1-88) (P < 0.001) and had a lower mortality rate (0.3% vs. 23%; relative risk 69.5, 95% confidence interval 9.5-507.8; P < 0.0001). Over 70% of the Sal. typhi isolated were multi-drug-resistant, and 4% were resistant to nalidixic acid. Multidrug-resistant Sal. typhi is a major cause of community-acquired septicaemia in Viet Nam. PMID- 9861363 TI - Malaria diagnosis by the polymerase chain reaction: a field study in south eastern Venezuela. AB - A polymerase chain reaction (PCR) method that amplifies genus- and species specific sequences present within the small subunit of ribosomal ribonucleic acid (ssRNA) genes of the human malaria parasites was used for the diagnosis of malaria in south-eastern Venezuela. One hundred blood samples were submitted to deoxyribonucleic acid extraction, PCR amplification and electrophoretic analysis of the PCR products, and the results were compared to those of routine microscopical diagnosis. The sensitivity of PCR for detection of Plasmodium vivax and P. falciparum malaria was 99% and 100%, respectively. However, 6 patients (6%) harboured parasites undetected by microscopy. The PCR assay detected a high proportion of mixed infections: 29% (17/59) of the infections microscopically diagnosed as P. vivax were shown to be mixed infections of P. vivax and P. falciparum. Forty per cent (7/17) of the individuals with a missed P. falciparum infection had received chloroquine in the previous 30 d. These results suggest that, in places where transmission of both P. vivax and P. falciparum occurs, PCR detection of malaria parasites can be a very useful complement to microscopical diagnosis in order to ascertain the true incidence of each species and for the follow-up of patients after specific treatment. PMID- 9861364 TI - Simplification of the miniature anion-exchange centrifugation technique for the parasitological diagnosis of human African trypanosomiasis. PMID- 9861365 TI - The use of whole blood absorbed on filter paper to detect Wuchereria bancrofti circulating antigen. AB - The Og4C3 enzyme-linked immunosorbent assay (ELISA) to detect circulating Wuchereria bancrofti antigen uses 50 microL of serum. In this study, a whole blood sample absorbed on filter paper was tested as a substitute for serum. Serum samples were obtained from 60 Sri Lankan subjects by venepuncture and finger prick blood samples from the same individuals were directly absorbed on filter paper. Og4C3 ELISAs using serum and filter paper blood were compared. Despite the fact that the estimated amount of serum available for the ELISA with filter paper blood was only one-fifth of that available when serum was used, the 2 ELISAs gave almost identical results. Of the 39 positive serum samples, 38 were detected using filter paper blood. Employing the ELISA using filter paper blood, 619 people in Matara, Sri Lanka, were examined for antigenaemia. The positivity rate was 22.5%, 3.1 times higher than the rate of microfilaraemia detected by examination of 60 microL blood films. PMID- 9861366 TI - High prevalence of hepatitis B viral DNA in cirrhotic patients without surface antigen. AB - The diagnosis of liver diseases induced by hepatitis B virus (HBV) is supported by the detection of HBV surface antigen (HBsAg) in serum. The present study aimed to investigate the presence of HBV deoxyribonucleic acid (DNA) in patients with liver cirrhosis using a polymerase chain reaction (PCR) based on primers derived from the pre-S1 and pre-core regions. HBsAg was detected in 10 of 48 patients (21%), total anti-hepatitis B core antigen (HBc) antibodies in 54%, anti hepatitis B e antigen (HBeAg) in 14.6%, anti-HBc immunoglobulin M in 8%, and anti HBs in 26%; none had detectable HBeAg. HBV DNA was detected in 73% of the cirrhotic patients. All cirrhotic patients with HBsAg also had HBV DNA; HBV DNA was detected in 64.5% of those without HBsAg. We conclude that the clearance of HBsAg does not necessarily indicate termination of viraemia in patients with liver cirrhosis and the detection of HBV DNA using a PCR based on primers from the pre-S1 and pre-core regions should be included in the diagnosis of HBV infection. PMID- 9861367 TI - The significance of guinea worm infection in the immunological diagnosis of onchocerciasis and bancroftian filariasis. AB - Infections with Dracunculus medinensis frequently occur in the same geographical area as infections with Onchocerca volvulus and Wuchereria bancrofti. This study analysed the significance of D. medinensis infections for the specificity and sensitivity of available tests for antibody-based diagnosis of onchocerciasis (using individual recombinant clones OV-10, OV-11 and OV-16, and the OV-7/OV 10/OV-16 tri-cocktail, in an enzyme-linked immunosorbent assay) and for circulating antigen-based diagnosis of bancroftian filariasis (using the TropBio and the ICT card tests). Some immunological cross-reactivity was observed with all tests. When using individual recombinant O.volvulus antigens, the highest assay indices were obtained for clone OV-10, and the lowest for clone OV-16. Testing the serum responses against the tri-cocktail of recombinant antigens did not notably improve the assay indices. Two of 40 serum samples from individuals with patent dracunculiasis gave a false positive response in the ICT test and one of these was also positive in the TropBio test. Possible implications of applying these diagnostic assays in areas endemic for dracunculiasis are discussed. PMID- 9861368 TI - Plasmodium ovale in a highly malaria endemic area of Senegal. AB - During 4 months, from June to September 1990, the population of Dielmo village, Senegal, an area of intense and perennial malaria transmission, was enrolled in a follow-up study including daily clinical surveillance and bi-weekly malaria parasitaemia monitoring. Thick blood film examinations indicated that 48.5% of children (49/101) and 32.4% of adults (34/105) were infected at least once by Plasmodium ovale during the study period; 148 distinct episodes of patent parasitaemia were observed, with estimated maximum durations of 3-115 d. The mean duration at first decreased significantly with age, from 11.4 d in children under 5 years old to 4.2 d in adults aged 40-59 years, but then increased in older adults to 7.0 d. In all age groups, most infections were asymptomatic. Only high parasitaemias were significantly associated with fever; 3 clinical malaria attacks due to P. ovale were seen during the study period. PMID- 9861369 TI - All four species of human malaria parasites form rosettes. PMID- 9861370 TI - Clinical case definitions for malaria: clinical malaria associated with very low parasite densities in African infants. AB - In areas endemic for Plasmodium falciparum, clinical malaria is believed to be less common in infants than in older children, but specific case definitions have rarely been determined for this age group. As malaria case definitions are known to be both age- and site-specific, assessment of the risk of disease in infancy requires the development of appropriate diagnostic criteria. In southern Ghana, 154 children were recruited at birth and monitored for fever and malaria infection until 2 years of age. Logistic regression was used to model fever risk as a continuous function of parasite density to determine case definitions for the diagnosis of clinical malaria, and to determine age- and season-specific estimates of the fraction of fevers attributable to malaria (AF); 2360 observations were made on 154 children. For fevers defined by a measured temperature > or = 37.5 degrees C, the estimated population AF was 44% (95% confidence interval 34-53). Estimates of AF varied with age and season. For infants, AF was 51% during the wet season and 22% during the dry season; for children over one year of age, AF was 89% during the wet season and 36% during the dry season. The estimated parasite density threshold for initiation of a febrile episode was 100 parasites per microL of blood in infants, compared with 3500 parasites per microL for children over one year of age. Using these case definitions, the incidence of clinical malaria was estimated at 0.09 cases per child-year at risk for children less than 6 months of age, 0.40 for children aged 6-11 months, and 0.69 for children aged 12-23 months. Of 66 cases of clinical malaria, only 3 were observed in children under 5 months of age. We concluded that, although most fevers in infants are not due to malaria, infant clinical malaria may occur at extremely low parasite densities. This may be indicative of a lack of anti-disease immunity in this age group. In southern Ghana, an infant with axillary temperature > or = 37.5 degrees C and parasitaemia > or = 100/microL should be considered to have clinical malaria. Nevertheless, the incidence of clinical malaria is very low in children under 6 months of age, confirming that they are significantly protected from clinical malaria compared to older children. PMID- 9861371 TI - Non-radioisotopic glucose turnover in children with falciparum malaria and enteric fever. AB - To determine whether glucose turnover is increased in acute falciparum malaria compared to enteric fever in children, steady-state 6,6-D2-glucose turnover was measured in 9 Malaysian children with uncomplicated malaria (6 males and 3 females; median age 10 years, body weight 22 kg) and in 12 with uncomplicated enteric fever (8 males and 4 females; median age 10 years, body weight 24 kg) in acute illness, after quinine (5 malaria patients) and in convalescence. Baseline plasma glucose concentrations in malaria and enteric fever were similar (all values are medians [ranges in brackets]) 5.6 [3.2-11.3] vs. 5.5 [4.2-8.0] mmol/L), as were serum insulin levels (5.6 [0.4-26.5] vs. 6.8 [1.1-22.5] milliunits/L; P > 0.4). Glucose turnover in the malaria patients was higher than in patients with enteric fever (6.27 [2.71-6.87] vs. 5.20 [4.50-6.08] mg/kg.min; P = 0.02) and in convalescence (4.74 [3.35-6.79] mg/kg.min; P = 0.05 vs. acute malaria study), and fell after quinine together with a rise in serum insulin (P = 0.03). Basal plasma lactate concentrations were higher in enteric fever than in malaria (3.4 [1.8-6.4] vs. 0.8 [0.3-3.8] mmol/L; P < 0.0001) and correlated inversely with glucose turnover in this group (rs = -0.60; n = 12; P = 0.02). These data suggest that glucose turnover is 20% greater in malaria than in enteric fever. This might reflect increased non-insulin-mediated glucose uptake in falciparum malaria and/or impaired gluconeogenesis in enteric fever, and may have implications for metabolic complications and their clinical management in both infections. PMID- 9861373 TI - Exfoliative skin manifestations in acute lymphatic filariasis. PMID- 9861372 TI - Trypanosoma brucei ssp. and T congolense: mixed human infection in Cote d'Ivoire. PMID- 9861374 TI - Serum procalcitonin concentrations in patients with pulmonary tuberculosis. PMID- 9861375 TI - Prognostic significance of clinical grading of patients envenomed by Bothrops lanceolatus in Martinique. Members of the Research Group on Snake Bite in Martinique. AB - The correlation between clinical grading of patients bitten by Bothrops lanceolatus and the subsequent development of their envenoming was examined. Severity of envenoming was graded using a 1-4 scale (minor to major). Patients were classified into 2 groups according to the time elapsed between bite and treatment with a specific purified equine F(ab')2 antivenom. The late/no treatment group (n = 33) was characterized by a systemic thrombotic complication rate of 14/33 (42.4%) leading to 4 deaths, which increased with the maximum severity assessed on the first day following the bite (P = 0.003). However, infarctions could develop in patients who presented initially with signs of moderate envenoming, normal blood clotting and low serum levels of venom antigens. No such complication of fatality occurred in the early (0.5-6 h) treatment group (n = 70). Multiple regression analysis showed that duration of stay in hospital in this group increased with the length of the snake (P = 0.017), venom antigenaemia (P = 0.016), initial grading (P < 0.001), and with the need for surgical debridement (n = 10/70, P < 0.001). Outcome was correlated with initial severity of envenoming. However, the only factor with a positive prognostic significance for the individual envenomed patient was the early infusion of specific antivenom, which led to 100% recovery in our series. PMID- 9861376 TI - A clinical and epidemiological study of Loxosceles spider envenoming in Santa Catarina, Brazil. AB - A clinical and epidemiological study of 267 cases of envenomation by Loxosceles spp. (loxoscelism), notified to Centro de Informacoes Toxicologicas de Florianopolis (Santa Catarina State, Brazil), was conducted between January 1985 and December 1995. Most of the incidents occurred along the coast of the mid southern region of the state, during the warmest months. L. laeta and L. intermedia were identified as the causative agents. Cutaneous loxoscelism was clinically diagnosed in 232 (86.9%) patients with local pain (86.5%), oedema (80.5%), hyperaemia (79.8%) and necrosis (56.9%). Cutaneous-visceral loxoscelism was detected in 35 patients (13.1%) with intravascular haemolysis, manifested by jaundice (68.6%), oliguria (45.7%), dark urine (28.6%), haemorrhage (25.7%), anuria (8.6%) and shock (2.9%), besides the cutaneous effects. Specific antivenom was given to 125 patients (46.8%) and only 8 (6.5%) had mild reactions. Acute renal failure was observed in 17 cases (6.4%); 4 patients (1.5%) died, all of whom were children under 14 years old. PMID- 9861377 TI - Geophagy, iron status and anaemia among pregnant women on the coast of Kenya. AB - In a cross sectional survey based in an antenatal clinic at Kilifi District Hospital, Coast Province, Kenya, 154 of 275 pregnant women (56%) reported eating soil regularly. Geophagous women had lower haemoglobin and serum ferritin concentrations than non-geophagous women (mean haemoglobin level 9.1 vs. 10.0 g/dL, P < 0.001; median ferritin level 4.5 vs. 9.0 micrograms/L, P < 0.001). In multiple linear regression analyses, geophagy was a significant predictor of haemoglobin (beta = -6.4, P = 0.01) and serum ferritin concentrations (beta = 6.6, P = 0.002), while controlling for gestational age and malaria and hookworm infection. Another 38 pregnant women, who reported eating soil regularly, participated in focus group discussions and were interviewed on geophagy. The most commonly eaten soil was from the walls of houses. The median estimated daily intake was 41.5 g (range 2.5-219.0 g). Twenty-seven of these women assisted in the collection of soil samples which were then analysed for their content of iron, zinc and aluminium after extraction with 0.1 M HC1. The average daily soil intake supplied the geophagous women with 4.3 mg of iron, corresponding to 14% of the recommended dietary allowance of iron for pregnant women. The study revealed a strong negative association between geophagy and both haemoglobin and ferritin status. At the same time it demonstrated the potential of soil as a source of dietary iron for geophagous women. These seemingly contradictory results might be due to other components in the soil interfering with iron uptake or metabolism. Alternatively, it may be that the geophagous women had extremely depleted iron stores before starting to eat soil. From these cross-sectional data, no inference about causality could be made. PMID- 9861379 TI - Chloroquine treatment for uncomplicated childhood malaria in an area with drug resistance: early treatment failure aggravates anaemia. AB - Childhood anaemia is a major public health problem in malaria holoendemic areas. We assessed the effects of antimalarial treatment in an area with drug-resistant falciparum malaria on haemoglobin levels in small children by applying the 1996 World Health Organization in vivo method for the evaluation of standard chloroquine treatment at the community level. In Fukayosi village, coastal Tanzania, 117 children aged 5-36 months with clinical malaria episodes were treated with chloroquine syrup (25 mg/kg). Early treatment failure (ETF) occurred in 20% and late treatment failure (LTF) in 22% of cases. Age > 1 year and malnutrition were protective factors against ETF. The evidence that chloroquine treatment could not prevent an exacerbation of anaemia was (i) the fact that the fall in haemoglobin level after 72 h was significantly greater in ETF than in children with LTF and an adequate clinical response, and (ii) the absence of any haematological improvement at follow-up in children receiving chloroquine alone, even in true treatment successes. In contrast, pyrimethamine/sulfadoxine administered to treatment failures improved the haemoglobin level significantly > 21 d after treatment started (mean difference 14 g/L, 95% confidence interval 2.1 27). We conclude that, when chloroquine treatment of childhood malaria is associated with a 20% ETF rate, the haemoglobin response is unsatisfactory and there is a need to change the recommended first-line treatment. PMID- 9861378 TI - Adolescent reproductive health: observations in a hospital setting. PMID- 9861380 TI - Plasmodium falciparum: modulation by phenobarbital of sensitivity to quinine, chloroquine and mefloquine in vitro. PMID- 9861381 TI - Early vomiting of mefloquine in children with malaria is not modified by the timing of antipyretic treatment. PMID- 9861382 TI - The relationship between capillary and venous concentrations of the antimalarial drug lumefantrine (benflumetol). PMID- 9861384 TI - Single dose of ivermectin to control mansonellosis in Trinidad: a four-years follow-up study. PMID- 9861383 TI - Treatment of human pulmonary paragonimiasis with triclabendazole: clinical tolerance and drug efficacy. AB - An open clinical trial to determine the efficacy and tolerability of postprandial doses of triclabendazole against Paragonimus mexicanus in 62 patients with pulmonary paragonimiasis from the Ecuadorian Amazon region was performed. Praziquantel was used as therapeutic control. Patients were allocated at random to the following 4 therapeutic regimens: triclabendazole, 5 mg/kg once daily for 3 d (16 patients), 10 mg/kg twice on one day (15 patients), and 10 mg/kg in a single dose (16 patients), and praziquantel, 25 mg/kg thrice daily for 3 d (15 patients). Clinical tolerance, based on the frequency and severity of adverse reactions, was superior in all 3 triclabendazole regimens to that of praziquantel. No alteration was observed in hepato-renal functions or haematological values. The clinical symptoms resolved at a comparable rate in all 4 treatment groups. A more rapid parasitological response to treatment, as determined by the reduction in the average number of parasite eggs found in sputum, was seen in patients treated with triclabendazole than with praziquantel. By day 90, 60 patients had no egg detected in their sputum; 2 patients, treated with a single dose of 10 mg/kg, had a few and were re-treated with triclabendazole (5 mg daily for 3 d). On day 365, none of the patients had eggs in their sputum. Triclabendazole can be recommended as an alternative drug of choice for the treatment of pulmonary paragonimiasis; it is as effective as praziquantel in clearing infections and better tolerated. PMID- 9861385 TI - F1 antigenaemia in bubonic plague patients, a marker of gravity and efficacy of therapy. PMID- 9861386 TI - Plasmodium ovale in South Africa. PMID- 9861387 TI - Application of molecular methods to the study of diseases prevalent in low income countries. PMID- 9861388 TI - Diagnosis of schistosomiasis japonica in Chinese schoolchildren by administration of a questionnaire. AB - This paper describes a rapid, simple, cost-effective questionnaire for screening school-aged children at risk for Asian schistosomiasis in China. Five hundred and thirty-two children, aged 8-14 years, were selected from 3 schools in an area moderately endemic for Schistosoma japonicum in Hunan province. The questionnaire, comprising 15 multiple-choice questions, was administered by teachers in order to collect both ethnographic and epidemiological data relevant to current S. japonicum infections. This was followed by Kato-Katz thick smear stool examinations, miracidium hatching tests, and soluble egg antigen-enzyme linked immunosorbent assays in order to validate the efficacy of the questionnaire approach. The results from a combination of all 3 procedures indicated that the overall schistosomiasis prevalence in the 3 schools was 29.9% (138/472). Six risk factors (episodes of diarrhoea, frequency of water contact, school grade attained, weakness, past history of S. japonicum infection(s), and whether a subject had been previously treated for schistosomiasis) in the questionnaire were determined by logistic regression to be highly statistically significant predictors of individual current infection. The sensitivity (93.7%), specificity (91.9%) and low cost (c. US$ 0.6/true positive case) associated with the 6 variables model make the questionnaire approach a very useful diagnostic tool for screening marshland and lake communities at high risk for schistosomiasis in China before selective treatment with praziquantel or diagnostic follow-up. An even simpler 3 variables 'yes/no' model was derived from the questionnaire and found to be nearly as good at predicting individual infection (sensitivity 86.2% and specificity exceeding 97.6%) and extremely simple to use. If validated in other ecological settings in China the questionnaire, modified or as presented here, could be adopted by the national schistosomiasis control programme. PMID- 9861389 TI - Spraying tick-infested houses with lambda-cyhalothrin reduces the incidence of tick-borne relapsing fever in children under five years old. AB - Tick-borne relapsing fever (TBRF) is a major cause of morbidity and mortality of children in central Tanzania. This study assessed the effect of house interior residual spraying with lambda-cyhalothrin on domestic tick infestation and the incidence of TBRF in children under 5 years old. Two neighbouring villages were studied: one was sprayed with insecticide, the other left untreated. Samples of earth from the floors of 50 houses before and after spraying, and 50 houses in the control village, were sieved and ticks counted. The incidence of TBRF was estimated by microscopy of thick blood films taken from all children under 5 years old presenting with fever to the village dispensary or health workers. After 2 cycles of spraying, no tick was found in the treated houses; in the controls, tick numbers remained high. In the treated village there were 29 cases of TBRF among the 960 children under 5 years old in the 4 months before spraying, and one in the 4 months after spraying. In the control village there were 10 and 4 cases in the same periods among the 510 children. There was a significant reduction of 0.09 in the odds ratio of infection after spraying (95% confidence interval 0.01-0.87). PMID- 9861390 TI - The health and nutritional status of schoolchildren in Africa: evidence from school-based health programmes in Ghana and Tanzania. The Partnership for Child Development. AB - Surveys of the health of schoolchildren in Tanga Region, Tanzania and Volta Region, Ghana are reported. Two age groups of both sexes were studied: 8-9 and 12 13 years old. Children themselves tend to have a poor perception of their health status. This is confirmed by biomedical surveys. Evidence was common of chronic ill-health due to undernutrition, anaemia, parasitic infections and micronutrient deficiencies. The older age groups of both sexes were significantly more stunted (height-for-age z score < 2 below National Center for Health Statistics reference values) than the younger groups, indicating that linear growth continues to falter throughout the school-age years. Anaemia was common: 38% of children in Ghana and 75% of children in Tanzania had a haemoglobin level < 120 g/L. Younger children were more likely to be anaemic than older children, but no significant difference between the sexes was observed. Helminth infections which cause blood loss (Schistosoma haematobium and hookworms) were common and only 37% of children in Ghana and 14% in Tanzania had no evidence of worm infection. In Ghana, 71% of children had a low urinary iodine concentration; in Tanzania 38%. The burden of ill-health suggests that school health programmes in these countries which deliver anthelmintics and micronutrient supplements have the potential to improve the health, growth and educational achievements of schoolchildren. PMID- 9861391 TI - Child sexual abuse in KwaZulu-Natal, South Africa. AB - This paper discusses child sexual abuse in a hospital serving a largely rural population in Eshowe, KwaZulu-Natal, South Africa during a period of 9 years and 2 months. Child sexual abuse was a significant problem. The incidence of sexually transmitted diseases at presentation was high (65.9%). The abuser, when identified, was more commonly an acquaintance or stranger than a family member. The shortage of resources in 'third world' countries means that medical personnel must take the lead in identifying and managing children with sexual abuse. Frequently they must assume additional roles, such as that of social worker, if management is to be successful. However, high patient case loads in these situations make this difficult and therefore child sexual abuse may not be adequately dealt with. A possible solution to this problem is the use of outside agencies to research the problem and the establishment of regional special units to manage child abuse. PMID- 9861392 TI - Risk factors for malaria: a microepidemiological study in a village in Sri Lanka. AB - Environmental and socioeconomic risk factors for malaria were studied in a village in Sri Lanka. Over a period of one year, all 49 households in the village were visited every alternate day to obtain information on malaria episodes. Information on risk factors was obtained through questionnaires and direct observations. Age below 17 years (relative risk [RR] = 1.66, 95% confidence interval [95% CI] 1.18-2.35), use of bed nets (RR = 0.16, 95% CI 0.05-0.45) and traditional fumigants (RR = 0.58, 95% CI 0.37-0.93) were independent predictors of malaria. People using anti-mosquito pyrethrum coils had a higher risk for malaria than people living in houses where they were not used (RR = 1.46, 95% CI 1.03-2.07). The build-up of Anopheles culicifacies populations before the start of the transmission season had taken place in a stream near the village. Living close to the stream was a risk factor for malaria early in the transmission season, although this did not reach statistical significance (comparing < 250 m with > 500 m, RR = 2.13, 95% CI 0.96-4.71). PMID- 9861393 TI - Similar feeding preferences of Anopheles gambiae and A. arabiensis in Senegal. AB - This study in Senegal compared the feeding preferences of Anopheles gambiae and A. arabiensis while controlling for equal accessibility to hosts located outdoors under bed net traps. All fed A. gambiae complex females were identified with the aid of the polymerase chain reaction and their blood meal sources were identified by enzyme-linked immunosorbent assay. A total of 605 anophelines, including 281 A. gambiae and 301 A. arabiensis, were captured, 32.2% in the human-baited traps and 67.8% in bovine-baited traps. 30.3% of A. gambiae fed in the former and 69.7% fed in the latter; the corresponding figures for A. arabiensis were 29.6% and 70.4%. Thus, when the hosts were located outdoors and made equally available, the feeding preferences of A. gambiae and A. arabiensis were similar (P = 0.81). These results suggest that biases existed in previous studies, most of which suggested that A. arabiensis was more zoophilic than A. gambiae. Alternatively, the feeding behaviour of these 2 species may differ in various parts of Africa. PMID- 9861394 TI - Anthroponotic cutaneous leishmaniasis in Kabul, Afghanistan: vertical distribution of cases in apartment blocks. AB - Owing to the civil war, the inhabitants of Kabul in Afghanistan are suffering a major epidemic of anthroponotic cutaneous leishmaniasis (ACL) caused by Leishmania tropica. Surveys conducted among children in 2 high-rise apartment blocks in the city revealed that the prevalence of active lesions was much lower on upper stories: 84% lower in one block (chi 2 = 7.13, d.f. = 1, P = 0.008) and 54% lower in the other (chi 2 = 6.17, d.f. = 1, P = 0.01). Similar trends were apparent with regard to scars from old lesions. These results suggest that in Kabul most transmission of ACL takes place in the home. In addition, the results imply that there must be limited vertical movement of the vector within apartment blocks. Together, these findings suggest that indoor spraying should be an effective means of control and that insecticidal applications could probably be restricted to lower stories. PMID- 9861395 TI - Dynamics of Leishmania chagasi infection in small mammals of the undisturbed and degraded tropical dry forests of northern Colombia. AB - The infection rate with Leishmania chagasi and the population dynamics of small mammals were studied in an undisturbed forest reserve (Coloso) and an area of highly degraded forest (San Andres de Sotavento [SAS]) in northern Colombia, both endemic for visceral leishmaniasis. Live trapping of mammals was done every month, and species, age, sex and reproductive status determined. L. chagasi was detected in samples of skin or spleen by the polymerase chain reaction, after extraction of deoxyribonucleic acid using specific primers (DB8/AJS3), and dot blood hybridization. Didelphis marsupialis was found to be infected in Coloso (3/21, 14.3%) and SAS (13/137, 9.5%); its relative abundance was higher in SAS (93/113, 82% of the captures). Although Proechimys canicollis was also found to be infected in Coloso (3/34, 8.8%) and SAS (2/4), its relative abundance was much lower (4%) in SAS than in Coloso (56% of 77 animals captured). Sciurus granatensis, Marmosa robinsoni, Heteromys anomalus, Zygodontomys brevicauda and Metachirus nudicaudatus were less common, and no L. chagasi infection was detected in them. PMID- 9861396 TI - Schistosomiasis in northern Mozambique. AB - A survey of 12 schools in Cabo Delgado, northern Mozambique, was performed to assess the extent of schistosomiasis infection. Questionnaires validated by parasitological examination of stools and urine were used. Very high prevalences of Schistosoma haematobium infection were found in all schools (77.5% was the lowest recorded), so the questionnaires did not distinguish 'high risk' schools. However, they clearly indicated a good knowledge of the infection by students and teachers. The disease was recognized by teachers to be a public health problem in the area. S. mansoni infection was rare. It had not been reported in previous surveys, but its vector Biomphalaria was discovered in the area. Revised mapping of the disease is advisable in view of probable changes after many years of conflict. PMID- 9861397 TI - Hepatitis B virus infection in immigrants to the southern Brazilian Amazon. AB - To identify epidemiological patterns and risk factors associated with hepatitis B virus (HBV) infection in the southern Brazilian Amazon, a survey was performed in the county of Terra Nova do Norte in the northern part of the state of Mato Grosso. The population consists mainly of immigrants from the southern part of Brazil, where HBV prevalence is low. A random sample (n = 783) of the population was interviewed and tested for HBV markers. The overall prevalence of HBV infection was 54.7%. There were 31 hepatitis B surface antigen (HBsAg) carriers (3.9%), with a low rate of HBV e antigen positivity, and none with anti-hepatitis D virus. Subtypes ayw3 and ayw2 of HBsAg were predominant, suggesting that the immigrants carried HBV from their original region to the Amazon. Clustering of HBV infection within families was found. The association between HBV markers and having lived in a gold-miners' camp, even after adjusting for confounders, indicated that the gold-miners may play a role in HBV spread in areas of the Amazon where gold prospecting occurs. PMID- 9861398 TI - Evaluation of two tests based on the detection of histidine rich protein 2 for the diagnosis of imported Plasmodium falciparum malaria. AB - The ParaSight-F dipstick test (Becton Dickinson, USA) and the ICT Malaria Pf test (ICT, Australia) both detect histidine rich protein 2 (HRP-2), a water-soluble antigen expressed by Plasmodium falciparum trophozoites. The present study compared the diagnostic performance of both tests in persons returning to Belgium from countries endemic for malaria. During a period of 18 months both tests were performed on all patients returning from the tropics with a positive malaria blood film. Patients with fever without an obvious cause were used as controls. For the ParaSight-F test, considering P. falciparum trophozoites only, sensitivity was 95% and specificity 90%. Considering trophozoites of all species of Plasmodium, sensitivity was 71% and specificity 87%. Finally, considering patients with clinical malaria, the sensitivity of the test was 72% and specificity 87%. For the ICT Malaria Pf test, sensitivity was 95% and specificity 89% for P. falciparum trophozoites only, 71% and 86% for trophozoites of all species, and 72% and 87% for clinical malaria. Both tests gave highly comparable results. However, antigen detection assays cannot replace conventional microscopy in diagnosing imported malaria. Thick blood film examination is more sensitive and more specific, it allows estimation of parasitaemia and distinction between parasite growth stages, and it covers all species. Moreover, with treated patients the use of antigen tests might lead to problems in determining the efficacy of therapy. PMID- 9861399 TI - Parasitological diagnosis of human African trypanosomiasis: a comparison of the OBC and miniature anion-exchange centrifugation techniques. PMID- 9861400 TI - Diagnosis of Wuchereria bancrofti infection by the polymerase chain reaction using urine and day blood samples from amicrofilaraemic patients. AB - A sensitive and specific polymerase chain reaction (PCR) based on a highly repeated deoxyribonucleic acid (DNA) sequence (188 bp; SspI repeat) was tested for the detection of Wuchereria bancrofti DNA in blood and urine samples collected during the day from individuals in Coque, Recife, Brazil, an endemic area for W. bancrofti. All microfilaraemic individuals were also positive by PCR, irrespective of the samples used. The PCR system was capable of detecting W. bancrofti DNA in amicrofilaraemic individuals: c. 93% were positive by PCR when day blood samples were used and 59.7% when urine samples collected at 07:00 were used. Thus, nocturnally periodic W. bancrofti infection can be detected in blood samples collected during the day, which is convenient for large-scale screening. In addition, non-invasive urine collection provided suitable samples for PCR, which is clearly advantageous for preliminary mass diagnosis. PMID- 9861402 TI - The correlation between microscopical examination and erythrocyte band 3 (AE1) gene deletion in South-east Asian ovalocytosis. AB - South-east Asian ovalocytosis status was determined by microscopical examination of peripheral blood samples collected from 137 individuals in Papua New Guinea. The examination was performed separately by 2 microscopists, one of whom was very experienced in examining peripheral blood films for the diagnosis of south-east Asian ovalycytosis and the other was recently trained. The samples were also analysed by polymerase chain reaction (PCR) to determine ovalocytosis status by demonstrating a 27 base pair deletion in erythrocyte band 3 protein of the affected individuals. The microscopists were unaware of each other's results and of those obtained by PCR. Generally, there was very good agreement between the results obtained by both microscopists and the PCR. Although there was considerable inter-observer variation in the final ovalocyte count between the 2 microscopists, this did not affect their ability to discriminate between ovalocytic and normocytic individuals. Taking the PCR results as the standard, for the first, more experienced observer, the most efficient ovalocyte count cut off point was around 50%. At this ovalocyte count the sensitivity and specificity of microscopical examination were 93.6% and 92.2%, and the positive and negative predictive values 86.3% and 96.5%, respectively. The second microscopist generally underscored the ovalocyte counts and his most efficient cut-off point was 20%, with sensitivity and specificity of 85.1% and 93.3% and positive and negative predictive values of 87.0% and 92.3%, respectively. PMID- 9861401 TI - Detecting undetected HIV-1 variants in African children using degenerate polymerase chain reaction and sequence analyses. PMID- 9861403 TI - Clinical predictors of malaria in Gambian children with fever or a history of fever. AB - Diagnosis of malaria in children is difficult without laboratory support because the symptoms and signs of malaria overlap with those of other febrile illnesses such as pneumonia. Nevertheless, in many parts of Africa diagnosis of malaria must be made without laboratory investigation. Therefore, a scoring system has been developed to assist peripheral health care workers in making this diagnosis. Four hundred and seven Gambian children aged 6 months to 9 years who presented to a rural clinic with fever or a recent history of fever were investigated. A diagnosis of malaria was made in 159 children who had a fever of 38 degrees C or more and malaria parasitaemia of 5000 parasites/microL or more. Symptoms and signs in children with malaria were compared with those in children with other febrile illnesses to identify features which predicted malaria. Symptoms and signs were incorporated into various logistic regression models to test which were best independent predictors of malaria and these regression models were used to construct simple scoring systems which predicted malaria. A nine terms model predicted clinical malaria with a sensitivity of 89% and a specificity of 61%, values comparable to those obtained by an experienced paediatrician without laboratory support. The ability of peripheral health care workers to diagnose malaria using this approach is now being investigated in a prospective study. PMID- 9861404 TI - Abnormal respiratory patterns in childhood cerebral malaria. AB - Of 295 children with cerebral malaria, 117 (40%) had an abnormal respiratory pattern; 15 children exhibited more than one pattern during their clinical course. Four distinct patterns were seen. (i) Deep breathing (80 children); this was associated with severe metabolic acidosis, and resolved following treatment with intravenous fluids and/or blood. (ii) Hypoventilation with nystagmus and salivation (18 children); simultaneous electroencephalographic recording revealed continuous electrical seizure activity, demonstrating that these children were in subtle status epilepticus; anticonvulsant treatment resulted in return to normal of blood gases and recovery of consciousness. (iii) Hyperventilation with extensor posturing (20 children), which was associated with varying degrees of intracranial hypertension. (iv) Periodic respiration (14 children); all had clinical features suggestive of transtentorial herniation, and died following a respiratory arrest. Abnormal respiratory patterns can alert the clinician to complications of cerebral malaria that require treatment. Recognition of these patterns and rapid initiation of appropriate supportive therapy may help to reduce the high mortality rate of this disease. PMID- 9861405 TI - Association of the haptoglobin phenotype (1-1) with falciparum malaria in Sudan. AB - The haptoglobin phenotypes of Sudanese patients with complicated and uncomplicated falciparum malaria, and those of uninfected randomly selected individuals, were determined by electrophoresis of sera on polyacrylamide gels followed by benzidine staining of the gels. Among 273 malaria patients, the proportions with haptoglobin phenotypes (1-1), (2-1) and (2-2) were 60.8%, 29.7% and 9.5%, respectively, and in 72 cerebral malaria patients the proportions were 63.9%, 29.2%, and 6.9%. The distribution among 208 control individuals was 26.0%, 55.8% and 18.3%, respectively. The difference between patients and controls was highly significant (P < 0.001). The distribution of the different haptoglobin phenotypes among the randomly selected group of 208 Sudanese individuals was comparable to that in many other populations. The results suggests that the haptoglobin phenotype (1-1) is associated with susceptibility to falciparum malaria and the development of severe complications; alternatively, the other phenotypes may confer resistance. PMID- 9861406 TI - Absence of an association between intercellular adhesion molecule 1, complement receptor 1 and interleukin 1 receptor antagonist gene polymorphisms and severe malaria in a West African population. AB - Many genes have been shown to be involved in host susceptibility to the severe forms of Plasmodium falciparum malaria but it is likely that a large number of malaria-susceptibility genes remain to be determined. We conducted a large case control study of children with the severe forms of this disease-cerebral malaria and severe malarial anaemia--to attempt to identify these genes. Over 1200 children in The Gambia were typed for polymorphisms of the intercellular adhesion molecule 1 (ICAM-1), complement receptor 1 (CR-1) and interleukin 1 receptor antagonist (IL-IRA) genes. None of the polymorphisms typed was significantly associated with severe disease. These data differed significantly from the results of a previous study (Chi 2 = 8.81; P = 0.003) in which the ICAM-1 gene polymorphism was shown to be significantly associated with cerebral malaria in a case-control study of 547 subjects in Kenya. This suggests that there may be heterogeneity in genetic susceptibility to this condition between these 2 African populations. PMID- 9861408 TI - Serum thrombomodulin levels in patients with typhoid fever. AB - Elevated plasma or serum levels of thrombomodulin (TM) have been reported in some diseases. However, to our knowledge, plasma and serum levels of TM have not been investigated in febrile patients with typhoid fever. Serum TM and creatinine levels were determined in 7 male Japanese febrile patients with typhoid fever and 6 male Japanese healthy controls. The serum TM values of the patients and controls, in arbitrary units/mL, were 5.04 (SD = 1.69) and 2.93 (SD = 0.74), respectively (P < 0.025). The serum creatinine levels of the patients and controls were 0.93 (SD = 0.16) mg/dL and 0.77 (SD = 0.10) mg/dL, respectively, and the lack of any significant difference between them suggests that delayed clearance from the kidneys was not the predominant explanation for the elevated serum TM levels. Although the numbers of patients and controls were small and more studies are needed, we conclude that elevated serum TM levels during the course of a febrile infection with Salmonella serovar typhi may reflect the disease activity. PMID- 9861407 TI - Omentoplasty in the management of filarial lymphoedema. AB - A trial of omentoplasty was carried out on 20 patients with unilateral filarial lymphoedema to assess its role in the reduction of oedema volume after failed lymphonodo-venous shunt (LNVS) or as a primary procedure. Omentoplasty was done through a midline laparotomy. The omentum was mobilized from the colon, preserving both gastroepiploic vessels, and transferred to the thigh either through the lower end of the laparotomy incision or through a separate stab and placed subcutaneously in the upper third of the thigh. There was no operative mortality. Morbidity was mainly incisional hernia and superficial wound infection. Fourteen of 18 patients had more than 25% reduction in oedema volume during the immediate postoperative period, and 5 of the 18 had more than 50% reduction. However, there was a gradual loss of response with time. Age, gender, grade of lymphoedema, duration, and previous surgery did not influence the outcome. The incidence of incisional hernia could be reduced by transferring the omentum through the midline. There was a statistically significant reduction in postoperative adenolymphangitis attacks whether or not the oedema volume was reduced. In some patients the oedema was reduced sufficiently to permit subcutaneous excision of the lymphoedematous tissue. There appears to be a definite but limited role for omentoplasty in patients who have failed LNVS. PMID- 9861409 TI - Renal involvement in Russell's viper bite patients without disseminated intravascular coagulation. AB - We measured urinary beta-N-acetylglucosaminidase (NAG: EC 3.2.1.30), blood urea, serum creatinine, albuminuria index and creatinine clearance in 16 patients bitten by Russell's viper (Daboia russelii siamensis) without disseminated intravascular coagulation (DIC) at the time of admission and then once daily for 5 d. Urinary NAG activity and the albuminuria index showed considerable variation whereas the other standard indicators of renal function revealed no abnormality. It is concluded that structural damage to the kidney occurred in these victims in the absence of DIC, indicating a direct toxic effect of Russell's viper venom on the kidney. PMID- 9861410 TI - Loading dose of quinine in African children with cerebral malaria. AB - The majority of deaths from cerebral malaria occur within 48 h after admission to hospital. Because of the possibility of inadequate treatment within this period, the use of a loading dose of quinine has been proposed. We reviewed clinical and laboratory data for 113 children with cerebral malaria, who were treated with intravenous quinine, 10 mg/kg every 8 h, at Macha Mission Hospital in rural Zambia. In 1990-1991, 39 children were not given a loading dose of quinine while, in 1992-1993, 74 children received a loading dose of 20 mg/kg. Elevated serum iron levels, as reflected in transferrin saturation, were strongly associated with higher mortality. A loading dose of quinine was associated with faster recovery from coma and enhanced clearance of parasitaemia and fever. The loading dose was also associated with trends to lower mortality and higher haemoglobin levels, but these differences were not statistically significant. PMID- 9861411 TI - Pharmacology and toxicology of artelinic acid: preclinical investigations on pharmacokinetics, metabolism, protein and red blood cell binding, and acute and anorectic toxicities. AB - The pharmacokinetics, metabolism, protein binding, red blood cell (RBC) binding, stability in vitro, and acute and anorectic toxicity of artelinic acid (ARTL) were investigated in various animal species and human blood samples. Absorption and distribution following 10 mg/kg intramuscular or oral administration in dogs and rats were very rapid with t1/2 0.12-0.54; there were also a high AUC (11,262 ng/h/mL) and Vss (9.5 L/kg), low CL (15 mL/min/kg) and long elimination time (t1/2 = 2.6 h), compared with rat data. Oral bioavailability of ARTL was 79.7% in dogs and 30.1% in rats. The conversion of ARTL to dihydroartemisinin (DART) in dogs (0.1-0.5% of total dose) after 3 routes of administration (intravenous, intramuscular and oral) was 10-fold lower than that in rats. In rats dosed with [14C]ARTL, unchanged ARTL accounted for less than 13% of the total radioactivity after all 3 administration routes, suggesting that ARTL was extensively biotransformed. The half-lives of total radioactivity (21-49 h) in urine were much longer than that of unchanged ARTL in plasma (1.4-3.7 h), indicating that some long-lasting metabolites of ARTL were formed in rats. The mass balance data showed that 77-83% of total radioactivity was recovered in urine and faeces. High binding capacity (79-95%) and low binding affinity (1.1-9.3 x 10-7 M) of ARTL were measured in rat, rabbit, dog, monkey and human plasma. The RBC/plasma ratios of [14C]ARTL were 0.35 and 0.44 for dog and human plasma, respectively. ARTL was much more stable than artesunic acid (ARTS) in rat and dog plasma, and both ARTL and ARTS were more stable in dog plasma than in rat plasma in vitro. The 50% lethal dose (LD50) of ARTL in rats was about 535 mg/kg. Multiple intramuscular dosing for 7 d of 50 mg/kg/d of ARTL caused mild anorectic toxicity compared to ARTS in rats. In contrast to 4 other artemisinin derivatives, ARTL seems to be a good antimalarial candidate as it has the highest plasma concentration, the highest binding capacities in RBC, the highest oral bioavailability, the longest elimination half-life, the lowest metabolism rate and the lowest toxicity at equivalent dose levels. PMID- 9861412 TI - A 5 days primaquine regimen as anti-relapse therapy for Plasmodium vivax. PMID- 9861413 TI - Absence of ivermectin-associated excess deaths. PMID- 9861415 TI - Improved treatment of syphilis among pregnant women through on-site testing: an intervention study in rural South Africa. PMID- 9861414 TI - Treatment outcome of an unselected cohort of tuberculosis patients in relation to human immunodeficiency virus serostatus in Zomba Hospital, Malawi. AB - There is little information about treatment outcome in patients with smear negative pulmonary tuberculosis (PTB) or extrapulmonary tuberculosis (EPTB) treated under routine programme conditions in subsaharan Africa. A prospective study was carried out to determine treatment outcome in an unselected cohort of TB patients admitted to Zomba General Hospital, Malawi. Eight hundred and twenty seven adult TB patients (451 men and 376 women) were registered between 1 July and 31 December 1995. Standardized treatment outcomes of treatment completion, death, default, and transfer to another district were assessed in relation to type of TB, human immunodeficiency virus (HIV) serostatus, age and gender. Two hundred and fifty-four patients (31%) died by the end of treatment, half of the deaths occurring in the first month. Death rates were 19% among 386 patients with smear-positive PTB, 46% among 211 patients with smear-negative PTB, and 37% among 230 patients with EPTB; 77% of the patients were HIV seropositive. Among new patients, HIV-positive patients had higher death rates than HIV-negative patients (hazard ratio [HR] 2.5; 95% confidence interval [95% CI] 1.6-3.8). Smear-negative patients had the highest death rates (HR 3.9; 95% CI 2.7-5.5 compared to smear positive patients), followed by EPTB patients (HR 2.6, 95% CI 1.8-3.7 compared to smear-positive patients). Death rates increased with age but were similar in men and women. Adult patients in Malawi with smear-negative PTB and EPTB have low treatment completion and high death rates, related to high levels of HIV infection. National TB control programmes in areas of high HIV prevalence should no longer ignore treatment outcomes in patients with smear-negative PTB or EPTB. PMID- 9861416 TI - Equine antirabies serum treatment during an epizootic outbreak in the city of Ribeirao Preto, Brazil. PMID- 9861417 TI - Serum circulating egg antigen levels in two areas endemic for Schistosoma mansoni. AB - A monoclonal antibody-based enzyme-linked immunosorbent assay detecting Schistosoma mansoni circulating soluble egg antigen (CSEA) was applied in epidemiological studies. The serum CSEA levels were determined for 2 populations with a high prevalence (> 95%) and high intensity of infection as determined by faecal egg counts. In one population (Maniema, Zaire) transmission had been occurring for several decades, while in the other population (Ndombo, Senegal) transmission had started only recently. CSEA could be detected in 88% and 70% of the serum samples from Maniema and Ndombo, respectively. The sensitivity of the CSEA assay increased with rising egg count. The age-related CSEA profiles of the Maniema population followed a pattern similar to that of egg counts and of the adult worm antigen CAA (circulating anodic antigen). However, the recently infected Ndombo population showed a clearly different profile: while the CSEA prevalence reached a peak in children and adolescents, the mean CSEA levels did not vary significantly in the different age groups. CSEA levels were significantly lower in Ndombo than in Maniema. As egg antigens in serum are thought to be in part, or even primarily, derived from eggs in the tissues, these findings indicate a relatively smaller tissue egg load in Ndombo than in Maniema. PMID- 9861418 TI - Leishmania major MON-74 as a causative agent of cutaneous leishmaniasis in Burkina Faso. PMID- 9861419 TI - First identification of a species of Leishmania causing visceral leishmaniasis in the Sultanate of Oman, in a patient with AIDS. PMID- 9861420 TI - Cultivation of Plasmodium falciparum in vitro using microbiological gas sachets that generate microaerophilic conditions. PMID- 9861421 TI - Assessment of suitability for psychotherapy. I. Introduction and the assessment process. AB - Part I of this paper reviews the value of detailed psychotherapy-suitability assessments using criteria independent of illness severity. False-positive and false-negative determinations of suitability are minimized along with the associated problems. These assessments must be done by psychotherapists, and they typically take several sessions. An assessment process based on three components- diagnosis and the traditional history, creation of a model of the patient's psychopathology, and the use suitability criteria (reviewed in detail in part II) -is described. The value of diagnosis is discussed using the two most common ones of depression and personality disorder, along with the implications for the prescription of psychotherapy. PMID- 9861422 TI - Psychotherapy with cancer patients. AB - Cancer is associated with significant psychosocial morbidity. Although psychodynamic psychotherapy is a valuable intervention for some cancer patients, this modality has been underutilized because psychotherapy with medically ill patients has been linked historically to work with patients who have psychosomatic illnesses. Psychotherapy with patients who have cancer has unique features, such as the prominence of illness-related issues during the initial phase of treatment, the mixture of supportive and interpretive therapy, focused goals, and special issues pertaining to the transference/countertransference field. Understanding of those special issues will improve psychiatrists' ability to use long-term psychotherapy as an effective psychosocial intervention for patients with cancer. PMID- 9861423 TI - Origins of homophobia in males. Psychosexual vulnerabilities and defense development. AB - PURPOSE: To better understand the origins of homophobia among males. METHODS: Literature review and clinical illustration. RESULTS: Data suggest that there is a range of homophobic attitudes. CONCLUSIONS: We illustrate how homophobic attitudes can be associated with a hierarchy of defensive styles. We propose that these defensive styles are used to manage a range of psychosexual developmental anxieties in boys and men. PMID- 9861424 TI - Dynamic change in crisis intervention. AB - The author describes his work integrating psychodynamic theory and therapeutic methods in a crisis center. Crises can be seen as situational mediators that place an individual's typical defenses and resistances in question. This state of crisis can allow for dynamic change, beyond the relief of symptoms typically expected in crisis work. PMID- 9861425 TI - Weakenings and repairs in supervisory alliances. A multiple-case study. AB - The "weakening-repair" process in the working alliances of 10 supervisory dyads was examined. Trainees' level of professional development affected both the type of weakening event and supervisors' repair behaviors. Other important determinants of this process were the power differential between supervisor and trainee, as well as the evaluative function of the supervisor. PMID- 9861426 TI - Attitudes toward psychotherapists, psychologists, psychiatrists, and psychoanalysts. A meta-content analysis of 60 studies published between 1948 and 1995. AB - Sixty studies, published between 1948 and 1995, on attitudes toward mental health professionals, particularly psychotherapists, were systematically content analyzed. Descriptive results concerning the topics status, expertise, personality attributes, physical appearance, sex/gender, psychotherapeutic settings, and interactions between therapists and clients are presented. Public image problems of psychotherapists, research deficits, methodological problems, and sex/gender issues are critically discussed. PMID- 9861427 TI - Boundary transgressions in the psychotherapeutic framework: who is the injured party? AB - The authors emphasize the therapeutic value of establishing and maintaining boundaries between clinician and patient. No universal definition of a therapeutic boundary exists, and this ambiguous term spans a diverse spectrum from benign crossings to harmful violations. Understanding of the traumatic nature of faulty boundaries in psychotherapy has derived largely from the study of severe sexual transgressions by therapists. In contrast, this paper highlights the importance of recognizing a wider and more subtle variety of boundary crossings, especially those by patients, which can compromise the integrity of treatment and traumatize the vulnerable therapist. This is exemplified here in the case study of an incest survivor and a novice clinician. PMID- 9861428 TI - Psychotherapy in Russia. Historical backgrounds and current practice. AB - During recent years, the former Soviet states have witnessed enormous social and cultural changes, which have also greatly influenced the field of mental health, including psychotherapy. In this article, the historical backgrounds of Russian psychotherapy and its current practice are described. Psychotherapy in Russia and in Western countries share common roots, but have developed into different directions during the 70 years of Soviet regime. In more recent years, they have begun to slowly converge again. In the West, a trend away from insight-oriented, nondirective psychotherapy is taking place in favor of more directive approaches, aimed at changing overt behavior. In contrast, there is a tendency for therapies in Russian-speaking countries to become gradually less directive and authoritarian. In these countries there is an increasing interest in psychodynamic, insight-oriented therapies. PMID- 9861429 TI - The widening intellectual scope of psychoanalysis. PMID- 9861430 TI - [Current state of the art and perspectives on hemoglobin solutions]. PMID- 9861431 TI - [Anesthesia simulation--consequences of technological development]. PMID- 9861432 TI - [Hemoglobin solutions: volume replacement or oxygen therapy?]. AB - The development of haemoglobin solutions has progressed significantly in the last 15 years because of a perceived short fall in allogeneic blood within the next decades and increased concern about transmitted infectious diseases. Animal studies have shown that modern highly purified and chemically modified haemoglobin preparations are free of toxic side effects, provide adequate volume replacement and have vasoconstrictive effects that enhance systemic vascular resistance and mean arterial pressures after haemorrhage and in models of nearly complete blood replacement. Microcirculatory effects of haemoglobin-based oxygen carriers are dependent on the respective organ and species in which they are applied and on their degree of purification and chemical modification. Because of different physico-chemical properties in comparison with red cells, haemoglobin solutions provide sufficient tissue oxygenation in areas with critically restricted perfusion even when applied in small doses. First studies in volunteers and patients showed efficacy and tolerability of different newly developed haemoglobin solutions during acute normovolaemic haemodilution and in perioperative blood replacement. However, only little information exists to date in terms of metabolism of haemoglobin preparations and their potential immunogenicity and immunosuppressive side effects. Technical problems with the clinical use of haemoglobin solutions arise because of interference of plasma haemoglobin with routine laboratory tests and oximetry. Future indications for haemoglobin solutions as an oxygen therapeutic allow for application of small doses of such preparations and may help to avoid major technical problems. More clinical studies have to be undertaken to confirm the effectivity and safety of the different haemoglobin solutions and to find out the optimal indications beyond acute preclinical and perioperative blood replacement. PMID- 9861433 TI - [Continuous improvement in anesthesiological quality documentation]. AB - OBJECTIVE: The analysis of result variation in quality benchmarking projects in anaesthesia showed that ASA classification was often the most relevant parameter in distinction of risk groups. Thus the parallel description of the risk level of a patient both with the ASA classification and with particular risk parameters was examined critically. The hypothesis was tested that the documentation of both parameter groups in the running quality benchmarking projects does not lead to relevant information gain. As a pragmatic consequence we see the possibility to reduce the core dataset with significant reduction of the documentation workload. METHODS: With machine readable protocols or online computer documentation nearly all anaesthesias in hospitals in Hamburg were documented with the DGAI core data set and transferred to the project office of EQS Hamburg since 1992. We compared the predictory power of single and combined risk assessments for the incidence of particular AVBs (grade 3 to 5) in elective anaesthesias with that of ASA classification. RESULTS: In 257,878 elective anaesthesias AVBs were documented in 14.5% of cases. Besides one exception no superior prediction power for AVB incidence could be demonstrated for any special risk assessment as compared with the ASA-classification. This is also true for the AVBs decompensated cardiac insufficiency, myocardial infarction, pulmonary embolism and cardiac arrest which are associated with high lethality. CONCLUSION: We assume that the documentation of risk factors in the core data set as predictors can be abandoned without major loss of information. This would be a first step towards reduction of the amount of data in quality assurance to get a core of especially meaningful parameters. It would lead to an increase in acceptance of the method and thus to an increase in the validity of its results. PMID- 9861434 TI - [Tooth injury during intubation--diagnostic and therapeutic aspects]. AB - This study analysis 16 cases of dental injuries occurring during endotracheal intubation. The overall rate was 1 out of 6000 cases of intubation. As expected, the upper jaw teeth are most often involved. Partial dislocations were the most common lesion, together with dental fractures. A well-documented dental evaluation before delivery of anesthetics and appropriate precautions and protective devices during intubation will prevent most dental trauma related to endotracheal intubation. Also, early use of dental and risk management services often will ensure timely resolution of such problems. PMID- 9861435 TI - [Experienced anesthetists show better reaction to central gas supply dropout--an anesthesia simulator study]. AB - The loss of pipeline pressure in a central gas supply system is a rare but potentially hazardous complication in anaesthesia and critical care. In an anaesthesia simulator study, reactions of 20 anaesthetists to this simulated critical incident were monitored and evaluated. A comparison between novice (n = 10) and experienced anaesthesia residents/consultants (n = 10) determined a significantly quicker and more on-target reaction by the experienced anaesthetists. Unlike older cycle system anaesthesia machines, update anaesthesia ventilators (CICERO EM, Drager, Lubeck) do not permit manual ventilation of a patient in a "closed-system" once pipeline pressure drops to zero. In this highly hazardous event, the patient has to be ventilated by reservoir bag until a sufficient back-up system delivering high inspiratory oxygen concentrations can be installed, because he is otherwise prone to diffusion hypoxia. Installation of mandatory (anaesthesia-machine integrated) back-up systems for respirators without cycle systems would therefore increase patient safety. A general algorithm for loss of pipeline pressure can be described only after a back-up system has been installed. PMID- 9861436 TI - [Parenteral and enteral feeding]. PMID- 9861438 TI - [Diving injury--emergency procedures outside the routine]. PMID- 9861439 TI - Erythrocyte antioxidant systems protect cultured endothelial cells against oxidant damage. AB - A study was undertaken to assess the ability of the erythrocyte to protect other tissues against oxidative damage. Radiolabelled (51Cr) human umbilical vein endothelial cells (HUVEC) were incubated with erythrocytes and neutrophils activated with phorbol myristate acetate (PMA). Damage to the endothelial cells was indicated by release of radioactivity into the suspending medium. We found that the co-incubation of HUVEC with an increasing range of erythrocyte concentrations resulted in a dose-dependent reduction in the release of radioactivity. When the ability of superoxide to cross the erythrocyte membrane or the glutathione systems was inhibited, the extent of endothelial cell damage increased. Inhibition of the catalase system did not affect results. It was concluded that the erythrocytes afforded some protection against oxidative damage to the endothelial cells by taking up and deactivating the superoxide ions. This protection depends upon intact erythrocyte antioxidant systems. These data support the hypothesis that erythrocytes can provide antioxidant protection to other tissues in vivo. PMID- 9861437 TI - [Dissociative stupor--differential diagnosis of coma following injury]. AB - Two cases are described in which a dissociative stupor originating from conversion neurosis simulated a coma following a sustained trauma. At first both patients showed no response to being addressed or to pain stimuli. They presented an upward eye gaze deviation, cardiorespiratory functions were stable. Following extensive diagnostic procedures revealing no organic cause for the clinical symptoms, the diagnosis of a hysterical consciousness disorder was stated. Symptoms of conversion neuroses include lacking call response, gait disorder, seizure-like conditions and strength diminution in one or more extremities. In these cases suspicious facts are the absence of injuries (for example by falling down or tongue bite during a dissociative attack), eye gaze deviation and the phenomenon that, when the patient's arm is raised above the head and let fall, it never hits the face but glides down beside the body. PMID- 9861440 TI - Changes of glycosylation of serum proteins in psoriatic arthritis, studied by enzyme-linked lectin assay (ELLA), using concanavalin A. AB - Changes of glycosylation of serum proteins of patients with psoriatic arthritis were detected by lectin blotting and a new enzyme-linked lectin assay (ELLA) using concanavalin A (Con A). A good linear correlation was found between the total Con A-reactivity of serum and the serum levels of C-reactive protein and interleukin-6, which is known to regulate the glycosylation pattern of proteins upon inflammation. A good linear correlation was also observed between the immunoreactivity of alpha 1-antitrypsin, measured by ELISA, using a monoclonal antibody sensitive to glycosylation changes, and the erythrocyte sedimentation rate and the serum concentrations of soluble interleukin-2 receptor, an index of lymphocyte activation which correlated with some inflammatory parameters of disease activity. These protein changes, which are described here for the first time, deserve to be studied in further detail in view of their possible clinical applications. PMID- 9861441 TI - Endothelin 1 action on isolated rat stomach and the role of calcium ions in ET 1 induced depolarization of smooth muscle cells BC3H1. AB - Endothelins; (ET1, ET2 and ET3) are a family of peptides which acts on different smooth muscle preparations inducing a slow long lasting contraction. We investigated the effects of ET 1 modulatory action on adrenergic, cholinergic and serotoninergic transmission on an isolated mouse's stomach with gastric nerves. The endothelin 1 stimulation of the mouse stomach tone was abolished by the specific serotonin antagonist methizergid. This study suggests that endothelin 1 plays a role in the regulation of nonvascular smooth muscle tone. The endothelin effect was dependent on free intracellular Ca++ which can be recruited from an extracellular solution as well as from intracellular stores. Complete reduction of Ca++ from the extracellular solution with a simultaneous depletion of calcium stores abolished endothelin 1 depolarization of BC3H1 cells. PMID- 9861442 TI - The effect of heparin and pentosan polysulfate on the thermal stability of yeast alcohol dehydrogenase. AB - Heparin and pentosan polysulfate as organic polyanions inhibit yeast alcohol dehydrogenase (YADH). The aim of this study was to determine the effect of heparin and pentosan polysulfate on the thermostability of alcohol dehydrogenase. Spectral and kinetic analyses showed that these compounds increase the thermal stability of the enzyme and eliminate entirely thermal aggregation. The thermostabilizing effect of unfractionated heparin and pentosan polysulfate was accelerated in the presence of NAD+. The addition of NAD+ (11 microM) to the incubation medium decreased the inhibition of the YADH activity in the presence of pentosan polysulfate (1.32 microM). Moreover, 38% of the residual activity of YADH was found after a 5-min incubation at 70 degrees C. These findings indicate that heparinoids not only modulate the enzyme activity but also can prevent the protein's thermal denaturation. PMID- 9861443 TI - Relationship between rate and extent of catechin absorption and plasma antioxidant status. AB - Flavonoids are described to exert a large array of biological activities, which are mostly ascribed to their radical-scavenging, metal chelating and enzyme modulation ability. Most of these evidences have been obtained by in vitro studies on individual compounds and at doses largely exceeding those dietary. Little is known about a possible relationship between rate and extent of the absorption and modifications of plasma antioxidants. To elucidate this aspect, human volunteers were supplemented with single doses of green tea catechins in free (Greenselect) or phospholipid complex form (Greenselect Phytosome) equivalent to 400 mg epigallocatechingallate (EGCg). EGCg was chosen as biomarker for green tea catechin absorption, and its time course plasma concentration was correlated to the subsequent percent variations of plasma ascorbate, total glutathione, alpha-tocopherol, beta-carotene and Total Radical Antioxidant Parameter (TRAP). Green tea catechins were absorbed more extensively when administered as phospholipid complex rather than as free catechins. Single dose intake of both forms of catechins produced a transient decrease (10-20%) of plasma ascorbate and total glutathione and an increase of plasma TRAP (16-19%). These variations were consistent with the plasmatic levels of EGCg, ascorbate and total glutathione. PMID- 9861444 TI - Sequence-specific DNA binding activity in the RAE28 protein, a mouse homologue of the Drosophila polyhomeotic protein. AB - The rae28 gene, a mouse homologue of the Drosophila polyhomeotic gene, is involved in the maintenance of the transcriptional repression states of Hox genes. In this study we synthesized the glutathione S transferase-RAE28 (GST RAE28) fusion protein and examined sequence-specific DNA binding activity in the RAE28 protein by using the selected and amplified binding site method. After five rounds of enrichment, the eluted DNAs were amplified, cloned and sequenced. The sequences of individual oligonucleotides included the following consensus sequences; 5'-ACCA-3', 5'-ACCCA-3', 5'-CTATCA-3' and 5'-TGCC-3'. The oligonucleotides including these consensus sequences were show to have significant affinity with the GST-RAE28 fusion protein. The RAE28 protein was recently shown to form multimeric protein complexes with other members of mouse Pc-G proteins in the nucleus. These findings strongly suggest that the RAE28 protein constitutes a sequence-specific DNA binding domain in multimeric Pc-G protein complexes. PMID- 9861446 TI - HMG CoA reductase inhibitor accelerates aging effect on diaphragm mitochondrial respiratory function in rats. AB - We examined effects of pravastatin on age-related changes in mitochondrial function in rats. Decline in the activity of complex I of the mitochondrial electron transport chain was observed in diaphragm and psoai major in rats aged 35 and 55 weeks, and that of complex IV in rats aged 55 weeks. Pravastatin accelerated significantly age-related decline in the activity of complex I of diaphragm mitochondria, though pravastatin did not show significant effect on normally observed age-associated decline in the activities of complex IV of psoai major and diaphragm mitochondria. Aging effect on mitochondrial respiratory function was not observed on heart muscle and liver in rats up to 55 weeks old, and pravastatin did not effect significantly heart and liver mitochondrial respiratory function. From these results, careful clinical examination on respiratory muscle function should be necessary in patients treated with pravastatin particularly in elderly patients. PMID- 9861445 TI - Amino acid residues in third intracellular loop of melanocortin 1 receptor are involved in G-protein coupling. AB - To delineate domains essential for G-protein coupling in melanocortin 1 receptor (MC1R), we mutated polar and basic residues to alanine at eleven positions in the putative third intracellular loop and determined consequent changes in the ligand binding and generation of second messenger cAMP. Results demonstrate that ligand binding affinity was not affected by any of the mutations. However, every mutant displayed reduced functional response as compared to the wild type receptor. Replacement of residues (K226, R227, Q228, R229, H232, Q233 and K238) present in second half of third intracellular loop resulted in an almost complete loss of functional response. The results have demonstrated that the amino acid residues present in C-terminal portion of third intracellular loop of MC1R are involved in coupling to G-protein and that a region of four amino acids, K226-R227-Q228-R229 is essential for coupling of MC1R to G-protein. PMID- 9861447 TI - Protein binding in vivo to OP2 promoter of the Pseudomonas putida TOL plasmid. AB - The transcription of OP2 encoding enzymes for m-toluate catabolism on the Pseudomonas putida TOL plasmid is activated by basal-level XylS protein in the presence of m-toluate or by overproduced XylS protein in the absence of m toluate. In this study, in vivo dimethyl sulfate (DMS) footprinting was performed to understand the mechanism of transcriptional regulation of OP2 promoter by XylS. In the presence of overproduced XylS without m-toluate, several protected nucleotides were observed, indicating the binding of RNA polymerase to DNA. However, the protection was canceled upon addition of m-toluate. These results suggest that RNA polymerase is retained by XylS on the OP2 promoter in the absence of inducer, and is released by m-toluate binding to XylS, concomitant with transcription. PMID- 9861448 TI - Interactions of photosensitized tetracycline with serum albumin. AB - Interactions of tetracycline with bovine serum albumin (BSA) were studied by fluorescence quenching and circular dichroism (CD) analysis. The binding isotherm exhibited at least 13 tetracycline binding sites on the albumin molecule. Amongst these, four were found to be high affinity sites and the remainder were loose sites. The Scatchard analysis demonstrated the binding constant and capacity of BSA to be 4.6 x 10(6) liters/mole and 3.6, respectively. The CD data revealed a significant decrease in the mean residue ellipticity (MRE), indicating alterations in the protein helicity. A reduction of 20% in the alpha-helical content of the albumin was noted at higher levels of tetracycline in the presence of Cu (II) ions. Thus the strong in vitro interactions of tetracycline with albumin resulted in conformational changes in its globular structure and insinuate potential health risk due to possible macromolecular damage, under physiological conditions, from the formation of tetracycline/Cu(II) complexes. PMID- 9861449 TI - The plasma membrane Fe(3+)-reductase activity of Caco-2 cells is modulated during differentiation. AB - The aim of the present study was to investigate whether the brush border membrane ferric reductase activity of Caco-2 cells is modulated during cell differentiation. The ferric reductase activity was determined in whole cells and isolated microvillous membranes at different stages of cell differentiation by measuring the amount of Fe3+ reduced during the incubation time. Our results indicated that the ferric reductase activity decreased in fastly growing cells and reactivated in postconfluent cells in contrast to the alkaline phosphatase and sucrase activities which were progressively expressed during differentiation as conventional indicators of cell maturity. The lowest ferric reductase activity was found in cells at the log phase of proliferation, while freshly seeded or highly differentiated cells had significantly higher enzyme activities. Cells grown under serum-free conditions had similar ferric iron reduction rates as cells propagated under standard conditions. Reagents or hormones affecting cell metabolism through different pathways had no significant effect on this transplasma membrane redox system. PMID- 9861450 TI - Effect of chronic alcohol ingestion on buccal mucosal expression of bFGF and Cdk2 during ulcer healing. AB - In this study, we investigated the effect of chronic alcohol ingestion on the interplay between the receptor-bound basic fibroblast growth factor (bFGF-R) and the expression of cyclin-dependent kinase (Cdk2) in buccal mucosa during ulcer healing. Chronic ulceration was induced by a topical application of acetic acid to the buccal mucosa of rats maintained for 5 weeks on alcohol-containing or control liquid diet. In both groups, the ulcer healing was accompanied by an increase in buccal mucosal expression of bFGF and Cdk2. In the control group, the ulcer healed within 10 days and maximum induction in bFGF (2.6-fold) and Cdk2 (2.4-fold) occurred by the 2nd day of healing. In contrast, the alcohol diet group showed a marked delay in ulcer healing (14 days), associated with the shift in maximum of bFGF and Cdk2 expression to the 4-6th day, and the values were reduced by 35 to 38%. The findings show that chronic alcohol ingestion exerts detrimental effect on the signaling events initiated by bFGF-receptor activation and propagated by Cdk2 that propels the cell cycle progression essential for rapid mucosal repair. PMID- 9861451 TI - Different sources of acidity in glucose-elicited extracellular acidification in the yeast Saccharomyces cerevisiae. AB - Three wild-type strains of Saccharomyces cerevisiae, viz. K, Y55 and sigma 1278b, two mutants lacking one or both of the putative K+ transporters, trk1 delta and trk1 delta trk2 delta, and a mutant in the plasma membrane H(+)-ATPase, viz. pma1 105, were compared in their extracellular acidification following addition of glucose and subsequent addition of KCl; in ATPase activity in purified plasma membranes; and in respiration on glucose. The glucose-induced acidification was the greater the higher the respiratory quotient, i.e. the higher the anaerobic metabolism. A markedly lower acidification was found in the ATPase-deficient pma1 105 strain but also in the TRK-deficient double mutant. The acidification pattern after addition of KCl corresponds to expectations in the TRK mutants; however, a similarly decreased acid production was found in the ATPase-deficient mutant pma1 105. The highest rate of ATP hydrolysis in vitro was found with the trk1 delta trk2 delta mutant where glucose-, as well as KCl-induced acidification were lowest. Likewise, the pma1-105 mutant with extremely low acidification showed only a minutely lower ATP hydrolysis than did its parent Y55 strain. Apparently, several different sources of acidity are involved in the glucose-induced acidification (including extrusion of organic acids); in fact, contrary to the general belief, the H(+)-ATPase may play a minor role in this process in some strains. PMID- 9861452 TI - Inhibition of wheat leaves nitrate reductase activity by cibacron blue. AB - Cibacron Blue F3GA (CB) inhibited the activities of wheat leaves NADH:nitrate reductase and NADH:cytochrome-c reductase in a time-independent and concentration dependent manner. The methyl viologen:nitrate reductase activity of the enzyme was unaffected by various CB concentrations used in the experiment. Inhibition of NADH:nitrate reductase was of mixed type (partial competitive and pure noncompetitive) with respect to NADH and noncompetitive with respect to nitrate. The estimated inhibition constant (Ki) values were 1 microM for NADH and 8.4 microM for nitrate. The secondary plots of inhibition with respect to NADH, indicated a dissociation constant (KI) of 8.8 microM for the enzyme-NADH-CB complex. This KI being greater than the Ki suggested that the noncompetitive inhibition is predominant over the competitive inhibition at the NADH binding site. PMID- 9861453 TI - Evaluation of 9-cis retinoic acid for a new remedy of human retinoblastoma. AB - We investigated the effect of two isomers of retinoic acid (RA), all-trans RA and 9-cis RA, on the proliferation of Y79 human retinoblastoma cells. The two isomers inhibited the cell proliferation in a concentration-dependent manner. The IC50 for this inhibition by all-trans RA and 9-cis RA was 1.50 and 0.15 microM, respectively. The inhibitory effect of 9-cis RA on Y79 cell growth was observed within 24 hr, thereafter the cell number was gradually decreased. In contrast, no inhibition by all-trans RA of Y79 cell growth was observed within 24 hr, thereafter the cell number was slightly increased. In these cases, the cell viability at 4 days after the addition of 9-cis RA and all-trans RA was more than 90% and 95%, respectively. These results indicate that the two RA inhibit the proliferation of Y79 human retinoblastoma cells without inducing the cell death and that the effect of 9-cis RA on the inhibition of Y79 cell growth is much greater than that of all-trans RA. PMID- 9861454 TI - Sphingosine induces phospholipase D and mitogen activated protein kinase in vascular smooth muscle cells. AB - The enzymes phospholipase D and diacylglycerol kinase generate phosphatidic acid which is considered to be a mitogen. Here we report that sphingosine produced a significant amount of phosphatidic acid in vascular smooth muscle cells from the rat aorta. The diacylglycerol kinase inhibitor R59 949 partially depressed sphingosine induced phosphatidic acid formation, suggesting that activation of phospholipase C and diacylglycerol kinase can not account for the bulk of phosphatidic acid produced and that additional pathways such as phospholipase D may contribute to this. Further, we have shown that phosphatidylethanol was produced by sphingosine when vascular smooth muscle cells were stimulated in the presence of ethanol. Finally, as previously shown for other cell types, sphingosine stimulated mitogen-activated protein kinase in vascular smooth muscle cells. PMID- 9861455 TI - Effects of prostaglandins and nitric oxide on rat macrophage lipid metabolism in culture: implications for arterial wall-leukocyte interplay in atherosclerosis. AB - Macrophages/foam cells have a pivotal role in atherogenesis although little is known about the way lipid imbalance, a hallmark of atherosclerosis, leads to lipid accumulation in these cells. Modified low-density lipoproteins are associated with macrophage lipid dysfunction in atherosclerosis, but a possible role for altered lipogenesis leading to lipid accumulation remains to be elucidated. Since endothelium-derived nitric oxide (NO) and prostaglandins (PGs) are physiological autacoids whose production may be impaired in atherosclerosis, the effects of these mediators on de novo lipid synthesis in 24-h cultured rat peritoneal macrophages is investigated. In resident (unstimulated) cells, 1 microM PGE2 and the stable analog of PGI2 carbaprostacyclin (cPGI2, 1 microM) deviated the overall [1-14C]acetate from incorporation into cholesterol, free fatty acids and triacylglycerols favoring the formation of phospholipids. In inflammatory (thioglycollate-elicited) macrophages, these eicosanoids likewise reduced 14C-incorporations into all the lipid fractions tested. Also, cPGI2 and PGE2 reduced [4-14C]cholesterol uptake from inflammatory cells but did not interfere in 14C-cholesterol export. The PGE2-derivative PGA2 (10-20 microM) reduced 14C-incorporations into all the lipids in resident cells while it enhanced phospholipid synthesis by up to 129% at the expense of reduced incorporations into the other test lipids. The NO donor S-nitroso-N acetylpenicillamine (SNAP, 1-10 microM), when added to macrophages in the presence of superoxide dismutase (SOD, to avoid the reaction of superoxide with NO), significantly reduced lipogenesis especially in inflammatory cells. These findings suggest that endothelium-derived NO and PGs may be associated with macrophage lipid accumulation by modulating lipogenesis and cholesterol uptake within these cells. PMID- 9861456 TI - Suppression of constitutive and inducible cytochrome P450 gene expression by alpha-hederin in mice. AB - The effects of alpha-Hederin, a triterpenoid saponin which exists in some oriental herbs, on the expression of liver cytochrome P450s were examined in mice. The administration of alpha-Hederin to mice significantly decreased the hepatic content of P450 and the activities of microsomal ethoxyresorufin O deethylase, methoxyresorufin O-demethylase, and aniline hydroxylase, representative activities of cytochrome-P4501A1, -P4501A2, and -P4502E1, respectively, in a dose- and time-dependent manner. However, pentoxyresorufin O dealkylase, a representative activity of cytochrome P4502B1/2, was decreased to a lesser extent. alpha-Hederin also decreased inducible monooxygenase activities in the same manner. Suppressions of P450 isozyme expression occurred in alpha Hederin treated hepatic microsomes, as determined by immunoblot analysis in a manner consistent with that of the enzyme activity levels. Levels of mRNA of P4501A1/2 and P4502B1/2 were also decreased by alpha-Hederin as shown by Northern blot analysis. In contrast, the level of P4502E1 mRNA in the liver of alpha Hederin treated mice was unchanged. These results suggest that alpha-Hederin may act as a more specific suppressor for P4501A and P4502E1 than P4502B and that the suppression involves decreases in mRNA levels except in the case of P4502E1. PMID- 9861457 TI - A six-domain structural model for Escherichia coli translation initiation factor IF2. Characterisation of twelve surface epitopes. AB - The Escherichia coli translation initiation factor IF2 is a 97 kDa protein which interacts with the initiator fMet-tRNAfMet, GTP and the ribosomal subunits during initiation of protein biosynthesis. For structural and functional investigations of the factor, we have raised and characterised monoclonal antibodies against E. coli IF2. Twelve epitopes have been localised at the surface of the protein molecule by three different methods: Interactions of the monoclonal antibodies with nested deletion mutants of IF2, comparison of the relative location of the epitopes in a competition immunoassay and cross-reactivity analyses of the monoclonal antibodies towards IF2 from Salmonella typhimurium, Klebsiella oxytoca, Enterobacter cloacae, Proteus vulgaris, and Bacillus stearothermophilus. These data are combined with predicted secondary structure and discussed in relation to a six-domain structural model for IF2. The model describes IF2 as a slightly elongated molecule with a structurally compact C-terminal domain, a well conserved central GTP-binding domain, and a highly charged, solvent exposed N terminal with protruding alpha-helical structures. PMID- 9861458 TI - Identification and partial characterization of three calcium- and zinc independent gelatinases constitutively present in human circulation. AB - Three constitutive gelatinases in human plasma were identified and characterized relative to known matrix metalloproteinase (MMP) gelatinases: MMP-2 (fibroblast 72-kDa) and MMP-9 (neutrophil 92-, 130-, and 225-kDa). Substrate gel electrophoresis (gelatin zymography) revealed an apparent Mw of 78-, 82-, and 89 kDa for these gelatinases. Densitometry revealed that MMP-9 and MMP-2 were highly calcium sensitive requiring 50-150 microM and 500 microM calcium for half-maximal activity, respectively. Of the new gelatinases, only the 89-kDa form demonstrated slight calcium activation. The three gelatinases were unaffected by known MMP inhibitors: EDTA (5 mM), 1,10-phenanthroline (2 mM), and pepstatin (18 microM). Serine and thiol protease inhibitors (leupeptin, aprotinin, PMSF, TLCK, TPCK, antichymostatin, antipain) were also ineffective. Solution-phase IEF revealed that the 78- and 82-kDa forms focused at neutral pI 6.72-7.95 whereas the 89-kDa focused at an acidic pI 4.89-5.18 (similar to neutrophil and fibroblast forms). The data indicate that these gelatinases are not MMPs or partially activated MMPs. Their role in normal and pathological conditions is not known. PMID- 9861459 TI - Differences in ethidium bromide and 4'-6-diamidino-2-phenylindole staining profiles with regard to DNA fragmentation during apoptosis. AB - To simply and directly evaluate DNA fragmentation during apoptosis induced in mouse cultured hepatocytes by an anti-Fas antibody, we examined the fluorescence intensity in cell nuclei stained with ethidium bromide and 4'-6-diamidino-2 phenylindole by optiphoto fluorescence microscopy. The intensity of the former staining for the nuclear DNA of apoptotic cells was clearly decreased compared to that of non-apoptotic cells, whereas no difference in the fluorescence intensity for the latter stain between the apoptotic and non-apoptotic groups was observed. Thus, the use of optiphoto fluorescence microscopy, in conjunction with both stains, constitutes a useful tool for the evaluation of apoptotic DNA fragmentation. PMID- 9861461 TI - Brain neurocircuitry of anxiety and fear: implications for clinical research and practice. PMID- 9861460 TI - Induction of caspase-3 and nitric oxide synthase-2 during gastric mucosal inflammatory reaction to Helicobacter pylori lipopolysaccharide. AB - Helicobacter pylori lipopolysaccharide is recognized as a primary virulence factor evoking acute mucosal inflammatory reaction associated with H. pylori infection. We investigated the activity of a key apoptotic protease, caspase-3, and the expression of inducible nitric oxide synthase (NOS-2) during H. pylori lipopolysaccharide-induced acute gastritis. The assays conducted 4 days following intragastric dose of the lipopolysaccharide revealed a pattern of acute mucosal responses characterized by an 11.2-fold increase in epithelial cells apoptosis, inflammatory infiltration of the lamina propria, hyperemia, and epithelial hemorrhage. This was accompanied by a 5.4-fold increase in caspase-3 activity, while the mucosal expression of NOS-2 showed a 6.5-fold induction. The results implicate H. pylori lipopolysaccharide in the induction of NOS-2 expression, and point to its effect on activation of the signaling cascade involving caspase-3 in the process gastric epithelial cells apoptosis. PMID- 9861462 TI - Translational implications of the amygdala-stria terminalis model for the clinical anxiety disorders. PMID- 9861463 TI - Binge-eating and bulimia: potential insights into etiology and pathophysiology through genetic epidemiologic studies. PMID- 9861464 TI - Heritability of binge-eating and broadly defined bulimia nervosa. AB - BACKGROUND: Using diagnostic information obtained at two different times, we incorporated error of measurement into structural equation twin models to evaluate the contribution of additive genetic, common environmental, and individual-specific environmental factors to the liability to binge-eating and broadly defined bulimia nervosa (BN). We also evaluated the validity of the equal environment assumption (EEA) with reference to these two phenotypes. METHODS: We interviewed 1897 female twins (including both members of 854 twin pairs) from a population-based register about their lifetime history of binge-eating and of broadly defined BN twice, approximately 5 years apart. RESULTS: The reliabilities of a lifetime history of binge-eating (kappa = .34) and of broadly defined BN (kappa = .28) were low. Based on single interviews, the heritability of binge eating was estimated to be 50% and broad BN 60%, with the remaining variance attributable to individual-specific environment. Common environmental influences had no effect on liability to either trait. By combining information from two interview waves and thereby incorporating error of measurement into a structural equation model, the estimated heritability of the latent vulnerability to binge eating (82%) and broadly defined BN (83%) increased substantially. Although there were no violations of the EEA detected for binge-eating, cosocialization influenced twin concordance for broadly defined BN. CONCLUSIONS: Lifetime histories of binge-eating and broadly defined BN appear to be highly heritable conditions of low reliability. PMID- 9861465 TI - The emotional counting Stroop paradigm: a functional magnetic resonance imaging probe of the anterior cingulate affective division. AB - BACKGROUND: The emotional counting Stroop (ecStroop) functional magnetic resonance imaging (fMRI) activation paradigm was designed to recruit the anterior cingulate affective division (ACad). METHODS: Nine normal, healthy male and female subjects (mean age 24.2 years) reported via button press the number of neutral and negative words that appeared on a screen while reaction time and fMRI data were acquired. RESULTS: We observed a) greater ACad activation for negative versus neutral words during initial presentation blocks; b) lower overall ACad signal intensity during task performance (i.e., both negative and neutral words) compared to the baseline fixation condition; and c) no reaction time increase to negative versus neutral words. CONCLUSIONS: In a companion study of a cognitive version of the counting Stroop (Bush et al 1998), these same 9 subjects a) activated the more dorsal anterior cingulate cognitive division; b) also showed the overall decrease in ACad signal intensity; and c) demonstrated a reliable reaction time effect. Taken together, these data offer a within-group spatial dissociation of AC function based upon information content (i.e., cognitive vs. emotional) and/or presence of behavioral interference. We propose that the ecStroop will be a useful fMRI probe of ACad function in anxiety disorders. PMID- 9861466 TI - Fear and the brain: where have we been, and where are we going? AB - In recent years, there has been an explosion of interest in the neural basis of emotion. Much of this enthusiasm has been triggered by studies of the amygdala and its contribution to fear. This work has shown that the amygdala detects and organizes responses to natural dangers (like predators) and learns about novel threats and the stimuli that predict their occurrence. The latter process has been studied extensively using a procedure called classical fear conditioning. This article surveys the progress that has been made in understanding the neural basis of fear and its implications for anxiety disorders, as well as the gaps in our knowledge. PMID- 9861467 TI - Are different parts of the extended amygdala involved in fear versus anxiety? AB - Although there is a close correspondence between fear and anxiety, and the study of fear in animals has been extremely valuable for understanding brain systems that are important for anxiety, it is equally clear that a richer animal model of human anxiety disorders would include measures of both stimulus-specific fear and something less stimulus specific, more akin to anxiety. Studies in patients with posttraumatic stress syndrome indicate these individuals seem to show normal fear reactions but abnormal anxiety measured with the acoustic startle reflex. Studies in rats, also using the startle reflex, indicate that highly processed explicit cue information (lights, tones, touch) activates the central nucleus of the amygdala, which in turn activates hypothalamic and brain stem target areas involved in specific signs of fear. Somewhat less explicit information, such as that produced by exposure to a threating environment for several minutes or by intraventricular administration of the peptide corticotropin-releasing hormone may activate a brain area closely related to the amygdala, called the bed nucleus of the stria terminalis, which in turn activates hypothalamic and brain stem target areas involved in specific signs of fear or anxiety. Because the nature of this information may be less specific than that produced by an explicit cue, and of much longer duration, activation of the bed nucleus of the stria terminalis may be more akin to anxiety than to fear. PMID- 9861468 TI - Emotion, motivation, and anxiety: brain mechanisms and psychophysiology. AB - The organization of response systems in emotion is founded on two basic motive systems, appetitive and defensive. The subcortical and deep cortical structures that determine primary motivated behavior are similar across mammalian species. Animal research has illuminated these neural systems and defined their reflex outputs. Although motivated behavior is more complex and varied in humans, the simpler underlying response patterns persist in affective expression. These basic phenomena are elucidated here in the context of affective perception. Thus, the research examines human beings watching uniquely human stimuli--primarily picture media (but also words and sounds) that prompt emotional arousal--showing how the underlying motivational structure is apparent in the organization of visceral and behavioral responses, in the priming of simple reflexes, and in the reentrant processing of these symbolic representations in the sensory cortex. Implications of the work for understanding pathological emotional states are discussed, emphasizing research on psychopathy and the anxiety disorders. PMID- 9861469 TI - Brain circuits in panic disorder. AB - This paper reviews the pathophysiology of panic disorder (PD), within the context of newly described "fear circuitries," which have been well characterized in preclinical models. Substantial advances in the neurosciences have made it possible for clinical neuroscientists to refine our understanding of the pathophysiology of PD and the mechanisms of currently effective treatment. These advances have in turn helped generate testable hypotheses for future neurobiological and psychopharmacologic research. Perturbation of mutual modulation ("cross talk") between key brain transmitter systems (serotonin, norepinephrine, gamma-aminobutyric acid, corticotropin-releasing factor, and others) may underlie the pathogenesis of panic-anxiety. Restoration of normal homeostasis may be an important therapeutic component of antipanic therapy and may provide information about underlying neurocircuits. Neuroimaging, an important new tool, has already begun to bridge the gap between the preclinical and clinical neurosciences through confirmation of hypothesized dysfunction of the complex human prefrontal cortex and its subcortical components. In higher species, such as humans, dysfunction of cortical inhibition or excessive cortical activation of caudal limbic structures is postulated to lead to activation of the phylogenetically conserved amygdalofugal pathways. Consistent with probable subtypes of PD, overlapping theoretical models of panic neurocircuitries are proposed, including ventilatory dysregulation, which is coupled with neurovascular instability in a critical area of the panic neurocircuitry--the amygdalohippocampus. Neuroimaging appears a critical tool in guiding further elaboration of the interaction of cortical and subcortical components of the panic neurocircuitry, whereas challenge studies appear crucial in gathering further information regarding brain stem dysfunction. PMID- 9861470 TI - Neurobiological perspectives on social phobia: from affiliation to zoology. AB - Social phobia (or "social anxiety disorder") is a prevalent condition that has been the subject of increased scrutiny in recent years. The purpose of this paper is to review the neurobiology of social phobia. It is apparent from the extant literature that this disorder is poorly understood from a neurobiological perspective. There are nonetheless a number of clinical and preclinical observations which, at times, converge to illuminate areas worthy of further study. Included in this category are suggestive findings of central serotonergic dysregulation in social phobia, response to serotonin reuptake inhibitors in social phobia, and the role of serotonergic function in septohippocampal models of anxiety. Abnormalities in central dopaminergic function are also posited, supported to some extent by recent neuroimaging findings. There are in addition a number of animal and human behavioral models in existence that may be relevant to the study of social phobia. Included in this category are models of social dominance in wild baboons, social affiliation in the prairie vole, and behavioral inhibition to the unfamiliar in childhood. Newer technologies that are likely to play a major role in the delineation of the neural circuitry (e.g., functional magnetic resonance imaging) and heritability (e.g., molecular genetics) of social phobia are discussed. Finally, an interactive role for biology and experience in the expression of social phobia is considered. PMID- 9861471 TI - Generalized anxiety disorder: neurobiological and pharmacotherapeutic perspectives. AB - The concept of generalized anxiety has evolved over many years, from initial descriptions of "anxiety neurosis" to recognition of generalized anxiety disorder (GAD) as a clinical entity included in the 3rd edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-III) in 1980. Since 1980, the definition of GAD has undergone further change, with modifications in the salience of autonomic and panic like symptoms, duration, and allowance of comorbidity. The importance of these historical considerations lies in the fact that most of our current knowledge about GAD rests on outdated definitions, including most of the literature pertaining to treatment. Indeed, we cannot be sure that the DSM-III definition of GAD bears resemblance to the current concept, and these differences may have profound implications for findings from research. The following two-part report comprises i) a review of the basic neurobiology of GAD, with reference to serotonergic, noradrenergic, neuroendocrine, autonomic imaging, and other systems; and ii) an overview of the current status of pharmacotherapy for GAD. PMID- 9861472 TI - Current approaches to etiology and pathophysiology of specific phobia. AB - Specific phobia is a common, heterogeneous disorder whose central feature is persistent, unreasonable fear of a circumscribed object or situation. This article reviews current etiological theories and empirical data that seem likely to be important in investigating the pathophysiology of this disorder. These include conditioning, modified conditioning, and nonassociative models of phobia development, physiological response to the phobic stimulus, neuroimaging, primate, and biological challenge studies. Pathophysiological hypotheses suggested by recent research on the neurocircuitry of conditioned fear are also discussed. Though specific phobias have been of less public health and clinical interest than other anxiety disorders, their circumscribed nature and possible relationship to conditioned fear may make them a productive subject for research into basic pathophysiology. PMID- 9861473 TI - Predicting the development of posttraumatic stress disorder from the acute response to a traumatic event. AB - Posttraumatic stress disorder (PTSD) is a psychiatric condition that is directly precipitated by an event that threatens a person's life or physical integrity and that invokes a response of fear, helplessness, or horror. In recent years it has become clear that only a proportion of those exposed to fear-producing events develop or sustain PTSD. Thus, it seems that an important challenge is to elucidate aberrations in the normal fear response that might precipitate trauma related psychiatric disorder. This paper summarizes the findings from recent studies that examined the acute and longer term biological response to traumatic stress in people appearing to the emergency room immediately following trauma exposure. In the aggregate, these studies have demonstrated increased heart rate and lower cortisol levels at the time of the traumatic event in those who have PTSD at a follow-up time compared to those who do not. In contrast, certain features associated with PTSD, such as intrusive symptoms and exaggerated startle responses, are only manifest weeks after the trauma. The findings suggest that the development of PTSD may be facilitated by an atypical biological response in the immediate aftermath of a traumatic event, which in turn leads to a maladaptive psychological state. PMID- 9861474 TI - Shortened REM latency as a psychobiological marker for psychotic depression? An age-, gender-, and polarity-controlled study. AB - BACKGROUND: Previous reports suggest that the clinical dichotomy separating psychotic and nonpsychotic depression corresponds to different neurobiological profiles. The aim of the present study is to further investigate the psychobiological correlates of these two particular depressive subtypes. METHODS: Thyroid-stimulating hormone response to thyrotropin-releasing hormone postdexamethasone cortisol levels, and electroencephalgraphic sleep characteristics of 44 psychotic major depressive patients were compared to those of 44 nonpsychotic depressives matched for age, gender, and polarity. RESULTS: Some biological disturbances usually associated with depression (increased wakefulness, diminished rapid eye movement latency, hypercortisolism, blunted thyroid-stimulating hormone response to thyrotropin-releasing hormone stimulation) seemed to be significantly more pronounced in the psychotic depressed group as a reflection of greater illness severity; however, shortened REM latency was not influenced by severity and seemed to be more specifically related to the co-occurrence of psychotic and depressive symptoms. CONCLUSIONS: Our data provide further support for the validity of the clinical dichotomy separating psychotic and nonpsychotic major depression independently of severity. PMID- 9861475 TI - Cerebrospinal fluid monoamines in Prader-Willi syndrome. AB - BACKGROUND: The behavioral phenotype of Prader-Willi syndrome (PWS) suggests hypothalamic dysfunction and altered neurotransmitter regulation. The purpose of this study was to examine whether there was any difference in the concentrations of monoamine metabolites in the cerebrospinal fluid (CSF) in PWS and non-PWS comparison cases. METHODS: The concentration of monoamine metabolites in CSF was determined in 13 children and adolescents with PWS diagnosed on clinical and genetic criteria. The concentrations were compared with those from 56 comparison cases in healthy and other contrast groups. RESULTS: The concentrations of dopamine and particularly serotonin metabolites were increased in the PWS group. The differences were most prominent for 5-hydroxyindoleacetic acid. The increased concentrations were found in all PWS cases independently of age, body mass index, and level of mental retardation. CONCLUSIONS: The findings implicate dysfunction of the serotonergic system and possibly also of the dopamine system in PWS individuals, and might help inform future psychopharmacologic studies. PMID- 9861476 TI - Prefrontal and striatal dopamine metabolism during enhanced rebound hyperphagia induced by space restriction--a rat model of binge eating. AB - BACKGROUND: Several lines of evidence indicate that abnormalities in brain dopamine and serotonin metabolism may play an important role in bulimia nervosa. However, the regional neurochemical mechanism of the binge eating is poorly understood. Our purpose was to elucidate brain neurochemical mechanisms of binge eating using a rat model. METHODS: The dopamine release and metabolism in the prefrontal cortex (PFC) and in the ventrolateral striatum (VLS) of rats were studied using microdialysis during enhanced rebound hyperphagia induced by space restriction (an animal model of binge eating). RESULTS: The rats showed rebound hyperphagic state when they were released from scheduled feeding (2 hours/day feeding for 7 days). The hyperphagia was further enhanced when they were put in a space-restricted cage where their mobility was restricted. Dopamine release and metabolism were increased both in the PFC and in the VLS during the enhanced rebound hyperphagia. CONCLUSIONS: These results tentatively suggest that increased dopamine release and metabolism in the PFC and in the VLS may be related to space restriction and to activation of motor function involved in feeding behavior, respectively. The enhanced rebound hyperphagia induced by space restriction may be useful as an animal model of binge eating. PMID- 9861477 TI - The temporal dynamics of tics in Gilles de la Tourette syndrome. AB - BACKGROUND: Statistical characterization of tic behavior in Gilles de la Tourette syndrome (GTS) may provide insight into the dynamic functioning of the human central nervous system, as well as improve the quantitative assessment of tic symptom severity. METHODS: Twenty-two medication-free GTS subjects underwent videotaping of their tics. The intervals between temporally adjacent tics were measured, and the statistical properties of these intervals were assessed through graphical representation of frequency distributions, autoregressive integrated moving average (ARIMA) modeling, spectral analysis, and construction of first return maps. RESULTS: The frequency distribution of tic interval durations followed an inverse power law of temporal scaling. Spectral analyses similarly demonstrated that the spectral power density of tic interval duration scales inversely with frequency. ARIMA modeling suggested that the time series for tics are nonstationary as well as moving average processes. The first return maps demonstrated "burstlike" behavior and short-term periodicity in tics, and proved that successive tic intervals are not statistically independent. Graphic display of the time series confirmed shortterm periodicity, and in addition suggested the presence of period doubling. CONCLUSIONS: These findings are suggestive though not conclusive evidence for the presence of a fractal, deterministic, and possibly chaotic process in the tic time series. These analytic methods provide insight into the temporal features of tics that commonly are described clinically (such as short-term bouts or bursting, and longer term waxing and waning), and they reveal certain important temporal features of tics that have not been clinically described. The methods may also prove useful in the improved characterization of tic symptom severity. PMID- 9861479 TI - Diet and rat strain as factors in nervous system function and influence of confounders. AB - The necessity for understanding normal human cognitive processes and behavior, and the mechanisms which result in dysfunction in these processes are dependent on utilization of a suitable animal model. In order to develop pharmaceutical agents to alleviate mental disturbances and enable the individual to cope within the norms of society, it is incumbent upon investigators to choose a species in which pharmacokinetic principles are established and resemble those of humans. The choice of rats in cognition research studies has specific advantages in that these animals possess similar pharmacodynamic parameters to humans. Further advantages include availability, low cost, ease of breeding, maintenance and an extensive literature database which enable comparisons to present findings. However, there are substantial differences in the performance of various rat strains in tasks of learning, memory, attention, and responses to stress or drugs. In addition to rat strain, quantity of food also exerts profound consequences on animal behavior. The aim of this review is to demonstrate that there are differences in the central nervous system responsiveness of rat strains to chemical and these could be related to factors such as source of supplier, type and quantity of feed, or season of the year. It is also evident that the genotype differs amongst strains and this may be responsible for the observed differences in CNS sensitivity to chemicals. Strain differences must be identified and taken into consideration in interpretation of assessment of neurobehavioral functions. It is also incumbent upon the investigators to utilize healthy (diet-controlled) animal models. PMID- 9861478 TI - Cerebrospinal fluid levels of oxytocin in Prader-Willi syndrome: a preliminary report. AB - BACKGROUND: Prader-Willi syndrome (PWS) is a genetic disorder characterized by mental retardation, appetite dysregulation, and a high risk for obsessive compulsive disorder (OCD). Microscopic abnormalities of the hypothalamus have been described in PWS, and oxytocin has been implicated in both appetite regulation and OCD. METHODS: Oxytocin and arginine vasopressin (AVP) were measured in the cerebrospinal fluid of 5 subjects with PWS (2 male, 3 female) and in 6 normal control subjects (all female). RESULTS: CSF oxytocin was elevated in PWS (9.2 +/- 3.9 pmol/L) as compared to normal control subjects (5.1 +/- 0.9 pmol/L, p = 0.045), a finding that was more significant when excluding male subjects from analysis (p = 0.02). AVP was not significantly different between the groups as a whole. CONCLUSIONS: These data provide further evidence for hypothalamic and oxytocinergic dysfunction in PWS. The associations between oxytocin, appetite regulation, and obsessive compulsive symptomatology in PWS warrant further investigation. PMID- 9861480 TI - Lindane degradation and effects on soil microbial activity. AB - The degradation of U-14C-lindane in two Egyptian soils was determined in a three month laboratory incubation. Lindane mineralization was slow and limited in both soils. Evolution of 14CO2 increased with time but only reached 3.5 to 5.5% of the initial 14C-concentration within 90 days. At that time both soils contained about 88% of the applied radiocarbon; 33% to 37% of the initial dose was unextractable and assumed bound to the soils. The methanol-extractable 14C primarily contained lindane with traces of minor metabolites. Radiorespirometry was used to evaluate the effect of lindane on soil microbial activity. Low concentrations of the insecticide initially supressed 14CO2 evolution from U-14C-glucose and microbial activity was significantly inhibited by 10 mg lindane/kg soil. PMID- 9861481 TI - p53 gene mutations in asbestos associated cancers. AB - The accumulation of mutant p53 protein in cancer cells was observed by immunohistochemistry analysis. DNA was extracted from paraffin-embedded tissue. Exons 5, 7 and 8 were amplified and studied by PCR-SSCP and sequencing analysis. Ten cases of asbestos associated cancer tissue were studied, of which five cases had adenocarcinoma, and the other five had mesothelioma, squamous carcinoma, small cell lung cancer, adenosquamous carcinoma and malignant lymphoma respectively. Employing monoclonal antibody PAb1801, five cases were found to be mutant p53 protein positive. Seven cases were found to have mutations by PCR SSCP. A total of 7 cases (8 mutations) were found to be positive and 4 cases were found to be positive by both of these analyses. Of the 8 mutations found by SSCP analysis, 4(50%, 4/8) were clustered in exon 8. A high mutation frequency was noticed in adenocarcinoma (80%, 4/5). Sequencing analysis on two specimens revealed two hotspot mutations. In codon 234, TAC for tyrosin was mutated to AAC for asparagine by a T to A transversion of the first letter. In codon 273, CGT for arginine was mutated to AGT for serine by a C to A transversion of the first letter. In conclusion, the mutation of p53 gene is common in asbestos associated cancers. However, the mutational spectrum of asbestos associated cancers might be different from that of non-asbestos associated cancers. PMID- 9861482 TI - Effect of protein malnutrition on sex organs of metanil yellow exposed male rats. AB - Oral administration of metanil yellow (MY) at 3.0% (w/w) dose level to adult male albino rats maintained on low protein (LP) diet for 30 days resulted in a greater decrease in absolute and relative weights of testes than in those rats maintained on a normal protein (NP) diet. A marked decrease in the activities of lactate dehydrogenase and hyaluronidase and content of lactic acid in LP + MY fed animals suggested that low protein diet enhanced the vulnerability of germ cells towards metanil yellow. The lack of significant changes in the cholesterol content of testis, the fructose content of the coagulating glands and the dorso-lateral prostate, the activities of alkaline phosphatase in the seminal vesicle, and acid phosphatase in ventral prostate of the MY treated animals suggested that their androgenic status were not affected. PMID- 9861483 TI - Bone density and mechanical properties in femoral bone of swim loaded aged mice. AB - Effects of swimming on bone density and mechanical properties of femur were investigated in aged male and female mice. R/1 strain of senescence accelerated mouse (SAM) at eleven months old was used. Two groups of males and two groups of females each consisting of 7 mice were used. One male and one female groups were loaded with a swim regiment of 40 min a day, 5 days a week for 6 consecutive weeks. The remaining groups were used as the controls. All mice were fed with the standard diet and water ad libitum during the experiments. The results of this study indicated that (i) the body weight was significantly (P < 0.05) lower in the swimming groups than in the control groups in both sexes. (ii) The bone density was significantly higher (P < 0.05) in the swimming groups than in the control groups in both sexes. However, there was no significant difference in cortical thickness index. (iii) In the mechanical properties of bone, there were no significant differences in the level of the maximum breaking force, the ultimate stress and the deformation between the swimming and the control groups in both sexes. However, the elasticity of the bone of the female mice in the swimming group was significantly higher (P < 0.05) than that of the control group. These results suggest that regimented swimming for the aged mice might suppress age-associated bone loss, and the effect of exercise in the females is greater that in the males. PMID- 9861484 TI - Mutagenicity of urine from individuals exposed to LPG combustion products. AB - The mutagenicity of urine from individuals exposed to the combustion products of liquefied petroleum gas (LPG) was detected with Salmonella typhimurium TA98 and its newly developed derivatives YG1021 (nitroreductase overproducing) and YG1024 (O-acetyltransferase overproducing). The detection showed significantly increased mutagenicity for the two YG strains and increased positive rates for all three strains in the presence of both rat liver S9 and beta-glucuronidase. Further analysis demonstrated that urine samples taken from smoking and non-smoking exposed individuals exhibited significantly higher mutagenic potency (revertants/10 microliters urine concentrate) than their corresponding controls. These results indicate that the increased urine mutagenicity is caused by the exposure to LPG combustion products or smoking. The mutagenic potency of urine samples of all exposed individuals tested with YG1024 was found to be about 7 times higher than with TA98. The difference in mutagenic potency was smaller for the same samples when comparison was made between YG1021 and TA98. This suggests that the mutagenic compounds present in the urine samples contain mainly aromatic compounds as glucuronide conjugates. Our results demonstrate that YG1024 is more sensitive than TA98 in detecting the mutagenicity of these samples. In addition, no significant difference in the mutagenic potency between the 'pure' exposed (non-smokers') and the 'pure' smokers' (unexposed) samples was found in all three tester strains. This might mean that the exposure extent of mutagens/carcinogens in LPG combustion products for exposed individuals roughly corresponds to the smoking level of smokers who smoke 20-40 cigarettes per day. Furthermore, the results also suggest that synergism might exist in the mutagenic effects of exposure to LPG combustion products and cigarette smoking. PMID- 9861485 TI - Antitumor activity of diallyl sulfide in two-stage mouse skin model of carcinogenesis. AB - It has been reported that diallyl sulfide (DAS), a sulfur-containing volatile compound in garlic (Allium sativum), exerts anticarcinogenic activity in various rodent tumor models. In the present study, the antitumor property of DAS was tested in Swiss albino mice in the two stage initiation-promotion mouse skin carcinogenesis. Skin cancers were initiated topically with a single subcarcinogenic dose (52 micrograms) of 7, 12-dimethyl benz (a) anthracene (DMBA). Promotion was performed by twice weekly applications of 12-O tetradecanoyl phorbol-13-acetate (TPA) at a dose of 5 micrograms/animal for 32 weeks. DAS was applied topically (250 micrograms/animal) thrice weekly for 3 weeks for anti-initiating and 1 h prior to each promotion treatment for anti promoting studies. The results showed that the treatment schedule of DAS can effectively delay the onset of tumorigenesis and reduce the cumulative number of tumors and the average number of tumors per mouse. In groups in which DAS applied prior to initiation or promotion, a significant population of the animals remained tumor-free till the termination of experiment. These findings suggest that DAS can effectively inhibit chemically induced mouse skin carcinogenesis. PMID- 9861486 TI - Child mortality patterns in rural areas of Anhui and Henan provinces in China, 1990. AB - County-based IMR and U5MR in Anhui and Henan provinces in China were estimated and analyzed by using the 1990 Census Data. Census was conducted on July 1, 1990, the number of deaths only occurred in the first half year of 1990 was collected. In order to obtain the total population and total number of deaths in the same year, the total number of deaths in each age-sex group for the whole 1990 was then estimated by taking the death number in the first half of 1990 as the base and multiplying a coefficient, which varied in different age-sex-region groups. Two major adjustments for some possible under-reporting cases in female birth and infant death were made. If the sex ratio at age 0 in some counties was beyond 1.2, then it was taken as 1.15 for rural counties and 1.10 for urban cities, which were the estimates of sex ratios for the children at age 5 in the national 1% Population Sampling Survey in 1995. The adjustment for IMR were made by comparing the segment of the county lift table from age 15 through 59 with that from the same age groups in the international and Chinese Model Life Tables. The IMR in the county life table would be substituted by the one in the closest Model Life Talbe, if it was less than in the latter. The findings of the analysis may be summarized as follows: (i) Total county-based IMR and U5MR were 33.4 per 1,000 and 41.4 per 1,000 respectively, with great variations between urban cities (25.4 per 1,000 for IMR and 31.4 per 1,000 for U5MR) and rural counties (35.1 per 1,000 for IMR and 43.6 per 1,000 for U5MR). There were also significant differences in child mortality between nationally identified poor counties and other counties in rural areas. In the poor counties the total IMR was 40.7 per 1,000 living births in average while in non-poor counties it was only 33.2 per 1,000 in average (P < 0.05). The U5MR in poor counties was 25 percent higher than in non-poor counties (51.5 vs 40.9 per 1,000 living births). (ii) Statistically significant correlation between child mortality and socio-economic variables was revealed from the data set, among which gross social economic products per capita was found to have the strongest relationship with child mortality. The negative correlation was found between child mortality and a set of so-called 'rich' variables including the gross social products, gross agricultural products, gross industrial products and the proportions of high-educated population at county level, whereas the positive correlation was found between child mortality and a set of 'poor' variables, such as proportions of residents with lower level of education and illiteracy rate. (iii) Differences in child mortality between these two provinces were found, which were identical to the trends of differences in socio-economic indicators between them. Lower child mortality proved to be associated with better socio-economic conditions (higher per capita products, higher proportions of residents with higher level of education, lower proportion of less educated people and illiteracy) in province Henan. (iv) A simple linear regression model was developed separately for Henan and Anhui to predict the IMR and U5MRs in each stage of economic development, where the dependent variables were the logarithm of IMR and U5MR, and the independent variables were the quintiles of the output value of gross products (GOP). It was found that at the first quintile, which was equivalent to 800 yuan of GOP in average, the predicted IMR and U5MR would reach 40 per 1,000 and 51 per 1,000 respectively. It would decline to 38 per 1,000 for IMR and 47 per 1,000 for U5MR in the second lowest quintile. Dramatic drop of child mortality was found between the second quintile and the third quintile, where 6 per 1,000 decline would occur for both IMR and U5MR. The decline would continue subsequently, but slower. The prediction of child mortality in rural counties could be used as a reference to assess counties at different stages of socio- PMID- 9861487 TI - A survey of social nutrition status of the elderly in the urban area of Chengdu, Sichuan Province. AB - The social nutrition status was investigated among 246 subjects aged 60-90 living at three urban communities in Chengdu of Sichuan Province. The questionnaire was designed to evaluate socio-demographic background, the subjects' nutrition knowledge, and the support systems for geriatric nutrition. Fasting venous blood was collected for the analysis of biochemical parameters. Blood pressure, bone mineral contents (BMC), body weight (BW) and body height (BH) were measured at the same time. Only 49.7% of the subjects correctly answered four basic questions on nutrition. Food patterns for the elderly were simple and modest. Several nutrition-related disorders for the elderly were including high systolic blood pressure (44.6%), hypertriglyceridemia (25.9%), high diastolic blood pressure (25.1%), obesity (24.5%), high PBG (20.6%), emaciation (19.9%), high FBG (17.9%) and osteoporosis (16.8%). These data indicate that the support systems for the geriatric nutrition will have to be improved. PMID- 9861488 TI - Resolution of enantiomers of ibuprofen by liquid chromatography: a review. AB - Ibuprofen is one of the most effective and widely used non-steroidal analgesic and anti-inflammatory agent. It is marketed as a racemic mixture though it is known that the pharmacological activity resides in the (S)-(+)-enantiomer only. Several direct/indirect liquid chromatographic methods involving a variety of chiral/achiral phases along with their possible role in resolution, chiral and achiral agents used for derivatisation have been discussed with special reference to ibuprofen, and mentioning their application to the resolution of other 2-aryl propionic acids/profens. PMID- 9861489 TI - Separatory determination of diastereomeric ibuprofen glucuronides in human urine by liquid chromatography/electrospray ionization-mass spectrometry. AB - A method for the separatory determination of diastereomeric isomers of glucuronic acid conjugates of ibuprofen having a carboxyl group at the chiral center by liquid chromatography (LC)/electrospray ionization (ESI)-mass spectrometry (MS) has been developed. The authentic specimens of acyl glucuronides of R(-)- and S(+)-ibuprofen were chemically synthesized by the Mitsunobu reaction. In the ESI mode, the glucuronides were characterized by an abundant quasi-molecular ion [M H]-, and the formation of the negative ion was markedly influenced by a drift voltage. The resolution of diastereomeric isomers was achieved on a Develosil ODS HG-5 column with 20 mM ammonium acetate (pH 5.0):acetonitrile (5:2, v/v) as a mobile phase where diastereomers were monitored with a corresponding quasi molecular ion. After oral administration of racemic ibuprofen, a preferential excretion of (S)-ibuprofen glucuronide into the urine was observed. PMID- 9861490 TI - Liquid chromatographic separation of some PTH-amino acids. AB - Liquid chromatographic studies on the separation of ten PTH-amino acids were carried out using normal phase untreated silica gel plates, C-18 RP precoated plates and RP-HPLC. Resolution of a complex mixture of PTH-amino acids was achieved using all the three types. Certain new successful solvent systems have been worked out in each case. HPLC was carried out with Lichrosphere 100 RP-18 (5 microns) column. Acetonitrile and sodium acetate buffer of pH 4.0 was used for reversed phase chromatography while for normal phase TLC combinations of chloroform-acetonitrile and chloroform-tetrahydrofuran were applied. PMID- 9861491 TI - A high-performance liquid chromatographic method for the determination of nicardipine in plasma and its application to pharmacokinetics in humans. AB - A simple and sensitive reversed-phase liquid chromatographic method has been developed and validated for the analysis of nicardipine in human plasma and the study of the pharmacokinetics of the drug in human body. Nicardipine and nimodipine (internal standard) in plasma were extracted with hexane-butanol (12:1, v/v) after addition of borate buffer (0.5 mol/mL, pH = 9.0), and then measured by HPLC using a Hypersil C18 column as stationary phase and acetonitrile -KH2PO4 buffer (0.015 M, pH = 5.5)--triethylamine as mobile phase. Nicardipine was quantified by ultraviolet absorbance at 236 nm. The method proved to be linear in the clinical range of 5-200 ng/mL with a regression coefficient of 0.9998. The lower limit of detection of nicardipine in plasma was 2.5 ng/mL. Intra- and inter-day coefficients of variation of assay for nicardipine in plasma were 3.5-5.4% (n = 7) and 5.2-6.4% (n = 5), respectively. The recovery of nicardipine was 92.8-100.8% for plasma. The method has been used to determine nicardipine in plasma samples from 10 volunteers and provided data on the pharmacokinetics of the drug. The results inferred that nicardipine is absorbed rapidly and has a relatively short half-life in healthy individuals. The data obtained was fitted with a 3P87 program to study the pharmacokinetics. The results showed that the disposition of nicardipine was conformed to a two compartment open model with Tmax = 1.6 +/- 0.3 h, Cmax = 109.8 +/- 38.7 ng/mL, T1/2 = 5.35 +/- 2.28 h and AUC0-->infinity = 322.1 +/- 69.6 ng/h/mI. PMID- 9861493 TI - Separation and determination of anthraquinone derivatives in rhubarb and its preparations by micellar electrokinetic capillary chromatography. AB - A micellar electrokinetic capillary chromatographic method was set up for the quality control of rhubarb and its preparations. Anthraquinone derivatives were separated successfully within 10 min in the buffer solution of 50 mmol/L H3BO3 NaOH (pH 11) containing 25 mmol/L sodium deoxycholate. The established method, with a recovery of extraction of over 90%, has good linear relationship and reproducibility. The contents of anthraquinone derivatives in rhubarb and a tablet of Niu-huang-jie-du differed significantly, showing that the quality control of rhubarb and its preparations is necessary. PMID- 9861492 TI - Continuous separation of serum proteins using a stirred cell charged with carboxylated and sulfonated microspheres. AB - We contrived a new separation system using a stirred cell charged with uncoupled microsphere similar to the chromatographic separation. Microspheres, carboxylated PS/PMAA and sulfonated PS/PNaSS, were prepared by emulsifier-free emulsion polymerization. To complement the submicron size weakness and the absence of ligands, we employed the latex form, the dispersion of microsphere, and took advantage of interaction relationships between proteins and microspheres. Adsorption isotherm is contemplated to investigate continuous separation behaviours of serum proteins. Selectivity of separation is in the following order: PS/PNaSS-2.0 (high sulfonated) < PS/PNaSS-0.3 (low sulfonated) < PS/PMAA 0.5 (low carboxylated). Unlike previous works on batch separation, not only the adsorbed amount in equilibrium (Cm), but also adsorption coefficient (K), played an important role in continuous separation. Functional groups (carboxyl and sulfonate), induced from the co-monomer, also affected the adsorption behaviours. PMID- 9861494 TI - Rapid determination of methotrexate in plasma by high-performance liquid chromatography with on-line solid-phase extraction and automated precolumn derivatization. AB - A high-performance liquid chromatographic system, combining solid-phase extraction and automated precolumn derivatization is described for the routine determination of methotrexate in human plasma. The sample extraction and elution onto the analytical column were performed automatically and concomitantly using the column-switching technique and a protein-coated precolumn. Cerium (IV) trihydroxyhydroperoxide (CTH) was introduced as a packed oxidant before the analytical column for the conversion of methotrexate into highly fluorescent products. The oxidative-cleavage of methotrexate occurs during the flow of 0.04 M phosphate buffer (pH 3.5) containing plasma sample through CTH column with a flow rate of 0.5 mL/min at 40 degrees C. The fluorescent products were transferred to the protein-coated precolumn, which was then flushed with the same buffer for clean-up and enrichment from plasma sample. The trapped substances were desorbed from the precolumn and eluted onto the ODS/TM analytical column by isocratical elution with a mobile phase containing 0.05 M phosphate buffer, pH 6.6 and acetonitrile (90-10, v/v) for subsequent separation. The fluorescent products were detected fluorimetrically at excitation and emission wavelengths of 367 and 463 nm, respectively. The complete analysis was achieved within 15 min per sample. The calibration graph was linear in the range of 50-500 ng/mL of methotrexate and there was no interference from endogenous components. PMID- 9861495 TI - Modulation of sialyl Lewis X dependent binding to E-selectin by glycoforms of alpha-1-acid glycoprotein expressed in rheumatoid arthritis. AB - Alpha-1-acid glycoprotein (AGP) is an extensively glycosylated acute phase protein of imprecisely defined physiological function. Nonetheless it is known that the oligosaccharide component comprising 42% of the 41 kDa molecular weight is critical to the previously described multifarious immunomodulatory functions of AGP in vitro. Complex oligosaccharides were enzymically released from AGP purified from the blood of rheumatoid arthritis sufferers by our oligosaccharide protective method. Oligosaccharide profiling was by means of high pH anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Monosaccharide composition analysis revealed increased fucosylation of inflammatory AGP oligosaccharide chains, suggesting the potential for expression of the tetrasaccharide antigen and E-Selectin ligand, sialyl Lewis X (sLeX). The hypothesis that AGP may function to inhibit blood cell binding to activated endothelium at E-Selectin was tested in a microtitre cell-protein binding assay. In this system we have shown that the oligosaccharide moiety of AGP, as expressed in inflammatory disease, can inhibit the sLeX/E-Selectin interaction. Thus we have identified a correlation between the abnormal glycosylation of AGP in rheumatoid arthritis and suppression of sLeX dependent cell adhesion through inhibition of E-selectin binding which could be the basis of a novel, site specific, anti-inflammatory agent. PMID- 9861497 TI - Ethicist misses opportunity to advocate pain control. PMID- 9861496 TI - Porphyrins in urine, plasma, erythrocytes, bile and faeces in a case of congenital erythropoietic porphyria (Gunther's disease) treated with blood transfusion and iron chelation: lack of benefit from oral charcoal. AB - Congenital erythropoietic porphyria is a rare genetic disorder in which deficiency of uroporphyrinogen III synthase results in excessive production of Type I porphyrins. The main clinical features are severe photodestruction of the skin and haemolytic anaemia. Treatment consists of shielding from light, blood transfusions and splenectomy, but is generally unsatisfactory. Previous studies have suggested that oral charcoal may be of benefit by binding porphyrins in the gut. A trial was therefore undertaken to evaluate this possibility. Porphyrins in urine, plasma and erythrocytes were measured by HPLC in a 23-year-old male patient with congenital erythropoietic porphyria, during an 8 week "run-in" period, and for a further 3 weeks when oral charcoal was given. Total urinary porphyrin excretion was 79-283 mumol/24 h consisting of 75% uroporphyrin I, 15% coproporphyrin I and smaller amounts of hepta-, hexa-, and pentacarboxylic porphyrins. Similar proportions were found in plasma and erythrocytes. During the first 24 h of charcoal administration a minor decrease in plasma and erythrocyte porphyrins was detected but this was not maintained during the remainder of the trial. In bile and faeces coproporphyrin I constituted approximately 95% of the porphyrins, with 2-3% coproporphyrin III and smaller amounts of pentaporphyrins I and III, but only trace amounts of uroporphyrin I. Oral charcoal was of no value in this case. Reasons are discussed in the context of biochemical differences between this patient with classical Gunther's disease and the similar clinical syndrome due to deficiency of uroporphyrinogen decarboxylase. PMID- 9861498 TI - Concern about the possibility of selenium in the human food chain. PMID- 9861499 TI - Efficacy of clindamycin hydrochloride capsules for the treatment of deep pyoderma due to Staphylococcus intermedius infection in dogs. AB - Clindamycin hydrochloride capsules (11 mg/kg body weight, q24 h) were administered orally to 20 dogs with deep staphylococcal pyoderma. Response to therapy was excellent in 100% of the dogs. Duration of therapy varied from 21 to 91 d, with an average duration of 45 d. Relapses occurred in 25% of the dogs within a 3-month period. One dog vomited when the clindamycin was given on an empty stomach. Under the conditions of the study, clindamycin was an effective, safe, and convenient antibiotic for the treatment of deep staphylococcal pyoderma in dogs. PMID- 9861500 TI - Cutaneous antimicrobial preparation prior to intravenous catheterization in healthy dogs: clinical, microbiological, and histopathological evaluation. AB - The purpose of this study was to determine the effects of a one-minute chlorhexidine gluconate skin preparation protocol prior to cephalic vein catheterization. Twenty-three healthy beagle dogs had one leg aseptically prepared and the opposite leg served as a control. Twenty-six- and 77-hour time groups were studied. Chlorhexidine-treated legs had significantly lower cutaneous bacterial counts than the control legs prior to catheter insertion and prior to catheter withdrawal for both time groups. Control legs developed significantly more dermatitis than the treated legs after 77 h. A one-minute preparation with 4% chlorhexidine gluconate was an effective method for sustained reduction of cutaneous bacterial counts at peripheral intravenous catheter insertion points in dogs. Increased cutaneous bacterial counts were associated with significantly more microscopic dermatitis in untreated legs after 77 h of catheterization. PMID- 9861501 TI - Postcastration eventration in 18 horses: the prognostic indicators for long-term survival (1985-1995). AB - Castration of horses is considered a common and routine surgical procedure, but the potential for complications is high. By far the most serious of these is eventration. The objectives of this study were to determine the long-term survival of horses undergoing surgical treatment of indirect (1) inguinal eventration of the small intestine following castration, and to identify prognostic indicators for survival. The case records of 18 horses undergoing surgical treatment of postcastration eventration (PCE) between 1985 and 1995 were reviewed. Follow-up information was obtained by telephone interviews 2 to 13 y postoperatively. A Cox proportional hazard regression model was fitted to determine which clinical features were of significant influence to survival. Clinical features with a significant negative influence on survival were an inguinal surgical approach for correction, an increased length of prolapsed bowel, and performance of bowel resection and anastomosis. Significant postoperative complications developed in 89% of cases; 44% of cases in the "inguinal" surgical approach group developed peritonitis, compared with 10% in the "midline" approach group. Of all horses in this study, 72% were discharged from the hospital; however, only 40% of horses in the inguinal approach group were discharged. The long term survival rate (> 1 y) for all horses in this study was 44%, with a median survival time of 3-1/2 mo. PMID- 9861502 TI - Partial intestinal obstruction due to colonic adenocarcinoma in a cat. AB - An abdominal mass was palpated in a 14-year-old, spayed female cat with unresolving diarrhea and decreased appetite. A stricture of the ascending colon was confirmed radiographically, identified upon laparotomy, and resected. Colonic adenocarcinoma with infiltration into lymphatics was diagnosed histologically. Eight months postoperatively, the subject was clinically normal. PMID- 9861503 TI - Canada's subnational surveillance roots. PMID- 9861504 TI - Added value to veterinary service. PMID- 9861505 TI - Evolution of web site design: implications for medical education on the Internet. AB - Since its inception, the world wide web (WWW) has possessed the potential for becoming a 'watershed' medium for conveying complex, structured information across vast temporal and geographical barriers. In 1995, the MedWorld project (http:(/)/medworld.stanford.edu) was created at the Stanford University School of Medicine in an effort to innovate and explore the design process of creating WWW applications specifically for medical education. Until recently, the evolution of WWW applications has been mainly driven by technological advances in client server technology, enabling or translating traditional modes of collaborative medical education (e.g. voice, presence, print, motion) into WWW devices and applications. Many of these applications, while technologically advanced, lack focused development of interface and interactivity design, which may enhance learning experiences. WWW applications which incorporate design innovation in parity with advances in client-server technology have been termed, 'third generation' web sites and have the potential to improve the quality of WWW applications designed for medical education. This work describes how the MedWorld project has created a 'third generation' WWW application by utilizing innovation in information, interface and interactivity design to create innovative WWW technology for the medical education arena. PMID- 9861506 TI - IDEM: a Web application of case-based reasoning in histopathology. AB - Different software engineering and artificial intelligence methods can be used to design Internet retrieval of prototypical medical images. We used the case-based reasoning (CBR) approach to provide an 'intelligent' access to a collection of illustrated medical cases through the Internet. This paper presents a Web interface for the CBR system IDEM (image and diagnosis from examples in medicine) in the domain of breast pathology. Thanks to the definition of a similarity measure between the descriptions of cases we propose a flexible querying of the case-base and a quantitative browsing among cases through similarity links. The resemblance rates provided by the system argue for the quality and the relevancy of the retrieved data. The flexibility of the querying process is robust to missing information and could be adapted to a daily practice. The CBR approach is a promising method for a clinical relevant and an efficient retrieval of reference images and diagnosis clues through Internet. PMID- 9861507 TI - Computer network home care demonstration: a randomized trial in persons living with AIDS. AB - PURPOSE: As Internet-based health care applications grow, it is important to determine their acceptability to ill persons and the likely outcomes resulting from their use. This randomized field investigation demonstrated the use and effects of a specialized computer network, the ComputerLink, among persons living with AIDS (PLWA). HYPOTHESIS: This study was designed to determine whether a home based computer network designed for people with AIDS (PLWA) would be used and whether it would reduce social isolation and improve confidence and skill in decision making without causing differential decline in health status among PLWA. INTERVENTION: Wyse 30 terminals were placed in private homes, linked via a 1200 baud modem to a public access computer network. Services available to PLWA included an on-line electronic encyclopedia, public and private communication and a decision support system; all services were coordinated by a registered nurse. DESIGN: A six-month randomized trial involved 57 community-dwelling PLWA; 31 were in the experimental group. Mean age was 33, 93% were male, 61% white, 34% working, 13.5 mean years education. Six participants were lost to follow-up. RESULTS: 8449 accesses to the system were made during the study period. Mean number of access per participant was 192, the median, 129. Duration of access averaged 12 min. PLWA used two services during each session. First order hypotheses testing revealed no significant difference between experimental and control group participants. Post hoc exploration indicated that use of the system did reduce social isolation once participants levels of depression were controlled and that decision making confidence improved as a function of number of accesses. CONCLUSION: Computer networks provide feasible alternatives for the delivery of health services to home-bound individuals. Communication services were used more extensively than other services, suggesting that the primary mechanism of intervention is peer contact. Similar to other experiments, system use improves confidence, though not skill, in decision making. The Internet represents a promising pathway to reaching home-bound patients and providing them with information, communication and decision assistance. PMID- 9861508 TI - The telemedicine information exchange: an online resource. AB - Telemedicine is the use of telecommunications to provide health care services at a distance. It has grown from mostly government-subsidized research initiatives into a fledgling industry. This growth was fueled by decreasing costs of telecommunications and information technologies and fanned by rising health care costs. The telemedicine information exchange (TIE) (http:(/)/tie.telemed.org), developed by the Telemedicine Research Center in Portland, OR, is both a product and a vehicle of this rapid growth. It facilitates collection of telemedicine information from many sources, providing an easy-to-use hypertext format. This article describes the TIE's development, advantages and disadvantages of a web based online library, and it's codevelopment with a rapidly expanding industry. PMID- 9861509 TI - Internet and healthcare in Brazil: the role of the Working Group for Healthcare (GT Saude). AB - This paper introduces the GT Saude initiative in Brazil. The group is part of the Brazilian Internet project, which goals are to depict how Internet can be useful to the society. The mission of the GT Saude group is to foster Internet applications on the healthcare area. The main projects currently under development are presented: Multicom-21, using large bandwidth connections on telemedicine; the Unified Health Record, using Internet and smart-cards for a national minimum patient data set; the Brazilian Virtual Hospital, that presents a large amount of useful information and links for healthcare professionals and people in general; the National Healthcare Information Network (RNIS), that uses Internet as a medium for data exchange among the state secretaries and publishing statistics from the national healthcare system (SUS) for general public access; and finally the virtual university proposal, which uses Internet for education and is offering its first course on nutrition. The heterogeneity of the projects is meant and is part of the group's task, which is to cover the subject as widely as possible. Through this 'demonstration' projects the group is trying to prove the usefulness and benefits of using Internet technologies, even (or mainly) on a developing country with an inadequate healthcare situation. In its short existence the GT Saude group has achieved several of its original goals. The most relevant accomplishments are: putting together different groups, eliciting synergy across the projects and encompassing a broad spectrum of applications (the 'demonstration factor'). PMID- 9861510 TI - WWW-based access to object-oriented clinical databases: the KHOSPAD project. AB - KHOSPAD is a project aiming at improving the quality of the process of patient care concerning general practitioner-patient-hospital relationships, using current information and networking technologies. The studied application field is a cardiology division, with hemodynamic laboratory and the population of PTCA patients. Data related to PTCA patients are managed by ARCADIA, an object oriented database management system developed for the considered clinical setting. We defined a remotely accessible view of ARCADIA medical record, suitable for general practitioners (GPs) caring patients after PTCA, during the follow-up period. Using a PC, a modem and Internet, an authorized GP can consult remotely the medical records of his PTCA patients. Main features of the application are related to the management and display of complex data, specifically characterized by multimedia and temporal features, based on an object-oriented temporal data model. PMID- 9861511 TI - Modeling and implementing a database on drugs into a hospital intranet. AB - Our objective was to develop a drug information service, implementing a database on drugs in our university hospitals information system. Theriaque is a database, maintained by a group of pharmacists and physicians, on all the drugs available in France. Before its implementation we modeled its content (chemical classes, active components, excipients, indications, contra-indications, side effects, and so on) according to an object-oriented method. Then we designed HTML pages whose appearance translates the structure of classes of objects of the model. Fields in pages are dynamically fulfilled by the results of queries to a relational database in which information on drugs is stored. This allowed a fast implementation and did not imply to port a client application on the thousands of workstations over the network. The interface provides end-users with an easy-to use and natural way to access information related to drugs in an internet environment. PMID- 9861512 TI - Internet integrated in the daily medical practice within an electronic patient record. AB - Healthcare enters the information age and professionals are finding an ever growing role for computers in the daily practice of medicine. However, a number of problematic issues are associated with electronic publications, especially through Internet. Whilst access to any information has been improved, access to specific information has become more and more difficult [1], due to the lack of a general meta-knowledge allowing to structure Internet resources. Physicians have to learn and adapt themselves to computers and Internet, but Internet has to meet the specific requirements of Healthcare. Important issues must therefore be addressed to allow a real and daily use of Internet in the medical practice. The paper discusses most of these issues and proposes a solution developed at the University Hospital of Geneva that integrates an Electronic Patient Record with Internet, without compromises on security or on performances and that runs on standard PCs'. PMID- 9861513 TI - Usage analysis of a primary care medical resource on the Internet. AB - Physicians and patients need convenient access to quality medical information. This study's goal was to place a medical resource on the World-Wide Web (WWW), allow access to it through a simple to use interface, and analyze the usage of such a resource. The Family Practice Handbook (TFPH) was digitized and placed onto the WWW. Usage data was obtained from June 1995-June 1996. 118,804 individuals accessed TFPH viewing 409,711 pages of information. A broad spectrum of topics was accessed. TFPH proved to be an extremely popular resource, servicing the broad information needs of an international audience. These preliminary findings suggest the future promise of Internet medical resources. PMID- 9861514 TI - Trends in medical information retrieval on Internet. AB - Information on the World Wide Web is unstructured, distributed, multimedia and multilingual. Many tools have been developed to help users search for useful information: subject hierarchies, general search engines, browsers and search assistants. Although helpful, they present serious limitations, mainly in terms of precision, multilingual indexing and distribution. In this paper, we cover some on-line solutions to medical information discovery and present our own approach, the MARVIN (multi-agent retrieval vagabond on information network) project, which tackles medical information research with specialized cooperative retrieval agents. We also draw some outlines for future extensions. PMID- 9861515 TI - The Health On the Net Code of Conduct for medical and health Websites. AB - Internet has become one of the most used communication media. This and the fact that no constraining information publishing policy exists have created an urgent need to control the quality of information circulating through this media. To this purpose, the Health On the Net Foundation has initiated the Code of Conduct (HONcode) for the health/medical domain. This initiative proposes guidelines to information providers, with the aim, on the one hand, of raising the quality of data available on the Net and, on the other hand, of helping to identify Internet sites that are maintained by qualified people and contain reliable data. The HONcode mainly includes the following ethical aspects: the author's credentials, the date of the last modification with respect to clinical documents, confidentiality of data, source data reference, funding and the advertising policy. This article presents the HONcode and its evolution since it was launched in 1996. PMID- 9861516 TI - Deposition of oxidized low-density lipoprotein and collagenosis occur coincidentally in human coronary stenosis: an immunohistochemical study of atherectomy. AB - BACKGROUND: Coronary stenosis involves lipid accumulation, fibrosis and cell proliferation. OBJECTIVE: To clarify the role of oxidized low-density lipoprotein (LDL) in coronary stenosis by examining atherectomized coronary lesions from patients with primary stenosis and restenosis after percutaneous transluminal coronary angioplasty (PTCA). METHODS: Atherectomized coronary tissue from 28 patients with primary stenosis and restenosis at 4.3 +/- 1.0 months after PTCA were examined using morphometrical and immunohistochemical techniques. RESULTS: Serum lipids in all of the patients were within the normal range and no differences were noted between the two groups. There were no differences in the mean cross-sectional areas of whole specimens obtained from each group, and sclerotic lesions with atheroma or calcification were found to a similar extent in both groups. However, the restenosis group had a significantly greater area (6 fold) of immature smooth-muscle-rich lesions than the primary stenosis group, although there was no difference in lipid-laden foam-cell containing lesions. In foam-cell-containing lesions, apolipoprotein B was accumulated extracellularly, while oxidized LDL was primarily deposited intracellularly in lipid-laden foam cells. However, no deposition of apolipoprotein B, oxidized LDL or lipids was noted in smooth-muscle-rich lesions. Proline hydroxylase, a key enzyme for collagen synthesis, was detected in most of the foam-cell-containing lesions, but not in smooth-muscle-rich lesions. CONCLUSIONS: Atherectomized lesions from patients with coronary stenosis contained smooth-muscle-rich lesions in restenosis and lipid-laden cellular lesions in both stenosis and restenosis, in which the deposition of oxidized LDL and increased collagen synthesis occur coincidentally. Therefore, the mechanism of atherogenesis may involve coronary stenosis regardless of the occurrence of restenosis after PTCA therapy. PMID- 9861517 TI - Association of central obesity and insulin resistance with high prevalence of diabetes and cardiovascular disease in an elderly population with low fat intake and lower than normal prevalence of obesity: the Indian paradox. AB - OBJECTIVE: To test the hypothesis that high prevalences of cardiovascular disease and diabetes in urban population of India are manifestations of insulin resistance syndrome. DESIGN AND SETTING: Cross-sectional surveys were conducted in 20 streets in the city of Moradabad and in two villages. SUBJECTS AND METHODS: There were 566 subjects (255 rural and 311 urban) aged 60-84 years. All subjects were divided on the basis of their insensitivity to insulin into groups with mild, moderate and high insensitivity to insulin and data from both sexes were pooled for analysis. RESULTS: Overall increases in the prevalences of coronary disease, diabetes, hypertension, central obesity and associated disturbances were observed with increasing insensitivity to insulin and the trend was more significant among urban than it was among rural subjects. Multivariate logistic regression analysis revealed a significant positive association of level of insulin insensitivity with the age-adjusted prevalences of coronary disease, hypertension, diabetes, hypertriglyceridaemia, intolerance of glucose and central obesity among urban subjects. We observed a significant inverse association between insensitivity to insulin and physical activity both for rural and for urban subjects and between insensitivity to insulin and high-density lipoprotein cholesterol level for urban subjects. For rural subjects, we found significant associations of sensitivity to insulin with coronary artery disease and intolerance of glucose without significant associations with other risk factors. CONCLUSION: Insensitivity to insulin was significantly associated with risks of cardiovascular disease and diabetes, despite there being a low prevalence of obesity (9.0%) among urban subjects. Hypertension, diabetes, hypertriglyceridaemia, intolerance of glucose and central obesity were significantly associated with insensitivity to insulin and coronary disease for urban but not for rural people. PMID- 9861518 TI - Assessment of left ventricular wall motion and delineation of the endocardial border after intravenous injection of Infoson during dobutamine stress echocardiography. AB - OBJECTIVES: To investigate whether intravenous injection of Infoson facilitates the assessment of left ventricular wall motion and definition of endocardial border and thereby reduces inter- and intra-observer variability during dobutamine stress echocardiography. BACKGROUND: Clear detection of the endocardial border is essential during dobutamine stress echocardiography. Although several contrast agents have been tested for their efficacy in enhancing definition of left ventricular endocardium, their usefulness during dobutamine stress echocardiography has not been evaluated. METHODS: Thirty coronary artery disease patients underwent dobutamine stress echocardiography. Infoson was injected at 0.2 ml/kg in both apical four- and two-chamber views. Detection of the left ventricular endocardial borders was scored from 0 (undetectable) to 10 (best) and expressed as a percentage of image quality at rest and peak stress by two independent observers. Regional wall motion was also evaluated and the total wall motion score index calculated each time. RESULTS: Delineation of the left ventricular endocardium improved from 76 +/- 4% to 84 +/- 2% at rest (P < 0.01) and from 75 +/- 4% to 89 +/- 1% at peak stress (P < 0.01) after administration of Infoson. The greatest improvement was seen in the basal and middle regions of the lateral and anterior walls. Inter- and intra-observer variability was reduced after administration of Infoson. At rest, the probability of concordance between two observers increased from 0.86 (0.82-0.89) to 0.91 (0.88-0.94) (P < 0.05) and at peak stress from 0.86 (0.82-0.9) to 0.90 (0.86-0.92) after administration of Infoson (P < 0.05). The probability of concordance between on- and off-line assessment by one observer also increased from 0.84 (0.8-0.88) to 0.90 (0.86 0.93) (P < 0.01) at rest and from 0.90 to 0.92 (NS) at peak stress. Overall, a change in wall motion score index occurred in 16 of 30 (53%) patients after administration of Infoson, thus improving the accuracy of the stress test compared with coronary angiography. The wall motion score index was overestimated in 11 of 16 (37%) patients without Infoson while the detection of new wall motion abnormalities increased in 5 of 16 (17%) (P < 0.01). CONCLUSIONS: Intravenous administration of Infoson facilitates the assessment of wall motion, particularly of the basal lateral and anterior walls where endocardial border drop-out frequently occurs during dobutamine stress echocardiography; it thus reduces the inter- and intra-observer variability. PMID- 9861519 TI - Coronary artery stent placement with postprocedural antiplatelet therapy in acute myocardial infarction. AB - BACKGROUND: We have shown that coronary artery stent implantation in acute myocardial infarction (AMI) is feasible and safe when combined with effective postprocedural antiplatelet therapy. However, the concept of coronary stenting in AMI has not been validated in large cohorts. In this observational study, we investigated the outcome in 318 consecutive patients. METHODS AND RESULTS: From January 1995 until December 1996, 420 of 455 (92.3%) patients admitted to our institutions with AMI underwent infarct artery stent placement. This report describes the 318 patients treated with combined antiplatelet therapy whose AMI was not complicated by cardiogenic shock or mechanical ventilation before the intervention. Postinterventional therapy consisted of 100 mg aspirin and 250 mg ticlopidine twice daily. There were 21 (6.6%) cardiac events during 30-day follow up: five cardiac deaths (1.6%), four nonfatal re-infarctions (1.3%) and 13 target vessel revascularizations (4.1%). Six months of clinical follow-up yielded a 94.7% survival rate free of repeat AMI. The rate of repeat interventions was 11%. Quantitative computerized angiography at 6 months, performed in 79.4% of the eligible patients, revealed a binary restenosis rate of 25%. CONCLUSIONS: With postprocedural antiplatelet therapy, coronary stenting in AMI yields favorable short- and long-term outcomes. PMID- 9861520 TI - Elderly patients with hypercholesterolaemia: a double-blind study of the efficacy, safety and tolerability of fluvastatin. AB - BACKGROUND: Coronary heart disease is a major cause of morbidity and mortality in the elderly, a rapidly growing section of the population. Elderly patients have been excluded from most preventative risk factor trials. METHODS: We evaluated fluvastatin, a fully synthetic hydroxymethyl glutaryl coenzyme A reductase inhibitor, in white patients older than 60 years, in seven hospital centres. After an 8-week cholesterol-decreasing diet phase, patients were allocated to groups to receive fluvastatin 40 mg daily (n = 33) or placebo (n = 36) given for 12 weeks. All patients had low-density lipoprotein cholesterol concentrations > or = 4.1 mmol/l 1 week before they were allocated to a treatment at random. After receiving randomised treatment for 12 weeks, 50 patients then received fluvastatin 40 mg daily on an open basis for a further 12 weeks. RESULTS: Mean +/ SD age was 70.7 +/- 5.2 years for fluvastatin patients and 68.3 +/- 5.6 years for placebo. Mean +/- SD percentage changes in lipid concentrations from randomisation to the end of 12 weeks were calculated (n = 63) by intent-to-treat analysis. Total cholesterol decreased by 21.64 +/- 8.7% in the fluvastatin group and by 2.91 +/- 7.25% in the placebo group (P < 0.01); high-density lipoprotein cholesterol increased by 4.98 +/- 10.84% in the fluvastatin group and decreased by 0.05 +/- 8.68% in the placebo group (P = 0.05); low-density lipoprotein cholesterol decreased by 27.14 +/- 8.45% in the fluvastatin group and by 2.16 +/- 9.68% in the placebo group (P < 0.01); very-low-density lipoprotein cholesterol decreased by 30.70 +/- 30.65% in the fluvastatin group and by 9.80 +/- 28.6% in the placebo group (P < 0.01); triglyceride decreased by 18.13 +/- 17.35% in the fluvastatin group and by 2.97 +/- 21.85% in the placebo group (P < 0.01). There were no statistically significant differences between treatment groups for any other biochemical or haematological parameters. Adverse events were mainly mild, diminishing with continued treatment, and no event was serious by standard criteria. Patient-assessed tolerability after randomised treatment was 'very good' for 18 fluvastatin patients and for 26 placebo patients (P = 0.79). Seven patients withdrew from the 12-week follow-up (four from the fluvastatin group and three from the placebo group). CONCLUSIONS: We conclude that fluvastatin decreases lipid concentrations effectively and safely in elderly patients, producing clinically significant decreases in total cholesterol, low-density lipoprotein cholesterol, triglyceride and, especially, very-low-density lipoprotein cholesterol, while increasing high-density lipoprotein cholesterol moderately. PMID- 9861521 TI - Efficacy and safety of adenosine in diagnosis and treatment of regular tachycardia in the elderly. AB - BACKGROUND: Adenosine is currently used in the treatment and differential diagnosis of regular tachycardia. However, the efficacy of its employment has not been studied in elderly people. METHODS: We evaluated the safety, and the diagnostic and therapeutic utility of adenosine in elderly people aged over 70 years with regular sustained tachycardia, compared with a group of patients aged under 70 years affected by the same arrhythmia. Adenosine was given to 107 patients in increasing bolus doses up to 18 mg during regular broad and narrow complex tachycardia; 49 patients were aged 70 and over, and 58 patients were aged less than 70 years. In the former group, surface ECG showed 38 narrow complex tachycardias and 11 broad complex ones; in the second group there were 48 narrow complex tachycardias and 10 broad complex ones. RESULTS: Adenosine was effective in 94% of the elderly patients and in 93% of the younger patients. In the group aged over 70 years, adenosine restored sinus rhythm in 37% of patients and revealed the mechanism of arrhythmia in 57%. Adenosine restored sinus rhythm in 50% of patients under 70 years and revealed atrial or sinus tachycardia in 43%. The incidence of symptomatic side effects and peri-conversion ventricular arrhythmias was similar in the two groups. There were ventricular pauses over 3 s long in four (8%) of the older patients (maximum pause 7 s) and in two patients (3%) of the group under 70 years (maximum pause 6 s). No adverse haemodynamic effects were observed. CONCLUSION: These data demonstrate the safety and the value of adenosine in the diagnosis and treatment of regular tachycardia in elderly patients. PMID- 9861522 TI - Hormonal response to orthostasis in elderly people with systemic systolic hypertension. AB - BACKGROUND: Orthostatic hypotension is a common phenomenon in the elderly. Hormonal changes during orthostatic stress have been described in elderly normotensive people and in those with essential hypertension. However, the hormonal response in elderly people who have systolic hypertension during orthostasis has not yet been quantified. METHODS: In this study we investigated 14 non-diabetic men, aged 60 to 75 years, with untreated systolic hypertension who were subjected to 45 degrees passive head-up incline on a tilt table for 15 min. Their hormonal profile and hemodynamic changes were analyzed before and after the stress. RESULTS: In the supine position, plasma levels of norepinephrine, atrial natriuretic peptide and aldosterone were in the normal range, while the plasma renin activity was low. Immediately upon tilt the systolic blood pressure fell but it reverted to baseline values after 15 min of orthostasis. At that time the cardiac output decreased while the systemic vascular resistance and the plasma norepinephrine concentration rose. The atrial natriuretic peptide appeared to fall, and the renin-aldosterone level did not change. CONCLUSION: The physiologic response to orthostatic stress in elderly people with systolic hypertension is comparable to that of elderly normotensive people and those with essential hypertension, i.e. a decrease in cardiac output and an increase in plasma norepinephrine levels. The atrial natriuretic peptide appeared to fall appropriately. The response of the renin-aldosterone system mimicked that in elderly patients with low renin essential isolated hypertension. These observations may have a bearing on the management of elderly people with systolic hypertension who also have orthostatic symptoms; they may not require a different approach from that needed for others of the same age group. PMID- 9861524 TI - Bibliography current world literature. PMID- 9861523 TI - Pharmacology of the intravenous platelet receptor glycoprotein IIb-IIIa antagonists. PMID- 9861525 TI - Offspring sex ratio in the assessment of the effects of endocrine modulators. PMID- 9861526 TI - Dioxin-like and non-dioxin-like toxic effects of polychlorinated biphenyls (PCBs): implications for risk assessment. AB - Polychlorinated biphenyls (PCBs) are persistent, bioaccumulative, and toxic contaminants in the environment. Individual PCB congeners exhibit different physicochemical properties and biological activities that result in different environmental distributions and toxicity profiles. The variable composition of PCB residues in environmental matrices and their different mechanisms of toxicity complicate the development of scientifically based regulations for the risk assessment. In this article various approaches for the assessment of risks of PCBs have been critically examined. Recent developments in the toxic equivalency factor (TEF) approach for the assessment of toxic effects due to dioxin-like PCBs have been examined. PCB exposure studies that describe non-dioxin-like toxic effects, particularly neurobehavioral effects and their effective doses in animals were compiled. A comparative assessment of effective doses for dioxin like and non-dioxin-like effects by PCBs has been made to evaluate the relative significance of non-ortho-and ortho-substituted PCBs in risk assessment. Using mink as an example, relative merits and implications of using TEF and total PCB approaches for assessing the potential for toxic effects in wildlife was examined. There are several advantages and limitations associated with each method used for PCB risk assessment. Toxic effects due to coplanar PCBs occur at relatively smaller concentrations than those due to non-dioxin-like PCBs and therefore the TEF approach derives the risk assessment of PCBs, in the environment. The need for the refinement of TEF approach for more accurate assessment of risks is discussed. PMID- 9861527 TI - Toxicokinetics of organic solvents: a review of modifying factors. AB - This article reviews, with an emphasis on human experimental data, factors known or suspected to cause changes in the toxicokinetics of organic solvents. Such changes in the toxicokinetic pattern alters the relation between external exposure and target dose and thus may explain some of the observed individual variability in susceptibility to toxic effects. Factors shown to modify the uptake, distribution, biotransformation, or excretion of solvent include physical activity (work load), body composition, age, sex, genetic polymorphism of the biotransformation, ethnicity, diet, smoking, drug treatment, and coexposure to ethanol and other solvents. A better understanding of modifying factors is needed for several reasons. First, it may help in identifying important potential confounders and eliminating negligible ones. Second, the risk assessment process may be improved if different sources of variability between external exposures and target doses can be quantitatively assessed. Third, biological exposure monitoring may be also improved for the same reason. PMID- 9861528 TI - Evaluation of automated systems for the analysis of cervical smears. PMID- 9861529 TI - The accuracy of endometrial cytology in the diagnosis of endometrial adenocarcinoma. AB - We have examined 62,234 cytological samples of endometrium to establish the accuracy and false-positive rate for the diagnosis of endometrial adenocarcinoma. The patients were either attending the gynaecological out-patients clinic with symptoms or were asymptomatic women attending for routine population screening as part of our cancer detection programme, the numbers from these two sources being equal. Out of 138 cases identified as endometrial adenocarcinoma by cytology 126 (91.3%) were confirmed histologically in our hospital. Twelve cases (8.7%) were shown to be false-positives. Reexamination of these led to the same false positive diagnosis in all 12 cases. This was attributable to similarities of nucleo-cytoplasmic ratio, irregular arrangement of nuclei, variation in nuclear shape and in the numbers of nucleoli in repair cells and hyperplastic cells compared with the carcinoma cases. Most of the false-negative reports were due to insufficient material, pale staining in malignant cells or diagnostic error. Refraction measurement of the density of nuclei of cancer cells using equipment for which the patent is pending enabled objective measurement of nuclear density which indicated that the nuclei were not stained darkly enough in false-negative cases. PMID- 9861530 TI - Aspergillus in the Papanicolaou stain: morphology, fluorescence and diagnostic feasibility. AB - Aspergillus species exhibit a distinct and clear fluorescence in Papanicolaou stained cytological samples. The Papanicolaou (PAP) stain enhances the autofluorescence of cultured aspergilli and allows better cytological recognition of the fungus by fluorescence microscopy when it is not easily discerned from its surroundings by light microscopy. Morphological properties can be better distinguished and facilitate the differentiation of aspergillus organisms from other filamentous fungi. Neither light nor fluorescence microscopy, the cytological quality nor the presence of phagocytosed hyphae in alveolar macrophages allow distinction between infection and contamination with Aspergillus species. Only the presence of eosinophilic inflammation permits a tentative diagnosis of an Aspergillus infection. In conclusion, PAP fluorescence reduces the need for special stains, is superior to and quicker than other investigative techniques and enhances the sensitivity and specificity of cytological investigation when a rapid and reliable identification of Aspergillus is needed. PMID- 9861531 TI - Cytologic diagnosis and subtyping of rhabdomyosarcoma. AB - We reviewed the cytological findings of 38 cases of rhabdomyosarcoma (RMS) with histological confirmation performed during a period of 15 years and proposed a morphological subtyping based on the most prominent cytologic features. Seventeen of these cases were alveolar, 14 cases embryonal, and seven botryoid subtypes. From these cases, a total of 43 samples, of which 37 were fine needle aspiration (FNA) biopsies and six were touch imprints, were evaluated. Detailed cellular features were identified which enabled differentiation into histological subtypes. In the alveolar RMSs, most tumour cells were small and lymphocyte-like, having finely granular chromatin. The finding of cells with more abundant cytoplasm, eccentrically located nuclei and bi/multinucleated tumour cells in a background of mucosubstance helped in the differential diagnosis. Two cell types, including large, tadpole or ribbon-shaped tumour cells and small, round cells with scant cytoplasm, were seen in embryonal RMSs. In botryoid RMSs, a cell type with tightly grouped nuclei within elongated cytoplasm similar to a myotubular structure was observed in addition to the two cell types of embryonal RMSs. We conclude that with experience it will be possible to subtype these tumours by cytologic examination alone. PMID- 9861532 TI - The use of the PAPNET automated cytological screening system for the diagnosis of oral squamous carcinoma. AB - The automated PAPNET screening system has been developed to recognize abnormal cells in cervical smears. Given that the oral mucosa sheds cells resembling superficial and intermediate cells of the cervix, the aim of this study was to assess whether the PAPNET system could be used to detect dysplastic cells in oral mucosal smears. Sixty-two oral smears from 27 patients were examined by both light microscopy and using the PAPNET system from clinically abnormal and normal areas by two pathologists. The clinically abnormal sites were also biopsied for histological analysis. There was 100% correlation between the manual and PAPNET screening results. Cytological interpretation of oral smears by both manual and PAPNET screening methods correctly diagnosed squamous cell carcinoma in 14/23 (61%) of patients who had all been confirmed by biopsy. The nine patients with false-negative cases could be attributed to poor smear technique and preparation. The PAPNET system can be used to identify abnormal cells in oral smears and, as such, may have an application for screening those populations at high risk of oral cancer--provided that adequate tuition is given in smear technique. PMID- 9861533 TI - Use of bile cytology for early diagnosis of complications in orthotopic liver transplantation. AB - We conducted a daily analysis of bile cellularity in 25 patients undergoing 29 orthotopic liver transplants (OLT) and correlated the cytological parameters with the clinical outcome of each patient. The 16 patients without complications only showed slides with cells during the first 4-5 postoperative days. The four patients with primary non-function (PNF) of the graft had a high cell density up to the time of the retransplant, with a preponderance of polymorphonuclear (PMN) leucocytes (59.2%) and epithelial cells (29.2%). During the episodes of sepsis (n = 3) and rejection (n = 7) we noted the sudden appearance of high cellularity, almost exclusively PMN leucocytes (96.5%), and a preponderance of PMN leucocytes (84.2%) with appreciable percentages of mononuclear cells and macrophages as well as the early appearance of lymphoblasts in the rejection episodes. Our results show that bile cytology can be a useful method for diagnosing graft complications in liver transplantation. PMID- 9861534 TI - Fine needle aspiration (FNA) cytology of primary subcutaneous sacrococcygeal myxopapillary ependymoma. PMID- 9861535 TI - False-negative cervical smears. PMID- 9861536 TI - False-negative cervical smears. PMID- 9861537 TI - Cover slip sizes for routine cervical screening. PMID- 9861538 TI - Leptin and its receptors: regulators of whole-body energy homeostasis. AB - Leptin is the adipocyte-specific product of the ob gene. Expression of leptin in fully fed animals reflects adipocyte size and body-fat mass. Leptin signals the status of body energy stores to the brain, where signals emanate to regulate food intake and whole-body energy expenditure. The leptin gene was identified in the leptin-deficient, obese ob/ob mouse by positional cloning techniques. Recently, leptin has been cloned in domestic species including pigs, cattle, and chickens. The leptin receptor has at least five splice variants; the long form of the receptor is primarily expressed in the hypothalamus and is thought to be the predominant signaling isoform. Leptin receptors are members of the cytokine family of receptors and signal via janus-activated kinases (JAK)/signal transducers and activators of transcription (STAT) and mitogen-activated protein kinase (MAPK) pathways. Mutations in the leptin or leptin receptor genes results in morbid obesity, infertility, and insulin resistance in rodents and humans. Leptin regulates food intake and energy expenditure via central and peripheral mechanisms. Leptin receptors are expressed in most tissues, and in vitro evidence suggests that leptin may have direct effects on some tissues such as adipose tissue, the adrenal cortex, and the pancreatic beta-cell. Leptin is thought to influence whole-body glucose homeostasis and insulin action. Studies are underway to determine the role that leptin plays in the biology of domestic animals. PMID- 9861539 TI - Cellular mechanisms by which oxytocin mediates uterine prostaglandin F2 alpha synthesis in bovine endometrium: role of calcium. AB - The objective of these experiments was to determine the role of Ca2+ during oxytocin-stimulated prostaglandin (PG) F2 alpha release from bovine endometrial tissue in vitro. Uteri were collected from dairy cows on the day after spontaneous luteal regression. Caruncular endometrial explants were dissected and incubated in vitro to determine phospholipase C activity or PGF2 alpha release. A23,187 (a calcium ionophore) and maitotoxin (an activator of voltage-gated L type calcium channels) stimulated release of PGF 2 alpha in a concentration dependent manner (P < 0.05). Thapsigargin (induces accumulation of Ca2+ in the cytoplasm by inhibiting endoplasmic reticulum Ca2+/ATPase pumps) stimulated release of PGF2 alpha in a concentration-dependent manner as well (P < 0.13). Oxytocin (10(-6) M), AIF4- (a nonspecific activator of G-proteins; 10(-5) M), A23,187 (10(-5) M), and melittin (a stimulator of phospholipase A2; 10(-4) M) stimulated PGF2 alpha release when explants were incubated in Ca(2+)-free medium (P < 0.10); however, oxytocin, A23,187, or melittin were unable to stimulate PGF2 alpha release when explants were incubated in Ca(2+)-free medium containing the calcium chelator EGTA (P < 0.10). This treatment did not prevent oxytocin or AIF4 from stimulating phospholipase C activity (P < 0.08). CoCl2 (a nonspecific Ca2+ channel blocker) and methoxyverapamil (a specific voltage-gated L-type Ca2+ channel blocker) prevented oxytocin from stimulating PGF2 alpha release (P < 0.05). Our results suggest that both extracellular and intracellular Ca2+ may be required for oxytocin to stimulate PGF2 alpha secretion in bovine endometrial tissue. PMID- 9861540 TI - Effects of steroid hormone treatment on mammary development in prepubertal heifers. AB - The objective of this study was to determine the effects of steroid hormone implantation in heifer calves on the ability of mammary tissue to develop subsequently in organ culture. Twenty-four calves were paired by date of birth and assigned to groups (eight calves/group). At 4, 7, or 10 mo of age, calves were implanted subcutaneously (s.c.) with pellets containing cholesterol or cholesterol, 17 beta-estradiol, and progesterone for 9 or 18 d. The calves were euthanized and uteri and mammary glands were removed and weighed. Slices of mammary parenchymal tissue were incubated for 5 d at 37 degrees C in a 50% O2, 5% CO2 humidified atmosphere in Waymouth's 752/liter medium supplemented with insulin (5.0 micrograms/ml) or lactogenic hormone complex insulin (5.0 micrograms/ml), aldosterone (0.1 microgram/ml), hydrocortisone (0.1 microgram/ml), and prolactin (1.0 microgram/ml) in the presence or absence of epidermal growth factor (EGF) (0.06 microgram/ml) to promote lobulo-alveolar development. Tissue sections were stained and mounted on slides for morphologic and histologic analysis or prepared to evaluate expression of beta-casein mRNA. There were no morphologic differences in slices from calf mammary tissues despite age, steroid hormone priming, or hormones used in tissue culture. The 4-mo-old calves required steroid priming followed by incubation of the tissue slices with the lactogenic complex with or without epidermal growth factor to induce cytological changes associated with lactogenesis but did not express beta-casein mRNA. At 7 mo of age, steroid hormone priming was not necessary for induction of alveolar formation and secretion. Incubation of the tissue slices from 7-mo-old calves with the lactogenic complex was sufficient to induce alveolar formation and secretion. However, beta-casein mRNA was not expressed. At 10 mo of age, exposure of tissue from calves to the lactogenic hormones caused histologic changes reminiscent of the ability to secrete milk regardless of hormone priming. However, estrogen and progesterone priming was necessary before incubation of the tissue slices with the lactogenic hormones to induce beta-casein mRNA expression. When epidermal growth factor was added to the lactogenic hormone complex, beta casein mRNA expression decreased. These data support the concept that there is a sequential development of responsiveness of the mammary gland to various hormones. By 10 mo of age, prepubertal heifers reach a stage of maturity where steroid hormone priming followed by incubation of tissue slices with the lactogenic hormones is sufficient to induce both structural and functional differentiation. PMID- 9861541 TI - Heat influences on plasma insulin and glucagon in response to secretogogues in non-lactating dairy cows. AB - Four non-lactating cows were offered a maintenance diet of hay wafer and a commercial concentrate. They were housed in a thermoneutral (TN; 20 degrees C) and then a hot (30 degrees C) environment in an artificial climate chamber. Glucose, arginine, butyrate, and insulin were administered through one jugular catheter, and from a catheter on the other side venous blood was collected. The peak increments in plasma insulin after the glucose and butyrate administrations were lower during heat exposure. The response of insulin after arginine injection was smaller in the hot compared with the thermoneutral environment; however, arginine injection resulted in a significantly higher secretion of glucagon in the hot environment. The response area of insulin after the insulin injection was smaller in the hot environment; however, insulin clearance rate was not changed. It is concluded that in non-lactating cows, insulin release is probably lower during heat exposure. With respect to plasma glucose during heat exposure, the lower basal values, lower concentrations after the end of the glucose infusion, and delayed recovery to basal values after the butyrate and insulin administrations observed, may indicate lower gluconeogenesis and glycogenolysis in the hot environment. PMID- 9861542 TI - Temporal effects of estradiol (E) on luteinizing hormone-releasing hormone (LHRH) and LH release in castrated male sheep. AB - We tested the hypothesis that rapidly expressed inhibitory effects of estradiol (E) on luteinizing hormone (LH) release in the male are attributable, in part, to suppression of luteinizing hormone-releasing hormone (LHRH) release. Hypophyseal portal cannulated, castrated male sheep were infused with E (15 ng/kg/hr) or vehicle. Portal and jugular blood samples were collected at 10-min intervals for 4 hr before, and for either 12 hr (E, n = 4; vehicle, n = 4) or 24 hr (E, n = 8; vehicle, n = 3) after the start of infusion. In animals sampled for 16 hr, temporal changes in both LHRH and LH were assessed. In animals sampled for 28 hr, only LH data were analyzed. Before either the 12-hr or 24-hr infusion, LHRH and/or LH mean concentrations, pulse amplitude and interpulse interval (IPI) did not differ between E- and vehicle-infused animals. In animals sampled for 16 hr, no effects of time or steroid x time interactions were detected for mean LHRH and LHRH pulse amplitude; however, both were greater (P < 0.01) in vehicle-infused than in E-infused males. LHRH IPI was unaffected by infusion. In contrast, both mean LH and LH pulse amplitude declined (P < 0.01) within 4-8 hr after the start of E infusion, whereas mean LH IPI was unaffected. In animals sampled for 28 hr, an effect of time (P < 0.01) and a steroid x time interaction (P < 0.01) was detected for mean LH, and there was an effect of time (P < 0.01) on LH pulse amplitude. Mean LH IPI was not affected. Our results show that in male sheep E rapidly reduces LH release in the absence of a detectable change in LHRH release. PMID- 9861543 TI - Development and validation of an improved enzyme-linked immunosorbent assay for the detection of thyroglobulin autoantibodies in canine serum samples. AB - An enzyme-linked immunosorbent assay to detect thyroglobulin autoantibodies (TGAB) in canine serum was developed and validated. The test result for each sample was derived from the optical density readings (OD) and expressed as an Ab score(%) calculated from three in-house calibrators. The assay specifically detected TGAB as judged from lack of response in the assay after samples had been incubated with specific antigen. Intra- and interassay coefficients of variation ranged from 2.0-4.9% and 4.6-9.9%, respectively. The detection limit, an Ab-score of 5.6%, was close to the median Ab-score of 10% observed in healthy dogs (n = 132). The median Ab-score of dogs with primary hypothyroidism and lymphocytic thyroiditis (n = 11), skin diseases (n = 35), and non-thyroidal diseases (n = 63) was 340%, 12%, and 8%, respectively. The prevalence of TGAB in hypothyroid dogs with lymphocytic thyroiditis (sensitivity) was 91% (95% confidence limits: 59% 99%). In dogs with dermatological diseases without lymphocytic thyroiditis the prevalence of TGAB was 3% corresponding to a specificity of 97% (95% confidence limit: 85%-100%). In dogs with non-thyroidal diseases and healthy dogs the prevalence of TGAB was 5% and 6%, respectively. The diagnostic accuracy of serum TGAB was evaluated by subjecting the data from 11 dogs with lymphocytic thyroiditis and 35 control dogs without lymphocytic thyroiditis to receiver operating characteristic curve analysis. The area under the receiver-operating characteristic curve (W = 0.966; 95% confidence limit 87%-100%) was significantly higher than that of a worthless test (0.5) (P < 0.0001), thereby indicating that serum TGAB measurements distinguished between dogs with and without lymphocytic thyroiditis. PMID- 9861544 TI - The thyrotropin (TSH) receptor: interaction with TSH and autoantibodies. PMID- 9861545 TI - Pathophysiology of the neuroregulation of growth hormone secretion in experimental animals and the human. AB - During the last decade, the GH axis has become the compelling focus of remarkably active and broad-ranging basic and clinical research. Molecular and genetic models, the discovery of human GHRH and its receptor, the cloning of the GHRP receptor, and the clinical availability of recombinant GH and IGF-I have allowed surprisingly rapid advances in our knowledge of the neuroregulation of the GH-IGF I axis in many pathophysiological contexts. The complexity of the GHRH/somatostatin-GH-IGF-I axis thus commends itself to more formalized modeling (154, 155), since the multivalent feedback-control activities are difficult to assimilate fully on an intuitive scale. Understanding the dynamic neuroendocrine mechanisms that direct the pulsatile secretion of this fundamental growth promoting and metabolic hormone remains a critical goal, the realization of which is challenged by the exponentially accumulating matrix of experimental and clinical data in this arena. To the above end, we review here the pathophysiology of the GHRH somatostatin-GH-IGF-I feedback axis consisting of corresponding key neurotransmitters, neuromodulators, and metabolic effectors, and their cloned receptors and signaling pathways. We propose that this system is best viewed as a multivalent feedback network that is exquisitely sensitive to an array of neuroregulators and environmental stressors and genetic restraints. Feedback and feedforward mechanisms acting within the intact somatotropic axis mediate homeostatic control throughout the human lifetime and are disrupted in disease. Novel effectors of the GH axis, such as GHRPs, also offer promise as investigative probes and possible therapeutic agents. Further understanding of the mechanisms of GH neuroregulation will likely allow development of progressively more specific molecular and clinical tools for the diagnosis and treatment of various conditions in which GH secretion is regulated abnormally. Thus, we predict that unexpected and enriching insights in the domain of the neuroendocrine pathophysiology of the GH axis are likely be achieved in the succeeding decades of basic and clinical research. PMID- 9861546 TI - The cytogenesis and pathogenesis of pituitary adenomas. PMID- 9861547 TI - Adrenocorticotropin insensitivity syndromes. PMID- 9861548 TI - Ethanol consumption in rats when dose size is under subject control. AB - Rats orally self-administered ethanol when they could control dose size as follows: A lever press initiated a trial; a press on 1 lever increased the previous trial duration by 30% whereas a press on a 2nd lever decreased the previous trial duration by 30%. During a trial, rats could drink either water, 8%, or 16% (wt/vol) ethanol (ETOH) from a lick-sensitive fountain. Mean per session intake of 8% and 16% ETOH was 0.8 g/kg and 1.3 g/kg, respectively. Water intake was negligible. Rats did not show a tendency to maintain a particular trial duration, nor did they adjust intertrial intervals to previous dose size (number of licks). When trials were collected into bouts, a positive relationship was found between bout size and interbout interval; this relationship was statistically significant for the 8% but not the 16% ETOH condition. PMID- 9861549 TI - Naloxone blocks reinforcement but not motivation in an operant runway model of heroin-seeking behavior. AB - The present study examined the effects of opiate receptor antagonism on both the motivation to seek heroin and the reinforcing consequences of heroin administration. Subjects were trained to discriminate between olfactory cues predicting either the delivery of intravenous heroin reinforcement (S+) or saline (S-). Subjects were then tested in the presence of the opiate receptor antagonist, naloxone (0.5, 1.0, or 3.0 mg/kg intraperitoneally). Naloxone had no effect on either S+ or S- trials. However, 24 hr later on the first posttreatment trial, subjects that had received heroin in the presence of naloxone (on the previous trial) now traversed the alley more slowly when presented with the S+. These data suggest that although the motivation to seek heroin was not disrupted by naloxone, the reinforcing consequences of heroin administration were. PMID- 9861550 TI - Chlordiazepoxide attenuates activity-induced anorexia and weight loss in rats. AB - In Experiment 1, the effect of repeated injections of 2.5, 5.0, or 10.0 mg/kg of chlordiazepoxide (CDP) on food intake and body weight was studied in rats on an activity anorexia (AA) regimen. For several days before CDP testing began, rats lived in activity wheels and had one 60-min meal per day. During CDP testing, this regimen continued except that each rat was injected with an appropriate dose of CDP or saline 30 min before each meal. CDP enhanced food intake; 5.0 mg/kg seemed most effective. However, the CDP-induced increase in eating did not noticeably stem weight loss. In Experiment 2, after several days of AA training, CDP (5.0 mg/kg) was tested under less severe conditions; food remained restricted, but access to the wheels was discontinued. Rats given CDP ate more and gained more weight than controls. These findings suggest that benzodiazepines such as CDP may help in treating anorexia nervosa and other anorectic conditions in humans. PMID- 9861551 TI - Comparison of acute behavioral effects of sustained-release and immediate-release methylphenidate. AB - The rate of onset of a drug's effect is an important determinant of its abuse potential. This experiment examined the acute behavioral effects of orally administered sustained-release methylphenidate (SR; 20-40 mg), immediate-release methylphenidate (IR; 20-40 mg), and placebo in 10 healthy volunteers. Drug effects were assessed before drug administration and periodically afterwards for 6 hr using drug-effect questionnaires and performance measures that are sensitive to the acute effects of stimulants. The IR formulation produced stimulant-like drug effects (e.g., increased ratings of "good effects") that generally varied as a function of dose and time. The SR formulation produced only transient effects on these measures. These findings are consistent with previous research on the influence of rate of onset using other drugs and suggest that the abuse potential of IR methylphenidate may be greater than that of SR methylphenidate. PMID- 9861552 TI - Discriminative and participant-rated effects of methylphenidate in children diagnosed with attention deficit hyperactivity disorder (ADHD). AB - Despite the demonstrated beneficial effects of methylphenidate and d-amphetamine for the treatment of attention-deficit hyperactivity disorder (ADHD), the discriminative and subjective effects of these compounds in children are not well understood. This study was designed to characterize such effects in children diagnosed with ADHD. In a series of 3 experiments, 17 children were examined to determine whether methylphenidate (n = 12) and d-amphetamine (n = 5) could be reliably discriminated at doses typically used in clinical practice. Under some conditions (e.g., when they were instructed to attend to the drug effects or when a wide range of doses was used), children discriminated methylphenidate (5.0-30.0 mg) from placebo. Children tested under a range of doses of d-amphetamine (2.5 20.0 mg) were unable to discriminate this drug from placebo reliably. Neither methylphenidate nor d-amphetamine produced reliable participant-rated effects. PMID- 9861553 TI - Context modulates effects of nicotine abstinence on human cooperative responding. AB - The effects of ad libitum smoking, abstinence, and 0-, 2-, and 4-mg nicotine gum on human cooperative responding were examined. Participants were provided the opportunity to respond cooperatively or independently to episodes initiated by a computer-simulated other person. Participants could also initiate episodes that ostensibly provided the other person the opportunity to respond cooperatively or independently of the participant. Working cooperatively added points to both the participant's and other person's counters. Working independently added points only to the participant's counter. Results demonstrated that abstinence decreased cooperative responses during episodes initiated by the computer-stimulated other person. Relative to abstinence and placebo gum conditions, ad libitum smoking and administration of 2- and 4-mg nicotine gum increased these cooperative responses. No gender differences were observed. The number of cooperative episodes initiated by the participants was not affected significantly by the smoking or gum conditions. Nicotine increased reports of vigor and decreased abstinence engendered reports of depression, anger, confusion, and tension. The difference in the effects of nicotine abstinence on the 2 classes of cooperative responding demonstrates that the social contingency mediates the behavioral effects of abstinence. PMID- 9861554 TI - Effects of priming positive and negative outcomes on drinking responses. AB - The purpose of this investigation was to study the effects of priming positive and negative expectancy outcomes on the drinking responses of college students. Men and women (N = 64) were randomly assigned to 1 of 3 priming conditions: a positive expectancy outcome condition, a negative expectancy outcome condition, and a neutral (control) condition. Participants were exposed to a series of semantic primes corresponding to their condition and then asked to complete a beer taste-rating task. Planned comparisons revealed that the average ratio of beer consumed to body weight in the positive condition was significantly greater than the average ratio in the neutral condition, and the average ratio of beer consumed to body weight was significantly less in the negative condition than the average ratio in the neutral condition. These findings are discussed as they relate to cognitive models of alcohol use. PMID- 9861555 TI - Effects of smokeless tobacco-related sensory and behavioral cues on urge, affect, and stress. AB - The hypothesis that smokeless tobacco-related sensory and behavioral cues can act as conditioned stimuli was tested in a counterbalanced double-blind experimental design. The nicotine content of snuff smokeless tobacco (ST) was manipulated for 24 male ST users by mixing ST with an ST substitute. Affect was manipulated through imagery scripts, stress was induced by a mental arithmetic task, and physiological measures and self-reported affect, stress, and urge for ST were collected. Urge for ST was consistently reduced regardless of the nicotine content in the ST conditions. Urge was increased by the stress manipulation and by negative affect when compared with positive affect. Urge for ST was positively correlated with stress and negative affect but was not correlated with positive affect. Physiological arousal was not related to urge. Results generally parallel studies of smoking and suggest that ST substitute products may aid ST cessation. PMID- 9861556 TI - Adults seeking treatment for marijuana dependence: a comparison with cocaine dependent treatment seekers. AB - Sixty-two individuals seeking treatment for marijuana dependence completed a comprehensive assessment. Sociodemographics, substance use, psychosocial functioning, psychiatric symptoms, and medical status were compared with similar data collected from 70 treatment-seeking, cocaine-dependent individuals. Substantial psychosocial and psychiatric problems were observed in both groups. In general, the marijuana group reported substance-use histories and a range of impairment comparable with the cocaine group; however, they showed less severe dependence. The marijuana group was more ambivalent and less confident about stopping their marijuana use than the cocaine group was about stopping their cocaine use. These findings indicate that treatment-seeking, marijuana-dependent individuals exhibit substantial problems and that further efforts to develop effective treatments for this population are warranted. PMID- 9861557 TI - The influence of an instruction on the stimulus effects of drugs in humans. AB - Participants discriminated between tripelennamine and placebo in experiments differing in instructional set. In 1 experiment (SED), participants were told that 1 of the 2 drugs was more sedative-like, and during the other (STIM), 1 was more stimulant-like. During generalization tests, participants received diazepam or d-amphetamine. Percent correct was the same in both experiments. Tripelennamine increased sedative and decreased stimulant effects. Amphetamine and diazepam produced typical subjective effects. Some subjective effects differed across experiments with more sedative and less stimulant effects during SED than STIM. In SED and STIM, capsules were labeled 80% of the time as a sedative and stimulant, respectively. Thus, instructions designed to give expectations had no effect on discrimination and only a few changes in subjective effects. When asked to name the drug that they believed they received, labels reflected instructional set. PMID- 9861558 TI - Evolution of the clinical manifestations of infection during the course of febrile neutropenia in patients with malignancy. AB - The impact of a standardized set of diagnostic interventions on the further management of 968 episodes of fever in neutropenic cancer patients who did not respond to initial therapy was assessed prospectively. At the onset of fever, 65% of patients had no additional signs of infection, whereas skin and soft tissue infections were present in 12%, and clinical sepsis and gastrointestinal infections in 8% each. After 72 h, 41% of the fevers still remained unexplained. New foci of infection emerged in 11% of the cases involving mainly the lungs, skin and soft tissues, and urinary tract. The presence of a lower respiratory tract infection or a microbiologically defined infection of any sort was associated with higher mortality than other types of infection were. Changes in initial antibiotic therapy were based on the results of the diagnostic measures specified in the protocol in only 15% of the cases. PMID- 9861559 TI - Hepatitis TT virus infection in high-risk groups. AB - The novel hepatitis TT virus first described by a Japanese group has been reported to be parenterally transmitted and furthermore, to have been detected in patients with hepatitis of unknown etiology. Hence, in the present study its prevalence was investigated within groups at high risk for contracting blood borne viruses, such as individuals with chronic liver disease, intravenous drug users and recipients of blood and blood products, as compared to voluntary blood donors and pregnant women. To that end, DNA was extracted from sera obtained from the respective patients and subjected to PCR using semi-nested primers. The frequency of TTV DNA detected within high risk groups, such as nine out of 50 patients with chronic non-A-to-G liver disease (18%), nine out of 98 hepatocellular carcinoma cases (9.2%), 17 out of 52 intravenous drug users (32.7%), 15 out of 80 thalassemia patients with multiple blood transfusions (18.8%) and three out of 31 prostitutes (9.7%) exceeded that among voluntary blood donors and pregnant women, which amounted to 14 out of 200 (7%) and seven out of 103 (6.8%), respectively. Additional molecular research should be performed in order to determine its short-, as well as long-term clinical significance. PMID- 9861560 TI - A prognostic score for postherpetic neuralgia in ambulatory patients. AB - The main objective was to develop a scoring system for easy use by the physician in daily clinical practice in deciding the appropriate treatment for his herpes zoster patient. Data from 635 patients who did not receive antiviral therapy were included in this analysis. Of these, 131 developed postherpetic neuralgia (PHN). Of the 29 variables tested univariately in this study, 15 showed a significant correlation with the incidence of PHN, but only six proved to contribute to the overall predictive power in the multivariate approach. Using two independent approaches, the model showed a very satisfactory performance in the validation sample. Patients without acute pain rarely developed PHN. In those with acute pain, being female, being over 50 years of age, having more than 50 lesions, having lesions of a hemorrhagic nature, having cranial or sacral localisation of the rash or having pain in the prodromal phase proved to be significant, multivariate factors. An easy-to-use scoring system used in a risk graph is proposed. These data should be useful in the individual treatment decision as well as in the design and analysis of therapeutic trials in herpes zoster. PMID- 9861561 TI - A proposal for the reliable culture of Borrelia burgdorferi from patients with chronic Lyme disease, even from those previously aggressively treated. AB - Since culture of Borrelia burgdorferi from patients with chronic Lyme disease has been an extraordinarily rare event, clarification of the nature of the illness and proving its etiology as infectious have been difficult. A method for reliably and reproducibly culturing B. burgdorferi from the blood of patients with chronic Lyme disease was therefore sought by making a controlled blood culture trial studying 47 patients with chronic Lyme disease. All had relapsed after long-term oral and intravenous antibiotics. 23 patients with other chronic illness formed the control group. Positive cultures were confirmed by fluorescent antibody immuno-electron microscopy using monoclonal antibody directed against Osp A, and Osp A PCR. 43/47 patients (91%) cultured positive. 23/23 controls (100%) cultured negative. Although persistent infection has been, to date, strongly suggested in chronic Lyme disease by positive PCR and antigen capture, there are major problems with these tests. This new method for culturing B. burgdorferi from patients with chronic Lyme disease certainly defines the nature of the illness and establishes that it is of chronic infectious etiology. This discovery should help to reestablish the gold standard in laboratory diagnosis of Lyme disease. PMID- 9861562 TI - Intralymphatic interleukin-2 in combination with zidovudine for the therapy of patients with AIDS. AB - In a pilot study the safety and therapeutic effects of an immunostimulatory intralymphatic treatment with natural human interleukin-2 (IL-2) in combination with zidovudine were evaluated in nine patients with AIDS. Therapy with IL-2 consisted of one subcutaneous injection of 0.1 microgram/kg IL-2, followed by four intralymphatic IL-2 infusions of 0.1 microgram/kg each within a period of up to 15 days. Enlargement of lymph nodes was seen in six and a transient increase of CD4 cells in five out of nine persons in association with the IL-2 therapy. An increase of HIV p24-antigenemia was observed only in the two patients in whom zidovudine dosage had to be reduced because of side effects. Moderate clinical side effects occurred in eight of the nine patients. Four patients developed zidovudine associated anemia. Six participants showed a favourable course of disease with survival of 25 to 54 months (median 30 months) despite a previous diagnosis of manifest AIDS before IL-2 therapy. This pilot study demonstrates that a combination therapy with intralymphatic IL-2 and zidovudine can induce positive immunomodulatory effects, even in the presence of manifest AIDS. Further studies should explore the tolerability and effects of a prolonged therapy with IL-2 in combination with a more potent antiviral drug combination therapy. PMID- 9861563 TI - Is there an effect of immunoglobulins and G-CSF on neutrophil phagocytic activity in preterm infants? AB - The percentage of neutrophils phagocytosing group B streptococci (GBS) in vitro was determined in ten healthy preterm infants (< 32 weeks of gestation) and adult controls by using an acridine orange fluorescence whole blood assay. When GBS were opsonized with adult serum, no difference in phagocytic activity was found between both groups after 10 and 30 min (preterms: 40% and 68%, adults: 32% and 56%, respectively). Phagocytosis rates in preterm infants decreased significantly to 6% and 18% (at 10 and 30 min) when pool serum of preterm infants was used instead. Supplementation of the preterm serum with either intravenous immunoglobulin or IgM-enriched immunoglobulin did not change the results significantly. The addition of granulocyte colony-stimulating factor (G-CSF) accelerated phagocytosis significantly after 10 min, but did not increase the overall phagocytic activity after 30 min in either group. Hence the potential benefits of intravenous immunoglobulins and G-CSF in neonatal sepsis may not be attributable to an immediate increase in and direct effect on neutrophil phagocytic activity. PMID- 9861564 TI - Colonization and infection with fluoroquinolone-resistant Escherichia coli among cancer patients: clonal analysis. AB - Escherichia coli with high-level fluoroquinolone resistance were isolated from feces and/or various body sites of 16 cancer patients who were on oral fluoroquinolone prophylaxis. Population analysis of fecal isolates in 11 patients showed that fluoroquinolone-resistant E. coli was the only aerobic gram-negative bacillus present and exhibited a relatively homogenous fluoroquinolone MIC distribution. Molecular typing by pulsed field gel electrophoresis of chromosomal DNA digests or by random amplified polymorphic DNA fingerprinting confirmed the clonal nature of gastrointestinal tract colonization with E. coli. Genotyping of ten colonies picked from the same fecal culture demonstrated identical strains in four of four patients examined. Identical genotypes from the same patient were isolated over prolonged periods of time in 12 of 12 cases examined, with one patient (with the longest follow-up of 14 months) who lost his initial genotype and became persistently colonized with a new genotype. In the 11 patients who developed infection due to fluoroquinolone-resistant E. coli, molecular typing also indicated that fecal colonization was associated with, and presumably preceded infection due to an indistinguishable genotype of fluoroquinolone resistant E. coli. PMID- 9861565 TI - Adverse reactions to tick-borne encephalitis vaccine: FSME-Immun. AB - Vaccination against tick-borne encephalitis with FSME-Immun vaccine was started in the Department of Infectious Diseases, University Medical School of Bialystok, Poland, in 1992. No serious adverse reactions after vaccine administration were observed. Post-vaccine side effects were reported in 242 (11.3%) persons after the first dose (n = 2,135) and only in 14 patients (1.2%) after the second one (n = 1,183). These effects were mild and transitory. No relationship was observed between the frequency of adverse reactions, general or local, and the initial anti-TBE virus antibody titres or the age of the immunized individuals. Post vaccine side effects were reported significantly more frequently among people not bitten by ticks. PMID- 9861566 TI - A survey of the use of teicoplanin in patients with haematological malignancies and solid tumours. AB - A significant number of open and comparative studies have now addressed the use of teicoplanin in the treatment of documented or presumed infection in patients with haematological and non-haematological malignancy. Available evidence suggests that teicoplanin is an effective agent against such infections, with an excellent safety profile. The use of teicoplanin and vancomycin may be justified as part of the initial management of clinically infected right atrial catheters in patients with malignancy. The first-line use of glycopeptides may also be appropriate in units where streptococcal and methicillin resistant staphylococcal infections are prevalent. However, such a policy should be reviewed regularly. Except in the above situations, a delay in the introduction of either teicoplanin or vancomycin in cancer patients does not appear to produce any excess mortality, but there may be some additional morbidity in terms of fever and malaise. The introduction of glycopeptides as second-line agents is indicated for sensitive, microbiologically documented infections and for patients who have not responded to empirical, first-line therapy. Non-inpatient treatment with teicoplanin is an area of ongoing interest and may be justified on both humanitarian and pharmacoeconomic grounds. The use of glycopeptides in the prophylactic setting remains controversial and should be avoided while the emergence of resistance, particularly in enterococci, should be monitored closely. PMID- 9861567 TI - Pharmacokinetic analysis of amikacin twice and single daily dosage in immunocompromised pediatric patients. AB - Ten children received amikacin twice daily and 13 were treated using the single daily protocol. All had fever and neutropenia on admission, and received a total daily dose of 20 mg/kg when included in the study. Individual pharmacokinetic parameters were calculated using a one-compartment model for two blood amikacin samples. The mean (+/- SD) of elimination half-life (h), amikacin clearance (l/h/kg), volume of distribution (l/kg), peak concentration (microgram/ml) and trough concentration (microgram/ml) were: 2.51 (0.74) and 2.85 (0.32) h; 0.26 (0.16) and 0.115 (0.02) l/h/kg; 0.74 (0.44) and 0.47 (0.11) l/kg; 19.1 (12.3) and 42.6 (12.6) micrograms/ml; 0.85 (0.74) and 0.18 (0.24) microgram/ml with twice and single daily dosage schedules, respectively. A single daily dose of amikacin had a significantly longer elimination half-life, lower clearance, higher peak concentration and lower trough concentration in comparison to the twice-daily schedule. The use of amikacin 20 mg/kg daily delivered in a single daily dose is recommended for immunocompromised pediatric patients with fever and neutropenia, in spite of the measured pharmacokinetic differences. PMID- 9861568 TI - Necrotizing fasciitis caused by Vibrio vulnificus: first published infection acquired in Turkey is the second time a strain is isolated in Germany. AB - Vibrio vulnificus, a marine vibrio, has recently been recognized as a potential human pathogen. It causes human infections with mortality rates up to 60%. Until 1991, most human isolations were reported from the USA, Japan and Taiwan. The second strain isolated in Germany is documented and a significant case of V. vulnificus infection acquired in Turkey is published for the first time. PMID- 9861569 TI - Meningoencephalitis caused by human herpesvirus-6 in an immunocompetent adult patient: case report and review of the literature. AB - Human herpesvirus-6 (HHV-6) is the etiologic agent of roseola infantum, and has been implicated as a possible cause of encephalitis in pediatric and adult patients. A case of meningoencephalitis in an otherwise healthy, immunocompetent 59-year-old woman is described. The diagnosis of HHV-6 meningoencephalitis was confirmed by detecting viral DNA in cerebrospinal fluid collected in the acute stage of the disease by polymerase chain reaction. The patient was treated with acyclovir and recovered without any sequelae. The current knowledge of the pathophysiology, clinical course and outcome of HHV-6 meningoencephalitis in immunocompetent adult patients is also reviewed. PMID- 9861570 TI - Haemophilus aphrophilus meningitis complicated by hydrocephalus in an immunocompetent adult. AB - A case of meningitis due to an unusual organism, Haemophilus aphrophilus in an adult is described. In spite of rapid clearance of the organism from the cerebrospinal fluid with intravenous cefotaxime, fever persisted over 8 days of therapy and infection was complicated by hydrocephalus. H. aphrophilus meningitis is uncommon and has not been previously reported in an adult male. PMID- 9861571 TI - Clearance of hepatitis G virus infection in thalassaemic children. PMID- 9861572 TI - Transmission of hepatitis E in Germany. PMID- 9861573 TI - Gaps in antidepressant prescribing in primary care in the United Kingdom. AB - An important determinant for achieving efficacy results in clinical practice comparable to those demonstrated in clinical trials is whether or not patients take their medication as prescribed. Recent studies have shown that 30-60% of patients do not take their medications as prescribed. Gaps between antidepressant prescriptions raise questions about the possibility of periods of nonadherence to medication in clinical practice. The purpose of conducting this study was to assess the likelihood of experiencing a gap of > 15 days between antidepressant prescriptions for patients with a depression-related diagnosis and to assess whether this likelihood varied across different antidepressants with tricyclic antidepressants and selective serotonin reuptake inhibitors. Episodes of antidepressant treatment were constructed using the Doctors' Independent Network general practitioner medical records database. For all antidepressant agents considered, approximately 50% of patients had a gap between prescriptions and 15 25% of patients had a gap of > 15 days between prescriptions. A significant proportion of patients in a general practitioner setting in the UK have gaps recorded of > 15 days between antidepressant prescriptions. Gaps between prescriptions raise the question of whether patients may be at risk for clinical consequences associated with nonadherence to therapy, such as reduced effectiveness or treatment interruption symptomatology. PMID- 9861574 TI - A comparison of descriptive characteristics of male outpatients and inpatients with affective disorders. AB - Recent studies of patients with affective disorders have found that there are biological differences between inpatients and outpatients. Concerned by these findings, we compared individuals admitted to our inpatient and outpatient affective disorders clinical research center who met criteria for major depression. We hypothesized that inpatients would be more severely ill, more suicidal, more functionally impaired, and have more co-morbid disorders and higher ratings of depression and mood state dysfunction. The demographic profiles, lifetime co-morbid Axis I diagnoses, consumption histories, symptom profiles, global assessment of functioning, and severity of current stressors (Axis IV) were compared and contrasted for the two groups. Inpatients had more severe current psychosocial stressors, lower current levels of functioning, increased lifetime co-morbid Axis I diagnoses, and increased rates of psychiatric hospitalizations, however, they did not have higher depression symptom ratings. In conclusion, inpatients and outpatients differed significantly in the severity of their stressors, coping abilities and history of previous hospitalizations, but not in most demographic variables or their current symptoms of depression. PMID- 9861575 TI - Risperidone and olanzapine: optimal dosing for efficacy and tolerability in patients with schizophrenia. AB - Schizophrenia is a chronic and debilitating disorder whose effective pharmacological management is often less than optimal. For several decades, pharmaceutical treatment for this disorder consisted of conventional neuroleptics such as haloperidol and chlorpromazine. However, the limitations of these drugs have driven the development of newer antipsychotics that are designed to be more efficacious and more tolerable than conventional agents. Newer agents available for consideration as first-line treatment options now include risperidone, olanzapine, sertindole and, more recently, quetiapine. Proper dosing has emerged as a vital factor in the effective use of these newer drugs. This report examines data derived from clinical trials and market research with risperidone and olanzapine to help clinicians determine the appropriate dose for efficacy and to appraise the adverse events associated with that efficacious dose. Current information suggests that, for most patients with schizophrenia, the optimal dose with respect to efficacy and tolerability of risperidone is < or = 6 mg/day. The optimal dose of olanzapine is less clear and may be 15 mg/day or higher. With the advent of these newer antipsychotics, clinicians now have more treatment options for the management of patients with psychotic disorders. Knowledge gained through clinical experience is needed to augment clinical trial results and to help define the most effective use of each of these agents. PMID- 9861576 TI - Cardiac side-effects of two selective serotonin reuptake inhibitors in middle aged and elderly depressed patients. AB - Selective serotonin reuptake inhibitors (SSRIs) are the 'new' drugs of first choice for the treatment of depression in the older patient. Systematic studies on the effects of SSRIs on cardiac function are scarce, despite the high prevalence of cardiac disorders in the older depressed patient. This is a study which systematically assessed cardiac function by echocardiography in middle-aged and elderly depressed patients treated with SSRI. In a double-blind randomized trial, 20 patients were assigned to receive fluvoxamine 100 mg/day [DOSAGE ERROR CORRECTED] or fluoxetine 20 mg/day [DOSAGE ERROR CORRECTED] for 6 weeks. Cardiac function was assessed by left ventricle ejection fraction, aortic flow integral and early or passive/late or active mitral inflow, and electrocardiography. Neither SSRI significantly affected cardiac function. Compared with patients without a history of myocardial infarction and/or hypertension, patients with such a history showed a significant improvement in left ventricular ejection fraction. Despite our small study sample, these data indicate that both fluoxetine and fluvoxamine do not affect cardiac function adversely. PMID- 9861577 TI - Elevated serum creatine kinase activity in adolescent psychiatric inpatients on admission. AB - Studies in adults have indicated a significant relationship between high serum creatine kinase levels on admission and acute psychosis. However, data on children are sparse. The files of 183 hospitalized children and adolescents (93 boys, 90 girls) with severe psychiatric disorders were reviewed for serum creatine kinase activity on admission, psychomotor agitation, Clinical Global Impression Score, need for intramuscular injection, number of neuroleptic medications and presence of neuroleptic malignant syndrome. Serum creatine kinase levels > 201 IU/ml were considered abnormal. Boys had significantly higher creatine kinase activity than girls. Division of the cohort by diagnosis yielded significantly higher levels in those with schizophrenia, affective disorders and mental retardation. Higher levels were also associated with higher Clinical Global Impression score on admission, use of injections and physical restraint, and nonresponse to neuroleptic medication. There were no cases of neuroleptic malignant syndrome. This first large-scale investigation of serum creatine kinase activity in young psychiatric inpatients shows a significant association between high creatine kinase activity and acute psychosis, similar to that in adults. Furthermore, high creatine kinase levels on admission are predictive of the severity of the psychosis, but are not associated with neuroleptic malignant syndrome. Because psychotic adolescents with high admission creatine kinase levels tend to be nonresponders, clinicians should consider the early use of atypical antipsychotics in this subgroup. PMID- 9861578 TI - Beneficial effect of risperidone on sleep disturbance and psychosis following traumatic brain injury. AB - Severe disturbances of sleep architecture and circadian rhythms are common in traumatic brain injured patients; however, complete absence of the rapid eye movement sleep stage is very rare. We describe a brain injured patient with cognitive disturbances who developed severe alterations of sleep architecture, accompanied by paranoid and jealousy delusions. Following several trials with conventional antipsychotics his psychotic state stabilized but he continued to complain of insomnia and daytime fatigue. When treated with risperidone 2 mg/day, both his sleep and the delusional thoughts improved markedly and his daytime alertness increased. Severe deterioration of his support system brought about discontinuation of treatment with re-emergence of all symptoms. PMID- 9861579 TI - Non-fatal mirtazapine overdose. AB - We report the case of intoxication of a 41-year-old female patient suffering from major depression with mirtazapine complicated by severe hypothermia. The patient was brought to an intensive care unit 10 hours after intake of 1200 mg mirtazapine and 20 mg lorazepam and laying outside at outdoor temperatures below 0 degree C. The mirtazapine plasma-level was 368 ng/ml still after 41 hours. Body temperature was reduced to 26 degrees C and rhabdomyolysis due to hypothermia was observed. Cardial and respiratory function was compromised and intubation was necessary. The patient recovered without persisting somatic impairment. PMID- 9861580 TI - Weak and feeble bones no more: the National Osteoporosis Foundation speaks out. PMID- 9861581 TI - "Press 1 for more options". PMID- 9861582 TI - News from the Society for the Advancement of Women's Health Research. Herbal medicines: are supplements safe? PMID- 9861583 TI - Observations from the CDC. The urgent need for new HIV/STD prevention options for women. PMID- 9861584 TI - Patient education. Vision and eye care. PMID- 9861585 TI - Toward optimal health: the experts respond to the aging eye. Interview by Jodi Godfrey Meisler. PMID- 9861586 TI - Why physician gender matters in shaping the physician-patient relationship. AB - Societal values regarding the nature and consequences of patient autonomy and medical paternalism underscore the current debates surrounding informed consent and shared decision making. The debate is significant in that it both reflects and determines normative expectations for physician and patient conduct as well as the nature and form of the therapeutic relationship. Analysis of the literature describing communication differences between physicians of different genders indicates that female physicians show a greater affinity for collaborative models of patient-physician relationship than do their male colleagues. Female physicians spend more time with their patients, are more likely to engage their patients in discussions of their social and psychologic context, and deal more often with feelings and emotions. Moreover, female physicians facilitate partnership and patient participation in the medical exchange more effectively than do male physicians. The authors propose that the quality of the interactive process is critical to the establishment of a therapeutic relationship and that this process is related to physician gender. They also suggest that physician gender matters in the shaping of the patient physician relationship through this interactive process. PMID- 9861587 TI - A case study and national database report of progressive systemic sclerosis and associated conditions. AB - We report the case of a 34-year-old white woman with a history of progressive systemic scleroderma (PSS) and diffuse alveolar hemorrhage (DAH) that may be either a rare complication of PSS or induced by D-penicillamine. The DAH progressed to hemoptysis and led to intubation for airway protection. The patient progressed to acute renal failure. Her chest x-ray revealed diffuse bilateral infiltrates. She developed pulmonary fibrosis with secondary pulmonary hypertension. She experienced a brief period of improvement of her respiratory status after steroid treatment. We also report a database of 21,442 decedents with PSS over a 15-year period from 1979 to 1994. Our report demonstrates that of over 21,000 decedents, only 0.2% had pulmonary hemorrhage or hemoptysis or both listed as a cause of death. The data also demonstrate that PSS was the underlying cause of death more frequently in younger people. Age-adjusted mortality rates were higher for blacks than for whites and for women than for men. PMID- 9861588 TI - Prevalence of heart disease in older women in a nursing home. AB - The prevalence of coronary artery disease (CAD) and the incidence of new coronary events are similar in older men and women. Independent risk factors for new coronary events in older women include age, prior CAD, cigarette smoking, hypertension, diabetes mellitus, high serum total cholesterol and triglycerides, and low serum high-density lipoprotein cholesterol. Older women have a higher prevalence of hypertension than older men. In older women with hypertension, echocardiographic left ventricular hypertrophy is a powerful independent predictor of new coronary events, atherothrombotic brain infarction, and congestive heart failure (CHF). Older women have a higher prevalence of rheumatic mitral stenosis and of mitral annular calcium than older men. Older women and men have a similar prevalence of valvular aortic stenosis, aortic regurgitation, mitral regurgitation, hypertrophic cardiomyopathy, and idiopathic dilated cardiomyopathy. The prevalence and incidence of CHF increase with age. The prevalence of normal left ventricular ejection fraction associated with CHF increases with age and is higher in older women than in older men. The prevalence of chronic atrial fibrillation increases with age and is similar in older men and women. Atrial fibrillation is an independent predictor of new coronary events and thromboembolic stroke in older women. Older women with unexplained syncope should have 24-hour ambulatory electrocardiograms to determine whether pauses > 3 seconds are present, requiring permanent pacemaker implantation. PMID- 9861589 TI - A model for integrating women's health issues into a problem-based curriculum. AB - The process of incorporating new material into an existing medical curriculum frequently produces lengthy debate, political maneuvering, and competition for curricular time. The faculty of the Women's Health Education Program at MCP Hahnemann School of Medicine, developed a stepwise process, or framework, for including women's health teaching for students in the problem-based curriculum. This process can be applied to the integration of any body of information. The key elements of the process are to define the full scope of what needs to be taught, develop teaching objectives, identify opportunities to introduce the information into the curriculum, develop strategies that capitalize on existing curricula, enlist the collaboration of a broad range of key faculty, develop evaluation tools, and assess whether students have achieved the expected competencies. PMID- 9861590 TI - Prevalence of multiple cardiovascular disease risk factors among women in the United States, 1992 and 1995: the Behavioral Risk Factor Surveillance System. AB - We sought to examine the prevalence of self-reported multiple cardiovascular disease (CVD) risk factors (hypertension, high blood cholesterol, diabetes, overweight, and current smoking) among women in 1992 and 1995 in the United States using data from the Behavioral Risk Factor Surveillance System. In 1992, 37.5%, 34.4%, and 28.1% of women had zero, one, and two or more of the five risk factors, respectively. In 1995, the respective estimates were 35.5%, 34.3%, and 30%. In both years, the prevalence of two or more risk factors increased with age, decreased with educational level, was higher among black women (lowest among Hispanic women and women of other ethnic groups), and higher among women reporting cost as a barrier to healthcare. The percentage of women with two or more risk factors was higher in 1995 than in 1992 for 35 of 48 states, being statistically significant for 7 states. The percentage of women with at least two risk factors was not significantly lower in 1995 than in 1992 for any state. A higher percentage of women reported having multiple CVD risk factors in 1995 compared with 1992. A multifactorial approach to primary prevention and risk factor reduction should be encouraged to help reduce the prevalence and burden of CVD among women. PMID- 9861591 TI - Early life stress, negative paternal relationships, and chemical intolerance in middle-aged women: support for a neural sensitization model. AB - This study (ntotal = 35) compared early life stress ratings, parental relationships, and health status, notably orthostatic blood pressures, of middle aged women with low-level chemical intolerance (CI group) and depression, depressives without CI (DEP group), and normals. Environmental chemical intolerance is a symptom of several controversial conditions in which women are overrepresented, that is, sick building syndrome, multiple chemical sensitivity, chronic fatigue syndrome, and fibromyalgia. Previous investigators have postulated that people with CI have variants of somatization disorder, depression, posttraumatic stress disorder (PTSD) initiated by childhood abuse or a toxic exposure event. One neurobehavioral model for CI, somatization disorder, recurrent depression, and PTSD is neural sensitization, that is, the progressive amplification of host responses (e.g., behavioral, neurochemical) to repeated intermittent stimuli (e.g., drugs, chemicals, endogenous mediators, stressors). Females are more vulnerable to sensitization than are males. Limbic and mesolimbic pathways mediate central nervous system sensitization. Although both CI and DEP groups had high levels of life stress and past abuse, the CI group had the most distant and weak paternal relationships and highest limbic somatic dysfunction subscale scores. Only the CI group showed sensitization of sitting blood pressures over sessions. Together with prior evidence, these data are consistent with a neural sensitization model for CI in certain women. The findings may have implications for poorer long-term medical as well as neuropsychiatric health outcomes of a subset of women with CI. Subsequent research should test this model in specific clinical diagnostic groups with CI. PMID- 9861592 TI - Predictors of menopausal hot flashes. AB - There are limited data on the factors associated with menopausal hot flashes, a common and potentially morbid condition. The objective of this study was to identify predictors of menopausal hot flashes. To meet this objective, 233 naturally perimenopausal or post-menopausal women (ages 45-65) attending a large urban hospital center primary care clinic, mammography unit, or women's health practice were enrolled. The women responded to a self-administered questionnaire assessing selected demographic factors, reproductive history, and behavioral factors. Sixty-seven percent of respondents experienced hot flashes, with 63% reporting frequent hot flashes (at least one hot flash per day) and 60% with hot flashes describing the hot flashes as severe. Women with hot flashes were significantly more likely to have mothers who experienced hot flashes (OR = 4.4, CI = 2.0-10.0) or to be smokers (OR = 2.0, CI = 1.2-3.5). There were no statistically significant associations between hot flashes and other selected demographic, reproductive, or behavior characteristics. These results reveal that menopausal hot flashes are associated with a maternal history of hot flashes as well as with cigarette smoking. These results may help physicians to counsel their patients about smoking cessation. PMID- 9861593 TI - Magnesium supplementation alleviates premenstrual symptoms of fluid retention. AB - We investigated the effect of a daily supplement of 200 mg of magnesium (as MgO) for two menstrual cycles on the severity of premenstrual symptoms in a randomized, double-blind, placebo-controlled, crossover study. A daily supplement of 200 mg of Mg (as MgO) or placebo was administered for two menstrual cycles to each volunteer, who kept a daily record of her symptoms, using a 4-point scale in a menstrual diary of 22 items. Symptoms were grouped into six categories: PMS-A (anxiety), PMS-C (craving), PMS-D (depression), PMS-H (hydration), PMS-O (other), and PMS-T (total overall symptoms). Urinary Mg output/24 hours was estimated from spot samples using the Mg/creatinine ratio. Analysis of variance for 38 women showed no effect of Mg supplementation compared with placebo in any category in the first month of supplementation. In the second month there was a greater reduction (p = 0.009) of symptoms of PMS-H (weight gain, swelling of extremities, breast tenderness, abdominal bloating) with Mg supplementation compared with placebo. Compliance to supplementation was confirmed by the greater mean estimated 24-hour urinary output of Mg (p = 0.013) during Mg supplementation (100.8 mg) compared with placebo (74.1 mg). A daily supplement of 200 mg of Mg (as MgO) reduced mild premenstrual symptoms of fluid retention in the second cycle of administration. PMID- 9861594 TI - Frequency and response to vaginal symptoms among white and African American women: results of a random digit dialing survey. AB - The prevalence of vaginal symptoms and response to those symptoms in a nonclinic setting has not been previously described. Two thousand women living throughout the United States identified by random digit dialing completed a computer assisted telephone interview about history of vaginal symptoms and use of healthcare services in response to these symptoms. The analysis was limited to 1698 white (WA) and 144 African American (AA) women (n = 1842). An episode of vaginal symptoms of any severity during 1995 was reported by 7.5% of WA women and 18.1% of AA women. Fifty-five percent of WA women and 83% of AA women with symptoms consulted a healthcare professional about their most recent episode. The racial difference in prevalence and consultation was not explained by marital status, education, employment, or lifetime number of sex partners. Most women purchased an over-the-counter antifungal preparation to treat their symptoms, whether or not a physician was consulted. The racial differences in prevalence and use of health services in response to vaginal symptoms observed here should be confirmed, and the potential causes should be explored. PMID- 9861595 TI - Women's health literatureWatch. PMID- 9861596 TI - Acute atraumatic quadriparesis in a college football player. AB - Transverse myelitis is a rare neurologic disorder. It is an interruption of spinal cord function not caused by macrotrauma. Symptoms develop rapidly and consist of ascending paralysis, diminished or absent sensation below the cervical or thoracic region, and often urinary retention. Etiologies include parainfectious events, multiple sclerosis, autoimmune disorders, vascular insufficiency, paraneoplastic myelopathy, postvaccinial events, idiopathic occurrence, and minimal trauma. Treatment generally consists of supportive measures. The use of steroids to hasten recovery remains controversial but is routine in most cases. The time period and degree of recovery is variable. We present a case of rapid onset of neurologic symptoms in a college football player right before a game. No other sports related cases have been reported in the sports medicine literature. Diagnostic, therapeutic, and historical aspects of this rare but important disorder are discussed. PMID- 9861597 TI - Effects of weight loss and exercise training on natural killer cell activity in obese women. AB - PURPOSE: The purposes of this study were two-fold: (1) to evaluate the effects of an 8-wk weight loss program on natural killer (NK) cell activity in obese women and 2) to determine whether an additional program of combined aerobic and resistance exercise training modified the effects of caloric restriction on immune function. METHODS: Twenty-two healthy obese women with a mean weight of 96.9 +/- 14 kg and age of 38 +/- 7 yr were randomly assigned to diet-alone (D) or diet-plus-exercise training (D + EX) conditions. Subjects consumed 950 kcal.d-1 using prepackaged portion-controlled foods. Subjects in the D + EX group participated 3 times.wk-1 in a supervised program of light-to moderate-intensity aerobic activity and resistance training. Data were analyzed using a repeated measures ANOVA. RESULTS: After 8 wk of treatment, body weight decreased significantly in both groups (10.8% in D vs 11.4% in D + EX), whereas absolute and relative VO2peak increased in only D + EX (12.3% in D vs 57.7% in D + EX). Both groups experienced significant decreases in peripheral blood leukocytes and lymphocytes, although cell numbers remained within clinically normal range at week 8. NK cell (CD56+) proportion was unchanged in both groups after weight loss. The proportion of peripheral mononuclear cells expressing the interleukin-2 receptor-alpha (IL-2R alpha) (CD25+) decreased significantly (25.2%) in D and was unchanged in D + EX, resulting in a significant difference between groups at week 8. NK cell cytotoxicity was suppressed in D and unchanged in D + EX after treatment. Changes in NK cell activity were significantly correlated with proportional changes in (CD25+) (r = 0.584, P = 0.022), but not CD56+. CONCLUSIONS: A combined program of light- to moderate-intensity aerobic and resistance exercise offsets the apparent decrement in NK cell activity associated with weight loss. PMID- 9861598 TI - Gender effect on beta-endorphin response to exercise. AB - PURPOSE: Twelve healthy men (26.4 yr) and women (26.8 yr) were compared at rest and after cycling for 25 min at 60 and 80% VO2max to determine whether gender and menstrual cycle influenced circulating beta-endorphin concentration (BE). METHODS: VO2max was determined on a cycle ergometer, and subjects completed the exercise in a randomized order. Women were tested in both the luteal (L) and follicular (F) phases of their menstrual cycle, which was confirmed by their blood estrogen levels. All tests were conducted in the morning after a 30-min rest (12-h postabsorptive). An indwelling venous catheter placed in a forearm vein enabled blood sampling at rest, 25 min of cycling, and 25 min of recovery. RESULTS: Resting BE was similar for men before both 60 and 80% intensities of exercise, 5.27 +/- 0.43 and 5.30 +/- 0.33 pmol.mL-1, respectively. BE was not significantly changed at 60% VO2max (6.54 +/- 0.33 pmol.mL-1) but significantly increased at 80% VO2max (11.90 +/- 1.98 pmol.mL-1). Women tended to have slightly lower BE during the L compared with F, but this did not reach significance (L = 4.40 +/- 0.22, F = 4.73 +/- 0.30 pmol.mL-1). Cycling at 60% VO2max did not significantly increase BE in the L (5.41 +/- 0.42 pmol.mL-1) nor the F (5.35 +/- 0.40 pmol.mL-1). Cycling at 80% VO2max increased BE to a similar extent in both the L and F phase, respectively (10.44 and 10.96). Although the BE concentrations tended to be slightly lower in women compared with men at 80% VO2max, this did not reach statistical significance. CONCLUSIONS: These data suggest that women cycling at 80% VO2max will have a similar BE response to men independent of their menstrual cycle. BE in women at rest and who exercise at lower exercise intensities may have slightly lower BE levels then men independent of the time of the women's menstrual cycle. PMID- 9861599 TI - Effects of high fat versus high carbohydrate diets on plasma lipids and lipoproteins in endurance athletes. AB - PURPOSE AND METHODS: Recent research suggesting the performance benefits of high fat diets for endurance athletes have been viewed with caution because of the potential negative health consequences, including increased coronary heart disease risk. This study examined the effects of a high fat (HF: 50% of total energy from fat, 37% carbohydrate) versus a high carbohydrate (HC: 15% of total energy from fat, 69% carbohydrate) diet on plasma lipids and lipoproteins in 32 endurance trained cyclists over a 3-month period. Plasma total, low density lipoprotein (LDL), high density lipoprotein (HDL), HDL2 and HDL3 cholesterol, triglycerides, apolipoprotein A1, and hematocrit (Hct) were measured at baseline and after weeks 4, 8, and 12. RESULTS: Changes in lipids and lipoproteins from baseline to week 12 did not differ between the two groups except for triglycerides, which increased significantly from 1.04 +/- 0.17 mmol.L-1 to 1.28 +/- 0.31 mmol.L-1 in HC (P = 0.012). The only significant changes that occurred within each group from baseline to week 12 was the significant increase in total cholesterol and triglycerides in HC. Body composition changes did not differ between the two groups from baseline to week 12 as measured by dual x-ray absorptiometry. CONCLUSIONS: During periods of endurance training when energy requirements are high, increasing the percentage of fat in the diet to approximately 50% of total energy did not result in adverse changes to the plasma lipoprotein profiles of this group of athletes. PMID- 9861600 TI - Exercise training induced alterations in prepubertal children's lipid-lipoprotein profile. AB - PURPOSE: This study examined the effect of exercise training on prepubertal children's (ET, N = 28) lipid-lipoprotein profile, relative to a maturity matched control group (CON, N = 20). METHODS: Training for ET involved stationary cycling for 30 min, 3 times.wk-1 for 12 wk, at 79.3 +/- 1.2% (mean +/- SD) peak heart rate (HR). Controls maintained their usual lifestyle pattern. Plasma concentrations of total triacylglycerol (TG), total cholesterol (TC), and high density lipoprotein (HDL)-cholesterol (HDL-C) were determined pre- and postintervention. Low-density lipoprotein (LDL)- cholesterol (LDL-C) was subsequently estimated from these concentrations, and the ratios TC/HDL-C and LDL C/HDL-C were also calculated. There were no pretest differences (P > 0.05) for any of these blood analytes between groups. The following, potentially, confounding variables were also measured: peak VO2, percent body fat (%BF), dietary composition, and habitual physical activity. These variables, with pretest HDL-C, were included as covariates in two-way split plot ANCOVA analyses. Dietary variables were not included as covariates as they were not related to any of the blood analytes. RESULTS: There were no differences over time or between groups for TG and TC (P > 0.05). LDL-C decreased in ET (-10.2%) but remained unchanged in CON (0.3%) over the intervention period (P < 0.05). HDL-C increased in ET (9.3%) but decreased in CON (-8.9%) (P < 0.01). A similar, but inverted, pattern of change (P < 0.01) was revealed for both ratios, TC/HDL-C (-11.6% vs 6.3%, ET and CON, respectively), and LDL-C/HDL-C (-17.2% vs 8.0%, ET and CON, respectively). The favorable alterations in the lipid-lipoprotein profile for ET were independent of alterations in peak VO2 (group x time interaction, P < 0.05), %BF (main effect time, P < 0.01), and habitual physical activity (group x time interaction, P < 0.01). CONCLUSIONS: In conclusion, the favorable alterations in the lipoprotein profile seen in this study would suggest that it is possible to influence the prepubertal lipoprotein profile independent of alterations in confounding variables such as body composition, cardiorespiratory fitness, and habitual physical activity. PMID- 9861601 TI - A comparison of plasma glutamine concentration in athletes from different sports. AB - PURPOSES: The purposes of the current investigation were to compare resting plasma glutamine concentration in athletes from different sports and to determine the relationship between resting plasma glutamine concentration and dietary protein intake. METHODS: Resting plasma glutamine concentration was measured in five groups of eight distance runners, competitive swimmers, cyclists, powerlifters, and nonathletes. Dietary protein intake of each subject was measured (g.d-1 and g.kg-1.d-1). RESULTS: Plasma glutamine concentration was significantly different between sports (P = 0.000, ANOVA) with mean plasma glutamine concentration of cyclists significantly higher than in all other groups, and mean plasma glutamine concentration of powerlifters and swimmers significantly lower than in cyclists and nonathletes (P < 0.05, post hoc analysis). There was no significant relationship between plasma glutamine concentration and total dietary protein intake when expressed as g.d-1 (r = 0.11, P > 0.05); however, plasma glutamine concentration and dietary protein relative to body mass (g.kg-1.d-1) were significantly inversely correlated (r = -0.37, P = 0.007). CONCLUSIONS: These data suggest that resting plasma glutamine concentration may vary between sports, possibly due to metabolic demands of the different sports; dietary factors may also affect plasma glutamine concentration. PMID- 9861602 TI - Gender comparisons of the mechanomyographic responses to maximal concentric and eccentric isokinetic muscle actions. AB - PURPOSE: The purpose of this study was to determine whether there is a gender difference in the velocity-related patterns of mechanomyographic (MMG) responses to maximal isokinetic concentric (CON) and eccentric (ECC) muscle actions. METHODS: Adult males (N = 15) and females (N = 16) performed maximal CON and ECC muscle actions of the leg extensors on a calibrated Cybex 6000 dynamometer at velocities of 30, 90, and 150 degrees.s-1. MMG was detected by a piezoelectric crystal contact sensor placed over the vastus lateralis muscle. RESULTS: The results indicated that there were decreases in CON peak torque (PT) across velocities, while ECC PT remained constant with increasing velocity for both genders. MMG amplitude increased significantly (P < 0.05) with velocity in both the males and females for CON and ECC muscle actions. There was a gender difference in the velocity-related patterns of MMG responses to maximal isokinetic CON muscle actions; however, there was no gender difference in the pattern of ECC MMG responses. CONCLUSIONS: The gender difference in CON MMG responses may be attributed to the greater percent decline in CON PT across velocity for the females than the males. In addition, the males displayed greater CON and ECC MMG amplitudes at all muscle action velocities than the females, possibly because of gender differences in muscle mass and/or thickness of the adipose tissue layer. PMID- 9861603 TI - Effect of lower extremity muscular fatigue on motor control performance. AB - PURPOSE: The purpose of this study was to determine whether lower extremity fatigue affects the ability of an individual to balance on an unstable platform. METHODS: Twenty healthy subjects (average age, 29 yr, range, 20 to 39 yr) were tested on an instrumented balance assessment system. Static tests were done on the limbs unilaterally and then bilaterally, and finally a dynamic test was performed in which the subject moved the platform in a circular manner to chase a moving object on a computer screen. After testing, subjects were fatigued using an isokinetic dynamometer, which imposes closed kinetic chain antagonistic exercise on the ankle, knee, and hip, similar to a stair stepper. No rest was allowed, and subjects were fatigued to less than 50% of their initial tested force. Subjects were then immediately retested on the unstable platform using the same testing protocol. All subjects completed the testing. RESULTS: Analysis of pre- and post-fatigue balance results demonstrated significant decreases in motor control performance on the three static tests following exercise to fatigue in all subjects (P < 0.001). CONCLUSIONS: Our findings support anecdotal evidence that fatigued individuals are at increased risk of injury because of loss of balance. Avoidance of fatigue and preconditioning may prevent injury. PMID- 9861604 TI - Electromyographic analysis of grand-plie in ballet and modern dancers. AB - PURPOSE: The purpose of this report is to describe lower extremity muscle activity in grand-plie, as determined by EMG analysis; to compare and contrast muscle function in grand-plie and demi-plie to support the hypothesis that grand plie is not simply a deeper demi-plie, but rather a fundamentally different movement in terms of muscle use; and to present further evidence in support of the hypothesis that ballet dancers use muscles differently than modern dancers in dance movement. METHODS: Surface electromyography was used to analyze lower extremity muscle activity during grand-plie in first position with lower extremities turned out in five ballet and seven modern female professional dancers. RESULTS: Electromyographic (EMG) activity of tibialis anterior included continuous activity from heel-off during the lowering phase, through midcycle, and ending at heel-on during the rising phase in all grand-plies; the majority of tibialis anterior EMG tracings in ballet dancers had additional activity at the end of the rising phase. All EMG tracings for vastus lateralis and medialis included a peak of activity during the lowering phase, a decrease (valley) at midcycle, followed by another peak during the rising phase; increased activity at the end of the rising phase was observed in most grand-plie in ballet, and not modern, dancers. Adductor EMG activity was also observed in all tracings with a peak during the lowering phase from heel-off to midcycle, a valley at midcycle, followed by a peak of activity in early rising phase; the midcycle valley was of lower, and the rising phase peak of higher, activity in ballet compared with modern dancers. Variation of EMG patterns was observed for lateral and medial gastrocnemius, gluteus maximus, and hamstrings. CONCLUSIONS: The data support the concept that lower extremity muscle activity in dance movement is comprised of three major types: (a) unique, characteristic activity required for the execution of the movement; (b) varied activity which is characteristic of dancers of different dance idioms; and (c) varied activity which may depend on factors such as balance, personal habit, and individual training background. Furthermore, EMG activity of vastus lateralis and medialis at the midcycle valley in grand-plie was significantly less in ballet dancers than in modern dancers despite similar degree of knee flexion, suggesting that ballet dancers may have lower patellofemoral joint reaction force at midcycle than modern dancers. PMID- 9861605 TI - Importance of wash riding in kayaking training and competition. AB - PURPOSE: The use of different wash-riding techniques is common during kayak training and competition. Changes in wash-riding positions could imply a different exercise intensity. The aim of this study, therefore, was to quantify the energy savings made when a kayaker is "wash riding." METHODS: Eight male international flat water kayakers, who performed a field test of 2000 m in each of the four wash-riding positions, head (H), right wave (RW), left wave (LW), and end position (V), were studied. The data investigated were: time, stroke rate, blood lactate (BL), heart rate (HR), and rate of perceived exertion (RPE). Under laboratory conditions kayakers performed the same intensity of exercise in a kayak ergometer, and HR, oxygen uptake (VO2), BL, mean power output (W), and RPE were measured. RESULTS: The results show significant differences (P < 0.05) among H, RW/LW, and V. The mean values for BL (P < 0.05) were 4.2, 2.0, 2.2, and 1.5 mmol.L-1, for H, RW, LW, and V, respectively. RPE also revealed differences, with values of 15, 12.6, 12.6, and 9.7 for H, RW, LW, and V, respectively. Mean power output gave values of 190.3 (H), 155.6 (RW and LW), and 129.5 (V) W. HR was different between H and V (172 and 151), while stroke rate was different among the parameters H, RW, and V (93.7, 88.8, and 87.6, respectively). The VO2 in the kayak ergometer test showed a difference between H and V (3.78 and 2.23 L.min-1). CONCLUSIONS: We conclude that "wash riding" involves a saving in energy cost of between 18% and 31.9%, depending on the position. This conclusion is of importance for the quantification and calibration of kayak training and competition. PMID- 9861606 TI - The gender difference in distance running performance has plateaued: an analysis of world rankings from 1980 to 1996. AB - PURPOSE: Annual world rankings from 1980-1996 were used i) to determine if the gender difference in distance running events has stabilized, and ii) to examine whether the relative decline in pace as race distance increases is different between men and women. World best (WB) and 100th best times in the 1500 m and the marathon (42 km) for each of the 17 years were the indices used to represent overall trends in men's vis-a-vis women's distance running. METHODS: These data were analyzed using regression analyses to develop and compare gender-specific equations. In the 1500 m, the gender difference in WB times (11.1 +/- 1.1%) was consistent from 1980 to 1996, and the slight rate of improvement in event depth (i.e., 100th rankings) was similar for men and women. RESULTS: In the marathon, the gender difference in WB times (11.2 +/- 0.9%) was essentially the same as for the 1500 m. In 1980, the marathon was a fairly new event for the women, having only just been sanctioned by the International Amateur Athletics Federation (IAAF). As a consequence, the depth of the field increased quickly from 1980 to 1984. Since the mid-1980s, the rate of improvement for women in 100th-ranked times has leveled off to equal that of men. The average declines in relative pace for men and women from the 1500 m to the 10 km to the marathon were found to be remarkably similar with no diminishing of the gender difference as race distance increased. CONCLUSIONS: Based on worldwide indices of competitive distance running, the gender difference in distance running performance has plateaued in recent years. Concomitantly, over the past decade, opportunities for women to train and compete have approached parity with those of men. It is likely that the current gender difference in performance will remain fairly constant because of biological differences between men and women that give men an advantage in distance running. PMID- 9861608 TI - Effects of strength training on intra-abdominal adipose tissue in obese prepubertal girls. AB - PURPOSE: The purpose of this study was to determine the effects of strength training on fat distribution and its relationship to glucose tolerance in obese prepubertal girls. METHODS: A strength training intervention study was designed in which the children exercised three times per week for 5 months. Twelve healthy, obese prepubertal girls (ages 7-10 yr, > 95th percentile weight for height) were enrolled in the study. Body composition was measured by dual-energy x-ray absorptiometry and computed tomography, and glucose tolerance was measured by a 3-h oral glucose tolerance test at baseline and after training. RESULTS: Significant increases in height, weight, fat-free mass, fat mass, and subcutaneous abdominal adipose tissue occurred after training (P < 0.05), whereas intra-abdominal adipose tissue (IAAT) remained stable (N = 11). Insulin area was highly correlated with IAAT before (r = 0.91) and after (r = 0.90) training (both P < 0.01, N = 9). CONCLUSION: In growing obese prepubertal girls undergoing a strength training program, IAAT remains unchanged, whereas subcutaneous abdominal fat and total body fat increased; insulin area is related to IAAT in these children. PMID- 9861607 TI - Chromium picolinate effects on body composition and muscular performance in wrestlers. AB - PURPOSE: The purpose of this study was to assess the effects of 14 wk of chromium picolinate supplementation during the final 16 wk of a preseason resistance and conditioning program on body composition and neuromuscular performance in NCAA Division I wrestlers. During this phase of training, wrestlers are primarily interested in trying to improve physical performance and wrestling technique and are not engaged in severe, acute weight loss practices commonly employed before competition. METHODS: This double-blinded, randomized placebo-controlled study involved 20 wrestlers from the University of Oklahoma assigned to either a treatment group (Cr+3; N = 7; 20.4 yr +/- 0.1) receiving 200 micrograms chromium picolinate daily, a placebo group (P; N = 7; 19.9 yr +/- 0.2), or a control group (C; N = 6; 20.2 yr +/- 0.1) using a stratified random sampling technique based on weight classification. Body composition, neuromuscular performance, metabolic performance, and serum insulin and glucose were measured before and immediately following the supplementation and training period. RESULTS: Repeated measures ANOVA indicated no significant changes in body composition for any of the groups. Aerobic power increased significantly (P < 0.002) in all groups, independent of supplementation. There were significant trial and group x trial interactions for upper body endurance (P = 0.038) and relative bench press power (P = 0.050). Post hoc analyses revealed that the C group increased upper body endurance (P = 0.006), but none of the pre- to post-test changes in bench press power were significant. CONCLUSIONS: These results suggest that chromium picolinate supplementation coupled with a typical preseason training program does not enhance body composition or performance variables beyond improvements seen with training alone. PMID- 9861609 TI - A new reliable laboratory test of endurance performance for road cyclists. AB - PURPOSE: The purpose of this study was to devise and evaluate a laboratory test of cycling performance that simulates the variable power demands of competitive road racing. The test is a 100-km time trial interspersed with four 1-km and four 4-km sprints. METHODS: On three occasions separated by 5-7 d, eight endurance trained cyclists (peak oxygen uptake 5.0 +/- 0.7 L.min-1, peak power output 411 +/- 43 W, mean +/- SD) performed the test on their own bikes mounted on an air braked Kingcycle ergometer. Subjects were free to regulate their power output but were asked to complete each sprint and the full distance as quickly as possible. The only feedback given to the cyclists during each test was elapsed distance. RESULTS: In the first test, time for the 100 km and mean times for the 1-km and 4 km sprints were 151:42 +/- 10:36, 1:16 +/- 0:06, and 5:31 +/- 0:16 min:s, respectively; these times improved by 1.6-2.2% in the second test, but there was little further improvement in the third test (0.7 to -0.5%). The between-test correlation for 100-km time was 0.93 (95% CI 0.79 to 0.98), and the within cyclist coefficient of variation was 1.7% (95% CI 1.1 to 2.5%). Mean sprint performance showed similar good reliability (within-subject variation and correlations for the 1-km and 4-km sprint times of 1.9%, 2.0%, 0.93, and 0.81, respectively). CONCLUSIONS: The high reliability of this laboratory test will make the test useful for research on performance of competitive road cyclists. PMID- 9861610 TI - The Cartesian clock metaphor for pineal gland operation pervades the origin of modern chronobiology. AB - In theoretical descriptions formulated during the 1600s, R. Descartes attributed a clock-like role to the pineal gland. He established the belief that pineal function underlies the laws of the universe that determine the cyclic sleep-awake states in man. Recent reports about pineal circadian pacemakers now validate the brilliant accuracy of Cartesian thought, in relation to the relevant role of the pineal gland. PMID- 9861611 TI - Amiloride and vertebrate gustatory responses to NaCl. AB - Amiloride at < or = 1 microM may block epithelial Na+ channels without affecting other cellular mechanisms, and attenuates gustatory responses to lingual NaCl from the chorda tympani nerves (CT) of gerbil, hamster, rhesus monkey, and several strains of laboratory rat and mouse, and from glossopharyngeally innervated frog taste-receptor cells; at 5 microM to 50 microM, also from Wistar rat and mongrel dog CT. Affected units responded more to NaCl than to KCl. Suppression of CT responses to KCl, HCl, NH4Cl, or saccharides also occurred in some mammals, but amiloride did not elicit responses. Taste-dependent behaviors towards NaCl or KCl were altered. DBA and 129/J laboratory mice, and mudpuppy, were unaffected by amiloride. In humans, 10 microM amiloride both produced taste reports and reduced total intensity of NaCl and LiCl by 15-20%. NaCl and LiCl sourness, and KCl and QHCl bitterness declined, but saltiness generally did not change. Effects on sweetness were inconsistent. Amiloride-sensitive gustatory mechanisms were prominent in some mammals, were not necessary for responses to NaCl, and were of minor importance for human taste. PMID- 9861612 TI - Brain mechanisms of mammalian fluid homeostasis: insights from use of immediate early gene mapping. AB - A comprehensive review of the literature through mid-1997 is presented on the application of immediate early gene mapping to problems related to brain mechanisms of fluid homeostasis and cardiovascular regulation in mammals. First, the basic mechanisms of fluid intake and the principles and pitfalls of immediate early gene mapping are briefly introduced. Then, data from several principal paradigms are reviewed. These include fluid deprivation and intracellular dehydration, both of which are associated with thirst and water intake. The contributions of peripheral sodium receptors, and of both hindbrain and forebrain integrative mechanisms are evaluated. Extracellular dehydration, and associated aspects of both thirst and sodium appetite are then reviewed. The contributions of both structures along the lamina terminalis and the hypothalamic magnocellular neurosecretory groups figure prominently in most of these paradigms. Effects of hypotension and hypertension are discussed, including data from the endogenous generation and the exogenous application of angiotensin II. Lastly, we summarize the contribution of the early gene mapping technique and consider briefly the prospects for new advances using this method. PMID- 9861613 TI - Ventrolateral medullary control of cardiovascular activity during muscle contraction. AB - An overview of the role of ventrolateral medulla (VLM) in regulation of cardiovascular activity is presented. A summary of VLM anatomy and its functional relation to other areas in the central nervous system is described. Over the past few years, various studies have investigated the VLM and its involvement in cardiovascular regulation during static muscle contraction, a type of static exercise as seen, for example, during knee extension or hand-grip exercise. Understanding the neural mechanisms that are responsible for regulation of cardiovascular activity during static muscle contraction is of particular interest since it helps understand circulatory adjustments in response to an increase in physical activity. This review surveys the role of several receptors and neurotransmitters in the VLM that are associated with changes in mean arterial pressure and heart rate during static muscle contraction in anesthetized animals. Possible mechanisms in the VLM that modulate cardiovascular changes during static muscle contraction are summarized and discussed. Localized administration of an excitatory amino-acid antagonist into the rostral portion of the VLM (RVLM) attenuates increases in blood pressure and heart rate during static muscle contraction, whereas its administration into the caudal part of the VLM (CVLM) augments these responses. Opioid or 5-HT1A receptor stimulation in the RVLM, but not in the CVLM, attenuates cardiovascular responses to muscle contraction. Furthermore, intravenous, intracerebroventricular or intracisternal injection of an alpha 2-adrenoceptor agonist or a cholinesterase inhibitor attenuates increases in blood pressure and heart rate during static muscle contraction. Finally, the possible involvement of endogenous neurotransmitters in the RVLM and the CVLM associated with cardiovascular responses during static muscle contraction is discussed. An overview of the role of the VLM in the overall cardiovascular control network in the brain is presented and critically reviewed. PMID- 9861614 TI - Play fighting of rats in comparative perspective: a schema for neurobehavioral analyses. AB - Play fighting is a commonly reported form of play in the young of many mammals. Most of the studies on the neurobehavioral mechanisms regulating this behavior have focused on the laboratory rat. The rationale for doing so has been primarily on practical grounds. This paper seeks to answer the question. "How good is the rat as a model of mammalian play fighting?" A review of the detailed structure of play fighting in rats and other mammals reveals that play fighting is not a unitary activity, but rather has distinct components with each having distinct regulatory mechanisms. The rat is typical of many other mammals for some features of play fighting, but not others. Therefore, two conclusions are drawn from this review. First, given that play fighting is a composite category of behavior, questions regarding its underlying neurobehavioral mechanisms need to be narrowly constructed, so as to deal with highly specific mechanisms. For example, what mechanism regulates the pubertal decline in play fighting? Second, the rat is shown to be a good model species for the study of some features of play fighting, but it cannot be assumed to represent an "average" mammal for all features. PMID- 9861615 TI - Potential mechanisms for functional changes in taste receptor cells following sodium deficiency in mammals. AB - Sodium is an essential nutrient for life, and its level in the body is tightly regulated. When sodium deficient, some mammals alter their behavior towards salt by avidly consuming it, even at concentrations animals typically choose to avoid. This change in acceptance is accompanied by a reduction in the response of the gustatory chorda tympani nerve to sodium solutions. More specifically, the response rate of the sodium-specialist units to NaCl stimulation is reduced following sodium deficiency or adrenalectomy. The initial transduction of the chemical signal is mediated, in part, by Na+ influx through epithelial Na+ channels in the apical membrane of taste cells that synapse with the specialist neurons. Circulating hormones like angiotensin II and adrenocorticotropin hormone, which are released in response to sodium deficiency and adrenalectomy, could regulate the activity of Na+ channels through G-protein linked second messenger systems. These putative pathways are of interest because they have been described in mammalian taste receptor cells. The present review will summarize evidence linking some hormones of fluid homeostasis with the apparent attenuation of input from sodium-specialist neurons. PMID- 9861616 TI - Correlation between autotomy-behavior and current theories of neuropathic pain. AB - The past 10 years have brought several new experimental models with which to study chronic neuropathic pain in animals. Consequently, our knowledge about the mechanisms subserving neuropathic pain in humans has improved. However, the first animal model that was used for studying this type of chronic pain was the autotomy-model which can still be considered as a useful tool for pain studies. The present review assesses some of the similarities and differences between autotomy-model and more recent models of experimental traumatic mononeuropathy. In addition, it considers some of the similarities between the results obtained in clinical studies and in autotomy studies. PMID- 9861617 TI - Adverse psychological impact, glutamatergic dysfunction, and risk factors for Alzheimer's disease. AB - Alzheimer's disease (AD) is a neurodegenerative disorder characterized by cell loss and pathological changes in neuronal transmission. In particular, malfunction in glutamatergic activity may be associated with the impairment of memory seen in Alzheimer patients. Both hypoactivation and hyperactivation of glutamatergic systems seem to cause impeded cognitive processing in animals. Rats subjected to rearing in isolation display reduced levels of glutamate in temporal regions accompanied by impaired learning and memory. Similar cognitive deficits are also seen in animals exposed to behavioral stress. Stress appears to have deleterious effects on cognition caused by glutamate neurotoxicity leading to attenuated synaptic activity. It is suggested that stress may represent a potential risk factor for AD. The known risk factors for AD (age, heredity, head trauma, low education, depression) may all be related to glutamatergic dysfunction. Some difficulties with pharmacological approaches based on glutamatergic agonists are discussed. It is suggested that optimal glutamate mediated neurotransmission throughout life may prevent the occurrence of mental decline associated with AD. PMID- 9861618 TI - 13C-urea versus 14C-urea breath test: is there still a need for 14C-urea? PMID- 9861619 TI - An update of radiopharmaceutical schedules in children. AB - Proposed radiopharmaceutical schedules based on readily measured parameters (age, height, weight, surface area) have been considered with respect to their applicability in nuclear medicine. Although schedules based on age are considered to be inappropriate, there are valid cases for schedules based on height, weight and surface area. For many radiopharmaceutical studies, the most appropriate schedules are based on weight or surface area. Examination of simple theoretical physical models, including the influence of attenuation, suggests that the optimum administered amounts of radiopharmaceuticals fall mainly within the region bounded by these two schedules. The results of recent clinical studies designed to test the validity of different schedules are summarized. In general, they support the predictions of theoretical models, but also show how simple models can be influenced significantly by the age dependency of radiopharmaceutical biodistribution. The schedule based on surface area (or height) is less likely than that based on weight to require the identification of minimum administered amounts of radiopharmaceutical to preserve image quality in small children. However, recent studies have shown that the most appropriate schedule for regional cerebral blood flow with 99Tcm-HMPAO is that based on weight, without the need for a minimum activity. Paediatric radiation dosimetry is briefly summarized to indicate some recent innovations in methodology. Effective doses per unit of administered radiopharmaceutical (mSv MBq-1) have been calculated using five paediatric phantoms for a number of radiopharmaceuticals commonly used in children. Values of total effective dose resulting from the application of the weight and surface area schedules are presented, based on the adult reference amounts of administered radiopharmaceutical proposed by the Paediatric Task Group of the European Association of Nuclear Medicine. Although some values of effective dose exceed 10 mSv for the surface area schedule, the majority of values are less than 5 mSv. PMID- 9861620 TI - Immunoscintigraphy (BW 250/183) in neonates and infants with fever of unknown origin. AB - Fever of unknown origin is defined as a temperature above 39.0 degrees C together with a white blood cell count > or = 15,000 mm-3, the duration of fever exceeding 2 weeks and a correct diagnosis not being obtained in the first week of hospitalization. In neonates and infants with fever of unknown origin, the localization of the infectious focus is often difficult and unsatisfactory. In this retrospective study, the clinical value of 99Tcm-labelled antigranulocyte antibodies for this group of patients was investigated. Thirty-two immunoscintigrams were performed using 185-259 MBq 99Tcm-labelled antigranulocyte antibodies (BW 250/183) in 30 neonates and infants (21 boys, 9 girls, mean age 29.4 +/- 2 months), who had fever of unknown origin. Immunoscintigraphy was carried out as whole-body images (n = 7) or single planar images (n = 25) 4 h and 24 h post-injection. In children with known cardiac failure, single photon emission tomography of the thorax was performed to diagnose endocarditis (n = 2). For verification, the results of the immunoscintigrams were compared with radiology (conventional radiography = 14, MRI = 5, CT = 3), biopsy (n = 2), blood culture (n = 10) and clinical follow-up after specific therapy. In 11 of 30 children (36%), the diagnosis of an infective focus was possible with immunoscintigraphy. The sensitivity and specificity of diagnosing infective foci was 72% and 95% respectively (n = 11; colitis = 2, infection of the central permanent catheter tip = 2, middle ear infection = 1, spondylitis/discitis = 3, osteomyelitis = 2, umbilical infection = 1). In vertebral body infections, all lesions were photopenic. In 18 children (60%), no infective focus was found on immunoscintigraphy. In this group of children, the main reason (n = 5) for fever of unknown origin was chronic juvenile rheumatoid arthritis. No uptake was seen in two infants with cardiac failure and suspected endocarditis on SPET. In 3 of the 18 patients (17%), localization of an infective focus was not possible with immunoscintigraphy or on other examinations. In these patients, the fever disappeared spontaneously after a few days of antibiotic therapy. In conclusion, we have shown that 99Tcm-anti-NCA-95 scanning is a safe method with a high sensitivity and specificity for detecting infectious foci in neonates and infants with fever of unknown origin. Furthermore, this method is easy to perform, since no withdrawal of blood is necessary. PMID- 9861621 TI - Can dual-headed 18F-FDG SPET imaging reliably supersede PET in clinical oncology? A comparative study in lung and gastrointestinal tract cancer. AB - In this study, we prospectively compared the sensitivity of PET and planar SPET (collimated gamma camera) 18F-FDG imaging in patients with lung and gastrointestinal tract cancer and analysed their respective impact on patient management. Twenty-eight patients with lung cancer and 14 with gastro-intestinal tract tumours were scanned on the same day with a PET and a collimated planar SPET gamma camera. The planar SPET procedure consisted of whole-body planar views and a tomographic acquisition centred over the torso or the abdomen, with the total imaging time within the same range as the whole-body PET procedure. The staging of lung cancer patients was accurate in 86% with PET and 64% with planar SPET. Planar SPET would have led to inappropriate therapeutic decisions in 8 of 28 patients, mainly due to undetected distant metastases. In patients with suspected gastrointestinal tract cancer, planar SPET identified 7 of 15 (47%) proven tumour sites, whereas PET identified 14 of 15 (93%). Our results suggest that collimated planar SPET cameras are not a substitute for dedicated PET scanners. The sensitivity for the detection of tumours is unacceptably low and can impair patient management. The use of multiple tomographic acquisitions could improve the sensitivity but would require a longer scanning time. PMID- 9861622 TI - Positron emission tomography with 18F-FDG to detect residual disease after therapy for malignant lymphoma. AB - We retrospectively evaluated the use of 18F-FDG PET for assessment of residual disease in 27 patients after therapy for malignant lymphoma. The images were evaluated qualitatively and quantitatively using standardized uptake values (SUV). All findings were validated either by biopsy or by clinical follow-up and compared with corresponding CT findings. The impact of blood glucose concentration, body weight, body surface area, lesion diameter and the time between injection and imaging on the SUVs were analysed. All 15 patients with biopsy-proven residual disease or relapse during follow-up and 11 of 12 patients who remained relapse-free were correctly identified by qualitative interpretation of the PET images. A case of pneumonitis after radiotherapy/chemotherapy accounted for the only false-positive finding. Compared with CT imaging, PET had a significantly higher specificity (P < 0.01), accuracy (P < 0.05) and positive predictive value (P < 0.05). The mean and maximum SUV of the tumour lesions were positively correlated to lesion diameter (P < 0.01) and imaging time post injection (P < 0.01). Standardized uptake values corrected for the partial volume effect and normalized to a standardized imaging time (SUVBPT) were significantly higher (P < 0.05) in high-grade than in low-grade non-Hodgkin's lymphoma. In conclusion, 18F-FDG PET may help in the identification of patients who need additional treatment after the completion of conventional therapy. Qualitative image interpretation appears sufficient for this purpose. PMID- 9861623 TI - Comparison of SPET brain perfusion and 18F-FDG brain metabolism in patients with chronic fatigue syndrome. AB - Chronic fatigue syndrome is a clinically defined condition of uncertain aetiology. We compared 99Tcm-HMPAO single photon emission tomography (SPET) brain perfusion with dual-head 18F-FDG brain metabolism in patients with chronic fatigue syndrome. Eighteen patients (14 females, 4 males), who fulfilled the diagnostic criteria of the Centers for Disease Control for chronic fatigue syndrome, were investigated. Thirteen patients had abnormal SPET brain perfusion scans and five had normal scans. Fifteen patients had normal glucose brain metabolism scans and three had abnormal scans. We conclude that, in chronic fatigue syndrome patients, there is discordance between SPET brain perfusion and 18F-FDG brain uptake. It is possible to have brain perfusion abnormalities without corresponding changes in glucose uptake. PMID- 9861624 TI - The discrepancy between 99Tcm-ECD dynamic and static SPET images in patients with ischaemic lesions corresponds to reduced vasoreactivity to acetazolamide. AB - We evaluated 99Tcm-N,N'-(1,2-ethylenediyl)bis-L-cysteine diethyl ester (99Tcm ECD) dynamic and static SPET (single photon emission tomographic) images to examine 99Tcm-ECD kinetics under ischaemic cerebrovascular conditions. In 20 patients who showed arterial occlusion on magnetic resonance angiography, dynamic (0-10 min) and static (15-35 min) SPET images were acquired after the intravenous administration of 99Tcm-ECD. Thirteen of the patients had focal perfusion deficits that were more evident on the dynamic than on the static images; the other seven showed no such discrepancy. In those patients with a mismatch between the dynamic and static images, the extent corresponded to reduced vaso-reactivity to acetazolamide. Based on quantitative analysis of the ratio of tracer uptake in affected to that in unaffected areas, the patients with discrepant findings showed significantly different ratios on the dynamic and static images, whereas those with no such mismatch did not. Our results suggest that dynamic 99Tcm-ECD images provide circulatory information and that static images reflect a filling in phenomenon of ECD metabolites in ischaemic lesions. 99Tcm-ECD dynamic and static SPET images offer an alternative method of detecting mild perfusion deficits without the need for acetazolamide challenge. PMID- 9861625 TI - Extent of myocardial damage in regions with reverse redistribution at 3 h and at 24 h on 201Tl SPET: evaluation based on regional myocardial oxidative metabolism. AB - Reverse redistribution (RRD) of 201Tl is often observed in patients with recent myocardial infarction. However, the difference in the extent of myocardial damage between regions with 3-h RRD and those with 24-h RRD remains unknown. Accordingly, we investigated RRD from the standpoint of myocardial oxidative metabolism. Carbon-11 (11C) acetate dynamic myocardial PET scanning was performed at rest in 14 patients with recent myocardial infarction, and the clearance rate constant (Kmono) of 11C-acetate was calculated in 6-7 ROIs on the transaxial image in each patient using a monoexponential fit as an index of myocardial oxidative metabolism. Exercise 201Tl myocardial SPET was also performed. Ninety two regions corresponding to the PET study were then classified based on the findings of transaxial 201Tl SPET imaging; that is, regions with reverse redistribution, regions with severely decreased 201Tl activity or no 201Tl activity on the 24-h delayed images, and regions with normal 201Tl activity throughout the study. Kmono in regions with reverse redistribution (0.051 +/- 0.009 min-1) was significantly lower than that in regions with normal 201Tl activity throughout the study (0.066 +/- 0.011 min-1) (P < 0.001) but significantly higher than that in regions with severely decreased or no 201Tl activity on the 24-h delayed images (0.037 +/- 0.003 min-1) (P < 0.001). Percent Kmono (i.e. Kmono in region with RRD/the mean of Kmono in all regions with a normal 201Tl SPET result) was significantly lower in the 3-h RRD regions (81.3 +/ 6.3%) than in the 24-h RRD regions (87.6 +/- 6.1%) (P < 0.05). Impairment of myocardial oxidative metabolism is observed in regions with RRD, suggesting that RRD corresponds to mild myocardial damage. Reverse redistribution on 24-h delayed images may indicate much milder myocardial damage compared with RRD on 3-h delayed images. PMID- 9861626 TI - Quantification of gamma camera spatial resolution by means of bar phantom images. AB - A method that has previously been described for rapid objective measurement of intrinsic gamma camera spatial resolution using statistical moments has been applied to images of a four-quadrant bar phantom acquired with various collimators. The full-width at half-maximum (FWHM) of the line spread function (LSF) and modulation transfer functions determined by the moments method was compared with those obtained directly from line spread functions. It was found that, for highest accuracy, the formulae originally described for intrinsic measurements have to be modified to take account of a more accurate description of the Fourier components of the input from a bar phantom. When this modification was applied, FWHM measurements agreed to within 4.0% with the LSF measurements in the range 4.5-7.2 mm if well-resolved images of bars wider than 3.0 mm were used. The method is simple to apply and since the coefficient of variation of these measurements was < 2.2%, it is well-suited for rapid objective routine monitoring of gamma camera spatial resolution. For modulation transfer functions > 0.2, bar sizes of 4.0 and 4.5 mm gave values within 4% of the LSF measurements, when the modified formula was applied, with coefficients of variation less than 4%. PMID- 9861627 TI - Effect of nifedipine and verapamil on hypotensive action of clonidine in rabbits. AB - The present study examined the influence of nifedipine and verapamil on clonidine induced hypotension in the rabbits. Clonidine was given intravenously (i.v.) (0.2 mg/kg) or intracerebroventricularly (icv) (0.03 mg). Nifedipine (0.3 mg/kg and 0.45 mg/kg) and verapamil (0.2 mg/kg and 0.3 mg/kg) were injected i.v. 15 min before clonidine administration or icv (nifedipine 0.03 mg, verapamil 0.1 mg) 10 min before clonidine. It has been shown that nifedipine administered i.v. increases whereas injected icv prevents the hypotensive action of clonidine administered in the same way. Verapamil injected i.v. and icv did not change the hypotensive action of clonidine. It seems that nifedipine has alpha 2 adrenoceptor blocking properties. PMID- 9861628 TI - Interaction of cholecystokinin (CCK-33) and its C-terminal fragments: CCK-8 and CCK-4 with alpha- and beta-adrenoceptor agonists and antagonists in the cardiovascular system of rats. Part A. AB - The goal of the present study was to examine the interaction of cholecystokinin (CCK-33) and its fragments: C-terminal octapeptide (CCK-8) and C-terminal tetrapeptide (CCK-4) with alpha- and beta-adrenoceptor agonists and antagonists. The effect of this interaction on arterial blood pressure and function of isolated heart was studied in rats. The results indicate that: 1) CCK-33 enhances the influence of catecholamines: noradrenaline and isoprenaline, mainly on the function of isolated heart. This peptide does not change cardiovascular effects of alpha-adrenoceptor antagonist--phentolamine. CCK-33 diminishes the influence of propranolol on the function of isolated heart. The hypotensive effect of beta adrenoceptor antagonist is not affected by CCK-33. 2) CCK-8 does not alter cardiovascular effects of noradrenaline and isoprenaline. The peptide diminishes the hypotensive effect of phentolamine and reverses the hypotensive effect of propranolol. CCK-8 enhances the influence of propranolol and does not change the influence of phentolamine on the function of isolated heart. CCK-8 enhances bradycardia evoked by propranolol. 3) CCK-4 does not change the influence of noradrenaline and isoprenaline on arterial blood pressure and diminishes the hypotensive effect of phentolamine and propranolol. The peptide does not change cardiac effects of noradrenaline and diminishes the effects of isoprenaline, phentolamine and propranolol. On the basis of the present study, we concluded that CCK-33 and its fragments CCK-8 and CCK-4 modify the cardiovascular action of alpha- and beta-adrenoceptor agonists and antagonists. We suggest that effects we have observed correlate with the activation of the CCK-A receptors (CCK-33, CCK 8) or CCK-B receptors (CCK-4). CCK-related peptides may increase or reduce the effects of catecholamines indirectly through activation of alpha-adrenoceptors. We can not exclude direct action of the peptides on the heart. PMID- 9861630 TI - Evaluation of antiarrhythmic activity of captopril and enalaprilat in experimental cardiac arrhythmias in rabbits. Part I. AB - The present study was undertaken to investigate the influence of captopril and enalaprilat--two different angiotensin converting enzyme (ACE) inhibitors--on experimental cardiac arrhythmias induced by barium chloride, ouabain or adrenaline. The research was carried out on rabbits. Captopril and enalaprilat were administered only once. Arrhythmias were evoked by three different doses of arrhythmogens: ED50 and two higher ones causing rhythm disturbances in 80-100% rabbits. The patterns of arrhythmias as well as their frequency and duration were evaluated on the basis of ECG examination. The results were subjected to statistic analysis. As a result of our research, we were able to establish that ACE inhibitors, when administered at a single dose, did not influence the patterns but changed the frequency and duration of barium chloride-, adrenaline- or ouabain-induced arrhythmias. Captopril and enalaprilat administered at a single dose decreased the frequency of barium chloride- or ouabain- but not adrenaline-induced arrhythmia. A single administration of captopril and enalaprilat limit the duration of arrhythmias caused by barium chloride, ouabain or adrenaline. PMID- 9861629 TI - Interaction of cholecystokinin (CCK-33) and its C-terminal fragments: CCK-8 and CCK-4 with alpha- and beta-adrenoceptor agonists and antagonists in the cardiovascular system of diabetic rats. Part B. AB - This study investigated the interaction of cholecystokinin (CCK-33) and its fragments: C-terminal octapeptide (CCK-8) and C-terminal tetrapeptide (CCK-4) with alpha- and beta-adrenoceptor agonists and antagonists. The effects of this interaction on arterial blood pressure and function of isolated heart were studied in rats with 1-month diabetes mellitus (DM) induced by 1-month administration of streptozocin (STZ). We used CCK-33, CCK-8 and CCK-4 in an equimolar dose of 425.0 pmoles/kg i.v. in in vivo, and 42.5 pmoles/0.1 ml in in vitro experiments. We found that 1) DM diminished the hypertensive effect of noradrenaline (NA) and the hypotensive effects of isoprenaline (ISO), phentolamine (PHENT) and propranolol (PROP). CCK-4 and CCK-8 enhanced but CCK-33 did not change a weaker hypertensive effect of NA while all peptides showed the hypotensive effect of PHENT and did not change the hypotensive effect of ISO and PROP. 2) The influence of NA and PROP on isolated heart of diabetic rats remained unchanged after administration of the peptides but that of ISO and PHENT was diminished. CCK-4 enhanced the effects of NA and PHENT, and diminished the influence of ISO and PROP on isolated heart of diabetic rats. CCK-33 enhanced the cardiac effects of NA, ISO and PROP and did not alter PHENT action. CCK-8 did not change the influence of the studied adrenoceptor agonists and antagonists on isolated heart of diabetic rats. The results of the present study demonstrated that the smallest fragment of CCK-33 (CCK-4) modified the influence of alpha- and beta-adrenoceptor agonists and antagonists on cardiovascular system and altered the direction of disturbances caused by DM. The other peptides (CCK-33, CCK-8) interacted with these receptors to a lesser degree in diabetic subjects. PMID- 9861631 TI - Evaluation of antiarrhythmic activity of captopril and enalaprilat in experimental cardiac arrhythmias in rabbits. Part II. AB - The influence of 21-day administration of captopril and enalaprilat on barium chloride and adrenaline-induced experimental arrhythmias was assessed. The experiments were performed on rabbits. Arrhythmias were evoked by two alternative arrhythmogen doses. The patterns of disturbances, their frequency and duration were evaluated on the basis of ECG examination. Antiarrhythmic properties of angiotensin converting enzyme inhibitors administered for 21 days were also compared with their effects after single administration. The results were subjected to statistic analysis. On the basis of the obtained results we were able to establish that repeated administration of enalaprilat decreases the frequency of barium chloride- and adrenaline-induced arrhythmias. Repeated administration of captopril and enalaprilat shortened the duration of adrenaline- and barium chloride-induced arrhythmias. Long-term enalaprilat administration was much more effective in preventing arrhythmias than its single dose, it also proved to be more efficient than either single or repeated administration of captopril. PMID- 9861632 TI - Ecto-ATPase from rat lymphocytes--in vivo studies on the influence of levamisole. AB - Levamisole, known for a long time as an antiparasitic drug due to its immunomodulating properties, has been used in the therapy of immune disorders. The action of levamisole depends on the dose and the manner of administration, and it seems interesting to establish whether similar dependencies exist for the membrane enzymes of lymphocytes. Ecto-ATPase, the enzyme tightly integrated with cytoplasmic membrane, was chosen to examine these effects. The in vivo studies were conducted on rats with carragenine-induced inflammation. Levamisole was administered at two doses: high (2.5 mg/kg) and low (0.25 mg/kg), according to two schedules: a) at a single dose, simultaneously with carragenine, 24 h before or 24 h after induction of inflammation; the lymphocytes were collected five times every 24 h, and the activity of ecto-ATPase was determined; b) at four doses, every 24 h after carragenine injection; the lymphocytes were collected 120 h after the induction of inflammation. The results of our studies indicate that levamisole reduces the activity of ecto-ATPase in both populations of lymphocytes in vivo. An apparent deactivation of ecto-ATPase of lymphocytes T was observed when high doses of levamisole were administered four times. In the case of a single low dose of levamisole, the deactivating potential of this compound was not so evident, both for lymphocytes B and T. Different reactions of the enzyme from lymphocytes T and B to the levamisole observed in in vivo studies and different effects of inhibitors in vitro, suggest the dissimilarity of the structure of ecto-ATPases from both populations of lymphocytes. PMID- 9861633 TI - Effects of pamidronate on the development of changes in bone mechanical properties and bone structure caused by the administration of prednisolone in rats. AB - The purpose of the study was to investigate the influence of pamidronate on mechanical properties, growth, and structural changes in bones of rats in which experimental osteopenia was induced by administration of prednisolone. The experiment was carried out on male WAG rats divided into three groups: I. Control, II. Prednisolone (5 mg/kg im daily) and III. Disodium pamidronate (3 mg/kg sc daily) + prednisolone (5 mg/kg im daily). After three weeks of the experiment, the animals were sacrificed and their femoral and tibial bones were prepared. The administration of prednisolone resulted in morphological and metabolic changes in the osseous system, characteristic of osteopenia. The increased osteopsathyrosis was noted, manifesting in lowered resistance to fractures and lesser deformability in comparison with the control group. The administration of pamidronate resulted in the reduction of the destructive action of prednisolone on bones. PMID- 9861634 TI - Effect of kainic acid and pentetrazole administration on the activity of nitric oxide synthase in the mouse brain. AB - The present study examined ex vivo effect of kainic acid and pentetrazole administration on the activity of nitric oxide synthase (NOS) in the mouse brain. NOS activity was assayed by measuring the formation of [3H] citrulline from [3H]arginine in the homogenates of mouse hippocampus, neocortex and cerebellum. The highest basal activity of the enzyme was found in this latter brain region. Administration of kainic acid (30 mg/kg) increased the NOS activity in all brain regions examined. On the other hand, pentetrazole (60 mg/kg) did not evoke any significant changes in the NOS activity at 5 min after the administration. Only in cerebellum, at 10 min after administration of pentetrazole, the increase in the activity of the enzyme was observed. The obtained results indicate that the two particular convulsants used in this study differ not only in respect of behavioral signs of seizures which they evoke, but also in respect of the effect on mouse brain NOS activity. PMID- 9861635 TI - BN 52021, PAF-receptor antagonist, improves diminished antioxidant defense system of lungs in experimentally induced haemorrhagic shock. AB - The purpose of our experiments was to determine whether BN 52021, PAF-receptor antagonist, has any influence on the production of reactive oxygen species in the lung induced by haemorrhagic shock. The results showed, that the activities of Cu,Zn-superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the lung after haemorrhagic shock were decreased in comparison with control and sham groups while the activity of glutathione reductase (GSSG-R) was unchanged. The content of SH-groups was decreased while thiobarbituric acid reacting substances (TBA-rs) after haemorrhagic shock were increased. Infusion of BN 52021 (5.0 mg/kg i.v.) in haemorrhagic animals caused significantly increased activities of Cu,Zn SOD, GSH-Px, GSSG-R in the lung. The SH-group level was slightly increased while TBA-rs returned to normal values. PMID- 9861636 TI - Morphine reverses the inhibitory effects of repeated saline injections on peritoneal inflammation in mice. AB - Morphine sc injections reversed the inhibitory effects of daily sc saline injections on the thioglycollate-elicited peritoneal inflammation in Swiss mice. PMID- 9861637 TI - European Collaborative Project on Affective Disorders: interactions between genetic and psychosocial vulnerability factors. AB - Despite strong evidence provided by genetic epidemiology of genetic involvement in the aetiology of bipolar and unipolar affective disorders, the exact nature of the predisposing gene(s) is still being investigated through linkage and association studies. The interaction of susceptibility genes and environmental factors in these diseases is also of fundamental importance and requires proper investigation. Interesting theories have recently been proposed examining the possible role of various chromosomal regions, candidate genes and mutations in affective disorders. Reliable multicentre-based methodology is currently being employed to examine these theories, with attention given to statistical analysis and the statistical power of the sample. The present article describes the European Collaborative Project on Affective Disorders (ECPAD) 'Interactions between genetic and psychosocial vulnerability factors', involving 15 European centres. A description is given of the association and family samples collected for the project and also the methodology used to analyse interactions in the gene psychosocial environment. This material provides a powerful tool in the search for susceptibility genes in affective disorders and takes into account non genetic aetiological factors. PMID- 9861638 TI - Schizophrenia and the serotonin transporter gene. AB - A case control study was conducted among cases with schizophrenia (DSM IV criteria) and screened adult controls from three cohorts. Bi-allelic polymorphisms in the promoter region of the serotonin transporter gene (5-HTT) were examined in conjunction with those of the serotonin 5-HT2a receptor (HTR2). No significant association with 5-HTT was detected among US Caucasians (n = 207), African-Americans (n = 84) or Caucasians from Sweden (n = 221). However, survival analysis suggested an association with the age at onset among the Swedish cases. The association should be considered tentative as it was not evident in the smaller US samples. The following exploratory analyses among the US samples were also not significant: associations with subgroups of patients based on familiality or response to medications, or altered risk due to the joint effects of 5-HTT and HTR2 genotypes. PMID- 9861639 TI - Parent-of-origin effect and evidence for differential transmission in periodic catatonia. AB - In a family study involving 83 probands with periodic catatonia a subtype of DSM IIIR schizophrenia, we reported an age-specific morbidity risk of 26.9% in first degree relatives with homotypical psychoses and genetic anticipation indicating a possible major gene effect. Paternal transmission was associated with a trend for a younger age at onset in probands compared to that observed in the case of maternal transmission (P = 0.099). If this can be confirmed in a larger sample and then replicated, there would be evidence for the occurrence of a parent-of origin effect. Such an observation may indicate that a paternally imprinted locus acts on periodic catatonia. Among the non-genetic mechanisms that may modify the penetrance of the disease, paternal affection did lead to a decrease in male offspring (P = 0.007) and maternal affection showed an increased frequency of non affected male offspring (P = 0.021). We therefore propose that parent-of-origin effects as well as prenatal mortality and psychosocial factors need further investigation in the periodic catatonia subtype of schizophrenia. PMID- 9861640 TI - Preliminary evidence of an association between bipolar disorder in females and the catechol-O-methyltransferase gene. AB - Catechol-O-methyltransferase (COMT) catalyses the methylation, and hence the inactivation, of catecholamines including the neurotransmitters dopamine and noradrenaline. There is evidence implicating COMT as a candidate gene for a number of neuropsychiatric conditions including bipolar disorder. A long recognized population variation in COMT activity exists and it has recently been established that variation in enzyme activity results from a polymorphic genetic variation within the COMT gene which can be readily assayed as a polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP). A collection of 60 Irish bipolar I probands have been genotyped together with their parents. Tests comparing transmitted and non-transmitted alleles provide no evidence that the polymorphism contributes to a susceptibility to bipolar disorder within the sample as a whole. However, amongst female bipolar I probands (n = 30) there was a tendency for the low-activity allele of COMT to be preferentially transmitted. Furthermore, a re-examination of an Irish case control sample resulted in a similar observation amongst female bipolar I sufferers and pooling the data sets strengthened the findings. PMID- 9861641 TI - Genotyping microsatellite polymorphisms by agarose gel electrophoresis with ethidium bromide staining: application to quantitative trait loci analysis of seizure susceptibility in mice. AB - Agarose gel electrophoresis with ethidium bromide staining (AGE/EBS) is an efficient and reliable method for analyzing microsatellite polymorphisms. We report the use of AGE/EBS for analyzing DNA microsatellite polymorphisms in a preliminary quantitative trait loci (QTL) study of seizure susceptibility in which a candidate gene strategy was used to direct initial mapping efforts. F2 intercross progeny, derived from seizure-sensitive DBA/2J (D2) and seizure resistant C57BL/6J (B6) inbred strains of mice, were tested for their sensitivity to the seizure-inducing effect of pentylenetetrazol (PTZ), a gamma-aminobutyric acid (GABA) receptor antagonist. A semi-automated method is described, in which DNA microsatellites were amplified by polymerase chain reaction (PCR) to yield products of 100-200 base pair (bp) in length. Alleles were separated on 3-6% MetaPhor agarose gels, stained with ethidium bromide, and visualized by ultraviolet (UV) illumination. Univariate analysis of genotype and phenotype data provides evidence for a seizure-related QTL on chromosome 5, near genes coding for the GABAA receptor subunits alpha 5 and gamma 3. Interestingly, this suggestive QTL derives from the more resistant B6 strain, but it nonetheless provides impetus for the characterization of possible strain differences in these two candidate genes. Overall, these results demonstrate that AGE/EBS can be useful for rapid screening of genomic regions of special interest in QTL mapping studies. PMID- 9861643 TI - Search for a schizophrenia susceptibility gene on chromosome 13. PMID- 9861642 TI - Human G(olf) gene polymorphisms and vulnerability to bipolar disorder. AB - Two intronic polymorphisms of the human alpha subunit of the olfactory G-protein (G(olf)) are described. They were detected with single-stranded conformational polymorphism (SSCP) methods and confirmed by sequencing both strands. These single base pair (bp) substitutions occur in introns 3 (an A/G at 35 bp 3' from the exon 3/intron 3 5' splice site) and 10 (an T/G at 7 bp 5' from the 3' splice site). Both polymorphisms are relatively common, with minor allele frequencies of 31% (intron 3) and 16% (intron 10). The intron 3 variant shows no linkage disequilibrium with an intron 5 (CA)n microsatellite located approximately 50 kb 3' from the intron 3 variant, among a small group of German individuals with schizophrenia. The intron 3 variant is interesting because it may create an 'in frame' cryptic splice site which, if activated, would add 12 residues to exon 3. The intron 10 variant is interesting because a purine is substituted for a pyrimidine in the 'polypyrimidine' tract of the 3' splice site, a single base substitution of the type which has been associated with aberrant splicing in the androgen receptor gene. PMID- 9861644 TI - A linkage study of affective disorders in two Bulgarian Gypsy families: results for candidate regions on chromosomes 18 and 21. PMID- 9861645 TI - GABAA alpha-1 subunit gene not associated with depressive symptomatology in mood disorders. AB - Considerable evidence implicates the neurotransmitter gamma-aminobutyric acid (GABA) in the biochemical pathophysiology of mood disorders. In this study, we investigated the possibility that the gene for the gamma-aminobutyric acid type A (GABAA) receptor alpha-1 subunit (GABRA1) might be associated with depressive symptomatology in a sample of mood disorder subjects. Sixty-seven inpatients affected by unipolar (n = 37) and bipolar (n = 30) disorder (DSMIV) were assessed at admission by the Hamilton depression rating scale (HAMD) and were typed using polymerase chain reaction (PCR) techniques. GABRA1 variants were not associated with depressive symptomatology, and consideration of possible stratification effects such as sex, psychiatric diagnosis and illness severity did not reveal any association either. GABAA alpha-1 subunit gene is not, therefore, associated with depressive symptomatology in mood disorder subjects. PMID- 9861646 TI - A higher frequency of a low activity-related allele of the MAO-A gene in females with obsessive-compulsive disorder. PMID- 9861647 TI - Intrafamilial association of pericentric inversion of chromosome 9, inv (9)(p11 q21), and rapid cycling bipolar disorder. AB - Association of a chromosome aberration and psychiatric disorder can be useful in highlighting a genomic region that can be profitably explored further using positional cloning. We report the case of a father and daughter both of whom have bipolar disorder II and a pericentric inversion of chromosome 9. PMID- 9861648 TI - [Classification and grading of gliomas and meningiomas]. AB - GLIOMAS: As we demonstrated for supratentorial, diffuse gliomas in adults, a stratification into just two grades of malignancy, 'low' and 'high grade,' proved reliable and prognostically relevant. The discriminating histomorphological criterion for high-grade astrocytoma (WHO glioblastoma) as well as anaplastic oligodendroglioma and anaplastic oligoastrocytoma is endothelial hyperplasia/proliferation, which is usually associated with uptake of contrast medium in computed tomography and magnetic resonance imaging. As neoangiogenesis indicates glioma progression, it is worthwhile considering these radiographic features to judge the representativeness of the tumor samples critically. MENINGIOMAS: The revised edition of the WHO classification of brain tumors now includes the 'atypical' meningioma (WHO 'grade' II): Based on both its histomorphological features and prognosis, it should be placed between the common type and anaplastic meningioma. Nuclear area related Ki-67 proliferation indices, as determined by morphometry, were the prerequisite for outlining its histomorphological spectrum better. Cytogenetically, the most consistent progression-associated feature was loss of the distal part of the short arm of one chromosome 1 (1p-). Thus, a screening method using the tissue non-specific form of alkaline phosphatase (ALPL) as the respective marker enzyme was established. Diagnosing a meningioma of the intermediate type implies careful clinical and radiological patient follow-ups to detect tumor recurrences early. PMID- 9861649 TI - [Genetic factors in the pathogenesis of brain tumors]. AB - The biological properties of tumor cells are determined by, mostly acquired, alterations of proliferation-relevant genes that can provide valuable diagnostic and prognostic parameters. Meningiomas and gliomas are among the genetically best characterized solid tumors. Here, specific patterns of primary and secondary genetic aberrations have been identified and shown to be correlated with clinical parameters such as recurrency of meningiomas or malignant progression of astrocytomas. Focal progression leading to intratumoral heterogeneity, which is most strikingly seen in gliomas, results from complex mechanisms of mutation and selection of tumor cells that are also iatrogenically influenced through radiotherapy and chemotherapy. PMID- 9861650 TI - [Brain stem glioma]. AB - Brain-stem gliomas occur mainly in childhood and are localized in the mesencephalon, pons and medulla oblongata. Diagnosis is a domain of MRI, requiring T2, T1 and KM. CT shows hemorrhage and calcification well. The criteria are the primary site, size, tumor growth, brain-stem enlargement, delineation, intralesional structure, exophytic components and enhancement. Secondary criteria are herniation, hydrocephalus and liquorgenic seeding. In CT glioma are hypodense, in MRI hyperintense in T2 and hypointense in T1. Enhancement is seen in 25-60% and does not allow differentiation of tumor vs nontumor or gradings. Factors influencing poor outcome are high grade, a short history, cranial nerve involvement, severe brain-stem enlargement, pontine site, diffuse growth and recurrency. The 5-year-survival rate is 30% (after radiation: focal tumors 85%, diffuse 20%). Most frequent are symptoms of brain pressure, cerebellum, cranial nerves and pyramidal tract. There is no agreement on whether biopsy is necessary or not. A diagnosis of tumor is highly suggestive if classical MRI findings fit the clinical history. PMID- 9861651 TI - [Neuroradiologic diagnosis of primary non-Hodgkin's lymphoma of the brain]. AB - The incidence of primary cerebral lymphomas has risen continuously during the past years. The neuroradiological signs, which are decisive for the differential diagnosis of cerebral lymphomas, are worked up and discussed in this study. Thirty CT and 27 MR investigations of a total of 32 patients (14 males, 18 females, age 60 +/- 15 years) with histopathologically proven cerebral lymphomas were analysed retrospectively. Multiple cerebral lymphomas were detected in 10/32 patients (31%). The cerebral lymphomas appeared on CT scans as hyperdense masses in 28/30 cases (93%). On T2-weighted MR scans 14/27 (52%) lymphomas were slightly hyperintense compared to white matter and 9/27 (33%) inhomogeneously isointense to poorly hyperintense. These were clearly T2-hypointense compared to T2 hyperintense perifocal oedema. In these cases CT density and T2-weighted signal intensity looked like grey matter. All cerebral lymphomas except one case took up contrast medium. An ependymal infiltration or a contact to the ventricle's wall were found in 24/32 cases and an infiltration or a contact to the leptomeningeal space in 15/32 cases. One should consider a cerebral lymphoma as a possible differential diagnosis when a cerebral mass shows the following signs: (1) in CT scans as a hyperdense mass, and on T2-weighted MR images hyperintense compared to white matter and hypointense compared to perifocal edema; (2) clear contrast enhancement and (3) infiltration or broad contact with the ependyma and/or the leptomeningeal space. PMID- 9861652 TI - [Contribution of nuclear medicine to the diagnosis of recurrent brain tumors and cerebral radionecrosis]. AB - The evaluation of brain tumor recurrence and therapy-induced benign changes following surgery and/or irradiation is a diagnostic challenge for imaging methods based on either morphology (cCT/MRI) or function (SPECT/PET). Current literature and the present data of our own patients demonstrate the diagnostic efficiency of IMT-SPECT and FDG-PET in the detection of recurrence and in-vivo grading. Thirty-nine patients suspected of brain tumor recurrence at follow-up were studied by FDG-PET and IMT-SPECT. Thirty-four of 39 patients showed recurrences; in 12 cases even a change in the grade of malignancy was observed. All high-grade recurrences could be confirmed by either methods. IMT-SPECT showed a higher sensitivity in detecting low-grade tumors at recurrence. In contrast to IMT-SPECT, FDG-PET supports sufficient in-vivo grading. Both methods can be used to differentiate between tumor recurrence and radionecrosis. In conclusion the results of our study demonstrate the efficiency of IMT-SPECT and FDG-PET in confirming recurrences and determining the actual tumor grade. PMID- 9861653 TI - [Xenon-CT and perfusion MRI in the diagnosis of cerebral gliomas]. AB - Within certain limits the vascularity of cerebral gliomas correlates with dysplasia. Xenon CT and perfusion MRI can be used to investigate tumor vascularity. Using an optimized wash in/out protocol with inhalation of 30% xenon gas, xenon CT was performed and regional cerebral blood flow (rCBF) and blood/tissue partition coefficient (lambda) calculated. Furthermore, perfusion MRI was performed using a dynamic T2*w EPI sequence with intravenous administration of Gd-DTPA. The relative regional cerebral blood volume, arrival time, time to peak and mean transit time were calculated. Both modalities were used in patients with supratentorial gliomas and compared with conventional contrast-enhanced MRI (ceMRI). Calculation of regional cerebral blood flow, as compared to ceMRI, did not contribute to the diagnostic approach of tumor dysplasia. Tumour vascularity was better estimated by measurement of lambda. Measurement of relative regional cerebral blood volume also correlated with tumor vascularity. Mainly the possibility of direct quantification of the Xe-CT data was advantagenous compared to perfusion MRI, and the XeCT data could be interpreted more clearly. Further upgrades of the technical equipment of MR as well as upgrades of available software will increase the applicability of perfusion MRI. However, both modalities offer a more precise investigation of tumor vascularization compared to ceMRI since potential dysplastic alteration may be detected earlier, which will be advantageous for planning bioptical approach of such tumours. PMID- 9861654 TI - [Appearance of the resection area of brain tumors in intraoperative MRI imaging]. AB - During MRI-controlled resection of brain tumors using an open MRI system, operation-induced alterations may occur, especially enhancement of the resection cavity wall. This may simulate tumor areas, resulting in false assessment of the resection or resection of non-tumorous areas. Based on 42 MRI- and biopsy controlled brain tumor resections in an 0.5 T open MRI (Signa SP, GE), the appearance and origin of operation-induced reactions are analyzed. In our opinion, there is a superposition of preformed peritumoral reactions by operation induced microcontusions. The beginning of the cavity wall enhancement needs at least 10-15 min. MRI-controlled analysis of the intraoperative steps by the neurosurgeon and neuroradiologist allows discrimination of operation-induced reactions from tumor areas and leads to safe operation. PMID- 9861655 TI - [Can superparamagnetic contrast media improve MRI-tomographic images of experimental gliomas?]. AB - PURPOSE: To investigate whether the margins of microscopic tumors can be delineated better with monocrystalline iron oxide nanoparticles (MION), a superparamagnetic contrast medium, than with Gd-DTPA by magnetic resonance imaging (MRI). METHODS: MRI and histological examinations were conducted in 28 Wistar rats with sterotactically implanted gliomas (C6 gliomas). Of the 28 animals, 14 were examined after intravenous administration of MION [nine animals received 179 mmol Fe/kg body weight (dose 1), and five, 893 mmol Fe/kg (dose 2)]. The other 14 animals were examined first after i.v. administration of Gd-DTPA (0.2 mmol/kg) and then after i.v. administration of MION. The extent of the tumors as seen on MRI and at histological study were compared. RESULTS: Iron particles were identified microscopically in tumor cells and in the tumoral interstitium. After administration of MION at dose 1, the contrast-enhanced area of tumor was 1.55-fold greater than the extent of tumor identified by histological study, at dose 2,2.15-fold. Compared with Gd-DTPA the area of contrast enhancement was greater by a factor of 1.38 with MION administration at dose 1 and by a factor of 1.91 at dose 2. CONCLUSION: MION provides intra- and extracellular contrast enhancement. The area of the contrast-enhanced tumor is dose-dependently greater with MION than with Gd-DTPA and also greater than the extent of tumor seen at histological study. PMID- 9861656 TI - [Effect of brain edema on the recurrence pattern of malignant gliomas]. AB - PURPOSE: To assess the influence of initial preoperative brain edema in malignant gliomas on regrowth patterns. SUBJECTS AND METHODS: 79 patients with histologically verified supratentorial malignant glioma were prospectively studied by magnetic resonance imaging (MRI) before and every 2-3 months after surgery. The median follow-up time was 11 months. We correlated the configuration of the initial vasogenic edema on T2-weighted images with tumor regrowth patterns on contrast-enhanced T1-weighted images. RESULTS: 35/47 tumor regrowths (75%) imitated the initial edema configuration, while 11/47 occurred within the initial tumor bed; in one case tumor recurrence was multilocal. CONCLUSION: In glioblastoma, tumor regrowth patterns correlate positively with the configuration of the initial vasogenic brain edema. The initial, "presurgical" peritumoral edema should thus be considered when planning further treatment. PMID- 9861657 TI - [Subperiostal hematoma of the orbit and epidural hematoma in patients with skull brain trauma]. AB - Subperiostal hematomas (SPOH) of the orbit are mostly of traumatic origin, but may occur spontaneously. In patients with frontal and temporal extradural hematomas, concomitant SPOH of the ipsilateral orbit have sporadically been observed. We report clinical and radiological findings of a patient and a review of the literature and discuss the development of SPOH. PMID- 9861658 TI - [CT-angiography as a non-invasive method for the evaluation of the patency of TIPSS (transjugular intrahepatic portasystemic shunt)]. AB - PURPOSE: To evaluate the feasibility of determining patency of the transjugular intrahepatic portosystemic shunt (TIPSS) by non-invasive CT angiography (CTA). MATERIALS AND METHODS: (1) Non-enhanced scanning of the shunt. (2) Bolus tracking by injecting 20 ml of non-ionic contrast material through a cubital vein access to determine the time to maximal shunt enhancement. (3) Contrast-enhanced spiral CT study applying a delay according to the time to peak of the shunt, 3 mm collimation, 5 mm table feed and 3 mm reconstruction interval. (4) 3D and multiplanar reconstructions. (5) Evaluation of the questions: intrahepatic shunt patent or not; evidence of intimal hyperplasia; evidence of stenosis and potential location. (6) Transjugular portography via the stent. (7) Comparison of angiographic findings and CT morphology. RESULTS: Eight patients had inconspicuous CTA. Four of them had a normal shunt at angiography, four had slight intimal hyperplasia. No intervention was necessary in these patients. CTA of three patients showed intimal hyperplasia (lumen reduction between 10% and 50%). The diagnosis was angiographically confirmed in all cases. Due to a high portosystemic gradient intervention was required in all. In five patients CTA and angiography showed a stenosis (reduction of shunt lumen > 50%). All required a revision including stent placement or PTA of the shunt tract. Four shunts were occluded; all occlusions were shown both in CTA and angiography. CONCLUSIONS: None of the shunts with normal findings at CTA required revision. All shunts conspicuous on CTA resulted in revision. In this study, CTA turned out to be an accurate, non-invasive method to evaluate the patency of TIPSS. PMID- 9861659 TI - [Neuroradiologic diagnosis of tumor in the inner nasal canthus]. PMID- 9861660 TI - [Imaging in intensive care. Methods, indications, diagnostic signs. I]. PMID- 9861663 TI - Carcinoma of the nasopharynx. AB - Nasopharyngeal carcinoma (NPC) is an intriguing disease that shows a distinct geographical distribution with well-defined, high-risk ethnic groups. This neoplasm exhibits strong associations with the Epstein-Barr virus, dietary and genetic factors. Radiation therapy is the mainstay of treatment and imaging plays a central role in tumor mapping and radiation therapy planning. NPC provides an excellent model for the study of normal and pathological anatomy of the skull base. It also provides an excellent model in the study of consequences of radiation therapy. PMID- 9861664 TI - Squamous cell carcinoma of the oropharynx and oral cavity: how imaging makes a difference. AB - The development of modern imaging techniques has significantly altered the treatment and management of these malignancies. Important treatment decisions that were once made intraoperatively are now made by using information from CT and MR imaging. The intent of this article is to provide the specific information that needs to be transmitted to the referring otolaryngologist or radiation oncologist and which will alter the treatment of patients with squamous cell carcinomas of the oral cavity and oropharynx. PMID- 9861665 TI - Imaging of squamous cell carcinoma of the hypopharynx. AB - The hypopharynx is a clinically silent area and early lesions may be asymptomatic for a long period. At presentation, primary squamous cell carcinoma of this area is usually advanced. Almost all of these lesions are studied with imaging as part of the clinical work-up. The goal of this article is to help the practicing radiologist convey a report which provides information that will directly influence treatment of patients with hypopharyngeal carcinoma. PMID- 9861666 TI - Imaging of laryngeal cancer. AB - CT and MRI are the main modalities for examination of laryngeal pathology. Generally, MRI seems to be the optimal method of examination in cooperative patients, especially for evaluation of their larynx before an attempted partial laryngectomy. The choice between the two modalities will also be determined by one's experience with these modalities. The possibilities of CT and MRI vary clearly from each other regarding detection of cartilage invasion. MRI seems to be more sensitive than CT in detection of neoplastic cartilage invasion, but seems to have a somewhat lower specificity, especially for thyroid cartilage involvement. There are increasing indications for imaging regarding tumor volume, and signs of cartilage involvement may have prognostic significance for the risk of tumor recurrence. PMID- 9861667 TI - Nuclear migration, nucleokinesis and lissencephaly. AB - During the past 20 years, biologists have become used to finding that proteins first identified in simple, genetically manipulable eukaryotic organisms are conserved in higher eukaryotes. This article draws attention to the similarity between NUDF protein, which is required for nuclear migration in the filamentous fungus Aspergillus nidulans, and a mammalian homologue, LIS1, whose malfunction causes lissencephaly, a neuronal migration disease. The authors suggest that there might be an underlying similarity of mechanism between nuclear migration in the fungus and neuronal migration in the brain. PMID- 9861668 TI - NSF--fusion and beyond. AB - NSF (N-ethylmaleimide-sensitive fusion protein) was the first protein to be isolated as a crucial factor in intracellular membrane-fusion events, such as the fusion of synaptic vesicles with the presynaptic membrane during neurotransmission. Although the activation of membrane SNARE proteins for subsequent fusion is clearly a primary role of NSF, recent studies have provided surprising evidence that NSF also interacts with glutamate receptors at the postsynaptic membrane in a way that does not seem to involve SNAREs. These results suggest that NSF might act as a molecular chaperone not only on SNAREs but also of other proteins. PMID- 9861669 TI - Diverse functions of vertebrate gap junctions. AB - Gap junctions are clusters of intercellular channels between adjacent cells. The channels are formed by the direct apposition of oligomeric transmembrane proteins, permitting the direct exchange of ions and small molecules (< 1 kDa) between cells without involvement of the extracellular space. Vertebrate gap junction channels are composed of oligomers of connexins, an enlarging family of proteins consisting of perhaps > 20 members. This article reviews recent advances in understanding the structure of intercellular channels and describes the diverse functions attributable to gap junctions as a result of insights gained from targeted gene disruptions in mice and genetic disease in humans. PMID- 9861670 TI - Molecular mechanisms of DNA double strand break repair. AB - DNA double-strand breaks (DSBs) are major threats to the genomic integrity of cells. If not taken care of properly, they can cause chromosome fragmentation, loss and translocation, possibly resulting in carcinogenesis. Upon DSB formation, cell-cycle checkpoints are triggered and multiple DSB repair pathways can be activated. Recent research on the Nijmegen breakage syndrome, which predisposes patients to cancer, suggests a direct link between activation of cell-cycle checkpoints and DSB repair. Furthermore, the biochemical activities of proteins involved in the two major DSB repair pathways, homologous recombination and DNA end-joining, are now beginning to emerge. This review discusses these new findings and their implications for the mechanisms of DSB repair. PMID- 9861671 TI - Make room for dynein. AB - Three classes of cytoskeletal motor protein have been identified--myosins, kinesins and dyneins. Together, these proteins are now thought to be responsible for the remarkable variety of movements that occur in eukaryotic cells and that are essential for reproduction and survival. Crystallographic analysis of the myosin and kinesin motor domains at atomic resolution has provided insight into their mechanism of force production. However, because of its relative intractability to molecular manipulation, definition of the dynein motor domain, let alone progress in understanding how it works, has been slower. Evidence now indicates that the microtubule-binding domain of dynein is spatially isolated from the ATPase domain at the tip of a projecting coiled coil. As proposed here, this curious arrangement might serve to accommodate multiple copies of the outsized and functionally complex motor heads on the microtubule surface. PMID- 9861672 TI - Old and new pathways of protein export in chloroplasts and bacteria. AB - Targeting of chloroplast proteins to the thylakoid membrane is analogous to bacterial secretion, and much of what we know has been learned from secretory mechanisms in Escherichia coli. However, chloroplasts also use a delta pH dependent pathway to target thylakoid proteins, at least some of which are folded before transport. Previously, this pathway seemed to have no cognate in bacteria, but recent results have shown that the HCF106 gene in maize encodes a component of this pathway and has bacterial homologues. This delta pH-dependent pathway might be an ancient conserved mechanism for protein translocation that evolved before the endosymbiotic origin of plastids and mitochondria. PMID- 9861673 TI - Pictures in cell biology. Structures of fusion-machinery components. PMID- 9861674 TI - Analysis of the endocytic system by using horseradish peroxidase. PMID- 9861675 TI - The 1997 list. Proposed new bacterial taxa and proposed changes of bacterial names published during 1997 and considered to be of interest to medical or veterinary bacteriology. An informational note. AB - A list of names published or validated in 1997 is presented. We also comment on the tendency to base names of new taxa on a single bacterial strain, and the consequences for reliable descriptions that this tendency implies. PMID- 9861676 TI - Catalase-positive Eikenella corrodens and Eikenella-like isolates of human and canine origin. AB - Ten catalase-positive isolates and one catalase-negative isolate that had been assigned to Eikenella corrodens were compared to the nomenclatural type strain regarding selected phenotypic and molecular features and chromosomal deoxyribonucleic acid (DNA) relatedness using the spectrophotometric method. Five catalase-positive human isolates were assigned to the genomic species Eikenella corrodens on the basis of high DNA relatedness levels. Three others, among them strain Chen UB 204, exhibited only moderate degrees of DNA relatedness to the type strain and with each other. Two catalase-positive isolates from dogs were closely interrelated, but yielded only low degrees of DNA binding with Eikenella corrodens and the Eikenella-like human isolates. These findings confirm that the human eikenellas comprise more than one genomic species and that the canine strains represent a distinct taxonomic entity. The differentiation of the strains investigated by conventional phenotypic features, hydrolytic enzyme reactions, and cellular carbohydrate patterns was considered. PMID- 9861677 TI - Regrowth of Legionella pneumophila in a heat-disinfected plumbing system. AB - We examined the factors involved in the occurrence of Legionellaceae in a hospital water system and the recontamination by Legionella pneumophila after a thermal disinfection procedure was studied. Three months after the heat treatment (70 degrees C), the regrowth of the two prevalent Legionella strains (L. pneumophila serogroup 1 [Oxford-like] and L. pneumophila serogroup 2) reached the original level of cell numbers. Genomic analysis (pulsed-field gel electrophoresis) revealed the strains to be survivors of the decontamination. Temperature tolerance experiments showed that the serogroup 1 strain exhibited a higher tolerance to 60 degrees C than the serogroup 2 strain, which could account for the order of reappearance of the strains after the heat treatment. Potential host amoebae, including Acanthamoeba spp. and Vahlkampfia spp., which are known to play a critical role in the amplification process of Legionella, were isolated from the plumbing system. In-vitro studies demonstrated both Legionella strains for a similar rate of multiplication in A. castellanii. In competitive coinfections, however, the serogroup 1 strain achieved a higher rate of multiplication if compared with the serogroup 2 strain. PMID- 9861678 TI - Selective isolation of Pseudomonas stutzeri from vertebrate faeces on Rambach agar. AB - Faecal samples collected from 308 wild birds of 25 species and 19 rodents of 3 species in South Moravia (Czechland) were pre-incubated in Muller-Kauffmann tetrathionate broth at 42 degrees C for 24 h and then streaked onto Rambach agar plates which were incubated at 37 degrees C for 48 h. Seventeen out of 22 isolates forming orange-red colonies on Rambach agar were identified as Pseudomonas stutzeri, the rest as Pseudomonas sp. and Alcaligenes sp. The colonies of P. stutzeri were either dry, wrinkled and adherent to the agar (resembling Bacillus) or smooth, less adherent (mimicking Salmonella). P. stutzeri was recovered from five species of vertebrates caught in farmland habitats: the house sparrow (Passer domesticus), the tree sparrow (P. montanus), the great warbler (Acrocephalus arundinaceus), the wood mouse (Apodemus sylvaticus) and the common vole (Microtus arvalis). The overall isolation rate was 4.5% in birds (12.6% in house sparrows) and 15.8% in rodents. The procedure can be useful for the isolation of P. stutzeri in clinical and environmental studies. PMID- 9861679 TI - Diversity among clinical isolates of Proteus penneri detected by random amplified polymorphic DNA analysis. AB - DNA of thirteen haemolytic Proteus penneri strains of clinical origin, all producing calcium dependent haemolysin and having been derived from four European countries was examined for plasmid profile, and outer membrane protein profile, by random amplified polymorphic DNA-PCR (RAPD-PCR) method, and digestions with restriction endonucleases were performed. All strains contained two large plasmids of approximately 60 and 70 kilobase pairs (kb). In addition, four strains contained a small plasmid of about 6 kb. These four strains produced cell bound haemolysin only. Outer membrane protein analysis revealed subtle differences between strains. RAPD-PCR with primer I (CCGCAGCCAA) revealed 13 types, whereas primer II (AACGCGCAAC) yielded only two main types of different patterns. Results with primer I suggests a DNA sequence diversity within this species. The RAPD-PCR method provides a fast, economical and reproducible means for the typing of P. penneri. Digestion with restriction endonucleases indicated a high level of DNA methylation in this species. PMID- 9861680 TI - Fibronectin and laminin binding of urogenital and oral prevotella species. AB - 88 strains of five Prevotella species--P. bivia, P. buccae, P. disiens, P. oralis, and P. oris--were examined for their fibronectin and laminin binding properties with the aid of latex particle agglutination assays. Beside single protein binding activities, all species showed strains that adhered to both fibronectin and laminin. The oral species, P. buccae, P. oralis, and P. oris were found to interact with laminin to a pronouncedly higher extent than with fibronectin. The urogenital species, P. bivia and P. disiens showed comparable activities of binding to fibronectin and laminin, with P. bivia exhibiting higher matrix protein binding rates than P. disiens. Within the oral species group, P. oralis showed a higher percentage of fibronectin and laminin reactive strains than did P. buccae and P. oris. The finding of species-related different binding properties may throw some light on the known differences in clinical relevance and pathogenicity of the urogenital species, P. bivia and P. disiens, but does so only in part concerning the oral species, P. buccae, P. oralis, and P. oris. Moreover, the observed differences in matrix protein binding of Prevotella species may have implications in chemotaxis and opsonization on the one hand and maintenance of colonization activities under antibiotic therapy on the other. PMID- 9861681 TI - Coagulase-negative variants of methicillin-resistant Staphylococcus aureus subsp. aureus strains isolated from hospital specimens. AB - At the Department of Clinical Bacteriology of Medical University of Warsaw, S. aureus strains displaying negative reactions in the coagulase tube-test were isolated with a relatively high frequency from clinical specimens. As many as seventeen of such strains all from different patients were isolated during 1994 among MRSA strains. Despite coagulase negativity all strains were positive in tests for clumping factor (CF), protein A and thermonuclease. Biochemical characteristics, phage patterns and antibiotic resistance characteristics of these strains were tested. Most of the coagulase-negative strains possessed a heterogenous type of methicillin resistance. Apart from methicillin most of them were resistant to many other antimicrobials. All were resistant to gentamicin, tetracyclines and macrolide-lincosamide-streptogramine B. An occurrence of coagulase-negative S. aureus strains may lead to problems in their identification and the necessity of an application of other methods like CF, protein A or biochemical reactions. PMID- 9861682 TI - Effect of purified staphylococcal leukocidal toxins on isolated blood polymorphonuclear leukocytes and peritoneal macrophages in vitro. AB - Bicomponent (fractions S and F) staphylococcal leukocidal toxins (Panton Valentine leukocidin-Luk and haemolysin gamma-Hlg) were tested for in vitro activity against isolated polymorphonuclear leukocytes (PMNL) and peritoneal macrophages (PMF). For assessment of membrane permeability at subcytolytic concentrations of leukocidin (Luk-S + Luk-F) and haemolysin gamma (HlgA + HlgB) (8-1000 ng/ml), PMNL and PMF were radiolabelled (86Rb, 14C-amino-isobutyric acid (AIB) or 51Cr). All toxins tested caused lysis of human PMNL, although considerable differences were noted in the sensitivity of these cells to Luk and Hlg. Release of 51Cr (at 1000-5000 ng/ml), being a sign of irreversible cell damage and lysis, was preceded, at lower concentrations of the toxins (40 and 200 ng/ml), by the release of large amounts of low-molecular labels--86Rb and 14C AIB. In another experiment, it was found that release of 86Rb from PMNL incubated with low concentrations of Luk (50 ng/ml) took place after 15-30 minutes of incubation, when no significant amounts of 14C-AIB or 51Cr were released. These findings support the concept of pore formation by staphylococcal leukocidal toxins in membranes of sensitive cells and indicate that a relatively short time is needed for the formation of these pores after binding of the Luk-S and Luk-F components to the membrane. PMID- 9861683 TI - Inhibition of bacterial pathogens by lactobacilli. AB - Lactobacilli produce many antimicrobial substances including organic acids, hydrogen peroxide and bacteriocins. Antagonistic activity of lactobacilli is an important factor in the protection of the vagina of fertile women against infection by certain pathogens. In the present study, we investigated 17 strains of lactobacilli, including 11 strains of vaginal origin. The aim was to investigate in more detail the antibacterial activity of lactobacilli and to attempt to assess substances responsible for inhibition. The investigated lactobacilli inhibited some strains of Escherichia coli, Serratia marcescens, Shigella boydii, Listeria monocytogenes, Listeria ivanovii, Listeria innocua and Staphylococcus aureus. We have provided evidence that inhibition is due mainly to organic acids and to a lesser extent, to bacteriocins. PMID- 9861685 TI - Antibodies to the E2 protein of GB virus C/hepatitis G virus: low prevalence in Asian countries. AB - Epidemiological investigations of GB virus C (GBV-C)/hepatitis G virus (HGV), an infectious agent discovered in 1995/1996, are facilitated by a recently developed immunoassay for the detection of antibodies to the viral envelope 2 protein (anti E2). We used this assay to establish GBV-C/HGV prevalence in seven European, African, and Asian countries. A total of 1579 serum samples from healthy adults lacking prior exposure to known parenteral risk factors was screened. Anti-E2 positivity ranged from 13.6% (Italy) to 7.7% (Mauritius) in the European and African countries investigated. Anti-E2 prevalence was exceedingly low in the Philippines and Sri Lanka. This observation might be attributable to socio economical and demographic factors. PMID- 9861686 TI - Rate of carriage, serotype distribution and penicillin resistance of Streptococcus pneumoniae in healthy children. AB - This study was aimed to define the carriage rates for Streptococcus pneumoniae in a given population in Ankara and also to determine the serotypes and penicillin resistance of these strains. Oropharyngeal swabs were taken from a total of 661 children aged between 0-11 years and living in a province of Ankara between January 1995-January 1997. Serotyping was performed by detection of the Quellung reaction. The isolates were screened for penicillin susceptibility by the agar dilution method according to the guidelines of NCCLS. The total rate of pneumococcal carriage in the study population was 23.90% and the isolation rate was found to be statistically associated with age, being higher in small children. Among the 158 S. pneumoniae isolates, the most prevalent serotypes (in order of frequency) were 6, 19, 9, 23, 3 and 14. Penicillin susceptibility was examined in 120 of the isolates. 55 of them (45.83%) were susceptible, 53 (44.17%) were intermediately and 12 (10.0%) were highly resistant to penicillin. Evaluation of the results showed that serotypes 6, 14 and 23 were those most often associated with penicillin resistance. The significant rate of isolation of penicillin-resistant pneumococci in healthy carriers points to the importance of active immunization in risk groups and also the importance of the rational use of antibiotics to limit the spread of resistant strains. PMID- 9861687 TI - FT infrared and Raman investigation of saccharide-phosphatidylcholine interactions using novel structure probes. AB - Conformational consequences of adduct formation between saccharides (trehalose, glucose, raffinose) and sorbitol with dipalmitoylphosphatidylcholine (DPPC) in multibilayers are revealed by relative intensity changes of the band components corresponding to the nu asN(CH3)3 and nu sC-N(CH3)3 stretching modes of the choline chain terminal and those of the nu C = O band. The conformational sensitivity of those modes was demonstrated previously (J. Grdadolnik et al., Chem. Phys. Lipids 65 (1993) 121) and used to demonstrate the effects of stepwise hydration of phosphatidylcholines. The latter are compared with the effects of saccharide binding and found to be qualitatively similar, but not identical. The same is true of the low frequency shifts of the nu asPO2- vibration: the shifts due to saccharide binding correspond to the binding of six to seven water molecules per phosphate which is about 20 cm-1 less than the shift caused by full hydration. A particularly interesting finding concerns the appearance of two bands in the nu asPO2- region of the DPPC-saccharide adducts. The relative intensities of the two bands (1243 and 1223 cm-1) change on additional hydration; it is the one at 1223 cm-1 that prevails at high hydration levels. Major changes in saccharide conformation are not detectable but minor differences between the DPPC bound and crystal spectra are observed. PMID- 9861684 TI - Cytokine synthesis of human monocytes stimulated by triple or single helical conformer of an antitumour (1-->3)-beta-D-glucan preparation, sonifilan. AB - It has been suggested that the immunopharmacological activity of soluble (1-->3) beta-D-glucan depends on it's conformation in mice. In this study, we examined the relationship between the conformation of Schizophyllan (SPG), a high molecular weight (1-->3)-beta-D-glucan, and cytokine productivity in an in vitro human system. Monocyte-like human cell lines, THP-1 and U-937, and peripheral blood mononuclear cells (PBMC) were used. THP-1 and U-937 cells were differentiated by phorbol myristate acetate (PMA) before use. SPG usually has a triple helical conformation in water, but it was modified by treatment with aqueous sodium hydroxide to become a single helical conformer (SPG-OH). SPG or SPG-OH was added to the macrophage cell culture and gene expression and translation of several cytokines was analyzed by RT-PCR, ELISA, or bioassays. Differentiated THP-1 expressed high levels of cytokine genes, such as IL-8, in response to SPG-OH. High levels of IL-12 p70 were detected from THP-1 cells stimulated with SPG-OH. U-937 cells expressed high levels of IL-8 and TNF-alpha after SPG-OH treatment. Furthermore, PBMC isolated from healthy donors also strongly reacted with SPG-OH but not with SPG. High concentrations of TNF-alpha were detected in SPG-OH-stimulated PBMC cultures. These data suggest that the biological activities of SPG are strongly associated with its conformation in humans. PMID- 9861688 TI - A spectrophotometric method for the determination of hydroperoxides in liposomes. AB - A modification of the Asakawa-Matsushita iodometric assay method for the determination of the content of lipid hydroperoxides was developed which permits the simultaneous processing of many samples of high lipid content. The method has the advantages of simplicity as well as good reproducibility, so it is not necessary to process standards with each determination. Our technique exceeds the sensitivity attained with other spectrophotometric determinations reported in the literature. The method requires the total elimination of water from the samples, and this was accomplished using an azeotropic mixture of ethanol:water of 96:4. The results obtained with liposomes indicate that the method is applicable to biological material limited to small volume samples, ranging 5-50 microliters. We want to emphasize that this method permits the study of the peroxidation process as function of time. PMID- 9861689 TI - Study on a fluorescence-enhanced system of 1,2,4-trihydroxyanthroquinone-Be(2+) DNA. AB - Fluorescence-enhancement of 1,2,4-trihydroxyanthroquinone (THAQ)-Be2+ complex enhanced by nucleic acid was studied. Experimental results revealed that double stranded DNA can enhance remarkably the fluorescence intensity of THAQ-Be2+, while RNA cannot. Based on these results, a fluorescence method for the selective determination of DNA in the presence of RNA was developed. Maximum fluorescence intensity was found in the pH range 3.6-4.5 with maximum excitation and emission wavelengths at 510 and 565 nm, respectively. Under optimum conditions, the calibration graph was linear over the range 0.08-18 micrograms ml-1 for double stranded fish sperm DNA (fsDNA) with the detection limit being 2.75 x 10(-8) g ml 1. The method was applied for the determination of DNA in synthetic samples. The relative S.D. for five replicates was within 4%. In addition, the interaction mechanism of THAQ-Be2+ with DNA was also discussed. PMID- 9861690 TI - Characteristics of calls to Israeli hotlines during the Gulf War. AB - The current study presents results of a survey of 3,215 calls received at seven centers of telephone emergency services (TES) in Israel during the Gulf War, when citizens of Israel experienced severe stress resulting from SCUD missile attacks. Whereas former surveys have shown that characteristics of calls to TES in Israel are generally not affected by external stressogenic events, a remarkable change was recorded in both the quantity and quality of calls received in TES centers in Israel during the Gulf War. The relative frequencies of problem categories presented by callers during the Gulf War revealed a significant increase in "environmental pressures," a category that reflected the stressful situation of the war, as opposed to intra- or interpersonal problems typical of peacetime calls. A comparison between this group of "war calls" and a control group of "nonwar calls," revealed that the two groups represented populations of callers differing in sociodemographic characteristics, expectations, and benefits from the calls. Results are discussed in reference to the unique role of TES as a source of psychological first-aid in a community crisis situation. PMID- 9861691 TI - Culturally sensitive AIDS educational videos for African American audiences: effects of source, message, receiver, and context. AB - The importance of using culturally sensitive educational materials in HIV-related interventions with racial and ethnic minority groups is widely recognized. However, little empirical research has been conducted to assess the relative effectiveness of different techniques for creating culturally sensitive AIDS educational videos. Two field experiments with three samples of African American adults (N = 174, 173, and 143) were conducted to assess how source characteristics (race of communicator), message characteristics (multicultural message vs. culturally specific message), and audience characteristics (racial distrust and AIDS-related distrust) influence proximate (perceptions of the message's credibility and attractiveness) and distal (AIDS-related attitudes, beliefs, and behavioral intentions) output variables for AIDS educational videos. In Study 1, an AIDS video with a culturally specific message was rated as more credible, more attractive, and of higher quality than was a video with a multicultural message. The multicultural message was rated less favorably when delivered by a White announcer than when the announcer was Black. In Study 2, the same pattern was replicated with a second community sample and a campus-based sample. Study 2 also indicated that a multicultural message might be more effective if delivered in a culturally specific context, namely, after audience members watch a culturally specific video. Minimal changes were observed in distal outcome variables. It is argued that influencing proximate output variables is necessary, though not sufficient, for effecting long-term change in AIDS-related attitudes, beliefs, and behaviors. PMID- 9861692 TI - Out on campus: sexual orientation and academic climate in a university context. AB - Although lesbian, gay, and bisexual (LGB) students often "come out" in university settings, empirical studies have demonstrated that these environments are often hostile toward them. The current paper posits that such hostile contexts adversely affect their educational experiences. Results from a survey of a stratified random sample of 1,927 undergraduate and graduate students on a scale measuring perceptions of academic climate (General Campus Climate) supported this claim: LGB students had more negative perceptions of a variety of campus factors. In addition, a scale measuring perceptions of Lesbian, Gay, and Bisexual Campus Climate found that LGB students were more likely than heterosexuals to perceive the campus as inhospitable to LGB people. Women and racial and ethnic minorities were also more likely to rate the LGB climate as hostile. Finally, data from two scales measuring attitudes believed to influence these perceptions yielded gender, racial/ethnic, religious, and sexual orientation differences in theoretically meaningful directions. Results indicate that LGB students often experience the university in more negative ways than heterosexual students, and that certain campus communities--particularly women and racial/ethnic minorities- are more aware of this negativity and contribute to it less because of their relatively more progay attitudes. Implications for how to improve LGB students' experiences are discussed. PMID- 9861693 TI - Evaluation of indicated preventive intervention (secondary prevention) mental health programs for children and adolescents. AB - Evaluated the outcomes of 130 indicated preventive interventions (secondary prevention) mental health programs for children and adolescents that seek to identify early signs of maladjustment and to intervene before full-blown disorders develop. Results indicate such programs significantly reduce problems and significantly increase competencies. In particular, behavioral and cognitive behavior programs for children with subclinical disorders (mean ESs in the 0.50s) appear as effective as psychotherapy for children with established problems and more effective than attempts to prevent adolescent smoking alcohol use, and delinquency. In practical terms, the average participant receiving behavioral or cognitive-behavior intervention surpasses the performance of approximately 70% of those in a control group. Of particular interest was the high mean effect (0.72) achieved by programs targeting incipient externalizing problems which are customarily the least amenable to change via traditional psychotherapeutic efforts when they reach clinical levels. Priorities for future research include greater specification of intervention procedures, assessment of treatment implementation, more follow-up studies, and identifying how different participants respond to early intervention. PMID- 9861694 TI - Color and lightness constancy in different perceptual tasks. AB - Color and lightness constancy with respect to changing illumination was studied with three different perceptual tasks: ranking of colored papers according (1) to their lightness and (2) to their chromatic similarity in photopic, mesopic, and scotopic states of adaptation, and (3) recognition of remembered colored papers after changes of illumination in photopic vision. Constancy was found in the second task, only. Excitations of light receptors and luminance channels were computed to simulate the empirical rank orders. Results of the first task can be predicted with the hypothesis that luminance channels are activated, if lightness is asked for. Sequences arranged with respect to chromatic similarity were found independent of the illuminant spectra, even if the calculated rank orders of cone excitation were changed in the altered illumination. PMID- 9861696 TI - Chemoenzymatic synthesis of an N-acetylneuraminic acid analogue having a carbamoylmethyl group at C-4 as an inhibitor of sialidase from influenza virus. AB - 5,9-Diacetamido-2,6-anhydro-O-4-carbamoylmethyl-3,5,9-trideo xy-D-glycero- D galacto-non-2-enonic acid (1) was synthesized via a key intemediate 2 through the Neu5Ac aldolase [E.C.4.1.3.3]-catalyzed aldol reaction of 2-acetamido-2,6-dideoxy 6-azido-D-glucose with sodium pyruvate operating under alkaline conditions (pH 10.5) in order to accelerate epimerization C-2 of N-acetyl-D-glucosamine (D GlcNAc) derivatives. Compound 1 showed inhibitory activity against sialidase. PMID- 9861697 TI - Analysis of specific interactions of synthetic glycopolypeptides carrying N acetyllactosamine and related compounds with lectins. AB - Analysis of interactions of synthetic glycopolypeptides with lectins was performed with a biosensor based on surface plasmon resonance (SPR). A series of synthetic oligosaccharide-substituted poly(L-glutamic acid)s were immobilized on sensor surfaces via the gamma-carboxyl groups of their peptide moieties by the surface thiol coupling method. Artificial glycopolypeptides: an N acetyllactosamine-substituted polymer (1), an N-acetylisolactosamine-substituted polymer (2), a (GlcNAc)3-substituted polymer (3), a (GlcNAc)2-substituted polymer (4), and a p-aminophenyl N-acetyl-beta-lactosaminide-substituted polymer (5), were used as the ligands. On analysis by SPR, surface-bound polymers 1 and 5 reacted with Erythrina cristagalli agglutinin (ECA), Lycopersicon esculentum agglutinin (LEA), Ricinus communis agglutinin-120 (RCA120), and wheat germ (Triticum vulgaris) agglutinin (WGA). Polymer 2 reacted with WGA and RCA120, but did not with ECA and LEA. The results indicate that beta-(1-->4)-linked galactosyl residues are needed for binding to ECA and LEA. Polymer 3 reacted strongly with LEA and WGA, but polymer 4 reacted strongly only with WGA. Affinity constants (KA) for surface-bound polymer 5-lectin interactions were also about 4 61 times as strong as those for surface-bound polymer 1-lectin interactions. These artificial glycopolypeptides were shown to be useful as tools and probes of carbohydrate recognition and modeling in the analysis of glycoprotein-lectin interactions. PMID- 9861698 TI - Studies towards neoglycoconjugates from the monosaccharide determinant of Vibrio cholerae O:1, serotype Ogawa using the diethyl squarate reagent. AB - The effect of reaction time, concentration and molar excess of hapten upon the efficiency of the conjugation of carbohydrates to proteins using the diethyl squarate reagent has been studied using chicken serum albumin (CSA) as the carrier protein and a linker-equipped D-glucose derivative as the hapten. A high degree of incorporation of the latter into CSA was achieved with high efficiency, and the use of a large excess of the ligand was not necessary. Conjugation of the immunodominant monosaccharide determinant of Vibrio cholerae O:1, serotype Ogawa, bearing the same spacer, followed a similar pattern, showing that the nature of the carbohydrate does not substantially affect the outcome of the conjugation and that a predicted degree of antigen-loading onto carrier protein is possible to achieve. PMID- 9861699 TI - Isolation and characterisation of disodium (4-amino-4-deoxy-beta-L- arabinopyranosyl)-(1-->8)-(D-glycero-alpha-D-talo-oct-2-ulopyranosylona te)- (2- >4)-(methyl 3-deoxy-D-manno-oct-2-ulopyranosid)onate from the lipopolysaccharide of Burkholderia cepacia. AB - A trisaccharide was isolated from the core oligosaccharide in the lipopolysaccharide (LPS) of Burkholderia cepacia GIFU 645 (ATCC 25416, type strain) by methanolysis followed by HPLC and saponification. It was identified by MS, methylation analysis and 1H and 13C NMR spectroscopy as disodium (4-amino-4 deoxy-beta-L-arabinopyranosyl)-(1-->8)-(D-glycero- alpha-D-talo-oct-2 ulopyranosylonate)-(2-->4)-(methyl 3-deoxy-D-manno-oct-2-ulopyranosid)onate. In addition to the trisaccharide derivative, methanolysis gave dimethyl (D-glycero alpha-D- talo-oct-2-ulopyranosylonate)-(2-->4)-(methyl 3-deoxy-D-manno-oct-2- ulopyranosid)onate in a relative proportion to the trisaccharide of 3:1, indicating a non-stoichiometric (approximately 25%) substitution of the octulosonic acid by 4-amino-4-deoxyarabinose in the LPS. PMID- 9861700 TI - Antibody recognition of epitopes on wild-type and mutant beta-(1-->4) galactosyltransferase-1. AB - The epitopes present on beta-(1-->4)-galactosyltransferase-1 (beta 4Gal-T1) have been explored using a panel of monoclonal antibodies (mAbs) raised against the soluble form of the human enzyme. Reactivity of the antibodies with site-specific and truncated mutants of human beta 4Gal-T1 suggests the presence of a major immunogenic epitope cluster consisting of four epitopes within the stem region and mapping between amino acids 42 and 115. The catalytic activity of the enzyme is increased in the presence of stem region-specific antibody. Two of the epitopes were further localized to a region between amino acids 42 and 77, sequences which are not shared with the recently cloned beta 4Gal-T2 and beta 4Gal-T3 enzymes. An epitope located close to or within the catalytic domain is also identified, and the mAb to this region binds synergistically with antibodies to the stem region. PMID- 9861701 TI - Structure of the O-specific polysaccharide from Pseudoalteromonas elyakovii sp. nov. CMM 162. AB - The structure of the O-specific polysaccharide from lipopolysaccharide of Pseudoalteromonas elyakovii sp. nov. CMM 162 on the basis of NMR data, Smith degradation and methylation study was elucidated as follows:-->2)-alpha-D-Glcp-(1 ->4)-beta-D-GalpNAc-(1--> 3)-alpha-D-Galp-(1-->3)-beta-D-Galp-NAc-(1-->6)-alpha-D Glcp-(1-->. PMID- 9861702 TI - Vibrational circular dichroism: an incisive tool for stereochemical applications. AB - Vibrational circular dichroism (VCD) is the extension of circular dichroism (CD) from electronic to vibrational transitions in molecules. In this review, we present the basic concepts of VCD and focus on its growing strength for stereochemical applications. There are three principal areas of application of VCD. These are the determination of optical purity, the determination of absolute configuration and the determination of conformational properties of chiral molecules in solution. Until recently, the application of VCD was restricted to the use of instruments that had been custom assembled in academic research laboratories. This situation has now changed. It is possible to purchase a dedicated Fourier transform infrared (FTIR) VCD spectrometer complete with user friendly software. Combined with rapid advances in the ab initio theoretical prediction of infrared (IR) and VCD spectra, the availability of such technology opens the way for the widespread use of VCD spectroscopy for research and industrial applications. PMID- 9861703 TI - Use of CD and FT-IR to determine the secondary structure of purified proteins in the low-microgram range. AB - The spectroscopic characterization of protein secondary structure is often partially unreliable when samples are not extremely pure and abundant. This problem may be overcome by the combination of circular dichroism (CD) and Fourier transform infrared spectroscopy (FT-IR). We used these methods to characterize the secondary structure of two proteins of neurobiological interest, calexcitin (CE) and the cellular isoform of prion protein (PrPC). Both proteins were purified with multiple chromatographic steps and were obtained in buffer with high purity (> 95%) and in low amount (approximately 2 micrograms). The samples were analyzed by circular dichroism (down to 184 or 182 nm), recovered, and deposited on films for infrared analysis. The spectral deconvolution from the two methods yielded secondary structures in good agreement with each other as well as with theoretical predictions based on amino acid sequence. The conformation of CE was found to be dependent on its concentration and on calcium binding. The secondary structure of cellular native PrP varied dramatically with the detergent used. In conclusion, the combination of CD and FT-IR analysis is suitable for the characterization of the conformational changes induced by ligand binding and/or by different solvent conditions when the protein of interest is only scarcely available. The methods used here provide valuable insights into the putative correlation between protein structure and activity. PMID- 9861704 TI - Corticofugal regulation of auditory sensitivity in the bat inferior colliculus. AB - Under free-field stimulation conditions, corticofugal regulation of auditory sensitivity of neurons in the central nucleus of the inferior colliculus of the big brown bat, Eptesicus fuscus, was studied by blocking activities of auditory cortical neurons with Lidocaine or by electrical stimulation in auditory cortical neuron recording sites. The corticocollicular pathway regulated the number of impulses, the auditory spatial response areas and the frequency-tuning curves of inferior colliculus neurons through facilitation or inhibition. Corticofugal regulation was most effective at low sound intensity and was dependent upon the time interval between acoustic and electrical stimuli. At optimal inter-stimulus intervals, inferior colliculus neurons had the smallest number of impulses and the longest response latency during corticofugal inhibition. The opposite effects were observed during corticofugal facilitation. Corticofugal inhibitory latency was longer than corticofugal facilitatory latency. Iontophoretic application of gamma-aminobutyric acid and bicuculline to inferior colliculus recording sites produced effects similar to what were observed during corticofugal inhibition and facilitation. We suggest that corticofugal regulation of central auditory sensitivity can provide an animal with a mechanism to regulate acoustic signal processing in the ascending auditory pathway. PMID- 9861705 TI - Inhibitory receptor binding events among the components of complex mixtures contribute to mixture suppression in responses of olfactory receptor neurons of spiny lobsters. AB - Responses of olfactory receptor neurons of spiny lobsters Panulirus argus to two component mixtures can be shaped by inhibitory events such as odor-activated hyperpolarizations and inhibition of odor-receptor binding (Daniel et al. 1996). In the current study, we extend this analysis to complex mixtures by examining responses of spiny lobster olfactory receptor neurons to mixtures containing up to seven odorants, consisting of adenosine-5'-monophosphate, ammonium, betaine, L cysteine, L-glutamate, DL-succinate, and taurine. The response to a mixture was often less than the response to its most excitatory component. The effect of adding an excitatory odorant to a mixture depended on olfactory receptor neuron type, composition of the mixture, and which compound was added. In some cases the added excitatory compound had no effect or even decreased the mixture's response intensity, thus demonstrating nonlinear contributions of the components. Response intensities predicted by a noncompetitive model, which is most representative of these olfactory receptor neurons, were improved when the model included a term for empirical measurements of inhibitory binding interactions, suggesting that inhibitory binding interactions are one mechanism contributing to mixture suppression. This model's predictions were accurate for binary mixtures but not for larger mixtures, suggesting that additional inhibitory mechanisms are needed to account for mixture interactions in complex mixtures. PMID- 9861706 TI - Neurotransmitters alter the numbers of synapses and organelles in photoreceptor terminals in the lamina of the housefly, Musca domestica. AB - Various organelles in the lamina terminals of housefly photoreceptors exhibit daily rhythms having a circadian basis. These include changes in the numbers of photoreceptor tetrad and L2 feedback synapses, and longitudinal movements of screening pigment. Circadian information has previously been suggested to spread from the clock to the lamina via widefield cells expressing either 5 hydroxytryptamine or pigment-dispersing hormone-like immunoreactivity. We examined the action of these neuromodulators, and other candidate neurotransmitters, 4 h after injecting either the transmitter or a control into the medulla. We counted electron microscope profiles of organelles that normally exhibit circadian changes, and two types of invagination into photoreceptor terminals, capitate projections and inter-receptor invaginations. No single substance mediated the changes observed. Injected pigment-dispersing hormone peptide decreased the number of pigment granules, implicating this peptide in screening pigment migration, but produced no changes in synapse-related organelles. alpha-Aminobutyric acid exclusively decreased the number of L2 feedback synapses. Responses to other transmitters were specific, and often large, but generally not statistically significant. Histamine, for example, may decrease the number of tetrads, possibly by direct autoregulation. The results suggest that there is likely to be more than one effector in the circadian pathways to the lamina. PMID- 9861707 TI - Assessing the roles of glutamatergic and cholinergic synaptic drive in the control of fictive swimming frequency in young Xenopus tadpoles. AB - This paper investigates the proposal that the frequency of the swimming central pattern generator in young Xenopus tadpoles is partly determined by the population of glutamatergic premotor interneurons active on each cycle. During fictive swimming spinal neurons also receive cholinergic and electrotonic excitation from motoneurons. As frequency changes during swimming we make two predictions: first, since most motoneurons fire very reliably at all frequencies, the electrotonic and nicotinic drive from motoneurons should remain constant, and second, when swimming frequency decreases, the glutamatergic drive should decrease as the number of active premotor excitatory interneurons decreases. We have tested these predictions by measuring the excitatory synaptic drive to motoneurons as frequency changes during fictive swimming. The components of synaptic drive were revealed by the local microperfusion of strychnine together with different excitatory antagonists. After blocking the nicotinic acetylcholine receptor, the mainly glutamatergic excitatory synaptic drive still changed with frequency. However, when glutamate receptors or all chemical transmission was blocked, excitation did not change with frequency. Our predictions are confirmed, suggesting that premotor excitatory interneurons are a major factor in frequency control in the tadpole central pattern generator and that motoneurons provide a stable background excitation. PMID- 9861708 TI - Plasticity of the electric organ discharge waveform of the electric fish Brachyhypopomus pinnicaudatus. I. Quantification of day-night changes. AB - The electric organ discharge of the gymnotiform fish Brachyhypopomus pinnicaudatus is a biphasic waveform. The female's electric organ discharge is nearly symmetric but males produce a longer second phase than first phase. In this study, infrared-sensitive video cameras monitored the position of unrestrained fish, facilitating precise measurement of electric organ discharge duration and amplitude every 2 h for 24 h. Males (n = 27) increased electric organ discharge duration by 37 +/- 12% and amplitude by 24 +/- 9% at night and decreased it during the day. In contrast, females (n = 8) exhibited only minor electric organ discharge variation over time. Most of a male's increase occurred rapidly within the first 2-3 h of darkness. Electric organ discharge values gradually diminished during the second half of the dark period and into the next morning. Modulation of the second phase of the biphasic electric organ discharge produced most of the duration change in males, but both phases changed amplitude by similar amounts. Turning the lights off at mid-day triggered an immediate increase in electric organ discharge, suggesting modification of existing ion channels in the electric organ, rather than altered genomic expression. Exaggeration of electric organ discharge sex differences implies a social function. Daily reduction of duration and amplitude may reduce predation risk or energy expenditure. PMID- 9861709 TI - Effects of biparental rearing on ultrasonic vocalization (USV) responses of rat pups (Rattus norvegicus). AB - This study investigated the influence of social rearing on ultrasonic vocalization (USV) responses of 11- to 12-day-old rat (Rattus norvegicus) pups in isolation to the presence or removal of an anesthetized adult. Pups were reared with the dam or dam plus a virgin female (aunt), their biological sire, or a castrated male. All pups reduced rates of USV in contact with anesthetized adults. Pups' USV rates after the adult's removal depended on their rearing experience. After removal of the dam, pups increased USV rates over baseline (potentiation). Pups reared with aunts potentiated USV after contact with adult females but suppressed USV after contact with males. Pups reared with sires potentiated USV after contact with sires or strange males. Pups reared with castrated males did not suppress USV after contact with castrates but did after contact with unfamiliar intact males. Thus, pups' differential USV responses to contact with passive adults in isolation can be acquired during prior rearing experience. PMID- 9861710 TI - Hearing in a megachiropteran fruit bat (Rousettus aegyptiacus). AB - The Egyptian fruit bat (Rousettus aegyptiacus) is one of the few megachiropteran bats capable of echolocation. However, it uses rudimentary tongue clicks rather than laryngeally produced echo calls. We determined the audiogram of 2 bats using a conditioned avoidance procedure with fruit puree reward. At an intensity of 60 dB sound pressure level, the bats' hearing extended from 2.25 kHz to 64 kHz, with a region of good sensitivity between 8 kHz and 45 kHz. A dip in sensitivity at 32 kHz appears to be due to pinna directionality. The hearing of Egyptian fruit bats is typical for a mammal of that size and is not as limited as previously reported. Methodological issues, specifically training an animal to listen for low-intensity signals and imposing a significant cost for failing to report signals (i.e., misses), are discussed as the basis for the discrepancy between our results and earlier reports. PMID- 9861711 TI - Anticipating load torques produced by voluntary movements. AB - The stability of an object held between the finger and thumb depends on friction developed by grip force, normal to the contact surfaces, to overcome tangential load force. Previous research has shown that in lifting an object, grip force rises with the increase in gravitational load force as the hand takes the weight and that in moving an object, grip force is adjusted to meet movement-induced inertial load force. Those results demonstrated the anticipatory nature of coordination of grip force with load force. Whether grip force anticipates load torque was studied in this research. When participants were constrained to use grasp points where the grasp axis was manifestly distant from object center of mass, it was found that they made grip force adjustments in anticipation of load torques that tended to destabilize an object as a result of lifting or moving it. These adjustments imply use of information about object center of mass in movement planning. PMID- 9861712 TI - Syllabic effects in word processing: evidence from the structural induction paradigm. AB - Spoken words have a rich structural organization in memory, consisting of syllabic and subsyllabic representations. A phoneme monitoring paradigm, in which the target phoneme occurs more frequently in one syllabic position than another (e.g., onset of the 2nd syllable vs. the coda of the 1st syllable: neu-tral vs. nut-meg; C. Pallier, N. Sebastian-Galles, T. Felguera, A. Christophe, & J. Mehler, 1993) was used to explore the formation of syllabic structure during word processing. Experiment 2 investigated how a recognition system that uses syllabic structure processes words with unclear syllable boundaries (e.g., pa-lace or pal ace?). Two methodological issues were explored: The importance of a baseline condition for measuring effects of induction (Experiment 1) and the form of the representation used in the induction paradigm (Experiment 3). Findings suggest that syllabic structure begins to form early in word processing, and they demonstrate the adequacy of the induction procedure for measuring such processes. PMID- 9861713 TI - Reading Finnish compound words: eye fixations are affected by component morphemes. AB - The role of morphemic processing in reading was investigated in 2 experiments in which participants read sentences as their eye movements were monitored. The target words were 2-morpheme Finnish compound words. In Experiment 1, the length of the component morphemes was varied and word length was held constant, and in Experiment 2, the uniqueness of the initial morpheme was varied and the rated familiarity and length of the word were held constant. The length of the initial morpheme influenced the location of the second fixation on the target word and the pattern of fixation durations (although it had a negligible influence on the gaze duration of the word). The frequency of the initial morpheme influenced the duration of the first fixation on the target word, had a substantial effect on the gaze duration, and also influenced the location of the first and second fixations on the target word. Subsidiary analyses indicated that these effects were unlikely to stem from orthographic factors such as bigram frequency. PMID- 9861714 TI - Mechanisms of perceptual organization and auditory selective attention: the role of pattern structure. AB - Five experiments were conducted to determine whether unpracticed listeners shift auditory attention in an anticipatory fashion in accordance with the Frequency x Time structure of a rapid sequence of tones. Listeners were presented on each trial with a series of pure tones either ascending or descending in frequency in a predictable manner. In all experiments, judgments were made more quickly and accurately for targets that were inconsistent with pattern structure. Performance was shown to depend on the magnitude of the frequency violation, with better performance for larger violations. Further, the strength of this effect was attenuated when overall pattern predictability was reduced. These results suggest strongly that unpracticed listeners do not automatically allocate attention in accordance with pattern structure. Rather, it appears that a preattentive perceptual process acts to integrate information likely to have arisen from the same source and that this early perceptual processing imposes constraints on selection. PMID- 9861715 TI - Levels of representation in visual word recognition: a dissociation between morphological and semantic processing. AB - A robust semantic priming effect typically occurs in visual word recognition if the prime is read before a response to the target. However, this effect is dramatically reduced if a letter search is performed on the prime prior to responding to the target. Three lexical decision experiments document the new observation that morphological priming is preserved following letter search on the prime. This dissociation between morphological and semantic priming following letter search can be understood in the context of an interactive activation framework. In addition, the implications of these results for connectionist and compound cue accounts of word recognition, as well as the issue of automaticity in word recognition, are discussed. PMID- 9861716 TI - Electrophysiological evidence for a postperceptual locus of suppression during the attentional blink. AB - When an observer detects a target in a rapid stream of visual stimuli, there is a brief period of time during which the detection of subsequent targets is impaired. In this study, event-related potentials (ERPs) were recorded from normal adult observers to determine whether this "attentional blink" reflects a suppression of perceptual processes or an impairment in postperceptual processes. No suppression was observed during the attentional blink interval for ERP components corresponding to sensory processing (the P1 and N1 components) or semantic analysis (the N400 component). However, complete suppression was observed for an ERP component that has been hypothesized to reflect the updating of working memory (the P3 component). Results indicate that the attentional blink reflects an impairment in a postperceptual stage of processing. PMID- 9861717 TI - Visually perceiving heading on circular and elliptical paths. AB - Five experiments addressed the perception of curvilinear heading under various conditions of optical flow. Perception of heading was unaffected by optical noise (Experiment 1) and was successful and equally accurate for flows generated by circular and elliptical paths of locomotion (Experiment 2). In Experiment 3, random perturbations of vector magnitudes in general curvilinear fields did not reduce the accuracy of perceived heading. When vector directions were randomly perturbed in Experiments 4 and 5, curvilinear heading perception was impaired. Discussion focuses on the inappropriateness of the vector normal hypothesis to the general curvilinear case, the importance of the pattern of relative vector directions, and the practical and theoretical significance of investigating different forms of noncanonical optical flow. PMID- 9861718 TI - Orthographic, phonological, and articulatory contributions to masked letter and word priming. AB - A series of experiments assessed masked priming for letters and words that are visually similar (SIM) and dissimilar (DIS) in upper- and lowercase formats. For letters, robust DIS priming was obtained in a naming task, but this priming did not extend to a variety of non-naming tasks. For words, robust DIS priming was obtained in both naming and non-naming tasks. SIM letter and word priming extended to all tasks, but the effects were generally small for letters. The restricted set of conditions for DIS letter priming suggests that this priming is mediated by phonological-articulatory processes, and the generality of DIS word priming argues that abstract orthographic codes mediate these effects. Consistent with this conclusion, priming between homophones (for both letters and words) was found in a naming task, but little word homophone priming was obtained in a lexical decision task. PMID- 9861719 TI - What is learned during automatization? II. Obligatory encoding of spatial location. AB - Six experiments addressed the encoding of location information during automatization, to test a critical prediction of the instance theory of automaticity (G. D. Logan, 1988). Subjects searched 1- or 2-word displays for members of a target category. Specific targets appeared in the same locations consistently throughout training, and then location changed at transfer. Sensitivity to changes in location were assessed with implicit and explicit memory tests. In both tests, sensitivity depended on the number of locations the words could occupy (2 vs. 16). Sensitivity varied with the number of words presented (1 vs. 2) in the implicit test, but not in the explicit test. The results suggest that subjects encoded the locations of the words during automatization, which confirms the predictions of the instance theory. PMID- 9861720 TI - Effects of masked stimuli on motor activation: behavioral and electrophysiological evidence. AB - Three experiments investigated the influence of unperceived events on response activation. Masked primers were presented before a target. On compatible trials, primes and targets were identical; on incompatible trials, opposite responses were assigned to them. Forced-choice performance indicated that prime identification was prevented by the masking procedure, but overt performance and motor activation as mirrored by the lateralized readiness potential (LRP) were systematically influenced by the prime. The direction of these effects was unexpected: Performance costs for compatible and performance benefits for incompatible trials were obtained relative to a neutral trial condition. The LRP revealed a sequential pattern of motor activation. A partial activation of the response corresponding to the prime was followed by a reverse activation pattern. It is argued that these effects primarily reflect an inhibition of the response initially triggered by the prime. PMID- 9861721 TI - Asymmetric relationships among perceptions of facial identity, emotion, and facial speech. AB - Effects of variation in an irrelevant stimulus dimension on judgments of faces with respect to a relevant dimension were investigated. Dimensions were identity, emotional expression, and facial speech. The irrelevant dimension was correlated with, constant, or orthogonal to the relevant one. Reaction times (RTs) were predicted to increase over these conditions to the extent that the relevant dimension could not be processed independently of the irrelevant one. RTs for identity judgments were independent of variation in expression or facial speech, but RTs for expression and facial speech judgments were influenced by identity variation. Facial speech perception was affected by identity even when variation in the mouth region was eliminated. Moreover, observers could judge speech faster for personally familiar faces than for unfamiliar faces. The results suggest asymmetric dependencies between different components of face perception. Identity is perceived independently of, but may exert an influence on, expression and facial speech analysis. PMID- 9861722 TI - Visual structure and the integration of form and color information. AB - Recent evidence challenges the view that attention acts on the outputs of early filters dedicated to processing motion, color, and orientation. Instead, "proto objects" specified by shading, depth, direction of lighting, and surface information are thought to provide input to attentional processing. These findings are extended here to the parsing of occlusion-based contours. Multicolored occlusion structures were briefly presented and illusory conjunctions measured. More illusory conjunctions were made to structures in which color was inconsistent with form information, a result that can be explained by a property of the visual system that biases the integration of color to be consistent with form. Results show that this constraint was based on global structural descriptions rather than the local information provided by T-junctions and collinearity. Together, these results offer a new tool for the study of the binding problem in vision. PMID- 9861723 TI - Effect of deviations from temporal expectations on tempo discrimination of isochronous tone sequences. AB - The effect of deviations from temporal expectations on tempo discrimination was studied in 3 experiments using isochronous auditory sequences. Temporal deviations consisted of advancing or delaying the onset of a comparison pattern relative to an "expected" onset, defined by an extension of the periodicity of a preceding standard pattern. An effect of onset condition was most apparent when responses to faster and slower comparison patterns were analyzed separately and onset conditions were mixed. Under these conditions, early onsets produced more "faster" judgments and lower thresholds for tempo increases, and late onsets produced more "slower" judgments and lower thresholds for tempo decreases. In another experiment, pattern tempo had a similar effect: Fast tempos led to lower thresholds for tempo increases and slow tempos led to lower thresholds for tempo decreases. Findings support oscillator-based approaches to time discrimination. PMID- 9861724 TI - Measurement and interpretation of Q0 and Q1 band property changes of two cationic metalloporphyrins upon binding with B-DNA: electronic MCD, CD, and optical absorption. AB - Room-temperature Q-band electronic MCD, CD, and optical spectra are reported for the first time for two free and nucleic acid-bound cationic metalloporphyrins. Metalloporphyrins are the high-symmetry (C4v or D4h), four-coordinate tetragonal type MP(X) [M = CuII and PtII; P(X) = meso-tetrakis(X-N-methylpyridyl)porphine; X = 2 or 4], and the nucleic acid is native, B-form calf thymus DNA (CT DNA). For intercalation system PtP(4)/CT DNA, large optical (lambda 0, epsilon max) and MCD (lambda peak, lambda trough, A(aj), A(aj)/D(aj), and delta[theta]Mp-t/epsilon max) band parameter shifts, as well as a single negative (-) induced CD peak for each of Q0 and Q1, were observed upon binding of the porphyrin to chiral DNA. The directions and magnitudes of these changes are comparable to those observed for the Soret (B0) band of this system. Decreases of MCD/optical ratio delta[theta]Mp t/epsilon max (varies; is directly proportional to A(aj)/D(aj)) of 30% (Q0) and 50% (Q1) upon intercalation indicate substantial reductions of the Q0[1Eu(a) (0,0), approximately 1a1u1 4eg1] and Q1[1Eu(a)(0,1), approximately 1a1u1 4eg1] excited state angular momenta, . It is of additional interest that intercalation leads to intensity cancellation of one of the four A-term lobes, the (+)lobe of the Q0 MCD (+)pseudo-A-term, which was also observed previously for intercalation systems PdP(4)/poly(G-C)2 and /CT DNA. Application of the CD sector method to the constituent x- and y-polarized porphyrin edtms, square root of D(aj), of the Q0 (edtms mu0x and mu0y) and Q1 (edtms mu1x and mu1y) CD bands leads to the conclusion that PtP(4) is symmetrically intercalated between adjacent GC base pairs, specifically at 5'GC3' sites, with each of two adjacent 4 N-methylpyridyl groups extending into each of the major and minor grooves. For outside binder CuP(2), small optical and MCD band parameter shifts and smaller, single positive (+) induced CD peaks are observed for Q0 and Q1 upon interaction with CT DNA, similar to what is found for B0. Little or no change in the MCD/optical ratio, delta[theta]Mp-t/epsilon max, informs that external binding has only small effects on the excited state angular momenta, , of these bands. The composite of MCD, CD, and optical spectra are consistent with CuP(2) binding to CT DNA by one or both of two competing external modes, i.e., by an AT specific, edge-on minor groove mode at 5'TA3' sites and/or a face-on major groove mode with high selectivity for 5'AT3' or 5'CG3' sites. For Q0 and Q1 of each of the CuP(2)/CT DNA and PtP(4)/CT DNA systems, we observe that the genuine MCD (+) A-terms of these free MP(X)s retain their (+) sign on becoming pseudo-A-terms upon binding to the low-symmetry, asymmetric DNA sites, and this indicates that the external and intercalative modes result in the 4eg splitting, or delta LUMO, being less than the magnitude of 1a1u-3a2u energy separation, or delta HOMO. PMID- 9861725 TI - A chemical approach to systematically designate the pyranopterin centers of molybdenum and tungsten enzymes and synthetic models. AB - The recent growth in the chemistry of the oxo-molybdenum enzymes has demonstrated the need for developing systematic methods for naming and abbreviating the novel pterin cofactors that bind to the metal ion via the sulfur atoms of an ene-1,2 dithiolate moiety. Historically, the term "molybdopterin" was coined to designate a special pterin that binds molybdenum and the molybdenum-bound form was termed the "molybdenum cofactor". However, recent studies have demonstrated that this novel pterin also binds tungsten. Furthermore, considerable variation has been found in the pterin entity itself. Taken together, these facts show that molybdenum- and tungsten-containing enzymes possess a family of cofactors rather than a single "molybdenum cofactor". This article proposes a unified methodology for describing these cofactors and their metal-free pterin units in light of recent results from protein crystallography. The various numbering schemes that have been used for this heterocycle are considered, as well as the IUPAC rules which are currently being used for related tricyclic compounds. A unified methodology for uniquely designating and abbreviating each cofactor is proposed. The available chemical and spectroscopic information on the pyranopterin entities that are present in the molybdenum and tungsten enzymes, the precursors to these centers, and synthetic pyranopterins are in part the basis of the systematic names and simplifying abbreviations. PMID- 9861726 TI - Isolation and characterization of vanadium bromoperoxidase from a marine macroalga, Ecklonia stolonifera. AB - The bromoperoxidase has been isolated from the marine brown alga, Ecklonia stolonifera (83 kDa) and has been characterized. Bromoperoxidase requires vanadium for enzyme activity as has been evidenced by EPR spectroscopy. The enzyme activity increased ca. 250% with the action of V5+ on the isolated enzyme, since more than 2/3 of the protein molecules were in the apo form. The increase in the enzyme activity was specific to V5+, while Fe2+, Fe3+, and Cu2+ inhibited the enzyme activity. This effect of V5+ addition was inhibited in phosphate buffer, probably because phosphate and vanadate compete for the active site. The bromoperoxidase exhibited a high thermostability (Tm = 68 degrees C) and a high stability in organic solvents (completely intact even in the presence of 50% methanol, ethanol and 1-propanol). PMID- 9861727 TI - Nanocrystals of magnesium and fluoride substituted hydroxyapatite. AB - Hydroxyapatite nanocrystals synthetized in the presence of different concentrations of magnesium and fluoride ions in solutions--1, 5 and 10 at.% have been submitted to a structural and chemical characterization. The syntheses were carried out in the presence of low molecular weight polyacrylic acid, which has been verified to inhibit hydroxyapatite crystallization. The polyelectrolyte is adsorbed into the crystals during the synthesis and provokes a reduction of the mean crystal sizes. The reduction is greater along the direction orthogonal to the c-axis, suggesting a preferential adsorption of the polyelectrolyte on the crystalline faces parallel to the c-axis. Both magnesium and fluoride can be incorporated into the hydroxyapatite structure. On the basis of the values of the lattice constants and of the magnesium relative content of the solid phase, it can be suggested that probably just a part of magnesium is substituted for calcium, the remainder being adsorbed on the crystal surface. However, magnesium destabilizes the apatitic structure favouring its thermal conversion into beta tricalcium phosphate, and displays an inhibiting effect on the crystallization of hydroxyapatite. This last effect is enhanced by the simultaneous presence of polyacrylic acid. Fluoride substitution for hydroxyl ions into hydroxyapatite structure induces a slight increase of the crystal sizes along the c-axis direction. The data indicate that the experimental approach can be successfully used to prepare nanoapatite with crystallinity, crystal dimensions, composition, structure and stability very close to those characteristics of biological apatites. PMID- 9861728 TI - Structural and biological characterization of a novel spermicidal vanadium(IV) complex: bis(pi-cyclopentadienyl)-N,N-diethyl dithiocarbamato vanadium(IV) tetrafluoro borate, [VCp2(DeDtc)](BF4). AB - In a systematic search for vanadocene complexes with sperm immobilizing activity as a new class of contraceptive agents, we identified V(eta 5-C5H5)2((C2H5)2 NCS2)(BF4) (=[VCp2(DeDtc)](BF4)) as the most potent and stable spermicidal compound. Here we report the detailed biologic and physicochemical characterization of this lead spermicidal compound by computer-assisted sperm analysis, electron paramagnetic resonance spectroscopy, electrochemistry, and X ray crystallography. [VCp2(DeDtc)](BF4) crystallized in the monoclinic space group P2(1)/c, with unit cell dimensions a = 7.0877(4) A, b = 22.2881(14) A, c = 11.8021(7) A, beta = 94.107(1) degree, V = 1859.6(2) A3. The final structure of [VCp2(DeDtc)](BF4) had an R factor of 0.0581 for 3191 independent reflections. The two sulfur atoms of the dithiocarbamate and centroids of the cyclopentadienyl rings in this vanadocene complex with unique contraceptive potential occupy four tetrahedral--like coordination sites about the central metal atom. PMID- 9861729 TI - Lead, nickel and vanadium in seafood: an exposure assessment for Kuwaiti consumers. AB - Human exposure to lead (Pb), nickel (Ni) and vanadium (V) through the ingestion of six species of fish and two species of shrimp in Kuwait are determined. The study on seafood consumed by the Kuwaiti residents was analyzed by conducting a survey in five districts of the country, namely, Kuwait city (Capital), Farwaniya, Jahra, Hawaly and Ahmedi. Samples consisting of fish and shrimp were purchased from the local fish market during November 1995 and June 1996. Based on the survey, three major factors were investigated: (i) Pb, Ni and V concentration in the fish of locally consumed fish and shrimp; (ii) daily dietary intake of these elements in humans through consumption of seafood; and (iii) characterizing potential health risks associated with the estimated daily intakes. The risk associated with Pb, Ni and V in seafood was estimated based on a Hazard Index (HI). Although the highest HI (using the 95th percentiles of the daily intake of the element as the dose of concern) was observed for zobaidy, hamoor and shrimp, it is always < 1. This indicated that no serious health threats are associated with oil-related elements in fish and shrimp. PMID- 9861730 TI - Saturation of ecosystems with toxic metals in Sudbury basin, Ontario, Canada. AB - Mining and resource recovery activities have not been kind to ecosystems in the Sudbury basin, Ontario. The combination of logging, smelting, fires and erosion resulted in an unusual anthropogenic ecosystem of denuded barren land with lifeless lakes, or a micro-desert. Since the 1970s, however, the concerted efforts made to reduce the emissions and rehabilitate parts of the degraded ecosystem have resulted in improvements in water quality, and recoveries in phytoplankton, zooplankton, zoobenthos and fish communities but have had little impact on toxic metal concentrations in many lakes. We show that most of the catchments in the Sudbury basin have become saturated with Cu and Ni, and some with Zn and Pb. It is estimated that mobilization of metals stored in soils and glacial overburden by surface runoff, groundwater drainage and wind re-working of tailings can sustain the high concentrations of Cu and Ni in many lakes for well over 1000 years. Strategies to immobilize the pollutant metals in the watershed rather than further emission controls may be required for dealing with high levels of toxic metals in surface waters of the saturated ecosystems. PMID- 9861731 TI - Radiocesium accumulation in edible wild mushrooms from coniferous forests around the Nuclear Centre of Mexico. AB - Cs-137 and K-40 have been determined in soil samples and in wild edible mushrooms from a forest ecosystem located at the Nuclear Centre of Mexico (NCM) and in several surrounding localities. The transfer factors for Cs-137 were studied in 21 mushroom species from 1993 to 1997. The Cs-137 and K-40 determinations were performed using a gamma spectrometer system of low level counting with a high purity germanium (HPGe) detector. The local mushroom species that were found to show higher Cs-137 transfer factors were Clavariadelphus truncatus, Cortinarius caerulescens, Gomphus floccosus and Lyophyllum decastes. The Cs-137 levels obtained at the NCM in some mushroom samples were slightly lower than those from surrounding localities indicating that the nuclear facility has not emitted Cs 137 to the atmosphere. PMID- 9861732 TI - On the analysis of iodine-129 and iodine-127 in environmental materials by accelerator mass spectrometry and ion chromatography. AB - Based on a review of literature about the abundances of 129I (T1/2 = 15.7 Ma) in the environment we show that there is a severe lack of knowledge, in particular about natural, pre-nuclear levels. Among the two analytical techniques which are sensitive enough to investigate 129I in environmental materials, namely radiochemical neutron activation analysis (RNAA) and accelerator mass spectrometry (AMS), only AMS is capable of covering the natural, pre-nuclear levels. Since such AMS measurements require chemical separation of iodine from the matrix, a wide variety of separation schemes are necessary for environmental analyses. We report here on such schemes for the analysis of soils, plants and soft tissue. They are applied exemplarily to analyses of soils from the vicinity of Chernobyl. For chemical separations prior to analysis, contamination control and blank analyses are essential. Here, we discuss quality control procedures in detail, both for RNAA and AMS. In the case of AMS we use ion-chromatography (IC) for the determination of stable iodine. The IC analysis is included in the separation schemes for environmental materials. First AMS-analyses of terrestrial biospheric materials demonstrate that natural environmental levels of 129I are lower than previously deduced from investigations using RNAA, but higher than expected from model calculations. AMS is capable of providing the missing knowledge about the radioecology of 129I. PMID- 9861733 TI - Organotin compounds in surface and pore waters of Ganga Plain in the Kanpur-Unnao industrial region, India. AB - Organotin compounds (OTCs) belong to those chemicals most toxic to the aquatic organisms which are deliberately introduced into the aquatic system through anthropogenic activities. Various species of organotin compounds were detected in surface and pore waters of the Ganga Plain in Kanpur-Unnao industrial region in pre-and post-Monsoon periods of 1995. The extraction of these compounds was performed using a method of direct aqueous phase in situ ethylation with sodium tetraethylborate (NaBEt4). After extraction into hexane, they were detected by GC MIP-AED. The water of this area is contaminated with dimethyltin (DMT), monobutyltin (MBT), dibutyltin (DBT) and tributyltin (TBT) compounds. Concentrations of these compounds in surface water of the pre-Monsoon period of 1995 range from 2.1 to 70.1 ng Sn/l for MBT, 1.7-101.1 for DBT and 2.9-19.8 for TBT, whereas in pore water; 9.7-23.5 ng Sn/l for MBT, 11.2-18.0 for DBT and 8.7 32.6 for TBT. However, in the post-Monsoon period of 1995, surface water shows considerable decrease in concentrations: DMT below detection-1.8 ng Sn/l, DBT 3.0 5.4, TBT 3.1-3.6 and MBT is below detection. This study is a preliminary documentation of water pollution by OTCs in the Kanpur-Unnao region of the Ganga Plain and suggests the necessity of further detailed OTCs studies in other regions of the Ganga Plain. PMID- 9861734 TI - The use of hexacyanoferrates in different forms to reduce radiocaesium contamination of animal products in Russia. AB - Hexacyanoferrates have been identified as highly effective radiocaesium binders which effectively reduce radiocaesium uptake and transfer to milk and meat. In Russia a hexacyanoferrate called ferrocyn has been produced for use as a countermeasure. In 1989-1992, experiments were undertaken in Russia to study the effectiveness of four different ferrocyn materials as 137Cs binders, their potential toxicity, effect on production rates of cow milk, effect on animal health and ease of implementation in routine agricultural practice. Four different ferrocyn delivery forms have been used: 98% pure powder, sustained release rumen boli (15% ferrocyn), salt licks (10% ferrocyn) and sawdust with 10% ferrocyn adsorbed (bifege). In initial experiments with different cows, sheep and pigs these four ferrocyn materials were effective in reducing radiocaesium transfer to animal products. Daily administration of ferrocyn powder at a rate of 3-5 g per cow reduced 137Cs transfer by up to 90% in milk. One single administration of three boli per cow (containing 30 g ferrocyn per boli) reduced 137Cs transfer by 50-75% for a period of 2 months. Salt licks containing 10% ferrocyn (0.22 kg ferrocyn per 2.2 kg briquette provided once) reduced transfer of 137Cs up to twofold for up to 10 days whilst bifege, given at a rate of 30-60 g day-1 (3-6 g day-1 ferrocyn), reduced 137Cs transfer by 90-95%. However, large scale application of these ferrocyn materials on collective and private farms in agricultural trials in 1994 resulted in a lower effectiveness. Therefore, in 1996 a comparative assessment of the application of the four ferrocyn forms was made under carefully controlled conditions. The results fully validated the previous experimental data, and showed the importance of meeting recommended procedures for treatment, particularly when hexacyanoferrates are administered on a day-to day basis. PMID- 9861735 TI - Squamous cell carcinoma associated with lupus vulgaris. AB - Squamous cell carcinomas are known to arise in certain chronic, scarring dermatoses and also to be associated with exposure to ultraviolet radiation. We now report a case arising in a plaque of lupus vulgaris, the patient having received radiation from a Finsen lamp as a child for a tuberculous abscess in that region. PMID- 9861736 TI - Itraconazole pulse therapy is effective in the treatment of Majocchi's granuloma: a clinical and pharmacokinetic evaluation and implications for possible effectiveness in tinea capitis. AB - Majocchi's granuloma is a folliculitic and perifolliculitic dermatophyte infection of the dermis, a site that is not generally colonized by fungi in immunocompetent individuals. Topical agents are usually ineffective therapeutically because of the deep location of the infection. The objective of this study was to determine the effectiveness of oral itraconazole. We also examined the pharmacokinetics of the drug in scalp hair during pulse therapy. This information would then be useful in determining the efficacy of itraconazole administered by means of intermittent pulse dosing in the treatment of tinea capitis. Seven patients (age range 25-75 years) were treated up to three times with itraconazole pulse therapy, 200 mg twice daily for 1 week, with 2 weeks off between pulses. Samples of scalp hair and plasma were also obtained to determine the pharmacokinetics of the drug at these two sites. All seven patients responded to therapy, clinical and mycological cure being achieved after one pulse (one patient), two pulses (three patients), or three pulses (three patients, each with toenail onychomycosis); none relapsed over a 6-18-month follow-up period. In all six patients who received two or more pulses of itraconazole, almost complete cure was observed before the second pulse, with full resolution within 2 weeks of its completion. Itraconazole was also detected in the hair after 1 week, and at concentrations 2.6-fold and 3.4-fold higher, respectively, after the second and third pulses. After the discontinuation of therapy, itraconazole was then detectable in the hair for 9 months, at least in a female patient who did not have her hair cut. Two pulses of oral itraconazole therapy thus appear to be effective in the treatment of Majocchi's granuloma, and it is possible that one pulse may be sufficient in some patients. These data suggest that itraconazole pulse therapy should be effective in the treatment of tinea capitis. PMID- 9861737 TI - Giant bladder diverticulum in Ehlers-Danlos syndrome type I causing outflow obstruction. AB - We describe a 16-year-old patient with Ehlers-Danlos syndrome (EDS) type I and recurrent urinary retention caused by giant bladder diverticulum and review the literature on this association. PMID- 9861738 TI - Non-traumatic rhabdomyolysis with gluteal ulcer. AB - The case is presented of a 34-year-old woman with rhabdomyolysis due to massive intake of nimetazepam, a benzodiazepine hypnotic. On admission, the patient had numerous blisters all over the body. One of the blisters in the gluteal region developed into a deep ulcer accompanied by muscle necrosis although it was not at a pressure point. The ulcer needed surgical intervention. Rhabdomyolysis is caused by the lysis and necrosis of muscle due to direct or indirect injury, high fever, ischaemia, hypoxia or enzyme defects. Release of myocyte components into the circulation then may induce major problems, especially acute renal failure associated with hypermyoglobinaemia. However, there have been few reports of cutaneous ulcer formation in non-traumatic rhabdomyolysis. PMID- 9861739 TI - Zosteriform cutaneous metastases from squamous cell carcinoma of the stump of an amputated arm. AB - The clinical appearance of skin metastases varies over a wide morphologic spectrum, cutaneous metastases mimicking herpes zoster being rare. We now report the case of an 83-year-old male with zosteriform cutaneous metastases secondary to a squamous cell carcinoma (SCC) which developed in the stump of an amputated arm. The pathogenesis is speculative, but in this case, the zosteriform distribution might well be explained by perineural lymphatic invasion and spread. PMID- 9861740 TI - Prostatic adenocarcinoma with cutaneous metastases overlying oestrogen-induced gynaecomastia. AB - Carcinoma of the prostate gland is the second most frequent malignancy in males, accounting for 17% of cancer in men; between a third and one-half of these patients will have distant metastases at onset, but rarely cutaneous. We now report a case of prostatic adenocarcinoma with such metastases involving the right nipple and periareolar skin, overlying an area of hormone-induced gynaecomastia. PMID- 9861741 TI - Atopic dermatitis apparently caused by type 2 CD8+ T cells in an AIDS patient. AB - A patient with atopic dermatitis and acquired immune deficiency syndrome (AIDS) had demonstrated blood eosinophilia, high levels of serum IgE and intracytoplasmic IL-4 in the CD8+ but not in the CD4+ cells, decreased numbers of CD4+ cells, and low levels of IL-2 in both CD4+ and CD8+ cells. These results, reflecting a predominant influence of type 2 CD8+ cells, suggest they were involved in the genesis exacerbation of the patient's atopic dermatitis. PMID- 9861742 TI - Sarcoidosis presenting as palmar erythema. AB - Sarcoidosis, a multisystem disease of unknown aetiology, is characterized by the formation of non-caseating granulomas which may involve any organ of the body. The commonest sites of predilection are the lungs, skin and lymph nodes. We now report a patient who presented with palmar erythema which on biopsy confirmed the presence of non-caseating granulomas and who responded to systemic corticosteroids. We are unaware of any previous report in the literature of sarcoidosis presenting in this way. PMID- 9861743 TI - Axillary acne agminata (lupus miliaris disseminatus faciei). AB - Acne agminata is a papulo-pustular eruption typically affecting the face of young adults and characterized histologically by the presence of caseating granulomata in the dermis. We now describe two adults who developed the condition in the axillae. PMID- 9861744 TI - Lichen sclerosus and lichen planus: a spectrum of disease? Report of two cases and review of the literature. AB - There has long been controversy concerning the relationship between lichen planus and lichen sclerosus. Whilst these two conditions are now considered distinct, there are shared clinical and pathological features. We now describe two patients with the cutaneous involvement of both lichen planus and lichen sclerosus, presenting a review of similar reported cases and discussing the implications for pathogenesis of these two diseases. Neither lichen planus (LP) nor lichen sclerosus (LS) are uncommon yet they have only infrequently been reported as coexisting. In his original description of LS, however, Hallopeau considered it to be a variant of LP and Gougerot has also commented on a possible common pathogenesis for the two conditions. Features which tend to support such as association include the distribution of the cutaneous lesions, histopathological features such as hydropic basal cell degeneration and a band-like lymphohistiocytic infiltrate in the dermis, and the reported association with autoimmune disease. We now report two patients in whom coexistent cutaneous LS and LP was confirmed histologically. PMID- 9861745 TI - Lichen planus pemphigoides: two case reports. AB - Lichen planus pemphigoides (LPP) is a rare condition with characteristic clinical, histological and immunopathological features, in which a lichenoid eruption is found in association with bullae on both involved and apparently normal skin. We present two patients to illustrate the diverse clinical manifestations of this disease. PMID- 9861746 TI - Onychomatricoma: an unusual cause of nail bleeding. PMID- 9861748 TI - Narrow-band UVB phototherapy: nine months' study in a New Zealand practice. PMID- 9861747 TI - A case of congenital scleredema. PMID- 9861749 TI - Thalidomide treatment for hypertrophic lupus erythematosus. PMID- 9861750 TI - Generalized morphoea after antitetanus vaccination. PMID- 9861751 TI - Terbinafine (Lamisil) therapy for Microsporum canis scalp ringworm. PMID- 9861752 TI - Petrified ears. PMID- 9861753 TI - Phenacetin or anything: what makes or made analgesics nephrotoxic? PMID- 9861754 TI - Surgical site infections. AB - The New CDC-definitions for surveillance of surgical site infections (1992) take into account 3 classes of surgical site infections (SSI): superficial and deep incisional SSI, and organ/space SSI. The most important host-related risk factors for development of SSI are advanced age, morbid obesity, disease severity, an ASA score > 2, prolonged preoperative hospital stay, and infection at distal sites. Microbial contamination of the surgical site occurs mainly during the surgical intervention. Although exogenous contamination may be of concern, especially in clean operations, most surgical site infections are caused by microorganisms of the patient's own commensal flora. SSI rates vary according to the type and duration of the surgical procedure and the skill of the surgeon. Proper surgical technique is the most important factor in the prevention of SSI. Modification of host risk factors should be attempted whenever possible. In addition, adequate protocols for antimicrobial prophylaxis with antibiotics should be followed. Surveillance of surgical site infections is probably beneficial for SSI prevention. PMID- 9861756 TI - Passive particle agglutination test for screening of Treponema Pallidum antibodies in blood bank routine. AB - In most laboratories and blood banks, the TP.PA test has replaced the TPHA test for the screening of Treponema pallidum antibodies. In this study we show that the 1:20 serum dilution should be used to discriminate between negative and positive samples instead of the recommended 1:40 dilution, thus increasing sensitivity without a significant loss of specificity. PMID- 9861755 TI - Analgesic nephropathy: clinical features and pathogenesis. Original data on the incidence and prevalence in a single renal unit. PMID- 9861757 TI - A patient with fulminant systemic vasculitis type polyarteritis nodosa and negative histology of small bowel infarction. AB - A patient with fatal polyarteritis nodosa is described. In spite of overwhelming clinical signs the diagnosis of PAN could not be confirmed by angiography nor histology of macroscopic clearly involved small bowel tissue. Histology of lower limb tissue after amputation confirmed the diagnosis. PMID- 9861758 TI - Serum parathyroid hormone and calcium during the first days after resection of a parathyroid adenoma. AB - Serum parathyroid hormone, calcium and phosphate were daily monitored in 12, 30 and 30 patients respectively during the early postoperative period after removal of a solitary parathyroid adenoma. In all patients PTH concentration dropped to very low values on the first postoperative day, whereafter a rapid recovery began. So the first day is the best time to evaluate the results of the intervention. Hypocalcemia was most frequent on the third day. After ten days 26 patients showed a normal calcemia, while 4 patients still had a mild hypocalcemia (8-8.8 mg/dl). These data may guide management of patients after parathyroid surgery. PMID- 9861759 TI - Bronchiolitis obliterans organising pneumonia. A report of 11 cases and a review of the literature. AB - The clinical syndrome "Bronchiolitis Obliterans Organising Pneumonia" (BOOP) has to be considered in patients with a flu-like illness since some weeks, fine crackles, and on chest X-ray bilateral patchy infiltrates. There is no response to antibiotics. BOOP is essentially idiopathic, but associations to other conditions exist. Lung function is often restrictive; biochemistry is not pathognomonic. BAL shows a mixed cellular pattern. The gold standard for pathologic diagnosis is open or thoracoscopic lung biopsy. However, a BOOP pattern or reaction is often seen on histologic specimens without the clinical radiologic features of the BOOP-entity. Therapy consists of corticosteroids, which have to be prescribed for a long time at a rather high dose. Recurrence is frequent, but prognosis is good. Evolution to respiratory insufficiency and death is rare and may occur in rapidly progressive BOOP. This study reports on 11 cases (6 males/5 females) of clinical-pathological BOOP-syndrome (mean age 58 yrs, range 17-73 yrs), with an unexpectedly high mortality rate of 36% (4 cases). The disease was idiopathic in 7, and was associated with intake of amiodarone (in 1), with past Mycoplasma pneumonia (in 1) and with connective tissue disease (in 2). There was a history of a flu-like syndrome, cough and dyspnea of a mean duration of 4 months (range 1 week to 8 months). Lung function was mostly restrictive or/and obstructive with a diffusing capacity ranging between 47 and 95% predicted; there was hypoxia in about half of the patients. Chest X-ray and computed tomography (CT) scan showed a patchy consolidation with linear opacities (unilateral in 4 patients, bilateral in 5) and/or a ground glass pattern (in 4 patients), and a focal pseudo-tumoral lesion (in 1). Bronchoalveolar lavage showed a variable pattern of mixed, or eosinophilic or neutrophilic alveolitis. Histologic diagnosis was based on open lung biopsy (in 3), on thoracoscopic biopsy (in 2), on transbronchial biopsy (in 2), on wedge resection of the nodular lesion (in 1) and on postmortem lung biopsy (in 3). One patient recovered spontaneously, 1 remained cured after resection of the focal lesion, 7 were treated with 16-125 mg methylprednisolone (of whom 3 had a temporary flare-up during tapering the corticosteroids and 2 died after 1 and 3 months due to infectious complications), 2 died due to rapidly progressive BOOP. PMID- 9861760 TI - Pulmonary Langerhans' cell granulomatosis (histiocytosis X): clinical analysis of 8 cases. AB - A detailed retrospective analysis of 8 cases has been set up to obtain an update of the clinical, radiological, diagnostic and therapeutical aspects of pulmonary Langerhans' cell granulomatosis (PLCG), previously called Histiocytosis X. This disease represents 2.8% of interstitial lung diseases (ILD) in a registration by pneumologists in Flanders. Seventy five% of our patients were active smokers. Cough, dyspnoe and constitutional symptoms were the main presenting symptoms. There was a 37.5% frequency of pneumothorax during the whole disease evolution and all these patients had to be treated with chemical or surgical pleurodesis. Spirometric pattern was variable but CO-transferfactor (TLCO) was significantly impaired in all our patients. Radiologically nodules and/or cystic lesions were found with preserved volumes. Open lung biopsy led to the diagnosis in 6 of the 8 cases. Transbronchial biopsies (TBB) were found to be not sensitive. Systemic staging in our group showed 50% of the patients to have a second focus of organ involvement. Treatment with corticosteroids and possibly immunosuppressives was without clear effect. Since 25% of our patients ultimately became candidate for lung transplantation extrapulmonary staging is prerogative and the high frequency of pleurodesis procedures in the past medical history of our patients has important implications since bilateral pleurodesis is still a contraindication for lung transplantation in some but not all institutions. PMID- 9861762 TI - The implications of hyperhomocysteinemia in a patient with type 1 diabetes. AB - We present a patient with type 1 diabetes mellitus, who had severe, progressive macrovascular disease despite an optimal control of modifiable risk factors. Screening for other risk factors revealed a markedly elevated plasma homocysteine level, that was secondary to pernicious anemia. The biological and clinical implications of this association are discussed. PMID- 9861761 TI - The prothrombin gene G20210A variant in an unselected thromboembolic population. A Belgian prospective clinical study. AB - The presence of the 20210A allele of the prothrombin gene has recently been shown to be a risk factor of venous thromboembolism, probably mediated through increased prothrombin levels. The aim of the study was to determine the frequency of the prothrombin 20210A allele in 193 consecutive unselected patients with venous thromboembolism and 100 healthy controls and to analyze the clinical profile associated with this new inherited thrombophilic factor. In agreement with previous reports, we found a frequency of 7.3% of heterozygous carriers of the 20210A allele among patients and 1% among controls. We confirm that plasma prothrombin levels are more elevated in the individuals bearing the prothrombin 20210A allele compared with those who do not. We did not find any relationship between the presence of the prothrombin 20210A allele and either a family history of thromboembolism, the rate of recurrences or the age at disease onset. However, the co-inheritance in the same individual of both prothrombin 20210A allele and factor V Leiden was associated with a significantly lower age at disease onset suggesting a synergistic contribution of both abnormalities. PMID- 9861763 TI - Irradiated mandible after teeth extractions: an experimental study on mini-pigs. PMID- 9861764 TI - Long-term behaviour of osteoblast-like human alveolar bone cell cultures. PMID- 9861766 TI - Radiographic features of surgical specimens of temporomandibular joint. PMID- 9861765 TI - Ultrastructural identification of Weibel-Palade bodies in human gingival blood vessels. PMID- 9861767 TI - Localization of the interleukin-1 beta (Il-1 beta) by immunohistochemistry in healthy and inflamed human gingiva. PMID- 9861768 TI - Presence of HPV 16 sequences in oral lichen planus lesions. PMID- 9861769 TI - Histochemical evaluation of alkaline phosphatase in compound odontoma: a study of plastic embedded undemineralized material. PMID- 9861770 TI - [The bactericidal activity of mouthwashes containing 0.10%, 0.12% and 0.20% chlorhexidine digluconate]. AB - Antiseptic activity of five mouthrinses containing 0.10%, 0.12% and 0.20% chlorhexidine, as well as aqueous chlorhexidine solutions at the same concentrations were determined under usual conditions (advocated dilutions by the manufacturers and short contact time) and according to French Pharmacopoeia recommendations. The three aqueous solutions and two mouthrinses containing 0.12% chlorhexidine were not antiseptic according to the fixed criteria. Bactericidal activities of the different dilutions of products were tested with an appropriate micromethod against nine-bacterial species involved in periodontal disease. Efficacity of mouthrinses were variable and were not agreed with the aqueous chlorhexidine-corresponding solutions: the mouthrinse containing 0.10% chlorhexidine reached 0.20% aqueous solution efficacy, and the mouthrinses containing 0.12% chlorhexidine were generally less active than the 0.12% aqueous chlorhexidine solution. Therefore, antibacterial activity of this type of product cannot be anticipated with the sole concentration factor, excipients playing essential part in the chlorhexidine activity. PMID- 9861771 TI - The physician-scientist really is an endangered species. PMID- 9861772 TI - There has been study after study. PMID- 9861776 TI - Brown and Tufts scientists among those to receive NIH funding for AIDS research. PMID- 9861777 TI - The J.I.M. interview. James M. Wilson, MD, PhD. AB - Simplicity of concept lies behind the widespread appeal of human gene therapy. A missing or defective gene is safely replaced by a gene that functions normally. Alternatively genes might be used in the laboratory to manufacture essential proteins and enzymes to create gene-based drugs. The idea of using genes to cure genetic diseases is equally appealing to researchers and the public at large precisely because it holds the promise of a real cure. However, as with so many things in clinical research, turning a simple idea into useful medicine has proved to be remarkably difficult. Nonetheless, the opportunity that gene therapy represents remains very real and, within the past decade, hundreds of scientists have become putative gene therapists. Research in gene therapy is booming in academia and industry, with mixed results. James M. Wilson directs the Institute for Human Gene Therapy at the University of Pennsylvania School of Medicine and is president of the 1500 member American Society of Gene Therapy. In an interview with Investigative Medicine, Wilson discusses the state of the art. PMID- 9861778 TI - Portal levels of the isoprostane F2 alpha-III, a marker of lipid peroxidation, do not correlate with increased portal pressure in cirrhotic patients. AB - BACKGROUND: Isoprostane F2 alpha-III (iPF2 alpha-III), a recently described member of a family of prostaglandin F2 alpha isomers and a biologically active end-product of lipid peroxidation, has been reported to increase portal pressure in cirrhotic rats. We found that its urinary levels were elevated in cirrhotic patients. METHODS: To investigate whether portal levels of iPF2 alpha-III were elevated in cirrhotic patients and whether there was a relationship between these levels and the portal pressure in the same patients, peripheral and portal plasma from cirrhotic patients (n = 18) undergoing elective transjugular intrahepatic portosystemic shunt and appropriate controls (n = 18) were assayed for iPF2 alpha III levels by using a gas chromatography/mass spectrometry assay. Portal pressure was measured in all cirrhotic patients. RESULTS: Cirrhotic patients had higher peripheral plasma levels of iPF2 alpha-III [78 (27-150) pg/mL] than controls [18(10-30)pg/mL] (P < 0.001). Portal iPF2 alpha-III levels were higher than plasma peripheral levels [129(50-375) pg/mL; P < 0.0001]. No correlation was found between peripheral and portal levels of iPF2 alpha-III (Rho = 0.17, P = 0.5). Portal levels of iPF2 alpha-III and portal pressure did not correlate (Rho = 0.17, P = 0.49). CONCLUSIONS: This study shows that peripheral and portal levels of iPF2 alpha-III, marker of in vivo lipid peroxidation, are elevated in liver cirrhosis. There is no correlation between iPF2 alpha-III portal levels and the portal pressure observed in these patients. These findings suggest that this biologically active isoprostane does not directly contribute to the portal hypertension observed in hepatic cirrhosis. PMID- 9861779 TI - Investigation of an outbreak of vancomycin-resistant Enterococcus faecium in a low prevalence university hospital. AB - BACKGROUND: Until 1995, there were no cases of vancomycin resistant enterococcus (VRE) identified at our university hospital. From May 1995 to August 1996, we investigated a cluster of 10 cases of phenotypic class Van B Enterococcus faecium. METHODS: Patients were matched with controls who were on the same unit for at least 7 days prior to the case developing VRE. Control patients were age and sex matched if possible, and had duration of hospitalization at least as long as the number of days it took the patient to become VRE positive. We analyzed 16 independent risk factors using Epi-info version 6. Environmental cultures were obtained in the MICU where 5 of the patients were located. All 10 patient isolates and environmental isolates were analyzed by pulsed field gel electrophoresis (PFGE). RESULTS: PFGE confirmed the genetic relatedness of all 10 patient isolates and environmental isolates. The VRE-positive group was more likely to be immunosuppressed and to have exposure to 3 physicians. In the MICU, significant, P < 0.05) risk factors for VRE were higher Apache scores, location adjacent to a VRE case, duration of vancomycin and amino-glycoside use, duration of invasive catheter use, and diarrhea. Among the VRE-positive environmental cultures was a blood pressure cuff wash that was used on several patients. CONCLUSION: We hypothesize that a VRE strain was introduced into our hospital environment and was spread by personnel or contaminated equipment. As a consequence of this study, a hospital-wide VRE policy was implemented. PMID- 9861780 TI - Pharmacological effects of propylthiouracil on corticosterone secretion in male rats. AB - BACKGROUND: We investigated the direct effects of propylthiouracil (PTU) on corticosterone secretion both in vivo and in vitro. METHODS: Male rats were divided into 4 groups and then injected subcutaneously with saline, PTU, PTU plus thyroxine (T4), or T4 once daily for 2 weeks. After 2 weeks, rats were decapitated or received adrenocorticotropic hormone (ACTH), intravenously. Zona fasciculata-reticularis (ZFR) cells from normal, saline-, PTU-, PTU plus T4-, or T4-treated rats were incubated with ACTH, forskolin, 8-Br-cAMP, deoxycorticosterone (DOC) +/- PTU (1, 2, or 5 mg/mL) at 37 degrees C for 2 hours. Corticosterone concentrations in plasma and cell media, and 3':5'-cyclic adenosine monophosphate (cAMP) production in ZFR cells were determined by radioimmunoassay. The effects of PTU on the activities of steroidogenic enzymes in ZFR cells were measured by the amounts of intermediate steroidal products separated by thin-layer chromatography. RESULTS: The basal and ACTH-stimulated levels of plasma corticosterone in PTU-treated rats were lower as compared to saline-treated animals. Both basal and ACTH-stimulated corticosterone secretion were inhibited by PTU > 2 mg/mL in rat ZFR cells. The cAMP production induced by forskolin was lower in PTU, PTU plus T4, or T4-treated rats than in saline treated animals. Chronic administration of PTU or PTU plus T4 inhibited the 3 beta-hydroxysteroid dehydrogenase, 21 beta-hydroxylase, and 11 beta-hydroxylase activities. Administration of PTU (1, 2, and 5 mg/mL) suppressed the basal, ACTH, 8-Br-cAMP, forskolin, and DOC-stimulated corticosterone secretion in rat ZFR cells. Likewise, PTU > 2 mg/mL inhibited the ACTH and 8-Br-cAMP-stimulated levels of intracellular cAMP in rat ZFR cells. CONCLUSIONS: These results suggest that PTU counteracts both basal and ACTH-induced adrenal steroidogenesis through their attenuation of the activity of 11 beta-hydroxylase and cAMP production in rat ZFR cells. PMID- 9861781 TI - Matrix-mesangial cell interaction modulates migration of macrophages. AB - BACKGROUND: Macrophages (Mos) have been demonstrated to play an important role in immune-mediated renal injury. Accumulation of macrophages in the mesangium has been reported to be a key event in the development of focal glomerulosclerosis. We hypothesized that mesangial cells (MCs) and matrix interaction may be a determinant for the migration of Mos into the mesangium. Therefore, we studied the effect of the interaction between matrix and MCs on the migration of Mos. METHODS: Mouse MCs were plated on Petri dishes coated either with buffer, collagen type I, III, IV, or Matrigel in media containing 1% fetal calf serum for 48 hours. Subsequently, supernatants were collected and stored. The effect of these supernatants (conditioned media) was evaluated on the migration of Mos across a filter in a modified Boyden chamber. RESULTS: Conditioned media from MCs grown on Matrigel (MC-Matrigel interaction products, MC-MGP) enhanced the migration of macrophages across a filter in a modified Boyden chamber when compared with conditioned media from MCs grown on plastic, collagen type I, type III, or type IV (MC-PP, MC-CI, MC-CIII, and MC-CIV). MC-MGP enhanced the migration of Mos in a dose dependent manner. Anti-MCP-1 antibodies attenuated (P < 0.05) the MC-MGP-induced Mo migration (MC-MGP, 16.8 +/- 2.5 vs MC-MGP + anti MCP-1 antibody, 6.5 +/- 1.2 migrated macrophages/field, n = 12). Anti-TGF-beta antibodies did not attenuate MC-MGP-induced Mo migration. MCs grown on Matrigel showed a 5-fold increase of MCP-1 mRNA when compared with cells grown on plastic or collagen type IV. CONCLUSIONS: The present study suggests that matrix components may modulate the migration of Mos. This effect of MC-matrix interaction on macrophage migration may be mediated through the generation of MCP 1. PMID- 9861782 TI - Apolipoprotein A-I complexed with phospholipid promotes hepatic lipoprotein and apolipoprotein secretion in the perfused hamster liver. AB - BACKGROUND: Apolipoprotein A-I within high density lipoprotein (HDL) plays a significant role in the process of reverse cholesterol transport from peripheral tissues to the liver. However, additional roles are not well defined for it in hepatic cholesterol metabolism. We have previously shown in the hamster that dietary cholesterol supplementation resulted in enhancement of apolipoprotein A-I (Apo A-I) in secreted nascent hepatic very low density lipoprotein (VLDL), suggesting that apolipoprotein A-I itself may play a role in hepatic lipoprotein secretion. METHODS: Using the isolated hamster liver with Apolipoprotein A-I perfusion, we then examined the hypothesis that Apo A-I alone or in association with phosphotidylcholine (PC) i.e., Apo A-I/PC as a HDL-like particle, has effects upon hepatic lipoprotein and bile secretion. Ultracentrifugation was performed on perfusate samples at 3 hours on control vs treated livers (Apo A I/PC, Apo A-I, or PC) to access lipid and protein concentration in VLDL, low density lipoprotein (LDL) and HDL. Four to thirty percent gradient SDS polyacrylamide electrophoresis (PAGE) and Western blot analysis were used on delipidated lipoprotein fractions and microsomes to assess apolipoproteins Apo B, A-I, II, and E. RESULTS: We found that perfusion of reconstituted HDL vesicles containing human apolipoprotein A-I and PC (Apo A-I/PC) 10 mg and 10 mg, respectively, in 22 mL for 3 hours into isolated hamster liver increased cholesterol (CH) and triglyceride (TG) components in secreted HDL; 45- and 6 fold, and in LDL; 15- and 2-fold, respectively. No significant changes occurred in VLDL or in biliary lipids. Concomitantly, Apo A-I/PC perfusion increased Apo E and Apo A-II and HDL and Apo B in LDL, while Apo E decreased in VLDL. Apo A-I/PC perfusion did not change the apolipoprotein content of hepatic microsomes of the perfused liver. Perfusion of apolipoprotein A-I (without PC) or PC (without apolipoprotein A-I) had none of these effects. CONCLUSION: These results indicate that the perfused discoidal apolipoprotein A-I/PC particle affects hepatic lipoprotein assembly and secretion, whereby both lipid and apolipoprotein components are enhanced in secreted HDL and LDL of hepatic origin. PMID- 9861784 TI - How to change as pharmacy changes. PMID- 9861783 TI - Antidepressants inhibit the glucocorticoid stimulation of thyrotropin releasing hormone expression in cultured hypothalamic neurons. AB - BACKGROUND: The pituitary thyroid axis is frequently effected in human depression possibly due to alteration in hypothalamic thyrotropin releasing hormone (TRH) secretion. Since clinical recovery is associated with normalization of thyroid function, the direct effect of antidepressants on TRH expression in a well established fetal rat hypothalamic neuronal culture system was investigated. METHODS: Fetal rat hypothalamic neurons (day 17) in culture were treated with different concentrations of antidepressants with or without glucocorticoids for 7 days following which TRH content was measured by radioimmunoassay (RIA). RESULTS: The results showed that Imipramine (IMIP), a tricyclic antidepressant (TCA), decreased the TRH content in a dose-dependent manner (from 80.7 +/- 4.9, at 10( 9) mol/L, to 14.1 +/- 0.6, at 10(-5) mol/L, fmol/well; P < 0.05). Desipramine (DESI), another tricyclic antidepressant, also decreased the TRH content (from 63.6 +/- 2.5, at 10(-9) mol/L, to 12.6 +/- 0.4, at 10(-5) mol/L, fmol/well; P < 0.05). Sertraline (SERT) and Fluoxetine (FLUO), serotonin selective reuptake inhibitors (SSRI), also decreased TRH content in a dose dependent manner (from 83.9 +/- 7.9, at 10(-10) mol/L, to 7.6 +/- 0.4, at 10(-5) mol/L, and from 41.66 +/- 2.5, at 10(-8) mol/L, to 17.54 +/- 0.92, at 10(-6) mol/L, fmol/well, respectively; both P < 0.05). We then tested the effect of these antidepressants on the Dex stimulation of TRH content. IMIP, DESIP and FLUO at 10(-6) mol/L reduced the TRH response to glucocorticoid stimulation (36.4 +/- 4.0, 56.6 +/- 2.4, 23.75 +/- 4.0, respectively vs 107 +/- 7.5 fmol/well; P < 0.05). CONCLUSION: This raises the possibility that the enhanced thyroid function in depression, which we postulate, may result in part from glucocorticoid stimulation of TRH gene expression, can be reversed by antidepressants through a direct effect on the TRH neuron. However, other mechanisms may need to be invoked in addition since basal TRH content was also reduced. PMID- 9861785 TI - Collaborative practice agreements between pharmacists and physicians. PMID- 9861786 TI - Marketing pharmaceutical care services. PMID- 9861787 TI - A model for improving medication use in home health care patients. AB - OBJECTIVES: (1) To develop a model for the identification and resolution of problems associated with suboptimal medication use in elderly patients receiving home health care; (2) To select the most important identifiable problems and develop structured procedures for their resolution. DESIGN: Expert panel review, problem selection, and development of a problem resolution model and guidelines. SETTING: Home health care. PARTICIPANTS: A panel with expertise in home health nursing, pharmacy, clinical pharmacology, gerontology, pharmacoepidemiology, and health services research. INTERVENTIONS: A list of potential problems associated with the most frequently used classes of drugs was compiled for review by the panel. Problems that were controversial or that could not be identified in the home care setting were excluded. Panel members individually ranked the remainder. Detailed procedures for identification and resolution of the 15 top-ranking problems were developed. MAIN OUTCOME MEASURES: Not applicable. RESULTS: Potential medication problems were defined by both drug use and symptoms or clinical signs associated with specific adverse effects, to ensure that clinically relevant problems would be identified. The model developed for problem assessment and resolution was centered on the drug utilization review (DUR) coordinator and the attending home health nurse. Following guidelines developed by the panel, the DUR coordinator advises the home health nurse about identified problems and how to resolve them. One of these practitioners, usually the nurse, then contacts the attending physician to explain their concerns, offer potential solutions, and request instructions. CONCLUSION: A potentially useful model for the identification and resolution of medication problems in the home health care setting was developed. This model is currently being evaluated in a randomized controlled trial. PMID- 9861788 TI - Sale of tobacco products in pharmacies: results and implications of an empirical study. AB - OBJECTIVE: To provide national-level data concerning the percentage of pharmacies selling tobacco products, examine relationships between selling practices and pharmacy characteristic variables, and explore perceptions of conflicts between tobacco-selling activity and professional and personal values and the potential effects of such conflicts. DESIGN, SETTING, PARTICIPANTS: Data were collected from a geographically stratified systematic random sample of 899 pharmacies. Multiple mailings were sent to the attention of the pharmacy manager. A random sample of nonrespondents was also contacted by telephone, urging participation. MAIN OUTCOME MEASURES: Whether the pharmacy currently sold cigarettes and/or smokeless tobacco products, and if so, whether these practices differed from what respondents' personal or professional values tell them to do. Scales designed to measure job satisfaction, job-induced tension, and propensity to leave were also included. RESULTS: Slightly more than half (50.5%) of the pharmacies sold cigarettes and 35.4% sold smokeless tobacco products. Independents were less likely than chain pharmacies to sell tobacco products. For those respondents working in pharmacies where tobacco products were sold, 47.6% responded that this practice differs from what their personal values tell them to do and 63.9% replied that this practice differs from what their professional values tell them to do. Even when controlling for pharmacy type, respondents working in pharmacies that sold tobacco products had significantly lower levels of global job satisfaction, higher levels of job-induced tension, and a higher propensity to leave than did respondents working in pharmacies that did not. CONCLUSION: Decision makers in pharmacies where tobacco products are still sold should take a serious look at the justification for the continued availability of tobacco products in an environment that has a goal of promoting health. PMID- 9861789 TI - Perceptions of pharmacists about adverse effects of corticosteroid therapy: focus on osteoporosis. AB - OBJECTIVE: To assess the perceptions of pharmacists regarding the adverse effects of corticosteroids, in particular corticosteroid-induced osteoporosis. DESIGN: Mailed survey of a random sample of pharmacists. SETTING: Richmond, Virginia. PARTICIPANTS: 350 community and hospital pharmacists. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Respondents' knowledge of adverse effects of corticosteroid therapy in men, premenopausal women, and postmenopausal women; the content of respondents' usual patient counseling for low- and high-dose therapy; and respondents' opinions of regimens for prevention of osteoporosis. RESULTS: Pharmacists associated gastritis, weight gain, and mood changes with corticosteroid use in a hypothetical 45-year-old man or 45-year-old premenopausal woman. For a hypothetical 65-year-old postmenopausal woman, pharmacists more frequently counseled about weight gain, osteoporosis, and gastritis. Patient counseling focused on these adverse effects for both low-dose (5 to 10 mg/day) and high-dose (> or = 30 mg/day) prednisone use. Osteoporosis was considered more likely in patients receiving high-dose corticosteroids on a long-term basis. CONCLUSION: Pharmacists responding to this survey frequently overlooked the association between low- and high-dose corticosteroid use and decreased bone density. Educational efforts are needed so that pharmacists can fulfill their potential for educating patients, monitoring corticosteroid therapy, and detecting drug-induced complications. PMID- 9861790 TI - Medication use during the perinatal period. AB - OBJECTIVE: To briefly describe the drug therapy administered during the perinatal period of pregnancy for common maternal and fetal complications, and to identify those agents that should not be used for these conditions. DATA SOURCES: References were obtained from an ongoing literature search of peer-reviewed obstetric and gynecologic journals and other selected medical and pharmacy journals available in the English language. Primary search vehicle was a weekly review of the tables of contents of nearly 1,300 medical journals provided by Reference Update (Institute of Scientific Information, Philadelphia). MEDLINE searches were also conducted using key terms for each subtopic. STUDY SELECTION: Specific references were selected for each topic based on the adequacy of their study design, patient population, and a recent publication date. Reviews were used if a large number of primary references would have been required to adequately describe the topic. DATA EXTRACTION: Most references reflected the current opinions expressed in the Educational (Technical) Bulletin and Committee Opinion series published by the American College of Obstetricians and Gynecologists. Recent, well-conducted studies that arrived at different conclusions were also included. DATA SYNTHESIS: Data obtained from each reference reflected the conclusions of the authors based on their research or an analysis of the research on others on the appropriate use of the drug(s) for the specific condition being treated. CONCLUSION: Drug therapy during the perinatal period is frequently required and can be beneficial for the mother, fetus, and newborn. Many complications previously associated with severe morbidity and mortality, such as infections, premature rupture of membranes, preterm labor, hypertension, maternal pain during labor, and postpartum hemorrhage, are now controlled with appropriate pharmacologic therapy. All health professionals who provide services to pregnant women should be knowledgeable in this drug therapy. PMID- 9861791 TI - Update on treatment of rheumatoid arthritis. AB - OBJECTIVE: To review current treatment of rheumatoid arthritis (RA), as well as recent advances. DATA SOURCES: MEDLINE search from 1990 to 1998 for human studies using search terms "rheumatoid arthritis"; "cyclooxygenase inhibitors" combined with "anti-inflammatory agents, nonsteroidal"; "tumor necrosis factor" limited to "antagonists and inhibitors"; "isoxazoles." DATA SYNTHESIS: RA is a chronic inflammatory disease characterized by symmetrical joint involvement, usually of the small joints of the hands and feet. Although the hallmark of the disease is inflammation of joints, other organ systems--including the eyes, blood vessels, lungs, and cardiopulmonary system--may also be involved. Treatment of RA requires both drug and non-drug approaches. Current drug therapy consists of combinations of nonsteroidal anti-inflammatory drugs (NSAIDs) and disease-modifying antirheumatic drugs (DMARDs). Corticosteroids are also used either for short-term treatment during initiation of therapy, in bursts during acute disease flares, or chronically in low doses. A number of promising new agents are in development. NSAIDs with preferential inhibition of cyclooxygenase II may offer a better safety profile than existing agents. Leflunomide and biological agents such as etanercept may provide benefit for patients who fail to achieve adequate response from conventional therapy. CONCLUSION: Traditional approaches to treatment of RA include NSAIDs combined with DMARDs. New agents just reaching the market represent important advances and have the potential to make a positive impact on treatment of RA. PMID- 9861793 TI - Pharmacists' liability into the year 2000. AB - OBJECTIVE: To educate the pharmacy community regarding areas of potential liability arising from the pharmacist's role in the changing health care delivery system. DATA SOURCES: Published cases (LEXIS and Westlaw), literature (NEXIS and Westlaw), and abstracts available through July 1998. CASE SELECTION AND DATA ABSTRACTION: Selected on the basis of the authors' objectives and the usefulness of the information for practicing pharmacists. DATA SYNTHESIS: As the pharmacist's role in the health care delivery system continues to change, so too does the pharmacist's exposure to liability. Although historically a pharmacist had no common law "duty to warn," new laws requiring a pharmacist to counsel patients, along with increased scrutiny by the media, have increased the pharmacist's potential for liability. An evaluation of recent case law indicates that state courts are increasingly willing to extend a pharmacist's duty to warn where the pharmacist has special knowledge of the patient or the patient's condition, contraindicated drug usage, where a prescription substantially exceeds the maximum safe dosage, or where a pharmacist fills or refills a prescription without physician authorization. A recent issue that may add to the list of potential liability concerns is the substitution of generically equivalent narrow therapeutic index drug products. CONCLUSION: The pharmacist's role in the ever changing health care delivery system, along with the public's increased scrutiny of pharmacists' dispensing practices, may inevitably extend a pharmacist's duty to warn a patient of potential problems related to prescription drug products. This trend is expected to continue into the new millennium. PMID- 9861792 TI - Hormone replacement therapy and breast cancer: revisiting the issues. AB - OBJECTIVE: To assess current ideas about the benefits and risks of estrogen and hormone replacement therapy (ERT/HRT) in postmenopausal women. DATA SOURCES: MEDLINE searches, supplemented by various texts, of the literature on HRT, ERT, and selective estrogen receptor modulators (SERMs): tamoxifen, toremifene, and raloxifene. DATA SYNTHESIS: HRT is primarily used for improving quality of life in women suffering from vasomotor symptoms associated with menopause. HRT is beneficial in postmenopausal women for preventing cardiovascular disease, osteoporosis, and Alzheimer's disease. Review of meta-analyses of clinical trials showed that ERT/HRT ever-users (patients who have ever used ERT/HRT) did not have an increased risk of breast cancer, but current users did have an increased risk, with some studies reporting increasing risk with duration of ERT. No relationship was found between dose or the addition of progestin to ERT and increased breast cancer risk. Overall breast cancer mortality rates associated with HRT were decreased in current users. In general, HRT does not increase the risk of breast cancer in women with a family history of the disease, compared with those without a family history. New HRT strategies that could potentially prevent breast cancer are now being developed. The SERMs tamoxifen and toremifene appear to have positive clinical effects on bone and serum lipids; they are currently being investigated for use as breast cancer chemopreventive agents. Raloxifene, a new SERM used for the prevention of osteoporosis, is an alternative for women who cannot tolerate HRT. Unfortunately, these SERMS have anti-estrogenic effects and thus cause vasomotor adverse effects such as hot flashes and vaginal dryness. In addition, SERMs do not protect against heart disease or prevent osteoporosis as well as does HRT. CONCLUSION: Presently, SERMs will not become first-line HRT, as the positive effects of ERT/HRT may outweigh any potentially increased risk of breast cancer. The development of new agents with pharmacodynamic profiles similar to that of ERT/HRT but lacking its adverse effects would be greatly beneficial for postmenopausal women. PMID- 9861794 TI - Costs of implementing pharmaceutical care in community pharmacies. AB - OBJECTIVE: To estimate the costs and benefits to community pharmacies of converting a traditional practice into one based on pharmaceutical care. SETTING: Community-based ambulatory care pharmacies. PRACTICE DESCRIPTION: Community pharmacy. PRACTICE INNOVATION: Pharmaceutical care. MAIN OUTCOME MEASURES: Costs incurred and revenues received. DESIGN: Twenty-five community pharmacies that had made the transition from traditional practice to one based on pharmaceutical care returned a survey providing data on the costs and revenues associated with the transition. RESULTS: Mean total cost of making the conversion for the 25 pharmacies was $36,207. The largest cost component associated with the transition was personnel, which had a mean cost of $16,512 per pharmacy. Mean revenues received for pharmaceutical care by these 25 pharmacies was $3,687, mainly for disease management services. Pharmacies that spent more on the conversions, and used brochures and physician detailing as well as consultants and franschises, tended to be more successful in generating revenues from pharmaceutical care. CONCLUSION: Most pharmacies that have made the conversion to pharmaceutical care have not experienced an increase in profits as a result of that conversion. More effort needs to be directed toward improving the flow of revenues obtained from providing pharmaceutical care. PMID- 9861795 TI - Setting health standards for the 21st century. AB - OBJECTIVES: To describe the nature, substance, and impact of health standards and to speculate on how they may change during the early years of the 21st century. SUMMARY: The health care system has experienced dramatic changes over the past three decades. Health standards now cover the spectrum from statutes to regulations to guidelines, as well as less compulsory forms, and address every aspect of health, including the availability, quality, and cost of services; the safety, efficacy, and quality of products; the adequacy of processes and analytic methods; and the value and cost-effectiveness of health technologies and services. Standards cover promotion, protection, and gatekeeping functions. They are established in response to specific needs and are driven by social and political agendas; social policy will drive standards development to a greater extent in the 21st than in the 20th century. Technology advances are stimulating new forms of standards, primarily gatekeeping standards. Standards raise the question of compliance. The trend is increasingly toward less compulsory, more discretionary standards, with a greater percentage being established by quasi official and voluntary bodies. CONCLUSION: 21st century health standards must balance the rights of access with protection from unsafe, ineffective, and poor quality products and services during a period when decisions must be made faster. They must be more flexible, enabling consumers to choose their level of protection. Such standards will pose the same concerns about adequacy, equity, and freedom of choice as health standards pose now. Whether we will use 21st century standards to improve and advance our health as well as systems, products, and services that contribute to health more effectively than we have in the 20th century is a question to ponder. PMID- 9861796 TI - Use of a paging system to improve medication self-management in patients with asthma. PMID- 9861797 TI - Monoclonal antibody approved for treatment of Crohn's disease. PMID- 9861798 TI - Needles for immunizations: select the right tool for the job. PMID- 9861799 TI - Electronic prescriptions. PMID- 9861800 TI - ECG of the month. Prematurity and destiny. Premature atrial impulses. PMID- 9861801 TI - Chemical peels. AB - A chemical peel is a procedure in which a topically applied wounding agent creates smooth, rejuvenated skin by way of an organized repair process. This article describes the indications, classifications, operative procedure, and complications of chemical resurfacing. In addition, alternatives to chemoexfoliation are discussed. PMID- 9861802 TI - Radiology case of the month. Painful right hip. Fibrous dysplasia of the intertrochanteric and subtrochanteric region of right femur. PMID- 9861803 TI - The journal 150 & 100 years ago. November 1848 and 1898. PMID- 9861804 TI - Latex hypersensitivity/glove component reaction as a contributing factor to osteomyelitis of the hand: a case report and review of the literature. AB - A patient with a known glove-related allergy/irritation has been diagnosed with osteomyelitis of the left ring finger. The patient's susceptibility to bone and soft tissue infection is due to an open skin fissure and an eczematoid dermatitis due to a work-related glove reaction. This article is presented to alert readers of the Journal of the Louisiana State Medical Society to the possibility of encountering this unusual complication of latex glove wear and to familiarize them with irritant and allergic contact dermatitis. This report is relevant as the incidence of occupational-related glove reactions has risen and continues to rise. PMID- 9861805 TI - Rapid recovery of a child with Guillain-Barre syndrome using intravenous immunoglobulins. AB - The purpose of this article is to acknowledge the success of using intravenous immunoglobulins in children with Guillain-Barre syndrome and to review the literature. A 17-year-old African-American young lady with Guillain-Barre syndrome of 11-days duration underwent intravenous immunoglobulin treatment. Within 6 days, the patient was ambulating and was showing significant signs of improvement. The use of intravenous immunoglobulins in children with Guillain Barre syndrome has been reported in only a few instances. PMID- 9861806 TI - Melanoma during pregnancy: a case report and review of the literature. AB - Approximately 40,300 patients are diagnosed with cutaneous melanoma annually in the United States with a disease-related mortality rate of approximately 7,300 per year. Although treatment strategies have been defined for cutaneous melanoma, therapeutic approaches for this hormonally sensitive tumor are difficult in the gravid patient. Epidemiologic studies suggest that there continues to be an increasing incidence of melanoma with an associated decrease in age at presentation for patients in the United States. These trends suggest that approximately 35% of women will be diagnosed with melanoma during their childbearing years. Hence, the perplexing challenge of melanoma management during pregnancy will perhaps increase over the next decade. This case report documents the occurrence of cutaneous melanoma in women during pregnancy and highlights key issues in the natural history of this disease. Detailed review of the current literature related to melanoma during pregnancy will provide insight into optimal therapeutic approaches. PMID- 9861807 TI - A blueprint for reducing turnover among nursing assistants: a Louisiana study. AB - For decades, various exposes and reports have painted an unflattering portrait of the nursing home industry across the nation. Nursing homes in Louisiana have endured their fair share of publicity and criticism. The industry in this state has been accused of being preoccupied with profits rather than quality resident care. And, while there is much debate as to the validity of this complaint, there is solid agreement that competent and stable nursing assistants are the key to quality resident care. Unfortunately, the annual turnover rate of these essential employees ranges from 50% to 400%, nationally. This research identified the factors most responsible for the rate of turnover of nursing assistants employed in Louisiana nursing homes. Based upon the results of this study, pay, benefits, workload, and employee-employer relations, are not related to turnover. The analysis revealed that only three issues are associated with turnover--the number of beds, the number of beds per registered nurse, and the number of beds per social service worker. The message is clear: nursing home administrators must be very careful in stretching such resources. The number of beds assigned to an RN, and, in particular, the number of beds per social service worker are management issues that, if overextended, risk the turnover of nursing assistants. PMID- 9861808 TI - Prevalence of asthma and asthma-like symptoms in an adult population sample from Verona. ECRHS Verona. European Community Respiratory Health Survey. AB - The aim of the study was to assess the prevalence of asthma and asthma-like symptoms in the general population of Verona. A screening questionnaire, with seven questions on respiratory symptoms, was sent by mail to 3000 randomly selected subjects, aged 20-44 yrs. After three subsequent mailings, nonresponders were interviewed by telephone. The overall response rate was 92%. At least one respiratory symptom was reported by 44% subjects. The prevalence of the most frequent symptoms differed according to the type of contact: early respondents (i.e. those who responded by mail) were more likely to report symptoms than late respondents (i.e. those who were interviewed by telephone). Wheezing was reported by 11.3% of males and 8.0% of females (p < 0.01). The allergic rhinitis and hay fever prevalence was 16.9% with no differences owing to sex or age. The prevalence of an "episode of asthma" (defined as a self-reported attack of asthma or treatment for asthma) and of "current asthma" (defined as a self-reported attack of asthma, treatment for asthma or wheezing other than due to a cold with dyspnoea in the last 12 months) was 4.1 and 4.7%, respectively. No differences were found on the basis of age and sex. The prevalence of asthma was greater in urban (5.0%) than in suburban (2.7%) areas (p < 0.05), where lower pollution levels were registered. The greater prevalence in urban areas suggests that some factors related to the urban environment could play a role in the development of asthma and asthma-like disorders. PMID- 9861809 TI - Causes of chronic persistent cough in adult patients: the results of a systematic management protocol. AB - Chronic persistent cough (CPC) is a common symptom generally caused by postnasal drip syndrome (PND), bronchial asthma (A), chronic bronchitis (CB), and gastro oesophageal reflux (GOR). The purpose of this study was to confirm the value of a testing protocol for determining the causes of CPC in adult patients and for evaluating the outcome of its specific therapy. Ninety-two patients with unexplained CPC were sent to our Department between January 1994 and June 1996. The mean (+/- SE) duration of cough was 32.7 (+/- 4.5) months. We studied these patients (number) by applying an anatomical protocol, according to which clinical evaluation they underwent: chest (92) and sinus (90) radiography, spirometry (92), methacholine inhalation challenge (88), skin prick tests (67), oesophagoscopy (28), prolonged oesophageal pH monitoring (14), and bronchoscopy (49), as needed. The results of the standardized specific therapy refer to 87 patients because 5 patients were lost to follow-up. Thus, CPC was due to: sinusitis or chronic rhinitis plus PND in 56% of patients, CB in 18%, A in 14%, GOR in 5%, PND and GOR in 6%, A and GOR in 1%. The cough went away in 79/87 patients after specific treatment, based on the diagnostic findings, giving a success rate of 91%. The results of the present study confirm previous findings indicating that one or more causes of chronic persistent cough can be found, and that an elevated success rate of therapy was reached when an anatomic diagnostic protocol was used. PMID- 9861810 TI - New frontiers in the management of obstructive sleep apnoea syndrome. AB - This paper is the text of a talk given at the European Respiratory Society meeting in Berlin in September 1997 in a symposium organized by the Sleep Disorders Working Group. It covers new treatments for obstructive sleep apnoea which are not established as standard treatments. Although postural treatment was proposed a long time ago, few studies have investigated its efficacy in clinical practice. However, in view of the data concerning postural sleep apnoea, it certainly deserves more consideration. Oral appliances appear to have become more popular as an alternative to continuous positive airway pressure (CPAP) when surgery is not desired or not desirable. A few controlled studies have been undertaken to establish its efficacy, which is less constant and less predictable than with CPAP. Other experimental approaches include nerve and muscle stimulation, for which preliminary results are promising. In the area of drug treatment, there is nothing new. Finally, the questions of what to do when treatment does not work and who should receive treatment are addressed. PMID- 9861811 TI - Upper airway imaging in sleep apnoea syndrome: clinical applications. AB - Symptoms of habitual snoring and excessive daytime sleepiness are extremely common in the general population, and have poor predictive value in identifying patients with "true" sleep-related disordered breathing. The upper airways are the main anatomical site responsible for snoring and sleep apnoea; therefore, their examination via different means has been quite extensively assessed. Clinical examination may point to severe micrognathia or retrognathia, grossly hypertrophied tonsils, obvious macroglossia, and oedema and inflammation of the uvula and soft palate. A recently proposed model is promising, but has not been validated independently yet. Endoscopic investigations have been performed in awake as well as in sleeping patients, with the pharynx in relaxed or active states; their predictive value remains poor, both for diagnostic purposes and for identifying patients that may benefit from surgery. Radiographic and magnetic resonance imaging techniques have permitted a detailed understanding of the process of narrowing and collapse of the upper airways. Unfortunately, these techniques do not perform any better than the ones previously cited as clinically efficient tools for diagnosis in the population of patients suspected of sleep related breathing disorders. In conclusion, clinical examination of the upper airways remains part of the clinical evaluation of patients suspected of sleep related disordered breathing. Other imaging techniques may be used for research purposes, but do not yet seem to be worth including in the routine assessment of this population. PMID- 9861812 TI - Implications of pathophysiology for management of the obstructive sleep apnoea syndrome. AB - The pathophysiology of obstructive sleep apnoea syndrome (OSAS) is complex and incompletely understood, but is principally based on an imbalance between the collapsing forces of the upper airway (UA) during inspiration and the counteracting dilating forces of the UA dilating muscles. A narrowed UA is very common among OSAS patients, which is usually due, in adults, to nonspecific factors such as fat deposition in the neck or abnormal bony morphology of the UA. Functional impairment of the UA dilating muscles is particularly important in the development of OSAS, and patients have a reduction in both tonic and phasic contraction of these muscles during sleep when compared to normal subjects. Arousal plays an important role in the termination of each apnoea, but may also contribute to the development of further apnoea because of a reduction in respiratory drive related to the hypocapnia which results from postapnoeic hyperventilation. A cyclical pattern of repetitive obstructive apnoeas may result. A better understanding of the integrated pathophysiology of obstructive sleep apnoea syndrome should help both in the choice of optimum therapy for each individual patient and also in the development of new therapeutic techniques. PMID- 9861813 TI - Idiopathic pulmonary haemosiderosis in an adult. AB - We report a very, rare case of idiopathic pulmonary haemosiderosis (IPH) occurring in a 68 yr old patient. Following alveolar haemorrhage onset, the diagnosis was obtained by a process of elimination, after clinical, endoscopic, cytohistological and laboratory investigations. The immunoglobulin G directed against myeloperoxidase antigen (C-ANCA), which was detected, can be regarded as indicative of an occult vasculitis to be followed up. Current therapy and perspectives for idiopathic pulmonary haemosiderosis patients are discussed herein. PMID- 9861814 TI - Long-term oxygen therapy on the island of Crete, Greece. AB - The aim of this study was to investigate the extent of use, the selection criteria for usage and the conditions under which long-term oxygen therapy (LTOT) is provided on the island of Crete, Greece. A total of 545 patients using oxygen therapy at home in Crete were found from the records held by local hospitals, health insurance offices and oxygen supply companies. After randomization, 100 patients were selected, and finally 79 patients (64 males, 15 females), with a mean age of 71 yrs, were contacted and completed a questionnaire. The majority of patients (n = 57, 72%) using LTOT had a diagnosis of chronic obstructive pulmonary disease (COPD). In 67 (85%) patients LTOT was prescribed by a chest physician. All patients used large oxygen cylinders. The duration of daily oxygen use was significantly longer in the group of patients prescribed it by chest physicians than in those who was prescribed it by doctors from other specialities (8.8 h versus 4.7 h, p < 0.05). Of the patients, 47 (60%) fulfilled the criteria for LTOT. However, only 33 (43%) had accurate written instructions on how to use their home oxygen. A significant number of patients (n = 24, 29%) were still smokers and 63 (80%) had had a follow-up assessment. Our results suggest that COPD is the major grounds for LTOT. The daily duration of the oxygen therapy was longer in the group prescribed it by chest physicians. It is a concern that difficulties in setting up oxygen cylinders resulted in some ineffective usage. Physicians should stress the advantages of O2 concentrators. PMID- 9861815 TI - Management of respiratory failure in cystic fibrosis. PMID- 9861816 TI - Modalities of ventilation in patients with human immunodeficiency virus infection. PMID- 9861817 TI - Modalities of ventilation in lung transplantation. PMID- 9861818 TI - Mechanical ventilation in elderly patients. AB - Elderly patients are increasingly opting for intensive care unit (ICU) treatment with mechanical ventilation (MV). The aim of this study was to review specific aspects of MV in the older elderly (80-yrs-old and older). We retrospectively studied all patients who underwent MV during a 2-year-period in our respiratory ICU. Older elderly were compared with younger patients. Of 478 patients admitted to our unit, 58 underwent endotracheal ventilation (ETV) and 243 noninvasive ventilation (NIV). At the time of admission, older elderly patients (n = 106) were more severely ill than the younger ones, according to simplified acute physiology scores (SAPS). The history of older patients was characterized by a lower frequency of neurological abnormalities, and in the ETV group, a lower frequency of days previously spent in hospital. NIV was performed more often in older elderly (64%) than in younger patients (47%) and for a shorter time-period (8 compared to 10 days). The ICU mortality rate was higher in the older patients than in the younger ones in the overall population admitted to the unit (38 compared to 12%) and in the NIV population (21 compared to 9%) but not in the ETV population (40 compared to 44%). Long-term survival (2 yrs) was low (12%) in the older patients admitted to the unit. Several studies from the literature were reviewed. All authors agreed that age alone should not be a criterion to exclude the older elderly from intensive care, regardless of whether they found them to have the same or a poorer prognosis than younger patients. Selection biases are rarely studied. Our results indicate that some selection procedures on admission to the intensive care unit and before endotracheal ventilation are present despite the absence of any selection policy in our department. This selection enables us to obtain results from older populations which are as good as those from nonselected younger populations. When its use is practicable, noninvasive ventilation is associated with less discomfort, fewer complications and better short-term results than is endotracheal ventilation. In all cases, the long-term prognosis is poor. PMID- 9861819 TI - Modalities of ventilation in neuromuscular disorders and thoracic deformities. PMID- 9861820 TI - Modalities of ventilation in obesity. AB - Obesity is nowadays the most frequently found health risk in the USA, where more than 1 in 3 adults have a weight > or = 20% over the ideal value. Obese patients are more prone to developing sleep apnoea syndrome and obesity hypoventilation syndrome as well as more frequent postoperative complications. Thus, acute and chronic respiratory failure episodes represent current presentations in clinical practice where noninvasive ventilation is very efficient and must be guided by polysomnographic data in order to decide on long-term respiratory treatment to avoid recurrence of acute on chronic decompensation. PMID- 9861821 TI - Home mechanical ventilation: demographics and user perspectives. PMID- 9861822 TI - Energy balance and intermediate metabolism in chronic lung disease. PMID- 9861823 TI - Body composition, skeletal muscle function, and exercise performance in chronic respiratory disease. PMID- 9861824 TI - Nutritional and metabolic support strategies. PMID- 9861825 TI - Experience with new autotitrating nasal continuous positive airway pressure machines. AB - Since the introduction of nasal continuous positive airway pressure (nCPAP) in the treatment of sleep apnoea, enormous technical improvements in nCPAP devices have been achieved. nCPAP has become the most effective and widely used standard therapy for obstructive sleep apnoea. Recently, machines that automatically titrate the effective nCPAP pressure have become available. The question now is, whether and how they work and what the benefits and limitations of such devices are. The algorithms that control the machines are quite different and so is their effectiveness. Even in devices that generally work well, there may be some principal limitations. The algorithm may be mislead by artefacts, especially leaks. Apnoeas and arousal may occur because the pressure increase is too slow or breathing disturbances reoccur after a pressure decrease. Autotitrating nasal continuous positive airway pressure may reduce the work required to determine effective pressure. It is a valuable treatment option for patients who need high and variable nasal continuous positive airway pressure. It should not be used as an unattended pressure titration tool because this will lead to a loss of quality in patient care and a reduced acceptance of nasal continuous positive airway pressure, as pressure determination is only a small part of the adaptation of the patient to nasal continuous positive airway pressure. PMID- 9861826 TI - How to improve compliance to nasal continuous positive airway pressure in sleep apnoea syndrome. PMID- 9861827 TI - Sleep disordered breathing and cardiomyopathy. PMID- 9861828 TI - The ISHAM resolution 1997. International Society for Human and Animal Mycology. PMID- 9861829 TI - Synthesis of fluorochromes and pigments in Malassezia furfur by use of tryptophan as the single nitrogen source. AB - A new minimal medium consisting only of L-tryptophan (L-Trp) and a lipid source induced formation of brown pigmentation only in the species Malassezia furfur, which diffuses into the agar. Strains of the species M. sympodialis and M. pachydermatis failed to grow on this medium. On mDixon medium, however, after replacement of peptone by L-Trp, growth of all three Malassezia species was achieved. Under these conditions pigment production was observed with all M. furfur strains tested, although the results for M. pachydermatis strains were inconsistent. M. sympodialis strains showed no pigment production. On the minimal medium pigmentogenesis was induced in M. furfur by only 0.01 g% tryptophan; the pH optimum was pH 5. In all M. furfur strains, alternative amino nitrogen sources given concurrently with Trp suppressed pigmentogenesis. Furthermore, there were differences in the optimal temperature among the individual M. furfur strains. CBS 7019, CBS 6000 and CBS 6001 failed to produce pigment at 37 degrees C. The extract of the culture exhibited remarkable fluorescence, and several indole derivatives with a broad spectrum of colours were detected. This finding may have an impact on the clinical appearance of pityriasis versicolor, a very common skin disease caused by lipophilic yeasts of the genus Malassezia. We hypothesize that in pityriasis versicolor metabolic adaptation of Malassezia yeasts to altered nitrogen conditions on superficial skin might be of patho-physiological importance. Tryptophan as an inducer of pigmentogenesis probably accumulates during excessive sweating, a well-known manifestation of pityriasis versicolor. PMID- 9861830 TI - Anti-Malassezia furfur antibodies in the population. AB - The authors carried out research into anti-Malassezia precipitating antibodies in the population, specifying the distribution by age and sex. A total of 868 serum specimens from subjects of both sexes, aged between 0 and 80 years, were studied. An immunoelectroprecipitation reaction was used using a M. furfur culture filtrate as antigen. No antibodies were found in children under 11 years, whereas they were present after that age and reached maximum frequency in subjects between 31 and 40 years of age. In subjects up to 50 years of age they were more frequent in women, but in subjects over the age of 50 years they were more frequent in men. Globally, antibodies were found in 31% (270/868) of the subjects examined. The presence of antibodies correlates with data from the literature regarding the isolation of this yeast in relation to age. PMID- 9861831 TI - Fungaemia due to Cryptococcus laurentii and a review of non-neoformans cryptococcaemia. AB - Cryptococcus laurentii is one of several non-neoformans cryptococci that have rarely been associated with human infection. The spectrum of clinical infection due to non-neoformans species ranges from skin lesions to fungaemia. Most cases of non-neoformans fungaemia have been nosocomially acquired and have been associated with indwelling intravascular catheters and neutropenia. Limited data on in vitro susceptibilities of non-neoformans cryptococci show these species to be more resistant to fluconazole and flucytosine than most Cr. neoformans. Two such cases are presented here. PMID- 9861832 TI - Paranasal sinus mycoses in north India. AB - Recognizing the high incidence of paranasal sinus mycoses in north India, we analysed retrospectively the clinical, mycological and management aspects of 178 patients with proven disease attending our institute. On the basis of clinical, radiological, histopathological and mycological findings, the patients could be categorized into those with allergic (8), non-invasive (92) and invasive (78) disease types. Bony erosion without mucosal invasion by fungi was seen in 16 patients with non-invasive disease. Young men from rural areas were the most commonly affected. Rhinorrhoea with nasal polyposis (45.8%) and proptosis (46.4%) was the most common presentation. Concurrent involvement of the maxillary and ethmoid sinuses was common in these patients, whereas isolated sphenoid and frontal sinuses were involved in the invasive variety only. Orbital and intracranial extensions were detected in 100% and 13.2%, respectively, of patients with the invasive type of disease. Aspergillus flavus (79.7%) was the most common isolate. Surgical debridement and sinus ventilation were adequate for the effective management of the non-invasive disease. However, adjuvant medical therapy was included in treatment of the semi-invasive and invasive varieties of the disease. Itraconazole was found to be most useful in prevention of recurrence in the invasive type. Mortality was highest (33.3%) among patients with zygomycotic infection. Invasive fungal granuloma with orbital and intra-cranial invasion is a distinct entity in terms of its clinical course and treatment compared with non-invasive fungal sinusitis, and it needs to be treated aggressively with surgical excision and postoperative itraconazole. PMID- 9861833 TI - Therapy of sinuorbital aspergillosis with amphotericin B colloidal dispersion. AB - Invasive aspergillosis is a feared complication in the management of patients with malignancies. We report a 13-year-old boy with acute myelogenous leukaemia and chronic sinusitis who developed a sinuorbital fungal infection during cytostatic and prolonged antibiotic treatment. The clinical findings, diagnostic measures and treatment and its adverse effects are described and discussed. Special emphasis is given to our experience of the use of colloidal dispersed amphotericin B (Amphocil). PMID- 9861834 TI - African histoplasmosis: report of four cases from north-eastern Nigeria. AB - Four cases of African histoplasmosis with lesions of the skin, subcutaneous tissues and lymph nodes diagnosed histologically are described from the Borno State of Nigeria. Three of the cases were autochthonous to this State, whereas the fourth one originated from the Anambra State of Nigeria. These cases constitute the first record of African histoplasmosis from the arid zone of the north-eastern part of Nigeria. PMID- 9861835 TI - Systemic candidosis and concomitant aspergillosis and zygomycosis in two Amazon parakeets (Amazona aestiva). AB - Systemic candidosis and concomitant aspergillosis and zygomycosis were diagnosed immunohistochemically in two Amazon parakeets (Amazona aestiva). In the bird with systemic candidosis, subacute necrotic lesions were present in the lung and the gastrointestinal tract, whereas chronic giant cell-containing granulomas were located in the liver, heart, spleen and on the serosal lining of the small intestine. Although the lesions in the liver, heart and spleen most likely developed as a result of haematogenous spread, the granulomas on the serosal surface may have developed after a local transmural intestinal invasion. In the second bird, aspergillosis and zygomycosis were restricted to the lung, whereas some zygomycetes were found in the air sacs as well as in the heart and kidneys. In all organs the zygomycotic lesions were dominated by thrombosing vasculitis, supporting haematogenous dissemination. PMID- 9861836 TI - Humoral immunosuppressant activity of aflatoxin ingestion in rabbits measured by response to Mycobacterium bovis antigens using enzyme-linked immunosorbent assay and serum protein electrophoresis. AB - A total of 30 New Zealand white rabbits were divided into five equal groups. Animals in groups 1 and 3 were sensitized with bacillus Calmette-Guerin (BCG), and those in groups 2 and 4 with inactivated cells of Mycobacterium bovis (Sensitinogen). Group 1 and 2 rabbits were fed 2 ppm day-1 aflatoxin for 3 months. Group 5 served as control. Serum samples from animals in all groups were subjected to enzyme-linked immunosorbent assays (ELISAs) to determine antibody titre and to protein electrophoresis to determine immunoglobulin levels. The antibody titres and the immunoglobulin levels were significantly decreased in the aflatoxin-treated groups. PMID- 9861838 TI - HIV protease inhibitors influence the prevalence of oral candidosis in HIV infected patients: a 2-year study. AB - The introduction of HIV protease inhibitors was accompanied by reduction in HIV associated opportunistic infections. Therefore, we performed a retrospective study of HIV-infected patients to evaluate the effects of therapy with an HIV protease inhibitor (PI) on oral candidosis. This was of special interest, because an important virulence factor of Candida albicans is the secreted aspartic protease (SAP), which is assigned to the same class of aspartic proteases as HIV protease. Sixty-two patients were examined five times over a period of 2 years. There was a hint at a difference in the frequencies of C. albicans carrier state and manifest oral candidosis in favour of treatment with a PI. In addition, loss of Candida colonization and manifest oral candidosis was observed only in patients with elevation of CD4 cells upon PI. This might explain the effect, which also might go back to a direct inhibition of yeast SAP. PMID- 9861839 TI - Fungal colonization of the stomach and its clinical relevance. AB - The aim of this study was to estimate the frequency of fungal colonization of the stomach of patients suffering from gastric ulcer (GU) and chronic gastritis (CG) and the influence of fungal colonization of the stomach on the process of ulcer healing. We investigated 293 patients aged 20-80 years. Before and after 4 weeks of sucralfate treatment they underwent endoscopy of the stomach, histological examination of biopsies taken from the ulcer margin or inflamed gastric mucosa and mycological examinations of the gastric juice, surface brushing and biopsies. The studies revealed a high concentration of fungi in 54.2% patients with GU and 10.3% with CG. Candida albicans was the most frequently isolated organism. Fungal colonization of the stomach impairs the process of gastric ulcer healing. Control examination after 4 weeks of sucralfate therapy showed the ratio of GU healing in 62% of patients with a high concentration of fungi in comparison with 78% of patients not colonized with fungi (P < 0.05). A significantly longer duration of ulcer symptoms in the group of patients with a high concentration of fungi in the stomach was also observed. There was no correlation between the level of fungal antibodies, of Candida antigen in the serum and the concentration of fungi in the stomach. PMID- 9861837 TI - In vitro evaluation of voriconazole against clinical isolates of yeasts, moulds and dermatophytes in comparison with itraconazole, ketoconazole, amphotericin B and griseofulvin. AB - The in vitro activity of voriconazole (UK-109, 496), a new antifungal triazole derivative, against 650 clinical isolates of yeasts, moulds and dermatophytes was compared with that of itraconazole, ketoconazole, amphotericin B and griseofulvin. The geometric means of the minimum inhibitory concentrations (MICs) of voriconazole were 0.05 microgram ml-1 against yeasts (n = 187), 0.58 microgram ml-1 against moulds (n = 260) and 0.08 microgram ml-1 against dermatophytes (n = 203). The overall activity of voriconazole against yeasts and moulds was good, being similar to that of itraconazole, ketoconazole and amphotericin B. Voriconazole was highly effective against Aspergillus fumigatus (mean MIC 0.23 microgram ml-1) and other Aspergillus species and showed noteworthy activity (mean MICs 0.08-0.78 microgram ml-1) against emerging and less common clinical isolates of opportunistic moulds, such as Alternaria spp., Cladosporium spp., Acremonium spp., Chrysosporium spp. and Fusarium spp. On the other hand, voriconazole was less active in vitro than the comparative agents studied against various species of zygomycetes, such as Mucor spp., Rhizopus spp. and Absidia spp. Voriconazole and the other two azoles, itraconazole and ketoconazole, were more active than griseofulvin in vitro against most dermatophytes tested. PMID- 9861840 TI - Persistent oral candidosis by non-albicans Candida strains including Candida glabrata in a human immunodeficiency virus-infected patient observed over a period of 6 years. AB - A 38-year-old woman infected with human immunodeficiency virus (HIV) presented with persistent oral candidosis in which non-albicans Candida strains were the predominant yeasts in most of the examinations performed over a period of 6 years. Oral treatment with fluconazole had no effect on clinical signs of oral candidosis. In 8 of a total of 11 specimens, Candida glabrata, Candida parapsilosis and Candida tropicalis were at least suspected as the causative pathogens of oral candidosis. The non-response to fluconazole in our patient could be explained by in vitro resistance to fluconazole of detected Candida glabrata and Candida tropicalis isolates. PMID- 9861841 TI - Debilitating folliculitis barbae candidomycetica in a trumpeter: successful treatment with fluconazole. AB - A 29-year-old trumpeter, suffering from debilitating folliculitis barbae candidomycetica, was successfully treated with fluconazole 50 mg daily for a period of 4 weeks. Because of the strong local inflammation, oral treatment was initially combined with topical corticosteroid application. PMID- 9861842 TI - Oesophagitis caused by Candida kefyr. AB - The unusual case of an oesophagitis caused by Candida kefyr in a patient with squamous cell carcinoma of the oropharynx is reported. The further implementation of C. kefyr in the production of milk products is discussed. PMID- 9861843 TI - Tinea mammae mimicking atopic eczema. AB - Tinea mammae is a dermatophyte infection of the breast. Although rarely reported in this anatomical location, it can mimic other dermatoses. In this paper we describe an 85-year-old man with an erythema at the left breast which had enlarged over the previous 3 weeks. Trichophyton rubrum was isolated. Digital 20 MHz sonography revealed a reduction in echogenicity of the corium and a widened corium. Treatment with fluconazole over 5 weeks led to clearing of the lesion at the glabrous skin. At the end of therapy no difference between the involved and uninvolved skin areas was found sonographically. PMID- 9861844 TI - Onychomycosis due to Microsporum gypseum. AB - The first four cases of onychomycosis due to Microsporum gypseum observed in Italy between 1990 and 1997 are reported. clinical manifestation was distal subungual onychomycosis in all cases. The lesions were asymptomatic in two patients. Three patients were treated with oral itraconazole (pulsed therapy) and the other with ciclopirox 8% nail lacquer. Clinical and mycological recovery was achieved in all cases. The cases are reported because of their rarity. PMID- 9861845 TI - Cerebral blood flow and the response to acetazolamide during the acute, subacute, and chronic stages of aneurysmal subarachnoid hemorrhage. AB - Cerebral blood flow (CBF) and response to acetazolamide were measured during the acute, subacute, and chronic stages after aneurysmal subarachnoid hemorrhage and correlated with symptomatic vasospasm and clinical outcome in 45 patients who underwent early clipping of ruptured cerebral aneurysms, of whom 18 had symptomatic vasospasm and 27 did not. Xenon-enhanced computed tomography was used to measure CBF in both groups during the acute, subacute, and chronic stages, defined as days 0-4, 5-20, and > or = 21, respectively. Vasoresponse was assessed by the CBF increase in response to 1 g of acetazolamide administered after the baseline CBF study, except in the subacute stage of patients with symptomatic vasospasm. Outcome was scored based on activities of daily living 2-3 months after subarachnoid hemorrhage. CBF values and the response to acetazolamide were preserved during the acute stage but CBF values fell considerably below control values during the subacute stage in patients with vasospasm. The regions with flow values below 15 ml/100 g/min subsequently converted to infarction and the regions with those above 19 ml/100 g/min remained intact without infarction. During the chronic stage, low CBF persisted, but the response to acetazolamide was higher than that of the control group. Outcome scores were good and fair. CBF values were normal during all stages in patients without vasospasm. The response to acetazolamide fell transiently during the subacute stage. All outcome scores were excellent. In conclusion, the CBF informations soon after the onset of symptomatic vasospasm are useful to predict a reversibility of ischemic brain tissue and a final outcome. We suggest that vasospasm may cause a pathological or ischemic insult to brain tissue during the subacute stage, and the brain may remain metabolically depressed thereafter, leading to a poor outcome. Even clinically asymptomatic patients may suffer mildly vasospastic or ischemic conditions during the subacute stage. PMID- 9861846 TI - Effect of dexamethasone on cell proliferation of neuroepithelial tumor cell lines. AB - The effect of glucocorticoid on cell proliferation, the expression of glucocorticoid receptor, and the relationship between inhibition of cell growth and apoptosis were investigated in four established neuroepithelial tumor cell lines (KNS42, T98G, A172, and U251MG). Glucocorticoid receptor expression was located in the cytoplasm of untreated cells, but translocated into nuclei after treatment with dexamethasone in KNS42, T98G, and A172 cells. U251MG did not express glucocorticoid receptors. Dexamethasone significantly inhibited the growth of KNS42 and T98G cell lines, at high concentrations in contrast to growth stimulation at low concentration. Dexamethasone inhibited proliferation of A172 cell line at all concentrations from 10(-4) M to 10(-7) M. These were prevented by RU38486, a specific glucocorticoid antagonist. Apoptosis did not occur in any cell lines after dexamethasone treatment. There was no response to glucocorticoid by U251MG cells. Dexamethasone treatment of neuroepithelial tumor cells expressing glucocorticoid receptors causes translocation into the nucleus to modulate cell proliferation upon binding of different concentrations of dexamethasone in vitro. Dexamethasone inhibits proliferation of some neuroepithelial cell lines, not by glucocorticoid-induced apoptosis. The bimodal potential of glucocorticoid to stimulate or suppress proliferation of neuroepithelial tumor cells expressing glucocorticoid receptor must be considered in clinical trials. PMID- 9861847 TI - Cervical fast spin echo three-dimensional magnetic resonance myelography. AB - A technique of fast spin echo three-dimensional magnetic resonance (MR) myelography with flow compensation was developed for the evaluation of the cervical spinal lesions. The whole spinal cord and roots in the spinal canal can be visualized non-invasively on voxel images by the maximum intensity projection process to achieve the best static contrast of the cerebrospinal fluid. This method of MR myelography is applicable as a screening test for the patients with cervical spinal lesions. PMID- 9861848 TI - Cavernous angioma associated with venous angioma--two case reports. AB - A 17-year-old male and a 28-year-old female presented with cavernous angioma and venous angioma colocated in the same region, and manifesting as convulsions. The cavernous angiomas were extirpated without damage to the venous angioma. No postoperative neurological deficits were observed. The patients have been free of convulsions without anticonvulsant medication for 2 years. Early extirpation of cavernous angioma with presentation of venous angioma is recommended, preferably before bleeding occurs. PMID- 9861849 TI - Fatal epistaxis caused by rupture of an intratumoral aneurysm enclosed by a large prolactinoma--case report. AB - A 72-year-old female presented with episodes of epistaxis. Neuroimaging demonstrated a large prolactinoma totally enclosing a large intracavernous aneurysm of the internal carotid artery. Adjacent bony structures were eroded and destroyed by tumor invasion and extension. Rupture of the intratumoral aneurysm caused fatal epistaxis rather than subarachnoid hemorrhage before surgery. Intratumoral aneurysm is rare and epistaxis caused by rupture of it is extremely rare. Lack of bony protection apparently have contributed to the aneurysmal growth and rupture. PMID- 9861850 TI - Cerebral mycotic aneurysm treated with endovascular occlusion--case report. AB - A 56-year-old male with two mycotic aneurysms associated with infective endocarditis was treated by endovascular surgery before mitral valve replacement. Angiography revealed a ruptured proximal aneurysm and an unruptured distal aneurysm on the right middle cerebral artery. The ruptured aneurysm was successfully treated with an interlocking detachable coil, and patency of the parent artery was preserved. The unruptured distal aneurysm disappeared as a result of antibiotic therapy. Endovascular surgery of the mycotic aneurysm is less invasive and more effective than craniotomy under general anesthesia for patients with infective endocarditis. PMID- 9861851 TI - Transient neurological deficits simulating transient ischemic attacks in a patient with meningioma--case report. AB - A 67-year-old female had a history of transient neurological deficits involving fainting and right-sided hemiparesis. Magnetic resonance imaging showed a sphenoid ridge meningioma on the left, which had encased the internal carotid and middle cerebral arteries. Carotid angiography showed occlusion of the left internal carotid artery, a tumor stain, and engorgement of the surrounding cortical veins. Xenon-enhanced computed tomography showed reduced cerebral blood flow and poor response to acetazolamide in the surrounding brain tissue. The tumor was totally removed. Postoperatively, the patient had no more transient neurological deficits, and the response to acetazolamide was fully restored. An intracranial tumor may cause transient neurological deficits by reducing the cerebral perfusion pressure. This vascular insufficiency may occur when the tumor occludes major cerebral arteries, steals flow from the surrounding tissue, increases focal tissue pressure, and impedes regional venous outflow. The latter two factors were probably responsible in this patient. PMID- 9861852 TI - Symptomatic vessel narrowing caused by spontaneous rupture of craniopharyngioma cyst--case report. AB - A 36-year-old female presented with cerebral infarction due to severe vessel stenosis after spontaneous rupture of a craniopharyngioma, manifesting as aphasia and drowsiness. Neuroimaging showed the suprasellar cystic tumor with wall enhancement and cerebral infarction in the left temporoparietal region, and also enhancement of the left sylvian fissure and prepontine cistern. Angiography showed severe narrowing at the C1 portion of the left internal carotid artery (ICA) and the M1 portion of the left middle cerebral artery (MCA). The tumor was subtotally removed via a bifrontal craniotomy. There was accumulated milky-white debris around the left ICA and MCA. She became alert within a few days postoperatively. Repeat angiography 1 month after surgery demonstrated slight improvement of vessel narrowing. The neuroimaging and intraoperative findings suggested that the stenosis was due to vasospasm induced by chemical meningitis resulting from cyst rupture. PMID- 9861853 TI - Shock syndrome induced by bromocriptine test in a patient with prolactinoma--case report. AB - A 72-year-old male developed shock syndrome after a single dose of bromocriptine. He had undergone uncomplicated subtotal removal of an invasive prolactinoma in our department. The patient had normal ranges of pituitary hormones apart from hyperprolactinemia (167.7 ng/ml) after surgery. An acute suppression test with bromocriptine (2.5 mg per os) was done in the supine position 6 days following surgery. Three and a half hours after bromocriptine administration, he suddenly complained of anterior chest discomfort in bed. Cyanosis and profuse diaphoresis were noted. His blood pressure was 80/60 mmHg. Electrocardiography revealed sporadic premature contractions and slight depression in the ST segments. He recovered in about 10 hours after a rapid infusion of corticosteroid and lactic Ringer solution, and was discharged without sequelae. This is a very rare complication of bromocriptine, but the cardiovascular function of patients taking bromocriptine for therapeutic and diagnostic purpose should be monitored carefully. PMID- 9861854 TI - Rubella eradication: the countdown begins. PMID- 9861855 TI - Thirty years of medical teaching at the University of the West in Barbados. PMID- 9861856 TI - The role of euthanasia in the terminally ill. PMID- 9861857 TI - The role of euthanasia in the terminally ill. PMID- 9861858 TI - A profile of health research in Jamaica 1991-1995. Essential National Health Research Task Force. AB - A profile of health research in Jamaica between 1991 and 1995 was prepared in order to examine research capacity and needs as part of the process of promoting essential national health research. Of 43 organisations and research groups surveyed, 29 met the criterion of at least one peer reviewed publication between 1991 and 1995, and there were 201 health researchers. Most of the research groups had fewer than 20 professional staff with less than 10 engaged in health research. Institutional objectives and funding opportunities largely determined research priorities. 704 research papers were published over the period with 10 of the organisations responsible for 469 (66.6%). The number of research papers is overestimated because the same paper may be reported by more than one research group due to multiple authorship. On the whole, local research groups appear to be small, vulnerable, under-funded and deficient in basic equipment as well as trained and experienced researchers and support staff. These are compelling reasons for health researchers to come together to tackle common problems, promote collaboration and forge a joint strategy to strengthen health research capability in Jamaica. PMID- 9861859 TI - Surgical management of critical limb ischaemia in the French West Indies. AB - This retrospective study analyses the clinical presentation, surgical management and early outcome of 174 patients (mean age +/- SEM: 73 +/- 15 yrs) admitted for critical limb ischaemia. 145 (84%) had tissue loss at admission: toe gangrene or ischaemic ulcer in 77, and gangrene extending beyond the forefoot in 68. 87 primary limb amputations and 107 revascularisations were performed at iliofemoral (n = 20), suprapopliteal (n = 22) or infrapopliteal level. The postoperative mortality rate was 14% in the "Amputation" group and 9% in the "Revascularisation" group but the difference was not statistically significant. Infective complications were comparable in both groups, although 5 of 14 deaths after amputation were directly related to infection and all deaths after revascularisation resulted from cardiovascular complications. The early limb salvage rate after revascularisation was 82%. 19 secondary limb amputations were performed for bypass failure. Patients in whom primary amputations were required were older (p < 0.03) and had significantly higher rates of heart disease and nonambulatory status (respectively, 24 vs 17%, p < 0.05; and 37 vs 13%, p < 0.001) than patients in whom revascularisation was performed. Ischaemic rest pain and tissue loss confined to digit gangrene or ischaemic ulcer occurred more frequently than extensive gangrene in the "Revascularisation" group (p < 0.0001), while extensive gangrene extending beyond the forefoot occurred more frequently than ischaemic rest pain and tissue loss in the "Amputation" group (p < 0.0001). Late presentation of patients and enhanced tissue loss are probably the reasons for the higher primary amputation rate in our patients compared to that observed elsewhere. In patients amenable to revascularisation (56%), arterial reconstruction for critical limb ischaemia improves the chances of limb salvage. PMID- 9861860 TI - Quality of care of patients with diabetic foot problems in Barbados. AB - In Barbados diabetics with foot problems account for 80% of the patients in the female and 50% of those in the male general surgery wards, and many patients have major amputations for preventable problems. A six month prospective study was undertaken of all cases admitted with foot problems to the general surgical wards of the Queen Elizabeth Hospital (QEH) in order to determine the quality of foot care, particularly among diabetics. 67.5% of the 195 patients (55% female) admitted to the study were diabetic, most of whom were diagnosed 10 to 19 years previously. Most of the patients were 70 to 80 years old, but significantly more diabetics than non-diabetics were 40 to 70 years old. Foot problems in diabetics were precipitated by events that are considered trivial in non-diabetic patients. 87 (58%) of 150 responding patients had their feet inspected by health personnel in the previous year. 47 (63.5%) of the 74 who responded about the care of their nails said that they took care of their nails themselves. Nearly 40% of diabetic and non-diabetic patients had no reported source of care before their admission. 14 patients (10 diabetic) sought care the same day and 11 (nine diabetics) the day after noticing foot problems. Most patients presented with infection as part of their problem; recognition of the early signs of infection should be an integral part of the education of the diabetic patient. Educational efforts for patients must be continually reinforced because many patients said they had had no education about the care of their feet in the previous year. PMID- 9861861 TI - Impact of the prehospital trauma life support programme in Trinidad and Tobago. AB - The impact of the Prehospital Trauma Life Support (PHTLS) programme, introduced in Trinidad and Tobago in 1992, was assessed by questionnaires completed by 26 medical personnel (MP); 71 ambulance personnel (AP); and 50 non ambulance paramedical personnel (NAP). Of the 23 MP, 45 AP and 38 NAP who were aware of the programme, 19 (82.6%) MP, 40 (88.9%) AP and 25 (65.8%) NAP were able to differentiate personnel that had taken the PHTLS programme based on their performance. 32 (71.1%) of the AP were PHTLS trained. 24 (53.3%) and 4 (9%) of the AP identified poor equipment and poor supervision, respectively, as reasons for difficulty in applying PHTLS principles. Improvements observed among those completing the PHTLS programme were: improved resuscitation techniques by 20 (86.9%) MP, 38 (84.4%) AP and 27 (71.1%) NAP; better vital signs recording by 8 (34.8%) MP, 27 (60%) AP and 8 (21.1%) NAP; improved immobilization by 23 (100%) MP, 40 (88.9%) AP and 33 (86.8%) NAP; better haemorrhage control by 22 (95.6%) MP, 40 (88.9%) AP and 24 (63.2%) NAP; appropriate splinting of fractures by 23 (100%) MP, 40 (88.9%) AP and 32 (84.2%) NAP; and increased utilization of oxygen by 15 (65.2%) MP, 31 (68.9%) AP and 21 (55.3%) NAP. 32 (71.1%) AP with PHTLS training indicated improvement in their ability to resuscitate and transport trauma victims, with 42 (93.3%) reporting improvement in overall prehospital care. Medical, paramedical and ambulance personnel all perceive a significant positive impact of PHTLS training on prehospital trauma care. Although improvements in supervision, documentation and equipment are still required, improved trauma resuscitative techniques after PHTLS training should improve trauma patient outcome in Trinidad and Tobago. PMID- 9861862 TI - The seroprevalence of hepatitis and retroviral infection in Jamaican haemodialysis patients. AB - 63 haemodialysis (HD) patients and 63 age and gender matched controls were investigated for hepatitis B surface antigen (HbsAg) and antibodies to hepatitis B virus (anti-HBV), hepatitis C virus (anti-HCV), hepatitis D virus (anti-HDV), human immunodeficiency virus types 1 and 2 (anti-HIV-1 and 2) and human T-cell lymphotropic virus type-1 (anti-HTLV-1). The notable finding was an increase in hepatitis B markers, 34.9% in HD patients compared to 19.0% in controls (p < 0.02). The seroprevalence of anti-HCV (7.9%, p < 0.03) and anti-HTLV-1 (9.5%; p < 0.006) was also increased in the patients. Four of the five patients positive for anti-HCV were also seropositive for HBV. Anti-HIV and anti-HDV were not detectable in the HD patients in this study. The possibility of HTLV-1 being transmitted by organ transplantation is raised. The seropositivity rate for hepatitis B and C increased with duration on dialysis, but it is unlikely that it was related to the number of blood transfusions since 50% with no transfusion were HBV seropositive. PMID- 9861863 TI - Tumours and tumour-like lesions of bone. The UHWI experience. University Hospital of the West Indies. AB - A review of all primary tumours and tumour-like lesions of bone diagnosed in patients who attended the University Hospital of the West Indies over a 10-year period revealed 136 cases comprising 69 benign and 54 malignant tumours, and 13 cases of tumour-like lesions. The prevalence and clinical characteristics of the various lesions are discussed and compared with the findings in other series. The results of this analysis provide demographic data useful in the differential diagnosis of bone lesions locally. PMID- 9861864 TI - Psychiatric symptoms and an anterior cranial fossa meningioma. AB - We present a case of a patient admitted to a psychiatric hospital with psychotic symptoms and cognitive impairment but who was subsequently found to have an anterior interhemispheric falx meningioma. There must be a high index of suspicion for organic brain disease in patients over age 45 years presenting with psychotic symptoms and seizures for the first time. PMID- 9861865 TI - Tubo-ovarian abscess after tubal ligation. AB - Tubo-ovarian abscess usually results from ascending infection of the lower genital tract. In a few cases it can occur as a result of direct contamination at the time of tubal sterilization. We describe a case that presented seven years after post partum tubal sterilization, showing both acute and chronic components. PMID- 9861866 TI - From peptides to peptidomimetics: design of nonpeptide ligands for major histocompatibility proteins. AB - The ever increasing data available on antigen presentation by class I or class II histocompatibility proteins have made these glycoproteins highly interesting pharmaceutical targets for either vaccination or immunosuppressive therapy of autoimmune diseases and cancers. Herewith, we review the design and biological properties of the very first nonpeptide ligands of major histocompatibility proteins as well as their potential application in vaccination, Major Histocompatibility Complex (MHC) blockade or T cell receptor antagonism. PMID- 9861867 TI - Preparation of casein-chitosan microspheres containing diltiazem hydrochloride by an aqueous coacervation technique. AB - Sustained release casein-chitosan microspheres containing diltiazem hydrochloride (DTZ) were prepared with colloidal coacervation technique in a completely aqueous environment. The interaction between chitosan solution in dilute acetic acid (5% v/v) and casein solution in 0.5 M sodium hydroxide was the basis for the microspheres formation. Formaldehyde was used for the surface hardening of the droplets by cross-linking and thus fixing the shape and surface morphology of the formed microspheres. The entrapment efficiencies of the microspheres were variables (14.5-53.7%) depending on the preparation conditions. The prepared microspheres exhibited an angle of repose values between 31.9-42.0 degrees indicating good free flowing nature, whereas DTZ powder as such was non-flowable. The dissolution profiles of DTZ from casein-chitosan microspheres showed retarded release pattern of the drug into distilled water. Casein and chitosan concentrations, initial drug concentration and stirring time were found to be the main parameters that affect the properties and the performance of the prepared microspheres. The retarded release of DTZ was increased by increasing casein concentration, and stirring time. On the other hand, increasing chitosan concentration and using high initial drug loading showed a fast drug release. PMID- 9861868 TI - Pharmacodynamic and pharmacokinetic evaluation of lipid microspheres of indomethacin. AB - Lipid microspheres (LM) of indomethacin were prepared using phosphatidyl choline and soya bean oil. LM of required size (< 1 micron) were obtained by intermittent microscopic observation while homogenization. Anti-inflammatory activity of lipo indomethacin was compared with free indomethacin by carrageenan induced rat paw edema model. It was found that at 30% edema inhibitory dose, lipo-indomethacin was about 1.5 times more potent than free indomethacin, indicating possible localization of LM at the inflammatory site. Biodistribution studies were performed in rats at the dose of 12 mg/kg. After 2 h of the treatment, concentration of the drug was 0.31 microgram/g of inflammatory tissue with lipo indomethacin, whereas, only 0.05 microgram/g of the tissue was found with free indomethacin. Maximum difference in drug concentrations between the above two formulations was observed in lungs. LM were found not to cross the BBB as drug concentration in the brain after lipo-indomethacin treatment was below the detectable level. Thus, the pharmacokinetic data gave a quantitative evidence for high anti-inflammatory potential of lipo-indomethacin. PMID- 9861869 TI - Development of an abdominal magnetic resonance (MR) contrast agent formulation with Oral Magnetic Particles (OMP, Ferristene). AB - A formulation for fast dispersible granules with the magnetic resonance (MR) contrast agent, Oral Magnetic Particles (OMP), was developed. The formulation contained viscosity-increasing agents (xanthan gum and a mixture of microcrystalline cellulose and sodium carboxymethylcellulose). The granules were dispersed easily in water when stirred by hand and became a viscous OMP suspension with apparent homogeneity, but does not form granule aggregates (lump), within 10 min. The dispersibiility was greatly affected by the binder type and addition of disintegrant. Hydroxypropylcellulose and low-substituted hydroxypropylcellulose were preferable. The preparation method to incorporate OMP into the granules also greatly influenced the dispersibility of the granules. A fluidized bed granulation of the base granules containing additives and subsequent coating of OMP onto the granules was eventually utilized. The formulation was able to control the viscosity of the suspension by regulating the content of viscosity-increasing agent while maintaining dispersibility. The developed formulation gave a good MR contrast image in rats and no magnetic susceptibility artifact was observed. PMID- 9861870 TI - Synthesis of aryl semicarbazone of 4-aminoacetophenone and their anti-HIV activity. AB - The thioureido derivative of 4-aminoacetophenone aryl semicarbazone have been prepared. These derivatives have been characterised on the basis of different physicochemical evidences. The anti-HIV-1 (HTLV-IIIB) and -HIV-2 (ROD) activity and cytotoxicity of the compounds were tested. The compound VII and VIII showed maximum protection among the series. PMID- 9861871 TI - The family physician and international adoption. PMID- 9861872 TI - IUDs: time for a renaissance. PMID- 9861873 TI - Dealing with loss in multiple pregnancies. PMID- 9861874 TI - Pseudoneurologic symptoms in post-traumatic stress disorder. PMID- 9861875 TI - Radical prostatectomy in the treatment of prostate cancer. PMID- 9861876 TI - An office approach to the diagnosis of chronic cough. AB - Chronic cough is a common problem in patients who visit family physicians. The three most common causes of chronic cough in those who are referred to pulmonary specialists are postnasal drip, asthma and gastroesophageal reflux. The initial treatment of patients with cough is often empiric and may involve a trial of decongestants, bronchodilators or histamine H2 antagonists, as monotherapy or in combination. If a therapeutic trial is not successful, sequential diagnostic testing including chest radiograph, purified protein derivative test for tuberculosis, computed tomography of the sinuses, methacholine challenge test or barium swallow may be indicated. By using a standard protocol for diagnosis and treatment, 90 percent of patients with chronic cough can be managed successfully in the family physician's office. However, in some cases it may take three to five months to determine a diagnosis and effective treatment. For the minority of patients in whom this diagnostic approach is unsuccessful, consultation with a pulmonary specialist is appropriate. PMID- 9861877 TI - Primary care of international adoptees. AB - International adoptees are presenting to family physicians with increasing frequency. U.S. citizens have adopted over 100,000 international children since 1979. Prospective parents may seek advice from their physician during the adoptive process. If available at all, medical information on the child is often scanty. History and physical examination alone are often insufficient for diagnosis of common problems in this population. Adoptive parents may have concerns about growth and development, and appropriate immunizations. In addition, bacterial, viral and parasitic infections endemic in countries of origin create unusual challenges for the U.S. primary care physician. A basic understanding of the process of international adoption, a skillful evaluation of the child and selected laboratory studies enable the family physician to support the prospective parents and assist in a smooth transition of the child into a new family. PMID- 9861878 TI - Sarcoidosis: a primary care review. AB - Sarcoidosis is a multisystemic disorder of unknown etiology that most commonly affects adults between 20 and 40 years of age. Patients with sarcoidosis frequently present with bilateral hilar lymphadenopathy and pulmonary infiltration, and often with ocular and skin lesions. The diagnosis is established when clinical and radiographic findings are supported by histologic evidence of non-caseating epithelioid cell granulomas found on tissue biopsy. Diagnosis of sarcoidosis requires exclusion of other causes of granuloma formation. Sarcoidosis is also characterized by distinctive laboratory abnormalities, including hyperglobulinemia, an elevated serum angiotensin converting enzyme level, evidence of depressed cellular immunity manifested by cutaneous anergy and, occasionally, hypercalcemia and hypercalciuria. Glucocorticoids remain the mainstay of therapy when treatment is required, although other anti-inflammatory agents are being used increasingly often. PMID- 9861879 TI - Practical management of treatment-resistant depression. AB - Patients receiving antidepressant monotherapy may be partially or totally resistant to treatment in 10 to 30 percent of cases. In patients who have experienced only partial treatment results, the clinician should first consider optimizing antidepressant dosage or lengthening therapy. Antidepressant drug substitution should generally be reserved for use in patients who haven't responded at all (nonresponders). Combining two or more antidepressants is generally not recommended, as this approach may obscure adequate monotherapy evaluation and lead to significant adverse effects or drug-drug interactions. Use of electroconvulsive therapy is recommended in patients with psychotic and severe refractory depression. Augmentation therapy is often efficacious in patients who exhibit a partial antidepressant response. Lithium and thyroid hormone have been the most extensively studied augmentative agents but, more recently, pindolol and buspirone have also been used for this purpose. PMID- 9861880 TI - Senile dementia of the Binswanger's type. AB - Senile dementia of the Binswanger's type is a term used to describe a dementia syndrome characterized by onset in the sixth or seventh decade of life, subcortical neurologic deficits, psychiatric disorders and evidence of hypertension or systemic vascular disease. The status of senile dementia of the Binswanger's type as a distinct entity is a matter of some controversy. The array of neuroimaging abnormalities and clinical findings attributed to this condition overlap with a number of other neuropathologies. Leukoaraiosis, or attenuation of subcortical white matter, seen on computed tomographic scans or magnetic resonance imaging of the brain, is a hallmark of senile dementia of the Binswanger's type. The clinical findings associated with Binswanger's disease are varied but typically include a progressive dementia, depression and "subcortical" dysfunction such as gait abnormalities, rigidity and neurogenic bladder. Treatment is largely supportive and includes a discussion about advanced directives, social support and antidepressant therapy. Control of hypertension and aspirin prophylaxis may help prevent further progression of white matter disease. PMID- 9861881 TI - Appropriate use of the intrauterine device. AB - The intrauterine device, a common form of birth control in the early 1970s, is now avoided by American physicians and women because of concern about complications. This concern is largely the result of the problems reported with use of an intrauterine device that is no longer manufactured. More recent intrauterine devices have an improved design, and reevaluation has shown them to be a safe, efficacious and cost-effective form of birth control. Careful patient selection and preinsertion counseling are crucial to success with the device. Recent studies conclude that the intrauterine device poses no increased risk of pelvic inflammatory disease or infertility when used by appropriately selected patients. PMID- 9861882 TI - Photo quiz. Chronic cough. PMID- 9861883 TI - An AIDS patient's right to refuse life-sustaining treatment. PMID- 9861885 TI - Significant decrease in AIDS deaths in 1997. PMID- 9861884 TI - U.S. Public Health Service updates guidelines for HIV prophylaxis in health care workers. PMID- 9861886 TI - Once-daily drug for persons with HIV. PMID- 9861887 TI - [The 13C-methacetin breath test for quantitative noninvasive liver function analysis with an isotope-specific nondispersive infrared spectrometer in liver cirrhosis]. AB - BACKGROUND AND OBJECTIVE: Cytochrome-P450-dependent liver function can be measured with the 13C-methacetin breath test (MBT). This has heretofore been done with the use of a mass-spectrometer. This study was undertaken to evaluate the MBT (NDIRS) done with the isotope-selective nondispersive infrared spectrometer. PATIENTS AND METHODS: 20 healthy volunteers (ten women, ten men, aged 22-76 years) and 16 patients (ten women, six men, aged 48-71 years) with histologically confirmed liver cirrhosis (Child-Pugh stage A [n = 7], B [n = 5] or C [n = 4]) were given 13C-methacetin in 100 ml of tea after a 12-hour fasting period. Breath tests were performed before the test drink and 5, 10, 15, 20, 30, 40, 50, 60, 80, 100, 120, 150 and 180 min thereafter. The ratio of 13C to 12C was determined, as delta (/1000), and from it the maximal percentage rate (PDRmax) calculated, as well as the cumulative rate (cPDRmax) after 30, 60, 120 and 180 min (Median and 5th and 95th percentiles). RESULTS: For patients with liver cirrhosis there were significantly lower values for cPDRmax and cPDR after 30, 60, 120 and 180 min than in the healthy subjects (P < 0.002): PDR [%]/h: 3.9 (0.7-15.9) vs. 36.5 (23.1-50.0); cDPR 30 min [%]: 1.1 (-0.2-6.0) vs. 12.4 (7.6-17.1); cDPR 3 h [%]: 9.8 (-2.3-27.5) vs. 36.0 (29.9-45.1). There were significant differences among the patients, depending on their Child-Pugh staging. CONCLUSION: The MBT with the cost-effective NDIRS can reliably and noninvasively distinguish between healthy subjects and patients with liver cirrhosis. The test is therefore suitable for the quantitative analysis of liver functions. PMID- 9861888 TI - [The coexistence of 2 different neuroendocrine tumors of the upper gastrointestinal tract and pancreas]. AB - HISTORY AND CLINICAL FINDINGS: A 41-year-old obese patient presented with cramp like abdominal pain, watery diarrhoea with partly digested food particles, projectile vomiting and newly diagnosed diabetes mellitus. For the preceding 6 years he had been treated for recurrent gastric and duodenal ulcers. Although the fasting gastrin level was raised and Zollinger-Ellison syndrome suspected, computed tomography (CT), magnetic resonance imaging (MRI) and coeliac angiography at another hospital had failed to discover a tumor. INVESTIGATIONS: Biochemical tests were unremarkable except for an increased GPT concentration, slight fasting hyperglycemia and hypertriglyceridemia. The gastrin and chromogranin A levels were markedly elevated (15,590 pg/ml and 584.2 U/l, respectively). Gastroscopy revealed, in addition to multiple small duodenal ulcers, a round polypoid mass (diameter of 0.7 cm) lateral to the papilla of Vater, histologically an APUDoma. Endoscopic retrograde cholangiopancreatography (ERCP) revealed a 0.5 cm long compression of the duct of Wirsung in the region of the head of the pancreas. Liver metastases were excluded by magnetic resonance imaging and computed tomography. Endosonography showed a ca. 4 mm space-occupying lesion in the region of the body of the pancreas. Octreotide scintigraphy demarcated two foci at the level of the head of the pancreas (somatostatin receptor positive). TREATMENT AND COURSE: After a pylorus-preserving partial duodenopancreatectomy with lymph node dissection N1/N2, histology confirmed a gastrinoma of the duodenum and a glucagonoma of the pancreas (pT3pN1pMx). Postoperatively the patient became symptom-free and both the blood sugar level and the tumor marker were normal. CONCLUSION: Combined ERCP, endosonography and scintigraphy are more sensitive than other radiological examinations (CT and MRI) in diagnosing and localizing neuroendocrine tumours of the gastrointestinal tract. Despite the low incidence of such tumours, the possible synchronous occurrence of several such tumour should not be ignored. PMID- 9861890 TI - [Hepatic hydrothorax. Recommendations for a rational therapy]. PMID- 9861889 TI - [Manganese superoxide dismutase-inhibiting autoantibodies in cholestatic Epstein Barr viral hepatitis]. AB - HISTORY AND CLINICAL FINDINGS: A 21-year-old woman reported no serious previous illness. For 3 days before admission she had a fever, headache and joint pains. She had become progressively more jaundiced. Physical examination was normal except for enlarged liver and spleen, swollen lymph nodes and facial oedema. INVESTIGATIONS: GOT (30 U/l), GPT (33 U/l) and alkaline phosphatase (172 U/l) were slightly elevated. Serum bilirubin was raised to 12.4 mg/dl. The total white blood cell count was normal, but there were 45% atypical lymphocytes (activated T lymphocytes). Abdominal sonography and endoscopic retrograde cholangiopancreatography were unremarkable. Serology for hepatitis A, B and C as well as for antimitochondrial antibodies was negative, but there were specific IgM (1:640) and IgG antibodies (1:80) against Epstein-Barr virus (EBV) capsid antigen in the immunofluorescence test. DIAGNOSIS, TREATMENT AND COURSE: The EBV infection (infectious mononucleosis) was complicated by cholestatic hepatitis. High concentrations (1832 Gottingen units/ml) of enzyme-inhibiting autoantibodies against the antioxidative enzyme manganese-superoxide dismutase (MSD) were demonstrated. The autoantibodies reduced the antioxidative action of the enzyme by more than 70% and favoured the oxidative cell damage in vitro. After bed-rest for one week without further treatment the symptoms improved and the abnormal laboratory values, including the autoantibodies against MSD, regressed. CONCLUSION: Autoantibodies against MSD are formed during an acute infection with EBV. Their enzyme-inhibiting action promotes abnormalities of oxidative cell function and may thus be the cause of cholestatic hepatitis in this infection. PMID- 9861891 TI - [Alcohol and myocardial infarct. Epidemiological and experimental studies on the effect of alcohol on vascular relaxation and coronary sclerosis]. PMID- 9861892 TI - [The duty of fee reduction according to Article 6 a GOA (Gebuhrenordnung fur Arzte) and expenses reimbursement in the furnishing of inpatient services by established physicians. The judgement of the Federal High Court of 14 January 1998]. PMID- 9861893 TI - [Cachexia due to Sjogren's syndrome]. PMID- 9861894 TI - When healing medicines harm. PMID- 9861895 TI - Geriatrics photo quiz. Amyloidosis. PMID- 9861896 TI - Annular lesions with beaded borders. PMID- 9861897 TI - Late-life psychosis: making the diagnosis and controlling symptoms. AB - Psychosis is best considered as a state of brain dysfunction characterized by delusions, hallucinations, and formal thought disorder. The greatest risk factor for the development of late-life psychosis is the presence of a progressive dementia. Management of the psychotic patient incorporates nonpharmacologic and pharmacologic interventions. Neuroleptics are the most common treatment for psychosis; they generally have very similar efficacy, and thus the choice of agent generally depends on the side effect profile. The newer atypical neuroleptics have relatively fewer anticholinergic and extrapyramidal effects and therefore are often good choices for the older population. PMID- 9861898 TI - Immunologic bullous diseases: blisters without a precipitating event. AB - Although the overall incidence of immunologic bullous diseases is comparatively low, they are nonetheless potentially lethal dermatologic disorders that occur most commonly in individuals age 55 and older. Thus familiarity with the signs, symptoms, and treatments will be valuable to the primary care physician. For bullous diseases in general, the challenge is to differentiate between those arising from immunologic rather than exogenous causes, such as drug-induced or drug-triggered pemphigus. Treatment goals include screening for associated malignancies, managing the lesions, and minimizing the morbidity and mortality associated with the disease. Collaboration with a dermatologist can aid in achieving these objectives. PMID- 9861899 TI - Clubbing: a tipoff to underlying disease. PMID- 9861900 TI - The use of antidepressants in long-term care and the geriatric patient: geriatric psychiatry issues. PMID- 9861901 TI - The use of antidepressants in long-term care and the geriatric patient: primary care issues. PMID- 9861902 TI - Antidepressant pharmacodynamics, pharmacokinetics, and drug interactions. PMID- 9861903 TI - How much should we be spending on health services for Aboriginal and Torres Strait Islander people? PMID- 9861905 TI - United Kingdom Prospective Diabetes Study: the end of the beginning? PMID- 9861904 TI - Should research ethics change at the border? PMID- 9861906 TI - A national approach to skin cancer prevention: the National SunSmart Schools Program. PMID- 9861907 TI - The management of elderly patients with femoral fractures. A randomised controlled trial of early intervention versus standard care. AB - OBJECTIVE: To determine the effect of an early intervention program in an acute care setting on the length of stay in hospital of elderly patients with proximal femoral fractures. SETTING: Acute orthopaedic ward of a large teaching hospital. DESIGN AND PARTICIPANTS: A randomised controlled trial comparing 38 Intervention patients with 33 Standard Care patients. INTERVENTION: Early surgery, minimal narcotic analgesia, intense daily therapy and close monitoring of patient needs via a multidisciplinary approach versus routine hospital management. MAIN OUTCOME MEASURES: Length of stay (LOS); deaths; level of independent functioning. RESULTS: Mean LOS was shorter in the Intervention group than in the Standard Care group (21 days v. 32.5 days; P < 0.01). After adjusting for other factors that could affect LOS (eg, age, sex, pre-trauma functional levels, pre-trauma comorbidity and postsurgical complications), the Intervention program was significantly predictive of shorter LOS (P = 0.01). The Intervention group did not experience greater numbers of deaths, deterioration in function or need for social support than the Standard Care group. CONCLUSION: This early intervention program in an acute care setting results in significantly shorter length of hospital stay for elderly patients with femoral fractures. PMID- 9861908 TI - Home-based treatment of cellulitis with twice-daily cefazolin. AB - OBJECTIVE: To assess the clinical outcome and pharmacokinetics of therapy with cefazolin for patients with cellulitis in a hospital-in-the-home (HIH) program. DESIGN: Observational study with outcome data compared with previously published reports of therapy for cellulitis. SETTING: A university teaching hospital and HIH unit, July 1996-December 1997. PARTICIPANTS: Patients with cellulitis were eligible for inclusion provided their medical condition was stable, they did not require surgical intervention, and their social circumstances allowed home-based therapy. INTERVENTION: Cefazolin 2 g intravenously twice daily, with regular nursing and medical assessment. MAIN OUTCOME MEASURES: Clinical efficacy; peak and trough serum concentrations of cefazolin. RESULTS: Fifty-seven patients (37 were men) with a mean age of 48 years (range, 18-90 years) had 61 episodes of moderate to severe cellulitis (41, lower limb; 17, upper limb; and three, face). They received a median of 11 doses of cefazolin (range, 3-27 doses). Clinical outcomes were: cure in 54, improvement in one, treatment failure in three, and in the remaining three episodes the outcome was indeterminate. Cefazolin concentrations were measured in 27 patients. All peak concentrations were more than 40 micrograms/mL, while trough concentrations were all above the MIC90 of the expected pathogens: median, 3.2 micrograms/mL (range: 0.4-18.5 micrograms/mL). Cefazolin was well tolerated. CONCLUSIONS: Twice-daily cefazolin 2 g intravenously is a convenient and effective option for home-based treatment of patients with cellulitis. Its clinical efficacy is comparable with other treatment regimens. PMID- 9861909 TI - Serotonin syndrome resulting from drug interactions. AB - We describe six patients diagnosed with serotonin syndrome after exposure to drugs with serotonergic activity. Drug interactions occurred as a result of a combination of tricyclic antidepressants, selective serotonin reuptake inhibitors, selective noradrenaline reuptake inhibitors or monoamine oxidase inhibitors. Management included supportive care and the use of non-specific serotonin antagonists (cyproheptadine, benzodiazepines and chlorpromazine). All patients made uneventful recoveries. PMID- 9861910 TI - Reducing indigenous mortality in Australia: lessons from other countries. AB - Mortality rates from all causes in Maoris in New Zealand and Native Americans have fallen substantially since the early 1970s. Comparable mortality rates for Australian Aboriginals and Torres Strait Islanders in 1990-1994 were at or above the rates observed 20 years ago in Maoris and Native Americans, being 1.9 times the rate in Maoris, 2.4 times the rate in Native Americans, and 3.2 times the rate for all Australians. Circulatory diseases, respiratory diseases, injuries and endocrine diseases (mostly diabetes) are responsible for almost 70% of these excess deaths. Mortality rate trends in indigenous populations in other countries suggest the feasibility of substantial and rapid reductions in mortality rates of Australia's indigenous people. PMID- 9861911 TI - Heparin in acute ischaemic stroke. The T wave is negative and it's time to stop. AB - The use of heparin for acute ischaemic stroke has long been controversial. Although it may have some theoretical benefits in preventing fibrin formation and thrombus propagation, data from 16 randomised controlled trials involving more than 22,000 patients show that heparin produces no significant net benefit in eventual outcome. PMID- 9861912 TI - Polycystic ovary syndrome: a new direction in treatment. AB - Polycystic ovary syndrome is a diagnosis made in 5%-10% of women between late adolescence and the menopause. Patients may present with oligomenorrhoea or amenorrhoea, anovulation or infertility, hirsutism or acne. Women with the syndrome have at least seven times the risk of myocardial infarction and ischaemic heart disease of other women, and by the age of 40 years up to 40% will have type 2 diabetes or impaired glucose tolerance. Polycystic ovary syndrome is associated with insulin resistance, with consequent hyperinsulinaemia and (frequently) hyperlipidaemia and obesity. Recent research has shown that the application of diabetes management techniques aimed at reducing insulin resistance and hyperinsulinaemia (such as weight reduction and the administration of oral hypoglycaemic agents) can not only reverse testosterone and luteinising hormone abnormalities and infertility, but can also improve glucose, insulin and lipid profiles. The management of polycystic ovary syndrome should now include patient education and attention to diabetes and cardiovascular risk factors such as hyperlipidaemia, obesity, physical exercise, glucose intolerance, hypertension and cigarette smoking. PMID- 9861913 TI - Clinical trials in developing countries: scientific and ethical issues. AB - Since the 1994 finding that intensive zidovudine treatment of mothers and infants can dramatically reduce perinatal transmission of human immunodeficiency virus, this treatment has been widely adopted in developed countries. In developing countries, trials of less-intensive (and cheaper) regimens have gone ahead, many funded by foreign governments and the United Nations. Controversy has erupted over these trials, particularly over their use of placebo controls. Do differences in healthcare needs and budgets justify different ethical standards in the developed and the developing world? PMID- 9861914 TI - Gastro-oesophageal reflux disease. AB - Gastro-oesophageal reflux disease is the most common cause of indigestion in the community, and is usually chronic. Typical symptoms are recurrent retrosternal burning (heartburn) and regurgitation of sour or bitter fluid. In patients with typical symptoms and no alarm symptoms (pain on swallowing, dysphagia, weight loss or anaemia), treatment may be instituted without investigation. Patients with alarm symptoms and those who respond poorly or relapse after initial treatment require investigation (endoscopy and possibly pH monitoring). About 60% of reflux sufferers have no evidence of mucosal injury; their management aims to relieve symptoms. About 40% of reflux sufferers have oesophagitis and/or complications such as Barrett's oesophagus or oesophageal stricture at endoscopy. Drug therapy consists of H2-receptor antagonists, cisapride or proton-pump inhibitors. PMID- 9861915 TI - Toddler drownings in domestic swimming pools in Queensland since uniform fencing requirements. PMID- 9861916 TI - Peritonsillar abscess. PMID- 9861917 TI - Prevalence of MRSA in South Australian nursing homes. PMID- 9861918 TI - Budesonide substitution in Crohn's disease relieves CNS toxicity of systemic steroids. PMID- 9861919 TI - Should folate be added to flour to prevent neural tube defects? PMID- 9861920 TI - Trends in general practice in the Waikato, 1979-80/1991-92, II: Social variations in service use and clinical activity. AB - AIMS: To document trends in ethnic group and occupational class differences in the use of general practitioner services and in patterns of clinical activity for the Hamilton Health District of the Waikato over the period 1979-80 to 1991-92. METHODS: The data are drawn from a baseline and a follow-up survey of general practice in the Waikato region representing a one per cent sample of all in surgery, in-hours, week-day encounters at two points in time. The data were recorded by participating general practitioners in four collection weeks spaced over the period of a year. In total, 9468 and 10,235 patient encounter forms were completed respectively. RESULTS: Over a period in which service availability and rates of medical contact grew, there was a relatively greater increase in utilisation among Maori and lower socioeconomic groups: between the two surveys the ratio of Maori to non-Maori rates increased from 0.8 to 1.0 and the ratio of visits for lower to higher socioeconomic groups grew from a differential of 2.5 to one of 3.1. More serious conditions apart, these changes seemed to occur uniformly regardless of the severity, amenability or susceptibility of the condition presented to the general practitioner. Changes in ethnic group and occupational class patterns of service activity almost exactly mirrored these trends. CONCLUSIONS: A notable relative increase in rates of contact for primary medical care among Maori and lower socioeconomic groups seems to have accompanied the growth in the 1980s of the availability of general practitioner services in this region of New Zealand. PMID- 9861921 TI - When not to syringe an ear. AB - This article reviews the potential hazards of ear syringing and the Accident Compensation Corporation Medical Misadventure Unit's experience of alleged iatrogenic injury over a 17-month period. Syringing claims account for about 25% of the total claims received by the ACC ENT Medical Misadventure Committee. Forty seven claims were accepted as either mishap or error, and 63 claims were declined. Perforation of the drum was by far the commonest injury resulting in significant disability. Severe iatrogenic otitis externa accounted for most of the remainder. Practice nurses had syringed the ears of approximately two-thirds of the claimants with significant disability. The contraindications to ear syringing are listed and discussed. The correct technique, with a few "do and don't" pointers, is briefly reviewed. Management of the unintentional injury is discussed. PMID- 9861922 TI - Chronic solvent neurotoxicity in New Zealand: notified cases between 1993 and 1997. AB - AIM: To report on cases of chronic solvent neurotoxicity notified to the Department of Labour between 1993 and 1997. METHODS: Previously published diagnostic criteria were used to classify notified cases as "verified", "not verified" or "not proven". Verified cases were further classified as Type 1 or 2(2) and analysed according to occupation, solvent type and length of exposure. RESULTS: Of 193 notified cases, 76 were classified as "verified". The majority were male (74/76) and European (69/76). Mean length of exposure to solvents for Type I cases was 10.6 years (SD 4.7) and for Type II cases 19.8 years (SD 8.4). Forty-three cases were classified as "mild" and 33 as "moderate". The most frequent occupation was spraypainting (39%), followed by printing (16%) and boatbuilding (9%). There was no correlation between severity of symptoms and type of solvent. There was a non-significant trend of increasing severity of symptoms with length of exposure. CONCLUSION: Exposure to solvents is an important health hazard in New Zealand workplaces. PMID- 9861923 TI - Ocular toxicity from ethambutol: a review of four cases and recommended precautions. AB - AIMS: To document the clinical and demographic features of cases of ethambutol ocular toxicity, to review the literature on this subject and to critically review current guidelines for ethambutol administration. METHODS: Cases of ocular toxicity from ethambutol were sought retrospectively at Green Lane and Wellington Hospitals between 1992 and 1995. The records of cases identified were examined. RESULTS: Four subjects with tuberculosis developed ocular toxicity 2 1/2, 7 1/2, 8 and 12 months after starting ethambutol. Normal visual acuity returned in three cases; one patient has severe, permanent visual impairment. Language difficulties were present in three subjects. CONCLUSIONS: Impaired communication was potentially very important in this series. Special care is needed in educating patients about ethambutol. We propose additional recommendations: 1. the usual daily dose of ethambutol should be 15 mg/kg/day, not 25 mg/kg/day; using 25 mg/kg/day (or lesser doses in the presence of renal impairment) should prompt regular formal ophthalmological evaluation (e.g. monthly) in cases with comprehension or communication difficulties; 3. both ethambutol and isoniazid should be stopped immediately if severe optic neuritis occurs. Isoniazid should be stopped if less severe optic neuritis does not improve within six weeks after stopping ethambutol. PMID- 9861924 TI - An audit of magnetic resonance imaging in the paediatric orthopaedic setting. AB - AIMS: To determine when magnetic resonance imaging (MRI) is helpful in the management of a selected group of paediatric orthopaedic patients. METHODS: A retrospective analysis of 131 MRI scans was undertaken with allocation into seven categories based on clinical presentation. RESULTS: MRIs performed for spinal, congenital and intra-articular pathology, as well as for growth plate assessment correlated well with subsequent clinical and/or surgical findings. Three of ten MRIs (30%) incorrectly assessed whether a foreign body was present with a sensitivity of 0.60 and specificity of 0.80. Three of 20 MRIs (15%) could not accurately distinguish between oedema/effusion and frank infection. Here sensitivity was 1.00 and specificity was 0.73. MRIs performed for assessment of tumours were accurate with respect to margins and extent. Assessment of pathology with MRI was never the indication for MRI, thus it was not surprising that in four of 27 soft tissue tumours (sensitivity of 0.88 and specificity of 0.63) MRI did not correlate with subsequent histological findings. CONCLUSIONS: MRI plays an important role in the assessment of a wide range of musculo-skeletal pathology. MRI does not, and could not be expected to, replace the need for incisional biopsy for tumour diagnosis. It must be used with caution where tissues have been previously explored for foreign bodies. Interpretation of MRI in musculoskeletal infection must consider its timing in the evolution of the patient's infection. PMID- 9861925 TI - Second response to immunoglobulin in recurrent Guillain-Barre syndrome. AB - Guillain-Barre, syndrome is the most common paralytic illness among healthy adults. With modern critical care, the mortality has fallen although prolonged hospitalisation and significant morbidity are common. Plasma exchange and intravenous immunoglobulin have only recently been shown to be equally efficacious; the combination of plasma exchange plus intravenous immunoglobulin does not confer additional advantage. Contrary to earlier fears, immunoglobulin use is not associated with an increased relapsed rate. Some patients do benefit from a second course of immunoglobulin in recurrent Guillain-Barre syndrome. PMID- 9861926 TI - Neonatology: will the infant learn to speak? PMID- 9861927 TI - Dr Eliza Foster McDonogh Frikart: advertiser. AB - Eliza Frikart was the first woman doctor registered in New Zealand. She moved round Australia and New Zealand and advertised her presence wherever she went. She was struck off the British register. PMID- 9861928 TI - PHARMAC and statins. PMID- 9861929 TI - New Zealand death certificates. PMID- 9861930 TI - Handlebar accidents. PMID- 9861931 TI - New birth and death registration forms. PMID- 9861932 TI - Patient privilege. PMID- 9861933 TI - Sex; death in agony: AIDS reversed. PMID- 9861934 TI - Hyperbaric oxygen therapy for children with cerebral palsy. PMID- 9861935 TI - Nosocomial transmission of tuberculosis to health care workers in Mpumalanga. PMID- 9861936 TI - The South African Allergic Rhinitis Working Group and allergic rhinitis. PMID- 9861937 TI - Organ electrodermal diagnostics. PMID- 9861938 TI - First aid treatment for burns in children--a call for education. PMID- 9861939 TI - The humanities in medicine and the open food challenge. PMID- 9861940 TI - Viagra--it's no free ride. PMID- 9861941 TI - Asthma in South Africa--a long way to go! PMID- 9861943 TI - To all doctors--an open letter. PMID- 9861942 TI - New warnings on the use of isotretinoin (Roaccutane) PMID- 9861944 TI - Mad cows revisited. PMID- 9861947 TI - Problem-based learning in clinical clerkship--the experience at the University of Transkei Medical School. PMID- 9861945 TI - Challenges in health science education. PMID- 9861946 TI - Noxious toads and frogs of South Africa. AB - The major defence mechanism in frogs in via the secretion of toxins from their skin. In humans, intoxication may occur when part of the amphibian integument is ingested, as in the form of herbal medicines. Two groups of South African frogs have skin secretions that are potentially lethal to humans and animals. Toads (Bufo and Schismaderma species), the amphibians with which man and his pets most frequently have contact, secrete potent toxins with cardiac glycoside activity. Topical and systemic intoxication, while seen in humans, remains predominantly a veterinary problem. Intoxication by the red-banded rubber frog, which secretes an unidentified cardiotoxin, is far less common. The probable mechanisms of intoxication and management of a poisoned patient are discussed. PMID- 9861948 TI - Osteoporosis in clinical practice--bone densitometry and fracture risk. AB - Osteoporosis is a condition of decreased bone mass and bone density associated with an increase in fracture risk. Bone mineral density (BMD) of the lumbar spine and femur can be reliably measured by double-beam X-ray absorptiometry (DEXA), which provides a measure of bone strength. Reduction in BMD is a continuum and is associated with a progressive increase in fracture risk. The diagnosis of osteoporosis is based on BMD relative to that of healthy young adults and criteria for diagnosis are arbitrary. The original 'normal' BMD data published by some manufacturers were relatively high, leading to a relative over-diagnosis of osteoporosis. Revised normative BMD values of the spine and femur and revised criteria using degrees of severity are proposed and may provide a better basis for diagnosis and for the management of patients with osteoporosis. The indications for BMD measurement, the age at which BMD is measured, and number of measurements, depends upon the purpose of the measurement and how the result will affect the management of each patient in clinical practice. PMID- 9861949 TI - An assessment of the 'road-to-health' card based on perceptions of clinic staff and mothers. AB - OBJECTIVES: To describe the opinions of health personnel and parents at child health clinics in Cape Town; to determine the accuracy and completeness of data recorded on the present 'Road-to-Health' (RTH) card; and to ascertain the views of clinic staff and mothers regarding what information they would like to record. DESIGN: Descriptive prospective study. SETTING AND SUBJECTS: Qualitative interviews of 35 health personnel and 150 mothers/caregivers were conducted at 17 child health clinics. The clinic practices of 32 health personnel were monitored and details of 150 RTH cards were examined. MAIN OUTCOME MEASURES: Responses of health personnel at public and private child health clinics and of mothers were analysed. Data recorded on the card were extracted under the headings: neonatal data, immunisation schedules, measurements, and weight-for-age chart. RESULTS: Most nurses supported the concept of an RTH card but a large majority recommended that it be replaced with a notebook retained by the mother. A significant proportion of health personnel did not know how to use the weight-for-age chart. Most mothers attending clinics carried the card, but this number dropped for hospital visits and consultations with private doctors. Mothers' understanding of the card was limited. For mothers the weight-for-age chart, immunisation schedule and milestone section are obscure. CONCLUSION: Health personnel and mothers would like to replace the RTH card with a notebook in the parents' home language. It should contain more information on health matters, adequate space to record weight and infectious diseases, an illustrated milestone chart and an improved schedule for immunisations. Mothers perceived the RTH card as belonging to the clinic and wanted a 'baby's own' document. PMID- 9861951 TI - Residents in endemic malaria areas of South Africa are complacent about the dangers of malaria. PMID- 9861950 TI - Successful plague control in Namibia. AB - OBJECTIVE: To demonstrate that plague can be successfully controlled. DESIGN: A descriptive study outlining patterns of plague occurrence in relation to variables such as age group, gender, place and time. SETTING: Two northern districts, namely Engela in Ohangwena region and Onandjokwe in Oshikoto region, an area of 2,000 km2. SUBJECTS: All patients who presented to the health facilities with signs and symptoms of plague were considered. Diagnosis was made on the basis of clinical symptomatology and laboratory confirmation. OUTCOME MEASURES: A plague control programme was established involving the following components: management capability at the local level, case recognition and management, dusting programme, rodent trapping programme, health education, establishment of plague laboratory, and plague surveillance system. RESULTS: Following the establishment of the control programme plague cases were reduced from 1,092 to zero within 3 years and deaths from 45 to zero within 2 years. The case fatality rate was reduced from 4.12% to 0% over a 3-year period. No cases have been reported in Namibia for the past 3 years. CONCLUSION: The Namibian experience has demonstrated that plague can be controlled through a combination of strategies taking local conditions into consideration. PMID- 9861952 TI - Severe encephalopathies in children with antibodies reactive with Rickettsia africae. PMID- 9861953 TI - Potential risk of enhancing survival of infected cells. PMID- 9861954 TI - Survival time in cats questioned. PMID- 9861955 TI - Dr. Abraham Lincoln Eisenhower--pioneer veterinarian and uncle of a president. PMID- 9861956 TI - Reshaping veterinary medical education: the British experience. PMID- 9861957 TI - What is your diagnosis? Hepatic mass involving the left liver lobes. PMID- 9861958 TI - Rabies surveillance in the United States during 1997. AB - In 1997, 49 states, the District of Columbia, and Puerto Rico reported 8,509 cases of rabies in nonhuman animals and 4 cases in human beings to the Centers for Disease Control and Prevention. Nearly 93% (7,899) were wild animals, whereas 7% (610) were domestic species. The total number of reported cases increased 19.4% from that of 1996 (7,128 cases). Increases were apparent in each of the major species groups, with the exception of cattle. The relative contributions of these groups to the total reported for 1997 were as follows: raccoons (50.5%; 4,300 cases), skunks (24.0%; 2,040), bats (11.3%; 958), foxes (5.3%; 448), cats (3.5%; 300), dogs (1.5%; 126), and cattle (1.4%; 122). The 958 cases of rabies reported in bats represented a 29.3% increase over the total reported for 1996 and the greatest number reported since 1984, with cases reported by 46 of the 48 contiguous states. The epizootic of rabies in raccoons expanded into Ohio in 1997 and now includes 19 states and the District of Columbia. Thirteen states, where rabies in raccoons is enzootic, reported increases over 1996 in total numbers of reported cases. New York (1,264 cases), North Carolina (879), Virginia (690), and Maryland (619) reported the greatest numbers of cases [corrected]. Five states reported increases that exceeded 50%, compared with cases reported in 1996: Ohio (673.3%; 15 cases in 1996 to 116 in 1997). Massachusetts (144.3%; 115 to 281), South Carolina (97.9%; 96 to 190), Connecticut (97.4%; 274 to 541), and Maine (86.3%; 131 to 244). Cases of rabies associated with foci of rabies in foxes in west central Texas and in dogs and coyotes in southern Texas continued to decline, with this state reporting 78.3% fewer rabid foxes (13 cases), 26.7% fewer rabid dogs (11), and 63.2% fewer rabid coyotes (7) during 1997, compared with 1996. Reported cases of rabies in cats (300) and dogs (126) increased 12.8% and 13.5%, respectively, whereas cases in cattle (122) decreased by 6.9%. Thirty states, the District of Columbia, and Puerto Rico reported increases in rabies in animals during 1997, compared with decreases reported by 31 states and the District of Columbia in 1996. One state (Mississippi; 5 cases) remained unchanged. Hawaii was the only state that did not report a case of rabies in 1997. Four indigenously acquired cases of rabies reported in human beings were the result of infection with rabies virus variants associated with bats. PMID- 9861959 TI - Salmonella enteritidis infections in the United States. PMID- 9861960 TI - Salmonella Typhimurium DT104 in cattle in Great Britain. PMID- 9861961 TI - Escherichia coli O157:H7 outbreaks in the United States, 1982-1996. PMID- 9861962 TI - Campylobacter jejuni in foods. PMID- 9861963 TI - The issues of residues and human health. PMID- 9861964 TI - The president's national food safety initiative. PMID- 9861965 TI - Impact of antimicrobic use in veterinary medicine. PMID- 9861966 TI - Hazard Analysis and Critical Control Point systems as a preventive tool. PMID- 9861967 TI - Use of competitive exclusion in food animals. PMID- 9861968 TI - Microbial risk assessment, economics, and food safety. PMID- 9861969 TI - Synopsis of the postharvest food safety symposium. PMID- 9861970 TI - Cytologic examination of exfoliative specimens obtained during endoscopy for diagnosis of gastrointestinal tract disease in dogs and cats. AB - OBJECTIVE: To determine whether cytologic examination of exfoliative specimens obtained during endoscopy was as useful as histologic examination of mucosal biopsy specimens for the diagnosis of gastrointestinal tract disease in dogs and cats and to compare the diagnostic accuracy of 2 techniques (brush or touch) in preparing specimens for cytologic examination. DESIGN: Prospective case series. ANIMALS: 85 dogs and 23 cats. PROCEDURE: Specimens for cytologic and histologic examination were obtained during routine endoscopic examination of the stomach, small intestine, and colon. A diagnosis was made on the basis of cytologic findings (graded objectively) and compared with the diagnosis on the basis of histologic findings. RESULTS: The diagnostic accuracy of cytologic examination was high for all 3 organs. Sensitivities, specificities, and predictive values of positive and negative results were > 90% in most instances. The diagnostic accuracy of the brush technique was equal or superior to that of the touch technique for 84% of specimens. The brush technique was most useful in detecting cellular infiltrates in the lamina propria, whereas the touch technique was more likely to detect acute mucosal inflammation. Percentages of false-positive (3.2%) and false-negative (6.9%) cytologic interpretations were low. CLINICAL IMPLICATIONS: Endoscopy is safe and requires little time to procure specimens for cytologic examination, which can be obtained concurrently with mucosal biopsy specimens. Cytologic examination of exfoliative specimens obtained during endoscopy is a useful and reliable adjunct to histologic examination of biopsy specimens in the diagnosis of gastrointestinal tract disease in dogs and cats. PMID- 9861971 TI - Efficacy of clomipramine in the treatment of canine compulsive disorder. AB - OBJECTIVE: To assess the efficacy of clomipramine for treatment of canine compulsive disorder (CCD). DESIGN: Randomized, placebo-controlled, double-blind, balanced AB-BA crossover clinical study. ANIMALS: 51 dogs with CCD. PROCEDURES: Dogs were given clomipramine (3 mg/kg [1.3 mg/lb] of body weight, PO, q 12 h) for 4 weeks and placebo for 4 weeks. At the end of each treatment each owner rated the severity of their dog's behavior, using 2 validated rating scales. Statistical analysis was made by ordinal regression. Compliance, adverse effects, and the effectiveness of masking were also assessed. Each dog's behavior was reevaluated 1 to 2 years after completing the study. RESULTS: Behaviors included spinning (n = 17) and self-mutilation by licking (acral lick dermatitis, 12). Both rating scales demonstrated a treatment effect. Compliance was satisfactory, and masking was effective. Sedation and reduced appetite were reported more commonly when dogs were given clomipramine than when they were given placebo. Forty-five dogs available for follow-up evaluation still had their behaviors; 6 dogs were lost to follow-up evaluation. CLINICAL IMPLICATIONS: Results suggest that clomipramine was effective in dogs with CCD and was not associated with serious adverse effects. However, treatment for 4 weeks was not curative. Behavior modification is likely to be necessary to manage CCD. PMID- 9861972 TI - Detection and effects of helicobacters in healthy dogs and dogs with signs of gastritis. AB - OBJECTIVES: To determine prevalence, colonization density, and distribution of helicobacters and gastric histologic findings in healthy dogs and dogs with signs of gastritis; to evaluate association of colonization density and gastric inflammation; and to compare the number of Helicobacter spp with degree of inflammation. DESIGN: Cross-sectional prevalence survey. ANIMALS: 25 healthy dogs and 21 dogs with signs of gastritis. PROCEDURE: During endoscopy, gastric mucosal biopsy specimens were obtained from healthy and affected client-owned dogs. Histologic and cytologic evaluation and results of a urease test were used for detecting helicobacters, which were identified definitively by use of transmission electron microscopy and bacterial culture. RESULTS: Helicobacters were detected in all 25 healthy and 20 of 21 affected dogs. Cytologic examination was a more sensitive method than histologic examination or the urease test. Helicobacters were found least frequently and in fewest number in the antrum in both groups of dogs. Gastric inflammation was evident in both groups of dogs and did not differ significantly between groups. A significant association was not detected between colonization density or the number of Helicobacter spp and degree of gastric inflammation. In both groups, H bizzozeronii, H felis, and H salomonis were cultured. CLINICAL IMPLICATIONS: Histologically verified chronic gastritis is common in dogs with signs of gastritis as well as in healthy dogs. Colonization density of helicobacters was not associated with degree of gastric inflammation in the dogs of our study. It remains to be determined whether certain strains of Helicobacter spp can induce gastritis in dogs. PMID- 9861974 TI - Surgical removal of fragmented coronoid processes and fractured anconeal process in an older dog with evidence of severe degenerative joint disease. AB - A 10-year-old Labrador Retriever was admitted because of severe unilateral (left) forelimb lameness of 6 weeks' duration. Computerized tomography revealed bilateral fragmented coronoid processes (FCP) and unilateral fracture of the anconeal process. Surgery on the left elbow to remove the loose anconeal process and FCP resolved the severe lameness and improved the dog's overall activity, compared with that of the preceding 2 years. Unstable FCP can develop late in life, and a degenerative anconeal process may fracture. Surgical removal of loose fragments in a severely arthritic joint may be beneficial. PMID- 9861973 TI - Possible antibiotic-associated colitis in a dog. AB - A Poodle referred for renal disease developed severe colonic disease characterized by total mucosal collapse and necrosis. The onset of colonic disease was temporally related to administration of antibiotics. On 3 occasions, bacterial culture of fecal samples yielded only Streptococcus pyogenes, Pseudomonas aeruginosa, and Candida albicans, suggesting that the colonic bacterial flora was severely disrupted. Findings, although not conclusive, were suggestive of antibiotic-associated colitis that ultimately proved fatal. Colonoscopy should be considered for dogs with unduly severe large-bowel diarrhea associated with antibiotic treatment and can be done with minimal restraint and bowel preparation, if necessary. Nonselective bacterial culture of fecal samples should be considered for dogs with unduly severe large-bowel diarrhea associated with antibiotic treatment. PMID- 9861975 TI - Abdominal lipomatosis attributed to tall fescue toxicosis in deer. AB - Five adult female Eld's deer died acutely or were euthanatized because of clinical signs including anorexia, signs of depression, and uremia. On necropsy, these deer had large masses of necrotic abdominal fat constricting the ureters, causing hydroureter and hydronephrosis. The herd from which these deer originated was maintained on pastures consisting primarily of tall fescue, samples from which were subsequently confirmed to be infected with an endophytic fungus that is known to cause similar lesions in cattle. A retrospective study of deaths in this herd revealed a sharp increase in incidence of abdominal lipomatosis since 1994. Physical examinations on the herd revealed > 90% of females to be affected. Endophyte-infected tall fescue forage was concluded to be a major factor in the development of lipomatosis in these deer. Other contributing factors were considered. Lesions caused by endophyte-infected fescue can be severe, and this disease can develop in nondomestic species. PMID- 9861976 TI - The design of matching pursuit filters. AB - This paper presents a new technique for creating efficient and compact models from data, called matching pursuit filters. The design of a matching pursuit filter is based on an adapted wavelet expansion, where the expansion is adapted to both the data and the pattern recognition problem being addressed. This contrasts with most adaptation schemes, where the representation is a function of the data, but not of the problem to be solved. This approach does not decompose the images in the training set individually, but rather determines the expansion by simultaneously decomposing all the images. Because it uses two-dimensional wavelets as the building blocks for the decomposition, the representation is explicitly two-dimensional and is composed of local information. Matching pursuit filters can be trained to detect, recognize, or identify objects and have been applied to recognizing faces and detecting objects in infrared imagery. PMID- 9861977 TI - Local interaction fields and adaptive regularizers for surface reconstruction and image relaxation. AB - A set of novel local interaction fields associated with input point data and adaptive regularizers are introduced for some problems in image processing and early-middle vision. The local interaction field is used to take the place of the delta-error term in conventional approaches. It consists of two parts; one is the usual error term, the other is a window function whose shape, size and orientation can be adapted to local structures in specific applications. Unlike the usual delta-error term, local interaction fields favour local flatness and thus impose smoothing implicitly. Several adaptive regularizers are introduced to keep discontinuities while smoothing. In finding solutions, even the simplest gradient descent algorithm is efficient in many examples. The results are stable under varying parameters and quite robust against noise. PMID- 9861978 TI - Theory and implementation of infomax filters for the retina. AB - In the first part of this paper we discuss a technical visual sensory system, which--in analogy with the retina--includes some preprocessing of visual information. In so doing, we use an information-theoretic criterion, the infomax ansatz, to optimize the response of the sensory system. In particular, it is shown that the lattice structure of the photoreceptor array has to be taken into account. By a discrete Fourier transformation on a triangular lattice we derive the frequency response of the infomax filter within the first Brillouin zone. To illustrate the response properties, infomax filters adapted to different noise levels are applied to images with different signal-to-noise ratios. This clearly demonstrates the necessity of adaptation of the filter properties to the given noise level. Furthermore, it is shown how to efficiently implement infomax-like filters by simple networks with only nearest-neighbour interactions. A two layered network topology proves to be very advantageous in implementing the desired high-pass or low-pass properties. The network topology allows for adaption of the network to low and high noise levels by simply adjusting the nearest-neighbour couplings. In the second part of this paper, we compare the previously described information-theoretic requirements on a visual sensory system with biological facts known from the vertebrate retina. The substantial physiological response properties of the vertebrate retina are in agreement with the main features of the infomax filter. Since available experimental data lacks information which is necessary for a more quantitative comparison, we present suggestions for future experiments. Some key anatomical features of the retina of many vertebrates compare well with our two-layered implementation of the infomax filter. The analogy, in particular, concerns the adaption mechanism. To illustrate this point, we summarize some recent experiments which demonstrate that in the retina of some species adaption is based on the release of the neuromodulator dopamine by the interplexiform cells. This causes the horizontal cells to decouple. On the basis of recently gained understanding of the outer plexiform layer of the retina some further hypotheses about the functionality of the retina become obvious and possible future experiments to verify or refute them are suggested. Finally, we discuss the infomax approach from a more general point of view. In particular, we show that redundancy is essential to obtaining noise robustness of an internal representation of the environment as it is produced by a sensory system such as the retina. PMID- 9861979 TI - Cascaded redundancy reduction. AB - We describe a method for incrementally constructing a hierarchical generative model of an ensemble of binary data vectors. The model is composed of stochastic, binary, logistic units. Hidden units are added to the model one at a time with the goal of minimizing the information required to describe the data vectors using the model. In addition to the top-down generative weights that define the model, there are bottom-up recognition weights that determine the binary states of the hidden units given a data vector. Even though the stochastic generative model can produce each data vector in many ways, the recognition model is forced to pick just one of these ways. The recognition model therefore underestimates the ability of the generative model to predict the data, but this underestimation greatly simplifies the process of searching for the generative and recognition weights of a new hidden unit. PMID- 9861980 TI - Contextual quick-learning and generalization by humans and machines. AB - In a previous study (1994 Network: Comput. Neural Syst. 5 203-27) we compared human quick-learning and generalization (quick modelling) with that of neural nets (feedforward architectures), symbolic algorithms (decision tree procedures), and pattern classifiers (truth-set descriptors). Those studies raised the question of the role of context in the nature and rapidity of human learning. Here we address that issue in the setting of the same basic experiment (Quinlan classification problem) used for the previous studies. A major implication of our findings is that humans overwhelmingly seek, create, or imagine context in order to provide meaning when presented with abstract or apparently incomplete or contradictory or otherwise untenable situations. PMID- 9861981 TI - Pattern matching in a model of dendritic spines. AB - Pattern matching, the ability to recognize and maximally respond to an input pattern that is similar to a previously learned pattern, is an essential step in any learning process. To investigate the properties of pattern matching in biological neurons, and in particular the role of a calcium-dependent potassium conductance, a circuit model of a small area of dendritic membrane with a number of dendritic spines is developed. Circuit model simulations show that dendritic membrane depolarization is greater in response to a previously learned pattern of synaptic inputs than in response to a novel pattern of synaptic inputs. These simulations, in combination with an analysis of the circuit model equations, reveal that when a synaptic input pattern is similar to the learned pattern of synaptic inputs, the total dendritic depolarization is a linear combination of dendritic depolarization contributed by individual spines. When at least one synaptic input differs markedly from the learned value, dendritic depolarization is a nonlinear combination of individual spine depolarizations. These principles of spine interactions are captured in a computationally simple set of 'similarity measure' equations which are shown to reproduce the response surface of the circuit model output. Thus, these similarity measure equations not only describe a biologically plausible model of pattern matching, they also satisfy computational requirements for use in artificial neural networks. PMID- 9861982 TI - Slow stochastic Hebbian learning of classes of stimuli in a recurrent neural network. AB - We study unsupervised Hebbian learning in a recurrent network in which synapses have a finite number of stable states. Stimuli received by the network are drawn at random at each presentation from a set of classes. Each class is defined as a cluster in stimulus space, centred on the class prototype. The presentation protocol is chosen to mimic the protocols of visual memory experiments in which a set of stimuli is presented repeatedly in a random way. The statistics of the input stream may be stationary, or changing. Each stimulus induces, in a stochastic way, transitions between stable synaptic states. Learning dynamics is studied analytically in the slow learning limit, in which a given stimulus has to be presented many times before it is memorized, i.e. before synaptic modifications enable a pattern of activity correlated with the stimulus to become an attractor of the recurrent network. We show that in this limit the synaptic matrix becomes more correlated with the class prototypes than with any of the instances of the class. We also show that the number of classes that can be learned increases sharply when the coding level decreases, and determine the speeds of learning and forgetting of classes in the case of changes in the statistics of the input stream. PMID- 9861983 TI - Recall of old and recent information. AB - A neural network model for 'forgetting upon learning' is developed in such a way that old information can also be evoked. A new synaptic clipping scheme coupled with selective reinforcement of information is introduced that mimics the learning and memory functions of the 'limbic system' in the brain. The model thus enables both long-term (old) and short-term (recent) memories to exist concurrently, without one affecting the other, as one expects in a realistic situation. PMID- 9861984 TI - Modelling multiple-cause structure using rectification constraints. AB - We present an artificial neural network which self-organizes in an unsupervised manner to form a sparse distributed representation of the underlying causes in data sets. This coding is achieved by introducing several rectification constraints to a PCA network, based on our prior beliefs about the data. Through experimentation we investigate the relative performance of these rectifications on the weights and/or outputs of the network. We find that use of an exponential function on the output to the network is most reliable in discovering all the causes in data sets even when the input data are strongly corrupted by random noise. Preprocessing our inputs to achieve unit variance on each is very effective in helping us to discover all underlying causes when the power on each cause is variable. Our resulting network methodologies are straightforward yet extremely robust over many trials. PMID- 9861985 TI - Learning attractors in an asynchronous, stochastic electronic neural network. AB - LANN27 is an electronic device implementing in discrete electronics a fully connected (full feedback) network of 27 neurons and 351 plastic synapses with stochastic Hebbian learning. Both neurons and synapses are dynamic elements, with two time constants--fast for neurons and slow for synapses. Learning, synaptic dynamics, is analogue and is driven in a Hebbian way by neural activities. Long term memorization takes place on a discrete set of synaptic efficacies and is effected in a stochastic manner. The intense feedback between the nonlinear neural elements, via the learned synaptic structure, creates in an organic way a set of attractors for the collective retrieval dynamics of the neural system, akin to Hebbian learned reverberations. The resulting structure of the attractors is a record of the large-scale statistics in the uncontrolled, incoming flow of stimuli. As the statistics in the stimulus flow changes significantly, the attractors slowly follow it and the network behaves as a palimpsest--old is gradually replaced by new. Moreover, the slow learning creates attractors which render the network a prototype extractor: entire clouds of stimuli, noisy versions of a prototype, used in training, all retrieve the attractor corresponding to the prototype upon retrieval. Here we describe the process of studying the collective dynamics of the network, before, during and following learning, which is rendered complex by the richness of the possible stimulus streams and the large dimensionality of the space of states of the network. We propose sampling techniques and modes of representation for the outcome. PMID- 9861986 TI - Nonlinear feedforward networks with stochastic outputs: infomax implies redundancy reduction. AB - We prove that maximization of mutual information between the output and the input of a feedforward neural network leads to full redundancy reduction under the following sufficient conditions: (i) the input signal is a (possibly nonlinear) invertible mixture of independent components; (ii) there is no input noise; (iii) the activity of each output neuron is a (possibly) stochastic variable with a probability distribution depending on the stimulus through a deterministic function of the inputs (where both the probability distributions and the functions can be different from neuron to neuron); (iv) optimization of the mutual information is performed over all these deterministic functions. This result extends that obtained by Nadal and Parga (1994) who considered the case of deterministic outputs. PMID- 9861987 TI - Development of localized oriented receptive fields by learning a translation invariant code for natural images. AB - Neurons in the mammalian primary visual cortex are known to possess spatially localized, oriented receptive fields. It has previously been suggested that these distinctive properties may reflect an efficient image encoding strategy based on maximizing the sparseness of the distribution of output neuronal activities or alternately, extracting the independent components of natural image ensembles. Here, we show that a strategy for transformation-invariant coding of images based on a first-order Taylor series expansion of an image also causes localized, oriented receptive fields to be learned from natural image inputs. These receptive fields, which approximate localized first-order differential operators at various orientations, allow a pair of cooperating neural networks, one estimating object identity ('what') and the other estimating object transformations ('where'), to simultaneously recognize an object and estimate its pose by jointly maximizing the a posteriori probability of generating the observed visual data. We provide experimental results demonstrating the ability of such networks to factor retinal stimuli into object-centred features and object-invariant transformation estimates. PMID- 9861988 TI - A model of cortical associative memory based on a horizontal network of connected columns. AB - An attractor network model of cortical associative memory functions has been constructed and simulated. By replacing the single cell as the functional unit by multiple cells in cortical columns connected by long-range fibers, the model is improved in terms of correspondence with cortical connectivity. The connectivity is improved, since the original dense and symmetric connectivity of a standard recurrent network becomes sparse and asymmetric at the cell-to-cell level. Our simulations show that this kind of network, with model neurons of the Hodgkin Huxley type arranged in columns, can operate as an associative memory in much the same way as previous models having simpler connectivity. The network shows attractor-like behaviour and performs the standard assembly operations despite differences in the dynamics introduced by the more detailed cell model and network structure. Furthermore, the model has become sufficiently detailed to allow evaluation against electrophysiological and anatomical observations. For instance, cell activities comply with experimental findings and reaction times are within biological and psychological ranges. By introducing a scaling model we demonstrate that a network approaching experimentally reported neuron numbers and synaptic distributions also could work like the model studied here. PMID- 9861989 TI - Spatio-temporal influences at the neural level of object recognition. AB - In late 1988, Miyashita published work reporting recordings of single cells in the inferotemporal cortex of the macaque monkey (Miyashita 1988 Nature 335 817 20). He described the responses of neurons to a sequence of random fractal pattern images, and how many of the neurons tested were seen to respond strongly to a subset of the images on the basis of sequence presentation order, i.e. appearance in time, rather than their spatial similarity. In this work, I describe a local Hebb-like learning rule which in conjunction with a simple feedforward neural architecture is capable of replicating the type of temporal order association apparent in the cells from which he made recordings. The paper also advances reasons for requiring such learning by describing its possible role in establishing transformation invariant representations of objects. PMID- 9861990 TI - Generalization and exclusive allocation of credit in unsupervised category learning. AB - A new way of measuring generalization in unsupervised learning is presented. The measure is based on an exclusive allocation, or credit assignment, criterion. In a classifier that satisfies the criterion, input patterns are parsed so that the credit for each input feature is assigned exclusively to one of multiple, possibly overlapping, output categories. Such a classifier achieves context sensitive, global representations of pattern data. Two additional constraints, sequence masking and uncertainty multiplexing, are described; these can be used to refine the measure of generalization. The generalization performance of EXIN networks, winner-take-all competitive learning networks, linear decorrelator networks, and Nigrin's SONNET-2 network are compared. PMID- 9861991 TI - On the matter of rules. Past-tense formation and its significance for cognitive neuroscience. AB - If neuronal models are successful, they will account for specifically human complex behaviours. Most of these behaviours can be described as governed by rules. In recent years, much effort has been spent to elucidate the neuronal basis of rules, and many researchers have focussed on modelling rules and regularities underlying language, particularly those relevant for past-tense formation. After introducing problems posed by past-tense formation, important aspects of recent controversies between connectionists and linguists concerning the nature of rules will be reviewed and analysed on the basis of elementary simulations. It is argued that modular networks with varying connection probabilities between their layers would be ideal for modelling learning and processing of past-tense formation. The motivation for postulating such networks comes from neurobiological models of language and from neuroanatomical data about cortico-cortical connectivity. Furthermore, such modular networks may explain double dissociations of regular and irregular past-tense formation in neurological patients, as reported in recent neuropsychological publications. It is concluded that past-tense formation does not pose problems to pattern associators, given that some structure is built into the network which approximates wirings in the human cortex. PMID- 9861992 TI - The coding of information by spiking neurons: an analytical study. AB - We analyse analytically the coding of information by a spiking neuron. The emphasis is on the question of how many spikes are necessary for the reliable discrimination of two different input signals. The discrimination ability is measured by the second-order Renyi mutual information between the random variable describing the name of the signal and a sequence of n output spikes. Analysing this measure as a function of n, we study the coding strategy of a single spiking neuron, with the following main results. A small number of output spikes is required for efficient discrimination of input signals, i.e. for encoding them, if the separation is easy; a large number of output spikes is required in the difficult case of separation of very similar input signals. Three different versions of the spike response model of a single neuron are studied. The approach presented can be regarded as a non-parametric version of the reconstruction method of Bialek. PMID- 9861993 TI - Spatial and temporal pattern analysis via spiking neurons. AB - Spiking neurons, receiving temporally encoded inputs, can compute radial basis functions (RBFs) by storing the relevant information in their delays. In this paper we show how these delays can be learned using exclusively locally available information (basically the time difference between the pre- and postsynaptic spikes). Our approach gives rise to a biologically plausible algorithm for finding clusters in a high-dimensional input space with networks of spiking neurons, even if the environment is changing dynamically. Furthermore, we show that our learning mechanism makes it possible that such RBF neurons can perform some kind of feature extraction where they recognize that only certain input coordinates carry relevant information. Finally we demonstrate that this model allows the recognition of temporal sequences even if they are distorted in various ways. PMID- 9861994 TI - Irregular synchronous activity in stochastically-coupled networks of integrate and-fire neurons. AB - We investigate the spatial and temporal aspects of firing patterns in a network of integrate-and-fire neurons arranged in a one-dimensional ring topology. The coupling is stochastic and shaped like a Mexican hat with local excitation and lateral inhibition. With perfect precision in the couplings, the attractors of activity in the network occur at every position in the ring. Inhomogeneities in the coupling break the translational invariance of localized attractors and lead to synchronization within highly active as well as weakly active clusters. The interspike interval variability is high, consistent with recent observations of spike time distributions in visual cortex. The robustness of our results is demonstrated with more realistic simulations on a network of McGregor neurons which model conductance changes and after-hyperpolarization potassium currents. PMID- 9861995 TI - Autonomous development of decorrelation filters in neural networks with recurrent inhibition. AB - We perform a quantitative analysis of information processing in a simple neural network model with recurrent inhibition. We postulate that both excitatory and inhibitory synapses continually adapt according to the following Hebbian-type rules: for excitatory synapses correlated pre- and post-synaptic activity induces enhanced excitation; for inhibitory synapses it induces enhanced inhibition. Following synaptic equilibration in unsupervised learning processes, the model is found to perform a novel type of principal-component analysis which involves filtering and decorrelation. In the light of these results we discuss the possible role of the granule-/Golgi-cell subnetwork in the cerebellum. PMID- 9861996 TI - Learning algorithms based on linearization. AB - The aim of this article is to investigate a mechanical description of learning. A framework for local and simple learning algorithms based on interpreting a neural network as a set of configuration constraints is proposed. For any architectural design and learning task, unsupervised and supervised algorithms can be derived, optionally using unconstrained and hidden neurons. Unlike algorithms based on the gradient in weight space, the proposed tangential correlation (TC) algorithms move along the gradient in state space. This results in optimal scaling properties and simple expressions for the weight updates. The number of synapses is much larger than the number of neurons. A constraint for neural states does not impose a unique constraint for synaptic weights. Which weights to assign credit to can be selected from a parametrization of all weight changes equivalently satisfying the state constraints. At the heart of the parametrization are minimal weight changes. Two supervised algorithms (differing by their parametrizations) operating on a three-layer perceptron are compared with standard backpropagation. The successful training of fixed points of recurrent networks is demonstrated. The unsupervised learning of oscillations with variable frequencies is performed on standard and more sophisticated recurrent networks. The results presented here can be useful both for the analysis and for the synthesis of learning algorithms. PMID- 9861997 TI - A simple model for neural computation with firing rates and firing correlations. AB - A simple extension of standard neural network models is introduced which provides a model for neural computations that involve both firing rates and firing correlations. Such an extension appears to be useful since it has been shown that firing correlations play a significant computational role in many biological neural systems. Standard neural network models are only suitable for describing neural computations in terms of firing rates. The resulting extended neural network models are still relatively simple, so that their computational power can be analysed theoretically. We prove rigorous separation results, which show that the use of firing correlations in addition to firing rates can drastically increase the computational power of a neural network. Furthermore, one of our separation results also throws new light on a question that involves just standard neural network models: we prove that the gap between the computational power of high-order and first-order neural nets is substantially larger than shown previously. PMID- 9861998 TI - Learning viewpoint-invariant face representations from visual experience in an attractor network. AB - In natural visual experience, different views of an object or face tend to appear in close temporal proximity as an animal manipulates the object or navigates around it, or as a face changes expression or pose. A set of simulations is presented which demonstrate how viewpoint-invariant representations of faces can be developed from visual experience by capturing the temporal relationships among the input patterns. The simulations explored the interaction of temporal smoothing of activity signals with Hebbian learning in both a feedforward layer and a second, recurrent layer of a network. The feedforward connections were trained by competitive Hebbian learning with temporal smoothing of the post synaptic unit activities. The recurrent layer was a generalization of a Hopfield network with a low-pass temporal filter on all unit activities. The combination of basic Hebbian learning with temporal smoothing of unit activities produced an attractor network learning rule that associated temporally proximal input patterns into basins of attraction. These two mechanisms were demonstrated in a model that took grey-level images of faces as input. Following training on image sequences of faces as they changed pose, multiple views of a given face fell into the same basin of attraction, and the system acquired representations of faces that were approximately viewpoint-invariant. PMID- 9861999 TI - The influence of neural activity and intracortical connectivity on the periodicity of ocular dominance stripes. AB - Several factors may interact to determine the periodicity of ocular dominance stripes in cat and monkey visual cortex. Previous theoretical work has suggested roles for the width of cortical interactions and the strength of between-eye correlations. Here, a model based on an explicit optimization is presented that allows a thorough characterization of how these and other parameters of the afferent input could affect ocular dominance stripe periodicity. The principle conclusions are that increasing the width of within-eye correlations leads to wider columns, and, surprisingly, that increasing the width of cortical interactions can sometimes lead to narrower columns. PMID- 9862000 TI - Experimentally induced pulmonary arterial occlusion with detachable balloon in pigs: thin-section CT findings. AB - RATIONALE AND OBJECTIVES: The authors evaluated changes of lung attenuation in pigs, with special attention to the mosaic pattern of low attenuation, at thin section computed tomography (CT) after obstruction of the proximal pulmonary artery with a detachable balloon. MATERIALS AND METHODS: In seven pigs, nine sites of the descending pulmonary artery were obstructed with detachable balloons. This-section CT scans of the lungs were obtained immediately (n = 9) and at 1 week (n = 5), 2 weeks (n = 1), 3 weeks (n = 2), 4 weeks (n = 1), 6 weeks (n = 1), 8 weeks (n = 1), and 12 weeks (n = 1) after pulmonary artery obstruction. RESULTS: No statistically significant difference was found between the measured lung attenuation of the normal lung and that of the lung distal to the obstruction. Of the nine sites of pulmonary artery obstruction, five (56%) showed an irregular area of increased lung attenuation without lobular architecture. The diameter of the pulmonary artery after obstruction, compared with the diameter before obstruction, decreased by a range of 13%-57% (mean, 35%) and by 0-67% (mean, 44%) at levels 1 cm and 2 cm distal to the obstruction, respectively. CONCLUSION: This experimental study reveals that regional low attenuation areas do not develop for up to 12 weeks after the obstruction of proximal pulmonary artery, despite a marked decrease in the diameter of the pulmonary artery distal to the obstruction. PMID- 9862001 TI - Long-term outcomes of balloon dilation of esophageal strictures in children. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine the long term success of the use of angioplasty balloons for dilation of esophageal strictures in children. MATERIALS AND METHODS: The authors studied 24 children who had undergone fluoroscopically guided angioplasty balloon dilation of esophageal strictures. Ten children had stricture of the anastomosis after surgical repair of esophageal atresia; five had achalasia; three had strictures due to gastroesophageal reflux; three had corrosive strictures; two had congenital stenosis; and one had stricture after radiation therapy. Success was defined as resolution of the dysphagia a year after dilation. RESULTS: A year after the last dilation, 15 of the 24 children had no dysphagia. Patients in whom treatment was successful included eight children with strictures after surgery for esophageal atresia, one with achalasia, three with strictures caused by gastroesophageal reflux, one with a corrosive stricture, one with congenital stenosis, and one with a stricture resulting from radiation therapy. CONCLUSION: The long-term results of balloon catheter dilation are highly successful in patients with stricture after surgical repair of esophageal atresia and stricture due to gastroesophageal reflux. Dilation is not as valuable in the treatment of esophageal strictures resulting from other causes. PMID- 9862002 TI - Second-generation three-dimensional reconstruction for rotational three dimensional angiography. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to assess the feasibility and accuracy of three-dimensional (3D) reconstruction techniques for digital subtraction angiography (DSA) in planning and evaluation of minimally invasive image-controlled therapy. MATERIALS AND METHODS: Using a standard, commercially available system, the authors acquired DSA images and corrected them for inherent distortions. They designed and implemented parallel and multiresolution versions of cone-beam reconstruction techniques to reconstruct high-resolution targeted volumes in a short period of time. Testing was performed on anatomically correct, calibrated in vitro models of a cerebral aneurysm. These models were used with a pulsatile circulation circuit to allow for blood flow simulation during DSA, computed tomographic (CT) angiography, and magnetic resonance (MR) angiography image acquisitions. RESULTS: The multiresolution DSA-based reconstruction protocol and its implementation allowed the authors to achieve reconstruction times and levels of accuracy for the volume measurement of the aneurysmal cavities that were considered compatible with actual clinical practice. Comparison with data obtained from other imaging modalities shows that, besides vascular tree depiction, the DSA-based true 3D technique provides volume estimates at least as good as those obtained from CT and MR angiography. CONCLUSION: The authors demonstrated the feasibility and potential of true 3D reconstruction for angiographic imaging with DSA. On the basis of the model testing, this work addresses both the timing and quantification required to support minimally invasive image-controlled therapy. PMID- 9862003 TI - Radiologic investigations and pathologic results of experimental chronic pancreatitis in cats. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to evaluate a variety of methods to induce chronic pancreatitis and its radiologic expression by experimentally inducing this condition in cats. MATERIALS AND METHODS: Chronic inflammatory and fibrosing pancreatitis was produced in cats by intraductal injection of 1.5 mL of 94% ethanol in one group or by a combination of intraductal and intraparenchymal injection of ethanol together with partial obstruction of the main pancreatic duct to 70% of its original lumen by fixation of a small catheter in the papilla. For comparison, other cats underwent total obstruction of the main pancreatic duct. All groups, as well as untreated control cats (n = 3), underwent repeat laparotomy to obtain biopsy specimens. RESULTS: Cats with total obstruction showed progressing fibrosis with dilatation of ductules occasionally infiltrated with granulocytes. From 26 weeks on, acini and islets of Lnagerhans became atrophic. Radiographs showed progressive but diffuse dilatation of ducts, ductules, and side branches. Cats from the other two groups had interlobular inflammation and fibrosis with flattened and irregular ductular epithelium. Later, ductular proliferation occurred, interstitial inflammation subsided, and fibrosis increased. Radiographs showed very irregular ducts and ductules with stenosis and dilatation. From 26 weeks on, no substantial differences were observed between the cats who received only intraductal injection of ethanol and the cats who underwent the combination of procedures. CONCLUSION: The histopathologic and radiographic alterations that evolved from damage to the ductal epithelium in the cat resembled the features of chronic pancreatitis in humans and differed from those caused by total obstruction of the main pancreatic duct in cats. PMID- 9862004 TI - Electron-beam CT in the noninvasive assessment of coronary stent patency. AB - RATIONALE AND OBJECTIVES: Coronary artery stents reduce the rate of restenosis in patients who have undergone balloon angioplasty; therefore, the implantation of coronary stents represents an important method in the treatment of coronary stenoses. The authors' purpose was to investigate the usefulness of electron-beam computed tomography (CT) as a noninvasive means of assessing the patency of coronary artery stents in patients who had undergone balloon angioplasty and stent placement. MATERIALS AND METHODS: Electron-beam CT was used to assess stent patency in 177 patients with 285 stents. Contrast material-enhanced multisection flow studies were performed, and the images were evaluated by three investigators and compared with the findings of coronary angiography. RESULTS: Cine loop evaluations and time-attenuation curve analysis led to the correct diagnosis in 167 (94.3%) patients, as confirmed with coronary angiography. Stenoses had occurred in 18 of the 194 vessels with stents, and 14 of these were detected with electron-beam CT. CONCLUSION: Electron-beam CT appears to be a valuable imaging modality in the noninvasive assessment of stent patency in coronary arteries. PMID- 9862005 TI - Electron-beam CT angiography with three-dimensional reconstruction in the evaluation of coronary artery bypass grafts. AB - RATIONALE AND OBJECTIVES: The authors evaluated the use of electron-beam computed tomographic (CT) angiography and three-dimensional (3D) reconstruction for evaluation of coronary artery bypass grafts (CABGs). MATERIALS AND METHODS: Sixty patients (53 men and seven women; mean age, 56 years +/- 8) with 152 CABGs (29 internal mammary artery and 123 saphenous vein grafts) underwent electron-beam CT with 3D reconstruction. The time from bypass surgery to CT scanning was 7 days to 120 months (mean, 17 months +/- 28). Enhanced single-section and flow mode studies were performed in all cases. The results were correlated with CABG operation records, and patency was determined with time-attenuation curves. RESULTS: All patients were successfully examined. On the basis of time attenuation curve criteria, 124 (81.6%) of 152 grafts were patent. The patency rate for the internal mammary artery subgroup was 93% (27 of 29 grafts); for the saphenous vein graft subgroup, 78.9% (97 of 123 grafts). CONCLUSION: Used in contrast-enhanced, single-section mode and flow studies with time-attenuation curves, electron-beam CT angiography with 3D reconstruction can provide quantitative data for evaluation of CABG patency. PMID- 9862006 TI - Perinatal morbidity and mortality rates in twin pregnancies--a 15-year review study from Athens. AB - There were 150 deliveries of twin pregnancies over 26 weeks in our hospital. The incidence of twin pregnancies was 1.0% in a population consisting of Greek nationals. Seventy five (50%) were term at delivery whereas 75 (50%) were premature (mean gestational age at delivery 36.5 weeks). The birth weight at delivery ranged from 350 grs to 4,050 grs with mean birth weight for the 1st and 2nd twin of 2,404 grs and 2,384 grs, respectively. One hundred and forty four (48%) neonates had weights between 1,501 to 2,500 grs. Twenty two (9.57%) neonates had weights between 1,501 grs to 2,500 grs. The perinatal mortality when birth weights exceeded 1,500 grs was 7.32%. In total, 186 (62%) newborns were delivered vaginally and 114 (38%) by cesarean section. Forceps were used in 4.33% of the cases and breech extraction was performed in 2.33%. The mode of delivery and perinatal mortality rates were not correlated for both twins (p > 0.05). The most common causes of death were hyaline membrane disease (48.5%), immaturity (18.2%), congenital abnormalities (15.1%), septicemia (12.1%) and asphyxia (6.06%). Prematurity seems to constitute the most common cause of perinatal deaths while the mode of delivery found to be irrelevant to perinatal mortality. PMID- 9862007 TI - Absence of pericentromeric heterochromatin (9qh-) in a patient with bilateral retinoblastoma. AB - The polymorphisms of constitutive heterochromatin regions, present on chromosomes 1, 9, 16 and Y, are inherited in a Mendelian fashion. The C-band heteromorphism has been reported to be associated with various types of cancer. Heterochromatin is considered to play a role in protecting genome against the mutagens. Changes in the quantity and proportion of the different types of satellite DNA might increase the genetic susceptibility in people with heterochromatic variations, which in turn cause chromosome instability and predispose the individual to cancer. We report a case of bilateral retinoblastoma with complete absence of pericentromeric heterochromatin on one of the chromosomes number 9. A similar deficiency of pericentromeric heterochromatin on chromosome number 9 and 16 has been reported in a phenotypically normal individual and a Down syndrome case, respectively. This deficiency was found to be inherited from the father in all the three cases. Complete absence of pericentromeric heterochromatin of chromosome 9 is not being reported in association with cancer syndromes. Further studies are necessary to understand the role of this factor in normals and in those with cancer susceptibility, specially with retinoblastoma and the paternal origin of this deficiency. PMID- 9862008 TI - Ultrasound examination in twin pregnancy and late fetal death. AB - OBJECTIVE: To test the hypothesis that the (case) twin later to succumb in utero has biparietal diameter measurements (by ultrasound) different from those (control) twins surviving the perinatal period. MATERIAL AND METHODS: Information from the Medical Birth Registry, National Board of Health and Welfare, Stockholm, was used to identify all births in a defined population in southern Sweden with about 20,000 deliveries each year. In 3,019 twin pregnancies between 1973 and 1989, one or both twins were stillborn in 47 cases (gestational duration > or = 28 weeks, birthweight > or = 500 g). For each case pregnancy, two control pregnancies were selected, the matching criteria being: same delivery unit, same parity (0, I, II, III+), similar year of delivery (+/- 1 year) and maternal age (+/- 5 years). Data on ultrasound examinations were extracted from the original medical records. Screening in early second trimester started at one of the units as early as 1973 and at the latest of 12 units in 1982. RESULTS: There was no obvious difference between cases and controls in intra-pair discordant biparietal diameter (BPD) measured in early second trimester. Nor was there any evident difference in the rate of deviant BPD between cases and controls. In all, 8% of dead male and 24% of dead female fetuses were by definition small-for-gestational age (< -2 standard deviations). CONCLUSIONS: No significant difference was seen between cases and controls regarding deviating biparietal diameters. Abdominal diameter may be a better predictor of subsequent fetal death (not analysed in this study), though only about 15% of all dead twins were deemed small-for gestational age. PMID- 9862009 TI - Trends of twinning rates in ten countries, 1972-1996. AB - Trends of twinning rates were analyzed using vital statistics in Austria, Finland, Norway, Sweden, Canada, Australia, Hong Kong, Israel, Japan, and Singapore during the period from 1972 to 1996. The twinning rates increased significantly year by year in each country. During these periods, the twinning rate increased by twenty percent in Austria and Canada, and by sixty percent in Norway and Sweden. The twinning rate was 1.6 times higher in Sweden than in Hong Kong in 1972 and by eighty percent in Israel in 1995. Twinning rates were higher in European countries, Canada, and Australia than in Asia. The variations of the twinning rates among countries were not only due to biological factors, but also to assisted reproductive techniques. In Australia, the overall twinning rate was 1.3 times higher in the nuptial births (14.1 per 1000 births) than in the ex nuptial births (10.7) during the period 1994-1996. As for maternal age, twinning rates in Sweden increased year by year for maternal age groups except the youngest and the oldest age groups. In Sweden, the rising twinning rate has been attributed to the higher proportion of mothers (for the 25-39 year old age groups) treated with ovulation-inducing hormones and attributed to in-vitro fertilization. PMID- 9862010 TI - "Mother's child" and "father's child" among twins. A longitudinal twin study from pregnancy to 21 years age, with special reference to development and psychiatric disorders. AB - 234 pairs of twins were studied from pregnancy up to 21 years of age on the basis of records from maternity hospitals, neonatal wards and children's health centres and questionnaires filled in by the parents when the twins were aged 2-10 and 12 21 years, and by the twins themselves at age 12-21. 74 twins were personally interviewed about human relationships in their families and with the Present State Examination (PSE) at age 15-21. When the evaluation of parental preference was made by the parents, the mother's favourites had learned to speak earlier and were more often the psychic leader of the pair, but they more often had sleeping difficulties and other psychosomatic symptoms in adolescence. They were most often scored in class 2-3, non-specific neurotic symptoms in the PSE, but none of them was placed in the higher classes of possible or probable psychiatric disorder. Mothers seem to develop a tighter affectionate bond towards their favourites than do fathers, thus inducing a good basic trust and faster language acquisition in childhood, but probably also transient non-specific neurotic symptoms in adolescence in face of the developmental task of entering autonomous adulthood. The father's favorites were more often the physical leaders of the pair, showed less accident proneness and most often reported tendencies towards autonomy from their co-twins, thus indicating that the fathers' attitudes may be more encouraging towards independence. As the least psychosomatic symptoms were seen in twins in the intermediate position regarding parental preference, it seems reasonable that the division of twins between parents on the grounds of favouritism should not be strict. PMID- 9862011 TI - [A brief history of twin study methods]. PMID- 9862012 TI - Balanced X; 22 translocation in a patient with premature ovarian failure. AB - A case of balanced X; autosome translocation 46, X, t (X; 22) (q 24; q 13) in a 25-year-old female with secondary amenorrhea and premature ovarian failure (POF) is described. The relationship between balanced X; autosome translocation [t (X; A)] and varied phenotypic expression observed in these women is discussed. This case highlights the importance of early recognition of these women in order to give them the best chance of conception in their oligohypomenorrhic phase before complete loss of gonadal function. PMID- 9862013 TI - The acupuncture system and the liquid crystalline collagen fibers of the connective tissues. AB - We propose that the acupuncture system and the DC body field detected by western scientists both in here in the continuum of liquid crystalline collagen fibers that make up the bulk of the connective tissues. Bound water layers on the collagen fibers provide proton conduction pathways for rapid intercommunication throughout the body, enabling the organism to function as a coherent whole. This liquid crystalline continuum mediates hyperreactivity to allergens and the body's responsiveness to different forms of subtle energy medicine. It constitutes a "body consciousness" working in tandem with the "brain consciousness" of the nervous system. We review supporting evidence from biochemistry, cell biology, biophysics and neurophysiology, and suggest experiments to test our hypothesis. PMID- 9862014 TI - Cerebral cortex participation in the physiological mechanisms of acupuncture stimulation: a study by auditory endogenous potentials (P300). AB - Although acupuncture has traditionally used the acupoints formula to treat diseases, the physiological mechanisms involved and the effectiveness of therapy remain unclear. This study investigated the physiological mechanism(s) and response to acupuncture stimulation using the acupoints formula. Scalp-recorded potentials P300 were evoked by auditory stimulation of non-target and target in 13 normal adult volunteers. Latencies and amplitudes were measured. Three assessments were performed in each subject over a period of at least one week. Each assessment was divided into a control period with no acupuncture stimulation, followed by an acupuncture period and then a post-acupuncture period. Acupuncture needles were inserted into the body as follows: 1) non acupoint: acupuncture needles were inserted 2 cm lateral to both Zusanli acupoints; 2) acupoint: acupuncture needles were inserted into both Zusanli acupoints; 3) acupoints formula: acupuncture needles were inserted into both Zusanli and Shousanli acupoints. Our results showed that both acupoint and acupoints formula assessments resulted in a significant decrease of P300 amplitudes during the acupuncture and post-acupuncture periods. However, there was significant difference in P300 amplitudes in the non-acupoint assessment during these periods. P300 changes in latencies and amplitudes were not significantly different between the acupoint assessment and the acupoints formula assessment. We concluded that acupuncture stimulation of both Zusanli acupoints resulted in a decrease of P300 amplitudes, suggesting the involvement of the cerebral cortex in sensory interaction when simultaneous sensations of the two types are received. No similar changes were observed in the non-acupoint assessment, which have been suggested to be related to so-called acupoint specificity. Results obtained using the acupoints formula were not significantly different from those using acupoints alone. These findings suggested that neuropsychological effects from stimulation of Zusanli acupoints and Shousanli acupoints are different. PMID- 9862015 TI - Acute effects of chundosunbup qi-training on blood concentrations of TSH, calcitonin, PTH and thyroid hormones in elderly subjects. AB - The present study investigated how the systemic treatment of a programmed exercise, ChunDoSunBup (CDSB) Qi-training, affects the secretion of thyroid and parathyroid hormones in elderly subjects (10 male and 5 female). Plasma concentrations of thyroid stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), parathyroid hormone (PTH), ionized calcium, and calcitonin were determined. CDSB Qi-training induces a slight increase in TSH. Both T4 and T3 were increased at the mid-time of CDSB Qi-training (p < 0.05). There were significant correlations only between T3 and T4 at mid-training. This shows that increase in the plasma level of T3 was associated with the secretion of T4. The plasma concentrations of calcitonin and PTH were increased at mid-time and post time of CDSB Qi-training. But ionized calcium was decreased slightly by CDSB Qi training. These results suggest that Qi-training modulates the secretion of thyroid hormones, calcium metabolism, and parathyroid hormones in the elderly. However, whether the long-term practice of CDSB Qi-training might change bone metabolism and have longitudinal effects on the thyroid hormone of the elderly need further investigation. PMID- 9862016 TI - The correlation between skin electrical conductance and the score of qi vacuity. AB - In traditional Chinese medicine, the syndrome of qi vacuity means that the patient's body has a low level of energy to react to stress. Recently, we used a score, the QV score, by scaling the severity of symptoms and signs of qi vacuity in patients with tiredness. The aim of this study was to investigate the relationship between QV score and skin electrical conductance in patients with tiredness. One hundred and forty-three healthy controls and 103 patients with tiredness were involved. Each subject received a weak electrical stimulation with constant voltage (1.75 volt), and conductance was measured between two different limbs. The mean value of skin conductance among four limbs was calculated and expressed by a special unit, namely Chin. The correlation between the skin conductance and QV score was analyzed by a linear regression analysis. The results showed that skin electrical conductance of healthy controls was negatively correlated with age (r-coefficient = -0.51, P = 0.000). The skin conductance of patients with tiredness was significantly lower than that of healthy controls with matching age (P = 0.000 by Student's t-test). Moreover, there was a positive correlation between the decrease of skin conductance and the QV score in patients with tiredness (r-coefficient = +0.68, P = 0.000). These results suggest that a decrease in skin electrical conductance may be closely related to the severity of qi vacuity. The skin conductance test is a simple, reliable, and quantitative method for detection of syndrome of qi vacuity. PMID- 9862017 TI - Fructus corni attenuates oxidative stress in macrophages and endothelial cells. AB - The antioxidant effect of a Chinese medicinal herb, Fructus corni extract (FCE), was investigated using models of oxidative stress in macrophages and vascular endothelial cells. Murine macrophages (J774) were incubated with FCE at 37 degrees C and 5% CO2 for 1 hr. Oxidative burst was triggered by zymosan and measured with a fluorescent probe. FCE exhibited a concentration- dependent suppression of oxidative burst. Confluent monolayers of bovine pulmonary artery endothelial cells (PAEC) were preincubated with FCE for 20 hrs, washed, and then exposed to an organic oxidant t-butyl hydroperoxide (tBHP) for 2 hrs. Cell viability was assessed by methylthiazol tetrazolium (MTT) assay, and cell injury by the release of intracellular lactate dehydrogenase (LDH). Lipid peroxidation products of PAEC were determined by measuring thiobarbituric acid-reactive substances (TBARS). Exposure of PAEC to tBHP resulted in decreased cell viability, increased LDH release, and elevated TBARS. Preincubation of PAEC with FCE significantly reversed these changes. Our results demonstrated that FCE can protect vascular endothelial cells from oxidant injury. The data thus suggest that Fructus corni may be useful for the prevention and/or treatment of disorders associated with oxidative damage. PMID- 9862018 TI - The effect of tert-butyl hydroperoxide on peritoneal macrophages and the protective effect of protein-bound polysaccharide administered intraperitoneally and orally. AB - Our previous studies have shown that a protein-bound polysaccharide, polysaccharide krestin (PSK), can protect macrophages from lipoperoxidative injury induced by tert-butyl hydroperoxide (tbOOH) and oxidatively modified low density lipoprotein (O-LDL). PSK was administered intraperitoneally to mice, and the peritoneal macrophages harvested were used as an experimental model. PSK does not reveal any protective effect on macrophages injured by tbOOH when incubated in vitro. In order to elucidate its mode of action, in the present study, the protective effects of PSK were further investigated using morphological changes and viability as indices. It was shown that, compared with the non-PSK group, the viability of macrophages was much higher in the PSK group, and the morphological changes in the PSK group were much less than those in the non-PSK group. The protective effect of PSK administered intraperitoneally and orally on macrophages injured by tbOOH was comparable. Furthermore, the serum of the PSK treated mice had the same protective effect. PMID- 9862019 TI - Analysis of inhibitory effects of scutellariae radix and baicalein on prostaglandin E2 production in rat C6 glioma cells. AB - Inhibitory mechanism of the water extract of Scutellariae Radix on prostaglandin E2 (PGE2) release was examined in C6 rat glioma cells. Scutellariae Radix reduced a Ca2+ ionophore A23187-induced PGE2 release by inhibition of arachidonic acid (AA) liberation. Sho-saiko-to and San'o-shashin-to, which contain Scutellariae Radix, also inhibited PGE2 release. A23187 caused phosphorylation of mitrogen activated protein kinase (MAPK), resulting in activation of cytosolic phospholipase A2 (cPLA2). Scutellariae Radix and baicalein inhibited the phosphorylation of MAPK. Baicalein, but not baicalin, inhibited A23187-induced PGE2 release. These results suggest that baicalein in Scutellariae Radix reduces AA liberation through the inhibition of the MAPK-cPLA2 pathway. PMID- 9862020 TI - The protective effects of traditional Chinese medicine prescription, han-dan-gan le, on CCl4-induced liver fibrosis in rats. AB - Han-Dan-Gan-Le, a Chinese medicine preparation composed of Salvia miltorrhiza, Radix paeoniae, Astragalus membranaceus, Stephania tetrandra, and dried leaves of Ginkgo biloba, has been used successfully to treat human liver fibrosis and cirrhosis for years. This study was designed to examine the mechanisms of the protection. Male Wistar rats were given CCl4 (1.2 ml/kg, 2 times/week), 20% fat diet, and 30% alcohol in drinking water (every other day) for 6 weeks. Han-Dan Gan-Le (0.5 and 1.0 g/kg, p.o., daily for 6 weeks) was administered to rats simultaneously to examine the protective effects against CCl4-induced liver fibrosis. The experimentally-induced liver fibrosis and other morphological alterations were significantly ameliorated by Han-Dan-Gan-Le. Han-Dan-Gan-Le treatments decreased CCl4-induced hepatic collagen accumulation by more than 50%, and significantly increased urinary excretion of hydroxyproline. The CCl4-induced lipid peroxidation in liver and serum was ameliorated as a result of Han-Dan-Gan Le treatment, possibly by restoring the activity of superoxide dismutase activity in liver and erythrocytes, In conclusion, Han-Dan-Gan-Le is effective in protecting against liver fibrosis. The mechanisms of the protection appear to be due to its antioxidant properties and the modulation of hepatic collagen metabolism. PMID- 9862021 TI - Mechanism of the protective effects of sumac gall extract and gallic acid on the progression of CCl4-induced acute liver injury in rats. AB - To examine the mechanism of the preventive effect of tannins on the progression of carbon tetrachloride (CCl4)-induced acute liver injury in rats, sumac gall (SG) extract and gallic acid (GA) were used as substitutes for crude tannins, because SG is a kind of Chinese traditional medicinal herb containing large amounts of various tannins, and GA is one of the major constituents of SG. The protective effect of oral (p.o.) and intraperitoneal (i.p.) administration of each substance on progression of CCl4-induced hepatitis was investigated in rats. Speculating that the superoxide dismutase (SOD)-like activities (O2 radical scavenging activities) and/or protective effects of these substances on cell membranes might play a key role in the mechanism opposing the progression of CCl4 induced hepatitis, the O2 radical-scavenging activities in liver cells and serum in rats were monitored. Both substances significantly prevented the progression of acute liver injury with both p.o. and i.p. administration. These findings suggest that the mechanism for this prevention might be due mainly to the protective effect of these substances on cell membranes rather than O2 radical scavenging activities. PMID- 9862022 TI - Effect of pretreatment of rats with an urinary preparation on liver injuries induced by carbon tetrachloride and alpha-naphthylisothiocyanate. AB - The effect of oral administration of a preparation of human urine (PHU) on the progression of acute liver injury was examined in rats intoxicated with carbon tetrachloride (CCl4) and alpha-naphthylisothiocyanate (ANIT) PHU protected the liver from CCl4-induced injury as judged by morphological and biochemical observations. In contrast, PHU aggravated ANIT-induced injury as judged also by morphological and biochemical observation. PHU prevented the increase in hepatic glutathione (GSH) and lipid peroxidation induced by CCl4. But PHU enhanced the increase in hepatic GSH caused by ANIT. These results indicate that the effect of PHU on hepatic GSH concentrations is through an indirect pathway. Clinical application of PHU on hepatitis should be explored further. PMID- 9862024 TI - The effects of ginseng radix rubra on human vascular endothelial cells. AB - The effect of Ginseng Radix Rubra (Red ginseng) on human vascular endothelial cells was examined. Red ginseng was found to promote the proliferation of vascular endothelial cells, inhibit the production but promote the decomposition of endothelin, which is known to constrict blood vessels and raise blood pressure as well as accelerated the synthesis of nitric oxide, which is known to have an angio-tonic effect. Furthermore, Red ginseng was observed to increase the production of Interleukin 1 beta, which is known to play important roles in the homeostatic activities of the human body such as immunity and inflammation as well as increasing the production of tissue plasminogen activators, which suppress the formation of thrombin in the blood coagulation and fibrinolysis mechanisms. It is suggested that Red ginseng has the effect of accelerating endothelial cells proliferation and of promoting physiological activities. PMID- 9862023 TI - Effects of garlic compounds diallyl sulfide and diallyl disulfide on arylamine N acetyltransferase activity in strains of Helicobacter pylori from peptic ulcer patients. AB - Arylamine N-acctyltransferase (NAT) activities with p-aminobenzoic acid (PABA) and 2-aminofluorene (2-AF) were determined in the bacterium Helicobacter pylori collected from peptic ulcer patients. Two assay systems were performed, one with cellular cytosols, the other with intact cell suspensions. Cytosols or suspensions of H. pylori with or without specific concentrations of diallyl sulfide (DAS) or diallyl disulfide (DADS) co-treatment showed different percentages of 2-AF and PABA acetylation. The data indicated that there was decreased NAT activity associated with increased levels of DAS or DADS in H. pylori cytosols and suspensions. Viability studies on H. pylori demonstrated that DAS or DADS elicited dose-dependent bactericide affects on H. pylori cultures. The data also indicated that DAS and DADS decreased the apparent values of K(m) and Vmax of NAT enzyme from H. pylori in both systems examined. This report is the first demonstration that garlic components can affect H. pylori growth and NAT activity. PMID- 9862025 TI - Effect of Ganoderma lucidum on postherpetic neuralgia. AB - Administration of hot water soluble extracts of Ganoderma lucidum (GI) (36 to 72 g dry weight/day) decreased pain dramatically in two patients with postherpetic neuralgia recalcitrant to standard therapy and two other patients with severe pain due to herpes zoster infection. PMID- 9862026 TI - Sixty-three years with Chinese herbal medicines; memoir of a botanist. PMID- 9862027 TI - [Sex-specific analysis of cartilage volume in the knee joint--a quantitative MRI based study]. AB - The objective of the present study was to determine differences in the normal knee joint cartilage volume of males and females, the analysis of the percentage distribution of the cartilage tissue onto the various joint surfaces, and the determination of the relationship between the cartilage volume, the body weight, and the tibial head diameter. We examined the knee joints of nine healthy men and nine women with a low level of physical activity. The cartilage volume was assessed with magnetic resonance imaging, applying a fat-suppressed gradient-echo sequence with a resolution of 2 x 0.31 x 0.31 mm3 and 3D image reconstruction. In the men, the absolute volumes of the femur and tibia, but not those of the patella, were significantly higher than in the women. The differences between the sexes were considerably lower after normalisation to the body weight and the tibial head diameter and were no more statistically significant. The interindividual variability was reduced after normalisation to these two parameters, the body weight being more effective. We did not observe sex-specific differences in the percentage of the total cartilage volume taken up by the various joint surfaces. Our results suggest that, in young individuals without cartilage lesions, there exist sex-specific differences of the cartilage volume in the knee joint. However, these can be explained in terms of general differences in body constitution (body weight and bone size), without further significant influences of the sex. The knowledge of the normal, sex-specific cartilage volume is relevant when attempting to estimate the amount of tissue loss at the time at which symptoms occur in a patient with degenerative joint disease. PMID- 9862028 TI - Morphological and histochemical ageing changes in patellar articular cartilage of the rat. AB - The aim of this study was to investigate the variation in cartilage characteristics with age. Fresh-frozen cryostat sections of the patellar articular cartilage of the rat were used to demonstrate the enzyme activity of succinate dehydrogenase, lactate dehydrogenase, alkaline phosphatase, and acid phosphatase in the different layers and at different ages. Light microscopic techniques were used to analyse quantitative features such as thickness, cell density and the histological characteristics of the articular cartilage. The results indicate that cell density is significantly affected by age. Furthermore, it depends on the distance from the surface. The most marked decline in cell density occurred between months 3 and 6. The thickness of the articular cartilage also varies with age. The reduction in cartilage thickness was most striking between months 3 and 6. Differentiation into the histological layers is obvious after 3 months. Glycolytic enzymes were strongly reactive in all regions and at all ages, whereas aerobic activity declines with age. The metabolic and morphological changes in ageing cartilage contribute to trophic disorders and deterioration of the functional cartilaginous situation in adult cartilage. PMID- 9862029 TI - Ultrastructural aspects of cartilage formation, mineralization, and degeneration during primary antler growth in fallow deer (Dama dama). AB - Due to their rapid growth, regular replacement and easy accessibility, deer antlers are considered a useful model for the study of cartilage and bone differentiation and mineralization in mammals. The present study describes, for the first time, the cellular and extracellular matrix changes associated with cartilage formation, mineralization and degeneration in primary antlers on the ultrastructural level. Growing primary antlers of 3 to 4 cm length were obtained from six fallow bucks, aged about 10 months. It was shown that the chondroblasts were derived from progenitor cells of the antler perichondrium and differentiated into mature chondrocytes that subsequently underwent hypertrophic changes. Concomitant with cell hypertrophy, formation of a lacunar and a perilacunar extracellular matrix was observed, the latter containing numerous collagenous fibers. Mineralization of the extracellular matrix occurred via matrix vesicles and the formation of apatite crystals at distinct sites of the collagenous fibers. The hypertrophic chondrocytes of the mineralized cartilage then degenerated, a process that was also occasionally observed in more distally located cells surrounded by still unmineralized matrix. No morphological indications of a transdifferentiation of hypertrophic chondrocytes into bone forming cells, i.e., co-occurrence of a degenerating chondrocyte and a viable osteogenic cell in intact lacunae, were found. The cellular and extracellular matrix changes seen in primary antlers resemble those described for secondary antlers. Our results further indicate that the hypertrophic chondrocytes of primary antlers eventually undergo apoptosis, thereby providing further evidence that metaplastic conversion of cartilage into bone does not play a role in antler growth. PMID- 9862030 TI - Effects of prostaglandin E2 on growth, morphology, morphometry and keratin pattern of bovine corneal epithelial cells cultured in vitro. AB - In this work the relationship between the proliferation of bovine corneal epithelial cells and PGE2 has been studied. Our data indicate that PGE2 plays an important role in the growth of corneal epithelial cells. Actually, epithelial cells cultured on a keratocyte feeder-layer and exposed to indomethacin, a cyclooxygenase inhibitor, have shown a decrease in growth rate at drug concentrations which otherwise did not induce a reduction in the viability of the keratocytes as well as in epithelial cells in separate cultures. This effect has been reversed by an exogenous PGE2 addition to the culture media. Moreover, significant increases have been found in the growth of epithelial cells cultured in the presence of keratocytes, with basal medium and with conditioning medium after adding exogenous PGE2 at concentrations equal to or lower than 10(-6) M. Significant decreases in the dimensions of the corneal epithelial cells have been found only when PGE2 has been added to basal and to conditioning medium, suggesting that the autacoid maintains cell dimension and morphology. The appearance of keratins with high molecular weight (54 and 57 kDa) coupled with the tendency to stratification of the cells cultivated with media supplemented with PGE2, indicates that the autacoid could favour cell differentiation. The action of PGE2 on the corneal epithelial cells does not seem to be influenced by the presence of the fibroblasts and their products, since PGE2 has induced increases in cell growth and morphological variations, independent of cultural conditions and therefore also only in the presence of basal medium. PMID- 9862031 TI - The structure and function of periodontal ligament cells in acellular cementum in rat molars. AB - To elucidate the structure and function of periodontal ligament cells at the periodontal ligament-cementum interface in advanced acellular cementogenesis, the cervical regions of molars in rats aged 6 weeks were observed by light and electron microscopy. The light and transmission electron microscopy showed the periodontal ligament cells to be elongated between dense, well-developed principal fibers. The transmission and scanning electron microscopy showed that these cells extended wing-like projections from the lateral surface, forming cylindrical compartments surrounding the principal fibers. In addition, finger like projections extended toward the cementum from the cementum-facing ends. The main results suggest the following: at the periodontal ligament-cementum interface, the periodontal ligament cells maintain the architecture of the principal fibers by means of extracellular compartments. The arrangement of finger-like projections results in the formation of acellular cementum containing only Sharpey's fibers as a fibrous component. PMID- 9862033 TI - Telencephalic connections of the visual system of the chicken: tracing the interrelation of the efferents of the visual Wulst and the hyperstriatum ventrale. AB - In the present study the telencephalic connections between the centres of the tectofugal and thalamofugal pathways were investigated with Phaseolus vulgaris leucoagglutinin and biotinylated dextran-amine anterograde tracers in chicken using light and electron microscopy. No direct connection was found between the visual Wulst and the ectostriatum or the telencephalic centres of the tectofugal and thalamofugal pathways. Besides other projections, the visual Wulst emitted fibres also to the middle and lateral parts of the hyperstriatum ventrale. Further experiments revealed that the middle part of the hyperstriatum ventrale projected to the ectostriatum centrale and periphericum and established an indirect connection between the visual Wulst and the ectostriatum. The lateral part of the hyperstriatum ventrale sent a few efferent fibres toward the diencephalon and brainstem, but projected massively to the ectostriatum periphericum, neostriatum intermedium pars laterale, the ventral part of the neostriatum caudale and the archistriatum dorsale. Considering that these areas are structures of the tectofugal circuitry as well, the presence of these connections may result in a more elaborate visual processing. The neostriatum may be an associative visual center and possibly a modulatory area toward the archistriatum intermedium dorsale, by which visual information may mediate, modulate and control the movements. PMID- 9862032 TI - Granin proteins (chromogranin A and secretogranin II C23-3 and C26-3) in the intestine of amphibians. AB - The occurrence, distribution and possible cellular colocalizations of chromogranin A (CgA) and of two synthetic secretogranin II peptides (SgIIC23-3 and SgIIC26-3) with serotonin, somatostatin, neurotensin, pancreatic polypeptide and bombesin have been investigated immunohistochemically in the amphibian gut. CgA or SgIIC26-3-immunostained enterocytes were found throughout along the frog intestine, while no immunoreaction for any of the tested antisera against granins was seen in the same organ of newts. Variable amounts of serotonin-immunoreactive cells co-storing CgA or SgIIC26-3, but never both granins, were encountered in all intestinal segments of the frogs investigated. In addition, CgA was co localized with somatostatin in a few endocrine cells of the frog (genus Rana) duodenum and small intestine. In the duodenum of another frog (genus Xenopus) several enterocytes co-stored SgIIC26-3 and neurotensin. Pancreatic polypeptide- and bombesin-immunoreactive cells, the latter detected only in the duodenum of Xenopus, did not contain and granin. The results suggest that, in spite of their relatively restricted occurrence in the intestine of frogs and even of their absence in that of newts, the granins are well conserved during phylogeny. On the other hand, the heterogeneous distributions of these anionic glycoproteins, related to the entero-endocrine cell types, make their previously assigned usefulness as markers of all neuro-endocrine cells unlikely. PMID- 9862035 TI - Angioarchitectural structure and functional distributive pattern of the filiform papillae on the meso-dorsal surface of the rabbit tongue. AB - The angioarchitectural classification and distributive patterns were investigated in the filiform papillae (FiP) on the meso-dorsal surface of the rabbit tongue by using microvascular cast specimens (MVCS) and the scanning electron microscope (SEM). The author examined the microvascular network structures, which consisted of ascending and descending branches entering the FiP. They inclined in a posterior (pharynx) direction, and densely and geometrically covered the meso dorsal surface, directing the spoon-like concave face in an anterior direction. FiPs could be classified into three types: a small filiform papilla (SfP) covering on the meso-dorsal surface except for the marginal part of the intermolar eminence (MIME) and the intermolar eminence itself (IME): a spoon-like concave structure facing in an anterior (apex) direction with an arrowhead-like top: a middle filiform papilla (MfP) on the MIME, made up of a long triangle-like concave structure with a sharp arrowhead-like top and inclined at right angles to the IME. A large filiform papilla (LfP) on the whole swelling dorsal area of the IME was formed by a long triangle-like concave structure with a sharper arrowhead like top. LfPs are longer and larger than MfPs and inclined towards the pharynx. PMID- 9862036 TI - Development of the cribriform plate and of the lamina mediana. AB - The development of the cribriform plate and lamina mediana was studied in macerated isolated ethmoid bones in specimens from late fetal life to the stage of its final shape (60 specimens). From fetal life to the first year of age, the ethmoid bone consisted of two separate symmetrical halves which had joined together by the end of the first year. Each half of the future ethmoid bone incorporated the superior, middle and occasionally also the supreme nasal concha. The ossification of the cribriform plate started in the new-born where it initially displayed a vertical position but became horizontal in the course of the first year. At the end of the first year both halves of the ethmoid bone had been united by the formation of the crista galli, lamina mediana and complete ossification of the cribriform plate. The lamina mediana reached its final shape by ten years of age. Each half of the ethmoid bone displayed furrows for the fila olfactoria in the region of the superior and occasionally also of the anterior part of the middle nasal concha. The furrows run in a postero-anterior direction. In the course of our investigations we found three cases where all three nasal conchae formed a unique block thus proving the common origin of these structures from the cartilaginous nasal capsule. PMID- 9862034 TI - The lingual epithelium of Salamandra salamandra: metamorphotic changes during its ontogenesis. AB - The development of the epithelia of the secondary tongue of Salamandra salamandra is described on the basis of light microscopic and scanning electron microscopic studies of defined developmental stages. A glandular area with radial ridges and furrows is formed anterior to the primary tongue during the larval phase. Epithelial cones--each a compact anlage of a gland lying in the furrows--displace the lamina propria. The glandular area grows upward and latero-caudad during metamorphosis and forms the secondary tongue by fusing with the primary tongue. Lumina within the gland anlage appear at the beginning of metamorphosis. They open as glandular tubules towards the oral cavity at the climax of metamorphosis. The epithelial lining becomes single layered and differentiates into gland cells. The glands are increasingly surrounded by fibers of the musculus genioglossus. At the orifice of the gland, the gland cells mingle with the multilayered epithelium of the surface of the tongue. This contains two types of goblet cells in addition to the villus-shaped covering cells which leave gaps for the taste buds. The goblet cells are formed before (type I) and during (type II) metamorphosis and replace the typical larval goblet cells. The new mushroom-shaped part of the secondary tongue is characterized by aborally running septae of connective tissue, visible after digestion with pankreatin. The tip of the primary tongue which originally covers the glandular part becomes completely integrated. It is characterized by crypts which become shallower caudally. PMID- 9862037 TI - Venous drainage of the stomach in the golden hamster (Mesocricetus auratus) and the guinea pig (Cavia aperea f. porcellus). AB - Venous drainage patterns of the stomach were studied in 30 golden hamsters (Mesocricetus auratus) and in 30 guinea pigs (Cavia aperea f. porcellus). In golden hamsters the most frequent group had a v. gastroepiploica dextra (85.6% of cases). The v. gastroepiploica sinistra was found in 63.3% of cases. In the second group, the venous drainage pattern was without a v. gastroepiploica dextra (13.3% of cases). The only constant venous channel from the stomach was the v. gastrica sinistra. Other veins draining the stomach were vv. gastricae (tributaries of the v. lienalis) and interorgan venous anastomoses with neighbouring organs. A venous arch along the curvatura ventriculi major was not observed in any of the cases. In the guinea pig, the first group with a v. gastroepiploica dextra accounted for 90.0% of cases while the second group without this vein represented 10.0% of cases. The only constant venous channel was, once again, the v. gastrica sinistra. Other veins of the stomach were similar to those in the golden hamster. A venous arch along the curvatura ventriculi major was found in 76.6% of cases. Some differences in the venous pattern between the two animal species can be explained by zoological differences in these mammals. The results obtained also stimulate interest in the study of possible variations in venous patterns of the stomach in man. PMID- 9862039 TI - An unusual course of the right renal artery associated with an anomalous inferior vena cava. AB - In a 55-year-old male cadaver, the inferior vena cava bifurcated at a level midway between the hilus and inferior pole of the right kidney. The narrower branch, on the right side, drained the right renal and hepatic veins, ascended as the normal inferior vena cava, passed through the caval opening of the diaphragm and drained into the right atrium. The wider branch on the left side drained the left renal vein and penetrated the right crus of the diaphragm as the azygos vein. This vein ascended in the posterior mediastinum and drained into the superior vena cava. The hemiazygos and accessory hemiazygos veins were absent. In addition to these anomalies, the right renal artery entered the hilus after coursing through the above-mentioned two branches at the bifurcation. No congenital anomalies of the heart or abdominal viscera and main arteries were seen. PMID- 9862038 TI - Venous drainage of the stomach in the domestic rabbit (Oryctolagus cuniculus f. domestica, breed large Chinchilla) and the domestic cat (Felis catus L. f. domestica). AB - Two basic patterns of venous drainage of the stomach were found in rabbits. In the first, which is highly prevalent (27 cases-90.0%), the v. gastroepiploica dextra is present while no such vein is present in the second one. Other venous channels are represented by the v. gastrica sinistra and by tributaries of the v. lienalis. In cats there are also two basic patterns of the stomach venous drainage. The first highly prevalent type without the v. gastroepiploica dextra was found in 23 cases (76.0%); in the second type this vein was present. In rabbits the v. gastroepiploica sinistra was observed in 25 cases (82.5%), in cats in only two cases (6.6%). The v. gastroepiploica dextra in the rabbit is usually a tributary of the v. portae, while in cats it is a tributary of the v. lienalis. In ten rabbits (33.3%) there was a venous arch around the curvatura ventriculi major, while in cats it was absent. The great difference in the stomach venous pattern in rabbits and cats can be explained by the different zoological position of these two kinds of animals. PMID- 9862040 TI - Pulmonary development in the fetal one-humped camel: morphometry. AB - The structural development of the fetal camel lung is described and quantified morphometrically in this report. During fetal development the lung weight and volume increased polynomially with body length. The correlation coefficient between lung weight and body length is +0.991, and between lung volume and body length is +0.995. Volume density of the future bronchial tree showed a progressive increment with gestational age and is relatively higher in male than female fetuses, and the differences are statistically significant at 340-420 mm CRL. The proportion of lung tissue occupied by saccular airspaces increased enormously during the canalicular and alveolar stage, and the differences between both sexes are statistically significant at the former stage. The number of alveoli per unit area correlates closely with body length (r = +0.928) and the correlation coefficient between alveolar diameter and body length is +0.991. On the other hand, the correlation coefficient between the number of alveolar ducts per unit area and body length is -0.993, and is about +0.977 between the diameter of alveolar ducts and body length. PMID- 9862041 TI - New Drug Development Office (NDDO) splits from EORTC. PMID- 9862043 TI - Primary chemotherapy: a better overall therapeutic option for patients with breast cancer. PMID- 9862044 TI - Gene therapy of malignant glioma: recent advances in experimental and clinical studies. AB - Recent advances in molecular tumor biology and gene technology have provided the possibility to treat patients with malignant brain tumors by altering gene expression in tumor cells. Tumor development and progression involves alterations in a wide spectrum of genes, therefore a variety of gene therapy approaches for malignant gliomas have been proposed. In this review article, we discuss some principles of current gene therapeutic strategies that are under investigation in laboratories and in clinics. In addition, some general issues that remain to be resolved for clinical application of gene therapy in patients with malignant gliomas will be addressed. PMID- 9862045 TI - Critical reviews of economic analyses in order to make health care decisions for cancer. PMID- 9862048 TI - Thoracic metastasectomy for germ cell tumours: long term survival and prognostic factors. AB - BACKGROUND: This study evaluated the results of thoracic metastasectomy for germ cell tumours to assess long term survival and identify prognostic factors. PATIENTS AND METHODS: A series of 141 consecutive patients who underwent resection of thoracic metastases at Royal Brompton Hospital were retrospectively reviewed. Kaplan-Meier estimates of survival were calculated for clinical variables related to primary tumour and thoracic metastases, using the Cox model for multivariate analysis. RESULTS: Complete resection was achieved in 123 cases (87%); pathology showed viable malignant elements in 46 (32%), necrosis or fibrosis in 32, differentiated teratoma in 63. The overall survival was 77% at five years and 65% at 15 years, being significantly shorter in patients with malignant teratomatous elements (51% at five years, P = 0.0001) or incomplete resection (64% at five years, P = 0.019). At multivariate analysis these factors retained their prognostic value, with a relative risk of death of 5.7 for malignant teratomatous elements and 4.0 for incomplete resection. In addition, the Cox model revealed a 3.2 times higher risk of relapse in patients with malignant teratomatuos elements at the time of thoracic metastasectomy. CONCLUSIONS: These data confirm the value of thoracic metastasectomy to asses pathological response and achieve permanent cure of chemoresistant disease. PMID- 9862047 TI - A reduction in the requirements for mastectomy in a randomized trial of neoadjuvant chemoendocrine therapy in primary breast cancer. AB - BACKGROUND: A prospective randomised trial was undertaken to evaluate the role of neoadjuvant chemoendocrine therapy prior to surgery in primary operable breast cancer. PATIENTS AND METHODS: Three hundred nine women (median age 56 years, range 27-70) with primary operable breast cancer confirmed on fine needle aspiration (FNA) cytology were recruited to this study. They were treated with a combination of mitozantrone and methotrexate (+/- mitomycin-C) combined with tamoxifen (2MT). Patients received eight cycles of 2MT (four prior to surgery in the neoadjuvant group) and tamoxifen for five years with appropriate surgery and radiotherapy. The two groups were comparable for age, menopausal status, stage and surgical requirements. RESULTS: The clinical response rates to neoadjuvant therapy were as follows: 22% complete response (CR), 29% minimal residual disease (MRD), 33% partial response (PR), 15% no change (NC) and only two patients had clinical evidence of progressive disease. Surgical requirements were reduced from 31 patients (22%) of the adjuvant group having mastectomy to 14 (10%) in the neoadjuvant group (P < 0.003). At a median follow-up of 48 months (range 10-70 months) there is no statistically significant difference between the two groups in terms of local relapse, metastatic relapse or overall survival. Symptomatic and haematologic acute toxicity was low and similar for adjuvant and neoadjuvant therapy. CONCLUSION: This randomised trial has shown a significant reduction in the surgical requirements for mastectomy, after treatment with neoadjuvant chemoendocrine therapy, with no deterioration in local or distal relapse. PMID- 9862046 TI - Staging of breast cancer: what standards should be used in research and clinical practice? AB - BACKGROUND: Bone scan (BS), chest X-rays (CXR), liver ultrasonography (LUS) and laboratory parameters (LP) are frequently used as routine staging procedures for breast cancer patients. These procedures are not always appropriate in either clinical or research settings, regardless of the stage. The aim of this study was to identify groups of patients with differing risks for metastases in order to select more precise standard staging procedures. PATIENTS AND METHODS: The staging data relating to 406 breast cancer patients consecutively referred to our institution between November 1989 and October 1996 were analysed including pathological TNN grading and biological parameters. All of the cases with a positive or suspicious pre-operatory staging and who proved to have metastatic disease before surgery or during the first six months of follow-up were considered true-positive; all of the other cases with a positive or suspicious initial staging but with no evidence of distant metastasis before surgery and with a disease-free survival longer then six months were considered false positive. In the same way all cases with negative initial staging who relapsed during the first six months of follow-up were considered false-negative and those with negative initial staging and with a disease-free survival longer then six months were considered true-negative. Statistical analysis was performed using Fisher's exact test. RESULTS: BS, CXR and LUS, 388, 399 and 398 examinations respectively, were considered available, and 17 (4.38%), six (1.5%) and four (1%), respectively, proved to be true-positive. A statistically significant difference was observed when our cases were grouped according to T status (T4 vs. T1-T2-T3, P < 0.01) and nodal status (N0-N1 cases with less than three involved nodes and N1 with more than three positive lymph nodes N2 patients, P < 0.01). CONCLUSIONS: The present study suggests that breast cancer patients can be divided into three subgroups with different detection rates for distant metastases at staging (0.59%, 2.94% and 15.53%), and that the standard practice should be changed. In the first (T1N0 and T1N1 patients with < or = 3 positive lymph nodes--41.13% of the patients) and the second group (T2N0, T2N1 with < or = 3 positive lymph nodes, T3N0 and T3N1 patients with < or = 3 positive lymph nodes -33.49% of the patients) there is no need for a complete set of staging procedures, whereas full procedural staging is needed in the third group of patients (T4, N1 with > 3 lymph nodes and N2, 25.37% of the patients). PMID- 9862049 TI - Potential use of FDG-PET scan after induction chemotherapy in surgically staged IIIa-N2 non-small-cell lung cancer: a prospective pilot study. The Leuven Lung Cancer Group. AB - BACKGROUND: Clearance of viable tumour cells in mediastinal lymph nodes (MLN) by induction chemotherapy (IC)--so-called MLN downstaging--is an important aspect of combined-modality treatment of N2-NSCLC. Reassessment of MLN after IC by CT is far from accurate, while re-mediastinoscopy is often technically difficult. Based on our previous results with FDG-PET in the initial staging of N2 disease, we investigated whether PET after IC could be helpful in predicting MLN downstaging and therapeutic outcome. PATIENTS AND METHODS: Patients underwent a first PET before IC. After three cycles of platinum-based IC, a second PET was performed before locoregional therapy, either surgery or radiotherapy. PET results were correlated with pathology of the MLN when available, and with survival. RESULTS: Fifteen surgically staged N2-NSCLC patients were prospectively included. Locoregional therapy after IC consisted of surgery in nine and radiotherapy in six. Correlation with pathology of the nine resection specimens revealed that the accuracy of PET in predicting MLN downstaging was 100% (six true negatives; three true positives), whereas for CT it was only 67% (two false pos; one false neg). Reassessment with PET after IC was correlated with the outcome after the entire combined modality treatment. Survival was significantly better in patients with mediastinal clearance (P = 0.01) or with a greater than 50% decrease in the Standardised Uptake Value (SUV) of the primary tumour (P = 0.03) after IC. CONCLUSIONS: Mediastinal PET after IC accurately assesses pathologic MLN downstaging in N2-NSCLC. The data suggest a possible correlation of early survival with mediastinal clearance and an important decrease of SUV in the primary tumour. Confirmation of these preliminary findings would establish PET as a useful non-invasive tool to select patients for intensive locoregional treatment after IC. PMID- 9862050 TI - A phase II study of irinotecan alternated with five days bolus of 5-fluorouracil and leucovorin in first-line chemotherapy of metastatic colorectal cancer. AB - PURPOSE: This multicenter phase II study was designed to assess the efficacy of the alternating schedule of irinotecan (CPT-11) with bolus 5-fluorouracil (5-FU) and leucovorin (LV) in first-line chemotherapy for metastatic colorectal cancer (CRC). PATIENTS AND METHODS: Patients with histologically proven metastatic colorectal cancer, and at least one bidimensionally measurable lesion, aged 18 70, with performance status < or = 2, normal baseline biological values and no prior chemotherapy (or only adjuvant chemotherapy completed > or = 6 months before study entry) were selected. Treatment was irinotecan 350 mg/m2, i.v., day 1, alternating with leucovorin 20 mg/m2 i.v. and 5-FU 425 mg/m2, i.v. daily for five consecutive days, day 22-26 (Mayo Clinic regimen). One alternating cycle was to be performed every six weeks. Patients were evaluated for efficacy every alternating cycle. Treatment was administered until five alternating cycles, disease progression, unacceptable toxicity or patient refusal. RESULTS: Thirty three patients (28 chemotherapy-naive and five with prior adjuvant treatment completed > 1 year prior to accrual) were enrolled. The objective response rate (RR) was 30% (95% CI: 16-49; 10 patients/33; nine partial response and one complete response). All responses were reviewed by an independent external review committee. An additional 49% of patients had stable disease. The median survival was 16 months, the one year survival amounted to 58% and the median progression free survival was 7.2 months. Relative dose intensity was nearly 90% for both drugs. Grade 3-4 diarrhea and neutropenia were the most frequent severe toxic events, seen in 24% and 64% of patients, respectively. CONCLUSIONS: The alternating schedule of CPT-11 350 mg/m2 with five days bolus of 5-FU and low dose LV is an active and feasible regimen as front-line therapy for metastatic CRC. It is well tolerated, without evidence of overlapping toxicity. The response rate appears promising with regard to that expected with either single agent. This regimen warrants further assessment in randomized trials. PMID- 9862051 TI - A randomised, concentration-controlled, comparison of standard (5-day) vs. prolonged (15-day) infusions of etoposide phosphate in small-cell lung cancer. AB - PURPOSE: This randomised trial was designed to investigate the activity and toxicity of continuous infusion etoposide phosphate (EP), targeting a plasma etoposide concentration of either 3 micrograms/ml for five days (5d) or 1 microgram/ml for 15 days (15d), in previously untreated SCLC patients with extensive disease. PATIENTS AND METHODS: EP was used as a single agent. Plasma etoposide concentration was monitored on days 2 and 4 in patients receiving 5d EP and on days 2, 5, 8 and 11 in patients receiving 15d EP, with infusion modification to ensure target concentrations were achieved. Treatment was repeated every 21 days for up to six cycles, with a 25% reduction in target concentration in patients with toxicity. RESULTS: The study has closed early after entry of 29 patients (14 with 5d EP, 15 with 15d EP). Objective responses were seen in seven of 12 (58%, confidence interval (CI): 27%-85%) evaluable patients after 5d EP, and two of 14 (14%, CI: 4%-42%) evaluable patients after 15d EP (P = 0.038). Grade 3 or 4 neutropenia or leucopenia during the first cycle of treatment was observed in six of 12 patients after 5d EP and 0/14 patients after 15d EP (P = 0.004), with median nadir WBC count of 2.6 x 10(9)/1 after 5d and 5.0 x 10(9)/1 after 15d EP (P = 0.017). Only one of 49 cycles of 15d EP was associated with grade 3 or worse haematological toxicity, compared to 14 of 61 cycles of 5d EP. CONCLUSIONS: Although the number of patients entered into this trial was small, the low activity seen at 1 microgram/ml in the 15d arm suggests that this concentration is below the therapeutic window in this setting. Further concentration-controlled studies with prolonged EP infusions are required. PMID- 9862052 TI - Addition of etoposide to CHOP chemotherapy in untreated patients with high-grade non-Hodgkin's lymphoma. AB - BACKGROUND: Second- and third-generation chemotherapy protocols for the treatment of aggressive non-Hodgkin's lymphomas (NHL) have considerable, and age-related, toxic effects. In addition, they do not seem to prolong overall survival in comparison to standard CHOP chemotherapy. In this phase II study we investigated the feasibility and efficacy of the addition of etoposide to the conventional CHOP regimen. PATIENTS AND METHODS: Toxicity and clinical efficacy were determined in 132 patients with previously untreated high-grade NHL. There were 51 patients in clinical stage I and II and 81 patients in stage III and IV, with a median age of 54 years (range 17-85). Patients received standard-dose CHOP plus etoposide 100 mg/m2 i.v. on day 1 and 200 mg/m2 p.o. on days 2-3. RESULTS: The overall response rate was 84%, with 70% complete and 14% partial responses. The predicted three- and five-year survivals for the group as a whole were 60% and 53%, respectively, and the corresponding disease-free survivals for patients achieving complete remissions were 65% and 56%, respectively. Outcome was not different from that of CHOP-treated patients in a recently completed Nordic study performed during the same time period. Myelosuppression (WHO grade 3-4), observed in 87% of patients and infectious complications (WHO grade 3-4) in 33%, dominated the toxicity profile of this regimen. Fifty-seven of 92 complete responders (62%) received 6-8 CHOP-E cycles with no reductions in planned dose intensity. LDH level higher than normal, extranodal sites = 2, stage III-IV at diagnosis were all indicators of a poor survival. CONCLUSIONS: We conclude that CHOP-E treatment is effective in high-grade NHL. However, mainly due to severe myelosuppression frequent schedule modifications were required and the results are not obviously superior to those of conventional CHOP. PMID- 9862054 TI - Descriptive epidemiology of vulvar and vaginal cancers in Vaud, Switzerland, 1974 1994. AB - BACKGROUND: To analyse trends in incidence, survival and risk of second neoplasms following vaginal and vulvar cancers using data collected by the Swiss Cancer Registry of Vaud over the 21-year period 1974-1994. MATERIALS AND METHODS: Subjects were 257 vulvo-vaginal cancers. Of these, 69 were vaginal, 153 vulvar cancers, and 35 non-specified lower genital tract neoplasms; 94 in situ neoplasms were also registered (85 for the vulva). RESULTS: Invasive vaginal cancer incidence decreased from 0.8 in 1974-1984 to 0.4/100,000 women in 1985-1994, while invasive vulvar cancer incidence remained approximately stable around 1.2/100,000 (world standard); incidence of in situ vulvar cancer increased from 0.8 to 1.3/100,000, the rise being larger in younger women. Significant excesses for second primary neoplasms were observed for pro-pharyngeal and lung cancer, and for non-melanomatous skin neoplasms, as well as for invasive vulvar cancers following in situ cancers. CONCLUSIONS: This population-based dataset confirms that the incidence of in situ vulvar (but not invasive vulvar or vaginal cancer) has been increasing over the last 20 years. The excess second primary neoplasms supports the hypotheses that human papillomavirus and cigarette smoking are related to vulvo-vaginal neoplasms. PMID- 9862053 TI - Elderly patients with non-Hodgkin's lymphoma: population-based results in The Netherlands. AB - BACKGROUND: To compare characteristics, treatment and outcome of patients > or = 70 years with patients < 70 years in a population-based non-Hodgkin's lymphoma (NHL) registry. PATIENTS AND METHODS: All new patients with NHL (n = 1168) in a geographically defined region in the western part of The Netherlands were registered during a nearly 10-year period. Patient, tumour and treatment characteristics, response to therapy and survival were analysed for both age groups. An age-adjusted prognostic index was determined for elderly patients with aggressive lymphoma. RESULTS: The elderly comprised 41% of the registered patients. There were significantly more females, a preponderance of intermediate grade histology (diffuse large B-cell lymphoma) and a lower performance status. Incomplete staging in the elderly was mostly due to the omission of a bone marrow biopsy. With respect to WF grading the complete remission rate (except for patients with low-grade/stage I NHL, patients with extranodal NHL and for patients with intermediate grade/extensive NHL) and overall survival at five years (except for patients with low-grade/stage I NHL and for patients with intermediate-grade/extensive NHL) were significantly inferior in the elderly. With respect to the R.E.A.L. Classification the exceptions were in patients with high grade MALT lymphomas (elderly good) and patients with mantle-cell and peripheral T-cell lymphomas (younger group bad too). However, once complete remission was reached, the disease-free survival did not differ significantly between the two age groups, emphasising the importance of achieving complete remission. Although 65% of the classified elderly patients presented with intermediate-grade NHL, only 26% of the elderly patients treated with chemotherapy received anthracycline-based chemotherapy. In the elderly, lymphoma (treatment-related toxicity included) contributed to death in 70% and concomitant disease (other malignancy included) in 30%, versus 78% and 22%, respectively, for the younger group (P = 0.04). The age-adjusted prognostic index, made up of the factors serum LDH, stage and Karnofsky index, showed a clear distinction between the four risk categories low, low/intermediate, intermediate/high and high, with a median survival time of 43, 20, seven and four months, respectively. For the younger group the respective numbers were 144, 45, 19 and 11 months. CONCLUSIONS: In a population-based NHL registry the elderly, predominately female patients, formed a larger proportion of the patient group than the one usually reported in the literature. In this population-based cohort inferior remission and overall survival rates were seen in the elderly. However, obtaining complete remission was beneficial for the prognosis of this disease in the elderly. By the application of the R.E.A.L. Classification important subgroups emerge. PMID- 9862055 TI - Ventricular arrhythmia and torsade de pointe: dose limiting toxicities of the MDR modulator S9788 in a phase I trial. AB - BACKGROUND: S9788 is a triazineaminopiperidine derivative capable of reversing multidrug resistance (MDR) in vitro. In preclinical models S9788 was several fold more potent MDR inhibitor than verapamil or cyclosporine. At P-glycoprotein (Pgp) blocking concentrations, S9788 appeared to have only very little toxicity. PATIENTS AND METHODS: In a two step phase I trial we treated 39 patients with refractory cancer with S9788 and bolus doxorubicin. The steps differed mainly in the S9788 infusion duration; in the first part 23 patients received the MDR reversing drug S9788 over 30 minutes, in the second step of the study 16 patients were administered S9788 over 150 minutes. The doses of S9788 were escalated in cohorts of three patients up to a dose level (DL) of 96 mg/m2 on the 30 minutes infusion, and to 144 mg/m2 on the 150 minutes infusion. The pharmacokinetics of S9788 were determined. RESULTS: With the 30-minute infusion schedule symptomatic cardiac arrhythmia were found to be dose limiting. In all patients at the highest DL transient cardiac repolarization prolongation with a long QT-interval on ECG was demonstrated. With the 150-minute administration schedule, S9788 could be escalated up to 144 mg/m2 without subjective toxicity. However, transient QT prolongation was present in all patients. A third degree AV-block and a QT increase of about 40% occurred at the highest DL. Asymptomatic torsade de pointe (DL 96 mg/m2) was demonstrated on Holter recording in one patient. Theses repolarization disturbances with QT increase were considered dose limiting toxicity and the trial was closed. No arrhythmia related death was noted. Pharmacokinetics were similar with both infusion schedules with a mean alpha half life of 11.3 and 13.2 minutes, for the 30-minute and 150-minute infusion, and a terminal half life of 13.5 and 15 hours, respectively. QTc prolongation duration appeared to be dose-dependent. CONCLUSIONS: With the tested infusion schedules, cardiac toxicity, in particular AV-blocks and QT prolongation, leading to ventricular arrhythmia and torsade de pointe, are the dose limiting toxicities of S9788. Our experience together with the observation of asymptomatic torsade de pointe in two other phase 1 trials of S9788 infused over six hours precluded the further clinical development of S9788. PMID- 9862058 TI - Bimonthly high-dose leucovorin, 5-fluorouracil infusion and oxaliplatin (FOLFOX3) for metastatic colorectal cancer resistant to the same leucovorin and 5 fluorouracil regimen. AB - BACKGROUND: FOLFOX2, a bimonthly regimen of high-dose leucovorin (LV), 48-hour continuous infusion of 5-fluorouracil (5-FU) (LV-5-FU) and oxaliplatin (100 mg/m2) produced a high response rate (46%; 95% confidence interval (95% CI): 31% 60%) in 5-FU pre-treated patients with metastatic colorectal cancer. In this phase II study, pre-treated patients were given a lower dose of oxaliplatin to reduce the toxic effects of the regimen. PATIENTS AND METHODS: Thirty patients with advanced colorectal adenocarcinoma and progression while receiving bimonthly LV-5-FU (LV: 500 mg/m2, 5-FU: 1.5-2 g/m2/22 hours, days 1-2, every two weeks), were given the same LV-5-FU schedule with the addition of oxaliplatin (85 mg/m2) every two weeks (FOLFOX3). RESULTS: The main toxic effects were peripheral neuropathy (90%) with four severe sensitive neuropathies (WHO grade 2: 13%). The response rate was 20% (95% CI: 8%-39%). Median progression-free survival was 26 weeks, median survival was 57 weeks from the start of FOLFOX3 and median duration of the response was 37 weeks. CONCLUSIONS: Results obtained with FOLFOX3 confirmed the synergy between oxaliplatin and 5-FU in 5-FU-resistant metastatic colorectal cancer. However, the response rate seems to be lower than that obtained with FOLFOX2. Further studies to determine the best oxaliplatin dose intensity are in progress. PMID- 9862059 TI - Prognostic impact of an activation of coagulation in lung cancer. PMID- 9862057 TI - A phase I-II study of bi-weekly paclitaxel as first-line treatment in metastatic breast cancer. AB - BACKGROUND: Single-agent bi-weekly paclitaxel was studied as first-line metastatic treatment for breast cancer in a phase I-II trial. PATIENTS AND METHODS: Thirty-eight women with metastatic breast cancer were enrolled. Thirty seven are evaluable for toxicity, 35 for response. RESULTS: The MTD was defined at 160 mg/m2 q two weeks with dose limiting toxicity in two patients consisting of hematological toxicity (1) and neurotoxicity (2). Twenty patients were treated at 150 mg/m2, the recommended dose. Response rates were two CRs and nine PRs (overall 61%) at the RD of 150 mg/m2 and three CRs and 11 PRs for an overall RR of 67% for the two top doses. CONCLUSIONS: The good drug tolerance, response rates, and convenience over weekly treatment suggest this may be a worthwhile regimen. PMID- 9862060 TI - Myotonia associated with hyperadrenocorticism in two dogs. AB - Two dogs developed a disabling gait abnormality characterised by stiffness. The abnormality was consistent with a diagnosis of myotonia secondary to hyperadrenocorticism. The first dog had iatrogenic hyperadrenocorticism, and its signs improved substantially after corticosteroid administration was gradually withdrawn. The second had pituitary-dependent hyperadrenocorticism, but myotonic signs progressed despite effective mitotane therapy. Procainamide administration reduced the myotonic stiffness in the second case. PMID- 9862056 TI - Abdominal pain with anorexia in patients with breast carcinoma. AB - Breast cancer is the second commonest primary tumour responsible for gastrointestinal metastases after malignant melanoma. The real incidence of gastrointestinal metastases in breast cancer patients is probably underestimated owing to the non-specific presenting symptoms and death of patients caused by other more obvious metastases. The predominant histological subtype of gastrointestinal metastases of breast cancer is invasive lobular carcinoma and the median interval from diagnosis of primary breast cancer to gastrointestinal metastases is five years. We report two cases of disseminated breast cancer with gastrointestinal involvement with a rather long survival. PMID- 9862061 TI - Clinical and anatomical features of lymphosarcoma in 118 cats. AB - OBJECTIVE: To determine patients' characteristics and anatomical distribution of lesions in cats with lymphosarcoma. DESIGN: Prospective multi-institutional study of naturally occurring feline lymphosarcoma. METHODS: Veterinarians in Sydney were provided with free diagnostic laboratory services for suspect cases of feline lymphosarcoma. Lymphosarcoma was diagnosed based on physical findings, radiographic and/or ultrasonographic images and results of cytological or histopathological examination. When owners were not interested in pursuing an antemortem diagnosis, suspect cases were collected for necropsy. Patients' characteristics and physical findings were recorded. A modified scheme for anatomical classification of lesions was devised including a 'mixed' category for cases which involved two or more anatomical forms. RESULTS: One hundred and eighteen cases were accrued over an 18 month period. The median age was 120 months and range 5 to 212 months. Age distribution was bimodal, with a small peak for cats less than 24 months, and a normal distribution centred on 97 to 120 months. Eighty cats were domestic crossbreds, 22 were Siamese or Oriental cats (including crosses), 6 were Burmese, 5 were purebred longhairs and the remaining 5 were one of a number of purebred shorthaired breeds. In comparison to 1017 consecutive cases admitted to our hospital for conditions other than lymphosarcoma, Siamese/Oriental cats were over-represented amongst lymphosarcoma cases (P = 0.0006). Male cats were also over-represented, accounting for 72 of 118 cases (P = 0.05). Abdominal lymphosarcoma was the most common anatomical form (43 cats), followed by mixed (39), nodal (20), mediastinal (9) and atypical (involving non-lymphoid organs, 7) forms. When analysed for specific organ involvement, 29 (25%) had mediastinal involvement, 71 (60%) had abdominal involvement including 60 (51%) with involvement of the intestinal tract and/or mesenteric lymph nodes and 36 (31%) with bilateral renal involvement, and 47 (40%) had peripheral lymph node involvement. No case of primary lymphoid leukaemia was identified. A noticeable subgroup of cats younger than 24 months had involvement of the anterior mediastinum with or without concurrent enlargement of cervical or axillary lymph nodes; Siamese/Oriental cats were over represented in this subgroup. Among cases with nodal involvement, lymph nodes of the head and neck were frequently involved, mandibular nodes most commonly, followed by superficial cervical nodes. In seven cases a solitary node was affected. CONCLUSIONS: Compared with similar surveys overseas, our cats were older and male cats were over-represented. There was a notable subgroup of young cats with mediastinal involvement. Siamese/Oriental cats were over-represented in this subgroup as well as in the larger population of cats with lymphosarcoma. Compared with overseas surveys, renal involvement, mixed cases and atypical cases (including nasal lymphosarcoma) were more common. A new subcategory of nodal lymphosarcoma, with involvement restricted to node(s) of head and neck, was identified. PMID- 9862063 TI - Dealing with diseases in wildlife. PMID- 9862062 TI - Progressive tetraparesis and laryngeal paralysis in a young rottweiler with neuronal vacuolation and axonal degeneration: an Australian case. AB - A 5-month-old female Rottweiler dog was diagnosed to have a neurodegenerative disease that has been recently reported in Rottweilers from North America and Europe. The dog was presented with progressive signs of ataxia, tetraparesis and inspiratory stridor. The clinical investigation included analysis of CSF, radiography, myelography and electrophysiological testing. No evidence of vertebral malformation or inflammatory CNS disease was identified. Bilateral laryngeal paralysis was identified in the lightly anaesthetised dog. Electromyography showed abnormal spontaneous activity from the intrinsic musculature of the larynx. At necropsy there were no gross abnormalities of the nervous system but there was atrophy of the dorsal cricoarytenoid muscles of the larynx. There were widespread histological abnormalities throughout the nervous system including neuronal vacuolation, spongiform changes in the neuropil and axonal degeneration which was most prominent in the spinal cord. These clinical and pathological findings are consistent with the diagnosis of a new neurodegenerative disease reported from North America and Europe. This diagnosis is of particular significance in Australia where transmissible spongiform encephalopathies have not been identified. PMID- 9862066 TI - Attitudes and practices of Queensland dairy farmers to the control of the cattle tick, Boophilus microplus. AB - OBJECTIVE: To determine practices for the control of cattle ticks on dairy farms in Queensland, the attitudes of farmers to tick infestations and to identify opportunities for and barriers against the introduction of non-chemical methods of tick control. DESIGN: A survey of 199 dairy farmers from tick-infested parts of Queensland was undertaken by 20 dairy advisers and stock inspectors from October 1996 to June 1997. The sample was a proportional, random selection of dairy farms from four regions. A personal interview was conducted with each farmer and answers to 134 questions were obtained. RESULTS AND CONCLUSIONS: Most farmers were not concerned by cattle ticks on their own farms, although they believed that ticks are important to the dairy industry. They were most concerned about the development of chemical resistance by cattle ticks. Inadequate facilities and lack of motivation appeared to be the factors most limiting to improving the methods of control. Most farmers claimed to have only small numbers of ticks at worst. Although a control program recommended by the Queensland Dairyfarmers' Organisation was well regarded by farmers, few had adopted it. Many farmers saw no need to implement a strategic control program. PMID- 9862064 TI - Pesticide use and residues on Queensland wool. AB - OBJECTIVE: To determine practices for control of louse infestation and blowfly strike in Queensland sheep flocks that are associated with organophosphorous and synthetic pyrethroid residues on wool. DESIGN: Information on residues was obtained from a survey of Queensland wool clips. Information on pesticide use was obtained from a trace-back postal survey. The association between pesticide use and residues was assessed using generalised linear models, controlling for potential confounding by flock location. PROCEDURE: Between 1995 and 1997 Queensland wool clips were randomly sampled. Samples were tested for the presence and amount (mg per kg of greasy wool) of organophosphorous and synthetic pyrethroid pesticides. A questionnaire seeking information on flock characteristics and pesticide use was sent to the manager of each flock from which a wool sample was tested. RESULTS: The median amount of OP and SP residue was 0.8 and 0.25 mg/kg, respectively, and 91 and 95% of wool samples contained < 8 mg/kg of OP and SP residues, respectively. The frequency of OP pesticide use for louse control was significantly (P = 0.005) associated with mean OP residue amount, and the timing of SP use for louse control, in relation to shearing, was significantly (P < 0.001) associated with mean SP residue amount. CONCLUSION: Most Queensland wool clips have acceptable amounts of residues after the use of OP and SP pesticides, but wool growers can further reduce residues by effectively controlling louse infestation with pesticide applications early after shearing and the use of non-chemical methods of ectoparasite control. PMID- 9862065 TI - Skin disease affecting the conservation of the western swamp tortoise (Pseudemydura umbrina) AB - OBJECTIVE: To review the present position of the western swamp tortoise (Pseudemydura umbrina) as an endangered species and significant health issues affecting efforts to save it from extinction. PROCEDURE: A retrospective analysis of the husbandry, hospital and pathology records of the western swamp tortoise captive breeding program at Perth Zoo. RESULTS: In 1987 a captive breeding project was developed to prevent the extinction of the western swamp tortoise but an outbreak of a necrotising dermatitis in 1989 threatened the survival of the captive bred hatchlings. Less severe outbreaks occurred in 1990 and 1993, with isolated cases in between. Of 283 tortoises that were born in captivity or came into captivity from the wild, 37 (13.1%) were affected, comprising 37% of all males, 26% of all females and 13% of animals of unknown gender. Of the affected animals, 70% were less than 2 years of age and 29% were older. Males were 1.6 times more likely to be infected than females but this difference was not statistically significant (P = 0.27). Culture of the lesions consistently yielded unidentified Pseudomonas sp. CONCLUSION: Improved husbandry, such as strict maintenance of water quality and temperature conditions similar to that of the animal's natural habitat, and monitoring the health of individual tortoises have successfully controlled skin disease in the captive breeding of the western swamp tortoise. PMID- 9862067 TI - Development of a toxin-binding agent as a treatment for tunicaminyluracil toxicity: protection against tunicamycin poisoning of sheep. AB - OBJECTIVE: To assess the ability of certain derivatives of beta-cyclodextrin to treat sheep affected by tunicaminyluracil toxicity, using tunicamycin poisoning as a model system. DESIGN: Controlled treatment trial. ANIMALS: One hundred and sixty Merino wethers were used in the studies. PROCEDURE: Groups of sheep were experimentally poisoned with tunicamycin. Derivatives of beta-cyclodextrin, with or without magnesium sulphate and magnesium gluconate, were administered to treatment groups daily for 2 to 3 days. Treatment groups were compared with untreated groups in terms of survival. RESULTS: A significant increase in survival was observed following treatment of tunicamycin-affected sheep with hydroxypropyl-beta-cyclodextrin (HP beta-CD) and magnesium sulphate in solution (P < 0.05). In subsequent trials, formulation of the cyclodextrin in the form of a magnesium gluconate gel suspension demonstrated significant protection (P < 0.01) and was equally as effective as the cyclodextrin in solution, but required half the frequency of administration, even when the treatment was not commenced until 24 h after the final toxin dose. Beta-cyclodextrin-epichlorohydrin copolymer also improved the survival rate. After toxin administration, the sheep lost significantly less weight if treatment with HP beta-CD was commenced early (P < 0.001). CONCLUSION: Protection studies using these two beta-cyclodextrin derivatives suggest that they may be effective in increasing the survival of sheep poisoned by tunicamycin and warrant further testing in field outbreaks of annual ryegrass toxicity. PMID- 9862068 TI - Proliferative sparganosis in Australian dogs. PMID- 9862069 TI - An outbreak of mucormycosis in slender tree frogs (Litoria adelensis) and white lipped tree frogs (Litoria infrafrenata) PMID- 9862071 TI - Human behavioural pharmacology. PMID- 9862070 TI - Foot-and-mouth disease in Australia. PMID- 9862072 TI - Ethical and practical issues involved in behavioral pharmacology research that administers drugs of abuse to human volunteers. AB - Researchers carrying out non-therapeutic research that involves the administration of drugs of abuse to human volunteers can be faced with many ethical and practical questions. The history of this type of research is relatively brief, with little in the way of published information relevant to carrying out behavioral pharmacological research with human participants. The aim of this article is to raise issues that occur in most studies of this type and to provide solutions that we have found acceptable and which have been approved by a variety of institutions and regulatory agencies. Clearly, there are other approaches that would work equally as well, and we are not attempting to provide 'the' answer to many of the issues raised. We believe that raising these issues and providing our perspectives is important for stimulating others to discuss them and for all of us to strive, where possible, to reach a consensus concerning ethical practices and to become aware of gaps and pitfalls. The topics discussed range from the nuts and bolts of acquiring and keeping track of drugs, to selecting research participants and designing ethical studies that protect our volunteers while still collecting scientifically useful data. PMID- 9862074 TI - Ethical and theoretical paradoxes in human behavioral pharmacological research. Commentary on Fischman and Johanson's ethical and practical issues involved in behavioral pharmacology research that administers drugs of abuse to human volunteers. PMID- 9862075 TI - Homology in behavioural pharmacology: an approach to animal models of human cognition. AB - The distinction in biology between homology and analogy is examined for possible application to studies in behavioural pharmacology. It is argued that the concept of homology is central to understanding the 'construct validity' of animal models of human cognition. It is suggested that we capitalize on known correspondences across species in brain structure and development which may mediate homologous behavioural functions. Manipulation of specific receptors in defined areas may be achieved by local and systemic administration of drugs with relatively specific actions; this will complement an alternative criterion of construct validity, based on clinical treatments. This argument will be illustrated by a critique of successful extrapolations across species that are sometimes initiated by work in animals and sometimes by work in humans, especially in the clinical setting. The examples used will include analyses of spatial working memory, spatial attention and attentional set-shifting, studied in rats, monkeys and humans. PMID- 9862076 TI - Pharmacological approaches to animal models of human working memory and shifting attentional set. Commentary on Robbins' homology in behavioural pharmacology: an approach to animal models of human cognition. PMID- 9862077 TI - Mapping neuroanatomical correlates of the discriminative effects of drugs. Commentary on Robbins' homology in behavioural pharmacology: an approach to animal models of human cognition. PMID- 9862078 TI - What role models? Commentary on Robbins' homology in behavioural pharmacology: an approach to animal models of human cognition. PMID- 9862079 TI - Opioid discrimination in humans: discriminative and subjective effects of progressively lower training dose. AB - The purpose of this study was to examine the extent of covariation of subjective and discriminative drug effects as the dose of the discriminated training drug was progressively lowered. Six adult male volunteers with histories of opioid abuse, who were not currently physically dependent, were trained to discriminate the mu-receptor agonist hydromorphone (20 mg, oral) from placebo in daily sessions. They received financial reinforcement for correct responses. The hydromorphone training dose was then progressively reduced (20, 14, 10, 7, 5, and 3.5 mg) while the discrimination reinforcement contingencies remained in effect. Measures of subjective and physiological effects were concurrently collected during each discrimination session. As the training dose decreased, discriminative performance was generally well maintained, although the percent of drug-appropriate responses to hydromorphone did decline from 98% to 75%. The magnitude of the subjective and physiological effects of hydromorphone also decreased as the training dose decreased. At the lowest training dose, there were no physiological effects and few subjective effects of hydromorphone statistically different from placebo, although discrimination behavior remained statistically significant at all doses. These data indicate covariation of subjective effects and discrimination performance and suggest that discrimination behavior may be more sensitive for differentiating among drug conditions than traditional subjective effects measures. PMID- 9862080 TI - Zolpidem is differentiated from triazolam in humans using a three-response drug discrimination procedure. AB - The discriminative stimulus effects of the imidazopyridine hypnotic zolpidem and the classic benzodiazepine hypnotic triazolam were examined in seven healthy volunteers using a three-response drug discrimination procedure and a within subject design. During an initial sampling phase, the training drug conditions (placebo, 20 mg/70 kg zolpidem, and 0.5 mg/70 kg triazolam) were identified to subjects by letter codes before oral drug administration. During a subsequent training phase, subjects earned money for correct drug identifications made 3.75 h after drug administration. Five out of seven subjects acquired the three response discrimination. Analyses of standardized and unstructured self-report questionnaires revealed that zolpidem and triazolam produced different profiles of effects; zolpidem was associated with a number of negative somatic symptoms including nausea, blurred vision, visual images/hallucinations, and heavy limbs, whereas triazolam was associated with greater sedative effects. These results demonstrate a distinct profile of discriminative stimulus and subjective effects for zolpidem, relative to triazolam, which is consistent with its somewhat distinct pharmacological profile, and provide evidence for the sensitivity of the three-response drug discrimination procedure for detecting between-drug differences. PMID- 9862081 TI - Effects of the hallucinogen psilocybin on habituation and prepulse inhibition of the startle reflex in humans. AB - Schizophrenic patients exhibit deficits in indices of sensorimotor gating, such as habituation and prepulse inhibition (PPI) of the startle reflex. Hallucinogenic drug-induced states are putative models for the early and acute stages of schizophrenic and schizophrenia-spectrum disorders. Hallucinogenic drugs have been shown to disrupt PPI and/or retard habituation of the startle reflex in animal models of schizophrenia, consistent with the view of hallucinogen-induced states as 'model psychoses'. We evaluated the effects of the hallucinogen psilocybin on PPI and habituation of the startle reflex in a double blind, placebo-controlled human study with 12 healthy subjects. In contrast to animal studies, in our small human sample, psilocybin increased PPI, while having no clear effect on habituation (n = 6). These findings must be considered preliminary because several factors, including dose regimens and experimental parameters, may influence the results of studies on startle plasticity. Further investigations both with psychotic patients in different stages of the disease and with human and animal models of schizophrenia are needed in order to explore the effects of hallucinogens on sensorimotor gating and the relationship between information processing in hallucinogenic drug-induced states and the naturally occurring psychoses. PMID- 9862083 TI - Ketoconazole attenuates the cortisol response but not the subjective effects of smoked cocaine in humans. AB - Attenuation of hypothalamo-pituitary-adrenal (HPA) function in laboratory rodents has been found to reduce the reinforcing effects of cocaine. To examine whether attenuation of HPA function reduces the effects of cocaine in humans, one female and seven male 'crack' cocaine abusers were pretreated with three doses of ketoconazole (0, 600, 1200 mg), an inhibitor of adrenocorticoid biosynthesis, 1 h before receiving cocaine. Three doses of smoked cocaine (0, 12, 50 mg) were administered in counterbalanced order under each ketoconazole condition. Ketoconazole dose-dependently reduced cocaine-induced cortisol, but not adrenocorticotropin (ACTH) release, and attenuated the cocaine-induced increase in heart rate and blood pressure. Plasma ACTH levels were more predictive of blood pressure changes than either cocaine or cortisol levels. Suppression of cortisol secretion was not associated with a reduction in ratings of the subjective effects of cocaine. These results support a role for the HPA axis in the cardiovascular effects of cocaine, but do not support a role for the HPA axis in the subjective effects of cocaine. To the extent that self-administration can be predicted by subjective effects, these results further argue that the HPA axis does not play a critical role in cocaine self-administration by humans. PMID- 9862092 TI - FcR-mediated inhibition of cell activation and other forms of coinhibition. AB - The tripartite inactivation model proposed that coaggregation of the B cell antigen receptor (BCR) with the Fc receptor (FcR) by antigen and specific IgG antibody complexes explained the Fc-dependent inhibition of immune responses by antibody. This model has since been substantiated by many observations and its impact on studies of immune regulation has been threefold: (1) IgG antibody, via Fc gamma RIIB, mediates inhibition of cell activation in many cell types, demonstrating the general importance of this mechanism in immune regulation; (2) Fc gamma RIIB was the first receptor described that regulates immune responses by coinhibition, that is, regulation as a result of interaction between activating receptors (BCR, TCR, Fc epsilon RI, Fc gamma RIII, Fc gamma RIIA) and inhibitory receptors (Fc gamma RIIB, CTLA4, CD5, CD22, p58/70/140 KIR, gp49B1/gp91, Ly49A/C/E/F/G, NKG2-A/B, APCR, Fas (CD95), TGF beta-R, TNF-R, IFN gamma-R, and others). The list of coinhibitors is expanding, just as the list of costimulators has grown. Tolerance through multiple coinhibitors implies that Signal 1 alone is not tolerogenic; and (3) Studies of Fc gamma RIIB coinhibitory mechanisms have pointed the way to potential general inhibitory signaling pathways used by many receptors, involving the competing effects of various kinases and phosphatases, and other competitive events. Investigations of Fc gamma RIIB physiologic function and of other coinhibitory receptors, together with recent biochemical analyses, give an initial understanding of the biology of these inhibitory receptory receptors. Paradoxes within and between theoretical constructs, functional observations, and mechanistic studies point to critical questions for future study. PMID- 9862090 TI - Developmental hematopoiesis. AB - Hematopoiesis is a developmental process that evolves throughout the lifespan of an individual. Most work in the field has focused on events occurring in the adult bone marrow (BM). In the embryo, blood and endothelial cell generation begins very early after gastrulation, in defined intraembryonic mesodermic sites. Recent multidisciplinary studies, taking advantage of classic embryological and gene targeting technology in various species, have provided a new image of embryofetal lymphohemopoiesis, which includes the suggestion of developmental compartmentalization or waves. The first hematopoietic stem cells (HSC) migrate further and home in an ordered sequence of supporting microenvironments depending on scarcely known molecular requirements. These early hematopoietic progenitors show important differences in their cell biology and differentiation potentialities with respect to those present in adult stages; this fact, together with specific microenvironmental influences, define a process that diverges significantly from that occurring in the BM. Here, we update the latest developments in the field, the new understanding of lymphohemopoiesis in prenatal life, and the novel questions that this emerging paradigm is producing. PMID- 9862091 TI - The structure and function of gamma c-dependent cytokines and receptors: regulation of T lymphocyte development and homeostasis. AB - Five cytokines, IL-2, IL-4, IL-7, IL-9, and IL-15, form one group that is characterized by utilizing the common gamma chain (gamma c) as a receptor subunit. Examination of the phenotype of various cytokine or cytokine receptor "knockout" mice demonstrates that these cytokines are critical for normal lymphocyte development and subsequent functional activity of the peripheral immune compartment. This review summarizes the structural and functional properties of each of these five cytokines and their receptors, including the known redundant pathways for each cytokine or receptor. The contribution of these cytokines and receptors will then be considered in detail with respect to regulation of T lymphocyte development and homeostasis of the peripheral T cell compartment. PMID- 9862088 TI - Mood, cue and gender influences on motivation, craving and liking for alcohol in recreational drinkers. AB - The effects of exposure to an alcohol-related cue (drinking low-alcohol beer) and a musical depression/elation mood induction procedure, on craving, motivation and liking for alcohol, were studied in male and female recreational drinkers. Alcohol craving was assessed using the multidimensional desires for alcohol questionnaire (DAQ), motivation for alcohol was assessed by performance on a progressive ratio (PR) task reinforced with small volumes (25 ml) of low-alcohol beer, and liking for the reinforcers earned in the PR task was assessed using a visual analogue scale. Consumption of a half-pint of low-alcohol beer increased alcohol craving in male subjects but had no effect or decreased craving in female subjects. Subsequent induction of a depressed mood increased craving scores, relative to elated or neutral mood groups, but these effects were confined to abstinent (non-cued) subjects, both male and female. Performance on the PR task correlated significantly with one of the four factors of the DAQ, negative reinforcement, and was increased by induction of a depressed mood in abstinent female and cued male subjects. Reinforcer liking was unchanged following mood induction in male subjects, but decreased in both groups of female subjects. To summarize, the cue of drinking low-alcohol beer increased alcohol craving in men but not in women, and induction of a depressed mood increased alcohol craving and motivation, but also decreased alcohol liking. These effects were present to different extents in different cue/gender subgroups, and were partially independent. PMID- 9862087 TI - Multiple variable interval schedule behaviour in humans: effects of ethanol, mood, and reinforcer size on responding maintained by monetary reinforcement. AB - Ethanol is an effective reinforcer but, in common with other drugs of abuse, it may derive some of its reinforcing properties from the effects it has on behaviour maintained by other reinforcers. However, any assessment of ethanol's hypothesized effect on behaviour maintained by other reinforcers must take into account the fact that ethanol may have multiple mechanisms of action. In order to address this problem the experiments reported herein used a procedure based upon Herrnstein's Matching Law which allowed joint assessment of subjects' motor capacity and reinforcer sensitivity. The effect of ethanol (0, 0.3, and 0.6 g/kg) on motor capacity and reinforcer sensitivity was assessed by studying behaviour maintained by monetary reinforcement. In the first experiment the procedure was validated by showing that the behaviour of subjects was sensitive to changes in reinforcer value and in the second experiment 0.6 g/kg ethanol reduced motor capacity but did not affect reinforcer sensitivity. As a secondary hypothesis we also studied the effect of mood on reinforcer sensitivity and motor capacity. It was found that lower mood scores (lower hedonic tone) were associated with reduced reinforcer sensitivity and that male subjects showed higher motor capacity than females. However, there was also a mood by sex interaction, which indicated that higher motor capacity in males was only found in the presence of lower mood scores. The results are discussed in relation to the mechanisms of ethanol's dopaminergic effects, interactions between ethanol and other drugs of abuse, and the changes in reinforcer sensitivity which are thought to occur in depression. PMID- 9862089 TI - Alcohol choice and outcome expectancies in social drinkers. AB - Eighteen male social drinkers underwent four training sessions during which they ingested two colour-coded drinks (red or blue, balanced for drink type); one containing alcohol (aliquots of 0.1 g/kg) and the other placebo (aliquots of orangeade). Following the training sessions, subjects were presented with both drinks, and instructed to choose the drink they felt like consuming and to indicate their preference for their chosen drink over the other drink. In addition, they were instructed to consume the first drink but that all subsequent drinks (total of six drinks), offered at 10-min intervals, were optional. A number of trait characteristics were assessed including alcohol outcome expectancies, drinking habits and personality traits. The acute effects of alcohol on mood was also evaluated by comparing subjective ratings following alcohol and placebo during the training sessions. Of the 18 subjects, 12 chose alcohol at least once ('samplers'), whereas six never chose alcohol ('non samplers'). Over the three sessions, however, alcohol and placebo were chosen equally. When alcohol was chosen, subjects drank significantly more than when placebo was chosen, which may be consistent with a priming effect of drinking alcohol. The amount of alcohol drunk was seen to correlate with the alcohol expectancy factor 'sociability'. Subjective reports of feeling 'alert', 'clear headed', 'quick-witted', and 'attentive' all showed a main effect of choosing behaviour (i.e. 'samplers'/'non-samplers'). Further analysis indicated that this effect was due to 'samplers' reporting increased subjective ratings of these mood states following the ingestion of alcohol compared to 'non-samplers'. These increased subjective ratings were also positively correlated with the amount of alcohol consumed by the subjects during the choice procedure. No other relationships were found between the amount of alcohol consumed and any of the other state or trait measures. These data suggest that social drinkers who sample alcohol in a laboratory setting can be primed by alcohol to consume more. The results also indicated that the amount drunk was related to the degree to which subjects expected alcohol to increase sociability and to reports of subjective stimulant effects of alcohol (e.g., 'alert', 'clear-headed', 'quick-witted', and 'attentive'). PMID- 9862085 TI - Effects of marijuana on equilibrium, psychomotor performance, and simulated driving. AB - Delta-9-tetrahydrocannabinol (THC) is frequently found in the blood of drivers involved in automobile accidents, and marijuana use has been associated with impaired field sobriety test performance. The present study used a within-subject design to compare the effects of marijuana (0, 1.77, or 3.95% THC) on equilibrium and simulated driving. Ten marijuana users (seven men, three women) smoked one marijuana cigarette at the beginning of each session. Then 2 min later, they began a 60-min test battery that included subjective effects scales, a computerized test of body sway, a rapid judgment task and brake latency measurement in a driving simulator, critical flicker fusion (CFF), and a choice reaction time task (CRT). Self-report ratings of 'high' and 'drug potency' increased comparably following both active doses. The high, but not the low, dose significantly increased body sway. The high dose also marginally increased brake latency by a mean of 55 ms (P < 0.10), which is comparable to an increase in stopping distance of nearly 5 feet at 60 mph Judgment, CFF, and CRT scores did not differ across dose conditions. The equilibrium and brake latency data with 3.95% THC are similar to prior results in our laboratory in participants with breath alcohol concentrations near 0.05%. PMID- 9862086 TI - Mood and psychomotor performance effects of the first, but not of subsequent, cup of-coffee equivalent doses of caffeine consumed after overnight caffeine abstinence. AB - Moderate caffeine consumers (n = 64, mean caffeine intake 453 mg/day) were deprived of caffeine overnight and semi-randomly allocated to four treatment groups, designated PPP, CPP, CCP and CCC, where P is placebo and C is caffeine (1.2 mg/kg, giving an amount of caffeine similar to that consumed in a serving of ground coffee). Caffeine or placebo (i.e. no caffeine) were administered double blind in novel fruit juice drinks at 10:15, 11:30 and 13:00 h on the test day. Before (baseline), and 45 min after each of these times the participants completed a mood questionnaire and begun psychomotor performance tests lasting 25 min (1-min tapping task, and a long-duration simple reaction time task (SRT). Caffeine significantly increased energetic mood and improved psychomotor performance relative to placebo. Caffeine had particularly marked effects on SRT performance, ameliorating the slowing of performance with time on task and removing the post-lunch dip in performance. However, the three caffeine treatments, CPP, CCP and CCC, were equally effective. That is, mood and performance were improved to the same extent by one, two and three spaced doses (totalling 86, 172 and 258 mg) of caffeine. This result is consistent with previous findings indicating a flat dose-response relationship for the psychoactive effects of caffeine; and because of the adverse effects (e.g. fatigue) associated with overnight caffeine deprivation, it suggests that there is little net benefit to be gained from frequent caffeine use. At the very least, it appears that the psychostimulant effects of caffeine cannot on their own account for the typical pattern of consumption of caffeine-containing drinks. PMID- 9862082 TI - Effects of single oral administrations of haloperidol and d-amphetamine on prepulse inhibition of the acoustic startle reflex in healthy male volunteers. AB - The effects of acute administration of an indirect dopamine-agonist, d amphetamine, and a non-selective dopamine receptor antagonist, haloperidol, were investigated in normal male volunteers on habituation and prepulse inhibition (PPI) of the acoustic startle reflex in two experiments. In Experiment 1, 40 male non-smoker volunteers were tested for habituation and PPI (defined as percentage reduction of the pulse-alone amplitude; prepulses 9 dB above background) before and after double-blind administration of either 2 mg haloperidol or placebo. No influence of haloperidol was observed on either habituation or PPI of the startle reflex in this experiment. In Experiment 2, 60 male volunteers underwent startle testing before and after double-blind administration of a single oral dose of 5 mg haloperidol, 5 mg d-amphetamine or placebo. Habituation and PPI (prepulses 15 dB above background) for the placebo group did not differ significantly from that observed for the d-amphetamine or for the haloperidol group. However, in a subgroup of smoking subjects, both d-amphetamine and haloperidol reduced PPI as compared to that observed prior to drug administration. The implications of these findings in relation to animal pharmacological studies and observed sensorimotor gating deficits in schizophrenia are discussed. PMID- 9862097 TI - Ultrastructural changes in myocardium during mild hypothermic retrograde blood cardioplegia. AB - Uniformity of myocardial protection during retrograde blood cardioplegia is still a controversial area. We conducted a study on electron microscopic changes in the myocardium during mild hypothermic retrograde cardioplegia (31-32 degrees C) in 12 patients undergoing coronary artery bypass grafting. Biopsies for electron microscopy were taken from the right and left ventricular myocardium before and at the end of aortic cross-clamping and after 15 min reperfusion. The intercellular junctions, intracellular and extracellular oedema, mitochondria, capillaries, nuclei and myofibrils were analysed separately in each specimen, using a semiquantative method with scoring from 0 (unchanged) to 3 (severe changes), and the total scores were correlated with the severity of right and left coronary artery disease and with ischaemia time during aortic cross clamping. Mild to moderate ultrastructural changes occurred in the myocardium during the cardiopolegia, most typically myofibrillar injury and oedema. These changes increased during aortic cross-clamping and reperfusion, especially in the right ventricle. The total ultrastructural score for the right ventricle correlated negatively with the severity of right coronary artery disease at the end of cross-clamping. No such correlation was found in the left ventricle. Apart from one case of perioperative myocardial infarction, the clinical outcome was unproblematic. Myocardial structure thus was by and large well preserved during mild hypothermic retrograde blood cardioplegia, with the right ventricle seemingly somewhat less protected than the left. PMID- 9862094 TI - CD4+ T cells orchestrate both amplification and deletion of CD8+ T cells. AB - This review focuses on the role of CD4+ T cells in regulating immune responses, orchestrating both the amplification and deletion of immune cells, particularly CD8+ T cells. These two functions, which represent only an apparent contradiction, appear to be two faces of the same process of regulation. In fact, because the immune response, once activated, needs to be carefully controlled or switched off when the antigenic stimulus is eliminated, the immune system has developed several strategies either to regulate clonal amplification or to avoid useless expansion of activated cells. In particular, we have reported many data demonstrating that CD4+ T cells may be indicated as the regulatory element in the activation as well as the deletion of CD8+ T cells. New data are also reported on the ability of anergic CD4+ T cells to suppress CD8+ T-cell activation through induction of apoptosis, and on the need for CD8+ T cells for antigen recognition in inducing cell death in CD4+ T cells. Moreover, the central role of CD4+ T cells in the maintenance of peripheral tolerance has been widely described. PMID- 9862095 TI - Role of echocardiography in systemic arterial embolism. A review with recommendations. AB - The ability of echocardiography to diagnose sources of embolism and the role of the examination in the prediction of thromboembolism are reviewed. In addition, the yield of transthoracic (TTE) and transoesophageal echocardiography (TEE) is analysed in patients with suspected embolism and guidelines are proposed for performing echocardiography in this setting. In general, echocardiography is reliable for diagnosing sources of embolism and this applies in particular to TEE in the case of atrial, valvular, and aortic abnormalities. However, the method is useful for predicting embolism in a few cases only. There is a substantial risk in the event of mobile or protruding thrombi, but screening for these and other markers of thromboembolism seems to be unproductive in most groups of risk patients. Yet, in the presence of atrial fibrillation, echocardiography may be helpful in defining patients with an otherwise normal heart and low risk of embolism--and in defining the relatively rare patient with a clinically low-risk profile but moderate-to-severe left ventricular systolic dysfunction and a high risk of embolism. TEE-guided conversion of atrial fibrillation without weeks of preceding anticoagulation may prove useful, after further investigation. The risk of embolism in relation to the size and mobility of valvular vegetations has remained controversial. In patients with suspected recent embolism, TTE results in less than 5% new therapeutic consequences. In those with a normal TTE, the yield of TEE seems to be equally low. We therefore recommend a selective strategy: TTE and TEE can be omitted when a cardiac source of embolism appears from the clinical setting and in most patients with an obvious predisposition to cerebrovascular disease. However, in the latter cases TTE should be performed if indicated by the clinical situation, e.g. in the presence of fever and murmur. TTE is also recommended when there are no obvious markers of primary vascular disease. To preclude very rare sources of embolism (e.g. atrial thrombi despite sinus rhythm), supplementary TEE is recommended in younger patients in whom primary vascular disease is very unlikely. The diagnosis by TEE of common conditions such as atrial septal aneurysms and patent foramen ovale cannot, however, be taken as proof of the mechanism of a systemic arterial occlusive event; thus it is difficult to change therapy on the basis of such diagnoses. PMID- 9862096 TI - Angiographic changes in saphenous vein grafts and atherosclerosis risk factors. A 5-year study with serial measurements of serum lipids and lipoproteins. AB - The association between cardiovascular risk factors and stenosis or occlusion of saphenous vein grafts was analysed in a prospective 5-year study of 176 unselected patients with coronary artery bypass grafting (CABG). Methods included serial measurements of serum lipids and lipoproteins, determination of apolipoprotein E phenotype, lipoprotein (a) levels 5 years postoperatively, and subcutaneous fat biopsy to determine the fatty acid composition before and one year after CABG. Graft angiography with quantitative analysis of angiograms was performed at the end of follow-up. A coronary artery with diameter < or = 1.5 mm was associated with occlusion of vein grafts (p < 0.01). The mean levels of serum lipids and lipoproteins, other traditional risk factors for atherosclerosis, and subcutaneous fatty acid composition were similar in patients with and without graft occlusion, and similar when the maximum diameter of non-occluded grafts was < 50% vs > or = 50%, and < 25% vs > or = 25%. High lipoprotein (a) concentration tended to be associated with obstructive changes in vein grafts. Our data indicate that, because lipids, lipoproteins and other traditional cardiovascular risk factors do not predict occlusion or stenosis of saphenous vein grafts five years after CABG, it is not currently possible to predict directly from the levels of these risk factors which patients are likely to benefit from pharmacological or other interventions. PMID- 9862098 TI - Plasma N-terminal atrial natriuretic peptide predicts hospitalization in patients with heart failure. AB - The plasma concentration of N-terminal atrial natriuretic peptide (N-ANP) has been shown to be predictive of both clinical status and survival in patients with heart failure. In this analysis the relationship between N-ANP, morbidity and hospitalization time was evaluated in 417 patients with stable, congestive heart failure recruited from an active, outpatient heart failure registry. Hospital admissions along with the duration of stay occurring after the initial N-ANP sampling during the period of data collection were recorded. A total of 755 admissions occurred, accounting for 7917 days' hospitalization. Relative hospitalization times (in-hospital days/observation period) per N-ANP quartiles I IV were: 1.2 (+/- 2.7)%, 5.5 (+/- 12.2)%, 10.0 (+/- 21.5)% and 20.8 (+/- 34.3)%, respectively. Although N-ANP levels were correlated with age (r = 0.234, p < 0.0001), division by age quartiles did not significantly predict relative hospitalization times. These data indicate that the degree of cardiac endocrine activation and subsequent N-ANP release is related to morbidity in patients with heart failure and that moderate elevation in N-ANP levels is associated with a substantially increased hospitalization time. N-ANP sampling should be of value as a supplement to clinical evaluation in the assessment of the individual patient with this common syndrome. PMID- 9862100 TI - Unusual course of right atrial myxoma, masked by acute abdominal pain, and complicated by pulmonary embolus. AB - In a 17-year-old youth, exploratory laparotomy for acute abdominal pain was complicated by circulatory arrest related to pulmonary embolism. Echocardiography after resuscitation revealed a hitherto "silent" right atrial myxoma, fragmentation of which had blocked the right atrioventricular ostium, causing the pain. Operation was successful. As only a few similar cases have been reported, pulmonary embolism due to fragmentation of a right atrial myxoma may account for some unexplained sudden deaths beyond medical help. PMID- 9862101 TI - The effects of tissue expansion on skin lymph flow and lymphatics: an experimental study in rabbits. AB - Histomorphological, physiological, and biomechanical changes of the skin and tissue that are being expanded have been extensively studied in the past. The purpose of this study was to investigate whether the skin lymph flow and lymphatics may be influenced during tissue expansion, and also to study different pattern of vascular supply of skin on lymph flow during tissue expansion. The skin lymph flow was quantitatively assessed by 99mTc-dextran lymphoscintigraphy, and the structure of dermal lymphatics was evaluated by histological examination in 12 rabbits. Lymphoscintigraphic results showed that lymph flow is significantly reduced both in expanded and non-expanded (sham-operated) skin. Histologically, we saw widespread lymphatic distension in both expanded and non expanded skin. There were no quantitative differences in the number of lymphatic vessels compared with control skin. We did not see any lymphatic capillaries in the subdermal capsule of either expanded or non-expanded skin. These results show that lymphostasis has an obstructive (mechanical) aetiology, and the tissue expander itself reduces the lymph flow regardless of expansion, and interferes with the formation of new lymphatic vessels. PMID- 9862084 TI - The effects of memantine on the subjective, reinforcing and cardiovascular effects of cocaine in humans. AB - Eight male frequent cocaine smokers participated in a 44- to 47-day inpatient and outpatient study to assess the effects of the noncompetitive N-methyl-D-aspartate (NMDA) antagonist, memantine, on cocaine self-administration, subjective effects, and psychomotor performance. Participants were maintained on memantine (0 and 20 mg daily) for 7-10 days prior to laboratory testing, using a double-blind crossover design. Under each medication condition, participants smoked four doses of cocaine base (0, 12, 25 and 50 mg), and were subsequently given five opportunities, 14 min apart, to self-administer that dose of cocaine or receive a merchandise voucher ($5.00). Each cocaine dose was tested twice under each medication condition, and the order of medication condition and cocaine dose was systematically varied. Vital signs were recorded every 2 min, and subjective effects were assessed at baseline and after each cocaine or voucher delivery. In addition, psychomotor performance was assessed before and after each self administration session. Memantine maintenance was not associated with changes in psychomotor performance or the number of cocaine doses chosen each session. Memantine maintenance was, however, associated with significant increases in some subjective effects of cocaine, including ratings of 'good drug effect', 'high', 'potency', 'quality', and street value. These data suggest that NMDA antagonists may have limited usefulness as treatment medications for cocaine abuse. PMID- 9862093 TI - Immune deficits induced by strenuous exertion under adverse environmental conditions: manifestations and countermeasures. AB - A brief description is given of the various laboratory and clinical manifestations of immune suppression that arise when strenuous exertion must be carried out in the face of a negative energy balance, shifts of circadian rhythm, sleep deprivation, psychological stressors, and exposure to hostile environments (extremes of heat or cold, high or low ambient pressures, and hyper- or hypo gravity conditions). From the operational viewpoint, immune suppression could impair both physical and mental performance by increasing susceptibility to opportunistic microorganisms. It is also likely to increase susceptibility to sepsis following trauma or extensive burns, and has occasionally predisposed to fatal myocarditis. The effects of such challenges are complex, in part because of interactions between the various stressors. It is thus important to investigate the impact and to devise appropriate countermeasures with the full physical and intellectual resources of a defense environmental research laboratory. Existing knowledge of the topic is reviewed, and suggestions are made for research that may lead to new and more effective countermeasures. PMID- 9862102 TI - Histodifferentiation of hair follicles in grafting of cell aggregates obtained by rotation culture of embryonic rat skin. AB - We have previously reported reconstruction of hair follicles from a single cell suspension of rat fetal upper lip by a two-step culture method consisting of rotation and flotation cultures. Rotation sorted out the cells and flotation facilitated histodifferentiation. In the present study, we added grafting procedures to the previous method to see whether cell aggregates obtained this way were graftable, and whether the grafting promoted histodifferentiation. The aggregates before and after flotation were grafted, and differentiation of hair follicles comparable to those in vivo was confirmed 10 days after grafting. There was no difference in the degree of differentiation between the two kinds of grafts. The grafting procedure therefore resulted in an appreciable increase in histodifferentiation even when aggregates obtained after flotation were grafted. PMID- 9862105 TI - Immobilised heparin accelerates the healing of human wounds in vivo. AB - Locally produced growth factors are of great importance in wound healing in human skin. Wound fluid from chronic wounds contains low concentrations of growth factors possibly because of rapid degradation as a result of the high concentration of proteases. Many growth factors involved in wound healing bind to heparin and are thereby stabilised and activated. We have recently shown that heparin in combination with chitosan stimulates re-epithelialisation in an in vitro model of human wound healing. In the present study we investigated the effects of a chitosan-heparin membrane on wound healing in 10 split-thickness graft donor sites in human skin. The chitosan-heparin membrane stimulated healing of the donor sites both when judged macroscopically in a blinded fashion and when biopsy specimens from the treated and untreated parts of the wound were investigated microscopically. We hypothesise that the beneficial effects of the chitosan-heparin membrane result from slow release of heparin into the wound area which protects locally produced growth factors. The result is increased stabilisation and concentration of growth factors in the wound area, which stimulate healing. We believe that these results may be important in the treatment of wounds that are reluctant to heal. PMID- 9862103 TI - E-selectin and L-selectin blockade in pure skin flaps exposed to ischaemia and reperfusion injury. AB - The inflammatory recruitment of leucocytes is a main cause of tissue damage in ischaemia/reperfusion (I/R) injury. Under appropriate flow conditions, E-selectin and L-selectin participate in the initial deceleration of neutrophils (PMNs) on inflamed endothelial cells before transmigration of PMNs into the surrounding tissue. Previous work from our lab showed increased survival of I/R injured myocutaneous flaps after treatment with anti-E/L-selectin. In this study, we have evaluated a combined antibody to E-selectin and L-selectin (EL-246) in porcine pure skin flaps exposed to I/R injury. Buttock skin flaps were exposed to eight hours of ischaemia and 20 hours of reperfusion. EL-246 or saline was given intra arterially into the flaps. Estimated surviving area was not improved in the treated group. The lack of effect of EL-246 supports our suspicion that different mechanisms are involved in I/R injury in myocutaneous flaps compared with pure skin flaps. As a certain shear stress must be present for the selectins to exert their effect, a possible explanation for the diverse results in muscle and skin might be different reflow patterns. PMID- 9862099 TI - Improvement in long-term prognosis of elderly patients with acute myocardial infarction after the introduction of intravenous thrombolytic therapy. AB - Survival rate from a "thrombolytic" period of 351 patients above 66 years of age with acute myocardial infarction (AMI) was compared with that of 289 patients from a "prethrombolytic" period. The two groups were comparable regarding sex, age, previous AMI, cerebrovascular events, morbidity and mortality during admission. Survival rates after four years were 45.0% in the "thrombolytic" group and 38.4% in the "prethrombolytic" group (p = 0.047, log rank test). Using the Cox proportional hazard analysis, thrombolytic therapy was shown to be an independent prognostic predictor in "the thrombolytic population" with a relative risk of death from day 30 to end of follow-up of 0.4 (95% confidence interval 0.2 0.8). No interaction was found between age and thrombolysis. Although only one fifth of the patients with AMI were eligible for thrombolysis, this treatment may have contributed to the improved long-term survival. PMID- 9862104 TI - Improvement in nerve regeneration by monoclonal antibodies to ICAM-1 and LFA-1 in allogeneic mice. AB - We examined whether giving monoclonal antibodies (MoAb) to intercellular adhesion molecule 1 (ICAM-1) and leucocyte function associated antigen (LFA-1), which have important roles in the initial stage of rejection after allografts, can improve nerve regeneration in allogeneic mice. Fresh sciatic nerves were grafted using BALB/c mice as donors and C3H/He mice as recipients. Nerve regeneration at six weeks was significantly better in the mice given MoAbs at one and five days than in those given none (n = 5 in each group), although nerve regeneration even in the five-day group was significantly inferior to that in the syngeneic nerve graft group (n = 5). The survival time of the nerve donor skin graft at 12 weeks was not prolonged by treatment with MoAbs, indicating a failure to induce immunological tolerance. However, at 10 months after nerve grafting there were fewer Mac-1, Lyt-1, and Thy-1 positive cells in the five-day group and they showed less immunoreactivity than the untreated group. We conclude that giving MoAbs could effectively improve nerve regeneration in grafted allogeneic nerve segments, although it did not induce immunological tolerance. PMID- 9862106 TI - Millard repair of unilateral isolated cleft lip: a 25-year follow-up. AB - Twenty-five patients with isolated unilateral cleft lip took part in a follow-up study at a mean age of 28.6 years. All had had a primary, Millard lip repair, at a mean age of 4.6 months. In 20 patients, at least one secondary correction had been undertaken during adolescence. The overall long-term outcome was thought to be good, leaving a fairly inconspicuous fine lip scar and acceptable nose configuration. In half the patients, however, the lip was slightly elongated and the nostrils were still asymmetrical. These findings concurred with the patients' subjective assessments, which showed that appearance of the lip and nose were rated good by 20 (80%) and 16 (64%), respectively. The findings of this study provide a baseline for future evaluation of the results achieved with lip closure by Johanson's technique. PMID- 9862107 TI - The effect of muscle repair on postoperative facial skeletal growth in children with bilateral cleft lip and palate. AB - The effect of orbicularis muscle repair on postoperative facial skeletal growth in bilateral cleft lip and palate patients was studied by analysis of cephalometric radiographs and dental casts. Sixty-two patients operated on between 1961-1989 were selected for the study. They were divided into three groups, group 1a (muscle repair; n = 12), group 1b (failed attempt at muscle repair; n = 5), and group 2 (no attempt at muscle repair; n = 45). Comparison of the morphological measurements among these three groups showed that there was a trend towards crossbite in the muscle repair group, but this difference was not significant. Mechanisms by which muscle repair might influence maxillofacial skeletal growth include the possibility that the area around the nasal septum might be the growth centre. The choice of operative technique in bilateral cleft lip and palate should be important. PMID- 9862108 TI - Comparison of the effect of a collagen dressing and a polyurethane dressing on the healing of split thickness skin graft (STSG) donor sites. AB - Recent advances in the resurfacing of burn wounds with dermal equivalents and collagen preparations have shown the efficacy of collagen. To investigate the benefits (if any), standardised split skin donor areas were chosen to compare the influence of collagen on re-epithelialisation. A bovine collagen preparation consisting of type-I collagen was prospectively compared with polyurethane film dressing in a study of 20 split thickness skin graft donor sites. The rates of epithelialisation, the discomfort experienced by the patients and the convenience of the dressings were assessed. The median time from operation to the observation of complete healing was 7.5 (+/- 2.5) days for the donor sites dressed with the collagen membrane and 12.5 (+/- 3.4) days for the the donor areas dressed with a polyurethane film (p < 0.001). The discomfort experienced by the two groups of patients was significantly less after wound coverage with collagen (p < 0.005). Haematomas or seromas that required repeated aspiration was seen under the polyurethane film dressing. The collagen dressing was more expensive than the polyurethane film, but improved wound healing compared with the polyurethane dressings. PMID- 9862109 TI - Early dermabrasion of deep dermal burns with sandpaper. Case reports. AB - Deep dermal burns are initially difficult to evaluate, and they sometimes heal spontaneously. We present our experience of dermabrasion with sandpaper in four patients. It is a useful alternative to early excision of the scar. Skin grafts are not always required and the aesthetic results are excellent. Dermabrasion should be considered routinely for all deep dermal burns and particularly for facial burns and those caused by scalds. PMID- 9862110 TI - Pseudoxanthoma elasticum. Case report. AB - Pseudoxanthoma elasticum is an uncommon disorder of elastin with characteristic skin laxity. Rhytidectomy of the excessive redundant skin is necessary for cosmetic improvement, and reports of surgical management are rare. We report a case of pseudoxanthoma elasticum with involvement of neck, axillae, breasts, abdomen, thighs, and perineum, together with angioid streaking of fundi. Rhytidectomy of the lax cervical skin was undertaken with satisfactory results after a 13 month follow-up. We have reviewed other published reports to obtain a better understanding of the genetics, histopathology, vascular and ophthalmological manifestations, and also the surgical implications of pseudoxanthoma elasticum. PMID- 9862111 TI - Common peroneal nerve palsy caused by a ganglion. Case report. AB - A common peroneal nerve palsy caused by a ganglion cyst is a rare entity. A 48 year old man was referred with a six year history of intermittent pain that had resulted in a complete palsy of the common peroneal nerve. A magnetic resonance (MR) scan showed compression of the nerve by a ganglion, which was excised and the nerve was released. Three months later the lost muscle function had returned completely. PMID- 9862113 TI - Fracture of the metacarpal head with 180 degrees longitudinal rotation. Case report. AB - A 61-year old man fell and sustained an intracapital fracture of the 3rd metacarpal bone with an unusual dislocation. Primary radiograph showed 180 degrees longitudinal rotation of the distal fragment. To reduce the fracture the metacarpophalangeal joint was opened and the fracture stabilised. The fracture healed after four weeks, and there was no sign of necrosis at the three month follow up. PMID- 9862112 TI - An unusual case of deep penetrating dermatofibroma of the lower leg. Case report. AB - Histological examination of a punch biopsy specimen of a slowly growing, irregular dermal tumour from the right lower leg of a 32-year-old woman, which had been diagnosed clinically as a dermatofibrosarcoma protuberans, showed it to be a deep penetrating dermatofibroma. The diagnosis was established by characteristic histological and immunohistochemical criteria. This allowed sequential excision in two sessions with primary closure and an optimal cosmetic result. PMID- 9862114 TI - Distortion of the radial artery by a mucinous cyst. Case reports. AB - Three cases of mucinous cyst situated close to the radial artery are reported. The patients complained of pain, a throbbing mass, or both at the wrist. Colour Doppler sonography showed distortion of the radial artery by the cyst in all three patients. In one patient the cyst was connected to a synovial sac by a pedicle, in another it was adherent to the radial artery but was identified histopathologically as a simple ganglion, and in the last patient a branch of the radial artery was involved in the cyst, which was identified histopathologically as an adventitial cyst. Mucinous cysts enlarge when subjected to mechanical stress. Excision is recommended for cysts that distort the radial artery. PMID- 9862117 TI - [Purchase and purchase examination of psittacines]. PMID- 9862115 TI - [Dental problems in rabbits and rodents. I. Anatomy, physiology, symptomatology, diagnosis]. AB - Dental and oral problems in rodents and rabbits are common in veterinary practice. Anatomy and physiology of the different dentition, the common symptoms and the clinical diagnosis are discussed in the first part of this article. In the next article treatment and prevention are described. PMID- 9862118 TI - [Interview with Professors Mossel and Struijk. Consumer protection is the job of veterinarians]. PMID- 9862119 TI - [Report of the mastitis panel, May, 1998. "Special microbes"]. PMID- 9862120 TI - Microbial genomics. PMID- 9862116 TI - [Dental problems in rabbits and rodents. II. Dental treatment]. AB - Anatomy and physiology of the different dentition, the common symptoms and the clinical diagnosis are discussed in the first part of the article on dental and oral problems in rodents and rabbits. In this article (the second part) prognosis, treatment plan, treatment and prevention are described. PMID- 9862121 TI - Indigo: a World-Wide-Web review of genomes and gene functions. AB - The present article describes a genome database reviewing gene-related knowledge of two model bacteria, Bacillus subtilis and Escherichia coli. The database, Indigo, is open through the World-Wide Web (http://indigo.genetique.uvsq.fr). The concept used for organising the data, the concept of neighbourhood, allows one to explore the database content in an efficient although somewhat unusual way. Here, genes are related to each other by a variety of neighbourhoods, including proximity in the chromosome, phylogenetic kinship, participation in a common metabolic pathway, common presence in an article of the literature, or similar use of the genetic code. Several examples illustrate how this concept of neighbourhood permits one to review the available knowledge about a given gene or gene family, and elaborate unexpected, but revealing, analyses about gene functions. PMID- 9862124 TI - Hydropathy profile alignment: a tool to search for structural homologues of membrane proteins. AB - Hydropathy profile alignment is introduced as a tool in functional genomics. The architecture of membrane proteins is reflected in the hydropathy profile of the amino acid sequence. Both secondary and tertiary structural elements determine the profile which provides enough sensitivity to detect evolutionary links between membrane proteins that are based on structural rather than sequence similarities. Since structure is better conserved than amino acid sequence, the hydropathy profile can detect more distant evolutionary relationships than can be detected by the primary structure. The technique is demonstrated by two approaches in the analysis of a subset of membrane proteins coded on the Escherichia coli and Bacillus subtilis genomes. The subset includes secondary transporters of the 12 helix type. In the first approach, the hydropathy profiles of proteins for which no function is known are aligned with the profiles of all other proteins in the subset to search for structural paralogues with known function. In the second approach, family hydropathy profiles of 8 defined families of secondary transporters that fall into 4 different structural classes (SC-ST1-4) are used to screen the membrane protein set for members of the structural classes. The analysis reveals that over 100 membrane proteins on each genome fall in only two structural classes. The largest structural class, SC-ST1, correlates largely with the Major Facilitator Superfamily defined before, but the number of families within the class has increased up to 57. The second large structural class, SC-ST2 contains secondary transporters for amino acids and amines and consists of 12 families. PMID- 9862123 TI - Genome evolution within the alpha Proteobacteria: why do some bacteria not possess plasmids and others exhibit more than one different chromosome? AB - Animal intracellular Proteobacteria of the alpha subclass without plasmids and containing one or more chromosomes are phylogenetically entwined with opportunistic, plant-associated, chemoautotrophic and photosynthetic alpha Proteobacteria possessing one or more chromosomes and plasmids. Local variations in open environments, such as soil, water, manure, gut systems and the external surfaces of plants and animals, may have selected alpha Proteobacteria with extensive metabolic alternatives, broad genetic diversity, and more flexible and larger genomes with ability for horizontal gene flux. On the contrary, the constant and isolated animal cellular milieu selected heterotrophic alpha Proteobacteria with smaller genomes without plasmids and reduced genetic diversity as compared to their plant-associated and phototrophic relatives. The characteristics and genome sizes in the extant species suggest that a second chromosome could have evolved from megaplasmids which acquired housekeeping genes. Consequently, the genomes of the animal cell-associated Proteobacteria evolved through reductions of the larger genomes of chemoautotrophic ancestors and became rich in adenosine and thymidine, as compared to the genomes of their ancestors. Genome organisation and phylogenetic ancestor-descendent relationships between extant bacteria of closely related genera and within the same monophyletic genus and species suggest that some strains have undergone transition from two chromosomes to a single replicon. It is proposed that as long as the essential information is correctly expressed, the presence of one or more chromosomes within the same genus or species is the result of contingency. Genetic drift in clonal bacteria, such as animal cell-associated alpha Proteobacteria, would depend almost exclusively on mutation and internal genetic rearrangement processes. Alternatively, genomic variations in reticulate bacteria, such as many intestinal and plant cell-associated Proteobacteria, will depend not only on these processes, but also on their genetic interactions with other bacterial strains. Common pathogenic domains necessary for the invasion and survival in association with cells have been preserved in the chromosomes of the animal and plant-associated alpha Proteobacteria. These pathogenic domains have been maintained by vertical inherence, extensively ameliorated to match the chromosome G + C content and evolved within chromosomes of alpha Proteobacteria. PMID- 9862125 TI - The genome of Treponema pallidum: new light on the agent of syphilis. AB - Treponema pallidum subsp, pallidum, the causative agent of the sexually transmitted disease syphilis, is a fastidious, microaerophilic obligate parasite of humans. This bacterium is one of the few prominent infectious agents that has not been cultured continuously in vitro and consequently relatively little is known about its virulence mechanisms at the molecular level. T. pallidum therefore represented an attractive candidate for genomic sequencing. The complete genome sequence of T. pallidum has now been completed and comprises 1,138,006 base pairs containing 1041 predicted protein coding sequences. An important goal of this project is to identify possible virulence factors. Analysis of the genome indicates a number of potential virulence factors including a family of 12 proteins related to the Msp protein of Treponema denticola, a number of putative hemolysins, as well as several other classes of proteins of interest. The results of this analysis are reviewed in this article and indicate the value of whole genome sequences for rapidly advancing knowledge of infectious agents. PMID- 9862122 TI - The serine, threonine, and/or tyrosine-specific protein kinases and protein phosphatases of prokaryotic organisms: a family portrait. AB - Inspection of the genomes for the bacteria Bacillus subtilis 168, Borrelia burgdorferi B31, Escherichia coli K-12, Haemophilus influenzae KW20, Helicobacter pylori 26695, Mycoplasma genitalium G-37, and Synechocystis sp PCC 6803 and for the archaeons Archaeoglobus fulgidus VC-16 DSM4304, Methanobacterium thermoautotrophicum delta H, and Methanococcus jannaschii DSM2661 revealed that each contains at least one ORF whose predicted product displays sequence features characteristic of eukaryote-like protein-serine/threonine/tyrosine kinases and protein-serine/threonine/tyrosine phosphatases. Orthologs for all four major protein phosphatase families (PPP, PPM, conventional PTP, and low molecular weight PTP) were present in the bacteria surveyed, but not all strains contained all types. The three archaeons surveyed lacked recognizable homologs of the PPM family of eukaryotic protein-serine/threonine phosphatases; and only two prokaryotes were found to contain ORFs for potential phosphatases from all four major families. Intriguingly, our searches revealed a potential ancestral link between the catalytic subunits of microbial arsenate reductases and the protein tyrosine phosphatases; they share similar ligands (arsenate versus phosphate) and features of their catalytic mechanism (formation of arseno-versus phospho cysteinyl intermediates). It appears that all prokaryotic organisms, at one time, contained the genetic information necessary to construct protein phosphorylation dephosphorylation networks that target serine, threonine, and/or tyrosine residues on proteins. However, the potential for functional redundancy among the four protein phosphatase families has led many prokaryotic organisms to discard one, two, or three of the four. PMID- 9862127 TI - Microdrop screening: a rapid method to optimize solvent conditions for NMR spectroscopy of proteins. AB - Determining appropriate solvent conditions is a crucial first step for carrying out NMR spectroscopy of proteins, but rapid and efficient methods for doing so are currently lacking. Microdrop screening examines a large number of different solvent conditions using very small amounts of protein and minimal labor. Starting from one initial buffer condition, small aliquots of protein solution are combined with an array of solutions in which concentration, pH, buffer type, and added stabilizers are systematically varied. The protein concentration of each microliter-sized test drop ('microdrop') is gradually changed using vapor diffusion, and the solubility of the protein is determined by visual examination. A variety of analytical techniques may be applied to the contents of the microdrops to monitor enzymatic activity, aggregation, ligand binding, and protein folding. PMID- 9862126 TI - Propagation of experimental uncertainties using the Lipari-Szabo model-free analysis of protein dynamics. AB - In this paper we make use of the graphical procedure previously described [Jin, D. et al. (1997) J. Am. Chem. Soc., 119, 6923-6924] to analyze NMR relaxation data using the Lipari-Szabo model-free formalism. The graphical approach is advantageous in that it allows the direct visualization of the experimental uncertainties in the motional parameter space. Some general 'rules' describing the relationship between the precision of the relaxation measurements and the precision of the model-free parameters and how this relationship changes with the overall tumbling time (tau m) are summarized. The effect of the precision in the relaxation measurements on the detection of internal motions not close to the extreme narrowing limit is analyzed. We also show that multiple timescale internal motions may be obscured by experimental uncertainty, and that the collection of relaxation data at very high field strength can improve the ability to detect such deviations from the simple Lipari-Szabo model. PMID- 9862129 TI - High pressure NMR study of a small protein, gurmarin. AB - The effect of pressure on the structure of gurmarin, a globular, 35-residue protein from Gymnema sylvestre, was studied in aqueous environment (95% 1H2O/5% 2H2O, pH 2.0) with an on-line variable pressure NMR system operating at 750 MHz. Two-dimensional TOCSY and NOESY spectra were measured as functions of pressure between 1 and 2000 bar at 40 degrees C. Practically all the proton signals of gurmarin underwent some shifts with pressure, showing that the entire protein structure responds to, and is altered by, pressure. Most amide protons showed different degrees of low field shifts with pressure, namely 0-0.2 ppm with an average of 0.051 ppm at 2000 bar, showing that they are involved in hydrogen bonding and that these hydrogen bonds are shortened by pressure by different degrees. The tendency was also confirmed that the chemical shifts of the amide protons exposed to the solvent (water) are more sensitive to pressure than those internally hydrogen bonded with carbonyls. The pressure-induced shifts of the H alpha signals of the residues in the beta-sheet showed a negative correlation with the 'folding' shifts (difference between the shift at 1 bar and that of a random coil), suggesting that the main-chain torsion angles of the beta-sheet are slightly altered by pressure. Significant pressure-induced shifts were also observed for the side-chain protons (but no larger than 10% of the 'folding' shifts), demonstrating that the tertiary structure of gurmarin is also affected by pressure. Finally, the linearity of the pressure-induced shifts suggest that the compressibility of gurmarin is invariant in the pressure range between 1 and 2000 bar. PMID- 9862128 TI - The NMR solution structure and characterization of pH dependent chemical shifts of the beta-elicitin, cryptogein. AB - The NMR structure of the 98 residue beta-elicitin, cryptogein, which induces a defence response in tobacco, was determined using 15N and 13C/15N labelled protein samples. In aqueous solution conditions in the millimolar range, the protein forms a discrete homodimer where the N-terminal helices of each monomer form an interface. The structure was calculated with 1047 intrasubunit and 40 intersubunit NOE derived distance constraints and 236 dihedral angle constraints for each subunit using the molecular dynamics program DYANA. The twenty best conformers were energy-minimized in OPAL to give a root-mean-square deviation to the mean structure of 0.82 A for the backbone atoms and 1.03 A for all heavy atoms. The monomeric structure is nearly identical to the recently derived X-ray crystal structure (backbone rmsd 0.86 A for residues 2 to 97) and shows five helices, a two stranded antiparallel beta-sheet and an omega-loop. Using 1H,15N HSQC spectroscopy the pKa of the N- and C-termini, Tyr12, Asp21, Asp30, Asp72, and Tyr85 were determined and support the proposal of several stabilizing ionic interactions including a salt bridge between Asp21 and Lys62. The hydroxyl hydrogens of Tyr33 and Ser78 are clearly observed indicating that these residues are buried and hydrogen bonded. Two other tyrosines, Tyr47 and Tyr87, show pKa's > 12, however, there is no indication that their hydroxyls are hydrogen bonded. Calculations of theoretical pKa's show general agreement with the experimentally determined values and are similar for both the crystal and solution structures. PMID- 9862130 TI - Determination of the equilibrium micelle-inserting position of the fusion peptide of gp41 of human immunodeficiency virus type 1 at amino acid resolution by exchange broadening of amide proton resonances. AB - The exchange broadening of backbone amide proton resonances of a 23-mer fusion peptide of the transmembrane subunit of HIV-1 envelope glycoprotein gp41, gp41 FP, was investigated at pH 5 and 7 at room temperature in perdeuterated sodium dodecyl sulfate (SDS) micellar solution. Comparison of resonance peaks for these pHs revealed an insignificant change in exchange rate between pH 5 and 7 for amide protons of residues 4 through 14, while the exchange rate increase at neutral pH was more prominent for amide protons of the remaining residues, with peaks from some protons becoming undetectable. The relative insensitivity to pH of the exchange for the amide protons of residues 4 through 14 is attributable to the drastic reduction in [OH-] in the micellar interior, leading to a decreased exchange rate. The A15-G16 segment represents a transition between these two regimes. The data are thus consistent with the notion that the peptide inserts into the hydrophobic core of a membrane-like structure and the A15-G16 dipeptide is located at the micellar-aqueous boundary. PMID- 9862134 TI - Assignments of 1H and 13C resonances in the complex of palmitate and a non specific lipid transfer protein (ns-LTP) isolated from barley seeds. PMID- 9862136 TI - Iridoid glycoside biosynthesis in Penstemon secundiflorus. Another H-5, H-9 trans iridoid glycoside. AB - Isolation and characterization of the new iridoid 10-hydroxy-(5 alpha H)-6 epidihydrocornin from Penstemon secundiflorus (Scrophulariaceae) is described. In biosynthetic experiments, deoxyloganic acid was incorporated into the trans-fused iridoid glycosides (5 alpha H)-6-epidihydrocornin and 10-hydroxy-(5 alpha H)-6 epidihydrocornin in P. secundiflorus. Formation of the trans-fused compounds is therefore a late event in the biosynthesis and does not occur during iridoid formation by cyclization of the open chain monoterpene precursor. In the same plant, 8-epideoxyloganic acid was not incorporated into the trans-iridoids. Deoxyloganic acid was also incorporated into 10-hydroxyhastatoside (which bears an 8 beta-methyl group), while 8-epideoxyloganic acid was incorporated into penstemoside (with an 8 alpha-methyl group). Thus, iridoid biosynthetic pathways leading from both deoxyloganic acid and 8-epideoxyloganic acid were found in the same plant. PMID- 9862138 TI - Amino acid sequence of ferredoxin from Physalis alkekengi var. francheti. AB - The complete amino acid sequence of [2Fe-2S] ferredoxin from Physalis alkekengi var. francheti has been determined by automated Edman degradation of the entire Cm-protein and of the peptides obtained by trypsin and endoproteinase Asp-N digestions. This ferredoxin exhibited ten, ten, and nine differences respectively in the amino acid sequence, when compared with the ferredoxins of Datura stramonium, D. metel, and D. arborea, but 21-28 differences for other angiosperms, and 34-37 differences for fern and horsetails. These results are in harmony with the taxonomic position for these plants. PMID- 9862132 TI - 1H and 15N resonance assignment of the calcium-bound form of the Nereis diversicolor sarcoplasmic Ca(2+)-binding protein. PMID- 9862135 TI - Purification and characterization of cannabichromenic acid synthase from Cannabis sativa. AB - Cannabichromenic acid synthase was purified to apparent homogeneity by sequential column chromatography including DEAE-cellulose, phenyl-Sepharose CL-4B, and hydroxylapatite. The enzyme catalysed the oxidocyclization of cannabigerolic acid and cannabinerolic acid to cannabichromenic acid. The K(m) values for both substrates were in the same order of magnitude although the Vmax value for the former was higher than that for the latter. These results suggested that cannabichromenic acid is predominantly formed from cannabigerolic acid rather than cannabinerolic acid. The enzyme required neither molecular oxygen nor hydrogen peroxide, indicating that the cannabichromenic acid synthase reaction proceeds through direct dehydrogenation without hydroxylation. PMID- 9862133 TI - 1H and 15N resonance assignment of neural cell adhesion molecule module-2. PMID- 9862140 TI - Anti-HIV-1 and anti-HIV-1-protease substances from Ganoderma lucidum. AB - A new highly oxygenated triterpene named ganoderic acid alpha has been isolated from a methanol extract of the fruiting bodies of Ganoderma lucidum together with twelve known compounds. The structures of the isolated compounds were determined by spectroscopic means including 2D-NMR. Ganoderiol F and ganodermanontriol were found to be active as anti-HIV-1 agents with an inhibitory concentration of 7.8 micrograms ml-1 for both, and ganoderic acid B, ganoderiol B, ganoderic acid C1, 3 beta-5 alpha-dihydroxy-6 beta-methoxyergosta-7,22-diene, ganoderic acid alpha, ganoderic acid H and ganoderiol A were moderately active inhibitors against HIV-1 PR with a 50% inhibitory concentration of 0.17-0.23 mM. PMID- 9862137 TI - Annojahnin from Annona jahnii: a possible precursor of mono-tetrahydrofuran acetogenins. AB - A new cytotoxic Annonaceous acetogenin, annojahnin (1), was isolated from the twigs of Annona jahnii (Annonaceae) by bioactivity-directed fractionation using lethality to brine shrimp. Compound 1 represents an unusual type of C-37 Annonaceous acetogenin, lacking either tetrahydrofuran (THF) or epoxide rings, bearing a keto group at C-10, and possessing a double bond located two methylenes away from a vicinal diol. The structure and absolute configuration of 1 were elucidated by 1H and 13C NMR, COSY, and single-relayed COSY and from chemical derivatives. 4-Deoxy-18/21-trans-annomontacin 10-one (4) and 4-deoxy-18/21-cis annomontacin-10-one (5), two semisynthetic mono-THF acetogenins, were prepared from 1 by reactions that mimic the biogenetic pathways. These acetogenins showed selective cytotoxicities, comparable or superior to adriamycin, among six human solid tumor cell lines. Reduction of the 10-keto of 1, to the racemic 10-OH derivative (3), retained the bioactivities as did the conversion of 1 to 4 and 5. PMID- 9862139 TI - Chelidocystatin, a novel phytocystatin from Chelidonium majus. AB - Greater celandine (Chelidonium majus L.) has traditional uses in European and Chinese herbal medicine. In the plant sap significant inhibitory activity against papain was observed. A cysteine proteinase inhibitor, named chelidocystatin, was isolated from the plant using papain Sepharose affinity chromatography followed by gel filtration and ion-exchange chromatography. Chelidocystatin showed a M(r) of 10,000 on SDS-PAGE with the pI of 9.3, and was a strong inhibitor of cathepsin L (Ki = 5.6 x 10(-11) M), papain (Ki = 1.1 x 10(-10) M) and cathepsin H (Ki = 7.5 x 10(-9) M). The complete amino acid sequence of the protein was obtained with N terminal sequencing and sequencing of the peptides after digestion of the protein. Moreover, a major part of the sequence was verified by molecular cloning. The conserved glycine residue at the N-terminal region and the QVVAG motif, which are both believed to be involved in the inhibitory activity, indicate that it is a member of the cystatin superfamily. The amino acid sequence of chelidocystatin shows a high degree of homology with cysteine proteinase inhibitors belonging to the phytocystatin group, especially with the recently described carrot and sunflower phytocystatins with which it shares 57% and 54% homology, respectively. PMID- 9862142 TI - Destruxin Ed1 a cyclopeptide from the fungus Metarhizium anisopliae. AB - A new destruxin Ed1 has been isolated from the entomopathogenic fungus Metarhizium anisopliae. Its structure was deduced from the NMR and mass spectral data. PMID- 9862131 TI - 3D HCCH-COSY-TOCSY experiment for the assignment of ribose and amino acid side chains in 13C labeled RNA and protein. AB - A new 3D HCCH-COSY-TOCSY experiment is presented for the assignment of RNA sugar and protein side chains. The experiment, which combines COSY and TOCSY units, is more powerful than the sum of individual HCCH-COSY and HCCH-TOCSY pulse sequences. The experiment was applied to a 13C, 15N-labeled 26 mer RNA complexed with the antibiotic tobramycin, and a 12 kDa 13C, 15N-labeled FKBP12 protein sample. The power of HCCH-COSY-TOCSY is demonstrated through complete spin system assignments of sugars in the 26 mer RNA sample, which could not be assigned using a combination of HCCH-COSY, HCCH-TOCSY and 13C-edited NOESY experiments. PMID- 9862144 TI - Immunohistochemical study of 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase in the rat cardiovascular system. AB - The enzyme complex 3beta-hydroxysteroid dehydrogenase (3beta-HSD) is involved in the biosynthesis of all classes of active steroids. It is known that the enzymatic activity of 3beta-HSD is present not only in classical steroidogenic tissues, but also in many peripheral tissues including cardiac tissue. To determine whether 3beta-HSD is present in rat non-steroidogenic tissues, we examined cardiovascular tissues including the ventricle, atrium, aortic arch, abdominal aorta, and inferior vena cava by immunohistochemistry and Western blotting using polyclonal antibody raised against a synthetic peptide of human placental 3beta-HSD. By Western blotting, protein bands immunoreactive for anti 3beta-HSD were detected at molecular weights of 42 and 37 kDa in both the ventricle and atrium, whereas only a 37 kDa band was recognized in both the aortic arch and abdominal aorta. By immunohistochemistry, immunoreactivity for 3beta,-HSD was detected in both the ventricular and atrial cardiocytes, while immunostaining was also found, though faintly, in the smooth muscles of the aortic arch, abdominal aorta, and inferior vena cava. These results suggest that cardiocytes may synthesize the steroidogenic 3beta,-HSD enzyme. PMID- 9862141 TI - Constituents and bioactive principles of Polygonum chinensis. AB - Isolation and characterization of the chemical constituents of Polygonum chinensis L. gave the new 25R-spirost-4-ene-3,12-dione. The known compounds stigmast-4-ene-3,6-dione, stigmastane-3,6-dione, hecogenin and aurantiamide acetate were also isolated from for the first time from this species. Their anti inflammatory and anti-allergic properties are described. PMID- 9862145 TI - The melanogenic system of Xenopus laevis. AB - Melanin pigments in lower vertebrates are often found in locations other than the skin, thus forming an extracutaneous pigmentary system of unknown function. The cellular and biochemical structure of this system is still poorly characterized. This paper deals with the ultrastructural and biochemical features of the melanogenic system of Xenopus laevis. Melanin containing cells were identified in the dorsal and ventral skin, and in the lung, spleen, liver and connective tissue surrounding blood vessels. The pigment cells in the skin and the lungs appeared to be typical melanocytes. The spleen contained isolated melanocyte-like cells, but most of the pigment cells present in this organ were associated with melanomacrophage centers. Conversely, the liver appeared devoid of melanocytes and only displayed melanomacrophage centers. Tyrosinase activity was found in all pigment-containing organs except the liver. All organs containing tyrosinase activity also displayed melanin formation potential from L-tyrosine. Therefore, tyrosine hydroxylase and melanin formation activities could be detected only in those organs containing typical melanocytes but not in locations such as the liver, where only melanomacrophages centers were found. PMID- 9862146 TI - Ouabain-like immunoreactivity in the medulla oblongata of rats. AB - An isomer of ouabain, the ouabain-like compound (OI,C), may participate in the regulation of body fluid volume and vascular tone. Forebrain regions, especially the hypothalamus, are reported to be sites of OLC action in the central nervous system. The medulla oblongata is another critical area involved in central cardiovascular regulation. We reported that the microinjection of either monoclonal antibody to ouabain T8B11 or Fab fragment of digoxin-specific antibody into the rostral ventrolateral medulla significantly decreased mean arterial pressure and renal sympathetic nerve activity in anesthetized normotensive rats (TERUYA et al.: J. Clin. Invest. 99: 2791-2798, 1997). Using T8B11, we examined the ouabain-like immunoreactivity in the medulla oblongata of normotensive rats. In periodate-lysine-paraformaldehyde fixed tissues, ouabain-like immunoreactive neurons were detected in the nuclei and regions in the medulla oblongata including the ventrolateral medulla, ventromedial medulla, nucleus ambiguus, caudal raphe nuclei, nucleus of solitary tract, and dorsal motor nucleus of the vagus. When an Fab fragment of digoxin-specific antibody was used as a first antibody, the digoxin-like immunoreactive neurons were distributed in almost the same pattern as those observed with the use of T8B11. In the brain fixed with the "three-step" procedure developed by YAMADA et al. (1987), which was used in a previous ouabain immunohistochemical study of the hypothalamus, ouabain-like immunoreactivity in the medulla oblongata was much weaker in intensity and less restricted in distribution than that in the hypothalamus. These findings suggest that ouabain-like immunoreactivities are present in the medulla oblongata with a manner of distribution different from that seen in the hypothalamus. Some ouabain immunopositive nuclei and regions in the medulla oblongata, especially the rostral ventrolateral medulla, may be other OLC action sites. PMID- 9862148 TI - Fine structure of hydrous chromosomes observed by low vacuum scanning electron microscopy. AB - Chinese hamster metaphase chromosomes were stained with platinum (Pt) blue and observed in the hydrous state with low vacuum scanning electron microscopes (LVSEM). The coiled structure of the chromatin fibers in the chromosomes was well recognized through the surrounding perichromosomal substances in the backscattered electron (BSE) mode at accelerating voltages of over 20 kV. Findings indicated that chromatin fibers in native chromosomes have a structure similar to the hierarchic coiled model. The present study also demonstrated that not only surface structures but also subsurface structures can be studied in the BSE mode of LVSEM, when the subsurface structures have been stained with heavy metal salts such as Pt blue. PMID- 9862147 TI - The innervation of taste buds in the soft palate and circumvallate papilla of the rat as revealed by the zinc iodide-osmium tetroxide technique. AB - The taste buds in the soft palate and the circumvallate papillae of the rat were investigated by the zinc iodide-osmium tetroxide technique. In addition, electron micrographs of taste buds stained with this method were presented for the first time. Differences in taste bud structures were found between the examined regions. The taste buds of the soft palate showed a complicated plexus of intragemmal nerve fibers. Some fibers exhibited terminal polymorphic swellings. Single branches could be traced close to the space of the taste pore. In the soft palate, the taste bud cells remained unstained, whereas in the circumvallate papillae of the tongue, a subpopulation of taste bud cells could be selectively stained and the intragemmal nerve fibers were characterized by large varicosities. The morphological dissimilarities between the taste buds of the investigated regions might be explained by their functional characteristics, or possibly their varying affinities to the taste qualities. Electron microscopic investigation of the stained circumvallate papillae revealed that the electron dense reaction product had primarily accumulated in a subpopulation of light cells. Dark cells exhibited only a slight labelling. In detail, the precipitate was found loosely distributed in the cytoplasm as well as the nuclei of the cells, and particularly concentrated at the membranes of light vacuoles, this probably being profiles of dilated endoplasmic reticulum. A few roundish accumulations of precipitate were seen in the cytoplasm of taste bud cells, which showed no intensive light microscopic staining. Labelled material was also found within the taste pores outside the apical processes of the cells. The present findings indicate that the zinc iodide-osmium tetroxide technique is applicable to neuroanatomical studies of taste buds. PMID- 9862149 TI - Immunohistochemical localizations of class II antigens and nerve fibers in human carious teeth: HLA-DR immunoreactivity in Schwann cells. AB - Nerve fibers and class II major histocompatibility complex (MHC) antigen expressing dendritic cells have been known to gather in the dental pulp beneath carious lesions. Significant functional interactions presumably occur between the neural and immune elements. The present study analyzed the morphological relationship between class II-expressing cells and nerve fibers in fuman carious teeth, visualized by a HLA-DR monoclonal antibody and a protein gene product 9.5 (PGP 9.5) polyclonal antibody; a confocal laser scanning microscope (CLSM) and an electron microscope were used. In pulps affected by early caries, HLA-DR-positive dendritic cells aggregated mainly in the cell-free zone associated with bundles of PGP 9.5-immuno-reactive nerve fibers. In pulps affected by advanced caries, the accumulated HLA-DR-positive cells and PGP 9.5-immunoreactive nerve fibers showed close association with each other especially beneath the odontoblast layer: the cells even embraced the nerve fibers. Intriguingly, class II molecules were recognized not only in dendritic cells but also in the Schwann cells of non myelinated nerves in the pulp. Using immuno-electron microscopy, class II molecules were localized on the surface of the non-myelinating Schwann cells and also within some vesicles, whereas myelinating Schwann cells lacked this immunoreactivity. PGP 9.5-immunoreactive nerve fibers were also observed densely in the odontoblast layer, and CLSM revealed an intimate association of the nerve fibers and dendritic cells. The immunoreactivity for HLA-DR in Schwann cells depended upon the severity of the carious lesion. Class II-expressing Schwann cells are suggested to function as antigen-presenting cells in addition to dendritic cells. PMID- 9862143 TI - Fine structural and morphometric studies on gastric parietal cells of peptic ulcer patients after long-term treatment with omeprazole. AB - Omeprazole, a substituted benzimidazole, is known to inhibit acid secretion from parietal cells in gastric glands, and is widely utilized as a drug for peptic ulcer. To clarify the ultrastructural changes in parietal cells from long-term treatment with a therapeutic dose of omeprazole, biopsy specimens of the gastric mucosa obtained from peptic ulcer patients were morphometrically analyzed before and after omeprazole treatment. Before treatment with omeprazole, parietal cells in both the stimulated and resting stages were observed; the stimulated cells possessed smaller amounts of tubulovesicles in the cytoplasm and numerous profiles of microvilli in the intracellular canaliculi, whereas the cells in the resting phase showed numerous profiles of tubulovesicles and poorly developed microvilli in the canaliculi. Eight weeks after the onset of omeprazole treatment, the amounts of both tubulovesicles in the cytoplasm and microvilli in the intracellular canaliculi drastically decreased. These decreases in the profiles of the membrane structures with a proton pump occurred concomitantly with a significant increase in autophagic vacuole/autolysosome-like structures. These results suggest that the membrane structures with proton pump are not recycled between tubulovesicles and microvilli of intracellular canaliculi in parietal cells after omeprazole treatment, but may be sequestrated into autophagosomes and degraded by lysosomal enzymes. PMID- 9862152 TI - Gamma aminobutyric acid immunoreactivity in the mouse adrenal gland during postnatal development. AB - To understand the developing processes of gamma-aminobutyric acid (GABA) containing chromaffin cells and nerve fibers in the mouse adrenal gland, we examined the tissues in various postnatal stages by immunohistochemistry using a GABA antibody. From birth until postnatal week 1, GABA-immunoreactivity was seen in very few nerve fibers, and in none of the chromaffin cells. At postnatal week 2, GABA-immunoreactivity appeared weakly in clusters of chromaffin cells and strongly in relatively numerous varicose nerve fibers. The immunoreactive nerve fibers were densely distributed in the small immunonegative chromaffin cells and large ganglion cells, but only sparsely so in the weak immunoreactive chromaffin cells. At postnatal week 3, the number of the immunoreactive chromaffin cells and nerve fibers further increased compared to that at postnatal week 2. The staining pattern of GABA-immunoreactive nerve fibers in the medullas was similar to that at postnatal week 2. From postnatal week 4 until postnatal week 8, the distribution and frequency of the immunoreactive chromaffin cells and nerve fibers were also similar to those at postnatal week 3. These results suggest that the expression of GABA in the chromaffin cells and in the nerve fibers of the mouse adrenal gland may be completed by postnatal week 3. PMID- 9862151 TI - Organization of nitric oxide-producing nerves in the rat pyloric sphincter. AB - The architecture of nitric oxide (NO)-producing nerves in the rat pylorus was studied in relation to the muscular structure. The musculature of the rat pylorus was observed to be composed of two discrete muscle loops (proximal and distal sphincters). Connective tissue septa containing neural elements divided the thick musculature of the distal sphincter into many bundles. The myenteric nerve plexus of the stomach with a subpopulation of NO-producing nerves was continuous with that of the duodenum. Nitrinergic nerve fibers which originated from the antral myenteric plexus ran through the connective tissue septa in the pyloric musculature and were densely distributed on the submucosal surface of the distal sphincter. The innermost portion of the distal sphincter consisted of smooth muscle cells showing many cytoplasmic processes and abundant nitrinergic nerve terminals. This particular architecture of the nitrinergic nerves in the sphincter would seem to account for the coordinate motor function of the rat pyloric sphincter. PMID- 9862150 TI - Distribution of F4/80-positive cells in developing ovaries in the mouse. AB - The mature ovary contains a large number of macrophages. In the present study, the distribution of macrophages in murine ovaries at various developmental stages was immunohistochemically studied using a monoclonal antibody against F4/80, a highly specific antigen of murine macrophages. The results showed that definite F4/80-positive stains were hardly detectable in ovaries on day 0 after birth. On day 7, a few F4/ 80-positive cells could be identified between the developing follicles. The positive stains were irregular in shape and showed little physical contact with the primordial or primary follicles. By days 14 and 21, when the theca cell layers of growing follicles were developing, the positive cells had extended or elongated to surround the cell layer. On day 28, besides the presence of elongating positive cells surrounding the growing follicles, irregularly shaped F4/80-positive cells became apparent in the interstitium between the growing follicles and also in the capsular tissues. Thereafter, positive cells with stellate appearance were detected in the corpora lutea, which first developed around 6 weeks of age. Although the positive cells were homogenously distributed in the corpora lutea in virgin adults, only a few sporadic positive cells were found there in pregnant mice. However, the positive cells infiltrated into the corpora lutea again in the postpartum period. These results show that ovarian macrophages exhibit dramatical changes in their distribution from neonatal to postpartum periods. PMID- 9862154 TI - Inferior vena cava filter placement for prevention of pulmonary tumor emboli of renal cancer with renal vein or vena caval tumor thrombus: prophylactic usage prior to transcatheter arterial embolization. AB - PURPOSE: To prevent pulmonary embolization of necrotic intravenous tumor thrombus after transcatheter embolization of renal cell carcinoma, we placed suprarenal inferior vena cava (IVC) filters. MATERIALS AND METHODS: Suprarenal IVC filters were placed prior to transcatheter arterial embolization (TAE) in four patients, two women and two men, with renal cancer accompanied by renal vein in one and vena caval tumor thrombus in three patients. We used one bird's nest filter for the patient with renal vein tumor thrombus and a titanium Greenfield filter for each of the three patients with vena caval tumor thrombus. TAE was performed with pure ethanol under balloon occlusion of the renal artery. RESULTS: IVC filters were successfully placed at the suprarenal position. The patients tolerated the procedure well and had extensive tumor infarction, including the tumor thrombus, on follow-up computed tomography (CT). No pulmonary infarction or adverse effects were seen during the follow-up period. One patient died from diabetic renal failure, two patients have been alive for 8.5 and 7.5 months respectively, the other one is lost to follow-up. CONCLUSIONS: Suprarenal IVC filter placement prior to TAE for advanced renal cell carcinoma invading the renal vein or the IVC is an effective procedure to prevent pulmonary thromboemboli and may contribute to longer survival. PMID- 9862156 TI - In-111 labeled leukocyte scintigraphy in patients with suspected inflammation after failed antibiotic therapy. AB - In-111 labeled leukocyte scintigraphy (In-111 WBC scan) was performed in 16 patients with inflammation suspected on the basis of laboratory findings, symptoms, and diagnostic imaging, but who had failed antibiotic therapy. In-111 WBC scans revealed an abnormal focus of radiotracer activity (positive scans) in five of 16 patients. No correlation was found between the peripheral WBC count and accumulation of In-111 WBC. Inflammatory disease suspected on the basis of the CRP level should be considered when In-111 WBC scanning results in negative findings. Our results indicated that In-111 WBC scanning has low sensitivity after antibiotic therapy. Selection of patients on the basis of persistent elevation of CRP may be valuable. PMID- 9862153 TI - MR appearance at different ages of osteoporotic compression fractures of the vertebrae. AB - PURPOSE: To elucidate the MR appearance at different ages of osteoporotic vertebral compression fractures. METHODS: We retrospectively analyzed sequential MR studies of 107 vertebrae in 75 patients with osteoporotic compression fractures. RESULTS: Vertebral deformities comprised 35 anteriorly wedged vertebrae, 65 fish vertebrae, four flat vertebrae, and others. Signal alterations were depicted as geographic areas of low intensity on T1-weighted images (T1-WI) and high intensity on T2*-WI, or linear areas of low intensity on T1-WI and high or low intensity on T2*-WI. In the acute stage, geographic alteration predominated. During a three-month period following the injuries, the extent of the geographic areas increased in 40% of cases on T1-WI. Geographic alteration declined in the chronic stage, whereas the prevalence of linear signals increased. Restoration of fatty marrow was preceded by linear signals without geographic signals on T1-WI (36%). The anteriorly wedged vertebrae collapsed faster than the fish vertebrae. CONCLUSIONS: The acute or subacute stage of compression fractures is characterized by a larger alteration of the geographic signal, whereas the chronic stage was evidenced by a smaller area of alteration of the geographic and linear signals or by restoration of fatty marrow. PMID- 9862158 TI - Thyroid dysfunction in children receiving neck irradiation for Hodgkin's disease. AB - Thyroid function was studied in 46 long-term survivors of pediatric Hodgkin's disease with a median follow-up time of 10.5 years. The mean age of the patients at the time of treatment was 8 years. Treatment consisted of radiotherapy alone in seven patients and combined radiation and chemotherapy in 39 patients. The radiotherapy dose to the thyroid gland was less than 2000 cGy in one, 2000-2500 cGy in 15, 2500-3000 cGy in 17, and greater than 3000 cGy in 13 patients. Evaluation consisted of clinical examination and thyroid function tests of total and free triiodothyronine, thyroxin, arid thyroid stimulating hormone levels. Twenty-one of 46 patients (45.6%) showed thyroid function abnormalities, however only nine of them had diffuse or nodular hyperplasia on physical examination. Risk factors of age, chemotherapy schema, total radiation dose, and dose per fraction did not significantly influence the incidence of thyroid dysfunction. PMID- 9862155 TI - Detection of paraumbilical or splenorenal shunts by intra-arterial-DSA in bleeding varices: another risk factor? AB - In six of 46 consecutive patients with liver cirrhosis and bleeding varices, selective intra-arterial digital subtraction angiography detected large spontaneous portosystemic shunts including three patent paraumbilical veins and three splenorenal shunts. Twenty-one patients died during a median 73-month follow up period after devascularization with (44 cases) or without (2 cases) esophageal transection. Univariate analysis of preoperative variables concerning hepatic reserve failed to show significant predictability on postoperative survival rate. Instead, the postoperative survival rate of patients with the decompressive shunts was significantly lower than that of patients without the shunt. In a subgroup of patients with well-compensated liver cirrhosis and bleeding varices, the coexistence of paraumbilical or splenorenal portosystemic shunt may be another potent variable predicting poor prognosis. PMID- 9862159 TI - A case of a solitary metastatic diaphragmatic tumor--relation to the peritoneal stomata of the diaphragm. AB - We herein describe a metastatic diaphragmatic tumor, an entity that has rarely been reported radiologically. The primary lesion was advanced colon cancer, and the root of the metastasis was thought to be the peritoneal stoma(ta) of the diaphragm, which has previously been shown to absorb both intraperitoneal fluid and small particles. PMID- 9862161 TI - A primary hepatic carcinoid tumor: evaluation by computed tomography and magnetic resonance imaging. AB - A case of primary hepatic carcinoid tumor is reported. A 40-year-old woman was admitted to our hospital because of an epigastric mass. Ultrasonography demonstrated a heterogeneous, hyperechoic mass with hypoechoic area in the left lobe of the liver. CT showed a hypodense lesion with more hypodense areas. Enhanced CT showed slight retention of contrast medium and a low density area that suggested necrosis in the tumor. The mass appeared as a heterogeneous area of low signal intensity with sharp margin on T1-weighted MRI. T2-weighted MRI revealed a high intensity mass with higher intensity foci. Left lateral segmental hepatic resection was performed. Immunohistochemical and electron microscopic studies revealed the carcinoid tumor. Postoperatively, careful, intensive search revealed no other primary source of the tumor. The higher intensity foci were microscopically identified as intratumoral hemorrhage, corresponding to hemorrhagic areas on the cut surface. Therefore, T2-weighted MRI might reflect the pathological characteristics of the tumor. PMID- 9862160 TI - A case of acute interstitial pneumonia indistinguishable from bronchiolitis obliterans organizing pneumonia/cryptogenic organizing pneumonia: high-resolution CT findings and pathologic correlation. AB - We report a case of histologically proved acute interstitial pneumonia (AIP) with subacute onset whose high-resolution CT (HRCT) findings were indistinguishable from those of bronchiolitis obliterans organizing pneumonia (BOOP)/cryptogenic organizing pneumonia (COP). The HRCT findings were air-space consolidation with air-bronchiologram associated with little ground-glass attenuation, and nodules. Some cases of AIP present HRCT findings indistinguishable from those of BOOP/COP. PMID- 9862157 TI - Three-dimensional power Doppler sonography of tumor vascularity. AB - PURPOSE: To assess the clinical value of three-dimensional (3D) power Doppler imaging of intratumoral blood flow. MATERIALS AND METHODS: Two hepatocellular carcinomas (HCC), one renal cell carcinoma, one hepatic focal nodular hyperplasia (FNH), and one iliac bone metastasis of HCC were used. Images were collected by power mode using the GE-Yokogawa LOGIQ 700. Power Doppler images were recorded on S-VHS videotape and fed into a workstation. Flow information was then extracted, and 3D images were constructed by surface rendering. RESULTS: Preparation of 3D images could be accomplished in one minute after feeding flow information into the workstation. Intratumoral vascular structure could be easily evaluated. This 3D imaging was useful for the differential diagnosis of hepatic tumors, especially HCC and FNH, which were confirmed in 3D imaging to have distinctly different intratumoral vascular structures. CONCLUSION: 3D imaging of intratumoral blood flow was possible by 3D processing of the power Doppler images. 3D power Doppler imaging is expected to be helpful for the differential diagnosis of tumors. PMID- 9862163 TI - Atypical acute reaction associated with radiotherapy: a case report. AB - Radiation therapy may cause acute and/or chronic skin reactions. In this paper a patient with contact urticaria associated with irradiation is described. We could not determine the agent behind the contact urticaria in our patient in light of the current literature. We are reporting this case because the literature neither mentioned radiotherapy as being among the agents that lead to contact urticaria nor reported contact urticaria as being among the acute reactions to radiotherapy. PMID- 9862162 TI - Abducens nerve enhancement in acute ophthalmoparesis. AB - Acute ophthalmoparesis (AO) is a monophasic disease characterized by acute onset of paresis of the extraocular muscles without ataxia or areflexia. Here we report a case of AO with gadolinium enhancement in the cisternal portion of the abducens nerves using contrast-enhanced three-dimensional magnetic resonance imaging. PMID- 9862164 TI - Fifteen cases of male breast carcinoma treated between 1980 and 1995. AB - Male breast carcinoma is a rare malignity. In Turkey, as in other countries, there are insufficient studies on male breast cancer. In the Radiation Oncology Department, Gulhane Military Medical Academy, we treated 15 cases of male breast cancer versus 1393 female breast cancer. Two of the cases had stage I, nine had stage II, and the remaining four had stage III disease. Median age was 52. All patients had definitive external beam radiation therapy with daily 200 cGy fractionation, following surgery. Median follow-up was 227 months. Five year survival rate was found to be 60% for all stages. Our evaluations of the pathologic findings, management, and treatment outcome were compared with literature. PMID- 9862165 TI - Percutaneous transluminal renal angioplasty of arterial divarication: application of kissing balloon technique. AB - We present a patient with renal artery stenoses due to fibromuscular dysplasia involving the point of arterial bifurcation that we could successfully dilate with percutaneous transluminal angioplasty. We used the kissing balloon technique to prevent possible complications of percutaneous transluminal renal angioplasty (PTRA), including occlusion of the other branch while dilating one branch. The kissing balloon technique worked well for PTRA of the arterial bifurcation. PMID- 9862167 TI - Introducing a downsized, open computer system with asynchronous transfer mode in PACS and its effect on image data transfer. AB - To assess the effectiveness of the introduction of a downsized, open computer system and asynchronous transfer mode (ATM) as a network in a medium-sized picture archiving and communication system (PACS), we conducted experiments on image data transmission. The speed of image transfer, including registration in the computers in the new PACS, was four to eleven times faster than that in our first PACS. However, the introduction of ATM did not contribute significantly to the improvement of speed of image transfer because of the use of Ethernet branches between the gateway and display workstation computers. A star architecture without Ethernet branches may be suitable for PACS using an ATM network. PMID- 9862168 TI - The VLDL receptor: an LDL receptor relative with eight ligand binding repeats, LR8. AB - The discovery in 1992 of a member of the low density lipoprotein receptor (LDLR) family with eight ligand binding repeats (LR8) has raised more questions than have been answered to date. Here, we summarize the current status of knowledge about this intriguing molecule, generally termed VLDL receptor, at the molecular biological, cell biological, and physiological levels. On one hand, the wealth of reports concerning the role(s) of this receptor in lipoprotein metabolism in mammalian systems has revealed partially conflicting details, particularly in regards to its natural ligand(s) and site of action. On the other hand, molecular genetic and biochemical studies in the chicken have clearly demonstrated the multiple roles of LR8 in the physiology and reproduction of egg-laying species, and have generated insights into the evolutionary aspects of the LDLR gene family. PMID- 9862169 TI - Metabolic modes of action of the statins in the hyperlipoproteinemias. AB - Conflicting results have been published during the past few years regarding the physiologic modes of action of the hydroxymethylglutaryl-CoA (HMG-CoA) reductase inhibitors, generally referred to as statins, using standard doses. Three mechanisms have been described: increased LDL catabolic rate, increased removal of LDL precursors resulting in decreased LDL production and decreased VLDL production. The physiologic effects of statins seem to depend on the underlying pathology of the disorders under therapy. More recent data using either the more potent atorvastatin or larger doses of previously available statins (e.g. simvastatin 80-160 mg/day), suggest that both the potency of the statins and the underlying pathopHysiology are important in determining the predominant physiologic responses of patients. To understand physiologic responses more completely, drug-dose-physiologic response curves of apo B kinetics in various groups of patients are needed. Simultaneous studies of apo B, triglycerides and cholesterol metabolism are also needed and are currently feasible. PMID- 9862166 TI - 99mTc-labeled chimeric anti-NCA 95 antigranulocyte monoclonal antibody for bone marrow imaging. AB - Chimeric mouse-human antigranulocyte monoclonal antibody (ch MAb) against non specific cross-reacting antigen (NCA-95) was labeled with 99mTc (using a direct method) and 125I (using the chloramine T method), and its binding to human granulocytes and LS-180 colorectal carcinoma cells expressing carcinoembryonic antigen on their surfaces, cross-reactive with anti-NCA-95 chimeric monoclonal antibody, increased in proportion to the number of cells added and reached more than 80% and 90%, respectively. In biodistribution studies, 99mTc and 125I labeled ch anti-NCA-95 MAb revealed high tumor uptake, and the tumor-to-blood ratio was 2.9 after 24 hours. The tumor-to-normal-organ ratio was also more than 3.0 in all organs except for the tumor-to-kidney ratio. Scintigrams of athymic nude mice confirmed the results of biodistribution studies that showed higher radioactivity in tumor and kidney of the mice administered with 99mTc-labeled ch MAb. A normal volunteer injected with 99mTc-labeled ch anti-NCA-95 antigranulocyte MAb showed clear bone marrow images, and a patient with aplastic anemia revealed irregular uptake in his lumbar spine, suggesting its utility for bone marrow scintigraphy and for the detection of hematological disorders, infections, and bone metastasis. PMID- 9862170 TI - In vitro production of beta-very low density lipoproteins and small, dense low density lipoproteins in mildly hypertriglyceridemic plasma: role of activities of lecithin:cholester acyltransferase, cholesterylester transfer proteins and lipoprotein lipase. AB - As a model for the formation of beta-very low density lipoproteins (VLDL) and small, dense LDL by the intraplasma metabolic activities in vivo, lipoproteins in fresh plasma were interacted in vitro with endogenous lecithin:cholesterol acyltransferase (LCAT) and cholesterylester transfer proteins (CETP) and subsequently with purified lipoprotein lipase (LpL). The LCAT and CETP reactions in a mildly hypertriglyceridemic (HTG) plasma at 37 degrees C for 18 h resulted in (1) esterification of about 45% plasma unesterified cholesterol (UC), (2) a marked increase in cholesterylester (CE) (+129%) and a decrease in triglyceride (TG) (-45%) in VLDL, and (3) a marked increase of TG (+ 341%) with a small net decrease of CE (-3.6%) in LDL, causing a significant alteration in the TG/CE of VLDL (from 8.0 to 1.9) and of LDL (from 0.20 to 0.93). The LDL in LCAT and CETP reacted plasma is larger and more buoyant than that in control plasma. In vitro lipolysis of control and LCAT and CETP-reacted plasma by LpL, which hydrolyzed >90% of VLDL-TG and about 50-60% of LDL-TG, converted most of VLDL in control plasma (>85%) but less than half (40%) of VLDL in LCAT and CETP-reacted plasma into the IDL-LDL density fraction and transformed the large, buoyant LDL in the LCAT and CETP-reacted plasma into particles smaller and denser than those in the control plasma. The remnants that accumulated in the VLDL density region of the postlipolysis LCAT and CETP-reacted plasma contained apo B-100 and E but little or no detectable apo Cs and consisted of particles having pre-beta and beta electrophoretic mobilities. The inhibition of LCAT during incubation of plasma, which lessened the extent of alteration in VLDL and LDL core lipids, increased the extent of lipolytic removal of VLDL from the VLDL density region but lowered the extent of alteration in the size and density of LDL. The LCAT, CETP and/or LpL-mediated alterations in the density of LDL in normolipidemic fasting plasma were less pronounced than that in mildly HTG plasma, but they became highly pronounced upon increase of its TG-rich lipoprotein level by the addition of preisolated VLDL or by the induction of postprandial lipemia. Although the effect of LCAT, CETP and LpL reactions in non-circulating plasma in vitro may be different from that in vivo, the above data suggests that the plasma TG-rich lipoprotein level and the extent of intraplasma LCAT, CETP, LpL and likely hepatic lipase (HL) reactions in vivo may play a role in determining the LDL phenotype. PMID- 9862171 TI - Branched synthetic peptide constructs mimic cellular binding and efflux of apolipoprotein AI in reconstituted high density lipoproteins. AB - This study investigates the suitability of the trimeric apolipoprotein (apo)AI(145-183) peptide that we recently described, to serve as a model to probe the relationship between apoAI structure and function. Three copies of the apoAI(145-183) unit, composed each of two amphipathic alpha-helical segments, were branched onto a covalent core matrix and the construct was recombined with phospholipids. A similar construct was made with the apoAI(102-140) peptide and used as a comparison with dimyristoylglycerophosphocholine (DMPC)-apoAI complexes. The DMPC-trimeric-apoAI(145-183) complexes had similar immunological reactivity with monoclonal antibodies directed against the 149-186 apoAI sequence (A44), suggesting that the A44 epitope is exposed similarly in both the synthetic peptide and the native apoAI complexes. The complexes generated with the trimeric apoAI(145-183) bind specifically to HeLa cells with comparable affinity to the DMPC apoAI complexes; they are a good competitor for binding of apoAI to both HeLa cells and Fu5AH rat hepatoma cells; finally, these complexes promote cholesterol efflux from Fu5AH cells with an efficiency comparable with the apo AI/lipid complexes. To study LCAT activation by the trimeric apo AI(145-183) construct, complexes were prepared with dipalmitoylphosphatidylcholine (DPPC), cholesterol (C) and either the trimeric construct or apoAI. LCAT activation by the trimeric construct was much lower than by apo AI, possibly because the conformation of the trimeric 145-183 peptide in DPPC/C/peptide complexes does not mimic that of apoAI in the corresponding complexes. In comparison, the complexes generated with the multimeric apoAI(102-140) construct had a poor capacity to mimic the physico-chemical and biological properties of apoAI. The apoAI(102-140) construct had low affinity for lipid compared with the (145-183) construct. After association with lipids, it was a poor competitor of DMPC-apoAI complexes for cellular binding and had only limited capacity to promote cholesterol efflux. These results suggest trimeric constructs can serve as an appropriate models for apoAI, enabling further investigations and new experimental approaches to determine the structure-function relationship of apoAI. PMID- 9862172 TI - Effect of probucol on serum lipids, atherosclerosis and toxicology in fat-fed LDL receptor deficient mice. AB - Although numerous transgenic mouse models for atherosclerosis have been developed recently, little is known about their response to hypolipidaemic or anti atherosclerotic agents. We investigated the effect of the known hypocholesterolaemic and anti-atherosclerotic drug probucol on serum lipids, lipoproteins and atherosclerosis in fat-fed low density lipoprotein (LDL) receptor deficient mice. Probucol at doses of 0.2 and 1% in the diet which are similar to those used in the mouse by other investigators reduced serum cholesterol by 26 and 37%, respectively. Probucol also reduced serum triglyceride levels by 33 and 47% at doses of 0.2 and 1%, respectively. The decrease in serum cholesterol and triglycerides was mainly due to a decrease of these lipids in VLDL and or chylomicrons. Despite these potentially beneficial changes in serum lipids atherosclerotic lesion areas in the aortic root were unchanged in the probucol treated mice. After 12 weeks treatment most of the mice receiving probucol had swollen feet and tails due to oedema. Histological examination of the base of the hearts from the probucol treated mice revealed lipid droplets within the reticuloendothelial and other interstitial cells. There was also an interstitial subacute inflammatory cell infiltration associated with the lipid deposition. The oedema induced by probucol could be the result of cardiac insufficiency due to interstitial lipidosis and inflammation in the base of the heart together with the extensive atherosclerotic lesions in the aortic sinus. PMID- 9862174 TI - The Gln-Arg 191 polymorphism of the human paraoxonase gene is not associated with the risk of coronary artery disease among Chinese in Taiwan. AB - Paraoxonase (PON1) is a high density lipoprotein-associated enzyme capable of hydrolyzing lipid peroxides, and thus, might protect lipoproteins from oxidation. A common polymorphism due to an amino acid substitution (Gln-Arg) at codon 191 is considered to be a major determinant of variation in serum PON1 activity. Recent studies have suggested that the PON1-191 polymorphism is an independent risk factor for coronary atherosclerosis in patients with or without diabetes mellitus. The association of PON1-191 polymorphism genotypes and coronary artery disease (CAD) among Chinese subjects in Taiwan was examined. The genotype of 218 angiographically documented CAD patients and the same number of age- and sex matched control subjects was determined. Genotypes AA, AB and BB were present in 25 (11%), 102 (47%) and 91 (42%) of control subjects, respectively, and in 30 (14%), 96 (44%) and 92 (42%) of CAD patients, respectively (chi2 = 0.57, P = 0.75 between groups). The frequency of the A allele was 0.36 for the control group and 0.35 for CAD patients (P = 0.94). No significant differences in the PON1-191 genotype frequencies could be found between groups when multivariate logistic regression analysis was performed, or different subgroups of age, sex or risk factors were analyzed. Among control subjects, there was also no significant difference between genotypes of the PON1-191 polymorphism and various clinical and lipid variables. In conclusion, our data suggest that there is no association between the Gln-Arg 191 polymorphism of the human PON1 gene and CAD among Chinese subjects in Taiwan. PMID- 9862173 TI - Low serum carotene level and increased risk of ischemic heart disease related to long-term arsenic exposure. AB - To elucidate the association between arsenic-related ischemic heart disease (ISHD) and serum antioxidant micronutrient level, residents aged 30 or older living in arseniasis-hyperendemic villages in Taiwan were recruited in a community-based health survey. A structured questionnaire was used to obtain a history of long-term exposure to arsenic through consuming artesian well water and fasting serum samples were also collected at the recruitment. A total of 74 patients affected with ISHD, who were diagnosed through both electrocardiography and Rose questionnaire interview, and 193 age-sex-matched healthy controls were selected for the examination of serum levels of micronutrients by high performance liquid chromatography (HPLC). There was a significant biological gradient between the risk of ISHD and the duration of consuming high-arsenic artesian well water. A significant reverse dose-response relationship with arsenic-related ISHD was observed for serum level of alpha- and beta-carotene, but not for serum levels of retinol, lycopene and alpha-tocopherol. Multivariate analysis showed a synergistic interaction on arsenic-related ISHD between duration of consuming artesian well water and low serum carotene level. An increased risk of arsenic-related ISHD was also associated with hypertension and elevated body mass index, but not with serum lipid profile, cigarette smoking and alcohol drinking. The findings seem to suggest that arsenic-related ISHD has a pathogenic mechanism which is at least partially different from that of ISHD unrelated to long-term exposure to arsenic. PMID- 9862175 TI - Ex vivo gene transfer of endothelial nitric oxide synthase to atherosclerotic rabbit aortic rings improves relaxations to acetylcholine. AB - Cholesterol feeding results in impaired endothelium dependent vasorelaxation. The role of nitric oxide in this process is unclear. The aim of this study was to evaluate the role of nitric oxide in cholesterol-induced vasomotor dysfunction by examining the effect of overexpression of eNOS in the hypercholesterolemic rabbit aorta on vascular reactivity. Vascular rings from the thoracic aorta of hypercholesterolemic rabbits were exposed ex vivo either to an adenoviral vector encoding endothelial nitric oxide synthase (AdeNOS) or Escherichia coli beta Galactosidase (AdbetaGal). Transgene expression was examined by histochemistry for beta galactosidase, immunohistochemistry for eNOS and cyclic GMP measurements and vasomotor studies were performed. Transgene expression was found to localize to the endothelium and adventitia. cGMP levels were significantly greater in AdeNOS compared to AdbetaGal transduced rings. Acetylcholine mediated relaxation was significantly impaired in cholesterol fed rabbits and was markedly improved by overexpression of eNOS. These results suggest that reduced NO bioavailability observed in cholesterol-induced vascular dysfunction can be partially overcome by eNOS gene transfer. PMID- 9862176 TI - Hepatic lipase mediates an increase in selective uptake of high-density lipoprotein-associated cholesteryl esters by human Hep 3B hepatoma cells in culture. AB - Selective uptake of high-density lipoprotein- (HDL-) associated cholesteryl esters (CE), i.e. lipid uptake independent from particle uptake, delivers CE to the liver and steroidogenic tissues in vivo. In vitro, besides hepatocytes and steroidogenic cells many other cell types selectively take up HDL CE. Hepatic lipase (HL) stimulates the internalisation of apoprotein (apo) B-containing lipoproteins by hepatocytes independent from lipolysis. In this study the role of HL in the hepatic metabolism of apo A-I-containing lipoproteins, i.e. HDL, was investigated. HDL3 (d = 1.125-1.21 g/ml) was radiolabeled in its protein (125I) and in its CE moiety ([3H]cholesteryl oleyl ether, ([3H]CEt)). HL originated from tissue culture media of hepatoma cells and from post-heparin plasma. Human Hep 3B hepatoma cells incubated in medium containing radiolabeled HDL3. In the absence of HL, the rate of apparent HDL3 particle uptake according to the lipid tracer ([3H]CEt) was in most cases in approximately 10-fold excess on that due to the protein label (125I), indicating selective CE uptake from HDL3. Addition of HL to these incubations increased the cellular uptake of [3H]CEt and of 125I from HDL3 and quantitatively the most prominent effect was an up to approximately 2.5-fold stimulation of apparent selective CE uptake ([3H]CEt-125I). This increase in selective CE uptake was observed in the presence of tetrahydrolipstatin, an inhibitor of the catalytically active site of HL, suggesting that this HL effect is independent from lipolysis. HL binds to cell surface heparan sulfate proteoglycans. To explore the role of these molecules for the HL effect on selective CE uptake, hepatoma cells were depleted of proteoglycans or Chinese hamster ovary (CHO) cells deficient in proteoglycan synthesis were used. Proteoglycan-deficiency reduced the HL-mediated increase in selective uptake by more than 80%. To investigate if low-density lipoprotein (LDL) receptors or the LDL receptor-related protein (LRP) are involved in the HL effect on selective CE uptake, murine embryonic fibroblasts (MEF) were used which are deficient in these receptors; alternatively, monensin, an inhibitor of endocytosis was present in the medium of Hep 3B cells during the uptake assay for labeled HDL3. These experiments yielded no evidence for a role of LDL receptors or LRP in the HL mediated increase in selective CE uptake. In summary, HL mediates an increase in HDL3 selective CE uptake by human Hep 3B hepatoma cells. This HL effect is independent from lipolysis and independent from LRP and LDL receptors. However this HL effect is susceptible to cell surface proteoglycan deficiency. The potential physiologic implication is that HL modifies HDL selective CE uptake by the liver in vivo and such an effect could play a role in reverse cholesterol transport. PMID- 9862177 TI - Effect of human apolipoprotein E isoforms on plasma lipids, lipoproteins and apolipoproteins in apolipoprotein E-deficient mice. AB - The current study compared the acute effect of human apolipoprotein (apo) E isoforms on plasma lipids, lipoproteins and apolipoproteins 6 h after a bolus intravenous injection of individual isoforms into apo E-deficient mice. We found a large accumulation of remnant particles not only in the d<1.019 g/ml fraction but also in the d = 1.019-1.063 mg/dl fraction in the setting of absence of endogenous mouse apo E. A significant reduction in total cholesterol (49, 47 and 18%) (P<0.005), cholesterol in the d<1.019 g/ml fraction (56, 50 and 18%) and in the d = 1.019-1.063 mg/dl fraction (38, 40 and 17%) was obtained with apo E-3, E 4 and E-2, respectively. Apo E-3 and E-4 showed more pronounced total cholesterol lowering effect than E-2 (P<0.0001). In the d<1.019 g/ml fraction, apo E-3 and E 4 resulted in a marked decrease in apo B-100 (36 and 34%), B-48 (48 and 52%), A-I (48 and 44%) and A-IV (52 and 46%), respectively. The decrease caused by apo E-2 in apo B-100 (19%), B-48 (16%), A-I (18%) and A-IV (33%) was less than that of E 3 or E-4. In the d = 1.019-1.063 g/ml fraction, an apparent decline in apo B-48 (42 and 38%), A-I (39 and 40%) and an increase in apo B-100 (25 and 18%) were observed after apo E-3 and E-4 injection, respectively, while apo E-2 did not cause an appreciable change in these apolipoproteins (-4 to 6%). Compared to normal saline, liver total cholesterol content was increased by 37, 34 and 16% (P<0.05) after apo E-3, E-4 and E-2 injection, respectively. Apo E-3 and E-4 showed the same high affinity binding to mice hepatic LDL receptor, while apo E-2 was severely defective in binding. These findings indicate that apo E polymorphism is an important factor modulating remnant lipoprotein metabolism. PMID- 9862178 TI - Nitric oxide, prostacyclin and cyclic nucleotide formation in externally stented porcine vein grafts. AB - Non-restrictive, porous, external stents inhibit neointima formation in porcine vein grafts. Since the mechanisms underlying these effects are unknown we investigated the impact of this external stent on factors known to inhibit vascular smooth muscle cell proliferation: prostacyclin (PGI2), nitric oxide (NO), cAMP and cGMP formation in different regions of stented and unstented porcine vein grafts. Paired stented and unstented saphenous vein-carotid artery interposition grafting was carried out in Landrace pigs. One month after surgery, the vessels were excised and the formation of PGI2, cAMP and cGMP determined using radioimmunoassay and nitric oxide synthase (NOS) distribution studied using autoradiography and histochemistry. There were no significant differences between PGI2, cAMP and cGMP (nitroprusside-stimulated) formation in the medial/intimal regions of grafts of stented vein graft and ungrafted saphenous vein whereas all were significantly reduced in unstented vein graft. A23187-stimulated cGMP formation (mediated by NO release) and NOS content was significantly greater in the medial/intimal region of stented and unstented vein graft compared to ungrafted saphenous vein, indicating induction of endothelial NOS (eNOS) in both types of graft. This normalisation of the PGI2-cAMP axis and guanylyl cyclase activity in the medial/intimal region may contribute to the beneficial impact of the external stent on vein graft thickening. The increase in eNOS in both stented and unstented vein grafts mitigates against this isoform as playing a role in mediating the inhibitory effect of the stent on neointima formation. In the adventitia of both stented and unstented grafts there was an increase in PGI2, cAMP and cGMP formation compared to ungrafted saphenous vein, the production being greater in the stented compared to the unstented graft. In the adventitia of stented veini grafts, NOS, detected with NAPDH diaphorase staining, was associated with microvessels as well as with inflammatory cells. Taken together, these data are suggestive of a role for PGI2 and NO in promoting microangiogenesis in the adventitia of stented vein grafts which may in turn minimize graft hypoxia, an established contributory factor to neointima formation. PMID- 9862179 TI - Magnitude of alimentary lipemia is related to intima-media thickness of the common carotid artery in middle-aged men. AB - Fat intake leads to generation of potentially atherogenic triglyceride-rich lipoproteins (TRL). To investigate the relationship between early atherosclerotic changes and accumulation of hepatic and intestinal TRL after oral fat intake, an estimate of the intima-media thickness (IMT) was made using ultrasound of the common carotid artery, and postprandial TRL was quantified during a standardized oral fat tolerance test in 30 healthy normo- and hypertriglyceridemic middle-aged men. At base line the expected positive association between the LDL cholesterol level and the IMT of the common carotid artery was observed (r = 0.53, P<0.01). In addition, postprandial plasma triglycerides, in particular those measured late (6 h) after intake of the test meal, correlated positively with the IMT (r = 0.44, P<0.05). Of note, this latter correlation was independent of both the LDL cholesterol and the fasting plasma triglyceride concentrations. In a multivariate analysis, 39% of the total variability for the common carotid IMT were explained by age, LDL cholesterol and the postprandial triglyceride level. In univariate analysis, few statistically significant relations were found between common carotid IMT and postprandial levels of chylomicron remnants, VLDL and VLDL remnants of different particle size, the latter determined by specific measurements of ApoB-48 and ApoB-100 in subfractions of TRL. Therefore, in healthy middle-aged men, elevated postprandial triglyceride levels might identify a metabolic state related to early atherosclerosis. PMID- 9862181 TI - Effect of the fat composition of a single meal on the composition and cytotoxic potencies of lipolytically-releasable free fatty acids in postprandial plasma. AB - Ingestion of a meal increases plasma levels of triglyceride (TG)-rich lipoproteins through the secretion of intestine-derived chylomcirons and liver derived very low density lipoproteins (VLDL). We have determined the effects of the fat composition of a single meal on the composition of TG in TG-rich lipoproteins (VLDL + chylomicrons) and circulating and lipolytically-releasable free fatty acids (FFA) in postprandial (PP) plasma and on the cytotoxic potencies of the lipolytically-released FFA to cultured arterial wall cells. PP lipemia was induced by feeding fasted normolipidemic human subjects with a meal rich in saturated fat (SF) and another meal rich in polyunsaturated fat (PUF), or vice versa; each meal provided 65% of energy as fat, and polyunsaturated to saturated fatty acid ratios (P/S) of the SF and PUF in the meals were 0.40 and 2.49, respectively. The mean P/S of TG in TG-rich lipoproteins (1.43) and circulating FFA (1.46) in 4 h PP plasma of PUF were significantly higher than those in PP plasma of SF (0.44 and 0.59, respectively) in fasting plasma (0.52 and 0.53, respectively). In vitro lipolysis of fasting and PP serum by purified bovine milk lipoprotein lipase (LpL) resulted in a marked (8.8-12.3-fold) increase in the serum FFA level. The P/S of serum FFA in postlipolysis fasting and PP serum were consistently higher than that of FFA or that of TG associated with TG-rich lipoproteins in prelipolysis fasting and PP serum, indicating that polyunsaturated TG in VLDL and/or chylomicrons is more susceptible than saturated TG to lipolysis. When postlipolysis serum was interacted with cultured endothelial cells and mouse peritoneal macrophages (MPM), the lipolytically released FFA in PP serum of SF and PUF disrupted the barrier function of endothelial cells and were cytotoxic to cultured MPM; FFA in postlipolysis fasting serum was not cytotoxic. FFA in postlipolysis PP serum of PUF were consistently more potent than that in postlipolysis PP serum of SF. Further study showed that all long-chain monounsaturated FFA and polyunsaturated FFA, but not saturated FFA, incorporated into lipoproteins (LDL) were cytotoxic to cultured MPM. In conclusion, despite the generally well-accepted belief that SF is more atherogenic than PUF, the present study provides in vitro evidence that the lipolytic remnant products of TG-rich lipoproteins produced after a meal rich in PUF are more injurious to arterial wall cells than those produced after a meal rich in SF. PMID- 9862180 TI - A common methylenetetrahydrofolate reductase gene mutation and longevity. AB - Homozygotes (TT genotype) for the C677T mutation in the gene of methylenetetrahydrofolate reductase (C677T/MTHFR mutation) constitute about 12% of the Caucasian population. They have mild hyperhomocysteinemia which is an established risk factor for cardiovascular disease. If the mutation is associated with premature death its prevalence is expected to be lower in the elderly than in the young. To test this we determined the C677T/MTHFR genotypes in 220 newborn and 222 elderly 80-108-year-old Swedes. In the newborn and elderly, the allele frequency, of the C677T/MTHFR mutation was 29.1 and 27.0% and the mutant homozygote frequency was 10.0 and 9.5%, respectively. In a meta analysis of the present and three previous studies including a total of 1388 elderly and 1415 younger subjects, the odds ratio (OR) representing the likelihood of the TT genotype to attain old age relative to the CC genotype was 0.87 (95% confidence interval (CI), 0.69-1.11) and relative to both the CC and CT genotypes was 0.83 (95% CI, 0.66-1.04). This finding does not suggest that the C677T/MTHFR mutation is a strong risk factor for diseases frequently leading to premature death. PMID- 9862182 TI - Lipoprotein(a) interactions with lipid and non-lipid risk factors in patients with early onset coronary artery disease: results from the NHLBI Family Heart Study. AB - BACKGROUND: A positive interaction between high plasma lipoprotein(a) [Lp(a)] and unfavorable plasma lipid levels has been reported to result in very high risk for premature coronary artery disease (CAD). We further examined this issue for men and women with early onset CAD. We also examined potential interactions between Lp(a) and non-lipid risk factors. METHODS AND RESULTS: In 338 men and women with early onset CAD (most with a positive family history of early CAD) and 480 general population controls, we measured Lp(a), lipids and other risk factors. In univariate analysis, relative odds for CAD was 1.7 (P = 0.002) for plasma Lp(a) >50 mg/dl. Elevated Lp(a) level was found to interact with adjusted plasma total/high density lipoprotein (HDL) cholesterol such that when Lp(a) was over 50 mg/dl and adjusted plasma total/HDL cholesterol >5.8, relative odds for CAD were 8.0-9.6 (P<0.0001) in multiple logistic regression. Non-lipid risk factors were generally found to multiply the risk associated with Lp(a) (as predicted by logistic regression) without evidence for interaction. CONCLUSIONS: We find evidence that Lp(a) does interact positively with adjusted plasma total/HDL cholesterol ratio. Aggressive risk factor intervention, especially for lipids, in those with elevated Lp(a) therefore appears indicated. PMID- 9862183 TI - Cholesterol oxides and natural autoantibodies. PMID- 9862184 TI - p53 mutations, benzo[a]pyrene and lung cancer: a reply. PMID- 9862185 TI - Comet assay responses as indicators of carcinogen exposure. AB - Over 200 agents/factors have been examined in the single cell gel electrophoresis assay, more commonly known as the Comet assay, performed either in vitro or in vivo in a variety of species. Unequivocal carcinogenicity data are available for 119 of them, amongst which unequivocal Comet assay data exist for 95 agents. Of these 95 agents the prevalence of carcinogens was 88% (84/95). The carcinogens that were Comet positive (sensitivity) formed 88% (74/84), the non-carcinogens that were Comet negative (specificity) formed 64% (7/11). This simple analysis of the Comet assay has not taken account of the difference between in vitro and in vivo responses, species differences or organ and tissue differences. Also, limitations as to the conduct of the assay have not been examined in any depth. Thus, at the present time the Comet assay has high sensitivity for carcinogens, but its specificity is uncertain because few non-carcinogens have been tested. PMID- 9862186 TI - Genetic toxicology of abused drugs: a brief review. AB - Although numerous studies have been conducted on abused drugs, most focus on the problems of addiction (dependence) and their neurotoxicities. Now accumulated data have demonstrated that the genotoxicity and/or carcinogenicity of abused drugs can also be detrimental to our health. In this review, commonly abused substances, including LSD, opiates (diacetylmorphine, morphine, opium and codeine), cocaine, cannabis, betel quid and khat, are discussed for their potential genotoxicity/carcinogenicity. The available literature in the field, although not as abundant as for neurotoxicity, clearly indicates the capability of abused drugs to induce genotoxicity. PMID- 9862187 TI - Diazepam induces meiotic delay, aneuploidy and predivision of homologues and chromatids in mammalian oocytes. AB - The tranquilizer and anti-convulsant diazepam (DZ) is a suspected aneugen. In order to assess its aneugenic potential in mammalian oogenesis we exposed in vitro maturing mouse oocytes to the drug. Spindle formation and cell cycle progression, the behaviour of chromosomes and the distribution of mitochondria were characterized with respect to induction of numerical chromosomal aberrations. A concentration of 25 microg/ml DZ induced a pronounced delay in maturation and blocked a high percentage of oocytes in meiosis I. This arrest was partly reversible. Hyperploidy was slightly increased in oocytes matured in the presence of 5 microg/ml DZ and became significantly elevated in oocytes matured with 25 microg/ml DZ, relative to controls. Concomitantly, DZ induced spindle aberrations and displacement of chromosomes from the equator, but unlike in mitosis and in male meiosis most oocytes still possessed bipolar spindles. A significant fraction of meiotically delayed, metaphase I-blocked oocytes exposed to 25 microg/ml DZ contained univalents. Some DZ-treated oocytes progressing to meiosis II exhibited one or multiple single chromatids. Precocious chiasma resolution and equational segregation of chromatids from functional univalents in first anaphase (predivision) may be responsible for this condition, a mechanism also discussed in the aetiology of maternal age-related aneuploidy. DZ disturbed the spatio-temporal distribution of mitochondria during oocyte maturation, possibly by binding to peripheral-type benzodiazepine receptors on mitochondria, thus affecting the availability of ATP and calcium homeostasis. Blocks in maturation may also relate to binding of DZ to calmodulin. Data suggest that DZ exposes mammalian oocytes to predivision and aneuploidy. Thresholds, long lasting effects of DZ in vivo and sex-specific sensitivities in chemically induced aneuploidy of mammalian germ cells are critically evaluated. PMID- 9862188 TI - Detection of a mouse H-ras codon 61 mutation using a modified allele-specific competitive blocker PCR genotypic selection method. AB - A modified allele-specific competitive blocker PCR (ACB-PCR) has been developed as an approach for genotypic selection, the detection of a rare mutant allele based solely upon its altered nucleotide sequence. ACB-PCR genotypic selection operates through the preferential PCR amplification of mutant DNA using a primer that has more mismatches to the wild-type allele than the mutant allele. In addition, a blocker-primer with a 3'-terminal dideoxynucleotide and more mismatches to the mutant allele than the wild-type allele is incorporated to reduce the background and increase sensitivity. Using ACB-PCR, the CAA-->AAA base substitution at codon 61 of the mouse H-ras gene was detected regularly at mutant fractions of 10(-5). To accurately quantify the occurrence of this particular mutation, an internal amplification standard (AS) DNA was constructed. The H-ras and AS DNAs were subject to the same genotypic selection but were amplified using different upstream primers to give PCR products that can be distinguished by size. Defined mixtures of mutant and wild-type AS DNAs were used to study the effects of various components of the ACB-PCR. The concentration of dNTPs, blocker primer and Perfect Match Polymerase Enhancer, as well as the choice of thermostable DNA polymerase and annealing temperature were examined. Conditions were identified for the concurrent detection of the CAA-->AAA mutation in the H ras and AS DNAs. Using the identified conditions, approximately equal signals were obtained from equivalent amounts of the two DNA templates over a wide range of mutant fractions (1 in 10 to 1 in 10(5)). This ACB-PCR method can be used for any application where it is necessary to quantify relatively small mutant fractions. PMID- 9862189 TI - Formation of 8-oxoguanine in cellular DNA of Escherichia coli strains defective in different antioxidant defences. AB - This paper examines the relationship in Escherichia coli between the in vivo content of 8-oxoguanine (8-oxoG) in chromosomal DNA and deficiencies of various key antioxidant defences. The structural genes for catalases (katG and katE), cytosolic superoxide dismutases (sodA and sodB) or formamidopyrimidine-DNA glycosylase (fpg) were inactivated to obtain bacterial strains lacking the scavenger enzymes for H2O2 or O2.- or the DNA repair protein for 8-oxoG. Wild type bacteria showed 5-fold increased sensitivity to both lethality and mutagenesis by H2O2 in K medium (1% casamino acids and 1% glucose), as compared with nutrient broth. This higher sensitivity was associated with increased chromosomal oxidative damage, estimated as the 8-oxodG content, and with a marked decrease in both catalase and SOD activities. Bacteria lacking both cytosolic SODs (sodA sodB mutant) displayed increased 8-oxodG content in chromosomal DNA (2.8-fold that of the wild-type) when grown under standard aerated conditions. Comparatively, no significant difference in 8-oxodG content was observed in cells grown without aeration. Bacteria totally devoid of catalase activity (katG katE mutant) showed wild-type contents of 8-oxodG in chromosomal DNA when grown under aerated conditions. Nevertheless, the protective role of catalase in preventing formation of 8-oxodG in chromosomal DNA became evident under oxidative stress conditions: growth under hyperoxygenation and, particularly, following H2O2 exposure. Catalase deficiency resulted in a dramatic decrease in viability after H2O2 exposure. A deficiency of Fpg protein also sensitized E.coli to H2O2 lethality, though to lesser extent than a deficiency of catalase activity. However, the scavenger enzyme and the DNA repair protein protected equally against 8-oxoG formed in vivo upon H2O2 treatment. PMID- 9862190 TI - Sensitivity of group F xeroderma pigmentosum cells to UV and mitomycin C relative to levels of XPF and ERCC1 overexpression. AB - The XPF and ERCC1 proteins form a tight complex and function as an endonuclease to incise on the 5'-side of pyrimidine dimers in DNA. Levels of both proteins are extremely low in group F xeroderma pigmentosum (XP-F) cells. We transfected XP-F cells with the plasmids expressing XPF or ERCC1 and examined levels of both proteins in the cells. Although XP-F cells are sensitive to UV and mitomycin C (MMC), cells overexpressing XPF expressed ERCC1 as well and resistance to UV and MMC was restored to the normal level. In contrast, cells overexpressing ERCC1 did not express XPF and were still sensitive to UV and MMC. These results indicate that both the XPF and ERCC1 proteins are required to repair UV- and MMC-induced DNA damage. Even though a high level of ERCC1, which has been presumed to be a catalytic subunit of the endonuclease, is stably present in XP-F cells, ERCC1 protein alone cannot carry out excision repair completely. PMID- 9862191 TI - Genetic analysis of PHIP intestinal mutations in MutaMouse. AB - The mutagenicity of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) was investigated in male MutaMouse mice administered 20 mg/kg per o.s. for 4 days and killed 7 days later. Genomic DNA was extracted from liver, kidney and small and large intestine and the mutation frequency (MF) at the lacZ locus was determined using a positive selection assay. Mutant lacZ clones from the intestine were characterized further by direct PCR amplification and DNA sequencing. A total of 57 lacZ mutants from PhIP-treated (40) and untreated (18) mice were analysed. In mutants from the PhIP group, 33% were G:C-->T:A transversions from a total of 65% base substitutions (cf. 17% in the vehicle control group). In untreated control mice, 39% of mutants were G:C-->A:T transitions from a total of 72 % base substitutions (cf. 25 % in the PhIP group). Interestingly, 20% of the PhIP group mutations were due to G:C base pair (-G) deletions (cf. none in controls). This study confirms that PhIP is mutagenic to the intestine of the MutaMouse and induces a spectrum of mutations which are clearly distinct from those spontaneously generated. Also, the PhIP mutation signature in vivo is very similar to that observed for the HPRT and DHFR loci in hamster and human cells in vitro. This suggests that the mutational characteristics of PhIP are well conserved over different reporter genes and between species and that the mutation signature could be of value in molecular epidemiology studies. PMID- 9862192 TI - Mutation studies in lacI transgenic mice after exposure to radiation or cyclophosphamide. AB - We have used the Big Blue lacI transgenic mouse reporter system to investigate mutation induction in the testes, spleen and liver after exposure to an internally incorporated radionuclide, 114mIn, whole body irradiation with 60Co gamma-rays and systemically administered cyclophosphamide. Spontaneous mutation frequencies were 6-17x10(-6). No statistically significant mutation induction was observed in testes or spleen at 35 days after exposure to any test agent, although mutation frequencies tended to be increased (by approximately 1.5-fold) after exposure to 1 Gy gamma-rays. However, liver mutation frequencies were doubled after treatment with 100 mg/kg cyclophosphamide and were elevated by approximately 2.5-fold after systemic administration of 114mIn and 4.5-fold after 1 Gy 60Co gamma-rays. When data from all organs were pooled, mutation frequency was doubled after exposure to 1 Gy gamma-rays, but no other significant increases were observed. These findings support the hypothesis that the lacI transgenic mouse may be relatively inefficient at detecting mutations induced by exposure to ionizing radiation or other agents which produce a spectrum of deletion sizes, including those which are larger than the lacI transgene. PMID- 9862193 TI - Spontaneous mutation during fetal development and post-natal growth. AB - Somatic mutations seem to accumulate slowly with age during adult life in both mice and men. There is, however, a substantial mutant frequency at birth, suggesting that the rate of accumulation is much higher before birth. This suggests that DNA replication plays an important role in the generation of spontaneous mutations. Since most cell division and accompanying DNA replication occurs early in development, more mutations would arise during growth and development. Indeed, if the mutations are genetically neutral, the mutant frequency would rise very rapidly during early fetal growth, more slowly during later fetal growth and development and still more slowly after birth. To test this hypothesis, we have assayed the mutant frequencies from before birth to 28 days after birth, by which time most growth has occurred. We have used the F1 mice generated by crossing SWR females and MutaMouse males. The MutaMouse has a rescuable lacZ/lambda shuttle vector that can be assayed for an in vivo mutation in an in vitro system. Up to and including birth we assayed the entire animal for mutants; at 14 and 28 days after birth we assayed the small intestine. The data show that, as expected, many mutations arise early in development, by 12.5 days after conception, and confirms the non-linearity of mutation with age. In these mice, about one third of mutations arise before birth, about one third during growth to adulthood and the remaining during the rest of the animal's life, although this depends somewhat on the tissue. PMID- 9862194 TI - Comparative mutagenic and genotoxic effects of three antimalarial drugs, chloroquine, primaquine and amodiaquine. AB - Comparative mutagenic and genotoxic effects of three antimalarial drugs, chloroquine, primaquine and amodiaquine, were assessed in the Ames mutagenicity assay (in strains TA97a, TA100, TA102 and TA104) and in vivo sister chromatid exchange (SCE) and chromosome aberration (CA) assays in bone marrow cells of mice. These are the most commonly used antimalarial drugs available at present throughout the world. The results of the bacterial mutagenicity assays showed a very weak mutagenic effect of all three drugs in Salmonella strains TA97a and TA100 both with and without S9 mix and in TA104 only with S9 mix. The results of the in vivo SCE and CA assays indicate that these three drugs are genotoxic in bone marrow cells of mice. PMID- 9862195 TI - Specific mutational spectrum of dimethylnitrosamine in the lacI transgene of Big Blue C57BL/6 mice. AB - Dimethylnitrosamine (DMN) produces tumors in mice predominantly in the liver, but also in the kidney and lung. It forms O6-methylguanine adducts in DNA, which induce G:C-->A:T transitions. We have analyzed the spectra of spontaneous and DMN induced mutations in the lacI transgene of the Big Blue mouse (C57BL/6). In both cases, mutations in the liver, kidney and lung were predominantly base substitutions, among which G:C-->A:T transitions were the most frequent. In contrast, a high incidence of short deletions (2-23 bp) was only found in the liver of treated mice. The deletions often occurred at direct repeat sequences. Single-base deletion incidence was also higher in the liver than in the kidney and lung. These results imply that accumulation of DNA lesions or their repair in liver is different from other organs. Spontaneous and induced base substitutions and deletions appeared to be randomly distributed in the lacI gene and an apparent hotspot was not observed, except for a 4 bp deletion of a (TGGC)3 sequence at positions 621-632. The present data demonstrate, for the first time, that DMN induces short deletions especially in the liver, although the mechanism involved needs further investigation. PMID- 9862196 TI - Contribution of theafulvins to the antimutagenicity of black tea: their mechanism of action. AB - Theafulvins were isolated from black tea aqueous infusions and their antimutagenic activity was evaluated against a number of food carcinogens. Theafulvins gave rise to a concentration-dependent inhibition of the mutagenicity of 2-amino-3-methylimidazo-[4,5-f]quinoline, 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine, benzo[a]pyrene, 7,12-dimethylbenz[a]anthracene, nitrosopyrrolidine and nitrosopiperidine, but, in contrast, the mutagenicity of aflatoxin B1 was enhanced. The mutagenicity exhibited by N'-methyl-N'-nitro-N-nitrosoguanidine and 9-aminoacridine was not influenced and weakly inhibited by theafulvins, respectively. The p-hydroxylation of aniline and the O-dealkylations of methoxy-, ethoxy- and, to a lesser extent, pentoxyresorufin were inhibited by theafulvins in a concentration-dependent manner. When microsomal metabolism was terminated after metabolic activation of the promutagens, incorporation of the theafulvins into the activation system did not modulate the mutagenic response. It is concluded that theafulvins play an important role in the antimutagenic activity of black tea by inhibiting cytochrome P450-dependent bioactivation of the carcinogens. PMID- 9862197 TI - SOS induction by gamma-radiation in Escherichia coli strains defective in repair and/or recombination mechanisms. AB - Ionizing radiation causes several types of DNA lesions, mainly single- or double strand breaks and base damage. By means of the chromotest, an assay that allows the level of the SOS response to be monitored via beta-galactosidase enzymatic activity, the roles of several repair (uvrA, recN and oxyR) and recombination (recB, recJ and recO) genes in the response of Escherichia coli to gamma radiation were studied. The results indicate that all the repair- and recombination-deficient strains were more sensitive to the lethal effects of ionizing radiation. However, the SOS activation pattern was somewhat different. The minimal inducing dose in uvrA and recN mutants was lower than in the wild type, whereas their SOS response was higher at all doses. Conversely, in the strains lacking an active recB, recJ or recO gene, the doubling dose was almost the same as in the wild-type but the level of induction remained stable over a wide dose range. These findings suggest that neither single- nor double-strand breaks are in themselves direct SOS inducers and that while uvrA, recN and oxyR take part in different repair or protective pathways, apparently recB, recJ and recO participate in damage processing leading to SOS induction, as well as in recombination repair. PMID- 9862198 TI - Urinary and serum mutagenicity studies with rats implanted with depleted uranium or tantalum pellets. AB - During the 1991 Persian Gulf War several US military personnel were wounded by shrapnel fragments consisting of depleted uranium. These fragments were treated as conventional shrapnel and were not surgically removed to spare excessive tissue damage. Uranium bioassays conducted over a year after the initial uranium injury indicated a significant increase in urine uranium levels above natural background levels. The potential mutagenic effects of depleted uranium are unknown. To assess the potential mutagenic effects of long-term exposure to internalized depleted uranium, Sprague-Dawley rats were implanted with depleted uranium and their urine and serum were evaluated for mutagenic potential at various times after pellet implantation using the Ames Salmonella reversion assay. Tantalum, an inert metal widely used in prosthetic devices was used for comparison. Enhancement of mutagenic activity in Salmonella typhimurium strain TA98 and the Ames II mixed strains (TA7001-7006) was observed in urine samples from animals implanted with depleted uranium pellets. In contrast, urine samples from animals implanted with tantalum did not show a significant enhancement of mutagenic activity in these strains. In depleted uranium-implanted animals, urine mutagenicity increased in a dose- and time-dependent manner demonstrating a strong positive correlation with urine uranium levels (r = 0.995, P < 0.001). There was no mutagenic enhancement of any bacterial strain detected in the sera of animals implanted with either depleted uranium or tantalum pellets. The results suggest that uranium content in the urine is correlated with urine mutagenicity and that urinary mutagenicity might be used as a biomarker to detect exposure to internalized uranium. PMID- 9862199 TI - Cytogenetic characterization of the transgenic Big Blue Rat2 and Big Blue mouse embryonic fibroblast cell lines. AB - The transgenic Big Blue Rat2 and Big Blue mouse embryonic fibroblast cell lines have been used to complement the transgenic Big Blue rat and mouse in vivo mutagenesis assays. However, limited information is available regarding the karyology of these cell lines. Therefore, we have characterized the ploidy, mitotic index, spontaneous frequencies of chromosome and chromatid aberrations and rate of micronucleus (MN) formation in both cell lines. We have also characterized the frequency of sister chromatid exchange (SCE) in transgenic Big Blue mouse cells. Big Blue Rat2 cells are hyperploid and have extremely high baseline frequencies of cytogenetic damage. In addition, Big Blue Rat2 cells are BrdU-resistant, therefore, SCE frequencies cannot be assessed in these cells. We conclude that Big Blue Rat2 cells are not useful for routine cytogenetic toxicology studies. The transgenic Big Blue mouse cell line is polyploid and consistently yields a low mitotic index (approximately 1%) in untreated cells. These mouse cells also exhibited moderately high baseline frequencies of chromosome and chromatid aberrations, however, baseline frequencies of SCE and of MN were not elevated. Transgenic Big Blue mouse embryonic fibroblasts were further studied for MN induction following treatment with N-ethyl-N-nitrosourea (ENU) for 0.5 h at concentrations of 0.425, 0.85 and 1.7 mM. Concentration dependent increases in MN were observed in these cells. Thus, while an ENU induced cytogenetic response using transgenic Big Blue mouse cells demonstrates that this cellular model could be used to cytogenetically complement the mutagenesis assays, the low mitotic index and the high spontaneous frequency of chromosome damage confounds its use for routine genetic toxicology studies. PMID- 9862200 TI - A 2.3 A resolution structure of chymosin complexed with a reduced bond inhibitor shows that the active site beta-hairpin flap is rearranged when compared with the native crystal structure. AB - In the crystal structure of uncomplexed native chymosin, the beta-hairpin at the active site, known as 'the flap', adopts a different conformation from that of other aspartic proteinases. This conformation would prevent the mode of binding of substrates/inhibitors generally found in other aspartic proteinase complexes. We now report the X-ray analysis of chymosin complexed with a reduced bond inhibitor CP-113972 ?(2R,3S)-isopropyl 3-[(L-prolyl-p-iodo-L-phenylalanyl-S methyl-cysteinyl)amino-4]-cyclohexy l-2-hydroxybutanoate? at 2.3 A resolution in a novel crystal form of spacegroup R32. The structure has been refined by restrained least-squares methods to a final R-factor of 0.19 for a total of 11 988 independent reflections in the resolution range 10 to 2.3 A. The extended beta-strand conformation of the inhibitor allows hydrogen bonds within the active site, while its sidechains make both electrostatic and hydrophobic interactions with residues lining the specificity pockets S4-->S1. The flap closes over the active site cleft in a way that closely resembles that of other previously determined aspartic proteinase inhibitor complexes. We conclude that the usual position and conformation of the flap found in other aspartic proteinases is available to native chymosin. The conformation observed in the native crystal form may result from intermolecular interactions between symmetry-related molecules in the crystal lattice. PMID- 9862201 TI - Crystal structure of carboxypeptidase A complexed with an inactivator in two crystal forms. AB - Two different crystal forms of carboxypeptidase A (CPA) complexed with an inactivator were obtained by the method of hanging drop vapor diffusion. The inactivator, 2-benzyl-3-iodo-propanoic acid (BIPA), binds covalently to an active site residue Glu270 of CPA. The complexes were crystallized in the space group P2(1) (CPA-I) and P2(1)2(1)2(1) (CPA-II), respectively. The structures of both crystal forms were determined by molecular replacement using the native CPA crystal structure as the search model. The final crystallographic residuals are 0.163 for CPA-I and 0.152 for CPA-II. Except for the modification of Glu270, the inactivator exhibits normal binding mode compared with other ligand complexes of CPA. In the final electron density difference maps (2Fo-Fc, Fo-Fc), the density of the iodo ion could not be found in both crystal forms while the conserved water molecule remains coordinated to Zn2+ as in the native CPA. Comparisons of the complexes of CPA-BIPA with the native CPA and the CPA-D-Phe complex are presented. The mechanism of the inactivation of CPA and its implication for catalytic mechanism were discussed. PMID- 9862202 TI - NMR analysis of the N-terminal SRCR domain of human CD5: engineering of a glycoprotein for superior characteristics in NMR experiments. AB - CD5 is a type-I transmembrane glycoprotein found on thymocytes, T-cells and a subset of B-cells. The extracellular region consists of three domains belonging to the scavenger receptor cysteine-rich (SRCR) superfamily for which the three dimensional polypeptide fold is as yet unknown. Glycosylated CD5 domain 1 (CD5d1) has been obtained by expression by secretion from both Chinese hamster ovary (CHO) cells and Pichia pastoris. Recombinant CD5d1 expressed in this manner was shown to be correctly folded by binding to anti-CD5 L17F12/Leu1 monoclonal antibody. Preliminary nuclear magnetic resonance (NMR) spectra obtained for CD5d1 (residues 1-118) had spectral dispersion typical of a folded protein, but otherwise of such poor quality that NMR structural studies were not feasible. The analysis of glycoproteins by NMR is frustrated by sample heterogeneity and poor spectral quality associated with glycan resonance overlap and the potential for increased line-widths due to the large hydrodynamic volume. In order to pursue NMR structural studies of CD5d1 it was necessary to optimize the quality of NMR spectra of CD5d1. A range of constructs of varying length and carbohydrate content were expressed in CHO cells and in P. pastoris. In addition the P. pastoris CD5d1 proved susceptible to N-glycan cleavage with endoglycosidase H. The protein products were characterised using size exclusion chromatography, NMR measurement of translational self-diffusion coefficients and two-dimensional 1H nuclear Overhauser effect spectroscopy experiments. Removal of an eight residue O glycosylated C-terminal peptide, in particular, resulted in significant improvements in the quality of the CD5d1 NMR data, while retaining native protein structure. Two-dimensional heteronuclear NMR spectroscopy of nitrogen-15 isotope labelled deglycosylated CD5d1 (residues 1-110) prepared from P. pastoris suggests that this protein product is now amenable to solution structure determination. PMID- 9862203 TI - Sequences annotated by structure: a tool to facilitate the use of structural information in sequence analysis. AB - With the aim of bridging the gap between protein sequence and structural analyses, we have developed a tool to aid the identification of new protein sequences by recognizing distant homologues using structural information. The tool generates sequence annotated by structure (SAS) files, applying structural information derived from structural analyses to a given protein sequence. A World Wide Web interface allows a given sequence to be submitted either for structural annotation or, where its structure is unknown, for search and alignment against sequences of known structure. In both cases, SAS will colour residues in the sequence of known structure according to a selection of properties, including secondary structure, interatomic contacts and active site information. SAS can also be used to inspect properties of a single structure. PMID- 9862204 TI - In unison: regularization of protein secondary structure predictions that makes use of multiple sequence alignments. AB - We present a method whose purpose is to post-process the fuzzy results of secondary structure prediction methods that use multiple sequence alignments, in order to obtain 'realistic' secondary structures, i.e., secondary structure elements whose length is greater than or equal to some predefined minimum length. This regularization helps with interpretation of the secondary structure prediction. PMID- 9862205 TI - Proteins from thermophilic and mesophilic organisms essentially do not differ in packing. AB - The role of the packing density in the elevation of thermal stability of proteins from thermophilic organisms is widely discussed in the literature. In the present study, this issue was reconsidered in the scale of an unbiased set of protein structures. Partial specific volumes, void and cavity volumes were calculated for a set of 80 non-homologous proteins and for 24 proteins from thermophilic organisms and analysed in the context of their possible role in thermal stabilization. The results showed that there is no significant difference between the two sets in respect to the partial specific volume and cavity volume. The proteins from thermophilic organisms showed a slight tendency of increasing void volume, i.e. reducing the packing density. However this observation was not confirmed by the comparison of this parameter for proteins within different structural families. The results suggested that neither the reduction of the packing density nor the reduction of the packing defects can be considered as a common mechanism for increasing the thermal stability of the proteins from thermophilic organisms. Combining the result from this and our previous study we concluded that the electrostatic interactions seem to be a common factor regulating the thermal tolerance of proteins from thermostable organisms. PMID- 9862206 TI - Concerted motions in the photoactive yellow protein. AB - Molecular dynamics simulations have been performed with the aim of identifying concerted backbone motions in the photoactive yellow protein. Application of the essential dynamics method revealed large, chromophore-linked fluctuations of the protein in the ground state, as well as in a form containing the isomerized chromophore. Various loops become more mobile upon isomerization of the chromophore, including a loop which is part of the PAS domain motif, found in light perception proteins. The hinge points identified in these fluctuations correlate with the positions of evolutionary conserved glycines. The results derived from the simulations directly correlate with available experimental data, provide a framework for understanding the dynamic behaviour of the yellow protein and give clues to subsequent steps in the signal transduction pathway. PMID- 9862207 TI - Effects of counter-ions and volume on the simulated dynamics of solvated proteins. Application to the activation domain of procarboxypeptidase B. AB - Molecular dynamics (MD) simulations of the globular activation domain of porcine procarboxypeptidase B (ADBp) and its isolated alpha-helix 1 were performed in order to understand the effects of adding salts and using periodic boundary conditions (this being reflected in the box size) along the simulations. alpha Helix 1 was chosen because it is the most charged element of the secondary structure within ADBp. Different types of MD simulations with the GROMOS package were performed, studying either the whole activation domain or the isolated alpha helix 1 with different water box sizes and counter-ionic shells. The analyses of the trajectories show that simulations of solvated proteins are highly sensitive to the presence of counter-ions and less sensitive to the volume of the water box. The differences in protein potential energies, r.m.s. deviations and radius of gyration between the simulations with and without counter-ions demonstrate that during such studies secondary structures of proteins are more stable when their charges are carefully neutralized. This stresses the need for such a procedure when analysing significantly charged proteins. The results also showed that the enlargement of the water box helps in the stabilization of the system. PMID- 9862208 TI - Hinge-bending motions in annexins: molecular dynamics and essential dynamics of apo-annexin V and of calcium bound annexin V and I. AB - Annexins are homologous proteins that bind to membranes in a calcium dependent manner, but for which precise physiological roles have yet to be defined. Most annexins are composed of a planar array of four homologous repeats, each containing five alpha-helices and associated into two modules. Annexin V forms a voltage-gated calcium channel in phospholipid bilayers. It has been proposed that the hydrophilic pore in the centre of the molecule may represent the ion conduction pathway and that a hinge movement in annexin V causes a variation of the inter-module angle and opens the calcium ion path. Here we present the results of molecular dynamics simulations of apo-annexin V and of calcium-bound annexin V and annexin I. The three simulations show significant differences in conformation and dynamics. The essential dynamics method was used to study the essential subspace of annexin V and showed that one of the essential motions corresponds to the postulated hinge motion. The hinge residues were located between repeats but belong to helices rather than to the links between helices. Calcium binding to annexin V led to a limitation of this hinge motion with more open conformations being favoured. PMID- 9862209 TI - Computational site-directed mutagenesis of haloalkane dehalogenase in position 172. AB - The application of molecular modelling and quantum-chemistry calculations for the 'computational site-directed mutagenesis' of haloalkane dehalogenase is described here. The exhaustive set of single point mutants of haloalkane dehalogenase in position 172 was constructed by homology modelling. The ability of substituting residues to stabilize the halide ion formed during the dehalogenation reaction in the enzyme active site was probed by quantum-chemical calculations. A simplified modelling procedure was adopted to obtain informative results on the potential activity of mutant proteins in a sufficiently short period of time, which, in the future, could be applicable for making bona fide predictions of mutants' activity prior to their preparation in the laboratory. The reaction pathways for the carbon-halide bond cleavage were calculated using microscopic models of wild type and mutant proteins. The theoretical parameters derived from the calculation, i.e. relative energies and selected atomic charges of educt, product and transition state structures, were statistically correlated with experimentally determined activities. The charge difference of educt and product on the halide stabilizing hydrogen atom of residue 172 was the best parameter to distinguish protein variants with high activity from mutant proteins displaying a low activity. All mutants with significant activity in the experiment were found to have this parameter one order of magnitude higher than mutants with low activity. The results obtained are discussed in the light of the practical application of this methodology for the prediction of potentially active protein variants. Further automation of the modelling procedure is suggested for combinatorial screening of the large number of protein variants. Coupling of the dehalogenation reaction with hydrogenation of the halide ion formed during the reaction in the enzyme active site was proposed as a possible way to improve the catalytic activity of the haloalkane dehalogenase of Xanthobacter autotrophicus GJ10. PMID- 9862210 TI - Structural modeling of the pro-ocytocin-neurophysin precursor. AB - The hormonal precursor pro-ocytocin-neurophysin is activated by selective cleavage at Arg2-Ala13, producing mature ocytocin and neurophysin. To understand the cleavage mechanism better, and in particular the recognition of the cleavage site, it is necessary to characterize the three-dimensional structure of the precursor molecule. Here we combine a variety of experimental data with molecular modeling and dynamics calculations to derive possible precursor conformations. In the models obtained, the N-terminus of the precursor, corresponding to the ocytocin segment, is hydrogen bonded in a pocket of the neurophysin moiety in a similar manner to a crystallographically obtained non-covalent complex between the two molecules. The calculations suggest that although the ocytocin segment is relatively flexible, it adopts a stable, broad loop structure in the vicinity of the cleavage region, which may constitute the structural element recognized by the cleaving enzyme. The calculations also suggest a possible widening of the distance between the two neurophysin domains in the precursor relative to that in the non-covalent neurophysin-ocytocin complex. PMID- 9862211 TI - Metal chelating properties of adenylate kinase from Paracoccus denitrificans. AB - Zinc, a common element of adenylate kinases from Gram-positive bacteria, binds to a structural motif consisting of three or four cysteine residues, Cys-X2-Cys-X16 Cys-X2-Cys/Asp. The enzyme from Paracoccus denitrificans, a Gram-negative bacterium, has structural features much similar to those of adenylate kinases from Gram-positive organisms [Spurgin, P., Tomasselli, A.G., and Schiltz, E. (1989) Eur. J. Biochem., 179, 621-628]. However, adenylate kinase isolated from this bacterium was not reported to bind metal. These findings prompted us to clone the corresponding gene of P. denitrificans, and to characterize the enzyme overproduced in Escherichia coli. The deduced primary structure of adenylate kinase from P. denitrificans revealed two differences from that previously published: Cys was found at position 130 instead of His, and His was found at position 138 instead of Gly. The recombinant enzyme is a dimer which binds either zinc or iron, in a metal/monomer ratio of one. The dissociating sulfhydryl reagent, p-(hydroxy-mercuri)phenylsulfonate, released the metal from the protein, confirming that thiols are involved in zinc- or iron-binding. The iron-chelated form of recombinant P. denitrificans adenylate kinase, which is essentially under reduced form, transfers electrons to the oxidized cytochrome c. In conclusion, the absence of metal in the enzyme isolated from P. denitrificans is not related to the protein structure but most probably due to the physiological properties of the host organism. PMID- 9862212 TI - Decreasing the stability and changing the substrate specificity of the Bacillus stearothermophilus alcohol dehydrogenase by single amino acid replacements. AB - The gene encoding the alcohol dehydrogenase (adh-hT) from the thermophilic bacterium Bacillus stearothermophilus LLD-R strain has been overexpressed in Escherichia coli and the corresponding recombinant protein purified to homogeneity. Two putative structural determinants contributing to the higher stability of ADH-hT had been identified by comparison with the less thermostable ADH (ADH-T) from the less thermophilic B. stearothermophilus NCA 1503. In order to ascertain their role, mutations were designed to eliminate in ADH-hT a salt bridge at the N-terminus and a proline residue in the coenzyme binding domain replacing the amino acids located at the same positions in ADH-T. Three mutants- Glu11Lys, Pro242Ala, and Glu11Lys/Pro242Ala--were expressed at high level and the proteins purified and characterized. In general, the mutations had little effect on the activity, indicating that they were not disruptive. The thermal resistance was changed displaying quite additive effects. PMID- 9862213 TI - Protein engineering of BamHI restriction endonuclease: replacement of Cys54 by Ala enhances catalytic activity. AB - Chemical modification studies of BamHI endonuclease indicated the importance of the cysteine residue in catalysis [Nath, K. (1981) Arch. Biochem. Biophys, 212, 611-617]. Of the three cysteine residues at positions 34, 54 and 64 in the BamHI endonuclease Cys54 and Cys64 are at the DNA-protein interface. The co-crystal structure of the BamHI-DNA complex, however, does not indicate any role of cysteines either in binding or catalysis. In the context of strong biochemical evidence, Cys54 in BamHI was changed to Ala54 to investigate its role in catalysis. The mutation was carried out by PCR overlap extension, the mutant gene was cloned and characterized by sequencing. The mutant BamHI was expressed and purified to homogeneity and the kinetic parameters (K(M) and kcat) of the wild type and the C54A mutant were determined. The mutation results in up to approximately 40% enhancement of kcat and some increase in K(M). These in vitro results were also supported by in vivo SOS induction assays: the C54A mutant gene under the T7 promoter caused complete lysis in JH139 in absence of T7 RNA polymerase whereas the wild-type gene gave deep blue colonies under the same conditions. The results suggest no direct role of Cys54 in catalysis, but it can influence the catalytic activity through Val57 backbone contact seen in the co crystal structure. PMID- 9862214 TI - Identification of two new hydrophobic residues on basic fibroblast growth factor important for fibroblast growth factor receptor binding. AB - Basic fibroblast growth factor (bFGF) is implicated in the pathogenesis of several types of vascular and connective diseases. A key step in the discovery of bFGF receptor antagonists to mitigate these actions is to define the functional epitopes required for receptor binding of the growth factor. Using structure based site-directed mutagenesis, two critical areas on the bFGF surface for the high affinity receptor binding have already been identified [Springer, B.A., Pantoliano, M.W., Barberal, F.A., Gunyuzlu, P.L., Thompson, L.D., Herblin, W.F., Rosenfeld, S.A. and Book, G.W. (1994) J. Biol. Chem., 269, 26879-26884; Zhu, H.Y., Ramnarayan, K., Anchin, J., Miao, Y., Sereno, A., Millman, L., Zheng, J., Balaji, V.N. and Wolff, M.E. (1995) J. Biol. Chem., 270, 21869-21874; Zhu, H.Y., Anchin, J., Ramnarayan, K., Zheng, J., Kawai, T., Mong, S. and Wolff, M.E. (1997) Protein Engng, 10, 417-421]. According to these studies, one receptor binding site includes two polar residues Glu96 and Asn104 on bFGF whereas the other includes four hydrophobic residues Tyr24, Tyr103, Leu140 and Met142. Using a protein modelling technique, we report here the identification of a new hydrophobic patch on bFGF which includes residues Tyr73, Val88 and Phe93. The role of this area on receptor binding affinity was evaluated by mutating each of these residues individually and determining the mutated protein's (mutein's) receptor binding affinity. In addition, we examined the role of two other hydrophobic residues, Phe30 and Leu138, on bFGF for high-affinity receptor binding. These two residues are the neighbors of the hydrophobic residues Tyr24 and Tyr103, respectively. Replacement of Val88 and Phe93 with alanine reduced the receptor binding affinity about 10- and 80-fold, respectively, compared with wild type bFGF. In contrast, substitution of Phe30 and Leu138 with alanine has no effect on the receptor binding affinities. We conclude that the newly identified hydrophobic residues, Val88 and Phe93, are crucial for the receptor binding. The present data, together with the previous identification of four hydrophobic residues (Tyr24, Tyr103, Leu140 and Met142), suggests that there are two hydrophobic receptor binding sites on the bFGF surface. Our findings can be employed in the discovery and design of potent bFGF antagonists using computational methods. PMID- 9862215 TI - Stepwise transformation of a cholera toxin and a p24 (HIV-1) epitope into D peptide analogs. AB - We have transformed two peptide epitopes into D-peptide analogs: VPGSQHIDS derived from cholera toxin recognized by the antibody TE33, and GATPQDLNTML from the HIV-1 capsid protein p24 recognized by the antibody CB4-1. The transformation process was performed by stepwise substitution of each single epitope position by all 19 D-amino acids and glycine followed by antibody binding studies and selection of one D-analog for further transformation. Thus, each transformation step introduced one novel D-position into the peptide. For both epitopes complete D-analogs were obtained. The cholera toxin-derived variant dwGsqhydp binds to the antibody TE33 with higher affinity than its original epitope, whereas in the case of the p24-derived analog saGdwwGkssl lower affinity was detected. Both D peptides are completely stable in serum for several days. Antibody interaction models for both D-molecules were generated by computer-assisted modelling based on the crystal structures of the starting complexes. Compared with the L peptides, the binding conformation of dwGsqhydp is very similar, whereas saGdwwGkssl displays a completely different interaction mode. PMID- 9862216 TI - Prediction of epitopes and production of monoclonal antibodies against gastric H,K-ATPase. AB - Monoclonal antibodies (mAbs) were produced against gastric H,K-ATPase using a theoretical and experimental strategy based on prediction of linear epitopes by molecular modelling followed by production of anti-peptide antibodies. By analysing the alpha subunit sequence, we predicted several epitopes corresponding to amino acids K519-L533, E543-Y553 and S786-L798 and produced monoclonal antibodies HK519, HK543 and HK786. All three react against gastric H,K-ATPase in RaLISA, immunohistochemistry and Western blots demonstrating that they recognize the native and the SDS-denatured ionic pump and that the epitopes are located at the surface of the native ATPase. Antibody Kd are in the range 6-10x10(-8) M. Monoclonal antibody HK519 is a competitive inhibitor of ATP, in agreement with ATP binding to K519. Neither mAb 543, nor mAb 786 inhibit the ATPase activity. Monoclonal antibody 95111, whose epitope is mapped between residues C529 and E561, competes with mAb HK543 but not with the other two. We suggest that the 95111 epitope is overlapping or very close to the HK543-553 sequence. Induction of E1 conformer by binding FITC to K519 increases the number of mAb 95111 and mAb HK543 epitopes but not that of mAb 786, supporting the fact that the fragment E543-Y553 changes accessibility, maybe during the E1-E2 transconformation. PMID- 9862217 TI - Distribution of hyaluronan and dermatan/chondroitin sulfate proteoglycans in human aortic dissection. AB - Aortic dissections (AD) are characterized by the separation of the artery into two sheets, possibly due to fragility of the vessel wall. A mucoid histological pattern, imparted to the tissues mainly by hyaluronan and proteoglycans, can be seen in "cysts" and, in chronic cases, in a band of repair tissue. We studied the localization of hyaluronan, versican, decorin and biglycan in situ in aortas of 21 patients with recent AD, 8 with chronic AD and in 15 control cases. None of these substances was increased in the areas of mucoid "cysts" that possibly contain anomalous material. Similar distributions were seen in normal and dissected aortas: versican and hyaluronan were more prominent in the external half of the medial layer where the dissection usually occurs. Since these molecules play a role in resistance to compression, disorders not detected by our method may be involved in aortic dissection. Hyaluronan was seen adjacent to fibrin at the dissection tear, probably as an early wound repair phenomenon. Biglycan, hyaluronan and mostly versican are seen during advanced repairing. The mucoid deposits may represent various compounds which reflect different disorders in vascular biology. PMID- 9862218 TI - Cell death and proliferation and its relation to collagen degradation in uterine involution of rat. AB - Collagen concentration, procollagenase localization, and their association with cell proliferation and apoptosis during postpartum involution, were investigated biochemically and histochemically in postpartum day 1, 3, 5, and 7 rat uterine tissues. In control animals, uterine wet weight, soluble protein, and collagen decreased rapidly during days 1 to 3 postpartum, and the DNA concentration in the uterine horn rapidly decreased, as noted by others. Simultaneously, both apoptosis and cell proliferation were observed in these tissues. These processes were highest in smooth muscle cells on day 3 postpartum. Procollagenase was found in the cell cytoplasm through days 1 to 3 postpartum, was highest on the third day postpartum, and appeared to gradually diminish by day 5 postpartum. Disorganization of collagen fibers was observed, under polarized microscopy by a strong birefringence of collagen fibers of the circular smooth muscle cell layers. However, this disorganization of the uterine collagen diminished progressively from day 3 to day 7. Treatment with estradiol or a combination of estradiol and progesterone suppressed cellular turnover and attenuated the changes in DNA, total amino acids, and collagen on day 3 postpartum. In this study, cellular turnover and biochemical and morphological changes appeared to be closely associated. Gonadal steroid hormones appear to influence these changes and retard uterine involution. This study suggests that a dynamic turnover of the cellular population takes place during uterine involution. It is possible that other factors, in addition to steroid hormones, contribute to uterine involution. It is to be postulated that these factors either are themselves decreased or, alternatively, may increase the inhibition of other unknown factors by an indirect mechanism. PMID- 9862219 TI - Rat osteoclast precursors in vivo express a vitronectin receptor and a chloride bicarbonate exchanger. AB - In vivo osteoclast precursors, which are mononuclear, were previously found to express TRAP (tartrate-resistant acid phosphatase) and CTR (calcitonin receptor), like multinucleated osteoclasts. In vitro, they were found to express, in addition, VNR (vitronectin receptor) and CBE (chloride-bicarbonate exchanger). In order to ascertain that osteoclast precursors in vivo express VNR and CBE like their in vitro counterparts, we used immunohistochemistry to localize these molecules in developing long bones of neonatal rats. Frozen sections of metatarsals and phalanges of 1-2 day-old rats were stained for TRAP and mineralization using histochemistry or were reacted with polyclonal antibodies specific for either the beta3 chain of the VNR or synthetic sequences of the CBE. Both mature, multinucleated osteoclasts within the forming marrow cavity of metatarsals (as shown previously) and mononuclear osteoclast precursors located outside the bony collar of the phalangeal calcified rudiment (as shown here for the first time) expressed both TRAP, VNR and CBE. These findings suggest that mononuclear osteoclast precursors express many of the phenotypical markers of multinucleated osteoclasts prior to their fusion and multinucleation which may allow them to resorb bone, as suggested by in vitro observations of pit formation by preosteoclasts cultured on resorbable substances. PMID- 9862220 TI - Banded patterns in liquid crystalline phases of type I collagen: relationship with crimp morphology in connective tissue architecture. AB - Solutions of type I acid soluble collagen were studied in light and electron microscopy at concentrations over 40 mg/ml. Banded patterns spontaneously emerge in samples observed between crossed polars between slide and coverslip. The textures are interpreted as precholesteric, appearing at the transition between the isotropic phases, due to random molecular order, and the cholesteric phase corresponding to a highly organized three-dimensional structure. Type I collagen banded patterns correspond to regular undulations of the molecular directions with an observed periodicity in the range of 1 to 10 microm. This interpretation is verified by ultrastructural analysis of precholesteric samples gelled under ammonium vapors. Results are discussed in regard to banded patterns described either within synthetic polymer systems or within collagen extracellular matrices. Self-assembled liquid crystalline phases of collagen generate crimp morphologies. Their possible relationship with early secretion steps in the development of connective tissues is discussed. PMID- 9862221 TI - Localization of carboxy-terminal type II procollagen peptide (pCOL-II-C) and type II collagen in the repair tissue of full-thickness articular cartilage defect. AB - It is well established that a full-thickness articular cartilage defect is repaired with a fibrocartilaginous tissue of which cells are derived from undifferentiated mesenchymal stem cells in the bone marrow. To characterize the repair tissue immunohistochemically, full-thickness defects were created in rabbit knee joints, and the repair tissues immunostained at 3, 6, and 12 weeks after surgery. Well characterized polyclonal antibody against carboxyterminal type II procollagen peptide (pCOL-II-C) and monoclonal antibody against type II collagen were used to evaluate the repair tissue with regard to the metabolism of type II collagen. Immunohistochemistry revealed that pCOL-II-C was localized in or around most of the repair cells obtained at 3 and 6 weeks after surgery, while type II collagen distributed mainly in the pericellular matrix of metaplastic round-shaped repair cells. The results suggest that the repair cells taken at the early stage of the repair process of the defect could originally have more activity of type II collagen synthesis. PMID- 9862222 TI - Comparison of surgically attached and non-attached repair of the rat Achilles tendon-bone interface. Cellular organization and type X collagen expression. AB - The effects of surgical repair versus non-repair on cell morphology and type X collagen expression were investigated using a rat model of Achilles tendon avulsion. The animals were divided into four groups. In Group 1, tendon was reattached to the original attachment site by suturing through a drill hole in the calcaneus; in Group II, tendon was not reattached and a drill hole was not made; in Group III, tendon was not reattached but a drill hole was made; and the animals in Group IV were sham operated. In Group I (tendon reattached), at 2 weeks postoperatively, many hypertrophic chondrocytes appeared at the reattachment site adjacent to bone and type X collagen was detected immunologically both in the cells and in the extracellular matrix. After 4 weeks, the cells at the original site of attachment were arranged in rows along the newly formed tendon fibers and were stained with type X collagen antibody. By contrast, when tendon was not reattached (Groups II and III), a gap between the original attachment site and the tendon stump was observed through the entire postoperative period. At 8 weeks, the original attachment site was covered by fibrocartilaginous tissue and tendon became attached to the calcaneal fibrocartilage area, which is proximal to the original attachment site. Type X collagen was detected in the cells which were adjacent to bone. In Group IV (sham operation), there were no changes in histology or type X collagen distribution, either at the attachment site or in tendon and bone, compared with the non operated control rats. These results suggest that surgical reattachment of tendon to the original site is important to help reorganize cells during the repair process. Type X collagen was identified immunohistochemically in the cells adjacent to bone in all the groups, suggesting that it may play a role in maintaining distinct areas of calcified and non-calcified fibrocartilage. PMID- 9862223 TI - Heterotopic implantation of mouse bone-marrow cells: an in vivo model allowing analysis of mineral phases during mineralization processes. AB - Heterotopic calcification induced after implantation of bone-marrow cells under the murine kidney capsule was used to study the mineral phases occurring during the mineralization process. Ossicles were found to contain numerous osteoblastic cells that produced an organic matrix closely associated with active hematopoietic tissue. During implantation of bone marrow, needle-shaped microcrystals were progressively deposited on collagen fibers. The mineral formed in the heterotopic calcification consisted mainly of calcium phosphate. The distribution and density of the microcrystals were heterogeneous after 6 weeks of implantation but became homogeneous and well-crystallized after 10 weeks. The Fourier transform infrared microspectroscopy provided important spatial data on the nature of the mineral formed and the changes in the mineral environment. Similarities were noted between young bone (bone callus) and 6-week heterotopic ossicles, and between adult bone and 10- or 12-week heterotopic ossicles. The study demonstrated that murine heterotopic calcification under the renal capsule can be a very useful model for studying bone apatite formation during the mineralization process. PMID- 9862224 TI - Cloned 3T6 cell line from CD-1 mouse fetal molar dental papillae. AB - Only primary pulpal cell cultures and one virally transformed mouse cell culture have been formally reported in the literature to synthesize proteins such as phosphophoryn which are unique to dentin matrix. In the present study, a mixed culture was derived from dental papilla cells of 18-19 fetal day CD-1 mouse mandibular first molars, maintained on a 3T6 plating regimen, and subsequently cloned after 28 passages. This cloned cell line (MDPC-23) exhibited several unique features, some of which were characteristic of odontoblasts in vivo. The features of this cell line included (1) epithelioid morphology of all cells with multiple cell membrane processes, (2) high alkaline phosphatase activity in all cells, (3) formation of multilayered nodules and multilayered cultures when maintained in ascorbic acid and beta-glycerophosphate, and (4) expression of two markers for odontoblast differentiation, i.e. dentin phosphoprotein and dentin sialoprotein. PMID- 9862225 TI - Expression of dentin sialoprotein (DSP) and other molecular determinants by a new cell line from dental papillae, MDPC-23. AB - The purpose of this study was to characterize the molecular expression of a spontaneously immortalized and cloned cell line (MDPC-23) derived from 18-19 day CD-I fetal mouse molar dental papillae to determine if these cells were odontoblast-like. Western blots showed that a protein band, at approximately 105 kDa, reacting positively with anti-DSP antibodies and co-migrating with mouse DSP, was present in lysates of cells from passages 7, 37 and 77, in serum-free conditioned medium from passage 37 cells, and in mouse dentin extract. A minor band at 55 kDa was also apparent in cell lysates. Using a cDNA probe for a 486bp mouse DSP coding sequence, DSP or DSP-PP mRNA expression was detected by Northern analysis as well as Southern analysis after RT-PCR in all three passages. It was also shown that in these cells 1,25 (OH)2 vitamin D3 upregulated both osteopontin and osteocalcin mRNA, and dexamethasone downregulated alkaline phosphatase and alpha2(I) collagen mRNA. Thus, MDPC-23 cells express proteins which are common to mineralizing tissue. The expression of DSP and DSP-PP strongly suggests that this cell line is from the odontoblast lineage. PMID- 9862226 TI - Signaling pathway by which TGF-beta1 increases expression of latent TGF-beta binding protein-2 at the transcriptional level. AB - The cytokine transforming growth factor-beta has multiple effects on a wide variety of cell types. These effects include modulation of growth and regulation of gene transcription. In the present work, we demonstrate that TGF-beta1 increases transcription of the latent transforming growth factor-beta binding protein-2 ( LTBP-2) gene in cultured human fetal lung fibroblasts leading to a significant increase in LTBP-2 mRNA steady state level. The stability of LTBP-2 mRNA was not appreciably altered. A corresponding increase in production of LTBP 2 protein accompanied the increase in mRNA. Through the use of specific inhibitors, we demonstrate that a member of the Ras super family and a protein kinase C, probably of the atypical (non-diacylglycerol, non-Ca++ dependent) class are likely to be components in the signaling pathway. However, phospholipases, G proteins and extracellular-signal regulated kinases do not appear to be involved. These results combined with previous findings on elastin regulation by TGF-beta1 (Kucich et al. (1997). Am. J. Respir. Cell Mol. Biol., 17: 10-16) demonstrate that TGF-beta1 can coordinately increase the steady state levels of mRNAs encoding components of the elastic fiber, but through diverse mechanisms. In contrast to LTBP-2, increased elastin expression is achieved by message stabilization. Furthermore, the TGF-beta1 signaling pathways differ and while the pathway leading to increased LTBP-2 transcription shares components with those modulating transcription of other genes, it is unlikely to be precisely congruent with any other previously described one. PMID- 9862227 TI - Identification of distinct metabolic pools of aggrecan and their relationship to type VI collagen in the chondrons of mature bovine articular cartilage explants. AB - The metabolism and distribution of newly synthesized aggrecan present in the extracellular matrix of intact explant cultures of mature articular cartilage was investigated with respect to type VI collagen-stained chondrons. Using biochemical, autoradiographical and novel confocal immunohistochemical techniques it was shown that aggrecan exists as a number of distinct pools that are located within the extracellular matrix of the tissue. The first was identified as a pool of high specific radioactivity, much of which appeared in the medium one day after incubation with radiolabeled sulfate. Of the radiolabeled aggrecan remaining within the extracellular matrix, three pools were differentiated on the basis of time and location within the extracellular matrix. One pool was resident within the pericellular microenvironment associated with the chondron, one migrated into the territorial matrix adjacent to the chondron and one was sequestered long term in the interterritorial matrix. Analysis of the kinetics of loss of radiolabeled aggrecan macromolecules present in the region of matrix defined by the chondron suggests that this pool rapidly turns over and is a precursor to the pools of aggrecan present in the territorial and interterritorial matrix. There were marked differences in the distribution of newly synthesized aggrecan present in these regions of the extracellular matrix in explant cultures maintained with or without fetal calf serum. In the absence of serum, more of the newly synthesized aggrecan moved into the territorial and interterritorial matrix indicating that the presence of serum in the culture medium influenced the tissue distribution of aggrecan. This work indicates that the pericellular microenvironment of the chondron plays an important role in the retention and maturation of aggrecan prior to the sequestration of aggrecan complexes into the functional load bearing matrices of adult articular cartilage. PMID- 9862228 TI - Increase of decorin content in articular cartilage following running. AB - The effect of long distance running exercise (40 km/day for 15 weeks, five days a week) on the decorin content of articular cartilage from the knee joint was studied in female beagle dogs. Samples from load bearing sites on the lateral plateau of the tibia (TL), and pooled material from two minimum load bearing sites on the posterior section of lateral (FLP) and medial (FMP) femoral condyles were analyzed. The running exercise protocol did not lead to significant changes in the overall glycosaminoglycan content of the cartilage. However, the amount of decorin significantly increased in the TL samples, and also in the FMP pool. These results support earlier in vitro observations that decorin synthesis is stimulated by loading, independent of the synthesis of aggrecan. PMID- 9862229 TI - Acylated ascorbate stimulates collagen synthesis in cultured human foreskin fibroblasts at lower doses than does ascorbic acid. AB - Acylated derivatives of ascorbic acid were found to be active in a number of biochemical and physiological processes. In the present study we investigated the effects of 6-O-palmitoyl ascorbate on collagen synthesis by cultured foreskin human fibroblasts. Our observations indicate a marked stimulatory effect on collagen synthesis by 6-O-palmitoyl ascorbate in the concentration range of 5-20 microM, while the synthesis stimulated by ascorbic acid was maximal at concentrations of 20-100 microM. Cells treated with 10 microM palmitoyl ascorbate for 36 h exhibited a production of collagen threefold greater than those in the presence of 10 microM ascorbic acid, and it was about the same as in cells treated with 100 microM ascorbic acid. By 48 h differences were not significant. Acylated ascorbate impaired vitality of the treated fibroblasts at concentrations exceeding 20 microM in media supplemented with 0.5% FCS. However, most of the cytotoxic effect was neutralized by FCS at a concentration of 10%. The resistance of acylated ascorbate against oxidative degradation as well as the role of free radicals in the modulation of collagen synthesis by ascorbic acid and by its derivatives is discussed. PMID- 9862230 TI - Ontogenesis of hepatocyte respiration processes in relation to rat liver cytochrome P450-dependent monooxygenase system. AB - The study aimed to evaluate the effect of age on the activity of the cytochrome P450-dependent monooxygenase system and on cellular respiration processes in Wistar rats aged 0.5, 1, 2, 4, 8, 12, 20, and 28 months. The following parameters were determined: cytochrome P450 content, cytochrome b5 content, NADPH-cytochrome P450 reductase activity, and NADH-cytochrome b5 reductase activity, succinate dehydrogenase (SDH) activity, and lactate dehydrogenase (LDH) activity. In the study, cytochrome P450 content increased in the first month of life, which was accompanied by increases in SDH and LDH activities. In the subsequent months, SDH activity decreased, whereas LDH activity increased to reach the maximum in month eight and then decreased. Cytochrome b5 content showed a decreasing tendency throughout the experiment. NADH-cytochrome b5 reductase activity showed only slight deviations in individual age groups. PMID- 9862231 TI - Modification of HLA expression on peripheral lymphocytes and monocytes during aging. AB - Immunosenescence involves modifications of humoral and cellular immunity. Here we report the analysis of human leukocyte antigen (HLA) expression on T lymphocytes, B lymphocytes and monocytes of 58 healthy subjects aged 23-95 years old. Using a double staining immunofluorescence and flow cytometry analysis, we have determined the percentages of cells expressing HLA class-I and HLA-DR antigens. The number of antigenic sites expressed per cell were evaluated for HLA-ABCw, HLA A, HLA-B, HLA-DR locus with a flow cytometry quantification technique. With advancing age, we observed: (i) a significant decrease of the percentage of T cells and B cells expressing HLA-A products; (ii) a decrease of the number of HLA class-I antigenic sites expressed per cell on the three populations tested, predominantly on B cells and in a locus-dependent fashion; (iii) a decrease of the number of HLA-DR molecules expressed per T cell, although the percentage of T cells expressing DR products was increased; (iv) a significant diminution of the percentage of B cells expressing HLA-DR molecules, without changes of the number of HLA-DR antigenic sites per cells. These changes in HLA expression with increasing age could contribute to the decreased level of immunologic responsiveness observed with ageing and contribute to the modification of antigen recognition. PMID- 9862232 TI - Cell-associated pentosidine as a marker of aging in human diploid cells in vitro and in vivo. AB - Cellular aging is characterized by alterations at both the morphological and molecular levels, some of which are decreased mitotic rate, increased cytoplasmic vacuolization, and changes in intrinsic cellular constituents (Stanulis-Praeger, 1987. Mech. Ageing Dev. 38, 1-48). In the present investigation, glycoxidation is studied as a marker for cellular aging by measuring cell-associated pentosidine levels in human skin fibroblasts as a function of replicative life span and in human peripheral blood T lymphocytes as a function of chronological age. Fibroblasts were isolated from culture by detachment/centrifugation while lymphocytes were isolated from blood by a Ficoll-Paque/Lympho-Kwik T-Cell Prep technique. Pentosidine levels were measured in acid-hydrolyzed cell pellet suspensions by high-pressure liquid chromatography. Results show that pentosidine was detected in early and late cultured reticular and papillary fibroblasts. Pentosidine, expressed as either protein, DNA, or cell number, significantly (P < 0.0006) increased with in vitro passage and was significantly (P < 0.01) related to cell proliferation as measured by cell density and cell doublings per day during culture. Cell-associated pentosidine was measured in T lymphocytes isolated from healthy, diabetic, and uremic individuals. In healthy controls, levels significantly (P < 0.0003) increased with age. In uremic individuals, a large variation was observed with many values above the 95% confidence intervals determined for controls. Since a previous study showed that plasma pentosidine in healthy subjects does not increase with age, these results suggest that cellular turnover perhaps coupled to a deterioration in cellular anti-glycoxidation defensive mechanisms play a substantial role in explaining increased pentosidine concentrations during cellular aging. PMID- 9862233 TI - Dynamic phenotypic restructuring of the CD4 and CD8 T-cell subsets with age in healthy humans: a compartmental model analysis. AB - In healthy humans, phenotypic restructuring occurs with age within the CD3+ T lymphocyte complement. This is characterized by a non-linear decrease of the percentage of 'naive' (CD45RA+) cells and a corresponding non-linear increase of the percentage of 'memory' (CD45R0+) cells among both the CD4+ and CD8+ T-cell subsets. We devised a simple compartmental model to study the age-dependent kinetics of phenotypic restructuring. We also derived differential equations whose parameters determined yearly gains minus losses of the percentage and absolute numbers of circulating naive cells, yearly gains minus losses of the percentage and absolute numbers of circulating memory cells, and the yearly rate of conversion of naive to memory cells. Solutions of these evaluative differential equations demonstrate the following: (1) the memory cell complement 'resides' within its compartment for a longer time than the naive cell complement within its compartment for both CD4 and CD8 cells; (2) the average, annual 'turnover rate' is the same for CD4 and CD8 naive cells. In contrast, the average, annual 'turnover rate' for memory CD8 cells is 1.5 times that of memory CD4 cells; (3) the average, annual conversion rate of CD4 naive cells to memory cells is twice that of the CD8 conversion rate; (4) a transition in dynamic restructuring occurs during the third decade of life that is due to these differences in turnover and conversion rates, between and from naive to memory cells. PMID- 9862234 TI - Ribosomal RNA in Alzheimer's disease and aging. AB - Ribosomal RNA genes are involved in cell transcription and translation processes and can modulate gene expression. In an earlier cytogenetic study, (Payao, S.L.M., Smith, M.de A.C., Kormann-Bortolotto, M.H., Toniolo, J., 1994 (Investigation of the nucleolar organizer regions in Alzheimer's disease. Gerontology 40, 13-17), reported a decreased activity of ribosomal genes in Alzheimer's disease (AD). We studied the ratio of mature rRNA 28S and 18S in peripheral blood samples derived from eight patients with AD, eight healthy elderly sisters of these patients (SA), eight healthy elderly (EC) and eight healthy young (YC) controls, all female. Our results showed a statistically significant decrease of mature rRNA 28S and 18S ratio in the elderly groups (AD, SA, EC) in relation to the young one, probably by fragmentation of 28S rRNA. The Alzheimer's patient group had the lowest 28S/18S ratio. Thus, we can suggest that there is a possible change in the transcriptional or maturation process, or a preferential degradation of the 28S subunit with ageing. PMID- 9862235 TI - Calorie restriction, stress and the ubiquitin-dependent pathway in mouse livers. AB - Calorie restriction (R) is the only known method to delay the aging process and extend mean and maximal lifespan in rodents. R has been shown to delay the age related accumulation of damaged proteins and to protect organisms from various stresses which can produce damaged proteins. Such stresses include irradiation, heat shock, and oxidative stress. The ubiquitin- and ATP-dependent proteolytic pathway (UPP) has been associated with the degradation of abnormal and/or damaged proteins. We examined the effect of diet and oxidative stress on activities of the UPP in supernatants from livers taken from 23-month-old Emory mice which had been exposed to an in-vivo injection of paraquat. Paraquat induces oxidative stress by generating superoxide radicals. In livers from non-stressed animals, steady-state levels of endogenous ubiquitin conjugates, de novo conjugate formation, and E1 and E2 activities were significantly lower in R animals than in control (C) animals. However, after exposure to paraquat, levels of endogenous ubiquitin conjugates were significantly higher in R versus C animals, and de novo conjugate formation and E1 and E2 activities in R animals rose to levels which were indistinguishable from levels of these activities noted in C animals. R was associated with an increased ability to degrade beta-lactoglobulin by the UPP after an oxidative stress was imposed. Ability to degrade beta-lactoglobulin by the C or R livers in non-stressed animals was not significantly different. Taken together, these data indicate that oxidative stress in R animals is associated with enhanced levels of ubiquitin conjugates and that this enhancement may be due to an increase in UPP activity. These data also indicate that the ability to form ubiquitin conjugates and the UPP system does not change with oxidative stress in C animals. The latter is consistent with prior reports that suggests that older C animals may already be in a state of enhanced oxidative stress and that activities of the UPP provide a sensitive indicator of levels of cellular redox status. PMID- 9862237 TI - The fractionation experiment: reducing heterogeneity to investigate age-specific mortality in Drosophila. AB - Age-specific mortality rates decelerate at older ages in both genetically homogenous and heterogeneous populations of Drosophila. One explanation proposed for deceleration is population heterogeneity. This hypothesis suggests that a population consists of sub-populations that differ in mortality characteristics and that the deceleration is the result of selective survival of stronger individuals. Here we describe an experiment that fractionates populations into several sub-populations without changing the physiological characteristics of the post-fractionated populations. Through a careful process of selection of Drosophila eggs, larvae, pupae and adults, we attempt to reduce as much as possible the degree of pre-adult, environmentally induced heterogeneity among individuals of a genetically identical cohort. We then ask whether such cohorts, when compared to non-fractionated populations, exhibit a lesser degree of mortality deceleration at advanced ages. From a total of 106 fractionated and control populations, consisting of 51331 individuals, 101 populations (93% of the fractionated populations and 100% of the control populations) exhibit a significant amount of mortality deceleration late in life. These observations suggest that environmental heterogeneity accrued during larval development is not a major factor contributing to mortality deceleration at older ages. PMID- 9862236 TI - Decrease in Ca-ATPase activity in aged synaptosomal membranes is not associated with changes in fatty acyl chain dynamics. AB - We have examined lipid peroxidation (LPO) and fatty acid acyl chain dynamics in synaptosomal membranes isolated from aged rat (Fischer 344 x Brown Norway F1 hybrids) brains, correlating these results with measurements of enzymatic activity of the synaptic plasma membrane Ca2(+)-ATPase (PMCA). Calcium-dependent ATPase activity in these membranes exhibits progressive decreases with a maximal loss of activity with age of approximately 35%. The sensitivity of this membrane bound ion transporter to the lipid composition of the surrounding membrane, as well as the high abundance of oxidatively sensitive polyunsaturated fatty acyl chains in synaptosomal membranes, suggests that this age-related loss in catalytic turnover may result from LPO-mediated protein modification and/or changes in the physical structure of the bilayer. However, high-performance liquid chromatography analysis of 2,4-dinitrophenylhydrazone derivatives reveals no significant age-related increases in the content of reactive aldehydes (malondialdehyde, formaldehyde, acetaldehyde or acetone) which comprise breakdown products of lipid peroxidation. Electron paramagnetic resonance measurements employing 5- and 12-stearic acid spin labels with the nitroxide reporter groups at two depths in the bilayer were used to assess the fatty acyl chain dynamics (fluidity) of synaptosomal membranes. The resulting spectra demonstrate anisotropic lipid dynamics of two populations of lipids, i.e. lipids in direct association with membrane proteins (boundary lipids) and bulk lipids that do not directly associate with proteins. The nanosecond dynamics of both lipid populations is unaltered with age indicating that any compositional changes occurring with age are insufficient to result in alterations in bilayer fluidity relevant to PMCA activity. Thus, the observed age-related decline in PMCA activity may be explained by direct modification of membrane protein. PMID- 9862238 TI - Mortality from congenital anomalies. PMID- 9862240 TI - Prescribing for the elderly patient: why do we need to exercise caution? PMID- 9862239 TI - Safety assessment of peroxide antimalarials: clinical and chemical perspectives. PMID- 9862241 TI - The lignocaine metabolite (MEGX) liver function test and P-450 induction in humans. AB - AIMS: The N-deethylation of lignocaine to monoethylglycinexylidide (MEGX) is partially catalysed by the rifampicin inducible P-450 isoenzyme CYP3A4. This has led to the use of the MEGX test (MEGX plasma concentrations after i.v. lignocaine) as a marker of CYP3A4 activity. To test this hypothesis, we studied lignocaine and MEGX plasma pharmacokinetics. METHODS: Ten healthy volunteers received rifampicin (600 mg day(-1)) for 6 days, resulting in a four- to sixfold increase in urinary 6beta-hydroxycortisol output. On days 1 and 7 (pretreatment), day 11 (treatment), and day 14 (48 h after rifampicin), 50 mg lignocaine i.v. was administered. MEGX concentrations at 30 min [MEGX30min] were assessed and normalised to MEGX test results after 1 mg kg(-1) lignocaine. On days 7 and 14 the lignocaine and MEGX plasma concentrations were measured over a 300 min period. MEGX test results and lignocaine and MEGX plasma pharmacokinetics before and after induction with rifampicin were compared. RESULTS: The lignocaine plasma clearance increased from 7.5+/-1.2 ml min(-1) kg(-1) before to 8.6+/-2ml min(-1) kg(-1) (P=0.026) after induction. The normalised MEGX30min concentrations increased from 61+/-14 (day 7) to 82+/-34 microg l(-1) (day 14) by a mean of 21 microg l(-1) (95% confidence interval: -3 to 44 microg l(-1)) (P=0.055). CONCLUSION: An insignificant increase of MEGX plasma concentrations was found in 10 volunteers after induction of CYP3A4 activity by rifampicin. Therefore, the MEGX test is not a sensitive marker of P-450 induction in healthy human liver. PMID- 9862242 TI - Influence of urine pH and urinary flow on the renal excretion of memantine. AB - AIMS: The present study assessed the influence of urinary flow rate and urine pH on the renal excretion of the NMDA-receptor antagonist memantine. METHODS: In a randomized, open, four-period cross-over trial, 12 healthy male volunteers received 10 mg memantine daily for 43 days. After reaching steady state conditions the volunteers were allocated to four different regimens to alter urine pH and urinary flow, which were each separated by a 1 week period while the study medication continued (A: acidification of urine pH, low urinary flow; B: acidification of urine pH, high urinary flow; C: alkalinization of urine pH, low urinary flow; D: alkalinization of urine pH, high urinary flow). RESULTS: The renal clearance of memantine (CL(R)) in regimen A and B was 7-10 fold higher in comparison with regimen C and D (P<0.05). There were small but statistically significant differences of CL(R) between the two regimens with acidic urine pH (A: median: 210.2 ml min(-1) vs B: median: 218.7 ml min(-1)) and between the two regimens with alkaline urine pH (C: median: 19.4 ml min(-1) vs D: median: 30.5 ml min(-1)). The amount of memantine excreted into the urine within one regimen (Ae0 24h) was 5.7-7.4 fold higher in regimens A and B than C and D (P< 0.05). Differences of the AUC(0,24 h) and Cmax/AUC(0,24 h) were significant (P<0.05) between each of the regimens with acidic urine pH (A, B) and regimens (C, D) with alkaline urine pH (A vs C, A vs D, B vs C, B vs D) but not between regimens A vs B or C vs D. CONCLUSIONS: The present study demonstrated a considerable effect of urine pH, whereas no clinically relevant change of the renal excretion of memantine with urinary flow could be detected. As the renal excretion of memantine may have an impact on therapeutic efficacy changes of dietary habits that may alter urine pH should be avoided during treatment with memantine. PMID- 9862243 TI - Lamotrigine and therapeutic drug monitoring: retrospective survey following the introduction of a routine service. AB - AIMS: To review (retrospectively) the relationships between lamotrigine (LTG) dosage and plasma concentrations based on data generated in a routine therapeutic drug monitoring laboratory from a heterogeneous sample of patients with epilepsy. To distinguish patients taking concomitant anti-epileptic therapy which induced or inhibited drug metabolising enzymes, or a combination of both, together with LTG. To survey medical staff who use a routine LTG assay service with a view to establishing the utility of higher plasma LTG concentrations than those used in early clinical trials. METHODS: All patient assays for LTG received over a 12 month period (339 requests from 149 patients) were reviewed and relationships between dosage and concentration calculated and grouped according to concomitant antiepileptic drug therapy. The doctors requesting the tests were surveyed by questionnaire (n=40 of 67 responded). They were asked for details about the patient's seizure control, rationale used for LTG dosage adjustment and their acceptance of the proposed 'therapeutic range' adopted by the laboratory of 3-14 mg(-1). RESULTS: Linear relationships were demonstrated between LTG dosage and concentration for the 3 treatment groups (LTG plus valproic acid (VPA), LTG plus enzyme inducing antiepileptic drugs, and LTG plus VPA and inducers), however, there were significant differences between groups (P<0.001) with a 4.4 fold difference in dosage: concentration ratios between the LTG plus VPA group and the LTG plus inducers group. The questionnaire showed that the therapeutic range was well accepted by 88% of responders, none of whom considered this higher range to be wrong. CONCLUSIONS: Metabolic inhibition by VPA was shown to have a marked effect on LTG kinetics, suggesting either a significant LTG dosage reduction is required if plasma LTG concentrations are elevated, or alternatively, higher plasma LTG concentrations could be attained from lower dosages. The higher therapeutic range adopted by the laboratory (3-14 mg(-1)) was widely accepted and increasingly applied in clinical practice in the management of patients with epilepsy. PMID- 9862244 TI - Population pharmacokinetics and therapeutic response of CGP 56697 (artemether + benflumetol) in malaria patients. AB - AIMS: To investigate the pharmacokinetic and pharmacodynamic properties of artemether and benflumetol in a fixed combination tablet (CGP 56697) and to offer an explanation for the lower than expected cure rate in a Thai clinical trial. METHODS: Two hundred and sixty patients were enrolled into a randomized, double blind, parallel group, dose-finding trial. CGP 56697 was given orally, either as: A, 4 x 4 tablets over 48 h; B, 4 x 2 tablets over 48 h or C, 3 x 4 tablets over 24 h. Each tablet contained artemether 20 mg amd benflumetol 120 mg. The pharmacokinetics were determined using a population-based approach combining full profiles (42 patients) and sparse data (218 patients). Parasite clearance time and 28 day cure rate were correlated with the derived pharmacokinetic parameters. RESULTS: The median absorption half-life of benflumetol was 5.3 h, with a tmax of 10 h and terminal elimination half-life of 4.5 days. For artemether (and its metabolite, dihydroartemisinin), the corresponding values were 1.9 (1.9) h, 1.8 (1.2) h, and 0.84 (0.43) h. The variability in bioavailability of artemether and dihydroartemisinin was large both between doses and between patients, but was less pronounced for benflumetol. Compared with the first dose, benflumetol bioavailability was estimated to increase three-fold by the third and fourth doses. Higher artemether or dihydroartemisinin AUC was found to decrease parasite clearance time. Higher benflumetol AUC was found to significantly increase the chance of cure. CONCLUSIONS: Using a population-based approach it was confirmed that the pharmacokinetic and pharmacodynamic properties of benflumetol and artemether differ markedly. Benflumetol AUC is associated with cure and the effect of benflumetol when coadministered with artemether is to prevent recrudescence. The mode of action of benflumetol is consistent with its longer elimination half-life. A short course of low-dose artemether, which is rapidly absorbed and has a short elimination half-life, produced effective parasite clearance. The complementary pharmacokinetic and pharmacodynamic properties of benflumetol and artemether was the main rationale for developing a fixed-dose combination. While the 4 x 4 dose regimen is very effective in most endemic areas, the poorer absorption (2.5 fold lower than in China) and the more resistant parasites in Thailand require higher doses of this drug. PMID- 9862245 TI - Ribavirin and interferon alfa-2b in chronic hepatitis C: assessment of possible pharmacokinetic and pharmacodynamic interactions. AB - AIMS: The primary objective of this study was to determine whether pharmacokinetic interactions occurred between interferon alpha-2b (IFN) and ribavirin in patients with chronic hepatitis C infections. Additionally this study assessed the single and multiple-dose pharmacokinetics of ribavirin and IFN, and compared the safety, tolerability and antiviral pharmacodynamics of IFN plus ribavirin compared with either drug alone. METHODS: In this open label parallel group study, patients with chronic hepatitis C were randomized to receive IFN 3 million IU thrice weekly s.c. alone, ribavirin 600 mg twice daily p.o. alone or both drugs in combination over 6 weeks. Single and multiple dose pharmacokinetics and indices of antiviral pharmacodynamics were assessed during weeks 1 and 6, along with safety assessments during the study. RESULTS: The range of mean ribavirin terminal phase half-lives after single doses was 44-49 h. Comparison of week 1 and week 6 AUC(0,12h) values showed accumulation in plasma of approximately 6-fold. The range of mean washout half-lives after week 6 was 274-298 h, reflecting release of ribavirin from deep compartment stores. The range of single and multiple dose IFN terminal phase half-lives was 5-7 h. IFN demonstrated an increase in bioavailability (approximately 2-fold) upon multiple dose administration. Ribavirin and IFN pharmacokinetic parameters for combined ribavirin and IFN were similar to those during monotherapy with either compound, although the power of this study to detect differences was low. Serum HCV-RNA titers and ALT concentrations were reduced by IFN alone, ribavirin alone reduced ALT concentrations only, and combined IFN plus ribavirin produced numerically greater falls in both measurements than either treatment alone. Serum concentrations of neopterin and activity of 2',5'-oligoadenylate synthetase (2'5' OAS) were increased by IFN alone and in combination with ribavirin, whereas serum 2'5'-OAS activity was decreased and neopterin concentrations unaltered by ribavirin monotherapy. IFN and ribavirin monotherapy produced characteristic changes in safety laboratory tests (IFN--reductions in white cells, neutrophils and platelets; ribavirin--reduced haemoglobin) and characteristic adverse event profiles (IFN--headache, flu-like symptoms, fatigue, anorexia, nausea, myalgia, and insomnia; ribavirin--headache, fatigue, myalgia, and pruritus). There was no additive effect of combination therapy on safety laboratory tests or reported adverse events. All changes were fully reversible upon treatment cessation. CONCLUSIONS: There was no evidence of pharmacokinetic interactions between IFN and ribavirin in this study. There were numerical trends indicating that the combination of IFN and ribavirin reduced titers of HCV-RNA to a greater extent than did either treatment alone, and the safety profile of combination therapy was similar to those of both monotherapy treatments. PMID- 9862246 TI - Frusemide inhibits angiotensin II-induced contraction on human vascular smooth muscle. AB - AIMS: Frusemide is widely used in the treatment of acute pulmonary oedema, chronic congestive heart failure and, to a lesser degree, in the treatment of hypertension. Evidence suggests that frusenuide exerts an anti-vasoconstrictor effect independent of its diuretic properties. Since angiotensin II is a highly potent vasoconstrictor involved in the pathophysiology of these diseases, we have investigated the effect of frusemide on the contraction elicited by angiotensin II on human internal mammary artery (IMA) and saphenous vein (SV). METHODS: Rings of IMA and SV were suspended for isometric tension recording in organ baths. Concentration-response curves to angiotensin II were performed in the absence (control) or in the presence of frusemide (10(-6) to 10(-3) M). In addition, the effect of frusemide was evaluated after cyclooxygenase inhibition by indomethacin (10(-6) M) and was compared with those of the other loop diuretic bumetanide (10( 4) M). RESULTS: Frusemide induced a concentration-dependent decrease of the contraction elicited by angiotensin II on IMA and SV. On both vessels, the inhibitory effect on the maximal contraction to angiotensin II was significant with concentrations of frusemide from 10(-5) to 10(-3) M. Angiotensin II potency (pD2) was only reduced by 10(-3) M frusemide. The effect of frusemide was not altered in the presence of indomethacin. Bumetanide was less potent than frusemide in inhibiting angiotensin II-induced contractions in both IMA and SV. CONCLUSIONS: Frusemide, at concentrations in the therapeutic range (10(-5) M), inhibits angiotensin II-induced contraction on human isolated IMA and SV. This inhibitory effect is cyclooxygenase independent and appears mediated, at least in part, by inhibition of Na+/K+/2Cl- symport. Reduction in the vasoconstrictor effect of angiotensin 1 may be involved in the therapeutic efficacy of frusemide. PMID- 9862247 TI - Comparison of the vasoconstrictor effects of rizatriptan and sumatriptan in human isolated cranial arteries: immunohistological demonstration of the involvement of 5-HT1B-receptors. AB - AIMS: We compared the vasoconstrictor effects of 5-HT with those of the selective 5-HT1B/1D-receptor agonists sumatriptan and rizatriptan in human isolated cranial (middle meningeal) arteries. In addition selective 5-HT1B- or 5-HT1D-receptor antibodies were used in combination with semiquantitative immunohistochemical techniques to compare the levels of expression of these receptors in human middle meningeal and coronary arteries. METHODS: Middle meningeal and coronary arteries were obtained (with consent) from either neurosurgical patients or donor hearts, respectively. Segments of middle meningeal artery were mounted in organ baths for isometric recording and cumulative concentration-effect curves to 5-HT, rizatriptan and sumatriptan were obtained. Frozen fresh sections of middle meningeal and coronary arteries were subjected to standard immunohistochemical techniques using specific 5-HT1B- or 5-HT1D-receptor primary antibodies and a radiolabelled secondary antibody. Data were subjected to analysis of variance (ANOVA) and nonlinear regression analysis. RESULTS: 5-HT, rizatriptan and sumatriptan were potent vasoconstrictors in human isolated middle meningeal artery (EC50 values=32, 90 and 71 nM, respectively). A significantly higher level of 5-HT1B-receptor immunoreactivity was detected in middle meningeal artery compared with coronary artery (ANOVA, F=7.95, DF=1,4, P<0.05). CONCLUSIONS: Rizatriptan and sumatriptan act selectively to cause vasoconstriction in human isolated middle meningeal artery and are 10-fold more potent than in human coronary artery. The higher level of expression of 5-HT1B-receptors in middle meningeal compared with coronary artery provides a pharmacological basis for the craniovascular selectively of both rizatriptan and sumatriptan. PMID- 9862248 TI - Endogenous angiotensin II and baroreceptor dysfunction: a comparative study of losartan and enalapril in man. AB - AIMS: To assess the role of direct ATI receptor antagonism in baroreceptor modulation in man, and to perform a direct comparison of Ang II blockade at the receptor level with that of ACE inhibition. METHODS: Ten healthy male volunteers [mean age (s.d.) 23 (6.9)] pretreated with frusemide therapy (40 mg day(-1) for 3 days prior to each visit) were studied on 3 separate days, 10 days apart, in a placebo-controlled, randomized, double-blind, cross-over fashion. On each study day, subjects were randomly given either a single-dose of enalapril 20 mg, losartan 50 mg or placebo. Baroreceptor function was assessed by measuring changes in blood pressure (BP), pulse interval (RR Int) and heart rate (HR) in response to incremental doses of intravenous phenylephrine infusions (0.2-3.6 microg kg(-1) min(-1)). RESULTS: In response to phenylephrine, no significant differences in BP responses were observed with any of the study medications but reflex heart rate responses were significantly increased with both enalapril and losartan compared with placebo (P<0.05). The (RR/AsBP ratio, taken as a measure of baroreceptor sensitivity (BRS) was significantly increased with enalapril 112.2+4.6 ms mmHg (mean+s.d.)] and losartan [11.9+3.6msmmHg(-1)] compared with placebo [9.2+4.5 ms mmHg(-1)]; i.e. enalapril and losartan increased the (RR/(delta sBP ratio by 3.0 ms mmHg(-1) (95%CI 0.5, 5.6; P<0.05) and 2.8 ms mmHg( 1) (95%CI 0.6, 5.0; P< 0.038), respectively. There were however, no significant differences between losartan and enalapril [mean difference 0.25 (95%CI - 1.6, 2.1)]. CONCLUSIONS: The present study confirms observations from animal models that blocking endogenous angiotensin II in man improves baroreceptor function. Both strategies, ATI receptor antagonism and ACE inhibition appear to be equally effective in restoring baroreceptor function in salt-depleted normotensive subjects. PMID- 9862249 TI - Impaired pressor sensitivity to noradrenaline in septic shock patients with and without impaired adrenal function reserve. AB - AIMS: To investigate the relationship between adrenal gland function and pressor response to noradrenaline in septic shock. METHODS: Basal cortisol level, noradrenaline--mean arterial pressure dose-response curve and cortisol response to intravenous corticotrophin bolus were obtained in nine patients fulfilling usual criteria for septic shock and in six normal volunteers. In patients with septic shock, dose-response curve to noradrenaline was determined a second time 60 min after a 50 mg intravenous hydrocortisone bolus. RESULTS: As compared with controls, patients with septic shock had increased basal cortisol levels (mean+/ s.d.: 1564+/-818 vs 378+/-104 nmol l(-1) , P=0.002, 95% confidence interval for difference in means: [452, 1920]) and a blunted cortisol response to corticotrophin (403+/-461 vs 1132+/-195 nmol l(-1), P=0.008, [-1163, -2951). Five patients had impaired adrenal function reserve. As compared with controls, septic patients displayed a moderate and non significant decrease in pressor sensitivity to noradrenaline (P=0.112). As compared with patients with adequate adrenal response, patients with impaired adrenal function reserve showed a significant decrease in pressor sensitivity to noradrenaline (P=0.038). In septic patients, hydrocortisone improved pressor response to noradrenaline (P=0.032). This effect was more marked in patients with impaired adrenal function reserve so that, as compared with patients with adequate response, the difference was no longer significant (P=0.123). CONCLUSIONS: In septic shock, impaired adrenal function reserve may partly be accounted for by the depressed pressor sensitivity to noradrenaline. The latter may be substantially improved by physiological doses of hydrocortisone. PMID- 9862251 TI - Determination of inulin clearance by bolus intravenous injection in healthy subjects and ascitic patients: equivalence of systemic and renal clearances as glomerular filtration markers. AB - AIMS: Determination of systemic inulin clearance by the standard technique of constant intravenous infusion has long been accepted as a reliable method for measuring glomerular filtration rate (GFR) without urine collection, except in oedematous patients. However, recent studies using standard clearance techniques have claimed that systemic inulin clearance is significantly greater than renal clearance and therefore overestimates GFR. The main purpose of this investigation was to re-evaluate the relationship between systemic and renal inulin clearance using a different technical approach. A reassessment was also made of inulin disposition kinetics. METHODS: Systemic and renal inulin clearances were simultaneously evaluated, in healthy subjects and patients with oedema and ascites, by analysis of the total area under the plasma concentration-time curve (AUC) following bolus intravenous injection. kenal clearance was calculated as the ratio of the total amount recovered in the urine to the AUC, and systemic clearance as dose/AUC. RESULTS: Inulin disposition kinetics were best described by a tri-exponential model. In healthy subjects the volume of the central compartment (mean (s.d.) value 3.86 (1.00) 70 kg(-1)) was slightly greater than the plasma volume; steady-state volume of distribution was 11.00 (1.21) 170 kg( 1), in accordance with the tenet that the inulin space is somewhat smaller than the extracellular fluid volume. The values of systemic and renal inulin clearances were very similar (96.1 (10.0) and 94.6 (12.5) ml min(-1) 70 kg(-1), respectively, in healthy subjects; 104.6 (16.3) and 102.6 (18.5) ml min(-1) in ascitic patients). They were also highly correlated to each other in both healthy subjects (r=0.96, P<0.001) and patients with ascites (r=0.98, P 0.05). PMID- 9862279 TI - Endogenous formaldehyde as a potential factor of vulnerability of atherosclerosis: involvement of semicarbazide-sensitive amine oxidase-mediated methylamine turnover. AB - The mouse is known to be highly resistant to atherosclerosis. However, some inbred mouse strains are vulnerable to atherosclerosis when they are fed a high cholesterol, high-fat diet. Increased deamination of methylamine (MA) and the subsequent production of formaldehyde has been recently shown to be a potential risk factor of atherosclerosis. In the present study semicarbazide-sensitive amine oxidase (SSAO)-mediated MA turnover in C57BL/6 mouse, a strain very susceptible to atherosclerosis, has been assessed in comparison to a moderate, i.e. BALB/c, and resistant, i.e. CD1, mouse strains. Kidney and aorta SSAO activities were found to be significantly increased in C57BL/6 in comparison to BALB/c and CD1 mice. A significant increase of urinary MA and formaldehyde were detected in C57BL/6. [14C]MA following intravenous injection would be quickly metabolized by SSAO. The labeled formaldehyde product would cross link with proteins. C57BL/6 exhibits significantly higher labeled protein adducts than BALB/c and CD1 in response to [14C]MA. The results indicated that mice vulnerable to atherosclerosis possess an increased SSAO-mediated MA turnover. The increase of production of formaldehyde, possibly other aldehydes, may induce endothelial injury or be chronically involved in protein cross-linking and subsequent angiopathy. PMID- 9862280 TI - Prevention of focal intimal hyperplasia in rat vein grafts by using a tissue engineering approach. AB - The present study focused on the role of blood flow in the formation of focal intimal hyperplasia in vein grafts, as well as the development of an engineering approach that can be used to eliminate disturbed blood flow and prevent blood flow-related focal intimal hyperplasia. A rat vein graft model was constructed by interposing a jugular vein into the abdominal aorta with end-to-end anastomoses. Locally disturbed flow was identified by analyzing particle streak-lines in methyl salicylate-cleared and perfused vein grafts in vitro with a physiological Reynolds number. At day 10, 20, and 30 after surgery, focal intimal hyperplasia of the vein grafts was examined using a histological approach and the density of alpha-actin positive cells was determined using immunohistological and fluorescent approaches. Results showed that apparent eddy blood flow formed at the proximal, but not at the distal, end of the vein grafts due to graft-host diameter mismatch and local geometric distortions, and was associated with apparent focal intimal hyperplasia. The thickness of the alpha-actin positive layers of the proximal vein grafts was significantly higher than that of the distal grafts (192 +/- 27 vs. 94 +/- 18 microm, 278 +/- 55 vs. 124 +/- 20 microm, and 288 +/- 24 vs. 131 +/- 23 microm for day 10, 20. and 30, respectively). The density of the alpha-actin positive cells, however, was similar between the proximal and the distal regions (3569 +/- 361 vs. 3285 +/- 343 cells/mm2, 5540 +/ 650 vs. 5376 + 887 cells/mm2, and 5465 +/- 791 vs. 5278 +/- 524 cells/mm2 for day 10, 20, and 30, respectively). When eddy blood flow was eliminated by matching the graft-host diameters using a tissue engineering approach, the average thickness of the alpha-actin positive layers of the proximal (71 +/- 15, 86 +/- 16, and 85 +/- 14 microm for day 10, 20, and 30, respectively) and the distal vein grafts (68 +/- 13, 80 +/- 14, and 79 +/- 13 microm for day 10, 20, and 30, respectively) was reduced significantly. The density of the alpha-actin positive cells was also reduced significantly in the proximal (2946 +/- 359, 3261 +/- 295, 3472 +/- 599 cells/mm2 for day 10, 20, and 30, respectively) and in the distal regions (3151 +/- 511, 3466 +/- 687, 3593 +/- 688 cells/mm2 for day 10, 20, and 30, respectively). The thickness of the alpha-actin positive layers and the density of the alpha-actin positive cells were not significantly different between the proximal and distal regions of the engineered vein grafts at each observation time. These results suggest that eddy flow may develop in vein grafts and may facilitate the formation of focal intimal hyperplasia, and the vascular tissue engineering approach developed in this study may be used to prevent blood flow-related focal intimal hyperplasia in vein grafts. PMID- 9862281 TI - Biochemical determinants of apoptosis and necrosis. AB - Although apoptosis and necrosis were originally thought to be entirely distinct mechanisms of cell death, recent work has shown that the processes are regulated by many of the same biochemical intermediates, most notably the levels of cellular ATP, Ca2+, reactive oxygen species, and thiol antioxidants. Beyond a certain threshold, it appears that stress-induced changes in these modulators 'switches' the cell death mechanism from apoptosis to necrosis. Importantly, even when this occurs, cell death can be attenuated by bcl-2 and caspase inhibitors, which are known for their abilities to block apoptosis. This review will summarize these observations within the context of what is currently known about the effector machinery for apoptotic cell death, and possible mechanistic explanations for the switch between apoptosis and necrosis will be provided. PMID- 9862282 TI - Effect of soman on N-methyl-D-aspartate-stimulated [3H]norepinephrine release from rat cortical slices. AB - Effects of soman on N-methyl-D-aspartate (NMDA) evoked [3H]norepinephrine (NE) release were examined in rat brain cortical slices. NMDA increased [3H]NE release in a concentration-dependent manner. Soman could inhibit the increase evoked by NMDA, but carbachol, an agonist of cholinergic receptor, could potentiate the increase evoked by NMDA. Atropine (a selective muscarinic antagonist) attenuated the release of [3H]NE induced by NMDA in the presence of carbachol or acetylcholine (ACh), but had no effect on the release of [3H]NE induced by NMDA alone. Both d-tubocurarine (an antagonist of nicotinic receptor) and atropine had no effect on the release of [3H]NE induced by NMDA in the presence of soman. These results suggested that soman has a direct action at non-cholinergic sites, probably at NMDA receptors. PMID- 9862284 TI - Determination of perfluorocarboxylic acids by gas-liquid chromatography in rat tissues. AB - A method for the determination of tissue perfluorocarboxylic acids (PFCA) was developed using gas-liquid chromatography with an electron capture detector (ECD). Perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), and perfluorodecanoic acid (PFDA) were efficiently extracted from rat liver, methylated with diazomethane, and separated on GLC. Internal standards that were added to liver homogenates were used in the quantitative analysis of PFCAs to correct the loss during the extraction and derivatization. The concentration of PFDA in rat liver 24 h after intraperitoneal administration at a dose of 20 mg/kg body weight was 113.9 +/- 11.4 microg/g liver. The value corresponds to the previously reported results that were obtained using [14C]PFDA. PMID- 9862283 TI - Activation of alveolar macrophages and peripheral red blood cells in rats exposed to fibers/particles. AB - The cytotoxic and oxidative responses of crocidolite and chrysotile asbestos fibers and ultrafine titanium dioxide (UF-TiO2) particles were measured in alveolar macrophages (AM) and peripheral red blood cells (RBC) of rat after 30 days with a single intratracheal exposure (5 mg). The following responses were observed one month after fiber/particle instillation: (1) AM population increased; (2) lactate dehydrogenase and acid phosphatase activities in cell free lung lavage fluid increased; (3) substances that react with hydrogen peroxide or thiobarbituric acid were elevated in both AM and peripheral RBC; (4) glutathione peroxidase, glutathione reductase, and catalase were altered in both AM and peripheral RBC; (5) glutathione and ascorbic acid decreased in both AM and peripheral RBC. A significant difference from negative controls was noted in all responses of the two fiber-exposed groups, and in most responses of the UF-TiO2 exposed group. The level of responses to the three test substances suggested a decreasing order of toxicity, with crocidolite > chrysotile > UF-TiO2. PMID- 9862285 TI - A labile sulfur in trisulfide affects cytochrome P-450 dependent lipid peroxidation in rat liver microsomes. AB - The effects of trisulfide derivatives were studied on cytochrome P-450-dependent lipid peroxidation using rat liver microsomal systems. Cytochrome P-450-dependent lipid peroxidation was induced by carbon tetrachloride or tert-butylhydroperoxide and was evident by an increase in thiobarbituric acid-reactive substances (TBA RS) and oxygen consumption. In these cytochrome P-450-dependent lipid peroxidation systems, pretreatment of microsome with trisulfide derivatives (cystine trisulfide and thiocyclam) significantly inhibited TBA-RS formation and oxygen consumption compared with disulfide or thiol analogs (cystine, nereistoxin, or cysteine). The labile sulfur contained in trisulfide disappeared during incubation with liver microsomes. In the CCl4-induced lipid peroxidation system, the cytochrome P-450 level and NAD(P)H-cytochrome P-450 reductase activity were significantly decreased by the addition of trisulfide derivatives. Therefore, in cytochrome P-450-dependent lipid peroxidation system, the potential effects of trisulfide appear to be mediated via enzyme inhibition. These results suggest that pretreatment of the trisulfide derivatives may affect the toxic function of exogenous xenobiotics or drugs, which are reduced by the liver enzyme cytochrome P-450 to radical species. PMID- 9862286 TI - Cellular inactivation induced by a radiopharmaceutical kit: role of stannous chloride. AB - Stannous chloride (SnCl2) has been used in many sectors of human activities such as food manufacturing and in nuclear medicine to produce radiopharmaceuticals labeled with technetium-99m (99mTc). Due to its importance and genotoxic potentiality, we decided to evaluate the biological effect induced by a nuclear medicine kit, which includes SnCl2, in association with glucoheptonic acid (GHA) which is employed for brain and renal scintigraphies. These studies were carried out with the Escherichia coli AB1157 strain and the deoxyribonucleic acid (DNA) plasmid pUC 9.1. The experiments, with different concentrations of SnCl2 and GHA, show an inverse relationship between both agents. When the GHA concentration was increased, the cellular inactivation induced by SnCl2 was reduced, as measured by the number of viable cells. Moreover, GHA protects the DNA molecule against the damage induced by SnCl2. PMID- 9862287 TI - Effects of Cd2+, Pb2+ and CH3Hg+ on high voltage-activated calcium currents in pheochromocytoma (PC12) cells: potency, reversibility, interactions with extracellular Ca2+ and mechanisms of block. AB - Effects of the neurotoxic heavy metals Cd2+, Pb2+ and CH3Hg+ on current carried by Ca2+ ions (I(Ca)) through high-voltage activated Ca2+ channels in nerve growth factor (NGF)-differentiated pheochromocytoma (PC12) cells were examined to characterize possible differences in the mechanism of action of these metals on Ca2+ channel function. Specifically, the potency and reversibility of effect on I(Ca) by each metal was examined, as well as the relationship between extracellular [Ca2+] and potency of block of I(Ca) by Cd2+ and Pb2+. In addition, the effect of each of these metals on Ca2+ channels when applied to the intracellular side of the membrane was also examined. When extracellular solution contained 20, 10 or 5 mM Ca2+, the estimated IC50 values (total metal concentration) for block of I(Ca) were 15, 10, and 6.5 microM for Cd2+ and 7.5, 2.0 and 1.1 microM for Pb2+, respectively. CH3Hg+ (1-10 microM) blocked I(Ca) (20 mM Ca2+) in a time- and concentration-dependent manner. When cells were washed with metal-free solutions, block of I(Ca) by Cd2+ was reversed rapidly, whereas block by Pb2+ was reversed only partially, and block of I(Ca) by CH3Hg+ was not reversed. When Pb2+ and CH3Hg+ treated cells were washed in metal-free solutions containing 50 microM D-penicillamine (DPEN), block of I(Ca) by 10 microM Pb2+ was rapidly and completely reversed, whereas, block of I(Ca) by 5 microM CH3Hg+ was not reversed. Higher concentrations (500 microM) of 2,3-dimercapto-1-propane sulfonic acid (DMPS) did reverse partially the block of I(Ca) by 5 and 10 microM CH3Hg+. When Cd2+, Pb2+ or CH3Hg+ was present in the intracellular solution, Ca2+ channel currents were significantly reduced. These results characterize effects of Cd2+ on Ca2+ channels and demonstrate that Cd2+, Pb2+ and CH3Hg+ differ in their actions on Ca2+ channels. PMID- 9862288 TI - Moderate exercise improves gait stability in disabled elders. AB - BACKGROUND: Decreased muscle strength impedes elders' functional performance in daily activities such as gait. The mechanisms whereby increased strength improves gait are unknown. METHODS: A prospective, blinded, randomized trial of moderate intensity strength exercise was conducted and its impact was measured on functional mobility during gait in 132 functionally limited elders. Lower extremity strength was measured, including hip abductor, hip extensor, and knee extensor strength. Of the 132 subjects, 120 subjects (mean age, 75.1 yrs) completed 6 months of elastic band resistance training at least 3 times a week or served as no-exercise controls. RESULTS: Subjects increased their lower extremity strength in the exercise and control groups, by 17.6% and 7.3% (p < .01), respectively. Gait stability improved significantly more in the exercise group than in the control group (p < .05). Increases in forward gait velocity were not significantly different between groups. Peak mediolateral velocity and base of support improved in the exercise group, but not in the control group. Change in lower extremity strength correlated significantly but weakly with many of the gait variables. CONCLUSIONS: Gait stability, especially mediolateral steadiness, improved in the exercise group but not in the control group. These results show that even moderate strength gains benefit gait performance in elders and thus provide a sound basis for encouraging low-intensity strength training for elders with functional limitations. PMID- 9862289 TI - Long-term outcomes and life satisfaction of adults who had pediatric spinal cord injuries. AB - OBJECTIVE: To determine long-term outcomes and life satisfaction of adults who sustained pediatric spinal cord injury (SCI). DESIGN: Structured interview of adults who were 25 years or older who had pediatric SCI. SETTING: Community. PARTICIPANTS: A convenience sample of 46 patients from a total of 81 patients who received care in an SCI program: 1 refused participation, 4 died, and 30 were lost to follow-up. MAIN OUTCOME MEASURES: A structured questionnaire including physical, psychosocial, and medical information. The Craig Handicap Assessment and Reporting Technique and two measures of life satisfaction were also administered. RESULTS: Participants were 25 to 34 years old, mean 27 years. Thirty-two had tetraplegia and 14 had paraplegia. Thirty-one were men. Mean years of education was 14. Fifty-four percent were employed, 48% lived independently, and 15% were married. Life satisfaction was associated with education, income, satisfaction with employment, and social/recreation opportunities, and was inversely associated with some medical complications. Life satisfaction was not significantly associated with level of injury, age at injury, or duration of injury. CONCLUSIONS: Individuals who had pediatric SCI, much like adult-onset SCI, have the greatest opportunity for a satisfying adult life if rehabilitation emphasizes psychosocial factors such as education, employment, and long-term health management. PMID- 9862290 TI - Immunogenicity of pneumococcal vaccine in persons with spinal cord injury. AB - OBJECTIVE: To determine immunogenicity and optimum timing for administering the 23-valent pneumococcal vaccine after spinal cord injury (SCI). DESIGN: Double blind, randomized, placebo control study. SETTING: SCI unit in a tertiary care medical center and community. PARTICIPANTS: Eighty-seven persons with recent SCI. INTERVENTION: Participants were randomized to receive either placebo or pneumococcal vaccine at 16 to 18 days versus 4 to 6 months postinjury. MAIN OUTCOME MEASURES: Antibody concentrations were measured prior to intervention and 1, 2, and 12 months afterward to evaluate the immune response to five serotypes of Streptococcus pneumoniae. Effects of demographic and injury-related variables on immune response were also evaluated. RESULTS: Timing of vaccination did not influence mean antibody concentrations for any serotype (p > .05). Ninety-five percent of vaccinated persons had twofold or greater increases in antibody concentration for at least one serotype when measured 1 month after vaccination versus 35% of placebo groups (p < .01). After 12 months, 93% of vaccinated persons in both groups maintained antibody concentrations twofold or greater than baseline values. CONCLUSIONS: Most participants developed an immune response to at least one serotype that was maintained for at least 12 months. Immune response varied according to serotype. Given the favorable immune response and no effect of timing, persons with SCI should receive pneumococcal vaccine during initial hospitalization. PMID- 9862291 TI - Electrodiagnostic studies: are they useful in clinical practice? AB - OBJECTIVE: Electrodiagnostic testing (electromyography [EMG] and nerve conduction studies [NCS]) may result in some patient discomfort. The justification for such testing should be based on the expectation that the results will affect patient management. This study was conducted to determine how frequently the results of EMG/NCS change the clinical management of the patient. METHODS: One investigator (MB) spoke to each referring physician after EMG/NCS to determine if any management decisions were altered by the test. RESULTS: One hundred forty consecutive EMG/NCS records were obtained. Follow-up was available on 100 patients. Of 78 patients with abnormal findings on EMG/NCS, 29 (37%) had a diagnosis different from the referring diagnosis. For 43 of the 78 (55%), the physician reported that additional diagnostic testing was undertaken or treatment plans were altered. CONCLUSION: EMG/NCS are useful, informative, and diagnostic in the management of various neurologic disorders. PMID- 9862292 TI - The Capabilities of Upper Extremity instrument: reliability and validity of a measure of functional limitation in tetraplegia. AB - OBJECTIVE: To evaluate the reliability and validity of the Capabilities of Upper Extremity (CUE) instrument, designed to measure upper extremity functional limitations in individuals with tetraplegia. Functional limitations are actions such as reaching or grasping and are a link between the domains of impairment and disability. DESIGN: Survey of people with chronic spinal cord injury. SETTING: Regional spinal cord injury center. SUBJECTS: One hundred fifty-four individuals (140 male) with tetraplegia at least 1 year after injury and followed by the center. Mean age was 36.7 years (SD=11.1). Sixty-eight percent were motor complete. METHODS: The 32-item CUE was administered by telephone interview twice about 2 weeks apart. The motor portion of the Functional Independence Measure (FIM) was collected during the first interview. Upper extremity motor scores and motor levels were obtained from the most recent assessment in the outpatient chart. The instrument was evaluated for internal consistency, reliability, and validity. Exploratory factor analysis was performed to examine scale structure. RESULTS: Homogeneity of the scale was excellent. Cronbach's alpha was .96, and item-total correlations ranged from .49 to .78. Test-retest reliability was high (ICC=.94). All but three items had desired levels of agreement (K > .60). Analysis of variance indicated that the CUE distinguished between motor levels of tetraplegia more than one level apart. The CUE was correlated highly with both motor scores and FIM. Regression analysis indicated that the CUE was better than upper extremity motor scores for predicting FIM scores. The model containing the CUE explained 73% of the variance in FIM and was not enhanced by the addition of motor scores. Factor analysis suggested four potential subscales: arm function (bilateral), right hand function, left hand function, and reaching down. CONCLUSION: The CUE exhibits good homogeneity, reliability, and validity; further work is needed to determine its sensitivity to change in function. PMID- 9862293 TI - Comparing consumer and clinician values for alternative functional states: application of a new feature trade-off consensus building tool. AB - OBJECTIVE: To present the Features-Resource Trade-Off Game (Features Game) as a new method for comparing preferences for alternative outcomes among different groups of people. DESIGN: The Features Game is illustrated by comparing preferences for recovery among the 18 functional status items making up the Functional Independence Measure. Methods involved trading levels of independence (resources) across the different items (features). SETTING: Ten community dwelling consumers with physical disabilities and 10 rehabilitation clinicians participated in four separate expert panels-two in Houston and two in Philadelphia. MAIN OUTCOME MEASURES: Five sets of hierarchical stages defined by the four separate panels specifying the profiles of function believed to most foster independent living. RESULTS: Cognitive and communication skills were selected preferentially over the recovery of physical tasks by all panels, but, in comparison to clinicians, consumers were more willing to accept mild deficits in cognitive skills as trade for realizing earlier recovery of physical abilities. CONCLUSION: The overwhelming choice of cognitive and communication abilities over physical abilities suggests a need to enhance therapeutic efforts in those areas. More subtle differences in consumer and clinician preferences emphasize the importance of establishing consumer-oriented goals. PMID- 9862294 TI - Functional outcome of inpatient rehabilitation in persons with brain tumors. AB - OBJECTIVE: To document functional outcome in persons with brain tumors undergoing inpatient rehabilitation and to compare outcomes with a group of traumatically brain injured patients. DESIGN: Retrospective, descriptive, and case-matched. SETTING: A free-standing inpatient brain injury rehabilitation unit. PARTICIPANTS: Forty consecutive patients with a variety of tumor types (40% were either glioblastoma multiforme or meningioma) and a mean age of 53.1 (SD 15.4) years. Sixty percent were men, 25% had recurrent tumors, and 15% had metastatic disease. Also, 40 patients with traumatic brain injury (TBI) matched for age, gender, and admission functional status. MAIN OUTCOME MEASURES: Change in Functional Independence Measure (FIM) scores, length of rehabilitation stay (LOS), and discharge disposition. RESULTS: The mean LOS for the tumor group was 17.8 (SD 9.9) days, mean FIM gain was 25.4 (SD 20.1) points, and 82.5% were discharged home. No demographic or tumor characteristic was statistically significant in predicting functional outcome at discharge, but greater gains were seen for persons with the diagnosis of meningioma, those with left-sided cerebral lesions, and those not receiving radiation therapy. TBI patients made statistically significant greater gains in total FIM change (34.6 vs 25.4), self care (12.3 vs 8.5), and social cognition (5.2 vs 3.6). However, FIM efficiency and LOS were not statistically different between the TBI and tumor groups (1.9 vs 1.5 FIM points/day and 22.1 vs 17.8 days, respectively). CONCLUSIONS: Daily functional gains made by persons with brain tumor undergoing rehabilitation were similar to those made by a group of persons with TBI matched by age, gender, and admission functional status. Further research should use larger samples and address the impact of psychosocial and team factors on LOS and discharge disposition. PMID- 9862295 TI - Clinical characteristics and functional outcome of stroke patients 75 years old and older. AB - OBJECTIVE: To examine the demographic and clinical characteristics of stroke patients 75 years old and older admitted for rehabilitation, to study medical complications that occurred during rehabilitation, and to document functional outcome and possible factors influencing outcome. DESIGN AND SETTING: A case series of 59 consecutive inpatients admitted to a rehabilitation facility with confirmed strokes over a 2-year period. MEASURES: Patients were selected by rehabilitation physicians for admission into the rehabilitation program. Measures for outcome include the Modified Barthel Index (MBI) and the patient's discharge disposition. RESULTS: The mean age of this cohort was 80.4+/-3.2 years, and mean lengths of stay in the acute and rehabilitation facilities were 15.2 and 37.4 days. Twelve patients had three or more medical illnesses; hypertension and diabetes were the most common. Cognitive impairment, urinary incontinence, and dysphagia requiring tube feeding were present in 45.1%, 33.9%, and 11.9% of patients, respectively. Nineteen patients (32.2%) developed medical complications, and urinary tract infection was the most common. Improvements in functional status, motor power, continence, and dysphagia were noted after rehabilitation. Fifty-three patients (89.8%) were successfully discharged home, 28.8% of whom employed domestic maids as caregivers. The discharge MBI score was strongly predicted by the admission MBI and cognition scores. CONCLUSION: Despite their age, significant functional improvements were documented in this cohort of aged stroke patients, and the majority were discharged home. Admission MBI and cognition scores strongly predicted functional outcome. PMID- 9862296 TI - Ultrasound therapy effect in carpal tunnel syndrome. AB - OBJECTIVE: To investigate the overall effect of repeated ultrasound treatment in carpal tunnel syndrome (CTS). DESIGN: Patient-blinded, placebo-controlled, before after treatment trial. SETTING: University hospital PM&R department outpatient clinic and neurology department electromyography laboratory. PATIENTS: Eighteen women with diagnosis of CTS in 30 hands. INTERVENTIONS: Three groups, each with 10 cases of CTS, were randomly established. Continuous ultrasound therapy, with intensities of 1.5W/cm2 (group A), 0.8W/cm2 (group B), and 0.0W/cm2 (group C), was applied to palmar carpal tunnel area for 5 minutes, 5 days a week, for 2 weeks. OUTCOME MEASURES: Patients were evaluated clinically and electrophysiologically before and after the treatment. RESULTS: At the end of treatment, statistically significant improvement was obtained in clinical parameters in all groups: pain (p < .05), pain/paresthesia at night/day (p < .05), and frequency of awakening at night (p < .05). Although there was no statistically significant before-after difference in electrophysiologic studies, slightly decreased motor nerve conduction velocity and increased motor distal latency were noted in groups A and B, but not in group C. CONCLUSION: Ultrasound therapy in CTS was comparable to placebo ultrasound in providing symptomatic relief, and the probability of a negative effect on motor nerve conduction needs to be considered. PMID- 9862297 TI - The difference in anatomy of the lumbar anterior longitudinal ligament in young African-Americans and Scandinavians. AB - OBJECTIVE: To examine possible structural differences in the lumbar anterior longitudinal ligament between young African-American and Scandinavian people. DESIGN: The lumbar anterior longitudinal ligament was examined in 48 African American and Scandinavian young men and women (17 to 30 years old). Apart from the selection of gender, age, height, and weight, all subjects were recruited consecutively during routine forensic autopsies. Sections of the ligament were obtained from the L1 and the L5 vertebra for thickness and width measurements and histologic preparations. The measurements were performed with a digital vernier caliper with an accuracy of 0.1 mm. RESULTS: The width of the L1 part of the lumbar anterior longitudinal ligament in the African-American group (27.5+/ 1.8mm) was greater than that in the Scandinavian group (17.7+/-1.5mm) (p < .001), and the thickness of the L1 part in the African-American group (3.8+/-0.4mm) was greater than that in the Scandinavian group (3.3+/-0.3mm) (p < .001). The width of the L5 part in the African-American group (34.1+/-2.0mm) was greater than that in the Scandinavian group (25.0+/-1.5mm) (p < .001), and the thickness of the L5 part in the African-American group (3.3+/-0.3mm) was greater than that in the Scandinavian group (2.2+/-0.3mm) (p < .001). The cross-sectional area was greater in the African-American group in both sections (L1, African-American 106.4+/ 13.7mm2 vs Scandinavian 52.8+/-7.9mm2; L5, African-American 112.2+/-14.6mm2 vs Scandinavian 55.6+/-6.7mm2) (p < .001). No histologic differences between the two groups were observed. CONCLUSIONS: The lumbar anterior longitudinal ligament was considerably wider and thicker, which yielded a markedly greater cross-sectional area, in African-American than in Scandinavian subjects. In contrast, the ligament appears to be histologically similar in the two groups. These are previously unrecognized differences in anatomy between young African-American and Scandinavian people. PMID- 9862298 TI - Reflex sympathetic dystrophy: is the immune system involved? AB - OBJECTIVE: Evaluation of immune system function in patients with reflex sympathetic dystrophy (RSD). DESIGN: Survey on blood samples obtained from RSD patients and from a randomly selected control group. The lymphocyte populations (T, B, NK cells), and the activated T cells (CD25, and HLA-Dr-positive CD4 and CD8 cells) were analyzed by flow cytometry with dual-color direct immunofluorescence after whole-blood lysis. Clinical chemistry parameters were analyzed in additional serum samples. SETTING: Tertiary care center (outpatient rehabilitation clinic). SUBJECTS: Thirteen patients (nine women) with RSD and a control group of 21 healthy individuals. MAIN OUTCOME MEASURES: The results of the flow cytometry analysis of RSD patients were related to those of the control subjects. Means were analyzed, and confidence intervals for differences of the means were calculated. The means of the clinical chemical analysis were related to local reference values. RESULTS: The flow cytometry analysis did not differ between RSD patients and healthy controls. Although in some patients an individual parameter of clinical chemical analysis differed from its reference value, all of the mean values were within reference limits. Stratification on medications with immunomodulatory effects and on probability of a definite diagnosis of RSD had no influence on the results. CONCLUSION: No association between immunologic indices and RSD was found. This finding is relevant, because recent theories stress that it is not the sympathetic nervous system but a local inflammatory reaction that is fundamental in the pathogenesis of RSD. The results of this study do not support this theory. PMID- 9862299 TI - Bladder shape impact on the accuracy of ultrasonic estimation of bladder volume. AB - OBJECTIVE: Variations in bladder shape can lead to errors in ultrasonic estimation of the bladder volume. The purposes of this study were to compare the accuracies of various formulas to estimate bladder volume from sonographic measurements and to assess the impact of bladder shape on the accuracy of bladder volume estimation. STUDY DESIGN: Twenty-four healthy volunteers (13 men, 11 women) and 55 spinal cord injury patients (48 men, 7 women) underwent ultrasonographic measurements of dimensions before and after voiding. Bladder shape was classified as cuboid, ellipsoid, or triangular prism. Ten formulas from the literature were applied to estimate bladder volume, using the volume voided or catheterized as the standard, and then linear regression was used to obtain optimal correction coefficients for the whole data set as well as each of the three bladder shapes. SETTING: Rehabilitation hospital affiliated with a medical college. RESULTS: The most accurate of the 10 formulas tested was height (H) X transverse depth (Dt) x weight (W) x 0.7 (mean error 17.4%+/-11.6%). Linear regression analysis yield optimal correction coefficients of .72 for the whole data set and .89, .81, and .66 for cuboidal, ellipsoid, and triangular prism shaped bladders, respectively. The mean error for the estimation of bladder volume using .72 as the correction coefficient was 16.9%+/-11.9% and decreased to 12.7%+/-10.1% (p < .0001, paired t test) when the bladder shape was taken into account. CONCLUSION: Using bladder shape for reference and applying the corresponding correction coefficient to volume calculations will improve the accuracy of the estimation. PMID- 9862300 TI - Sonographic diagnosis of hydronephrosis in patients with spinal cord injury: influence of bladder fullness. AB - OBJECTIVE: To investigate and compare the diagnostic accuracy of prevoid and postvoid renal sonography in detecting hydronephrosis in patients with spinal cord injury. STUDY DESIGN: A prospective, blind comparison of renal sonography and excretory urography in 67 spinal cord injury patients who underwent periodic urologic examinations. Renal sonography was performed twice, once when the patient's bladder was physiologically full (prevoid) and again when it was just emptied (postvoid). RESULTS: Of 140 kidneys, 24 from 16 patients were found to have hydronephrosis by excretory urography; 116 kidneys had normal urogram findings. Prevoid sonography missed the diagnosis of hydronephrosis in one kidney and showed hydronephrosis in 18 kidneys that had normal results on excretory urography (sensitivity, 95.8%; specificity, 84.5%; negative predictive value, 99.0%). The postvoid sonograms did not detect hydronephrosis in four kidneys and showed hydronephrosis in six kidneys that had normal results on excretory urography (sensitivity, 83.3%; specificity, 94.8%; negative predictive value, 96.5%). Compared to excretory urography, renal sonography detected eight more upper urinary tract abnormalities, which were confirmed by cystograms or radioisotopic renograms. CONCLUSION: Performing renal sonography while the bladder is full can increase the sensitivity in detecting hydronephrosis in asymptomatic spinal cord injured patients. PMID- 9862301 TI - Neuropathic pain in Charcot-Marie-Tooth disease. AB - OBJECTIVES: To determine the frequency and extent to which subjects with Charcot Marie-Tooth (CMT) disease report pain and to compare qualities of pain in CMT to other painful neuropathic conditions. STUDY DESIGN: Descriptive, nonexperimental survey, using a previously validated measurement tool, the Neuropathic Pain Scale (NPS). PARTICIPANTS: Participants were recruited from the membership roster of a worldwide CMT support organization. MAIN OUTCOME MEASURES: NPS pain descriptors reported in CMT were compared with those reported by subjects with postherpetic neuralgia (PHN), complex regional pain syndrome, type 1 (CRPS-1), also known as reflex sympathetic dystrophy, diabetic neuropathy (DN), and peripheral nerve injury (PNI). RESULTS: Of 617 CMT subjects (40% response rate), 440 (71%) reported pain. with the most severe pain sites noted as low back (70%), knees (53%), ankles (50%), toes (46%), and feet (44%). Of this group, 171 (39%) reported interruption of activities of daily living by pain; 168 (38%) used non narcotic pain medication and 113 (23%) used narcotics and/or benzodiazepines for pain. The use of pain description was similar for CMT, PHN, CRPS-1, DN, and PNI in terms of intensity and the descriptors hot, dull, and deep. CONCLUSIONS: Neuropathic pain is a significant problem for many people with CMT. The frequency and intensity of pain reported in CMT is comparable in many ways to PHN, CRPS-1, DN. and PNI. Further studies are needed to examine possible pain generators and pharmacologic and rehabilitative modalities to treat pain in CMT. PMID- 9862302 TI - Skin breakdown in children and high-frequency oscillatory ventilation. AB - OBJECTIVE: To investigate the relationship of high-frequency oscillatory ventilation (HFOV) to skin breakdown on the scalp and ears in mechanically ventilated children. STUDY DESIGN: Retrospective cohort study of 32 patients supported with HFOV paired with 32 patients supported with conventional mechanical ventilation (CV) in a pediatric intensive care unit (PICU). RESULTS: By univariate analysis, more HFOV patients had skin breakdown than did the CV patients (53% vs 12.5%, p=.001); HFOV patients also had greater severity of illness (Pediatric Risk of Mortality scores), higher mortality, and longer durations of neuromuscular blockade, low systolic blood pressure, and time exposed to risk. Life table analysis demonstrated no difference in the rate of skin breakdown between HFOV and CV patients. Multifactorial analysis showed that only PICU time at risk was a risk factor for skin breakdown. CONCLUSIONS: HFOV was not an independent risk factor for the development of skin breakdown. PICU time at risk was the sole risk factor for the development of skin breakdown in all mechanically ventilated patients in the PICU. PMID- 9862303 TI - Obstacle course performance and risk of falling in community-dwelling elderly persons. AB - OBJECTIVE: To determine the efficacy of obstacle course performance scores in predicting persons at risk for falls, after adjusting for age, sociodemographic, health-status, and physiologic variables. DESIGN: Correlational descriptive study design utilizing a logistic regression model. SETTING: Community setting. PATIENTS OR PARTICIPANTS: A convenience sample of 352 community-dwelling elderly individuals at sites in a metropolitan area. MAIN CRITERION MEASURE: Number of falls reported prospectively during a 2-year follow-up period. RESULTS: Obstacle course performance, fall history, symptoms of balance dysfunction, and activity level distinguished those who fell and those who did not 12 and 18 months later. At 24 months, range of motion and number of medications also were significant. In multivariate logistic regression, only history of a fall was a significant predictor of future falling at 12 and 18 months; at 24 months, the presence of balance dysfunction symptoms was the significant predictor of those who fell. CONCLUSIONS: The obstacle course, as a predictor of future falls, is not superior to the question of whether or not an individual has fallen in the previous year, and is not recommended to predict future falls. The obstacle course may be better as a short-term indicator of response to a rehabilitation program for balance and mobility. PMID- 9862304 TI - Postural changes with aging in tetraplegia: effects on life satisfaction and pain. AB - OBJECTIVES: (1) To measure kyphosis and scoliosis in individuals with tetraplegia; (2) to examine the relation between kyphosis and scoliosis and years since injury; and (3) to determine the association between kyphosis and scoliosis and measures of pain, depression, and life satisfaction. DESIGN: Cross-sectional, case-control study. SETTING: University medical center and a free-standing university-affiliated rehabilitation hospital. PARTICIPANTS: (1) Ten individuals with tetraplegia I to 3 years postinjury (NT); (2) 10 individuals with tetraplegia 10 to 20 years postinjury (OT); and (3) 10 control individuals (C) matched to the other subjects on the basis of age, height, and weight. MAIN OUTCOME MEASURES: Radiographic measurements of kyphosis and scoliosis taken in a seated position, pain as measured by the short form of the McGill Pain Questionnaire (SF-MPQ), depression as measured by the Center for Epidemiological Studies-Depression Scale (CES-D), and life satisfaction as measured by the Life Satisfaction Index Assessment (LSIA) and the Craig Handicap Assessment and Reporting Technique (CHART). RESULTS: No significant differences were seen between the OT and NT groups with respect to age, height, or weight. In addition, no significant differences were found between the NT and OT groups with respect to measures of kyphosis and scoliosis. Individuals with tetraplegia had significantly higher (p < .05) measures of kyphosis (42 + 16.0 ) and scoliosis (14 degrees +/-9.2 degrees) than the C subjects (kyphosis, 32 degrees +/- 7.9 degrees ; scoliosis, 5 degrees+/-3.8 degrees). No correlation was found between scores on the SF-MPQ and degree of kyphosis or scoliosis. Significant differences were seen between the NT and OT groups on both CES-D (NT, 15.2+/-8.1; OT, 5.8+/ 5.5) and LSIA (NT, 9.9+/-2.8; OT, 14.4+/-2.9). CONCLUSION: This study indicates that seated kyphosis and scoliosis develop early in individuals with tetraplegia and may not be progressive. No association was seen between pain and kyphosis or scoliosis in this relatively young sample (mean age of OT and NT combined, 34.8 years). Future research is needed to determine whether pain becomes a problem in individuals with significant kyphosis or scoliosis as they age. PMID- 9862305 TI - Age-related changes in the initiation of gait: degradation of central mechanisms for momentum generation. AB - OBJECTIVE: To investigate cross-sectionally age-related changes in the expression and biomechanical efficiency of the gait-initiation motor program. DESIGN: Case control study. PARTICIPANTS AND SETTING: Twenty healthy young research subjects and 20 healthy elderly subjects who volunteered from the community participated in this study at a university research laboratory. MAIN OUTCOME MEASURES: Participants performed gait-initiation trials at three speeds from a starting position on a force platform while ground reaction force data, 3-D motion analysis data, and electromyographic data were collected. Measures included: latency of tibialis anterior (TA) activation and soleus (SOL) and gastrocnemius (GA) inhibition, magnitude of center of pressure (COP) displacement, magnitude of momentum generated, and final walking velocity. RESULTS: The expression of the central motor program governing gait initiation, as evidenced by the invariant timing between TA activation and SOL/GA inhibition, was seen in both the young and elderly populations, but the frequency was diminished in the latter group. The momentum-generating capacity of the COP shift mechanism was present but significantly diminished in the elderly population. CONCLUSIONS: These findings suggest that the central nervous system uses stable, efficient mechanisms for dealing with the inherent instability of upright bipedalism and that the integrity of these mechanisms degrades with aging. PMID- 9862306 TI - Osteogenesis imperfecta in childhood: treatment strategies. AB - Osteogenesis imperfecta (OI) is a skeletal disorder of remarkable clinical variability characterized by bone fragility, osteopenia, variable degrees of short stature, and progressive skeletal deformities. Additional clinical manifestations such as blue sclerae, dentinogenesis imperfecta, joint laxity, and maturity onset deafness are described in the literature. OI occurs in about 1 in 20,000 births and is caused by quantitative and qualitative defects in the synthesis of collagen I. Depending on the severity of the disease, a large impact on motor development, range of joint motion, muscle strength, and functional ability may occur. Treatment strategies should primarily focus on the improvement of functional ability and the adoption of compensatory strategies, rather than merely improving range of joint motion and muscle strength. Surgical treatment of the extremities may be indicated to stabilize the long bones to optimize functional ability and walking capacity. Surgical treatment of the spine may be indicated in patients with progressive spinal deformity and in those with symptomatic basilar impression. PMID- 9862307 TI - Dermatomyositis associated with fallopian tube carcinoma. AB - Dermatomyositis (DM) is an idiopathic inflammatory myopathy associated with characteristic cutaneous and extracutaneous manifestations, including malignancy. Primary fallopian tube carcinoma (FTC) is the least common site of origin for a malignant neoplasm of the female genital tract. This report describes the first documented case of DM and concurrent FTC in the United States. A 62-year-old woman presented with DM and was subsequently found to have FTC. During her clinical course, she had improvement in strength and function with treatment of the underlying FTC, which also correlated with lower creatinine phosphokinase levels. An association between DM and FTC may exist because of significant functional strength improvements after tumor removal and chemotherapy. PMID- 9862308 TI - The elixir of laughter in rehabilitation. PMID- 9862309 TI - Endophthalmitis. AB - Endophthalmitis is an inflammatory reaction of intraocular fluids or tissues. Infectious endophthalmitis is one of the most serious complications of ophthalmic surgery. Occasionally, infectious endophthalmitis is the presenting feature of an underlying systemic infection. Successful management of infectious endophthalmitis depends on timely diagnosis and institution of appropriate therapy. Recognition of the different clinical settings in which endophthalmitis occurs and awareness of the highly variable presentation it may have facilitate timely diagnosis. Biopsy of intraocular fluid/tissue is the only method that permits reliable diagnosis and treatment. The different presenting clinical settings, a rational approach to diagnosis (i.e., when, what, and how to biopsy), and the treatment of infectious endophthalmitis are reviewed. PMID- 9862310 TI - Conjunctivochalasis: literature review and possible pathophysiology. AB - Conjunctivochalasis, defined as a redundant conjunctiva typically located between the globe and the lower eyelid, is not uncommon. However, it is often overlooked, as it may be considered a normal senile change. Conjunctivochalasis can cause a spectrum of symptoms, ranging from aggravation of a dry eye at the mild stage, to disturbance of tear outflow at the moderate stage, and exposure problems at the severe stage. Clinical recognition of its pathogenic role helps differentiate conjunctivochalasis from other diseases that may generate similar symptoms. For symptomatic patients, topical lubricants can be tried, but they are frequently unsuccessful, and surgical excision may be required. To guide future investigation into the clinical significance of conjunctivochalasis, a comprehensive grading system and a hypothetical pathophysiology are proposed. Special attention is given to the relationship of conjunctivochalasis to tear dynamics. PMID- 9862311 TI - Serum lipid physiology and the influence of glaucoma medications. AB - Understanding the effect of serum lipid levels on risk factors for coronary heart disease and how they are influenced by medical therapy may lead to overall better care of the glaucoma patient. Elevated low-density lipoprotein and total cholesterol levels are major risk factors for heart disease. In contrast, high density lipoprotein (HDL) is protective for heart disease. beta-adrenergic blockers, a class of medicines used to treat glaucoma, may influence serum lipid levels. Oral nonselective beta-adrenergic blockers reduce HDL cholesterol by 19% and increase triglycerides by 20-40%. Furthermore, topical nonselective beta adrenergic blockers also decrease serum HDL and worsen the total cholesterol/HDL ratio. However, beta-blockers with intrinsic sympathomimetic activity appear to be lipid neutral. At present, there is no clear clinical evidence to indicate that changes in serum lipids with use of topical beta-adrenergic blockers significantly affect the clinical course of the patient. Little information is available for other classes of medicines used topically to treat glaucoma. However, oral preparations of prostaglandins, alpha-adrenergic agonists, angiotensin-converting enzyme inhibitors, and calcium channel blockers do not adversely affect serum lipid levels. Further study is required on newer glaucoma preparations to determine their specific actions on lipid levels. Additionally, further work is required to understand the significance of not only the adverse effect of beta-adrenergic blockers on lipid levels, but their overall effect on long-term cardiac morbidity and mortality. PMID- 9862312 TI - Ocular neovascularization: an epidemiologic review. AB - Neovascularization occurs in many eye diseases, and its epidemiologic impact is significant. However, data on the prevalence and incidence of ocular neovascularization have never been compiled to demonstrate its pervasiveness. This overview of ocular angiogenesis provides a review of the epidemiologic literature for neovascularization in various parts of the eye, including the cornea, iris, retina, and choroid. Relevant disease states are reviewed, as are their risk factors, so that their pathogenesis can be better understood. Data on the prevalence and incidence of the major diseases involving angiogenesis are synthesized to provide statistical evidence of the span and magnitude of ocular neovascularization. These prevalence and incidence data on ocular neovascularization are extrapolated to USA population data where possible, and "worst-case" estimates are calculated as well. Information was gathered with a search of the MEDLINE database, published monographs and volumes, and consultation with a number of primary authors. This study attempts to unify much of past and present epidemiologic research, and the information is presented in sections divided according to the anatomy of the eye. PMID- 9862313 TI - Optic disk edema with a macular star. AB - A 13-year-old boy presented with acute loss of vision in his right eye of 2 weeks' duration. He had a high fever and was ill for several days, then improved but suffered recurrent episodes of sweating and a high fever. Ophthalmoscopy of the right eye showed optic disk edema, mild vitreous cells, and minimal exudates in the macula. Bartonella henselae titers were positive. A diagnosis of optic disk edema with a macular star secondary to cat-scratch disease was made. The patient was treated with doxycycline and made a dramatic improvement to visual acuity of 20/30 with a minimal residual relative central scotoma. The optic disk edema and macular star resolved, and the patient was left with mild optic atrophy in the right eye. PMID- 9862314 TI - The use of dissimilar progressives in the management of presbyopia. AB - The management of anisophoria with bifocal spectacle correction has always presented a challenging refractive problem. Historically, practitioners have used dissimilar bifocal segments or bicentric grinding to neutralize induced vertical imbalance. We present a method of reducing anisophoria at near point with dissimilar progressive multifocals. By incorporating different progressive designs, anisophoria can be minimized to a tolerable level, based on the dioptric power of lens and progressive lens design. PMID- 9862315 TI - Opportunities for maloccurrence in delivery of specialized care in the managed care environment. AB - Two patients lost eyes because of complications of retinal detachment in managed care situations. In both cases the eyes might have been saved had the patients been treated earlier by highly qualified subspecialists. PMID- 9862316 TI - Epidemiology of blindness and eye disease. PMID- 9862317 TI - Cancer incidence after retinoblastoma: radiation dose and sarcoma risk. PMID- 9862318 TI - Recommendations of the Optic Neuritis Study Group. PMID- 9862319 TI - Toward a new neurobiology of energy balance, appetite, and obesity: the anatomists weigh in. PMID- 9862321 TI - Hypocretin/orexin- and melanin-concentrating hormone-expressing cells form distinct populations in the rodent lateral hypothalamus: relationship to the neuropeptide Y and agouti gene-related protein systems. AB - Cells in the lateral hypothalamus and in the arcuate nucleus play prominent roles in the central control of food intake; however, a neurochemical link connecting these potential components of a hypothalamic circuitry regulating energy metabolism remains to be established. In the present study, the topographical relationship between cells expressing mRNAs encoding melanin-concentrating hormone and the newly discovered neuropeptide family hypocretins/orexins was studied in the rat and mouse lateral hypothalamus by using double-labeling in situ hybridization. Cells expressing the two mRNAs formed completely distinct populations, with hypocretin/orexin cells located primarily perifornically and in the magnocellular lateral hypothalamic nucleus; melanin-concentrating hormone cells extended in a wider area both laterally and periventricularly and appeared to partly surround the hypocretin/orexin population. In the arcuate nucleus, cells expressing neuropeptide Y and agouti gene-related protein were studied by routine fluorescence and/or confocal microscopy immunohistochemistry. Double staining demonstrated that a large proportion of the neuropeptide Y-positive cell bodies in this nucleus also contained agouti gene-related protein-like immunoreactivity. Moreover, these two peptides also coexisted in nerve terminals surrounding and in close relationship to perikarya and processes of both hypocretin/orexin- and melanin-concentrating hormone-immunoreactive cells in the lateral hypothalamus, whereby the former appeared to receive a more dense innervation. These results thus provide evidence for an arcuate-lateral hypothalamic neuropeptide Y/agouti gene-related protein pathway. Furthermore, the results implicate hypocretin/orexin and melanin-concentrating hormone-expressing cells as downstream targets in neuropeptide Y-induced feeding. PMID- 9862320 TI - Chemically defined projections linking the mediobasal hypothalamus and the lateral hypothalamic area. AB - Recent studies have identified several neuropeptide systems in the hypothalamus that are critical in the regulation of body weight. The lateral hypothalamic area (LHA) has long been considered essential in regulating food intake and body weight. Two neuropeptides, melanin-concentrating hormone (MCH) and the orexins (ORX), are localized in the LHA and provide diffuse innervation of the neuraxis, including monosynaptic projections to the cerebral cortex and autonomic preganglionic neurons. Therefore, MCH and ORX neurons may regulate both cognitive and autonomic aspects of food intake and body weight regulation. The arcuate nucleus also is critical in the regulation of body weight, because it contains neurons that express leptin receptors, neuropeptide Y (NPY), alpha-melanin stimulating hormone (alpha-MSH), and agouti-related peptide (AgRP). In this study, we examined the relationships of these peptidergic systems by using dual label immunohistochemistry or in situ hybridization in rat, mouse, and human brains. In the normal rat, mouse, and human brain, ORX and MCH neurons make up segregated populations. In addition, we found that AgRP- and NPY-immunoreactive neurons are present in the medial division of the human arcuate nucleus, whereas alpha-MSH-immunoreactive neurons are found in the lateral arcuate nucleus. In humans, AgRP projections were widespread in the hypothalamus, but they were especially dense in the paraventricular nucleus and the perifornical area. Moreover, in both rat and human, MCH and ORX neurons receive innervation from NPY , AgRP-, and alpha-MSH-immunoreactive fibers. Projections from populations of leptin-responsive neurons in the mediobasal hypothalamus to MCH and ORX cells in the LHA may link peripheral metabolic cues with the cortical mantle and may play a critical role in the regulation of feeding behavior and body weight. PMID- 9862322 TI - Widespread expression in adult rat forebrain of mRNA encoding high-affinity neurotensin receptor. AB - The peptides neurotensin (NT) and neuromedin N exert effects on neurons by means of a high-affinity NT receptor (NTRH) belonging to the superfamily of G-protein coupled receptors. In the present study, we used in situ hybridization histochemistry with sensitive riboprobe methodology to investigate the distribution of NTRH mRNA in the forebrain of adult rats. Labeled cells were abundant in the hypothalamus, epithalamus, ventral thalamus, septum, amygdala, and pallidum, including many regions where NTRH mRNA had not been detected previously. In the hypothalamus, novel sites of NTRH mRNA expression included the arcuate, periventricular, paraventricular, supraoptic, medial preoptic, anterior, ventromedial, and posterior nuclei, as well as the lateral hypothalamic area. In the thalamus, novel sites of expression included the anterodorsal nucleus, lateral habenula, and zona incerta, where labeling was much more extensive than previously reported. Novel telencephalic sites of expression included most bed nuclei of the stria terminalis, most divisions of the amygdala, the main olfactory bulb, the endopiriform nucleus, the claustrum, many parts of retrohippocampal allocortex, and limited parts of most isocortical areas. Novel sites of expression were also observed in the midbrain and pons. Taking into account expected differences in the subcellular locations of receptor mRNA and protein, the regional distribution of NTRH mRNA agrees well with that of NTRH determined previously. Our results identify many novel sites of NTRH mRNA expression in adult brain and provide a basis for investigating involvement of NT and related peptides in regulating the activity of these diverse cells, whose phenotypes remain largely undetermined. PMID- 9862323 TI - Mushroom bodies of the cockroach: activity and identities of neurons recorded in freely moving animals. AB - This article describes novel attributes of the mushroom bodies of cockroaches revealed by recording from neurons in freely moving insects. The results suggest several hitherto unrecognized functions of the mushroom bodies: extrinsic neurons that discriminate between imposed and self-generated sensory stimulation, extrinsic neurons that monitor motor actions, and a third class of extrinsic neurons that predict episodes of locomotion and modulate their activity depending on the turning direction. Electrophysiological units have been correlated with neurons that were partially stained by uptake of copper ions and silver intensification. Neurons so revealed correspond to Golgi-impregnated or Lucifer yellow-filled neurons and demonstrate that their processes generally ascend to other areas of the protocerebrum. The present results support the idea of multiple roles for the mushroom bodies. These include sensory discrimination, the integration of sensory perception with motor actions, and, as described in the companion article, a role in place memory. PMID- 9862325 TI - Interhemispheric connections of somatosensory cortex in the flying fox. AB - The interhemispheric connections of somatosensory cortex in the gray-headed flying fox (Pteropus poliocephalus) were examined. Injections of anatomical tracers were placed into five electrophysiologically identified somatosensory areas: the primary somatosensory area (SI or area 3b), the anterior parietal areas 3a and 1/2, and the lateral somatosensory areas SII (the secondary somatosensory area) and PV (pairetal ventral area). In two animals, the hemisphere opposite to that containing the injection sites was explored electrophysiologically to allow the details of the topography of interconnections to be assessed. Examination of the areal distribution of labeled cell bodies and/or axon terminals in cortex sectioned tangential to the pial surface revealed several consistent findings. First, the density of connections varied as a function of the body part representation injected. For example, the area 3b representation of the trunk and structures of the face are more densely interconnected than the representation of distal body parts (e.g., digit 1, D1). Second, callosal connections appear to be both matched and mismatched to the body part representations injected in the opposite hemisphere. For example, an injection of retrograde tracer into the trunk representation of area 3b revealed connections from the trunk representation in the opposite hemisphere, as well as from shoulder and forelimb/wing representations. Third, the same body part is differentially connected in different fields via the corpus callosum. For example, the D1 representation in area 3b in one hemisphere had no connections with the area 3b D1 representation in the opposite hemisphere, whereas the D1 representation in area 1/2 had relatively dense reciprocal connections with area 1/2 in the opposite hemisphere. Finally, there are callosal projections to fields other than the homotopic, contralateral field. For example, the D1 representation in area 1/2 projects to contralateral area 1/2, and also to area 3b and SII. PMID- 9862324 TI - Mushroom bodies of the cockroach: their participation in place memory. AB - Insects and other arthropods use visual landmarks to remember the location of their nest, or its equivalent. However, so far, only olfactory learning and memory have been claimed to be mediated by any particular brain region, notably the mushroom bodies. Here we describe the results of experiments that demonstrate that the mushroom bodies of the cockroach (Periplaneta americana), already shown to be involved in multimodal sensory processing, play a crucial role in place memory. Behavioral tests, based on paradigms similar to those originally used to demonstrate place memory in rats, demonstrate a rapid improvement in the ability of individual cockroaches to locate a hidden target when its position is provided by distant visual cues. Bilateral lesions of selected areas of the mushroom bodies abolish this ability but leave unimpaired the ability to locate a visible target. The present results demonstrate that the integrity of the pedunculus and medial lobe of a single mushroom body is required for place memory. The results are comparable to the results obtained from hippocampal lesions in rats and are relevant to recent studies on the effects of ablations of Drosophila mushroom bodies on locomotion. PMID- 9862326 TI - Lack of Th2 cytokine increase during spontaneous remission of experimental allergic encephalomyelitis. AB - The mechanisms underlying spontaneous remission of autoimmune diseases are presently unknown, though regulatory T cells are believed to play a major role in this process. We tested the hypothesis that Th2 and/or other T cell regulatory cytokines cause the spontaneous remission of experimental allergic encephalomyelitis (EAE), a model of Th1-mediated autoimmunity. We analyzed the cytokine profile of lymph node and central nervous system-infiltrating cells in individual SJL mice at different stages of proteolipid protein (PLP) 139-151 peptide-induced EAE. We found that IFN-gamma slowly fades away after clinical recovery, whereas IL-4, IL-10 and transforming growth factor-beta remain low or undetectable. Our peptide-results therefore suggest that regulatory T cells producing anti-inflammatory cytokines are not involved in spontaneous remission of EAE and challenge the view that the Th1/Th2 balance has a key role in EAE regulation. PMID- 9862327 TI - Local gene therapy with CTLA4-immunoglobulin fusion protein in experimental allergic encephalomyelitis. AB - It has been reported previously that the induction phase of experimental allergic encephalomyelitis (EAE) is highly sensitive to systemic blockade of stimulation via MHC class II molecules and co-stimulation via the CD28:CD80/CD86 pathways. In contrast, the effector phases of EAE were relatively unaffected by similar treatments using MHC class II antigen (Ag)-specific mAb and cytotoxic T lymphocyte antigen (CTLA)4-Ig fusion proteins in some studies. This has been attributed to different sensitivities of effector cell function or the poor penetrance of inhibitory proteins into the central nervous system (CNS). To examine this question further, MHC class II Ag-specific mAb and CTLA4-Ig were delivered directly into the CNS following EAE induction, and both were found to inhibit disease. While it was found that systemic administration of mouse CTLA4 Ig could also inhibit the progression of effector immune responses when administered shortly before or during clinical disease, these were significantly more active when delivered directly into the CNS, which probably involved an action on both CD28 ligands, CD80 and CD86. Although mouse CTLA4-human Ig was therapeutically less efficient than mouse CTLA4-mouse Ig protein, probably due to the enhanced immunogenicity and lower functional activity, gene delivery of CTLA4 human Ig into the CNS using a non-replicating adenoviral vector was more effective than a single injection of CTLA4-human Ig protein. Gene delivery significantly ameliorated the development of EAE, without necessarily inhibiting unrelated peripheral immune responsiveness. Local gene delivery of CTLA4-Ig may thus be an important target for immunotherapy of human autoimmune conditions such as multiple sclerosis. PMID- 9862328 TI - Human preformed IgG combining with membrane-bound porcine serotransferrin lyse porcine endothelial cells through antibody-dependent cellular cytotoxicity. AB - Preformed antibodies are involved in xenograft rejection. The purpose of this work was to characterize porcine xenoantigens recognized by human preformed IgG (hpIgG), and to investigate the role of hpIgG in xenogeneic rejection. IgG eluted from porcine livers perfused with human plasma, human sera and total human IgG were immunoblotted on porcine aortic endothelial cell extracts. The amino acid sequence of a 76-kDa antigen constantly revealed was 100% homologous with porcine serotransferrin (psTf). hpIgG from human sera, human IgG1 and IgG2 and F(ab')2gamma specifically bound to psTf. Neutralization by psTf abolished that binding. Although alpha1,3-linked galactose residues (Gal(alpha)1,3Gal) is the dominant epitope recognized by preformed antibodies in the swine-to-human combination, the analysis of carbohydrate composition of psTf showed that the molecule was devoid of Gal(alpha)1,3Gal moieties and that preformed anti-psTf IgG bound to epitopes localized on the peptide core of the molecule. Purified human anti-psTf IgG antibodies were able to bind to psTf linked to its receptor on porcine endothelial cells, and to kill those cells through antibody-dependent cellular cytotoxicity. PMID- 9862329 TI - Modulation of calcium responses by altered peptide ligands in a human T cell clone. AB - To determine whether altered peptide ligands (APL) affect calcium signaling events, we investigated changes in intracellular calcium concentration ([Ca2+]i) in human T cell clone stimulated with either the fully agonistic peptide M12p54 68, the partially agonistic analogue E63V or the simple antagonistic analogue E58M. Both E63V and E58M stimulated a Ca2+ response in approximately 40% of T cells, whereas M12p54-68 did so in approximately 70% of T cells. The most predominant pattern of a Ca2+ increase induced by M12p54-68 was a small sinusoidal peak followed by a sustained high response. The most frequent pattern of calcium response induced by E63V was a continuous high response without a preceding sinusoidal peak, whereas that induced by E58M was large with frequent oscillations. Genistein, an inhibitor of the protein tyrosine kinases (PTK), markedly inhibited the wild-type peptide-induced increase in [Ca2+]i, whereas it marginally inhibited the response induced by E63V or E58M. In contrast, GF109203X, a protein kinase C (PKC)-specific inhibitor, markedly inhibited the E63V- or E58M-induced Ca2+ response, whereas it marginally affected the wild peptide-induced Ca2+ response. Furthermore, in nominal Ca2+-free medium, the E58M induced Ca2+ response was almost completely blocked, while the M12p54-68- or E63V induced responses were only partially inhibited. Our results suggest that the Ca2+ response induced by the fully agonistic peptide depends on activation of the genistein-sensitive signaling pathway, including PTK, whereas the Ca2+ response to a simple antagonistic APL completely depends on extracellular Ca2+ and activation of the GF109203X-sensitive signaling pathway, including PKC. These differences in the CA2+i response in recognition of different APL may parallel the unique T cell activation patterns induced by APL in human T cells. PMID- 9862330 TI - Enhancement of terminal B lymphocyte differentiation in vitro by fibroblast-like stromal cells from human spleen. AB - Stromal elements are major components of lymphoid tissues contributing to both tissue architecture and function. In this study we report on the phenotype and function of fibroblast-like stromal cells obtained from human spleen. These cells express high levels of CD44 and ICAM-1 and moderate levels of VLA-4, VCAM, CD40 and CD21. They fail to express endothelial, epithelial, lymphocyte and monocyte/macrophage markers. We show that these cells interact with B cell blasts induced in vitro by anti-CD40 and anti-mu stimulation. As a result of these interactions both IL-6 and IgG secretion into culture medium is increased. The enhanced secretion of IgG is partly inhibited by abolishing B cell blaststromal cell contact or by anti-IL-6, anti-VCAM or anti-CD49d antibodies. Our studies also suggest that the ability of stromal cells to promote B cell survival is most likely the underlying mechanism of the enhanced immunoglobulin secretion. Comparison of stromal cells from different lymphoid and non-lymphoid organs revealed that bone marrow- and spleen-derived stromal cells are the most effective in promoting B cell blast differentiation. PMID- 9862331 TI - Differential T cell responses to Mycobacterium tuberculosis ESAT6 in tuberculosis patients and healthy donors. AB - Vaccination against and diagnosis of tuberculosis are still insufficient. Proteins secreted by Mycobacterium tuberculosis induce strong immune responses in tuberculosis and constitute prime candidates for development of novel vaccines against tuberculosis as well as for immunodiagnostic assays. We investigated the role of the secreted proteins MPT63, MPT64 and ESAT6 from M. tuberculosis in healthy individuals and tuberculosis patients. None of the secreted proteins stimulated peripheral blood mononuclear cells from healthy donors. In contrast, CD4+ T cells from many tuberculosis patients were stimulated in an MHC class II restricted fashion by ESAT6, but not by MPT63 or MPT64. T cell reactivities of tuberculosis patients were focused on the N-terminal region of ESAT6. The ESAT6 T cell epitopes were presented by different HLA-DR phenotypes. Cell cultures responding to either ESAT6 or synthetic peptides thereof showed mRNA transcripts for macrophage inflammatory protein (MIP)-1alpha, monocyte chemotactic protein (MCP)-1 or IL-8 and production of IFN-gamma and MIP-1alpha. Our results suggest that the secreted M. tuberculosis proteins MPT63, MPT64 or ESAT6 do not stimulate unprimed T cells, and that ESAT6 may be a potential candidate antigen for detection of clinical disease. PMID- 9862332 TI - Isotypic variation of novel immunoglobulin-like transcript/killer cell inhibitory receptor loci in the leukocyte receptor complex. AB - The leukocyte receptor complex (LRC) on human chromosome 19q13.4 encompasses at least four families of related genes: immunoglobulin-like transcripts (ILT), killer cell inhibitory receptors (KIR), the leukocyte-associated inhibitory receptors (LAIR) and the Fcalpha receptor (Fc(alpha)R). We determined the genomic organization of a region of DNA spanning the junction of the ILT and KIR gene complexes. Extensive sequence data were collected for ILT3, two novel genes, ILT9 and ILT10, and one novel KIR locus (KIRCI). These loci, along with other reported sequences from the region, encoded a leader sequence split into two exons, upstream of two to four immunoglobulin (Ig) domains, each on a separate exon. Downstream of the Ig domains, however, the organization differs markedly between inhibitory and activating ILT. These data are consistent with a highly conserved gene arrangement for all superfamily members suggesting duplication of primordial sequences. ILT3 and KIRCI were in the same head-to-tail orientation as has been described for other KIR loci which may facilitate addition or loss of genes between different haplotypes. PMID- 9862333 TI - Double-stranded deoxyribonucleic acid binds to HLA class II molecules and inhibits HLA class II-mediated antigen presentation. AB - CD4+ T cells proliferating in response to purified double-stranded deoxyribonucleic acid (dsDNA) have been recently demonstrated in peripheral blood mononuclear cells of patients with systemic lupus erythematosus. Their activation was inhibited by anti-HLA class II (HLA-II) monoclonal antibodies; thus, the existence of a molecular interaction between dsDNA and HLA-II is conceivable. In this report we show that dsDNA specifically bind to HLA-II. After preincubating cells with purified dsDNA or synthetic oligonucleotides, dsDNA was detected on the cell membrane and in the lysates of HLA-II+ but not of isogenic HLA-II- cell lines. We demonstrate that dsDNA binding inhibits that of a specific peptide to HLA-II. Mixed lymphocyte reaction and antigen-specific T cell proliferation were inhibited by the preincubation of stimulator cells or antigen-presenting cells with dsDNA. These results suggest the existence of a novel mechanism of down modulation of the CD4+ T cell function generated by lack of stimulation due to the HLA-II presenting molecules being "occupied" by dsDNA. PMID- 9862334 TI - Regulation of IFN-gamma production in granulocyte-macrophage colony-stimulating factor-deficient mice. AB - Granulocyte-macrophage colony-stimulating factor-deficient (GM-CSF-/-) mice produce far lower serum levels of IFN-gamma in response to LPS than GM-CSF+/+ mice. CD4+ and CD8+ T cells from LPS-injected GM-CSF-/- mice showed a deficiency in IFN-gamma production and proliferative activity in response to IL-2 and IL-12, whereas IFN-gamma production by NK cells was not compromised. These defects of T cells were reversed by administration of GM-CSF in vivo, but not by supplementation with GM-CSF in vitro. GM-CSF-/- mice do not have an intrinsic defect in IFN-gamma production, because IL-12 injection induces the same high levels of IFN-gamma in GM-CSF-/- and GM-CSF+/+ mice. To investigate the inhibitory effect of LPS on GM-CSF-/- T cells and the indirect restorative activity of GM-CSF, we tested the action of supernatants from cultured dendritic cells (DC). A factor or factors in the DC supernatant normalized serum IFN-gamma levels and T cell responses in LPS-injected GM-CSF-/- mice. IL-18 reproduced some but not all of these in vivo and in vitro effects of DC supernatants. Our results indicate that GM-CSF is important in protecting T cells from inhibitory signals generated during immunization or exposure to LPS, and that this effect of GM-CSF is indirect and mediated by factors produced by DC. PMID- 9862335 TI - Differential reactivity of brain microvascular endothelial cells to TNF reflects the genetic susceptibility to cerebral malaria. AB - Upon infection with Plasmodium berghei ANKA (PbA), various inbred strains of mice exhibit different susceptibility to the development of cerebral malaria (CM). Tumor necrosis factor-alpha (TNF) and interferon-gamma (IFN-gamma) have been shown to be crucial mediators in the pathogenesis of this neurovascular complication. Brain microvascular endothelial cells (MVEC) represent an important target of both cytokines. In the present study, we show that brain MVEC purified from CM-susceptible (CM-S) CBA/J mice and CM-resistant (CM-R) BALB/c mice exhibit a different sensitivity to TNF. CBA/J brain MVEC displayed a higher capacity to produce IL-6 and to up-regulate intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in response to TNF than BALB/c brain MVEC. In contrast, no difference was found in the induction of E-selectin after TNF challenge. CM-S brain MVEC were also significantly more sensitive to TNF induced lysis. This differential reactivity to TNF was further substantiated by comparing TNF receptor expression on CM-S and CM-R brain MVEC. Although the constitutive expression of TNF receptors was comparable on cells from the two origins, TNF induced an up-regulation of both p55 and p75 TNF receptors in CM-S, but not in CM-R brain MVEC. A similar regulation was found at the level of TNF receptor mRNA, but not for receptor shedding. Although a protein kinase C inhibitor blocked the response to TNF in both the brain MVEC, an inhibitor of protein kinase A selectively abolished the response to TNF in CM-R, but not CM-S brain MVEC, suggesting a differential protein kinase involvement in TNF-induced activation of CM-S and CM-R brain MVEC. These results indicate that brain MVEC purified from CM-S and CM-R mice exhibit distinctive sensitivity to TNF This difference may be partly due to a differential regulation of TNF receptors and via distinct protein kinase pathways. PMID- 9862336 TI - Human amniotic IgA inhibits natural IgG autoantibodies of maternal or unrelated origin. AB - We show that the natural autoantibody activity of amniotic IgG dramatically increases after purification, and that the IgG-depleted fraction can suppress the activity of IgG natural antibodies from amniotic fluid or from the maternal serum. This suppression is also observed towards serum IgG from unrelated adults but does not impair the tetanus antitoxin activity of serum-derived IgG. Absorption experiments and immunoglobulin separation by gel permeation demonstrate that this suppression is due to monomeric immunoglobulins of the IgA isotype. The inhibition is associated with an anti-F(ab')2 activity of the amniotic IgA, involving hypervariable regions of the IgG as demonstrated by different reactivities towards monoclonal IgG sharing the same family of VH and Vkappa domains. These results indicate that the inhibition of natural autoantibodies not only occurs with fetal and adult serum IgM, as reported by other groups, but also with amniotic IgA, suggesting a general and important phenomenon. In the case of the amniotic fluid, IgA could protect the fetus against maternal IgG autoantibodies without interfering with simultaneously translocated antigen-induced IgG antibodies to pathogens. PMID- 9862337 TI - The shared tumor-specific antigen encoded by mouse gene P1A is a target not only for cytolytic T lymphocytes but also for tumor rejection. AB - A number of human tumor antigens have been characterized recently using cytolytic T lymphocytes (CTL) as screening tools. Some of them are encoded by MAGE-type genes, which are silent in normal tissues except in male germ cells, but are activated in a variety of tumors. These tumor-specific shared antigens appear to be promising targets for cancer immunotherapy. However, the choice of these antigens as targets has been questioned because of the lack of direct evidence that in vivo responses against such antigens can lead to tumor rejection. The antigen encoded by the mouse gene P1A represents the only available animal model system for MAGE-type tumor antigens. We show here that mice immunized by injection of L1210 leukemia cells expressing P1A and B7-1 (L1210.P1A.B7-1) are efficiently protected against a challenge with a lethal dose of mastocytoma P815 tumor cells, which express P1A. Mice immunized with L1210 cells expressing B7-1 but not P1A were not protected. Furthermore, we observed that P1A-transgenic mice, which are tolerant to P1A, were not protected after immunization with L1210.P1A.B7-1. These results demonstrate that the immune response to P1A is the major component of the tumor rejection response observed in normal mice, and support the use of tumor-specific shared antigens as targets for the immunotherapy of human cancer. PMID- 9862338 TI - Parasite dose determines the Th1/Th2 nature of the response to Leishmania major independently of infection route and strain of host or parasite. AB - Leishmania major causes cutaneous leishmaniasis in mice and man. Infection of mice with relatively low or high numbers of parasites leads respectively to parasite containment, associated with a Th1, cell-mediated response, or progressive disease, associated with a Th2, antibody response in all circumstances studied. These include different parasite strains, different routes of infection, and different hosts previously classified as susceptible, resistant or of intermediate susceptibility. This dose dependency appears to reflect a general rule. We argue that this rule may allow the design of a vaccination strategy that is effective among a genetically diverse population, and that it imposes severe constraints upon proposals for the nature of the "decision criterion" determining whether antigen induces a Th1 or Th2 response. PMID- 9862339 TI - Generation of the vesicular stomatitis virus nucleoprotein cytotoxic T lymphocyte epitope requires proteasome-dependent and -independent proteolytic activities. AB - The proteasome is involved in the generation of most of the MHC class I antigenic epitopes. However, it is not known if the proteasome generates the exact cytotoxic T lymphocyte (CTL) epitope or only epitope precursors which require further modification by additional proteases. Digestion of the extended vesicular stomatitis virus nucleoprotein epitope 52-59 (RGYVYQGL) by the 20S proteasome in vitro shows that the proteasome is capable of generating the correct C terminus but not the exact N terminus of the CTL epitope. This finding suggests that proteolytic activity in addition to the proteasome is required for generation of the CTL epitope. By using the proteasome inhibitor lactacystin we were able to confirm this finding in vivo. Lactacystin prevented the processing of N- and C terminally extended epitopes, whereas the processing of only N-terminally extended epitopes was unaffected. Thus, the proteasome is necessary and sufficient for the generation of the exact C terminus of this CTL epitope, whereas the exact N terminus seems to be generated by a different protease. PMID- 9862340 TI - Influence of B cell receptor ligation and TCR affinity on T-B collaboration in vitro. AB - Co-culture of purified T and B cells obtained from cytochrome c-specific TCR- and hen egg lysozyme (HEL)-specific Ig-transgenic mice was used to examine the role of B cell receptor (BCR) ligation and TCR affinity on the efficiency of T-B cell collaboration. The results showed that BCR ligation of naive B cells with HEL was not required for effective presentation of high-affinity antigen to T cells, although it did enhance activation and division of both T and B cells. Anergic B cells were also effective at presentation of high-affinity antigen and proliferated more than naive B cells in response to T cell help, due to prior exposure to antigen in vivo. Despite the fact that induction of CD86 on anergic B cells following BCR ligation was suboptimal, these cells supported T cell activation and survival in culture as efficiently as naive B cells exposed to HEL. In contrast, when the low-affinity antigen mls-3a served as the T cell stimulus, BCR ligation was essential to elicit a detectable T cell response. Thus the in vitro model demonstrates that co-stimulation is not an absolute requirement for effective antigen presentation and delivery of T cell help to B cells. Rather, the cooperative effects of BCR ligation and TCR affinity determine the relative requirement for co-stimulation. PMID- 9862341 TI - An endogenously processed self peptide and the corresponding exogenous peptide bound to the same MHC class II molecule could be distinct ligands for TCR with different kinetic stability. AB - Immunization with self peptides often elicits activation of CD4+ T cells in vivo. Although such peptides have been suggested to be derived from minor self determinants or self antigens sequestered from the immune system, we found that immunization with Ealpha peptide (Ealpha52-68), a major self determinant bound to I-Ab molecules, elicits an immune response in Ealpha-transgenic C57BL/6 (Ealpha B6) mice where Ealpha52-68 is endogenously processed and presented by I-Ab molecules in the thymus and periphery. To better understand this response, a panel of T cell hybridomas raised against exogenous Ealpha52-68 were analyzed for their reactivity to spleen cells from Ealpha-B6 mice. Some hybridomas were stimulated with Ealpha-B6 spleen cells in the absence of exogenous Ealpha52-68, whereas others were not stimulated with them. The Ealpha52-68/I-Ab complex recognized by the TCR that is expressed on the hybridoma with reactivity to Ealpha-B6 spleen cells was found to be quite stable, whereas the complex recognized by the TCR on the hybridoma specific for the exogenous Ealpha52-68 lost the stimulation activity by incubation the complex at 37 degrees C for 10 min. Stimulation experiments using extensively substituted Ealpha analogue peptides suggested that amino acid residues at positions 57, 58, 60 and 62 of Ealpha52-68 are involved in the interaction with TCR recognizing the Ealpha52 68/I-Ab complex expressed on Ealpha-B6 spleen cells. While amino acid substitutions at positions 60 and 62 also affected the recognition of TCR specific for exogenous Ealpha52-68, all or many amino acid substitutions were allowed at position 58 or 57, respectively, without impairing the TCR recognition. Taken together, these results suggest that endogenously processed self peptide and the corresponding exogenous peptide bound to the same MHC class II molecule could be distinct TCR ligands with different kinetic stability and probably with different configuration. PMID- 9862342 TI - Nitric oxide regulates Th1 cell development through the inhibition of IL-12 synthesis by macrophages. AB - We have previously reported that mice lacking inducible nitric oxide synthase (NOS2) developed enhanced Th1 cell responses. We now investigated the mechanism by which NO modulates Th1 cells differentiation. Peritoneal macrophages from NOS2 deficient mice infected with Leishmania major in vivo or stimulated with IFN gamma or lipopolysaccharide (LPS) in vitro produced significantly higher levels of IL-12 than those from heterozygous or wild-type mice. A macrophage cell line, J774, produced significant amounts of IL-12 following activation with LPS, or LPS plus IFN-gamma. This could be markedly enhanced by the NOS inhibitor L-NG monomethyl arginine (L-NMMA), but profoundly inhibited by the NO-generating compound S-nitroso-N-acetyl-penicillamine (SNAP). The effect of NO in this system is selective, since SNAP enhanced and L-NMMA decreased TNF-alpha synthesis by LPS activated J774 cells. The differential effect of NO on IL-12 and TNF-alpha is at the transcriptional level and is activation dependent. Since IL-12 is a major inducer of Th1 cells which produce IFN-gamma that can activate macrophages to produce IL-12, our data demonstrate that NO can be an inhibitor of this feedback loop, preventing the excessive amplification of Th1 cells which are implicated in a range of immunopathologies. PMID- 9862343 TI - The gp200-MR6 molecule which is functionally associated with the IL-4 receptor modulates B cell phenotype and is a novel member of the human macrophage mannose receptor family. AB - The human gp200-MR6 molecule has previously been shown to have either an antagonistic or agonistic effect on IL-4 function, demonstrated by inhibition of IL-4-induced proliferation of T cells or mimicking of IL-4-induced maturation of epithelium, respectively. We now show that gp200-MR6 ligation can also mimic IL-4 and have an anti-proliferative pro-maturational influence within the immune system, causing up-regulation of co-stimulatory molecules on B lymphocytes. Biochemical analysis and cDNA cloning reveal that gp200-MR6 belongs to the human macrophage mannose receptor family of multidomain molecules. It comprises 1722 amino acids in toto (mature protein, 1695 amino acids; signal sequence, 27 amino acids) organized into 12 external domains (an N-terminal cysteine-rich domain, a fibronectin type II domain and 10 C-type carbohydrate recognition domains), a transmembrane region and a small cytoplasmic C terminus (31 amino acids) containing a single tyrosine residue (Y1679), but no obvious kinase domain. Strong amino acid sequence identity (77%) suggests that gp200-MR6 is the human homologue of the murine DEC-205, indicating that this molecule has much wider functional activity than its classical endocytic role. We also show that the gp200-MR6 molecule is closely associated with tyrosine kinase activity; the link between gp200-MR6 and the IL-4 receptor may therefore be via intracellular signaling pathways, with multifunctionality residing in its extracellular multidomain structure. PMID- 9862344 TI - A novel dendritic cell population in human blood: one-step immunomagnetic isolation by a specific mAb (M-DC8) and in vitro priming of cytotoxic T lymphocytes. AB - Originating from a common progenitor cell, dendritic cells (DC) appear to develop along early branched pathways into various yet ill-defined subpopulations residing at various sites throughout the body where they capture and present antigen in the most professional fashion. Here we give evidence for a unique subpopulation of human DC circulating in blood that account for 0.5-1% of blood leukocytes only, their most specific characteristic being the expression of a cell surface protein recognized by a novel monoclonal antibody (M-DC8) which enables their isolation by a one-step immunomagnetic procedure. The isolated cells (> 97% pure) present morphologically as typical dendritic cells. They express the Fc(gamma)RIII (CD16), so far not found on DC, and avidly phagocytose latex beads as well as opsonized erythrocytes. These cells not only present antigens efficiently to naive T cells but also induce purified CD8+ T cells to become alloantigen-specific cytotoxic cells. Furthermore, when loaded with a tyrosinase-derived peptide they stimulate T cells from normal donors and melanoma patients to exhibit MHC-restricted specific cytotoxicity against melanoma cells. PMID- 9862345 TI - CTX, a Xenopus thymocyte receptor, defines a molecular family conserved throughout vertebrates. AB - CTX, a cortical thymocyte marker in Xenopus, is an immunoglobulin superfamily (Igsf) member comprising one variable and one constant C2-type Igsf domain, a transmembrane segment and a cytoplasmic tail. Although resembling that of the TCR and immunoglobulins, the variable domain is not encoded by somatic rearrangement of the gene but by splicing of two half-domain exons. The C2 domain, also encoded by two exons, has an extra pair of cysteines. The transmembrane segment is free of charged residues, and the cytoplasmic tail (70 amino acids) contains one tyrosine and many glutamic acid residues. ChT1, a chicken homologue of CTX, has the same structural and genetic features, and both molecules are expressed on the thymocyte surface. We cloned new mouse (CTM) and human (CTH) cDNA and genes which are highly homologous to CTX/ChT1 but not lymphocyte specific. Similarity with recently described human cell surface molecules, A33 antigen and CAR (coxsackie and adenovirus 5 receptor), and a number of expressed sequence tags leads us to propose that CTX defines a novel subset of the Igsf, conserved throughout vertebrates and extending beyond the immune system. Strong homologies within vertebrate sequences suggest that the V and C2 CTX domains are scions of a very ancient lineage. PMID- 9862346 TI - Involvement of Fas/Fas ligand system-mediated apoptosis in the development of concanavalin A-induced hepatitis. AB - Concanavalin A (Con A)-induced hepatitis is an experimental hepatitis model in which hepatic injury is caused by the action of cytokines produced by T cells. Using IFN-gamma-deficient mice, we previously demonstrated that IFN-gamma plays a central role in Con A-induced hepatitis. Here, we show that development of the disease is completely suppressed in gld/gld mice, in which Fas ligand is defective. In contrast, suppression of the disease in Ipr/Ipr mice was incomplete, since a small amount of the fas mRNA was produced in these mice. The data indicate that activation of the Fas/Fas ligand system is a necessary step in the development of Con A-induced hepatitis. Furthermore, we found that not only fas but also caspase-1 expression was reduced in IFN-gamma-deficient mice. Since caspase-1 is an integral component of Fas signal transduction, these observations suggest that IFN-gamma-induced activation of both fas and caspase-1 expression causes enhancement of hepatocyte apoptosis resulting in the development of hepatitis. PMID- 9862347 TI - Dendritic cell chemotaxis and transendothelial migration are induced by distinct chemokines and are regulated on maturation. AB - The capacity of dendritic cells (DC) to initiate immune responses is dependent on their specialized migratory and tissue homing properties. Chemotaxis and transendothelial migration (TEM) of DC were studied in vitro. Immature DC were generated by culture of human monocytes in granulocyte-macrophage colony stimulating factor and IL-4. These cells exhibited potent chemotaxis and TEM responses to the CC chemokines macrophage inflammatory protein (MIP)-1alpha, MIP 1beta, RANTES, and monocyte chemotactic protein-3, and weak responses to the CC chemokine MIP-3beta and the CXC chemokine stromal cell-derived factor (SDF) 1alpha. Maturation of DC induced by culture in lipopolysaccharide, TNF-alpha or IL-1beta reduced or abolished responses to the former CC chemokines but markedly enhanced responses to MIP-3beta and SDF-1alpha. This correlated with changes in chemokine receptor expression: CCR5 expression was reduced while CXCR4 expression was enhanced. These findings suggest two stages for regulation of DC migration in which one set of chemokines may regulate recruitment into or within tissues, and another egress from the tissues. PMID- 9862348 TI - Gamma-glutamyl transpeptidase, an ecto-enzyme regulator of intracellular redox potential, is a component of TM4 signal transduction complexes. AB - CD21 (C3dg/EBV receptor) is physically associated on B cells with a complex of proteins that includes CD19 and the widely distributed tetraspan 4 (TM4) family protein CD81 as well as other TM4 proteins (CD53, CD37 and CD82). Monoclonal antibodies (mAb) were generated that blocked homotypic adhesion induced by CD21 ligands in the human B cell line Balm-1. One inhibitory mAb (3A8) was found to recognize the ecto-enzyme gamma-glutamyl transpeptidase (GGT), a membrane protein involved in recycling extracellular glutathione and regulating intracellular redox potential. Molecular associations between GGT and TM4 proteins CD81, CD53 and CD82, in addition to CD21 and CD19, were detected by co-precipitation and co capping analysis. GGT is expressed on several B and T cell lines independently of CD21 expression. These results demonstrate that GGT is a component of widely distributed TM4 complexes, and that on B cells the GGT-containing TM4 complexes also contain CD19 and CD21. PMID- 9862349 TI - The combined inactivation of tumor necrosis factor and interleukin-6 prevents induction of the major acute phase proteins by endotoxin. AB - The constellation of changes known as the acute phase response (APR) is a cytokine-driven process initiated by tissue inflammation. The proinflammatory cytokines, TNF, IL-1 and IL-6, are considered to be the primary mediators of the APR. IL-6 and IL-1beta gene-deleted mice (Fattori et al., J. Exp. Med. 1994. 180: 1243-1250; Kopf et al., Nature 1994. 368: 339-342; Fantuzzi et al., J. Immunol. 1996. 157: 291-296, respectively), exhibit impaired APR to turpentine injection but only a slight reduction in plasma acute phase protein levels in response to lipopolysaccharide (LPS). This infers an important role for TNF in the LPS induced APR, however, in the present study, normal APR to both turpentine and LPS were observed in TNF/LTalpha-deficient mice. A striking absence of elevated major acute phase proteins, SAP and SAA, was observed in mice deficient in TNF/LTalpha and IL-6, suggesting that TNF-alpha or LTalpha do indeed exert important nonredundant synergism in the IL-1/IL-6 primary response. The regulation of other parameters typically altered in an APR, body weight, blood glucose and haptoglobin, was normal in LPS-dosed TNF/LTalpha-deficient and wild-type mice. The observed transcriptional response for SAA and SAP in these TNF/LTalpha/IL-6 deficient mice, in lieu of elevated plasma levels, suggests that SAA and SAP expression is possibly posttranscriptionally regulated. PMID- 9862350 TI - Influence of maternal antibodies on vaccine responses: inhibition of antibody but not T cell responses allows successful early prime-boost strategies in mice. AB - The transfer of maternal antibodies to the offspring and their inhibitory effects on active infant immunization is an important factor hampering the use of certain vaccines, such as measles or respiratory syncytial virus vaccine, in early infancy. The resulting delay in protection by conventional or novel vaccines may have significant public health consequences. To define immunization approaches which may circumvent this phenomenon, experiments were set up to further elucidate its immunological bases. The influence of maternal antibodies on antibody and T cell responses to measles hemagglutinin (MV-HA) were analyzed following MV-HA immunization of pups born to immune or control BALB/c mothers using four different antigen delivery systems: live or inactivated conventional measles vaccine, a live recombinant canarypox vector and a DNA vaccine. High levels (> 5 log10) of maternal anti-HA antibodies totally inhibited antibody responses to each of the vaccine constructs, whereas normal antibody responses were elicited in presence of lower titers of maternal antibodies. However, even high titers of maternal antibodies affected neither the induction of vaccine specific Th1/Th2 responses, as assessed by proliferation and levels of IFN-gamma and IL-5 production, nor CTL responses in infant mice. On the basis of these unaltered T cell responses, very early priming and boosting (at 1 and 3 weeks of age, respectively) with live measles vaccine allowed to circumvent maternal antibody inhibition of antibody responses in pups of immune mothers. This was confirmed in another immunization model (tetanus toxoid). It suggests that effective vaccine responses may be obtained earlier in presence of maternal antibodies through the use of appropriate immunization strategies using conventional or novel vaccines for early priming. PMID- 9862351 TI - Similar as well as distinct MHC class I-binding peptides are generated by exogenous and endogenous processing of hepatitis B virus surface antigen. AB - Murine MHC class I-restricted cytotoxic T lymphocyte (CTL) responses can be primed by exogenous as well as endogenous hepatitis B surface antigen (HBsAg). Immunodominant CTL-defined epitopes of this viral envelope protein are the Ld binding 12-mer S28-39 peptide IPQSLDSWWTSL in H-2d mice, and the Kb-binding 8-mer S208-215 peptide ILSPFLPL in H-2b mice. We tested if CTL recognizing these epitopes can be primed in vivo by HBsAg delivered as either an exogenous antigen (native HBsAg lipoprotein particles), or an endogenous antigen (plasmid DNA encoding HBsAg). Primed T cells were restimulated in vitro prior to the cytotoxicity assay with cells presenting the H-2 class I-binding epitopes generated by either exogenous or endogenous processing of HBsAg. The data indicate that the Ld-binding peptide S28-39 is generated during exogenous as well as endogenous processing of HBsAg. In contrast, the Kb-binding peptide S208-215 is generated during exogenous but not endogenous processing of HBsAg. Hence, some but not all MHC class I-binding, immunogenic peptides are generated during endogenous and exogenous processing of HBsAg but there also exists a repertoire of immunogenic peptides of viral origin that is only revealed after exogenous processing of viral proteins. PMID- 9862352 TI - Delayed kinetics of tumor necrosis factor-mediated bystander lysis by peptide specific CD8+ cytotoxic T lymphocytes. AB - Mouse CD8+ CTL reactive with an H-2Db presented 9-mer peptide of the human papilloma virus 16 (HPV-16) protein E749-57 (RAHYNIVTF) were generated from the splenocytes of wild-type C57BL/6 (B6), B6.perforin-deficient, B6.gld or B6.TNF deficient mice. In short-term (4 h) assays, CTL from B6, B6.TNF-deficient and B6.gld mice displayed peptide-specific perforin- and/or Fas ligand (FasL) mediated lysis of E7-transfected mouse RMA lymphoma cells (RMA-E7) or E749-57 peptide-pulsed RMA-S cells, while CD8+ CTL from B6.perforin-deficient mice lysed via FasL exclusively. Rapid and efficient lysis of syngeneic bystander B6 spleen T cell blasts by B6, B6.TNF-deficient or B6.perforin-deficient antigen-activated CTL was mediated apparently exclusively by a FasL/Fas mechanism. By contrast CTL from B6.gld mice did not mediate rapid bystander lysis of B6 blasts. Rather B6.gld CTL delivered delayed bystander lysis after 36-48 h that was mediated by TNF. TNF-mediated bystander lysis of syngeneic blasts appeared to be independent of class I molecules and was mediated at least in part by soluble TNF. By contrast, there was no evidence that soluble FasL-mediated bystander lysis. For the first time, these data indicate that CD8+ CTL may use FasL or TNF in a kinetically and physically distinct fashion to mediate bystander killing. PMID- 9862353 TI - Ligation of CD5 on resting B cells, but not on resting T cells, results in apoptosis. AB - The CD5 molecule is expressed by a B cell subset. We have demonstrated that resting B cells do not proliferate in response to CD5 ligation, whereas cells preactivated with anti-IgM and IL-2 do so. Here, we specifically studied the effects of anti-CD5 and anti-IgM on apoptosis of CD5+ B cells. Both ligation of CD5 or of surface IgM (sIgM) resulted in apoptosis. This started earlier following ligation of CD5 than with sIgM, and both responses were time dependent. CD5-induced apoptosis was independent of the epitope recognized or the way the antibody was presented to the B cells. CD5+ B cells were more sensitive to IgM induced apoptosis than CD5 B cells. Engagement of CD5 or CD3 expressed by T cells failed to induce apoptosis. Our data indicate differences in the function of CD5 molecules on tonsillar B cells, compared with blood T cells and suggest that cross-linking CD5 on B cell activates specific pathways responsible for apoptosis. PMID- 9862354 TI - Characterization of T cell-expressed chimeric receptors with antibody-type specificity for the CD4 binding site of HIV-1 gp120. AB - Chimeric T cell receptors (cTCR) with an antibody specificity have been proposed in several models as a combination of antibody and cellular immunotherapy without MHC restriction. Such a tool could be of a limited use in HIV infection because of the great variability of the virus. The human single-chain antibody (ScFv-b12) derives from the b12 antibody directed to the CD4 binding site of gp120, a potent neutralizer of different HIV-1 strains, including a large panel of primary isolates. A single-chain fragment variable (ScFv) bearing the VH Pro-->Glu mutation that improves b12 affinity 54-fold, called ScFv-b12E, was also constructed. The ScFv were linked to the signal-transducing y chain of the Fc(gamma)RIII, with or without spacer region, and expressed in the murine MD45 T cell line. The different cTCR formats behave similarly in terms of ScFv surface expression, but differ according to their activation threshold. T cell transfectants can be stimulated with immobilized gp120 derived from all HIV strains tested. BHK cells infected with Semliki forest virus (SFV) carrying an HIV-1 envelope gene (SFV-env) derived from either HIV-1 laboratory strains (LAI, MN12, HXB2) or field isolates (BX08, CHAR or 133) were used as targets for the transfectants. All gp120-expressing cells induced cTCR-specific activation. The latter result is contrasting with the lack of specific recognition of SFV-CHAR- or 133-infected cells by the native b12 antibody, as measured by cytofluorometric analysis. Finally, HeLa cells (which constitutively express the coreceptor CXCR4) are able to bind HIV-1 gp160 when transfected with the chimeric receptor ScFv-b12 gamma, but, importantly, do not become infected by the virus. Our results therefore suggest that cTCR with b12 specificity can confer to T cells broad anti HIV reactivity without making them susceptible to HIV infection. PMID- 9862355 TI - Production of reactive oxygen intermediates following CD40 ligation correlates with c-Jun N-terminal kinase activation and IL-6 secretion in murine B lymphocytes. AB - Recent studies indicate that reactive oxygen intermediates (ROI) serve as second messengers in cell signaling. ROI have been implicated in the activation of NF kappaB as well as c-Jun N-terminal kinase (JNK) in response to IL-1 and TNF-alpha stimulation. In this report we examine whether intracellular ROI are involved in CD40 receptor signaling. We show that CD40 engagement on resting splenic B lymphocytes and murine B lymphoma WEHI 231 cells generates ROI. Blocking ROI production by preincubation with the antioxidant N-acetyl-L-cysteine inhibits JNK activation, NF-kappaB-driven luciferase activity, and IL-6 secretion following CD40 ligation, suggesting a role for ROI in CD40-mediated signaling events. Furthermore, transfection of WEHI 231 cells with a plasmid encoding Mn-superoxide dismutase interferes with CD40-induced NF-kappaB activation, providing further support for ROI involvement in this pathway. Collectively, these data demonstrate that ROI may serve as second messengers linking CD40 engagement on B cells to important downstream activation events. PMID- 9862356 TI - Alpha1/alpha2 domains of H-2D(d), but not H-2L(d), induce "missing self" reactivity in vivo--no effect of H-2L(d) on protection against NK cells expressing the inhibitory receptor Ly49G2. AB - Introduction of the MHC class I transgene H-2Dd on C57BL/6 (B6) background conveys NK cell-mediated "missing self" reactivity against transgene-negative cells, and down-regulates expression of the inhibitory receptors Ly49A and Ly49G2 in NK cells. We here present an analysis of transgenic mice expressing chimeric H 2Dd/Ld MHC class I transgenes, and show that the alpha1/alpha2 domains of H-2Dd were necessary and sufficient to induce "missing self" recognition and to down modulate Ly49A and Ly49G2 receptors. In contrast, transgenes containing the alpha1/alpha2 domains of H-2Ld induced none of these changes, suggesting that not all MHC class I alleles in a host necessarily take part in NK cell education. The lack of effect of the alpha1/alpha2 domains of H-2Ld on NK cell specificity was surprising, considering that both H-2Ld and H-2Dd have been reported to interact with Ly49G2. Therefore, the role of H-2Ld for protection against NK cells expressing Ly49G2 was re-investigated in a transfection system. In contradiction to earlier reports, we show that H-2Dd, but not H-2Ld, abolished killing by sorted Ly49G2+ NK cells, indicating that H-2Ld does not inhibit NK cells via the Ly49G2 receptor. PMID- 9862357 TI - Activation of nuclear factor-kappaB in macrophages by mycoplasmal lipopeptides. AB - Mycoplasmas are potent macrophage stimulators. The active principle are lipopeptides or lipoproteins with a characteristic N-terminal S-[dihydroxypropyl] cysteinyl group bearing two ester-bound fatty acids and lacking the amide-bound one common to other bacterial lipoproteins. Using synthetic analogues of mycoplasmal lipopeptides, we investigated activation of the transcription factor NF-kappaB in the C3H/HeJ mouse-derived DMBM-3 cell line. The lipopeptides activated NF-kappaB at below nanomolar concentrations. Activation in the murine system occurred distinctly earlier than TNF-alpha liberation, excluding autocrine stimulation by TNF-alpha. As determined from a supershift experiment, the active NF-kappaB complex consisted of the heterodimer p50/p65(RelA). The relevance of these findings for the inflammatory response to mycoplasmas and for mycoplasma mediated effects on HIV-infected macrophages is discussed. PMID- 9862358 TI - CTLA4 (CD152) modulates the Th subset response and alters the course of experimental Leishmania major infection. AB - Since both the nature and the amplitude of an antigen-specific T cell response are dependent on co-stimulatory signals, we have investigated the role of CD28/CD152-mediated T cell co-stimulation in the regulation of experimental cutaneous leishmaniasis. CD28-deficient mice and their wild-type littermates are equally susceptible to Leishmania major infection. Whole anti-CD152 antibody significantly exacerbates the disease while anti-CD152 Fab ameliorates the disease in genetically susceptible BALB/c mice but not in C57BL/6, a resistant strain. The anti-CD152-induced exacerbation of the disease is accompanied by increased IL-4-secreting cell number, diminished parasite-specific delayed-type hypersensitivity (DTH) response and augmented anti-2,4,6-trinitrophenyl (TNP) IgG1 in response to TNP-leishmanial antigen crude soluble antigen (CSA), suggesting an exaggerated Th2 type of response. Anti-CD152 Fab-mediated amelioration of the disease is associated with increased IFN-gamma-secreting cell number, increased parasite-specific DTH response and enhanced IgG2a isotype in response to TNP-CSA suggesting a Th1 type of response. Unlike TNP-CSA, TNP keyhole limpet hemocyanin does not induce the change in Ig isotype, indicating that the immunomodulatory effect of anti-CD152 is antigen specific. Anti-CD152 antibody-induced early change in Th subsets suggests an important role for CD152 in determining the course of L. major infection, perhaps by alteration of Th subset differentiation. PMID- 9862359 TI - Distinct roles for B7 costimulation in contact hypersensitivity and humoral immune responses to epicutaneous antigen. AB - Productive interactions between B7-1 and B7-2 costimulatory molecules on dendritic cells (DC) and CD28 on T cells are thought to be critical for successful antigen presentation. Epicutaneous application of haptens induces both contact hypersensitivity (CHS), an inflammatory cutaneous response mediated by CD8+ T cells, and an anti-hapten antibody response mediated by CD4+ helper T cells. The role of B7 costimulation in the immune response to oxazolone (Ox) was analyzed using mice lacking either B7-1 (B7-1-/-), B7-2 (B7-2-/-), or both (Db-/ ) of these costimulatory molecules. The absence of both B7-1 and B7-2 results in diminished CHS. This inhibition is largely overcome at higher hapten sensitizing doses indicating the presence of compensatory pathways. In contrast, anti-Ox IgG1 and IgG2a responses were not detected in the absence of both B7-1 and B7-2, even at high sensitizing doses, indicating an obligatory role of B7 costimulation in IgG class switching. B7-1 and B7-2 have overlapping functions in both CHS responses and anti-hapten response. B7-2-/- mice demonstrated a modestly reduced CHS response only at very low doses of Ox (0.05%), but responded normally at higher Ox doses, and B7-1-/- mice had CHS responses indistinguishable from those of wild-type mice. Similarly, anti-Ox IgG responses were comparable in wild-type, B7-1-/- and B7-2-/- mice. Taken together, these studies reveal distinct roles for B7 costimulation in response to epicutaneous antigens with an obligatory role for IgG class switching and an important, but nonessential role for CHS responses. PMID- 9862360 TI - Accessibility changes across the mouse Igh-V locus during B cell development. AB - During their development, B and T lymphocytes are thought to undergo several cycles of chromatin remodeling at their antigen receptor loci that serve to regulate access of a common V(D)J recombinase to particular gene segments. We used germ-line transcription and susceptibility to DNasel as markers to examine tissue and stage-specific changes in chromatin structure surrounding genes of the VHJ558, VH10, and VHS107 families, whose members are located at discreet subregions of the locus. Germ-line VH transcripts from all three families were detectable at pro- and pre-B cell stages. Transcripts from the VH10 and VHS107 families, but not VHJ558, remained detectable at the immature and mature B cell stages. Unexpectedly, none of the germ-line VH loci examined were markedly nuclease sensitive, regardless of cell type or transcriptional activity. A modest degree of nuclease sensitivity was noted at the VHJ558 loci of pro-B and pre-B cells, however. Our data suggest that the entire Igh-V locus becomes accessible at early B cell stages, and returns thereafter to an inaccessible state. However, the timing of these accessibility changes does not occur uniformly across the VH array. These results imply that multiple long-range elements are involved in targeting VH genes for rearrangement. PMID- 9862362 TI - Deficient interleukin-10 production by neonatal T cells does not explain their ineffectiveness at promoting neonatal B cell differentiation. AB - Neonatal T cells are poor promoters of Ig secretion by neonatal B cells. Since IL 10 has been shown to play a role in B cell differentiation, we investigated the relationship of IL-10 production by neonatal T cells and their ability to provide B cell help. Neonatal CD4+(CD8-) T cells and adult naive CD4+ (CD8-/CD45RO-) T cells activated with immobilized anti-CD3 produced consistently less IL-10 than adult memory CD4+(CD8-/CD45RA-) T cells. Production of IL-10 by adult and neonatal T cells was dependent on IL-2, but was unaffected by supplemental IL-4. Despite diminished IL-10 production, supplemental IL-10 increased neonatal T cell dependent Ig secretion only modestly, but did not increase Ig heavy chain isotype switching. This contrasted with the ability of IL-10 to enhance the secretion of all Ig isotypes by adult B cells stimulated in the presence of either IL-2 or IL 4. These results suggest that IL-10 can promote T cell-dependent Ig secretion but not Ig heavy chain isotype switching by neonatal B cells. However, deficient IL 10 production alone does not account for the poor ability of neonatal T cells to support neonatal B cell Ig production. PMID- 9862361 TI - Genetic origin of IgG antibodies cloned by phage display and anti-idiotypic panning from three patients with autoimmune thrombocytopenia. AB - The beneficial use of intravenous immunoglobulins (IVIG) in certain groups of patients with autoimmune thrombocytopenic purpura (AITP) has been proven. AITP is a severe disease in children with a still unknown etiology. It is not clear how IVIG functions in this and other autoimmune diseases. To analyze and compare patient-derived monoclonal IgG antibodies that are bound by IVIG in an anti idiotypic manner, the combinatorial antibody phage display system was applied. From three different patients with AITP, a large number of clones specifically reacting with IVIG molecules were enriched. The heavy and light chain variable regions were sequenced and compared with each other and with databases. Many variable regions showed extensive replacement mutations within the complementarity-determining regions, while two were identical to germ-line genes. Our data show that the most frequently used germ-line gene loci of these IVIG binders are identical to those observed for many other autoantibodies. This implicates a specific interaction of IVIG particularly with autoantibodies and B cell receptors derived from germ-line genes that are often used for the generation of autoantibodies. PMID- 9862363 TI - Linkage of a major quantitative trait locus to Yaa gene-induced lupus-like nephritis in (NZW x C57BL/6)F1 mice. AB - In the present study, we mapped the major quantitative trait loci (QTL) differing between the NZW and C57BL/6 inbred strains of mice by making use of (NZW x C57BL/6.Yaa)F1 mice, a model in which the lupus-like autoimmune syndrome observed in male mice is associated with the presence of an as yet unidentified Y chromosome-linked autoimmune acceleration gene, Yaa. Linkage analysis of 126 C57BL/6 x (NZW x C57BL/6.Yaa)F1 backcross males provided evidence for a major QTL on chromosome 7 controlling both the severity of glomerulonephritis and the production of IgG anti-DNA autoantibody and retroviral gp70-anti-gp70 immune complexes. Two additional QTL of C57BL/6 origin on chromosome 17 had no apparent individual effects, but showed strong epistatic interaction with chromosome 7 QTL for disease severity and anti-DNA autoantibody production. Our data also identified on chromosome 13 a QTL of NZW origin with a major effect on the level of gp70, and showing an additive effect with the chromosome 7 QTL on the level of gp70 immune complexes. Our study thus provides a model to dissect the complex genetic interactions that result in manifestations of murine lupus-like disease. PMID- 9862364 TI - Soluble CD21 induces activation and differentiation of human monocytes through binding to membrane CD23. AB - Interactions between CD23, the low-affinity receptor for IgE, and CD21, the C3d/EBV receptor, modulate several intracellular events in lymphocytes. A soluble form of CD21 (sCD21) corresponding to the extracellular domain of the receptor circulates in normal plasma. We now demonstrate that purified sCD21 acts as a functional ligand for CD23-expressing monocytes. Soluble CD21 induced an increase in intracellular cGMP levels and the production of IL-6 and TNF-alpha in IL-4 pretreated monocytes induced to express CD23 but not in unstimulated CD23- monocytes. The accumulation of cGMP and the production of TNF-alpha were inhibited by NG-monomethyl-L-arginine (L-NMMA), indicating that sCD21 activates the L-arginine pathway of NO production. We demonstrated that sCD21 activates NO synthase (NOS) since it was found to enhance the conversion of L-arginine into L citrulline and induce the intracellular expression of inducible NOS in CD23+ monocytes. In addition, sCD21 was shown to up-regulate the expression of HLA-DR and CD40 and decrease that of CD14 on cultured CD23+ monocytes. Thus, in a fashion similar to IgE complexes, sCD21 is able to efficiently trigger CD23 signaling pathways, inducing the release of pro-inflammatory mediators by human monocytes. Soluble CD21 up-regulates the expression of molecules involved in antigen presentation, further suggesting a potential immunoregulatory function for the soluble molecule. PMID- 9862365 TI - Intestinal lymphangiectasia, a disease characterized by selective loss of naive CD45RA+ lymphocytes into the gastrointestinal tract. AB - Intestinal lymphangiectasia (InL) is a disease characterized by hypoproteinemia and lymphocytopenia resulting from blocked intestinal lymphatics and loss of lymph fluid into the gastrointestinal tract. This leads to immunologic abnormalities including hypogammaglobulinemia, skin anergy and impaired allograft rejection. In the present study, we evaluated whether the above immunologic abnormalities are secondary to a quantitative or qualitative disorder of T cells. In initial studies we demonstrated that adult InL patients' peripheral blood contain strikingly (and significantly) reduced numbers of CD4+/CD45RA+ T cells, whereas the numbers of CD4+/CD45RO+ T cells were only moderately (and not significantly) reduced. In addition, the CD4+/CD45RO+ T cell population contained an increased percentage of highly differentiated and previously sensitized cells, as demonstrated by decreased CD27 and CD31 expression and increased HLA-DR and CD69 expression. In subsequent functional studies, we showed that the InL CD4+/CD45RO+ T cells, when stimulated in vitro, proliferate fivefold less than control CD4+/CD45RO+ T cells and produce fourfold more IL-4 and threefold less IFN-gamma and IL-2. Thus, this cytokine production profile also reflects the highly differentiated nature of the residual cell population. Overall, these studies provide new information on the trafficking of naive/mature and Th1/Th2 T cell populations in this disease model. PMID- 9862366 TI - The distribution of IL-13 receptor alpha1 expression on B cells, T cells and monocytes and its regulation by IL-13 and IL-4. AB - To study the expression of IL-13 receptor alpha1 (IL-13Ralpha1), specific monoclonal antibodies (mAb) were generated. Surface expression of the IL 13Ralpha1 on B cells, monocytes and T cells was assessed by flow cytometry using these specific mAb. Among tonsillar B cells, the expression was the highest on the IgD+ CD38- B cell subpopulation which is believed to represent naive B cells. Expression was also detectable on a large fraction of the IgD-CD38- B cells but not on CD38+ B cells. Activation under conditions which promote B cell Ig class switching up-regulated the expression of the receptor. However, the same stimuli had an opposite effect for IL-13Ralpha1 expression levels on monocytes. While IL 13Ralpha1 mRNA was clearly detectable in T cell preparations, no surface expression was detected. However, permeabilization of the T cells showed a clear intracellular expression of the receptor. A soluble form of the receptor was immunoprecipitated from the supernatant of activated peripheral T cells, suggesting that T cell IL-13Ralpha1 might have functions unrelated to the capacity to form a type II IL-4/IL-13R with IL-4Ralpha. PMID- 9862367 TI - c-Rel is essential for B lymphocyte survival and cell cycle progression. AB - c-Rel is a lymphoid-specific member of the NF-kappaB/Rel family of transcriptional factors. To investigate the role of c-Rel in B lymphocyte function, we generated a c-Rel(-/-) mouse via a gene targeting approach. Although early lymphocyte development is normal in c-Rel(-/-) mice, there are significantly fewer B cells displaying a memory (IgM/IgD-) phenotype. Upon immunization, c-Rel(-/-) mice generate fewer B cells with a germinal center (PNAhi) phenotype. In vitro, c-Rel(-/-) B cells proliferate poorly upon ligation of their surface IgM or CD40 receptors or when stimulated with either lipopolysaccharide (LPS) or T cell help. Early molecular events that precede proliferation, such as increases in RNA synthesis as well as IL-2 receptor alpha chain expression, are greatly diminished in c-Rel(-/-) B cells. Furthermore, c Rel(-/-) B cells are impaired in the ability to receive survival signals generated by anti-IgM or LPS. In contrast, CD40-mediated cell survival is normal in c-Rel(-/-) B cells, suggesting the involvement of a survival-signaling pathway that is independent of c-Rel. When c-Rel (-/-) B cells are co-stimulated with either anti-IgM and CD40 or LPS and CD40, they are rendered capable of progressing through the cell cycle. Finally, co-culture experiments suggest that the defects observed in c-Rel(-/-) B cells are intrinsic to the cell and can not be rescued through either cell-cell contact or addition of soluble factors. Thus, c-Rel is requisite for differentiation to the germinal center and memory B cells in vivo and is required for the transduction of survival and cell cycle progression signals mediated by anti-IgM and LPS in vitro. Furthermore, while c Rel is involved in CD40-induced proliferation, it is apparently dispensable for the survival signals transduced by CD40. PMID- 9862368 TI - Deaggregated homologous immunoglobulin-peptide conjugates induce peptide-specific T cell nonresponsiveness in vivo. AB - Previous studies have proven the efficacy of intravenous injection of deaggregated protein as a means of inducing tolerance. In the present study, the immunodominant peptide 70-86 of myelin basic protein (MBP) was covalently linked to either mouse Ig or Lewis rat IgG. Lewis rats immunized with MBP in complete Freund's adjuvant were completely protected from development of experimental allergic encephalomyelitis (EAE) by their injection with as little as 40 microg of peptide conjugate on days 0 and 10 after immunization. Peptide-specific proliferative and cytokine responses by T cells from treated rats in vitro were severely depressed compared with controls, while responses to whole MBP were unaffected. Significantly, injections of 100 microg of peptide conjugate on days 0 and 4 after adoptive transfer of peptide-specific T lines protected rats from passive EAE while a single injection of 100 microg of conjugate at the onset of active EAE prevented any further disease progression. Both results suggest that primed effector cells as well as naive T cells are prone to tolerance induction by this means. The ability to intervene in ongoing immune responses with such specificity may be useful therapeutically in control of autoimmunity or allergic responses to environmental antigens. PMID- 9862369 TI - Enhanced murine CD4+ T cell responses induced by the CD2 ligand CD48. AB - The physiological functions of murine CD2 and its ligand CD48 are uncertain. We have examined the role of the CD2-CD48 interaction in murine T cell activation using a series of Chinese hamster ovary (CHO) cell transfectants. CHO cells expressing I-Ad together with CD48 induced more potent activation of OVA specific, I-Ad-restricted DO11.10-transgenic T cells than CHO cells expressing I Ad alone. CD48 augmented proliferation and IL-2 production in response to antigen. The enhancing effect of CD48 was of the same magnitude as that seen for CD80 (B7-1). Conjugate assays revealed the ability of CD48 to increase adhesion between T cells and CHO transfectants. The enhancing effects of CD48 on T cell antigen-presenting cell adhesion and T cell activation were inhibited by anti-CD2 monoclonal antibody. This report provides the first evidence that the CD2 ligand CD48 contributes to the interactions of murine CD4+ T cells with antigen presenting cells. PMID- 9862370 TI - Signaling through the tetraspanin CD82 triggers its association with the cytoskeleton leading to sustained morphological changes and T cell activation. AB - In this report, we provide new evidence of a crosstalk between T cell activation and adhesion processes through a functional cytokeleton. We show that CD82 signaling induces long-lasting adhesion, spreading and development of membrane extensions, involving actin polymerization. Addition of various co-stimuli (phorbol 12-myristate 13-acetate or monoclonal antibodies to CD3 or CD2) increases the CD82-induced morphological alterations and, reciprocally, CD82 engagement synergizes with these stimuli to induce T cell activation as indicated by both primary tyrosine phosphorylation and IL-2 production. Different kinases are involved in both processes. CD82 co-signaling involves src kinases including p56 Ick. On the other hand, the CD82-induced alterations of cell morphology are negatively regulated by cAMP-dependent kinases independently of activation of src kinases. Simultaneously with cytoskeletal rearrangements, we observed an inducible association of CD82 with the cytoskeletal matrix. In addition, the potentiating and stabilizing effects induced by CD82 cross-linking on tyrosine phosphorylation were abolished by cytoskeleton-disrupting agents. These results suggest that the actin polymerization triggered by CD82, through its ability to associate with the cytoskeletal matrix, is the primary step involved in the CD82 induced co-stimulatory activity. Our data provide further evidence for a direct role of the actin cytoskeleton as a major component for sustained signal transduction in T cells and suggest that tetraspanins could be "membrane organizers" connecting both surface and intracellular molecules. PMID- 9862371 TI - Protection from Plasmodium berghei infection by priming and boosting T cells to a single class I-restricted epitope with recombinant carriers suitable for human use. AB - The desirability of inducing cytotoxic T cell responses to defined epitopes in humans has led to the development of a variety of recombinant delivery systems. Recombinant protein particles derived from a yeast retrotransposon (Ty) and the modified Ankara vaccinia (MVA) virus can deliver large epitope strings or even whole proteins. Both have previously been administered safely in humans. Immunization with recombinant Ty and MVA containing a single Plasmodium berghei class I-binding epitope provided 95% sterile protection against malaria in mice. The sequence of immunization, Ty followed by MVA, was critical to elicit high levels of IFN-gamma-producing cells and protection. The reciprocal sequence (MVA/TY) or homologous boosting was not protective. Both constructs (Ty and MVA) contain the H-2Kd-restricted pb9 CTL epitope from the circumsporozoite protein of P. berghei among a string of 8-15 human P. falciparum-derived CTL epitopes restricted through 7 common HLA alleles as well as widely recognized CD4 T cell epitopes. Thus, the novel recombinant Ty/MVA prime/boost combination with these constructs provides a safe alternative for evaluation for human vaccination against P. falciparum malaria. PMID- 9862373 TI - Thymus-dependent monoclonal antibody-induced protection from transferred diabetes. AB - It is well established that long-term protection from insulin-dependent diabetes mellitus (IDDM) can be afforded to non-obese diabetic (NOD) mice by a short course of non-depleting (nd) anti-CD4 monoclonal antibodies (mAb). Since it is increasingly apparent that the CD8+ T cell plays a prominent role in the development of IDDM, we have investigated the effect of an anti-CD8 mAb (YTS 105) of the same isotype in both spontaneous and induced IDDM in NOD mice. Treatment with YTS 105 for 3 weeks was able to prevent the transfer of IDDM for a long period, and also substantially reduced spontaneous IDDM in female NOD mice. The role of the thymus in tolerance induction by these antibodies was studied. In the adult transfer model, thymectomized NOD mice, unlike their euthymic counterparts, were not protected long-term by treatment with YTS 105, and began to become overtly diabetic shortly after treatment. This was also true when the nd anti-CD4 mAb was used. Protection from spontaneous disease was not affected in the same way by thymectomy. The reasons for the observed effect of the thymus in the transfer model, and the differences between the two models that may explain the contrasting results are discussed. PMID- 9862372 TI - The Qa-1b molecule binds to a large subpopulation of murine NK cells. AB - Recent studies on human NK cells have demonstrated that the NK cell CD94/NKG2 receptors bind to the nonclassical MHC class I molecule HLA-E. A functional CD94/NKG2 complex has not yet been identified in rodents, but cDNA encoding rat and mouse CD94 and NKG2 have recently been cloned, suggesting that CD94/NKG2 receptors may exist in species other than man. The mouse nonclassical MHC class I molecule Qa-1 shares several features with HLA-E. This suggests that Qa-1 may be similarly recognized by murine NK cells. To study the ability of Qa-1 to bind to murine NK cells, we have produced a soluble tetrameric form of Qa-1b. In the present study, we demonstrate that Qa-1b tetramers distinctly bind to a large subset of fresh or IL-2-activated NK1.1+/CD3- splenocytes independently of the expression of Ly49 inhibitory receptors. Binding occurs whether NK cells have evolved in an MHC class I-expressing or in an MHC class I-deficient environment. Our data suggest the existence of a Qa-1-recognizing structure on a large subpopulation of murine NK cells that may be similar to the human CD94/NKG2 heterodimeric complex. PMID- 9862374 TI - Responses to human rhinovirus in CD45 T cell subsets isolated from tonsil. AB - Isotypes of CD45 have been used extensively as markers of memory and naive populations of T cells in peripheral blood. In this study, T cells were isolated from human tonsil and their proliferative response against human rhinovirus was measured. Unexpectedly, equivalent responses were found among the CD4+CD45RA+ and CD4+CD45RO+ populations of T cells. This response requires MHC class II-positive antigen-presenting cells. The time course of the T cell response in vitro was that of a classical recall response, and no proliferative response to the virus could be detected in human cord blood. These results suggest that tonsils contain a significant population of CD45RA+ memory cells. The presence of this population may reflect ongoing stimulation with this common infectious agent, and the anatomical location of the T cells within the major lymphoid organ draining the naso-pharyngeal epithelial surface. PMID- 9862375 TI - T cell selection during the evolution of CD8+ T cell memory in vivo. AB - Memory T cell responses are frequently highly restricted in terms of receptor usage. How and when such clonotypic dominance is established remains poorly understood. Here we have investigated the evolution of the T cell responses to an epitope from Epstein-Barr virus (EBV), (FLRGRAYGL), by analyzing TCR use of clones specific for this epitope, derived from peripheral blood mononuclear cells taken from individuals early during primary EBV infection and up to 3 years later. We show that, in a given individual, particular T cell clonotypes are selected early during the primary response to this epitope and that the same clonotypes dominate the late memory response. In one individual direct analysis of HLA-B8-restricted FLRGRAYGL-specific T cells, isolated from peripheral blood lymphocytes taken during primary EBV infection using a tetrameric MHC-peptide complex, confirmed the early selection of the dominant clonotypes. PMID- 9862376 TI - Diverse CD1d-restricted reactivity patterns of human T cells bearing "invariant" AV24BV11 TCR. AB - Human and murine natural T (NT) cells, also referred to as NK1.1+ or NK T cells, express TCR with homologous V regions (hAV24/BV11 and mAV14/BV8, respectively) and conserved "invariant" TCR AVAJ junctional sequences, suggesting recognition of closely related antigens. Murine NT cells recognize CD1-expressing cells and are activated in a CD1-restricted fashion by several synthetic alpha glycosylceramides, such as alpha-GalCer. Here we studied the reactivity of human T cells against CD1d+ cells pulsed or not with alpha-GalCer and other related ceramides. CD1d-restricted recognition of alpha-GalCer was a general and specific feature of T cell clones expressing both BV11 and canonical AV24AJ18 TCR chains. Besides, human and murine NT cells showed the same reactivity patterns against a set of related glycosylceramides, suggesting a highly conserved mode of recognition of these antigens in humans and rodents. We also identified several AV24BV11 T cell clones self reactive against CD1+ cells of both hemopoietic and nonhemopoietic origin, suggesting the existence of distinct NT cell subsets differing by their ability to recognize self CD1d molecules. PMID- 9862377 TI - TNF-alpha and IFN-gamma render microglia sensitive to Fas ligand-induced apoptosis by induction of Fas expression and down-regulation of Bcl-2 and Bcl-xL. AB - The immune response in the central nervous system (CNS) involves microglial cells which represent intraparenchymal antigen-presenting cells (APC). To control immune effector mechanisms it may be required to induce apoptosis of APC and thereby limit reactivation of T cells that have invaded the CNS. In the present study we investigated the susceptibility of primary murine microglia and of the murine microglial cell line BV-2 to undergo Fas-mediated apoptosis. Whereas resting microglia are resistant to Fas ligand (FasL) treatment, induction of FasL mediated apoptosis was achieved by treatment with TNF-alpha or IFN-gamma. The effect of these cytokines was paralleled by up-regulation of Fas expression and down-regulation of Bcl-2 and Bcl-xL but not Bax. Activation of microglia by TNF alpha and IFN-gamma was also accompanied by increased amounts of mRNA for the apoptosis inhibitor FLIP, an effect which did not protect the cells from FasL induced apoptosis. The FasL-induced cell death pathway in microglia involves reactive oxygen intermediates because the antioxidants N-acetylcysteine and glutathione interfere with induction of apoptosis. Surprisingly, microglia constitutively express FasL on the cell surface. However, blocking of endogenous Fas-FasL interaction with Fas-Fc fusion protein did not enhance the survival of microglia, excluding the possibility of suicide or fratricide mechanisms. By their expression of FasL and their TNF-alpha/IFN-gamma-dependent sensitivity to the pro-apoptotic effect of exogenous FasL, microglial cells may influence the course of T cell-mediated diseases of the CNS. PMID- 9862379 TI - Quiescent memory B cells in human peripheral blood co-express bcl-2 and bcl-x(L) anti-apoptotic proteins at high levels. AB - The anti-apoptotic proteins bcl-2 and bcl-xL seem to exhibit strictly opposite expression patterns in normal lymphoid cell differentiation stages, with bcl-2 low and bxl-xL high in immature and mature proliferating cells, the reverse being the case in recirculating quiescent cells. However, it is in fact not known whether recirculating memory cells are bcl-xL low or high. We analyzed memory (immunoglobulin isotype-switched) B cells in human peripheral blood, which were small lymphocytes in the G0 phase of the cell cycle, but proliferated better than naive B cells in response to Staphylococcus aureus Cowan I. Ex vivo these cells co-expressed bcl-2 together with bcl-xL mRNA and protein at high levels. The mcl 1 mRNA level was low. The bcl-xL mRNA level decreased during culture in medium containing fetal calf serum, which implies that it is maintained in vivo by continuous or frequent, non-mitogenic signal(s). The high bcl-xL expression of memory B cells may be relevant with regard to their longevity and/or their capacity to undergo an accelerated secondary type immune response. PMID- 9862378 TI - 2F1 antigen, the mouse homolog of the rat "mast cell function-associated antigen", is a lectin-like type II transmembrane receptor expressed by natural killer cells. AB - Inhibitory lectin-like receptors expressed on the surface of hematopoietic cells are critically involved in regulation of their effector functions. Here we report that a novel mAb specific for mouse NK cells, 2F1, recognizes the mouse homolog of the mast cell function-associated antigen (MAFA), an inhibitory lectin-like transmembrane receptor expressed on rat mast cells. The 2F1 antigen (2F1-Ag) and rat MAFA are structurally highly conserved and contain a cytoplasmic motif similar to the immunoreceptor tyrosine-based inhibitory motif that is presumably utilized for inhibitory signaling. We also identified a human homolog that is closely related to the rodent MAFA/2F1-Ag proteins. Like rat MAFA, 2F1-Ag is probably encoded by a single gene, which exhibits relatively little polymorphism. Strikingly, while rat MAFA is considered a mast cell antigen, we have been unable to detect cell surface expression of 2F1-Ag by mouse mast cell lines, bone marrow derived mast cells, or peritoneal mast cells. Furthermore, mouse bone marrow derived mast cells were devoid of 2F1-Ag mRNA. Instead, we find that approximately 40% of mouse NK cells express 2F1-Ag. Thus, MAFA/2F1-Ag may modulate immunological responses on at least two different cell types bridging the specific and innate immune system. PMID- 9862380 TI - Distribution of the interstitial cells of Cajal in the human anorectum. AB - The interstitial cells of Cajal are proposed to have a role in the control of gut motility. The aim of this study was to establish the distribution of interstitial cells of Cajal in the wall of the normal human anorectum. Interstitial cells of Cajal express the proto-oncogene c-kit. Interstitial cells of Cajal were identified in the colon by immunohistochemical staining, using a rabbit polyclonal anti-c-kit antibody. Anorectal tissue was obtained at surgical resection for carcinoma of the colorectum. Density of interstitial cells of Cajal was graded. Statistical analysis was performed using chi2 tests. In the longitudinal and circular muscle layers of the rectum interstitial cells of Cajal were seen in the bulk of the muscle layer. In the intermuscular plane interstitial cells of Cajal encased the myenteric plexus. Interstitial cells of Cajal were found at the inner margin of the circular muscle and in association with neural elements of the submuscular plexus. Within the internal anal sphincter interstitial cells of Cajal were infrequently scattered among the muscle fibres. The density of interstitial cells of Cajal in the internal anal sphincter was significantly lower than that observed in the circular muscle layer of the rectum (P = 0.014). In conclusion, interstitial cells of Cajal are evenly distributed in the layers of the muscularis propria of the rectum, but have a lower density in the internal anal sphincter. PMID- 9862382 TI - Changes in left ventricular contractility with the phase of respiration. AB - The end-systolic wall stress (sigma(es))-velocity of circumferential fiber shortening (V(cfsc)) relation was defined during the respiratory cycle, in order to obtain a totally noninvasive measure of left ventricular contractility. Eight young, healthy subjects were studied with echocardiography and calibrated carotid pulse tracings, while performing slow paced breathing. Left ventricular sigma(es) vs. V(efsc) relation was determined by fitting linear regression line to data points obtained at different times during the respiratory cycle. Data are given as mean+/-1SD. Left ventricular sigma(es) and V(efsc) exhibited small but significant changes during the respiratory cycle: sigma(es) was highest in late inspiration (56.9+/-4.8 g/cm2) and lowest in late expiration (49.2+/-3.7 g/cm2); inversely, V(cfsc) was lowest during late inspiration (1.18+/-0.17 circ/s) and highest during late expiration (1.34+/-0.20 circ/s). The relation was significant in each subject (r = -0.64+/-0.13) and remained inverse and significant, when it was determined separately for inspiration and expiration (r = -0.61+/-0.17 and 0.68+/-0.12, respectively). At identical end-systolic wall stress, the velocity of shortening was greater during inspiration then expiration, suggesting that contractility was reduced during the expiratory phase. The reduced expiratory contractility might reflect increased vagal influence on the ventricular myocardium. PMID- 9862381 TI - Inhibition of vagal vasodilatation by a selective neuropeptide Y Y2 receptor agonist in the bronchial circulation of anaesthetised dogs. AB - Neuropeptide Y (NPY) is both co-stored and co-released with noradrenaline from sympathetic nerve terminals. In the cardiovascular system, NPY acts on two main receptor subtypes. At postjunctional, or Y1 receptors, NPY can cause both direct vasoconstriction and the potentiation of various constrictor agents. NPY acting at the presynaptic, or Y2 receptor, inhibits the release of neurotransmitter from autonomic nerves. In the present paper, we have used both sympathetic stimulation and the selective NPY Y2 receptor agonist, N-acetyl [Leu28,Leu31] NPY24-36, to examine the role of NPY in the inhibition of vagally mediated vasodilatation in the bronchial circulation of the anaesthetised dog. Stimulation of the cardiac end of the cervical vagus nerve at 1 Hz for 15 s (1 ms, 70 V) increased bronchial vascular conductance by 45%. This increase in flow was abolished by atropine. Sympathetic stimulation for 2.5 min at 16 Hz (1 ms, 20 V) produced a significant (P < 0.05) and prolonged (9 min) inhibition of the subsequent parasympathetically evoked vasodilatation. Similarly, the NPY Y2 receptor agonist, N-acetyl [Leu28,Leu31] NPY24-36, produced a significant (P < 0.05) and prolonged (15 min) inhibition of parasympathetically evoked vasodilatation. When vagus was stimulated at 2.5 Hz for 30 s (1 ms, 70 V), an atropine-resistant, but capsaicin sensitive vasodilatation was observed. Neither sympathetic stimulation nor the NPY Y2 receptor agonist could be demonstrated to inhibit this vasodilatation. These results suggest that NPY can inhibit cholinergic parasympathetic vasodilatation in the bronchial circulation by an action on NPY Y2 receptors. PMID- 9862383 TI - Evidence that NMDA receptors mediate the responses of putative RVLM presympathetic neurons to vagal afferent stimulation in rats. AB - Experiments were performed in 25 urethane-anaesthetized male Sprague-Dawley rats. Forty-six RVLM neurons were identified as putative presympathetic neurons according to their arterial baroreceptor reflex related properties, i.e. they were inhibited by stimulation of the aortic nerve; silenced by elevation of blood pressure and had a cardiac cycle related rhythm of spontaneous discharge. Responses of these neurons to tripled square wave stimulation of vagal afferents was tested by means of peristimulus time histograms. In addition to a long lasting inhibition (I2), some neurons had one (P0) or two excitatory peaks (P1 and P2), and there was a short-lasting inhibition (I1) between P1 and P2. After microinjection of CPP (0.1 microl, 50 mM) into the NTS, the inhibitory responses were blocked, but the excitatory peaks were not affected; in the CVLM, CPP microinjection of the same dose had a similar effect on the responses elicited by vagal afferent stimulation in 15 of the 24 neurons tested. No detectable effects were observed in 9 neurons. However, intravenously administered ketamine attenuated or abolished these responses, either inhibitory or excitatory, in a dose dependent way. These results suggest: (1) an NMDA mechanism is involved in both the inhibitory and the excitatory responses. For the inhibitory responses, the involvement is both in the NTS and in the CVLM; for the excitatory responses, it is probably in the RVLM. (2) There may be a direct excitatory amino acid (EAA) pathway from the NTS to the RVLM. PMID- 9862384 TI - Presence of a non-NMDA glutamate receptor subtype in the sympathetic nervous system of neonatal swine. AB - For the first time, the GluR-1 subtype of AMPA receptor was identified in the sympathetic nervous system of neonatal swine, an animal model of human development and heart disease. The rationale was to seek evidence of a role ascribed to glutamate in cardiorespiratory regulation in the laboratory rat. The receptor was demonstrated with the avidin-biotin immunoperoxidase technique by using an affinity-purified polyclonal antibody judged to be specific to Glu-R1 in several species. Glu-R1 immunoreactivity was regionally distributed in the thoracic spinal gray, and present intracellularly in neurons and within the surrounding neuropil. Sympathetic preganglionic neurons in the intermediolateral cell column of upper and lower thoracic spinal segments were intensely labeled and surrounded by labeled neuropil. High concentrations of Glu-R1 distinguished laminae II: substantia gelatinosa and the outer region of lamina III. Laminae I and V of the dorsal horn but not IV contained immunolabeled neurons. Arrays of moderately immunoreactive perikarya extended from an intermediate zone of laminae VII to the central gray. Glia and perivascular processes were not labeled, confirming previous observations [Tachibana, M., Wenthold, R.J., Morioka, H., Petralia, R.S., 1994. Light and electron microscopic immunocytochemical localization of AMPA-selective glutamate receptors in the rat spinal cord. J. Comp. Neurol. 344, 431-454]. Neuronal staining patterns corroborated evidence in rats indicating a postsynaptic localization of Glu-R1 associated with plasma membranes and cytoplasmic organelles [Martin, L.J., Blackstone, C.D., Levey, A.I., Huganir, R.L., Price, D.L., 1993. AMPA glutamate receptor subunits are differentially distributed in rat brain. Neuroscience 53, 327-358.; Rubio, M.E., Wenthold, R.J., 1997. Glutamate receptors are selectively targeted to postsynaptic sites in neurons. Neuron 18, 939-950]. Our data predict a role for L glutamate in postnatal development of cardiorespiratory reflexes in swine. PMID- 9862385 TI - The role of N-type Ca2+ channels in regulating excitability of guinea-pig sympathetic neurones. AB - Ca2+ entry through voltage-gated channels activated during the action potential modifies neuronal excitability by activating several types of K+ channel. We have determined the effects of Ca2+ influx through N-type Ca2+ channels in sympathetic paravertebral neurones of the guinea-pig, using the specific antagonist, omega conotoxin GVIA. Blockade of large conductance (BK) Ca2(+)-activated K+ channels slowed action potential repolarization but did not affect the peak amplitude of the conductance (gKCal) underlying the afterhyperpolarization. Blockade of small conductance (SK) Ca2(+)-activated K+ channels decreased gKCal but did not affect action potential repolarization. Blockade of N-type Ca2+ channels slowed action potential repolarization and reduced the peak amplitude of gKCa1. We conclude that Ca2+ entry via N-type channels activates both BK and SK channels in guinea pig sympathetic neurones. This differs from our previous observations in rat sympathetic neurones. PMID- 9862386 TI - Regional differences in sympathetic neurotransmission to cutaneous arteries in the guinea-pig isolated ear. AB - The effects of sympathetic nerve stimulation on different cutaneous arteries were examined in arteries isolated from guinea-pig ears, by measuring membrane potential changes in smooth muscle cells in response to electrical field stimulation. Resting membrane potential (RMP) was similar in proximal (main ear artery) and distal (3rd or 4th branch order) cutaneous arteries (mean -71 mV). Single stimuli evoked excitatory junction potentials (EJPs) in all arteries. The EJPs in proximal arteries were twice the amplitude, and the time constant of EJP decay was almost half the value, compared with distal cutaneous arteries. EJP amplitude was reduced by > 90% by suramin (30 microM) or alpha,beta,methylene-ATP (alpha,beta,m-ATP)(1 microM) in all proximal, and most distal arteries. Residual responses in distal arteries were resistant to tetrodotoxin. The N-type calcium channel blocker, omega-conotoxin GVIA (30 nM), reduced EJP amplitude by 70-100% in both proximal and distal arteries. Successive EJPs evoked by trains of stimuli at 1 to 5 Hz were depressed in amplitude in proximal arteries, but showed facilitation in distal arteries. EJP depression in proximal arteries was reversed to facilitation by the alpha2-adrenoceptor antagonist, yohimbine (30 nM). Trains of stimuli delivered at 10-20 Hz produced summation of EJPs and active membrane responses in 30% of proximal arteries. Active responses were never detected in distal arteries. Slow depolarizations following the EJPs were detected in most arteries after trains of stimuli, and were abolished by prazosin (0.3 microM) or omega-conotoxin GVIA (30 nM). The density of the perivascular plexus of axons innervating proximal arteries, demonstrated with catecholamine fluorescence histochemistry, was twice that in distal cutaneous arteries. These regional differences in sympathetic neurotransmission suggest that cutaneous vasoconstriction in response to thermoregulatory stimuli, which occurs predominantly in distal cutaneous segments, is likely to be qualitatively different from cutaneous vasoconstriction of proximal arteries in response to other physiological stimuli. PMID- 9862387 TI - Differing haemodynamic and catecholamine responses to exercise in three groups with peripheralautonomic dysfunction: insulin-dependent diabetes mellitus, familial amyloid polyneuropathy and pure autonomic failure. AB - The haemodynamic and catecholamine responses to supine exercise, and the effect on standing blood pressure (BP), were studied in three groups with peripheral autonomic dysfunction; insulin-dependent diabetes mellitus (IDDM), familial amyloid polyneuropathy (FAP) and pure autonomic failure (PAF). Healthy normal subjects were studied as controls. With exercise, BP increased in controls, was unchanged in IDDM and FAP, and fell in PAF. Heart rate (HR) increased more in controls than IDDM, FAP or PAF. Cardiac index (CI) increased less in IDDM than controls, FAP or PAF. Systemic vascular resistance (SVR) fell similarly in controls and IDDM, with a greater fall in FAP and PAF. Plasma noradrenaline increased in controls and IDDM only; plasma adrenaline did not change and plasma dopamine was undetectable in all groups. On standing, BP was unchanged in controls; BP fell pre- and post-exercise in IDDM, FAP and PAF, with a significantly greater fall post-exercise in FAP and PAF. In conclusion, the haemodynamic responses to supine exercise and to standing after exercise differed in the three groups with peripheral autonomic dysfunction. These differences, and also the similarities, between different forms of peripheral autonomic dysfunction, may be of relevance to the clinical assessment and therapy of these patients. PMID- 9862388 TI - Effects of myocardial hypertrophy on neural reflexes controlling cardiovascular function. AB - There are clinical and experimental evidences that the cardiopulmonary reflex function is impaired in chronic hypertension, but it could be due to myocardial hypertrophy rather than to hypertension itself. To test this hypothesis we evaluated the Bezold-Jarisch reflex in experimental conditions of myocardial hypertrophy and arterial normotension. Adult male Wistar rats were subjected to myocardial hypertrophy (MHR) treating them with the beta-adrenoceptor agonist isoproterenol (0.3 mg/kg/day, s.c.) for 15 days and compared with vehicle injected control rats (CR). No significant changes in body weight (283+/-14 vs. 299+/-9 g), resting mean arterial pressure (104+/-4 vs. 110+3 mm Hg) or heart rate (330+/-11 vs. 358+/-18 bpm) were observed in MHR compared to CR. As expected, MHR showed left and right ventricular and left atrial hypertrophy when compared to CR. The bradycardia and hypotension that characterizes the Bezold Jarisch reflex, induced by the 5-HT3, agonist phenyldiguanide (1.5-24.0 microg/kg, i.v.), were significantly decreased in MHR compared to CR. Cardiac muscarinic responsiveness, which was assessed by electrical stimulation of the efferent vagus in anesthetized animals or by stimulation of muscarinic receptors in isolated hearts, was unchanged or increased, respectively, in MHR compared to CR. Additional studies showed that the baroreflex and chemoreflex were also attenuated in MHR compared to CR. These data indicate that cardiac hypertrophy impairs the Bezold-Jarisch reflex probably due to changes at central integrative areas of the reflex. PMID- 9862389 TI - Central somatostatin diminished inhibitory action of central CGRP on pancreatic basal secretion in conscious rats. AB - We examined whether central somatostatin prevents an inhibitory effect of central calcitonin-gene related peptide (CGRP) on pancreatic secretion in conscious male Wistar rats (330-330 g). Rats were prepared with separate cannulas for draining bile and pancreatic juice and with a duodenal cannula and an extrajugular vein cannula. In addition, another cannula was stereotactically implanted into the left lateral cerebral ventricle. Rats were placed in restraint cages and experiments were conducted 4 days after the operation without anesthesia. An injection of CGRP (0.1, 1.0 nmol/10 microl) into the left lateral cerebral ventricle (i.c.v.) inhibited pancreatic secretion dose-dependently. To confirm the inhibitory effect of CGRP (i.c.v.) was mediated via sympathetic nerves, phentolamine was injected intravenously (i.v.) bolus (0.5 mg kg(-1)) 0.5-h before CGRP (i.c.v.), followed by continuous infusion of 0.2 mg kg(-1) h(-1). Phentolamine (i.v.) reversed the inhibition produced by CGRP (i.c.v.). An injection of 4 nmol/10 microl somatostatin (i.c.v.) 5 min prior to CGRP injection diminished the inhibitory effect of CGRP (i.c.v.). It is concluded that centrally administered somatostatin diminished the inhibitory action of CGRP (i.c.v.) on pancreatic secretion, probably via inhibiting autonomic (sympathetic) nerve excitation at the central site. PMID- 9862390 TI - Hyperglycemia suppresses the sympatho-adrenal response to hypoxia, but not to handling stress. AB - We hypothesized that the ability of prior hyperglycemia to suppress the sympatho adrenal response would depend on the type of stress. To test this hypothesis, hyperglycemia was induced in chronically catheterized rats, before submitting them to either hypoxia (7.5% O2) or handling stress. Central venous blood samples were drawn for the determination of plasma glucose, epinephrine (EPI), norepinephrine (NOR) and insulin concentrations. Hypoxia caused significant increases in plasma EPI and NOR concentrations (deltaEPI = + 2.95+/-0.68 nmol/l, deltaNOR = + 12.45+/-1.29 nmol/l). Hyperglycemia, antecedent to hypoxia, dose dependently reduced the sympatho-adrenal response. In contrast, the sympatho adrenal response to handling stress was not affected by even marked antecedent hyperglycemia (deltaEPI = + 2.48+/-0.46 nmol/l, deltaNOR = + 3.12+/-0.69 nmol/l at glucose = 20.7+/-0.6 mmol/l; vs. deltaEPI = + 2.48 + 0.58 nmol/l, deltaNOR= +2.97+/-0.11 nmol/l at glucose = 6.77+/-0.17 mg/dl). Thus, antecedent hyperglycemia suppresses the hypoxia-induced activation of both the sympathetic nerves and the adrenal medulla, but not the activation induced by handling. We conclude that the ability of hyperglycemia to suppress sympathetic activation depends on the stress producing the activation. We therefore speculate that hypoxic stress has a metabolic component to its central activation that handling stress does not. PMID- 9862391 TI - Facilitation of gastric motility induced by portal infusion of hyper- and hypotonic solution in rats. AB - The effects of the portal infusion of hyper- and hypotonic solution on gastric motility in rats were investigated. The infusion of hypertonic saline into the portal vein (portal infusion) elicited a significant enhancement of gastric contractile activity. The portal infusion of water also produced this enhancement. However, the portal infusion of isotonic saline showed no significant enhancement; nor did the infusion of water and hypertonic saline into the jugular vein. Sectioning of the hepatic branch of the vagus nerve (hepatic vagus) eliminated the enhanced responses of the gastric motility. It is therefore concluded that hepatoportal osmoreceptive afferent signals affect the gastric motility by way of the hepatic vagus. These effects on osmolarity revealed that hypotonic stimulation is more effective than hypertonic stimulation for the enhancement of motility. Sectioning of the dorsal subdiaphragmatic vagus, which includes the dorsal gastric and celiac branch, did not eliminate these responses. Sectioning of the ventral gastric vagus, in contrast, did eliminate the responses. These results suggest that vagal preganglionic neurons in the left dorsal motor nucleus of the vagus play a role in enhancement of gastric motility observed in the present research. PMID- 9862392 TI - The effect of distension of the uterus on plasma renin activity (PRA) in anaesthetized pigs. AB - It has recently been shown that distension of the uterus in anaesthetized pigs causes reflex haemodynamic responses through efferent sympathetic mechanisms. The present study was undertaken to determine whether these mechanisms include activation of the renin-angiotensin system. The same methods were used in 14 pigs which were anaesthetized with alpha-chloralose and artificially ventilated. Balloons positioned within the uterus were distended for periods of 30 min by injecting 20 ml of warm Ringer solution. The responses of arterial blood pressure and heart rate were respectively prevented by blockade of alpha-adrenergic receptors with phentolamine and atrial pacing. Changes in plasma renin activity (PRA) were assessed during the last minute of distension by radioimmunoassay of angiotensin I. In each of 10 pigs, distension of the uterus (mean uterine transmural pressure of 17 mmHg) caused an increase in PRA in the absence of changes of interfering haemodynamic variables. In the remaining four pigs, this response was graded by step increments of the distension. The increase in PRA caused by uterine distension was abolished by bilateral section of the renal nerves (five pigs) or by blockade of beta-adrenergic receptors with propranolol. The present study showed that distension of the uterus in anaesthetized pigs primarily caused a reflex increase in PRA. This reflex response was mediated by renal nerves and involved beta-adrenergic receptors. PMID- 9862394 TI - Baroreflex control of muscle sympathetic nerve activity is attenuated in the elderly. AB - To determine whether the baroreflex control of sympathetic nerve activity is attenuated in the elderly, muscle sympathetic nerve activity (MSNA) from the tibial nerve was monitored using microneurography, and heart rate and blood pressure were recorded during the depressor (phase II) or pressor (phase IV) period to Valsalva's maneuver in 10 younger subjects and 7 aged subjects. The baroreflex slope for heart rate showed attenuation in the aged subjects during the pressor phase but not during the depressor phase, the baroreflex slope for MSNA was also attenuated in the aged subjects during the pressor and tended to be attenuated during the depressor phases. These data suggest impaired baroreflex function for both heart rate and sympathetic nerve activity in the elderly. PMID- 9862393 TI - Receptor subtype specific effects of GABA agonists on neurons receiving aortic depressor nerve inputs within the nucleus of the solitary tract. AB - The inhibitory amino acid gamma amino butyrate (GABA) has been shown to profoundly alter the integration of arterial baroreceptor inputs within the nucleus of the solitary tract (NTS). However, the relative roles of the major GABA receptor subtypes, the GABA(A) and the GABA(B) receptors, in the modulation of monosynaptic compared to polysynaptic afferent transmission within the NTS remain uncharacterized. In anesthetized rats, three types of NTS neuron were identified by their responses to aortic depressor nerve (ADN) stimulation; monosynaptic neurons (MSNs), polysynaptic neurons (PSNs) and ADN non-evoked neurons (NENs). Selective GABA(A) and GABA(B) agonists were applied to these neurons using iontophoretic techniques. The endogenous ligand GABA (2 mM), the selective GABA(A) agonist muscimol (0.04 and 0.02 mM) and the GABA(B) agonist baclofen (10 mM) all inhibited the spontaneous discharge of MSNs, PSNs and NENs (P < 0.01 for each group). In addition, GABA, muscimol and baclofen also inhibited ADN evoked discharge in both MSNs and PSNs (P < 0.05 for each group). Both GABA and baclofen significantly inhibited ADN evoked discharge in PSNs to a greater extent than in MSNs (P < 0.05 for each comparison). Muscimol at both doses, however, similarly inhibited ADN evoked discharge in both MSNs and PSNs. Examination of action potential amplitude and co-iontophoretic application of glutamate and GABA agonists suggested that GABA and muscimol induced inhibition were likely to be post-synaptic in origin, while baclofen produced both pre synaptic and post-synaptic inhibition, depending upon the cell. In conclusion, GABA can influence baroreceptor afferent integration through both pre-synaptic and post-synaptic mechanisms. Furthermore, the effects of GABA(B) agonists are variable depending upon the level of afferent integration, with MSNs being generally less sensitive than PSNs. PMID- 9862395 TI - A dysautonomia case of Guillain-Barre syndrome with recovery: monitored by Composite Autonomic Scoring Scale. AB - To investigate the usefulness of the Composite Autonomic Scoring Scale (CASS) as an indication for autonomic dysfunction with Guillain-Barre syndrome, we quantitated autonomic deficits on follow-up using CASS in a patient with Guillain Barre syndrome who did not have any autonomic symptoms and had good clinical recovery. Using the CASS we found the patient to have mild autonomic dysfunction but with a period of recovery. The scale showed that adrenergic deficits improved quickly, sudomotor deficits recovered moderately and cardiovagal deficits did not improve. These results suggested that the patient with Guillain-Barre syndrome might have dysautonomia although she did not have any autonomic symptoms. Results also suggest that there might be difference on the degree of improvement among adrenergic, sudomotor and cardiovagal deficits in Guillain-Barre syndrome. The CASS may be a sensitive tool for the detection of autonomic dysfunction in Guillain-Barre syndrome. PMID- 9862396 TI - Neurobiological similarities in antidepressant sleep deprivation and psychostimulant use: a psychostimulant theory of antidepressant sleep deprivation. AB - This paper attempts to summarize the evidence for the hypothesis that psychostimulant-like neurotransmitter processes within certain regions of the limbic system induce the positive effects of antidepressant sleep deprivation (SD). Preclinical and human studies indicate similar neurobiological effects of psychostimulants such as amphetamines, cocaine and SD. In clinical use, SD and psychostimulants have similar characteristics and behavioral effects. Furthermore, acute psychostimulant challenge decreases limbic metabolism in imaging studies, and SD decreases elevated limbic metabolism in SD responders, indicating that psychostimulant-like neurotransmitter release could decrease limbic metabolism in SD responders. Most antidepressant pharmacotherapies change the reactivity of the dopamine system, and a decrease of presynaptic dopamine or postsynaptic availability can induce depression. Sleep is accompanied by a reduction of catecholamine release and those processes which are increased by psychostimulants. It is concluded that a proposed regional postsynaptic deficit in catecholaminergic neurotransmission can be overcome either acutely by enhanced release during SD or psychostimulant use, or chronically by changes in receptor sensitivity or gene expression due to antidepressant therapies. A postsynaptic deficit in these areas becomes evident if presynaptic release is reduced in conditions such as sleep. Therefore, sleep is depressiogenic for predisposed individuals and the reduction of sleep avoids understimulation of subsensitive postsynaptic processes, which are enhanced by psychostimulants. PMID- 9862397 TI - The cannabinoid CB1 receptor antagonist SR141716A attenuates the memory impairment produced by delta9-tetrahydrocannabinol or anandamide. AB - The administration of delta9-tetrahydrocannabinol (THC), the principle psychoactive ingredient in marijuana, or the endogenous cannabinoid anandamide, has been shown to impair recent memory. The purpose of the present investigation was to determine if the cannabinoid CB1 receptor antagonist SR141716A could attenuate THC- or anandamide-induced memory impairment, and to assess the effects on memory of SR141716A alone. Memory was assessed in rats well-trained in a two component instrumental discrimination task, consisting of a conditional discrimination, and a non-match-to-position to assess recent or working memory. SR141716A (0.0-2.0 mg/kg) had no effect on either the conditional discrimination or the non-match-to-position. However, SR141716A (0.0-2.0 mg/kg) attenuated the memory impairment produced by THC (2.0 or 4.0 mg/kg) as indexed by an enhancement of performance in the non-match-to-position. When administered to rats pretreated with anandamide (2.0 mg/kg), SR141716A (0.0-2.5 mg/kg) impaired performance in the conditional discrimination at the highest dose. This was interpreted as a deficit in some capacity unrelated to memory (e.g., motor impairment). However, lower doses of SR141716A (0.1 and 0.5 mg/kg) attenuated the anandamide-induced impairment of performance in the non-match-to-position without affecting the conditional discrimination. This is the first report that the memory impairment produced by anandamide can be attenuated by a cannabinoid antagonist; results suggest that anandamide-induced memory disruption is mediated by CB receptors. PMID- 9862398 TI - The mu opioid irreversible antagonist beta-funaltrexamine differentiates the discriminative stimulus effects of opioids with high and low efficacy at the mu opioid receptor. AB - The purpose of the present study was to determine the relative intrinsic efficacy of various opioids using the irreversible mu opioid antagonist beta funaltrexamine (betaFNA). To this end, pigeons were trained to discriminate 3.0 (n=6) or 1.8 (n=1) mg/kg morphine from distilled water in a two-key, food reinforced, drug discrimination procedure. The mu opioids fentanyl, l-methadone, buprenorphine, butorphanol, nalorphine, nalbuphine and levallorphan, as well as the delta opioid BW373U86, substituted completely for the morphine stimulus. The stimulus effects of morphine were antagonized (i.e., produced a significant increase in the ED50 value) by a 10 mg/kg but not a 5 mg/kg dose of betaFNA. Antagonist effects of betaFNA were observed following a 2-h pretreatment, but not following 26-, 50-, 74-, 98- or 146-h pretreatments. The stimulus effects produced by fentanyl, l-methadone and buprenorphine were not antagonized by doses of betaFNA as high as 20, 10 and 10 mg/kg, respectively. The lowest dose of betaFNA required to antagonize the stimulus effects of butorphanol was 10 mg/kg, whereas the effects of nalorphine, nalbuphine and levallorphan were antagonized by a dose of betaFNA as low as 5 mg/kg. The delta BW373U86 substituted for the morphine stimulus, and this effect was not antagonized by 10 mg/kg betaFNA. The pkB values for naloxone (1.0 mg/kg) against the stimulus effects of fentanyl (6.70) and morphine (6.52) were considerably higher than that for BW373U86 (4.60), indicating further that the morphine-like stimulus effects produced by BW373U86 were not mediated by activity at the mu opioid receptor. These findings indicate that the strategy of irreversible antagonism can be used effectively to differentiate opioids with varying degrees of intrinsic efficacy at the mu opioid receptor in a pigeon drug discrimination procedure. In particular, the ranking of these drugs by relative intrinsic efficacy at the mu opioid receptor is: l methadone=fentanyl> or =buprenorphine> or =morphine> or =butorphanol>nalorphine=nalbuphine=levallorphan. Additionally, the short-acting effect of betaFNA in the pigeon suggests that the recovery of mu opioid receptor function varies across species. PMID- 9862399 TI - Buprenorphine alters ethanol self-administration in rats: dose-response and time dependent effects. AB - Buprenorphine is a partial opioid agonist derived from thebaine and has high affinity for mu and kappa opioid receptors. The present study investigated dose response (0.03, 0.15, 0.3, 3 mg/kg) and time-dependent effects of buprenorphine (1.5 or 4 h post-treatment) on EtOH self-administration in outbred Sprague-Dawley rats. Freely feeding and drinking rats were trained to initiate EtOH self administration for 1 h daily using the ascending concentration procedure, wherein they were provided with increasing concentrations of EtOH at 2, 5, 7, 9 and 11% (v/v), respectively. Water was concurrently available with each concentration. Animals were maintained on a given concentration of EtOH for 5 days. By day 21, animals began their stabilization on the 11% regimen and remained on this concentration throughout the remainder of the study. EtOH and water consumption were recorded daily at both 10- and 60-min intervals. At 1.5 h post buprenorphine, all test doses greatly suppressed both EtOH and water intake at the 10-min interval. At the 60-min interval, all but the lowest dose (0.03 mg/kg) significantly suppressed EtOH intake, while only the highest dose (3 mg/kg) suppressed water intake. In contrast to the suppressant profile observed at 1.5 h post-buprenorphine, at 4 h post-buprenorphine the lower doses (0.03 and 0.15 mg/kg) significantly increased EtOH intake while the higher doses (0.3 and 3 mg/kg) continued to suppress intake. None of the doses of buprenorphine altered water intake 4 h post-buprenorphine. The results support previous research demonstrating the utility of low doses of buprenorphine in suppressing behavior rewarded by a non-opioid drug. PMID- 9862400 TI - Influence of nicotine on simulator flight performance in non-smokers. AB - In a placebo-controlled study, we investigated the influence of nicotine on late day aviation performance in 15 non-smoking subjects. In a within-subjects design, subjects were tested on 2 days, each lasting 8 h and consisting of three 75-min simulator flights (late-afternoon practice, evening test, night test). Prior to each test, subjects received either nicotine polacrilex 2 mg or placebo gum. As expected, overall performance was significantly better after nicotine, compared to placebo (P < 0.01). Post-hoc analysis of individual flight tasks showed that nicotine improved scores on approach to landing, a task which appears to require sustained attention. We conclude that nicotine may improve late-day flight performance in non-smoking aviators. PMID- 9862401 TI - Sensitivity to cocaine and amphetamine among mice selectively bred for differential cocaine sensitivity. AB - Selective breeding of mice for differences in response to a drug offers a powerful means for testing hypotheses regarding underlying mechanisms and relationships between drug-induced behaviors. Starting from a heterogeneous stock of mice, we have selectively bred lines of mice for extreme differences in their locomotor response to 10 mg/kg cocaine HCl. Selection pressure has been maintained for 12 generations and has resulted in two cocaine sensitive (CAHI) and two cocaine insensitive (CALO) lines. Across the generations of selection, the CAHI lines showed progressively greater amounts of cocaine-induced locomotion, with mice from the S12 generation traveling over 21,000 cm/30 min. following 10 mg/kg cocaine. The CALO lines, in contrast, did not substantially diverge from control values until the S8 generation. By generation 12, however, the LO lines traveled no further following 10 mg/kg cocaine (7000 cm/30 min), than they did following an initial saline injection. Cocaine and amphetamine dose response analyses were conducted on drug-naive mice from the tenth generation. The CAHI lines were extremely sensitive to the locomotor activating effects of all doses of cocaine, displaying from 2- to 6-fold greater amounts of cocaine induced locomotion than the CALO lines. The CALO lines, in contrast, were completely insensitive to the psychomotor stimulant effects of cocaine. The CAHI lines were also more sensitive to the locomotor activating effects of amphetamine. Both lines showed dose-dependent amphetamine-induced locomotion that peaked at 3 mg/kg. However, at all doses, the CAHI lines showed a 2- to 4-fold greater amount of locomotion than CALO lines. Thus, the sensitivity to cocaine developed through selection using a single dose of cocaine has generalized to a range of doses of cocaine and to at least one other psychostimulant. PMID- 9862402 TI - Repeated administration of ephedrine induces behavioral sensitization in rats. AB - Systemic injection of the sympathomimetic agent ephedrine (EPH) stimulates locomotion in drug-naive rats, an effect that may be dependent on the enantiomer of EPH employed [(-)-EPH or (+)-EPH]. The present experiments examined the effects of repeated EPH exposure on locomotion in rats to assess whether these treatments result in drug tolerance or sensitization. In experiment 1, adult male rats were injected once daily with 0, 10, 20, or 40 mg/kg (-)-EPH (IP) on each of 11 days. Locomotor activity was assessed for 60 min after drug injection. Acute exposure to (-)-EPH treatment increased locomotion for animals receiving 20 or 40 mg/kg, and this effect was augmented after 11 days of drug administration. A vehicle-only injection was given to all animals on day 12 to determine the influence of environmental cues on sensitization. On day 13, all rats were injected with 10 mg/kg cocaine HCl to assess whether repeated (-)-EPH exposure produced a cross-sensitization to cocaine (10 mg/kg, IP). Only rats treated repeatedly with 40 mg/kg (-)-EPH exhibited increases in cocaine-stimulated locomotion relative to saline-treated rats. In experiment 2, repeated exposure to (+)-EPH, 40 mg/kg, but not 20 mg/kg, increased activity and demonstrated the development of sensitization. Cross-sensitization to cocaine (10 mg/kg, IP) was not evident following treatment with either concentration of (+)-EPH. There was no evidence that contextual events alone played a role in the effects observed here. PMID- 9862403 TI - Tolerance and cross-tolerance to the rate-suppressing effects of opioids in butorphanol-treated rats: influence of maintenance dose and relative efficacy at the mu receptor. AB - The purpose of the present investigation was to examine the development of tolerance to the rate-suppressing effects of mu and kappa opioids in rats administered either 3.0 (low) or 30 (high) mg/kg per day of butorphanol, an opioid with low relative efficacy at the mu receptor. The mu opioids butorphanol, buprenorphine, morphine, fentanyl and sufentanil, and the kappa opioid U50,488 dose-dependently suppressed responding under all conditions examined. In rats administered the low maintenance dose of butorphanol, tolerance developed to the effects of butorphanol, buprenorphine and morphine, but not to fentanyl and sufentanil. In rats administered the high maintenance dose, tolerance developed to all of the mu opioids examined. In both treatment groups, the degree to which tolerance developed was greater for butorphanol and buprenorphine than for morphine, fentanyl and sufentanil; and the degree to which tolerance developed to these mu opioids was greater in rats administered the high maintenance dose of butorphanol. The tolerance that developed to morphine, fentanyl and sufentanil was not altered when tested at both 23 and 47 h following the previous maintenance dose of butorphanol, suggesting that these changes were not due to any acute pharmacological interactions between butorphanol and the test compound (i.e., antagonism). Tolerance was also conferred to the kappa opioid U50,488 in both groups of rats, and in rats administered the high maintenance dose, this effect was obtained when tested 23 and 47 h following the previous maintenance dose of butorphanol. Physical dependence developed in rats administered the high maintenance dose of butorphanol, as evidenced by the development of enhanced sensitivity to the rate-suppressing effects of naloxone, and the finding that 30 mg/kg naloxone decreased body weight in a time-dependent manner. No physical dependence was apparent in rats administered the low maintenance dose of butorphanol. These data suggest that during chronic treatment with butorphanol, (1) greater degrees of tolerance are conferred to drugs possessing low efficacy at the mu opioid receptor, (2) tolerance is enhanced as the maintenance dose of the toleragen is increased, and (3) mu-opioid tolerance may be observed under conditions that do not produce mu-opioid dependence. PMID- 9862404 TI - Evidence for a functional interaction between 5-HT1A and 5-HT2 receptors in rats. AB - Evidence of a functional interaction between serotonin 5-HT1A and 5-HT2 receptor subtypes has been compromised by incomplete experimental designs and conflicting data. To test for such an interaction, combinations of the 5-HT1A agonist 8-OH DPAT and the 5-HT2 agonist DOI were administered to rats prior to testing of locomotor activity in the Behavioral Pattern Monitor (BPM). The BPM is an activity and holeboard chamber that enables analyses of quantitative and qualitative changes in locomotor and investigatory activity. Dose-response studies of 8-OH-DPAT and DOI alone and in the presence of selected doses of the other drug were performed in order to allow isobolographic analysis, which characterizes the relationship of two drugs as either additive (no interaction), supra-additive, or infra-additive. Rats treated with saline, 8-OH-DPAT (6.25, 12.5, 25, or 50 microg/kg SC), DOI (0.15, 0.3, or 0.6 mg/kg SC), or selected combinations of both drugs were tested in the BPM for 1 h. Isobolographic analysis of the effects on locomotor activity revealed that 8-OH-DPAT and DOI interact in an infra-additive manner. Thus, at a functional level, 5-HT1A and 5 HT2 receptors interact antagonistically in the modulation of locomotor activity. PMID- 9862405 TI - Seroquel, clozapine and chlorpromazine restore sensorimotor gating in ketamine treated rats. AB - Sensorimotor gating of the startle reflex measured by prepulse inhibition (PPI) is impaired in schizophrenia patients and in rats treated with either dopamine (DA) agonists or with N-methyl-D-aspartate (NMDA) antagonists. While both typical and atypical antipsychotics restore PPI in DA agonist-treated rats, studies thus far have demonstrated that only atypical antipsychotics restore PPI in rats treated with NMDA antagonists. This model for predicting atypical antipsychotic properties has been studied extensively in rats, and there is interest in moving these studies into humans, where the NMDA antagonist ketamine is also reported to significantly reduce PPI. In anticipation of such studies, and to facilitate the use of this model in humans, we examined the effects of high and low potency typical antipsychotics (haloperidol and chlorpromazine), the atypical antipsychotic clozapine, and the putative atypical antipsychotic, Seroquel, on ketamine-disrupted PPI in rats, across a range of ketamine that produced submaximal, as well as maximal disruptions of PPI. Ketamine dose-dependently reduced PPI, and this effect was significantly opposed by Seroquel, clozapine and chlorpromazine, but not haloperidol. The effects of chlorpromazine on ketamine disrupted PPI demonstrate that the ability of antipsychotics to restore PPI in NMDA antagonist-treated rats is not specific to clinically atypical antipsychotics. Receptor properties shared by Seroquel, clozapine and chlorpromazine, but not haloperidol, may implicate critical substrates in the NMDA antagonist-induced disruption of PPI. PMID- 9862406 TI - Haloperidol, raclopride, and eticlopride induce microcatalepsy during operant performance in rats, but clozapine and SCH 23390 do not. AB - The purpose of this work was (1) to assess the ability of selected antipsychotic and comparison drugs to induce arrest of movement phenomena during operant responding and (2) to evaluate the capacity of muscarinic anitcholinergics to block such effects. The effects of haloperidol (0.02-0.12 mg/kg, i.p., 45 min), raclopride (0.05-0.80 mg/kg, i.p., 30 min) eticlopride (0.02-0.16 mg/kg, i.p., 45 min), clozapine (1.0-8.0 mg/kg, i.p., 60 min) and SCH 23390 (0.01-0.16 mg/kg, i.p., 30 min) were administered to rats for 4 weeks in a between-groups dosing design. Operant responses in 15 min and the maximum duration of the rat's muzzle entry into the reinforcement dipper well (the measure of arrest of movement that reflected microcatalepsy) were the quantitative measures of behavior. The D2 antagonists dose-relatedly decreased operant responding and increased maximum muzzle duration, effects that were significantly reversed by the anticholinergic scopolamine (0.1 mg/kg) or atropine (6.0 mg/kg). Although the atypical antipsychotic drug clozapine and the selective D1 antagonist SCH 23390 both significantly reduced operant responding, these drugs did not produce microcatalepsy. The results suggested that microcatalepsy expressed in the context of ongoing operant behavior may model low-dose extrapyramidal side effects. PMID- 9862407 TI - Comparison of the discriminative and antinociceptive effects of morphine and its glucuronide metabolites after central or systemic administration in the rat. AB - The potential of centrally (ICV) or systemically (SC) administered M6G to substitute for morphine in a drug discrimination task was characterized in the present study. Rats with a cannula in the lateral cerebral ventricle were trained to discriminate between injections of morphine (3 mg/kg, SC) and saline using a discrete-trial avoidance/escape procedure. Substitution tests were conducted with SC or ICV morphine, morphine-3-beta-D-glucuronide (M3G), or morphine-6-beta-D glucuronide (M6G) and response latency in a tail-flick test was measured before each session began. The stimulus effects of morphine (ED50=1.02 mg/kg SC or 2.1 microg/kg ICV) were fully shared by M6G, with potency dependent on route of administration (ED50=3.12 mg/kg SC or 0.34 microg/kg ICV). The stimulus effects of M6G were highly correlated with its antinociceptive activity (r=0.84 SC or 0.46 ICV) and, at equipotent systemic doses, they lasted longer (t1/2=391 min) than those of morphine (t1/2=185 min). M3G was inactive in both procedures by both routes of administration. Naltrexone SC, given 30 min prior to testing, completely attenuated the stimulus effects of ICV M6G (AD50=0.011 mg/kg), indicating that they are mediated by opioid receptors. The results of this study suggest that M6G might contribute to the interoceptive effects of morphine that underlie its potential for abuse. PMID- 9862408 TI - Temporal effects of nicotine nasal spray and gum on nicotine withdrawal symptoms. AB - Nicotine nasal spray and nicotine gum have been found to be effective in relieving nicotine withdrawal symptoms. In this randomized single-blind study, 91 cigarette smokers were randomly assigned to a single 1 mg dose of active nicotine nasal spray (n=29), active 4 mg nicotine gum (n=31), saline placebo nasal spray (n=16) or placebo gum (n=15). Following overnight abstinence, subjects repeatedly completed visual analog scales for assessing nicotine withdrawal symptoms over 30 min preceding (time -30 min to time 0) and 120 min following a single dose of study medication. This sequence was performed 3 times during the day. Nicotine withdrawal symptoms were assessed on a 41-point visual analog scale (1=no withdrawal, 41=extreme withdrawal). At the initial session only, blood samples for serum nicotine levels were taken at baseline, then at 5, 10, 30 and 120 min following study drug administration. The mean (+/-SD) age of the subjects was 38.6 (+/-10.1) years, 48% were females, smoking rate was 24.5 (+/-7.8) cigarettes per day, and years of smoking was 19.9 (+/-10.0). A single 1 mg dose of nicotine nasal spray provided more immediate relief for craving for a cigarette compared to a single 4 mg dose of nicotine gum. Serum venous nicotine levels for the active nicotine nasal spray and nicotine gum were comparable at 5 and 10 min while the levels were higher for nicotine gum at 30 and 120 min. Changes in withdrawal symptoms were not found to be related to serum venous nicotine levels. Our findings provide a rationale for the as needed use of nicotine nasal spray to control withdrawal symptoms, possibly in combination with other medications with longer acting effects. PMID- 9862409 TI - Evidence that GABA transmission mediates context-specific extinction of learned fear. AB - Six experiments used rats to study the effects of the beta-carboline FG 7142 on extinction of fear responses (freezing) to an auditory cue that had signalled footshock. Subcutaneous injection of FG 7142 interfered with the development of extinction without having any detectable effect on the rats' levels of fear prior to extinction. Injection of FG 7142 also reversed extinction, partially reinstating fear responses that had been extinguished previously. A similar reinstatement of extinguished fear was seen when rats were tested for fear of the cue in a different chamber. The reinstatement produced by FG 7142 and that caused by context shift were not additive: FG 7142 did not increase extinguished fear if rats were tested in the different chamber. Finally, FG 7142 had no detectable effect on the latent inhibition of fear produced by repeatedly presenting the cue alone before conditioning with shock, even though this inhibition, like extinction, was affected by a shift in context. The present findings indicate that GABA transmission at GABA(A) receptors is involved in the inhibition of extinguished fear, and that this effect of GABA is regulated by those cues that constitute the extinction context. PMID- 9862410 TI - The effects of a low dose of caffeine on cognitive performance. AB - There is little evidence concerning the effects of caffeine in doses typical of one cup of tea. The present study investigated the effect of 60 mg caffeine, consumed in either tea or hot water, on performance on a subset of the CANTAB test battery. Eight males participated in a practice session and four test sessions. In each test session, the participant consumed a different hot beverage and then, over approximately 90 min, completed nine tests from the CANTAB battery. The four beverages were created by crossing beverage identity (tea or hot water) and caffeine dose (0 or 60 mg). Significant speeding of reaction time by caffeine consumption was found in pattern recognition, delayed match to sample, and match to sample visual search. The effect on reaction time of 60 mg caffeine can be detected, and may be evident within minutes of consumption. Objective reports of immediate beneficial effects of consumption, most research has postponed measurement to coincide with peak plasma caffeine levels (Blanchard and Sawers 1983). The intention of the present study was to investigate the effects of consuming a single cup of tea on a variety of cognitive tests. Testing began immediately after consumption and lasted approximately 80 min. PMID- 9862411 TI - Clomipramine enhances the cortisol response to 5-HTP: implications for the therapeutic role of 5-HT2 receptors. AB - We measured the cortisol response to the 5-HT precursor, 5-hydroxytryptophan, (5 HTP) in seven patients with major depression before and after 8 weeks treatment with the tricyclic antidepressant, clomipramine. The cortisol response to 5-HTP was significantly increased following clomipramine treatment, suggesting that clomipramine, like selective serotonin re-uptake inhibitors (SSRIs), enhances this 5-HT2 receptor mediated response. Because other tricyclic antidepressants do not increase 5-HTP-mediated cortisol release, it seems unlikely that enhancement of 5-HT2 receptor function is a critical mechanism for antidepressant action. However, facilitation of neurotransmission at 5-HT2 receptors could account for the efficacy of clomipramine and SSRIs in the treatment of obsessive compulsive disorder and also for their liability to cause orgasmic dysfunction. PMID- 9862412 TI - Oxygen and cognitive performance: the temporal relationship between hyperoxia and enhanced memory. AB - Oxygen administration coinciding with word presentation enhances word recall in humans, suggesting that elevated levels of circulating blood oxygen may be available to neural memory consolidation processes. This double-blind experiment examined the relationship between blood oxygen levels and cognitive performance when oxygen was inspired for 2 min at different times relative to a simple word recall task, forward digit span and backward digit span. Transient hyperoxia, measured by haemoglobin-bound oxygen, was evident following oxygen inspiration. Neither forward nor backward digit span was affected by oxygen administration. Word recall (12 min following word presentation) was enhanced when oxygen was administered 5 min prior to, immediately before or immediately following word presentation; but not 10 min prior to, 5 min following nor 10 min following, word presentation. These data suggest that oxygen administration can selectively enhance aspects of cognitive performance and support a hypothesis whereby supplemental blood oxygen is sequestered by neural mechanisms involved in memory consolidation. PMID- 9862413 TI - The involvement of poly(ADP-ribose) polymerase in the oxidative stress responses in plants. AB - In plants many biotic and abiotic stresses can cause secondary oxidative stress. Earlier work showed that, depending on the severity of the oxidative stress, plants can activate either cell protective genes or programmed cell death (PCD). Poly(ADP-ribose) polymerase (PARP) has been implicated as one of the enzymes in the apoptotic pathways induced by DNA damaging agents or oxidative stress. We show that in cultured soybean cells, PARP is involved in responses to mild and severe oxidative stresses, by mediating DNA repair and PCD processes, respectively. Addition of PARP inhibitors reduced the degree of cell death triggered by H2O2. Two windows of NAD consumption after H2O2 treatment were detected. Experiments with transient overexpression of Arabidopsis PARP cDNA promoted DNA repair and inhibited cell death caused by mild oxidative stress. However, following severe stress PARP overexpression increased cell death. Expression of antisense PARP produced the opposite effects: an increase in DNA nicks and inhibition of cell death at high, but not mild doses of H2O2. PMID- 9862414 TI - Selective inhibition of NF-kappaB activation by the flavonoid hepatoprotector silymarin in HepG2. Evidence for different activating pathways. AB - The bioflavonoid silymarin is found to potently suppress both nuclear factor kappa-B (NF-kappaB)-DNA binding activity and its dependent gene expression induced by okadaic acid in the hepatoma cell line HepG2. Surprisingly, tumor necrosis factor-alpha-induced NF-kappaB activation was not affected by silymarin, thus demonstrating a pathway-dependent inhibition by silymarin. Many genes encoding the proteins of the hepatic acute phase response are under the control of the transcription factor NF-kappaB, a key regulator in the inflammatory and immune reactions. Thus, the inhibitory effect of silymarin on NF-kappaB activation could be involved in its hepatoprotective property. PMID- 9862415 TI - Apoptosis induced by hydrogen peroxide is mediated by decreased superoxide anion concentration and reduction of intracellular milieu. AB - Hydrogen peroxide (H2O2) is considered to be a mediator of apoptotic cell death but the mechanism by which it induces apoptosis is unclear. Here, we show that cells undergoing apoptosis from exposure to H2O2 display a significant decrease in intracellular concentration of superoxide (O2-) which is associated with a reduction of the intracellular milieu, as measured by an increase in the GSH/GSSG ratio and a decrease in intracellular pH. The notion that a decrease in intracellular O2- concentration triggers apoptosis is supported by the observation that H2O2-mediated apoptosis could be retarded in cells in which the intracellular O2- concentration is maintained at or above the cellular baseline level by inhibition of the major O2- scavenger superoxide dismutase (Cu/Zn SOD). Taken together, our observations indicate that a decrease in the intracellular O2 concentration, reduction and acidification of the intracellular milieu constitute a signal for H2O2-mediated apoptosis, thereby inducing a reductive as opposed to an oxidative stress. PMID- 9862416 TI - Hypericin-induced photosensitization of HeLa cells leads to apoptosis or necrosis. Involvement of cytochrome c and procaspase-3 activation in the mechanism of apoptosis. AB - Here we report that photoactivated hypericin can induce either apoptosis or necrosis in HeLa cells. Under apoptotic conditions the cleavage of poly(ADP ribose) polymerase (PARP) into the 85-kDa product is blocked by the caspase inhibitors benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk) and benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethylketone (z-DEVD-fmk). Both inhibitors protect cells from apoptosis but cannot prevent hypericin-induced necrosis. Conversely, HeLa cells overexpressing the viral cytokine response modifier A (CrmA), which inhibits caspase-1 and -8, still undergo hypericin induced apoptosis and necrosis. Evidence is provided for the release of mitochondrial cytochrome c in the cytosol and for procaspase-3 activation in the hypericin-induced cell killing. PMID- 9862417 TI - Down-regulation of protein kinase C alpha and gamma and enhanced TPA-induced neurite formation in DAN-transfected neuroblastoma cells. AB - DAN gene was first isolated by differential screening between rat 3Y1 and v-src transformed 3Y1 cells and showed a tumor-suppressive activity toward v-src transformed 3Y1 cells. When DAN-transfected neuroblastoma cells were treated with a tumor promoter phorbol ester, TPA, neurite-like processes appeared within 2 h whereas no apparent change was observed in the parent and vector-transfected cells up to 8 h. This suggests some difference in TPA-receptor, protein kinase C (PKC), between DAN-transfectants and the control cells. DAN-transfected SH-SY5Y cells showed complete loss in PKCalpha and a large decrease in PKCgamma. Similar down-regulation in PKCalpha and PKCgamma was also observed in DAN-transfected Ha ras-transformed NIH 3T3 cells. The decreased level of PKCalpha was partially recovered after treatment with a calpain inhibitor, ZLLH. A 150-kDa proteolytic product of a calpain-specific substrate, non-erythroid alpha-spectrin, was detectable in DAN-transfected SH-SY5Y cells but not in the parent or vector transfected control cells. This suggests that DAN-transfected cells contain activated calpain which may cause down-regulation of PKC and hence induce the altered TPA response. PMID- 9862418 TI - Expression of lectin-like oxidized low density lipoprotein receptor-1 in human and murine macrophages: upregulated expression by TNF-alpha. AB - Uptake of oxidized low density lipoprotein (Ox-LDL) and subsequent foam cell transformation have been implicated in early atherogenesis. Although multiple molecules, including class A and B scavenger receptors, have been identified as Ox-LDL receptors, additional receptors may also be involved in this process. Here, we provide evidence that lectin-like Ox-LDL receptor-1 (LOX-1), a novel Ox LDL receptor initially identified in vascular endothelial cells, is also expressed in macrophages in humans and mice. Expression of LOX-1 can be induced after macrophage-like differentiation in vitro in human peripheral blood monocytes and the related cell line THP-1 cells. Furthermore, LOX-1 expression can also be detected in resident peritoneal macrophages, and can be upregulated by an inflammatory cytokine TNF-alpha. These results suggest that LOX-1 in macrophages may play an important role in Ox-LDL uptake and subsequent foam cell formation in this cell type. PMID- 9862419 TI - Characterization of the 5'-flanking promoter region of the rat somatostatin receptor subtype 3 gene. AB - We investigated the 5'-flanking promoter region of the rat somatostatin receptor subtype 3 (rSSTR3). Using a cDNA probe, genomic clones containing the 5'-flanking promoter region of the rSSTR3 gene were isolated. A sequence of 5.4 kb directly upstream from the start codon was analyzed and two introns were found in the 5' untranslated region (UTR) of the cDNA sequence. The transcriptional initiation site was determined by 5' rapid amplification of cDNA ends (RACE), primer extension and RNase protection analysis with cerebellar RNA. Two major transcriptional initiation sites were found at position 1040 (tsp1) and -856 (tsp2) relative to the translational initiation site. Like a number of other promoters of G-protein-coupled receptors, the rSSTR3 gene lacks TATA and CAAT motifs and includes G+C-rich regions. Functional analysis of the promoter region by transfecting rSSTR3 luciferase-reporter gene constructs into rat pituitary GH3 cells and HEK 293 cells indicated that a 107-bp region upstream of tsp2 was sufficient to drive transcription. Furthermore a 562-bp region at position -1304 to -1865 upstream of the ATG start codon exerted a negative regulatory effect on transcriptional activity. PMID- 9862420 TI - Chromatin proteins surrounding the d(G-T)n repeats and polyamine influence as revealed by photoaffinity labeling with reactive pd(A-C)6 derivatives. AB - The complementary-addressed modification of DNA and proteins in chromatin using photoreactive derivatives of pd(AC)6 has been studied. These oligonucleotides form complementary complexes with specific DNA sequences and modify both DNA and proteins in the vicinity of these regions, and can be used for investigation of the protein environment in DNA. We have demonstrated that photoreactive derivatives of oligonucleotides can quickly and efficiently modify chromatin proteins and seem to be promising for investigation of perturbations in chromatin structure during the cell cycle. A comparison between modified chromatin from synchronized cells has demonstrated differences in the sets of proteins modified in the S and G1/S phases of the cell cycle. An increase in spermine and spermidine concentrations leads to an increase in modification of definite chromatin proteins. It can be supposed that the B-Z transition that can be stabilized by the presence of natural polyamines is one of the reasons for the presence of single-stranded DNA regions, containing sets of (dG-dT)n and accessible for interaction with complementary oligonucleotides. PMID- 9862421 TI - TNF inhibits insulin induced STAT5 activation in differentiated mouse muscle cells pmi28. AB - Tumor necrosis factor (TNF) plays a central role in the state of insulin resistance leading to type II diabetes. We here describe the crosstalk of TNF with insulin signaling cascades in the mouse muscle cell line pmi28. TNF downregulated insulin induced insulin receptor kinase activity and insulin induced activation of the transcription factor STAT5. Our results provide evidence that the inhibitory crosstalk between TNF and insulin in skeletal muscle cells comprises an interference with the expression of STAT5 regulated genes which may play an important role in the manifestation and/or progression of insulin resistance in muscle cells. PMID- 9862422 TI - Effect of MI-D, a new mesoionic compound, on energy-linked functions of rat liver mitochondria. AB - MI-D (4-phenyl-5-(4-nitro-cinnamoyl)-1,3,4-thiadiazolium-2-phenylami ne chloride), a new mesoionic compound, depressed the phosphorylation efficiency of liver mitochondria as deduced from an accentuated decrease of the respiratory control coefficient and ADP/O ratio. Analysis of segments of the respiratory chain suggested that the MI-D inhibition site is further on than complex I and between complexes II and III. The transmembrane electrical potential (delta psi) was collapsed dependent on MI-D concentration. ATPase activity was dramatically increased by MI-D in intact mitochondria, but inhibited in carbonylcyanide p trifluoromethoxyphenylhydrazone (FCCP)-uncoupled mitochondria. These results suggest that MI-D acts as an uncoupler agent, a property closely related to its structural characteristics. PMID- 9862423 TI - Induction of outward current by orexin-B in mouse peritoneal macrophages. AB - To define effects of novel feeding regulating peptides, orexins, in immunocompetent cells, ion channel activity in mouse peritoneal macrophages was analyzed by the perforated patch-clamp method. Orexin-B (OX-B) induced an outward current at smaller holding potentials than K+ equilibrium potentials. Reversal potentials of OX-B induced current were dependent on external K+ concentrations but not on external Cl- concentration. Orexin-A is less effective than OX-B. Quinine blocked the outward current and tetraethylammonium partially suppressed the current. These results suggest that OX-B can modulate macrophage functions through the activation of Ca2+-dependent K2+ channels. PMID- 9862424 TI - Glyoxylate cycle enzymes are present in liver peroxisomes of alloxan-treated rats. AB - Key enzymes of the glyoxylate cycle, isocitrate lyase (ICL) and malate synthase (MS), have been detected in the liver of alloxan-treated rats. The activity of ICL in rat liver was 0.040 micromol/min/mg protein and the activity of MS was 0.022 micromol/min/mg protein. These enzymes were associated with the peroxisomal fraction. The activities of citrate synthase, malate synthase and malate dehydrogenase detected in the peroxisomal fraction were also increased by alloxan treatment. Isocitrate lyase was partially purified and displayed catalytic and regulatory properties similar to those of the enzyme isolated from the liver of starved rats (Popov, V.N. et al. (1996) FEBS Lett. 391, 87-90). PMID- 9862425 TI - Demonstration of thermal dissipation of absorbed quanta during energy-dependent quenching of chlorophyll fluorescence in photosynthetic membranes. AB - When plant leaves or chloroplasts are exposed to illumination that exceeds their photosynthetic capacity, photoprotective mechanisms such as described by the energy-dependent (non-photochemical) quenching of chlorophyll fluorescence are involved. The protective action is attributed to an increased rate constant for thermal dissipation of absorbed quanta. We applied photoacoustic spectroscopy to monitor thermal dissipation in spinach thylakoid membranes together with simultaneous measurement of chlorophyll fluorescence in the presence of inhibitors of opposite action on the formation of delta pH across the thylakoid membrane (tentoxin and nigericin/valinomycin). A linear relationship between the appearance of fluorescence quenching during formation of the delta pH and the reciprocal variation of thermal dissipation was demonstrated. Dicyclohexylcarbodiimide, which is known to prevent protonation of the minor light-harvesting complexes of photosystem II, significantly reduced the formation of fluorescence quenching and the concurrent increase in thermal dissipation. However, the addition of exogenous ascorbate to activate the xanthophyll de epoxidase increased non-photochemical fluorescence quenching without affecting the measured thermal dissipation. It is concluded that a portion of energy dependent fluorescence quenching that is independent of de-epoxidase activity can be readily measured by photoacoustic spectroscopy as an increase in thermal deactivation processes. PMID- 9862426 TI - Product activation of human erythrocyte AMP deaminase. AB - IMP was found to activate AMP deaminase in crude glucose-depleted human erythrocyte lysates. Activation of the enzyme by IMP is due to prevention of the inhibitory effect of inorganic phosphate. At 1 mM AMP and 2-3 mM phosphate the addition of 2-5 mM IMP accelerates the AMP deamination two to three times. PMID- 9862427 TI - Effects of the mutations Ala30 to Pro and Ala53 to Thr on the physical and morphological properties of alpha-synuclein protein implicated in Parkinson's disease. AB - Alpha-synuclein (alpha-syn) protein has been found in association with the pathological lesions of a number of neurodegenerative diseases. Recently, mutations in the alpha-syn gene have been reported in families susceptible to an inherited form of Parkinson's disease. We report here that human wild-type alpha syn, PD-linked mutant alpha-syn(Ala30Pro) and mutant alpha-syn(Ala53Thr) proteins can self-aggregate and form amyloid-like filaments. The mutant alpha-syn forms more beta-sheet and mature filaments than the wild-type protein. These findings suggest that accumulation of alpha-syn as insoluble deposits of amyloid may play a major role in the pathogenesis of these neurodegenerative diseases. PMID- 9862428 TI - Aggregates from mutant and wild-type alpha-synuclein proteins and NAC peptide induce apoptotic cell death in human neuroblastoma cells by formation of beta sheet and amyloid-like filaments. AB - Alpha-synuclein (alpha-syn) protein and a fragment of it, called NAC, have been found in association with the pathological lesions of a number of neurodegenerative diseases. Recently, mutations in the alpha-syn gene have been reported in families susceptible to an inherited form of Parkinson's disease. We have shown that human wild-type alpha-syn, mutant alpha-syn(Ala30Pro) and mutant alpha-syn(Ala53Thr) proteins can self-aggregate and form amyloid-like filaments. Here we report that aggregates of NAC and alpha-syn proteins induced apoptotic cell death in human neuroblastoma SH-SY5Y cells. These findings indicate that accumulation of alpha-syn and its degradation products may play a major role in the development of the pathogenesis of these neurodegenerative diseases. PMID- 9862429 TI - Prostaglandins prevent inducible nitric oxide synthase protein expression by inhibiting nuclear factor-kappaB activation in J774 macrophages. AB - We investigated the effect of PGE2 and iloprost (a prostacyclin analogue) on inducible nitric oxide synthase (iNOS) protein expression and nuclear factor kappaB (NF-kappaB) activation in lipopolysaccharide (LPS)-stimulated J774 macrophages. Incubation of J774 cells with LPS (10 microg/ml) caused an increase of iNOS protein expression which was prevented in a concentration-dependent fashion by PGE2 (0.1, 1, 10 microM) and iloprost (0.01, 0.1, 1 microM). Electrophoretic mobility shift assay indicated that both prostanoids blocked the activation of NF-kappaB, a transcription factor necessary for NO synthase induction. PGE2 and iloprost also blocked disappearance of I kappaB-alpha from cytosolic fraction and nuclear translocation of NF-kappaB subunits p50 and p65. These results show for the first time that PGE2 and iloprost down-regulate iNOS protein expression by inhibiting NF-kappaB activation and suggest a negative feed back mechanism that may be important for limiting excessive or prolonged NO production in pathological events. PMID- 9862430 TI - Cooperativity and flexibility of active sites in homodimeric transketolase. AB - Here we summarize evidence for non-equivalence of two structurally similar active sites in transketolase and other thiamine-dependent enzymes. This non-equivalence takes place when the enzymes interact with various ligands (inhibitors, cations, coenzyme and substrates). Data on different strains in the structure of the holotransketolase subunits are also given. The above results are discussed within the framework of a concept of permanent alternative site oscillation of the transketolase molecule in the presence and in the absence of substrate as a manifestation of a 'flip-flop' mechanism. PMID- 9862431 TI - A putative kinetic model for substrate metabolisation by Drosophila acetylcholinesterase. AB - Insect acetylcholinesterase, an enzyme whose catalytic site is located at the bottom of a gorge, can metabolise its substrate in a wide range of concentrations (from 1 microM to 200 mM) since it is activated at low substrate concentrations. It also presents inhibition at high substrate concentrations. Among the various rival kinetic models tested to analyse the kinetic behaviour of the enzyme, the simplest able to explain all the experimental data suggests that there are two sites for substrate molecules on the protein. Binding on the catalytic site located at the bottom of the gorge seems to be irreversible, suggesting that each molecule of substrate which enters the active site gorge is metabolised. Reversible binding at the peripheral site of the free enzyme has high affinity (2 microM), suggesting that this binding increases the probability of the substrate entering the active site gorge. Peripheral site occupation decreases the entrance rate constant of the second substrate molecule to the catalytic site and strongly affects the catalytic activity of the enzyme. On the other hand, catalytic site occupation lowers the affinity of the peripheral site for the substrate (34 mM). These effects between the two sites result both in apparent activation at low substrate concentration and in general inhibition at high substrate concentration. PMID- 9862432 TI - Enhanced secretion of hydrophobic peptide fused lysozyme by the introduction of N glycosylation signal and the disruption of calnexin gene in Saccharomyces cerevisiae. AB - The insertion of a hydrophobic pentapeptide (Phe-Phe-Val-Ala-Pro) into the C terminus in hen egg white lysozyme by genetic modification resulted in an unstable structure which caused little secretion in a yeast expression system, although this modification is useful to enhance bactericidal action to gram negative bacteria [Ibrahim et al. (1994) J. Biol. Chem. 269, 5059-5063]. To enhance the secretion of the unstable hydrophobic pentapeptide fused lysozymes (H5-Lz), we attempted to introduce the signal sequence (Asn-X-Ser/Thr) of N linked glycosylation into lysozyme and to suppress the quality control of the unstable mutant in the yeast expression system. The polymannosyl hydrophobic fused lysozyme (H5/G49N-Lz) having the N-glycosylation signal sequence was expressed in the medium at 3.4 times that of unglycosylated lysozyme. Further, the secretion of the unstable mutant lysozyme was done in the Saccharomyces cerevisiae disrupted calnexin gene to avoid the degradation of the unstable mutant by the quality control. Although disruption of the calnexin gene did not lead to gross effects on the levels of growth of S. cerevisiae (W303-1b), the secretion amount of H5/G49N-Lz in calnexin disrupted S. cerevisiae was 2.5 times larger than that in wild type S. cerevisiae. These results suggest that the secretion of unstable glycosylated lysozyme (H5/G49N) was suppressed by the quality control function of calnexin and that the disruption of calnexin is effective to increase the secretion of unstable glycosylated protein. PMID- 9862433 TI - Metal coordination of azurin in the unfolded state. AB - 1H NMR data applied to the paramagnetic cobalt(II) derivative of azurin from Pseudomonas aeruginosa have made it possible to show that the metal ion is bound to the protein in the unfolded state. The relaxation data as well as the low magnetic anisotropy of the metal ion indicate that the cobalt ion is tetrahedral in the unfolded form. The cobalt ligands have been identified as the residues Gly45, His46, Cys112 and His117. Met121 is not coordinated in the unfolded state. In this state, the metal ion is not constrained to adopt a bipyramidal geometry, as imposed by the protein when it is folded. This is clear confirmation of the rack-induced bonding mechanism previously proposed for the metal ion in azurin. PMID- 9862434 TI - Constitutive transport between the trans-Golgi network and the plasma membrane according to the maturation model. A hypothesis. AB - Here we examine the application of the cisternal/carrier maturation model to describe transport of cargo proteins from the Golgi apparatus to the plasma membrane. Interpretation of the available evidence in the light of carrier maturation suggests that the transport intermediates between these stations are large pleiomorphic carriers formed by maturation of the trans-Golgi compartment, rather than vesicles, as would be postulated by the vesicular shuttle model. Mature carriers move along microtubules towards the plasma membrane via a microtubule/(kinesin)-based motor system. The maturation and vesicular transport models are compared in terms of consistency with the available literature. PMID- 9862435 TI - Involvement of the Fanconi anemia protein FA-C in repair processes of oxidative DNA damages. AB - Fanconi anemia (FA) is an autosomal recessive disorder characterized by skeletal abnormalities, pancytopenia and a marked predisposition to cancer. FA cells exhibit chromosomal instability and hypersensitivity towards oxygen and cross linking agents such as diepoxybutane and mitomycin C. An increased level of reactive oxygen intermediates and an elevation of 8-oxoguanine in FA cells point to a defective oxygen metabolism in FA cells. We investigated the repair activity of oxidatively damaged DNA in lymphoblastoid cells from FA patients of complementation groups A-E. The repair activity for oxidatively damaged DNA was significantly reduced in lymphoblastoid cell lines of complementation groups B-E. Complementation of the FA-C cell line with the wild type FA-C gene restored the repair activity to normal. This indicates that the FA-C protein participates in the repair of oxidatively damaged DNA. PMID- 9862436 TI - Identification of a functional 3',5'-cyclic adenosine monophosphate response element within the second promoter of the mouse somatostatin receptor type 2 gene. AB - Important physiological actions of somatostatin are mediated by the somatostatin receptor type 2. Its transcription is regulated by three tissue specific, alternative promoters. It is known that the mRNA of the somatostatin receptor type 2 gene is induced by cAMP, but little is known about the mechanisms underlying this regulation. We have identified and characterized a cAMP responsive element located at nucleotide -162 on the second promoter of the gene consisting of the classical palindromic octameric sequence 5'-TGACGTCA-3'. Using transient expression of reporter gene deletion constructs in NGC108-15 cells the necessity of the intact element for forskolin-induced reporter gene activity was demonstrated. The first and the third promoter are not responsive to forskolin, nor did any promoter respond to the phorbol ester PMA. Electrophoretic mobility shift assays in combination with competition experiments suggest the interaction of the promoter element with the cAMP responsive element binding protein. PMID- 9862437 TI - Probing cellular protein targets of H2O2 with fluorescein-conjugated iodoacetamide and antibodies to fluorescein. AB - Recent studies suggest that H2O2, at subtoxic concentrations generated in response to the activation of a variety of cell surface receptors, functions as an intracellular messenger. However, the intracellular targets of H2O2 action have not been identified. A procedure to detect proteins with reactive cysteine residues susceptible to oxidation by intracellularly generated H2O2 is now described. This approach is based on the labeling of proteinaceous cysteine with 5-iodoacetamidofluorescein at pH 5.5 and immunoblot analysis of the labeled proteins with antibodies specific to fluorescein. With this procedure, many proteins in human A431 cells were shown to contain reactive cysteines and to be readily oxidized by H2O2 generated in response to cellular stimulation with epidermal growth factor. One of these H2O2-sensitive proteins was identified as protein tyrosine phosphatase 1B. PMID- 9862438 TI - Obtaining and characterization of EF-hand mutants of recoverin. AB - Several EF-hand recoverin mutants were obtained and their abilities to bind to photoreceptor membranes and to inhibit rhodopsin kinase were determined. The mutants with the 'spoiled' 2nd, 3rd or (2nd+3rd) EF-hand structures did not act upon the kinase activity in the microM range of Ca2+ concentrations. Mutations of the 4th EF hand, which 'repaired' its Ca2+-binding activity, resulted in recoverin with three 'working' Ca2+-binding sites. The latter mutant inhibited rhodopsin kinase even more effectively than the wild-type recoverin, containing two working Ca2+-binding structures. PMID- 9862439 TI - An upstream negative regulatory element in human granulocyte-macrophage colony stimulating factor promoter is recognised by AP1 family members. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine involved in haematopoiesis and host defence. Production of GM-CSF has been detected in tumour cells including the U87MG astrocytoma cell line. Previous studies have been focused on the regulatory role of the proximal region of the GM-CSF promoter. Our studies on the distal region of the promoter in U87MG cells identify a negative cis element (-1377/-1298) which contains a AP1-like site able to bind c-jun and c-fos transcription factors, according to the results of DNA/protein binding assays. Mutagenesis of the AP1-like site eliminates AP1 binding and the negative effect on promoter activity. PMID- 9862441 TI - Translocation of acylated pardaxin into cells. AB - Acylated pardaxin is translocated through the cytoplasmic membrane and is accumulated in the nucleoli of NG108-15 and chromaffin cells. The uptake is time- and dose-dependent and temperature-sensitive. However, the binding of acylated 125I-pardaxin cannot be reduced by competition with pardaxin acylated with Rudinger's reagent. In this respect, acylated pardaxin resembles the Tat protein 37-71 fragment. Metabolic inhibitors do not significantly reduce the uptake of acylated 125I-pardaxin. Acylated pardaxin might be useful as a vector to translocate other molecules. PMID- 9862440 TI - Sulfonylureas blockade of neural and cardiac HERG channels. AB - The human ether-a-go-go-related gene (herg) encodes a K+ current (I(HERG)) which plays a fundamental role in heart excitability and in neurons by contributing to action potential repolarization and to spike-frequency adaptation, respectively. In this paper we show that I(HERG), recorded in neuroblastoma cells and guinea pig ventricular myocytes, was reversibly inhibited by the K(ATP) channel blocker glibenclamide (IC50 = 74 microM). The voltage and use dependence of glibenclamide blockade were also evaluated. Another sulfonylurea, glimepiride, had less effective results in blocking I(HERG). The findings of this study are relevant to the interpretation of glibenclamide effects on cellular electrophysiology and suggest that oral antidiabetic therapy with sulfonylureas may contribute to iatrogenic QT prolongation and related arrhythmias. PMID- 9862443 TI - Identification of the copper chaperone, CUC-1, in Caenorhabditis elegans: tissue specific co-expression with the copper transporting ATPase, CUA-1. AB - A cDNA encoding a putative copper chaperone protein, CUC-1, was cloned from Caenorhabditis elegans. CUC-1 had the characteristic motifs of MTCXXC and KKTGK, and showed 49.3 and 39.1% sequence identity with yeast Atx1p and human HAH1, respectively. Expression of CUC-1 cDNA complemented a null atx1 mutant, the yeast copper chaperone gene, thus demonstrating that CUC-1 is a functional copper chaperone. Studies with transgenic worms indicated that cuc-1 and cua-1, which encodes the copper transporting ATPase, are expressed together in intestinal cells of adult and hypodermal cells in the larvae. cua-1 was also expressed in pharyngeal muscle but cuc-1 was not. These results suggest that CUC-1 and CUA-1 constitute a copper trafficking pathway similar to the yeast counterparts in intestinal and hypodermal cells, and CUA-1 may have a different function in pharyngeal muscle. PMID- 9862442 TI - A phage-based system to select multiple protein-protein interactions simultaneously from combinatorial libraries. AB - Selectively infective phage (SIP) can be used to identify protein-protein interactions. SIP was modified to facilitate the simultaneous selection of interacting protein pairs from large combinatorial libraries. An interference resistant phage was constructed which non-covalently, but stably links the genetic information of an interacting pair, encoded separately on phage and phagemid vectors, by co-packaging into heteropolyphages. In a model system, the interaction between a SIP-selected peptide and the intracellular domain of the p75 neurotrophin receptor was detected in the presence of a 10(4)-fold excess of a non-interacting control pair (jun leucine zipper and p75 intracellular domain) via SIP hetero-polyphage transductants. To minimize the redundancy of transductants and to minimize possible ligand exchange generated in a solution based SIP screening, a filter-based in situ infectivity screening was developed. The combination of the above techniques may provide a powerful system for rapid screening of very large sequence spaces. PMID- 9862445 TI - A new quantitative criterion to distinguish between alpha/beta and alpha+beta proteins (domains). AB - According to the statistical analysis, it is shown that the differences of the content of alpha-helix and beta-strand between alpha/beta and alpha+beta proteins are of statistical significance. Based on the secondary structure content and the percentage of parallel or anti-parallel strands, any mixed alphabeta protein can be represented by a point in a three-dimensional prism. The distribution of the mapping points for 79 mixed alphabeta proteins (domains), of which 26 are class alpha/beta and 53 are class alpha+beta, shows that the two kinds of points are situated at distinct regions roughly. A new quantitative criterion based on the Fisher discriminant algorithm is proposed to distinguish between the alpha/beta and alpha+beta proteins (domains). Of the 79 proteins 77 are correctly classified (97.5%). As a stringent cross-validation test, the jackknife test shows that of the 79 proteins 77 are correctly classified. The jackknife test accuracy is still 97.5%. These figures indicate the self-consistence and the extrapolating effectiveness of the new quantitative criterion. Applying the new criterion to reclassify the alpha/beta and alpha+beta proteins (domains) in SCOP is also discussed. It is hoped that the new quantitative criterion will be useful for the development of protein classification databases. PMID- 9862444 TI - Expression and characterization of PP7, a novel plant protein Ser/Thr phosphatase distantly related to RdgC/PPEF and PP5. AB - We have recently identified an Arabidopsis thaliana cDNA encoding a putative protein Ser/Thr phosphatase PP7, not closely related to any protein phosphatases in animals or fungi. Here, we describe the characterization of PP7 expressed in a bacterial system. The recombinant protein was inactive unless the longest insert in its catalytic domain was cleaved, suggesting that this insert is an autoinhibitory region. PP7 was resistant to okadaic acid, calyculin and fumonisin B1, and was stimulated by Mn2+ or Fe2+, while Ni2+ and Zn2+ were inhibitory. Polylysine stimulated PP7 activity towards p-nitrophenylphosphate but inhibited activity towards the most efficient protein substrate, myelin basic protein. A tentative model of the control of PP7 activity is proposed. PMID- 9862446 TI - Identification of a retinoic acid responsive aldoketoreductase expressed in HL60 leukaemic cells. AB - Neutrophil and monocyte differentiation can be induced in HL60 leukaemia cells by all-trans-retinoic acid (ATRA) and 1alpha,25-dihydroxyvitamin D3 (D3), respectively, whose differentiating effects can be enhanced by exposure to 'anti inflammatory agents' and steroids. We have provided evidence that this potentiation is via inhibition of the activity of an enzyme of the aldoketoreductase (AKR) family, but had failed to identify expression of known AKRs in HL60 cells. In this study, we have identified a previously unclassified aldoketoreductase family member (termed HAKR e) that is expressed in HL60 cells. HAKR e is dramatically and transiently up-regulated in HL60 cells within 24 h of exposure to ATRA, further supporting the proposition that a member(s) of this family of enzymes play(s) a role in controlling cell growth and/or differentiation. PMID- 9862447 TI - Negative regulation of interleukin-1beta-activated neutral sphingomyelinase by protein kinase C in rat mesangial cells. AB - Endogenous ceramide is produced by the action of acidic or neutral sphingomyelinases (SMase) in response to stimuli such as proinflammatory cytokines or other inducers of stress. Interleukin-1beta (IL-1beta) is known to stimulate ceramide formation in rat renal mesangial cells; however, the respective subtype of SMase and its regulation have not been investigated. We found that IL-1beta induced an increase in endogenous ceramide levels via the action of a neutral SMase but not an acidic SMase in rat mesangial cells. Cytokine-induced activation of neutral SMase was inhibited by stimulation of protein kinase C (PKC) by the phorbol ester TPA which caused a reduction of ceramide back to control levels. This inhibitory effect of TPA was reversed by the specific PKC-inhibitor Ro-318220. Long-term incubation (24 h) of mesangial cells with TPA, which downregulates PKC-alpha, -delta, and -epsilon isoenzymes, resulted in a recovery of IL-1beta-stimulated neutral SMase activity as well as ceramide formation. These data implicate an important modulatory function of PKC in ceramide production in IL-1beta-activated mesangial cells. PMID- 9862448 TI - Mg2+ modulates membrane sphingolipid and lipid second messenger levels in vascular smooth muscle cells. AB - In vitro studies with smooth muscle cells from rat aorta and dog cerebral blood vessels indicate that variation in free Mg2+, within the pathophysiological range of Mg2+ concentrations, found in human serum, causes sustained changes in membrane phospholipids and lipid second messengers. Incorporation of [3H]palmitic acid into phosphatidylcholine (PC) and sphingomyelin (SM) was altered within 15 30 min after modifying the extracellular Mg2+ ion level ([Mg2+]o). Decreased Mg2+ produced a fall in both [3H]SM and [3H]PC over the first 2 h. After an 18-h incubation, the [3H]PC/[3H]SM ratio changed from about 20:1 to about 50:1. Increased [Mg2+]o resulted in a 2- to 3-fold increase in [3H]SM compared to only a small increase in [3H]PC over the same period. There was a reciprocal relationship between [3H]ceramide and [3H]1,2-DAG levels with highest [3H]ceramide and lowest [3H]-1,2-DAG levels seen at lowest [Mg2+]o. The results indicate that a fall in extracellular ionized Mg2+ concentration produces a rapid and sustained decrease in membrane sphingomyelin and a moderate rise in intracellular ceramide. A major effect of lowering [Mg2+]o appears to be a down regulation of SM synthase. The increased membrane SM content and a concomitant decrease in cell ceramide, in the presence of elevated [Mg2+]o, may be relevant to the apparent protective role of adequate Mg intake on vascular function in humans. PMID- 9862449 TI - Differential inhibition of transcription from sigma70- and sigma32-dependent promoters by rifampicin. AB - Rifampicin is an antibiotic which binds to the beta subunit of prokaryotic RNA polymerases and prevents initiation of transcription. It was found previously that production of heat shock proteins in Escherichia coli cells after a shift from 30 degrees C to 43 degrees C is not completely inhibited by this antibiotic. Here we demonstrate that while activity of a pL-lacZ fusion (pL is a sigma70 dependent promoter) in E. coli cells is strongly inhibited by rifampicin, a p(groE)-lacZ fusion, whose activity is dependent on the sigam32 factor, retains significant residual activity even at relatively high rifampicin concentrations. Differential sensitivity to this antibiotic of RNA polymerase holoenzymes containing either the sigma70 or the sigma32 subunit was confirmed in vitro. Since the effects of an antibiotic that binds to the beta subunit can be modulated by the presence of either the sigma70 or the sigma32 subunit in the holoenzyme, it is tempting to speculate that binding of various sigma factors to the core of RNA polymerase results in different conformations of particular holoenzymes, including changes in the core enzyme. PMID- 9862450 TI - Characterization of the wound-induced metallocarboxypeptidase inhibitor from potato. cDNA sequence, induction of gene expression, subcellular immunolocalization and potential roles of the C-terminal propeptide. AB - A partial cDNA clone for the potato wound-inducible metallocarboxypeptidase inhibitor (PCI) was isolated from a cDNA library constructed from mRNA of abscisic acid (ABA)-treated potato leaves. The full 5' region of the cDNA was obtained through a RACE-PCR protocol. PCI mRNA encodes a precursor polypeptide which comprises a 29 residue N-terminal signal peptide, a 27 residue N-terminal pro-region, the 39 residue mature PCI protein, and a 7 residue C-terminal extension. Northern blot analysis demonstrates that the PCI gene is transcriptionally activated by wounding, and wound signaling can be induced by ABA and jasmonic acid. Subcellular localization of the protein was investigated by immunocytochemistry and electron microscopy, showing that PCI accumulates within the vacuole. A partial PCI precursor form, comprising the mature protein and the C-terminal extension, has been expressed in Escherichia coli and characterized. Its inability to inhibit carboxypeptidases, and stability to carboxypeptidase digestion, suggest that the C-terminal pro-domain may have, besides a probable vacuolar sorting function, a role in modulation of the inhibitory activity of PCI. PMID- 9862451 TI - The effects of the Rho-kinase inhibitor Y-27632 on arachidonic acid-, GTPgammaS-, and phorbol ester-induced Ca2+-sensitization of smooth muscle. AB - The effects of the Rho-kinase inhibitor, Y-27632 [1] on Ca2+-sensitization of force induced by arachidonic acid (AA), phorbol 12,13-dibutyrate (PDBu), GTPgammaS, and by the stable thromboxane analog, 9,11-dideoxy-9alpha,11alpha methanoepoxy-PGF2alpha (U-46619), were determined in alpha-toxin-permeabilized smooth muscles. Y-27632 relaxed (up to 99%) Ca2+-sensitization by GTPgammaS (10 microM) and U46619 (1 microM), but not by PDBu (20 microM), and reduced GTPgammaS induced myosin light chain (MLC20) phosphorylation from 28% to 17% (P=0.002). GTPgammaS-induced force sensitization was inhibited by Y-27632 more potently when the inhibitor was added during the plateau of force than prior to stimulation. In alpha-toxin-permeabilized smooth muscle, Y-27632 inhibited AA (50 microM)-induced Ca2+-sensitization of force (by 66 +/- 1.3%) and reduced MLC20 phosphorylation. In contrast, Y-27632 did not relax force Ca2+-sensitized by AA in smooth muscle permeabilized with Triton X-100. We conclude that (i) AA induces Ca2+ sensitization through dual mechanisms, one mediated by Rho-kinase (or a related kinase), and (ii) Rho-kinase is not required for phorbol ester-induced Ca2+ sensitization. PMID- 9862452 TI - Molecular cloning, functional characterization and mRNA expression analysis of the murine chemokine receptor CCR6 and its specific ligand MIP-3alpha. AB - We have cloned the murine CCR6 receptor and its ligand, the beta-chemokine mMIP 3alpha. Calcium mobilization assays performed with mCCR6 transfectants showed significant responses upon addition of mMIP-3alpha. Murine MIP-3alpha RNA is expressed in thymus, small intestine and colon, whereas mCCR6 RNA is expressed in spleen and lymph nodes. RT-PCR analysis of FACS-sorted lymphoid and antigen presenting cell subsets showed mCCR6 expression mainly in B cells, CD8- splenic dendritic cells and CD4+ T cells. The cloning and functional characterization of the mCCR6 and mMIP-3alpha will allow the study of the role of these proteins in mouse models of inflammation and immunity. PMID- 9862453 TI - Lack of effect of RPE65 removal on the enzymatic processing of all-trans-retinol into 11-cis-retinol in vitro. AB - RPE65 is a major membrane associated protein found in the vertebrate retinal pigment epithelium (RPE). Various studies have shown this protein to be essential for visual function, possibly at the level of the processing of retinoids. The pigment epithelium is the anatomical site in which the visual chromophore 11-cis retinal is generated. The two critical RPE enzymes in the isomerization pathway are lecithin retinol acyl transferase (LRAT) and isomerohydrolase, which processes all-trans-retinyl esters into 11-cis-retinol. Both enzymes are membrane bound. It is shown here that RPE65 can be largely extracted (90-95%) from RPE membranes by 1 M KCl by itself, or with added detergent CHAPS. The almost quantitative extraction of RPE65 from RPE membranes has little or no effect on in vitro LRAT and isomerohydrolase activities in quantitative enzymatic assays using RPE membranes, suggesting that RPE65 may not have an important role to play in the enzymatic processing of all-trans-retinol into 11-cis-retinol in vitro. PMID- 9862454 TI - Extracellular proton alters the divalent cation binding affinity in a cyclic nucleotide-gated channel pore. AB - Extracellular protons in the range of 10(-9) to 10(-5) M effectively suppressed Na+ current (K(1/2) = 10(-6.1)) through the bovine retinal guanosine 3',5'-cyclic mononucleotide-gated ion channel expressed in Xenopus oocytes. The reduction of channel current was mediated by a single glutamate residue (Glu363) within the pore-forming region of the channel, also involved in extracellular divalent cation binding. Increasing the concentration of extracellular proton decreased the binding affinity of the extracellular divalent cation (e.g. Sr2+) and the large difference of binding affinity previously observed between the wild-type and E363D mutant channel disappeared. These results indicate that the permeation characteristics of cyclic nucleotide-gated ion channel can be altered by extracellular pH through a single acidic residue in the channel conduction pathway. PMID- 9862455 TI - Putative helices 3 and 5 of the human vitamin D3 receptor are important for the binding of calcitriol. AB - We have introduced eleven point mutations into the human vitamin D receptor by site-directed mutagenesis in order to identify some of the amino acid residues that are important for ligand binding. The amino acid residues Ser225, His229, Asp232, Val234, Ser235, Tyr236, Ser237, Lys240, Ile242, Lys246 (helix 3), and Ser275 (helix 5) of the human vitamin D receptor were substituted by alanine. We report here that His229, Asp232, and Ser237 have an important role in the binding of calcitriol. In addition, the amino acid residues Tyr236 and Ser275 also seem to participate in the ligand binding process. PMID- 9862456 TI - From beta-glucanase to beta-glucansynthase: glycosyl transfer to alpha-glycosyl fluorides catalyzed by a mutant endoglucanase lacking its catalytic nucleophile. AB - Removal of the catalytic nucleophile Glu134 of the retaining 1,3-1,4-beta glucanase from Bacillus licheniformis by mutation to alanine yields an enzyme with no glycosidase activity. The mutant is able to catalyze the regio- and stereospecific glycosylation of alpha-laminaribiosyl fluoride with different glucoside acceptors through a single-step inverting mechanism. The main advantage of the mutant as glycosylation catalyst with respect to the kinetically controlled transglycosylation using the wild-type enzyme is that the reaction products cannot be hydrolyzed by the mutant enzyme, and glycosylation yields rise to 90%. PMID- 9862457 TI - Bacteriophage lambda surface display of a bacterial biotin acceptor domain reveals the minimal peptide size required for biotinylation. AB - Phage display is a powerful technique for identifying specific ligands to a given target. In this work random peptides derived from the biotin accepting domain of the Klebsiella pneumoniae oxaloacetate decarboxylase were displayed on bacteriophage lambda heads to determine the minimal sequence length that is necessary to effect biotinylation in vivo. Phages with a functional biotinylation domain were identified after affinity purification with immobilised avidin. All biotinylated phages isolated this way were found to have a sequence of 66 amino acids from the parental protein in common. This minimal biotinylation domain is fully functional as a biotin acceptor and more resistant to proteolytic attack compared to domains of larger size derived from the same protein. The data present the first example of a posttranslational protein modification analysed in a phage display system. Moreover, a biotin domain of reduced size and improved stability was identified, that should be superior to the larger parental protein as a tag to generate biotinylated fusion proteins. PMID- 9862458 TI - Inhibition of skeletal muscle sarcoplasmic reticulum Ca2+-ATPase by nitric oxide. AB - The effects of nitric oxide on the activities of thapsigargin-sensitive sarcoplasmic reticulum Ca2+-ATPase (SERCA) and Ca2+ uptake by sarcoplasmic reticulum (SR) membranes prepared from white skeletal muscle of rabbit femoral muscle were studied. Pretreatment of the SR preparations with nitric oxide at concentrations of up to 250 microM for 1 min decreased the SERCA activity concentration dependently, and also decreased their Ca2+ uptake. Both these effects of nitric oxide were reversible. Inhibitors of guanylyl cyclase and protein kinase G (PKG) had no significant effect on the nitric oxide-induced inhibitions of SERCA and Ca2+ uptake. Moreover, dithiothreitol did not reverse the inhibitory effects of nitric oxide on SERCA and Ca2+ uptake. These findings suggest that nitric oxide inhibits SERCA, mainly SERCA 1, of rabbit femoral skeletal muscle by an action independent of the cyclic GMP-PKG system or oxidation of thiols, and probably by a direct action on SERCA protein. PMID- 9862459 TI - Detection of the local H+ gradients on the internal mitochondrial membrane. AB - Respiration-dependent responses of a pH probe (fluorescein isothiocyanate, FITC), covalently bound to the membrane proteins of mitochondria and submitochondrial particles (SMP) have been studied. A spectral shift indicating FITC deprotonation was observed when respiration was activated in coupled mitochondria. Such a response was increased by valinomycin and reduced by uncoupler. Some FITC deprotonation was detected in the presence of excess of an uncoupler, but the response was smaller and insensitive to valinomycin. FITC deprotonation was also observed in submitochondrial particles after succinate addition. In this case it was not affected by uncoupler. Increase in the buffer concentration was found to (i) decrease the FITC response and (ii) increase the rate of uncoupled respiration in both mitochondria and submitochondrial particles. The results are consistent with the assumption that respiration initiates appearance of local H+ activity gradients on the inner side of the internal mitochondrial membrane during the steady-state H+ pumping. We suggest that the formation of this gradient is due to kinetic barrier to proton transfer from the bulk phase to the respiratory proton pump vicinity. PMID- 9862460 TI - Biotin synthase mechanism: on the origin of sulphur. AB - Biotin synthase catalyses the last step of the biosynthesis of biotin in microorganisms and plants. The active protein isolated from Bacillus sphaericus and Escherichia coli contains an iron-sulphur (FeS) cluster. The native enzymes were depleted of their iron and inorganic sulphide and the resulting apoenzymes were chemically reconstituted with FeCl3 and Na2[34S] to give labelled (Fe34S) enzymes. These enzymes were functional and when assayed in vitro produced labelled biotin containing about 65% of 34S. These data strongly support the hypothesis that the sulphur of biotin is derived from the (FeS) centre of the enzyme. PMID- 9862461 TI - Expression of an oncogenic mutant G alpha i2 activates Ras in Rat-1 fibroblast cells. AB - It has been reported that expression of the active mutant of heterotrimeric GTP binding protein alpha subunit G alpha i2 transforms Rat-1 cells. However, the G alpha i2-mediated mitogenic signaling pathways remain to be elucidated. Here, we demonstrate that inducible expression of the active mutant of G alpha i2 (G alpha i2(Q205L)) activates Ras and c-Jun N-terminal kinase (JNK) in addition to extracellular signal-regulated kinase (ERK) in Rat-1 cells. Our findings suggest that Ras may play a critical role in the G alpha i2-induced transformation and G alpha i2 can transduce signals from the Gi-coupled receptor to JNK and ERK in certain types of mammalian cells. PMID- 9862462 TI - Polylysine decelerates kinetics of negatively charged gramicidin channels as shown by sensitized photoinactivation. AB - Effect of a cationic polymer, poly(L-lysine), on the kinetic properties of ionic channels formed by neutral gramicidin A (gA) and its negatively charged analogue O-pyromellitylgramicidin (OPg) in a bilayer lipid membrane is studied using a method of sensitized photoinactivation. This newly developed method is based on the analysis of transmembrane current transients induced by a flash in the presence of a photosensitizer. It has been shown previously that the time course of the flash-induced current decrease in most cases follows a single exponential decay with an exponential factor (tau, the characteristic time of photoinactivation) that correlates well with the single-channel lifetime. Addition of polylysine does not affect tau for gA channels, but causes a substantial increase in tau for OPg channels. This effect is reversed by addition of polyacrylic acid. The deceleration of the photoinactivation kinetics is ascribed to electrostatic interaction of polylysine with OPg probably resulting in OPg clustering. The latter can stabilize the channel state by reducing the rotational and lateral mobility of OPg monomers and dimers, and thus increase the single channel lifetime. PMID- 9862463 TI - Identification of the three non-identical subunits constituting human deoxyribonuclease II. AB - We purified DNase II from human liver to apparent homogeneity. The N-terminal amino acid sequences of each of three components constituting the purified mature enzyme were then separately determined by automatic Edman degradation. A combination of this chemical information and the previously reported nucleotide sequence of the cDNA encoding human DNase II [Yasuda et al. (1998) J. Biol. Chem. 273, 2610-2626] allowed detailed elucidation of the enzyme's subunit structure: human DNase II was composed of three non-identical subunits, a propeptide, proprotein and mature protein, following a signal peptide. Expression analysis of a series of deletion mutants derived from the cDNA of DNase II in COS-7 cells suggested that although a single large precursor protein may not be necessary for proteolytic maturation, the propeptide region L17-Q46 may play an essential role in generating the active form of the enzyme. PMID- 9862464 TI - The Drosophila inhibitor of apoptosis D-IAP1 suppresses cell death induced by the caspase drICE. AB - Many members of the Inhibitor of Apoptosis (IAP) family inhibit cell death and existing data suggest at least two mechanisms of action. Drosophila IAPs (D-IAP1 and D-IAP2) and a baculovirus-derived IAP, Op-IAP, physically interact with and inhibit the anti-apoptotic activity of Reaper, HID, and Grim, three genetically defined inducers of apoptosis in Drosophila, while human IAPs, c-IAP1, c-IAP2, and X-IAP interact with a number of different proteins including specific members of the caspase family of cysteine proteases which are crucial in the execution of cell death. We have examined whether insect-active IAPs can inhibit apoptosis induced by selected caspases, Drosophila drICE, Sf-caspase-1, and mammalian caspase-3, in insect SF-21 cells. D-IAP1 inhibited apoptosis induced by the active forms of all three caspases tested and physically interacted with the active, but not the proform of drICE. MIHA, the mouse homolog of X-IAP and an effective inhibitor of caspase-3, also interacted with and blocked apoptosis induced by active drICE but was relatively ineffective in blocking Sf-caspase-1. Op-IAP and D-IAP2 were unable to inhibit effectively any of the active caspases tested and failed to interact with drICE. The Drosophila IAPs and Op-IAP, but not MIHA, blocked HID-initiated activation of pro-drICE. We conclude that D-IAP1 is capable of inhibiting the activation of drICE as well as inhibiting apoptosis induced by the active form of drICE. In contrast, D-IAP2 and Op-IAP are more limited in their inhibitory targets and may be limited to inhibiting the activation of caspases. PMID- 9862465 TI - The sugarless mutation affects the expression of the white eye color gene in Drosophila melanogaster. AB - Investigation of a modifier locus displaying a darker eye phenotype in white apricot flies led to the isolation of the gene encoding UDP-glucose dehydrogenase (UDPGDH). The P-element insertion l(3)05007 occurs upstream of the transcription start site of the sugarless (sgl) gene and greatly reduces its transcription at various developmental stages. A single abundant sgl transcript shows a ubiquitous distribution and encodes a 53-kDa protein which is 64% identical in sequence to bovine UDP-glucose dehydrogenase. Overexpression of sgl in E. coli resulted in synthesis of a protein with high levels of UDPGDH activity. Expression of three genes that participate in pigment deposition, white, scarlet and brown, was significantly affected in populations segregating for sgl, suggesting that it is the decrease in UDPGDH level that produces the modifying effect observed. In addition, genetic effects on white-apricot were observed in sgl-wingless and sgl hedgehog double mutants. Recent data have indicated an effect of UDPGDH on cell surface glycosaminoglycans (GAGs) which modulate the activity of growth factors, and in particular wingless signaling. Our results suggest that the levels of GAGs are rate limiting for cell-cell signaling pathways which mediate changes in gene expression. PMID- 9862467 TI - In vivo linearization and autonomous replication of plasmids containing human telomeric DNA in Aspergillus nidulans. AB - Plasmids containing two inverted 0.6-kb stretches of human telomeric repeats transform Aspergillus nidulans at frequencies characteristic of autonomously replicating vectors. Transformation frequency is not affected when the plasmids are linearized in vitro prior to transformation by cutting between the inverted repeats. Southern analysis reveals the presence of a homogeneous pool of linear plasmid molecules in mycelium of transformants. Addition of the AMA1 plasmid replicator to the telomere-containing plasmids has only a minor effect on transformation. The phenotypic stability of the transformants is low. However, unlike conventional replicative transformants containing AMA1-bearing plasmids, these transformants are prone to spontaneous stabilization which occurs predominantly by conversion of the mutant chromosomal allele of the marker gene to the plasmid-borne allele. The data strongly suggest that telomeric DNA can act as a plasmid replicator. An alternative interpretation is that autonomous replication of linear DNA fragments, in contrast to covalently closed supercoiled molecules, does not require any special replicator sequences. PMID- 9862466 TI - The gene cluster inlC2DE of Listeria monocytogenes contains additional new internalin genes and is important for virulence in mice. AB - In this work we identified and characterized a gene cluster containing three internalin genes of Listeria monocytogenes EGD. These genes, termed inlG, inlH and inlE, encode proteins of 490, 548 and 499 amino acids, respectively, which belong to the family of large, cell wall-bound internalins. The inlGHE gene cluster is flanked by two listerial house-keeping genes encoding proteins homologous to the 6-phospho-beta-glucosidase and the succinyl-diaminopimelate desuccinylase of E. coli. A similar internalin gene cluster, inlC2DE, localised to the same position on the L. monocytogenes EGD chromosome was recently described in a different isolate (Dramsi S, Dehoux P, Lebrun M, Goossens PL, Cossart P (1997) Infect Immun 65: 1615-1625). Sequence comparison of the two inl gene clusters indicates that inlG is a new internalin gene, while inlH was generated by a site-specific recombination, leading to an in-frame deletion which removed the 3'-terminal end of inlC2 and the 5'-terminal part of inlD. The third gene of the inlGHE cluster, inlE, is almost identical to the previously reported inlE gene. Our data show that the inlGHE gene cluster is probably transcribed from a major PrfA-independent promoter located upstream of inlG. PCR analysis revealed the presence of the newly identified inl genes inlG and inlH in most L. monocytogenes isolates tested. A mutant which has lost inlG, inlH and inlE by an in-frame deletion exhibited, after oral infection of mice, a significant loss in virulence and shows drastically reduced numbers of viable bacteria in both liver and spleen when compared to the wild-type strain. PMID- 9862468 TI - Duplication of the Rdl GABA receptor subunit gene in an insecticide-resistant aphid, Myzus persicae. AB - Resistance to cyclodiene insecticides is associated with replacements of a single amino acid (alanine 302) in a gamma-aminobutyric acid (GABA) receptor subunit encoded by the single-copy gene Resistance to dieldrin (Rdl). Alanine 302 is predicted to reside within the second membrane-spanning region of the Rdl receptor, a region that is thought to line the integral chloride ion channel pore. In all cyclodiene-resistant insects studied to date, this same alanine residue is replaced either by a serine, or, in some resistant strains of Drosophila simulans, a glycine residue. Therefore, individuals can carry only two different Rdl alleles. In contrast, here we report the presence of up to four different Rdl-like alleles in individual clones of the green peach aphid, Myzus persicae. In addition to the wild-type copy of Rdl gene (encoding A302 or allele A), M. persicae carries three other alleles with the following amino acid replacements: A302-->Glycine (allele G), A302-->SerineTCG (allele S) and A302- >SerineAGT (allele S'). Evidence from direct nucleotide sequencing and Single Stranded Conformational Polymorphism (SSCP) analysis shows that at least three of these different Rdl alleles (i.e. A, G and S) are commonly present in individual aphids or aphid clones. Southern analysis using allele-specific probes and analysis of sequences downstream of the exon containing the resistance-associated mutation confirm the presence of two independent Rdl-like loci in M. persicae. One locus carries the susceptible alanine (A) and/or resistant glycine (G) allele while the other carries the two serine alleles (S or S'). Whereas resistance levels are correlated with the glycine replacement, the S allele was present in all aphid clones, regardless of their resistance status. These results suggest that target site insensitivity is associated with replacements at the first (A/G) but not the second (S/S') locus. Phylogenetic analysis of nucleotide sequences indicates that both putative aphid Rdl loci are monophyletic with respect to other insect Rdl genes and may have arisen through a recent gene duplication event. The implications of this duplication with respect to insecticide resistance and insect GABA receptor subunit diversity are discussed. PMID- 9862469 TI - mRNA surveillance mitigates genetic dominance in Caenorhabditis elegans. AB - Nonsense mutant mRNAs are unstable in all eucaryotes tested, a phenomenon termed nonsense-mediated mRNA decay (NMD) or mRNA surveillance. Functions of the seven smg genes are required for mRNA surveillance in Caenorhabditis elegans. In Smg(+) genetic backgrounds, nonsense-mutant mRNAs are unstable, while in Smg(-) backgrounds such mRNAs are stable. Previous work has demonstrated that the elevated level of nonsense-mutant mRNAs in Smg(-) animals can influence the phenotypic effects of heterozygous nonsense mutations. Certain nonsense alleles of a muscle myosin heavy chain gene are recessive in Smg(+) backgrounds but strongly dominant in Smg(-) backgrounds. Such alleles probably express disruptive myosin polypeptide fragments whose abundance is elevated in smg mutants due to elevation of mRNA levels. We report here that mutations in a variety of C. elegans genes are strongly dominant in Smg(-), but recessive or only weakly dominant in Smg(+) backgrounds. We isolated 32 dominant visible mutations in a Smg(-) genetic background and tested whether their dominance requires a functional NMD system. The dominance of 21 of these mutations is influenced by NMD. We demonstrate, furthermore, that in the case of myosin, the dominant negative effects of nonsense alleles are likely to be due to expression of N terminal nonsense-fragment polypeptides, not to mistranslation of the nonsense codons. mRNA surveillance, therefore, may mitigate potentially deleterious effects of many heterozygous germline and somatic nonsense or frame-shift mutations. We also provide evidence that smg-6, a gene previously identified as being required for NMD, performs essential function(s) in addition to its role in NMD. PMID- 9862470 TI - A cluster of charged and aromatic residues in the C-terminal portion of maltoporin participates in sugar binding and uptake. AB - The maltoporin LamB of Escherichia coli K12 is a trimeric protein which facilitates the diffusion of maltose and maltodextrins through the bacterial outer membrane, and also acts as a non-specific porin for small hydrophilic molecules as well as a receptor for phages. Loop L9 (residues 375 to 405) is the most distal and largest surface-exposed loop of LamB. It comprises a central portion, which varies in size and sequence in the maltoporins of known sequence, flanked by two conserved regions containing charged and aromatic residues. In order to identify the residues within the proximal region that are specifically involved in sugar utilization, we used site-directed mutagenesis to change, individually, each of the charged (five) and aromatic (three) residues in the region 371 to 379 into alanine. None of the eight single amino acid substitutions affected the phage receptor activity of LamB. In contrast, they all affected, to variable extents, maltoporin functions. For all the mutants, very good correlations were observed between the effects on sugar binding and on in vivo uptake. In no case were maltoporin functions completely abolished. Mutants E374 A and W376 A were the most impaired (with over 60% reduction in dextrin binding and in vivo uptake of maltose and maltopentaose). These two mutations also led to an increased bacterial sensitivity to bacitracin and vancomycin. The functional and structural implications are discussed. PMID- 9862471 TI - Crosstalk between cAMP and pheromone signalling pathways in Ustilago maydis. AB - In the phytopathogenic basidiomycete Ustilago maydis mating and dikaryon formation are controlled by a pheromone/receptor system and the multiallelic b locus. Recently, a gene encoding a G protein alpha subunit, gpa3, was isolated and has subsequently been implicated in pheromone signal transduction. Mutants deleted for gpa3 are sterile and nonpathogenic, and exhibit a morphology that is similar to that of mutants with defects in the adenylate cyclase gene uac1. We have found that the sterility and mutant morphology of gpa3 deletion strains can be rescued by exogenous cAMP. In these mutants and in the corresponding wild-type strains, exogenous cAMP stimulates pheromone gene expression to a level comparable to that seen in the pheromone-stimulated state. In addition, we demonstrate that uac1 is epistatic to gpa3. We conclude that Gpa3 controls the cAMP signalling pathway in U. maydis and discuss how this pathway feeds into the pheromone response. PMID- 9862472 TI - DnaK-dependent ribosome biogenesis in Escherichia coli: competition for dominance between the alleles dnaK756 and dnaK+. AB - The Escherichia coli chaperone DnaK is vital for many cellular functions, including ribosome biogenesis at high temperature. Thus, the dnaK756-ts (lambdaR) mutant, at the non-permissive temperature, is inhibited at a late stage of ribosome assembly, yielding 21S, 32S and 45S precursor particles. This defect, unlike the lambda resistance and thermosensitivity phenotypes, is not complemented by lysogenisation with a transducing phage lambda dnaK+ bearing the wild-type dnaK gene. However this dominant phenotype becomes recessive when dnaK+ is expressed from a medium-copy-number plasmid. On the other hand, an excess of DnaK causes an unexpected dominant-lethal effect of the dnaK756 allele near non permissive temperatures. This interplay between the dnaK+ and dnaK756 alleles supports the idea of that DnaK oligomers form in the cell. PMID- 9862473 TI - The chloroplast division protein FtsZ is encoded by a nucleomorph gene in cryptomonads. AB - Guillardia theta is a cryptomonad alga, whose phototrophic symbiont was acquired by secondary endocytobiosis. The nucleomorph, the vestigial nucleus of the eukaryotic endosymbiont, harbors three linear chromosomes with a total coding capacity of 515 kb. Sequencing of the nucleomorph genome reveals that it encodes an ORF homologous to the bacterial cell division protein FtsZ, supporting the hypothesis that FtsZ is common in chloroplasts. We show that the nucleomorph encoded ftsZ gene is transcribed. The transcript is polyadenylated and therefore shows features typical of eukaryotic transcripts. However, 3' processing of nucleomorph mRNA is inaccurate. Transcripts of nucleomorph genes in G. theta overlap with neighboring UTRs and coding regions. We demonstrate that the reading frame encoding NmFtsZ is not interrupted by introns. Subcellular localization of the protein reveals that FtsZ is localized exclusively in the chloroplast of G. theta, demonstrating that FtsZ is imported into the organelle. PMID- 9862474 TI - Cloning and characterization of the rpoH gene of Rhodobacter capsulatus. AB - By using an oligonucleotide mixture corresponding to a region highly conserved among alternative sigma factors we identified a new sigma factor gene (rpoH) from Rhodobacter capsulatus. This gene encodes a protein of 34 kDa with strong similarity to the RpoH (sigma32) factors from other bacterial species. It was not possible to inactivate the R. capsulatus rpoH gene by introducing a resistance cassette, implying that it is essential for growth. The 5' ends of the mRNAs were mapped to two sequences with similarity to an rpoH- and an rpoD-dependent promoter, respectively. The amounts of both these mRNAs increased after heat shock, but were unaffected by a decrease in oxygen tension. Western analysis using a sigma factor-specific antibody revealed the accumulation of a protein of about 34 kDa after heat shock, and an increase in the amounts of a protein with the same size after reduction of oxygen tension in R. capsulatus cultures. PMID- 9862475 TI - The expression of genes involved in parasitism by Trichoderma harzianum is triggered by a diffusible factor. AB - The mycoparasite Trichoderma harzianum has been extensively used in the biocontrol of a wide range of phytopathogenic fungi. Hydrolytic enzymes secreted by the parasite have been directly implicated in the lysis of the host. Dual cultures of Trichoderma and a host, with and without contact, were used as means to study the mycoparasitic response in Trichoderma. Northern analysis showed high level expression of genes encoding a proteinase (prb1) and an endochitinase (ech42) in dual cultures even if contact with the host was prevented by using cellophane membranes. Neither gene was induced during the interaction of Trichoderma with lectin-coated nylon fibres, which are known to induce hyphal coiling and appressorium formation. Thus, the signal involved in triggering the production of these hydrolytic enzymes by T. harzianum during the parasitic response is independent of the recognition mediated by this lectin-carbohydrate interaction. The results showed that induction of prb1 and ech42 is contact independent, and a diffusible molecule produced by the host is the signal that triggers expression of both genes in vivo. Furthermore, a molecule that is resistant to heat and protease treatment, obtained from Rhizoctonia solani cell walls induces expression of both genes. Thus, this molecule is involved in the regulation of the expression of hydrolytic enzymes during mycoparasitism by T. harzianum. PMID- 9862476 TI - Cloning and expression of the ApaLI, NspI, NspHI, SacI, ScaI, and SapI restriction-modification systems in Escherichia coli. AB - The genes encoding the ApaLI (5'-GTGCAC-3'), NspI (5'-RCATGY-3'), NspHI (5' RCATGY-3'), SacI (5'-GAGCTC-3'), SapI (5'-GCTCTTCN1-3', 5'-N4GAAGAGC-3') and ScaI (5'-AGTACT-3') restriction-modification systems have been cloned in E. coli. Amino acid sequence comparison of M.ApaLI, M.NspI, M.NspHI, and M.SacI with known methylases indicated that they contain the ten conserved motifs characteristic of C5 cytosine methylases. NspI and NspHI restriction-modification systems are highly homologous in amino acid sequence. The C-termini of the NspI and NlaIII (5'-CATG-3') restriction endonucleases share significant similarity. 5mC modification of the internal C in a SacI site renders it resistant to SacI digestion. External 5mC modification of a SacI site has no effect on SacI digestion. N4mC modification of the second base in the sequence 5'-GCTCTTC-3' blocks SapI digestion. N4mC modification of the other cytosines in the SapI site does not affect SapI digestion. N4mC modification of ScaI site blocks ScaI digetion. A DNA invertase homolog was found adjacent to the ApaLI restriction modification system. A DNA transposase subunit homolog was found upstream of the SapI restriction endonuclease gene. PMID- 9862478 TI - The MCM16 gene of the yeast Saccharomyces cerevisiae is required for chromosome segregation. AB - We have cloned and characterized the MCM16 gene required for the maintenance of minichromosomes in the yeast Saccharomyces cerevisiae. This gene corresponds to a 181-amino acid ORF, YPR046W, on chromosome XVI. Mutant cells carrying minichromosomes accumulate them in higher copy numbers than do wild-type cells. Intact dicentric plasmid could be recovered from the mutant, in contrast to the wild-type, in which the plasmid suffered frequent deletions. A wild-type centromere, CEN6, acts as a block to the transcription of a reporter gene, such as beta-galactosidase. This block was less effective in the mutant than in the wild-type strain, suggesting alterations in kinetochore assembly in the former. The mutant also showed increased sensitivity to the antimitotic drugs benomyl and thiabendazole. The mcm16 mutation caused a high rate of loss of chromosome III, without any significant increase in the recombination frequency. A strain carrying a deletion-disruption derivative of the MCM16 gene was viable and, when compared to the wild-type, did not show any significant changes in growth rate or cell morphology at 16, 23 and 37 degrees C. These properties show that MCM16 is required for an important but nonessential role that governs the kinetochore microtubule mediated process of chromosome segregation. PMID- 9862477 TI - Efficient initiation of S-phase in yeast requires Cdc40p, a protein involved in pre-mRNA splicing. AB - The S. cerevisiae CDC40 gene was originally identified as a cell-division specific gene that is essential only at elevated temperatures. Cells carrying mutations in this gene arrest with a large bud and a single nucleus with duplicated DNA content. Cdc40p is also required for spindle establishment or maintenance. Sequence analysis reveals that CDC40 is identical to PRP17, a gene involved in pre-mRNA splicing. In this paper, we show that Cdc40p is required at all temperatures for efficient entry into S-phase and that cell cycle arrest associated with cdc40 mutations is independent of all the known checkpoint mechanisms. Using immunofluorescence, we show that Cdc40p is localized to the nuclear membrane, weakly associated with the nuclear pore. Our results point to a link between cell cycle progression, pre-mRNA splicing, and mRNA export. PMID- 9862479 TI - Interallelic complementation at the pyrF locus and the homodimeric nature of orotate phosphoribosyltransferase (OPRTase) in Mucor circinelloides. AB - Using 5-fluoroorotic acid (5-FOA) as a positive selection system we isolated mutants of Mucor circinelloides altered in the pyrimidine biosynthetic pathway. These mutants were found to be deficient either in orotidine-5'-monophosphate decarboxylase (OMPdecase), or in orotate phosphoribosyltransferase (OPRTase) activity. Complementation tests among mutants lacking OPRTase activity classified them into three groups, thus suggesting the possibility of interallelic complementation. To investigate this hypothesis a cDNA clone corresponding to the OPRTase-encoding gene of M. circinelloides was isolated by direct complementation of E. coli. The genomic copy transformed to prototrophy one member of each of the three classes of OPRTase-deficient mutants. We therefore concluded that they were all altered at the same locus, the pyrF locus. The corresponding alleles were cloned and sequenced. Comparisons of the amino acid sequence of M. circinelloides OPRTase with those of E. coli and S. typhimurium revealed a high degree of similarity in secondary and tertiary structure. As the two bacterial enzymes exist as dimers, a homodimeric quaternary structure of the M. circinelloides mature protein can be assumed. This would also explain the interallelic complementation between some pyrF mutants. The mutations found could affect either the active site or the structure of the dimer interface of the OPRTase. PMID- 9862480 TI - Cloning and characterization of cDNAs encoding S-RNases from almond (Prunus dulcis): primary structural features and sequence diversity of the S-RNases in Rosaceae. AB - cDNAs encoding three S-RNases of almond (Prunus dulcis), which belongs to the family Rosaceae, were cloned and sequenced. The comparison of amino acid sequences between the S-RNases of almond and those of other rosaceous species showed that the amino acid sequences of the rosaceous S-RNases are highly divergent, and intra-subfamilial similarities are higher than inter-subfamilial similarities. Twelve amino acid sequences of the rosaceous S-RNases were aligned to characterize their primary structural features. In spite of their high level of diversification, the rosaceous S-RNases were found to have five conserved regions, C1, C2, C3, C5, and RC4 which is Rosaceae-specific conserved region. Many variable sites fall into one region, named RHV. RHV is located at a similar position to that of the hypervariable region a (HVa) of the solanaceous S-RNases, and is assumed to be involved in recognizing S-specificity of pollen. On the other hand, the region corresponding to another solanaceous hypervariable region (HVb) was not variable in the rosaceous S-RNases. In the phylogenetic tree of the T2/S type RNase, the rosaceous S-RNase fall into two subfamily-specific groups (Amygdaloideae and Maloideae). The results of sequence comparisons and phylogenetic analysis imply that the present S-RNases of Rosaceae have diverged again relatively recently, after the divergence of subfamilies. PMID- 9862481 TI - Four hydrophobic amino acid residues in the C-terminal effector domain of the yeast Mig1p repressor are important for its in vivo activity. AB - The Mig1 repressor is a zinc finger protein that mediates glucose repression in yeast. Previous work in Saccharomyces cerevisiae has shown that two domains in Miglp are required for repression: the N-terminal zinc finger region and a C terminal effector domain. Both domains are also conserved in Miglp homologs from the distantly related yeasts Kluyveromyces lactis and K. marxianus, and these Mig1 proteins can fully replace the endogenous Mig1p in S. cerevisiae. We have now made a detailed analysis of the conserved C-terminal effector domain in Mig1p from K. marxianus, using expression in S. cerevisiae to monitor its function. First, a series of small deletions were made within the effector domain. Second, an alanine scan mutagenesis was carried out across the effector domain. Third, double, triple and quadruple mutants were made that affect certain residues within the effector domain. Our results show that four conserved residues within the effector domain, three leucines and one isoleucine, are particularly important for its function in vivo. The analysis further revealed that while the C-terminal effector domain of KmMig1p mediates a seven- to nine-fold repression of the reporter gene, a five- to sixfold residual effect also exists that is independent of the C-terminal effector domain. Similar results were obtained when the corresponding mutations were made in ScMig1p. Moreover, we found that mutations in these residues affect the interaction between Mig1p and the general corepressor subunit Cyc8p (Ssn6p). Modeling of the C-terminal effector domain using a protein of known structure suggests that it may be folded into an alpha helix. PMID- 9862483 TI - The Caenorhabditis elegans RAD51 homolog is transcribed into two alternative mRNAs potentially encoding proteins of different sizes. AB - In prokaryotes, the RecA protein plays a pivotal role in homologous recombination, catalyzing the transfer of a single DNA strand into an homologous molecule. Structural homologs of the bacterial RecA protein, called Rad51, have been found in different eukaryotes (from yeast to man), suggesting a certain level of conservation in recombination pathways among living organisms. We have cloned the homolog of RAD51 in Caenorhabditis elegans. The CeRAD51 gene is transcribed into two alternative mRNAs and potentially codes for two proteins of 395 and 357 amino acids in length, respectively. We discuss the evolutionary implications of these findings. PMID- 9862482 TI - Lethal mutations defining 112 complementation groups in a 4.5 Mb sequenced region of Caenorhabditis elegans chromosome III. AB - The central gene cluster of chromosome III was one of the first regions to be sequenced by the Caenorhabditis elegans genome project. We have performed an essential gene analysis on the left part of this cluster, in the region around dpy-17III balanced by the duplication sDp3. We isolated 151 essential gene mutations and characterized them with regard to their arrest stages. To facilitate positioning of these mutations, we generated six new deficiencies that, together with preexisting chromosomal rearrangements, subdivide the region into 14 zones. The 151 mutations were mapped into these zones. They define 112 genes, of which 110 were previously unidentified. Thirteen of the zones have been anchored to the physical sequence by polymerase chain reaction deficiency mapping. Of the 112 essential genes mapped, 105 are within these 13 zones. They span 4.2 Mb of nucleotide sequence. From the nucleotide sequence data, 920 genes are predicted. From a Poisson distribution of our mutations, we predict that 234 of the genes will be essential genes. Thus, the 105 genes constitute 45% of the estimated number of essential genes in the physically defined zones and between 2 and 5% of all essential genes in C. elegans. PMID- 9862484 TI - Mouse homolog of the yeast origin recognition complex subunit ORC1 and chromosomal localization of the cognate mouse gene Orc1. AB - ORC1 encodes a subunit of the eukaryotic origin recognition complex in yeast, which has been shown to play a crucial role in chromosomal DNA replication and transcriptional silencing. We have identified a mouse protein closely related to Saccharomyces cerevisiae ORC1 as well as to the human, Xenopus and Drosophila homologs. MmORC1 has putative cyclin-dependent phosphorylation sites and a nucleotide triphosphate-binding motif. We have mapped the gene locus of the murine Orc1 gene to chromosome 4, band D. Conservation of structures among members of the ORC1-related proteins suggests that these proteins play a key role in the intiation of DNA replication in all eukaryotes. PMID- 9862485 TI - Promoter trapping identifies real genes in C. elegans. AB - Promoter trapping involved screening uncharacterized fragments of C. elegans genomic DNA for C. elegans promoter activity. By sequencing the ends of these DNA fragments and locating their genomic origin using the available genome sequence data, promoter trapping has now been shown to identify real promoters of real genes, exactly as anticipated. Developmental expression patterns have thereby been linked to gene sequence, allowing further inferences on gene function to be drawn. Some expression patterns generated by promoter trapping include subcellular details. Localization to the surface of particular cells or even particular aspects of the cell surface was found to be consistent with the genes, now associated with these patterns, encoding membrane-spanning proteins. Data on gene expression patterns are easier to generate and characterize than mutant phenotypes and may provide the best means of interpreting the large quantity of sequence data currently being generated in genome projects. PMID- 9862486 TI - Isolation and characterization of a new transposable element in Chlamydomonas reinhardtii. AB - A new transposable element, Tcr3, was identified in the unicellular green alga Chlamydomonas reinhardtii. The Tcr3 element contained imperfect terminal inverted repeat sequences of 56 bp and created a 2 bp target site duplication upon insertion. Insertion of Tcr3 into the 3'-untranslated region of the NIT8 gene, which is essential for nitrate assimilation, prevented expression of the gene. Excision of the Tcr3 element correlated with reversion of the mutant phenotype and left behind a 3 bp footprint. Tcr3 was found in all Chlamydomonas isolates tested and should prove to be useful for transposon-tagging experiments in Chlamydomonas. PMID- 9862487 TI - Nucleotide sequences and functional characterization of two tobacco UAG suppressor tRNA(Gln) isoacceptors and their genes. AB - We isolated and sequenced the two major tRNA(Gln) isoacceptors with CUG and UmUG anticodons from the cytoplasm of Nicotiana rustica. These are the first tRNAs(Gln) of nuclear origin characterized in plants. The tRNA(Gln) sequences were used to design probes for the isolation of the corresponding genes from a nuclear DNA library of N. rustica. The two cloned Nicotiana tRNA(Gln) genes, coding for either of the two isoacceptors, are efficiently transcribed in HeLa cell nuclear extract. In vitro translation in the presence of purified Nicotiana tRNAs(Gln) was carried out in a wheat germ extract partially depleted of endogenous tRNAs. Cytoplasmic (cyt) tRNA(Gln)CUG and to a lesser extent cyt tRNA(Gln)UmUG stimulated readthrough over the UAG stop codon present in the tobacco mosaic virus-specific context. The two tRNA(Gln) isoacceptors are the second class of natural UAG suppressors identified in plants, in addition to cyt tRNA(Tyr)GpsiA which has previously been characterized as the first natural UAG suppressor. PMID- 9862488 TI - Prediction of functional regions of the maize streak virus replication-associated proteins by protein-protein interaction analysis. AB - The replication of the geminiviruses depends on the viral encoded early (complementary-sense) proteins and on host genome encoded factors. Additionally, some of the early proteins (the AL2 protein of subgroup III, and the RepA (formerly known as C1) or Rep (C1:C2) proteins of subgroup I geminiviruses) can function as transcriptional activators of virion- (V-)sense gene expression. The yeast two-hybrid system has allowed us to predict some of the functionally important regions of the maize streak virus (MSV) early proteins RepA and Rep. Defined domains of these proteins were shown to act as transactivators in yeast cells. We detected the association of the RepA and Rep proteins, and their subfragments, with the maize retinoblastoma protein (ZmRb1) which is likely to be one of the interacting host proteins. We showed the self-association capability of the MSV proteins and suggest that homo- or hetero-oligomerization may play an important role in virus replication. These results provide new insights into the role of different regions of the MSV proteins in relation to transcriptional activation and regulation of viral DNA replication. PMID- 9862490 TI - The presence of CYP79 homologues in glucosinolate-producing plants shows evolutionary conservation of the enzymes in the conversion of amino acid to aldoxime in the biosynthesis of cyanogenic glucosides and glucosinolates. AB - A cDNA encoding CYP79B1 has been isolated from Sinapis alba. CYP79B1 from S. alba shows 54% sequence identity and 73% similarity to sorghum CYP79A1 and 95% sequence identity to the Arabidopsis T42902, assigned CYP79B2. The high identity and similarity to sorghum CYP79A1, which catalyses the conversion of tyrosine to p-hydroxyphenylacetaldoxime in the biosynthesis of the cyanogenic glucoside dhurrin, suggests that CYP79B1 similarly catalyses the conversion of amino acid(s) to aldoxime(s) in the biosynthesis of glucosinolates. Within the highly conserved 'PERF' and the heme-binding region of A-type cytochromes, the CYP79 family has unique substitutions that define the family-specific consensus sequences of FXP(E/D)RH and SFSTG(K/R)RGC(A/I)A, respectively. Sequence analysis of PCR products generated with CYP79B subfamily-specific primers identified CYP79B homologues in Tropaeolum majus, Carica papaya, Arabidopsis, Brassica napus and S. alba. The five glucosinolate-producing plants identified a CYP79B amino acid consensus sequence KPERHLNECSEVTLTENDLRFISFSTGKRGC. The unique substitutions in the 'PERF' and the heme-binding domain and the high sequence identity and similarity of CYP79B1, CYP79B2 and CYP79A1, together with the isolation of CYP79B homologues in the distantly related Tropaeolaceae, Caricaceae and Brassicaceae within the Capparales order, show that the initial part of the biosynthetic pathway of glucosinolates and cyanogenic glucosides is catalysed by evolutionarily conserved cytochromes P450. This confirms that the appearance of glucosinolates in Capparales is based on a cyanogen 'predisposition'. Identification of CYP79 homologues in glucosinolate-producing plants provides an important tool for tissue-specific regulation of the level of glucosinolates to improve nutritional value and pest resistance. PMID- 9862489 TI - Interaction of DNA-binding proteins with the 5'-flanking region of a cytokinin responsive cucumber hydroxypyruvate reductase gene. AB - Transcription of the cucumber hpr-A gene is responsive to cytokinin and light. To investigate the molecular basis for transcriptional regulation by cytokinin, we have identified DNA sequences and proteins that may be involved in the regulation of hpr-A gene expression. Transient expression assays in etiolated cucumber cotyledons indicate that the 315 bp fragment (-382 to -67) contains sequences necessary for cytokinin responsiveness of the luciferase reporter gene. Band shift assays detected cytokinin-enhanced and -reduced protein binding sites in a 97 bp fragment (-382 to -285) upstream of the hpr-A gene. DNase I footprinting identified two protein-protected sites, a 15 bp sequence, 5'-AAATGACGAAAATGC-3', that contains an as-1 TGACG motif found in other plant promoters, and a 13 bp sequence, 5'-AAGATTGATTGAG-3', of unknown function. Two-dimensional band shift analysis of the cytokinin-responsive DNA protein complex revealed the presence of six DNA protein interactions. Band shift assays showed that cytokinin and light have different effects on the interaction of nuclear proteins to the 97 bp fragment of the hpr-A gene. These data suggest that cytokinin and light do not share identical signal transduction pathways in regulating hpr-A gene expression. PMID- 9862491 TI - Cortical tissue-specific accumulation of the root-specific ns-LTP transcripts in the bean (Phaseolus vulgaris) seedlings. AB - The characterization of a cDNA clone encoding non-specific lipid transfer protein (PvLTP, formerly named PVR3) in the roots of bean seedlings has been previously reported. In this study, we examined the temporal and spatial accumulation of PvLTP mRNA and the effect of the auxin naphthaleneacetic acid (NAA) on the accumulation of PvLTP mRNA during root development. In situ hybridization showed that accumulation of PvLTP mRNA is highly tissue-specific. Accumulation was detected in the cortical tissue, but not in other tissues of root, including the quiescent center and root cap. Within the cortical tissue, accumulation of PvLTP mRNA was developmentally regulated; accumulation of PvLTP mRNA was high in the cortical tissue of the proximal and ground meristem and declined as cortical tissue developed further. Since the appropriate distribution of auxin is an important factor responsible for the maintenance of root meristem organization. We examined effect of auxin on the accumulation of PvLTP mRNA in relation to the development of cortical tissue. In bean seedlings grown on medium supplemented with 5 microM NAA, morphological alternations, including radial root expansion and abnormal tissue organization in the root apical meristem, were observed. Only faint accumulation signals of PvLTP mRNA were observed in the cortical tissue of proximal meristem region, indicating that cortical tissue development was repressed by exogenous NAA. However, our results suggest that the change in accumulation of PvLTP mRNA is not direct regulatory effect but reflective effect of altered development of cortical tissue that was induced by exogenous NAA. The temporal and spatial accumulation of PvLTP mRNA indicates that PvLTP is a useful marker for the development of cortical tissue in the root tip in bean seedlings. PMID- 9862492 TI - A 42 bp fragment of the pmas1' promoter containing an ocs-like element confers a developmental, wound- and chemically inducible expression pattern. AB - Synthesis of mannopine in plant tissues infected with Agrobacterium tumefaciens is controlled by a divergent promoter (pmas2' and pmas1') that in 479 bp contains all the cis-acting elements necessary to direct tissue-specific and wound inducible expression. In this report, using transgenic tobacco plants harboring a pmas1'-beta-glucuronidase (GUS) gene fusion, we investigated the developmental expression pattern directed by pmas1' in the early stages of development and the responses of pmas1' to different chemical inducers. It was found that this promoter can respond to auxins, cytokinins, methyl jasmonate (MJ), salicylic acid (SA) and its analogue 2,6-dichloroisonicotinic acid (iNA). Treatment with chemical inducers also showed that the effects of iNA are organ-dependent, that wound-induction is a complex response mediated by at least two different chemical signals, and that MJ stimulates changes in the tissue-specific and developmental expression pattern directed by the ptmas1' promoter. Using chimeric promoters we demonstrate that an ocs-like element (ocs+1) directs MJ responses in an orientation-dependent manner and that sequences around the ocs+1 are important to maintain the inducible and developmental properties of this cis-regulatory element. PMID- 9862493 TI - Isolation and characterization of two different cDNAs of delta1-pyrroline-5 carboxylate synthase in alfalfa, transcriptionally induced upon salt stress. AB - Two different cDNA clones, MsP5CS-1 and MsP5CS-2, encoding delta1 -pyrroline-5 carboxylate synthase (P5CS). the first enzyme of the proline biosynthetic pathway, were isolated from a lambdaZap-cDNA library constructed from salt stressed Medicago sativa roots. MsP5CS-1 (2.6 kb) has an open reading frame of 717 amino acids, as well as a non-spliced intron at a position corresponding to the evolutionary fusion point of the bacterial proA and proB genes. MsP5CS-2 (1.25 kb) is a partial clone. The clones share 65% identity in nucleotide sequences, 74% homology in deduced amino acid sequences, and both show a high similarity to Vigna aconitifolia and Arabidopsis thaliana P5CS cDNA clones. Southern blot analysis confirmed the presence of two different P5CS genes. The effect of salinity on the transcription of MsP5CS-1 and MsP5CS-2 in roots was studied, using northern blot analysis and a RT-PCR approach. A rapid increase in the steady-state transcript level of both genes in roots was observed by RT-PCR upon exposure of hydroponically grown 6-day old seedlings to 90 mM NaCl, suggesting that both are salt-inducible genes, yet a higher response was observed for MsP5CS-2. PMID- 9862494 TI - Expression of two consecutive genes of a secondary metabolic pathway in transgenic tobacco: molecular diversity influences levels of expression and product accumulation. AB - We have created a population of transgenic tobacco plants carrying cDNAs encoding two consecutive enzymes from early stages in monoterpenoid alkaloid biosynthesis in Catharanthus roseus. The cDNAs, encoding tryptophan decarboxylase (tdc) and strictosidine synthase (str1) together with a selectable marker gene, were introduced on a single transforming plasmid into tobacco leaves by particle bombardment. Analysis of 150 independent transgenic plants at the DNA and RNA levels demonstrated a range of integration events and steady-state transcript levels for the tdc and str1 transgenes. Southern blot analysis indicated that the tdc and str1 transgenes were integrated at least once in all 150 transformants giving a 100% co-integration frequency of the two unselected genes carried on the same plasmid. A comparison of Southern and northern data suggested that in 26% of the plants, both tdc and str1 transgenes were silenced, 41% demonstrated a preferential silencing of either the tdc or the str1 transgene, with the remaining 33% of the plants expressing both transgenes. We observed no clear correlation between the number of integration events of a specific transgene and the levels of accumulated transcript. Twenty plants representing the range of molecular diversity in the transgenic population were selected for further analysis. Seeds were collected from self-fertilised transformants and germinated on medium containing kanamycin. Seedlings were harvested after 7 weeks and TDC and STR1 enzymatic assays were carried out. We observed a 24- and 110-fold variation in levels of TDC and STR1 activities, respectively. Our data correlate molecular diversity with biochemistry and accumulation of end-product and provide a detailed molecular and biochemical characterization of transgenic plants transformed with a single plasmid carrying two genes of secondary metabolism. PMID- 9862495 TI - A novel protein associated with citrus blight has sequence similarities to expansin. AB - A protein associated with citrus blight (CB), a disease of unknown cause, was partially characterized. The 12 kDa protein, designated p12, is diagnostic of CB and is present in leaves and xylem fluid from roots and stems of CB-affected trees. The protein, and up to six other CB-specific proteins, are readily detected by SDS-PAGE of xylem fluid from CB-affected trees. The partial N terminal amino acid sequence of p12 was found to be unique based on database searches. A cDNA library from CB-affected root cambium was screened with a 60 bp fragment, obtained by PCR amplification of cDNA with degenerate primers designed using the amino acid sequence of p12, and two clones were selected. These clones were sequenced revealing a 674 nucleotide cDNA with a 393 nt ORF which included sequence predicted by the N-terminal amino acid sequence of p12. The amino acid sequence based on the p12 ORF was found to be up to 49% similar and 31% identical to expansins. Bacterial expression of the cloned ORF, which encodes an 11.8 kDa protein plus an N-terminal hydrophobic signal peptide, produced an immunoreactive protein of the expected size. By northern blot analysis, it was determined that p12 transcripts are present in root and stem cambium, but not in leaves of CB affected trees, suggesting transport of the protein to leaves. Southern hybridization analysis of citrus genomic DNA indicated that p12 is a citrus encoded protein. PMID- 9862496 TI - Ethylene-responsive element binding protein (EREBP) expression and the transcriptional regulation of class I beta-1,3-glucanase during tobacco seed germination. AB - Class I beta-1,3-glucanase (betaGLU I) is transcriptionally induced in the micropylar endosperm just before its rupture prior to the germination (i.e. radicle emergence) of Nicotiana tabacum L. cv. 'Havana 425' seeds. Ethylene is involved in endosperm rupture and high-level betaGLU I expression; but, it does not affect the spatial and temporal pattern of betaGLU I expression. A promoter deletion analysis of the tobacco betaGLU I B gene suggests that (1) the distal - 1452 to - 1193 region, which contains the positively acting ethylene-responsive element (ERE), is required for high-level, ethylene-sensitive expression, (2) the regions - 1452 to - 1193 and -402 to 0 contribute to downregulation by abscisic acid (ABA), and (3) the region -402 to -211 is necessary and sufficient for low level micropylar-endosperm-specific expression. Transcripts of the ERE-binding proteins (EREBPs) showed a novel pattern of expression during seed germination: light or gibberellin was required for EREBP-3 and EREBP-4 expression; EREBP-4 expression was constitutive and unaffected by ABA or ethylene; EREBP-3 showed transient induction just before endosperm rupture, which was earlier in ethylene treated seeds and inhibited by ABA. No expression of EREBP- and EREBP-2 was detected. In contrast to betaGLU I, EREBP-3 and EREBP-4 were not expressed specifically in the micropylar endosperm. The results suggest that transcriptional regulation of betaGLU I could depend on: activation of ethylene signalling pathways acting via EREBP-3 with the ERE as the target, and ethylene independent signalling pathways with targets in the proximal promoter region that are likely to determine spatial and temporal patterns of expression. PMID- 9862497 TI - Molecular characterization of tobacco ribonucleotide reductase RNR1 and RNR2 cDNAs and cell cycle-regulated expression in synchronized plant cells. AB - Eukaryotic ribonucleotide reductase (RNR), the enzyme involved in the synthesis of the deoxyribonucleotides, consists of two R1 and R2 subunits whose activities and gene expression are differentially regulated during the cell cycle and are preferentially induced at the G1/S transition. We have isolated three cDNA clones from a tobacco S phase library, two encoding the large R1 subunit, the first cloned in plants, and one encoding the small R2 subunit. From Southern blot hybridization we deduce that RNR2 is encoded by a single-copy gene whereas RNR1 is encoded by a small multigene family. The level of RNR mRNA is cell-cycle regulated showing a maximum in S phase. In mid-S phase, RNR2 transcripts show a higher maximum level than RNR1 transcripts. Analysis of the effects of various cell cycle inhibitors added to freshly subcultured stationary phase cells leads to the conclusion that RNR gene induction at the entry of the cells into the cell cycle takes place in late G1-early S phase. Addition of DNA synthesis-blocking agents to cycling cells synchronized in mid-S phase resulted in an enhancement of RNR transcript level, thus suggesting that RNR gene expression may be linked to the DNA synthesis rate by a feedback-like regulatory mechanism. PMID- 9862498 TI - Isolation and characterization of an Arabidopsis thaliana C-8,7 sterol isomerase: functional and structural similarities to mammalian C-8,7 sterol isomerase/emopamil-binding protein. AB - The yeast C-8,7 sterol isomerase contains a polyvalent high-affinity drug binding site similar to mammalian sigma receptors. Exogenously supplied sigma ligands inhibit sterol biosynthesis in yeast, demonstrating a pharmacological relationship between sigma ligand-binding and C-8,7 sterol isomerase activity. We report the isolation of an Arabidopsis thaliana C-8,7 sterol isomerase by functional complementation of the corresponding sterol mutant in yeast and its characterization by exposure to sigma ligands. The yeast erg2 mutant, which lacks the C-8,7 sterol isomerase gene and activity, was transformed with an Arabidopsis cDNA yeast expression library. Transformed colonies were selected for restoration of C-8,7 sterol isomerase activity (i.e. wild-type ergosterol production) by enhanced resistance to the antibiotic cycloheximide. Sterols produced in complemented lines were characterized by gas chromatography-mass spectroscopy (GC MS). The full-length A. thaliana cDNA (pA.t.SI1) that complemented the erg2 mutation contains an open reading frame encoding a 21 kDa protein that shares 68% similarity and 35% amino acid identity to the recently isolated mouse C-8,7 sterol isomerase. The sigma ligands, haloperidol, ifenprodil and verapamil inhibited the production of ergosterol in wild-type Saccharomyces cerevisiae and in the erg2 mutant complemented with pA.t.SI1. Structural and biochemical similarities between the A. thaliana C-8,7 sterol isomerase and the mammalian emopamil-binding protein (EBP) are discussed. PMID- 9862499 TI - DNase1 footprints suggest the involvement of at least three types of transcription factors in the regulation of alpha-Amy2/A by gibberellin. AB - To elucidate the mechanisms by which alpha-amylase genes are expressed in wild oat aleurone, two genes, alpha-Amy2/A and alpha-Amy2/D, were isolated. Both were shown to be positively regulated by gibberellin (GA) during germination and both contain the conserved cis-acting elements Box 2, GA-response element (TAACAGA) and TATCSATSS (where S is C or G). In addition, they possess a conserved initiator element (CATCA) that is present in both alpha-Amy2 and alpha-Amy1 genes, and also in a number of other plant TATA-containing and TATA-less promoters. DNase 1 footprint analysis showed the alpha-Amy2/A promoter to be a complex array of binding sites for a number of different classes of DNA-binding proteins. Our data suggest that the area around the initiator element (Inr) is bound by a large complex of general transcription factors, that the TATA box is bound by the TFIID complex, that Box 2 is bound by one or more WRKY proteins and that the GA-response element is bound by one or more MYBs. Two other elements containing the core sequence CCATGG/C are bound by nuclear protein and this sequence is the core of the Sph element. The regulation of alpha-Amy2 genes by GA therefore involves an interplay of at least three different types of transcription factor. PMID- 9862500 TI - Arabidopsis cDNA encoding a membrane-associated protein with an acyl-CoA binding domain. AB - Acyl-CoA binding proteins (ACBPs) are small (ca. 10 kDa) highly-conserved cytosolic proteins that bind long-chain acyl-CoAs. A novel cDNA encoding ACBP1, a predicted membrane protein of 24.1 kDa with an acyl-CoA binding protein domain at its carboxy terminus, was cloned from Arabidopsis thaliana. At this domain, ACBP1 showed 47% amino acid identity to Brassica ACBP and 35% to 40% amino acid identity to yeast, Drosophila, bovine and human ACBPs. Recombinant (His)6-ACBP1 fusion protein was expressed in Escherichia coli and was shown to bind 14[C]oleoyl-CoA. A hydrophobic domain, absent in the 10 kDa ACBPs, was located at the amino terminus of ACBP1. Using antipeptide polyclonal antibodies in western blot analysis, ACBP1 was shown to be a membrane-associated glycosylated protein with an apparent molecular mass of 33 kDa. The ACBP1 protein was also shown to accumulate predominantly in siliques and was localized to the seed within the silique. These results suggest that the biological role of ACBP1 is related to lipid metabolism in the seed, presumably in which acyl-CoA esters are involved. Northern blot analysis showed that the 1.4 kb ACBP1 mRNA was expressed in silique, root, stem, leaf and flower. Results from Southern blot analysis of genomic DNA suggest the presence of at least two genes encoding ACBPs in Arabidopsis. PMID- 9862501 TI - Isolation and characterization of a mutant protoporphyrinogen oxidase gene from Chlamydomonas reinhardtii conferring resistance to porphyric herbicides. AB - In plant and algal cells, inhibition of the enzyme protoporphyrinogen oxidase (Protox) by the N-phenyl heterocyclic herbicide S-23142 causes massive protoporphyrin IX accumulation, resulting in membrane deterioration and cell lethality in the light. We have identified a 40.4 kb genomic fragment encoding S 23142 resistance by using transformation to screen an indexed cosmid library made from nuclear DNA of the dominant rs-3 mutant of Chlamydomonas reinhardtii. A 10.0 kb HindIII subclone (Hind10) of this insert yields a high frequency of herbicide resistant transformants, consistent with frequent non-homologous integration of the complete RS-3 gene. A 3.4 kb XhoI subfragment (Xho3.4) yields rare herbicide resistant transformants, suggestive of homologous integration of a portion of the coding sequence containing the mutation. Molecular and genetic analysis of the transformants localized the rs-3 mutation conferring S-23142 resistance to the Xho3.4 fragment, which was found to contain five putative exons encoding a protein with identity to the C-terminus of the A rabidopsis Protox enzyme. A cDNA clone containing a 1698 bp ORF that encodes a 563 amino acid peptide with 51% and 53% identity to Arabidopsis and tobacco Protox I, respectively, was isolated from a wild-type C. reinhardtii library. Comparison of the wild-type cDNA sequence with the putative exon sequences present in the mutant Xho3.4 fragment revealed a G-->A change at 291 in the first putative exon, resulting in a Val-->Met substitution at a conserved position equivalent to Val-389 of the wild-type C. reinhardtii cDNA. A sequence comparison of genomic Hind10 fragments from C. reinhardtii rs-3 and its wild-type progenitor CC-407 showed this G-->A change at the equivalent position (5751) within exon 10. PMID- 9862502 TI - Fusions between green fluorescent protein and beta-glucuronidase as sensitive and vital bifunctional reporters in plants. AB - By fusing the genes encoding green fluorescent protein (GFP) and beta glucuronidase (GUS) we have created a set of bifunctional reporter constructs which are optimized for use in transient and stable expression studies in plants. This approach makes it possible to combine the advantage of GUS, its high sensitivity in histochemical staining, with the advantages of GFP as a vital marker. The fusion proteins were functional in transient expression studies in tobacco using either DNA bombardment or potato virus X as a vector, and in stably transformed Arabidopsis thaliana and Lotus japonicus plants. The results show that high level of expression does not interfere with efficient stable transformation in A. thaliana and L. japonicus. Using confocal laser scanning microscopy we show that the fusion constructs are very suitable for promoter expression studies in all organs of living plants, including root nodules. The use of these reporter constructs in the model legume L. japonicus offers exciting new possibilities for the study of the root nodulation process. PMID- 9862503 TI - Isolation of a cDNA for tryptophan synthase beta from rice and studies of its expression in a 5-methyltryptophan-resistant mutant of rice. AB - A cDNA clone encoding rice tryptophan synthase beta (TSB) was isolated and its transcript level was examined in TR-No. 73, a derivative of a previously isolated rice mutant (TR-1) that is resistant to 5-methyltryptophan. The cDNA sequence of the rice gene for TSB was very similar to that of genes for TSB from other plants. Northern blotting analysis revealed that the steady-state level of TSB mRNA in the 5MT-resistant mutant TR-No. 73 was 1.3 times higher than the level of TSB mRNA in control rice plants under standard conditions. The level of TSB mRNA in control rice plants decreased after treatment of plants with 5MT. Similarly, the level of TSB mRNA in TR-No. 73 initially decreased, although less so than in control rice. However, after 12 h of treatment with 5MT, the level of the transcript in TR-No. 73 returned to the level under standard conditions. The activity of tryptophan synthase (TS) in seedlings of TR-No. 73 was about 2.3 times higher than that in seedlings of control rice under standard conditions. PMID- 9862504 TI - Molecular cloning in Arabidopsis thaliana of a new protein phosphatase 2C (PP2C) with homology to ABI1 and ABI2. AB - We report the cloning of both the cDNA and the corresponding genomic sequence of a new PP2C from Arabidopsis thaliana, named AtP2C-HA (for homology to ABI1/ABI2). The AtP2C-HA cDNA contains an open reading frame of 1536 bp and encodes a putative protein of 511 amino acids with a predicted molecular mass of 55.7 kDa. The AtP2C-HA protein is composed of two domains, a C-terminal PP2C catalytic domain and a N-terminal extension of ca. 180 amino acid residues. The deduced amino acid sequence is 55% and 54% identical to ABI1 and ABI2, respectively. Comparison of the genomic structure of the ABI1, ABI2 and AtP2C-HA genes suggests that they belong to a multigene family. The expression of the AtP2C-HA gene is up regulated by abscisic acid (ABA) treatment. PMID- 9862505 TI - Tissue-specific expression of genes encoding isoforms of the mitochondrial ATPase beta subunit in Nicotiana sylvestris. AB - We report here that the catalytic beta subunit of the mitochondrial ATPase/ATP synthase is encoded by a small multigenic family in the diploid tobacco Nicotiana sylvestris (nsatp2 genes). cDNAs and genes corresponding to the beta1, beta2 and beta3 (pollen specific) isoforms previously detected by 2D-SDS PAGE were isolated. Nsatp2.1 and nsatp2.2 transcripts were found in all vegetative and reproductive tissues analysed. In contrast, nsatp2.3 transcripts were found exclusively in bicellular pollen. As a whole, steady-state transcript levels of nuclear nsatp2 and mitochondrial atp1 genes were found to be closely correlated. PMID- 9862508 TI - Role of p53 mutations in the radiosensitivity status of tumor cells. AB - Wild-type p53 is involved in cellular response to DNA damage including cell cycle control, DNA repair and activation of apoptosis. Accumulation of p53 protein following DNA damage may initiate the apoptotic process, resulting in cell death. DNA damage induced by radiation is an example of apoptotic stimulus involving p53. Regulation of apoptosis by p53 can occur through transcriptional regulation of pro-apoptotic (e.g. bax) and anti-apoptotic (e.g. bel-2) factors. Although wild-type p53 usually sensitizes cells to radiation therapy, p53 mutations have a variable effect on radiation response. For example p53 mutations in bone or breast tumors have been found to be associated with resistance to chemotherapeutic drugs or ionizing radiation. Mutated p53 has has been reported to increase sensitivity to radiation and drugs in colorectal and bladder tumors. The present brief commentary tries to find an explanation at molecular level of these conflicting results. PMID- 9862507 TI - Cloning, sequence and characterization of a sunflower (Helianthus annuus L.) pathogen-induced gene showing sequence homology with auxin-induced genes from plants. AB - The establishment of a plant-pathogen interaction involves changes in gene expressions in both organisms. To isolate Helianthus annuus genes whose expression is induced during processes of resistance to Plasmopara halstedii, a comparison of the expression pattern of healthy sunflowers was made with sunflowers infected with 2 races of P. halstedii, either virulent or avirulent, using differential display of mRNA. A full-length cDNA, HaAC1, representing a sunflower gene whose expression is enhanced during early stages of the incompatible interaction, was isolated. Different timing of RNA accumulation is observed between compatible and incompatible combinations. Sequence analysis and database search revealed significant homology with auxin-induced genes from plants. The expression of this gene, is also induced after treatment with 2,4 dichlorophenoxyacetic acid (2,4-D), salicylic acid (SA) and wounding. PMID- 9862509 TI - Benign breast surgical biopsies: are they always justified? AB - AIM OF THE STUDY: To evaluate the indications for open surgical biopsy of breast lesions resulting in a benign histologic report. METHODS: A consecutive series of 754 benign breast biopsies was collected from six Italian centers previously participating in a multicenter study on the benign/malignant biopsy ratio. Histologic diagnosis, diagnostic tests performed, final clinical diagnosis and the indication for surgical biopsy were compared. RESULTS: Fibrocystic alterations represented the most frequent histologic type (43.2%), followed by fibroadenomas (34.5%). Atypical hyperplasia, phyllode tumors and cancer-like lesions (radial scar, sclerosing adenosis) accounted for a minority of cases. The diagnostic approach was different among centers, with mammography, ultrasonography or cytology being underused in some of them. Suspicion of cancer was an indication for surgical biopsy in 66.7% of cases. In the remaining cases the final report was negative or benign, but biopsy was advised for growing lesions (11.3%) or for cosmetic (3%) or psychological reasons (8.2%). In 4% of cases surgical biopsy was presumably advised for the concurrent influence of high risk conditions such as previous breast cancer (0.7%), family history of breast cancer (2%) or contralateral synchronous breast cancer (1.3%). In 6.8% of the cases biopsy was advised elsewhere for unknown reasons. The indications for biopsy differed among centers, with one center having a low rate of suspicious cases (37%) and a high rate of reported "cosmetic" or "psychological" reasons (47%). CONCLUSIONS: Leaving aside differences in diagnostic approach and aggressiveness, two thirds of all lesions were biopsied in order to exclude cancer. The routine use of a more complete diagnostic protocol and/or alternative methods to obtain a histologic diagnosis (e.g. core biopsy) might substantially reduce the need for open surgical biopsy. PMID- 9862506 TI - Chimeric calcium/calmodulin-dependent protein kinase in tobacco: differential regulation by calmodulin isoforms. AB - cDNA clones of chimeric Ca2+/calmodulin-dependent protein kinase (CCaMK) from tobacco (TCCaMK-1 and TCCaMK-2) were isolated and characterized. The polypeptides encoded by TCCaMK-1 and TCCaMK-2 have 15 different amino acid substitutions, yet they both contain a total of 517 amino acids. Northern analysis revealed that CCaMK is expressed in a stage-specific manner during anther development. Messenger RNA was detected when tobacco bud sizes were between 0.5 cm and 1.0 cm. The appearance of mRNA coincided with meiosis and became undetectable at later stages of anther development. The reverse polymerase chain reaction (RT-PCR) amplification assay using isoform-specific primers showed that both of the CCaMK mRNAs were expressed in anther with similar expression patterns. The CCaMK protein expressed in Escherichia coli showed Ca2+-dependent autophosphorylation and Ca2+/calmodulin-dependent substrate phosphorylation. Calmodulin isoforms (PCM1 and PCM6) had differential effects on the regulation of autophosphorylation and substrate phosphorylation of tobacco CCaMK, but not lily CCaMK. The evolutionary tree of plant serine/threonine protein kinases revealed that calmodulin-dependent kinases form one subgroup that is distinctly different from Ca2+-dependent protein kinases (CDPKs) and other serine/threonine kinases in plants. PMID- 9862510 TI - Does long-term exposure to gel-filled silicone implants increase the risk of relapse after breast cancer? AB - BACKGROUND: An increased risk of cancer and autoimmune diseases associated with gel-filled silicone implants, debated by FDA experts since 1991, has given rise to a profusion of literature on the subject. However, such effects have not been adequately investigated in patients with breast cancer. In a previous report we compared 146 breast cancer patients with gel-filled silicone implants for breast reconstruction to 146 control patients in whom no reconstruction had been performed. The observed results were reassuring, as the evolution of the disease after 10 years was better in the reconstruction group than in the control group. We now report the end results of this study with a median follow-up of 13 years after the breast reconstruction (range, 10-20 years). METHOD: The relative risks of detrimental events were estimated with Cox's Proportional Hazards Model, with stratification according to age at diagnosis. RESULTS: The risks of locoregional recurrences and distant metastasis were significantly lower in the BR group than in the control group. The risks of death, of a second breast cancer and of a second primary cancer at a site other than the breast were not significantly different between the two groups of patients. CONCLUSION: Long-term follow-up of patients exposed to gel-filled silicone implants confirms the absence of detrimental effects after breast cancer. The power of our study is, however, below that required to detect a very slight increase in the risks studied. PMID- 9862511 TI - Treatment of malignant gliomas: a new approach. AB - The aim of this study is to describe the authors' experience with intra-arterial ACNU chemotherapy of malignant gliomas. The prognosis of cerebral malignant gliomas remains poor, whatever traditional therapy is applied. ACNU is a well tolerated nitrosourea with a strong antimitotic effect on neurogenic cells both in vitro and in vivo; this drug has enhanced efficacy when used at high concentrations, particularly as an intraarterial infusion. Seventy-six patients have been studied to date, 68 of whom are evaluable; these patients were treated by intra-arterial infusion of ACNU (100 mg/m2) every 6 weeks, with a mean of 2.5 courses per patient. The objective response (OR) was 28% and analysis of pretreatment factors revealed that survival was influenced by histological grade, other types of therapy applied, and age. In general IAC is well tolerated and the response and survival appear to be better than with systemic chemotherapy. PMID- 9862512 TI - Interleukin-2, interferon-alpha and interleukin-2 plus interferon-alpha in renal cell carcinoma. A randomized phase II trial. AB - BACKGROUND: The purpose of the present study was to investigate the therapeutic effectiveness of interleukin-2 (IL-2) and interferon (IFN), either alone or in combination, in comparable groups of patients affected by advanced renal cell carcinoma (RCC). PATIENTS AND METHODS: In order to limit selection biases, treatment was allocated on a random basis. Patients randomized to IL-2 alone were scheduled to receive eight rlL-2 24-hour i.v. infusion cycles, days 1 to 4, at a daily dose of 18 x 10(6) lU/m2 for a total of 25 weeks. Patients randomized to IFN alone were scheduled to receive rIFN-alpha at a daily dose of 6 x 10(6) IU/m2, days 1, 3 and 5, every week for a total of 52 weeks. Patients randomized to the combination of IFN and IL-2 were given the same drugs at the same daily doses for a total of 24 weeks. Drug dose was modified according to toxicity. RESULTS: Twenty-three percent (95% CI:+/-17.5) of patients treated with IL-2 alone showed an objective response to treatment (9% CR). The corresponding figures in patients treated with IFN alone or IFN plus IL-2 were 9% (95% CI:+/ 11.9) and 9% (95% CI:+/-11.9), respectively. Complete responses were observed only in patients treated with IL-2. The median duration of response in the IL-2 arm was 18 months (range, 9.5-24). The duration of the two responses achieved by IFN alone was seven and nine months, respectively. The corresponding figures in the two patients responding to the combination of IFN with IL-2 were 19 and 27 months, respectively. Total IL-2 dose appeared to be a major predictor of response. Only a minority of patients experienced grade 3-4 toxicity, the incidence being higher in those treated with IL-2 or IL-2 plus IFN. CONCLUSIONS: Neither IFN nor IL-2 or the combination of the two appear to be very active in patients with advanced RCC, even when trial entry was restricted to patients with relatively indolent disease. This stresses the need for the development of new approaches. PMID- 9862513 TI - Myeloprotective effect of early primary granulocyte-colony stimulating factor during six courses of intensified 5-fluorouracil, epirubicin and cyclophosphamide (120FEC) chemotherapy for advanced breast cancer. Cooperative Group of Study and Treatment of Breast Cancer. AB - AIMS AND BACKGROUND: The neutropenia induced by six courses of an intensified FEC regimen is expected to be checked by early primary administration of G-CSF which is stopped eight days before the next chemotherapy course. Less information is available about megakaryocytic and erythroid toxicity over six courses. METHODS AND STUDY DESIGN: Sixty-six consecutive patients with metastatic breast cancer completed six courses of a randomized treatment with two FEC regimens administered every 21 days, in which 600 mg/m2 of cyclophosphamide and 5-FUwas associated with 60 or 120 mg/m2 of epirubicin (60FEC, 35 patients, vs 120FEC, 31 patients). 120FEC was supported by early primary G-CSF (days 4 to 13). Blood counts were obtained seven times during each course. RESULTS: The non-hematologic toxicity over 364 courses was similar in 60FEC and 120FEC. No cumulative hematologic toxicity was observed for white blood cells (WBC) and platelets (PLT), while for hemoglobin (Hb) a somewhat higher cumulative toxicity was observed with 120FEC than with 60FEC. WBC, PLT and Hb grade III-IV toxicity occurred in 40.1% and 45.6% (P=ns), in 23.1% and 0.8% (P <.0001) and in 15.6% and 3.0% (P <.005) of the two regimens, respectively. There were no febrile or hemorrhagic episodes. The epirubicin relative dose intensity delivered was 1.95 in 120FEC with respect to 60FEC. CONCLUSIONS: Our G-CSF schedule permitted to deliver six courses of 120FEC without any clinically relevant side effects. Grade III-IV leukopenia was similar with 120FEC and 60FEC, while grade III-IV thrombocytopenia and anemia occurred more often with 120FEC than with 60FEC. PMID- 9862514 TI - Early gastric cancer: diagnosis, surgical treatment and follow-up of 45 cases. AB - AIMS AND BACKGROUND: The 5-year survival rate of early gastric cancer (EGC) is 85%-100% after "curative" resection, as compared to 20%-30% in advanced gastric cancer (AGC). Because of this relatively high cure rate, the interest in the diagnosis and therapy of EGC has been steadily increasing. The present study, based on 45 EGCs, is aimed at a critical evaluation of the diagnostic procedures and surgical options. METHODS AND RESULTS: Forty-five patients with early gastric cancer (27 men and 18 women; median age, 62 years; range, 28-84) were diagnosed and operated on. They represented 22.5% of all patients with gastric cancer (200) treated in the period July 1987 to January 1998. Forty-one patients were from the northeastern part of Italy. The most frequent symptom was epigastric pain (84%). Barium upper gastrointestinal radiography findings were strongly suggestive of malignancy in 41 cases (91%). Preoperative histopathological diagnosis of adenocarcinoma was performed in 43 cases (95.5%). In two cases (4.5%) severe epithelial dysplasia (associated with ulcer) was the first diagnosis, but the final diagnosis on the basis of the resected specimens was a well differentiated adenocarcinoma. The primary surgical procedure included i) subtotal distal resection (37 cases) with Billroth 11 (33) and Billroth I (4) reconstructions; ii) total gastrectomy (3) for proximal neoplastic extension; iii) proximal gastric resection (2) for cardial cancer; iv) degastro-total gastrectomy (3) for cancer of the stump. Two patients, previously treated with conservative surgery, underwent degastro-total gastrectomy for neoplastic microfocal extension to the margin of resection and for early anastomotic recurrence, respectively. Mural infiltration was limited to the mucosa and submucosa in 27 and 18 cases, respectively. Lymph node metastases were found in three mucosal and five submucosal tumor cases, involving either the first or the second echelon. No operative deaths or postsurgical complications occurred in this series. In the follow-up period (median, 36 months; range, 3-120) four patients died due to other causes; one developed liver metastases, another developed oropharyngeal cancer and two died of biopsy-proven lung cancer without evidence of gastric cancer recurrence. CONCLUSIONS: The clinical presentation of EGC is aspecific. Preoperative endoscopy with biopsy remains the most sensitive diagnostic procedure. For treatment, subtotal distal gastric resection with lymphadenectomy is the "gold standard" but in some instances total gastrectomy may be indicated. Accurate pathological examination establishes the depth of infiltration, as well as the superficial extension of tumors and the lymph node status. Although the prognosis of EGC is favorable, a medium-term follow-up should be planned. PMID- 9862515 TI - Endometrioid carcinoma of the ovary: a retrospective analysis of 106 cases. AB - AIMS AND BACKGROUND: This report retrospectively analyzes 106 cases of endometrioid carcinoma of the ovary treated at the National Cancer Institute of Milan from 1974 through December 1993. In 12 of the 106 cases (11.3%) a synchronous carcinoma of the uterine body was observed. METHODS AND STUDY DESIGN: Only patients who had previously untreated disease were included in the study. Patients with synchronous tumors were staged according to their ovarian cancer and treated according to the stage of that disease. RESULTS: Thirty-nine patients (36.8%) had stage 1, 17 (16.0%) stage 11, 43 (40.6%) stage III, and 7 (6.6%) stage IV disease. Moderately plus poorly differentiated tumors were present in 76 of the 106 cases (71.7%). Considering the 67 patients with advanced disease, residual tumor was absent in 27 cases (40.3%), < or = 2 cm in 17 (25.4%), and > 2 cm in 23 (34.3%) cases. Systematic pelvic and para-aortic lymphadenectomy was performed in 60 patients (56.6%); selective sampling was carried out in 23 cases (21.7%). After surgery, 77 patients underwent various chemotherapy regimens. CONCLUSION: Using univariate analysis, FIGO stage, tumor grade, residual disease after surgery, lymph node status, and platinum in the chemotherapy regimen significantly influenced 5-year survival. However, when all these variables were included in a multivariate analysis only FIGO stage still had a significant impact on survival. Survival analysis also showed a trend towards longer survival in patients with synchronous tumors. PMID- 9862516 TI - Combined epirubicin and interleukin-2 regimen in the treatment of malignant mesothelioma: a multicenter phase II study of the Italian Group on Rare Tumors. AB - The Italian Group on Rare Tumors undertook a phase II study of a combination of epirubicin and interleukin-2 in 21 chemotherapy-naive patients with malignant mesothelioma. All patients had bidimensionally measurable disease at CT scan. Treatment included Intravenous administration of epirubicin at a dose of 110 mg/m2 i.v. on day 1, and interleukin-2 at a dose of 9 MU subcutaneously from day 8 to day 12 and from day 15 to day 19. Cycles were repeated every three weeks, up to six times in the absence of progressive disease. Treatment response was evaluated after two cycles of therapy. Only one patient achieved a partial response, resulting in an overall response rate of 5% (1/21) with a median progression-free and overall survival of 5 and 10 months, respectively. Toxicity was relevant and caused treatment discontinuation in many patients. These results do not support the use of such a combination in the management of malignant mesothelioma. PMID- 9862517 TI - Treatment of advanced colorectal cancer (CRC) in daily practice: results of a survey in two Italian regions, Piemonte and Valle d'Aosta. AB - AIMS AND BACKGROUND: Colorectal cancer (CRC) is one of the most important health problems in Western countries: it is the fourth cancer in terms of incidence and the second cause of cancer death. Surgery is the main therapeutic choice and there is broad consensus on the role of adjuvant chemotherapy (CT) after resection. Unfortunately, 50% of the patients will relapse and die of the disease. Palliative CT based on 5-fluorouracil (5FU) may induce a 9-48% response rate with a median survival of 11.5 months. At present there is no gold standard for CT In advanced CRC and the situation has become more complicated since the advent of new drugs (Raltitrexed, Irinotecan, Oxaliplatin). The aim of this study was the identification of the different approaches to treatment of advanced CRC among the clinicians (oncologists, radiologists, internal medicine specialists, surgeons) who practice CT. METHODS AND STUDY DESIGN: Forty-six clinicians from two Italian Regions (Piemonte and Valle d'Aosta) were interviewed by telephone. RESULTS: 5FU modulated with Lederfolin according to the classic Machover scheme is the main option in daily practice. More sophisticated therapies are reserved to patients with a good performance status (PS) and are prescribed only in the larger centers. The planned therapies usually consist of six courses. Restaging may be performed after three or six courses. A marked difference has been recorded in the evaluation of a situation of no change (NC): 25.5% of the clinicians evaluate stable disease as a positive result. In the event of disease progression or relapse, 35% of the clinicians do not prescribe second-line CT. In case of further treatment, the options are totally subjective. CONCLUSIONS: A national survey on this issue is necessary under the auspices of AIOM (Associazione Italiana Oncologia Medica) and involving oncologists, epidemiologists and statisticians, in order to define the reasons for variations in therapy in advanced CRC and determine the differences between clinicians of different age, specialization and location. This survey could lead to a definition of guidelines for the treatment of advanced CRC. PMID- 9862518 TI - High-grade cervical intraepithelial neoplasia (CIN) in pregnancy: clinicotherapeutic management. AB - An increasing incidence of cervical intraepithelial neoplasia (CIN) among young women has been noticed in recent years. For this reason pregnancy might represent a peculiar opportunity to undergo cytocolposcopic examination for those women who do not take part in a screening program for cervical carcinoma. Diagnosis of CIN during pregnancy poses the question of the management of this disease and particularly of whether it is better to treat the lesion or not during pregnancy. To contribute to the solution of this issue we initiated a study on the management of high-grade CIN in pregnancy. MATERIAL AND METHODS: Five hundred and seventy-one pregnant women underwent cytologic, colposcopic and, when necessary, histologic examination. Those in whom a CIN was discovered in the first four months of gestation underwent laser conization. When the diagnosis of CIN was made after the sixteenth week of gestation, cytocolposcopic monitoring was performed every eighth week during pregnancy and two months after childbirth. Laser conization was performed under colposcopic guidance in the outpatient setting in all cases. All treated patients were submitted to cytologic, colposcopic and, if necessary, histologic examination every third month in the first year after treatment, every sixth month in the second year and yearly from the third year onwards. RESULTS: In 14 (2.4%) of the 571 examined women a CIN III was discovered, 6 of which associated with a human papilloma virus (HPV) infection. Of these, 8 patients, whose diagnosis was made within the sixteenth week of pregnancy, underwent laser conization. In one case a minor hemorrhage occurred during treatment. Two patients reported minor bleeding up to ten days after treatment. No major hemorrhages or cervical stenosis were observed. Histologic examination of the cones confirmed the preoperative diagnosis based on cervical biopsies and the lesion was entirely removed by conization in all cases. Seven of the 8 patients who underwent laser conization during pregnancy had a spontaneous delivery at term. The remaining patient, who had had a previous cesarean section, was again delivered by cesarean section. All treated patients were cured after the first-year follow-up visit. In 6 patients CIN was diagnosed after the sixteenth week of pregnancy. These women underwent cytocolposcopic examination every eighth week during pregnancy and two months after delivery, when the cervical changes associated with gestation had disappeared. Four of these patients showed persistence of CIN at postpartum follow-up and therefore underwent laser conization. In two patients spontaneous regression of the lesion was observed. In no case did progression to invasive carcinoma occur. CONCLUSIONS: Given the increasing incidence of CIN in young women, the beginning of pregnancy may represent a peculiar opportunity for all pregnant women who do not take part in cervical screening programs to undergo a cytocolposcopic examination. In case of a diagnosis of high-grade CIN within the first 16 weeks of pregnancy, a conservative excisional treatment, which does not expose the pregnancy to any risk, should be carried out in order to confirm the intraepithelial localization of the lesion. PMID- 9862519 TI - Limited role of TP53 and TP53-related genes in myxoid liposarcoma. AB - AIMS: Circumstantial evidence suggests that genetic changes may lead to tumor progression within the myxoid liposarcoma tumors (MLTs) carrying non-random chromosomal translocation t(12;16). METHODS: To address this subject an immunophenotypic analysis, applying antibodies against proteins encoded by TP53, MDM2 and CDK4 genes, complemented by molecular analysis of eight suitable cases, was performed on 104 consecutive cases. Chromosomal translocations were assessed either by cytogenetic analysis or by RT-PCR in 9 suitable cases and chimeric transcripts were found in all cases but two pleomorphic liposarcomas. RESULTS: Based on immunophenotyping and tumor site, the case material consisted of three groups. The first one was made up of 92 non-retroperitoneal cases carrying a null p53, mdm2, cdk4 immunophenotype, which remained unchanged over the time of recurrences and along the gamut of histologic subtypes. The second group was represented by five p53+, mdm2-, cdk4- non-retroperitoneal cases, 4 of which were further analysed by PCR-SSCP for p53 mutation. The immunophenotypic profile of these cases, complemented by the molecular findings, supported a role of TP53 in tumor progression in three high-grade MLTs. The third group, consisting of 7 retroperitoneal cases, showed a heterogeneous immunophenotype, sharing immunophenotypic and molecular features with the well-differentiated/evoluted (dedifferentiated) liposarcoma group. CONCLUSIONS: TP53 mutations seem to play a role in tumor progression in a few cases of MLTs (2.8%) showing more aggressive histologic characteristics. The unexpected finding that a number of retroperitoneal LMTs display the immunophenotypic profile of the well differentiated/evoluted (dedifferentiated) liposarcomas, deserves further investigation. PMID- 9862520 TI - Characterization of a murine monoclonal antibody against human esophageal squamous cell carcinoma-associated antigen. AB - A murine monoclonal antibody (MAb) 2G3 of the IgG1 type was raised using the human esophageal squamous cell carcinoma (SCC) cell line TE-2. Immunoblotting with 2G3 indicated that the antigen recognized by 2G3 has a molecular weight of 34 kD. Its activity was evaluated by immunoperoxidase and immunofluorescence on frozen and paraffin sections of various normal tissues, normal and benign tumors as well as various established cell lines. The pattern of reactivity revealed that the antigen recognized by 2G3 was expressed mainly by esophageal SCC. The only exception was represented by malignant breast tumors, where it reacted weakly. Scatchard analysis using 125I-labelled 2G3 showed that TE-2 has approximately 7.5 times more binding sites than the MCF-7 breast cancer cell line. The use of this new MAb is therefore proposed for the histopathological diagnosis of esophageal SCC. PMID- 9862521 TI - ras and c-myc oncoproteins during tumor progression in the uterine cervix. AB - AIMS AND BACKGROUND: Altered oncogenic activity is a feature associated with many malignant and premalignant conditions. Among the many oncogenes, ras and myc are commonly altered in many tumors. This study aims to evaluate the expression of ras and c-myc oncoproteins in a total of 204 cervical tissue samples, including premalignant and malignant lesions as well as apparently normal cervical tissue. METHODS AND STUDY DESIGN: Mouse monoclonal antibodies against the three mammalian ras gene products (c-H-ras, c-K-ras, c-N-ras) and the c-myc protein were used to evaluate oncoprotein expression by immunocytochemistry. RESULTS: None of the samples analyzed displayed immunoreactivity for H-ras and K-ras. Normal cervical epithelium showed minimal immunoreactivity for N-ras with about 33% of the samples expressing the protein. More conspicuous expression in normal tissue was displayed by c-myc, with about 90% of the samples expressing the protein (mean value of cells positive=34%). The immunoreactivity for N-ras increased with increasing histological abnormality from low-grade squamous intraepithelial lesions (SIL) to invasive carcinoma. Increased immunoreactivity for N-ras was evident in the basaloid cells of malignant lesions, with the maximum value of 66% found in poorly differentiated squamous cell carcinoma (PDSCC). The percentage of nuclei positive for c-myc also showed a gradual increase from low-grade SIL onwards, the highest positivity being found in PDSCC, where the mean value was 85%. Statistical analysis revealed a good correlation between the expression of N ras (r=0.8922, P=0.001) and c-myc (r=0.8856, P=0.001) and various histological stages of tumor progression in the cervical epithelium. CONCLUSIONS: These results therefore suggest that c-myc and N-ras oncoproteins are important during tumor progression in the uterine cervix. PMID- 9862522 TI - Expression of cell surface glycoprotein CD44 and integrins in breast cancers among Indian women. AB - Parsis, the sole surviving group of followers of Zoroaster who are settled in Bombay, have a fourfold higher incidence of breast cancer than the general population of Greater Bombay. CD44 expression was studied by immunohistochemistry in breast cancers of 50 non-Parsi and 35 Parsi women, 10 normal breast tissues, 10 proliferative lesions and 49 tissues adjoining a tumor mass. Alpha2 and beta1 integrins could be studied in only 42 malignant cases and five normal tissues. The immunohistochemistry results were correlated with other parameters including tumor grade and size, estrogen and progesterone receptor status, lymph node involvement and mitotic index. CD44 was not expressed in normal areas. Benign areas and tissues adjacent to tumor masses showed increased staining. Both Parsi and non-Parsi women showed significantly high CD44 expression. All Parsi ILCs were strongly positive for CD44. In both groups ER negativity was associated with strong CD44 positivity, while mitotic counts correlated with decreased CD44 expression in Parsis but not in non-Parsis. Alpha2 and beta1 integrins were strongly expressed on the basolateral surface of normal epithelium. However, they were downregulated in tumors. In general breast tumor tissues from Parsi and non Parsi patients did not differ significantly with respect to most parameters. However, among Parsis lymph node involvement and CD44 correlated weakly whereas the mitotic index was inversely correlated with CD44. The reverse was true for non-Parsis. The deviation from the general pattern needs further study based on a large number of samples and appropriate use of splice variants. PMID- 9862523 TI - Littoral cell angioma of the spleen: an additional report of four cases with emphasis on the association with visceral organ cancers. AB - AIMS AND BACKGROUND: Littoral cell angioma (LCA) is an uncommon vascular tumor of the spleen recently described and interpreted as the tumoral counterpart of the normally present littoral cells lining the splenic sinus channels of red pulp. The diagnosis of LCA is suggested by a quite characteristic morphology and confirmed by the demonstration of a hybrid endothelial/histiocytic phenotype. METHODS: Four original and previously unreported cases of LCA are presented. All four splenic vascular tumors were investigated by light microscopy and immunohistochemistry for endothelial and histiocytic markers. RESULTS: All four cases were associated with visceral epithelial malignancies (colorectal adenocarcinoma in two cases, renal and pancreatic adenocarcinoma in one case each). One case was also associated with an intracranial tentorial meningioma. CONCLUSIONS: We consider our findings as a novelty and signal the possible existence of a clinical syndrome. Five of a total of 21 previously reported cases in the literature were also described as being associated with other cancers (non Hodgkin's lymphoma in two cases, two not further specified tumors of the liver and brain, an epithelial ovarian cancer, and a non-small cell lung cancer in one case each). Close follow-up and careful investigation in search of a second visceral neoplasm are strongly recommended in cases of LCA, but further clinical observations and more in-depth genetic and molecular studies are needed before any valid conclusions can be drawn. PMID- 9862524 TI - Second primary renal cell carcinomas following solid tumors. Four case reports and review of the literature. AB - We report multiple malignancies in four patients with metachronous renal adenocarcinomas. The group of patients includes two women treated for primary breast and ovarian cancer and two men with prostate and colon cancer respectively. The occurrence of renal cancer following these primary solid tumors was not associated with previous treatments. The authors review the literature and discuss pathogenic mechanisms underlying renal cell carcinoma. PMID- 9862525 TI - Accessory breast cancer: a case report and review of the Japanese literature. AB - The case of a 31-year-old woman with accessory breast cancer in the left axilla is described. She had noticed a swelling in the left axilla during her three pregnancies. The preoperative diagnosis of accessory breast cancer was made on the basis of ultrasound-guided fine-needle aspiration biopsy (US-FNAB) and clinical history. She was treated by wide local resection and regional lymph node dissection. Although cancer originating from accessory breast tissue has been reported very rarely, knowledge of this disorder may facilitate the correct diagnosis of axillary tumors. US-FNAB is a useful and simple technique for the tissue diagnosis of axillary tumors. PMID- 9862526 TI - Splenic mucinous cystadenocarcinoma arising in heterotopic pancreatic tissue. AB - A mucinous cystadenocarcinoma along with remnants of normal pancreatic tissue was discovered in the spleen of a 21-year-old woman. Extensive clinical, radiological, ultrasound and computed tomography investigation revealed that the patient's pancreas and ovaries were unremarkable. We presume an origin from heterotopic pancreatic tissue for this neoplasm and present a review of the literature in this regard. PMID- 9862527 TI - Accidental overdose of melphalan per os in a 69-year-old woman treated for advanced endometrial carcinoma. PMID- 9862528 TI - A case of metastatic axillary lymph nodes involvement from unknown primary cancer: clinical usefulness of [99mTc]- sestamibi. PMID- 9862529 TI - Archery as a possible rehabilitative activity in the oncological structure: preliminary observations from a feasibility study. PMID- 9862531 TI - Site-specificity of bone mineral density and muscle strength in women: job related physical activity. AB - We proposed that there are significant correlations between muscle strength and bone mineral density in premenopausal women and that these correlations are site specific. To test this hypothesis, we examined the relationships among site specific bone mineral density, physical activity, and muscle strength in a group of 96 healthy premenopausal Caucasian women. Bone mineral density was measured at the lumbar spine and at three sites in the proximal femur (trochanter, femoral neck, and Ward's triangle) with dual-energy x-ray absorptiometry and at the mid radius with single-photon absorptiometry. The muscle strength of hip and spinal muscle groups was determined with a strain gauge isodynamometer, and grip strength was measured with the JAMAR dynamometer. The strength of shoulder girdle muscle groups was evaluated with the use of free weights. Physical activity was determined by surveying the subjects and by using a standardized scale. Data analysis revealed significant linear correlations of muscle strength with bone mineral density at the mid-radius (r = 0.31; P = 0.002) and at the hip (r = 0.26; P = 0.01). Grip strength was significantly correlated with bone mineral density of both the spine and the femur (r = 0.24, r = 0.34; P < 0.05 for both). Back extensor strength correlated with bone mineral density of the hip (Ward's triangle; r = 0.23; P = 0.023). However, there was no significant positive correlation between the strength of the spinal flexor or extensor muscles and the site-related bone mass (lumbar spine). Only one of the three components of the physical activity score (job) positively correlated with vertebral bone mineral density (r = 0.21; P = 0.04). Physical activity negatively correlated with age (r = 0.24; P = 0.02). We conclude that in premenopausal women, the effect of muscle strength on bone mass is more systemic than site-specific. A positive correlation between vertebral bone mass and components of physical activity demonstrates that even job-related physical activity is an important factor in maintaining adequate bone mass. PMID- 9862530 TI - End-stage acute hepatic failure as clinical presentation of liver metastases from breast cancer. AB - A most unusual case of fatal acute hepatic failure as clinical presentation of liver metastases from breast cancer is described. The patient had a four-year history of indolent breast neoplasm and no previously known liver disease. PMID- 9862532 TI - Relationship between bone mineral density of the proximal femur and strength of hip muscles in postmenopausal women. AB - The relationship between muscle strength and bone density is well known, but only a limited number of studies relating muscle strength to bone mineral density of a site-specific limb have been performed. The main purposes of this study were as follows: (1) to assess the relationship between the strength of hip muscles and bone mineral density of the proximal femur in 100 postmenopausal women; (2) to compare strength values of three subgroups, which were formed on the basis of the femoral neck Z-scores. By using an isokinetic dynamometer (Cybex 350), muscle strength tests were performed for hip muscles and bone mineral density measurements were obtained by dual energy x-ray absorptiometry. Statistical analysis demonstrated a moderately positive correlation between bone mineral density of the femoral neck and hip muscle strength (with abductors: r = 0.267, P = 0.008; with adductors: r = 0.276, P = 0.007). A moderate correlation was also found between bone mineral density of Ward's triangle and hip adductors (r = 0.327; P = 0.001). When muscle strength mean values of subgroups were compared with each other, there was no statistically significant differences. These data may suggest that the isokinetic strength of hip muscles may not contribute to the bone mineral density of the proximal femur. PMID- 9862533 TI - Fibrillation potential amplitude after denervation. AB - Fibrillation potentials have been reported to decline in amplitude with time after denervation. The use of maximum fibrillation potential amplitude to determine the relative acuity of axonal loss ("old" v "new/recurrent") has been advocated but with conflicting endorsements as to the appropriate benchmark amplitude, i.e., 100 microV compared with 250 microV. This investigation uses computer simulations to examine the rate of fibrillation potential amplitude decline expected given known values for muscle fiber size atrophy and conduction velocity slowing over time after denervation. Factors that affect the amplitude and potentially lead to erroneous interpretations in the clinical scenario of partially denervated muscle tissue are discussed. The use of fibrillation potential maximum amplitude criteria to determine the age of lesion onset in both totally and partially denervated muscle is fraught with technical and pathophysiological hazards of interpretation and must be considered cautiously, if at all, in clinical practice. PMID- 9862534 TI - Passive and active exercises increase cerebral blood flow velocity in young, healthy individuals. AB - Unlike the well-examined cardiovascular changes during movement stimuli, changes of cerebral hemodynamics and cerebral metabolism in physical exercises have, as yet, rarely been studied. Our objective was to investigate whether there are changes in cerebral hemodynamics and cerebral metabolism caused by active and passive movement stimuli. Response to repetitive active and passive movement stimuli was studied in 14 volunteers (8 females, 6 males; mean age, 35.2+/-8 yr). Each volunteer was subjected to four measurement series while performing a defined active and passive exercise program of the right upper or right lower limb. Measurement series were designed according to Aaslid's "evoked flow test"; exercises were performed for 20 s, followed by a rest of 20 s. This sequence was repeated 10 times in each series. As a measure of cerebral hemodynamics mean and peak blood flow velocity of the middle cerebral artery were recorded by transcranial Doppler sonography (Multidop-X-DWL). In addition, cerebral metabolism was quantified in a subsample by means of oxygenic cytochrome aa3 (respiratory chain enzyme), cerebral oxygen saturation using near infrared spectroscopy (Critikon). As well, noninvasive blood pressure (Penaz method) and expiratory pCO2 were measured. Compared with resting measures, an increase in mean cerebral blood flow velocity of the middle cerebral artery of 3.8% (P = 0.003, paired t test) was observed after active exercises of the right lower limb and 3.5% after active exercises of the right upper limb. Respective changes were 3.4% (P = 0.004) for passive exercises of the lower limb and 4.6% (P = 0.007) for passive exercises of the right upper limb. Peak cerebral blood flow velocity of the middle cerebral artery showed an even more pronounced increase during passive and active exercises in all cases, with values of between 12.2% (P < 0.001) and 13.6% (P < 0.001). Significant increases (1.5-3%) of cytochrome aa3 and cerebral oxygen saturation were observed during active and passive exercises. These studies show that active as well as passive exercises are followed by an increase of cerebral blood flow velocity. We attribute the increase of cerebral hemodynamic and cerebral metabolism to cerebral activation and autoregulative mechanisms. PMID- 9862535 TI - Acute sensory neuronopathy: identified with electrodiagnosis and magnetic resonance imaging. AB - We report on a 13-year-old female with idiopathic acute sensory neuronopathy mimicking a sensory form of Guillain-Barre syndrome, which was identified by using electrodiagnosis and spine magnetic resonance imaging. Motor conduction results were normal, but no sensory nerve action potentials were seen in the four limbs. On magnetic resonance imaging of the whole spine, the diffuse gadolinium enhancement of the dorsal roots in the spinal canal was detected, without evidence of intramedullary lesions. The clinical symptoms and electrodiagnostic findings had persisted for more than 18 months of follow-up. PMID- 9862536 TI - Bladder volume measurement with electrical impedance analysis in spinal cord injured patients. AB - The purpose of the study contained herein was to determine the usefulness of electrical impedance for measurement of bladder volume in spinal cord-injured patients, with an assessment of the relationship between electrical impedance and bladder volume exclusively. The study was performed during urodynamic studies to match the exact bladder volume. Thirteen patients with complete spinal cord injuries were recruited. We used silver-silver chloride compound electrodes composed of one pair of current and amplitude electrodes to minimize the influence of superficial skin impedance. Each compound electrode was attached on the lower abdomen bilaterally after skin cleansing. Constant current (60 kHz-1.0 mA), converted from 9 V of direct current, was applied, and corresponding electrical impedance (omega) was measured at "pre" (before urodynamic empty bladder), "full" (with a urodynamic filled bladder), and "post" (after urodynamic empty bladder) status. Electrical impedance at the full status was definitely lower than that at the pre and post statuses in all subjects, with a statistically significant difference (P < 0. 001). The correlation between electrical impedance and bladder volume was negative (r = -0.7988), and the fact of how much the variation in electrical impedance could be explained by variation in bladder volume was estimated (r2 = 0.6381). From these findings, we have determined that the electrical impedance analysis technique can be an alternative measure of bladder volume indirectly. PMID- 9862538 TI - Comparison of phenol block and botulinus toxin type A in the treatment of spastic foot after stroke: a randomized, double-blind trial. AB - Locally acting treatments for spasticity such as nerve and motor point blocks have the advantage of reducing harmful spasticity in one area, while preserving useful spasticity in another area. This randomized, double-blind study is the first trial that was designed to find out whether botulinus toxin Type A and phenol relieves the signs and symptoms of ankle plantar flexor and foot invertor spasticity after stroke and if either of these methods offers any advantages and disadvantages over the other. Twenty patients who were included in this preliminary study were randomly assigned to receive a single treatment of 400 mouse units of botulinus toxin Type A injected into the calf muscles or to receive a tibial nerve blockade with 3 ml of 5% phenol. A combination of subjective and objective measures were used to assess functional change at baseline and at Weeks 2, 4, 8, and 12. At follow-up, significant improvement (P < 0.05) in the Ashworth score for dorsiflexion was observed in both groups. The change in the Ashworth score for eversion was significant in the group that received botulinus toxin Type A (P < 0.05) but not in the group that received phenol (P > 0.05). When those variables were compared between the two groups, the change in the Ashworth score at Weeks 2 and 4 was significantly better in the group that received botulinus toxin Type A (P < 0.05) but there was not a significant difference between the two groups at Weeks 8 and 12 (P > 0.05). The decrease in clonus duration that was detected by electromyography was significant in both groups at all visits, but the decrease in the group that received botulinus toxin Type A was significantly better at Weeks 2 and 4 (P < 0.05). It is concluded that both motor point injections with botulinus toxin Type A and tibial nerve blockade with phenol are effective in plantar flexor spasticity, but the changes were more significant in the group that received botulinus toxin Type A at Weeks 2 and 4, whereas there was not a significant difference between the two groups at Weeks 8 and 12. Future research should explore the long-term effect of these two treatment modalities. PMID- 9862537 TI - Gait adaptations during walking under visual and cognitive constraints: a study of patients recovering from limb-saving surgery of the lower limb. AB - The objective of this case series study was reautomatization of gait after limb saving surgery for tumors at a laboratory of gait analysis. Twelve patients (9 males and 3 females; mean age, 38 years) who underwent limb-saving surgery of the lower limb at least 1 year previously and ten normal subjects (3 males and 7 females; mean age, 37.5 years) were studied. The main outcome measures were walking speed and stride time duration under normal walking conditions as well as the use of different types of constraints. Patients walked with a lower preferred walking speed than the normal subjects. Patients showed a higher coefficient of variation of stride time in normal walking as well as complex walking compared with normal subjects. During walking with constraints, a significant decrease in stride time was found in patients but not in normal subjects. Although restoration of gait after limb-saving surgery is impressive, it is not complete (lower walking speed) and can break down under conditions of visual and cognitive load. Hence, the application of complex tasks reveals that gait reautomatization is not complete in these patients during a period of 2 to 5 years after surgery. PMID- 9862539 TI - Comparison of discomfort associated with surface and percutaneous intramuscular electrical stimulation for persons with chronic hemiplegia. AB - Neuromuscular stimulation may facilitate motor recovery after stroke or brain injury, reduce shoulder pain associated with hemiplegia, and reduce cerebral spasticity. However, the discomfort of surface neuromuscular stimulation significantly limits the clinical implementation of this modality for persons with hemiplegia. The study contained herein tests the hypothesis that stroke and brain injury survivors with chronic hemiplegia (>6 mo) and intact sensation tolerate percutaneous intramuscular stimulation better than surface stimulation. Four stroke and two traumatic brain injury survivors participated in the study contained within this article. Each subject received three pairs of percutaneous and surface stimulations of the paretic finger extensors. The order of the type of stimulation within each pair was randomly assigned. The stimulation parameters for each type of stimulation were normalized to produce the same torque at the metacarpophalangeal joint. Subjects rated their perceived level of discomfort using a 10-cm visual analog scale and the McGill Pain Questionnaire. A blinded evaluator administered the pain measures. Percutaneous stimulation was associated with significantly lower discomfort as reflected by the visual analog scale (0.74 v 3.3; 95% confidence interval of difference, -3.84, -1.28). The McGill Pain Questionnaire produced similar results with percutaneous stimulation associated with a significantly fewer number of words chosen to describe the discomfort (0.87 v 3.30; 95% confidence interval of difference, -3.50, -1.30) and significantly lower Pain Rating Index (1.47 v 6.27; 95% confidence interval of difference, -7.77, -1.83). Data suggest that percutaneous intramuscular stimulation is significantly better tolerated than surface stimulation and that percutaneous stimulation may enhance patient compliance with neuromuscular stimulation treatments. PMID- 9862540 TI - Tartaric acid-induced cough and the superior laryngeal nerve evoked potential. AB - The purpose of this study was to stimulate the laryngeal cough reflex using a nebulized, mild chemical irritant and to record an associated laryngeal evoked potential from the internal branch of the superior laryngeal nerve. The laryngeal evoked potential was obtained on ten normal subjects from the right internal branch of the superior laryngeal nerve. The electrodiagnostic setup included an active electrode placed just below the hyoid bone with a 4-cm separation and distal reference. A ground electrode was placed between the active and reference electrodes. The receptors and internal branch of the superior laryngeal nerve were stimulated by inhalation of a nebulized 20% solution of tartaric acid and normal saline. The time line was triggered by a pneumatic switch on initial inspiration of the nebulized tartaric acid. The electrodiagnostic settings were set at a sweep speed of 1 ms/division, a gain of 10 to 20 microV/division, and 20 to 2,000 filters. There were 132 variables recorded from the internal branch of the superior laryngeal nerve of the ten subjects. The mean peak distal latency was 1.66+/-0.42 ms with a 1.6 median, 1.6 mode, and 0.17 variance. The duration was 0.41 ms, and amplitude was 5.19+/-2.91 microV. In conclusion, the laryngeal evoked potential, the afferent component of the involuntary cough reflex, can be recorded from the internal branch of the superior laryngeal nerve after inhalation of tartaric acid-induced cough. PMID- 9862542 TI - High lumbar disc: diagnostic and treatment dilemma. AB - The high lumbar disc refers to a disc herniation involving the upper lumbar discs (L2-4). Radiculopathy caused by an upper lumbar disc is occasionally difficult to diagnose secondary to ambiguous presenting symptoms. Once accurately diagnosed, the treatment program is straight-forward but specific for the high nerve root pathology. A series of three cases of high lumbar radiculopathies managed with an aggressive conservative approach are presented. This case series will review the diagnostic and treatment strategies that are useful in the identification and management of a high lumbar radiculopathy. Various pitfalls in the management of these cases are reviewed, all of which may worsen the patient's clinical situation. A high index of suspicion is necessary to elucidate this entity. PMID- 9862541 TI - Physical performance and cardiovascular and metabolic adaptation of elite female wheelchair basketball players in wheelchair ergometry and in competition. AB - Spinal cord injury leads to a pronounced reduction of cardiovascular, pulmonary, and metabolic ability. Physical activity, up to and including high-performance sports, has obtained importance in the course of rehabilitation and the postclinical phase. Thirteen elite female wheelchair basketball players from the German National Basketball Team and 10 female sedentary spinal cord-injured persons were examined in the study. Heart volume was measured by an echocardiography. All subjects underwent a graded exercise test on a wheelchair ergometer. Additionally, heart rate, lactate, and player points were measured during a competitive basketball game in wheelchair basketball players. Cardiac dimensions were larger for spinal cord-injured wheelchair basketball players (620.3 ml; 9.6 ml x kg(-1)) in comparison with spinal cord-injured persons (477.4 ml; 8.2 ml x kg(-1)) but did not exceed the heart volume of untrained nonhandicapped persons. In contrast, athletes with amputations or those having had poliomyelitis reached training-induced cardiac hypertrophy in relation to body mass (713.7 ml; 13.2 ml x kg(-1)), as observed in nonhandicapped athletes. During graded wheelchair ergometry, wheelchair basketball players showed a higher maximal work rate (59.9 v 45.5 W), maximal oxygen consumption (33.7 v 18.3 ml x min(-1) x kg(-1)), and maximal lactate (9.1 v 5.47 mmol x l(-1)) without a difference in maximal heart rate and workload at AT4 than did spinal cord-injured persons. The average heart rate during the wheelchair basketball game was 151 x min(-1), and the lactate concentration was 1.92 mmol x l(-1). Female athletes with a less severe handicap and higher maximal oxygen consumption during the graded exercise test reached a higher game level in the evaluation. During the competitive basketball game, high cardiovascular stress was observed, indicating a fast aerobic metabolism; the anaerobic lactic acid capacity played a subordinate role. Wheelchair basketball is an effective and suitable sport to enhance physical performance and to induce positive physiological adaptations. PMID- 9862543 TI - Neurogenic atrophy of suboccipital muscles after a cervical injury: a case study. AB - This case report describes abnormalities in bilateral rectus capitis posterior minor muscles in one individual with persistent head and neck pain. These findings are muscle atrophy, fatty infiltration on magnetic resonance imaging, and electromyographic abnormalities compatible with denervated muscle. The objective of the study contained herein was to determine if fatty infiltration on magnetic resonance imaging of the rectus capitis posterior minor muscle is the result of disuse or denervation. Electromyography and magnetic resonance imaging data were collected from normal and atrophied muscles. Electromyography and magnetic resonance imaging abnormalities compatible with denervation atrophy were detected. Although we cannot rule out aging or other unknown causes, we suspect that denervation is caused by nerve damage from trauma to the C1 dorsal ramus as a consequence of entrapment within the rectus capitis posterior major muscle. PMID- 9862544 TI - Sudden death in the dysphagic stroke patient--a case of airway obstruction caused by a food bolus: a brief report. AB - The clinical events leading up to the sudden death of a dysphagic stroke patient with dementia is described. A 63-yr-old man sustained right thalamic and mid brain infarctions. On the inpatient stroke rehabilitation ward, he exhibited significant impulsivity and dementia, the latter felt to be premorbid. The patient frequently coughed, and modified barium swallow testing showed dysphagia, with aspiration occurring only when consuming greater than teaspoon amounts of liquid. He subsequently died at home while eating a meal. Autopsy showed an intact large cheese ball (bocconcini) occluding the airway. Sudden death in the impulsive stroke patient secondary to airway occlusion by a food bolus has not previously been reported, although such patients seem to be at greater risk. New eating-related interventions are warranted for dysphagic patients who exhibit impulsivity. It is proposed that food particle size be limited to 1 cm2 and that such patients be closely monitored while eating. PMID- 9862545 TI - Paravertebral muscle metastases as imaged by magnetic resonance venography: a brief report. AB - Paraspinal muscle metastasis as initially suggested by an electromyographic pattern of isolated posterior primary ramus denervation and subsequently confirmed by magnetic resonance imaging has been reported. However, despite widespread systemic tumor dissemination, metastases to other skeletal muscle occurs infrequently. Uniquely, the paraspinal muscles are drained by the paravertebral plexus of veins. Valveless and at very low pressures, they communicate directly by collaterals with the portal system. Valsalva maneuvers with sudden increases of pressure within the intra-abdominal and intrathoracic cavities can force venous blood from the systemic circulation into the paravertebral plexus of veins. These same venous surges potentially carry tumor emboli to the vertebrae and/or from the vertebral medulla to the adjacent paravertebral muscle by the venous communicators. The inherent increased vascularity of metastatic tumor relative to the surrounding paraspinal muscle as demonstrated by magnetic venous angiography for the first time now permits earlier confirmation and biopsy of the electromyographic-suspected metastatic lesion. In this reported instance of a magnetic resonance imaging-recognized primary lung metastasis confirmed by magnetic resonance venography, there is the future promise of identifying earlier and smaller lesions by this technique. PMID- 9862546 TI - Facilitating patient learning during medical rehabilitation: a research agenda. AB - Although patient learning is widely acknowledged to be an integral part of many medical rehabilitation practices, it has been the subject of little systematic research. A workshop conducted August 18 to 19, 1997, was organized by the National Center for Medical Rehabilitation Research (National Institute of Child Health and Human Development, National Institutes of Health) and several co sponsoring organizations to formulate recommendations concerning learning oriented rehabilitation practices. The recommendations and their supporting rationale are summarized in the topic areas of motor learning and control, cognitive learning, recovery of functioning, generalization and transfer of training, and applications for patients with strokes, traumatic brain injury, amputations, and infants and children. PMID- 9862547 TI - Posttraumatic fibromyalgia at pain facilities versus rheumatologists' offices: a commentary. PMID- 9862548 TI - Comparing visual inspection with statistical analysis of single-subject data. PMID- 9862549 TI - Depressive symptomatology and incident cognitive decline in an elderly community sample. AB - BACKGROUND: It is not known whether depression is a cause or consequence of progressive cognitive decline. We assessed the relationship between depressive symptoms and subsequent cognitive decline in the community-dwelling elderly population. METHODS: Data were from a population-based cohort study that enrolled 2812 noninstitutionalized elderly residents of New Haven, Conn, and followed them with in-home visits in 1982, 1985, 1988, and 1994. Cognitive function was assessed with the Short Portable Mental Status Questionnaire (SPMSQ). Response to the SPMSQ was scored as high, medium, and low, and cognitive decline was defined as a transition to a lower category. Depressive symptoms were measured with the Center for Epidemiological Studies Depression Scale. RESULTS: An elevated level of depressive symptoms was associated with an increased risk of incident cognitive decline among medium SPMSQ performers (3-year odds ratio [OR], 1.72; 95% confidence interval [CI], 1.04-2.82, P=.03; 6-year OR, 2.40; 95% CI, 1.33 4.34; P=.004; 12-year OR, 1.65; 95% CI, 0.62-4.38; P=.31) but not among high performers (3-year OR, 0.93; 95% CI, 0.62-1.39; P=.71; 6-year OR, 1.03; 95% CI, 0.67-1.58; P=.90; 12-year OR, 1.26; 95% CI, 0.59-2.71; P=.55), after adjustment for age, sex, race, education, income, housing type, functional disability, cardiovascular profile, and alcohol use. CONCLUSIONS: Depressive symptoms, particularly dysphoric mood, presage future cognitive losses among elderly persons with moderate cognitive impairments. However, the data do not provide support for the hypothesis that depressive symptoms are associated with the onset or rate of cognitive decline among cognitively intact elderly persons. PMID- 9862550 TI - The depression-dementia conundrum: integrating clinical and epidemiological perspectives. PMID- 9862551 TI - Regional gray matter, white matter, and cerebrospinal fluid distributions in schizophrenic patients, their siblings, and controls. AB - BACKGROUND: Cortical gray matter volume reductions and cerebrospinal fluid (CSF) volume increases are robust correlates of schizophrenia, but their sources have not been established conclusively. METHODS: Structured diagnostic interviews and magnetic resonance imaging scans of the brain were obtained on 75 psychotic probands (63 with schizophrenia and 12 with schizoaffective disorder), ascertained so as to be representative of all such probands in a Helsinki, Finland, birth cohort; 60 of their nonpsychotic full siblings; and 56 demographically similar control subjects without a personal or family history of treated psychiatric morbidity. RESULTS: Patients with schizophrenia and their siblings exhibited significant reductions in cortical gray matter volume and significant increases in sulcal CSF volume compared with controls. The patients, but not their siblings, also exhibited significant reductions in white matter volume and significant increases in ventricular CSF volume. Regional effects were most robust when component volumes were expressed as percentages of overall regional volumes; in this case, for patient and sibling groups, gray matter volume reductions and sulcal CSF volume increases were significantly more pronounced in the frontal and temporal lobes than in the remainder of the brain. None of the group differences varied significantly by sex or hemisphere. CONCLUSIONS: Structural alterations of the cerebral cortex, particularly in the frontal and temporal lobes, are present in patients with schizophrenia and in some of their siblings without schizophrenia; such changes are thus likely to reflect genetic (or shared environmental) effects. Ventricular enlargement is unique to the clinical phenotype and is thus likely to be affected primarily by nonshared causative factors. PMID- 9862552 TI - Word and tone working memory deficits in schizophrenia. AB - BACKGROUND: Verbal memory deficits have been reported in many studies of patients with schizophrenia. We evaluated the specificity of these deficits by comparing patients and control subjects on several verbal and nonverbal auditory memory tests. METHODS: Performance of stable, medicated outpatients with DSM-III-R diagnoses of schizophrenia (N = 38) was compared with that of healthy subjects (N = 39) on a word list immediate recall task, tone delayed discrimination tasks, and word and tone serial position tasks. Before memory testing, patients were divided into 2 groups based on their ability to perform normally on a screening test requiring pitch discrimination and sustained attention. RESULTS: The nonverbal tests were more difficult for control subjects than the verbal tests. Despite this, patients who performed normally on the screening test of perception and attention performed normally on both nonverbal tests but had highly significant deficits on both verbal tests (P<.001 and P = .02). Patients who performed poorly on the screening test had highly significant performance deficits on all the memory tests. CONCLUSIONS: One subgroup of patients with schizophrenia has a selective deficit in verbal memory despite normal motivation, attention, and general perceptual function. Another group has deficits in multiple aspects of cognitive function suggestive of failure in early stages of information processing. PMID- 9862553 TI - Cortical dysfunction in schizophrenia during auditory word and tone working memory demonstrated by functional magnetic resonance imaging. AB - BACKGROUND: Verbal learning and memory deficits are among the most severe cognitive deficits observed in schizophrenia. We have demonstrated that such deficits do not extend to working memory for tones in a substantial number of patients even when verbal working memory is impaired. In this study we used functional magnetic resonance imaging to study the neural basis of this dissociation of auditory verbal and nonverbal working memory in individuals with schizophrenia. METHODS: While undergoing functional magnetic resonance imaging, 12 schizophrenic patients and 12 matched control subjects performed auditory Word Serial Position Task and Tone Serial Position Task. RESULTS: Both tasks produced activation in frontal cortex and temporal and parietal lobes of the cerebrum in both groups. While robust activation was observed in the left inferior frontal gyrus (areas 6, 44, and 45) in the control group during the Word Serial Position Task, activation in the patient group was much reduced in these areas and failed to show the same task-specific activation as in controls. Reduced activation in patients was not confined to the inferior frontal gyrus, but also extended to a medial area during the Tone Serial Position Task and to premotor and anterior temporal lobe areas during both tasks. CONCLUSIONS: These findings support the hypothesis that abnormalities in cortical hemodynamic response in the inferior frontal gyrus underlie the verbal working memory deficit in schizophrenia. The relationship of verbal working memory deficits to other cognitive functions suggests that abnormal functioning in the speech-related areas may reflect a critical substrate of a broad range of cognitive dysfunctions associated with schizophrenia. PMID- 9862554 TI - Functional brain electrical activity mapping in boys with attention deficit/hyperactivity disorder. AB - BACKGROUND: Symptoms of attention-deficit/hyperactivity disorder (ADHD) have been associated with frontal lobe deficits. We used a novel brain electrical imaging method to investigate rapid and continuous changes in brain activity during the continuous performance task (CPT) in normal boys and in boys with ADHD. The amplitude and latency topography of the steady-state visually evoked potential (SSVEP) were examined while subjects performed the "X" version of the CPT (CPT-X; the reference task) and the "A-X" version of the CPT (CPT-AX). METHODS: Seventeen boys meeting DSM-III-R criteria for ADHD and 17 age-matched controls participated in the study. Brain electrical activity was recorded from 64 scalp sites. During the reference task, subjects pressed a microswitch on the unpredictable appearance of the letter X. During the CPT-AX, subjects were required to press the microswitch on the appearance of the letter X only if an A had preceded it. RESULTS: In the interval between the appearances of the A and the X of the correct trials of the CPT-AX, control boys showed transient reductions in SSVEP latency at right prefrontal sites. By contrast, boys with ADHD showed no change or an increase in prefrontal SSVEP latency at right prefrontal sites. CONCLUSION: Our results suggest increased speed of prefrontal neural processing in children without ADHD following a priming stimulus, and a deficit in such processes in children with ADHD. PMID- 9862555 TI - Family history of alcoholism and hypothalamic opioidergic activity. AB - BACKGROUND: This study was designed to assess whether nonalcoholic offspring from families with a high density of alcohol-dependent individuals have altered endogenous central nervous system opioid activity. Naloxone hydrochloride stimulates plasma cortisol by blocking opioidergic input on the corticotropin releasing factor neuron, thereby providing a noninvasive method for measuring hypothalamic opioid tone. METHODS: Forty-eight nonalcoholic subjects aged 18 to 25 years were enrolled in a protocol to measure endogenous opioid activity by inducing opioid receptor blockade with the receptor antagonist, naloxone. Twenty six subjects were offspring from families with a high density of alcohol dependence and were designated as family history-positive subjects. Twenty-two subjects were biological offspring of nonalcohol-dependent parents and designated as family history-negative subjects. Subjects received naloxone hydrochloride (0, 125, and 375 microg/kg) in double-blind, randomized order. Serum cortisol levels were monitored. RESULTS: Family history-negative subjects had a graded cortisol response to each dose of naloxone. In contrast, family history-positive subjects achieved a maximal cortisol response to the 125-microg/kg dose of naloxone hydrochloride with no further increase in cortisol levels observed following the 375-microg/kg dose. Family history-negative subjects had a diminished cortisol response to the 125-microg/kg dose compared with the family history-positive subjects. Plasma naloxone concentrations did not differ between family history groups. CONCLUSIONS: Individuals from families with a high density of alcohol dependence are more sensitive to naloxone compared with offspring of nonalcohol dependent parents. This implies that individuals with a family history of alcohol dependence have diminished endogenous hypothalamic opioid activity. PMID- 9862556 TI - Treating major depression in primary care practice: an update of the Agency for Health Care Policy and Research Practice Guidelines. AB - The Depression Guideline Panel of the Agency for Health Care Policy and Research in 1993 published recommendations for treating major depression in primary care practice that were often based on studies of tertiary care psychiatric patients. We reviewed reports of randomized controlled trials in primary care settings published between 1992 and 1998. This evidence indicates that both antidepressant pharmacotherapy and time-limited depression-targeted psychotherapies are efficacious when transferred from psychiatric to primary care settings. In most cases, the choice between these treatments should depend on patient preference. Studies to date suggest that improving treatment of depression in primary care requires properly organized treatment programs, regular patient follow-up, monitoring of treatment adherence, and a prominent role for the mental health specialist as educator, consultant, and clinician for the more severely ill. Future research should focus on how guidelines are best implemented in routine practice, since conventional dissemination strategies have little impact. PMID- 9862557 TI - The effects of adherence to antidepressant treatment guidelines on relapse and recurrence of depression. AB - BACKGROUND: Depression is associated with high rates of relapse and recurrence during a patient's lifetime. Current guidelines regarding treatment recommend 4 to 9 months of continuation antidepressant therapy following remission of acute symptoms to allow more complete resolution of the episode. In this article, we test whether adherence to these recommendations reduces the likelihood of relapse or recurrence in a Medicaid population. METHODS: We used a Medicaid database covering 1989 through 1994. The sample consists of the 4052 adult patients who filled an antidepressant prescription at the time of an initial diagnosis of depression. These patients were followed up for up to 2 years. Timing and counts of antidepressant prescription claims are used to construct a proxy measure for adherence to guidelines. Relapse or recurrence is defined by evidence of a new episode requiring antidepressant treatment, hospital admission for depression, electroconvulsive therapy, emergency department visit for mental health, or attempted suicide. We used survival analysis to predict relapse or recurrence for each patient and to examine the effect of following treatment guidelines on relapse and recurrence. RESULTS: Approximately one fourth of the patients had a relapse or recurrence during their follow-up period. Factors that affect relapse and recurrence include comorbidities, race, and guideline adherence. Those who continued therapy with their initial antidepressant were least likely to experience relapse or recurrence; those who discontinued their antidepressant early were most likely to experience relapse or recurrence. CONCLUSION: Adherence to depression treatment guidelines with an antidepressant that is likely to have continuous use by patients reduces the probability of relapse or recurrence. PMID- 9862559 TI - "Clinical significance" and DSM-IV. PMID- 9862558 TI - Cognitive behavioral group therapy vs phenelzine therapy for social phobia: 12 week outcome. AB - BACKGROUND: This article presents results of the acute treatment phase of a 2 site study comparing cognitive behavioral group therapy (CBGT) and treatment with the monoamine oxidase inhibitor phenelzine sulfate for social phobia. METHODS: One hundred thirty-three patients from 2 sites received 12 weeks of CBGT, phenelzine therapy, pill placebo administration, or educational-supportive group therapy (an attention-placebo treatment of equal credibility to CBGT). The "allegiance effect," ie, the tendency for treatments to seem most efficacious in settings of similar theoretical orientation and less efficacious in theoretically divergent settings, was also examined by comparing responses to the treatment conditions at both sites: 1 known for pharmacological treatment of anxiety disorders and the other for cognitive behavioral treatment. RESULTS: After 12 weeks, phenelzine therapy and CBGT led to superior response rates and greater change on dimensional measures than did either control condition. However, response to phenelzine therapy was more evident after 6 weeks, and phenelzine therapy was also superior to CBGT after 12 weeks on some measures. There were few differences between sites, suggesting that these treatments can be efficacious at facilities with differing theoretical allegiances. CONCLUSIONS: After 12 weeks, both phenelzine therapy and CBGT were associated with marked positive response. Although phenelzine therapy was superior to CBGT on some measures, both were more efficacious than the control conditions. More extended cognitive behavioral treatment and the combination of modalities may enhance treatment effect. PMID- 9862560 TI - Limitations of diagnostic paradigm: it doesn't explain "need". PMID- 9862562 TI - Wild-type p53-mediated down-modulation of interleukin 15 and interleukin 15 receptors in human rhabdomyosarcoma cells. AB - We recently reported that rhabdomyosarcoma cell lines express and secrete interleukin 15 (IL-15), a tightly regulated cytokine with IL-2-like activity. To test whether the p53-impaired function that is frequently found in this tumour type could play a role in the IL-15 production, wild-type p53 gene was transduced in the human rhabdomyosarcoma cell line RD (which harbours a mutated p53 gene), and its effect on proliferation and expression of IL-15 was studied. Arrest of proliferation was induced by wild-type p53; increased proportions of G1-arrested cells and of apoptotic cells were observed. A marked down-modulation of IL-15 expression, at both the mRNA and protein level, was found in p53-transduced cells. Because a direct effect of IL-15 on normal muscle cells has been reported, the presence of IL-15 membrane receptors was studied by cytofluorometric analysis. Rhabdomyosarcoma cells showed IL-15 membrane receptors, which are down modulated by wild-type p53 transfected gene. In conclusion, wild-type p53 transduction in human rhabdomyosarcoma cells induces the down-modulation of both IL-15 production and IL-15 receptor expression. PMID- 9862561 TI - Midtown Manhattan prevalence rates and the implied need for treatment: meeting the challenge of public mental health. PMID- 9862563 TI - Hypoxia induces p53 accumulation in the S-phase and accumulation of hypophosphorylated retinoblastoma protein in all cell cycle phases of human melanoma cells. AB - Hypoxia has been shown to induce accumulation of p53 and of hypophosphorylated retinoblastoma protein (pRb) in tumour cells. In this study, the cell cycle dependence of p53 accumulation and pRb hypophosphorylation in four human melanoma cell lines that are wild type for p53 was investigated using two-parameter flow cytometry measurements of p53 or pRb protein content and DNA content. The hypoxia induced increase in p53 protein was higher in S-phase than in G1 and G2 phases in all cell lines. The accumulation of p53 in S-phase during hypoxia was not related to hypoxia-induced apoptosis or substantial cell cycle specific cell inactivation during the first 24 h of reoxygenation. pRb was hypophosphorylated in all cell cycle phases by hypoxia treatment. The results did not support a direct link between p53 and pRb during hypoxia because p53 was induced in a cell cycle specific manner, whereas no cell cycle-dependent differences in pRb hypophosphorylation were detected. Only a fraction of the cell populations (0.60+/-0.10) showed hypophosphorylated pRb. Thus, pRb is probably not the only mediator of the hypoxia-induced cell cycle block seen in all cells and all cell cycle phases. Moreover, the cell cycle-dependent induction of p53 by hypoxia suggests that the primary function of p53 accumulation during hypoxia is other than to arrest the cells. PMID- 9862564 TI - Radiation-induced micronuclei in human fibroblasts in relation to clonogenic radiosensitivity. AB - As part of our programme for developing predictive tests for normal tissue response to radiotherapy, we have investigated the efficacy of the cytokinesis block micronucleus (MN) assay as a means of detecting interindividual differences in cellular radiosensitivity. A study was made of nine fibroblast strains established from vaginal biopsies of pretreatment cervical cancer patients and an ataxia telangiectasia (A-T) cell strain. Cells were irradiated in plateau phase, replated and treated with cytochalasin B 24 h later. MN formation was examined 72 h after irradiation as the number of MN in 100 binucleate cells. The method yielded low spontaneous MN yields (<7 per 100 cells), and mean induced MN frequencies after 3.5 Gy varied between cell strains from 18 to 144 per 100 cells. However, in repeat experiments, considerable intrastrain variability was observed (CV = 32%), with up to twofold differences in MN yields, although this was less than interstrain variability (CV = 62%). An analysis was made of the relationship between MN results and previously obtained clonogenic survival data. There was a significant correlation between MN yields and clonogenic survival. However, when the A-T strain was excluded from the analysis, the correlation lost significance, mainly because of one slow-growing strain which was the most sensitive to cell killing but had almost the lowest MN frequency. With current methodology, the MN assay on human fibroblasts does not appear to have a role in predictive testing of normal tissue radiosensitivity. PMID- 9862565 TI - A metalloprotease activity from C6 glioma cells inactivates the myelin-associated neurite growth inhibitors and can be neutralized by antibodies. AB - Glioblastoma cells infiltrate brain tissue and migrate preferentially along white matter fibre tracts, an environment that is highly inhibitory to the migration of astrocytes and the growth of neurites because of the presence of specific inhibitory proteins. In vitro, spreading and migration of rat C6 glioma cells on a CNS (central nervous system) myelin substrate is correlated with and dependent on the presence of a metalloprotease. This membrane-bound metalloendoprotease exhibits a blocker profile different from known proteases. Pretreatment of CNS myelin or of a highly purified CNS myelin component, the inhibitory protein bNI 220, with C6 metalloproteolytic activity converts these non-permissive substrates into permissive environments for astrocytes and fibroblasts, indicating that this C6 cell-derived metalloprotease may inactivate myelin-associated inhibitory proteins. Antibodies were raised in chicken against fractions enriched in metalloproteolytic activity; these antibodies were able to inhibit specifically spreading and migration of C6 glioma cells on a CNS myelin substrate, as well as the invasion of C6 cells into adult rat optic nerve explants in vitro. These results suggest a crucial involvement of a membrane-bound metalloprotease in the mechanisms of C6 glioma migration and infiltration of brain tissue by proteolytic inactivation of the neurite growth inhibitory proteins present in CNS myelin. PMID- 9862566 TI - Relationship between the expression of vascular endothelial growth factor and the density of dendritic cells in gastric adenocarcinoma tissue. AB - It has been reported that decreased numbers of dendritic cells (DCs) are correlated with poor prognosis in some types of malignancy, such as gastric cancer. However, factors that determine the density of DCs have not been characterized. It was recently reported that vascular endothelial growth factor (VEGF) inhibits the functional maturation of DCs from CD34+ precursors. In this study, we analysed the relationship between the expression of VEGF and the density of DCs in gastric carcinoma tissues by immunohistochemical staining. The extent of infiltration by DCs was graded from marked to slight on the basis of the mean densities of DCs. The prognosis of patients with marked infiltration was significantly better than that of patients with slight infiltration among patients who had undergone curative resection. Multivariate analysis showed that infiltration by DCs was an independent prognostic indicator. Furthermore, there was an inverse correlation between the density of DCs and the expression of VEGF Our results suggest that expression of VEGF might be associated with tumour progression and poor prognosis not only because VEGF stimulates angiogenesis, but also because it allows tumours to escape from attack by the immune system in patients with gastric carcinoma. PMID- 9862568 TI - Characterization of human soft-tissue sarcoma xenografts for use in secondary drug screening. AB - We have established ten transplantable human soft-tissue sarcoma (STS) xenografts grown as subcutaneous tumours in the nude mouse. Nine xenografts originated from patients that needed chemotherapy in the course of their disease. The xenografts were tested for their sensitivity to maximum tolerated doses of five anti-cancer agents. Growth of treated tumours was expressed as a percentage of control tumour growth and a growth inhibition > 75% was measured for doxorubicin in 20% of the STS xenografts, for cyclophosphamide in 30%, for ifosfamide in 20%, for vincristine in 20%, whereas etoposide was not effective in the STS xenografts. In three out of ten STS xenografts MDR1 mRNA was detectable, but this was not related to the resistance against doxorubicin, vincristine or etoposide. Topoisomerase IIalpha mRNA expression levels did not reflect sensitivity to doxorubicin or etoposide. In all STS tissues, however, these levels were lower than topoisomerase IIalpha mRNA in a drug-sensitive human ovarian cancer xenograft. Glutathione concentrations and the activities of glutathione S transferase, glutathione peroxidase and glutathione reductase were not related to resistance against the alkylating agents or doxorubicin. Of interest, in all STS tissues, glutathione S-transferase pi was the predominant isoenzyme present. In conclusion, chemosensitivity of the STS xenografts reflects clinical response rates in phase II trials on the same compounds in adult STS patients. Relatively low levels of topoisomerase IIalpha mRNA may partly account for intrinsic resistance against, for example, doxorubicin. Additional factors must contribute to moderate responsiveness to alkylating agents. PMID- 9862567 TI - In vitro inhibition of human malignant brain tumour cell line proliferation by anti-urokinase-type plasminogen activator monoclonal antibodies. AB - A brain tumour-associated marker, urokinase (UK), was investigated using rabbit anti-UK polyclonal and murine anti-UK monoclonal antibodies, which were prepared by immunization with low molecular weight UK (LMW-UK) and high molecular weight urokinase (HMW-UK) synthetic peptide respectively. The polyclonal antibody cross reacted with both LMW-UK and HMW-UK, whereas the murine MAbs were specific for HMW-UK. These immunological probes were used to study urokinase in glioma extracts, tissues, sera and cell lines that had been prepared from primary cultures of freshly dissected gliomas. Radioimmunoassays showed that glioma extracts had much higher level (5- to 44-fold) of UK than normal human brain extracts. This result was confirmed by immunoblotting of electrophoresis gels of glioma and human brain extracts. Immunohistochemical study using anti-UK MAb demonstrated much higher levels of UK in glioma tissue than normal brain tissue. Immunohistochemical study using anti-UK MAbs localized UK on the cell surface of glioma cells. Anti-UK MAbs inhibited the proliferation of AA cell lines and GB cell lines (50% to > 90%) and exerted minor effects (< or = 20%) on normal human liver, intestine and lymphocyte cell lines. Taken together, these results suggest that anti-UK MAbs may have therapeutic potential for human gliomas and cancer metastasis. PMID- 9862569 TI - Apoptosis after gamma irradiation. Is it an important cell death modality? AB - Apoptosis and necrosis are two different forms of cell death that can be induced by cytotoxic stress, such as ionizing radiation. We have studied the importance of apoptotic death induced after treatment with 6 Gy of gamma-irradiation in a panel of eight human tumour cell lines of different radiosensitivities. Three different techniques based on the detection of DNA fragmentation have been used, a qualitative one--DNA ladder formation --and two quantitative approaches--in situ tailing and comet assay. No statistically significant relationship between the two quantitative assays was found (r= 0.327, P = 0.159) so these methods seem to show different aspects of the process of cell death. The presence of the DNA ladder related well to the end-labelling method in that the least amount of end labelling was seen in samples in which necrotic degradation rather than apoptotic ladders were seen. However, as the results obtained by the comet assay are not in agreement with the DNA ladder experiments, we suggest that the distinction between the degraded DNA produced by apoptosis and necrosis may be difficult by this technique. Finally, although apoptosis has been proposed to be dependent on p53 functionality, and this may explain differences in cellular radiosensitivity, no statistically significant relationship was found between these parameters and apoptosis in the eight cell lines studied. PMID- 9862570 TI - The efficacy of the anthracycline prodrug daunorubicin-GA3 in human ovarian cancer xenografts. AB - The prodrug N-[4-(daunorubicin-N-carbonyl-oxymethyl)phenyl] O-beta-glucuronyl carbamate (DNR-GA3) was synthesized for specific activation by human beta glucuronidase, released in necrotic areas of tumour lesions. In vitro, DNR-GA3 was 18 times less toxic than daunorubicin (DNR) and the prodrug was completely activated to the parent drug by human beta-glucuronidase. The maximum tolerated dose of DNR-GA3 in nude mice bearing s.c. human ovarian cancer xenografts was 6 10 times higher than that of DNR. The prodrug was cleared more rapidly from the circulation (elimination t1/2 = 20 min) than the parent drug (elimination t1/2 = 720 min). The anti-tumour effects of DNR-GA3 and DNR were investigated in four different human ovarian cancer xenografts OVCAR-3, FMa, A2780 and MRI-H-207 at a mean tumour size between 100 and 200 mm3. In three out of four of these tumour lines, the prodrug given i.v. at the maximum tolerated dose ranging from 150 to 250 mg kg(-1) resulted in a maximum tumour growth inhibition from 82% to 95%. The standard treatment with DNR at a dose of 8 mg kg(-1) given i.v. weekly x 2 resulted only in a maximum tumour growth inhibition from 40% to 47%. Tumour line FMa did not respond to DNR, nor to DNR-GA3. Treatment with DNR-GA3 was also given to mice with larger tumours that would contain more necrosis (mean size 300-950 mm3). The specific growth delay by DNR-GA3 was extended from 2.1 to 4.4 in OVCAR 3 xenografts and from 4.4 to 6.0 in MRI-H-207 xenografts. Our data indicate that DNR-GA3 is more effective than DNR and may be especially of use for treatment of tumours with areas of necrosis. PMID- 9862571 TI - Low-density lipoprotein receptor-mediated delivery of a lipophilic daunorubicin derivative to B16 tumours in mice using apolipoprotein E-enriched liposomes. AB - Many tumours express relatively high levels of low-density lipoprotein (LDL) receptors on their membranes. The LDL receptor is, therefore, an attractive target for the selective delivery of antineoplastic drugs to tumour cells. We reported previously on the synthesis of small apolipoprotein E (apoE)-containing liposomes that behave in vivo in a very similar way to native LDL. In this study, we examined the interaction of this liposomal carrier with cultured B16 melanoma cells. Binding of apoE liposomes to the cells is saturable, with a maximum binding of approximately 90000 particles per cell. Cross-competition studies indicated that apoE liposomes are bound by the LDL receptor. Association of apoE liposomes to B16 cells is strictly Ca2+ dependent, which forms additional evidence for a role of the LDL receptor. The affinity of apoE liposomes for the LDL receptor on B16 cells is 15-fold higher than that of LDL (0.77 vs 11.5 nM respectively). ApoE is essential for the LDL receptor recognition because liposomes lacking apoE were, in competition studies, 20- to 50-fold less effective than apoE-containing liposomes. We examined in B16 tumour-bearing mice the tumour-localizing properties of apoE liposomes and the disposition of an incorporated lipophilic derivative of daunorubicin (LAD). Tissue distribution studies showed that LAD-loaded apoE liposomes were taken up and processed by the major LDL receptor-expressing organs (i.e. adrenals, liver and spleen). Of all other tissues, the tumour showed the highest uptake. The distribution patterns of LAD-loaded apoE liposomes and native LDL in the tumour-bearing mice were very similar, which supports the role of the LDL receptor in the disposition of the prodrug-loaded particles. The disposition of LAD followed the pattern of the liposomal carrier. We conclude that apoE liposomes enable LDL receptor-mediated specific delivery of antineoplastic (pro)drugs to tumours, and, therefore, constitute an attractive novel option for anti-tumour chemotherapy. PMID- 9862572 TI - Smad4 (DPC4)--a potent tumour suppressor? AB - The recently described family of Smad molecules are essential mediators of transforming growth factor beta (TGF-beta) signalling. To date, seven members of this family have been identified, each of which plays a specific and separate role in mediating TGF-beta superfamily gene transcription. At least two different Smads, Smad2 and Smad4 (DPC4), have been implicated in human cancer and appear to have tumour-suppressor functions. Loss of function of Smad4 is most strongly associated with human pancreatic and colorectal malignancy. Furthermore, work from several different groups has suggested associations between Smad4 loss and malignancy in a number of other tissues. Here, we present a review of the current state of the literature implicating the central Smad mediator, Smad4, in the development of cancer. PMID- 9862575 TI - Radiobiological prediction of normal tissue toxicities and tumour response in the radiotherapy of advanced non-small-cell lung cancer. AB - A number of randomized studies have been carried out in the UK and USA to determine the optimal radiotherapy dose schedule for advanced non-small-cell lung cancer (NSCLC). We have examined eight radiotherapy regimens from data taken from four randomized phase III studies carried out in the UK (1264 patients): 10 Gy single fraction; 17 Gy in two fractions over 8 days; 30 Gy in ten fractions over 14 days; 22.5 Gy in five fractions in 5 days; 27 Gy in six fractions over 11 days; 30 Gy in six fractions over 11 days; 36 Gy in 12 fractions over 16 days; and 39 Gy in 13 fractions over 17 days. We compared the clinical results in palliation, toxicity and survival with four regimens taken from one randomized study from the USA (365 patients): 40 Gy in 20 fractions over 4 weeks; 40 Gy 'split course' in ten fractions in 4 weeks; 50 Gy in 25 fractions over 5 weeks; and 60 Gy in 30 fractions over 6 weeks. Using the linear-quadratic (LQ) radiobiological model, we have calculated the radiobiological equivalent dose (BED) for acute-reacting tissues (BED10), late-reacting tissues (BED1.7) and tumour (BED25), and related the predicted response to the observed response in each tissue. There was a good correlation between the predicted response and the reported response in the case of late-reacting tissue toxicity and tumour response. The model confirmed that, in good performance status patients, a higher value for BED25 correlated with a higher degree of local control and survival and that radiotherapy regimens with a higher value for BED1.7 were associated with five cases of cord myelopathy, if the spinal cord was not shielded. In poor performance status patients the model suggested that the optimal regimen was a single fraction of 10 Gy because this resulted in an equivalent degree of symptom control as other regimens, needed only one hospital visit and was less likely to result in cord damage, thus, allowing for the possibility of retreatment at a later date. PMID- 9862573 TI - A feasibility study of roquinimex (Linomide) and alpha interferon in patients with advanced malignant melanoma or renal carcinoma. AB - Thirty-one patients with advanced renal carcinoma or malignant melanoma were treated in the first feasibility study of alpha-interferon (Roferon) and the new oral immunomodulating agent, Linomide. Linomide 5 mg or 10 mg p.o. daily was given for 2 weeks; alpha-interferon was then added at 3 MU s.c. three times weekly, escalating in each patient by 3 MU per week, if tolerable, up to 12 MJ. The combination was poorly tolerated with nausea, vomiting, somnolence and myalgia commonly reported. Adverse events accounted for treatment withdrawal in ten patients and contributed to withdrawal in four other patients. Treatment with Linomide alone in the first 2 weeks led to a significant increase in white blood cells, neutrophils and platelets. When alpha-interferon was added, the platelet count decreased significantly over the following 6 weeks. Nineteen patients had white cell phenotype and function measured. After 2 weeks of 5 mg Linomide, a transient but significant decrease in the absolute number of activated T-helper cells (CD4+DR+) was observed. No changes in natural killer (NK) cell number or activity were observed. Twenty-two patients were evaluable for response. One with metastatic renal cell carcinoma had a complete response and six had stable disease. This study does not support the use of the combination because significant toxicity was seen without the anticipated immunological benefits. PMID- 9862574 TI - Adjuvant chemotherapy for primary cardiac sarcomas: the IGR experience. AB - The effect of additional treatments after surgery in patients with primary cardiac sarcoma (PCS) remains unknown. The present study aims to evaluate the benefit of chemotherapy in patients with non-metastatic cardiac sarcomas after optimal resection. Between October 1979 and December 1995, 15 patients with a median age of 45 (range 16-66) and a resected primary cardiac sarcoma [angiosarcoma (six), malignant fibrous histiocytoma (three), leiomyosarcoma (two), rhabdomyosarcoma (two), liposarcoma (one) and synoviosarcoma (one)] received a doxorubicin-containing regimen within 6 weeks of surgery. Adjuvant chemotherapy combinations included cyclophosphamide, vincristine and dacarbazine in four patients; ifosfamide in nine; methotrexate and vincristine in one; and doxorubicin alone in one patient. At present, 13 patients have relapsed (five during therapy), with a median time to progression of 10 months. Twelve patients developed local relapse, in four cases without metastatic disease. Two patients remain in complete remission 27 and 25 months after surgery. The median time to progression was shorter in patients presenting a cardiac angiosarcoma than other histological types (3 vs 14 months, P < 0.01). Twelve patients have died, with a median overall survival of 12 months. The 2-year survival rate is 26%. Survival was significantly longer for patients with completely resected tumours (22 vs 7 months; P = 0.02) and those who did not have angiosarcoma (18 vs 7 months; P = 0.04). In conclusion, post-operative conventional doxorubicin-based chemotherapy failed to modify the natural history of patients with resected cardiac sarcomas. Locoregional failure remains the main problem even after histologically complete resection. New approaches must be tested in patients with primary cardiac sarcoma. PMID- 9862576 TI - High-dose epirubicin is not an alternative to standard-dose doxorubicin in the treatment of advanced soft tissue sarcomas. A study of the EORTC soft tissue and bone sarcoma group. AB - The activity and toxicity of single-agent standard-dose doxorubicin were compared with that of two schedules of high-dose epirubicin. A total of 334 chemonaive patients with histologically confirmed advanced soft-tissue sarcomas received (A) doxorubicin 75 mg m(-2) on day 1 (112 patients), (B) epirubicin 150 mg m(-2) on day 1 (111 patients) or (C) epirubicin 50 mg m(-2) day(-1) on days 1, 2 and 3 (111 patients); all given as bolus injection at 3-week intervals. A median of four treatment cycles was given. Median age was 52 years (19-70 years) and performance score 1 (0-2). Of 314 evaluable patients, 45 (14%) had an objective tumour response (eight complete response, 35 partial response). There were no differences among the three groups. Median time to progression for groups A, B and C was 16, 14 and 12 weeks, and median survival 45, 47 and 45 weeks respectively. Neither progression-free (P = 0.93) nor overall survival (P = 0.89) differed among the three groups. After the first cycle of therapy, two patients died of infection and one owing to cardiovascular disease, all on epirubicin. Both dose schedules of epirubicin were more myelotoxic than doxorubicin. Cardiotoxicity (> or = grade 3) occurred in 1%, 0% and 2% respectively. Regardless of the schedule, high-dose epirubicin is not a preferred alternative to standard-dose doxorubicin in the treatment of patients with advanced soft tissue sarcomas. PMID- 9862577 TI - Expression of a truncated form of hHb1 hair keratin in human breast carcinomas. AB - Human hHb1 belongs to the type II hard keratin family and is physiologically expressed in hair shafts. In the present study, using specific 3' and 5' probes for hHb1, we established that breast carcinomas ectopically express a hHb1 5' truncated mRNA, and that this transcript is restricted to malignant epithelial cells. Furthermore, an in vitro study indicated that it could be translated. We concluded that, in breast carcinomas, expression of truncated hHb1 is related to epithelial cell transformation. Because the hHb1 gene maps to 12q11-q13, a chromosome region known to present several breakpoints in solid tumours, we propose that the hHb1 gene might represent a target for such alterations. PMID- 9862578 TI - Immunoradiometric and immunohistochemical analysis of Cathepsin D in ovarian cancer: lack of association with clinical outcome. AB - The aim of this study was to analyse the clinical significance of Cathepsin D (Cath D) content as determined by an immunoradiometric assay in a series of primary untreated ovarian cancers from 162 patients. In addition, immunohistochemical analysis of Cath D was also performed on a subset of 86 tumours. Cath D levels were distributed in an asymmetrical way and were skewed towards the lower values (median value 20.8 pmol mg(-1) protein, range 2.0-99.0 pmol mg(-1) protein). No correlation was found between Cath D levels and clinicopathological parameters. However, the percentage of Cath D positivity was significantly higher in oestrogen receptor-positive (57%) compared with oestrogen receptor-negative (36%) cases (P= 0.01). The percentage of Cath D-positive staining was not significantly different for both epithelial (27%) and stromal components (40%). Immunoradiometrically detected Cath D levels were not different according to Cath D stromal immunostaining (P= 0.18), while higher Cath D levels were measured in Cath D-positive than in Cath D-negative tumour epithelial cells (P = 0.027). Survival analysis was conducted on 161 primary untreated ovarian cancer patients. The 5-year overall survival rate was 57% and 55% in Cath D positive and Cath D-negative patients respectively (P = 0.69). As far as time to progression was concerned, there was no significant difference in the survival rate of patients with either high or low Cath D content (P = 0.56). Similar results have been obtained in the subset of patients in which Cath D was analysed by immunohistochemistry. In conclusion, Cath D measurement in tumour extracts appears to have a limited usefulness in improving the prognostic characterization of ovarian cancer patients. PMID- 9862579 TI - Chromosome 5 allelic losses are early events in tumours of the papilla of Vater and occur at sites similar to those of gastric cancer. AB - During our studies of DNA fingerprinting of tumours of the pancreas and papilla (ampulla) of Vater, using arbitrarily primed polymerase chain reaction (AP-PCR), we noticed two bands showing a decreased intensity in six of ten ampullary tumours with respect to matched normal tissues. Those bands were both assigned to chromosome 5. Such a finding was somewhat in contrast with the reportedly low frequency of APC gene mutations in ampullary cancers, located at chromosome 5q21, and suggested that loci different from that of APC might be the target of chromosome 5 allelic losses (LOH) in these tumours. Therefore, we analysed chromosome 5 LOH in a panel of 27 ampullary tumours, including eight adenomas, four early- and 15 advanced-stage cancers, using 16 PCR-amplified CA microsatellite polymorphic markers spanning the entire chromosome. Nineteen cases (70%) showed LOH, and the interstitial deletions found in these tumours described two smallest common deleted regions, in which putative suppressor genes might reside. They were at 5q13.3-q14 and at 5q23-q31 respectively, which correspond to those found in gastric tumours. In addition, the presence of 5q LOH in six of eight adenomas and in three of four early-stage cancers suggests that such phenomena occur at early stages of neoplastic progression of the ampullary epithelium. PMID- 9862581 TI - You can run, but you can't hide: the academic physician and molecular biology. PMID- 9862580 TI - Aberrant cytoplasmic expression of the p16 protein in breast cancer is associated with accelerated tumour proliferation. AB - The p16 protein plays an important role in the transition of cells into the G1 phase of the cell cycle. We have studied the prevalence of p16 protein expression in breast carcinomas in a prospective series of 368 invasive and 52 non-invasive malignancies, as well as in 88 locally recurring tumours and three tumour cell lines. p16 protein expression was evaluated immunohistochemically on paraffin sections using monoclonal and polyclonal anti-p16 antibodies, and by immunoblotting of tumour cell suspensions. Tumour cell lines were also subjected to polymerase chain reaction-single strand polymorphism (PCR-SSCP) analysis and direct DNA sequencing. The results were compared with established prognostic parameters, DNA flow cytometry and p53 protein expression. In 33 (9%) invasive and two (4%) intraductal carcinomas, a cytoplasmic accumulation of the p16 protein was seen. These cases were characterized by poor histological grade of differentiation, loss of of oestrogen receptors and progesterone receptors and frequent overexpression of the p53 protein. In addition, breast carcinomas with aberrant p16 expression demonstrated a high proliferative activity, with median S phase fractions 74% higher than in the control group and the median Ki67 fractions elevated to 75%. A genetic alteration of the p16 gene was not detectable in three analysed cell lines with cytoplasmic p16 expression applying PCR-SSCP and direct DNA sequencing. These results indicate that cytoplasmic accumulation of the p16 protein identifies a subset of highly malignant breast carcinomas with accelerated tumour proliferation and other unfavourable parameters in breast cancer. The described protein accumulation is apparently not caused by an alteration of the p16 gene. PMID- 9862582 TI - Apoptosis: on the verge of clinical relevance. PMID- 9862583 TI - Apoptosis: only the good die young? PMID- 9862584 TI - Derivation of a clinical prediction model for the emergency department diagnosis of ectopic pregnancy. AB - OBJECTIVE: To derive a clinical prediction model for estimating the pretest probability of ectopic pregnancy in ED patients with first-trimester abdominal pain or vaginal bleeding. METHODS: All hemodynamically stable first-trimester patients presenting to the ED of a tertiary care military teaching hospital over a 14-month period with a chief complaint of abdominal pain and/or vaginal bleeding had clinical data coded prior to determining outcome. They were then followed longitudinally until a criterion standard pregnancy outcome was established. RESULTS: Of the 486 patients enrolled, 280 (58%) had viable intrauterine pregnancies, 167 (34%) had nonviable intrauterine pregnancies, and 39 (8%) had ectopic pregnancies. Using a recursive partitioning model, a high risk group was derived (that was separated from intermediate and low-risk groups), consisting of patients with abdominal peritoneal signs or definite cervical motion tenderness, with a sensitivity of 31% (95% CI: 17-48%), a specificity of 93% (95% CI: 90-95%), a positive likelihood ratio of 4.3, and a negative likelihood ratio of 0.74. A low-risk group, consisting of patients with either fetal heart tones or tissue at the cervical os, or the absence of pain other than midline menstrual-like cramping and lacking any pelvic tenderness, was differentiated from an intermediate-risk group, with a sensitivity of 96% (95% CI: 81-100%), a specificity of 22% (95% CI: 18-26%), a positive likelihood ratio of 1.2, and a negative likelihood ratio of 0.17. CONCLUSION: A clinical prediction model for estimating the probability of ectopic pregnancy in ED patients has been derived. It may prove to have practical clinical application for estimating pretest probability of ectopic pregnancy as well as assisting in medical decision making when laboratory and ultrasonographic findings are nondiagnostic. Clinical application should await prospective validation in an independent sample. PMID- 9862585 TI - Effect of magnesium hydroxide on iron absorption following simulated mild iron overdose in human subjects. AB - OBJECTIVE: To determine the effect of oral magnesium hydroxide [Mg(OH)2] on iron absorption after simulated iron overdose in human subjects. METHODS: A randomized, controlled crossover study was conducted in healthy adult male human volunteers taking no medications. Subjects received an average of 5.0 mg/kg elemental iron orally followed 1 hour later by either oral administration of 4.5 g of Mg(OH)2 per g ingested elemental iron or no treatment. Serial serum specimens were obtained over the 12 hours following iron ingestion and stored at 60 degrees C until standard serum iron assay was performed. After a 2-week washout period, the subjects were enrolled in the alternative trial arm. Individual baseline diurnal variation in serum iron levels was determined over a 12-hour period on the day prior to each trial. Area under time-concentration curves (AUCs) were calculated, and the AUC due to experimental iron ingestion (deltaAUC) was determined by subtracting the baseline diurnal AUC from the experimental AUC for each subject. RESULTS: Thirteen healthy adult male subjects were enrolled. Mean +/- SEM for deltaAUC due to experimental iron ingestion followed by treatment with Mg(OH)2, 78 +/- 23 micromol(hr)/L, was significantly less than that followed by no treatment, 144 +/- 33 micromol(hr)/L (p = 0.03 by signed rank test). CONCLUSIONS: Magnesium hydroxide, administered 1 hour post iron ingestion at an oral dose of 4.5 g per g elemental iron ingested, significantly reduced iron absorption during a 12-hour period following simulated mild iron overdose in healthy adult human volunteers. PMID- 9862586 TI - Effect of skin pigmentation on pulse oximetry accuracy in the emergency department. AB - OBJECTIVE: To determine whether pulse oximeter (PO) accuracy and signal quality are affected by level of skin pigmentation. METHODS: Observational study in a community hospital ED. Consecutive adult patients undergoing arterial blood gas determination were enrolled into the study. Skin pigmentation was determined by comparison with standardized color swatches under controlled lighting; assigned values were used to stratify patients into 3 groups (light, intermediate, and dark) using predetermined criteria. Simultaneous with arterial blood sampling, staff recorded PO reading of O2 saturation using the Nellcor D-25 oximeter. PO values were compared with criterion standard values measured using a 4-wavelength spectrophotometer or co-oximeter. PO signal quality also was recorded. Bias (the mean difference between PO and co-oximeter-measured values of hemoglobin saturation) and precision (the standard deviation of the bias) were calculated. Groups were compared using one-way ANOVA, Bartlett's test for variances, and chi2 test. RESULTS: O2 saturation data were obtained for 284 patients. Bias values did not differ between the 3 skin pigment groups (p = 0.79). Precision was of borderline significance (p = 0.05), but there was no dose-response relation between skin pigmentation and precision. Study personnel reported suboptimal PO function most often among patients in the dark group (p = 0.003), but this finding was of no clinical significance. PO signal failure was rare (<1% of all patients). CONCLUSIONS: Although several prior studies suggest the contrary, this study found that skin pigmentation does not affect the bias or precision of pulse oximetry. Furthermore, skin pigmentation has no clinically significant effect on PO signal quality. PMID- 9862587 TI - Intravenous ketamine for pediatric sedation in the emergency department: safety profile with 156 cases. AB - OBJECTIVES: To determine the safety of i.v. ketamine when administered by emergency physicians (EPs) for pediatric procedures, and to contrast the sedation characteristics of the i.v. and i.m. routes. METHODS: The study was a retrospective consecutive case series of children aged < or =15 years given i.v. ketamine in the EDs of a university medical center and an affiliated county hospital over a 9-year period. A protocol for ketamine was used by treating physicians. Records were reviewed for adverse effects, indication, dosing, adjunctive drugs, inadequate sedation, and time to release. Results were contrasted with previously reported data for the i.m. route. RESULTS: During the study period i.v. ketamine was administered 156 times, primarily for laceration repair and fracture reduction. Transient apnea and respiratory depression occurred in one patient each; both were quickly identified and were without sequelae. Laryngospasm or aspiration was not noted in any children. There were 6 children with emesis and 2 with mild agitation during recovery. The median time from initial dose to ED release was 103 minutes (25th to 75th percentiles 76 to 146 minutes). The i.v. and i.m. routes were comparable in terms of adverse effects, inadequate sedation, and time to release. CONCLUSION: I.v. ketamine can be administered safely by EPs to facilitate pediatric procedures when used in a defined protocol. The sedation characteristics of the i.v. and i.m. routes appear comparable. PMID- 9862588 TI - A comparison of demand-valve and bag-valve ventilations in a swine pneumothorax model. AB - OBJECTIVE: Two means of delivering artificial ventilation readily available to out-of-hospital personnel are the bag-valve (BV) and the O2-powered demand-valve (OPDV). However, use of the OPDV has been limited because of concerns that it may worsen an underlying pneumothorax. This study compared the changes in size of pneumothorax in swine ventilated with the 2 devices. METHODS: Three swine were anesthetized, intubated, and instrumented with a femoral arterial line and a pediatric Swan-Ganz catheter. A chest tube was placed, the chest was opened, and the lung parenchyma was visualized. The lung was disrupted by a single stab with a #10 scalpel; the chest was then sealed; and a pneumothorax was created by injecting 30 mL of air through the chest tube. The animals were ventilated by 12 emergency medical technicians using either BV or OPDV. After 10 minutes of ventilation, the pneumothorax volume was measured. RESULTS: When comparing final pneumothorax volumes after 10 minutes of ventilation with the 2 devices, there was no significant difference (mean +/- SD = 40.8 +/- 28.2 mL vs 52.3 +/- 23.1 mL, p = 0.286). CONCLUSION: There is no difference in final pneumothorax volumes after OPDV or BV ventilation. PMID- 9862589 TI - How often do women in the emergency department without intimate violence injuries return with such injuries? AB - OBJECTIVE: To determine the rate at which a group of women visiting the ED for reasons other than intimate violence return to the ED at a later time for intimate-violence-related injuries. METHODS: Retrospective cohort study of a group of women with intimate-violence-related injuries on an index visit and a matched comparison group. Return visit rates to the ED for intimate violence injuries over the next 5 years were then compared. RESULTS: The 95 women in each group were followed an average of 57 months. The return rates in the positive index case group and matched comparison group, respectively, for any reason were 74.9% vs 77.9% (p = 0.463) and for intimate violence injuries were 29.5% vs 18.9% (p = 0.118). CONCLUSION: Women in the ED without intimate violence injuries often return to the ED later with such injuries. This suggests the ED may play an important role in identifying women at risk for future intimate-violence-related injury. PMID- 9862590 TI - A method for assessing emergency department performance using patient outcomes. AB - OBJECTIVE: To determine the rates of correct patient disposition after an ED evaluation. METHODS: In a university pediatric hospital, a 25% random sample of ED patients for 4 consecutive months was reviewed, after exclusion of minor injuries and patients triaged to the nonurgent clinic. Patients were categorized into one of 4 outcomes on the basis of inpatient resource use: appropriate admission, inappropriate admission, appropriate release, or inappropriate release. A 10% random sample of released patients was contacted by telephone to detect patients who sought care elsewhere after ED release. RESULTS: 642 of 2,682 ED patients (23.9%) were admitted; 159 (24.7%) were inappropriately admitted, and 26 (1.3%) were inappropriately released. The correct identification of the need for hospitalization (sensitivity) was 94.9%, and for release (specificity) 92.7%. Overall, the correct classification rate was 93.1%. Inappropriate admissions were associated with diagnoses of trauma, seizures, and burns. CONCLUSION: Inappropriate admissions occur at a substantial rate and occur more commonly than inappropriate releases. The correct disposition of patients is a practical and meaningful outcome-based measure of the quality of ED care. This methodology is suitable for use in other EDs. PMID- 9862591 TI - Carbon monoxide mass exposure in a pediatric population. AB - OBJECTIVES: To describe the outcomes of a mass carbon monoxide (CO) intoxication, and to calculate the CO half-life in a pediatric school-aged population. METHODS: A retrospective chart review was performed based on Regional Poison Center database information, hospital laboratory data, and medical records of the pediatric patients who sought care at one of 3 St. Louis area hospitals, after exposure to high levels of CO. Exposures occurred on January 5, 1996, after evidence of a CO leak was discovered at an area elementary school. Charts were reviewed for major demographics, symptoms reported, carboxyhemoglobin (COHb) levels and times, and level of effect. RESULTS: Information about 177 (35%) of the 504 children in attendance at school that day was available. Mean age was 8.7 +/- 1.8 years (range 4-12 years). Symptoms were present in 155 (88%) of the 177 children for whom data were available. Initial COHb levels were obtained for 147 (83.1%) of the 177 children. First mean COHb level was 7.0% (95% CI = 6.6-7.5%). Second COHb level was obtained for 26 children with a mean of 2.7% (95% CI = 2.2 3.2%). Calculated half-life of COHb, on 100% O2 at 1 atm, was 44.0 minutes (95% CI = 39.6-48.2 minutes). CONCLUSION: Some children had symptoms at COHb levels that traditionally have been considered nontoxic. The elimination of COHb was found to be more rapid in this population of children than reported in other studies. PMID- 9862592 TI - Follow-up compliance in febrile children: a comparison of two systems. AB - OBJECTIVES: Follow-up compliance is critical in febrile children because they may harbor unrecognized life-threatening illnesses. This study compares follow-up rates between 2 systems: Wilford Hall Medical Center (WHMC), with preset appointments after ED release, and free medical care; and Fairfax Hospital (FFX), where parents must arrange follow-up appointments after ED release, and are responsible for payment for their follow-up visits. The study also investigated factors associated with follow-up compliance. METHODS: This was a prospective, observational study of febrile children seen in 2 EDs with different systems for patient follow-up. From ED records and parental phone calls, diagnosis, follow-up compliance, and demographics were collected. Data were analyzed using logistic regression and chi2. RESULTS: 423 children met entrance criteria, and 330 parents were successfully contacted after the child's ED release (146 from WHMC; 184 from FFX). The WHMC children were more likely to comply with follow-up than were the children in the FFX system (92% vs 67% follow-up, odds ratio 2.5, 95% CI 1.1 5.3). Other factors associated with noncompliance with recommended follow-up were: Hispanic ethnicity, non-English-speaking parents, and follow-up suggested for >24 hours after ED release. For FFX, self-pay, lack of a follow-up physician, parents' dissatisfaction with the ED medical care, and diagnosis of otitis media were also significant factors found associated with noncompliance. CONCLUSION: Febrile children evaluated in a medical system with prearranged follow-up appointments and free medical care are more likely to comply with recommended follow-up than are those evaluated in a system where payment and appointments are the responsibility of the parents. Efforts should be made to improve follow-up compliance by modeling the WHMC system. PMID- 9862593 TI - Paramedic judgment of the need for trauma team activation for pediatric patients. AB - OBJECTIVE: To determine the value of paramedic judgment in determining the need for trauma team activation (TA) for pediatric blunt trauma patients. METHODS: A prospective, observational study was conducted at the ED of Children's Hospital Medical Center of Akron between July 12, 1996, and February 28, 1997, in cooperation with Akron Fire Department emergency medical technician-paramedics (EMT-Ps). The ED provides on-line and off-line medical control for pediatric transports. Patients with minor or no identifiable injuries are released at the scene with the instructions to see a physician. The remainder are transported to the ED. The decision for TTA is based on ED trauma protocols as well as emergency physician judgment of injury severity in combination with the judgment of the treating paramedic. During the study, EMT-Ps were asked (before physician input) whether, based solely on their judgment, a patient needed TTA. Patients 0-14 years old who were involved in motor vehicle crashes, bike crashes, or falls from a height of >10 feet were included in the study. TTA was defined as necessary if the patient was admitted to the intensive care unit (ICU) or operating room (OR) for nonorthopedic surgical procedures. Out-of-hospital, ED, and hospital records were reviewed. Coroners' records as well as medical records of all trauma admissions during the study period were reviewed to ensure that the patients released at the scene were not mistriaged. RESULTS: One hundred ninety-two patients met study criteria. Eighty-five patients (44%) were transported to the ED, of whom 12 had TTA. EMT-Ps requested TTA for 10 of these patients, and 2 patients had TTA per ED trauma protocol. Two of the patients who were judged by EMT-Ps to need TTA were admitted to the ICU/OR, and neither of the patients identified by ED trauma protocol to require TTA were admitted to the ICU/OR. Two initially stable patients who did not have TTA deteriorated after arrival to the ED. Both were admitted to the ICU. The sensitivity and specificity of paramedic judgment of the need for TTA for pediatric blunt trauma patients were 50% (95% CI 9.2-90.8) and 87.7% (95% CI 78.0-93.6), respectively. The positive and negative predictive values were 16.7% (95% CI 2.9-49.1) and 97.3% (95% CI 89.6-99.5). None of the patients released at the scene was mistriaged based on the review of the coroners' and trauma admission records. CONCLUSION: Results of this investigation indicate that a small percentage of pediatric blunt trauma patients require TTA. EMT-P judgment alone of the need for TTA for pediatric blunt trauma patients is not sufficiently sensitive to be of clinical use. The low sensitivity is explained by the deterioration in the clinical condition of 2 initially stable patients. The paramedic disposition decisions from the scene were always accurate. Nontransport by emergency medical services (EMS) may be acceptable in some uninjured pediatric trauma patients. Injured pediatric trauma patients who appear to be stable may deteriorate shortly after injury. However, if a pediatric patient appears injured, transport from the scene and examination by a trauma specialist are needed. Finally, the role of paramedic judgment must be further defined by larger studies with urban, rural, and suburban EMS systems before it can be used as a sole predictor of TTA. PMID- 9862594 TI - Determination of the minimal clinically significant difference on a patient visual analog satisfaction scale. AB - OBJECTIVE: To determine the minimal clinically significant difference (MCSD) on a visual analog patient satisfaction scale. METHODS: The authors prospectively collected patient satisfaction evaluations during a clinical trial assessing the effect of introducing personal television sets on overall patient satisfaction from their ED encounters. Patient satisfaction was assessed with 2 scales: a 100 mm visual analog scale (VAS) (0 = least satisfied, 100 = most satisfied) and a 7 point categorical scale ("terrible," "mostly dissatisfied," "mixed," "partially satisfied," "mostly satisfied," "pleased," and "delighted"). The differences between the mean VAS scores of "delighted" and "pleased" patients, and between "pleased" and "mostly satisfied" patients were used to determine the MCSD on the VAS. Reliability of each of the scales was determined. RESULTS: 181 patients were evaluated. Mean age was 41 years; 59% were female. On a subset of 19 patients, the VAS yielded an interobserver correlation of 0.93. The kappa measurement of agreement on the categorical scale was 0.77. The mean difference between "delighted" and "pleased" patient VAS satisfaction scores was 6.8 mm (95% CI, 1.3 12.3 mm). The mean difference between "pleased" and "mostly satisfied" patient VAS satisfaction scores was 10.7 mm (95% CI, 5.5-15.8 mm). CONCLUSION: The MCSD in patient satisfaction scores measured with a 100-mm VAS was approximately 7-11 mm. Future studies evaluating differences in patient satisfaction should be designed to detect this difference. PMID- 9862595 TI - Application of tissue adhesives: rapid attainment of proficiency. Stony Brook Octylcyanoacrylate Study Group. AB - OBJECTIVE: To evaluate the 3-month cosmetic outcome following laceration repair with a new tissue adhesive, 2-octylcyanoacrylate, as a function of physician experience with this tissue adhesive. METHODS: The authors prospectively enrolled consecutive patients >1 year of age with non-bite, non-crush-induced lacerations who presented <6 hours after injury and were treated with 2-octylcyanoacrylate. Structured closed-question data sheets were completed at the time of laceration repair and at 3-month follow-up. Long-term cosmetic appearance (>3 months) was assessed by patients using a 100-mm visual analog scale. The cosmetic outcomes were evaluated as a function of the physician application using ANOVA or chi2 tests, as appropriate. This study had 80% power to detect a 10-mm difference between the 2-octylcyanoacrylate and suture groups (alpha, 0.05). RESULTS: Seven physicians applied 2-octylcyanoacrylate to 63 patients; 61 patients received sutures. Patients were similar in the 2 groups (age, gender, race, history, and wound characteristics; p > 0.05 for all). At long-term follow-up, the cosmetic outcomes were similar in the 2-octylcyanoacrylate and suture groups according to patients (VAS 83.8 +/- 19.4 mm vs 82.5 +/- 17.6 mm; p = 0.72) and physicians (optimal score, 77% vs 80%; p = 0.67), and independent of physician experience with the 2-octylcyanoacrylate. One wound developed an infection and one wound necessitated reclosure due to dehiscence. Neither occurred with the first application. CONCLUSIONS: The 3-month cosmetic appearance of wounds treated with 2-octylcyanoacrylate is equivalent to that with sutures and does not improve as physicians become more experienced with use of this tissue adhesive. These data suggest that physicians can develop competence in application of tissue adhesives with a brief training period. PMID- 9862596 TI - Clinical pearls: Rash. PMID- 9862597 TI - Apoptosis. AB - Apoptosis is a process of cell suicide, the mechanisms of which are encoded in the genomes of all higher eukaryotes. The mechanisms involved in apoptosis suggest that the process is based on a viral defense originally developed in primitive multicelled eukaryotes and that the fundamental execution platform of the process involves 1) inhibition of protein synthesis at the level of translation initiation, 2) proteolysis specifically involving degradation of DNA repair mechanisms, and 3) polynucleotide degradation. In mammals this execution platform is regulated by a complex molecular signaling system that includes feedback mechanisms tending toward activation of all elements of the execution platform if only one element is initially engaged. Tissue ischemia and reperfusion activate elements of the apoptosis system, which thus represents a therapeutic target for emerging treatment approaches to preserve cellular integrity in critical organs such as the heart and brain. PMID- 9862598 TI - Statistical methodology: VII. Q-methodology, a structural analytic approach to medical subjectivity. AB - Q-methodology is a relatively unknown tool to medical researchers, that uses a mixed quantitative/qualitative statistical technique known as by-person factor analysis to study subjectivity. Q-methodology allows the grouping of individuals according to their subjective feelings about a topic, without requiring preconceived ideas regarding the structure of those subjective feelings. Q methodology is demonstrated using a study in which attitudes of emergency medicine residents toward computer education are explored systematically and classified in terms of statistically distinct factor viewpoints. In this example, Q-methodology identifies 4 attitude groups, as related to computers and computer education: 1) interested, eager to learn; 2) frustrated and interested, but with reservations; 3) interested mainly in benefits, willing to expend minimal effort; and 4) knowledgeable, independent learner. Q-methodology can be used to determine the structure of attitudes on a subjective topic, often yielding new insights. PMID- 9862599 TI - Spinal cord emergencies: false reassurance from reflexes. AB - Emergency physicians need to understand the potential for false reassurance in the interpretation of reflex examination data. Neurologic consultation should be sought when classic signs are lacking, but other evidence causes suspicion. Changes in teaching emphasis and acute practice are needed, since the stakes may be high and time is of the essence. We have responded to the insights gained from this study by augmenting in-service and continuing medical education teaching and by implementing guidelines to assist EPs. We have emphasized the importance of spotlighting high-risk patients, as exemplified above, and of taking advantage of neurologic or neurosurgical consultation. Where in-person consultation is less available, the use of guidelines and remote consultation should be able to help direct further examination, diagnostic formulation, and the need for imaging decisions. Given the potential for severe negative outcome if spinal emergencies are not optimally managed, we must give the teaching of these issues high priority. PMID- 9862601 TI - A comparison study of body dysmorphic disorder and obsessive-compulsive disorder. AB - BACKGROUND: Body dysmorphic disorder, a preoccupation with an imagined or slight defect in appearance, is classified as a somatoform disorder, but has been hypothesized to be related to obsessive-compulsive disorder. A reflection of this hypothesis, body dysmorphic disorder is included in the Yale-Brown Obsessive Compulsive Scale symptom checklist, and its transfer to the anxiety disorders' section was considered for DSM-IV. However, the similarities and differences between body dysmorphic disorder and obsessive-compulsive disorder have received little investigation. METHOD: We compared patients with DSM-IV body dysmorphic disorder (N = 53), obsessive-compulsive disorder (N = 53), or both disorders (N = 33) in terms of demographic features, clinical features, comorbidity, and family history. We also assessed the rate of body dysmorphic disorder among 62 of these subjects initially diagnosed with obsessive-compulsive disorder. RESULTS: 14.5% (9 of 62) of subjects initially diagnosed with obsessive-compulsive disorder had comorbid body dysmorphic disorder. The 2 disorders did not differ significantly in terms of sex ratio; most other demographic, course, and impairment variables; illness severity; or lifetime frequency of most associated disorders in probands or first-degree relatives. However, subjects with body dysmorphic disorder were less likely to be married and more likely to have had suicidal ideation or made a suicide attempt because of their disorder. They also had an earlier onset of major depression and higher lifetime rates of major depression, social phobia, and psychotic disorder diagnoses, as well as higher rates of substance use disorders in first-degree relatives. CONCLUSION: Body dysmorphic disorder appears to be relatively common among patients with obsessive-compulsive disorder. While the 2 disorders have many similarities, they also have some differences and should be differentiated in clinical and research settings. PMID- 9862600 TI - Getting stoned without inhaling: anandamide is the brain's natural marijuana. PMID- 9862602 TI - Rage attacks in children and adolescents with Tourette's disorder: a pilot study. AB - BACKGROUND: Sudden, explosive episodes of rage occur in a significant number of clinically referred children with Tourette's disorder and cause considerable psychosocial morbidity. The etiology of these symptoms is unknown. We conducted a pilot study of 12 consecutive children with Tourette's disorder and rage attacks to determine whether comorbidity of Tourette's-associated disorders is related to these symptoms. METHOD: Twelve consecutive children with Tourette's disorder who presented with rage attacks were evaluated, including 2 females and 10 males. Tourette's disorder diagnosis, presence of comorbid disorders, and tic severity were assessed using DSM-IV diagnostic criteria and standardized rating scales. RESULTS: All 12 children met diagnostic criteria for Tourette's disorder, obsessive-compulsive disorder (OCD), and attention-deficit/hyperactivity disorder (ADHD). Two children were also diagnosed with comorbid oppositional defiant disorder, and 4 children were diagnosed with comorbid conduct disorder. None of the subjects met diagnostic criteria for a mood disorder. All subjects had only mild tic severity. CONCLUSION: The clinical phenomenon of rage attacks in children with Tourette's disorder resembles intermittent explosive disorder and may reflect specific underlying neurologic disturbances. This pilot study suggests that rage attacks in Tourette's disorder may be related to the presence of comorbid disorders. PMID- 9862603 TI - Paroxetine treatment of episodic rages associated with Tourette's disorder. AB - BACKGROUND: Episodic rages have been estimated to occur in as many as 30% of patients with Tourette's syndrome (Tourette's disorder), but their treatment has never been systematically investigated. We report on the results of an open-label pilot study using paroxetine for the treatment of Tourette's disorder-associated rage episodes. METHOD: Forty-five Tourette's/rage patients (DSM-IV) were treated with paroxetine, specifically to control their rages. Other symptoms such as tics, attention-deficit/hyperactivity disorder (ADHD) and obsessive-compulsive disorder (OCD) were not targeted by this study. Treatment was deemed to be therapeutic when rage symptoms were diminished by 75% or more by patient report and were diminished in frequency by at least 1 point on a 4-point scale devised by the authors. RESULTS: After 8 weeks on paroxetine treatment, 29 patients (76% of those who completed the study) reported that rages were significantly diminished or completely absent. Nine patients reported no significant change in rages. Seven patients did not complete the study (3 because of side effects and 4 whose rage frequency increased). The mean dose of paroxetine was 33 mg/day; minimum effective dose was 15 mg/day. CONCLUSION: We were unable to determine any factors that significantly altered the efficacy of paroxetine for treatment of Tourette's disorder-associated rage episodes. The great majority (87%) of the patients had both ADHD and OCD in addition to Tourette's disorder. The age, sex, and concomitant use of other medications revealed no significant differences in treatment outcome. The results suggest that paroxetine may have an important role in the clinical treatment of episodic rages in Tourette's disorder patients. PMID- 9862604 TI - Switching from clozapine to olanzapine in treatment-refractory schizophrenia: safety, clinical efficacy, and predictors of response. AB - BACKGROUND: In our experience, many of our schizophrenic patients treated with clozapine request the newer atypical antipsychotic agents in order to eliminate the weekly blood monitoring. However, there are few guidelines available to clinicians interested in switching patients successfully treated with clozapine to olanzapine. METHOD: The goal of this study was to collect preliminary data on the safety, clinical effectiveness, and predictors of response of switching clozapine patients to olanzapine. In an open trial, 19 patients receiving clozapine were switched to olanzapine. RESULTS: Eight (42%) of 19 patients were considered responders. Seven patients decompensated seriously enough to require hospitalization. All 7 of these patients were restabilized on clozapine treatment in the hospital, and olanzapine was discontinued. In an additional 4 patients, clinical status worsened, and clozapine doses were titrated upwards and olanzapine was slowly discontinued. Overall, mean total Brief Psychiatric Rating Scale (BPRS) scores increased significantly from baseline to final assessment (p = .02). Responders had been treated for a significantly shorter period of time with clozapine prior to the switch compared to nonresponders (p = .04) and were receiving a lower dose of clozapine (p = .05). The final olanzapine dose did not differ between responders and nonresponders. All responders have remained on olanzapine treatment and are stable. CONCLUSION: In this open trial, the crossover from clozapine to olanzapine was generally well tolerated and resulted in a successful transition for 8 of the 19 patients. However, mean scores on the total BPRS and negative symptom and depressive symptom subscales significantly increased. Caution must be taken in determining which patients may benefit from the switch to olanzapine because of the risk of decompensation and hospitalization. Because this was an open trial, these findings require replication in a controlled trial. PMID- 9862605 TI - The treatment of chronic depression, part 1: study design and rationale for evaluating the comparative efficacy of sertraline and imipramine as acute, crossover, continuation, and maintenance phase therapies. AB - BACKGROUND: Chronic depressions are common, disabling, and undertreated, and prior chronicity predicts future chronicity. However, few studies directly inform the acute or maintenance phase treatments of chronic depressions and even less is known about the effects of treatment on psychosocial functioning. METHOD: We describe the design and rationale for 2 parallel double-blind, randomized, multicenter acute and maintenance phase treatment trials. One focused on DSM-III R major depression currently in a chronic (> or = 2 years) major depressive episode, the other on DSM-III-R major depression with concurrent DSM-III-R dysthymia ("double depression"). RESULTS: Considering the critical knowledge deficits, we designed a 12-week acute phase safety and efficacy trial of sertraline versus imipramine, followed by a 16-week continuation treatment phase for subjects with a satisfactory therapeutic response. Patients receiving sertraline who successfully completed the continuation phase entered a 76-week maintenance trial to compare sertraline with placebo; those taking imipramine continued without a placebo substitution. As part of the acute trial, subjects completing but failing to respond to the initial 12-week acute phase medication were crossed over (double-blind) to the alternative medication for a 12-week acute phase trial. We obtained naturalistic follow-up data (up to 18 months) for subjects exiting the protocol at any time. CONCLUSION: Multiphase protocols for chronic depression can test efficacy by randomized contrasts as well as shed light on key clinical issues such as the degree of response or attrition expected at particular times in a trial or the preferred medication sequence in a potential multistep treatment program. PMID- 9862606 TI - The treatment of chronic depression, part 2: a double-blind, randomized trial of sertraline and imipramine. AB - BACKGROUND: Chronic depression appears to be a common, frequently disabling illness that is often inadequately treated. Unlike episodic depressions with shorter illness duration, neither acute nor long-term treatment approaches for chronic depression have been well studied. METHOD: 635 outpatients at 12 sites who met DSM-III-R criteria for chronic major depression or double depression were randomly assigned to 12 weeks of double-blind treatment with either sertraline (in daily doses of 50-200 mg) or imipramine (in daily doses of 50-300 mg). Efficacy and safety were assessed either weekly or every 2 weeks during the 12 weeks of acute treatment. RESULTS: Despite high rates of chronicity (mean duration of major depression = 8.9+/-9.1 years; mean duration of dysthymia = 23+/ 13 years) and high rates of comorbidity, 52% of patients achieved a satisfactory therapeutic response to sertraline or imipramine (by a conservative, intent-to treat analysis). Approximately 21% of the patients who had achieved a therapeutic response at week 12 had not done so at week 8, confirming the longer time to response in depressions with high chronicity. Patients treated with sertraline reported significantly fewer adverse events and were significantly less likely to discontinue treatment due to side effects than imipramine-treated patients (6.3% vs. 12.0%). CONCLUSION: These results indicate that patients suffering from depression with high chronicity can achieve a good therapeutic response to acute treatment with either sertraline or imipramine, although sertraline is better tolerated. PMID- 9862607 TI - The treatment of chronic depression, part 3: psychosocial functioning before and after treatment with sertraline or imipramine. AB - BACKGROUND: Previous research has suggested that depressed patients, and particularly chronically depressed patients, have significant impairments in many areas of their lives. While previous studies suggested that these "psychosocial" impairments improve following pharmacologic treatment, no large scale definitive study using multiple measures of psychosocial functioning has been reported. METHOD: We assessed multiple domains of psychosocial functioning using interviewer-rated and self-report measures within the context of a 12-week acute treatment trial of sertraline and imipramine for patients with chronic depression (double depression and chronic major depression). We also compared the psychosocial functioning data of this sample before and after treatment with normative data available from published community samples. RESULTS: Chronically depressed patients manifested severe impairments in psychosocial functioning at baseline. After treatment with sertraline or imipramine, psychosocial functioning improved significantly. Significant improvements appeared relatively early in treatment (week 4). Despite these highly significant improvements in functioning during acute treatment, the study sample as a whole did not achieve levels of psychosocial functioning comparable to a comparator nondepressed community sample. However, patients who reached full symptomatic response (remission) during acute treatment did have levels of psychosocial functioning in most areas at endpoint that approached or equaled those of community samples. CONCLUSION: These results indicate that successful antidepressant treatment with sertraline or imipramine can alleviate the severe psychosocial impairments found in chronic depression. PMID- 9862608 TI - Conflicting and inconsistent dosing advice for clonidine in Tourette's syndrome. PMID- 9862609 TI - Manic and psychotic symptoms following risperidone withdrawal in a schizophrenic patient. PMID- 9862610 TI - Treatment of venlafaxine discontinuation symptoms with ondansetron. PMID- 9862611 TI - Fulminant hepatic failure from acetaminophen in an anorexic patient treated with carbamazepine. PMID- 9862612 TI - Movement disorders and psychotic symptoms treated with pyridoxine: a case report. PMID- 9862613 TI - Risperidone monotherapy for psychotic depression unresponsive to other treatments. PMID- 9862614 TI - The naturalistic course of pharmacologic treatment of children with maniclike symptoms: a systematic chart review. AB - OBJECTIVE: To assess the effectiveness of mood stabilizers in treating maniclike symptoms in children. METHOD: Subjects were consecutively referred pediatric patients who, at initial intake, satisfied DSM-III-R criteria for mania on a structured diagnostic interview. We systematically reviewed their clinical records to assess (1) the course of maniclike symptoms and (2) all medications prescribed at each follow-up visit. Survival analysis was used to determine the effect of mood stabilizers and other medications on the course of maniclike symptoms. RESULTS: Of the 59 subjects meeting criteria for mania, 44 (75%) exhibited evidence of maniclike symptoms during follow-up. The occurrence of manic symptoms significantly predicted the subsequent prescription of mood stabilizers (rate ratio = 2.9, 95% confidence interval [CI] = 1.6 to 5.5), and use of mood stabilizers predicted decreases in manic symptoms (rate ratio = 4.9, 95% CI = 1.2 to 20.8). However, improvement was slow and associated with a substantial risk for relapse. CONCLUSION: Mood stabilizers were frequently used in children with maniclike symptoms, and their use was associated with significant improvement of maniclike symptoms, whereas use of antidepressant, antipsychotic, and stimulant medications was not. PMID- 9862615 TI - Sun safety: avoiding noonday sun, wearing protective clothing, and the use of sunscreen. PMID- 9862616 TI - Depression as a risk factor for cancer: renewing a debate on the psychobiology of disease. PMID- 9862618 TI - Days, weeks, or months: when to say how long? PMID- 9862617 TI - Quest for new and better colon cancer treatments picks up steam. PMID- 9862619 TI - Metaphorically speaking: DNA spoken here. PMID- 9862620 TI - Economically disadvantaged may survive cancer longer in Canada. PMID- 9862621 TI - Database provides window on applications of treatments. PMID- 9862622 TI - Scientists explore use of arsenic in therapy. PMID- 9862623 TI - Evaluating the efficacy of screening for recurrence of cancer. PMID- 9862624 TI - Sunscreen use, wearing clothes, and number of nevi in 6- to 7-year-old European children. European Organization for Research and Treatment of Cancer Melanoma Cooperative Group. AB - BACKGROUND: Previous epidemiologic studies have suggested that sunscreen use is associated with an increased risk of melanoma skin cancer. Because high nevi (mole) count in adults is a strong predictor of melanoma, we conducted a study examining the number of nevi in 6- to 7-year-old European children, according to their sunscreen use. METHODS: Whole-body and site-specific counts of nevi 2 mm or larger were performed in 631 children in their first year of primary school in four European cities. Independently, parents were interviewed regarding sun exposure, sunscreen use, and physical sun protection of their child. RESULTS: After adjustment for sun exposure and host characteristics (e.g., skin phototype, eye color), the relative risk for high nevus count on the trunk was 1.68 (95% confidence interval [CI] = 1.09-2.59) for the highest level of sunscreen use and 0.59 (95% CI = 0.36-0.97) for the highest level of wearing of clothes while in the sun. The sun protection factor had no effect on nevus counts despite a high median value of 17.4. Sunburn number was not associated with nevus count. The highest risk associated with sunscreen use was found among children who had never experienced sunburn. CONCLUSIONS: In white, European children, sunscreen use appears to be associated with development of nevi, probably because it allows longer sun exposures. Wearing clothes may be an effective way to prevent proliferation of nevi. Since a high nevus count is a strong predictor of melanoma, sunscreen use may be involved in melanoma occurrence because it may encourage recreational sun exposure. PMID- 9862626 TI - Chronically depressed mood and cancer risk in older persons. AB - BACKGROUND: Depression has been proposed as a predisposing factor for cancer, but prospective studies have been inconclusive. We examined whether a high level of depressive symptoms, present for a long time, is associated with increased risk of cancer in the elderly. METHODS: Data were obtained and analyzed from persons who lived in three communities (Massachusetts, Iowa, and Connecticut) of the Established Populations for Epidemiologic Studies of the Elderly, a prospective cohort study with a mean follow-up of 3.8 years that included 4825 persons (1708 men and 3117 women) aged 71 years and older. Chronically depressed mood was defined as present when the number of depressive symptoms exceeded specific cut points on the Center for Epidemiologic Studies-Depression scale at baseline (1988) and 3 and 6 years before baseline. New cases of cancer were identified from Medicare hospitalization records and death certificates. RESULTS: Of the 4825 persons studied, 146 (3.0%) were chronically depressed. The incidence rate of cancer was 30.5 per 1000 person-years for the 146 persons with chronic depression and 21.9 per 1000 person-years for the 4679 nonchronically depressed persons. After adjustment for age, sex, race, disability, hospital admissions, alcohol intake, and smoking, the hazard ratio for cancer associated with chronically depressed mood was 1.88 (95% confidence interval = 1.13-3.14). The excess risk of cancer associated with chronic depression was consistent for most types of cancer and was not specific to cigarette smokers. CONCLUSION: When present for at least 6 years, depression was associated with a generally increased risk of cancer. PMID- 9862625 TI - Construction and characterization of a triple-recombinant vaccinia virus encoding B7-1, interleukin 12, and a model tumor antigen. AB - BACKGROUND: Construction of recombinant viruses that can serve as vaccines for the treatment of experimental murine tumors has recently been achieved. The cooperative effects of immune system modulators, including cytokines such as interleukin 12 (IL-12) and costimulatory molecules such as B7-1, may be necessary for activation of cytotoxic T lymphocytes. Thus, we have explored the feasibility and the efficacy of inclusion of these immunomodulatory molecules in recombinant virus vaccines in an experimental antitumor model in mice that uses Escherichia coli beta-galactosidase as a target antigen. METHODS: We developed a "cassette" system in which three loci of the vaccinia virus genome were used for homologous recombination. A variety of recombinant vaccinia viruses were constructed, including one virus, vB7/beta/IL-12, that contains the following five transgenes: murine B7-1, murine IL-12 subunit p35, murine IL-12 subunit p40, E. coli lacZ (encodes beta-galactosidase, the model antigen), and E. coli gpt (xanthine guanine phosphoribosyltransferase, a selection gene). The effects of the recombinant viruses on lung metastases and survival were tested in animals that had been given an intravenous injection of beta-galactosidase-expressing murine colon carcinoma cells 3 days before they received the recombinant virus by intravenous inoculation. RESULTS: Expression of functional B7-1 and IL-12 by virally infected cells was demonstrated in vitro. Lung tumor nodules (i.e., metastases) were reduced in mice by more than 95% after treatment with the virus vB7/beta/IL-12; a further reduction in lung tumor nodules was observed when exogenous IL-12 was also given. Greatest survival of tumor-bearing mice was observed in those treated with viruses encoding beta-galactosidase and B7-1 plus exogenous IL-12. CONCLUSION: This study shows the feasibility of constructing vaccinia viruses that express tumor antigens and multiple immune cofactors to create unique immunologic microenvironments that can modulate immune responses to cancer. PMID- 9862628 TI - Cancer of the oral cavity and pharynx in nonsmokers who drink alcohol and in nondrinkers who smoke tobacco. PMID- 9862629 TI - Application of heavy metal and cytokine for differentiation-inducing therapy in acute promyelocytic leukemia. PMID- 9862627 TI - Immunizing patients with metastatic melanoma using recombinant adenoviruses encoding MART-1 or gp100 melanoma antigens. AB - BACKGROUND: The characterization of the genes encoding melanoma-associated antigens MART-1 or gp100, recognized by T cells, has opened new possibilities for the development of immunization strategies for patients with metastatic melanoma. With the use of recombinant adenoviruses expressing either MART-1 or gp100 to immunize patients with metastatic melanoma, we evaluated the safety, immunologic, and potential therapeutic aspects of these immunizations. METHODS: In phase I studies, 54 patients received escalating doses (between 10(7) and 10(11) plaque forming units) of recombinant adenovirus encoding either MART-1 or gp100 melanoma antigen administered either alone or followed by the administration of interleukin 2 (IL-2). The immunologic impact of these immunizations on the development of cellular and antibody reactivity was assayed. RESULTS: Recombinant adenoviruses expressing MART-1 or gp100 were safely administered. One of 16 patients with metastatic melanoma receiving the recombinant adenovirus MART-1 alone experienced a complete response. Other patients achieved objective responses, but they had received IL-2 along with an adenovirus, and their responses could be attributed to the cytokine. Immunologic assays showed no consistent immunization to the MART-1 or gp100 transgenes expressed by the recombinant adenoviruses. High levels of neutralizing antibody were found in the pretreatment sera of the patients. CONCLUSIONS: High doses of recombinant adenoviruses could be safely administered to cancer patients. High levels of neutralizing antibody present in patients' sera prior to treatment may have impaired the ability of these viruses to immunize patients against melanoma antigens. PMID- 9862630 TI - Re: Physical activity and breast cancer risk in a cohort of young women. PMID- 9862631 TI - Re: Relationship between topotecan systemic exposure and tumor response in human neuroblastoma xenografts. PMID- 9862632 TI - The definition of multiple system atrophy: a review of recent developments. PMID- 9862633 TI - SPARC: a signal of astrocytic neoplastic transformation and reactive response in human primary and xenograft gliomas. AB - In an attempt to identify genetic alterations occurring early in astrocytoma progression, we performed subtractive hybridization between astrocytoma and glioblastoma cDNA libraries. We identified secreted protein acidic and rich in cysteine (SPARC), a protein implicated in cell-matrix interactions, as a gene overexpressed early in progression. Northern blot and immunohistochemical analyses indicated that transcript and protein were both elevated in all tumor specimens (grades II-IV) examined when compared with levels in normal brain. The level of SPARC expression was found to be tumor-dependent rather than grade related. Immunohistochemically, SPARC protein was found to be overexpressed in 1) cells in the less cellularly dense regions within the tumor mass, 2) histomorphologically neoplastic-looking cells in adjacent normal brain at the tumor/brain interface, 3) neovessel endothelial cells in both the tumor and adjacent normal brain, and 4) reactive astrocytes in normal brain adjacent to tumor. Using a combination of DNA in situ hybridization and protein immunohistochemical analyses of the human/rat xenograft, SPARC expression was observed in the human glioma cells within the tumor mass, and in cells that invaded along vascular basement membranes and individually into the rat brain parenchyma, suggesting it may be an invasion-related gene. While it remains to be determined whether SPARC functionally contributes to tumor cell invasion, these data suggest that the early onset of increased SPARC expression, though complex, may serve as a signal indicative of neoplastic astrocytic transformation and reactive response to tumor-induced stress. PMID- 9862634 TI - Abeta associated neuropil changes: correlation with neuronal loss and dementia. AB - Although genetic studies clearly implicate beta-amyloid peptide (Abeta) as a pathogenic agent in Alzheimer disease (AD), it is puzzling that the total amount of Abeta immunoreactivity does not correlate closely with neuronal loss or degree of dementia. We hypothesized that Abeta deposits could vary in the extent to which they disrupt the neuropil, and that the degree to which this occurs might then correlate with the degree of dementia. We used 3 dimensional triple immunofluorescent confocal microscopy to examine the fine structural relationships between Abeta deposits and neurites in their vicinity. In non demented elderly, Abeta deposits were porous structures with numerous normal appearing processes coursing through them. In AD, dendrites within Abeta deposits, compared with dendrites in the surrounding neuropil, were likely to have decreased SMI32 immunoreactivity and increased Alz-50 immunoreactivity. We found that the degree to which Abeta deposits disrupt the neuropil, as assessed by local loss of SMI32 immunoreactivity, correlates closely with the amount of neuronal loss and with duration of dementia. These observations support the hypothesis that a subset of Abeta deposits contribute directly to neural system failure in AD. PMID- 9862635 TI - Extracellular neurofibrillary tangles are immunopositive for the 40 carboxy terminal sequence of beta-amyloid protein. AB - Neurofibrillary tangles (NFTs) form in a number of neurodegenerative disorders. In Alzheimer disease (AD), intracellular NFTs (iNFTs) develop along with extracellular beta-amyloid (Abeta) deposits. Reports on whether NFTs have Abeta associated with them are inconsistent. Here we study NFTs and their direct relationship with Abeta-like fragments in cases of AD, Down Syndrome, and the parkinsonism-dementia complex of Guam, using a panel of antibodies which recognize different epitopes of Abeta. In all diseases, as well as in the aged controls, the majority of extracellular NFTs (eNFTs) are stained with antibodies recognizing the 40 carboxy-terminal of Abeta, but not other epitopes. Such staining is morphologically distinguishable from the previously described Abeta positive 'tangle associated amyloid deposits' (TAADs), which surround some eNFTs, and are immunopositive for all epitopes of the Abeta molecule. Some iNFTs are immunoreactive with antibodies to the 42 carboxy-terminal epitope, and, to a lesser extent, with antibodies to midportions and more N-terminal epitopes of Abeta. These results may indicate a direct interaction between Abeta and NFTs, although secondary deposition or crossreactivity with other epitopes associated with NFTs cannot be ruled out. PMID- 9862636 TI - Chromosome 7 rearrangements in glioblastomas; loci adjacent to EGFR are independently amplified. AB - The first gene found to be amplified in human glioblastomas was EGFR at 7p12. More recently the MET gene at 7q31 was also reported amplified. We have studied chromosome 7 in a series of 47 glioblastomas by FISH, RFLP and microsatellite analysis. Four per cent (2/47) had 1 centromere, 26% (12/47) 2, 32% (15/47) 3, 4% (2/47) 4, and 34% (16/47) had subpopulations with variable numbers of chromosome 7 centromeres. In 25 of the 47 tumors (53%) the pattern of allelic imbalance observed at each informative locus was similar and in accord with the FISH data, indicating loss or gain of complete chromosome copies. In 32% of tumors (15/47) varying allelic imbalance was seen at different loci along the chromosome indicative of loss or gain of parts of chromosome 7 on a background of disomy, trisomy, tetrasomy, or polysomy. Amplification was studied in an extended series of 121 glioblastomas, and was seen at the 7p12 region in 47 tumors (39%). Forty two tumors showed amplification of EGFR and 12 of these had extensive amplicons including a number of adjacent loci, always involving only 1 allele. The amplicons of 5 tumors (11%) did not include EGFR, indicating that other unidentified genes in the region are targeted for amplification. Amplification of MET was not found. The findings show that copy number changes of chromosome 7 are common and that a number of genes may be targeted for amplification at 7p12 in glioblastomas. PMID- 9862637 TI - Synaptic density in the inner molecular layer of the hippocampal dentate gyrus in Alzheimer disease. AB - We examined the inner molecular layer (IML) of the hippocampal dentate gyrus for possible changes in synaptic density. Material was obtained from 9 individuals with Alzheimer disease (AD) and compared to samples obtained from 10 age-matched, postmortem-matched neurologically normal controls, employing standard ultrastructural techniques. Statistical analyses demonstrated a significant decline in synaptic numbers between controls and AD subjects. This decline was accompanied by a significant increase in apposition length and resulted in a significant correlation with the synaptic density. As the number of synapses declined, the apposition length increased. Assessment was also made of the granule cells density and the analyses showed a significant decline in the synapse to granule cell ratio in the AD group. This decline in the density of synaptic contacts in the IML reflects a more widespread decline in plasticity in AD and may be related to the memory problems associated with the disease. PMID- 9862638 TI - Monitoring the CNS pathology in aspartylglucosaminuria mice. AB - Aspartylglucosaminuria (AGU) is a recessively inherited lysosomal storage disorder caused by the deficiency of the aspartylglucosaminidase (AGA) enzyme. The hallmark of AGU is slowly progressing mental retardation but the progression of brain pathology has remained uncharacterized in humans. Here we describe the long-term follow-up of mice carrying a targeted AGU-mutation in both alleles. Immunohistochemistry, histology, electron microscopy, quantitative magnetic resonance imaging (MRI) and behavioral studies were carried out to evaluate the CNS affection of the disease during development. The lysosomal storage vacuoles of the AGA -/- mice were most evident in central brain regions where MRI also revealed signs of brain atrophy similar to that seen in the older human patients. By immunohistochemistry and MRI examinations, a subtle delay of myelination was observed in AGA -/- mice. The life span of the AGA -/- mice was not shortened. Similar to the slow clinical course observed in human patients, the AGA -/- mice have behavioral symptoms that emerge at older age. Thus, the AGU knock-out mice represent an accurate model for AGU, both histopathologically and phenotypically. PMID- 9862639 TI - Loss of the NF2 gene and merlin occur by the tumorlet stage of schwannoma development in neurofibromatosis 2. AB - Loss of the neurofibromatosis 2 (NF2) gene-encoded protein merlin is a universal finding in sporadic and NF2-associated schwannomas. Certain NF2 patients may develop numerous minute Schwann cell tumorlets of the spinal nerve roots in addition to larger, frank schwannomas and thereby provide an opportunity to investigate the timing of NF2 gene/merlin loss in Schwann cell tumorigenesis. We studied an NF2 patient with a germline NF2 gene frameshift mutation who had many Schwann cell tumorlets and schwannomas. Loss of heterozygosity studies of DNA from microdissected specimens showed allelic loss of the NF2 region of chromosome 22q in tumorlets as well as schwannomas. Immunohistochemistry further demonstrated loss of merlin expression in tumorlets as well as schwannomas, with intact expression in adjacent nerve. Thus, loss of both NF2 alleles and merlin occur early in Schwann cell tumorigenesis, before the tumorlet stage. The study of tumorlets and schwannomas in such patients may also provide an opportunity to elucidate mechanisms responsible for the subsequent growth of Schwann cell lesions into symptomatic tumors. PMID- 9862640 TI - Neuropathological and neuropsychological changes in "normal" aging: evidence for preclinical Alzheimer disease in cognitively normal individuals. AB - The presence of diffuse or primitive senile plaques in the neocortex of cognitively normal elderly at autopsy has been presumed to represent normal aging. Alternatively, these patients may have developed dementia and clinical Alzheimer disease (AD) if they had survived. In this setting, these patients could be subjects for cognitive or pharmacologic intervention to delay disease onset. We have thus followed a cohort of cognitively normal elderly subjects with a Clinical Dementia Rating (CDR) of 0 at autopsy. Thirty-one brains were examined at postmortem according to Consortium to Establish a Registry for Alzheimer Disease (CERAD) criteria and staged according to Braak. Ten patients were pathologically normal according to CERAD criteria (1a). Two of these patients were Braak Stage II. Seven very elderly subjects exhibited a few primitive neuritic plaques in the cortex and thus represented CERAD 1b. These individuals ranged in age from 85 to 105 years and were thus older than the CERAD la group that ranged in age from 72 to 93. Fourteen patients displayed Possible AD according to CERAD with ages ranging from 66 to 95. Three of these were Braak Stage I, 4 were Braak Stage II, and 7 were Braak Stage III. The Apolipoprotein E4 allele was over-represented in this possible AD group. Neuropsychological data were available on 12 individuals. In these 12 individuals, Possible AD at autopsy could be predicted by cognitive deficits in 1 or more areas including savings scores on memory testing and overall performance on some measures of frontal executive function. PMID- 9862641 TI - Effect of sorbitol dehydrogenase inhibition on experimental diabetic autonomic neuropathy. AB - The polyol pathway and its dependent biochemical pathways are thought to play a role in the pathogenesis of diabetic neuropathy. We have developed an animal model of diabetic autonomic neuropathy characterized by neuroaxonal dystrophy involving ileal mesenteric nerves and prevertebral sympathetic superior mesenteric ganglia (SMG) in chronic streptozocin-diabetic rats. Our previous studies have shown a salutary effect of aldose reductase inhibitors on experimental autonomic neuropathy, suggesting a role for the polyol pathway in its pathogenesis. In the current studies we have examined the effect of the sorbitol dehydrogenase inhibitor (SDI) CP-166,572, which interrupts the conversion of sorbitol to fructose (and reactions dependent on the second step of the polyol pathway) resulting in markedly increased levels of sorbitol in peripheral nerve. Fourteen weeks of treatment with CP-166,572 resulted in a dramatically increased frequency of neuroaxonal dystrophy in ileal mesenteric nerves and SMG. Although lesions developed prematurely and in greater numbers in SDI-treated diabetics than untreated diabetics did, their anatomic distribution and ultrastructural appearance were identical to that previously reported in long term untreated diabetics. CP-166,572 treatment did not produce neuroaxonal dystrophy in control animals despite the fact that sciatic nerve sorbitol levels were markedly increased, reaching the same levels as untreated diabetic animals. Treatment of diabetic rats for 14 weeks with the aldose reductase inhibitor zopolrestat resulted in a significant decrease in the frequency of neuroaxonal dystrophy compared with untreated diabetics. PMID- 9862643 TI - Curly fiber and tangle-like inclusions in the ependyma and choroid plexus--a pathogenetic relationship with the cortical Alzheimer-type changes? AB - The question of whether thread- and tangle-like inclusions of the choroid plexus (known as Biondi inclusions) are related to the cortical lesions in Alzheimer disease (AD) has been debated for almost a century, yet remains unanswered. Recently beta-amyloid protein was biochemically isolated from the plexus, indicating a possible pathogenetic relationship between the degenerative changes of the cerebral cortex and those of the plexus. The goal of the present study was to analyze whether or not a significant correlation exists between the occurrence of the cortical AD-type changes and those in the ependyma and choroid plexus. In 292 consecutive autopsy cases several cortical areas, the ependyma, and the choroid plexus were analyzed to look for AD-type changes and Biondi inclusions using histochemical staining techniques and immunohistochemistry. A semiquantitative analysis of the density of cortical AD-type changes showed that of the 292 cases, 63 had severe cortical changes, 23 moderate changes, and 142 discrete changes. In 64 cases no plaques or neurofibrillary tangles were found. The number of cases with thread- and tangle-like elements in the plexus and ependyma was more than 96% in the 3 groups with cortical AD-type lesions, but low in the group without AD-type cortical changes (19%). The pathological argyrophilic filaments accumulating in the ependymal layer and plexus had histochemical properties of amyloid and were immunoreactive with antibodies to P component, ubiquitin, fibronectin and Tau protein. They did not react with antibodies to neurofilament proteins. Ultrastructurally, they consisted of densely packed straight and paired helical filaments and closely resembled neurofibrillary tangles and neuropil threads. The highly significant correlation (chi2, p = 0.001; R = 0.85) between the occurrence of AD-type changes in the cortex and those in ependyma and plexus suggests a pathogenetic relationship. PMID- 9862642 TI - Intraneuronal ApoE in human visual cortical areas reflects the staging of Alzheimer disease pathology. AB - Alzheimer disease (AD) is marked by progressive loss of cortical neurons with associated cognitive decline. Multiple genetic and environmental factors likely contribute to this progressive loss. Such genetic factors include the polymorphic locus (APOE) that encodes apolipoprotein E (apoE). In order to investigate a possible correspondence between cellular localization of apoE and the neuropathology of AD, we examined the distribution of apoE-immunoreactive neurons in visual cortical areas with different apparent susceptibility to AD neuropathology (areas 17-primary sensory, 18-secondary sensory, and inferior temporal-association cortex) at different stages of AD pathology as described by Braak and Braak. We found that intraneuronal apoE was present at all these stages, however, only in visual cortical regions known to be vulnerable to AD. In the late stages, the laminar distribution of apoE-immunoreactivity matched the distribution of other markers of AD pathology, especially modified tau. These data support previous findings that intraneuronal apoE in neocortex is common in aged, nondemented controls and demonstrate that it may be more common in regions at risk for AD pathology. Thus, intraneuronal accumulation of apoE may be an attribute of cortical neurons that are more vulnerable to age-related injury with the presence of apoE antedating the classical indices of late-onset AD pathology. PMID- 9862644 TI - Formation of DNA adducts of the food-derived mutagen 2-amino-9H-pyrido-[2,3 b]indole (A(alpha)C) and bioassay of mammary gland carcinogenicity in Sprague Dawley rats. AB - 2-amino-9H-pyrido[2,3-b]indole (AalphaC) is a heterocyclic amine found at relatively high concentrations in barbecued or grilled meats. In the current study, the mammary gland carcinogenicity of AalphaC was examined in female Sprague-Dawley rats given 10 doses of AalphaC (75 mg/kg, orally, once per day starting at 43 days of age) and placed on a defined high-fat diet (23.5% corn oil), a strong promotional factor for rat mammary gland carcinogenesis. Within 1 year, one out of 20 rats dosed with AalphaC developed a tubulopapillary carcinoma, indicating that the bioassay was largely negative. As DNA adduct formation is considered to play a role in carcinogenesis, AalphaC-DNA adduct levels were measured in the mammary gland and other tissues by the 32P postlabelling method. Under intensification conditions, one major adduct and up to three minor adducts were detected in isolated mammary gland epithelial cells and other tissues (liver, stomach, small intestine, colon and kidney) of AalphaC treated rats; the adduct patterns were similar in all tissues examined. The major adduct, comprising 60-100% of total DNA adduct levels in tissues, was chromatographically identical to the principal adduct found in 3'-dGp-AalphaC (synthesized by reacting 3'-phospho-2'-deoxyguanosine (3'-dGp) with N-acetoxy AalphaC). Of the tissues examined, the highest AalphaC-DNA adduct levels were found in the liver. In male rats given a single dose of AalphaC (75 mg/kg, orally, 3 hr prior to necropsy), no AalphaC-DNA adducts were detected in extrahepatic tissues. In female rats given a single dose or 12 daily doses of AalphaC, hepatic DNA adduct levels were at least 12-13-fold higher than those in any other tissue. Mean total AalphaC-DNA adduct levels in mammary gland epithelial cells and liver from female rats given multiple doses of AalphaC were 3.5 and 50.7 (RAL x 10(7)), respectively. Although factors in addition to DNA adduct formation are likely to play a role in mammary gland carcinogenesis, the results suggest that the weak mammary gland carcinogenicity of AalphaC may in part be associated with low AalphaC-DNA adduct levels in the mammary gland epithelium. PMID- 9862645 TI - Protective effects of green tea on hepatotoxicity, oxidative DNA damage and cell proliferation in the rat liver induced by repeated oral administration of 2 nitropropane. AB - To evaluate the benefit of green tea in mitigating hazards caused by repeated exposure of 2-nitropropane (2NP), we examined the effects of the tea on toxic indices, oxidative DNA damage and cell proliferation in the liver of 2NP-treated rats. Male Fischer 344 rats were administered, by gastric intubation, a total of six doses of 60 mg/kg 2NP(L), or alternatively two doses of 90 mg/kg and then four doses of 120 mg/kg 2NP(H) during 2 weeks. Green tea infusion was given to the rats as drinking water 1 week before the 2NP treatments and throughout the experiment. Significant elevation of hepatotoxic indices was evident in the 2NP(H)-treated group, such as an increase of serum glutamic-oxaloacetic transaminase (GOT) activity and of hepatic lipid peroxidation, together with a decrease in hepatic glycogen and serum triglyceride, and degenerative changes in the hepatocytes. A dose-related increase was observed in oxidative DNA damage and cell proliferation in the liver. Green tea effectively inhibited all of above changes induced by 2NP treatment, suggesting that tea intake may be effective for preventing the hepatic injuries after chronic exposure to 2NP. PMID- 9862646 TI - Mechanism of inhibition of tannic acid and related compounds on the growth of intestinal bacteria. AB - Tannic acid, propyl gallate and methyl gallate, but not gallic acid, were found to be inhibitory to the growth of intestinal bacteria Bacteroides fragilis ATCC 25285, Clostridium clostridiiforme ATCC 25537, C. perfringens ATCC 13124, C. paraputrificum ATCC 25780, Escherichia coli ATCC 25922, Enterobacter cloacae ATCC 13047, Salmonella typhimurium TA98 and S. typhimurium YG1041 at 100-1000 microg/ml in culture broth. Neither Bifidobacterium infantis ATCC 15697 nor Lactobacillus acidophilus ATCC 4356 was inhibited by any of the above compounds up to 500 microg/ml. Tannic acid has a much greater relative binding efficiency to iron than propyl gallate, methyl gallate or gallic acid. The inhibitory effect of tannic acid to the growth of intestinal bacteria may be due to the strong iron binding capacity of tannic acid; whereas the effect of propyl gallate and methyl gallate probably occurs by a different mechanism. The growth of E. coli was restored by the addition of iron to the medium after the precipitate caused by tannic acid was removed. Neither B. infantis nor L. acidophilus require iron for growth. This probably contributes to their resistance to tannic acid. Because tannins are abundant in the human diet, tannins may affect the growth of some intestinal bacteria and thus may have an impact on human health. PMID- 9862647 TI - Safety evaluation and fluorine concentration of Pu'er brick tea and Bianxiao brick tea. AB - Pu'er brick tea and Bianxiao brick tea are both compressed types of tea. Fluorine analysis was carried out on samples of Pu'er brick tea produced at different times in Yunnan Province and on samples of Bianxiao brick tea made in Hunan and Sichuan Province for supply especially to minority ethnic groups in border areas of China. The levels of water-soluble and water-insoluble fluorine were measured in the tea samples using an ion-specific electrode potentiometer. The concentration of water-soluble fluorine was much greater in Bianxiao brick tea than in Pu'er brick tea (mean levels 441 and 77 mg/kg, respectively). According to these figures, the fluorine intake associated with consuming an infusion of 30 g Pu'er brick tea/person/day is safe because it does not exceed the maximum recommended daily allowance (RDA) of up to 4.0 mg for adults. In contrast, the almost six times higher intake of fluorine from Bianxiao brick tea greatly exceeds the 4 mg RDA and is unsafe. The difference in the fluorine levels of the two types of brick tea can be attributed to differences in the materials used to make them: Pu'er brick tea is made from tender leaves whereas Bianxiao brick tea is made from old tough leaves in which fluorine has accumulated. We conclude that consumption of Pu'er brick tea is unlikely to induce fluorosis, which has been associated with consumption of Bianxiao brick tea. PMID- 9862648 TI - Non-carcinogenicity of capsaicinoids in B6C3F1 mice. AB - The carcinogenicity of a mixture of capsaicinoids (64.5% capsaicin and 32.6% dihydrocapsaicin) was examined in B6C3F1 mice. In a 13-week toxicity study, renal toxicity was observed in 1% capsaicinoid-treated males. Next, groups of 50 mice of each sex were given 0, 0.025, 0.083 or 0.25% capsaicinoids in powdered diet for 79 weeks and killed in week 83. Food intake was reduced in mice of all capsaicinoid-treated groups, especially females, because of the pungency of capsaicinoids, and inhibition of body weight gain was apparent in females. The numbers of tumour-bearing females in the high-dose groups were significantly lower than that in the controls, and the incidences of hepatocellular neoplasms in both sexes were negatively correlated with the dose of capsaicinoids (Cochran Armitage trend test). Renal cell adenomas developed in one mouse each of 0.025 and 0.25% capsaicinoid-treated males. The incidences of other tumours were similar in the treated and control groups. Thus, the present study indicated that a mixture of capsaicinoids is not carcinogenic in B6C3F1 mice. PMID- 9862649 TI - Safety evaluation of Nostoc flagelliforme (nostocales, Cyanophyceae) as a potential food. AB - The safety of the alga genus Nostoc flagelliforme Born. etFlah. as a human food source was evaluated in an oral acute toxicity study and in a 28-day oral subacute toxicity study using rats. In the acute toxicity study, the dried powder of N. flagelliforme was orally administered to male and female rats at a dose of 1250 mg/kg and 2500 mg/kg. Neither mortality nor changes in general condition were observed in either the study groups or the control group over a 14-day observation period. In the subacute toxicity study, N. flagelliforme powder was administered orally to male and female rats at a dose of 500 mg/kg and 1000 mg/kg for a period of 28 days. Neither mortality nor changes in general condition were observed in either the treatment group or the control group throughout the 28-day administration period. No reduction in food consumption or body weight gain was observed in the experimental animals. Ophthalmological tests performed at the end of the administration period revealed no abnormalities within the ophthalmological parameters. In the haematological tests and serum biochemical tests performed at the time of completion of the administration period, no adverse effects of N. flagelliforme were observed. At autopsy, organ weight at the end of the experimental period and histopathological tests of specimens obtained from the autopsied animals revealed no significant influences of N. flagelliforme. In conclusion, considering the absence of adverse effects of N. flagelliforme in this study, findings in the oral acute toxicity study and the 28 day oral subacute toxicity study may indicate the safety of N. flagelliforme for human consumption. This study is in agreement with the novel nutraceutical idea "Phycophagism". PMID- 9862650 TI - Shark cartilage-containing preparation: protection against reactive oxygen species. AB - There is overwhelming evidence to indicate that free radicals cause oxidative damage to lipids, proteins and nucleic acids and are involved in the pathogenesis of several degenerative diseases. Therefore, antioxidants, which can neutralize free radicals, may be of central importance in the prevention of these disease states. The protection that fruits and vegetables provide against disease has been attributed to the various antioxidants contained in them. Recently, an anti inflammatory and analgesic activity of a water-soluble fraction from shark cartilage has been described. Using electrophoretical assays, bacteria survival and transformation and the Salmonella/mammalian-microsome assay, we investigated the putative role of shark cartilage-containing preparation in protecting cells against reactive oxygen species induced DNA damage and mutagenesis. If antimutagens are to have any impact on human disease, it is essential that they are specifically directed against the most common mutagens in daily life. Our data suggest that shark cartilage-containing preparation can play a scavenger role for reactive oxygen species and protects cells against inactivation and mutagenesis. PMID- 9862651 TI - Safety and tolerance of Lactobacillus reuteri supplementation to a population infected with the human immunodeficiency virus. AB - Probiotic supplementation may provide health benefits, especially for individuals with an underlying disease state that makes them more susceptible to infections. The purpose of this experiment was to evaluate the safety and tolerance of Lactobacillus reuteri ingestion by subjects infected with the human immunodeficiency virus (HIV). Thirty-nine subjects consumed a freeze-dried preparation of L. reuteri or a placebo for 21 days in a double-masked, parallel design experiment. Serum chemistry, haematology, immune profile, urinalysis, physical examination, gastrointestinal tolerance and faecal microbiota data were collected. No clinically significant changes were noted in any of the safety parameters measured. Overall, tolerance was good in both groups. Consumption of L. reuteri tended to increase faecal levels of L. reuteri on days 7, 14 and 21 of treatment feeding (P < 0.06, P < 0.11 and P = 0.05, respectively). However, faecal levels of L. reuteri and total Lactobacillus species were lower than levels previously observed in healthy male adults. Overall, this study documents that L. reuteri may be fed to HIV-positive individuals at 1 x 10(10) colony forming units/day without any clinically significant safety or tolerance problems. PMID- 9862652 TI - Experimental murine IgA nephropathy following passive administration of vomitoxin induced IgA monoclonal antibodies. AB - Oral exposure of mice to vomitoxin (VT) induces elevated levels of serum IgA, circulating IgA immune complexes (IgA-IC), mesangial IgA deposition and haematuria, which all mimic the clinical signs of human IgA nephropathy (IgAN). To further assess the effects of VT-induced IgA in the murine model, B6C3F1 and BALB/C mice were injected intraperitoneally with affinity-purified monoclonal IgA derived from Peyer's patch hybridomas of VT-exposed mice. In B6C3F1 mice, serum IgA, IgM and IgA-IC levels were increased two- to fivefold in treatment groups after 4 and 6 wk compared with controls, whereas increases in serum IgG as high as 18-fold were observed. Urinary erythrocyte counts were also significantly elevated in treatment groups after 2, 4 and 6 wk compared with controls. Concurrent increases in IgA and IgG complexes containing casein, the dietary protein source, occurred in treatment mice. Mesangial IgA, IgG, IgM and C3 deposition were significantly increased in all treatment mice after 6 wk. Electron-dense deposits occurred in the glomeruli of IgA-injected mice after 6 wk. All the above parameters were similarly affected in BALB/C mice. Injection of IgA-secreting hybridoma cells into BALB/C mice increased serum IgA, IgA-IC and IgG levels as well as elevated mesangial IgA, IgG and C3 deposition and haematuria after 2-3 weeks compared with controls. In total, these data indicate that passive administration of VT-induced IgAs can induce the hallmarks of IgA nephropathy. Casein, an antigen found in the diet used for these mice, appeared to form IC with IgA or IgG and these IC may participate in the pathogenesis of this nephropathy. PMID- 9862654 TI - Testing the potential of sodium fluoride to affect spermatogenesis: a morphometric study. AB - This study provides quantitative information on the effect of sodium fluoride (NaF) on the testes of F1 generation male rats exposed in utero and during lactation to NaF at one of four concentrations (25, 100, 175, 250 ppm). At weaning, the F1 generation males were exposed to NaF in their drinking water for 14 weeks, after which time testicular tissues were perfusion-fixed with glutaraldehyde and observed after being embedded in plastic. The seminiferous tubules comprised 89%, 87%, 88%, 88% and 88% of the total testis volume while the interstitial space occupied 9.3%, 11.2%, 10.2%, 9.8% and 9.9% of the total testis volume for the 0, 25, 100, 175 and 250 ppm NaF treatment groups, respectively. Statistically significant differences between control and NaF-treated rats were not observed with respect to absolute volume of the seminiferous tubules, interstitial space, Leydig cells, blood vessels boundary layer, lymphatic space, macrophages, tubular lumen or absolute tubular length and absolute tubular surface area, mean Sertoli cell nucleoli number per tubular cross-section, mean seminiferous tubule diameter and the mean height of the seminiferous epithelium. A statistically significant decrease in the absolute volume and volume percent of the lymphatic endothelium was observed in the 175 and 250 ppm NaF-treated groups and in the testicular capsule in the 100 ppm NaF-treated groups. The significance of this finding is unknown at the present time. Overall, the quantitative information obtained suggests that exposure to NaF at the doses used in the present study does not adversely affect testis structure or spermatogenesis in the rat. PMID- 9862653 TI - Immunological evaluation of the mycotoxin patulin in female B6C3F1 mice. AB - Patulin is a mycotoxin produced by many fungal species of the genera Penicillium, Aspergillus and Bryssochamys. Previous literature reports have suggested that patulin is toxic to the immune system. The studies presented were conducted to provide a comprehensive assessment of the effects of patulin on the immune system. Unlike previous reports, the doses of patulin used (0.08, 0.16, 0.32, 0.64, 1.28 and 2.56 mg/kg) were based on predicted human exposure levels. Female B6C3F1 mice were exposed orally to patulin for 28 days. Effects were not observed on final body weight or body weight gain. Relative weight of the liver, spleen, thymus, kidneys with adrenals, and lungs was not affected. Peripheral blood leucocyte and lymphocyte counts were decreased by approximately 30% in the two highest dose groups. The leucocyte differential was not altered. Total spleen cell, total T-cell (CD3+), helper T-cell (CD4+CD8-), B-cell (surface immunoglobulin+) and monocyte (MAC-3+) counts were not changed. Cytotoxic T-cell (CD8+CD4-) counts were increased 50% only by the highest dose. Natural killer cell (NK1.1+CD3-) and monocyte (MAC-1+) counts were increased 30% and 24%, respectively, only in the 0.08 mg/kg group. Humoral immune function as assessed by antibody-forming cell response and serum IgM titre to sheep erythrocytes, and cell-mediated immune function evaluated utilizing natural killer cell activity and the mixed lymphocyte reaction were not altered. Oral exposure to patulin for 28 days did not alter the ability of female B6C3F1 mice to mount either a cell mediated or humoral immune response. PMID- 9862655 TI - Carcinogenicity and co-carcinogenicity studies on propoxur in mouse skin. AB - Propoxur (2-isopropoxyphenyl methylcarbamate) is a widely used broad spectrum carbamate insecticide mainly used to control household pests. Propoxur exposure is reported to inhibit cholinesterase activity in rodents. Apart from other toxic effects, propoxur was found to possess tumorigenic activity in rats after oral administration. Propoxur does not produce tumours in mice or hamsters, or bladder hyperplasia in dogs and monkeys following oral feeding. In this set of investigations the complete carcinogenic, tumour initiating and promoting potential of propoxur was evaluated in male and female Swiss albino mice, since no information was available following dermal exposure of propoxur. The animals were exposed to propoxur through topical painting on the interscapular region at a dose of 100 mg/kg body weight. The results revealed that propoxur has tumour promoting potential on mouse skin following a two-stage initiation-promotion protocol, but it failed to induce the tumour(s) at a significant level, when tested for tumour initiating and complete carcinogenic property. PMID- 9862656 TI - Distribution of major and minor alkaloids in tobacco, mainstream and sidestream smoke of popular Indian smoking products. AB - Various Indian smoking products--cigarette, bidi, chutta and a brand of US cigarette--were analysed by gas chromatography-flame ionization detection (GC FID) for the levels of nicotine and minor tobacco alkaloids in tobacco, mainstream smoke (MS) and sidestream smoke (SS) employing modified smoking standards, namely two puffs/min. The analysis clearly demonstrated relatively higher levels of nicotine and minor tobacco alkaloids in tobacco from bidi (37.7 mg/g) and chutta (34.5 mg/g) when compared with Indian and US cigarettes (14-16 mg/g) studied. Relatively lower levels (SS/MS) of nicotine in SS from bidi and chutta compared with Indian/US cigarettes, suggest that the contribution of nicotine in SS from a single bidi/chutta to environmental tobacco smoke (ETS) is very much less than that of a single Indian/US cigarette. Reduced levels of nicotine in SS of bidi/chutta result in relatively higher deliveries of nicotine in MS as reflected by higher MS/SS values. The observed differences are likely to be due to difference in tobacco processing, burning rate/temperature and design of the smoking product. PMID- 9862657 TI - Erythritol: an interpretive summary of biochemical, metabolic, toxicological and clinical data. AB - A critical and comprehensive review of the safety information on erythritol was undertaken. Numerous toxicity and metabolic studies have been conducted on erythritol in rats, mice and dogs. The toxicity studies consist of long-term feeding studies conducted to determine carcinogenic potential, intravenous and oral teratogenicity studies to determine the potential for effects on the foetus, oral studies in which erythritol was administered over one or two generations to determine the potential for reproductive effects, and studies in bacterial and mammalian systems to determine mutagenic potential. The majority of the safety studies conducted were feeding studies in which erythritol was mixed into the diet at concentrations as high as 20%. The metabolic studies in animals have shown that erythritol is almost completely absorbed, not metabolized systemically and is excreted unchanged in the urine. The safety studies have demonstrated that erythritol is well tolerated and elicits no toxicological effects. The clinical program for erythritol involved a series of single-dose and repeat-dose, short duration studies which have been used to investigate the human correlates to the physiological responses seen in the preclinical studies. The clinical studies showed erythritol to be well tolerated and not to cause any toxicologically relevant effects, even following high-dose exposure. Erythritol administered orally to humans was rapidly absorbed from the gastrointestinal tract and quantitatively excreted in the urine without undergoing metabolic change. At high oral doses, urinary excretion accounted for approximately 90% of the administered dose with minimal amounts appearing in the faeces. A comparison of the human and animal data indicated a high degree of similarity in the metabolism of erythritol and this finding supports the use of the animal species used to evaluate the safety of erythritol for human consumption. It can be concluded, based on the available studies that erythritol did not produce evidence of toxicity. PMID- 9862658 TI - A current problem in the measurement of surgical wound infection rates. PMID- 9862659 TI - The time of presentation of wound infection after cardiac surgery. AB - Clinical experience indicates that many wound infections present relatively late after cardiac surgery. Hence, timing may be an important issue in using this outcome as a clinical indicator. A database of 1000 patients who underwent cardiac surgery was accessed to ascertain baseline characteristics, the type of surgery, and the time of presentation of wound infections. The overall incidence of wound infection was 5.9% (59/1000). Only 36% (21/59) of the wound infections presented while the patient was in hospital. Diabetics were more likely to have a late presentation of a wound infection (the median time of presentation of wound infections (more than 17 days), i.e. 10/29 (33.4%) versus 98/971 (10.1%), Fisher's exact test P < 0.01). Wound infection can only be regarded as a reliable clinical indicator after cardiac surgery if patients are reviewed with care for 6 weeks after surgery. PMID- 9862660 TI - Uncovering the causes of poor quality pathology laboratory services in acute myocardial infarction. AB - A study was undertaken to identify the causes that contributed to poor quality pathology services for hospitalized patients with acute myocardial infarction. Through the use of continuous quality improvement (CQI) strategies, the procedure involved a systematic process analysis which assessed the types and sources of variation, and possible causal factors for changes in performance. Thirteen suspected causes of poor quality were identified and data collected to confirm or reject their involvement. The study demonstrated that even strongly suspected causes of poor quality need to be confirmed by data. We found that 23% of the suspected root causes were not substantiated by the data collected. The CQI approach was process driven and should be adaptable to other clinical situations and not just pathology services. PMID- 9862661 TI - The quality of communication between hospitals and general practitioners: an assessment. AB - The objective of this study was to assess the quality of communications between hospitals and general practitioners (GPs). The proportion of medical records in which the patient's general practitioner (GP) was identified, the accuracy of medications recorded in the discharge summary, the proportion of GPs who received discharge summaries, and the timeliness of receipt of discharge summaries were all evaluated. Discussions were held with all stakeholders, the literature was reviewed and GPs were surveyed to identify potential measures of quality. These were then trialled to assess their utility and practicability. Timeliness, issues that required follow-up and treatment provided in hospital were of greatest importance to general practitioners. The GP's name was recorded in 88% of audited records. Few inaccuracies were detected in the medications recorded in the discharge summaries, and GPs received 77% of discharge summaries. Methods similar to those used in this study might be broadly applied to improve the quality of discharge communication throughout Australia. PMID- 9862662 TI - Incident monitoring in psychiatry. AB - Critical Incident Monitoring (CIM) as an instrument of quality assurance (QA) has received increasing attention in recent years. The present study was developed to explore a potential role for CIM in QA for clinical psychiatry. A questionnaire was sent to psychiatrists and requested retrospective reporting of clinical incidents, and a pilot study of an inpatient-based incident reporting system was performed. All Fellows of the Royal Australian and New Zealand College of Psychiatry (RANZCP) were sent a questionnaire. Eight psychiatric inpatient services were invited to participate in the pilot study. The returns of the questionnaires were aggregated and analysed to reveal a relatively small number of separate incident types, with little difference between the 'adverse outcome' and 'near-miss' categories. Similar results were found with the pilot study. It was concluded that the development of a unified incident reporting system for use by psychiatric clinicians and psychiatric services may add usefully to existing quality improvement processes. PMID- 9862663 TI - The impact of new technology on cholecystectomy rates in New South Wales. AB - This study examined the impact of the introduction of laparoscopic cholecystectomy (LC) on the rate and pattern of cholecystectomy in New South Wales, Australia. An analysis of the hospital data for the period 1981-94/95 was performed, which encompassed patients of both public and private hospitals. Cholecystectomy rates in the years after the introduction of LC in mid-1990 were 24% higher on average than rates in the prior years and rose from an average 1.68 2.09 per 1000. In the 5-year period of the study after the introduction of LC, the average age of private hospital patients increased from 50.6 to 53.4 years, while that of public hospital patients declined slightly from 53.5 to 52.4 years. Laparoscopic cholecystectomy was introduced more rapidly in private than public hospitals. By June 1995, 96% of private compared with 71% of public hospitals had introduced LC, and cholecystectomy patients in private hospitals were more likely to have laparoscopic surgery than patients in public hospitals (92% compared with 80%). The introduction of LC has been accompanied by a marked and sustained increase in the cholecystectomy rate. While the unit cost for the laparoscopic procedure is lower because of a shorter length of stay, the increase in the number of procedures means that there have been no net savings for the health system from the increased efficiency. While the new procedure offers superior advantages to patients with symptomatic gallbladder disease, the effect of the apparent lower threshold for surgery needs investigation. PMID- 9862664 TI - Suspected infection in children with cancer. AB - The microbiology and severity of suspected infections in children with cancer admitted to the John Hunter Children's Hospital was determined in order to assess whether any alteration to the infection treatment protocol was required. All children with cancer aged 1-17 years who had an episode of suspected or proven infection that required parenteral treatment at John Hunter Children's Hospital (JHCH) during 1994/95 were reviewed. Thirty-seven children were treated for cancer at JHCH; 62 admissions for suspected infection which involved 26 children were reviewed. Sixteen of these children had a permanent central line. Children with a central line had an increase in the number of days of inpatient treatment required for the treatment of suspected infection, and they had more episodes of infection. A pathogen was isolated more frequently with blood cultures being positive more often and gram-positive species were methicillin resistant more often. These differences were not statistically significant. A pathogen was isolated in 52% of admissions. Sixteen pathogens were gram positive; 12 were gram negative, two were fungal and two were viral. Blood cultures were positive in 21 of 62 admissions, skin swabs in four admissions, urine cultures in three admissions, stool in two admissions and one species was isolated from an epidural catheter tip and from the sputum. In 16% of admissions, the identified organism was resistant to the initial empirical therapy of tobramycin and piperacillin. In a further 13%, flucloxacillin was added to the empirical regimen when a sensitive Staphylococcus was identified. No significant differences between the culture negative and culture-positive groups were observed in admission pulse, fever or admission neutrophil count. However, those patients with a central line had a higher incidence of having a pathogen isolated if their temperature was > 39.5 degrees C. The median length of stay was longer for patients with a pathogen isolated on blood culture. Admission blood cultures were positive in 53% of admissions with an initial neutrophil count > 1000 x 10(9)/mL. Each of these children had a central line. Only one child died of infection during the 2-year study period. This review supports the observations that gram-positive infection is now more common than gram-negative infection in children with cancer. Despite the management advantages a permanent central line affords it is clear those children with a central line have an increased rate of infection and there needs to be caution in their use. The most important is the observation that any fever > 39.5 degrees C in a child with a central line is likely to be associated with a documented infection irrespective of the neutrophil count. The clinical outcomes observed in the present study indicate that tobramycin and piperacillin are effective empirical treatments for suspected infection in children with cancer. PMID- 9862665 TI - Virus-activated CD8 T cells and lymphokine-activated NK cells express the mast cell function-associated antigen, an inhibitory C-type lectin. AB - The mast cell function-associated Ag (MAFA) is an inhibitory C-type lectin that was originally identified on the cell surface of a rat mucosal mast cell line, RBL-2H3. We have cloned the mouse homologue of the rat MAFA gene, and Northern blot analysis revealed that mouse MAFA (mMAFA) gene expression was strongly induced in effector CD8 T cells and lymphokine-activated NK cells but not in effector CD4 T cells and in mouse mast cells. Moreover, mMAFA gene expression was only found in effector CD8 T cells that had been primed in vivo with live virus because in vitro activated CD8 T cells did not express mMAFA. Primary sequence comparison revealed a high degree of conservation (89% similarity) between rat MAFA and mMAFA. Thus, the MAFA molecule in the mouse is a putative inhibitory receptor on anti-viral CD8 T cells induced in vivo and on NK cells. PMID- 9862666 TI - Nuclear factor of activated T cells and AP-1 are insufficient for IL-2 promoter activation: requirement for CD28 up-regulation of RE/AP. AB - IL-2 gene transcription in T cells requires both TCR and costimulatory signals. IL-2 promoter activation in Jurkat T cells stimulated with superantigen presented by Raji B cells requires CD28 activation. The addition of rCTLA4Ig, which blocks CD28 binding to its ligand, to the cultures decreased IL-2 promoter activation by >80%. Interestingly, CTLA4Ig did not significantly inhibit the activation of either NF of activated T cells (NFAT) or AP-1 reporters. Therefore, activation of NFAT and AP-1 is insufficient for IL-2 promoter activation. In contrast, an RE/AP reporter was blocked by CTLA4Ig by >90%. Thus, the requirement for CD28 in IL-2 promoter activation appears to be due to RE/AP and not the NFAT or AP-1 sites. In addition, these data suggest that transcriptional activation of RE/AP is not mediated by NFAT, because activation of a NFAT reporter is not affected by the addition of CTLA4Ig. PMID- 9862667 TI - TCR signaling induces selective exclusion of CD43 from the T cell-antigen presenting cell contact site. AB - CD43, a large highly glycosylated molecule, is arguably the most abundant molecule on the surface of T cells. Nevertheless, the function of CD43 remains unclear. Utilizing fluorescence microscopy, we find that CD43 is excluded from the T cell-APC contact site. This exclusion is Ag dependent since optimal CD43 exclusion requires Ag-pulsed APC, and since signaling through CD3, in the absence of any other receptor ligand interactions, can induce the modulation of CD43. These data suggest that CD43 may function as a barrier to nonspecific T cell-APC interactions that is removed as a result of T cell activation. Exclusion from the interaction site is a unique feature of CD43 and not universally found for all large highly glycosylated molecules since CD45 is not excluded. Thus, CD43 may represent a novel regulatory molecule on the T cell surface that can direct T cell interactions by changing its location on the cell surface. PMID- 9862668 TI - Impaired mast cell development and innate immunity in Mac-1 (CD11b/CD18, CR3) deficient mice. AB - Mac-1 (CD11b/CD18, CR3), a beta2 integrin expressed on leukocytes, is important in leukocyte migration. We demonstrate that Mac-1 is also expressed on peritoneal mast cells and LPS stimulated bone marrow-derived cultured mast cells, and that Mac-1-deficient mice, which lack this receptor, have significant reductions in the numbers of mast cells resident in the peritoneal cavity, peritoneal wall, and dorsal skin. The reduced numbers of mast cells in Mac-1-deficient mice may have important functional consequences, in that Mac-1-deficient mice exhibit significantly increased mortality after cecal ligation and puncture, a model of acute septic peritonitis in which host resistance has been shown to be dependent on both mast cells and complement. These findings demonstrate that Mac-1 is required for the expression of normal levels of mast cells in the peritoneal cavity, peritoneal wall, and certain areas of the skin, as well as for maintaining adequate mast cell-dependent host defense against bacterial infection. PMID- 9862669 TI - Increased activity of oleate-dependent type phospholipase D during actinomycin D induced apoptosis in Jurkat T cells. AB - Apoptosis is an active form of cell death that can be induced by a wide variety of agents and conditions. In response to actinomycin D, hydrogen peroxide (H2O2), or TNF-alpha, Jurkat T cells underwent typical apoptosis. Phospholipase D (PLD) activity in intact cells determined by phosphatidylbutanol generation was up regulated by these agents. The PLD activation was in a time-dependent manner during apoptosis. It was also shown that the PLD activity measured by using exogenous substrate in the lysate from apoptotic cells was higher than that in the lysate from control untreated cells. The PLD activity in lysate from control untreated cells was stimulated by unsaturated fatty acids (UFA), but not by guanosine 5'-O-(3-thiotriphosphate). However, the PLD activity in the apoptotic cell lysate was no longer enhanced by the addition of oleate, suggesting that the increased PLD activity during apoptosis was attributed to the PLD of UFA dependent type, but not the small G protein-dependent one. In fact, the release of free UFA was increased during apoptosis. The caspase inhibitors, z-DEVD and z VAD, effectively suppressed PLD activation and apoptosis, but UFA release was unaffected. These results suggest the possibility that UFA-dependent type PLD may be implicated in apoptotic process in Jurkat T cells. This is the first demonstration that the PLD of UFA-dependent type would be involved in cellular responses. PMID- 9862670 TI - MHC class I mosaic mice reveal insights into control of Ly49C inhibitory receptor expression in NK cells. AB - We have analyzed lymphocyte development in natural MHC class I chimeric mice, generated through a transgenic approach in beta2-microglobulin (beta2m)-/- mice. In these mice, MHC class I+ cells coexist with an equal proportion of MHC class I deficient cells. These MHC class I mosaic mice had normal numbers of CD8+ T cells, which had a target cell specificity similar to that of wild-type mice. Consequently, the mice did not develop any signs of autoimmunity. They also had normal numbers of NK cells. This allowed an examination of the MHC class I influence on the expression of the Ly49C inhibitory receptor on NK cells. This receptor binds to H-2Kb. It is expressed at low levels on NK cells in wild-type mice of the H-2b haplotype, but at markedly higher levels on NK cells in beta2m-/ mice and other strains of mice lacking expression of H-2Kb. Relatively little is known about how MHC class I molecules affect expression of the Ly49 receptors. Through the analysis of the present MHC class I mosaic mice, we demonstrate that the expression levels of Ly49C on NK cells is a consequence not only of MHC class I expression in the environment, but also of the expression of MHC class I molecules by the NK cells themselves. These findings are discussed in relation to the biological role of the calibration of the Ly49 inhibitory receptor expression in relation to self-MHC class I. PMID- 9862671 TI - IL-6-deficient mice are resistant to experimental autoimmune encephalomyelitis: roles of IL-6 in the activation and differentiation of autoreactive T cells. AB - Although autoreactive T cells recognizing self myelin Ags are present in most individuals, autoimmune disease of the central nervous system is a relatively rare medical condition. Development of autoimmune disease may require not only the presence of autoreactive T cells but also that autoreactive T cells become activated. Activation of T cells may require a minimum of two signals: an Ag specific signal delivered by MHC-peptide complex and a second signal delivered by costimulatory molecules or cytokines. Although in vitro studies have suggested that cytokines, especially proinflammatory cytokines such as IL-1, IL-6, and TNF are involved in T cell activation, their precise roles in vivo are not clear. To determine the roles of proinflammatory cytokines in T cell activation in vivo and in the development of autoimmune disease, we have studied experimental autoimmune encephalomyelitis (EAE) in mice deficient in IL-6. We found that IL-6-deficient mice were completely resistant to EAE induced by myelin oligodendrocyte glycoprotein (MOG), whereas IL-6-competent control mice developed EAE characterized by focal inflammation and demyelination in the central nervous system and deficiency in neurologic functions. Furthermore, we established that the resistance to EAE in IL-6-deficient mice was associated with a deficiency of MOG-specific T cells to differentiate into either Th1 or Th2 type effector cells in vivo. These results strongly suggest that IL-6 plays a crucial role in the activation and differentiation of autoreactive T cells in vivo and that blocking IL-6 function can be an effective means to prevent EAE. PMID- 9862672 TI - Inside the crawling T cell: leukocyte function-associated antigen-1 cross-linking is associated with microtubule-directed translocation of protein kinase C isoenzymes beta(I) and delta. AB - T cells activated via integrin receptors can polarize and start crawling locomotion with repeated cycles of cytoskeletal reassembly processes, many of which depend on phosphorylation. We demonstrate that protein kinase C (PKC) activation represents an essential event in induction of active T cell motility. We find that in crawling T cells triggered via cross-linking of integrin LFA-1 two PKC isoenzymes, beta(I) and delta, are targeted to the cytoskeleton with specific localization corresponding to the microtubule-organizing center (MTOC) and microtubules, as detected by immunocytochemistry and immunoblotting. Clustering of LFA-1 associated with its signaling function also occurs at the membrane sites adjacent to the MTOC. We further show that cells of a PKC-beta deficient clone derived from parental PKC-beta-expressing T cell line can neither crawl nor develop a polarized microtubule array upon integrin cross-linking. However, their adhesion and formation of actin-based pseudopodia remain unaffected. Our data demonstrate the critical importance of the microtubule cytoskeleton in T cell locomotion and suggest a novel microtubule-directed intracellular signaling pathway mediated by integrins and involving two distinctive PKC isoforms. PMID- 9862674 TI - Stimulation of Stat5 by granulocyte colony-stimulating factor (G-CSF) is modulated by two distinct cytoplasmic regions of the G-CSF receptor. AB - In a manner similar to many other cytokines, treatment of cells with granulocyte CSF (G-CSF) has been shown to induce the tyrosine phosphorylation of the STAT proteins. Activation of Stat1 and Stat5 by G-CSF requires the membrane-proximal cytoplasmic domain of the receptor, including box1 and box2, while G-CSF stimulated tyrosine phosphorylation of Stat3 also requires a region distal to box 2. In this study, we show that although the membrane-proximal 55 amino acids of the G-CSF receptor are sufficient for activation of Stat5, the maximal rate of Stat5 activation requires an additional 30 amino acids of the cytoplasmic domain. In contrast, the distal carboxyl-terminal region of the receptor appears to down regulate Stat5 activation in that deletion of this carboxyl terminus results in increased amplitude and prolonged duration of Stat5 activation by G-CSF. Significantly, expression of a truncated dominant-negative Stat5 protein in hemopoietic cells not only inhibits G-CSF-dependent cell proliferation, but also suppresses cell survival upon G-CSF withdrawal. We further show that a potential protein tyrosine phosphatase may play a critical role in the down-regulation of G CSF-stimulated Stat5 activation. These results demonstrate that two distinct cytoplasmic regions of the G-CSF receptor are involved in the regulation of the intensity and duration of Stat5 activation, and that Stat5 may be an important player in G-CSF-mediated cell proliferation and survival. PMID- 9862673 TI - CD27/CD70 interaction augments IgE secretion by promoting the differentiation of memory B cells into plasma cells. AB - The induction of IgE switching in B cells requires several signals given by cytokines and cell contact-delivered signals. Here, we investigated the role of CD27/CD70 interaction in B cell IgE synthesis. The addition of CD27 ligand (CD70) transfectants to B cell cultures increased the IgE synthesis synergistically in the presence of IL-4 plus anti-CD40 mAb (anti-CD40). The effect of CD70 transfectants was dose dependent and was completely blocked by anti-CD70 mAb. CD27+ B cells had the ability to produce IgE, which was increased by contact with CD70 transfectants, whereas CD27- B cells did not produce IgE. CD27/CD70 interaction enhanced B cell proliferation in the presence of IL-4 or IL-4 plus anti-CD40. The augmentation of B cell proliferation by CD70 transfectants was apparent in CD27+ B cells, but was mild in CD27- B cells. The helper activity for IgE synthesis by the CD27/CD70 interaction did not contribute to the enhancement of germline epsilon transcripts. Flow cytometric and morphological analyses demonstrated that the addition of CD70 transfectants to B cell cultures remarkably promoted differentiation into plasma cells in the presence of IL-4 and CD40 signaling. Finally, CD27 cross-linking resulted in the up-regulation of positive regulatory domain I-binding factor-1. Taken together, our findings indicate that signaling via CD27 on B cells induces IgE synthesis, in cooperation with IL-4 and CD40 signaling, by promoting the generation of plasma cells through up-regulation of positive regulatory domain I-binding factor-1. PMID- 9862675 TI - Ox-40 ligand: a potent costimulatory molecule for sustaining primary CD4 T cell responses. AB - Ox-40 and Ox-40 ligand (Ox-40L) are thought to be involved in T cell-APC interactions. However, their exact role in T cell responses is undefined. Using fibroblast transfectants expressing Ox-40L and/or B7-1, and CD4 cells from TCR transgenic mice, we investigated the effect of Ox-40 signaling on primary responses to the Ag pigeon cytochrome c. Ox-40 expression on naive CD4 cells peaked 2 to 3 days after activation, and was lost by 4 to 5 days. APCs with Ox 40L promoted partial activation of naive T cells with some IL-2 secretion, but were unable to enhance proliferation, unlike those with B7-1. APCs coexpressing Ox-40L with B7-1 induced large quantities of IL-2 and promoted proliferative responses that persisted for several days. Effector cells taken 5 days after naive T cell activation reexpressed Ox-40 within 4 h and responded strongly to APCs expressing Ox-40L, whereas B7-1 had little effect. Synergy was also seen between Ox-40L and B7-1, with primarily IL-2 being elevated, although IL-4 and IL 5 were also up-regulated. The most striking action was on effector T cell proliferation, which continued at high levels for up to 4 days, with little proliferation evident at this time in the absence of Ox-40 signals. These data suggest that Ox-40/Ox-40L interactions act after initial activation events to prolong clonal expansion and enhance effector cytokine secretion, and may be involved in promoting long-lived primary CD4 responses. PMID- 9862676 TI - Epitope-specific antibody and suppression of autoantibody responses against a hybrid self protein. AB - This study addresses the relationship of epitope-specific Ab responses and alternative autoantibody responses in a model system in which an antigenized self protein serves as the carrier for a defined heterologous B cell epitope. Ubiquitin, a nonimmunogenic self protein, was engineered to present heterologous B and T cell epitopes in the recombinant molecule. Fusion to the C terminus introduced a universal T cell epitope from a Mycobacterium tuberculosis Ag. The B cell epitope was created by inserting a 12-residue loop sequence of HIV-1 gp120 at a surface-exposed position of ubiquitin. These modifications preserved the ubiquitin fold, allowing a new conformational epitope to be presented among native self epitopes. Mice immunized with the hybrid protein bearing only the mycobacterial T cell epitope elicited a strong autoantibody response to native ubiquitin. In contrast, antisera elicited against hybrid ubiquitin presenting the HIV B cell epitope reacted specifically with the foreign epitope but not with native ubiquitin. Absence of autoantibody in the response was attributed to poor competition of autoreactive B cells for limiting T cell help. Both types of responses were associated with Th responses to defined epitopes of the ubiquitin hybrid protein. These results may have implications for a tolerance mechanism dependent on B-T cell cooperation. PMID- 9862677 TI - Nitric oxide synthase plays a signaling role in TCR-triggered apoptotic death. AB - A functional role for stimulated nitric oxide (NO) production was tested in the TCR-triggered death of mature T lymphocytes. In purified peripheral human T cell blasts or the 2B4 murine T cell hybridoma, apoptotic cell death induced by immobilized anti-CD3 was blocked by inhibitors of NO synthase (NOS) in a stereospecific and concentration-dependent manner. This effect appeared to be selective since apoptotic death induced by anti-Fas Ab or the steroid dexamethasone was not affected by NOS inhibitors. TCR-stimulated expression of functional Fas ligand was attenuated in a stereospecific manner by NOS inhibitors, but these compounds did not inhibit TCR-stimulated IL-2 secretion or CD69 surface expression. Nitrosylated tyrosines, a stable marker for NO generation, were immunochemically detected in T cells using flow cytometry. TCR signals induced NO production, as measured by an increase in nitrotyrosine specific staining. NOS enzymatic activity was detected in lysates of 2B4 cells, and Western blot analysis suggests that the activity is due to expression of the neuronal isoform of NOS. Thus, T cells have the capacity to generate NO upon Ag signaling, which may affect signal transduction, Fas ligand surface expression, and apoptotic cell death of mature T lymphocytes. PMID- 9862678 TI - Cytotoxic T cell responses to DNA vaccination: dependence on antigen presentation via class II MHC. AB - This study was designed to test whether cytotoxic T cell (CTL) responses to DNA vaccination are dependent upon MHC class II-restricted priming of CD4+ T cells. Because DNA vaccination may directly transfect dendritic cells, and dendritic cells may be capable of directly stimulating CD8+ T cell responses, such priming might be unnecessary. To test this hypothesis, C57BL/6 mice were immunized intramuscularly or intradermally with DNA encoding either whole OVA, a class I (Kb)-restricted peptide epitope of OVA (amino acids 257-264, SIINFEKL), or this class I-restricted epitope plus the adjacent class II (I-Ab)-restricted epitope of OVA (amino acids 265-280, TEWTSSNVMEERKIKV). Very low to negligible CTL responses were observed in mice vaccinated with the SIINFEKL construct, whereas mice vaccinated with the SIINFEKLTEWTSSNVMEERKIKV or with the complete OVA construct made equally robust CTL responses. These responses were sensitive to blocking by anti-CD8 mAb and were shown to be SIINFEKL-specific by using SIINFEKL peptide-pulsed EL-4 cells as targets. To ensure that the generation of these CTL responses was indeed dependent upon CD4+ T cell help, mice were depleted of either CD4+ or CD8+ cells before immunization. Depletion of CD4+ cells completely abrogated the CTL response to OVA DNA, as did depletion of CD8+ cells. Thus, we conclude that the CTL response to both intramuscular and intradermal DNA vaccination is highly dependent upon the generation of CD4+ T cell help via a class II MHC-dependent pathway. These results will be relevant for the construction of minimal-epitope vaccines for DNA immunization. PMID- 9862680 TI - Regulation of NK1.1 expression during lineage commitment of progenitor thymocytes. AB - We recently identified a stage in fetal ontogeny (NK1.1+/CD117+) that defines committed progenitors for T and NK lymphocytes. These cells are found in the fetal thymus as early as day 13 of gestation, but are absent in the fetal liver. Nonetheless, multipotent precursors derived from both the fetal thymus and fetal liver are capable of rapidly differentiating to the NK1.1+ stage upon transfer into fetal thymic organ culture (FTOC). This suggests that expression of NK1.1 marks a thymus-induced lineage commitment event. We now report that a subset of the most immature fetal thymocytes (NK1.1-/CD117+) is capable of up-regulating NK1.1 expression spontaneously upon short-term in vitro culture. Interestingly, fetal liver-derived CD117+ precursors remain NK1.1- upon similar culture. Spontaneous up-regulation of NK1.1 surface expression is minimally affected by transcriptional blockade, mitogen-induced activation, or exposure of these cells to exogenous cytokines or stromal cells. These data suggest that induction of NK1.1 expression on cultured thymocytes may be predetermined by exposure to the thymic microenvironment in vivo. Importantly, multipotent CD117+ thymocytes subdivided on the basis of NK1.1 expression after short-term in vitro culture show distinct precursor potential in lymphocyte lineage reconstitution assays. This demonstrates that even the earliest precursor thymocyte population, although phenotypically homogeneous, contains a functionally heterogeneous subset of lineage-committed progenitors. These findings characterize a thymus-induced pathway in the control of lymphocyte lineage commitment to the T and NK cell fates. PMID- 9862679 TI - Complement opsonization is required for presentation of immune complexes by resting peripheral blood B cells. AB - Complement receptor 2 (CD21, CR2) is a B cell receptor for complement degradation products bound to Ag or immune complexes. The role of CD21 in mediating Ag presentation of soluble immune complexes by resting B cells was studied. Complement-coated immune complexes were formed by the incubation of influenza virus with serum from immune donors. These complexes bound to peripheral blood B cells in a complement-dependent manner. The binding required CD21 or, to a lesser extent, complement receptor 1 (CR1, CD35). B cells pulsed with immune complexes containing complement elicited a response from a panel of influenza-specific T cell clones, while those pulsed with immune complexes formed in the absence of complement did not. The expression of the early activation marker CD69 and the costimulatory molecule CD86 were not induced by CD21 ligation alone, suggesting that CD21-mediated Ag presentation occurs independently of B cell activation. Up regulation of these markers required exposure to T cell factors elicited by the recognition of Ag derived from complement-containing immune complexes. These findings suggest that binding of Ag to CD21 enables Ag-nonspecific B cells to participate in the activation of Ag-specific T cells in a process that occurs independently of well-characterized B cell activation events. PMID- 9862681 TI - Importance of B7-1-expressing host antigen-presenting cells for the eradication of B7-2 transfected P815 tumor cells. AB - We have previously shown that B7-2 (CD86)-transfected P815 tumor cells elicit tumor-eradicating immunity that leads to the regression of the B7-2+ P815 tumor after transient growth in normal DBA/2 mice. Here, we show that both the B7-2 and B7-1 (CD80) molecules contribute to the eradication of B7-2+ P815 tumors as treatment of the mice with both anti-B7-2 and anti-B7-1 mAb was required to prevent B7-2+ P815 tumor regression. The cells that expressed the B7-1 molecule following inoculation of B7-2+ P815 tumor cells into normal mice were not the tumor cells but rather host APCs including MAC-1+ cells present in the draining lymph nodes. Moreover, B7-1-expressing host APCs were found to be important for the rejection of B7-2+ P815 tumors as anti-B7-2 mAb alone, which was ineffective in preventing B7-2+ P815 tumor rejection by normal wild-type mice, was effective in preventing B7-2+ P815 tumor rejection by mice in which the B7-1 gene was disrupted. Finally, consistent with the importance of B7-1-expressing host APCs for the generation of tumor-eradicating immunity against B7-2+ P815 tumor cells, CD4+ T cells (not only CD8+ T cells) were found to participate in tumor eradicating immunity against B7-2+ P815 tumor cells. Thus, in addition to eliciting tumor-eradicating immunity directly, B7-2+ P815 tumor cells elicit tumor-eradicating immunity indirectly through B7-1-expressing host APCs that present tumor-associated Ags to CD4+ T cells. PMID- 9862682 TI - Coreceptor-independent T cell activation in mice expressing MHC class II molecules mutated in the CD4 binding domain. AB - We have previously reported that efficient selection of the mature CD4+ T cell repertoire requires a functional interaction between the CD4 coreceptor on the developing thymocyte and the MHC class II molecule on the thymic epithelium. Mice expressing a class II protein carrying the EA137/VA142 double mutation in the CD4 binding domain develop fewer than one-third the number of CD4+ T cells found in wild-type mice. In this report we describe the functional characteristics of this population of CD4+ T cells. CD4+ T cells that develop under these conditions are predicted to be a CD4-independent subset of T cells, bearing TCRs of sufficient affinity for the class II ligand to undergo selection despite the absence of accessory class II-CD4 interactions. We show that CD4+ T cells from the class II mutant mice are indeed CD4 independent in their peripheral activation requirements. Surprisingly, we find that CD4+ T cells from the class II mutant mice, having been selected in the absence of a productive class II-CD4 interaction, fail to functionally engage CD4 even when subsequently provided with a wild-type class II ligand. Nevertheless, CD4+ T cells from EA137/VA142 class II mutant mice can respond to T-dependent Ags and support Ig isotype switching. PMID- 9862683 TI - The role of Stat4 in species-specific regulation of Th cell development by type I IFNs. AB - Type I IFNs (IFN-alpha/beta), in addition to IL-12, have been shown to play an important role in the differentiation of human, but not mouse, Th cells. We show here that IFN-alpha/beta act directly on human T cells to drive Th1 development, bypassing the need for IL-12-induced signaling, whereas IFN-alpha cannot substitute IL-12 for mouse Th1 development. The molecular basis for this species specificity is that IFN-alpha/beta activate Stat4 in differentiating human, but not mouse, Th cells. Unlike IL-12, which acts only on Th1 cells, IFN-alpha/beta can activate Stat4 not only in human Th1, but also in Th2 cells. However, restimulation of human Th2 lines and clones in the presence of IFN-alpha does not induce the production of IFN-gamma. These results suggest that activation of Stat4, which is necessary for the differentiation of naive T cells into polarized Th1 cells, is not sufficient to induce phenotype reversal of human Th2 cells. PMID- 9862684 TI - Antagonistic roles for phospholipase D activities in B cell signaling: while the antigen receptors transduce mitogenic signals via a novel phospholipase D activity, phosphatidylcholine-phospholipase D mediates antiproliferative signals. AB - Cross-linking of the Ag receptors on B cells induces DNA synthesis and proliferation. Butanol trap experiments suggest that one or more phospholipase D activities play a key role in this process. Although phosphatidylcholine phospholipase D has been shown to play a central role in the transduction of proliferative responses for a wide variety of calcium-mobilizing receptors, we show that the Ag receptors are not coupled to this phospholipase. In addition, phosphatidylcholine-phospholipase D is not stimulated under conditions that mimic T cell-dependent B cell activation. In contrast, ATP, which inhibits surface Ig (sIg)-mediated DNA synthesis in murine B cells via P2-purinoceptors, activates phosphatidylcholine-phospholipase D. Phosphatidylcholine-phospholipase D is therefore associated with antiproliferative signal transduction in mature B cells, but it does not transduce early signals associated with sIg-mediated growth arrest or apoptosis in immature B cells. Mitogenic stimulation of sIg is, however, coupled to a novel nonphosphatidylcholine-hydrolyzing phospholipase D activity. The resultant sIg-generated phosphatidic acid, unlike the phosphatidylcholine-derived phosphatidic acid generated via the purinoceptors, is converted to diacylglycerol. These data provide the first evidence that while the novel sIg-coupled phospholipase D and resultant diacylglycerol generation may play a role in B cell survival and proliferation, phosphatidylcholine phospholipase D may transduce, via phosphatidic acid, negative immunomodulatory signals in mature B lymphocytes. PMID- 9862685 TI - Induction or protection from experimental autoimmune encephalomyelitis depends on the cytokine secretion profile of TCR peptide-specific regulatory CD4 T cells. AB - Autoimmune diseases can result from the breakdown of regulation and subsequent activation of self-antigenic determinant-reactive T cells. During the evolution of the autoimmune response to myelin basic protein (MBP) in B10.PL mice, several distinct T cell populations expand: the effectors mediating experimental autoimmune encephalomyelitis (EAE) are MBP-reactive, CD4+, and predominantly TCR Vbeta8.2+; in addition, at least two regulatory populations can be detected--one comprised of Vbeta14+ CD4 T cells, reactive to a framework region 3 determinant on the Vbeta8.2 chain, and a second that is CD8+ and reactive to another Vbeta8.2 determinant. The combined action of these two regulatory cell types controls disease-causing effectors, resulting in spontaneous recovery from disease. In this report, we reveal that the cytokine secretion pattern of TCR peptide specific regulatory CD4 T cells can profoundly influence whether a type 1 or type 2 population predominates among MBP-specific CD4 effectors. The priming of type 1 regulatory T cells results in deviation of the Ag-specific effector T cell population in a type 2 direction and protection from disease. In contrast, induction of type 2 regulatory T cells results in exacerbation of EAE, poor recovery, and an increased frequency of type 1 effectors. Thus, the encephalitogenic potential of the MBP-reactive effector population is crucially and dominantly influenced by the cytokine secretion phenotype of regulatory CD4 T cells. These findings have important implications in understanding peripheral tolerance to self-Ags as well as in the design of TCR-based therapeutic approaches. PMID- 9862687 TI - Differential effects of peptide diversity and stromal cell type in positive and negative selection in the thymus. AB - Thymocyte positive selection results in maturation to the single-positive stage, while negative selection results in death by apoptosis. Although kinetic analyses indicate only 3-5% of CD4+ 8+ cells reach the single-positive stage, the balance of positive and negative selection and the nature and quantity of cells mediating maximal negative selection are uncertain. Here, using a system where the number and type of stromal cells and thymocytes can be controlled, we investigated the maturation of CD4+ 8+ thymocytes in the presence or absence of thymic epithelium and dendritic cells (DC) from wild-type (wt) and H-2M(-/-) mice expressing different peptide arrays. We find that titration of wt DC into reaggregates of wt epithelium has a dramatic effect on the number of CD4+ cells generated, with 1% DC causing a maximal 80% reduction. Moreover, while addition of 1% wt DC into cultures of H-2M(-/-) epithelium causes a 90% reduction in CD4+ cells, no effect was observed when similar numbers of wt thymic epithelium were added. Collectively, these data provide the first accurate indication of the quantity and quality of stromal cells required for maximal negative selection in the thymus, demonstrate the importance of peptide diversity in T cell selection, and highlight a large degree of overlap between positive and negative selection events. PMID- 9862686 TI - The development of autoimmune inflammatory arthropathy in mice transgenic for the human T cell leukemia virus type-1 env-pX region is not dependent on H-2 haplotypes and modified by the expression levels of Fas antigen. AB - Previously, we reported that human T cell leukemia virus type-1 env-pX region introduced transgenic (pX-Tg) mice develop an inflammatory polyarthropathy. Although autoimmune pathogenesis was suggested, the detailed mechanisms remain to be elucidated. In this report, we examined effects of the MHC and fas genes on the development of the disease. When pX-Tg mice were backcrossed with different inbred strains, the incidence of arthritis differed among strains; 64% and 72% in BALB/cAn (H-2d), 25% and 46% in C3H/HeN (H-2k), and 0% and 2% in C57BL/6J (H-2b) background at 3 and 6 months of age, respectively. Rheumatoid factor levels in the serum correlated with the susceptibility to the disease, whereas IL-1beta and MHC gene expression were similarly elevated in all of these strains, suggesting involvement of immune regulatory genes in this strain difference. However, introduction of the H-2d locus into C57BL/6J pX-Tg mice did not increase the incidence of arthritis, and substitution of the BALB/cAn H-2 locus with the H-2b did not decrease it. The results indicate that the H-2 locus is not the major determinant of the disease. Then, since previous study indicated a defect in Fas mediated apoptosis of transgenic T cells, the effects of fas gene modification on the disease were examined. The incidence increased when these pX-Tg mice were crossed with lpr/lpr mice, while it decreased when crossed with fas-transgenic mice. These observations suggest that aberration of Fas-mediated apoptosis of peripheral lymphocytes, rather than negative selection in the thymus, is involved in the development of autoimmune arthropathy in pX-Tg mice. PMID- 9862689 TI - Differential effects of CD28 engagement and IL-12 on T cell activation by altered peptide ligands. AB - To futher our understanding of the mechanisms underlying the diverse effects of altered peptide ligands (APL) on T cell activation, we used a population of nonactivated spleen cells from mice that expressed a transgenic TCR specific for myelin basic protein Ac1-11 and peptide analogues that display either enhanced or decreased affinities for TCR/MHC to address the question whether APL-induced signaling through the TCR can regulate the capability of APC to activate T cells. We demonstrate that weak agonists APL are poor inducers of all aspects of the activation of both the responder T cells and the APC. Enhancement of the antigenic signal by augmenting the binding of the weak agonists to MHC reversed their defective activating capacity. Enhancement of costimulation by engagement of CD28 only resulted in augmentation of the capacity of the weak agonist APL to induce proliferation and IL-2/IL-3 production, but not CD40L or IL-12Rbeta2 chain expression on T cells, CD80/CD86 expression on APC, IL-12 secretion, or IFN-gamma production. Exogenous IL-12 promoted IFN-gamma production in the presence of the weak agonists. These studies demonstrate that there is a critical threshold of antigenic signal required for full activation of the T cell-APC interactions needed for the differentiation of Th1 cells. The provision of excess costimulation can overcome some of the defects in T cell activation by weak agonists, but is insufficient to induce a sufficient level of CD40L expression needed for engagement of CD40 on APC with subsequent IL-12 production and induction of IL-12Rbeta2 chain expression. PMID- 9862688 TI - Loss of original antigenic specificity in T cell hybridomas transduced with a chimeric receptor containing single-chain Fv of an anti-collagen antibody and Fc epsilonRI-signaling gamma subunit. AB - T cell hybridomas HCQ6 and MD.45 acquired Ab-type specificity to collagen type II, when engrafted with a chimeric cell surface receptor, scC2Fv/gamma, which includes the single-chain Fv domain (scFv) of the anti-collagen type II mAb C2 and the signaling gamma subunit of the Fc epsilonRI. When transduced into MD.45 cells, scC2Fv/gamma or its mutated form lacking immunoreceptor tyrosine-based activation motif (ITAM), scC2Fv/gammaIC-, formed mainly homodimers. A small proportion of these molecules formed heterodimers with endogenous CD3zeta in these hybridoma cells. By contrast, in HCQ6 cells, the majority of scC2Fv/gamma and scC2Fv/gammaIC- molecules formed heterodimers with CD3zeta, and only a small proportion of them was expressed as homodimers. Stimulation with plastic immobilized collagen induced IL-2 production in scC2Fv/gamma-transduced MD.45 cells, but not in MD.45 cells transduced with the ITAM-less chimera scC2Fv/gammaIC-. HCQ6 cells transduced with scC2Fv/gamma responded to plastic bound collagen. Due to the high content of CD3zeta-associated chimeras, HCQ6 cells transduced with the ITAM-less scC2Fv/gammaIC- chimera were also responsive to plastic-bound collagen. When cells were stimulated with collagen in solution, MD.45 cells transduced with scC2Fv/gamma produced IL-2, whereas transduced HCQ6 cells were unresponsive, hence suggesting that the ability of cells transduced with scC2Fv chimeras to respond to soluble collagen correlated with predominant expression of divalent scC2Fv/gamma homodimers, but not monovalent scC2Fv/gamma CD3zeta or scC2Fv/gammaIC(-)-CD3zeta heterodimers. Of interest, expression of CD3 subunits in hybridomas transduced with scC2Fv chimeras was reduced, resulting in decreased response to cognate Ags. PMID- 9862690 TI - T cell proliferation-augmenting activities of the gene 3 protein derived from a phage library clone with CD80-binding activity. AB - We have isolated a phage clone, F2, by panning a phage library with a CTLA4 conformation recognizing mAb (anti-CTLA4 mAb). The unique sequence of 15 amino acids with an internal disulfide bond was inserted in the gene 3 proteins of F2 phage clone (F2-g3p). We show here that 1) F2-g3p was recognized with anti-CTLA4 mAb but not with anti-CD28 mAb, and 2) F2-g3p bound to CD80 but not to CD86. The surface plasmon resonance analysis showed that F2-g3p strongly bound CD80. F2-g3p inhibited the binding of CTLA4 to CD80 but not to CD86. In contrast, F2-g3p weakly inhibited the binding of CD28 with CD80. When hen egg lysozyme (HEL) primed lymph node cells were stimulated with HEL in the presence of F2-g3p in vitro, cell proliferation was highly potentiated. In the absence of antigenic stimulation, F2-g3p induced no T cell proliferation, indicating the costimulatory nature of F2-g3p. The T cell-augmenting activity of the F2 clone was eliminated when the F2 clone was preincubated with CD80-Ig before the addition to the cultures, indicating the involvement of CD80-binding in the F2-g3p-mediated immunopotentiation. Thus, the F2 motif conferred CD80-binding activity and an immunoregulatory function to the g3p. PMID- 9862691 TI - Selection and function of CD4+ T lymphocytes in transgenic mice expressing mutant MHC class II molecules deficient in their interaction with CD4. AB - Interactions of the T cell coreceptors, CD4 and CD8, with MHC molecules participate in regulating thymocyte development and T lymphocyte activation and differentiation to memory T cells. However, the exact roles of these interactions in normal T cell development and function remain unclear. CD4 interacts with class II MHC7 molecules via several noncontiguous regions in both the class II MHC alpha- and beta-chains. We have introduced a double mutation that disrupts interaction with CD4 into the I-A(beta)k gene and used this construct to generate transgenic mice expressing only mutant class II MHC. Although CD4+ thymocytes matured to the single-positive stage in these mice, their frequency was reduced by threefold compared with that of wild-type transgenics. Positive selection of CD4+ T cells in the mutant transgenic mice may have been mediated by TCRs with a higher than usual affinity for class II MHC/Ag complexes. In A(beta)k mutant transgenics, peripheral CD4+ lymphocytes promoted B cell differentiation to plasma cells. These CD4+ T cells also secreted IFN-gamma in response to various stimuli (e.g., protein Ag, bacterial superantigen, and alloantigen), but were deficient in IL-2 secretion. Interactions between CD4 and class II MHC molecules appeared to regulate lymphokine production, with a strong bias toward IFN-gamma and against IL-2 in the absence of these interactions. Our results have implications for the manipulation of T cell-dependent immune responses. PMID- 9862692 TI - Efficient lymphocyte migration across high endothelial venules of mouse Peyer's patches requires overlapping expression of L-selectin and beta7 integrin. AB - Lymphocyte migration into lymphoid organs is regulated by adhesion molecules including L-selectin and the beta7 integrins. L-selectin and alpha4beta7 are predominantly hypothesized to direct the selective migration of lymphocytes to peripheral lymph nodes and the gut-associated lymphoid tissues, respectively. To further characterize interactions between L-selectin and beta7 integrins during lymphocyte recirculation, mice deficient in both receptors (L-selectin/beta7 integrin-/-) were generated. The simultaneous loss of L-selectin and beta7 integrin expression prevented the majority of lymphocytes (>95% inhibition) from attaching to high endothelial venules (HEV) of Peyer's patches and other lymphoid tissues during in vitro binding assays. Moreover, the inability to bind HEV eliminated the vast majority of L-selectin/beta7 integrin-/- lymphocyte migration into Peyer's patches during short-term and long-term in vivo migration assays (>99% inhibition,p < 0.01). The lack of lymphocyte migration into Peyer's patches correlated directly with the dramatically reduced size and cellularity (99% reduced) of this tissue in L-selectin/beta7 integrin-/- mice. High numbers of injected L-selectin/beta7 integrin-/- lymphocytes remaining in the blood of wild type mice correlated with markedly increased numbers of circulating lymphocytes in L-selectin/beta7 integrin-/- mice. Loss of either L-selectin or the beta7 integrins alone resulted in significant but incomplete inhibition of Peyer's patch migration. Collectively, the phenotype of L-selectin/beta7 integrin-/- mice demonstrates that these two receptors primarily interact along the same adhesion pathway that is required for the vast majority of lymphocyte migration into Peyer's patches. PMID- 9862693 TI - Dependence of both spontaneous and antibody-dependent, granule exocytosis mediated NK cell cytotoxicity on extracellular signal-regulated kinases. AB - Extracellular signal-regulated kinases (ERK, also known as mitogen-activated protein kinases) are serine-threonine kinases transducing signals elicited upon ligand binding to several tyrosine kinase-associated receptors. We have reported that ERK2 phosphorylation and activation follows engagement of the low affinity receptor for the Fc portion of IgG (CD16) on NK cells, and is necessary for CD16 induced TNF-alpha mRNA expression. Here, we analyzed the involvement of ERK in NK cell-mediated cytotoxicity and IFN-gamma expression induced upon stimulation with targets cells, coated or not with Abs. Our data indicate that, as with immune complexes, ERK2 phosphorylation occurs in human primary NK cells upon interaction with target cells sensitive to granule exocytosis-mediated spontaneous cytotoxicity, and that this regulates both target cell- and immune complex induced cytotoxicity and IFN-gamma mRNA expression. A specific inhibitor of mitogen-activated protein kinase kinase reduced both spontaneous and Ab-dependent cytotoxicity in a dose-dependent manner involving, at least in part, inhibition of granule exocytosis without affecting effector/target cell interaction and rearrangement of the cytoskeleton proteins actin and tubulin. Involvement of ERK in the regulation of Ca2+-dependent cell-mediated cytotoxicity was confirmed, using a genetic approach, in primary NK cells infected with a recombinant vaccinia virus encoding an ERK inactive mutant. These data indicate that the biochemical pathways elicited in NK cells upon engagement of receptors responsible for either spontaneous or Ab-dependent recognition of target cells, although distinct, utilize ERK as one of their downstream molecules to regulate effector functions. PMID- 9862694 TI - Terminal deoxynucleotidyl transferase expression during neonatal life alters D(H) reading frame usage and Ig-receptor-dependent selection of V regions. AB - During neonatal life, Ig diversity is limited in many respects. The absence of terminal deoxynucleotidyl transferase (TdT) expression with the consequent lack of nontemplated addition during the neonatal period, coupled with the predominant usage of a single D(H) reading frame (RF), leads to severe limitations of diversity in the CDR3 region of Ig heavy (H) chains. The neonatal Ig H chain repertoire is also characterized by restricted V(H) usage, with predominant expression of certain V(H) segments, such as V(H)81x, that are rarely evident during adult life. In this report, we examine the effect of enforced TdT expression on the neonatal repertoire of V(H)81xDJ(H) rearrangements. We find that TdT synthesis abrogates D(H) RF bias during the fetal/neonatal period through a Ig-receptor-independent mechanism. These findings suggest that D(H) RF bias during neonatal life is determined largely by homology-directed joining. We also find that TdT synthesis alters the selection of productively rearranged V(H)81xDJ(H) alleles in the neonatal spleen through a Ig-receptor-dependent mechanism. Analysis of predicted CDR3 amino acid sequences indicates that positive selection of V(H)81x-encoded H chains is correlated with the presence of a consensus sequence immediately adjacent to the V(H) segment. These data support the hypothesis that the CDR3 region is critical in determining the ability of V(H)81x-encoded H chains to form functional receptors that support positive selection of B lymphocytes. Together, our results demonstrate that TdT can indirectly influence the Ig repertoire by influencing both receptor-dependent and receptor-independent selection processes. PMID- 9862695 TI - Transcriptional suppression of matrix metalloproteinase-2 gene expression in human astroglioma cells by TNF-alpha and IFN-gamma. AB - Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that function in the turnover of extracellular matrix components during development. In addition, MMPs also contribute to pathological conditions associated with inflammation, angiogenesis, and tumor invasion. A 72-kDa type IV collagenase, also referred to as gelatinase A or MMP-2, has been proposed to potentiate the invasion and metastasis of malignant tumors. In particular, MMP-2 activity has been shown to constitute an important component of human astroglioma invasion. We investigated the influence of various cytokines, both proinflammatory and immunosuppressive, on MMP-2 gene expression in two human astroglioma cell lines (U251-MG and CRT). Our results indicate that the cell lines constitutively express high levels of MMP-2 mRNA, protein, and bioactivity as assessed by ribonuclease protection assay, immunoblotting, and zymography assays, respectively. The proinflammatory cytokines TNF-alpha and IFN-gamma individually can inhibit constitutive MMP-2 expression, and function in an additive manner for near-complete inhibition of MMP-2 expression. Inhibition of MMP-2 mRNA levels by TNF-alpha and IFN-gamma is not due to destabilization of the MMP-2 message; rather, inhibition is mediated at the transcriptional level. Furthermore, TNF-alpha/IFN-gamma inhibition of MMP-2 expression results in decreased invasiveness of the human astroglioma cells through an extracellular matrix. These results raise the possibility that TNF-alpha and IFN-gamma may have beneficial effects in attenuating astroglioma invasive properties. PMID- 9862696 TI - Clonal expansion within CD4+ and CD8+ T cell subsets in human T lymphotropic virus type I-infected individuals. AB - To investigate the diversity of the T cell repertoire involved in human T lymphotropic virus type I (HTLV-I) infections, peripheral blood T cell subsets were analyzed by using a PCR-based assay that permits determination of complementarity-determining region 3 (CDR3) length variation in TCR Vbeta transcripts. In two of four asymptomatic HTLV-I carriers and in four of five patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), mono- or oligoclonal expansions were detected in the CD4+ T cell subset. In one patient with adult T cell leukemia, a specific clone bearing Vbeta7 was detected in the CD4+ T cell subset. In contrast, clonal expansion was not observed in the CD4 T cell subsets of three individuals with asymptomatic HTLV-II infection or in our previous studies of a large number of uninfected individuals. Oligoclonal expansions in the CD8+ T cell subset were detected in all subjects, including the patient with adult T cell leukemia. No differences in the number of expanded clones were noted between asymptomatic carriers and in patients with HAM/TSP and there was no obvious restriction in the TCR V region usage. Direct sequencing revealed no significant bias in the CDR3 motifs utilized by the predominant clones. This report is the first direct demonstration of clonal expansions within fractionated T cell subsets (CD4+ and CD8+) in HTLV-I infections and suggests that 1) clonal expansion of CD4+ T lymphocytes likely occurs as a direct result of infection and 2) polyclonal CD8+ T cell expansion occurs frequently and independently of disease association. PMID- 9862697 TI - Structural basis of the gp120 superantigen-binding site on human immunoglobulins. AB - B cell superantigens (SAg) interact with normal human nonimmune Igs (Igs), independently of the light chain isotype, and activate a large proportion of the B cell repertoire. Recently, the major envelope protein of HIV-1, gp120, was found to exhibit SAg-like properties for B cells with potential pathologic consequences for the infected host. This unconventional mode of interaction contrasts with its binding to immunization-induced Abs, which requires the tertiary structure of the heavy and light chain variable regions. In this report, we have examined the structural basis of the interaction between human Igs and gp120. We found that gp120 binding is restricted to Igs from the V(H)3 gene family and that the two V(H) genes 3-23 and 3-30, known to be overutilized during all stages of B cell development, frequently impart gp120 binding. We also provide evidence that the viral gp120 SAg can interact with only a subset of the human V(H)3+ Igs that can convey binding to the prototypic bacterial B cell SAg protein A from Staphylococcus aureus. Finally, we have identified amino acid positions present primarily in the first and third framework regions of the Ig heavy chain variable region, outside the conventional hypervariable loops, which correlate with gp120 binding. In a three-dimensional sequence-homology model, these residues partially overlap with the predicted SAg protein A binding site for V(H)3+ Igs. PMID- 9862698 TI - Redox regulation of caspase-3(-like) protease activity: regulatory roles of thioredoxin and cytochrome c. AB - Oxidative stress induces a variety of cellular responses, including apoptosis, and caspase family proteases are known to be involved in apoptosis. Caspase-3( like) protease activity was examined in Jurkat T cells to investigate the mechanism of apoptosis induced by a thioloxidant, diamide. Caspase-3 was activated when cells were cultured with 200 microM diamide that induced apoptosis, whereas no caspase-3 activation was detected with 500 microM diamide that induced necrosis. When apoptosis was induced in cells with exposure to 200 microM diamide, the intracellular thioredoxin (TRX) levels were maintained and the intracellular generation of reactive oxygen intermediates was marginal. The cytosolic fractions of cytochrome c were increased earlier than the activation of caspase-3. In contrast, when cells were exposed to 500 microM diamide, intracellular reactive oxygen intermediate generation was increased and processing of caspase-3 was not detected despite cytochrome c release, resulting in necrosis. Caspase-3 activity in cell lysate precultured with anti-Fas Ab was suppressed dose dependently by diamide and restored by thiol-reducing agents, DTT or TRX. When cells were precultured with 5 mM of buthionine sulfoximine, an inhibitor of glutathione synthesis, intracellular TRX levels were maintained, and as low as 20 microM diamide could induce apoptosis associated with the increase of cytosolic cytochrome c and the activation of caspase-3. These results indicate that the activation of caspase-3 in diamide-induced apoptosis is mediated, at least partly, by cytochrome c release from mitochondria, and the cellular reducing environment maintained by TRX, as well as glutathione, is required for caspase-3 activity to induce apoptosis. PMID- 9862699 TI - Production of a chimeric form of CD23 that is oligomeric and blocks IgE binding to the Fc epsilonRI. AB - The low affinity receptor for IgE (Fc epsilonRII/CD23) has previously been shown to interact with IgE with a dual affinity. Three chimeric constructs were created containing the lectin domain (amino acids 172-188) or the "neck" and lectin domain (amino acids 157-188) attached to subunits of oligomeric proteins. All chimeras were incapable of interacting with IgE with either a high or low affinity, indicating that the alpha-helical stalk of CD23 is important for orienting the lectin heads such that an interaction with IgE can occur. This concept received further support in that a chimeric CD23 composed of the human CD23 stalk and the mouse CD23 lectin head bound mouse IgE with a dual affinity, but could only bind rat IgE with a low affinity. Effort was next concentrated on a construct consisting of the entire extracellular (EC) region of CD23. A mutation to the first cleavage site of CD23 (C1M) resulted in a more stable molecule as determined by a decrease of soluble CD23 release. A soluble chimeric EC-C1M was prepared by attaching an isoleucine zipper to the amino terminus (lzEC C1M). The interaction with IgE by lzEC-C1M was found to be superior to that seen with EC-CD23. The lzEC-C1M could inhibit binding of IgE to both CD23 and the high affinity receptor for IgE, Fc epsilonRI, providing further evidence for a strong interaction with IgE. Fc epsilonRI inhibition (approximately 70%) was seen at equimolar concentrations of lzEC-C1M, implying the effectiveness of this chimera and suggesting its potential therapeutic value. PMID- 9862701 TI - Two families of GTPases dominate the complex cellular response to IFN-gamma. AB - IFN-gamma induces a number of cellular programs functional in innate and adaptive resistance to infectious pathogens. It has recently become clear that the complete cellular response to IFN-gamma is extraordinarily complex, with >500 genes (i.e., approximately 0.5% of the genome) activated. We made suppression subtractive hybridization differential libraries from IFN-gamma-stimulated primary mouse embryonic fibroblasts and from a mouse macrophage cell line, ANA-1, in each case with reference to unstimulated cells. Of approximately 250 clones sequenced at random from the two libraries, >35% were representatives of one or the other of two small unrelated families of GTPases, the 65-kDa and 47-kDa families. These families dominate the IFN-gamma-induced response in both cell types. We report here the full-length sequences of one new 65-kDa and two new 47 kDa family members. The 65-kDa family members are under transcriptional control of IRF-1, whereas the 47-kDa family members are inducible in embryonic fibroblasts from IRF-1(-/-) mice. Members of both GTPase families are strongly up regulated in livers of wild-type mice infected with the pathogenic bacterium, Listeria monocytogenes, but not in IFN-gammaR(0/0) mice. These GTPases appear to be dedicated to the IFN-gamma response, since resting levels are negligible and since neither family shows any significant relationship to any other described family of GTPases. Understanding the role of these GTPases in IFN-gamma-mediated resistance against pathogens is the task for the future. PMID- 9862702 TI - Marsupial light chains: complexity and conservation of lambda in the opossum Monodelphis domestica. AB - The Ig lambda chains in the South American opossum, Monodelphis domestica, were analyzed at the expressed cDNA and genomic organization level, the first described for a nonplacental mammal. The V lambda segment repertoire in the opossum was found to be comprised of at least three diverse V lambda families. Each of these families appears to be related to distinct V lambda families present in placental mammals, suggesting the divergence of these genes before the separation of metatherians and eutherians more than 100 million years ago. Based on framework and constant region sequences from full-length cDNAs and intron sequences from genomic clones, it appears that there are multiple functional J lambda-C lambda pairs in the opossum locus. The opossum J lambda-C lambda sequences are phylogenetically clustered, suggesting that these gene duplications are more recent and species specific. Sequence analysis of a large set of functional, expressed V lambda-J lambda recombinations is consistent with an unbiased, highly diverse lambda light chain repertoire in the adult opossum. Overall, the complexity of the Ig lambda locus appears to be greater than that found in the Ig heavy chain locus in the opossum, and light chains are therefore likely to contribute significantly to Ig diversity in this species. PMID- 9862700 TI - HLA (A*0201) mimicry by anti-idiotypic monoclonal antibodies. AB - Soluble MHC Ags and anti-Id (anti-anti-MHC) Abs have both been shown to inhibit MHC alloantigen-specific B cell responses in vivo. We hypothesized that some anti idiotypic Abs function as divalent molecular mimics of soluble HLA alloantigen. To test this idea, we studied two well-defined anti-idiotypic mAbs, T10-505 and T10-938, elicited in syngeneic BALB/c mice by immunization with CRll-351, an HLA A2,24,28-specific mAb. Each anti-Id induced "Ab-3" Abs in rabbits that cross reacted with HLA-A2 but not with HLA-B Ags. Furthermore, each anti-Id could bind to and block Ag recognition by Ha5C2.A2, a human homologue of mAb CRll-351. Both anti-Id mAb displayed weak reactivity with the human mAb SN66E3, which recognized an overlapping but distinct determinant of HLA-A2 Ags; neither reacted with human mAb MBW1, which recognized a nonoverlapping HLA-A2 determinant. Amino acid sequence comparison of mAb CRII-351 heavy and light chain variable region complementarity-determining regions (CDRs) with those of mAb Ha5C2.A2 and SN66E3 revealed short regions of homology with both human mAb; a large insert in the light chain CDR1 of mAb SN66E3 distinguished it from both CRll-351 and Ha5C2.A2. The amino acid sequences of mAb T10-505 and T10-938, which differed markedly from each other, revealed no homology to the alpha2 domain sequence of HLA-A*0201 that contains the CRll-351 mAb-defined epitope. We conclude that structurally different anti-Id Abs can mimic a polymorphic conformational epitope of an HLA Ag. In the case of T10-505 and T10-938 mimicry was not based on exact replication of the epitope by the hypervariable loops of the anti-Id mAb. PMID- 9862703 TI - MEK and ERK activation in ras-disabled RBL-2H3 mast cells and novel roles for geranylgeranylated and farnesylated proteins in Fc epsilonRI-mediated signaling. AB - Cross-linking the high affinity IgE receptor Fc epsilonRI of basophils and mast cells activates receptor-associated protein-tyrosine kinases and stimulates a signaling cascade leading to secretion, ruffling, spreading, and cytokine production. Previous evidence that the pan-prenylation inhibitor lovastatin blocks Ag-stimulated Ca2+ influx, secretion, and membrane/cytoskeletal responses implicated isoprenylated proteins in the Fc epsilonRI-coupled signaling cascade but could not distinguish between contributions of C15 (farnesylated) and C20 (geranylgeranylated) species. Here we establish concentrations of lovastatin and the farnesyl-specific inhibitor BZA-5B that inhibit the farnesylation and Ag induced activation of Ras species in RBL-2H3 cells (H-Ras, K-RasA, and K-RasB). These inhibitors have little effect on tyrosine kinase activation, which initiates Fc epsilonRI signaling. Although Ras is disabled, only lovastatin substantially blocks Raf-1 activation, and neither inhibitor affects mitogen activated protein kinase kinase/extracellular signal regulated kinase kinase (MEK) or ERK1/ERK2 activation. Thus, the pathway to Fc epsilonRI-mediated MEK/ERK and ERK activation can apparently bypass Ras and Raf-1. Predictably, only lovastatin inhibits Ag-induced ruffling, spreading, and secretion, previously linked to geranylgeranylated Rho and Rab family members. Additionally, only lovastatin inhibits phospholipase Cgamma-mediated inositol (1,4,5) trisphosphate production, sustained Ca2+ influx, and Ca2+-dependent IL-4 production, suggesting novel roles for geranylgeranylated (lovastatin-sensitive, BZA-5B-insensitive) proteins in Fc epsilonRI signal propagation. Remarkably, BZA-5B concentrations too low to inactivate Ras reduce the lag time to Ag-induced Ca2+ stores release and enhance secretion. These results link a non-Ras farnesylated protein(s) to the negative regulation of Ca2+ release from intracellular stores and secretion. We identified no clear role for Ras in Fc epsilonRI-coupled signaling but suggest its involvement in mast cell growth regulation based on the inhibition of cell proliferation by both BZA-5B and lovastatin. PMID- 9862704 TI - A region of conformational variability outside the peptide-binding site of a class I MHC molecule. AB - Peptide binding is known to influence the conformation of the surface of class I molecules as detected with mAbs and TCR. A new conformationally sensitive epitope on the mouse class I molecule Kb is defined by mAb AF6-88.5. The recognized structure is affected by amino acid substitutions in any of the three external domains of the class I heavy chain and, in addition, is influenced by the substitution of human for mouse beta2-microglobulin. Interestingly, the epitope for this Ab is not affected by mutations within the peptide-binding cleft or by the nature of the peptide bound. These findings indicate that the effect of a change in one domain of class I can radiate to other parts of the molecule. Furthermore, the existence of conformationally sensitive structures outside of the peptide-binding site suggests the possibility that class I molecules may change their structure in response to binding by receptors and ligands such as the TCR and the coligand CD8. Such structural changes may represent signals that can influence cellular activation events. PMID- 9862705 TI - CBF alpha3 (AML2) is induced by TGF-beta1 to bind and activate the mouse germline Ig alpha promoter. AB - TGF-beta1 directs class switching to IgA by splenic B cells and by the surface IgM+ B cell line, I.29mu, by inducing germline (GL) Ig alpha transcripts. The promoter segment between -130 and +46, relative to the first initiation site for mouse GL alpha transcripts, is sufficient for expression and TGF-beta1 inducibility of a reporter gene in B cell lines. Within this segment resides a TGF-beta1-responsive element (TbetaRE) that is required for induction of the promoter by TGF-beta1 and, when multimerized, is sufficient to transfer TGF-beta1 inducibility to another promoter. In this report we show that a TGF-beta1 inducible complex binds the TbetaRE and contains the transcription factor core binding factor (CBF; also known as acute myeloid leukemia, AML). Although all three CBF alpha family members activate the GL alpha promoter, only CBF alpha3 (AML-2) is induced by TGF-beta1 in splenic B and I.29mu cells. The TbetaRE contains two CBF binding sites. Mutation of both sites reduces but does not eliminate induction of the GL alpha promoter by TGF-beta1 or by overexpression of CBF, possibly due to the presence of an additional CBF site in the promoter. In addition, the TbetaRE contains two copies of another sequence motif. Mutation of these motifs eliminates TGF-beta1 induction of the GL alpha promoter. Together the data indicate that TGF-beta1 induction of the alpha promoter involves induction of CBF alpha3, which binds to the TbetaRE of the promoter along with one or more proteins. PMID- 9862707 TI - Analysis of signal transduction pathways regulating cytokine-mediated Fc receptor activation on human eosinophils. AB - Igs can be potent stimulants of eosinophil activation since interaction with IgA or IgG-coated particles can lead to eosinophil degranulation. We have investigated the comparative roles of mitogen-activated protein (MAP) kinases (MAPKs; ERK1/2 and p38) and phosphatidylinositol-3 kinase (PI3K) in the priming and regulation of Fc receptor functioning on human eosinophils utilizing a MAPK kinase (MEK) inhibitor (PD98059), a p38 inhibitor SB203580, and the widely used PI3K inhibitors wortmannin and LY294002. We demonstrate that priming of human eosinophils with Th2-derived cytokines, IL-4 and IL-5, differentially activate phosphotyrosine-associated PI3K and ERK and p38 MAP kinases. This activation can be inhibited by pre-incubation with wortmannin or LY294002, PD98059, and SB203580, respectively. Analysis of the effects of the inhibitors on rosette formation between human eosinophils and IgA- or IgG-coated beads revealed that activation of MEK was not required for IgA binding after priming with IL-4 or IL 5. However, inhibition of MEK did inhibit IL-5-primed binding of IgG-beads. The rosette formation of primed eosinophils with IgA-beads could be completely inhibited by wortmannin and LY294002 treatment, demonstrating a critical role for PI3K. Interestingly, inhibition of the p38 pathway also resulted in a complete blockade of IgA rosette formation. This work demonstrates regulatory control by inside-out signaling of Fc receptors by various cytokines on human eosinophils. Thus in vivo the local production of Th2-derived cytokines will regulate the effector functions of Fc receptors. PMID- 9862706 TI - Antitumor response elicited by a superantigen-transmembrane sequence fusion protein anchored onto tumor cells. AB - Superantigens stimulate T cells bearing certain TCR beta-chain variable regions when bound to MHC II molecules. We investigated whether the superantigen toxic shock syndrome toxin-1 (TSST1) could induce an antitumor immune response when anchored onto MHC II-negative tumor cells. Our approach was to facilitate association of TSST1 with cell membranes by fusing its coding region to the transmembrane region (TM) sequence of the proto-oncogene c-erb-B-2. TSST1-TM was expressed in bacteria with an N-terminal histidine tag and purified using nickel agarose affinity chromatography. Purified TSST1-TM added to cultures of several different MHC II-negative tumor cells spontaneously associated with cell membranes, as detected by flow cytometry. Because superantigens can direct cell mediated cytotoxicity against MHC II-positive cells, a TM fusion protein lacking the TSST1 MHC II binding domain (TSST(88-194)-TM) was also constructed. Tumor cells precoated with TSST1-TM or TSST(88-194)-TM stimulated proliferation of human peripheral blood lymphocytes in vitro whereas uncoated tumor cells did not. Mice preimmunized with TSST1-TM- or TSST(88-194)-TM-coated tumor cells mounted a systemic response that resulted in significant antitumor immunity as measured by regression of a parental tumor challenge. TSST1-TM and TSST(88-194)-TM fusion proteins represent a useful new strategy for attaching superantigens or potentially other proteins onto tumor cell surfaces without genetic manipulation. PMID- 9862708 TI - Resistance to the African trypanosomes is IFN-gamma dependent. AB - The role of variant surface glycoprotein (VSG)-specific Th cell responses in determining resistance to the African trypanosomes was examined by comparing Th cell responses in relatively resistant and susceptible mice as well as in cytokine gene knockout mice infected with Trypanosoma brucei rhodesiense. Resistant B10.BR and C57BL/6 mice expressed Th1 cell cytokine responses to VSG stimulation during infection, while susceptible C3H mice produced weak or no Th1 cell cytokine responses. Neither resistant B10.BR and C57BL/6 mice nor susceptible C3H mice made detectable Th2 cell cytokine responses to parasite Ag. To more closely examine the potential role of IFN-gamma and other cytokines in host resistance, we determined the resistance phenotypes and Th cell responses of IFN-gamma and IL-4 knockout mice. Infected C57BL/6-IFN-gamma knockout mice were as susceptible as C57BL/6-scid mice and made an IL-2, but not an IL-4, cytokine response to VSG, while C57BL/6-IL-4 knockout mice were as resistant as the wild type strain and exhibited both IL-2 and IFN-gamma cytokine responses. Passive transfer of spleen cells from wild-type mice to IFN-gamma knockout mice resulted in enhanced survival. Both wild-type and IFN-gamma knockout mice controlled parasitemia with VSG-specific Ab responses, although parasitemias were higher in the IFN-gamma knockout mice. Overall, this study demonstrates for the first time that relative resistance to African trypanosomes is associated with a strong Th1 cell response to parasite Ags, that IFN-gamma, but not IL-4, is linked to host resistance, and that susceptible animals do not make compensatory Th2 cell responses in the absence of Th1 cell cytokine responses. PMID- 9862709 TI - Coelomocytes express SpBf, a homologue of factor B, the second component in the sea urchin complement system. AB - A homologue of factor B, SpBf, has been cloned and sequenced from an LPS activated coelomocyte cDNA library from the purple sea urchin, Strongylocentrotus purpuratus. The deduced amino acid sequence and domain structure show significant similarity to the vertebrate Bf/C2 family proteins. SpBf is a mosaic protein, composed of five short consensus repeats, a von Willebrand Factor domain, and a serine protease domain. It has a deduced molecular mass of 91 kDa, with a conserved cleavage site for a putative factor D protease. It has ten consensus recognition sites for N-linked glycosylation. Amino acids involved in both Mg2+ binding and in serine protease activity in the vertebrate C2/Bf proteins are conserved in SpBf. Phylogenetic analysis of SpBf indicates that it is the most ancient member of the vertebrate Bf/C2 family. Additional phylogenetic analysis of the SCRs indicates that five SCRs in SpBf may be ancestral to three SCRs, which is the typical pattern in the vertebrate Bf/C2 proteins. RNA gel blots show that SpBf transcripts are 5.5 kb and are specifically expressed in coelomocytes. Genome blots suggest that the SpBf gene (Sp152) is single copy gene per haploid genome. This is the second complement component to be identified from the sea urchin, and, with the sea urchin C3 homologue, these two components may be part of a simple complement system that is homologous to the alternative pathway in higher vertebrates. PMID- 9862710 TI - Leishmania mexicana cysteine proteinase-deficient mutants have attenuated virulence for mice and potentiate a Th1 response. AB - Leishmania mexicana mutants lacking cysteine proteinase genes cpa (delta cpa), cpb (delta cpb), or both cpa and cpb (delta cpa/cpb) have been generated by targeted gene disruption. Delta cpa mutants produce a disease phenotype in BALB/c mice close to that of wild-type L. mexicana, but delta cpb mutants are much less infective, producing very slowly growing small lesions, and delta cpa/cpb double mutants do not induce lesion growth. Immunologic analysis of Ab isotype during infection and splenocyte IFN-gamma, IL-2, and IL-4 production following stimulation with Leishmania Ag or Con A indicates that there was a significant shift from a predominantly Th2-associated immune response in mice infected with wild-type L. mexicana to a Th1-associated response in mice inoculated with delta cpb or delta cpa/cpb. Significantly, delta cpa altered the balance of the immunologic response to a lesser extent than did the other mutants. Similar disease outcomes and switches in the Th1/Th2 balance were also observed when other L. mexicana-susceptible mouse strains were infected with the mutants. BALB/c and C57BL/6 mice vaccinated with delta cpa/cpb and CBA/Ca mice vaccinated with delta cpb or delta cpa/cpb were subsequently more resistant, to varying degrees, than were untreated mice to infection with wild-type parasites, as measured by development of lesions and parasite burden. These data implicate leishmanial cysteine proteinases not only as parasite virulence factors but also in modulation of the immune response and provide strong encouragement that cysteine proteinase-deficient L. mexicana mutants are candidate attenuated live vaccines. PMID- 9862711 TI - MRL/lpr and MRL+/+ macrophage DNA synthesis in the absence and the presence of colony-stimulating factor-1 and granulocyte-macrophage colony-stimulating factor. AB - Macrophage accumulation and proliferation as well as altered macrophage properties have been observed in autoimmune MRL mice. To determine whether there might be innate differences in the proliferative responses, we examined the DNA synthesis responses of peritoneal macrophages and macrophages derived in vitro from bone marrow precursors (bone marrow-derived macrophages (BMM)). Murine peritoneal exudate macrophages normally require the addition of macrophage CSF (CSF-1) to enter cell cycle in vitro. In contrast, we have found that many thioglycollate-induced adherent peritoneal macrophages, but not resident peritoneal macrophages, from both MRL/lpr and MRL+/+ mice atypically underwent DNA synthesis even in the absence of added CSF-1. They also responded very well to granulocyte-macrophage CSF. These findings may help to explain the appearance of increased macrophage numbers in MRL lesions. In contrast to a previous report, it was found that MRL/lpr and MRL+/+ BMM did not have an enhanced response to CSF 1 and that modulation of CSF-1 receptor expression was not more rapid in MRL BMM. We also found no evidence for abnormal CSF-1 internalization and degradation or for the lpr mutation to have any enhanced effect on BMM survival in the absence of CSF-1. TNF-alpha lowered the DNA synthesis response to CSF-1 of MRL/lpr BMM rather than enhanced it, as has been reported. Our data suggest that the enhanced accumulation of macrophages in the MRL/lpr kidney cannot be explained by a proposed model of enhanced responsiveness of MRL/lpr BMM to CSF-1, including a contribution by TNF-alpha. PMID- 9862712 TI - Alteration of intracellular calcium flux and impairment of nuclear factor-AT translocation in T cells during acute Toxoplasma gondii infection in mice. AB - Down-regulation of host immune response to Toxoplasma gondii is associated with the expression of specific cytokines, in particular IL-10, and the induction of CD4+ T cell anergy. In the present study we report that the expression of both CD4 and CD2 antigen is down-regulated during the acute phase of infection. A decrease in the expression of CD2 was apparent during the acute phase of T. gondii infection in three genetically distinct strains of mice, CBA/J, C57BL/6, and BALB/c. The lymphoproliferative response induced by cross-linked anti-CD3 mAb or by Con A was markedly depressed. This suppressed response was associated with a reduction in the influx of Ca2+. We have examined whether lymphocytes from T. gondii mice maintain NF-AT transcription factors in the nucleus where they participate in the Ca2+-dependent induction of genes required for lymphocyte activation and proliferation. Immunofluorescence with confocal microscopy using an Ab to NF-ATc demonstrates a decrease in translocation of NF-ATc in T lymphocytes from acutely infected mice. Together, these results suggest that the defect in T cell expansion that occurs during acute murine toxoplasmosis is related to reduced activity of NF-AT, a calcium-dependent transcription factor required for T cell proliferation. PMID- 9862713 TI - Expression of the third component of complement, C3, in regenerating limb blastema cells of urodeles. AB - In this study we have shown that complement component C3 is expressed in the regenerating tissue during urodele limb regeneration. C3 was expressed in the dedifferentiated regeneration blastema and in the redifferentiated limb tissues in the axolotl, Amblystoma mexicanum, and in Notophthalmus viridescens. This expression was verified by immunofluorescent staining using an Ab against axolotl C3 and by in situ hybridization with an axolotl C3 cDNA probe. In the early stages of regeneration C3 appeared to be equally present in all mesenchymal cells and in the wound epithelium, whereas in the later stages it was mainly expressed in the differentiating muscle cells. Since no expression was seen in the developing limb, it appears that the C3 expression was specific to the regeneration process. We then demonstrated by hybridization experiments that a blastema cell line of myogenic origin expresses C3. All these findings implicate C3 in the dedifferentiation process and may indicate a new role for this molecule in muscle differentiation. PMID- 9862715 TI - Contribution of dermal macrophage trafficking in the sensitization phase of contact hypersensitivity. AB - We investigated cellular trafficking of dermal macrophages that express a macrophage calcium-type lectin (MMGL) during the sensitization of delayed-type hypersensitivity. In skin, dermal macrophages, but not epidermal Langerhans cells, have been shown to express MMGL. Epicutaneous sensitization by FITC produced a transient increase in MMGL-positive cells in regional lymph nodes. To directly investigate whether the increase was due to cell migration from dermis, MMGL-positive cells purified from skin were intradermally injected into syngeneic mice after labeling with a fluorescent cell tracer, followed by epicutaneous sensitization over the site of injection. MMGL-positive cells containing the tracer were found in the regional lymph nodes after sensitization. The majority of the MMGL-positive cell migrants were negative for FITC fluorescence despite the presence of FITC-labeled cells that included Langerhans cell migrants. Because the extent of MMGL-positive cell migration was greatly influenced by the selection of vehicles to dissolve FITC, the efficiency of sensitization was compared using the ear swelling test. Migration of both Langerhans cells (FITC labeled cells) and MMGL-positive cells contributed positively to the efficiency of sensitization. Interestingly, MMGL-positive cell migration was induced by vehicle alone, even in the absence of FITC. These results suggest that migration of dermal MMGL-positive cells accounts for the adjuvant effects of vehicles at least in part. PMID- 9862714 TI - Interaction of very late antigen-4 with VCAM-1 supports transendothelial chemotaxis of monocytes by facilitating lateral migration. AB - The transient regulation of very late antigen (VLA)-4 avidity by CC chemokines may promote chemotaxis of monocytes across VCAM-1-bearing barriers, whereas late and prolonged activation of VLA-5 may mediate subsequent localization in the extracellular matrix. We demonstrate that interactions of VLA-4 with VCAM-1, fibronectin, or a 40-kDa fragment but not a 120-kDa fragment of fibronectin supported the lateral random migration of isolated blood monocytes induced by CC chemokines, termed chemokinesis. This effect was optimal at intermediate substrate concentrations. Moreover, coimmobilization of VCAM-1 with ICAM-1 allowed better migration than ICAM-1 alone. Chemokinesis on VCAM-1 appeared to be associated with transient regulation of VLA-4 avidity by CC chemokines, given that locking VLA-4 in a high avidity state markedly inhibited migration and the locomotion rate was inversely correlated with the adhesive strength of VLA-4 to VCAM-1 following stimulation with monocyte chemoattractant protein-1. Induction of VCAM-1 expression by endothelial activation with IL-4 improved chemokinesis and lateral migration toward a monocyte chemoattractant protein-1 or a monocyte inflammatory protein-1alpha gradient on endothelium and increased transendothelial chemotaxis of monocytes by a VLA-4-dependent mechanism. In contrast, endothelial activation with IL-4 did not affect the time required for diapedesis of monocytes itself. Hence, VCAM-1 may facilitate transendothelial chemotaxis by supporting lateral migration of attached monocytes along endothelium. PMID- 9862716 TI - Matrix metalloproteinases generate angiostatin: effects on neovascularization. AB - Angiostatin, a cleavage product of plasminogen, has been shown to inhibit endothelial cell proliferation and metastatic tumor cell growth. Recently, the production of angiostatin has been correlated with tumor-associated macrophage production of elastolytic metalloproteinases in a murine model of Lewis lung cell carcinoma. In this report we demonstrate that purified murine and human matrix metalloproteinases generate biologically functional angiostatin from plasminogen. Macrophage elastase (MMP-12 or MME) proved to be the most efficient angiostatin producing MMP. MME was followed by gelatinases and then the stomelysins in catalytic efficiency; interstitial collagenases had little capacity to generate angiostatin. Both recombinant angiostatin and angiostatin generated from recombinant MME-treated plasminogen inhibited human microvascular endothelial cell proliferation and differentiation in vitro. Finally, employing macrophages isolated from MME-deficient mice and their wild-type littermates, we demonstrate that MME is required for the generation of angiostatin that inhibits the proliferation of human microvascular endothelial cells. PMID- 9862717 TI - Lymphotoxin alpha3 induces chemokines and adhesion molecules: insight into the role of LT alpha in inflammation and lymphoid organ development. AB - Lymphotoxin (LT) plays an important role in inflammation and lymphoid organ development, though the mechanisms by which it promotes these processes are poorly understood. Toward this end, the biologic activities of a recently generated recombinant murine (m) LT alpha preparation were evaluated. This cytokine preparation was effective at inducing cytotoxicity of WEHI target cells with 50% maximal killing observed with 1.2 ng/ml. mLT alpha also induced the expression of inflammatory mediators in the murine endothelial cell line bEnd.3. rmLT alpha induced expression of the adhesion molecules VCAM, ICAM, E-selectin, and the mucosal addressin cellular adhesion molecule, MAdCAM-1. When mLT alpha, human (h) LT alpha, and mTNF-alpha were compared, mLT alpha was the most potent inducer of MAdCAM-1. None of these cytokines induced the peripheral node addressin, PNAd. mLT alpha also induced expression of the chemokines RANTES, IFN inducible protein 10 (IP-10), and monocyte chemotactic protein 1 (MCP-1). mRNA levels peaked 4 h following treatment with mLT alpha and declined through the 24 h treatment period. LT alpha also induced chemokine protein within 8 h of treatment, which increased through the 24-h treatment period. These data demonstrate that the proinflammatory effects of LT alpha3 may be mediated in part through the induction of adhesion molecule and chemokine expression. Further, LT alpha3 may promote development of lymphoid tissue through induction of chemokines and the mucosal addressin MAdCAM-1. These data confirm previous observations in transgenic and knockout mice that LT alpha3 in the absence of LT beta carries out unique biologic activities. PMID- 9862718 TI - Lipopolysaccharide-induced leukocyte rolling and adhesion in the rat mesenteric microcirculation: regulation by glucocorticoids and role of cytokines. AB - A common side effect of high dose glucocorticoid therapy is increased susceptibility to bacterial infection, an effect that is in part mediated through inhibition of leukocyte recruitment to infected areas. However, the sites at which glucocorticoids act to prevent the multistep process of leukocyte recruitment have not been fully established. In this study, the effects of the glucocorticoid dexamethasone (DEX) on leukocyte-endothelial interactions, in response to bacterial LPS, were examined utilizing a model of rat mesenteric intravital microscopy. Pretreatment of rats with DEX (0.5 mg/kg) for 18 h or 30 min before stimulation with LPS significantly inhibited LPS-induced leukocyte rolling and adhesion in mesenteric postcapillary venules. Pretreatment with DEX also inhibited LPS-induced changes in expression of L-selectin and a shared epitope of CD11b/c on circulating neutrophils. These effects of DEX may be due to DEX inhibition of IL-1, TNF, and cytokine-induced neutrophil chemoattractant-1 (CINC-1) generation, since antagonists to these mediators were able to mimic DEX effects on leukocyte-endothelial interactions and circulating leukocyte phenotype. These data indicate that inhibition of cytokine- and chemokine-induced leukocyte-endothelial interactions may be a primary mechanism by which glucocorticoids inhibit leukocyte recruitment to bacterial agents and thus increase susceptibility to infection. PMID- 9862719 TI - The type II IL-1 receptor interacts with the IL-1 receptor accessory protein: a novel mechanism of regulation of IL-1 responsiveness. AB - IL-1 binds to two types of receptors on the cell membrane, of which only type I (IL-1RI) transduces signals in concert with the coreceptor IL-1 receptor accessory protein (IL-1RAcP) while type II (IL-1RII) allegedly functions solely as ligand sink and decoy receptor without participating in IL-1 signaling. To investigate the regulatory role of IL-1RII on IL-1 responsiveness, a chimeric receptor encompassing the extracellular and transmembrane portions of IL-1RII and the cytoplasmic signal-transducing domain of IL-1RI was transfected into two murine EL-4-derived sublines that do or do not express IL-1RAcP, respectively. The chimeric receptor was able to transduce the IL-1 signal and induce IL-2 production only in the cell line which expressed IL-1RAcP, suggesting effective interaction between the extracellular domains of IL-1RII and IL-1RAcP in the presence of IL-1. The physical association of ligated IL-1RII with IL-1RAcP was proven by crosslinking experiments with radio-iodinated IL-1 and subsequent immunoprecipitations in normal human B cells and in EL-4 D6/76 cells transiently cotransfected with IL-1RII and IL-1RAcP, respectively. Based on these findings, it is proposed that upon IL-1 binding IL-1RII can recruit IL-1RAcP into a nonfunctional trimeric complex and thus modulate IL-1 signaling by subtracting the coreceptor molecule from the signaling IL-1RI. In this novel mechanism of coreceptor competition, the ratio between IL-1RII and IL-1RI becomes the central factor in determining the IL-1 responsiveness of a cell and the availability of IL-1RAcP becomes limiting for effective IL-1 signaling. PMID- 9862720 TI - Intrinsic defects in macrophage IL-12 production associated with immune dysfunction in the MRL/++ and New Zealand Black/White F1 lupus-prone mice and the Leishmania major-susceptible BALB/c strain. AB - We have demonstrated that macrophages (Mphi) from young, prediseased, lupus-prone MRL/++ and New Zealand Black/White F1 mice display defective production of TNF alpha, IL-1, and IL-6, but normal production of IL-10. In an attempt to determine the potential functional implications of this phenotype for autoimmunity, we demonstrate here that endotoxin-activated Mphi from these lupus-prone mice showed dramatically reduced expression of IL-12, a cytokine essential for Th1 responses that may be defective during lupus. IL-12 production was also reduced by Mphi from the control BALB/c strain, compatible with the concept that a genetically programmed deficit in IL-12 levels may underlie the IL-4-dominated BALB/c response to infection by the parasite Leishmania major. Although both IL-12 and TNF-alpha expression defects by Mphi from lupus-prone strains are expressed rapidly after activation, treatment with each cytokine demonstrated that only TNF alpha contributes to the subsequent dysregulation of Mphi IL-1 and IL-6 expression in these strains, and that the reduced autocrine activity of defective IL-12 or TNF-alpha levels was not causal to each other. Although the intrinsic defect in IL-12 expression by lupus-prone and BALB/c Mphi may lead to defective Th1 responses, these Mphi responded to the Th1-derived cytokine, IFN-gamma, in a normal fashion suggesting a defective role in the induction, rather than the propagation, of Th1 responses in these mice. Our finding of a conserved intrinsic defect in IL-12 production by Mphi from the two principal mouse models of multigenic lupus provides insight into how excessive humoral responses may develop, and perhaps be prevented, in systemic autoimmune disease. PMID- 9862721 TI - Isolation and characterization of a variant HL60 cell line defective in the activation of the NADPH oxidase by phorbol myristate acetate. AB - Promyelocytic human leukemia HL60 cells can be differentiated into neutrophil like cells that exhibit an NADPH oxidase activity through direct stimulation of protein kinase C (PKC) with PMA or through formyl peptide receptor activation. We have isolated a variant HL60 clone that exhibited a conditional PMA-induced oxidative response depending on the agent used for the differentiation. While cells differentiated with DMSO responded to either PMA or N-formyl peptide (N formyl-Met-Leu-Phe-Lys or fMLFK), cells differentiated with dibutyryl-cAMP (Bt2cAMP) responded to fMLFK but very poorly to PMA. However, in Bt2cAMP differentiated cells, the expression of the different PKC isoforms was similar to that observed in DMSO-differentiated cells. Moreover, PMA was able to induce a normal phosphorylation of the cytosolic factor p47phox and to fully activate extracellular signal-regulated kinases (Erk1/2). Interestingly, Bt2cAMP differentiated cells exhibited a strong and sustained O2- production when costimulated with PMA and suboptimal concentrations of fMLFK which were, per se, ineffective. This sustained response was only slightly reduced by the conjunction of the mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor PD98059 and wortmannin, a phosphatidylinositol-3 kinase (PI3K) inhibitor. Variant HL60 cells that were stably transfected with a constitutively active form of Rac1 were able, when differentiated with Bt2cAMP, to secrete oxidant following PMA stimulation. Altogether, the results suggest that, in addition to the phosphorylation of p47phox, the activation of NADPH oxidase requires the activation of a Rac protein through a pathway that diverges at a point upstream of MEK and that is independent of the activation of wortmannin sensitive PI3K. PMID- 9862723 TI - Successful induction of adjuvant arthritis in mice by treatment with a monoclonal antibody against IL-4. AB - Adjuvant arthritis (AA) is an experimental model of autoimmune disease in rats induced by immunization with Mycobacterium tuberculosis (MT). Induction of AA in other species, including mice, has been shown to be difficult. In the present study, we found that AA could be induced in mice if the animals were treated with a mAb (11B11 mAb) against IL-4. Histologically, the joints exhibited synovial edema with infiltration of many neutrophils in the early phase of inflammation. In its late phase, there were proliferation of synovium, cell infiltrate in which mononuclear cells predominated, and destruction of cartilage and subchondral bone. The joint inflammation was passively transferred to normal syngeneic recipient mice with lymphoid cells but not with sera from mice immunized with MT followed by treatment with the anti-IL-4 Ab. Delayed-type hypersensitivity (DTH) and proliferative responses of lymphoid cells to purified protein derivative were markedly augmented in 11B11 mAb-treated mice. Furthermore, the induction of arthritis was associated with a marked decrease in IL-4 secretion but a significant increase in IFN-gamma and IL-2 production. Thus, the neutralization of IL-4 by an anti-IL-4 Ab appears to be required for the induction of AA in mice. PMID- 9862722 TI - Astrocyte-derived monocyte-chemoattractant protein-1 directs the transmigration of leukocytes across a model of the human blood-brain barrier. AB - The migration of leukocytes across the blood-brain barrier (BBB) into the central nervous system is critical in the pathogenesis of central nervous system inflammatory diseases. The production of chemokines, such as monocyte chemoattractant protein-1 (MCP-1), by endothelial cells (EC) and astrocytes may initiate and amplify this process. Using a coculture of human EC and astrocytes to model the BBB, we demonstrated that exogenous MCP-1 induces the transmigration of monocytes in a dose-dependent manner. TNF-alpha, IFN-gamma, or IL-1beta treatment of cocultures also induced significant migration of monocytes that correlates with the induction of MCP-1 protein. TGF-beta, previously shown to induce MCP-1 expression in astrocytes, but not in EC, caused migration of monocytes across cocultures, but not across EC grown alone. Monocytes and lymphocytes transmigrated across cytokine-treated cocultures in greater numbers than across EC alone. Astrocytes were the main source of cytokine-induced MCP-1, supporting a role for astrocytes in facilitating leukocyte transmigration. A blocking Ab to MCP-1 inhibited MCP-1- and cytokine-induced transmigration of monocytes by 85-90%. Cytokine treatment of cocultures also resulted in the transmigration of activated, CD69-positive lymphocytes. The MCP-1-mediated transmigration of monocytes across cocultures was blocked using an Ab to ICAM-1 and inhibited by 55% using an Ab to E-selectin. These data suggest a central role for astrocyte-derived MCP-1 in directing the migration of monocytes and lymphocytes across the BBB. PMID- 9862724 TI - Antineutrophil cytoplasmic antibodies preferentially engage Fc gammaRIIIb on human neutrophils. AB - Antineutrophil cytoplasmic Abs (ANCA) are found in the circulation of many patients with systemic vasculitis. ANCA bind to ANCA target, such as proteinase 3 and myeloperoxidase, and activate neutrophils in an Fc gammaR-dependent manner. Human neutrophils constitutively express Fc gammaRIIa (CD32) and Fc gammaRIIIb (CD16), and there is clear in vitro experimental evidence of ANCA-mediated engagement of Fc gammaRIIa. However, direct experimental evidence of ANCA engagement of neutrophil Fc gammaRIIIb has been obscured by technical problems related to activation-induced receptor shedding and activation-induced expression of receptor on the surface of neutrophils. In this study, by blocking receptor shedding and using appropriate reporter anti-Fc gammaR mAb, we show that human cANCA and pANCA, and murine mAb with corresponding reactivities, can indeed engage Fc gammaRIIIb. Furthermore, our data suggest that Fc gammaRIIIb is preferentially engaged by ANCA relative to Fc gammaRIIa presumably due to the nearly 10-fold excess of Fc gammaRIIIb expression relative to Fc gammaRIIa expression. These results clearly demonstrate that the Fc region of ANCA bound to an ANCA target on the neutrophil surface engage Fc gammaRIIIb and indicate that Fc gammaRIIIb and Fc gammaRIIa may both be active participants in ANCA-induced neutrophil activation. However, given the low levels of ANCA target expression on neutrophils from patients with systemic vasculitis, Fc gammaRIIIb is likely to play a critical role in initiating and perpetuating ANCA-induced neutrophil activation. PMID- 9862725 TI - IL-4 inhibits mouse mast cell Fc epsilonRI expression through a STAT6-dependent mechanism. AB - Mast cell activation by IgE-mediated stimuli is a central event in atopic disease. The regulation of the mast cell high affinity receptor, Fc epsilonRI, is poorly understood. We show that IL-4 can inhibit Fc epsilonRI expression on mouse bone marrow-derived mast cells and fetal liver-derived mast cell progenitors. This effect could be observed at 2.5 ng/ml IL-4 and was dose dependent. IL-4 mediated inhibition of cultured BMMC required 4 days of stimulation and was sustained at maximum levels for at least 21 days. The inhibition of Fc epsilonRI expression resulted in decreased sensitivity to IgE-mediated stimulation, as measured by serotonin release, and the induction of mRNA for IL-4, IL-5, IL-6, and IL-13. Additionally, IL-4 could abrogate the IgE-mediated increase in Fc epsilonRI expression. Lastly, IL-4-mediated inhibition was dependent upon expression of the STAT6 transcription factor, as STAT6-deficient bone marrow derived mast cells did not decrease Fc epsilonRI levels in response to IL-4. These data argue for a homeostatic role of IL-4 in the regulation of Fc epsilonRI expression, a role that could be critical to understanding atopic disease. PMID- 9862726 TI - The first subcomponent of complement, C1q, triggers the production of IL-8, IL-6, and monocyte chemoattractant peptide-1 by human umbilical vein endothelial cells. AB - We and others have demonstrated previously the occurrence of cC1qR/CaR, a receptor for the collagen-like stalks of complement component C1q, on endothelial cells. In the present study we investigated whether binding of C1q to endothelial cells resulted in enhancement of cytokine or chemokine production. HUVEC produced 82 +/- 91 pg/ml of IL-8, 79 +/- 113 pg/ml of IL-6, and 503 +/- 221 pg/ml of monocyte chemoattractant peptide-1 (MCP-1) under basal conditions. Incubation with C1q resulted in a time- and dose-dependent up-regulation of IL-8 (1012 +/- 43 pg/ml), IL-6 (392 +/- 20 pg/ml), and MCP-1 (2450 +/- 101 pg/ml). This production is dependent on de novo protein synthesis, as demonstrated by the detection of specific mRNA after C1q stimulation, and inhibition of peptide production in the presence of cycloheximide. The production of all factors was inhibited (69 +/- 7%) by the collagenous fragments of C1q, while the C1q globular heads only induced 13 +/- 11% inhibition. When HUVEC were incubated with C1q in the presence of aggregated IgM, enhanced production of IL-8 (2500 +/- 422 pg/ml), IL-6 (997 +/- 21 pg/ml), and MCP-1 (5343 +/- 302 pg/ml) was found. Furthermore, F(ab')2 anti-calreticulin partially inhibited the production of IL-8, confirming at least the involvement of cC1qR/CaR. These experiments suggest that in an inflammatory response C1q not only is able to activate the complement pathway, but when presented in a proper fashion also might induce the production of factors that contribute to acute phase responses and recruitment of inflammatory cells. PMID- 9862727 TI - The role of adhesion molecules in human leukocyte attachment to porcine vascular endothelium: implications for xenotransplantation. AB - Many obstacles still prevent successful xenotransplantation of porcine donor organs. When hyperacute rejection is averted, transplanted pig organs are subject to acute vascular and cellular rejection. In autologous systems, leukocyte recruitment into inflamed tissues involves selectins, integrins, and Ig family members. To determine whether these mechanisms allow human leukocytes to effectively enter porcine grafts, the pathways by which human leukocytes adhere to TNF-alpha-stimulated porcine aortic endothelium were examined under static and physiologic flow conditions. L-selectin and E-selectin had overlapping functions in neutrophil capture and rolling, whereas Ab blockade of E-selectin and the beta2 integrins inhibited firm arrest of rolling neutrophils. Combined blockade of selectins and beta2 integrins resulted in negligible human neutrophil attachment to pig endothelium. Lymphocyte attachment to porcine endothelium was primarily L-selectin mediated, whereas beta2 integrin and VCAM-1/very late Ag-4 (VLA-4) interactions promoted static adhesion. Concurrent beta2 integrin, VLA-4, VCAM-1, and L-selectin blockade completely inhibited lymphocyte attachment. Thus, interactions between leukocyte-endothelial cell adhesion receptor pairs remained remarkably intact across the human-porcine species barrier. Moreover, disrupting the adhesion cascade may impair the ability of human leukocytes to infiltrate a transplanted porcine organ during rejection. PMID- 9862728 TI - Peptide modification or blocking of CD8, resulting in weak TCR signaling, can activate CTL for Fas- but not perforin-dependent cytotoxicity or cytokine production. AB - This study describes a form of partial agonism for a CD8+ CTL clone, S15, in which perforin-dependent killing and IFN-gamma production were lost but Fas (APO1 or CD95)-dependent cytotoxicity preserved. Cloned S15 CTL are H-2Kd restricted and specific for a photoreactive derivative of the Plasmodium berghei circumsporozoite peptide PbCS 252-260 (SYIPSAEKI). The presence of a photoactivatable group in the epitope permitted assessment of TCR-ligand binding by TCR photoaffinity labeling. Selective activation of Fas-dependent killing was observed for a peptide-derivative variant containing a modified photoreactive group. A similar functional response was obtained after binding of the wild-type peptide derivative upon blocking of CD8 participation in TCR-ligand binding. The epitope modification or blocking of CD8 resulted in an > or = 8-fold decrease in TCR-ligand binding. In both cases, phosphorylation of zeta-chain and ZAP-70, as well as calcium mobilization were reduced close to background levels, indicating that activation of Fas-dependent cytotoxicity required weaker TCR signaling than activation of perforin-dependent killing or IFN-gamma production. Consistent with this, we observed that depletion of the protein tyrosine kinase p56(lck) by preincubation of S15 CTL with herbimycin A severely impaired perforin- but not Fas-dependent cytotoxicity. Together with the observation that S15 CTL constitutively express Fas ligand, these results indicate that TCR signaling too weak to elicit perforin-dependent cytotoxicity or cytokine production can induce Fas-dependent cytotoxicity, possibly by translocation of preformed Fas ligand to the cell surface. PMID- 9862729 TI - Activation of phosphatidylinositol 3-kinase, protein kinase B, and p70 S6 kinases in lipopolysaccharide-stimulated Raw 264.7 cells: differential effects of rapamycin, Ly294002, and wortmannin on nitric oxide production. AB - Phosphatidylinositol 3-kinase (PI 3-kinase) and protein kinase B are critical players in cell proliferation and survival. Their downstream effector protein kinase, p70 S6 kinase, has an established role in protein translation. The mechanism by which bacterial LPS induces production of nitric oxide (NO) in murine macrophages is incompletely understood, and a role for PI 3-kinase/p70 S6 kinase pathway had not been previously investigated. In this study we demonstrate that LPS induced a fivefold activation of p70 S6 kinase and a twofold stimulation of PI 3-kinase. Pretreatment of Raw 264.7 cells with either rapamycin or Ly290042 completely blocked LPS-induced activation of p70 S6 kinase. Protein kinase B was also activated (twofold) by LPS and was only minimally affected by these inhibitors. PI 3-kinase activity was inhibited by both Ly294002 and wortmannin. The effects on NO production by these agents were strikingly different. While both rapamycin and Ly294002 resulted in almost complete inhibition of NO production, wortmannin was ineffective. Surprisingly, none of the inhibitors reduced the production of the inducible nitric oxide synthase protein (iNOS) as determined by immunoprecipitation. In vivo labeling studies revealed that the iNOS protein was phosphorylated in concordance with the production of NO. We conclude that LPS-mediated NO production occurs via a PI 3-kinase-independent, but FKBP12-rapamycin-associated protein-dependent, pathway in RAW cells by a mechanism probably involving phosphorylation of iNOS. PMID- 9862730 TI - Diversity of the fine specificity displayed by HLA-A*0201-restricted CTL specific for the immunodominant Melan-A/MART-1 antigenic peptide. AB - HLA-A*0201 melanoma patients often develop a CTL response to an immunodominant peptide derived from the melanocyte lineage-specific protein Melan-A/MART-1. We have shown previously that the antigenic peptide most often involved is the decapeptide Melan-A(26-35) (EAAGIGILTV). We also observed some clonal diversity in the fine specificity of Melan-A-specific CTL. To substantiate this observation, we have now tested a series of Melan-A(26-35) variant peptides containing single alanine substitutions for binding to HLA-A*0201 and recognition by polyclonal and monoclonal Melan-A-specific CTL. Substitution of several residues with alanine reduced peptide binding activity by > 10-fold. In contrast, substitution of E26 with alanine (AAAGIGILTV) resulted in a 5-fold higher binding activity as well as in stronger stability of the corresponding HLA-A*0201/peptide complexes. Interestingly, the peptide variant AAAGIGILTV was recognized more efficiently than the natural decapeptide by short term cultured, tumor infiltrated lymph node cell cultures and a number of Melan-A-specific CTL clones derived from different individuals. Moreover, this analysis revealed that the fine specificity of the CTL response to the Melan-A immunodominant epitope is quite diverse at the clonal level. At least three distinct patterns of fine specificity were identified. This diversity appears to reflect the diversity of the TCR repertoire available for this Ag, since similar results were obtained with a panel of Melan-A-specific CTL clones derived from a single melanoma patient. These findings have important implications for the formulation of Melan A peptide-based vaccines as well as for the monitoring of Melan-A-specific CTL responses in melanoma patients. PMID- 9862731 TI - Early development and spreading of autoantibodies to epitopes of IA-2 and their association with progression to type 1 diabetes. AB - Autoimmunity precedes clinical type 1 diabetes, and indicators of maturing autoimmune responses may be useful markers for disease prediction. To study this, epitope maturation of autoantibodies to the related protein tyrosine phosphatase (PTP)-like autoantigens IA-2 and IA-2beta was examined in sequential samples from birth in a cohort of 21 offspring developing multiple islet autoantibodies and a similar cohort of 48 relatives of patients with type 1 diabetes recruited at an older age. Initial reactivity in offspring was heterogeneous against the IA-2 juxtamembrane region (10/21) and PTP domains (13/21), and both specificity and extent of initial IA-2/IA-2beta autoantibodies were associated with HLA class II genotype. Intra-IA-2 epitope spreading and/or intermolecular spreading to IA 2beta epitopes were observed in seven offspring. In contrast, in older relatives, IA-2/IA-2beta Ab reactivity was stable and spreading rare. Development of diabetes in children was associated with the presence of Abs to the IA-2 juxtamembrane region (risk by age 5 yr, 52% vs 0% in those with PTP domain Abs only; p < 0.02), and 5 of 26 relatives who developed diabetes had IA-2 Abs only against the juxtamembrane region. The findings show that autoantibody reactivity to IA-2/IA-2beta is dynamic in the young, show that the juxtamembrane region of IA-2 is an early IA-2 autoantibody target, and suggest that these Abs are a risk factor for development of type 1 diabetes in infancy. PMID- 9862732 TI - HLA-independent heterogeneity of CD8+ T cell responses to MAGE-3, Melan-A/MART-1, gp100, tyrosinase, MC1R, and TRP-2 in vaccine-treated melanoma patients. AB - An important element in melanoma vaccine construction is to identify peptides from melanoma-associated Ags that have immunogenic potential in humans and are recognized by CD8+ T cells in vivo. To identify such peptides, we evaluated HLA A*02+ melanoma patients immunized to a polyvalent vaccine containing multiple Ags, including MAGE-3, Melan-A/MART-1, gp100, tyrosinase, melanocortin receptor (MC1R), and dopachrome tautomerase (TRP-2). Using a filter spot assay, we measured peripheral blood CD8+ T cell responses, before and after immunization, to a panel of 45 HLA-A*0201-restricted peptides derived from these Ags. The peptides were selected for immunogenic potential based on their strong binding affinity in vitro to HLA-A*0201. Vaccine treatment induced peptide-specific CD8+ T cell responses to 22 (47.8%) of the peptides. The most striking finding was the HLA-independent heterogeneity of responses to both peptides and Ags. All responding patients reacted to different combination of peptides and Ags even though the responding patients were all A*0201+ and the peptides were all A*0201 restricted. From 9 to 27% of patients developed a CD8+ T cell response to at least one peptide from each Ag, but no more than 3 (14%) reacted to the same peptide from the same Ag. This heterogeneity of responses to individual peptides and Ags in patients with the same haplotype points to the need to construct vaccines of multiple peptides or Ags to maximize the proportion of responding patients. PMID- 9862733 TI - Combination chemotherapy and IL-15 administration induce permanent tumor regression in a mouse lung tumor model: NK and T cell-mediated effects antagonized by B cells. AB - Previous studies have demonstrated that IL-15 administration after cyclophosphamide (CY) injection of C57BL/6J mice bearing the i.m. 76-9 rhabdomyosarcoma resulted in a significant prolongation of life. In the present study, we investigated the immune response against the 76-9 experimental lung metastases after CY + IL-15 therapy. Administration of CY + IL-15, but not IL-15 alone, induced prolongation of life and cures in 32% of mice bearing established experimental pulmonary metastases of 76-9 tumor. The CY + IL-15 therapy resulted in increased levels of NK1.1+/LGL-1+ cells, and CD8+/CD44+ T cells in PBL. In vitro cytotoxic assay of PBL indicated the induction of lymphokine-activated killer cell activity, but no evident tumor-specific class I-restricted lytic activity. Survival studies showed that the presence of NK and T lymphocytes is necessary for successful CY + IL-15 therapy. Experiments using knockout mice implied that either alphabeta or gammadelta T cells were required for an antitumor effect induced by CY + IL-15 therapy. However, mice lacking in both alphabeta and gammadelta T cells failed to respond to combination therapy. Cured B6 and alphabeta or gammadelta T cell-deficient mice were immune to rechallenge with 76-9, but not B16LM tumor. B cell-deficient mice showed a significant improvement in the survival rate both after CY and combination CY + IL-15 therapy compared with normal B6 mice. Overall, the data suggest that the interaction of NK cells with tumor-specific alphabeta or gammadelta T lymphocytes is necessary for successful therapy, while B cells appear to suppress the antitumor effects of CY + IL-15 therapy. PMID- 9862734 TI - Identification of new melanoma epitopes on melanosomal proteins recognized by tumor infiltrating T lymphocytes restricted by HLA-A1, -A2, and -A3 alleles. AB - To isolate melanoma Ags recognized by T cells, cDNA libraries made from melanoma cell lines were screened with four CTLs derived from tumor infiltrating lymphocytes (TIL) that were able to recognize melanoma cells in a HLA-A1, -A2, or -A3 restricted manner. Although cDNAs encoding the previously identified melanoma Ags, tyrosinase and gp100, were isolated, these TIL were found to recognize previously unidentified peptides. An HLA-A1-restricted CTL, TIL1388, was found to recognize a tyrosinase peptide (SSDYVIPIGTY), and an HLA-A3-restricted CTL, TIL1351, recognized a gp100 peptide (LIYRRRLMK). CTL clones isolated from the HLA A2-restricted TIL1383 recognized a gp100 peptide (RLMKQDFSV). HLA-A2-restricted CTL, TIL1200, recognized a gp100 peptide (RLPRIFCSC). Replacement of either cysteine residue with alpha-amino butyric acid in the gp100 peptide, RLPRIFCSC, enhanced CTL recognition, suggesting that the peptide epitope naturally presented on the tumor cell surface may contain reduced cysteine residues. Oxidation of these cysteines might have occurred during the course of the synthesis or pulsing of the peptide in culture. These modifications may have important implications for the development of efficient peptide-based vaccines. These newly identified peptide epitopes can extend the ability to perform immunotherapy using synthetic peptides to a broader population of patients, especially those expressing HLA-A1 or HLA-A3 for whom only a few melanoma epitopes have previously been identified. PMID- 9862735 TI - Persistent expression of experimental autoimmune encephalomyelitis (EAE)-specific Vbeta8.2 TCR spectratype in the central nervous system of rats with chronic relapsing EAE. AB - Monitoring the TCR repertoire is indispensable for the assessment of T cell associated autoimmune diseases and subsequent TCR-based immunotherapy. In the present study, we examined the TCR repertoire of spinal cord T cells of Lewis rats by CDR3 spectratyping during chronic relapsing experimental autoimmune encephalomyelitis (EAE) induced by immunization with spinal cord homogenate. It was found that Vbeta8.2 spectratype with the shortest CDR3 expanded oligoclonally throughout the course of the disease. In addition, Vbeta12 spectratype expansion was observed at the first and second attacks of EAE. Sequence analysis revealed that clones with the DSSYEQYF sequence, which is a representative sequence of myelin basic protein (MBP)-reactive T cell clones, constituted the predominant population in the Vbeta8.2 family. Surprisingly, Vbeta12 also used the identical amino acid sequence in the CDR3 region. These findings indicate that although infiltrating T cells in the central nervous system are activated polyclonally, the TCR repertoire remains unchanged throughout the course. Moreover, the finding that the predominant CDR3 amino acid sequence of Vbeta8.2 and Vbeta12 spectratypes is identical with that of MBP-induced EAE suggests that a single Ag in spinal cord homogenate, possibly MBP, is involved in disease development. PMID- 9862736 TI - Genetic determinants of autoimmune disease and coronary vasculitis in the MRL lpr/lpr mouse model of systemic lupus erythematosus. AB - MRL-lpr/lpr (MRL/lpr) mice are a model of human autoimmune disease. They exhibit a number of characteristics of systemic lupus erythematosus, including anti-DNA Abs, anti-cardiolipin Abs, immune complex-mediated vasculitis, lymphadenopathy, and severe glomerulonephritis. Although the autoimmune disorder is mediated primarily by mutation of the Fas gene (lpr), which interferes with lymphocyte apoptosis, MRL/lpr mice also have other predisposing genetic factors. In an effort to identify these additional factors, we have applied quantitative trait locus (QTL) mapping using an intercross between MRL/lpr mice and the nonautoimmune inbred strain BALB/cJ. A complete linkage map spanning the entire genome was constructed for 189 intercross progeny, and genetic loci contributing to features of the autoimmunity were identified using statistical analytic procedures. As expected, the primary genetic determinant of autoimmune disease in this cross was the Fas gene on mouse chromosome 19, exhibiting a lod score of 60. In addition, two novel loci, one on chromosome 2 (lod score, 4.3) and one on chromosome 11 (lod score, 3.1), were found to contribute to levels of anti-DNA Abs. Interestingly, the chromosome 19 and chromosome 11 QTLs, but not the chromosome 2 QTL, also exhibited associations with anti-cardiolipin Abs (lod scores, 38.4 and 2.6). We further examined the effects of these QTLs on the development of coronary vasculitis in the F2 mice. Our results indicate that the QTLs on chromosomes 11 and 19 also control the development of vasculitis, demonstrating common genetic determinants of autoantibody levels and vasculitis. PMID- 9862737 TI - Environmental antigen-induced IL-13 responses are elevated among subjects with allergic rhinitis, are independent of IL-4, and are inhibited by endogenous IFN gamma synthesis. AB - Human immediate hypersensitivity diseases represent the most common example of chronic excessive Th2-like activation in developed nations. While IL-13 shares many functional properties with IL-4, the intensity and regulation of environmental Ag-stimulated IL-13 synthesis by allergic vs nonallergic individuals remain ill defined. Here, we examine the intensity of polyclonally and Ag-stimulated IL-13 production by PBMC of 20 subjects with seasonal allergic rhinitis and 20 healthy controls. Polyclonally driven IL-13 responses did not differ significantly (Mann-Whitney U test, p = 0.68). In contrast, the median CD4 dependent IL-13 response among atopics was markedly stronger than nonatopics in Ag-stimulated primary culture (p = 0.0031) and exhibited a strong correlation with IL-5 (r = 0.76, p = 0.0009), but not IL-4 (r = 0.14, p > 0.05), responses. IL-13 production was unaffected by blocking endogenous IL-4 or IL-5 activity or by addition of rIL-4 or rIL-5. In contrast, it was inhibited by low levels of rIFN-gamma and strongly enhanced upon addition of neutralizing anti-IFN-gamma mAb. Collectively, the data are consistent with a negative regulatory role for endogenous IFN-gamma synthesis in controlling the intensity of systemic IL-13 responses evoked in both atopic and nonatopic populations following exposure to common Ags. They also suggest that the elevated levels of IL-4 and IL-5 characteristic of type 2-dominated responses in vivo are without detectable impact on the maintenance of recall Ag-stimulated IL-13 production. PMID- 9862738 TI - Prevention of experimental allergic encephalomyelitis via inhibition of IL-12 signaling and IL-12-mediated Th1 differentiation: an effect of the novel anti inflammatory drug lisofylline. AB - Experimental allergic encephalomyelitis (EAE) is an inflammatory, CD4+ Th1 mediated autoimmune disease, which serves as a model for multiple sclerosis. We examined the effect of a novel anti-inflammatory drug, lisofylline (LSF), on EAE induced either by injection of mouse spinal cord homogenate or following transfer of myelin basic protein-reactive T cells. Orally administered LSF significantly inhibited EAE in both cases, decreasing peak clinical scores by >70% and >80%, respectively. In addition, analysis of representative spinal cord sections from LSF-treated mice showed complete lack of demyelination and lymphocyte infiltration. The reduction in EAE correlated with the inhibition of Th1 differentiation by LSF in vivo, as indicated by a reduction in T cell IFN-gamma production ex vivo after Ag restimulation. The inhibition of Th1 differentiation in vivo is consistent with a block in IL-12 receptor signaling, because LSF blocked IL-12-driven Th1 differentiation and T cell proliferation in vitro, yet had no effect on IL-12 secretion from APCs ex vivo or in vitro. PMID- 9862739 TI - Terminal deoxynucleotidyl transferase deficiency reduces the incidence of autoimmune nephritis in (New Zealand Black x New Zealand White)F1 mice. AB - Terminal deoxynucleotidyl transferase (TdT) enzyme activity in lymphocytes generates diversity in the Ag receptor repertoires by adding template-independent N nucleotides and disrupting homology-directed rearrangements. The importance of this diversity in vivo and the significance of the suppression of TdT during fetal life remain uncertain. Previous studies have shown that in TdT knockout mice (TdT(0)) 1) the T cell repertoire is less peptide oriented; and 2) natural autoantibody, particularly anti-DNA autoantibodies, are less polyreactive, and their mean affinities are reduced. Consequently, the suppression of TdT during early T/B cell ontogeny may participate in controlling autoimmunity. To study the impact of TdT suppression in autoimmune-prone mice, we introduced the TdT null mutation into the (NZB x NZW)F1 (B/W) mouse strain. We show that TdT deficiency significantly reduces the incidence of autoimmune nephritis and prolongs survival compared with those in control mice. Surprisingly, the long-term survivor TdT(0) mice produced amounts of anti-ADN and anti-histone autoantibodies similar to those of their TdT+ littermates. However, these TdT(0) mice showed no evidence of renal inflammation, and the immune deposits were restricted to the mesangium, whereas basal membrane deposits were clearly correlated with overt renal disease. The present study supports the idea that the absence of TdT enzyme activity in lymphocytes protects mice against autoimmunity and could offer a therapeutic approach to autoimmune diseases. Moreover, our results may help to unravel the mechanisms of lupus nephritis. PMID- 9862740 TI - Molecular and immunologic characterization of a highly cross-reactive two EF-hand calcium-binding alder pollen allergen, Aln g 4: structural basis for calcium modulated IgE recognition. AB - Serum IgE was used to isolate a cDNA coding for a 9.4-kDa two EF-hand calcium binding allergen, Aln g 4, from a lambda gt11 expression cDNA library constructed from alder (Alnus glutinosa) pollen. rAln g 4 was overexpressed in Escherichia coli and purified to homogeneity. It reacted with serum IgE from 18% of pollen allergic patients (n = 122); shared IgE epitopes with homologous allergens present in tree, grass, and weed pollens; and thus belongs to a family of highly cross-reactive pollen allergens. Exposure of two E. coli-expressed rAln g 4 fragments comprising amino acids 1-41 and 42-85 to patients' IgE Abs, as well as to a rabbit antiserum raised against purified rAln g 4, indicated that most of the B cell epitopes reside in the N-terminal portion of the protein. IgE recognition of Aln g 4 was strongly modulated by the presence or absence of calcium. Circular dichroism analysis of rAln g 4 revealed that the protein consisted mostly of alpha helical secondary structure and possessed a remarkable thermal stability and refolding capacity, a property that was greatly reduced after calcium depletion. Circular dichroism analysis of the calcium-bound and apo form of rAln g 4 indicated that calcium-induced modulation of IgE binding could be due to changes in the protein conformation. Purified rAln g 4 elicited dose dependent basophil histamine release and immediate type skin reactions in sensitized patients. It may hence be useful for allergy diagnosis and for specific immunotherapy. PMID- 9862741 TI - Characterization of the pathogenic autoreactive T cells in cyclosporine-induced syngeneic graft-versus-host disease. AB - Administration of the immunosuppressive drug cyclosporine after syngeneic bone marrow transplantation paradoxically elicits a systemic autoimmune syndrome resembling graft-vs-host disease (GVHD). This syndrome, termed syngeneic GVHD, is associated with the development of CD8+ cytolytic T lymphocytes that promiscuously recognize MHC class II molecules in association with a peptide from the invariant chain (CLIP). Clonal analysis reveals a major subset of cells that are pathogenic and require the N-terminal flanking region of CLIP for activation, while there is a minor subset of nonpathogenic T cells that require the C terminal flanking region. The present studies show that pathogenic T cells produce type 1 cytokines (IL-2; IFN-gamma), while the nonpathogenic clones produce type 2 cytokines (IL-4; IL-10). Moreover, the repertoire of the pathogenic T cells is highly conserved with respect to V beta and V alpha TCR gene expression. The vast majority of clones express V beta8.5 (12/12) and V alpha11 (11/12). Although a limited number was evaluated, the nonpathogenic clones have only a V alpha restriction. Sequence analysis of the pathogenic T cell clones reveals a marked heterogeneity in the complementarity-determining region 3 domain and differential J region gene expression for both TCR alpha- and beta-chains. Evaluation of the specificity of these clones suggests that the functional interaction between the N-terminal flanking region of CLIP (defined by the amino acid sequence -KPVSP-) and the V region of the TCR is critical, allowing effective target cell recognition and tissue destruction in syngeneic GVHD. PMID- 9862742 TI - Temporal role of chemokines in a murine model of cockroach allergen-induced airway hyperreactivity and eosinophilia. AB - The increase in inner-city asthma among children appears to be due to allergic responses to several allergens. Recent studies have demonstrated that Ags derived from cockroaches are especially prominent in these settings and a significant health concern for the induction of asthma in children. In the present study, we have outlined the development of a murine model of cockroach allergen-induced airway disease and assessed specific mechanisms of the response, which resembles atopic human asthma. The allergic responses in this model include allergen specific airway eosinophilia and significantly altered airway physiology, which directly correlates with inflammation. We have further utilized this allergen to establish primary and secondary rechallenge stages of late phase hyperreactivity exacerbation. This latter stage is characterized by greater changes in airway physiology than the primary stage, and it is likely due to the preexisting peribronchial inflammation present at the time of the second allergen rechallenge. We have identified specific roles for CC chemokines during these stages, with MIP-1alpha being an important eosinophil attractant during the primary stage and eotaxin during the secondary rechallenge stage. The development of these models allows the evaluation of mediators involved in both stages of cockroach allergen challenge, as well as the testing of specific therapeutic modalities. PMID- 9862743 TI - Immunostimulatory DNA sequences inhibit IL-5, eosinophilic inflammation, and airway hyperresponsiveness in mice. AB - We have used a mouse model of allergen-induced airway hyperresponsiveness to demonstrate that immunostimulatory DNA sequences (ISS) containing a CpG DNA motif significantly inhibit airway eosinophilia and reduce responsiveness to inhaled methacholine. ISS not only inhibited eosinophilia of the airway (by 93%) and lung parenchyma (91%), but also significantly inhibited blood eosinophilia (86%), suggesting that ISS was exerting a significant effect on the bone marrow production of eosinophils. The inhibition of the bone marrow production of eosinophils by 58% was associated with a significant inhibition of T cell-derived cytokine generation (IL-5, granulocyte-macrophage CSF, and IL-3). ISS exerted this inhibitory effect on T cell cytokine production indirectly by stimulating monocytes/macrophages and NK cells to generate IL-12 and IFNs. The onset of the ISS effect on reducing the number of tissue eosinophils was both immediate (within 1 day of administration) and sustained (lasted 6 days), and was not due to ISS directly inducing eosinophil apoptosis. ISS was effective in inhibiting eosinophilic airway inflammation when administered either systemically (i.p.), or mucosally (i.e., intranasally or intratracheally). Interestingly, a single dose of ISS inhibited airway eosinophilia as effectively as daily injections of corticosteroids for 7 days. Moreover, while both ISS and corticosteroids inhibited IL-5 generation, only ISS was able to induce allergen-specific IFN gamma production and redirect the immune system toward a Th1 response. Thus, systemic or mucosal administration of ISS before allergen exposure could provide a novel form of active immunotherapy in allergic diseases. PMID- 9862745 TI - Dithiocarbamate pesticides affect glutamate transport in brain synaptic vesicles. AB - Dithiocarbamate compounds are widely used agricultural fungicides that display low acute toxicity in mammals and that may become neurotoxic after prolonged exposure. Mancozeb, among other dithiocarbamates tested, proved to be the most potent (Ki= 0.27 microM) at noncompetitively inhibiting the in vitro ATP dependent uptake of [3H]glutamate in rat cortical vesicles. Furthermore, mancozeb partially (20%) inhibited the ATP-dependent uptake of [14C]methylamine, used as an index for the vesicular transmembrane proton gradient (DeltapH), and evoked its efflux from organelles previously incubated with the 3H-labeled marker. Meanwhile, the vesicular uptake of 36chloride- anions whose concentrations regulate the transmembrane potential gradient (DeltapsiSV) was not impaired. The dithiocarbamate effects on the vesicular transport of [3H]glutamate thus appeared to involve mainly the DeltapH gradient rather than the potential gradient. Dithiocarbamate metabolites, the potent neurotoxin carbon disulfide included, did not affect the uptake process, thus implying the relevance for inhibition of the persistence, if any, of parent compounds in the brain. The present novel and potent in vitro interferences of selected dithiocarbamate pesticides with the vesicular transport of glutamate, if representative of in vivo alterations, may play some role in the probably complex origin of dithiocarbamate neurotoxicity. PMID- 9862744 TI - Perforin/Fas-ligand double deficiency is associated with macrophage expansion and severe pancreatitis. AB - We report that perforin/Fas-ligand double-deficient mice die early of severe pancreatitis. Female mice, in addition, are infertile and suffer from hysterosalpingitis. Tissue destruction is accompanied by infiltration with Mac-1 (CD11b)-positive monocytes/macrophages, Mac-1-positive T cells, and expansion of CD8+ T cells. In vivo inactivation of monocytes/macrophages by carrageenan reverses disease progression and restores fertility of female mice. Perforin/Fas ligand double-deficient CD4+ or CD8+ CTL are unable to lyse cognate-activated macrophages, and therefore are unable to mediate negative feedback regulation by lysis of APCs, thereby preventing further T cell activation. These studies demonstrate a novel role for perforin in homeostatic regulation of the immune response. PMID- 9862746 TI - ONO-1603, a potential antidementia drug, delays age-induced apoptosis and suppresses overexpression of glyceraldehyde-3-phosphate dehydrogenase in cultured central nervous system neurons. AB - Primary cultures of rat cerebral cortical cells and cerebellar granule cells die by an apoptotic mechanism after more than 2 weeks in cultures in the absence of medium change and glucose supplement, a process termed age-induced apoptosis of cultured neurons. Our preliminary study has shown that age-induced apoptosis of cerebellar granule cells is protected by pretreatment with tetrahydroaminoacridine (THA), an antidementia drug. In this study, we systematically compared the neuroprotective effects of THA with those of (S)-1-[N (4-chlorobenzyl)succinamoyl]pyrrolidine-2-carbaldehyde (ONO-1603), a novel prolyl endopeptidase inhibitor and potential antidementia drug. Both ONO-1603 and THA effectively delay age-induced apoptosis of cerebral and cerebellar neurons, as demonstrated morphologically with toluidine blue and fluorescein diacetate/propidium iodide staining or biochemically by DNA laddering analysis on agarose gels. ONO-1603 is about 300 times more potent than THA, with a maximal protective effect at 0.03 and 10 microM, respectively. ONO-1603 shows a wide protective range of 0.03 to 1 microM in contrast to a narrow effective range of 3 to 10 microM for THA. Moreover, ONO-1603 is nontoxic to neurons, even at the high concentration of 100 microM, whereas THA elicits severe neurotoxicity at a dose of >/=30 microM. Both ONO-1603 and THA robustly suppress overexpression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) mRNA and accumulation of GAPDH protein in a particulate fraction of cultured neurons undergoing age-induced apoptosis. Because we documented that GAPDH overexpression participates in neuronal apoptosis induced by various insults, we conclude that the neuroprotective actions of ONO-1603 and THA appear to be mediated by suppression of this protein overexpression. PMID- 9862747 TI - Blockade of striatal dopamine transporters by intravenous methylphenidate is not sufficient to induce self-reports of "high". AB - The reinforcing effects of cocaine and methylphenidate have been linked to their ability to block dopamine transporters (DAT). Using positron emission tomography (PET), we previously showed that intravenous cocaine induced a significant level of DAT blockade, which was associated with the intensity for self-reports of "high" in cocaine abusers. In this study, we measured DAT occupancies after intravenous methylphenidate and assessed whether they also were associated with the "high". Occupation of DAT by intravenous MP was measured with PET using [11C]cocaine, as a DAT ligand, in eight normal control subjects tested with different methylphenidate doses. The ratio of the distribution volume of [11C]cocaine in striatum to that in cerebellum, which corresponds to Bmax/Kd + 1, was used as measure of DAT availability. In parallel, self-reports of "high" were measured. Methylphenidate produced a dose-dependent blockade of DAT with an estimated ED50 of 0.075 mg/kg. DAT occupancies were significantly correlated with the "high" (p <.03). However, four of the eight subjects, despite having significant levels of DAT blockade, did not perceive the "high". Methylphenidate is as effective as cocaine in blocking DAT in the human brain (cocaine ED50 = 0.13 mg/kg), and DAT blockade, as for cocaine, was also associated with the "high". However, the fact that there were subjects who despite significant DAT blockade did not experience the "high" suggests that DAT blockade, although necessary, is not sufficient to produce the "high". PMID- 9862748 TI - Three-dimensional quantitative structure activity relationship analyses of substrates for CYP2B6. AB - To begin to build an understanding of the interactions of CYP2B6 with substrates, two different 3-dimensional quantitative structure activity relationship (3D QSAR) models were constructed using 16 substrates of B-lymphoblastoid expressed CYP2B6. A pharmacophore model was built using the program Catalyst, which was compared with a partial least-squares (PLS) model using molecular surface weighted holistic invariant molecular (MS-WHIM) descriptors. The Catalyst model yielded a 3-dimensional model of the common structural features of CYP2B6 substrates, whereas PLS MS-WHIM generated a model based on statistical analyses of molecular descriptors for size and shape of the substrate. The pharmacophore model obtained with Catalyst consisted of three hydrophobes and one hydrogen bond acceptor region. The cross-validated PLS MS-WHIM model gave a good q2 value of 0.607. Size, positive electrostatic potential, hydrogen bonding acceptor capacity, and hydrophobicity were found to be the most relevant descriptors for the model. These models were then used to predict the Km (apparent) values of a test set of structurally diverse substrates for CYP2B6 not included in the model building, specifically lidocaine, amitriptyline, bupropion, arteether, and verapamil. Overall, both 3D-QSAR methods yielded satisfactory Km (apparent) value predictions for the majority of the molecules in the test set. However, PLS MS WHIM was unable to reliably predict the Km (apparent) value for verapamil, whereas Catalyst did not predict the Km (apparent) value for lidocaine. In both of these cases the residual of the Km (apparent) value was greater than one log unit. The strengths and limitations of both of these 3D-QSAR approaches are discussed. PMID- 9862749 TI - Chronic morphine treatment selectively augments metabotropic glutamate receptor induced inhibition of N-methyl-D-aspartate receptor-mediated neurotransmission in nucleus accumbens. AB - We compared the effects of different metabotropic glutamate receptor (mGluR) agonists on pharmacologically isolated N-methyl-D-aspartate-excitatory postsynaptic currents (NMDA-EPSCs) in core nucleus accumbens neurons using conventional intracellular recording in untreated and morphine-treated rats. The rats were treated by s.c. implantation of two morphine pellets and studied over a 3- to 6-day period. This model is known to exhibit opiate tolerance and dependence. We elicited NMDA-EPSCs by stimulating locally in the presence of the alpha-amino-3-hydroxy-5-methly-4-isoxazolepropionic acid/kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (10 microM) and the gamma aminobutyric acid receptor antagonist bicuculline (15 microM). We found that trans-1-aminocyclopentane-1,3-decarboxylic acid, an agonist of group 1 and 2 mGluRs, decreased NMDA-EPSC areas (time-integrals) in a dose-dependent manner (1 10 microM) in slices taken from untreated rats. This inhibitory effect was significantly enhanced after chronic morphine treatment. In contrast, although the group 3 mGluR agonist L(+)-2-amino-4-phosphonobutyric acid also markedly reduced NMDA-EPSC areas, there was no apparent change in this effect after chronic morphine. We found that quisqualate, the group 1 mGluR agonist, failed to elicit any effect on NMDA-EPSCs in either untreated or chronically treated rats. Paired-pulse stimulation of core nucleus accumbens NMDA-EPSCs in slices from these groups showed that chronic morphine enhanced paired-pulse facilitation, consistent with a presynaptic reduction in glutamate release. Because of the relevance to opiate tolerance and dependence of the chronic model used, the brain region (accumbens), and the receptors studied, our data provide a cellular substrate that could account for some aspects of these phenomena. PMID- 9862750 TI - RhoA-sensitive trafficking of muscarinic acetylcholine receptors. AB - The clathrin-mediated sequestration pathway is used by non-G protein-coupled receptors (e.g., transferrin receptors) and a large number of G protein-coupled receptors, including beta-2 adrenoceptors and various muscarinic acetylcholine receptor (mAChR) subtypes. Recently, the ubiquitously expressed small GTPase RhoA has been implicated as a negative regulator of transferrin receptor internalization. Because mAChRs and other G protein-coupled receptors are able to activate RhoA, we investigated in HEK-293 cells whether RhoA regulates the sequestration of m1 and m2 mAChRs, which internalize via clathrin-coated and nonclathrin-coated vesicles in HEK-293 cells, respectively. Overexpression of wild-type RhoA inhibited agonist-induced sequestration of both m1 and m2 mAChRs by as much as 70%. Inhibition could be reversed by coexpression of Clostridium botulinum C3 transferase, which inactivates RhoA by ADP-ribosylation. Overexpression of C3 transferase alone had no effect on m1 and m2 mAChR sequestration. In addition, overexpression of RhoA inhibited m1 and m2 mAChR transport to the plasma membrane by 60 and 31%, respectively, which was blocked by coexpression of C3 transferase. We conclude that RhoA is not an endogenous regulator of mAChR sequestration, but when overexpressed, strongly inhibits mAChR trafficking (i.e., sequestration and transport to the plasma membrane) in HEK-293 cells. PMID- 9862752 TI - Detection of 2-hydroxyiminostilbene in the urine of patients taking carbamazepine and its oxidation to a reactive iminoquinone intermediate. AB - Carbamazepine is one of the most widely used anticonvulsants in North America; however, its use is associated with a range of serious idiosyncratic adverse reactions. These reactions are thought to result from the formation of chemically reactive metabolites. Carbamazepine is extensively metabolized in the liver and one of the major metabolites is 2-hydroxycarbamazepine, which has previously been detected as a urinary metabolite excreted by rats and humans along with its further metabolized product, 2-hydroxyiminostilbene. In this study, we found that the urine of patients taking carbamazepine appeared to contain more of the glucuronide of 2-hydroxyiminostilbene than that of 2-hydroxycarbamazepine. We have also demonstrated that 2-hydroxyiminostilbene can be oxidized readily to an iminoquinone species by HOCl, H2O2 or even on exposure to air. The reactivity of this iminoquinone as an electrophile was studied. It was shown to react with sulfhydryl-containing nucleophiles, such as glutathione and N-acetylcysteine. We also found a metabolite with the same molecular weight as 4-methylthio-2 hydroxyiminostilbene, but not the corresponding carbamazepine derivative, in the urine of patients taking carbamazepine and this presumably reflects the formation of a glutathione conjugate of the reactive iminoquinone. This iminoquinone intermediate may play a role in carbamazepine-induced idiosyncratic reactions. PMID- 9862751 TI - Effect of overproduction of interleukin 5 on dinitrofluorobenzene-induced allergic cutaneous response in mice. AB - The effect of overproduction of interleukin (IL) 5 on the allergic cutaneous response was investigated in transgenic mice overexpressing IL-5. Five repeated topical applications of 2, 4-dinitrofluorobenzene (DNFB) to the ears of mice resulted in allergic dermatitis on the ears as well as significant elevation in dinitrophenol-specific IgE antibody and total IgE in the serum in both wild-type and transgenic mice. The development of dermatitis as measured by skin thickness and histopathological changes were potentiated in the transgenic mice. In IL-5 transgenic mice, significant accumulation of eosinophils in skin lesions was observed after five paintings of DNFB, and the magnitudes of eosinophilia and IL 5 messenger RNA expression were significantly higher than in wild-type mice. The dinitrophenol-specific and total IgE in the serum were higher in IL-5 transgenic mice. The late phase reaction of IgE antibody-mediated biphasic cutaneous response was potentiated in IL-5 transgenic mice. The magnitudes of vasopermeability increase by passive cutaneous anaphylaxis, serotonin, and platelet-activating factor were similar in both mice. These results indicate that overproduction of IL-5 resulted in the potentiation of DNFB-induced dermatitis by elevation of IgE production, IgE-mediated allergic late-phase cutaneous reaction, and eosinophilia in the skin lesion. PMID- 9862753 TI - Renal drug targeting using a vector "alkylglycoside". AB - A specific sugar-modified peptide has previously been shown to have renal targeting potential in vivo and to have a specific binding site which has been identified in the kidney membrane fraction. In this report, we studied the inhibitory effects of glycosylated derivatives on the binding of [3H]Glc-O-C8-AVP [a glucosylated derivative of Arg8-vasopressin (AVP), Kd = 55 nM] to clarify the structural requirements necessary for renal recognition. Glc-S-C7-Me (octyl beta D-thioglucoside) markedly inhibited the binding, to a much greater extent than Glc-O-C7-Me (octyl beta-D-glucoside) and Gal-S-C7-Me (octyl beta-D thiogalactoside). Also, [3H]Glc-S-C7-Me was shown to have a specific binding site on the kidney membrane (Kd = 17 nM, Bmax = 24 pmol/mg protein) rather than the liver membrane and, in addition, Glc-S-C7-Me exhibited effective and selective renal uptake in vivo. To examine the possibility that Glc-S-C7-Me might be of practical use as a renal targeting vector, AVP, tryptamine and 4-nitrobenz-2-oxa 1,3-diazole were modified with Glc-S-C8- and the tissue uptake of the resulting derivatives was evaluated. All of these derivatives showed clear renal targeting potential because the apparent uptake clearance by the kidney was greater than 3 ml/min/g kidney in each case. As far as the AVP derivatives were concerned, derivatives having different numbers of methylene groups were compared with Glc-S C8-AVP. Glc-S-C11-AVP exhibited increased kidney targeting potential, whereas that of Glc-S-C5-AVP was reduced. These differences suggest that the "alkylglycoside" moiety is important for renal uptake. In addition, these renally targeted derivatives inhibited the binding of [3H]Glc-S-C7-Me to the kidney membrane fraction. Our findings allow us to conclude that the alkylglycoside is a suitable candidate vector for renal targeting. PMID- 9862754 TI - Diclofenac toxicity to hepatocytes: a role for drug metabolism in cell toxicity. AB - Diclofenac, a 2-arylacetic acid, nonsteroidal anti-inflammatory drug, has been reported to cause adverse hepatic effects in certain individuals. To discriminate among possible mechanisms of hepatotoxicity, we examined the effects of diclofenac on human and rat hepatocytes and hepatic cell lines (HepG2, FaO), investigated the major biochemical events in the course of diclofenac cytotoxicity (calcium homeostasis, lipid peroxidation, and mitochondrial dysfunction), and investigated whether cytotoxicity could be related to drug metabolism by cytochrome P-450. Acute diclofenac-induced toxicity in hepatocytes was preluded by a decrease in ATP levels, whereas no significant oxidative stress (decrease in glutathione and lipid peroxidation) or increase in intracellular calcium concentration could be observed at early incubation stages. Diclofenac was more cytotoxic to drug metabolizing cells (rat and human primary cultured hepatocytes) than to nonmetabolizing cell lines (HepG2, FaO). Despite the fact that diclofenac itself was effective in impairing ATP synthesis by mitochondria, we found evidence that toxicity was also related to drug metabolism and was reduced by the addition of cytochrome P-450 inhibitors (proadifen and ketoconazole) to culture medium. The in vitro cytotoxicity correlated well with the formation by hepatocytes of 5-hydroxydiclofenac and, in particular, N,5 dihydroxydiclofenac, a minor metabolite first characterized in this article. Hepatic microsomes showed the ability to both oxidize 5-hydroxydiclofenac to N,5 dihydroxydiclofenac and back reduce the latter to 5-hydroxydiclofenac with the consumption of NADPH. The experimental results suggest that the toxic effect of diclofenac on hepatocytes may be caused by drug-induced mitochondrial impairment, together with a futile consumption of NADPH. PMID- 9862755 TI - Neomycin inhibits catecholamine secretion by blocking nicotinic acetylcholine receptors in bovine adrenal chromaffin cells. AB - We investigated the effects of neomycin on nicotinic acetylcholine receptor induced responses in bovine adrenal chromaffin cells. Neomycin inhibited the nicotinic agonist dimethylphenylpiperazinium iodide (DMPP)-induced norepinephrine secretion in a concentration-dependent manner. Neomycin had also an inhibitory effect on the DMPP-induced increase in cytosolic Ca++ concentration ([Ca++]i). This effect was further confirmed by inhibition of the DMPP-induced fluorescence quenching of fura-2 upon Mn++ entry. Under the same conditions, however, neomycin did not change the bradykinin-induced [Ca++]i increase, which follows the downstream signal of phospholipase C phospholipase C activation in this cell. The inhibitory effect of neomycin on the DMPP-induced [Ca++]i increase was apparent when the neomycin treatment was performed simultaneously with DMPP, suggesting a direct action on the nicotinic receptor. The direct inhibitory action of neomycin on the nicotinic receptor was also evident when neomycin inhibited the DMPP induced cytosolic Ca++ increase, which is not affected by nifedipine nor omega conotoxin MVIIC, and the cytosolic Na+ increase, which is not affected by tetrodotoxin. In addition, we observed that neomycin inhibited the binding of nicotine to the acetylcholine receptor in a noncompetitive manner. The data suggest that neomycin inhibits the nicotinic acetylcholine receptor directly, which results in blockage of the nicotinic receptor-mediated signaling without involvement of phospholipase C. PMID- 9862756 TI - Chronic clomipramine and triiodothyronine increase serotonin levels in rat frontal cortex in vivo: relationship to serotonin autoreceptor activity. AB - Augmentation of tricyclic antidepressant (TCA) treatment with triiodothyronine (T3) has been shown to potentiate the therapeutic effect of TCA drugs in depressed patients. We have attempted to elucidate the mechanism of this potentiation by determining the effects of T3 alone and together with a TCA on serotonin (5-HT) levels in living rats, using in vivo microdialysis. A single s.c. injection of T3 at 0.1 mg/kg had no effect on 5-HT levels in frontal cortex or hippocampus. Chronic administration of clomipramine (10 mg/kg i.p. daily for 4 weeks) to rats resulted in increased basal 5-HT levels in the frontal cortex. Administration of T3 daily for 7 days at 0.1 mg/kg s.c. also resulted in elevated 5-HT levels, whereas in rats administered both clomipramine and T3, cortical 5-HT levels were significantly elevated compared with the levels in rats that had received only one treatment. Basal levels in hippocampus were unaffected by these treatments. Subcutaneous injection of the 5-HT-1a receptor agonist 8-hydroxy-2 (di-n-propylamino)tetralin (0.2 mg/kg) resulted in a decrease in 5-HT levels in both cortex and hippocampus. In frontal cortex of animals that had received T3 or a combination of clomipramine and T3, the extent of the decrease was significantly reduced compared to that seen in control animals. The extent of the decrease in hippocampus was not affected by any of the treatments. Subcutaneous injection of the 5-HT-1b/1d antagonist GR 127935 (5 mg/kg) resulted in an increase in 5-HT levels in both brain areas. The extent of the increase was not affected by any of the treatments in either brain area. It is concluded that the action of T3 in potentiating the clinical response to TCA drugs may be due to its effect on 5-HT levels in the frontal cortex, which is due to desensitization of the presynaptic 5-HT-1a autoreceptors. PMID- 9862757 TI - Noncompetitive functional inhibition at diverse, human nicotinic acetylcholine receptor subtypes by bupropion, phencyclidine, and ibogaine. AB - Nicotinic acetylcholine receptors (nAChR) are diverse members of the neurotransmitter-gated ion channel superfamily and play critical roles in chemical signaling throughout the nervous system. The present study establishes the acute functional effects of bupropion, phencyclidine, and ibogaine on two human nAChR subtypes. Function of muscle-type nAChR (alpha1 beta gamma delta) in TE671/RD cells or of ganglionic nAChR (alpha3 beta4 alpha5+/-beta2) in SH-SY5Y neuroblastoma cells was measured with 86Rb+ efflux assays. Functional blockade of human muscle-type and ganglionic nAChR is produced by each of the drugs in the low to intermediate micromolar range. Functional blockade is insurmountable by increasing agonist concentrations in TE671/RD and SH-SY5Y cells for each of these drugs, suggesting noncompetitive inhibition of nAChR function. Based on these findings, we hypothesize that nAChR are targets of diverse substances of abuse and agents used in antiaddiction/smoking cessation strategies. We also hypothesize that nAChR play heretofore underappreciated roles in depression and as targets for clinically useful antidepressants. PMID- 9862758 TI - Propulsion in guinea pig colon induced by 5-hydroxytryptamine (HT) via 5-HT4 and 5-HT3 receptors. AB - Previous studies have shown that the intestinal peristaltic reflex initiated by mucosal stimulation is mediated by release of 5-hydroxytryptamine (HT) from enterochromaffin cells; 5-HT acts via 5-HT4 receptors in rat and human, and via both 5-HT4 and 5-HT3 receptors in guinea pig to activate intramural sensory neurons that release calcitonin gene-related peptide. In this study, selective agonists and antagonists were used to examine the involvement of 5-HT4 and 5-HT3 receptors in colonic propulsion. The velocity of propulsion was measured with artificial fecal pellets introduced into the orad end of an isolated guinea pig colonic segment. Control velocity ranged from 0.5 to 3.3 mm/s; mean +/- S.E.M., 1.3 +/- 0.1 mm/s. The 5-HT4 antagonist, GR 113808A, and the 5-HT3 antagonist, LY 278584, decreased the velocity of pellet propulsion in a concentration-dependent fashion (39 +/- 2% and 47 +/- 1% decrease at 10 microM, respectively). A combination of both antagonists (10 microM each) was additive, decreasing the velocity by 82 +/- 3% to 84 +/- 4%. The selective 5-HT4 agonists, HTF 919 and R093877, as well as 5-HT in the presence of the 5-HT2a antagonist, ketanserin, increased the velocity of propulsion in a concentration-dependent fashion with EC50s of 6.9 +/- 0.1 nM, 37.4 +/- 1.0 nM, and 3.9 +/- 0. 1 nM, respectively. Compared with HTF 919, R093877 was less potent and appeared to be a partial agonist. All three agonists were effective at submicromolar concentrations; at concentrations above 1 microM, there was no increase in the velocity of propulsion. We conclude that the presence of fecal pellets triggers the release of 5-HT, which acts via both 5-HT3 and 5-HT4 receptors to regulate propulsive activity in guinea pig colon. PMID- 9862759 TI - Daily injections of fluoxetine induce dose-dependent desensitization of hypothalamic 5-HT1A receptors: reductions in neuroendocrine responses to 8-OH DPAT and in levels of Gz and Gi proteins. AB - The present studies examined the dose-response relationship of fluoxetine-induced desensitization of hypothalamic postsynaptic 5-HT1A receptors, as measured from the reduced neuroendocrine responses to a 5-HT1A agonist. Because hypothalamic Gz proteins mediate the ACTH and oxytocin responses to 5-HT1A receptor activation, we also determined the effect of fluoxetine on the levels of Gz proteins in the hypothalamus. Rats were injected daily for 14 days with saline or with fluoxetine doses of 0.3, 1, 3, 5, 7. 5, or 10 mg/kg/day. Fluoxetine produced a dose dependent reduction in the oxytocin, ACTH, and corticosterone responses to the 5 HT1A agonist 8-hydroxy-2-(dipropylamino)tetralin (8-OH-DPAT, 50 micrograms/kg, s.c.). The lowest fluoxetine dose that significantly, although incompletely, reduced the neuroendocrine responses to 8-OH-DPAT was 5 mg/kg/day. The 10 mg/kg/day dose of fluoxetine maximally inhibited all neuroendocrine responses to 8-OH-DPAT. Hypothalamic levels of Gz protein were reduced by both the 7.5 and 10 mg/kg/day doses of fluoxetine, whereas Gi1 protein levels were reduced only after the highest dose (10 mg/kg/day) of fluoxetine. Gi2, Gi3, and Go levels were not reduced by any fluoxetine dose. Cytosolic levels of Gi1 and Gz proteins were unaltered, indicating that reductions in Gz and Gi1 proteins are not caused by a redistribution of the proteins from the membrane into the cytosol. The results from the present study indicate that fluoxetine-induced desensitization of hypothalamic postsynaptic 5-HT1A receptor systems is dose-dependent and may be caused in part by reductions in the hypothalamic levels of Gz proteins. PMID- 9862760 TI - Increasing doses of pentoxifylline as a continuous infusion in canine septic shock. AB - We investigated effects of pentoxifylline during septic shock. Two-year-old (10 12 kg), purpose-bred beagles were infected i.p. with Escherichia coli 0111:B4 (1.2-1.5 x 10(9) colony-forming units per kilogram b.wt.) in a fibrin clot and then immediately treated with one of five doses of pentoxifylline (0.5-20 mg. kg 1. h-1 i.v.) as a 36-h continuous infusion or placebo. All animals received antibiotics and fluid resuscitation. Pentoxifylline levels increased in a dose dependent manner during (p =.001) and were undetectable 12 h after stopping the infusion. During infusion of pentoxifylline at all doses, there were increases (p =.003), and once the infusion was stopped, there were decreases (p =.049) in endotoxin levels compared with controls. After clot implantation, at all pentoxifylline doses there was a significant increase in tumor necrosis factor levels, compared with controls (p =.025). The relative risk of death was significantly increased with pentoxifylline therapy in a dose-dependent fashion (20 >/= 10 >/= 5.0 >/= 1.0 >/= 0.5 mg. kg-1, p =.008). One hypothesis consistent with these data is that high pentoxifylline levels slowed endotoxin clearance, resulting in high levels of endotoxemia and increased proinflammatory mediator release and death. Pentoxifylline, used as a long-term continuous infusion as is commonly done clinically, can be harmful during Gram-negative septic shock. PMID- 9862761 TI - Treatment with liposome-bound recombinant human tumor necrosis factor-alpha suppresses parasitemia and protects against Plasmodium berghei k173-induced experimental cerebral malaria in mice. AB - Our study describes liposomes (conventional or sterically stabilized) as carrier systems for recombinant human tumor necrosis factor-alpha (rhTNF-alpha) to increase its protective efficacy against Plasmodium berghei-induced experimental cerebral malaria (ECM) in mice. rhTNF-alpha was either covalently coupled to the outer surface of preformed liposomes or encapsulated into the liposomes. For coupling to the liposomes, reactive thiol groups were introduced in rhTNF-alpha by reaction with N-succinimidyl S-acetylthioacetate. Intravenous injection of liposome-bound rhTNF-alpha substantially enhanced protection against ECM as compared with injection of free rhTNF-alpha. A similar protective efficacy against ECM was obtained by treatment with rhTNF-alpha coupled to either conventional or sterically stabilized liposomes. Encapsulation of rhTNF-alpha into liposomes did not improve the protective efficacy of rhTNF-alpha against P. berghei-induced ECM. Parasitemia was suppressed by treatment with either free or liposome-bound rhTNF-alpha in mice protected against ECM, but not in rhTNF-alpha treated mice developing ECM. These data suggest that the effect of rhTNF-alpha on parasitemia plays a role in establishing protection against ECM. Our studies indicate that liposome-bound rhTNF-alpha exhibits an enhanced protective efficacy against ECM compared with free rhTNF-alpha. It is hypothesized that thiolation of rhTNF-alpha and coupling to the liposomal bilayer stabilizes the bioactive trimeric configuration of rhTNF-alpha. PMID- 9862762 TI - [S]-AR-R 15896AR-A novel anticonvulsant: acute safety, pharmacokinetic and pharmacodynamic properties. AB - A rational, chemical, synthetic effort to identify promising low-affinity uncompetitive N-methyl-D-aspartic acid receptor antagonists for use as antiepileptic drugs led to the discovery of AR-R 15035AR, or [RS]-alpha-phenyl-2 pyridine-ethanamine.2HCl. Chiral separation followed by intensive in vivo screening resulted in the selection of the [S] enantiomer, AR-R 15896AR, as the best compound for further preclinical development. AR-R 15896AR prevented tonic seizures in rodents for up to 6 to 8 h in response to maximal electroshock (MES), 4-aminopyridine, bicuculline, or strychnine, as well as characteristic seizures following injections of N-methyl-DL-aspartic or kainic acids. AR-R 15896AR was ineffective in two kindling models of epilepsy, did not produce tolerance to MES, and was devoid of proconvulsant and phencyclidine-like properties in mice and rats, respectively. Therapeutic indices for AR-R 15896AR were comparable to or exceeded those for standard anticonvulsants. Orally administered AR-R 15896AR rapidly entered the rat brain and was eliminated in parallel from the plasma and plasma-free compartment. A dose-response relationship between plasma and brain levels after p.o. or i.v. administration of AR-R 15896AR and protection against MES was highly correlative. The time course for loss of protection against MES mirrored the elimination of the compound from brain and plasma. The total brain concentration (25 microM) of drug at the ED50 value (approximately 3 mg/kg) for protection against MES seizures was consistent with the reported affinity of AR-R 15896AR at the N-methyl-D- aspartic acid binding site (IC50 value = 1.3 microM). The present findings demonstrated the attractiveness of AR-R 15896AR as a candidate for further development to treat epilepsy. PMID- 9862763 TI - Inhibitory effect of rebamipide on the neutrophil adherence stimulated by conditioned media from Helicobacter pylori-infected gastric epithelial cells. AB - We investigated the mechanism or mechanisms by which rebamipide protects against the gastric mucosal inflammation associated with Helicobacter pylori. The production of interleukin (IL)-8 in association with expression of IL-8 mRNA was greatly increased in the H. pylori-infected Kato III cells in a concentration- and time-dependent manner, whereas the secretion of IL-6 and tumor necrosis factor-alpha was not detectable. The increased production of IL-8 and expression of IL-8 mRNA were significantly inhibited by rebamipide (100-1000 microM) in a concentration-dependent manner. Formyl-methionyl-leucyl-phenylalanine (1 nM), as well as conditioned medium (CM) that was produced from H. pylori-infected Kato III cells, caused an increase in surface expression of CD11b on human neutrophils and an increase in neutrophil adhesion to the human umbilical vein endothelial cells. Rebamipide also suppressed the adherence of neutrophils to endothelial cells as well as the expression of CD11b on neutrophils induced by formyl methionyl-leucyl-phenylalanine and CM. Furthermore, CM-induced neutrophil adhesion to the endothelial cells was significantly inhibited by IL-8 neutralizing antibody, suggesting that IL-8 is implicated in the CM-induced neutrophil adhesion to the cultured human umbilical vein endothelial cells. It is concluded that rebamipide exerts its preventive effect against H. pylori-evoked gastric mucosal cell inflammation by inhibition of the neutrophil adherence to the endothelial cells as well as by suppressing the surface expression of CD11b on neutrophils and the production of proinflammatory cytokine such as IL-8 from gastric epithelial cells. PMID- 9862764 TI - Mechanisms of antihyperglycemic effects of moxonidine in the obese spontaneously hypertensive Koletsky rat (SHROB). AB - Increased activity of the sympathetic nervous system may be a critical factor in the development of impaired insulin secretion and insulin resistance. We studied the chronic effects of sympathetic inhibition with moxonidine on glucose metabolism in the spontaneously hypertensive genetically obese rat (SHROB). This unique animal model closely resembles human syndrome X, expressing insulin resistance, genetic obesity, spontaneous hypertension, and hyperlipoproteinemia. Moxonidine, a selective imidazoline receptor agonist, was administered to lean spontaneous hypertensive rats (SHR) and SHROBs for 90 days in food at 8 mg/kg/day and significantly reduced mean blood pressure. Moxonidine treatment reduced fasting insulin levels by 71% in SHROB and lowered plasma free fatty acids by 25%. In SHR, moxonidine treatment decreased free fatty acids by 17% compared with controls. During an oral glucose tolerance test, blood glucose levels in moxonidine-treated SHROB were reduced relative to untreated controls from 60 min onwards. Insulin secretion was facilitated at 30 min (83% greater) and 60 min (67% greater) postchallenge compared with control SHROB. In skeletal muscle, moxonidine treatment increased the expression of the insulin receptor beta subunit by 19% in SHROB but was without effect in SHR. The level of insulin receptor substrate-1 (IRS-1) protein was decreased by 60% in control SHROB compared with lean SHR. Moxonidine treatment enhanced the expression and insulin stimulated phosphorylation of IRS-1 protein in skeletal muscle in SHROB by 74 and 27%, respectively, and in SHR by 40 and 56%, respectively. Moxonidine increased the levels of expression of IRS-1 protein in liver in SHR by 275% and in SHROB by 260%. These findings indicate that chronic inhibition of sympathetic activity with moxonidine therapy can lower free fatty acids and significantly improve insulin secretion, glucose disposal, and expression of key insulin signaling intermediates in an animal model of obese hypertension. PMID- 9862766 TI - Uptake of fentanyl in pulmonary endothelium. AB - Fentanyl is a basic amine shown to have extensive first-pass pulmonary uptake. To evaluate the role of the pulmonary endothelium in this uptake process, the simultaneous pharmacokinetics of [3H]fentanyl and two marker drugs, blue dextran, and [14C]antipyrine, were evaluated in a flow-through system of pulmonary endothelial cells. Fentanyl equilibrium kinetics were determined in a static culture system. The flow-through system consisted of monolayers of bovine pulmonary artery endothelial cells cultured on solid microcarrier beads placed in a chromatography column and perfused at 1.0 ml/min (37 degreesC). Fentanyl and the markers were injected into the perfusate at the top of the column and samples were collected from the eluate at 9-s intervals for 10 min. The pharmacokinetic analyses were based on determinations of mean transit time and flow. Fentanyl was partitioned into the pulmonary endothelial cells 60 times more than the tissue water space marker antipyrine. In the static system, monolayers of bovine pulmonary artery endothelial cells were cultured in 3.8-cm2 wells to which were added 0 to 946 micromol (0-500 microgram/ml) of unlabeled fentanyl citrate and 0.14 micromol of [3H]fentanyl. After a 10-min incubation, solubilized cells were assayed for [3H]fentanyl. Pulmonary endothelial cells contained a higher relative fentanyl concentration at lower fentanyl supernatant concentrations than would be expected if uptake occurred by diffusion alone. These observations can be explained with a model of fentanyl uptake that includes both passive diffusion and saturable active uptake. This suggests that the extensive first-pass pulmonary uptake of fentanyl observed in vivo is due largely to vascular endothelial drug uptake by both a passive and a saturable active uptake process. PMID- 9862765 TI - Insights into the unusual alpha adrenoceptor subtype in dog saphenous vein using phenoxybenzamine. AB - In the dog saphenous vein (DSV), phenylephrine (PE) responses through alpha-1 adrenoceptors receptors are antagonized by both alpha-1 and alpha-2 receptor antagonists. Furthermore, pretreatment with chloroethylclonidine (CEC) eliminates prazosin binding but reduces rauwolscine binding by half (). In new functional experiments, the effects of preincubation with phenoxybenzamine (PBZ), an irreversible alpha adrenoceptor antagonist, on responses to PE and two selective alpha-2 adrenoceptor agonists were evaluated. Also, the ability of prazosin or rauwolscine to prevent irreversible losses of responses to these agonists when coincubated with PBZ was determined. Preincubation in PBZ (10-300 nM) concentration dependently reduced PE Emax and the calculated fraction of residual receptors (q). Preincubation in PBZ (10-300 nM) increased KB values for prazosin (30 and 100 nM) but did not alter the KB value for rauwolscine (50 nM) acting at the residual receptors from control values. Coincubation of PBZ with prazosin partially prevented these PBZ actions (Emax partly restored) on responses to PE, but coincubation of rauwolscine (/=300 nM caused >50% reduction in Emax values of responses but did not alter the EC50 values for either agonist. Coincubation of rauwolscine with PBZ protected responses to alpha-2 agonists against PBZ (1 microM) effects. This study shows that PE initiates contractions at atypical alpha-1 adrenoceptors represented by all sites of PE action. Rauwolscine antagonizes PE actions but does not protect against PBZ inactivation. Typical alpha-2 adrenoceptors are distinguished from the unusual alpha-1 adrenoceptors by their lesser sensitivity to PBZ and their protection by rauwolscine from PBZ. PMID- 9862767 TI - Selectivity profile of muscarinic toxin 3 in functional assays of cloned and native receptors. AB - By using acetylcholine-induced stimulation of [35S]guanosine-5'-O-(3 thio)triphosphate ([35S]GTPgammaS) binding to membrane G proteins as a functional assay of the cloned human m1-m4 muscarinic receptor subtypes stably expressed in Chinese hamster ovary cells, muscarinic toxin 3 (MT3) was found to block the m4 receptor with a potency (pA2 = 8.33) much higher than those displayed at the m1 (pA2 = 6.78), m3 (pA2 = 6.3), and m2 (pA2 < 6.3) subtypes. In N1E-115 cells, which have been reported to express m4 receptors coupled to inhibition of cAMP, MT3 potently antagonized the carbachol-induced inhibition of adenylyl cyclase with a pA2 of 8. 81 and displayed monophasic inhibitory curves. Unexpectedly, in NG108-15 cells, known to express only m4 receptors, MT3 counteracted the carbachol inhibition of adenylyl cyclase with a lower potency (pA2 = 7.60) and showed a biphasic inhibitory curve, suggesting the participation of both m4 and m2 receptors. This possibility was supported by radioligand binding data showing that MT3 failed to completely displace the binding of [3H]N-methylscopolamine to NG108-15 cell membranes and by reverse transcription-polymerase chain reaction analysis, revealing the presence of mRNAs for both m4 and m2 receptor subtypes. These data demonstrate that MT3 possesses a high functional receptor selectivity for both the cloned and native m4 receptors and that in cell systems containing m4 and m2 receptors coupled to a common response, the toxin constitutes a powerful tool to resolve the relative contribution by each receptor subtype. PMID- 9862768 TI - Modulation of the permeability of H2 receptor antagonists cimetidine and ranitidine by P-glycoprotein in rat intestine and the human colonic cell line Caco-2. AB - The influence of secretory transporters on intestinal permeability characteristics of the H2 receptor antagonists ranitidine and cimetidine was studied in Caco-2 monolayers and rat intestinal mucosa mounted in Ussing chambers. Both drugs exhibited vectorial transport across rat ileum with significantly greater (2-4-fold) permeability in the serosal-to-mucosal than the mucosal-to-serosal direction, indicative of net mucosal secretion. Mucosal ranitidine secretion was also observed in rat distal colon, although to a lesser degree. Ileal ranitidine secretion was concentration dependent and significantly reduced by the P-glycoprotein (P-gp) substrates verapamil and cyclosporin. In contrast, probenicid, an inhibitor of the multidrug-related protein, had no effect on ranitidine permeability. The paracellular marker mannitol showed no evidence of asymmetric permeability or sensitivity to P-gp inhibitors. Significant expression of P-gp protein in rat intestinal epithelial cells was confirmed by immunoblotting. Caco-2 monolayers, which overexpress P-gp, also showed asymmetric permeability of ranitidine and cimetidine. In this model, ranitidine permeability in the mucosal-to-serosal direction decreased by approximately 95% as monolayer resistance increased from 150 to 500 Omega/cm2, indicating a primarily paracellular route of transport. However, serosal-to mucosal permeability was insensitive to resistance changes, consistent with a primarily transcellular route in this direction. These data indicate that ranitidine and cimetidine can act as substrates for intestinal P-gp and suggest that the balance between absorptive and secretory mechanisms as a factor in determining intestinal absorption needs to be a routine consideration even for compounds expected to have a predominantly paracellular route of absorption. PMID- 9862769 TI - Altered plasma and brain disposition and pharmacodynamics of methadone in abstinent rats. AB - The pharmacokinetics and pharmacodynamics of methadone were investigated in control and abstinent rats. Minipumps filled with saline (control group) or saline-morphine (abstinent group) solutions were used to induce physical dependence. Solutions were delivered continuously by minipumps for 6 days. The physical dependence was evaluated 12 h after minipump removal by measuring specific withdrawal signs. Animals from the abstinent group showed clear withdrawal signs such as hostility on handling and weight loss. Plasma and brain disposition and pharmacodynamics of methadone were evaluated after a 0.35 mg/kg i.v. bolus dose administered 12 h after minipump removal. Plasma clearance, distribution clearance, and volume of distribution at steady-state were significantly decreased (P < 0.05) in the abstinent group. Plasma levels of alpha1-acid glycoprotein and plasma protein binding were significantly increased (P < 0.05) in the abstinent group. The estimates of pharmacokinetic parameters based on unbound plasma concentrations did not differ between groups, with the sole exception of the unbound apparent volume of distribution. The access of methadone to the brain was significantly faster (P < 0.05) in the abstinent group, although the extent of distribution in the brain was diminished in comparison with the control group. Analgesia recorded with tail-flick was used as the pharmacodynamic endpoint. Analgesic response and effect compartment concentrations of methadone were related by the sigmoidal Emax model. Estimates of C50 [steady-state plasma concentrations eliciting half of maximum effect (Emax)]] based on unbound concentrations did not differ between groups. On the other hand, the estimate of Emax had decreased by 65% in the abstinent group. PMID- 9862770 TI - Subjective and physiological effects of intravenous nicotine and cocaine in cigarette smoking cocaine abusers. AB - The subjective and physiological effects of intravenously administered cocaine and nicotine were compared in 10 cigarette-smoking cocaine abusers. Subjects abstained from smoking at least 8 h before each session. Under double blind conditions, subjects received placebo, cocaine (10, 20, and 40 mg/70 mg), or nicotine (0.75, 1.5, 3.0 mg/70 kg) in mixed order. Physiological and subjective data were collected before and repeatedly after each intravenous drug administration. Subjects also completed a drug versus money multiple-choice procedure in which they chose between that day's drug and 44 monetary values. Both drugs increased blood pressure and heart rate and decreased skin temperature. Nicotine showed a more rapid onset of subjective effects than cocaine. Overall, although both cocaine and nicotine increased subjective ratings of "drug effect", "rush", "good effects", "liking", "high", and "stimulated", only nicotine increased ratings of "bad effects" and "jittery". Although the highest nicotine dose produced greater effects than the highest cocaine dose on most subjective measures, the highest cocaine dose produced somewhat greater ratings of drug liking. At doses that produced comparable ratings of drug effect (40 mg/70 kg cocaine versus 1.5 mg/70 kg nicotine), cocaine produced significantly greater good effects, whereas nicotine produced greater bad effects. All three cocaine doses and the intermediate and high nicotine doses were frequently categorized as producing effects similar to those of cocaine or amphetamine. The drug versus money measure showed that the highest cocaine dose was worth twice as much as the highest nicotine dose. Thus, intravenous cocaine and nicotine can be differentiated by their subjective and reinforcing effects. PMID- 9862771 TI - High-affinity efflux transport system for glutathione conjugates on the luminal membrane of a mouse brain capillary endothelial cell line (MBEC4). AB - Cumulative evidence suggests that several organic anions are excreted from the brain to the blood across the blood-brain barrier. In the present study, we carried out a kinetic investigation of the transport activity in MBEC4, an immortalized cell line established from BALB/c mouse cerebral microvessel endothelial cells. The presence of an efflux system in intact cells was examined by using monochlorobimane (MCB), which is conjugated with glutathione intracellularly to produce glutathione bimane (GS-B). The efflux of GS-B was inhibited by ATP depletion and also by 1-chloro-2, 4-dinitrobenzne, a precursor of 2,4-dinitrophenyl-S-glutathione, in a concentration-dependent manner. Using this MBEC4 monolayer, we investigated the direction of this transport activity. Although the efflux of GS-B was observed on both luminal and abluminal sides of MBEC4 monolayer, the profile differed for the two sides with respect to the concentration dependence of MCB; the analysis suggested the presence of high affinity transport system on the luminal side. To investigate the mechanism for the transport, we examined the ATP-dependent uptake of GS-B into the membrane vesicles prepared from MBEC4. ATP-dependent uptake systems with high (Km = 35 nM) and low (Km = 14 microM) affinities were identified. These results suggested that this high-affinity transport system of glutathione conjugates is expressed on the luminal side of the blood-brain barrier and is involved in the detoxification of xenobiotics. PMID- 9862772 TI - Differences in degree of trapping of low-affinity uncompetitive N-methyl-D aspartic acid receptor antagonists with similar kinetics of block. AB - This study characterizes the trapping of block of N-methyl-D-aspartic acid (NMDA) induced currents by three structurally distinct, use-dependent NMDA receptor antagonists with similar rapid on-off rates. The antagonism of whole-cell currents in cultured rat cortical neurons by AR-R15896AR, ketamine, and memantine was examined. All three compounds produced a steady-state block after a 30-s coapplication, which was fully relieved after 50 s of NMDA exposure. The amplitudes of block caused by 50 microM AR-R15896AR, 10 microM ketamine, or 10 microM memantine were not significantly different, being 82 +/- 1%, 80 +/- 2%, and 81 +/- 2%, respectively. All three NMDA receptor antagonists exhibited trapping of block that was not significantly increased by extending the agonist/antagonist coapplication beyond 30 s. Although the initial blocks were similar, after 120 s of washout without agonist present, there were significant differences in trapping of block between antagonists, as only 54 +/- 3% of the AR R15896AR block, 86 +/- 1% of the ketamine block, and 71 +/- 4% of the memantine block remained trapped. The lack of complete trapping is consistent with closed channel egress by these compounds. Higher antagonist concentrations produced larger initial blocks, but the degree of trapping block was not significantly different from that at lower antagonist concentrations. The results demonstrate that differences in the degree of trapping exist among use-dependent NMDA receptor antagonists even when on and off rates are similar. These differences are correlated with measures of therapeutic index. PMID- 9862773 TI - Potentiation of ethanol effects in cerebellum by activation of endogenous noradrenergic inputs. AB - We previously found that beta adrenergic agonists such as norepinephrine and isoproterenol potentiate the depressant actions of ethanol (EtOH) on cerebellar Purkinje neurons. Furthermore, antagonism of the beta adrenergic effects of endogenously released catecholamines with timolol reduced EtOH-induced depressions of neuronal activity in that brain area. In the present study, we further investigated the hypothesis that activity of the endogenous noradrenergic innervation to the cerebellar cortex can potentiate this EtOH action. We investigated the interaction of synaptically released catecholamines on EtOH induced depressions of cerebellar Purkinje neurons in three different experiments: (1) endogenous catecholamine release was facilitated by applying the catecholamine uptake inhibitor desmethylimipramine, (2) activity of the noradrenergic innervation of the cerebellar cortex from locus ceruleus was increased by causing acute withdrawal from 7 days of chronic morphine treatment with the opiate antagonist naloxone, and (3) the noradrenergic innervation of the cerebellum was activated directly by electrical stimulation of the locus ceruleus. We found that all three conditions potentiated EtOH-induced depressions in the cerebellum and that this potentiation of ethanol effects could be antagonized by the systemic administration of the beta adrenergic antagonist propranolol. Furthermore, morphine withdrawal also caused potentiation of the depressant effects of phencyclidine, which are known to be regulated by the endogenous catecholamine innervation in this brain area. Taken together with our previous data demonstrating a beta adrenergic facilitation of EtOH actions in this brain area, the present results suggest that the activity of endogenous noradrenergic synapses can regulate the depressant effects of EtOH on cerebellar Purkinje neurons. PMID- 9862774 TI - Characterization of (S)-des-4-amino-3-[125I]iodozacopride ([125I]DAIZAC), a selective high-affinity radioligand for 5-hydroxytryptamine3 receptors. AB - The 5-hydroxytryptamine(HT)3 receptor subtype is present in the central nervous system (CNS) in low abundance, and few selective radiolabeled antagonists with high specific activity are available to study these sites. DAIZAC [desamino-3 iodo-(S)-zacopride; (S)-5-chloro-3-iodo-2-methoxy-N-(1-azobicyclo-[2.2. 2]oct-3 yl)benzamide] is a compound with high affinity and selectivity for the 5-HT3 receptor. Scatchard analysis of specific binding to NCB-20 cell membranes gave a Bmax of 340 +/- 58 fmol/mg protein and a KD of 0.14 +/- 0.03 nM, which is in agreement with the value previously reported in rat brain (KD = 0.15 nM). Nonspecific binding of [125I]DAIZAC in NCB-20 cells was <1% of total binding at the KD for DAIZAC compared with 17% in the rat brain preparation. Unlabeled DAIZAC (10 microM) showed minimal ability to displace binding of radiolabeled ligands selected for their affinities for other CNS receptor and uptake carrier binding sites. The discrimination ratio of DAIZAC for the 5-HT3 receptor over the M1 muscarinic binding site, the non-5-HT3 site at which it was most potent, was >2800. Serotonergic antagonists at every other known CNS serotonergic binding sites (3-30 microM) were ineffective in displacing [125I]DAIZAC binding in rat brain membranes. Similarly, antagonists (3-30 microM) for other nonserotonergic receptors and uptake sites were ineffective in displacing [125I]DAIZAC binding. Autoradiographic studies showed highest specific binding in area postrema and nucleus solitarius, with intermediate levels of binding in entorhinal cortex and hippocampus. DAIZAC inhibited 5-HT3 receptor-mediated inward cation current in NCB-20 cells with an IC50 of 0.24 nM. [125I]DAIZAC is a potent and highly selective ligand for in vitro studies of the 5-HT3 receptor. PMID- 9862775 TI - GalphaL1 (Galpha14) couples the opioid receptor-like1 receptor to stimulation of phospholipase C. AB - In most tissues and cells the opioid receptor-like (ORL1) receptor regulates effectors primarily through the pertussis toxin (PTX)-sensitive guanine nucleotide-binding regulatory proteins (G proteins) Gi/Go. Many Gi-coupled receptors possess additional capability to interact with one or more PTX insensitive G proteins. Using the betagamma-induced stimulation of type 2 adenylyl cyclase as a readout, we screened the ability of ORL1 receptor to interact with a panel of PTX-insensitive G proteins. In the presence of PTX, activation of the ORL1 receptor resulted in the stimulation of type 2 adenylyl cyclase only in HEK 293 cells coexpressing the alpha subunit of Gz, G12, G14, or G16, but not in cells coexpressing G11, G13, or Gq. Coupling to both Gz and G16 was expected because close relatives of the ORL1 receptor, the opioid receptors, are known to couple productively to these G proteins. ORL1 receptor coupling to either G12 or G14 has not been demonstrated. As predicted by the type 2 adenylyl cyclase assays, activation of the ORL1 receptor resulted in the formation of inositol phosphates in COS-7 cells transiently cotransfected with Galpha14. The ORL1 receptor-mediated stimulation of phospholipase C was found to be Galpha14 dependent, agonist dose dependent, ligand selective, and PTX insensitive. We conclude that G14 can link the ORL1 receptor to regulation of phopholipase C. PMID- 9862776 TI - Mechanisms and sites of action of endothelins 1 and 2 on the opossum internal anal sphincter smooth muscle tone in vitro. AB - Endothelins, localized in the enteric nervous system, may play important roles in the morphogenesis of the gastrointestinal (GI) tract and in the regulation of GI motility. However, the role of endothelins in the GI sphincters, including the internal anal sphincter (IAS) have not been examined. We examined the actions of endothelins on the basal tone of the opossum IAS circular smooth muscle strips before and after different neurohumoral antagonists or inhibitors. Endothelins 1 and 2 produced a concentration-dependent biphasic effect on the basal tone of the IAS, an initial brief fall followed by a sustained rise. The fall in the IAS smooth muscle tone was not modified by atropine, guanethidine, or tetrodotoxin but was significantly attenuated by the nitric oxide synthase inhibitor L-NNA, the specific neuronal nitric oxide synthase inhibitor, 1-(2 trifluoromethylphenyl)imidazole, the N-type neuronal Ca++-channel blocker omega conotoxin GVIA, and by the calmodulin antagonist W-13. Endothelin-induced contraction of the IAS, on the other hand, was not affected by any of the neurohumoral antagonists but was significantly inhibited by the selective protein kinase C inhibitor H-7 or the calmodulin inhibitor W-13. The combination of H-7 and W-13 had no additive effect in attenuating the contractile action of endothelin 1. There was clear evidence of a cross-tachyphylaxis to the actions of endothelin 1 and endothelin 2. We conclude that the endothelins exert important neuromodulatory effects on the basal tone of the IAS. The contractile action occurs directly at the smooth muscle and the relaxant action by the activation of neuronal nitric oxide synthase at the nerve terminals. The contraction and relaxation of the smooth muscle caused by endothelins 1 and 2 may involve distinct receptors that are similar for both endothelins. The excitatory actions of endothelin 1 involve both the protein kinase C and the Ca++-calmodulin pathways that may lie in series. PMID- 9862777 TI - Long-term sequential determination of behavioral ontogeny of 5-HT1A and 5-HT2 receptor functions in the rat. AB - Activation of 5-hydroxytryptamine1A (5-HT1A) receptors in rats produces hypothermia and a number of behaviors [hindleg abduction (HLA), lateral head weaving (LHW), forepaw treading (FPT), flat body posture (FBP), rollover (RO), tremor (T), and straub tail (ST)] known collectively as the serotonin syndrome (SS). Stimulation of 5-HT2A receptors produces wet-dog shakes (WDS), whereas 5 HT2C sites induce back muscle contraction (BMC). We investigated the functional ontogeny of the cited receptors in rat pups on postnatal days (PD) 7, 14, 18, 22, 28, 35, 60, and 120 by using (1) the 5-HT1A agonist 8-hydroxy-2 dipropylaminotetralin (0, 1.25, and 5 mg/kg) to induce the SS and hypothermia and (2) the 5-HT2A/C agonist (+/-)-1-(2, 5-dimethoxy-4-iodophenyl)-2-aminopropane (0, 0.5, and 4 mg/kg) to produce both WDS and BMC. The age of onset for most symptoms of SS [FBP, HLA, RO, and T] was the first week of life. They attained maximal intensities at ages 7 to 14 days, after which their maxima either reduced or dissipated to zero. Per contra, the onset of LHW and FPT required 14 to 18 days, and their maxima developed later. The onset of (+/-)-1-(2, 5-dimethoxy-4 iodophenyl)-2-aminopropane-induced WDS occurred after PD 14, and by PD 18, it reached its maximal intensity, which persisted up to PD 60, after which it declined. The onset of BMC was evident on PD 28 and attained its maximal frequency at ages 90 to 120 days. The results show that different components of SS appear within 14 days of birth, but they mature differentially, whereas the hypothermic effect of 5-HT1A receptors remains relatively constant during aging. The times of onset and maturation of WDS were intermediate (between the second and third weeks of life), whereas BMC required 1 to 2 months for its appearance and maturation. PMID- 9862778 TI - Peroxisomes are involved in the swift increase in alcohol metabolism. AB - The purpose of this study was to determine whether catalase-dependent alcohol metabolism is activated by alcohol (i.e., swift increase in alcohol metabolism). When ethanol or the selective substrate for catalase, methanol, was given (5.0 g/kg) in vivo 2 to 3 h before liver perfusion, methanol and oxygen metabolism were increased significantly. This increase was blocked when the specific Kupffer cell toxicant GdCl3 was administered 24 h before perfusion. These data support the hypothesis that catalase-dependent alcohol metabolism is activated by acute alcohol and that Kupffer cells are involved. Ethanol treatment in vivo increased ketogenesis from endogenous fatty acids nearly 3-fold and increased plasma triglycerides and hepatic acyl CoA synthetase activity; all increases were blocked by GdCl3. These findings support the hypothesis that ethanol increases H2O2 supply for catalase-dependent alcohol metabolism by increasing fatty acid supply. Infusion of oleate stimulated oxygen uptake 1.5-fold and methanol metabolism 4-fold, but these parameters were not altered by GdCl3. Moreover, the effects of ethanol treatment were blocked by the cyclooxygenase inhibitor indomethacin, and prostaglandin E2 (PGE2) was increased more than 200% in media from cultured Kupffer cells from rats treated with ethanol in vivo. Furthermore, lipoprotein lipase activity in retroperitoneal fat pads, which is known to be inhibited by PGE2, was reduced 70% by ethanol. These data are consistent with the hypothesis that Kupffer cells play a key role in activation of catalase-dependent alcohol metabolism, most likely by producing mediators (e.g., PGE2) that inhibit lipoprotein lipase, increase the supply of fatty acids to the liver, and increase generation of H2O2 via peroxisomal beta-oxidation. PMID- 9862779 TI - Dynorphin A1-13 causes elevation of serum levels of prolactin through an opioid receptor mechanism in humans: gender differences and implications for modulation of dopaminergic tone in the treatment of addictions. AB - Dynorphin peptides act preferentially at kappa- as well as mu- and delta-opioid receptors. This study was conducted to determine whether dynorphin peptides act to lower dopaminergic tone in the tuberoinfundibular system, resulting in elevated serum prolactin levels and, if so, whether such an effect is mediated by the opioid receptors. Dose-related increases in serum prolactin levels were observed after dynorphin A1-13 was administered i.v. in doses of 120 and 500 micrograms/kg to healthy human volunteers with no history of drug or alcohol abuse. Studies were then conducted to determine whether this effect is opioid receptor mediated and, if so, whether at kappa- or mu types. Pretreatment with the opioid antagonist nalmefene (30 mg i.v.), which has high affinity at both mu- and kappa-opioid receptors, caused a greater attenuation in dynorphin A1-13 stimulated increases in serum prolactin levels than pretreatment with similarly high doses of naloxone, an antagonist with lower affinity for both mu- and kappa opioid receptors. These results suggest dynorphin A1-13 lowers tuberoinfundibular dopaminergic tone through action at kappa- and possibly mu-opioid receptors. Female subjects were significantly more responsive to the prolactin effects of dynorphin than were male subjects. Dynorphin gene expression, dynorphin peptides, and kappa-opioid receptor gene expression and binding have been shown to be altered in response to cocaine administration. Also, both dynorphin peptides and synthetic kappa-opioid agonists have been shown to lower dopamine levels in the nucleus accumbens and to attenuate cocaine-induced surges in dopamine levels. Thus, a dynorphin-like compound capable of reaching critical mesolimbic mesocortical and nigrostriatal dopaminergic systems may be effective in the management of cocaine addiction. PMID- 9862780 TI - The in vivo unidirectional conversion of nitro-D-arginine to nitro-L-arginine. AB - We recently reported that nitric oxide synthase in the brain can be inhibited not only by nitro-L-arginine (L-NA) but also by its D-enantiomer nitro-D-arginine (D NA). In the present study, we found that D-NA, when tested in vitro, was 400 times less potent than L-NA. However, when D-NA was injected in vivo, its L enantiomer, L-NA, was found to rapidly appear in plasma samples (approximately 1 min), rose to a maximum concentration at 30 min (approximately 40% conversion), and remained at this plateau for about 5 h. This was consistent with the changes in blood pressure. There was no conversion of L- to D-NA. The results suggested that D-NA has very weak biological actions by itself, but when administered in vivo, D-NA can be converted to L-NA. PMID- 9862781 TI - Synergistic elevations in nucleus accumbens extracellular dopamine concentrations during self-administration of cocaine/heroin combinations (Speedball) in rats. AB - The abuse of cocaine/opiate combinations (speedball) represents a growing trend in illicit drug use. Delineation of neurobiological substrates mediating the reinforcing effects of the combination may increase our knowledge of reinforcement mechanisms and provide useful new information for the development of pharmacotherapies. Several studies suggest dopaminergic innervations of the nucleus accumbens (NAc) have a central role in the brain processes underlying drug reinforcement. The present study was undertaken to determine the relationship between the self-administration of cocaine/heroin combinations and NAc extracellular dopamine concentrations ([DA]e) using in vivo microdialysis and microbore high-pressure liquid chromatography. Rats were assigned randomly to one of three groups to self-administer i.v. cocaine (125, 250, and 500 micrograms/infusion; n = 5), heroin (4.5, 9, and 18 micrograms/infusion; n = 5), or cocaine/heroin combinations (125/4.5; 250/9, and 500/18 micrograms/infusion; n = 4) under a fixed ratio (FR) 10: 20-s time-out schedule of reinforcement/multicomponent dosing session. After stable rates of responding were engendered and maintained, microdialysis samples were collected in 10-min intervals during the self-administration session. Self-administration of cocaine/heroin combinations produced synergisitic elevations in NAc [DA]e (1000% baseline) compared with cocaine (400% baseline) and heroin (not significantly different from baseline levels). Neither the number of infusions nor the interinfusion intervals was significantly different between the groups across the self-administration session. Moreover, cocaine concentrations were not significantly different between the cocaine and cocaine/heroin groups. These results demonstrate that heroin interacts with cocaine to produce synergistic elevations in [DA]e, providing a neurochemical basis for understanding the abuse liability of cocaine/opiate combinations. PMID- 9862782 TI - Specific delivery of captopril to the kidney with the prodrug captopril-lysozyme. AB - Low-molecular-weight proteins (LMWPs) accumulate in the proximal tubular cells of the kidney, which makes these proteins interesting tools for renal drug targeting. We studied this approach using the LMWP lysozyme as a carrier for the angiotensin-converting enzyme inhibitor captopril. Captopril was conjugated to lysozyme via a disulfide bond. The pharmacokinetics of the captopril-lysozyme conjugate was studied in the rat. Only intact conjugate could be detected in the circulation. The total amount of captopril disulfides in the kidney was six times higher after administration of the conjugate than after the administration of an equivalent amount of free captopril. The conjugate was recovered in the urine partially as intact conjugate and partially as low-molecular-weight disulfides. The excretion of conjugate in the urine was not a consequence of the coupling of captopril to lysozyme because an equivalent bolus dose of native lysozyme was similarly excreted into the urine. By determination of the renal angiotensin converting enzyme activity, we showed that the conjugate was degraded to the pharmacologically active captopril in vivo. We conclude that the coupling of captopril to the LMWP lysozyme results in increased captopril concentrations in the kidney and reduced captopril concentrations in the circulation. PMID- 9862783 TI - Pharmacologic actions of the second-generation leukotriene B4 receptor antagonist LY293111: in vitro studies. AB - The in vitro actions were investigated of LY293111, a potent and selective leukotriene B4 (LTB4) receptor antagonist, on human neutrophils, human blood fractions, guinea pig lung membranes, and guinea pig parenchymal and tracheal strips. The IC50 for inhibiting [3H]LTB4 binding to human neutrophils was 17.6 +/ 4.8 nM. LY293111 inhibited LTB4-induced human neutrophil aggregation (IC50 = 32 +/- 5 nM), luminol-dependent chemiluminescence (IC50 = 20 +/- 2 nM), chemotaxis (IC50 = 6.3 +/- 1.7 nM), and superoxide production by adherent cells (IC50 = 0.5 nM). Corresponding responses induced by N-formyl-L-methionyl-L-leucyl-L phenylalanine were inhibited by 100-fold higher concentrations of LY293111. LTB4 binding to guinea pig tissues and subsequent activation were also inhibited. The Ki for inhibition of [3H]LTB4 binding to lung membranes was 7.1 +/- 0.8 nM; IC50 for preventing binding of [3H]LTB4 to spleen membranes was 65 nM. The compound inhibited LTB4-induced contraction of guinea pig lung parenchyma. At 10 nM, LY293111 caused a parallel rightward shift of the LTB4 concentration-response curve. At higher concentrations, plots were shifted in a nonparallel manner, and maximum responses were depressed. LY293111 did not prevent antigen-stimulated contraction of sensitized trachea strips. At micromolar concentrations, LY293111 inhibited production of LTB4 and thromboxane B2 by plasma-depleted human blood stimulated with N-formyl-L-methionyl-L-leucyl-L-phenylalanine and thrombin. In addition, at these higher concentrations, formation of LTB4 by A23187-activated whole blood and conversion of arachidonic acid to LTB4 by a human neutrophil cytosolic fraction were inhibited. In summary, LY293111 is a second-generation LTB4 receptor antagonist with much improved potency in a variety of functional assay systems. PMID- 9862784 TI - Effects of alpha- and beta-adrenoceptor blockade on purine secretion induced by sympathetic nerve stimulation in the rat kidney. AB - To characterize the effects of renal sympathetic nerve activation (RSNA) on renal purine secretion, 13 perfused rat kidneys were stimulated with periarterial electrodes at 7 Hz for 3 min, and purine secretion was determined by measuring with high-performance liquid chromatography purines in the renal venous perfusate 1 min before and during the last minute of RSNA. RSNA significantly increased renal perfusion pressure and significantly increased the secretion of adenosine and adenosine metabolites (inosine, hypoxanthine, and xanthine) by 2- to 5-fold. To investigate the participation of alpha- and beta-adrenoceptors in this response, four groups of perfused kidneys (n = 5/group) were pretreated with either vehicle, prazosin (alpha1-adrenoceptor antagonist; 0.03 microM), phentolamine (alpha1/2-adrenoceptor antagonist; 3 microM), or propranolol (beta1/2-adrenoceptor antagonist; 0.1 microM), and purine secretion was measured before and during RSNA at 1, 3, 5, 7, and 9 Hz. Prazosin, phentolamine, and propranolol abolished the RSNA-induced increase in the secretion of adenosine, inosine, hypoxanthine, and xanthine. In contrast, prazosin and phentolamine nearly abolished, whereas propranolol only slightly reduced, renal vascular responses to RSNA. Our results indicate that RSNA increases renal purine secretion via a mechanism that requires both alpha- and beta-adrenoceptors. It is well known that in the kidney adenosine activates renal afferent nerves, enhances renovascular responses to norepinephrine and angiotensin II, and increases sodium reabsorption; therefore, RSNA-induced adenosine production may contribute to the hypertensive effects of RSNA. Moreover, the antihypertensive effects of beta adrenoceptor antagonists may in part be due to inhibition of RSNA-induced renal adenosine production. PMID- 9862785 TI - Novel 3alpha-diphenylmethoxytropane analogs: selective dopamine uptake inhibitors with behavioral effects distinct from those of cocaine. AB - The pharmacological effects were assessed for a series of 3alpha-diphenylmethoxy 1alphaH,5alphaH-tropane analogs which have structural similarities to cocaine. Like cocaine, these compounds displaced [3H]WIN 35,428 binding from rat caudate and had affinities ranging from approximately 10-fold greater than cocaine (Ki=11.8 nM) to relatively low affinity (Ki=2000 nM). The compounds also inhibited dopamine uptake with potencies corresponding to their affinities for WIN 35,428 binding sites. Like the parent compound, benztropine, the 3alpha (diphenylmethoxy)tropane analogs displaced [3H]pirenzepine from muscarinic M1 receptors with affinities ranging from 2 to 120 nM. Cocaine produced dose-related increases in locomotor activity (horizontal ambulation) in Swiss Webster mice, whereas the 3alpha-(diphenylmethoxy)tropane analogs generally had lower efficacy than cocaine. Compounds with fluoro-substituents in the phenyl rings generally were among those with efficacy approaching that of cocaine; those with chloro- and bromo-substituents were markedly less efficacious, despite having binding affinities comparable to those of the corresponding fluoro-substituted compounds. The 3alpha-(diphenylmethoxy)tropane analogs were also examined in rats trained to discriminate saline from cocaine (10 mg/kg, i.p.). Cocaine produced a dose related increase in responding on the cocaine-appropriate lever, reaching 100% at 10 mg/kg. Only the 4',4"-difluoro-substituted analog produced effects similar to those of cocaine; the other compounds showed markedly reduced efficacy compared to cocaine. Drug interaction studies showed that the antimuscarinics, atropine and scopolamine, potentiated rather than attenuated the locomotor stimulant and cocaine-like discriminative-stimulus effects of cocaine, indicating that the antimuscarinic effects of the 3alpha-diphenylmethoxytropane analogs did not contribute to their diminished cocaine-like activity. Studies of the time course of selected compounds indicated that their reduced cocaine-like efficacy was likely not due to behavioral observations being conducted at an inopportune time period. Because none of the 3alpha-diphenylmethoxytropane analogs studied showed evidence that they were binding to more than one site, and because the structure activity relationships among these drugs are distinctly different from those obtained with cocaine, these data suggest that the 3alpha-diphenylmethoxytropane analogs are accessing a different binding domain than that accessed by cocaine. Binding to this domain may produce a behavioral profile that is distinct from that of the cocaine-like dopamine uptake inhibitors. PMID- 9862786 TI - Levosimendan, a calcium sensitizer in cardiac muscle, induces relaxation in coronary smooth muscle through calcium desensitization. AB - Levosimendan is a pyridazinone-dinitrile derivative belonging to a new class of cardiac inotropic drugs, Ca++ sensitizers. Levosimendan is also a vasodilator both in vitro and in vivo, but its mechanism is not well understood. The cardiac target protein of levosimendan, troponin C, is a Ca++-binding EF-hand protein. This raises the possibility that levosimendan may also interact with smooth muscle EF-hand proteins, such as, calmodulin, the regulatory myosin light chains, or S100 proteins. We investigated the effects of levosimendan on [Ca++]i, and force in porcine coronary arteries, with receptor-mediated (U46619) or KCl stimulation. At high levels of stimulation, levosimendan decreased force without changing or increasing [Ca++]i, measured with the Ca++-sensitive fluorescent probe fura-2 in the intact artery. With lower levels of U46619, levosimendan (1 microM) lowered force by 70% and reduced [Ca++]i by 38%. The relationship between force and [Ca++]i for KCl stimulation are significantly rightward shifted, indicating Ca++ desensitization by levosimendan. In contrast, the phosphodiesterase III inhibitor, milrinone, does not shift the force-Ca++ relations but elicits relaxation via lowering [Ca++]i. There was little change in pHi, indicating that the Ca++ desensitization by levosimendan was not attributable to decreasing pHi. Levosimendan relaxes coronary arteries and lowers [Ca++]i by mechanisms different than milrinone. Our results indicate a lowering of [Ca++]i by levosimendan consistent with opening of potassium channels and a relaxation that is independent of [Ca++]i. Our evidence points to a novel mechanism that might involve the direct effect of levosimendan on the smooth muscle contractile or regulatory proteins themselves. PMID- 9862787 TI - Metabolic transformations of leukotriene B4 in primary cultures of human hepatocytes. AB - Leukotriene B4 (LTB4) is a potent lipid mediator of the inflammatory response whose biological half-life is believed to be mediated principally by metabolism to inactive forms either in the tissue of origin or in the liver. Pathways of metabolic degradation of LTB4 along with structural identification of metabolites have been elucidated previously in isolated rat liver cells, human keratinocytes, human polymorphonuclear leukocytes, and cultured HepG2 cells. Research advances in human liver transplantation and preservation have made isolated human hepatocytes available for studying the metabolism of LTB4 in vitro. LTB4 was added to plated human hepatocytes from three different subjects for 24-h periods whereupon the substrate was analyzed by high-performance liquid chromatography coupled with scintillation counting, UV spectroscopy, and negative ion electrospray ionization tandem mass spectrometry. Each set of hepatocytes yielded a different distribution of metabolites, but several metabolites appeared in all three sets of cells. These central metabolites included the previously identified 20-carboxy-LTB4 and 18-carboxy-LTB4, implicating the presence in the liver of specific P-450-mediated omega-oxidation as well as the enzymes involved in beta oxidation from the omega-terminus. Each set of hepatocytes produced the metabolite 10,11-dihydro-20-COOH-LTB4, a product of the 12-hydroxyeicosanoid dehydrogenase/Delta10 reductase pathway. Glucuronides of LTB4 and several metabolites were found, which represents the first description of glucuronidation as a pathway of LTB4 metabolism. Finally, a series of novel metabolites were observed corresponding to beta-oxidation from the carboxyl terminus of LTB4. PMID- 9862788 TI - Full and partial 5-HT1A receptor agonists disrupt learning and performance in rats. AB - As a means of characterizing the role of 5-hydroxytryptamine (5-HT1A) receptors in learning, a full 5-HT1A receptor agonist, 8-hydroxy-dipropylaminotetralin (8 OH-DPAT), was administered both alone and in combination with two partial agonists (buspirone and 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine hydrobromide (NAN-190)) and a 5-HT1A receptor antagonist (p-MPPI) to rats responding under a multiple schedule of repeated acquisition and performance of response sequences. In addition, the effects of another 5-HT1A receptor agonist, (LY228729), were also studied under this same procedure. When administered alone, both 8-OH-DPAT (0.1-3. 2 mg/kg) and LY228729 (0.32-3.2 mg/kg) dose dependently decreased overall response rate and increased the percentage of errors in the acquisition and performance components. At the doses of each drug tested, both buspirone (0.32 or 1 mg/kg) and NAN-190 (1 or 3.2 mg/kg) also decreased overall response rate and increased the percentage of errors. However, the effects of these drugs differed across behavioral components and dependent measures. The effects of buspirone and NAN-190 on rate and accuracy were also different when they were administered in combination with 8-OH-DPAT. In contrast, p-MPPI (3.2 or 10 mg/kg) had little or no effect when administered alone and antagonized the effects of 8-OH-DPAT; shifting the dose-effect curves for both response rate and the percentage of errors in both components to the right. Taken together, these results indicate that complex behaviors in rats are sensitive to disruption by drugs with both full and partial 5-HT1A receptor agonist properties, and that the effects of partial 5-HT1A receptor agonists on learning may be different depending on their efficacy at pre- and postsynaptic 5-HT1A receptors. PMID- 9862789 TI - Characterization of binding properties to human P-glycoprotein: development of a [3H]verapamil radioligand-binding assay. AB - Interaction with the exsorptive transporter P-glycoprotein (P-gp) is a possible source of peculiarities in drug pharmacokinetics, including dose-dependent absorption, drug-drug interactions, intestinal secretion, and limited permeability of the blood-brain barrier. Among the established in vitro methods of the analysis of drug interactions with P-gp, none directly quantifies the affinity of ligands with P-gp. Instead, they measure the result of a membrane permeation and a receptor-binding process; this may lead to difficulties in the interpretation of results. An assay for quantification of drug affinity to the transporter is presented on the basis of the radioligand-binding assay principle. This has the advantage of directly quantifying the interaction between drugs and P-gp. Because of the reversible and competitive interaction of numerous substrates with P-gp, a radioligand-binding assay was developed by taking [3H]verapamil and [3H]vinblastine as radioligands and the human intestinal Caco-2 cells, overexpressed with P-gp by culturing in the presence of vinblastine or transfecting with multidrug resistance gene MDR-1 as receptor preparation. The assay was performed in 96-well plates and has the potential to be used as a high throughput method. A clear induction of the expression of P-gp was demonstrated in the Caco-2 cells grown in the presence of vinblastine, as well as in the transfected cells, although to a lesser extent. Both radioligands were shown to bind to P-gp. Verapamil was the radioligand of choice for further investigations due to its lower nonspecific binding to the transporter preparation. Kinetics as well as specificity of the binding of verapamil to the P-gp preparation were demonstrated. A two-affinity model was found to adequately describe the data derived from saturation as well as from competition experiments, in accordance with previous findings on two exsorption sites for P-gp. The binding properties of [3H]verapamil and [3H]vinblastine to a P-gp preparation derived from induced Caco-2 cells are described. The concentration-dependent displacement of the radioligand by nonlabeled substrates for P-gp should be a suitable principle for the determination of drug affinity to the respective binding sites at the human intestinal multidrug transporter P-gp. PMID- 9862790 TI - Evaluation of proteinase-activated receptor-1 (PAR1) agonists and antagonists using a cultured cell receptor desensitization assay: activation of PAR2 by PAR1 targeted ligands. AB - We developed a calcium signaling-based assay, using cultured human embryonic kidney cells (HEK), that evaluates simultaneously, the activation/desensitization or blockade of the proteinase-activated receptors, PAR1 and PAR2. Using this assay, we analyzed the actions of a number of previously described putative PAR1 targeted peptide agonists and antagonists. We found that most of the previously described PAR1-targeted agents can also activate/desensitize PAR2, and most of these peptides can also activate a calcium signaling pathway in a target cell that possesses PAR2 along with PAR1. Furthermore, we used this assay to develop a PAR1 receptor-activating probe [Ala-parafluoroPhe-Arg-Cha-Cit-Tyr-NH2 (Cit-NH2)], which displays a high degree of specificity for PAR1 over PAR2, and we used the assay to quantitate the ability of trypsin to disarm the activation of PAR1 by thrombin. The abilities of the PAR1-targeted agents to desensitize or block PAR1 in the HEK cell assay were compared with their activities in a human platelet aggregation assay. Our data illustrate the usefulness of the HEK cell assay for evaluating the PAR1/PAR2 selectivity of PAR-activating agonists. The PAR1 selective agonist that we developed using the assay should prove useful for studying the effects of selectively activating PAR1 in vivo. PMID- 9862792 TI - Differential actions of pacific ciguatoxin-1 on sodium channel subtypes in mammalian sensory neurons. AB - Pacific ciguatoxin-1 (P-CTX-1), is a highly lipophilic cyclic polyether molecule originating from the marine dinoflagellate Gambierdiscus toxicus. Its effects were investigated on sodium channel subtypes present in acutely dissociated rat dorsal root ganglion neurons, using whole-cell patch clamp techniques. Concentrations of P-CTX-1 ranging from 0.2 to 20 nM had no effect on the kinetics of tetrodotoxin-sensitive (TTX-S) or tetrodotoxin-resistant (TTX-R) sodium channel activation and inactivation, however, a concentration-dependent reduction in peak current amplitude occurred in both channel types. The main actions of 5 nM P-CTX-1 on TTX-S sodium channels were a 13-mV hyperpolarizing shift in the voltage dependence of sodium channel activation and a 22-mV hyperpolarizing shift in steady-state inactivation (hinfinity). In addition, P-CTX-1 caused a rapid rise in the membrane leakage current in cells expressing TTX-S sodium channels. This effect was blocked by 200 nM TTX, indicating an action mediated through TTX S sodium channels. In contrast, the main action of P-CTX-1 (5 nM) on TTX-R sodium channels was a significant increase in the rate of recovery from sodium channel inactivation. These results indicate that P-CTX-1 acts to modify voltage-gated sodium channels present in peripheral sensory neurons consistent with its action to increase nerve excitability. This provides an explanation for the sensory neurological disturbances associated with ciguatera fish poisoning. PMID- 9862791 TI - Preclinical pharmacokinetics, interspecies scaling, and tissue distribution of a humanized monoclonal antibody against vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) plays a crucial role in angiogenesis and in pathological processes such as tumor growth, rheumatoid arthritis, and ocular neovascularization. A recombinant humanized monoclonal antibody (rhuMAb), rhuMAb VEGF, has been developed to inhibit the effects of VEGF in the treatment of solid tumors. Intravenous and s.c. pharmacokinetic studies were conducted in mice, rats, and cynomolgus monkeys. In addition, the tissue distribution of i.v. 125I-rhuMAb VEGF was investigated in rabbits. At a dose of approximately 10 mg/kg, the clearance of rhuMAb VEGF from the serum was 15.7 ml/day/kg in mice, 4.83 ml/day/kg in rats, and 5.59 ml/day/kg in cynomolgus monkeys, and the terminal half-life ranged from 6 to 12 days in all species. After s.c. administration, rhuMAb VEGF had a bioavailability of 69% in rats and 100% in mice and cynomolgus monkeys. Pharmacokinetic data in mice, rats, and cynomolgus monkeys were used to predict the pharmacokinetics of rhuMAb VEGF using allometric scaling in humans. The predicted serum clearance of rhuMAb VEGF in humans was 2.4 ml/day/kg and the terminal half-life was 12 days. Two hours after i.v. bolus administration of 125I-rhuMAb VEGF in rabbits, trichloroacetic acid-precipitable radioactivity was noted primarily in the plasma, with lesser amounts in highly perfused tissues such as kidneys, testes, spleen, heart, and lungs. At 48 h after dosing, trichloroacetic acid-precipitable radioactivity was noted in plasma with minimal distribution to testes, bladder, heart, lungs, and kidneys. Tissue distribution and pharmacokinetic data indicate that rhuMAb VEGF is cleared slowly and distributes to specific sites in the body. PMID- 9862793 TI - MCAT is not required for in vitro polyketide synthesis in a minimal actinorhodin polyketide synthase from Streptomyces coelicolor. AB - BACKGROUND: It has been proposed that Streptomyces malonyl CoA: holo acyl carrier protein transacylases (MCATs) provide a link between fatty acid and polyketide biosynthesis. Two recent studies have provided evidence that the presence of MCAT is essential for polyketide synthesis to proceed in reconstituted minimal polyketide synthases (PKSs). In contrast to this, we previously showed that the holo acyl carrier proteins (ACPs) from type II PKSs are capable of catalytic self malonylation in the presence of malonyl CoA, which suggests that MCAT might not be necessary for polyketide biosynthesis. RESULTS: We reconstituted a homologous actinorhodin (act) type II minimal PKS in vitro. When act holo-ACP is present in limiting concentrations, MCAT is required by the synthase complex in order for polyketide biosynthesis to proceed. When holo-ACP is present in excess, however, efficient polyketide synthesis proceeds without MCAT. The rate of polyketide production increases with holo-ACP concentration, but at low ACP concentration or equimolar AC:KS:CLF (KS, ketosynthase; CLF, chain length determining factor) concentrations this rate is significantly lower than expected, indicating that free holo-ACP is sequestered by the KS/CLF complex. CONCLUSIONS: The rate of polyketide biosynthesis is dictated by the ratio of holo-ACP to KS and CLF, as well as by the total protein concentration. There is no absolute requirement for MCAT in polyketide biosynthesis in vitro, although the role of MCAT during polyketide synthesis in vivo remains an open question. MCAT might be responsible for the rate enhancement of malonyl transfer at very low free holo-ACP concentrations or it could be required to catalyse the transfer of malonyl groups from malonyl CoA to sequestered holo-ACP. PMID- 9862794 TI - In vitro evolution of molecular cooperation in CATCH, a cooperatively coupled amplification system. AB - BACKGROUND: One of the key issues in the investigation of evolution is how complex systems evolved from simple chemical replicators. Theoretical work proposed several models in which complex replicating systems are kinetically stabilized. The development of powerful isothermal amplification technique allows complex nucleic acid based evolving in vitro systems to be set up, which may then serve to verify experimentally current theories of evolution. Recently such a system based on the 3SR (self-sustained sequence replication) reaction has been established to investigate the evolution of cooperation: the trans-cooperatively coupled CATCH (cooperative amplification by cross hybridization). RESULTS: Over four rounds of serial transfer, the cooperatively coupled two species CATCH system evolved into a more complex cooperative four species system, which then was overgrown by CATCH-derived RNA-Z-like hairpin species. In contrast to the classical RNA-Z species, these molecules have complementary loop sequences and self-amplify using a dual mechanism that includes concentration-dependent phases of noncooperative and cooperative amplification. CONCLUSIONS: The evolution of a cooperative system, under conditions that were alternately unfavorable and favorable for cooperative amplification, led to a system showing facultative cooperation. This principle of facultative cooperation preserves the complexity of the system investigated and could have general implications for the evolution and stabilization of cooperation under oscillating reaction conditions. PMID- 9862795 TI - Strategies for protein-based nanofabrication: Ni2+-NTA as a chemical mask to control biologically imposed symmetry. AB - BACKGROUND: Technologies that improve control of protein orientation on surfaces or in solution, through designed molecular recognition, will expand the range of proteins that are useful for biosensors, molecular devices and biomaterials. A limitation of some proteins is their biologically imposed symmetry, which results in indistinguishable recognition surfaces. Here, we have explored methods for modifying the symmetry of an oligomeric protein that exhibits useful self assembly properties. RESULTS: Escherichia coli glutamine synthetase (GS) contains 24 solvent-exposed histidines on two symmetry-related surfaces. These histidines drive a metal-dependent self-assembly of GS tubes. Immobilization of GS on the affinity resin Ni2+-NTA followed by on-column modification with diethyl pyrocarbonate affords asymmetrically modified GS that self-assembles only to the extent of 'short' dimeric GS tubes, as demonstrated by electron microscopy, dynamic light scattering and atomic force microscopy. The utility of Ni2+-NTA as a chemical mask was also demonstrated for asymmetric modification of engineered cysteines adjacent to the natural histidines. CONCLUSIONS: Current genetic methods do not provide distinguishable recognition elements on symmetry-related surfaces of biologically assembled proteins. Ni2+-NTA serves as a mask to control chemical modification in vitro of residues within symmetry-related pairs, on proteins containing functional His-tags. This strategy may be extended to modification of a wide range of amino acids with a myriad of reagents. PMID- 9862796 TI - Oxidation of guanines in the iron-responsive element RNA: similar structures from chemical modification and recent NMR studies. AB - BACKGROUND: The translation or stability of the mRNAs from ferritin, maconitase, erythroid aminoevulinate synthase and the transferrin receptor is controlled by the binding of two iron regulatory proteins to a family of hairpin-forming RNA sequences called iron-responsive elements (IREs). The determination of high resolution nuclear magnetic resonance (NMR) structures of IRE variants suggests an unusual hexaloop structure, leading to an intra-loop G-C base pair and a highly exposed loop guanine, and a special internal loop/bulge in the ferritin IRE involving a shift in base pairing not predicted with standard algorithms. RESULTS: Cleavage of synthetic 55- and 30-mer RNA oligonucleotides corresponding to the ferritin IRE with complexes based on oxoruthenium(IV) shows enhanced reactivity at a hexaloop guanine and at a guanine adjacent to the internal loop/bulge with strong protection at a guanine in the internal loop/bulge. These results are consistent with the recent NMR structures. The synthetic 55-mer RNA binds the iron-regulatory protein from rabbit reticulocyte lysates. The DNA analogs of the 55- and 30-mers do not show the same reactivity pattern. CONCLUSIONS: The chemical reactivity of the guanines in the ferritin IRE towards oxoruthenium(IV) supports the published NMR structures and the known oxidation chemistry of the metal complexes. The results constitute progress towards developing stand-alone chemical nucleases that reveal significant structural properties and provide results that can ultimately be used to constrain molecular modeling. PMID- 9862797 TI - New junction models for alternate-strand triple-helix formation. AB - BACKGROUND: [corrected] Oligonucleotide-directed triple-helix (triplex) formation can interfere with gene expression but only long tracts of oligopyrimidine*oligopurine sequences can be targeted. Attempts have been made to recognize short oligopurine sequences alternating on the two strands of double stranded DNA by the covalent linkage of two triplex-forming oligonucleotides. Here we focus on the rational optimization of such an alternate-strand triplex formation on a DNA duplex containing a 5'-GpT-3'/3'-CpA-5' or a 5'-TpG-3'/3'-ApC 5' step by combination of (G,T)- and (G,A)-containing oligonucleotides that bind to the oligopurine strands in opposite orientations. RESULTS: The deletion of one nucleotide in the reverse Hoogsteen region of the oligonucleotide provides the best binding at the 5'GpT-3'/3'-CpA-5' step, whereas the addition of two cytosines as a linker between the two oligonucleotides is the best strategy to cross a 5'-TpG-3'/3'-ApC-5' step. Energy minimization and experimental data suggest that these two cytosines are involved in the formation of two novel base quadruplets. CONCLUSIONS: These data provide a rational basis for the design of oligonucleotides capable of binding to oligopurine sequences that alternate on the two strands of double-stranded DNA with a 5'-GpT-3'/3'-CpA-5' or a 5'-TpG 3'/3'-ApC-5' step at the junction. PMID- 9862798 TI - A FLASH of insight into cellular chemistry: genetically encoded labels for protein visualization in vivo. AB - Genetically encoded fluorescent labels, such as green fluorescent protein, make it possible to visualize a protein's natural distribution and environment in living cells. A new approach to protein labeling in living cells has been devised in which a small, membrane-permeable ligand binds with high affinity and specificity to a short peptide motif that can be incorporated into the protein of interest; the ligand becomes brightly fluorescent after binding to the peptide. PMID- 9862799 TI - Evolution of peptides that modulate the spectral qualities of bound, small molecule fluorophores. AB - BACKGROUND: Fluorophore dyes are used extensively in biomedical research to sensitively assay cellular constituents and physiology. We have created, as proof of principle, fluorophore dye binding peptides that could have applications in fluorescent dye-based approaches in vitro and in vivo. RESULTS: A panel of Texas red, Rhodamine red, Oregon green 514 and fluorescein binding peptides, termed here 'fluorettes', was selected via biopanning of a combinatorial library of 12 mer peptides fused to a minor coat pIII protein of the filamentous bacteriophage M13. The 'best' fluorette sequences from each of the groups were subjected to further mutagenesis, followed by a second biopanning to select a new generation of improved fluorettes. Phage were selected that had higher avidity for each fluorophore except Rhodamine red. Of these, peptides were characterized that could specifically and with high affinity bind at least one dye, Texas red, in solution. In addition, the binding of certain peptides to Texas red shifted the peak excitation and/or the emission spectra of the bound dye. CONCLUSIONS: Peptides in the context of phage display could readily be selected that could bind to small-molecule fluorophores. The affinities of selected mutant fluorettes could be increased by mutation and further selection. Only a subset of the free peptides could bind free dyes in solution, suggesting that phage context contributed to the selection and ability of certain peptidic regions to independently bind the dyes. Future screens might lead to the creation of other dye-binding peptides with novel characteristics or Texas red derivatives with cross-linking substituents might be designed to increase the utility of the system. PMID- 9862800 TI - Evaluating precursor-directed biosynthesis towards novel erythromycins through in vitro studies on a bimodular polyketide synthase. AB - BACKGROUND: Modular polyketide synthases (PKSs) catalyse the biosynthesis of complex polyketides using a different set of enzymes for each successive cycle of chain extension. Directed biosynthesis starting from synthetic diketides is a potentially valuable route to novel polyketides. We have used a purified bimodular derivative of the erythromycin-producing polyketide synthase (DEBS 1 TE) to study chain extension starting from a variety of diketide analogues and, in some cases, from the alternative acyl-CoA thioester substrates. RESULTS: Chain initiation in vitro by DEBS 1-TE module 2 using a synthetic diketide analogue as a substrate was tolerant of significant structural variation in the starter unit of the synthetic diketide, but other changes completely abolished activity. Interestingly, a racemic beta-keto diketide was found to be reduced in situ on the PKS and utilised in place of its more complex hydroxy analogue as a substrate for chain extension. The presence of a diketide analogue strongly inhibited chain initiation via the loading module. Significantly higher concentrations of diketide N-acetylcysteamine analogues than their corresponding acyl-CoA thioesters are required to achieve comparable yields of triketide lactones. CONCLUSIONS: Although a broad range of variation in the starter residue is acceptable, the substrate specificity of module 2 of a typical modular PKS in vitro is relatively intolerant of changes at C-2 and C-3. This will restrict the usefulness of approaches to synthesise novel erythromycins using synthetic diketides in vivo. The use of synthetic beta-keto diketides in vivo deserves to be explored. PMID- 9862801 TI - Inner space exploration: the chemical biologist's guide to the cell. PMID- 9862802 TI - Radically novel prostaglandins in animals and plants: the isoprostanes. AB - Animal prostaglandins and plant jasmonates are well-known enzymatically formed cyclopentanoic lipids that have regulatory functions and serve as inducible mediators of host defense reactions. A novel group of prostaglandin-like compounds, the isoprostanes, generated in animals and plants by a nonenzymatic, free radical-catalyzed process, are now suspected to be mediators of oxidant injury in vivo. PMID- 9862803 TI - Crystal structure and conformation of a DNA-RNA hybrid duplex with a polypurine RNA strand: d(TTCTTBr5CTTC)-r(GAAGAAGAA). AB - BACKGROUND: . DNA-RNA hybrids are substrates for RNase H. This enzyme catalyzes the hydrolysis of the RNA strand in the hybrid form. The polypurine tract (PPT) in human immunodeficiency virus 1 (HIV-1) is a short stretch of purines ( approximately 15 bases) located at the 3'-end of the U3 region of the RNA genome. The PPT has the unique ability to resist digestion by RNase H and serves as a primer for plus-strand DNA synthesis. RESULTS: . The crystal structure of a DNA RNA hybrid duplex containing a polypurine RNA strand, d(TTCTTBr5CTTC) r(GAAGAAGAA), has been determined at 1.8 A resolution. The structure was solved by molecular replacement methods and refined to a final R factor of 20.1% (R free 23.7%). The hybrid duplex adopts a standard A-form conformation. All of the sugar rings and glycosidic torsion angles are found in the standard C3'-endo/anti conformation, as seen in A-RNA or A-DNA. The crystal packing is dominated by the DNA strand, where the terminal base pairs of the hybrid abut the neighboring A DNA sugar-phosphate backbone on the minor groove side. CONCLUSIONS: . The present DNA-RNA hybrid duplex containing a polypurine RNA strand exhibits standard A-form geometry. This observation might suggest that the RNA PPT resists the RNase H activity of HIV reverse transcriptase as a result of its A-form conformation. In addition, there appears to be a correlation between the percentage purine content of the RNA and the DNA backbone conformation. PMID- 9862804 TI - Structures of the psychrophilic Alteromonas haloplanctis alpha-amylase give insights into cold adaptation at a molecular level. AB - BACKGROUND: . Enzymes from psychrophilic (cold-adapted) microorganisms operate at temperatures close to 0 degreesC, where the activity of their mesophilic and thermophilic counterparts is drastically reduced. It has generally been assumed that thermophily is associated with rigid proteins, whereas psychrophilic enzymes have a tendency to be more flexible. RESULTS: . Insights into the cold adaptation of proteins are gained on the basis of a psychrophilic protein's molecular structure. To this end, we have determined the structure of the recombinant form of a psychrophilic alpha-amylase from Alteromonas haloplanctis at 2.4 A resolution. We have compared this with the structure of the wild-type enzyme, recently solved at 2.0 A resolution, and with available structures of their mesophilic counterparts. These comparative studies have enabled us to identify possible determinants of cold adaptation. CONCLUSIONS: . We propose that an increased resilience of the molecular surface and a less rigid protein core, with less interdomain interactions, are determining factors of the conformational flexibility that allows efficient enzyme catalysis in cold environments. PMID- 9862805 TI - Structures of escherichia coli CMP kinase alone and in complex with CDP: a new fold of the nucleoside monophosphate binding domain and insights into cytosine nucleotide specificity. AB - BACKGROUND: . Nucleoside monophosphate kinases (NMP kinases) catalyze the reversible transfer of a phosphoryl group from a nucleoside triphosphate to a nucleoside monophosphate. Among them, cytidine monophosphate kinase from Escherichia coli has a striking particularity: it is specific for CMP, whereas in eukaryotes a unique UMP/CMP kinase phosphorylates both CMP and UMP with similar efficiency. RESULTS: . The crystal structure of the CMP kinase apoenzyme from E. coli was solved by single isomorphous replacement and refined at 1.75 A resolution. The structure of the enzyme in complex with CDP was determined at 2.0 A resolution. Like other NMP kinases, the protein contains a central parallel beta sheet, the strands of which are connected by alpha helices. The enzyme differs from other NMP kinases in the presence of a 40-residue insert situated in the NMP-binding (NMPbind) domain. This insert contains two domains: one comprising a three-stranded antiparallel beta sheet, the other comprising two alpha helices. CONCLUSIONS: . Two features of the CMP kinase from E. coli have no equivalent in other NMP kinases of known structure. Firstly, the large NMPbind insert undergoes a CDP-induced rearrangement: its beta-sheet domain moves away from the substrate, whereas its helical domain comes closer to it in a motion likely to improve the protection of the active site. Secondly, residues involved in CDP recognition are conserved in CMP kinases and have no counterpart in other NMP kinases. The structures presented here are the first of a new family of NMP kinases specific for CMP. PMID- 9862806 TI - Ligand size is a major determinant of specificity in periplasmic oxyanion-binding proteins: the 1.2 A resolution crystal structure of Azotobacter vinelandii ModA. AB - BACKGROUND: . Periplasmic receptors constitute a diverse class of binding proteins that differ widely in size, sequence and ligand specificity. Nevertheless, almost all of them display a common beta/alpha folding motif and have similar tertiary structures consisting of two globular domains. The ligand is bound at the bottom of a deep cleft, which lies at the interface between these two domains. The oxyanion-binding proteins are notable in that they can discriminate between very similar ligands. RESULTS: . Azotobacter vinelandii is unusual in that it possesses two periplasmic molybdate-binding proteins. The crystal structure of one of these with bound ligand has been determined at 1.2 A resolution. It superficially resembles the structure of sulphate-binding protein (SBP) from Salmonella typhimurium and uses a similar constellation of hydrogen bonding interactions to bind its ligand. However, the detailed interactions are distinct from those of SBP and the more closely related molybdate-binding protein of Escherichia coli. CONCLUSIONS: . Despite differences in the residues involved in binding, the volumes of the binding pockets in the A. vinelandii and E. coli molybdate-binding proteins are similar and are significantly larger than that of SBP. We conclude that the discrimination between molybdate and sulphate shown by these binding proteins is largely dependent upon small differences in the sizes of these two oxyanions. PMID- 9862807 TI - Sheep liver cytosolic aldehyde dehydrogenase: the structure reveals the basis for the retinal specificity of class 1 aldehyde dehydrogenases. AB - BACKGROUND: . Enzymes of the aldehyde dehydrogenase family are required for the clearance of potentially toxic aldehydes, and are essential for the production of key metabolic regulators. The cytosolic, or class 1, aldehyde dehydrogenase (ALDH1) of higher vertebrates has an enhanced specificity for all-trans retinal, oxidising it to the powerful differentiation factor all-trans retinoic acid. Thus, ALDH1 is very likely to have a key role in vertebrate development. RESULTS: . The three-dimensional structure of sheep ALDH1 has been determined by X-ray crystallography to 2.35 A resolution. The overall tertiary and quaternary structures are very similar to those of bovine mitochondrial ALDH (ALDH2), but there are important differences in the entrance tunnel for the substrate. In the ALDH1 structure, the sidechain of the general base Glu268 is disordered and the NAD+ cofactor binds in two distinct modes. CONCLUSIONS: . The submicromolar Km of ALDH1 for all-trans retinal, and its 600-fold enhanced affinity for retinal compared to acetaldehyde, are explained by the size and shape of the substrate entrance tunnel in ALDH1. All-trans retinal fits into the active-site pocket of ALDH1, but not into the pocket of ALDH2. Two helices and one surface loop that line the tunnel are likely to have a key role in defining substrate specificity in the wider ALDH family. The relative sizes of the tunnels also suggest why the bulky alcohol aversive drug disulfiram reacts more rapidly with ALDH1 than ALDH2. The disorder of Glu268 and the observation that NAD+ binds in two distinct modes indicate that flexibility is a key facet of the enzyme reaction mechanism. PMID- 9862808 TI - The crystal structure of pneumococcal surface antigen PsaA reveals a metal binding site and a novel structure for a putative ABC-type binding protein. AB - BACKGROUND: . The surface protein PsaA of the pathogenic bacterium Streptococcus pneumoniae plays an essential role in its virulence. PsaA is a putative ATP binding cassette-type (ABC-type) binding protein involved in the uptake of Mn2+ and possibly Zn2+ and is considered to be both a potential drug target and and a candidate vaccine component. RESULTS: . The structure of PsaA has been determined to 2.0 A resolution using X-ray crystallography and is the first structure obtained for an ABC-type binding protein from a Gram-positive organism. The protein consists of two (beta/alpha)4 domains linked together by a single helix. A metal-binding site is formed in the domain interface by the sidechains of His67, His139, Glu205 and Asp280 and is occupied in the structure. CONCLUSIONS: . The structural topology of PsaA is fundamentally different from that of other ABC type binding proteins determined thus far in that PsaA lacks the characteristic 'hinge peptides' involved in conformational change upon solute uptake and release. In our structure, the metal-binding site is probably occupied by Zn2+. The site seems to be well conserved amongst related receptors from both Gram positive and Gram-negative bacteria. PMID- 9862809 TI - Crystal structure of the DpnM DNA adenine methyltransferase from the DpnII restriction system of streptococcus pneumoniae bound to S-adenosylmethionine. AB - BACKGROUND: . Methyltransferases (Mtases) catalyze the transfer of methyl groups from S-adenosylmethionine (AdoMet) to a variety of small molecular and macromolecular substrates. These enzymes contain a characteristic alpha/beta structural fold. Four groups of DNA Mtases have been defined and representative structures have been determined for three groups. DpnM is a DNA Mtase that acts on adenine N6 in the sequence GATC; the enzyme represents group alpha DNA Mtases, for which no structures are known. RESULTS: . The structure of DpnM in complex with AdoMet was determined at 1.80 A resolution. The protein comprises a consensus Mtase fold with a helical cluster insert. DpnM binds AdoMet in a similar manner to most other Mtases and the enzyme contains a hollow that can accommodate DNA. The helical cluster supports a shelf within the hollow that may recognize the target sequence. Modeling studies indicate a potential site for binding the target adenine, everted from the DNA helix. Comparison of the DpnM structure and sequences of group alpha DNA Mtases indicates that the group is a genetically related family. Structural comparisons show DpnM to be most similar to a small-molecule Mtase and then to macromolecular Mtases, although several dehydrogenases show greater similarity than one DNA Mtase. CONCLUSIONS: . DpnM, and by extension the DpnM family or group alpha Mtases, contains the consensus fold and AdoMet-binding motifs found in most Mtases. Structural considerations suggest that macromolecular Mtases evolved from small-molecule Mtases, with different groups of DNA Mtases evolving independently. Mtases may have evolved from dehydrogenases. Comparison of these enzymes indicates that in protein evolution, the structural fold is most highly conserved, then function and lastly sequence. PMID- 9862810 TI - The crystal structure of ribosomal protein L22 from Thermus thermophilus: insights into the mechanism of erythromycin resistance. AB - BACKGROUND: . The ribosomal protein L22 is one of five proteins necessary for the formation of an early folding intermediate of the 23S rRNA. L22 has been found on the cytoplasmic side of the 50S ribosomal subunit. It can also be labeled by an erythromycin derivative bound close to the peptidyl-transfer center at the interface side of the 50S subunit, and the amino acid sequence of an erythromycin resistant mutant is known. Knowing the structure of the protein may resolve this apparent conflict regarding the location of L22 on the ribosome. RESULTS: . The structure of Thermus thermophilus L22 was solved using X-ray crystallography. L22 consists of a small alpha+beta domain and a protruding beta hairpin that is 30 A long. A large part of the surface area of the protein has the potential to be involved in interactions with rRNA. A structural similarity to other RNA-binding proteins is found, possibly indicating a common evolutionary origin. CONCLUSIONS: . The extensive surface area of L22 has the characteristics of an RNA-binding protein, consistent with its role in the folding of the 23S rRNA. The erythromycin-resistance conferring mutation is located in the protruding beta hairpin that is postulated to be important in L22-rRNA interactions. This region of the protein might be at the erythromycin-binding site close to the peptidyl transferase center, whereas the opposite end may be exposed to the cytoplasm. PMID- 9862811 TI - Crystal structure of quinolinic acid phosphoribosyltransferase from Mmycobacterium tuberculosis: a potential TB drug target. AB - BACKGROUND: . Mycobacterium tuberculosis is the single most deadly human pathogen and is responsible for nearly three million deaths every year. Recent elucidation of the mode of action of isoniazid, a frontline antimycobacterial drug, suggests that NAD metabolism is extremely critical for this microorganism. M. tuberculosis depends solely on the de novo pathway to meet its NAD demand. Quinolinic acid phosphoribosyltransferase (QAPRTase), a key enzyme in the de novo biosynthesis of NAD, provides an attractive target for designing novel antitubercular drugs. RESULTS: . The X-ray crystal structure of the M. tuberculosis QAPRTase apoenzyme has been determined by multiple isomorphous replacement at 2.4 A resolution. Structures of the enzyme have also been solved in complex with the substrate quinolinic acid (QA), the inhibitory QA analog phthalic acid (PA), the product nicotinate mononucleotide (NAMN), and as a ternary complex with PA and a substrate analog, 5-phosphoribosyl-1-(beta-methylene)pyrophosphate (PRPCP). The structure of the nonproductive QAPRTase-PA-PRPCP Michaelis complex reveals a 5 phosphoribosyl-1-pyrophosphate-binding site that is different from the one observed in type I phosphoribosyltransferases (PRTases). The type II PRTase active site of QAPRTase undergoes conformational changes that appear to be important in determining substrate specificity and eliciting productive catalysis. CONCLUSIONS: . QAPRTase is the only known representative of the type II PRTase fold, an unusual alpha/beta barrel, and appears to represent convergent evolution for PRTase catalysis. The active site of type II PRTase bears little resemblance to the better known type I enzymes. PMID- 9862812 TI - GDP-fucose synthetase from Escherichia coli: structure of a unique member of the short-chain dehydrogenase/reductase family that catalyzes two distinct reactions at the same active site. AB - BACKGROUND: . In all species examined, GDP-fucose is synthesized from GDP-mannose in a three-step reaction catalyzed by two enzymes, GDP-mannose 4,6 dehydratase and a dual function 3, 5-epimerase-4-reductase named GDP-fucose synthetase. In this latter aspect fucose biosynthesis differs from that of other deoxy and dideoxy sugars, in which the epimerase and reductase activities are present as separate enzymes. Defects in GDP-fucose biosynthesis have been shown to affect nodulation in bacteria, stem development in plants, and are associated with the immune defect leukocyte adhesion deficiency type II in humans. RESULTS: . We have determined the structure of GDP-fucose synthetase from Escherichia coli at 2.2 A resolution. The structure of GDP-fucose synthetase is closely related to that of UDP-galactose 4-epimerase and more distantly to other members of the short-chain dehydrogenase/reductase family. We have also determined the structures of the binary complexes of GDP-fucose synthetase with its substrate NADPH and its product NADP+. The nicotinamide cofactors bind in the syn and anti conformations, respectively. CONCLUSIONS: . GDP-fucose synthetase binds its substrate, NADPH, in the proper orientation (syn) for transferring the 4-pro-S hydride of the nicotinamide. We have observed a single binding site in GDP-fucose synthetase for the second substrate, GDP-4-keto,6-deoxy-mannose. This implies that both the epimerization and reduction reactions occur at the same site in the enzyme. As is the case for all members of the short-chain family of dehydrogenase/reductases, GDP-fucose synthetase retains the Ser-Tyr-Lys catalytic triad. We propose that this catalytic triad functions in a mechanistically equivalent manner in both the epimerization and reduction reactions. Additionally, the X-ray structure has allowed us to identify other residues that are potentially required for substrate binding and catalysis. PMID- 9862813 TI - Springs and zippers: coiled coils in SNARE-mediated membrane fusion. AB - A conserved molecular machinery based on SNARE proteins catalyzes most, if not all, cellular membrane fusion events. A flurry of recent biophysical studies have established a detailed molecular picture of the core SNARE complex. Structural and biochemical analysis of the SNARE machinery is rapidly advancing our understanding of the specificity, regulation and protein catalysis of membrane fusion. PMID- 9862815 TI - Intestinal IgA: novel views on its function in the defence of the largest mucosal surface. PMID- 9862816 TI - Transabdominal bowel sonography in Crohn's disease. PMID- 9862817 TI - Nitric oxide and the migrating motor complex. PMID- 9862818 TI - Chronic pancreatitis and mutations of the cystic fibrosis gene. PMID- 9862819 TI - Ranitidine reduced clinically important gastrointestinal bleeding in patients who required mechanical ventilation. PMID- 9862820 TI - Dietary polyamines are essential luminal growth factors for small intestinal and colonic mucosal growth and development. AB - BACKGROUND: Polyamines are essential for cell growth. Dietary and probably gut bacterial derived polyamines contribute significantly to the polyamine body pool. AIMS: To evaluate the influence of dietary, luminal polyamines on growth and development of different gastrointestinal organs in normally growing rats. METHODS: Male suckling Wistar rats were randomly allocated to four treatment groups: polyamine deficient diet (PDD); PDD plus antibiotics (neomycin 2 g/kg and metronidazole 34 mg/kg); PDD plus polyamine supplementation at normal concentrations; or normal standard laboratory chow. After a six month feeding period 7-10 animals/group were sacrificed. RESULTS: No differences in body weight gain, food consumption, or general behaviour could be observed between the four groups of animals. Feeding of PDD alone or PDD plus antibiotics resulted in a highly significant decrease in organ weight, protein content, and DNA content in small intestinal and colonic mucosa whereas no alterations were found in the liver. CONCLUSIONS: Long term feeding of polyamine deficient diets resulted in a significant hypoplasia of small intestinal and colonic mucosa. Dietary, luminal polyamines are important local factors for growth and the development of small intestinal and colonic mucosa. PMID- 9862821 TI - Increased expression of mRNA for matrix metalloproteinases-1 and -3 and tissue inhibitor of metalloproteinases-1 in intestinal biopsy specimens from patients with coeliac disease. AB - BACKGROUND: Extracellular matrix (ECM) degradation may play a role in villus atrophy in coeliac disease (CD). AIMS: To compare the cellular expression of mRNA transcripts for the two major matrix degrading proteases, matrix metalloproteinase (MMP)-1 and MMP-3, their inhibitor, tissue inhibitor of metalloproteinases (TIMP)-1, and procollagen I in the intestinal mucosa of patients with untreated and treated CD and normal controls. PATIENTS/METHODS: Duodenal biopsy specimens from ten untreated CD patients, from six of these after a gluten free diet, and from ten control patients were hybridised with 35S labelled RNA probes. The number of positive cells in the subepithelial region and lamina propria were counted microscopically. RESULTS: The numbers of cells positive for MMP-1 (p<0.005), MMP-3 (p<0.01), and TIMP-1 (p<0.05) mRNA were higher in the subepithelial region of CD mucosa than in that from controls. In the lamina propria, only cells positive for MMP-1 mRNA were increased in CD patients compared with controls (p<0.01). MMP-1 and MMP-3 mRNA expression returned to normal in CD patients after treatment with a gluten free diet (p<0.05), while TIMP-1 mRNA expression remained elevated. The number of procollagen I mRNA expressing cells did not change. Expression of MMP-1 and MMP-3 mRNA was mainly localised to subepithelial fibroblasts and macrophages. CONCLUSIONS: The decreased ratio of collagen I and TIMP-1 mRNA expressing cells to MMP-1 and MMP-3 mRNA expressing cells in untreated CD suggests a shift towards ECM degradation. ECM degradation by activated subepithelial fibroblasts and macrophages may be an important mechanism driving mucosal transformation in CD. PMID- 9862822 TI - The effect of epidermal growth factor on brush border surface area and function in the distal remnant following resection in the rabbit. AB - BACKGROUND: Epidermal growth factor (EGF) has been shown to increase intestinal absorptive surface area and transport function in normal animals. AIMS: To examine the effect of EGF on absorptive surface area and brush border membrane function in a model of massive small bowel resection. METHODS: New Zealand white rabbits were randomised into two groups: a resected group (60% proximal small bowel resection); and an unmanipulated control group. Distal remnant tissue was examined 10 and 21 days postsurgery. In separate experiments oral EGF (40 g/kg/day) was administered to resected animals from days 3 to 8 and animals were studied on day 10. RESULTS: Ten days postsurgery brush border surface area and total absorptive surface area were significantly increased in remnant tissue while brush border membrane vesicle (BBMV) glucose uptake was significantly decreased compared with controls. By 21 days brush border surface area returned to control levels though BBMV glucose uptake remained depressed. EGF treatment induced a further increase in brush border surface area in remnant intestine but did not alter BBMV glucose uptake. CONCLUSIONS: Surgical resection results in significant elevations in absorptive surface area coupled with a decrease in brush border membrane transport function distal to the site of anastomosis. EGF enhances glucose uptake in remnant intestine via recruitment of additional microvillus membrane into the brush border. PMID- 9862824 TI - Potassium secretion in rat distal colon during dietary potassium loading: role of pH regulated apical potassium channels. AB - BACKGROUND: Chronic dietary K+ loading increases the abundance of large conductance (210 pS) apical K+ channels in surface cells of rat distal colon, resulting in enhanced K+ secretion in this epithelium. However, the factors involved in the regulation of these K+ channels are at present unclear. AIMS: To evaluate the effect of dietary K+ loading on intracellular pH and its relation to large conductance apical K+ channel activity in surface cells of rat distal colon. METHODS/RESULTS: As assessed by fluorescent imaging, intracellular pH was higher in K+ loaded animals (7.48 (0.09)) than in controls (7.07 (0.04); p<0.01) when surface cells were bathed in NaCl solution, and a similar difference in intracellular pH was observed when cells were bathed in Na2SO4 solution (7.67 (0.09) and 6.92 (0.05) respectively; p<0.001). Ethylisopropylamiloride (EIPA; an inhibitor of Na+-H+ exchange; 1 microM) decreased intracellular pH when surface cells from K+ loaded animals were bathed in either solution, although the decrease was greater when the solution contained NaCl (DeltapH 0.50 (0.03)) rather than Na2SO4 (DeltapH 0. 18 (0.02); p<0.05). In contrast, EIPA had no effect in cells from control animals. As assessed by patch clamp recording techniques, the activity of large conductance K+ channels in excised inside-out membrane patches from distal colonic surface cells of K+ loaded animals increased twofold when the bath pH was raised from 7.40 to 7. 60. As assessed by cell attached patches in distal colonic surface cells from K+ loaded animals, the addition of 1 M EIPA decreased K+ channel activity by 50%, consistent with reversal of Na+-H+ exchange mediated intracellular alkalinisation. CONCLUSION: Intracellular alkalinisation stimulates pH sensitive large conductance apical K+ channels in rat distal colonic surface cells as part of the K+ secretory response to chronic dietary K+ loading. Intracellular alkalinisation seems to reflect an increase in EIPA sensitive Na+-H+ exchange, which may be a manifestation of the secondary hyperaldosteronism associated with this model of colonic K+ adaptation. PMID- 9862823 TI - Proabsorptive and prosecretory roles for nitric oxide in cholera toxin induced secretion. AB - BACKGROUND: Cholera toxin causes small intestinal hypersecretion by inducing a coordinated response from enterocytes, enterochromaffin cells, enteric neurones, and the vascular supply. Nitric oxide has been implicated in the function of these separate components. AIMS: To explore the role of nitric oxide in the totality of cholera toxin induced secretion in vivo. METHODS: One group of adult male Wistar rats was treated with the nitric oxide synthase inhibitors NG-nitro-L arginine methyl ester (L-NAME; subcutaneously or intraluminally), NG-methyl-L arginine (L-NMA), or 7-nitroindazole. A second group of rats was treated with L arginine (intraperitoneally or intraluminally) or D-arginine. The small intestine was isolated between two cannulae and instilled with 75 microg cholera toxin or saline for two hours. Small intestinal perfusion of a plasma electrolyte solution containing [14C]-PEG was undertaken to determine net water and electrolyte movement. After the experiment macroscopic and microscopic intestinal appearances were noted and jejunal 5-hydroxytryptamine concentrations were determined. RESULTS: Both L-arginine and L-NAME induced secretion in the basal state, but only when administered intraluminally. Systemically applied L-NAME caused a dose dependent reduction in cholera toxin induced secretion. This was paralleled by L NMA but not by 7-nitroindazole or by intraluminally applied L-NAME. Systemically applied L-NAME caused notable cyanosis of the intestine, consistent with mesenteric ischaemia, but no microscopic abnormalities. Systemically applied L arginine but not D-arginine also reduced cholera toxin induced secretion and inhibited 5-hydroxytryptamine release. CONCLUSION: Nitric oxide has a duality of roles in cholera toxin induced secretion, acting both as an absorbagogue and a secretagogue. Its mechanisms of action include the maintenance of mucosal perfusion and enterochromaffin cell stabilisation. PMID- 9862825 TI - Patterns of gas and liquid reflux during transient lower oesophageal sphincter relaxation: a study using intraluminal electrical impedance. AB - BACKGROUND: Belching has been proposed as a major mechanism underlying acid gastro-oesophageal reflux in normal subjects. However, the presence of oesophageal gas has not been measured directly but only inferred from manometry. AIMS: To investigate, using intraluminal electrical impedance, the patterns of gas and liquid reflux during transient lower oesophageal sphincter (LOS) relaxations, the main mechanism of acid reflux in normal subjects. METHODS: Impedance changes associated with the passage of gas were studied in vitro, and in vivo in cats. Oesophageal manometry, pH, and intraluminal electrical impedance measurements were performed in 11 normal subjects after a meal. RESULTS: Gas reflux caused a sudden increase in impedance that propagated rapidly to the proximal oesophagus whereas liquid reflux induced a retrogressively propagated fall in impedance. Impedance showed gas or liquid reflux during most (102/141) transient LOS relaxations. When acid reflux occurred, impedance showed evidence of intraoesophageal retrograde flow of liquid in the majority (78%) of events. Evidence of gas retroflow was found in almost half (47%) of acid reflux episodes. When present together, however, liquid preceded gas on 44% of occasions. Overall, gas reflux occurred as the initial event in only 25% of acid reflux episodes. CONCLUSIONS: These findings suggest that in upright normal subjects, although belching can precipitate acid reflux, most acid reflux occurs as a primary event. PMID- 9862826 TI - Gastric accommodation in non-ulcer dyspepsia and the roles of Helicobacter pylori infection and vagal function. AB - BACKGROUND: The pathophysiological mechanisms in non-ulcer dyspepsia are incompletely understood. AIMS: To compare gastric motor and sensory functions in Helicobacter pylori positive or negative patients with non-ulcer dyspepsia. PATIENTS: Seventeen patients with non-ulcer dyspepsia and 16 asymptomatic controls. METHODS: The following were evaluated: gastrointestinal symptoms; gastric emptying and orocaecal transit of solids; abdominal vagal function; gastric compliance; fasting and postprandial gastric tone and phasic contractions; symptoms during ingestion of cold water and during the distension of an intragastric bag; and somatic sensitivity and personality profile (Minnesota Multiphasic Personality Inventory, MMPI). RESULTS: Gastric accommodation was reduced in H pylori negative dyspeptics relative to controls; the degree of accommodation was unrelated to H pylori status in dyspeptics. Increased postprandial gastric sensation was more frequent among H pylori positive patients (4/5 H pylori positive versus 4/12 H pylori negative patients). Intragastric meal distribution and orocaecal transit were normal; gastric emptying at four hours was abnormal in 4/17 patients. Vagal dysfunction was rare. Eight of 17 patients had somatisation or depression on MMPI. CONCLUSION: Impaired gastric accommodation is frequent in non-ulcer dyspepsia and seems to be unrelated to vagal efferent dysfunction. H pylori infection does not seem to influence gastric accommodation, but is associated with heightened sensitivity in dyspeptics. Therapeutic approaches that restore normal postprandial accommodation and gastric sensitivity should be tested in non-ulcer dyspepsia. PMID- 9862827 TI - Interstitial cells of Cajal in the human fetal small bowel as shown by c-kit immunohistochemistry. AB - BACKGROUND: Interstitial cells of Cajal (ICCs) express the tyrosine kinase receptor c-kit, which is required for their development and spontaneous pacemaker activity in the bowel. From murine models it has been proposed that ICCs do not develop until after birth, but more recent findings indicate that c-kit is expressed early in the embryonic period. The temporal development of ICCs in the human gut remains unknown. AIM: To investigate ICCs in the human fetal small bowel using c-kit immunohistochemistry. SUBJECTS: Small bowel specimens were obtained at post mortem examination of 16 fetuses and nine neonates, eight of whom were premature, born at gestational ages of 13 to 41 weeks, without gastrointestinal disorders. METHODS: Immunohistochemical analysis was performed on material fixed in formalin and embedded in paraffin. The specimens were exposed to antibodies raised against c-kit (an ICC marker) and neurone specific enolase (a general neuronal marker). The ABC complex method was used to visualise binding of antibodies to the corresponding antigens. RESULTS: c-kit immunoreactive cells were visualised from 13 weeks of gestation. The immunoreactivity was mainly localised in association with the myenteric plexus. From about 17-18 weeks of gestation, the ICCs formed a layer along the myenteric plexus, whereas this layer appeared to be disrupted at 13-16 weeks of gestation. CONCLUSIONS ICCs are c-kit immunoreactive at least from a gestational age of 13 weeks in the human fetal small intestine. From 17-18 weeks of gestation until birth, they form a continuous layer around the myenteric ganglia. PMID- 9862828 TI - Evidence that nitric oxide mechanisms regulate small intestinal motility in humans. AB - BACKGROUND: Non-cholinergic non-adrenergic neural mechanisms involving nerves containing NO have been shown to modulate smooth muscle in the gastrointestinal tract, and it has been suggested that release from tonic NO inhibition may be important in the regulation of cyclical fasting small intestinal motility. AIMS: To evaluate the role of NO mechanisms in the regulation of fasting small intestinal motor activity in humans using a specific NO synthase inhibitor, NG monomethyl-L-arginine ( L-NMMA). METHODS: In seven healthy male volunteers, duodenal and jejunal pressures were measured for four hours with a nine lumen manometric catheter. Volunteers attended on four separate days on which they received an intravenous infusion either saline or L-NMMA (0.5, 2, or 4 mg/kg/h) in random order. Intravenous infusions began 10 minutes after completion of phase III of the migrating motor complex (MMC). RESULTS: The first episode of phase III activity occurred earlier after infusion of 2 and 4 mg/kg/h L-NMMA than after infusion of 0.5 mg/kg/h L-NMMA or saline (mean (95% confidence interval) 52 (36 68) and 57 (18-97) v 116 (69-193) and 145 (64-226) minutes respectively) with a resultant MMC cycle length of 82 (59-105) and 86 (46-126) v 132 (49-198) and 169 (98-240) minutes respectively. The total number of phase III activities during the four hour recording was increased (p<0.05) by L-NMMA at a dose of 4 mg/kg/h (2 (1-3)) but not at 2 mg/kg/h (1.5 (1-2)) or 0.5 mg/kg/h (1.3 (1-2)) compared with saline (1.3 (0.6-2)). L-NMMA had no effect on the duration, velocity, number of contractions per minute, length of migration, or site of origin of phase III of the MMC. The duration of phase I activity was shorter (p<0.05) with 4 mg/kg/h L-NMMA than with saline (12 (1-23) v 31 (19-44) minutes). CONCLUSIONS: These observations suggest that NO mechanisms play a role in the regulation of fasting small intestinal motor activity in humans. PMID- 9862829 TI - Prospective comparison of faecal incontinence grading systems. AB - BACKGROUND: Existing scales for assessing faecal incontinence have not been validated against clinical assessment, or with regard to reproducibility. They also fail to take into account faecal urgency, and the use of antidiarrhoeal medications. AIMS: To establish the validity, and sensitivity to change, of existing scales and a newly designed incontinence scale. METHODS: (1) Twenty three patients (21 females, median age 57 years) were prospectively evaluated by two independent clinical observers, using three established scales (Pescatori, Wexner, American Medical Systems), a newly devised scale which also includes details about urgency and antidiarrhoeal drugs, and by a 28 day diary. (2) A further 10 female patients were assessed by the same scales before and after surgery for faecal incontinence. RESULTS: (1) Assessments by two independent clinicians correlated well. All four scales and a diary card correlated highly and significantly with the clinical impression, with the new scale reaching the highest correlation (r=0.79, p<0.001). (2) All except one score changed significantly in response to surgical treatment; the new scale showed the greatest change, at the highest level of significance (p=0.004), and correlated best with the clinicians' assessment of change (r=0.94, p<0.001). CONCLUSIONS: Existing scales for the assessment of faecal incontinence correlate well with careful clinical impression of severity, and serve as useful and reproducible measures for comparison of patients and treatments. A newly devised scale has shown high clinical validity and utility. PMID- 9862830 TI - Glucagon-like peptide-1: a potent regulator of food intake in humans. AB - BACKGROUND/AIMS: Studies in animals suggest a physiological role for glucagon like peptide-1-(7-36)-amide (GLP-1) in regulating satiety. The role of GLP-1 in regulating food intake in man has, however, not been investigated. Subjects Sixteen healthy male subjects were examined in a double blind placebo controlled fashion. METHODS: The effect of graded intravenous doses (0, 0.375, 0.75, and 1.5 pmol/kg/min) of synthetic human GLP-1 on food intake and feelings of hunger and satiety was tested in healthy volunteers. RESULTS: Graded GLP-1 infusions resulted in a dose dependent reduction in food intake (maximal inhibition 35%, p<0.001 v control) and a similar reduction in calorie intake (32%; p<0.001). Fluid ingestion was also reduced by GLP-1 (18% reduction, p<0.01). No overt side effects were produced by GLP-1, but subjects experienced less hunger and early fullness in the period before a meal during GLP-1 infusion at the highest dose (p<0.05). CONCLUSIONS: Intravenous infusions of GLP-1 decrease spontaneous food intake even at physiological plasma concentrations, implying an important role for GLP-1 in the regulation of the early satiety response in humans. PMID- 9862831 TI - Percutaneous endoscopic gastrostomy feeding in patients with cystic fibrosis. AB - BACKGROUND: Malnutrition is a common management problem in patients with cystic fibrosis (CF). Various approaches to supplemental nutrition by both parenteral and enteral routes have been used. AIM: To analyse the efficacy and acceptability of supplemental overnight feeding using a percutaneous endoscopic gastrostomy (PEG) in patients with CF. PATIENTS: 53 patients with CF (43 adults; age >17 years) with severe pulmonary disease. METHODS: The technical success and complications of PEG insertion were documented together with changes in nutritional and pulmonary status of the cohort. RESULTS: PEG tubes were successfully inserted in all patients, with immediate complications (respiratory depression) in two (4%) and late complications in 13 (25%). Feeding was well tolerated by 50/51 (98%) of the cohort during a mean (SEM) follow up of 14.5 (2.1) months. The adult cohort had a significant increase in weight and body mass index at six months which was maintained at 12 months. Serum albumin concentration remained stable at six months but had fallen by 12 months, although the differences were not statistically significant. These results were reflected in the paediatric cohort. Pulmonary function in those followed up for one year had apparently stabilised, but the number of admissions to hospital over the year before and the year after PEG did not change. Half of the cohort were accepted for heart-lung/lung transplantation, the improvement in nutritional status being a prerequisite for this. CONCLUSION: Supplemental PEG tube feeding is well tolerated and results in a significant improvement in nutritional status and an apparent stabilisation of pulmonary function in severely malnourished CF patients with advanced pulmonary disease PMID- 9862832 TI - Crohn's disease severity in familial and sporadic cases. AB - BACKGROUND: Having a relative with inflammatory bowel disease increases the risk for Crohn's disease but may also increase its severity in affected patients. AIMS: To evaluate the influence of a family history on Crohn's disease course and severity. METHODS: 1316 patients followed in the same unit were studied retrospectively. Age at onset, duration of illness, site, and extent of disease were determined in patients with and without a family history. Additionally, disease severity was estimated by the need for medical therapy (steroid and immunosuppressive requirement) and the frequency and extent of excisional surgery. RESULTS: 152 (12%) patients had a family history of inflammatory bowel disease. Duration of follow up was longer in patients with a family history and there were more operations for perforating complications in familial cases. However, the importance of medical therapy, and the incidence and extent of excisional surgery were similar in familial and and sporadic cases. Kaplan-Meier estimated time to prescription of immunosuppressive drugs and first intestinal resection were similar in familial and sporadic cases. When the 152 patients with familial Crohn's disease were paired for sex, location of disease at onset, date of birth, and date of diagnosis with 152 patients sporadic Crohn's disease, the disease severity remained similar in the two groups of paired patients. CONCLUSION: Patients with Crohn's disease and a family history of inflammatory bowel disease do not have a more severe course. PMID- 9862833 TI - Different intestinal permeability patterns in relatives and spouses of patients with Crohn's disease: an inherited defect in mucosal defence? AB - BACKGROUND: A familial defect in intestinal barrier function has been found in Crohn's disease. AIM: To investigate possible genetic and environmental influences on this barrier defect by studying intestinal permeability in both relatives and spouses of patients with Crohn's disease. SUBJECTS: The study included 39 patients with Crohn's disease, 34 healthy first degree relatives, and 22 spouses. Twenty nine healthy volunteers served as controls. METHODS: Intestinal permeability was assessed as the lactulose:mannitol ratio in five hour urinary excretion after oral load, both before (baseline) and after ingestion of acetylsalicylic acid. The permeability response represents the difference between the two tests. A ratio above the 95th percentile for controls was classified as abnormal. RESULTS: Baseline permeability was higher in patients and spouses than in controls. An abnormal baseline permeability was seen in 36% of the patients, 23% of the spouses, 18% of the relatives, and 3% of the controls. After ingestion of acetylsalicylic acid, permeability increased significantly in all groups. Relatives were similar to patients with regard to permeability after exposure to acetylsalicylic acid, whereas spouses were similar to controls. The proportions with an abnormal permeability response to acetylsalicylic acid were 32% in patients, 14% in spouses, 41% in relatives, and 3% in controls. CONCLUSION The findings suggest that baseline permeability is determined by environmental factors, whereas permeability provoked by acetylsalicylic acid is a function of the genetically determined state of the mucosal barrier, and support the notion that environmental and hereditary factors interact in the pathogenesis of Crohn's disease. PMID- 9862834 TI - No relationship between gastric pH, small bowel bacterial colonisation, and diarrhoea in HIV-1 infected patients. AB - BACKGROUND/AIMS: Conclusive studies of small bowel bacterial overgrowth in patients with HIV-1 infection are limited. The relation was therefore determined between the quantity and species of bacteria in the proximal small intestine of HIV-1 infected patients and the presence of diarrhoea, gastric acidity, severity of immune deficiency, and clinical outcome. METHODS: Bacteria in the duodenal fluids obtained endoscopically from 32 HIV-1 infected patients, 21 of whom had diarrhoea, and seven control subjects without HIV-1 risk factors were quantified and speciated. Gastric pH was determined at the time of endoscopy. Clinical follow up was performed to assess outcome. RESULTS: Oropharyngeal Gram positive cocci were present in fluids from 28 patients (88%). Gram negative aerobic or facultatively anaerobic bacteria were present in fluids from 12 patients (38%), and strict anaerobes were detected in six patients (19%), but for both groups colony counts infrequently exceeded 10(4) colony forming units/ml. The number and species of bacteria did not correlate with the presence of diarrhoea, gastric pH, or CD4 lymphocyte count. CONCLUSIONS: Small bowel bacterial overgrowth is not common in HIV-1 infected patients, regardless of the presence of diarrhoea, and is not associated with hypochlorhydria PMID- 9862835 TI - Scintigraphic validation of a magnetic resonance imaging method to study gastric emptying of a solid meal in humans. AB - BACKGROUND: We have previously used a magnetic resonance imaging (MRI) method to study gastric emptying of liquids. So far, however, it has not possible to assess solid gastric emptying with this technique. AIMS: To validate scintigraphically MRI as a method for measuring emptying of a mixed solid/liquid meal. METHODS: In eight healthy subjects, gastric emptying of a solid/liquid (SM) and a liquid meal (LM) of identical energy content and macronutrient composition was studied by scintigraphy and MRI for 120 minutes. RESULTS: MRI and scintigraphy agreed with respect to emptying profiles (intraclass correlation coefficient (RI) SM: 0.988, RI, LM: 0.917), t1/2 (SMMRI: 129 (9), SMScinti: 123 (11) minutes, NS; LMMRI: 100 (7), LMScinti: 110 (8) minutes, NS) and AUC (SMMRI: 8999 (232), SMScinti: 8788 (277) min%, NS; LMMRI: 8819 (368), LMScinti: 8891 (321) min%, NS). CONCLUSIONS: MRI can be used to measure reliably gastric emptying not only of liquid but also of mixed solid/liquid meals in humans. PMID- 9862836 TI - Transabdominal bowel sonography for the detection of intestinal complications in Crohn's disease. AB - BACKGROUND: The course of Crohn's disease is characterised by the occurrence of intestinal complications such as strictures, intra-abdominal fistulas, or abscesses. Standard diagnostic procedures may fail to show these complications, in particular fistulas. AIMS: To test the value of transabdominal bowel sonography (T) for the detection of intestinal complications in Crohn's disease. METHODS: T was prospectively performed in 213 patients with Crohn's disease in a university based inflammatory bowel disease referral centre. Thirty three underwent resective bowel surgery and were included in this study. The accuracy of T to detect strictures, intra-abdominal fistulas, or abscesses was compared with surgical and pathological findings. RESULTS: T was able to identify strictures in 22/22 patients and to exclude it in 10/11 patients (100% sensitivity, 91% specificity). Fistulas were correctly identified in 20/23 patients and excluded in 9/10 patients (87% sensitivity, 90% specificity). Intra abdominal abscesses were correctly detected in 9/9 patients and excluded in 22/24 patients (100% sensitivity, 92% specificity). CONCLUSIONS: In experienced hands T is an accurate method for the detection of intestinal complications in Crohn's disease. T is thus recommended as a primary investigative method for evaluation of severe Crohn's disease. PMID- 9862837 TI - Use of magnetic resonance cholangiography in the diagnosis of choledocholithiasis: prospective comparison with a reference imaging method. AB - BACKGROUND: Magnetic resonance cholangiography (MRC) is a new technique for non invasive imaging of the biliary tract. AIM: To assess the results of MRC in patients with suspected bile duct stones as compared with those obtained with reference imaging methods. PATIENTS/METHODS: 70 patients (34 men and 36 women, mean (SD) age 71 (15.5) years; median 75) with suspected bile duct stones were included (cholangitis, 33; pancreatitis, three; suspected post-cholecystectomy choledocholithiasis, nine; cholestasis, six; stones suspected on ultrasound or computed tomography scan, 19). MR cholangiograms with two dimensional turbo spin echo sequences were acquired. Endoscopic retrograde cholangiography with or without sphincterotomy (n = 63), endosonography (n = 5), or intraoperative cholangiography (n = 2) were the reference imaging techniques used for the study and were performed within 12 hours of MRC. Radiologists were blinded to the results of endoscopic retrograde cholangiography and previous investigations. RESULTS: 49 patients (70%) had bile duct stones on reference imaging (common bile duct, 44, six of which impacted in the papilla; intrahepatic, four; cystic duct stump, one). Stone size ranged from 1 to 20 mm (mean 6.1, median 5.5). Twenty seven patients (55%) had bile duct stones smaller than 6 mm. MRC diagnostic accuracy for bile duct lithiasis was: sensitivity, 57.1%; specificity, 100%; positive predictive value, 100%; negative predictive value, 50%. CONCLUSIONS: Stones smaller than 6 mm are still often missed by MRC when standard equipment is used. The general introduction of new technical improvements is needed before this method can be considered reliable for the diagnosis of bile duct stones. PMID- 9862838 TI - Do gastroenterologists want to be trained in ultrasound? A national survey of trainees in gastroenterology. AB - AIMS: (1) To establish whether gastroenterologists wish to train in abdominal ultrasound according to the Royal College of Radiologists' document, Guidance for the training in ultrasound of medical non-radiologists. (2) To determine whether the ultrasound workload generated by gastroenterologists differs from that by other clinicians. METHODS: A postal questionnaire was sent to all 278 gastroenterology trainees. The indications and findings of 100 consecutive gastroenterologist requested scans were compared with 100 scans requested sequentially by other clinicians through a teaching hospital radiology department. RESULTS: 82% of the survey forms were returned. 77% of trainees wished to train in abdominal ultrasound and 68% were prepared to train in the manner outlined in the guideline document. However, 86% felt that they would ideally prefer not to assess renal or pelvic pathology, restricting to hepatobiliary diagnosis only. 73% of trainees did not anticipate that a further scan by a radiologist would be required. Comparison of gastroenterology scans with those requested by other clinicians revealed a relative excess of hepatobiliary indications and findings, and a notable paucity of renal and pelvic pathology in gastroenterology practice. CONCLUSIONS: There is general interest in abdominal ultrasound training among gastroenterology trainees and broad acceptance of the guideline document. However, most trainees perceive a focus of training restricted to hepatobiliary disease to be most appropriate. The case mix study provides support for this viewpoint. It is suggested that a more focused ultrasound training for gastroenterologists be considered. PMID- 9862839 TI - Inhibitory effect of oestradiol on activation of rat hepatic stellate cells in vivo and in vitro. AB - BACKGROUND: Hepatic stellate cells play a key role in the pathogenesis of hepatic fibrosis. AIMS: To examine the inhibitory effect of oestradiol on stellate cell activation. METHODS: In vivo, hepatic fibrosis was induced in rats by dimethylnitrosamine or pig serum. In vitro, rat stellate cells were activated by contact with plastic dishes resulting in their transformation into myofibroblast like cells. RESULTS: In the dimethylnitrosamine and pig serum models, treatment with oestradiol at gestation related doses resulted in a dose dependent suppression of hepatic fibrosis with restored content of hepatic retinyl palmitate, reduced collagen content, lower areas of stellate cells which express alpha smooth muscle actin (alpha-SMA) and desmin, and lower procollagen type I and III mRNA levels in the liver. In cultured stellate cells, oestradiol inhibited type I collagen production, alpha-SMA expression, and cell proliferation. These findings suggest that oestradiol is a potent inhibitor of stellate cell transformation. CONCLUSION: The antifibrogenic role of oestradiol in the liver may contribute to the sex associated differences in the progression from hepatic fibrosis to cirrhosis PMID- 9862841 TI - Death receptors in liver biology and pathobiology. PMID- 9862840 TI - Oral tolerance in disease. PMID- 9862842 TI - Regulation of electrogenic anion secretion in normal and cystic fibrosis gallbladder mucosa. AB - Fluid and ion transport across biliary epithelium contributes to bile flow. Alterations of this function may explain hepatobiliary complications in cystic fibrosis (CF). We investigated electrogenic anion transport across intact non-CF and CF human gallbladder mucosa in Ussing-type chambers. In non-CF tissues, baseline transmural potential difference (PD), short-circuit current (Isc), and resistance (R) were -2.2 +/- 0.3 mV (lumen negative), 40.7 +/- 7.8 microA/cm2, and 66.5 +/- 9.6 Omega. cm2, respectively (n = 14). The addition of forskolin (10(-5) mol/L) to the apical and basolateral baths and that of adenosine 5' triphosphate (ATP) (10(-4) mol/L) to the apical bath induced significant increases in Isc by 8.0 +/- 1.4 and 10.3 +/- 1.8 microA/cm2, respectively. Depletion of bathing solutions in Cl- and HCO3- significantly reduced baseline Isc and the forskolin- and ATP-induced increases in Isc. Anion secretion was stimulated by extracellular ATP via P2Y2 purinoceptors, as indicated by the effects of different nucleotides on Isc and on 36Cl efflux in cultured gallbladder epithelial cells. This effect was mediated by cytosolic calcium increase and Ca2+/calmodulin-dependent protein kinase II, as ascertained by using inhibitors. In CF preparations, basal PD and Isc were lower than in non-CF, and the response to forskolin was abolished, whereas the response to ATP was enhanced (P <.05 for all). We conclude that electrogenic anion secretion occurs in human gallbladder mucosa under basal state and is stimulated by an adenosine 3',5' cyclic monophosphate (cAMP)-dependent pathway mediated by cystic fibrosis transmembrane conductance regulator (CFTR), and by exogenous ATP via a CFTR independent pathway that is up-regulated in CF and involves P2Y2 purinoceptors and a calcium-dependent pathway. PMID- 9862843 TI - Effects of chronic ethanol administration on rat liver proteasome activities: relationship with oxidative stress. AB - We previously reported that ethanol elicits an increased protein oxidation in the liver of rats receiving chronic ethanol by continuous intragastric infusion (Tsukamoto-French method). This accumulation of oxidized proteins could result from a decrease in the cytosolic proteolysis, related specifically to alkaline protease and its major components, the proteasomes. Because several studies suggest that intracellular proteolysis depends on the severity of oxidative stress, we investigated the cytosolic proteolytic activity under two chronic ethanol treatment paradigms associated with varying degrees of oxidative stress. For 4 weeks, male rats received chronic ethanol by continuous intragastric infusion or by oral administration (10% ethanol ad libitum as sole drinking fluid). A significant decrease was evident for alkaline protease activity as well as for sodium dodecyl sulfate (SDS)-activated latent 20S proteasome (chymotrypsine-like [ChT-L] and peptidylglutamyl peptide hydrolase [PGPH] activities) in the liver of rats receiving ethanol by continuous intragastric infusion. Free radical production and related processes appeared to be contributing events in proteolysis inhibition, because phenethyl isothiocyanate (PIC), an inhibitor of cytochrome P4502E1 (CYP2E1), reduced the inhibition of the ethanol-related ChT-L activity. Moreover, the lipid peroxidation level was inversely correlated with ChT-L activity. In contrast, no such changes were observed in ChT-L and PGPH activities or in cellular free radical targets following the oral ad libitum consumption of 10% ethanol. It appears, thus, that only the alcohol treatment paradigm associated with an overt oxidative stress produced a significant inhibition of the proteasome activity. The mechanisms of proteasome inhibition could involve the formation of an endogenous inhibitor such as protein aggregates or aldehyde-derivative peptides. Whatever the mechanism, the inhibition of cytosolic proteolysis and the subsequent accumulation of damaged proteins may be involved in the oxidatively challenged alcoholic livers and play a pathogenic role in experimental alcoholic liver disease. PMID- 9862844 TI - Cost-effectiveness of ursodeoxycholic acid therapy in primary biliary cirrhosis. AB - Ursodeoxycholic acid (UDCA) is a safe and effective treatment for patients with primary biliary cirrhosis (PBC), but the cost of this drug has raised concerns regarding cost-effectiveness. The aim of our study was to determine the cost effectiveness of UDCA in PBC. We compared the costs and outcomes of managing PBC patients with and without UDCA. From two previously published trials, the effectiveness of UDCA was determined by comparing the annual reduction in the development of ascites, varices, variceal bleeding, encephalopathy, liver transplantation, and death between the treatment groups. Average annual costs for each of these events were estimated based on literature and institutional data. Approximately twice as many major events occurred in the placebo group compared with the UDCA group. The relative risk (RR) of liver transplantation (1.95; 95% CI: 1.14-3.68) and development of esophageal varices (3. 11; 95% CI: 1.57-10.65) were significantly higher in the placebo group compared with the UDCA group. There were no significant increases in the RR of ascites, variceal bleeding, encephalopathy, or death between the two groups. Based on the estimated annual cost of managing these events and the annual costs of UDCA ($2,500), there was an annual cost savings per patient of $1,372. Compared with the placebo group, patients receiving UDCA had a lower incidence of major complications and lower medical care costs. PMID- 9862845 TI - Transjugular intrahepatic portosystemic shunt versus endoscopic sclerotherapy for the prevention of variceal rebleeding after recent variceal hemorrhage. AB - Variceal hemorrhage continues to be a major cause of morbidity and mortality in cirrhotic patients. Transjugular intrahepatic portosystemic shunt (TIPS) is gaining wide acceptance as a treatment for several complications of portal hypertension. The aim of the current randomized study was to compare the transjugular shunt and endoscopic sclerotherapy (ES) for the prevention of variceal rebleeding (VB) in cirrhotic patients. Forty-six consecutive cirrhotic patients with variceal bleeding were randomly allocated to receive either transjugular shunt (22 patients) or ES (24 patients) 24 hours after control of bleeding. VB (50% vs. 9%) and early (first 6 weeks) VB (33% vs. 5%) were significantly more frequent in sclerotherapy patients; the actuarial probability of being free of VB was higher in the shunt group (P <.002). Eight patients (33%) of the sclerotherapy group and 3 patients (15%) of the shunt group died; the actuarial probability of survival was higher for the shunted patients (P <.05); 6 patients in the sclerotherapy group and none in the shunt group died from VB (P <.05). No difference was found in the proportion of patients with clinically evident hepatic encephalopathy (HE). These results show that the transjugular shunt is more effective than sclerotherapy in the prevention of both early and long-term VB. Moreover, a significant improvement in survival was found in the shunt group. PMID- 9862846 TI - Platelet aggregation and platelet-derived growth factor inhibition for prevention of insufficiency of the transjugular intrahepatic portosystemic shunt: a randomized study comparing trapidil plus ticlopidine with heparin treatment. AB - Intimal proliferation at the interface between prosthetic material and tissue is an intrinsic phenomenon of stenting and the major cause of insufficiency of the transjugular intrahepatic portosystemic shunt (TIPS). For its prevention, a randomized study was performed comparing standard heparin treatment with a combination of trapidil, a drug with anti-platelet-derived growth factor (PDGF) activity, and ticlopidine, a platelet aggregation inhibitor. Ninety patients with cirrhosis who received a transjugular shunt were randomized, and 84 patients completed the trial. Group 1 (n = 42) received a bolus of heparin (12 to 24 U/kg) at shunt placement, followed by 1 week of intravenous and 4 weeks of subcutaneous heparin treatment. Group 2 (n = 42) received the same heparin bolus, followed by a 1-day intravenous heparin treatment and a 6-month treatment with trapidil (400 mg/d) and ticlopidine (250 mg/d). Shunt function was assessed by duplex sonography and angiography. Stenoses were classified according to their location as type 1 (within the stent) and type 2 (in the draining hepatic vein). The estimated rate of overall stenoses (intention-to-treat analysis) at 1 year showed a significant reduction in patients receiving trapidil and ticlopidine (group 2) as compared with heparin (33 vs. 57%; P =.047). There was no difference in the estimated 1-year rate of type 1 stenoses between the two groups, but there was a significant reduction in type 2 stenoses (group 1: 58%, group 2: 19%; P =.016). The treatment effect continued after withdrawal of the drugs and was accompanied by a decreased incidence of rebleeding. The study demonstrates that the incidence of type 2 stenosis of the transjugular shunt can be reduced by combined inhibition of platelet aggregation and PDGF activity. The findings may be of relevance not only for the transjugular shunt, but also for other stent applications, e.g., vascular and biliary, as well as for bypass and shunt surgery. PMID- 9862847 TI - Clinical significance of serum bilirubin levels under ursodeoxycholic acid therapy in patients with primary biliary cirrhosis. AB - We determined whether the normalization of serum bilirubin level (SBL) induced by ursodeoxycholic acid (UDCA) therapy was associated with an improved clinical outcome in patients with primary biliary cirrhosis (PBC). We estimated the prognostic values of SBL measured after 6 months of UDCA treatment for survival free of orthotopic liver transplantation (OLT). We used a database of 548 patients with PBC followed in three trials of UDCA. Among UDCA-treated patients, we compared survival free of OLT in patients with normalized SBL (90%) and CC531s cells, a rat colon carcinoma (CC) cell line. Pit cells induced apoptosis in CC531s cells as shown by quantitative DNA fragmentation, agarose gel electrophoresis, and different modes of microscopy. When extracellular Ca2+ was chelated by ethylene glycol-bis(beta-aminoethyl ether)-N,N,-tetraacetic acid (EGTA) during coincubation or when the pit cells were preincubated with the granzyme inhibitor 3,4-dichloroisocoumarin (DCI), the induction of apoptosis was abolished. These results show that pit cells use the Ca2+-dependent perforin/granzyme pathway to induce apoptosis in the CC531s cells, and not the alternative Ca2+-independent Fas pathway. To further exclude the possibility of the involvement of the Fas pathway, we treated CC531s cells with recombinant Fas ligand. This treatment did not result in the induction of apoptosis, indicating that CC531s cells are resistant to Fas-mediated apoptosis. We conclude therefore that pit cells induce apoptosis in CC cells in vitro by the perforin/granzyme pathway. PMID- 9862850 TI - Propensity of ectopic liver to hepatocarcinogenesis: case reports and a review of the literature. AB - Two patients with ectopic liver are described. In one patient, a small ectopic liver attached to the gastric serosa developed hepatocellular carcinoma (HCC). The preoperative diagnosis was an alpha-fetoprotein (AFP)-producing carcinoma and a malignant ulcer of the stomach. Total gastrectomy and esophago-jejunostomy were performed. The tumor that measured 4 x 2 x 2 cm contained an AFP-producing HCC and normal liver tissue. In another patient who had alcoholic cirrhosis, ectopic liver on the serosa of the gallbladder was found to have the same histological changes as the mother liver. A survey of the literature disclosed more than 20 cases in which HCC developed outside the liver; the liver did not have HCC. By contrast, there was only one report on HCC occurring in the liver in the presence of a noncancerous, relatively large accessory liver lobe. Because ectopic liver does not have a complete vascular and ductal system as a normal liver, it is perhaps functionally handicapped and more prone to hepatocarcinogenesis. PMID- 9862852 TI - Role of beta1 integrins in adhesion and invasion of hepatocellular carcinoma cells. AB - To investigate the role of integrins in hepatocellular carcinoma (HCC) invasion, we analyzed the relationship between the expression and activity of beta1 integrins and the invasive ability of multiple HCC cell lines. Human HCC cell lines, PLC/PRF/5, Hep3B, HepG2, HLE, HuH7, and C3A cells, had high expression of beta1 and alpha6 subunits, and various levels of alpha1, alpha2, alpha3, and alpha5 expression as determined by cell surface flow cytometry. Activity of beta1 integrins was evaluated by cell adhesion to collagen, fibronectin, and laminin in the presence or absence of the stimulatory anti-beta1 monoclonal antibody (mAb) TS2/16. Different types of HCC cells showed various levels of constitutive activity of beta1 integrins as assessed by the TS2/16 requirement in cell adhesion. TS2/16 rapidly stimulated constitutively inactive or partially active beta1 integrins to fully active states, and as the result, the levels of cell adhesion to each ligand correlated with the expression levels of corresponding beta1 integrins. Thus, in the presence of TS2/16 stimulation, the levels of cell adhesion to collagen, fibronectin, and laminin correlated predominantly with the expression levels of alpha2, alpha5, and alpha6, respectively. Remarkably, as a result of in vitro chemoinvasion assay, the levels of constitutive activity of beta1 integrins correlated with the invasive ability of HCC cells. The inhibitory anti-beta1 mAb 13 almost completely blocked the invasion of PLC/PRF/5 and Hep3B cells that are the most invasive HCC cell lines. Alternatively, the stimulatory anti-beta1 mAb TS2/16 strongly inhibited the invasion. These results not only show an essential role of beta1 integrins in invasion of HCC cells but also suggest subtle regulatory mechanisms of cell invasion. PMID- 9862851 TI - Natural history of untreated nonsurgical hepatocellular carcinoma: rationale for the design and evaluation of therapeutic trials. AB - This study analyzed the natural history and prognostic factors of patients with nonsurgical hepatocellular carcinoma (HCC). Twenty variables from 102 cirrhotic patients with HCC who were not treated within prospective randomized controlled trials (RCT) were investigated through uni- and multivariate analyses. None of them was suitable for radical therapies (surgical resection, liver transplantation, or ethanol injection) or presented end-stage disease as reflected by an Okuda stage 3 or a Performance Status >/=3. Sixty-five patients were Child-Pugh A, 34 were B, and 3 were C. Most of them exhibited a preserved Performance Status Test (PST) (0 = 56; 1 = 38; 2 = 8). Tumor was solitary in 26 (600 ms) and fast inhibitory postsynaptic potentials (IPSPs; 5-50 ms) also were evoked. Application of gamma-aminobutyric acid-A (GABAA) antagonists blocked the fast inhibition and dramatically increased the firing rate response to StF tetanic stimuli. GABAA antagonists also increased the amplitude of the slow IPSP. The slow IPSP was reduced by GABAB antagonists. Blockade of excitatory amino acid (EAA) synaptic transmission allowed the monosynaptic bipolar-cell-mediated inhibition to be studied in isolation: EAA antagonists blocked most of the EPSP response to StF stimulation leaving fast and (an increased amplitude) slow IPSP components. The bipolar-cell IPSPs were mediated by GABAA and GABAB receptors as they were sensitive to GABAA and GABAB antagonists. The bipolar-cell IPSPs scaled with stimulation rate (20-400 Hz), reaching a maximum amplitude at 200 Hz. Inhibitory efficacy of bipolar-cell slow IPSPs were tested by their ability to reduce spiking in the face of sustained or brief current pulses. Established spike trains (by sustained injected current) were little affected by the onset of the slow IPSP. Weak brief currents injected during the slow IPSP were strongly inhibited. Strong brief currents could overcome the slow IPSP inhibitory effect. Inhibition was observed to interact with the intrinsic IA-like K+ currents to produce a complex control of cell spiking. Hyperpolarizing inhibition removes inactivation of IA to prevent subsequent inputs from driving the cell to threshold. Established depolarizing inputs, having allowed IA to inactivate, enable the cell to be highly sensitive to further depolarizing input. The term "conditional inhibition" is proposed to describe the general phenomenon where synaptic inhibition interacts with voltage sensitive intrinsic currents. PMID- 9862917 TI - Distal versus proximal inhibitory shaping of feedback excitation in the electrosensory lateral line lobe: implications for sensory filtering. AB - Distal versus proximal inhibitory shaping of feedback excitation in the electrosensory lateral line lobe: implications for sensory filtering. J. Neurophysiol. 80: 3214-3232, 1998. The inhibition controlling the indirect descending feedback (parallel fibers originating from cerebellar granule cells in the eminentia posterior pars granularis) to electrosensory lateral line lobe (ELL) pyramidal cells was studied using intracellular recording techniques in vitro. Parallel fibers (PF) contact stellate cells and dendrites of ventral molecular layer (VML) GABAergic interneurons. Stellate cells provide local input to pyramidal cell distal dendrites, whereas VML cells contact their somata and proximal dendrites. Single-pulse stimulation of PF evoked graded excitatory postsynaptic potentials (EPSPs) that were blocked by alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid and N-methyl--aspartate (NMDA) antagonists. The EPSPs peaked at 6.4 +/- 1.8 ms (mean +/- SE; n = 11) but took >50 ms to decay completely. Tetanic stimulation (100 ms, 100 Hz) produced a depolarizing wave with individual EPSPs superimposed. The absolute amplitude of the individual EPSPs decreased during the train. Spike rates, established by injected current, mostly were increased, but in some cells were decreased, by tetanic stimulation. Global application of a gamma-aminobutyric acid-A (GABAA) antagonist to the recorded cell's soma and apical dendritic region increased the EPSP peak and decay phase amplitudes. Tetanic stimulation always increased current-evoked spike rates after GABAA blockade during, and for several hundred milliseconds after, the stimulus. Application of a GABAB antagonist did not have any significant effects on the PF-evoked response. This, and the lack of any long hyperpolarizing inhibitory postsynaptic potentials, suggests that VML and stellate cell inhibition does not involve GABAB receptors. Focal GABAA antagonist applications to the dorsal molecular layer (DML) and pyramidal cell layer (PCL) had contrasting effects on PF-evoked EPSPs. DML GABAA blockade significantly increased the EPSP peak amplitude but not the decay phase of the EPSP, whereas PCL GABAA-blockade significantly increased the decay phase, but not the EPSP peak, amplitude. The order of antagonist application did not affect the outcome. On the basis of the known circuitry of the ELL, we conclude that the distal inhibition originated from GABAergic molecular layer stellate cells and the proximal inhibition originated from GABAergic cells of the ventral molecular layer (VML cells). Computer modeling of distal and proximal inhibition suggests that intrinsic differences in IPSP dynamics between the distal and proximal sites may be amplified by voltage-dependent NMDA receptor and persistent sodium currents. We propose that the different time courses of stellate cell and VML cell inhibition allows them to act as low- and high-pass filters respectively on indirect descending feedback to ELL pyramidal cells. PMID- 9862918 TI - Different VAMP/synaptobrevin complexes for spontaneous and evoked transmitter release at the crayfish neuromuscular junction. AB - Different VAMP/synaptobrevin complexes for spontaneous and evoked transmitter release at the crayfish neuromuscular junction. J. Neurophysiol. 80: 3233-3246, 1998. Although vesicle-associated membrane protein (VAMP/synaptobrevin) is essential for evoked neurotransmitter release, its role in spontaneous transmitter release remains uncertain. For instance, many studies show that tetanus toxin (TeNT), which cleaves VAMP, blocks evoked transmitter release but leaves some spontaneous transmitter release. We used recombinant tetanus and botulinum neurotoxin catalytic light chains (TeNT-LC, BoNT/B-LC, and BoNT/D-LC) to examine the role of VAMP in spontaneous transmitter release at neuromuscular junctions (nmj) of crayfish. Injection of TeNT-LC into presynaptic axons removed most of the VAMP immunoreactivity and blocked evoked transmitter release without affecting nerve action potentials or Ca2+ influx. The frequency of spontaneous transmitter release was little affected by the TeNT-LC when the evoked transmitter release had been blocked by >95%. The spontaneous transmitter release left after TeNT-LC treatment was insensitive to increases in intracellular Ca2+. BoNT/B-LC, which cleaves VAMP at the same site as TeNT-LC but uses a different binding site, also blocked evoked release but had minimal effect on spontaneous release. However, BoNT/D-LC, which cleaves VAMP at a different site from the other two toxins but binds to the same position on VAMP as TeNT, blocked both evoked and spontaneous transmitter release at similar rates. The data indicate that different VAMP complexes are employed for evoked and spontaneous transmitter release; the VAMP used in spontaneous release is not readily cleaved by TeNT or BoNT/B. Because the exocytosis that occurs after the action of TeNT cannot be increased by increased intracellular Ca2+, the final steps in neurotransmitter release are Ca2+ independent. PMID- 9862919 TI - Both supplementary and presupplementary motor areas are crucial for the temporal organization of multiple movements. AB - Both supplementary and presupplementary motor areas are crucial for the temporal organization of multiple movements. J. Neurophysiol. 80: 3247-3260, 1998. To study the involvement of the supplementary (SMA) and presupplementary (pre-SMA) motor areas in performing sequential multiple movements that are individually separated in time, we injected muscimol, a gamma-aminobutyric acid agonist, bilaterally into the part of each area that represents the forelimb. Two monkeys were trained to perform three different movements, separated by a waiting time, in four or six different orders. First, each series of movements was learned during five trials guided by visual signals that indicated the correct movements. The monkeys subsequently executed the three movements in the memorized order, without the visual signals. After the injection of muscimol (3 microliter, 5 micrograms/microliters in 10 min) into either the SMA or pre-SMA bilaterally, the animals started making errors in performing the sequence of movements correctly from memory. However, when guided with a visual signal, they could select and perform the three movements correctly. The impaired memory-based sequencing of movements worsened progressively with time until the animals could not perform the task. Yet they still could associate the visual signals with the different movements at that stage. In control experiments on two separate monkeys, we found that injections of the same amount of muscimol into either the SMA or pre-SMA did not cause problems with nonsequential reaching movement regardless of whether it was visually triggered or self-initiated. These results support the view that both the SMA and pre-SMA are crucially involved in sequencing multiple movements over time. PMID- 9862920 TI - Contribution of supragranular layers to sensory processing and plasticity in adult rat barrel cortex. AB - Contribution of supragranular layers to sensory processing and plasticity in adult rat barrel cortex. J. Neurophysiol. 80: 3261-3271, 1998. In mature rat primary somatic sensory cortical area (SI) barrel field cortex, the thalamic recipient granular layer IV neurons project especially densely to layers I, II, III, and IV. A prior study showed that cells in the supragranular layers are the fastest to change their response properties to novel changes in sensory inputs. Here we examine the effect of removing supragranular circuitry on the responsiveness and synaptic plasticity of cells in the remaining layers. To remove the layer II + III (supragranular) neurons from the circuitry of barrel field cortex, N-methyl--aspartate (NMDA) was applied to the exposed dura over the barrel cortex, which destroys those neurons by excitotoxicity without detectable damage to blood vessels or axons of passage. Fifteen days after NMDA treatment, the first responsive cells encountered were 400-430 micrometers below the pial surface. In separate cases triphenyltetrazolium chloride (TTC), a vital dye taken up by living cells, was absent from the lesion area. Cytochrome oxidase (CO) activity was absent in the first few tangential sections through the barrel field in all cases before arriving at the CO-dense barrel domains. These findings indicate that the lesions were quite consistent from animal to animal. Controls consisted of applying vehicle without NMDA under similar conditions. Responses of D2 barrel cells were assessed for spontaneous activity and level of response to stimulation of the principal D2 whisker and four surround whiskers D1, D3, C2, and E2. In two additional groups of animals treated in the same way, sensory plasticity was assessed by trimming all whiskers except D2 and either D1 or D3 (called Dpaired) for 7 days before recording cortical responses. Such whisker pairing normally potentiates D2 barrel cell responses to stimulation of the two intact whiskers (D2 + Dpaired). After NMDA lesions, cortical cells still responded to all whiskers tested. Cells in lesioned cortex showed reduced response amplitude compared with sham-operated controls to all D-row whiskers. In arc surround whisker (C2 or E2) responses were normal. Spontaneous activity did not change significantly in any remaining layer at the time tested. Modal latencies to stimulation of principal D2 or surround D1 or D3 whiskers showed no significant change after lesioning. These findings indicate that there is a reasonable preservation of the response properties of layer IV, V, VI neurons after removal of layer II-III neurons in this way. Whisker pairing plasticity in layer IV-VI D2 barrel column neurons occurred in both lesioned and sham animals but was reduced significantly in lesioned animals compared with controls. The response bias generated by whisker trimming (Dpaired/Dcut + Dpaired ratio) was less pronounced in NMDA-lesioned than sham-lesioned animals. Proportionately fewer neurons in layer IV (52 vs. 64%) and in the infragranular layers (55 vs. 68%) exhibited a clear response bias to paired whiskers. We conclude that receptive-field plasticity can occur in layers IV-VI of barrel cortex in the absence of the supragranular layer circuitry. However, layer I-III circuitry does play a role in normal receptive-field generation and is required for the full expression of whisker pairing plasticity in granular and infragranular layer cells. PMID- 9862922 TI - Locomotor modulation of disynaptic EPSPs from the mesencephalic locomotor region in cat motoneurons. AB - Locomotor modulation of disynaptic EPSPs from the mesencephalic locomotor region in cat motoneurons. J. Neurophysiol. 80: 3284-3296, 1998. When low-frequency tetanization of the mesencephalic locomotor region (MLR) produce fictive locomotion in unanesthetized, decerebrate cats, each MLR stimulus produces a distinctive cord dorsum potential (CDP) and oligosynaptic excitatory postsynaptic potentials (EPSPs) in many lumbosacral motoneurons. The average segmental latency from the initial CDP wave [mean delay from stimulus: 4.3 +/- 0.9 (SD) ms] to the onset of detectable MLR EPSPs was 1.6 +/- 0.4 ms, suggesting a disynaptic segmental connection. In gastrocnemius/soleus, flexor hallucis longus, flexor digitorum longus, tibialis anterior, and posterior biceps-semitendinosus motoneurons (35/38 cells), MLR EPSPs either appeared or were enhanced during the phase of fictive stepping in which the target motoneurons were depolarized and the motor pool was active (the phase), with parallel changes between EPSP amplitudes and membrane depolarization. In contrast, MLR stimulation produced small (1/10) or no EPSPs in extensor digitorum longus (EDL) motoneurons, with no phase enhancement (4/10) or oligosynaptic inhibitory postsynaptic potentials during the phase (5/10). Eight of 10 flexor digitorum longus (FDL) cells exhibited membrane depolarization in the early flexion phase of fictive stepping, and five of these showed parallel enhancement of disynaptic MLR EPSPs during early flexion. Three cases were studied when the FDL motor pool exhibited exclusively extensor phase firing. In these cases, the disynaptic MLR EPSPs were enhanced only during the extensor phase, accompanied by membrane depolarizations. We conclude that the last-order interneurons that produce disynaptic MLR EPSPs may well participate in producing the depolarizing locomotor drive potentials (LDPs) found in hindlimb motoneurons during fictive locomotion. However, the absence of linkage between MLR EPSP enhancement and LDP depolarizations in EDL motoneurons suggests that other types of excitatory interneurons also must be involved at least in some motor pools. We compared these patterns with the modulation of disynaptic EPSPs produced in FDL cells by stimulation of the medial longitudinal fasciculus (MLF). In all seven FDL motoneurons tested, disynaptic MLF EPSPs appeared only during the extension phase, regardless of when the FDL motoneurons were active. The fact that the modulation patterns of MLR and MLF disynaptic EPSPs is different in FDL motoneurons indicates that the two pathways do not converge on common last-order interneurons to that motor pool. PMID- 9862921 TI - Response of anterior parietal cortex to different modes of same-site skin stimulation. AB - Response of anterior parietal cortex to different modes of same-site skin stimulation. J. Neurophysiol. 80: 3272-3283, 1998. Intrinsic optical signal (IOS) imaging was used to study responses of the anterior parietal cortical hindlimb region (1 subject) and forelimb region (3 subjects) to repetitive skin stimulation. Subjects were four squirrel monkeys anesthetized with a halothane/nitrous oxide/oxygen gas mixtures. Cutaneous flutter of 25 Hz evoked a reflectance decrease in the sectors of cytoarchitectonic areas 3b and/or 1 that receive input from the stimulated skin site. The intrinsic signal evoked by 25-Hz flutter attained maximal intensity 0.05, analysis of variance). Thus the results in present study indicate that both the spontaneous discharge and the driven response to rotation of pigeon HSCPAs recovered their normal physiological status between 30 and 150 days PIS after hair cell death due to aminoglycoside ototoxicity. The recovery was systematic for the parameters chosen to be tested with the exception of the cupula long time constant, tauL. The mechanisms (changes in ciliary dynamics, changes in hair cell ionic currents, changes in bouton terminals, etc.) underlying these changes await further morphophysiological studies. PMID- 9862924 TI - Temporal and intensity coding of pain in human cortex. AB - Temporal and intensity coding of pain in human cortex. J. Neurophysiol. 80:3312 3320, 1998. We used a high-resolution functional magnetic resonance imaging (fMRI) technique in healthy right-handed volunteers to demonstrate cortical areas displaying changes of activity significantly related to the time profile of the perceived intensity of experimental somatic pain over the course of several minutes. Twenty-four subjects (ascorbic acid group) received a subcutaneous injection of a dilute ascorbic acid solution into the dorsum of one foot, inducing prolonged burning pain (peak pain intensity on a 0-100 scale: 48 +/- 3, mean +/- SE; duration: 11.9 +/- 0.8 min). fMRI data sets were continuously acquired for approximately 20 min, beginning 5 min before and lasting 15 min after the onset of stimulation, from two sagittal planes on the medial hemispheric wall contralateral to the stimulated site, including the cingulate cortex and the putative foot representation area of the primary somatosensory cortex (SI). Neural clusters whose fMRI signal time courses were positively or negatively correlated (P < 0.0005) with the individual pain intensity curve were identified by cross-correlation statistics in all 24 volunteers. The spatial extent of the identified clusters was linearly related (P < 0.0001) to peak pain intensity. Regional analyses showed that positively correlated clusters were present in the majority of subjects in SI, cingulate, motor, and premotor cortex. Negative correlations were found predominantly in medial parietal, perigenual cingulate, and medial prefrontal regions. To test whether these neural changes were due to aspecific arousal or emotional reactions, related either to anticipation or presence of pain, fMRI experiments were performed with the same protocol in two additional groups of volunteers, subjected either to subcutaneous saline injection (saline: n = 16), inducing mild short-lasting pain (peak pain intensity 23 +/- 4; duration 2.8 +/- 0.6 min) or to nonnoxious mechanical stimulation of the skin (controls: n = 16) at the same body site. Subjects did not know in advance which stimulus would occur. The spatial extent of neural clusters whose signal time courses were positively or negatively correlated with the mean pain intensity curve of subjects injected with ascorbic acid was significantly larger (P < 0.001) in the ascorbic acid group than both saline and controls, suggesting that the observed responses were specifically related to pain intensity and duration. These findings reveal distributed cortical systems, including parietal areas as well as cingulate and frontal regions, involved in dynamic encoding of pain intensity over time, a process of great biological and clinical relevance. PMID- 9862925 TI - Functional reorganization of the rat motor cortex following motor skill learning. AB - Functional reorganization of the rat motor cortex following motor skill learning. J. Neurophysiol. 80: 3321-3325, 1998. Adult rats were allocated to either a skilled or unskilled reaching condition (SRC and URC, respectively). SRC animals were trained for 10 days on a skilled reaching task while URC animals were trained on a simple bar pressing task. After training, microelectrode stimulation was used to derive high resolution maps of the forelimb and hindlimb representations within the motor cortex. In comparison with URC animals, SRC animals exhibited a significant increase in mean area of the wrist and digit representations but a decrease in elbow/shoulder representation within the caudal forelimb area. No between-group differences in areal representation were found in either the hindlimb or rostral forelimb areas. These results demonstrate that motor skill learning is associated with a reorganization of movement representations within the rodent motor cortex. PMID- 9862926 TI - Stimulation of human thalamus for pain relief: possible modulatory circuits revealed by positron emission tomography. AB - Stimulation of human thalamus for pain relief: possible modulatory circuits revealed by positron emission tomography. J. Neurophysiol. 80: 3326-3330, 1998. Stimulation of the somatosensory thalamus was used for more than 2 decades to treat chronic pain in the human. However, despite clinical reports of successful results, little is known about the actual mechanisms mediating this form of stimulation-produced analgesia. To reveal possible neuronal pathways evoked by thalamic stimulation, we measured regional changes in cerebral blood flow (rCBF) in five patients who received successful long-term relief of chronic pain with somatosensory thalamic stimulation. Positron emission tomography during thalamic stimulation revealed significant activation of the thalamus in the region of the stimulating electrodes as well as activation of the insular cortex ipsilateral to the thalamic electrodes (contralateral to the patients' clinical pain). For these patients, thalamic stimulation also evoked paresthesiae that included thermal sensations in addition to tingling sensations. Results of this study indicate that in some cases somatosensory thalamic stimulation may activate a thalamocortical pain modulation circuit that involves thermal pathways. These results are consistent with other recent reports suggesting that activation of thermal pathways may contribute to modulation of nociceptive information. PMID- 9862927 TI - Frontal eye field neurons orthodromically activated from the superior colliculus. AB - Frontal eye field neurons orthodromically activated from the superior colliculus. J. Neurophysiol. 80: 3331-3333, 1998. Anatomical studies have shown that the frontal eye field (FEF) and superior colliculus (SC) of monkeys are reciprocally connected, and a physiological study described the signals sent from the FEF to the SC. Nothing is known, however, about the signals sent from the SC to the FEF. We physiologically identified and characterized FEF neurons that are likely to receive input from the SC. Fifty-two FEF neurons were found that were orthodromically activated by electrical stimulation of the intermediate or deeper layers of the SC. All the neurons that we tested (n = 34) discharged in response to visual stimulation. One-half also discharged when saccadic eye movements were made. This provides the first direct evidence that the ascending pathway from SC to FEF might carry visual- and saccade-related signals. Our findings support a hypothesis that the SC and the FEF interact bidirectionally during the events leading up to saccade generation. PMID- 9862929 TI - Influence of predictive information on responses of tonically active neurons in the monkey striatum. AB - Influence of predictive information on responses of tonically active neurons in the monkey striatum. J. Neurophysiol. 80: 3341-3344, 1998. We investigated how the expectation of a signal of behavioral significance influences the activity of tonically active neurons in the striatum of two monkeys performing a simple reaction time task under two conditions, an uncued condition in which the trigger stimulus occurred randomly in time and a cued condition in which the same trigger was preceded by an instruction stimulus serving as a predictive signal for the forthcoming signal eliciting an immediate behavioral reaction. Both monkeys benefited from the presence of the instruction stimulus to reduce their reaction time, suggesting an increased ability to predict the trigger onset during cued trials compared with uncued trials. A majority of neurons (199/272, 73%) showed a phasic reduction in activity after the onset of the trigger stimulus in the uncued condition, whereas only 38% responded to the same stimulus when it was preceded by the instruction. Furthermore, magnitudes of trigger responses in the uncued condition were significantly higher than in the cued condition. Fifty seven percent of the neurons responded to the instruction stimulus, and one-half of the neurons losing their response to the trigger in the cued condition responded to the instruction stimulus. These findings suggest that responses of tonic striatal neurons to a trigger stimulus for movement were influenced by predictive information. PMID- 9862928 TI - Glycine uptake governs glycine site occupancy at NMDA receptors of excitatory synapses. AB - Glycine uptake governs glycine site occupancy at NMDA receptors of excitatory synapses. J. Neurophysiol. 80: 3336-3340, 1998. At central synapses occupation of glycine binding sites of N-methyl--aspartate receptors (NMDA-Rs) is a necessary prerequisite for the excitatory neurotransmitter glutamate to activate these receptors. There is conflicting evidence as to whether glycine binding sites normally are saturated. If they are not, then alterations in local glycine concentration could modulate excitatory synaptic transmission. By using an in vitro brain stem slice preparation we investigated whether the glycine site is saturated for synaptically activated NMDA-Rs in neonatal rat hypoglossal motoneurons. We found that the NMDA-R-mediated component of spontaneous miniature excitatory postsynaptic currents could be potentiated by exogenously applied glycine as well as by -serine. The effects of glycine were observed only at concentrations (100 microM or more) two orders of magnitude above the apparent dissociation constant of glycine from NMDA receptors. In contrast, -serine, a nontransported NMDA-R glycine site agonist, was effective in the low micromolar range, i.e., at concentrations similar to those found to be effective on isolated cells or on outside-out patches. We conclude that at these synapses the glycine concentration around synaptic NMDA-Rs is set below the concentration required to saturate their glycine site and is likely to be stabilized by a powerful glycine transport mechanism. PMID- 9862930 TI - Slow covariations in neuronal resting potentials can lead to artefactually fast cross-correlations in their spike trains. AB - Slow covariations in neuronal resting potentials can lead to artefactually fast cross-correlations in their spike trains. J. Neurophysiol. 80: 3345-3351, 1998. A model of two lateral geniculate nucleus (LGN) cells, which interact only through slow (tens of seconds) covariations in their resting membrane potentials, is used here to investigate the effect of such covariations on cross-correlograms taken during stimulus-driven conditions. Despite the slow timescale of the interactions, the model generates cross-correlograms with peak widths in the range of 25-200 ms. These bear a striking resemblance to those reported in studies of LGN cells by Sillito et al., which were taken at the time as evidence of a fast spike timing synchronization interaction; the model highlights the possibility that those correlogram peaks may have been caused by a mechanism other than spike synchronization. Slow resting potential covariations are suggested instead as the dominant generating mechanism. How can a slow interaction generate covariogram peaks with a width 100-1,000 times thinner than its timescale? Broad peaks caused by slow interactions are modulated by the cells' poststimulus time histograms (PSTHs). When the PSTHs have thin peaks (e.g., tens of milliseconds), the cross-correlogram peaks generated by slow interactions will also be thin; such peaks are easily misinterpretable as being caused by fast interactions. Although this point is explored here in the context of LGN recordings, it is a general point and applies elsewhere. When cross correlogram peak widths are of the same order of magnitude as PSTH peak widths, experiments designed to reveal short-timescale interactions must be interpreted with the issue of possible contributions from slower interactions in mind. PMID- 9862931 TI - Modulation of calcium-dependent postsynaptic depression contributes to an adaptive sensory filter. AB - Modulation of calcium-dependent postsynaptic depression contributes to an adaptive sensory filter. J. Neurophysiol. 80: 3352-3355, 1998. The ability of organisms to ignore unimportant patterns of sensory input may be as critical as the ability to attend to those that are behaviorally relevant. Mechanisms used to reject irrelevant inputs range from peripheral filters, which allow only restricted portions of the spectrum of possible inputs to pass, to higher-level processes, which actively select stimuli to be "attended to." Recent studies of several lower vertebrates demonstrate the presence of adaptive sensory filters, which "learn," with a time course of a few minutes, to cancel predictable patterns of sensory input without compromising responses to novel stimuli. Predictable stimuli include "reafferent" stimuli, which occur as a result of an animal's own activity, as well as stimuli that are simply repetitive. The adaptive characteristic of these filters depends on an anti-Hebbian form of synaptic plasticity that modulates the strength of multisensory dendritic inputs resulting in the genesis of "negative image" signals, which cancel the predicted pattern of sensory afference. This report provides evidence that the mechanism underlying the anti-Hebbian plasticity involves the modulation of a calcium dependent form of postsynaptic depression. PMID- 9862932 TI - ATP P2x receptors and sensory synaptic transmission between primary afferent fibers and spinal dorsal horn neurons in rats. AB - ATP P2x receptors and sensory synaptic transmission between primary afferent fibers and spinal dorsal horn neurons in rats. J. Neurophysiol. 80: 3356-3360, 1998. Glutamate is a major fast transmitter between primary afferent fibers and dorsal horn neurons in the spinal cord. Recent evidence indicates that ATP acts as another fast transmitter at the rat cervical spinal cord and is proposed to serve as a transmitter for nociception and pain. Sensory synaptic transmission between dorsal root afferent fibers and neurons in the superficial dorsal horn of the lumbar spinal cord were examined by whole cell patch-clamp recording techniques. Experiments were designed to test if ATP could serve as a transmitter at the lumbar spinal cord. Monosynaptic excitatory postsynaptic currents (EPSCs) were completely abolished after the blockade of both glutamatergic alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid/kainate and N-methyl--aspartate receptors. No residual current was detected, indicating that glutamate but not ATP is a fast transmitter at the dorsal horn of the lumbar spinal cord. Pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), a selective P2x receptor antagonist, produced an inhibitory modulatory effect on fast EPSCs and altered responses to paired-pulse stimulation, suggesting the involvement of a presynaptic mechanism. Intrathecal administration of PPADS did not produce any antinociceptive effect in two different types of behavioral nociceptive tests. The present results suggest that ATP P2x2 receptors modulate excitatory synaptic transmission in the superficial dorsal horn of the lumbar spinal cord by a presynaptic mechanism, and such a mechanism does not play an important role in behavioral responses to noxious heating. The involvement of other P2x subtype receptors, which is are less sensitive to PPADS, in acute nociceptive modulation and persistent pain remains to be investigated. PMID- 9862933 TI - Spontaneous apamin-sensitive hyperpolarizations in dopaminergic neurons of neonatal rats. AB - Spontaneous apamin-sensitive hyperpolarizations in dopaminergic neurons of neonatal rats. J. Neurophysiol. 80: 3361-3364, 1998. Intracellular recordings from substantia nigra slices revealed the existence of spontaneous hyperpolarizations (amplitude 2-8 mV, duration 100-400 ms) at -60 mV in most dopaminergic neurons of neonatal (9-15 days) but not adult rats. These events were blocked by apamin (300 nM) and bicuculline methochloride (100-300 microM), which blocks apamin-sensitive currents. They were unaffected by the selective gamma-aminobutyric acid-A (GABAA) antagonists SR95531 (100 microM) and picrotoxin (30-50 microM), the GABAB antagonist CGP35348 (300 microM), the D2 antagonist haloperidol (1 microM), and the metabotropic antagonist MCPG (1 mM). The hyperpolarizations were strongly attenuated or abolished when recording electrodes contained 200 mM 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. They were resistant to tetrodotoxin in the majority of the cells. They had some voltage dependency and were in some cases transiently potentiated when cells were briefly depolarized by current injection. We conclude that dopaminergic neurons have developmentally regulated physiological properties. These spontaneous hyperpolarizations might affect the firing rate of these cells, which was found to be lower in neonates than in adults. PMID- 9862934 TI - Neuronal discriminator formed by metabotropic gamma-aminobutyric acid receptors. AB - Neuronal discriminator formed by metabotropic gamma-aminobutyric acid receptors. J. Neurophysiol. 80: 3365-3368, 1998. Neurotransmitters function in one of two modes, promoting either inhibition or excitation. However, the metabotropic gamma aminobutyric acid receptor (GABABR) system can switch between these modes. In the presence of a small excitatory stimulus, the GABABR mediates a shunting inhibition that suppresses excitation. However, in the presence of a strong excitatory stimulus, the GABABR potentiates the response. This bipartite action is accomplished by linking the GABABR to two electrogenic mechanisms; one activates an outward current and another reduces an outward current. As a consequence, the GABABR serves as a discriminator that reduces the influence of weak signals while augmenting responses to strong signals. In retinal ganglion cells, this mechanism acts to promote the communication of phasic information. PMID- 9862935 TI - Damping in reflexively active and areflexive lengthening muscle evaluated with inertial loads. AB - Damping in reflexively active and areflexive lengthening muscle evaluated with inertial loads. J. Neurophysiol. 80: 3369-3372, 1998. Studies of active areflexive muscle have shown that during a constant velocity stretch the increment in force elicited by an incremental length change falls dramatically after a few hundred micrometers of stretch, a finding labeled as "muscle yield." The mechanical behavior after the yield was like a viscous damper, in that force varied only with velocity. In light of these observations, our aims were to determine whether viscous properties are also evident under more physiological conditions, specifically under inertial loading, and to evaluate the damping action of reflexively intact compared with that of deafferented muscle. The active soleus muscle in a decerebrate cat was forcibly stretched by a simulated inertia with a specified initial velocity. We compared muscle length changes when afferent pathways were intact with those recorded after cutting the dorsal roots. Our findings were that areflexive muscle showed highly damped responses, with large changes in mean muscle length, indicative of high viscosity relative to stiffness. In contrast, reflexively active muscle produced lightly damped oscillations, with minimal changes in mean length, reflecting low viscosity and high stiffness. It appears that the stretch reflect modifies the relative contributions of elastic and viscous-like forces, maintaining elasticity, which in turn sustains oscillations. These differences highlight tradeoffs between positional and velocity regulation, in that elastic properties of reflexively active muscle promote oscillations with modest change in mean muscle length, whereas viscous-like properties of areflexive muscle produce damped responses, with poor positional regulation. PMID- 9862936 TI - New mechanism that accounts for position sensitivity of saccades evoked in response to stimulation of superior colliculus. AB - New mechanism that accounts for position sensitivity of saccades evoked in response to stimulation of superior colliculus. J. Neurophysiol. 80: 3373-3379, 1998. Electrical stimulation of the feline superior colliculus (SC) is known to evoke saccades whose size depends on the site stimulated (the "characteristic vector" of evoked saccades) and the initial position of the eyes. Similar stimuli were recently shown to produce slow drifts that are presumably caused by relatively direct projections of the SC onto extraocular motoneurons. Both slow and fast evoked eye movements are similarly affected by the initial position of the eyes, despite their dissimilar metrics, kinematics, and anatomic substrates. We tested the hypothesis that the position sensitivity of evoked saccades is due to the superposition of largely position-invariant saccades and position dependent slow drifts. We show that such a mechanism can account for the fact that the position sensitivity of evoked saccades increases together with the size of their characteristic vector. Consistent with it, the position sensitivity of saccades drops considerably when the contribution of slow drifts is minimal as, for example, when there is no overlap between evoked saccades and short-duration trains of high-frequency stimuli. PMID- 9862937 TI - NMDA-Induced intrinsic voltage oscillations depend on L-type calcium channels in spinal motoneurons of adult turtles. AB - NMDA-induced intrinsic voltage oscillations depend on L-type calcium channels in spinal motoneurons of adult turtles. J. Neurophysiol. 80: 3380-3382, 1998. In a slice preparation from adult turtles, bath-applied N-methyl-D-aspartate (NMDA) induced rhythmic activity in spinal motoneurons. The underlying intrinsic oscillation in membrane potential was revealed in the presence of tetrodotoxin (TTX). NMDA-induced rhythmicity, in the presence or absence of TTX, was abolished or reduced by NMDA receptor antagonists and by three different classes of antagonists for L-type calcium channels. It is suggested that both NMDA receptor channels and L-type calcium channels contribute to NMDA-induced intrinsic oscillations in mature spinal motoneurons. PMID- 9862938 TI - Brain-derived neurotrophic factor (BDNF) modulates inhibitory, but not excitatory, transmission in the CA1 region of the hippocampus. AB - Brain-derived neurotrophic factor (BDNF) modulates inhibitory, but not excitatory, transmission in the CA1 region of the hippocampus. J. Neurophysiol. 80: 3383-3386, 1998. Brain-derived neurotrophic factor (BDNF) has been reported to have rapid effects on synaptic transmission in the hippocampus. We report here that bath application of BDNF causes a small but significant decrease in stimulus evoked inhibitory postsynaptic currents (IPSCs) on CA1 pyramidal cells, which is prevented by the tyrosine kinase inhibitor lavendustin A. BDNF causes a decrease in the 1/CV2 of the IPSC, and also reduces paired-pulse depression of the IPSC, suggesting a presynaptic site of action. In contrast, BDNF did not have a detectable effect on field excitatory postsynaptic potentials measured in stratum radiatum. We conclude that BDNF has a selective depressant action on inhibitory transmission in the hippocampus, due at least in part to a presynaptic mechanism. PMID- 9862939 TI - Na+-gated nonselective cation channel from lobster olfactory projection neurons. AB - Na+-gated nonselective cation channel from lobster olfactory projection neurons. J. Neurophysiol. 80: 3387-3391, 1998. A nonselective cation channel specifically activated by intracellular Na+ was identified in cell-free patches taken from cultured lobster olfactory projection neurons. Na+ reversibly activates the channel in a concentration-dependent manner, with a "half-effect" Na+ concentration of 76.4 mM at -60 mV. The conductance of the channel is 32 pS. The channel is permeable to both alkali metal (Li+ > Na+ > K+ > Rb+ > Cs+) and divalent (Ca2+ > Mn2+ > Sr2+ > Mg2+ > Ba2+ > Na+) cations. The presence of a channel with the ability to generate plateau potentials suggests that the channel may potentially contribute to oscillatory behavior in these olfactory interneurons. PMID- 9862940 TI - Task-dependent selectivity of movement-related neuronal activity in the primate prefrontal cortex. AB - Task-dependent selectivity of movement-related neuronal activity in the primate prefrontal cortex. J. Neurophysiol. 80: 3392-3397, 1998. We studied movement related neuronal activity in the dorsolateral prefrontal cortex from the perspective of a general role for the prefrontal cortex in controlling motor behavior to achieve a specific goal according to the requirements of a given task. Monkeys were trained to perform two delayed motor tasks. The first task involved reaching for a target that matched the shape of a cue. The second task involved reaching for a target that matched the location of the cue. A majority (54%) of 175 movement-related prefrontal neurons exhibited preference for either the target shape or the type of task requirements. Sixty-four neurons (36%) were selectively active while reaching for a circle or a triangle. On the other hand, the activity of 59 neurons (34%) depended on whether the task required matching the shape or the location. These properties, characterizing the movement-related neuronal activity in the prefrontal cortex, rarely were found in the arm area of the primary motor cortex. Only 1 of 130 movement-related neurons (0.8%) showed task selectivity, and none showed target-shape selectivity. PMID- 9862941 TI - Symptomatic benefit from eradicating Helicobacter pylori infection in patients with nonulcer dyspepsia. AB - BACKGROUND: The eradication of Helicobacter pylori infection is beneficial in patients with gastric or duodenal ulcers. The value of eradicating the infection in patients with dyspepsia and no evidence of ulcer disease is not known. METHODS: We performed a randomized, placebo-controlled trial comparing the efficacy of treatment for two weeks with 20 mg of omeprazole orally twice daily, 500 mg of amoxicillin three times daily (with 500 mg of tetracycline three times daily substituted for amoxicillin in patients allergic to penicillin), and 400 mg of metronidazole three times daily (160 patients) with that of omeprazole alone (158 patients) for resolving symptoms of dyspepsia in patients with H. pylori infection but no evidence of ulcer disease on upper gastrointestinal endoscopy. Symptoms were assessed with the Glasgow Dyspepsia Severity Score, with resolution of symptoms defined as a score of 0 or 1 in the preceding six months (maximal score, 20). One year later the patients were assessed to determine the frequency of the resolution of symptoms. RESULTS: One month after the completion of treatment, 132 of 150 patients (88 percent) in the group assigned to received omeprazole and antibiotics had a negative test for H. pylori, as compared with 7 of 152 (5 percent) in the group assigned to receive omeprazole alone. One year later, dyspepsia had resolved in 33 of 154 patients (21 percent) in the group given omeprazole and antibiotics, as compared with 11 of 154 (7 percent) in the group given omeprazole alone (95 percent confidence interval for the difference, 7 to 22 percent; P<0.001). Among the patients in the group given omeprazole and antibiotics, the symptoms resolved in 26 of the 98 patients (27 percent) who had had symptoms for five years or less, as compared with 7 of the 56 patients (12 percent) who had had symptoms for more than five years (P=0.03). CONCLUSIONS: In patients with H. pylori infection and nonulcer, or functional, dyspepsia, treatment with omeprazole and antibiotics to eradicate the infection is more likely to resolve symptoms than treatment with omeprazole alone. PMID- 9862942 TI - Lack of effect of treating Helicobacter pylori infection in patients with nonulcer dyspepsia. Omeprazole plus Clarithromycin and Amoxicillin Effect One Year after Treatment (OCAY) Study Group. AB - BACKGROUND: It is uncertain whether treatment of Helicobacter pylori infection relieves symptoms in patients with nonulcer, or functional, dyspepsia. METHODS: We conducted a double-blind, multicenter trial of patients with H. pylori infection and dyspeptic symptoms (moderate-to-very-severe pain and discomfort centered in the upper abdomen). Patients were excluded if they had a history of peptic ulcer disease or gastroesophageal reflux disease and had abnormal findings on upper endoscopy. Patients were randomly assigned to seven days of treatment with 20 mg of omeprazole twice daily, 1000 mg of amoxicillin twice daily, and 500 mg of clarithromycin twice daily or with omeprazole alone and then followed up for one year. Treatment success was defined as the absence of dyspeptic symptoms or the presence of minimal symptoms on any of the 7 days preceding the 12-month visit. RESULTS: Twenty of the 348 patients were excluded after randomization because they were not infected with H. pylori, were not treated, or had no data available. For the remaining 328 patients (164 in each group), treatment was successful for 27.4 percent of those assigned to receive omeprazole and antibiotics and 20.7 percent of those assigned to receive omeprazole alone (P=0.17; absolute difference between groups, 6.7 percent; 95 percent confidence interval, -2.6 to 16.0). After 12 months, gastritis had healed in 75.0 percent of the patients in the group given omeprazole and antibiotics and in 3.0 percent of the patients in the omeprazole group (P<0.001); the respective rates of H. pylori eradication were 79 percent and 2 percent. In the group given omeprazole and antibiotics, the rate of treatment success among patients with persistent H. pylori infection was similar to that among patients in whom the infection was eradicated (26 percent vs. 31 percent). There were no significant differences between the groups in the quality of life after treatment. CONCLUSIONS: In patients with nonulcer dyspepsia, the eradication of H. pylori infection is not likely to relieve symptoms. PMID- 9862943 TI - A clinical trial of a chest-pain observation unit for patients with unstable angina. Chest Pain Evaluation in the Emergency Room (CHEER) Investigators. AB - BACKGROUND: Nearly half of patients hospitalized with unstable angina eventually receive a non-cardiac-related diagnosis, yet 5 percent of patients with myocardial infarction are inappropriately discharged from the emergency department. We evaluated the safety, efficacy, and cost of admission to a chest pain observation unit (CPU) located in the emergency department for such patients. METHODS: We performed a community-based, prospective, randomized trial of the safety, efficacy, and cost of admission to a CPU as compared with those of regular hospital admission for patients with unstable angina who were considered to be at intermediate risk for cardiovascular events in the short term. A total of 424 eligible patients were randomly assigned to routine hospital admission (a monitored bed under the care of the cardiology service) or admission to the CPU (where patients were cared for according to a strict protocol including aspirin, heparin, continuous ST-segment monitoring, determination of creatine kinase isoenzyme levels, six hours of observation, and a study of cardiac function). The CPU was managed by the emergency department staff. Patients whose test results were negative were discharged, and the others were hospitalized. Primary outcomes (nonfatal myocardial infarction, death, acute congestive heart failure, stroke, or out-of-hospital cardiac arrest) and use of resources were compared between the two groups. RESULTS: The 212 patients in the hospital-admission group had 15 primary events (13 myocardial infarctions and 2 cases of congestive heart failure), and the 212 patients in the CPU group had 7 events (5 myocardial infarctions, 1 death from cardiovascular causes, and 1 case of congestive heart failure). There was no significant difference in the rate of cardiac events between the two groups (odds ratio for the CPU group as compared with the hospital-admission group, 0.50; 95 percent confidence interval, 0.20 to 1.24). No primary events occurred among the 97 patients who were assigned to the CPU and discharged. Resource use during the first six months was greater among patients assigned to hospital admission than among those assigned to the CPU (P<0.01 by the rank-sum test). CONCLUSIONS: A CPU located in the emergency department can be a safe, effective, and cost-saving means of ensuring that patients with unstable angina who are considered to be at intermediate risk of cardiovascular events receive appropriate care. PMID- 9862944 TI - Atovaquone compared with dapsone for the prevention of Pneumocystis carinii pneumonia in patients with HIV infection who cannot tolerate trimethoprim, sulfonamides, or both. Community Program for Clinical Research on AIDS and the AIDS Clinical Trials Group. AB - BACKGROUND: Although trimethoprim-sulfamethoxazole is the drug of choice for the prevention of Pneumocystis carinii pneumonia, many patients cannot tolerate it and must switch to an alternative agent. METHODS: We conducted a multicenter, open-label, randomized trial comparing daily atovaquone (1500-mg suspension) with daily dapsone (100 mg) for the prevention of P. carinii pneumonia among patients infected with the human immunodeficiency virus who could not tolerate trimethoprim-sulfamethoxazole. The median follow-up period was 27 months. RESULTS: Of 1057 patients enrolled, 298 had a history of P. carinii pneumonia. P. carinii pneumonia developed in 122 of 536 patients assigned to atovaquone (15.7 cases per 100 person-years), as compared with 135 of 521 in the dapsone group (18.4 cases per 100 person-years; relative risk for atovaquone vs. dapsone, 0.85; 95 percent confidence interval, 0.67 to 1.09; P=0.20). The relative risk of death was 1.07 (95 percent confidence interval, 0.89 to 1.30; P=0.45), and the relative risk of discontinuation of the assigned medication because of adverse events was 0.94 (95 percent confidence interval, 0.74 to 1.19; P=0.59). Among the 546 patients who were receiving dapsone at base line, the relative risk of discontinuation because of adverse events was 3.78 for atovaquone as compared with dapsone (95 percent confidence interval, 2.37 to 6.01; P<0.001); among those not receiving dapsone at base line, it was 0.42 (95 percent confidence interval, 0.30 to 0.58; P<0.001). CONCLUSIONS: Among patients who cannot tolerate trimethoprim-sulfamethoxazole, atovaquone and dapsone are similarly effective for the prevention of P. carinii pneumonia. Our results support the continuation of dapsone prophylaxis among patients who are already receiving it. However, among those not receiving dapsone, atovaquone is better tolerated and may be the preferred choice for prophylaxis against P. carinii pneumonia. PMID- 9862945 TI - Images in clinical medicine. "Smiling" alveolar macrophage. PMID- 9862946 TI - The care of HIV-infected adults in the United States. HIV Cost and Services Utilization Study Consortium. AB - BACKGROUND AND METHODS: In order to elucidate the medical care of patients with human immunodeficiency virus (HIV) infection in the United States, we randomly sampled HIV-infected adults receiving medical care in the contiguous United States at a facility other than military, prison, or emergency department facility during the first two months of 1996. We interviewed 76 percent of 4042 patients selected from among the patients receiving care from 145 providers in 28 metropolitan areas and 51 providers in 25 rural areas. RESULTS: During the first two months of 1996, an estimated 231,400 HIV-infected adults (95 percent confidence interval, 162,800 to 300,000) received care. Fifty-nine percent had the acquired immunodeficiency syndrome according to the case definition of the Centers for Disease Control and Prevention, and 91 percent had CD4+ cell counts of less than 500 per cubic millimeter. Eleven percent were 50 years of age or older, 23 percent were women, 33 percent were black, and 49 percent were men who had had sex with men. Forty-six percent had incomes of less than $10,000 per year, 68 percent had public health insurance or no insurance, and 30 percent received care at teaching institutions. The estimated annual direct expenditures for the care of the patients seen during the first two months of 1996 were $5.1 billion; the expenditures for the estimated 335,000 HIV-infected adults seen at least as often as every six months were $6.7 billion, which is about $20,000 per patient per year. CONCLUSIONS: In this national survey we found that most HIV infected adults who were receiving medical care had advanced disease. The patient population was disproportionately male, black, and poor. Many Americans with diagnosed or undiagnosed HIV infection are not receiving medical care at least as often as every six months. The total cost of medical care for HIV-infected Americans accounts for less than 1 percent of all direct personal health expenditures in the United States. PMID- 9862949 TI - Risk adjustment or risk avoidance? PMID- 9862947 TI - Antigen localization and migration in immunity and tolerance. PMID- 9862950 TI - Caring for people with human immunodeficiency virus infection. PMID- 9862951 TI - Helicobacter pylori and nonulcer dyspepsia. PMID- 9862952 TI - The chest-pain unit--ready for prime time? PMID- 9862953 TI - Paying more fairly for Medicare capitated care. PMID- 9862955 TI - RNA binding strategies of ribosomal proteins. AB - Structures of a number of ribosomal proteins have now been determined by crystallography and NMR, though the complete structure of a ribosomal protein rRNA complex has yet to be solved. However, some ribosomal protein structures show strong similarity to well-known families of DNA or RNA binding proteins for which structures in complex with cognate nucleic acids are available. Comparison of the known nucleic acid binding mechanisms of these non-ribosomal proteins with the most highly conserved surfaces of similar ribosomal proteins suggests ways in which the ribosomal proteins may be binding RNA. Three binding motifs, found in four ribosomal proteins so far, are considered here: homeodomain-like alpha helical proteins (L11), OB fold proteins (S1 and S17) and RNP consensus proteins (S6). These comparisons suggest that ribosomal proteins combine a small number of fundamental strategies to develop highly specific RNA recognition sites. PMID- 9862956 TI - Analysis of internal (n-1)mer deletion sequences in synthetic oligodeoxyribonucleotides by hybridization to an immobilized probe array. AB - The purity of a drug substance can influence its toxicity and potency, so impurities must be specifically determined. In the case of synthetic oligodeoxyribonucleotide drugs, however, product complexity makes complete impurity speciation difficult. The goal of the present work was to develop a new analytical method for speciation of individual internal (n-1)mer impurities arising from formal nucleotide deletion in synthetic oligodeoxyribonucleotides. A complete series of oligodeoxyribonucleotide probes were designed, each complementary to an (n-1)mer deletion sequence of the drug in question. Glass plates were used as a solid support for individually immobilizing the entire probe array. The total mixture of internal (n-1) length impurities was isolated from a synthetic oligodeoxyribonucleotide by PAGE and labeled with 35S. Under stringently optimized conditions, only the perfectly sequence-matched oligodeoxyribonucleotide hybridized to each probe, while all other deletion sequences were removed by washing with buffer. The 35S signal intensity of the bound oligodeoxyribonucleotide was proportional to the concentration of each (n 1)mer deletion sequence in the analyte solution. This method has been applied to a number of synthetic phosphorothioate oligodeoxy-ribonucleotide lots and shown to be reliable for speciation and relative quantitation of the internal (n -1)mer deletion sequences present. PMID- 9862957 TI - Identification of promoter elements in mycobacteria: mutational analysis of a highly symmetric dual promoter directing the expression of replication genes of the Mycobacterium plasmid pAL5000. AB - The 120 bp origin of replication (ori) for the Mycobacterium plasmid pAL5000 has been shown to comprise the binding sites for the replication protein RepB as well as the start site of transcription for the repA and repB genes, encoding the replication proteins RepA and RepB. In this work it is demonstrated that a third gene product, Rap, is involved in replication in addition to the previously described proteins. Mycobacterium smegmatis cells transformed with replicons carrying the rap gene recover markedly faster upon electroporation than those transformed with the minimal replicon, which lacks rap. The rap gene, oppositely orientated to repA/B, was shown to be transcribed from a promoter orientated back to-back to and overlapping the repA/B promoter. As a consequence of the extensive dyad symmetry in this region the two promoters share several elements, most of which are situated inside the high-affinity RepB-binding motif in the ori. Transcription of rap runs through the low-affinity RepB-binding site, which is part of the ori and necessary for replication. Both promoters were shown to be repressed by RepB. These divergent promoters were studied through site-specific mutagenesis in a xylE reporter gene assay. The analysis furnished evidence supporting the existence of a distal as well as a proximal element in mycobacterial promoters. PMID- 9862958 TI - Detailed analysis of stem I and its 5' and 3' neighbor regions in the trans acting HDV ribozyme. AB - To determine the stem I structure of the human hepatitis delta virus (HDV) ribozyme, which is related to the substrate sequence in the trans -acting system, we kinetically studied stem I length and sequences. Stem I extension from 7 to 8 or 9 bp caused a loss of activity and a low amount of active complex with 9 bp in the trans -acting system. In a previous report, we presented cleavage in a 6 bp stem I. The observed reaction rates indicate that the original 7 bp stem I is in the most favorable location for catalytic reaction among the possible 6-8 bp stems. To test base specificity, we replaced the original GC-rich sequence in stem I with AU-rich sequences containing six AU or UA base pairs with the natural +1G.U wobble base pair at the cleavage site. The cis -acting AU-rich molecules demonstrated similar catalytic activity to that of the wild-type. In trans acting molecules, due to stem I instability, reaction efficiency strongly depended on the concentration of the ribozyme-substrate complex and reaction temperature. Multiple turnover was observed at 37 degreesC, strongly suggesting that stem I has no base specificity and more efficient activity can be expected under multiple turnover conditions by substituting several UA or AU base pairs into stem I. We also studied the substrate damaging sequences linked to both ends of stem I for its development in therapeutic applications and confirmed the functions of the unique structure. PMID- 9862959 TI - Exogenous expression of a dominant negative RORalpha1 vector in muscle cells impairs differentiation: RORalpha1 directly interacts with p300 and myoD. AB - ROR/RZR is an orphan nuclear receptor that has no known ligand in the 'classical sense'. In the present study we demonstrate that RORalpha is constitutively expressed during the differentiation of proliferating myoblasts to post-mitotic multinucleated myotubes, that have acquired a contractile phenotype. Exogenous expression of dominant negative RORalpha1DeltaE mRNA in myogenic cells significantly reduces the endogenous expression of RORalpha1 mRNA, represses the accumu-lation and delays the activation of mRNAs encoding MyoD and myogenin [the muscle-specific basic helix-loop-helix (bHLH) proteins] and p21(Waf-1/Cip-1) (a cdk inhibitor). Immunohistochemistry demonstrates that morpho-logical differentiation is delayed in cells expressing the RORDeltaE transcript. Furthermore, the size and development of mutlinucleated myotubes is impaired. The E region of RORalpha1 interacts with p300, a cofactor that functions as a coactivator in nuclear receptor and MyoD-mediated transactivation. Consistent with the functional role of RORalpha1 in myogenesis, we observed that RORalpha1 directly interacts with the bHLH protein MyoD. This interaction was mediated by the N-terminal activation domain of the bHLH protein, MyoD, and the RORalpha1 DNA binding domain/C region. Furthermore, we demonstrated that p300, RORalpha1 and MyoD interact in a non-competitive manner. In conclusion, this study provides evidence for a biological role and positive influence of RORalpha1 in the cascade of events involved in the activation of myogenic-specific markers and cell cycle regulators and suggests that crosstalk between theretinoid-relatedorphan (ROR) nuclear receptors and the myogenic bHLH proteins has functional consequences for differentiation. PMID- 9862961 TI - A vector based on the SV40 origin of replication and chromosomal S/MARs replicates episomally in CHO cells. AB - We have developed an episomal replicating expression vector in which the SV40 gene coding for the large T-antigen was replaced by chromosomal scaffold/matrix attached regions. Southern analysis as well as vector rescue experiments in CHO cells and in Escherichia coli demonstrate that the vector replicates episomally in CHO cells. It occurs in a very low copy number in the cells and is stably maintained over more than 100 generations without selection pressure. PMID- 9862960 TI - Immunological analysis of potato leafroll luteovirus (PLRV) P1 expression identifies a 25 kDa RNA-binding protein derived via P1 processing. AB - Mono- and polyclonal antibodies directed against different domains of the potato leafroll luteovirus (PLRV) P1 (ORF1) protein were applied to the analysis of P1 expression during PLRV replication in planta. Western analyses detected P1 and a protein of approximately 25 kDa (P1-C25) that accumulated to readily detectable amounts in PLRV-infected plants, but was not detected by in vitro cell-free translation of P1. P1-C25 represents the C-terminus of P1 and is a proteolytic cleavage product produced during P1 processing. On the basis of its molecular weight, the N-terminus of P1-C25 is either identical to or located adjacent to the previously identified PLRV genome-linked protein, VPg. P1-C25 is not associated with virus particles, and subcellular localization experiments detected P1-C25, but not P1, in the membrane and cytoplasmic fractions of PLRV infected cells. In addition, P1-C25 exhibits nucleic acid-binding properties. On the basis of its biosynthesis, localization and biochemical properties, P1-C25 may facilitate the formation of P1/PLRV RNA complexes in which the spatial proximity allows for covalent bond formation between PLRV RNA and VPg. PMID- 9862962 TI - Identification of a stem-loop structure important for polyadenylation at the murine IgM secretory poly(A) site. AB - We have previously shown that a distal GU-rich downstream element of the mouse IgM secretory poly(A) site is important for polyadenylation in vivo and for polyadenylation specific complex formation in vitro. This element can be predicted to form a stem-loop structure with two asymmetric internal loops. As stem-loop structures commonly define protein RNA binding sites, we have probed the biological activity of the secondary structure of this element. We show that mutations affecting the stem of the structure abolish the biological activity of this element in vivo and in vitro at the level of cleavage and polyadenylation specificity factor/cleavage stimulation factor complex formation and that both internal loops contribute to the enhancing effect of the sequence in vivo. Lead (II) cleavage patterns and RNase H probing of the sequence element in vitro are consistent with the predicted secondary structure. Furthermore, mobility on native PAGE suggests a bent structure. We propose that the secondary structure of this downstream element optimizes its interaction with components of the polyadenylation complex. PMID- 9862963 TI - Alternative function of a protein kinase homology domain in 2', 5'-oligoadenylate dependent RNase L. AB - RNase L is the 2',5'-oligoadenylate (2-5A)-dependent endoribonuclease that functions in interferon action and apoptosis. One of the intriguing, albeit unexplained, features of RNase L is its significant homology to protein kinases. Despite the homology, however, no protein kinase activity was detected during activation and RNA cleavage reactions with human RNase L. Similarly, the kinase plus ribonuclease domains of RNase L produced no detectable protein kinase activity in contrast to the phosphorylation obtained with homologous domains of the related kinase and endoribonuclease, yeast IRE1p. In addition, neither ATP nor pA(2'p5'A)3was hydrolyzed by RNase L. To further investigate the function of the kinase homology in RNase L, the conserved lysine at residue 392 in protein kinase-like domain II was replaced with an arginine residue. The resulting mutant, RNase LK392R, showed >100-fold decreases in 2-5A-dependent ribonuclease activity without reducing 2-5A- or RNA-binding activities. The greatly reduced activity of RNase LK392Rwas correlated to a defect in the ability of RNase L to dimerize. These results demonstrate a critical role for lysine 392 in the activation and dimerization of RNase L, thus suggesting that these two activities are intimately linked. PMID- 9862964 TI - The ability of the HIV-1 AAUAAA signal to bind polyadenylation factors is controlled by local RNA structure. AB - The 5' and 3' ends of HIV-1 transcripts are identical in sequence. This repeat region (R) folds a stem-loop structure that is termed the poly(A) hairpin because it contains polyadenylation or poly(A) signals: the AAUAAA hexamer motif, the cleavage site and part of the GU-rich downstream element. Obviously, HIV-1 gene expression necessitates differential regulation of the two poly(A) sites. Previous transfection experiments indicated that the wild-type poly(A) hairpin is slightly inhibitory to the process of polyadenylation, and further stabilization of the hairpin inhibited polyadenylation completely. In this study, we tested wild-type and mutant transcripts with poly(A) hairpin structures of differing thermodynamic stabilities for the in vitro binding of polyadenylation factors. Mutant transcripts with a destabilized hairpin efficiently bound the polyadenylation factors, which were provided either as purified proteins or as nuclear extract. The RNA mutant with a stabilized hairpin did not form this 'poly(A) complex'. Additional mutations that repair the stability of this hairpin restored the binding capacity. Thus, an inverse correlation was measured between the stability of the poly(A) hairpin and its ability to interact with polyadenylation factors. The wild-type HIV-1 transcript bound the polyadenylation factors suboptimally, but full activity was obtained in the presence of the USE enhancer element that is uniquely present upstream of the 3' poly(A) site. We also found that sequences of the HIV-1 leader, which are uniquely present downstream of the 5' poly(A) site, inhibit formation of the poly(A) complex. This inhibition could not be ascribed to a specific leader sequence, as we measured a gradual loss of complex formation with increasing leader length. We will discuss the regulatory role of RNA structure and the repressive effect of leader sequences in the context of differential HIV-1 polyadenylation. PMID- 9862965 TI - Pogo transposase contains a putative helix-turn-helix DNA binding domain that recognises a 12 bp sequence within the terminal inverted repeats. AB - Pogo is a transposable element with short terminal inverted repeats. It contains two open reading frames that are joined by splicing and code for the putative pogo transposase, the sequence of which indicates that it is related to the transposases of members of the Tc1/mariner family as well as proteins that have no known transposase activity including the centromere binding protein CENP-B. We have shown that the N-terminal region of pogo transposase binds in a sequence specific manner to the ends of pogo and have identified residues essential for this. The results are consistent with a prediction that DNA binding is due to a helix-turn-helix motif within this region. The transposase recognises a 12 bp sequence, two copies of which are present at each end of pogo DNA. The outer two copies occur as inverted repeats 14 nucleotides from each end of the element, and contain a single base mismatch and indicate the inverted repeats of pogo are 26 nucleotides long. The inner copies occur as direct repeats, also with a single mismatch. PMID- 9862966 TI - A key role for replication factor C in DNA replication checkpoint function in fission yeast. AB - Replication factor C (RF-C) is a five subunit DNA polymerase (Pol) delta/straightepsilon accessory factor required at the replication fork for loading the essential processivity factor PCNA onto the 3'-ends of nascent DNA strands. Here we describe the genetic analysis of the rfc2 +gene of the fission yeast Schizosaccharomyces pombe encoding a structural homologue of the budding yeast Rfc2p and human hRFC37 proteins. Deletion of the rfc2 + gene from the chromosome is lethal but does not result in the checkpoint-dependent cell cycle arrest seen in cells deleted for the gene encoding PCNA or for those genes encoding subunits of either Pol delta or Pol straightepsilon. Instead, rfc2 Delta cells proceed into mitosis with incompletely replicated DNA, indicating that the DNA replication checkpoint is inactive under these conditions. Taken together with recent results, these observations suggest a simple model in which assembly of the RF-C complex onto the 3'-end of the nascent RNA-DNA primer is the last step required for the establishment of a checkpoint-competent state. PMID- 9862967 TI - RAV1, a novel DNA-binding protein, binds to bipartite recognition sequence through two distinct DNA-binding domains uniquely found in higher plants. AB - We have cloned and characterized two novel DNA binding proteins designated RAV1 and RAV2 from Arabidopsis thaliana. RAV1 and RAV2 contain two distinct amino acid sequence domains found only in higher plant species. The N-terminal regions of RAV1 and RAV2 are homologous to the AP2 DNA-binding domain present in a family of transcription factors represented by the Arabidopsis APETALA2 and tobacco EREBP proteins, while the C-terminal region exhibits homology to the highly conserved C terminal domain, designated B3, of VP1/ABI3 transcription factors. Binding site selection assays using a recombinant glutathione S-transferase fusion protein have revealed that RAV1 binds specifically to bipartite recognition sequences composed of two unrelated motifs, 5'-CAACA-3' and 5'-CACCTG-3', separated by various spacings in two different relative orientations. Analyses using various deletion derivatives of the RAV1 fusion protein show that the AP2 and B3-like domains of RAV1 bind autonomously to the CAACA and CACCTG motifs, respectively, and together achieve a high affinity and specificity of binding. From these results, we suggest that the AP2 and B3-like domains of RAV1 are connected by a highly flexible structure enabling the two domains to bind to the CAACA and CACCTG motifs in various spacings and orientations. PMID- 9862968 TI - A re-investigation of the thio effect at the hammerhead cleavage site. AB - The effect of introducing a phosphorothioate at the hammerhead cleavage site was investigated using a kinetically well-characterized hammerhead. In buffers containing Mg ion, the RP-phosphorothioate isomer cleaved 2000- to 80 000-fold slower than the SPisomer or the unmodified RNA substrate. Addition of low concentrations of several thiophilic metal ions, especially Cd2+, to these reactions is sufficient to fully restore the cleavage rate of the RPsubstrate without affecting cleavage rate of the all-oxygen or SPsubstrate. Thus, a model proposing coordination of a divalent metal ion to the pro-R oxygen at the hammerhead cleavage site appears justified. PMID- 9862969 TI - Structural requirements of the higher order RNA kissing element in the enteroviral 3'UTR. AB - The origin of replication ( oriR ) involved in the initiation of (-) strand enterovirus RNA synthesis is a quasi-globular multi-domain RNA structure which is maintained by a tertiary kissing interaction. The kissing interaction is formed by base pairing of complementary sequences within the predominant hairpin-loop structures of the enteroviral 3' untranslated region. In this report, we have fully characterised the kissing interaction. Site-directed mutations which affected the different base pairs involved in the kissing interaction were generated in an infectious coxsackie B3 virus cDNA clone. The kissing interaction appeared to consist of 6 bp. Distortion of the interaction by mispairing of each of the base pairs involved in this higher order RNA structure resulted in either temperature sensitive or lethal phenotypes. The nucleotide constitution of the base which gaps the major groove of the kissing domain was not relevant for virus growth. The reciprocal exchange of the complete sequence involved in the kissing resulted in a mutant virus with wild type virus growth characteristics arguing that the base pair constitution is of less importance for the initiation of (-) strand RNA synthesis than the existence of the tertiary structure itself. PMID- 9862970 TI - Increased levels of E2F-1-dependent DNA binding activity after UV- or gamma irradiation. AB - In mammalian cells, DNA damage induces robust changes in gene expression and these changes contribute to the proper execution of cellular responses to DNA damage, including DNA repair, cell cycle arrest and apoptosis. The transcription factor E2F-1 has been suggested to play a key role in the regulation of cell cycle-dependent gene expression and apoptosis. These activities depend on the ability of E2F-1 to form functionally active DNA binding complexes. Here we describe an assay that allows one to measure E2F-1 DNA binding activity in naive cells. We find that DNA damage, generated by UV- or gamma-irradiation, prompts increased production of E2F-1 DNA binding activity, which, at least in part, originates from alterations in E2F-1 protein levels. These findings represent an indication for a role of the transcription factor E2F-1 in the DNA damage response pathway. PMID- 9862971 TI - Insertion of dGMP and dAMP during in vitro DNA synthesis opposite an oxidized form of 7,8-dihydro-8-oxoguanine. AB - Oxidative damage to DNA bases commonly resultsin the formation of oxidized purines, particularly 7,8-dihydro-8-oxoguanine (8-oxoG) and 7,8-dihydro-8 oxoadenine (8-oxoA), the former being a well-known mutagenic lesion. Since 8-oxoG is readily subject to further oxidation compared with normal bases, the insertion of a base during DNA synthesis opposite an oxidized form of 8-oxoG was investigated in vitro. A synthetic template containing a single 8-oxoG lesion was first treated with different one-electron oxidants or under singlet oxygen conditions and then subjected to primer extension catalyzed by Klenow fragment exo- (Kf exo-), calf thymus DNA polymerase alpha (pol alpha) or human DNA polymerase beta (pol beta). Consistent with previous reports, dAMP and dCMP are inserted selectively opposite 8-oxoG with all three DNA polymerases. Interestingly, oxidation of 8-oxoG was found to induce dAMP and dGMP insertion opposite the lesion by Kf exo- with transient inhibition of primer extension occurring at the site of the modified base. Furthermore, the lesion constitutes a block during DNA synthesis by pol alpha and pol beta. Experiments with an 8-oxoA modified template oligonucleotide show that both 8-oxoA and an oxidized form of 8 oxoA direct insertion of dTMP by Kf exo-. Mass spectrometric analysis of 8-oxoG containing oligonucleotides before and after oxidation with IrCl62-are consistent with oxidation of primarily the 8-oxoG site, resulting in formation of a guanidinohydantoin moiety as the major product. No evidence for formation of abasic sites was obtained. These results demonstrate that an oxidized form of 8 oxoG, possibly guanidinohydantoin, may direct misreading and misinsertion of dNTPs during DNA synthesis. If such a process occurred in vivo, it would represent a point mutagenic lesion leading to G-->T and G-->C transversions. However, the corresponding oxidized form of 8-oxoA primarily shows correct insertion of T during DNA synthesis with Kf exo-. PMID- 9862972 TI - Role of the conserved lysine 80 in stabilisation of NF-kappaB p50 DNA binding. AB - The transcriptional rate of a variety of genes involved in acute-phase response, inflammation, lymphocytic activation, and cell growth or differentiation, is regulated by the DNA binding activity of the inducible transcription factor NF kappaB. NF-kappaB p50 homodimers bind specifically to DNA, via base and backbone contacts mediated by residues in the flexible loops which link secondary structure elements in both of its two distinct domains. However, it has been suggested that additional contacts which stabilise DNA binding are made by lysine residues located in the C-terminus of the flexible loop which connects A and B beta-sheets of the N-terminal domain of p50. To determine the importance of each of the lysine residues in this region (K77, K79, K80), a series of mutated p50 proteins were generated in which the lysines were changed to alanines. The DNA binding properties of these mutants were analysed by gel electrophoresis DNA binding assays and surface plasmon resonance. This study revealed that the C terminus of AB loop interacts with DNA through an additional lysine-phosphate backbone ionic bond which makes a significant contribution to the binding energy, thus stabilising the complex. The lysine residue responsible for this interaction is K80 which is conserved in all NF-kappaB/Rel/Dorsal molecules. PMID- 9862973 TI - Identification of three conserved regions in the DREF transcription factors from Drosophila melanogaster and Drosophila virilis. AB - The genes for a DNA replication-related element-binding factor (DREF) were isolated from Drosophila melanogaster and Drosophila virilis, and their nucleotide sequences were determined. Drosophila virilis DREF consists of 742 amino acid residues, which is 33 amino acids longer than D.melanogaster DREF. Comparison of the amino acid sequences revealed that D.virilis DREF is 71% identical to its D. melanogaster homolog. Three highly conserved regions were identified at amino acid positions 14-182 (CR1), 432-568 (CR2) and 636-730 (CR3) of the D.virilis DREF, with 86.4, 86.1 and 83.3% identities, respectively. Transgenic flies in which expression of three conserved regions of D.melanogaster DREF was targeted to the eye imaginal disc were established. Expression of CR1 in the developing eye imaginal discs resulted in a severe rough eye phenotype in adult flies. Expression of CR3 also caused a rough eye phenotype, while that of CR2 had no apparent effect on eye morphology. Expression of either CR1 or CR3 in eye imaginal disc cells inhibited cell cycle progression and reduced incorporation of 5-bromo-2'-deoxyuridine into the S-phase zone (the second mitotic wave) behind the morphogenetic furrow. The results indicate that both CR1 and CR3 are important for DREF functions. PMID- 9862975 TI - Transcription and RNA processing of mammalian genes in Saccharomyces cerevisiae. AB - The recognition of mammalian genes encoded within a mouse yeast artificial chromosome (YAC) by the yeast transcription and RNA processing machinery was investigated. Transcripts from five genes known to be encoded by the YAC were all found in the total yeast RNA. Of 12 mouse introns assayed, six were correctly spliced by the yeast. However, an abnormal transcription of mouse DNA was also observed. Three genes of three tested were transcribed both from their sense and antisense strands and all tested microsatellite, inter-repetitive and anonymous mouse loci were detected in the YAC clone RNA. An RNA transcript from a well defined intergenic region of two head-to-head oriented mouse genes was detected by RT-PCR and by RNase protection assay. These results indicate the presence of multiple yeast-specific transcription sites in the mouse DNA. 3' RACE experiments demonstrated the inability of the yeast to use the mouse polyadenylation signals. Thus, a method for isolation of mammalian exons based on a YAC clone RNA is likely to produce a high background, because the enrichment with mammalian exons in the YAC RNA is low. Nevertheless, YAC clones can serve as in vivo test tubes to study the conservation of RNA processing sequences. PMID- 9862974 TI - Increased nuclear factor 1 binding to its nucleosomal site mediated by sequence dependent DNA structure. AB - The organization of DNA into chromatin is important in the regulation of transcription, by influencing the access of transcription factors to their DNA binding sites. Nuclear factor 1 (NF-1) is a transcription factor which binds to DNA constitutively and which interacts with its cognate DNA site with high affinity. However, this affinity is drastically reduced, approximately 100- to 300-fold, when the binding site is organized into a nucleosome. Here we demonstrate that the introduction of stretches of adenines of length 5 nt (A tracts) on both sides of the NF-1 binding site has a distinct effect on NF-1 binding to a nucleosomal, but not to a free, NF-1 binding site. The position of the A-tracts, relative to the rotational phase of a synthetic DNA bending sequence, the TG-motif, decides whether the NF-1 affinity increases or decreases. The NF-1 binding affinity is seven times stronger when the flanking A-tracts are positioned out-of-phase with the TG-motif than it is when the A-tracts are positioned in-phase with the TG-motif. We demonstrate that this effect correlates with differences in DNA curvature and apparent histone octamer affinity. We conclude that DNA curvature influences the local histone-DNA contacts and hence the accessibility of the NF-1 site in a nucleosome context. PMID- 9862976 TI - Histone stoichiometry and DNA circularization in archaeal nucleosomes. AB - Recombinant (r)HMfB (archaealhistone B fromMethanothermusfervidus) formed complexes with increasing stability with DNA molecules increasing in length from 52 to 100 bp, but not with a 39 bp molecule. By using125I-labeled rHMfB-YY (an rHMfB variant with I31Y and M35Y replacements) and32P-labeled 100 bp DNA, these complexes, designated archaeal nucleosomes, have been shown to contain an archaeal histone tetramer. Consistent with DNA bending and wrapping, addition of DNA ligase to archaeal nucleosomes assembled with 88 and 128 bp DNAs resulted in covalently-closed monomeric circular DNAs which, following histone removal, were positively supercoiled based on their electrophoretic mobilities in the presence of ethidium bromide before and after relaxation by calf thymus topoisomerase I. Ligase addition to mixtures of rHMfB with 53 or 30 bp DNA molecules also resulted in circular DNAs but these were circular dimers and trimers. These short DNA molecules apparently had to be ligated into longer linear multimers for assembly into archaeal nucleosomes and ligation into circles. rHMfB assembled into archaeal nucleosomes at lower histone to DNA ratios with the supercoiled, circular ligation product than with the original 88 bp linear version of this molecule. Archaeal histones are most similar to the globular histone fold region of eukaryal histone H4, and the results reported are consistent with archaeal nucleosomes resembling the structure formed by eukaryal histone (H3+H4)2tetramers. PMID- 9862977 TI - A DNA polymerase stop assay for G-quadruplex-interactive compounds. AB - We have developed and characterized an assay for G-quadruplex-interactive compounds that makes use of the fact that G-rich DNA templates present obstacles to DNA synthesis by DNA polymerases. Using Taq DNA polymerase and the G quadruplex binding 2, 6-diamidoanthraquinone BSU-1051, we find that BSU-1051 leads to enhanced arrest of DNA synthesis in the presence of K+by stabilizing an intramolecular G-quadruplex structure formed by four repeats of either TTGGGG or TTAGGG in the template strand. The data provide additional evidence that BSU-1051 modulates telomerase activity by stabilization of telomeric G-quadruplex DNA and point to a polymerase arrest assay as a sensitive method for screening for G quadruplex-interactive agents with potential clinical utility. PMID- 9862978 TI - The intrinsic structure and stability of out-of-alternation base pairs in Z-DNA. AB - Alternating pyrimidine-purine sequences typically form Z-DNA, with the pyrimidines in the anti and purines in the syn conformations. The observation that dC and dT nucleotides can also adopt the syn conformation (i.e. the nucleotides are out-of-alternation) extends the range of sequences that can convert to this left-handed form of DNA. Here, we study the effects of placing two adjacent d(G*C) base pairs as opposed to a single d(G*C) base pair or two d(A*T) base pairs out-of-alternation by comparing the structure of d(m5CGGCm5CG)2with the previously published structures of d(m5CGGGm5CG)*d(m5CGCCm5CG) and d(m5CGATm5CG)2. A high buckle and loss of stacking interactions are observed as intrinsic properties of the out-of alternation base pairs regardless of sequence and the context of the dinucleotide. From solution titrations, we find that the destabilizing effect of out-of-alternation d(G*C) base pairs are identical whether these base pairs are adjacent or isolated. We can therefore conclude that it is these intrinsic distortions in the structure of the base pairs and not neighboring effects that account for the inability of out-of-alternation base pairs to adopt the left handed Z conformation. PMID- 9862979 TI - Inhibition of a DNA-helicase by peptide nucleic acids. AB - Bis-peptide nucleic acid (bis-PNA) binding results in D-loop formation by strand displacement at complementary homopurine stretches in DNA duplexes. Transcription and replication in intact cells is mediated by multienzymatic complexes involving several proteins other than polymerases. The behaviour of the highly stable clamp structure formed by bis-PNAs has thus far been studied with respect to their capacity to arrest RNA polymerases. Little attention has been given to their recognition and processing by DNA helicases. In this report we have investigated the inhibitory effect of a bis-PNA on the DNA-helicase activity of the well characterized herpes simplex type I UL9 protein. Unwinding by UL9 of a synthetic substrate is significantly inhibited by a bis-PNA and the addition of the ICP8 protein, which increases UL9 processivity, does not relieve this inhibition. PMID- 9862980 TI - The crystal structure of the RNA/DNA hybrid r(GAAGAGAAGC). d(GCTTCTCTTC) shows significant differences to that found in solution. AB - The crystal structure of the RNA/DNA hybrid r(GAAGAGAAGC). d(GCTTCTCTTC) has been solved and refined at 2.5 A resolution. The refinement procedure converged at R = 0.181 for all reflections in the range 20.0-2.5 A. In the crystal, the RNA/DNA hybrid duplex has an A' conformation with all but one of the nucleotide sugar moieties adopting a C3'- endo (N) conformation. Both strands in the double helix adopt a global conformation close to the A-form and the width of the minor groove is typical of that found in the crystal structures of other A-form duplexes. However, differences are observed between the RNA and DNA strands that make up the hybrid at the local level. In the central portion of the duplex, the RNA strand has backbone alpha, beta and gamma torsion angles that alternate between the normal gauche -/ trans / gauche + conformation and an unusual trans / trans / trans conformation. Coupled with this so-called 'alpha/gamma flipping' of the backbone torsion angles, the distance between adjacent phosphorous atoms on the RNA strand systematically varies. Neither of these phenomena are observed on the DNA strand. The structure of the RNA/DNA hybrid presented here differs significantly from that found in solution for this and other sequences. Possible reasons for these differences and their implications for the current model of RNase H activity are discussed. PMID- 9862981 TI - Structure and function of a small RNA that selectively inhibits internal ribosome entry site-mediated translation. AB - A 60 nt long RNA termed IRNA, isolated from the yeast Saccharomyces cerevesiae, was previously shown to selectively block internal ribosome entry site (IRES) mediated translation without interfering with cap-dependent translation of cellular mRNAs both in vivo and in vitro. IRNA specifically bound cellular proteins believed to be important for IRES-mediated translation. We demonstrate here that a complementary copy of IRNA (cIRNA) is also active in blocking IRES mediated translation and that it binds many of the same cellular proteins that IRNA does. We have probed the secondary structure of both IRNA and cIRNA using single-strand- and double-strand-specific nucleases as well as using oligonucleotide hybridization followed by RNase H digestion. Both IRNA and cIRNA share secondary structural homology, although distinct differences do exist between the two structures. Mutational analysis of IRNA shows that sequences that form both the main stem and one loop are critical for its translation inhibitory activity. Maintenance of the established secondary structure appears to be required for both IRNA's ability to bind cellular trans -acting proteins believed to be required for IRES-mediated translation and its ability to block IRES mediated translation. PMID- 9862982 TI - Tandem repeats finder: a program to analyze DNA sequences. AB - A tandem repeat in DNA is two or more contiguous, approximate copies of a pattern of nucleotides. Tandem repeats have been shown to cause human disease, may play a variety of regulatory and evolutionary roles and are important laboratory and analytic tools. Extensive knowledge about pattern size, copy number, mutational history, etc. for tandem repeats has been limited by the inability to easily detect them in genomic sequence data. In this paper, we present a new algorithm for finding tandem repeats which works without the need to specify either the pattern or pattern size. We model tandem repeats by percent identity and frequency of indels between adjacent pattern copies and use statistically based recognition criteria. We demonstrate the algorithm's speed and its ability to detect tandem repeats that have undergone extensive mutational change by analyzing four sequences: the human frataxin gene, the human beta T cellreceptor locus sequence and two yeast chromosomes. These sequences range in size from 3 kb up to 700 kb. A World Wide Web server interface atc3.biomath.mssm.edu/trf.html has been established for automated use of the program. PMID- 9862983 TI - Dimerization of the guanine-adenine repeat strands of DNA. AB - Jovin and co-workers have demonstrated that DNA strands containing guanine adenine repeats generate a parallel-stranded homoduplex. Here we propose that the homoduplex is a dimer of the ordered single strand discovered by Fresco and co workers at acid pH. The Fresco single strand is shown here to be stabilized in aqueous ethanol where adenine is not protonated. Furthermore, we demonstrate that the strands dimerize at higher salt concentrations without significantly changing their conformation, so that the dimerization is non-cooperative. Hence, the Jovin homoduplex can form through a non-cooperative dimerization of two cooperatively melting single strands. The available data indicate that the guanines stabilize the Fresco single strand whereas the adenines cause dimerization owing to their known intercalation or clustering tendency. The guanine-adenine repeat dimer seems to be a DNA analog of the leucine zipper causing dimerization of proteins. PMID- 9862984 TI - U1 snRNA is cleaved by RNase III and processed through an Sm site-dependent pathway. AB - Core snRNP proteins bind snRNA through the conserved Sm site, PuA(U)n>/=3GPu. While yeast U1 snRNA has three matches to the Sm consensus, the U1 3'-terminal Sm site was found to be both necessary and sufficient for U1 function. Mutation of this site inhibited pre-mRNA splicing, blocked cell division and resulted in the accumulation of two 3'-extended forms of the U1 snRNA. Cells which harbor the Sm site mutation lack mature U1 RNA (U1alpha) but have a minor polyadenylated species, U1gamma, and a prominent, non-polyadenylated species, U1beta. Metabolic depletion of the essential Sm core protein, Smd1p, also resulted in the increased accumulation of U1beta and U1gamma. In vitro, synthetic U1 precursors were cleaved by Rnt1p (RNase III) very near the U1beta 3'-end observed in vivo. We propose that U1beta is an Rnt1p-cleaved intermediate and that U1 maturation to the U1alpha form occurs through an Sm-sensitive step. Interestingly, both U1alpha and a second, much longer RNA, U1straightepsilon, were produced in an rnt1 mutant strain. These results suggest that yeast U1 snRNA processing may progress through Rnt1p-dependent and Rnt1p-independent pathways, both of which require a fun ctional Sm site for final snRNA maturation. PMID- 9862985 TI - Conserved sequence preference in DNA binding among recombination proteins: an effect of ssDNA secondary structure. AB - Repetitive sequences have been proposed to be recombinogenic elements in eukaryotic chromosomes. We tested whether dinucleotide repeats sequences are preferential sites for recombination because of their high affinity for recombination enzymes. We compared the kinetics of the binding of the scRad51, hsRad51 and ecRecA proteins to oligonucleotides with repeats of dinucleotides GT, CA, CT, GA, GC or AT. Since secondary structures in single-stranded DNA (ssDNA) act as a barrier to complete binding we measured whether these oligonucleotides are able to form stable secondary structures. We show that the preferential binding of recombination proteins is conserved among the three proteins and is influenced mainly by secondary structures in ssDNA. PMID- 9862986 TI - 'LABNOTE', a laboratory notebook system designed for academic genomics groups. AB - We have developed a relational laboratory database system, adapted to the daily book-keeping needs of laboratories that must keep track of information acquired on hundreds or thousands of clones in an effective and user-friendly fashion. Data, whether final or related to experiments in progress, can be accessed in many different ways, e.g. by clone name, by gene, by experiment or through DNA sequence. Updating, import and export of results is made easier by specially developed tools. This system, in network version, serves several groups in our Institute and (over the Internet) elsewhere, and is instrumental in collaborative studies based on expression profiling. It can be used in many similar situations involving progressiveaccumulation of information on sets of clones or related objects. PMID- 9862987 TI - AP site structural determinants for Fpg specific recognition. AB - The binding of Escherichia coli and Lactococcus lactis Fapy-DNA glyosylase (Fpg) proteins to DNA containing either cyclic or non-cyclic abasic (AP) site analogs was investigated by electrophoretic mobility shift assay (EMSA) and by footprinting experiments. We showed that the reduced AP site is the best substrate analog for the E.coli and L.lactis enzymes ( K Dapp = 0.26 and 0.5 nM, respectively) as compared with the other analogs tested in this study ( K Dapp >2.8 nM). The 1,3-propanediol (Pr) residue-containing DNA seems to be the minimal AP site structure allowing a Fpg specific DNA binding, since the ethyleneglycol residue is not specifically bound by these enzymes. The newly described cyclopentanol residue is better recognized than tetrahydrofuran (for the E.coli Fpg, K Dapp = 2.9 and 25 nM, respectively). These results suggest that the hemiacetal form of the AP site is negatively discriminated by the Fpg protein suggesting a hydrogen bond between the C4'-hydroxyl group of the sugar and a Fpg residue. High-resolution hydroxyl radical footprinting using a duplex containing Pr shows that Fpg binds to six nucleotides on the strand containing the AP site and only the base opposite the lesion on the undamaged complementary strand. This comparative study provides new information about the molecular mechanism involved in the Fpg AP lyase activity. PMID- 9862989 TI - Chemical ligation of oligodeoxyribonucleotides on circular DNA templates. AB - We report the use of small circular DNA as a triplex-directing template for the highly efficient chemical ligation of oligodeoxyribonucleotides (ODNs) using cyanogen bromide (BrCN). These investigations compared the use of a linear homopyrimidine DNA template (17mer) and a circular pyrimidine-rich DNA template (44mer) for directing the chemical ligation of two homopurine ODNs (6mer + 11mer). The effects of substrate/template ratio, buffer, salt, ionic strength, pH and temperature have been examined in the BrCN activated ligation reactions. The optimal yield of 51% for ligation on the linear template was at pH 6.0, 200 mM MgCl2, 4 degreesC. In contrast, near quantitative ligation on the circular template occurred at higher pH, higher temperature, and showed less dependence on Mg2+concentration (97% yield, pH 7.5, 200 mM MgCl2, 25 degreesC). The relative observed rate of the ligation reaction was a minimum of 35 times faster on the circular DNA template relative to the linear template at pH 7.5, 200 mM MgCl2, 4 degreesC. These investigations reveal that chemical ligation of short ODNs on circularized DNA templates through triplex formation is a highly efficient process over a broad range of conditions. PMID- 9862988 TI - Nucleotide excision repair affects the stability of long transcribed (CTG*CAG) tracts in an orientation-dependent manner in Escherichia coli. AB - The influence of nucleotide excision repair (NER), the principal in vivo repair system for DNA damages, was investigated in Escherichia coli with uvrA, uvrB and uvrAuvrB mutants with the triplet repeat sequences (TRS) involved in myotonic dystrophy, the fragile X syndrome and Friedreich's ataxia. (CTG*CAG)175was more stable when the (CTG) strand was transcribed than when the (CAG) strand was transcribed in the alternate orientation. A lack of the UvrA protein dramatically increases the instability of this TRS in vivo as compared with the stability of the same sequence in uvrB mutant, which produces an intact UvrA protein. We propose that transcription transiently dissociates the triplet repeat complementary strands enabling the non-transcribed strand to fold into a hairpin conformation which is then sufficiently stable that replication bypasses the hairpin to give large deletions. If the TRS was not transcribed, fewer deletions were observed. Alternatively, in the uvrA-mutant, the hairpins existing on the lagging strand will suffer bypass DNA synthesis to generate deleted molecules. Hence, NER, functionally similar in both prokaryotes and eukaryotes, is an important factor in the genetic instabilities of long transcribed TRS implicated in human hereditary neuro-logical diseases. PMID- 9862990 TI - The DEAD box RNA helicase family in Arabidopsis thaliana. AB - The numerous genomic sequences and ESTs released by the Arabidopsis thaliana Genome Initiative (AGI) have allowed a systematic and functional study of the DEAD box RNA helicase family. Sequencing and in silico analysis led to the characterization of 28 novel A. thaliana DEAD box RNA helicases forming a family of 32 members, named AtRH. Fourteen AtRH genes with an unexpected heterogeneous mosaic structure are described and compared bringing new information about the genesis of the gene family. The mapping of the AtRH genes shows their repartition on the five chromosomes without clustering and therefore AtRH s have been estimated to 60 genes per A.thaliana haploid genome. Sequence comparisons revealed a very conserved catalytic central domain flanked or not by four classes of extensions in the N- and/or C- extremities. The global amino acid composition of the extensions are tentatively correlated to specific functions such as targeting, protein interaction or RNA binding. The expression of the 32 AtRH genes has been recorded in different tissues. Separate patterns of expression and alternative polyadenylation sites have been shown. Based on the integration of all this information, we propose a classification of the AtRH proteins into subfamilies with associated functions. PMID- 9862991 TI - 5S rRNA gene deletions cause an unexpectedly high fitness loss in Escherichia coli. AB - In Escherichia coli, ribosomal RNAs (16S, 23S and 5S) are co-transcribed in a highly regulated manner from seven genomically dispersed operons. Previous studies on the cellular effects of altered levels of two of these rRNAs (16S and 23S) have been useful in better understanding the regulation of rRNA expression. Furthering these studies, we have investigated the effect of 5S rRNA deficiencies on cell fitness through the sequential deletion of 5S rRNA genes. Our findings indicate that the loss of 5S rDNA from multiple genes decreases cell fitness more rapidly than loss of a similar number of 16S and 23S rRNA genes. These results suggest that the cell's innate ability to up-regulate rRNA operons does not compensate for 5S rRNA deficiencies, as was previously shown for 16S and 23S rRNAs. A plasmid-borne 5S rRNA gene is able to compensate for the deleted 5S rRNA genes. PMID- 9862992 TI - Regulation by phosphorylation of the zinc finger protein KRC that binds the kappaB motif and V(D)J recombination signal sequences. AB - The DNA binding protein KRC (forkappaB binding andrecognitioncomponent of the V(D)J recombination signal sequence) belongs to a family of large zinc finger proteins that bind to the kappaB motif and contains two widely separated DNA binding structures. In addition to the kappaB motif, KRC fusion proteins bind to the signal sequences of V(D)J recombination to form highly ordered complexes. Here, we report that KRC may be regulated by post-translational modifications. Specific protein kinases present in the nucleus of pre-B cells phosphorylated a KRC fusion protein at tyrosine and serine residues. Such protein modifications increased DNA binding, thereby providing a mechanism by which KRC responds to signal transduction pathways. KRC is a substrate of epidermal growth factor receptor kinase and P34cdc2 kinase in vitro. Our results suggest that activation of the KRC family of transcription factors may provide a mechanism by which oncogenic tyrosine kinases regulate genes with kappaB-controlled gene regulatory elements. PMID- 9862993 TI - Immobilization of acrylamide-modified oligonucleotides by co-polymerization. AB - A flexible chemistry for solid phase attachment of oligonucleotides is described. Oligonucleotides bearing 5'-terminal acrylamide modifications efficiently co polymerize with acrylamide monomers to form thermally stable DNA-containing polyacrylamide co-polymers. Co-polymerization attachment is specific for the terminal acrylamide group. Stable probe-containing layers are easily fabricated on supports bearing exposed acrylic groups, including plastic microtiter plates and silanized glass. Attachment can be accomplished using standard polyacrylamide gel recipes and polymerization techniques. Supports having a high surface density of hybridizable oligonucleotide (approximately 200 fmol/mm2) can be produced. PMID- 9862994 TI - A novel assay for examining the molecular reactions at the eukaryotic replication fork: activities of replication protein A required during elongation. AB - Studies to elucidate the reactions that occur at the eukaryotic replication fork have been limited by the model systems available. We have established a method for isolating and characterizing Simian Virus 40 (SV40) replication complexes. SV40 rolling circle complexes are isolated using paramagnetic beads and then incubated under replication conditions to obtain continued elongation. In rolling circle replication, the normal mechanism for termination of SV40 replication does not occur and the elongation phase of replication is prolonged. Thus, using this assay system, elongation phase reactions can be examined in the absence of initiation or termination. We show that the protein requirements for elongation of SV40 rolling circles are equivalent to complete SV40 replication reactions. The DNA produced by SV40 rolling circles is double-stranded, unmethylated and with a much longer length than the template DNA. These properties are similar to those of physiological replication forks. We show that proteins associated with the isolated rolling circles, including SV40 T antigen, DNA polymerase alpha, replication protein A (RPA) and RF-C, are necessary for continued DNA synthesis. PCNA is also required but is not associated with the isolated complexes. We present evidence suggesting that synthesis of the leading and lagging strands are co-ordinated in SV40 rolling circle replication. We have used this system to show that both RPA-protein and RPA-DNA interactions are important for RPA's function in elongation. PMID- 9862995 TI - Folding of pyrimidine-enriched RNA fragments from the vicinity of the internal ribosomal entry site of hepatitis A virus. AB - Two RNA fragments from the region just upstream of the internal ribosome entry site of Hepatitis A virus (HAV) were studied, a 35mer (HAV-35), 5'U4C3U3C3U4C3U3C2UAU2C3U33(4), and a 23mer (HAV-23), 5(4)U4C3U3C3U4C3U33(4). Secondary structural predictions and nuclease digestion patterns obtained with genomic RNAs suggested that they link two stable Watson-Crick (WC) hairpins in the genomic RNA and do not form conventional WC secondary structure, but do fold to form a condensed, stacked 'domain'. To obtain more information, folding of HAV 23 and -35 RNA fragments was characterized using 1H nuclear magnetic resonance, in H2O as a function of pH and temperature, circular dichroism as a function of NaCl concentration, pH and temperature, and square-wave voltammetry as a function of pH. The results indicate that these oligo-nucleotides form intramolecular structures that contain transient U*U base pairs at pH 7 and moderate ionic strength (100 mM NaCl). This folded structure becomes destabilized and loses the U*U base pairs above and below neutral pH, especially at ionic strengths above 0.1. All of the cytidine protons exchange relatively rapidly with solvent protons (exchange lifetimes shorter than 1 ms), so the structure contains few if any C*CH+base pairs at neutral pH, but can apparently form them at pH values below 6. We present a series of possible models in which chain folding draws the strand termini closer together, possibly serving to pull the attached WC hairpin domains together and providing a functional advantage by nucleating reversible formation of a more viable RNA substrate. PMID- 9862996 TI - A detailed study of the substrate specificity of a chimeric restriction enzyme. AB - Recently, the crystal structure of the designed zinc finger protein, DeltaQNK, bound to a preferred DNA sequence was reported. We have converted DeltaQNK into a novel site-specific endonuclease by linking it to the Fok I cleavage domain (FN). The substrate specificity and DNA cleavage properties of the resulting chimeric restriction enzyme (DeltaQNK-FN) were investigated, and the binding affinities of DeltaQNK and DeltaQNK-FN for various DNA substrates were determined. Substrates that are bound by DeltaQNK with high affinity are the same as those that are cleaved efficiently by DeltaQNK-FN. Substrates bound by DeltaQNK with lower affinity are cleaved with very low efficiency or not at all by DeltaQNK-FN. The binding of DeltaQNK-FN to each substrate was approximately 2-fold weaker than that for DeltaQNK. Thus, the fusion of the Fok I cleavage domain to the zinc finger motif does not change the DNA sequence specificity of the zinc finger protein and does not change its binding affinity significantly. PMID- 9862997 TI - Catalytic and binding mutants of the junction-resolving enzyme endonuclease I of bacteriophage t7: role of acidic residues. AB - Endonuclease I is a 149 amino acid protein of bacteriophage T7 that is a Holliday junction-resolving enzyme, i.e. a four-way junction-selective nuclease. We have performed a systematic mutagenesis study of this protein, whereby all acidic amino acids have been individually replaced by other residues, mainly alanine. Out of 21 acidic residues, five (Glu20, Glu35, Glu65, Asp55 and Asp74) are essential. Replacement of these residues by other amino acids leads to a protein that is inactive in the cleavage of DNA junctions, but which nevertheless binds selectively to DNA junctions. The remaining 16 acidic residues can be replaced without loss of activity. The five critical amino acids are located within one section of the primary sequence. It is rather likely that their function is to bind one or more metal ions that coordinate the water molecule that brings about hydrolysis of the phosphodiester bond. We have also constructed a mutant of endonuclease I that lacks nine amino acids (six of which are arginine or lysine) at the C-terminus. Unlike the acidic point mutants, the C-terminal truncation is unable to bind to DNA junctions. It is therefore likely that the basic C-terminus is an important element in binding to the DNA junction. PMID- 9862998 TI - Enhancer of RNA polymerase III gene transcription. AB - A protein responsible for enhanced transcription by RNA polymerase III was identified in extracts from Xenopus oocytes. This protein, called EP3, interacts with a specific DNA sequence adjacent to the 3'-end of a Xenopus somatic 5S RNA gene and forms a distinct band shift complex with a unique DNase I footprint. Enhanced transcription was observed from both 5S RNA and tRNA reporter genes when EP3 binding sites were inserted at different locations and orientations. Removal of the EP3 protein from an oocyte extract abolished this enhanced transcription. In addition, EP3 was shown to stimulate transcription by increasing the rate of transcription complex assembly. EP3 directly discriminates between the somatic and oocyte 5S RNA gene families and may play a significant role in their differential expression during early Xenopus development. PMID- 9862999 TI - The integral divalent cation within the intermolecular purine*purine. pyrimidine structure: a variable determinant of the potential for and characteristics of the triple helical association. AB - In vitro assembly of an intermolecular purine*purine.pyrimidine triple helix requires the presence of a divalent cation. The relationships between cation coordination and triplex assembly were investigated, and we have obtained new evidence for at least three functionally distinct potential modes of divalent cation coordination. (i) The positive influence of the divalent cation on the affinity of the third strand for its specific target correlates with affinity of the cation for coordination to phosphate. (ii) Once assembled, the integrity of the triple helical structure remains dependent upon its divalent cation component. A mode of heterocyclic coordination/chelation is favorable to triplex formation by decreasing the relative tendency for efflux of integral cations from within the triple helical structure. (iii) There is also a detrimental mode of base coordination through which a divalent cation may actively antagonize triplex assembly, even in the presence of other supportive divalent cations. These results demonstrate the considerable impact of the cationic component, and suggest ways in which the triple helical association might be positively or negatively modulated. PMID- 9863000 TI - A method of one-step enzyme labelling of short oligonucleotide probes for filter hybridisation. AB - Here we describe a method of labelling short oligonucleotide probes with enzyme without purification or chemical modifications. Biotinylated oligonucleotides as short as 10 nt are coupled with streptavidin-conjugated enzyme, hybridised and detected with enzyme-triggered chemiluminescence. The detection of hybridisation signal is linear for two orders of magnitude of target dilution. It is shown to be comparable in sensitivity with standard procedures and with radioactive detection. The method is quick, simple and has potential for automation of large scale oligo-nucleotide hybridisation and multiplex sequencing. PMID- 9863001 TI - Rapid and efficient cloning of proviral flanking fragments by kanamycin resistance gene complementation. AB - We have developed a technique for the rapid cloning of unknown flanking regions of transgenic DNA. We complemented a truncated kanamycin resistance gene of a bacterial plasmid with a neomycin resistance gene fragment from a gene transfer vector. Optimized transformation conditions allowed us to directly select for kanamycin-resistant bacteria. We cloned numerous proviral flanking fragments from growth factor-independent cell mutants that were obtained after infection with a replication incompetent retroviral vector and identified integrations into the cyclin D2 and several unknown genomic sequences. We anticipate that our method could be adapted to various vector systems that are used to tag and identify genes and to map genomes. PMID- 9863002 TI - Tetracycline-inducible expression systems with reduced basal activity in mammalian cells. AB - We describe a modification of the tetracycline-inducible eukaryotic gene expression system with decreased basal levels of expression in HeLa cells. It employs the tetracycline-inducible transactivator and a tetracycline-regulated repressor fusion acting on the same promoter. To avoid heterodimerization or competition for the same DNA site, each was provided with different DNA recognition and/or protein dimerization specificities. We achieved active silencing in the uninduced state resulting in approximately 6-fold reduced levels of basal transcription and several hundred-fold activation of gene expression upon addition of tetracycline. PMID- 9863004 TI - Erucylphosphocholine, a novel antineoplastic ether lipid, blocks growth and induces apoptosis in brain tumor cell lines in vitro. AB - A potential benefit of alkylphosphocholines in brain tumor therapy was evaluated. The in vitro effects of the intravenously applicable erucylphosphocholine (ErPC) on proliferation, viability, morphology and cell cycle distribution of a rat glioma, four human astrocytoma/glioblastoma and a human medulloblastoma cell line were analyzed daily after continuous drug-exposure for up to six days. ErPC exerted strong cytostatic and direct cytotoxic effects on all cell lines tested at drug concentrations that are achieved in the rat brain after repeated intravenous injections of nontoxic drug doses. Concentrations of 70 microM (T98G, A172, 85HG66, 86HG39) and 110 microM (C6, D283 Med) led to complete cell death within 48-96 h. Particular characteristics of ErPC action are i) the accumulation of cells with a 4n DNA content corresponding to the G2/M-phase of the cell cycle, ii) the formation of two- and multinucleated cells and iii) the induction of apoptosis. PMID- 9863003 TI - p53-independent increase in E2F-1 expression enhances the cytotoxic effects of etoposide and of adriamycin. AB - The transcription factor E2F-1 drives cell cycle progression at the G1- to S phase boundary; however, overexpression of E2F-1 can induce apoptosis. We show here that E2F-1 protein levels increase in human medulloblastoma, glioma, lung, colon, and bladder cancer cell lines (n=7) following treatment with the DNA damaging agents adriamycin or etoposide. This induction of E2F-1 occurs independently of Rb or p53 status and involves new protein synthesis. Although E2F-1 protein levels increase following DNA damage, several genes transcriptionally targeted by E2F-1 are not similarly induced. Rather, induction of E2F-1 in the tumor cells correlates with their sensitivity to adriamycin or to etoposide. Correspondingly, fibroblasts from E2F-1 knockout mice are more resistant to DNA damage than cells from normal mice. Overexpression of E2F-1 protein in tumor cell lines infected with an adenovirus encoding wild-type E2F-1 leads to enhanced cytotoxicity following exposure to DNA damaging agents, which results from enhanced apoptosis. The results of this study implicate a role for E2F-1 in p53-independent cytotoxicity of chemotherapy and provide a pharmacological tool for increasing levels of the apoptosis-inducing E2F-1 protein. PMID- 9863005 TI - Constitutive and IFN-gamma regulated expression of IL-7 and IL-15 in human renal cell cancer. AB - Although not structurally related, the pleiotropic cytokines interleukin-7 (IL-7) and interleukin-15 (IL-15) share a variety of biological functions including stimulation and maintenance of cellular immune responses. Cytokines, such as IL-7 or IL-15, elaborated by cells in situ, e.g. cancer cells, may be involved in shaping the quality of anti-tumor directed immune responses. We have analysed the constitutive and IFN-gamma-inducible expression of IL-15 or IL-7 mRNA, protein expression, and protein secretion in human tumor cell lines of distinct origin. IL-15 mRNA expression was detected in renal cell carcinoma (RCC), small cell lung carcinoma (SCLC), glioblastoma, neuroblastoma, mesothelioma cells and in EBV transformed B-lymphocytes. IL-7-specific transcripts could be detected in colorectal cancer and in renal cell cancer cell lines. Immunohistochemical analysis demonstrated cytosolic IL-15 protein expression in renal cell cancer cells without apparent IL-15 protein secretion in vitro. Time kinetic analyses revealed that IFN-gamma mediated increase of IL-15 mRNA expression was transcriptionally regulated and dependent on de novo protein synthesis. However, enhanced IL-15 mRNA expression did not lead to effective protein secretion. In contrast, IL-7 mRNA expression in renal cell cancer or in colorectal cancer was associated with effective protein secretion which could be augmented by IFNgamma treatment. These data suggest that both IL-7 and IL-15 mRNA are expressed in renal cell cancer, but exclusively IL-7 may be elaborated by tumor cells in situ. IL-15 regulation appears to be tightly controlled both at the transciptional and post-transcriptional level. Appropriate stimuli leading to effective IL-15 secretion from tumor cells may aid in modulating cellular immune responses directed against cancer. PMID- 9863007 TI - Antitumor effect of vesnarinone on human hepatocellular carcinoma cell lines. AB - Vesnarinone, a quinolinone derivative, induces apoptosis and differentiation in some tumor cell lines. We examined the antitumor effect of vesnarinone on three human hepatocellular carcinoma (HCC) cell lines. Vesnarinone suppressed the proliferation of all three HCC cell lines, but did not induce apoptosis and differentiation. Vesnarinone has also been reported to inhibit both nucleoside and nucleobase transport. Concomitant administration of vesnarinone and a de novo nucleotide synthesis inhibitor potentiated the growth-inhibitory effect of vesnarinone on HCC cells. Vesnarinone may be useful as a new biochemical modulator of anticancer agents. PMID- 9863006 TI - Altered localization of E-cadherin and alpha-catenin in rat esophageal tumors. AB - An alteration in the localization of E-cadherin and its associated proteins has been observed in many epithelial neoplasms. No data exist, however, for the expression of these proteins in an animal model system for esophageal cancer or in cultured rat esophageal epithelial cell lines. The present study investigated the localization of E-cadherin and its associated protein, alpha-catenin, in rat esophageal epithelial cell lines of differing tumorigenic potential; in tumors induced after transplantation of these cell lines into syngeneic hosts; and, in esophageal tumors induced in rats by the carcinogen, N-nitrosomethylbenzylamine (NMBA). Immunofluorescent staining of the cultured cell lines revealed staining for both E-cadherin and alpha-catenin at cell-cell boundaries. Western blot analysis confirmed the membrane-bound localization of E-cadherin and alpha catenin in the cells. However, tumors induced by these cell lines in syngeneic rats showed reduction in the expression of both E-cadherin and a-catenin in the plasma membrane of invasive epithelial cells. Immunohistochemical analysis of NMBA-induced esophageal neoplasms in rats revealed E-cadherin and alpha-catenin to be abnormally expressed in poorly differentiated tumors when compared to well differentiated tumors. These results suggest that the microenvironment may have an important role in regulating the expression of these adhesion molecules in rat esophageal epithelial cells, and that alteration in the cellular localization of E-cadherin and alpha-catenin may be indicative of tumor progression in NMBA induced rat esophageal cancer. PMID- 9863008 TI - Expression of the thrombospondin 1 receptor CD36 is correlated with decreased stromal vascularisation in colon cancer. AB - Vascularisation is determined by the balance between various angiogenic and angio inhibitory factors in cancer stroma. CD36 is a cell adhesion receptor shown to interact with the ligand thrombospondin 1 (TSP1). It has not been determined whether the interaction of TSP1 and its receptor CD36 is correlated with vascularisation or clinical outcome of malignant tumours in vivo. We studied the correlations between expression of TSP1, CD36 and vascularisation and prognosis in 65 colon cancers. Various levels of TSP1 gene expression were observed in 27 of 65 colon cancers. CD36 expression was detected in 33 of 65 cancers, and was significantly correlated with decreased stromal vascularisation (P<0.001). The colon cancers expressing CD36 showed better prognosis (P<0.01). TSP1 expression levels did not affect CD36 expression. These results suggested that CD36 expression which decreased stromal vascularisation is correlated with better prognosis of colon cancer. PMID- 9863009 TI - Stable expression of activated Ki-Ras does not constitutively activate the mitogen-activated protein kinase pathway but attenuates epidermal growth factor receptor activation in human astrocytoma cells. AB - Mutation in the ras oncogene is one of the most commonly reported genetic aberrations in human cancer. Activated ras mutants are thought to play a major role in promoting the growth and malignancy of tumor cells. Ras protein plays a central role in transmitting mitogenic signals from cell surface-to-nucleus by activating signaling pathways in response to receptor activation. Ras protein by recruiting c-Raf-1 kinase to the plasma membrane activates the mitogen-activated protein (MAP) kinase pathway. Expression of activated ras mutants in rodent fibroblast has been reported to constitutively activate the MAP kinase pathway, suggesting that constitutive activation of this pathway contributes to Ras influence on proliferation and transformation. In this study, we investigated whether stable expression of an activated Ki-Ras oncogenic mutant (G12V) in human astrocytoma cells leads to constitutive activation of the MAP kinase pathway and how this may influence cellular proliferation and signaling by epidermal growth factor (EGF) receptor. We discovered that Ki-Ras stable expression does not lead to constitutive activation of the MAP kinase pathway, rather expression of Ki-Ras plays a role in attenuating the activation of this pathway in response to EGF stimulation. Furthermore, we provide evidence that stable Ki-Ras expression attenuates the ability of EGF receptor to activate the MAP kinase pathway by interfering with the receptor ability to autophosphorylate at tyrosine residues and not by down regulating receptor expression. PMID- 9863010 TI - Adenovirus-mediated over-expression of p21Waf-1 differentially affects the proliferation, telomerase activity, and expression of glial fibrillary acidic protein in human glioma cells. AB - Over-expression of p21Waf-1 was performed in 5 human glioma cell lines (U251MG, U87MG, T98G, NMC-G1, and KALS-1). p53 gene status was examined by PCR-SSCP and direct sequence method. The endogenous p21Waf-1 expression was evaluated by RT PCR. Adenovirus-mediated over-expression of p21Waf-1 inhibited the proliferation of human glioma cells expressing wild-type p53 (U87MG, NMC-G1, KALS-1) irrespective of endogenous p21Waf-1 expression. Telomerase activity was also reduced in these cells. In U87MG, GFAP expression was stimulated by the transfection with p21Waf-1, but was reduced in NMC-G1. Over-expression of p21Waf 1 thus affects the cellular behavior of human glioma cells in a cell-type specific manner. PMID- 9863011 TI - Serum levels of soluble intercellular adhesion molecule-1 and E-selectin in metastatic breast carcinoma: correlations with clinicopathological features and prognosis. AB - Cellular adhesion molecules have been demonstrated to play an important role in the progression and metastasis of malignancies. We determined the serum concentrations of soluble intercellular adhesion molecule-1 (sICAM-1) and E selectin (sE-selectin) by enzyme-linked immunosorbent assay in 49 Japanese patients with metastatic breast cancer. Significantly high concentrations of sICAM-1 and sE-selectin were found in the patients with liver and/or bone metastases (both P<0.05). The mean serum sICAM-1 levels were significantly higher in patients with two or more metastatic sites compared to those with one metastatic site (P=0.001). A significant correlation was found between serum sICAM-1 (P=0.0001) and sE-selectin (P<0.0001) and the interleukin (IL)-6 levels. The patients who did not respond to chemo/ endocrine therapy showed significantly higher sICAM-1 and sE-selectin levels compared with those who responded to therapy (P=0.0004, P=0.02, respectively). Moreover, high sICAM-1 levels predicted a significantly poorer overall survival in both univariate and multivariate analyses. Our results suggest that the shedding of sICAM-1 or sE-selectin may enhance the metastatic process by escaping from host immune surveillance. The serum sICAM-1 level may help to predict the patient response to chemo/endocrine therapy and may be of prognostic significance in metastatic breast cancer patients. PMID- 9863012 TI - Promoting activity of the herbicide dicamba (2-methoxy-3, 6-dichlorobenzoic acid) in two stage hepatocarcinogenesis. AB - Our goal was to examine whether dicamba, a widely-used broad leaf herbicide, has promoting activity in two-stage hepatocarcinogenesis. Female Sprague Dawley rats were given a single dose of diethylnitrosamine and then diets containing dicamba, or phenobarbital, or both for six months. The number and volume of placental glutathione-S-transferase-positive, glucose-6-phosphatase-negative or ATPase negative foci were quantified in the liver. Dicamba alone did not increase the number or volume of these altered hepatic foci. Dicamba did, however, show a significant effect on the number or volume of certain markers in animals also treated with phenobarbital. These data show that dicamba in combination with other promoters may have weak promoting activity in two-stage hepatocarcinogenesis in the rat. PMID- 9863013 TI - Apoptosis and Bbcl-2 expression in gastric carcinomas: correlation withclinicopathological variables, p53 expression, cell proliferation and prognosis. AB - We investigated apoptosis and Bcl-2 expression in 221 advanced gastric carcinomas in correlation with clinicopathological variables, p53 expression, cell proliferation and prognosis, using the in situ DNA nick end labeling method and immunohistochemistry. Apoptosis was associated with high immunoreactivity of proliferating cell nuclear antigen. Bcl-2 expression correlated with a low apoptotic index and less malignant behavior of tumors. Prognostically, Bcl-2 expression was associated with a better prognosis, whereas p53 expression was the most important prognostic risk factor. Thus, apoptosis in gastric carcinomas is associated with cell proliferation, and Bcl-2 expression may have a prognostic importance as well as p53 expression. PMID- 9863014 TI - TGFbeta-induced growth inhibition involves cell cycle inhibitor p21 and pRb independent from p15 expression. AB - It is generally assumed that TGFbeta induces cell cycle arrest through the cooperative action of cell cycle inhibitors p15, p27 and p21. Here, we found that several pancreatic carcinoma cell lines exert TGFbeta-induced negative growth control in spite of the loss of p15 and p16 expression. In these cell lines, TGFbeta-induced growth control correlates with the upregulation of the p21 protein and active pRb expression. Conversely, cells without p21 and/or pRb expression are resistant to TGFbeta -induced growth inhibition. Moreover, overexpression of p21 in the p21-deficient cell line Panc Tu1 leads to growth arrest. Thus, TGFbeta-induced growth control correlates with p21 expression and pRb status independent of p15 and/or p16 expression. PMID- 9863015 TI - DNA reactions, mutagenic action and stealth properties of polycyclic aromatic hydrocarbon carcinogens (review). AB - A brief summary of recent research, primarily from the authors' laboratory, on polycyclic aromatic hydrocarbon carcinogens with respect to their DNA adduct formation, the mutational properties of these adducts and the effects of hydrocarbon dihydrodiol epoxide metabolites on the passage of cells through the cell cycle is presented. The concept of stealth properties of potent carcinogens, i.e. their ability to damage DNA without inducing a G1 arrest, is discussed. Also, mutation studies with dihydrodiol epoxide metabolites, the sequence dependence of site-specific mutation, as well as the selectivity of hydrocarbon DNA adduct formation are summarized. PMID- 9863016 TI - Mutations in FL5.12 cells conferring resistance to apoptosis induced by interleukin-3 deprivation. AB - In an attempt to dissect the signal pathway in which Bax increases cellular responses to apoptotic stimuli and leads to the activation of the caspase cascade, we mutated FL5.12 Bax CL16 cells with a chemical mutagen. In this report we characterize two mutant clones, FL5.12 ms1 and m3. Both clones are resistant to IL-3 deprivation exhibiting no changes in mitochondrial membrane potential, annexin V and propidium iodide binding. FL5.12 ms1 is also resistant to staurosporine and anti-Fas antibody. In cell fusion experiments m3 behaves genetically dominant and ms1 is recessive. The results suggest that m3 has a mutation in a specific function upstream of Bax, while ms1 has a mutational block in the general pathway downstream of the 'Bcl-2 checkpoint'. PMID- 9863017 TI - Up-regulation of nuclear PP1alpha and PP1delta in hepatoma cells. AB - Activities of nuclear type 1 protein phosphatase (PP1) were significantly elevated in human HepG2 and rat AH13 hepatoma cells compared with primary cultured hepatocytes. We examined and compared the nuclear PP1 activities during the cell cycle between synchronized HepG2 cells and HGF-stimulated hepatocytes. Nuclear PP1 activity was significantly and more elevated at the G1/S transition in hepatoma cells compared with hepatocytes, although the amounts of PP1 isoforms remained constant. On the contrary, it was found that the basal levels of nuclear PP1 activity were significantly higher in hepatoma cells and that the amounts of PP1alpha and PP1delta were dramatically increased in the nuclear fraction of hepatoma cells. PMID- 9863018 TI - Mutation analysis of transforming growth factor beta type II receptor, Smad2, and Smad4 in hepatocellular carcinoma. AB - Mutations in the transforming growth factor beta type II receptor (TGFbetaRII), Smad2, and Smad4 genes have been detected in several human cancers. However, there are no reports of mutation analysis of the entire coding regions in these genes in hepatocellular carcinoma, and the roles of these genes in hepatocarcinogenesis remain unknown. We screened 30 hepatocellular carcinomas for mutations of these genes using polymerase chain reaction single-strand conformation polymorphism. We detected no mutations, but did find 3 cases of loss of heterozygosity of chromosome 17p13.1. These results suggest that mutations of the TGFbetaRII, Smad2, and Smad4 genes are rare, and that genetic instability is uncommon in human hepatocellular carcinoma. PMID- 9863020 TI - Enhancement of tumor uptakes by stabilized Fab homo-oligomers of a chimeric monoclonal antibody against carcinoembryonic antigen. AB - We investigated the effect of stabilized Fab oligomerization by disuccinimidyl suberate on tumor uptake in a pancreatic carcinoma xenograft model in nude mice. Recombinant mouse/human chimeric Fab of the anti-carcinoembryonic antigen (CEA) monoclonal antibody A10, which was previously shown to react specifically with gastrointestinal cancers was used in this study. Fab homo-oligomers (dimers and trimers) chemically linked with ethylene bonds (C-C oligomers) were produced by linkage of chimeric Fab. Oligomers with C-C bonds had similar immunoreactivity against human CEA to parental Fab monomer. In biodistribution studies in animals bearing pancreatic carcinoma xenografts, at 12 and 24 h after infusion, C-C oligomers showed significantly greater uptakes in tumors than Fab or F(ab')2 but lower than IgG. However, oligomers with C-C bonds maintained higher tumor to normal tissue specificity ratios than IgG 24 h post-infusion. In conclusion, tumor uptake was enhanced by Fab oligomerization with C-C bonds, compared to Fab or F(ab')2, perhaps due to the larger molecular size. It was also shown that C-C Fab oligomers could have a potency to deliver high-dose radionuclides with reduced radio-uptakes in normal tissues for the radioimmunotherapy of gastrointestinal carcinomas. PMID- 9863019 TI - Modulation of cytotoxic drug activity by mitotane and lonidamine in human adrenocortical carcinoma cells. AB - The ability of mitotane, a DDT derivative with adrenotoxic activity, and lonidamine, an energolytic derivative of indazole-carboxylic acid, to modulate the cytotoxic activity of doxorubicin, epidoxorubicin, cisplatin and VP16 was investigated in a human adrenocortical carcinoma cell line (SW13). A marked variability in cellular response to a 1-h treatment with the individual anticancer agents was observed. The concentrations able to inhibit SW13 cell proliferation by 50% (IC50) were 0.45 microg/ml and 0.4 microg/ml for doxorubicin and epidoxorubicin, respectively, thus indicating a relative sensitivity to anthracyclines. Conversely, the SW13 cell line displayed a marked resistance to cisplatin (IC50, 13.9 microg/ml) and VP16 (IC50, 15 microg/ml). When cells were exposed to anticancer drugs and mitotane simultaneously or in sequence, a positive modulation of anthracycline cytotoxic effects was observed. Although to a lesser extent, mitotane also increased cisplatin activity. Conversely, no potentiation was observed when mitotane was combined with VP16. Lonidamine slightly increased the cytotoxicity of epirubicin and cisplatin as individual agents. Moreover, a supra-additive effect of the three-drug (epidoxorubicin cisplatin-lonidamine) combination was observed. PMID- 9863021 TI - Monitoring of remission status by fluorescence in situ hybridisation in chronic myeloid leukaemia patients treated with interferon-alpha. AB - Interferon-alpha (IFN-alpha) can be considered as treatment of choice for patients with chronic myeloid leukaemia (CML) in chronic phase. With this treatment major cytogenetic responses can be achieved in 30% to 50% of patients. Regular monitoring of cytogenetic response is essential for the therapeutic management of these patients. As conventional cytogenetics is not always successful, especially under IFN-alpha treatment, molecular cytogenetic methods have been established for the examination of interphase nuclei for the presence of the BCR-ABL fusion gene, the molecular counterpart of the Philadelphia chromosome. To demonstrate the value of these new methods we have analysed interphase nuclei from sequentially cultured bone marrow cells from 14 CML patients who were treated with IFN-alpha and whose bone marrow was investigated regularly during therapy. Dual-colour FISH with a breakpoint spanning BCR-YAC and a flanking cosmid from the ABL region was applied. When compared with conventional cytogenetics the results achieved by FISH were favourable. The most evident advantage of FISH analysis is that in case of failure of conventional cytogenetics a reliable determination of the remission status can be done. Together with other recent studies our results illustrate the advantages and limitations of the interphase FISH method for monitoring CML patients. PMID- 9863022 TI - Microsatellite instabilities in gastric cancer patients with multiple primary cancers. AB - To answer whether microsatellite instability (MSI) can serve as a molecular marker for the risk-assessment of the development of multiple cancers, 26 tumors from 10 gastric cancer cases with multiple primary cancers were investigated. Six out of 10 cases revealed MSI in one or more cancer DNA. Significant statistical association was observed between MSI positive gastric cancer and cancer multiplicity (chi2 test, P<0.05). The complicated primary tumors in MSI-positive cases arose in colorectum, urogenital tract and ovary, which mimicked the tumor spectrum of hereditary non-polyposis colorectal cancer (HNPCC). On the other hand, most of the multiple cancers in MSI negative group were found synchronously and limited to the digestive organs. These observations suggest that MSI test on gastric cancer may be considered as a good marker for the assessment of multiple cancer development especially in the sites where tumors of HNPCC usually develop. PMID- 9863023 TI - The utility of a growth factor: r-HuEPO as a treatment for preoperative autologous blood donation in gynecological tumor surgery. AB - The use of r-HuEPO and sodium ferrous gluconate has been shown to be a safe and effective treatment which can be used by transfusional centers and surgeons to avoid allogeneic blood transfusions and to schedule short-term selective surgery. In this study the authors submitted 20 patients scheduled to undergo surgery for gynecological tumors to a program of pre-operative autologous blood donation. All the patients received both r-HuEPO and sodium ferrous gluconate in the pre- and post-donation period. r-HuEPO was administered subcutaneously in a dose of 200 IU/kg thrice weekly during the week before and after autologous blood donation (400 ml). Sodium ferrous gluconate was administered intravenously shortly before the first and fourth administration of 125 mg of r-HuEPO. Surgery was scheduled to be performed 10-15 days after the last r-HuEPo administration, i.e. within 15 20 days from blood donation. All the patients were tested for the following blood chemistry parameters: hematocrit, hemoglobin, sideremia and ferritin at treatment start, prior to donation, at treatment end, prior to autologous blood infusion and on the third and seventh day after surgery. No patient receiving r-HuEPO required allogeneic blood transfusion as both the hematocrit and hemoglobin values remained normal. r-HuEPO was observed to be a safe and effective treatment to be used in autologous blood donation programs in all patients scheduled to undergo surgery. It limits the decrease of hematocrit values following autologous blood donation thus enabling all the patients who for a variety of reasons to refuse allogeneic blood infusion to predeposit autologus blood shortly before the date scheduled for surgery. PMID- 9863024 TI - Quantitation by competitive PCR assay of vascular endothelial growth factor in non-small cell lung carcinomas. AB - The vascular endothelial growth factor (VEGF) is known to be one of the most important angiogenic factors under both physiological and pathological conditions. The VEGF overexpression by a wide spectrum of neoplastic diseases has suggested an important role of this cytokine in tumor-neovascularization. A method is described for quantification by reverse transcription-polymerase chain reaction (RT-PCR) of VEGF mRNA in non-small cell lung cancer tissues (NSCLC). The method entails addition to the sample of competitor DNA molecules that share the same primer recognition sites as the amplified target, but which can be easily distinguished by gel electrophoresis because of their different lengths (competitive PCR). We analyzed the VEGF mRNA expression level in 34 cases of lung tumor tissues compared to the respective adjacent normal tissues. In 4 out of 34 (11.7%) analyzed couples there was no VEGF mRNA expression, in 8 out of 34 (23.5%) only normal parenchymal tissue was positive for VEGF mRNA expression; in the remaining 22 cases (64. 7%) both normal and tumor tissues showed PCR products for VEGF. In 17 out of these 22 couples (77.2%) a higher number of VEGF mRNA molecules were present in tumor specimens than in the normal counterpart. According to these results, the competitive PCR-method seems to provide a useful tool for the quantitate VEGF expression in order to identify its role in the development of lung cancer. PMID- 9863025 TI - Elevated levels of urokinase-type plasminogen activator and its receptor during tumor growth in vivo. AB - Urokinase-type plasminogen activator (uPA) and its receptor (uPAR) are important in the regulation of tumor tissue progenesis, cell differentiation, tumor cell motility, and tumor cell invasiveness. We have recently reported that the levels of uPA and uPAR were higher in malignant astrocytomas than in low-grade gliomas. In the present study, we measured the levels of uPA and uPAR during the growth of glioblastomas in nude mice. Using fibrin zymography, densitometry, and an enzyme linked immunosorbent assay, we found that the enzyme activity and content of uPA were increased 4- to 10-fold during tumor formation. Using a receptor assay and an enzyme linked immunosorbent assay, we found the numbers and content of uPAR were increased 5- to 15-fold during tumor formation. In addition, immunohistochemical staining for uPA and uPAR revealed strong immunoreactivity in tumor cells with the staining more intense on day 28 than on day 14. These results suggest that the upregulation of uPA and uPAR plays a major role in the formation of gliomas. PMID- 9863026 TI - p53 protein overexpression is a prognostic indicator of poor survival in stage I endometrial carcinoma. AB - The prognostic value of p53 protein overexpression was investigated in a large series of early stage endometrial carcinomas with long follow-up (n=179, median follow-up 147 months). P53 overexpression was detected in 10 cases (5.6%). At the end of the study period, 30% (3/10) of patients with p53 protein overexpression had died of their disease compared to 6.5% (11/169) of those without overexpression. Multivariate analysis revealed that only myometrial invasion (RR 2. 1; 95% CI=1.2-3.6; P=0.001) and p53 overexpression (RR 9.5; 95% CI=2. 5-36.8; P=0.007) were independent predictors of survival. These results suggest that immunohistochemical evaluation of p53 protein overexpression provides strong prognostic information for the outcome of endometrial carcinoma patients with early stage disease. PMID- 9863027 TI - Somatic genetic alterations in human malignant mesothelioma (review). AB - A review of cytogenetic and molecular genetic findings in human malignant mesotheliomas (MMs) is presented. The complex profile of somatic genetic changes characteristic of MMs implicates a multistep process of tumorigenesis in this malignancy. In particular, the occurrence of multiple, recurrent cytogenetic deletions in MMs suggests that loss and/or inactivation of tumor suppressor genes (TSGs) are critical to the development and progression of such tumors. Karyotypic and comparative genomic hybridization analyses of MMs have demonstrated frequent deletions of specific regions within chromosome arms 1p, 3p, 6q, 9p, 15q and 22q, and subsequent loss of heterozygosity (LOH) studies have documented high frequencies of allelic loss from each of these chromosomal sites. Positional candidate gene approaches have identified TSGs within two of these regions, i.e., p16/CDKN2A at 9p21 and NF2 at 22q12, which are frequently altered in MMs. Homozygous deletions appear to be the major mechanism affecting p16/CDKN2A, whereas inactivating mutations coupled with allelic loss occur at the NF2 locus. High density LOH analyses have pinpointed minimal regions of deletion in 1p, 3p, 6q, and 15q and are expected to facilitate efforts to identify putative TSGs at these locations which contribute to the pathogenesis of MMs. PMID- 9863028 TI - Gene therapy: models to study the immunological way. AB - Gene engineering to enhance tumour immunogenicity and elicit curative responses against established tumours and tumour recurrences has become an attractive prospect. Gene engineering enables new genes to be selectively inserted into the genome of a tumour cell, or the construction of new fusion plasmids coding tumour antigens and immunomodulatory molecules. The rationale behind current research is to enhance the immune recognition of tumour antigens through their association with the molecules on which immune recognition depends. The immunotherapy data obtained in many experimental tumour systems provide a realistic assessment of the potential and limits of this technological approach. Experimental vaccination of rodents has been shown to induce a significant immune memory, even against poorly immunogenic tumours, that can prevent tumour growth and cure initial metastases, but is poorly effective against established tumours. Its use in tumour prevention is a fresh dawning perspective. PMID- 9863029 TI - Use of gene marking technologies in oncology. AB - Investigation of the mechanism of relapse in patients receiving stem cell rescue as therapy for malignant disease has been facilitated by gene marking studies. These studies have shown the marker gene to be present in malignant cells in the patient at the time of relapse, indicating that infused stem cells can contribute to disease recurrence. As normal progenitor cells are also marked and can be tracked in vivo, these studies have also helped us learn how haemopoietic stem cells respond to manipulation, for example with growth factors. Second generation studies with multiple, modified vectors are beginning to provide information about a wider variety of clinical and biological issues, including the efficacy of purging. Although marker studies have been useful for haematological malignancy and for neuroblastoma, they are hampered by the low efficiency of marking achieved by retroviral vectors. For many malignancies, marking efficiencies are insufficient for useful information to be obtained. This problem may be overcome by the introduction of vectors that, unlike retroviruses, can stably integrate in cells that are not in cycle at the time of vector exposure. Other improvements will focus on the marker genes themselves, using marker elements that are simpler to track and will not produce any modification of the cells' behaviour. Finally, marker studies have proved safe so far, but follow-up of the treated patients continues. PMID- 9863030 TI - The use of gene transfer in cancer immunotherapy. AB - A deeper understanding of the mechanisms underlying the regulation of immune responses together with the discovery of methods to identify tumour antigens have provided a strong foundation for the development of cancer immunotherapies. The recognition that multiple components of the immune system can effectuate tumour destruction has fostered the crafting of several strategies to augment anti tumour immunity. These approaches involve the stimulation of tumour antigen specific T lymphocyte and antibody responses, the augmentation of multiple components intrinsic to innate immune responses and the selective destruction of the tumour vasculature. A decisive factor in the crafting of these schemes has been the development of high efficiency gene transfer systems. These technologies render possible the genetic modification of a variety of cells playing critical roles in the evolution of anti-tumour immune responses; such modifications can dramatically enhance the levels of anti-tumour immunity. In this review, I will discuss the pre-clinical background underlying some of the current Phase I patient studies and highlight some of the intriguing early findings from these clinical investigations. PMID- 9863032 TI - Evidence-based medicine between explicit rationing, medical deontology and rights of patients. AB - Today a "just" health policy is balanced between the problem of the allocation of scarce resources and the priority setting of services, care and cures. Despite technologies and molecular medicine, with their tendency to reach absolute prediction of disease or absence of disease and to cure with predicted efficacy, a large portion of the public refuse the results of experimental procedures and prefer to place trust in so-called alternative medicine or in drugs which are not in the official guide-lines following the principles of evidence-based medicine according to DL Sackett. Juridical problems arise between the rights of free choice of cure and social dimension of Governmental care programs, which include the maximum of benefits (i.e. effective therapies) for a pre-fixed total budget. An explicit rationing only on budgetary bases without rationalisation of medical procedures reduces the rights to care of citizens-patients. Thus, an explicit rationing-rationalisation seems to be the only procedure compatible with the interest of patients in a social security system allocating "scarce" resources. PMID- 9863031 TI - Restoration of tumour suppressor gene expression for cancer. AB - Gene therapy has the potential to provide cancer treatments based on novel mechanisms of action with potentially low toxicities. This therapy may provide more effective control of loco-regional recurrence in diseases such as non-small cell lung cancer (NSCLC), as well as systemic control of micrometastases. Despite current limitations, retroviral and adenoviral vectors can in certain circumstances provide an effective means of delivering therapeutic genes to tumour cells. Although multiple genes are involved in the process of carcinogenesis, mutations of the p53 gene are the most frequent abnormality identified in human tumours. Pre-clinical studies both in vitro and in vivo have shown that restoration of p53 function can induce apoptosis in cancer cells. Phase I clinical trials now show that p53 gene replacement therapy is feasible and safe using both retroviral and adenoviral vectors, and that it induces tumour regression in patients with advanced NSCLC and recurrent head and neck cancer. Other pre-clinical studies indicate that gene therapy may have useful synergy with cytotoxic and radiation therapy. This paper describes the different gene therapy strategies under investigation and the pre-clinical data that provides a rationale for the gene replacement approach, reviews clinical trial data and presents novel ideas for improving current vectors and gene delivery to tumours. PMID- 9863033 TI - Natural rubber base matrix containing Calendula officinalis plant as a source of molluscicdal saponin. AB - Natural rubber (NR) has been used as a binding matrix for Calendula officinalis, where it is used as a vulcanizate. Various doses of the plant covering the range from 100 up to 200 parts per 100 of rubber (phr) were found suitable for processing conditions using conventional rubber mixing equipment. The release rate of saponin was extended to over 4 months. Incorporation of fillers, such as HAF and Hisil, in the compounded rubber decreased the release of saponin; especially in the first period of immersion. The amount of saponin released was affected by the environmental temperature and the type of fillers present in the formulations. The scanning electron micrographs illustrate the presence of a tortuous pass and pore structure that formed after soaking in water and leaching process occurred. PMID- 9863034 TI - Encapsulation of plasmid DNA in biodegradable poly(D, L-lactic-co-glycolic acid) microspheres as a novel approach for immunogene delivery. AB - A plasmid DNA encoding bacterial beta-galactosidase gene was encapsulated in poly(d,l-lactic-co-glycolic acid) (PLGA) microspheres. Plasmid DNA extracted from PLGA microspheres retained both structural and functional integrity as evidenced by its restriction endonuclease digestion pattern and its ability to transfect COS-1 cells in vitro. PLGA microspheres protected plasmid DNA from digestion by deoxyribonuclease I (DNase I) in vitro. The encapsulation efficiency of plasmid DNA and its release rate depended on the molecular mass of PLGA. Lastly, J-774A macrophages phagocytosed PLGA microspheres loaded with plasmid DNA. Co encapsulated monophosphoryl lipid A increased the rate of phagocytosis. These results suggest that biodegradable PLGA microspheres can deliver intact and functional plasmid DNA at controlled rates. Thus, PLGA microspheres may be used to jointly deliver genes and other biologically active molecules, e.g., immunomodulators, to antigen presenting cells. PMID- 9863035 TI - Evaluation of a positively charged submicron emulsion of piroxicam on the rabbit corneum healing process following alkali burn. AB - The effect of piroxicam in three different formulations was tested on rabbits for 28 days in an alkali burn model. The ulceration healing process was determined by evaluating the severity of the burn (scored from 0 to 5), and the re epithelization healing process was measured by the area of the defects. The results indicated that the piroxicam positively charged submicron emulsion was the most effective formulation in lowering the ulcerative cornea score while the piroxicam positively charged emulsion and the blank emulsion were more effective in promoting the re-epithelization healing process. The piroxicam solution elicited the slowest healing re-epithelization rate after 28 days and was unable to complete the entire healing process. The new positively charged submicron emulsion formulation of piroxicam had a pronounced effect on both the ulceration rate and epithelial defects in the management of corneal alkali-burning. PMID- 9863036 TI - Improved stability of highly fluorinated phospholipid-based vesicles in the presence of bile salts. AB - The stability of fluorinated phospholipid-based vesicles in terms of detergent induced release of encapsulated carboxyfluorescein has been evaluated. The fluorinated liposomes are substantially more resistant towards the lytic action of sodium taurocholate than conventional DSPC or even DSPC/CH 1/1 liposomes. Concerning structure/permeability relationships, the larger the fluorination degree of the membrane, the higher the resistance of the fluorinated liposomes to their destruction by the detergent. These results show that fluorinated liposomes have a promising potential as drug carrier and delivery systems for oral administration. PMID- 9863037 TI - Evaluation of mitoxantrone-loaded albumin microspheres following intraperitoneal administration to rats. AB - Mitoxantrone has demonstrated therapeutic efficacy in the regional treatment of intraperitoneal malignancies. However, severe local toxicity was dose limiting. Consequently, a new injectable sustained delivery formulation of mitoxantrone has been developed: the drug was incorporated (8.2 w/w%) in highly hydrophilic albumin microspheres. In vitro drug release profile was modified by matrix crosslinking extent. The extractable amount of residual crosslinking agent (glutaraldehyde) in the microspheres was lower than 6 ppm. Mitoxantrone concentration in peritoneal fluid and plasma was determined up to 72 h after intraperitoneal administration of 30, 60 and 120 mg mitoxantrone per m2 body surface area as solution and in the form of a dispersion containing mitoxantrone loaded microspheres to rats. Data analysis revealed sustained release of mitoxantrone from microspheres into peritoneal fluid in all dosage groups. The initial high drug levels in peritoneal fluid and plasma observed after application of mitoxantrone in the solution form were prevented by administration of the drug incorporated in microspheres. However, tumoricidal drug levels in peritoneal fluid were maintained over a comparable time span. In addition, preliminary toxicity data suggest a superior local tolerability of mitoxantrone loaded microspheres. The dose of intraperitoneally administered mitoxantrone might be increased from 30 to 60 mg per m2 body surface area using the slow release formulation. In conclusion, the described microsphere drug delivery system for mitoxantrone might overcome dose-limiting drug toxicity. PMID- 9863038 TI - In vivo drug release behavior in dogs from a new colon-targeted delivery system. AB - The colon-targeted delivery capsule (CTDC), a new capsule-type dosage form for colonic delivery of drugs, was investigated for the in vivo drug release behavior in dogs. A CTDC formulation with prednisolone as a model drug and theophylline as a marker substance for gastric emptying was prepared for this study. The enteric coated capsule (ECC) formulation with a similar composition was also prepared as the reference. Both formulations were administered to four beagle dogs, and the drug release behavior thereof was compared. Under fasted condition, ECC released prednisolone and theophylline at the same time within 1 h after the gastric emptying. On the other hand the CTDC released prednisolone at 3.2 h after the gastric emptying. Such release behavior of CTDC was approximately consistent with the results obtained from the in vitro dissolution study, suggesting that the pH sensing and timed-release functions imparted to the CTDC can work in the gastrointestinal tract of dogs as programmed. Under non-fasted condition, however, the gastric emptying of CTDC was found to be considerably delayed, up to about 14 h, and in this case the in vivo dissolution lag time of prednisolone at the small intestine was shortened to about 1.5 h. PMID- 9863039 TI - Asymmetric membrane capsules for osmotic drug delivery. I. Development of a manufacturing process. AB - This paper describes a novel non-disintegrating polymeric capsule and a manual and a semi-automatic process developed for its manufacture. The capsule wall was made by a phase inversion process in which the membrane structure was precipitated on stainless steel mold pins by dipping the mold pins into a coating solution containing a polymer-solvent-nonsolvent system followed by dipping into a quench solution. The resulting asymmetric membrane wall was composed of a thin dense region supported on a thicker porous region. The asymmetric membrane capsules can be filled with a blend of the active agent and excipients for use in osmotically modulated controlled drug delivery applications. PMID- 9863040 TI - Asymmetric membrane capsules for osmotic drug delivery II. In vitro and in vivo drug release performance. AB - In a previous paper, we described asymmetric membrane capsules and a phase inversion process for manufacturing them. In this paper, we describe the in vitro and in vivo drug release characteristics from these capsules. The capsule formulations were developed with model drugs to understand the variables that influenced drug release. Studies were also conducted to understand the drug release mechanism and it was shown that osmotic drug delivery was possible with asymmetric membrane capsules. PMID- 9863043 TI - Shuttle-vector system for Saccharomyces cerevisiae designed to produce C-terminal Myc-tagged fusion proteins. PMID- 9863041 TI - Influence of drug:hydroxypropylmethylcellulose ratio, drug and polymer particle size and compression force on the release of diclofenac sodium from HPMC tablets. AB - This study evaluates the relationship and influence of formulation and technological factors such as drug:hydroxypropylmethylcellulose (HPMC) ratio, particle size of the drug, particle size of HPMC and compression force, on drug release from matrices containing HPMC and diclofenac sodium as a model drug. The influence of these variables was assessed by multi-way analysis of variance. The results of the present study point out that the rate and mechanism of diclofenac sodium release from HPMC K15M matrices are mainly controlled by the drug:HPMC ratio. The drug and HPMC particle size also influence the drug release parameters, although to a lesser extent. Finally, the independence of the drug release from matrix tablets with respect to the compression force is reported. PMID- 9863044 TI - Characterization of tubulin isotype-specific antibodies by electrophoretic mobility shift assay. PMID- 9863045 TI - Stability of RNA molecules stored in GITC. PMID- 9863046 TI - Southern hybridization revisited; probe/target DNA interaction is affected by the choice of hybridization buffer. PMID- 9863047 TI - Electroporation of freshly plated Escherichia coli and Pseudomonas aeruginosa cells. PMID- 9863048 TI - PCR-based random mutagenesis method using spiked oligonucleotides to randomize selected parts of a gene without any wild-type background. PMID- 9863049 TI - PCR-based strategy for genotyping mice and ES cells harboring loxP sites. PMID- 9863050 TI - Adaptation of the fluorogenic 5'-nuclease chemistry to a PCR-based reverse transcriptase assay. PMID- 9863051 TI - Rapid isolation of HCV RNA from Catrimox-lysed whole blood using QIAamp spin columns. PMID- 9863052 TI - Colloidal silica-coated tissue culture dishes for primary cell cultures: growth of rabbit renal proximal tubule cells. AB - The use of colloidal silica as a substratum for primary cultures of differentiated cells has significant advantages over classic tissue culture polystyrene. In this report, the growth and the level of expression of differentiated function of primary rabbit renal proximal tubule (RPT) cell cultures on colloidal silica is examined, using hormonally defined serum-free medium. Primary RPT cells grew to confluence more rapidly on colloidal silica than on tissue culture polystyrene (TC+). Moreover, following three passages, the RPT cells increased in number threefold more than parallel cultures on TC+. The morphology of primary RPT cells on colloidal silica were found by means of transmission electron microscopy to possess a polarized morphology with a brush border, and differentiated markers were retained even after passaging, including the Na+/glucose cotransport system and Glut 7. PMID- 9863053 TI - Recombinant Bombyx mori nucleopolyhedrovirus harboring green fluorescent protein. AB - The gene encoding the green fluorescent protein (gfp) under the control of the highly expressed Autographa californica nucleopolyhedrovirus (AcMNPV)-polyhedrin promoter has been introduced into the polyhedrin (polh) locus of Bombyx mori nucleopolyhedrovirus (BmNPV) by homologous recombination. The insect host larvae and the cultured cells infected with this recombinant virus (vBmGFP) showed high levels of expression of gfp. The larval tissues permissive to virus multiplication could be readily visualized using the tagged recombinant virus, thus providing a direct approach to study the progress of virus infection or its control in the animal host. The highly expressed recombinant protein, GFP, could be easily solubilized from fat bodies. Thus, the caterpillar-based expression could serve as an economic alternative method for the large-scale production of recombinant proteins, even when they are nonsecretory in nature. Further, if the recombinant vBmGFP is used as a parent in generating other recombinants, conversion of the fluorescent plaques to colorless plaques serves as an easy means for screening recombinants. Such a method is especially helpful for BmNPV recombinant selections in the absence of the other simplified techniques as are available for the prototype baculovirus AcMNPV system. PMID- 9863054 TI - GFP:HIV-1 protease production and packaging with a T4 phage expression-packaging processing system. AB - A bacteriophage T4-derived protein expression, packaging and processing system was used to create recombinant phage that encode, produce and package a protein composed of human HIV-1 protease fused to green fluorescent protein (GFP). The fusion protein is targeted within the phage capsid by an N-terminal capsid targeting sequence (CTS), which is cleaved through proteolysis by the viral scaffold protease P21. The fusion protein is designated CTS [symbol see text] GFP:PR. The [symbol see text] symbol indicates the linkage peptide sequence leu(ile)-N-glu that is cleaved by the T4 head morphogenetic proteinase gp21 during head maturation. The fusion protein is fluorescent and has protease activity as detected by the appearance of the expected substrate cleavage product on a Western blot. CTS [symbol see text] GFP:PR packaging occurs at about 200 molecules per phage particle. The CTS [symbol see text] GFP:PR fusion protein, when protected within the phage capsid, has been maintained stably for over 16 months at 4 degrees C. Production and storage of fusion protein within the phage circumvents problems of toxicity and solubility encountered with E. coli expression systems. Because recombinant phage inhibit host proteolytic enzymes, foreign proteins are stabilized. This phage system packages and processes the fusion protein by means of the CTS. Proteins can be purified from the phage to give high yields of soluble, proteolytically processed protein. The T4 phage packaging system provides a novel means of identification, purification and long term storage of toxic proteins whose folding and DNA-directed activities can be studied readily in vivo. PMID- 9863055 TI - Site-directed mutagenesis of large plasmids. AB - A protocol combining recombination PCR and long-distance PCR is demonstrated to be highly accurate and rapid for site-directed mutagenesis of large (> 10 kb) plasmids. Application of this protocol to the generation of mutant rabies virus glycoproteins expressed by the baculovirus/insect cell system illustrates the usefulness of this approach in facilitating structure-function relationships in this important eukaryotic expression system. PMID- 9863056 TI - Subcycling-PCR for multiplex long-distance amplification of regions with high and low GC content: application to the inversion hotspot in the factor VIII gene. AB - Previously we described a PCR protocol for detecting the inversion in the factor VIII gene, which is a common cause of Hemophilia A. This PCR assay is challenging due to the size of the amplification (10-12 kb), the varying GC content (30%-80%) and the multiplex PCR products involved (four for carrier female). Efficient amplification of the four segments depends on three unusual modifications to standard long-distance PCR protocols: (i) very high concentrations of dimethyl sulfoxide, (ii) addition of deaza-dGTP and (iii) high concentration of Taq and Pwo DNA polymerases. One of the segments was amplified much more efficiently than the others under standard three-temperature cycling conditions (12 s at 94 degrees C, 30 s at 65 degrees C and 14 min at 68 degrees C). To facilitate the uniform amplification of the multiple regions, subcycling-PCR (S-PCR) was developed. In S-PCR, the combined annealing/elongation step is composed of subcycles of shuttling between a low and a high temperature, e.g., shuttling four times between 60 degrees and 65 degrees C. S-PCR produces consistent robust amplification of the various segments produced by wild-type, mutant and carrier individuals. S-PCR is a simple generalization of PCR, which generally may be advantageous in three contexts: (i) amplification of long segments in which the GC content varies within the segment, (ii) multiplex amplification of long segments and (iii) multiplex amplification of short segments in which the GC content varies among the segments. PMID- 9863057 TI - Chitobiase, a new reporter enzyme. AB - N,N'-diacetylchitobiase (chitobiase) from the marine organism Vibrio harveyi is a highly stable reporter enzyme for gene fusions. This enzyme hydrolyzes the disaccharide chitobiose to N-acetyl glucosamine. The advantages of the reporter gene encoding chitobiase (chb) are: (i) that chitobiase and N-acetyl-beta-D glucosaminidase activities are missing in E. coli strains, (ii) chitobiase can be monitored using blue/white colony indicator plates and (iii) convenient substrates for this enzyme are commercially available. The use of chitobiase as a reporter enzyme is generally applicable to the study of gene expression in those bacteria that do not contain N-acetyl-beta-D-glucosaminidases. We constructed plasmid vectors containing a multiple cloning site for producing in-frame fusions to chitobiase, the attP of lambda phase for movement into the bacterial chromosome for single-copy analysis, the gene encoding chloramphenicol acetyltransferase (cat), the pACYC184 origin of replication and the rrnBt1t2 terminator region upstream of the chb gene to prevent read-through from other promoters. In-frame fusions between the dnaA gene and chb were moved to the chromosome by site-specific recombination with the chromosomal attB site. These single-copy fusions were assayed for chitobiase to examine the effects of a deletion in the dnaA regulatory region. PMID- 9863058 TI - Degenerate oligonucleotide-primed PCR (DOP-PCR): evaluation of its reliability for screening of genetic alterations in neoplasia. PMID- 9863059 TI - Sensitive method for measuring telomere lengths by quantifying telomeric DNA content of whole cells. AB - Recently, a new method for measuring telomere lengths based on telomere DNA content was developed. The method, which is based on the ratio of telomere to centromere DNA content (TC ratio), is highly sensitive, allowing the analysis of small quantities of DNA. However, the method required the isolation of DNA, which can be difficult or impossible for small numbers of cells. Here, we suggest an improvement of this method that can directly estimate telomere lengths from whole cells. We optimized the method for whole cells and purified DNA and found that accurate TC ratios can be obtained from as little as 9 ng of DNA or 800 whole cells. There was no statistically significant difference between the ratios obtained with purified DNA or with whole cells, indicating that the isolation of DNA is not necessary for small samples. PMID- 9863060 TI - Telomerase assay using biotinylated-primer extension and magnetic separation of the products. AB - Human telomerase, a ribonucleoprotein enzyme is known to be associated with immortalized cancer cells but is absent in most normal tissues. Thus, telomerase appears to be an attractive new target for anticancer agents and an important diagnostic marker of human cancers. Here, we describe an improved telomerase assay method based on the Dynabead biomagnetic separation theory. In this method, 5'-biotinylated (TTAGGG)3 was used as a primer for the telomerase reaction. Telomerase reaction products were then immobilized on streptavidin-coated Dynabeads and washed intensively to eliminate excess [alpha-32P]dGTP. Using this method, without the amplification of telomerase reaction products by the PCR, we were able to quantitatively detect telomerase activity in human HeLa cell extracts equivalent to between 200-500 cells. This method is anticipated to be useful for the measurement of telomerase activity in various tumor cells, for assessing potential telomerase and for understanding the biochemical aspects of the telomerase reaction. PMID- 9863061 TI - Overexpression of human DNA topoisomerase II alpha by fusion to enhanced green fluorescent protein. AB - DNA topoisomerase (topo) II alpha is a major target for many anticancer agents. However, progress towards understanding how these agents interact with this enzyme in human cells and how resistance to these agents arises is greatly impeded by difficulties in expressing this gene. Here, we report on achieving a high level of expression of a full-length human topo II alpha gene in human cells. We started with the topo II alpha cDNA driven by a strong cytomegalovirus promoter and transiently transfected HeLa cells. Although topo II alpha mRNA was consistently detected in transfected cells, no exogenous topo II alpha protein was detected. By contrast, when the same cDNA was fused to an enhanced green fluorescent protein (EGFP), we detected a high level of expression at both mRNA and protein levels. The exogenous topo II alpha was localized to cell nuclei as expected, indicating that the fusion protein is properly folded. Furthermore, overexpression of the EGFP-topo II alpha fusion protein increased the sensitivity of the transfected cells to teniposide, suggesting that it functions as the endogenous counterpart. Thus, in addition to being used as a gene tag, the GFP fusion approach may be generally applicable for expressing genes, such as topo II alpha, that are difficult to express by conventional methods. PMID- 9863062 TI - Quantitative detection of t(14;18)-positive cells by real-time quantitative PCR using fluorogenic probes. AB - To detect t(14;18)-positive cells present in human lymphoma tissue, bone marrow aspirates and peripheral blood mononuclear cells (PBMNC), we have established an automated, real-time quantitative PCR using double-labeled fluorogenic probes. In relation to t(14;18)-positive genomic DNA or a cloned t(14;18)-DNA fragment, highly reproducible results can be obtained with initial copy numbers between 10 to 10(5). The detection of single copies has been verified by the stochastic multiple-tube approach. PBMNC cells obtained during clinical follow-up of patients with follicular lymphoma were analyzed by the one-step, real-time quantitative PCR and a two-step, semi-nested PCR combined with a limiting dilution assay. The quantitative results obtained by both assays correlate very well. Real-time quantitative PCR has several advantages: (i) it involves less critical pipetting steps, (ii) is less time-consuming and (iii) UTP, in combination with uracil-N-glycosylase, can be used to control carryover contamination. The higher specificity is due to optimized primer annealing conditions and MgCl2 concentration and the use of AmpliTaq Gold. The sensitivity is at least as high as by the two-step PCR. Real-time quantitative PCR will be very helpful in large epidemiological studies and in research for molecular staging and the detection of minimal residual tumor cells, including the analysis of blood stem-cell preparations to be used for transplantation after myelo ablative therapy. PMID- 9863063 TI - [Effects of green tea on growth inhibition and immune regulation of Lewis lung cancer in mice]. AB - C57/BL6J mice were inoculated with Lewis lung cancer cells as an experimental model to study the effects of green tea on cancer prevention, inhibition of tumor growth and immune regulation in mice with tumor. Results showed that weight of thymus in C57/BL6J mice and its index declined, proportion of positive CD4 subgroup of T lymphocyte and ratio of CD4+, to CD8+ reduced, baseline chemilumi nescence decreased in peripheral white blood cells, yeast zymosan stimulated chemiluminescence increased, and number of immunoglobulin M formation cells decreased. It indicated that green tea had obvious inhibition in Lewis lung cancer and protective effects, to various extent, on adverse changes of above indices. PMID- 9863064 TI - [Changes of coenzyme Q content in brain tissues of rats with fluorosis]. AB - Animal models with pathological damage caused by chronic fluorosis to a different extent were duplicated in Wistar rats by feeding them with 66.3 mg/L and 221 mg/L fluorine-containing water for three, five and seven months, respectively. Cholesterol, dolichol and coenzyme Q in animal brain tissues were determined by high performance liquid chromatography. Results showed that no significant difference of cholesterol and dolichol contents in brian tissues between rats with fluorosis and normal controls were detected. Coenzyme Q content of brain tissue in rats fed with fluorine-containing water decreased at early stage of fluorosis, but increased significantly at late stage. It is speculated that changes in content of coenzyme Q could correlate with changes in free radical levels induced by fluorine. PMID- 9863065 TI - [Cytotoxicity and genotoxicity of methyl tert-butyl ether and its metabolite to human leukemia cells]. AB - DNA damage of human leukemia (HL-60) cells caused by methyl tert-butyl ether (MTBE), a new gasoline additive, and its metabolites tert-butyl alcohol (TBA), a hydroxyisobutyric acid (HIBA) and formaldehyde was determined by single cell gel electrophoresis (SCGE), with release of lactate dehydrogenase as an indicator for evaluating its cytotoxicity. Results showed that MTBE, TBA and HUBA at levels of 1 to 30 mmol/L could cause DNA damage in a dose-dependent pattern. Formaldehyde at level of 5 mumol/L could cause DNA damage, but at a higher level could decrease DNA migration. It suggested that MTBE and its metabolites could have genotoxicity, however, with doses causing genotoxic effects, no cytotoxic effect by MTBE, TBA and HIBA was observed, but formaldehyde presented obvious cytotoxic effect. PMID- 9863066 TI - [Metabolism of aluminum in rats]. AB - In a study on metabolism of aluminium in rats, two doses of aluminium, 100 mg/kg and 40 mg/kg, were administered through gastric tube in SD rats to observe its storage, distribution in various tissues and 48-hour excretion in urine and feces. Results showed that storage and distribution of aluminium in various tissues were different in two doses during different time periods. Apparent absorption rates of aluminum were 19.07-19.45 and 3.50-7.72 percent and apparent retention rates were 16.89-20.67 and 3.76-13.48 percent 24 and 48 hours after its intake, respectively, in high dose group, and those in low dose group were 29.40 36.58 and 3.76-13.48 percent and 29.27-36.43 and 3.69-13.35 percent, respectively. In high dose group, contents of aluminium in liver 0.5 hours, in kidney one hour, in brain 48 hours after intake were higher than those in other time periods. The highest content of aluminum was in the liver, and the next in the brain and kidney, in low aluminium group, and that in brain of the rats 24 and 48 hours after intake in the high dose group were significantly higher than those in the low dose group (P < 0.01). It suggests that different doses of aluminium had certain affinity to the brain of rats. Cumulative excretion of aluminium in urine and feces accounted for 1.08-1.22 and 0.11-0.31, and 97.01 99.13 and 86.55-96.07 percent of the total intake, respectively, in the high and low dose groups. PMID- 9863067 TI - [Comparison of effects of two kinds of soluble algae polysaccharide on blood lipid, liver lipid, platelet aggregation and growth in rats]. AB - SD rats were fed with 1% cholesterol and 0.2% cholate for eight weeks. Sodium alginate and carrageenan, two kinds of soluble algae polysaccharide, were added to the fodder for the trial groups and none added for the control one. Results showed that 5% sodium alginate could lower lipid levels in the blood and liver and platelet aggregation (P < 0.05) in rats, but not affect their food intake and growth. Addition with 5% or 10% carrageenan had no obvious effects on lowering blood lipid and liver total cholesterol (P > 0.05), could not lower the ratio of liver weight to body weight, and had no stable (or sure) effect on lowering platelet aggregation. Ten percent of carrageenan could decrease their mean body weight by 12.17% in rats (P > 0.05), but a few rats suffered from slight diarrhea. PMID- 9863069 TI - [Studies on GBV-C infection in blood donors in four provinces of China]. AB - Plasma antibody against GBV-C (anti-GBV-C) and GBV-C RNA were detected with enzyme immunoassay (EIA) and reverse transcription nested polymerase chain reaction (RT-nPCR) in blood donors to study the prevalence of GBV-C infection in blood donors. Positivity of anti-GBV-C was 4.92 percent in 1,200 plasma donors (59/1,200), and that of GBV-C RNA was 64.41 percent in those with positive anti GBV-C (38/59). Positivity of anti-GBV-C was 2.86 percent in 350 whole blood donors (10/350) and that of GBV-C RNA was 60.00 percent in those with positive anti-GBV-C (6/10). Positive rate of anti-GBV-C in those who donated blood for more than 10 years was higher than in control group. It suggests that infection with GBV-C was more often in plasma and blood donors. There were blood donors with positive GBV-C RNA but normal ALT activities and those both positive in GBV C RNA and HCV RNA. It is urgent to detect the markers of GBV-C infection in blood donors as soon as possible. PMID- 9863068 TI - [A study on cell malignant transformation caused by dihydroxyglyoxime]. AB - BALB/c-3T3 cell transformation test was used to detect cell carcinogenicity caused by dihydroxyglyoxime (DHG) and determine its carcinogenicity in vitro. Results showed that DHG could induce transformation of BALB/c-3T3 cells and the transformed cells could agglutinate in the presence of Con A and grew in the soft agar. Transformed cells could induce the formation of fibrosarcoma if they were inoculated subcutaneously into the naked mice. It indicated sufficiently that the transformation in BALB/c-3T3 cell culture was malignant, and also predicted its potential carcinogenicity in human beings. PMID- 9863070 TI - [A study on viremia in patients with hepatitis E]. AB - Hepatitis E virus RNA (HEV RNA) was detected in 531 serial serum specimens collected from 62 hospitalized patients with hepatitis E by reverse transcription nested polymerase chain reaction (RT-nPCR) setting two pairs of primer from open reading frame 1 (ORF1) to understand the process of HEV infection more thoroughly. Results showed that 71 percent of the patients were positive for HEV RNA. Thirty-two of them with 288 serial serum specimens were followed up for the whole process, and 24 cases (75%) were positive for HEV RNA, positive rates declined with the course of the disease and serum HEV RNA lasted for 20.6 days in average. Serum HEV RNA was kept positive in 36 of the 44 patients (81.8%) at the time their serum alanine aminotransferase (ALT) and serum total bilirubin began to lower, and anti-HEV was detected in 39 of them (88.6%). Proportion of HEV viremia and length of it had no direct relationship with serum ALT and levels of total serum bilirubin and anti-HEV. PMID- 9863071 TI - [A prospective study on respiratory symptoms and functions in new employees exposed to cotton dust]. AB - Sixty new employees in a cotton textile mill were followed up for five years to study their occurrence of byssinosis and its natural history. Results showed that the workers complained more cough, expectoration and other respiratory irritation symptoms at their early exposure to cotton dust, and the frequency of chest tightness in them reached the peak one year after exposure and remained at higher level later. Stratified analysis showed that all the respiratory symptoms did not correlate with their smoking habits, specific body constitution, etc. Forced expiratory volume at one second (FEV1) after shift declined with length of exposure and appeared a tendency to exacerbate gradually, especially in workers with a positive skin test of cotton antigen. Smoking had no influence on it. It suggests that exposure to cotton dust and idiosyncracy of the workers play important roles in damage to lung function. PMID- 9863072 TI - [Effects of selenium supplement on acute lower respiratory tract infection caused by respiratory syncytial virus]. AB - An intervention study was conducted in 75 young children under one year hospitalized with pneumonia or bronchiolitis caused by respiratory syncytial virus (RSV) to evaluate therapeutic effectiveness of selenium (Se) supplement on acute respiratory lower tract infection caused by RSV with randomly controlled and double-masked method. Trial subjects were divided into two groups, one with 37 children in routine treatment and the other with 38 children in routine treatment plus Se supplement. The control group derived from 35 normal children during their physical check-up in the out-patient department. Sodium selenite was supplemented orally with 1 mg on the second day of hospitalization. Results showed that days needed for their relief of symptoms and signs were fewer in Se supplement group than that in controls and recovery in indicators of cell immune was better in the former than that in the latter. Levels of Se and glutathione peroxidase in plasma and white cells could be increased by Se supplement. It suggests that Se supplement can promote recovery from RSV infection. PMID- 9863073 TI - [A study on familial aggregation of hepatitis virus infection in Fujian province]. AB - In nine disease surveillance points of Fujian Province, positive serum antibody against hepatitis A virus (anti-HAV), hepatitis B surface antigen (HBsAg), antibody against hepatitis Cvirus (anti-HCV) and antibody against hepatitis E virus (anti-HEV) in people of 1,237 families were determined and their distribution was analyzed to study familial aggregation of hepatitis virus infection by fitting to binomial distribution. Results showed that chi 2 of goodness-of-fit to binomial distribution was 49.89, 217.91 and 27.39 for anti HAV, HBsAg and anti-HEV respectively, all with P < 0.001, indicating very obvious familial aggregation in HAV, HBV and HEV infection, with aggregation rates of 84.13%, 34.83% and 35.23% respectively. It suggests that familial aggregation is mainly caused by mother to infant transmission and horizontal transmission in families, and HEV infection can be transmitted through sex. PMID- 9863074 TI - [Progress in the study on health preserving and recovery by integrated traditional and Western medicine]. PMID- 9863076 TI - [Therapeutic effect of shuxuening combining neuroleptics for the treatment of chronic schizophrenia--a double blind study]. AB - OBJECTIVE: To assess the therapeutic effect of Shuxuening (SXN), the extractum of Ginkgo biloba761 (EGb761) in treating chronic schizophrenia. METHODS: A double blind, placebo-controlled multi-center research on the treatment of chronic schizophrenics with SXN was used. Five hundred and forty-five patients were randomly divided into either SXN group or the control group. Patients in the former group received SXN 120 mg three times daily. Patients in both groups received their maintenance neuroleptics throughout the 16-week research treatment. RESULTS: The patients' rating scores of brief psychiatric rating scale (BPRS) and Scale for the Assessment of Negative Symptoms reduced much greater in SXN group than those in the control group from the sixth week of treatment (P < 0.01). The effect of SXN for BPRS factors of retardation and thought disturbance was better than that of the control. SXN presented a better therapeutic effect for chronic schizophrenics than the control when rated with traditional global rating method as well, in which 44.98% marked improvement was obtained in SXN group compared to 20.98% in the control group. CONCLUSION: SXN combining neuroleptics, was an effective medicine for chronic schizophrenics. Moreover, it appeared few side-effects within the recommended dose range. PMID- 9863075 TI - [Clinical study of integrated traditional and Western medicine therapy on hepatolenticular degeneration]. AB - OBJECTIVE: To investigate the curative effect of integrated traditional and western medicine (TCM-WM) therapy on hepatolenticular degeneration (HLD). METHODS: Eighty patients with HLD were divided randomly into two groups (TCM-WM group and WM group), TCM-WM group (40 cases) were given orally dimercaptosuccinic acid (DMSA) and Gandou ([symbol: see text]) decoction for 1 month. The efficacy was compared with that of 40 cases treated with DMSA as the control. The changes of urinary trace and macro-elements before and after treatment were observed. RESULTS: The total effective rate of TCM-WM group was significantly higher than that of WM group (P < 0.05), especially more patients with hepatic type responded well to treatment with TCM-WM than with only chelating agent DMSA. Urinary trace and macro-elements were all obviously elevated in patients of two groups after treatment (P < 0.01). CONCLUSION: TCM-WM therapy appears extremely promising as new cupruretic method for treatment of HLD. It is particularly suitable for treating the patient with hepatic type. PMID- 9863077 TI - [Comparison on treatment of falciparum malaria with different courses of artesunate tablet]. AB - OBJECTIVE: To assess the efficacy of Artesunate on falciparum malaria. METHODS: A randomized controlled study on the treatment of 90 uncomplicated falciparum malaria patients was carried out with 400 mg of artesunate tablet as a total dose over 3 days, 600 mg over 5 days and 800 mg over 7 days. RESULTS: All patients were cured. Fever clearance time (FCT) and parasite clearance time(PCT) among the three groups were similar. Parasite recrudescence rate within 28 days was 39.3% (11/ 28) in 3 day group, 6.9% (2/29) in 5 day group and 3.4% (1/29) in 7 day group (comparing 5 day group with 3 day group, P < 0.005, comparing 7 day group with 3 day group, P < 0.005). CONCLUSION: It indicated that parasite recrudescence rate may be effectively decreased by prolonging treatment courses. PMID- 9863078 TI - [Effects of semen Cuscutae, rhizoma Curculiginis, radix Morindae officinalis on human spermatozoan's motility and membrane function in vitro]. AB - OBJECTIVE: To observe the effects of Semen Cuscutae (SC), Rhizoma Curculiginis (RC) and Radix Morindae Officinalis (RMO) on human sperm motility in vitro and cytomembrane function. METHODS: Compare the sperm motility of the above-mentioned medicinal plants with modified Tyrode's solution (MTS) and MTS added caffeine in vitro. Human spermatozoa were incubated with decoctions of medicinal plant at various concentrations in 37 degrees C for 30 minutes. The motility of sperm was evaluated by the sperm capillary penetrating test, sperm speed test and sperm activity index calculations. The spermatozoa membrane function was evaluated by spermatozoal hypo-osmotic swelling test together with eosin staining methods. RESULTS: The sperm motility improved markedly and sperm membrane function became more stabilized after incubation, the SC had best effect, RC took second place, RMO was relatively poor. CONCLUSIONS: SC decoction is an effective promoting preparation for sperm motility and the membrane function stabilized. Therefore, SC may be beneficial in treating male sterility and may raise success rate in artificial insemination. PMID- 9863079 TI - [Clinical study of combined acupuncture-drug anesthesia for anterior approach cervical discectomy]. AB - OBJECTIVE: To observe the effect of combined acupuncture-drug anesthesia for anterior approach cervical discectomy. METHODS: Fifty patients scheduled for anterior approach cervical discectomy were randomized into two groups, the control group (5% procaine combined with drug anesthesia, n = 25), and the experiment group (combined acupuncture-drug anesthesia, n = 25). In the experiment group, bilateral acupoints of Neiguan (P6) and Hegu (LI4) of the patients were stimulated for 30 minutes by Hans Acupoint Nerve Stimulator through skin electrode while a combination of dolantin 25 mg and droperidol 2.5 mg were administered intravenously during induction. RESULTS: There was no statistical difference of hemodynamic variation between the two groups (P < 0.05). The experiment group could also provide the same anesthesia effect as control group did, as well as it had less postoperative complications, more rapid recovery with less expense. CONCLUSIONS: The acupuncture-drug anesthesia for anterior approach cervical discestomy is feasible and it is worthwhile to be widely used in clinical practice. PMID- 9863080 TI - [Clinical and experimental studies on composite divitriol infusion in treating tinea manum]. AB - OBJECTIVE: To investigate the effect of Composite Divitriol Infusion (CDI) in treating tinea manum and studying its antimycotic action. METHODS: CDI was used to treat 139 patients with tinea manum. The experimental study in vivo was done, scanning electron microscopy (SEM) was used to observe the therapeutical effect of CDI. RESULTS: One hundred and four cases among the 139 patients were cured, 21 were markedly effective, 10 improved and 4 ineffective, the effective rate being 89.9%. Between CDI group and the control group, there was a very significant difference (chi 2 > 12.84, P < 0.005). The antimycotic action of CDI was studied in vitro. Its minimum inhibitory concentrations (MIC) of CDI were about 0.25% for Trichophyton rubrum, and 0.5% for Trichophyton gypseum and Microsporum gypseum. Trichophyton rubrum immersed with CDI show that the mycelia became roughened, deformed and macroconidum became smaller under SEM. CONCLUSION: CDI is valuable in treating tinea manum. PMID- 9863081 TI - [Effect of xuefuzhuyu pill on patients with carotid atherosclerosis by colour Doppler ultrasonography]. AB - OBJECTIVE: To observe the effect of Xuefuzhuyu Pill (XFZYP) on carotid atherosclerosis. METHODS: The patients were randomly divided into XFZYP group and aspirin control group. The status of the carotid plaque vessel wall and blood flow were observed by colored Doppler ultrasonography. RESULTS: (1) Plaque of 6 cases regressed, of 6 cases unchanged, of 1 case progressed after treatment of XFZYP. The area of plaques between baseline and after treatment decreased 0.11 +/ 0.05 cm and that of thickness reduced for 0.09 +/- 0.04 cm2 (P < 0.05) in XFZYP group. While plaque of 3 cases regressed, of 6 unchanged, of 2 cases progressed after treatment of aspirin. The area and thickness of plaque were reduced to 0.05 +/- 0.04 cm and 0.04 +/- 0.03 cm respectively. There were no statistic difference in aspirin group. (2) The intima-media thickness of common carotid artery decreased 0.13 +/- 0.03 mm in XFZYP group (P < 0.001 versus baseline), but there were no significant change after treatment of aspirin (P > 0.05). (3) There were no distinct change about peak systole, minimal diastole time average peak, pulsative index, resistive index in common carotid artery and internal carotid artery after treatment of XFZYP and aspirin. CONCLUSION: Colour Doppler ultrasonography of carotid artery provide a noninvasive, safe and valid opportunities for clinical trail. XFZYP can be useful for the treatment of carotid atherosclerosis. PMID- 9863082 TI - [Clinical and experimental study on treatment of chronic hepatitis B with yanggan aoping mixture]. AB - OBJECTIVE: To assess the efficacy of Yanggan Aoping Mixture (YGAPM) in treating hepatitis B. METHODS: Patients suffered from chronic hepatitis B were treated with YGAPM. Observe their short- and long-term efficacy and the change of serum hepatitis B virus marker. In experiment, the effect of YGAPM in treating rat's liver injury as well as HBV-infected tree shrew and duck HBV-infected ducks was observed. RESULTS: In 79 cases of chronic persistant hepatitis, the markedly effective rate was 60.76%, and follow-up studies on 40 cases, the further rasied to 70.00%. In 73 cases of chronic active hepatitis, the markedly effective rate was 60.27%, and further raised to 62.50% in 32 follow-up cases. In the treatment group, 85 (71.43%) of the 119 cases with HBeAg-positive turned to negative. Whereas in the control group, only 40 (44.94%) of 89 HBeAg-positive cases turned to negative, P < 0.01. Results of experimental study showed that negative conversion rate of tree shrew infected with HBV marker was raised, while infected duck blood with duck HBV DNA was inhibited. Those compared with the control group separately, the difference was remarkably significant. CONCLUSIONS: YGAPM is an effective drug in treating chronic hepatitis B, and it could effectively negative convert the HBV marker. PMID- 9863083 TI - [Study in the combining quantity of Con A-binding sites on membrane surface of artesunate-resistance strain of Plasmodium berghei]. AB - OBJECTIVE: To study the production mechanism of plasmodium berghei (PB) in resisting artesunate. METHODS: The combining quantity of Con A-binding sites on membrane surface of PB trophozoite labelled with fluorescein isothiocyanate (FITC) of Con A was determined by fluoromicrospectrophotometer. RESULTS: The combining quantity of PB artesunate-resistance strain (PBAR) was reduced significantly than that of PB normal strain (P < 0.01). CONCLUSIONS: The type of glucose in surface membrane of PBAR has changed and the changes of membrane fluidity could be one of the causes of the change in the combining quantity of Con A-binding sites on membrane of PBAR. PMID- 9863084 TI - [Experimental study on berberin raised insulin sensitivity in insulin resistance rat models]. AB - OBJECTIVE: To observe the effects of berberin on insulin sensitivity in high fat diet rats. METHODS: Before and after ingesting berberin, glucose insulin tolerance test was used to measure the insulin sensitivity, and determining fasting blood glucose, insulin, blood lipid and muscle triglyceride content. RESULTS: Compared with normal feeding rats, high fat feeding impaired insulin action (5.02 +/- 1.08 vs 8.72 +/- 0.91, P < 0.005), decreased liver glycogen level and raised fasting blood glucose, insulin and blood lipid levels. Berberin and metformin improved insulin resistance (6.31 +/- 0.95 and 6.24 +/- 0.65) and liver glycogen level in insulin resistance models, but had no effect on blood glucose, insulin, lipid levels and muscle triglyceride depots. CONCLUSION: It is confirmed that berberin could raise insulin sensitivity of high fat diet rats similar to metformin. PMID- 9863085 TI - [Effects of supplemented taohe chengqi decoction in treating insulin resistance in rats with non-insulin dependent diabetes mellitus]. AB - OBJECTIVE: To investigate the effect of Supplemented Taohe Chengqi Decoction (STHCQD) in treating non-insulin dependent diabetes mellitus (NIDDM). METHODS: The model of rats with NIDDM was formed with injection of streptozotocin and fed on high calorie diet to study the effects of STHCQD on the release of insulin mediator from liver cell membranes, the glucose oxidation in adipocytes as well as the insulin sensitivity. RESULTS: (1) Fasting serum glucose, serum insulin, intake of food and water were significantly decreased (P < 0.05-0.01) in STHCQD treated diabetic rats as compared with untreated diabetic rats, while the insulin sensitivity was significantly increased (P < 0.05). (2) The liver cell membranes from STHCQD-treated diabetic rats released the quantity of insulin receptor which inhibited adenylate cyclase activity, but this effect was blunted in untreated diabetic rats (P < 0.05). (3) A significantly increased glucose oxidation in adipocyte of STHCQD-treated diabetic rats was found as compared with those of untreated diabetic rats (P < 0.05). CONCLUSIONS: STHCQD therapy increased sensitivity and responsiveness of target cells to insulin, i.e., it might decrease insulin resistance at receptor sites and postreceptor sites in rats with NIDDM, but could not reverse the insulin resistance. PMID- 9863086 TI - [Effects of flos Carthami on beta-endorphin levels in brain regions of rats with cerebral ischemia]. PMID- 9863087 TI - [Survey and progress in the research of Chinese medicinal herbs intervention therapy in treating primary liver carcinoma]. PMID- 9863088 TI - [Effectiveness in the treatment of cerebral vascular disease]. PMID- 9863089 TI - [Progress in the study of integrated traditional Chinese medicine and Western medicine in the treatment of bronchial asthma]. PMID- 9863090 TI - [Clinical and experimental study on treating bronchial asthma with xingbei granule]. AB - OBJECTIVE: To explore the effect of the Xingbei Granule (XBG) in treating bronchial asthma and its mechanism. METHODS: Systematic clinical observation and animal experimental study on the XBG were carried out. RESULTS: The XBG not only could reduce the symptom of asthma, improve the ventilatory function effectively, but also has the effect of antagonizing the change of asthmatic inflammatory medium, alleviating the air-way's inflammatory reaction and lowering the air way's hyperreactivity. CONCLUSION: XBG do attain the dual goal of controlling the asthmatic symptoms and reducing the asthmatic attack. PMID- 9863091 TI - [Clinical study on jianfei pill in preventing and treating recurrence of infantile asthma]. AB - OBJECTIVE: Further explore on preventing and treating recurrence of infantile asthma with TCM method and its mechanism. METHODS: Using Jianfei Pill (JFP) which was prepared according to the therapeutic principle of strengthening the body resistance to consolidate the constitution and promoting blood circulation to remove obstruction in the channels in treating 56 patients suffering from asthma. Their pulmonary functions were observed. RESULTS: A total effective rate of 91.1% was shown in JFP group, which was higher than that of the control group (60%), P < 0.01. In comparison with the control group, the level of serum IgA, IgG and E rosette ascend significantly (P < 0.01). The wave amplitude of pulmonary impedance rheogram and right heart index (HI) also increased significantly (P < 0.01), while pulmonary arterial pressure (PPA) and pulmonary vascular resistance (PVR) dropped markedly (P < 0.01). CONCLUSION: JFP might work by means of enhancing the immune function and improving the pulmonary microcirculation to control chronic bronchial inflammation and reduce its reactivity to prevent the relapse of asthma. PMID- 9863092 TI - [Therapeutic effect of kangke injection on viral myocarditis and its anticoxsackie virus mechanism]. AB - OBJECTIVE: To assess the efficacy of Kangke Injection in treating viral myocarditis. METHODS: Kangke Injection is the effective ingredient extracted from Radix Sophora flavescens (RSF). Seventy-six cases of virus myocarditis suffering from the continuous positive Coxsackie B virus ribose nucleic acid-polymerase chain reaction (CBVRNA-PCR) in blood, their peak value in blood was determined by high performance liquid chromatography (HPLC) and compared with 50 cases treated by glucose-insulin-potassium chloride (GIK). RESULTS: The clearance rate of CBVRNA-PCR and RSF was dose-dependent. The effective rate of RSF on palpitation, chest distress, dispnea was 96.02%, and that of arrhythmia was 100%, all of them were better than those of control. After RSF therapy, the parameters of heart function of ejection fraction (EF), stroke volume (SV), cardiac output (CO), and cardiac index (CI) elevated significantly (P < 0.01), left ventricular mass (LVM) and left ventricular mass index (LVMI) were decreased statistically significantly, while after 5 month therapy, the anti-Coxsackie group B virus neutralizing antibodies of RSF group was returned to normal titer, natural killer (NK) cell activity elevated, P < 0.01. CONCLUSION: RSF was an effective substance for regressing the "Pathologic status" of viral myocarditis. PMID- 9863093 TI - [Clinical study of effect of yinji capsule on left ventricular systolic function of angina pectoris patients with blood stasis syndrome]. AB - OBJECTIVE: To explore the Yinji Capsule (YJC) in improving the left ventricular systolic function of angina pectoris patients with Blood Stasis Syndrome. METHODS: The systolic function of left ventricle (LV) in cardiac cycle of 28 angina pectoris patients with Blood Stasis Syndrome was examined with three dimensional echocardiograph (3-DE) before and after treatment with YJC. RESULTS: The total symptomatic effective rate was 85.7%. The changes of LV systolic function were those: left ventricle ejection fraction (LVEF) increased from 45.0 +/- 4.9% to 48.2 +/- 3.5% (P < 0.05); EF on early stage and late stage increased from 22.6 +/- 2.1%, 8.3 +/- 1.2% to 28.1 +/- 3.0% and 10.3 +/- 0.9% respectively (P < 0.01, P < 0.05), myocardial region with segment systole (SS) < 5% decreased significantly (P < 0.01). CONCLUSION: YJC could improve LV systolic function on early stage and late stage in cardiac cycles, and mainly improve the systolic function of the region with low SS of LV. PMID- 9863094 TI - [Clinical observation on aplastic anemia treated by Spatholobus suberectus Composita]. AB - OBJECTIVE: To investigate the pharmaceutical effect of Spatholobus Suberectus Composita (SEC) on bone marrow hematopoiesis and microenvironment of aplastic anemia. METHODS: One hundred and six cases of aplastic anemia patients were treated by SEC and the pathological change of bone marrow were observed before and after treatment for 24 months, and compared with 17 cases of normal volunteers as control. RESULTS: The total effective rate was 77.4%. The degree of marrow hyperplasia (15.7 +/- 10.2 vol% vs 39.2 +/- 13.9 vol%), the percentage of the granulocytes (31.4 +/- 12.4% vs 39.9 +/- 10.4%), the erythroblastes (19.8 +/- 15.9% vs 36.7 +/- 13.5%), megakaryocytes (1.4 +/- 2.3 cell/section vs 24.0 +/- 24.6 cells/section), and the microvessel number (40.0 +/- 23.4 ves/100 mm2 vs 201.0 +/- 141.0 ves/100 mm2) ascended significantly (P < 0.01 or P < 0.05). CONCLUSION: SEC could not only irritate the hematopoiesis, but also exert reconstructive action to marrow microenvironment. PMID- 9863095 TI - [Effects of ginkgo leave concentrated oral liquor in treating asthma]. AB - OBJECTIVE: To determine the effects of Ginkgo leave concentrated oral liquor (GLC) on airway inflammation. METHODS: Airway hyperreactivity and clinical symptoms and pulmonary functions of asthma patients were determined. RESULTS: In contrast to placebo group, GLC significantly reduced airway hyperreactivity (P < 0.05) and improved clinical symptoms (P < 0.05), pulmonary functions (P < 0.05) of the asthmatic patients. CONCLUSION: GLC is an effective drug of anti airway inflammation. PMID- 9863096 TI - [Clinical studies on liangxue piyan granule no. 2 in treating psoriasis]. AB - OBJECTIVE: To observe the effect of Liangxue Piyan granule No. 2 (LPG) on psoriasis. METHODS: Ninety-six cases of psoriasis patients were treated with LPG and a control group of 32 cases was set up, which was treated with Fufang Qingdai Capsule (FQC). RESULTS: The total effective rate of LPG was 84.7%, that of the control group was 71.9% (P > 0.05), there was no significant difference between two drugs in treatment of psoriasis. The recurrent rate of LPG group (3.6%) was lower than that of control (21.7%), P < 0.01. CONCLUSION: This granule provides a better drug for treatment of psoriasis. PMID- 9863097 TI - [Effect of theo-polyphenols on oxygen free radicals and hemorrheology in patients with essential hypertension]. AB - OBJECTIVE: To observe the effect of theo-polyphenols (TP) on oxygen free radicals and hemorrheology in patients with essential hypertension. METHODS: Hemorheology as well as oxygen free radical of 38 patients were determined before and after TP treatment. RESULTS: TP markedly increased RBC-SOD activity, reduced the level of serum MDA, the whole blood viscosity, plasma viscosity and the level of plasma fibrinogen. The efficacy of TP was higher than that of aspirin. CONCLUSION: The TP had remarkable effects of anti-oxygen free radicals, and could improve hemorheology. PMID- 9863098 TI - [A preliminary study on effect of electro-acupuncture by the cortical infrared thermography]. AB - OBJECTIVE: To explore the possibility of using cortical infrared thermography (CIT) and temperature in studying the principle of acupuncture and moxibustion. METHODS: The changes of the cerebral cortical temperature during electro acupuncture (EA) by CIT in twenty cats were observed. By opening skull before experiments the cortex of animals was revealed. The distribution of the cortical temperature of the awakend animals was studied by AGA Infrared Thermovision, the thermograms were analysed by computer. RESULTS: (1) The thermograms of the profile of the cortex could be displayed clearly, the temperature was higher and lower in ante- and post-cortex respectively than that in other regions. (2) The cortical temperature declined slightly during 30 minutes in cats without EA. (3) After EA, the temperature of cortex generally increased 0.3 to 1.7 degrees centigrade, at 5 min after EA, the temperature increment was the highest, at 10 min after EA, the temperature increment was higher than before EA but lower than 5 min after EA, the cortical temperature recovered 10 min after stopping stimulating. (4) The temperature increment was more obvious in the regions of Ant. Ectosylvian, Ant. Suprasylvian and Ant. Sylvian than that in other regions. No matter where the site of EA was, every group had its region of higher temperature increment. The regions of the temperature increment in the cortex were mainly in the somatesthesia regions. There were similar characteristics between the temperature response in the cortex showing by CIT after EA and the activity of cortical nerve cells after stimulation. CONCLUSION: The display of the temperature response could be taken as the indication of the nervous activity. CIT could be used to conduct the studies of the acupuncture and brain. PMID- 9863099 TI - [Effects of coumarins from cnidium monnieri on the function of pituitary adrenocortical axis in kidney yang deficiency rats]. AB - OBJECTIVE: To observe the effect of Cnidium monnieri in Kidney Yang Deficiency rats. METHODS: Rats were injected intraperitoneally with hydrocortisone acetate to replicate the animal model of Kidney Yang Deficiency. Osthol, total-coumarins (TCR) from the fruit of Cnidium monnieri and positive control drug (PCD) were administered orally to model rats. The plasma levels of corticosterone (B), adrenocorticotropic hormone (ACTH), renin (PRA), angiotensin-II (AT-II) and aldosterone (ALD) were determined by radioimmunoassay. RESULTS: Compared with those in normal rats, the levels of B and ACTH in the plasma of model rats lowered significantly (P < 0.01), the levels of B and ACTH of treatment groups were higher than those of model group, while the levels of PRA, AT-II and ALD had no significant difference (P > 0.05). CONCLUSION: The osthol, TCR have the effects in protecting and strengthening the function of pituitary-adrenocortex axis. PMID- 9863100 TI - [Effect of qiangji jianli capsule on metabolism of acetylcholine receptor]. AB - OBJECTIVE: To explore the action mechanism of Qiangji Jianli Capsule (QJJLC) treating myasthenia gravis. METHODS: Internal receptor permeation and degradation test were performed. RESULTS: Volume of receptor permeation in the group with QJJLC at 96 hours was 902.50 +/- 37.48 cpm/mg, while that of control group 738.45 +/- 35.41 cpm/mg. Half-life of receptor degradation in treatment group was 52 hours and that of control 38 hours. Their difference was very significant (P < 0.001). CONCLUSION: QJJLC could accelerate permeation of N-aectylcholine receptor to surface of diaphragm cell membrane and decelerate receptor degradation. It could also increase accumulative amounts of functional receptor in unit time and keep dynamic equilibrium of receptor metabolism in pathologic condition. PMID- 9863101 TI - [Inhibitive effect of umbellatus polyporus polysaccharide on cachexic manifestation induced by toxohormone-L in rats]. AB - OBJECTIVE: To investigate the effect of treatment of cachexia with Umbellatus Polyporus Polysaccharide (UPP). METHODS: UPP was used to treat cachexia induced by toxohormone-L. RESULTS: Toxohormone-L from ascites fluid with primary hepatoma could induce lipolysis in vitro significantly, inhibit the intake behavior, decrease the level of blood zinc, increase that of blood copper (P < 0.01), upp could inhibit these effects. CONCLUSION: Toxohormone-L was closely related with neoplastic cachexia and UPP could inhibit this effect induced by toxohormone-L significantly. PMID- 9863102 TI - [Present status of researches on pulmonary hypertension]. PMID- 9863103 TI - [Present status and prospects of researches on type A behavior]. PMID- 9863104 TI - [Exploration of research on important fields of basic theory of traditional Chinese medicine]. PMID- 9863105 TI - [Clinical study on ligustrazine in treating myocardial ischemia and reperfusion injury]. AB - OBJECTIVE: To explore the protective effect of Ligustrazine in treating myocardial ischemia, and reperfusion injury. METHODS: The activities of serum superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and lactic dehydrogenase (LDH) and the amount of malondialdehyde (MDA) as well as the effects of Ligustrazine (LGT) on them were determined in 16 patients with cardiopulmonary bypass, who were, scheduled for elective cardiac surgery, were randomly divided into control group and LGT group. Ligustrazine was given by intravenous drip within 2-3 minute with a definite speed before occlusion and immediately after release respectively. Their venous blood samples were collected to measure the serum levels of SOD, GSH-Px, LDH and MDA by biochemical methods before the occlusion of aorta, at 30 minutes of occlusion and at 30 minutes after release respectively. RESULTS: There were significantly and very significantly differences between the values of control group and LGT group. CONCLUSION: LGT could effectively protect the myocardium from ischemia and reperfusion injury. PMID- 9863107 TI - [Clinical observation on 206 cases with lower limb deep venous thrombosis treated by integrated traditional Chinese and Western medicine]. AB - OBJECTIVE: To study the pathogenetic mechanism of lower limb deep venous thrombosis (DVT). METHODS: Thrombosis test in vitro of 206 cases with lower limb DVT was measured. All of them were treated by integrated traditional Chinese and Western medicine (TCM-WM). RESULTS: The indexes of thrombosis test in vitro of patients were much higher than those of normal subjects (P < 0.001). The relevent indexes were different in types distribution according to differentiation of symptoms and signs (P < 0.01-0.001) and between before and after treatment (P < 0.001). It showed that hypercoagulability of blood occurred even if after the occurence of DVT. Changes of indexes might be objective parameters for Syndrome Differentiation in TCM. The indexes became lower when state of illness was improving. CONCLUSION: The therapy of TCM-WM has good efficacy for DVT. PMID- 9863106 TI - [Clinical and experimental study on jinshui liujun decoction in treating chronic bronchitis]. AB - OBJECTIVE: To assess the effect of Jinshui Liujun decoction (JSL) in treating chronic bronchitis. METHODS: The effect of JSL on expelling the phlegm and strengthening the body resistance were observed. RESULTS: JSL was proved to be efficacious in improving immune function of the body and pulmonary ventilatory function (P < 0.01). JSL could not only increase the secretion of rats' trachea mucosa and dilute sputum, but also increase the motion speed of the pigeons' trachea cilia, thus facilitate the removal of sputum. Compared with the normal saline control group, both animal experiments displayed significant statistical difference (P < 0.01), but without significant difference when compared the JSL group with the positive drug Juhong Tanke Ye control group. CONCLUSION: JSL has the effect of resolving phlegm as well as improving human immunological and pulmonary ventilatory functions. PMID- 9863108 TI - [A report on 8 seronegative converted HIV/AIDS patients with traditional Chinese medicine]. AB - OBJECTIVE: For the first time, serum anti-HIV antibody negative conversion was being reported. METHODS: Eight confirmed HIV/AIDS patients (AC1, ARC 6, AIDS 1) were treated with TCM recipes (802, 806, 809, 810, Shengmaiyin, ZY-1), recheck the serum antibody and immunological function and intranuclear HIV-DNA was investigated with PCR amplifying assay and were long-term followed-up. RESULTS: After medication for 87-463 days, seronegative conversion occurred, PCR assay revealed that 5 cases were PCR(+), 2 of them(-), 1 turned seropositive again in the early stage. Observed continuously for 11-49 months, the "serum negative and intranuclear positive" state maintained. These patients belonged to immunosilent HIV-infection. The immunological function of all seronegative converted patients were good. CONCLUSIONS: AIDS is a reversible disease. Using medicinal herbs to enhance the immune function will facilitate the appearance of seronegative conversion, which has not been reported before. If it could be further confirmed, its mechanism elucidated, this may greatly strengthen the confidence of the patients. PMID- 9863109 TI - [Clinical and experimental study on rapid bladder ultrasound developer of Chinese medicinal herbs]. AB - OBJECTIVE: To seek for a rapid type B ultrasound developer of Chinese medicinal herbs, so that the bladder and pelvic cavity developed clearly and pelvic cavity diseases could be diagnosed rapidly. METHODS: One hundred and twenty-two patients were observed clinically and animal experiments were performed. The rapid bladder ultrasonography developer (RBUD-1, a preparation of Chinese herbal medicine) alone was used in Group 1, composite prescription of Western and Chinese medicine was used in group 2. The control groups were using lasix or mineral water. RESULTS: Rapid diuresis and the decrease of the bladder capacity needed for development could be realized by Chinese medical herbs preparation, the difference between Group 1 and control group in developing time and bladder capacity were very significant. Results of animal experiments, which were referred to clinical grouping, showed the diuretic intensity of RBUD-1 within one hour was significantly higher than that in the other groups. Toxicological study showed the RBUD-1 was a non-toxic preparation. CONCLUSION: RBUD-1 could effectively develop bladder and pelvic cavity, it would help to diagnose in time, on the other hand, it would also contribute for the combination imaging of Chinese and Western medicine. PMID- 9863110 TI - [Clinical and experimental study on improving cellular immunological function of uterine myoma patients by xiaoliu tablet]. AB - OBJECTIVE: To explore the effect of Xiaoliu Tablet (XLT) in treating uterine myoma (UM) and the relationship between effect and the cellular immunological function of the UM patients. METHODS: Seventy-one UM patients by treatment with XLT was observed. The activities of natural killer (NK) cells of 50 patients and 30 normal women were determined. RESULTS: There were good effect in reducing the volume of myoma and improving the menses by XLT, especially in patients whose myoma were singly and small. The activity of NK cells of these patients (24.61 +/ 6.04%, n = 50) was significantly different from that of normal women (45.42 +/- 9.05%, n = 30), P < 0.01. There was a notable rise of NK cells of the patients (37.54 +/- 5.01%, n = 39), after the treatment of six months by XLT (P < 0.01). CONCLUSION: Improving the immunological function might be one of the mechanisms of the effect of XLT. PMID- 9863111 TI - [Clinical observation on effect of xiaoyu zhixue tablet on 104 patients with idiopathic multifocal bleeding and platelet aggregation defect]. AB - OBJECTIVE: To explore the effect and pharmacological mechanism of Xiaoyu Zhixue Tablet (XZT). METHODS: The XZT mainly composed of Chinese medicinal herbs to replenishing Qi and removing stasis was used to treat patients with idiopathic multifocal bleeding and platelet aggregation defect. Patients were divided into 2 groups. The TCM group (104 cases) treated with XZT, and the control group (84 cases) treated with vitamins C, K and P, adrenosin and (or) artificial menstruation, for at least 4 months. RESULTS: The hemostatic effective rate was 92.3% in TCM group and 27.4% in control group (P < 0.005), the recovery rate of platelet aggregation after 6 months therapy was 69.2% and only 1.8% respectively. The difference between two groups was very significant (P < 0.005). CONCLUSION: XZT could regulate the physiological functions of blood-vessel and platelet to realize the hemostatic action and platelet aggregation. PMID- 9863112 TI - [Clinical study of sudden deafness treated with method of integrated traditional Chinese and Western medicine]. AB - OBJECTIVE: To explore the mechanisms of sudden deafness treated with integrated traditional Chinese and Western medicine (TCM-WM). METHODS: One hundred and forty six patients suffering from sudden deafness were divided into two groups, 64 cases of treated group treated with the method of integrated TCM-WM and the other 82 cases treated with Western medicine alone for control. RESULTS: The total effective rate of the treated group was 92.19%, that of the control group was 75.61%. The difference between two groups was significant, P < 0.01. CONCLUSION: The integrated method is very effective in treating sudden deafness. Its mechanism might be: (1) Decrease the blood viscosity and increase the red blood cell's tenacity, so that the blood stream is passing through unimpeded. (2) Decrease the plasma TXA2 level and increase the PGI2 level, keep the blood supply for inner ear. (3) Increase the activity of SOD and clear away the free radicals so as to reduce the damage of inner ear. PMID- 9863113 TI - [Effect of acupoint irradiation with Q-wave millimeter microwave on peripheral white blood cells in post-operational treatment with chemotherapy in stomach and colorectal cancer patients]. AB - OBJECTIVE: To explore the biological effect of Q-wave millimeter microwave (QWMM). METHODS: The QWMM was used to irradiate the acupoints Xuehai (Sp10) and Geshu (B17) in treating post-operational and chemotherapy treated stomach cancer and colorectal cancer patients. The effect of irradiation on chemotherapy affected peripheral white blood cells was observed. 62 cases (stomach cancer 42, colorectal cancer 20) in total were divided into two groups: group A, 21 cases (stomach cancer 15, colorectal cancer 6) the irradiation began 10 days after operation, and on the 16th day the chemotherapy combined with irradiation started. Group B had 41 cases (stomach cancer 27, colorectal cancer 14), in which the irradiation began immediately after the occurrence of chemotherapy induced peripheral WBC reduction, which persisted for at least 12 days. RESULTS: The effective rate for the group A and B was 85.7% (18/21) and 73.2% (30/41) respectively. The total effective rate of the two groups was 77.4% (48/62). The effective rate of group A was significantly higher than that of group B, P < 0.01. CONCLUSION: GWMM irradiation at acupoints could promote the hematopoietic function of bone marrow, and the irradiation performed 1 week before chemotherapy yielded even better protection on bone marrow. PMID- 9863114 TI - [Experimental study on effect of bushen shengjing decoction on kidney yang and testicular dysfunction in rats]. AB - OBJECTIVE: To explore the therapeutical mechanism of Bushen Shengjing Decoction (BSSJD) in treating infertility. METHODS: Fed Wistar male rats with adenine 30 mg/100 g.d continuously for 30 days. The rats manifested the symptom which was similar to the human Kidney Yang Deficiency, the genitality dropped, the function of sperm-producing in the testis was disturbed, the contents of luteinizing hormone (LH), testosterone (T) and compound F (F) in blood were all lower than rats of normal group, P < 0.01. The model rats took the BSSJD 3 ml/100 g.d (each ml contains crude drugs 2 g) continuously for 10 days, interval for three days, then medicated again and total medication of BSSJD was 30 days. RESULTS: The symptom of Kidney Yang Deficiency in rats was improved obviously, the genitality, sperm quality and function of sperm producing in testis were all enhanced obviously, so did the hormone level of LH, T, F in blood. Comparing to untreated control group, the difference was significant (P < 0.05). CONCLUSION: BSSJD has the function of reinforcing Yang and replenishing essence and regulate the endocrinological function. PMID- 9863115 TI - [Observation of therapeutic effect by combined administration of Salvia miltiorrhiza, ligustrazine and Panax notoginseng on late hemorrhagic shock of rabbits]. AB - OBJECTIVE: To explore the therapeutic mechanism of Salvia miltiorrhize, ligustrazine and Panax notoginseng in treating late hemorrhagic shock in rabbit. METHODS: Rabbit hemorrhagic shock models (MPA 5.3 kPa) were set up according to Wiggers' method and administrated Salvia miltiorrhiza, ligustrazine, Panax notoginseng. The values of blood RBC superoxide dismutase (SOD) and blood lactate (BL), plasma malondialdehyde (MDA) and magnesium (Mg++) were continuously monitored before shock, 120 minutes after shock, 60 and 120 minutes after hydraulic dilatation. RESULTS: (1) In 120 minutes after shock, the level of SOD decreased and the concentrations of MDA, BL, Mg++ were markedly increased, which indicated that the cell membrane damage caused by oxygen free radicals in rabbit hemorrhagic shock. (2) Salvia miltiorrhiza, Ligustrazine or Panax notoginseng could alleviate lipidperoxidation injury to tissue. Compared with the single drug administration groups, the effects of oxygen free radicals scavangers by combined administration with half dose of 2 drugs were better than the single drug with full dose alone and the side effects such as depression of blood pressure and heart rates would be alleviated. CONCLUSION: Combined administration of Salvia miltiorrhiza, ligustrazine and Panax notoginseng would half the dosage, the blood pressure depression and heart rate reduction alleviated and better result obtained. PMID- 9863116 TI - [Experimental study on effect of shenyi xieli mixture on nature killer cell activity and lymphocyte reactivity to IL-2 in tumor mice]. AB - OBJECTIVE: To explore therapeutic mechanism of Shengyi Xieli Mixture (SYXLM) on malignant tumor. METHODS: Chemotherapy, radiotherapy and different doses of SYXLM were used to treat tumor mice [correction of rats]. RESULTS: SYXLM could enhance the nature killer (NK) cell activities of the tumor mice [correction of rats], and this effect was dose dependent (control group 52.70 +/- 6.01, SYXLM No. 1 group 59.01 +/- 10.00, SYXLM No. 2 group 78.33 +/- 10.90, P < 0.01). The NK cell activities of the tumor mice [correction of rats] were inhibited in the chemotherapy group alone, but they were enhanced in the group of chemotherapy combined with SYXLM (P < 0.05), the results were the same in the radiotherapy group alone and the group of radiotherapy combined with SYXLM (P < 0.05). The response of lymphocyte of the tumor mice [correction of rats] to IL-2 was enhanced significantly in SYXLM group compared with the control group. This response was enhanced insignificantly in the group in which radiotherapy combined with the SYXLM. CONCLUSION: The SYXLM could enhance the function of immune system of the tumor mice [correction of rats] and diminish or reduce the side effect of radiotherapy, chemotherapy to the immune system of the tumor mice [correction of rats]. PMID- 9863117 TI - [Effect of electroacupuncturing heart meridian on arterial blood pressure with myocardial ischemia in rabbits]. AB - OBJECTIVE: To explore the relationship between Heart Meridian (HM) and cardiac function. METHODS: Forty-eight rabbits with acute myocardial ischemia resulting from intravenous pituitrin injection was conducted. Electroacupunture(EA) on the HM and the Lung meridian, 3 points for each, was applied and their change of arterial pressure was observed. RESULTS: The distinct difference existed in the improvement of arterial pressure on the HM and Lung Meridian with EA. The regulatory action of EA in HM on the cardiac function was significantly better than that of EA in Lung Meridian. CONCLUSION: HM was closely related to the cardiac function. PMID- 9863118 TI - [Status quo of experimental and clinical studies in retarding kidney damage of chronic nephropathy by ligustrazine]. PMID- 9863120 TI - [Traditional Chinese medicine in comprehensive therapy of neoplasm]. PMID- 9863119 TI - [Progress in experimental and clinical study on extracorporal screening drugs of anti-hepatitis B virus with 2.2.15 cell line]. PMID- 9863122 TI - [Therapeutic effect and its mechanism exploration on mainly using traditional Chinese medicine of replenishing qi and nourishing yin in treating Graves disease]. AB - OBJECTIVE: To explore the therapeutic effect and its mechanism mainly using traditional Chinese medicine (TCM) of replenishing Qi and nourishing Yin (RQNY) with a small dosage of Tapazol for treatment of Graves disease (GD). METHODS: The changes of thyroid function and the activity of sodium pump of human erythrocyte in the patients with Graves disease were observed and compared before and after treatment between the treated group (42 cases) by combining treatment mainly using TCM of RQNY and a small amount of Tapazol, and a control group (42 cases) by Tapazol alone. RESULTS: After treatment for half a year, one and two years, the serum levels of T3, T4 in above two groups were markedly decreased than those of before treatment, the therapeutical effect of treated group was superior to that of control group. The activity of sodium pump in human erythrocyte in the GD patients was obviously higher than that of normal group and that of before treatment. After treatment for one and two years mainly by TCM or Western medicine, the erythrocyte sodium pump activity was obviously lower than that of before treatment and that of normal group. The decrease of erythrocyte sodium pump activity in group of combination therapy was markedly lower than that in group of Western medicine. CONCLUSION: Combination therapy was much more effective on the functional remission of thyroid and energy metabolism in GD patients than that of using Tapazol therapy only. PMID- 9863121 TI - [Clinical observation on 112 cases with non-Hodgkin's lymphoma treated by Chinese herbs combined with chemotherapy]. AB - OBJECTIVE: To seek for the effective therapeutical method in treating non Hodgkin's lymphoma (NHL). METHODS: One hundred and sixty seven patients with non Hodgkin's lymphoma were randomly divided into two groups, the treatment group, which consisted of 112 cases using Chinese herbs combined with chemotherapy and 55 cases of control group were treated by chemotherapy only. RESULTS: The effective rate (CR + PR) in the combined group was 91.96% and survival rates of 1 , 3- and 5-year were 85.7%, 54.5% and 29.5% respectively, and median survival time was 554 days. In control group the effective rate was 72.73% and 1-, 3- and 5-year survival rates were 76.4%, 38.2% and 18.2% respectively, and the median survival time was 465 days. The difference of effective rates or 3-year survival rates between two groups was significant (P < 0.05). In the combined group the activity of NK cell, OKT3, OKT4 and ratio of OKT4/OKT8 were obviously raised after treatment (P < 0.01). And the level of platelet adhesion rate and the blood viscosity markedly decreased (P < 0.01), but in the control group the values of these indexes did not distinctly change. CONCLUSION: Chinese herbs could enhance the immunologic function and improve the viscosity of blood of the patients with NHL. The side effect in the combination therapy group was less and milder than that in the chemotherapy group. These showed that Chinese herbs combined with chemotherapy was a safe and effective method for treating NHL and deserve to be recommended. PMID- 9863123 TI - [Comparative study of applying tiaoxue yisui recipe and SSL regimen in treating infantile chronic aplastic anemia and analysis of its therapeutical mechanism]. AB - OBJECTIVE: To explore the effective method in treating infantile chronic aplastic anemia (ICAA) by using traditional Chinese medicine (TCM). METHODS: Seventy-eight cases of ICAA were observed, 48 of them were treated with Tiaoxue Yisui recipe (treated group), 30 cases were treated with SSL regimen (control group). RESULTS: The remission rate and total effective rate in treated group were 52.08% and 81.25% respectively, they were higher than those in control group (P < 0.05). After one year treatment the ratio of hemopoietic and non-hemopoietic cells in treated group was higher than that in control group (P < 0.05). The number of megakaryocyte in treated group was more than those in control group (P < 0.05). CONCLUSION: Tiaoxue Yisui recipe could improve the quality of the patient's life. The therapeutical mechanism of the Tiaoxue Yisui recipe might promote the proliferation of hemopoietic stem cells and regulate the immune function. PMID- 9863124 TI - [Clinical study on effect of bushen shengxue paste in treating renal anemia patients]. AB - OBJECTIVE: To evaluate the therapeutical mechanism of Bushen Shengxue Paste (BSSXP) on anemia. METHODS: Chronic renal failure induced anemia patients were treated with BSSXP, clinical manifestation, anemia and renal function as indicators were observed in patients. The erythropoietin (EPO) and inhibition of colony-forming unit-erythrocyte (CFU-E) in patients' serum were determined by CFU E in vitro. RESULTS: The patients' symptoms, renal function and anemia were improved after administration with BSSXP 1-2 course. EPO in serum was slightly increased. The inhibition of CFU-E in patient's serum was significantly decreased. CONCLUSIONS: BSSXP could improve the anemia degree, its mechanism might be through clearing the inhibitor of CFU-E in serum. PMID- 9863125 TI - [Effect of Salvia miltiorrhizae Composita on erythrocyte membrane phospholipid in patients with coronary heart disease]. AB - OBJECTIVE: To investigate the function of phospholipid metabolism in pathogenesis of coronary heart disease (CHD) and the effect of Salvia miltiorrhizae Composita (SMC) on it. METHODS: The levels of erythrocyte membrane phospholipid, blood lipid peroxide (LPO), superoxide dismutase (SOD) and glutathione peroxidase (GSH Px) were determined in the patients with CHD by Thinlayer chromatography and fluorimetry during the treatment of SMC. RESULTS: Sphingomyelins (SM), phosphatidyl serine (PS), phosphatidyl ethanolamine (PE) and lysophosphatide (LL) of erythrocyte membrane phospholipid were increased in patients with CHD. Phosphatidyl cholines (PC) was lower than control group. Among them, the ratios of SM/PC and PS/PC were significantly higher. In the patients with CHD, LPO was significantly increased and ratios of SOD/LPO and GSH-Px/LPO significantly decreased. After the treatment with SMC, LL, SM, PS and PE of erythrocyte phospholipid and the ratios of SM/PC and PS/PC and LPO were decreased. PC and SOD/LPO were increased. CONCLUSIONS: The abnormality of erythrocyte membrane phospholipid participate the occurrence and development of CHD. SMC might improve the abnormality of erythrocyte phospholipid and lipid peroxide and possess the effect of regulating phospholipid metabolism. PMID- 9863126 TI - [Effect of puerarin on plasma endothelin, renin activity and angiotensin II in patients with acute myocardial infarction]. AB - OBJECTIVE: To study the changes of endothelin (ET), renin activity (RA) and angiotensin II (AT-II) before and after puerarin treatment in patients with acute myocardial infarction (AMI). METHODS: Forty-three patients with AMI were divided into two groups, and were given puerarin and glucose-insulin-kalium (GIK) treatment respectively. Plasma ET, RA and AT-II were measured by radioimmunoassay (RIA) before and after treatment in different phases. RESULTS: It showed that plasma ET and RA, AT-II levels in AMI were higher than those in control group (P < 0.01). ET level was conversely correlated with RA and AT-II (P < 0.01). After treatment with puerarin, plasma levels of ET, RA and AT-II were recovered to normal in 3 days, but these data recovered to nearly normal until 7-14 days in group with GIK treatment. CONCLUSION: Puerarin might play an important role in regulating the imbalance of ET, RA and AT-II of patients with AMI. PMID- 9863127 TI - [Effect of xinmaitong capsule on damage of lipid peroxidation in coronary heart disease patients with myocardial ischemia]. AB - OBJECTIVE: To assess the effect of Xinmaitong Capsule (XMTC) in treating coronary heart disease(CHD). METHODS: The superoxide dismutase(SOD) and malonyldialdehyde(MDA) levels, ST of electrocardiogram and clinical symptoms of 25 CHD patients with myocardial ischemia were observed before and after treatment with XMTC and 15 patients with Composite Danshen Tablet (CDT) were studied as controls. RESULTS: The SOD level was significantly lower than that of normal (P < 0.001), and MDA significantly increased (P < 0.001). After treatment with XMTC, the SOD levels and ST of ECG were significantly increased (P < 0.001). In the control group, the SOD levels and ST of ECG were unchanged (P > 0.05), MDA and scores of clinical symptoms were decreased (P < 0.05, P < 0.001), but the levels of them were markedly higher than those in the XMTC group (P < 0.01). CONCLUSION: Lipid peroxidation injury is closely related with the myocardial ischemia and Syndrome Differentiation of TCM, XMTC with function of supplementing Qi and activating blood circulation could resist lipid peroxidation injury. PMID- 9863128 TI - [Improvement of sini decoction on hemorheology following percutaneous transluminal coronary angioplasty]. AB - OBJECTIVE: To study the hemorheological effects of Sini decoction on patients following percutaneous transluminal coronary angioplasty (PTCA). METHODS: Forty six patients were randomly divided into Sini decoction and control groups. The hemorheologic variables were determined before and after Sini decoction treatment. RESULTS: No hemorheologic changes were observed in the patients (n = 23) only with PTCA, but the patients (n = 23) with Sini decoction were found to be significantly decreased in whole blood viscosity and red cell aggregation and dredging the blood of microcirculation as post-PTCA compared to pre-PTCA. CONCLUSION: Sini decoction could improve the patient's hemorheology. PMID- 9863129 TI - [Effect of glucosidorum Tripterygii tororum on interleukin-6 and interleukin-8 on human peripheral mononuclear cells of healthy subjects]. AB - OBJECTIVE: To explore the effect of glucosidorum Tripterygii tororum (GTT). METHODS: Using different techniques of cell culture, the function of GTT in comparing with that of hydrocortison (HC) on interleukin (IL)-6 and -8 production by peripheral mononuclear cells of healthy Subjects was investigated. RESULTS: (1) Both of GTT and HC inhibited IL-6, IL-8 production, the effect of GTT being stronger when the dosage was higher than 1 microgram/ml; (2) In the isolated cells culture, the effect of GTT was similar to that of HC, but in the whole blood culture, the effect of GTT was much weaker than that of HC. CONCLUSIONS: (1) GTT inhibited interleukin production in a dose-dependent manner; (2) The sensitivity of interleukin to drugs was different in different culture conditions; (3) The anti-inflammatory effect of GTT was related to the interleukin inhibition, but the mechanism might be different from that of HC. PMID- 9863130 TI - [Study of different pattern of immuno-modulatory mechanism by tonifying kidney and invigorating spleen]. AB - OBJECTIVE: To observe the acting pattern on the immune system by tonifying the Kidney (TK). METHODS: Three composite recipes were used on the 7- and 14-day corticosterone rat model to study the effect of recipes on the different pattern of immuno-modulatory and neuroendocrinological systems. RESULTS: The immune system of Invigorate the Spleen(IS) group was well protected in either 7-day or 14-day experiment, without any effect of IS recipe acting on the neuroendocrine system, it suggested that IS recipe might have direct action on immune system, while TK group with no effect on neuroendocrine-immune (NEI) system in 7-day experiment, but showed the remarkable protective effect of the whole NEI system until 14-day experiment. CONCLUSION: The results indicated that TK recipe is acting on the neuro endocime system first, and then influence the immune system. It means that the action was through down pathway of NEI network. PMID- 9863131 TI - [Effects of composite blood-activating decoction on adhesive function of bone marrow stroma cells in radiation injured mice]. AB - OBJECTIVE: To evaluate the effect of commonly used "blood activating and stasis removing" Chinese herbal drugs on adhesive function of bone marrow stroma cells in acute radiation injured mice. METHODS: After irradiation by 8 Gy 60Co gamma ray, each mouse was intraperitoneally injected immediately with 0.2 ml 100% composite blood-activating decoction (CBAD) twice a day for 6 days. On the 7th day, the femura were taken and the bone karyocyte (BMC) suspension was made. According to long term bone marrow culture procedure, on the 12th day, the fibroblastic colony forming units were counted, and on the 28th day, the adhesion of culture stroma cells to normal murine BMC was observed. RESULTS: The adhesion of culture stroma cells to normal murine BMC was 67.8 +/- 17.2% in normal group, while in CBAD group, it was 55.8 +/- 10.5%. The difference was insignificant (P > 0.05). In the control group, the value was 47.7 +/- 13.6%, which was significantly lower than that in normal group (P < 0.02). CONCLUSION: The commonly used "blood-activating and stasis-removing" Chinese herbal drugs can enhance the adhesive function of bone marrow stroma cells in acute radiation injured mice. PMID- 9863132 TI - [Effects and mechanism of emodin and sandostatin on pancreatic ischemia in acute haemorrhagic necrotizing pancreatitis]. AB - OBJECTIVE: To investigate pancreatic ischemia and abnormal metabolism of eicosanoids in acute haemorrhagic-necrotizing pancreatits (AHNP) and the effects of emodin or sandostatin on them. METHODS: Rats with AHNP were triggered with sodium taurocholate; the pancreatic blood flow (PBF) was detected with computerized tissue blood flowmeter, and plasma prostaglandin E2 (PGE2), 6-keto prostaglandin F1 alpha (6-keto-PGF1 alpha) and thromboxane (TXB2) were determined with radioimmunoassay. RESULTS: There was a significant decrease of PBF in the early stage of AHNP. Compared with that in the untreated group, significant improvement of PBF was demonstrated in emodin as well as in sandostatin group which showed reduced PBF following infusion of sandostatin before triggering AHNP. In untreated group plasma TXB was significantly higher, with an increase of 4.5 times, than that in sham-operated group while 6-keto-PGF1 alpha or PGE2 tended to decrease. The above mentioned abnormal synthesis of eicosanoids was blocked either in emodin or in sandostatin group in which lessened damage of acini cells was shown by pathologic scoring or transmission electron microscope. Both of the two groups shared significantly lower mortalities than the untreated group. CONCLUSION: Either emodion or sandostatin could partly reverse the decrease of PBF in the early stage of AHNP, which may be ascribed at least in part to inhibition of abnormal synthesis of eicosanoids and improvement of cytoprotection of acini cells, and combined application of the two drugs might promise positively synergetic action as well. PMID- 9863134 TI - [Advances in the integrated traditional and Western medicine in the treatment of brain edema and intracranial hypertension in children]. PMID- 9863133 TI - [An experimental study on inhibitory effect of Chinese medicine tai-bao on antisperm antibody]. AB - OBJECTIVE: To investigate whether Chinese medicine Tai-bao could inhibit antisperm antibody in experimental mice. METHODS: The experimental immunoinfertility mice were due to antisperm antibody induced by injection of human sperm membrane antigens. The experimental immuno-infertile mice used in the present study were divided into four groups including Tai-bao high dose group (46.8 g.kg-1.d-1), Tai-bao low dose group (31.2 g.kg-1.d-1), prednisone group and normal saline group. The enzyme linked immune sorbent assay (ELISA) and microcytotoxic assay were used for detection of antisperm antibody. The change of levels of antisperm antibody before and after treatment, pregnant rate, and the number of implantation were investigated in tested mice. RESULTS: The pregnant rates in normal saline group, prednisone group, Tai-bao high dose group and low dose were 38.89%, 47.06%, 70.00% and 75.00% respectively. The rate of pregnancy in Tai-bao low dose group was significantly higher as compared with normal saline group (P < 0.05). The rate of implantation in Tai-bao low dose group was significantly higher than that in prednisone group (P < 0.05). The results of detection of the cytotoxic antibody to sperm showed that cytotoxic percentages in Tai-bao high dose group (63.0 +/- 10.3%) and prednisone group (56.3 +/- 13.7%) were significantly lower (P < 0.05 and P < 0.01) than that in normal saline group (72.84 +/- 5.05%). CONCLUSION: Chinese medicine Tai-bao possesses regulatory effect on reproductive immune function, inhibitory effect on antisperm cytotoxic antibody, and promoting effect on pregnancy. PMID- 9863135 TI - [Current status and prospect of research on the treatment of paroxysmal nocturnal hemoglobinuria with integrated traditional Chinese and Western medicine]. PMID- 9863136 TI - Huperzine A (shuangyiping): a promising drug for Alzheimer's disease. AB - Hup A, a novel alkaloid isolated from Chineses herb Huperzia serrata, is a potent and selective inhibitor of AChE, with a rapid absorption and penetration into the brain in experimental animals. The inhibition is reversible with a longer duration of action. Hup A exhibited memory-enhancing activities in a broad range of animal cognitive model. Compared to Phy, Tac, and Gal, Hup A has better therapeutic indices, and peripheral cholinergic side effects are minimal at therapeutic doses. These findings suggest that Hup A is a promising candidate for clinical development as a symptomatic treatment for AD. PMID- 9863137 TI - Chronic treatment with (-)-stepholidine alters density and turnover of D1 and D2 receptors in striatum. AB - AIM: To study the effects of chronic administration of SPD on the density and turnover of striatal D1 and D2 dopamine (DA) receptors. METHODS: Receptor density was monitored by radio-receptor binding assay. The receptor recovery and turnover were studied after irreversible inactivation by N-ethoxycarbonyl-2-ethoxy-1, 2 dihydro-quinoline (EEDQ). RESULTS: Chronic SPD treatment (sc, 20 mg.kg-1.d-1 x 21 d) upregulated both striatal D1 and D2 receptor density. As compared to vehicle treated rats, SPD increased D1 and D2 receptors by 41.5% and 43.7%, respectively SPD also altered the turnover of both D1 and D2 receptors. The degradation rate constant (k = 0.0082.h-1) and the synthesis rate (r = 2.65 pmol.h-1/g protein) of D2 receptors in SPD-treated rats were significantly increased vs vehicle-treated rats (k = 0.0049.h-1; r = 1.10 pmol.h-1/g protein). The degradation rate constant (k = 0.0059.h-1) and the synthesis rate (r = 3.1 pmol.h-1/g protein) of D1 receptors was also increased in SPD-treated rats vs vehicle-treated rats (k = 0.0048.h-1; r = 1.8 pmol.h-1/g protein), but the alteration of degradation rate constant missed significance (P > 0.05). As a result, receptor recovery following EEDQ was accelerated. The half time for D1 and D2 receptors recovery in SPD group were 117.5 h and 84.5 h, respectively, shorter than 144.4 h and 141.4 h in vehicle-treated rats. CONCLUSION: Chronic SPD treatment upregulated D1 and D2 receptors, and accelerated DA receptor turnover and recovery mainly by increasing receptor synthesis. PMID- 9863138 TI - Hypoxia effects on hypothalamic corticotropin-releasing hormone and anterior pituitary cAMP. AB - AIM: To study the effects of acute and chronic hypoxia on hypothalamus-anterior pituitary-adrenocortex axis. METHODS: Rats and pikas were exposed to different altitude and periods. Animals were injected with CRH, Arg and NE in the third ventricle of the brain of rats. RESULTS: Anterior pituitary cAMP and plasma corticosterone levels of rats obviously increased during 1 h of hypoxia. cAMP was increased from 2.23 +/- 0.13 of control group to 7.7 +/- 0.7 of 5 km and 13.4 +/- 1.9 nmol/g wet tissue of 8 km, respectively. i.c.v. CRH, Arg and NE all activated HPA axis. The effects of CRH were most potent. CRH 2 microL 0.75 nmol i.c.v increased anterior pituitary of cAMP from 3.5 +/- 0.4 of control to 22.4 +/- 2.2 nmol/kg wet tissue. Stimulating altitude of 5000 m resulted in a 16.9% decrease in corticosterone level (P < 0.05), 8000 m resulted in a 47.5% decrease (P < 0.01) after hypoxia for 25 d. Hypoxia did not activate HPA axis in pikas. CONCLUSION: 1) Hypoxia stress activates the secretion of corticotrophin (ACTH) via cAMP; 2) Adrenocotical function of rats decays during chronic hypoxia; 3) Arg and NE regulate the secretion of plasma corticosterone and synthesis of pituitary cAMP at the hypothalamus level; 4) Hypoxia tolerance of the pika was high. PMID- 9863139 TI - Ciliary neurotrophic factor antagonizes gentamicin-induced alterations of electric potentials in auditory pathway in guinea pigs. AB - AIM: To study the effects of ciliary neurotrophic factor (CNTF) on the expressions of gentamicin ototoxicity in guinea pigs. METHODS: The auditory function of pigmented guinea pigs was examined using auditory brainstem response (ABR), cochlea microphonic potential (CM), and action potential of auditory nerve (AP). RESULTS: In animals injected gentamicin (80 mg.kg-1.d-1, i.m.), ABR threshold began to elevate on d 20, and prolongations of ABR wave I, IV and the I IV interpeak latencies were observed. The animals treated with gentamicin for 30 d displayed lower amplitudes of CM and AP (N1) than the controls. CNTF (0.44 mg.kg-1.d-1, s.c.) inhibited the gentamicin-induced elevation of ABR thresholds, the prolongation of ABR wave I, IV and the I-IV interpeak latencies, and the decreases in amplitudes of CM and AP (N1). CONCLUSION: CNTF attenuated the gentamicin-elicited auditory impairment in guinea pigs. PMID- 9863141 TI - Effect of procainamide on pulmonary thromboembolism and platelet malondialdehyde in mice. AB - AIM: To study the effect of procainamide (PA) on pulmonary thromboembolism, platelet malondialdehyde (MDA) level and platelet aggregation induced by collagen and adrenaline. METHODS: Pulmonary thromboembolism, 2-thiobarbituric acid fluorescence micro-determination, conventional microscopic counting, and platelet aggregation test were used. RESULTS: PA (10-20 mg.kg-1) and mannitol (200 mg.kg 1) reduced thrombosis by 30%-75% and 75%, respectively. Thrombocytopenia followed thrombosis increased after the pretreatment of PA and mannitol. MDA decreased by both of them in vivo. In vitro, PA inhibited platelet aggregation and MDA production induced concentration-dependently by collagen and adrenaline. CONCLUSION: Inhibition of PA on pulmonary thromboembolism is involved in the decrease of platelet aggregation and MDA production. PMID- 9863140 TI - Modulation by nicotine on binding of cerebral muscarinic receptors with muscarinic agonist and antagonist. AB - AIM: To study the modulatory effects of nicotine on the binding of brain muscarinic receptors. METHODS: The binding of brain muscarinic receptors with the agonist [3H] oxotremorine-M or the antagonist l-[3H]QNB was determined in the presence/absence of nicotine. RESULTS: Pre-incubation of the membrane fraction derived from rat cerebral cortex with nicotine 1.0 mumol.L-1 led to a decrease in the dissociation constant (Kd) for [3H] oxotremorine-M binding to muscarinic receptors, while the maximal binding value (Bmax) was unchanged. The Kd value for binding of the muscarinic antagonist l-[3H]QNB was concentration-dependently increased by preincubation with nicotine 0.1 nmol.L-1-10.0 mumol.L-1, with Bmax unchanged. The effect of nicotine on the Kd for l-[3H]QNB binding was prevented by mecamylamine 10 nmol.L-1, but was enhanced by dithiothreitol 10 mumol.L-1, which by itself was also capable of increasing the Kd value. CONCLUSION: Nicotine increases the affinity of brain muscarinic receptors for muscarinic agonist, but decreases the affinity of brain muscarinic receptors for muscarinic antagonist. PMID- 9863142 TI - Effect of captopril on proliferation of aortic smooth muscle cells. AB - AIM: To study the effect of captopril (Cap) on proliferation of smooth muscle cells (SMC). METHODS: Proliferation of cultured porcine aortic SMC was promoted by hyperlipidemic sera (HLS). RESULTS: Cap (0.2 mg.L-1) remarkably reduced [3H]thymidine ([3H]TdR) incorporation, inhibited DNA synthesis and proliferation of SMC, and lessened morphological changes caused by HLS. After administration of Cap, malondialdehyde (MDA) content and plasminogen activator inhibitor (PAI) activity were reduced, 6-keto-PGF1 alpha and cAMP contents were obviously increased (P < 0.05-0.01). CONCLUSION: Cap inhibited the proliferation of SMC, associated with MDA, PAI, 6-keto-PGF1 alpha and cAMP. PMID- 9863144 TI - Inhibitory effect of potassium channel openers on proliferation of cultured rabbit aortic smooth muscle cells. AB - AIM: To study the effects of pinacidil (Pin), nicorandil (Nic), RP 49356 (RP), and lemakalim (Lem) on phenylephrine (PE)-induced proliferation of cultured rabbit aortic smooth muscle cells (ASMC). METHODS: The [3H]thymidine incorporation into DNA and number of ASMC were measured. RESULTS: [3H]Thymidine uptake induced by PE was markedly inhibited by Pin, Nic, RP, and Lem (0.1-10 mumol.L-1). Only RP inhibited [3H]thymidine uptake without PE-pretreatment. Cell growth induced by PE was completely inhibited by Pin. CONCLUSION: Pin, Nic, RP, and Lem showed potent antiproliferation of ASMC. PMID- 9863143 TI - Effect of chronic captopril treatment on circulating and tissue renin-angiotensin system in SHR rats. AB - AIM: To study the effect of captopril treatment and its withdrawal on the circulating and tissue peptidyl-dipeptidase A, angiotensinogen (AGT), and angiotensin II (A II), in relation to left ventricular hypertrophy (LVH) and systolic blood pressure (SBP). METHODS: SHR male rats were given captopril 100 mg.kg-1.d-1 [SHRcap, number (n) = 43] orally in mixture with milk powder as vehicle from intrautero period of 16 wk of age. Rats were killed at 16 (n = 19) and 40 (n = 24) wk of age, respectively. Male, age-matched untreated SHR and WKY rats served as controls. SBP, left ventricular mass/body weight (LVM/BW) ratio, left ventricular (LV) myocardium and plasma A II concentration, aortic and serum peptidyl-dipeptidase A activity, AGT mRNA level in kidney and liver, renal renin mRNA level were determined. RESULTS: Captopril treatment decreased SBP and reduced LVM/BW at 16 and 40 wk of age, and persistently inhibited LV myocardium A II, aortic peptidyl-dipeptidase A activity, and AGT gene expression in kidney even after the treatment was removed. Nevertheless, no changes were found in plasma A II concentration, serum peptidyl-dipeptidase A activity, and AGT mRNA level in liver by captopril therapy. Renal renin mRNA level was low in SHR and WKY rats, but it was increased by captopril treatment. Tissue renin-angiotensin system (RAS) such as AGT mRNA in kidney, aortic peptidyl-dipeptidase A activity, and LV myocardium A II, rather than circulating RAS (AGT mRNA in liver, renin mRNA in kidney, serum peptidyl-dipeptidase A activity and plasma A II), were persistently inhibited by early captopril treatment, even after the withdrawal of the treatment. CONCLUSION: The long-term inhibition of tissue RAS is one of the mechanisms of the persistent hypotensive effect of captopril treatment. PMID- 9863145 TI - Propranolol and bepridil attenuating levothyroxine-induced rat cardiac hypertrophy and mitochondrial Ca2+ Mg(2+)-ATPase activity elevation. AB - AIM: To study the effects of propranolol and bepridil on levothyroxine-induced rat cardiac hypertrophy and mitochondrial Ca2+ Mg(2+)-ATPase activity elevation. METHODS: Rat heart hypertrophy was induced by i.p., levothyroxine 1 mg.kg-1.d-1 x 10 d. Then rats were treated by ig propranolol (Pro) or bepridil (Bep) 10 mg.kg-1 daily. Ca2+ Mg(2+)-ATPase activity and enzyme kinetic parameters were assayed. RESULTS: The activity and Vmax of mitochondrial Ca2+ Mg(2+)-ATPase isolated from hypertrophic left ventricle were 25 +/- 4 and 35.1 +/- 0.8 mumol Pi.h-1/mg protein, respectively, those of normal were 6.7 +/- 1.8 and 10 +/- 4 mumol Pi.h 1/mg protein, respectively. Apparent K(m) of the hypertrophic group Ca2+ Mg(2+) ATPase was 0.4 +/- 0.12 mmol.L-1 ATP, and that of normal was 0.59 +/- 0.22 mmol.L 1 ATP. The total protein quantity of hypertrophic left ventricle was 80 +/- 30 mg, and that of normal was 47 +/- 9 mg. After treated with Pro or Bep (both 10 mg.kg-1 ig), the cardiac hypertrophy was attenuated, the enzyme activity and Vmax as well as total protein quantity of hypertrophic left ventricle were reduced to normal level, but apparent K(m) was not affected. CONCLUSION: Both Pro and Bep prevented the myocardium and its mitochondria from ischemia and overload calcium injury. PMID- 9863146 TI - Endothelium-dependent effect of perindopril and enalaprilat in rat thoracic aorta. AB - AIM: To study the effect of the angiotensin-converting enzyme (ACE) inhibitors perindopril (Per) and enalaprilat (Ena) on the reactivity of the endothelium in normal rats. METHODS: Male rats were treated intragastrically with Per (2 mg.kg 1.d-1) or placebo (n = 18) for 6 wk. Aorta was isolated for experiment. Another set of isolated aortic rings with and without endothelium were incubated with Ena (0.1 mumol.L-1) for 30 min. Responses to acetylcholine, serotonin, phenylephrine, sodium nitroprusside (SN), and nitroglycerin (Nit) were observed. RESULTS: Endothelium-dependent relaxation to acetylcholine was augmented in aortic rings from rats treated with Per in comparison with control. The IC50 value (95% confidence limits) decreased from 3.8 (0.56-26.1) mumol.L-1 (control group) to 0.98 (0.28-3.41) mumol.L-1 (Per-treated group). The maximal relaxation was augmented from 62 +/- 9% to 78 +/- 10% (P < 0.01). However, the responses to the endothelium-independent vasodilators, SN and Nit, were similar. Serotonin- and phenylephrine-induced contractions were decreased, which were influenced by basal release of endothelium-derived relaxing factor (EDRF). EC50 values was 6.1 (2.6 14.4) nmol.L-1 vs 8.3 (3.6-18.8) nmol.L-1 in comparison with control group and Per-treated group. The maximal contraction was decreased from 2.42 +/- 0.29 g (control group) to 1.96 +/- 0.25 g (treated group) (P < 0.01). Similar results were found in incubation with Ena. CONCLUSION: Ena and Per enhanced the basic release of EDRF from vascular endothelium. PMID- 9863147 TI - Inhibitory effects of Acanthopanax gracilistylus saponins on human platelet aggregation and platelet factor 4 liberation in vitro. AB - AIM: To study the effects of Acanthopanax gracilistylus var pubescens Li saponins (AGVPS) on human platelet aggregation and platelet factor 4 (PF4) liberation in vitro. METHODS: Human platelet aggregations induced by ADP, adrenaline, and collagen were measured turbidimetrically. The aggregation curve was recorded on a platelet aggregometer and the maximal aggregation rate (ARmax), effective deaggregation rate in 5 min (DR5 min) and lag time (LT) were autocalculated by the built-in microcomputer; PF4 liberation from human platelets stimulated by ADP and collagen was determined by recording the heparin thrombin clotting time (HTCT). Thrombosis was tested by weighing the wet and dry thrombi formed in a siliconized revolving ring. RESULTS: AGVPS inhibited in vitro the ARmax with IC50 of 1.33 (95% confidence limits: 1.09-1.63, ADP-induced), 1.66 (1.54-1.79, adrenaline-induced), and 4.2 g.L-1 (0.6-29, collagen-induced). The DR5 min (on ADP-induced aggregation) and LT (collagen-induced) were also increased as well. Meanwhile, AGVPS 0.63-2.50 g.L-1 prolonged HTCT on ADP- and collagen-stimulated PF4 liberation. At 0.34-1.39 g.L-1, AGVPS reduced the wet and dry weight of thrombi formed in vitro. CONCLUSION: AGVPS inhibits human platelet aggregation, liberation, and thrombosis in vitro, suggesting its possible antithrombotic action in man. PMID- 9863148 TI - Synergism of procoagulation effect of thrombin-like enzymes from Dienagkistrodon acutus and Agkistrodon halys snake venoms. AB - AIM: To study the synergistic effect of thrombin-like enzymes (TLE) of Dienagkistrodon acutus (DA) and Agkistrodon halys (AH) venoms. METHODS: TLE were isolated from venoms of DA and AH by successive column chromatography. Effects of combination of DATLE and AHTLE and related factors on the clotting time and clot quality were tested in vitro. RESULTS: One coagulation unit was 2.7 micrograms for DATLE and 304.4 micrograms, for AHTLE with reference standard of thrombin (42.2 micrograms). TLE-induced clot was a fibrin monomer which was fragile and did not retract. Combination of AHTLE and DATLE shortened the clotting time and decreased the solubility of the clot in urea 5 mol.L-1. When subthreshold concentration of thrombin or physiological concentration of Ca2+ was added, the clotting time was further shortened, the clot was no longer soluble in urea and retracted well, and the resistance of the clot to plasmin degradation was increased. CONCLUSION: A synergistic effect of DATLE and AHTLE accelerated hemocoagulation and improve clot quality. PMID- 9863149 TI - Benzylpenicillin induced specific non-IgE antibody response in mice. AB - AIM: To study whether or not the specific non-IgE antibody response in mice can be induced by benzylpenicillin in vivo. METHODS: Antibody response and antigenic cross reactions were determined by enzyme-linked immunosorbent assay (ELISA). Antigen molecules recognized by antibodies were tested by hapten inhibition assay. RESULTS: During d 1-d 50 after immunization, positive % of specific IgM, IgG, and IgA to benzylpenicillin were 100%, 50%-100%, and 17%-100%, respectively. IgM and IgG to benzylpenicillin also recognized ampicillin and piperacillin. The positive % of IgM and IgG to ampicillin were 23%-100% and 50%-100%, to piperacillin 43%-100% and 50%-100%, respectively. Aged benzylpenicillin showed an inhibitory effect on specific antibodies in a dose-dependent manner, and inhibitory % of specific IgM and IgG were 29%-87% and 29%-71%. However, freshly prepared benzylpenicillin had no effect. CONCLUSION: Specific non-IgE antibody response was successfully induced by benzylpenicillin in mice, in which the isotypes were mainly IgM and then IgG and IgA. Antibodies recognized degraded products, not benzylpenicillin molecule itself. Antigenic cross reactions occurred between benzylpenicillin, ampicillin, and piperacillin. Isotypes of antibodies responsible for cross reactions were mainly IgG and then IgM. PMID- 9863150 TI - Tegument changes of Schistosoma japonicum and Schistosoma mansoni in mice treated with artemether. AB - AIM: To study the effect of artemether (Art) on the tegument of schistosomes. METHODS: Mice infected with S japonicum cercariae for 7 and 35 d, or with S mansoni cercariae for 49 d were treated intragastrically with Art 200-300 mg.kg 1.d-1 for 2 d. Schistosomes were collected in groups of 2 mice at various intervals after medication for scanning electron microscopic observation. RESULTS: The tegumental changes induced by Art appeared to be similar in S japonicum and S mansoni: swelling and fusion of tegumental surfaces, vesicle formation and collapse of discoid-like sensory structures. In S japonicum the emergence of tegumental alterations was earlier in 7-d-old schistosomulae than that in 35-d-old adult worms. CONCLUSION: Art injured the teguments of S japonicum and S mansoni. PMID- 9863151 TI - Anti-lipid peroxidation of gomisin J on liver mitochondria and cultured myocardial cells. AB - AIM: To study the influences of gomisin J on lipid peroxidation and calcium paradox. METHODS: Using two in vitro models of rat liver mitochondria membrane lipid peroxidation (LPO) and cultured myocardial cells. RESULTS: Gomisin J inhibited Fe2+/ascorbic acid and ADP/NADPH-induced LPO with IC50 (95% confidence limits) 5.5 (4.5-6.7) and 4.7 (2.8-7.8) mumol.L-1, respectively, when cultured myocardial cells preincubated with Ca(2+)-free medium for 2 min were incubated with normal medium containing Ca2+, a marked increase of malondialdehyde (MDA) formation occurred and gomisin J 10 mumol.L-1 protected myocardial cells through decreasing MDA formation. CONCLUSION: Gomisin J inhibits LPO in rat liver mitochondria and protects cultured myocardial cells from being injured by calcium paradox. PMID- 9863152 TI - Inhibitory effects of quinidine and quinine on liver microsome oxidation enzymes in man and rat. AB - AIM: To compare the inhibitory effects of quinidine and quinine on liver microsome bufuralol 1'-hydroxylase (BH), aryl hydrocarbon hydroxylase (AHH), and 7-ethoxycoumarin O-deethylase (ED) activities in man and rat. METHODS: A normal phase HPLC and the fluorescence spectrometry were used to assay the enzyme activities. RESULTS: Both quinidine and quinine produced a concentration dependent inhibition to liver microsome BH, AHH, and ED in man and rat. Their median inhibitory concentrations (IC50) on liver microsome BH, AHH, and ED low and high affinity phases were 0.2, 378, 2952, and > 5000 mumol.L-1 for quinine in man, 290, 613, 1465, 1595, mumol.L-1 for quinidine in rat; 29, 207, 808, and > 5000 mumol.L-1 for quinine in man, and 31, 54, 597, and 2508 mumol.L-1 for quinine in rat, respectively. CONCLUSION: Quinidine is a species- and stereo selective potent inhibitor to human liver microsome BH. PMID- 9863153 TI - Resistance to apoptosis of harringtonine-resistant HL60 cells induced by tetrandrine. AB - AIM: To study the mechanism of resistance to apoptosis in the harringtonine (Har) resistant HL60 cells with tetrandrine (Tet). METHODS: Growth inhibition, flow cytometry, DNA agarose gel electrophoresis, protein phosphorylation, and RNA dot hybridization. RESULTS: The resistant cells had no cross resistance to Tet. Tet induced the sensitive but not the Har-resistant HL60 cells to apoptosis. The high phosphorylation of protein < 30 kDa occurred when the resistant cells were treated with Tet. Tet and Har increased the expression of c-myc mRNA in the sensitive HL60 cells. The expression of c-myc mRNA in the resistant cells was obviously decreased and almost not changed in treatment with Tet and Har. CONCLUSION: Tet induced the sensitive but not the Har-resistant HL60 cells to apoptosis, and the resistance to apoptosis induced by Tet was associated with the high protein phosphorylation and reduction of the expression of c-myc mRNA. PMID- 9863154 TI - [Effect of magnesium sulfate on calcium channels in isolated cardiomyocytes of guinea pig]. PMID- 9863155 TI - Patch clamp study on mechanism of adenosine-induced inhibitory effects in frog pituitary melanotrophs. AB - Our laboratory demonstrated that adenosine inhibits the activation of adenylyl cyclase and the secretion of the alpha-melanocyte-stimulating hormone (alpha-MSH) from the intermediate lobe of the frog pituitary. This paper showed the bioelectric effects induced by adenosine, the ionic conductances modulated by adenosine, and the possible involvement of intracellular messengers, indicated the mechanism by which adenosine controls the secretion of alpha-MSH. The results show that adenosine acting on A1 adenosine receptor subtype reduced the Ca2+ influx necessary for the secretion, through 4 distinct mechanisms: 1) a hyperpolarization resulting from the activation of a voltage-insensitive K+ conductance, 2) a reduction of the duration of spontaneous action potentials due to an increase of the outward delayed rectifyer K+ current (lk), 3) a diminution of the cellular excitability by an activation of the transient outward K+ current (lA), and 4) an inhibition of the L- and N-type Ca2+ currents, with a predominant action on the N-type component. Cell dialysis with GTP gamma S rendered irreversible the effects of adenosine on the K+ conductances and Ca2+ channels, whereas PTX pretreatment totally abolished the response to adenosine, suggesting all bioelectric effects of adenosine were mediated by pertussis toxin-sensitive G proteins. Whether the implicated G proteins regulate the K+ and Ca2+ channels by tight-coupling or via a second-messenger system remains to be solved. With our results, the involvement of adenylyl cyclase can be excluded because addition of cAMP and IBMX, an inhibitor of phosphodiesterases, in the intracellular solution, or application of dibutyryl cAMP in the extracellular solution did not modify the adenosine-induced responses. PMID- 9863157 TI - Modification of an ion-paired high pressure liquid chromatography for ticarcillin in human serum. AB - AIM: To develop a new HPLC assay for the clinical pharmacokinetic study of ticarcillin in human. METHODS: Reverse phase ion-paired HPLC was developed with a Novapak C18 column. The mobile phase consisted of 20% of acetonitrile and 80% of phosphate buffer 10 mmol.L-1 (pH 2.0-2.5) containing tetrapentylammonium bromide of 1 mmol.L-1. The analysis was monitored at 225 nm and the flow rate of the mobile phase was 0.8 mL.min-1. Cefoxitin was used as an internal standard. The sample volume was 200 microL of serum. RESULTS: The assay was linear in the range of 2-80 mg.L-1. The intrarun coefficients of variation (CV) for a ticarcillin 5 mg.L-1 check sample was 4.2% (n = 10), and 2.7% for a 60 mg.L-1 check sample (n = 10), respectively. The inter-run CV (n = 6) were 1.6% (5 mg.L-1) and 2.3% (60 mg.L-1), respectively. The average recovery of the assay was 96.6%. CONCLUSION: This assay was sensitive, precise, and accurate for evaluating the clinical pharmacokinetics of ticarcillin. PMID- 9863156 TI - Oxytocin-stimulated insulin release in a clonal beta-cell line RINm5F: involvement of phospholipase C-dependent and -independent pathways. AB - AIM: To study the mechanisms underlying oxytocin (Oxy)-induced insulin release. METHODS: In a clonal pancreatic beta-cell line, RINm5F cells. RESULTS: Oxy increased insulin release and [Ca2+]i in a concentration-dependent manner. Oxy induced insulin release was not altered by pretreatment with pertussis toxin (PT). U-73122 (2-8 mumol.L-1), a phospholipase C (PLC) inhibitor, concentration dependently inhibited Oxy-induced increases in [Ca2+]i with IC50 value of 2.8 +/- 0.2 mumol.L-1. In addition, U-73122 diminished the Oxy-induced increase in intracellular concentration of inositol 1, 4, 5-triphosphate (IP3). U-73122 at 8 mumol.L-1 totally abolished the Oxy-induced increases in [Ca2+]i and IP3; however it reduced the Oxy-induced increase in insulin release only by 36% and 63% in the monolayer and suspended cell preparations, respectively. CONCLUSION: Oxy increases insulin release through both PLC and non-PLC mediated signal transduction mechanisms. PMID- 9863158 TI - Inhibition of imipramine N-demethylation by fluvoxamine in Chinese young men. AB - AIM: To determine effect of fluvoxamine (Flu) on imipramine (lmi) N-demethylation in healthy Chinese subjects. METHODS: The pharmaco-kinetic parameters of a single oral dose of lmi (100 mg) were compared before and during 2 wk coadministration of Flu (100 mg.d-1). RESULTS: Flu resulted in a doubling of Cmax (112 +/- 33 vs 62 +/- 22 micrograms.L-1, P < 0.01) and an increase in AUC0- infinity (3.9 +/- 1.6 vs 1.5 +/- 0.6 mg.h.L-1, P < 0.01) of lmi. The T1/2 of lmi was prolonged (39 +/- 8 vs 23 +/- 7 h, P < 0.01) due to a marked reduction in Cl0 of lmi (0.6 +/- 0.3 vs 1.3 +/- 0.6 L.h-1.kg-1, P < 0.01). The Cmax of N-demethylated metabolite desipramine (Des) of lmi was decreased (10 +/- 2 vs 18 +/- 5 micrograms.L-1, P < 0.01) and AUC0- infinity of Des was reduced by 54% (0.61 +/- 0.20 vs 1.3 +/- 0.5 mg.h.L-1, P < 0.01) during coadministration with Flu. CONCLUSION: Flu can markedly inhibit lmi N-demethylation in Chinese young men. PMID- 9863159 TI - Doxorubicin cellular pharmacokinetics plays no role in chemosensitizing effect of verapamil on Swiss-3T3 cells. AB - AIM: To find whether or not the doxorubicin (Dox) cellular pharmacokinetics plays a role in chemosensitizing effect of verapamil (Ver) on drug sensitive cells. METHODS: Cytotoxicity and cellular Dox contents (during accumulation and retention periods) were measured in the absence and presence of verapamil in Swiss-3T3 cells and compared with those in multidrug resistant (MDR) MCF-7Adr cells and drug sensitive MCF-7WT cells. mdr-1 mRNA expression in Swiss-3T3 cells was analyzed. RESULT: Dox cytotoxicity was enhanced 2.0-fold in Swiss-3T3 cells by Ver (3 mumol.L-1) and 3.6-fold in MCF-7Adr cells by Ver (6 mumol.L-1), but not in MCF-7WT cells (Ver 6 mumol.L-1). Cellular accumulation of equi-effective concentrations of Dox increased at 6-h incubation in the presence of Ver in Swiss 3T3 (1.5-fold)i and MCF-7WT cells (2.1-fold) but decreased rapidly in MCF-7Adr cells by 20% to 50% compared to that in the absence of Ver. Cellular retention of Dox decreased after 10-min increase in the presence of Ver in Swiss-3T3 cells compared to that in the absence of Ver, that was similar to that in MCF-7WT cells, while the retention was augmented by Ver in MCF-7Adr cells. Slot blot analysis of RNA revealed no mdr-1 gene expression in Swiss-3T3 cells. CONCLUSION: Changes in cellular accumulation and retention of Dox did not account for the chemosensitizing effect of Ver on Swiss-3T3 cells. PMID- 9863160 TI - Pharmacokinetics of weekly transdermal estradiol controlled delivery system in postmenopausal Chinese. AB - AIM: To study Pharmacokinetical profile of weekly transdermal estradiol controlled delivery system (E2-WTCDS) in 18 Chinese postmenopausal women. METHODS: Single and multiple daily doses of E2-WTCDS were applied to an area of intact skin on the lower abdomen, using Estraderm TTS (E2-TTS) from Ciba-Geigy Company as control. E2 concentration in serum was measured by RIA. RESULTS: Cmax 38 +/- 5 ng.L-1, Tmax 22 +/- 5 h, AUC 5651 +/- 386 ng.h.L-1, Cmax(SS) 44 +/- 7 ng.L-1, Cmax(SS) 35 +/- 4 ng.L-1, Fl 0.22 +/- 0.10. The relative bioavailability of E2-WTCDS was comparable to E2-TTS during the same period of system application. CONCLUSION: E2-WTCDS maintained relatively constant and effective serum estradiol concentrations and is suitable for once-weekly application. PMID- 9863161 TI - Diltiazem concentrations in plasma vs PR intervals on electrocardiogram in 8 men. AB - AIM: To develop an acute tolerant model in describing relationship between diltiazem (Dil) concentrations in plasma and PR intervals on ECG in men. METHOD: Both plasma concentrations of Dil and changes of ECG were simultaneously determined after po Dil 90 mg in 8 men. RESULTS: A two-compartmental pharmacokinetic model with first-order input gave a good fitting for the plasma concentration of Dil. Corresponding pharmacokinetic parameters were estimated: t1/2 beta, 5.9 +/- 1.0 h; MRT, 15.9 +/- 1.0 h; t0, 0.38 +/- 0.07 h; tmax, 2.7 +/- 0.4 h, and Cmax, 161 +/- 60 micrograms.L-1. The good fittings for plasma concentration-effect data were obtained with tolerant model E = S x C/(1 + T/T50). The pharmacodynamic parameters were given as follows: S, 829 +/- 293 s.g 1.L; Kt0, 0.037 +/- 0.024 h-1 and T50, 10 +/- 4 micrograms.L-1. CONCLUSION: Relationship between Dil concentrations in plasma and PR interval changes in men after po 90 mg was described using an acute tolerant model. PMID- 9863162 TI - Pharmacokinetics of germanium after po beta-carboxyethylgermanium sesquioxide in 24 Chinese volunteers. AB - AIM: To compare the pharmacokinetics after po different doses of beta carboxyethylgermanium sesquioxide (Ge-132). METHODS: An atomic absorption spectrophotometric system was used to measure germanium concentrations in plasma and urine samples after po Ge-132 1 (low dose, LD), 2.5 (medium dose, MD), and 4 (high dose, HD) g.m-2 in 24 healthy volunteers (one dose per 8 subjects). RESULTS: T1/2 alpha (LD, 1.2 +/- 0.7 h; MD, 1.1 +/- 0.6 h; HD, 1.2 +/- 0.5 h), T1/2 beta (LD, 5.2 +/- 1.2 h; MD, 5.8 +/- 2.5 h; HD, 5.5 +/- 1.4 h) and Cl/F (LD, 33 +/- 12 L.h-1; MD, 35 +/- 10 L.h-1; HD, 33 +/- 11 L.h-1) were not dose-related. Tmax was between 0.75 h and 2 h. Cmax (LD, 5.3 +/- 2.2 mg.L-1; MD, 13 +/- 5 mg.L 1; HD 18 +/- 8 mg.L-1, HD) and AUC (LD, 31 +/- 13 mg.h.L-1; MD, 60 +/- 16 mg.h.L 1; HD, 79 +/- 42 mg.h.L-1) were positive correlation to the dose of Ge-132. Urine eliminated germanium within 24 h accounted for 11 +/- 3% of LD, 9 +/- 3% of MD, and 6 +/- 5% of HD (calculated from Ge/F) and showed a negative correlation to the dose. CONCLUSION: 1) Intracorporal process of Ge after po Ge-132 coincided with the first-order absorption and elimination with two-compartment kinetic model; 2) The amount of germanium eliminated in urine was below 11%. PMID- 9863163 TI - Inhibitory effects of dextromethorphan on c-fos protein expression during focal cerebral ischemia in rats. AB - AIM: To study the effect of dextromethorphan (DM) in focal cerebral ischemia. METHODS: The c-fos protein was detected immunohistochemically in the brain of rats after focal cerebral ischemia (induced by placing a nylon thread in the lumen of the internal carotid artery) with and without treatment with DM. RESULTS: Focal cerebral ischemia induced c-fos protein expression outside the core territory of the middle cerebral artery (MCA) and neuronal damage in the core territory of the MCA. There was an evident expression of c-fos protein in the ipsilateral regions outside the MCA territory (e.g. cingulate cortices, piriform cortices and entorhinal cortices), and in the contralateral regions of hippocampus after 4-h reperfusion following 1-h MCA occlusion. But morphological results showed severe edema and neuronal damage in the core territory and the ipsilateral hippocampus. DM blocked both the c-fos protein induction and neuronal damage in all regions. CONCLUSION: DM reduced c-fos protein expression and blocked the neuronal damage after focal cerebral ischemia. PMID- 9863164 TI - Analgesic activity and selectivity for opioid receptors of enantiomers of ohmefentanyl. AB - AIM: To study analgesic activity and selectivity for opioid receptor subtypes of 8 enantomers of ohmefentanyl (Ohm). METHODS: Analgesic activity was evaluated using the hot plate test in mice. Selectivity for opioid receptors was determined in binding assay and bioassay. RESULTS: Among the 8 enantiomers of Ohm, the most potent isomer was F-9204, (+)-cis-(3R, 4S, 2'S)-Ohm, with an analgesic ED50 (i.p.) value of 1.1 (0.8-1.3) micrograms.kg-1. These enantiomers selectively acted on mu opioid receptors. Ki values (mu) of enantiomers F-9204 and F-9202, ( )-cis-(3R, 4S, 2'R)-Ohm were 4.0 +/- 2.0 and 5 +/- 4 pmol.L-1, respectively. Their Ki(delta)/Ki(mu) ratios were 22,500 and 22,800, respectively. On guinea pig ileum and mouse vas deferens F-9202, F-9204, F-9205, F-9206, F-9207, and F-9208 exhibited very potent inhibitons, which were antagonized by naloxone. In rabbit vas deferens these enantiomers had no effect. CONCLUSION: Eight enantiomers of Ohm selectively acted on mu opioid receptors. F-9204 showed the strongest analgesic activity and the highest selectivity for mu opioid receptors. PMID- 9863165 TI - Direct evidence for histamine H3 receptor-mediated inhibition of norepinephrine release from sympathetic terminals of guinea pig myocardium. AB - AIM: To study the histamine H3 receptors mediated inhibition of norepinephrine (NE) release from cardiac sympathetic terminals of guinea pig isolated atria. METHODS: Release of NE induced by electric field stimulation (50 mA, 5 ms) in the bath solution was measured by HPLC-ECD. RESULTS: The release of NE caused by field stimulation was attenuated by (R)-alpha-methyl-histamine (alpha-MeHA, 0.1 nmol.L-1(-10) mumol.L-1) in a concentration-dependent manner. Thioperamide concentration-dependently antagonized the inhibition of alpha-MeHA. Blockade of H1, H2, alpha 2, beta 2-receptors failed to prevent the inhibitory effect of alpha-MeHA. Thioperamide (1 nmol.L-1(-10) mumol.L-1), when used alone, concentration-dependently facilitated the release of NE evoked by field stimulation. CONCLUSION: The presynaptic histamine H3-receptors inhibited the NE release from cardiac sympathetic terminals. PMID- 9863166 TI - Effects of HI-6 on muscle acetylcholine receptor: analysis on minimal reaction model. AB - AIM: To study the action of 1-(2-hydroxyiminomethyl-1-pyridino)-3-(4-carbamoyl-1 pyridino)-2-oxapropane dichloride (HI-6) on skeletal muscle acetylcholine receptor (N2-ChR). METHODS: N2-ChR was expressed in Xenopus laevis oocyte after injection with mRNA extracted from denervated rat leg muscles. The inward membrane currents induced by various concentrations of carbamylcholine and effects of HI-6 or d-tubocurarine on the currents were measured with voltage clamp technique by fast cell flow of agents. The actions of HI-6 and d tubocurarine on N2-ChR were analyzed by using the minimal reaction model. RESULTS: K of 40.05, 156.00, and 334.67 mumol.L-1 for HI-6, K of 0.02, 0.10, and 0.18 mumol.L-1 for d-tubocurarine were obtained by using the competing for single acetylcholine (ACh)-binding site model, the competing for two ACh-binding sites model, and the noncompetitive inhibition model, respectively. CONCLUSION: HI-6 and d-tubocurarine competed for two ACh-binding sites of N2-ChR with equal affinity to antagonize the effects of the agonist on N2-ChR. The N2-ChR inhibition by HI-6 is much weaker than that by d-tubocurarine. PMID- 9863167 TI - Bepridil reducing levothyroxine-induced enhancement of mitochondria Ca2+ Mg(2+) ATPase activity in rat cerebrum. AB - AIM: To study if bepridil (Bep) could affect the enhancement of activity of cerebral mitochondria Ca2+ Mg(2+)-ATPase caused by levothyroxine (Lev) in relation to ischemic overload calcium cerebrum injury. METHODS: The experimental hyperthyroidism model with ischemic cerebrum was developed in rats by ig Lev 1 mg.kg-1.d-1 for 7 d. Ca2+ Mg(2+)-ATPase activity and its kinetic parameters were assayed. RESULTS: The activity, Vmax and Km of cerebral mitochondria Ca2+ Mg(2+) ATPase in control rats were 3.1 +/- 0.8, 5.1 +/- 2.3 mmol.P(i).h-1/g protein and 0.81 +/- 0.08 mmol.L-1 (ATP) respectively, whereas those of hyperthyroid rats were significantly altered to 4.6 +/- 0.5, 8.5 +/- 1.9 mmol.P(i).h-1/g protein and 0.49 +/- 0.11 mmol.L-1 (ATP) respectively. After treated with Bep 10 or 20 mg.kg-1.d-1 ig for 3 d, allabove 3 parameters of the enzyme were very significantly reduced vs those of either control or hyperthyroid. CONCLUSION: Bep, via decreasing Ca2+ Mg(2+)-ATPase activity and increasing the affinity of Ca2+ Mg(2+)-ATPase to ATP, could prevent rat cerebrum from ATP depletion and ischemic overload calcium injury. PMID- 9863168 TI - Effects of dopexamine on heart function of isolated hypoxic rabbit heart and in comparison with fenoldopam and procaterol. AB - AIM: To study the effects of dopexamine hydrochloride (Dop) on heart functions and coronary flow (CF) of normal and hypoxic isolated perfusing heart and compare the effects of Dop with those of fenoldopam hydrochloride (Fen) and procaterol hydrochloride (Pro). METHODS: The isolated rabbit normal and hypoxic hearts were perfused with Krebs-Henseleit (K-H) solution via aorta at 37 +/- 0.5 degree C. The drug were added into the K-H solution. RESULTS: Dop increased the CF, left ventricular contractile function (+ dp/dtmax etc) and heart rate (HR) in normal heart; Fen increased the heart function more potently, but increased the CF to a lesser degree than Dop did, whereas the effects of Pro was the least. In simple hypoxic group, at 30 min of perfusion, the + dp/dtmax and CF decreased by 66 +/- 4% and 48.1 +/- 1.0%, respectively. Dop remarkably attenuated the decreases in both CF and heart function during hypoxia as it decreased the + dp/dtmax by 32.0 +/- 2.4% and CF by 28 +/- 3%. Fen was less potent than Dop in attenuating the diminution of CF, while Pro was the least effective. CONCLUSION: Dop has a better prospect in the treatment of ischemic heart disease in view of its dual action of increasing CF and inotropism. PMID- 9863169 TI - Effects of sodium pentobarbital on electric and mechanical activities of guinea pig papillary muscle. AB - AIM: To study the effects of sodium pentobarbital (SP) on the action potential (AP) and contraction of guinea pig papillary muscle. METHODS: Using conventional glass microelectrode and mechanical recording of myocardium contraction. RESULTS: SP (> or = 10 mumol.L-1) prolonged the AP duration (APD) and effective refractory period (ERP), while amplitude (APA) and Vmax of phase 0 showed no changes. The effects of SP were abolished by pretreatment with cromakalim, an agonist of ATP sensitive K+ channel. The maximal isometric tension (Pmax) and velocity of tension development (dT/dt) were decreased to 51% and 48% of control, respectively. The first postrest beat (B1) and second postrest beat (B2) were also depressed. CONCLUSION: SP affected the action potential by reducing activities of the K+ channels and reduced the contraction of guinea pig myocardium. PMID- 9863170 TI - Effects of quercetin on production and release of endothelin and cGMP from cultured endothelial cells. AB - AIM: To evaluate the effects of quercetin (Que) on the formation and release of endothelin (ET) and cyclic guanosine monophosphate (cGMP) by endothelial cells. METHODS: Radioimmunoassay (RIA) was used to assess the amounts of ET and cGMP produced by unstimulated human umbilical vein cells (HUVEC) or HUVEC stimulated with activated platelets and thrombin. RESULTS: Following the treatment of the cultured HUVEC with activated platelets, the amount of ET in the medium was increased, but no significant effect on cGMP within the cells was seen. Thrombin not only increased the amount of ET in the medium, but also raised the level of cGMP within the cells. Pretreatment of HUVEC with Que 0.5, 5, and 50 mumol.L-1, the release of ET by HUVEC stimulated with thrombin and thrombin-activated platelets was much inhibited. Que 0.5-50 mumol.L-1 markedly increased the amount of cGMP in HUVEC stimulated with thrombin and activated platelets. Que also inhibited the release of ET from unstimulated HUVEC, but increased the production of cGMP from unstimulated EC. CONCLUSION: Thrombin and activated platelets stimulated the release of ET from HUVEC and Que inhibited the release of ET and promoted the formation of cGMP from HUVEC. PMID- 9863171 TI - Ischemic preconditioning mediated by calcitonin gene-related peptide in isolated rat hearts. AB - AIM: To study the mediating effect of calcitonin gene-related peptide (CGRP) in ischemic preconditioning in the isolated perfused rat heart. METHODS: Isolated rat hearts were subjected to 3 cycles of a 5-min ischemia and a 5-min reperfusion before a 30-min global ischemia followed by a 30-min reperfusion. RESULTS: Ischemic preconditioning caused an improvement of heart functions, reduced the incidence of ventricular arrhythmias and decreased the release of creatine kinase (CK) during reperfusion (CK activities = 0.30 +/- 0.07, 2.03 +/- 0.49, and 0.92 +/- 0.40 U.min-1/g wet wt for control, ischemia-reperfusion, and preconditioning, respectively). However, pretreatment with CGRP8-37 (0.1 mumol.L-1) abolished the improvement of cardiac contractivity, the reduction of the incidence of arrhythmias, and the inhibition of CK release by preconditioning (CK activities = 0.9 +/- 0.4 vs 2.55 +/- 0.32 U.min-1/g wet wt, P < 0.01). CONCLUSION: CGRP is an endogenous myocardial protective substance that played an important role in mediation of ischemic preconditioning. PMID- 9863172 TI - Effect of captopril on intracellular pH in vascular smooth muscle cells. AB - AIM: To observe the effect of captopril (Cap) on intracellular pH (pHi) in aortic smooth muscle cells (ASMC). METHODS: Cultured ASMC derived from rat and rabbit aortae were loaded with the fluorescent dye BCECF and pHi was determined using digital image processing method. RESULTS: The pHi of untreated SHR and WKY rats were 7.37 +/- 0.29 and 7.19 +/- 0.31, respectively. Oral Cap decreased pHi (7.11 +/- 0.26, P < 0.05) and exaggerated pHi response to angiotensin II (Ang-II, 0.1 mumol.L-1) in ASMC of SHR rats vs WKY rats (0.14 +/- 0.05 vs 0.21 +/- 0.05 pH units, P < 0.01). Cap in vitro had no effect on Ang-II induced intracellular alkalinization in ASMC of rabbits. CONCLUSION: Oral Cap inhibits Na+(-)H+ exchange activity in ASMC of SHR rats. PMID- 9863173 TI - Assay of ditiocarb sodium and its methyl ester in mouse plasma by column switching HPLC. AB - AIM: To establish a column-switching high pressure liquid chromatograpy (CSHPLC) with direct injection for determination of ditiocarb sodium (DTC) and its methyl ester (DTC-Me) in mouse plasma. METHODS: After automated online pretreating column enrichment and clear-up, DTC-Me was flushed and chromatographed on an ordinary reversed-phase analytical column, and monitored by UV at lambda absorption = 276 nm. DTC was methylated before determination. RESULTS: No potential interfering peaks were identified. The calibration curves of both analytes were linear within the range of 0.1-150 mg.L-1. The correlation coefficients of DTC and DTC-Me were 0.9998 and 0.9995, respectively. The detection limit was 25 micrograms.L-1 and the coefficient of variation in the assay was within 7% for both compounds. The average recoveries were in the range of 95.28 -100.06%. A typical application was presented for mouse after i.v. DTC 50 mg.kg-1. CONCLUSION: The rapid CSHPLC method with direct injection can be used for the study of pharmacokinetics of DTC and DTC-Me. PMID- 9863174 TI - Synergistic effect of counterregulatory hormones during insulin-induced hypoglycemia in rats: participation of lipolysis and gluconeogenesis to hyperglycemia. AB - AIM: To study the synergistic effect of G (glucagon, 0.02 mg.kg-1), H (hydrocortisone, 20 mg.kg-1) and E (phenylephrine + isoproterenol, both 1 mg.kg 1) during insulin-induced hypoglycemia (IIH) in rats with 6 h of food deprivation (F6 group). METHODS: I (insulin, 1 U.kg-1) was injected i.p. and 30 min later saline (F6 + I group), H, G and E individually or combined (G + H, G + E, H + E and G + H + E) were all injected i.p. and all experiments started 1 h after I injection. RESULTS: The rise in glycemia with H + G + E was greater than the sum of the responses to i.p. H, G and E individually or in double combination plus any single hormone. This effect was reproduced by G + H + Iso (isoproterenol, 1 mg.kg-1), G + H + Iso + Met (metoprolol, 1 mg.kg-1) and G + H + Sal (salbutamol, 1 mg.kg-1). A clear relationship was shown between glycemia and free fatty acids levels. Liver gluconeogenesis from glycerol (2 mmol.L-1) was higher in the group which received G + H + beta-adrenergic agonist vs control rats (F6 or F6 + I groups). CONCLUSION: (a) Acute hyperglycemia is obtained from a condition of IIH by combined i.p. of G + H + beta-adrenergic agonists; (b) This effect cannot be ascribed to a single hormone, but is a consequence of the combined effects of these substances; (c) Blood insulin levels and liver glycogen have no participation; (d) Lipolysis mediated by a beta-adrenergic mechanism and gluconeogenesis from glycerol contribute to the hyperglycemia. PMID- 9863175 TI - Exacerbation of cold restraint-induced gastric ulcer by GABA in mice. AB - AIM: To study the effect of GABA on cold restraint-induced gastric ulcer (CRGU) and its mechanism. METHODS: CRGU was produced in adult male mice at 1 degree C in cold room under restraint for 1 h. The ulcer index indicated the severity of gastric mucosa lesion. Gastric mucin was determined with alcian blue dye. RESULTS: GABA 2 mumol (i.c.v.) accelerated CRGU, Bicuculline (Bic) 0.2 mumol (i.c.v.) did not affect CRGU, whereas Baclofen (Bac) 4 mumol (i.c.v.) exacerbated CRGU. Bic did not modify the exacerbation of CRGU by GABA and Bac. Atropine (Atr) 0.2 mg.kg-1 (s.c.) and Phentolamine (Phen) 2.5 mg.kg-1 inhibited CRGU, and abolished the exacerbation of CRGU by GABA. Cold restraint (CR) decreased the amount of the gastric mucin, but GABA 2 mumol had no effect on CR gastric mucin. CONCLUSION: While GABA-B receptor in brain was activated, exogenous GABA exacerbated CRGU via vagal and sympathetic nerves, bearing no relation to decrease of the gastric mucin or weakening of the gastric mucosal barrier. PMID- 9863176 TI - Characteristics of harringtonine-resistant human leukemia HL60 cell. AB - AIM: To study the mechanisms of the resistance to harringtonine (Har) in the HL60 cells. METHODS: Growth inhibition, karyotype analysis, flow cytometry, Western blotting and polymerase chain reaction. RESULTS: The Har-resistant HL60 cell line, named HR20, showed cross resistance to homoharringtonine, doxorubicin, daunorubicin, vincristine, and colchicine. The growth doubling time and the cell numbers in G1 phase were increased. The accumulation of cellular daunorubicin in the resistant cells was obviously reduced, but distinctly increased by tetrandrine and verapamil. The numbers of telocentromeric chromosome increased and the chromosomal aberration more occured in the resistant cells. The resistant cells overexpressed multidrug resistant mdr-1 gene and P-glycoprotein 150 kDa. CONCLUSION: The Har-resistant HL60 cell strain belonged to a multidrug resistance strain, overexpressing mdr-1 gene and P-glycoprotein. PMID- 9863177 TI - Oxidized low-density lipoproteins induce apoptosis in macrophages. AB - AIM: To examine whether oxidized low density lipoproteins (ox-LDL) might induce apoptosis in mouse peritoneal macrophages (MPM). METHODS: Low density lipoproteins (LDL) were isolated from healthy human plasma by ultracentrifugation and oxidized by CuSO4 10 mumol.L-1. MPM were incubated in a medium containing ox LDL, LDL, or phosphate-buffer solution (PBS) as control. DNA fragmentation was visualized by agarose gel electrophoresis and determined quantitatively using Hoechst 33,258 fluorochrome. RESULTS: Ox-LDL, not LDL, elicited typical apoptotic morphological changes (shrinkage of cytoplasm and condensation of chromatin) and DNA fragmentation in a time- and dose-dependent manner. Incubation for 24 h was necessary for ox-LDL 200 mg protein.L-1 to induce DNA fragmentation, and the maximal effect reached at 72 h. The DNA fragmentation after 24 h incubation with ox-LDL at concentrations of 100, 200, and 400 mg protein.L-1 amounted to 6.0% (P > 0.05), 9.3% (P < 0.05), and 30.9% (P < 0.01), respectively vs PBS control. Dextran sulfate, a scavenger receptor blocker and cycloheximide, a protein synthesis inhibitor, exhibited no effect on DNA fragmentation. However, antioxidant butylated hydroxytoluene (BHT) 20 mumol.L-1 completely inhibited Cu(2+)-mediated oxidation of LDL as well as the apoptosis-inducing effect of Cu(2+)-exposed LDL. Lysophosphatidylcholine (LPC), an active component in ox-LDL, at concentration up to 60 mumol.L-1, did not elicit DNA fragmentation in MPM. CONCLUSION: Ox-LDL induces apoptosis in MPM without involving LPC. PMID- 9863178 TI - Correlation between expression of mdr-1 gene and oncogenes in human promyelocytic leukemic HL60 cell line and sublines. AB - AIM: To study the relationship between expression of oncogenes and multiple drug resistant (MDR) phenotype. METHODS: The drug resistant level of HL60 cell line and its sublines were determined with flow cytometry. RNA Dot blot hybridization was used to identify the expression of oncogenes and mdr-1 gene. RESULTS: The expression of mdr-1 gene was in the opposite relation with c-myc expression, but in the positive relation with c-H-ras gene expression in the multiple drug resistant cell lines. In non-MDR cell line HL60/RA, the expression levels of mdr 1, c-myc, and c-H-ras were the same as HL60 parental cells. CONCLUSION: Multiple drug resistance is related to not only mdr-1 expression, but also some oncogenes expression level. PMID- 9863179 TI - [DNA damage induced by fotemustine in cultured HL60 cells]. AB - AIM: To detect DNA damage caused by fotemustine (Fot). METHODS: DNA damage in HL60 cells was evaluated by modified alkaline elution technique. DNA interstrand crosslink (ISC) and DNA-protein crosslink (DPC) were determined. Carmustine (Car) was used as control. Drug treatment time was 1 h. RESULTS: After treatment with Fot 300 mumol.L-1, ISC and DPC index were 5.4 and 6.1 at 6 h, 2.7 and 1.2 at 12 h, respectively. ISC reached the maximum at about 6 h. For Fot 100 mumol.L-1, ISC and DPC index at ISC peak time were 2.1 +/- 0.9 and 3.55 +/- 0.23, 5.0 +/- 0.5 and 7.7 +/- 1.1 for Car 100 mumol.L-1, respectively. Single strand break (SSB) was induced by Fot. CONCLUSION: Fot caused HL60 cell DNA ISC, DPC, and SSB. ISC was formed more quickly by Fot than that by Car. PMID- 9863180 TI - [Blockade of (+/-) 12-chloroscoulerine on feed-back regulation of dopamine D2 autoreceptors]. AB - AIM: To verify whether (+/-) 12-chloroscoulerine (CSL) is antagonist or agonist effect to D2 autoreceptors. METHODS: The levodopa content accumulated in the rat striatum was measured by HPLC-ECD, and the DA neuron firing activity in the substantia nigra zona compacta (SNC) was recorded. RESULTS: The accumulated levodopa content induced by CSL 40 mg.kg-1 was much more than that of 1,4-butyro lactone (BL) group (P < 0.01). After i.p. injection of apomorphine (Apo) 5 mg.kg 1, the levodopa content was decreased below that of BL group (P < 0.05). The Apo inhibition on levodopa content was completely reversed by CSL (40 mg.kg-1, i.p.) and then increased the levodopa content (2.5 +/- 1.1 micrograms.g-1) over that of Apo group (0.7 +/- 0.3 microgram.g-1, P < 0.01). In the electrophysiologic recording, Apo (15 micrograms.kg-1, i.v.) induced the decrease of SNC DA cell firing rate nearly to zero. At the accumulated dose of CSL up to 80 micrograms.kg 1 (i.v.), the inhibition of Apo was attenuated and the firing activity was restored to predrug level. CONCLUSION: CSL showed an antagonistic action, an action to D2 autoreceptors. PMID- 9863181 TI - [Effects of ACh on the electric activity of rostral ventrolateral medullary neurons of rat in vitro]. AB - Extracellular single-unit discharges were obtained from 165 spontaneously active neurons within the region of the rostral ventrolateral medulla (RVLM) by glass microelectrode from 89 brain slices of the Sprague-Dawley rats. The units could be divided into three types: regular (61.8%), irregular (24.2%) and silent (14%). Acetylcholine (ACh, 0.1, 0.3 mumol/L) showed four kinds of effects on spontaneous discharges of RVLM neurons: excitatory, inhibitory, biphasic and non-responsive, counting respectively 41.8%, 20%, 3% and 35.2% of the neurons tested. The excitatory effect of ACh was dose-dependent. The effects, either excitatory or inhibitory, of ACh (n = 49) were mostly blocked by atropine (0.3 mumol/L, n = 42). The excitatory effect of ACh (n = 14) could be blocked mainly by selective antagonist of M1 receptor, pirenzepine (PZ, 30 nmol/L, n = 9), but not by selective antagonist of M2 receptor, methoctramine (MT) and AFDX-116. The inhibitory effect of ACh (n = 10) could be blocked mostly by M2 receptor antagonist MT (30 nmol/L, n = 7); and this inhibitory effect (n = 9) could be blocked mostly by another M2 receptor antagonist AFDX-116 (30 nmol/L, n = 6), but not by M1 receptor antagonist PZ. PMID- 9863182 TI - [An observation on the possible existance of some information transmission between sensory nerve endings of rat]. AB - Dorsal cutaneous branches of the spinal nerves of T5-L1 segments on one side in anaesthesised rats were exposed and cut centrally. One peripheral end of the cut nerves was selected for antidromical stimulation, while an adjacent one for recording, both by Ag-AgCl electrodes. When a train of 50 Hz, 0.2 ms width square waves of 2 s was delivered to the nerve preparation for stimulation, the discharge of the adjacent nerve showed a remarkable increase and then subsided to the original level for about 25 s. Such experiments (n = 414) showed that the induced discharge frequency reached its maximum 2 s after the stimulation and maintained for another 2 s before subsiding to original level in about a period of 25 s. Using the discharge frequency at a certain time segment as an index, the degree of induced discharges of the recorded nerves were linearly related to their spontaneous discharges before stimulation and the sequences of stimulation to some extent. Single square stimulation was unable to induce one-to-one correspondent discharge in the recording nerve. Control experiments were done to rule out the induced discharges being due to the spreading of stimulating current. To summarize the results of all the above experiments, it appeared that some information transmission between sensory nerve endings may be present through some chemical events. PMID- 9863183 TI - [Characteristics of synaptic responses of motoneurons to ventrolateral funiculus stimulation in vitro]. AB - By using the intracellular recording techniques, the electrophysiological characteristics of fast excitatory postsynaptic potentials (EPSP) evoked by ventrolateral funiculus (VLF) stimulation were analyzed in neonate rat motoneurons (MN) of spinal cord slices. The incidences of VLF-EPSPs was 80% (n = 28), among which 2 is preceded by inhibitory postsynaptic potential and 6 followed by slow EPSPs. Considering the distribution skewness of VLF-EPSP latency histogram, it was suggested that short- and long-latency EPSPs correspond respectively to mono- and poly-synaptic transmission respectively. The possible neurotransmitters mediating VLF-EPSPs were excitatory amino acids and non-NMDA receptors were critically involved in these synaptic transmissions, for both VLF EPSP and glutamate-induced response were similarly and almost completely abolished by kynurenic acid and DNQX. Typical spatial summation of VLF-EPSPs and EPSPs evoked by ventral root stimulation were observed in the same recorded MNs. Most importantly, VLF-EPSPs were found to increase with hyperpolarizing and decreased with depolarizing the membrane, indicating that the descending fibers in the VLF may terminate on the soma or proximal dendrites of MNs, while the EPSPs evoked by the dorsal or ventral root stimulation were insensitive to membrane potential changes, indicating that the primary sensory afferents in the dorsal and ventral roots may contact the distal dendrites of the MNs. The results imply that the commanding signals conducting along the descending fibers in the VLF may directly modulate the MN's activities, whereas messages from the periphery may cause fine changes of the membrane potential subject to integration of the MNs. PMID- 9863184 TI - [Optokinetic nystagmus induced by moving compound gratings]. AB - Optokinetic Nystagmus (OKN) induced by moving compound gratings was investigated. It was found that OKN sometimes tracks the coherent motion of the rigid plaid in a single direction, and sometimes tracks alternately the transparent motion of two component gratings in different directions, as Dual Alternate OKN. It was also found that the duration of OKN tracking coherence increases as the moving speed of the component gratings decreases, and as the angle between their moving directions decreases. The results also showed that the effect of angle plays a dominant role. PMID- 9863185 TI - [The selective suppression of GABA on cone-driven horizontal cells in carp retina]. AB - Intracellular recordings were made from horizontal cells in the superfused, isolated carp retina and effects of GABA on rod- and cone-driven horizontal cells were studied. Bath-applied GABA (0.5-5 mmol/L) resulted in a hyperpolarization of dark membrane potential and a suppression of light responses of L-type cone horizontal cells. In 7 out of 10 cells tested the effects of GABA were completely blocked by bicuculline, an antagonist of GABAA receptors. In the presence of 0.5 mmol/L Co2+, 2 mmol/L GABA could not induce any change in dark membrane potential in 8 out of 12 cells, but cause slight depolarization in other 4 cells. These results suggest that the GABA effects may be mediated by GABAA receptors on cone terminals in the majority of the L-type cone horizontal cells. GABA had no effect on rod horizontal cells. PMID- 9863187 TI - [Phenomenon of intrinsic rhythm of luteinizing hormone release from isolated anterior pituitary gland of female rat]. AB - The intrinsic nature of rthymic release of luteinizing hormone (LH) of isolated human and rat anterior pituitary gland reported independently by Macro Gambacciani and Xie in 1987 can be more directly demonstrated by a computer programme of Time Series-HSY Hidden Periodic Analytic Approach for continuous monitoring the LH output of the perfusate from a perfusion system with in vitro anterior pituitary of SD female rat. The results are as follows: (1) Under various reproductive conditions the average frequency (min/cycle) and amplitude (ng/ml) of the intrinsic rhythm of LH release were quite different: In proestrous group the frequency and amplitude were the highest, being intermediate in the ovariectomized group and lowest in the lactation group. (2) The intrinsic rhythm of LH release could be changed by either peptide or steroid hormones. In proestrous group with 30 min of gonadotropin-releasing hormone (GnRH), stimulation would reduce both frequency and amplitude. In case of lactation, the frequency was unchanged, but amplitude lowered, while in the ovariectomized rat pituitary, the 30 min GnRH stimulation decreased the frequency of release only. The intrinsic rhythm of the LH release could also be influenced by steriod hormones (Ru486 and Anordrin). With 120 min before removal of the anterior pituitary gland the rats receiving i.m. injection of Ru486 (2 mg/kg bw) or Anordrin (2 mg/kg), the results showed that Ru486 decreased frequency, while Anordrin decreased only the frequency to a less extent, both without amplitude affected. (3) Verapamil and EGTA added to the perfusion system did not abolish but only decreased the rhythmic phenomenon by using proestrous pitutary. This suggests that participation of Ca2+ may take place in the intrinsic release of LH. The above results indicated that the intrinsic rhythm of LH release of isolated anterior pituitary gland is different from various reproductive hormonal conditions and capable of being modified by exogenous hormones. The physiological function of the intrinsic rhythm of LH release of anterior pituitary gland remains to be elucidated. PMID- 9863186 TI - [Protection of superoxide dismutase on hypoxia-reoxygenation injury to endothelial cell]. AB - Short term hypoxia induced endothelial cells (ECs) injury, as manifested in increasing lactate dehydrogenase (LDH) release and malondialdehyde (MDA) content, decreasing nitric oxide (NO) production and antioxidant enzyme glutathione peroxidase (GSH-Px) activity and increased intracellular calcium concentration, which were further exaggerated by reoxygenation. Administration of 200 U/ml superoxide dismutase (SOD) before hypoxia could partially prevent EC from such injuries, suggesting that the presence of oxygen free radicals may be one of the main factors involved in hypoxia-reoxygenation injury. The ameliorative effect of SOD in case is obviously due to elimination of oxygen free radicals. PMID- 9863188 TI - [Nestin mRNA expression during the development of mouse central nervous system]. AB - Intermediate filament Nestin is a marker of neural precursor cells. Its expression pattern during mouse brain development was studied in the present investigation. A pair of primers based on the sequence of mouse Nestin were found best to be suit for the reverse transcriptional PCR (RT-PCR) with this pair of Nestin Primer. Using RT-PCR, Nestin transcripts were detected during the development of the central nervous system (CNS) of mouse, from embryonic day 10 to adult brain. We found that the Nestin showed a transient expression in the development of cerebrum. At the embryonic day 14, the expression of Nestin mRNA reached the highest peak, and was then down-regulated. In the postnatal cerebellum, Nestin gene was also expressed in a transient pattern. The highest peak of Nestin mRNA expression emerged at postnatal day 5, and was then down regulated. Nestin mRNA expression in 10 kinds of adult mouse tissues did not yield any positive results. PMID- 9863189 TI - [Effect of norepinephrine on the thermosensitive neurons in preoptic area of hypothalamus tissue slices in cold acclimatized rats]. AB - In this work, single unit firing activities were recorded in the preoptic anterior hypothalamus (POAH) brain slices of cold acclimatized and room temperature housed rats (CR and RR) and the effects of NE on the neuronal discharges were observed. The neurons of POAH in CR became much more sensitive to NE than that in RR (the threshold concentration of NE of CR became significantly lowered). In comparison with RR, the percentage of warm sensitive neurons that could be excited by NE was decreased and some of them even showed inhibitory responses. On the other hand, the percentage of cold sensitive neurons that could be inhibited by NE was decreased and some of them even showed excitatory responses. The percentage of thermo-insensitive neurons that could be either excited or inhibited by NE were increased. PMID- 9863190 TI - [Effects of hypoxia and anisodamine on products of TXA2 and PGI2 in cultured intra-pulmonary arteriolar smooth muscle cells]. AB - The changes of contents of TXB2 and 6-Keto-PGF1a were studied in severely acute hypoxic cultured intra-pulmonary arteriolar smooth muscle cells (PASMCs) under the action of anisodamine. The results demonstrated that the contents of TXB2 and 6-Keto-PGF1a and their ratio were significantly increased in severe acute hypoxic PASMCs' medium. The content of TXB2 decreased significantly, but the content of 6 Keto-PGF1a was hardly affected by anisodamine under normoxia and hypoxia. These findings suggest that acute and severe hypoxia results in pulmonary vascular constriction through increased production of PASMCs and liberation of TXA2, or PGI2, and increased TXA2/PGI2 ratio. The latter effect of hypoxia could be prevented by anisodamine, which antagonized the effect of hypoxia induced pulmonary vasoconstriction. PMID- 9863191 TI - [Roles of intercellar Ca2+ and protein kinase C played in tissue factor pathway inhibitor and tissue factor expression in human umbilic vein endothelial cells]. AB - The purpose of the present study was to elucidate the roles that protein kinase C (PKC) and calcium played in the tissue factor (TF) synthesis and tissue factor pathway inhibitory (TFPI) release in human umbilic vein endothelial cells (HUVEC). A23187 was used to represent calcium ionophore and phorbol 12-myristate 13-acetate (PMA) as that of PKC activator. TF activity in the lysed HUVEC was measured using one stage clotting assay. TFPI activity in the conditioned medium of HUVEC was assessed by the two-step chromogenic method. The results showed that the TF activities in A23187, PMA and A23187 + PMA groups were remarkably higher (P < 0.01) than that in control. Among the three treated groups, the TF activities in both A23187 group and A23187 + PMA group were lower than that in the PMA group (P < 0.05), but the difference between the former two groups was statically insignificant (P > 0.05). In contrast to the control group, the TFPI activity in the A23187 group was not statistically different (P > 0.05). However, the TFPI activities in the PMA group and the A23187 + PMA group were markedly higher than those in the control group and the A23187 group (P < 0.01). These findings indicate that PKC and calcium ion promote TF synthesis in HUVEC but the effect of the former is stronger than that of the latter, and that the release of TFPI from HUVEC is facilitated by PKC and not significantly affected by calcium ion. PMID- 9863192 TI - [Role of spinal alpha adrenoceptor in the inhibition of renal sympathetic nerve activity and natriuresis induced by blood volume expansion in rabbits]. AB - The effect of spinal alpha adrenoceptor blockage on the inhibition of renal sympathetic nerve activity (RSNA) and natriuresis induced by blood volume expansion was investigated in anesthetized and bilateral sinoaortic denervated rabbits. In the groups of rabbits with intrathecal injection of alpha adrenoceptor blocker phentolamine or artificial cerebrospinal fluid the inhibition of RSNA induced by blood volume expansion were (-25.4 +/- 5.4)% and ( 42.5 +/- 5.2)% respectively (P < 0.05). In the groups of rabbits with intrathecal injection of alpha 1 adtenoceptor blocker prazosin or artificial cerebrospinal fluid the inhibition of RSNA induced by blood volume expansion were (-29.3 +/- 6.1)% and (-42.5 +/- 5.2)% respectively (P < 0.05). These results suggested that both spinal alpha and alpha 1 adrenceptor blockage with attenuated the inhibition of RSNA induced by blood volume expansion. The spinal alpha 1 adrenceptor blockage with intrathecal injection of prazosin also attenuated signiticantly the natriuresis and diuresis induced by blood volume expansion (P < 0.05). PMID- 9863193 TI - [The relationship between the activity of phospholipase A2 and acute hypoxic pulmonary arterial pressure]. AB - The action of phospholipase A2 (PLA2) and related inflammatory mediators on the formation of hypoxic pulmonary arterial hypertension was studied. 30 Sprague Dawley rats were equally divided into three groups at random: normal control group, hypoxic group and the group pretreated with dexamethasone plus hypoxia. The pulmonary arterial pressure (PAP) was measured by inserting a microcatheter into the pulmonary artery. After 30 min of hypoxia, the activity of PLA2, platelet activating factor (PAF), prostaglandin E2 (PGE2) and thromboxane B2 (TXB2) were measured in blood and lung tissue, and it was found that the mean pulmonary arterial pressure (mPAP), the PLA2 activity, PGE2, TXB2 and PAF in blood and lung tissue were significantly increased; but pretreatment with dexamethasone relieved the changes mentioned above. In hypoxia, a positive correlations was found between the PLA2 activity and mPAP, PAF, PGE2, TXB2 respectively; positive correlations were also found between PAF, PGE2, TXB2 and mPAP. In conclusion, PLA2 induced the release of inflammation mediators, which may play roles in the formation of the acute hypoxic pulmonary arterial hypertension. PMID- 9863194 TI - [New species of Natronobacterium]. AB - An extreme haloalkalophilic bacterium HAM-2 with pleomorphic rods was isolated from Hamatai soda lake in Inner Mongolia Autonomous Region. Growth occurs in 12% 30% NaCl, no growth below 12%, optimum 17.5%. pH range for good growth 7.8-10.4, optimum 9.0-9.5. The strain HAM-2 is non-motile, Gram-negative, pleomorphic, cell of which were 1.0-2.0 by 2.0-5.0 microns in size in liquid medium. The major polar lipids of the organisms are phosphatidylglycerol phosphate and phosphatidyglycerol and contains an unidentified phospholipid as a minor component PL4. G + C contant of DNA is 59.5 mol%. Based on the characteristics, the strain HAM-2 could be included in the genus Natronobacterium, but since HAM-2 differs from reported four species of this genus in cell shape, polar lipids and biochemical features. The name Natronobacterium innemongoliae sp. nov. is proposed. The type strain is designated HAM-2. PMID- 9863195 TI - [Expression of green fluorescent protein with baculovirus vector in insect cells]. AB - The green fluorescent protein (GFP) gene was subcloned into the transfer vector pVLneo downstream of the polyhedrin gene (ocu) promoter. Insect cells were cotransfected with recombinant plasmid and Autographa californica Nuclear Polyhedrosis Virus (AcNPV) DNA. In the presence of G418, the recombinant virus containing GFP gene was purified. The GFP expressed in insect cells with a Mw of 30 kDa is observable by strong green light under a fluorescent microscope. Excitation and emission spectra of the GFP were 395 nm and 509 nm respectively. Integration of GFP gene on AcNPV genome was identified directly by Southern blot which gave strong hybridization signal between GFP cDNA probe and 1 kb EcoRI fragment of recombinant virus. PMID- 9863196 TI - [Studies on selectivity of recognizing factor for rmf promoter]. AB - The truncted DNA template carrying rmf gene promoter was transcripted by E. coli RNA polymorase holoenzyme (E sigma) reconstituted with core enzyme (alpha 2 beta beta') and sigma 70 or sigma 38 in vitro. The initional site of the transcription of rmf was confirmed with different restriction endonuclease. rmf promoter can be recognized by E sigma 70 but not E sigma 38. The suitable temperature for in vitro transcription was 37 degrees C, NaCl concentration was 50 mmol/L. PMID- 9863197 TI - [Morphogenesis of AcMNPV nucleocapsid]. AB - In this paper we reported that the nucleocapsid morphogenetic process of AcMNPV in which polyhedrin gene was deleted in the nuclei of sf9 cells. First, the capsid proteins entered the nuclei and assembled many 34nm diameter long fascicularly arranged hollow-tube structures. Then viral DNA entered these tubes that turned into solid structure with high electronic density. The solid structure separated at a certain distance to form 34 x 26 nm fascicular bacilliform structure known as nucleocapsid. Finally, fascicular nucleocapsids were wrapped by envelope and became complete multicapsid morphotype viron. PMID- 9863198 TI - [Nucleotide sequence of gltB gene encoding the large subunit of Rhodobacter sphaeroides glutamate synthase]. AB - The complete nucleotide sequence of a 5.4-kb chromosomal EcoRI-SalI fragment was determined, which contains the structural gene (gltB) for the large subunit of Rhodobacter sphaeroides glutamate synthase, as well as the 5'- and 3'- flanking regions. A open reading frame of 4636 base pairs was identified as R. sphaeroides gltB gene. The MW of the large subunit, as deduced from the nucleotide sequence, was estimated as 164kD. Comparision of the nucleotide sequences revealed a high similarity among gltB genes of R. sphaeroides, Azospirillum brasilense and Escherichia coli. The deduced amino acid sequence of R. sphaeroides GltB showed a high similarity with that of A. brasilense GltB. PMID- 9863199 TI - [Purification and some properties of superoxide dismutase from Fusarium moniliform]. AB - Superoxide dismutase from Fusarium moniliform was purified by the steps including heating, ammonium sulfate fractionation, Sephedax G-100 gel filtration and DEAE Sephadex A-50 chromatography. The results showed that the enzyme was a Mn-SOD with the specific activity of 2640 U/mg and had two homogenous subunits whose molecular mass were 14.5 kD. The wave length of max. absorbing peak in ultraviolet spectrum was 276 nm which was not similar with other resource of SOD. The composition of amino acid was also analyzed. PMID- 9863200 TI - [Laser Raman spectral analysis of superhelical spatial conformation on plasmid DNA]. AB - The pBR322 recombinant plasmids DNA products, which purified by phenol-extract and equilibrium centrifugation in CsCl-ethidium bromide gradients, have been shown as mainly spatial conformation of covalently closed circular DNA (i.e. cccDNA) by agarose gel electrophoresis for homogeneity detection. Laser Raman spectra in the region 450-1750 cm-1 have been obtained for the circular double stranded plasmids DNA molecule in an aqueous solution, indicating not only contain those marker peaks of secondary structure as in conventional B-form of DNA, but also present both 854 and 1083 cm-1 diagnostic bands of reflecting the vibration state of deoxyribosyl phosphodiester backbone. The present analysis have demonstrated relationship between the superhelical state for cccDNA and the two conformational marker bands that can be considered as the tertiary structure marker on plasmids DNA. Because of markedly Raman hypochromicity of charecter band 1378cm-1 of dT in contrast to liear DNA molecule, as well as the carbonyl double bond vibration line of dT have shifted to higher wave number position, the base stacking analysis represent occurrence of both increase in staking reaction activity of dT, and injury of a number of the Hoogsteen hydrogen bond between dA and dT with exist supercoil structure in cccDNA. PMID- 9863201 TI - [Polymorphism and pologenetic affinities of genotype in pathogenic yeast]. AB - The classification of pathogenic yeast, which is one of the most common reasons of opportunistic infection in human body, will be useful for the epidemiological survey and clinical works. The genotypic polymorphism of intergenus, interspecies and intraspecies in 48 isolates, including Candida, Cryptococcus, Torulopsis, Trichosporon, Saccaromyces had been administered carefully with patterns amplified by RAPD (Randomly amplified polymorphic DNA) in 53 primers. The pologenetic affinities had been evaluated by similarity coefficients obtained from these profiles. The results indicated that there are significant difference among intergenus, interspecies and intraspecies in Candida and related yeast. The similarity coefficients among Candida and Crytococcus, Trichosporon were maintained about 80%. The similarity coefficients among interspecies of Candida ranged from 82%-87%, and ones of intraspecies of different Candida species were more than 90%. The genotypic typing of species except C. guilliermondii seems to be related to morphological classification. PMID- 9863202 TI - [Study of the relationship between the control of environmental microorganism and infection of patients with leukemia after trasplantation of bone marrow]. AB - After the transplantation of bone marrow, patients with leukemia are easily infected by kinds of microorganism and die. In this paper, F-mice with the 60Co gamma irradiation (maximum lethal dose) were transplanted bone marrow, then control the microorganism in the environment. Morever the laminar flow wards for the leukemia patient of bone marrow transplantation were designed according to the data combined with the clinic condition. There is no any infection in twelve patients with bone marrow transplantation from 1989 to 1993. PMID- 9863203 TI - [Relation of Lac promotor and the expression of cholera toxin subunit B gene in recombinant Escherichia coli MM2]. AB - Effects of different carbon sources including glucose, lactate and acetate and IPTG induction on the expression of ctb gene, which is on the downstream of lac promotor, in recombinant Escherichia coli MM2 were studied. In medium YC were added 0.048mol/L glucose, 0.102mol/L lactate or 0.167mol/L acetate which separately produce the same energy in the condition of complete oxidization. Addition of glucose largely decreased the expression level of ctb gene because of decrease of pH during culture process. Addition of lactate increased the expression level of ctb gene by 1.15 fold and did not inhibit the growth of MM2 strain. Addition of acetate increasd the expression level of ctb gene by 0.97 fold and inhibited the growth of MM2 strain. Induction by IPTG at different time and different concentration did not increase the expression level of ctb gene, so the lac promotor had no or a little influence upon the expression of ctb gene in recombinant MM2 strain. PMID- 9863204 TI - [The identification and isolation for Pseudomonas vesicularis]. AB - A Gram-negative rod bacteria isolated from the hydrocardia of a hydropericarditis patient was reported. Maincharteristics of the strain are respiratory metabolism, oxidse positive and motility by one polar flagella. Through systematic morphological, physiological and biochemical identification; calculation of G+C mol%, and examination with Biolog Automated Systems, the species of Pseudomonas vesicularis are identified. We pointed out that the strain can infect an immuno suppressed individual. The change of taxonomic position of the species was discussed. PMID- 9863205 TI - [Identification and expression of ribulose 1,5-bisphosphate carboxylase/oxygenase gene from Thiobacillus versutus]. AB - The chromosomal DNA of Thiobacillus versutus was hybridized with various heterologous ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) gene as probes. Only Rhodobacter sphaeroides form I RubisCO gene showed homology with T. versutus. The RubisCO gene fragment of T. versutus was isolated using the RubisCO gene of R. sphaeroides as a probe. And the RubisCO gene of T. versutus can express in E. coli cell. PMID- 9863206 TI - [Screening of thermophilic superoxide dismatase producing strain and some properties of the enzyme]. AB - A thermophilic SOD producing strain was obtained from the bacteria preserved in our lab. Its content of SOD was 8774u/g fresh cells. The strain can tolerate 0.4% H2O2 and 70 degrees C, and it has outstanding culture characteristics. It was identified as Bacillus stearothermophilus, called B.S 211-15. Crude SOD was extracted from B.S 211-15, the recovery of total activity was 69.5%, the specific activity was 1793u/mg protein. The enzyme showed fine heat stability, pH stability and good proteinase resistance. The SOD activity didn't decrease after being kept in room temperature for 2 months. The results of inhibition reactions indicated that this enzyme was Fe-SOD. PMID- 9863207 TI - [Study of dermatomycosis and survey of pathogens in troops of Hainan area]. AB - Subtropical area is the prevalent area of dermatomycosis with natural conditions suptable for the growth and proliferation of fungi causing suterficial dermatomycosis. Dermatomycosis not only brings about certain sufferings to the military personnel in peacetime, but also causes nonbattle loss in manpower in war time. In the present work, a survey of dermatomycosis in Hainan subtropical area of China and isolation of the pathogens were carried out. The results were as follows: The morbidity of superficial dermatomycosis was 34.1% and it was manifested clinically as tinea pedis, tinea versicolor, tinea corporis, tinea axillaris, tinea cruris, etc.; The main pathogen causing dermatomycosis in this area was Trichophyton rubrum which accounted for 50.4% of the pathogens isolated and the next was Trichophyton gypseum which accounted for 20.3%; Trichophyton rubrum could cause dermatomycosis of many sites of the body in this area, but the main lesious were tinea corporis and tinea cruris. PMID- 9863208 TI - [The experimental study of Pseudomonas contamination in soft drinks]. AB - Bymeans of this study, we have found out about the situation of Pseudomons contamination in the soft drinks which are manufactured and sold at the area of Nanchang City. In the course of this study, we examined total 416 specimens, in which we found that 69 specimens were positive reaction. The positive rate made up 16.59% of the total specimens. From these 69 positive specimens we separate isolated andidentified the following Pseudomonas 111 strains including 16 different species: P. aeruginosa, 9; P. fluorescens biovars; 11; P. putida biovars, 14; P. syringae pathovars, 5; P. mendocina, 5; P. alcaligenes, 23; P. pseudoalcaligenes, 10; P. cepacia, 17; P. solanacearum, 1; P. testosteroni, 1; P. delafieldii, 3; P. facilis, 6; P. flava 1; P. psenudoflava, 2; P. palleronii, 1; and a new species of Pseudomonas--P. halosensibilis. The achievement of this study will provide scientific basis for working out the standards of food safety control and inspection, raising the level of food hygieneinspection, and directing the drinks menufacturers and selling units to strengthen the food safety control and inspection. PMID- 9863209 TI - [Studies on microbial genome]. PMID- 9863210 TI - [The mutiplication of Sindbis virus and host BHK-21 cell apoptosis]. AB - The process of apoptosis of BHK-21 cells induced by Sindbis virus (SbV) infection is reported here in details. The nuclear DNA cleavage can be first detected at 6 h after SbV infection, followed by the chromatin margination and condensation at 12 h and the DNA ladder can be detected at 24 h after infection. The details of apoptosis body's formation can be revealed by electron microscopy: First, the outer nuclear membrane protrudes where the condensed chromatin accumulates, then the chromatin get into the budding area with the inner nuclear membrane and separate from the nuclear. Progressively, we cloned the gene of SbV nonstructure protein 2 (nsP2) into the eukaryotic expression vector pMAMneo, and it can be expressed transiently. The DNA cleavages, the basic characteristics of apoptosis also can be detected in some cells. Together with the sequence analysis of nsP2 and the other former results, we conclude that the nsP2 may have direct relation with the SbV-induced apoptosis. PMID- 9863211 TI - [Homology analysis of RubisCO gene of Thiobacillus versutus with extremelly acidophilic thiobacilli]. AB - The chromosomal DNA of Thiobacillus thiooxidans and T. versutus was digested with restriction enzymes, blotted to nylon membrane by the way of Southern, and hybridized with the gene probe of ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) from T. ferrooxidans. The result showed that T. thiooxidans exhibited high homology with the probe anc T. versutus was less homology with it. However T. versutus can hybridize with the gene probe of RubisCO from Rhodobacter sphaeroides, which indicated that they had high homology with each other. As RubisCO was highly reserved in the evolutionary, they should be divided into different group. PMID- 9863212 TI - [Screening of Candida albicans fluconazole--resistant mutation strains]. AB - We adopted the method of miminum inhibitary concentration (MIC) assay according to the method recommened by National Committee for Clinical Laboratory Standard (NCCLS), and selected 2 fluconazole--susceptible strains from over 50 strains of Candida albicans which were preserved in our center (these strains were all isolated from clinical patients). The number of the strains are: BMU8945 (MIC value is 0.125 microgram/ml) and BMU8977 (MIC value is 0.25 microgram/ml). In addition, we also picked one strain of ATCC14053 (MIC value is 0.5 microgram/ml) as our research starting strains. After the induction of UV and DES, we obtained nearly 1000 colonies of fluconazole--resistant strains (MIC > 30 micrograms/ml). After subculture, we found that the characterization of drug--resistance of these strains could be inheredited stably. PMID- 9863213 TI - [Study on the inactivated vaccine of Gardnerella vaginalis of fox. VI. Experimental development of the inactivated vaccine of Gardnerella vaginalis of fox]. AB - The aluminum hydroxide gel, propolis and oil adjuvant inactivated vaccines were developed by silecting good immunity, representing principal epidemic serotype I GVF44 of Gardnerella vaginalis of fox in civil. The results showed that the aluminum hydroxide gel vaccine in safety was better than propolis and oil adjuvant vaccines, the most optimal inactivated concentration of formalin is 0.1%. Efficient immunity doses are 4 billon bacteria/1 ml. There were on any bad influences to occur for fox after the aluminum hydroxide gel of three times immunity doses were injected. The inoculated foxes could resist challenge of a hundred ID50 virulent strains after the vaccine was injected to foxes for 21 d. Immunity period of the vaccine is 6 months storage period is 10 months at 4-10 degrees C. The field experiments confirmed that the vaccine possessed accuracy, stability and reliability. PMID- 9863214 TI - [Conditions for the overexpressions of copper--resistant proteins from Escherichia coli]. AB - There were five copper--resistant proteins encoded in the copper resistant plasmid pRJ1004 from Escherichia coli. The conditions for the overexpressions of two of the five, PcoC and PcoE, were reported in this paper. For the overexpression of PcoC using IPTG (Isopropyl-beta-D-thiogalactoside) as inducer, the best conditions were: medium, LB; IPTG concentration, 0.1 mmol/L; inducing time, one and half hours. For the overexpression of PcoE using CuSO4 as inducer, the best conditions were: medium, LB; CuSO4 concentration, 2.0 mmol/L; inducing time, one and halfhours. PMID- 9863215 TI - [Using lyophilization to the preparation of drug paper disks for antimicrobial susceptibility test]. AB - It was found that the paper disks used in the antimicrobial susceptibility test could be prepared by natural drying at 37 degrees C, but the drug distribution between these disks is not so enen as determined by antimicrobial inhibitory test, thus it was difficult to fit the needs of the national standards. Alternatively, we applied the lyophilization technique to the disks preparation and 12 batches, of different antibiotic disks were made out and found that the difference between disks is reduced, and all are meet to the national requirement. It is supposed that the capillary action may play a role in the variation of drug distribution in the natural drying process, and the lyophilization process avoided the capillary action, hence the more even distributed antibiotic disks are produced. The preparation of lyophilized disks is a simple and practical method that could be used in the antimicrobial susceptibility tests in hospital. PMID- 9863217 TI - [Introduction and analysis of the information on projects received and supported in division of microbiology NSFC in 1996]. PMID- 9863216 TI - [Molecular regulation of sigma factor and anti-sigma factor in prokaryote differentiation]. PMID- 9863218 TI - [Analysis of population pharmacokinetics with NONMEM in clinical patients treated with amikacin by intravenous infusion]. AB - Clinical data (n = 275) collected from 52 patients with respiratory tract infection receiving amikacin (AMK) by intravenous infusion were analysed with NONMEM, a computer program designed for estimating population pharmacokinetic parameters. Concentrations of AMK in serum were determined by fluorescence polarization immunoassay (FPIA). A two compartment open model was used for analysing AMK population pharmacokinetics. The influence of body weight (BW), creatinine clearance (CC), administration history (HIS) and state of pathology (chronic obstructional pulmonary disease, COPD) on pharmacokinetics was investigated. The pharmacokinetic parameters of AMK were shown to be influenced by creatinine clearance (CC) and COPD. PMID- 9863219 TI - [Effects of inhibitor of protein kinase C on brain edema formation evoked by experimental cerebral ischemia in gerbils and rats]. AB - Recent studies have suggested that protein kinase C (PKC) may be involved in the formation of brain edema. In this paper, the effects of two kinds of PKC inhibitors, H-7 and matrine, were examined on the brain edema formation in experimental models. The results showed that pretreatment with H-7 6.25 and 12.5 mg.kg-1 prevented the accumulation of water and certain electrolytes in the unilateral hemisphere of the brain evoked by ligation of a single common carotid artery in Mongolian gerbil; pretreatment with matrine 25 and 50 mg.kg-1 reduced the extent of cerebral edema formation evoked by ligation of a single common carotid artery in gerbil and by middle cerebral artery occlusion in Sprague Dawley rats. These results present new evidence for the involvement of PKC in the formation of brain edema. PMID- 9863220 TI - [Effects of enalapril and taurine on left ventricular hypertrophy and arrhythmia in renovascular hypertensive rat]. AB - The effects of enalpril (Ena, 6 mg.kg-1) and taurine (Tau, 30 mg.kg-1) on left ventricular hypertrophy (LVH) and ventricular arrhythmia were studied in two kidney, one clip renovascular hypertensive rats (RHR). From the 9th week after operation, Ena and Tau were given per oral daily for 9 weeks. These drugs significantly decreased the systolic arterial pressure and the weight of the left ventricle. Combination of both drugs was found to reduce the blood pressure further than either drug used alone. Arrhythmias induced by trains of electrical stimuli were more frequent in working hearts isolated from RHR than that from normotensive rats. Ena and Tau could decrease the incidence of this arrhythmias in RHR. The calcium content in the myocardial mitochondria in RHR was increased compared with that in normotensive rats. Treatment with Ena and Tau reduced this increase significantly. These results suggest that chronic therapy with Ena and Tau can induce an attenuation of systemic arterial pressure and reduce the propensity of RHR heart to arrhythmogenesis by limiting cardiac hypertrophy and calcium overload of the myocardium. PMID- 9863221 TI - [Effects of L-malate, an inhibitor of glutamate decarboxylase, on learning and memory in mice]. AB - Oral administration of L-malate 600 mg.kg-1 for 5 days before training can significantly promote memory acquisition, consolidation and retrieval, respectively, on anisodine-, chloramphenicol-, and ethanol-induced memory impairment in mice. Ketamine (8.5 mg.kg-1, i.p.), a selective NMDA receptor antagonist, was found to inhibit the anti-amnesic activity of L-malate. The glutamate (Glu) and GABA content in mice brain was measured by high-performance liquid chromatography with fluorescence detection. The content of GABA was shown to be decreased from 2.3 +/- 0.5 mumol.g-1 to 1.3 +/- 0.5 mumol.g-1 (P < 0.01), so the Glu/GABA ratio was increased significantly. These results suggest that the decrease of cerebral GABA level and the increase of Glu/GABA ratio is favorable to learning and memory. PMID- 9863222 TI - [Pharmacokinetics and disposition of triptonide in rats]. AB - Triptonide, isolated from Tripterygium wilfordii Hook., was found to show significant antiinflammatory, immunosuppression and antitumor activities. A RP HPLC method was applied to determine the plasma concentration of triptonide at different times in rats. Concentration-time curves after i.v., 0.7, 1.4 and 2.8 mg.kg-1 of triptonide were fitted to a two-compartment open model with T1/2 alpha of 0.167-0.195 h and T1/2 beta of 4.95-6.49 h. The area under curves (AUCs) were linearly related to the dosages (gamma = 0.9894). Systematic clearances (CLs) were independent of dosages. Mean residence time (MRT) of the three doses was 3.26-5.14 h by noncompartmental (the statistical moment method) analyses. The tissue distribution of triptonide in rats appeared to be wide throughout the body. The triptonide levels were high in the lung and liver, moderate in the heart, kidney, spleen and muscle and low in the testis, intestine and brain. Data of the urine and bile excretion indicated that only a small percent of unchanged triptonide was excreted. Plasma protein binding of triptonide rate was about 75%. PMID- 9863223 TI - [Inhibitory effects of tachykinin receptor antagonists on leukotriene C4-induced cardiovascular responses in guinea pigs]. AB - This study is to determine whether sensory neuropeptides are involved in the cardiovascular effects of leukotriene C4 (LTC4). LTC4 (0.8 nmol.kg-1, i.v.) caused hypotensive response and increased Evans blue extravasation from the atria and ventricles in anaesthetized guinea pigs. CP-96345 (2.06 mumol.kg-1, i.v.), a tachykinin NK-1 receptor antagonist, and SR-48968 (1.66 mumol.kg-1, i.v.), an NK 2 receptor antagonist, partially inhibited LTC4-induced increase (46.6% and 37.5%, respectively) of dye extravasation from the atria of guinea pigs. Combination of CP-96345 and SR-48968 markedly inhibited LTC4-induced hypotension and increase of microvascular leakage in both atria and ventricles (58.1% and 54.1%, respectively), similar to the inhibition by ONO-1078 (0.06 mumol.kg-1, i.v.), a specific leukotriene antagonist. These results suggest that NK-1 and NK 2 receptors may be involved in the hypotension and the inflammation of heart induced by LTC4. PMID- 9863224 TI - [Isolation and structure elucidation of cirensenosides O and P from the leaves of Oplopanax elatus Nakai]. AB - Two new triterpenoid saponins were isolated from the leaves of Oplopanax elatus Nakai. By measuring physical and chemical constants and spectral data, their structures were elucidated as 3-epi-oleanolic acid 28-O-alpha-L-rhamnopyranosyl (1-->4)-beta-D- glucopyranosyl (1-->6)-beta-D-glucopyranoside; 3-O-beta-D glucopyranosyl betulinic acid 28-O-beta-D-glucopyranosyl (1-->6)-beta-D glucopyranoside and named as cirensenosides O and P, respectively. PMID- 9863225 TI - [Determination of five isoflavones in Belamcanda Chinensis by RP-HPLC]. AB - A reversed phase high performance liquid chromatographic method for the quantitative determination of the five isoflavones (tectoridin, iridin, tectorigenin, irigenin and irisflorentin) in the rhizome of Belamcanda and Iris was developed. The column was packed with TSK gel ODS 80 TS (150 mm x 4.6 mm). Methanol-water (36%-60%) was used as mobile phase. The method was rapid, sensitive, precise and has good reproducibility. The five isoflavones in the crude drugs were completely separated within 40 min. Recoveries of the five constituents were 98.41%-101.93%. The crude drugs were analysed as follows: The powdered sample was refluxed with methanol for 6 hours. The extract was transferred into a 2 ml volumetric flask and diluted to mark with methanol. A definite amount of this sample solution was injected into RP-HPLC. The contents of the five constituents in the rhizomes of Belamcanda and Iris were calculated from the relevant peak heights or peak areas. The analytical results of determination for various species and habitats are shown in Tab 2-5. PMID- 9863226 TI - [Investigation on in vivo metabolism of 6-methoxy n-butyl phthalide]. AB - A GC-MS method for the investigation on the metabolism of 6-methoxy n-butyl phthalide (MBP) is described. After oral administration of MBP, the rat urine sample was collected, hydrolyzed with beta-glucuronidase, extracted and concentrated for TMS derivatization, and then analyzed by GC-MS. MBP and its six oxidative metabolites were determined in the 0-24 h rat urine sample. The mass spectra of the metabolites and their derivatives were presented and the in vivo metabolic pathway was discussed. PMID- 9863227 TI - [(Sp)-8-chloroadenosine 3',5'-cyclophosphate induced differentiation on human leukemia HL-60 cells]. AB - (Sp)-octyl 8-chloroadenosine 3',5'-cyclophosphate(OCC), a newly synthesized cAMP analog, strongly induced growth inhibition and differentiation in human leukemia HL-60 cells. The effects were dose- and time-dependent and irreversible. In flow cytometry, OCC brought about a block at the G1 phase of HL-60 cell cycle. Determined by incorporation assay, OCC was shown to strongly inhibit DNA synthesis without affecting the synthesis of RNA and protein in HL-60 cells. OCC activated the protein kinase A(PKA) in the cytosol of HL-60 cells and inhibited its binding to cAMP. The activities of PKA in the cytosol of HL-60 cells treated with OCC were more significantly increased than those in control cells. It can be concluded that OCC binds itself to PKA in competition with cAMP and, as a result, activates PKA. PMID- 9863228 TI - [Effects of m-Nif and Nif on left ventricle hypertrophy in rats induced by partial ligation of abdominal aorta]. AB - Partial ligation of the abdominal aorta of rats was adopted to induce left ventricle hypertrophy (LVH). The effects of m-nifedipine (m-Nif) and nifedipine (Nif) on prevention of hypertrophy and the possible mechanism were investigated. The wet weights of the left ventricle (WWLV) of the LVH group were increased compared with those of the sham operated group. After treatment with m-Nif and Nif for 4 wks, the WWLV decreased by 25% +/- 9% and 16% +/- 9%, respectively. The pressure-volume (P-V) curve of the hypertrophied group was markedly elevated, which means that the myocardial compliance was decreased, and the stiffness coefficient of the hypertrophied group was significantly elevated than that of the sham operated group, but the groups treated with m-Nif or Nif were significantly improved. The characteristics of the left ventricular myocardial DHP binding sites were studied. The results showed that the Kd and the Bmax were similar in the m-Nif, Nif and hypertrophied groups, but the total number of the DHP receptors (TNR) of the LVH group was markly increased than that of the sham operated group, but the TNR of the m-Nif and Nif groups were the same as that of the sham operated group. These results suggest that the effects of m-Nif and Nif on preventing cardiac hypertrophy and improving myocardial compliance may be related to their depressing the TNR of DHP. PMID- 9863229 TI - [Antagonistic effect of tetrahydroproberberine homologues on alpha 1 adrenoceptor]. AB - The antagonistic effect of tetrahydroproberberine (THP) homologues on alpha 1 adrenoceptor was studied by combination of radioligand binding assays and measurements of vasoconstriction responses. The results showed that l tetrahydropalmatine (l-THP), l-stepholidine (l-SPD), THPB-18 and tetrahydroberberine (THB) competitively inhibited the 125I-BE2254 specific binding in rat cerebral cortex with pK1 values of 5.54 +/- 0.36, 5.56 +/- 0.47, 5.75 +/- 0.56 and 6.01 +/- 0.60, respectively, and the Hill efficiency was not significantly different from unity. They inhibited phenylephrine-induced constrictions with pA2 values of 5.48 +/- 0.58, 5.66 +/- 0.54, 5.64 +/- 0.34 and 5.45 +/- 0.76, respectively, and the slopes of Schild plot were not significantly different from unity. The results indicate that the 4 THP homologues are non subtype selective competitive antagonists for alpha 1-adrenoceptor with similar affinities. PMID- 9863230 TI - [Contragestational effects of dihydroartemisinin and artesunate]. AB - In experiments carried out in mice, hamsters, guinea pigs and rabbits both dihydroartemisinin and artesunate showed contragestational effect. In mice and rabbits they caused embryo absorption whereas in hamsters and guinea pigs they induced abortion. The contragestational ED50 of dihydroartemisinin given sc on d 7 of pregnancy in mice and d 5 of pregnancy in hamsters were 32.8(27.7-38.9) mg.kg-1 and 6.1(5.6-6.7) mg.kg-1 respectively. The ED50 of this drug given im on d 18 of pregnancy in guinea pigs was 18.3(13.9-24.2) mg.kg-1. Dihydroartemisinin also showed mid-pregnancy terminating effect in hamsters. The contragestational ED50 of artesunate given sc on d 5 of pregnancy in hamsters and the ED50 of sodium artesunate given sc on d 5-8 of pregnancy in hamsters were 12.2(10.3-14.4) mg.kg-1 and 1.0(0.9-1.2) mg.kg-1 daily respectively. Results of light microscopic examination revealed that dihydroartemisinin was selectively toxic to embryo sac. At dose levels sufficient to induce embryo sac necrosis, dihydroartemisinin did not injure the uterus and ovary of the maternal animals. On the ground of the foregoing observations we consider that dihydroartemisinin, artesunate and their analogous drugs should not be used to treat malaria in pregnant women and there is the possibility to exploit intentional abortion agents from artemisinin derivatives. PMID- 9863232 TI - Pharmacokinetic-pharmacodynamic analysis of changrolin in dogs with arrhythmia. AB - The pharmacokinetics and pharmacodynamics of changrolin (CRL) were studied in 7 dogs with arrhythmia induced by coronary artery ligature. The ECG and the percentage of reduction ratio of ventricular premature were used to evaluate the effect of CRL, and an HPLC method was used to determine the serum drug concentration. A pharmacokinetic program was used to fit concentration-time (C-T) data and a combined pharmacokinetic-pharmacodynamic model was used to analyze effect-time (E-T) data in individual dogs. After infusion with CRL 83.33 micrograms.kg-1.min-1 for 60 min, it was found that K10, T1/2, Vd, Cl and Ce were 0.0087 min-1, 78.03 min, 40.55 ml.kg-1, 0.42 ml.kg-1.min-1, and 2.01 micrograms.ml-1, respectively. PMID- 9863231 TI - [Effect of matrine on mouse hepatitis and tumor necrosis factor production induced by Propionibacterium acnes/lipopolysaccharides]. AB - The effect of matrine (Mat) on lipopolysaccharides (LPS)-induced fatal hepatitis and tumor necrosis factor (TNF) production in Propionibacterium acnes (PA)-primed mice were studied. Mice were injected i.p. LPS (10 micrograms/mouse) 7 d after i.p. PA (0.5 ml/mouse) to induce fatal hepatitis. After i.p. LPS, serum TNF activity rose to 1657 +/- 406 kU.L-1 at 1.5 h and ALT activity increased up to 1,496 +/- 890 U.L-1 at 5 h. Six of 8 mice died within 5 h and the massive hemorrhagic necrosis of the liver was observed in all mice. Administration of Mat (10, 50 mg.kg-1, i.p., bid x 3 d) before the LPS injection markedly reduced the elevation of serum TNF and ALT activity in a dose-dependent manner, and diminished the mortality induced by LPS. Liver congestion and necrosis induced by LPS in PA-primed mice were ameliorated markedly by Mat pretreatment. Mat (62.5 250 mg.L-1) inhibited LPS-induced TNF release from PA-primed mouse peritoneal macrophage in vitro in a concentration-dependent manner. These results seggest that Mat protected PA-primed mice from the development of fatal hepatitis induced by LPS due to inhibition of TNF production. PMID- 9863233 TI - [Structure of gypentonoside A from Gynostemma pentaphylum Makino]. AB - A new saponin named gypentonoside has been isolated from Gynostemma pentaphylum (Thunb) Makino, C54H88O21, mp 272-274 degrees C. Its structure was established on the basis of chemical and spectroscopic (IR, FAB-MS, UV, 1H-NMR, 13C-NMR, 2D-NMR) analyses. PMID- 9863234 TI - [Studies on the chemical constituents of seed from Nigella glandulifera]. AB - Seven compounds were isolated from the seed of Nigella glandulifera. Their structures were identified as kaempferol-3-O-beta-D-galactopyranosyl (1-->3)-beta D-glucopyranosyl(1-->3)-beta-D-glucopyranoside (N-I), 2-O-[alpha-D galactopyranosyl (1-->4)-beta-D-glucopyranosyl]-beta-D-fructofuranoside (N-II), N, N-dimethyl-1, 2-dimethoxy-10, 11-dihydric aporphine quaternary ammonium chloride (N-III), 3-O-[beta-D-xylo-pyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1- >2)-alpha-L-arabinopyranosyl]- 28-O -[alpha-L-rhamnopyranosyl (1-->4)-beta-D glucopyranosyl (1-->6) beta-D-glucopyranosyl]-hederagenin (N-IV), sucrose(N-V), beta-sitosterol(N-VI) and cyclolandenol(N-VII). Compounds N-I and N-II are new compounds, named nigeglanoside and nigeglanose, respectively. Apart from ten fatty acids in its oil have also been analysed. It is the first time for the study on chemical constituents of the seed of Nigella glandulifera. PMID- 9863235 TI - [Electroanalytical characteristics of ofloxacin in micellar solution]. AB - In a pH 6.30 buffer solution containing 0.001% Tween-80, ofloxacin (OFX) gives a sensitive polarographic wave at -1.46 V (vs SCE), which can be used for the determination of OFX down to 10(-8) mol.L-1. The linear range is from 1.39 x 10( 7) to 1.39 x 10(-5) mol.L-1. The proposed method was applied to determination of OFX in urine and serum samples with relative standard deviation less than 7.0%. PMID- 9863236 TI - [Studies on oral sustained-release microspheres of nifedipine]. AB - Nifedipine has been incorporated into Eudragit RL microspheres by the emulsion solvent-evaporation process using an acetone/liquid paraffin system. The drug in the Eudragit RL microspheres was present in either molecular dispersion or amorphous state. The drug release profile from microspheres conformed to the Higuchi equation. The microspheres were stable under different storage conditions for six months with respect to drug content, dispersing state and release profile. The in vivo results in eight healthy volunteers showed that the relative bioavailability of the nifedipine sustained-release microspheres against commercial retard tablet as a control was 102.5%, and the microsphere formulation was bioequivalent to the retard tablet. Significant in vitro and in vivo correlation was also found for the microspheres. PMID- 9863237 TI - [Retardation of human drug-resistant HL-60 cell in G1 phase and induction of sensitive cell to apoptosis by cyclosporine A]. AB - To further study the relationship between resistance to apoptosis and drug resistance in harringtonine-resistant HL-60 cells (HR20), cyclosporine A (CsA) 20, 10 micrograms.ml-1 was shown to induce the sensitive HL-60 cells to apoptosis, showing a typical DNA "ladder" band. But the same concentrations of CsA retarded the HR20 cells in G1 phase and could not induce the cells to apoptosis. The cellular daunorubicin accumulation increased when HR20 cells were treated with low concentration of CsA and the reversal of drug resistance by CsA was unrelated to the retardation of cell cycle progression. High phosphorylation of about 50 kDa protein occured when HR20 cells were treated with CsA 10 micrograms.ml-1. The results domonstrate that cyclosporine A retarded the harringtonine-resistant HL-60 cells in G1 phase but induced HL-60 cells to apoptosis, and the retardation was unrelated to drug resistance. PMID- 9863238 TI - [The effect of batroxobin on coronary segmental resistance and coronary blood flow in acute myocardial ischemic dogs]. AB - To study the effects of batroxobin on coronary circulation and cardiac performance in acute myocardial ischemia, Batroxobin was given intravenously to dogs with experimental coronary stenosis. A dose-dependent increase of coronary blood flow (CBF) was observed. Forty minutes after batroxobin (2 BU.kg-1 at infusion rate 0.1 BU.kg-1.min-1) administration, CBF increased by 12% (P < 0.05), small coronary resistance(RS) decreased from 4.1 +/- 0.5 to 3.2 +/- 0.5 mmHg.min.ml-1 (P < 0.01), while large coronary resistance(RL) changed insignificantly from 3.9 +/- 0.8 to 3.8 +/- 0.7 mmHg.min.ml-1 (P > 0.05). Two hours following drug administration, the changes in CBF, RS and RL still remained and RT decreased by 13% (P < 0.05). The + LV(dp/dt)max and -LV(dp/dt)max increased by 14% and 16% (P < 0.05) respectively compared with those in control group. It is concluded that batroxobin improves the ischemic canine coronary circulation and cardiac performance by way of lowering the small coronary resistance and thus increasing CBF. The data also suggest the benificial effect of batroxobin in acute myocardial ischemia. PMID- 9863239 TI - [Inhibitory effects of m-nifedipine on angiotensin II stimulated proliferation and protein synthesis in cultured rabbit aortic smooth muscle cells]. AB - By using rabbit aortic smooth muscle cells (VSMC) in culture as a model, the effects of m-nifedipine (m-Nif) on the growth of VSMC were observed. The results showed that m-Nif inhibited [3H]-thymidine and [3H]-leucine incorporation into DNA and protein, decreased cell number in a concentration-dependent manner in angiotensin II (ANG II) induced VSMC. At the concentration of 2 mumol.L-1, m-Nif was found to reduce the ANG II (100 nmol.L-1) stimulated [3H]-thymidine, [3H] leucine incorporation and cell number by 46%, 58% and 53%, respectively. Our results suggest that m-Nif can inhibit ANG II stimulated proliferation and protein synthesis in cultured rabbit aortic VSMC. PMID- 9863240 TI - [Two-site absorption model fits to pharmacokinetic data of gemfibrozil in man]. AB - The plasma concentration-time data of gemfibrozil in 8 male subjects were determined after an oral dose of 600 mg. Two-peak concentrations in plasma were observed. A kind of one-compartment model with two-sites of drug absorption was proposed and used to fit these data. A good agreement between observed and predicted data was found in all subjects with correlation indexes (gamma 2) > 0.99. The corresponding pharmacokinetic parameters were estimated as follows: Tmax1, 1.10 +/- 0.46 h; Tmax2, 2.60 +/- 0.73 h; Cmax1, 13.62 +/- 4.30 micrograms.ml-1; Cmax2, 17.22 +/- 3.83 micrograms.ml-1; T1, 0.06 +/- 0.06 h; T2, 1.42 +/- 0.57 h and T3, 1.79 +/- 0.60 h. PMID- 9863241 TI - [Differentiation of B16 melanoma cells induced by ginsenoside RH2]. AB - The effect of ginsenoside Rh2, a constituent isolated from Panax ginseng C. A. Meyer, on the growth of tumor cells in vitro was investigated. The results showed that Rh2 inhibited the growth of B16 cells at the concentration of 10 micrograms.ml-1 (IC50: 4.1 micrograms.ml-1). Rh2 was found to significantly induce the activity of differentiation of B16 cells at the concentration of 10 micrograms.ml-1 in vitro. The melanin synthesis of Rh2 in treated B16 cells was increased. Morphologically, the Rh2 induced B16 cells turned to be epithelioid cells. B16 cells became dendrite shaped morphologically at higher concentration of Rh2. Flow cytometry demonstrated that the B16 cells treated with Rh2 were blocked at G1 phase. PMID- 9863242 TI - [Protection of dl-(15R)-15 methyl-PGE2 methyl ester (PG6E) on experimental ulcers in rats]. AB - The effects of dl-(15R)-15 methyl-PGE2 methyl ester (PG6E), in experimental ulcers induced by absolute alcohol, HCl, indomethacin, pyloric ligation and chronic acetic acid in rats were studied. PG6E at doses 10-80 micrograms.kg-1 was shown to have significant protective effect. PG6E (30-60 micrograms.kg-1) was also found to reduce markedly the acid secretion, pepsin activity, DNA content of the juice collected from pylorus ligated stomach of rats and increase markedly the content of gastric mucosa hexosamine in mice given PG6E (30-60 micrograms.kg 1 po for 3 d). At doses of 40-80 micrograms.kg-1, PG6E was able to have no significant effect on gastric emptying time in rats and gastrointestinal tract movement in mice. It appears that PG6E was shown to inhibit the aggressive factors and increase the protective factors of gastric mucosa. This may hopefully become a new antiulcer agent. PMID- 9863243 TI - [Studies on synthetic peptide. XX: the antigenicity and linear epitope map of synthetic peptide hepatitis C virus]. AB - Hepatitis C virus (HCV), the major causative agent of post transfusion non-A, non B hepatitis (NANB), had been cloned and expressed. According to the protein sequence of HCV-BK and its epitope profiles which combined the hydrophilicity, accessibility, flexibility, antigenicity, charge distribution and HPLC reserve coefficient of protein using the "Goldkey" computer program, we designed and synthesized the following peptides: P1(475-495), P3(449-468), P4(658-663), P5(645 663), P6(484-489), P7(475-489), P15(655-662), P16(230-237), P17(225-237), P18(1220-1240), P19(1694-1735), P24(1230-1240), P25(1482-1493), P26(384-389), P27(2355-2389). The results of ELISA showed that P6(60% positive results) and P19(63% positive results) testing with PT-HC of Gu An, Hebei province were the major antigens in NS1 and in NS4 region, respectively. PMID- 9863244 TI - [Studies on the structure of isoastilbin]. AB - A new compound was isolated from Smilax glabra Roxb., named isoastilbin. It was identified as 5, 7, 3', 5'-tetrahydroxyl-flavanonol-3-O-alpha-L-rhamnopyranoside by means of chemical and spectrometric analysis (UV, IR, 1H-NMR, 13C-NMR, 2DNMR and FAB-MS). PMID- 9863245 TI - [New diterpenoids, isodoternifolins A and B, from Isodon ternifolius]. AB - Two new diterpenoids, named isodoternifolin A and B, together with seven known compounds were isolated from the ethanol extract of dried stems and leaves of Isodon ternifolius (D. Don) Kudo. Their structures were determined as 7 beta hydroxy-6 beta, 11 alpha, 15 beta-triacetoxy-7 alpha, 20-epoxyentkaur-16-ene (1) and 6 beta, 7 beta, 15 beta-trihydroxy-11 alpha-acetoxy-7 alpha, 20-epoxy-entkaur 16-ene (2) by chemical and spectral evidence. PMID- 9863247 TI - [Investigation on in vivo metabolism of n-butyl phthalide]. AB - A GC-MS method for the investigation on the metabolism of n-butyl phthalide (NBP) is described. After oral administration of NBP to rats, urine was collected, hydrolyzed with beta-glucuronidase, extracted and concentrated for TMS derivatization, and then analyzed by GC-MS. HBP and its four oxidative metabolites were determined in 0-24 h, 24-48 h rat urine. The mass spectra of the metabolites and their derivatives were presented and the in vivo metabolic pathway was discussed. PMID- 9863246 TI - [Studies on transdermal delivery system of dihydroetorphine hydrochloride]. AB - A transdermal delivery system of dihydroetorphine hydrochloride (DHE-TDS) was developed. The DHE-TDS mainly composed of polyvinyl alcohol, polyvinyl pyrrolidone and lactose. Tests on rabbits showed only slight skin irritation according to federal hazardous substances act. By giving DHE-TDS to rabbits, DHE release was shown to be governed by first-order mechanism. When DHE-TDS was given to Wistar rats, a relatively stable blood drug concentration was observed from 4 32 h after drug administration. Writhing tests showed that one dose of DHE-TDS would maintain the narcotic action on rats for at least 48 h. PMID- 9863248 TI - [Studies on the chemical constituents of Rhodiola fastigita]. AB - The root and rhizoma of Rhodiola fastigita S. H. Fu. is a traditional Tibetan medicine used for promoting blood circulation and relieving cough. In recent years, it was generally used as a tonic. So, its chemical constituents were studied. A new flavonoid and six known compounds were obtained. The known compounds were: 4'-methoxyl herbacetin (I), rhodiolin (II), dihydrokaempferol (III), daucosterol (IV), tyrosyl (V) and salidroside (VI). The structure of the new flavonoid was elucidated as herbacetin-8-O-alpha-D-lyxopyranoside (VII), by means of UV, IR, MS, 1H and 13C-NMR spectral data and chemical methods. PMID- 9863249 TI - [Improvement of learning and memory functions by GABAB receptor antagonists in mice]. AB - In one trial passive avoidance response in mice, the effects of gamma aminobutyric acid (GABA)B receptor agonist baclofen and antagonist CGP35348 and CGP36742 on acquisition, consolidation and retrieval of memory were observed. The results showed that the antagonists could significantly promote the acquisition impairment induced by baclofen, the consolidation impairment induced by baclofen and NaNO2, and the retrieval impairment induced by baclofen and 30% alcohol. These results suggest that the GABAB receptor antagonists may become a novel type of drug for the treatment of Alzheimer's disease. PMID- 9863250 TI - [Effect of norepinephrinergic system on ipalbidine analgesia]. AB - Ipalbidine (Ipa) is a photoactive alkaloid isolated from the seeds of ipomoea hardwickki Hemsl. The analgesic effects of Ipa were determined by rat tail flick method. A dose-dependent analgesic effect was found after s.c. or i.c.v. administration of Ipa, but no analgesia was observed after intrathecal injection, indicating that the analgesic effect of Ipa is central in origin, and it acts mainly on supraspinal substrate. The analgesic effect induced by Ipa (60 mg.kg-1, s.c.) was markedly reduced by reserpine (2 mg.kg-1, i.p.) given 24 h before Ipa, which was reversed by combined administration with i.c.v. norepinephrine (NE). In addition, Ipa-induced analgesia was significantly attenuated by electrolytic lesion of bilateral destruction of locus coeruleus, and combined administration with diethyldithiocarbamate (200 mg.kg-1, i.p.), phentolamine (10 mg.kg-1, i.p. or i.c.v.), and prazosin (3 mg.kg-1, s.c.). But no influence was observed on the analgesia of Ipa after administration of yohimbine (5 mg.kg-1, s.c.) or propranolol (10 mg.kg-1, i.p.). These results suggest that the analgesia caused by Ipa is closely related to the function of the central norepinephrinergic system, and probably mediated by indirectly acting on alpha 1 receptors, but not alpha 2 or beta receptors. PMID- 9863251 TI - [Ketamine and nifedipine protect cultured cortical neurons against injurious effect of glutamate]. AB - The effects of glutamate on cultured cortical neurons and the protective effect of ketamine and nifedipine were studied. On day 10 after plating of the cortical cells from 16-18 day-old fetal rats, the cultures were exposed to 50 mumol.L-1 glutamate and low glucose (1 g.L-1) for 10 min-24 h. The results showed that a release of lactate dehydrogenase (LDH) into the culture supernatant was observed as a function of time. The values of LDH efflux in culture medium was significantly lower than those of controls when the cells were pretreated with ketamine or nifedipine 10 min prior to addition of glutamate. More significant decrease of LDH activity in culture medium was observed when the two drugs were used in combination. These results demonstrate that the dissociated cultured cortical neurons from fetal rat are seriously damaged by glutamate. Such damage could be attenuated by ketamine and nifedipine, suggesting that ketamine and nifedipine may protect neurons from the glutamate toxicity, and the effect of combining ketamine and nifedipine was greater than either ketamine or nifedipine alone. PMID- 9863252 TI - Synthesis and antiprotozoal activities of some new triazine derivatives including a new antitrypanosomal agent: SIPI-1029. AB - Two series of compounds, 1,2-dihydro-2,2-dimethyl-4, 6-diamino-1-(omega haloalkyloxy)-s-triazines and O, O'-bis (4, 6-diamino-1, 2-dihydro-2, 2 disubstituted-s-triazin-l-yl) alkanediols were synthesized and tested against Plasmodium berghei and Trypanosoma evansi in mice. Most title compounds showed good antimalarial activity and compounds IIc-e showed good antitrypanosomal effect. After further studies on pharmacology, toxicology, pharmacokinetics and efficacy on infected cattles compound IIe (SIPI-1029, T-46) was shown to be a new highly active antitrypanosomal agent with low toxicity and long half life in plasma. PMID- 9863253 TI - [Synthesis of optically active (R)- and (S)-tai-ding-an(TDA) and their anti-HSV activity evaluation]. AB - Tai-Ding-An (3-phthalimido-2-oxo-n-butyraldehyde bisthiosemicarbazone, TDA) is an antiviral drug first synthesized in this institute. In order to clarify the difference between the two enantiomeric isomers of TDA, (R)- and (S)-TDA were synthesized from (R)- and (S)-alanine, respectively, via the following steps: fusing with phthalic anhydride gave 2-phthalimido alanine(2a or 2b). The resulting acid was reacted with thionyl chloride to offer the corresponding acid chloride(3a or 3b), which was treated with diazomethane to give the diazoketone(4a or 4b). Bromination of the ketone with hydrobromic acid gave the key intermediate 3-phthalimido-2-oxo-1-bromobutanone (5a or 5b). Compound 5a or 5b was oxidized with DMSO to give 6a or 6b, which was directly condensed with thiosemicarbazide to afford the desired (R)- or (S)-TDA. (R)-TDA, (S)-TDA and (RS)-TDA have been tested in cell culture for anti-Herpes simplex virus I (HSV-1) and HSV-2 activities by plaque reducing method. All of them showed inhibitory effects on HSV-1 and HSV-2 replication with IC50 of 0.0296 mmol.L-1, 0.0359 mmol.L-1 and 0.0418 mmol.L-1 for HSV-1 and 0.88 mmol.L-1, 1.04 mmol.L-1 and 1.06 mmol.L-1 for HSV-2. Not much difference was found among these compounds either on IC50 or on therapeutic indexes. PMID- 9863254 TI - [Synthesis of adenine derivatives and their activities against herpes virus in vitro]. AB - A series of 9-(N4-substituted acetaldehyde thiosemicarbazone) adenines were synthesized and evaluated for antiherpes virus activity. Compounds 4a-l were prepared by condensation of 9-(acetaldehyde) adenine(6) and the corresponding N4 substituted thiosemicarbazides (10). The antiviral effects of all compounds 4a-l were tested in vitro in primary rabbit kidney cell cultures infected with herpes simplex virus type 1 (HSV-1) and varicella-herpes zoster virus (VZV), and in primary human embryo cell cultures infected with herpes simplex virus type 2 (HSV 2). The results showed that the minimum inhibitory concentrations (MIC) of 4e and 4f for HSV-1 and VZV were 20, 40, 20 and 20 micrograms.ml-1, respectively, and other compounds were 200 micrograms.ml-1. For HSV-2, the MIC of all tested compounds were 300 micrograms.ml-1. We also evaluated the antiherpetic effect of 4e (and 4f) by combination with acyclovir (ACV) in the ratio of 1:1 in vitro. The MIC of the combined compounds were 2 micrograms.ml-1 for 4e and 6 micrograms.ml-1 for 4f, while their minimum cytotoxicities (MCC) in the cell were markedly reduced compared with the individual compounds. PMID- 9863255 TI - [New ellagic glycosides and known triterpenoids from Duchesnea indica Focke]. AB - Two new ellagic glycosides, named ducheside A and ducheside B, have been isolated from Ducheside indica Focke. Their structures were established as 3'-O-methyl ellagic acid-4-O-beta-D-xylopyranoside (I), 3'-O-methyl-ellagic acid-4-O-alpha-L arabinofuranoside (II), respectively, on the basis of chemical method and spectral analysis. In addition, four known compounds were isolated and identified as 3 beta-hydroxylurs-12-en-28-oic acid (III), 2 alpha, 3 beta, 19 alpha trihydroxylurs-12-en-28-oic acid (IV), 2 alpha, 3 beta, 19 alpha-trihydroxylurs 12-en-28-oic acid 28-O-beta-D-glucopyranoside (V), 2 alpha, 3 alpha, 19 alpha trihydroxylurs-12-en-28-oic acid 28-O-beta-D-glucopyranoside (VI). Compounds IV, V, VI were isolated for the first time from Duchesnea genus. PMID- 9863256 TI - [Separation and identification of the flavonoids from Buddleia officinalis Maxim]. AB - The flowers of Buddleia officinalis Maxim have been used to cure eye inflammation in China. Eight flavonoid compounds were isolated from them. Their structures were characterized as acacetin (1), apigenin (2), luteolin (3), neobudofficide (4), linarin (acaciin 5), luteolin-7-O-rutinoside (6), luteolin-7-O-glucoside (7) and cosmosiin (8), on the basis of chemical and spectral evidences. The new compound 4 was identified as 5,7-dihydroxy-4'-O-methoxyflavone-7-O-alpha-L rhamnopyranosyl (1-->2)-[alpha-L-rhamnopyranosyl (1-->6)]-beta-D-glucopyranoside and named neobudofficide. All the compounds were isolated from B. officinalis for the first time, except linarin and acacetin. PMID- 9863257 TI - [Study on major metabolites of 3H-1,2-dihydro-2-(4-methylphenylamino)-methyl-1 pyrrolizinone in rabbits]. AB - The metabolites of a 750 mg oral dose of Z-47 [3H-1, 2-dihydro-2-(4 methylphenylamino) methyl-1-pyrrolizinone], a new anti-inflammatory and analgesic agent, in rabbit urine were separated and detected with high performance liquid chromatographic method. On basis of the chromatographic behavior of Z-47 metabolites and biotransformation pathways of drugs with partial structure of Z 47, the carboxylic derivative of Z-47 [4-(3H-1, 2-dihydro-1-pyrrolizinone-2 methylamino) benzoic acid] was proposed as a potential metabolite so that the compound was synthesized. The authentic substance was then compared with one of the metabolites by the chromatographic retention time and the ratio of their UV absorbances at two wavelengths. The enzyme-hydrolyzed product of another metabolite was also analysed. It was consequently confirmed that the carboxylic derivative of Z-47 and its acyl beta-D-glucuronide are major metabolits of Z-47 in rabbits. PMID- 9863259 TI - [Effects of 1-(2,6-dimethylphenoxy)-2-(3,4-dimethoxy phenylethylamino) propane hydrochloride on cytosolic free calcium concentration of rabbit platelets]. AB - The effects of 1-(2, 6-dimethylphenoxy)-2- (3,4-dimethoxyphenylethylamino) propane hydrochloride (DDPH) and verapamil on cytosolic free calcium concentration of rabbit platelets were investigated. The results showed that the elevation of cytosolic free calcium concentration ([Ca2+]i) of rabbit platelets following ionophore A23187 was significantly inhibited by DDPH and verapamil. The rates of inhibition with DDPH and verapamil were 58% and 78%, respectively, with no significant difference between the two groups (P > 0.05). PMID- 9863258 TI - [Blocking effects of benzyltetrahydropalmatine on delayed rectified K+ currents expressed in Xenopus oocytes and in toad oocytes]. AB - The effects of benzyltetrahydropalmatine (BTHP) on delayed rectified K+ currents (Ik) expressed in Xenopus oocytes and Ik of toad (Bufo bufo gargarizans) oocytes were studied. The Ik expressed in Xenopus oocytes was measured after microinjection of mRNA isolated from carp fish (C anratus L.) brains with double microelectrode voltage clamp technique. The maximum and mean value of Ik expressed in Xenopus oocytes were 600 nA and 360 +/- 104 nA, respectively. BTHP reduced the current amplitude of Ik expressed in Xenopus oocytes in 10-1000 mumol.L-1 dose-dependently, EC50 was 29 mumol.L-1. Also, the reduction of Ik of toad oocytes was 9.1%, 29.1%, 54.7% and 68.6% by BTHP 10, 30, 100 and 1000 mumol.L-1, respectively, EC50 was 33 mumol.L-1. The results showed that BTHP possesses an inhibitory effect on Ik, the main ion mechanism of antiarrhythmic action of BTHP. PMID- 9863260 TI - [Studies on the characteristics of leukotriene B4 receptor with radio-ligand binding assay]. AB - Leukotriene B4 (LTB4), one of the metabolites of arachidonic acid via 5 lipoxygenase (5-LO), plays important role in some inflammatory diseases as one of the most potent chemotaxis factor. A radio-ligand binding assay was set up and the characteristics of LTB4 receptor on guinea-pig splenocytes membrane were studied. At 25 degrees C, the Kd was found to be 1.55 x 10(-9) mol.L-1 and the Bmax was 2.59 x 10(-13) mol.mg-1 protein. The assay established was evaluated by nordihydroguaiaretic acid (NDGA) as positive control. PMID- 9863261 TI - [Structure-activity relationship studies of chalcones as SRS-A receptor antagonists]. AB - A simple, reliable and highly sensitive bioassay with sensitized longitudinal strips of guinea pig ileum was used for screening the receptor antagonists of slow reacting substance of anaphylaxis (SRS-A). The SRS-A receptor antagonistic activities of 17 chalcones were studied. Most compounds in these chalcones were found to have SRS-A receptor antagonistic action at the concentration of 10(-4) mol.L-1. Among them, compounds 5, 13 and 17 were highly effective with IC50s of 7.5 x 10(-6), 7.5 x 10(-6) and 6.8 x 10(-5) mol.L-1, respectively. Under the same conditions, the IC50 of FPL 55712, a known leukotriene D4 receptor antagonist, was shown to be 3 x 10(-4) mol.L-1. It would appear that compounds 5, 13 and 17 were 40, 40 and 4.4 times more potent, respectively, than FPL 55712. From analysis of structure-activity relationship of chalcones, these results suggest that the following factors may be important for an active antagonist of SRS-A receptors: (a) There is a system of pi, pi conjugation in the molecule; (b) The ester group in the B ring of chalcones is more favorable than the carboxyl group; (c) Antagonism for meta- or para-substituted derivatives of carboxyl or ester group in the B ring are more potent than ortho-substituted compounds; (d) The length of carbon chain of alkyl group in the A ring of chalcones is more effective for 1, 4 or 6 carbon atoms than for 10 or 14 carbon atoms. PMID- 9863262 TI - Genetics in ophthalmology. PMID- 9863263 TI - [Present status of the iodine supply and its results in the Czech Republic]. AB - Iodine is essential for the synthesis of thyroid hormones. Its deficiency has an adverse impact on thyroid function and leads in particular in early developmental stages to serious irreparable disorders. Our country has been for times immemorial in an area of inadequate iodine supplies, in the past even endemic cretenism was encountered in our country. Other sequelae of iodopenia are goitre, hypothyroidism, impaired somatic and mental development, reduced fertility, increased neonatal and infant mortality and a generally reduced physical and mental quality of the population. Supplementation of table salt with iodine led to a marked improvement of the position, however, recently an increase of goitre was again recorded. This is why pilot studies of iodine supplies and the thyroid state are under way in the Czech Republic. Random population samples from six regions were examined, a total of some 3300 subjects aged 6-65 years. It was revealed that the iodine supplies (evaluated from the iodinuria) is liminal and that 10% of the children and 20% of the adults (more girls and women than boys and men) suffer from severe iodine deficiency (iodinuria 50 micrograms/l in the morning urine). The sonographically assessed volume of the thyroid gland in our country is greater than in areas with adequate iodine supplies. The mean values of thyroid hormones, TSH and the Achilles tendon reflex are within the normal range. Solution of iodine deficiency is one of the fundamental parts of the programme of the UN, WHO and UNICEF. In our country we must, with this objective in mind, improve the iodization of table salt, increase the iodine content of salt and ensure an adequate intake of iodine in pregnant and lactating women by supplementation with iodine tables. An increase of the iodine intake of infants is also essential, in bottlefed infants supplementation of formulas is necessary. The problem cannot be solved without ensuring the use of iodized table salt in the food industry and without ensuring an overall iodine supply from multiple sources. This complex problem calls for an interdisciplinary approach. Only in this way it will be possible to fulfil the pretentious task of elimination of iodine deficiency diseases in children and adolescents by the year 2000. PMID- 9863264 TI - [Chlamydia trachomatis--a risk for reproductive capacity in women]. AB - BACKGROUND: Chlamydia trachomatis is a bacterium which causes in man among others urogenital infections. So far no data were published pertaining to the herd immunity of the population to this infection in the Czech Republic. The objective of the present investigation was to summarize clinical and laboratory results of examined women and to evaluate the assembled data by statistical methods. Special attention was paid to patients examined on account of infertility. METHODS AND RESULTS: 506 women aged 17-50 years (mean age 28 years) were examined. They were divided into five groups (1-cervicitis, 2-pelvic pain, 3-pregnant, 4-sterile, 5 before UPT). Group 4 (sterile) was further subdivided into three categories (4a IVF on account of tubal sterility, 4b IVF on account of non-tubal sterility and 4c others). With regard to the domicile the women were divided into those residing in Prague and those from other towns and rural areas. In all antigenic examinations of smears from the endocervix were made and serological examinations of specific IgA, IgG antichlamydial antibodies. During collection of the smears the appearance of the cervix; discharge, bleeding and pain on examination were evaluated. For statistical evaluation the chi 2 test was used. In 57% of the women from the whole group positive IgG antibodies were detected suggesting a past or present infection. Differences of antigenic or serological positivity between the five different groups were not statistically significant. In category 4a a significantly higher positivity of IgG (2 alpha < 0.001) of antichlamydial antibodies was found (92% patients) than in categories 4b and 4c. 71% women had complaints during examination. The authors did not find a correlation between antigen- and serum positive women indifferent groups and their domicile. CONCLUSIONS: The results provide evidence of a high herd immunity of the population of the Czech Republic to Chlamyata infection. At the same time they prove that post-inflammatory changes caused by Chlamydia trachomatis are the most frequent cause of occlusion of the oviducts and tubal sterility. PMID- 9863265 TI - [Evaluation of diet in a sample of Czech mothers six months after delivery]. AB - BACKGROUND: Health care of nursing mothers and their infants is an important priority of primary preventive care. The mother's diet plays an important role in this respect. The objective of the presented investigation was to assess the adequacy of the dietary intake of lactating mothers during the sixth month after delivery. METHODS AND RESULTS: Data on the education, body weight, height of the mother, dietary intake, evaluated from a three-day dietary record, were collected from 131 nursing mothers and compared with the Czech recommended dietary allowances for nursing mothers as well as with data from 265 controls, i.e. women who did no longer breastfeed their babies. From the results ensues a significantly higher calcium intake (937 mg; SD = 415, p < 0.001), vitamin B1 (1.1 mg, SD = 0.5, p < 0.001), total energy (8.7 MJ, SD = 2.6, p < 0.01), protein 75.4 g, SD = 18, p < 0.01), carbohydrates (281 g, SD = 112, p < 0.01) and riboflavin (1.3 mg, SD = 0.5, p < 0.01) in nursing women as compared with those not nursing. The nursing mothers, however, do not meet the Czech recommended dietary allowances as regards total energy, calcium, vitamin C, linoleic acid, vegetable proteins and iron. Other problematic nutrients-magnesium, zinc, folates, pyridoxine, selenium and iodine could not be assessed as they are not listed in the Czech food composition tables. In women with university education the energy and nutrient intake was in the majority more favourable than in women with elementary education. On the other hand no statistical differences were found in weight increments during the period from the beginning of gestation to the sixth month after delivery between nursing and not nursing mothers. CONCLUSIONS: The results of analysis of the dietary intake of nursing mothers indicate that the Czech recommended allowances are not met as regards energy, calcium, linoleic acid, protein and iron. Whether the intake is really inadequate or whether the recommended allowances are excessive remains an open question. PMID- 9863266 TI - [Use of EEG in electroconvulsive therapy]. AB - BACKGROUND: From the point of view of time we can divide the use of EEG in the electroconvulsive therapy into 1) screening before starting the treatment, 2) the EEG correlates of the ECT: recording of the paroxysm, immediate postparoxysmal recording, interparoxysmal recording during a series of ECT, and persistent changes after finishing the series of ECT. METHODS AND RESULTS: EEG screening has shown that the patients with abnormal EEG do not respond well to ECT. In the recording of the paroxysm EEG correlates with therapeutic effect of ECT are looked for--particularly the length of the paroxysm on the EEG, intensity and pattern of the paroxysmal activity, and the suppression of the EEG curve in the terminal part of the paroxysm. The suppression models are in the focus of interest also in the immediate postparoxysmal recording. In the interparoxysmal recordings during a series of ECT the associations of the generalised slowing in the range of the theta and delta activity with the therapeutic effect were looked for, particularly in the past. Persistent changes after finishing the ECT are usually examined especially to look if ECT does not cause persistent abnormalities of the EEG recording which would be an evidence for brain damage or for epileptic kindling. The results in this direction have been negative so far. PMID- 9863267 TI - [Must we be really concerned about amphotericin B toxicity in oncology patients?]. AB - BACKGROUND: Amphotericin B treatment in oncological patients is irrepaceable due to the high frequency of mycotic infections. From data in the literature ensues that the most serious undesirable effect of amphotericin B is nephrotoxicity, manifested by a reduced glomerular filtration and impaired tubular function (in particular the development of hypokalemia, and hypomagnesaemia). Prophylaxis of nephrotoxicity is despite major efforts unsatisfactory. In the submitted work the authors tested in a major group of patients their working hypothesis based on previous observations, that prophylactic replacement of the increasing ion losses in urine during amphotericin B treatment without waiting for a decline of serum concentrations of these ions, along with adequate hydration delays or eliminates the decline of glomerular functions. METHODS AND RESULTS: During amphotericin B therapy of 25 oncological patients renal functions, Na, K and Mg urinary excretion and the serum concentrations of these ions were followed up in detail. The urinary losses were replaced. No overall prophylaxis to prevent acute toxic reactions associated with administration of the drug was used. The mean dose of amphotericin B was 0.82 mg/kg, the mean diuresis 3662 ml/24 hours. Acute toxic reactions calling for hydrocortisone administration were observed only in 6% of the patients. During treatment the urinary K and Mg losses increased significantly and had to one replaced. There was also a significant increase of the excretory fractions of K and Na. However there were no significant changes of serum ions nor a rise of creatinine. The creatinine clearance even increased slightly though insignificantly (1.384 ml/s as compared with 1.392 ml/s). CONCLUSIONS: Consequential hydration of patients and prophylactic replacement of urinary ion losses during amphotericin B therapy are effective in preventing ion disbalances and a decline of glomerular functions. Acute toxic reactions associated with administration of amphotericin B are infrequent. PMID- 9863268 TI - Health in the Americas 1998. PMID- 9863269 TI - International classification of diseases. WHO Collaborating Centers. PMID- 9863270 TI - Blood bank situation in Latin America, 1996: serological markers for communicable diseases in blood donors. PMID- 9863271 TI - Revision of the International Health Regulations. PMID- 9863272 TI - Current controversies in carotene nutrition. AB - Most of the dietary vitamin A is derived from plant foods in the form of pro vitamin A, the carotenoids. Though in 1930 it was first demonstrated that beta carotene is the precursor for vitamin A and it is well accepted that 1 mole of beta-carotene is equivalent to one mole of vitamin A, the mechanism of conversion to vitamin A has been controversial. Some of the mechanisms suggested are central cleavage potentially yielding 2 molecules of vitamin A or excentric cleavage producing one molecule of vitamin A from beta-carotene which drastically varied the potency of carotene. A mucosal supernatant from rat intestine was shown to have beta-carotene dioxygenase activity which provided the basis for central cleavage. Many observations on enzyme activity in vitro and efficacy of carotene in vivo did not support the above findings and a re-evaluation of the whole problem was undertaken at the National Institute of Nutrition (NIN), Hyderabad. Intestinal conversion of beta-carotene to vitamin A both in vitro and in vivo in rats and in vivo in children was evaluated. A novel method of obtaining the in vivo conversion of carotene to vitamin A using the ratio of area under plasma vitamin A time curves after a dose of beta-carotene and vitamin A (> 100 micrograms) was developed in rats and later extended to children. In children a dose of 1.5 mg of beta-carotene and vitamin A was used. From these studies intestinal conversion of beta-carotene to vitamin A was found to be an enzymatic reaction involving central cleavage and which needed the presence of oxygen. The substrate was found to bind the enzyme at C-15,15'. The enzyme may be associated with inherent or contaminant enzyme which breaks of other part of the molecule released after central cleavage of carotene. The in vivo conversion of carotene to vitamin A was found to vary from 20 to 80 per cent depending on the nutritional status. Vitamin A deficiency was found to enhance both the in vitro and in vivo conversion and protein deficiency to decrease both. Thus the present results confirm the convertibility of dietary carotenoids to vitamin A and could facilitate further investigations on interactions of different dietary carotenoids on the absorption and cleavage of carotene to vitamin A in children. PMID- 9863273 TI - Bioactive phytochemicals with emphasis on dietary practices. AB - Diet can modify the pathophysiological processes of various metabolic disorders and can be an effective preventive strategy for various disease processes most of which are known to involve oxidative damage. Both nutrient and non-nutrient components of the diet have been recognized for their anti-oxidant and other potential benefits. Plant foods contain phytochemicals such as flavonoids, phenolic acids, etc., which show biological activity. Some common foods used in Indian culinary practices were assessed for their anti-oxidant, anti-mutagenic and anti-carcinogenic effects and vitamin D activity and evaluated for their plausible biological effects. Green leafy vegetables had the highest anti-oxidant activity followed by wheat and rice. Cooking decreased this activity. Eugenol, the active principle of clove, was shown to offer protection against CCl4 induced hepatotoxicity in rats. It also showed anti-peroxidative activity in addition to decrease in O2 formation. Studies on the anti-carcinogenic effect of turmeric/curcumin revealed that both are potent anti-mutagens in vivo and reduce the adducted DNA levels in liver of rates challenged with B(a)P. In another study, Syrian hamsters receiving turmeric/curcumin through diet or local paint on cheek pouch had lower tumour burden as well as adducted DNA level against 7-12 DMBA challenge. Turmeric/curcumin were found to be better anti-tumour agents when given in the post initiation phase of carcinogenesis. The beneficial effect of turmeric was found to be due to its anti-oxidant potential. Studies on humans at risk of palatal cancer due to reverse smoking showed that turmeric (1 g/day) for 9 months had a significant impact on the regression of precancerous lesions. Onion and garlic also possess anti-mutagenic principle. Further studies on the bioactive phytochemicals in plants showed that certain plants belonging to Solanaceae (Cestrum diurnum, Lycopersicon esculentum and Solanum melongena) have calcinogenic potential and vitamin D like activity. In view of the vast data on bioactive principles from plants, it is suggested that dietary prevention coupled with other life-style changes in perhaps the right answer for prevention of cancer and other chronic diseases in India. PMID- 9863274 TI - Riboflavin metabolism--relevance to human nutrition. AB - Studies in developing countries like India have revealed a very high incidence of biochemical riboflavin deficiency, particularly in women and children as judged by erythrocyte glutathione reductase activation test (EGR-AC). Riboflavin deficiency can cause conditioned deficiency of vitamin B6 and the mucocutaneous lesions observed in these two vitamins deficiencies could be due to impaired skin collagen maturity. Subclinical riboflavin deficiency impairs psychomotor function and vitamin B2 requirement may be enhanced during increased physical activity. Riboflavin status is not dependent exclusively on dietary intake of the vitamin, certain non-dietary factors can modify riboflavin status. Respiratory infection, certain diseases, drugs and hormones can influence riboflavin metabolism. PMID- 9863275 TI - Requirements of dietary fats to meet nutritional needs & prevent the risk of atherosclerosis--an Indian perspective. AB - To arrive at the fat requirements for Indians, invisible fat and fatty acid compositions were determined in commonly consumed foods. Taking into account the WHO guidelines and the invisible fat intake of Indians, the visible fat requirements according to age, physical activity and physiological status were calculated. The data from both dietary and plasma fatty acids showed that while the requirements of linoleic acid (18:2 n-6, LA) were fully met due to their high levels in cereals and many vegetable oils, alpha-linolenic acid (18:3, n-3 ALNA) intakes were low. Long-term high intakes of ALNA or long chain n-3 polyunsaturated fatty acids (LCn-3 PUFA) reduce the risk of coronary heart disease (CHD). Metabolic studies were therefore conducted in Indian subjects to investigate the effects of using fish oils (LC n-3 PUFA) and ALNA rich oils (canola oil and mustard oil, MO) in comparison to oils which have negligible ALNA (groundnut oil, GNO and palmolein, PO) on plasma lipids, essential fatty acid (EFA) status and platelet aggregation. The results showed that at a level of 6-7 energy per cent LA, about 0.2 to 0.5 energy per cent LC n-3 PUFA or 1.4 energy per cent ALNA produced anti-atherogenic effects. However, high levels of erucic acid (22:1, EA) present in MO prevented these beneficial effects. Since absolute levels as well as the ratio of n-6 to n-3 PUFA are essential for optimal health, the use of more than one oil (correct choice) is recommended. However, it is necessary to evaluate the long-term health benefits of the recommended oil combinations before permitting commercial preparation and endorsing health claims. Regular consumption of plants foods which are good sources of ALNA can also contribute to improving n-3 status. Non-vegetarians, however, have the choice of eating fish to accomplish this. PMID- 9863276 TI - Impact evaluation of iron & iodine fortified salt. AB - As a novel approach to tackle the problems of iron deficiency anaemia and iodine deficiency disorders (IDDs), which often coexist, the National Institute of Nutrition has developed iron and iodine fortified common salt (double fortified salt-DFS) as a public health measure. This salt has undergone a battery of laboratory and field tests to evaluate its feasibility for use in a national programme. The DFS is designed to provide 1 mg of iron and 15 micrograms of iodine per gram of common salt. This was made possible by the inclusion of a polyphosphate stabilizer, sodium hexametaphosphate (SHMP) at 1 per cent level. The stability of iron and iodine was found to be good up to 6 months. However, the stability of iodine depended upon the quality of the salt used for fortification. The biological effects of long-term consumption of DFS were evaluated in experimental rats and in field trials. Both iron and iodine from the salt were found to be biologically available in regenerating haemoglobin and in increasing excretion of iodine in urine. When this salt was tested in tribal villages endemic for goitre and iron deficiency anaemia, the bioresponse was good with regard to the iodine status but was not uniform in all segments with regard to iron, probably due to confounding variables. In a study carried out in residential school children where such variables did not exist, DFS was found to have significant impact on haemoglobin status in anaemic children and improved their urinary iodine excretion. The consumption of DFS for 2 yr did not have any adverse effects in school children as well as in the tribal population. Parameters related to calcium homeostasis were not altered in children receiving DFS. Histopathological examination of tissues and radiological examination of bone did not reveal any abnormality in DFS fed rats. Similarly serum and urinary parameters related to calcium and phosphorus were not altered in DFS fed rats. Therefore, DFS is presented as a feasible and effective strategy to control the double deficiency of iron and iodine in our community. PMID- 9863277 TI - Mycotoxins in foods--occurrence, health & economic significance & food control measures. AB - Mycotoxins of importance in India include aflatoxin, fumonisins, trichothecenes, ergot alkaloids and ochratoxins. The ICMR multicentric study on the occurrence of aflatoxin contamination in risk commodities namely, maize and groundnut showed that 21 per cent of groundnut samples and 26 per cent of maize samples analysed exceeded Indian tolerance limits of 30 micrograms/kg. A study on the aflatoxin intake from maize-based diets in a rural region of Andhra Pradesh showed the intakes to be in the range of 4-100 ng/kg body wt/day. Studies on the occurrence of aflatoxin M1 in milk in the southern and western regions of India indicated levels in the range of 0.05-3.0 micrograms/l. Analysis of feed samples indicated high incidence of aflatoxin B1 contamination in the groundnut cake component. Fumonisins have been shown to occur in Indian maize and sorghum. Studies showed high levels of fumonisins in rain-affected maize and sorghum consumption of which resulted in an outbreak of fumonisin mycotoxicosis in rural regions of the Deccan Plateau. A similar disease outbreak occurred in poultry due to consumption of fumonisin contaminated feed containing rain damaged maize. Biomarkers have been developed for assessing the risk of exposure for two mycotoxins viz., aflatoxin by measurement by ELISA of aflatoxin B1 N7-guanine adduct which has a detection limit of 15.6 pmol aflatoxin B1 N7 guanine; and fumonisin B1 by measurement in urine using HPLC with a detection limit of 8 ng/ml urine. Assessment of the economic implications of aflatoxin contamination showed economic losses resulting in rejection of export consignment of hand-picked-selected (HPS) groundnut and losses in the poultry industry. Approaches for prevention and control of mycotoxin contamination in foods have shown that methods involving the segregation of contaminated or mouldy grains by hand picking and density segregation resulted in a reduction of 70-90 per cent of aflatoxin and fumonisin present in the grains. While harmonization of international regulatory limits, the requirements of food producing countries needs to be recognized and realistic but not idealistic safe limits, need to be proposed. PMID- 9863278 TI - Animal models of obesity & their usefulness in molecular approach to obesity. AB - Obesity, a multifactorial nutrition disorder, is no longer the problem of the affluent West; it has been slowly gaining entry in to developing countries as well. Ever since the first demonstration of an experimental hypothalamic obese rat model, laboratory animals have been in the forefront of basic research concerned with this important metabolic disease. Apart from nongenetic models, an array of genetic murine models of obesity is now available. Over the years the obese loci in these mutants were localised, and most of them have been cloned. Among them leptin and its receptor--the first gene products to be identified, have revolutionised the field, and the possibility of a 'lipostat' mechanism operating in the body is no longer in the realm of imagination. Studies are now on, to identify the murine obese genes in the human population with a view to understand the problem and intervene therapeutically. We have recently developed a new rat model of obesity in our animal facilities, which has several advantages over the existing Western models. It is hoped, that this new model will strengthen and expand our knowledge on obesity--an interesting but complex syndrome. PMID- 9863279 TI - Diet & nutrition situation in rural India. AB - During the last 25 years, a large database has been accumulated on the diet and nutritional status of the rural population of different states of India. The National Nutrition Monitoring Bureau (NNMB) and the National Institute of Nutrition (NIN), Hyderabad have carried out extensive diet and nutrition surveys in 12 states of the country. These surveys indicate that the diets of the rural population are inadequate and deficient in most of the nutrients. There is widespread energy deficiency in the rural households. About 60 per cent of the preschool children are underweight (< median -2SD weight for age of NCHS) and 62 per cent are stunted (long duration malnutrition). About 15 per cent of the children of 1-5 yr of age suffer from short duration malnutrition (wasting). Repeat surveys showed that severe malnutrition had declined substantially in children. Similarly, there was reduction in the extent of chronic energy deficiency among the adults. However, there were no perceptible changes in the diet and nutrient intakes during the past 20 yr. PMID- 9863280 TI - A clinical, microbiological, and histopathologic study of trichostasis spinulosa. AB - Trichostasis spinulosa (TSS) is a relatively common follicular disorder that can occur on the face and trunk, especially in the interscapular area. Its cause remains unclear. We examined clinically 30 patients with TSS and follicular materials extracted from each patient were examined microscopically. Bacterial culture and skin biopsy were done in 12 and 10 patients, respectively. Periodic acid Schiff (PAS) and Brown-Brenn Gram stain were used for detection of pityrosporum (malassezia) and bacteria. The interscapular area (14/30), nose (8/30), and cheek (4/30) were common sites of TSS. Pityrosporum and bacteria in the extracted follicular material were found at the rates of 82.6% and 73.3%, respectively. In histologic examination, follicular hyperkeratosis and numerous vellus hairs enveloped within keratotic sheath were common features. Pityrosporum and bacteria were found at the rate of 70% in biopsied specimens on PAS and Brown Brenn Gram stain. In bacterial culture, Propionibacterium acne was most commonly identified in 75% (9 out of 12 patients). Pityrosporum and bacteria, especially Propionibacterium acne, were commonly found in the extracted follicular material and biopsied specimens. Thus, they may be related to the induction of follicular hyperkeratosis with retention of vellus hairs, and we suggest that these microorganisms may be one of the possible etiologic factors of TSS. PMID- 9863281 TI - Combination therapy with low-dose etretinate and eicosapentaenoic acid for psoriasis vulgaris. AB - A randomized open study was undertaken to compare the therapeutic effects between low-dose etretinate alone and low-dose etretinate combined with eicosapentaenoic acid in 40 patients with chronic, stable psoriasis vulgaris. Better and more rapid improvement was obtained with the combination therapy for 12 weeks than with low-dose etretinate monotherapy. Eicosapentaenoic acid was safe, and adverse reactions due to low-dose etretinate were mild or tolerable. The combination regimen, therefore, has a satisfactory effect on psoriasis without marked adverse reactions. PMID- 9863282 TI - Immune thrombocytopenia due to Tranilast (Rizaben): detection of drug-dependent platelet-associated IgG. AB - Tranilast (Rizaben)-induced thrombocytopenia occurring in a 17-year-old man was reported. After withdrawal of the drug, he recovered within a week with oral prednisolone administration. Serological examination revealed no anti-platelet antibody, but platelet-associated IgG (PAIgG) was found. After incubation of peripheral blood of the patient with the drug in vitro, the level of PAIgG was significantly increased. These findings suggest the presence of a drug-dependent anti-platelet IgG in the patient's serum. This is the first report of immune thrombocytopenia caused by Tranilast. Our method for detecting drug-dependent platelet antibody in vitro is safe and useful for diagnosing drug-induced thrombocytopenia. PMID- 9863283 TI - Effect of nonenzymatic glycosylation on the titers of circulating autoantibodies in pemphigus and pemphigoid. AB - Hyperglycemia is observed in some patients with autoimmune bullous diseases complicated by diabetes mellitus or treated with systemic corticosteroids. High concentrations of glucose can react with various proteins and change their structural and functional properties. We previously reported that nonenzymatic glycosylation of antibody can impair antigen-antibody binding. We ascertained whether glycosylation of autoantibody decreases the autoantibody titer by examining 30 sera from patients with pemphigus and pemphigoid. Nonenzymatic glycosylation in the physiological range was induced by incubation of sera with 1650 mM D-glucose at 4 degrees C for 7 days. The titers of sera were determined by indirect immunofluorescence (IIF). In all cases, the immunofluorescence intensity of glycosylated sera was weaker than that of nonglycosylated sera. Glycosylated sera showed a lower antibody titer by 1 doubling dilution in 18 out of 30 cases, compared with nonglycosylated sera. The ten BP patients' sera were also analyzed by immunoblotting for reactivity with the BP180-GST fusion proteins, S delta 1 and 4575. All BP sera reacted with S delta 1, and 5 out of 10 BP sera reacted with both S delta 1 and 4575. In all the sera that reacted only with S delta 1, the glycosylated sera showed a 1 doubling dilution decrease in autoantibody titer. Interestingly, in 4 out of 5 sera that reacted with both S delta 1 and 4575, there were no differences in the antibody titer between glycosylated and nonglycosylated sera. These results indicate the possibility of a false decrease in autoantibody titers of sera from patients with autoimmune bullous diseases complicated with hyperglycemia. Although the false decrease in titers of autoantibodies induced by nonenzymatic glycosylation is not dramatic, it must be considered in order not to underestimate the disease activity of pemphigus in such cases. PMID- 9863284 TI - A case of calcinosis universalis successfully treated with low-dose warfarin. AB - A 27-year-old male patient with calcinosis universalis resulting from dermatomyositis was successfully treated with low-dose warfarin. On his trunk and extremities, there were many subcutaneous calcified nodules, and knee flexion was difficult. After oral warfarin therapy for three years, the calcified nodules became smaller, and the knee mobility improved. His serum vitamin K level was abnormally high, decreased just after starting warfarin therapy, and then remained within the normal range. Since vitamin K has been known to play an important role in the Ca2+ binding process in bones or tissues, we suggest that this therapy is effective in reducing subcutaneous calcification through the vitamin K cycle. PMID- 9863285 TI - Phacomatosis pigmentovascularis type IIb associated with Sturge-Weber syndrome and pyogenic granuloma. AB - A case of phacomatosis pigmentovascularis (PPV) in a 6-year-old girl with Sturge Weber syndrome, pyogenic granuloma, and other complications is described. It is relatively rare that a complete form of Sturge-Weber syndrome was associated with PPV. A review of the literature on PPV, focusing on total number of reported cases and etiological speculations, is presented. To our knowledge, a total of 118 cases of PPV, including the present one, have been reported to date. Regardless of many speculations, the true etiology remains unknown. The average "density" of mast cells (MCs) per mm2 appearing in the central region of the pyogenic granuloma was calculated to be 86.3/mm2 and that in the adjacent nevus flammeus was 37.9/mm2. The "density" of mast cells in pyogenic granuloma separately calculated from ten other cases was 105.5 +/- 28.6/mm2 (mean +/- SD), compared with that in normal skin, 6.85 +/- 4.9/mm2 (n = 20). There was a significant difference between the two, indicating that MCs are closely associated with angiogenesis in pyogenic granuloma. PMID- 9863286 TI - Poikiloderma-like cutaneous amyloidosis in an ethnic Chinese girl. AB - Primary cutaneous amyloidosis is the deposition of amyloid in the skin without involvement of internal organs. It is easily diagnosed when presented in its typical manifestation. Atypical or rare clinical presentations can pose diagnostic difficulties. Poikiloderma-like cutaneous amyloidosis (PCA), a rare variant of primary cutaneous amyloidosis, was first reported in the literature in 1936 (1). It is characterised by: 1) poikilodermatous skin lesions; 2) lichenoid papules; 3) cutaneous amyloid deposit in the pigmented and lichenoid lesions; 4) light sensitivity; 5) short stature; and 6) other features such as blister formation or palmoplantar keratosis. Ogino coined the term PCA syndrome when these unusual features present early in life (2). We report a 26-year-old Chinese woman who presented with poikilodermatous skin lesions and was misdiagnosed as poikiloderma atrophica vasculare (PAV) on the basis of clinical appearance without any histological proof. The diagnosis of PCA was made after skin biopsy which showed amyloid deposits in the skin. This condition can easily be confused with other true poikiloderma skin diseases. Histology is important in confirming the diagnosis. PMID- 9863287 TI - A case of congenital infantile fibrosarcoma of the right hand. AB - Congenital infantile fibrosarcoma is a rare soft-tissue malignant tumor that usually presents as a mass involving the arm or leg. We report a case of congenital fibrosarcoma on the right hand of a newborn infant. The patient was a 12-day-old male. From the time of delivery, he had a red tumor on his dorsal right hand. The tumor was initially associated with bleeding within a hemangioma and compressed. The course was observed. It appeared that the tumor might be malignant because it increased in size to 10 x 8 x 3 cm, and ulceration and bleeding appeared on the surface after 10 days. A biopsy suggested a fibrosarcoma. The tumor was resected 32 days after birth at the narrow margin in consideration of preservation of function. However, a recurrence was observed under the graft after 2 weeks and another resection was performed. Because the tumor had passed through the interosseous muscle and invaded the palm of the hand, the middle finger, ring finger, and little finger had to be amputated. One year and 7 months postoperatively, no recurrence or metastasis has been observed. PMID- 9863288 TI - Eosinophilic pustular folliculitis (Ofuji's disease) in myelodysplastic syndrome. AB - We describe a case of eosinophilic pustular folliculitis (EPF, Ofuji's disease) in a 12-year-old male who suffered from myelodysplastic syndrome. Bone marrow study revealed an increase in the eosinophil cell line without peripheral blood eosinophilia in our case. We suggest that the immunologic abberations ascribed to myelodysplastic syndrome and the increase in the eosinophil cell line in the bone marrow might play roles in the development of EPF in our case. PMID- 9863289 TI - Erythema dyschromicum perstans: report of a new case and critical review of the literature. AB - Erythema dyschromicum perstans (EDP), described by Convit et al. in 1961, is a rare dermatosis. Its relationship with ashy dermatosis (AD), described by Ramirez in 1957, is still a matter of debate. We report a typical case of EDP. The patient, of North African origin, had a dyschromic (hypo- and hyperpigmented) eruption on the chest and limbs for 2 years. The lesions were occasionally surrounded by a papular border which spread slowly and centrifugally. Histological examination showed a lichenoid infiltrate. A carcinoma of the lung was simultaneously discovered. No treatment was given, EDP is infrequent and often considered identical to ashy dermatosis in the literature. However, the clinical aspects of the two diseases differ. The main features of these two diseases are reviewed and compared on the basis of a literature review. We conclude that EDP and AD are distinct clinical entities. PMID- 9863290 TI - Fibromyxoma of the skin. AB - A patient presented with a solitary large subcutaneous tumor homogeneously composed of loose fibroblasts interspersed with abundant mucinous material. The literature was reviewed, and the origin, pathogenesis and clinical course of this rare neoplasm were briefly discussed. PMID- 9863291 TI - Figurate purpuric eruptions on the trunk: acetaminophen-induced rashes. AB - A 23-year-old man had recurrent erythematous purpuric patches in a transverse linear arrangement on his back and arms. One week prior to a recent episode, he took acetaminophen for 3 days. Four months earlier, he developed the same purpuric lesions at/around the same anatomical site when he took acetaminophen for pain-relief. A biopsy specimen showed capillaritis and extravasation of erythrocytes in the papillary dermis. A provocation test with acetaminophen confirmed these lesions as drug-induced rashes. The uncommon topographic and morphologic features of the purpuric lesion in this patient might be considered as an unusual expression of pigmented purpuric dermatosis caused by acetaminophen. PMID- 9863292 TI - Numerous hypopigmented patches associated with atopic dermatitis. PMID- 9863293 TI - [Pain in hernia has not only mechanical basis. Pharmacological treatment can open new possibilities]. PMID- 9863294 TI - [Suicide--a neglected public health problem]. PMID- 9863295 TI - [Can ethical reconsideration result in increased number of organ donations?]. PMID- 9863296 TI - [Uncertain timing trend in connection with cadmium exposure]. PMID- 9863297 TI - [Limited effect of antidiabetics on type 2 diabetes]. PMID- 9863298 TI - [A private hospital listened to the relatives and survivors from the ship Estonia. The Ersta hospital voluntarily took over from the crisis intervention groups]. PMID- 9863299 TI - [New information concerning pain caused by herniated disk and sciatica. Exposure to disk tissue sensitizes the nerve roots]. AB - Sciatica has long been considered to be solely due to mechanical deformation (compression) of a spinal nerve root by herniating disc tissue. However, recent experimental findings have demonstrated that, even in the absence of mechanical insult, nucleus pulposus-related substances may not only induce significant structural and functional injury to the adjacent nerve root, but also sensitise the nerve root, producing pain in the event of subsequent root compression. Although neither the specific mechanisms nor the active nucleus pulposus-related substances causing pain have yet been identified, the recent findings suggest that the future may well bring new pharmacological treatment alternatives for sciatica and disc herniation. PMID- 9863300 TI - [Treatment of multiple sclerosis--1. New drugs may be effective but there still are frequent relapses]. AB - Multiple sclerosis (MS) is a demyelinating, central nervous system disease, of putative autoimmune pathogenesis. Although no effective pharmacological therapy has been available for this often disabling disease until recently, several studies have now confirmed that subcutaneous or intramuscular administration of beta-interferon may reduce the frequency and severity of relapses in relapsing MS, and may also inhibit disease progression. Studies are under way to determine the possible efficacy of beta-interferon during the progressive phase of the disease. Three beta-interferon formulations are currently available in Sweden. Another drug, glatiramer acetate, also shown to have some effect on the disease course, is expected to be registered for use in Sweden shortly. PMID- 9863302 TI - [The American wheelchair was not approved by the king]. PMID- 9863301 TI - [Treatment of multiple sclerosis--2. Application of a new immunological basic research is close]. AB - Based on continuous advances in our knowledge of the pathophysiological mechanisms involved in the development of multiple sclerosis and other putatively autoimmune diseases, several new immunological treatment strategies are currently being tested in different parts of the world. In principle, these strategies are designed to interfere with three steps in the presumed pathogenic immunological process: inhibition of lymphocyte recruitment to inflammatory foci in the central nervous system; blocking of antigen-specific activation; and interference with immunological effector mechanisms. Preliminary results are available for some of these treatment strategies. PMID- 9863303 TI - [Sperm donors in Umea after 1985: increased number of sperm donors and higher pregnancy rate]. PMID- 9863304 TI - [Eye complications in diabetes. According to new criteria patients with diabetes should have ophthalmological examination at the time of diagnosis]. AB - Diabetic retinopathy is still the most common cause of blindness among people under 65 years of age in the Western world. Early detection of sight-threatening retinopathy is vital, as in many cases it can be treated successfully with photocoagulation. Accordingly, screening programmes using fundus photography have been introduced in many countries, and in Sweden repeat examination at two-year intervals has been recommended. Since up to 30 per cent of patients with type 2 diabetes (according to the previous criteria) have been shown to manifest diabetic retinopathy at diabetes diagnosis, an initial ophthalmological examination at diagnosis is recommended. The new diagnostic criteria for diabetes, according to which patients with fasting blood glucose levels of 6.1 (6.6 mmol/L are also classed as diabetic, will increase the number of diabetic patients. Until such information becomes available as to the prevalence of sight threatening retinopathy at diabetes diagnosis in that subgroup, it is suggested that type 2 diabetes patients should continue to undergo retinal examination at diabetes diagnosis. PMID- 9863305 TI - [Incidents at a department of anesthesiology: communication problems and deviations from the routines are the most common errors]. PMID- 9863306 TI - [Attitude of personnel toward incidents at a department of anesthesiology: reporting is important for both patient safety and occupational environment]. PMID- 9863307 TI - [Postpartum atony. A simple method prevents life-threatening hemorrhages]. PMID- 9863308 TI - [Further studies are required: in spite of positive effects of statins many patients still develop myocardial infarction]. PMID- 9863309 TI - [Increasing frequency of suicide among adolescents? Statistics should be interpreted with caution]. PMID- 9863310 TI - [Suicide epidemic after publication of "The Young Werther"? The most-read book in Europe was considered life-threatening]. PMID- 9863311 TI - [Mass media are responsible for suicide prevention]. PMID- 9863312 TI - [The health care system in the USA does not welcome patients with "minor complaints". It takes days before they are seen by a family physician]. PMID- 9863313 TI - [Reaction of thymic mononuclear phagocytes in rats 2 months after pinealectomy, ultrastructural-morphometric analysis of interdigitating cells]. AB - The effects of pinealectomy on the dynamics of morphometric changes in interdigitating cells(IDC) of rat thymus were investigated. The experimental animals were divided into two groups: an experimental one and the control group. Animals from the first group were subjected to pinealectomy while the second group were treated in the same manner, but without the removal of the pineal gland. All the animals were sacrificed 60. days following the surgery. The thymus tissue were fixed by means of immersion and prepared for transmission electron microscopy. Using Weibl's multipurpose test system and multilavel sampling technique on electron micrographs, the nuclear volume density(Vvn) and cytoplasmic volume density(Vvc) were calculated. At the different magnification level established the surface density(Sv) of nucleoid and cell membranes as wel as surface density of rough endoplasmic reticulum and mitochondrial membranes (Svm) and their volume density. Our analysis has confirmed statistically significant increase in Vv of the nucleus of pinealectomized rats. The average volumen of IDC has been also increased after pinealectomy as well as Vv, Nv and Sv of the mitochondria. The present findings seem to support the concept of inhibitory action of pineal gland on thymus IDC. PMID- 9863314 TI - [Characteristics of arterial blood pressure responses in the treadmill stress test in patients with post-traumatic stress disorder during the war in Sarajevo]. AB - The health problems associated with PTSD include multiple physical symptoms and signs. It was observed that PTSD was often accompanied with cardiovascular problems. The aim of this study was to estimate the characteristics of arterial blood pressure response on stress exercise in PTSD patients who were forced to stay in a war zone of Sarajevo. This study was carried out in 1995, during the period of intensive war activities in Sarajevo. Exercise stress testing was performed on treadmill, under the Bruce protocol in age matched 9 male PTSD patients and 14 healthy young volunteers (age 18-23 years). Arterial blood pressure was obtained by the cuff method in rest, at the last minute of each exercise stage and four times in the recovery period (1, 3, 6 and 9 minutes). Our results did not confirm previous reports related to the resting values of arterial blood pressure in PTSD patients. In rest, there were no significant differences in systolic, diastolic and mean arterial pressure between PTSD and control group. Systolic blood pressure (SBP) increased during exercise in both groups, especially at advanced exercise stages. Maximal mean SBP values were 222.00 +/- 24.80 mm Hg (control group), and 179.80 +/- mm Hg (PTSD). Exaggerated systolic blood pressure response (over 200 mm Hg) was observed only in the control group. This is probably due to physiological hyperreactive with enhanced sympathetic activity in response to stress caused by war. The luck of hyperreactive response in PTSD patients could be explained by diminished sympathetic influence on systolic blood pressure probably caused by psychoactive medication. PMID- 9863315 TI - [Serum cyclosporine levels in patients after kidney transplantation]. AB - In this study we have analyzed the cyclosporine levels in patients after kidney transplantation. The cyclosporine levels are determined by using fluorescence polarization immune-assay and monoclonal whole blood test. We analyzed nine patients with transplanted kidneys who were on specific immunosuppressive cyclosporine therapy. The cyclosporine determination was performed in these patients three times in the span of 14 to 29 days. The cyclosporine levels in seven patients were in reference interval between 100 and 300 ng/ml of whole blood. In the first determination the values in two patients were out of reference interval, one value was lower, and the other one was higher then reference interval. In second determination after dose correction in patients who had lower cyclosporin level the new level was normal and the other patient cyclosporine value was lower then in the first determination, but still a little higher then reference values. PMID- 9863316 TI - [Amplification of specific cytomegalovirus DNA sequences using the polymerase chain reaction]. AB - Highly sensitive and specific laboratory methods are essential for an early diagnosis of Cytomegalovirus (CMV) infections. Neither method of tissue culture nor CMV serology methods can serve for that purpose but they are useful for the postsymptomatic diagnostics. During the infection, CMV stays in body fluids and the appearance of clinical signs is strictly correlated to viremia. Early detection is of a great therapeutical importance. It is achieved by sensitive and specific Polymerase chain reaction (PCR) method, which proved extremely efficient and not time consuming. We used specific nested CMV-DNA-PCR (gB target) method for the analysis of 46 plasma specimens in order to determine a presence of highly conservative nucleotide sequences of gB CMV-gene. There were 17 positive samples. Our results showed that this method is useful for early detection of CMV DNA in presymptomatic diagnosis of seronegative CMV infected patients. PMID- 9863317 TI - [Arthroscopic treatment of fractures of the tibial plateau]. AB - The author presents a new operative approach (arthroscopic assisted treatment and fluoroscopy). It is a minimal invasive surgical procedure which consists of the reposition and fixation of tibial plateau fractures type B/ASIF classification. In the period 1995-1997 on Traumatology Clinic, Clinical centre in Sarajevo 10 patients with tibial plateau fractures have undergone a surgical treatment (9 patients with lateral, 1 patient with medial). Seven patients have an isolated injury and 3 patients have a complex injury. After surgical procedure, the final result was evaluated according to Final Score Scale (FSS) by Lysholm and Radiological Score Scale (RSS) by Jarvinen-Kannus and it was excellent. In the postoperative period an early movement procedure was applied and deep venous thrombosis (DVT) was prevented by A-V impulse system and required heparin administration. PMID- 9863318 TI - [Alcohol as a cause of suffering in traffic accidents]. AB - Alcohol is a cause of traffic accidents quite often. It is a reason for 18% of traffic accidents. Beside the drivers, the assistant drivers and pedestrians also get killed quite often. Children get killed because of the alcoholism of adults (18.7%). In the first place, male drivers consume alcohol beverages. Females, as assistant drivers and pedestrians get killed less often because of alcoholism then males. PMID- 9863319 TI - Trichomoniasis of the breast diseased by fibrocystic mastopathy: pathogenic rather than saprophytic relationship (Trichomonas in fibrocystic mastopathy process). AB - Because of the striking similarity between histopathological pictures of chronic trichomonal cervicitis uteri, the tissue reaction after subcutaneous inoculation of Trichomonas culture in experimental animals and female breast diseased by fibrocystic mastopathy, the detection of Trichomonads was undertaken in surgically removed diseased breast parts. In 12 FCM patients, subjected to segmental breast resection, the imprint smears were prepared from dissected specimens and after supravital staining immediately examined by light microscopy. The mucous content from dilated ducts was inoculated in the culture media. The dissected tissues have been further histologically analysed by standard method. In nine out of 12 examined FCM cases the direct microscopy revealed aflagellary, pseudocystic, leucocytoid form of Trichomonads. The cultures were positive in 4 cases: in 3 patients Trichomonas tenax was identified and in the last one T vaginalis. Histopathological findings in all 12 examined cases have shown the changes characteristic for FCM. On the basis of the accumulated knowledge about pathogenic capacity of Trichomonads, it can be with great certainty claimed that these protozoa, even in their pseudocystic form are able to cause the all appearances characteristic for FCM. This first report about Trichomonas infection in the middle of FCM process gives the unexpected hope in solving of its etiology and new insight into antitrichomonal host reaction which is frequently associated with epithelial dysplasia and unrarely with precancerous lesions as earlier observed in cervix uteri. PMID- 9863320 TI - [Quality assessment in primary health care]. AB - In this research paper we gave a model with a scale for assessment of health services quality in the primary health care. The model of assessment of quality is based on elements of quality--structure, process and outcome of health care. For every quality element defined indicators and variables with numeric values are shown. The proposed model was tested on the sample of hundred check ups of children aged from 0 to 1 year. The source of data was the health record, and data were collected through questionnaire for assessment of medicine team procedures. The outcome is measured through questionnaire about parents' satisfaction with services delivered. The results of research showed that the health record is only partly suitable to obtain data about contents of doctors work. Total quality of health care is estimated as lower values. The reason lies in the lower level of the content and quantity of check ups, and insufficient education of health professionals. The suggested model and evaluation scale are suitable for implementation in our health care system. The assessment of quality of work is compulsory in further evaluation of the system, regarding the obtained results. PMID- 9863321 TI - Could the community social alarm system serve also as a primary health care service? AB - The history of calls to the first community social alarm centre in Ljubljana and the service subscribers records were analysed with the aim of finding out how the service helps in emergencies when calls related to health problems are received. The network, being a pilot project in Slovenia, was established in 1992 primarily to offer social services to elderly and disabled people living in their own home environments. The article highlights the role of the network in solving the medical problems of the users of the system. Among the 18,500 calls received in four years at the centre, 395 (2.1%) of them were health-related. An additional 24 emergency calls (0.2%) of them were received from non-subscribers to the centre using an "ordinary" telephone. Although the absolute number of health related calls is low, the network plays an important role in preventive primary health care aimed at the observed population groups. The network meets most of the specific technical and organisational requirements for a community health care emergency call system. The described system suitability and low percentage of calls related to health problems suggest that there is no need for two parallel call systems: one to cover social and the other one to fulfil emergency calls for medical help to elderly and disabled at their homes. From a telematic point of view it is important that a telematic device--an emergency call unit- has entered an elderly or disabled person's home and been accepted. The unit could be a basic communication device for different telemedical applications. PMID- 9863322 TI - [Ewing's soft-tissue sarcoma--case report]. AB - This paper reports a very rare case of 5-years old girl with a tumor of the right kidney and metastatic changes in the inferior vena cava and right heart. Following the nefrectomy Wilms' tumour diagnosis was established. There was no an adequate answer on chemotherapy according to SIOP i.v. protocol. Three months later metastatic changes spreaded to lungs, pelvic bones and distant lymphonodes of the neck and axillas. The patient in a very severe condition was evacuated to England (Alder Hay Children's Hospital, Liverpool). After numerous tests the diagnosis of Wilms's tumor was excluded and diagnosis of Ewing's sarcoma (soft tissue) was established. PMID- 9863323 TI - [Neurosarcoidosis presenting with transitory neurodeficit and generalized epileptic seizures associated with migraine]. AB - Sarcoidosis is a multi-system disease of unknown etiology with abnormal immune response. Neurosarcoidosis affects 3 to 5% of all patients with sarcoidosis. A case of the patient with neurosarcoidosis, has been presented in this article. He has had leptomeningeal involvement and white matter lesions of the left cerebral hemisphere and clinically, transient neurological deficit, generalised epileptic seizures and migraine headache. PMID- 9863324 TI - [Organization of radiologic services in Sarajevo during the war (1992-1995)]. AB - The organization and functioning of radiological diagnostic services in Sarajevo during the three and a half years of aggression are presented. The sudden and brutal aggression, all kinds of blockage, and total unreadiness of the services caused many problems regarding the number, age structure and qualification of the staff, expended and technically defective equipment lacking spare parts and shortage of electric power, water, heating, of X-ray films, chemicals, contrast agents, sanitary, and all other kinds of material. In spite of all these problems and the necessary narrowing of indications, from April 1992 till September 1995, 340,079 radiological examinations were performed at the Institute of Radiology and 82,924 in the State Hospital, respectively. The majority of examinations was done on the muscular-skeletal system and the thorax. All radiological techniques like classic roentgenology, ultrasound examinations, computerised tomography and digital methods were used. The organisation employed in both institutions proved justified, rational and effective. It can be concluded that in this difficult period the radiological diagnostic services in Sarajevo fulfilled what was demanded of them. Regarding the extremely difficult circumstances, their contribution could also be deemed unexpected. PMID- 9863325 TI - Genetics of mood disorders--an overview. Part two. AB - The present article reviews the recent molecular findings in mood disorders. Results of linkage and association studies are discussed in regard to the main limitations of these approaches in psychiatric disorders. On the whole linkage and association studies contributed to the localisation od some potential vulnerability genes for Bipolar disorder (BP) on chromosomes 11, 4, 21 and X. The hypothesis of anticipation in mood disorders is also considered in light of interesting results with trinucleotide repeat expansions. PMID- 9863326 TI - Iodized salt--a constant concern of society. PMID- 9863327 TI - [Acute phase proteins in acute pancreatitis]. AB - INTRODUCTION: Most attacks of acute pancreatitis are self-limiting, but in 10-20% of cases, however, severe diseases with systemic complications develop. During the last few years, it has been recognized that acute phase proteins have an important role in the pathophysiology of acute pancreatitis. The present study examines the value of C-reactive protein, alpha-1-antitrypsin and orosomucoid in the assessment of severity of acute pancreatitis. MATERIAL AND METHODS: 150 adult patients suffering from acute pancreatitis by Mayer's clinical criteria (10) were divided in two groups. The first one (n = 50) consisted of patients with severe form of the disease and the second (n = 100) of patients with a mild form of acute pancreatitis. Acute phase proteins (C-reactive protein, alpha-lantitrypsin and orosomucoid) were determined quantitatively in both groups on the 1, 2, 3, 7 and 14th day of the disease onset. RESULTS: Increase in C-reactive protein values was observed in both groups. There was statistically significant greater increase in C-reactive protein in patients with severe acute pancreatitis than in those with mild form of the disease. C-reactive protein values fell slowly in all patients. Serum alpha-1 antitrypsin values were less increased than C-reactive protein values. There was a greater increase in patients with severe form of disease. The increase of orosomucoid was seen only after the third day of the disease onset and there was not a significant difference in values between the two groups. DISCUSSION: One of the most important problems in treating patients with acute pancreatitis is to detect patients with a severe form of the disease as early as possible, so that adequate treatment can be started immediately. The severity of acute pancreatitis is graded by Ranson and Imrie scores, but they request 48 hours for prognosis to be defined. Despite intensive research, no single laboratory test or pathophysiologic parameters have been found to accomplish early diagnosis. Recent studies suggest that single biologic markers such as acute phase proteins may soon allow a simple and early assessment of the prognosis. CONCLUSION: This study suggests that a C-reactive protein is a good early marker for the severity of acute pancreatitis. The high increased levels at the beginning point to serious course of disease in future. The increase of alpha 1-antitrypsin is of a similar importance, while the increased orosomucoid, appearing only after the third day, is of no importance in relation to the prognosis of the disease. PMID- 9863328 TI - [Submandibular secretion of immunoglobulins IgA and IgG in peripheral paralysis of the facial nerve]. AB - INTRODUCTION: In all lesions of the facial nerve suprachoroidally localized, and due to disturbance of parasympathetic and sympathetic component, there comes to qualitative and quantitative disorders of the secretion of submandibular salivary gland. Glandular immunoglobulins IgA and IgG are the secretion of the specific plasma cells in the interstice of this gland. The mechanism of the secretion of immunoglobulins is not sufficiently clear, but it is certainly under the direct neurogenic control, since the disorders of the secretion emerge after the denervation of the submandibular salivary gland. The aim of the study was to prove the direct relation between the degree of submandibular immunoglobulin secretion IgA and IgG, and the degree of the lesion of the facial nerve U which is vitally important in the clinical estimation of the peripheral paralysis of this nerve. MATERIAL AND METHODS: In 35 patients with peripheral idiopathic facial nerve paralysis, the quantity of the secreted immunoglobulins IgA and IgG was examined by laser nephelometar BLN, Module 3. The quantity of the secreted immunoglobulins IgA and IgG (mg lit) in the saliva of the paralysed side was indirectly compared to the secreted immunoglobulins of the healthy, i.e. control side. The examination was performed three times: a) after the appearance of the disease, in the first 30 days; b) two to three months later; c) after six to twelve months. RESULTS: The quantity of the secreted immunoglobulins is significantly higher in the saliva samples taken from the paralysed side (9.50U204.77 mg lit), in comparison with the samples taken from the healthy side (9.50U70.36 mg lit). In the group of patients with favourable results and significantly higher secretion (p 0.01) normalization occurred in the final period of observation. In patients with unfavourable results the difference in secretion was continuously present (p 0.05) (table 1). DISCUSSION: In the lesions of the facial nerve suprachoroidally localized, there comes to disorder concerning the secretion of immunoglobulins IgA and IgG by submandibular salivary gland, which can be applied in the estimation of the degree of paralysis and the prognosis of the final result. CONCLUSION: The results of the research show that in the peripheral idiopathic facial nerve paralysis, there comes to increased secretion of immunoglobulins IgA and IgG in submandibular gland, at the paralysed side. In the patients who, during the paralysis, show quicker fall and normalization of the previously increased quantities of immunoglobulins, the recovery of the motor function of the facial nerve comes more successfully and more certainly. The degree of the secretion of immunoglobulins IgA and IgG can be used for the estimation of the severely of the pathological process in the suprachoroidal part of the nerve, and it can be used as a reliable parameter for the prognosis of the paralysis outcome. PMID- 9863329 TI - [Short bowel syndrome--surgical aspects]. AB - INTRODUCTION: Long-term survival after massive intestinal resection is now possible with parenteral nutritional support. The expense, morbidity, and inconvenience of this therapy, however, have led to continued interest in alternatives for the treatment of the short bowel syndrome. Patients with short bowel require a multi disciplinary approach over a prolonged period. HISTORICAL CONSIDERATIONS: The history of small bowel transplantation started in 1959 when Lillehei showed that autotransplantation of the small intestine in a dog was feasible. From 1964 to 1971, 7 attempts of small bowel allotransplantations in humans have been reported. All 7 patients died. DEFINITION: Short gut syndrome is a malabsorptive condition occurring after significant loss of intestinal absorptive capacity. The clinical syndrome is manifested by malnutrition, steatorrhea, weight loss, and diarrhea due to decreased absorptive capacity. ETIOLOGY: Etiologic factors leading to the short gut state include necrotizing enterocolitis, midgut volvulus, trauma, embolic phenomenon, and Crohn's disease. PATHOPHYSIOLOGY: Intestinal failure is the end result of several complex interacting mechanisms related to: reduced enterocyte mass, short small bowel length with consequent reduced mucosal contact time for absorption, massive proximal loop dilatation with poor propulsion, and intraluminal stasis and bacterial overgrowth lead to bacterial translocation to the liver systemically. MANAGEMENT: Patients with short bowel must be totally or partly supported with intravenous nutrition until enteral absorption can sustain survival and growth. Autologous bowel reconstruction attempts to reconfigure the residual bowel to eliminate negative factors of bowel dilatation and stasis, and redistribute the absorptive mucosa to enhance the adaptation response. Several procedures have been suggested to: prolong transmitting time and increase mucosal contact time, enhance absorption by bowel tailoring and bowel lengthening, and increasing the Enterocyte mass. CONCLUSION: Autologous gastro-intestinal reconstruction is still in its infacny with prospect of new and different concepts for the future. PMID- 9863330 TI - [Leptin and human reproduction]. AB - Leptin is a secretory product of adipocytes. It has been suggested that leptin acts as an afferent satiety signal to the brain modulating the expression of the orexigenic hypothalamic peptide, neuropeptide Y (NPY). Therefore leptin can be regarded as a marker of the nutritional status of the body. It was proposed that human obesity may result from a central resistance to leptin due to different pathophysiological mechanisms: saturation of the leptin transport into the cerebrospinal fluid of the obese subjects, abnormalities in the hypothalamic receptor for leptin, or post-receptor transduction mechanisms. It was shown that circulating leptin levels in humans significantly correlate with the body mass index (BMI). Although most studies point to white adipose tissue as a primary source of leptin there is still some uncertainty towards the relative expression of leptin between various body fat compartments. LEPTIN AND ONSET OF PUBERTY: Studies on animal models recognized various metabolic candidates for modulation of GnRH neuronal activity. It was supposed that mild changes in the body's metabolic status can serve to regulate the central drive to the reproductive axis. It is likely that leptin can serve as a "metabolic cue" that transmits signals of those mild metabolic changes towards activation of the GnRH neuronal system at the end of the prepubertal period. On the other side there is a possibility of altered leptin pulsatility during prepubertal period that can consequently influence hypothalamus and GnRH neuronal system. LEPTIN AND SEXUAL DIMORPHISM: Leptin levels in humans are similar in both sexes during the prepubertal period. During puberty leptin has a tendency to decline in boys and to remain constant in girls. Puberty is also characterized with a similar circadian rhythm pattern between sexes whil girls express different pulse characteristics. It seems that sexual dimorphism is established in early phases of human development. There is a possibility of sex steroid influence on such sexual dimorphism. LEPTIN AND REPRODUCTIVE FUNCTION: It was shown that administration of recombinant leptin to ob ob mice could restore fertility in these infertile animals. There is certain difference in leptin levels according to the phase of the menstrual cycle. It was shown that leptin peak is in the luteal phase of the cycle and that correlates to the maximal progesterone level. It is possible that leptin could directly influence ovary and that disruption of such an effect could play a role in menstrual irregularities in both obese and mal nourished women. This could even become a pathophysiological mechanism in women with polycystic ovary syndrome (PCOS). It was supposed that leptin resistance could be involved in infertility impairment of the obese women with PCOS. Leptin increases during pregnancy. Appearance of placenta as a new, nonadipose source of leptin production, increases a possibility of different leptin mRNA expression through gestation. PMID- 9863331 TI - [Classification of laser irradiation and safety measures]. AB - The use of lasers in medicine and especially surgery is rapidly expanding in many disciplines from clinical laboratory to the office practice and operating room. It is essential that users of this powerful tool have knowledge of their potential hazards and the measures to protect patients and personnel against injuries or undesired effects. Below, we have included information about the way lasers are classified; the development of protective standards; the current status of protection standards that apply to lasers, especially those used in medicine/surgery; the specific kinds of hazards associated with medical/surgical applications; and the measures by which hazards have been controlled. Since laser technology is still a young field, it is likely that problems unknown at present will occur and methodologies for controlling hazards will evolve. The American National Standards Committee produced the first consensus standard Z136.1 in 1973. The Standard was revised in 1976 to accommodate differences in biological effects for different wavelengths in the visible spectrum. The ANSI Standard has been revised again in 1980, and currently (1984) there are two additional standards in preparation, Z136.2 and 136.3, which treat the safe use of light emitting diodes and the safe use of lasers in the health care environment, respectively. Most surgical and medical lasers are Class III or IV. Some lasers have a Class IV therapy level beam plus a Class I or II alignment beam. When using lasers, it is possible to generate incandescence or fluorescence in an irradiated object. This can occur even with protective eyewear, because the correlated radiations are usually of a different wavelength. Generally, this should not be a problem when beams are directed at biological material. However, hazard could be caused by lasers designed to produce fluorescence. Control of correlative radiation in a laser system is required in the federal regulations. Hazards of lasers may be grouped as those to the eye, skin and associated hazards, fire, x-rays, electrical, fumes, toxic materials, etc. Effects on tissue are governed by the following factors:--the energy or power density of the beam;- the absorption in tissue at the laser wavelength;--the time the beam is held at a given area.;--the protective effects of heat removal by thermal conduction and by circulation. Eye hazards include thermal burns or acoustical disruption (shock waves) from high-powered or high-energy beams in the visible and near infrared wavelengths. Direct beam exposure or specular or diffuse reflaction from these very high-power lasers can also cause injuries to other parts of the retina. For example, beams can directly penetrate through the sclera and cause retinal injury. Near ultraviolet (less than 400 hm) and far infrared (.3000 hm including CO2 lasers) can cause moderate to severe corneal burns. Far ultraviolet (200-315 nm), mid infrared (1400-3000 nanometers) can cause welders' flash or snow blindness and chronic exposure could cause cataract, and exposure to ultraviolet rays may be carcinogenic. For CO2 lasers, the far infrared radiation is attenuated by plastic goggles, or by glasses, or quartz. Other eyewear with special filters is used for different lasers. The whole personnel who may be exposed to direct beams, specular reflections, and many times diffuse reflections must wear protective eyewear. In all cases the surgeon and others viewing the procedure through the endoscope need glasses or suitable protective lenses installed in the endoscope. Persons who are not viewing the beam may not need to wear protective glasses with the same level of optical density. Glasses may be selected to provide protection for lasers operating in the visible wavelength to the point where the normal aversion response could protect the individual. PMID- 9863332 TI - Induction of labor by endocervical application of prostaglandins and intravenous infusion of oxytocin in postterm pregnancy. AB - The study compares two contemporary modes of labor induction: intravenous application of Syntocinon and endocervical application of prostaglandins and examines their efficacy and safety. Ninety women were divided according to the degree of cervical maturation (shown by Bishop score) into two groups. In group I there were 60 women with unripe cervix (Bishop score < or = 5) and induction was performed by endocervical application of 0.5 mg Dinoprostone, and in group II there were 30 women with ripe cervix (Bishop score < or = 6) in which the induction was performed by intravenous application of Syntocinon (500 ml of 5% glycose with 10 iu of Syntocinon). After three hours the initially significantly different Bishop score values were equalized and it was possible to correlate the duration of the labor (Group I 545 min, Group II 338 min). The correlation of initial Bishop score values and the total duration of the labor was negative and high in both groups. All the neonates from both groups were in good condition and no serious complication was noted. Both methods of labor induction appear similarly effective and safe both for the fetus and the mother. PMID- 9863333 TI - [Relation between mitral orifice surface area and extent of hemodynamic disorder of the pulmonary circulation in patients with mitral stenosis]. AB - Mitral annulus and valves form the mitral orifice area with the size between 4.0 6.0 cm2. Every area which is smaller than this, represents mitral stenosis. As a consequence of mitral stenosis hemodynamic gradients occur over the mitral orifice with circulation disturbances below and above the stenotic mitral valve. The size of transmitral gradient is important in the evaluation of functional or/and structural changes in the blood vessels of pulmonary circulation. This investigation included 40 patients with mitral stenosis (or accompanying minimal mitral regurgitation). All patients underwent echocardiographic examination: area of the mitral orifice was determined and hemodynamic procedure with the left and right heart catheterization was performed. The following hemodynamic parameters were measured: mean capillary wedge pressure, left ventricular filling pressure, left ventricular mean diastolic pressure, mean pulmonary artery pressure. According to these parameters resistance in the pulmonary circulation was measured. The size of the mitral orifice was determined according to oximetry blood analyses and hemodynamic parameters. All patients were divided into 4 groups: minimal (2.5-4.0 cm2), mild (1.5-2.5 cm2), moderate (1.0-1.5 cm2) and severe mitral stenosis (1.0 cm2). The comparison of echocardiographic and hemodynamic parameters revealed a high and positive correlation between the area of mitral orifice. There was also a negative and moderate correlation between the values of stenotic mitral orifice area and total pulmonary resistance, i.e. in all patients with severe mitral stenosis there was an increased pulmonary arteriolar resistance. CONCLUSION: Noninvasive echocardiographic method is valid in the evaluation of stenotic mitral valve area. In the evaluation of hemodynamic parameters in the pulmonary circulation the index of arteriolar pulmonary systemic vascular resistance is very important. In all patients with the area of stenotic mitral orifice 1.0 cm2, there are functional or pathomorphologic changes in the pulmonary circulation of the blood vessel wall. PMID- 9863334 TI - [Correlation between the clinical and pathohistologic diagnosis in "small biopsies" of the lung]. AB - INTRODUCTION: During the last 20 years routine application of various methods of multiple "small biopsies" of the lungs such as forceps, transbronchial, trucut percutaneous and so on, has significantly increased the efficacy of diagnostics of bronchopulmonary and pleural diseases. Tissue samples, not bigger than 3-4 mm, in which diagnostic pathological changes are expected on the basis of previous clinical, radiological and bronchoscopic examinations, can be the basis for making a definite therapeutical decision only if a skillful surgeon has performed the biopsy by correct instruments and from the right place and sent it for histological analysis with other important clinical information. This study is a comment on quality, significance and possibilities of improving clinical pathological cooperation in this field of clinical pathology. MATERIAL AND METHODS: By correlation of clinical and histological diagnoses we analyzed the diagnostic efficiency of microscopic examinations of "small biopsies" of the respiratory tract in 319 patients (175 bronchial forceps biopsies, 31 transbronchial biopsies, 22 percutaneous needle pleural biopsies and 91 combined forceps and transbronchial biopsies) in whom biopsies were performed during 1996 in the Specialized Hospital for Lung Diseases Brezovik. RESULTS: Overall concordance between the clinical and histopathological diagnosis was 82.2%. In 99 cases (73.3%) out of 135 clinically "obvious" neoplasms, the histopathological examination confirmed existence of malignant tumor: squamous cell carcinoma in 80%, small cell carcinoma in 9.6% and adenocarcinoma in 5.6% of patients. In other patients it was not possible to perform a more precise classification. Endoscopic specimens of 29 patients (9.1%) were not representative. CONCLUSION: The level of diagnostic efficiency (73.3%) of definitive histopathological verification of bronchopulmonary lesions, which have been clinically diagnosed as malignancies, is rather high, but the increase of diagnostic efficiency requires application of more sophisticated histological diagnostic methods (immunohistochemical) and more frequent utilization of bioptic procedures which are more convenient for detection of peripheral pulmonary lesions (transbronchial and percutaneous fine needle aspiration biopsies of the lungs). PMID- 9863336 TI - [Aorto-enteric fistula as a rare manifestation of rupture of an abdominal aortic aneurysm--case report]. AB - Aortoenteric fistula represents a communication between aortic and duodenal lumen. It is a very rare complication of aortic aneurysmal disease as well as aortic prosthetic surgery. There are only few surgeons who have seen more than one case of aortoenteric fistula. At the Institute of Surgery one case of aortoenteric fistula was treated. It was well diagnosed at the Institute for Internal Medicine, and then presented to the vascular surgeon who operated immediately. The whole diagnostic and therapeutic procedure was well done, but stenosing carcinoma of ascendent colon was found during the operation. Resection were done leading to the dehiscence of bowel anastomosis and fatal outcome for the patient. This case is instructive in a diagnostic and surgical manner. PMID- 9863335 TI - [Herpes simplex and lymphocytic choriomeningitis viruses in infections of the central nervous system--clinical and cerebrospinal fluid characteristics]. AB - INTRODUCTION: A great number of various viruses are stated as the cause of acute infections and damages of the central nervous system. In most cases these are minor damages which exhibit as meningeal syndrome and a specific finding in the cerebrospinal fluid. According to the dominant location, central nervous system infections can take a form of meningitis, encephalitis or myelitis. Since the inflammatory process of the meninges can not be separated from the inflammatory process of the brain, we usually speak of meningoencephalitis. The etiological diagnosis of meningitis and encephalitis is established by isolating the virus from the cerebrospinal fluid and by finding the presence of the specific antibodies in the blood and in the cerebrospinal fluid. The most common causes of the viral meningitis are Enteroviruses, the Mumps virus, Arthropode borne viruses, the Herpes viruses, Adeno viruses and the Lymphocytic choriomeningitis virus. The aim of our study was to establish the correlation between the clinical features and immunological and cerebrospinal fluid changes and the degree of the damage to the blood-brain barrier during the infections of the central nervous system, caused by the Herpes Simplex virus and the Lymphocytic choriomeningitis virus. MATERIAL AND METHODS: From a group of 103 patients, who had been treated for viral meningitis and meningoencephalitis, a group of 27 patients with established specific viral etiology--Herpes Simplex virus and Lymphocytic choriomeningitis virus, had been taken into the account. Herpes Simplex infection had been proven by the complement binding reaction and the neutralisation test of the even samples of serum. The diagnosis of Lymphocytic choriomeningitis was confirmed by the immunofluorescence test of the pharynx swabs and cerebrospinal fluid. The clinical features, such as body temperature, encephalitic signs, and electroencephalographic findings had been followed and compared. RESULTS: Herpes Simplex infection had been found in 20 patients, Lymphocytic choriomeningitis had been proven in 7 patients. All the patients had increased body temperature. Only four of the patients exhibited encephalitic signs, all infected by the Herpes Simplex virus. Patients from the Herpes Simplex group showed various degrees of consciousness disturbances, ranging from somnolence to coma, while the Lymphocytic choriomeningitis patients exhibited none. Higher pleocytosis and protein level had been found in the Lymphocytic choriomeningitis group. DISCUSSION: Viral diseases of the central nervous system are the result of the direct damage of the brain and meninges by the virus and immunological processes. Herpes Simplex meningitis usually has a good prognosis. Lymphocytic choriomeningitis has longer course of the disease and exhibits more severe clinical features. CONCLUSION: In cases of the central nervous system infections, caused by Herpes Simplex virus or Lymphocytic choriomeningitis virus, the correlation between the severeness of clinical features and the degree of damage of the blood-brain barrier, the level of pleocytosis and the increase of the cerebrospinal fluid proteins had been established. PMID- 9863337 TI - [Gingival hyperplasia during treatment with nifedipine]. AB - INTRODUCTION: Nifedipine is a relatively new and increasingly used medication for treatment of all kinds of angina pectoris and arterial hypertension. The principal action of nifedipine is to inhibit the influx of extracellular calcium ions across the membranes of cardiac and vascular smooth muscle cells, without changing serum calcium concentration (1). One of the side effects of this drug is gingival hyperplasia which was first described in 1984 by Lederman (2). He noted that gingival hyperplasia is mostly marked on the labial gingiva of the upper and lower anterior teeth and appeared 1 to 2 months after nifedipine therapy began at a dose of 90 mg per day, and never occurs in edentulous areas. This paper reports gingival hyperplasia in a patient treated with nifedipine. CASE REPORT: A 73-year old man referred to the dental clinic for evaluation of gingival enlargement in the maxillar left quadrant which he had noticed about 3 months after starting the nifedipine therapy. Clinical examination showed gingival enlargement around the teeth 22, 23, 24 and 27, but with normal edentulous area (Fig. 1). The enlarged gingiva was red, smooth and shiny, with no pain on touch, and bled easily on probing. There were metal crowns with overhanging margins and pseudopockets--6 mm with dental plaque and calculus. Radiographs showed moderate alveolar bone resorption of horizontal type. The patient's maxillar left first premolar exhibited severe bone loss, resulting in a hopeless prognosis. This tooth was extracted under local anesthesia. The dental treatment included replacement of the ill-fitting metal crowns, scaling and root planning, and instructions on appropriate method for brushing teeth. 6 weeks later, the hyperplastic tissues were removed surgically. In a 1-year follow-up period, the patient was recalled at regular intervals for control, and no signs of recurrence of gingival hyperplasia have been observed in spite of continued nifedipine treatment. Biopsies taken from the gingivectomy specimens were handled with standard histological method. Histological examination showed a thick epithelium with parakeratosis and acanthosis, and irregular elongation of the rete peg. The underlying connective tissues contained dense fibers with inflammatory cell infiltrate mainly composed of plasma cells. DISCUSSION: The exact mechanism of action of nifedipine in causing gingival enlargement is unknown at present. There is also no answer to the question why gingival enlargement appears in some patients treated with nifedipine, but in others not. The clinical findings (3, 4, 5, 6, 7) and results of in vitro study (8) and this case report showed that hyperplastic change of gingiva started only in areas displaying signs of inflammation, but not in healthy and edentulous areas. These observations suggest that local factors and associated inflammation is essential for onset of the nifedipine-induced gingival hyperplasa. Moreover, it was suggested that duration of nifedipine therapy (more than 2 months) and drug dosage (90 mg per day) could be important (1,2,3). This case report also demonstrates that no signs of recurrence of hyperplasia were observed after elimination of the local inflammatory factors: extensive dental pluque control and surgical removal of the hyperplastic gingival tissue even though administration of nifedipine was continued. CONCLUSION: It was concluded that gingival enlargement occurs in patients with nifedipine therapy only in the areas where local inflammatory factors are present. PMID- 9863338 TI - [Shock]. AB - Shock is a consequence of disturbed circulation and decreased blood perfusion of tissues which, on the whole, cause functional and morphological impairments of organic systems. Occurrence of shock is not high, but it is significant due to bad prognosis and high mortality rate. CLASSIFICATION: This is one of the classifications of shock: cardiogenic, extracardiogenic-obstructive, oligentic and distributive shock. PATHOPHYSIOLOGIC MECHANISMS OF SHOCK: Regardless of the cause of shock, the clinical picture is dominated by hypotension mostly caused by decrease of minute volume (septic shock may be an exception--minute volume might be high). Due to hypotension a lot of compensatory mechanisms are activated and in the beginning showing the compensatory phase of disease and if nothing is done decompensatory and in the end irreversible phase of shock occurs with fatal outcome. The clinical picture depends on the etiology of shock, type of shock, but it also has certain specificities. PROGNOSIS: The prognosis depends on the clinical picture dominated by the following symptoms: hypotension, decreased diuresis, acidosis, consciousness disorders, tachypnea, peripheral cyanosis, cold and damp skin. MANAGEMENT: These patients are managed in intensive care units, where the following parameters are followed-up: arterial pressure, central venous pressure, minute heart volume, systemic vascular resistance, diuresis, continual follow-up of heart rate, breathing and consciousness. In treatment of shock it is necessary to: normalize the circulatory volume, establish electrolyte balance, regulate glycemia and acid-base status, compensate the volume, calm the patient and alleviate pain. PMID- 9863340 TI - Alternative therapy. PMID- 9863339 TI - [Development of orthopedics and founding of the Clinic for Orthopedic Surgery and Traumatology in Novi Sad]. AB - A study of the development of the orthopaedic surgery in Novi Sad from the antique times till 1980 when the Clinic for Orthopaedic. Surgery and Traumatology was established has been carried out. Development from Roman Times till the Second World War. The first material evidence of the surgical activities in this area originates from the epoch of the Roman Empire, when under the emperor Augustus (Augustus, 27 B.C.-14 A.D.), the northern border of the Empire was established and fortified on the river Danube. At that time on the place of the contemporary Petrovaradin Fortress, a small fortification with a permanent garrison, called Cusum, was built. As it was proved by excavation of the surgical instruments from the Roman time at the nearby Sremska Mitrovica (Sirmium) the surgery in the Roman legions was quite developed and it could be concluded that some forms of surgery and orthopaedics were practised in the antique Cusum as well. There is no reliable evidence about the health service in the area after the Roman era till the 1689 when Petrovaradin was finally sized by Austrian troops from the Turks, and when the first recorded hospital (Petrovaradin Military Hospital) in this region was founded. At that time on the northern shore of the river Danube a new settlement, developing later into the town of Novi Sad, was established. Surgery together with orthopaedics at that time were performed by the so-called "feldschers", and then by primitive, self-taught surgeons who were actually experienced barbers, while educated doctors started to practice surgery only in the 19th century when the first surgical department in Novi Sad was founded in the Great Town Hospital, and for years after that, the orthopaedics was practised by general surgeons. The foundation of the Anglo Yugoslav Children's Hospital for Tuberculosis of the Bones and Joints at Sremska Kamenica in 1934 by Dr. Katherine S. Macphail, a great benefactress of our people, was of great importance for the development of orthopaedics in this area as it was the first orthopaedic hospital in the country. The first small unit for orthopaedics, as a part of the Surgical Department of the Novi Sad State Hospital, was established in 1937 and led by a surgeon, Dr. Nikola Pravdica. Development from the Second World War till the 1980. After the Second World War the Main Provincial Hospital with two surgical departments was established in Novi Sad. At the so-called Second Surgical Department led by Dr. Nestor Teodorovic a small unit for traumatology and orthopaedics was founded in 1946, which functioned as such till 1953. The first properly trained orthopaedic surgeon was Dr. Stojan Letic who passed the specialty exam in 1953 and was attached to the Second Surgical Department of the Main Provincial Hospital. The first independent Orthopaedic and Traumatologic Department, led by an experienced orthopaedic surgeon, Dr. Vladimir Razenj, was established in 1956. From 1959 till 1963 the department was led by Prim. Dr. Dobrivoje Gradistanac. In 1963 this department together with the former "English Hospital" at Sremska Kamenica were incorporated into the so-called Clinic for Surgical Diseases and divided in the Orthopaedic (led by Prof. Milenko Dosen) and separated Traumatologic (led by Prim. Stojan Letic) departments which existed as such till 1975 when they were integrated again into a single unit which evolved into the Clinic for Orthopaedic Surgery and Traumatology in 1980. PMID- 9863341 TI - Sports medicine across Missouri: interviews with Dr. Paul Meyer and Dr. Stanley London. Interview by Thomas D. Eppright, Shane Bradley, Maureen Alwood. AB - Paul Meyer and Stanley London are two Missourians who have led the way for many in the field of Sports Medicine. The careers of many professional athletes have been extended due to the clinical expertise of these exemplary physicians who both began their medical careers in the 1940s. Through this interview, they offer a historical perspective of Sports Medicine and baseball. They also share their life experiences, as well as their formulas for successful careers and long lasting marriages. PMID- 9863342 TI - Prevalence changes in modifiable cardiovascular disease risk factors in three Missouri regions, 1990-1996. AB - OBJECTIVES: To determine the prevalence and prevalence trend of modifiable cardiovascular disease (CVD) risk factors among African Americans and whites/others from 1990-1996. We also examined differential changes between African Americans and whites/others during the same time period. METHODS: This study used data from two special Behavioral Risk Factor Surveillance System (BRFSS) based surveys in 1990 (N = 3,000+) and one in 1996 (N = 2,095) that targeted two metropolitan and one rural region in Missouri with substantial minority populations. Risk factors included physical inactivity, obesity, hypertension, unmonitored cholesterol and smoking. The percent change in prevalence estimates and corresponding confidence intervals between survey years were calculated for each of the above risk factors. RESULTS: When compared with Missouri BRFSS data, overall prevalence of smoking, obesity, hypertension and unmonitored cholesterol was higher in the three-region study population than the state as a whole. African-American males did not experience any statistically significant reductions in CVD risk factor prevalence rates between 1990 and 1996, while the increase in obesity was driven mostly by the African-American female subgroup. The least amount of reduction in CVD risk factors was seen in individuals aged 55 or older, with a high school education or less and/or without health insurance. CONCLUSIONS: Individuals of African-American ethnicity, aged 55 or older, with a high school education or less and/or without health insurance need to be the focus of future public health initiatives designed to reduce the prevalence of CVD risk factors. PMID- 9863343 TI - Trends in breast cancer screening in Missouri from 1987 to 1995, and predictions for the years 2000 and 2010. AB - BACKGROUND: Breast cancer is a major cause of morbidity and mortality in the United States (U.S.) and Missouri. In 1992, 3,915 new breast cancer cases were diagnosed and in 1995, 1,006 deaths from breast cancer were reported in Missouri. Although breast cancer incidence has increased in Missouri in the past 20 years, there are indications that early detection has also increased during the same period. Knowledge about which segments of the population have experienced the greatest increase in mammography screening rates helps in planning and implementation of breast cancer control programs at the state level. OBJECTIVES: Examine the prevalence and trends of lifetime mammography and 2-year mammography compliance in Missouri by age, race, and education from 1987 to 1995 and make predictions for the years 2000 and 2010. METHODS: We used data from the Missouri Behavioral Risk Factor Surveillance System (BRFSS), 1987 to 1995, to estimate the prevalence of ever having had a mammogram and compliance with mammography screening guidelines within two years by race, age, and education status among Missouri women over age 18. Using linear models, we regressed breast cancer screening prevalence estimates on time to obtain trends and predictions. RESULTS: Overall, African-American women were more likely to have had a lifetime mammogram than white women. However, we found a steady increase in the prevalence of ever having had a mammogram for all groups of women defined by age and education status, except among African Americans. Increase in the prevalence of ever having had a mammogram was much higher in women age 50 and older and slightly higher among women with a high school education or less. The average prevalence of 2 year mammography screening compliance was about 60% for all groups, a rate which did not significantly change between 1987 and 1995. By the year 2000, white women will have mammography rates equal to or higher than African-American women, and the majority of all women age 50 and older (98.3% to 100%) will have had a lifetime mammogram. CONCLUSION: Missouri target populations are predicted to attain Year 2000 National Health Objectives concerning lifetime mammography. Current efforts should be continued in order to maintain levels of mammography, particularly among African-American women. PMID- 9863344 TI - Endoscopic carpal tunnel release: its time has come. PMID- 9863345 TI - [Ethical aspects of social changes in medicine after 1989]. PMID- 9863346 TI - [Interdisciplinary cooperation in thyroid diseases--views of the surgeon]. PMID- 9863347 TI - [Thyroid gland surgery at the 1st Surgical Clinic of the 1st Medical School of Charles University and General Medical School Hospital in Prague--review of problems]. AB - The authors analyze a group of 100 patients indicated for goitrectomy during the period from 1995-1996. The most frequent indication for surgery was an eufunctional polynodular goitre. The group comprised 86 women and 14 men. Their mean age was 47 years, bilateral affections predominated. The most frequent type of operation was total thyroidectomy. Carcinoma was found in three of the operated patients. The mean period of hospitalisation was 6 days. Postoperative complications were more frequent after total goitrectomies, and with the extent of the operation the number of possible complications increased. The most serious complication in the authors opinion was a lesion of the parathyroid glands, observed in 5% of the patients. Permanent unilateral lesions of the recurrent nerve were recorded in 2% of the operated patients. There was no lethal postoperative complication. The largest operated goitre weighed 625 g (photograph 1). The authors conclude that it cannot be stated that the operation of larger goitres is associated with the highest incidence of postoperative complications. The opposite is rather the case. PMID- 9863348 TI - [Minimally invasive treatment of traumatic cerebral hematoma. Further experience with stereotaxic evacuation]. AB - The authors summarise their experience with 16 post-traumatic cerebral haematomas which they evacuated using stereotactic technique. Based on the achieved results they reached the conclusion that stereotactically aimed treatment of haematomas is an advance, for the patient's benefit. The evacuation cannula is safely inserted to its target from a transcutaneous drill hole of the skull and thus the patient is spared the surgical trauma associated with craniotomy and further brain damage. If necessary, the focus of haemorrhage can be reached again by puncture via the channel in the bone which navigates the cannula. Surgery under local anaesthesia is rapid and the results of promising. Extension of the new method depends, however, on the availability of simple stereotactic equipment which starts to play a part in the surgery of traumatic haematomas. PMID- 9863349 TI - [Combined surgical treatment of portal hypertension]. AB - The bleeding from esophageal varices is at 85-90% cases stopped by conservative (i.e. non surgical) approach. The method of choice is endoscopic sclerotisation or ligation. Less often is necessary to perform TIPS. The rest of cases is necessary to resolve by one of surgical techniques of hemorrhage control- devascularisation operation or perform emmergently porto-systemic shunt. The authors stress the importance of devascularisation operations for theirs simplicity. On the case report they illustrate the advantage of combination of both surgical techniques. PMID- 9863351 TI - [Laparoscopic resection of a mesenteric cyst]. AB - The authors report a case of an unusual abdominal tumor-mesenteric cyst. When it is found active surgical approach is recommended. Laparoscopic resection of the cyst is considered as the method of choice. PMID- 9863350 TI - [Laparoscopic gastrostomy: report on 20 cases]. AB - At the surgical clinic of the Faculty Hospital in Ostrava-Poruba since November 1993 till June 1998 20 laparoscopic gastrostomies were performed. The main indication was to ensure enteral nutrition in patients where, due to obstruction of the oesophagus, it was impossible to implement percutaneous puncture gastrostomy. The surgical procedure lasted on average 37 minutes and was completed successfully in all patients. Three patients died within thirty days. This procedure made it possible that eighty percent of the patients could be discharged from hospital and go home. PMID- 9863352 TI - [Use of the Miller-Abbott tube in treatment of early postoperative ileus]. AB - The authors present an account on the use of an intraluminal intestinal stent to treat early postoperative ileus. They demonstrate on a case-history the possibility to combine the tube inserted via jejunostomy and with laparostomy. In the discussion they deal also with the historical development of intubation of the small intestine and support of controlled adhesion formation. PMID- 9863353 TI - [Dr. Frantisek Burian and burn care in Europe]. PMID- 9863354 TI - [Surgical treatment of a complication of Crohn's disease of the duodenum]. AB - In 1995-1997 at the First Surgical Clinic, Medical Faculty, Palacky University and Faculty Hospital Olomouc three patients were treated on account of obstruction of the pyloroduodenal region caused by Crohn's disease. The condition was treated by anastomosis, possibly with vagotomy, depending on the site of stenosis. Clinical, laboratory and endoscopic control examinations after a 6 month to 3-year interval revealed good results of this approach. PMID- 9863355 TI - [Sentinel lymphadenectomy in malignant cutaneous melanoma--short-term results and prognostic significance]. AB - The predictive value of sentinel node status in skin melanoma was studied. Between August, 2, 1994 and February, 20, 1998, in Surgical Department of Masaryk Memorial Cancer Institute, Brno, 166 lymphatic regions were explored in 153 patients with 154 melanomas. We were not able to visit any other institute performing this procedure, so our beginning failure rate has been quite high, approx. 40% declining to final less than 3%. Lymphatic mapping was performed with the use of Patentblau V dye (BYK, Germany), injected preoperatively intradermally to the tumour or into the scar after nonradical removing of melanoma. Lyphoscintigraphy has been used in cases of dubious lymphatic spread, especially in trunk melanomas. Hand held gamma probe could not be used for unacceptable cost. From 143 identified sentinel nodes were 26 positive in 24 patients. From these patients 10 relapsed, with 6 locoregional and 4 distant recurrences. 2 Patients died. In 108 patient with negative sentinel node 3 recurrences occurred, one nodal, and two local, no distant metastases occurred till now. The incidence of positive sentinel nodes was growing with Breslow thickness. Extremely poor prognosis can be expected in patients with Breslow > 4.0 mm and positive sentinel node. PMID- 9863356 TI - [Disseminated intravascular coagulation and hyperfibrinolysis in dogs with metastasizing mammary carcinoma]. AB - The alterations of the haemostatic system (platelet count, activated partial thromboplastin time [APTT], thromboplastin time [standard test, modified test], thrombin time, fibrinogen concentration, activity of the coagulation factors II, V, VII, X, VIII:C, IX, XI, XII, of prekallikrein, high molecular weight kininogen, antithrombin III, protein C, plasminogen and alpha 2-plasmin inhibitor, concentration of soluble fibrin and fibrin(ogen) degradation products [FDP], resonance thrombogram) were described in seven dogs with haemorrhagic diathesis in consequence of an infiltrative, growing mammary carcinoma with multifocal invasion of lymphatic and blood vessels. In most of the cases metastases in different organs could be demonstrated. In every case a serious stage of disseminated intravascular coagulation and hyperfibrinolysis was existent. This was indicated by the distinctly increased concentration (p < 0.0001) of soluble fibrin (27.7 [16.0-79.2] micrograms/ml, median [minimum maximum], reference range [RR.]: < 9.4 micrograms/ml) and FDP (340 [50-860] micrograms/ml, RR.: < 18 micrograms/ml) as well as a diminished plasma level of nearly all components of the coagulation and fibrinolytic system concerning especially the concentration of fibrinogen (0.16 [0.01-0.46] g/l, RR.: 1.17-3.09 g/l), the activity of factors V (30 [21-40]%, RR.: 75-158%) and VIII:C (9 [4 16]%, RR.: 72-136%) as well as the activity of protein C (8 [3-13]%, RR.: 68 139%) (each: p < 0.0001). PMID- 9863357 TI - [Evaluation of the portable blood analyser i-STAT]. AB - The purpose of this study was to compare the results of horse blood examinations on a portable blood analysis system, i-STAT SDI, with conventional laboratory equipment and to verify the accuracy and precision. The precision within run was high and the coefficients of variation ranged from 0 to 3.85%. The comparative analysis of patient samples with routine methods showed a very strong positive correlation with correlation coefficients above 0.96 for blood gases, pH, glucose, urea, hemoglobin, hematocrit, sodium and potassium, with the exception of chloride where it was 0.867. The i-STAT system provides a quick and accurate stable-side evaluation of laboratory data in critically ill horses and emergency situations. PMID- 9863358 TI - [Continuous serologic study and sanitation inspection of Sarcoptes scabiei var. suis infection: preliminary results]. AB - Pig mange was investigated serologically with ELISA using Chekit Sarcoptest. The test consists of a homogenized antigen of Sarcoptes scabiei var. vulpes. A total of 356 sows sera samples were collected from 16 mange free breeding units and 19 units with clinical manifestation of mange infestation. The specificity of the test was 99.34% and the mean sensitivity value was 62%. This test will allow in the future not only the rapid and certain diagnosis of pig mange but also the screening of the pig breeding units after eradication of the disease. PMID- 9863360 TI - [Importance of the number of trinucleotide repeat expansions in the clinical manifestations of Huntington's chorea]. AB - INTRODUCTION: In 1993 the gene responsible for Huntington's disease (IT15) was isolated [5]. It was mapped to the tip of the short arm of chromosome 4 and within its coding sequence, near the 5' end, it contained a certain number of trinicleotide (CAG)n (cytosine-adenine-guanine) repeats (Figure 1). This gene codes for a protein (348 kd) called "huntington" that is widely expressed, and its sequence is not related to any protein [6]. The normal range of (CAG)n repeat numbers within IT15 was reported to be between 6 and 37 [6]. Mutation responsible for Huntington's disease implied expansion of (CAG)n repeats: in patients with Huntington's disease the pathologic range was determined to be between 35 and 121 repeats [7-10]. PATIENTS AND METHODS: In this study we correlated the age at onset, rate of progression and initial symptoms of Huntington's disease with the number of trinucleotide (CAG)n repeats in IT15. DNA was isolated from peripheral blood leukocytes of patients fulfilling clinical criteria for definite and probable Huntington's disease [2]. Genetic verification of Huntington's disease was made by the previously described and modified PRC (polymerase chain reaction) technique [17, 18]. In our laboratory a gene with 40 or more repeats was considered as a marker of Huntington's disease. RESULTS: The study comprised 26 patients (11 women and 15 men). At the onset of Huntington's disease they were between 19 and 66 years old (36.6 12.8 years), with the duration of the disease between 1 and 15 years (5.8 4.3 years). The number of (CAG)n, repeats in IT15 ranged between 40 and 95 (49.9 14.1). The negative correlation between the (CAG)n, count in the expanded allele and the age at onset of the disease has been confirmed. Regression analysis showed the correlation coefficient of -0.54 (p = 0.012). The effect of trinucleotide (CAG)n, repeats on the initial clinical manifestations and rate of progression of Huntington's disease is only one of the growing group of "CAG-repeat" disorders that also include entities such as spinocerebellar ataxia-type 1 and 3, spinobulbar muscular atrophy and dentato rubo-pallidoluysian atrophy [6]. PMID- 9863362 TI - [Interleukin 4, total and allergen-specific immunoglobulin E antibodies in the blood of individuals with an atopic constitution]. AB - INTRODUCTION: The recently published data suggest that atopic patients have an enhanced ability to produce IL-4, even in response to antigens other than common environmental allergens or helminthic components. This aberrant IL-4 production by Th cells may be one of the immune alterations encoded by nonMHC genes in the control of basal IgE level [1-7]. The existence of atopen-CD4+ T cell clones that have diverse repertoires might be relevant for aetiopathogenesis of atopic diseases [8-10]. Being subjected continuously to the environmental antigens (allergens), the immune system, especially T cells, is permanently activated and stimulated in response to that challenge. Due to that, substantial amount of diverse cytokines (and their soluble receptors) could be found in the serum. Accordingly, it could be expected to find raised serum IL-4 concentration in atopic persons. In this study we investigated concentrations of IL-4, total IgE and allergen-specific IgE in sera of atopic persons susceptible to pollen allergens. The main goal was to estimate whether the immunologic profile of atopics is defined by increased serum concentration of IL-4 apart from total and allergen-specific serum IgE. METHODS: Patients. We selected 9 atopic patients in the range of 19-35 years, susceptible to grass or weed pollens and suffering from allergic rhinoconjuctivitis. There were 6 females and 3 males (mean age 31.2 years). Atopic allergy was proven by clinical history, skin prick test and by means of in vitro test (positive serum allergen-specific IgE antibodies, RAST/radioallergosorbent tests [11]. As controls we randomly selected B blood donors (6 females, 2 males, mean age 31.3). Study design. The study was carried out from August till September 1995. None of selected patients had previously accomplished allergen-specific immunotherapy nor had been medicated by corticosteroids (topic or systemic) [12-15]. Selected persons were not allowed to take medications such as antihistamines, beta-2 agonists or tricyclic antidepressants at least 10 days prior to the study. We performed in vitro tests for determining concentrations of IL-4, total and pollen-specific serum IgE. Sera were sampled in the morning and were stored frozen at -20 degrees C until analysis. Determination of IL-4, total and allergen specific IgE in serum. Interleukin-4 measurement was carried out in blind fashion with an ELISA kit (Intertest-4 ELISA, Genzyme, Cambridge) according to the manufacturer's instructions. A reference curve was obtained by plotting the IL-4 concentration of several standard dilutions versus an absorbency. The determination limit was 0.045 pg/ml. For determining total serum IgE we used commercially available enzyme immunoassay (EIA Phadezym IgE PRIST, Pharmacia, Uppsala). Normal values was up to 120 IU/L (international unit per ml). Allergen specific IgE was determined by semiquantitative EIA method (RAST Phadezym, Pharmacia, Uppsala) and expressed in Phadebas RAST Unit (PRU) according to standard curve done by manufacturer. Study was carried out after approval of the Ethic Committee of our Hospital and with the obtained patients consents. For statistical analysis we used nonparametric tests (U-test and the Spearman rank correlation). For all comparisons, statistical significance was considered to be present if p < 0.05. RESULTS: Among selected patients, 4 persons had high concentration of serum IgE antibodies (RAST class 4) against grass pollens, two persons had IgE-specific (RAST, class 4) to weed pollens and 2 patients had IgE antibodies (RAST class 4) against weed and grass pollens simultaneously. RAST of class 3 was registered in the serum of one person. Registered serum total IgE, allergen-specifc IgE and IL 4 are shown in Table 1. Serum IL-4 was significantly higher in atopics (U-test, SR 43.53, p.05) as well as total IgE (ER 493, p.05). In atopics, IL-4 was not in correlation with either total or allergen-specific IgE (IL-4 versus total IgE, p = +0.190; PMID- 9863361 TI - [Doppler sonography diagnosis of restenosis after percutaneous transluminal angioplasty: sensitivity and specificity of the pedal-brachial index in relation to changes in absolute arterial blood pressure]. AB - INTRODUCTION: Percutaneous transluminal angioplasty (PTA) is accepted for the treatment of patients with severe, disabling claudication who fail conservative management and also for patients with limb threatening ischaemia [1-5, 18, 20]. The development of neointimal hyperplasia (predominantly during the first 6-12 months after PTA), and the progression of the underlying atherosclerotic disease (thereafter), are the reasons of restenosis and reocclusion [1-4, 6]. More than 50% of occurring restenoses are primarily oligo/asymptomatic [1-4, 6-10, 25, 26]. Follow-up visits are aimed at detecting significant restenoses, before reocclusion occurs, so that timely reintervention is possible. In asymptomatic patients in whom reintervention is not necessary, repeated angiography is not justifiable. Non-invasive alternatives include Duplex scanning or the assessment of simple peripheral haemodynamic variables such as ankle systolic pressures and the ankle/brachial pressure index (ABI). The aim of this study was: (1) To determine the sensitivity and specificity of post PTA changes in the ABI, and changes in the absolute ankle pressure to detect restenoses after femoropopliteal PTA (as a gold standard, Duplex scanning, with its reported high sensitivity, specificity and accuracy for detecting restenosis was chosen [9, 25]. (2) For both methods, to evaluate the criteria (minimal magnitude of change-cut off points) necessary to detect restenosis with reasonable reliability. (3) To compare the diagnostic value of changes in ABI to changes in absolute ankle pressure, and to determine the method of preference for detection of post-PTA restenosis. MATERIAL AND METHODS: The study included 171 consecutive patients with peripheral arterial occlusive disease, Fontaine stage II or stage III, selected for femoro-popliteal PTA. All pts presented with single or multiple arterial stenoses or occlusions not exceeding 10 cm of length within the femoro popliteal segment. At least one lower leg artery had to be patent. Only pts in whom PTA was successful (maximum residual lesion within the dilated segment showed < or = 30% diameter reduction (DR)) were accepted for a 12-month follow up. After PTA all patients were prescribed a platelet aggregation inhibitor. The investigational scheme included the following procedures: 1. Duplex scanning of the entire lower leg vascular tree before PTA, within 7 days after PTA as well as at 4 weeks and at 12 months. 2. Scoring of Duplex results using the Bollinger angiography score system [11] which distinguishes the pelvic segment, a proximal and a distal superficial femoral segment, the popliteal artery including the popliteal trifurcation and the proximal 1/3 of the lower leg arteries. The score system allows the differentiation of single and multiple plaque (< or = 25% DR), single or multiple stenoses (< or = 50% and < or = 50% DR) involving less or more than 50% of the segment under investigation as well as short (< 50% of segment length) and long (> or = 50% of segment length) vascular occlusions. 3. The assessment of the resting ankle systolic pressures (dorsalis pedis artery, posterior tibial artery) of the reference leg, pre PTA, at 4 weeks, and 3, 6, 9 and 12 months using a 8 Mhz CW Doppler device (Parks 908) with the patient in the supine position and after a resting period of > or = 30 minutes. 4. The calculation of the ankle/brachial pressure index (ABI) as the ratio of the higher systolic pressure value from either the posterior tibial or the dorsalis pedis artery divided by the higher value of the two (bilateral) systolic brachial pressures. 5. A full physical examination including a pulse status, vascular auscultation as well as the assessment of the clinical symptomatology pre PTA, at 4 weeks as well as at 6 and 12 months. The following findings were suggestive of restenosis/reocclusion of the dilated segment. 1. Reoccurrence of a stenosis < or = 50% DR within the dilated segment and/or its inflow or outflo PMID- 9863363 TI - [Elevated Epstein-Barr virus antibodies in patients with Hodgkin's disease]. AB - INTRODUCTION: Some studies of Hodgkin's disease support the concept that the disease may be due, directly or indirectly, to a common infectious agent, an infection acquired in late rather than early childhood being associated with an increased risk of oncogenesis. The Epstein-Barr virus is the leading candidate as one of these causative agents, because of its oncogenic properties in the laboratory and its association with two human malignancies, African Burkitt lymphoma and nasopharyngeal carcinoma. METHODS: We studied 49 patients with Hodgkin's disease. The patients were grouped according to the histologic type, clinical symptoms and immune state. The control group of 140 subjects was selected among healthy people. The antibodies against viral capsid antigen were measured with immunoperoxidase test, antibodies to nuclear antigen with anticomplement immunofluorescent test, and antibodies to early antigen with immunofluorescence method. RESULT: We found that all patients had antibodies against viral capsid antigen and nuclear antigen. We found an elevated titers for viral capsid antigen in 69% of patients, early antigen in 41% and nuclear antigen in 55% of patients. The incidence of elevated titers (> or = 320 for viral capsid antigen and > or = 40 for early antigen) and nuclear antigen was higher in patients than in control group; the differences were statistically significant. Also, the geometric mean of early viral capsid and of nuclear antibody titers in patients were significantly higher than that in control group (p < 0.01). DISCUSSION: The possible role of Epstein-Barr virus in Hodgkin's disease has been long investigated in epidemiological studies and the finding of Epstein-Barr antibody has been currently supported by the detection of genomic Epstein-Barr virus in diagnostic cells in some cases of Hodgkin's disease. Hodgkin's disease varies in prevalence, morphologic findings and association with Epstein-Barr virus in different parts of the world. Hodgkin's disease in Yugoslavia and its relationship to Epstein-Barr virus has not yet been studied. In conclusion, our results indicate that elevated titers of antibody against antigens associated with Epstein-Barr virus exist in patients with Hodgkin's disease. Whether this finding reflects a direct role of Epstein-Barr virus or whether this activity is a marker of the effect of a more fundamental factor, is not yet clear. PMID- 9863364 TI - [Physical activity among school children]. AB - The aim of this paper was to assess pupil's habits concerning physical activity and to identify factors related to these habits. METHOD: The data in this analysis were taken from the baseline survey of "Healthy School Project". A total of 215 pupils from four schools completed self administered questionnaires (response rate 91.49%). The children obtained questionnaires for their parents and returned them completed the next day. A total of 201 parents completed the questionnaire (response rate 85.53%). The questionnaire covered different health related topics including physical activity. In this study we analyzed questions concerning the frequency of out school physical activity, attendance in physical education classes and attitudes towards physical activity. Data were analyzed using SPSS/PC. Gender differences and grade differences were assessed with chi square calculations. To assess the contribution of predictor variables, stepwise multiple regression analyses were computed, for the total sample and for the fifth and the eighth grades separately. RESULTS: The majority of pupils, more than 60 percent, reported involving in leisure time physical activity a few times a week or even every day. Chi-square showed no statistical gender difference in the fifth grade in reports on leisure time physical activity, but in the eighth grade boys reported significantly more regular physical exercise. Pupils reported a high attendance in physical education classes (99.1% in the fifth and 82.4% in the eighth grades). A large majority of pupils thought that they would be involved in physical activity in the future (84.2% in the fifth and 77.4% in the eighth grades). Multiple regression analyses showed that for the present physical activity of pupils the regular parent's physical activity was the strongest predictor. When done separately for the fifth and the eighth grades multiple regressions analyses showed that only for younger children the parents' physical activity was also the predictor of theirs. Multiple regressions analyses showed that for the attitude towards future physical activity the strongest predictor was the frequency of the present physical exercise, and it was so far both the fifth and the eighth grades. For pupils in the fifth grade parents' physical activity was also the predictor but it was not for the pupils in the eighth grade. DISCUSSION: A positive finding was that the majority of pupils reported regular physical exercise and intention to continue with physical activity in the future. In the fifth grade there was no significant difference in leisure time physical activity but in the eighth grade girls reported less leisure time physical activity, and this might be the result of traditional opinion that physical activity and physical fitness are very important for boys but not for girls [15, 16]. Multiple regression analyses also showed that physical activity was related to regular parent's physical activity for younger children, and also the parents' physical activity was the predictor of intention for involving in physical activity in the future; this means that parents' behaviour is the model for their children. PMID- 9863365 TI - [Fractures of the metatarsal bones caused by fatigue]. AB - The term stress (marching) fracture relates to a fracture provoked without acute trauma, on healthy bone and as a consequence of repeated, long-term, rhythmic load [4, 5]. Stress fractures are mostly observed in young people (12-25 years of age) [15]. The course of this phenomenon we found by medical examination; this means that it is found either by detection or spontaneous ambulance reports. The aim of this study is to point at a possible appearance of these fractures at young soldiers and sportsmen. MATERIAL AND METHOD: During the routine ambulance practice, and by the prospective study on 29 subjects, the study was performed by using oral inquiry [9]. The studied individuals were soldiers who were complaining of pain, swellings and redness of the feet. Except clinical orthopedic examination, additional laboratory analysis was made. RESULTS: The results are shown in tables. Table 1 shows that stress fractures of metatarsus bones usually appear during the period of first thirty days of the military service, i.e. during the days of the hardest exertion. The most frequent training absence, which is caused by stress fractures, is during this period of time (Table 2). Of all military duties, marching had the biggest influence on appearance of this disease (Table 3). DISCUSSION: On the basis of available literature it is evident that there are factors which might have the influence on the occurrence of the disease (hypovitaminosis, foot-wear, foot deformation, body weight, muscle fatigue, etc.). Stress fractures are mentioned in connection with some diseases or special clinical deformations such as rheumatoid arthritis, chronic bronchitis, arthrosis deformities, after arthroplastics, Mb. Bechterew, etc. Consequently, stress fractures do not appear only in a healthy bone, as it is defined [4, 5]. In the clinical practice it was established that the diagnosis of this pathology had, in a few cases, the same clinical picture. On the basis of our experience, by observing, we have concluded that stress fractures occur on healthy or changed bones as the result of physical fatigue of the material, and repeated movements. It is still unclear why these changes are unilateral. CONCLUSION: a) Stress fractures are frequently appearing at young soldiers. b) Most frequently they appear in beginning of hard and exhaustible training. PMID- 9863366 TI - [Hormones of the posterior region of the hypophyseal gland]. AB - Arginine vasopressin is the main hormone involved in the regulation of body fluid osmolality. The hormone is released by the posterior pituitary whenever water deprivation causes an increased plasma osmolality or whenever the cardiovascular system is challenged by hypovolaemia and/or hypotension. The main site of action of this hormone is the renal collecting duct, but vasopressin is also a potent vasopressor and neurotransmitter, it has a role in the secretion of corticotrophin, in the regulation of the cardiovascular system, temperature and other visceral functions. Vasopressin also promotes the release of coagulation factors by vascular endothelium and increases platelet aggregability. In addition to its classical contractile effect on uterine myometrial and mammary glandular myoepithelial cells, oxytocin acts as neurotransmitter, stimulates endometrial prostaglandin production, pituitary prolactin secretion, luteolysis, sperm transport and natriuresis, and may play a role in immune function. Sensorial stimuli arising from the cervix and vagina as well as stimulation of the breast can induce secretion of oxytocin from the posterior pituitary. There are many vasopressin and oxytocin analogues (agonists and antagonists) that are synthetized with the goal of increasing duration of action and selectivity for the receptor subtypes, while non-peptide antagonists are orally active. The oxytocin and the vasopressin V1a, V1b and V2 receptors have recently been cloned and shown to form a sub-family within the large superfamily of G-protein-linked receptors. Renal V2 receptors mediate vasopressin-induced water reabsorption via induction of intracellular cAMP production in collecting duct cells. Most remaining actions of vasopressin on blood vessel constriction, liver glycogenolysis, platelet adhesion, adrenal angiotensin II secretion and certain brain functions are mediated via V1a-type receptors that are coupled to a Gq/11 protein. V1 receptor activation leads to stimulation of phospholipases C, D and A2, and an increase in intracellular calcium. Vasopressin stimulates pituitary corticotrophin release via a third vasopressin receptor type (V1b) which is present in corticotrophs. Oxytocin induces myometrial contraction, endometrial prostaglandin F2a production, mammary gland milk ejection, renal natriuresis and specific sexual, affilitative and maternal behaviours via oxytocin receptors which are also coupled to a G1/11 protein. Although only one oxytocin receptor type has been cloned so far, recent binding studies indicate that uterine endometrial oxytocin receptors may constitute a distinct receptor subtype. Expression of oxytocin receptors have relevant up- and down-regulation by oestrogens and progesterone. PMID- 9863367 TI - [Diagnosis of Lyme neuroborreliosis]. AB - Lyme neuroborreliosis is a disease of the central and/or peripheral nervous system caused by spirochete Borrelia burgdorferi. The diagnosis of Lyme neuroborreliosis is based on the clinical picture, differential diagnosis, serological tests with two-step approach and confirmation of positive results, monitoring of antibody titers and effects of adequate therapy. In failure revision of diagnosis is necessary, and if the diagnosis of Lyme neuroborreliosis is correct the effects of the therapy should patiently be expected. Relapse or reinfection are confirmed by new raise in titer of specific antibodies, new symptomatology and new antigens. PMID- 9863368 TI - [Isolated systolic arterial hypertension in the elderly]. AB - Isolated systolic hypertension is a common disorder in the elderly carrying a high risk of stroke and cardiovascular disease. Isolated systolic hypertension is usually defined as a systolic blood pressure greater than or equal to 160 mmHg and diastolic blood pressure less than 95 mmHg. The arterial stiffening is the principal cause of increasing systolic pressure in advanced age. It is due to degeneration of the arterial wall and is associated with progressive arterial dilatation. Hypertension in elderly patients is also characterized by increase of peripheral vascular resistance. Due to the wide variability of blood pressure usually seen in old persons, the isolated systolic hypertension is not easy to recognize and final diagnosis requires a long period of observation. The ambulatory blood pressure monitoring proved to be helpful in distinguishing patients with true isolated systolic hypertension from subjects with exaggerated alarm reaction to the pressure measurement. Although the increased risk of cardiovascular and cerebrovascular mortality is well established for isolated systolic hypertension, there has been much debate whether available antihypertensive treatment can prevent or delay cardiovascular and cerebrovascular complications in this condition. During the last year several large new trials have been published, the so-called STOP-Hypertension, SHEP and MRC trials. All studies have demonstrated that the treatment of isolated systolic hypertension with diuretics or/and beta blockers (frequently used in combination) resulted in a significant reduction in the incidence of stroke and major cardiovascular events. New antihypertensive agents such as calcium channel blockers and angiotensin-converting enzyme inhibitors have also been shown to effectively lower systolic blood pressure in the elderly but the effects on long term morbidity and mortality are still unknown. PMID- 9863369 TI - [The role of eosinophilic leukocytes in allergic inflammation]. AB - Eosinophilic leukocytes are tissue cells of granulocytic structure and secretory nature. They are produced in the bone marrow and transported to the targeted tissue via the blood where they are present in concentrations hundred times higher than in peripheral circulation. Eosinophilic leukocytes are the essential effector of allergic inflammation, which is a pathophysiological basis of allergic diseases. These diseases are characterized by disturbed distribution of eosinophilic leukocytes, i.e., peripheral eosinophilia and/or infiltration of the affected organs. Migration of these cells from the peripheral circulation into the targeted tissues, i.e., affected with the allergic inflammation, is influenced by helper T2 cells-dependent cytokines, and other mediators of inflammation. Subsequent to their activation, eosinophilic leukocytes release numerous made and newly produced mediators of inflammation and also present antigens which define their effector function in allergic inflammation. In this way, eosinophilic leukocytes participate in numerous pathological and pathophysiological disorders characteristic of allergic diseases which clearly confirm the active role of these cells in their production. PMID- 9863371 TI - [Pseudoaneurysm of the gluteal artery: 2 case reports]. AB - INTRODUCTION: Gluteal artery pseudoaneurysms are very rare [1]. They mostly occur after gunshot and stub wounds [2]. However, gluteal artery pseudoaneurysms can be caused by pelvic fracture [1]. Also, they can be isolated or associated with trauma of the pelvic and abdominal viscera [3]. The authors present two cases of gluteal artery pseudoaneurysms. Case 1. A 30-year-old man was treated for large swelling of the left buttock. One month previously he manifested a gunshot wound in the gluteal region. He also had symptoms of lumboischialgia with peroneal nerve paresis. The physical examination revealed a large pulsatile mass over the left buttock with an associated overlying bruit. Selective angiography of the internal iliac artery (Figure 1) revealed a large inferior gluteal artery pseudoaneurysm that caused dislocation of both external and internal iliac arteries. The patient was operated under epidural anaesthesia by the combined abdominal (extraperitoneal) and gluteal approach. By extraperitoneal approach the internal iliac artery was identified and ligated. After the closure of the wound, the patient was placed on the abdomen, and pseudoaneurysm was opened by an incision made between gluteus maximus and medius muscles. After evacuation of the parietal thrombus and pseudocapsule resection, nutrient vessels were ligated. The postoperative recovery was good, and the patient was free of neurologic symptoms two days after the operation. The late result (after 4 years) is also good. Case 2. A-53-year-old man was treated for small haematoma pulsans (Figure 2) in the right buttock. Fifteen days previously he was treated in the regional hospital by intramuscular "antirheumatic cocktails". The physical examination revealed a small pulsatile mass over the right buttock associated with overlying bruit. The selective angiography of the internal iliac artery demonstrated a small inferior gluteal artery pseudoaneurysm. The patient was operated by the procedure described. The postoperative recovery and the late result (after 6 months) were good. DISCUSSION: According to our knowledge, only 8 cases of gluteal artery pseudoaneurysms are reported in literature in the last 11 years (including the first three months of this year) [4-8]. The lesions of the gluteal arteries, especially pseudoaneurysms, have no specific symptoms and signs. usually, they appear as haematoma pulsans and neurologic deficiency due to compression. (One of our patients). The gluteal abscess can be a differential diagnostic problem. Duplex ultrasonography, CT and selective angiography can be used in the diagnosis [5]. The standard surgical treatment of gluteal artery pseudoaneurysms consists of the ligature of the internal iliac artery (using transperitoneal or extraperitoneal approach) and pseudoaneurysmal resection and ligation of nutrient vessels by gluteal approach [9]. The second procedure is the temporary clamping of the internal iliac artery and transgluteal ligation of the nutrient vessels [7]. The microcatheter embolization of the nutrient vessels using standard invasive radiologic approaches via femoral artery is the method of choice in the treatment of gluteal artery pseudoaneurysms [10]. A buttock pulsatile mass and neurological deficiency in a patient with history of penetrating gluteal trauma, suggest the existence of gluteal artery pseudoaneurysm and require diagnostic evaluation. PMID- 9863370 TI - [The role of endothelial cells in allergic inflammation reactions]. AB - Inflammatory response in tissue results from a complex network of interactions between inflammatory cells (mast cells, eosinophils, basophils, macrophages) and resident cells belonging to the lung structure (like endothelial cells, fibroblasts, epithelial cells). Among structural cells, endothelial cells play a critical role. The important role of endothelium is also reflected in the fact that it occupies an area exceeding 1000 m2. Thus, endothelium is the largest and the most active paracrine organ in the body, producing potent vasoactive, procoagulant, anticoagulant, and proinflammatory substances. Endothelial cells have four key functions that alter in the process of inflammation: 1 a) Regulation and control of leukocyte traffic through the expression of adhesion molecules (selectins E and P, molecules of immunoglobulin superfamily ICAM-1, ICAM-2, VCAM); 1 b) They are also able to amplify leukocyte activation through the production of proinflammatory cytokines like IL-1, IL-6 and chemokines like IL-8 and RANTES molecules; 2) Regulation of vascular tone by production of PGI-2, EDRF/NO and elements of local renin-angiotensin system; 3) Regulation of local coagulation by controlling the production of t-PA and PAI-1; 4) Regulation of the vascular permeability. In the states of acute inflammation, the endothelial cell takes on a proinflammatory phenotype and as such becomes chemoattractant, facilitating leukocyte adhesion, activation and migration, becomes prothrombotic and demonstrates enhanced vascular permeability. PMID- 9863372 TI - [The modern approach to hysterectomy]. AB - Being quite experienced in the field of gynaecologic surgery and hysterectomies especially, being familiar with recent innovations in laparoscopic surgery and also having some own experience in laparoscopic surgery, the authors discuss the advantages and disadvantages of all surgical methods of hysterectomy. Comparing the techniques, the duration, bearing in mind the the risks, overall costs and all other surgical details, the authors concluded that laparoscopically assisted vaginal hysterectomy is the best choice because it is the least invasive, less risky, no scars are left, the postoperative recovery is quick, there are numerous indications for it, the preparation obtained as a whole can be used for further clinical examination. The only disadvantage is it is too costly and sometimes the operation itself lasts too long, so it should not be applied in some cases. PMID- 9863373 TI - [Arginase activity in plasma and erythrocytes in children with hematologic diseases]. AB - INTRODUCTION: Arginase (EC 3.5.3.1) is one of the essential enzymes in the terminal stages of the urea cycle in the liver which participates in the elimination of ammonia from the human body [1, 7]. Except in liver tissue arginase is also present in many human tissues and in the circulating blood cells, especially in erythrocytes and leukocytes. Arginase splits arginase to urea and ornithine that serve for biosynthesis of amino acid proline, glutamic acid and biosynthesis of polyamines-spermine, spermidine and putrescine. Arginase activity is high during the mitotic cycle, with the function in phase S of the cell cycle. The aim of our study was to assess the arginase activity in the blood of children with some haematologic diseases. METHODS: We examined the arginase activity in blood plasma and erythrocytes of children who suffer from some haematological disorders (27 patients) and in healthy children (control group-15 subjects). The enzyme activity was measured with spectrophotometric method on the basis of the determination of the amount of liberated ornithine from substrate arginine [3]. RESULTS: The obtained results suggest that arginase activity was much higher in the blood of ill children (Table 1 and Figure 1). In the control group of children (total 15) plasma arginase activity was in the range of 0 to 20 U/L x = 0.86 U/L), and enzyme activity in erythrocytes was 1.62-3.98 U/g Hb (x = 2.81 U/g Hb). Erythrocytes enzyme activity and plasma enzyme activity were in ranges of 4.03-5.26 U/L with the mean value of x = 4.56 U/L, and arginase activity in erythrocytes was in ranges of 9.38-14.16 U/g Hb, with mean value x = 11.34 U/g Hb, respectively. Arginase activity in erythrocytes was also significantly higher in children with non-spherocytic haemolytic anaemia (9 children) and was in ranges of 5.33-9.58 U/g Hb (x = 7.29 U/g Hb), with the relatively low values in plasma, 0-4.14 U/L (x = 1.75 U/L). In children with sideropenic anaemia (total number-11) arginase activity in erythrocytes was also very significantly increased with the range between 2.86 and 14.16 U/g Hb (x = 5.54 U/g Hb) while the plasma enzyme activity was relatively low, with the values in range of 0-4.98 U/L (x = 1.41 U/L); in myelodysplastic syndrome (4 pts) arginase activity in plasma was very low (0-0.71 U/L; x = 0.26 U/L) with higher values of arginase activity in erythrocytes (4.22-5.89 U/g Hb; x = 4.94 U/g Hb). DISCUSSION: We have concluded that the enzyme activity was the highest in erythrocyte haemolysates of patients with spherocytosis, non-sherocytic haemolytic anaemia; it was also high, but in a smaller degree, in erythrocytes of children with sideropenic anaemia and myelodysplastic syndrome; arginase activity in plasma of these children was higher in comparison with enzyme activity in plasma and erythrocytes of healthy children. Our results are in agreement with data from literature where it is stated that the younger erythrocytes have the highest arginase activity than the mature erythrocytes [4]. CONCLUSION: The measurement of arginase activity in plasma and erythrocytes is a good diagnostic indicator for the presence of young erythrocytes and reticulocytes in the circulating blood as is the good sign for the detection of haemolytic processes. PMID- 9863374 TI - [Sensitivity to antibiotics in coagulase-negative staphylococci isolated from patients with central venous catheters]. AB - We examined 25 coagulase-negative staphylococci isolated from children, of whom 17 with leukaemia and 8 with terminal renal failure. Strain identification performed by api Staph system revealed the presence of S. epidermidis in 21 children, S. hominis in 3 patients and S. haemolyticus in 1 patient. By diffusion method we examined the activity of penicillin, methicillin, cephalexin, cephtriaxon, lincomycin, erythromycin, vancomycin, co-trimoxasol, gentamicin, amikacin, chloramphenicol, rifampicin and fusidic acid. MICs of seven antibiotics were obtained by agar dilution method. MIC50 and MIC90 were as follows: /ml Methicillin 3.13 mg/ml and 50 mg/ml, lincomycin 100 mg/ml and 100 mg/ml, gentamicin 25 mg/ml and 100 mg/ml, chloramphenicol 6.25 mg/ml and 50 mg/ml, amikacin 1.56 mg/ml and 100 mg/ml, rifampicin 0.09 mg/ml and 12.5 mg/ml, fusidic acid 6.25 mg/ml and 12.5 mg/ml, vancomycin 1.56 mg/ml and 3.13 mg/ml. These data show that the examined strains are highly resistant to numerous antibiotics. Thirty six percent of all strains were resistant to methicillin, 88% to lincomycin, 60% to gentamicin, 52% to chloramphenicol, 24% to amikacin, 52% to rifampicin and 56% to fuscidic acid. All the examined strains were sensitive to vancomycin. PMID- 9863375 TI - [Evaluation of the effect of the health education intervention project "Healthy School"]. AB - Contemporary health-education intervention programs are increasingly used as a tool for improving health of school children [1-4]. Since 1992 a Network of 13 elementary Health Promoting Schools established in Yugoslavia (though not yet admitted to the European Network) has been operational. The Project was approved by the Ministries of Health, Education and Ecology from the very beginning, and financially supported by the Government of Serbia since 1995. The team of up to 40 health professionals, school principals and school project managers worked together for four years to change the working conditions in schools knowledge, attitudes and behaviour of school children and staff in order to improve their health [5]. The goal of this paper is to present results of health education intervention in changing of hygienic conditions in schools, as well as some of the attitudes, behaviour and knowledge of pupils and their parents. METHOD: The study took place before and after the intervention--at two points in time--during 1993 and 1996. The tri-angular approach including (1) pupils, (2) schools (teachers, school environment), and (3) parents, was used. A random pretest and post-test study design with control group (12 experimental and two control schools) has been implemented. The multiphase cluster sample was employed in order to represent all of the country typical regions. Six types of especially designed questionnaires were used to provide comparable variables in the sample of pupils, their parents and teachers. Exception were 1st-graders and 4th-graders for whom information were gathered by means of a "draw-and-write" investigation technique [6]. The response rate was 88.70% before and 98.28% after intervention. Data were analyzed by descriptive and inferential statistics using SPSS/PC software. RESULTS: Schools are somewhat less overcrowded, much cleaner and better maintained after the four-year intervention. Toilets are in a better condition, but there is still much more to be done. The 1st- and 4th-graders, surveyed by the draw-and-write method, mentioned numerous ways of keeping and improving health, which were summarized as fifteen health-improvement measures (Graph 1). The most frequently mentioned measures were nutrition, physical activity, general hygiene, oral and dental hygiene, sleeping and fresh air. Each of these measures was mentioned by over 20% of the surveyed pupils. Eleven of 15 variables showed significant increase in frequencies (at the level of at least p < 0.01) after the intervention. As an indicator of a nutrition behaviour, the regularity of main meals is analyzed (Graph 2). The majority of children eat regularly and the difference before and after intervention is significant only for the school meal (c2 = 30.715, p < 0.001). Although over 70% of children learn that general hygiene is good for health in junior graders, only about little more than 30% of senior graders have a bath or shower every day, while others only once or twice a week. The differences are significant before and after the intervention (c2 = 10.648, p < 0.05) only for everyday practice. More than 90% of senior-grade pupils brush their teeth at least once or several times a day. Over 60% of children in our survey go in for sport, whereas about 20% never do so. It seems that the intervention contributed symbolically to this practice important for health, though before-after difference is significant (c2 = 6.673, p < 0.05). However, in the control group schools children have much less physical activity in 1996, and this difference is significant (c2 = 14.070, p < 0.010). The psycho emotional status of Yugoslav young people is strongly influenced by the situation in the country the war, the economic disaster and the impact of international sanctions. Consequently, more than one-fourth of the children complained of frequent exhaustion and concentration problems, which their parents also noticed. (ABSTRACT TRUNCATED) PMID- 9863376 TI - [Therapeutic approach in blast injuries of the ear]. AB - Otic blast injury is caused by arrhythmic air blast wave. The perforation of the tympanic membrane is the commonet finding associated with lacerations of mucosa in the middle ear. Makki [6] published 34 cases of myringoplasty after war blast injuries. However, healing of such perforations is common; Kerr [7] noted a healing rate of 83% after the blast injury. The aim of the study was to evaluate different therapic procedures of otic blast injuries. There were 74 patients with bilateral otic blast injuries, who underwent otomycroscopic examination. The following parameters were noted: Integrity of tympanic membrane and size of perforation, presence of haemorrhagic exudate in the middle ear and median value of conductive deafness (Table 2). Main symptoms, presented in Table 1 were: pain, deafness and otohaematorrhoea. The first group of 19 patients received antibiotics by parenteral way during 7 days according to the bacterial finding in ear exudate. Healing rate was dependent on the size of tympanic membrane perforation, and rated from 71% in perforation of one third of tympanic membrane to 25% in total perforation of tympanic membrane. Data are presented in Table 3. The second group of 24 patients received the same therapy as the first treatment, plus otomycroscopic removal of haemorrhagic exudate, lacerated middle ear mucosa, and repair of tympanic membrane lacerations. Healing rate was significantly better than the one obtained in the first group (Table 4). Persistent tympanic perforation, as indicator of failed therapy, was present in 5 (35%) of all examined ears with two thirds of tympanic membrane perforation in the first group, while in the second group the rate was 3 (12%). The third group received the same therapy as the second treatment, including administration of amicacyn into the external auditory canal. The results were statistically compared by chi 2 test, and we found that the second therapy protocol was significantly better. There was no significant difference between the second and the third therapy protocols. We found blast ruptures of tympanic membrane and auditory ossicles chain discontinuity in 88% of examined ears. In our material the high incidence of total tympanic membrane ruptures and subtotal ruptures (48%) is quite different in comparison to other authors [5-7]. We consider it as the effect of high power blast wave. Loss of conductive hearing was present in 91% of blast injured patients, while 7.4% of patients had mixed, predominantly senzoneural deafness. Consequently, in addition to mechanic blast injuries acoustic trauma could profoundly damage the inner ear. Spontaneous healing of tympanic membrane occurred in 71% of injured persons and this was a better result than the results obtained by other authors [3, 4, 6]. The spontaneous healing of tympanic membrane failed if infection of the middle ear occurred or blast caused the total tympanic membrane perforation. In the second and third therapy protocols significantly higher healing of tympanic membrane ruptures was evident; it rated from 88% to 91%. Better results could be explained by the effect of optimal healing conditions, based on removal of exudate from the middle ear and necrotic parts of tympanic membrane, completed by fitting of lacerated parts of tympanic membrane and antibiotic prophylaxis. Healing of tympanic membrane without scars and adhesions was more frequent than in patients treated only with antibiotics. Incidence of undesired outcome of persistent perforation of tympanic membrane was reduced. To prevent posttraumatic complications in the middle ear, we recommend early cleaning of margins, reposition of lacerated fragments of tympanic membrane, and removal of haemorrhagic exudate. Myringoplasty should be performed if spontaneous healing of tympanic membrane did not occur after 6 months. CONCLUSION: Otic blast injury was frequently found in war induced trauma. (ABSTRACT TRUNCATED) PMID- 9863377 TI - [Chronic rupture of abdominal aortic aneurysms]. AB - INTRODUCTION: Rupture of abdominal aortic aneurysms (RAAA) can take place in one of the 4 following ways: 1. "Open" rupture in the free peritoneal cavity; 2. "Closed" rupture with formation of retroperitoneal haematoma; 3. Rupture into surrounding cavity structures, such as veins and bowels; 4. In rare cases rupture is effectively "sealed of" by the surrounding tissue reaction, and retroperitoneal haematoma is "chronically" contained [1]. The terms "sealed" [2], "spontaneously healed" [3], "leakig" [4] RAAA, were also used in the previous papers connected to this situation. The "sealed" rupture was first described by Szilagyi and associates in 1961 [2]. In their case the rupture was small and haemorrhage was effectively encircled by the tissue surrounding the aortic wall. The slow rate of blood loss contributed to the patient's haemodinamically stable condition. Christenson et al. reported a case of "spontaneously healed" RAAA [3]. Rosenthal and associates described 2 patients who had aortic aneuryms that ruptured several months before repair and contributed to the term "leaking AAA" [4], while Jones et al. introduced the term "chronic contained rupture" [1]. The aim of this paper is the presentation of 5 such patients. CASE REPORT: Between December 1, 1988 and May 30, 1997 411 patients with abdominal aortic aneurysms (AAA) have been operated at our institute. Of this number 137 (33%) had RAAA, while 5 patients (12%) had a contained RAAA (CRAAA). CRAAA were found in 3 male and two female patients, average age 62 years. All of them had a previously proved AAA and initial symptoms lasted for days or months before the admission. In all patients haematocrit, pulse rate and arterial tension during the admission, were normal. All typical signs of RAAA were absent in these patients. Patient 1. A 56-year-old man, smoker, with previous history of arterial hypertension had an isolated episode of abdominal pain and collapse 30 days before the admission. Physical examination revealed a pulsatile abdominal mass. Doppler ultrasonography identified an infrarenal AAA, with right lobular extraaneurysmal mass which displaced the inferior vena cava (ICV). Angiographically (Figure 1a) an unusual saccular intrarenal AAA was detected, while simultaneous cavography (Figure 1b) confirmed the-dislocated inferior vena cava to the right. The intraoperative finding showed infrarenal CRAAA with organized retroperitoneal haematoma between AAA, ICV and duodenum. After aortic cross clamping and aneurysmal opening, the rupture at the right posterior aneurysmal wall was discovered. The partial aneurysmactomy and aortobilliar bypass procedure with bifurcated knitted Dacron graft (16 x 8 mm), were performed. The patient recovered very well. After a 4-year follow-up period the graft is still patent. Patient 2. A 72-year-old woman with low back pain, fever and disuric problems was urgently admitted to the Institute of Urology and Nephrology. The standard urological examination (X-ray, intravenous pyelography, retrograde urography, kidney Duplex ultrasonography) excluded urological diseases. However, intrarenal AAA an a giant aneurysm of the right common iliac artery, were found. The proximal dilatation of the right excretory urinary system was also found by retrograde urography. The patient was transported to our Institute 20 days after the initial symptoms. Translumbar aortography (Figure 3) showed the right common iliac artery aneurysm and gave the false negative picture of normal abdominal aorta because of parietal thrombosis of AAA. The intraoperative finding showed chronic rupture of the posterior wall of the right common artery aneurysm. The retroperitoneal haematoma compressed the right ureter. Both aneurysm have been resected and replaced by bifurcated Dacron graft (16 x 8 mm). The patient recovered successfully. After a 2-year period of follow up the graft is still patent. Patient 3. (ABSTRACT TRUNCATED) PMID- 9863378 TI - [Radical hysterectomy with pelvic lymphadenectomy in patients with carcinoma of the uterine cervix--3 years' experience]. AB - INTRODUCTION: Almost all patients with invasive cervical carcinoma can be treated with either primary irradiation therapy or primary surgery. Some patients are appropriately treated with the combination of irradiation and surgery. Chemotherapy is not effective as primary treatment of invasive cervical cancer but may be used as additional therapy and when the disease is recurrent or persistent. There are some important advantages of primary extensive surgery over irradiation. The findings at operation and that from the careful pathologic examination of surgical specimens can be very helpful in selection of patients for supplementary postoperation irradiation therapy or chemotherapy, or both [1 6]. AIM OF THE STUDY: The aim of the study was to compare pretreatment clinical evaluations with surgical and postsurgical pathohistological findings. METHODS: Extensive hysterectomy and bilateral pelvic lymphadenectomy were used in the treatment of 251 patients with early invasive cervical cancer. The patients were treated at the Department of Obstetrics and Gynaecology of the Clinical Centre of Serbia in Belgrade, between 1993 and 1995. Cervical cancer was detected by clinical examination, colposcopic and cytologic (Pap smear) findings, colposcopically directed biopsy or conisation and pathological findings, sonography, chest radiography, blood and urine analyses. In some cases we had to make other examinations (cystography, cystoscopy, intravenous pyelography, sygmoidoscopy, rectoscopy, CT scanning and magnetic resonance). The surgical treatment of invasive carcinoma of the cervix was limited to those patients in whom the disease was confined to the cervix or vaginal fornix (stage Ia, stage Ib or stage IIa), and who were in high surgical risk. RESULTS: Over a three year period (1993-1995) there were 251 patients with invasive cervical cancer, treated by primary surgery (radical hysterectomy and bilateral pelvic lymphadenectomy sec. Werthein-Meigs), average age 42 years. Most of the patients demonstrated invasive cervical cancer, clinically classified in Ib st. (81.67%). Some characteristics of pathologic findings, such as parametric width, number of removed lymph nodes, percentage of lymph nodes metastases and correlation with clinical stage of invasive cervical cancer, histologic grade of cervical cancer with lymph node metastasis, pathologic findings after surgical treatment, correlation between clinical and surgical staging, were already presented in tables. DISCUSSION: In the last decades the incidence of invasive cervical cancer and death rate have been decreased. Progress in reducing mortality is primarily attributed to the introduction of cervical cancer screening as part of regular gynaecologic examinations. Regular testing with Papanicolaou (Pap) smear and colposcopy have an important role in this problem [1]. Extensive hysterectomy and bilateral pelvic lymphadenectomy were used in the treatment of 251 patients with early invasive cervical cancer. We found that the clinical diagnosis of disease extent was correct in 67.7% of patients who underwent extensive surgery for early invasive cervical cancer. Sensitivity of clinical findings was 75.8% and positive predictive value was 86.2%. Lymph node metastasis was detected in 17% patients. Brodman at al. [14] found that clinical examinations, including CT scanning and magnetic resonance, were correct in only 62.5% of cases. It is very difficult to detect parametric involvement and lymph node metastasis by clinical examinations. Irradiation therapy was used in the postoperative period as additional treatment of extensive hysterectomy and bilateral pelvic lymphadenectomy in 89.7% of patients. CONCLUSION: The findings at operation and that from the careful pathologic examination of surgical specimens are absolutely irreplaceable and important in grading invasive cervical cancer and selection of patients for supplementary postoperate irradiation therapy. PMID- 9863379 TI - [Primary tumors of the fallopian tubes: retrospective analysis of results in 7 patients]. AB - Primary malignant tumours of the Fallopian tube are among the rarest of gynaecological malignancies. Seven patients with primary malignant tumour of the Fallopian tube, treated between 1991 and 1996, were studied. The average age was 60 years, with nulliparity rate of 29 percent and a mean parity of 2.2. The most common symptoms were atypical vaginal bleeding (29 percent), abdominal pain and distention (29 percent), whereas 43 percent of patients had no symptoms. No patient had a correct preoperative diagnosis. Primary surgical treatment was in all patients hysterectomy and bilateral salpingo-oophorotomy with or without omentectomy. Staging was done according to the FIGO classification for Fallopian tube carcinomas. In Stage I 29 percent of patients were classified, 43 percent in Stage II and 29 percent in Stage III. Six patients (86 percent) had adenocarcinoma (1 in G1, 2 in G2, 3 in G3) and 1 had MMMT (malignant mixed tumour heterology type). All patients received additional postoperative therapy including: chemotherapy (58 percent), radiotherapy (14 percent), hormonotherapy (14 percent) or combination of chemotherapy and hormone therapy (14 percent). Our results are comparable to those of other authors. The time is too short to predict a 5-year survival, but this will be reported in our next paper. PMID- 9863380 TI - [Adhesion molecules in kidney diseases (part I)]. AB - Adhesive molecules are (glyco)proteins of the cellular membranes. All of them have their extramembranous, transmembranous and intracytoplasmatic parts. As receptor molecules, their extracellular parts bind the specific ligand. The ligand can be found on the surface of the other cell or in the extracellular matrix (basal membranes). The following families of adhesion molecules are: cadherins, selectins, integrins and members of immunoglobuline supergene family. Different members of the same family could have different times (in ontogenesis, in adult form) and space distribution (in different tissues, different tissue structures). The contact between the cells and basal membranes with these molecules is important for cell division, maintaining the tissue architecture, polarization and function of cells, migration of cells, endo- and exo-cytosis as well as for maintaining the structure and function of basal membranes. As above stated all this is important in the occurrence morphogenesis, haemostasis, inflammation, malignant cell transformation and metastasis. This knowledge is important for the better understanding of renal diseases. PMID- 9863381 TI - [Toxoplasmosis and immunosuppression]. AB - Toxoplasmosis is a widely distributed zooanthroponosis, caused by the ubiquitous obligatory intracellular protozoan parasite Toxoplasma gondii. Once infected, the host acquires lifelong immunity induced by the persistence of the parasite in an encysted form. While T. gondii infection in pregnancy has long been known as a significant cause of perinatal morbidity and mortality (congenital toxoplasmosis), its significance as an opportunistic agent has been increasingly recognized during the last decade, particularly with the outbreak of AIDS. Reactivation of a previously latent infection results in a wide clinical spectrum, predominantly within the central nervous system. The paper reviews recent data on the significance of toxoplasmosis as an opportunistic infection in immunosuppressed individuals, such as patients with malignant and systemic diseases treated with immunosuppressive drugs, organ transplant recipients, and, first and foremost, patients with AIDS. A high prevalence of latent toxoplasmosis in Yugoslavia indicates a high local exposure to infection reactivation. While a definitive diagnosis of toxoplasmosis is difficult in the immunosuppressed, its treatability as opposed to a fatal outcome, if untreated, demands that physicians caring for the above categories of patients keep in mind toxoplasmosis and its possible clinical presentations and include them in the differential diagnosis of these conditions. PMID- 9863382 TI - [Epithelial cells in bronchial asthma]. AB - The paper deals with the current data on the morphologic and functional damages to the epithelium in asthma. Structure of the normal airway epithelium is briefly described. Damage to the epithelium disrupts the barrier to the external environment, and may result in changes in mucous secretion, and mucociliary clearance; induce bronchoconstriction; promote chemotaxis of inflammatory cells. Epithelial cells can present antigens to T-lymphocytes. Epithelium is both target and effector in asthma. Hypothesis that eosinophils contribute to the damage of the epithelium gained increasing recognition. PMID- 9863383 TI - [Neurologic manifestations in trichinosis]. AB - Neurologic manifestations are present in about 10-20 percent of patients with trichinosis. They could be a serious diagnostic problem in the absence of corresponding epidemiological data and typical symptoms and signs of the disease. In untreated patients the mortality rate is about 50%. Several pathogenic mechanisms are responsible for the neurological complications in trichinosis: obstruction of brain blood vessels by larvae, cysts or granulomas, toxic vasculitis with secondary thrombosis and haemorrhages, granulomatous inflammation of the brain parenchyma and allergic reaction. Neurotrichinosis is manifested with clinical symptoms and signs of meningitis, encephalitis, polyradiculoneuritis, poliomyelitis, myastenia gravis, paresis and paralysis, with the clinical picture of systemic disease of the connective tissue involving the nervous system and, extremely rare, as a sinus thrombosis. Thus, the broad spectrum of neurological lesions in trichinosis is, probably, the results of the fact that Trichinella spiralis larvae, during haematogenic dissemination has no special affinity for particular parts of the nervous system. We present five patients with encephalitis and focal cerebral lesions in trichinosis. In one patient the neurologic manifestations were the only sign of the disease. We believe that all pathogenic mechanisms mentioned above, were involved in the onset of neurological manifestations in our patients. The diagnosis of the disease was based on the clinical picture, epidemiological data, microscopic identification of larvae in the muscular tissue, the presence of antibodies against Trichinella spiralis in cerebrospinal fluid (with preserved blood brain barrier) and in serum confirmed by IIF method, computerised tomography and magnetic resonance imaging of the brain, eosinophilia in the peripheral blood picture. One patient died, and in the remaining patients the course of the disease was favourable; they were discharged from the hospital with minimal neurologic sequelae. PMID- 9863384 TI - [An ultrasonic scalpel for laparoscopic gynecologic surgery]. AB - In order to obtain good results in laparoscopic surgery the surgeon should be well trained and experienced and the equipment perfect. The recent innovation in laparoscopic surgery is the ultrasonically activated scalpel. Scissors make the working part of scalpel with one hand vibrating 55,000 Hz in a second. The effect reflects in braking hydrogen links and denaturation of proteins in instant haemostasis and coagulation in blood vessels, and there is no need of tying them up. Practically, there is no damaging of the surrounding tissues. The technique is easy to use. When performing laparoscopic vaginal hysterectomy and adnexectomy the ligamentum infundibulopelvicum and ligamentum rotundum are cut, and there the blood vessels are not thicker than 3 millimeters. As these ligaments are easy to reach with ultrasonically activated scalpel, we can say that this instrument is the ideal option for laparoscopic vaginal hysterectomy. PMID- 9863385 TI - [Anniversaries of the Serbian Medical Society. 60 years since its founding. The dream is fulfilled--the home of the Serbian Medical Society is opened]. AB - The Serbian Medical Society was founded in Belgrade in 1872, 126 years ago. At that time, Serbia was liberated from the Ottoman domination, and was one of some thirty existing independent states gaining international recognition in 1878. In 1932 an old dream has been fulfilled--on the occasion of the 60th anniversary of the existence and activity of the Serbian Medical Society--the home of Serbian Medical Society was opened. A 30-year-long period in which the building of the Home was one of the main preoccupations and a "guiding light" of Serbian doctors thus ended. Money from charitable funds was used, which caused certain benefactors to be praised as noted personalities. Medical practice in those days was in the state of choice. There were no means for adequate treatment of certain ailments, and therapy without realistic scientific base was given, often covered by fictitious reasons. This was especially true for tuberculosis and cancer. Under the pretext of the introduction of "new therapeutical approaches" into medical practice and treatment, diverse pharmaceutical formulas were introduced without knowledge of their real effect--injections of milk, drug containing animal embrional cells and special attention was paid to transplantation of the sexual glands. The injection of "camphor oil" (5 cc) was thus "recommended, harmless but useless". The treatment of tuberculosis, which domineered the pathology of population, was very chaotic. The greatest number of drugs for "successful cure" were to be found here. Most commotion was caused by the so called Friedman's cure for tuberculosis which was rejected only after vigorous debates. Our drug "Joannin" on the basis of "the old tuberculine" was also represented in this confusion (and Koch himself was forced to recant it). This medicament was also hailed as "successful cure". The origin of serious scientific efforts, however, are to be found around newly formed journal "Medicinski pregled", which attracted new and progressive contributors. At this time, the newly formed Ministry of Public Health started a campaign for the introduction of a modern organization of the health care and the inclusion of all population in it. However, in a country devastated by war and stricken by poverity, annomalies in the functioning of the health care existed, as financial preconditions did not exist and healt insurance was still in its infancy. This made the status of doctors difficult, causing long debates in the Serbian Medical Society and fierce criticism of the Ministry of Public Health. The responsible persons were accused of introducing and promoting communist ideas and revolution under the duise of health care for the poipulation. PMID- 9863386 TI - [Renal tubular function in children with hypercalciuria]. AB - INTRODUCTION: Renal stone disease is commonly due to hypercalciuria [1, 2], which may be assessed either from a 24-hour urinary collection or from the fasting first morning urine. Hypercalciuria during childhood has been defined by a 24 hour calcium excretion greater than 3.5 mg/kg per day and/or calcium to creatinine ratio greater than 0.20 [3]. The alteration in the calcium transporting systems plays a pathogenetic role in promoting hypercalciuria [4, 5]. Since calcium reabsorption along the nephron is intimately related to that of other electrolytes and substances, it can be hypothesized that patients with hypercalciuria may have other renal tubular defects. The aim of the study was to investigate proximal tubular function (tubular reabsorption of sodium, potassium, phosphate and glucose) and distal tubular function (urinary concentrating capacity and acidifying capacity) in children with hypercalciuria. PATIENTS AND METHODS: Two groups of children were studied: hypercalciuric group included 23 children with hypercalciuria (10 males, aged 11.9 +/- 4.1 years), of whom 6 with nephrolithiasis, and control group included 42 healthy children (20 males, aged 11.2 +/- 3.8 years). All subjects had normal serum values for calcium, sodium, potassium, phosphate and glucose, as well as normal renal function. The urinary excretion of calcium, sodium, potassium, phosphate, glucose and creatinine was measured in a 24-hour urine specimen by standard laboratory methods. Urine osmolality and urinary specific gravity were measured following 12-hour water deprivation test. A short ammonium chloride loading test was performed in 3 patients with urinary pH above 5.5. The fractional excretion of sodium, tubular phosphate reabsorption and renal threshold phosphate concentration were calculated according to standard formula. Statistical analysis was performed using the t-test and analysis of variance (ANOVA). Kruskal-Wallis method was used to compare urinary phosphate excretion between two groups. RESULTS: Table 1 summarizes urinary excretion of electrolytes in children with hypercalciuria compared with healthy controls. We found that urinary sodium excretion was significantly increased in patients with hypercalciuria when compared with controls (p < 0.05). Urinary phosphate excretion was significantly higher in patients with hypercalciuria in comparison to controls, and this was accompanied by a significant lowering of the tubular phosphate reabsorptive threshold (p < 0.05). Urinary potassium excretion tended to be lower, although not significantly, in the hypercalciuric children than in normal subjects. Table 2 shows the mean values +/- standard deviation of urinary specific gravity, urinary osmolality and urinary pH. Urinary specific gravity mean value was significantly lower in patients with hypercalciuria in comparison to controls (p < 0.05). Urinary pH was found below 5.5 in all patients. Glycosuria was detected in 3 patients (13.3%). As shown in Graph. 1, a significant correlation between the urinary excretion of calcium and sodium was demonstrated in both groups of children (r = 0.29; p < 0.01). DISCUSSION: The present study shows that children with hypercalciuria have significantly higher urinary sodium and urinary phosphate excretion in comparison to controls, while urinary potassium excretion is normal in both groups of children. According to some recent reports [6-9], these findings may indicated defects of the renal tubular transport of sodium and phosphate which may be interpreted as a cause or a consequence of the alteration of the calcium transporting system. Defects in both proximal and distal renal tubular functions have been demonstrated in patients with nephrolithiasis, particularly those with hypercalciuria. Proximal renal tubular defects include defects in sodium, fluid, phosphate and glucose reabsorption, which were evident also in our patients. (ABSTRACT TRUNCATED) PMID- 9863387 TI - [Cystic degeneration of the tunica adventitia of the popliteal artery]. AB - INTRODUCTION: Adventitial cystic disease of the popliteal artery (PA) is an uncommon and unique entity characterized by a mucinous cyst located in the arterial adventitia. As the cyst enlarges, it provokes vascular compression with stenosis or occlusion, the first only during the knee flexion, and then in all leg position. Atkins and Key (1946) were the first who described this disease in the external iliac artery [1]. Eirup and Hiertonn (1956) described the disease in the PA, which is the place of its most common localization. The aim of the paper is the presentation of our 10 cases of PA adventitial cystic disease. PATIENTS AND METHODS: Ten patients with PA adventitial cyst were treated at the Institute of Cardiovascular Diseases of the Serbian Clinical Centre in Belgrade, over the period between 1978 and 1997. There were 9 males and one female patient, average age 42.7 years (31-62). Two patients were smokers, while all other atherosclerotic risk factors, including heart disease, were absent. The diagnosis was established using Doppler ultrasonography and angiography. The postoperative histological examination revealed PA adventitial cyst in all patients (Figure 1). In Table 1 are presented our patients. The patients 3 and 4 were admitted for acute ischaemia of the leg. In patient 3 Doppler indexes were 0.0, and transfemoral arteriography revealed segmental occlusion of the PA. All other arteries were unchanged. These findings suggested an unusual disease of the PA. During the operation the posterior approach to the PA was used, and intraoperatively the adventitial cyst was found. In patient 4 the tibioperoneal trunk, posterior tibial artery and PA were occluded. Therefore, the medial approach to the PA was used. After thrombectomy of the crural vessels, the popliteo-popliteal bypass procedure was performed. The PA resection by this approach was not possible. The ringed 6 mm PTFE graft was used for reconstruction because of inadequate saphenous vein. The patients 1, 2, 5-10 were admitted with disabling claudication discomforts. In patients 1, 2, 5, 6, 8, 9 popliteal and pedal pulses were absent, and Doppler indexes decreased. In patients 7 and 10 pedal pulses were palpable and decreased during the normal knee position, while absent during the knee flexion. During some maneuvers Doppler indexes significantly decreased. Transfemoral arteriography in patients 1, 2, 5, 6, 8, 9 showed segmental stenosis or occlusion of the PA, and for this reason the posterior approach to the PA was used. The PA adventitial cyst was found in all cases (Figure 2). In patient 7 angiography revealed a "hourglass" deformity of the PA, while in patient 10 "scimitar" sign was found. Both angiographic findings are characteristic of PA adventitial cyst. The posterior approach was carried out in all patients. In patient 2 only cyst aspiration has been performed, while in patients 7, 8, 9 aspiration and resection of the changed PA adventitia (Figure 3a, figure 3b). In patients 1, 3, 5, 6, 10 an occluded arterial segment was resected. The restoration of the flow observed after the end-to-end anastomosis in patient 1, and after interposition of the saphenous graft in other patients. After the operation, the contralateral leg was examined using Doppler ultrasonography in all patients. The Doppler indexes were significantly decreased in patients 1 and 5 during the knee flexion, but the patients refused the angiographic examination. The control examination consisted of physical examination, Doppler ultrasonography and sometimes angiography; it was carried out after 1, 3, 6 and 12 months, and then every year after the operation. RESULTS: There was no mortality among our patients in the early post-operative period. In patients in whom cyst aspiration was performed, claudication discomfort was decreased, and Doppler indexes were significantly increased. In patients with arterial resection and reconstruction (1, 3, 4, 5, 6, 10) the effect of the operation was simi PMID- 9863388 TI - [Aplastic anemia--clinical characteristics and survival analysis]. AB - Aplastic anaemia is a relatively rare disease, characterized by the loss of haemopoietic "stem" cells in the bone marrow, exchanged with fat cells and pancytopenia. The treatment of this disease consists of supportive therapy with blood products and finally phase, where there are two possibilities: immunosuppression and bone marrow transplantation. Immunosuppression means the use of antilymphocyte or antithymocyte globulins, anabolic steroids, Cyclosporine A, corticosteroids, and recently haemopoietic growth factors [1]. The advantage of bone marrow transplantation compared to other kinds of treatment has been established in multicentric trials [2]. Goals of study: 1. To show and analyze clinical characteristics in patients with aplastic anaemia; 2. To study survival according to gender, age and kinds of treatment. METHODS: Thirty three patients with aplastic anaemia are included in the study. They were treated at the Institute of Haematology, Clinical Centre of Serbia, from 1988 to 1995. For description of data we used parameters for the arithmetic mean and standard deviation; for nonparametric tests-median. Student's T-test was used for comparison of differences among the mean values from nonparametric analytic models. From nonparametric models, Fisher's test was used. For data analysis of survival Kaplan-Mayer's model was used. RESULTS: Thirty three patients with aplastic anaemia were examined. There were 17 (52%) men and 16 (48%) women. The youngest patient was 15 and the oldest 73 years. The average age of patients was 40.18 years. In the group up to 30 years there were 16 (48%) patients. In the group from 31 to 45 years there were 4 (12%) patients. In the group from 46 to 60 years there were 4 (12%) patients, and in the group from 61 to 75 years 9 (28%); clinical signs (fatigue, exhaustion) and anaemic signs (pallor) were present in all patients (33). In 12 (36%) patients high body temperature was established and in 21 (64%) patients there was no temperature. Signs of haemorrhagic syndrome were established in 26 (79%) patients and 7 (21%) patients had no such signs. No significant statistical difference in survival (Lee-Desu = 0.071; DF = 1; p = 0.789) related to sex, was observed. According to the duration of the disease by year, and over this period, the difference was statically very significant (Lee Desu = 20.735; DF = 1; p = 0.000). Therefore, in patients with a longer duration of the disease than one year the chance for a longer survival is better. We compared the group of patients treated with antilymphocytic globulin with the group treated with other methods (androgenic hormones and Prednisolone, Cyclosporine A and bone marrow transplantation). Between these two groups the difference was statistically significant (Lee-Desu = 2.742; DF = 1; p = 0.018). Therefore, survival of patients of the second group was longer thanks to a successful bone marrow transplantation. The group of patients treated by bone marrow transplantation was compared with the group of patients treated with other three methods; the difference was statistically significant (Lee-Desu = 7.346; DF = 1; p = 0.006). The best method of treatment is bone marrow transplantation in comparison to the other three methods. Survival was also analyzed in all four studied age groups. No statistically significant difference was found. CONCLUSIONS: Aplastic anaemia is a serious, relatively rare illness, appearing with almost equal frequency in men and women. Two peaks were noticed: in the younger age group up to 30 years (48%) and over 60 years (27%). According to survival, there was a group with short survival, up to 12 months, and a group with longer survival, over 12 months. Gender and age had no influence on duration of survival. Only the duration of the disease had a positive effect on survival. In comparison to other methods, the most efficacious treatment was allogenic bone marrow transplantation. PMID- 9863389 TI - [Secondary cataracts in extreme myopia]. AB - INTRODUCTION: After cataract surgery there is a higher incidence of posterior capsular opacification (secondary cataract) in myopia than in normal population, and it varies from 40 to 60% [1, 2]. METHODS: Fifty myopic eyes operated for cataract at Prof. Dr. Ivan Stankovitsh Eye Department from 1988 to 1995 were examined for capsular opacification (secondary cataract). Extracapsular cataract extraction was performed in all patients and in 32 patients the posterior intraocular lens was placed. Capsular opacification was classified by type and treated with Nd:YAG laser or discussion at least 9 months to 3 years and 3 months from the operation. RESULTS: There were 13 secondary cataracts (26%) in our series of patients, of whom 22% with intraocular lenses, and 33% were aphakic patients. Visual acuity was 4/60 to 0.5. In 11 patients YAG-laser capsulotomy was performed and in one patient capsular discussion, and in one subject there was no treatment. Consequent visual acuity was 0.3 to 1.0. No correlation was found between the age, type of intraocular lens and capsular opacification. DISCUSSION: Capsular opacification is more often seen in myopic eyes and its treatment is associated with a higher rate of complications (cystoid macular oedema, rupture of anterior hyaloid, retinal detachment...) [3, 4]. It seems that posterior lens implants reduce the chance for development of posterior capsular opacification. The safest treatment is YAG-laser capsulotomy. PMID- 9863390 TI - [Clinical importance of tympanometry in the diagnosis of chronic secretory otitis]. AB - Secretory otitis media is defined as a fluid in the middle ear without signs or symptoms of infection. As the aetiology and pathogenesis of the disease are unknown, and as it affects children aged from 3 to 12 years, treatment procedures proposed for management of secretory otitis media, are not uniform. Some authors [1, 4, 6] consider that functional or mechanical obstructions of the Eustachian tube could provoke secretory otitis. The purpose of the treatment is to remove exudate from the middle ear and appropriately ventilate it for a longer period. That could instantly normalize the hearing and exclude the appearance of late complications of secretory otitis. Although the disease could heal spontaneously, the treatment should be performed immediately for preventing sequelae of secretory otitis. The aim of the study was to evaluate possible aetiologic factors of secretory otitis in our population, and to evaluate results of lympanometry in children with exudate in the middle ear. There were 65 children, aged from 3 to 12 years (Table 1), who complained of deafness and were examined at the ORL Department in Banja Luka. The clinical examination revealed the integrity and color of tympanic membrane, scars, adhesions and atrophic areas. Audiometry and tympanometry had been performed in addition. Patients who proved to have exudate in the middle ear received nasal decongestants and mucolitics during three months, and were evaluated every three weeks by audiometry and tympanometry. Pathologic findings in the nose and epipharynx were the most common findings: enlarged adenoids in 38 (58%) patients, hypetrophic rhinitis in 15 (23%) and allergic rhinitis in 5 (8%) patients. Frequent relapses of middle ear infection in the first three years of life were found in 26 (40%) patients and early first attacks in the first year of life in 15 (23%) patients (Table 2). Premature onset (15%) and allergy (21%) had also been frequently found. Results of tympanometry and audiometry are shown in Table 3. Exudate in the middle ear and type B tympanogram were found in 86 ears, while in other patients dysfunction of the Eustachian tube and type C1 and C2 tympanograms were found. After 6 weeks the exudate disappeared in 16 ears and tympanogram converted in type A and type C2, while the initially found C1 tympanogram was transformed in type A in 5 of 13 ears. After 12 weeks the tympanogram type B was found in 46 ears, while in 40 ears (47%) the tympanogram was changed in type A and type C2. After 6 and 12 weeks of therapy tympanometric types were statistically examined by chi 2 test. We have found a significant difference in tympanometric types and prevalence of type A and C1 tympanograms. Paracentesis and insertion of ventilating tubes were done in 46 ears with the remaining exudate. We have found mucous exudate in 35 (76%) ears associated with retraction and scars of tympanic membrane (Table 4), what indicated that the longer duration of mucous exudate caused degenerative changes in the middle ear. Serous exudate, found in 9 ears (24%), did not affect the color and integrity of the tympanic membrane. Sensitivity of tympanometry in detection of exudate in the middle ear was 96%. Secretory otitis media is a frequent disease in childhood, that could cause functional and morphological sequelae in the middle ear. As for now, there is no unique concept of diagnosis and treatment of the disease, and it is still a current problem. We suggest a three-month evaluation of tympanometric and audiometric patterns, repeated every three weeks, in children suspected of having exudate in the middle ear. There is a large trend of spontaneous disappearance of exudate in the middle ear and changing of tympanogram type. Such children should be evaluated over the period of one year, and if there is no relapse additional treatment should not be carried out. If exudate in the middle ear persists for three months and type of the tympanogram is unchanged, myringotomy and insert PMID- 9863391 TI - [Phonosurgical therapy of nodular lesions of the vocal cords]. AB - INTRODUCTION: Nodular lesions of the vocal fold include contact oedema, contact thickenings and vocal nodules [1, 2], which all appear at the predilection sites of the vocal folds (junction of the anterior and middle thirds, slightly below the free edge). All nodular lesions develop as a consequence of acute or prolonged voice overuse or misuse. Contact oedema is characterized by an intact epithelium and subepithelial oedema (Fig. 1), while contact thickenings present somewhat thickened epithelium and subepithelial oedema. Vocal nodules are characterized by marked intraepithelial thickening, in conjunction with a certain degree of hyperkeratosis. MATERIAL AND METHODS: A series of 293 nodular lesions of the vocal fold was operated by the use of direct microlaryngoscopy (DML), indirect microstroboscopy (IMS) and indirect videostroboscopy (IVS). Based on the functional surgical results, indications were established for each of the surgical technique used. Corticoids were used postoperatively, while the voice therapy started 3 weeks after surgery. RESULTS: During a 10-year-period 1550 surgical procedures were carried out for benign lesions of the vocal folds, including 293 operations for nodular lesions (18.9%). Surgery for vocal nodules was conducted in 268/293 overall operations for nodular lesions (91.5%). Indirect surgery was used in 221 patients (83.9%). It was established for indirect surgery, especially IVS, that it is a method of choice for the surgical treatment of nodular lesions. In some cases it is also suitable for unfavourable local anatomy and contraindications for general anaesthesia which is required for direct operations (DML). Indications for IVS and IMS were identical. DML was used according to a specific wish of the patient, or in cases when topical anaesthesia was not sufficient to carry out the procedure (about 3% of overall number of patients). Recovery of the vibration pattern was faster and more complete after indirect operations, especially after IVS (Table 1). DISCUSSION: Indirect videostroboscopic procedure is the best surgical choice for nodular lesions of the vocal fold. It is characterized by precise excision, avoidance of local trauma to the tissues and excellent functional control [5-7]. PMID- 9863392 TI - [Indications for carotid endarterectomy in patients with symptoms: when, where?]. AB - Carotid endarterectomy (CEA) is one of the most commonly used surgical methods in the treatment of cerebral stroke with both therapeutic and also prophylactic implications. CEA has been used in surgical practice for 40 years. At the beginning it was very popular and was widely used. Later, the opposite extreme was reached, and its therapeutic efficacy was denied unjustifiably. However, at the beginning of the ninetieth three large controlled studies were completed (North American Symptomatic Carotid Endarterectomy Trial, European Carotid Surgery Trial and Veterans Administrations Symptomatic Trial) and the results of these trials were the basis for establishing the solid criteria for the surgical procedure in some groups of symptomatic patients with stenosis of the internal carotid artery. Thus, CEA was in again. In accordance with the attitudes of the American Association Ad Hoc Committee (1995), evidenced indications for CEA in patients with symptomatic stenosis of the internal carotid artery (in the group with surgical risk less than 6%) include (a) single or recurrent episodes of TIA in the last 6 months, "crescendo" TIA combined with carotid stenosis > 70% with or without plaque ulceration, with or without antiplatelet therapy, and (b) mild stroke in last 6 months with carotid stenosis > 70% with or without plaque ulceration, with or without antiplatelet therapy. The authors report their experience and results of a six-month pilot study of 301 patients, of whom 248 were operated on for symptomatic carotid stenosis with low combined perioperative morbidity and mortality (0.6%). Also, indications for surgical reconstruction of carotid and coronary arteries in patients with marked signs of atherosclerosis in both arterial systems are discussed. PMID- 9863393 TI - [Chronic renal insufficiency: 1) adaptation of nephron function in chronic renal insufficiency and 2) progression of chronic renal insufficiency]. AB - In chronic renal insufficiency resulting from destruction of the vast majority of nephrons, the surviving nephrons adapt their functions to the conditions of vigorous haemodynamic and osmolar overloads. They acquire an appropriate behaviour to preserve the principal renal functions and to achieve the balance of inner space. In the long period of time, similarly as in healthy people. Glomerulotubular balance as well as tubuloglomerular balance distinguish the remaining nephron function, while autoregulation of perfusion pressure along the glomerulus rapidly vanishes. All three regulation mechanisms are characteristic of the nephron function under physiologic conditions. Intense work of the remaining nephrons in chronic renal failure is under the high level controls of the group of hormones, among them are rennin-angiotensin system, arginine vasopressin and atrial natriuretic peptide playing very important and particular roles. Comparison of different published studies emerge the idea that chronically increased arginine-vasopressin levels in chronic renal failure could block the autoregulation of blood flow and hydraulic pressure in glomeruli, which together with other mediator actions give high and fluctuating tense within remaining glomeruli, during every single cardiac cycle. It is probably the main event in the further course of kidney disease progression resulting in definite damage of the overloaded nephrons. Angiotensin II is one of reliably recognised mediators of unfavourable outcome in the process of nephron adaptation in chronic renal failure. Knowing the pathophysiologic processes in the remaining functionally adapted nephrons in chronic renal insufficiency determines a more adequate therapeutic approach in these patients. PMID- 9863395 TI - [Continuous ambulatory peritoneal dialysis in diabetic patients]. AB - Diabetes mellitus has become one of the most prevalent causes of renal disease, and approximately 30% of all insulin-dependent diabetic patients die of renal failure. Renal transplantation is generally the preferred treatment for diabetic patients with end-stage renal disease because it leads to a better quality of life than any other form of dialysis. Because fluid retention, electrolyte and acid-base disturbances are present in diabetics at a higher glomerular filtration rate than in non-diabetics, dialysis is initiated when the creatinine clearance is 10-20 ml/min, levels slightly higher than the recommended 5 ml/min for non diabetics. Since 1978 continuous ambulatory peritoneal dialysis (CAPD) has become the preferred mode of therapy for diabetics. This method of dialysis offers several medical advantages: slow and sustained ultrafiltration, stable cardiovascular status, easier control of hypertension, preservation of residual renal function for a period longer than haemodialysis, steady state biochemical parameters. An additional advantage is a good, tight control of blood sugar achieved by intraperitoneal administration of insulin, which eliminates the need for multiple subcutaneous insulin injections. Intraperitoneally administered insulin closely mimics physiological events, though this route usually requires higher daily insulin doses. Heparinisation and access-related complications, which are the major cause of morbidity while on haemodialysis, are avoided. The social advantages include the possibility of home dialysis, long distance travel, uninterrupted job-related activity. Peritonitis remains the main complication of CAPD in diabetics. The pathogenesis, spectrum of organisms and treatment of peritonitis in diabetics do not differ from those seen in non-diabetics. The technique of catheter insertion, postoperative catheter care and common catheter complications are similar in diabetics to that in nondiabetic patients. Nutritional problems during CAPD may be aggravated by the loss of proteins, amino acids, polypeptides and vitamins in the dialysate. They are especially important in those diabetics who are wasted and malnourished because of poor food intake, vomiting, and intercurrent illnesses. Foot problems are very important in diabetics on CAPD, and a multidisciplinary approach is absolutely crucial. The major contributory factors in the development of foot ulceration are neuropathy, peripheral vascular disease and abnormal stress. With proper selection of patients, diabetics can survive for a long period of time on CAPD. The morbidity and mortality observed during this therapy are primarily related to associated risk factors such as cardiovascular disease, atherosclerotic complications and infections. Certain features of CAPD make it a suitable therapy for diabetics. PMID- 9863394 TI - [Adhesion molecules in kidney diseases (part II)]. AB - Different adhesion molecules are involved in the maintenance of tissue architecture, morphogenesis, immunosurveillance, inflammation, tumour growth, etc. Thus, this review will be directed to the role of cadherins, selectins, integrins and members of the immunoglobuline supergene family in the pathogenesis of glomerulonephritis, acute renal failure, reaction of renal rejection, development of renal tumours, their invasion and metastases. A better understanding of the role of adhesion molecules in nephropathology may provide new aspects of treatment of different forms of renal diseases including tumours. PMID- 9863396 TI - [Aluminum and Alzheimer's disease]. AB - Neuro-toxic effects of aluminium, with disorders mainly in motor coordination, have been proved in epidemiological studies of subjects professionally exposed to aluminium. However, there is, as yet, no adequate evidence that neurotoxicity of aluminium leads to progressive dementia and Alzheimer's disease. It is likely that long-term use of drinking water with a high aluminium concentration, with pH about or less than 7.0, and with low fluoride concentration, is associated with the increased relative risk of Alzheimer's disease. The use of aluminium containing antiacids, antiperspirants and beverages from aluminium cans, are probably of small importance concerning Alzheimer's disease. The relation of aluminium cumulated in the brain to onset of Alzheimer's disease is still unclear, as neuro-pathological lesions, which can be noticed in acute dementia caused with aluminium, significantly differ from those in Alzheimer's disease. Furthermore, morphological and biochemical characteristics of neurofibrillary tangles which occur in the brain after experimental injection of aluminium are different from the tangles in Alzheimer's disease. Results of the studies concerning aluminium concentrations in the brain of patients with Alzheimer's disease are incoherent. To resolve this scientific problem it is necessary to follow-up the prognosis of neurotoxic disorders caused by aluminium. It should be clarified as well whether aluminium in neuro-pathological findings of Alzheimer's disease is an artefact caused by alumino-silicates present in most reagents for tissue-staining. PMID- 9863397 TI - [Modern approach to malignant pleural mesothelioma. 1) Pretherapeutic evaluation]. AB - In this review we outlined the basis of current concepts of pretreatment evaluation in patients with malignant pleural mesothelioma. This tumour, rarely reported until sixties, is mostly connected with exposure to asbestos. Its increased incidence of approximately 50% is noted in last decades. Various histological types of this tumour are well known, but its biology is not well understood. Recent TNM classification and modern diagnostic approach such as computerized tomography and thoracoscopy, as well as standard means of diagnosis are aimed at obtaining early diagnosis and staging in order to undertake the adequate therapy. PMID- 9863399 TI - [Modern aspects of the premenstrual syndrome]. AB - Premenstrual Syndrome (PMS) has being a more and more current problem in gynaecology nowadays. The outstanding of symptoms with their repercussion in the psychosocial sphere, no matter whether the problems of psysiological nature are primary oe secondary characteristics of PMS, all inclusive perception of this syndrome. There are individual various aeriological factors, i.e. pathophysiological mechanisms responsible for PMS development, with precisely known varieties of symptom expression, being dominant and frequent, which contribute to the choice of the most suitable therapy. The paper presents a review of various therapeutical possibilities that will make possible the establishment of up-to-date concepts of the premenstrual syndrome treatment, individualized and harmonized according to the current knowledge of dysfunction of relevant system and presence of neuroendocrinological imbalance, responsible for the development of specific PMS symptoms. PMID- 9863398 TI - [Modern aspects of antiemetic therapy]. AB - We summarized the current knowledge about chemotherapy and radiotherapy-induced nausea and vomiting. Nausea and vomiting are among the most frequent side effects in the treatment of malignancies, and they are very unpleasant for the patient. We reviewed basic aetiological and physiological mechanisms (except that of delayed emesis, which is not enough explored), particularly the role of serotonin in acute chemotherapy and radiotherapy-induced nausea and vomiting. An oncologist cannot make many changes in the treatment of malignancies and patient-related prognostic factors, but he (she) can make changes in the treatment of nausea and vomiting in order to improve the quality of life of patients with malignancies. We also listed some of the most widely used antiemetic drugs with their most important pharmacological properties. Important progress in the control of nausea and vomiting was obtained by the use of selective antagonists of 5-HT3-receptors such as ondansetron, granisetron, tropisetron and dolasetron. Usually ondasetron and granisetron were used. Their clinical activity is similar but better results were obtained with the combination of 5-HT3-antagonists and corticosteroids (complete response was approximately 90%) than by their individual use (complete response was approximately 50%). The problem of delayed emesis has not yet been solved, and best results were obtained with the combination of metoclopramide and corticosteroids. For the control of nausea and vomiting caused by radiotherapy, orally given ondansetron is effective. PMID- 9863401 TI - [The effect of ultrasound in biology]. AB - The article deals with the basic definitions and characteristics of ultrasound. The primary effects of ultrasound such as heat production, microstreaming, cavitation, chemical effects and biologic effects of ultrasound such as teratogeny and mutagenesis of the stronger doses without any clear evidence of deleterious effects, are described. PMID- 9863400 TI - [T lymphocytes in allergic inflammation]. AB - T lymphocytes are the crucial cells in immunopathogenesis of allergic diseases since they regulate the occurrence of allergic sensitisation, synthesis of immunoglobulin E and allergic inflammation. The importance of lymphocyte T is reflected on the fact that after activation by a specific antigen they are able to produce different cytokines responsible for activation and aggregation of specific inflammatory cells in target tissues, promoting the occurrence and maintenance of allergic inflammation. Discovery of functional dichotomy of activated lymphocytes T CD4+ capable of suppressing synthesis of immunoglobulin E (Th1) or stimulate immunoglobulin E and allergic inflammation (Th2) is an important element in elucidation of pathogenesis of allergic inflammation and inadequate synthesis of immunoglobulin E. The immunoglobulin synthesis is regulated by a complex combination of factors and signals where lymphocytes CD4+ play the central regulatory role. PMID- 9863402 TI - [Sodium excretion in children with lithogenic disorders]. AB - INTRODUCTION: The causes of nephrolithisis are multifactorial and have not yet been enough investigated [1]. Hypercalciuria is the most common cause of metabolic nephrolithiasis [2-4]. Close relationship between urinary calcium and urinary sodium has been a subject of reported observations in the past, showing that high urinary sodium is associated with high urinary calcium [5-7]. Hyperoxaluria, hyperuricosuria and cystinuria are also metabolic disorders that can lead to nephrolithiasis. Recent studies have indicated that urinary elimination of cystine is influenced by urinary sodium excretion. Based on these observations it has been hypothesised that patients with high urinary sodium excretion are at high risk of urinary stone disease. The purpose of the study was to investigate sodium excretion in a 24-hour urine and first morning urine collected from children with lithogenic metabolic abnormalities (hypercalciuria, hyperoxaluria, hyperuricosuria, cystinuria), both with nephrolithiasis and without it, in order to determine its significance in urinary calculi formation. PATIENTS AND METHODS: Urinary sodium excretion was investigated in 2 groups of children: patients with lithogenic metabolic abnormalities, but without urinary stone disease (L group) and patients with nephrolithiasis (C group). Both groups were divided into 2 subgroups: patients with hypercalciuria and without it. There were 22 patients in group L (mean age 11.97 +/- 4.13 years), of whom 17 formed a hypercalciuric subgroup and 5 formed a non-hypercalciuric subgroup (3 patients with hyperuricosuria and 2 patients with hyperoxaluria). Group C consisted of 21 patients with nephrolithiasis (mean age 12.67 +/- 3.44 years), of whom 6 formed a hypercalciuric subgroup and 15 formed a non-hypercalciuric group (2 patients with cystinuria and 13 patients without lithogenic metabolic abnormalities). Control group consisted of 42 healthy age-matched children. All subjects had a normal renal function. A detailed history and clinical examination were done, and ultrasonography was performed in all patients. A 24-hour urine, first morning urine and serum specimen were analysed for sodium, potassium, calcium, uric acid, urea and creatinine. Fractional excretion of sodium, as well as urinary sodium to creatinin ratio and urinary sodium to potassium ratio, were calculated from the findings. Sodium and potassium levels were determined by flame photometry, calcium was measured by atomic absorption technique (Beckman Atomic Spectrophotometer, Synchron CX-5 model, USA), uric acid by carbonate method and creatinine by Jaffe technique. Cystine and dibasic amino acids were quantified by ion chromatography. Urinary oxalate excretion was determined by enzyme spectrophotometry. Hypercalciuria was defined by 24-hour calcium excretion greater than 3.5 mg/kg per day and/or calcium to creatinine ratio greater than 0.20 [8]. Uric acid excretion was expressed as uric acid excretion factored for glomerular filtration, according to Stapleton's and Nash's formula [9]. Normal values were lower than 0.57 mg/dl of glomerular filtration rate in 24-hour samples. Mean values were statistically analyzed by Pearson's linear correlation and analysis of variance (ANOVA). RESULTS: Urinary sodium concentration values including urinary sodium to potassium ratios, are shown in Table 1. We found that urinary sodium excretion was significantly increased in patients of both L and C groups when compared with controls (p < 0.05). Further analysis of the subgroups showed that urinary sodium excretion was significantly higher only in patients with hypercalciuria of both L and C groups in comparison to controls (p < 0.05) (Table 2). A significant positive correlation was found between 24-hour urinary sodium to creatinine ratio and urinary calcium to creatinine ratio (r = 0.31; p < 0.001) (Graph 1), as well as between urinary sodium to potassium ratio in 24-hour and first morning urine (r = 0.69; p < 0.001) (Graph 2). (A PMID- 9863403 TI - [Treatment of chronic Wilson's disease in 2 patients using plasmapheresis- clinico-biochemical observations]. AB - INTRODUCTION: The introduction of penicillamine in the treatment of Wilson's disease (hepatolenticular degeneration) was a historical event [1]. D pericillamine (d-PAM) showed some potential side effects such as myasthenia, kidney toxicity, etc. In previous decade the treatment of Wilson's disease (WD) with zinc sulphate started successfully [4]. Danks [7] described the use of exchanged transfusions, peritoneal dialysis and plasmapheresis in the treatment of young patients with WD in acute liver failure. These patients had acute copper poisoning. The results of this study were beneficial. MATERIAL AND METHODS: Therapeutic plasmapheresis (PF) has been used in many diseases in which immunological mechanisms are proved [8, 9]. We started with using PF in the treatment of two young chronic patients with Wilson's disease. The clinical picture of patients became worse, probably due to the decrease in cupriuretic effect of d-PAM. One patient (1) did not take d-PAM regularly. In this study PF was performed with the use of haemonetics V-50 and filter 704. During the treatment with PF, 2000 ml of plasma was always exchanged, i.e. removed. During the treatment with PF the patients were hospitalized at the Department of Neurology and Psychiatry of Children and Young People. CASE REPORT AND RESULTS: Patient No. 1. A 24-year-old man, born in 1965, came with a coarse tremor. He has been diagnosed as WD at the age of 18. Kayser-Fleischer rings were found in the cornea by slit-lamp examination. The disease began when he was 15 years old with polymorphous difficulties. In time tremor became more severe. It was apparent when the patient made any movement. The first treatment with PF (1) lasted from February 13 to March 16, 1989. The patient had 10 PF without any side effect. During the treatment with PF the patient felt better. After PF-tremor was reduced significantly, and subjective and objective condition ameliorated. The patient (1) was readmitted to the hospital on June 24, 1991 for continuation of the treatment with PF. Anamnestic data and neurological examination revealed progression of the disease. His condition became worse, especially tremor. He could not write a single letter (Figures 1-5) and walk without help of the others. His tremor became the severest when he made any kind of voluntary movement. The second treatment with PF started on June 25, 1991 and finished on July 16, 1991. He received 9 PF in this series. The treatment was without side effects. Tremor was reduced approximately by 15%. Plasma copper examined before and after 9 PF showed different values (Table 2). This difference (2.79 mumol/l; 23.70%) in plasma copper level was removed from circulation. Patient No. 2. A 23 year-old man, born in 1966, came to the hospital with acute exacerbation of WD. At the age of 16 latent psychosis was diagnosed. One year later diagnosis of WD was established in the hospital when he was 17 years old. Kayser-Fleischer rings were found in the cornea by slit-lamp examination. The treatment with d-PAM and other drugs (BAL, symptomatic therapies, sedatives, antidepressants, etc.) has been accompanied with good and long-term remissions and short exacerbation. A few years later exacerbation became longer and longer and worse and worse. He was admitted to the hospital on November 28, 1989 with acute relapse of WD. His condition was very difficult, completely bedridden. The treatment with PF started on December 26, 1989 and lasted to January 25, 1990. The treatment with PF was without d-PAM. He was only given symptomatic therapies. After a few PF he demonstrated side effects with nausea, sometimes vomiting, face sweating, pulse rate of about 120/min while blood pressure was normal. Therefore he was given human albumin in the next day, and no side effects were observed. The removal of little plasma copper from blood circulation correlated well with a small improvement in clinical symtpomatology. The rigidity was reduced and voluntary movements bec PMID- 9863404 TI - [Juvenile myoclonic epilepsy]. AB - CONCLUSION: We conclude that despite inevitable variability the clinical picture of JME is characteristic. It is easy to diagnose JME if one thinks of it while the history should be thoroughly analyzed. An EEG recording during sleep confirms the diagnosis. An early diagnosis of JME permits adequate prognosis of the subsequent course of epilepsy, and adequate therapy brings remission in most of the patients. If treatment starts following the large number of severe GTC seizures, the response to therapy is incomplete. The persistency of the illness throughout the life, the need for continuous medication and therapeutic unresponsiveness in cases with late diagnosis, do not justify the increasing misconception that JME is of benign nature. Diagnosis of JME is rare because of insufficient familiarily of physicians with the illness. BACKGROUND: Juvenile myoclonic epilepsy (JME) is an idiopathic generalized epileptic syndrome characterized with the combination of myoclonic, generalized tonic-clonic (GTC) and absence seizures that are readily provoked by sleep deprivation. PATIENTS: Forty-three patients, aged from 14 to 51 years, participated in a 5-year follow up study. Diagnosis was made according to the criteria (Table 1) for diagnosis of JME set by Panayiotopoulos et al. (1994). Nineteen patients made their first contact with a neurologist at the Institute of Neurology and were diagnosed as JME, while the remaining 24 were referred to from other medical institutions with a diagnosis of therapy resistant to focal epilepsy. All patients underwent a somatic and neurological examination, "mini mental test," EEG in waking and CT scan of the brain. Some patients had EEG performed during sleep and some had MRI of the head. RESULTS: JME began between 9 and 26 (average 17) years. All patients had myoclonic seizures, 98% had GTC and 23% absence seizures. The first myoclonic seizure occurred between 9 and 24 years while the frst GTC seizure occurred between 10 and 32 years. Myoclonic seizures (83% of patients) and GTC seizures (70% of patients) occurred most often immediately after awaking. The most frequent provocative factors were insufficient sleep, alcohol abuse and tiredness. Epilepsy in the family was present in 39%, focal neurological deficiency in 9% and pathological findings on of CT and MRI in 7% of patients. Waking EEG was pathological in 77% of patients; it included generalized spike wave discharges in 73%, multiple spike-wave complexes in 33% and focal discharges in 12% of patients, respectively. In all 26 patients tested, sleep EEG was pathological most often with multiple spike-wave complexes in 85% and 3-4 Hz spike-wave complexes in 57% of patients. The correct diagnosis of JME following a comprehensive examination was made in 24 (56%) patients after a delay of 1 to 35 years. In 24 patients with delayed diagnosis of JME the replacement of earlier medication with valproic acid (VPA) induced remission in 18 patients (75%) while 1 patient (4%) experienced a reduction in the number of seizures. Five patients (21%) did not respond to VPA medication: 2 due to a weak compliance, another 2 due to inefficient medication and 1 because of the preexistent malabsorption syndrome. In 19 patients (44%) with initial diagnosis of JME, VPA was introduced immediately upon diagnosis. Of them, 15 (79%) had excellent response to VPA, 1 refused therapy and for 3 patients there is no information. In 2 patients VPA was substituted due to side effects (hepatotoxicity and alopetia) with lamotrigine (low doses), which brought about decrease in frequency and mitigation in myoclonic seizures. PMID- 9863405 TI - [Treatment of aggressive non-Hodgkin's lymphoma with the ProMACE- CytaBOM protocol]. AB - INTRODUCTION: The Working Formulation Classification (for clinical use) divides non-Hodgkin's lymphoma (NHL), according to the nature of the disease and response to therapy into the low, medium and high risk lymphomas. Although these subgroups include different pathohistological types of NHL, they are considered sufficiently homogenous for joint therapy planning [1]. The first generation protocol (CHOP) managed to achieve complete remission (CR) in 50-55% of patients with 30-35% of survival rate [2]. A large four-branch comparative study of SWOG group compared CHOP as the first generation protocol with the third generation protocols ProMACE CytaBOM, m-BACOD and MCOD-D. The results have shown a similar CR and survival rates, so that CHOP is considered a gold standard for the treatment of aggressive NHL [6]. In the light of individual reports stating a high CR rate in the treatment of aggressive NHL by ProMACE CytaBOM [3-5] we present our experience and observations related to the use of this protocol. METHOD: Over the period from 1991 through May 1996 at the Department of Lymphoproliferative Diseases, Institute of Haematology, Clinical Centre of Serbia in Belgrade, we treated 25 patients with pathohistologic evidence of medium to high risk lymphomas, where cases of lymphoblast lymphoma and Burkit's lymphoma were excluded. The median follow-up was 27 months (maximum 63 months). RESULTS: Four of 25 patients were > 60 years. Three of these died. Pathohistological analysis revealed that of 20 cases of medium risk aggressive lymphoma five had diffuse, small cleaved cells, 7 had diffuse mixed and 8 diffuse centroblast cells. Although diffuse NHL with small cleaved cells is classified into clinically indolent lymphomas, two of five patients were in the fourth clinical stage, and three of five patients had a large tumorous mass. In the high risk group five patients had immunoblast lymphoma. Karnofsky index was high in 20/25. According to Ann Arbor criteria 19/25 patients were in IVCS and 7/25 had a large tumour mass. Most patients had clinical symptoms (21/25). Extranodal localization was confirmed in 19 patients. Bone marrow and hepatic infiltrations were most common: 9 and 6 patients, respectively. Eleven patients had a single extranodal localization, while 8 had 2 or more. The median follow-up was 27 months (maximum 63 months), and 21/25 (84%) patients responded to therapy (CR + PR). Complete remission was achieved in 14 patients (56%), and PR in 7 (28%) patients. In the CR group two died, and relapse developed in one after 28 months. In 11 cases CR is maintained. The average duration of CR was 16 months (3-38 months), and PR was maintained for 6 months (20 months in one case). The average survival was 24.5 months (range 3-53). DISCUSSION: The fact that a half of adult patients with disseminated aggressive NHL can be cured with combined chemotherapy is the major oncological achievement in the last 20 years. The protocol combines 4-8 drugs, and the joint report of the SFOG group for lymphoma in over 1200 patients with lymphoma has shown that the second and third generation protocols are not more effective than the standard CHOP or CHOPBleom protocols [6]. The optimum therapeutic protocol in the treatment of aggressive lymphoma is still unpredictable due to the fact that it is inadequate to compare the results of individual institutions with the results of collaborative groups; there is also a significant difference in the prognostic factors in different research groups; there is no sufficient complete and published results that suggest the lower CR than the original reports (which may be related to the evaluation of tumour and remission). There are not sufficient data on the incidence of secondary carcinoma and leukaemia [1]. The decision on the therapy should be based on two lines of information: those related to each particular patient (age, associated diseases) and those related to the tumour (large mass, immunophenotyping, cytoge PMID- 9863406 TI - [Personal experience in embolization of liver hemangiomas]. AB - INTRODUCTION: Cavernous haemangioma is the most often found benign liver tumour. Its size usually does not change, although there are cases in which it grows. Large haemangiomas can cause hepatomegaly, pain in the right subcostal area, and spontaneous ruptures. By modern diagnostic procedures they are detected more often and therefore gained more diagnostic importance. Cavernous haemangiomas, especially giant ones, can be treated surgically (enucleation or resection of a part of the liver), by embolization or by other procedures. The aim of the study was to determine the important role of embolization in the treatment of symptomatic haemangiomas with risk of rupture. MATERIAL AND METHODS: Over a period of 5 years, at the Department of Gastroenterology and Hepatology, haemangioma was discovered in 35 of 178 patients with focal liver lesions. Eighteen (51%) patients were males and 17 (49%) females. In 21 (60%) patients, the size of the tumour was 2-4 cm, in 10 (29%) 5-10 cm, and in 4 more than 10 cm. Ultrasonography, computerized tomography, celiacography, scintigraphy with blood pool and ultrasound guided liver biopsy were used to diagnose haemangiomas. Polyvinyl-alcohol (Ivalon) was used for embolization. Through femoral catheter truncus coeliacus was reached, a. hepatica was catheterized, contrast was injected, and then microembolization of peripheral branches was performed. In 10 patients, because of the size of haemangioma, symptoms or localization, and a high risk of bleeding, embolization was performed. Biochumoral parameters were analyzed on the first, the second and the seventh day after the intervention. Within the period of five years, control ultrasound examinations were performed in all patients, and results were compared. In 9 patients control liver scintigraphy with blood pool was carried out. RESULTS: Embolization was performed with polyvinyl-alcohol. During angiography which followed, avascular zones were seen. There was no statistically significant difference between biochumoral parameters before and after embolization. Five years after the embolization, a reduced size of haemangioma was found in 8 patients. The echosonographic appearance of the tumour was changed in almost all patients. All clinical symptoms disappeared. There was no bleeding. In 8 of 9 patients liver scintigraphy with blood pool was performed, and there were no "warm fields." DISCUSSION: Due to modern diagnostic procedures, haemangiomas are now more often detected. However, ultrasonography is not always specific in discovering haemangiomas. Liver scintigraphy does not always reveal the typical shape of these tumours. Every procedure has its advantages and disadvantages. Once haemangioma is detected, it is the question how to treat it. Experience of most hepatologists suggests that interventions should be performed only in case of symptomatic haemangiomas, progressively growing haemangiomas, and in case of the high risk of bleeding. Embolization of the hepatic artery, previously used only as the first part of surgical procedures is now used as the only procedure in the treatment of these tumours. Some authors reported pain and fever after this intervention, which were also noticed in our patients. The reported agranulomatous arteritis with eosinophilic infiltration was not found in our patients. There were no significant changes in biochumoral analysis; this finding confirmed that there was no necrosis around embolized haemangioma. On the basis of the follow-up of our patients we came to the conclusion that embolization of haemangioma, performed by an experienced radiologist, is a very useful procedure in the therapy of symptomatic haemangiomas and haemangiomas with a high risk of bleeding. PMID- 9863407 TI - [Chronotherapy with high dose carboplatin, 5-fluorouracil and leucovorin in advanced colorectal carcinoma]. AB - INTRODUCTION: Fluorouracil (5-FU) has remained the mainstay of treatment of advanced colorectal cancer disease for nearly 40 years, and despite the implementation of various strategies to increase response rates no substantial improvement in survival has been achieved. 5-FU efficacy has been enhanced by modulating its cytotoxicity with leucovorin (LV) or by administering it as a continuous intravenous infusion. These regimens resulted in a few-fold improvement of tumour response rate in patients with metastatic disease compared with standard 5-FU treatment. The figures are still low (25%), and survival is affected only modestly [1, 2]. Cisplatin and carboplatin are also able to modulate 5-FU cytotoxicity in experimental systems [3]. Recent results of experimental studies suggested that high doses of carboplatin were necessary to achieve biochemical modulation of 5-FU cytotoxicity in vivo [6]. The use of chronobiologically determined drug over a 24-hour period may allow an increase in antitumour effect and a decrease in side effects. Experiments in animals revealed large changes in the toxic effects of 5-FU and platinum analogues, depending on the circadian rhythm of drug administration [9, 10]. The aim of the study was to investigate the toxicity and efficacy of combination of high dose carboplatin, fluorouracil and leucovorin in patients with advanced colorectal adenocarcinoma. Carboplatin and 5-FU were administered in circadian-dependent rhythm regimen in a 4-h infusion. PATIENTS AND METHODS: An open, prospective study was carried out in 99 patients enrolled in the study. The following treatment schedule was used: carboplatin 150 mg, a 4-hour-infusion, start at 8 a.m., day 1-7; 5-FU 750 mg/m2, a 4-hour-infusion, start at 6 p.m., day 1-5; leucovorin 100 mg/m2, bolus i.v. at 8 p.m., day 1-5. The cycles were repeated every 28 days. Dose modifications were not planned. Treatment response and toxicity were evaluated according to WHO criteria [11]. Time to progression was calculated from the beginning of chemotherapy. Survival was calculated from start of chemotherapy using the Kaplan Meier method [12]. RESULTS: We treated 99 patients (27 colon and 72 rectal cancers) with metastatic diseases. There were 52 males and 47 females, average age 58 years (range 34-72). There were 82 patients (pts) with previous surgical treatment, 30 pts. with previous radiotherapy and 19 pts. with previous adjuvant chemotherapy. Performance status was 0 for 9 pts., 1 for 53 pts., 2 for 33 pts. and 3 for 4 pts. We carried out 469 cycles (mean 4/pts.). Ninety seven pts. were evaluated. The response rate was 28% (95% CI: 19.9-37.9) with 4/97CR, 23/97PR, 48/975D and 22/97PD. Mean time to progression was 5.5 months (range 2-18). The mean survival time was 7 months (range 4-27). The mean survival time was 11 months for responders and 6 months for non-responders. WHO grades 3 and 4 leukopenia were observed in 19 pts (19.2%). WHO grades 3 and 4 thrombocytopenia occurred in 21 (21.2%) pts. The main nonhaematologic toxicity were diarrhea, nausea/vomiting and mucositis. No grades 4 of nonhaematological toxicity were seen, except diarrhea grade 4 was observed in one patient. There was no life threatening toxicity. DISCUSSION: Carboplatin is able to modulate 5-FU cytotoxicities [3]. A high dose of carboplatin is necessary to achieve biochemical modulation of 5-FU cytotoxicities in vivo [6]. The studies of colon tumour cell lines demonstrated sensitivities to carboplatin when used at clinically achievable dose level [4]. Clinical studies of carboplatin, as a single agent or in combination, in the treatment of colorectal cancer, reported controversial results [19, 20]. Animal studies indicated large and predictable changes in the toxic effects of carboplatin and 5-FU, depending on the circadian rhythm of drug administration. Mechanisms included 24 h changes in the activities of several enzymes involved in 5-FU and carboplatin catabolism or in the anabolism of its cyt PMID- 9863408 TI - [Specific skin reactions induced by individual pollen preparations in hypersensitivity persons]. AB - INTRODUCTION: Skin tests are the most useful single modality for demonstrating an IgE-mediated mechanism underlying clinical symptoms. Skin reactivity to an allergen depends on person's exposure and genetic factors influencing the IgE response. However, the reproducibility of allergy skin tests has been shown to be variable depending, among other things, on the allergen extract employed and the kind of technique used. These variations have led to controversy regarding clinical relevancy of allergy skin testing [1-7]. We examined the relationship of skin-prick test and serum allergen-specific IgE in pollen susceptible adult persons. We estimated changes in the immediate skin reactivity to pollen allergens by testing patients on more than one occasion: a) during and out of pollen season; b) before and after completing specific immunotherapy, and c) who had been tested with diverse concentrations of the same pollen-allergen. We correlated the degree of skin reactivity at the initial and subsequent testings with the aim of estimate diagnostic relevancy of such performed allergy skin testings. MATERIAL AND METHODS: Study was performed in 64 persons (aged from 17 to 43 years) suffering from rhinitis or rhinoconjunctivitis or asthma. They were diagnosed for the first time [8, 9]. None of selected persons had proven dermographism [1, 9]. Study protocol Skin prick tests were performed according to recommended protocol reviewed elsewhere [9]. Blood samples for total and allergen specific IgE were taken in the morning on the test-day and sera were stored until analyses. Testing was performed with in house standardized extract of grass pollens (each of 5000 AU/allergy unit per milliliter): Dactylis glomerata and Phleum pratense and Lolium perenne (produced by Torlak Institute, Belgrade). Testing was performed in selected groups of patients: a) during and out of pollen season; b) before and after allergen-specific immunotherapy, and c) with three diverse concentrations of Phleum pratense allergenic extract (5000 AU, 7500 AU and 10,000 AU/ml). During the study a medication was not allowed at least 7 days prior to the skin testing (14 days for hydroxyzine). Alergen-specific IgE in serum (RAST) Determination of serum allergen-specific IgE was performed by using commercially available Pharmacia kit (EIA RAST Phadesim). For allergen-specific IgE results were expressed in classes (1 minimal to 4-maximal concentrations) determined by manufacturer. Study was performed under approval of the Ethic Committee of our hospital. For statistical analysis we used t-test and test of variance (ANOVA). Results were considered significant if p < 0.05. RESULTS: Influence of allergen-specific serum IgE concentration on local skin reactivity was estimated in 23 persons (17 females, 6 males), and is shown in Table 1. The largest papule diameter was registered in high susceptible persons (RAST class 4). Influences of season when testing was performed and specific immunotherapy were estimated in 26 persons (19 females, 7 males) (Figures 1 and 2). The effect of diverse allergenic extract concentrations was estimated in 15 adults (9 females, 6 males, mean age 26.4 years) and results are shown in Figure 3. DISCUSSION: In the study it has been shown that serum concentration of allergen specific IgE positively correlated with the skin reactivity to pollen-extract. In addition, allergen specific immunotherapy as well as the concentrations of the used allergenic extract influenced significantly local skin reactivity. On the contrary, season when testing had no influence. According to our results, it seems that skin reactions (papule), equal or larger than 6 mm in diameter, could be of diagnostic value concerning pollen-monotest application (Table 1). Meanwhile, with significance of 95% we could postulate that in persons tested with the lowest concentration of allergen-solution (5000 AU/ml), skin prick reactivity defined by the papule of 4 mm or larger can replace in vitro PMID- 9863409 TI - [Doppler-duplex ultrasonography in the diagnosis of cavernous portal vein]. AB - Prehepatic portal hypertension caused by cavernous transformation of the portal vein has been more and more considered as a multiorgan disease with circulatory changes in numerous organs related to systemic and splanchnic vascular network [1]. Honeycomb-like, spongy, cavernous portal vein is a rare clinical and pathoanatomical entity which usually results from portal vein thrombosis. Recanalization and neovascularization processes lead to cavernomatous transformation of the portal vein lumen and formation of periportal collateral hepatopetal venous varices (Petren's veins) [5, 6]. Recently, with Doppler ultrasonography and angiography cavernous portal vein has been identified as the cause of prehepatic portal hypertension. Usage of color Doppler and duplex Doppler ultrasonography has greatly contributed to diagnostic efficiency, while therapeutically, the disease remains a serious and controversial problem. METHODS: At the Institute of Digestive Diseases, Clinical Centre of Serbia, 8 patients with cavernous portal vein were studied in the period 1995-1997. Real time duplex and color Doppler ultrasonography (Toshiba-SSA 100A with sector duplex probe 3.75 MHz, and 9 ATL with color Doppler convex duplex probe 3.5 MHz) were used. Indirect (arterial) portography was used for imaging of lienoportal system in the venous phase of angiography as follows: catheterization (Seldinger's technique) of the coeliac trunk or lienal artery, and catheterization of the superior mesenteric artery. Indirect portography was performed by injection of 60-80 ml of the contrast medium by an automatic pump, at 10-14 ml/sec, i.e. 8-10 ml/sec by the digital technique [7]. Peroral fiberendoscopy was performed in all patients by Olympus GIF-XQ 10 endoscope. RESULTS: In our study the conventional ultrasonographic examination failed to provide an appropriate image of the normal portal vein. In hepatoduodenal ligament multiple tubular and round structures were seen, revealing an atypical honeycomb or spongycavernous shape of the venous lumen (Figs. 1 and 2). Doppler ultrasonography of the lumen of these venous collateral structures revealed a continuous, hypokinetic flow, mid-rate 7.4 cm/sec, which was always hepatopetally directed (to the liver). Color Doppler ultrasonography detected extensive portosystemic collateralls in all patients, and varices in the gallbladder wall in 1 patient. The results of indirect portography correlated well with Doppler ultrasonographic findings. In all patients hepatopetal flow was found (Figs. 3 and 4). The aetiology was diverse: idiopathic, liver cirrhosis, haematological diseases, Crohn's disease and Marfan's syndrome. Two patients had IV degree varices in the distal third of the oesophagus, and 4 patients had II/III degree varices. Patients with posthepatic liver cirrhosis and Crohn's disease had no varices in the distal third of the oesophagus and gastric fornix. DISCUSSION: Since Pick (1909) described this malformation as the hepatopetal collateral, the haemodynamic concept of this entity has not been changed. Doppler ultrasonography and angiography confirm that the blood flow in cavernomas is hepatopetal, i.e. compensated and functional. Cavernous transformation of the portal vein is clinically manifested by bleeding from oesophagogastric varices. Haemathemesis is the most alarming complication and may be the first clinical sign. The haemorrhage is usually recurrent and profuse, but in most cases it is tolerated well owing to preserved hepatic function in patients without liver cirrhosis [19]. Portosystemic collateral circulation may take place via retroperitoneal and other spontaneous venous shunts, not involving the left gastric vein or vv. gastricae breves, when oesophagogastric varices are absent (our patient with Crohn's disease and posthepatitic B cirrhosis). Splenomegaly with hypersplenism is always present with cavernous transformation of the portal vein, and usually precedes the occurrence of gastrointestinal hae PMID- 9863410 TI - [Effect of various methods of treatment in chronic renal insufficiency on the quality of life in patients]. AB - Interest in measuring the quality of life (QL) in relation to health care has increased enormously in recent years. This is also true for end-stage renal failure where it is important not only to provide a better survival but also the quality of that survival. The aim of this study was to assess the relative influence of different kinds of treatment on end-stage renal disease after the patients' evaluation of their overall QL. We studied 167 patients receiving conservative treatment (45), haemodialysis (44), haemodialysis and erythropoieth (36), and continuous ambulatory peritoneal dialysis (42). The patients completed an original questionnaire consisting of 37 questions divided in five groups and generating 15 QL variables: personal data (name, gender, age, basic kidney disease); sociodemographic data influenced by the illness (family history, working ability, employment status); general health characteristics (fatigue, appetite, wound healing, sleep, resistance to cold); aspects of private life that are mostly influenced by the disease (social interaction, traveling, mood, sports, sexual life), and patients subjective assessment of their condition (self care and happiness). Patients on haemodialysis showed lower levels of QL than that on peritoneal dialysis related to fatigue (p < 0.01), working ability (p < 0.05), wound healing (p < 0.05), and appetite (p < 0.01) compared to the conservative treatment. Peritoneal dialysis had also a statistically significant positive influence on fatigue (p < 0.05) compared to conservative treatment. However, erythropoletin treatment showed better results with regard to traveling (p < 0.05), resistance to cold (p < 0.01), self care (p < 0.05) and mood (p < 0.05) compared to peritoneal dialysis, and working ability (p < 0.05), fatigue (p < 0.05) and mood (p < 0.05) compared to conservative treatment and haemodialysis. PMID- 9863411 TI - [Phonosurgical treatment of vocal cord cysts]. AB - INTRODUCTION: Functional-traumatic lesions of the vocal folds include mucous stranding, "nodular" lesions, polyps, cysts, contact hyperplasia and haematoma [1]. They all appear at the predilection sites of the vocal fold, at the junction between the anterior and middle thirds, slightly below the free edge. Vocal fold cysts are also located at the predilection site. They interfere with the glottic closure and vibration process. The treatment consists of surgical enucleation. This paper is aimed to present our results with direct and indirect surgical procedures. MATERIAL AND METHOD: A series of 63 vocal fold cysts was operated by the use of direct microlaryngoscopy (DML), indirect microlaryngoscopy (IMS) and videostroboscopy (IVCS). RESULTS: Over a 10-year period 1550 surgical procedures were performed for benign lesions of the vocal folds, including 63 cases of vocal fold cysts (4.1%). A very satisfactory phonusurgical result was obtained. Recovery of the vibration pattern was after DML procedures within 3 weeks in 66.6% of patients, while it was apparently faster after indirect operations. The latter were performed only for minor cystic lesions, and therefore direct and indirect procedures cannot be compared to vibration recovery. DISCUSSION: A careful surgical manipulation is required for the enucleation of vocal fold cysts, sparing the local tissue. DML is used in the majority of cases, while indirect procedures can be used only in minor lesions, where enucleation is not possible. PMID- 9863413 TI - [The role of prostaglandins in allergic inflammation]. AB - Prostaglandins likewise leukotriens are proinflammatory mediators resulting from metabolic degradation of the arachidonic acid originating from membrane phospholipids. The most important products of enzyme cyclooxygenation of arachidonic acid are prostaglandins D2, E2, F2a, tromboxane A2 and prostacyclin. Prostaglandins express their tissue effects via the five basic receptor types. Within the allergic inflammation activated mast cell synthesizes prostaglandin D2 (first lipid mediator) which has bronchoconstrictive and vasodilating effects and attracts neutrophilic leukocytes. Moreover, it also participates in the late phase reactions, six hours subsequent to the exposure to the allergen. This mediator is also important in pathogenesis of urticaria, allergic rhinitis and allergic bronchial asthma. In addition to prostaglandin D2, prostaglandin F2a and tromboxane A2 also have bronchoconstrictive actions, while prostacyclin and prostaglandin E have bronchodilating effects. Inhalation of prostaglandin E prevents asthmatic attacks caused by allergens, strain, metabisulfite and ameliorates attacks of aspirin asthma, which confirms the hypothesis that aspirin asthma is based on cyclooxigenase inhibition and increased leukotriene production. In patients with atopic dermatitis, prostaglandin E has suppressive effects on Interferon gamma production by Th1 helper cells and increases production of Interleukin 4 by the Th2 cells. Tromboxane A2 plays a certain role in the development of bronchial hyperreactivity and late asthmatic response. Prostaglandins are also important mediators in the pathogenesis of allergic conjunctivitis. Most of nonsteroidal antiinflammatory drugs inhibit the enzyme cyclooxygenase and thus also prostaglandin biosynthesis and release. PMID- 9863412 TI - [Leukotrienes in allergic inflammatory reactions]. AB - Production of leukotrienes, lipooxygenase products of arachidonic acid metabolism, plays an important role in inflammatory reactions, particularly well studied in bronchial asthma. Lipooxigenase-5 and lipooxygenase-activating protein 5 are crucial in the production of leukotrienes with potent biological activities. Leukotriene B4 is a leukocytic chemoattractant and it induces aggregation and adherence of leukocytes to endothelial vasculature. Sulfidopeptid leukotrienes (C4, D4 and E4) are potent bronchoconstrictors, producing mucous secretion in the airways and increasing vascular permeability. Leukotrienes participate in the process of inflammation, as well as in early and late asthmatic responses. They are found in the blood, liquid obtained upon bronchoalveolar lavage as well as in the urine, irrespectively whether bronchospasm developed spontaneously or it was induced by an allergen. Administration of the specific leukotriene receptor antagonists or leukotriene synthesis inhibitors ameliorates the symptoms and signs of bronchial asthma. PMID- 9863414 TI - [Functional evaluation in patients with kidney calculi]. AB - Nephrolithiasis is a common disorder and a significant problem because of incidence, recurrence and severe consequences. Stone disease is a surgical as well as a medical problem. Major progress has been made recently in understanding the pathophysiological disturbances responsible for stone formation as well as in the techniques of stone removal. The introduction of extracorporeal shock wave lithotripsy has considerably reduced the need for surgery. Improvements in methods of kidney stone removal have not diminished the need for the application of an effective prophylactic program. The internist should take a complete history of stone events (number, composition, location and outcome of stone event), family history of stones, dietary habits (focusing on the consumption of animal protein, salt and dairy products), medications and physical examination. Radiopaque stones should be documented by plane X-ray films. Ultrasonography should be used to image calculi that are nonopaque, and to easily distinguish them from masses such as tumour or blood clot. Computed tomography is also an excellent method for imaging nonopaque renal calculi but higher cost and radiation exposure are disadvantages [2]. Crystallographic analysis is the essential diagnostic procedure. If available, previous stones should also be examined. "In stone disease, everything is measurement. What the laboratory cannot tell you, you will not know; what it tells you in error, you will not correct by using your instincts, your medical experience, or your art [3]". Reliable diagnostic protocols are available for the identification of different causes of stones. The complexity of protocols depend on the severity of nephrolithiasis. Patients with a single stone episode undergo simple protocol, and extensive detailed protocol is used for patients with recurrent stone disease, or patients at increased risk. Simple protocol, besides the already mentioned history of stone events, radiographic investigation and crystallographic analysis, includes serum urea, creatinine, uric acid, sodium, calcium, phosphorus and protein levels, urinary pH and volume, urine samples for culture and urinary calcium, uric acid, oxalate and citrate. Extensive metabolic evaluation includes simple protocol, determination of serum levels of alkaline phosphatase, parathyroid hormone, thyroxin, magnesium. A 24-h collection of urine specimen is analysed for urea, creatinine, uric acid, calcium, phosphate, sodium, magnesium, oxalate and citrate. Extensive protocol includes specialized evaluation tests [5]. Urinary acidification test is important for detecting distal renal tubular acidosis. Two 24-h urine specimens are collected while the patient is on the regular diet. The patient is then placed on a restricted diet (400 mg of calcium and 100 mEq of sodium) for a week, and another 24-h urine sample is collected. After that fasting and calcium load tests are performed (Sheme 1). Fasting urinary calcium is used to detect renal calcium leak, and calciuric response to oral calcium load provides an indirect measure of intestinal calcium absorption. Diagnostic criteria for major forms of stone disease [8] are presented in Table 1. There are some still unsolved questions: does time after passage of stones or urological intervention influence the frequency of urine abnormalities that can be detected; are there differences in 24-h urine composition between weekdays and weekends: what is the prevalence of the most important urinary risk factors of recurrent idiopathic calcium nephrolithiasis: do male patients differ from females with respect to urinary risk factors or recurrent idiopathic calcium nephrolithiasis? [7]. PMID- 9863415 TI - [Importance of use of ultrasound in the early diagnosis and treatment of hip joint developmental disorders]. AB - Developmental dysplasia of the hip has been a problem for over a thousand years. Clinical signs are insufficient for an exact diagnosis, X-ray diagnosis is reliable only at the age of six months and later. Advantages of ultrasound diagnosis (used immediately after birth), classification of findings and basic treatment are described. PMID- 9863416 TI - [125 years' of the Serbian Medical Society]. AB - In the second half of the last century and under the influence of the European civilization, Serbia abandoned the conservative and patriarchal way of life and began to introduce a new, contemporary political, cultural and social spirit into the country. The development of these civilizing features was under the influence of young intelectuals who, as former scholarship holders of the Serbian government, were educated in many European countries. Among them, there was a group of physicians who returned to the country after having completed their education. They were carriers and holders of the contemporary medical science in Serbia and the neighbouring areas. On April 22, 1872 a group of 15 physicians founded the Serbian Medical Society with the intention to offer an organized medical help and care to the population. The first president was Dr. Acim Medovic and the first secretary Dr. Vladan Dordevic. At the meeting held on May 15, 1872 the text of the Statute of the Society was accepted and immediately submitted for approval to the Ministry of Internal Affairs. In the letter addressed to the minister of internal affairs the following reasons were cited: "... The Belgrade physicians feeling a need for having the main office for their professional and scientific meetings, for which they will find the opportunity and the funds, and in spite of their hard medical labor which requires almost all their time, decided to establish the Serbian Medical Society because they wish to be in trend and follow-up the medical progress and exchange the latest medical information not only among them but also with other graduated doctors living in areas with the Serblan population as well as with all scientists who are willing to contribute to the development of medical science in Serbia...". In the first year of its existence the Serbian Medical Society had 9 regular members, 1 honorary member and 34 corresponding members from Serbia, Slavic and other foreign countries. On August 5, 1872 it was decided to start the publication of a professional journal "Srpski arhiv za celokupno lekarstvo" (Serbian Archives of General Medicine). The journal has still been edited. On suggestion of the Serbian Medical Society the Law of Health Care was promulgated in 1881. It was translated into German and French languages and sent to about 400 addresses in different European countries with the request for their opinion and suggestion. The reply of the Vienna Medical Society was as follows: "... While the Austrians carry out some stupid regulations of health care, at the same time a small Balkan country, Serbia promulgated a Law according to which no one, including the King, the Government or a political party dare not use a cent intended for health service, treatment of the sick people and payment of physicians...". On the occasion of the centenary of the First Serbian insurrection and coronation ceremonies of the King Petar I Karadjordjevic, and under the King's patronage, the First Congress of Serbian physicians and naturalists was organized from 5 to 7 September 1904. There were 433 participants of whom more than 100 foreign participants. A Serbian professor of infectious diseases at the University School of Medicine in Vienna, Dr. Jovan Cokor, presented a paper with results of his studies of tuberculosis according to which tuberculosis could be transmitted from a sick cow to man; in this way he complemented the results and explanations of Dr. Robert Koch who discovered Mycobacterium tuberculosis. In 1907 the Serbian Medical Society organized in Belgrade the First meeting of Yugoslav surgeons. At that time, the foundation of a University school of medicine was planned. The Serbian Red Cross Organization was initiated by the Serbian Medical Society in 1876. On February 2, 1891 a procedure was brought for the establishment of the Medical Chamber. Its activity began in 1901. During the First and Second world wars the activity of the Serbian Medical Society was di PMID- 9863417 TI - [Anniversaries of the Serbian Medical Society. 1947. 75 years since the founding of the Society]. AB - During the Second World War the country was again occupied by the enemy, and the Allies were liberating it--but it was bombarded by both! Graves and tombs covered the country once again--once again Serbian people had to suffer the greatest possible sacrifies as an ally with the progressive part of the world. Serbian medical association (SMA) had to restrain its activities once again in 1941 and share the fate of its people. Since the country was occupied the University of Belgrade and Medical school were closed. SMA restored its activities in 1946 and since then it turned to creating a new organization--establishing local branches and specialist groups all over the country. The Association initiated activities on improving professional skills of its members in order to fulfil its goals: solving the most important health care problems "which have flooded the country after the occupation of the country and plight of the people". As soon as the first post-war conference in the Autumn of 1946 took place, the Association brought resolutions concerning the "question of tuberculosis, question of paediatrics and question of dentistry in attempts to protect the people from the social disaster (tuberculosis)" and to deal adequately with the problem of health care of the young, as the most important task of the health care policy," and also to establish a "University School of Dentistry as soon as possible, in order to provide a sufficient number of dentists". The established tasks are being fulfilled, local branches and specialist groups are formed, thanks to commitment of the members of the SMA board, and above all because of the "hearty response of the Medical school colleagues--to go to all parts of the country"--as it was stated in the annals of the SMA. Engaged entirely in organizing its activities and professional improvement of its members, the SMA was most widely involved in dealing with basic problems of health care of the population. Therefore, the 75th Jubilee of the SMA is a crucial part of its history and a period of great success in creating a health-care system which brought our country among the most advanced in the world, regarding the accessibility of health care to the population! PMID- 9863418 TI - Odontoblast differentiation: a response to environmental calcium? AB - The response of the dental pulp to calcium hydroxide has been well described but the process of pulpal repair leading to dentinal bridge formation appears complex and the mechanisms remain incompletely understood. Through the precise regulation of the free calcium ion in the cytosol, cells have been able to utilize anions such as phosphates for a wide range of activities such as energy production (oxidative phosphorylation). As anions are abundant in the cytosol, intracellular levels of calcium ions are kept low, several orders of magnitude less than that of the surrounding extracellular matrix. Consequently, cells are able to use calcium ions for the regulation of many cellular events. The binding of extracellular molecules such as cytokines, hormones or antibodies, with receptors on the plasma membrane may result in short- or long-term modifications to cellular metabolism, including the mechanisms of intracellular calcium homeostasis. Cell survival depends upon the ability to adapt to changes in the cell's micro-environment. Adaptation in turn results in altered cellular activity that may be interpreted as showing that the cell has become more or less specialised. In some instances this may include the resumption of mitotic activity. If the rate or magnitude of change exceeds a cell's adaptive capacity, the cell dies. Responses of cells to alterations in their environment are reviewed as they may provide an explanation for the success of calcium hydroxide in facilitating pulpal repair and the differentiation of odontoblasts. PMID- 9863419 TI - A slow release calcium delivery system for the study of reparative dentine formation. AB - Several liquid, semi-solid and solid delivery systems were formulated and tested to devise a method of reproducibly administering accurate micro-doses of calcium into a 700 microns diameter cavity in a rat maxillary incisor tooth, in the absence of hydroxyl ions. Development of this delivery system was necessary to facilitate studies of the mechanisms of pulpal repair and odontoblast differentiation. The principal requirements for the delivery system were that it should be easily administered into a small pulp exposure in the rat incisor and that a greater than 1000-fold range in calcium ion concentrations could be incorporated and delivered for a period of 2-3 days, preferably in an acidic environment to minimize the effect of non-specific nucleation under alkaline conditions. Poly- (ethylene) glycol microspheres were found to be an ideal vehicle. Under the in vitro dissolution conditions used, complete release of all calcium salts occurred within 12-15 hours, except for the very water-insoluble calcium stearate. It was anticipated that the release of calcium ions would be significantly more prolonged in vivo because of the physical constraints of the prepared cavity as well as the restricted access to fluid flow. PMID- 9863420 TI - Adhesive bonding of fragmented anterior teeth. AB - This in vitro study investigated the fracture strengths of fractured incisors after reattachment to the remaining tooth structure, using either Gluma dentin bonding agent together with 1 of 13 different experimental resins or 1 of the following dentin-bonding agents: Gluma+ (experimental modification of Gluma), Panavia21, ScotchBond1 (SingleBond), All-Bond2, Dentastic, SuperBond, Prime&Bond 2.1. No significant differences were found in the experiments with various experimental resins. The fracture strength of teeth rebonded with Gluma, All Bond2, Dentastic, SuperBond or Prime&Bond 2.1 showed mean fracture strengths in the range of 8.7-11.2 MPa, which were significantly different in all cases from that of intact teeth (16.3 MPa). Teeth bonded with Gluma+, Panavia21 or ScotchBond1 (SingleBond) showed mean fracture strengths of 15.5, 14.6, and 14.2 MPa, respectively. These fracture strengths were not significantly different from that of intact teeth. PMID- 9863421 TI - Effectiveness of four chemical solutions in eliminating Bacillus subtilis spores on gutta-percha cones. AB - Gutta-percha cones should be free of pathogenic micro-organisms before being used for root canal filling. This study was carried out to evaluate the effectiveness of four chemical agents in eliminating Bacillus subtilis spores from gutta-percha cones. The solutions tested were 5.25% sodium hypochlorite, 2% glutaraldehyde, 2% chlorhexidine digluconate, and 70% ethyl alcohol. The gutta-percha cones coated with spores were placed into contact with the chemical agents for 1, 3, 5 and 10 min. The results showed that 5.25% sodium hypochlorite was effective in destroying the spores after 1 min of contact. Glutaraldehyde, chlorhexidine and ethyl alcohol did not decontaminate the gutta-percha cones even after 10 min of contact. PMID- 9863422 TI - The effect of systemic tetracycline on resorption of dried replanted dogs' teeth. AB - The use of systemic penicillin after an avulsion injury has been recommended to decrease the occurrence of resorption complications. Tetracycline antibiotics have been reported to possess anti-resorptive properties in addition to their anti-microbial actions. The purpose of this study was to evaluate histologically the effect of systemically administered tetracycline and amoxicillin on the inhibition of resorption due to attachment damage after replantation of dogs' teeth. Thirty-one roots from the teeth of four beagle dogs were endodontically treated to inhibit subsequent inflammatory root resorption of pulpal origin. They were then extracted and left to bench dry for 1 h to ensure severe periodontal ligament damage before replantation. For the teeth in the experimental groups, two dogs were given tetracycline hydrochloride and the other two dogs were given amoxicillin, administered orally, on the day of extraction/replantation and for the following 6 days. The control group were teeth in these animals treated in the same manner but where no antibiotics had been given. After 12-16 weeks, the dogs were sacrificed and histological sections were prepared and evaluated for complete healing, inflammatory and replacement root resorption, and the groups were compared. It was shown that healing in the amoxicillin and the control groups was poor (10.90% and 11.28%, respectively), while for the tetracycline group, 35.45% showed complete healing. Individual teeth with over 50% complete healing sites were considered as having good healing, and significantly more of these teeth were found in the tetracycline group (5 of 11 teeth) compared to the amoxicillin (1 of 11 teeth) or control group (1 of 8 teeth). PMID- 9863423 TI - Radiographic assessment of simulated external root resorption cavities in maxillary incisors. AB - Upper central and lateral incisors of three human skulls were used to assess radiographically the possibility of detecting cavities of different sizes and locations, drilled to simulate external root resorptions. The sequence of radiographs was evaluated by three endodontists, who were unaware of the experiment being done, using a 2 x magnifier. The first observation was done without comparing the radiographic images of the different cavities with the preoperative radiograph. For the second round of observation, each sequence was compared with the preoperative X-ray. The results showed that small cavities were more difficult to detect than the medium and large ones. Moreover, cavities located on the proximal surfaces were more easily detected than those located on the buccal surfaces. Finally, when the observers could compare with the preoperative radiographs (second round of observation), the rate of detection was considerably higher. PMID- 9863424 TI - Experimental actinomycosis in mice induced by alginate gel particles containing Actinomyces israelii. AB - The present study aimed to make an animal model for investigating chronic infection. Bacterial cells of Actinomyces israelii (ATCC 10048) were packed in alginate gel particles and injected intra-peritoneally into BALB/c mice. Actinomycotic lesions were induced efficiently in 9 mice out of 12 after 3 or 9 weeks. Actinomycotic lesions were not produced by injecting a bacterial suspension of A. israelii except in one animal. Neither did injection of gel particles without bacteria induce lesions. Bacteria survived in the lesions for at least 9 weeks after the injection, and serum IgG levels against the bacteria were significantly elevated in the animals, indicating that the bacteria were protected from immunological elimination and activated humoral immunity in the animals. Histopathological observation of the lesion by staining revealed that the bacteria in the lesions were acid-fast and seemed to become resistant to phagocytosis. The bacterial masses were surrounded by inflammatory cells, including polymorphonuclear leukocytes and macrophages, and were separated from the host cells by a fringe-like structure similar to the capsular structure of natural sulfur granules. The present study indicated that the use of an alginate gel was effective in inducing actinomycotic lesions in mice. PMID- 9863425 TI - Root resorption of replanted teeth: an SEM study. AB - Two replanted and subsequently expelled upper central incisors were observed under a scanning electron microscope to study the changes brought about by root resorption. The severity of resorption and the distribution of the affected areas were different in the two specimens. In the first case, large, deep excavations were found along a continuous bone defect which exposed the opposite root canal wall. Bacteria-like elements were found on the bottom of the eroded areas both on the outer and on the inner root surfaces. In the second case, resorption denuded the apical portion of the metal post. Erosive lacunae were visible in some areas while tubiform excavations were present close to the apical margin exposing the collagen network. The roots in the first case likely underwent inflammatory resorption while replacement resorption was thought to have occurred in the second case. PMID- 9863426 TI - Craniomandibular/temporomandibular/cervical implications of a forced hyper extension/hyper-flexion episode (i.e., whiplash). AB - Clinicians now appreciate the full chain of tissue damage of the interconnecting muscles, tendons, ligaments and fascia: this compromised linkage from the skull through the suboccipital musculature to the cervical spine and anterior/posterior cervical muscles, from the check-rein ligaments and muscles extending from the skull and maxilla to the mandible, from the suprahyoid musculature connecting the mandible through to the hyoid bone, from the hyoid through the infrahyoid to the supporting shoulder girdle, all contribute to damaged interconnecting matrices. Consequently, unresolved tissue damage in any of this linkage becomes mutually provocative during function to any part of the linkage. Diagnostics, therefore, must necessarily include examination of this total linkage; similarly, eventual treatment protocols must resolve tissue damage in all of this interconnecting linkage if treatment is to be successful beyond palliative applications. The influence of an angular component in any such trauma must be emphasized; force vectors then attack the craniomandibular/temporo-mandibular/cervical complex where it is most vulnerable, at the most restrictive parameters of functional mobility. Thus, the angular component brings an exponential increase in tissue damage potential to all of the craniomandibular/temporomandibular/cervical complex. Finally, if we are to treat the occlusion of these victims intelligently, we must understand the effects of this trauma on the 'whole body', and not just focus singularly on the restoration or the malocclusion or the TMJoint problem. There are lessons to be learned from this acute trauma which provide valuable insights into the diagnosis of chronic pain patients. If these victims do not come to us for help immediately following the acute trauma episode, but arrive in our offices months or years later, our clinical examinations must include head and neck mobility or functional restrictions, observations of the patients' gait, other residual postural deficits, et al. Failure to implement these observations in our clinical examination will ultimately compromise our treatment success. As a final note, an axiom to bear in mind when studying the function of the musculoskeletal system is that when muscle is put into an adversarial relationship with bone, muscle always wins! Example #1: Compare a lateral cervical radiograph taken on the day of a rearend collision to one completed five months later; if tissue damage is unresolved, there will inevitably be a loss of lordotic curve of the cervical spine with a kyphosis at C 4, C-5!; Example #2: If a patient has developed a parafunctional habit like bruxism using pathological forces over a long period of time, the clinician will inevitably observe antigonial notching on the mandible at the masseter insertion! Although at first glance integrating these precepts into our examination protocols may appear to complicate matters, it actually makes our tasks easier and allows us to provide more comprehensive services for our patients. Those of us who are "functionally aware", will always be three steps ahead of the field in functional orthopedics or functional orthodontics. Let me conclude with a premise I learned many years ago from a very learned Osteopath, Dr. John Harakal, of Fort Worth, Texas: "If, as a clinician, you are able to put the body into a position to heal, it will heal itself." PMID- 9863427 TI - TMD warning sign: cephalometrics. AB - The significance of predisposing factors to TMD is quite evident when considering the prevalence of orthodontic relapse and with the occurrence of TMD related symptoms observed in previously treated orthodontic patients. In using the Modified Sassouni Plus Cephalometric Analysis, the clinician now possesses an invaluable tool that is readily available without adding to the examination expense. The invaluable information gained from cephalometric evaluation should not, however, be used to replace other diagnostic TMJ radiographs. If a joint problem is suspected, one should follow the normal protocol for the examination of the temporomandibular joints. PMID- 9863428 TI - Finishing touches for beautiful smiles: orthodontist and general dentist coordinate efforts to achieve optimum esthetics. Part II. AB - It is not uncommon for teeth to still not be attractive, even though they are straight following treatment. Several examples have been given of what can be done to improve the attractiveness of teeth when this occurs. It is the responsibility of the dental team involved in the management of a persons total dental care to advise patients of what dental services are available to them. People should have the opportunity to evaluate what dental services are needed to accomplish a pleasing result that is satisfying to them and to the dental team involved. PMID- 9863429 TI - Multidisciplinary management of the cervicocranial and temporomandibular disorder: appreciation of the whole Cube--clinical case. Part II. PMID- 9863430 TI - Reporting requirements for adult and child abuse. PMID- 9863431 TI - Providing patient-friendly financial options. AB - Having flexible payment options ensures that patients will have all of the motivation and information that they need to accept care in your office. An HCCC simply offers you the chance to make more expensive treatment available to those who need or want it. The small cost of using a HCCC program more than pays for itself--in fact, it costs less than one-third of what you would spend if you were to bill these patients instead. Having consistent financial policies and flexible payment options can dramatically increase your office's productivity and profitability, while expressing your commitment to customer service and high quality dentistry. PMID- 9863432 TI - Reflections on the Michigan splint and other intraocclusal devices. AB - It seems obvious in retrospect that the treatment of disorders by interocclusal devices followed two paths: stabilization splints and functional orthopedic appliances. The dividing line between them is not always clear. Both have some function related to the position of the mandible. They may not differ significantly in their control of occlusal stability (e.g., telescoping devices anchored to stabilization splints). The stabilization splint, as well as other conservative measures, will play an increasing role in accepted therapy for TMD. The use of anterior repositioning devices for TMD, including MPD syndrome, will decrease. Research may provide answers that allow them to be used more specifically and predictably. Perhaps there will be but little change in their use where there is an association of TMD and Class II malocclusion. There will be an increase in the use of interocclusal devices for the treatment of snoring and obstructive apnea. Some additional directions seem to have emerged in the late 1980s and early 1990s: In the absence of pain and significant debilitation, treatment for TMD, if any, is to be reversible. Prevention or aggravation of TMD should be practiced to the extent possible during dental procedures. One long term, well-designed, prospective study indicated that the incidence and severity of TMD could be reduced by appropriate occlusal adjustment. There is a small, but nevertheless important minority of patients with TMD who progress to persistent pain and/or dysfunction. Initial management of the vast majority of patents with TMD should be use of noninvasive reversible therapies. Surgery is indicated in only a relatively small percentage of cases of TMD. Research on interocclusal devices should not terminate simply because they are in part dental devices (i.e., biomechanical forms of treatment). The diagnosis and treatment of TMD has been called a dilemma, especially for those patients with chronic pain for whom no treatment has been effective. However, it would be ill-advised to abandon what treatment is already known to be effective by allowing those few but psychosocially important patients with chronic pain to determine what should be done for the vast majority of patients with TMD: reversible forms of treatment, including physiotherapy, pharmacologicals, and the stabilization occlusal bite plane splint. PMID- 9863433 TI - Bragdon vs. Abbott. PMID- 9863435 TI - Why this year's Supreme Court elections are so important. PMID- 9863434 TI - Hand asepsis--it's important. PMID- 9863436 TI - Renew your staff's focus with an off-site retreat. PMID- 9863437 TI - The American Dental Association/Michigan Dental Association standards of ethics and Code of Professional Conduct. PMID- 9863438 TI - Here's a staff incentive bonus program that really works! PMID- 9863439 TI - Treatment outcome for a sample of patients with Class II division 1 malocclusion treated at a regional hospital orthodontic department. AB - This retrospective study assessed the outcome of orthodontic treatment of 264 patients with Class II division 1 malocclusion (overjet greater than 6 mm). The sample comprised patients who had completed their fixed appliance orthodontic treatment at a regional hospital orthodontic unit in the Republic of Ireland. The PAR Index (Peer Assessment Rating) was used to evaluate treatment outcome using before and after treatment study casts. The results revealed that treatment for this particular type of malocclusion was highly effective with a very few patients failing to benefit from their orthodontic treatment. PMID- 9863441 TI - The death of a professional school: does anyone really care? PMID- 9863440 TI - Provision of sugar-free medicines for young children: the views of a sample of parents in Northern Ireland. AB - Sugar in liquid medicines is a factor in dental caries for many children. There has been an increase in the availability of sugar-free medicines in recent years, but they need to be prescribed specifically, because most generic liquid medicines still contain sugar. The object of the present study was to examine parental and child factors which may be linked to the provision of sugar-free alternatives by their medical practitioner. A random sample of 291 parents of children aged four years was obtained. A questionnaire was used to ascertain the extent of their knowledge concerning liquid medicines and their effect on dental health and the frequency with which such medicines are requested by them was assessed. The levels of provision of sugar-free medication and the frequency of used common types of medication were recorded and long term users identified. There was a total of 212 children in the study representing an inclusion rate of 73 per cent of the original sample. Just more than 17 per cent were always provided with a sugar-free liquid medicine, 58.3 per cent sometimes received this form of medication and 24.6 per cent never received it. The strongest individual predictor of a high frequency of provision of the medication in sugar-free form was the frequency of parents requesting this type of medication from their medical doctor and the next strongest predictor was the frequency of use of antibiotics. Parental awareness or knowledge of the importance of the issue for dental health did not affect the outcome in the present study. It is concluded that there is a need for specific campaigns to promote the use of sugar-free medications and further increases in their availability should be sought from manufacturers. PMID- 9863443 TI - Effects of sealers and liners on marginal leakage of amalgam and gallium alloy restorations. AB - In an in vitro study, the use of sealers and liners (Fuji varnish, Vitrabond, Vitremer, Paama 2, All-Bond 2, or Resinomer) significantly reduced the amount of marginal leakage around amalgam (Permite C or Lojic Plus) and gallium (Galloy) alloy restorations. This reduction in marginal leakage was produced by all sealers and liners tested, and there were no statistically significant differences between these materials. Unlined restorations of Permite C had significantly less marginal leakage than Galloy or Lojic Plus. Unlined Lojic Plus restorations had the greatest amount of marginal leakage. The experimental method used in the present study proved to be suitable for quantitative comparison of marginal leakage of different dental materials. PMID- 9863442 TI - Fluoride-releasing dental restorative materials. AB - In the 1940s, dentists observed that secondary caries was rarely associated with silicate cement restorations. While the relatively stable dimensional properties of those restorations were undoubtedly a factor in this fortunate circumstance, the fact that fluoride was inherent in the composition of the material received the greater credit. By the mid-1980s, a wide variety of fluoride-releasing dental restorative materials were available to dentists and dental consumers, and the cariostatic effect of fluoride ions on enamel caries had been demonstrated in many studies. This paper reviews much of the fluoride-related research conducted on fluoride-releasing amalgam, glass-ionomer cements, composites, primers, sealants, liners, acrylic resins, and orthodontic bracket bonding materials. The need for standardization of test methods is addressed, as is the need for more controlled clinical trials and additional research. PMID- 9863444 TI - Selective caries removal with air abrasion. AB - The purpose of this study was to evaluate selective caries removal using an air abrasive technique. Alumina powders, glass beads, crushed glass powders, and crushed powders of polycarbonate resin were applied to intact human enamel, dentin, and artificially demineralized dentin (caries-model dentin). Furthermore, the effect of the particle size of abrasives and air pressures on the abraded depths was examined. When alumina powders and glass beads were used, the abraded depths of enamel, dentin, and caries model increased as the particle size and air pressure increased. Alumina powders and crushed glass powders abraded intact enamel and dentin more than the caries-model dentin, whereas glass beads abraded the caries-model dentin more than the intact enamel and dentin. Only crushed powders of polycarbonate resin abraded the caries-model dentin without reducing intact enamel and dentin. With hard particles, such as alumina powders, glass beads, or crushed glass powders, selective caries removal by the air-abrasive technique appeared to be difficult to achieve, even if the particle size and the air pressure were changed. Crushed powders of polycarbonate resin that reduced only the caries-model dentin are harder than caries-model dentin, but softer than intact enamel and dentin. This study elucidated the possibility of selective carious dentin removal with the air-abrasive technique. PMID- 9863445 TI - Evaluation of acidic primers in microleakage of Class 5 composite resin restorations. AB - The purpose of this in vitro study was to evaluate the marginal seal of two adhesive systems on class 5 composite resin restorations. Two adhesive systems that use acidic primers were used in this study: Clearfil Liner Bond 2 and Denthesive II. Scotchbond Multi-Purpose was used as a control. Class 5 cavity preparations were made at the CEJ (half of the preparation was in enamel and half in dentin/cementum) of 60 extracted human molars. Both systems were used with and without etching. The control group was used with etching only. Composite resin restorations were placed, light cured for 40 seconds, and polished. All specimens were thermocycled 500 times. The specimens were immersed in erythrosin B, tetra iodo-fluorescent 2% solution, then sectioned to obtain 600 mm-thick sections. These were examined under X13.2 magnification, and the degree of dye penetration was measured in microns. A chi-square test demonstrated no significant differences in microleakage among groups for either dentin (P = 0.54) or enamel (P = 0.35). PMID- 9863446 TI - Effect of etchant, etching period, and silane priming on bond strength to porcelain of composite resin. AB - The purpose of this study was to evaluate the effect of etching and silane priming on bond strength to a feldspathic porcelain (VMK 68) of a composite resin (Clearfil APX). Two hydrofluoric acid etchants (2.5% and 5%) and seven different etching times (0, 30, 60, 90, 120, 150, and 180 seconds) were used to etch the porcelain specimens respectively. A self-curing bonding agent containing a silane coupler (Clearfil Porcelain Bond) was used on both etched and unetched porcelain surfaces. Etched relief patterns were observed by means of a scanning electron microscope, and the bond strengths between the photocured composite resin and the porcelain were determined. Scanning electron micrographs revealed complicated etching patterns with increased etching time periods. Shear testing results showed that the bond strength to the unetched porcelain of the composite resin was very low, and that etching periods for more than 30 seconds effectively enhanced the bond strength. Of the two etching agents applied to the unsilanated porcelain, the buffered 2.5% etchant produced higher bond strengths than the 5% etchant for all etching time periods except for 180 seconds. Silane priming was effective and critical for improving bond strength to the porcelain. Application of the silane bonding agent to the porcelain after hydrofluoric acid etching appeared to be suitable for achieving consistent bonding between the composite resin and the porcelain. PMID- 9863447 TI - Influence of air-abrasion treatment on the interfacial bond between composite and dentin. AB - The purpose of this in vitro study was to investigate the effect of air abrasion on the interfacial bond between composite and dentin. Dentin surfaces surrounded by enamel were produced by removing the incisal part of the crown from extracted human anterior teeth. The dentinal area of the specimen surfaces was conditioned either by air-abrasion treatment (27 microns aluminum oxide particles, 120 psi), application of dentin adhesives, and combination of air abrasion and dentin adhesives, or left nontreated (controls). The enamel was conditioned by acid etching, by air abrasion, or by air abrasion and subsequent acid etching. A 2 mm layer of six different composite resins (Brilliant, Charisma, Helioprogress, Herculite, Pekafill, Z-100) was applied on the specimen surfaces and light cured. The composite dentinal interface was evaluated in cross-cut sections of 324 specimens by SEM analysis, and the mean gap width (MGW) between composite and dentin was calculated. SEM analysis revealed gap formation ranging from 1.1-3.6 microns in control specimens. Gap width was reduced significantly due to air abrasion treatment of the dentinal surface (MGW: 0.3-1.4 microns) or by application of dentin adhesives (MGW: 0.06-0.6 micron). Combined air-abrasion treatment and application of dentin adhesives resulted in a gap-free adaptation between composite and dentin in the majority of specimens. However, thermocycling of the specimens (5 degrees C/55 degrees C; 1000 cycles) caused a significant increase in gap formation (MGW: 0.03-2.9 microns) at the composite-dentin interface. It is concluded that air-abrasion treatment and subsequent application of a dentin adhesive is an effective procedure to resist the contraction stress at the dentinal surface during polymerization of the composite resin and to improve the internal bond of composite resin restorations. PMID- 9863448 TI - Color attributes and accuracy of Vita-based manufacturers' shade guides. AB - The shades of several tooth-colored restoratives are now keyed to the Vita Lumin shade guide. The purpose of this study was to determine whether manufacturers' shade guides had color attributes that were similar to the Vita Lumin guide to which they are supposedly keyed. The overall color accuracy between the different guides was also compared via colorimetry. Results showed that none of the manufacturers' shade tabs evaluated had all L*, a*, and b* values that were identical to their respective Vita shade tabs. For the shades evaluated, manufacturers' tabs were generally darker than their corresponding Vita shade tabs. When shade guides were compared, the color accuracy of Z100 and Fuji II LC shade guides was significantly better than that of Durafill VS and TPH. Dyract's shade guide was the most accurate and was significantly better than the shade guides of Fuji II LC, Durafill VS, and TPH. The accuracy of manufacturers' shade guides was not consistently product dependent but was shade dependent. PMID- 9863449 TI - Change in size of replaced amalgam restorations: a methodological study. AB - The purpose of this study was to evaluate the pre- and postoperative size of amalgam restorations that were scheduled for replacement. A video imaging system in combination with specially designed devices to align the stone casts prepared from impressions of the teeth gave reproducible results for both an in vitro and in vivo series. The in vivo series showed that removal of the occlusal part of amalgam restorations could be done without significantly increasing the size of the cavity in the tooth, provided the restorations did not have marginal discrepancies. If marginal discrepancies or recurrent caries were present, the replacement restorations showed a statistically significant increase in size compared with the original restorations. PMID- 9863450 TI - Periostat: low-dose doxycycline. A commentary. PMID- 9863451 TI - Gene therapy. PMID- 9863452 TI - A multidisciplinary approach to the treatment of oligodontia: case report. AB - The treatment of young people with implants requires advanced planning and coordination of many different specialities within dentistry. Timing and sequence of therapy will often decide the success or failure of treatment. Congenital absence of many teeth (oligodontia) associated with or without syndromes poses not only functional but also psychosocial problems for young people. A case is presented in which orthodontist, restorative dentist, and periodontist evaluated and performed the necessary therapy for a young person who at the age of 9 was diagnosed with oligodontia/l. Nine implants were placed: 13 years, 8 months in the mandible and 15 years, 6 months in the maxilla. All implants were restored as single teeth. The patient was followed until age 20. PMID- 9863453 TI - Restoration of localized severely atrophic maxillary ridge: case report. AB - A procedure is described whereby a patient with a highly localized atrophic maxillary ridge has had the deformity corrected by utilizing a combination of a block of corticocancellous bone and a barrier membrane. PMID- 9863454 TI - Preservation of the alveolar ridge at implant sites. AB - This case report describes an alternative surgical approach which can avoid the need for secondary regenerative techniques for ridge augmentation when planning treatment for dental implants. Our patient had a hopeless maxillary left central incisor, which was subsequently extracted in preparation for implant restoration. The site had a significant defect, which was treated at the time of extraction with a combination of a xenograft and a cortical chin graft. The goal was to preserve the residual ridge contour to avoid a compromise in implant fixture placement and maximize function and esthetics. PMID- 9863455 TI - Jury duty. 1893. PMID- 9863456 TI - A common denominator to veneers and all-ceramic crown restorations. PMID- 9863457 TI - Treatment considerations for posterior laboratory-fabricated composite resin restorations. AB - A variety of techniques are currently available to restore dentition in the posterior region, and the selection of the proper modality is dependent upon evaluation and satisfaction of numerous criteria. Direct materials provide limited strength in the posterior segment, and full-coverage indirect techniques (e.g., crown restorations) often require the destruction of sound tooth structure. When proper case selection, preparation design, and clinical protocols are utilized, a laboratory-processed composite resin (belleGlass, Kerr/Sybron, Orange, CA) can provide an optimal means by which to successfully restore posterior dentition without compromising the existing tooth structure. PMID- 9863458 TI - Interdisciplinary posterior restoration with a single-tooth onlay. PMID- 9863459 TI - The role of research in clinical practice. PMID- 9863460 TI - Subattachment inflammatory root resorption: treatment strategies. AB - Attempts have been made to classify root resorption according to its etiology in order to develop more efficient strategies for its treatment. This condition has been identified as resulting from damage to the protective attachment layer (predentin or precementum) of the root combined with inflammation adjacent to the root surface. This article demonstrates a series of treatment strategies focused on the reversal of each manifestation of subattachment root resorption, specifically emphasizing defects that extend apically and localized defects. PMID- 9863461 TI - Advances in digital radiology--corded and cordless systems. PMID- 9863462 TI - Implant placement without flaps: a single-stage surgical protocol--Part 1. AB - Osseointegrated implants have been demonstrated to be predictable, efficacious means of restoring aesthetics and long-term function to edentulous patients. Although the surgical and placement protocols for the use of implant therapy have continued to evolve, specific criteria must be addressed when utilizing this modality. The objective of this article is to describe the requisites for proper implant placement and demonstrate the use of a novel surgical protocol to insert implant fixtures in selected edentulous sites with a mucosal tissue punch rather than a conventional flap elevation procedure. PMID- 9863463 TI - Motor vehicle accidents: developing an accurate differential diagnosis. PMID- 9863464 TI - Anterior fixed prosthetic restorations and the bleached dentition: laboratory techniques. AB - The process of matching fixed prosthetic restorations to bleached dentition has often created a dilemma, as clinicians are not able to accurately communicate shade selection to the dental laboratory. By understanding the effect of vital bleaching on the dentition, it is possible to fabricate porcelain restorations that match the bleached natural teeth. This article presents a laboratory technique for utilizing a sequence of porcelain powders and stains to develop the internal anatomy (e.g., halos, mamelons, luminosity) of restorations that appear indistinguishable from the bleached dentition. PMID- 9863465 TI - Contemporary developments in the utilization of metal-ceramic restorations: a review. AB - Numerous porcelain systems and techniques are currently available for utilization in full-coverage anterior crown restorations. Significant improvements have been made in the placement of margins and the reduction of the metal framework in order to enhance the optical effects (e.g., illumination, warmth, and vitality) of these porcelain systems. This article presents several recent developments in the fabrication of metal-ceramic crown restorations, and demonstrates the use of these principles in the aesthetic oral rehabilitation. PMID- 9863466 TI - Ileal pouch-anal anastomosis for Crohn's disease. PMID- 9863467 TI - Sphincter of Oddi dysfunction and acute pancreatitis. PMID- 9863468 TI - Making light work of early gastric cancer. PMID- 9863469 TI - Differentiating ulcerative colitis from Crohn's disease: false dawn for CD44. PMID- 9863470 TI - The kappa opioid receptor is associated with the perception of visceral pain. AB - mu-, delta- and kappa-opioid receptors are widely expressed in the central nervous system where they mediate the strong analgesic and mood-altering actions of opioids, and modulate numerous endogenous functions. To investigate the contribution of the kappa-opioid receptor (KOR) to opioid function in vivo, we have generated KOR-deficient mice by gene targeting. We show that absence of KOR does not modify expression of the other components of the opioid system, and behavioural tests indicate that spontaneous activity is not altered in mutant mice. The analysis of responses to various nociceptive stimuli suggests that the KOR gene product is implicated in the perception of visceral chemical pain. We further demonstrate that KOR is critical to mediate the hypolocomotor, analgesic and aversive actions of the prototypic kappa-agonist U-50,488H. Finally, our results indicate that this receptor does not contribute to morphine analgesia and reward, but participates in the expression of morphine abstinence. Together, our data demonstrate that the KOR-encoded receptor plays a modulatory role in specific aspects of opioid function. PMID- 9863471 TI - Annual colonoscopy, chest radiography, and computed tomography of the liver did not prolong survival in patients with colorectal cancer. PMID- 9863472 TI - Naproxen sodium did not lead to substantially more upper gastrointestinal tract bleeding than ibuprofen during short term use as an analgesic. PMID- 9863473 TI - Exposure to mesalamine during pregnancy increased preterm deliveries (but not birth defects) and decreased birth weight. PMID- 9863474 TI - Helicobacter pylori specific nested PCR assay for the detection of 23S rRNA mutation associated with clarithromycin resistance. AB - BACKGROUND: Clarithromycin is one of the most important antibiotics for Helicobacter pylori eradication. However, 5-10% of strains are reported to be resistant. It has been shown that one point mutation in the 23S rRNA gene is associated with resistance to clarithromycin. AIMS: To establish a polymerase chain reaction (PCR) system which amplifies a segment of the 23S rRNA gene containing the mutation points with primers specific for H pylori, so that H pylori infection and the mutation associated with clarithromycin resistance can be examined simultaneously. METHODS: To detect H pylori infection and the mutation simultaneously, primers specific for the H pylori 23S rRNA gene were designed based on sequence conservation among H pylori strains and sequence specificity as compared with other bacteria. DNA from 57 cultured strains and from 39 gastric juice samples was amplified in the seminested 23S rRNA PCR. Clinical applicability was evaluated in 85 patients. RESULTS: DNA samples from 57 cultured strains were all amplified. The novel assay and the urease A PCR agreed in 37/39 gastric juice samples with no false positives. The assay did not amplify the DNA of bacteria other than H pylori. Eight of 85 samples had the mutation before treatment. In clarithromycin based treatment, eradication was achieved in 2/5 (40%) with the mutation and 29/34 (85%) without the mutation. CONCLUSION: The assay using gastric juice is quick (within 12 hours) and non-invasive (endoscopy not required), enabling rapid initiation of appropriate antibiotic treatment. PMID- 9863475 TI - The relation between gastric vitamin C concentrations, mucosal histology, and CagA seropositivity in the human stomach. AB - BACKGROUND: Vitamin C may be protective against gastric cancer though infection with Helicobacter pylori is associated with a reduction in intragastric concentrations of vitamin C. AIMS: To examine the effects of H pylori infection, gastric juice pH, the severity and extent of gastric inflammation, and CagA antibody status of the individual on gastric juice and mucosal vitamin C concentrations. PATIENTS: One hundred and fifteen patients undergoing routine gastroscopy for investigation of dyspepsia. METHODS: High performance liquid chromatography was used to determine vitamin C concentrations. CagA antibody was detected by western blot analysis. RESULTS: Gastric juice ascorbic acid concentration was significantly lower in patients infected with H pylori compared with those uninfected (19.3 mumol/l (interquartile range (IQR) 10.7-44.5) versus 66.9 mumol/l (IQR 24.4-94.2), p = 0.003). The reduction in gastric juice ascorbic acid concentration was inversely related to the severity of gastritis (p = 0.01). CagA positive patients had significantly lower gastric juice ascorbic acid concentrations than CagA negative ones (14.8 mumol/1 (IQR 7.9-52.2) versus 39 mumol/l (IQR 19.9-142.2), p = 0.05). Decreased gastric juice dehydroascorbic acid concentrations were observed in patients with gastric atrophy and intestinal metaplasia. Mucosal ascorbic acid concentrations were also significantly lower in infected patients than uninfected patients (p = 0.04). CONCLUSIONS: The reduction in gastric vitamin C concentrations is related to gastric juice pH, the severity and extent of gastritis, the presence of H pylori, and the CagA antibody status of the individual. These findings may have implications in H pylori associated carcinogenesis. PMID- 9863476 TI - Opposing time trends of peptic ulcer and reflux disease. AB - BACKGROUND: Gastritis associated hypochlorhydria may be protective against gastro oesophageal reflux disease. It was hypothesised that the historic decline in Helicobacter pylori infection resulted in a decline in peptic ulcer and a concomitant risk in reflux disease. AIMS: To study the time trends of peptic ulcer and reflux disease. METHODS: Hospitalisation rates were analysed using the computerised database of the US Department of Veterans Affairs from 1970 until 1995. Death rates were calculated from the Vital Statistics of the United States. RESULTS: From 1970 to 1995, hospitalisation rates for gastric and duodenal ulcer, as well as gastric cancer fell, while the hospitalisation rates for gastro oesophageal reflux disease and those for oesophageal adenocarcinoma rose significantly. Similar time trends were observed with respect to the death rates. There were notable ethnic differences. White subjects incurred higher rates of reflux disease and oesophageal adenocarcinoma and lower rates of gastric ulcer or cancer compared with non-whites. CONCLUSIONS: The ethnic variations and the opposing time trends of gastroduodenal versus oesophageal disease are consistent with the hypothesis that the declining infection rates of H pylori in the general population have led to a rise in the occurrence of gastro-oesophageal reflux disease and associated oesophageal adenocarcinoma. PMID- 9863478 TI - Early gastric stump cancer following distal gastrectomy. AB - BACKGROUND: Gastric stump cancer (GSC) is usually diagnosed at an advanced stage, and consequently the prognosis is poor. AIMS: To investigate the clinicopathological characteristics of GSC at an early stage to assist in its identification, and thereby improve its prognosis. METHODS: Forty three patients with resected early GSC were compared with 156 patients with resected primary early cancer in the upper third of the stomach. RESULTS: Sixty five per cent (28/43) of the early GSC patients showed the elevated type endoscopically, although the frequency of the depressed type in GSC has tended to increase in the past five years. This occurred in less than 26% (40/156) of the primary early cancers. Half of the early GSCs were located on the lesser curvature (47%), and revealed differentiated adenocarcinoma (81%) histologically. The male:female ratio of early GSC cases was about 6:1, which was much higher than that in patients with primary early cancer. The five year survival rates of patients with early GSCs and early primary cancers were 84% and 95%, respectively. GSC had a favourable prognosis, if it was detected at an early stage. CONCLUSION: To detect early GSC, our results suggest that special attention should be given to elevated as well as depressed lesions on the lesser curvature of the stomach, particularly in men, during endoscopic examinations. PMID- 9863477 TI - Pentagastrin gastroprotection against acid is related to H2 receptor activation but not acid secretion. AB - BACKGROUND: Pentagastrin enhances gastric mucosal defense mechanisms against acid and protects the gastric mucosa from experimental injury. AIMS: To investigate whether this gastroprotection is mediated by histamine receptors or occurs as a secondary effect of acid secretion stimulation. METHODS: The effects of omeprazole (100 mumol/kg), ranitidine (20 mg/kg), and pyrilamine (10 mg/kg) on pentagastrin (80 micrograms/kg/h) induced gastroprotection against acidified aspirin injury were examined in a luminal pH controlled model. The effects of these compounds on pentagastrin enhanced gastroprotective mechanisms were investigated using intravital microscopy, in which intracellular pH of gastric surface cells (pH1), mucus gel thickness, gastric mucosal blood flow, and acid output were measured simultaneously. RESULTS: Pentagastrin protected rat gastric mucosa from acidified aspirin injury. This gastroprotection was abolished by ranitidine, but not omeprazole or pyrilamine. Pentagastrin induced a hyperaemic response to luminal acid challenge, increased mucus gel thickness, and elevated pHi during acid challenge. Ranitidine reversed these enhanced defence mechanisms, whereas omeprazole and pyrilamine preserved these effects. CONCLUSIONS: These data indicate that pentagastrin associated gastroprotection and enhanced defence mechanisms against acid result mainly from activation of histamine H2 receptors, and not as an effect of the stimulation of acid secretion. PMID- 9863479 TI - Photodynamic ablation of early cancers of the stomach by means of mTHPC and laser irradiation: preliminary clinical experience. AB - BACKGROUND: Radical surgical treatment has been the first choice for early gastric cancer, but high resolution endosonography allows in situ diagnosis with a high sensitivity and specificity and in consequence the option of local endoscopic treatment. AIMS: To evaluate photodynamic therapy (PDT) using meso tetrahydroxyphenyl-chlorin (mTHPC) as the photosensitiser in superficial gastric cancer. METHODS: Twenty two patients with superficial early gastric cancer received mTHPC (0.075 mg/kg intravenously) and were treated 96 hours later with red light at 652 nm (20 J/cm2). Tumour response was assessed by endoscopy and extensive biopsy. RESULTS: Endoscopies performed two to three days after PDT showed a haemorrhagic fibrinoid necrosis of the mucosal layer. Complete remission was achieved in 16/22 (73%) patients: 13/16 (80%) with intestinal type cancer and 3/6 (50%) with a diffuse Lauren's carcinoma. The mean follow up period was 12 months and 20 months, respectively. An average of 1.8 treatment sessions was required. Severe side effects were not observed. Seven patients had mild to moderate skin photo-sensivity reactions; 12 had local pain after PDT for 1-10 days. CONCLUSION: PDT using mTHPC as the photosensitiser represents a safe and efficient method for topical treatment of early gastric cancer, especially of Lauren's intestinal carcinoma. If the preliminary results can be verified in larger patient series and during long term follow up, local treatment of early cancers of the stomach by PDT could be considered as a therapeutic option for selected patients. PMID- 9863480 TI - HIV enteropathy: comparative morphometry of the jejunal mucosa of HIV infected patients resident in the United Kingdom and Uganda. AB - AIMS: To compare jejunal mucosal morphometry in HIV infected patients resident in London and Uganda. PATIENTS: Twenty HIV positive patients from London and 16 from Uganda were studied, and compared with HIV negative control subjects from both sites. METHODS: Stools and biopsy specimens were examined for enteropathogens. Surface area to volume (S:V) ratio was estimated morphometrically, mean crypt length of jejunal biopsy specimens was measured, and HIV infected cells detected immunohistochemically were quantified. RESULTS: Enteric pathogens were detected in none of the London patients, and in three Ugandan patients. S:V ratio was lower, and mean crypt length higher, in the specimens of London patients than in normal subjects, but there was no difference in S:V ratio or mean crypt length between Ugandan patients and controls. A negative correlation was present between S:V ratio and mean crypt length in all biopsy specimens analysed. HIV infected cells were detected only in lamina propria. CONCLUSION: Infection of cells in the lamina propria of the jejunum with HIV stimulates crypt cell proliferation, and a fall in villous surface area. The mucosal response to HIV is masked by other pathogens in the African environment. PMID- 9863481 TI - Correlation of desensitisation of platelet activating factor (PAF) receptors with intensity of inflammation and intestinal PAF content during experimental ileitis in guinea pig. AB - AIM: To determine the kinetics of platelet activating factor (PAF) and prostaglandin E2 (PGE2) receptor desensitisation during intestinal inflammation induced by trinitrobenzenesulphonic acid (TNB) instillation and to study the relation between receptor regulation, inflammatory lesions, and PAF content of the gut wall. METHODS: Receptor desensitisation was assessed on isolated smooth muscle cells from the circular layer. PAF content of the intestinal wall was determined by thin layer chromatography and radioimmunoassay. RESULTS: After an acute inflammatory phase on day 1, subacute changes appeared in TNB instilled ileum, with a maximal intensity on day 6. In control animals, PAF 10 nM and PGE2 10 nM provoked a maximal contraction in the range of 24% of cell shortening. On days 1 and 3 after intestinal instillation of TNB, PAF induced contraction was not altered whereas the effect of PGE2 was progressively desensitised (2 logM rightward shift of its concentration-response curve: Cmax = 1 microM; p < 0.01). Between days 4 and 6, the concentration-response curve of PGE2 shifted by only 1 logM (p < 0.05) whereas the curve of PAF induced contraction shifted by 2 logM (Cmax = 1 microM; p < 0.01). The PAF content of the ileal wall was maximal between days 3 and 5 (300 ng/mg tissue). On days 10 and 15, PAF and PGE2 induced contractions were similar to those observed on day 1, and PAF content returned to basal. CONCLUSION: Inflammation induced by TNB instillation triggers PAF and PGE2 receptor desensitisation; this is dependent on the duration of inflammation and correlates with PAF content in the ileum. This receptor desensitisation may play a protective role by preventing overstimulation of intestinal smooth muscle cells. PMID- 9863482 TI - Kallikrein-kininogen system activation and bradykinin (B2) receptors in indomethacin induced enterocolitis in genetically susceptible Lewis rats. AB - BACKGROUND: The plasma kallikrein-kinin (K-K) system is activated in acute and chronic relapsing intestinal inflammation induced in Lewis rats by intramural injection of exogenous bacterial components. AIMS: To determine whether this effect is model specific, K-K system activation was investigated in a modified indomethacin induced enterocolitis model, as well as bradykinin 2 (B2) receptor distribution in the normal and acutely inflamed intestine. METHODS: Lewis rats injected with daily sublethal doses of indomethacin for two days developed acute (two days) and chronic (14 days) intestinal inflammation. Plasma prekallikrein (amidolytic), high molecular weight kininogen (HK, coagulant) and cleavage of HK (western blot) were assayed to detect K-K activation. RESULTS: Liver and spleen weights were significantly higher, and body weights and haematocrit values were significantly lower in the indomethacin group than in the control group. During both acute and chronic phases, rats displayed K-K system activation manifested by a significant decrease in plasma prekallikrein and HK functional levels, and by HK cleavage. Plasma T kininogen (a major acute phase protein) was significantly elevated. B2 receptors were identified in both normal and inflammatory intestine with more prominent specific immunohistochemical staining in the acutely inflamed tissue. CONCLUSIONS: K-K system activation occurs in association with both acute and chronic phases of intestinal injury, regardless of the triggering agent, suggesting that activation of this system is integrally involved in intestinal inflammation in genetically susceptible hosts. Localisation of B2 receptors across intestinal layers provides a structural basis for the kinin function in the intestine. PMID- 9863483 TI - Poor diagnostic value of colonic CD44v6 expression and serum concentrations of its soluble form in the differentiation of ulcerative colitis from Crohn's disease. AB - BACKGROUND: Increased expression of CD44v6 on colonic crypt epithelial cells in ulcerative colitis has been suggested as a diagnostic tool to distinguish ulcerative colitis from colonic Crohn's disease. AIMS: To investigate colonic CD44v6 expression and serum concentrations of soluble CD44v6 (sCD44v6) in patients with ulcerative colitis and Crohn's disease. METHODS: Colonic biopsy samples were obtained from 16 patients with ulcerative colitis, 13 with ileocolonic Crohn's disease, and 10 undergoing polypectomy. Serum samples were obtained from 15 patients with active ulcerative colitis, 20 with active Crohn's disease, and 20 healthy donors. Colonic CD44v6 expression was evaluated immunohistochemically by monoclonal antibody 2F10 and the higher affinity monoclonal antibody VFF18. Serum sCD44v6 concentrations were measured by ELISA. RESULTS: 2F10 stained colonic epithelium of inflamed ulcerative colitis and Crohn's disease samples in 80% and 40% of cases, respectively, and VFF18 in 95% and 87%, respectively. Both monoclonal antibodies displayed a sensitivity and specificity of 60% and 87% to differentiate ulcerative colitis from colonic Crohn's disease. Serum concentrations of sCD44v6 were lower in patients with ulcerative colitis (median 153 ng/ml; interquartile range (IQR) 122-211) compared with Crohn's disease (219; IQR 180-243) and healthy donors (221; IQR 197-241 (p = 0.002)). Its sensitivity and specificity to discriminate ulcerative colitis from Crohn's disease was 75% and 71%, respectively. CONCLUSION: Colonic CD44v6 and serum sCD44v6 concentrations do not facilitate reliable differential diagnosis between ulcerative colitis and Crohn's disease. PMID- 9863484 TI - Loss of Bcl-2 expression correlates with tumour recurrence in colorectal cancer. AB - AIMS: To investigate the association between immunohistochemical expression of Bcl-2 and p53 in colorectal cancer and tumour recurrence following surgery. METHODS: Sixty six cases of Dukes' B colorectal carcinoma were studied. All tumours were moderately differentiated and were shown to be histologically clear of the resection margins. Immunohistochemistry was performed on formalin fixed paraffin wax embedded tissue using monoclonal antibodies for p53 and Bcl-2. The Bcl-2 staining was assessed separately for relative intensity of staining and percentage of positive tumour cells and given a final score which combined the two factors. The p53 staining was assessed on number of positive tumour cells only. The patterns of immunostaining of those cases in which there had been tumour recurrence were compared with those cases in which there was no tumour recurrence (controls). RESULTS: A statistically significant inverse association was found between Bcl-2 score and tumour recurrence (median Bcl-2 score of 6 (interquartile range (IQR) 2-9) in patients with recurrent disease; median Bcl-2 score of 8 (IQR 6-10) in those without recurrence; p = 0.03). When examined separately, both the intensity of expression and percentage of positive tumour cells were significantly associated with tumour recurrence (p = 0.04 in each case). There was no association between p53 staining and tumour recurrence. CONCLUSION: Results suggest that, when controlled for differentiation, Bcl-2 expression is a prognostic marker and may be useful as an adjunctive test in clinical decision making. PMID- 9863485 TI - Subtypes of constipation predominant irritable bowel syndrome based on rectal perception. AB - BACKGROUND: Patients who complain of constipation can be divided into those who have lost the natural call to stool, but develop abdominal discomfort after several days without a bowel movement (no urge); and those who experience a constant sensation of incomplete evacuation (urge). AIMS: To determine whether the two groups differ in symptoms, colonic transit, and perceptual responses to controlled rectal distension. METHODS: Forty four patients with constipation were evaluated with a bowel symptom questionnaire, colonic transit (radiopaque markers), and rectal balloon distension. Stool (S) and discomfort (D) thresholds to slow ramp (40 ml/min) and rapid phasic distension (870 ml/min) were determined with an electronic distension device. Fifteen healthy controls were also studied. RESULTS: All patients had Rome positive irritable bowel syndrome (IBS); 17 were no urge and 27 urge. Mean D threshold to phasic rectal distensions was 28 (3) mm Hg in no urge, 27 (3) mm Hg in urge (NS), but higher in the control group (46 (2) mm Hg; p < 0.01). Sixty seven per cent of no urge and 69% of urge were hypersensitive for D. Slow ramp distension thresholds were higher in no urge (S: 26 (3); D: 45 (4) mm Hg) compared with urge (S: 16 (2); D: 31 (3) mm Hg; p < 0.01), or with controls (S: 15 (1); D: 30 (3); p < 0.01). CONCLUSIONS: Hyposensitivity to slow rectal distension is found in patients with IBS who complain of constipation and have lost the call to stool even though their sensitivity to phasic distension is increased. PMID- 9863486 TI - Involvement of two different pathways in the motor effects of erythromycin on the gastric antrum in humans. AB - BACKGROUND: During the interdigestive state in humans, erythromycin 40 mg induces a premature activity front that starts in the stomach, while erythromycin 200 mg induces a prolonged period of enhanced antral contractile activity. AIMS: To study the involvement of a cholinergic pathway in the motor effects of erythromycin using the muscarinic antagonist atropine and the neural 5-HT1 receptor agonist sumatriptan. METHODS: In 30 healthy volunteers, fasted antroduodenojejunal motor activity was studied by stationary manometry. Placebo (n = 10), atropine (15 micrograms/kg intravenous bolus plus 15 micrograms/kg/h over 30 minutes; n = 10), or sumatriptan (6 mg subcutaneously; n = 10) was administered, followed by infusion of erythromycin 40 mg or 200 mg. RESULTS: After placebo, erythromycin 40 mg induced a premature activity front with gastric onset after 19.1 (1.7) minutes in all volunteers. After atropine, erythromycin 40 mg failed to induce a premature activity front during a 60 minute period in all volunteers (p < 0.001), while sumatriptan prevented the induction of a premature activity front during a 60 minute period in all but one volunteer (p < 0.005). The number of antral contractions and their mean amplitude in the 60 minutes after erythromycin 200 mg did not differ significantly after atropine or sumatriptan versus placebo. CONCLUSIONS: The antral motor effects of erythromycin in humans are mediated via different pathways. The induction of a premature activity front is mediated through activation of an intrinsic cholinergic pathway, while the induction of enhanced antral contractile activity may be mediated via a pathway potentially involving activation of a muscular receptor. PMID- 9863487 TI - On the protective mechanisms of nitric oxide in acute pancreatitis. AB - BACKGROUND: Ectopic protease activation, microcirculatory changes, and leucocyte activation are the main events in the pathogenesis of acute pancreatitis. Nitric oxide (NO) is known to be a key mediator in the normal and inflamed pancreas. AIMS: To investigate the targets on which NO exerts its effect in caerulein induced pancreatitis. METHODS: Acute pancreatitis was induced in rats which additionally received either the NO synthase substrate, L-arginine; the NO donor, sodium nitroprusside; or the NO synthase inhibitor, N-nitro-L-arginine methyl ester (L-NAME). At six hours, pancreatic injury (oedema, leucocyte content, ectopic trypsinogen activation) was analysed and pancreatic oxygenation and perfusion were determined. A direct influence of NO on amylase secretion and trypsinogen activation was evaluated separately in vitro. RESULTS: Both NO donors reduced the grade of inflammation. L-NAME increased the severity of inflammation, while decreasing pancreatic tissue oxygenation. Although neither amylase secretion nor intracellular trypsinogen activation in caerulein stimulated pancreatic acini was influenced by either NO donors or inhibitors, both NO donors decreased intrapancreatic trypsinogen activation peptide (TAP) and pancreatic oedema in vivo, and L-NAME increased TAP. CONCLUSIONS: NO protects against injury caused by pancreatitis in the intact animal but has no discernible effect on isolated acini. It is likely that in pancreatitis NO acts indirectly via microcirculatory changes, including inhibition of leucocyte activation and preservation of capillary perfusion. PMID- 9863488 TI - Multisystemic production of interleukin 10 limits the severity of acute pancreatitis in mice. AB - BACKGROUND: Interleukin 10 (IL-10) decreases the severity of experimental acute pancreatitis. The role of endogenous IL-10 in modulating the course of pancreatitis is currently unknown. AIMS: To examine the systemic release of IL-10 and its messenger RNA production in the pancrease, liver, and lungs and analyse the effects of IL-10 neutralisation in caerulein induced acute pancreatitis in mice. METHODS: Acute necrotising pancreatitis was induced by intraperitoneal caerulein. Serum levels of IL-10 and tumour necrosis factor (TNF), and tissue IL 10 and TNF-alpha gene expression were assessed. After injecting control antibody or after blocking the activity of endogenous IL-10 by a specific monoclonal antibody, the severity of acute pancreatitis was assessed in terms of serum enzyme release, histological changes, and systemic and tissue TNF production. RESULTS: In control conditions, serum IL-10 levels increased and correlated with the course of pancreatitis, with a maximal value eight hours after induction. Both IL-10 and TNF-alpha messengers showed a similar course, and were identified in the pancreas, liver, and lungs. Neutralisation of endogenous IL-10 significantly increased the severity of pancreatitis and associated lung injury as well as serum TNF protein levels (+75%) and pancreatic, pulmonary, and hepatic TNF messenger expression (+33%, +29%, +43%, respectively). CONCLUSIONS: In this non-lethal model, systemic release of IL-10 correlates with the course of acute pancreatitis. This anti-inflammatory response parallels the release of TNF and both cytokines are produced multisystemically. Endogenous IL-10 controls TNF alpha production and plays a protective role in the local and systemic consequences of the disease. PMID- 9863489 TI - bax, but not bcl-2, influences the prognosis of human pancreatic cancer. AB - BACKGROUND: bcl-2 and bax belong to the bcl-2-related gene family, which marks a new class of genes that influence apoptosis. The bcl-2 oncogene acts as a broad antiapoptotic factor and extends both normal and tumour cell survival. In contrast, the bax gene is a promoter of apoptosis. AIMS: To analyse the expression of bcl-2 and bax in pancreatic cancer and correlate the results with clinical parameters. PATIENTS: Pancreatic cancer tissue samples were obtained from 28 female and 32 male patients (median age 63, range 43-79 years) having surgery for pancreatic cancer. Normal pancreatic tissues obtained from 18 previously healthy organ donors served as controls. METHODS: The levels of bcl-2 and bax mRNA expression were analysed by northern blot and the exact site of mRNA transcription was determined by in situ hybridisation. The presence of the corresponding proteins was determined by immunohistochemistry. RESULTS: Northern blot analysis indicated that, in comparison with the normal pancreas, bcl-2 mRNA was overexpressed in 30% and bax mRNA in 61% of the pancreatic cancer samples. Concomitant overexpression of bcl-2 and bax was present in 26% of the cancer samples. Pancreatic adenocarcinomas exhibited 3.7-fold and 5.4-fold increases (p < 0.001) in bcl-2 and bax mRNA levels respectively. In situ hybridisation showed that both bcl-2 and bax mRNA were expressed in the cancer cells. Immunohistochemical analysis showed positive Bcl-2 and Bax immunostaining in 28 and 83% of the cancer samples respectively. In multivariate analysis (Cox regression model), bax expression was found to be a strong indicator of survival (p < 0.001). Patients whose tumours exhibited Bax immunostaining lived significantly longer (12 months) than those whose tumours were Bax negative (five months) (p < 0.039). In contrast, no relation was found between Bcl-2 and survival time. CONCLUSIONS: The data indicate that genes that are involved in the regulation of apoptosis are upregulated in human pancreatic cancer cells. Prolonged survival times in patients in whom apoptosis promoting factors are upregulated indicate that apoptotic pathways are of biological significance in pancreatic cancer. PMID- 9863490 TI - Prognostic factors in patients with endocrine tumours of the duodenopancreatic area. AB - BACKGROUND: The development of endocrine tumours of the duodenopancreatic area (ETDP) is thought to be slow, but their natural history is not well known. The aim of this study was to determine the factors that influence survival of patients with ETDP. PATIENTS/METHODS: Eighty two patients with ETDP (44 non functioning tumours, 23 gastrinomas, seven calcitonin-secreting tumours, four glucagonomas, three insulinomas, one somatostatinoma) followed from October 1991 to June 1997 were included in the study. The following factors were investigated: primary tumour size, hormonal clinical syndrome, liver metastases, lymph node metastases, extranodular/extrahepatic metastases, progression of liver metastases, local invasion, complete resection of the primary tumour, and degree of tumoral differentiation. The prognostic significance of these factors was investigated by uni- and multi-variate analysis. RESULTS: Twenty eight patients (34%) died within a median of 17 months (range 1-110) from diagnosis. Liver metastases (p = 0.001), lymph node metastases (p = 0.001), progression of liver metastases (p < 0.00001), lack of complete resection of the primary tumour (p = 0.001), extranodular/extrahepatic metastases (p = 0.001), local invasion (p = 0.001), primary tumour size > or = 3 cm (p = 0.001), non-functioning tumours (p = 0.02), and poor tumoral differentiation (p = 0.006) were associated with an unfavourable outcome by univariate analysis. Multivariate analysis identified only liver metastases (risk ratio (RR) = 8.3; p < 0.0001), poor tumoral cell differentiation (RR = 8.1; p = 0.0001), and lack of complete resection of the primary tumour (RR = 4.8; p = 0.0007) as independent risk factors. Five year survival rates were 40 and 100% in patients with and without liver metastases, 85 and 42% in patients with and without complete resection of primary tumour, and 17 and 71% in patients with poor and good tumour cell differentiation respectively. CONCLUSION: Liver metastases are a major prognostic factor in patients with ETDP. Progression of liver metastases is also an important factor which must be taken into account when deciding on the therapeutic approach. The only other independent prognostic factors are tumoral cell differentiation and complete resection of the primary tumour. PMID- 9863491 TI - Enhanced expression of monocyte tissue factor in patients with liver cirrhosis. AB - BACKGROUND: Previous studies have shown that cirrhotic patients produce increased amounts of thrombin but the underlying mechanism is still unknown. AIMS: To analyse the relation between the rate of thrombin generation and monocyte expression of tissue factor (TF) in cirrhosis. PATIENTS: Thirty three cirrhotic patients classified as having low (n = 7), moderate (n = 17), or severe (n = 9) liver failure according to Child-Pugh criteria. METHODS: Prothrombin fragment F1 + 2, monocyte TF activity and antigen, and endotoxaemia were measured in all patients. Polymerase chain reaction (PCR) analysis of TF mRNA was performed in monocytes of five cirrhotic patients. RESULTS: Prothrombin fragment F1 + 2 was higher in cirrhotic patients than in controls (p < 0.0001). Monocytes from cirrhotic patients had higher TF activity and antigen than those from controls (p < 0.001) with a progressive increase from low to severe liver failure. Monocyte expression of TF was significantly correlated with plasma levels of F1 + 2 (TF activity: r = 0.98, p < 0.0001; TF antigen: r = 0.95, p < 0.0001) and with endotoxaemia (TF activity: r = 0.94, p < 0.0001; TF antigen: r = 0.91, p < 0.0001). PCR analysis of TF mRNA showed TF expression only in three patients with endotoxaemia (more than 15 pg/ml). CONCLUSIONS: Cirrhotic patients have enhanced expression of TF which could be responsible for clotting activation, suggesting that endotoxaemia might play a pivotal role. PMID- 9863492 TI - Localisation and semiquantitative assessment of hepatic procollagen mRNA in primary biliary cirrhosis. AB - BACKGROUND: Chronic liver disease is characterised by excessive deposition of collagen and other extracellular matrix proteins, produced mainly, but not exclusively, by activated hepatic stellate cells in the perisinusoidal space. In primary biliary cirrhosis (PBC) fibrosis is concentrated mainly around the portal tracts. AIMS: To examine the hypothesis that, in addition to hepatic stellate cells, portal tract fibroblasts might play a significant role in the deposition of collagen in PBC. METHODS: Fifty liver biopsy specimens from patients with PBC were studied. An in situ hybridisation technique was adapted to localise and measure semiquantitatively type I procollagen mRNA in formalin fixed, paraffin wax embedded sections, using an 35S labelled cRNA probe specific for the alpha 1 chain of rat type I procollagen. Hepatic fibrogenic activity was also assessed using serum type III procollagen peptide (PIIINP). RESULTS: In PBC, type I procollagen gene expression was significantly increased. Signal was localised mainly in and around inflamed portal tracts, to cells which had the appearances of portal fibroblasts. Signal activity in these cells correlated with the degree of portal fibrosis and inflammation and also with serum PIIINP concentrations. CONCLUSIONS: Results are consistent with the hypothesis that the excessive extracellular matrix, deposited within the liver in PBC, is synthesised not only by hepatic stellate cells but also by portal tract fibroblasts. The semiquantitative assessment of procollagen mRNA in liver biopsy specimens may provide a useful method of evaluating the rate of synthesis of collagen and therefore disease activity in patients with PBC. PMID- 9863493 TI - H63D is an haemochromatosis associated allele. PMID- 9863494 TI - 22nd International Breast Cancer Research Congress of the International Association for Breast Cancer Research (IABCR). Athens, Greece, 24-27 September 1998. Abstracts. PMID- 9863495 TI - American Congress of Rehabilitation Medicine 75th anniversary. AB - Despite an explosive growth of rehabilitation programs for the disabled in the years following World War II, a national professional organization that could provide a forum for exchange of scientific information on an interdisciplinary basis about multiple disability concerns did not develop. Specialization of research, education, and practice in rehabilitation has increased markedly among basic scientists, engineers, nurses, occupational therapists, orthotists and prosthetists, physical therapists, physicians, psychologists, social workers, speech pathologists, and vocational counselors. This specialization... has caused many rehabilitation-related personnel to seek a central focus of professional interaction through interdisciplinary communication. The American Congress of Rehabilitation Medicine has undergone reorganization to invite multidisciplinary membership and participation in programs advancing the arts, sciences, and practices of rehabilitation medicine. This report reviews the methodology by which the reorganization was accomplished and records the progress made in restructuring membership, program, publications, and governance. The membership body is now multidisciplinary with full participation in the scientific forum on an interdisciplinary basis by those who are active in and contributing to the advance of medical rehabilitation services for the disabled. PMID- 9863496 TI - British Thoracic Society winter meeting. London, 2-4 December 1998. Abstracts. PMID- 9863497 TI - XVI National meeting of Clinical and Experimental Oncology, XXVI National symposium of the Italian Cancer Society, XXIV Scientific meeting of the Italian Association of Medical Oncology. Rome, 15-18 November 1998. Abstracts. PMID- 9863498 TI - Hypothetical reasons of the HIV1-AIDS "tritherapy" failure. A challenging model. AB - Daily journals in France recently published a declaration of eight AIDS assistance associations stating, because of already established resistance of most of types of HIV1 to the so called "tritherapies" 8,000 subjects in France will soon be "in condition of treatment failure". This tritherapeutic "flat note" is a double flat, for relative and absolute reasons: a) relative indeed was the case of well-known results: the tritherapies initially performed better than bitherapies, which had done better than monotherapies; b) absolute is their failure, which induce, as the other types, toxicity, resistance and relapses. Toxicity and resistance are due to the fact that, as the T1/2 of the virus is very short, virostatics must be applied continuously. But in AIDS groups tritherapies are applied not only in a continuous fashion, but in an identical form and for an undefined time, the process which is used in experimental cancer chemotherapy to induce resistant cell lines. Applying them in sequences of 3 weeks (a duration chosen with the knowledge that resistance may occur in about 12 weeks), we have shown in AIDS not only an absence of toxicity, but also an absence of resistance in patients treated with four drugs affecting four different targets. There is indeed another point to underline: AIDS group tritherapies are comprised of three drugs, but whatever the choices of these drugs they affect only two targets: retrotranscriptase and HIV1-protease. We had obtained in the best murine model of HIV1-infection (Friend's virus infection) eradication with a combination of three drugs, AZT, acriflavine (ACF) and the ellipticine analogue methyl-hydroxy-ellipticine (MHE); (the two last were discovered to be rather more efficient that AZT in our virostatic screening). This combination affects three virus targets (AZT, retrotranscriptase; MHE, topoisomerase 2; and ACF, integrated and proviral DNA). The next article will show that sequential drug combinations of three virostatics chosen from ten drugs available, affecting four targets, are more efficient in HIV1-AIDS than three drug combinations affecting three targets because they were chosen from a pool of only five drugs. It will, however, be shown that the same type of sequential combinations with four drug rotations chosen among the ten available ones affecting four targets rapidly reduced, and for years maintained, the viral load at undetectable levels. This level has been < 200 RNA copies/mL during the trial and is, at the end of the study, < 20 RNA copies/mL. PMID- 9863499 TI - Combinations of three or four HIV virostatics applied in short sequences which differ from each other by drug rotation. Preliminary results of the viral loads and CD4 numbers. AB - This paper presents the evolution during its follow-up of a virostatic combination study of the type I-II trial conducted on ten AIDS-related complex (ARC) or acquired immunodeficiency syndrome (AIDS) patients [1, 9, respectively]. Its concept is based on the following original notions: a) it is not the number of the virostatics applied to each patient at any phase which determines their effect; it is the number of affected virus targets which determines the effect. Thus, the so called "tritherapies", imposed by the "AIDS Command" to thousands of patients selected at random, to be compared to the same number of subjects receiving only "bi" or "monotherapies", might be beginning to face failure because they attack only two targets: retro-transcriptase and HIV1 protease. Having discovered, owing to our experimental screening, original HIV1 virostatics, acriflavine (ACF) and several ellipticine analogues among which we have used methyl-hydroxy-ellipticine (MHE), we are able to attack two virus targets unaffected by classical virostatics: ACF attacks DNA, from its integrated double branched stage to the provirus one, and MHE inhibits topoisomerase II. We experimentally combined these two agents with AZT, which inhibits retro transcriptase, thus we realized a combination affecting three targets. This three agent combination was able to eradicate Friend's virus from infected mice. Clinically, combinations of three drugs affecting four targets (as they are selected among the ten virostatics available today) give a stronger result than three drug combinations affecting only three targets, because they were selected from the five virostatics which were the only ones available at the beginning of the present study. Five patients out of five who received the combinations of four virostatics chosen among the ten currently available (thus affecting four targets) from the beginning of their treatment to the present have all reduced their viral load (VL) and maintained it below the detectable level (< 200 RNA copies/mL then 20 copies/mL); b) as the toxicities of virostatics and as HIV1 resistances may happen as soon as 12 weeks of treatment, the combinations have been, in our study, applied in shorter (3 week) sequences, differing from each other due to drug rotation; c) neither toxicity nor resistance occurred; d) curiously, the CD4 numbers, even when they increased rapidly, has never attained their normal count, and their curve may be a Gombertzian one. This CD4 restoration limitation can be due to persisting virus, as indicated in some patients by small peaks which may appear on some VL plateaus, though they disappear without treatment change. PMID- 9863500 TI - Potentialization of IL-2 effects on immune cells by oyster extract (JCOE) in normal and HIV-infected individuals. AB - Peripheral blood mononuclear cells (PBMCs) are physiologically activated by interleukin (IL)-2. We found that oyster extract (JCOE) currently used as a functional nutrient enhanced in vitro the IL-2 dependent activation as measured by cell count. 3H-thymidine uptake and up-regulation of a IL-2 receptor. In human immunodeficiency virus (HIV) seropositive individuals, this oyster extract induced effect was marked in asymptomatic individuals with quasi-normal CD4 cell counts, but was weakly reflected in acquired immunodeficiency syndrome (AIDS) patients. PMID- 9863501 TI - Phenylarsine oxide inhibits ex vivo HIV-1 expression. AB - Phenylarsine oxide (PAO), which is described as an inhibitor of tyrosine phosphatase activity, inhibits H2O2 release from human peripheral blood mononuclear cells (PBMCs) as measured by electrochemistry. Since human immunodeficiency virus type 1 (HIV-1) replication is known to be favored under oxidative stress conditions, ex vivo experiments using uninfected PBMCs, primary monocytes or a latently infected promonocytic U1 cell line show that HIV-1 replication and reactivation, monitored by p24 antigen measurement, are inhibited by PAO in a time- and concentration-dependent manner. These observations can be linked with the inhibition of NF-kappa B activation when uninfected monocytes are induced by either tumor necrosis factor alpha (TNF-alpha) phorbol 12-myristate 13 acetate (PMA) or lipopolysaccharide (LPS). PMID- 9863503 TI - Involvement of temperature sensitive syncytium inducing VSV or defective retroviruses in the development of spongiform encephalopathies. AB - Various enveloped viruses can induce syncytia in competent cells. Some temperature-sensitive mutants can express the trans-membrane viral G antigen under non-permissive conditions. The G antigen can then migrate at long distances, engulfing thousands of cells without producing any virus. When a temperature-sensitive vesicular stomatitis virus (VSV) infects a sensitive host, and under the condition that the G antigen is expressed, spongiosis can be induced in the central nervous system in the absence of detectable virus multiplication. We postulate that such a mechanism might be observed with various enveloped viruses, as recently illustrated with knock-out mice experimentally infected with defective murine leukemia virus (MULV). PMID- 9863502 TI - A controlled trial of dapsone versus pyrimethamine-sulfadoxine for primary prophylaxis of Pneumocystis carinii pneumonia and toxoplasmosis in patients with AIDS. AB - Pneumocystis carinii pneumonia (PCP) is the most common opportunistic human immunodeficiency virus (HIV)-related infection, occurring in 85% of HIV infected patients without prophylaxis. Preventive treatment is required when CD4 cell count falls below 200 cells per cubic millimeter. Cotrimoxazole has been shown to be highly effective but alternative drug regimens are often necessary because of the frequent drug hypersensitivity exhibited by HIV infected patients. The aim of this prospective, open, randomized, one-site study, involving HIV-infected patients with a CD4 cell count below 200/mm3, or a percentage under 20%, randomly assigned to receive either dapsone 50 mg daily or Fansidar one tablet weekly, was to compare the efficacy and safety of these drugs in the primary prophylaxis of PCP. Both dapsone and Fansidar appear to be safe and effective alternative agents for the prevention of PCP. Their role in Toxoplasma gondii prophylaxis requires further evaluation. PMID- 9863504 TI - Cerebral infarction associated with vasculitis due to varicella zoster virus in patients infected with the human immunodeficiency virus. AB - Cases of herpes zoster ophtalmicus (HZO) with delayed contralateral hemiparesis caused by hemispheric stroke secondary to granulomatous angiitis have been reported and are a well-recognized complication of herpes zoster. Similar cases have been reported more recently during infection with human immunodeficiency virus (HIV). We describe two HIV+ patients without any clinical history of zoster dermatitis who developed a sudden hemiparesis followed 2 weeks later for one by an acute retinal necrosis. Computerized tomography (CT) scan, magnetic resonance imaging (MRI), magnetic resonance angiography (MRA), and digital subtraction angiography (DSA) were performed and showed a hemispheric stroke with evidence of a segmental arteritis of the carotid syphon. Varicella zoster virus (VZV) was found in the cerebro spinal fluid (CSF) in the two patients and after puncture of the vitreous fluid of the patient with the acute retinal necrosis. These two cases exemplify the difficulty of diagnosis of stroke in HIV+ patients, which seems to be more frequent than in similarly aged non-infected patients and demonstrates that VZV needs to be taken in consideration and identified even without any past history of zoster dermatitis. PMID- 9863505 TI - Metabolic aspects of HIV: associated wasting. AB - Despite recent progress in the treatment of human immunodeficiency virus (HIV) infection, wasting syndrome (WS) is now one of the major aspects of acquired immunodeficiency syndrome (AIDS). Malnutrition in HIV infected patients is characterised by a predominant loss of body cell mass (BCM), the amount of functional protoplasm in non-adipose tissue. This loss of BCM is correlated with a higher risk of AIDS events and a greater risk of mortality. If anorexia plays a major role in the development of the WS, some abnormalities in the metabolism drive the predominant loss of BCM. In the stable state, the resting energy expenditure (REE) is increased by about 10%. The REE is significantly correlated with the whole body protein turn over as measured by C13 leucine. This particular and only metabolic situation is associated with an increased insulin sensitivity and a high level in de novo hepatic lipogenesis. During periods of secondary infections, patients had a striking average weight loss, resulting from the combination of anorexia and dramatic elevated REE. PMID- 9863506 TI - Two cases of cytomegalovirus infection revealed by hearing loss in HIV-infected patients. AB - Neurological complications are particularly common during HIV infection. Among various opportunistic diseases, cytomegalovirus (CMV) is one of the most frequent causes of central and peripheral neurological manifestations. Previously, there have been several reports of cranial nerve infection by CMV, but to our knowledge, no cases of auditory nerve involvement have been described. We report two cases in which CMV infection was revealed by involvement of the VIIIth cranial nerves. Cytomegalovirus (CMV) infection is frequent in severely immunodeficient patients infected by human immunodeficiency virus (HIV). The main targets of CMV are the retina, gastrointestinal tract and central nervous system. We describe two cases in which neurologic CMV infection was revealed by hearing loss. PMID- 9863507 TI - Anti-HIV activity of N-1-adamantyl-4-aminophthalimide. AB - The discovery of new leads acting via novel modes of action in the treatment of the human immunodeficiency virus (HIV), the causative agent of AIDS, remains a challenge. Along this line we synthesized and evaluated a series of N-substituted 4-aminophthalimides which were designed according to the models of thalidomide, phenytoin (PHT) and ameltolide. From a series of 24 compounds only N-1-adamantyl 4-aminophthalimide was endowed with anti-HIV-1 and -HIV-2 activity in CEM cell cultures. PMID- 9863508 TI - Frequencies of the CC chemokine receptor 5 delta 32 allele in various populations of defined racial background. AB - The 32-nucleotide deletion (delta 32) within the chemokine co-receptor CCR5 gene was studied in 1,031 DNA samples from various racial origins. The mutation is absent in 48 Vietnamese living in Paris, and we confirm that delta 32 allele is very rare in a sample of 302 individuals originating from sub-Saharan Africa. The mean delta 32 allelic frequency in unrelated non-HIV-1 infected blood donors is 0.113 in France, as calculated in a sample of 620 individuals. A slight decrease in allelic frequencies was noticed in the southern part of the country compared to the Paris region. In a population of 71 North African people the delta 32 frequency is 0.063, a value significantly lower than the mean frequency observed in France. PMID- 9863509 TI - Surgery in patients with HIV infection: indications and outcome. AB - We reviewed the records of 12 patients with HIV infection (one stage I, three stage II, two stage III, six stage IV) who received 15 surgical procedures under general or lumbar/epidural anesthesia. We discussed surgical indications, their poor wound healing and precautions for preventing the risk of transmission of HIV to health care workers. Six emergency and nine elective operations were performed. Postoperative complications developed after three emergency and three elective operations. Ten patients showed delay of wound healing which was not directly correlated with the CD4+ cell count. No operative deaths occurred. In any stage of HIV infection, not only palliative but also curative operations can be performed as long as HIV infection, opportunistic infections and HIV-related neoplasms can be controlled. Late stage wound healing is poor, but the wound will heal without keloid formation, although it takes two to three times longer than usual. For operating on patients with HIV infection precautions for preventing needle sticks, sharp injuries and blood exposure should be learned and used by health care workers. As a result, surgical staff members will be able to perform operations safely on HIV-infected patients to improve both quality of life and the prognosis of their disease. PMID- 9863510 TI - B-cell epitopes in HIV-1 Tat and Rev proteins colocalize with T-cell epitopes and with functional domains. AB - We describe the characterization of the B-cell epitopes of HIV-1 regulatory proteins Tat and Rev. The prevalence of antibodies to these proteins among human immunodeficiency virus (HIV)-1-infected individuals was examined by enzyme-linked immunosorbent assay (ELISA) and by Western blotting. The Tat and Rev antibody positive sera were selected for epitope mapping performed with partially overlapping synthetic peptides bound to polyethylene pins. Eighteen and twelve percent of HIV-infected individuals had antibodies against Tat or Rev, respectively. In Tat, four epitopic regions were identified, situated within amino acids 6-10 (PRLEP), 21-37 (ACTNCYCKKCCFHCQVC), 39-58 (ITKALGISYGRKKRRQRRRA) and 74-82 (TSQSRGDPT). The most frequently recognized epitopic regions were located in the middle of the protein. In Rev, the two most frequently recognized epitopic regions were near the amino terminus of the protein within amino acids 12-20 (LIRTVRLIK) and 38-49 (RRNRRRRWRERQ). A third epitope was mapped around amino acids 55-62 (ISERILGT) and a fourth around amino acids 78-83 (LERLTU). To analyze the specificity of Tat and Rev epitopes, soluble synthetic peptides representing the identified epitopes were used in an ELISA assay, and the recognition of most epitopes was shown to be specific for HIV-1-infected individuals. In addition, many of the Tat and Rev epitopes were shown to overlap with regions having functional activity or with regions previously identified as T-cell epitopes. PMID- 9863511 TI - Biomedical Engineering Society annual fall meeting. Cleveland, Ohio, USA. 10-13 October 1998. Abstracts. PMID- 9863513 TI - 146th Conference of the Society of Biochemistry and Molecular Biology, 6th photosynthesis workshop Nordwest. Bielefeld, October 10, 1997. Abstracts. PMID- 9863512 TI - 49th Mosbacher Colloquium. Molecular chaperones in biology and medicine. Mosbach, April 2-4, 1998. Abstracts. PMID- 9863514 TI - 156th Conference of the Society of Biochemistry and Molecular Biology, annual meeting of Study Group 3: Neurochemistry, main topic: Neuron-glia interaction. Tubingen, October 9-10, 1998. Abstracts. PMID- 9863515 TI - Society of Biochemistry and Molecular Biology fall meeting. Jena, September 27 30, 1998. Abstracts. PMID- 9863517 TI - 43rd Annual meeting of the German Society of Neuropathology and Neuroanatomy. Homburg, October 7-10, 1998. Abstracts. PMID- 9863516 TI - Epiperikaryal synaptophysin reactivity in the normal human central nervous system. AB - An epiperikaryal synaptophysin immunoreactivity has been regarded as an indicator of neoplastic or otherwise abnormal neurons and this staining property serves as an important criterion to distinguish between normal and abnormal neurons. In the present study we have investigated the epiperikaryal synaptophysin reactivity in various regions of the normal human central nervous system by using autopsy materials from 11 subjects aged 3 months-86 years. We found a definite but variable staining of the brainstem and spinal cord motor neurons as well as the cerebellar Purkinje cells. A particular strong and consistent reactivity was seen in neurons of the cerebellar nuclei in which also axons and dendrites were labelled to a variable extent. This type of neuronal staining was never observed in the cerebrum. We therefore conclude that the employment of this staining property as a criterion for abnormal neurons should be used with caution in the cerebellum, brainstem, and spinal cord whereas it may be used more safely in the cerebrum. PMID- 9863518 TI - XXth Congress of the European Society of Cardiology. Vienna, Austria, August 22 26, 1998. Abstracts. PMID- 9863519 TI - 29th Annual meeting of the German Society of Immunology. September 23-26, 1998. Abstracts. PMID- 9863520 TI - 5th International Symposium on Dendritic Cells in Fundamental and Clinical Immunology. Pittsburgh, Pennsylvania, USA. September 23-28, 1998. Abstracts. PMID- 9863521 TI - British Pharmaceutical Conference 1998, science proceedings 135th meeting. Eastbourne, September 8-11, 1998. Abstracts. PMID- 9863522 TI - The Physiological Society, proceedings of the Scientific meeting held at Charles University, Prague. 22-24 June 1998. Abstracts. PMID- 9863523 TI - 12th Annual North American Cystic Fibrosis Conference. Montreal, Quebec, Canada. October 15-18, 1998. Abstracts. PMID- 9863524 TI - Fluoride ion release from two methacrylate polymer systems. AB - The release of fluoride ions from two room-temperature polymerising systems containing sodium and potassium fluoride, respectively, has been studied. The polymer systems comprised poly(ethyl methacrylate) powder (PEM), with tetrahydrofurfuryl methacrylate (THFM), and n-butyl methacrylate (nBM), respectively. The water uptake of these systems was drastically increased by the presence of fluorides, the increase being much higher with the PEM/THFM system. In both cases, uptake was a monotonic function of the molarity of the fluoride added. The uptake process was in general non-Fickian. However, for all systems, the fluoride release process was Fickian, and diffusion coefficients could be calculated. The amount of fluoride released, and the diffusion coefficients obtained, appeared unrelated to the extent of water uptake of the parent polymers. PMID- 9863525 TI - Manufacturing and microscopical characterisation of polyurethane nerve guidance channel featuring a highly smooth internal surface. AB - The present study demonstrates the possibility of manufacturing polyurethane [ChronoFlex (CF)] nerve guidance channels (NGCs) featuring a highly smooth internal surface. Comparative SEM and AFM observations prove marked differences between the internal surface microgeometry of Silastic and CF channels. SEM of CF samples shows a surface with no detectable roughness, while Silastic channels show transversal rows along the entire surface. AFM digital image of Silastic samples show a surface with a rough microgeometry defined by a tridimensional pattern with peaks up to 1400 nm. AFM digital image of CF samples show, indeed, an essentially flat microgeometry with the highest level at 545 nm. These preliminary results suggest that the association of an innovative sequential deposition manufacturing technique with the new CF polyurethane may produce NGCs with a smoother surface microgeometry, in comparison to NGCs obtained from commercial Silastic tubes. PMID- 9863526 TI - Endothelial cells exposed to erythrocytes under shear stress: an in vitro study. AB - After injury and vascular replacement, endothelial cell recovery is limited and could lead to thrombosis. Seeding small diameter vascular prosthesis with endothelial cells has been proposed to fulfil cell lining and improve surface hemocompatibility. However, detachment of seeded cells occurs following implantation. Previous in vitro studies have looked at the fluid shear stress as a major cause of cell detachment. To our knowledge, the role of erythrocyte collisions has not been investigated. The present in vitro study aims at investigating whether endothelial cell adhesion depends on (i) the presence of erythrocytes in flow and (ii) the latent culture period (1, 24 and 48 h) between seeding and exposure to flow. Endothelial cells were exposed to culture media containing different erythrocyte concentrations using a steady laminar flow of 1350 ml min(-1) in a parallel plate flow chamber. Endothelial cell morphology in dynamic conditions was quantified and compared to that in static conditions. The projected area of cells were mostly found smaller under dynamic than static conditions, particularly at a wall shear stress of 23 dyn cm(-2). Cells from the 1 h latent culture period were oriented parallel to the flow axis and were more elongated than under static conditions. Conversely, endothelial cell shape was slightly modified when either the latent period or the wall shear stress was increased. Disparate orientation was observed on confluent endothelial cells (24 48 h latent period) exposed to shear stress with or without erythrocytes. Increasing fluid viscous forces due to erythrocytes play a critical role on the behaviour of freshly seeded endothelial cells upon exposure to blood flow. PMID- 9863527 TI - Hydroxyapatite fiber reinforced poly(alpha-hydroxy ester) foams for bone regeneration. AB - A process has been developed to manufacture biodegradable composite foams of poly(DL-lactic-co-glycolic acid) (PLGA) and hydroxyapatite short fibers for use in bone regeneration. The processing technique allows the manufacture of three dimensional foam scaffolds and involves the formation of a composite material consisting of a porogen material (either gelatin microspheres or salt particles) and hydroxyapatite short fibers embedded in a PLGA matrix. After the porogen is leached out, an open-cell composite foam remains which has a pore size and morphology defined by the porogen. By changing the weight fraction of the leachable component it was possible to produce composite foams with controlled porosities ranging from 0.47 +/- 0.02 to 0.85 +/- 0.01 (n = 3). Up to a polymer:fiber ratio of 7:6, short hydroxyapatite fibers served to reinforce low porosity PLGA foams manufactured using gelatin microspheres as a porogen. Foams with a compressive yield strength up to 2.82 +/- 0.63 MPa (n = 3) and a porosity of 0.47 +/- 0.02 (n = 3) were manufactured using a polymer:fiber weight ratio of 7:6. In contrast, high-porosity composite foams (up to 0.81 +/- 0.02, n = 3) suitable for cell seeding were not reinforced by the introduction of increasing quantities of hydroxyapatite short fibers. We were therefore able to manufacture high-porosity foams which may be seeded with cells but which have minimal compressive yield strength, or low porosity foams with enhanced osteoconductivity and compressive yield strength. PMID- 9863528 TI - Manufacture of porous biodegradable polymer conduits by an extrusion process for guided tissue regeneration. AB - We have fabricated porous, biodegradable tubular conduits for guided tissue regeneration using a combined solvent casting and extrusion technique. The biodegradable polymers used in this study were poly(DL-lactic-co-glycolic acid) (PLGA) and poly(L-lactic acid) (PLLA). A polymer/salt composite was first prepared by a solvent casting process. After drying, the composite was extruded to form a tubular construct. The salt particles in the construct were then leached out leaving a conduit with an open-pore structure. PLGA was studied as a model polymer to analyze the effects of salt weight fraction, salt particle size, and processing temperature on porosity and pore size of the extruded conduits. The porosity and pore size were found to increase with increasing salt weight fraction. Increasing the salt particle size increased the pore diameter but did not affect the porosity. High extrusion temperatures decreased the pore diameter without altering the porosity. Greater decrease in molecular weight was observed for conduits manufactured at higher temperatures. The mechanical properties of both PLGA and PLLA conduits were tested after degradation in vitro for up to 8 weeks. The modulus and failure strength of PLLA conduits were approximately 10 times higher than those of PLGA conduits. Failure strain was similar for both conduits. After degradation for 8 weeks, the molecular weights of the PLGA and PLLA conduits decreased to 38% and 43% of the initial values, respectively. However, both conduits maintained their shape and did not collapse. The PLGA also remained amorphous throughout the time course, while the crystallinity of PLLA increased from 5.2% to 11.5%. The potential of seeding the conduits with cells for transplantation or with biodegradable polymer microparticles for drug delivery was also tested with dyed microspheres. These porous tubular structures hold great promise for the regeneration of tissues which require tubular scaffolds such as peripheral nerve, long bone, intestine, or blood vessel. PMID- 9863529 TI - Novel hydrophilic cyclic monomers in hydrogel synthesis. AB - A range of hydrogels was prepared by the copolymerisation of 2-hydroxyethyl methacrylate (HEMA) with various hydrophilic cyclic monomers (CM) derived from cis-1,2-dihydroxy-3,5-cyclohexadiene (DHCD). The specific systems investigated were the cis-1,2-bis(2,3-epoxybutanoyloxy)-, cis-1,2-bis(10,11 epoxyundecanoyloxy)-, cis-1,2-bis(trimethylsiloxy)- and cis-1,2-bis(tert butyldimethylsiloxy)-3,5-cyclohexadienes. The EWC, tensile strength and initial modulus of the poly HEMA-CM hydrogels derived from the epoxy compounds showed lower EWCs but similar mechanical properties compared with polyHEMA itself. Treatment of the hydrogel derived from cis-1,2-bis(2,3-epoxybutanoyloxy)-3,5 cyclohexadiene with 20% sodium hydroxide increased the EWC value markedly but produced highly fragile materials. PMID- 9863530 TI - Tissue culture surface characteristics influence the expansion of human bone marrow cells. AB - Human cell therapy applications in tissue engineering, such as the ex vivo production of hematopoietic cells for transplantation, have recently entered the clinic. Although considerable effort has been focused on the development of biological processes to generate therapeutic cells, little has been published on the design and manufacture of devices for implementation of these processes in a robust and reproducible fashion at a clinical scale. In this study, the effect of tissue culture surface chemistry and texture was assessed in human bone marrow (BM) mononuclear cell (MNC) and CD34-enriched cell cultures. Growth and differentiation was assessed by total, progenitor (CFU-GM), stromal (CFU-F), and primitive (LTC-IC) cell output. Tissue culture treated (TCT) plastic significantly increased MNC culture output as compared with non-TCT plastic, whereas CD34-enriched cell cultures gave lower output (than MNC cultures) that was unaffected by TCT plastic. Interestingly, the level of MNC culture output was significantly different on four commercial TCT surfaces, with the best performing surface giving output that was 1.6- to 2.8-fold greater than the worst one. The surface giving the highest output was the best at supporting development of a distinct morphological feature in the adherent layer (i.e. cobblestone area) indicative of primitive cells, and X-ray photoelectron spectroscopy (XPS) was used to characterize this surface. For custom injection molding of culture devices, the use of three different resins resulted in MNC culture output that was equivalent to commercial cultureware controls, whereas CD34-enriched cell cultures were highly sensitive to resins containing additives. When the texture of molded parts was roughened by sandblasting of the tool, MNC culture output was significantly reduced and higher spikes of IL-6 and G-CSF production were observed, presumably due to macrophage activation. In conclusion, the manufacture of BM MNC culture devices for clinical applications was optimized by consideration of plastic resin, surface treatment, and texture of the culture substratum. Although CD34-enriched cells were insensitive to surface treatment, they were considerably more sensitive to biocompatibility issues related to resin selection. The development of robust systems for BM MNC expansion will enable clinical trials designed to test the safety and efficacy of cells produced in this novel tissue engineering application. PMID- 9863531 TI - Degradation of double-walled polymer microspheres of PLLA and P(CPP:SA)20:80. I. In vitro degradation. AB - Double-walled (DW) polymer microspheres with a core of poly(1,3-bis-(p carboxyphenoxy propane)-co-(sebacic anhydride)20:80 (P(CPP:SA)20:80) and an external coat of poly(L-lactic acid) (PLLA) were degraded for up to 6 months in vitro. The effects of hydrolytic degradation on the polymers were studied by Fourier-transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), gel permeation chromatography (GPC) and scanning electron microscopy (SEM). The inner core of the polyanhydride copolymer (P(CPP:SA)20:80) degraded first, breaking down into oligomers during the first week and then into monomers which were trapped by the outer shell of PLLA, crystallized, and remained in the core for the duration of the study. As expected, the PLLA coat degraded at a slower rate due to the ester bonds between lactic acid monomers being more hydrolytically resistant than the anhydride bonds. The PLLA in the DW microspheres decreased from its original molecular weight (Mw) of 24,000 g mol( 1) to approximately 5,000 g mol(-1) over the 6 month study. The melting temperature of the PLLA decreased more than 30 degrees C during the study, but no changes were observed in the FTIR spectra. The 6 month samples were very brittle, showing the concomitant drop in mechanical strength with the decrease in Mw. PMID- 9863532 TI - Degradation of double-walled polymer microspheres of PLLA and P(CPP:SA)20:80. II. In vivo degradation. AB - Double-walled (DW) polymer microspheres with a core of poly(1,3-bis-(p carboxyphenoxy propane)-co-(sebacic anhydride)20:80 (P(CPP:SA)20:80) and an external coat of poly(L-lactic acid) (PLLA) were implanted subcutaneously and intramuscularly and were allowed to degrade for up to 6 months. The tissue surrounding the implant was fixed and sectioned for histological evaluation by light and scanning electron microscopy. The remaining polymer was extracted with chloroform and analyzed by gel permeation chromatography (G PC), Fourier transform infrared (FTIR) spectroscopy, and differential scanning calorimetry (DSC). The inner core of the more hydrolytically labile P(CPP:SA)20:80 degraded first, becoming more granular over time. The PLLA coating showed no changes morphologically. The DW microspheres induced a stronger inflammatory response during the first week than did the SW PLLA microspheres, but this resolved by the second week. The SW PLLA microspheres showed no changes by SEM or optical microscopy. No differences in degradation were seen between samples implanted subcutaneously and those implanted intramuscularly. In vivo degradation was correlated to the results of an in vitro degradation study published as the first paper in this two paper series. PMID- 9863533 TI - The yielding, plastic flow, and fracture behavior of ultra-high molecular weight polyethylene used in total joint replacements. AB - The yielding, plastic flow, and fracture behavior of UHMWPE plays an important role in wear and failure mechanisms of total joint replacement components. The primary objective of this study was to compare the yielding, plastic flow, and fracture behavior of two implantable grades of UHMWPE (GUR 1120 vs 4150 HP). The first part of this work explored the hypothesis that up to the polymer yield point, the monotonic loading behavior of UHMWPE displays similar true stress strain behavior in tension and compression. Uniaxial tension and compression tests were conducted to compare the equivalent true stress vs strain response of UHMWPE up to 0.12 true strain. During monotonic loading, the equivalent true stress strain behavior was similar in tension and compression up to the yield point. However, investigation of the unloading behavior and permanent plastic deformations showed that classical deviatoric rate independent plasticity theory may dramatically overpredict the permanent strains in UHMWPE. A secondary goal of this study was to determine the ultimate true stress and strain for UHMWPE and to characterize the fracture surfaces after failure. Using a fracture mechanics approach, the critical flaw sizes were used in combination with the true ultimate stresses to predict the fracture toughness of the two resins. A custom video based strain measurement system was developed and validated to characterize the true stress-strain behavior up to failure and to verify the accuracy of the incompressibility assumption in calculating the true stress-strains up to failure. In a detailed uncertainty analysis, theoretical expressions were derived for the relative uncertainty in digital video-based estimates of nominal strain, true strain, homogeneous stress, and true stress. Although the yielding behavior of the two UHMWPE resins was similar, the hardening and plastic flow behavior clearly discriminated between the GUR 1120 and 4150 HP. A statistically significant difference between the fracture toughness of the two resins was also evident. The long-term goal of this research is to provide detailed true stress strain data for UHMWPE under uniaxial tension and compression for future numerical simulations and comparison with more complex multiaxial loading conditions. PMID- 9863534 TI - Oxidants and antioxidants in the pathogenesis of atherosclerosis: implications for the oxidized low density lipoprotein hypothesis. AB - The oxidation hypothesis proposes that low density lipoprotein must be oxidatively modified to trigger the pathological events of atherosclerosis. In this article, we evaluate recent studies addressing the pathways that promote low density lipoprotein oxidation in vivo and the impact of antioxidants on atherogenesis in animals, paying particular attention to the clinical implications of these studies for the oxidation hypothesis. PMID- 9863535 TI - Dyslipoproteinaemia and hyperoxidative stress in the pathogenesis of endothelial dysfunction in non-insulin dependent diabetes mellitus: an hypothesis. AB - Endothelial dysfunction in non-insulin dependent (Type 2) diabetes mellitus (NIDDM) has implications for the pathogenesis of the two major complications, macrovascular disease and microangiopathy. Endothelial dysfunction is a consequence of a disturbance in the L-arginine/nitric oxide pathway. Its occurrence in NIDDM is well supported by both in vitro and in vivo studies. NIDDM results in diverse abnormalities in lipoprotein metabolism, the most significant being hypertriglyceridaemia which is associated with increased plasma concentrations of small dense LDL and low levels of HDL. Dysglycaemia results in hyperoxidative stress and increased formation of advanced-glycosylation endproducts, both of which enhance the oxidative modification of lipoprotein particles. Based on extensive in vitro studies and on human data, we generate the hypothesis that the development of endothelial dysfunction in NIDDM is a consequence of the effect of dyslipoproteinaemia, in particular increased circulatory concentrations of modified small dense LDL and of hyperoxidative stress on the formation, action and disposal of nitric oxide, by diverse molecular mechanisms; HDL is proposed to have a protective effect on these processes through its enzymic antioxidant properties. The hypothesis proposed is simple, testable and consistent with wide sources of evidence. The practical implications of the hypothesis and the existing opportunities for the prevention and reversal of endothelial dysfunction in NIDDM are also reviewed and discussed. PMID- 9863536 TI - Dietary L-arginine and alpha-tocopherol reduce vascular oxidative stress and preserve endothelial function in hypercholesterolemic rabbits via different mechanisms. AB - Vascular oxidative stress brought about by superoxide radicals and oxidized low density lipoproteins (oxLDL) is a major factor contributing to decreased NO dependent vasodilator function in hypercholesterolemia and atherosclerosis. We investigated whether chronic administration of L-arginine (2% in drinking water) or of alpha-tocopherol (300 mg/day) improves endothelium-dependent vasodilator function and systemic NO production, reduces vascular oxidative stress, and reduces the progression of atherosclerosis in cholesterol-fed rabbits with pre existing hypercholesterolemia. Systemic NO production was assessed as urinary nitrate excretion; oxidative stress was measured by urinary 8-iso-PGF2alpha excretion in vivo, by lucigenin-enhanced chemiluminescence of isolated aortic rings ex vivo, and by copper-mediated LDL oxidation in vitro. Endothelium dependent relaxation was almost completely abrogated in cholesterol-fed rabbits. Urinary nitrate excretion was reduced by 46+/-10%, and 8-iso-PGF2alpha excretion was increased by 61+/-18% as compared to controls (each P <0.05). Vascular superoxide radical release stimulated by PMA ex vivo was increased by 273+/-93% in this group, and the lag time of LDL oxidation was reduced by 35+/-6% (each P <0.05). Treatment with L-arginine and alpha-tocopherol reduced intimal lesion formation (by 68+/-6 and 4+/-11%, respectively; P <0.05) and improved endothelium dependent relaxation. Both treatments also normalized urinary 8-iso-PGF2alpha excretion. L-Arginine increased urinary nitrate excretion by 43+/-13% (P <0.05) and reduced superoxide radical release by isolated aortic rings to control levels, which was unaffected by vitamin E treatment. By contrast, vitamin E dramatically increased the resistance of isolated LDL to copper-mediated oxidation in vitro by 178+/-7% (P <0.05), which was only marginally prolonged by L-arginine. Intimal thickening was reduced by both treatments. We conclude that both L-arginine and alpha-tocopherol reduce the progression of atherosclerotic plaques in cholesterol-fed rabbits. However, while L-arginine increases NO formation and reduces superoxide release, alpha-tocopherol antagonizes mainly oxLDL-related events in atherogenesis. Thus, both treatments reduce urinary isoprostane excretion and improve endothelium-dependent vasodilation via different mechanisms. PMID- 9863537 TI - Dual effects of the antioxidant agents probucol and carvedilol on proliferative and fatty lesions in hypercholesterolemic rabbits. AB - The in vivo direct antiatherogenic activity of the antioxidant probucol (200 mg/kg per day) or the beta-blocker with antioxidant properties carvedilol (10 and 20 mg/kg per day) was tested in the same animal in two different types of atherosclerotic lesion (proliferative and fatty lesions) induced in cholesterol fed rabbits (1%). Drugs were given daily mixed with standard diet for 8 weeks; body weight and plasma lipid profile were not different among groups throughout the study. Aortic fatty lesions were induced by cholesterol feeding (n = 25 in each group) and their extent expressed as % of aorta inner surface covered by plaques was significantly reduced by both drugs (28.2+/-9.6%, P <0.05, 19.9+/ 6.2%, P <0.01 for low- and high-dose carvedilol, respectively; 22.3+/-7.6%, P <0.01 for probucol, versus 41.6+/-10.7% in control rabbits). Proliferative lesions were obtained by positioning a hollow silastic collar around one carotid artery 6 weeks after dietary and drug treatments started (n = 5 in each group). The neointimal formation, mostly composed by myocytes, was determined by measuring cross-sectional thickness ratio of intimal (I) and medial (M) tissue of fixed arteries. In untreated animals, collared arteries resulted in a significant neointimal cell accumulation compared to the sham (1.10+/-0.14 versus 0.02+/ 0.01) without change in medial thickness. I/M ratio was reduced by about 50% in animals treated with probucol (0.51+/-0.1) and carvedilol (0.66+/-0.21 and 0.52+/ 0.1 in the low- and high-dose group, respectively). Total plasma TBARS were more than 50% lower in both probucol- and high-dose carvedilol-treated rabbits. Results show that pharmacological pretreatment with antioxidants directly inhibits early atherogenic processes, representing a potentially useful approach in the prevention of atherosclerosis. PMID- 9863538 TI - Oral administration of NO synthase inhibitor failed to promote arteriosclerotic lesions in the aorta and the coronary arteries of rabbits fed cholesterol. AB - We examined whether or not the oral administration of L-nitroarginine methylester (L-NAME), an inhibitor of nitric oxide (NO) synthase, promotes cholesterol induced arteriosclerosis in the aorta and the coronary artery. Thirty-six male Japanese white rabbits were fed 0.5% cholesterol-containing laboratory chow and randomly assigned to the following three groups: (1) water, (2) 80 microg/ml L NAME and (3) 400 microg/ml L-NAME in drinking water. The rabbits were fed a 0.5% cholesterol-containing diet for 8 months. During the 8-month period, the concentration of total cholesterol and L-nitroarginine in the serum and the mean blood pressure were measured. The concentration of NO3 in the serum was also measured. After sacrifice, the aortic surface involvement (AI%), the ratio of the thickened intima to the media and the contents of the total cholesterol of the aorta, the maximum % stenosis of the subepicardial large coronary artery, the % frequency of the nearly completely occlusive distal small coronary artery and the area of the myocardial fibrosis were all measured. We found no statistical difference among the three groups regarding the degree of arteriosclerotic lesions of the aorta and of the large coronary artery, and the area of myocardial fibrosis, as well as the serum cholesterol exposure index (the area under the curve of the serum total cholesterol concentration) and the mean blood pressure. However, the serum concentration of L-nitroarginine was approximately 50 and 200 microM/l in groups 2 and 3, respectively. The concentration of NO3 in the serum in group 1 was significantly higher than that in groups 2 and 3. We thus conclude, that the oral administration of L-NAME in the rabbits fed a cholesterol containing diet for 8 months failed to promote arteriosclerotic lesions in the aorta and the coronary artery, even though the serum concentration of L nitroarginine increased sufficiently to inhibit NO synthase in the arterial endothelium and the NO3 concentration in the serum decreased in the rabbits given L-NAME. PMID- 9863539 TI - Immunohistochemical localization of different epitopes of advanced glycation end products in human atherosclerotic lesions. AB - To better understand the role of advanced glycation end products (AGEs) in atherogenesis, we developed specific antibodies against different immunological epitopes of AGE structures, including Nepsilon-(carboxymethyl)lysine-protein adduct (CML) and a structure(s) other than CML (nonCML), and demonstrated the immunohistochemical localization of CML- and nonCML-epitopes in atherosclerotic lesions of human aorta, which were obtained at autopsy from 20 nondiabetic patients (12 males and eight females; mean age, 60.8+/-16.7 years). Monoclonal anti-CML antibody (6D12) recognized not only AGE-modified proteins, but also CML modified proteins. On the other hand, polyclonal anti-nonCML antibody reacted to AGE-modified proteins, but not to CML-modified proteins. Both antibodies were unreactive to the early-stage products of glycation, including fructose-modified butyloxycarbonyl-lysine and fructose-epsilon-aminocaproic acid. Atherosclerotic lesions included diffuse intimal thickening (DIT), fatty streaks (FS), atherosclerotic plaques (AP) and complicated lesions. An immunohistochemical analysis showed both CML- and nonCML-epitopes to be found along the collagen fibers in DIT in subjects more than 40 years old, but not in subjects less than 40 years old. CML-epitopes accumulated mainly in the cytoplasm of macrophage/foam cells, while nonCML-epitopes accumulated exclusively in the extracellular spaces in FS. APs showed the CML-epitope stored macrophage/foam cells, and the accumulation of both CML- and nonCML-epitopes in the lipid-rich fibrous area. An immunohistochemical analysis with a monoclonal antibody against oxidized low density lipoprotein (FOH1a/DLH3) showed the presence of this antigen within the cytoplasm of the macrophage/foam cells in atherosclerotic lesions, which were also positive for the CML-epitopes. These findings thus suggest that the heterogeneous localization of AGEs in atherosclerotic lesions depends on their different epitopes, and that a close link, therefore, exists between the peroxidation of LDL and the formation of AGEs in atherosclerotic lesions. PMID- 9863540 TI - High prevalence of small dense LDL in diabetic nephropathy is not directly associated with kidney damage: a possible role of postprandial lipemia. AB - To determine whether high prevalence of small dense low-density lipoprotein (LDL) in non-insulin-dependent diabetes (NIDDM) with nephropathy is directly associated with kidney damage, we measured LDL particle size by non-denaturing 2-16% gradient polyacrylamide gel electrophoresis in non-diabetic patients with primary renal disease and compared the results to particle size in NIDDM patients with diabetic nephropathy. The average LDL particle diameter was significantly smaller in patients with diabetic nephropathy (245+/-3 A mean +/- SEM) compared to the controls (263+/-1 A), diabetics without nephropathy (257+/-2 A), patients with primary renal disease (254+/-2 A) or non-diabetic patients treated with hemodialysis (HD) (260+/-1 A). The incidence of small LDL (mean diameter is < or =255 A) was remarkably increased in diabetic nephropathy (67%) compared to diabetes without nephropathy (27%), patients with renal disease (24%), HD patients (15%) and controls (10%). LDL size in patients with primary renal disease was significantly smaller than those in controls. However, because there was an excellent correlation between LDL size and plasma triglyceride (TG) levels, when hypertriglyceridemic subjects (TG >1.7 mM) were excluded, no difference of LDL size was observed between the renal disease group (260+/-2 A) and the control group (264+/-1 A). On the other hand, even when hypertriglyceridemic subjects were excluded, LDL size was still smaller in diabetic nephropathy (250+/-4 A). We performed an oral fat load test in normotriglyceridemic subjects (fasting TG <1.7 mM) of control, diabetes with and without nephropathy and primary renal disease. The TG responses in plasma and TG rich-lipoprotein (TRL) (d <1.006) after the oral fat load were significantly greater in NIDDMs with nephropathy compared to controls or NIDDMs without nephropathy, while such a marked postprandial lipemia was not observed in patients with primary renal disease. In these fasting normotriglyceridemic subjects, LDL size was significantly inversely correlated with postprandial TG responses, which is totally independent from fasting TG levels. These results suggest that high prevalence of small dense LDL in NIDDM patients with nephropathy is not directly associated with kidney damage. Postprandial lipemia may play an important role in reducing LDL particle size in these patients. PMID- 9863541 TI - Cholesteryl ester transfer in hypercholesterolaemia: fasting and postprandial studies with and without pravastatin. AB - Subjects with hypercholesterolaemia (HC) have increased fasting cholesteryl ester transfer protein (CETP) activity and accelerated cholesteryl ester transfer (CET) from HDL to apo B-containing lipoproteins. The aim of this study was to examine the effects of postprandial lipaemia and pravastatin treatment on plasma triglycerides (TG) and CETP activity and on CET and LDL Stokes' diameter in primary HC (n = 19, total cholesterol > or =6.5, LDL-cholesterol > or =4.5, TG <4.0 mmol/l). Samples were collected fasting and 6 h after an oral fat load (0.88 g/kg body weight) after 6 weeks therapy with placebo or pravastatin 40 mg nocte according to a double-blind randomized cross-over study. Apart from significant reductions in plasma total cholesterol, LDL-cholesterol apo B and TG. pravastatin significantly reduced CETP activity in both the fasting (mean +/- SD, 37.9+/-12.2 to 32.0+/-10.3 nmol/ml plasma per h) and postprandial state (35.5+/-11.3 to 31.3+/-9.5 nmol/ml plasma per h) compared to equivalent placebo phases. CETP activity did not change during postprandial lipaemia despite a significant 45-55% increase in CET to triglyceride-rich lipoproteins (TRL) of d <1.006 g/ml. LDL Stokes' diameter was unchanged postprandially or by pravastatin. The mass of TRL was the strongest contributor to variation in CET in both fasting and postprandial plasma, accounting for at least 77% of the variance of CET. Postprandial TRL-TG was the strongest contributor to variation in fasting LDL Stokes' diameter in untreated HC (54%) whilst HDL-cholesterol was the strongest fasting contributor to variation (45%) for placebo- and pravastatin-treated HC. We conclude that pravastatin may reduce the atherogenicity of the lipoprotein profile in HC by reducing CETP activity. Furthermore, CET is strongly influenced by postprandial lipaemia which may have a cumulative effect on LDL size. PMID- 9863542 TI - LPS-induced cytokine production and expression of beta2-integrins and CD14 by peripheral blood mononuclear cells of patients with homozygous familial hypercholesterolemia. AB - It has been suggested that proinflammatory cytokines such as tumor necrosis factor-alpha (TNF) and interleukin-1beta (IL-1), as well as adhesion molecules such as beta2-integrins and CD14, play a role in the pathogenesis of atherosclerosis. Familial hypercholesterolemia (FH) is an autosomal disease in which defective or absent LDL receptors are the cause for extreme LDL concentrations and early development of atherosclerosis. We studied lipopolysaccharide-induced cytokine production and the expression of adhesion molecules by mononuclear cells of three homozygous FH patients and compared them with first-degree relatives and healthy controls. There was a tendency towards increased cytokine production by cells of FH patients, whereas the expression of adhesion molecules was not modified compared to controls. In addition, LDL apheresis inhibited IL-1 and TNF production and the expression of CD11a, CD11b, CD11c and CD14 by the mononuclear cells of FH patients and this may be an additional beneficial effect of LDL-apheresis apart of decreasing LDL concentrations. PMID- 9863543 TI - Association of malondialdehyde-acetaldehyde (MAA) adducted proteins with atherosclerotic-induced vascular inflammatory injury. AB - Atherosclerosis is a vascular injury characterized by elevated tissue levels of tumor necrosis factor-alpha (TNF-alpha), increased expression of endothelial cell adhesion molecules, and vascular wall inflammatory cell infiltration. Foam cells are associated with atherosclerotic plaque material, and low density lipoprotein (LDL) is a lipid component of foam cells. Malondialdehyde (MDA) is an oxidative product of unsaturated fatty acids and is also present in atherosclerotic lesions. MDA-modified (adducted) proteins, including MDA-modified LDL, are present in atherosclerotic human vascular tissue. Acetaldehyde (AA) is the major metabolic product of ethanol oxidation. Both MDA and AA are highly reactive aldehydes and will combine with proteins to produce an antigenically distinct protein adduct, termed the MAA adduct. This study demonstrates that proteins modified in the presence of high concentrations of MDA can produce MAA-modified proteins in vitro. In addition, MAA adducted proteins are capable of inducing rat heart endothelial cell cultures (rHEC) to produce and release TNF-alpha, and cause rHEC upregulation of endothelial adhesion molecule expression, including ICAM-1. These adhesion molecules are required for circulating inflammatory cells to adhere to endothelium which allows inflammatory cell tissue infiltration. Additionally, MAA modified proteins were defected in human atherosclerotic aortic vascular tissue but not in normal aortic tissue. Since atherosclerosis is associated with an inflammatory vascular injury characterized by elevated tissue TNF-alpha concentrations and inflammatory cell infiltration, these data suggest that MAA-adducted proteins may be formed in atherosclerotic plaque material and may be involved in the inflammatory reaction that occurs in atherosclerosis. These data further suggest that previous studies demonstrating MDA modified protein in atherosclerotic plaque may in fact have MAA modified proteins associated with them. PMID- 9863544 TI - Raman spectroscopy for quantifying cholesterol in intact coronary artery wall. AB - The chemical composition of vascular lesions, an important determinant of plaque progression and rupture, can not presently be determined in vivo. Prior studies have shown that Raman spectroscopy can accurately quantify the amounts of major lipid classes and calcium salts in homogenized coronary artery tissue. This study determines how the relative cholesterol content, which is calculated from Raman spectra collected at the luminal surface of an artery, is related to its depth in an intact arterial wall. Raman spectra of human atherosclerotic plaques were measured after thin tissue layers were successively placed on them. From these spectra, relative cholesterol contents were calculated and used to determine how cholesterol signal strength is attenuated by overlaying tissue. Then, intact artery samples (n = 13) were examined spectroscopically, sectioned and stained specifically for cholesterol. Images of these sections were digitized, and image intensities were related to cholesterol content. These cholesterol amounts were weighed appropriately for depth into the tissue and area-integrated for comparison with spectroscopy results. A decaying exponential curve was fit to the layer study data (r2 = 0.97) and showed that approximately 300 microm of tissue attenuates cholesterol signals by 50%. In intact plaques, the spectroscopically determined cholesterol amounts correlated strongly and linearly with those determined by digital microscopy (r2 = 0.94). With Raman spectroscopy techniques, the cholesterol content of a lesion can be determined by properly accounting for its depth into an arterial wall. Our results suggest that chemical concentrations in an artery wall could be mapped throughout its thickness, possibly by combining Raman spectroscopy methods with other techniques. PMID- 9863545 TI - In vitro growth suppression of vascular smooth muscle cells using adenovirus mediated gene transfer of a truncated form of fibroblast growth factor receptor. AB - Vascular smooth muscle cell (VSMC) proliferation associated with arterial injury causes restenosis, which remains to be resolved in cardiovascular and ischemic cerebrovascular disease, especially after balloon angioplasty. Fibroblast growth factor (FGF) is a potent mitogen and a trophic factor for a variety of cells, including VSMCs. We constructed a replication-deficient adenovirus vector, designated AxCA delta FR, coding a truncated form of fibroblast growth factor receptor-1 (FGFR-1) gene lacking the intracellular domain to interrupt receptor mediated FGF signaling, and examined its effect on the proliferation of primary cultured rat VSMCs. We transferred the truncated form of the FGFR-1 gene to the VSMCs and confirmed its expression and localization in infected cells by Western blotting and immunofluorescence study. The VSMCs infected with AxCA delta FR degenerated and the proliferation of these cells was suppressed markedly by the infection with this virus in vitro. Our results suggest that the receptor mediated signal of FGFs has an important role in VSMC proliferation and gene transfer of a truncated form of FGFR using adenoviral vector may be useful for the treatment of the diseases caused by excessive proliferation of VSMCs like restenosis after percutaneous transluminal angioplasty or carotid endoarterectomy. PMID- 9863546 TI - The influence of acute smoking on leucocytes, platelets and the endothelium. AB - Cigarette smoking is a risk factor for the development of atherosclerosis. Possible mechanisms for this include leucocytes and platelet activation, and/or damage to the endothelium, any of which may contribute to changes in thrombosis and haemostasis. We examined the acute effects of smoking on these systems by obtaining blood before, immediately after, and at 10 and 30 min after the rapid smoking of two cigarettes in sequence by 20 smokers. Blood samples taken at the same time points from ten non-smokers acted as control material. In the smokers there was a transient rise in leucocyte count and neutrophil activation, but von Willebrand factor (VWF--marking endothelial damage) increased steadily at each time point (P <0.05). There were no changes in neutrophil elastase, soluble intercellular adhesion molecule-1 (sICAM-1 normally increased in smokers), fibrinogen, platelet count or soluble P-selectin (marking platelet activation, also normally increased in smokers). We conclude that the acute smoking of two cigarettes in succession will activate leucocytes and cause endothelial cell damage, but will not immediately influence platelet activity. PMID- 9863547 TI - Inhibition of endothelial cell adhesion and proliferation by extracellular matrix from vascular smooth muscle cells: role of type V collagen. AB - Endothelial cells recovering from damage due to disease or surgical procedures come into close contact with extracellular matrix (ECM) secreted by intimal vascular smooth muscle cells (VSMCs). We have investigated these relationships using human umbilical artery endothelial cells (HUAECs) and human mammary artery VSMC in vitro. HUAEC adhesion and proliferation were significantly lower on ECM secreted by VSMC compared with HUAEC ECM or surface-coated fibronectin. Characterisation of the ECM of both cell types with monoclonal antibodies showed that the ECM secreted by VSMC contained significantly more elastin, chondroitin sulphate and collagen types I, III and V than that from HUAECs. HUAECs adhered poorly to collagen type V coated on plastic and not at all to elastin. When these proteins were co-coated with fibronectin, elastin did not inhibit migration or proliferation compared to the response on fibronectin but collagen type V significantly inhibited both. Treatment of VSMC ECM with enzymes which selectively depleted the matrix of collagen types I, III and IV, or chondroitin sulphate, had no effect on HUAEC responses to the ECM, suggesting that these molecules did not contribute to the inhibition of HUAECs. Treatment of VSMC ECM with a mixture of collagenases, selectively depleted the matrix of collagen type V, as well as types I, III and IV. Such depleted ECMs supported increased proliferation of HUAECs compared to buffer controls. Overall these results suggest that collagen V secreted into the ECM of VSMC may inhibit the recovery of adjacent endothelium. PMID- 9863548 TI - A description of two morphologic patterns of aortic fatty streaks, and a hypothesis of their pathogenesis. AB - Two morphologic patterns of fatty streak were identified on examination of 74 aortas from the Pathobiological Determinants of Atherosclerosis in Youth study. Pattern 1, which predominated in 78% of aortas, is characterized by broad bands of intense stain which extend to the proximal edge of ostia. Pattern 2, which predominated in 11%, is characterized by less intense staining which is concave to the associated ostium. Pattern 1 predominated in older subjects and smokers. Aging and smoking decrease arterial elasticity, thereby decreasing the volume and duration of retrograde blood flow in diastole. Doppler ultrasonography of the posterior intercostal arteries and aorta in 42 healthy subjects revealed that retrograde blood flow in late systole/early diastole is normal in subjects in the 15-34 age group. Transition from retrograde to antegrade flow was associated with transient blood stasis. This stasis should prolong the residence time of lipid rich particles, enhancing diffusion into the vessel wall. A region of lower flow velocity was noted in the periostial region in all patients during diastole. The anatomic, hemodynamic, and risk factor data suggest that the morphology of fatty streaks is determined by interaction of retrograde with antegrade blood flow as modulated by arterial elasticity. PMID- 9863549 TI - The effect of a common methylenetetrahydrofolate reductase mutation on levels of homocysteine, folate, vitamin B12 and on the risk of premature atherosclerosis. AB - An increased total plasma homocysteine level is an established risk factor for atherosclerotic vascular disease. The plasma level of homocysteine is influenced by both environmental and genetic factors. An important genetic determinant of plasma homocysteine is a common amino acid dimorphism (Ala222Val) in the methylenetetrahydrofolate reductase (MTHFR) gene. Individuals homozygous for the Val allele have significantly higher homocysteine levels than those with an Ala/Val or Ala/Ala genotype. Moreover, the Val/Val genotype has been claimed to be a strong genetic risk factor for atherosclerosis. The aim of the present study is: (1) to determine the risk associated with the MTHFR dimorphism by comparing the genotype distribution in patients with premature atherosclerosis with that in a group of healthy controls; and (2) to investigate the relationship between the MTHFR genotype and parameters of homocysteine metabolism. The patient group consisted of 257 consecutive referred individuals with angiographically proven premature ( <50 years of age) arterial disease (coronary, and/or peripheral vascular disease). A total of 272 healthy hospital workers without a history of vascular disease were selected as a control group. The MTHFR-genotype was determined by PCR and gel-electrophoresis. A methionine-loading test was performed on 245 patients, and, in addition to homocysteine, levels of folate and vitamin B12 were measured. We found a strong correlation between MTHFR genotype and plasma homocysteine levels both before and after methionine loading. In addition, the MTHFR genotype seems important for the inverse relationship between homocysteine and folate and vitamin B12 levels. Lastly, the MTHFR genotype distribution was not different between patient and control groups. MTHFR genotype is a strong determinant of plasma homocysteine levels. Moreover, the plasma level of folate, which by itself influences homocysteine levels, is also dependent on the MTHFR genotype. In Val/Val genotypes, low levels of both folate and B12 lead to a relatively large increase in homocysteine levels. Nevertheless, the MTHFR genotype does not increase the risk for premature coronary artery disease. PMID- 9863550 TI - Association of the insertion/deletion gene polymorphism of the apolipoprotein B signal peptide with myocardial infarction. AB - The Del allele of the apolipoprotein B (apoB) signal peptide (SP) insertion/deletion (Ins/Del) polymorphism has been shown to be associated with elevated plasma levels of apoB, cholesterol and low density lipoprotein. It was the aim of the present study to analyse the relation of this gene variation to the risk of coronary artery disease (CAD) and of myocardial infarction (MI) in a population of 2259 male Caucasians, whose coronary anatomy was defined by means of coronary angiography. ApoB SP DelDel genotypes had significantly higher apoB plasma concentrations than InsIns homozygotes (P = 0.0001) and InsDel heterozygotes (P = 0.002); however, the apoB plasma levels of InsIns and InsDel genotypes were essentially the same (P = 0.54). Similar observations were made with respect to ApoB SP genotype-dependent cholesterol plasma concentrations. Since the apoB plasma level was not only associated with the apoB SP Ins/Del gene variation but also to the extent of coronary artery disease (P <0.0001), individuals with an InsIns genotype and without CAD had the lowest and subjects with a DelDel genotype and triple vessel disease the highest apoB plasma levels (P <0.0001). An association of the apoB SP Ins/Del gene variation with CAD was not detected, neither in the total population nor in low risk groups. In contrast, the gene variation was associated with MI (P <0.05). An Odds ratio of 1.18 (95% CI, 1.01-1.39) associated with the Del allele was detected in the total sample (P <0.02). In a subpopulation of individuals with low plasma triglyceride levels ( <154 mg/dl; mean value) and an DD genotype of the angiotensin I converting enzyme insertion/deletion gene polymorphism an Odds ratio of 2.01 (1.42-3.05) was calculated (P <0.001). The present study presents evidence for a statistically significant difference in the development of MI between genotype classes of the apoB SP Ins/Del gene polymorphism. PMID- 9863551 TI - Raised levels of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in the plasma of patients with ischaemic heart disease and hyperlipidaemia. PMID- 9863553 TI - Importance of metabolism in pharmacological studies: possible in vitro predictability. AB - Metabolic transformation of drug leads to the formation of a large number of secondary compounds. These metabolites may (a) participate to the elimination of the patent drug, (b) have similar or different therapeutic effects compared to the parent drug (c) exert toxic effects. Cytochromes P450 are the main enzymes involved in the biotransformation of exogenous drugs, leading to oxidized, reduced or peroxidized metabolites. Different isozymes of P450 are present in already all the organs and differ by their affinity for substrate families. P450 3A is the most abundant P450 protein in the adult human liver and is able to transform hundreds of substrates into either drugs or endogenous compounds such as testosterone. Its catalytic activities are regulated either by induction or by inhibition. Attempts to predict metabolic transformation of a given drug are based on the amount of P450 expressed in heterologous systems, induction, and inhibition experiments and by comparison to classical P450 substrates. Erythromycin metabolism and its P450 effects are used to illustrate the complexity and the consequences of metabolic transformation of a given drug. PMID- 9863552 TI - Radio-imaging for quantitative autoradiography in biology. AB - We present here an overview of new in vitro and ex vivo radio-imaging systems developed to overcome the limitations of films and emulsions currently used in histological autoradiography experiments. The shortcomings of films for quantitative studies are first introduced. Principles and performances of each family of imagers are discussed and illustrated in various biological contexts. Finally, perspectives of development including nonradioactive labeling techniques are briefly presented. PMID- 9863554 TI - Isolated working heart: description of models relevant to radioisotopic and pharmacological assessments. AB - Isolated heart preparations are used to study physiological and metabolic parameters of the heart independently of its environment. Several preparations of isolated perfused heart are currently used, mainly the retrograde perfusion system and the working heart model. Both models allow investigations of the metabolic regulation of the heart in various physiological conditions (changes in workload, hormonal influences, substrate competition). These systems may also reproduce different pathological conditions, such as ischemia, reperfusion and hypoxia. Quantitation of metabolic activity can be performed with specific radioactive tracers. Finally, the effects of various drugs on cardiac performance and resistance to ischemia can be studied as well. Heart perfusion also revealed efficient methods to determine the tracer/tracee relation for radioisotopic analogues used with Positron Emission Tomography. PMID- 9863555 TI - Whole hemisphere autoradiography of the postmortem human brain. AB - Whole hemisphere autoradiography (WHA) with selective high-affinity radioligands is a new tool in studying the distribution of receptors and of other neuronal components postmortem in brains from controls vs. subjects with psychiatric and neurologic diseases. WHA can be performed with several different isotopes (e.g., 3H, 125I, and 11C), and is in addition to characterization studies also used as a tool in early radioligand development. Moreover, using this technique, high resolution images are obtained that are complementary to those obtained in vivo with e.g., PET and SPECT. Results on dopamine and serotonin receptor subtypes as well as of their transporters show that WHA is a very suitable technique for the detailed characterization of the distribution in the whole human brain. PMID- 9863556 TI - Modeling dopamine system dysfunction in experimental animals. AB - Quite a substantial number of human disorders have been associated with a primary or a secondary impairment of one or several of the dopaminergic pathways. Among disorders associated with a primary impairment of dopaminergic transmission are Parkinson's disease, striatonigral degeneration, progressive supranuclear palsy, and possibly schizophrenia. Diseases of secondary dopamine dysfunction are chiefly represented by Huntington's disease in which dopaminergic transmission is being interrupted by progressive loss of the striatal neurons bearing the postsynaptic D1- and D2-dopamine receptors. Central dopaminergic systems have anatomical as well as organizational properties that render them unique by comparison to other neurotransmission systems, making them able to play a pivotal role in the modulation of various important brain functions such as locomotor activity, attention, and some cognitive abilities. These properties of dopamine neurons have obviously several implications in the clinical expression of human disorders involving dopamine neuron dysfunction. In addition, they can greatly influence the clinical/behavioral consequences of experimental lesions in animal models of dopamine dysfunctions. PMID- 9863557 TI - Positron emission tomography (PET) methodology for small animals and its application in radiopharmaceutical preclinical investigation. AB - The use and usefulness of positron emission tomography (PET) to quantify the specific and selective in vivo binding of radioligands in small laboratory animals is briefly reviewed up to the end of 1996. Emphasis is placed on practical experience with a dedicated, small diameter, tomograph (built in collaboration with CTI, Knoxville, TN), implementing conventional PET methodology. PMID- 9863558 TI - Preclinical evaluation of radiopharmaceutical toxicological prerequisites. AB - Radiopharmaceuticals should be tested in a battery of assays to evaluate human risk assessment, as for every drug. The battery of tests used for this evaluation should be adapted taking into account the specificity of radiopharmaceuticals and particularly the short duration of treatment, the toxicity associated both to the unlabelled part of the molecule and to the radioactivity itself, but also to a specific type of impurities due to the radiolytic degradation products. PMID- 9863559 TI - In vivo radiolabel quantification in small-animal models. AB - Current developments in emission tomography especially designed for small-animal imaging are presented. Adaptations of the human tomography principles take advantage of the smaller field of view to achieve about 2-mm usable resolution. Some evaluations in rat tomography are presented, and the problems of limiting resolution of PET and SPECT systems are discussed. Finally, a new approach that more specifically takes into account the parameters induced by in vivo quantification in rodents is presented. PMID- 9863560 TI - Functional neuroanatomy of the basal ganglia as studied by dual-probe microdialysis. AB - Dual probe microdialysis was employed in intact rat brain to investigate the effect of intrastriatal perfusion with selective dopamine D1 and D2 receptor agonists and with c-fos antisense oligonucleotide on (a) local GABA release in the striatum; (b) the internal segment of the globus pallidus and the substantia nigra pars reticulata, which is the output site of the strionigral GABA pathway; and (c) the external segment of the globus pallidus, which is the output site of the striopallidal GABA pathway. The data provide functional in vivo evidence for a selective dopamine D1 receptor-mediated activation of the direct strionigral GABA pathway and a selective dopamine D2 receptor inhibition of the indirect striopallidal GABA pathway and provides a neuronal substrate for parallel processing in the basal ganglia regulation of motor function. Taken together, these findings offer new therapeutic strategies for the treatment of dopamine linked disorders such as Parkinson's disease, Huntington's disease, and schizophrenia. PMID- 9863561 TI - Animal models for studying serotonin (5-HT) receptor subtypes: relationship to 5 HT system pathologies. AB - Despite more than 30 years of intensive research, the specific role of serotonin (5-HT) receptor subtypes in animal models of "anxiety" still remains unclear. The present study focused on the particular role of 5-HT1A receptor subtype in aversive learning, i.e., the passive avoidance (PA) task in the rat. Taken together, the data strongly suggest that: (1) 5-HT1A receptor but not 5-HT2A receptors play a crucial role in PA; (2) the postsynaptic but not presynaptic 5 HT1A receptors are mainly involved in the regulation of PA; (3) 5-HT1A receptors appear to be directly involved both in acquisition and retrieval but not in the consolidation of PA; and (4) besides the "prototypical" 5-HT1A receptor subtype, an additional and yet-unidentified 5-HT receptor subtype seems to play an important modulatory role in PA. PMID- 9863563 TI - N-[11C]methylpiperidine esters as acetylcholinesterase substrates: an in vivo structure-reactivity study. AB - A series of simple esters incorporating the N-[11C]methylpiperidine structure were examined as in vivo substrates for acetylcholinesterase in mouse brain. 4-N [11C]Methylpiperidinyl esters, including the acetate, propionate and isobutyrate esters, are good in vivo substrates for mammalian cholinesterases. Introduction of a methyl group at the 4-position of the 4-piperidinol esters, to form the ester of a teritary alcohol, effectively blocks enzymatic action. Methylation of 4-N-[11C]methylpiperidinyl propionate at the 3-position gives a derivative with increased in vivo reactivity toward acetylcholinesterase. Esters of piperidinecarboxylic acids (nipecotic, isonipecotic and pipecolinic acid ethyl esters) are not hydrolyzed by acetylcholinesterase in vivo, nor do they act as in vivo inhibitors of the enzyme. This study has identified simple methods to both increase and decrease the in vivo reactivity of piperidinyl esters toward acetylcholinesterase. PMID- 9863562 TI - Synthesis of 1-[11C]methylpiperidin-4-yl propionate ([11C]PMP) for in vivo measurements of acetylcholinesterase activity. AB - Synthesis of 1-[11C]methylpiperidin-4-yl propionate ([11C]PMP), an in vivo substrate for acetylcholinesterase, is reported. An improved preparation of 4 piperidinyl propionate (PHP), the immediate precursor for radiolabeling, was accomplished in three steps from 4-hydroxypiperidine by (a) protection of the amine as the benzyl carbamate, (b) acylation with propionyl chloride, and (c) deprotection of the carbamate by catalytic hydrogenation. The final product was obtained in an overall 82% yield. Reaction of the free base form of PHP with [11C]methyl trifluoromethanesulfonate at room temperature in N,N dimethylformamide, followed by high performance liquid chromatography (HPLC) purification, provided [11C]PMP in 57% radiochemical yield, > 99% radiochemical purity, and > 1500 Ci/mmol at the end of synthesis. The total synthesis time from end-of-bombardment was 35 min. [11C]PMP can thus be reliably prepared for routine clinical studies of acetylcholinesterase in human brain using positron emission tomography. PMID- 9863564 TI - Syntheses of carbon-11 labeled piperidine esters as potential in vivo substrates for acetylcholinesterase. AB - A series of carbon-11 labeled N-methylpiperidinyl esters were prepared as potential in vivo substrates for acetylcholinesterase (AChE). Target compounds were designed based on the structure of N-[11C]methylpiperidin-4-yl propionate, an ester currently used to measure AChE enzymatic activity in the human brain, to examine the structure-activity relationship for in vivo enzymatic hydrolysis. Changes in steric bulk and in the ester order ("reverse" esters) were made. Addition of methyl groups was made to both the acid side chain (synthesis of N [11C]methylmethylpiperidin-4-yl isobutyrate) and to the piperidine ring (syntheses of N-[11C]methyl-4-methylpiperidin-4-yl propionate, N-[11C]methyl-4 methylpiperidin-4-yl acetate, and N-[11C]methyl-3-methylpiperidin-4-yl propionate). Alterations of the order of the ester heteroatoms was accomplished through syntheses of the N-[11C]methyl-2,3- and 4-piperidinecarboxylic acid ethyl esters. Finally, an additional piperidine-based ester (N-[11C]methylpiperidin-2 yl)methyl propionate was also prepared. All carbon-11-labeled esters were prepared by N-[11C]methylation reactions, using the desmethyl precursors and no carrier-added [11C]methyltriflate, and were obtained in decay-corrected yields (not optimized) of 10-40% and high specific activities. PMID- 9863566 TI - Influence of acetylcholine on binding of 4-[125I]iododexetimide to muscarinic brain receptors. AB - The distribution of nicotinic and muscarinic cholinergic receptors in the human brain in vivo has been successfully characterized using radiolabeled tracers and emission tomography. The effect of acetylcholine release into the synaptic cleft on receptor binding of these tracers has not yet been investigated. The present study examined the influence of acetylcholine on binding of 4 [125I]iododexetimide to muscarinic cholinergic receptors of porcine brain synaptosomes in vitro. 4-Iododexetimide is a subtype-unspecific muscarinic receptor antagonist with high affinity. Acetylcholine competed with 4 [125I]iododexetimide in a dose-dependent manner. A concentration of 500 microM acetylcholine inhibited 50% of total specific 4-[125I]iododexetimide binding to synaptosomes when both substances were given simultaneously. An 800 microM acetylcholine solution reduced total specific 4-[125I]iododexetimide binding by about 35%, when acetylcholine was given 60 min after incubation of synaptosomes with 4-[125I]iododexetimide. Variations in the synaptic acetylcholine concentration might influence muscarinic cholinergic receptor imaging in vivo using 4-[123I]iododexetimide. Conversely, 4-[123I]iododexetimide might be an appropriate molecule to investigate alterations of acetylcholine release into the synaptic cleft in vivo using single photon emission computed tomography. PMID- 9863565 TI - Derivatives of WAY 100635 as potential imaging agents for 5-HT1A receptors: syntheses, radiosyntheses, and in vitro and in vivo evaluation. AB - Analogues of the potent and selective 5-HT1A ligand, WAY 100635, were synthesized and examined as potential candidates for imaging 5-HT1A receptors by positron emission tomography (PET). Several of the analogues displayed nanomolar affinity for the 5-HT1A receptor, comparable to WAY 100635. Three of these were examined in a model of human liver metabolism vis-a-vis WAY 100635. All showed a markedly lower propensity for amide hydrolysis than WAY 100635. Radiolabelling of these three potential PET radiotracers with carbon-11 was readily achieved from [11C] iodomethane, and the newly synthesized radioligands were tested in vivo in rats for binding to 5-HT1A receptors. Whereas two of the ligands failed to bind to 5 HT1A receptors in vivo, one was successful. The latter, [11C]-7 [4-(2' methoxyphenyl)-1-[2'-[N-(2'-pyridinyl)-2-bicyclo[2.2.2]octanec arboxamido]ethyl] piperazine], showed good brain penetration, hippocampal:cerebellar ratios of 10:1 at 45 min postinjection. Blocking studies with a variety of drugs demonstrated that the binding of [11C]-7 in vivo was selective for 5-HT1A receptors. [11C]-7 is a promising candidate as a ligand for imaging 5-HT1A receptors by PET. PMID- 9863567 TI - New iodinated progestins as potential ligands for progesterone receptor imaging in breast cancer. Part 1: Synthesis and in vitro pharmacological characterization. AB - Five putative iodinated progesterone receptor (PR) binding ligands were synthesized and evaluated as potential imaging agents for PR-positive human breast tumours. Two compounds (E- and Z-17-hydroxy-21-iodo-19-nor-17alpha-pregna 4,20-dien-3-one; E- and Z-IPG1) were previously described, but are re-evaluated. The other three were novel compounds: two nortestosterone analogues derived from ORG 3236 (E- and Z-13-ethyl-17-hydroxy-21-iodo-11-methylene-18,19-dinor-17alpha pre gna-4,20-diene-3-one; E- and Z-IPG2) and one norprogesterone analogue derived from ORG 2058 (21-[4-iodophenoxy]-16alpha-ethyl-19-norpregn-4-ene-3, 20-dione; IPG3). The E-iodovinyl nortestosterone compounds were obtained by a new route of synthesis. Competitive binding studies were performed to determine their binding affinities for the PR in three types of tissue (human MCF-7 breast tumour cells and rat uterine and mammary tumour tissue) and for the androgen receptor (AR) in human MCF-7 breast tumour cells, as well as for the sex hormone-binding globulin (SHBG) and corticosteroid-binding globulin (CBG) in human plasma. All four 17alpha-iodovinyl nortestosterone derivatives displayed high binding affinity for the human PR, that of Z-IPG1 and E- and Z-IPG2 being even higher than that of ORG2058. Their affinities for the rat PR were somewhat lower, especially those of both E-isomers. The affinity of IPG3 was lower for both the human and rat PR. The nortestosterone derivatives also showed AR binding, the relative binding affinities ranging from 4.3 to 17.0% as compared with 5alphaDHT. Additionally, neither of these steroids displayed any significant binding to either SHBG or CBG in human plasma. We conclude that the in vitro binding properties of all four 17alpha-iodovinyl nortestosterone derivatives warrant evaluation of the distribution characteristics of their 123I-labelled analogues to determine their usefulness as PR imaging agents. PMID- 9863568 TI - New iodinated progestins as potential ligands for progesterone receptor imaging in breast cancer. Part 2: In vivo pharmacological characterization. AB - On the basis of the observed high selective binding to both the human and rat progesterone receptor (PR) in vitro, three 17alpha-iodovinyl-substituted nortestosterone derivatives, i.e., the Z-isomer of 17alpha-iodovinyl-19 nortestosterone (Z-IVNT; Z-IPG1) and both the stereoisomers of 17alpha-iodovinyl 18-methyl-11-methylene-19-nortestosterone (E- and Z-IPG2), were selected for radio-iodination and subsequently evaluated as potential radioligands for PR imaging in human breast cancer. Their target tissue uptake, retention, and uptake selectivity were studied in female rats. The distribution studies revealed that PR-mediated uptake in the uterus and ovaries could only be demonstrated for Z [123I]IPG2. The target tissue uptake selectivity was, however, low, with the highest uterus-to-nontarget tissue uptake ratios observed at 2-4 h postinjection (p.i.), being 4.4, 1.8, and 7.4 for the uterus-to-blood, -fat, and -muscle ratio, respectively. For Z-[123I]IPG2, distribution was also studied in dimethylbenzanthracene (DMBA)-induced mammary tumour-bearing rats and in normal rabbits. Mammary tumour uptake of Z-[123I]IPG2 in the mammary tumour-bearing rat was also found to be PR-specific. In rabbits, higher selective target tissue uptake of Z-[123I]IPG2 was observed than in rats, resulting in uterus-to-blood, fat, and -muscle ratios of 6.6, 2.2, and 21.3 at 2-4 h p.i., respectively. In conclusion, Z-[123I]IPG2, which displayed high binding affinity for both the human and rat PR in vitro, showed specific PR-mediated target tissue uptake in rats and rabbits in vivo, the uptake selectivity being highest in the latter. Because the binding characteristics appeared to vary between species, a pilot study in breast cancer patients may be needed to decide whether Z-[123I]IPG2 can be of potential use as PR imaging agent in breast cancer. PMID- 9863569 TI - Radiotracer-derived biochemical profiles via a mathematical model: implications for therapeutic intervention. PMID- 9863570 TI - Recommendations of a Task Force of the European Society of Cardiology and the European Resuscitation Council on The Pre-hospital Management of Acute Heart Attacks. PMID- 9863571 TI - The influence of chest compressions and external defibrillation on the release of creatine kinase-MB and cardiac troponin T in patients resuscitated from out-of hospital cardiac arrest. AB - OBJECTIVES: This study sought to determine the influence of resuscitative procedures, such as chest compressions and external defibrillation, on the release of creatine kinase (CK)-MB and cardiac troponin T (cTnT). METHODS: In 87 patients with out-of-hospital cardiac arrest and successful cardiopulmonary resuscitation (CPR), the initial ECG rhythm, the duration of cardiac arrest and CPR, and the number of defibrillations were assessed on arrival in the hospital. The serum CK-MB and cTnT were measured 12 h after the event. We also assessed whether the patient developed cardiogenic shock within 12 h, and if the patient had acute myocardial infarction (AMI), which was confirmed or eliminated by of typical ECG findings, thallium-201 myocardial scintigraphy, or autopsy within the hospital stay. A backward stepwise linear regression model was applied to assess the association between the markers of myocardial injury (CK-MB and cTnT) and the above clinical variables. RESULTS: CK-MB concentrations were independently associated with the presence of AMI [B 68.5 (SE 28.5, P = 0.018)], the duration of CPR (as a measure of trauma to the chest by means of chest compressions) [B 2.07 (SE 1.01, P = 0.045)] and cardiogenic shock [B 52.3 (SE 23.4, P = 0.03)]. The remaining clinical variables listed were excluded by the model. Cardiac troponin T concentrations were only associated with the presence of AMI [B 4.86 (SE 1.34, P = 0.0005)]. There was a non-significant association between increasing serum cTnT concentrations and the presence of cardiogenic shock [B 2.51 (SE 1.46, P = 0.09)]. The remaining clinical variables were excluded by the model. CONCLUSION: The release of CK-MB appears to be influenced by the duration of resuscitation and the presence of cardiogenic shock. This has to be considered when interpreting serum CK-MB concentrations after CPR. The release of cTnT seems to be only associated with acute myocardial infarction, but not with the duration of chest compressions, or with the number of defibrillations administered. PMID- 9863573 TI - Respiratory system compliance decreases after cardiopulmonary resuscitation and stomach inflation: impact of large and small tidal volumes on calculated peak airway pressure. AB - The purpose of the present study was to evaluate respiratory system compliance after cardiopulmonary resuscitation (CPR) and subsequent stomach inflation. Further, we calculated peak airway pressure according to the different tidal volume recommendations of the European Resuscitation Council (7.5 ml/kg) and the American Heart Association (15 ml/kg) for ventilation of an unintubated cardiac arrest victim. After 4 min of ventricular fibrillation, and 6 min of CPR, return of spontaneous circulation (ROSC) after defibrillation occurred in seven pigs. Respiratory system compliance was measured at prearrest, after ROSC, and after 2 and 4 l of stomach inflation in the postresuscitation phase; peak airway pressure was subsequently calculated. Before cardiac arrest the mean (+/- S.D.) respiratory system compliance was 30 +/- 3 ml/cm H2O, and decreased significantly (P < 0.05) after ROSC to 24 +/- 5 ml/cm H2O, and further declined significantly to 18 +/- 4 ml/cm H2O after 2 l, and to 13 +/- 3 ml/cm H2O after 4 l of stomach inflation. At prearrest, the mean +/- S.D. calculated peak airway pressure according to European versus American guidelines was 9 +/- 1 versus 18 +/- 3 cm H2O, after ROSC 12 +/- 2 versus 23 +/- 4 cm H2O, and 15 +/- 2 versus 30 +/- 5 cm H2O after 2 l, and 22 +/- 6 versus 44 +/- 12 cm H2O after 4 l of stomach inflation. In conclusion, respiratory system compliance decreased significantly after CPR and subsequent induction of stomach inflation in an animal model with a wide open airway. This may have a significant impact on peak airway pressure and distribution of gas during ventilation of an unintubated patient with cardiac arrest. PMID- 9863572 TI - Myocardial perfusion during cardiopulmonary resuscitation (CPR): effects of 10, 25 and 50% coronary stenoses. AB - Myocardial blood flow under normal physiologic conditions remains relatively unaltered by coronary lesions with diameter stenosis of less than 70%. However, during cardiac arrest and cardiopulmonary resuscitation (CPR), autoregulation of coronary blood flow is compromised. We studied the effect of previously considered 'trivial' coronary stenoses of 10, 25 and 50% on distal myocardial perfusion in 39 swine undergoing CPR for cardiac arrest. Endocardial blood flow distal to the stenosis was significantly less than that proximal in all the different stenoses groups (P < 0.05). A 50% diameter stenosis also compromised epicardial blood flow distal to the stenosis (P < 0.05). This study suggests that any coronary lesion may compromise myocardial perfusion during CPR. Since the vast majority of cardiac arrest victims have coronary artery disease, careful attention to maximizing myocardial blood flow during CPR is needed. PMID- 9863574 TI - Anomalous ECG downloads from semi-automatic external defibrillators. AB - The coincidental print-out by two different Laerdal systems (subsequently called 'system A' and 'system B') of the same medical control module (MCM) for a Laerdal Heartstart 2000 semi-automatic external defibrillator (SAED) led to the discovery of three deficiencies in the information storage and printing processes. First, we noted that the impedance reported via system A was consistently higher. Second, we found the attachment of 'mysterious' ECG samples in the reports from system B, but not from system A. A third problem was the unpredictable (in)ability of system B to print out the information from the MCMs. Further investigations with help from the company suggested that the above-mentioned problems were caused by incompatibilities between the software in the different parts of equipment used (i.e. SAED devices, MCMs, printing systems and a computer program to store the information in a database). These observations demonstrate the need for strict medical supervision on all aspects of a SAED project, and for feed-back from clinicians to manufacturers. PMID- 9863575 TI - Measuring the effectiveness of contact tracing. PMID- 9863576 TI - HIV infection and pregnancy. PMID- 9863577 TI - The outcome of contact tracing for gonorrhoea in the United Kingdom. AB - Tracing and treating contacts (partner notification, PN) is a crucial part of the control of sexually transmitted infections but few studies have quantified its outcome. A retrospective national study obtained information from 155 clinics on 1308 patients with gonorrhoea, 59% of all UK patients attending in January to March 1995. These gave 1887 contacts (1:1.5) of whom 621 attended, 75% being found to be infected. The ratio of contacts attending to index patients was 0.5 (621/1260). The ratio was not influenced by gender or ethnicity, but was affected by orientation, being 0.5 for contacts of male heterosexuals, 0.3 for contacts of male homosexuals (P <0.001). PN outcome was less good in metropolitan (0.3) than provincial clinics (0.5), suggesting that further research is necessary on the reasons for this difference. Reliable ongoing outcome audit will require good communication between clinics to verify that contacts have attended. The contact:index ratio is suitable for evaluation of PN as it relates to actual outcome rather than to the patient's account of contact numbers. Local work can be assessed against these national figures. PMID- 9863578 TI - PCR testing of genital and urine specimens compared with culture for the diagnosis of chlamydial infection in men and women. AB - Our aim was to determine the number of chlamydial infections detected by Cobas Amplicor CT/NG multiplex polymerase chain reaction (PCR) testing of genital and first-voided urine (FVU) specimens compared with routine culture. Two hundred and eighty-six female and 276 male patients attending the Genito-Urinary Medicine (GUM) Unit at Edinburgh Royal Infirmary were included in the study. Case notes were analysed retrospectively to determine how many infected patients would not have been treated had diagnosis relied on routine culture. Polymerase chain reaction on FVU from women had a sensitivity, specificity, positive and negative predictive value of 91%, 100%, 100% and 99.1%: corresponding values for genital PCR and culture were 96%, 100%, 100%, 99.6% and 65%, 100%, 100%, 96.7% respectively. PCR on FVU from men had a sensitivity, specificity, positive and negative predictive value of 96%, 99.1%, 92.6% and 99.5%: corresponding values for genital PCR and culture were 89%, 99.5%, 95.8%, 98.6% and 48%, 100%, 100%, 94.3% respectively. In both men and women genital PCR and urine PCR were significantly more sensitive than culture. PCR almost doubled the number of patients detected by culture (49 vs 27). Of the 22 cases detected only by PCR 8 would not have received treatment on the basis of clinic treatment policy. PMID- 9863579 TI - Sexual histories, partnerships and networks associated with the transmission of gonorrhoea. AB - We aimed to describe and compare sexual links among people with gonorrhoea, by studying patients in 2 UK departments of genitourinary medicine. Interviews were completed for 510 and 235 cases in London and Sheffield respectively. There was a greater proportion of cases in men, homosexual men, non-white and non-British people and fewer female sex workers in London. Total networks of 1738 people in London and 570 people in Sheffield were described. Large linked heterosexual networks identified in Sheffield were associated with local contact, including men with higher numbers of sexual partners. Condom use for vaginal sex was reported for 11% of heterosexual partnerships in Sheffield, and 27% in London, with little difference between regular and casual partners. It was more difficult to define networks in London due to a high proportion of relatively anonymous contacts. These difficulties suggest that research and interventions may profitably focus upon venues for meeting partners as well as partner notification. PMID- 9863580 TI - Sexual functioning in ambulatory men with HIV/AIDS. AB - Sexual functioning is often neglected in the care of HIV-infected patients. Little information exists about the relationship between hormonal factors, psychological factors, medication, HIV disease stage and sexual functioning among persons with HIV disease. In this study, 50 HIV+ men completed the Derogatis Sexual Functioning Inventory (DSFI), and had serum hormonal assays drawn (testosterone, thyroid function test, leuteinizing hormone, prolactin and oestradiol). Although all the subjects reported some degree of sexual dysfunction, persons with symptomatic HIV/AIDS reported more negative mood, lower sexual satisfaction scores and worse body image than persons with asymptomatic HIV. Persons with asymptomatic HIV also tended to have normal testosterone levels compared with persons with symptomatic HIV/AIDS. No relationship was found between medications and low testosterone, although numbers were small. These results suggest that sexual dysfunction is prevalent among persons with HIV disease, is more common as patients become symptomatic and progress to AIDS and that both physiological (low testosterone) and psychological issues play a role. PMID- 9863581 TI - Gynaecological conditions associated with HIV infection in women who are partners of HIV-positive Thai blood donors. AB - Women who were partners of HIV-positive blood donors were enrolled in a study of heterosexual HIV transmission between March 1992 and December 1996 and were interviewed and examined. Gynaecological conditions, including cervical dysplasia, human papillomavirus (HPV) infection, gonorrhoea, chlamydial infection, trichomoniasis, bacterial vaginosis, vaginal candidiasis and syphilis were assessed in addition to HIV status and CD4 level. Of 481 women enrolled, 224 (46.6%) were HIV seropositive. HIV-infected women were more likely to have abnormal vaginal discharge on physical examination (OR=2.6, P <0.01), HPV infection with a high-risk type (OR=6.9, P <0.01), and cervical dysplasia (OR=5.3, P <0.01). The prevalence of other gynaecological conditions detected at the enrolment visit did not differ by HIV status. History of prior STD (OR=2.0, P <0.01) was more common among HIV-infected women. The median CD4 count was 400 cells/microl among HIV-infected women. The prevalence of abnormal vaginal discharge and bacterial vaginosis increased significantly with decreasing CD4 count. The prevalence of ectopy, vaginal candidiasis, and cervical dysplasia increased with decreasing CD4 count, but these trends were not significant. We conclude that HIV-infected Thai women appear to have increased prevalences of abnormal vaginal discharge, squamous intraepithelial lesions and self-reported history of STD. PMID- 9863582 TI - Mannose-binding protein in HIV-seropositive patients does not contribute to disease progression or bacterial infections. AB - This study set out to investigate whether plasma mannose-binding protein (MBP) deficiency caused by mutations in the MBP gene associates with pyogenic or opportunistic infections in HIV-infected patients. Plasma samples were selected randomly from 131 HIV-infected patients followed prospectively for a period not exceeding 12 months or until death. Plasma MBP concentrations were measured by an ELISA and genotyping was determined by amplification of exon 1 of the MBP gene by polymerase chain reaction (PCR) technology, followed by restriction enzyme analysis and Southern blotting using sequence-specific oligonucleotide probes. Neither MBP concentration nor genotype was found to associate with disease progression or opportunistic infection rate. There was an unexpected increased bacterial infection rate in patients with MBP levels greater than 100 ng/ml and wild type genotype. Thus, MBP does not appear to play a role in HIV infection. MBP is an acute phase reactant and this may explain the higher levels in those with more frequent pyogenic infections. PMID- 9863583 TI - An assessment of care provided by a public sector STD clinic in Cape Town. AB - A study was undertaken in a Cape Town public sector STD clinic to evaluate the content and quality of care provided since it has been recognized that appropriate improvements in the management of conventional sexually transmitted diseases (STDs), including provision of correct therapy, health education, condom promotion and partner notification, could result in a reduced incidence of HIV infection. Our objectives were to assess patients' needs for health education and to assess the quality of STD management in terms of health education, condom promotion, partner notification, the validity of the clinical diagnoses and the adequacy of the treatments prescribed. The study subjects were sampled systematically, according to their gender. Patients included in the study were given a standardized interview and their clinical records reviewed. Specimens were collected for laboratory investigations. For each STD detected, the treatment was defined as adequate if drugs currently known to be active against that infection were prescribed. One hundred and seventy men and 161 women were included in the study (median age: females 22 years, males 26 years). While almost all patients believed their STD may have been caused by unprotected sexual intercourse, many also believed it may have been caused by other factors, such as bewitchment with traditional medicine. Only 21% of male and 37% of female patients received any education about STD transmission during the clinic visit, and only 25% of male and 36% of female patients received education about condom use. As a result of the low sensitivity of the clinicians' diagnoses, 16% of men and 61% of women left the clinic with at least one infection inadequately treated. The majority of patients were not receiving education for the prevention of STDs including HIV. Many were not receiving adequate treatment for their infections. The introduction of a syndromic management protocol in this setting would substantially reduce the proportion of inadequately-treated patients. However, syndromic protocols, and the means by which they are implemented, need to take into account problems with the clinical detection of genital ulcerative disease and candidiasis in women. PMID- 9863584 TI - Views of Cameroonian sex workers on a woman-controlled method of contraception and disease protection. AB - Five hundred and twenty female Cameroonian sex workers participating in a clinical trial of the effectiveness of nonoxynol-9 (N-9) contraceptive film to prevent HIV infection were interviewed to determine their experience with this woman-controlled method. The sex workers liked the film, with over 80% stating they would continue to use it if it were shown to be effective against HIV and were to become widely available. However, they were much more likely to use condoms and films simultaneously with their clients (40%) than with their regular partners (16%). The advantages ascribed to the film were that its use was under the woman's control; it could be used without the partner's knowledge; it allowed direct contact between partners; and it provided back-up protection in case of condom breakage. Data from this small study suggest that development of an effective, woman-controlled method of pregnancy and disease protection would be welcomed by these women. PMID- 9863585 TI - A comparison audit of a genitourinary medicine colposcopy service against nationally agreed quality standards. AB - The National Health Service Cervical Screening Programme (NHSCSP) recently published their document Standards and Quality in Colposcopy outlining recommendations for standards in the service provided by colposcopists. In this first audit of colposcopic practice measured against these agreed UK national guidelines, the authors discuss the standards, their attainability, and their relevance to genitourinary medicine (GUM) colposcopy services. PMID- 9863586 TI - Genital ulcers in a primary health clinic in Rwanda: impact of HIV infection on diagnosis and ulcer healing (1986-1992). AB - During 1986-88 and 1990-92, 1025 (97%) out of 1057 genital ulcer patients in Kigali, Rwanda, were tested for HIV antibodies and for infection with Treponema pallidum, Haemophilus ducreyi and herpes simplex virus. Overall, 57% of men and 80% of women had antibodies to HIV-1. The most frequent laboratory diagnoses were chancroid (27%), syphilis (19%) and genital herpes (19%) among men and syphilis (35%), genital herpes (23%) and chancroid (20%) among women. HIV-1 seroprevalence increased sharply over time among men but not among women. The clinical presentation of ulcers as well as laboratory diagnoses were similar in the HIV-1 seropositive and seronegative groups. The relative frequency of all laboratory diagnoses remained unchanged over time. HIV-1 seropositivity had no impact on ulcer healing. Advanced immunodeficiency was diagnosed among 12% of the HIV-1 seropositive patients and was significantly associated with increasing age and genital herpes. PMID- 9863587 TI - Coronary artery disease occurring with protease inhibitor therapy. PMID- 9863588 TI - Outcome of inflammatory smears managed in a genitourinary medicine clinic. PMID- 9863589 TI - Soluble intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) in patients with HIV/AIDS does not appear to correlate with cytomegalovirus retinitis. PMID- 9863590 TI - Mutations in the TSC1 gene account for a minority of patients with tuberous sclerosis. AB - Tuberous sclerosis (TSC) is an autosomal dominant disorder characterised by tumour-like malformations (hamartomas) of the brain, skin, and other organs, often associated with seizures and learning disability. There is genetic heterogeneity with loci for TSC on chromosomes 9q34 (TSC1) and 16p13.3 (TSC2). The recently cloned TSC1 gene has 23 exons spanning some 40 kb of genomic DNA with an 8.6 kb transcript. We now report the results of mutation screening by SSCP and heteroduplex analysis of genomic DNA for all 21 coding exons of TSC1 in 83 unrelated cases of tuberous sclerosis. TSC1 gene mutations were found in 16 of the 83 cases (19%). These comprised base substitutions, small insertions, or small deletions giving rise to six nonsense mutations, eight frameshifts, and two splice site mutations, all of which would be expected to result in a truncated or absent protein. In the 10 cases predicted to have TSC1 mutations by linkage analysis or loss of heterozygosity studies, the mutation was identified in eight (80%). In the remaining 73 unassigned cases, only eight mutations were found (11%). From these data we estimate that TSC1 mutations accounted for 24% of the cases in this sample (and an estimated 22% of all TSC cases). This contrasts with data from linkage studies suggesting that TSC1 and TSC2 mutations account for approximately equal numbers of families. PMID- 9863591 TI - Differential diagnosis of type 2 neurofibromatosis: molecular discrimination of NF2 and sporadic vestibular schwannomas. AB - Patients who present with unilateral vestibular schwannomas either at a young age or with additional features of type 2 neurofibromatosis (NF2) are at risk of developing bilateral disease and transmitting a risk of neurogenic tumours to their offspring. We have identified 15 patients from a series of 537 with unilateral vestibular schwannomas who also had one or more of the following: other tumours (10/15), features of NF2 (3/15), or a family history of neurogenic tumours (5/15). No germline NF2 mutations were detected and in 7/9 cases where tumour material was available for analysis a germline mutation in the NF2 gene has been excluded. Although a possibility of gonosomal mosaicism still exists, exclusion tests for the offspring are now possible. We suggest a general strategy, based on analysis of tumour DNA, for distinguishing sporadic and familial cases of tumours caused by two hit mechanisms. Application of this strategy suggests that most instances of unilateral vestibular schwannoma which do not fulfil criteria for NF2 represent chance occurrences. PMID- 9863592 TI - Genetic implications of double primary cancers of the colorectum and endometrium. AB - Hereditary non-polyposis colorectal cancer (HNPCC) is an autosomal dominant condition predisposing to cancers of the colorectum and endometrium. Endometrial cancer is the most commonly occurring extracolonic cancer in HNPCC. Estimates of the cumulative incidence of endometrial cancer in women with mutations in the HNPCC genes range from 22-43%. In order to determine how frequently double primary cancers of the colorectum and endometrium are the result of a hereditary factor, we conducted a registry based study in Ontario and Quebec, Canada. We obtained pedigrees on 80 women diagnosed with double primary cancers of the colorectum and endometrium at less than 70 years of age. Family histories of cancer were obtained for all first degree relatives of these women and cancer rates were compared with age standardised provincial incidence rates in order to estimate the relative risks. There was a total of 82 cancers observed in relatives below the age of 55, compared with 31.2 expected, giving a relative risk of 2.6 (95% confidence interval (CI) 2.1-3.3). The relative risk for colorectal cancer below 55 was 16.1 (95% CI 11.6-21.8). This risk decreased with increasing age of onset of cancers in probands. For probands with both colorectal and endometrial cancer diagnosed under the age of 55, the relative risk of colorectal cancer in relatives below the age of 55 was 30.5 (95% CI 18.8-46.6). Similar patterns were observed for endometrial and pancreatic cancer. There were non-significant increases in rates of cancer of the oesophagus, stomach, small intestine, and bladder. There was no increased risk of breast cancer. The risk of lung cancer was decreased, especially in older relatives. Our findings indicate the presence of a significant genetic component of cancer in women with double primary cancers of the colorectum and endometrium. PMID- 9863593 TI - Locus heterogeneity in autosomal dominant congenital external ophthalmoplegia (CFEOM). AB - Congenital external ophthalmoplegia (CFEOM) is an uncommon autosomal dominant condition that has previously been mapped to the pericentromeric region of chromosome 12 in seven families with no evidence of locus heterogeneity. We report three families with typical CFEOM. One family does not map to this region of chromosome 12 or to other chromosomal locations implicated in disorders of lid or ocular movement. Recombinants in two CFEOM families potentially help to reduce the size of the candidate region on chromosome 12. PMID- 9863594 TI - Novel TIGR/MYOC mutations in families with juvenile onset primary open angle glaucoma. AB - Mutations in the trabecular meshwork induced glucocorticoid response protein (TIGR) or myocilin (MYOC) has recently been shown to cause juvenile onset primary open angle glaucoma (JOAG). In this study, we identified two new mutations (Asp380Ala and Ser502Pro) in two British families and another (Pro370Leu) in a French-Canadian family. These mutations were not present in a total of 106 normal chromosomes. In another Turkish family with JOAG, we also detected a sequence variant that was proven to be an amino acid polymorphism (Arg76Lys). No other sequence changes were found in the entire coding region and splice junctions of the TIGR/MYOC gene in this family. However, it is still possible that mutations either in the TIGR promoter or in another neighbouring gene could cause glaucoma in this JOAG family. Our results confirm the role of the TIGR/MYOC gene in the aetiology of the JOAG phenotype. PMID- 9863595 TI - 1.4 Mb candidate gene region for X linked dyskeratosis congenita defined by combined haplotype and X chromosome inactivation analysis. AB - Dyskeratosis congenita (DC) is a rare inherited disorder characterised by the early onset of reticulate skin pigmentation, nail dystrophy, and mucosal leucoplakia. In over 80% of cases bone marrow failure develops and this is the main cause of early mortality. The DC1 gene responsible for the X linked form (MIM 305000) of dyskeratosis congenita has been mapped to Xq28. In order to narrow the candidate gene region, genetic linkage analysis was performed in eight X linked pedigrees using a set of markers spanning Xq28. A maximum lod score of 5.31 with no recombinations was achieved with marker DXS1073. Two recombination events were identified; one of these uses X chromosome inactivation pattern analysis to determine carrier status and haplotype analysis to fine map the recombination breakpoint. The fine mapping of these recombination events has enabled the candidate gene region for X linked dyskeratosis congenita to be defined as the 1.4 Mb interval between Xq3274 and DXS1108. PMID- 9863597 TI - Segregation of mutations in arylsulphatase E and correlation with the clinical presentation of chondrodysplasia punctata. AB - Sixteen males and two females with symmetrical (mild) type of chondrodysplasia punctata were tested for mutations in the X chromosome located arylsulphatase D and E genes. We identified one nonsense and two missense mutations in the arylsulphatase E gene in three males. No mutations were detected in the arylsulphatase D gene. Family studies showed segregation of the mutant genes establishing X linked inheritance for these families. Asymptomatic females and males were found in these studies. The clinical presentation varies not only between unrelated affected males, but also between affected males within the same family. We also conclude that clinical diagnosis of chondrodysplasia punctata in adults can be difficult. Finally, our results indicate that brachytelephalangy is not necessarily a feature of X linked symmetrical chondrodysplasia punctata. PMID- 9863596 TI - Linkage analysis in Rett syndrome families suggests that there may be a critical region at Xq28. AB - A whole X chromosome study of families in which Rett syndrome had been diagnosed in more than one member indicated that the region between Xq27 and Xqter was the most likely region to harbour a gene which may be involved in the aetiology of the disease. Further, more detailed studies of Xq28 detected weak linkage and a higher than expected sharing of maternally inherited alleles. It is suggested that there may be more than one gene involved in the aetiology of this syndrome, particularly as the very rare families in which more than one girl is affected often show variable clinical symptoms. PMID- 9863598 TI - The C677T mutation of the 5,10-methylenetetrahydrofolate reductase gene is a moderate risk factor for spina bifida in Italy. AB - OBJECTIVE: To estimate the risk for spina bifida associated with the common mutation C677T of the MTHFR gene in a country with a relatively low prevalence of NTDs. DESIGN: Case-control study. SUBJECTS: CASES: 203 living patients affected with spina bifida (173 myelomeningocele and 30 lipomeningocele); controls: 583 subjects (306 young adults and 277 unselected newborns) from northern and central southern Italy. SETTING: CASES: three spina bifida centres; young adult controls: DNA banks; newborn controls: regional neonatal screening centres. MAIN OUTCOME MEASURES: Prevalence of the C677T genotypes in cases and controls by place of birth; odds ratios for spina bifida and estimated attributable fraction. RESULTS: The prevalence of T/T, T/C, and C/C genotype was 16.6%, 53.7%, and 29.7% in controls and 25.6%, 43.8%, and 30.6% in cases, respectively. We found no differences between type of defect or place of birth. The odds ratio for spina bifida associated with the T/T genotype v C/C plus T/C was 1.73 (95% CI 1.15, 2.59) and the corresponding attributable fraction was 10.8%. No increased risk was found for heterozygous patients (OR=0.79, 95% CI 0.53-1.18). CONCLUSION: This study, as well as the meta-analysis we updated, shows that homozygosity for the MTHFR C677T mutation is a moderate risk factor in Europe, and even in Italy where there is a relatively low prevalence of spina bifida. The estimated attributable fraction associated with this risk factor explains only a small proportion of cases preventable by periconceptional folic acid supplementation. Thus, other genes involved in folate-homocysteine metabolism, their interaction, and the interaction between genetic and environmental factors should be investigated further. PMID- 9863600 TI - Knowledge, views, and experience of 25 women with myotonic dystrophy. AB - Twenty-five affected women of reproductive age known to the North West Regional Genetics Family Register (NWRGFR) were interviewed. A semistructured questionnaire, completed by the interviewer, was used to assess understanding and experience of the clinical and genetic aspects of myotonic dystrophy (MD) and attitudes to prenatal diagnosis (PND). Characteristic features of MD (muscle weakness and wasting and myotonia) were well known. Knowledge of other features and complications reflected experience. All subjects were aware that MD is inherited, but only 56% (14/25) knew the risk to their own children and subjects tended to overestimate this risk. Anticipation and maternal transmission of congenital myotonic dystrophy (CMD) were often misunderstood. Almost half of the subjects (12/25) perceived themselves to be moderately or severely affected and 40% (10/25) felt that their symptoms restricted daily life. Feelings of devastation, depression, worry about the future, and guilt at the risk of transmission to their children were described. Many subjects (10/25) said that the worst aspect of MD is the risk of transmission to their children. Over half (14/25) said that the risk of transmitting MD had influenced or would influence their own reproduction. Three-quarters of subjects who felt that MD had influenced their reproductive decisions (9/12) chose to limit their family or have no children; only 25% (3/12) requested PND. Subjects felt that the lack of information concerning clinical severity made PND for MD difficult to consider. PMID- 9863599 TI - Uniparental disomy for chromosome 6 results in steroid 21-hydroxylase deficiency: evidence of different genetic mechanisms involved in the production of the disease. AB - Congenital adrenal hyperplasia (CAH) is an inherited recessive disorder of adrenal steroidogenesis caused by mutations in the steroid 21-hydroxylase gene (CYP21) in more than 90% of affected patients. The CYP21 gene is located within the HLA complex locus on chromosome 6 (6p21.3). During a molecular characterisation study of a group of 47 Mexican families with 21-hydroxylase deficiency, we identified nine in which the mutation or mutations found in the patient did not appear to originate from one of the parents. Through DNA fingerprinting, paternity was established in all nine families with a probability of non-paternity in the range of 10(-19) to 10(-23). Among these families, we identified one patient with exclusive paternal inheritance of all eight markers tested on chromosome 6p, despite normal maternal and paternal contributions for eight additional markers on three different chromosomes. We did not identify duplication of paternal information for markers in the 6q region, consistent with lack of expression of transient neonatal diabetes owing to genomic imprinting in this patient. Our results substantiate evidence for the existence of different genetic mechanisms involved in the expression of this recessive condition in a substantial portion (approximately 19%) of affected Mexican families. In addition to the identification of a patient with paternal uniparental disomy, the occurrence of germline mutations may explain the unusual pattern of segregation in the majority of the remaining eight families. PMID- 9863601 TI - A family with mental retardation, variable macrocephaly and macro-orchidism, and linkage to Xq12-q21. AB - A family with X linked inheritance of mental retardation (XLMR) is presented. There are 10 mentally retarded males and two affected females in two generations. There are four obligatory carriers, one of whom is described as "slow". Most affected males show macrocephaly and macro-orchidism, which are typical signs of the fragile X syndrome, but have been tested cytogenetically and by analysis of the FMR1 gene and do not have this syndrome. However, some normal males in the family also exhibit macro-orchidism and macrocephaly. Linkage analysis using markers derived from the X chromosome indicates that the causative gene in this family is located in the proximal long arm of the X chromosome, in the interval Xp11-q21. Maximum lod scores of 2.96 with no recombination were found at three loci in Xq13-q21: DXS1111, DXS566, and DXS986. Recombination was observed with DXS1002 (Xq21.31) and DXS991 (Xp11.2), loci separated by about 30 Mb. Although isolation of the gene in this family will be difficult because of the size of the region involved, the localisation should be helpful in investigating other similar families with XLMR, macrocephaly, and macro-orchidism not attributable to FMR1. PMID- 9863602 TI - Agenesis of the corpus callosum with Probst bundles owing to haploinsufficiency for a gene in an 8 cM region of 6q25. AB - Agenesis of the corpus callosum (ACC) is a relatively common brain abnormality resulting from developmental defects either limited to the structures leading to the proper formation of the corpus callosum or involving the embryo forebrain more generally. ACC is genetically heterogeneous with autosomal dominant, autosomal recessive, and X linked inheritance and has also been reported in subjects with aneuploidies involving several chromosomes. Among them, distal 6q deletions have been consistently reported in association with ACC, suggesting that there is a gene in the deleted region whose haploinsufficiency impairs normal corpus callosum development. We have studied a child with ACC with Probst bundles and a deletion at 6q25 of about 8 cM, from D6S1496 to D6S437. Probst bundles are the axons that should have formed the corpus callosum but, unable to cross the midline owing to absence of the massa commissuralis, they run longitudinally along the medial walls of the lateral ventricles from the frontal to the occipital lobes. Thus, their presence suggests that a gene located in the 6q deleted region is specifically involved in the formation of the massa commissuralis and that its haploinsufficiency leads to primary ACC. PMID- 9863603 TI - The butyrylcholinesterase K variant and susceptibility to Alzheimer's disease. AB - Previous work has shown an association between the K variant of the butyrylcholinesterase (BCHE) gene and Alzheimer's disease (AD) in patients carrying the epsilon4 allele of ApoE. We attempted to replicate this finding in 181 UK white AD cases and 71 controls. No difference was found in BCHE-K genotypes (p=0.75) or alleles (p=0.70) between patients and controls. Moreover, despite a significant excess of ApoE epsilon4 in patients versus controls (p<0.0001), we found no evidence to support previous reports of an interaction between ApoE and BCHE-K (chi2=1.49, df=4, p=0.83). PMID- 9863604 TI - Costello syndrome: two cases with embryonal rhabdomyosarcoma. AB - Costello syndrome is a well delineated mental retardation syndrome of unknown aetiology in which the occurrence of benign tumours, especially papillomata, is recognised. We report two children in whom the diagnosis of Costello syndrome was made in the first months of life, who both developed a retroperitoneal embryonal rhabdomyosarcoma. Although not previously reported, the occurrence of this relatively uncommon childhood tumour in two girls with Costello syndrome suggests that an increased risk of malignancy may be part of this condition. The genetic basis of this susceptibility requires further clarification. PMID- 9863605 TI - Familial occurrence of congenital incomplete prepyloric mucosal diaphragm. AB - Incomplete prepyloric mucosal diaphragm (IPMD) is an uncommon congenital anomaly that leads to gastric outlet obstruction in infancy and childhood. This report describes the occurrence of IPMD in six children in a closely knit tribal family from a geographically isolated desert town with a small population in the Sahara. Their records showed similarities of clinical, radiological, operative, and histopathological features. These features, as well as its occurrence in brothers, sisters, and cousins, suggest that this unusual anomaly is transmitted as an autosomal recessive trait. PMID- 9863606 TI - Autosomal recessive anhidrotic ectodermal dysplasia in a large Moroccan family. AB - We studied a large Moroccan family in which anhidrotic ectodermal dysplasia is transmitted as an autosomal recessive trait. Fourteen family members, both males and females, were affected and they all had a common ancestor. Linkage analysis by homozygosity mapping in this family will permit the gene localisation of this rare form of anhidrotic ectodermal dysplasia. PMID- 9863607 TI - Segregation distortion in myotonic dystrophy. AB - Myotonic dystrophy (DM) is an autosomal dominant disease which, in the typical pedigree, shows a three generation anticipation cascade. This results in infertility and congenital myotonic dystrophy (CDM) with the disappearance of DM in that pedigree. The concept of segregation distortion, where there is preferential transmission of the larger allele at the DM locus, has been put forward to explain partially the maintenance of DM in the population. In a survey of DM in Northern Ireland, 59 pedigrees were ascertained. Sibships where the status of all the members had been identified were examined to determine the transmission of the DM expansion from affected parents to their offspring. Where the transmitting parent was male, 58.3% of the offspring were affected, and in the case of a female transmitting parent, 68.7% were affected. Studies on meiotic drive in DM have shown increased transmission of the larger allele at the DM locus in non-DM heterozygotes for CTGn. This study provides further evidence that the DM expansion tends to be transmitted preferentially. PMID- 9863608 TI - Mild phenotype associated with an interstitial deletion of the long arm of chromosome 1. AB - We report on a 21 month old child referred to us because of facial dysmorphism and psychomotor retardation. The patient's phenotype was characterised by a wide and receding forehead, broad nasal bridge, redundant retronuchal skin, low set and poorly shaped ears, micrognathia, and small hands and feet. High resolution R and G banding karyotype analysis of peripheral blood lymphocytes showed an interstitial deletion of the long arm of chromosome 1 spanning bands q22 to q24. The cytogenetic results were confirmed by molecular analysis. The phenotype observed in our patient was relatively milder than those reported in other patients with an interstitial deletion of chromosome 1q. PMID- 9863609 TI - De novo unbalanced translocation resulting in monosomy for proximal 14q and distal 4p in a fetus with intrauterine growth retardation, Wolf-Hirschhorn syndrome, hypertrophic cardiomyopathy, and partial hemihypoplasia. AB - We present the perinatal findings of a fetus with a de novo unbalanced chromosome translocation that resulted in monosomy for proximal 14q and monosomy for distal 4p. Prenatal sonographic examination at 27 weeks of gestation showed intrauterine growth retardation, microcephaly, cardiomegaly with arrhythmia, and asymmetry of the upper limbs. Genetic amniocentesis showed an abnormal karyotype of 45,XX,der(4)t(4;14)(p16.3;q12),-14. Linkage analysis of the family confirmed the maternal origin of the deletions. Molecular refinement of the deletion breakpoints indicated that the breakpoints at 4p16.3 and 14q12 were located between loci D4S403 (present) and D4S394 (absent), and between loci D14S252 (present) and D14S64 (absent), respectively. Necropsy showed dysmorphic features compatible with Wolf-Hirschhorn syndrome, hypertrophic cardiomyopathy, partial hemihypoplasia, and a normal brain without evidence of holoprosencephaly. Our case adds to the list of clinical phenotypes associated with the proximal regions of 14q. PMID- 9863610 TI - Kenny-Caffey syndrome without the CATCH 22 deletion. PMID- 9863611 TI - Kenny-Caffey syndrome is part of the CATCH 22 haploinsufficiency cluster. PMID- 9863612 TI - Kenny-Caffey syndrome is part of the CATCH 22 haploinsufficiency cluster. PMID- 9863613 TI - Genotypic/phenotypic heterogeneity of Kenny-Caffey syndrome. PMID- 9863614 TI - Tricuspid atresia in sibs. PMID- 9863615 TI - Collagen disease: the enemy within. AB - Surprisingly, the autoimmune diseases predominate in women of childbearing years. Recent evidence suggests that these diseases are the result of some interaction of the hypothalamic-gonadal axis with the immune system. The median age for rheumatoid arthritis is 45 years, the median age for lupus erythematosus is 25. Other illnesses, which are autoimmune in character, such as Sjogren syndrome, scleroderma and the vasculitides, are also more commonly found in women. There is no link that ties these illnesses together, except for gender and various disparate immune manifestations such as autoantibodies. The etiopathogenesis of these diseases is reviewed. These diseases are notoriously difficult to diagnose; they mimic other illnesses in their early presentations. Accompanying illnesses such as migraine headaches, Hashimoto's thyroiditis, and fibromyalgia are discussed as related entities. Immunosuppression of diseases like rheumatoid arthritis and lupus erythematosus, is discussed. Various methods of management are considered, such as the use of steroids, cytotoxic agents, and new experimental agents, such as DHEA and IVIG. PMID- 9863616 TI - Effect of in vitro incubation on spontaneous acrosome reaction in fresh and cryopreserved human spermatozoa. AB - OBJECTIVES: Capacitation and acrosome reaction are prerequisites for fertilization. However, in vitro capacitation is not necessary for an agonist induced acrosome reaction. We studied whether in vitro capacitation is important in spontaneous acrosome reaction and analyzed how capacitation before cryopreservation influences the acrosomal status of thawed spermatozoa. METHODS: Semen specimens from normal donors (n = 15) were processed by the swim-up technique and divided into two aliquots. One aliquot was capacitated (capacitation induced) for three hours by incubation in a modified-BWW medium with 3% HSA at 37 degrees C under 5% carbon dioxide. The other aliquot did not receive any treatment. Both aliquots were analyzed by CASA to assess the capacitation status of the spermatozoa and then cryopreserved. Spontaneous acrosome reaction was assessed by FITC-PNA lectin before and after cryopreservation. Sperm viability was measured using Hoechst-33258 stain. RESULTS: Before freezing, the frequency of spontaneous acrosome reaction was higher in the capacitation-induced sperm preparation (median, 20.5% [interquartile range, 17.2-37.8]) than in swim-up-induced specimens (median, 10.6% [range, 4.8-23.2]; P <.001). The percentage of viable cells showing acrosome reaction increased after cryopreservation in both swim-up-induced specimens (median, 241.4% [interquartile range, 37.1-678.6]; P <.001) and capacitation-induced specimens (median, 48.2% [range, 6.1-63.3]; P = 0.002). Although this increase was higher in the swim-up-induced specimens (P = 0.002), frequency of postthaw spontaneous acrosome reaction was similar in both groups (P = 0.18). CONCLUSIONS: We conclude that sperm capacitation significantly optimizes the acrosome reaction. However, a small proportion of normal spermatozoa do not require capacitation to undergo spontaneous acrosome reaction in vitro. After cryopreservation, the percentage of spermatozoa that had intact acrosomes was similar in both groups, despite the fact that one aliquot underwent prefreeze capacitation. These findings suggest that the acrosome reaction may involve complex mechanism(s) rather than a physiological change induced by capacitation. PMID- 9863617 TI - Antiviral therapy for genital herpes in nonpregnant and pregnant women. AB - There have been major developments in basic and clinical research in the field of antiviral therapy. The health care provider needs to know the availability of numerous antiviral compounds and begin to view them as we do antibiotics. They need to be used selectively to modify infections and prevent the morbidity and mortality associated with viral infections, such as herpes simplex virus (HSV, genital herpes). Therapy of these infections is required to treat not only the patient but also to prevent infection of sexual partners and the newborn. Numerous and varied antiviral compounds have been approved by the FDA to treat genital HSV infections. They all have the same mechanism of action. Acyclovir, valacyclovir, and famciclovir have been approved for the treatment of primary and recurrent disease. All three agents can be used for daily administration for the suppression of recurrent disease. Use of these antiviral compounds in the nonpregnant and pregnant woman can significantly alter the disease and influence transmission rates along with decreasing the morbidity and mortality associated with HSV infections. PMID- 9863618 TI - Achieving and maintaining pregnancy after age 40. AB - Demographic data from the United States show the need for testing ovarian reserve in a population that is increasingly delaying reproduction. The childless rate for U.S. women over the age of 35 has nearly doubled to 25% since 1925. Yet, in 1994 approximately one-third of all first births were by women over the age of 35. The first-line treatment for the aging effect on the ovarian reservoir of oocytes is ovulation induction. Study results reveal that a delivery rate of only 2.4% was achieved at age 42, and in 119 cycles of therapy in women above that age there were no live births. The clomiphene citrate challenge test is one method of dynamic testing of ovarian reserve. It has the advantages of simplicity and reduced cost when compared with the gonadotropic stimulatory test. In one test group utilizing the clomiphene citrate challenge test only 4.8% conceived at any age following an abnormal test. As a prognostic test with an approximate 95% accuracy rate for negative predictive value it has much to offer. With oocyte donation becoming more widespread as an acceptable method of achieving pregnancy, clomiphene citrate challenge testing should be performed more frequently, in order to spare couples the emotional and financial sequelae of multiple attempts at ovulation induction and IVF when expected pregnancy rates fall below a level acceptable to both patient and physician. PMID- 9863619 TI - The evaluation of the sperm migration test as a predictor for success with intrauterine insemination. AB - OBJECTIVE: The objective of this study was to prospectively evaluate the sperm migration test (SMT) as a discriminator in couples undergoing intrauterine insemination (IUI). PATIENTS AND METHODS: 261 couples underwent 797 IUI treatment cycles involving gonadotropin stimulation in the three year period. All had a diagnosis of unexplained infertility. All male partners underwent a repeat standard seminal analysis and SMT prior to the female partner undergoing controlled ovarian stimulation. RESULTS: Despite apparently normal seminal analyses before referral, in 22 samples the sperm concentration, motility or morphology were abnormal (WHO criteria). Of these, 20 couples underwent 109 cycles and achieved 2 pregnancies giving a pregnancy rate of 1.8% per cycle and a cumulative pregnancy rate of 10% per couple. From the remaining couples with normal seminal analyses, 71 had an SMT <5 million/mL and 168 had an SMT >5 million/mL. The suboptimal SMT group underwent 276 cycles (3.89 cycles per couple) and achieved 18 pregnancies giving a pregnancy rate of 6.5% per cycle and a cumulative pregnancy rate of 25.4%. The normal SMT group underwent 412 cycles (2.45 cycles per couple) and achieved 60 pregnancies giving a pregnancy rate of 14.6% per cycle and a cumulative pregnancy rate of 35.7%. CONCLUSIONS: We confirm that abnormal seminal analysis leads to poor pregnancy rates with IUI. However, an SMT <5 million/mL despite normal seminal analysis (WHO criteria) also leads to significantly worse pregnancy rates. We would recommend that prior to IUI, couples are screened using the SMT. PMID- 9863620 TI - Immunologic variables in normal pregnancy and spontaneous abortion. AB - OBJECTIVE: The aim of the study was to establish if pregnancy belongs to pathological situations, if it changes SIL-2R, sCD4 and sCD8 levels, and to verify if these levels have a prognostic value in the evaluation of pregnancy. PATIENTS AND METHODS: Thirty pregnant women in the first trimester of pregnancy were studied. Ten of them had a normal evolution of pregnancy, ten had a threatened abortion with a bad outcome, and ten had an ectopic pregnancy. We determined SIL-2R, sCD4, and sCD8 levels in the serum and in the amniotic fluid in 10 pregnant women in their second trimester, and in 10 healthy women without pregnancy (control group). RESULTS: We found that (a) 50% of the pregnant women in their first trimester had abnormal SIL-2R values, and 90% had abnormal sCD8 values; (b) 9/10 women with threatened abortion and bad outcome had abnormal SIL 2R values; (c) SIL-2R levels were significantly higher in the amniotic fluid than in the serum (P <.001). CONCLUSION: Our results sustain the opinion that pregnancy must be included among those situations that increase SIL-2R, sCD4, and sCD8 levels. SIL-2R levels greater than 1,300 U/mL carried a bad prognosis in the evolution of pregnancy. PMID- 9863621 TI - Correlation between peritoneal fluid and serum antiphospholipid antibodies in women with primary infertility. AB - OBJECTIVE: To compare the peritoneal and serum levels of antiphospholipid antibodies (aPLs) against the following phospholipids: cardiolipin, L-alpha phosphatidic acid, L-alpha-phosphatidylethanolamine, 1-alpha-phosphatidyl-DL glycerol, L-alpha-phosphatidyl-inositol, L-alpha-phosphatidyl-L-serine in immunoglobulin isotypes of G, M, and secretory A. PATIENTS AND METHODS: A group of 107 women with primary infertility were immunologically examined. An ELISA method was used for detection of aPLs. RESULTS: aPL-L-serine and aPL-cardiolipin predominate over peritoneal fluid in all studied Ig isotypes (G, M, secretory A). Levels of aPL-ethanolamine are much higher in peritoneal sIgA and IgM than in sera. There is a predominance of IgG-aPLs-inositol, DL-glycerol, and aPL-acid in serum, and of sIgA in peritoneal fluid. CONCLUSION: Our patients had higher levels of aPLs (IgG, IgM, sIgA) against cardiolipin, serine, and ethanolamine in peritoneal fluid than in sera. The prevalence of aPLs in IgG was against DL glycerol, phosphatidyl-inositol, and phosphatidic acid. It is quite difficult to estimate whether the aPLs have an influence directly through phospholipid epitopes on uterine mucous membrane, on the surface of oocytes, or on the surface of the early embryo during the fertilization and/or implantation process. PMID- 9863622 TI - Mechanisms mediating multiple physiological responses to gonadotropin-releasing hormone. AB - A central question in endocrinology is how a single ligand interacting with a single receptor can mediate multiple responses. GnRH interaction with receptor offers a prime example, leading to the regulation of synthesis and release of at least three molecules, regulation of target cell responsiveness and receptor number. The present study suggests a molecular model consistent with extant data that provides a mechanism by which this may occur and, further, which allows for coordinate regulation. PMID- 9863623 TI - Functional analysis of GnRH receptor ligand binding using biotinylated GnRH derivatives. AB - The objective of this study was to determine whether the gonadotrophin-releasing hormone (GnRH) ligand binds to the GnRH receptor (GnRH-R) with either the N- and C-termini or the beta-II turn pointing towards the cell. The functionality of GnRH and two biotinylated GnRH derivatives, biotin [D-Lys6]GnRH and biotin [Gln1]GnRH biotinylated at positions 6 and 1, respectively was assessed. Streptavidin was also used in combination with these peptides to investigate the effects of the steric hindrance caused by this molecule on ligand binding when bound to the biotin molecules at the two positions. GnRH bound to the receptor with high affinity, which was not affected by the addition of streptavidin. Both the biotinylated derivatives bound to the receptor though with lower affinities than GnRH. The biotin [D-Lys6]GnRH-streptavidin complex bound to the receptor albeit with lower affinity compared to biotin [D-Lys6]GnRH only, although it maintained its ability to cause receptor internalisation. The ability of the biotin [Gln1]GnRH to bind to the receptor was abolished in the presence of excess streptavidin. Both GnRH and biotin [D-Lys6]GnRH stimulated total inositol phosphate production whereas biotin [Gln1]GnRH exhibited GnRH antagonist activity. It appears that the small biotin molecule can be accommodated within the binding pore when attached to position 1 of the ligand but not when complexed to streptavidin. The fact that biotin [D-Lys6]GnRH maintains functionality when complexed to streptavidin while biotin [Gln1]GnRH does not, suggests that the N- and possibly the C-termini are required for receptor binding. Thus the most likely binding orientation for the ligand is with the N- and C-termini pointing inwards with the residue at position 6 pointing away from the binding site. PMID- 9863624 TI - Structure and function of the ovine type 1 corticotropin releasing factor receptor (CRF1) and a carboxyl-terminal variant. AB - Corticotropin releasing factor (CRF) is the major neuropeptide regulating the hypothalamo-pituitary-adrenocortical axis in most species. A pituitary receptor for CRF (designated CRF1) belonging to the seven-transmembrane helix, G-protein coupled receptor superfamily has been cloned for human, rat, mouse and xenopus. Since ovine CRF shares only 84% identity to human/rat CRF (h/rCRF) we postulated that the sheep pituitary CRF1 receptor may have similarly diverged from the rodent and human CRF1. We report the molecular cloning of an ovine pituitary cDNA containing a 1245 bp open reading frame encoding a 415 amino acid sheep CRF1 receptor 78, 86, 94, and 95% homologous to xenopus, chicken, rat, mouse, and human CRF1, respectively. The divergence in primary structure between the sheep CRF1 and the other mammalian CRF1s is primarily localized to the extracellular amino terminal domain of the receptor (18 of 22 divergent residues, ovine vs human CRF1). A variant of the oCRF1 was also isolated (oCRF1var) with 133 bp deleted from nucleotide (nt) 1080 to nt 1213 of the open reading frame (ORF) resulting in a new ORF of 1176 nt predicting a 392 residue CRF1 variant receptor. The 133 bp deletion would cause a frame-shift at residue 358 within the carboxyl third of the seventh transmembrane domain (TM7) resulting in a shortened cytoplasmic tail with a new amino acid sequence from residue 358 to 392. Scatchard analysis of saturation curves using membrane prepared from Cos 7 cells transfected with oCRF1 or oCRF1var indicated that both wild-type and variant receptors were expressed similarly (number of CRF binding sites) and both bound oCRF with high affinity [oCRF1 (Kd): 2.5 + 1.6 nM; oCRF1var: 5.1 + 2.3 nM]. The non-hydrolyzable GTP analogue (GTPgammaS) lowered the affinity of both wild-type and variant oCRF1 receptors to a similar extent (oCRF1: 18.2 nM; oCRF1var: 22.4 nM). Both wild-type and variant oCRF1 receptors exhibited approximately 10-fold greater selectivity for oCRF and sauvagine compared to h/rCRF or alpha-helical [9 41]oCRF. CRF effectively stimulated the accumulation of cAMP (EC50 = 51 pM) in Cos 7 cells transiently transfected with wild-type but not variant oCRF1 receptor. In Cos 7 cells transfected with oCRF1var, cAMP accumulation was only observed at the highest concentration of oCRF utilized (100 nM). Basal (unstimulated) levels of cAMP in Cos 7 cells transfected with oCRF1var (in the presence of 2 mM IBMX) were approximately 50% lower than for the wild-type oCRF1. Differences in cAMP accumulation could not be attributed to differences in receptor number since total binding sites in the transfected cells were not different between wild-type or variant oCRF1 receptors. Agonist-induced receptor internalization, determined as the percent of total [125I] Tyr0-oCRF bound located in the acid-resistant fraction of transfected Cos 7 cells, increased with time (0-60 min at 37 degrees C) for both wild-type and variant oCRF1. Wild-type CRF1 internalized approximately 2-fold greater percent of total [125I] Tyr0-oCRF bound compared to the variant receptor. In summary, an ovine CRF1 and a CRF1 cytoplasmic tail receptor variant displaying high affinity binding to oCRF as well as selectivity for oCRF vs h/rCRF, were cloned from an adult sheep pituitary cDNA library. GTPgammaS studies indicate that both variant and wild-type receptors couple efficiently to Galphas however, only the wild-type oCRF1 is capable of stimulating cAMP production at physiological levels of CRF. Agonist induced internalization of the ovine CRF1var is also reduced compared to the wild type CRF1 receptor. We suggest that the oCRF1var interacts efficiently with Galphas but is unable (post-hormonal binding) to effectively stimulate G-protein activation of adenylate cyclase, indicating that the cytoplasmic tail of the CRF1 can modulate receptor function related to signal transduction. (ABSTRACT TRUNCATED) PMID- 9863625 TI - Adrenal angiotensin II type 1 and type 2 receptors in Cushing's and Conn's syndromes. AB - Cushing's and Conn's syndromes are well recognised endocrine diseases, but the pathobiology of the tumors causing these disorders is unclear. In this study we examined AT1 and AT2 gene expression in adrenal adenomas of Cushing's and Conn's syndromes. AT1 and AT2 receptor mRNA, as well as alternatively spliced AT1 transcripts, were detected by RT-PCR using adjacent adrenal cortex tissue as controls. Whereas no consistent differences in AT1 mRNA were seen compared to control adrenal cortex, AT2 mRNA levels were significantly decreased in the adenomas of Cushing's and Conn's syndromes. No changes in alternative splicing of AT1 mRNA were observed in the adrenal tumors. The fact that no consistent changes were seen in AT1 mRNA or its splicing, whereas AT2 mRNA were reduced in both forms of hormone producing adrenal tumor suggests that the AT2 receptor, rather than the AT1 subtype, may be correlated with adrenal tumorigenesis. PMID- 9863626 TI - Neurotensin enhances agonist-induced cAMP accumulation in PC3 cells via Ca2+ dependent adenylyl cyclase(s). AB - A human prostate cancer cell line (PC3) with abundant neurotensin (NT) receptors was used to demonstrate that NT potentiated 3',5'-cyclic adenosine monophate (cAMP) accumulation in response to a variety of stimuli, including both direct forskolin (F) and indirect (prostaglandin, (PGE2), isoproterenol (ISO) and cholera toxin (CTx)) activators of adenylyl cyclase. Several mechanisms were investigated and our results indicated an effect on the rate of cAMP formation and not on degradation or extrusion. For each stimulus, NT enhanced efficacy without altering EC50. The effect of NT did not involve stimulatory G-protein (Gs)-activation or interference with a tonic inhibitory G-protein (Gi)-mediated inhibition. A similar response was obtained when NT was added with the stimulus or given as a two minute pulse which was removed prior to addition of stimulus. The potentiating activity disappeared with a t1,2 of approximately 15 min. NT transiently elevated cellular [Ca2+]i and its effects on cAMP could be mimicked by [Ca2+]i-elevating agents (uridine triphosphate (UTP), thapsigargin and ionomycin). Buffering cellular [Ca2+]i with 1,2-bis (2-aminophenoxy) ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM) inhibited cAMP responses to ISO and F in presence and absence of NT. These data support the idea that NT potentiated cAMP formation in response to a variety of stimuli by facilitating the activation of Ca2+ -dependent adenylyl cyclases. PMID- 9863627 TI - Time-course effects of human recombinant luteinizing hormone on porcine Leydig cell specific differentiated functions. AB - Since recombinant hormones are considered as safer and more reliable in their bioactivity than extractive hormones, the recently available human recombinant luteinizing hormone (r-hLH), will probably replace hCG in the near future, for clinical purposes. This prompted us to investigate whether or not, and by which mechanisms, r-hLH can induce a desensitization of signal transduction and/or an up-regulation of steroidogenic capacity in Leydig cells. The effects of a 30 min to 24 h exposure to r-hLH (10(-9) M) on the differentiated functions of cultured immature porcine Leydig cells were studied by measuring the following parameters: LH/hCG receptor number and mRNA, hCG-, cholera toxin- and forskolin-induced cAMP production, G protein alphas subunit content of the membrane, hCG-, cholera toxin , forskolin-, 8Br-cAMP-, 22R-OH-cholesterol-, progesterone-, 170H-progesterone-, DHEA-, delta4-androstenedione-induced testosterone secretion and StAR, 3beta-HSD, cytochrome P-450scc and P-450c17 mRNAs. hCG binding sites and LH/hCG receptor mRNA were slowly down regulated by r-hLH, reaching 47+/-1 and 18+/-7% of control at 24 h, respectively. Down-regulation of both hCG- and cholera toxin-induced cAMP production occurred earlier and was more marked, and at 24 h represented only 2.7+/-0.5 and 12.5+/-3.6% of control. Due to the synergistic effect of r-hLH and forskolin on cAMP production, the forskolin-induced cAMP was higher in r-hLH treated than in control cells, but this response also declines with time and was, at 24 h, only 32% of that observed at 30 min. This decreased cAMP production was associated with a less marked decline in the amount of membrane content of Galphas protein. The testosterone production in response to hCG, cholera toxin, forskolin and 8Br-cAMP declined to reach a nadir at 6 h but increased thereafter and at 24 h was significantly higher than in control cells. In contrast, the conversion of several precursors into testosterone remained stable or increased slightly during the first hours of r-hLH treatment and significantly increased at 24 h and this was associated with an increase of StAR, 3beta-HSD, P-450scc and P 450c17 mRNAs. Taken together, the present results indicate that, despite the marked down-regulation of transmembrane signaling, r-hLH increased the steroidogenic capacity of Leydig cells by increasing the expression of several genes encoding the proteins involved in testosterone synthesis. PMID- 9863628 TI - Tissue-specific modulation of rat glucagon receptor mRNA by thyroid status. AB - The influence of thyroid status on glucagon receptor mRNA levels was investigated in rats using a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Glucagon receptor mRNA was detected in liver, brown and white adipose tissues (BAT and WAT) and brain. In BAT and WAT, pharmacologically induced moderate hypothyroidism resulted in a marked reduction in the relative abundance of glucagon receptor mRNA. Short-term treatment of hypothyroid rats with exogenous 3,3',5'-triiodo-L-thyronine (T3), resulting in a marked hyperthyroidism, reversed the phenomenon in BAT while the reversal was only partial in WAT. In the liver, there was no significant effect of mild hypothyroidism while there was a positive effect of hyperthyroidism. In brain, the relative tissue abundance of glucagon receptor mRNA was not affected by the large changes in plasma T3. The present results therefore indicate that thyroid status may modulate the relative abundance of glucagon receptor mRNA in a tissue specific manner. PMID- 9863629 TI - Genistein exerts estrogen-like effects in male mouse reproductive tract. AB - The aim of this study was to evaluate the estrogenicity of genistein in the neonatal and adult male mouse reproductive tract. In intact adults, genistein (2.5 mg s.c./kg of body weight/day for 9 days) reduced testicular and serum testosterone concentrations, pituitary LH-content and prostate weight. In castrated adults, genistein (0.025-2.5 mg s.c./kg of body weight) increased expression of c-fos gene in prostatic urethra. In adult, neonatally estrogenized mice showing an increased estrogen sensitivity, a 10-day treatment with genistein (2.5 mg s.c./kg of body weight) induced development of squamous epithelial metaplasia in prostatic collecting ducts. Neonatally, only a very high dose of genistein (1 mg/pup per day; i.e. approximately 500 mg/kg of body weight) induced persistent structural changes, similar to those seen in mice treated neonatally with diethylstilbestrol, in the urethroprostatic complex. These results suggest that in adult males, genistein induces the typical estrogenic effects in doses comparable to those present in soy-based diets, while in neonatal animals, considerably higher doses are required to show estrogen-like activity. PMID- 9863630 TI - Multiple elements in the distal part of the 1.2 kb 5'-flanking region of the rat GnRH receptor gene regulate gonadotrope-specific expression conferred by proximal domain. AB - Several lines of evidence indicate that the number of GnRH receptors (GnRH-R) and therefore, gonadotrope responsiveness to GnRH, is highly dependent upon the level of GnRH-R mRNA. To explore this aspect of regulation, we have isolated a 3.3 kb fragment encompassing the promoter region of the rat GnRH-R gene. Primer extension and RNase protection assays allowed the identification of five major transcriptional start sites located within the 110 bp region upstream of the translation start codon. Transfection experiments using the CAT reporter gene demonstrated that the 1261 bp 5' flanking region is required to direct high efficient expression in the gonadotrope-derived alphaT3-1 cell line thus contrasting with mouse in which the only 500 bp proximal sequence appeared to be sufficient. Another difference between rat and mouse was apparent in the 183 bp region of the rat promoter which induced a 3-fold stimulation of thymidine kinase promoter activity in both alphaT3-1 and CHO cells. Subsequent deletion analysis of the region residing between -1261 and -519 revealed the presence of multiple regulatory domains that contributed to the cell-specific activity. However, despite this efficiency in the context of the wild-type promoter, they failed to induce the activity of the minimal thymidine kinase (TK) promoter in the absence of the proximal 600 bp promoter region. Accordingly, a composite TK promoter containing the entire 1.2 kb promoter induced a 10-fold increase in the activity of the TK promoter in alphaT3-1 cells. Taken together these data suggest that distal regulatory regions are critical and require cooperation with proximal specific-promoter elements for activating basal R-GnRH gene expression in gonadotrope cells. PMID- 9863631 TI - Effect of chronic GH overproduction on cardiac ANP expression and circulating ANP levels. AB - Sodium and water retention are common in acromegaly and upon GH administration. The underlying mechanisms, however, have not been clearly characterized as yet. Therefore, the aim of this study was to examine possible alterations of atrial natriuretic peptide (ANP), an endogenous regulator of volume homeostasis, in response to chronic elevated GH. We used GH-transgenic mice (GH-TM) as a model for chronic hypersomatotropinemia and moreover investigated 7 and 27 week old animals, respectively, in order to discriminate between short and long term effects of GH overexpression. Hematocrit values were reduced in GH-TM compared to control animals and it is known that plasma volume is increased in these animals. Structural lesions of the kidney were found in the GH-TM, however, in the animals studied there were no signs of renal insufficiency as evidenced by serum creatinine and urea levels. The serum concentration of immunoreactive ANP (IR ANP) determined by RIA was significantly (P < 0.005) elevated in the young GH-TM as compared to control littermates (81.7+/-13.3 vs. 50.9+/-10.8 fmol/ml). The increase in serum IR-ANP of 27 week old GH-TM, however did not reach the level of significance (57.13+/-16.3 vs. 50.25+/-16.4 fmol/ml). Serum samples of control mice as well as of the 7 week old animals mainly contained ANP 99-126, known to be the circulating form of ANP. In contrast, serum of 27 week old GH-TM predominantly showed the cardiac storage form of ANP, ANP 1-126. Cardiac expression of ANP was quantified by Northern blot analysis. mRNA coding for ANP was found 1.2- and 2-fold increased in the atria of 7 and 27 week old GH-TM, respectively. In parallel, a 2.2-fold (7 week) and 2-fold (27 week) increase of IR-ANP was observed in transgenic atria compared to tissue of control animals. In contrast, no significant difference of ANP mRNA expression or of content of IR ANP was observed in the ventricles of both groups of animals. In conclusion, GH TM show various alterations in their ANP status suggesting an influence of the peptide on the effect of GH in fluid retention. PMID- 9863632 TI - Tautomycin inhibits phosphatase-dependent transformation of the rat kidney mineralocorticoid receptor. AB - The binding of aldosterone (ALDO) to the mineralocorticoid receptor (MR) induces a conformational change of the protein referred to as 'transformation'. This feature can be evidenced in vivo by the capacity of the MR to interact with chromatin, and in vitro by the ability of the MR to bind to DNA strands or to shift the sedimentation coefficient (S) to lower values. The transformation process allows MR to work as a transcription factor after interacting with specific sequences of DNA. The signal transduction pathway for the MR transformation remains unknown. As a first step towards elucidating the mechanism of steroid-dependent MR transformation, we asked if the MR-signaling pathway is affected by the phosphorylation status of the MR-heterocomplex, and how that pathway may be regulated. Incubation of preformed [3H]ALDO-MR complex with bovine intestinal alkaline phosphatase led to an increase in the rate of MR transformation (measured as 9.4-5.4S shift). This alkaline phosphatase-dependent MR transformation was inhibited by the specific alkaline phosphatase-type inhibitor levamisole, and was not evident in incubations performed with acid phosphatases. A direct correlation between the DNA-cellulose binding capacity of the [3H]ALDO-MR complex and the percentage of transformed 5.4S MR form was also observed. When rat kidney cytosol was incubated in the absence of both exogenous phosphatase and stabilizing agents (such as molybdate or vanadate), MR transformation also took place, in a time- and temperature-dependent process. In contrast with the inhibitory effect observed upon alkaline phosphatase-promoted transformation, levamisole was unable to inhibit the endogenous transforming activity of MR, suggesting that an endogenous phosphatase other than those which belong to the alkaline-type may be responsible for that transformation. Tautomycin, a polyketide produced by the soil bacteria Streptomyces which inhibits serine/threonine phosphatases of the PP1/PP2A subgroup, was able to inhibit the endogenous phosphatase activity in a concentration-dependent form (Ki(app)=7.35 nM). These results support the idea that the endogenous renal activity involved in the regulation of rat kidney MR transformation may be a protein phosphatase which belongs to the PP1/PP2A subgroup. PMID- 9863633 TI - A fusca gene homologue in mammalian tissues: Developmental regulation in the rat testes. AB - By differential screening of a rat pineal cDNA library we identified earlier a novel transcript having a 57% nucleotide homology and a 45% amino acid identity with a plant fusca-gene (fus6) to which a corresponding human sequence (gps1) has recently been reported. Expression of this mammalian fusca homologue (mfh) was seen in a variety of mammalian tissues, including kidney, pineal and retina, but it was particularly strong in the testes. Northern blot analysis demonstrated that the rat testicular mfh message increases markedly from day 28 onwards. Additionally, by in situ hybridization, mfh was localized primarily to the seminiferous tubules with a stage-dependent distribution pattern, a result which was confirmed by immunohistochemistry with antibodies raised against a synthetic MFH oligopeptide. Western blotting also revealed strong signals of the expected molecular weight in testicular extracts from several species. In view of its homology to fus6, a plant gene known to be involved in repressing photomorphogenesis in darkness, the conservation of mfh in mammals suggests a potential function for MFH in signaling pathways involved in the regulation of mammalian differentiation and development. PMID- 9863634 TI - Longitudinal study of tissue- and subunit-specific obesity-induced regulation of the pyruvate dehydrogenase complex. AB - The tissue-specific expression of the mitochondrial pyruvate dehydrogenase complex (PDHc) has been studied in an animal model of obesity with hyperinsulinemia, the obese (fa/fa) Zucker rat. Liver and heart were obtained from 4 and 8 week-old obese rats and age-matched lean animals, and in each tissue the following parameters were analyzed: (1) total activity of the mitochondrial PDHc; (2) abundance of the mitochondrial PDHc subunits on Western blots; and (3) abundance of the E1alpha and E1beta subunit mRNAs on Northern blots and semi quantitative RT-PCR. Regardless of age, obese rats showed an increase in liver total PDHc activity and a coordinate increase in liver E1alpha and E1beta PDHc subunit abundance. At 4 weeks, obese rats also showed an increase in liver PDH E1alpha mRNA level, but regardless of age E1beta mRNA level was unchanged. In contrast, neither total PDHc activity nor the concentration of its protein subunits were increased in heart of obese rats. Thus, obese Zucker rats display a liver-specific early increase in PDHc which results from a selective up regulation of the E1alpha gene expression. PMID- 9863635 TI - Prepubertal genistein treatment modulates TGF-alpha, EGF and EGF-receptor mRNAs and proteins in the rat mammary gland. AB - We have previously demonstrated that exposure to genistein early in life protects against chemically-induced mammary cancer in rats. To gain insight into the mechanism of action, we have investigated the expression of the EGF-signaling pathway in the mammary glands of 21 and 50 day old rats treated on days 16, 18, and 20 postpartum with 500 microg genistein/g body weight (B.W.) or an equivalent volume of the vehicle, dimethylsulfoxide (DMSO). This prepubertal genistein treatment up-regulated TGF-alpha and the EGF-receptor (EGFR), but not EGF, in mammary terminal ductal structures at day 21 postpartum. TGF-alpha, EGF and EGFR mRNA levels were similar in 21 day old control- and genistein-treated animals. At day 50 postpartum, mammary glands of genistein treated rats had more lobules and fewer terminal end buds (TEBs) and terminal ducts (TDs), i.e. they were more differentiated. TGF-alpha mRNA levels were down-regulated in TEB of proestrous and estrous females; EGF mRNA levels were down-regulated in TDs of proestrous, but not in estrous females; and EGFR mRNA levels were not altered in 50 day old proestrous or estrous female rats. EGFR immunostaining intensity was decreased in TEBs, but not in the total gland. EGF was increased in TEBs and TDs. TGF-alpha, EGF and EGFR were also observed in the stroma and fat pad, but genistein treatment did not alter the expression of these proteins in those locations. TGF alpha, but not EGF and EGFR, immunostaining was observed in cell nuclei (not modulated by genistein), suggesting that this growth factor may act directly on nuclear events such as transcription and DNA replication. For comparative purposes, prepubertal diethylstilbestrol treatment was investigated and found to decrease EGFR immunostaining intensity and total IHC staining in all terminal ductal structures. We conclude that prepubertal genistein treatment directly stimulates TGF-alpha and EGFR to enhance mammary gland differentiation. This programs the differentiated cells for a down-regulated EGF-signaling pathway in TEBs and TDs of adult mammary glands. Reduced EGFR expression at time of carcinogen exposure may account for genistein programming against mammary cancer. PMID- 9863636 TI - The influence of lobeline on nucleus accumbens dopamine and locomotor responses to nicotine in nicotine-pretreated rats. AB - In vivo brain microdialysis was used to investigate the influence of lobeline on dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) overflow in the core of the nucleus accumbens of freely-moving rats pretreated with nicotine (0.4 mg x kg( 1), s.c., once per day for 5 days). Locomotion was also recorded. Lobeline, at doses of 0.7, 4.0 and 10.0 mg x kg(-1), i.p., failed to elicit any significant changes in extracellular dopamine or dihydroxyphenylacetic acid levels during the 60 min following its administration and did not stimulate locomotor. The dopamine responses to nicotine (0.4 mg x kg(-1), s.c.), were abolished (P<0.01) if the nicotine challenge was administered 10 min but not 60 min, after lobeline doses of 4.0 and 10.0 mg kg(-1), i.p., but were unaffected following lobeline at the lowest dose tested (0.7 mg x kg(-1), i.p.) at either time. The increase in locomotor activity was significantly attenuated (P<0.01), to a similar extent, when the nicotine was injected 10 min, but not 60 min, after all three doses of lobeline (0.7, 4.0 and 10.0 mg kg(-1), i.p.) when compared with the saline treated rats. The results suggest that lobeline is a short-acting antagonist of the nicotinic AChRs which mediate the effects of nicotine on mesolimbic dopamine activity and locomotor stimulation. PMID- 9863637 TI - Lack of effect of a selective vasopressin V1A receptor antagonist SR 49,059, on potentiation by vasopressin of adrenoceptor-mediated pressor responses in the rat mesenteric arterial bed. AB - The vasopressin receptor subtype involved in the enhancement by vasopressin of adrenoceptor-mediated vasoconstriction was investigated in rat isolated perfused mesenteric arteries. [Arg8]vasopressin (1-10 nM) dose-dependently increased the perfusion pressure and enhanced the pressor response to the adrenoceptor agonist methoxamine (40 nmol) or electrical stimulation of periarterial nerves (16 Hz), at the concentration of 10 nM of [Arg8]vasopressin up to 4 and 3 fold, respectively. During prolonged exposure (45 min) the direct vasoconstrictor effect of [Arg8]vasopressin (10 nM) rapidly declined whereas the potentiation of methoxamine-induced vasoconstriction was maintained. The selective vasopressin V1A receptor antagonist SR 49,059 (1-3 nM) and the non-selective V1A/B and oxytocin receptor antagonist [deamino-Pen1,Tyr(Me)2,Arg8]vasopressin (15-45 nM) inhibited the direct vasoconstrictor action of [Arg8]vasopressin but had no effect on the enhancement of the pressor response to methoxamine or electrical stimulation. The V1B receptor agonist [deamino-Cys1,beta-(3-pyridyl)-D Ala2,Arg8]vasopressin (100-1000 nM) and the V2 receptor agonist [deamino-Cys1,D Arg8]vasopressin (1-10 nM) were devoid of any pressor activity and did not potentiate methoxamine-evoked vasoconstriction. In contrast, [1 triglycyl,Lys8]vasopressin (100 - 1000 nM) potentiated the methoxamine responses without per se inducing vasoconstriction. In arteries precontracted with methoxamine (7.5 microM) pressor responses to [Arg8]vasopressin (3-10 nM) were not inhibited by a dose of SR 49,059 (3 nM) which abolished the peptide's vasoconstrictor effect under control conditions. These data show that the direct vasoconstrictor effect of [Arg8]vasopressin is mediated by V1A receptors while the enhancement of adrenoceptor-mediated pressor responses is insensitive to V1A, V1B, and oxytocin receptor antagonists and is not mimicked by selective agonists of V1B and V2 receptors. In conclusion, an unusual interaction of vasopressin with V1A receptors, or even the existence of a novel receptor subtype, has to be considered. PMID- 9863638 TI - Differential role of vasoactive prostanoids in porcine and human isolated pulmonary arteries in response to endothelium-dependent relaxants. AB - The pig is increasingly being used in medical research, both as a model of the human cardiovascular system, and as a possible source of organs for xenotransplantation. However, little is known about the comparative functions of the vascular endothelium between porcine and human arteries. We have therefore compared the effects of two endothelium-dependent vasorelaxants, acetylcholine (ACh) and the Ca2+-ATPase inhibitor, cyclopiazonic acid (CPA) on the porcine and human isolated pulmonary artery using isometric tension recording. ACh and CPA produced endothelium-dependent relaxations of both the human and porcine pulmonary arteries. In the porcine pulmonary artery, the cyclo-oxygenase inhibitor, flurbiprofen had no effect on relaxations to ACh (Emax: control 67.8+/ 8.8% versus 72.4+/-9.5% (n=11)) or CPA (Emax: control 79.6+/-5.0% versus 94.0+/ 10.6% (n=7)). The nitric oxide synthase inhibitor, L-NAME converted relaxations to both ACh and CPA into contractile responses (maximum response: ACh 30.0+/ 11.1% (n = 10); CPA 80.4+/-26.2% (n = 8) of U46619-induced tone). These contractile responses in the presence of L-NAME were abolished by flurbiprofen. In the human pulmonary artery, L-NAME and flurbiprofen partly attenuated relaxations to ACh (Emax: control: 45.1+/-12.1%; flurbiprofen: 33.4+/-13.5%; L NAME: 10.1+/-7.2%) and CPA (Emax: control: 78.1+/-5.5%; flurbiprofen: 69.6+/ 7.2%; L-NAME 37.9+/-10.7% of U46619-induced tone). These responses were abolished by the combination of both inhibitors. We have demonstrated that while the release of nitric oxide is important in responses to endothelium-dependent vasorelaxants in both human and porcine pulmonary arteries, in the human arteries, there is an important role for vasorelaxant prostanoids whilst in the porcine arteries, vasoconstrictor prostanoids are released. PMID- 9863639 TI - Inhibition of the rapid component of the delayed-rectifier K+ current by therapeutic concentrations of the antispasmodic agent terodiline. AB - Prolongation of the QT interval and malignant ventricular arrhythmia have been observed in patients administered terodiline for urinary incontinence. Since this adverse reaction might be caused by inhibition of delayed-rectifier K+ current (IK), we investigated whether clinically relevant (< or = 10 microM) concentrations of the drug modify IK in guinea-pig ventricular myocytes. Myocytes superfused with normal Tyrode's solution were pulsed from -40 mV to more positive test potentials (V) for 0.2 - 1 s to elicit tail IK on repolarization and measure tail IK-V relationships. IKr was distinguished from IKs by its sensitivity to the selective blocker E4031. Inhibition of IKr by 5 microM E4031 was completely occluded by pretreatment with 3 microM terodiline. In addition, action potential lengthening by E4031 in guinea-pig papillary muscles (29+/-3%) was abolished (3+/ 2%) (P<0.001) by terodiline pretreatment. Inhibition of IKr by terodiline appeared to be voltage-independent, and the parameters of the Hill equation describing the inhibition were IC50 = 0.7 microM and nH = 1.6. High concentrations of the drug also affect IKs; in experiments with K+-free Tyrode's, 10 microM terodiline inhibited tail IKs by 27+/-3% (n=5) (P< 0.001). These data suggest that QT lengthening at therapeutic concentrations of the drug (approximately equal to 1.5 microM) is primarily due to inhibition of IKr. Inhibition of other K+ currents such as IKs is likely to be important at higher concentrations. PMID- 9863640 TI - Impaired function of alpha-2 adrenoceptors in smooth muscle of mesenteric arteries from spontaneously hypertensive rats. AB - The alpha2-adrenoceptor function in mesenteric arteries of spontaneously hypertensive rats (SHR) was investigated by comparing membrane potential changes in response to adrenergic agonists in preparations from female SHR, Wistar-Kyoto (WKY) and normotensive Wistar rats (NWR). Resting membrane potential was found to be less negative in mesenteric arteries from SHR than in those from NWR and WKY. Apamin induced a decrease in the membrane potential of mesenteric artery rings without endothelium from NWR and WKY, but had no effects in those from SHR. Both UK 14,304 and adrenaline, in the presence of prazosin, induced a hyperpolarization that was significantly lower in de-endothelialized mesenteric rings from SHR than in those from NWR and WKY. In mesenteric rings with endothelium, however, similar hyperpolarization was observed in the three strains. In NWR mesenteric rings with endothelium the hyperpolarization induced by activation of alpha2-adrenoceptors was abolished by apamin, whereas in intact SHR mesenteric rings this hyperpolarization was slightly reduced by apamin and more efficiently reduced by Nomega-nitro-L-arginine. It is concluded that the activity of potassium channels coupled to alpha2-adrenoceptors is altered in the smooth muscle cells of SHR mesenteric arteries, contributing to their less negative membrane potential. On the other hand, the endothelial alpha2-receptors are functioning in mesenteric vessels from SHR and their stimulation induces a hyperpolarization mainly through the release of nitric oxide. PMID- 9863641 TI - The mechanism by which aminoglycoside antibiotics cause vasodilation of canine cerebral arteries. AB - The effects of aminoglycoside antibiotics were examined in canine cerebral arteries and in cultured cerebrovascular smooth muscle cells stimulated with oxyhemoglobin (OxyHb), a blood constituent which has been implicated in the pathogenesis of cerebrovascular spasm. In cerebral arterial rings precontracted with OxyHb (10 microM), the aminoglycosides caused a concentration-dependent decrease in isometric tension. The EC50s for the relaxation were 0.46+/-0.1 mM (n=6), 0.53+/-0.08 mM (n=12), 1.6+/-0.3 mM (n=7) and 3.9+/-0.5 mM (n=5) for neomycin, gentamicin, streptomycin and kanamycin, respectively. This order of potency corresponds approximately to the number of positive charges in the molecules. The aminoglycosides also inhibited the contractions to prostaglandin F2alpha (1 microM) and depolarizing concentrations of potassium chloride (60 mM). The order of potency was neomycin > gentamicin > streptomycin > kanamycin. The relaxation was maintained in vascular preparations denuded of endothelium. Neomycin (5 mM) abolished the Ca2+-independent contraction to PGF2alpha. In Fura 2-loaded cerebrovascular smooth muscle cells, OxyHb (1 microM) significantly enhanced the concentration of intracellular calcium ([Ca2+]i) by 330%. The administration of neomycin, gentamicin, kanamycin and streptomycin in concentrations corresponding to the EC50 from contractility studies, reduced the effects of OxyHb on [Ca2+]i by about 50% to 221+/-35 nM (n=7), 270+/-31 nM (n=7), 229+/-33 nM (n = 6) and 240+/-6 nM (n = 5), respectively. These results suggests that the effects of the aminoglycosides on the OxyHb-induced contraction and the long-term increase in [Ca2+]i, may arise from several effects, including inhibition of PLC, protection of calcium extrusion mechanisms, and interference with the process of [Ca2+]i accumulation. PMID- 9863642 TI - Comparison of two soluble guanylyl cyclase inhibitors, methylene blue and ODQ, on sodium nitroprusside-induced relaxation in guinea-pig trachea. AB - To clarify further the role of cyclic GMP in mediating the relaxant response in guinea-pig trachea induced by sodium nitroprusside (SNP), the effects of soluble guanylyl cyclase inhibitors, methylene blue and 1H-[1,2,4]oxadiazolo[4,3, a]quinoxalin-1-one (ODQ) on SNP-induced muscle relaxation and cyclic GMP accumulation were determined. SNP (0.3-100 microM) evoked a concentration dependent relaxation of guinea-pig isolated tracheas precontracted with 0.3 microM carbachol. Preincubation of the preparations with methylene blue (10, 30 and 100 microM) resulted in a slight but concentration-dependent prevention of the relaxant response to SNP. In contrast, the relaxation to SNP was extensively prevented by 3 microM ODQ and almost abolished by 10 microM ODQ. SNP (30 microM) induced a significant elevation of cyclic GMP accumulation (from 1.34+/-0.14 to 5.39+/-0.28 pmol mg(-1) protein, n= 5; P<0.001), which was partially attenuated by 100 microM methylene blue (3.11+/-0.51 pmol mg(-1) protein, n=5; P<0.05), whereas completely abolished by 10 microM ODQ (1.31+/-0.28 pmol mg(-1) protein, n = 5; P<0.001). Methylene blue, but not ODQ and Nomega-nitro-L-arginine methyl ester (L-NAME), caused a concentration-dependent contraction in the tracheal preparation. The tension produced by 100 microM methylene blue was 41.8+/-4.3% (0.3 microM carbachol as 100%; n = 12). Moreover, the non-selective muscarinic receptor antagonist atropine and the M3-selective antagonist 4-diphenylacetoxy-N methylpiperidine methiodine greatly inhibited the contractile effect evoked by methylene blue (100 microM). In conclusion, this study provides substantial evidence that SNP-induced muscle relaxation in guinea-pig trachea is completely via a cyclic GMP-dependent mechanism. Furthermore, ODQ, but not methylene blue, will likely become an important tool in differentiating between cyclic GMP dependent and -independent effects of nitric oxide. PMID- 9863643 TI - Characterization of the relaxant action of urocortin, a new peptide related to corticotropin-releasing factor in the rat isolated basilar artery. AB - In addition to its well established neuroendocrine and neurotransmitter effects, corticotropin releasing factor (CRF) exerts a potent vasorelaxant action. Recently, a CRF-related peptide, urocortin, has been identified in the mammalian brain. In the present study, the cerebral vasomotor action of this peptide and the mechanism underlying its relaxant effect are characterized. Ring segments obtained from the rat basilar artery were used for measurement of isometric force. The relaxant action of urocortin, CRF and sauvagine was studied in segments with a functionally intact endothelium. In segments precontracted with prostaglandin F2alpha, urocortin, CRF and sauvagine induced concentration-related relaxation. The order of potency was as follows (pD2+/-s.e.m. given in brackets): urocortin (9.32+/-0.07) > sauvagine (9.08+/-0.08) > CRF (7.50+/-0.07). Complete relaxation was achieved with each agonist. Relaxation was not affected by removal of the endothelium but was markedly attenuated in segments precontracted with 50 mM K+ Krebs solution. The relaxant effect of urocortin was inhibited by astressin in an apparently competitive manner. A pA2 value of 7.52 was estimated for astressin. Inhibition of urocortin-induced relaxation was also observed in the presence of the adenylate cyclase inhibitor SQ22536 (pD2 in the presence of 300 microM SQ22536, 9.36+/-0.05) and the K+ channel blockers tetraethylammonium (10 mM; pD2, 8.65+/-0.07), iberiotoxin (100 nM; pD2, 8.88+/-0.08) and apamin (10 nM; pD2, 8.94+/-0.07). However, the inhibitory actions of SQ22536 and apamin or iberiotoxin were not additive. The results suggest that urocortin induces relaxation of cerebral arteries by activating CRF-R2 receptors present in the vascular wall. Relaxation appears to be mediated by adenylate cyclase stimulation and activation of Ca2+-dependent K+ channels. PMID- 9863644 TI - The central action of the 5-HT2 receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2 aminopropane (DOI) on cardiac inotropy and vascular resistance in the anaesthetized cat. AB - Experiments were carried out to determine the effects of the application of the selective 5-HT2 receptor agonist DOI intravenously (in the presence of the peripherally acting 5-HT2 receptor antagonist, BW501C67, 1 mg kg(-1), i.v.) or to the 'glycine sensitive area' of the ventral surface (30 microg each side) on the left ventricular inotropic (left ventricular dP/dt max) and vascularly isolated hindlimb responses in anaesthetized cats. For the ventral surface experiments, NMDA (10 microg each side) was applied to act as a positive control. In all experiments heart rate and mean arterial blood pressure were held constant to exclude any secondary effects caused by changes in these variables. DOI (n=6) i.v or on the ventral surface had no effect on left ventricular dP/dt max but caused a significant increase in hindlimb perfusion pressure of 40+/-9 and 50+/-14 mmHg, respectively. Respiration was unaffected. NMDA (n=6), applied to the ventral surface, caused significant increases in both left ventricular dP/dt max and hindlimb perfusion pressure of 1,950+/-349 mmHg s(-1) and 69+/-17 mmHg respectively, with no associated change in left ventricular end-diastolic pressure. The amplitude of respiratory movements increased. It is concluded that activation of 5-HT2 receptors at the level of the rostral ventrolateral medulla (RVLM) excites sympathetic premotor neurons and/or their antecedents controlling hindlimb vascular resistance but not those controlling the inotropic effects on the left ventricle. PMID- 9863645 TI - The activation of nitric oxide synthase by copper ion is mediated by intracellular Ca2+ mobilization in human pulmonary arterial endothelial cells. AB - The aim of the study was to elucidate the vasodilatory mechanism due to Cu2+ by assessing nitric oxide (NO) production as determined by NOx (NO, NO2-, and NO3-) that is released from human pulmonary arterial endothelial cell (HPAEC) monolayers using a NO chemiluminescence analyzer, and also to assess Ca2+ movement using 45Ca and fura 2 in HPAEC. Cu2+ (10(-6)-10(-4) M) significantly increased NO production in a dose-dependent manner when extracellular Ca2+ was present. 45Ca influx into the adherent cells was dose-dependently enhanced by Cu(2+) (10(-6)-10(-4) M), but not by Mn(2+), Zn(2+) or Fe(2+). [Ca2+]i, measured by monitoring the fluorescence changes of fura 2, was significantly elevated in the presence of Cu2+. The increase in [Ca2+]i induced by Cu2+ was inhibited by either diethyldithiocarbamate (DDC) or the depletion of extracellular Ca2+. The dihydropyridine receptor agonist, BayK8644, significantly attenuated the Cu2+ induced increase in [Ca2+]i in a dose dependent manner and nitrendipine or nifedipine, the dihydropyridine receptor antagonists, dose-dependently inhibited a Cu2+-induced increase in [Ca2+]i. These results suggest that Cu2+ activates eNOS through the mechanism of [Ca2+]i elevation due to Ca2+ influx into HPAEC and that the Cu2+-induced [Ca2+]i elevation in HPAEC is likely due to activation of the dihydropyridine-like receptors. PMID- 9863646 TI - Functional cross-talk between endothelial muscarinic and alpha2-adrenergic receptors in rabbit cerebral arteries. AB - Interactions between two classes of receptors have been observed in several cell lines and preparations. The aim of this work was to assess the impact of simultaneous stimulation of endothelial muscarinic and alpha2-adrenergic receptors (alpha2-AR) on vascular reactivity. Rabbit middle cerebral arteries were isolated and changes in isometric tension were recorded in the presence of indomethacin. Inhibition of nitric oxide (NO) synthase with Nomega-nitro-L arginine (L-NOARG, 100 micromol l(-1)) revealed alpha-AR-dependent contractions. Pre-addition of acetylcholine (ACH, 1 micromol l(-1)) augmented oxymetazoline (OXY, 10 micromol l(-1), alpha2-AR agonist)-, but decreased phenylephrine (PE, 10 micromol(-1), alpha1-AR agonist)-induced contraction (P<0.05). The effects of ACH were endothelium-dependent. Vessels were precontracted with 40 mmol l(-1) KCl physiological salt solution (PSS) in the absence of L-NOARG, or PE or OXY in the presence of L-NOARG. In the presence of high external K+ or PE, ACH induced a potent relaxation (P<0.05). In the presence of OXY, however, ACH mediated contraction (P<0.05). After pertussis toxin (PTX, inactivator of Galpha(i/o) proteins) pre-treatment, alpha2-AR-dependent contractions were abolished. Forty mmol l(-1) KCl-PSS induced contraction was not altered by PTX whereas ACH-induced relaxation was augmented (P<0.05). To investigate if endothelin-1 (ET-1) intervened in the endothelium-dependent contractile response to ACH in the presence of OXY-dependent tone, vessels were incubated in the presence of BQ123 (1 micromol l(-1)), an ETA receptor antagonist. OXY-mediated tone was not affected by BQ123; however, ACH-induced contraction was reversed to a relaxation (P<0.05). These data indicate that activation of endothelial alpha2-AR triggers an endothelium-dependent, ET-1 mediated, contraction to ACH. This suggests that activation of alpha2-AR affects muscarinic receptor/G protein coupling leading to an opposite biological effect. PMID- 9863647 TI - Identification and characterization of an endogenous P2X7 (P2Z) receptor in CHO K1 cells. AB - CHO-K1 cells were examined for their cellular responses to the P2 receptor agonist, 2'- and 3'-O-(4-benzoylbenzoyl)-ATP (DbATP), and for the presence of mRNA for P2X receptors. Reverse transcriptase-polymerase chain reactions, using primers directed against the rat P2X subunits, detected the presence of P2X7 but not P2X1-P2X6 subunits. DbATP (EC50 approximately equal to 100 microM) evoked non desensitizing inward currents which reversed at approximately equal to 0 mV, suggesting activation of a non-selective cation channel. ATP also evoked inward currents but was less potent than DbATP. DbATP also stimulated the accumulation of 45calcium (45Ca2+) and the DNA binding dye, YO-PRO-1, in CHO-KI cells. Both responses were inhibited by NaCl and MgCl2. In 280 mM sucrose buffer, 45Ca2+ accumulation was measurable within 10-20 s of agonist addition, whereas YO-PRO-1 accumulation was only detectable after 8 min. ATP and ATPgammaS were also agonists but were less potent than DbATP, while UTP, 2-methylthio ATP, ADP and (alphabeta)methylene ATP were inactive at concentrations up to 100 microM. DbATP increased lactate dehydrogenase release from CHO-K1 cells, suggesting cell lysis, although this effect was only pronounced after 60-90 min. These data suggest that CHO-K1 cells express an endogenous P2X7 receptor which can be activated by DbATP to cause a rapid inward current and accumulation of 45Ca2+. Prolonged receptor activation results in a delayed, increased permeability to larger molecules such as YO-PRO-1 and ultimately leads to cell lysis. Importantly, the presence of an endogenous P2X7 receptor should be considered when these cells are used to study recombinant P2X receptors. PMID- 9863648 TI - Mechanism of ET(A)-receptor stimulation-induced increases in intracellular Ca2+ in SK-N-MC cells. AB - The mechanism underlying endothelin-1 (ET-1)-induced increases in intracellular Ca2+ concentrations in the human neuroblastoma cell-line SK-N-MC was investigated. ET-receptor agonists increased inositol phosphate (IP)-formation (assessed as accumulation of total [3H]-IPs in [3H]-myo-inositol prelabelled cells) and intracellular Ca2+ (assessed by the FURA-2 method) with an order of potency: ET-1 > sarafotoxin 6b (S6b)> ET-3 = S6c; the ETA-receptor antagonist BQ 123 inhibited both responses with apparent pKi-values of 8.3 and 8.6, respectively, while the ETB-receptor antagonist BQ-788 did not. Pretreatment of the cells with pertussis toxin (PTX, 500 ng ml(-1) overnight) reduced ET-1 induced Ca2+ increases by 46+/-5%, but rather enhanced ET-1-induced IP-formation. Chelation of extracellular Ca2+ by 5 mM EGTA did not affect ET-1-induced IP formation. However, in the presence of 5 mM EGTA or SKF 96365, an inhibitor of receptor mediated Ca2+ influx (1.0-3.0 x 10(-5) M) ET-1-induced Ca2+ increases were inhibited in normal, but not in PTX-treated cells. [125I]-ET-1 binding studies as well as mRNA expression studies (by RT-PCR) detected only ETA receptors whereas expression of ETB-receptor mRNA was marginal. ET-1 (10(-8) M) inhibited isoprenaline-evoked cyclic AMP increases; this was antagonized by BQ 123, not affected by BQ-788 and abolished by PTX-treatment. We conclude that SK-N MC cells contain a homogeneous population of ETA-receptors that couple to IP formation and inhibition of cyclic AMP formation. Stimulation of these ETA receptors increases intracellular Ca2+ by at least two mechanisms: a PTX insensitive IP-mediated Ca2+ mobilization from intracellular stores and a PTX sensitive influx of extracellular Ca2+. PMID- 9863649 TI - Effect of nimesulide and indomethacin on contractility and the Ca2+ channel current in myometrial smooth muscle from pregnant women. AB - The non-steroidal anti-inflammatory drug (NSAID) indomethacin inhibits both constitutive and inducible forms of cyclo-oxygenase (COX-1 and COX-2, respectively), while nimesulide is a selective COX-2 inhibitor. Uterine COX-2 is upregulated before and during term and pre-term labour, and prostaglandins play a crucial role in parturition. We therefore evaluated the effects of these drugs on myometrial contractility and the voltage-gated Ca2+ channel current in tissue strips and isolated human myometrial smooth muscle cells (HMSMC) from myometrial biopsies taken with informed consent from women undergoing caesarean section at term (not in labour). Nimesulide and indomethacin caused almost complete inhibition of spontaneous myometrial contractions at concentrations of 100 and 300 microM, respectively. The Ca2+ channel current was inhibited in a concentration-dependent manner by both drugs, with a 40% reduction of the current at 100 microM nimesulide and 300 microM indomethacin. Nimesulide also accelerated the decay of the Ca2+ channel current. The inhibition of the Ca2+ channel current by 100 microM nimesulide and 300 microM indomethacin was unaffected by the presence of either PGF2alpha or PGE2 (30 microM), and was of similar magnitude whether 10 mM Ba2+ or 1.5 mM Ca2+ was used as the charge carrier. The concentrations of indomethacin and nimesulide required to suppress spontaneous contractility in human pregnant myometrium were much higher than those necessary to inhibit prostaglandin production. The results suggest that both nimesulide and indomethacin inhibit myometrial contractility via mechanisms independent of cyclo oxygenase inhibition. Blockade of the Ca2+ current may contribute to this effect. PMID- 9863651 TI - Priming and induction of eosinophil trafficking in guinea-pig cutaneous inflammation by tumour necrosis factor alpha. AB - Tissue eosinophilia is a hallmark of allergic and parasitic diseases. Priming mechanisms may play an important role in mediating the process of eosinophil accumulation in these conditions. We have previously shown that blockade of tumour necrosis factor alpha (TNFalpha) inhibited the capacity of lipopolysaccharide to prime skin sites for chemoattractant-induced eosinophil recruitment. The present study was carried out to investigate the capacity of TNFalpha to prime an inflammatory site for enhanced eosinophil accumulation. Initial experiments investigated the capacity of TNFalpha itself to induce eosinophil accumulation. Intradermal injection of murine TNFalpha (10-300 ng per site) in the guinea-pig induced significant accumulation of 111In-eosinophils. Kinetic studies showed the response to be delayed in onset and inhibited by cycloheximide, consistent with a dependency on protein synthesis. Trafficking of 111In-eosinophils to sites treated for 2 h with TNFalpha (10-100 ng per site) was inhibited by monoclonal antibodies (mAbs) against beta2 or alpha4 integrins. Intradermal injection of a low dose (3 ng) of TNFalpha (which by itself had no significant effect on eosinophil trafficking) prior to chemoattractants or antigen in sensitized skin sites, induced significant priming of eosinophil accumulation. Recruitment of both 111In-eosinophils and endogenous eosinophils was enhanced. Trafficking to TNFalpha-primed responses was dependent on protein synthesis and beta2 integrins. In contrast, the alpha4 integrin mAb failed to inhibit the TNFalpha primed response. Thus, TNFalpha can induce and also prime eosinophil recruitment in guinea-pig skin. Our results provide further evidence that this cytokine may be an important mediator of allergic- or parasite-induced eosinophilic inflammation. PMID- 9863650 TI - Persistent nicotinic blockade by chlorisondamine of noradrenergic neurons in rat brain and cultured PC12 cells. AB - Chlorisondamine (CHL) blocks behavioural responses to nicotine for several weeks or months in rats. Persistent blockade has also been demonstrated ex vivo, in assays of nicotine-evoked striatal dopamine release. Central administration of [3H]-CHL leads to long-term retention of radiolabel in nigrostriatal dopaminergic neurons and in few other cell groups. We investigated whether an analogous blockade also occurs in noradrenergic neurons in the brain and in cultured pheochromocytoma (PC12) cells, which have a similar noradrenergic phenotype. Administration of CHL (10 mg kg(-1) s.c. or 10 microg i.c.v.), 21 days prior, resulted in a near-total block of nicotine-evoked release of hippocampal [3H] noradrenaline ([3H]-NA) from superfused rat synaptosomes; NMDA-evoked [3H]-NA release was unaffected. Three weeks after administration of [3H]-CHL (10 microg i.c.v.), preferential accumulation of radiolabel was observed in the locus coeruleus, which provides the entire noradrenergic innervation to hippocampus, as well as in previously noted structures. In rat pheochromocytoma (PC12) cells, nicotine evoked [3H]-NA release (EC50 approximately 30 microM). This effect was blocked by co-incubation with mecamylamine (10 microM) or CHL (1 microM) but was not affected by alpha-bungarotoxin. As in the hippocampus, the nicotinic agonist cytisine was at least as efficacious as nicotine. Acute exposure of PC12 cells to CHL 10 or 100 microM (but not 1 microM), followed by 90 min wash-out, almost completely blocked release evoked by 30 microM nicotine. More prolonged (24 h) exposure to CHL 100 microM (but not 1 or 10 microM), followed by 3 days of wash out, partially inhibited release evoked by nicotine, leaving responses to high K+ unchanged. A significant (30%) reduction was also seen 5 days after exposure. We conclude that persistent nicotinic blockade by CHL is neither restricted to mesostriatal dopamine neurons, nor to the CNS, nor to neurons possessing the same nicotinic receptor pharmacology. In addition, the persistent blockade does not appear to result from an acute blocking action, but may be dependent upon intracellular accumulation of the antagonist. PMID- 9863652 TI - The antihypertensive profile of the angiotensin AT1 receptor antagonist, GR138950, and the influence of potential homeostatic compensatory mechanisms in renal hypertensive rats. AB - The cardiovascular profile of the angiotensin AT1 receptor antagonist, GR138950, and the influence of potential compensatory homeostatic mechanisms on this profile, were investigated in renal artery ligated hypertensive (RALH) rats. GR138950 caused a marked reduction in blood pressure associated with immediate tachycardia in conscious RALH rats. The antihypertensive action of GR138950 appeared biphasic; an immediate fall in blood pressure, which plateaued within 1 h, and which was followed by a further slow decline that reached maximum between 5-7 h after administration. The tachycardia caused by GR138950 was attenuated by atenolol and was abolished by combined pretreatment with atenolol and atropine methyl nitrate. However, the antihypertensive profile of GR138950 was unchanged by these pretreatments. The resting blood pressure and the antihypertensive effect of GR138950, in RALH rats, were unaffected by the vasopressin V1 receptor antagonist, [beta-mercapto-beta,beta-cyclopentamethylene propionyl(1)-O-Me Tyr2,Arg8]-vasopressin. Thus, vasopressinergic mechanisms are not involved in either maintaining blood pressure in RALH rats, or in compensating for the fall in blood pressure caused by GR138950. In anaesthetized RALH rats, GR138950 caused a marked fall in blood pressure that was accompanied by an increase in heart rate along with sustained increases in renal and splanchnic sympathetic nerve activity. In summary, the biphasic fall in blood pressure evoked by GR138950 in RALH rats can not be explained on the basis of changes in autonomic control of the heart, alteration of vasopressin-mediated vasoconstrictor mechanisms or overall suppression of central sympathetic outflow. Rather, increased vasoconstrictor tone might serve to oppose the initial fall in blood pressure. PMID- 9863653 TI - Venous versus arterial actions of diethylamine/nitric oxide (DEA/NO) complex and S-nitroso-N-acetylpenicillamine (SNAP) in vivo. AB - We studied the effects of diethylamine/NO complex (DEA/NO) and S-nitroso-N acetylpenicillamine (SNAP), relative to those of sodium nitroprusside (SNP) and nitroglycerin (NTG), on mean arterial pressure (MAP), mean circulatory filling pressure (MCFP), arterial resistance (Ra), venous resistance (Rv), heart rate (HR), cardiac output (CO) and stroke volume (SV) in groups of Inactin anaesthetized rats pre-treated with i.v. mecamylamine (3.7 micromol kg(-1)) and noradrenaline (6.8 nmol kg(-1) min(-1)). Doses of each that reduced MAP by 30%, 80% and the lowest dose that maximally reduced MAP were examined to allow a comparison of the compounds' dilator actions at equivalent effective depressor doses. DEA/NO (4, 32 and 256 microg kg(-1) min(-1)), SNAP (4, 32 and 256 microg kg(-1) min(-1)) and SNP (8, 32 and 128 microg kg(-1) min(-1)) caused similar dose dependent reductions in MAP and Ra, and increases in CO and SV. NTG (0.2, 0.8 and 6.4 microg kg(-1) min(-1)) dose-dependently reduced Ra, and increased CO and SV, but lowered MAP only at the highest dose. DEA/NO, SNAP and SNP but not NTG lowered MCFP with efficacy: DEA/NO > SNAP > SNP. All four drugs reduced Rv with efficacy: DEA/NO approximately equal to SNAP > SNP approximately equal to NTG. Therefore, all compounds lowered Ra and Rv. DEA/NO, SNAP and SNP but not NTG reduced MCFP. The pharmacological profiles of DEA/NO and SNAP resemble SNP more than NTG. PMID- 9863654 TI - Evidence that platelets promote tube formation by endothelial cells on matrigel. AB - The involvement of platelets in neovascularization was investigated in the matrigel tube formation assay, an in vitro model of angiogenesis. Platelets promoted the formation of capillary-like structures (expressed as relative tube area) number- and time-dependently. Relative tube area increased from 0.98+/-0.02 (n = 8) in the presence of 6.25 x 10(4), to 3.21+/-0.12 (n=8) in the presence of 10(6) platelets/well compared to 0.54+/-0.04 (n=8) in their absence. This increase was unaffected by acetyl salicylic acid (ASA), apyrase, and hirudin. Photographs from representative experiments, showed that platelets adhered along the differentiating endothelium. Addition of alpha-thrombin (0.1-1 i.u. ml(-1)), the nitric oxide (NO) donor sodium nitroprusside (SNP; 1-100 microM) or the NO synthase inhibitor, L-NG-arginine-methylester (L-NAME, 30-300 microM) to the assay, had no effect on tube formation compared to that seen with platelets alone. Neuraminidase (0.01 i.u./10(7) platelets), which strips sialic acid residues from membrane glycoproteins, abolished the promoting effect of platelets on tube formation. The relative tube area in the presence of neuraminidase treated platelets was 0.81+/-0.03 (n = 8), in the presence of untreated platelets 1.69+/-0.09, P<0.001 (n=8) and in the absence of platelets, 0.80+/-0.04 (n=8). The tetrapeptide Arg-Gly-Asp-Ser (RGDS; 20-200 microM) which inhibits von Willebrand factor, fibrinogen and fibronectin-mediated adhesion, had no effect on the promoting effect of platelets on tube formation. These results indicate that platelets promote angiogenesis in vitro. This effect is largely independent from activation by alpha-thrombin, is not modified by manipulating NO and prostaglandin metabolism and proceeds possibly through adhesion of the platelets to the differentiating endothelium. PMID- 9863655 TI - LU 73068, a new non-NMDA and glycine/NMDA receptor antagonist: pharmacological characterization and comparison with NBQX and L-701,324 in the kindling model of epilepsy. AB - The aim of this study was to assess whether a drug which combines an antagonistic action at both NMDA and non-NMDA receptors offers advantages for treatment of epileptic seizures compared to drugs which antagonize only one of these ionotropic glutamate receptors. The novel glutamate receptor antagonist LU 73068 (4,5-dihydro-1-methyl-4-oxo-7-trifluoromethylimidazo[1,2a]quinoxal ine-2-carbonic acid) binds with high affinity to both the glycine site of the NMDA receptor (Ki 185 nM) and to the AMPA receptor (Ki 158 nM). Furthermore, binding experiments with recombinant kainate receptor subunits showed that LU 73068 binds to several of these subunits, particularly to rGluR7 (Ki 104 nM) and rGluR5 (Ki 271 nM). In comparison, the prototype non-NMDA receptor antagonist NBQX (2,3-dihydroxy-6 nitro-7-sulphamoyl-benzo[f]quinoxaline) binds with high affinity to AMPA receptors only. Both NBQX and LU 73068 were about equieffective after i.p. injection in mice to block lethal convulsions induced by AMPA or NMDA. In the rat amygdala kindling model of temporal lobe epilepsy, LU 73068 dose-dependently increased the focal seizure threshold (afterdischarge threshold, ADT). When rats were stimulated with a current 20% above the individual control ADT, LU 73068 completely blocked seizures with an ED50 of 4.9 mg kg(-1). Up to 20 mg kg(-1), only moderate adverse effects, e.g. slight ataxia, were observed. NBQX, 10 mg kg( 1), and the glycine/NMDA site antagonist L-701,324 (7-chloro-4-hydroxy-3-(3 phenoxy)phenyl-quinoline-2(1H)one), 2.5 or 5 mg kg(-1), exerted no anticonvulsant effects in kindled rats when administered alone, but combined treatment with both drugs resulted in a significant ADT increase. The data indicate that combination of glycine/NMDA and non-NMDA receptor antagonism in a single drug is an effective means of developing a potent and effective anticonvulsant agent. PMID- 9863656 TI - Adenosine A1 receptor-mediated excitation of nociceptive afferents innervating the normal and arthritic rat knee joint. AB - We tested the hypothesis that adenosine excites nociceptive primary afferents innervating the knee joint. Neuronal recordings were made from fine nerve filaments innervating the knee joint in rats anaesthetized with pentobarbitone. Drugs were injected close-arterially (i.a.) or into the articular space (i.art.). We studied normal and chronically inflamed arthritic joints, the latter 14-21 days after a single intra-articular injection of Freund's Complete Adjuvant, performed under halothane anaesthesia. Adenosine injected i.a. caused delayed (approximately 10 s) excitation of the majority of polymodal C-fibre afferents, and had similar effects when injected directly into the joint. Adenosine triphosphate (ATP) had biphasic effects on discharge, a fast (<1 s) excitation was followed by a delayed increase similar to that seen with adenosine. The adenosine A1 receptor agonists N6-cyclopentyladenosine (CPA) and N-[(1S,trans)-2 hydroxypentyl] adenosine (GR79236) also excited the C-fibre afferents. The A1 antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) antagonized the responses evoked by adenosine, CPA, and the delayed increase seen after ATP, indicating that excitation of the nociceptive afferents was mediated via adenosine A1 receptors. Adenosine and ATP evoked delayed excitatory effects of similar magnitude, regardless of whether or not the knee joint was chronically inflamed. The increased basal discharge observed in arthritic joints was unaffected by DPCPX, which implies that the increase in spontaneous activity associated with arthritis is unlikely to involve tonically released adenosine. The results support the hypothesis that adenosine excites primary afferent nociceptive nerve terminals in the rat knee joint, an effect mediated by adenosine A1 receptors. ATP, adenosine, and A1 receptors may play a role in generating the peripheral nociceptive (pain) signal. PMID- 9863657 TI - Analysis of an H1 receptor-mediated, zinc-potentiated vasoconstrictor action of the histidyl dipeptide carnosine in rabbit saphenous vein. AB - The contractile action of the dipeptide carnosine (beta-alanyl-L-histidine), active as a Zn.carnosine complex (Zn. Carn), was investigated in isolated rings of rabbit saphenous vein (RSV) and was found to be antagonized by the H1 antagonist mepyramine. Mepyramine-sensitive, histamine-induced contractures in RSV, were smaller (73+/-0.1%) and less well sustained than carnosine-induced contractures. Schild plot values for mepyramine antagonism were, for carnosine induced contractures; pA2 = 7.97+/-0.12, slope= 1.33+/-0.06 (r = 0.793) and for histamine-induced contractures; pA2 = 8.48+/-0.07, slope = 0.63+/-0.05, r = 0.957). Serotonergic antagonists methiothepin and ketanserin, antagonize both carnosine- and histamine-induced contractures in RSV, probably reflecting coincidental inhibition at the H1-receptor. Carnosine, with Zn present, can inhibit the H1-specific binding of [3H]-mepyramine to isolated guinea-pig cerebellar membranes (log IC50s - 2.78+/-0.02, -3.93+/-0.03 and -4.64+/-0.03 at 10, 30 and 80 microM Zn respectively; values corrected for the Zn-specific inhibition which has a logIC50 of -4.20). In the radioligand binding assay, the effect of carnosine can be described as a function of Zn. Carn concentration with an apparent logIC50 of -5.61. This value is consistent with that obtained from the functional studies on RSV. Histamine-induced contractures have an indomethacine-sensitive component (27.2+/-8.3% of control response), not apparent with carnosine-induced contractures. Like histamine, carnosine evoked an H2 mediated (cimetidine-sensitive) relaxation in the presence of mepyramine, but was less potent (10.8+/-3.1% residual tension at 10 mM carnosine compared with 13.4+7.5% at 0.1 mM histamine). Carnosine, like mepyramine, can 'reveal' the H2 mediated relaxation of histamine providing further evidence that carnosine binds at the H1 receptor. We conclude that carnosine can act at the smooth muscle H1 receptor to provoke vasoconstriction and that it also has the potential to act at H1-receptors in CNS. PMID- 9863658 TI - Potentiating action of MKC-242, a selective 5-HT1A receptor agonist, on the photic entrainment of the circadian activity rhythm in hamsters. AB - Serotonergic projections from the midbrain raphe nuclei to the suprachiasmatic nuclei (SCN) are known to regulate the photic entrainment of circadian clocks. However, it is not known which 5-hydroxytryptamine (5-HT) receptor subtypes are involved in the circadian regulation. In order to verify the role of 5-HT1A receptors, we examined the effects of 5-?3-[((2S)-1,4-benzodioxan-2 ylmethyl)amino]-propoxy?-1,3-b enzodioxole HCl (MKC-242), a selective 5-HT1A receptor agonist, on photic entrainment of wheel-running circadian rhythms of hamsters. MKC-242 (3 mg kg(-1), i.p.) significantly accelerated the re entrainment of wheel-running rhythms to a new 8 h delayed or advanced light-dark cycle. MKC-242 (3 mg kg(-1), i.p.) also potentiated the phase advance of the wheel-running rhythm produced by low (5 lux) or high (60 lux) intensity light pulses. In contrast, 8-hydroxydipropylaminotetralin (8-OH-DPAT)(5 mg kg(-1), i.p.), a well known 5-HT1A/5-HT7 receptor agonist, only suppressed low intensity (5 lux) light-induced phase advances. The potentiating actions of MKC-242 on light pulse-induced phase advances were observed even when injected 20 or 60 min after the light exposure. The potentiating action of MKC-242 was antagonized by WAY100635, a selective 5-HT1A receptor blocker, but not by ritanserin, a 5-HT2/5 HT7 receptor blocker, indicating that MKC-242 is activating 5-HT1A receptors. Light pulse-induced c-fos expression in the SCN and the intergeniculate leaflet (IGL) were unaffected by MKC-242 (3 mg kg(-1), i.p.). HPLC analysis demonstrated that MKC-242 (3 mg kg(-1), i.p.) decreased the 5-HIAA content in the SCN. The present results suggest that presynaptic 5-HT1A receptor activation may be involved in the potentiation of photic entrainment by MKC-242 in hamsters. PMID- 9863659 TI - Characterization of a prostanoid EP3-receptor in guinea-pig aorta: partial agonist action of the non-prostanoid ONO-AP-324. AB - Contraction of guinea-pig isolated aorta induced by the prostaglandin E analogue sulprostone (1-400 nM) has a lower maximum response (40%) than that of phenylephrine or U-46619 (TP-receptor agonist). A prostanoid EP3-receptor subtype is involved based on agonist potency ranking: equi-effective molar ratios (EMR) are sulprostone (EC50 approximately equal to 23 nM) 1.0, SC-46275 0.11, misoprostol 2.2, gemeprost 3.3, PGE2 5.4, 17-phenyl PGE2 6.0, GR-63799 8.9. GR 63799, which contains a bulky ester group, is relatively more potent on neuronal EP3 preparations than on the aorta. ONO-AP-324, a relative of the non-prostanoid prostacyclin mimetic series, behaves as an EP3 partial agonist on the aorta, inhibiting sulprostone responses but acting synergistically (in a similar manner to sulprostone) with phenylephrine; it may be a useful pharmacological tool for studying EP3-receptors. Sulprostone contractions are markedly suppressed in zero Ca2+ bathing fluid containing either 2 mM EDTA or 50 microM EGTA, and by Cd2+ (500 microM), but are usually unaffected by nifedipine (0.3 microM) and verapamil (4.44 microM). Influx of Ca2+, but not through L-type Ca2+-channels, appears to be the major contractile mechanism. The guinea-pig aorta is a valuable addition to the vascular EP3 preparations available and may increase our knowledge of the mechanisms whereby Gi-coupled receptors mediate vasoconstriction (c.f. 5-HT1B/D- and alpha2-receptors). The possibility of certain EP3 agonists distinguishing EP3 receptor isoforms is discussed. PMID- 9863660 TI - Exogenous and endogenous catecholamines inhibit the production of macrophage inflammatory protein (MIP) 1 alpha via a beta adrenoceptor mediated mechanism. AB - Noradrenaline (NA) and adrenaline (Ad) are modulators of cytokine production. Here we investigated the role of these neurotransmitters in the regulation of macrophage inflammatory protein (MIP)-1alpha expression. Pretreatment of RAW 264.7 macrophages with NA or Ad decreased, in a concentration-dependent manner (1 nM-100 microM), MIP-1alpha release induced by bacterial lipopolysaccharide (LPS 10 ng ml(-1) LPS). The effect of NA was reversed by the selective beta adrenoceptor antagonist propranolol (10 microM), but not by the alpha adrenoceptor antagonist phentolamine (10 microM). In the concentration range of 10 nM-10 microM, isoproterenol, a beta-adrenoceptor agonist, but not phenylephrine (a selective alpha1-adrenoceptor agonist) or UK-14304 (a selective alpha2-adrenoceptor agonist) mimicked the inhibitory effects of catecholamines on MIP-1alpha production. Increases in intracellular cyclic adenosine monophosphate, elicited either by the selective type IV phosphodiesterase inhibitor rolipram (0.1 - 10 microM), or by prostaglandin E2, (10 nM-10 microM) decreased MIP-1alpha release, suggesting that increased cyclic AMP may contribute to the suppression of MIP-1alpha release by beta-adrenoceptor stimulation. Northern blot analysis demonstrated that NA (100 nM-10 microM), Ad, isoproterenol, as well as rolipram (100 nM-10 microM) decreased LPS-induced MIP-1alpha mRNA accumulation. NA and Ad (1-100 microM) also decreased MIP-1alpha production in thioglycollate-elicited murine peritoneal macrophages. Pretreatment of mice with either isoproterenol (10 mg kg(-1), i.p.) or rolipram (25 mg kg(-1), i.p.) decreased LPS-induced plasma levels of MIP-1alpha, while propranolol (10 mg kg(-1), i.p.) augmented the production of this chemokine, confirming the role of a beta-adrenoceptor mediated endogenous catecholamine action in the regulation of MIP-1alpha production in vivo. Thus, based on our data we conclude that catecholamines are important endogenous regulators of MIP-1alpha expression in inflammation. PMID- 9863661 TI - Unilateral injection of calcitonin gene-related peptide (CGRP) induces bilateral oedema formation and release of CGRP-like immunoreactivity in the rat hindpaw. AB - The contribution of calcitonin gene-related peptide (CGRP) to bilateral oedema formation in the rat hindpaw following an unilateral challenge with CGRP was investigated. Rats were injected into the left hindpaw with either saline, CGRP or a CGRP antagonist (CGRP8-37). All injections were given in a double blind fashion and in a volume of 100 microl. CGRP and CGRP8-37 were administered in concentrations of 75, 150 or 300 pmol. Volumes of the right and left hindpaw were measured every hour for 5 h by plethysmometry. Injection of CGRP 300 pmol into the left hindpaw resulted in a bilaterally increased hindpaw volume after 5 h as compared with the groups given saline. No changes were found in hindpaw volumes following the injection of either 75 or 150 pmol of CGRP or 75, 150 or 300 pmol of CGRP8-37 as compared with saline injection. To elucidate whether or not the bilateral oedema formation was related to a release of endogenous CGRP, microdialysis of the contralateral hindpaw was carried out, and concentrations of CGRP-like immunoreactivity (-LI) were determined by radioimmunoassay and high performance liquid chromatography. Injection of CGRP 300 pmol into the left hindpaw increased the release of CGRP-LI into the right hindpaw perfusate after 4 and 5 h. No changes in CGRP-LI were detected in the right hindpaw perfusate following challenge with saline or CGRP8-37. To study the contribution of the nervous system to the contralateral release of CGRP-LI, sciatic nerve ligated and intact sham-operated rats were used. Sciatic nerve ligation but not sham operation on the non-injected side abolished the increased release of CGRP-LI following contralateral administration of CGRP 300 pmol. To study the spinal cord mechanisms resulting in the bilateral oedema formation following unilateral challenge with 300 pmol of CGRP, intrathecal pretreatment with either 10 nmol bicuculline (GABA(A) receptor antagonist) or 10 nmol CGRP8-37 was carried out. Bicuculline but not CGRP8-37 abolished the bilateral oedema formation induced by CGRP 300 pmol. In order to study the mechanisms by which administration of CGRP 300 pmol induces oedema, CGRP 300 pmol was administered concomitantly with either 300 pmol of CGRP8-37 (CGRP receptor antagonist), or 3 nmol of promethazine (H1 receptor antagonist), or 3 nmol of s(-)-propranolol (5-HT1 receptor antagonist), or 3 nmol of cyproheptadine (5-HT2 receptor antagonist) or 3 nmol of ICS 205-930 (5-HT3 receptor antagonist). Oedema formation was measured at 1, 5, 7 and 24 h. Injection of CGRP 300 pmol into the left hindpaw induced a bilateral oedema formation which was still significant at 24 h. Concomitant administration of either CGRP8-37, ICS 205-920 or cyproheptadine blocked the oedema formation at 24 h. No effect on oedema formation was found when CGRP 300 pmol was co-administered with either promethazine or s(-)-propranolol (H1 and 5-HT1 receptor antagonists, respectively). The results of the present study show that both the nervous system and local inflammatory processes contribute to bilateral hindpaw oedema formation following unilateral challenge with CGRP 300 pmol. Our results indicate that endogenous release of CGRP following inflammatory response may play an important role in inducing oedema formation. PMID- 9863662 TI - Expression of cyclooxygenase-2 in foetal rat hepatocytes stimulated with lipopolysaccharide and pro-inflammatory cytokines. AB - Cyclooxygenase-2 (COX-2) is involved in the biosynthesis of prostanoids in the course of inflammatory reactions. This isoenzyme is regulated at the transcription level and many cells express COX-2 upon challenge with lipopolysaccharide (LPS) or pro-inflammatory cytokines. Since hepatocytes respond to LPS and pro-inflammatory stimuli, we investigated the expression of COX-2 in foetal and adult hepatocytes upon challenge with these substances. COX-2 was expressed in foetal hepatocytes incubated with LPS, tumour necrosis factor-alpha and interleukin-1beta. This response rapidly decreased after birth and was absent in hepatocytes from animals aged 2 days or more and treated under identical conditions. The expression of COX-2 was determined at the mRNA, protein and enzyme activity levels using Northern and Western blot, and following the synthesis of prostaglandin E2, respectively. The use of NS 398, a specific pharmacological inhibitor of COX-2, confirmed the expression of this isoenzyme in activated foetal hepatocytes. Synergism in COX-2 expression was observed between LPS, tumour necrosis factor-alpha and interleukin-1beta. Interleukin-6 and permeant analogues of cyclic AMP failed to induce COX-2 or to synergize with LPS. Also, transforming growth factor-beta inhibited the LPS- and pro-inflammatory cytokines-dependent expression of COX-2. These results indicate that foetal hepatocytes are competent to express COX-2 upon challenge with pro-inflammatory stimuli, a process lost completely in hepatocytes isolated from animals aged 2 days. PMID- 9863664 TI - Contractile responses elicited by hydrogen peroxide in aorta from normotensive and hypertensive rats. Endothelial modulation and mechanism involved. AB - The present study analyses the influence of hypertension and endothelium on the effect induced by hydrogen peroxide (H2O2) on basal tone in aortic segments from normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) of 6 month-old, as well as the possible mechanisms involved. Single (1 mM) or cumulative (100 nM-10 mM) concentrations of H2O2 produced a transient contraction or a concentration-dependent increase of basal tone, respectively, in segments from WKY and SHR. In both cases, the contractions were higher in intact segments from hypertensive than from normotensive rats, and increased by endothelium removal in both strains. Catalase (1000 u ml(-1), a H2O2 scavenger) abolished the contraction elicited by 1 mM H2O2 in both strains. Superoxide dismutase (SOD, 150 u ml(-1)) and dimethylsulphoxide (DMSO, 7 mM), scavengers of superoxide anions and hydroxyl radicals, respectively, did not alter H2O2-induced contractions in intact segments from both strains. However, L-NG-nitroarginine methyl ester (L NAME, 100 microM, a nitric oxide synthase inhibitor) increased the response to H2O2 in normotensive rats, although the increase was less than that produced by endothelium removal. Incubation of segments with 1 mM H2O2 for 15 min and subsequent washout reduced the contractile responses induced by 75 mM KCl in intact segments from SHR and in endothelium-denuded segments from both strains; this effect being prevented by catalase (1000 u ml(-1)). Indomethacin (10 microM, a cyclo-oxygenase inhibitor) and SQ 29,548 (10 microM, a prostaglandin H2/thromboxane A2 receptor antagonist) practically abolished the contractions elicited by H2O2 in normotensive and hypertensive rats. We conclude that: (1) the oxidant stress induced by H2O2 produces contractions mediated by generation of a product of the cyclo-oxygenase pathway, prostaglandin H2 or more probably thromboxane A2, in normotensive and hypertensive rats; (2) oxygen-derived free radicals are not involved in the effect of H2O2; (3) in normotensive rats, endothelium protects against H2O2-mediated injury to contractile machinery, determined by the impairment of KCl-induced contractions; and (4) endothelial nitric oxide has a protective role on the contractile effect induced by H2O2, that is lost in hypertension. PMID- 9863663 TI - Role of cyclo-oxygenase-2 induction in interleukin-1beta induced attenuation of cultured human airway smooth muscle cell cyclic AMP generation in response to isoprenaline. AB - Airway smooth muscle (ASM) in human asthma shows reduced relaxation and cyclic AMP generation in response to beta-adrenoceptor agonists. IL-beta attenuates cyclic AMP generation but the underlying mechanism is unclear. We have reported that IL-1beta induces cyclo-oxygenase-2 (COX-2) in human ASM cells and results in a marked increase in prostanoid generation with PGE2 and PGI2 as the major products. We investigated the role of COX-2 induction and prostanoid release (measured as PGE2) in IL-1beta induced attenuation of cyclic AMP generation in response to the beta-adrenoceptor agonist isoprenaline (ISO). Pre-treatment of human ASM cells with IL-1beta significantly attenuated cyclic AMP generation in response to high concentrations of ISO (1.0-10.0 microM) in a time- and concentration-dependent manner. The effect was accompanied by a high concentration of PGE2 release. The non-selective COX inhibitor indomethacin (Ind), the selective COX-2 inhibitor NS-398, the protein synthesis inhibitors cycloheximide (CHX) and actinomycin D and the steroid dexamethasone (Dex) all abolished the PGE2 release and prevented the attenuated cyclic AMP generation. COX substrate arachidonic acid time- and concentration-dependently mimicked IL 1beta induced attenuation and the effect was prevented by the non-selective COX inhibitors Ind and flurbiprofen, but not by NS-398, CHX and Dex. In contrast to IL-1beta, TNFalpha and IFNgamma, which are ineffective in inducing COX-2 and releasing PGE2 from human ASM cells, did not affect the cyclic AMP formation. Our study demonstrates that COX-2 induction and the consequent release of prostanoids plays a crucial role in IL-1beta induced attenuation of human ASM cell cyclic AMP response to ISO. PMID- 9863665 TI - Pharmacology of [3H]-pyrilamine binding and of the histamine-induced inositol phosphates generation, intracellular Ca2+ -mobilization and cytokine release from human corneal epithelial cells. AB - We recently reported on the successful generation of immortalized (CEPI-17-CL4) cells from primary human corneal epithelial (P-CEPI) cells which exhibited phenotypic, immunohistochemical and metabolic characteristics akin to the P-CEPI cells. The aims of the present studies were to investigate the ligand binding and functional coupling of the histamine receptors to various biochemical and physiological systems in the P-CEPI and CEPI-17-CL4 cells and to relate these findings to the normal and/or pathophysiological role of histamine on the human ocular surface. Specific [3H]-pyrilamine binding to CEPI-17-CL4 cell homogenates comprised >93% of the total binding and represented interaction with an apparent single population of high affinity (Kd=3.76+/-0.78 nM; n=4) and saturable (Bmax = 1582+/-161 fmol g(-1) tissue) number of histamine-1 (H1) receptor binding sites on CEPI-17-CL4 cell homogenates. The H1-receptor selective antagonists, pyrilamine (Ki=3.6+/-0.84 nM, n=4) and triprolidine (Ki = 7.7+/-2.6 nM, n=3), potently displaced [3H]-pyrilamine binding, while the H2- and H3-receptor selective antagonists, ranitidine and clobenpropit, were weak inhibitors (K(i)s>13 microM). Histamine induced phosphoinositide (PI) hydrolysis 2.7-4.4 fold above basal levels and with a potency of 14.9+/-4.9 microM (n=9) and 4.7+/ 0.2 microM (n=9) in P-CEPI and CEPI-17-CL4 cells, respectively. Histamine-induced PI turnover was antagonized by H1-receptor selective antagonist, triprolidine, with a potency (Ki) of 3.2+/-0.66 nM (n=10) and 3.03+/-0.8 nM (n=4) in P-CEPI and CEPI-17-CL4 cells, respectively, but weakly effected by 10 microM cimetidine and clobenpropit, H2- and H3-receptor antagonists. The PI turnover response was attenuated by pre-treatment of the cells with the selective phospholipase C inhibitor, U73122 (1-(6-((17beta-3-methoxyestra- 1,3,5(10)-trien-17 yl)amino)hexyl)-1H-pyrrole-2,5-dione) (IC50=4.8+/-2.4 microM, n = 3). Histamine stimulated intracellular Ca2+ ([Ca2+]i) mobilization in CEPI-17-CL4 cells with a potency of 6.3+/-1.5 microM (n=4). The histamine-induced [Ca2+]i mobilization was reduced by about 28% following pre-incubation of the cells with 4 mM EGTA. While triprolidine completely inhibited histamine-induced [Ca2+]i mobilization, it did not influence the bradykinin-induced [Ca2+]i mobilization response. Histamine (EC50s = 1.28-2.77 microM, n=3-4) concentration-dependently stimulated the release of interleukin-6 (IL-6), IL-8 and granulocyte macrophage colony stimulating factor, but it did not significantly alter release of tumour necrosis factor-alpha, PGE2 or collagenase-1 (matrix metalloproteinase-1; MMP-1) from CEPI cells. However, IL-1 (10 ng ml(-1)), foetal bovine serum (10%) and phorbol-12 myristate-13-acetate (3 microg ml(-1)) were effective positive control secretagogues of all the cytokines, PGE2 and MMP-1, respectively, from these cells. It is concluded that the CEPI cells express H1-histamine receptors which are positively coupled to PI turnover and [Ca2+]i mobilization which may be directly or indirectly responsible for the release of various cytokines from these cells at physiologically and/or pathologically relevant concentrations. PMID- 9863667 TI - Characterization of adenosine receptors evoking excitation of mesenteric afferents in the rat. AB - We examined the effects of adenosine receptor agonists and antagonists on the discharge of mesenteric afferent nerves supplying the jejunum in pentobarbitone sodium-anaesthetized rats. Adenosine (0.03-10 mg kg(-1), i.v.), NECA (0.3-300 microg kg(-1), i.v.) and the A1 receptor agonist, GR79236 (0.3-1000 microg kg( 1), i.v.), each induced dose-dependent increases in afferent nerve activity and intrajejunal pressure, hypotension and bradycardia. The A1 receptor antagonist, DPCPX (3 mg kg(-1), i.v.), antagonized all the effects of GR79236 but only the haemodynamic effects of adenosine and NECA. The A2A receptor antagonist, ZM241385 (3 mg kg(-1), i.v.), antagonized the hypotensive effect of NECA but none of the effects of GR79236. The A2A receptor agonist, CGS21680 (0.3-300 microg kg(-1), i.v.), and the A3 receptor agonist, IB-MECA (0.3-300 microg kg(-1), i.v.), each induced only a dose-dependent hypotension. Subsequent administration of adenosine (3 mg kg(-1), i.v.) induced increases in afferent nerve activity and intrajejunal pressure and bradycardia. ZM241385 (3 mg kg(-1), i.v.) antagonized the hypotensive effect of CGS21680 but not the effects of adenosine. Bethanechol (300 microg kg(-1), i.v.) evoked increases in afferent nerve activity and intrajejunal pressure, hypotension and bradycardia. However, adenosine (3 mg kg(-1), i.v.) evoked greater increases in afferent nerve activity than bethanechol despite inducing smaller increases in intrajejunal pressure. In summary, A1 and A2B and/or A2B-like receptors evoke adenosine-induced increases in mesenteric afferent nerve activity and intrajejunal pressure in the anaesthetized rat. Furthermore, elevations in intrajejunal pressure do not wholly account for adenosine-evoked excitation of mesenteric afferent nerves. PMID- 9863666 TI - Comparison of cannabinoid binding sites in guinea-pig forebrain and small intestine. AB - We have investigated the nature of cannabinoid receptors in guinea-pig small intestine by establishing whether this tissue contains cannabinoid receptors with similar binding properties to those of brain CB1 receptors. The cannabinoids used were the CB1-selective antagonist SR141716A, the CB2-selective antagonist SR144528, the novel cannabinoid receptor ligand, 6'-azidohex-2'-yne-delta8 tetrahydrocannabinol (O-1184), and the agonists CP55940, which binds equally well to CB1 and CB2 receptors, and WIN55212-2, which shows marginal CB2 selectivity. [3H]-CP55940 (1 nM) underwent extensive specific binding both to forebrain membranes (76.3%) and to membranes obtained by sucrose density gradient fractionation of homogenates of myenteric plexus-longitudinal muscle of guinea pig small intestine (65.2%). Its binding capacity (Bmax) was higher in forebrain (4281 fmol mg(-1)) than in intestinal membranes (2092 fmol mg(-1)). However, the corresponding KD values were not significantly different from each other (2.29 and 1.75 nM respectively). Nor did the Ki values for its displacement by CP55940, WIN55212-2, O-1184, SR141716A and SR144528 from forebrain membranes (0.87, 4.15, 2.85, 5.32 and 371.9 respectively) differ significantly from the corresponding Ki values determined in experiments with intestinal membranes (0.99, 5.03, 3.16, 4.95 and 361.5 nM respectively). The Bmax values of [3H]-CP55940 and [3H] SR141716A in forebrain membranes did not differ significantly from each other (4281 and 5658 fmol mg(-1)) but were both greater than the Bmax of [3H]-WIN55212 2 (2032 fmol mg(-1)). O-1184 (10 or 100 nM) produced parallel dextral shifts in the log concentration-response curves of WIN55212-2 and CP55940 for inhibition of electrically-evoked contractions of the myenteric plexus-longitudinal muscle preparation, its KD values being 0.20 nM (against WIN55212-2) and 0.89 nM (against CP55940). We conclude that cannabinoid binding sites in guinea-pig small intestine closely resemble CB1 binding sites of guinea-pig brain and that 0-1184 behaves as a cannabinoid receptor antagonist in the guinea-pig myenteric plexus longitudinal muscle preparation. PMID- 9863668 TI - A microdialysis study of the in vivo release of 5-HT in the median raphe nucleus of the rat. AB - The present study has examined several characteristics of the release of 5-HT in the median raphe nucleus in terms of its dependence of nerve impulse, provenance of a vesicular storage fraction as well as the regulatory role played by 5-HT1A receptors. Tetrodotoxin (1 microM) and reserpine (5 mg kg(-1), i.p.) virtually suppressed the output of 5-HT. The administration of EEDQ (10 mg kg(-1), i.p.) did not alter the basal release of 5-HT but abolished the reduction of 5-HT release induced by 8-OH-DPAT (0.1 mg kg(-1), s.c.). The perfusion of 1-100 microM of 8-OH-DPAT or the novel 5-HT1A agonist BAY x 3702 decreased the efflux of 5-HT, whereas the perfusion of the 5-HT1A antagonist WAY-100635 failed to alter 5-HT release. The decrease in dialysate 5-HT induced by 100 microM 8-OH-DPAT was reversed by the concurrent perfusion of 100 microM WAY-100635. Also, the perfusion of 100 microM WAY-100635 for 2 h inhibited partly the reduction of 5-HT release evoked by the systemic administration of 8-OH-DPAT (0.1 mg kg(-1)). These results indicate that extracellular 5-HT in the median raphe nucleus is stored in vesicles and released in an impulse-dependent manner. Also, the basal release of 5-HT in the median raphe nucleus does not appear to be under the tonic control of somatodendritic 5-HT1A receptors by endogenous 5-HT. Instead, this feedback mechanism seems to be triggered when an excess of the transmitter or a 5-HT1A agonist is present in the extracellular space of the median raphe nucleus. PMID- 9863669 TI - Evidence for two different types of P2 receptors stimulating insulin secretion from pancreatic B cell. AB - Adenine nucleotides have been shown to stimulate insulin secretion by acting on P2 receptors of the P2Y type. Since there have been some discrepancies in the insulin response of different analogues of ATP and ADP, we investigated whether two different types of P2 receptors exist on pancreatic B cells. The effects of alpha,beta-methylene ATP, which is more specific for the P2X subtype, were studied in vitro in pancreatic islets and isolated perfused pancreas from rats, in comparison with the potent P2Y receptor agonist ADPbetaS. In isolated islets, incubated with a slightly stimulating glucose concentration (8.3 mM), alpha,beta me ATP (200 microM) and ADPbetaS (50 microM) similarly stimulated insulin secretion; by contrast, under a non stimulating glucose concentration (3 mM), alpha,beta-me ATP was still effective whereas ADPbetaS was not. In the same way, in islets perifused with 3 mM glucose, alpha,beta-me ATP but not ADPbetaS induced a partial but significant reduction in the peak 86Rb efflux induced by the ATP dependent potassium channel opener diazoxide. In the isolated pancreas, perfused with a non stimulating glucose concentration (4.2 mM), ADPbetaS and alpha,beta-me ATP (5-50 microM), administered for 10 min, induced an immediate, transient and concentration-dependent increase in the insulin secretion; their relative potency was not significantly different. In contrast, with a slightly stimulating glucose concentration (8.3 mM), ADPbetaS was previously shown to be 100 fold more potent than alpha,beta-me ATP. Furthermore, at 4.2 mM glucose a second administration of alpha,beta-me ATP was ineffective. In the same way, ADPbetaS was also able to desensitize its own insulin response. At 3 mM glucose, alpha,beta-me ATP as well as ADPbetaS (50 microM) induced a transient stimulation of insulin secretion and down regulated the action of each other. These results give evidence that pancreatic B cells, in addition to P2Y receptors, which potentiate glucose induced insulin secretion, are provided with P2X receptors, which transiently stimulate insulin release at low non-stimulating glucose concentration and slightly affect the potassium conductance of the membrane. PMID- 9863670 TI - Modulation of inhibitory post-synaptic currents (IPSCs) in mouse cerebellar Purkinje and basket cells by snake and scorpion toxin K+ channel blockers. AB - Using an in vitro mouse cerebellar slice preparation and whole-cell electrophysiological recording techniques we have characterized Purkinje and basket cell inhibitory post-synaptic currents (IPSCs), and examined the effects of a number of selective peptidergic K+ channel blockers. Spontaneous IPSC amplitude ranged from approximately 10 pA up to approximately 3 nA for both cell types [mean values: Purkinje cells -122.8+/-20.0 pA (n = 24 cells); basket cells 154.8+/-15.9 pA (n = 26 cells)]. Frequency varied from approximately 3 up to approximately 40 Hz, [mean values: basket cells 14.9+/-1.7 Hz (n=26 cells); Purkinje cells 17.9+/-2.2 Hz (n=24 cells)]. 5 microM bicuculline eliminated virtually all spontaneous currents. IPSC rise times were fast (approximately 0.6 ms) and the decay phase was best fit with the sum of two exponential functions (tau1 and tau2: approximately 4 ms and approximately 20 ms, n=40; for both cell types). The snake toxins alpha-dendrotoxin (alpha-DTX) and toxin K greatly enhanced IPSC frequency and amplitude in both cell types; the closely related homologues toxin I and gamma-dendrotoxin (gamma-DTX) produced only marginal enhancements (all at 200 nM). Two scorpion toxins, margatoxin (MgTX) and agitoxin 2 (AgTX-2) had only minor effects on IPSC frequency or amplitude (both at 10 nM). Low concentrations of tetraethylammonium (TEA; 200 microM) had no overall effect on cerebellar IPSCs, whilst higher concentrations (10 mM) increased both the frequency and amplitude. The results suggest that native K+ channels, containing Kv1.1 and Kv1.2 channel subunits, play an influential role in controlling GABAergic inhibitory transmission from cerebellar basket cells. PMID- 9863672 TI - Human perspiration measurement. AB - We review various methods developed for human perspiration measurement and their physiological applications, with special reference to the performance and application of a new home-made ratemeter and instrumentation with a microscope. Many kinds of humidity sensor based on humidity-sensitive electrical properties have been investigated and placed on the market. Recently a capacitive thin-film humidity sensor was constructed and confirmed to be one of the best humidity sensors for accurately and quickly detecting changes in the relative humidity of gas-flow perfused through a ventilated chamber for human perspiration measurement. In this paper we also introduce a new home-made ratemeter with a capacitive humidity sensor, the electrical output of which is not disturbed by changes in ambient temperature, and new instrumentation for directly observing drops of sweat secreted from eccrine glands in human skin and simultaneously measuring the change in amount of perspiration at the same area of skin. Finally, we review physiological applications of the methods for measuring human palmar perspiration including emotional sweating. PMID- 9863671 TI - Insensitivity of volume-sensitive chloride currents to chromones in human airway epithelial cells. AB - Chromones (sodium cromoglycate and sodium nedocromil) block cell swelling activated Cl- channels in NIH-3T3 fibroblasts and endothelial cells. This has led to hypothesize that cell volume regulation might be involved in asthma pathogenesis. Using whole-cell patch-clamp experiments, we studied the effect of chromones on volume-sensitive Cl- currents in transformed human tracheal epithelial cells (9HTEo-) and in primary cultures of human bronchial epithelial cells (BE). Cl- currents activated by hypotonic shock were poorly blocked by extracellular nedocromil or cromoglycate. The block was voltage-dependent since it was observed only at positive membrane potentials. At the concentration of 5 mM, the current inhibition by both chromones at +80 mV was about 40% for 9HTEo- and only 20% for BE. Intracellular application of chromones elicited a voltage independent inhibition in 9HTEo- cells. Under this condition, volume-sensitive Cl currents were reduced at all membrane potentials (60 and 45% inhibition by 2 mM nedocromil and cromoglycate respectively). In contrast intracellular chromones were ineffective in BE cells. The relative refractoriness to chromones, in contrast with the high sensitivity shown by other Cl- channels, suggests that the epithelial volume-sensitive Cl- channel is not involved in asthma. PMID- 9863673 TI - An investigation into the spatial relationship between complexity and motility within the oesophagus. AB - Nonlinear analysis techniques have recently been used in the characterization of complex physiological signals seen in pathological disorders such as epilepsy and cardiac fibrillation. In this study a series of controlled swallows from an asymptomatic demonstration group was investigated using oesophageal manometry. The nonlinear measure of complexity, largest Lyapunov exponents and phase portraits were then used to explore the complexity of motility patterns at different points within the oesophagus. Results indicate greater complexity within the region of the striated muscle in the upper oesophagus than that observed within the region of smooth muscle in the lower oesophagus. Phase portraits showed that manometry patterns within the asymptomatic demonstration group could be quite different, highlighting the problems in clinical diagnosis. The characterization of motility disorders associated with complex manometry patterns such as diffuse oesophageal spasm (DOS) and nonspecific motility disorder (NOMD) still represents a diagnostic challenge. The use of nonlinear techniques enabling the quantitative and qualitative measurement of oesophageal complexity is considered in the classification of such disorders. PMID- 9863674 TI - Ventilation and perfusion imaging by electrical impedance tomography: a comparison with radionuclide scanning. AB - Electrical impedance tomography (EIT) is a technique that makes it possible to measure ventilation and pulmonary perfusion in a volume that approximates to a 2D plane. The possibility of using EIT for measuring the left-right division of ventilation and perfusion was compared with that of radionuclide imaging. Following routine ventilation (81mKr) and perfusion scanning (99mTc-MAA), EIT measurements were performed at the third and the sixth intercostal level in 14 patients with lung cancer. A correlation (r = 0.98, p < 0.005) between the left right division for the ventilation measured with EIT and that with 81mKr was found. For the left-right division of pulmonary perfusion a correlation of 0.95 (p < 0.005) was found between the two methods. The reliability coefficient (RC) was calculated for estimating the left-right division with EIT. The RC for the ventilation measurements was 94% and 96% for the perfusion measurements. The correlation analysis for reproducibility of the EIT measurements was 0.95 (p < 0.001) for the ventilation and 0.93 (p < 0.001) for the perfusion measurements. In conclusion, EIT can be regarded as a promising technique to estimate the left right division of pulmonary perfusion and ventilation. PMID- 9863675 TI - The influence of extravascular lung water on cardiac output measurements using thoracic impedance cardiography. AB - The purpose of this study was to investigate the influence of pulmonary oedema as measured with the double indicator dilution technique on the accuracy of cardiac output (CO) measurement using thoracic impedance cardiography (TIC) compared with thermodilution in thirteen sepsis patients. Differences in the Kubicek and Sramek Bernstein equation with respect to pulmonary oedema were explored theoretically and experimentally. From a parallel two cylinder model a hypothesis can be derived that CO determined with the Kubicek equation is oedema independent, whereas CO determined using the Sramek-Bernstein equation is oedema dependent. Experimentally, CO determined using Kubicek's equation correlated better with thermodilution CO (r = 0.75) than CO determined with the Sramek-Bernstein equation (r = 0.25). The effect of oedema on the accuracy of TIC was investigated by comparing the differences in the CO of impedance and thermodilution to the extravascular lung water index. For the Kubicek equation the difference was not influenced by oedema (r = 0.04, p = 0.84), whereas for the Sramek-Bernstein equation the difference was affected by oedema (r = 0.39, p = 0.05). Thus, the effects of pulmonary oedema on the accuracy of TIC measurements can better be understood with the parallel cylinder model. Moreover, the Kubicek equation still holds when pulmonary oedema is present, in contrast to the Sramek-Bernstein equation. PMID- 9863676 TI - Quantification of blood volume by electrical impedance tomography using a tissue equivalent phantom. AB - An in vivo electrical impedance tomography (EIT) system was designed to accurately estimate quantities of intra-peritoneal blood in the abdominal cavity. For this it is essential that the response is relatively independent of the position of the high conductivity anomaly (blood) in the body. The sensitivity of the system to the introduction of blood-equivalent resistivity anomalies was assessed by using a cylindrical tissue-equivalent phantom. It was found that a satisfactorily uniform response of the system in both radial (transverse) and axial (longitudinal) directions in the phantom could be achieved by filtering resistivity profile images obtained by EIT measurement, and by using extended electrodes to collect data. Post-processing of single impedance images gave rise to a quantity denoted the resistivity index. A filter was then used to remove the remaining radial variation of the resistivity index. It was calculated by evaluating the resistivity index of a number of theoretically calculated images, and constructing a correction filter similar to those used to remove lens imperfections, such as coma, in optical components. The 30% increase in the resistivity index observed when an anomaly was moved to the maximum extent allowed by the filter calculation (0.75 of the phantom radius) was reduced by the filter to 6%. A study of the axial dependence observed in the resistivity index using electrodes extended in the axial direction by +/-5 cm found that the variation in resistivity index with axial position was about half of that observed using small circular electrodes similar to those used in the Sheffield mark I system. PMID- 9863677 TI - Extra-cellular volume estimation by electrical impedance--phase measurement or curve fitting: a comparative study. AB - In order to determine body fluid shifts between the intra- and extra-cellular spaces, multifrequency impedance measurement is performed. According to the Cole Cole extrapolation, lumped values of intra- and extra-cellular conduction can be estimated which are commonly expressed in resistances Ri and Re respectively. For this purpose the magnitude and phase of the impedance under study are determined at a number of frequencies in the range between 5 kHz and 1 MHz. An approach to determine intra- and extra-cellular conduction on the basis of Bode analysis is presented in this article. On this basis, estimation of the ratio between intra- and extra-cellular conduction could be performed by phase measurement only, midrange in the bandwidth of interest. An important feature is that the relation between intra- and extra-cellular conduction can be continuously monitored by phase measurement and no curve fitting whatsoever is required. Based on a two frequency measurement determining Re at 4 kHz and phi(max) at 64 kHz it proved possible to estimate extra-cellular volume (ECV) more accurately compared with the estimation based on extrapolation according to the Cole-Cole model in 26 patients. Reference values of ECV were determined by sodium bromide. The results show a correlation of 0.90 with the reference method. The average error of ECV estimation was -3.6% (SD 8.4), whereas the Cole-Cole extrapolation showed an error of 13.2% (SD 9.5). An important feature of the proposed approach is that the relation between intra- and extra-cellular conduction can be continuously monitored by phase measurement and no curve fitting whatsoever is required. PMID- 9863678 TI - Pelvic bioelectrical impedance measurements to detect rectal filling. AB - Anorectal sensory deficits are an important cause of defecatory disorders and are also a reason for evacuatory difficulties in patients undergoing total anorectal reconstruction. A method to detect rectal filling would be beneficial in such patients. We have investigated the feasibility of detecting rectal filling in vitro and in vivo by measuring changes in pelvic impedance. In vitro, a model of the pelvis was constructed using a cylindrical plastic tank filled with an electrolyte solution (conductivity 3 mS cm(-1)). Conductive Visking tubing representing the rectum was suspended in the tank and incrementally filled with artificial faeces. An impedance meter detected changes in voltage on rectal filling when an alternating current of 2 mA was passed at eight frequencies (4.8 to 612 kHz). In vivo, changes in pelvic bioelectrical impedance upon retrograde and antegrade rectal filling with artificial faeces were evaluated in three pigs, four electrodes being implanted in the pelvis. Impedance measurements accurately detected 'rectal' volume changes in vitro (n = 10, p < 0.0001; Kruskal Wallis), but not in vivo (n = 68, p = 0.48; Kruskal Wallis). This was probably due to extreme sensitivity of the detecting device to movement, a problem that needs to be resolved before this technique could be used in man. PMID- 9863679 TI - Measurement of the volume of a leg ulcer using a laser scanner. AB - Effective management of leg ulcer healing depends on accurate and reliable measurements of wound size. Current techniques usually rely on estimates of surface area or circumference which do not fully describe the healing process. A novel instrument has been developed that is capable of measuring the variations in the surface contours (topography) of a solid object. Its primary application is to measure the size of a leg ulcer by scanning a laser displacement sensor over the affected area. There is no contact with the wound and scanning takes approximately 2 minutes to perform. Volume is calculated by subtracting the measured topography from one calculated using an algorithm to reconstruct a healthy leg surface. A study was carried out where patients had their ulcers scanned during their visits to the leg ulcer clinic. Data are presented from two venous leg ulcers showing the calculated volume reducing over time. PMID- 9863680 TI - Lateralized use of the mouth in production of vocalizations by marmosets. AB - We have found that the common marmoset, Callithrix jacchus, displays a larger left hemimouth during production of fear expressions, with or without vocalization, and a larger right hemimouth when producing a social contact call. Thus, marmosets have right hemisphere specialization for the production of negative emotional expressions and vocalizations and left hemisphere specialization for the production of social contact communication. These hemispheric specializations for social communication in marmosets are the same as those found in humans for speech production and for the control of emotional expressions. We suggest that hemispheric specializations for communication in humans may well have precursors in primate evolution. PMID- 9863681 TI - Relative hand skill predicts academic ability: global deficits at the point of hemispheric indecision. AB - Population variation in handedness (a correlate of cerebral dominance for language) is in part genetic and, it has been suggested, its persistence represents a balanced polymorphism with respect to cognitive ability. This hypothesis was tested in a sample of 12,770 individuals in a UK national cohort (the National Child Development Study) by assessing relative hand skill (in a square checking task) as a predictor of verbal, non-verbal, and mathematical ability and reading comprehension at the age of 11 years. Whereas some modest decrements were present in extreme right handers the most substantial deficits in ability were seen close to the point of equal hand skill ('hemispheric indecision'). For verbal ability females performed better than males, but the relationship to relative hand skill was closely similar for the two sexes; for reading comprehension males close to the point of equal hand skill showed greater impairments than females. Analysed by writing hand the relationship of ability to hand skill appeared symmetrical about the point of 'hemispheric indecision'. The variation associated with degrees of dominance may reflect the operation of continuing selection on the gene (postulated to be X-Y linked) by which language evolved and speciation occurred. PMID- 9863682 TI - Prefrontal involvement in "temporal bridging" and timing movement. AB - Brain activity exclusively related to a temporal delay has rarely been investigated using modern brain imaging. In this study we exploited the temporal resolution of functional magnetic resonance imaging (fMRI) to characterise, by sinusoidal regression analysis, differential neuroactivation patterns induced in healthy subjects by two sensorimotor synchronization tasks different in their premovement delay of either 0.6 s or 5 s. The short event rate condition required rhythmic tapping, while the long event rate condition required timing of intermittent movements. Left rostral prefrontal cortex, medial frontal cortex, SMA and supramarginal gyrus demonstrated increased MR signal intensity during low frequency synchronization, suggesting that these brain regions form a distributed neural network for cognitive time management processes, such as time estimation and motor output timing. Medial frontal cortex showed a biphasic pattern of response during both synchronization conditions, presumably reflecting frequency independent motor output related attention. As predicted, sensorimotor and visual association areas demonstrated increased MR signal intensity during high frequency synchronization. PMID- 9863683 TI - Identification without manipulation: a study of the relations between object use and semantic memory. AB - The role of semantic knowledge in object utilisation is a matter of debate. It is usually presumed that access to semantic knowledge is a necessary condition for manipulation, but a few reports challenged this view. The existence of a direct, pre-semantic route from vision to action has been proposed. We report the case of a patient with a disorder of object use in everyday life, in the context of probable Alzheimer's disease. This patient was also impaired when manipulating single objects. He showed a striking dissociation between impairment in object use and preserved capacity to perform symbolic and meaningless gestures. To elucidate the nature of the disorder, and to clarify the relations between semantic knowledge and object use, we systematically assessed his capacity to recognise, name, access semantic knowledge, and use 15 common objects. We found no general semantic impairment for the objects that were not correctly manipulated, and, more importantly, no difference between the semantic knowledge of objects correctly manipulated and objects incorrectly manipulated. These data, although not incompatible with the hypothesis of a direct route for action, are better accommodated by the idea of a distributed semantic memory, where different types of knowledge are represented, as proposed by Allport (Allport, D. A. Current perspectives in dysphasia, pp. 32-60. Churchill Livingstone, Edinburgh, 1985). PMID- 9863684 TI - Linguistic and non-linguistic abilities in a patient with early left hemispherectomy. AB - A patient who underwent early removal of the left hemisphere because of Sturge Weber syndrome was submitted to detailed linguistic and visuospatial batteries. The performances were compared to performances of subjects matched for age, education and IQ, but without focal cerebral lesions. Language was mildly impaired but to the same extent as in IQ controls. On the contrary, visuospatial abilities were clearly worse than in IQ controls, and the most preserved visuospatial abilities seemed to be the less sophisticated ones. Non literal comprehension of language, a function generally attributed to the right hemisphere, was intact. This same pattern, that is, preservation of language and impairment of visuospatial abilities, also seems to occur in subjects who have undergone surgical removal of the right hemisphere; in other words, the cognitive pattern seems the same regardless of which hemisphere is removed. These observations suggest that no matter which hemisphere is removed, functional reorganisation follows a hierarchical criterion which privileges the linguistic function, and the visuospatial functions most essential for independent survival. PMID- 9863685 TI - Going, going, gone...? Implicit and explicit test of conceptual knowledge in a longitudinal study of semantic dementia. AB - Patients suffering from semantic dementia provide important constraints on theories of the structure and organisation of semantic memory. In this article we report one such patient, AM, whose progressive deterioration of semantics enables us to address the much-debated issue of whether conceptual structure is hierarchically organised. The hierarchical account predicts that brain damage should impair lower levels of the hierarchy (property information) before affecting higher level (category) information (Warrington and Shallice, Q. J. Exp. Psychol. 1975, 27, 635-657). We evaluate this prediction by repeated testing of AM in two studies--a semantic priming task and a verification task--over an 18 month period, contrasting the progressive deterioration of properties (functional and perceptual) and category relations (category co-ordinates and category labels). Properties were preserved longer than category information, arguing against a hierarchical account of semantic memory. In addition, functional properties were most robust to brain damage, supporting our claim that functional information plays a special role in semantic representations (Durrant-Peatfield et al., Proc. 19th Ann. Conf. of the Cognitive Science Society. Erlbaum, Mahwah, NJ, 1997. pp. 193-198. Tyler et al., Cognitive Neuropsychol. 1997, 14, 511-545). PMID- 9863686 TI - Monitoring for target objects: activation of right frontal and parietal cortices with increasing time on task. AB - The right prefrontal and parietal cortices have been implicated in attentional processing in both neuropsychological and functional neuroimaging literature. However, attention is a heterogeneous collection of processes, each of which may be underpinned by different neural networks. These attentional networks may interact, such that engaging one type of attentional process could influence the efficiency of another via overlapping neural substrates. We investigated the hypothesis that right frontal and parietal cortices provide the neuroanatomical location of the functional interaction between sustained attention and the process of selectively monitoring for target objects. Six healthy volunteers performed one of two tasks which required either selective or non-selective responding. The task lasted continuously for 18 min, during which time 3 Positron Emission Tomography (PET) scans were acquired for each task. This was repeated to obtain 12 PET measurements of regional cerebral blood flow (rCBF) for each subject. The right inferior frontal and parietal cortices were differentially activated by increasing time on task during the selective (S) vs non-selective (NS) task. Specifically, rCBF decreased with increasing time spent performing the NS task but not the S task. This result suggests that the normal deactivation in these areas as time on task increases is counteracted by the extra cognitive demands of selectively responding to target objects. Therefore, we have confirmed our hypothesis that right frontal and parietal cortices provide the neuroanatomical location for the modulation of object selection by sustained attention. We also identified the neuroanatomical correlates of each process separately, and confirmed earlier reports of prefrontal cortex and anterior cingulate activation associated with selective responding, and a fronto-parietal thalamic network associated with sustained attention. PMID- 9863687 TI - Focal retrograde amnesia associated with vascular headache. AB - We report the case of a 42-year-old man with repeated attacks of headache associated with retrograde amnesia. Neuropsychological tests before and after the major episode of amnesia showed mild neuropsychological deficits but with spared anterograde memory and learning functions. The amnesia was dense for a period of 15-20 years and included people and events (public and private). There was also a suggestion of amnesia for learned skills. Neurologically he had mild clinical signs and focal EEG-abnormalities in the left fronto-temporal region, but CT, MRI, and SPECT showed no abnormality. Five years after the onset of amnesia there was no recovery of the retrograde memory deficit, but a PET (glucose) scan was normal and neuropsychological testing showed no deficits. An association with migraine has been reported for some non-classical amnesias, but this is the first case of selective retrograde amnesia in a patient with headache as a primary neurological diagnosis. PMID- 9863688 TI - Baseline EEG asymmetries and performance on neuropsychological tasks. AB - Thirty-two participants were tested for both resting electroencephalography (EEG) and neuropsychological function. Eight one-minute trials of resting EEG were recorded from 14 channels referenced to linked ears, which was rederived to an average reference. Neuropsychological tasks included Verbal Fluency, the Tower of London, and Corsi's Recurring Blocks. Asymmetries in EEG alpha activity were correlated with performance on these tasks. Similar patterns were obtained for delta and theta bands. Factor analyses of resting EEG asymmetries over particular regions suggested that asymmetries over anterior scalp regions may be partly independent from those over posterior scalp regions. These results support the notions that resting EEG asymmetries are specified by multiple mechanisms along the rostral/caudal plane, and that these asymmetries predict task performance in a manner consistent with lesion and neuroimaging studies. PMID- 9863690 TI - Directed attention after unilateral frontal excisions in humans. AB - We investigated the ability of patients with frontal-lobe lesions to benefit from advance information in a simple reaction-time task. The task involved pressing a button in response to the appearance of a peripheral target (visual angle of 11.5 degrees). A cue, presented in the centre of the screen, preceded the target onset by either a short (average 500 ms) or a long (average 3000 ms) interval. In half of the trials, the cue was an arrow indicating the location, in the left or right hemifield, of the upcoming target; in the other half, the cue was an uninformative plus sign. In addition to patients with unilateral excisions of frontal cortex, we tested patients with anterior temporal-lobe excisions and normal controls. The frontal-lobe group was mildly impaired with respect to the temporal-lobe group in using advance spatial cues to speed response to the visual target. While the size of the cueing effect changed across the range of cue target intervals tested, there was no variation across intervals in the size of the impairment exhibited by the frontal-lobe group. The site and the volume of the lesions were determined on the basis of magnetic resonance image (MRI) scans in 10 of the 17 patients in the frontal group. There was no correlation between lesion volume and benefit score in these patients. PMID- 9863689 TI - Executive processes in Parkinson's disease--random number generation and response suppression. AB - In producing random numbers, subjects typically deviate systematically from statistical randomness. It is considered that these biases reflect constraints imposed by underlying structures and processes, rather than a deficient concept of randomness. Random number generation (RNG) places considerable demands on executive processes, and provides a possibly useful tool for their investigation. A group of patients with Parkinson's disease (PD) and a group of controls were tested on a RNG task, both alone and with a concurrent attention-demanding task (manual tracking). Both groups showed the biases in RNG described previously, including a strong counting tendency and repetition avoidance. Overall RNG performance did not differ between the groups, although differences were found in the counting biases in the patient and control groups, with the controls showing a bias towards counting in twos, and the patients a bias towards counting in ones. The secondary task reversed the bias shown by controls and exacerbated the bias in the patients. A network modulation model may help explain many of the features of RNG. We suggest that naturally biased output from an associative network must be actively suppressed by an attention-demanding, limited-capacity process. This suppression may be disrupted by the pathophysiology of PD and by concurrent tasks. Convergent evidence from various sources is discussed which supports a role of the dorsolateral prefrontal cortex (DLPFC) in this process. PMID- 9863691 TI - Hemispheric specialization in the detection of subjective objects. AB - Three experiments were conducted to examine hemispheric specialization for the detection of subjective objects. In the first two experiments, observers searched for the presence of a square defined by subjective contours. The first experiment demonstrated that the left hemisphere made more errors for detecting these objects. The second experiment showed that the increased errors were due to the left hemisphere responding to the individual features of the objects and not the objects as a whole. In the second experiment, the right hemisphere was also faster for detecting the absence of a subjective object. A third experiment was conducted to determine if performance for the right hemisphere was due to object level processing. It was shown that the right hemisphere only makes illusory conjunctions for features within perceptual groups while the left hemisphere makes illusory conjunctions both within and across perceptual groups, providing converging evidence for object level processing in the right hemisphere. The results suggest that the right hemisphere conjoins feature information for the perception of objects. PMID- 9863692 TI - Frequency judgements of abstract designs by patients with diencephalic, temporal lobe or frontal lobe lesions. AB - Patients with diencephalic, temporal lobe or frontal lobe lesions were compared with healthy controls on a frequency judgement task. The three patient groups were disproportionately impaired at estimating how often a series of abstract designs had been presented relative to controls. Diencephalic and temporal lobe patients did not differ from each other. It is argued that the results may reflect a 'core' memory deficit in the temporal lobe patients. The impairment in the frontal patients may reflect their difficulty in making an organised search in memory for multiple traces of an item, while the deficit shown by the diencephalic patients (particularly those with Korsakoff syndrome) may be due to the combined effects of a generally poor memory and superimposed frontal pathology. PMID- 9863693 TI - Sex differences in oral asymmetries during wordrepetition. AB - During speech production the right side of the mouth is opened to a larger degree in most people. This facial asymmetry is thought to be related to a left hemisphere dominance in language processing and/or motor programming. We investigated asymmetrical lip separations during discrete or serial word productions in right handed persons. The results revealed a right sided lip separation bias in both genders during discrete word production in which the words had to be uttered once. As soon as the words had to be produced continuously, however, a clear sex difference appeared with males having the usual right bias but females now showing no clear asymmetry, with a tendency for larger lip separations on the left side. These results suggest the existence of two separate neural systems from which one controls the discrete task and which is left hemisphere dominant in both genders. The other is probably involved in serial word productions and shows a sex difference with regard to its asymmetry pattern. PMID- 9863694 TI - Word repetition within- and across-visual fields: an event-related potential study. AB - A divided visual field (DVF) procedure was used to investigate the scalp distribution of the event-related potential (ERP) repetition effect. ERPs were recorded from 27 scalp sites whilst subjects (n = 20) discriminated between words and non-words presented to either the left (LVF) or the right (RVF) visual field. A proportion of the words were repeated on the trial immediately following their first presentation. In two within-field repetition conditions the two encounters with a word occurred in the same visual field (LVF or RVF). In two across-field repetition conditions, the two encounters with a word occurred in different visual fields. For both words and non-words, task performance was better for RVF presentations than for LVF presentations. In each repetition condition there was a positive-going shift in the ERP elicited by repeated words compared to that elicited by words on their first presentation. This ERP repetition effect was equivalent in magnitude and lateralised to the right hemisphere to an equivalent degree in all four repetition conditions. It is suggested that the ERP effects largely reflect the processing of visual form thought to occur predominately in the right hemisphere. PMID- 9863695 TI - The effects of aging on layer 1 in area 46 of prefrontal cortex in the rhesus monkey. AB - The effect of age on layer 1 of area 46 of prefrontal cortex was determined in the cerebral cortices of 15 rhesus monkeys, 13 of which had been behaviorally tested. Five of the monkeys were young (5-7 years of age), three were middle-aged (9-12 years) and seven were old (24-32 years). It was found that with age, layer 1 becomes significantly thinner and the glial limiting membrane becomes thicker. Counts of synapses in layer 1 of seven of these monkeys using the physical disector method on thin sections revealed that compared to young monkeys, there is a 30-60% reduction in the density of synapses per unit volume in old monkeys. This loss of synapses is accompanied by a reduction in the frequency of profiles of postsynaptic dendrites and their spines from the neuropil of layer 1, indicating that some spiny dendrites that belong to the apical dendritic tufts of pyramidal cells are degenerating and being lost with age. Correlation of these morphological changes with the behavioral data shows that there is a significant correlation between the thickness of layer 1 and memory function, as measured by the 2 min delay condition of the delayed non-matching to sample task. Also, there is significant correlation between the numerical density of synapses in layer 1 and three of the behavioral measures used, as well as the Cognitive Impairment Index. Thus, the changes that occur with age in layer 1 provide one possible basis for the age-related cognitive impairment evidenced in monkeys and humans alike. PMID- 9863696 TI - Area V4 in Cebus monkey: extent and visuotopic organization. AB - We used electrophysiological mapping and myeloarchitectural criteria in order to define the location, extent and visual topography of the fourth visual area (V4) in anesthetized and paralyzed Cebus monkey. Based on these criteria, the borders of V4 with surrounding areas were defined both on the dorsal and ventral cortical surfaces. In addition, to better visualize the visuotopic organization and to evaluate its regularity, we constructed bidimensional maps and projected the recording sites onto them. Area V4 has an almost complete representation of the binocular visual field with the lower visual field represented dorsally (V4d) and the upper field ventrally (V4v). We found this representation to be more extensive than those previously described. The representation of the central portion of the visual field is largely expanded in comparison with that of the periphery. This emphasis in central vision could be related with the involvement of V4 in the ventral stream of visual information processing. Receptive field size increases with increasing eccentricity, while cortical magnification factor decreases. The cortical magnification factor measured along isopolar lines is, on average, 1.5-2.0 times greater than that measured along the isoeccentric lines, suggesting the existence of a small anisotropy in central and peripheral V4. PMID- 9863697 TI - Glutamatergic control of the expression of the proenkephalin gene in rat frontoparietal cortical slice cultures. AB - The expression of the proenkephalin (PEnk) gene in rat neocortex develops during the first two postnatal weeks in an outside first-inside last mode that is opposite to the gradient of neurogenesis. To test whether the distribution of PEnk gene expression depends on the formation of the local circuitry, we examined the role of glutamate neurons in the expression of the gene in slice cultures of rat frontoparietal cortex. In situ and Northern blot hybridization were used for analysis. In slices explanted at postnatal day 6, the neuronal expression of the PEnk gene developed as in vivo. The expression responded to glutamate receptor agonists and antagonists in a time-dependent manner. After 2 days in vitro the expression of the gene was only enhanced by N-methyl-D-aspartate receptors, whereas after 7 days in vitro AMPA receptors also regulated the expression. We concluded that glutamate neurons are involved in the development and maintenance of the PEnk gene expression in the neocortex. PMID- 9863698 TI - Variation in hippocampal neuron number with age and brain volume. AB - Hippocampal size and neuron number are reduced in a number of conditions, including temporal lobe epilepsy and Alzheimer's disease. Furthermore, a decrease with advancing age has also been suggested. The present study examined the entire hippocampal formation of 12 subjects aged from 46 to 85 years and free from neurological disease. The volume of seven subregions (CA1, CA2-3, CA4, dentate gyrus, subiculum, presubiculum and white matter) was determined and the number of neurons estimated in each of these grey matter subregions using the optical dissector technique. There was a significant relationship between CA1 neuron number and cerebrum volume. Multivariate analysis showed the greater contribution to the variance in CA1 neuron number was made by cerebrum volume (69%) rather than age (2%) or sex (1%). The findings of this study show that, in neurologically normal individuals, brain size is a major determinant of the number of CA1 neurons. PMID- 9863699 TI - Long-term potentiation in the neocortex of the adult, freely moving rat. AB - Neocortical preparations have proven highly resistant to the induction of long term potentiation (LTP), and we have only recently determined the conditions sufficient for the induction of neocortical LTP in the adult, freely moving rat. The stimulation trains must be spaced and repeated over a period of days in order to reach asymptotic levels of potentiation. Here we show that, within these constraints, the neocortex is actually highly responsive. LTP could be induced with as few as one brief high frequency train per day or with extremely low intensity stimulation trains. We also provide evidence for a critical role for N methyl-D-aspartate (NMDA) receptor activation in LTP induction in this preparation, and demonstrate that this LTP is input-specific. Control pathways showed no potentiation effects. LTP was found in a monosynaptic and two polysynaptic components (average latencies to peak: 8.1, 15.2 and 20.0 ms) and in the superimposed population spikes. Although LTP could be induced with one train per day or with low-intensity trains, larger and longer-lasting potentiation effects could be induced by increasing the number of trains delivered per session, the number of sessions over which trains were delivered, or the pulse intensity of the trains. The LTP decayed slowly and was still evident 5 weeks later. Administration of the competitive NMDA antagonist 3-[(+/-)-2 carboxypiperazin-4-yl]-propyl-1-phosphonic acid blocked the induction of LTP in a dose-dependent fashion and appeared to unmask a depression of both the population spikes and a polysynaptic component. These results indicate that the neocortex is highly sensitive to LTP induction procedures, as long as the stimulation trains are spaced and applied over a period of days. They are also consistent with the view that the neocortex must operate with a slow learning rate to reduce interference effects in memory. PMID- 9863700 TI - Changes in field potentials and membrane currents in rat sensorimotor cortex following repeated tetanization of the corpus callosum in vivo. AB - Repeated, daily tetanization of the corpus callosum induces lasting changes in sensorimotor cortex field potential responses, but the synaptic populations that mediate these responses and support long-term potentiation (LTP) have not been characterized. Current source density analyses of field responses were compared between control animals and those in which LTP was induced by 10 daily series of tetanizations. Tetanization and paired-pulse stimulation (100 ms interval) enhanced the duration of initial (approximately 3 ms onset) deep-negative population spike activity generated by a current sink in layer V that peaked repeatedly at a frequency of approximately 400 Hz. The early (approximately 10 ms to peak) surface-negative component of field responses was generated by a current sink in upper layer V and a source in layer VI. This monosynaptic component followed high stimulation frequencies, recovered quickly from the effects of anaesthesia, and was enhanced by both tetanization and paired-pulse stimulation. The late (approximately 20 ms to peak) surface-negative component was generated by a sink in upper layer V and a source deep in layer V, and was greatly enhanced by tetanization and paired-pulse stimulation. The late component did not follow high-frequency stimulation and recovered slowly from anaesthesia, suggesting that it is driven polysynaptically. Potentiation of monosynaptic thalamic and cortico cortical afferents probably mediates enhancements of the early component and population spikes, while potentiation of polysynaptic afferents to layer V may contribute to growth in the late component. PMID- 9863701 TI - Distribution of cortical activation during visuospatial n-back tasks as revealed by functional magnetic resonance imaging. AB - Human neuroimaging studies conducted during visuospatial working memory tasks have inconsistently detected activation in the prefrontal cortical areas depending presumably on the type of memory and control tasks employed. We used functional magnetic resonance imaging to study brain activation related to the performance of a visuospatial n-back task with different memory loads (0-back, 1 back and 2-back tasks). Comparison of the 2-back versus 0-back tasks revealed consistent, bilateral activation in the medial frontal gyrus (MFG), superior frontal sulcus and adjacent cortical tissue (SFS/SFG) in all subjects and in six out of seven subjects in the intraparietal sulcus (IPS). Activation was also detected in the inferior frontal gyrus, medially in the superior frontal gyrus, precentral gyrus, superior and inferior parietal lobuli, occipital visual association areas, anterior and posterior cingulate areas and in the insula. Comparison between the 1-back versus 0-back tasks revealed activation only in a few brain areas. Activation in the MFG, SFS/SFG and IPS appeared dependent on memory load. The results suggest that the performance of a visuospatial working memory task engages a network of distributed brain areas and that areas in the dorsal visual pathway are engaged in mnemonic processing of visuospatial information. PMID- 9863702 TI - Effects of neonatal freeze lesions on expression of parvalbumin in rat neocortex. AB - Neonatal freeze lesions to the cortical plate result in focal malformations of the cerebral cortex that resemble four-layered microgyria. These malformations have been associated with local and distant changes in neuronal architecture, and have been implicated in the neocortical epileptiform discharges that can spread up to 4 mm away from the malformation itself. In an effort to assess potential changes in the development of one population of inhibitory interneurons in this malformation, we measured the density of parvalbumin-immunoreactive (ParvIR) neurons in microgyric and control cerebral cortex on postnatal days 13, 15, 21 and 64. In comparison to controls, microgyric animals exhibited a transient decrease in the expression of parvalbumin immunoreactivity in supragranular neurons, both within the malformation itself and in normal six-layered cortex up to 2 mm adjacent to it. This difference disappeared by P21. In addition, there was a permanent diminution of the density of ParvIR neurons in infragranular layers both within and immediately adjacent to the microgyrus. These results indicate that early injury to the cortical plate gives rise to both focal and more widespread changes in cortical architecture. PMID- 9863703 TI - Congenital deficiencies and abnormalities of prothrombin. AB - Prothrombin (factor II) deficiency was first described in 1947 by Quick et al., although the first prothrombin abnormality was reported in 1969 by Shapiro et al. The condition is still considered very rare. In spite of its rarity, the defect has allowed important improvements in our understanding of both congenital and acquired prothrombin deficiencies. The diagnosis of prothrombin deficiency or abnormality can be made using a combination of clotting, chromogenic and immunological assays. In cases of true deficiency, a parallel decrease in all these assays is observed, regardless of the activating agent. If discrepancies among the clotting assays are noted, particularly using viper venoms, a dysprothrombinemia should be suspected. Usually, activity levels less than 10% of normal are found in homozygotes, and between 40 and 60% in heterozygotes. Factor II levels in congenital dysprothrombinemias are more variable since one may encounter homozygotes, heterozygotes and compound heterozygotes between a heterozygous abnormality and heterozygous 'true' deficiency or between two distinct abnormalities. Usually the levels of factor II vary between 1 and 50% of normal. Antigen levels in congenital dysprothrombinemias will be normal, near normal or slightly decreased but always higher than the clotting counterpart. Cases with a parallel decrease in prothrombin activity and antigen should not be considered as examples of hypoprothrombinemia. The gene involved in the synthesis of prothrombin is located in chromosome 11. It is composed of 10 exons and 8 introns. Molecular biology studies have discovered several point mutations in some of the dysprothrombinemias. Bleeding manifestations may be severe in homozygous 'true' deficiency and may be more variable in dysprothrombinemias. Heterozygotes are usually asymptomatic. Prognosis is variable and generally in agreement with the prothrombin activity level. In homozygous true deficiency, hemarthroses and intracranial bleeding have been described. Substitution therapy is based on the administration of prothrombin complex concentrates or of plasma. The long half-life of prothrombin injected, about 70 h, allows the achievement of hemostatically effective levels (about 50% of normal) without difficulty. PMID- 9863704 TI - Comparative effects of synthetic pentasaccharide, low-molecular-weight heparin, unfractionated heparin and recombinant hirudin on the generation of factor VIIa and prothrombin activation after coagulation of human plasma. AB - We studied the effect of synthetic pentasaccharide, a low-molecular-weight heparin (enoxaparin), unfractionated heparin and recombinant hirudin on the generation of factor VIIa (FVIIa) and prothrombin activation after in-vitro clotting of human platelet-poor plasma. FVIIa was measured with a new clotting assay that uses recombinant tissue factor truncated to interact only with FVIIa. Residual prothrombin was measured using the conventional clotting assay. FVIIa and residual FII were measured in the liquid - called pseudo-serum (psi-serum) - obtained 1 h after clotting of normal platelet-poor plasma. A kinetic study of the generation of FVIIa was also performed. Coagulation was initiated by triggering the extrinsic, the intrinsic and both associated clotting pathways. Levels of FVIIa in the psi-sera (55+/-15, 258+/-18, and 164+/-18 ng/ml, in the extrinsic, intrinsic and intrinsic + thromboplastin psi-serum respectively; values are means+/-SEM) were significantly increased compared with those in the platelet-poor plasma (3 ng/ml). Pentasaccharide, low-molecular-weight heparin and unfractionated heparin inhibited the generation of factor VIIa or its activity, or both, in a dose-dependent manner in all the experimental systems (60-90% inhibition). A kinetic study revealed that the inhibition of the generation of FVIIa by pentasaccharide and heparins starts 1 min after triggering either the extrinsic or the intrinsic clotting pathway. The downregulation of FVIIa by heparins was effected mainly by their anti-Xa activity, but also by their inhibitory effect on the generation of prothrombinase. Pentasaccharide, enoxaparin and unfractionated heparin significantly inhibited prothrombin activation in both extrinsic and intrinsic experimental system. Hirudin had no inhibitory effect either on the generation of FVIIa or on prothrombin activation in any experimental system. PMID- 9863705 TI - Markers of coagulation and platelet activation during percutaneous mitral valvuloplasty. AB - The effect of percutaneous mitral valvuloplasty (PMV) on markers of coagulation and platelet activation was investigated to assess whether coagulation or platelet activation take place during and after PMV even under conditions of full heparinization. Before PMV, all the hemostatic parameters studied were in the normal range compared with those of a control group. Two hours after PMV the levels of prothrombin fragment F1 and 2 (F1+2) (1.6+/-0.6 nmol/l versus 0.8+/-0.3 nmol/l, P < 0.005), plasma thrombin-antithrombin III (TAT) complexes (5.4+/-3.2 ng/ml versus 2.2+/-0.9 ng/ml) and beta-thromboglobulin (119+/-70 ng/ml versus 42.2+/-41 ng/ml) had increased significantly compared with those measured at basal conditions. Activated partial thromboplastin time was significantly prolonged (152+/-40 s versus 21+/-5 s), reflecting full heparinization. Levels of fibrinogen, F1+2, TAT and beta-thromboglobulin remained increased 72 h after PMV. We conclude that patients with severe, symptomatic mitral stenosis undergoing PMV need a more specific antithrombotic therapy or a more prolonged and perhaps less intensive heparinization. PMID- 9863706 TI - A novel antithrombin-heparin covalent complex: antithrombotic and bleeding studies in rabbits. AB - Heparin has been used extensively for prophylaxis and treatment of deep vein thrombosis. However, heparin has several limitations including a short intravenous half-life, inability to inhibit clot-bound thrombin, and bleeding. We have developed a covalent antithrombin-heparin complex (ATH) that has a longer intravenous half-life and greater anticoagulant activity than heparin. The antithrombotic activity of ATH was tested in a rabbit jugular vein thrombosis treatment model. Administration of ATH caused a 17% reduction in clot weight compared with an increase of 24, 60, 172 and 135% for administration of antithrombin plus heparin, heparin, antithrombin and saline, respectively. Clot weight and fibrin accretion were both significantly lower in the ATH group than in the antithrombin plus heparin group (P < 0.05). The peak anti-factor-Xa activity was fourfold higher in the ATH group than in the antithrombin plus heparin group. Using a rabbit bleeding ear model, there was no significant difference in cumulative blood loss between ATH and antithrombin plus heparin groups, at similar plasma anti-factor-Xa levels. In conclusion, ATH has superior antithrombotic activity and similar bleeding effect compared with heparin on a mass basis. The enhanced antithrombotic activity of ATH may be a result of its increased anticoagulant activity or its longer half life, or both. PMID- 9863707 TI - Plasminogen activator inhibitor-1 4G/5G-polymorphism and factor V Q506 mutation are not associated with myocardial infarction in young men. AB - Several recent studies have reported contradicting results concerning the relevance of the plasminogen activator inhibitor-1 (PAI-1) 4G/5G-polymorphism for myocardial infarction. In addition, the common factor V Q506 (FV:Q506) mutation is frequently discussed as a risk factor for arterial thrombosis, but evidence is rare. In order to further highlight the role of both polymorphisms in myocardial infarction, we investigated 241 young male myocardial infarction patients (< or = 45 years-of-age) aged 38.6+/-4.4 years (mean+/-SD) for the presence of both genotypes. The control group consisted of 179 healthy men aged 47.1+/-6.4 years (mean+/-SD) of the same ethnic background as the patients. Neither the distribution of the PAI-1 4G/5G-polymorphism nor the prevalence of the FV: Q506 mutation was significantly different between young patients and controls (4G/4G genotype: chi2=2.08, NS; odds ratio 1.36, 95% confidence interval 0.89-2.06; FV:Q506 mutation: chi2=0.33, NS; odds ratio 1.33, 95% confidence interval 0.64 2.78). Moreover, the PAI-1 4G/5G-distribution did not differ significantly between patients and controls in subgroups by tertiles of triglyceride levels. In conclusion, in the present study neither homozygosity for the 4G allele of the PAI-1 4G/5G-polymorphism nor the FV:Q506 mutation led to an increased risk of myocardial infarction in young men. PMID- 9863708 TI - Diagnostic value of two rapid and individual D-dimer assays in patients with clinically suspected pulmonary embolism: comparison with microplate enzyme-linked immunosorbent assay. AB - A semiquantitative enzyme-linked immunosorbent assay (ELISA) test (Instant IA D dimer) and a new quantitative rapid and individual test (STA-Liatest DDi) were compared with the reference microplate ELISA (Asserachrom D-Di) for D-dimer testing in 142 patients clinically suspected of pulmonary embolism, on the basis of clinical symptoms and signs, electrocardiogram, blood gases and chest X-Ray abnormalities. The cut-off value for the quantitative tests was 500 ng/ml and Instant IA was interpreted by three readers. Pulmonary embolism was confirmed by lung scan or angiography in 60 patients (42%). The sensitivities of ELISA and STA Liatest DDi were 92% [95% confidence interval (CI) 82-97%] and 93% (95% CI 84 98%), respectively. The three readings of Instant IA D-dimer disagreed in 27 (19%) of the patients and sensitivity varied from 83 to 93% according to the readers. In the 115 patients with concordant readings, sensitivity was 92% (95% CI 82-98%). These results suggest that STA Liatest DDi may be used instead of microplate ELISA for the exclusion of pulmonary embolism, whereas the use of Instant IA D-dimer for this purpose is limited by the number of discordant results. PMID- 9863710 TI - Analysis of genetic polymorphisms related to thrombosis and other risk factors in patients with retinal vein occlusion. AB - The purpose of this study was to investigate the role of genetic polymorphisms associated with venous and arterial thrombosis in patients with retinal vein occlusion (RVO). One-hundred and two consecutive patients with RVO were examined for factor V G1691A and factor II G20210A, methylenetetrahydrofolate reductase (MTHFR) C677T and apolipoprotein E4 by amplification of specific DNA fragments and restriction analysis. The risks exerted by these polymorphisms and by the conventional risk factors of RVO were evaluated by comparing their frequencies among patients and controls and by estimating the respective odds ratios. We found that the prevalences of the factor V G1691A, factor II G20210A, and apolipoprotein E4 polymorphisms were similar in the study and control groups. Logistic regression analysis involving the parameters for which significant differences were detected disclosed an odds ratio of 1.9 for MTHFR C677T homozygosity (95% confidence interval 0.95-3.81), an odds ratio of 2.12 for hypertension (95% confidence interval 1.16-3.73) and an odds ratio of 3.25 for a family history of stroke (95% confidence interval 1.07-9.51). Our data suggests that homozygosity for the MTHFR C677T polymorphism is a risk factor of RVO in addition to arterial hypertension and a family history of stroke. PMID- 9863709 TI - In-vitro effect of oncostatin M on the release by endothelial cells of von Willebrand factor, tissue-type plasminogen activator and plasminogen activator inhibitor-1. AB - Epidemiological studies have demonstrated that levels of plasma fibrinogen, von Willebrand factor (vWf), plasminogen activator inhibitor-1 (PAI-1) and tissue type plasminogen activator (tPA) are associated with the incidence of vascular disease. Since oncostatin M dramatically induces fibrinogen biosynthesis by hepatocytes and could be implicated in vascular injury leading to atherosclerosis, we have analyzed the effect of oncostatin M on PAI-1, vWf and tPA secretion by endothelial cells. A 2-h incubation of human umbilical vein endothelial cells with oncostatin M increases thrombin-induced secretion of vWf to the same extent as tumour necrosis factor-alpha or interleukin-1 (137+/-26% of control for 5 ng/ml oncostatin M, P < 0.001, n=5). The effects on tPA and PAI-1 secretion were different depending on the type of endothelial cells tested. On human umbilical vein endothelial cells, oncostatin M induced an increase in PAI-1 and a decrease in tPA secretion, which could explain the thrombogenicity of oncostatin M on large vessels. On a human microvasculature endothelial cell line, oncostatin M did not modify PAI-1 but induced an increase in tPA secretion. This observation of the effects of oncostatin M on both macro- and microvasculature could explain the increased levels of vWf, PAI-1 and tPA in the plasma of atherosclerotic subjects identified in epidemiological studies, suggesting that oncostatin M could play a key role in the development of atherosclerotic lesions. PMID- 9863711 TI - Acquired haemophilia A: errors in the diagnosis. AB - The distinction between a specific factor inactivator and a non-specific inhibitor is important when confronted by a patient with a history of bleeding and abnormal in-vitro coagulation tests. We report on two patients who presented with bleeding and a prolonged activated partial thromboplastin time. Initial factor assays suggested combined deficiency of factors VIII and IX as a result of the presence of inactivators. The use of dilution studies, chromogenic assays, a novel in-house enzyme-linked-immunosorbent-assay-based technique and phospholipid neutralization, demonstrated that Case 1 had a genuine factor VIII inactivator resulting in factor VIII levels of less than 1 IU/dl but no factor IX deficiency. Case 2 had normal levels of factor VIII on further testing and no specific inactivator to either factor VIII or IX but a potent antiphospholipid antibody which had interfered with the phospholipid-dependent in-vitro assays. Care must be taken in the interpretation of laboratory assays in the presence of antiphospholipid antibodies to ensure that the correct diagnosis is made and inappropriate treatment avoided. PMID- 9863712 TI - Effect of zinc ions on fibrin network structure. AB - In addition to calcium, other physiologically important divalent cations (magnesium and zinc) are known to influence fibrin formation and structure. We have studied the effect of different concentrations (0-20 micromol/l) of zinc ions (Zn2+) in the absence and presence of calcium on the gel structure formed in purified fibrinogen-enzyme systems. For that purpose, we used turbidity measurement, liquid permeation and confocal three-dimensional microscopy of the gel as well as sodium dodecyl sulphate (SDS)-gel electrophoresis. The results of turbidity measurements indicated that the clotting time decreased with increasing concentrations of Zn2+. The fiber mass: length ratio (mu) values showed that the porosity of the gels increased in a concentration-dependent manner, i.e. at higher concentrations of Zn2+, larger pores with thicker fibrin fibers were formed. Three-dimensional microscopy data of the gels were in good agreement with the mu data. On SDS-gel electrophores of reduced fibrin, no cross-linking was observed in the presence of zinc ions only (without the addition of calcium ions), nor were D-D dimer bands observed in non-reduced plasmin digested fibrin samples in the presence of zinc ions only. The above results show that zinc changes the fibrin gel structure and that this effect appears to be independent of calcium. PMID- 9863713 TI - Development of a method to separate lipoprotein-bound and lipoprotein-depleted tissue factor pathway inhibitor. Measurement of free tissue factor pathway inhibitor activity. AB - The aim of this study was to develop a method to separate lipoprotein-bound from lipoprotein-free tissue factor pathway inhibitor (TFPI) and to measure the TFPI chromogenic activity and antigen in both fractions. This was performed by ultracentrifugation of plasma, after increasing its density to 1.21 g/ml with potassium bromide. Blood was taken from nine volunteers before and after an injection of low-molecular-weight heparin. The ultracentrifugation procedure was adequate, since the mean cholesterol recovery was 91% and only 2% of the cholesterol remained in the lipoprotein-depleted fraction. No free TFPI protein was found in the lipoprotein-rich fraction. Moreover, the amount of free TFPI in the lipoprotein-depleted fraction was close to that found in plasma. Using this method, we confirmed that heparin does not induce an increase in bound TFPI and that the moderate increase in total TFPI antigen in plasma is entirely caused by the enhancement of free TFPI. We then applied the TFPI chromogenic assay to the lipoprotein-depleted fraction to assess the activity of free TFPI. The activity was 0.11+/-0.03 and 0.36+/-0.08 U/ml before and after heparin, respectively (a 3.6-fold increase) while the activity of bound TFPI did not increase at all. We suggest that this method may be an alternative to gel filtration for measuring free TFPI activity, and might be of value in the search for TFPI abnormalities in patients with thrombosis. PMID- 9863714 TI - Effect of plasma pooling on the International Sensitivity Index of prothrombin time systems. AB - Guidelines set by the World Health Organization (WHO) state that in order to calibrate a prothrombin time system for International Sensitivity Index (ISI), freshly prepared specimens from 20 normal individuals and 60 patients receiving coumarin are required. These numbers are required to obtain a precise value of the calibration line slope when there is considerable scatter of individual data about the regression line. The scatter can be reduced by pooling individual plasma samples. In the present study, four pooled plasmas were prepared, one from 20 normal individuals and three from three groups of 30 patients receiving treatment with long-term oral anticoagulants. Prothrombin times were determined with four thromboplastins, HepatoQuick (rabbit thromboplastin combined with adsorbed plasma), Recombiplastin (recombinant human thromboplastin), Thromborel-S (human placenta), and Thromboplastin-C Plus (rabbit brain). Calibration line slopes were calculated for the six possible combinations of thromboplastins using the set of all individual plasma samples and the set of four pooled plasmas. In most comparisons, the WHO calibration model was appropriate, i.e. the line calculated for the patients' samples passed through the mean of the normals. The calibration line slopes obtained with the set of four pooled plasmas did not differ by more than 5% from the corresponding slopes calculated with the original individual plasmas. For some combinations of thromboplastins non-linear relations were observed both with the individual plasmas and with the pooled plasmas. We conclude that pooling individual plasmas does not significantly change the calibration relation between prothrombin times determined with the original individual plasmas. Freshly pooled plasmas can be used to determine the ISI of prothrombin time systems with an acceptable degree of precision. PMID- 9863715 TI - Platelet activation in patients with antiphospholipid syndrome. AB - The binding of antiphospholipid antibodies to circulating platelets and the potential association with thrombocytopenia and platelet activation was investigated in 25 patients with primary antiphospholipid syndrome (APS). Fourteen patients had a platelet count above 150 x 10(9)/l, and 11 patients had mild to moderate thrombocytopenia of 50-150 x 10(9)/l. The presence of platelet autoantibodies was investigated by immunofluorescent binding. No correlation between the presence of autoantibodies on platelets and thrombocytopenia was found. The binding of antibodies in patients' serum and platelet eluates was investigated by performing enzyme-linked immunosorbent assays with phospholipids as antigens. In seven patients antibodies to negatively charged phospholipids were present in platelet eluates. Platelet activation was measured by flow cytometry using a fluorescein isothiocyanate (FITC) labeled monoclonal antibody to P-selectin (CD62). The binding of anti-P-selectin to patients' platelet surface P-selectin was not increased, compared with the binding to platelets obtained from normal donors. Platelet serotonin concentration in APS patients was significantly lower than that found in the platelets of normal controls. More studies are necessary to determine the exact role of antiphospholipid antibodies in the pathogenesis of thrombocytopenia, and to elucidate the cause of low serotonin levels in platelets of APS patients. PMID- 9863716 TI - Emphasis on congenital conditions predisposing to thrombosis should not make us disregard the importance of acquired factors. PMID- 9863717 TI - Factor V variants, activated protein C resistance and venous thromboembolism. PMID- 9863719 TI - The anticoagulant effect of antithrombin/heparin complex. A new screening test to detect a thrombophilic tendency? PMID- 9863718 TI - The 20210 A allele of the prothrombin gene is not a risk factor for juvenile stroke in the Danish population. PMID- 9863720 TI - Recurrent arterial thrombotic disease on young onset and protein S deficiency. PMID- 9863721 TI - Abstracts of the meeting of the XVth International Fibrinogen Workshop. Cleveland, Ohio, USA. August 13-15, 1998. PMID- 9863722 TI - Review of AAPM Task Group No. 43 recommendations on interstitial brachytherapy sources dosimetry. American Association of Physicists in Medicine. AB - In 1995, the American Association of Physicists in Medicine (AAPM) Task Group No. 43 (TG-43) published its recommendations on the dosimetry of interstitial brachytherapy sources. The report recommended the use of a new dose calculation formalism based on measured quantities. The formalism in modular form permits the computation of doses in two dimensions for 103Pd, 125I, and 192Ir sources. The TG 43 dose calculation formalism introduced new and updated quantities such as air kerma strength, dose rate constant, radial dose function, anisotropy function and anisotropy factor. The dose rate obtained using the TG-43 dose calculation formalism and updated source dosimetry data can be expected to be different from some of the currently used systems by as much as 17%. For the same treatment and implementing the TG-43 dosimetry with point source approximation, the widely prescribed dose of 160 Gy for 125I permanent implants using model 6711 sources changes to 144 Gy. In addition to the dose calculation formalism, TG-43 report also stated that the air kerma strength provided by NIST is estimated to be approximately 7-10% higher than it should be, due to low energy photon contamination for 125I. This difference has not been accounted for in the TG-43 report. PMID- 9863723 TI - Using static MLC fields to replace partial transmission cerrobend blocks in treatment planning of rectal carcinoma cases. AB - A treatment planning technique has been developed for using static multileaf collimators to replace partial transmission blocks for treating rectal or cervix carcinoma. The static MLC fields were used to replace the partial transmission block of the anterior-posterior pelvis field for the so-called "thunderbird" technique. Treatment plans were developed and evaluated on a commercial three dimensional treatment planning system (FOCUS, Computerized, Medical Systems, St. Louis, MO). The result of the treatment plan comparison indicates that the static MLC fields are capable of achieving the same target, inguinal and pelvic dose distribution as the partial transmission cerrobend blocked fields. The MLC fields are easy to modify particularly for the match line adjustments. In conclusion, it is efficient and effective to use static MLC fields to replace partial transmission blocks in the "thunderbird" technique for treating rectal or cervix carcinoma. PMID- 9863724 TI - Clinical considerations in the use of missing tissue compensators for head and neck cases. AB - The irregular shape or contour of the patient's surface in the treatment field can alter the dose distribution resulting in non-uniformity of dose in the treatment volume. Missing tissue compensators have been most commonly used to improve this non-uniformity, especially in head & neck, breast, lung and supraclavicular regions. Two or three dimensional compensators have been typically designed to make the dose uniform at a specific depth. This compensation shifts the dose distribution within the treatment volume so that some structures may be under or over compensated. This study will examine how various sites in head and neck cases are affected by compensators. We have also analyzed the uncertainty in compensated dose due to the daily variations in patient repositioning. Computer isodose plans using Cobalt-60 gamma rays and 6 and 18 MV x-rays were generated using coronal contours. Results show that the dose uniformity is improved for the treatment sites, especially for the thinner sites, like the larynx and the anterior cervical neck nodes. Finally, patient movement or positioning errors of +/-1.0 cm will cause a change in dose distribution. PMID- 9863725 TI - A review of digital image networking technologies for radiation oncology treatment planning. AB - Dosimetrists, physicists, and physicians rely heavily upon digital imaging modalities in their daily work. In the current healthcare marketplace, many radiotherapy facilities operate satellite centers or may be undergoing mergers with other service providers in their community. As a consequence, the development of network-based radiotherapy image communication may yield a significant impact on the clinical operation of such centers. Digital image networking will become an everyday tool in radiotherapy treatment planning in the near future. As responsible users of this technology, an accurate perception of what the network does is essential and enhances our ability to utilize it. This article presents a review of the network architecture and transmission standards necessary for understanding and developing a radiotherapy image network. PMID- 9863726 TI - User introduced uncertainties in the measurement of field misalignment by a computer aided technique. AB - While images acquired using an electronic portal imager may be visually compared to other portal or simulator images to assess patient alignment, a more quantitative comparison is desirable. Computer aided alignment tools are available which are based on user placed landmarks. The purpose of this study was to estimate uncertainties introduced by the user to the final measurement of field misalignment using such a tool. Both intra- and inter-user reproducibility were assessed. To complete this task, a number of image pairs including both phantom and patient images were compared by multiple observers. Results of the comparisons (x- and y- translation and rotation) were tabulated for each image pair and their reproducibility assessed by calculating a mean and standard deviation. User introduced uncertainty was found to be independent of the magnitude of rotation or x- or y- translation. In all cases, there was no difference between intra-observer and inter-observer uncertainty. For clinical cases, there is a significant difference between uncertainty in x- and y translation due to both image quality and patient anatomy. In addition, the magnitude of uncertainty tracks qualitatively with image quality and number of available anatomical landmarks. The decision to make a correction in field alignment must be made considering these uncertainty estimates. Image comparison must be fully automated to eliminate uncertainties introduced by the user. PMID- 9863727 TI - Dosimetric comparison of centered and off-centered posterior neck electron fields. AB - Approximately rectangular low or medium energy electron fields at extended SSD are often used to boost over the spinal cord in the treatment of head and neck cancer after cord tolerance is reached. A separate abutting photon field is used to continue treatment anterior to the spine. Typically, the electron and photon fields have different central axes and the electron cutout is symmetrically centered in the cone. However, a good match between the photon and electron fields is achieved more readily if the central axis of the electron field is located at the center of the area treated by the photon field. This displaces the electron cutout toward the edge of the cone. We measured and compared the percent depth dose (PDD), output factors and profiles for matched pairs of centered and off-centered rectangular and square cutouts for the 6, 9 and 12 MeV beams of three Varian linacs. The 10 x 10 and 15 x 15 cones were used at SSDs of 100 cm and 110 cm. Differences between centered and off-centered cutouts of the same dimensions were less than 1% for PDD and profiles and less than 5% for output factors. Therefore, the same central axis can be used for abutting photon and electron fields, without requiring extra dosimetric data to account for the off center location of the electron cutout. PMID- 9863728 TI - Proton beam output measurement with an extrapolation chamber. AB - A variable air-volume, parallel-plate extrapolation chamber forming an integral part of a polystyrene phantom was used in measurement of dose rate in a 250 MeV clinical proton beam. The sensitive air-volume of the extrapolation chamber is controlled through the movement of the chamber piston by means of a micrometer mounted on the phantom body. The relative displacement of the piston is monitored by a calibrated mechanical distance travel indicator. The proton beam dose rate determined with the uncalibrated extrapolation chamber was 5% lower than the dose rate determined with a calibrated Farmer-type thimble chamber at the same depth in the polystyrene phantom. Despite the current 5% discrepancy, uncalibrated extrapolation chambers may offer a simple and practical alternative to current techniques used in output measurements of proton beam machines. PMID- 9863729 TI - Dosimetric considerations of water-based bolus for irradiation of extremities. AB - The dosimetry of high-energy photon beams in the treatment of superficial lesions occurring in extremities was examined. Large parallel-opposed fields with different photon beam energies were used. The extremity was immersed in water contained in a commercially available plastic wastebasket. The water bolus serves to even out the surface irregularities of the extremities and to remove the skin sparing effect. A polystyrene block was placed at the floor of the wastebasket to ensure that the extremity was encompassed in the radiation fields. The photon beam energies considered were 4 MV, 6 MV, 10 MV, and 24 MV. The results show that the dose distributions are more homogeneous with higher photon beam energies. The isodose lines are more constricted at mid-plane for low energy photon beams. Higher energy photon beams, 10 MV and up would be preferable for the treatment of superficial lesions of the extremities immersed in water bolus contained in a typical wastebasket size. PMID- 9863730 TI - Effects of collimator jaw setting on dose output for treatments with multileaf collimator. AB - The variations in dose output with collimator jaw setting have been investigated for treatment fields shaped by a multileaf collimator. Measurements have been made for 6 and 15 MV x-ray beams on a Varian Clinac 2100C machine. The results of our study show that the collimator jaw settings can affect the dose by about 5% for a small field shaped by a multileaf collimator. The effect is smaller for larger fields. PMID- 9863731 TI - Partial lung shield for TBI. AB - In this study, AP/PA setup was used with a 4 MV linear accelerator. The prescribed dose is 1000 cGy, with a dose per fraction of 200 cGy. One of the simplest lung dose determinations is the nomograph relating dose correction factor and patient thickness. To save time in taking port film in every treatment, lung block is used in alternating fashion: anterior blocks are used in the second and fourth fraction and posterior block is used in the third and fifth fraction. If we set the average correction factor for the lower density lung to 1.18, the six open fields will deliver an average of 708 cGy to the lung. To limit the lung dose to 10 Gy, each blocked field should deliver 73 cGy. Measurements showed that a thickness of slightly less than 1 cm of cerrobend provides approximately 65% transmission, which approximates the desired 73% broad beam transmission. The proper transmission of the partial lung shield is verified by measurements made for the patients with diodes placed at the entrance and the exit side of the lung. While we describe a particular setup, the procedure can be modified to accommodate different dose fraction. There are many ways to deliver the TBI dose, and the method of constructing the partial lung shield described here is one of the many methods that may be used to limit the dose to the lung. PMID- 9863732 TI - Clinical commissioning of Laitinen Stereoadapter for fractionated stereotactic radiotherapy. AB - The Laitinen Stereoadapter 5000 from Sandstroem Trade and Technology was acceptance tested and commissioned for clinical use in a Fractionated Stereotactic Radiotherapy Program at our facility. The frame was implemented to function as a localization device for target delineation rather than as an immobilization device. The frame is of non-invasive nature utilizing ear plugs and a nasion bridge adapter as the connecting points with the patient's head. The reproducibility of the head frame position with respect to external skull reference points was tested. CT and MRI imaging studies were performed on a patient phantom with the stereoadapter in place. The target was delineated and target coordinates were calculated for two implanted targets. The phantom was positioned according to the target coordinates on a Siemens MXE Linear Accelerator by aid of the target positioning lasers. Radiographic port film images were taken with the circular fields typically used in stereotactic radiosurgery. A complete treatment isodose plan was performed and dosimetric accuracy was tested by positioning a small volume ionization chamber at the center of the target volume in the head phantom. The results of these tests were found to be clinically acceptable. PMID- 9863733 TI - Standard linear plans in single channel high dose rate brachytherapy: a dosimetric analysis. AB - The use of standard linear plans is proposed for single channel intraluminal High Dose Rate brachytherapy. This technique employs the optimized dwell times derived from a straight line within a curved geometry. Such standardization of the planning procedure ensures expedient delivery of treatment. The 3-D dose distribution resulting from the use of standard linear plans within various curved geometries is investigated. In this study a comparison is made between the dose delivered to the perimeter of the target volume from both standard linear plans and individually optimized plans. Our results demonstrate that the use of a standard linear plan is acceptable in curved geometries, given the current practice of dose and volume specification for high dose rate intraluminal brachytherapy. PMID- 9863734 TI - Medical dosimetry training curriculum. American Association of Medical Dosimetrists. PMID- 9863735 TI - A code of ethics for the medical dosimetrist--the American Association of Medical Dosimetrists experience. AB - The Committee on Protocol and Ethics of the American Association of Medical Dosimetrists (AAMD) has developed a Code of Ethics for a radiation oncology society of medical dosimetrists. The purpose of the code of ethics is fourfold: (1) Establish an ideal of professional conduct specific to the medical dosimetry profession; (2) Develop a statement of the moral values and commitment of the AAMD; (3) Recognize professional relationships and obligations; and (4) Define goals to which the medical dosimetrist should aspire. The Code of Ethics was adopted as AAMD policy in October 1995. PMID- 9863736 TI - Reversal of "no reflow" during vein graft stenting using high velocity boluses of intracoronary adenosine. AB - Slow or no reflow is a serious problem complicating catheter-based revascularization of degenerated saphenous vein bypass grafts. We examined the efficacy of rapidly delivered, high-velocity injections of intracoronary adenosine to reverse 11 slow-flow events complicating stenting of diseased bypass grafts. Ten of 11 events were rapidly improved to TIMI 3 flow by this technique within 3.8+/-1.6 min of the initial adenosine injection. In an ex vivo model, 3 ml syringes created higher peak pressures and velocities than 10- and 20-ml syringes. We conclude that rapid and repeated high-velocity intragraft administration of adenosine is a promising new approach to promptly reverse no reflow events complicating PTCA and stenting of diseased saphenous vein grafts. Ex vivo studies demonstrate a potentially important mechanical advantage with the use of small syringes for injection. Further randomized studies will be required to better define the mechanism(s) and efficacy of this approach for treating no reflow, including its use in native vessels. PMID- 9863737 TI - Adenosine boluses: a cure for "no-reflow"? PMID- 9863738 TI - Initial and follow-up results of the ACS multi-link stent: a single center experience. AB - The Palmaz-Schatz (PS) stent has effectively reduced restenosis; however its rigidity makes it sometimes difficult or impossible to deliver. The initial and follow-up outcomes with the ACS Multi-Link stent (deployed from April to November 1995) were evaluated in 70 patients (79 lesions): unplanned in 34% (abrupt closure 1%; threatened closure 5%; suboptimal results 28%) and planned in 66%. Three to six month follow-up angiograms were analyzable in 67 lesions; 96% procedural (in nine lesions PS stenting had failed) and 95% clinical success were achieved. In-hospital mortality was 1.4%. Myocardial infarction occurred in 2.9%, and subacute stent thrombosis in 1.4%. Stenting improved immediately the minimal luminal diameter (from 0.97+/-0.41 mm to 2.72+/-0.31 mm), but at 6 months it had decreased to 1.89+/-0.44 mm. Angiographic restenosis (<50% diameter stenosis) occurred in 11, a rate of 16.4%; target lesion revascularization (TLR) was required in six (re-PTCA in five or bypass surgery in one; 6/67=8.7%). Actuarial 1-2 year survival rate was 91%, 80% surviving free from major complications or need for TLR. We conclude that the ACS Multi-Link stent can be implanted in lesions unsuited for the PS stent with a high success rate, and an anticipated restenosis rate perhaps comparable to with the PS stent. PMID- 9863739 TI - Coronary stents: increasing options and indications. PMID- 9863740 TI - Safety of coronary arteriography in clinically stable patients following coronary bypass surgery. Post CABG Clinical Trial Investigators. AB - The frequent use of diagnostic coronary arteriography and its importance in evaluating results of intervention in clinical trials emphasize the necessity of continued assessment of procedural risk. Several studies have described such risks, but they have often included a diverse group of patients with varying levels of clinical stability. Furthermore, this risk has not been well established in a population of patients with saphenous vein bypass grafts. There is need to define the risk of coronary arteriography in a group of patients who are both clinically similar and stable, and to evaluate the influence of improved technology and increased operator experience on the risk of the procedure. The National Heart, Lung, and Blood Institute-funded Post Coronary Artery Bypass Graft Trial offered the opportunity to evaluate the risk of elective diagnostic coronary arteriography in clinically stable patients studied at two points in time: pre-enrollment and 4-5 years after study entry. In this group of 2,635 angiograms from clinically stable patients over 5 years there were no deaths and the risk of myocardial infarction was 0.08%, while 0.7% had clinically important complications. Non-elective, urgent studies (311 angiograms) on unstable patients were more likely to include angioplasty and were associated with a risk of death of 0.6% and myocardial infarction of 1.3%. Complications did not vary with age or gender. Vascular trauma was more likely to occur using the brachial than the femoral artery entry sites. These results indicate that elective angiography on stable patients can be accomplished with a very low risk of mortality (0% in this study) or serious cardiovascular complication. This supports the safety and usefulness of angiography for clinical intervention trials. PMID- 9863741 TI - Validation of Doppler FloWire for measurement of coronary flow reserve in humans. AB - Previous studies have validated the 133Xenon (133Xe) method to assess regional myocardial blood flow and coronary flow reserve (CFR). Doppler FloWire (DFW) has been used recently for measuring CFR to assess the physiological significance of coronary stenosis. Data obtained by DFW has never been correlated to 133Xe. Our study compared data from DFW measurement of CFR to that obtained by 133Xe in 31 consecutive patients with variable coronary stenosis. Regional myocardial blood flow was measured by assessing the rate constants of 133Xe washout using multicrystal gamma camera after injection (20 millicuries) in the right or left coronary artery. CFR was assessed by measuring resting and hyperemic coronary blood flow by 133Xe and DFW using i.v. adenosine (140 mcg/k/min x 3 min). CFR was also measured by DFW giving intracoronary (i.c.) adenosine (12 microg in the right coronary, 18 microg in the left). In both methods--133Xe and DFW--coronary flow reserve was defined as the ratio of maximal hyperemic-to-baseline flow. DFW and 133Xe assessment of CFR correlated highly, whether adenosine was used i.c.(r=0.87; P=0.0001) or i.v.(r=0.78; P=0.0001). CFR obtained by DFW following i.c. and i.v. adenosine correlated well (r=0.79; P=0.0001). i.c. adenosine has fewer side effects. Both DFW and 133Xe are comparable in measuring CFR in humans. PMID- 9863742 TI - Rheolytic thrombectomy in the treatment of acute limb-threatening ischemia: immediate results and six-month follow-up of the multicenter AngioJet registry. Possis Peripheral AngioJet Study AngioJet Investigators. AB - We tested the efficacy of rheolytic thrombectomy in treating 21 patients (mean age 68+/-12 years; 66% male) and 22 vessels (limbs) who presented to the hospital within 2 weeks of the development of limb-threatening ischemia. Fifty-two percent had contraindications to use thrombolytics, and 57% had severe comorbidities. All of the vessels were occluded with thrombus on the initial angiogram. Procedural success was achieved in 20 limbs (91%). Three patients expired in the hospital, and one expired at follow-up due to nonvascular causes. Acute limb salvage was achieved in 18 of 19 limbs (95%) in the 18 survivors, and 6-month limb salvage was achieved in 16 of 18 limbs (89%) in the 17 survivors. Rheolytic thrombectomy is effective in restoring immediate blood flow in acute limb-threatening ischemia, especially in high-risk surgical patients or patients with contraindications to thrombolytic therapy. PMID- 9863743 TI - AngioJet thrombectomy catheter for the thrombus-laden lesions. PMID- 9863744 TI - Comparison of enteric-coated aspirin and uncoated aspirin effect on bleeding time. AB - Aspirin therapy is an essential part of the drug regimen for patients with acute myocardial infarction (MI), unstable angina, or after coronary angioplasty and coronary stenting. Recognizing this importance, this study sought to compare the bleeding time in two groups of 10 normal volunteers 4 hr after ingestion of either an enteric-coated aspirin or an uncoated aspirin, assuming that a difference between the two groups could be clinically significant. Defining < or = 8 min as normal, 80% of the uncoated group developed abnormal bleeding times, compared to 10% of the enteric-coated group (P < 0.01). The study demonstrates a significant difference between the two types of aspirin preparations on bleeding times in normal individuals. This strongly suggests that some enteric-coated aspirin preparations may not be as effective as uncoated aspirin in acutely decreasing platelet aggregation. Therefore, uncoated aspirin is recommended in the setting of acute MI, unstable angina, or after percutaneous transluminal coronary angioplasty. PMID- 9863746 TI - Transcatheter recanalization and subsequent stent implantation for the treatment of early postoperative thrombosis of modified Blalock-Taussig shunts in two children. AB - Two babies with duct-dependent cyanotic congenital heart disease were palliated with modified Blalock-Taussig shunts. One patient was diagnosed to have tetralogy of Fallot, and the other patient, tricuspid atresia type Ia. Early postoperative arterial oxygen saturations dropped significantly due to shunt thrombosis. Both patients were successfully treated with angioplasty and stent implantation. PMID- 9863745 TI - Percutaneous transluminal angioplasty of radial artery grafts. AB - The radial artery is being used with increasing frequency to replace the saphenous vein as a coronary artery bypass graft, in the belief that it will provide improved long-term patency. Several centers have confirmed that the early results of surgery using the radial artery seem to be better than those obtained with saphenous grafts. Despite these apparent gains, early failure of the radial artery graft can occur and is frequently associated with symptomatic myocardial ischemia. Percutaneous angioplasty is an alternative to reoperation to treat lesions occurring on radial artery grafts. We report on 4 patients who underwent angioplasty of radial artery grafts. PMID- 9863747 TI - Relief of factitious coarctation following occlusion of large patent ductus arteriosus with Gianturco-Grifka vascular occluder. AB - We report an infant with a large patent ductus arteriosus (PDA) and a hypoplastic aortic isthmus presenting with factitious coarctation due to high ductal flow. Following transcatheter occlusion of the PDA with a Gianturco-Grifka vascular occluder, the aortic gradient resolved, thereby eliminating the need for surgery on the isthmus. PMID- 9863748 TI - Acquired coronary artery-pulmonary artery connection. AB - We report a patient who underwent resection of a mediastinal tumor with postoperative irradiation. Cardiac catheterization fifteen years later demonstrated coronary artery-pulmonary artery fistulas from both the right and left coronary arteries. This case report raises the issue of whether external beam irradiation may have been integral in neovascularization and the development of this acquired abnormality. PMID- 9863749 TI - The coronary connection revisited: a fistula or collateral anastomosis? PMID- 9863751 TI - The cure for a rainy day. PMID- 9863750 TI - Coronary vessel perforation during balloon angioplasty: a case report. AB - Coronary perforation can be managed with prolonged balloon inflations, covered stents, or embolization of the vessel. We report on a case of a balloon-induced perforation of the distal left anterior descending artery, that was sealed by injecting preclotted autologous blood through the balloon catheter lumen at the site of the perforation. The patency of the distal vessel was maintained. PMID- 9863752 TI - Percutaneous balloon aortic valvuloplasty during pregnancy: use of the Inoue balloon and the physiologic antegrade approach. AB - We describe the use of the Inoue balloon to dilate the aortic valve by the physiologic antegrade route during pregnancy. A 27-year-old pregnant woman with severe aortic stenosis presented with progressive dyspnea and presyncope at 26 weeks of pregnancy. She subsequently underwent percutaneous valvuloplasty by the antegrade route utilizing the transseptal puncture. We conclude that percutaneous antegrade balloon valvuloplasty by the Inoue balloon (venous approach) is a safe and effective procedure. It markedly reduces fluoroscopy and is a palliative procedure that allows pregnancy to continue. PMID- 9863753 TI - Aortic valvuloplasty: baby or bathwater revisited. PMID- 9863754 TI - Acute coronary occlusion and in-stent thrombosis in a patient with essential thrombocythemia. AB - We describe a case of essential thrombocythemia in a 34-year-old male who presented with acute anterior wall myocardial infarction and a platelet count of 2,100,000/mm3. Primary percutaneous coronary angioplasty and stenting were performed. Postangioplasty course was complicated by stent thrombosis requiring repeat coronary angioplasty and persistent femoral arterial bleeding that was treated with surgical exploration and repair. The patient was subsequently treated with platelet pheresis, acetylsalicylic acid, ticlopidine, hydroxyurea, and anagrelide without further complications. PMID- 9863756 TI - Coronary stent for variant angina: atypical presentation. AB - Pharmacological therapy remains the treatment of choice for Prinzmetal angina. We report an unconventional approach of coronary artery stenting to treat coronary artery spasm in variant angina refractory to triple drug therapy. Favorable clinical and angiographic results and a negative Ergonovine test, under less aggressive medical therapy, are valuable arguments for stenting. Four-month angiographic follow-up showed absence of intrastent restenosis with a negative Ergonovine test. However, long-term follow-up is necessary before advocating this as a systematic approach. PMID- 9863755 TI - Use of coronary artery stents in the treatment of internal carotid artery stenosis at the base of the skull. AB - The management of high-grade stenosis involving the petrosal segment of the internal carotid artery has been limited to angioplasty alone in the past. However, if angioplasty fails or has less than optimum results, then endovascular stent placement should be considered. We present the first two known cases in which the Multi-Link coronary stent was used for the treatment of internal carotid disease in the petrosal segment. The first patient had symptomatic stenosis that failed angioplasty after seven months and the second patient had a symptomatic lesion that dissected upon angioplasty. The flexibility and low profile characteristics of the stent allowed it to be safely deployed with balloon expansion. No complications occurred and the patients tolerated the procedure. Patients were carefully monitored and no new neurologic events in six months of follow up occurred. These cases reveal the good clinical results of stent placement for surgically-inaccessible lesions of the internal carotid artery in the skull base. PMID- 9863757 TI - Intraarterial ethanol for the management of iatrogenic renal artery pseudoaneurysm. AB - Intraarterial injection of absolute ethanol was employed for the management of a pseudoaneurysm following percutaneous nephrolithotomy. The ethanol was delivered selectively using a coronary angioplasty catheter. This seems to be a potentially useful method for the management of severe hematuria following percutaneous renal procedures. PMID- 9863758 TI - Stenting of a renal artery bifurcation stenosis. AB - Renal artery stenosis involving the renal artery ostium often responds suboptimally to balloon angioplasty. In this context, immediate and long-term results can be optimized by stenting. Occasionally, lesions may involve the midartery segment at a branch point. We report a case of a bifurcation lesion which responded poorly to balloon angioplasty despite the kissing balloon technique. Excellent angiographic result was finally achieved by deploying two flexible coil stents in a "kissing" fashion. The patient's hypertension came under control overnight. At 6-month follow up, he remained normotensive on no medication. Repeat renal angiogram also showed no restenosis. PMID- 9863759 TI - Imaging factors and image optimization. AB - Optimization of dose and image quality is task-dependent. The three main image quality parameters, sharpness, contrast, and noise, are interdependent in radiographic systems. The need to minimize patient dose imposes further couplings between these factors. This is the reason why there are no universally applicable imaging systems. PMID- 9863760 TI - Subselective opacification of a left internal mammary artery graft in a tortuous subclavian artery: use of angioplasty technique and a Tracker catheter. PMID- 9863761 TI - Protection of side-branches in coronary lesions with a new stent design. AB - Side-branches often complicate stenting of coronary lesions. We investigated a new stent, characterized by four wider cells in its center, which can be expanded up to 3.5 mm and which are meant to be placed over the ostium of a major side branch. Forty-seven consecutive patients with lesions involving 48 side-branches received one side-branch stent each. Stent deployment was successful in all patients. Twenty-five side-branches needed additional treatment. Nineteen side branches received a PTCA, and 6 additional side-branches were stented. Postinterventional CK-(creatine kinase) elevation was observed in 3 patients (6%). One additional patient was sent for CABG on the day of the procedure due to loss of a stent intended to be placed into the side-branch. The investigated stent proved to be a safe and effective tool to treat this complex subgroup of stenoses in the presence of favorably preserved flow in the side-branches, with a low incidence of periprocedural complications. PMID- 9863762 TI - Bifurcation stenting: the task ahead. PMID- 9863763 TI - Early experience with the Bard XT stent. AB - The Bard XT stent is a new generation balloon expandable intracoronary stent. It has several unique design advantages. Between October 1996 and November 1997, 127 Bard XT stents of various length were deployed in 93 patients with 109 lesions. According to the American College of Cardiology (ACC) and American Heart Association (AHA) classifications 7 lesions were type A, 38 were type B1, 43 were type B2 and 21 were type C [Ellis et al.: Circulation 82:1193-1202, 1990]. Stent delivery was successful in 98% of attempts. Angiographic success was achieved in 98% of 109 lesions. Procedural success was achieved in 94% of 93 patients. Minimal luminal diameter (MLD) increased from 0.91+/-0.34 mm to 3.03+/-0.44 mm and percentage diameter stenosis reduced from 69.1+/-11.07 to 9.96+/-6.81. Complications occurred in four patients. One patient had intracranial hemorrhage, one patient had subacute thrombosis and two patients died postprocedure. Patients were followed for a period of 1 to 14 months (average 7+/-4 months) for major cardiac events and clinical restenosis. The Bard XT stent is a user-friendly device which provided excellent angiographic results and short-term clinical outcome in selected cases. Further study is required to evaluate effects on restenosis. PMID- 9863764 TI - Balloon mitral valvuloplasty: double vs single transseptal puncture. PMID- 9863765 TI - Heat stress, even extreme, does not induce penetration of pyridostigmine into the brain of guinea pigs. AB - Stress due to forced swimming was recently shown to allow penetration of pyridostigmine (PYR) into the brain of mice. Accordingly, it was suggested that in troops exposed to emotional stress under conditions of war, as during the Gulf War, the BBB may have unexpectedly become permeable to PYR thus leading to an increased frequency of CNS symptoms. In this study, the entry of PYR into the brain was investigated in guinea pigs subjected to different heat stress levels. In a first group, guinea pigs were maintained at room temperature for 2 hours, their core temperature remaining stable at about 39.8 degrees C. In a second group, animals were placed in a climatic chamber in order to keep their core temperature at 41.5 degrees C for 2 hours. In a third group, animals were subjected to a high ambient temperature (42.6 degrees C) during about 2 hours and developed heatstroke symptoms, their core temperature progressively increasing and reaching around 44.3 degrees C. In each group, the stress of the animals was assessed by measuring the increase of plasma cortisol level. PYR (0.2 mg/kg, s.c.) was injected 90 minutes after beginning the experiment. Penetration of the drug into the brain was examined by measurement of acetylcholinesterase (AChE) activity in the cortex, the striatum and the hippocampus of the animals 30 minutes after PYR administration. A passage of this drug into the brain was also evaluated autoradiographically after i.v. injection of tritiated PYR 90 minutes after the beginning of the experiment (100 microCi/animal). Whatever the group examined, no entry of PYR into the CNS could be detected. Exposure to an ambient temperature at 42.6 degrees C for 2 hours resulted by itself in a partial inhibition of cerebral AChE activity. Our results, which agree with previous data obtained in humans exposed to heat stress, are opposite to the recent research showing a central passage of PYR in mice following a forced swim stress test. This demonstrated that the penetration of PYR into the brain of rodents under stress depends on the experimental conditions used (animal species, nature of the stressor, etc.). Extrapolations to humans of results primarily obtained in rodents about central passage of a drug under stress must thus be done very carefully. PMID- 9863766 TI - Mercuric chloride induces a stress response in cultured astrocytes characterized by mitochondrial uptake of iron. AB - Mercury exerts a variety of toxic effects on both neurons and glia. Mercury induces aberrations in microtubules, ion channels and mitochondria presumably by binding to sulfhydryl groups. Indirect evidence further suggests that mercury targeted to mitochondria may induce iron-catalyzed oxygen radical production. We have previously shown that the mitochondria of astrocytes subjected to oxidative stress accumulate redox active transition metals that may catalyze the formation of cytotoxic oxygen free radicals. In the present study we have investigated the effect of mercuric chloride on astrocytes in monolayer culture in order to determine whether mercury accumulates in astrocytic mitochondria and whether mercury exposure triggers a stress response-associated uptake of iron. Our results indicate that mercuric chloride exposure initiates the constellation of changes in mitochondrial structure that typifies the response of these cells to oxidative stress. Energy dispersive Xray microspectroscopy demonstrates that these altered mitochondria concentrate both mercury and iron. Concurrent with these changes, mercuric chloride treatment activates transcription of the heme oxygenase-1 (HO-1) gene in a dose dependent manner, further indicating an oxidative stress response. Thus, mercury-induced stress may transform innocuous astrocytes into potentially lethal sources of cytotoxic oxygen free radicals. PMID- 9863767 TI - Differential effects of physostigmine and organophosphates on nicotinic receptors in neuronal cells of different species. AB - The effects of the carbamate physostigmine and of the organophosphates (OPs) parathion, paraoxon and phenyl saligenin cyclic phosphate (PSP) were examined on different subtypes of neuronal nicotinic acetylcholine receptors (nAChR). Stimulation with 1 mM ACh induced transient nicotinic inward currents in mouse N1E-115 and human SH-SY5Y neuroblastoma and in locust thoracic ganglion cells. All four acetylcholinesterase (AChE) inhibitors reduced the nicotinic currents in a concentration-dependent manner. Parathion is about 50 times more potent in blocking nAChR, compared to its active AChE inhibiting metabolite paraoxon. The relative blocking potency of the different AChE inhibitors was the same in all cell types, and followed the order parathion > physostigmine > PSP > paraoxon. In N1E-115 cells the IC50 values of block amounted to 2 microM, 30 microM, 39 microM and 96 microM for parathion, physostigmine, PSP and paraoxon, respectively. In all cell types, the nicotinic currents were equally blocked by parathion. Human nAChR in SH-SY5Y cells appeared more sensitive to block by physostigmine, PSP and paraoxon, while these AChE inhibitors similarly inhibited nicotinic currents in insect cells and in mouse neuroblastoma cells. The observation that the concentration-dependence of block is different from that of AChE inhibition, indicates a distinct interaction of AChE inhibitors with nAChR. Only in locust cells physostigmine induced a non-desensitizing inward current, that appeared to originate from nAChR activation. Occasionally, the OPs were able to activate slow ionic currents in mouse, but not in human and locust cells. As the OP-induced agonistic activity in mouse cells was not associated with the blocking action, the target site appeared to be distinct from nAChR. These results show that AChE inhibitors block nAChR with different potencies, dependent on the compound and the receptor subtype, and may activate distinct ion currents in neuronal cells of different species origin. PMID- 9863768 TI - Effect of organophosphate administration on the expression of pro opiomelanocortin-derived peptides in motoneurones. AB - Adult male mice were treated with di-isopropylfluorophosphate (DFP), a neurotoxic organophosphate which crosses the blood brain barrier, or ecothiophate (phospholine iodide) which has been reported to be non-neurotoxic. The presence of immunoreactivity for the pro-opiomelanocortin (POMC)-derived peptides beta endorphin and alpha-melanotropin was examined in intramuscular nerves in diaphragm, extensor digitorum longus (EDL) and soleus muscles, and in motoneurones in the cervical and lumbar regions of the spinal cord, up to 5 days after the treatment. The incidence of both immunoreactive intramuscular nerves and immunoreactive ventral horn cells was increased after administration of either DFP or ecothiophate but the extent of the increase and the time course depended on the muscle type or spinal cord region being examined. The effect of the treatment on the expression of POMC mRNA was investigated using in situ hybridization. The proportion of ventral horn cells which expressed the mRNA was significantly increased by 3h after ecothiophate administration. It reached a peak at 24 h but had returned to normal by 48 h. The results indicate that organophosphates can increase the expression of POMC-derived peptides in motoneurones whether the compounds cross the blood-brain barrier freely or not. This effect may be a consequence of their action to inhibit acetylcholinesterase at the neuromuscular junction and may be part of a regenerative response. PMID- 9863769 TI - The effect of pyridostigmine administration on the expression of pro opiomelanocortin-derived peptides in motoneurones. AB - Adult male mice were treated with the reversible acetylcholinesterase inhibitor pyridostigmine bromide and its effect on the expression of beta-endorphin and alpha-melanotropin immunoreactivity in motoneurones was examined in the cervical and lumbar spinal cord. Administration of a single dose of either 0.4 micromoles/kg or 2.0 micromoles/kg body weight caused a significant increase in the incidence of beta-endorphin and alpha-melanotropin-immunoreactive motoneurones at 3h after the injection, in both the cervical and lumbar regions of the spinal cord. The immunoreactivity remained elevated for at least 5 days. There was a significantly higher increase in both beta-endorphin and alpha melanotropin immunoreactive neurones at 3h after the higher dose compared to the lower dose, in both regions of the spinal cord. Repeated administration of a dose of 0.4 micromoles/kg once a day for three weeks caused increases in immunoreactive motoneurones in both regions. In the cervical region the increases were maintained for at least two weeks after the treatment was discontinued but in the lumbar region the levels had returned to normal by one week. A further dose of the drug administered at two weeks after the treatment period caused a significantly greater increase in the lumbar spinal cord than the same dose in untreated mice, indicating that a sensitization of the motoneurones had occurred. The effect of this drug on peptide expression in motoneurones may be secondary to its action to inhibit acetylcholinesterase at the neuromuscular junction. PMID- 9863770 TI - Evaluation of subchronic neurotoxicity of n-butyl acetate vapor. AB - n-Butyl acetate, a common industrial solvent, was selected by the US EPA as a chemical of concern for neurotoxicity as part of the Multisubstance Rule for the Testing of Neurotoxicity. The neurotoxic potential of n-butyl acetate was investigated in Sprague-Dawley rats using a functional observational battery, motor activity, neurohistopathology, and schedule-controlled operant behavior (SCOB) as indicators of neurotoxicity. Animals were exposed to concentrations of 0, 500, 1500, or 3000 ppm of n-butyl acetate for 6 hours per day for 65 exposures over 14 weeks. Functional observational battery and motor activity values for ad libitum-fed male and female rats were measured during Weeks -1, 4, 8, and 13. SCOB testing of food-restricted animals, using a multiple fixed ratio/fixed interval schedule, was conducted daily prior to each exposure to maintain the operant behavior; the data from Weeks -1, 4, 8, and 13 were evaluated for evidence of neurotoxicity. Transient signs of sedation and hypoactivity were observed only during exposure to the 1500 and 3000 ppm concentrations. The only signs of systemic toxicity were reduced body weights for the 3000 ppm ad libitum fed groups and occasionally for the female 1500 ppm ad libitum-fed group. No evidence of neurotoxicity was seen during the functional observational battery examinations. Motor activity for the 3000 ppm male group was significantly (p < or = 0.05) higher than for the control group only during Week 4. No significant differences were observed among groups for Weeks 8 and 13. No significant differences in motor activity values were observed for female rats. No significant differences were seen in operant behavior at any test vapor concentration. Microscopic evaluations of sections from the brain, spinal cord (cervical and lumbar regions), dorsal and ventral spinal roots, dorsal root ganglia, sciatic nerve, and tibial nerve of animals in the control and 3000 ppm groups did not indicate any treatment-related effects. In conclusion, there was no evidence of cumulative neurotoxicity based on the functional observational battery, motor activity, neurohistopathology, and schedule-controlled operant behavior endpoints. The data presented here are relevant to the neurotoxicity risk assessment of n-butanol due to the rapid hydrolysis of n-butyl acetate in vivo. PMID- 9863771 TI - Action of the pyrethroid insecticide cypermethrin on rat brain IIa sodium channels expressed in xenopus oocytes. AB - Pyrethroid insecticides bind to a unique site on voltage-dependent sodium channels and prolong sodium currents, leading to repetitive bursts of action potentials or use-dependent nerve block. To further characterize the site and mode of action of pyrethroids on sodium channels, we injected synthetic mRNA encoding the rat brain IIa sodium channel alpha subunit, either alone or in combination with synthetic mRNA encoding the rat sodium channel beta1 subunit, into oocytes of the frog Xenopus laevis and assessed the actions of the pyrethroid insecticide [1R,cis,alphaS]-cypermethrin on expressed sodium currents by two-electrode voltage clamp. In oocytes expressing only the rat brain IIa alpha subunit, cypermethrin produced a slowly-decaying sodium tail current following a depolarizing pulse. In parallel experiments using oocytes expressing the rat brain IIa alpha subunit in combination with the rat beta1 subunit, cypermethrin produced qualitatively similar tail currents following a depolarizing pulse and also induced a sustained component of the sodium current measured during a step depolarization of the oocyte membrane. The voltage dependence of activation and steady-state inactivation of the cypermethrin dependent sustained current were identical to those of the peak transient sodium current measured in the absence of cypermethrin. Concentration-response curves obtained using normalized tail current amplitude as an index of the extent of sodium channel modification by cypermethrin revealed that coexpression of the rat brain IIa alpha subunit with the rat beta1 subunit increased the apparent affinity of the sodium channel binding site for cypermethrin by more than 20 fold. These results confirm that the pyrethroid binding site is intrinsic to the sodium channel alpha subunit and demonstrate that coexpression of the rat brain IIa alpha subunit with the rat beta1 subunit alters the apparent affinity of this site for pyrethroids. PMID- 9863772 TI - Quinidine prevents paraoxon-induced necrotizing myopathy in rats. AB - Acute organophosphorus anticholinesterase poisoning induces a necrotizing end plate myopathy in rats and patients. Acetylcholine (ACh) excess leads to prolonged synaptic currents and increased influx of cations including calcium through the postsynaptic ACh receptor channels with prolonged muscle membrane depolarization, excess calcium influx into the sarcoplasm, and ultimately muscle fiber necrosis. Quinoline derivatives such as quinidine induce or worsen pre- and postsynaptic disorders of neuromuscular transmission in humans, and are beneficial in patients suffering from a rare congenital myasthenic syndrome called the slow channel congenital myasthenic syndrome. These drugs correct the prolonged opening times of the mutated acetylcholine receptor channels in this myasthenic syndrome. We treated paraoxon-poisoned rats with 4 x 10 or 4 x 50 mg/kg of quinidine and assessed the severity of the necrotizing myopathy in gastrocnemius and diaphragm muscle biopsies. Fasciculations were decreased and the necrotizing myopathy was prevented in most treated rats, with absence of necrotic muscle fibers in most animals in the high-dose group. Survival was not different from untreated poisoned animals. A number of physiological mechanisms, including blocking of presynaptic voltage-gated sodium or calcium channels or inhibition of the postsynaptic ACh receptors channels may have contributed to the attenuation of the myonecrosis. The optimal dose and the drug of choice amongst the clinically available quinoline derivatives remains to be determined. PMID- 9863773 TI - Regionally specific changes in central brain monoamine levels by 2,4 dichlorophenoxyacetic acid in acute treated rats. AB - As reported in our previous studies, "Serotonin Syndrome", rearing and circling behaviors, were the responses of rats treated with the herbicide 2,4 dichlorophenoxyacetic acid (2,4-D) and challenged with amphetamine. An altered serotonergic and dopaminergic system may be responsible for these behaviors. In this work, endogenous levels of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), serotonin (5-HT), and 5-hydroxyindolacetic acid (5-HIAA) were measured in the areas of the mesocortico-limbico and nigrostriatal systems of rats treated with 50 or 100 mg/kg of 2,4-D, but also challenged with amphetamine (5 or 10 mg/kg), in order to determine regional brain contents of biogenic amines and their metabolites and to find a correlation with the behaviors described. The prefrontal cortex (PFc), striatum, nucleus accumbens (NA), midbrain, ventral tegmental area (VTA), substantia nigra (SN) and cerebellum were assayed. The herbicide 2,4-D increased the 5-HT and DA basal levels and these increments were dependent on sex, brain area and 2,4-D doses. The challenge with 5 or 10 mg/kg amphetamine to 2,4-D (50 or 100 mg/kg) treated rats also resulted in an area-, sex- and dose-dependent alterations of the 2,4-D effect on dopaminergic and serotonergic systems. We are demonstrating a correlation between rat behavior altered by the herbicide's treatment and the contents of 5-HT and DA in brain areas, such as PFc, VTA, SN, striatum, NA, midbrain and cerebellum. In order for 2,4-D to produce the neurobehavioral alterations, not only are high levels of 5 HT and DA necessary, but these neurotransmitters must be released. PMID- 9863774 TI - Organophosphate-induced acetylcholinesterase inhibition and embryonic retinal cell necrosis in vivo in the teleost (Oryzias latipes). AB - Recent monitoring of the Sacramento-San Joaquin River system (CA) indicates that levels of the organophosphate pesticide, diazinon, exceed National Academy of Science guidelines and these levels result in toxicity in USEPA acute toxicity tests with Ceriodaphnia dubia. Since organophosphates (OPs) inhibit acetylcholinesterase (AChE), the present study examined the effects of diazinon on the embryonic nervous system of a model teleost, medaka, Oryzias latipes. Preliminary histological screens revealed limited retinal cell necrosis in control embryos with apparent increased necrosis in diazinon-exposed embryos. Subsequently, embryos were exposed to 1.8 x 10(-5), 4.4 x 10(-5), or to 8.8 x 10( 5) M diazinon and replicates were frozen for biochemical analysis or were fixed for histopathological analysis at days 3, 5, and 7 of development. Diazinon exposure significantly inhibited AChE activity within whole embryos and in homogenates of retinas from treated animals. Histological examination of embryos indicated that as the retina underwent differentiation into distinct cell layers, between days 5 and 7, small foci of necrotic cells became apparent within the inner nuclear layer and isolated individual pyknotic cells were observed in the ganglion layer. Quantification of foci of necrotic cells revealed that 8.8 x 10( 5) M diazinon increased number and area of these lesions. Enzyme histochemistry localized AChE activity to regions equivalent to sites of necrosis. Separate exposures of embryos to the OP, diisopropylphosphorofluoridate, produced large foci of necrotic cells at sites equivalent to those seen following diazinon exposure. PMID- 9863775 TI - Field screening of blood lead levels in remote Andean villages. AB - Blood lead (PbB) levels were investigated in chronically lead (Pb) exposed Andean children and adults living in a highly Pb contaminated area of Ecuador where Pb glazing of ceramics is prevalent. A comparative study was made of the PbB levels of Pb-glazing and non-Pb-glazing families living in close proximity, using three PbB analysis techniques. Fifty-one, 50-microl blood samples from children and adults were analyzed in the field by a finger-stick capillary screening technique using the portable ESA LeadCare Blood Lead Testing System (LCS). Venous blood samples of 2-4 ml were collected from the same 51 participants and analyzed in the laboratory by inductively coupled plasma mass spectrometry (ICP-MS) and by graphite furnace atomic absorption spectrometry (AAS). The median PbB levels for the Pb-glazing group as determined by the ICP-MS, AAS and LCS techniques were 37.2 microg/dl (range 11.6-101.0), 32.0 microg/dl (range 8.0-70.0 microg/dl) and 44.0 microg/dl (range 19.0-105.0), respectively. The median PbB levels for the non-Pb-glazing group were 9.2 microg/dl (range 5.0-21.7) with ICP-MS, 9.0 microg/dl (range 4.3-32.0) with AAS, and 11.3 microg/dl (range 7.3-21.1) with LCS. The differences in PbB levels between the Pb glazing and non-Pb glazing groups were statistically significant (p = < .0001) for each PbB analysis method. Correlations between paired samples were: LCS and ICP-MS: r = 0.913, LCS and AAS: r = 0.829, and ICP-MS and AAS: r = 0.905. The results suggest that neighboring Pb glazing and non-Pb glazing families have significantly different PbB levels, and that the portable LCS field technique may be useful for screening and periodic monitoring of relatively low and high PbB levels of persons in remote high altitude Andean areas. PMID- 9863776 TI - Prenatal and lactational exposure to methylmercury affects select parameters of mouse cerebellar development. AB - Animal studies of the neuropathological effects of prenatal methylmercury (MeHg) seldom use regimens that represent environmental exposures. While acute administration of high doses of MeHg to developing rodents can model some of the outcomes MeHg produces in the human cerebellum, their long-term relevance to cerebellar development is unknown. The present study was undertaken to determine the effect of chronic dietary exposure to MeHg. Pregnant mice were exposed throughout gestation to 0.0 or 4.0 ppm methylmercury in their drinking water. Postpartum exposure of pups and lactating dams continued to postnatal day (PND) 30. On PND7, 14, 21, and 30, several morphometric indices of cerebellar cortex development, as well as blood and brain levels of total Hg, were measured in pairs of male and female littermates. No signs of overt toxicity were observed in the dams or offspring. Blood and brain levels of total Hg were highest in the exposed PND7 offspring and fell throughout the sampling period despite continued exposure. In a region of molecular layer in the anterodorsal lobe, MeHg exposure reduced the density of migrating cells in PND7 offspring. Molecular layer widths were reduced in PND30 offspring. In a region of the inferior lobe of PND7 offspring, MeHg exposure reduced external granular layer widths and decreased the density of migrating cells in the molecular layer. However, MeHg did not affect cerebellar cortex development in the central lobe, suggesting a regional sensitivity to chronic, low-level MeHg exposure during development. PMID- 9863777 TI - Effects of lifetime lead exposure on spatial and temporal visual function in monkeys. AB - Detailed characterization of several aspects of visual function was performed in two groups of monkeys (Macaca fascicularis) exposed to lead continuously from birth. One group of four monkeys was dosed from birth onward with 500 microg/kg/day of lead as lead acetate, while another group of six monkeys was dosed with 2000 microg/kg/day. Blood lead levels peaked in the former group at 50 microg/dl early in life; the latter group had peak blood lead concentrations averaging 115 microg/dl during infancy. Blood lead concentrations decreased before one year of age to stable levels of 25 or 35 microg/dl in the low and high dose groups respectively. Spatial and temporal visual function was assessed using a psychophysical procedure at 7-9 years of age. Six age-matched controls were tested concurrently. Two treated monkeys had severe hyperopia: one was assessed on temporal vision only and the other was not assessed. Spatial data from a third monkey were considered suspect and excluded. Six of the nine treated monkeys in which temporal vision was assessed had thresholds below control values at low and/or middle frequencies under high luminance conditions. Low-luminance temporal vision was not affected. There was no evidence of impairment of spatial visual function in lead-exposed monkeys with normal refractive status at either high or low luminance. This study suggests that temporal visual function may be preferentially impaired as a result of lifetime exposure to a moderate body burden of lead, although these results require replication before definitive conclusions may be drawn. PMID- 9863778 TI - Driving under the influence. PMID- 9863779 TI - Treating influenza with zanamivir. PMID- 9863780 TI - Alcohol in the myocardial infarction patient. PMID- 9863781 TI - Advances in treatment of systemic sclerosis. PMID- 9863782 TI - Loss to follow-up of preterm and very preterm babies. PMID- 9863783 TI - Development of clinical guidelines. PMID- 9863784 TI - Randomised trial of efficacy and safety of inhaled zanamivir in treatment of influenza A and B virus infections. The MIST (Management of Influenza in the Southern Hemisphere Trialists) Study Group. AB - BACKGROUND: Influenza affects many people worldwide each year and has many troublesome symptoms. We investigated the efficacy and safety of the inhaled antiviral agent zanamivir as a treatment for influenza A and B infection. METHODS: In a randomised, double-blind, placebo-controlled trial, we recruited 455 patients aged 12 years and older with influenza-like symptoms of 36 h duration or less who lived in Australia, New Zealand, and South Africa. Eligible patients were randomly assigned 10 mg inhaled zanamivir (n=227) or placebo (n=228) twice daily for 5 days. All patients recorded symptoms on diary cards four times daily during treatment and twice daily for 9 days after treatment. We analysed all patients by intention to treat, influenza-positivity, and high risk of developing complications. FINDINGS: Compared with placebo, zanamavir relieved influenza symptoms a median of 1.5 days earlier in the intention-to-treat (p=0.011) and influenza-positive (p=0.004) populations, and 2 days earlier in patients who were febrile at entry [corrected]. In high-risk patients treated with zanamivir, symptoms were alleviated a median of 2.5 days earlier (p=0.048), fewer had complications (p=0.004), and fewer used complication-associated antibiotics (p=0.025) compared with placebo. The adverse event profiles were similar for zanamivir and placebo. INTERPRETATION: Zanamivir was well-tolerated and effective in decreasing the duration and severity of symptoms. Complications were also decreased in high-risk patients but these findings need to be confirmed in future studies due to the limited number of patients [corrected]. PMID- 9863785 TI - Mortality and light to moderate alcohol consumption after myocardial infarction. AB - BACKGROUND: Although heavy alcohol consumption increases total mortality, light to moderate consumption decreases cardiovascular and all-cause mortality in apparently healthy people. Since data are sparse on the relation of light to moderate alcohol intake to mortality in patients with previous myocardial infarction, we did a prospective study of mortality in men. METHOD: Of 90,150 men in the Physicians' Health Study enrollment cohort who provided information on alcohol intake and who had no history of cancer, stroke, or liver disease, 5358 had a previous myocardial infarction. We estimated alcohol consumption by food frequency questionnaire. FINDINGS: During a mean follow-up of 5 years, 920 men died. After adjustment for several potential confounders, moderate alcohol intake was associated with a significant decrease in total mortality (p=0.016). Compared with men who rarely or never drank alcohol, those who drank one to four drinks per month had a relative risk for total mortality of 0.85 (95% CI 0.69-1.05); for two to four drinks per week, the relative risk was 0.72 (0.58-0.89); for one drink per day 0.79 (0.64-9.96); and for two or more drinks per day 0.84 (0.55 1.26). INTERPRETATION: Men with previous myocardial infarction who consume small to moderate amounts of alcohol have a lower total mortality. PMID- 9863786 TI - Prospects for worldwide tuberculosis control under the WHO DOTS strategy. Directly observed short-course therapy. AB - BACKGROUND: WHO advocates the use of directly observed treatment with a short course drug regimen as part of the DOTS strategy, but the potential effect of this strategy worldwide has not been investigated. METHODS: We developed an age structured mathematical model to explore the characteristics of tuberculosis control under DOTS, and to forecast the effect of improved case finding and cure on tuberculosis epidemics for each of the six WHO regions. FINDINGS: In countries where the incidence of tuberculosis is stable and HIV-1 absent, a control programme that reaches the WHO targets of 70% case detection and 85% cure would reduce the incidence rate by 11% (range 8-12) per year and the death rate by 12% (9-13) per year. If tuberculosis has been in decline for some years, the same case detection and cure rates would have a smaller effect on incidence. DOTS saves a greater proportion of deaths than cases, and this difference is bigger in the presence of HIV-1. HIV-1 epidemics cause an increase in tuberculosis incidence, but do not substantially reduce the preventable proportion of cases and deaths. Without greater effort to control tuberculosis, the annual incidence of the disease is expected to increase by 41% (21-61) between 1998 and 2020 (from 7.4 million to 10.6 million cases per year). Achievement of WHO targets by 2010 would prevent 23% (15-30) or 48 million cases by 2020. INTERPRETATION: The potential effect of chemotherapy (delivered as DOTS) on tuberculosis is greater in many developing countries now than it was in developed countries 50 years ago. To exploit this potential, case detection and cure rates urgently need to be improved in the main endemic areas. PMID- 9863787 TI - Hereditary cerebral cavernous angiomas: clinical and genetic features in 57 French families. Societe Francaise de Neurochirurgie. AB - BACKGROUND: Cavernous angiomas, which are vascular malformations mostly located in the central nervous system, may be inherited as an autosomal dominant disorder known as familial cerebral cavernoma (FCC). FCC has been studied in Hispanoamerican families, in which a strong founder effect was shown. We studied the families of 57 non-Hispanic patients with cavernous angiomas. METHODS: All 28 neurosurgery centres in France collaborated in the study. Inclusion criteria were: families of index patients known to have at least one clinically affected relative, and families of index patients with multiple cavernous angiomas who initially presented as sporadic cases. Clinical and cerebral magnetic resonance imaging (MRI) investigations were done in all patients and in other at-risk individuals who consented to take part. FINDINGS: On MRI, 16 of 22 sporadic index patients had relatives with cavernous angiomas. 51 multiple-case families, including 100 patients with symptoms and 164 symptom-free individuals had MRI lesions. Most FCC patients had multiple lesions and there was a strong correlation between number of lesions and age (p<0.01). The sensitivity of gradient-echo sequences was higher than that of standard MRI for detection of small cavernous angiomas. Pattern of inheritance was autosomal dominant, with incomplete clinical penetrance. The occurrence of de-novo mutations was strongly suggested in some families. INTERPRETATION: Neuroimaging penetrance of FCC is much higher than clinical penetrance. 75% of sporadic cases with multiple lesions are in fact familial cases. The proportion of patients developing clinical symptoms is higher in the hereditary form than in the sporadic form of the disorder. PMID- 9863788 TI - Fetal DNA in skin of polymorphic eruptions of pregnancy. AB - BACKGROUND: Polymorphic eruptions of pregnancy (PEP) are common cutaneous disorders of unknown origin that occur usually after week 34 of gestation. Since pregnancy is associated with peripheral-blood chimerism, particularly during the third trimester, we studied the role of fetal cells in the development of the skin lesions. METHODS: We studied samples of skin from ten women with PEP who were carrying male fetuses and 26 women with normal skin or non-PEP skin disorders (13 carrying male and 13 carrying female fetuses). Epidermis and dermis were dissected from the samples, and the DNA was extracted. PCR with primers specific for the SRY gene was used to detect male DNA. FINDINGS: Male DNA was detected in dermis or epidermis from skin lesions of six of the ten women with PEP. No male DNA was detected in any of the 26 women without PEP. INTERPRETATION: Fetal cells can migrate to skin during gestation, where they seem to be associated with the development of cutaneous disorders of pregnancy. PMID- 9863789 TI - Brown-Sequard at a theme park. PMID- 9863790 TI - Unrecognised iron deficiency in critical illness. PMID- 9863791 TI - Low-molecular-weight heparin for immediate management of thromboembolic disease in pregnancy. PMID- 9863792 TI - Maternal plasma fetal DNA as a marker for preterm labour. PMID- 9863793 TI - Expression of CD38 on CD8 T cells predicts maintenance of high viraemia in HAART treated HIV-1-infected children. Highly active antiretroviral therapy. PMID- 9863794 TI - HIV-1 peripheral neuropathy and combination antiretroviral therapy. PMID- 9863795 TI - Delayed occurrence of Graves' disease after immune restoration with HAART. Highly active antiretroviral therapy. PMID- 9863796 TI - Inadequate reporting of controlled trials as short reports. PMID- 9863797 TI - Improvements in childhood mortality in the Gambia. PMID- 9863798 TI - Vasculitis with acute kidney failure and torasemide. PMID- 9863800 TI - Susan Greenfield: making the right connections. PMID- 9863799 TI - Trends in geographical inequalities in provision of general practitioners in England and Wales. PMID- 9863801 TI - National Institutes of Health PhD plan raises objections. PMID- 9863802 TI - Antiretroviral treatment in 1998. PMID- 9863803 TI - North-South twinning in paediatric haemato-oncology: the La Mascota programme, Nicaragua. AB - We describe the La Mascota twinning programme between La Mascota paediatric hospital in Managua, Nicaragua, and hospitals in Monza and Milan, Italy, and Bellinzona, Switzerland. The programme was based on the belief that an attempt to reduce the gap in mortality from cancer in childhood between developed and less developed countries should become an integral part of the care and research activity of a haemato-oncological department of a developed country and not simply an exercise in solidarity. This programme for acute lymphoblastic leukaemia shows that intellectual, organisational, and financial resources can be generated by a twinning programme. What is vital for such programmes is a long term commitment to a comprehensive and holistic strategy that incorporates supply of drugs, training and supervision of health professionals, and the care of the children and of their parents. PMID- 9863804 TI - Choriodecidual inflammation: a potentially preventable cause of perinatal HIV-1 transmission? AB - The obstetric risk factors for perinatal transmission of HIV-1 include preterm birth, prolonged rupture of the chorioamniotic membranes, and clinical and histological bacterial chorioamnionitis. A chronic chorioamnionitis precedes many cases of preterm labour and spontaneous rupture of membranes, whereas an acute chorioamnionitis is more common after rupture of the membranes at term. Amniotic fluid cytokines are raised in the presence of term and preterm intrauterine bacterial infections, and various cytokines seem able to attract HIV-1-infected leucocytes into the amniotic cavity and to increase replication of HIV-1. We postulate that the association of preterm birth and prolonged rupture of membranes with perinatal transmission of HIV-1 may result from an associated chronic or acute bacterial chorioamnionitis marked by the migration of HIV-1 infected maternal leucocytes into the amniotic cavity. Antibiotic treatment could prevent this sequence of events. PMID- 9863805 TI - The good old days. PMID- 9863806 TI - The UK Prospective Diabetes Study. PMID- 9863807 TI - The UK Prospective Diabetes Study. PMID- 9863808 TI - The UK Prospective Diabetes Study. PMID- 9863809 TI - The UK Prospective Diabetes Study. PMID- 9863810 TI - The UK Prospective Diabetes Study. PMID- 9863811 TI - The UK Prospective Diabetes Study. PMID- 9863812 TI - Anthelmintic treatment for infestation with Trichuris trichiura. PMID- 9863813 TI - Early treatment of HIV-1 infection. PMID- 9863814 TI - Early treatment of HIV-1 infection. PMID- 9863815 TI - Early treatment of HIV-1 infection. PMID- 9863816 TI - Breastfeeding and maternal employment. PMID- 9863817 TI - Acute myocardial infarction associated with sildenafil. PMID- 9863818 TI - Acute myocardial infarction associated with sildenafil. PMID- 9863819 TI - ACE inhibitors and pneumonia in elderly people. PMID- 9863820 TI - Classification of inflammatory arthritis. PMID- 9863821 TI - Classification of inflammatory arthritis. PMID- 9863822 TI - Emergency contraception with levonorgestrel or the Yuzpe regimen. Task Force on Postovulatory Methods of Fertility Regulation. PMID- 9863823 TI - Emergency contraception with levonorgestrel or the Yuzpe regimen. PMID- 9863824 TI - Crisis after angiography: time for MIBG. PMID- 9863825 TI - Parathyroid hormone and corticosteroid-induced osteoporosis. PMID- 9863826 TI - Use of medical information by UK insurers. PMID- 9863827 TI - Use of medical information by UK insurers. PMID- 9863828 TI - South African health-budget cuts hit premature babies. PMID- 9863829 TI - Death from "non-lethal" firearm. PMID- 9863830 TI - Defeating or preventing stigma of mental illness? PMID- 9863831 TI - The vibrating pager. PMID- 9863832 TI - The Nobel chronicles. 1930: Karl Landsteiner (1868-1943). PMID- 9863833 TI - Sketches from The Lancet. The great war. PMID- 9863834 TI - European neurologists focus on aging. PMID- 9863835 TI - Migraine as a woman's issue--will research and new treatments help? PMID- 9863836 TI - The doctor is still in: Freud exhibit starts its tour at Library of Congress. PMID- 9863837 TI - From the Centers for Disease Control and Prevention. Response to increases in cigarette prices by race/ethnicity, income, and age groups--United States, 1976 1993. PMID- 9863838 TI - From the Centers for Disease Control and Prevention. Progress toward poliomyelitis eradication--West Africa, 1997-September 1998. PMID- 9863839 TI - Natriuretic peptides--relevance in cardiovascular disease. PMID- 9863840 TI - A piece of my mind. First attending rounds. PMID- 9863841 TI - Ethnic and socioeconomic factors as determinants of health status. PMID- 9863842 TI - Triglycerides and small, dense low-density lipoprotein. PMID- 9863843 TI - Excessive weight gain and effects on lipids with intensive therapy of type 1 diabetes. PMID- 9863844 TI - Excessive weight gain and effects on lipids with intensive therapy of type 1 diabetes. PMID- 9863845 TI - Temporal artery biopsy to diagnose temporal arteritis. PMID- 9863846 TI - Cost-effectiveness analyses of statistically ineffective treatments. PMID- 9863847 TI - Preventing low back pain in industry. PMID- 9863848 TI - Preventing low back pain in industry. PMID- 9863849 TI - Hot butter and cheese dispensers at the movie theater: a real sizzler? PMID- 9863850 TI - Behavioral vs drug treatment for urge urinary incontinence in older women: a randomized controlled trial. AB - CONTEXT: Urinary incontinence is a common condition caused by many factors with several treatment options. OBJECTIVE: To compare the effectiveness of biofeedback assisted behavioral treatment with drug treatment and a placebo control condition for the treatment of urge and mixed urinary incontinence in older community dwelling women. DESIGN: Randomized placebo-controlled trial conducted from 1989 to 1995. SETTING: University-based outpatient geriatric medicine clinic. PATIENTS: A volunteer sample of 197 women aged 55 to 92 years with urge urinary incontinence or mixed incontinence with urge as the predominant pattern. Subjects had to have urodynamic evidence of bladder dysfunction, be ambulatory, and not have dementia. INTERVENTION: Subjects were randomized to 4 sessions (8 weeks) of biofeedback-assisted behavioral treatment, drug treatment (with oxybutynin chloride, possible range of doses, 2.5 mg daily to 5.0 mg 3 times daily), or a placebo control condition. MAIN OUTCOME MEASURES: Reduction in the frequency of incontinent episodes as determined by bladder diaries, and patients' perceptions of improvement and their comfort and satisfaction with treatment. RESULTS: For all 3 treatment groups, reduction of incontinence was most pronounced early in treatment and progressed more gradually thereafter. Behavioral treatment, which yielded a mean 80.7% reduction of incontinence episodes, was significantly more effective than drug treatment (mean 68.5% reduction; P=.04) and both were more effective than the placebo control condition (mean 39.4% reduction; P<.001 and P=.009, respectively). Patient-perceived improvement was greatest for behavioral treatment (74.1% "much better" vs 50.9% and 26.9% for drug treatment and placebo, respectively). Only 14.0% of patients receiving behavioral treatment wanted to change to another treatment vs 75.5% in each of the other groups. CONCLUSION: Behavioral treatment is a safe and effective conservative intervention that should be made more readily available to patients as a first-line treatment for urge and mixed incontinence. PMID- 9863851 TI - Intensive lifestyle changes for reversal of coronary heart disease. AB - CONTEXT: The Lifestyle Heart Trial demonstrated that intensive lifestyle changes may lead to regression of coronary atherosclerosis after 1 year. OBJECTIVES: To determine the feasibility of patients to sustain intensive lifestyle changes for a total of 5 years and the effects of these lifestyle changes (without lipid lowering drugs) on coronary heart disease. DESIGN: Randomized controlled trial conducted from 1986 to 1992 using a randomized invitational design. PATIENTS: Forty-eight patients with moderate to severe coronary heart disease were randomized to an intensive lifestyle change group or to a usual-care control group, and 35 completed the 5-year follow-up quantitative coronary arteriography. SETTING: Two tertiary care university medical centers. INTERVENTION: Intensive lifestyle changes (10% fat whole foods vegetarian diet, aerobic exercise, stress management training, smoking cessation, group psychosocial support) for 5 years. MAIN OUTCOME MEASURES: Adherence to intensive lifestyle changes, changes in coronary artery percent diameter stenosis, and cardiac events. RESULTS: Experimental group patients (20 [71%] of 28 patients completed 5-year follow-up) made and maintained comprehensive lifestyle changes for 5 years, whereas control group patients (15 [75%] of 20 patients completed 5-year follow-up) made more moderate changes. In the experimental group, the average percent diameter stenosis at baseline decreased 1.75 absolute percentage points after 1 year (a 4.5% relative improvement) and by 3.1 absolute percentage points after 5 years (a 7.9% relative improvement). In contrast, the average percent diameter stenosis in the control group increased by 2.3 percentage points after 1 year (a 5.4% relative worsening) and by 11.8 percentage points after 5 years (a 27.7% relative worsening) (P=.001 between groups. Twenty-five cardiac events occurred in 28 experimental group patients vs 45 events in 20 control group patients during the 5-year follow-up (risk ratio for any event for the control group, 2.47 [95% confidence interval, 1.48-4.20]). CONCLUSIONS: More regression of coronary atherosclerosis occurred after 5 years than after 1 year in the experimental group. In contrast, in the control group, coronary atherosclerosis continued to progress and more than twice as many cardiac events occurred. PMID- 9863852 TI - Tuberculin skin test screening practices among US colleges and universities. AB - CONTEXT: Concern about transmission of Mycobacterium tuberculosis on college campuses has prompted some schools to institute tuberculin skin test screening of students, but this screening has never been evaluated. OBJECTIVE: To describe tuberculin skin test screening practices and results of screening in colleges and universities in the United States. DESIGN AND SETTING: Self-administered mail and telephone questionnaire in November and December 1995 to a stratified random sample of US 2-year and 4-year colleges and universities. MAIN OUTCOME MEASURES: Type of tuberculin screening required; types of schools requiring screening; number and rate of students with positive skin test results and/or diagnosed as having tuberculosis. RESULTS: Of the 3148 US colleges and universities, 624 (78%) of 796 schools surveyed responded. Overall, 378 schools (61%) required tuberculin screening; it was required for all new students (US residents and international students) in 161 (26%) of 624 schools, all new international students but not new US residents in 53 (8%), and students in specific academic programs in 294 (47%). Required screening was more likely in 4-year vs 2-year schools, schools that belonged to the American College Health Association vs nonmember schools, schools with immunization requirements vs schools without, and schools with a student health clinic vs those without (P<.001 for all). Public and private schools were equally likely to require screening (64% vs 62%; P=.21). In the 378 schools with screening requirements, tine or multiple puncture tests were accepted in 95 (25%); test results were recorded in millimeters of induration in 95 (25%); and 100 (27%) reported collecting results in a centralized registry or database. Of the 168 (27%) of 624 schools accepting only Mantoux skin tests and reporting results for school years 1992-1993 through 1995-1996, 3.1% of the 348 368 students screened had positive skin test results (median percentage positive, 0.8%). International students had a significantly higher case rate for active tuberculosis than US residents (35.2 vs 1.1 per 100000 students screened). CONCLUSIONS: Widespread tuberculin screening of students yielded a low prevalence of skin test reactors and few tuberculosis cases. To optimize the use of limited public health resources, tuberculin screening should target students at high risk for infection. PMID- 9863853 TI - Total parenteral nutrition in the critically ill patient: a meta-analysis. AB - CONTEXT: Nutritional support has become a standard of care for hospitalized patients, but whether total parenteral nutrition (TPN) affects morbidity and mortality is unclear. OBJECTIVE: To examine the relationship between TPN and complication and mortality rates in critically ill patients. DATA SOURCES: Computerized search of published research on MEDLINE from 1980 to 1998, personal files, and review of relevant reference lists. STUDY SELECTION: We reviewed 210 titles, abstracts, and papers. Primary studies were included if they were randomized clinical trials of critically ill or surgical patients that evaluated the effect of TPN (compared with standard care) on complication and mortality rates. We excluded studies comparing TPN with enteral nutrition. DATA EXTRACTION: Relevant data were abstracted on the methodology and outcomes of primary studies. Data were abstracted in duplicate, independently. DATA SYNTHESIS: There were 26 randomized trials of 2211 patients comparing the use of TPN with standard care (usual oral diet plus intravenous dextrose) in surgical and critically ill patients. When the results of these trials were aggregated, TPN had no effect on mortality (risk ratio [RR], 1.03; 95% confidence interval [CI], 0.81-1.31). Patients who received TPN tended to have a lower complication rate, but this result was not statistically significant (RR, 0.84; 95% CI, 0.64-1.09). We examined several a priori hypotheses and found that studies including only malnourished patients were associated with lower complication rates but no difference in mortality when compared with studies of nonmalnourished patients. Studies published since 1989 and studies with a higher methods score showed no treatment effect, while studies published in 1988 or before and studies with a lower methods score demonstrated a significant treatment effect. Complication rates were lower in studies that did not use lipids; however, there was no difference in mortality rates between studies that did not use lipids and those studies that did. Studies limited to critically ill patients demonstrated a significant increase in complication and mortality rates compared with studies of surgical patients. CONCLUSIONS: Total parenteral nutrition does not influence the overall mortality rate of surgical or critically ill patients. It may reduce the complication rate, especially in malnourished patients, but study results are influenced by patient population, use of lipids, methodological quality, and year of publication. PMID- 9863854 TI - Techniques to improve physicians' use of diagnostic tests: a new conceptual framework. AB - OBJECTIVES: To review the published literature on interventions aimed at improving physicians' testing practices and propose methodologic standards for these studies and to review selected studies using the PRECEDE framework, a behavioral model that helps categorize interventions based on which behavioral factors are being affected. DATA SOURCES: MEDLINE, EMBASE, and HEALTHStar databases were searched for the years 1966 to January 1, 1998, for English language articles pertaining to diagnostic testing behavior; bibliographies were scanned to identify articles of potential interest; and researchers in health services, health behavior, and behavior modification were contacted for proprietary and other unpublished articles. STUDY SELECTION: A total of 102 articles were identified that described the results of interventions aimed at changing physicians' testing practices. We included the 49 studies that compared diagnostic testing practices in intervention and control groups. DATA EXTRACTION: Two investigators independently reviewed each article in a blinded fashion using a standard data collection form to obtain a methodologic score and to abstract the key elements of each intervention. DATA SYNTHESIS: On a 38-point methodologic criteria scale, the mean +/- SD score was 13+/-4.4. The desired behavior change was reported in the intervention group in 37 (76%) of 49 studies. Twenty-four (86%) of 28 interventions targeted at many behavioral factors were successful, while 13 (62%) of 21 studies aimed at a single behavioral factor were successful (P=.12). CONCLUSIONS: A majority of interventions to improve physicians' testing practices reported in the literature claimed success, with interventions based on multiple behavioral factors trending toward being more successful. While methodologic flaws hamper drawing strong conclusions from this literature, application of a behavioral framework appears to be useful in explaining interventions that are successful and can facilitate interpretation of intervention results. PMID- 9863855 TI - Effect of population-based interventions on laboratory utilization: a time-series analysis. AB - CONTEXT: Previous studies have identified methods of decreasing laboratory utilization. However, most were hospital-based, relatively small, single centered, or of limited duration. OBJECTIVE: To determine the effect of 3 population-based interventions (physician guidelines, laboratory requisition form modification, and changes to funding policy) on laboratory utilization in Ontario. DESIGN: Interventional time-series analysis in which data analysis was based on all claims made to the Ontario Health Insurance Program between July 1, 1991, and April 1997 for laboratory tests affected by the interventions. SETTING: All clinical laboratories (not based in hospitals) in Ontario. INTERVENTIONS: Physician guidelines, modification of laboratory requisition form, and changes in funding policy for the use of the erythrocyte sedimentation rate test (ESR), microscopic urinalysis, tests for renal function, iron stores, serum urea, and serum iron determinations, and tests for thyroid dysfunction (total thyroxine and thyroid-stimulating hormone [TSH]). MAIN OUTCOME MEASURES: Change from 1991 to 1997 in utilization rates of ESR, microscopic urinalysis, serum urea and iron determinations, and tests for total thyroxine and TSH. RESULTS: Age- and sex standardized rates for laboratory tests unaffected by the interventions were stable during the study period. Utilization of ESR and urea determination decreased by 58% (P<.001) and 57% (P<.001), respectively, after they were removed from the requisition form and guidelines discouraging their use were disseminated. Rates for urinalyses without microscopy increased by 1700% (P<.001), while microscopic urinalysis decreased by 14% (P<.001), after a policy change eliminated microscopic urinalysis from routine urinalysis. Rates of iron determination declined by 80% (P<.001) and ferritin rates increased by 34% (P= .05) when policy changes eliminated iron testing when ordered with ferritin and guidelines advocating ferritin alone for investigating iron deficiency were disseminated. Utilization of total thyroxine testing declined by 96% (P = .02) when the provincial health plan stopped its funding. When TSH was removed from the laboratory requisition form, a 12% decline (P= .03) in its use was observed. Through April 1997, these interventions saved more than 625000 tests or $210400. CONCLUSIONS: The combination of guideline dissemination, laboratory requisition form modification, and changes to funding policy was associated with significant reductions in laboratory utilization. PMID- 9863856 TI - Improving treatment of urinary incontinence. PMID- 9863857 TI - Reporting of public health hazards or major advances--revision of uniform requirements. PMID- 9863858 TI - Changing physician behavior in ordering diagnostic tests. PMID- 9863859 TI - The AIDS Net: HIV/AIDS resources on the World Wide Web. PMID- 9863860 TI - "Doctoring" doctors and their families. AB - Being selected to provide medical care to other physicians or their family members represents not only a gratifying professional recognition of competence by one's peers but also a challenge. Many personal and psychological factors may influence the medical care of physicians. III physicians may have difficulty with role reversal and "the VIP syndrome," while treating physicians may have to deal with their own anxiety and issues such as confidentiality, privacy, empathy, and intrusion by a physician-relative into the care of medical family members. Based on experience with more than 200 physician-patients and many adult family members of physicians, suggestions are offered for care of these patient groups. PMID- 9863861 TI - JAMA patient page: incontinence. PMID- 9863862 TI - Comparison of the plasma pharmacokinetics and renal clearance of didanosine during once and twice daily dosing in HIV-1 infected individuals. AB - OBJECTIVE: To compare the plasma pharmacokinetics of didanosine during once daily (qd) and twice daily (bid) dosing. DESIGN: Open-label, randomized, cross-over study. METHODS: HIV-1 infected patients who used didanosine were randomized to either a qd or a bid dosing regimen of didanosine. The total daily dose of didanosine was identical in both regimens. Seven days after the start of the study, the pharmacokinetic profile of didanosine in plasma and urine was assessed during an 8-h period. The next day, the patient was switched to the opposite dosing regimen, and after another 7 days, the study was concluded by again assessing the plasma and urine pharmacokinetics of didanosine during 8 h. RESULTS: A total of 19 patients completed the study. The pharmacokinetics of didanosine in plasma (with maximum plasma concentration adjusted for dose) and urine were not significantly different in the qd and bid dosing regimen (P > 0.28 for all parameters). CONCLUSION: We conclude that qd dosing of didanosine leads to a similar exposure in plasma as bid dosing (using the same total daily dose). Since qd dosing may lead to improved compliance of patients to regimens containing didanosine, these results provide a rationale for prescribing didanosine in a qd regimen, and is reassuring when we realize that the drug is being administered in a qd dosing regimen on a large scale in clinical practice. PMID- 9863863 TI - Restored humoral immune response to influenza vaccination in HIV-infected adults treated with highly active antiretroviral therapy. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) effectively suppresses replication of HIV and is accompanied by an increase in CD4+ T lymphocytes. Whether the increase in CD4+ T lymphocytes in the blood is a reflection of a reconstitution of the immune functions is unknown. We investigated the recovery of the humoral immune response during HAART after immunization with T-cell dependent influenza vaccine. METHODS: Forty-one men and three women infected with HIV and treated with HAART, and 15 healthy hospital staff members were immunized with trivalent influenza subunit vaccine. Antibody titres were determined by haemagglutination inhibiting assay in sera obtained before and 30 days after immunization. Lymphocyte subsets were determined in blood samples taken at the time of vaccination. RESULTS: In all HIV-infected individuals, treatment with HAART caused a median reduction of 2.3 log10 in HIV-1 load. The median increase of CD4+ T lymphocytes after initiation of HAART was 170 x 10(6)/l. The antibody response to influenza antigens was proportional to the number of memory CD4+ T lymphocytes in the blood at the time of vaccination. When a group of patients and healthy controls with approximately similar CD4+ T-lymphocyte counts were considered, the antibody titres after vaccination for influenza strain H1N1 and influenza B did not differ between patients and controls (P=0.12). Vaccination of patients with a CD4+ T-lymphocyte count of < 200 x 10(6)/l (mean 85 x 10(6)/l) before the start of HAART and with a mean of 282 x 10(6)/l CD4+ T lymphocytes at the time of vaccination as a result of HAART, demonstrated a substantial antibody response whereas patients with a CD4+ T lymphocyte count of < 200 x 10(6)/l (mean 56 x 10(6)/l) not treated with HAART (historical controls), and vaccinated with a similar influenza vaccine, failed to induce an antibody response. CONCLUSION: The present findings demonstrate a recovery of the humoral immune response to influenza antigens in HIV-infected individuals treated with HAART. This indicates that functional improvement of antigen specific CD4+ T helper cell reponses occurs. PMID- 9863865 TI - Complications of surgery in HIV-infected patients. PMID- 9863864 TI - Randomized, controlled phase II trial of subcutaneous interleukin-2 in combination with highly active antiretroviral therapy (HAART) in HIV patients. AB - OBJECTIVE: To determine the immunological, virological and clinical effects of subcutaneous IL-2 in 44 HIV-patients in conjunction with pre-existing tri-therapy (zidovudine, 3TC, saquinavir). DESIGN: Partially randomized, controlled, prospective trial. SETTING: Single center study at tertiary care center. PATIENTS: Sixty four patients (CD4 count 200-500 x 10(6)/l). INTERVENTION: Fourty four patients were randomized to receive 5-day cycles of IL-2 (9 Mio IU/d) every 6 weeks (Group A) or whenever the CD4 cell count dropped below the 1.25-fold of baseline (Group B), whereas 20 control patients received the same HAART without IL-2. OUTCOME MEASURES: The optimal individual treatment interval and the immunological and virological effects of subcutaneously administered IL-2 were analysed. Importantly, the level of cellular in vivo immunity and the frequency of dermatological marker diseases and infectious complications were assessed. RESULTS: IL-2 was well tolerated although fever, influenza-like symptoms and indurated injection sites were commonly encountered. After 1 year of IL-2, there was a median increase of more than 100 x 10(6)/l CD4 cells in both IL-2 groups in contrast to the controls (P < 0.01, 0.01 and not significant). The median HIV load did not increase either in plasma or in lymph nodes. Lymphocyte activation decreased as assessed by MHC class II (P < 0.001), CD25 (P < 0.001) and CD38 expression (P < 0.005). Although delayed type hypersensitivity against common recall antigens increased in both IL-2 groups, it did not reach statistical significance. However, it is of note, that in 7 of 11 (63.6%) patients delayed type hypersensitivity against recombinant HIV antigens improved significantly. Whereas there was no opportunistic infection in either IL-2 group, three cases of Kaposi's sarcoma occurred in the controls. Dermatological indicator diseases (thrush, condyloma, herpes simplex) were found to occur more frequently in the control group. CONCLUSIONS: Subcutaneous IL-2 in addition to HAART was safe and led to sustained qualitative and quantitative immunological improvements in the majority of patients. Individualisation of therapy intervals further improved the efficacy and tolerance of IL-2. PMID- 9863866 TI - Levels of HIV-infected peripheral blood cells remain stable throughout the natural history of HIV-1 infection. Swiss HIV Cohort Study. AB - OBJECTIVE: To clarify the relationship between the number of provirus-bearing peripheral blood mononuclear cells (PBMC) and HIV-1 disease progression during the natural history of infection. DESIGN: Twenty-four HIV-1-infected subjects with known seroconversion dates and long-term follow-up were retrospectively identified using the Swiss HIV Cohort Database. PBMC specimens from this cohort were retrieved from storage for analysis. METHODS: Infected PBMC equivalents were determined by HIV-1 DNA quantitative competitive (QC)-PCR. The results were analysed with respect to HIV-1 disease stage and compared with a mathematical model of long-term HIV-1 disease progression. RESULTS: PBMC HIV-1 DNA did not correlate with major indices of disease progression, including time following primary infection, time before reaching a CD4 cell count less than 200 x 10(6)/l, and time before death. The number of PBMC harbouring HIV-1 provirus was relatively constant throughout the clinical stages of HIV-1 infection, consistent with simulated data from a mathematical model of long-term HIV-1 infection. We also showed that a biased interpretation of the QC-PCR data may arise when the values are expressed as HIV-1 DNA copies per PBMC or per CD4 cell. CONCLUSIONS: This analysis suggests that levels of provirus-bearing PBMC remain constant during the natural course of HIV-1 infection, whereas plasma virus load typically increases logarithmically during the same period. The hypothesis that plasma virus levels are directly related to the number of infected cells may deserve reconsideration. PMID- 9863867 TI - Live attenuated simian immunodeficiency virus (SIV)mac in macaques can induce protection against mucosal infection with SIVsm. AB - OBJECTIVE: To investigate whether vaccination of macaques with attenuated simian immunodeficiency virus (SIV)macC8 could induce long-term protective immunity against rectal exposure to SIVsm and intravenous exposure to the more divergent HIV-2. DESIGN AND METHODS: Eight months after vaccination with live attenuated SIVmacC8, four cynomolgus monkeys were challenged with SIVsm intrarectally and another four vaccinated monkeys were challenged with HIV-2 intravenously. Sixteen months after SIVmacC8 vaccination, another two monkeys were challenged with SIVsm across the rectal mucosa. Two vaccinees shown to be protected against SIVsm were rechallenged 8 months after the first challenge. Ten naive animals were used as controls. Serum antigenaemia, virus isolation, antibody responses, cell-mediated immunity and CD4+ and CD8+ T-cell subpopulations were monitored. PCR-based assays were used to distinguish between virus populations. RESULTS: At the time of challenge, eight out of 10 vaccinees were PCR-positive for SIVmacC8 DNA but no virus could be isolated from peripheral blood mononuclear cells. After SIVsm challenge, three out of six vaccinees were repeatedly SIVsm PCR-negative. In one of the three infected monkeys, the challenge virus was initially suppressed but the monkey ultimately developed AIDS after increased replication of the pathogenic virus. Rechallenged monkeys remained protected. All HIV-2-challenged vaccinees became superinfected. All controls became infected with either SIVsm or HIV-2. At the time of challenge the vaccinees had neutralizing antibodies to SIVmac but no demonstrable cross-neutralizing antibodies to SIVsm or HIV-2. Titres of antigen-binding or neutralizing antibodies did not correlate with protection. Cytotoxic T-cell responses to SIV Gag/Pol and virus-specific T-cell proliferative responses were low. CONCLUSION: The live attenuated SIVmacC8 vaccine was able to induce long-term protection against heterologous intrarectal SIVsm challenge in a proportion of macaques but not against the more divergent HIV-2, which was given intravenously. PMID- 9863868 TI - The estimated impact of the CCR-5 delta32 gene deletion on HIV disease progression varies with study design. Oslo HIV Cohort Study Group. AB - OBJECTIVES: To study the impact of the genotype CCR-5 wild-type +/A32 on the progression rate to AIDS and death, and to discuss sources of bias according to study design. METHODS: A prospective study of 310 HIV-positive subjects with follow-up time from study entry (prevalent cohort), and a prospective study of 105 HIV-positive subjects with well-defined time of HIV seroconversion, with follow-up time from the retrospectively assessed date of HIV seroconversion (retrospective incident cohort). RESULTS: Slower progression to AIDS among subjects with CCR-5 +/delta32 than those with CCR-5 +/+ genotype was estimated in the prevalent cohort (P=0.07, log-rank test). Slower progression to death from any cause was also estimated for subjects with CCR-5 +/delta32 (P < 0.05, log rank test). No differences in survival after AIDS diagnosis were seen (P=0.89, log-rank test). No differences in the progression rate to AIDS (P=0.82, log-rank test) or death (P=0.78, log-rank test) were estimated in the retrospective incident cohort. CONCLUSIONS: The varying estimates of the impact of CCR-5 genotype on progression to AIDS in this and other studies, may be real and reflect differences in the dependence of HIV on the CCR-5 receptor, or may be due to systematic errors caused by study design. Several methodological difficulties occur when the factor studied, such as CCR-5 genotype, is associated both with the risk of being HIV-infected and the progression of disease. PMID- 9863869 TI - Presence of the variant mannose-binding lectin alleles associated with slower progression to AIDS. Amsterdam Cohort Study. AB - OBJECTIVE: To examine the association between mannose-binding lectin (MBL) polymorphism and progression to AIDS and death in HIV-1 infection. DESIGN AND METHODS: In 131 HIV-1-infected homosexual seroconverters, survival analyses were performed to determine both the association between MBL genotype and time from HIV-1 seroconversion to AIDS and death, and time from AIDS to death. RESULTS: Of the 131 seroconverters, of whom 61 developed AIDS, 76 were typed as homozygous wild-type and 55 as carriers of variant alleles (52 heterozygous and three homozygous variant alleles). A Survival analyses suggested that HIV-1-infected men with the variant alleles progressed somewhat slower to AIDS [relative hazard (RH), 0.62; 95% confidence interval (CI), 0.36-1.10] and death (RH, 0.73; 95% CI, 0.42-1.25). Interestingly, CD4+ T-cell count determined at the moment of AIDS was found to be significantly lower among persons with the mutation (97 x 10(6)/l versus 204 x 10(6)/l; P=0.03). Furthermore, when AIDS-free times before the diagnosis of an opportunistic infection were compared with those preceding a diagnosis of Kaposi's sarcoma, Kaposi's sarcoma diagnosis was more postponed than that of an opportunistic infection (RH, 0.21; 95% CI, 0.05-0.95; versus RH, 0.67; 95% CI, 0.35-1.27). CONCLUSION: Indications for a weak pre-AIDS protective effect of variant MBL alleles were demonstrated. PMID- 9863870 TI - Selective vertical transmission of HIV-1 antiretroviral resistance mutations. AB - OBJECTIVE: To examine the patterns of vertical transmission of zidovudine (ZDV) resistance mutations. DESIGN: HIV-1 reverse transcriptase codons 10-250 were sequenced from 24 pairs of ZDV-exposed women and their HIV-infected infants as part of the Women and Infants Transmission Study. METHODS: Viral RNA was extracted from tissue culture supernatants and sequenced using fluorescent dye primer chemistry and an automated sequencer. RESULTS: For 17 of these pairs, maternal and infant sequences were identical to one another and lacking known ZDV resistance mutations. The remaining seven maternal sequences contained known mutations associated with ZDV resistance at reverse transcriptase codons 70, 210, 215 and 219. In each case where the maternal HIV isolate showed a pure mutant species, the infant sequence was identical. When the maternal sequence showed the presence of a sequence mixture at codon 70 or 219, the infant's virus showed only wild-type sequence even when the ZDV-resistant mutant was quantitatively dominant in the mother. The single maternal HIV isolate showing mixed sequence at codon positions 210 and 215 transmitted an unmixed mutant to the infant at both positions. When maternal mixtures were present at sites not associated with ZDV resistance, only the dominant species appeared in the infant. CONCLUSIONS: When maternal HIV isolates contained mixed wild-type and ZDV-resistant subpopulations, only a single component of the mixture could be detected in the infected infants. Resistance mutants without the codon 215 mutation were not transmitted from mixtures, even when the mutants formed the majority of circulating maternal virus. In perinatal HIV transmission, specific ZDV-resistant HIV genotypes circulating in the maternal virus pool may influence whether infection in the infant will be established by a wild-type or ZDV-resistant HIV strain. PMID- 9863871 TI - Hepatitis C virus-associated hepatitis following treatment of HIV-infected patients with HIV protease inhibitors: an immune restoration disease? AB - OBJECTIVE: To report observations from case studies on the pathogenic mechanisms underlying the acute hepatitis that sometimes occurs in hepatitis C virus (HCV) and HIV coinfected patients following treatment with potent antiretroviral therapy that includes a HIV protease inhibitor. METHODS: Cases of acute hepatitis were identified from a group of 133 patients enrolled in a retrospective study of pathogen-associated inflammatory disease following the use of potent antiretroviral therapy. Data on serum alanine aminotransferase concentrations, clinical events, HCV antibodies, and liver biopsies were collected from medical records. HCV RNA assays and additional HCV antibody assays were undertaken on stored plasma or sera. RESULTS: Three of the 133 patients (2%) developed symptomatic hepatitis. One was HCV antibody-positive prior to commencing antiretroviral therapy and developed hepatitis subsequent to an episode of Mycobacterium avium complex disease associated with immune restoration. However, the other two patients had previously undiagnosed HCV infection for up to 2 years prior to antiretroviral therapy, with HCV RNA detected but anti-HCV antibody repeatedly undetectable in stored plasma or sera. HCV antibody was only detectable after antiretroviral therapy-induced decrease in plasma HIV RNA and immunological reconstitution. Plasma HCV RNA increased after therapy in one of these patients, but in the other the level was not increased at a time of active hepatitis demonstrated by liver biopsy. CONCLUSIONS: Hepatitis in HCV-HIV coinfected patients following treatment with potent antiretroviral therapy may reflect restoration of anti-HCV immune responses rather than increased HCV replication or a hepatotoxic effect of antiretroviral therapy. PMID- 9863872 TI - Characterizing patterns of drug-taking behavior with a multiple drug regimen in an AIDS clinical trial. AB - OBJECTIVE: To characterize drug-taking behavior using continuous electronic monitoring in an AIDS clinical trial. SETTING: This was a substudy of AIDS Clinical Trials Group (ACTG) protocol 175, a phase II/III study of dideoxynucleoside monotherapy versus combination therapy in asymptomatic HIV positive subjects. Participants were required to comply with regimens containing zidovudine, zalcitabine and didanosine, or matching placebos; the total daily pill count was 16. DESIGN: For participants at two ACTG 175 sites, electronic devices were used to monitor drug-taking behavior of all study medications over a period of approximately 90 days. MEASUREMENTS: Four indices of drug-taking behavior were calculated and their distributions and relationship to the prescribed regimen were examined. RESULTS: Data from 41 subjects were analyzed. Of the prescribed doses of zidovudine, zalcitabine and didanosine, 88, 84 and 82%, respectively, were taken. Of these, 55, 66 and 79%, respectively, were taken at the prescribed dosing frequency. The median percentage of days on which participants failed to take any of the doses was 2-5%. There was a trend towards lower adherence in the combination therapy arms compared with those assigned to receive monotherapy. In this analysis, older patients demonstrated better adherence, although patient characteristics, in general, were poorly predictive of adherence. CONCLUSION: Drug-taking behavior for all three active study medications differed from that prescribed. One result of this erratic adherence was that study participants sustained little antiretroviral effect during more than 25% of the monitoring period. PMID- 9863873 TI - The extent of non-adherence in a large AIDS clinical trial using plasma dideoxynucleoside concentrations as a marker. AB - OBJECTIVES: To assess adherence to study medications in an AIDS clinical trial, to evaluate whether study participants adhered to only one component of a multidrug regimen ('differential adherence'), and to determine whether there was evidence of non-uniform adherence to study medications among treatment groups. SETTING: This was a substudy of AIDS Clinical Trials Group protocol 175, a large, double-blind, randomized study of monotherapy versus combination dideoxynucleoside therapy. Participants were required to adhere to a complex regimen of zidovudine, zalcitabine and didanosine, or their matching placebos. DESIGN: Between October 1992 and January 1994, study sites were selected at random, and a 1-week period was designated during which study participants attending routine clinic visits provided a blood sample and dosing history. Participants were not informed of the purpose of the substudy. MEASUREMENTS: Adherence was assessed using plasma drug concentrations and defined by the presence of detectable drug in a plasma sample obtained within a specified analysis window. RESULTS: Of 722 plasma samples analyzed, approximately 75% contained detectable concentrations of the assigned drugs and 5-14.5% contained no detectable drugs. Approximately 7 and 13% of samples from participants assigned to monotherapy arms contained non-prescribed dideoxynucleosides, and 14 and 19% assigned to combination therapies contained only one drug. CONCLUSIONS: Various non-adherence behaviors were observed, including patterns of underdosing and taking non-prescribed drugs. Non-adherence was moderate but uniform amongst the treatment groups and may have contributed to a marginal reduction in the power of the primary intent-to-treat analysis to detect differences in efficacy amongst the assigned treatments. PMID- 9863874 TI - Changes in CD4+ cell count and the risk of opportunistic infection or death after highly active antiretroviral treatment. Groupe d'Epidemiologie Clinique du SIDA en Aquitaine. AB - OBJECTIVE: To study the relationship between the CD4+ cell response after initiation of protease inhibitors and the occurrence of opportunistic infections and survival. DESIGN: Prospective observational cohort study. METHODS: HIV-1 seropositive subjects followed-up in HIV centres of Bordeaux University Hospital, Southwest France who were prescribed at least one available protease inhibitor between January and December 1996 were included in this analysis. A Cox model estimated the independent effect of baseline covariates and CD4+ cell response, considered as a time-dependent covariate, on the occurrence of new AIDS-defining opportunistic infection, new AIDS-defining events, new AIDS-defining opportunistic infection or death. RESULTS: A total of 556 HIV-positive patients were prescribed at least one protease inhibitor: 34% saquinavir, 52% indinavir, and 14% ritonavir. Median CD4+ cell count at baseline was 95 x 10(6)/l and mean plasma HIV RNA was 5.0 log10 copies/ml. After a median follow-up of 230 days, 65 patients experienced a new episode of opportunistic infection, 79 patients experienced at least one AIDS-defining event, and 24 had died. On average, the increase in CD4+ cell count was 42 x 10(6)/l (SD, 74) after a median of 49 days. In the multivariate analysis of opportunistic infection or death, each 50% higher CD4+ cell count at baseline was associated with a 23% reduction [95% confidence interval (CI), 14-30] of risk. Each 50% increase in CD4+ cell count during follow up was associated with a 9% reduction (95% CI, 2-15) of risk, adjusted for the presence of AIDS prior to protease inhibitor therapy (hazard ratio, 3.76 versus absence of AIDS; P < 0.01) and haemoglobin level (hazard ratio, 0.48 if > 11 g/dl versus <11 g/dl; P < 0.01). CONCLUSION: Our results show, at least indirectly, how protease inhibitors might produce clinical stabilization. This result may be due to improved functionality of CD4+ cells in patients started on protease inhibitors. PMID- 9863875 TI - Catheter complications in AIDS patients treated for cytomegalovirus retinitis. AB - PURPOSE: To describe the complications of central venous catheter use for intravenous therapy of cytomegalovirus (CMV) retinitis in patients with AIDS. METHODS: Retrospective review of 388 patients with AIDS and CMV retinitis treated with intravenous medications through an indwelling catheter. RESULTS: The catheter complication rate was 1.2 complications per person-year (0.33 complications per 100 catheter-days). Current injecting drug use increased the risk of infectious complications [hazard ratio (HR), 1.73; P=0.04] whereas former use did not (HR, 0.96; P=0.88). Subdermal port catheters increased the risk of bacteremia (HR, 1.78; P=0.05). Mortality for the first complication was 5.8%. Forty percent of patients required catheter removal, and 86.8% of these patients required reinsertion of another catheter. CONCLUSIONS: Catheter complications are a substantial problem in patients with CMV retinitis treated with daily intravenous therapy. PMID- 9863876 TI - Improving quality of sexually transmitted disease case management in rural South Africa. AB - OBJECTIVE: To measure quality of sexually transmitted disease (STD) syndromic case management and aspects of health-seeking behaviour at baseline in an intervention trial. SETTING: Ten rural primary care clinics, Hlabisa district, South Africa. DESIGN: Simulated patients (fieldworkers trained to present with STD syndromes) made a total of 44 clinic visits; 49 STD patients were interviewed when exiting clinics; facilities were assessed for availability of necessary equipment and drugs; 10 focus group discussions were held with staff; and STD syndrome surveillance was performed in all 10 clinics. RESULTS: A total of 9% of simulated patients were correctly managed (given correct drugs, plus condoms and partner notification cards), recommended drug treatment was given in only 41% of visits, and appropriate counselling was given in 48% of visits. Among patients leaving the clinic, although 39% waited over an hour to be seen and only 37% were consulted in private, all reported staff attitudes as satisfactory or good. Only six clinics had syndromic management protocols available, three reported intermittent drug shortages, and seven lacked partner notification cards. Focus group discussions revealed good staff knowledge about STD, but showed lack of training in syndromic management and low morale. Surveillance data showed that while 75% of those presenting for care did so within 1 week of symptom onset, 27% had been treated for an STD in the preceding 3 months, and only 6% of those treated were contacts. CONCLUSIONS: Quality of STD case management was poor despite good staff knowledge and availability of most essential resources. An intervention comprising staff training and STD syndrome packets has been designed to improve quality of case management. PMID- 9863877 TI - Attitude of pregnant women towards HIV testing in Abidjan, Cote d'Ivoire and Bobo Dioulasso, Burkina Faso. DITRAME Study Group (ANRS 049 Clinical Trial). Diminution de la Transmission Mere Enfant du VIH. Agence Nationale de Recherches sur le SIDA. AB - OBJECTIVE: To evaluate the attitude of pregnant women towards HIV testing in two cities of West Africa: Abidjan, Cote d'Ivoire and Bobo-Dioulasso, Burkina Faso. METHODS: In the context of a clinical trial to prevent HIV vertical transmission, HIV counselling and testing was offered systematically to women attending antenatal clinics. Informed consent was obtained and test results were given anonymously. Multiple logistic regression was performed to identify factors associated with refusal for testing and failure to return for test results. RESULTS: A total of 9724 pregnant women were interviewed from January 1995 to September 1996. In Abidjan (n=5766) and Bobo-Dioulasso (n=3958), 78 and 92.4% of the women consented to HIV testing, respectively, and 58.4 and 81.8% of them returned for the test results disclosure, respectively. In the two sites, the counsellors themselves and high educational level of the women appeared to be related to refusal of the test, whereas last trimester gestation was associated with failure to return for test results. In Abidjan, foreigners and employees were more likely to refuse testing, and HIV-infected women were three times less likely to return for results than uninfected women. CONCLUSION: Future implementation of interventions to reduce vertical transmission of HIV that require antenatal HIV testing and counselling will have to solve issue of acceptability of HIV testing by pregnant women. PMID- 9863879 TI - Parenteral transmission of Kaposi's sarcoma-associated herpesvirus. PMID- 9863878 TI - Detection of HIV-1 nucleic acid and HIV-1 antibodies in needles and syringes used for non-intravenous injection. AB - BACKGROUND: HIV antibodies and HIV DNA have been detected in needles and syringes that have been used for intravenous injections in HIV-infected persons. During intravenous injection, blood is typically aspirated into the lumen of the syringe. During intramuscular or subcutaneous injection, however, blood is not usually introduced into the syringe. OBJECTIVES: To investigate the presence of HIV antibodies, HIV proviral DNA, HIV RNA, and human DNA in needles and syringes that had been used for intramuscular or subcutaneous injection in persons known to have HIV infection. METHODS: Discarded disposable needles and syringes used by health-care personnel for medically indicated intramuscular or subcutaneous injections of HIV-infected patients were collected. Residual material was extracted from the syringes. The extracts were analyzed by enzyme immunoassay for the presence of HIV antibodies. PCR was conducted to detect HIV and human DNA, as well as HIV RNA. RESULTS: HIV antibodies were detected in 16 (6.2%) out of 260 syringes. Human DNA or HIV-specific DNA were not detected. A second set of 80 syringes was collected to examine the presence of HIV RNA. HIV RNA was detected in three (3.8%) out of 80 syringes. CONCLUSION: This analysis demonstrates that the risk of transmitting HIV from syringes that have been used for intramuscular or subcutaneous injection may be low, but is not zero. PMID- 9863880 TI - Clinical utility of PCR on cerebrospinal fluid for the diagnosis and management of HIV neurological complications. PMID- 9863881 TI - Monitoring of pp65 antigenaemia test for cytomegalovirus after initiation of highly active antiretroviral therapy. PMID- 9863882 TI - Kaposi's sarcoma and protection from AIDS dementia complex. PMID- 9863883 TI - Pneumocystis carinii prophylaxis can be discontinued after CD4+ cell recovery over 200 x 10(6)/l. PMID- 9863884 TI - Ritonavir promotes increased growth in HIV-infected children. Paediatric AIDS Group of Switzerland. PMID- 9863885 TI - High rebound of plasma and cellular HIV load after discontinuation of triple combination therapy. PMID- 9863886 TI - Rebound of HIV-1 viral load after suppression to very low levels. PMID- 9863887 TI - Nelfinavir and nevirapine interaction? PMID- 9863888 TI - Adherence to combination antiretroviral therapy assessed by anonymous patient self-report. PMID- 9863889 TI - Signaling in human lupus T lymphocytes. PMID- 9863890 TI - Beta2GP-I in the anti phospholipid (Hughes') syndrome--from a cofactor to an autoantigen--from induction to prevention of antiphospholipid syndrome. PMID- 9863891 TI - Advances in autoantibodies in SLE. PMID- 9863892 TI - Abnormal pattern of tyrosine phosphorylation in unstimulated peripheral blood T lymphocytes from patients with systemic lupus erythematosus. AB - Previous reports have shown abnormal responses mediated via the TCR/CD3 pathway in T lymphocytes from systemic lupus erythematosus (SLE) patients. Recently, we and others have reported augmented TCR/CD3-mediated responses in lupus T cells. It is possible that the pattern of downstream biochemical signals triggered by TCR/CD3 ligation may be altered in T lymphocytes from patients with SLE, thus leading to abnormal distal cell responses. In this paper we have examined the phosphorylation of proteins on tyrosine residues in peripheral blood T lymphocytes from a group of SLE patients and controls. We found a lower frequency of constitutively tyrosine-phosphorylated 119- and 113-kDa substrates and an enhanced frequency of tyrosine-phosphorylated 66- and 25-kDa proteins in unstimulated cultures of SLE T lymphocytes, suggesting an altered pattern of tyrosine phosphorylation in T cells from patients in vivo. Additionally, the protein tyrosine phosphatase (PTP) activity of CD45 immunoprecipitates was lower in unstimulated lupus T cells and was enhanced after stimulation via the CD3 pathway in lupus but not control T lymphocytes. The present results seem to suggest abnormal regulation of in-vivo tyrosine phosphorylation in T cells from patients with SLE. PMID- 9863893 TI - SLE autoantibodies binding to native calf thymus DNA brominated in high salt. AB - Native calf thymus DNA was brominated in high salt to achieve B-->Z conformational transition. Ultraviolet and circular dichroism spectroscopic studies point towards the conformational modification of the native DNA. Specific binding of the monoclonal anti-Z-DNA antibody (Z-22) to the DNA brominated in high salt further confirmed the B-->Z conformational isomerization of native DNA. The role of Z-DNA in the etiopathogenesis of systemic lupus erythematosus has been investigated in the light of the binding of naturally occurring human anti DNA autoantibodies to the induced Z-DNA. PMID- 9863894 TI - Association of an insertion polymorphism of angiotensin-converting enzyme gene with the activity of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) shows various clinical manifestations, which are characterized by inflammation in many different organ systems. The cause of SLE is still unclear; however, the immunological abnormalities are considered to be responsible for the pathogenesis of SLE. As angiotensin I-converting enzyme (ACE) has been reported to be associated with various immunological phenomena, we investigated the correlation between insertion (I)/deletion (D) polymorphism of the ACE gene and the disease activity of SLE. Ninety-three patients with newly diagnosed SLE were enrolled in this study. ACE genotype was determined by the polymerase chain reaction (PCR). We measured serum levels of anti-double-stranded (ds) DNA antibody (Ab) and serum levels of total complements (CH50) as the parameter for lupus activity. Moreover, we evaluated the clinical disease activity by calculating SLE disease activity index (SLEDAI). Individuals with II genotype showed a significant increase in SLE activity. Patients with the ACE II genotype showed a higher serum level of anti-dsDNA Ab (14.3 IU/ml (5.475, 74.6, median (25th centile, 75th centile)) than those with the DD genotype (4.65IU/ml (4.05, 6.8)) (P<0.01). Moreover, patients with the 11 genotype also showed lower levels of serum CH50 than those with the DD genotype (P < 0.01). Patients with the II1 or DI genotype had significantly higher SLEDAI score than those with the DD genotype (P < 0.01). These results suggest that the ACE genotype could be associated with the disease activity of SLE. ACE insertion polymorphism might be used as one of predictive factors for the activity of lupus. PMID- 9863895 TI - Mucocutaneous disease in Arabs with systemic lupus erythematosus: clinical expression and relevance to autoantibodies. AB - Both criterial and non-criterial mucocutaneous manifestations of 42 Arabs with systemic lupus erythematosus are reviewed. Photosensitivity occurred in 40.5%, malar rash in 35.75% and oral ulcers in 26% of patients. Subcutaneous nodules and subcutaneous lupus erythematosus (SCLE) were not seen, and there were few cases of discoid rash (DLE), Raynaud's phenomenon, livedo reticularis and SLE-related sicca and anticardiolipin syndromes. In the clinical relevance of autoantibodies in these patients, there was a significant association between anti-Sm antibodies and oral ulcers (P= 0.033) and, interestingly, between anti-cardiolipin (aCL) antibodies and lack of photosensitivity (P = 0.014). The report also reviews previously presented data on mucocutaneous LE in Arab and non-Arab patients and emphasises the presence of intra- and inter-racial variations of SLE expression including the clinical relevance to autoantibodies. PMID- 9863896 TI - Echocardiography in systemic lupus erythematosus. AB - Two dimensional echocardiography with doppler examination was performed in 54 patients with systemic lupus erythematosus (SLE). Nine (17%) had significant cardiac involvement (four left ventricular hypertrophy, one moderate pericardial effusion, one severe aortic regurgitation, and three ventricular systolic dysfunction). We further studied diastolic function in 45 patients who did not have a major abnormality in echo. SLE was graded as active in 16 patients (SLEDAI > 5) and inactive in 29 patients. Twenty age- and sex-matched subjects acted as controls. The data were compared using one way ANOVA test. Patients with active disease had significant diastolic dysfunction compared to inactive patients and controls as indicated by increased peak A (P < 0.01) and decreased E/A ratio (P < 0.01). There was no linear correlation between disease activity and diastolic dysfunction if SLEDAI was considered as a continuous variable (r=0.29 for E/A). Anticardiolipin antibodies (both IgG and IgM) were elevated in five patients (13 studied). One of them had severe mitral regurgitation, one had trace mitral and aortic regurgitation and one had diastolic dysfunction. We conclude that asymptomatic diastolic dysfunction is present in SLE patients. PMID- 9863897 TI - Pulmonary involvement in juvenile systemic lupus erythematosus: a study on lung function in patients asymptomatic for respiratory disease. AB - Pleuro-pulmonary involvement has been well recognized in adults affected with systemic lupus erythematosus (SLE), but few studies have been carried out in children. A longitudinal study on a group of 15 children affected with juvenile SLE (JSLE), asymptomatic for lung disease, was performed, and the prevalence and the features of respiratory function alterations, over a period of 12 months, were analysed. Moreover, a possible correlation between any pulmonary function test (PFT) and disease duration, disease activity, visceral involvement and immunological pattern was evaluated. At baseline, a significant functional lung impairment was present in 40% of patients, with a significantly reduced FVC, VA and DLCO in 26% of them; in 60% of patients at 6 months and in 33% of patients at 12 months. At 6 and 12 months, our data did not show any significant modification in PFTs and the restrictive pattern, observed at baseline, remained unchanged. No correlation between altered PFTs and disease duration, activity and/or immunological findings was found. At baseline, the presence of neurological involvement was the only extra-pulmonary feature correlated to reduced FVC. PMID- 9863898 TI - Anticardiolipin antibodies in childhood rheumatic disorders. AB - Anticardiolipin antibodies (aCL) have been reported to occur in a wide variety of autoimmune and non-autoimmune disorders in adults. Our objective was to investigate the prevalence and isotype distribution of aCL and its relationship with the features of antiphospholipid syndrome (APS) in childhood rheumatic disorders. Between November 1995 and May 1996, all patients who visited our paediatric rheumatology clinic whose guardians signed a consent form participated in the study. The study population included 106 patients (36 systemic lupus erythematosus (SLE), 28 juvenile rheumatoid arthritis (JRA), 11 fibromyalgia, 7 sarcoidosis, 5 dermatomyositis, 3 rheumatic fever (RF), 3 vasculitis, 2 scleroderma, and 11 miscellaneous). aCL measurements were performed by enzyme linked immunoabsorbent assay (ELISA). All patients were carefully evaluated for symptoms and signs of APS. Eighteen of the 106 patients (17%) were tested positive for one or more of the three aCL isotypes. In SLE, aCL were found positive in 13 of 36 (37%); in JRA 2 of 28 (7%); in sarcoidosis 2 of 7; and in RF 1 of 3. aCL of IgG isotype were found positive in 16 patients (11 SLE, 2 sarcoidosis, 2 JRA, and 1 RF). This isotype was usually detected at low titers (16-24 GPL). aCL of IgM isotype were found positive in five patients (2 sarcoidosis, 2 SLE, 1 JRA), and aCL of IgA isotype were found positive in only three patients (2 SLE, 1 sarcoidosis). Clinical features of APS were rarely seen in our SLE population and were not associated with the presence of aCL. None of the patients in the other groups exhibited any clinical manifestations of APS. In conclusion, aCL were found in 37% of our childhood SLE patients as compared with only 7% in JRA. These were mostly aCL of IgG isotype of low titers and therefore were not associated with the main features of APS. Prospective studies with a larger sample size may be needed to ascertain the exact prevalence and clinical significance of aCL in childhood-onset SLE. PMID- 9863899 TI - Visual disturbance by lymphocytic hypophysitis in a non-pregnant woman with systemic lupus erythematosus. AB - The authors present the case of a patient with systemic lupus erythematosus who developed visual disturbance and amenorrhea. Though the clinical and radiological findings resembled those of pituitary adenoma, the patient was finally diagnosed as having lymphocytic hypophysitis after the operation. We briefly describe this relatively rare entity in relation to its autoimmune pathogenesis. PMID- 9863900 TI - Nervous system lupus mimics limbic encephalitis. AB - A 28-year-old woman with systemic lupus erythematosus (SLE) suffered recent onset fever, headache, encephalopathy followed by severe, repeated generalized seizures. Investigations revealed limbic encephalitis. Tests for Herpes simplex encephalitis and paraneoplastic encephalomyelitis were negative. High titers anti ribosomal-P antibodies in the cerebrospinal fluid (CSF) suggested an association with nervous system lupus. No brain biopsy was performed. Treatment was with anti seizure, anti-viral, and immunomodulating medication. PMID- 9863901 TI - Exacerbation of systemic lupus erythematosus related to cytomegalovirus infection. AB - We report two patients with systemic lupus erythematosus (SLE) in whom cytomegalovirus (CMV) infection may have played a significant role in the exacerbation or onset of symptoms. The first patient had thrombocytopenia and the second had proteinuria. CMV infection was observed in both patients when their symptoms developed. PMID- 9863902 TI - TCR AV24 gene expression in double negative T cells in systemic lupus erythematosus. PMID- 9863903 TI - Exacerbation of systemic lupus erythematosus by breast feeding. PMID- 9863904 TI - Anaplastic carcinoma of the fimbriated end of the fallopian tube as an incidental finding. AB - Carcinoma of the fallopian tube is an uncommon gynecologic tumor that is usually diagnosed in an advanced stage. The majority are tubal in origin, and rarely arise in the fimbriae. It appears that the latter may have a worse prognosis than the equivalent stage of tubal tumors that do not arise from fimbriae. We present a case of a 53-year-old white woman with FIGO stage 1 primary anaplastic carcinoma of the fimbriated end of the fallopian tube that was incidentally found in a specimen resected during a total abdominal hysterectomy with bilateral salpingo-oophorectomy. The patient underwent surgery because of findings of severe cervical dysplasia, atypia and dyskaryosis on a routine Papanicolau smear. Postoperative recovery was uneventful, and follow-up abdominal and pelvic CT scans showed no evidence of disease. However, because of the poor degree of differentiation, focal serosal infiltration and fimbrial end tube site of the carcinoma she was considered to have a high risk of recurrence. Thus, it was recommended that she undergo adjuvant chemotherapy with cyclophosphamide and carboplatin. Eighteen months after diagnosis, the patient is alive and well with no evidence of disease. PMID- 9863905 TI - Intraperitoneal contrast material combined with CT for detection of peritoneal metastases of ovarian cancer. AB - OBJECTIVE: To determine the sensitivity and specificity of CT-peritoneography (CT P) in detecting peritoneal carcinomatosis in primary or recurrent ovarian cancer. METHODS: Twenty-five patients were submitted to a standard abdominal CT (CT) as well as a computed tomography after intraperitoneal infusion of contrast material (CT-P). Twenty patients had ovarian masses clinically suspected to be malignant. In five patients with ovarian cancer who underwent prior debulking surgery, recurrent disease was suspected. RESULTS: In 21/25 patients an ovarian malignancy was histologically confirmed. During surgery peritoneal spread was found in 13 patients; only in 5 cases CT correctly suggested peritoneal metastases (sensitivity 38%). However, in 10/13 women CT-P indicated peritoneal spread, increasing the overall sensitivity from 38% to 77%. Sensitivity varied substantially according to the different abdominal areas, and was lowest in the left subphrenic space (25%). The sensitivity of CT-P was 71% and 72% in the right paracolic gutter and the pelvis, respectively. Sensitivity of CT-P was not found to be size-dependent, but was mainly related to the morphology of the lesions, with 100% sensitivity in nodular lesions, and only 21% sensitivity for flat peritoneal lesions. CT-P did not improve detection of omental metastases, and was not advantageous when ascites was present. Previous surgery reduced overall specificity from 80% to 57%. CONCLUSION: CT-P greatly improved the sensitivity of CT in the preoperative detection of peritoneal spread of ovarian malignancy. However, the technique failed to detect flat peritoneal metastases, and had a low specificity in patients with a history of prior abdominal surgery. PMID- 9863906 TI - Clear cell carcinoma of the ovary: poor prognosis compared to serous carcinoma. AB - BACKGROUND: A review of literature comparing the survival of patients with clear cell carcinoma of the ovary to patients with serous carcinoma reveals divided opinions. No studies of statistical significance have demonstrated worse survival in a cohort of patients with clear cell carcinoma matched stage for stage with patients with serous carcinoma of the ovary. The purpose of this study was to compare survival in a cohort of clear cell carcinoma patients to a cohort of serous carcinoma patients matched for stage, age, treatment, and cytoreduction. METHODS: All cases of clear cell carcinoma and serous carcinoma of the ovary operated on by the gynecology oncology service from January 1, 1981 to December 31, 1989 were evaluated for patient age, length of survival and level of primary cytoreduction, as well as FIGO stage and histology. RESULTS: Twenty-two patients with clear cell carcinoma found in the years noted were compared to a cohort of 22 patients with serous carcinoma matched for stage (I, 18.2%; II, 9.1%; III, 63.6%; IV, 9.1%), age (clear cell carcinoma 58 years, serous carcinoma 60 years (p = 0.330)), and level of primary cytoreduction (optimal in 63.6% of both clear cell carcinoma and serous carcinoma cohorts and non-optimal in 36.4% of both groups). Survival in the clear cell carcinoma cohort (16 months) was worse than in the serous carcinoma cohort (36 months) (p = 0.045). CONCLUSION: Patients with clear cell carcinoma have a significantly worse prognosis than patients with serous carcinoma when matched for age, stage, and level of primary cytoreduction. PMID- 9863907 TI - Masculinizing sclerosing stromal cell tumor in pregnancy: report of a case and review of the literature. AB - PURPOSE: To report a case of sclerosing stromal cell tumor of the ovary in pregnancy and to review the literature of this rare ovarian tumor. METHODS: The patient's office record and hospital record were reviewed. Gross and microscopic pathology were reviewed by one gynecologic pathologist. A Medline literature search and literature review were performed. RESULTS: Over eighty cases of sclerosing stromal cell tumor of the ovary have been reported in the literature since the definition of this pathologic condition in 1973. Only five cases have been reported during pregnancy and only two of these resulted in maternal virilization. This report describes severe maternal virilization with birth of a non-virilized female infant. CONCLUSION: Sclerosing stromal cell tumor of the ovary can result in severe virilization via androgen production. Surgery with removal of the involved ovary is required for diagnosis and cure. Prognosis is excellent. PMID- 9863908 TI - Correlation between immunoreactivity for transglutaminase K and for markers of proliferation and differentiation in normal breast tissue and breast carcinomas. AB - We investigated immunohistochemically localization and expression of transglutaminase K (TGK) in normal breast tissue (n = 10) and in breast carcinomas (n = 30). Transglutaminase K was compared with the staining patterns of cytokeratin 10, Ki-67, p53, estrogen and progesterone receptors in these tumors. Weak to strong membrane bound immunoreactivity to TGK was detected in 17 out of 30 breast carcinomas analyzed. TGK staining was heterogeneous with visual differences between individual tumour cells. Ninety percent of normal breast tissues revealed no immunoreactivity to TGK. Both intensity of TGK immunostaining and number of TGK-positive cells were upregulated in breast carcinomas as compared to normal breast tissue. Analyzing coexpression of TGK with cytokeratin 10, Ki-67, p53, ER and PR, no statistically significant correlation was found. Our findings indicate that: (I) TGK is upregulated in breast carcinomas as compared to normal breast tissue. (II) Upregulation of TGK in breast carcinoma is not exclusively induced by alterations of epithelial differentiation or proliferation, but by different, unknown mechanisms. (III) Upregulation of TGK in breast carcinomas may play an important role in the regulation of tumour cell invasive properties by modulating cell-matrix interactions or by facilitating the assembly of matrix and tissue remodeling. PMID- 9863910 TI - Acquired uterine arteriovenous fistulas after choriocarcinoma. A case report. PMID- 9863909 TI - Report on the antiemetic effectivity of a single 1 mg oral granisetron in moderately emetogenic chemotherapies of gynecological malignancies. AB - OBJECTIVE: On the basis of the relatively long 9-12 hour plasma-half-life of the highly selective serotonin antagonist granisetron (GRAN) and on observations that emetic episodes are detected mainly 10 hours after starting chemotherapy it was supposed that a single 1 mg oral GRAN (Kytril, SmithKline Beecham) is enough to prevent the emesis of moderately emetogenic chemotherapies. MATERIALS AND METHODS: A total of 45 gynecological cancer patients were treated in 115 chemotherapy cycles with a single 1 mg oral GRAN in a prospective, non-randomized study. Patients stayed in hospital for 24 hours after chemotherapy and emetic episodes were registered on special charts. RESULTS: A complete response (CR) of 83.4% was found in the 115 cycles. Thirty-seven of the total 45 patients with ovarian cancer had a 84.7% CR in the first 24 hours of chemotherapy. In 29 patients 50 mg/m2 cisplatin was administered with a 79.9% CR. CONCLUSIONS: 1 mg oran GRAN is a suitable antiemetic therapy in moderately emetogenic chemotherapies. PMID- 9863911 TI - Papillary squamotransitional cell carcinoma of the uterine cervix: an advanced stage disease despite superficial location: report of two cases and review of the literature. AB - Papillary carcinoma of the uterine cervix with features reminiscent of transitional cell carcinoma of urothelial origin is a poorly recognized subtype of cervical carcinoma. This tumour has a propensity for late metastasis and local recurrence in spite of the fact that histologically it could be misinterpreted as cervical intraepithelial neoplasia grade 3 (CIN3) with a papillary configuration or as a squamous papilloma. In this report we describe two examples of papillary squamous/transitional carcinoma of the cervix with literature review. The results indicate that this tumour occurs mainly in postmenopausal women, is potentially aggressive and presents at a more advanced stage inspite of the histological appearance suggesting a superficial or early invasive lesion. PMID- 9863913 TI - Primary reconstructive surgery with a sliding flap for carcinoma of the Bartholin gland. AB - Bartholin gland tumours are rare and therefore recommendations concerning management have been based on small series. Although there is a tendency today to perform less radical surgery, meaning hemivulvectomy or a wide local excision, the anatomical position of these tumours often leads to a large postoperative defect. The objective of this report is to present data on three patients but mainly to describe our surgical technique where a sliding flap obtained from the gluteal region is used for primary closure. This surgical procedure offers a functional and cosmetically acceptable vulva which is of importance considering the relative young age of patients with Bartholin gland tumours. PMID- 9863912 TI - Is there an effect of perioperative blood transfusion on the outcome of radical hysterectomy with lymphadenectomy for cervical cancer in South Africa? AB - The aim of this study was to investigate whether there is an effect of perioperative blood transfusion on the outcome of radical hysterectomy with lymphadenectomy for cervical cancer. One hundred and thirty-one patients with cervical cancer were treated by Wertheim radical hysterectomy in the period from 1984-1991. Eighty-six patients received blood transfusions during surgery or within two weeks, whereas 45 patients did not receive any blood transfusion. Transfused and non-transfused patients did not differ with respect to mean age, race, weight, FIGO-stage, cell-type, grade, size, depth of invasion and nodal involvement. Transfused patients had more blood loss, longer surgical time and lower haemoglobin levels. Using log rank analysis, the calculated five-year survival was 81% for the transfused group and 84% for the non-transfused group, a non-significant difference. The five-year disease-free survival rate was 87% for the transfused group and 88% for the non-transfused group. This study suggests that perioperative blood transfusion does not adversely influence survival after the Wertheim operation for cervical cancer. PMID- 9863914 TI - HPV and intraepithelial neoplasia recurrent lesions of the lower genital tract: assessment of the immune system. AB - OBJECTIVE: To evaluate the immune state in patients with genital relapse HPV and intraepithelial lesions of the lower genital tract. METHOD: Forty-three patients were selected. Twenty-one were affected by recurrent HPV infection either alone or combined with intraepithelial neoplasia treated by laser surgery, and 22 had been previously-treated and clinically cured without recurrence during a follow up from 18 to 24 months. The diagnostic protocol included colposcopy with eso- and endocervical cytology histologically confirmed by directed biopsy. Afterwards patients underwent a systemic immunogenic evaluation. RESULTS: NK cell reduction was strictly related to HPV infection associated with intraepithelial lesions; B lymphocyte reduction was percentually greater in patients affected by HPV alone; activation of R-IL2 increased in a percentage overlapping in the two groups indicating patient reaction to the virus. CONCLUSION: Our study supports the theory that immune response directed against viral antigens is one of the most important effectors in the control of HPV infections and that HPV is the cause of a systemic rather than local lesion. PMID- 9863915 TI - Leiomyosarcomas of the female genital tract: a clinical and histopathological study. AB - INTRODUCTION: Leiomyosarcomas are malignant tumours showing smooth muscle differentiation. They represent approximately 25% of all uterine sarcomas and slightly over 1% of all uterine malignancies. The purpose of the present retrospective review is to relate clinical and pathological findings of leiomyosarcomas of the female genital tract to prognosis. MATERIAL AND METHODS: During 1972-1992 eleven patients had diagnosed uterine leiomyosarcomas treated at the Department of Gynecology of the University of Saarland. The hospital records of all patients were reviewed and complete primary treatment had been performed at this center. RESULTS: The mean age was 46.92 years (SD: +/-13.85). Atypical uterine bleeding and pelvic discomfort were the most common presenting complaints (72.7%). The mean follow-up time was 59.60 months (20-96 months). Overall 2-year survival was 70% and overall 5-year survival 30%. The overall survival of patients in FIGO-stage I was 57.14%, in FIGO-stage II 100%, in FIGO-stage III 0% and in FIGO-stage IV 0%. CONCLUSION: The primary therapy should consist of an operation as radical as possible. Treatment with organ preserving seems to be reasonable if the patient desires children. Also, chemotherapy might provide a hopeful sign in the improvement of survival rates. PMID- 9863916 TI - Tamoxifen-induced endometrial polyp. A case report and review of the literature. AB - A case of an endometrial polyp which developed in a 74-year-old woman treated with tamoxifen for 15 years after breast cancer surgery was the stimulus for this brief and concise review of the endometrial changes caused by anti-estrogen treatment in post-menopausal women with breast cancer. Tamoxifen therapy has been associated with the development of endometrial polyps, hyperplasia and adenocarcinoma possibly mediated through its agonistic estrogenic properties. Hysteroscopy follow-up should be performed in this group of patients and hysteroscopy should be done before the beginning of therapy and repeated once a year during the treatment. PMID- 9863917 TI - Granulocyte macrophage-colony stimulating factor for treatment of chemotherapy extravasation. PMID- 9863918 TI - A 30s PAS stain for frozen section analysis of surgical margins of vulvectomy in Paget's disease. AB - Application of a modified periodic-acid Schiff's procedure (PAS) technique in frozen section analysis of surgical margins of vulvectomy specimen in Paget's disease is described. The reaction obtained using a microwave procedure takes only 30 seconds. This method was found to be useful in identifying Paget cells on the margins thought to be free of disease by conventional histological evaluation. PMID- 9863919 TI - Significance of tissue polypeptide specific antigen (TPS) in diagnosis and monitoring of treatment in ovarian cancer. AB - Tissue polypeptide specific antigen (TPS) finds increasingly broad application in diagnosis and monitoring of treatment in ovarian cancer. Its sensitivity increases in parallel to clinical advancement of the tumor and to the grade of cellular differentiation. TPS may represent valuable supplementation of conventional markers in diagnosis of mucous carcinoma. Improved sensitivity and specificity of the technique for detection of ovarian cancer occur when two or more additional tumour markers are used in parallel. TPS has been found to represent a dependable index of surgical completeness. Determination of TPS in the sera of patients with ovarian cancer in the course of chemical treatment provides important information on the course of the neoplastic process and defines the response of the host to the applied treatment. PMID- 9863920 TI - Breast cancer during pregnancy and lactation. AB - Three cases of breast cancer during pregnancy and lactation which were referred to the Obstetrics and Gynecology Department of loannina University Hospital during the period 1990-1997 are presented. Diagnosis and management of these cases are discussed and a strength protocol is suggested to identify new cases of breast cancer during pregnancy and lactation. PMID- 9863921 TI - Endometrium adenocarcinoma with choriocarcinomatous differentiation: a case report. PMID- 9863922 TI - The frequency of Chlamydia trachomatis infection in women with intraepithelial neoplasia of the uterine cervix. AB - OBJECTIVE: To evaluate the frequency of Chlamydia trachomatis (CT) infection in patients with cervical intraepithelial neoplasia (CIN) referred for conisation. METHODS: 423 patients with histologically proven CIN were included in this study. Before conisation, cytological material for direct immunofluorescence analysis was obtained. The incidence of CT in cytologic smears of all three CIN groups was calculated. RESULTS: Among all patients with CIN, 27 (6.4%) were CT positive and 396 (93.6%) CT negative. There were 2 (8.3%) CT positive patients with CIN 1, 10 (9.3%) with CIN 2 and 15 (5.2%) with CIN 3. The difference between CT infection incidence in patients with CIN 1 and CIN 2 was not significant; neither was it significant between patients with CIN 1 and CIN 3 nor between those with CIN 2 and CIN 3. CONCLUSIONS: Chlamydia trachomatis infection in patients with CIN is not very common. The impact of CT infection does not seem to interfere with development or even promotion of CIN. PMID- 9863923 TI - Screening of persistent trophoblastic disease with various serum markers. AB - Eighty percent of the patients with molar pregnancy go into spontaneous remission and do not require any therapy. Serial hCG determinations can identify the 20% who will develop malignant sequelae. It does not seem appropriate to treat all patients. This study was designed to assess several serum markers, including free beta-hCG, total beta-hCG, and CA-125 in order to identify persistent trophoblastic disease. The study was performed at Doctor Zekai Tahir Burak Women's Hospital, Department of Oncology. Forty-seven patients with complete hydatidiform mole were included in the study. In the spontaneous remission group (Group I), total betahCG, CA-125 and free betahCG values were 27988.7+/-18491.6 mlU/ml, 51.7+/-74.7 U/ml and 42.35+/-28.4 mlU/ml, respectively. Patients in whom persistent trophoblastic disease had developed (Group II) the mean serum CA-125 and mean total betahCG values were lower than in group I, whereas the mean free betahCG value was higher but not significant. The mean value of free betahCG per total betahCG was found to be significantly higher in group 2. The free betahCG per total betahCG ratio seems to be a sensitive predictor of persistency of trophoblastic disease. Further prospective studies with a larger series of patients may warrant the exact predictive value of free betahCG per total hCG ratios. PMID- 9863924 TI - Unexpected increase of the CA 19-9 tumour marker in patients with endometriosis. AB - The purpose of this study was to investigate whether the ovarian function of women with endometriosis could be identified by serologic concentration of the oncofetal antigen CA 19-9 before, during and after treatment for 6 months with danazol. A total of 15 women with endometriosis, 20-40 years old, were studied. The serum CA 19-9 antigen was measured by immunoradiometric assay. The measurement of CA 19-9 was repeated during the last 15 days of a 6-month therapy period with danazol and after 3 months from the end of the therapy in only seven women. It was found that: 1) Eight out of 15 women (53.3%) showed higher CA 19-9 values than the upper normal limit. 2) Danazol caused a significant decrease in the antigen values which remained stable after the cessation of therapy. This data suggests that endometriosis should be classified among other diseases which cause an increase in the serum levels of CA 19-9 and that ovarian function is related to antigen levels. PMID- 9863925 TI - p53 expression in ovarian carcinoma with regard to second-look findings. AB - Forty-eight (48) cases of ovarian carcinoma subjected to primary surgery and platinum based chemotherapy followed by second-look laparotomy (SLL) were analyzed with respect to p53 status. Overall aberrant p53 accumulation was observed in 20 (41.6%) out of 48 patients. While one out of 5 patients (20.0%) with stage I-II disease showed positive p53 staining, 19 out of 43 patients (44.1%) with stage III disease were found to be positive for p53. According to the histologic types, 41.9% of serous (13/31), 33.3% of endometrioid (2/6), 50.0% of transitional (2/4), 40.0% of mixed (2/5) and 50,0% of mucinous types of tumors showed abnormal expression of p53. p53 positivity was found in 12 (54.5%) out of 22 SLL(+) patients while only 8 (30.7%) were p53 positive out of 26 with SLL(-) patients. Cases with abnormal p53 accumulation tended to be SLL positive, however, the difference was not significant. PMID- 9863926 TI - The value of tissue polypeptide specific antigen TPS determination in serum of women with breast cancer comparison to mucin-like associated antigen MCA and CA 15-3 antigen. AB - The aim of the study was TPS diagnostic value determination in the serum of women with breast cancer as compared with MCA and CA 15-3. The relationship between the serum concentration of these antigens and patient age, clinical stage, histological grade, presence of metastases to lymph nodes and histological type of neoplasm was evaluated. Studies were conducted on the sera of 139 women before surgical procedure aged 28-81, treated in the Clinic of Oncology, at Karol Marcinkowski University of Medical Sciences in Poznan. The TPS concentration was determined using the "BEKI Diagnostics" immunoenzymatic method, MCA - by the "Roche" test and CA 15-3 concentration was determined by the "Abbott" immunofluorescent test. The study showed significantly higher levels of TPS, CA 15-3 and MCA in women with cancer, compared with values in healthy women and women with mastopathy. The highest median of concentration and frequency of occurrence was obtained for TPS. A correlation between enhancement of TPS and CA 15-3 concentration with clinical stage was observed. A similar connection was noted in women with metastases to the lymph nodes. Serum MCA concentration results did not demonstrate the above effects. The study suggests, that in estimating the clinical condition of women with breast cancer, the simultaneous determination of TPS with CA 15-3 seems to be a more useful prognostic factor than TPS or CA 15-3 with MCA. PMID- 9863927 TI - Skinning vulvectomy for the treatment of vulvar intraepithelial neoplasia 2-3: a study of 21 cases. AB - Twenty-one cases of patients with vulvar intraepithelial neoplasia (VIN) 2-3 were reviewed. The mean age at diagnosis was 45.4 years. All of the patients presented with vulvar pruritus. Five of the patients had hypertension, two had coronary heart disease and two had diabetes mellitus as complicating medical illnesses. None of the patients had history or evidence of vaginal intraepithelial neoplasia (VAIN) or cervical intraepithelial neoplasia (CIN), and only one patient had invasive cervical cancer at diagnosis. Provided the histology confirmed VIN, the patients were subjected to a skinning vulvectomy procedure. Of the patients, 15 (71.4%) had VIN 2, and the remaining 6 (28.6%) had VIN 3 at preoperative evaluation. Histologic analysis of skinning vulvectomy specimens revealed no evidence of neoplasia in three patients (14.2%). Multifocality was observed in only three patients (14.2%). The areas involved were the perineum in four patients, labia in 15 and clitoris in two patients. Associated vulvar pathologies were condyloma acuminata in one, squamous vulvar hyperplasia in three and lichen sclerosus with squamous hyperplasia in one patient. The complications of the procedure included febrile morbidity in three patients and minor wound break-down in one patient. None of the patients in this series experienced recurrence. Skinning vulvectomy seems to have a high success rate in treatment of VIN 2-3 with minimal postoperative complications and satisfactory cosmetic results. However, observation of only three patients with multifocal lesions as well as no patient with invasive cancer adds credence to an ablative procedure after appropriate evaluation under colposcopy. PMID- 9863928 TI - Donal Munro Lecture: Functional and neurologic recovery following acute SCI. AB - Patient management in the current health care environment requires the provider to make reliable and valid clinical decisions regarding patient care, decisions that are cost effective and will lead to optimal functional improvement. Neurologic examination, according to the Standards for Neurological and Functional Classification of Spinal Injury developed by the American Spinal Injury Association, provides every clinician with simple clinical tools that are highly predictive of functional recovery following a spinal cord injury. The ability to predict motor recovery provides a rational basis to help support the provider's recommendation for the patient's goals and care planning, as well as differentiates care that is essential for improving functional outcomes from care that is not. PMID- 9863929 TI - 4-Aminopyridine alters gait characteristics and enhances locomotion in spinal cord injured humans. AB - Recovery of useful motor function in humans with spinal cord injury (SCI) is a primary and elusive goal. In this preliminary study, we describe efforts to delineate the pharmacological effects of 4-aminopyridine (4-AP) on gait parameters in spinal cord injured humans who have retained some capacity to ambulate bipedally. A sequential entry, open label study was made of the effects of a single oral administration of an immediate-release formulation of 4-AP on the time-course profile of changes in component parameters of bipedal gait in ambulatory volunteers with chronic SCI. Nine healthy, rehabilitated, community adapted male volunteers (six tetraparetic, three paraparetic), who sustained their injuries more than one year prior to entry into the study, ingested a single 10-mg dose of 4-aminopyridine after an overnight fast. Gait analysis parameters included velocity (meters/min), cadence (steps/min), stride length (meters), gait cycle (seconds), and double limb support (percent of gait cycle). They were measured for 24 hours using a sampling-rich strategy (nine duplicate measurements over 24 hrs). Repeated measures (randomized block) analysis of variance (ANOVA) and paired t-tests were used to test for the significance of differences between means and variances. The apparent pharmacological effect of 4 AP is associated with statistically significant changes in one or more of the component elements used to assess the characteristics and efficiency of bipedal gait. These changes in gait analysis parameters correspond temporally with the improvements in pulmonary function and heart rate variability previously described by us. 4-AP appears to enhance gait in a subset of humans with SCI. In this preliminary study we report, for the first time, an apparent effect of 4-AP on gait in spinal cord injured humans and suggest that the pharmacological effects of 4-AP may have clinically significant application in the restoration of useful motor function. PMID- 9863930 TI - Treatment of acute deep vein thrombosis in spinal cord injured patients with enoxaparin: a cost analysis. AB - Enoxaparin, a low molecular weight heparin, has been demonstrated to be effective in the prophylaxis and treatment of deep vein thrombosis (DVT) in the general population. This study presents an analysis of the costs of subcutaneous (SQ) enoxaparin compared with intravenous (IV) heparin. Data were gathered on six spinal cord injured (SCI) patients in an acute freestanding rehabilitation center who were initially treated with SQ enoxaparin (n=3) or IV heparin (n=3) for a proximal DVT. No patients in either group developed further clinical complications. Comparison of the total costs of each treatment was performed, including the direct cost of the drug, as well as some of the costs of administration. Although the cost of enoxaparin per unit dose is higher, the total costs of enoxaparin are slightly lower, because its labor and administration costs are less. Subcutaneous enoxaparin is a safe, cost-effective, and less labor-intensive treatment, and can be of substantial benefit in the treatment of DVT in SCI patients in the rehabilitation setting. PMID- 9863931 TI - Optimization of sacral ventral root stimulation following SCI: two case reports with six-month follow-up. AB - Sacral ventral root stimulation in conjunction with sacral dorsal rhizotomy has been effective in promoting voiding in individuals with upper-motor-neuron spinal cord injury. We report on two patients who had variable voiding responses to stimulation during the first six months after electrode implantation. We used videourodynamic records and daily voiding records to characterize their voiding difficulties. Different methods were used to improve voiding, including seating adjustments and changes in stimulation parameters. The first patient was unable to empty his bladder on a regular basis with stimulation using 24 pulses per sec stimulating frequency for the first two months after implantation. Voiding was substantially improved by using 35 pulses per sec. At the end of six months, he is regularly emptying his bladder with stimulation and is on an every-second-day bowel program. However, his bowel program has been irregular. The second patient had very good voiding when stimulation was applied in bed, but he had poor voiding with high residual volumes when sitting in his wheelchair. Voiding was improved when he used a wheelchair cushion that was cut out in the back or lifted his buttocks off the chair. These procedures appeared to reduce perineal pressures. This patient has bowel care on alternate days and his bowel care time has been reduced following implantation of the device. Neither of the patients experienced an erection with the device. Both patients feel positive about their implant experience. PMID- 9863932 TI - Self-reported use of an implanted FES hand system by adolescents with tetraplegia. AB - The Freehand System, an eight-channel functional electrical stimulation (FES) system, was implanted in five adolescents with C-5 or C-6 tetraplegia to provide stimulated lateral pinch and palmar grasp. Following completion of inpatient training on how to use the Freehand System for predefined and self-selected activities of daily living (ADL), the adolescents were discharged to use their Freehand systems at home and school. A telephone survey was administered on a weekly basis to obtain information on the type of ADL performed with the Freehand System, reasons for not using the Freehand System, and perceived barriers and motivators to FES use. Twenty surveys were obtained on each adolescent, resulting in a total of 100 surveys. The most common type of activity performed with the Freehand System was self-care, which included tasks such as eating, grooming, and brushing teeth. The Freehand System was also used for productivity activities defined as writing, socialization, and manipulation of school and household objects. Motivators to Freehand System use included perceived need and importance to perform an ADL in an independent fashion, physical ease of using the Freehand System, and availability of social supports that facilitated Freehand System use. While there were several reported barriers to Freehand System use, incompatibility with multiple transfers to and from the wheelchair and lack of physical assistance during morning care to don the system were perceived as two of the more common reasons for nonuse. PMID- 9863933 TI - Botulinum toxin injections for voiding dysfunction following SCI. AB - Botulinum toxin (BT) injections have been used successfully to treat spastic muscle conditions, including detrusor-sphincter dyssynergia (DSD) seen in spinal cord injury (SCI) patients. In our urology clinic, we used BT to treat three SCI patients who had voiding dysfunction, using a transperineal needle with electromyographic (EMG) monitoring. Two of the patients reported excellent results following the treatment. One patient, with whom the staff had difficulty doing intermittent catheterization (IC), improved significantly. The other patient had improved voiding with an external catheter and minimal urinary residual. The third patient had no improvement of leg spasms with his voiding dysfunction and required a sphincterotomy. Although patients may need repeat injections, BT is minimally invasive and easy to administer with no side effects. Overall, BT injection is an excellent method of managing voiding in SCI patients, especially those on continuous external catheters and with IC management who refuse or are not good candidates for surgery. PMID- 9863934 TI - Bladder cancer risk in patients with spinal cord injury. AB - Bladder cancer (BC) is the fourth most common cancer in men. It is associated with several risk factors (RF), of which only a few have been evaluated in previous studies. Models incorporating only one or a severely restricted number of RFs did not predict BC well. We employed 19 a priori RFs and 12 interactions in a multivariate logistic regression analysis for the prediction of BC in a sample of subjects with spinal cord injury (SCI), of whom 149 were outpatients (7 with BC) and 4 were inpatients with BC. We also replicated dichotomous predictions for 10 of the 19 RFs that have been most frequently associated with a higher BC risk in the literature. The overall test for 31 predictors was significant (p = 0.0038). A dichotomized predictor correctly identified 9 of 11 BC cases and all 142 but one of the cases without BC. A more parsimonious subset of 21 predictors satisfied a Scheffe-type multiple comparison criterion. Although duration of SCI satisfied a Bonferroni criterion for statistical significance, it did not satisfy a Scheffe criterion. In the replication studies, only dichotomized duration of indwelling catheterization for at least 10 years significantly replicated the previous findings. Results of this study suggest that using multiple risk factors and interactions in a comprehensive statistical model may provide useful screening of patients with SCI for BC risk. Since early identification of BC substantially improves prognosis, such a model may identify patients at highest risk who are most likely to benefit from bladder biopsy. PMID- 9863935 TI - Sexual satisfaction and sexual drive in spinal cord injured women. AB - Levels of sexual satisfaction and sexual drive in women with spinal cord injuries were examined. Eighty-four spinal cord injured (SCI) women and thirty-seven able bodied (AB) control subjects completed the Derogatis Sexual Functioning Inventory which measured current level of sexual functioning in 10 areas: information, experience, drive, attitude, psychological symptoms, affect, gender role definition, fantasy, body image, and sexual satisfaction. When compared with AB women, SCI women had significantly lower levels of sexual satisfaction and sexual drive and significantly higher levels of psychological symptoms and negative affect. Among SCI women, sexual satisfaction decreased significantly with age. Among AB women, sexual satisfaction increased significantly with age. Married SCI women were no longer less sexually satisfied than AB women. Results support conclusions from previous self-report studies in which significant decreases in sexual satisfaction and drive were reported for SCI women. Implications for continued research on the psychological and physiological aspects of sexuality and sexual functioning in SCI women are discussed. PMID- 9863936 TI - Malone antegrade continence enemas for autonomic dysreflexia secondary to neurogenic bowel. AB - A 42-year-old man with T-4 ASIA B thoracic paraplegia presented with chronic constipation, fecal incontinence, and bowel-related autonomic dysreflexia (AD) refractory to conservative bowel program. His usual toileting time lasted one to five hours. Antegrade continence enemas (ACE) were performed. The ACE technique, which requires creating a continent catheterizable appendicocecostomy, is described. Postoperatively, a daily enema was given through the stoma. The patient's toileting time was reduced to 20 minutes and his AD resolved. The role of the ACE is discussed in adult spinal cord injury (SCI). PMID- 9863937 TI - Clinical practice guidelines: Neurogenic bowel management in adults with spinal cord injury. Spinal Cord Medicine Consortium. PMID- 9863938 TI - In situ degradation: a new concept for system suitability tests in monographs of the European Pharmacopoeia. AB - Monographs of the European Pharmacopoeia describe in the LC-test for related substances usually a system suitability test in order to ensure the adequate separation of impurities. Since the reference substances required are often not available a recent approach to avoid this problem is the generation of the required impurity by 'in situ degradation' of the active principle. This paper describes some typical applications of this technique as well as recent examples, such as the controlled degradation of cefalotin sodium, imipenem and spiramycin. PMID- 9863939 TI - Separation of steroidal drugs on porous graphitized carbon column. AB - The retention time of 11 steroidal drugs was determined on a porous graphitized carbon (PGC) column using water tetrahydrofuran mixtures as eluents. Linear correlations were calculated between the logarithm of the capacity factor (k') and the tetrahydrofuran concentration in the eluent. To find the molecular parameters significantly influencing the retention the intercept and slope values of the above relationship were correlated with the physicochemical characteristics of steroidal drugs using principal component analysis (PCA). Both the slope and intercept values show high differences proving that steroidal drugs can be successfully separated on a PGC column. Five principal components explained > 90% of the total variance. Calculations indicated that both electrostatic interactions and sterical conditions can influence the retention of steroidal drugs on PGC column. PMID- 9863940 TI - Multivariate calibration: applications to pharmaceutical analysis. AB - The principles of multivariate calibration (MC) are presented, with reference to the main objectives of this chellometrics technique: the reduction of the variance in the prediction of a response variable (generally, a chemical quantity) and the possibility of the determination of the response in complex matrices with no or limited sample preparation, as in the case of the determination of a drug in a medicament. In both cases MC uses the whole information in a spectrum (a series of predictors). The possibility of the improvement of the MC performances, eliminating some useless, noisy, predictors is shown. Variable selection has been performed using two original techniques: a stepwise elimination procedure, based on the normalised coefficients of the regression equation relating the response to the predictors and a technique based on iterative repetitions of the regression technique (partial least squares regression, PLS), each time by weighting the predictors by their normalised regression coefficient computed in the previous cycle. These strategies are illustrated by means of different data sets, a synthetic example and a real example where MC, applied to near infrared spectroscopy, is used in the analysis of a drug. In this case also the application of an original MC technique is shown, where a joint regression model is obtained for two different instruments. PMID- 9863941 TI - Application of standardisation methods to correct the spectral differences induced by a fibre optic probe used for the near-infrared analysis of pharmaceutical tablets. AB - Near infrared spectroscopy has become very popular in the pharmaceutical industry because of many important practical advantages. With the help of powerful chemometric techniques, multivariate calibration models are developed, relating near-infrared spectra to the values to be modelled. However, because of small instrumental differences between near-infrared spectrometers, a calibration model can only be used with spectra collected on the same instrument, which represents a serious limitation for the use of near-infrared spectroscopy in the pharmaceutical industry. To deal with this important problem, a certain number of different standardisation approaches were proposed in the literature. In this article, an application of instrument standardisation methods is presented, where two different data measurement modules (internal measurement cell and external fibre optic module) of a near-infrared spectrometer must be standardised for the quantitative determination of an active compound in pharmaceutical tablets. PMID- 9863942 TI - Ruggedness testing of chromatographic methods: selection of factors and levels. AB - The first step in a ruggedness test is the selection of factors to be examined and their levels. In this paper, both topics are discussed, thereby completing a strategy described earlier. It is demonstrated, by means of some examples, that depending on the formulation (definition) of a factor, information that is physically more or less meaningful is extracted from the experimental design results. Among others, the inclusion of the compounds of a buffer and of the components of a mixture in a screening design were examined. A general guideline to select the levels of the factors in a ruggedness test was proposed. Some special cases, i.e. asymmetric intervals around the nominal level, were also discussed. PMID- 9863943 TI - Experimental design for a pharmaceutical formulation: optimisation and robustness. AB - In pharmaceutical industries, the formulator is usually faced with the optimisation of the excipient mixture composition aimed to prepare a product with the required characteristics. Experimental research methodology represents an efficient approach for solving such optimisation problems. Planning mixture experiments using specific designs allows to analyse the blending properties of each mixture component and estimate an empirical model approximating the response of interest as a function of excipient proportions. In this study the evolution of theophylline solubility in a four-component system with constraints was analysed using two mixture design approaches: a classical mixture component proportion approach and a mathematically independent variable approach. An optimal region characterised by high solubility values was found and further explored in order to verify the insensitivity of theophylline solubility to slight variations of the excipient mixture composition. PMID- 9863944 TI - Set-up and validation of an adsorptive stripping voltammetric method for kynurenic acid determination in human urine. AB - Validation of an adsorptive stripping voltammetric method for kynurenic acid determination in urine, was presented. The selection of appropriate validation parameters, the design consideration for evaluation and the problem of endogenous metabolites were discussed. The considered fundamental criteria for assessing the reliability and overall performance of the method in the urine matrix were selectivity, linearity and range, limit of quantitation, accuracy, precision and analyte stability. The intermediate precision was also evaluated by means of a full factorial design. An HPLC method with fluorimetric detection was used as a reference method to assess the accuracy. The analysis in urine required a pH control as pointed out by robustness testing and the found kynurenic acid concentration in daily urine ranged from 5 to 40 microM. PMID- 9863945 TI - The voltammetric study of 2-mercapto-5-phenylammino-1,3,4-thiadiazole using carbon paste electrodes. AB - A rapid and accurate voltammetric method for the quantitative determination of 2 mercapto-5-phenylammino-1,3,4-thiadiazole (MPATD) with carbon paste electrodes (CPE) has been developed. The study was made by cyclic voltammetry between -0.4 and +0.6 V with 50 mV s(-1) sweep rate in aqueous solution. After successive oxidation/reduction cycles we found a total oxidation of MPATD at +0.45 V. As the compound is oxidated, the reduction current peak increases at +0.13 V, indicating an irreversible process. Following only the oxidation process in the -0.1 to +0.6 V range, we investigated the optimum scan rates at different current densities and pH values (realised with buffers, pH between 1.0 and 10.0) with CPE versus Ag/AgCl reference electrode using linear sweep voltammetry. We found a good linear relation between the current peak height and concentration in a 2.5 x 10( 9)-1.25 x 10(-7) mol ml(-1). This method allows the quantitative detection of the MPATD as it or from dosage forms and biological media. PMID- 9863946 TI - Conductometric method for the quantitative analysis of Pb(II) and Cd(II) with 2 mercapto-5-R-amino-1, 3, 4-thiadiazole derivatives. AB - The reactions of the cations with 2-mercapto-5-R-amino-1,3,4-thiadiazole derivatives were studied conductometrically with the purpose of establishing a new conductometric method for the quantitative analysis of Pb(II) and Cd(II). Aqueous solutions of Pb(NO3)2 and Cd(CH3COO)2 were titrated with hidroalcoholic solutions of 2-mercapto-5-amino-1,3,4-thiadiazole (MATD), 2-mercapto-5-allylamino 1,3,4-thiadiazole (MA1ATD) and 2-mercapto-5-acetylamino-1,3,4-thiadiazole (MAcATD) and 2-mercapto-5-phenilamino-1,3,4-thiadiazole (MFATD) in different concentrations. The reactions takes place at pH 6.5 (realised with acetate buffer). A linear classical titration curves was obtained. In solutions more concentrated than 10(-2) M just one equivalence point can be noticed, corresponding to 1:2 Me:R stoechiometries. For concentration less than 10(-2) M two equivalence point were observed at 1:1 and 2:1 ratio of Me:R, indicating the step formation of the complex. Accurate conductometric determinations can be made using the second break points of the titration curves as equivalence points. The amounts of Cd(II) and Pb(II) taken and recovered are good, with an error less than 1%. PMID- 9863947 TI - Cefuroxime selective electrodes for batch and FIA determinations in pharmaceutical preparations. AB - Different cefuroxime selective electrodes, without internal reference solution and comprising PVC membranes, were constructed and evaluated. Membranes were prepared with cefuroxime tetraoctylammonium (A) or cefuroxime bis(triphenylphosphoranylidene)ammonium (B) as ion-exchanger, 2-nitrophenyl octyl ether (X) or bis(2-ethylhexyl)sebacate (Y) as plasticizing mediator solvent and 4 tert-otcylphenol (TOP) as additive. From the comparative evaluation of the described electrodes, membranes comprising 2-nitrophenyl octyl ether, cefuroxime tetraoctylammonium and 4-tert-otcylphenol presented better working characteristics. For these electrodes (type XA-TOP), with a lifetime > 5 months, a lower limit of linear range of 2.8 x 10(-4) M, a practical detection limit of 1.3 x 10(-4) M, a reproducibility of approximately +/-0.6 mV day(-1) and a slope of -50.4 mV decade(-1), under H3PO4/NaH2PO4 solutions (pH 3.5; I =0.1 M), were found. The presence of the additive on the membranes was of crucial importance for the electrodes good characteristics. Interference from sulphate, chloride, nitrate, iodide, cefaclor, cefadroxil, cefazolin and cephradine, on the electrodes behaviour was evaluated. Only a slight interference from nitrate and iodide was recorded, being type XA-TOP electrodes the most selective units. Electrodes with a tubular configuration prepared with type XA-TOP membranes, aiming flow injection analysis, were also constructed. When these tubular potentiometric detectors were evaluated in a double-channel flow injection manifold, with 3.5 pH and 0.1 M ionic strength conditions, significantly better working characteristics than those of the corresponding conventional electrodes, namely higher slopes (-54.6 mV decade(-1)) and better reproducibilities (+/-0.2 mV day(-1)), were found. Both conventional and tubular type XA-TOP electrodes were used for injections analyses by batch and FIA, respectively, presenting low consumption of samples and reagents. Relative error deviations to the reference procedures <3.0% were found. PMID- 9863948 TI - Hydrogen peroxide determination in pharmaceutical formulations and cosmetics using a new catalase biosensor. AB - The possibility of evaluating the content of hydrogen peroxide in several authentic matrices, such as cosmetic and pharmaceutical formulations, was studied. A new catalase biosensor fabricated using an amperometric gas-diffusion oxygen sensor as electrochemical transducer and the catalase enzyme immobilized in kappa-carrageenan gel and capable of operating in both aqueous and non aqueous solvents was developed and tested for this purpose. Creams, emulsions and disinfectant solutions were analysed. To this end, a preliminary check was needed to establish the best conditions to analyse these matrices; the choice of solvent was one of the most important points studied. The solvents considered included dioxane, water-dioxane mixtures, water saturated chloroform and aqueous solutions. The different solubility properties of the matrices analysed were taken into account. PMID- 9863949 TI - Lidocaine and benzalkonium analysis and titration in drugs using new ISFET devices. AB - Two new ISFETs recently developed by us have now been applied to some pharmaceutical determinations in real matrices; the first device, responsive to cationic surfactants, was employed in the determination of benzalkonium chloride contained in two different disinfectant solutions and in three types of commercial collyrium; the second device, responsive to cocaine hydrochloride, showed an appreciable response also to lidocaine hydrochloride and was used in the determination of lidocaine hydrochloride contained in some injectable antibiotics. The repeatability and accuracy of measurements performed in the analysis of these pharmaceutical matrices using new solid state sensors were evaluated. A further aspect of the research involved the use of two sensors to record complete titration curves for the determination of benzalkonium chloride, cocaine hydrochloride and lidocaine hydrochloride, respectively. Applications to real matrices were also performed by analysing by titration pharmaceutical formulations containing benzalkonium chloride, or lidocaine hydrochloride and an illicit powder containing cocaine hydrochloride and sugars. PMID- 9863950 TI - Chemical modification of human albumin at cys34 by ethacrynic acid: structural characterisation and binding properties. AB - Derivatization of the free cys3,4 in human albumin, which is reported to occur under physiological conditions, has been performed in vitro by reaction of the protein with ethacrynic acid. This modification has been investigated by mass spectrometry and circular dichroism. Ethacrynic acid has been proven to bind human albumin either covalently and non-covalently. This post-translational modification does not determine significant changes in the secondary structure of the protein, as shown by the comparable circular dichroism spectra of the native and the modified proteins. Furthermore, the binding properties of the human albumin samples have been investigated by circular dichroism and equilibrium dialysis. The affinity to the higher affinity binding sites does not change either for drugs binding to site I, like phenylbutazone, or to site II, like diazepam, while a small but significant increase has been observed for bilirubin, known to bind to site III. Nevertheless significant decreases of the affinity at the lower affinity binding sites of the modified protein were observed for both drugs binding to site I or to site II. PMID- 9863951 TI - The potentiometric and spectrophotometric determination of dissociation constants for same 2-mercapto-5-R-amino-1,3,4-thiadiazole derivatives. AB - In order to establish the dissociation constants of organic compounds, the spectrophotometric and potentiometric methods are the most precise and useful ones. Comparing the results, we used both methods for three derivatives of 2 mercapto-5-R-amino-1,3,4-thiadiazole. For the pKa determination by the spectrophotometric method we measured the ratio between the concentration of dissociated and undissociated forms. The ratio was calculated from spectral data. The potentiometric method for pKa consists of measuring the pH values within a potentiometric titration with 0.1 M NaOH. The results proved a very low acid character of the derivatives. The pKa values were influenced by the properties of the amino group substituents, and the results confirm the theoretical considerations. The study confirms the thion-thiolic tautometry of the 2-mercapto 5-R-amino-1,3,4-thiadiazole derivates and their property of being ligands for the coordination of the cations of some representative and transitional metals, with application in the preconcentration, in the detection and the quantitative determination of polluting and toxic cations in environmental analysis. PMID- 9863952 TI - Determination of fleroxacin in human serum and in dosage forms by derivative UV spectrophotometry. PMID- 9863953 TI - Compatibility study between ibuproxam and pharmaceutical excipients using differential scanning calorimetry, hot-stage microscopy and scanning electron microscopy. AB - Differential scanning calorimetry (DSC) was used as a screening technique for assessing the compatibility of ibuproxam with some currently employed pharmaceutical excipients. The influence of processing effects (simple blending, cogrinding or kneading) on drug stability was also evaluated. On the basis of DSC results, ibuproxam was found to be compatible with corn starch, avicel and sodium carboxymethylcellulose. Some drug-excipient interaction was observed with polyethyleneglycol 4000, palmitic acid, stearic acid, Ca and Mg stearate. Actual solid-phase interactions of the drug with polyvinylpolypyrrolidone and polyvinylpirrolidone K30 were induced by mechanical treatments. Hot-stage microscopy (HSM) and scanning electron microscopy (SEM) were of help in interpreting the DSC results and excluding in all cases relevant pharmaceutical incompatibilities. PMID- 9863954 TI - The equilibrium constant of beta-cyclodextrin-phenolphtalein complex; influence of temperature and tetrahydrofuran addition. AB - The temperature influence on creation of a supramolecular complex in which beta cyclodextrin (beta-CD) is the host molecule and phenolphtalein (PP) is the guest has been studied in aqueous solution by UV-visible absorption spectroscopy. The decrease of temperature of beta-cyclodextrin-phenolphtalein system resulted in a decrease in absorbance of the UV-vis spectrum. Under favourable conditions (0.1 mM beta-CD, 30 microM PP) the termochromic effect is very significant (approximately =0.1 U of absorbance/10 degrees C). The formation constant of inclusion complex was determined at various temperatures (from 10 to 70 degrees C) using Scott's equation. The association constants (K11) for the binding in 0.02 M sodium carbonate (pH 10.5) at 10 and 70 degrees C are 7.44 and 0.26 x 10(4) M(-1) respectively. The stoichiometric ratio of investigated complex was found to be 1:1 on wide range of beta-cyclodextrin:phenolphtalein concentration ratio (from 0.8:1 to 427:1). Additionally, strong interaction between cyclodextrin and tetrahydrofuran (THF) was observed and the inhibitory effect of tetrahydrofurane on the association of beta-CD PP complex was studied. From linear Van't Hoff plots thermodynamic parameters such as: the change of enthalpy (deltaH(o)) and change of entropy (deltaS(o)) were estimated and interpreted. PMID- 9863956 TI - Inclusion complex formation of steroidal drugs with hydroxypropyl-beta cyclodextrin studied by charge-transfer chromatography. AB - The interaction between 17 steroidal drugs and hyroxypropyl-beta-cyclodextrin (HPbetaBCD) was determined by charge-transfer chromatography and the relative strength of interaction was calculated. HPbetaCD interacted with each steroidal drugs decreasing the hydrophobicity of the guest molecules. The relative strength of interaction considerably depended on the structure of the drug molecule. Hydrophobicity parameters of drugs significantly influenced the strength of interaction indicating the involvement of hydrophobic forces in the binding of drugs to HPbetaCD. The marked influence of HPbetaCD on the hydrophobicity of drugs suggests that this interaction may modify the biological properties (adsorption, uptake, half-life etc.) of drug-HPbetaCD complexes drug resulting in modified efficacy. PMID- 9863955 TI - Human alpha1-glycoprotein acid as chiral selector in the enantioseparation of midodrine and deglymidodrine racemates by HPLC. AB - Human alpha1-acid glycoprotein (alpha1-AGP) has been used as a chiral stationary phase (CSP) for the enantioseparation of midodrine and deglymidodrine racemates in the same HPLC run. The imobilized AGP resulted as the best chiral selector for the enantioresolution of two compounds. Due to the modification of alpha1-AGP characters as a result of changing the composition of the mobile phase, an attempt study of the watery mobile phase (ionic strength and pH of the buffer, nature and concentration of the organic modifier) allowed for an increase in the enantioselectivity of the chromatographic system and an optimization of the resolution base-line of both enantiomeric pairs. PMID- 9863957 TI - HPLC-fluorescence determination of unconjugated estrogens in pharmaceuticals. AB - A fluorimetric liquid chromatographic method (lambda(ex) = 280 nm; lambda(em) = 312 nm) was developed for measurements of unconjugated estrogens (estradiol and estriol) in pharmaceutical dosage forms using a reversed-phase column with water acetonitrile at different composition as mobile phase. The in vitro release profiles of three different estradiol transdermal therapeutic systems were determined through a medical-grade silicone rubber subdermal implant material membrane, using a modified Franz diffusion apparatus at 37 degrees C in presence of PEG 400. The HPLC method possesses advantages of rapidity, simplicity and accuracy. PMID- 9863958 TI - Determination of fluoxetine and norfluoxetine in human plasma by high-pressure liquid chromatography with fluorescence detection. AB - Fluoxetine is an atypical antidepressant drug, which selectively inhibits the neuronal reuptake of serotonin, and is widely used in the treatment of depressive disorders. The aim of this research is the development of an HPLC method with fluorescence detection for the monitoring of fluoxetine plasma levels. The determination requires no more than 250 microl of plasma, which undergo solid phase extraction (SPE), then are injected in the HPLC. For the analytical separation a reversed phase C8 column (150 x 4.6 mm I.D.) was used, while the mobile phase was a mixture of acetonitrile and water containing perchloric acid and tetramethylammonium perchlorate (flow rate: 1 ml min(-1)). The very low levels of analytes in plasma required the employment of a fluorescence detector (lambda(exc) = 230 nm, lambda(em)=290 nm), which also granted a good selectivity. Fluoxetine is revealed as a single peak at a retention time of 9.7 min, while norfluoxetine, the main metabolite of fluoxetine, is revealed at a retention time of 8.1 min. Linearity was obtained over the concentration range 8-200 ng ml(-1) for both substances. The method seems suitable, in accuracy and precision, for the determination of fluoxetine plasma levels of patients; furthermore, it is rapid and sensitive. PMID- 9863959 TI - Mass spectrometric characterization and HPLC determination of the main urinary metabolites of nimesulide in man. AB - A study was undertaken for the characterization and quantitative determination of the main urinary metabolites of the non-steroidal anti-inflammatory drug (NSAID) nimesulide (4-nitro-2-phenoxy-methanesulfonanilide) in man following single oral administration (200 mg). Urines were collected from six healthy volunteers at 12, 24, 48, 72 and 96 h post-administration and submitted to liquid liquid extraction before (free metabolites) and after enzymatic hydrolysis (conjugated metabolites). The structure of the metabolites, isolated by TLC separation, was elucidated by mass spectrometry (electron impact ionization) and confirmed by synthesis. Five metabolites were identified: they arise from hydroxylation to the phenoxy nucleus (M1 = hydroxynimesulide); reduction of the nitro group to an amino derivative (M2); concomitant hydroxylation and reduction (M3); N acetylation of the M2 (M4) and of the M3 (M5) metabolites. Quantitation was by reverse phase high performance liquid chromatography (Supelcosil LC-18 DB column; mobile phase: sodium phosphate buffer (pH 3.0, 50 mM)-acetonitrile (gradient elution); flow rate: 1 ml min(-1); UV detection, 230 nm), procedure which allows in a single chromatographic run the simultaneous determination of the unchanged drug and of its metabolites. The urinary excretion of the drug and metabolites (free + conjugated) in the overall 96 h-interval accounts for approximately 40% of the administered dose: 17.55 +/- 3.6% M1; 0.72 +/- 0.43% M2; 2.45 +/- 1.22% M3; 19.07 +/- 4.3% M5. The bulk of the metabolites was in conjugated form. Percentages excretion of the unchanged drug and of M4 metabolite were below 0.5%. The described method is suited to specifically and quantitatively measure nimesulide and metabolites in human urine with acceptable precision and accuracy. PMID- 9863960 TI - Simultaneous HPLC determination of multiple components in a commercial cosmetic cream. AB - A high-performance liquid chromatographic method for the simultaneous determination of magnesium ascorbyl phosphate (I), imidazolidinylurea (II), a mixture of methyl-(III), ethyl-(IV), propyl-(V), butyl-(VI) parabens dissolved in phenoxyethanol, and ascorbyl palmitate (VII), was studied by using a cyano-propyl column and a methanol gradient at 220 and 240 nm. Calibration curves were found to be linear in the 0.05-5 mg ml(-1) range (compounds I, II, VII) and 0.9-160 mg ml(-1) (compounds III-VI). Linear regression analysis of the data demonstrates the efficacy of the method in terms of precision and accuracy. An extraction method is developed and validated in order to apply this chromatographic method to a commercial cosmetic cream. The precision of this method, calculated as the relative standard deviation (RSD) of the recoveries (1.57-2.21%) was excellent for all compounds I-VII. PMID- 9863961 TI - Isolation of an antibacterial component from roasted coffee. AB - A coffee beverage obtained from instant dark coffee that had been previously shown to possess high antibacterial activity, was acidified (pH 2) and extracted with ethyl acetate. After alkalinization (pH 12) the aqueous phase was re extracted with the organic solvent. The acidic and basic extracts were evaporated to dryness and the aqueous phase freeze-dried. Residues were dissolved in sterile water and assayed for antibacterial activity against two reference bacteria (Staphylococcus aureus ATCC 25923 and Streptococcus mutans 9102). The acidic extract was found to be highly active and was separated by gel permeation chromatography (GPC) into five fractions. Fractions GPC4 and GPC5 were found to possess antibacterial activity: most of the activity was evident in fraction GPC5. These fractions were separated by RP-HPLC using a gradient elution with methanol water as mobile phase. Both GPC fractions gave an active subfraction with methanol-water (70:30, v/v). The experimental conditions used to separate the antibacterial compound that originates during the roasting process, indicate that it possesses low molecular mass (probably no more than 200 Da), weak acidic properties and an lambda(max) at 205 nm. The very small amount of this compound isolated from roasted coffee, indicates that it may be a very strong antibacterial agent. PMID- 9863962 TI - Comparison of GC-MS and TLC techniques for asarone isomers determination. AB - Two chromatographic methods (GC-MS and TLC) have been developed for separation and determination of alpha and beta asarone from essential oils and alcoholic extracts. The study has been performed on the Acorus calamus (I) and Asarum europaeum (II) essential oils of Romanian origin and the alcoholic extract of Acorus calcamus L (III) and it is a consequence of the International Boards exigency regarding the presence of beta asarone in food, beverages and pharmaceuticals. The isomers were determined using both internal and external standard methods. Both SIM and SCAN techniques were used and the results were compared regarding the chromatographic resolution and interference compounds. The method exhibits good repeatability and low detection limit but is expensive and time consuming. The two isomers concentrations are 5.2- 6.7 microg ml(-1) (I), 460-510 microg ml(-1) (II) and 2.7 5.7 microg ml (III) for alpha asarone and 91 98 microg ml(-1) (I), 24-29 microg ml(-1) (II) and 88 97 microg ml(-1) (III) for beta asarone. The TLC method was developed as an alternative for the GC method. The separation was performed on silica gel plates using toluene: ethyl acetate 8:2 as mobile phase. The evaluation of the chromatograms was made by densitometry using multiple wavelength. The sum of the two isomers are between 80-120 microg ml(-1) (I) and 127-145 microg ml(-1) (III) using spectrophotometric detection and between 73-93 microg ml(-1) (I) and 99-105 microg ml(-1) (III) using fluorimetric detection. The results of the two chromatographic methods were compared. Even the GC is more sensitive, mathematical computations for spots optimization and interference elimination could improves the TLC quality results. PMID- 9863963 TI - Determination of imidazole antimycotics in creams by supercritical fluid extraction and derivative UV spectroscopy. AB - A supercritical fluid extraction (SFE) method was developed for the isolation of imidazole antimycotic drugs (miconazole, econazole, clotrimazole and bifonazole) from cream preparations. The SFE process involved static (1 min) and dynamic (4 min) extraction steps using pure and 10% methanol modified carbon dioxide. The SFE step was then followed by derivative UV spectrophotometric analysis. The method proved to be suitable for quality control assays of the examined antimycotics in commercial cream formulations. PMID- 9863964 TI - Analysis of ceftazidime and related compounds by micellar electrokinetic chromatography. AB - A micellar electrokinetic chromatographic method for the separation and quantification of ceftazidime, its delta2-isomer and pyridine (two ceftazidime related impurities) was developed and validated. Optimised conditions were obtained using an electrolyte system consisting of 25 mM sodium tetraborate, at pH 9.2, and 75 mM sodium dodecylsulphate. A limit of detection of 0.2 microg ml( 1) and a limit of quantitation of 0.6 microg ml(-1) were estimated for pyridine and delta2-isomer: this means that levels of < 0.1% of pyridine and delta2-isomer in ceftazidime can be determined. Calibration curves for all analytes were linear over the studied ranges with correlation coefficients >0.999. Good reproducibility for migration times and corrected peak areas were achieved (RSD % 0.3 and 1.0, respectively). The results demonstrate that the method is reproducible, accurate and appropriate for ceftazidime assay in pharmaceutical samples. PMID- 9863965 TI - Determination and pharmacokinetics of acyclovir after ingestion of suspension form. AB - The study describes, simple, precise, sensitive and accurate HPLC assay with spectrofluorimetric detection for the determination of acyclovir in human plasma. The method was linear over a range 25 1200 ng ml(-1). The average yield in this method exceeded 80%. Limits of quantitation and detection were 25 and 10 ng ml( 1), respectively. On the basis of reported method, a single-dose of pharmacokinetics on 24 men, in two doses (200 and 400 mg) of acyclovir suspension has been investigated. Pharmacokinetic parameters obtained from both doses of the drug were compared. The linearity of acyclovir pharmacokinetics in the investigated dose ranges has been confirmed. PMID- 9863966 TI - Determination of anti-secalin antibodies in sera from coeliac patients by ELISA based assay. PMID- 9863967 TI - Use of sensitized luminescence of lanthanides in analysis of drugs. AB - The sensitization of the Tb(III) and Eu(III) ions luminescence by drugs, the pyrocatechol, naphthyridine and heterocyclic aromatic acid derivatives, were studied. It was shown that a result of intramolecular energy transfer from the ligand molecule to the lanthanide ion increases the luminescence intensity of the latter by 10(8)-10(10) times. The luminescence properties of the complexes in solutions were investigated. The highly sensitive methods for luminescence determination of dopegyt, levodopum, dophaminum, nevigramon, furosemidum and cinchophenum were developed. The detection limits of the drugs are 0.0005, 0.02, 0.5, 0.01, 0.05 and 0.1, respectively. PMID- 9863968 TI - Terbium chelates for fluorescence immunoassays. AB - The sensitization of terbium(III) ion luminescence in the presence of 1-ethyl-14 dihydro-6,7-methylenedioxy-4-oxoquinoline-3-carboxylic (oxolinic) acid was studied. The terbium label is bound to the antibody with diethylenetriamine pentaacetic acid anhydride (DTPAA). Optimum luminescence intensity is observed at pH 7.5 and the luminescence significantly increases in the presence of cationic surfactant, cetyltrimethylammonium bromide. The sensitivity of Tb(III) detection is 5 x 10(14) mol l(-1). This luminescence system is proposed for time-resolved fluoroimmunoassay. PMID- 9863969 TI - Development of a densitometric method for the determination of cephalexin as an alternative to the standard HPLC procedure. AB - A HPTLC-densitometric method was developed in order to obtain a reliable procedure for routine analysis of cephalexin in pharmaceutical formulations. Optimization of TLC conditions for the densitometric scanning was reached by eluting HPTLC silica gel plates in an horizontal developing chamber. Quantitation of cephalexin was performed in single beam reflectance mode by using a computer controlled densitometric scanner and applying a five-point calibration. A linear regression has been found in the 200-1000 ng range. The setup method is precise, reproducible and accurate. Recovery was also assessed by comparison with the HPLC USP XXIII alternate method. In this case HPTLC-densitometry appears worth of consideration as being relatively inexpensive and time-saving (up to 12 samples can be determined simultaneously in less than 15 min with a solvent consumption of about 15 ml). The results suggest that the proposed method may be used in place of HPLC for the routine quantitation of cephalexin in both pure and dosage forms. PMID- 9863970 TI - Study of the inclusion of gliclazide in alpha-cyclodextrin. PMID- 9863971 TI - Validation study of the conductometrical analysis. Application to the drug release studies from controlled release systems. PMID- 9863972 TI - The prevention of Rh haemolytic disease of the fetus and newborn--general background. PMID- 9863973 TI - The scientific basis of antenatal prophylaxis. AB - BACKGROUND: Standard post-delivery administration of anti-D, together with further anti-D for events known to result in fetomaternal haemorrhage (FMH) during pregnancy, has reduced the incidence of alloimmunisation in RhD women to 0.83-1.5% in the UK. Residual alloimmunisation occurs mainly for two reasons: i) failure to administer sufficient anti-D at the correct time after known at-risk events, either during pregnancy or at delivery; and ii) alloimmunisation during pregnancy as a result of 'silent' FMH. The RhD antigen is well developed by 6 weeks' gestation and the fetoplacental blood volume increases during pregnancy. Studies show that 3% of pregnant women have FMH in the first trimester, 12% in the second, and 45% in the third. Analysis of alloimmunisation in primigravidae clearly shows that on average, 90% are detectable after 28 weeks' gestation. Additional anti-D prophylaxis during the course of pregnancy, starting at 28 weeks, can reduce alloimmunisation to a minimum by protecting against occult FMH. The identification of intrapartum alloimmunisation as being the 'true' cause of alloimmunisation is best assessed by studying first pregnancies, and rigorous analysis of antenatal anti-D efficacy should preferably include observation in second pregnancies to avoid underestimation of alloimmunisation. Ideally there should be no exclusions, i.e. treating both arms of the study on an 'intention to treat' basis, otherwise there will be an overestimate of efficacy under routine practice conditions. Initial safety concerns about effects of antenatal anti-D on the fetus have not been confirmed in practice. OPTIONS: Although there is only one randomised controlled clinical trial (with small numbers) demonstrating a further reduction in alloimmunisation following antenatal administration of anti D meriting a grade A recommendation (see appendix II), the total body of evidence of efficiency is compelling. Whilst two doses of 300 microg are effective, this is no more so than the single dose in practice, and as it requires considerably more anti-D immunoglobulin, it is probably not cost effective. If a single dose is to be given, it is too late at 34 weeks, and 28 weeks is to be recommended. If divided doses are to be given at 28 and 34 weeks, 50 microg is insufficient, and 100 microg is recommended. Two dose regimes can be recommended, as follows: i) single dose of 300 microg at 28 weeks--the results of the single 300 microg dose in first pregnancies is limited to the Canadian study with observed reduction from 1.6% (45/2768) in concurrent nonrandomised controls to 0.18% (2/1086) in the treatment group. ii) two doses of 100 microg at 28 and 34 weeks--the two controlled studies give similar results in first pregnancies. A reduction in alloimmunisation is seen from 1.11% (4/360) in controls to <0.28% (0/362) in the treatment group in the French study , and from 0.95% (19/2000) in controls to 0.32% (4/1238) in the treatment group in the English study. Whilst anti-D is in limited supply, it is more cost effective to restrict antenatal prophylaxis to first pregnancies. It is also probable that a single dose of 250 microg (as used in Europe) will be as effective in practice as the 300 microg dose, given the limitations of the anti-D quantification assay, and the vial overfill introduced by manufacturers, but this has not been formally proven in clinical trials. The number of RhD deaths is now very low, even with standard postpartum prophylaxis, but there is a systematic underreporting in the UK, due to early fetal deaths being recorded as 'abortion' rather than as haemolytic disease of the newborn (HDN). There have been no systematic studies on the reduction in mortality observed with antepartum anti-D. Nevertheless, it is self-evident that if immunisation is largely prevented, then so will fetal morbidity and mortality. PMID- 9863974 TI - Economics of antenatal prophylaxis. PMID- 9863975 TI - Shoulder instability. Review of current trends in treatment. PMID- 9863976 TI - Atraumatic shoulder instability. Discussion of classification and results after capsular imbrication. AB - This study consists of 27 shoulders in 24 patients whose atraumatic shoulder instability was treated with the imbrication procedure described by Rockwood. Follow-up after 2 years or more showed the following results: 37% excellent, 30% good, 15% fair and 18% poor (Rowe score). No statistically significant differences in the results were observed between the dominant and non-dominant arm, nor in post-operative return to sports activities. Those with voluntary instability had more laxity at follow-up and tended to have poorer results. Those with unidirectional laxity had only good or excellent results and showed no post operative laxity. We found no other reliable indication of the results of surgery. The difficulties of classifying shoulder instability by such factors as trauma, laxity and direction of instability are discussed. Overall, we consider capsular imbrication to be a good treatment for involuntary atraumatic shoulder instability. PMID- 9863977 TI - Early arthroscopic treatment of primary traumatic anterior shoulder dislocation. A follow-up study. AB - This study evaluates the results of early arthroscopic Bankart repair in patients with primary traumatic anterior dislocation of the shoulder. The patients' age range was 17-34 years. Arthroscopic Bankart repair was performed within 12 days after the dislocation. First follow-up was at 18 months. According to Rowe's score, 11 patients (73%) were excellent, 3 (20%) were good and 1 (7%) was poor. The median external rotation deficit was 4 degrees in the adducted position. At a second follow-up at 34 months, two patients had redislocated. Both of these patients had severe generalized joint laxity. Another patient reported frequent subluxations. We conclude that in young patients with primary anterior traumatic shoulder dislocation, early arthroscopic Bankart repair implies a low recurrence rate and restores shoulder function to normal. Generalized joint laxity could indicate an increased risk for recurrent dislocation. PMID- 9863978 TI - Bioabsorbable tacks for arthroscopic treatment of recurrent anterior shoulder dislocation. AB - Seventy-eight patients (82 shoulders) with symptomatic, recurrent anterior post traumatic shoulder instability and Bankart lesions were operated on with bioabsorbable tacks (Suretac fixators). All the patients were followed by an independent observer, with a median follow-up period of 27 (21-63) months. The recurrence rate was 8/82 (10%). The median Rowe score was 93 (37-100) points. The median Constant score for the index shoulders was 90 (34-100) points, compared with 93 (80-100) points for 59 non-operated healthy shoulders from the same cohort (P=0.03). The external rotation in abduction was 93 (50-135) degrees compared with 105 (75-145) degrees for the control shoulders (P=0.0018). Arthroscopic shoulder stabilization using bioabsorbable Suretac fixators appears to produce reliable results if used in patients with post-traumatic shoulder instability and a Bankart lesion. PMID- 9863979 TI - Arthroscopic shoulder stabilization using transglenoid sutures. AB - The results of arthroscopic stabilization using multiple transglenoid sutures in 24 patients with posttraumatic recurrent anterior shoulder instability are presented with a minimum follow-up of 2 years. No serious complications were recorded. There were 2 recurrences. The remaining 22 patients had good or excellent results according to the modified Rowe score, with a median score of 89. The median value for loss of external rotation was 5 degrees. Seventeen patients were active in sports and 11 returned to the same sports at the same competitive level. PMID- 9863980 TI - Multidirectional hyperlaxity of the shoulder: results of treatment. AB - We have evaluated the results after rehabilitation and compared this to inferior capsular shift in patients with multidirectional hyperlaxity (MDH). The patients (n=35) experienced either instability and/or pain. We divided the patients into two groups: one group (group A) included patients with MDH and only pain but no symptoms of instability (n=6). All these patients had initial rehabilitation, followed by surgery in four cases. None of the patients with only pain (n=6) were satisfied after rehabilitation. Only two out of four were satisfied after surgery. In the second group (group B) were included patients who had MDH with both pain and instability. Six patients had surgery without prior rehabilitation and 20 patients had initial rehabilitation. Eight of these patients had surgical stabilization after unsuccessful rehabilitation. Less than half of the patients with MDH and instability (n=20) were satisfied after rehabilitation alone. Twelve of 14 patients were satisfied after surgery. We conclude that patients with MDH and only pain are difficult to treat. Patients with MDH and instability respond only moderately to the exercise program. Surgery, in combination with physiotherapy, should be the initial treatment when instability is the main symptom. PMID- 9863981 TI - Anterior cruciate ligament instability and reconstruction. Review of current trends in treatment. PMID- 9863982 TI - Postural control--a comparison between patients with chronic anterior cruciate ligament insufficiency and healthy individuals. AB - Postural control in the sagittal plane was evaluated in 22 patients with chronic anterior cruciate ligament (ACL) deficiency and the result was compared to that of a control group of 20 uninjured subjects. Measurement of the body sway was done on a fixed and sway-referenced force plate in both single-limb and two-limb stance, with the eyes open and closed, respectively. Further, an analysis of the postural reactions to perturbations backwards and forwards, respectively, was made in single-limb stance. The results demonstrated statistically significant deficits of the postural control in the patient group compared to the control group, but also within the patient group. There was a significantly higher body sway within the patient group when standing on a stable support surface on the injured limb than standing on the uninjured limb with the eyes open, but no difference with the eyes closed. When standing on a stable support surface, there was a significantly higher body sway in the patient group standing on the injured leg than in the control group, both with eyes open and closed. The patient group also showed a significantly impaired postural control compared to the control group when standing on the uninjured leg with the eyes closed. There was no difference between the groups in the two-limb stance. When standing on the sway referenced support surface, the patient group had a significantly larger body sway than the control group when the eyes were open, but there was no significant difference between the groups with the eyes closed. The measurement of the postural corrective responses to perturbations backwards and forwards showed that the reaction time measured from the initiation of the force plate translation, and the amplitude of the body sway was significantly greater in the patient group than in the control group. We conclude that patients with a continuing chronic ACL insufficiency several years after injury have an impaired postural control in the antero-posterior direction in single-limb stance on their injured leg. They also show a greater body sway and a prolonged reaction time when subjected to antero-posterior perturbations when standing on their injured leg. PMID- 9863983 TI - Knee injury and Osteoarthritis Outcome Score (KOOS)--validation of a Swedish version. AB - The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living. PMID- 9863984 TI - Bone mineral assessments in the calcaneus after anterior cruciate ligament injury. An investigation of 92 male patients before and two years after reconstruction or revision surgery. AB - The aim of this study was to assess the Bone Mineral Areal mass (BMA) in the calcaneus of male patients with unilateral anterior cruciate ligament (ACL) injury before and after reconstruction and to assess whether the BMA ratio or the BMA of the injured and uninjured side correlated with the level of activity, functional performance or the time period between the injury and the reconstruction. Ninety-two male patients with unilateral ACL injury were included in the study. The BMA was analysed immediately prior to surgery: a median of 11 (2-192) months after the injury in 30 patients aged 26 (15-41) years scheduled for primary ACL reconstruction (Group A). Forty-nine patients aged 29 (18-49) years had their BMA analysed a median of 24 (23-29) months after the primary ACL reconstruction (Group B). Thirteen patients aged 27 (21-39) years had their BMA analysed a median of 24 (20-45) months after ACL revision surgery (Group C). The median BMA ratio (injured side/uninjured side) was 96 (88-105)% in Group A, 96 (86-118)% in Group B and 95 (83-111)% in Group C. In all three groups, the median BMA value in the calcaneus was significantly lower on the injured side compared with the uninjured side (P=0.001, P=0.0003, P=0.01 in Groups A, B and C, respectively). The time period between the injury and the reconstruction neither correlated with the BMA ratio nor the BMA of the injured or the uninjured side in Group A. The level of activity as measured by the Tegner activity level and the functional performance as measured by the one-leg-hop quotient did not correlate with the BMA ratio in any of the groups. In the 49 patients with a primary ACL reconstruction (Group B), the post-operative Tegner activity level correlated with the BMA, on both the injured and uninjured side (P=0.0003, P=0.0005, respectively), when the BMA was assessed two years after the index operation. Male patients with unilateral ACL injury had a significantly lower BMA in the calcaneus on the injured side compared with the uninjured side before primary reconstruction, two years after primary reconstruction and two years after revision surgery. The time period between the injury and the index operation did not correlate with the BMA. A high level of activity correlated with the BMA on both the injured and the uninjured side two years after primary reconstruction. PMID- 9863985 TI - Bone graft fixation in anterior cruciate ligament reconstruction evaluated by radiostereometric analysis. AB - For most anterior cruciate ligament reconstruction techniques, the tensile strength of the ligament substitutes and the surgical methods of fixation have been examined. In the present study seven consecutive patients with a chronic anterior cruciate ligament deficiency, who were reconstructed with a bone patellar tendon-bone graft, were followed. The migration of the bone blocks was examined with radiostereometric analysis immediately postoperatively and after 3 and 12 months. The median migration of the bone blocks in the femur was 0.4 mm and in the tibia 0.4 mm. Clinical examination was undertaken preoperatively and one year postoperatively. We conclude that the use of interference screws seems to be a reliable method for fixation in anterior cruciate reconstruction and that healing of the bone blocks occurs during the first three postoperative months. PMID- 9863986 TI - NSIP, UIP, and the ABCs of idiopathic interstitial pneumonias. PMID- 9863987 TI - Measuring exhaled nitric oxide: not only a matter of how--but also why--should we do it? PMID- 9863988 TI - Detecting airways obstruction from the tidal flow profile. PMID- 9863990 TI - Micronodules and emphysema in coal mine dust or silica exposure: relation with lung function. AB - The aim of this study was to investigate the respective effects of micronodules and pulmonary emphysema, detected by computed tomography (CT), on lung function in workers exposed to silica and coal mine dust. Eighty-three subjects exposed to silica (n=35) or to coal mine dust (n=48), without progressive massive fibrosis, were investigated by high-resolution and conventional CT scans to detect micronodules and to quantify pulmonary emphysema by measuring the relative area of the lung with attenuation values lower than -950 Hounsfield units. Sixty-six (54.5%) subjects had evidence of micronodules on CT scans. Smokers had micronodules more rarely than nonsmokers. Significant correlations were found between the forced expiratory volume in one second (FEV(1); % predicted) (r= 0.41, p<0.001), FEV1/vital capacity (VC) (r=-0.61, p<0.001), diffusing capacity of the lung for carbon monoxide (DL,CO) (r=-0.36, p<0.001) and the extent of emphysema. No difference was demonstrated in the linear relationships between the extent of emphysema and the pulmonary function according to the type of exposure or the presence of micronodules on CT scans. This study suggests that micronodules detected by computed tomography have no influence, by themselves, on pulmonary function and that they should only be considered as a marker of exposure. PMID- 9863989 TI - Idiopathic nonspecific interstitial pneumonia/fibrosis: comparison with idiopathic pulmonary fibrosis and BOOP. AB - Based on past difficulties in clinically differentiating patients with idiopathic pulmonary fibrosis (IPF), bronchiolitis obliterans-organizing pneumonia (BOOP), and nonspecific interstitial pneumonia/fibrosis (NSIP), which all manifest clinically as interstitial lung disease, experience with pathologically confirmed examples of the three diseases was reviewed to compare clinical profiles and prognosis and to define NSIP more clearly. Thirty-one patients (15 males and 16 females) were pathologically identified as NSIP and subclassified into either the cellular (n=16) or fibrotic group (n=15). All 31 patients were clinically considered to be idiopathic NSIP cases. Patients with idiopathic BOOP (n=16) and IPF (n=64) were compared with the NSIP patients. Subacute presentation of interstitial lung disease characterized both idiopathic NSIP and idiopathic BOOP. NSIP patients showed volume loss on a chest radiograph (29.0%) and honeycombing on a computed tomography scan (25.8%); these features were not found in BOOP patients. Bronchoalveolar lavage lymphocytosis was characteristic of both BOOP and NSIP. Two subgroups of NSIP can be recognized histologically: patients in the fibrotic group had a less favourable outcome than those in the cellular group. BOOP and NSIP had a more favourable outcome than IPF. In conclusion, idiopathic nonspecific interstitial pneumonia can be differentiated from other types of idiopathic interstitial pneumonia, both pathologically and clinically. PMID- 9863991 TI - Bronchoscopy with transbronchial biopsies: measurement of bleeding volume and evaluation of the predictive value of coagulation tests. AB - The objectives of this study were to measure the bleeding volume associated with fibreoptic bronchoscopy with transbronchial biopsies (TBB), to correlate it with coagulation tests and to compare bleeding volume in patients with and without lung transplant. A total of 104 consecutive TBB in 51 different patients was evaluated prospectively. Before each procedure, haemoglobin, blood platelets, prothrombin time (PT), activated partial thromboplastin time (aPTT) and bleeding time were measured. During the procedure, lavage fluid and blood were collected by suction. The haemoglobin concentration of the mixture was measured and bleeding volume was calculated. Clinically significant bleeding was arbitrarily defined as >20 mL blood present in lavage fluid. The mean+/-SD bleeding volume was 7+/-10 mL with no statistically significant difference between transplanted and nontransplanted patients. In eight procedures (7.7%) the bleeding volume was >20 mL (range 22-61 mL). Prebiopsy values for blood platelet counts, PT and aPTT did not predict a bleeding tendency in any of the procedures in which significant bleeding occurred. No correlation was found between bleeding time and bleeding volume in the 17 procedures performed in patients with a prolonged bleeding time (> or =10 min). The bleeding associated with transbronchial biopsies was usually minor and quantitatively similar in patients with or without lung transplant. Coagulation tests could not predict clinically significant bleeding, which may occur in patients with normal coagulation test results. PMID- 9863992 TI - Two-year results after lung volume reduction surgery in alpha1-antitrypsin deficiency versus smoker's emphysema. AB - Lung volume reduction surgery (LVRS) improves exercise capacity and relieves dyspnoea in patients with smoker's emphysema (SE). It is unclear, however, whether LVRS similarly improves lung function in alpha1-antitrypsin-deficiency emphysema (alpha1 E). To address this question, this study prospectively compared the intermediate-term functional outcome in 12 consecutive patients with advanced alpha1E and 18 patients with SE who underwent bilateral LVRS. Before surgery there were no statistically significant differences between the two groups in the six-minute walking distance, dyspnoea score, respiratory mechanics or lung function data, except for the forced expiratory volume in one second, which was lower in the deficient group (24 versus 31% of the predicted value; p<0.05). In both groups, bilateral LRVS produced significant improvements in dyspnoea, the six-minute walking distance, lung function and respiratory mechanics. In the alpha1E group, the functional data, with the exception of the six-minute walking distance, returned to baseline at 6-12 months postoperation and showed further deterioration at 24 months. The functional status of the SE group remained significantly improved over this period. In conclusion, the functional improvements resulting from bilateral lung volume reduction surgery are sustained for at least 2 yrs in most patients with smoker's emphysema, but this type of surgery offers only short-term benefits for most patients with alpha1E. PMID- 9863993 TI - Cysteine proteinases and cystatin C in bronchoalveolar lavage fluid from subjects with subclinical emphysema. AB - This study examined the role of cysteine proteinases and their inhibitor in the development of emphysema in comparison with neutrophil elastase (NE) complexed with alpha1-protease inhibitor (NE-alpha1-PI), which was previously demonstrated to be increased in bronchoalveolar lavage (BAL) fluid from subjects with subclinical emphysema. Eight nonsmokers and 31 current smokers with (n=17) and without (n=14) emphysema, as evidenced by lung computed tomographic scans, were studied. The concentrations of immunologically detected cathepsin L and cystatin C, but not cathepsin B, were significantly increased in BAL fluid from the smokers with emphysema compared with those without emphysema, although the activity of cathepsin L, measured using a synthetic substrate and cathepsin L, released from cultured alveolar macrophages at 24 h, did not show any significant difference between the two groups. When comparison was made only for the subjects aged <60 yrs, the difference between the two groups disappeared for cathepsin L, but remained for NE-alpha1-PI. There was no significant correlation between the level of cathepsin L and that of NE-alpha1-PI in BAL fluid from the subjects with emphysema. In conclusion, increased levels of cathepsin L and cystatin C were demonstrated in bronchoalveolar lavage fluid from subjects with subclinical emphysema. However, the roles of cathepsin L and neutrophil elastase in the development of emphysema may vary between subjects and between the young and the old. PMID- 9863994 TI - Bronchial obstruction and exhaled nitric oxide response during exercise in cold air. AB - This study examines whether exhausting exercise in cold air induces bronchial obstruction and changes in exhaled [NO] and in exhaled NO output (V'NO). Thus, eight well-trained males performed two incremental exercise tests until exhaustion, followed by 5 min of recovery in temperate (22 degrees C) and cold ( 10 degrees C) environments, at random. At -10 degrees C, they were dressed in warm clothes. Ventilation (V'E), oxygen consumption (V'O2), carbon dioxide production, cardiac frequency (fC), and [NO] and V'NO were measured continuously. Before and after each test, the subjects' maximal expiratory flow-volume curves and peak expiratory flow, forced expiratory volume in one second (FEV1) and forced expiratory flow at 25 (FEF25), 50 (FEF50) and 75% (FEF75) of forced vital capacity were determined. At -10 degrees C, significant decreases in FEV1 and FEF75 were observed after exercise. At rest and at the same submaximal intensity, V'O2, V'E and fC did not differ significantly. At rest and up to approximately 50% peak V'O2, [NO] and V'NO values were lower at -10 degrees C than at 22 degrees C. Thereafter, and during recovery, the V'NO response became similar at both -10 and 22 degrees C. This study confirms that considerable hyperpnoea in cold air causes a detectable airway obstruction. This airway cooling also induces an initial decrease in the exhaled NO response. Since endogenous NO-production is involved in bronchial dilation, it cannot be excluded that this lack of production may favour the appearance of airway obstruction. PMID- 9863995 TI - Hypocapnia-induced contraction of porcine airway smooth muscle. AB - Hypocapnia constricts peripheral airways in vivo. This study investigated the role of airway smooth muscle in this phenomenon and the mechanism of hypocapnia induced contraction in vitro. Isometric tension, intracellular pH (pHi) and intracellular free calcium concentration ([Ca2+]i) were measured in porcine airway smooth muscles suspended in organ baths in the presence of 5% or 0% CO2. In tracheal strips precontracted with carbachol, hypocapnic challenge (0% CO2) produced increases in tension, pHi, and [Ca2+]i. In bronchial rings or tracheal strips precontracted with carbachol, nifedipine administered between consecutive contractions attenuated responses to hypocapnia (75+/-11% above carbachol precontracted tension before nifedipine versus 39+/-9% after nifedipine, n=7 bronchial rings, p<0.05). Neither indomethacin (5 microM), nordihydroguaiaretic acid (10 microM) nor phenidone (10 microM) significantly altered responses. These data suggest that enhanced Ca2+ influx through voltage-dependent Ca2+ channels of airway smooth muscle cells is important in airway responses to hypocapnia. PMID- 9863996 TI - Concurrent measurement of smooth muscle shortening, lumen narrowing and flow to acetylcholine in large and small porcine bronchi. AB - Models of airway function indicate that responsiveness (flow reduction) to bronchoconstrictor provocation depends on airway smooth muscle shortening and airway wall morphology. The contribution of these factors to the responsiveness of central and peripheral bronchi was assessed. Lumen flow was recorded in porcine perfused small (2 min i.d.) and large bronchial segments (6 mm i.d.). Lumen diameter was recorded in the same airways after inserting an endoscope. Smooth muscle shortening, relative wall area (WAr), smooth muscle and cartilage thickness and mucosal folds were measured morphometrically. The effect of acetylcholine (ACh; 10(-6)-10(-1) M) on functional measurements was determined by curve fitting. Maximum muscle shortening was 30% in small and 19% in large bronchi (p<0.01) and lumen narrowing was 49% and 39%, respectively. High doses of ACh stopped flow in small bronchi, but produced a plateau in large bronchi. Small airways were 250-times more sensitive to ACh than large airways, for all measurements. Smooth muscle and cartilage thickness and numbers of mucosal folds were greater in large than in small bronchi (p< or =0.01). Lumen narrowing and flow reduction were greater than predicted on the basis of muscle shortening and WAr (p<0.05). The structure of airways from the two groups was qualitatively similar, but responses were markedly different. Greater narrowing and flow responses of small bronchi were directly associated with smooth muscle responsiveness in situ. The results suggest that in vivo changes in airway wall shape or dimensions, or luminal secretions, exert a significant effect on airway flow, particularly in the small airways. PMID- 9863997 TI - Effect of a novel PACAP-27 analogue on muscarinic airway responsiveness in guinea pigs in vivo. AB - A recent study showed that the novel pituitary adenylate cyclase-activating peptide (PACAP)-27 analogue [Arg15,20,21,Leu17]-PACAP-27-Gly-Lys-Arg-NH2 causes sustained airway smooth muscle relaxation in vitro. This study examined whether this analogue also has bronchoprotective effects, by inhibiting muscarinic airway responsiveness in vivo. Total lung resistance was measured in anaesthetized, tracheostomized and ventilated guinea-pigs. Increasing doses of acetylcholine were given i.v. once before and thereafter repeatedly each hour after intratracheal instillation of either the PACAP-27 analogue or the clinical beta2 agonist bronchodilator salbutamol. Mean arterial blood pressure (MAP) was monitored to detect cardiovascular side-effects. Both the PACAP-27 analogue and salbutamol significantly attenuated the airway responsiveness to acetylcholine. The total inhibitory effect of the PACAP-27 analogue (350 nmol) corresponded to that of salbutamol (35 nmol). The inhibitory effect of salbutamol (35 nmol) peaked during the second hour and disappeared prior to 5 h after administration. In contrast, the corresponding effect of the analogue (350 nmol) gradually increased and peaked during the fifth hour after administration, whereas it did not fade during the observation period. Both the PACAP-27 analogue (350 nmol) and salbutamol (35 nmol) produced a transient decrease in MAP within 6 min after administration. In conclusion, the novel pituitary adenylate cyclase-activating peptide-27 analogue has bronchoprotective properties, by decreasing muscarinic airway responsiveness in guinea pigs in vivo. The time course of its effect is compatible with a more sustained duration of action compared with salbutamol. PMID- 9863998 TI - Neutrophil chemokines in bronchoalveolar lavage fluid and leukocyte-conditioned medium from nonsmokers and smokers. AB - Polymorphonuclear neutrophils (PMN) have been implicated in the pathogenesis of emphysema. The chemokines interleukin-8(IL-8), growth-related oncogene (GRO alpha) and extractable nuclear antigen (ENA)-78 may be involved in the increased numbers of PMN in smokers' airspaces. The levels of these cytokines in bronchoalveolar lavage fluid (BALF) and bronchoalveolar lavage leukocyte conditioned medium (LCM), along with BALF PMN numbers in 12 smokers who abstained for 12 h (chronic smoking) or continued to smoke until I h before study (acute smoking) and seven nonsmokers were compared. Neutrophils in BALF increased in acute (1.96+/-0.53%, 0.99+/-0.32x10(6) cells) compared with chronic smokers (0.59+/-0.25%, 0.61+/-0.24x10(6) cells, p<0.05 nonsmokers) and nonsmokers (0.79+/ 0.29%, 0.05+/-0.01x 10(6) cells, p<0.05). There were no differences in IL-8 or GRO-alpha in BALF between smokers and nonsmokers. ENA-78 levels were lower in smokers (p=0.006). There was no difference in IL-8, GRO-alpha or ENA-78 in LCM from unstimulated cells in smokers versus nonsmokers. After stimulation with lipopolysaccharide (LPS) 10 ng mL(-1), IL-8 release in acute smokers (p=0.04) and GRO-alpha release in smokers (p=0.009) were significantly higher than in nonsmokers. Following stimulation with LPS 100 ng.mL(-1), GRO-alpha release was higher in smokers (p=0.03) and increased further in acute smokers (p=0.02 versus nonsmokers, p=0.04 versus chronic smokers) and ENA-78 release increased in smokers (p=0.02 versus non-smokers). In conclusion, influx of polymorphonuclear neutrophils into smokers' airspaces is an acute phenomenon and neutrophil chemokine release from mixed bronchoalveolar lavage leukocytes is influenced by cigarette smoking and endotoxins. PMID- 9863999 TI - Rhinitis alone or rhinitis plus asthma: what makes the difference? AB - This paper examines the clinical expression of asthma in a group of patients displaying rhinitis according to age, sex, associated symptoms, smoking, familial history of asthma, atopy, type of sensitization to aeroallergens (pollens and/or indoor allergens), total serum immunoglobulin E (IgE), and blood eosinophils. A total of 117 adults with rhinitis were analysed on the basis of symptoms. Among them, 51 also displayed asthma, defined as a history of recurrent episodes of dyspnoea with a reversible airflow obstruction or a positive methacholine challenge. The logistic regression analysis carried out in a stepwise approach, combining several factors, showed that various parameters affected the risk of having asthma associated with rhinitis. A further analysis was made in 74 rhinitis patients comparing 42 subjects without nonallergic airway hyperresponsiveness (NAAH) to 32 patients with asthma and NAAH. Atopy, high total serum IgE levels, elevated blood eosinophil count and maternal asthma were associated with asthma. Furthermore, in atopic patients, pollen sensitization was more closely related to rhinitis alone, whereas sensitization to indoor allergens was a major determinant for the association of asthma with the symptoms of rhinitis. The same risk factors as those found in the clinical part of the study discriminated the patients with rhinitis without NAAH from those with rhinitis, asthma and NAAH. In conclusion, this study gives new insights into the relationships between asthma and rhinitis. PMID- 9864000 TI - Age-dependent inaccuracy of asthma death certification in Northern England, 1991 1992. AB - Asthma mortality appeared to increase two-fold in the UK from the mid- 1970s to the early 1990s, but there is evidence of inaccuracy in asthma death certification and so a region-wide investigation was undertaken to assess whether this recorded statistical trend might have been partly or wholly artefactual. A total of 35 respiratory physicians, distributed in panels of three, systemically reviewed the hospital and general practice records of 210 subjects with physician diagnosed asthma who died in 1991 and 1992. The death certificates indicated that asthma was considered to be the primary cause of death in 103 (group 1), a contributory cause in 70 (group 2) and not relevant in 37 (group 3). There was agreement within the panels that 43% of the subjects had probably never suffered from asthma. Discordance between the panels and the certifying physicians over the correct death certification category was high for group 1 (45% for those aged <65 yrs, 75% for those aged > or =65 yrs and 64% overall) and group 2 (67%), but much less for group 3 (22%). This study concludes that asthma death certification provides a markedly inaccurate picture of asthma mortality, particularly in elderly subjects. Thus, it is speculated that if the magnitude of this source of inaccuracy has increased over the last two decades, the apparent recent increase in asthma mortality may be largely artefactual. PMID- 9864001 TI - An inhaled steroid improves markers of airway inflammation in patients with mild asthma. AB - Airway inflammation can be demonstrated in mildly asthmatic patients who are not treated with inhaled steroids. Current guidelines recommend that inhaled steroids should be introduced in mild asthmatics who use an inhaled beta2-agonist more than once daily. It was postulated that inhaled steroids can have anti inflammatory effects in patients with even milder disease. The effect of 4 weeks of treatment with budesonide (800 microg twice daily by Turbohaler) was studied in 10 steroid-naive mildly asthmatic patients (forced expiratory volume in one second (FEV1) = 96+/-1.4% predicted) who required an inhaled beta2-agonist less than one puff daily, in a double-blind, placebo-controlled, crossover fashion. Spirometry, exhaled nitric oxide (NO), bronchial responsiveness (provocative concentration causing a 20% fall in FEV1 (PC20)), and sputum induction were performed before and after each treatment period. Following budesonide treatment, there were significant improvements in FEV1, and PC20, in association with a significant reduction in the percentage of eosinophils in induced sputum. Exhaled NO levels tended towards reduction, but the change was nonsignificant. There were also nonsignificant reductions in sputum eosinophil cationic protein and tumour necrosis factor-alpha levels. In conclusion inhaled budesonide can lead to improvements in noninvasive markers of airway inflammation, in association with a small improvement in lung function, even in mildly asthmatic patients who require an inhaled beta2-agonist less than once daily. This suggests a potential benefit of inhaled corticosteroids, even in relatively asymptomatic asthma. PMID- 9864002 TI - Perception of respiratory symptoms after methacholine-induced bronchoconstriction in a general population. AB - In an epidemiological study, methacholine-induced bronchoconstriction was used as a physical illustration of the sensations associated with asthma. The objective of this study was to assess whether familiarity with these sensations could be used as a measure of asthma prevalence. Eight hundred and seventy six subjects aged 20-44 yrs completed a respiratory questionnaire and a measurement of airway responsiveness (the provocative dose of methacholine causing a 20% fall in forced expiratory volume in one second (PD20)). Subjects were asked about their perception of, familiarity with and description of the respiratory sensations experienced at the time of maximal bronchoconstriction. The questionnaire-derived lifetime prevalences of wheeze, chest tightness and undue breathlessness were 43, 35 and 22% respectively. Asthma medication was used by 8% and the lifetime prevalence of diagnosed asthma was 12%. Quantifiable levels of airway responsiveness were measured in 34%, and airway responsiveness in the range considered to be consistent with untreated active asthma was present in 21%. Bronchoconstriction was perceived by 59%. Perception of bronchoconstriction was associated with the magnitude of bronchoconstriction, younger age, female sex and questionnaire-reported symptoms. Of subjects able to perceive bronchoconstriction, 58% reported previous experience of (familiarity with) the associated respiratory sensations. Familiarity with the sensations of bronchoconstriction was associated with questionnaire-reported symptoms, diagnosed asthma and increasing levels of airway responsiveness. There was poor agreement between the respiratory symptoms reported by questionnaire before the methacholine test and the words used to describe respiratory sensations induced by the test. Familiarity with the sensations of methacholine-induced bronchoconstriction has all the appropriate associations of a measure of asthma prevalence and may be a useful adjunct to symptom questionnaires and airway responsiveness measurements in epidemiological studies. A sizeable number of subjects can be identified with intermediate levels of airway responsiveness, who are able to perceive bronchoconstriction and are familiar with the sensations associated with it, yet who are not recognized to suffer from asthma. PMID- 9864003 TI - LFA-1 expression by blood eosinophils is increased in atopic asthmatic children and is involved in eosinophil locomotion. AB - Allergic asthma is characterized by eosinophil migration in the airways, which is strictly dependent on the expression of adhesion molecules. This study investigated whether the expression of adhesion molecules on eosinophils is increased and associated with disease activity in allergic asthma. Twenty atopic asthmatic (AA) subjects and nine controls were studied and the expression of lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18), Mac-1 (CD11b/CD18) and very late antigen-4 (VLA-4; CD49d/CD29) on blood eosinophils was evaluated by specific monoclonal antibody (Mab) staining and flow-cytometric analysis. Compared with controls, eosinophils from AA showed increased expression of LFA-1 (p<0.005), but not of Mac-1 or VLA-4 (p>0.1). In addition, LFA-1 expression correlated positively with blood eosinophil number (r=0.792, p<0.05), while no correlations were observed between Mac-1 or VLA-4 expression and blood eosinophil number. The migration of eosinophils through human umbilical vein endothelial cells with or without anti-LFA-1, Mac-1 and VLA-4-blocking Mab was studied. Compared with controls, eosinophils from AA showed increased migration toward C5a (p<0.01). Cell migration was totally inhibited by preincubating eosinophils with anti-LFA-1 (p<0.05), while anti-Mac-1 had no effect (p>0.1). Thus, the expression of lymphocyte function-associated antigen-1 by blood eosinophils is increased in atopic asthmatics and seems to modulate the enhanced eosinophil migration observed in allergic asthma. PMID- 9864004 TI - Clinical efficacy of low-dose inhaled budesonide once or twice daily in children with mild asthma not previously treated with steroids. AB - The aim of the present study was to examine the efficacy of low-dose inhaled budesonide (BUD) administered via Turbuhaler once or twice daily on symptoms, lung function and bronchial hyperreactivity in children with mild asthma. One hundred and sixty-three children (mean age 9.9 yrs, 56 females/107 males) with mild asthma (forced expiratory volume in one second (FEV1) 103% of predicted, morning peak expiratory flow (PEF) 87% pred, reversibility in FEV1 3%, fall in FEV1 after exercise 10.4% from pre-exercise value) and not previously treated with inhaled steroids, were included in a double-blind, randomized, parallel group study. After a two-week run-in period, the children received inhaled BUD 100 microg or 200 microg once daily in the morning, 100 microg twice daily or placebo for 12 weeks. Exercise and methacholine challenges were performed before and at the end of treatment. After 12 weeks of therapy, the fall in FEV1 after an exercise test was significantly less in all three BUD groups (43-5.1%) than in the placebo group (8.6%). Bronchial hyperreactivity to methacholine with the provocative dose causing a 20% fall in FEV1 decreased significantly in the BUD 100 microg twice-daily group compared with placebo (ratio at the end of treatment 156%). Changes in baseline lung function (FEV1 and PEF) were less marked than changes in bronchial responsiveness. In conclusion, low doses of inhaled budesonide, given once or twice daily, provided protection against exercise induced bronchoconstriction in children with mild asthma and near normal lung function. PMID- 9864005 TI - Urban-rural differences in questionnaire-derived markers of asthma in Kenyan school children. AB - Grade 4 Kenyan children attending 10 randomly selected public primary schools in Nairobi (urban) and the Muranga District (rural) were surveyed to establish the prevalence of symptom markers of asthma and to assess the impact of urbanization. A respiratory health and home environment questionnaire was administered at school to parents or guardians. The questionnaire response rates were 94.2% (568/ 603) for Nairobi and 89.6% (604/674) for Muranga. The prevalence rates for asthma, defined as "attacks of shortness of breath with wheeze", were 9.5% for urban and 3.0% for rural children (odds ratio (OR) urban versus rural: 3.42; 95% confidence interval (CI): -1.96-5.91). This urban-rural gradient persisted after adjusting for urban-rural differences in host factors (including duration of breastfeeding and family history of asthma and/or allergy), but was largely explained by urban-rural differences in environmental factors, including indoor animals, sharing a bedroom with a smoker, parental education, house ventilation and exposure to motor vehicle fumes en route to school (adjusted OR: 1.59; 95% CI: 0.70-3.55). Similar results were obtained for all other symptoms. These findings confirm the clinical impression that asthma is an important illness in Kenya and underline the need for the further study of environmental factors amenable to intervention, particularly in urban areas. PMID- 9864006 TI - Analysis of expiratory tidal flow patterns as a diagnostic tool in airflow obstruction. AB - An index obtained from tidal expiration, the ratio of time to peak tidal expiratory flow (tPTEF) to expiratory time (tE), discriminates between groups with and without airflow obstruction in infants and children and correlates with other measurements of airflow obstruction in adults. The aim of this study was to determine whether the diagnosis of airflow obstruction could be made from an analysis of the later part of the expiratory tidal flow time curve, i.e beyond the maximum flow. One hundred and eighteen patients attending the lung function laboratory with a putative diagnosis of airflow obstruction were studied. From uncoached tidal breathing, measurements were made of the average time constant of the respiratory system (Trs) and extrapolated volume (EV). Forced expiratory spirometry and whole-body plethysmography were performed. In this cross-sectional study, Trs correlated with inspiratory airways resistance and forced expiratory volume in one second (FEV1), according to the linear regression equations, airway resistance (Raw)=3.03 Trs+1.2, r=0.65, p<0.001, and FEV1% predicted = 87.8-23.7 Trs, r=058, p<0.001. EV correlated positively with overinflation, functional residual capacity (FRC) % pred = 152 EV+103, r=0.68, p<0.001. This study shows that there is a relationship between these measurements made from analysis of tidal breathing and recognized measurements of airflow obstruction and overinflation. PMID- 9864007 TI - Tidal expired airflow patterns in adults with airway obstruction. AB - Earlier studies have shown that time and flow indices derived from tidal expiratory flow patterns can be used to distinguish the severity of airway obstruction. This study was designed to address two aspects of tidal expiratory flow patterns: 1) how do expiratory flow patterns differ between subjects with normal and obstructed airways; and 2) can a sensitive index of airway obstruction be derived from these pattern differences? Tidal expiratory flow patterns from 66 adult subjects with varying degrees of airway obstructive disease with a forced expiratory volume in one second (FEV1) of 20-121% predicted were examined. In each subject, the expired flow pattern from each consecutive breath was scaled and then averaged together to create a single expired pattern. A detailed examination of the scaled flow patterns in 12 subjects (six with normal airways and six with airway obstruction) showed that the shape of the post-peak expiratory flow portion was different in the subjects with airway obstruction. A slope index, S, was derived from the scaled patterns and found to be sensitive to the severity of airway obstruction, correlating with FEV1 (% pred) with r2=0.74 (p<0.05, n=57). The S index also correlated (r2=0.36, p<0.05, n=47) with the functional residual capacity (FRC) (% pred) which was >100% in subjects with severe airway obstruction and lung overinflation. In subjects with normal airways, three further airflow patterns could be distinguished, which were different from the patterns seen in subjects with the severest airway obstruction. Scaled flow patterns from tidal expiration collected from uncoached subjects, can be used to derive an index of airway obstruction. PMID- 9864008 TI - Pulmonary function in sickle cell disease with or without acute chest syndrome. AB - Recurrent acute chest syndrome (ACS) has been suggested as a risk factor for chronic lung dysfunction in sickle cell disease. To investigate this hypothesis, lung function tests were performed in 49 sickle cell disease outpatients whose condition was stable, including 23 patients with a history of two to four episodes of ACS (ACS+) and 26 with no history of ACS (ACS-). The two groups were comparable regarding the sex ratio, body mass index, smoking history, physical characteristics, clinical history and usual lung function tests. Respiratory resistance (Rrs), measured using the forced oscillation technique, increased with the number of ACS episodes (r=0.55, p<0.0001) and a significant relationship was observed between Rrs as an independent variable and the expiratory flow rates at 25, 50 and 25-75% of the forced vital capacity as explanatory variables (r= 0.36, p<0.02; r=0.35, p<0.02; and r=0.4, p<0.006, respectively), with higher Rrs being associated with lower expiratory flow rates. The transfer factor (TL,CO) and transfer coefficient (KCO) for CO were significantly higher in the ACS+ group than in the ACS-group (TL,CO=84+/-4 versus 71+/-3%, p<0.004 and KCO=102+/-5 versus 90+/-3%, p<0.05, respectively). The data demonstrate that obstructive lung dysfunction is fairly common in sickle cell disease and suggest that recurrent acute chest syndrome may contribute specific obstructive defects. The increase in respiratory resistance associated with acute chest syndrome was accompanied by an increase in diffusion capacity, suggesting that it may have been related to an increase in lung blood volume. PMID- 9864009 TI - Lung function and exercise capacity in thalassaemia major. AB - Lung function abnormalities in thalassaemia major are various and complex; however, patients still die from cardiac lesions. This study aimed to investigate pulmonary and cardiac involvement at an early stage in thalassaemic patients and study their respective implications at rest and during exercise. Ten patients (five adults and five children) with thalassaemia major were investigated by echocardiography, lung function and exercise testing a few days after transfusion. All have had regular transfusions and chelation with deferoxamine and none had chronic pulmonary disease symptoms. Minor lung function abnormalities were found: two patients had moderate obstructive syndrome and two had a decreased carbon monoxide transfer factor. Hypoxaemia was never found at rest and no desaturation was observed at the end of exercise. Echocardiographic abnormalities were also moderate. Peak oxygen consumption (V'O2) was decreased in three adults and was lower in adults than children (means 27.7+/-4.6 and 41.1+/ 4.8 mL.kg(-1).min(-2) respectively). The V'O2/cardiac frequency slope was lower in adults than children (0.25+/-0.06 versus 0.42+/-0.10 mL.kg(-1).min(-2)), whereas end-exercise breathing reserve was >40% maximal voluntary ventilation for all patients. In conclusion, none of the patients had ventilatory limitations but older patients had cardiac limitations assessed by the relationship between oxygen consumption and cardiac frequency. Exercise testing may detect cardiac impairment in thalassaemia major earlier than investigations at rest. PMID- 9864010 TI - Immunolocalization of G(alpha)i-3 in foetal lung fibroblasts. AB - Guanosine triphosphate (GTP)-binding proteins or G proteins are involved in a wide variety of well-recognized signalling activities between cell surface receptors and effectors. The heterotrimeric G proteins have alpha,beta and gamma subunits organized in a trimeric structure. The aim of this study was to localize G(alpha)i-3, an important heterotrimeric G protein, in foetal lung cells. Using a foetal lung fibroblast cell line (RFL-6), the localization of G(alpha)i-3 was determined by immunofluorescence using a specific antibody to G(alpha)i-3, colocalization with a lectin known to bind the Golgi complex and Western blotting of RFL-6 cellular membrane proteins. This study identified G(alpha)i-3 on the Golgi membranes in rat foetal lung cells. Treatment with cycloheximide, to block protein synthesis, diminished the cytosolic distribution of the protein, but intense Golgi staining remained. G(alpha)i-3, therefore, appears to be part of the Golgi complex and not present transiently during protein synthesis. In the nonpolar foetal lung fibroblasts studied, the intracellular concentration of G(alpha)i-3 suggests a role for this protein in the intracellular trafficking and regulation of proteins needed for normal lung development. PMID- 9864011 TI - Alveolar lining fluid regulates mononuclear phagocyte 5-lipoxygenase metabolism. AB - The enzyme 5-lipoxygenase (5-LO) catalyses the synthesis of leukotrienes (LT), which are important in phagocytosis and killing of microorganisms. The alveolar macrophage (AM), the primary resident defender of the alveolar space, has a greater capacity for LT synthesis than its precursor, the peripheral blood monocyte (PBM). This study investigated whether the alveolar lining fluid (ALF) upregulates LT synthetic capacity in mononuclear phagocytes. Rat AM, peritoneal macrophages (PM) and ALF were obtained by lavage from pathogen-free animals. Human PBM were isolated from normal subjects. 5-LO metabolism and expression were measured with and without ALF. Rat ALF increased 5-LO metabolism (136.4+/-15.1% of control) in cultured PBM. This was associated with increased 5-LO activating protein (FLAP) (357+/-29.5 %), and 5-LO expression (188+/-31.3%). Culture of AM for 3 days resulted in a greater decrement in LTB4 synthesis (LTB4 15.4+/-6.9% of day 1) than in PM (54.7+/-8.3% of day 1), suggesting a greater dependence of AM 5 LO metabolism on ALF. 5-LO and FLAP expression decreased to a greater degree in AM than PM in culture. Furthermore, AM cultured with ALF maintained their LT synthetic capacity, FLAP and 5-LO expression compared with control cells cultured in medium alone. In conclusion, alveolar lining fluid increased 5-lipoxygenase metabolism in peripheral blood monocytes and maintained it in cultured alveolar macrophages, by a mechanism of increased 5-lipoxygenase and 5-lipoxygenase activating protein expression. This may boost host defence capabilities. PMID- 9864012 TI - A 50-kDa variant form of human surfactant protein D. AB - The dominant form of human surfactant protein D (SP-D) is a multimeric collagenous glycoprotein composed of monomeric subunits that have a molecular mass of 43 kDa under reducing conditions. However, in evaluating monoclonal antibodies to human SP-D, an additional monomeric subunit was identified with a reduced molecular mass of 50 kDa. This 50-kDa variant was detected in approximately half of the samples evaluated and was found in lavage fluid from normal subjects, patients with alveolar proteinosis or idiopathic pulmonary fibrosis and in amniotic fluid. This 50-kDa variant had the same amino-terminal sequence, amino acid composition and apparent size of the carboxy-terminal collagenase-resistant fragment (20 kDa) as the 43-kDa subunit. The major difference was in the amino-terminal portion of the molecule and was due to altered glycosylation, as determined by carbohydrate staining, chemical deglycosylation, treatment with N-glycanase and neuraminidase and reduced signals for threonine at positions 5, 9 and 10 during amino-terminal sequencing. After gel filtration chromatography, the 50-kDa form was not present in the high molecular weight fraction, which is commonly used in purification of SP-D, but was found only in the smaller molecular weight fraction of monomers and trimers of SP-D. In conclusion, the 50 kDa-form of surfactant protein D is produced by post-translational glycosylation and does not form higher ordered oligomers, but its precise physiological function remains to be determined. PMID- 9864013 TI - Long-term efficacy of dietary weight loss in sleep apnoea/hypopnoea syndrome. AB - Weight loss is associated with clinical improvement in sleep apnoea/ hypopnoea syndrome (SAHS). The aim of this study was to ascertain whether the therapeutic efficacy of dietary weight loss is maintained in the long-term. From a total of 216 overweight SAHS patients treated by only a weight reduction programme, 24 cured by this method (apnoea/hypopnoea index (AHI) at diagnosis 443+/-27.8, after weight loss 3+/-3.1) were re-evaluated after a mean (+/-SD) period of 94.3+/-27.4 months post-cure. No correlation was found between changes in AHI and body mass index experienced by each patient in the two phases of the study (diagnosis to cure and cure to long-term follow-up), r=0.29, p=0.156, demonstrating a marked intra-individual variability. Six of the 13 patients who maintained their weight presented recurrence of SAHS (AHI 40.5+/-24.1) as did eight of the 11 who had regained weight (AHI 44.3+/-23.). Weight maintenance was more frequent among patients who had continued to attend periodic appointments, 10/11 versus 3/13 (p<0.001). In conclusion, weight-loss efficacy is maintained in the long-term in some sleep apnoea/hypopnoea syndrome patients. This study indicates the need for periodic follow-up of these patients as a reinforcement for weight maintenance and for early detection of the reappearance of sleep apnoea/hypopnoea syndrome. PMID- 9864014 TI - The relationship between cardiac output and posthyperventilation hyperpnoea in patients with essential hypertension. AB - Posthyperventilation hyperpnoea (PHVH) is the progressive decline in minute ventilation (V'E) that follows abrupt cessation of voluntary hyperventilation. It has been hypothesized that the increase in cardiac output (CO) during hyperventilation could contribute to the duration of PHVH. This hypothesis was tested by measuring the duration of PHVH in patients with essential hypertension, in whom the increase in CO as a result of various stimuli is less pronounced. Twenty male hypertensives (mean arterial blood pressure+/-SEM: 178/ 107+/-3/1 mmHg), and 12 age-matched male healthy subjects were studied. The study consisted of three periods: control (5 min), voluntary hyperventilation (2 min), and recovery (3 min). V'E, CO, end-tidal CO2 and O2 tensions were measured, and the time constant (tau) of the V'E decay during recovery calculated. The V'E decay was faster in hypertensives (tau: 0-8.4 s) than in healthy subjects (tau: 12-59 s; p<0.01). During voluntary hyperventilation, CO increased to a lesser extent in hypertensives (6.8+0.7 L.min(-1)) than in healthy subjects (12.9+/-1.1 L.min(-1); p<0.01). In hypertensives, changes in CO during voluntary hyperventilation were significantly related to tau (r=0.646; n=20; p=0.002). The less pronounced rise in cardiac output during hyperventilation in hypertensives could account for the shorter duration of posthyperventilation hyperpnoea. PMID- 9864015 TI - Acute effects of inhaled nitric oxide in adult respiratory distress syndrome. AB - This study evaluated the dose-response effect of inhaled nitric oxide (NO) on gas exchange, haemodynamics, and respiratory mechanics in patients with adult respiratory distress syndrome (ARDS). Of 19 consecutive ARDS patients on mechanical ventilation, eight (42%) responded to a test of 10 parts per million (ppm) NO inhalation with a 25% increase in arterial oxygen tension (Pa,O2,) over the baseline value. The eight NO-responders were extensively studied during administration of seven inhaled NO doses: 0.5, 1, 5, 10, 20, 50 and 100 ppm. Pulmonary pressure and pulmonary vascular resistance exhibited a dose-dependent decrease at NO doses of 0.5-5 ppm, with a plateau at higher doses. At all doses, inhaled NO improved O2 exchange via a reduction in venous admixture. On average, the increase in Pa,O2, was maximal at 5 ppm NO. Some patients, however, exhibited maximal improvement in Pa,O2 at 100 ppm NO. In all patients, the increase in arterial O2 content was maximal at 5 ppm NO. The lack of further increase in arterial O2 content above 5 ppm partly depended on an NO-induced increase in methaemoglobin. Respiratory mechanics were not affected by NO inhalation. In conclusion, NO doses < or =5 ppm are effective for optimal treatment both of hypoxaemia and of pulmonary hypertension in adult respiratory distress syndrome. Although NO doses as high as 100 ppm may further increase arterial oxygen tension, this effect may not lead to an improvement in arterial O2 content, due to the NO-induced increase in methaemoglobin. It is important to consider the effect of NO not only on arterial oxygen tension, but also on arterial O2 content for correct management of inhaled nitric oxide therapy. PMID- 9864016 TI - Assessment of ventilation-perfusion mismatching in mechanically ventilated patients. AB - The multiple inert gas elimination technique (MIGET) is a robust tool to assess both ventilation-perfusion (V'A/Q') distributions and the role of extrapulmonary factors determining arterial oxygenation during spontaneous breathing and in mechanically ventilated patients. Mixed expired gas sampling used in the MIGET is most often obtained from a 10-L mixing box (10L-MB) placed in the expiratory side of the ventilator circuit. Consequently, a considerable increase in the compression volume (Vc) would be expected which, in turn, can give rise to potential errors in the estimation of the effective tidal volume delivered to the patient. The effects of the 10L-MB on the Vc were compared with those produced by a newly designed 1-L, mixing box (IL-MB). At a given peak pressure (Ppeak) within the ventilator circuit, the Vc generated by the 10L-MB was about six-times higher than that produced by the 1L-MB. At a Ppeak =50 cmH2O, the Vc were 377 mL (10L MB) and 67 mL (1L-MB) (p<0.001). In six patients, the mixed expired partial pressures of the six inert gases simultaneously collected from the two mixing boxes fell on the identity line. V'A/Q' distributions recovered using each of the two mixing boxes were equivalent. With the IL-MB, the effects of different positive end-expiratory pressure levels (0, 6 and 12 cmH2O) on Vc and arterial carbon dioxide tension were negligible. In conclusion, the new 1-L mixing box provides efficient gas mixing and substantially decreases the compression volume. It is, therefore, recommended when studies requiring mixed expired gas are performed in ventilated patients. PMID- 9864017 TI - Factor V Leiden is not common in patients diagnosed with primary pulmonary hypertension. AB - Substantial evidence suggests that thrombosis contributes to the pathogenesis of primary pulmonary hypertension (PPH). An abnormal factor V (factor V Leiden) may contribute to thrombosis in the pulmonary microcirculation of PPH patients. A point mutation in which adenine is substituted for guanine at nucleotide 1691 (1691A) alters factor V so that it resists cleavage by activated protein C. Heterozygosity for the 1691A mutation is more common (2-8%) in Caucasian Europeans and Americans than in Africans (1%) and Asians (<1%). The aim of the study was to examine the prevalence of the mutation that codes for factor V Leiden in individuals with PPH. We identified 42 Caucasians diagnosed with PPH. We extracted deoxyribonucleic acid (DNA) from whole blood and assayed DNA samples for the point mutation (1691 A) that codes for factor V Leiden. One out of 42 (2.4%; 95% confidence interval=0.1-12.6) Caucasians diagnosed with PPH was heterozygous for the normal 1691G and mutant 1691A allele. All 10 individuals with familial PPH were homozygous for the normal 1691G allele. The prevalence of heterozygosity for the 1691A allele and the normal 1691G allele does not differ from that observed in reference (control) populations. The low prevalence of the 1691A mutation among individuals diagnosed with primary pulmonary hypertension provides evidence that factor V Leiden does not contribute to the pathogenesis of the disease in most patients. PMID- 9864018 TI - Pulmonary hypertension in high-altitude chronic hypoxia: response to nifedipine. AB - Permanent residents at high altitude may develop excessive polycythaemia (H-Hb) and pulmonary hypertension, which often leads to cardiac failure. Inhibitors of calcium channels have been shown to reverse pulmonary hypertension in respiratory diseases and in primary pulmonary hypertension, but their efficiency has not been evaluated in high-altitude-induced pulmonary hypertension. Systolic pulmonary arterial pressure (Ppa) was studied by Doppler echocardiography, at rest and after sublingual nifedipine, in 31 asymptomatic residents at 3,600 m. Individuals were separated into two groups according to resting Ppa: a group with low Ppa (< or =4.7 kPa, n=17) and a group with high Ppa (>4.7 kPa, n=14). Individuals were also split into two groups according to haemoglobin (Hb) concentration: a normocythaemic (L-Hb) group ([Hb] < or =180 g.L(-1), n=17) and a H-Hb group ([Hb] >180 g.L.(-1), n=14). No significant difference in Ppa was observed between the L Hb and H-Hb groups. There was no correlation between [Hb] and Ppa. Nifedipine induced a decrease of >20% in Ppa in two-thirds of the subjects. This response was correlated with higher levels of basal Ppa (p<0.001) and was inversely correlated with age in the L-Hb group (p<0.05). Pulmonary vasoreactivity to nifedipine was independent of the degree of H-Hb. Pulmonary hypertension secondary to chronic altitude hypoxia may be reversible, despite a possible remodelling of the pulmonary arterioles. PMID- 9864019 TI - Measuring tidal volume and functional residual capacity change in sleeping infants using a volume displacement plethysmograph. AB - The noninvasive measurement of infant lung function during unsedated sleep in infants has been a long-standing objective in paediatric respiratory medicine. This note reports on the design and performance of a head-out volume-displacement plethysmograph (VDP) that overcomes some of the limitations of traditional lung function apparatus. The VDP comprises a rigid acrylic box with an integral water sealed spirometer and a novel neck seal. The bilayer neck seal is of variable compliance and is comfortable and simple to use. The spirometer permits volume resolution of 1.5 mL and a dynamic range in excess of 100 mL. The frequency response extends from 0-7 Hz. Spirometer inertance was measured as 0.0015 kPa.L( 1).s(-2), resistance 0.021 kPa.L(-1).s(-1) and box capacitance 0.18L.kPa(-1). Tidal volume, respiratory rate and changes in functional residual capacity can be recorded during unsedated rapid eye movement and nonrapid eye movement whilst monitoring with conventional polysomnographic methods. The head-out configuration allows additional instrumentation to be implemented with ease, avoids facial stimulation and allows unimpeded access to the upper airway. A polysomnograph illustrating the limitations of respiratory inductance plethysmography signals and typical changes in functional residual capacity are shown. PMID- 9864020 TI - Single-breath method for assessing the viscoelastic properties of the respiratory system. AB - In order to explain the time dependency of resistance and elastance of the respiratory system, a linear viscoelastic model (Maxwell body) has been proposed. In this model the maximal viscoelastic pressure (Pvisc.max) developed within the tissues of the lung and chest wall at the end of a constant-flow (V') inflation of a given time (tI) is given by: Pvisc,max = R2V'(1-e(-tI/tau2), where R2 and tau2 are, respectively, the resistance and time constant of the Maxwell body. After rapid airway occlusion at t1, tracheal pressure (Ptr) decays according to the following function: Ptr(t) = Pvisc(t) + Prs,st = Pvisc,max(etocc/tau2)+ Prs,st, where tocc/is time after occlusion and Prs,st is static re-coil pressure of the respiratory system. By fitting Ptr after occlusion to this equation, tau2 and Pvisc,max are obtained. Using these values, together with the V' and tI pertaining to the constant-flow inflation preceding the occlusion, R2 can be calculated from the former equation. Thus, from a single breath, the constants tau2, R2 and E2 (R2/tau2) can be obtained. This method was used in 10 normal anaesthetized, paralysed, mechanically ventilated subjects and six patients with acute lung injury. The results were reproducible in repeated tests and similar to those obtained from the same subjects and patients with the time-consuming isoflow, multiple-breath method described previously. PMID- 9864021 TI - Seasonal clustering of sarcoidosis presenting with erythema nodosum. AB - Seasonal clustering of sarcoidosis presenting with erythema nodosum (EN) has previously been reported only in the northern hemisphere. Of 59 patients presenting to a single centre in New Zealand with a new diagnosis of sarcoidosis, 21 had EN and three more had acute arthralgia without EN. These patients were compared with the rest of the cohort. The patients with EN or arthralgia alone presented exclusively between April and December, with peak clustering in the spring months of August, September and October (p<0.001, Fisher's exact test). This cohort was more likely to have a stage I chest radiograph and to be female (p<0.05), but there were no other differences between the groups. This is the first report of seasonal clustering in the southern hemisphere suggesting a common environmental trigger in the aetiology of sarcoidosis. PMID- 9864022 TI - Neutrophil serine proteinases and defensins in chronic obstructive pulmonary disease: effects on pulmonary epithelium. AB - Neutrophils have the capacity to accumulate in high numbers in the lung during infection and inflammation. Because they play an important role in host defence against infection, but may also cause tissue injury, these cells are thought to be involved in the pathogenesis of various inflammatory lung disorders, including chronic bronchitis and chronic obstructive pulmonary disease. Neutrophil products that may mediate tissue injury at sites of neutrophil-dominated inflammation include the neutrophil serine proteinases elastase, cathepsin G and proteinase 3, and the nonenzymatic defensins. One of the targets of the neutrophil is the lung epithelium, and in vitro studies have revealed that both the serine proteinases and neutrophil defensins markedly affect the integrity of the epithelial layer, decrease the frequency of ciliary beat, increase the secretion of mucus, and induce the synthesis of epithelium-derived mediators that may influence the amplification and resolution of neutrophil-dominated inflammation. Both neutrophil elastase and defensins induce the release of the neutrophil chemoattractant chemokine interleukin-8 from respiratory epithelial cells. The alpha1-proteinase inhibitor (alpha1-PI) is a well-characterized inhibitor of neutrophil elastase, that also blocks the cytotoxic and stimulatory activity of defensins towards epithelial cells. The elastase inhibitory activity of alpha1-PI is also abrogated by the binding of defensins to this inhibitor. Incubation of epithelial cells with neutrophil defensins in combination with either elastase or cathepsin G resulted in decreased effects on the epithelial cells compared with those observed when the cells were incubated with defensins, elastase or cathepsin G separately. These results suggest that neutrophil defensins and serine proteinases cause injury and stimulate epithelial cells to produce chemokines that attract more neutrophils to the site of inflammation. The effects of neutrophil defensins and serine proteinases on epithelial cells appear to be restricted by proteinase inhibitors and by inhibitory interactions between these sets of neutrophil granule proteins. PMID- 9864023 TI - Difficult asthma. AB - Asthma is usually easy to manage, but approximately 5% of patients are not controlled even on high doses of inhaled corticosteroids. It is important to assess these patients carefully in order to identify whether there are any correctable factors that may contribute to their poor control. It is critical to make a diagnosis of asthma and to exclude other airway diseases, particularly chronic obstructive pulmonary disease (COPD), and vocal cord dysfunction ("pseudo asthma"). Poor adherence to therapy, particularly inhaled corticosteroids, is a common reason for a poor response. There may be unidentified exacerbating factors, including unrecognized allergens, occupational sensitizers, dietary additives, drugs, gastro-oesophageal reflux, upper airway disease, or other systemic diseases, that need to be identified and avoided or treated. Psychological factors may be important in some patients, but it is difficult to know whether these are causal or secondary to troublesome disease. Some patients have instability of their asthma, with resistant nocturnal asthma, premenstrual exacerbations or chaotic and unpredictable instability (brittle asthma). A few patients are completely resistant to corticosteroids, but more patients are relatively resistant and require relatively high doses of corticosteroids to control their symptoms (steroid-dependent). Some patients develop progressive loss of lung function, as in patients with COPD. Management of patients with difficult asthma should be supervised by a respiratory specialist and should involve careful assessment to confirm a diagnosis of asthma, identification and treatment of exacerbating factors, particularly allergens, and recording of peak expiratory flow patterns. A period of hospital admission may be the best way to assess and manage these patients. Treatment involves optimizing corticosteroids therapy, assessing additional controllers such as long-acting inhaled or subcutaneous beta2-agonists or subcutaneous, theophylline and antileukotrienes. In some patients, the use of immunosuppressive treatments may reduce steroid requirements, although these treatments are rarely effective and have side effects. In the future, the nonsteroid anti-inflammatory treatments now in development may be useful in these patients. PMID- 9864024 TI - Assessing the effect of deep inhalation on airway calibre: a novel approach to lung function in bronchial asthma and COPD. AB - Bronchoconstriction in bronchial asthma and chronic obstructive pulmonary disease (COPD) may be due to decreased airway calibre and/or to the inability of the airways to distend after a deep inhalation (DI). The purpose of this review is to discuss the physiological and clinical relevance of this latter mechanism. During induced constriction, DI shows remarkable bronchodilatation in normal subjects, but a blunted or null effect in asthmatics. In contrast, during spontaneous bronchospasm DI tends to decrease airway calibre. From a functional point of view, airway inflammation, remodelling, and peripheral bronchoconstriction could prevent airway smooth muscle from stretching. Therapeutic intervention improving lung function may change the response to DI. For example, bronchodilators allow expiratory airflow before DI to increase more than after DI, because of decreased bronchial hysteresis. This suggest that bronchodilation might be systematically underestimated from parameters derived from maximal expiratory manoeuvres. Inhaled corticosteroids tend to increase the dilator effect of DI, likely due to decreased bronchial and peribronchial oedema. In conclusion, measuring the effects of deep inhalation on lung function is an easy and simple test able to evaluate the structural changes occurring in the airways and to monitor the effectiveness of therapy. PMID- 9864025 TI - Systemic vasculitis in a patient with small-cell neuroendocrine bronchial cancer. AB - A 75-yr-old male hospitalized for vascular purpura with joint pain had a medical history of polymyalgia rheumatica. A generalized oedematous syndrome occurred and the patient also presented with haemoptysis and complained of transient paraesthesia of the hands and feet. Renal biopsy showed lesions of focal segmental proliferative glomerulonephritis associated with a few cellular crescents. Lung biopsy showed small-cell neuroendocrine carcinoma. After the first course of chemotherapy signs of vasculitis disappeared. Small-cell neuroendocrine carcinomas, which represent 25% of all lung cancers, have numerous paraneoplastic (especially neurological) extrapulmonary manifestations. Disseminated vasculitis has never been described with this type of cancer, whereas nonsmall-cell carcinomas are associated essentially with cutaneous vasculitis or purpura rheumatica. In the case reported here, anticancer chemotherapy allowed vasculitic manifestations to be treated. PMID- 9864026 TI - Fibrous pleural tumour producing 171 litres of transudate. AB - Localized fibrous tumours of the pleura (or localized benign mesothelioma) are rare, and in most cases, asymptomatic. This report describes a 48 yr old female with a right-sided fibrous pleural tumour, which produced 171 L of transudate before a correct diagnosis was reached. The tumour was surgically removed and the transudation stopped immediately. PMID- 9864027 TI - Chylothorax or leakage of total parenteral nutrition? AB - The diagnosis chylothorax is based on a chemical analysis of the pleural effusion. According to the literature, this analysis can be rather straightforward, comprising measurements of triglycerides, chylomicrons, and cholesterol. In this report we present an autopsy case that alerted us to interpret these results critically. Although the laboratory tests of the pleural effusion in this patient with parenteral nutrition suggested chylothorax, additional tests (potassium (11.3 mmol.L(-1)) and glucose (128 mmol.L(-1)) proved otherwise. Comparison of the pleural effusion analysis and the content of the parenteral nutrition led to the final conclusion that the effusion was due to a leakage of parenteral nutrition instead of chylothorax. We therefore suggest adding glucose and potassium measurements to the biochemical work-up of a patient under suspicion of chylothorax. PMID- 9864028 TI - Diagnosis and treatment of pulmonary tuberculosis. PMID- 9864029 TI - HLA restriction patterns of gliadin- and astrovirus-specific CD4+ T cells isolated in parallel from the small intestine of celiac disease patients. AB - Celiac disease is a common HLA-DQ2-associated enteropathy caused by an abnormal T cell-mediated immune response to ingested wheat gliadin proteins. We have previously isolated in situ activated mucosal T cells from celiac disease patients and demonstrated that these T cells were gliadin specific and predominantly DQ2 restricted. In contrast to this, gliadin-specific T cells isolated from peripheral blood display a variable HLA restriction pattern, thereby indicating that the skewed DQ restriction of T cells resident in the celiac lesions could be dictated by a preference for DQ-mediated antigen presentation in the mucosa of CD patients. To address this, we analyzed the HLA restriction of T cells recognizing astrovirus, a common gastroentetitis virus, isolated from intestinal mucosa of six celiac disease patients. As an internal control, gliadin-specific T cells were isolated and analyzed in parallel. The gliadin-specific mucosal T cells were marked in their DQ2 restriction, whereas the parallel astrovirus-specific T cells were predominantly restricted by DR molecules. Our data indicate that the repertoire of T cells present in celiac lesions is determined by the priming antigen(s) and not by a skewing in the expression of functional HLA class II isotypes in the disease affected small intestinal mucosa. PMID- 9864030 TI - A single amino acid change makes the peptide specificity of B*3910 unrelated to B*3901 and closer to a group of HLA-B proteins including the malaria-protecting allotype HLA-B53. AB - HLA-B*3910, which has only been found in African and African American individuals, differs from B*3901 by the single amino acid change of Cys67 to Tyr67. Sequence analysis of the B*3910-bound peptide pool and of several individual ligands revealed that this subtype has strong preference for peptides with Pro2. This is in contrast with the preference of B*3901 for peptides with basic residues (Arg and His) at this position, and indicates that the single amino acid substitution between B*3910 and B*3901 totally changes the repertoire of bound peptides. This is presumably due to the significant decrease in the size of the B pocket, and to its increased hydrophobicity, since Tyr67 takes part in this pocket. B*3910 is similar to various other class I proteins in its preference for peptides with Pro2 and nonpolar C-terminal residues, including HLA B53, an antigen associated with protection against severe malaria. The role of these two motifs as major peptidic anchors suggests that B*3910 and HLA-B53 may bind common peptides. PMID- 9864031 TI - Monoclonal antibody NU-B1 reacts with novel antigen on human B cells in mantle and marginal zones distinct from known CD molecules. AB - To analyze the cellular antigens of human B-cell lineage, a monoclonal antibody, NU-B1, was raised using the acute lymphoblastic leukemia (ALL) cell line NALM-16 as the immunogen. NU-B1 reacted with 7.7+/-3.9% of the healthy adult peripheral blood mononuclear cells but not with neutrophils, monocytes, red blood cells or thymocytes. In order to distinguish the reaction specificity of NU-B1, two-color immunofluorescence staining using tonsillar cells was performed, and it was demonstrated that NU-B1-positive cells coexpressed CD20, which is a representative B-cell antigen. The expression of NU-B1 was highly restricted to cells of B-cell lineage when a panel of hematopoietic cell lines was examined. In a pathoimmunohistological study using human lymph node tissue, NU-B1-positive cells were localized in the mantle and marginal zones. In a clinical study, NU-B1 reacted specifically with leukemias/lymphomas of B-cell lineage: all 43 cases of ALL including common ALL and biphenotypic leukemia, all 4 cases of B-cell ALL, 6/7 B-cell type malignant lymphomas and 2/4 B-cell chronic lymphocytic leukemias. NU-B1 did not react with multiple myeloma, T-cell or myeloid leukemias/lymphomas. Immunoprecipitation of NU-B1 revealed two clear bands at 50 kDa and 42 kDa under either reducing or nonreducing conditions. Although anti-IgM treatment induced dramatic down modulation of CD79b, the NU-B1 antigen was also down modulated, but only slightly. However, crosslinking of NU-B1 did not induce tyrosine phosphorylation of intracellular proteins or the mobilization of calcium in NALM 16. The present results revealed that the antigenic determinant recognized by NU B1 is not surface immunoglobulin chains, HLA-DR, a receptor for C3, Fc for immunoglobulin chains or any known CD molecule. We conclude that monoclonal antibody NU-B1 recognizes a novel human B-cell restricted antigen distinct from known CD molecules, and that it is a useful antibody in the immunophenotyping and classification of leukemias/lymphomas. PMID- 9864032 TI - Successful in vitro immortalization of human intestinal mucosal lymphocytes with Herpesvirus saimiri. AB - Mucosal intestinal lymphocytes form the first immune-cell line of defense in the intestine. Several methodologies, most of them cumbersome and time consuming, have been used to obtain T-cell clones to unveil their physiological role. In the present work we take advantage of the recently described technique of transformation of T lymphocytes using Herpesvirus saimiri to show that it is possible to immortalize intestinal T-cell lines derived from healthy and diseased colonic samples and thence easily obtain in vitro intestinal T-cell lines as a model for physiopathological studies. Intestinal samples were obtained by colonoscopy and digested with dispase and collagenase. Mucosal lymphocytes (assessed by the expression of the CD3 and CD103 markers) were isolated using a Percoll gradient centrifugation and transformed with Herpesvirus saimiri. Sustained growth was observed 3 months later, showing that the cells were successfully transformed, a finding further confirmed by PCR. All cell lines were CD8+TcRalphabeta+ and HLA-DR+. CD25 was expressed on 1% of Crohn's disease derived cells and on 25% of cells derived from patients with ulcerative colitis. CD80 expression was found on 80-90% of the cells. These immortal cell lines of intestinal origin may be useful in future experiments aimed at elucidating the role of mucosal lymphocytes in health and disease. PMID- 9864033 TI - Donor-recipient sharing of HLA class II alleles predicts earlier recurrence and accelerated progression of hepatitis C following liver transplantation. AB - Both direct viral cytopathic effects and host immune responses appear to be important in the pathogenesis of hepatitis C virus (HCV) infection. Liver transplantation provides a means to explore the role of the immune system in the development of HCV-related liver damage through comparing the natural history of HCV in patients with different degrees of donor-recipient human leukocyte antigen (HLA) matching. We evaluated 36 patients with recurrent hepatitis C viremia following liver transplantation to determine whether hepatocellular injury or progression to bridging fibrosis occur more rapidly when donor and recipients share HLA alleles. HLA typing for the HLA-A and HLA-B loci was performed by serological techniques and PCR-based oligotyping was used to type alleles of the DRB1, DRB3, DQA1, and DQB1 loci. A median of eight liver biopsies, obtained during a median follow-up of 36 months, were reviewed per patient. Donor recipient sharing of alleles of HLA-DQB1 or DRB1 was associated with more rapid development of recurrent hepatitis by univariate analysis (chi2=5.7, P=0.02 and chi2=5.54, P=0.02 respectively). However, only sharing of HLA-DRB1 alleles was identified as an independent predictor of reduced time to recurrent histologic injury by multivariate analysis (chi2 =5.74, P=0.02). Furthermore, sharing of HLA DRB3 and histologic evidence of rejection were associated with more rapid progression to bridging fibrosis both by univariate methods (chi2=4.12, P=0.04 and chi2=4.66, P=0.03 respectively), and by multivariate analysis (chi2=13.01, P=0.001). These findings suggest that HLA class II-restricted immune responses may contribute to the pathogenesis of HCV-related liver injury in liver transplant recipients. PMID- 9864034 TI - HLA class I and class II of the Nivkhi, an indigenous population carrying HTLV-I in Sakhalin, Far Eastern Russia. AB - The Nivkhi are a native people isolated in the Nogliki region of Sakhalin, Far East Russia, where our group recently recognized human T-cell lymphoma virus type I (HTLV-I) infection. In order to trace the Nivkhi's ethnic background and the HTLV-I carriers, we investigated HLA class I and II allele types of 53 Nivkhi (including four HTLV-I carriers). Major HLA class I alleles of the Nivkhi were A*24, A*02, B*40, B*48, B*27, B*35 with allele frequencies similar to the Orochon, a native people isolated in Northeast China. Major Nivkhi class II alleles were DRB1*0901, DRB1*1401, DRB1*1201, DRB1*1106 with allele frequencies similar to the Ainu in Hokkaido, Japan, but dissimilar to other Asian Mongoloids, including the general Japanese population. The same HLA class I and II allele frequencies are found in both Nivkhi HTLV-I carriers and the background population. A dendrogram of HLA class I alleles showed that the Nivkhi were closely related to the Orochon and Yakut, and remotely related to the Japanese, Ainu and other Asian Mongoloids. Interestingly, the Nivkhi were intermediately related to the Amerindians (Inuit, Tlingit and Andeans), a relationship closer than to the Japanese and Asian Mongoloids. These results suggested the Nivkhi might be related to some genetic group of Northeast Asian Mongoloids like the Orochon and Yakut, being infected with HTLV-I in the distant past before diverging into the current major Mongoloid ethnic groups. PMID- 9864035 TI - HLA-B35 frequency variations correlate with malaria infection in Sardinia. AB - The hypothesis of a possible selective role of malaria in HLA allele frequency variations was investigated in Sardinia by typing completely 1,039 individuals for HLA: 536 from six lowland villages exposed to malaria until 1948, and 503 from six highland villages with no history of malaria. Another 1,928 individuals from 136 villages scattered all over the island were studied to establish if the HLA allele frequencies among villages correlated with the malaria incidence and/or altitude above sea level. Only the HLA-B35 allele yielded significantly higher frequencies in the lowland versus the highland villages (P<1 x 10(-5)). The observed B35 variance was 9.5 times higher than expected in the absence of selection, showing an adaptive origin. The highly significant positive correlation found between HLA-B35 frequency and malaria in 136 villages suggests that malaria has been the selective factor for HLA-B35 in Sardinia. PMID- 9864036 TI - Fluorotyping of HLA-A by sequence-specific priming and fluorogenic probing. AB - The aim of our study was to develop a fluorotyping strategy for the HLA-A locus. In contrast to conventional sequence-specific primed PCR (PCR-SSP), which is based on an agarose gel electrophoresis, fluorotyping eliminates the drawback of low sample throughput, low potential for automation and problems related to contamination. Additionally, fluorotyping is capable of delivering quantitative results depending on the system set-up. The fluorescence-based PCR-SSP assay relies on target-specific and individually labeled fluorogenic probes allowing a simultaneous and differential detection of the specific HLA and the internal control product. The probe used to detect the HLA-A specific amplicons was labeled at its 5' end with 6-carboxyfluorescein (FAM) as the reporter and at its 3' end with 6-carboxy-tetramethylrhodamine (TAMRA) as the quencher. The probe hybridized within the 2nd intron to a conserved region which was found to be identical in all HLA-A alleles and was covered by all primer mixes. During successful PCR the cleavage of the FAM-labeled probe through the 5'-3' exonuclease activity of the Taq DNA-polymerase led to an interruption of the TAMRA-mediated quenching effect and generated a significant increase of the FAM fluorescence. The specific HLA-A typing information was based on the FAM fluorescence released by 24 individual PCR primer mixes. The internal control amplicon was detected with a tetrachloro-6-carboxyfluorescein-TAMRA-labeled fluorogenic probe. Since the HLA-A amplicons had to include the 2nd intron as the target for the fluorogenic probe, the sequence motifs which could be used as priming sites were limited. Therefore, some primer mixes covered more than one specificity resulting in ambiguous amplification patterns in 31 of 231 possible allele or group combinations of HLA-A1-A80. These ambiguities, which all involved the inability to discriminate a particular heterozygous genotype from a homozygous genotype, may be resolved by the quantitative data revealed by fluorotyping. This feature is also helpful to detect new alleles which are not amplified by the current primer mixes. PMID- 9864037 TI - Distribution of insulin-dependent diabetes mellitus (IDDM)-related HLA alleles correlates with the difference in IDDM incidence in four populations of the Eastern Baltic region. AB - The high incidence of insulin-dependent diabetes mellitus (IDDM) in Finland contrasts strikingly with the low rates in the neighbouring populations of countries in the Eastern Baltic region: Estonia, Latvia and Russia. To evaluate the possible contribution of genetic factors to these differences, the frequencies of HLA-DQB1 alleles and relevant DQB1-DQA1 or DQB1-DRB1 haplotypes associated with IDDM risk or protection were analysed among IDDM patients and control subjects from these four populations. An increased frequency of HLA DQB1*0302, DQB1*02-DQA1*05 and DQB1*0302-DRB1*0401 was observed in subjects with IDDM in all studied populations, whereas the prevalence of DQB1*0301 and DQB1*0602 and/or *0603 was decreased among patients. The degree of IDDM risk associated with HLA alleles analysed here did not differ significantly between the populations. Comparisons of the distribution of IDDM-related HLA alleles and haplotypes in the background populations revealed its consonance with IDDM incidence. The combined frequency of high risk genotypes was significantly higher among Finns than in other populations studied. Our data support the hypothesis that variance in the dispersion of HLA alleles is the genetic basis of variation of IDDM incidence observed in the Eastern Baltic region. PMID- 9864038 TI - TAP polymorphism is not associated with ankylosing spondylitis and complications with acute anterior uveitis in HLA-B27-positive Japanese. AB - HLA-B27 is associated with the etiology of ankylosing spondylitis (AS) and acute anterior uveitis (AAU). Transporter associated with antigen processing (TAP) 1 and TAP2 polymorphism influences the range of peptide presented by HLA class I molecules. In this report, contribution of TAP polymorphism to the susceptibility to AS and AAU was studied in HLA-B27-positive Japanese individuals. Patients were classified into three groups: 16 AS patients, 14 AAU patients and 22 patients with both AS and AAU. Twelve HLA-B27-positive healthy individuals were included as a control. TAP polymorphism was detected by PCR-RFLP methods. Significant differences in frequencies of TAP1 alleles were not found between patient groups. None of the TAP2 frequencies showed increased or decreased frequencies compared with HLA-B27-positive healthy controls. In comparison with a random Japanese control, TAP2D allele frequency was significantly increased in the AAU group, but failed to reach a significant level in a group consisting of the AAU-only patients and the patients with both AS and AAU. All of the patient groups were noted to have a significantly increased prevalence of the TAP2H allele as compared to random controls; however, the higher frequency of this allele was detected in HLA-B27 healthy controls as well. These observations suggest a linkage disequilibrium between TAP2D, TAP2H and HLA-B27 in Japanese. PMID- 9864039 TI - High incidence of the Cys 282 Tyr mutation in the HFE gene in the Irish population--implications for haemochromatosis. AB - A simple PCR-SSOP approach based on a single PCR product has been developed to screen the HFE gene for the haemochromatosis-associated mutations Cys 282 Tyr and His 63 Asp. Using this approach the prevalence of these mutations in a cohort (30) of haemochromatosis patients and normal controls (404) was determined. Ninety percent of the haemochromatosis patients were homozygous for the Cys 282 Tyr mutation. In the normal population we found an increased incidence of the Cys 282 Tyr mutation (17.3%; 95% confidence limits 0.136-0.209) which was also reflected in the higher frequency of Cys 282 Tyr homozygotes (1.24%; 95% confidence limits 0.0016-0.0232). Linkage disequilibrium analysis confirmed the association between A*03 and Cys 282 Tyr. However, strong linkage disequilibrium occurred with the HLA-A*03-associated allele HLA-B*14 but not the HLA-A*03 associated allele HLA-B*07. The His 63 Asp was found to be in linkage disequilibrium with HLA-A*29. PMID- 9864041 TI - HLA-B*27 subtyping by PCR-RFLP in Spanish patients with ankylosing spondylitis. AB - We describe the use of restriction analysis on PCR-amplified DNA for detecting all B*27 subtypes except B*2710 and B*2711 (i.e. from B*2701 to B*2709). After detecting B*27 by Sty I, double digestions consisting of Sty I plus another informative enzyme led to subtype assignment. We used mismatched primers to create restriction sites when necessary. The method avoids group-specific amplifications and other laborious optimization procedures. It was successfully tested on a panel of well characterized cell lines covering different B*27 subtypes. Then, we studied a group of 57 ankylosing spondylitis patients and 746 controls from the south of Spain. B*27 showed a very strong association with the disease (OR=211.27, P=10(-7)). B*2702 and B*2705 distribution in controls (20% and 77.1%, respectively) differed from previously reported data in the Spanish population. We unexpectedly found the B*2707 allele in our population (one control). PMID- 9864040 TI - Identification of a novel HLA-B*51 variant: HLA-B*5112. PMID- 9864042 TI - Characterisation of four non-human primate Mhc-DQB1 alleles. PMID- 9864043 TI - Hepatitis C virus NS3/4A protease. AB - Despite an urgent medical need, a broadly effective anti-viral therapy for the treatment of infections with hepatitis C viruses (HCVs) has yet to be developed. One of the approaches to anti-HCV drug discovery is the design and development of specific small molecule drugs to inhibit the proteolytic processing of the HCV polyprotein. This proteolytic processing is catalyzed by a chymotrypsin-like serine protease which is located in the N-terminal region of non-structural protein 3 (NS3). This protease domain forms a tight, non-covalent complex with NS4A, a 54 amino acid activator of NS3 protease. The C-terminal two-thirds of the NS3 protein contain a helicase and a nucleic acid-stimulated nucleoside triphosphatase (NTPase) activities which are probably involved in viral replication. This review will focus on the structure and function of the serine protease activity of NS3/4A and the development of inhibitors of this activity. PMID- 9864044 TI - Recent advances in L-nucleosides: chemistry and biology. PMID- 9864045 TI - Adenovirus endopeptidase and papain are inhibited by the same agents. AB - Adenoviruses encode a cysteine protease (AVP) which carries out highly specific cleavages on at least seven viral proteins and two cellular proteins. Virus infectivity is dependent on this function. The three-dimensional positions of the amino acids involved in catalysis display a striking similarity to those of papain, suggesting a similar catalytic mechanism. This similarity has prompted us to compare the effect of papain inhibitors on the two enzymes. AVP and papain activity was tested on a fluorescent peptide substrate as well as on metabolically labeled adenovirus (Ad2) precursor proteins. Hep2 cells infected with Ad2 were exposed to inhibitors and assayed for, (a) infectious virus, (b) in situ Ad2 protease activity, (c) physical particle production and their polypeptide composition. We found that in both substrate systems AVP was sensitive to the papain inhibitors benzamidoacetonitrile, acetamidoacetonitrile and N-methoxyphenylalanine glycylnitrile, and that the degree of sensitivity was influenced by the substrate. Unlike papain, AVP was relatively insensitive to E64. In ex vivo tests, Hep2 cells infected with Ad2 were exposed to inhibitors and assayed for, (a) infectious virus, (b) in situ Ad2 protease activity, (c) physical particle production and their polypeptide composition. A 4-fold reduction in virus titer was obtained when the inhibitors were added between 17 and 25 h after infection. Processing of precursor proteins was also inhibited yet the production of physical particles was only reduced 2-fold. These experiments show that papain inhibitors are also capable of inhibiting the adenovirus protease both in vitro and ex vivo, thus forging a possible link between structural similarity and functionality. PMID- 9864046 TI - Mycophenolate mofetil strongly potentiates the anti-herpesvirus activity of acyclovir. AB - We demonstrate that the novel immunosuppressive agent mycophenolate mofetil (MMF), that has been approved for use in kidney transplant recipients, strongly potentiates the antiviral activity of acyclovir in murine models for herpesvirus infections. Hairless mice that were infected intracutaneously with herpes simplex virus type 1 were treated systemically with ACV (20 mg/kg per day) and topically with 5% MMF. Combined use of both drugs resulted in an almost complete protection, whereas single use of either compound had virtually no effect. When athymic-nude mice were infected with an ACV-resistant (ACVr)-thymidine kinase deficient (TK-) HSV-2 strain, combined use of systemically administered ACV (100 mg/kg per day) and topically applied MMF (5%) protected 60% of the animals against the infection, whereas all mice treated with either drug alone succumbed. Since transplant recipients under MMF therapy may develop opportunistic herpesvirus infections, requiring treatment with acyclovir (or valaciclovir), our findings have important implications for the treatment of these herpesvirus infections. PMID- 9864047 TI - Therapeutic evaluation of compounds in the SCID-RA papillomavirus model. AB - A previous study by Kreider (Kreider et al., 1979) indicated that rabbit skin, which had been transplanted to immunodeficient nude mice, could be successfully infected with cottontail rabbit papillomavirus (CRPV). We have extended this observation in developing a rodent model for evaluation of compounds for activity against the papillomaviruses. In this model (called the SCID-Ra model), rabbit ear skin is transplanted to the dorsum of SCID mice and allowed to heal for 3 weeks. Infection with CRPV by scarification leads to the growth of warty lesions within 2 3 weeks in >95% of the animals. Topical and/or systemic therapy can be initiated at various times post infection (PI). Weekly lesion scores are recorded and compounds are evaluated for their ability to suppress wart growth when compared to untreated control mice. Ribavirin, which has had a suppressive effect both in the clinic for the treatment of respiratory papillomatosis and on the growth of warts in the rabbit back model, was evaluated and showed significant anti-proliferative activity with oral dosing. Both antiviral and antiproliferative compounds including podophyllin and 5-fluorouracil, which have been used clinically for the treatment of human papillomavirus (HPV) infections, were evaluated in this model. The anti-mitotic compound, Navelbine (vinorelbine tartrate), which is used for the treatment of non-small cell lung carcinoma was evaluated in this system and showed significant inhibition of wart growth with somewhat less topical cytotoxicity when compared to podophyllotoxin. PMID- 9864048 TI - Comparison of antiviral compounds against human herpesvirus 6 and 7. AB - Four classes of antiviral compounds were evaluated for inhibitory activity against two variants of human herpesvirus 6 (HHV-6A and -6B) and human herpesvirus 7 (HHV-7). These included: (1) a pyrophosphate analog, phosphonoformic acid (PFA); (2) beta-guanine analogs, 9-(2 hydroxyethoxymethyl)guanine (acyclovir or ACV), 9-[(1,3-dihydroxy-2 propoxy)methyl]guanine (ganciclovir or GCV) and 9-(4-hydroxy-3-hydroxy-3 hydroxymethylbutylyl)guanine (penciclovir or PCV); (3) acyclic nucleoside phosphonates, (S)-1-[(3-hydroxy-2-phosphonylmethoxy)propyl]cytosine [cidofovir or (S)-HPMPC] and its cyclic derivative (S)-cyclic-HPMPC (cHPMPC), 9-[[2-hydroxy-1 phosphonomethoxy)ethoxy]methyl]guanine (HPMEMG) and 9-[(2 phosphonylmethoxy)ethyl]-2,6-diaminopurine (PMEDAP), and the seven other related compounds; and (4) a series of benzimidazole ribonucleosides, including 2-bromo 5,6-dichloro-1-(beta-D-ribofuranosyl)benzimidazole (BDCRB). End-point inhibitory concentration (EPC) and 50% effective inhibitory concentration (EC50) values were determined by a dot-blot antigen detection method in cord blood mononuclear cells infected with HHV-6A, HHV-6B or HHV-7 at a multiplicity of infection of 0.004 CCID50/cell. (S)-HPMPC and cHPMPC had an EC50 value of approximately 0.3 microg/ml for HHV-6A, 1.2 microg/ml for HHV-6B and 3.0 microg/ml for HHV-7. These compounds were the most active of those tested against each virus. The EC50 value of GCV for HHV-6A was 0.65 microg/ml, 1.33 microg/ml for HHV-6B, and >7 microg/ml for HHV-7. The EC50 values of ACV and PCV were approximately 6-8 microg/ml for HHV-6A, 16-24 microg/ml for HHV-6B and 121-128 microg/ml for HHV-7. These drugs were the least active. The sensitivity of HHV-7 to the guanine analogs was different from HHV-6, suggesting a difference in selectivity of specific viral enzymes. PMID- 9864050 TI - Genetic analysis of a ganciclovir-resistant human cytomegalovirus mutant. AB - We isolated a ganciclovir (GCV)-resistant human cytomegalovirus (HCMV) from a laboratory strain, AD169, and analysed the mutant. Attempts were also made to identify directly the mutated gene. The 50% inhibitory concentration (IC50) of GCV for the mutant strain was five times higher than that of the wild-type strain. The mutant strain showed similar sensitivity to phosphonoacetic acid and cidofovir as the wild-type strain. These data suggest mutation in the UL97 gene encoding for the phosphotransferase that phosphorylates GCV. Molecular analysis of the mutant strain revealed that a single base substitution of adenine by cytosine occurred at the 1796 nucleotide position of the UL97 gene region, resulting in the substitution of lysine by threonine at codon 599 in the UL97 gene product. Marker transfer experiment confirmed that this mutation conferred HCMV resistance to GCV. The mutation at codon 599 was easily identified by means of RsaI digestion of the selected PCR product. PMID- 9864049 TI - The anti-herpes simplex virus activity of n-docosanol includes inhibition of the viral entry process. AB - n-Docosanol-treated cells resist infection by a variety of lipid-enveloped viruses including the herpesviruses. Previous studies of the mechanism of action demonstrated that n-docosanol inhibits an event prior to the expression of intermediate early gene products but subsequent to HSV attachment. The studies reported here indicate that n-docosanol inhibits fusion of the HSV envelope with the plasma membrane. Evidence suggests that antiviral activity requires a time dependent metabolic conversion of the compound. Cellular resistance to infection declines after removal of the drug with a t1/2 of approximately 3 h. Reduced expression of viral genes in n-docosanol-treated cells was confirmed by a 70% reduction in expression of a reporter gene regulated by a constitutive promoter inserted into the viral genome. Inhibited release in treated cells of virion associated regulatory proteins--an immediate post entry event--was indicated by a 75% reduction in the expression of beta-galactosidase in target cells carrying a stably transfected lacZ gene under control of an HSV immediate--early promoter. Finally, the fusion-dependent dequenching of a lipophilic fluorescent probe, octadecyl rhodamine B chloride, inserted into the HSV envelope was significantly inhibited in treated cells. Inhibition of fusion between the plasma membrane and the HSV envelope, and the subsequent lack of replicative events, may be the predominant mechanism for the anti-HSV activity of n-docosanol. PMID- 9864051 TI - Solid phase ELISA for determination of the virus dose dependent sensitivity of human cytomegalovirus to antiviral drugs in vitro. AB - The main problems in determining the true in vivo susceptibility of human cytomegalovirus (CMV) to antiviral compounds are the influence of the size of the viral inoculum, the variation in the replication capacity of different CMV strains and the subjective evaluation of the inhibition of viral growth in the plaque assay. In this study, a specific assay was developed which reproducibly determines the sensitivity of primary isolates of CMV. The assay includes simultaneous virus titration and determination of the antiviral sensitivity. When individual virus doses were evaluated, the IC50 was generally dependent on the virus dose, except for resistant isolates, where the IC50 did not change irrespective of the dose of virus. The novel method of IC50 calculation takes into account all values derived from the linear part of the inhibition curve. This may better reflect the in vivo conditions, where the antiviral drug encounters different amounts of virus in different organs. Two human fibroblast derived cell lines showed similar results. PMID- 9864052 TI - Immunodominant peptide epitopes of allergen, Asp f 1 from the fungus Aspergillus fumigatus. AB - Aspergillus fumigatus ribotoxin Asp f 1 is a major allergen with IgE binding activity to serum of a majority of patients with allergic bronchopulmonary aspergillosis (ABPA). The IgE binding epitopes or the T-cell stimulatory peptides of this molecule have not been studied. In the present investigation, we have synthesized linear decapeptides spanning the whole molecule of Asp f 1 and analyzed their IgE binding properties. We have also synthesized peptides based on their possible T-cell stimulatory properties and studied the stimulation of peripheral blood mononuclear cells from ABPA patients and normal controls. Several peptides demonstrated distinct IgE antibody binding response against sera from ABPA patients and proliferative response against peripheral blood mononuclear cells from the patients. From the results, it can be concluded that the carboxy-terminal region of Asp f 1 representing amino acid residues 115-149 involved in both humoral and cell mediated immunoresponses in ABPA. PMID- 9864053 TI - Peptides related to FKBP 39-45 loop possess immunostimulative potency. AB - Taking into account the sequential homology existing between thymopoietin II, the DNA-binding domain of p53 protein and FKBP (FK-506 binding protein), a series of fragments of human and bovine FKBP containing a fragment Ser39-Pro45 were synthesized. In the humoral in vitro test all the peptides act as stimulators. Whereas in the in vivo test peptides derived from bovine FKBP show an immunostimulative and those from human FKBP an immunosuppressive activity. However, after blocking the Asp residue by a Bzl group the peptide V appears to be an immunostimulator. The data obtained suggest that these peptides can influence the immune system by blocking the FKBP receptor. PMID- 9864054 TI - Purification and primary structure of pituitary adenylate cyclase activating polypeptide (PACAP) from the brain of an elasmobranch, stingray, Dasyatis akajei. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) was isolated from ovine hypothalami and found to exist as two amidated forms with 38 (PACAP 38) and 27 (PACAP 27) residues. The amino acid sequences of PACAPs isolated from the vertebrates, such as a bird, a frog and teleost fish, appear to be well conserved. In the present study, we attempted to isolate PACAP from the brain of an elasmobranch fish, Dasyatis akajei (stingray), which belongs to the Chondrichthyes (cartilaginous fish), by extraction of the acetone-dried powder with acetic acid, followed by successive high-performance liquid chromatography (HPLC) on a gel-filtration, a cation-exchange and two reverse-phase columns. Purification was monitored by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and Western blotting analysis using an anti-PACAP 27 serum. The PACAP thus obtained consisted of 44 residues. The amino acid sequence of the comparable portion of its N-terminal 38 residues showed 92%, 89%, 89%, and 82% identity with those of mammalian, chicken, frog and teleost PACAPs with 38 residues, respectively. The extra six C-terminal residues of the stingray resembled those of tetrapod and teleost PACAP precursors which were deduced from the respective cDNAs. These results indicate that PACAP, which has an amino acid sequence showing high similarity with those of tetrapod and teleost PACAPs, is present in the elasmobranch brain. PMID- 9864056 TI - Inhibitory effects of peptide YY on basal and VIP-stimulated short-circuit current in the rat jejunum: influence of technical conditions on observed results. AB - The interaction of PYY and VIP was studied in stripped and intact rat jejunum preparations mounted in Ussing chambers. PYY decreased basal Isc in intact as well as in stripped jejunum. Stripping was necessary to evidence a stimulation of basal Isc by VIP. When PYY and VIP were administered at the same time in the serosal bath, their effects seemed additive; VIP stimulation took over when VIP was present in ten times larger amounts than PYY, while PYY inhibition predominated at isomolar concentrations (10(-7) M) of both peptides. However, when PYY was administered three to six minutes before isomolar amounts of VIP, the VIP stimulation developed without being notably hampered. At this time, however, the amount of radioimmunoassayable PYY in the serosal compartment represented still 60% of the added amount. In conclusion, the experimental conditions can significantly change the results: stripping the longitudinal muscle/myenteric plexus impairs the effect of PYY and VIP in a different fashion, while the timing and order of administration of the peptides may change the apparent interaction between VIP stimulation and PYY inhibition. PMID- 9864055 TI - The augmentation of pituitary adenylate cyclase-activating polypeptide (PACAP) in streptozotocin-induced diabetic rats. AB - Pituitary adenylate cyclase activating polypeptide (PACAP), which was isolated from ovine hypothalamic extract, has been shown to have a physiological role in the regulation of insulin or islet functions. In streptozotocin (STZ)-induced diabetic rats, we examined the content of PACAP immunoreactivity and gene expression of three specific receptors. Four weeks after administration of STZ (50 mg/kg), plasma glucose levels increased 3.3-fold, and plasma insulin levels decreased to one-tenth as compared with the control. The content of PACAP immunoreactivity in the pancreas potently increased by 30%, but the content of vasoactive intestinal polypeptide (VIP) immunoreactivity was not changed. In the other tissues, the content of PACAP immunoreactivity did not significantly change except in the hypothalamus, which showed a 10% increment. In the expression level of PACAP/VIP receptors, semi-quantitative RT-PCR analysis revealed that VIP1/PACAP receptor mRNA significantly increased as compared with the other two types of receptors in the pancreas of STZ-induced diabetic rats. These findings suggest that PACAP and VIP1/PACAP receptor might be involved in the pathophysiology of diabetes mellitus. PMID- 9864057 TI - Human adrenocortical NCI-H295 cells express VIP receptors. Steroidogenic effect of vasoactive intestinal peptide (VIP). AB - VIP receptors are frequently overexpressed by various endocrine tumors. In this study the expression of VIP receptors in the human adrenocortical carcinoma cell line NCI-H295 and their involvement in the regulation of steroidogenesis was investigated. NCI-H295 cells express VIP1 and VIP2 receptors as demonstrated by RT-PCR, whereas they do not express VIP itself. The receptors are functionally coupled to steroidogenesis since VIP (10(-9) M to 10(-6) M) exerted a dose dependent stimulatory effect on the release of aldosterone, cortisol, and DHEA. VIP increased ACTH-stimulated releases of aldosterone and cortisol. The proliferation rate of NCI-H295 cells was not affected by VIP. These data show that NCI-H295 cells express both forms of the VIP receptor and that VIP is involved in an ACTH-independent regulation of steroidogenesis in the adrenal tumor cells. PMID- 9864058 TI - Induction of apoptosis by a short-chain neuropeptide analog in small cell lung cancer. AB - Small cell lung cancer (SCLC) cells express a variety of neuropeptides which act as autocrine growth factors. Although several neuropeptide analogs have been reported to antagonize SCLC proliferation, the development of these compounds has been limited by their low potency and the cytostatic nature of their effects. In the present study we evaluated the cytotoxic activity of four short-chain substance P analogs (NY3460, NY3238[-pHOPA], NY3238[Phe1], NY3238[Lys5]) against a panel of five SCLC cell lines. NY3460 was the most potent compound in all five SCLC cell lines (IC50 = 2.8-3.7 microM) as assessed by a MTT growth inhibitory assay. NY3238[Phe1] was also relatively active in all cell lines (IC50 = 3.5-11.2 microM), while NY3238[Lys5] and NY3238[-pHOPA] were substantially less active. NY3460 was the only agent to induce an increase in the percentage of cells with subdiploid DNA content suggestive of apoptosis by flow cytometric DNA content analysis. The induction of apoptosis was confirmed by fluorescent microscopy in NCI-H69, NCI-H82, NCI-H446, and NCI-H510 cells after exposure to 5.0 microM NY3460 for 48 h. These findings suggest that NY3460 is a relatively potent cytotoxic inhibitor of SCLC growth, and that short-chain neuropeptide analogs deserve further evaluation as anti-SCLC agents. PMID- 9864059 TI - Binding of enterostatin to the human neuroepithelioma cell line SK-N-MC. AB - SK-N-MC cells were found to possess binding sites for enterostatin, a peptide with central effects on appetite and sympathetic activation of brown adipose tissue during high-fat feeding. Scatchard analyses of the binding indicated one high-affinity binding (Kd = 0.5-1.5 nM) and one low-affinity binding (Kd = 15-30 nM) for 3H-enterostatin (APGPR). 125I-YGGAPGPR showed similar binding parameters as for the low affinity binding of 3H-APGPR. Met-enkephalin and beta3 casomorphin1-5 were found to displace the binding of 3H-APGPR to the SK-N-MC cells. Affinity purification of solubilized cells revealed an APGPR-binding protein estimated to 53 kDa which may represent a distinct enterostatin receptor. Cross-linking of 125I-YGGAPGPR to intact cells labeled one major protein with the same molecular size. There was no binding of enterostatin to four other human neuroblastoma/neuroepithelioma cell lines, named IMR-92, LAN#5, NB-1 #14 and SH5 SY. PMID- 9864060 TI - Amylin reduces food intake more potently than calcitonin gene-related peptide (CGRP) when injected into the lateral brain ventricle in rats. AB - Amylin and the structurally and functionally related peptide calcitonin gene related peptide (CGRP) have been shown to reduce food intake in rats. The aim of the present study was to compare the anorectic potency of both peptides over a wide dose range when administered into the lateral brain ventricle (ICV). Furthermore, we also tested the influence of a lesion in the area postrema/nucleus of the solitary tract (AP/NTS) region on the anorectic effects of amylin and CGRP after ICV administration because AP/NTS lesion has been shown to reduce the anorectic effects of both peptides when injected intraperitoneally (IP). Amylin [1-510 pmol/rat (0.004-2 microg/rat) ICV] and CGRP [1-131 pmol/rat (0.004-0.5 microg/rat) ICV] dose-dependently reduced food intake in food-deprived rats. At a dose of 26 pmol/rat (0.1 microg/rat), amylin almost completely suppressed food intake for 1 h after injection. Amylin [EC50 = 2 pmol/rat (0.007 microg/rat)] was markedly more potent than CGRP [57 pmol/rat (0.215 microg/rat)] with regard to its anorectic effect. A lesion in the AP/NTS region did not influence the anorectic effects of amylin and CGRP after administration into the lateral ventricle. It is concluded that amylin is more potent than CGRP in reducing food intake after administration into the lateral brain ventricle. Receptors in the forebrain may mediate the anorectic effects of both peptides when administered via this route. PMID- 9864061 TI - Soybean agglutinin stimulated cholecystokinin release from cultured rabbit jejunal cells requires calcium influx via L-type calcium channels. AB - We have studied the mechanism of soybean agglutinin (SBA) mediated cholecystokinin (CCK) release in enriched cultured cholecystokinin-secreting cells. 12-O-Tetradecanoylphorbol-13-acetate 1 mM significantly stimulated release of CCK-like-immunoreactivity (CCK-LI) by 55%+/-17% (p < 0.05), which was blocked by the protein kinase C inhibitor staurosporine 100 nM. Forskolin 10 mM stimulated CCK-LI by 82%+/-12% (p < 0.05) and this was inhibited by somatostatin 1 nM. 1-Phenylalanine 20 mM and Bay K 8644 1 mM stimulated CCK-LI by 69%+/-22% and 60%+/-19% respectively (p < 0.05), these responses were completely abolished by the L-type calcium channel antagonist verapamil 10 mM. SBA 10 and 100 microg/ml stimulated CCK-LI by 65%+/-22% and 74%+/-24% respectively (p < 0.05). The effect of SBA was inhibited by verapamil and N-acetylgalactosamine. We conclude that SBA stimulates CCK-LI through calcium flux via L-type calcium channels. PMID- 9864062 TI - Hyperphagia induced by hypoglycemia in rats is independent of leptin and hypothalamic neuropeptide Y (NPY). AB - Hypoglycemia causes hyperphagia and weight gain, through unknown peripheral and central signals. We investigated the effect of hypoglycemia on NPY and leptin expression and the ability of leptin to inhibit hypoglycemia-induced hyperphagia. Acute hypoglycemia (60 U/kg SC insulin; n = 8) increased food intake (p < 0.01) compared with controls (n = 8). Insulin- and leptin-treated rats (300 microg/kg IP leptin; n = 8) had reduced hyperphagia (p < 0.05 vs. controls; p < 0.05 vs. insulin alone) and a 15% fall in NPY mRNA levels compared with controls (p < 0.01). Chronic hypoglycemia, (20-60 U/kg/day insulin; n = 8) increased food intake compared with vehicle-treated controls (p < 0.01). Leptin and insulin administration (300 microg/kg/day IP leptin; n = 8) reduced hyperphagia (p < 0.01 vs. controls, p < 0.05 vs. insulin alone), and NPY mRNA fell by 18% vs. controls (p < 0.01). We conclude that hypoglycemia-induced hyperphagia is not mediated by either a fall in leptin or an increase in hypothalamic NPY mRNA. Leptin can inhibit feeding in hyperphagic hypoglycemic rats, and this may partly be attributable to its inhibition of the NPY neurons. PMID- 9864063 TI - Application of taste reactivity to study the mechanism of alcohol intake inhibition by the tachykinin aminosenktide. AB - Tachykinin NK-3 receptor agonists reduce alcohol intake in alcohol-preferring rats; the nucleus basalis magnocellularis (NBM) is highly sensitive to their effect. Tachykinins and their receptors are widely distributed in gustatory pathways and NK-3 receptor agonists have been reported to modify taste reactivity to salt solutions in rats. The present study evaluated whether the TK NK-3 receptor agonist aminosenktide (NH2-SENK) influences taste reactivity to ethanol solutions. Genetically selected Marchigian Sardinian alcohol-preferring (msP) rats were employed. In response to the intraoral infusion (0.8 ml in 1 min) of 10, 20, 40, or even 60% ethanol solution, ethanol-naive rats showed a large number of ingestive reactions and a much lower number of aversive reactions. Two min before the intraoral infusion of 10 or 40% ethanol, NH2-SENK was injected either into the lateral ventricle (LV) or into the NBM. Doses of NH2-SENK that markedly reduce alcohol intake, 125 ng/rat into the LV or 5 ng/site into the NBM, did not modify the ingestive reactions and, in some instances, reduced the aversive reactions to ethanol solutions in ethanol-naive rats. Injections of 125 ng/rat into the LV failed to modify taste reactions in ethanol-experienced rats. The present results show that msP rats have an innate hedonic evaluation of ethanol solutions, even of high concentration. Moreover, they indicate that reduction of ethanol intake induced by TK NK-3 receptor agonists in alcohol preferring rats does not depend on influences on gustatory processes. PMID- 9864064 TI - Somatostatin administration alters taste preferences in the rat. AB - The effects of long-term and relatively constant concentrations of somatostatin (SRIF) were tested in rats to investigate its influence on taste preferences, fluid intake, and taste bud topography of the tongue. In SRIF-treated rats, intake of 3 x 10(-2) M NaCl and 6 x 10(-2) M sucrose solution decreased, whereas that of 3 x 10(-5) M quinine-HCl and 10(-3) M HCl (pH = 3) solutions increased significantly; distilled water intake and total fluid consumption remained almost unchanged. The light/dark cycle of consumption of NaCl solution was modified in a dose-dependent manner, but that of dwater, total fluids, and other solutions was not. Histologic exam by light microscopy showed that taste bud distribution on the tongue appeared to be altered, but the results only approached statistical significance. Given that it is possible to remove virtually all taste buds in the oral cavity and have only modest effects on preferences and aversion, it seems very unlikely that the small changes in taste bud distribution are related to the intake of sapid solutions. Since SRIF treatment influenced all sapid stimuli, it seems more likely that the changes are based on a blunted responsiveness in the gustatory system as a whole. PMID- 9864065 TI - Somatostatin decreases somatostatin messenger ribonucleic acid levels in the rat periventricular nucleus. AB - Growth hormone (GH) secretion from the pituitary is known to be under the dual control of GH-releasing factor (GRF) and somatostatin (SRIF). Hypothalamic SRIF, the major inhibitor of pituitary growth hormone secretion, inhibits its own release by a negative ultrashort-loop feedback mechanism. However, it is not known whether this negative regulation is mediated by inhibition of SRIF mRNA production. GRF may also inhibit its own release, thereby modifying pituitary GH secretion, possibly through an ultrashort-loop feedback mechanism. Thus, SRIF production and GRF release are both regulated by SRIF. Periventricular nucleus (PeN) and mediobasal hypothalamus (MBH) from adult male rats were incubated for 6 h in Waymouth's medium with either SRIF or the SRIF agonist analog RC 160 (10(-9) to 10(-6) M). Levels of SRIF mRNA were determined by an S1 nuclease protection assay using a 32[P]-labeled rat SRIF riboprobe. SRIF (10(-7) M) and RC 160 (10( 8), 10(-7) M) significantly (p< or =0.01) decreased SRIF mRNA levels in the PeN. The levels of SRIF mRNA in the MBH were not modified by either SRIF or RC 160. SRIF (10(-7) and 10(-6) M) significantly (p < or = 0.01 and p < or = 0.001, respectively) inhibited the release of GRF at 30 min in the MBH. Likewise, the release of GRF was slightly decreased by 10(-7) M RC 160, and significantly inhibited by 10(-6) M (p < or = 0.001) at 30 min. At 6 h, the levels of GRF were significantly reduced by 10(-7) M SRIF (p < or = 0.05) and by RC 160 (10(-7), 10( 6) M; p < or = 0.001 and p < or = 0.05, respectively). In contrast with these results, the SRIF analog was unable to alter SRIF release at 30 min. At 6 h incubation, RC 160 (10(-7) M) significantly (p < or = 0.001) reduced SRIF release from MBH fragments. These results demonstrate that SRIF and a SRIF analog decrease SRIF mRNA levels in the PeN and inhibit the release of SRIF from the nerve terminals of the MBH. Thus, SRIF appears to regulate its own gene expression by negative ultrashort-loop feedback. Therefore, when SRIF is secreted from these neurons in response to GRF, it down-regulates the preceding stimulatory input as well as its own secretion. PMID- 9864066 TI - Autoradiographic evidence that zona glomerulosa and capsular vessels of the human adrenal cortex are provided with different subtypes of adrenomedullin receptors. AB - Frozen sections of normal adrenal glands, obtained from patients undergoing unilateral nephrectomy for kidney cancer, were labeled in vitro with human [125I]ADM(1-52). Autoradiography and quantitative densitometry showed the presence of abundant ADM(1-52) binding sites in both zona glomerulosa (ZG) and capsular vessels, which were displaced with about the same efficiency by cold ADM(1-52) and rat ADM(1-50). The selective calcitonin gene-related peptide type 1 (CGRPI) ligand CGRP(8-37) eliminated, although less efficiently than ADMs, [125I]ADM(1-52) binding in the ZG, but not in the capsular vessels. These findings suggest the existence of different receptor subtypes for ADM in the human adrenal cortex. The CGRP(8-37)-sensitive receptors located in the ZG may mediate the well-known inhibitory effect of ADM on aldosterone secretion, while the CGRP(8-37)-insensitive receptors present in the capsular vessel may be involved in the ADM-induced rise in adrenal blood flow. PMID- 9864067 TI - Endopeptidase 24.11 inhibition does not modify uterotonic effects of endothelins in rat uterus. AB - We investigated effects of the endopeptidase 24.11 inhibitor, SCH 39370, on uterotonic effects of endothelins (ETs) and sarafotoxin S6b. Responses of uteri from non-pregnant rats were inhibited by the ETA receptor antagonist, BQ123 (1 microM) but not the ETB receptor antagonist, BQ 788 (1 microM). ET-1, sarafotoxin S6b and ET-2 were more potent than ET-3 in tissues from non-pregnant and pregnant rats. SCH 39370 (10 microM) did not affect uterotonic responses to these peptides in either group, but inhibited those of big ET-1 in non-pregnant rat tissues, indicating inhibition of conversion of big ET-1 to ET-1. These data indicate that endopeptidase 24.11 does not inactivate the endothelin peptides in the rat uterus. PMID- 9864068 TI - Nitric oxide release mediates vasodilator responses to endomorphin 1 but not nociceptin/OFQ in the hindquarters vascular bed of the rat. AB - We have recently shown that endomorphin 1, an endogenous ligand for the mu-opioid receptor, and nociceptin (Orphanin FQ; OFQ), an endogenous ligand for the ORL1 receptor, have substantial vasodilator activity in the hindquarters vascular bed of the rat. In the present study, the role of nitric oxide, vasodilator prostaglandins, and the opening of K+ ATP channels in mediating vasodilator responses to endomorphin 1, PL017, and DAMGO was investigated in the regional vascular bed in the rat. Under constant-flow conditions, injections of the mu selective agonists endomorphin 1, PL017 ([N-MePhe3,D-Pro4]-morphiceptin), and DAMGO, and the ORL1 receptor agonist nociceptin/ OFQ produced dose-dependent decreases in hindquarters perfusion pressure. Vasodilator responses to endomorphin 1, PL017, and DAMGO, and the endothelium-dependent vasodilators acetylcholine and adrenomedullin were attenuated by the nitric oxide synthase inhibitor L-NAME (50 mg/kg IV) at a time when vasodilator responses to nociceptin/OFQ were not altered. Vasodilator responses to isoproterenol and prostaglandin E1, agents known to increase cAMP levels, and the nitric oxide donor DEA/NO were not altered by the nitric oxide synthase inhibitor. Responses to endomorphin 1, PL017, DAMGO, and nociceptin/OFQ were not altered by sodium meclofenamate at a time when vasodilator responses to arachidonic acid were reduced significantly or after administration of U-37883A at a time when vasodilator responses to levcromakalim were reduced significantly. The results of these studies indicate that responses to endomorphin 1, PL017, and DAMGO are mediated in large part by the release of nitric oxide, while responses to nociceptin/OFQ are mediated by an L-NAME-insensitive mechanism. Moreover, these results demonstrate that responses to these peptides are not mediated by the release of vasodilator prostaglandins or the opening of K+ATP channels the hindquarters vascular bed. PMID- 9864069 TI - Opiate tolerance and dependence: receptors, G-proteins, and antiopiates. AB - Despite the existence of a large body of information on the subject, the mechanisms of opiate tolerance and dependence are not yet fully understood. Although the traditional mechanisms of receptor down-regulation and desensitization seem to play a role, they cannot entirely explain the phenomena of tolerance and dependence. Therefore, other mechanisms, such as the presence of antiopiate systems and the coupling of opiate receptors to alternative G proteins, should be considered. A further complication of studies of opiate tolerance and dependence is the multiplicity of endogenous opiate receptors and peptides. This review will focus on the endogenous opioid system--peptides, receptors, and coupling of receptors to intracellular signaling via G-proteins- in the context of their roles in tolerance and dependence. Opioid peptides include the recently discovered endomorphins and those encoded by three known genes--pro-opiomelanocortin, pro-enkephalin, and pro-dynorphin. They bind to three types of receptors--mu, delta, and kappa. Each of the receptor types is further divided into multiple subtypes. These receptors are widely known to be coupled to G-proteins of the Gi and Go subtypes, but an increasing body of results suggests coupling to other G-proteins, such as Gs. The coupling of opiate receptors to Gs, in particular, has implications for tolerance and dependence. Alterations at the receptor and transduction level have been the focus of many studies of opiate tolerance and dependence. In these studies, both receptor down regulation and desensitization have been demonstrated in vivo and in vitro. Receptor down-regulation has been more easily observed in vitro, especially in response to morphine, a phenomenon which suggests that some factor which is missing in vitro prevents receptors from down-regulating in vivo and may play a critical role in tolerance and dependence. We suggest that antiopiate peptides may operate in vivo in this capacity, and we outline the evidence for the antiopiate properties of three peptides: neuropeptide FF, orphanin FQ/nociceptin, and Tyr-W-MIF-1. In addition, we provide new results suggesting that Tyr-W-MIF-1 may act as an antiopiate at the cellular level by inhibiting basal G-protein activation, in contrast to the activation of G-proteins by opiate agonists. PMID- 9864070 TI - Oleoyl-estrone does not alter hypothalamic neuropeptide Y in Zucker lean and obese rats. AB - Female Zucker lean and obese rats were treated for 14 days with 3.5 micromol/kg oleoyl-estrone (OE) in liposomes (Merlin-2). After 0, 3, 6, 10, and 14 days of treatment, the rats were killed and hypothalamic nuclei (lateral preoptic, median preoptic, paraventricular, ventromedial and arcuate) were used for neuropeptide Y (NPY) radioimmunoassay. In 14 days, OE decreased food intake by 26% in lean and 38% in obese rats and energy expenditure by 6% in lean and 47% in obese rats; the body weight gap between controls and treated rats becoming -17.8% of initial b.wt. in the lean and -13.6% in the obese rats. Obese rats showed higher NPY levels in all the nuclei than the lean rats. Despite a negative energy balance and decreased food intake, there were practically no changes in NPY with OE treatment. The results indicate that oleoyl-estrone does not act through NPY in its control of either food intake or thermogenesis in lean and genetically obese rats. PMID- 9864071 TI - Sex differences and in vitro effects of steroids on serum aminopeptidase activities. AB - We studied the possible existence of physiological sex differences in serum aminopeptidase activities in mice, by evaluating the effect of gonadectomy and the in vitro response to the presence in the medium of cholesterol or steroid hormones. Alanyl- and glutamyl-aminopeptidase activities were measured in sera from male, female, orchiectomized and ovariectomized mice, incubated with substrate solutions, and compared with the same groups of serum incubated with substrate solutions including cholesterol, 17-beta-estradiol, testosterone, progesterone or hydrocortisone. Our results demonstrated highly significant sex differences, and an influence of cholesterol and steroid hormones on aminopeptidase activity. Depending on the nature of the aminopeptidase, these enzymes responded in different ways to the presence of these substances and also responded differently to gonadectomy. For alanyl-aminopeptidase activity, but not for glutamyl-aminopeptidase activity, there was a clear difference in response between males and females to incubation of the serum with steroid hormones. PMID- 9864072 TI - Potassium, antipsychotic agents, arrhythmias, and sudden death. PMID- 9864073 TI - Atypical features and treatment response in the National Institute of Mental Health Treatment of Depression Collaborative Research Program. AB - Because depression with atypical features is poorly responsive to imipramine, treatment trials including a tricyclic antidepressant arm should assess depressive subtype. Sotsky and Simmens had previously reanalyzed data from the National Institute of Mental Health Treatment of Depression Collaborative Research Program (TDCRP) providing independent confirmation that imipramine is ineffective for patients with atypical features. The TDCRP was a 16-week study in which 239 outpatients with major depression were randomly assigned to cognitive behavior therapy (CBT), interpersonal psychotherapy (IPT), imipraminecase management (IMI-CM), or pill placebo-case management (Pbo-CM). We used Sotsky and Simmens' algorithm to investigate the effect of diagnostic subtype on all four treatments. Hierarchical multiple regression analyses demonstrated IMI-CM benefit relative to Pbo-CM in patients without but not in those with atypical features. These analyses did not demonstrate differential psychotherapy efficacy between depressive subtypes. In conclusion, subsequent analyses of the TDCRP study demonstrated the need to identify the subgroup of depressed patients who have atypical features. Failure to identify this subtype underestimates imipramine's benefit in the appropriately treated subgroup. Comparisons of other treatments with imipramine may be misleading if they do not account for diagnostic subtype. PMID- 9864075 TI - Faster onset of action of fluvoxamine in combination with pindolol in the treatment of delusional depression: a controlled study. AB - This double-blind, controlled study was undertaken to investigate whether the addition of pindolol could improve the therapeutic response to fluvoxamine of depressed patients with psychotic features. After a 1-week placebo run-in period, 72 patients received fluvoxamine 300 mg/day in combination with placebo or pindolol 7.5 mg/day. At study completion, 28 (80%) of 35 patients treated with fluvoxamine plus placebo and 29 (80.5%) of 36 patients treated with fluvoxamine plus pindolol were categorized as responders (reduction of their score on the 21 item Hamilton Rating Scale for Depression to 8 or less and on the Dimension for the Delusional Experience Rating Scale to 0). In the third and fourth weeks, the response rates were significantly superior in the fluvoxamine plus pindolol group (p = 0.0001, p = 0.023, respectively). Treatment response seemed to be unrelated to the demographic and the clinical characteristics recorded. No significant difference was found comparing plasma levels of fluvoxamine between groups, thus excluding a pharmacokinetic interaction. Other than mild nausea and sedation in a few patients, treatments were well tolerated. No medically significant adverse events occurred. Depressed patients with psychotic features who were administered pindolol experienced a more rapid improvement during fluvoxamine treatment. Thus, the combination of fluvoxamine with pindolol may be a useful pharmacologic strategy in the treatment of this disorder. A rapid clinical response in such patients is of relevance in clinical practice as well as in economic fields, given the direct and indirect costs of depression. PMID- 9864074 TI - Efficacy and safety of fluoxetine in treating bipolar II major depressive episode. AB - As many as 45% of patients with major depressive episode also meet DSM-IV criteria for bipolar II (BP II) disorder. Although some clinicians advocate using a mood stabilizer in treating BP II depression, antidepressant monotherapy has been less well studied in this disorder. As part of a prospective, placebo controlled, relapse-prevention study in 839 patients, the efficacy and safety of short- and long-term fluoxetine treatment in patients with BP II major depression compared with patients with unipolar (UP) major depression was retrospectively examined. Eighty-nine BP II patients (mean age, 41+/-11 years) were compared with 89 age- and gender-matched UP patients and with 661 unmatched UP patients (mean age, 39+/-11 years). All received short-term fluoxetine therapy at 20 mg daily for up to 12 weeks. Complete remission was defined as a final Hamilton Rating Scale for Depression score < or = 7 by week 9 that was then maintained for 3 additional weeks. Remitted patients were then randomly assigned to receive double blind treatment with one of the following: (1) fluoxetine 20 mg daily for 52 weeks; (2) fluoxetine for 38 weeks, then placebo for 14 weeks; (3) fluoxetine for 14 weeks, then placebo for 38 weeks; or (4) placebo for 52 weeks. Antidepressant efficacy was similar in BP and UP patients during short-term therapy. Discontinuation for lack of efficacy was lower in BP II (5%) than in UP (12%) patients (p = not significant [NS]), whereas dropouts for adverse events were similar in BP II (11%) and UP (9%) patients. During long-term relapse-prevention therapy, relapse rates were similar in BP II and UP patients (p = NS). During short-term fluoxetine therapy, three BP II (3.8%) versus no matched UP (p = NS) and 0.3% unmatched UP (p = 0.01) patients had a "manic switch." During long-term fluoxetine therapy, one (2%) BP II and three (1%) unmatched UP patients (one taking placebo) had a manic switch (p = NS). In conclusion, fluoxetine may be a safe and effective antidepressant monotherapy for the short-term treatment of BP II depression with a relatively low manic switch rate. Fluoxetine may also be effective in relapse-prevention therapy in patients with BP II disorder. PMID- 9864076 TI - Involvement of serotonin and dopamine in the mechanism of action of novel antidepressant drugs: a review. AB - Several hypotheses regarding the physiopathology of major depression exist. Attention has been focused on cerebral monoaminergic systems, the dysfunction of which is thought to underlie various aspects of depressive symptomatology. There is extensive literature describing the involvement of serotonergic and dopaminergic systems in the mechanism of action of antidepressant drugs. However, a unitary analysis of the data in terms of interaction between different monoaminergic systems is still lacking. In this article, studies reporting the biochemical, behavioral, and clinical effects of tricyclic antidepressants (TCAs), monoamine oxidase inhibitors (MAOIs), selective serotonin reuptake inhibitors (SSRIs), selective blockers of presynaptic dopamine (DA) receptors, and antagonists of serotonin-2 (5-hydroxytryptamine-2 [5-HT2]) receptors were reviewed. Analysis of the current literature indicates that long-term treatment with antidepressants causes adaptive changes of the serotonergic and dopaminergic systems. In particular, long-term administration of TCAs enhances the responsiveness of postsynaptic serotonin receptors to iontophoretically applied serotonin and potentiates the behavioral responses to both direct and indirect dopaminergic agonists. Repeated administration of SSRIs and MAOIs increases serotonergic transmission by desensitizing the inhibitory 5-HT1A somatodendritic and terminal 5-HT1B/1D autoreceptors. Selective blockers of DA autoreceptors exert their antidepressant effect by enhancing DA release. A similar mechanism of action could be hypothesized for 5-HT2 receptor antagonists. There is general agreement that the clinical effect of antidepressant drugs, which becomes evident only after long-term treatment, is caused by their ability to induce adaptive changes of the monoaminergic systems. Increases in both serotonergic and dopaminergic function have been consistently found after long-term treatment with various classes of antidepressant drugs. Recent studies have focused on the functional interaction between the serotonergic and dopaminergic systems to explain the mechanism of the antidepressant action of SSRIs and 5-HT2 antagonists. PMID- 9864077 TI - Differential response to lithium and carbamazepine in the prophylaxis of bipolar disorder. AB - In a randomized, prospective, multicenter study with an observation period of 2.5 years, the differential prophylactic efficacy of lithium versus carbamazepine was compared in 171 patients fulfilling DSM-IV criteria for bipolar disorder. Serum drug levels were 0.6+/-0.1 mmol/L for lithium and 6.1+/-1.3 microg/mL for carbamazepine. Patients were subdivided into a classical subgroup (bipolar I patients without mood-incongruent delusions and without comorbidity, N = 67) and a nonclassical subgroup including all other patients (N = 104). Classical bipolar patients had a lower rehospitalization rate with lithium than with carbamazepine prophylaxis (p = 0.005). For the nonclassical group, a trend in favor of carbamazepine was found. In the lithium group, there was a positive association between hospitalization rate and number of nonclassical features (bipolar II/not otherwise specified, mood-incongruent delusions, comorbidity; p = 0.035). For carbamazepine, this association was negative (p = 0.033). Analyses including mixed states as an additional nonclassical feature confirmed the results. In conclusion, lithium seems to be superior to carbamazepine in treating classical bipolar cases. Patients with nonclassical features might profit more from prophylaxis with carbamazepine, which seems to have a broader spectrum of activity. PMID- 9864078 TI - Gabapentin does not alter single-dose lithium pharmacokinetics. AB - Lithium (Li) and gabapentin are both exclusively eliminated by renal excretion. When used in combination, a competitive drug-drug interaction could possibly alter Li renal excretion with important clinical implications considering the rather narrow therapeutic index of Li. This study examined the single-dose pharmacokinetic profiles of Li in 13 patients receiving placebo and then steady state gabapentin (mean daily dose: 3,646.15 mg). During both phases, a single 600 mg dose of Li was orally administered with serial Li levels obtained at time zero and at 0.25, 0.5, 1, 2, 3, 4, 8, 12, 24, 48, and 72 hours. The pharmacokinetic parameters assessed were the following: area under the concentration time curve (AUC) for Li, maximal concentration of Li (Li Cmax), and time to reach peak Li concentration (Li Tmax). For patients receiving gabapentin, the mean Li AUC at 72 hours was 9.91+/-3.54 mmol x hr/mL and did not differ significantly from the mean Li AUC of 10.19+/-2.89 mmol x hr/mL for patients receiving placebo. The mean Li Cmax was 0.69+/-0.13 mmol/L for gabapentin patients and did not differ from the mean Li Cmax of 0.72+/-0.15 mmol/L for placebo patients. The mean serum Li Tmax was 1.38+/-0.62 hours for gabapentin patients and did not differ significantly from the mean serum Li Tmax of 1.5+/-0.91 hours for placebo patients. These data indicate that gabapentin treatment at this high therapeutic dose does not cause clinically significant alterations in short-term Li pharmacokinetics in patients with normal renal function. These preliminary data warrant further controlled study in a larger, more heterogenous patient sample and a longer duration of assessment, but they do suggest that these two medications may be administered in combination for the management of bipolar disorder. PMID- 9864079 TI - Buspirone augmentation of antidepressant therapy. AB - Thirty outpatients meeting DSM-III-R or DSM-IV criteria for major depression, single or recurrent episode, and failing to respond to an adequate trial of an antidepressant (>6 weeks at recommended dosage) received buspirone (20-30 mg/day) for 4 or 5 weeks in addition to their existing antidepressant. Of the 22 patients who had buspirone added to their selective serotonin reuptake inhibitor antidepressant regimen (fluoxetine, paroxetine, or citalopram), 59% (13/22) showed complete or partial remission of their depressive symptomatology. Similarly, 63% (5/8) of patients treated with buspirone in addition to clomipramine showed complete or partial remission. The mean score on the Clinical Global Impressions Scale fell by 64% (from 4.7 to 1.7; p < 0.0001) in treatment responders (complete and partial). No serious side effects were observed during combination therapy. Seventy-nine percent (11/14) of initial responders (both complete and partial) who remained on augmentation therapy for at least 4 months were symptom-free at follow-up. Buspirone augmentation may produce marked clinical improvement in depressed patients who are initially unresponsive to standard antidepressant therapy. PMID- 9864080 TI - Multiple-dose pharmacokinetics of clozapine in patients with chronic schizophrenia. AB - The pharmacokinetic parameters of clozapine and its two main metabolites, N desmethylclozapine (norclozapine, active metabolite) and clozapine N-oxide, were evaluated, after oral administration, in 19 patients with chronic schizophrenia. Plasma and red blood cell (RBC) drug concentrations were determined by high performance liquid chromatography. Large interpatient variations in pharmacokinetic parameters of clozapine and its two metabolites were observed. Plasma clozapine concentration peaked, on average, at 2.3 hours. The mean volume of distribution and the total plasma clearance, uncorrected for bioavailability, were 6 L/kg and 38 L/hr, respectively. The terminal elimination half-lives averaged 7.6 hours for clozapine, 13 hours for norclozapine, and 7 hours for the N-oxide metabolite. The mean RBC/plasma concentration ratios were 23, 61, and 81% for clozapine, N-desmethylclozapine, and clozapine N-oxide, respectively. From RBC concentration data, the mean elimination half-lives were 7.6 hours for clozapine, 16 hours for N-desmethylclozapine, and 8 hours for the N-oxide metabolite. The average value for blood clearance of clozapine was 54.7 L/hr. Significant correlations were observed between dose and maximum plasma concentrations and between dose and area under the curve concentrations; these results suggested linear steady-state pharmacokinetics over the range of concentrations studied. PMID- 9864081 TI - Prolongation of QT interval in isolated feline hearts by antipsychotic drugs. AB - Some antipsychotic drugs have been found to prolong the QT interval on electrocardiographic (ECG) recordings, a phenomenon which, when severe, may facilitate the occurrence of complex ventricular arrhythmias such as torsade de pointes. However, the effects of these drugs on the cardiac repolarization process have not been evaluated extensively. This study was designed to examine the potency of five antipsychotic drugs in lengthening the QT interval of the perfused feline heart: haloperidol, risperidone, sertindole, clozapine, and olanzapine. The hearts were infused with increasing concentrations of drugs (0.1 20 micromol/L) for 40-minute intervals at each concentration. ECG recordings were made, with signals amplified and data recorded on a strip chart recorder. Four representative beats from each set of three signal recordings were chosen for QT interval measurement. Our data indicated that all tested drugs prolonged the QT interval in a concentration-dependent manner (p < 0.01). Haloperidol and risperidone were significantly more potent than sertindole, clozapine, and olanzapine (p < 0.001). At a concentration of 0.5 micromol/L over a 40-minute infusion interval, haloperidol lengthened the interval by 26.2+/-0.7%, risperidone by 19.4+/-2.2%, and sertindole by 8.9+/-3.5% (p < 0.05 compared with baseline); clozapine and olanzapine were less potent. Although species differences may limit extrapolation of our findings to humans, the cardiac potassium channels of felines clearly resemble those of humans. This model may serve as the basis for further studies of drug-induced prolongation of the QT interval and precipitation of ventricular arrhythmias. PMID- 9864082 TI - Visual hallucinations associated with fluoxetine and sertraline. PMID- 9864083 TI - Extrapyramidal symptoms after addition of fluvoxamine to clozapine. PMID- 9864084 TI - Case report: possible neuroleptic malignant syndrome associated with olanzapine. PMID- 9864085 TI - Return of menstruation after switching from risperidone to olanzapine. PMID- 9864086 TI - Is there a role for the alpha2 antagonism in the exacerbation of hallucinogen persisting perception disorder with risperidone? PMID- 9864087 TI - Nasal toxicity of CI-959, a novel anti-inflammatory drug, in Wistar rats and Beagle dogs. AB - CI-959, 5-methoxy-3-(1-methylethoxy)-N-1H-tetrazol-5-yl-benzo[b]thio phene-2 carboxamide, an anti-inflammatory agent, was considered for development as a treatment for rhinitis. Two-week topical nasal studies in Wistar rats and Beagle dogs were performed to assess nasal toxicity of CI-959. Rats were given daily doses in the right nostril of 0.05 ml of solutions of varying concentrations (0.5, 2, 10, 20, 30, 60, and 90 mg/ml; doses of 0.08, 0.3, 1.6, 3.2, 4.8, 9.6, and 14.6 mg/kg) of CI-959. Beagle dogs were given daily doses in the right nostril of 0.5 ml of 10, 20, 30, 60, and 90 mg/ml solutions (doses of 0.5, 0.8, 1.2, 2.8, and 3.7 mg/kg) of CI-959. Rats given > or = 60 mg/ml either lost weight or had decreased weight gain. Salivation at dosing was seen in both species. Four sections of nasal cavity were examined from each animal. In rats, 0.5 mg/ml was the "no effect" dose; minimal changes were seen at 2 mg/ ml, and significant changes were dose related in severity at > or = 10 mg/ml in all 4 nasal levels. Degeneration and necrosis of respiratory and olfactory epithelia were minimal to moderate in severity. Adhesions and fibro-osseous proliferation of ethmoturbinates, epithelial hyperplasia, squamous metaplasia, and exudate were also seen. In dogs, 10 mg/ml was the no effect dose; respiratory epithelium was affected at > or =20 mg/ml. Respiratory epithelial degeneration was minimal to mild, with loss of ciliated and goblet cells and thinning of mucosa. Distribution of degeneration increased with increased concentrations. In both species, in accordance with the suggested action of CI-959, infiltration with neutrophils was not significant. CI-959 was locally toxic to nasal cavity respiratory and olfactory epithelia in rats and respiratory epithelium in dogs. PMID- 9864088 TI - Time- and dose-dependent development of potassium bromate-induced tumors in male Fischer 344 rats. AB - Potassium bromate (KBrO3) is a rodent carcinogen and a nephro- and neurotoxicant in humans. KBrO3 is used in cosmetics and food products and is a by-product of water disinfection by ozonization. KBrO3 is carcinogenic in the rat kidney, thyroid, and mesothelium and is a renal carcinogen in the male mouse. The present study was designed to investigate the relationship of time and dose to bromate induced tumors in male Fischer 344 (F344) rats and to provide some insight into the development of these tumors. KBrO3 was dissolved in drinking water at nominal concentrations of 0, 0.02, 0.1, 0.2, and 0.4 g/L and administered to male F344 rats as the sole water source for 12, 26, 52, 78, or 100 wk. Renal cell tumors were present after 52 wk of treatment only in the high-dose group. Mesotheliomas developed after 52 wk of treatment on the tunica vaginalis. Mesotheliomas were present at sites other than the testicle after 78 wk of treatment, indicating that their origin was the testicular tunic. Thyroid follicular tumors were present as early as 26 wk in 1 rat each from the 0.1- and 0.2-g/L groups. The present study can be used as a basis for the determination of dose-time relationships of tumor development for a better understanding of KBrO3-induced cancer. PMID- 9864089 TI - Systemic and histopathologic changes in Beagle dogs after chronic daily oral administration of synthetic (ethinyl estradiol) or natural (estradiol) estrogens, with special reference to the kidney and thyroid. AB - ABSTRACT Four groups of 3 male and 3 female sexually mature Beagle dogs were treated daily po with either ethinyl estradiol (EE) or estradiol (E2). A fifth group of 4 males and 4 females acted as a control group. Three groups of dogs were treated with EE: One group was treated at dose levels of 2.0, 1.5, and 1.0 mg/kg for 6 mo; the other 2 groups received either 0.5 mg/kg or 1.0 mg/kg for 1 yr. The fourth group was treated with 5.0 mg/kg E2 for 1 yr. Results obtained for the clinical, hematological, and biochemical parameters and the histopathologic findings of most organs and tissues in EE- and E2-treated dogs were essentially comparable to those documented in the literature for dogs treated with synthetic or natural estrogens. Chronic treatment with EE or E2 induced similar effects, with the exception of mesothelial proliferation of the genital serosa, which was observed in EE-treated dogs only. Additional new estrogen-related findings were observed in the kidneys and thyroid glands of EE- and E2-treated dogs. Increased interstitial fibrous tissue occurred at the corticomedullary junction and in the outer cortex of the kidney. It appeared to originate primarily from the perivascular fibrous tissue of branches of the renal arteries and veins. Extension of this lesion into the renal parenchyma resulted in secondary atrophic changes of tubules and glomeruli. The treatment relationship and specific characteristics of this renal alteration differentiated it from other chronic renal interstitial and vascular diseases. Squamous metaplasia of urogenital tract epithelia, including renal cortical tubule epithelium, occurred as expected in both EE- and E2-treated dogs. Unexpectedly, squamous metaplasia of thyroid follicular epithelium also occurred. It was present in scattered follicles of both EE- and E2-treated dogs. The renal and thyroid changes did not alter clinicopathological function tests for either of these organs. These 2 new findings extend the list of estrogen-related effects in the dog. PMID- 9864090 TI - Use of DNA repair-deficient XPA transgenic mice in short-term carcinogenicity testing. AB - At present (putative) human carcinogens are identified via epidemiological studies and testing using the chronic 2-yr rodent bioassay. Both methods have severe limitations in that they are slow, insensitive, expensive, and are also hampered by many uncertainties. The development of methods to modify specific genes in the mammalian genome has provided promising new tools for use in identifying carcinogens and characterizing their (qualitative) risk. Several transgenic mouse lines are currently under study to test their possible use in short-term carcinogenicity testing. One such candidate alternative transgenic model is the XPA knock-out mouse. These mice have an almost complete deficiency in DNA nucleotide excision repair (NER). Nevertheless, XPA-deficient mice are viable and have a background of a low incidence of spontaneous development of cancers. Approximately 15% of the mice develop hepatocellular adenomas (only after 1.5 yr). After treatment with ultraviolet-B radiation or 7,12 dimethylbenz(a)anthracene, the XPA-deficient mice developed squamous cell carcinomas and papillomas, respectively, on their skin. Oral treatment of XPA deficient mice with benzo[a]pyrene (B[a]P), 2-acetylaminofluorene (2-AAF), and 2 amino-1-methyl-6-phenylimidazo [4,5-b]-pyridine (PhIP) resulted in lymphomas (B[a]P), liver and bladder tumors (2-AAF), and intestinal adenomas plus lymphomas (PhIP). These results look encouraging, but it should be noted that the compounds and agents tested thus far have all been substrate for nucleotide excision repair. Animal studies with different genotoxic or nongenotoxic compounds, as organized for instance within the framework of the International Life Sciences Institute/Health and Environmental Sciences Institute program, are needed to further evaluate the suitability of the XPA model for short-term carcinogenicity testing. PMID- 9864091 TI - Evaluation of the Emu-pim-1 transgenic mouse model for short-term carcinogenicity testing. AB - The value of the chronic rodent carcinogenicity assay in adequately predicting cancer risk in humans has become a matter of debate over the past few years. Therefore, more rapid and accurate alternative tests are urgently needed. Transgenic mouse models, those harboring genetic changes that are relevant to the multistage cancer process, may provide such alternative tests. Transgenic Emu-pim 1 mice, developed by Berns and coworkers in 1989, contain the pimn-1 oncogene, which is expressed at elevated levels in their lymphoid compartments. As a result, these mice are predisposed to the development of T-cell lymphomas. Because of the low incidence of spontaneous tumors and the increased sensitivity to N-ethyl-N-nitrosourea-induced carcinogenesis, Emu-pim-1 mice were suggested to be one of the first potential and attractive candidates to be used in short-term carcinogenicity testing. In the present article, the results from 2 recent short term assays (with mitomycin C and x-rays) are briefly presented, together with a review of all 11 performed bioassays and their corresponding histopathologic and molecular data. The overall results allow the first evaluation of the Emu-pim-1 mouse model with regard to its usefulness in short-term carcinogenicity testing. It has been shown that the model is primarily suitable as a sensitive short-term assay for genotoxic carcinogens that not only induce (at least) gene mutations and/or large deletions and rearrangements but that also sufficiently target the lymphoid system. However, the Emu-pim-1 mice lack sufficient sensitivity to justify their routine use in short-term carcinogenicity testing in general. PMID- 9864092 TI - Transgenic mouse models for the identification of human carcinogens: a European perspective. PMID- 9864093 TI - The proliferation in uterine compartments of intact rats of two different strains exposed to high doses of tamoxifen or toremifene. AB - Uterine Cell proliferation was studied in intact Sprague-Dawley (SD) and Fischer 344 (F344) rats exposed to the antiestrogens tamoxifen (TAM; 5, 10, 20, or 40 mg/kg) and toremifene (TOR: 21.2 or 42.4 mg/kg). The antiestrogens were administered to animals via gavage daily for 2 or 12 wk. Uterine proliferation was assessed using markers for the proliferating cell nuclear antigen (PCNA) and by the bromodeoxyuridine (BrdU) method. Diethylstilbestrol (DES) was used as an estrogenic reference compound. The antiestrogens either reduced or prevented changes of myometrial and stromal proliferation indices (PI). TAM and TOR caused a time-dependent reduction of endometrial glands without an associated decrease in cell proliferation. In the luminal columnar epithelium, the antiestrogens depressed PCNA PI but enhanced BrdU PI, indicating a low continuous DNA synthesis in otherwise quiescent cells. The antiestrogens induced focal hyperplastic multilayered epithelia with PCNA-positive basal cells along segments of the luminal uterine epithelium. We suggest that this hyperplastic epithelium represents remnants from the glandular epithelium. DES was less efficient in inducing these changes but induced squamous metaplasias in the F344 rats. Uterine effects of the 2 antiestrogens were comparable with the exception of I TAM exposed (40 mg/kg) SD rat that showed squamous metaplasia. F344 rats were more sensitive to the estrogenic action of DES than were the SD rats. PMID- 9864094 TI - Histogenetic stereological reconstruction of rat basophilic, clear, and oncocytic neoplastic renal cell lesions using carbonic anhydrase type II-PAS double-stained sections. AB - The histogenesis of 3 types of rat renal cell tumors (basophilic cell, clear cell, and oncocytic) was stereologically analyzed, with particular attention paid to transitions from normal tubules. Early nitrosamine-induced preneoplastic lesions, including dysplastic tubules (altered tubules), epithelial hyperplasias, and small adenomas, were reconstructed using serially sectioned specimens processed for carbonic anhydrase type II (CA) and periodic acid-Schiff (PAS) (CA PAS) double staining to allow easier distinction of the nephron segments: Proximal tubules had a PAS-positive brush border and were weakly positive for CA in the cytoplasm; distal tubules were PAS negative and weakly positive for CA; collecting ducts were PAS negative and strongly positive for CA. Similarly, cytochrome c oxidase (CytOx) and CytOx-PAS double staining was also applied to confirm the character of oncocytic lesions. All basophilic lesions (7 of 7) showed transition to proximal tubules. Clear cell lesions positive for CA, on the other hand, showed transition to distal tubules in 4 of 9 (44.4%) lesions and to collecting ducts in 4 of 9 (44.4%) lesions, but in only 1 of 9 (11%) to a proximal tubule. All oncocytic lesions (16 of 16), characterized by positivity for both CA and CytOx, showed transition to collecting ducts. The results indicate that the origins of renal cell neoplasia are proximal tubules for the basophilic cell lesions, either proximal or distal tubules for their clear cell counterparts, and collecting ducts for oncocytic lesions. PMID- 9864095 TI - Development of fumonisin-induced hepatotoxicity and pulmonary edema in orally dosed swine: morphological and biochemical alterations. AB - The fumonisin (FB) mycotoxins induce liver injury in all species but induce fatal pulmonary edema (PE) only in pigs. They inhibit ceramide synthase in the sphingolipid biosynthetic pathway. To study the pathogenesis of PE, we examined the early events in the development of FB-induced PE and hepatotoxicity in pigs. Pigs were fed FB-contaminated culture material at 20 mg fumonsin B1 (FB1)/kg body weight/day. Groups of 4 pigs were to be euthanatized on 0, 1, 2, 3, 4, or 5 days after initial exposure to FB or when PE developed. Pigs developed PE beginning on day 3; none survived beyond day 4. Progressive elevations in hepatic parameters, including serum enzymes, bile acids, total bilirubin, and histologic changes, began on day 2. Early histologic changes in the lung (day 2) consisted of perivascular edema followed by interlobular and peribronchial edema. Ultrastructurally, alveolar endothelial cells contained unique accumulations of membranous material in the cytocavitary network beginning on day 2. Marked elevations in sphinganine, sphingosine, and their ratio began on day 1 for all tissues whether affected morphologically (lung, liver) or not (kidney, pancreas). The membranous material in endothelial cells may be accumulations of sphingoid bases with damage to the cytocavitary network. Thus, FB induces early elevations in sphingolipids and hepatic injury, followed by alveolar endothelial damage, which may be the critical event in the pathogenesis of PE in pigs. PMID- 9864096 TI - Vacuolation in renal tubular epithelium of Cd-1 mice: an incidental finding. AB - Prominent cytoplasmic vacuoles were observed in renal tubular epithelial cells of the outer medulla in several kidneys from test article-dosed mice (Crl:CD-1 (ICR)BR VAF/PLUS) during routine light microscopic (LM) examination. Because the vacuolar change was detected infrequently and was not found in any control mice from that study, it was not clear whether the vacuolation represented a drug induced change. To address this question, kidney sections from mice from multiple unrelated studies were examined by LM for similar vacuolar changes. Vacuolation was seen by LM in 2.3% of the control and 2.8% of the test article-dosed mice. Transmission electron microscopy (TEM) was also performed on kidneys with prominent light microscopic vacuoles in 5 control mice and 2 test article-dosed mice to further characterize the vacuoles. Ultrastructurally, the vacuoles contained fibrillar and finely stipled granular material or membranous whorls. Kidneys from control mice lacking light microscopic evidence of vacuolation had smaller vacuoles containing similar material when examined by TEM. Because vacuoles were present in both control mice and test article-dosed mice, it was concluded that the vacuoles were incidental and unrelated to compound administration. These studies also demonstrated that vacuoles can be expected to be observed by LM examination in 2-3% of Crl:CD-1 (ICR)BR VAF/PLUS, mice. PMID- 9864097 TI - Immunohistochemical analysis of macrophages, myofibroblasts, and transforming growth factor-beta localization during rat renal interstitial fibrosis following long-term unilateral ureteral obstruction. AB - Renal interstitial fibrosis was induced in rats by chronic unilateral ureteral obstruction (UUO). To identify the mechanisms behind the fibrosis, macrophage influx, myofibroblast involvement, and the localization of transforming growth factor-beta (TGF-beta, a fibrogenic cytokine) were investigated immunohistochemically in rats euthanatized at 0 (controls), 3, 6, 9, 12, and 15 days after UUO. The number of alpha-smooth muscle actin-positive myofibroblasts began to increase significantly in the medulla from day 3, and the development of medullary fibrosis was confirmed from day 6 by morphometric analysis. From day 9, papillary fibrosis also developed in association with an increased number of myofibroblasts. These myofibroblasts showed a parallel orientation to the mucosal surface of the pelvis. In the medulla and papilla, from day 6 the number of ED1 (primary antibody)-positive macrophages began to increase significantly. There appeared to be a relationship between macrophage influx and myofibroblast involvement. By contrast, in the cortex there was no marked increase in myofibroblasts nor development of fibrotic tissues, regardless of increased number of macrophages from day 6. Immunohistochemically, no staining for TGF-beta was found in infiltrating macrophages or myofibroblasts. However, TGF-beta was localized on some cortical proximal renal tubules both of normal control and obstructed kidneys in the early stages on days 3, 6, and 9, suggesting that the possible origin of TGF-beta may be renal epithelia. However, the staining intensity for TGF-beta on the renal epithelia tended to be weakened in advanced obstructed kidneys on days 12 and 15. The likely contribution of TGF-beta to the advanced stages of UUO-induced renal fibrosis remains to be determined. PMID- 9864098 TI - Giant cell tumor of bone in an aged Fischer 344 rat. AB - A giant cell tumor (GCT) was detected on the distal end of the femur in a 98-wk old male Fischer 344 rat. The yellowish white mass had expanded, compressing adjacent muscle tissues. The tumor had an osteolytic and relatively homogeneous appearance and was composed of multinuclear giant cells scattered in a mass of mononuclear stromal cells. No osteoid tissue formation was observed. The tumor cells were strongly immunoreactive for ED-1 and some were also positive for alpha smooth muscle actin, suggesting that the tumor originated from the monocyte/macrophage lineage showing myofibroblastic differentiation. This is the first report concerning spontaneous GCT of bone in a rat. PMID- 9864099 TI - Peripheral neuroblastoma in a young Beagle dog. AB - A peripheral neuroblastoma was found in the abdominal cavity of a young male beagle dog. The large tumor mass involved the left kidney and both adrenal glands. Histologically, a major portion of the neoplasm consisted of lobulated sheets of small round cells with hyperchromatic nuclei mixed with polygonal cells and neuropil. Small clusters of polygonal cells with abundant eosinophilic cytoplasm and a trabecular growth pattern were observed adjacent to some of the tumor lobules. Small, round neoplastic cells metastasized to lumbar lymph nodes and also to the adrenal glands. The neoplastic cells were positive for neuron specific enolase, synaptophysin, and neurofilament protein. Electron micrographs revealed intracytoplasmic dense core granules, microtubules, intermediate filaments, and desmosomes in the cytoplasm of the neoplastic cells. PMID- 9864100 TI - Apoptosis in the kidney. AB - Apoptosis is a highly regulated mechanism of cell death. Although apoptosis has a functional role in normal development and tissue homeostasis, aberrant triggering of the process by toxicants may lead to abnormal function or disease. Low level exposures to toxicants that induce apoptosis in kidney may therefore create a critical disturbance in kidney homeostasis, contributing to renal neoplasia or renal disease. In this report, we review the involvement of apoptosis in normal kidney development and in renal disease and discuss some of the toxicants and molecular factors involved in regulation of the process in renal cells. PMID- 9864101 TI - Apoptosis in the kidney: a toxicologic pathologist's perspective. PMID- 9864102 TI - The 1996 Food Quality Protection Act--an evolution or a revolution for toxicology and risk assessment? PMID- 9864103 TI - Antigen retrieval methods for immunohistochemistry. PMID- 9864104 TI - Laparoscopic hiatal herniorrhaphy with posterior fundoplication for gastroesophageal reflux. AB - Complications and side effects following laparoscopic antireflux procedures are common. This article describes an alternative laparoscopic technique to prevent gastroesophageal reflux. This method consists of posterior approximation of the diaphragmatic crura followed by a posterior fundoplication of approximately 270 degrees wrap. In avoiding the 360 degree wrap and obtaining length from the longitudinal axis of the stomach, it is not necessary to take down the gastrosplenic vessels. The principle of the procedure is to accentuate the cardioesophageal angle of His. No sutures are placed in the esophagus. While this article primarily concerns technique, it also constitutes a brief report on the first 50 patients who have been followed up for 1 year or more. All patients but one are free of reflux symptoms and have discontinued taking all medication. There has been no dysphagia to liquids, and solid food dysphagia has not lasted >1 month. Bloating from gas is minimal, as most patients are able to burp early in their recovery. An outcome paper describing preoperative and postoperative objective testing and evaluation is in process. PMID- 9864105 TI - Gastric endoluminal treatment of Dieulafoy's ulcer. AB - Dieulafoy's ulcer, accounting for approximately 5% of upper GI bleeding, is best managed by therapeutic endoscopy. Some patients, however, require surgical treatment. Presented here is a case in which the lesion was treated by combined endoscopy and endogastric surgery. To the authors' best knowledge, this is the first published report of such management. This relatively new technique may be offered in suitable cases but should be attempted only by well-trained laparoscopic surgeons. PMID- 9864106 TI - Intraperitoneal normal saline and bupivacaine infusion for reduction of postoperative pain after laparoscopic cholecystectomy. AB - After laparoscopic cholecystectomy, CO2 remains within the peritoneal cavity, commonly causing pain. This prospective randomized study was performed to determine the efficacy of intraperitoneal normal saline and bupivacaine infusion on postoperative pain after laparoscopic cholecystectomy. Three hundred patients were randomly assigned to one of six groups of 50 patients each. Group A patients served as controls. In group B patients, normal saline was infused under the right hemidiaphragm and suctioned after the pneumoperitoneum was deflated. After suction, a subhepatic closed drain was left for 24 h. In group C patients, bupivacaine 1.5 mg/kg in solution 2.5 mg/ml, minus 15 ml of this solution, which was infiltrated in the trocar wounds, was infused under the right hemidiaphragm at the end of the cholecystectomy. In group D patients, bupivacaine was given as in group C, but a subhepatic drain was left for 24 h. In group E patients, normal saline was used as in group B plus bupivacaine as in group C. Group F patients were treated as in group E, but a subhepatic drain was left for 24 h. In all groups, 15 ml of a 2.5 mg/ml bupivacaine solution was infiltrated in the trocar wounds. Postoperatively, analgesic medication usage, nausea, vomiting, and pain scores were recorded at 2, 6, 12, 24, 36, 48, and 72 h. Postoperative pain was reduced significantly in the patients of the treatment groups vs. the controls. Between treatment groups, patients in groups B, E, and F had the best results, while those in groups C and D had significantly greater pain than those in groups B, E, and F. It is concluded that postoperative pain after laparoscopic cholecystectomy can be significantly reduced by intraperitoneal normal saline infusion subdiaphragmatically and after its postdeflation suction, bupivacaine infusion in the same area, or without bupivacaine in case a subhepatic drainage has been needed. PMID- 9864107 TI - Laparoscopic cholecystectomy using ultrasonically activated coagulating shears. AB - Ultrasonic energy has recently been used for surgical cutting and coagulating. A prospective randomized study was undertaken to determine the effectiveness of ultrasonic energy versus monopolar electrosurgery in human laparoscopic cholecystectomy. Two hundred patients were enrolled and randomized into two groups of 100 patients each. Group A patients underwent laparoscopic cholecystectomy with monopolar electrocautery. Group B patients underwent laparoscopic cholecystectomy with ultrasonically activated shears. In 18 cases of this group, the cystic artery was coagulated and cut without clips. Subhepatic closed drainage was left for 24 h in patients who were candidates for oozing of blood or leakage of bile. The median operating time was 45 min in group A and 37 min in group B. Subhepatic drainage was left in 37 patients of group A and 26 of group B. The median blood loss was 14 ml in group A and 2 ml in group B, while 3 patients of group A and none of group B had bile leakage from the bed of the gallbladder for 1, 1, and 6 days, respectively. Postoperative ultrasound examination showed a minor subhepatic fluid collection in 5 patients of group A and in 1 patient of group B. All these collections were treated without drainage. The length of hospital stay was 1.9 +/- 0.5 days in group A and 1.4 +/- 0.2 days in group B. Postoperative pain scores, nausea, and vomiting were equivalent in both groups. It is concluded that ultrasonically activated coagulating shears are safer, easier to use, faster, and less prone to intraoperative complications and postoperative morbidity than monopolar electrocautery in laparoscopic cholecystectomy. PMID- 9864108 TI - Laparoscopic approach to the management of incarcerated hernia of appendices epiploicae: report of two cases and review of the literature. AB - Appendices epiploicae may be involved in several intraabdominal pathologic processes, including occasional incarceration in hernias. This diagnosis is most frequently made intraoperatively. In addition to repair of the hernia, management includes a thorough evaluation of the involved appendage in terms of its viability. Any appendage found to be nonviable or of doubtful viability needs to be excised. Such evaluation and excision has traditionally required an open approach. Laparoscopy offers the advantage of a comparable evaluation of viability, the ability to excise the affected appendage if necessary, and a significantly lower morbidity. The requirement of general anesthesia and the need to generate a pneumoperitoneum may be limiting factors in patient selection. PMID- 9864109 TI - Laparoscopic Nissen fundoplication: a prospective analysis of 200 consecutive patients. AB - Nissen fundoplication is now the most common antireflux operation for gastroesophageal reflux disease. This study is a report on the laparoscopically performed floppy Nissen procedure. Two hundred consecutive patients were analyzed (84 women, 116 men, mean age 49 years, mean duration of symptoms 5 years) after laparoscopic Nissen fundoplication between 1992 and 1996. The main indications for surgery were daily heartburn, retrosternal pain, and regurgitation demanding continuous medical therapy. Eight patients (4%) had esophageal stricture, and 21 (11%) had Barrett's esophagus with intestinal metaplasia. All patients underwent upper gastrointestinal endoscopy, 24-h esophageal pH monitoring, and esophageal manometry before and 3 months after the operation. In addition, a questionnaire was completed an average of 2.2 years (range 1.0-4.6) after the operation. The results of the study were as follows: mortality was zero, and the morbidity rate was 5%. The mean hospital stay was 3.8 +/- 2.8 days, and sick leave was 14.3 +/- 10.4 days. Postoperatively, esophagitis was healed or significantly improved in all but 4 patients (98%), and 24-h pH and lower esophageal sphincter pressure were normal. After 2 years, 87% of the patients had Visick scores of I-II. It is concluded that laparoscopic floppy Nissen fundoplication provides an efficient and safe alternative for surgical treatment of gastroesophageal reflux disease. PMID- 9864110 TI - Stress hyperglycemia in minimally invasive surgery. AB - This study examined the selected hormonal responses to, and hormone-mediated glucose metabolism during minimally invasive surgery in, patients undergoing laparoscopic cholecystectomy for symptomatic gallstone disease. Thirty-two patients with symptomatic gallstone disease were included in this study and scheduled for open or laparoscopic procedure in a randomized trial. Results are expressed as mean and standard error of the mean. Statistical evaluations were performed with Mann-Whitney U and Wilcoxon signed-rank tests. Blood cortisol, glucagon, insulin, and glucose concentrations were measured immediately in the preoperative period and 6 h after surgery. Blood cortisol, glucagon, and glucose concentrations increased significantly after open and minimally invasive surgery, while insulin levels and the insulin:glucagon ratio remained unchanged. The rise of glucagon and cortisol values was found to be significantly higher in the postoperative period of the open procedure, than in the laparoscopic approach. However, in the patients who underwent open surgery, the increase in glucose concentrations was not significantly higher in the postoperative period. Surgery induced hormonal effects on the islets increase glucagon and suppress insulin secretion. The glucagon-mediated increase in hepatic glucose production is excluded by the posttraumatic insulin levels from the insulin-sensitive tissues. A bihormonal setting favors a greater rate of hepatic glucose production in both open and laparoscopic surgery. Hormonal changes do reflect the degree of surgical stress, but their metabolic consequences are not parallel to the grade of surgical trauma in minimally invasive surgery. PMID- 9864111 TI - Value and limits of endorectal ultrasonography for preoperative staging of rectal carcinoma. AB - In rectal cancer, the depth of tumor infiltration and metastatic involvement of lymph nodes are important prognostic factors. Endosonography of the rectum, combining the advantages of both endoscopy and sonography, provides information not available from other imaging diagnostic techniques. From January 1989 to December 1997, 85 patients affected by rectal carcinoma were submitted to preoperative evaluation with endorectal ultrasonography. In 75 cases the results obtained with the endosonography were compared to the histology of the resected specimens. Overall accuracy in staging depth of infiltration was 90.7%. Overstaging occurred in 4% of patients, whereas understaging occurred in 5.3%. In staging lymph nodal involvement, overall accuracy was 76%, sensitivity was 69.8%, specificity was 84.4%, positive predictive value was 85.7%, and negative predictive value was 67.5%. Endorectal ultrasound is a safe and accurate diagnostic method for staging both tumor invasion and lymph node metastatic involvement, and for selecting an appropriate surgical strategy in patients affected by rectal cancer. PMID- 9864112 TI - Conservative laparoscopy in the treatment of posttraumatic splenic laceration using microfiber hemostatic collagen: three case histories. AB - Three cases of posttraumatic splenic laceration in young patients are reported. In each case, hemostasis of the lesion was obtained by the use of microfiber collagen in emergency laparoscopy. In two cases, complete hemostasis was achieved and the entire organ was saved. In the third case, splenectomy proved necessary after renewed bleeding occurred. PMID- 9864113 TI - Response of cerebral oxygen metabolism in the head-up position during laparoscopic cholecystectomy. AB - To clarify the influence of the head-up position on cerebral oxygen metabolism during laparoscopy with CO2 insufflation in 12 patients who underwent laparoscopic cholecystectomy, changes in the concentrations of cerebral oxyhemoglobin (HbO2), deoxyhemoglobin (Hb), total hemoglobin (total Hb) and oxidized cytochrome aa3 (Cyt aa3) were measured by use of near-infrared laser spectroscopy. Anesthesia was maintained with nitrous oxide (66%), oxygen, and sevoflurane. Pneumoperitoneum was maintained at an intraabdominal pressure of 10 12 mm Hg by use of CO2. Minute ventilation was adjusted to maintain end-tidal CO2 tension (P(ET)CO2) between 35 and 40 mm Hg during the procedure. Patients were moved from supine to the head-up (20 degree) position before intraabdominal manipulation. The concentration of HbO2 decreased significantly when patients were moved to the head-up position and 30 min thereafter. It remained significantly low after they were returned to the supine position and at the end of surgery. The concentration of Hb was unchanged during the study. Therefore, the concentration of total Hb decreased significantly when patients were moved to the head-up position, as well as 30 min thereafter. It remained significantly low after they were returned to the supine position and at the end of surgery. The concentration of Cyt aa3, however, did not change significantly during the study. These results suggest that the head-up position during laparoscopic cholecystectomy decreases cerebral HbO2 and total Hb. PMID- 9864114 TI - Laparoscopic fenestration in combination with ethanol sclerotherapy prevents a recurrence of symptomatic giant liver cyst. AB - Laparoscopic fenestration in combination with ethanol sclerotherapy of the cyst wall was performed in four patients with symptomatic giant solitary liver cyst. Two of these patients had recurrent liver cysts, which were resistant to simple laparoscopic fenestration. After the combination therapy, none of the cysts recurred during follow-up periods ranging from 11 to 33 months. PMID- 9864115 TI - Retrieval of specimens in laparoscopy using reclosable zipper-type plastic bags: a simple, cheap, and useful method. AB - Surgical specimens must often be extracted during laparoscopic surgery. Although the technologic advances in this field are amazing, simple measures may sometimes work even better than very sophisticated instruments. This is true of the reclosable plastic bags with zipper-type closure that we use for retrieving surgical specimens from the abdominal cavity in order to protect it as well as the abdominal wall. The bags are cheap, offer no problems for sterilization, are easy to obtain, and are available in many sizes. They are very simple to handle, making it easy to slip in the specimen and then extract it from the abdominal cavity. We describe our experience with these bags and a technique for manipulating them. PMID- 9864116 TI - Comparison of laparoscopic rectopexy with open technique in the treatment of complete rectal prolapse: clinical and functional results. AB - The aim of this retrospective study was to compare the functional and clinical results of laparoscopic rectopexy with those of the open technique in two similar groups of patients with complete rectal prolapse and fecal incontinence. Between November 1992 and June 1997, 21 patients underwent abdominal rectopexy. Thirteen patients (group A: 12 women and 1 man, mean age 52.9 years, range 28-70) and 8 patients (group B: 8 women, mean age 58.2 years, range 20-76) were submitted to Well's rectopexy by the open technique and the laparoscopic approach, respectively, without division of the lateral rectal ligaments. Assignment to each group was done randomly. Before the operation, a detailed clinical history was taken, and patients were studied with inspection and digital examination of the anorectum, proctosigmoidoscopy, determination of pancolonic transit time, dynamic defecography, anorectal manometry, and anal electromyography. After the operation, all patients underwent perineal physiotherapy, external electric stimulation, and perineal biofeedback. The mean follow-up time was 29.5 months (range 6-54) in group A and 25.7 months (range 8-45) in group B. Values were compared by chi-square, Mann-Whitney U, and Wilcoxon tests, as appropriate; differences were considered significant at p < 0.05. In both groups, dyschezia and fecal incontinence improved significantly (p < 0.05) after the operation. Basal pressure of anal sphincter, squeezing pressure, and rectoanal reflex improved without significance, whereas anoperineal pain was not significantly reduced. In group B, the postoperative hospital stay was shorter than in group A, with a marked reduction of costs. Laparoscopic Well's rectopexy has the same clinical and functional results as the open technique, with a shorter postoperative hospital stay and lower costs. PMID- 9864117 TI - Technical feasibility of video-assisted lobectomy with radical lymphadenectomy for primary lung cancer. AB - The morbidity and mortality for video-assisted curative resection of lung cancer was evaluated retrospectively. Forty-one consecutive patients with stage I and II lung cancer underwent video-assisted curative lobectomy with complete hilar and mediastinal lymphadenectomy. Conversion to an open procedure was necessary in two patients. The operating times for the second half of the series were shorter than for the first half. Compared with patients receiving a standard open procedure, the video-assisted patients experienced satisfactory results. We conclude that video-assisted curative lobectomy with complete lymphadenectomy for stage I and II lung cancer is technically feasible in the majority of patients, although follow-up is required to determine the long-term prognosis. Comparative series between video-assisted and open procedures should not be conducted until the surgeon has acquired the necessary video-assisted skills. A prospective randomized trial will determine the actual value of video-assisted procedure for lung cancer treatment. PMID- 9864118 TI - Revision of ventriculo-peritoneal shunt under laparoscopic guidance in patients with hydrocephalus. AB - Ventriculo-peritoneal (V-P) shunt is a common treatment for hydrocephalus. However, shunt insufficiency due to obstruction, dislocation, and detachment of the peritoneal tube is frequently encountered. We designed a new technique in which the peritoneal tube is inserted into a target site of the abdominal cavity under laparoscopic guidance. We operated on 9 patients with hydrocephalus using this technique. Shunt insufficiency of the peritoneal tube had been observed in all patients, and 7 patients had undergone repeated revision of the peritoneal tube before this procedure. Our laparoscopic-assisted method ensured tube insertion into the appropriate site, and minimized the abdominal wound and postoperative adhesions. This method also made it possible to differentiate abdominal pain induced by shunt failure from pain caused by other diseases, including appendicitis. In conclusion, we consider this new technique for V-P shunt performed under laparoscopic guidance to be very useful. PMID- 9864119 TI - Ileal volvulus on internal hernia following left laparoscopic-assisted hemicolectomy. AB - Described here is a case of constriction of the ileal loop caused by a volvulus forming on an internal hernia through the mesenteric opening in the anastomotic colonic stumps, following left laparoscopic-assisted hemicolectomy. This experience indicates the need for closure of mesenteric gaps with a continuous suture after laparoscopic-assisted left hemicolectomy. PMID- 9864120 TI - Open vs. laparoscopic repair of perforated peptic ulcer. PMID- 9864122 TI - The power of words. PMID- 9864121 TI - Three-trocar laparoscopic choleystectomy: a continued debate. PMID- 9864123 TI - Wherever your calling takes you. PMID- 9864124 TI - SAEM Kennedy Lecture: medical care for the poor. Society for Academic Emergency Medicine. PMID- 9864125 TI - Keynote address: New England Research Directors Regional Scientific Meeting. PMID- 9864126 TI - Prevalence of aspirin use among patients calling 9-1-1 for chest pain. AB - OBJECTIVE: Early aspirin administration during an acute myocardial infarction (AMI) decreases morbidity and mortality. This investigation examined the extent to which patients with a complaint of chest pain, the symptom most identified with AMI by the general population, self-administer aspirin before the arrival of emergency medical services (EMS) personnel. METHODS: In this prospective, cross sectional prevalence study, data were derived through the analysis of EMS incident reports for patients with a complaint of chest pain from June 1, 1997, to August 31, 1997. RESULTS: The study included 694 subjects. One hundred two (15%) took aspirin for their chest pain before the arrival of EMS personnel. Of the 322 subjects who reported taking aspirin on a regular basis, 82 (26%) took additional aspirin for their acute chest pain. Only 20 (5%) of the 370 patients who were not using regular aspirin therapy self-administered aspirin acutely (p<0.001). In addition, patients with lower intensity of chest pain (p = 0.03) were more likely to take aspirin for their chest pain. CONCLUSION: Only a relatively small fraction of individuals calling 9-1-1 with acute chest pain take aspirin prior to the arrival of EMS personnel. These individuals are more likely to self-administer aspirin if they are already taking it on a regular basis. It is also possible that they are less likely to take aspirin if their chest pain is more severe. PMID- 9864128 TI - Use of emergency departments by elder patients: a five-year follow-up study. AB - OBJECTIVE: To compare the use of emergency medical care by elders in the United States in 1995 with that previously described for 1990. METHODS: A computerized billing database of 88 EDs in 21 states was retrospectively reviewed for 1995, comparing elder and nonelder patients, estimating national use of emergency medical services by elders, and comparing the 1995 data with previously published results for 1990. RESULTS: From 1990 to 1995, the number of ED visits in the United States increased from 92 million to 100 million. The number of visits made by patients aged 65 years or older increased from 13,639,400 (15%) to 15,666,300 (15.7%), but this increase did not reach statistical significance (p = 0.17). The admission rate for elder ED patients increased from 32% to 46% over the five-year interval (p<0.01). This represents more than 7 million hospital admissions for elder patients in 1995. The rate of intensive care unit (ICU) admission for elders decreased from 7% to 6% over the five-year interval (p = 0.56), compared with 1.3% for nonelder patients for both years. Thirty percent of elder ED patients arrived by ambulance in 1990, compared with 33% in 1995 (p = 0.02). Based on 1995 data, elders comprised 39% of patients arriving by ambulance [odds ratio (OR) 4.75, 95% confidence interval (CI) = 4.71 to 4.79], 43% of all admissions (OR 6.59, 95% CI = 6.54 to 6.64), and 47% of ICU admissions (OR 5.00, 95% CI = 4.91 to 5.09). The comparable ORs in 1990 were 4.4, 5.6, and 5.5, respectively. CONCLUSIONS: From 1990 to 1995, the overall number of ED visits increased. The rate of increase was somewhat greater for elder patients. The use of ambulance services also disproportionately grew among elder patients, as did the rate of hospital admission. The overall rate of ICU admission was stable, but actually fell modestly for elder patients. Of these changes, only the increase in the rate of hospital admission for elders reached statistical significance. PMID- 9864127 TI - The tolerability of lactated Ringer's solution and BSS plus for ocular irrigation with and without the Morgan therapeutic lens. AB - OBJECTIVE: To evaluate two solutions, lactated Ringer's (LR) and a balanced salt solution (BSS Plus, Alcon Laboratories, Ft. Worth, TX), compared with normal saline solution (NSS), for ocular irrigation in healthy adult volunteers with and without the Morgan therapeutic lens (MTL). METHODS: This was a prospective, double-blind, randomized study of healthy volunteers who were at least 18 years of age. Exclusion criteria included the use of analgesics within four hours of the study. A complete ophthalmologic examination was performed at baseline and at the completion of the study. Following randomization and prior to any intervention, baseline discomfort scores were obtained by means of a verbally administered, horizontal, 100-mm, unnumbered analog discomfort scale. Both eyes of each volunteer were irrigated simultaneously for 15 minutes, with additional discomfort scores being recorded every 5 minutes using the same 100-mm, unnumbered analog discomfort scale. A global evaluation to assess the method of irrigation and the solutions used for irrigation was completed by both the physician blinded to the treatment groups and the volunteers. The volunteers were continuously monitored for any adverse effects resulting from the irrigation solutions or MTL. RESULTS: Sixty-three volunteers were recruited into the study, with 61 entered in the final analysis. Age and gender were balanced within each group. There was no significant difference in discomfort scores between the two groups; however, all discomfort scores decreased over time (p = 0.008). A lens solution interaction was identified, with LR being the most tolerated when administered with the MTL. A statistically higher ocular pH difference was seen between the pre- and postirrigation readings for the control eye in volunteers irrigated with MTL (p = 0.046). Analysis of the global evaluations for each group revealed no difference in the distributions of physician and volunteer scores. No adverse event was reported in either group. CONCLUSION: There does not appear to be any difference in discomfort scores between the ocular irrigation fluids when used without the MTL. Overall, the use of the MTL appears well tolerated by healthy, adult volunteers. However, there does appear to be a significant lens solution effect on volunteers' discomfort scores, with LR having significantly lower discomfort scores when used for ocular irrigation with the MTL. The authors conclude that the use of the MTL for ocular irrigation is well tolerated and recommend using LR as the irrigation solution for maximal patient comfort. PMID- 9864129 TI - Abdominal pain in geriatric emergency patients: variables associated with adverse outcomes. AB - OBJECTIVE: To determine the diagnoses and outcomes of geriatric patients with abdominal pain, and to identify variables associated with adverse outcomes. METHODS: Geriatric emergency patients (aged 65 years and older) with a complaint of abdominal pain were participants in this longitudinal case series. Eligible patients were followed by telephone contact and chart review, to determine outcomes and final diagnoses. RESULTS: Of 380 eligible patients, follow-up information was available for 375 (97%), for the two months following the ED visit. Final diagnoses included infection (19.2%), mechanical-obstructive disorders (15.7%), ulcers/hypersecretory states (7.7%), urinary tract disease (7.7%), malignancy (7.2%), and others. Although 5.3% of the patients died (related to presenting condition), most (61.3%) patients ultimately recovered. Surgical intervention was required for 22.1% of the patients. Variables associated with adverse outcomes (death, and need for surgical intervention) included hypotension, abnormalities on abdominal radiography, leukocytosis, abnormal bowel sounds, and advanced age. Most physical examination findings were not helpful in identifying patients with adverse outcomes. This study demonstrated a higher incidence of malignancy (7.2%) and a lower incidence of disease necessitating surgical intervention (22.1%) than previously reported. CONCLUSIONS: The majority of geriatric emergency patients with abdominal pain have significant disease necessitating hospital admission. Morbidity and mortality among these patients are high, and specific variables are strongly associated with death and the need for surgical intervention. Absence of these variables does not preclude significant disease. Physical examination findings cannot reliably predict or exclude significant disease. These patients should be strongly considered for hospital admission, particularly when fever, hypotension, leukocytosis, or abnormal bowel sounds are present. PMID- 9864130 TI - Severe sepsis in the emergency department and its association with a complicated clinical course. AB - OBJECTIVE: Infection severity as determined by clinical criteria has been recently classified and studied in hospitalized inpatients. The objective of the study was to use modified criteria to determine the clinical course associated with three levels of infection severity in infected patients admitted from the ED. METHODS: This was a retrospective cohort study involving all patients 18 years of age and older admitted through the ED of an urban teaching hospital during a four-month period whose primary reason for requiring hospitalization was an infection that was recognized in the ED. ED records were reviewed for criteria used to classify patients by three levels of infection severity: no systemic inflammatory response syndrome, sepsis, and severe sepsis (SS). The relationships between these classifications as well as certain clinical characteristics and any complicated clinical course as measured by death and/or admission to an intensive care unit (ICU), and/or prolonged hospitalization, were analyzed. RESULTS: Of 408 patients entered into the study, 138 (33.8%) fulfilled the criteria of SS in the ED. Patients with SS in the ED had a mortality of only 4.3%, though with an increased risk of dying compared with that of the other groups combined [relative risk (RR) = 11.64, 95% confidence interval (CI) = 1.43 to 96.53], an increased risk of ICU stay (RR = 7.65, 95% CI = 4.08 to 14.36), and an increased risk of prolonged hospitalization (RR = 1.99, 95% CI = 1.38 to 2.88). Although age over 60 years and several comorbid conditions also were identified by univariate analysis as risk factors, multivariate analysis revealed that only SS and diabetes mellitus (DM) were independent predictors of a complicated course. In the authors' institution, the positive predictive value (PPV) of SS for complicated clinical course was 0.48 and the negative predictive value (NPV) of no SS for no complicated course was 0.77. The PPV of [SS + DM] was 0.83, and the NPV of [SS, DM, or both] was also 0.83. CONCLUSION: Although the strongest correlate of a complicated clinical course identified in the ED is SS as defined by the study criteria, its specificity and PPV are low. The mortality of ED patients with SS is much lower than the mortality rates reported for inpatients with SS. SS as defined by the study criteria is too sensitive and therefore lacks utility in the ED setting. PMID- 9864131 TI - 1995-1996 SAEM emergency medicine faculty salary/benefits survey. AB - OBJECTIVE: The Society for Academic Emergency Medicine (SAEM) commissioned an emergency medicine (EM) faculty salary and benefit survey for all 1995 Residency Review Committee in Emergency Medicine (RRC-EM)-accredited programs using the SAEM third-generation survey instrument. Responses were collected by SAEM and blinded from the investigators. POPULATION: Seventy-six of 112 (68%) accredited programs responded, yielding data for 1,032 full-time faculty among the four Association of American Medical Colleges (AAMC) regions. METHODS: Blinded program and individual faculty data were entered into a customized version of Filemaker Pro, a relational database program with a built-in statistical package. Salary data were sorted by 115 separate criteria such as program regions, faculty title, American Board of Emergency Medicine (ABEM) certification, academic rank, years postresidency, program size, and whether data were reported to AAMC. Demographic data from 132 categories were analyzed and included number of staff and residents per shift, number of intensive care unit (ICU) beds, obstacles to hiring new staff, and specific type and value of fringe benefits offered. Data were compared with those from the 1990 and 1992 SAEM and the 1995-96 AAMC studies. RESULTS: Mean salaries were reported as follows: all faculty, $158,100; first-year faculty, $131,074; programs reporting data to AAMC, $152,198; programs not reporting data to AAMC, $169,251. Mean salaries as reported by AAMC region: northeast, $155,909; south, $155,403; midwest, $172,260; west, $139,930. Mean salaries as reported by program financial source: community, $175,599; university, $152,878; municipal, $141,566. CONCLUSIONS: Reported salaries for full-time EM residency faculty continue to rise. Salaries in programs reporting data to the AAMC are considerably lower than those not reporting. The gap between ABEM-certified and non-ABEM-certified faculty continues to widen. Residency trained faculty are now shown to earn more than non-residency-trained faculty. Significant regional differences in salaries have been present in all three SAEM surveys. PMID- 9864132 TI - Strengthening emergency medicine in Poland: a training and partnership model. AB - OBJECTIVE: To report a qualitative evaluation of the Partnerships in Health Care/EMS Project between Poland and the United States. The goal of the partnership was to strengthen the emergency medical services (EMS) system in three Polish cities, Krakow, Bialystok, and Lodz. METHODS: The Polish participants were interviewed in Poland approximately eight months after a U.S. based training program. They were asked to evaluate the effectiveness of the partnership project and discuss their experiences incorporating U.S. emergency medicine (EM) knowledge and technology in the Polish EMS system. RESULTS: The Polish physicians identified three major factors that had the greatest impact on the implementation of U.S. EM knowledge in Poland. These factors were the substantive differences between Polish and U.S. EM knowledge and technology, staffing differences in Polish and U.S. ambulances, and the differing role the EMS system plays in the delivery of primary care in the two countries. CONCLUSIONS: The Polish physicians succeeded in training EM providers in the three cities, thus strengthening clinical skills of EMS providers. They also were able to adapt the principles of U.S. EM that they had learned to fit the specific circumstances that characterize Polish emergency care. As in the United States, the health care system in Poland is inseparable from the social, political, and economic realities of the nation. PMID- 9864133 TI - Professionalism in emergency medicine. SAEM Ethics Committee. Society for Academic Emergency Medicine. AB - The Society for Academic Emergency Medicine (SAEM), with the support and participation of the American Board of Emergency Medicine (ABEM), the Council of Residency Directors (CORD), the American College of Emergency Physicians (ACEP), the Emergency Medicine Residents Association (EMRA), the American Academy of Emergency Medicine (AAEM), and the Association of Academic Chairs of Emergency Medicine (AACEM), initiated a project entitled Professionalism in Emergency Medicine. Its concepts were developed by the SAEM Ethics Committee, and are intended to describe proper behaviors and attitudes of the successful practitioner of emergency medicine. The behaviors described are not primarily scientific or technical, since those are defined by the core curriculum for residency training and are tested through certification examinations. This document identifies attitudes and behaviors that enhance trust by placing the patient's interest above other interests. This concept serves as the operative definition of professionalism. The purpose of this article is to clarify the professional attitudes and knowledge that are important to the emergency physician (EP). While no physician is likely to meet idealized standards, all EPs must meet basic standards while striving for the ideal. Awareness of these standards must begin early in the socialization process of emergency medical professionals. The standards must be integrated into residency training as well as the clinical practice of all EPs. PMID- 9864134 TI - Patients with alcohol problems in the emergency department, part 1: improving detection. SAEM Substance Abuse Task Force. Society for Academic Emergency Medicine. AB - Medical and social problems related to alcohol use are frequently seen in the ED. Often, the tempo of emergency medicine practice seems to preclude assessment beyond that required by the acute complaint. However, detection of ED patients with alcohol problems can occur using brief screening tools. This article was developed by members of the SAEM Substance Abuse Task Force, and describes screening tools that have been used successfully to identify at-risk and dependent drinkers. Their brevity, reproducibility, and accuracy vary somewhat, but screening can be realistically performed in the busy ED setting. The early detection of patients with alcohol problems would provide the opportunity for early intervention, and may reduce subsequent morbidity and mortality in this patient population. PMID- 9864135 TI - Patients with alcohol problems in the emergency department, part 2: intervention and referral. SAEM Substance Abuse Task Force. Society for Academic Emergency Medicine. AB - Early intervention and appropriate referral of patients with alcohol problems have the potential to reduce alcohol-related morbidity and mortality. Part 1 of this series introduced screening tools that can be applied in the ED to allow early detection of at-risk drinkers. This article was developed by members of the SAEM Substance Abuse Task Force and describes assessment and intervention techniques once the at-risk or dependent drinkers has been identified. Appropriate aftercare and referral of patients found to have alcohol problems are also discussed. PMID- 9864136 TI - Research fundamentals: III. elements of a research protocol for clinical trials. AB - A clinical trial is a powerful technique for evaluating the effectiveness of an experimental intervention. The initial stages of planning a clinical trial involve choosing and refining a research question, selecting a study design, and deciding on appropriate statistical tests and sample sizes. The success of the study depends upon how well these issues are thought out in advance, and how they can be put into practice. The protocol is the written document that allows the investigator to communicate details of how the research question will be answered. In the following article, the basic components of the research protocol are described. Issues related to quality control, data entry, and pilot testing are discussed. This is the third in a series of research fundamental concept papers, written by members of the SAEM Research Committee. PMID- 9864137 TI - Pericranial injection of local anesthetics for the management of resistant headaches. PMID- 9864138 TI - Meta-analyses of treatment for occult pneumococcal bacteremia. PMID- 9864139 TI - Hyperbaric oxygenation of mechanically ventilated patients poisoned with carbon monoxide. PMID- 9864140 TI - Src-related protein tyrosine kinases in hematopoiesis. PMID- 9864141 TI - Identification of a novel Stat3 recruitment and activation motif within the granulocyte colony-stimulating factor receptor. AB - Stat3 is essential for early embryonic development and for myeloid differentiation induced by the cytokines granulocyte colony-stimulating factor (G CSF) and interleukin-6 (IL-6). Two isoforms of Stat3 have been identified, (p92) and beta (p83), which have distinct transcriptional and biological functions. Activation of both Stat3 and Stat3beta requires the distal cytoplasmic domain of the G-CSFR, which contains four Tyr at positions 704, 729, 744, and 764. The studies reported here were undertaken to determine which, if any, of these tyrosine residues participated in Stat3/beta recruitment and activation. We showed that Stat3 and Stat3beta were affinity purified using phosphopeptides containing Y704 and Y744 but not by nonphosphorylated peptide analogues or by phosphopeptides containing Y729 and Y764. Complementary results were obtained in studies examining the ability of these peptides to destabilize and inhibit DNA binding of activated Stat3. Both Y704 and Y744 contributed to optimal activation of Stat3/beta in M1 murine myeloid leukemia cells containing wild-type and Y-to-F mutant G-CSFR constructs. Carboxy-terminal to Y704 at the +3 position is Gln; YXXQ represents a consensus Stat3 recruitment and activation motif. Y744 is followed at the +3 position by Cys (C); YXXC, represents a novel motif implicated in the recruitment and activation of Stat3. Modeling of the SH2 domain of Stat3 based on homologous SH2 domains of known structure revealed polar residues whose side chains contact the +3 position. This substitution may confer specificity for the Y704- and Y744-based ligands by allowing H-bond formation between the binding surface and the Gln or Cys found at the respective +3 position. PMID- 9864143 TI - CC chemokine receptors, CCR-1 and CCR-3, are potentially involved in antigen presenting cell function of human peripheral blood monocyte-derived dendritic cells. AB - We examined the potential involvement of two CC chemokine receptors (CCRs), CCR-1 and CCR-3, in the functional activation of granulocyte-macrophage colony stimulating factor (GM-CSF) plus interleukin-4 (IL-4)-generated human peripheral blood monocyte-derived immature dendritic cells (DCs). Flow cytometric analysis showed that CCR-1, CCR-3, CCR-5, and CXC chemokine receptor (CXCR)-4 were expressed on the cell surface of monocyte-derived DCs. Treatment with a monoclonal antibody (MoAb) to either CCR-1 or CCR-3 but not MoAbs to CCR-5 and CXCR-4 abolished chemotactic migration of monocyte-derived DCs. The DCs treated with either the anti-CCR-1 MoAb or anti-CCR-3 MoAb were less efficient than untreated DCs in proliferation of allogeneic T cells (TCs) and TC-derived secretion of interferon-gamma (IFN-gamma). The homotypic aggregation of DCs and heterotypic aggregation of DCs with TCs were suppressed by the anti-CCR-1 MoAb or anti-CCR-3 MoAb. These results indicate that CCR-1 and CCR-3 specifically regulate interaction of TCs and DCs in the process of antigen presentation. PMID- 9864142 TI - Inhibition of human umbilical vein endothelial cell proliferation by the CXC chemokine, platelet factor 4 (PF4), is associated with impaired downregulation of p21(Cip1/WAF1). AB - Human PF4 is a heparin-binding chemokine known to be capable of inhibiting endothelial cell proliferation and angiogenesis. To explore the biological mechanisms responsible for this action, we investigated the effect of PF4 on epidermal growth factor (EGF)-stimulated human umbilical vein endothelial cells (HUVEC), a model system in which stimulation is essentially independent of interaction with cell-surface glycosaminoglycans. Based on previous findings that PF4 blocks endothelial cell cycle entry and progression into S phase, we studied the molecular mechanism(s) of PF4 interference with cell cycle machinery. PF4 treatment of EGF-stimulated HUVEC caused a decrease in cyclin E-cyclin-dependent kinase 2 (cdk2) activity with resulting attenuation of retinoblastoma protein phosphorylation. PF4-dependent downregulation of cyclin E-cdk2 activity was associated with increased binding of the cyclin-dependent kinase inhibitor, p21(Cip1/WAF1), to the cyclin E-cdk2 complex. Analysis of total cellular p21(Cip1/WAF1) showed that in the presence of PF4, p21(Cip1/WAF1) levels were sustained at time points when p21(Cip1/WAF1) was no longer detectable in cells stimulated by EGF in the absence of PF4. These findings indicate that PF4 inhibition of HUVEC proliferation in response to EGF is associated with impaired downregulation of p21(Cip1/WAF1) and provide the first evidence for interference with cell cycle mechanisms by a chemokine. PMID- 9864144 TI - Murine T lymphocytes incapable of producing macrophage inhibitory protein-1 are impaired in causing graft-versus-host disease across a class I but not class II major histocompatibility complex barrier. AB - The routine use of bone marrow transplantation is limited by the occurrence of acute and chronic graft-versus-host disease (GVHD). Current approaches to decreasing the occurrence of GVHD after allogeneic transplantation use T-cell depletion, use immunosuppressive agents, or block costimulatory molecule function. The role of proteins in the recruitment of alloreactive lymphocytes has not been well characterized. Chemokines are a large family of proteins that mediate recruitment of mononuclear cells in vitro and in vivo. To investigate the role of T-cell production of the chemokine macrophage inhibitory protein-1 (MIP 1) in the occurrence of GVHD, splenocytes either from wild-type or from MIP-1-/- mice were administered to class I (B6.C-H2(bm1)) and class II disparate mice (B6 C-H2(bm12)). The incidence and severity of GVHD was markedly reduced in bm1 mice receiving splenocytes from MIP-1-/- mice as compared with mice receiving wild type splenocytes. Bm1 mice receiving MIP-1-/- splenocytes had significantly less weight loss and markedly reduced inflammatory responses in the lung and liver than mice receiving C57BL/6 splenocytes. Bm1 mice receiving MIP-1-/- splenocytes had a markedly decreased production of antichromatin autoantibodies and impaired generation of bm1-specific T lymphocytes versus wild-type mice. However, MIP-1-/- splenocytes easily induced GVHD when administered to bm12 mice. This data show that blockade of chemokine production or function may provide a new approach to the prevention or treatment of GVHD but that chemokines that recruit both CD4(+) and CD8(+) lymphocytes may need to be targeted. PMID- 9864145 TI - Age is not a prognostic variable with autotransplants for multiple myeloma. AB - Multiple myeloma (MM) typically afflicts elderly patients with a median age of 65 years. However, while recently shown to provide superior outcome to standard treatment, high-dose therapy (HDT) has usually been limited to patients up to 65 years. Among 550 patients with MM and a minimum follow-up of 18 months, 49 aged >/=65 years were identified (median age, 67; range, 65 to 76 years). Their outcome was compared with 49 younger pair mates (median, 52; range, 37 to 64 years) selected among the remaining 501 younger patients (<65 years) matched for five previously recognized critical prognostic factors (cytogenetics, beta2 microglobulin, C-reactive protein, albumin, creatinine). Nearly one half had been treated for more than 1 year with standard therapy and about one third had refractory MM. All patients received high-dose melphalan-based therapy; 76% of the younger and 65% of the older group completed a second transplant (P =.3). Sufficient peripheral blood stem cells to support two HDT cycles (CD34 > 5 x 10(6)/kg) were available in 83% of younger and 73% of older patients (P =.2). After HDT, hematopoietic recovery to critical levels of granulocytes (>500/microL) and of platelets (>50,000/microL) proceeded at comparable rates among younger and older subjects with both first and second HDT. The frequency of extramedullary toxicities was comparable. Treatment-related mortality with the first HDT cycle was 2% in younger and 8% among older subjects, whereas no mortality was encountered with the second transplant procedure. Comparing younger/older subjects, median durations of event-free and overall survival were 2.8/1.5 years (P =.2) and 4.8/3.3 years (P =.4). Multivariate analysis showed pretransplant cytogenetics and beta2-microglobulin levels as critical prognostic features for both event-free and overall survival, whereas age was insignificant for both endpoints (P =.2/.8). Thus, age is not a biologically adverse parameter for patients with MM receiving high-dose melphalan-based therapy with peripheral blood stem cell support and, hence, should not constitute an exclusion criterion for participation in what appears to be superior therapy for symptomatic MM. PMID- 9864146 TI - Total therapy with tandem transplants for newly diagnosed multiple myeloma. AB - Between August 1990 and August 1995, 231 patients (median age 51, 53% Durie Salmon stage III, median serum beta-2-microglobulin 3.1 g/L, median C-reactive protein 4 g/L) with symptomatic multiple myeloma were enrolled in a program that used a series of induction regimens and two cycles of high-dose therapy ("Total Therapy"). Remission induction utilized non-cross-resistant regimens (vincristine doxorubicin-dexamethasone [VAD], high-dose cyclophosphamide and granulocyte macrophage colony-stimulating factor with peripheral blood stem cell collection, and etoposide-dexamethasone-cytarabine-cisplatin). The first high-dose treatment comprised melphalan 200 mg/m2 and was repeated if complete (CR) or partial (PR) remission was maintained after the first transplant; in case of less than PR, total body irradiation or cyclophosphamide was added. Interferon--2b maintenance was used after the second autotransplant. Fourteen patients with HLA-compatible donors underwent an allograft as their second high-dose therapy cycle. Eighty eight percent completed induction therapy whereas first and second transplants were performed in 84% and 71% (the majority within 8 and 15 months, respectively). Eight patients (3%) died of toxicity during induction, and 2 (1%) and 6 (4%) during the two transplants. True CR and at least a PR (PR plus CR) were obtained in 5% (34%) after VAD, 15% (65%) at the end of induction, and 26% (75%) after the first and 41% (83%) after the second transplants (intent-to treat). Median overall (OS) and event-free (EFS) survival durations were 68 and 43 months, respectively. Actuarial 5-year OS and EFS rates were 58% and 42%, respectively. The median time to disease progression or relapse was 52 months. Among the 94 patients achieving CR, the median CR duration was 50 months. On multivariate analysis, superior EFS and OS were observed in the absence of unfavorable karyotypes (11q breakpoint abnormalities, -13 or 13-q) and with low beta-2-microglobulin at diagnosis. CR duration was significantly longer with early onset of CR and favorable karyotypes. Time-dependent covariate analysis suggested that timely application of a second transplant extended both EFS and OS significantly, independent of cytogenetics and beta-2-microglobulin. Total Therapy represents a comprehensive treatment approach for newly diagnosed myeloma patients, using multi-regimen induction and tandem transplantation followed by interferon maintenance. As a result, the proportion of patients attaining CR increased progressively with continuing therapy. This observation is particularly important because CR is a sine qua non for long-term disease control and, eventually, cure. PMID- 9864147 TI - Etretinate therapy for refractory sclerodermatous chronic graft-versus-host disease. AB - Chronic graft-versus-host disease (GVHD) is the most common late complication of allogeneic bone marrow transplantation (BMT). The sclerodermatous form of the disease is often refractory to standard treatment modalities. Based on reports of response to etretinate, a synthetic retinoid, among patients with scleroderma, we have added etretinate to the treatment regimen of 32 patients with refractory sclerodermatous chronic GVHD. This case series is comprised mainly of patients who had chronic GVHD of long duration (median of 30 months before the initiation of etretinate). Most had failed to respond to three or more agents before etretinate treatment was started. Clinical response was assessed after 3 months of therapy. Five patients did not complete a 3-month trial. Among the 27 patients evaluable for response, 20 showed improvement including softening of the skin, flattening of cutaneous lesions, increased range of motion, and improved performance status. Four showed no response after 3 months of therapy and 3 had progression of their sclerosis. Overall, etretinate has been fairly well tolerated in our patients, with skin breakdown and/or ulceration leading to its discontinuation in 6 patients. We believe the results in our patients are encouraging and suggest that further evaluation of etretinate in the treatment of sclerodermatous chronic GVHD is warranted. PMID- 9864148 TI - Cerebral vasculopathy in sickle cell anemia: diagnostic contribution of positron emission tomography. AB - Children with sickle cell anemia (SS) have an increased risk for cerebral vasculopathy with stroke (CVA) and cognitive impairment. The present study examines the extent to which adding positron emission tomography (PET) to magnetic resonance imaging (MRI) can improve the detection of cerebral vasculopathy. Whereas MRI has been the prime modality for showing anatomical lesions, PET excels at assessing the functional metabolic state through glucose utilization 2-deoxy-2 [18F] fluoro-D-glucose (FDG) and microvascular blood flow ([15O]H2O). Forty-nine SS children were studied. Among them, 19 had clinically overt CVA, 20 had life-threatening hypoxic episodes or soft neurologic signs, and 10 were normal based on neurological history and examination. For the entire sample of 49 subjects, 30 (61%) had abnormal MRI findings, 36 (73%) had abnormal PET findings, and 44 (90%) showed abnormalities on either the MRI or the PET or both. Of the 19 subjects with overt CVA, 17 had abnormal MRI (89%), 17 had abnormal PET (89%), and 19 (100%) had either abnormal MRI or PET or both. Among the 20 subjects with soft neurologic signs, 10 (50%) had abnormal MRI, 13 (65%) had abnormal PET, and 17 (85%) had abnormal MRI and/or PET. Six (60%) of the 10 neurologically normal subjects had abnormal PET. Among the 30 subjects with no overt CVA, 25 (83%) demonstrated imaging abnormalities based on either MRI or PET or both, thus, silent ischemia. Lower than average full-scale intelligence quotient (FSIQ) was associated with either overt CVA or silent ischemic lesions. Four subjects who received chronic red blood cell transfusion showed improved metabolic and perfusion status on repeat PET scans. In conclusion, (1) the addition of PET to MRI identified a much greater proportion of SS children with neuroimaging abnormalities, particularly in those who had no history of overt neurologic events. (2) PET lesions are more extensive, often bihemispheric, as compared with MRI abnormalities. (3) PET may be useful in management as a tool to evaluate metabolic improvement after therapeutic interventions, and (4) the correlation of PET abnormalities to subsequent stroke or progressive neurologic dysfunction requires further study. PMID- 9864149 TI - Fluorescence-based selection of gene-corrected hematopoietic stem and progenitor cells from acid sphingomyelinase-deficient mice: implications for Niemann-Pick disease gene therapy and the development of improved stem cell gene transfer procedures. AB - The general utility of a novel, fluorescence-based procedure for assessing gene transfer and expression has been demonstrated using hematopoietic stem and progenitor cells. Lineage-depleted hematopoietic cells were isolated from the bone marrow or fetal livers of acid sphingomyelinase-deficient mice, and retrovirally transduced with amphotropic or ecotropic vectors encoding a normal acid sphingomyelinase (ASM) cDNA. Anti-c-Kit antibodies were then used to label stem- and progenitor-enriched cell populations, and the Bodipy fluorescence was analyzed in each group after incubation with a Bodipy-conjugated sphingomyelin. Only cells expressing the functional ASM (ie, transduced) could degrade the sphingomyelin, thereby reducing their Bodipy fluorescence as compared with nontransduced cells. The usefulness of this procedure for the in vitro assessment of gene transfer into hematopoietic stem cells was evaluated, as well as its ability to provide an enrichment of transduced stem cells in vivo. To show the value of this method for in vitro analysis, the effects of retroviral transduction using ecotropic versus amphotropic vectors, various growth factor combinations, and adult bone marrow versus fetal liver stem cells were assessed. The results of these studies confirmed the fact that ecotropic vectors were much more efficient at transducing murine stem cells than amphotropic vectors, and that among the three most commonly used growth factors (stem cell factor [SCF] and interleukins 3 and 6 [IL-3 and IL-6]), SCF had the most significant effect on the transduction of stem cells, whereas IL-6 had the most significant effect on progenitor cells. In addition, it was determined that fetal liver stem cells were only approximately twofold more "transducible" than stem cells from adult bone marrow. Transplantation of Bodipy-selected bone marrow cells into lethally irradiated mice showed that the number of spleen colony-forming units that were positive for the retroviral vector (as determined by polymerase chain reaction) was 76%, as compared with 32% in animals that were transplanted with cells that were nonselected. The methods described within this manuscript are particularly useful for evaluating hematopoietic stem cell gene transfer in vivo because the marker gene used in the procedure (ASM) encodes a naturally occurring mammalian enzyme that has no known adverse effects, and the fluorescent compound used for selection (Bodipy sphingomyelin) is removed from the cells before transplantation. PMID- 9864150 TI - Definitive but not primitive hematopoiesis is impaired in jumonji mutant mice. AB - A novel gene, jumonji was identified by a mouse gene trap strategy. The jumonji gene encodes a protein containing a putative DNA binding domain. The mice homozygous for jumonji gene with a BALB/cA genetic background show hypoplasia of the fetal liver and embryonic lethality, suggesting impaired hematopoiesis. In the peripheral blood of jumonji mutant embryos, the number of fetal liver-derived definitive erythrocytes, but not yolk sac-derived primitive erythrocytes, showed a marked reduction, suggesting that jumonji mutants die of anemia. The defects of definitive erythrocytes in jumonji mutants seemed to be caused by a decrease in the numbers of multiple hematopoietic progenitors including colony-forming unit spleen (CFU-S) in the fetal liver. However, hematopoietic stem cells (HSCs) in the fetal liver of jumonji mutants could reconstitute the hematopoietic system of lethally irradiated recipients. In the fetal liver, the jumonji gene is expressed in fibroblastic cells and endothelial cells, but not in Lin-/c-Kit+/Sca-1(+) cells known to include HSCs. These results suggest that an environmental defect induce the impaired hematopoiesis in the fetal liver of jumonji mutant embryos. PMID- 9864151 TI - Single adult human CD34(+)/Lin-/CD38(-) progenitors give rise to natural killer cells, B-lineage cells, dendritic cells, and myeloid cells. AB - Marrow stromal cultures support adult CD34(+)/Lin-/HLA-DR- or CD34(+)/Lin-/CD38( ) cell differentiation into natural killer (NK) or myeloid cells, but unlike committed lymphoid progenitors (CD34(+)/Lin-/CD45RA+/CD10(+)), no B cells are generated. We tested whether different microenvironments could establish a developmental link between the NK and B-cell lineages. Progenitors were cultured in limiting dilutions with interleukin-7 (IL-7), flt3 ligand (FL), c-kit ligand (KL), IL-3, IL-2, and AFT024, a murine fetal liver line, which supports culture of transplantable murine stem cells. NK cells, CD10(+)/CD19(+) B-lineage cells and dendritic cells (DC) developed from the same starting population and IL-7, FL, and KL were required in this process. Single cell deposition of 3,872 CD34(+)/Lin-/CD38(-) cells onto AFT024 with IL-7, FL, KL, IL-2, and IL-3 showed that a one time addition of IL-3 at culture initiation was essential for multilineage differentiation from single cells. Single and double lineage progeny were frequently detected, but more importantly, 2% of single cells could give rise to at least three lineages (NK cells, B-lineage cells, and DC or myeloid cells) providing direct evidence that NK and B-lineage differentiation derive from a common lymphomyeloid hematopoietic progenitor under the same conditions. This study provides new insights into the role of the microenvironment niche, which governs the earliest events in lymphoid development. PMID- 9864152 TI - Murine hematopoietic progenitor cells with colony-forming or radioprotective capacity lack expression of the beta 2-integrin LFA-1. AB - Recently, we have demonstrated that antibodies that block the function of the beta2-integrin leukocyte function-associated antigen-1 (LFA-1) completely abrogate the rapid mobilization of hematopoietic progenitor cells (HPC) with colony-forming and radioprotective capacity induced by interleukin-8 (IL-8) in mice. These findings suggested a direct inhibitory effect of these antibodies on LFA-1-mediated transmigration of stem cells through the bone marrow endothelium. Therefore, we studied the expression and functional role of LFA-1 on murine HPC in vitro and in vivo. In steady state bone marrow +/- 50% of the mononuclear cells (MNC) were LFA-1(neg). Cultures of sorted cells, supplemented with granulocyte colony-stimulating factor (G-CSF)/granulocyte-macrophage colony stimulating factor (GM-CSF)/IL-1/IL-3/IL-6/stem cell factor (SCF) and erythropoietin (EPO) indicated that the LFA-1(neg) fraction contained the majority of the colony-forming cells (CFCs) (LFA-1(neg) 183 +/- 62/7,500 cells v LFA-1(pos) 29 +/- 17/7,500 cells, P <.001). We found that the radioprotective capacity resided almost exclusively in the LFA-1(neg) cell fraction, the radioprotection rate after transplantation of 10(3), 3 x 10(3), 10(4), and 3 x 10(4) cells being 63%, 90%, 100%, and 100% respectively. Hardly any radioprotection was obtained from LFA-1(pos) cells. Similarly, in cytokine (IL-8 and G-CSF)-mobilized blood, the LFA-1(neg) fraction, which comprised 5% to 10% of the MNC, contained the majority of the colony-forming cells, as well as almost all cells with radioprotective capacity. Subsequently, primitive bone marrow derived HPC, represented by Wheat-germ-agglutinin (WGA)+/Lineage (Lin)-/Rhodamine (Rho)- sorted cells, were examined. More than 95% of the Rho- cells were LFA 1(neg). Cultures of sorted cells showed that the LFA-1(neg) fraction contained all CFU. Transplantation of 150 Rho- LFA-1(neg) or up to 600 Rho-LFA-1(pos) cells protected 100% and 0% of lethally irradiated recipient mice, respectively. These results show that primitive murine HPC in steady-state bone marrow and of cytokine-mobilized blood do not express LFA-1. PMID- 9864153 TI - Tyrosine-dependent and -independent mechanisms of STAT3 activation by the human granulocyte colony-stimulating factor (G-CSF) receptor are differentially utilized depending on G-CSF concentration. AB - The granulocyte colony-stimulating factor receptor (G-CSF-R) activates multiple STAT proteins. Although the membrane-proximal cytoplasmic region of the G-CSF-R is necessary and sufficient for activation of STAT1 and STAT5, activation of STAT3 requires the membrane distal region that contains four tyrosines. Although one of these (Y704) has previously been shown to be involved in STAT3 activation from a truncated G-CSF-R derived from a patient with severe chronic neutropenia (SCN), this tyrosine is not required for STAT3 activation by the full-length G CSF-R. To investigate possible alternative mechanisms of STAT3 activation, we generated a series of Ba/F3 cell transfectants expressing the wild-type G-CSF-R or mutant receptors that either completely lack tyrosines or retain just one of the four cytoplasmic tyrosines of the G-CSF-R. We show that, at saturating G-CSF concentrations, STAT3 activation from the full-length G-CSF-R is efficiently mediated by the C-terminal domain in a manner independent of receptor tyrosines. In contrast, at low G-CSF concentrations, Y704 and Y744 of the G-CSF-R play a major role in STAT3 activation. Both tyrosine-dependent and -independent mechanisms of STAT3 activation are sensitive to the Jak2 inhibitor AG-490, follow similar kinetics, and lead to transactivation of a STAT3 reporter construct, indicating functional equivalence. STAT3 activation is also impaired, particularly at nonsaturating G-CSF concentrations, in bone marrow cells from mice expressing a truncated G-CSF-R (gcsfr-triangle up715). These findings suggest that G-CSF-induced STAT3 activation during basal granulopoiesis (low G CSF) and "emergency" granulopoiesis (high G-CSF) are differentially controlled. In addition, the data establish the importance of the G-CSF-R C-terminus in STAT3 activation in primary cells, which has implications for understanding why truncated G-CSF-R derived from SCN patients are defective in maturation signaling. PMID- 9864154 TI - Autonomous megakaryocyte growth in essential thrombocythemia and idiopathic myelofibrosis is not related to a c-mpl mutation or to an autocrine stimulation by Mpl-L. AB - Essential thrombocythemia (ET) and idiopathic myelofibrosis (PMF) are two myeloproliferative diseases characterized by a marked megakaryocytic (MK) involvement. The pathogenesis of these two diseases is unknown. Recently it has been shown that overexpression of Mpl-ligand (Mpl-L) in mice induces thrombocytosis and myelofibrosis. In this study, we investigated whether Mpl-L was responsible for the pathogenesis of ET and PMF. Using in vitro cultures of blood or marrow CD34(+) cells, we investigated whether MK growth was abnormal in these two diseases. Spontaneous MK growth involving only a fraction (20%) of the MK progenitors, as compared with growth in the presence of pegylated recombinant human megakaryocyte growth and development factor (PEG-rhuMGDF), was found in both diseases (21ET and 14PMF) using serum-free semisolid and liquid cultures, including cultures at one cell per well. We first searched for a c-mpl mutation/deletion by sequencing the entire coding region of the gene by polymerase chain reaction (PCR) in nine ET patients and five PMF patients, but no mutation was found. We subsequently investigated whether an autocrine stimulation by Mpl-L could explain the autonomous MK growth. Addition of different preparations of soluble Mpl receptor (sMpl) containing a Fc domain of IgG1 (sMpl Fc) markedly inhibited MK spontaneous growth in both ET and PMF patients. This effect was specific for sMpl because a control soluble receptor (s4-1BB-Fc) had no inhibitory effect and an sMpl devoid of the Fc fragment had the same inhibitory efficacy as the sMpl-Fc. This inhibition was reversed by addition of PEG-rhuMGDF or a combination of cytokines. The sMpl-Fc markedly altered the entry into cell cycle of the CD34(+) cells and increased the apoptosis that occurs in most patient CD34(+) cells in the absence of exogenous cytokine, suggesting an autocrine stimulation. In contrast, a neutralizing antibody against Mpl-L did not alter the spontaneous MK growth, whereas it totally abolished the effects of 10 ng/mL PEG-rhuMGDF on patient or normal CD34(+) cells. Mpl-L transcripts were detected at a very low level in the patient CD34(+)cells and MK and only when a highly sensitive fluorescent PCR technique was used. By quantitative reverse transcription (RT)-PCR, the number of Mpl-L transcripts per actin transcripts was lower than detected in human Mpl-L-dependent cell lines, suggesting that this synthesis of Mpl-L was not biologically significant. In favor of this hypothesis, the Mpl-L protein was not detected in culture supernatants using either an enzyme linked immunosorbent assay (ELISA) or a biological (Ba/F3hu c-mpl) assay, except in one PMF patient. Investigation of Mpl-L signaling showed an absence of constitutive activation of STATs in spontaneously growing patient MKs. Addition of PEG-rhuMGDF to these MKs activated STATs 3 and 5. This result further suggests that spontaneous growth is neither related to a stimulation by Mpl-L nor to a c mpl mutation. In conclusion, our results show that Mpl-L or Mpl are not directly implicated in the abnormal proliferation of MK cells from ET and PMF. The mechanisms by which the sMpl mediates a growth inhibition will require further experiments. PMID- 9864155 TI - Heparan sulfate proteoglycans mediate interleukin-7-dependent B lymphopoiesis. AB - Heparin/heparan sulfate proteoglycans (HSPGs) have the potential to bind and directly regulate the bioactivity of hematopoietic growth factors including interleukin-7 (IL-7), a cytokine critical for murine B-cell development. We examined the consequence of manipulating soluble heparin and cell-surface heparan sulfate to IL-7-dependent responses of B-cell precursors. Soluble heparin was found to inhibit production of lymphoid, but not myeloid, cells in long-term bone marrow cultures. Analysis of pro-B cells lacking plasma membrane HS suggests that this glycosaminoglycan is required for efficient binding and responsiveness to IL 7. By contrast, responses of hematopoietic cells to other cytokines were not influenced by heparin addition or HS removal. Therefore, HSPGs on B-lineage precursors may function as IL-7 receptor components similar to HSPGs known to be important for the bFGF receptor. Other experiments suggest that HSPGs on the surface of stromal cells provide a weakly associating docking site for IL-7, possibly controlling availability of this cytokine to B-cell precursors. Together these data demonstrate a direct role for heparinlike molecules in regulating the IL-7-dependent stages of murine B lymphopoiesis. PMID- 9864156 TI - Selective transendothelial migration of hematopoietic progenitor cells: a role in homing of progenitor cells. AB - To elucidate the mechanisms by which hematopoietic progenitor cells transmigrate via the bone marrow (BM) endothelial cells, we first established endothelial cell lines from BM and lung, and BM fibroblast cell lines; then we established an in vitro model of transendothelial migration of hematopoietic progenitor cells in the presence of chemoattractants secreted by BM fibroblast cells. The BM endothelial cells expressed vascular cell adhesion molecule-1 (VCAM-1), but the lung endothelial cells did not. The BM fibroblast cells secreted chemoattractants including stroma cell-derived factor (SDF)-1, which could attract hematopoietic progenitor cells to BM and activate the adhesion molecules expressed on hematopoietic progenitor cells after rolling along the endothelial cells. Anti SDF-1 antibody inhibited the transendothelial migration of a hematopoietic progenitor cell line, FDCP-2. FDCP-2 that expressed very late activation antigen 4 (VLA-4) and normal progenitor cells transmigrated through BM endothelial cells but not lung endothelial cells, even if in the presence of chemoattractants produced by BM fibroblasts. Both anti-VLA-4 and anti-VCAM-1 antibodies inhibited the transendothelial migration of FDCP-2 cells and normal hematopoietic progenitor cells. These findings suggest that the transendothelial migration of hematopoietic progenitor cells is characteristic of BM endothelial cells, and that VLA-4/VCAM-1 and SDF-1 play important roles in the transendothelial migration and, consequently, homing of hematopoietic progenitor cells to BM. PMID- 9864157 TI - Antithrombin reduces ischemia/reperfusion injury of rat liver by increasing the hepatic level of prostacyclin. AB - We investigated whether antithrombin (AT) can reduce ischemia/reperfusion (I/R) induced injury of rat liver by promoting prostacyclin release from endothelial cells. Although intravenous administration of AT (250 U/kg) markedly reduced hepatic injury, neither dansyl-Glu-Gly-Arg-chloromethyl ketone-treated factor Xa (DEGR-Xa), a selective inhibitor of thrombin generation, nor Trp49-modified AT, which lacks affinity for heparin, had any effect. Hepatic levels of 6-keto-PGF1, a stable prostacyclin (PGI2) metabolite, were increased significantly after I/R of the rat liver. AT significantly increased the hepatic level of 6-keto-PGF1, whereas neither DEGR-Xa nor Trp49-modified AT increased it. Hepatic tissue blood flow was markedly reduced after I/R. Although AT significantly increased the hepatic tissue blood flow after I/R, neither DEGR-Xa nor Trp49-modified AT increased the blood flow. Hepatic levels of cytokine-induced neutrophil chemoattractant (CINC) and myeloperoxidase (MPO) were significantly increased after hepatic I/R. The levels of these two indicators were reduced by AT but were unaffected by either DEGR-Xa or Trp49-modified AT. Pretreatment of animals with indomethacin (IM) completely inhibited the protective effects of AT on the I/R induced hepatic damage and the leukocyte activation as well as the AT-induced increase in hepatic 6-keto-PGF1 levels after I/R. Iloprost, a stable analog of PGI2, exhibited effects similar to those of AT and also significantly inhibited the exacerbation of liver injury, the decrease in hepatic tissue blood flow, and the increases in hepatic CINC and MPO levels seen in rats subjected to I/R but pretreated with IM. These findings suggest that AT may prevent I/R-induced hepatic injury by increasing the hepatic levels of PGI2 through the interaction of AT with cell-surface glycosaminoglycans, thus increasing hepatic tissue blood flow and inhibiting leukocyte activation in animals subjected to I/R. PMID- 9864158 TI - Biochemical characterization and molecular cloning of a novel endothelial specific sialomucin. AB - We have generated rat monoclonal antibodies (MoAbs) against cell surface antigens of the mouse endothelioma cell line bEND.3. Three antibodies (V.1A7, V.5C7, and V.7C7) were selected, all of which recognize a 75-kD antigen on bEND.3 cells and bind selectively to endothelial cells in cryostat sections of mouse tissues. A cDNA for the antigen was isolated from a bEND.3 pCDM8 expression library by using transient expression in COS-7 cells and immunoselection with the three MoAbs. This cDNA coded for a novel, type I membrane protein of 248 amino acids with an extracellular domain rich in threonine and serine residues (35%). The protein is sensitive to O-sialoglycoprotein endopeptidase, indicating that it belongs to the class of sialomucin-like proteins. Therefore, we suggest the name endomucin. Treatment of isolated endomucin by sialidase and O-glycosidase reduced the apparent molecular weight to 45 kD and abolished binding of all three antibodies, indicating that carbohydrates are directly or indirectly involved in the formation of the antibody epitopes. Immunohistological analysis of all examined mouse tissues showed that endomucin is an endothelial antigen found in venous endothelium as well as in capillaries, but not on arterial endothelium. Interestingly, high endothelial venules of peripheral and mesenteric lymph nodes as well as of Peyers's patches were negative for staining with the three MoAbs. PMID- 9864159 TI - Mild hemophilia A caused by increased rate of factor VIII A2 subunit dissociation: evidence for nonproteolytic inactivation of factor VIIIa in vivo. AB - Approximately 5% of hemophilia A patients have normal amounts of a dysfunctional factor VIII (FVIII) protein and are termed cross-reacting material (CRM) positive. FVIII is a heterodimer (domain structure A1-A2-B/A3-C1-C2) that requires thrombin cleavage to elicit procoagulant activity. Thrombin-activated FVIII is a heterotrimer with the A2 subunit (amino acid residues 373 to 740) in a weak ionic interaction with the A1 and A3-C1-C2 subunits. Dissociation of the A2 subunit correlates with inactivation of FVIII. Recently, a phenotype of CRM positive hemophilia A patients has been characterized whose plasma displays a discrepancy between their FVIII activities, where the one-stage clotting assay displays greater activity than the two-stage clotting assay. One example is a missense mutation where ARG531 has been substituted by HIS531. An FVIII cDNA construct was prepared containing the ARG531(HIS) mutation and the protein was expressed in COS-1 monkey cells by transient DNA transfection. Metabolic labeling with [35S]-methionine demonstrated that ARG531(HIS) was synthesized at an equal rate compared with FVIII wild-type (WT) but had slightly reduced antigen in the conditioned medium, suggesting a modest secretion defect. A time course of structural cleavage of ARG531(HIS) demonstrated identical thrombin cleavage sites and rates of proteolysis as FVIII WT. Similar to the patient phenotypes, ARG531(HIS) had discrepant activity as measured by a one-stage activated partial thromboplastin time (aPTT) clotting assay (36% +/- 9.6% of FVIII WT) and a variation of the two-stage assay using a chromogenic substrate (COAMATIC; 19% +/- 6.9% of FVIII WT). Partially purified FVIII WT and ARG531(HIS) proteins were subjected to functional activation by incubation with thrombin. ARG531(HIS) demonstrated significantly reduced peak activity and was completely inactivated after 30 seconds, whereas FVIII WT retained activity until 2.5 minutes after activation. Because the ARG531(HIS) missense mutation predicts a charge change to the A2 subunit, we hypothesized that the ARG531(HIS) A2 subunit could be subject to more rapid dissociation from the heterotrimer. The rate of A2 dissociation, using an optical biosensor, was determined to be fourfold faster for ARG531(HIS) compared with FVIII WT. Because the two-stage assay involves a preincubation phase before assay measurement, an increased rate of A2 dissociation would result in an increased rate of inactivation and reduced specific activity. PMID- 9864160 TI - The vascular endothelial-cadherin promoter directs endothelial-specific expression in transgenic mice. AB - Vascular endothelial-cadherin (VE-cadherin) is a calcium-dependent adhesive molecule, exclusively and constitutively expressed in endothelial cells. Analysis of the VE-cadherin promoter fused to a reporter gene in bovine aortic endothelial cells showed three major functional regions. The proximal region alone (-139, +24) promoted nonspecific transcription; the addition of the (-289, -140) and ( 2226, -1190) domains abolished transcription in fibroblasts while expression in endothelial cells remained unchanged, suggesting that fragments (-2226, +24) and longer contain the full endogenous promoter activity. To study the transcriptional specificity of the promoter region in vivo, we generated transgenic mice carrying the chimeric construct containing the (-2486, +24) region. The promoter directed reporter expression in all examined organs of adult transgenic mice. During embryonic development, transgene expression was detected at the early steps of vasculogenesis. Later, the expression persisted during development of the vascular system and was restricted to the endothelial layer of the vessels. Together, these data provide evidence for specific regulatory regions within the VE-cadherin promoter. Furthermore, the identification of DNA sequences restricting gene expression to the endothelium has many potential applications for the development of animal models of cardiovascular or angiogenic diseases or for the delivery of therapeutic molecules. PMID- 9864161 TI - von Willebrand factor elevates plasma factor VIII without induction of factor VIII messenger RNA in the liver. AB - Factor VIII and von Willebrand factor (vWF) circulate in the plasma as a noncovalent protein complex. Circulating levels of factor VIII are coordinately regulated with circulating levels of vWF in which the ratio is maintained at 1 molecule of factor VIII for 50 to 100 vWF subunits. Infusion of vWF into vWF deficient animal models and human patients yields a secondary increase in circulating levels of factor VIII. We have studied the mechanism of the secondary rise in factor VIII in a porcine model of vWF deficiency. On infusion of vWF into a vWF-deficient pig there was an approximately fivefold increase in circulating factor VIII activity. Liver biopsies were taken pre- and post-vWF infusion for isolation of total messenger RNA (mRNA). Factor VIII-specific mRNA was measured by an RNAse protection assay. The results showed no difference in the liver specific factor VIII mRNA on vWF infusion. These results indicate that the secondary rise in factor VIII levels in response to exogenous vWF infusion is not dependent on increased steady-state levels of factor VIII mRNA in the liver. PMID- 9864162 TI - Light chain shifting: identification of a human plasma cell line actively undergoing light chain replacement. AB - We identified an antibody-secreting human B-cell line (HTD8), which actively replaces the production of the original lambda light chain with a new lambda chain (light chain shifting) at a high rate. Loss of the original rearranged lambda light chain occurs by significantly reducing the amount of transcript expressed. Expression of the new lambda chain, which replaces the original lambda chain, occurs by rearranging new VJ segments on a previously excluded allele. V lambda gene usage of these new rearrangements are biased toward Vlambda4, Vlambda6, and Vlambda10 families, which are known to be the least frequently used. In striking contrast to the plasma cell phenotype, recombination activating genes, RAG-1 and RAG-2, were expressed in the HTD8 cells and were shown to be necessary, but insufficient for inducing expression of the new lambda chain. These results suggest that human plasma cells have the potential to actively undergo light chain replacement. PMID- 9864163 TI - Mechanism of interleukin-10 inhibition of T-helper cell activation by superantigen at the level of the cell cycle. AB - We have analyzed the effects of interleukin-10 (IL-10) on the entry of quiescent CD4(+) T cells into the cell cycle upon stimulation with the superantigen staphylococcal enterotoxin B (SEB). IL-10 arrested cells at G0/G1. IL-10 treatment prevented the downregulation of p27(Kip1), an inhibitory protein that controls progression out of the G0 phase of the cell cycle. IL-10 also prevented the upregulation of the G1 cyclins D2 and D3, proteins necessary for entry and progression through the G1 phase of the cell cycle. Associated with the inhibition of the cell cycle, IL-10 suppressed SEB induction of interleukin-2 (IL 2). Addition of exogenous IL-2 to IL-10-treated cells significantly reversed the antiproliferative effects of IL-10. Moreover, IL-10 effects on the early G1 proteins p27(Kip1) and cyclin D2 were similarly reversed by exogenous IL-2. Although this reversal by IL-2 was pronounced, it was not complete, suggesting that IL-10 may have some effects not directly related to the suppression of IL-2 production. Cell separation experiments suggest that IL-10 can effect purified CD4(+) T cells directly, providing functional evidence for the presence of IL-10 receptors on CD4(+) T cells. IL-10 also inhibited expression of IL-2 transcriptional regulators c-fos and c-jun, which also inhibit other cell functions. Our studies show that the mechanism of IL-10 regulation of quiescent CD4(+) T-cell activation is mainly by blocking induction of IL-2 that is critical to downregulation of p27(Kip1) and upregulation of D cyclins in T-cell activation and entry into the cell cycle. PMID- 9864164 TI - Ceramide and cyclic adenosine monophosphate (cAMP) induce cAMP response element binding protein phosphorylation via distinct signaling pathways while having opposite effects on myeloid cell survival. AB - The role of ceramide as a second messenger is a subject of great interest, particularly since it is implicated in signaling in response to inflammatory cytokines. Ceramide induces apoptosis in both cytokine-dependent MC/9 cells and factor-independent U937 cells. Elevation of cyclic adenosine monophosphate (cAMP) levels inhibits apoptosis induced by ceramide and several other treatments. One target of cAMP-mediated signaling is the transcription factor CREB (cAMP response element binding protein), and recently CREB phosphorylation at an activating site has been shown to also be mediated by a cascade involving p38 mitogen-activated protein kinase (MAPK), one of the stress-activated MAP kinases. Because no role for p38 MAPK in apoptosis has been firmly established, we examined the relationship between p38 MAPK and CREB phosphorylation under various conditions. Ceramide, or sphingomyelinase, like tumor necrosis factor- (TNF-) or the hematopoietic growth factor, interleukin-3 (IL-3), was shown to activate p38 MAPK, which in turn activated MAPKAP kinase-2. Each of these treatments led to phosphorylation of CREB (and the related factor ATF-1). A selective p38 MAPK inhibitor, SB203580, blocked TNF-- or ceramide-induced CREB phosphorylation, but had no effect on the induction of apoptosis mediated by these agents. The protective agents cAMP and IL-3 also led to CREB phosphorylation, but this effect was independent of p38 MAPK, even though IL-3 was shown to activate both p38 MAPK and MAPKAP kinase-2. Therefore, the opposing effects on apoptosis observed with cAMP and IL-3, compared with ceramide and TNF-, could not be explained on the basis of phosphorylation of CREB. In addition, because SB203580 had no effect of TNF- or ceramide-induced apoptosis, our results strongly argue against a role for p38 MAPK in the induction of TNF-- or ceramide-induced apoptosis. PMID- 9864166 TI - Insulin-like growth factor-1 acts as a chemoattractant factor for 5T2 multiple myeloma cells. AB - The chemotactic and growth-stimulatory effect of insulin-like growth factor 1 (IGF-1) was investigated in the experimental mouse 5T2 multiple myeloma (MM) model. Chemotaxis was analyzed by classical checkerboard analysis. Bone marrow fibroblasts-conditioned medium exhibited a chemotactic effect on 5T2 MM cells that could be neutralized by adding a blocking antibody to IGF-1. On the other hand, exogenously added IGF-1 also had a chemotactic effect on the 5T2 MM cells. Moreover, in vitro analysis demonstrated that transmigrated 5T2 MM cells have a higher expression of IGF-1 receptor, both in bone marrow-conditioned medium and in IGF-1-induced chemotaxis, in comparison to cells before migration. When analyzed in vivo, 18 hours after injection of the heterogeneous 5T2 MM population, 5T2 MM cells present in the bone marrow show a higher expression of the IGF-1 receptor than their counterparts before injection. When the proliferative effect of IGF-1 was analyzed, no stimulation was observed, which is in contrast to the influence of bone marrow-conditioned medium and interleukin-6. Our results suggest a causal relationship between the presence of IGF-1 in the bone marrow and the chemotaxis of MM cells to and their subsequent presence in the bone marrow. PMID- 9864165 TI - Marginal-zone B cells in the human lymph node and spleen show somatic hypermutations and display clonal expansion. AB - Splenic marginal-zone B cells, marginal-zone B cells of Peyer's patches in the gut, and nodal marginal-zone B cells (also identified as monocytoid B cells) share a similar morphology and immunophenotype. These cells likely represent a distinct subset of B cells in humans and rodents, but their precise ontogenetic relationship as well as their origin from B cells of the germinal center is still debated. To study this, we performed a mutation analysis of the rearranged immunoglobulin variable genes (VH) of microdissected single nodal and splenic marginal-zone cells. In addition, we investigated the presence of proliferating cells and B-cell clones in the human splenic and nodal marginal zone as well as adjacent germinal centers. This was performed by immunohistochemical staining for the Ki-67 antigen and denaturing gradient gel analysis of amplified immunoglobulin heavy chain genes' complementarity determining region 3 of microdissected cell clusters. A variable subset of nodal and splenic marginal zone B cells showed somatic mutations in their rearranged VH genes, indicating that both virgin and memory B cells are present in the nodal and splenic marginal zone. Nodal and splenic marginal-zone B cells preferentially rearranged VH3 family genes such as DP47, DP49, DP54, and DP58. A preferential rearrangement of the same VH genes has been shown by others in the peripheral CD5(-) IgM+ B cells. These data suggest that the splenic and nodal marginal-zone B cells are closely related B-cell subsets. We also showed that marginal-zone B cells may cycle and that clones of B cells are frequently detected in the nodal as well as the splenic marginal zone. These clones are not related to those present in adjacent germinal centers. These data favor the hypothesis that clonal expansion occurs in the marginal zone. Whether the somatic hypermutation mechanism is activated during the clonal expansion in the marginal zone and which type of immune response triggers the clonal expansion need to be elucidated. PMID- 9864167 TI - T-cell receptor analysis in Omenn's syndrome: evidence for defects in gene rearrangement and assembly. AB - Patients with Omenn's syndrome have a form of severe immune deficiency that is associated with pathological features of graft-versus-host disease, except for the lack of foreign engraftment. It has been hypothesized that the disease's unique clinical features are mediated by an expanded population of autologous self-reactive T cells of limited clonality. In the current study, an investigation of the T-cell receptor (TCR) repertoire was undertaken to identify defects in T-cell rearrangement and development. The TCR repertoire in this group of patients was exquisitely restricted in the number of different TCR clonotypes, and some of these clonotypes seemed to have similar recognition motifs in the antigen-binding region, indicating antigen-driven proliferation of T lymphocytes. The TCRs from some patients lacked N- or P-nucleotide insertions and used proximal variable and joining gene segments, suggesting abnormal intrathymic T cell development. Finally, abnormal assembly of gene segments and truncated rearrangements within nonproductive alleles suggested abnormalities in TCR rearrangement mechanisms. Overall, the findings suggest that inefficient and/or abnormal generation of TCRs may be a consistent feature of this disease. PMID- 9864168 TI - Hairy cell leukemia-specific recognition by multiple autologous HLA-DQ or DP restricted T-cell clones. AB - We studied in patients with hairy cell leukemia (HCL) whether autoreactive T cells could be isolated with specific reactivity to the HCL cells. HCL cells were activated via triggering of CD40 on the cell membrane and used as stimulator cells to generate autologous T-cell clones. Two types of CD4(+)BV2(+) T-cell clones with different CDR3 rearrangements and one type of CD4(+)BV8S3(+) T-cell clone were generated from the spleen or blood. These clones specifically recognized the autologous HCL cells, without reactivity to autologous peripheral blood mononuclear cells (PBMC), phytohemagglutinin blasts, or Epstein-Barr virus transformed B cells in a primed lymphocyte test. Blocking and panel studies using HCL cells from 11 other patients showed that recognition of the HCL cells by the BV2(+) T cells was restricted by HLA-DQA1*03/DQB1*0301, and the BV8S3(+) T cells were restricted by DPB1*04. The T-cell clones did not recognize DPB1*04(+) or DQ3(+) PBMC from healthy donors or DP/DQ matched malignant cells from patients with other hematologic malignancies, except for one patient with acute lymphoblastic leukemia. These HCL-specific T-cell clones may be used for the detection of an HCL-specific tumor antigen. PMID- 9864169 TI - Constitutive and interleukin-7/interleukin-15 stimulated DNA binding of Myc, Jun, and novel Myc-like proteins in cutaneous T-cell lymphoma cells. AB - Members of the Myc and Jun/Fos gene families have been found to be expressed in late stages of cutaneous T-cell lymphoma (CTCL) and may be responsible for the transition from low-grade to high-grade tumors. The composition of these complexes is an important parameter, as the different homo- and heterodimeric jun and myc complexes can have gene transcription activating or suppressing activities. We determined the composition of the jun and myc DNA-binding complexes in three CTCL cell lines and malignant cells of seven Sezary patients by electrophoretic mobility shift assays (EMSAs) and "supershift" assays in which specific antibodies against the different members of the tested gene families were included in the binding reactions. Complexes containing JunD were found in three cell lines and two patients. The three cell lines and one patient contained also c-Myc/Max heterodimers. Because c-Myc/Max heterodimers are strong gene transcription activators and are necessary for cell-cycle progression, they may play a role in the progression of CTCL. JunD may also promote cell-cycle progression and influence the expression of cell death survival genes. Interleukin-7 (IL-7) and IL-15, which have been identified as growth factors for CTCL cells, stimulated the DNA binding of JunD and two novel c-Myc recognition site (E-box) binding proteins, but not the DNA binding of c-Myc/Max heterodimers. PMID- 9864170 TI - Malignant cells can be sensitized to undergo growth inhibition and apoptosis by arsenic trioxide through modulation of the glutathione redox system. AB - Arsenic trioxide (As2O3) induces clinical remission in acute promyelocytic leukemia (APL) with minimal toxicity and apoptosis in APL-derived NB4 cells at low (1 to 2 micromol/L) concentration. We examined the basis for NB4 cell sensitivity to As2O3 to identify experimental conditions that would render other malignant cells responsive to low concentrations of As2O3. The intracellular glutathione (GSH) content had a decisive effect on As2O3-induced apoptosis. Highly sensitive NB4 cells had the lowest GSH and the sensitivity of other cell lines was inversely proportional to their GSH content. The t(14;18) B-cell lymphoma cell line had low GSH levels and sensitivity to As2O3 at levels slightly higher than in APL cells. Experimental upmodulation of GSH content decreased the sensitivity to As2O3. Ascorbic acid and buthionine sulfoxide (BSO) decreased GSH to a greater extent, and rendered malignant cells more sensitive to As2O3. As2O3 induced apoptosis was not enhanced by ascorbic acid in normal cells, suggesting that the combination of ascorbic acid and As2O3 may be selectively toxic to some malignant cells. Ascorbic acid enhanced the antilymphoma effect of As2O3 in vivo without additional toxicity. Thus, As2O3 alone or administered with ascorbic acid may provide a novel therapy for lymphoma. PMID- 9864171 TI - Arsenic trioxide and interferon-alpha synergize to induce cell cycle arrest and apoptosis in human T-cell lymphotropic virus type I-transformed cells. AB - Human T-cell lymphotropic virus type I (HTLV-I) is the causative agent of adult T cell leukemia/lymphoma (ATL). ATL is an aggressive proliferation of mature activated T cells associated with a poor prognosis. The combination of the antiviral agents, zidovudine (AZT) and interferon (IFN), is a potent treatment of ATL. Recently, arsenic trioxide (As) was shown to be an effective treatment of acute promyelocytic leukemia (APL). We have tested the effects of the combination of As and IFN on cell proliferation, cell cycle phases distribution, and apoptosis in ATL-derived or control T-cell lines. A high synergistic effect between IFN and As was observed in ATL-derived cell lines in comparison to the control cell lines, with a dramatic inhibition of cell proliferation, G1 arrest, and induction of apoptosis. Similar results were obtained with fresh leukemia cells derived from an ATL patient. Although the mechanisms involved are unclear, these results could provide a rational basis for combined As and IFN treatments in ATL. PMID- 9864172 TI - BCR/ABL- CD34(+)HLA-DR- progenitor cells in early chronic phase, but not in more advanced phases, of chronic myelogenous leukemia are polyclonal. AB - Chronic myelogenous leukemia (CML) is characterized by the Philadelphia (Ph) translocation and BCR/ABL gene rearrangement which occur in a pluripotent hematopoietic progenitor cell. Ph-negative (Ph-) hematopoiesis can be restored in vivo after treatment with -interferon or intensive chemotherapy, suggesting that normal stem and progenitor cells coexist with the Ph+ clone. We have previously shown that Ph- progenitors are highly enriched in the CD34(+)HLA-DR- fraction from early chronic phase (ECP) CML patients. Previous studies have suggested that the Ph-translocation represents a secondary clonal hit occurring in an already clonally mutated Ph- progenitor or stem cells, leaving the unanswered question whether Ph- CD34(+)HLA-DR- progenitors are normal. To show the clonal nature of Ph- CD34(+)HLA-DR- CML progenitors, we have compared the expression of BCR/ABL mRNA with X-chromosome inactivation patterns (HUMARA) in mononuclear cells and in CD34(+)HLA-DR+ and CD34(+)HLA-DR- progenitors in marrow and blood obtained from 11 female CML patients (8 in chronic phase and 3 in accelerated phase [AP] disease). Steady-state marrow-derived BCR/ABL mRNA-, CD34(+)HLA-DR- progenitors had polyclonal X-chromosome inactivation patterns in 2 of 2 patients. The same polyclonal pattern was found in the progeny of CD34(+)HLA-DR- derived long-term culture-initiating cells. Mobilization with intensive chemotherapy induced a Ph-, BCR/ABL mRNA- and polyclonal state in the CD34(+)HLA-DR- and CD34(+)HLA-DR+ progenitors from 2 ECP patients. In a third ECP patient, polyclonal CD34(+) cells could only be found in the first peripheral blood collection. In contrast to ECP CML, steady-state marrow progenitors in late chronic phase and AP disease were mostly Ph+, BCR/ABL mRNA+, and clonal. Further, in the majority of these patients, a Ph-, polyclonal state could not be restored despite mobilization with intensive chemotherapy. We conclude from these studies that CD34(+)HLA-DR- cells that are Ph- and BCR/ABL mRNA- are polyclonal and therefore benign. This population is suitable for autografting in CML. PMID- 9864173 TI - ETV6-AML1 translocation breakpoints cluster near a purine/pyrimidine repeat region in the ETV6 gene. AB - The t(12;21)(p13;q22) translocation, fusing the ETV6 and AML1 genes, is the most frequent chromosomal translocation associated with pediatric B-cell precursor acute lymphoblastic leukemia. Although the genomic organization of the ETV6 gene and a breakpoint cluster region (bcr) in ETV6 intron 5 has been described, mapping of AML1 breakpoints has been hampered because of the large, hitherto unknown size of AML1 intron 1. Here, we report the mapping of the AML1 gene between exons 1 and 3, cloning of ETV6-AML1 breakpoints from different patients, and localization of the AML1 breakpoints within AML1 intron 1. In contrast to the tightly clustered ETV6 breakpoints, the AML1 breakpoints were found to be dispersed throughout AML1 intron 1. Although nucleotide sequence analysis of the breakpoint junctions showed several 5/7 matches for the V(D)J consensus heptamer recognition sequence, these matches were present only on the ETV6 alleles and not on the AML1 alleles, making it unlikely that the translocations were mediated by a simple V(D)J recombination mistake. Interestingly, several breakpoints as well as a stable insertion polymorphism mapped close to a polymorphic, alternating purine-pyrimidine tract in the ETV6 gene, suggesting that this region may be prone to DNA recombination events such as insertions or translocations. Finally, the presence of an insertional polymorphism within the ETV6 bcr must be recognized to avoid incorrect genotype designation based on Southern blot analysis. PMID- 9864174 TI - Comparison of interleukin-1 beta expression by in situ hybridization in monoclonal gammopathy of undetermined significance and multiple myeloma. AB - We investigated whether interleukin-1beta (IL-1beta) is differentially expressed in plasma cells from monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM) patients because IL-1beta appears to play a major role in the development of lytic bone lesions, the major clinical feature distinguishing MGUS from myeloma. In situ hybridization (ISH) for IL-1beta was performed using bone marrow aspirates from 51 MM, 7 smoldering MM, 21 MGUS, and 5 normal control samples. Using the ISH technique IL-1beta mRNA was detectable in the plasma cells from 49 of 51 patients with active myeloma and 7 of 7 patients with smoldering myeloma. In contrast, 5 of 21 patients with MGUS and 0 of 5 normal controls had detectable IL-1beta message. Bone lesions were present in 40 of the 51 MM patients analyzed, and all 40 patients had IL-1beta mRNA by ISH. These results show that greater than 95% of MM patients but less than 25% of MGUS patients are positive for IL-1beta production. In the future, continued follow-up of IL-1beta positive and negative MGUS patients should determine whether aberrant expression of plasma cell IL-1beta is predictive of those MGUS patients that will eventually progress to active myeloma. PMID- 9864175 TI - Efflux of rhodamine from CD56+ cells as a surrogate marker for reversal of P glycoprotein-mediated drug efflux by PSC 833. AB - The expression of high levels of P-glycoprotein (Pgp) in circulating mononuclear cells allowed us to use an ex vivo assay as a surrogate measure of Pgp antagonism. Efflux of rhodamine from CD56(+) cells was measured before the start of PSC 833 and at varying times thereafter. Patients receiving PSC 833 had decreased rhodamine efflux from their circulating CD56(+) cells. Time course studies showed that following a single oral dose of PSC 833, decreased rhodamine efflux was found in some patients within 15 minutes of treatment. Maximal inhibition was observed at times ranging from 45 minutes to 60 minutes. A dose response relationship was shown between the concentration of PSC 833 in the blood and the inhibition of rhodamine efflux, with an apparent plateau of the inhibition of rhodamine efflux at approximately 1,000 ng/mL. The Ki, defined as the concentration required for half-maximal inhibition of Pgp-mediated rhodamine efflux, was determined to be in the range of 29 to 181 ng/mL; although results in two patients were distinctly different, with Ki values of 914 and 916 ng/mL. MRK 16 staining was similar among all patients. We conclude that measurement of rhodamine efflux from CD56(+) cells provides a surrogate assay with the potential for monitoring Pgp antagonism in clinical trials. PMID- 9864176 TI - Hyperdiploid acute lymphoblastic leukemia with 51 to 65 chromosomes: a distinct biological entity with a marked propensity to undergo apoptosis. AB - To determine the cellular basis for the excellent clinical outcome of hyperdiploid acute lymphoblastic leukemia (ALL), defined by a modal chromosome number of 51 to 65, we assessed the growth potential of leukemic cells from 129 children with newly diagnosed ALL. Flow cytometric analysis was used to compare leukemic cell recoveries at the beginning and at the end of 7-day cultures on allogeneic bone marrow-derived stromal layers. The median percentage of cell recovery after culture was 91% (range, <1% to 550%). Among the 25 hyperdiploid cases, only two had cell recoveries above the median value, compared with 63 of 104 cases with different ploidies (P <.001); 21 had recoveries within the first quartile, in contrast to only 12 of the 104 other cases. Cell recoveries in the 16 cases with duplications of chromosomes 4 and 10, a feature previously associated with a superior outcome, were all within the first quartile. Flow cytometric studies indicated that rapid induction of apoptosis was the underlying cause of low cell recoveries in cases with hyperdiploidy. The demise of hyperdiploid cells on stroma was not due to failure to adhere with stromal elements (as shown by electron microscopy) or to deficiencies of interleukin-1 (IL-1), IL-2, IL-3, IL-4, IL-6, IL-7, IL-11, stem-cell factor, interferon- (IFN ), tumor necrosis factor- (TNF-), or to combinations of these cytokines. Inactivation of IL-4, IFN- and TNF-, which if secreted by stromal layers could be toxic to ALL cells, failed to improve the survival of hyperdiploid blasts. We conclude that leukemic cells bearing 51 to 65 chromosomes have a marked propensity to undergo apoptosis. The stringent survival requirements of these cells, together with their potentially higher sensitivity to antileukemic drugs, may well account for the high cure rates achieved in patients with this form of ALL. PMID- 9864177 TI - Two candidate downstream target genes for E2A-HLF. AB - The E2A-HLF fusion gene, formed by the t(17;19)(q22;p13) chromosomal translocation, is thought to drive the leukemic transformation of early B-cell precursors by repressing an evolutionarily conserved apoptotic pathway. To test this hypothesis, we sought to identify downstream targets of E2A-HLF in t(17;19)+ pro-B leukemia cells (UOC-B1) that had been transfected with a zinc-inducible vector encoding a dominant-negative suppressor (E2A-HLF[dn]) of the oncoprotein. Representational difference analysis of mRNAs from E2A-HLF(dn)+ UOC-B1 cells grown with (E2A-HLF inactive) or without (E2A-HLF active) the addition of zinc yielded several differentially expressed cDNA fragments that were individually subcloned. Two of the clones, designated F-5 and G-4, hybridized with mRNAs that were upregulated by E2A-HLF. Levels of both transcripts declined sharply within 8 to 12 hours after suppression of E2A-HLF DNA-binding activity, becoming undetectable after 96 hours. The F-5 cDNA was identified as a portion of ANNEXIN VIII, whose product was expressed in promyelocytic leukemia cells and UOC-B1 cells, but not in other leukemic cell lines. A novel full-length cDNA cloned with the G-4 fragment encoded a protein that we have named SRPUL (sushi-repeat protein upregulated in leukemia). It is normally expressed in heart, ovary, and placenta, but could not be detected in leukemic cell lines other than UOC-B1. Neither protein prevented apoptosis in interleukin-3-dependent murine pro-B cells, suggesting that they have paraneoplastic roles in leukemias that express E2A-HLF, perhaps in the disseminated intravascular coagulopathy and hypercalcemia that characterize these cases. PMID- 9864178 TI - Nitric oxide-mediated augmentation of polymorphonuclear free radical generation after hypoxia-reoxygenation. AB - Polymorphonuclear leukocytes (PMNLs), nitric oxide (NO), calcium, and free radicals play an important role in hypoxia/ischemia and reoxygenation injury. In the present study, NO donors, sodium nitroprusside (SNP), and diethylamine-NO (DEA-NO) at low concentrations (10 and 100 nmol/L) potentiated, while higher (10 micromol/L to 10 mmol/L) concentrations inhibited free radical generation response in the rat PMNLs. Free radical generation response was found to be significantly augmented when hypoxic PMNLs were reoxygenated (hypoxia reoxygenation [H-R]). This increase in free radical generation after reoxygenation or SNP (10 nmol/L) was blocked in the absence of extracellular calcium. SNP (10 nmol/L) or H-R-mediated increases in the free radical generation were prevented by the pretreatment of PMNLs with NO scavenger (hemoglobin), the polyadenine diphosphate (ADP)-ribosylation synthase inhibitor (benzamide) or the calcium channel antagonist (felodipine). A significant augmentation in the nitrite and intracellular calcium levels was observed during hypoxia. Hemoglobin pretreatment also blocked the increase in intracellular calcium levels due to SNP (10 nmol/L) or hypoxia. Thus, increased availability of NO during SNP treatment or H-R, may have led to an ADP-ribosylation-mediated increase in intracellular calcium, thereby increasing the free radical generation from the rat PMNLs. PMID- 9864179 TI - Cytokine-specific activation of distinct mitogen-activated protein kinase subtype cascades in human neutrophils stimulated by granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor-alpha. AB - To clarify the differences of the signaling pathways used by granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM CSF), and tumor necrosis factor- (TNF), we investigated activation of mitogen activated protein kinase (MAPK) subtype cascades in human neutrophils stimulated by these cytokines. G-CSF exclusively tyrosine-phosphorylated extracellular signal-regulated kinase (ERK). GM-CSF tyrosine-phosphorylated ERK strongly and p38 MAPK weakly, whereas TNF tyrosine-phosphorylated p38 MAPK strongly and ERK weakly. Consistent with these findings, MEK, an upstream kinase of ERK, was phosphorylated by G-CSF, GM-CSF, and TNF, whereas MKK3/MKK6, an upstream kinase of p38 MAPK, was phosphorylated by GM-CSF and TNF, but not by G-CSF. The potency of these cytokines to phosphorylate ERK and MEK was GM-CSF > G-CSF > TNF, whereas that to phosphorylate p38 MAPK and MKK3/MKK6 was TNF > GM-CSF. C-Jun amino terminal kinase (JNK) was not tyrosine-phosphorylated by any cytokine despite the existence of JNK proteins in human neutrophils, whereas it was tyrosine phosphorylated by TNF in undifferentiated and all-trans retinoic acid differentiated HL-60 cells. Increased phosphorylation of ERK or p38 MAPK was detected within 1 to 5 minutes after stimulation with each cytokine and was dependent on the concentrations of cytokines used. MEK inhibitor (PD98059) reduced tyrosine phosphorylation of ERK, but not p38 MAPK, induced by G-CSF, GM CSF, or TNF. GM-CSF- or TNF-induced superoxide (O2-) release was inhibited by p38 MAPK inhibitor (SB203580) in a dose-dependent manner, suggesting the possible involvement of p38 MAPK in GM-CSF- or TNF-induced O2- release. The results indicate that G-CSF, GM-CSF, and TNF activate the overlapping but distinct MAPK subtype cascades in human neutrophils and suggest that the differential activation of ERK and p38 MAPK cascades may explain the differences of the effects of these cytokines on human neutrophil functions. PMID- 9864180 TI - Activation of the human immunodeficiency virus-1 long terminal repeat by respiratory burst oxidants of neutrophils. AB - The human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) introduced in association with the luciferase reporter gene into Jurkat T cells was strongly activated by a combination of human neutrophils and phorbol myristate acetate (PMA). Activation was not observed when normal neutrophils were replaced by neutrophils which lack a respiratory burst, ie, from a patient with chronic granulomatous disease (CGD), was strongly inhibited by catalase, was potentiated by vanadate, was stimulated by relatively low concentrations of azide, and was inhibited by selective inhibitors of protein kinase C (PKC). The PMA affected activation in three ways: (1) by directly activating the LTR in Jurkat LTRluc; (2) by inducing a respiratory burst in neutrophils with the formation of H2O2; and (3) by increasing the sensitivity of Jurkat LTRluc to the activating effect of H2O2. When PMA was replaced by opsonized zymosan as the neutrophil stimulus, activation of the LTR was low unless azide was added. Activation in the presence of azide was not seen when CGD neutrophils were used or when catalase was added, suggesting that azide acts by inhibiting the degradation of H2O2. These findings indicate that activation of the HIV-1 LTR in Jurkat T cells can be induced by H2O2 released by neutrophils, particularly when PKC is concomitantly activated. PMID- 9864181 TI - The use of allele-specific recombinant Fc gamma receptor IIIb antigens for the detection of granulocyte antibodies. AB - The Fcgamma receptor IIIb (FcgammaRIIIb) for the Fc domain of IgG is expressed exclusively on neutrophils. The FcgammaRIIIb bears allotypic polymorphisms referred to as NA1, NA2, and SH, which are known for their frequent involvement in alloimmune and autoimmune neutropenias as well as in transfusion reactions. The bactericidal capacity of isolated neutrophils is easily activatable, and activation results in self-desintegration, thus preventing storage of neutrophils. As a result, only freshly isolated granulocytes can be used for antibody screening, often making it impossible to use typed panel cells. To provide a readily available source of typed panel cells, we therefore established stable mammalian cells expressing recombinant NA1, NA2, and SH antigens. We isolated mRNA from typed neutrophils and then transcribed it in cDNA. The cDNA that codes for the different forms of the FcgammaRIIIb was amplified by polymerase chain reaction and was subsequently subcloned into the mammalian expression vector pcDNA3. Chinese hamster ovary (CHO) cells were transfected with allele-specific constructs, and stable cell lines expressing FcgammaRIIIb were selected by flow cytometry. Because human sera show high background fluorescence with transfectants in flow cytometry, the monoclonal antibody-specific isolation of granulocyte antigens (MAIGA) assay was performed. By MAIGA assay, we tested 14 well-characterized human alloantibodies directed against the antigens NA1, NA2, and SH; 5 FcgammaRIIIb-specific isoantibodies; and 12 FcgammaRIIIb-reactive autoantibodies. Except one NA1- and one SH-specific alloantibody, all other antibodies could be identified by the use of CHO transfectants. In contrast to neutrophils, fixed CHO cells can be stored at 4 degrees C for at least 4 weeks or stored frozen for a longer period. This longer shelf life of the transfected CHO cells compared with isolated neutrophils will simplify the detection of the clinically most important FcgammaRIIIb-reactive alloantibodies and autoantibodies. PMID- 9864182 TI - Protein replacement by receptor-mediated endocytosis corrects the sensitivity of Fanconi anemia group C cells to mitomycin C. AB - Current methods for direct gene transfer into hematopoietic cells are inefficient. Here we show that functional complementation of Fanconi anemia (FA) group C cells by protein replacement can be as efficacious as by transfection with wild-type FAC cDNA. We expressed a chimeric protein (called His-ILFAC) consisting of the mature coding portion of gibbon interleukin-3 (IL-3) and full length FAC in Escherichia coli. The purified bacterial protein is internalized by hematopoietic cells via IL-3 receptors. The intracellular half-life of His-ILFAC is approximately 60 minutes, which is comparable to that of the transgene-encoded FAC protein. In this cell-culture model His-ILFAC completely corrects the sensitivity of FA group C cells to mitomycin C, but it has no effect on FA cells that belong to complementation groups A and B. We suggest that receptor-mediated endocytosis of cytokine-fusion proteins may be of general use to deliver macromolecules into hematopoietic progenitor cells. PMID- 9864183 TI - HBED: the continuing development of a potential alternative to deferoxamine for iron-chelating therapy. AB - To further examine the potential clinical usefulness of the hexadentate phenolic aminocarboxylate iron chelator N, N'-bis(2-hydroxybenzyl)ethylenediamine-N,N' diacetic acid (HBED) for the chronic treatment of transfusional iron overload, we performed a subchronic toxicity study of the HBED monosodium salt in rodents and have evaluated the iron excretion in primates induced by HBED. The HBED-induced iron excretion was determined for the monohydrochloride dihydrate that was first dissolved in a 0.1-mmol/L sodium phosphate buffer at pH 7.6 and administered to the primates either orally (PO) at a dose of 324 micromol/kg (149.3 mg/kg, n = 5), subcutaneously (sc) at a dose of 81 micromol/kg (37.3 mg/kg, n = 5), sc at 324 micromol/kg (n = 5), and sc at 162 micromol/kg (74.7 mg/kg) for 2 consecutive days for a total dose of 324 micromol/kg (n = 3). In addition, the monosodium salt of HBED in saline was administered to the monkeys sc at a single dose of 150 micromol/kg (64.9 mg/kg, n = 5) or at a dose of 75 micromol/kg every other day for three doses, for a total dose of 225 micromol/kg (n = 4). For comparative purposes, we have also administered deferoxamine (DFO) PO and sc in aqueous solution at a dose of 300 micromol/kg (200 mg/kg). In the iron-loaded Cebus apella monkey, whereas the PO administration of DFO or HBED even at a dose of 300 to 324 micromol/kg was ineffective, the sc injection of HBED in buffer or its monosodium salt, 75 to 324 micromol/kg, produced a net iron excretion that was nearly three times that observed after similar doses of sc DFO. In patients with transfusional iron overload, sc injections of HBED may provide a much needed alternative to the use of prolonged parenteral infusions of DFO. Note: After the publication of our previous paper (Blood, 91:1446, 1998) and the completion of the studies described here, it was discovered that the HBED obtained from Strem Chemical Co (Newburyport, MA) that was labeled and sold as a dihydrochloride dihydrate was in fact the monohydrochloride dihydrate. Therefore, the actual administered doses were 81, 162, or 324 micromol/kg; not 75, 150, or 300 micromol/kg as was previously reported. The new data have been recalculated accordingly, and the data from our earlier study, corrected where applicable, are shown in parentheses. PMID- 9864184 TI - Evaluation of biochemical changes during in vivo erythrocyte senescence in the dog. AB - One hypothesis to explain the age-dependent clearance of red blood cells (RBCs) from circulation proposes that denatured/oxidized hemoglobin (hemichromes) arising late during an RBC's life span induces clustering of the integral membrane protein, band 3. In turn, band 3 clustering generates an epitope on the senescent cell surface leading to autologous IgG binding and consequent phagocytosis. Because dog RBCs have survival characteristics that closely resemble those of human RBCs (ie, low random RBC loss, approximately 115-day life span), we decided to test several aspects of the above hypothesis in the canine model, where in vivo aged cells of defined age could be evaluated for biochemical changes. For this purpose, dog RBCs were biotinylated in vivo and retrieved for biochemical analysis at various later dates using avidin-coated magnetic beads. Consistent with the above hypothesis, senescent dog RBCs were found to contain measurably elevated membrane-bound (denatured) globin and a sevenfold enhancement of surface-associated autologous IgG. Interestingly, dog RBCs that were allowed to senesce for 115 days in vivo also suffered from compromised intracellular reducing power, containing only 30% of the reduced glutathione found in unfractionated cells. Although the small quantity of cells of age >/=110 days did not allow direct quantitation of band 3 clustering, it was nevertheless possible to exploit single-cell microdeformation methods to evaluate the fraction of band 3 molecules that had lost their normal skeletal linkages and were free to cluster in response to hemichrome binding. Importantly, band 3 in RBCs >/=112 days old was found to be 25% less restrained by skeletal interactions than band 3 in control cells, indicating that the normal linkages between band 3 and the membrane skeleton had been substantially disrupted. Interestingly, the protein 4.1a/protein 4.1b ratio, commonly assumed to reflect RBC age, was found to be maximal in RBCs isolated only 58 days after labeling, implying that while this marker is useful for identifying very young populations of RBCs, it is not a very sensitive marker for canine senescent RBCs. Taken together, these data argue that several of the readily testable elements of the above hypothesis implicating band 3 in human RBC senescence can be validated in an appropriate canine model. PMID- 9864185 TI - Molecular basis of weak D phenotypes. AB - A Rhesus D (RhD) red blood cell phenotype with a weak expression of the D antigen occurs in 0.2% to 1% of whites and is called weak D, formerly Du. Red blood cells of weak D phenotype have a much reduced number of presumably complete D antigens that were repeatedly reported to carry the amino acid sequence of the regular RhD protein. The molecular cause of weak D was unknown. To evaluate the molecular cause of weak D, we devised a method to sequence all 10 RHD exons. Among weak D samples, we found a total of 16 different molecular weak D types plus two alleles characteristic of partial D. The amino acid substitutions of weak D types were located in intracellular and transmembraneous protein segments and clustered in four regions of the protein (amino acid positions 2 to 13, around 149, 179 to 225, and 267 to 397). Based on sequencing, polymerase chain reaction-restriction fragment length polymorphism and polymerase chain reaction using sequence specific priming, none of 161 weak D samples investigated showed a normal RHD exon sequence. We concluded, that in contrast to the current published dogma most, if not all, weak D phenotypes carry altered RhD proteins, suggesting a causal relationship. Our results showed means to specifically detect and to classify weak D. The genotyping of weak D may guide Rhesus negative transfusion policy for such molecular weak D types that were prone to develop anti-D. PMID- 9864186 TI - Peripheral blood CD14(+) cells from healthy subjects carry a circular conformation of latent cytomegalovirus genome. AB - The majority of the human population harbors latent cytomegalovirus. Although CD14(+) peripheral blood mononuclear cells have been implicated as sites of latency, the conformation of the latent viral genome in these cells is unknown. In this study, the conformation of viral genomic DNA was assessed in CD14(+) cells from healthy virus seropositive carriers using an electrophoretic separation on native agarose gels in combination with polymerase chain reaction detection. Here we show that the viral genome migrates as a circular plasmid with a mobility equivalent to a circular 230-kb Shigella flexneri megaplasmid marker. Neither linear nor complex or integrated forms of the viral genome were detected. This report provides further evidence that the CD14(+) cell population is an important site of viral latency in the naturally infected human host. Detection of the viral genome as a circular plasmid during latency suggests that this virus maintains its genome in a manner analogous to other herpesviruses where latent viral genome conformation has been studied. PMID- 9864187 TI - DNA typing for HLA-A and HLA-B identifies disparities between patients and unrelated donors matched by HLA-A and HLA-B serology and HLA-DRB1. AB - High incidences of graft failure and graft-versus-host disease in the recipients of bone marrow transplantations (BMT) from unrelated donors (URD) may reflect the existence of allelic disparities between the patient and the URD despite apparent HLA identity at HLA-A, HLA-B, and HLA-DRB1 loci. To identify the extent and pattern of allelic disparities at HLA-A and HLA-B loci, 128 patients and 484 potential URD were evaluated by DNA typing. DNA typing for HLA-A, HLA-B, and HLA DRB1 was performed at Memorial Sloan Kettering Cancer Center. HLA-A and HLA-B serotyping on URD was provided by the registries. By original typing (serology for HLA-A and HLA-B; DNA typing for DRB1) 187, 164, and 133 URD were 6/6, 5/6, and 4/6 matches, respectively. Following DNA typing, however, only 52.9% of the originally 6/6 matched URD remained 6/6, while 38.5%, 7.5%, and 1.1% were found to be 5/6, 4/6, and 3/6 matches. The level of disparity was higher in the originally 5/6 (P <.01) and 4/6 (P <.01) matched URD. A higher level of disparity was seen for HLA-B as compared to HLA-A. In addition, a serotype related variation was also noticed. For example, 24.1% of HLA-A2 and 60.1% of HLA-B35 seromatched URD were genotypically disparate, but no disparities were seen for HLA-A1 and HLA-B8. A higher percentage of HLA-A (67. 4%) compared with HLA-B (35.4%) serologic homozygous URD remained genotypically homozygous (P =.01). The level of allelic disparity was lower (P <.01 for 6/6; P =.02 for 5/6) if the patient had one of the 15 most common haplotypes (A1B8DR3, A2B7DR15, A3B7DR15, etc) in comparison to the rest of the group. Outcome studies will answer the question whether these disparities are associated with a higher rate of immunological complications seen with URD-BMT. PMID- 9864188 TI - Symbiosis of Aeromonas veronii biovar sobria and Hirudo medicinalis, the medicinal leech: a novel model for digestive tract associations. AB - Hirudo medicinalis, the medicinal leech, is applied postoperatively in modern medicine. Infections by Aeromonas occur in up to 20% of patients unless a preemptive antibiotic treatment is administered. The associated infections demonstrate the need for a better understanding of the digestive tract flora of H. medicinalis. Early studies reported the presence of a single bacterial species in the digestive tract and suggested that these bacteria were endosymbionts contributing to the digestion of blood. In this study, we cultivated bacteria from the digestive tract and characterized them biochemically. The biochemical test results identified the isolates as Aeromonas veronii biovar sobria. This species identification was supported by sequence comparison of a variable region of the genes coding for 16S rRNA. In a colonization assay, a rifampin-resistant derivative of a symbiotic isolate was fed in a blood meal to H. medicinalis. The strain colonized the digestive tract rapidly and reached a concentration similar to that of the native bacterial flora. For the first 12 h, the in vivo doubling time was 1.2 h at 23 degreesC. After 12 h, at a density of 5 x 10(7) CFU/ml, the increase in viable counts ceased, suggesting a dramatic reduction in the bacterial growth rate. Two human fecal isolates, identified as Aeromonas hydrophila and A. veronii biovar sobria, were also able to colonize the digestive tract. These data demonstrate that the main culturable bacterium in the crop of H. medicinalis is A. veronii biovar sobria and that the medicinal leech can be used as a model for digestive tract association of Aeromonas species. PMID- 9864189 TI - Nucleotide sequences of genes coding for fimbrial proteins in a cryptic genospecies of Haemophilus spp. isolated from neonatal and genital tract infections. AB - Nineteen isolates belonging to a cryptic genospecies of Haemophilus (referred to here as genital strains) isolated from genital tract infections (6 strains) and from neonatal infections (13 strains) were studied for fimbrial genes. Sixteen strains exhibit peritrichous fimbriae observed by electron microscopy. By PCR with primers corresponding to the extreme ends of the Haemophilus influenzae type b (Hib) hifA and hifD genes and Southern blotting, a hifA-like gene (named ghfA) and a hifD-like gene (named ghfD) were identified in 6 of the 19 strains. Five of these six strains were from the genital tracts of adults, and one was from a neonate. For each gene, the nucleotide sequence was identical for the six strains. A hifE-like gene (named ghfE) was amplified from only one of the 19 genital strains of Haemophilus, but the ghfE probe gave a signal in Southern hybridization with the five other strains positive for ghfA and ghfD. Therefore, these strains may carry a ghfE-like gene. The Hib fimbrial gene cluster is located between the purE and pepN genes as previously described. For the 13 genital Haemophilus strains that lack fimbrial genes, this region corresponds to a noncoding sequence. Another major fimbrial gene designated the fimbrin gene was previously identified in a nontypeable H. influenzae strain. A fimbrin-like gene was identified for all of our 19 genital strains. This gene is similar to the ompP5 gene of many Haemophilus strains. Therefore, other, unidentified genes may explain the piliation observed in electron microscopy on genital Haemophilus strains which do not possess LKP-like fimbrial genes. Fimbrial genes were significantly associated with strains isolated from the genital tract. They may confer on the strain the ability to survive in the genital tract. PMID- 9864190 TI - Glucose stimulates phagocytosis of unopsonized Pseudomonas aeruginosa by cultivated human alveolar macrophages. AB - Glucose has previously been shown to increase the in vitro phagocytosis of unopsonized Pseudomonas aeruginosa by freshly explanted murine peritoneal macrophages (PM) and cultivated alveolar macrophages (AM). This study examined the effect of glucose on the same phagocytosis process in human AM in order to determine whether this phenomenon is conserved among species. Freshly explanted human AM phagocytosed unopsonized P. aeruginosa at a low level (2 bacteria/macrophage/30 min), whereas mouse AM ingested a negligible number of P. aeruginosa (0.01 bacterium/macrophage/30 min). Glucose had no effect on this or other phagocytic processes in freshly explanted mouse or human AM. However, following in vitro cultivation for 72 h, human AM phagocytosed three to four times more unopsonized P. aeruginosa than did freshly explanted cells, but only in the presence of glucose. This glucose-inducible phagocytic response had also been observed in cultivated murine AM. Although similar increases were also detected for the phagocytosis of latex particles and complement-coated sheep erythrocytes by cultivated human AM, these processes were not glucose dependent. The lack of response to glucose in freshly explanted mouse AM was attributed to insufficient glucose transport; however, freshly explanted human AM exhibited significant facilitative glucose transport activity that was inhibitable by cytochalasin B and phloretin. Taken together, these results suggest that the process of glucose-inducible phagocytosis of unopsonized P. aeruginosa is conserved among macrophages from different species, including humans, and that AM, but not PM, required cultivation for this glucose effect to occur. Glucose transport by AM appears to be necessary but not sufficient for phagocytosis of unopsonized P. aeruginosa. PMID- 9864191 TI - Lipopolysaccharide stimulates butyric acid-induced apoptosis in human peripheral blood mononuclear cells. AB - We previously reported that butyric acid, an extracellular metabolite from periodontopathic bacteria, induced apoptosis in murine thymocytes, splenic T cells, and human Jurkat T cells. In this study, we examined the ability of butyric acid to induce apoptosis in peripheral blood mononuclear cells (PBMC) and the effect of bacterial lipopolysaccharide (LPS) on this apoptosis. Butyric acid significantly inhibited the anti-CD3 monoclonal antibody- and concanavalin A induced proliferative responses in a dose-dependent fashion. This inhibition of PBMC growth by butyric acid depended on apoptosis in vitro. It was characterized by internucleosomal DNA digestion and revealed by gel electrophoresis followed by a colorimetric DNA fragmentation assay to occur in a concentration-dependent fashion. Butyric acid-induced PBMC apoptosis was accompanied by caspase-3 protease activity but not by caspase-1 protease activity. LPS potentiated butyric acid-induced PBMC apoptosis in a dose-dependent manner. Flow-cytometric analysis revealed that LPS increased the proportion of sub-G1 cells and the number of late stage apoptotic cells induced by butyric acid. Annexin V binding experiments with fractionated subpopulations of PBMC in flow cytometory revealed that LPS accelerated the butyric acid-induced CD3(+)-T-cell apoptosis followed by similar levels of both CD4(+)- and CD8(+)-T-cell apoptosis. The addition of LPS to PBMC cultures did not cause DNA fragmentation, suggesting that LPS was unable to induce PBMC apoptosis directly. These data suggest that LPS, in combination with butyric acid, potentiates CD3(+) PBMC T-cell apoptosis and plays a role in the apoptotic depletion of CD4(+) and CD8(+) cells. PMID- 9864192 TI - Influence of outer surface protein A antibody on Borrelia burgdorferi within feeding ticks. AB - Borrelia burgdorferi, the spirochetal agent of Lyme disease, is transmitted by Ixodes ticks. When an infected nymphal tick feeds on a host, the bacteria increase in number within the tick, after which they invade the tick's salivary glands and infect the host. Antibodies directed against outer surface protein A (OspA) of B. burgdorferi kill spirochetes within feeding ticks and block transmission to the host. In the studies presented here, passive antibody transfer experiments were carried out to determine the OspA antibody titer required to block transmission to the rodent host. OspA antibody levels were determined by using a competitive enzyme-linked immunosorbent assay that measured antibody binding to a protective epitope defined by monoclonal antibody C3.78. The C3.78 OspA antibody titer (>213 microgram/ml) required to eradicate spirochetes from feeding ticks was considerably higher than the titer (>6 microgram/ml) required to block transmission to the host. Although spirochetes were not eradicated from ticks at lower antibody levels, the antibodies reduced the number of spirochetes within the feeding ticks and interfered with the ability of spirochetes to induce ospC and invade the salivary glands of the vector. OspA antibodies may directly interfere with the ability of B. burgdorferi to invade the salivary glands of the vector; alternately, OspA antibodies may lower the density of spirochetes within feeding ticks below a critical threshold required for initiating events linked to transmission. PMID- 9864193 TI - Specificity of infection-induced immunity among Borrelia burgdorferi sensu lato species. AB - The specificity of infection-induced immunity in mice infected with cultured or host-adapted Borrelia burgdorferi sensu lato, the agent of Lyme disease, was examined. Sera obtained from mice following infection with high and low doses of cultured B. burgdorferi sensu stricto, transplantation of infected tissue (host adapted spirochetes), or tick-borne inoculation all showed protective activity in passive immunization assays. Infection and disease were similar in mice infected with cultured spirochetes or by transplantation. Thus, the adaptive form of inoculated spirochetes did not influence the immune response during active infection. Mice infected with B. burgdorferi sensu stricto and then cured of infection with an antibiotic during early or late stages of infection were resistant to challenge with high doses of homologous cultured spirochetes for up to 1 year. In contrast, actively immune mice infected with different Borrelia species (B. burgdorferi sensu lato, B. burgdorferi sensu stricto cN40, Borrelia afzelii PKo, and Borrelia garinii PBi) and then treated with an antibiotic were resistant to challenge with cultured homologous but not heterologous spirochetes. Similar results were achieved for actively immune mice challenged by transplantation and by passive immunization with sera from mice infected with each of the Borrelia species and then challenged with cultured spirochetes. Arthritis and carditis in mice that had immunizing infections with B. afzelii and B. garinii and then challenged by transplantation with B. burgdorferi sensu stricto were equivalent in prevalence and severity to those in nonimmune recipient mice. These results indicate that protective immunity and disease modulating immunity that develop during active infection are universal among species related to B. burgdorferi sensu lato but are species specific. PMID- 9864194 TI - High-level expression of Plasmodium vivax apical membrane antigen 1 (AMA-1) in Pichia pastoris: strong immunogenicity in Macaca mulatta immunized with P. vivax AMA-1 and adjuvant SBAS2. AB - The apical membrane antigen 1 (AMA-1) family is a promising family of malaria blood-stage vaccine candidates that have induced protection in rodent and nonhuman primate models of malaria. Correct conformation of the protein appears to be essential for the induction of parasite-inhibitory responses, and these responses appear to be primarily antibody mediated. Here we describe for the first time high-level secreted expression (over 50 mg/liter) of the Plasmodium vivax AMA-1 (PV66/AMA-1) ectodomain by using the methylotrophic yeast Pichia pastoris. To prevent nonnative glycosylation, a conservatively mutagenized PV66/AMA-1 gene (PV66Deltaglyc) lacking N-glycosylation sites was also developed. Expression of the PV66Deltaglyc ectodomain yielded similar levels of a homogeneous product that was nonglycosylated and was readily purified by ion exchange and gel filtration chromatographies. Recombinant PV66Deltaglyc43-487 was reactive with conformation-dependent monoclonal antibodies. With the SBAS2 adjuvant, Pichia-expressed PV66Deltaglyc43-487 was highly immunogenic in five rhesus monkeys, inducing immunoglobulin G enzyme-linked immunosorbent assay titers in excess of 1:200,000. This group of monkeys had a weak trend showing lower cumulative parasite loads following a Plasmodium cynomolgi infection than in the control group. PMID- 9864195 TI - Characterization of emb, a gene encoding the major adhesin of Streptococcus defectivus. AB - Streptococcus defectivus is one of the nutritionally variant streptococci, a class of viridans group streptococci first isolated from patients with endocarditis and otitis media. In previous studies, NVS-47, a clinical isolate of S. defectivus, was shown to bind to the extracellular matrix. A high-molecular weight surface protein was identified and proposed to be responsible for mediating this binding. In the present study, the gene encoding this protein was identified by transposon mutagenesis and characterized. The gene (emb) was found to be larger than 14 kb and was partially sequenced. It encodes a protein containing at least 50 repeats of 77 amino acids predicted to assume an alternating coiled-coil conformation. The domain responsible for extracellular matrix binding was mapped to the N terminus of the protein. From sequence analysis, Emb is proposed to be the prototype of a new family of streptococcal fibrillar proteins. PMID- 9864196 TI - Murine gamma delta T lymphocytes elicited during Plasmodium yoelii infection respond to Plasmodium heat shock proteins. AB - gamma delta T cells accumulate during Plasmodium infections in both murine and human malarias. The biological role of these cells and the antigens that they recognize are not clearly understood, although recent findings indicate that gamma delta T cells in general influence both innate and antigen-specific adaptive host responses. We examined the accumulation of gamma delta T cells elicited during infection with virulent and avirulent Plasmodium yoelii parasites in relatively susceptible and resistant strains of mice. Our results indicated that in nonlethal malaria infections, gamma delta T cells comprise a larger proportion of splenic T cells than in lethal infections and that only a live infection is capable of inducing an increase in the percentage of gamma delta T cells in vivo. Furthermore, we demonstrate that gamma delta T cells elicited during a P. yoelii infection respond by proliferation in vitro to P. falciparum heat shock proteins (HSPs) of 60 and 70 kDa, suggesting a possible immunological involvement of parasite HSPs in this arm of the cellular immune response during malarial infection in mice. PMID- 9864197 TI - Novel target antigens of the variant-specific immune response to Plasmodium falciparum identified by differential screening of an expression library. AB - A primary infection by the Plasmodium falciparum Palo Alto O and R antigenic variants induces a variant-specific immunity in the Saimiri sciureus monkey. We have shown that these variants express distinct PfEMP1 antigens and differ in their levels of expression of additional antigens, including two conserved erythrocyte membrane-associated proteins, HRP1 and PfEMP3. To identify the antigens eliciting a variant-specific response, we conducted a differential screening of a lambdagt11 library with variant-specific sera. We report here the analysis of the 46 anti-R-specific clones. Two specific targets of the anti-R response were identified: (i) PfEMP3, suggesting that immunogenicity of this antigen is modulated by its relative abundance in different variants, and (ii) Asn-rich motifs. Most anti-R-specific clones, derived from so-far-undescribed genes, were detected by a cross-reaction on poly(Asn) stretches, as indicated by elimination of the signal after absorption on Asn-rich sequences. Reverse transcription-PCR (RT-PCR) showed that expression of the gene defined by clone 13 was R specific. Pepscan analysis of clone 13 identified three Asn-rich polypeptides and one unique peptide reacting specifically with antibodies eluted from the R-infected erythrocyte surface. Antisera raised to the unique peptide reacted with an R-specific protein. Attempts to demonstrate that clone 13 was derived from a var gene by using PCRs combining clone 13 and var-derived primers were unsuccessful. The var genes expressed by O and R parasites were identified not by this strategy but by RT-PCR with var-specific primers. This work has provided novel insights into immunity to antigenic variants and has identified a novel gene switched on during antigenic variation. PMID- 9864198 TI - Mycobacterium tuberculosis catalase and peroxidase activities and resistance to oxidative killing in human monocytes in vitro. AB - Mycobacterium tuberculosis has a relatively high resistance to killing by hydrogen peroxide and organic peroxides. Resistance may be mediated by mycobacterial catalase-peroxidase (KatG) and possibly by alkyl hydroperoxide reductase (AhpC). To determine the interrelationship between sensitivity to H2O2, catalase and peroxidase activities, and bacillary growth rates measured both intracellularly in human monocytes and in culture medium, we examined one laboratory strain, two clinical isolates, and three recombinant strains of M. tuberculosis with differing levels of KatG and AhpC. Five of the mycobacterial strains had intracellular doubling times of 27 to 32 h, while one KatG-deficient clinical isolate (ATCC 35825) doubled in approximately 76 h. Killing of mycobacteria by exogenously added H2O2 was more pronounced for intracellular bacilli than for those bacilli derived from disrupted monocytes. Strains with no detectable KatG expression or catalase activity were relatively sensitive to killing (43 to 67% killing) by exogenous H2O2. However, once even minimal catalase activity was present, mycobacterial catalase activity over a 10-fold range (0.56 to 6.2 U/mg) was associated with survival of 85% of the bacilli. Peroxidase activity levels correlated significantly with resistance of the mycobacterial strains to H2O2-mediated killing. An endogenous oxidative burst induction by 4beta-phorbol 12beta-myristate 13alpha-acetate treatment of infected monocytes reduced the viability of the KatG null strain (H37Rv Inhr) but not the KatG-overexpressing strain [H37Rv(pMH59)]. These results suggest that mycobacterial resistance to oxidative metabolites (including H2O2 and other peroxides) may be an important mechanism of bacillary survival within the host phagocyte. PMID- 9864199 TI - Localization of the intracellular activity domain of Pasteurella multocida toxin to the N terminus. AB - We have shown that Pasteurella multocida toxin (PMT) directly causes transient activation of Gqalpha protein that is coupled to phosphatidylinositol-specific phospholipase Cbeta1 in Xenopus oocytes (B. A. Wilson, X. Zhu, M. Ho, and L. Lu, J. Biol. Chem. 272:1268-1275, 1997). We found that antibodies directed against an N-terminal peptide of PMT inhibited the toxin-induced response in Xenopus oocytes, but antibodies against a C-terminal peptide did not. To test whether the intracellular activity domain of PMT is localized to the N terminus, we conducted a deletion mutational analysis of the PMT protein, using the Xenopus oocyte system as a means of screening for toxin activity. Using PCR and conventional cloning techniques, we cloned from a toxinogenic strain of P. multocida the entire toxA gene, encoding the 1,285-amino-acid PMT protein, and expressed the recombinant toxin as a His-tagged fusion protein in Escherichia coli. We subsequently generated a series of N-terminal and C-terminal deletion mutants and expressed the His-tagged PMT fragments in E. coli. These proteins were screened for cytotoxic activity on cultured Vero cells and for intracellular activity in the Xenopus oocyte system. Only the full-length protein without the His tag exhibited activity on Vero cells. The full-length PMT and N-terminal fragments containing the first 500 residues elicited responses in oocytes, but the C terminal 780 amino acid fragment did not. Our results confirm that the intracellular activity domain of PMT is localized to the N-terminal 500 amino acids of the protein and that the C terminus is required for entry into cells. PMID- 9864200 TI - Campylobacter jejuni-stimulated secretion of interleukin-8 by INT407 cells. AB - Incubation of INT407 cells with various clinical isolates of Campylobacter jejuni resulted in secretion of interleukin-8 (IL-8) at levels ranging from 96 to 554 pg/ml at 24 h. The strains which produced the highest levels of IL-8 secretion were 81-176 and BT44. Induction of IL-8 secretion required live cells of 81-176 and was dependent on de novo protein synthesis. Site-specific mutants of 81-176, which were previously shown to be defective in adherence and invasion, resulted in reduced levels of secretion of IL-8, and cheY mutants of strains 81-176 and 749, which are hyperadherent and hyperinvasive, resulted in higher levels of IL-8 secretion. Another mutant of 81-176, which adheres at about 43% of the wild-type levels but is noninvasive, also showed marked reduction in IL-8 levels, suggesting that invasion is necessary for high levels of IL-8 secretion. When gentamicin was added to INT407 cells at 2 h after infection with 81-176, IL-8 secretion 22 h later was equivalent to that of controls without gentamicin, suggesting that the events which trigger induction and release of IL-8 occur early in the interactions of bacteria and eukaryotic cells. PMID- 9864201 TI - Effect of melanin and carotenoids of Exophiala (Wangiella) dermatitidis on phagocytosis, oxidative burst, and killing by human neutrophils. AB - The black yeast Exophiala (Wangiella) dermatitidis is an increasingly recognized pathogen and a leading cause of severe pheohyphomycosis. Melanin is thought to contribute to the virulence of E. dermatitidis. Whereas the synthesis and the redox properties of melanin have been studied intensively, the influence of melanin and carotenoids on the phagocytosis, the oxidative burst, and the killing of E. dermatitidis by human neutrophils has not been studied. To study their effects on these phenomena, we applied a combination of flow cytometry and a colony-count-dependent method. Using E. dermatitidis wild-type strain 8565 and several melanin-deficient mutants that have been described previously, we demonstrate that melanin prevents this pathogen from being killed in the phagolysosome of the neutrophils. Melanin did not influence the phagocytosis or the oxidative burst of the neutrophils involved. The carotenoids torulene and torularhodine were not found to contribute to the prevention of killing. The ability of E. dermatitidis to block the effects of the neutrophil oxidative burst may critically impair the potential of the host to sufficiently eliminate this fungal pathogen and thus may play an important role in the pathogenesis of phaeohyphomycosis. PMID- 9864202 TI - Comparison of surface proteins of Anaplasma marginale grown in tick cell culture, tick salivary glands, and cattle. AB - Anaplasma marginale, a tick-borne rickettsial pathogen of cattle, infects bovine erythrocytes, resulting in mild to severe hemolytic disease that causes economic losses in domestic livestock worldwide. Recently, the Virginia isolate of A. marginale was propagated in a continuous tick cell line, IDE8, derived from embryonic Ixodes scapularis. Development of A. marginale in cell culture was morphologically similar to that described previously in ticks. In order to evaluate the potential of the cell culture-derived organisms for use in future research or as an antigen for serologic tests and vaccines, the extent of structural conservation of the major surface proteins (MSPs) between the cell culture-derived A. marginale and the bovine erythrocytic stage, currently the source of A. marginale antigen, was determined. Structural conservation on the tick salivary-gland stage was also examined. Monoclonal and monospecific antisera against MSPs 1 through 5, initially characterized against erythrocyte stages, also reacted with A. marginale from cell culture and tick salivary glands. MSP1a among geographic A. marginale isolates is variable in size because of different numbers of a tandemly repeated 28- or 29-amino-acid peptide. The cell culture derived A. marginale maintained the same-size MSP1a as that found on the Virginia isolate of A. marginale in bovine erythrocytes and tick salivary glands. Although differences were observed in the polymorphic MSP2 antigen between culture and salivary-gland stages, MSP2 did not appear to vary, by two-dimensional gel electrophoresis, during continuous passage in culture. These data show that MSPs of erythrocyte-stage A. marginale are present on culture stages and may be structurally conserved during continuous culture. The presence of all current candidate diagnostic and vaccine antigens suggests that in vitro cultures are a valuable source of rickettsiae for basic research and for the development of improved diagnostic reagents and vaccines against anaplasmosis. PMID- 9864203 TI - Catecholamine oxidative products, but not melanin, are produced by Cryptococcus neoformans during neuropathogenesis in mice. AB - Melanin has been proposed as a virulence factor in Cryptococcus neoformans, but its presence has not been shown unambiguously in vivo. Validated methods used previously to show production of cryptococcal eumelanin pigment in vitro (P. R. Williamson, K. Wakamatsu, and S. Ito, J. Bacteriol. 180:1570-1572, 1998) were used to assess for production of laccase-derived products in mouse brain of the Lacc+ strains, 2E-TUC, H99 (serotype A), and ATCC 34873 (serotype D), and the Lacc- strain, 2E-TU. Pyrrole-2,3, 5-tricarboxylic and pyrrole-2,3-dicarboxylic acid, specific degradation products of catecholamine derivatives such as melanin, were found in all Lacc+ strains, but not in the Lacc- strain, 2E-TU. However, the presence of melanin pigment itself could not be demonstrated in the same cells. Lack of the specific degradation products aminohydroxyphenylalanine and aminohydroxyphenylethylamine in Lacc+ strains upon hydriodic acid hydrolysis showed that pheomelanin was also not produced by the fungus in vivo. These are the first data to support the generation of catecholamine oxidation products by C. neoformans in vivo, but they do not support postenzymatic polymerization of these products to form typical eumelanin, as previously proposed. PMID- 9864204 TI - Immunogenicity of intranasally administered meningococcal native outer membrane vesicles in mice. AB - Colonization of the human nasopharyngeal region by Neisseria meningitidis is believed to lead to natural immunity. Although the presence of bactericidal antibody in serum has been correlated with immunity to meningococcal disease, mucosal immunity at the portal of entry may also play an important role. This study was undertaken to examine in mice the possibility of safely using native outer membrane vesicles (NOMV) not exposed to detergent as an intranasal (i.n.) vaccine. The mucosal and systemic responses of mice to intranasal and intraperitoneal (i.p.) vaccination with NOMV were compared over a range of doses from 0.1 to 20 microgram. Intranasal vaccination of mice with NOMV induced a strong systemic bactericidal antibody response, as well as a strong local immunoglobulin A immune response in the lung as determined by assay of lung lavage fluid by enzyme-linked immunosorbent assay and lung antibody secreting cells by enzyme-linked immunospot assay. However, 8- to 10-fold-higher doses of NOMV were required i.n. compared to i.p. to elicit an equivalent bactericidal antibody response in serum. Some NOMV vaccine was aspirated into the lungs of mice during i.n. immunization and resulted in an acute inflammatory response that peaked at 1 to 2 days postimmunization and was cleared by day 7. These results indicate that i.n. delivery of meningococcal NOMV in mice is highly effective in eliciting the production of both a mucosal immune response and a systemic bactericidal antibody response. PMID- 9864205 TI - Expression of the EspB protein of enteropathogenic Escherichia coli within HeLa cells affects stress fibers and cellular morphology. AB - The EspB protein of enteropathogenic Escherichia coli (EPEC) is essential for the signaling events that lead to the accumulation of actin beneath intimately attached bacteria, a process that is known as the attaching and effacing effect. EspB is targeted to the host cell cytoplasm by a type III secretion apparatus. To determine the effect of intracellular EspB on the host cell cytoskeleton, we transfected HeLa cells with a plasmid containing the espB gene under the control of an inducible eukaryotic promoter. A HeLa cell clone that expressed espB mRNA and EspB protein after induction was selected for further study. The expression of EspB in these cells caused a dramatic change in cell morphology and a marked reduction in actin stress fibers. Cells expressing EspB were significantly impaired in their ability to support invasion by EPEC and Salmonella typhimurium. However, the expression of EspB within host cells could not compensate for the lack of EspB expression by an espB mutant strain of EPEC to restore attaching and effacing activity. These studies suggest that EspB is a cytoskeletal toxin that is translocated to the host cell cytoplasm, where it causes a redistribution of actin. PMID- 9864206 TI - Streptococcal pyrogenic exotoxin B induces apoptosis and reduces phagocytic activity in U937 cells. AB - Treatment of U937 human monocyte-like cells with Streptococcus pyogenes led to an induction of apoptosis in these cells. A comparison between the wild-type strain and its isogenic protease-negative mutant indicated that the production of streptococcal pyrogenic exotoxin B (SPE B), a cysteine protease, caused a greater extent of apoptosis in U937 cells. Further study using purified SPE B showed that this protease alone could induce U937 cells to undergo apoptosis, which was characterized by morphologic changes, DNA fragmentation laddering on the gel, and an increase in the percentages of hypodiploid cells. The protease activity of SPE B was required for apoptosis to proceed, since treatment with cysteine protease inhibitor E64 or heat inactivation abrogated this death-inducing effect. The SPE B-induced apoptosis pathway was interleukin-1beta converting enzyme (ICE) family protease dependent. Further experiments showed that the phagocytic activity of U937 cells was reduced by SPE B. Treatment with E64 and heat inactivation both abrogated this phagocytosis-inhibitory effect. Taken together, the present data show that SPE B not only possesses the ability to induce apoptosis in monocytic cells but also helps bacteria to resist phagocytosis by host cells. PMID- 9864207 TI - Expression of listeriolysin O and ActA by intracellular and extracellular Listeria monocytogenes. AB - Listeria monocytogenes requires listeriolysin O (LLO) and ActA, the products of hly and actA, respectively, to establish a productive intracellular infection. LLO is essential for vacuolar lysis and entry into the cytosol, while ActA is required for bacterial spread to adjacent cells. We have used a transcriptional reporter gene system to compare the expression of actA and hly during intracellular growth to that during growth in broth cultures. The hly and actA genes were transcriptionally fused to Escherichia coli lacZ and Bacillus pumilus cat-86 (cat), and the fusions were integrated in single copies into the L. monocytogenes chromosome. A chloramphenicol resistance assay indicated that the hly fusion but not the actA fusion was significantly activated in Luria-Bertani (LB) broth, and this finding correlated with LLO and ActA levels detectable in broth cultures. Quantitation of promoter activity on the basis of beta galactosidase activity revealed up to 10-fold-higher level of expression of the hly fusion relative to the actA fusion in LB broth. In contrast, both fusions were active in the cytosol of J774 cells, and the activity of the actA fusion was approximately 3-fold higher than that of the hly fusion under these conditions. However, quantitative immunoprecipitation of ActA and LLO from infected J774 cells demonstrated approximately 70-fold more cytosolic ActA than cytosolic LLO. Finally, in comparison to induction in broth cultures, actA was highly induced (226-fold) and hly was moderately induced (20-fold) in J774 cells. Collectively, these results indicate that actA and hly are differentially regulated in response to the growth environment and that both genes are preferentially expressed during intracellular growth. Further, while the lower level of production of ActA than of LLO in broth can be accounted for by transcriptional regulation, the relative abundance of intracellular ActA compared to that of intracellular LLO is a function of additional, possibly host-mediated, factors. PMID- 9864209 TI - Resurgent Vibrio cholerae O139: rearrangement of cholera toxin genetic elements and amplification of rrn operon. AB - The unprecedented genesis of a novel non-O1 Vibrio cholerae strain belonging to serogroup O139, which caused an epidemic in late 1992 in the Indian subcontinent, and its subsequent displacement by El Tor O1 vibrios after 18 months initiated a renewed investigation of the aspects of the organism that are related to pathogenesis. The reappearance of V. cholerae O139 with altered antibiotic sensitivity compared to O139 Bengal (O139B) in late 1996 has complicated the epidemiological scenario of V. cholerae and has necessitated an examination of possible rearrangements in the genome underlying such rapid changes in the phenotypic traits. With a view to investigating whether the phenotypic changes that have occurred are associated with alteration in the genome, the genome of the resurgent V. cholerae O139 (O139R) strains were examined. Pulsed-field gel electrophoresis analysis of NotI- and SfiI-digested genomic DNA of O139R isolates showed restriction fragment length polymorphism including in the cholera toxin (CTX) genetic element locus and with O139B isolates. Analyses of the organization of the CTX genetic elements in O139R strains showed that in contrast to two copies of the elements connected by two direct-repeat sequences (RS) in most of the genomes of O139B isolates, the genomes of all O139R strains examined, except strain AS192, have three such elements connected by a single RS. While the RS present in the upstream of the CTX genetic elements in the genome of O139R is of O139B origin, the RS connecting the cores of the elements has several new restriction sites and has lost the BglII site which is supposed to be conserved in all O1 strains and O139B. The endonuclease I-CeuI, which has sites only in the rrn operons in the genomes of all organisms examined so far, has 10 sites in the genomes of O139R strains, compared to 9 in the genomes of O139B strains. The recent isolates of V. cholerae O139 have thus gained one rrn operon. This variation in the number of rrn operons within a serogroup has not been reported for any other organism. The results presented in this report suggest that like the pathogenic El Tor O1 strains, the genomes of O139 strains are undergoing rapid alterations. PMID- 9864208 TI - Induction of pro- and anti-inflammatory cytokines by Borrelia burgdorferi lipoproteins in monocytes is mediated by CD14. AB - We previously showed that heat-killed Borrelia burgdorferi spirochetes and lipidated outer surface protein A (L-OspA) stimulated the in vitro production of interleukin-10 (IL-10) in peripheral blood mononuclear cells (PBMC) from uninfected humans and rhesus monkeys (G. Giambartolomei et al., Infect. Immun. 66:2691-2697, 1998). Here we demonstrate that uninfected human peripheral blood monocytes, but not B or T cells, are the cells that transcribe the IL-10 cytokine gene in response to heat-killed B. burgdorferi. B. burgdorferi similarly induced an upregulation of the IL-1beta and IL-6 cytokine genes in monocytes and the production of IL-10 and IL-6 in culture supernatants of the human monocytic cell line THP-1. Purified L-OspA (but not unlipidated OspA [U-OspA] or U-OspC) also stimulated the production of both cytokines in THP-1 cells in a dose-dependent fashion, suggesting that acylation of the OspA protein molecule is required for the production of both anti- and pro-inflammatory cytokines in naive monocytes. A lipohexapeptide that contained the tripalmitoyl-modified cysteine motif (Pam3Cys Hex) of B. burgdorferi lipoproteins but with an arbitrary peptide sequence had the same effect. Monoclonal antibodies (MAbs) MY4 and 60bca, both of which bind to CD14 and are known to block lipopolysaccharide (LPS)-mediated cytokine production, were able to block L-OspA-mediated IL-10 and IL-6 cytokine production. In contrast, MAb 26ic, which also binds to CD14 but does not block LPS function, failed to inhibit L-OspA-mediated cytokine production. These data suggest that activation of monocytes and production of both anti- and pro inflammatory cytokines induced by lipoproteins proceeds via the CD14 receptor. LPS binding protein was not required for OspA-induced cytokine production. Our results demonstrate that pro- and anti-inflammatory cytokines induced by B. burgdorferi lipoproteins in PBMC are produced by monocytes and that lipoprotein and LPS signaling pathways share at least the initial signaling event that involves the CD14 receptor. PMID- 9864210 TI - Bovine CD4(+) T-lymphocyte clones specific for rhoptry-associated protein 1 of Babesia bigemina stimulate enhanced immunoglobulin G1 (IgG1) and IgG2 synthesis. AB - Optimal protective immunity against babesial infection is postulated to require both complement-fixing and opsonizing antibodies in addition to gamma interferon (IFN-gamma)-mediated macrophage activation. The rhoptry-associated protein 1 (RAP 1) of Babesia bigemina induces partial protective immunity and is a candidate vaccine antigen. Previous studies demonstrated that cattle immunized with native protein that were subsequently protected against challenge had a strong IFN-gamma and weaker interleukin-4 (IL-4) response in immune lymph node lymphocytes that reflected the cytokine profile of the majority of CD4(+) T-cell clones obtained from peripheral blood. RAP-1-specific T helper (Th) cell clones that coexpress IFN-gamma and IL-4 are typical of numerous parasite-specific clones examined. However, the function of such cells as helper cells to enhance immunoglobulin secretion by bovine B cells has not been reported. In cattle, both immunoglobulin G1 (IgG1) and IgG2 can fix complement, but IgG2 is the superior opsonizing subclass. Therefore, studies were undertaken to ascertain the functional relevance of RAP-1-specific, CD4(+) Th0 cells as helper cells to enhance IgG1 and/or IgG2 production by autologous B lymphocytes. For comparison, Th0 clones specific for the metazoan parasite Fasciola hepatica that expressed relatively more IL-4 than the B. bigemina-specific Th cells were similarly assayed. B. bigemina RAP-1-specific clones could enhance production of both IgG1 and IgG2 by autologous B cells, whereas Th cell clones specific for F. hepatica enhanced predominantly IgG1 production. The capacity to enhance IgG2 production was associated with production of IFN-gamma by Th cells cocultured with B cells, antigen, and IL-2. The in vitro helper T-cell activity of these T-cell clones was representative of the in vivo serologic responses, which were composed of a mixed IgG1-IgG2 response in B. bigemina RAP-1 immune cattle and a biased IgG1 response in F. hepatica-immune cattle. PMID- 9864211 TI - Identification of an intestinal neutral glycosphingolipid as a phenotype-specific receptor for the K88ad fimbrial adhesin of Escherichia coli. AB - In this study, we identified a receptor for the K88ad fimbrial adhesin of Escherichia coli in neutral glycosphingolipid preparations from intestinal epithelial cells of K88ad-adhesive pigs, which was absent in preparations from K88ad-nonadhesive pigs. Neither K88ab nor K88ac adhesin variants bound to this neutral glycosphingolipid. Because this receptor is an intestinal glycosphingolipid that binds K88ad adhesin, it has been designated IGLad. Carbohydrate compositional analysis of a partially purified preparation of IGLad identified galactose, glucose, and N-acetylglucosamine in a ratio of 1.5:1.0:0.5 as the major monosaccharides. Preliminary characterization experiments using lectins showed that IGLad contains the terminal glycanic structure Galbeta1 4GlcNAc. Removal of terminal beta-linked galactose residues from IGLad decreased the recognition of IGLad by the K88ad adhesin, indicating that terminal beta linked galactose is an essential component of the K88ad adhesin recognition site on IGLad. Studies with purified glycosphingolipid standards demonstrated that K88ad adhesin binds to neolactotetraosylceramide (nLc4Cer) (Galbeta1-4GlcNAcbeta1 3Galbeta1-4Glcbeta1-1Cer) , lactotriosylceramide (GlcNAcbeta1-3Galbeta1-4Glcbeta1 1Cer) and lactotetraosylceramide (Galbeta1-3GlcNAcbeta1-3Galbeta1-4Glcbeta1-1Cer) . Based on these studies, IGLad appears to be nLc4Cer. PMID- 9864212 TI - The immunoglobulin (IgG) antibody response to OspA and OspB correlates with severe and prolonged Lyme arthritis and the IgG response to P35 correlates with mild and brief arthritis. AB - In an effort to implicate immune responses to specific Borrelia burgdorferi proteins that may have a role in chronic Lyme arthritis, we studied the natural history of the antibody response to B. burgdorferi in serial serum samples from 25 patients monitored throughout the course of Lyme disease. In these patients, the immunoglobulin G (IgM) and IgG antibody responses to 10 recombinant B. burgdorferi proteins, determined during early infection, early arthritis, and maximal arthritis, were correlated with the severity and duration of maximal arthritis. The earliest responses were usually to outer surface protein C (OspC), P35, P37, and P41; reactivity with OspE, OspF, P39, and P93 often developed weeks later; and months to years later, 64% of patients had responses to OspA and OspB. During early infection and early arthritis, the levels of IgG antibody to P35 correlated inversely with the subsequent severity or duration of maximal arthritis. In contrast, during periods of maximal arthritis, the levels of IgG antibody to OspA and OspB, especially to a C-terminal epitope of OspA, correlated directly with the severity and duration of arthritis. Thus, the higher the IgG antibody response to P35 earlier in the infection, the milder and briefer the subsequent arthritis, whereas during maximal arthritis, the higher the IgG response to OspA and OspB, the more severe and prolonged the arthritis. PMID- 9864214 TI - Fimbria-mediated enhanced attachment of nontypeable Haemophilus influenzae to respiratory syncytial virus-infected respiratory epithelial cells. AB - Respiratory syncytial virus (RSV) infection is known to predispose children to otitis media and sinusitis due to bacteria such as nontypeable Haemophilus influenzae (NTHI). In this study, we investigated the role of NTHI surface outer membrane protein P5-homologous fimbriae (P5-fimbriae) in attachment to RSV exposed A549 epithelial cells. Analysis by fluorescence flow cytometry showed that a live P5-fimbriated NTHI strain (NTHIF+) attached to a higher proportion of RSV-exposed A549 cells than to control cells (mean, 68% for RSV versus 29% for control; P = 0.008), while attachment of the P5-fimbriae-deficient isogenic mutant strain (NTHIF-) was significantly lower than in control cells and rose only slightly following RSV exposure (mean, 17% for RSV versus 10% for control, P = 0.229). Attachment of NTHIF+ did not correlate with the amount of RSV antigen expressed by A549 cells. Furthermore, paraformaldehyde-fixed NTHIF+ also demonstrated an enhanced binding to RSV-exposed cells. Observations by transmission electronic microscopy showed that the mean number of bacteria attached per 100 RSV-exposed A549 cells was higher for NTHIF+ than NTHIF- (99 versus 18; P < 0.001). No intracellular bacteria were identified. UV-irradiated conditioned supernatants collected from RSV-infected A549 cultures (UV-cRSV) also enhanced the attachment of NTHIF+ to A549, suggesting the presence of a preformed soluble mediator(s) in UV-cRSV that enhances the expression of receptors for P5 fimbriae on A549 cells. In summary, RSV infection significantly enhances NTHI attachment to respiratory epithelial cells. P5-fimbria is the critical appendage of NTHI that participates in this attachment. In clinical settings, blocking of the P5-fimbria-mediated attachment of NTHIF+ by passive or active immunity may reduce the morbidity due to NTHI during RSV infection. PMID- 9864213 TI - Mutagenesis of active-site histidines of Listeria monocytogenes phosphatidylinositol-specific phospholipase C: effects on enzyme activity and biological function. AB - Listeria monocytogenes, a gram-positive facultative intracellular pathogen, produces two distinct phospholipases C. PC-PLC, encoded by plcB, is a broad-range phospholipase, whereas PI-PLC, encoded by plcA, is specific for phosphatidylinositol. It was previously shown that PI-PLC plays a role in efficient escape of L. monocytogenes from the primary phagosome. To further understand the function of PI-PLC in intracellular growth, site-directed mutagenesis of plcA was performed. Two potential active-site histidine residues were mutated independently to alanine, serine, and phenylalanine. With the exception of the activity of the enzyme containing H38F, which was unstable, the PI-PLC enzyme activities of culture supernatants containing each mutant enzyme were <1% of wild-type activity. In addition, the levels of expression of the mutant PI-PLC proteins were equivalent to wild-type expression. Derivatives of L. monocytogenes containing these specific plcA mutations were found to have phenotypes similar to that of the plcA deletion strain in an assay for escape from the primary vacuole, in intracellular growth in a murine macrophage cell line, and in a plaquing assay for cell-to-cell spread. Thus, catalytic activity of PI-PLC is required for all its intracellular functions. PMID- 9864215 TI - Infection-derived Enterococcus faecalis strains are enriched in esp, a gene encoding a novel surface protein. AB - We report the identification of a new cell wall-associated protein of Enterococcus faecalis. Studies on the distribution of the gene encoding this novel surface protein, Esp, reveal a significant (P < 0.001) enrichment in infection-derived E. faecalis isolates. Interestingly, the esp gene was not identified in any of 34 clinical E. faecium isolates or in 4 other less pathogenic enterococcal species tested. Analysis of the structural gene among various E. faecalis isolates reveals the existence of alternate forms of expression of the Esp protein. The deduced primary structure of the Esp protein from strain MMH594, inferred to be 1,873 amino acids (aa) with a predicted mass of approximately 202 kDa, reveals a core region consisting of repeat units that make up 50% of the protein. Esp bears global organizational similarity to the Rib and C alpha proteins of group B streptococci. Identity among Esp, Rib, and C alpha proteins is strikingly localized to a stretch of 13 aa within repeats of similar length. The high degree of conservation of this 13-residue sequence suggests that it plays an important role in the natural selection for this trait among infection-derived E. faecalis and group B streptococcal isolates. PMID- 9864216 TI - Interaction of NK lysin, a peptide produced by cytolytic lymphocytes, with endotoxin. AB - NK lysin is a 9-kDa polypeptide that was originally isolated from porcine intestinal tissue based on its antibacterial activity. It is produced by cytolytic lymphocytes and is cytolytic against a number of different types of tumor cells. Here we report the binding of NK lysin to lipopolysaccharide (LPS) and its anti-LPS activity. NK lysin binds to matrix-coated LPS from Escherichia coli, Pseudomonas aeruginosa, and different strains of Salmonella enterica. Lipid A and polymyxin B inhibited the binding, demonstrating a preferential interaction of NK lysin with the lipid part of LPS. Chromium-labeled lymphoma cells were lysed by NK lysin, and LPS dose-dependently inhibited the cytolysis at equimolar amounts. In the same manner, NK lysin inhibited certain LPS-stimulated effects on mouse bone marrow cells as well as LPS binding to mouse granulocytes. These results suggest that NK lysin may be a another natural LPS-binding protein from lymphocytes that may participate in the endogenous defense response associated with elevated concentrations of LPS. PMID- 9864217 TI - Gamma interferon augments macrophage activation by lipopolysaccharide by two distinct mechanisms, at the signal transduction level and via an autocrine mechanism involving tumor necrosis factor alpha and interleukin-1. AB - When given in the presence of gamma interferon (IFN-gamma), otherwise nontoxic doses of lipopolysaccharide (LPS or endotoxin) become highly lethal for mice. The mechanisms of this synergistic toxicity are not known. We considered the possibility that an interaction between the LPS-induced NF-kappaB and IFN-gamma induced JAK-STAT pathways at the pretranscriptional level may enhance the LPS induced signals. To test this hypothesis, we incubated murine macrophage RAW 264.7 cells with IFN-gamma for 2 h before addition of different doses of LPS. Consistent with the synergistic induction of inducible nitric oxide synthase mRNA and nitric oxide production by a combination of LPS and IFN-gamma, IFN-gamma strongly augmented LPS-induced NF-kappaB activation and accelerated the binding of NF-kappaB to DNA to as early as 5 min. In agreement with this, IFN-gamma pretreatment promoted rapid degradation of IkappaB-alpha but not that of IkappaB beta. Inhibition of protein synthesis during IFN-gamma treatment suppressed LPS initiated NF-kappaB binding. A rapidly induced protein appeared to be involved in IFN-gamma priming. Preincubation of cells with antibodies to tumor necrosis factor alpha or the interleukin-1 receptor partially reduced the priming effect of IFN-gamma. In a complementary manner, LPS enhanced the activation of signal transducing activator of transcription 1 by IFN-gamma. These data suggest novel mechanisms for the synergy between IFN-gamma and LPS by which they cross-regulate the signal-transducing molecules. Through this mechanism, IFN-gamma may transform a given dose of LPS into a lethal stimulus capable of causing sepsis. It may also serve a beneficial purpose by enabling the host to respond quickly to relatively low doses of LPS and thereby activating antibacterial defenses. PMID- 9864218 TI - Influence of the Salmonella typhimurium pathogenicity island 2 type III secretion system on bacterial growth in the mouse. AB - We have investigated the in vivo growth kinetics of a Salmonella typhimurium strain (P11D10) carrying a mutation in ssaJ, a Salmonella pathogenicity island 2 (SPI2) gene encoding a component of a type III secretion system required for systemic growth in mice. Similar numbers of mutant and wild-type cells were recovered from the spleens and livers of BALB/c mice up to 8 h after inoculation by the intraperitoneal route. Thereafter, the numbers of wild-type cells continued to increase logarithmically in these organs, whereas those of P11D10 remained relatively static for several days before being cleared. Gentamicin protection experiments on spleen cell suspensions recovered from infected mice showed that viable intracellular wild-type bacteria accumulated over time but that intracellular P11D10 cells did not. Infection experiments were also performed with wild-type and P11D10 cells carrying the temperature-sensitive plasmid pHSG422 to distinguish between bacterial growth rates and killing in vivo. At 16 h postinoculation there were 10-fold more wild-type cells than mutant cells in the spleens of infected mice, but the numbers of cells of both strains carrying the nonreplicating plasmid were very similar, showing that there was little difference in the degree of killing sustained by the two strains and that the SPI2 secretion system must be required for bacterial replication, rather than survival, in vivo. The SPI2 mutant phenotype in mice is similar to that of strains carrying mutations in the Salmonella virulence plasmid spv genes. To determine if these two sets of genes interact together, a double mutant strain carrying SPI2 and spv mutations was constructed and compared with strains carrying single mutations in terms of virulence attenuation. These experiments failed to provide any evidence showing that the SPI2 and spv gene products interact together as part of the same virulence mechanism. PMID- 9864220 TI - Type-specific contributions to chromosome size differences in Escherichia coli. AB - The Escherichia coli genome varies in size from 4.5 to 5.5 Mb. It is unclear whether this variation may be distributed finely throughout the genome or is concentrated at just a few chromosomal loci or on plasmids. Further, the functional correlates of size variation in different genome copies are largely unexplored. We carried out comparative macrorestriction mapping using rare restriction-site alleles (made with the Tn10dRCP2 family of elements, containing the NotI, BlnI, I-CeuI, and ultra-rare-cutting I-SceI sites) among the chromosomes of laboratory E. coli K-12, newborn-sepsis-associated E. coli RS218, and uropathogenic E. coli J96. These comparisons showed just a few large accessory chromosomal segments accounting for nearly all strain-to-strain size differences. Of 10 sepsis-associated and urovirulence genes, previously isolated from the two pathogens by scoring for function, all were colocalized exclusively with one or more of the accessory chromosomal segments. The accessory chromosomal segments detected in the pathogenic strains from physical, macrorestriction comparisons may be a source of new virulence genes, not yet isolated by function. PMID- 9864219 TI - Mechanisms for induction of L-selectin loss from T lymphocytes by a cryptococcal polysaccharide, glucuronoxylomannan. AB - Disseminated cryptococcosis is accompanied by cryptococcal polysaccharides in the serum and the lack of cellular infiltrates in infected tissues. Cryptococcal polysaccharides given intravenously to mice inhibit the influx of T lymphocytes into the sites of cell-mediated immune response. The focus here was to determine whether cryptococcal polysaccharides modulate the expression of molecules, such as L-selectin, that are important in extravasation of T cells. Cryptococcal glucuronoxylomannan (GXM), but not galactoxylomannan or mannoprotein, was found to cause loss of L-selectin from freshly isolated human T cells of both CD4 and CD8 subsets and from Jurkat cells. With the signaling-pathway inhibitors staurosporine (which inhibits protein kinase C) and herbimycin A (which inhibits protein tyrosine kinases), we showed that GXM or the cryptococcal culture filtrate antigen CneF directly induces L-selectin loss from CD4(+) and CD8(+) T cells via a herbimycin A-sensitive pathway(s) presumably involving one or more protein tyrosine kinases but not via a pathway involving protein kinase C. Loss of L-selectin from the T cells before the T cells have a chance to bind to L selectin ligands on endothelial cells would be expected to prevent T-cell migration into inflamed tissues and/or lymph organs. PMID- 9864221 TI - Compensation for decreased expression of B7 molecules on Leishmania infantum infected canine macrophages results in restoration of parasite-specific T-cell proliferation and gamma interferon production. AB - Infection of humans and dogs by Leishmania infantum may result in visceral leishmaniasis, which is characterized by impaired T-cell-mediated immune responses to parasite antigens. Dogs are natural hosts of Leishmania parasites and play an important role in the transmission of the parasites to humans. In an effort to characterize the immune response in dogs infected with this intracellular pathogen, we examined how infection with L. infantum affects canine macrophages and the consequences for T-cell activation in vitro. We showed that the proliferation of T-cell lines to cognate antigen decreases to background levels when infected autologous monocyte-derived macrophages are used as antigen presenting cells (APC). The observed reduction of antigen-specific T-cell proliferation was shown to be dependent on the parasite load and to require cell to-cell interaction of T cells with the infected APC. In addition, we observed a decreased expression of costimulatory B7 molecules on infected monocyte-derived macrophages. The expression of other surface molecules involved in T-cell activation, such as major histocompatibility complex class I and class II, on these cells did not change upon infection, whereas the expression of intracellular adhesion molecule 1 was marginally increased. Compensation for the decreased expression of B7 molecules by the addition of B7-transfected cells resulted in the restoration of cell proliferation and gamma interferon (IFN gamma) production by a Leishmania-specific T-cell line. These results showed that for the activation of parasite-specific canine T cells producing IFN-gamma, which are most likely involved in protective immunity, sufficient expression of B7 molecules on infected macrophages is required. Provision of costimulatory molecules may be an approach for immunotherapy of leishmaniaisis as well as for vaccine development. PMID- 9864222 TI - Modulation of endotoxin- and enterotoxin-induced cytokine release by in vivo treatment with beta-(1,6)-branched beta-(1,3)-glucan. AB - Leukocytes activated by endotoxin or enterotoxins release proinflammatory cytokines, thereby contributing to the cascade of events leading to septic shock. In the present studies, we analyzed the effects of in vivo administration of a soluble immunomodulator, beta-(1,6)-branched beta-(1,3)-glucan (soluble beta glucan), on toxin-stimulated cytokine production in monocytes and lymphocytes isolated from treated mice. In vitro stimulation of lymphocytes isolated from soluble beta-glucan-treated mice with lipopolysaccharide (LPS) resulted in enhanced production of interleukin-6 (IL-6) and suppressed production of tumor necrosis factor alpha (TNF-alpha), while stimulation of these cells with staphylococcal enterotoxin B (SEB) or toxic shock syndrome toxin 1 (TSST-1) resulted in enhanced production of gamma interferon (IFN-gamma) and suppressed production of IL-2 and TNF-alpha compared to that in cells isolated from untreated mice. In vitro stimulation of monocytes isolated from soluble beta glucan-treated mice with LPS also resulted in suppressed TNF-alpha production, while stimulation of these cells with SEB or TSST-1 resulted in suppressed IL-6 and TNF-alpha production compared to that in cells isolated from untreated mice. Thus, the overall cytokine pattern of leukocytes from soluble beta-glucan-treated mice reflects suppressed production of proinflammatory cytokines, especially TNF alpha. Taken together, our results suggest that treatment with soluble beta glucan can modulate the induction cytokines during sepsis, resulting in an overall decrease in host mortality. PMID- 9864223 TI - Existing antilisterial immunity does not inhibit the development of a Listeria monocytogenes-specific primary cytotoxic T-lymphocyte response. AB - Infection of BALB/c mice with Listeria monocytogenes stimulates an antilisterial immune response evident by the appearance of H2-Kd-restricted CD8(+) cytotoxic T lymphocytes (CTLs) specific for the nanomer peptides amino acids (aa) 91 to 99 of listeriolysin O (LLO 91-99) and aa 217 to 225 of the p60 molecule (p60 217-225). We have introduced point mutations at anchor residues within LLO 91-99 (92F) or p60 217-225 (218F), and BALB/c mice infected with L. monocytogenes strains containing these point mutations do not develop CTLs specific for LLO 91-99 or p60 217-225, respectively. We have used these strains to test whether primary CTL responses against L. monocytogenes-derived determinants can be stimulated within an environment of existing antilisterial immunity. We found that the development of a primary L. monocytogenes-specific CTL response is not altered by existing immunity to L. monocytogenes. For example, primary immunization with the p60 218F strain of L. monocytogenes followed by a secondary immunization with wild-type L. monocytogenes results in stimulation of p60 217-225-specific CTLs at primary response levels and LLO 91-99-specific effectors at levels consistent with a memory CTL response. Similarly, primary immunization with the 92F strain of L. monocytogenes followed by a secondary immunization with wild-type L. monocytogenes results in stimulation of LLO 91-99-specific CTLs at primary response levels and p60 217-225-specific effectors at levels consistent with a memory CTL response. These results provide additional support for the use of L. monocytogenes as a recombinant vaccine vector and show that antivector immunity does not inhibit the development of a primary CTL response when the epitope is delivered by L. monocytogenes as the vaccine strain. PMID- 9864224 TI - Effects of site-directed mutagenesis of Escherichia coli heat-labile enterotoxin on ADP-ribosyltransferase activity and interaction with ADP-ribosylation factors. AB - Escherichia coli heat-labile enterotoxin (LT), an oligomeric protein with one A subunit (LTA) and five B subunits, exerts its effects via the ADP-ribosylation of Gsalpha, a guanine nucleotide-binding (G) protein that activates adenylyl cyclase. LTA also ADP-ribosylates simple guanidino compounds (e.g., arginine) and catalyzes its own auto-ADP-ribosylation. All LTA-catalyzed reactions are enhanced by ADP-ribosylation factors (ARFs), 20-kDa guanine nucleotide-binding proteins. Replacement of arginine-7 (R7K), valine-53 (V53D), serine-63 (S63K), valine 97 (V97K), or tyrosine-104 (Y104K) in LTA resulted in fully assembled but nontoxic proteins. S63K, V53D, and R7K are catalytic-site mutations, whereas V97K and Y104K are amino acid replacements adjacent to and outside of the catalytic site, respectively. The effects of mutagenesis were quantified by measuring ADP ribosyltransferase activity (i.e., auto-ADP-ribosylation and ADP-ribosylagmatine synthesis) and interaction with ARF (i.e., inhibition of ARF-stimulated cholera toxin ADP-ribosyltransferase activity and effects of ARF on mutant auto-ADP ribosylation). All mutants were inactive in the ADP-ribosyltransferase assay; however, auto-ADP-ribosylation in the presence of recombinant human ARF6 was detected, albeit much less than that of native LT (Y104K > V53D > V97K > R7K, S63K). Based on the lack of inhibition by free ADP-ribose, the observed auto-ADP ribosylation activity was enzymatic and not due to the nonenzymatic addition of free ADP-ribose. V53D, S63K, and R7K were more effective than Y104K or V97K in blocking ARF stimulation of cholera toxin ADP-ribosyltransferase. Based on these data, it appears that ARF-binding and catalytic sites are not identical and that a region outside the NAD cleft may participate in the LTA-ARF interaction. PMID- 9864225 TI - Polarized helper-T-cell responses against Leishmania major in the absence of B cells. AB - B-cell-to-T-cell signaling can shape helper T (Th) cell responses. During infection with Leishmania major, Th response is critical in determining the outcome of disease. Resistance depends on the generation of a protective Th1 response, while susceptibility is mediated by the generation of a Th2 response. In this study, we determined whether B cells are required for the development of polarized Th1 and Th2 responses during infection with L. major. Mice lacking B cells due to disruption of the immunoglobulin M locus (microMT) were infected with L. major, and disease progression and Th cell development were assessed. On the genetically resistant C57BL background, both wild-type and microMT mice controlled the infection and mounted a Th1 response. On the genetically susceptible BALB/c background, both wild-type and microMT mice were susceptible to infection and generated Th2 responses. Thus, during L. major infection, neither direct antigen presentation or costimulation by B cells nor antibody mediated effector functions are essential for the development of polarized Th responses. PMID- 9864227 TI - Role of gamma interferon in Helicobacter pylori-induced gastric inflammatory responses in a mouse model. AB - The immune responses to Helicobacter pylori infection play important roles in gastroduodenal diseases. The contribution of gamma interferon (IFN-gamma) to the immune responses, especially to the induction of gastric inflammation and to protection from H. pylori infection, was investigated with IFN-gamma gene knockout (IFN-gamma-/-) mice. We first examined the colonizing abilities of eight H. pylori strains with a short-term infection test in order to select H. pylori strains which could colonize the mouse stomach. Only three strains (ATCC 43504, CPY2052, and HPK127) colonized C57BL/6 wild-type mice, although all of the strains except for ATCC 51110 could colonize IFN-gamma-/- mice. The number of H. pylori organisms colonizing the stomach in wild-type mice was lower than that in IFN-gamma-/- mice. Oral immunization with the CPY2052 sonicate and cholera toxin protected against infection with strain CPY2052 in both types of mouse. These findings suggested that IFN-gamma may play a protective role in H. pylori infection, although the degree of its protective ability was estimated to be low. In contrast, in a long-term infection test done to examine the contribution of IFN-gamma to gastric inflammation, CPY2052-infected wild-type mice developed a severe infiltration of mononuclear cells in the lamina propria and erosions in the gastric epithelium 15 months after infection, whereas CPY2052-infected IFN gamma-/- mice showed no inflammatory symptoms. This result clearly demonstrated that IFN-gamma plays an important role in the induction of gastric inflammation caused by H. pylori infection. PMID- 9864226 TI - An extracellular protease of Streptococcus gordonii hydrolyzes type IV collagen and collagen analogues. AB - Streptococcus gordonii is a frequent cause of infective bacterial endocarditis, but its mechanisms of virulence are not well defined. In this study, streptococcal proteases were recovered from spent chemically defined medium (CDM) and fractionated by ammonium sulfate precipitation and by ion-exchange and gel filtration column chromatography. Three proteases were distinguished by their different solubilities in ammonium sulfate and their specificities for synthetic peptides. One of the enzymes cleaved collagen analogs Gly-Pro 4-methoxy-beta naphthylamide, 2-furanacryloyl-Leu-Gly-Pro-Ala (FALGPA), and p phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-Arg (pZ-peptide) and was released from the streptococci while complexed to peptidoglycan fragments. Treatment of this protease with mutanolysin reduced its 180- to 200-kDa mass to 98 kDa without loss of enzymatic activity. The purified protease cleaved bovine gelatin, human placental type IV collagen, and the Aalpha chain of fibrinogen but not albumin, fibronectin, laminin, or myosin. Enzyme activity was inhibited by phenylmethylsulfonyl fluoride, indicating that it is a serine-type protease. Maximum production of the 98-kDa protease occurred during growth of S. gordonii CH1 in CDM containing 0.075% total amino acids at pH 7.0 with minimal aeration. Higher initial concentrations of amino acids prevented the release of the protease without reducing cell-associated enzyme levels, and the addition of an amino acid mixture to an actively secreting culture stopped further enzyme release. The purified protease was stored frozen at -20 degreesC for several months or heated at 50 degreesC for 10 min without loss of activity. These data indicate that S. gordonii produces an extracellular gelatinase/type IV collagenase during growth in medium containing minimal concentrations of free amino acids. Thus, the extracellular enzyme is a potential virulence factor in the amino acid-stringent, thrombotic, valvular lesions of bacterial endocarditis. PMID- 9864228 TI - Cloning and characterization of a novel membrane-associated antigenic protein of Helicobacter pylori. AB - Infection by Helicobacter pylori, a noninvasive bacterium, induces chronic leukocyte infiltration in the stomach by still largely unknown molecular mechanisms. We investigated the possibility that a membrane protein of H. pylori induces an inflammatory reaction in the subepithelial tissue of the stomach. By generating an expression library of H. pylori chromosomal DNA and screening with rabbit antiserum raised to a membrane fraction of H. pylori and sera of infected patients, we cloned a 16.0-kDa protein (HP-MP1) which appeared to attach to the inner membrane of the H. pylori in a homodimeric form. Anti-HP-MP1 antibodies were detected in the sera of infected patients but not in those of uninfected controls. Coincubation of monocytes with recombinant HP-MP1 led to cell activation and production of interleukin-1alpha (IL-1alpha), tumor necrosis factor alpha, IL-8, and macrophage inflammatory protein 1alpha. The results indicate that HP-MP1 is an antigenic membrane-associated protein of H. pylori which potentially activates monocytes. This suggests that HP-MP1 may play roles in the pathogenesis of perpetual tissue inflammation associated with H. pylori infection. PMID- 9864229 TI - Vibrio anguillarum resistance to rainbow trout (Oncorhynchus mykiss) serum: role of O-antigen structure of lipopolysaccharide. AB - The sensitivity of Vibrio anguillarum to the bactericidal effect of rainbow trout serum was investigated with different strains of serogroups O1 and O2a, which are the most frequently found serogroups in clinical outbreaks of vibriosis. All of the V. anguillarum strains were able to activate complement in rainbow trout serum, but smooth strains of V. anguillarum serogroup O1 were resistant to complement-mediated killing in the absence of specific antibodies. In the case of V. anguillarum serogroup O2a strains, 80% of the analyzed strains were resistant to rainbow trout serum even when specific antibodies were present. Analysis of the lipopolysaccharide structures of the tested V. anguillarum strains showed a positive correlation between the O-antigen size of the lipopolysaccharide and resistance to serum killing. The classical complement pathway was responsible for the antibody-dependent serum killing of susceptible V. anguillarum strains. When serum-resistant V. anguillarum serogroup O2a strains were grown in glucose enriched Lennox L broth, they produced lipopolysaccharide molecules with fewer high-molecular-weight O-antigen units than did strains grown in broth without the addition of glucose. Strains grown in glucose-enriched medium became sensitive to rainbow trout serum killing, indicating that the high-molecular-weight O-antigen side chains prevented the activated complement from damaging the bacterium. PMID- 9864230 TI - Saccharomyces boulardii protease inhibits the effects of Clostridium difficile toxins A and B in human colonic mucosa. AB - Saccharomyces boulardii is a nonpathogenic yeast used in the treatment of Clostridium difficile diarrhea and colitis. We have reported that S. boulardii inhibits C. difficile toxin A enteritis in rats by releasing a 54-kDa protease which digests the toxin A molecule and its brush border membrane (BBM) receptor (I. Castagliuolo, J. T. LaMont, S. T. Nikulasson, and C. Pothoulakis, Infect. Immun. 64:5225-5232, 1996). The aim of this study was to further evaluate the role of S. boulardii protease in preventing C. difficile toxin A enteritis in rat ileum and determine whether it protects human colonic mucosa from C. difficile toxins. A polyclonal rabbit antiserum raised against purified S. boulardii serine protease inhibited by 73% the proteolytic activity present in S. boulardii conditioned medium in vitro. The anti-protease immunoglobulin G (IgG) prevented the action of S. boulardii on toxin A-induced intestinal secretion and mucosal permeability to [3H]mannitol in rat ileal loops, while control rabbit IgG had no effect. The anti-protease IgG also prevented the effects of S. boulardii protease on digestion of toxins A and B and on binding of [3H]toxin A and [3H]toxin B to purified human colonic BBM. Purified S. boulardii protease reversed toxin A- and toxin B-induced inhibition of protein synthesis in human colonic (HT-29) cells. Furthermore, toxin A- and B-induced drops in transepithelial resistance in human colonic mucosa mounted in Ussing chambers were reversed by 60 and 68%, respectively, by preexposing the toxins to S. boulardii protease. We conclude that the protective effects of S. boulardii on C. difficile-induced inflammatory diarrhea in humans are due, at least in part, to proteolytic digestion of toxin A and B molecules by a secreted protease. PMID- 9864231 TI - Influence of acute-phase parasite load on pathology, parasitism, and activation of the immune system at the late chronic phase of Chagas' disease. AB - To obtain low and high parasite loads in the acute phase of Chagas' disease, A/J mice were infected with 10(3) or 10(5) Trypanosoma cruzi trypomastigotes of the Y strain and treated on day 6 with benznidazol. One year later, chronically infected mice were screened for subpatent parasitemias, tissue pathology, and immune response. Mice infected with the high parasite inoculum showed higher levels of chronic parasitemias, heart and striated muscle inflammation, and activation of the immune system than did mice infected with the low inoculum. Concerning the activation of the immune system, the main findings for high-dose infected mice were (i) increased numbers of splenocytes, with preferential expansion of CD8(+) and B220(-) CD5(-) cells, many of them bearing a macrophage phenotype; (ii) higher frequencies of B (B220(+)), CD4(+), and CD8(+) large lymphocytes; (iii) a shift of CD4(+) cells towards a CD45RBLow phenotype; (iv) increased frequencies of both CD45RBLow and CD45RBHigh large CD4(+) cells; (v) augmented numbers of total immunoglobulin (Ig)-secreting cells, with predominance of IgG2a-producing cells; and (vi) increased production of gamma interferon and interleukin 4. In addition, these mice presented lower IgM and higher IgG2a and IgG1 parasite-specific serum antibody levels. Our results indicate that the parasite load at the acute phase of T. cruzi infection influences the activation of the immune system and development of Chagas' disease pathology at the late chronic phase of the disease. PMID- 9864232 TI - The polysaccharide portion of lipopolysaccharide regulates antigen-specific T cell activation via effects on macrophage-mediated antigen processing. AB - The lipopolysaccharide (LPS) structure of Salmonella typhimurium has been correlated with the virulence of wild-type strain LT2. Mutants of LT2 with truncated polysaccharide portions of LPS are less virulent than strains with a complete LPS structure. Polyclonal T cells and monoclonal T-cell hybridomas were more reactive to heat-killed rough mutants than to heat-killed smooth strains, as measured by interleukin-2 (IL-2) production. Using a large panel of strains with truncated LPS molecules, we found that T-cell reactivity decreased with certain lengths of polysaccharide. The decreased response was not due to differential phagocytic uptake, IL-12 production, or major histocompatibility complex class II surface expression by macrophages. Also, LT2 did not mediate any global suppression since addition of LT2 did not diminish the response of T cells specific for antigens unrelated to Salmonella. In an experiment in which processing times were varied, we found that antigens from rough strains were processed and presented more quickly than those associated with smooth strains. At longer processing times, epitopes from LT2 were presented well. We hypothesize that the slower antigen processing and presentation of wild-type Salmonella may be caused by masking of surface antigens by the longer polysaccharide portion of smooth LPS. This blocking of effective antigen presentation may contribute to the virulence of Salmonella. PMID- 9864234 TI - Helicobacter pylori infection in immunized mice lacking major histocompatibility complex class I and class II functions. AB - The role of major histocompatibility complex (MHC) class I- and class II restricted functions in Helicobacter pylori infection and immunity upon oral immunization was examined in vivo. Experimental challenge with H. pylori SS1 resulted in significantly greater (P 0.05). Three of the five animals in this group also had low parasitemia (peak parasitemia, 100-fold when tissues from 3-week-infected mice were compared with those from 10-month-infected mice. However, even at the lower level, parasites were clearly recognizable in sections of skeletal muscle and bladder at the 10 month time point. Inflammation remained robust in skeletal muscle, bladder, and sciatic nerve despite the near disappearance of parasites, suggesting three possibilities: exuberant host reactions to the few remaining parasites, autoimmune inflammation, or reactions to retained parasite antigens in the tissues. PMID- 9864244 TI - Five homologous repeats of the protein G-related protein MIG cooperate in binding to goat immunoglobulin G. AB - Protein MIG, from Streptococcus dysgalactiae, binds alpha2-macroglobulin and immunoglobulin G (IgG). MIG-derived fusion proteins with one to five IgG-binding repeats differed up to 72,000- fold in avidity for goat IgG, indicating a considerable cooperativity of the repeats. Significant sequence variation in the IgG-binding repeats was recognized. Protein MIG interacted with goat IgG1 via both the Fc and Fab parts. PMID- 9864245 TI - Increased levels of soluble CD14 in sera of periodontitis patients. AB - Soluble CD14 (sCD14) mediates the response to lipopolysaccharide (LPS) in cells lacking membrane-bound CD14. We determined sCD14 concentrations in the sera of 38 periodontitis patients and 25 healthy controls by enzyme-linked immunosorbent assay. The sCD14 levels in the sera of patients with periodontitis were significantly higher than those of healthy subjects and decreased after treatment. Enhanced levels of sCD14 in serum may contribute to the host response to LPS in periodontitis. Furthermore, we showed in vitro that addition of LPS enhanced the release of sCD14 by monoblastic U937 cells treated with 1alpha,25 dihydroxyvitamin D3. Thus, increased sCD14 levels in periodontitis patients may be due to chronic exposure to LPS. PMID- 9864246 TI - Antisense inhibition of expression of cysteine proteinases affects Entamoeba histolytica-induced formation of liver abscess in hamsters. AB - Trophozoites of virulent Entamoeba histolytica transfected with the antisense gene encoding cysteine proteinase 5 (CP5) have only 10% of the CP activity but retain their cytopathic activity on mammalian monolayers. In the present study we found that the transfected trophozoites with low levels of CP activity were incapable of inducing the formation of liver lesions in hamsters. PMID- 9864247 TI - Soluble tumor necrosis factor alpha receptors in sera from leprosy patients. AB - Serum levels of soluble tumor necrosis factor alpha receptor I (sTNF-RI) were elevated in patients with lepromatous (LL) reactional-state type II leprosy, and sTNF-RII levels were increased in patients with full tuberculoid (TT) or LL type II leprosy. The sTNF-R in sera from patients with type II leprosy, but not other forms of leprosy, inhibited recombinant TNF cytolytic activities in vitro. This suggests that sTNF-R regulatory activities are partially impaired in patients with leprosy. PMID- 9864248 TI - Tonsillar application of formalin-killed cells of Streptococcus sobrinus reduces experimental dental caries in rabbits. AB - Living Streptococcus sobrinus cells were orally inoculated into nonimmune rabbits and rabbits immunized with formalin-killed cells of S. sobrinus through tonsillar application to examine the anticaries potential of this method of immunization. The living S. sobrinus cell numbers and the caries areas in the rabbits immunized by tonsillar application decreased to a level one-fifth of that in nonimmune rabbits. PMID- 9864249 TI - Virulence properties of Escherichia coli 83972, a prototype strain associated with asymptomatic bacteriuria. AB - Little is known about bacteria associated with asymptomatic bacteriuria (ABU) with regard to urinary tract colonization mechanisms. In this study, virulence properties of Escherichia coli 83972, a strain that was isolated from a clinical ABU episode, were examined. The genetic potential for expression of P and type 1 pili was demonstrated, and DNA sequences related to type 1C and G (UCA) pilus genes were also detected. However, E. coli 83972 did not express D-mannose resistant or D-mannose-sensitive hemagglutination after growth under standard conditions in vitro or upon isolation from the urine of colonized test subjects. Limited uroepithelial cell adherence was observed in vivo, and weak D-mannose sensitive hemagglutination was detected after extended growth in urine in vitro. PMID- 9864250 TI - Nonrestricted differential intoxication of cells by pertussis toxin. AB - After uptake and retrograde transport pertussis toxin acts by ADP-ribosylating alpha-Gi proteins. We show that uptake via many different receptor proteins followed by retrograde transport and intoxication is not restricted to a particular cell type. The efficiency of cellular intoxication, however, was found to be cell type dependent. PMID- 9864251 TI - Glucose 6-phosphate dehydrogenase is required for Salmonella typhimurium virulence and resistance to reactive oxygen and nitrogen intermediates. AB - Salmonella typhimurium zwf mutants lacking glucose 6-phosphate dehydrogenase (G6PD) activity have increased susceptibility to reactive oxygen and nitrogen intermediates as well as attenuated virulence in mice. Abrogation of the phagocyte respiratory burst oxidase during experimental infection with zwf mutant Salmonella causes a prompt restoration of virulence, while inhibition of inducible nitric oxide synthase results in delayed lethality. These observations suggest that G6PD-dependent bacterial antioxidant defenses play an important pathogenic role during early salmonellosis and additionally may help to antagonize NO-dependent antimicrobial mechanisms later in the course of infection. PMID- 9864252 TI - Cathelicidin gene expression in porcine tissues: roles in ontogeny and tissue specificity. AB - Cathelicidins constitute a family of mammalian antimicrobial peptides that are synthesized in the bone marrow as prepropeptides, stored in neutrophil granules as propeptides, and released as active, mature peptides upon neutrophil degranulation. We investigated the developmental expression of two porcine cathelicidins, PR-39 and protegrin. Both cathelicidins were expressed constitutively in the bone marrow of all pigs at all of the ages tested. Peripheral blood neutrophils from young pigs expressed PR-39 and protegrin mRNA, which were not detectable at 42 days of age. At earlier ages, expression of PR-39 mRNA was detected in the kidney and liver and several lymphoid organs, including the thymus, spleen, and mesenteric lymph nodes, but disappeared at 4 weeks of age. These data provide the first evidence of cathelicidin gene expression in peripheral leukocytes and may indicate a role for these antimicrobial peptides in the development of host defense mechanisms. PMID- 9864253 TI - Killing of Borrelia burgdorferi by antibody elicited by OspA vaccine is inefficient in the absence of complement. AB - A Lyme disease vaccine, based on the Borrelia burgdorferi lipoprotein OspA, has recently undergone phase III trials in humans. The results of one of these trials indicate that vaccine efficacy positively correlates with anti-OspA antibody titer. Spirochete killing within the tick vector midgut, upon which vaccine efficacy appears to depend, may occur chiefly via a mechanism that involves antibody alone, as it has been reported that complement is degraded by tick saliva decomplementing factors. We compared the in vitro killing efficiencies of anti-OspA antibody elicited in rhesus monkeys by the OspA vaccine, in the presence and in the absence of monkey complement. Killing in the absence of complement was between 14 and 3,800 times less efficient than with complement present, depending on the spirochete strain. The relative inefficiency of the complement-independent killing mechanism by anti-OspA antibody may explain why OspA vaccine efficacy is critically dependent on antibody titer. PMID- 9864254 TI - Gamma delta T-cell function in pathogenesis of cerebral malaria in mice infected with Plasmodium berghei ANKA. AB - Mice depleted of gammadelta T cells by monoclonal antibody treatment and infected with Plasmodium berghei ANKA did not develop cerebral malaria (CM). In striking contrast, delta0/0 mice infected with P. berghei developed CM despite their gammadelta T-cell deficiency. gammadelta T cells appear to be essential for the pathogenesis of CM in mice having experienced normal ontogeny but not in mice genetically deprived of gammadelta T cells from the beginning of life. PMID- 9864255 TI - Adhesin expression in matched nasopharyngeal and middle ear isolates of nontypeable Haemophilus influenzae from children with acute otitis media. AB - The HMW1 and HMW2 proteins, Hia, and hemagglutinating pili are important adherence factors in nontypeable Haemophilus influenzae. To gain insight into the relative importance of these adhesins in nasopharyngeal colonization and localized respiratory tract disease, we assessed their expression in matched nasopharyngeal and middle ear isolates of nontypeable H. influenzae from 17 children with acute otitis media. In all patients, including 11 with bilateral disease, the matched isolates were isogenic based on total protein profiles and genomic fingerprints. Of the nasopharyngeal isolates, 14 expressed only HMW1/HMW2 like proteins, 1 expressed only Hia, 1 expressed only pili, and 1 expressed both Hia and pili. Further analysis revealed concordance between nasopharyngeal isolates and the matched middle ear isolates for expression of the HMW1/HMW2-like proteins and Hia. In contrast, in the two children whose nasopharynges were colonized by piliated organisms, the corresponding middle ear isolates were nonpiliated and could not be enriched for piliation. Nevertheless, Southern analysis revealed that these two middle ear isolates contained all five hif genes required for pilus biogenesis and had no evidence of major genetic rearrangement. In summary, the vast majority of isolates of nontypeable H. influenzae associated with acute otitis media express HMW1/HMW2-like proteins, with expression present in both the nasopharynx and the middle ear. A smaller fraction of nasopharyngeal isolates express pili, while isogenic strains recovered from the middle ear are often refractory to enrichment for piliation. We speculate that the HMW adhesins and Hia are important at multiple steps in the pathogenesis of otitis media while pili contribute to early colonization and then become dispensable. PMID- 9864256 TI - Identification and functional characterization of the Neisseria gonorrhoeae lbpB gene product. AB - We cloned lbpB, encoding a predicted 80-kDa lipoprotein, upstream of lbpA. A nonpolar mutant (LbpB- LbpA+) had normal lactoferrin (LF) binding and grew normally with LF as an iron source, whereas LbpB- LbpA- and LbpB+ LbpA- strains had reduced binding of LF and did not grow with LF as an iron source. LbpB bound LF directly in an affinity purification, suggesting that LbpB might play a still uncharacterized role in the LF iron utilization. PMID- 9864257 TI - Response to reactive nitrogen intermediates in Mycobacterium tuberculosis: induction of the 16-kilodalton alpha-crystallin homolog by exposure to nitric oxide donors. AB - In contrast to the apparent paucity of Mycobacterium tuberculosis response to reactive oxygen intermediates, this organism has evolved a specific response to nitric oxide challenge. Exposure of M. tuberculosis to NO donors induces the synthesis of a set of polypeptides that have been collectively termed Nox. In this work, the most prominent Nox polypeptide, Nox16, was identified by immunoblotting and by N-terminal sequencing as the alpha-crystallin-related, 16 kDa small heat shock protein, sHsp16. A panel of chemically diverse donors of nitric oxide, with the exception of nitroprusside, induced sHsp16 (Nox16). Nitroprusside, a coordination complex of Fe2+ with a nitrosonium (NO+) ion, induced a 19-kDa polypeptide (Nox19) homologous to the nonheme bacterial ferritins. We conclude that the NO response in M. tuberculosis is dominated by increased synthesis of the alpha-crystallin homolog sHsp16, previously implicated in stationary-phase processes and found in this study to be a major M. tuberculosis protein induced upon exposure to reactive nitrogen intermediates. PMID- 9864260 TI - Inhibitory action of mibefradil on calcium signaling and aldosterone synthesis in bovine adrenal glomerulosa cells. AB - Mibefradil is a new cardiovascular drug with peculiar Ca++ antagonistic properties. The most remarkable feature of mibefradil is its unique relative selectivity for T type calcium channels, a property that has been proposed to explain in part the beneficial pharmacological and clinical profiles of this drug. In adrenal glomerulosa cells, aldosterone biosynthesis and secretion in response to angiotensin II or extracellular potassium is dependent on a sustained influx of Ca++ through T type Ca++ channels. The effect of mibefradil on the steroidogenic function of glomerulosa cells was therefore investigated. Using the patch clamp technique, we found that mibefradil inhibits selectively and in a concentration-dependent manner (IC50 = 3 microM)++ T type currents in bovine glomerulosa cells. In addition to this tonic (voltage independent) inhibition, the drug also induced a shift of the steady-state inactivation curve of these channels toward hyperpolarized voltages, contributing to its efficacy to prevent Ca++ influx into the cell through T type channels. Concomitantly, mibefradil reduced the cytosolic calcium responses to potassium and angiotensin II (as assessed with fluorescent probes), without affecting the capacitative Ca++ influx, and inhibited pregnenolone and aldosterone formation. This inhibition of steroidogenesis was not exclusively due to mibefradil action on voltage-operated Ca++ channels, because this agent also partially reduced steroid synthesis induced by adrenocorticotropic hormone or forskolin, two activators of the cyclic AMP pathway. In conclusion, mibefradil is highly effective in adrenal glomerulosa cells in reducing T type channel activity and aldosterone biosynthesis, two actions that should contribute to the beneficial effect of the drug in the treatment of hypertension. PMID- 9864259 TI - Delta-opioid ligands reverse alfentanil-induced respiratory depression but not antinociception. AB - Evidence suggests both opioid mu and delta receptors may participate in the regulation of respiration at different central nervous system sites. In the past, the overlapping receptor specificity of various opioid drugs has made it difficult to dissect the receptor subtype-specific activities involved in respiratory regulation. The new family of delta receptor selective agents such as cyclic[D-Pen2, 5]enkephalin, deltorphins, (+)-4-((alpha-R)-alpha-((2S,5R)-4-allyl 2, 5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N,N-diethylbenzamide, naltrindole and H-Tyr-Tic(psi)[CH2NH]Phe-Phe-OH have now made it feasible to more clearly define the role of delta receptors in respiratory control. In a series of experiments we observed that systemic infusion of rats with the highly mu receptor-specific opioid alfentanil induced antinociception and hypercapnia, and both of these effects were antagonized by the mu antagonist D-Phe-Cys-Tyr-Orn-Thr Pen-Thr-NH2. However, peripheral administration of the delta receptor antagonist naltrindole reverses the hypercapnia but not the antinociceptive activity of alfentanil. This differential effect of naltrindole on antinociception and hypercapnia could also be produced with the delta agonist (+)-4-((alpha-R)-alpha ((2S,5R)-4-allyl-2, 5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N,N diethylbenzamide. In addition, intracerebroventricular delivery of a number of peptide delta ligands cyclic[D-Pen2,5]enkephalin, deltorpnin II and H-Tyr Tic(psi)[CH2NH]Phe-Phe-OH also produced the same differential reversal of hypercapnia without affecting antinociception. Thus, both the traditional delta agonists and antagonists are able to reverse the alfentanil-induced hypercapnia without affecting antinociception. The reversal of alfentanil-induced hypercapnia by these delta ligands was antagonized by a novel synthetic delta antagonist cis 4-(alpha-(4-((Z)-2-butenyl)-3, 5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N,N diethylbenzamide. We propose that in this experimental respiration model, the delta antagonists naltrindole and H-Tyr-Tic(psi)[CH2NH]Phe-Phe-OH behave like delta agonists with low but sufficient intrinsic activities to reverse alfentanil induced hypercapnia in rats. The results suggest that a function of the delta receptor is to modulate or counteract the respiratory depression induced by the mu receptor. PMID- 9864261 TI - Role of vasoactive intestinal polypeptide in the adaptation of intestinal smooth muscle cells to mechanical distension. AB - Distension of the small intestine can play a role in the pathogenesis of various functional intestinal disorders. This study determined the role of vasoactive intestinal polypeptide (VIP) in the adaptative response of intestinal smooth muscle to acute and chronic distension of the ileum in vivo. Several in vitro experiments were performed to identify the mechanism of receptor regulation. Distension was applied by a balloon inflated with air in the ileum either during a single episode in anesthetized or repeatedly in conscious guinea pigs. Then, muscle cells were isolated by enzymatic digestion from the distended and nondistended adjacent ileal segments. In addition, in vitro experiments were performed on freshly dispersed cells for determination of mechanisms. Control cells maximally relaxed (Cmax) at 1 microM VIP (EC50 = 50 pM) and 100 microM isoproterenol (EC50 = 7 nM). Both acute and chronic distensions triggered a right ward shift of the concentration-response curves for VIP (Cmax = 100 microM, EC50 = 10 nM). A desensitization of the relaxing effect of VIP receptors was also observed when cells were preincubated for 30 min in vitro with VIP. By contrast, the relaxing effect of isoproterenol was affected neither by in vivo distension nor by in vitro incubation with isoproterenol. Desensitization of VIP receptors was prevented by in vitro incubation of cells with VIP plus a VIP antagonist [(D P-Cl-Phe6,Leu17)VIP] and by intraluminal perfusion of the VIP antagonist during acute distention in vivo. Moreover, desensitization of VIP receptors did not occur after 30 min preincubation with either forskolin or 8-Bromo-cyclic AMP. These results indicate that mechanical distension of the ileum induces a homologous desensitization of VIP receptors on circular smooth muscle cells, which requires the occupation of its receptors by VIP. PMID- 9864262 TI - Effects of 5-hydroxytryptamine2 receptor antagonism on the behavioral activation and immediate early gene expression induced by dizocilpine. AB - The noncompetitive N-methyl-D-aspartate (NMDA) antagonists dizocilpine and phencyclidine cause behavioral changes in animals that can be blocked by antipsychotic agents, implicating NMDA receptors in the expression of schizophrenic symptoms. In the present study, we examined the effects of dizocilpine (0.1-3.0 mg/kg s.c.) on locomotor activity and on the expression of c fos and hsp-70 immediate-early genes (IEGs) in mice. Results indicate that dizocilpine increases locomotor activity and selectively increases the expression of c-fos and hsp-70 in the posterior cingulate cortex. Haloperidol (0.01-0.1 mg/kg) and clozapine (0.6-1.25 mg/kg) block both the locomotor response and the increased IEG immunoreactivity induced by dizocilpine (0.6 mg/kg). The 5-HT2 antagonists ritanserin (0.06-0.25 mg/kg), ketanserin (0.03-0.12 mg/kg) and amesergide (0. 3-1.25 mg/kg) also significantly attenuated the locomotor response to dizocilpine. Haloperidol and clozapine suppressed the head weaving induced by dizocilpine, but ritanserin, as previously reported did not. Although some attenuation of the c-fos and hsp-70 immunoreactivity was seen with the 5-HT2 antagonists it was less pronounced than that induced by haloperidol or clozapine. In conclusion, 5-HT2 antagonists as well as antipsychotic compounds attenuate the locomotor response to dizocilpine in mice. Haloperidol and clozapine appear to be more effective, however, in attenuating the expression of c-fos and hsp-70 in the posterior cingulate gyrus than 5-HT2 antagonists ritanserin, ketanserin or amesergide. We thus have seen a dissociation in the capacity of compounds to alter the effects on behavior and IEG expression after dizocilpine administration. PMID- 9864263 TI - Differences between the antinociceptive effects of the cholinergic channel activators A-85380 and (+/-)-epibatidine in rats. AB - (+/-)-Epibatidine (EPIB) and A-85380 are nicotinic acetylcholine receptor (nAChR) agonists that bind to the agonist ([3H]cytisine) binding site with 40 to 50 pM affinity but have different affinities in nAChR subtype selective functional receptor assays. In vivo EPIB was more (23-fold) potent than A-85380 in reducing open field activity and more (12-fold) potent in reducing nociception in the formalin test of persistent chemical pain. In the rat hot box test of thermal acute pain, both compounds produced antinociception, as indicated by an increase in the paw withdrawal latency, however EPIB was a approximately 33-fold more potent than A-85380 (ED50 = 0.004 and 0.11 micromol/kg, i.p., respectively). The systemic effects of both nAChR agonists were blocked by central (i.c.v.) administration of the nAChR antagonist chlorisondamine suggesting a central site of action for these compounds. Injections of EPIB (0.0013 to 0.013 nmol) and A 85380 (0.013 to 0.13 nmol) directly into the nucleus raphe magnus (NRM) were also effective in the hot box and could be blocked by coadministration of the nAChR antagonists chlorisondamine (0.23 nmol) or mecamylamine (0.8 nmol). The NRM was found to be critical for the antinociceptive effects of systemic EPIB but not for A-85380 in that NRM injections of either mecamylamine (0.8 nmol) or lidocaine (74 nmol) blocked the antinociceptive effects of systemic (i.p.) EPIB but not those of A-85380. These results suggest that A-85380 may act at multiple sites both within and outside the NRM, whereas EPIB acts largely via descending inhibitory pathways arising from the NRM. PMID- 9864264 TI - Antitumor necrosis factor therapy in rat chronic granulomatous colitis: critical dose-timing effects on outcome. AB - Inhibition of tumor necrosis fact (TNFalpha) is of potential benefit in the treatment of chronic inflammatory conditions. However, TNFalpha plays an important role in host defenses against infection, and blocking TNFalpha production may also have adverse effects. We tested the efficacy and safety of anti-TNFalpha therapy in experimental colitis induced by trinitrobenzenesulfonic acid. We cultured colonic wall specimens for bacterial growth and measured native TNFalpha protein synthesis in colonic tissue at days 0, 1, 4, 10 and 18 after induction of colitis. Anti-TNFalpha therapy (monoclonal g1 immunoglobulin, 15 mg/kg i.p., every third day) was started on either day 4 or day 10 after induction of colitis. On day 18, we measured the release of inflammatory mediators and scored colonic lesions. In acute lesions, several species of the common flora were grown, including Streptococcus, Staphylococcus, Bacteroides, clostridia and enterobacteria. In chronic lesions, only enterobacteria, clostridia and lactobacilli were isolated. TNFalpha production by inflamed colonic tissue was increased in both acute and chronic lesions. Anti-TNFalpha therapy induced a significant decrease in the release of inflammatory mediators and histopathological remission when treatment started on day 10. However, anti TNFalpha therapy increased eicosanoid release and lesion scores when treatment started on day 4. In conclusion, acute colonic lesions showed polymicrobial infection. Anti-TNFalpha therapy induced remission of chronic intestinal inflammation, but early treatment did not prove effective. PMID- 9864265 TI - OPC-41061, a highly potent human vasopressin V2-receptor antagonist: pharmacological profile and aquaretic effect by single and multiple oral dosing in rats. AB - The pharmacological profile and the acute and chronic aquaretic effects of OPC 41061, a novel nonpeptide human arginine vasopressin (AVP) V2-receptor antagonist, were respectively characterized in HeLa cells expressing cloned human AVP receptors and in conscious male rats. OPC-41061 antagonized [3H]-AVP binding to human V2-receptors (Ki = 0.43 +/- 0.06 nM) more potently than AVP (Ki = 0. 78 +/- 0.08 nM) or OPC-31260 (Ki = 9.42 +/- 0.90 nM). OPC-41061 also inhibited [3H] AVP binding to human V1a-receptors (Ki = 12.3 +/- 0.8 nM) but not to human V1b receptors, indicating that OPC-41061 was 29 times more selective for V2-receptors than for V1a-receptors. OPC-41061 inhibited cAMP production induced by AVP with no intrinsic agonist activity. In rats, OPC-41061 inhibited [3H]-AVP binding to V1a-receptors (Ki = 325 +/- 41 nM) and V2-receptors (Ki = 1.33 +/- 0. 30 nM), showing higher receptor selectivity (V1a/V2 = 244) than with human receptors. A single oral administration of OPC-41061 in rats clearly produced dose-dependent aquaresis. In treatment by multiple OPC-41061 dosing for 28 days at 1 and 10 mg/kg p.o. in rats, significant aquaretic effects were seen throughout the study period. As the result of aquaresis, hemoconcentration was seen at 4 hr postdosing although, no differences were seen in serum osmolality, sodium, creatinine and urea nitrogen concentrations at 24 hr postdosing. Furthermore, there was no difference in serum AVP concentration, pituitary AVP content or the number and affinity of AVP receptors in the kidney and liver at trough throughout the study period. These results demonstrate that OPC-41061 is a highly potent human AVP V2 receptor antagonist and produces clear aquaresis after single and multiple dosing, suggesting the usefulness in the treatment of various water retaining states. PMID- 9864266 TI - Role of lipid peroxidation in dapsone-induced hemolytic anemia. AB - Dapsone hydroxylamine (DDS-NOH) is a direct-acting hemolytic agent responsible for dapsone-induced hemolytic anemia in the rat. The hemolytic activity of DDS NOH is associated with the formation of disulfide-linked hemoglobin adducts on membrane skeletal proteins. We have postulated that this membrane protein "damage" is a consequence of DDS-NOH-induced oxidative stress within the red cell and that it serves as the trigger for premature removal of injured but intact red cells from the circulation by splenic macrophages. Oxidative stress has also been associated with the induction of lipid peroxidation, and it is possible that direct damage to the lipoidal membrane may play a role in the premature sequestration of the damaged cells in the spleen. To investigate this possibility, rat and human red cells were incubated with hemolytic concentrations of DDS-NOH and examined for evidence of lipid peroxidation using two independent assays: thiobarbituric acid-reactive substances formation and cis-paranaric acid degradation. Phenylhydrazine, which is known to induce lipid peroxidation in red cells, was used as a positive control. The extent of thiobarbituric acid-reactive substances formation and cis-paranaric acid degradation in DDS-NOH-treated rat and human red cells was not significantly different from that in control cells. In contrast, thiobarbituric acid-reactive substances formation and cis-paranaric acid degradation were significantly increased in red cells treated with hemolytic concentrations of the positive control, phenylhydrazine. These data suggest that lipid peroxidation is not involved in the mechanism underlying dapsone-induced hemolytic anemia. PMID- 9864267 TI - The antiestrogen tamoxifen blocks the delayed rectifier potassium current, IKr, in rabbit ventricular myocytes. AB - Tamoxifen is an antiestrogen drug commonly used to treat breast cancer and has been shown to cause prolongation of the electrocardiographic QT interval in humans. Because QT prolongation could influence cardiac arrhythmias, we sought to determine the electrophysiologic mechanism(s) underlying the tamoxifen action. The whole-cell patch-clamp technique was used to study the effect of tamoxifen on the delayed rectifier (IKr), the inward rectifier (IK1), the transient outward current (Ito), and the inward L-type calcium current (ICa) in rabbit ventricular myocytes. By switching to the current-clamp mode, the effect of tamoxifen on action potential duration (APD) was also studied. Tamoxifen blocked IKr in a time , concentration- and voltage-dependent fashion. IKr tail currents were completely blocked by 10 micromol/l tamoxifen with no recovery after 15 min of washout. At +50 mV, tamoxifen 1 and 3.3 micromol/l blocked IKr by 39.5 +/- 1.7% (P <.01) and 84.8 +/- 1.3% (P <.01) respectively, while no significant block of IK1 or Ito was observed. Significant block of ICa by tamoxifen was also observed at concentrations greater than 1 micromol/l, with almost complete inhibition at 10 micromol/l. Tamoxifen showed no significant effect on APD at concentrations up to 3.3 micromol/l. We conclude that tamoxifen potently blocks both IKr and ICa at clinically relevant concentrations. The observed QT prolongation by tamoxifen in humans may be a result of its predominant effect on IKr. Inhibition of IKr, in conjunction with other QT-prolonging factors in patients could increase their risk of developing torsades de pointes-type cardiac arrhythmias. PMID- 9864268 TI - Dual activation and inhibition of adenylyl cyclase by cannabinoid receptor agonists: evidence for agonist-specific trafficking of intracellular responses. AB - Cannabinoid receptors couple to both Gs and Gi proteins and can consequently stimulate or inhibit the formation of cAMP. To test whether there is specificity among cannabinoid receptor agonists in activating Gs- or Gi-coupled pathways, the potency and intrinsic activity of various cannabinoid receptor ligands in stimulating or inhibiting cAMP accumulation were quantified. The rank order of potencies of cannabinoid receptor agonists in increasing or inhibiting forskolin stimulated cAMP accumulation, in CHO cells expressing hCB1 receptors, was identical (HU-210 > CP-55,940 > THC > WIN-55212-2 > anandamide). However, the activities of these agonists were different in the two assays with anandamide and CP-55,940 being markedly less efficacious in stimulating the accumulation of cAMP than in inhibiting its formation. Studies examining the effects of forskolin on cannabinoid receptor mediated stimulation of adenyly cyclase also revealed differences among agonists in as much as forskolin enhanced the potency of HU-210 and CP-55,940 by approximately 100-fold but, by contrast, had no effect on the potency of WIN-55212-2 or anandamide. Taken together these findings demonstrate marked differences among cannabinoid receptor agonists in their activation of intracellular transduction pathways. This provides support for the emerging concept of agonist-specific trafficking of cellular responses and further suggests strategies for developing receptor agonists with increased therapeutic utility. PMID- 9864269 TI - Pardaxin, a new pharmacological tool to stimulate the arachidonic acid cascade in PC12 cells. AB - The effect of Pardaxin, a neurotoxin that induces neurotransmitter release from neurons, on the arachidonic acid (AA) cascade was studied in PC12 cells. Both native and the synthetic Pardaxin selectively stimulated phospholipase A2 (PLA2) activity (measured by [3H]AA release) in the presence as well as in the absence of extracellular calcium. Pardaxin-stimulated PLA2 activity was also evident in the increased formation of lysophosphatidylcholine. Pardaxin analogs, lacking the alpha-helical structure that is essential for insertion into the plasma membrane, were ineffective in stimulating the AA cascade in PC12 cells. Pardaxin stimulation of PLA2 was markedly inhibited by the nonselective PLA2 inhibitors bromophenacyl bromide and mepacrine, by methyl arachidonyl fluorophosphonate, a dual inhibitor of calcium-dependent cytosolic PLA2 and the calcium-independent PLA2 and by bromoenol lactone[(E)-6-(bromoethylene)tetrahydro-3-(1-naphthalenyl 2H-pyran -2- one], a highly specific inhibitor of calcium-independent PLA2. After Pardaxin treatment, there was increased release of AA metabolites produced by the cyclooxygenase pathway as expressed in an 8-fold increase of PGE2 release. The release of other eicosanoids, such as 6-keto-PGF1alpha and thromboxane B2, was also augmented. Pardaxin-induced PGE2 release was observed in calcium-free medium and in the absence of any increase in cytosolic calcium. Dexamethasone partially inhibited Pardaxin-induced PGE2 release. This effect was reversed by the type II corticosteroid receptor antagonist RU-38486. Our results indicate that Pardaxin stimulates release of AA and eicosanoids, independently of calcium, and suggest that calcium-independent PLA2 plays an important role in Pardaxin stimulation of the AA cascade. PMID- 9864270 TI - Polyol pathway hyperactivity is closely related to carnitine deficiency in the pathogenesis of diabetic neuropathy of streptozotocin-diabetic rats. AB - To investigate the relationship between polyol pathway hyperactivity and altered carnitine metabolism in the pathogenesis of diabetic neuropathy, the effects of an aldose reductase inhibitor, [5-(3-thienyl) tetrazol-1-yl]acetic acid (TAT), and a carnitine analog, acetyl-L-carnitine (ALC), on neural functions and biochemistry and hemodynamic factors were compared in streptozotocin-diabetic rats. Significantly delayed motor nerve conduction velocity, decreased R-R interval variation, reduced sciatic nerve blood flow and decreased erythrocyte 2, 3-diphosphoglycerate concentrations in diabetic rats were all ameliorated by treatment with TAT (administered with rat chow containing 0.05% TAT, approximately 50 mg/kg/day) or ALC (by gavage, 300 mg/kg/day) for 4 weeks. Platelet hyperaggregation activity in diabetic rats was diminished by TAT but not by ALC. TAT decreased sorbitol accumulation and prevented not only myo-inositol depletion but also free-carnitine deficiency in diabetic nerves. On the other hand, ALC also increased the myo-inositol as well as the free-carnitine content without affecting the sorbitol content. These observations suggest that there is a close relationship between increased polyol pathway activity and carnitine deficiency in the development of diabetic neuropathy and that an aldose reductase inhibitor, TAT, and a carnitine analog, ALC, have therapeutic potential for the treatment of diabetic neuropathy. PMID- 9864271 TI - Hypoxia stimulates the synthesis of cytochrome P450-derived inflammatory eicosanoids in rabbit corneal epithelium. AB - The corneal epithelium metabolizes arachidonic acid by a cytochrome P450-(CYP) mediated pathway to 12(R)hydroxy-5,8,10,14-eicosatrienoic acid [12(R)-HETE] and 12(R)hydroxy-5,8,14-eicosatrienoic acid [12(R)-HETrE]. Both metabolites possess potent inflammatory properties with 12(R)-HETrE being a powerful angiogenic factor and assume the role of inflammatory mediators in hypoxia- and chemical induced injury in the cornea, in vivo. We developed an in vitro model of corneal organ culture to characterize the biochemical and molecular events involved in the increased synthesis of these metabolites. These cultured corneas exhibit epithelial cytochrome P450 CYP-dependent 12(R)-HETE and 12(R)-HETrE synthesis as indicated by chiral analysis and by the ability of CYP enzyme inhibitors to repress their synthesis. Hypoxia greatly and selectively stimulated the synthesis of 12(R)-HETE (7-fold over control normoxic conditions) and 12(R)-HETrE. The bacterial endotoxin, lipopolysaccharide, also increased the synthesis of these eicosanoids, substantiating the notion that this activity may function as an inflammatory pathway. These metabolites were detected in the culture medium by gas chromatography/mass spectroscopy (GC/MS) analysis and their levels significantly increased in hypoxia-treated corneas, further indicating their endogenous formation in response to injury. This in vitro model provides an excellent preparation for studying factors regulating the synthesis of these inflammatory eicosanoids and for isolating, identifying and characterizing the CYP protein responsible for their synthesis. PMID- 9864272 TI - Effects of P-glycoprotein modulators on etoposide elimination and central nervous system distribution. AB - In this study, P-glycoprotein modulator effects on pharmacokinetics and central nervous system distribution of the chemotherapeutic agent etoposide were evaluated. The multidrug resistance transporter P-glycoprotein is expressed in normal tissues, and its physiological function is thought to be an excretory and/or protective one. To examine this further, we evaluated etoposide under steady-state and bolus dose conditions. In microdialysis infusion studies, etoposide 15 mg/kg/hr was administered to 12 rats. Rats received sodium cyanide (1 or 100 mM), trifluoperazine (30 mM) or cyclosporine (4.14 mM) via microdialysis probe at 3.5 hr after etoposide infusion initiation. High-dose sodium cyanide (100 mM) increased the etoposide BBR,corr from 0.09 +/- 0.03 to 0.85 +/- 0.35. Similarly, trifluoperazine significantly increased the BBR,corr (0.05 +/- 0.02 vs. 1.30 +/- 0.43), whereas cyclosporine had no effect. In bolus studies, etoposide (10-12 mg/kg) was given alone or concomitant to cyclosporine (5 mg/kg) or tamoxifen (13.5 mg/kg). Control etoposide total systemic clearance (ml/min/kg) was 29.3 +/- 13.0 vs. 16.0 +/- 1.9 and 22.6 +/- 5.3 for cyclosporine and tamoxifen treatments, respectively. Etoposide nonrenal clearance (ml/min/kg) values for cyclosporine (12.0 +/- 1.6) and tamoxifen (18.1 +/- 3.6) treatments was also decreased from controls (23.5 +/- 10.5). Etoposide renal clearance (ml/min/kg) values (5.7 +/- 2.5) were not significantly different from cyclosporine (4.0 +/- 0.7) or tamoxifen (4.6 +/- 1.7) treatments, respectively. In this study, the ability of sodium cyanide and trifluoperazine to alter etoposide BBR,corr, demonstrated that etoposide distribution into brain is partly controlled by an active transport process. Similarly, the results indicate cyclosporine inhibits etoposide transport at the canalicular membrane and/or etoposide P-450 metabolism. PMID- 9864273 TI - Analysis of 3-(4-hydroxy, 2-Methoxybenzylidene)anabaseine selectivity and activity at human and rat alpha-7 nicotinic receptors. AB - 3-(2,4-dimethoxybenzylidene)anabaseine (GTS-21) is a selective partial agonist for rat alpha-7 nicotine receptors with reportedly much lower efficacy for human alpha-7 receptors. Because this drug improves memory-related performance in nonhuman primates, and is presently in a clinical trial for Alzheimer's disease, we investigated the potential effects of its primary human metabolite, 3-(4 hydroxy, 2-methoxy-benzylidene)anabaseine) on human as well as rat nicotinic acetylcholine receptor. 4OH-GTS-21 exhibited a similar level of efficacy for both rat and human alpha-7 receptors expressed in Xenopus oocytes. It displaced high affinity [125I]alpha-bungarotoxin binding to human SK-N-SH cell-membranes (Ki 0.17 microM) and rat PC12 cell-membranes (Ki 0.45 microM). GTS-21 also displaced [125I]alpha-bungarotoxin binding to PC12 cell membranes with high potency (Ki 0.31 microM), but was much less potent in this regard in SK-N-SH cells (23 microM). 4OH-GTS-21 produced less residual inhibition of either the human or rat AChR subtypes than GTS-21 did. To compare the neuroprotective efficacies of GTS 21 and 4OH-GTS-21 in both species, an amyloid-toxicity model (Abeta 25-35) was used. 4OH-GTS-21 was protective in both human and rat cell lines, although GTS-21 was effective only in the latter. These studies suggest that the efficacy of GTS 21 in primates may depend on a pro-drug function. PMID- 9864274 TI - Evidence for an endogenous cAMP-adenosine pathway in the rat kidney. AB - In the rat kidney, exogenous adenosine-3'-5'-monophosphate (cAMP) is converted to adenosine via the metabolism of cAMP to adenosine-5'-monophosphate by phosphodiesterase and adenosine-5'-monophosphate to adenosine by 5'-nucleotidase. Our purpose was to investigate whether in the rat kidney adenosine is synthesized from endogenous cAMP via the same pathway. Rat kidneys were perfused with Tyrode's solution, and stabilized for 3 hr to minimize basal renal purine secretion. In control experiments (n = 6), the renal venous secretion rate of adenosine, inosine, hypoxanthine and Sigmapurines (adenosine + inosine + hypoxanthine) did not change over the two 10-min experimental periods. In contrast, the beta adrenoceptor agonist (+/-)-isoproterenol (1 and 10 microM added to the perfusate) caused a significant (1-factor analysis of variance with repeated measures; n = 31) increase in the renal venous secretion of adenosine (P <.0001), inosine (P <.0007), hypoxanthine (P <.0007) and Sigmapurines (P <.0001) as measured by high-performance liquid chromatography with ultraviolet detection. The Sigmapurines was the most discriminating index of isoproterenol-induced changes in purine release, and the renal venous secretion of Sigmapurines was significantly (2-factor analysis of variance with repeated measures) attenuated by inhibition of beta adrenoceptors with propranolol (.1 microM, n = 6; P <.05), phosphodiesterase with 3-isobutyl-1-methylxanthine (1 mM, n = 5; P <.002) and 5' nucleotidase with alpha, beta-methyleneadenosine-5'-diphosphate (0.1 mM, n = 5; P <.03). Our data indicate that activation of beta adrenoceptors increases purine biosynthesis in the rat kidney via a mechanism that involves phosphodiesterase and 5'-nucleotidase. These results support the existence of an endogenous cAMP adenosine pathway in the rat kidney. PMID- 9864275 TI - Aldosterone receptor blockade inhibits increased furosemide-sensitive sodium reabsorption in rats with liver cirrhosis. AB - We examined the role of chronic aldosterone receptor blockade on the altered furosemide-sensitive sodium reabsorption in rats with liver cirrhosis induced by common bile duct ligation. CBL and sham-operated control animals were treated with the aldosterone receptor antagonist canrenoate (20 mg/day i.v.) for 4 weeks. Untreated CBL and sham-CBL served as control groups. The plasma concentration of aldosterone was within the normal range in all groups. Sodium balance studies showed that aldosterone receptor blockade prevented sodium retention in cirrhotic rats. Clearance studies showed that the glomerular filtration rate was unchanged, whereas the renal plasma flow was increased in CBL rats. A test dose of furosemide (7.5 mg/kg b.wt. i.v.) produced significantly greater diuretic (+59%) and natriuretic (+56%) responses in CBL rats than in sham-operated controls. The urinary furosemide excretion rate (UFURV) reflects delivery of furosemide to the thick ascending limb. When the natriuresis was expressed relative to UFURV (i.e., the natriuretic efficiency), we found that natriuretic efficiency of furosemide was significantly increased in untreated CBL rats (+59%). However, the natriuretic efficiency of furosemide was normalized in CBL rats treated with canrenoate. The urinary excretion of furosemide was unchanged in untreated CBL rats, but it was significantly increased in cirrhotic rats treated with canrenoate (+43%). This suggests that in CBL rats, chronic canrenoate treatment increases the renal elimination of furosemide as a consequence of reduced metabolism. These data suggest that chronic aldosterone receptor blockade with canrenoate prevents sodium retention in cirrhotic rats partly by inhibition of increased sodium reabsorption in the thick ascending limb. PMID- 9864276 TI - Spinal morphine/clonidine antinociceptive synergism is regulated by protein kinase C, but not protein kinase A activity. AB - When coadministered spinally, morphine and clonidine interact synergistically to produce antinociception. The mechanism for the synergism is unknown, but may depend on intracellular messenger systems. Agents that alter the activities of protein kinases alter antinociception produced by opioids, but their effects on clonidine-induced antinociception or the morphine/clonidine interaction are not known. In these studies, mice were pretreated intrathecally with inhibitors or activators of protein kinase C and cyclic AMP-dependent protein kinase (protein kinase A). Antinociceptive responses to intrathecally administered morphine, clonidine and morphine/clonidine combinations were then measured in the radiant heat tail flick test. Inhibition of protein kinase C activity with chelerythrine or calphostin C changed the morphine/clonidine interaction from synergistic to additive. Inhibition of protein kinase A activity with H-89 did not alter the morphine/clonidine interaction, it remained synergistic. Stimulation of protein kinase C activity with phorbol 12,13-dibutyrate attenuated morphine antinociception, but did not alter the synergistic interaction. Increasing spinal cyclic AMP concentrations with either forskolin or rolipram attenuated the antinociception produced by separately administered morphine and clonidine, but had no effect on the morphine/clonidine interaction. These results suggest that protein kinase C activity may regulate the interaction between spinal opioid and alpha-2 receptors, stimulated by morphine and clonidine. PMID- 9864277 TI - Substrate specificity of organic cation/H+ exchange in avian renal brush-border membranes. AB - The substrate specificity of the avian renal organic cation exchanger was examined in isolated renal brush-border membrane vesicles. Endobiotic and xenobiotic organic cations (OCs) were tested at a concentration of 100 microM for cis-inhibition of 14C-tetraethylammonium (TEA)/H+ exchange and at 1 mM for trans stimulation of 14C-TEA efflux. The xenobiotic cations amiloride, cimetidine, mepiperphenidol, procainamide, quinidine, quinine, and ranitidine cis-inhibited TEA uptake >/= 80%; isoproterenol and unlabeled TEA inhibited uptake at least 30%. In contrast, the endogenous cations acetylcholine, choline, and guanidine did not inhibit TEA uptake; however, epinephrine, N1-methylnicotinamide, serotonin, and thiamine inhibited uptake as much as 60%. Each endogenous cation, except thiamine, trans-stimulated TEA efflux, and xenobiotic cations, excluding isoproterenol and TEA, trans-inhibited TEA efflux. The data suggest that the avian renal tubule luminal OC exchanger has greater affinity for xenobiotic cations than for endobiotic cations, but greater transport capacity for endobiotics than for xenobiotics. PMID- 9864278 TI - The role of histamine H1, H2 and H3 receptors on enteric ascending synaptic transmission in the guinea pig ileum. AB - The role of histamine H1-, H2- and H3-receptors was studied on neural transmission in ascending excitatory pathways of the guinea pig ileum. A two compartment (oral and anal compartments) bath was used: ascending neural pathways were activated by electrical stimulation in the anal compartment and the resulting contraction of the circular muscle in the oral compartment was recorded. Drugs were applied in the anal compartment and each agonist was evaluated in the presence of the antagonists of the other two receptors. In the presence of cimetidine (10 microM) and thioperamide (1 microM), histamine (0.03-3 microM) depressed the nerve-mediated contractions (5-70% inhibition, P <.05-.01). The inhibitory effect of histamine was antagonized by mepyramine. At the higher concentrations (10 and 30 microM), histamine elicited contractions of the circular muscle in the oral compartment, and these were abolished by mepyramine (1 microM) and tetrodotoxin (0.6 microM). The H2 agonists dimaprit (30 and 100 microM) and amphamine (0.1-300 microM) produced small contractions of the circular muscle in the oral compartment. These contractile responses were abolished by tetrodotoxin (0.6 microM) and cimetidine (10 microM). The H3 agonist R-alpha-methylhistamine (0.001-1 microM) inhibited (2-58%, P <.05) the nerve mediated contractions. This inhibitory effect was antagonized by the H3 antagonist thioperamide. These results indicate that 1) histamine, acting at H1 receptors, at lower concentrations depresses synaptic transmission, although at higher concentrations activates the enteric excitatory ascending pathway; 2) activation of H2 receptors by H2 agonists stimulates the enteric excitatory ascending pathways and 3) activation of H3 receptors inhibits synaptic transmission. PMID- 9864279 TI - Suppression of nicotine intake during ad libitum cigarette smoking by high-dose transdermal nicotine. AB - Nicotine replacement therapy is believed to facilitate smoking cessation both by relieving withdrawal symptoms and by reducing the psychological reward from smoking. The latter might occur via down-regulation of nicotine receptors in the brain, which might require high levels of nicotine exposure. Our study examined the hypothesis that transdermal nicotine, dosed up to three times the doses currently recommended for smoking cessation, would suppress nicotine intake from ad libitum smoking in a dose-dependent manner. Eleven volunteers with no desire to quit smoking received placebo or 21, 42, and 63 mg/day transdermal nicotine, with and without cigarette smoking, in a blinded crossover study. Cigarette smoking was permitted as desired. Transdermal nicotine suppressed nicotine intake from cigarette smoking by 3%, 10% and 40% on average in the 21, 42 and 63 mg/day conditions. The number of cigarettes smoked per day declined from an average of 17.2 to 12.7 and the intake of nicotine per cigarette declined from 2.5 to 1.6 mg, comparing placebo and 63 mg nicotine conditions. Our study results suggest that high-dose transdermal nicotine has the potential to substantially suppress the intake of tobacco smoke and could be a useful strategy for smoking cessation therapy or for reducing the harm caused by smoking. PMID- 9864280 TI - Modulation of the tumor disposition of vinca alkaloids by PSC 833 in vitro and in vivo. AB - PSC 833, a nonimmunosuppressive cyclosporin, is able to inhibit the efflux of antitumor drugs mediated by P-glycoprotein (P-gp). The purpose of the present study is to compare the effect of PSC 833 on the tumor disposition of [3H]vincristine ([3H]VCR) and [3H]vinblastine ([3H]VBL) in in vitro and in vivo experiments from a pharmacokinetic point of view. In in vitro experiments, the effect of PSC 833 was investigated on the cellular uptake of [3H]VCR and [3H]VBL by HCT-15 and COLO 205, human colorectal tumor cell lines with extensive and minimal expression of P-gp, respectively. PSC 833 (2 microM) increased the cellular uptake of [3H]VCR and [3H]VBL by HCT-15 cells, but not that by COLO 205 cells, 8- and 6-fold, respectively, without affecting the initial influx rates. In addition, 2 microM PSC 833 reduced the efflux of [3H]VCR from HCT-15 cells to a level comparable with that from COLO 205 cells. Furthermore, the effect of PSC 833 on the tumor disposition of intravenously administered [3H]VCR and [3H]VBL was studied in tumor inoculated mice. Infusion of PSC 833 (10 microg/hr/mouse) increased the HCT-15 tumor disposition of [3H]VBL and [3H]VCR in vivo to a level comparable with that observed in vitro. These findings demonstrate that PSC 833 enhances the tumor disposition of vinca alkaloids by inhibition of P-gp-mediated efflux not only in vitro but also in vivo in a solid tumor model. PMID- 9864281 TI - Differential contribution of R and S isomers in ketoprofen anti-inflammatory activity: role of cytokine modulation. AB - Among nonsteroidal anti-inflammatory drugs (NSAIDs), 2-arylpropionic acids exist as a racemic mixture of its enantiomeric forms, with S-enantiomers primarily responsible for inhibition of prostaglandin synthesis and of inflammatory events. The aim of this study was to compare the anti-inflammatory effects of R- and S ketoprofen in vitro and in vivo. S-Ketoprofen efficiently inhibited carrageenan induced edema formation, but it could also amplify the LPS-induced production of the inflammatory cytokines tumor necrosis factor (TNF) and interleukin-1 (IL-1), in close correlation with its ability to inhibit prostaglandin synthesis. Because these inflammatory cytokines are among the factors involved in carrageenan induced inflammation and also are possibly involved in gastric damage, enhanced cytokine production could partially mask the analgesic effect of S-ketoprofen, and it can be associated with the clinical evidence of its gastric toxicity. On the other hand, R-ketoprofen contributes to the overall activity of the racemate, by playing the main role in ketoprofen-induced analgesia. Unlike the S-isomer, R ketoprofen did not induce a significant increase of cytokine production even at cyclooxygenase-blocking concentrations. It is concluded that the R-isomer directly contributes to the anti-inflammatory effects of ketoprofen, being more analgesic, and because it does not amplify inflammatory cytokine production. PMID- 9864282 TI - The role of CYP2C in the in vitro bioactivation of the contraceptive steroid desogestrel. AB - Desogestrel is a 3-deoxo progestogenic steroid that requires bioactivation to 3 ketodesogestrel. In these studies we have attempted to define the pathway of 3 ketodesogestrel formation and characterise the enzymes responsible for this biotransformation in vitro. Initial studies using deuterated desogestrel confirmed that desogestrel is metabolised by human liver microsomes via 3alpha hydroxy and 3beta-hydroxydesogestrel to 3-ketodesogestrel. Metabolites were analysed by radiometric high-performance liquid chromatography and were identified by liquid chromatography-mass spectrometry and by cochromatography with authentic standards. Desogestrel was metabolised by microsomes from lymphoblasts containing cDNA-expressed CYP2C9 and CYP2C19 to 3alpha hydroxydesogestrel with small amounts of 3beta-hydroxydesogestrel also being observed. The Km value for 3alpha-hydroxylation by CYP2C9 cell line microsomes was 6.5 microM and the corresponding Vmax value was 1269 pmole. mg-1. min-1. Sulfaphenazole potently inhibited 3alpha-hydroxydesogestrel formation by CYP2C9 microsomes with a Ki value of 0.91 microM. There was a significant negative correlation between 3-ketodesogestrel and CYP3A4 content/activity in a panel of human livers suggesting that the further metabolism of 3-ketodesogestrel is mediated by CYP3A4. Sulfaphenazole partially inhibited 3alpha-hydroxydesogestrel and 3-ketodesogestrel formation in human liver microsomes indicating a possible in vivo role for CYP2C9. In addition, when sulfaphenazole was combined with S mephenytoin, further inhibition of 3alpha-hydroxydesogestrel formation was observed suggesting a possible role for CYP2C19. This was confirmed in incubations with inhibitory antibodies. Whereas an anti-CYP2C9/2C19 antibody completely abolished desogestrel metabolism, anti-CYP3A4 and anti-CYP2E1 were not inhibitory. We conclude that CYP2C9 and possibly CYP2C19 and important isoforms catalysing the initial hydroxylation of desogestrel. PMID- 9864283 TI - Effects of pramipexole on contraversive rotation and functional motor impairments in 1-methyl-4-phenyl1,2,3, 6-tetrahydropyridine-induced chronic hemiparkinsonian monkeys. AB - Rotational and functional motor behavioral changes were studied in five MPTP lesioned chronic hemiparkinsonian Macaca nemestrina monkeys after i.m. pramipexole, a predominant D2 subfamily agonist. Pramipexole induced contraversive rotations in a dose-dependent manner with an optimal dose of 56 microg/kg for approximately 2 to 4 hr after injection. Three different rating scales were used to determine drug-induced functional improvement. They included a monkey parkinsonism rating scale, volitional responses to fruit presentations, and number of hand movements that appeared volitional. A dose of 56 microg/kg of pramipexole produced functional improvements on hand disability, and on a parkinsonian rating scale for monkeys in a dose-dependent manner from 32 to 100 microg/kg. These doses produced an increase in significant hand movements in the affected (contralateral) as well as in the normal (ipsilateral) hand to the side of the brain lesion compared with 5% dextrose in water vehicle control. With a dose of 100 microg/kg, the therapeutic effects of pramipexole on hand movements were less than with 56 microg/kg, due to side effects such as scratching. PMID- 9864284 TI - Antiasthmatic activity of the second-generation phosphodiesterase 4 (PDE4) inhibitor SB 207499 (Ariflo) in the guinea pig. AB - We evaluated the airway activity of the novel phosphodiesterase type 4 inhibitor SB 207499 [Ariflo; c-4-cyano-4-(3-cyclopentyloxy-4-methoxyp henyl-r-1-cyclohexane carboxylic acid)], in the guinea pig. Ovalbumin (OA)-induced contractions of guinea pig isolated tracheal strips were inhibited by SB 207499 with an EC50 of 1 microM but had little or no effect on exogenous agonist-induced contraction, which suggests that its effect on OA-induced contraction in vitro is primarily due to inhibition of mediator release from mast cells. In anesthetized guinea pigs, SB 207499 inhibited OA-induced bronchoconstriction with i.v. and p.o. ID50 values of 1.7 and 17 mg/kg, respectively. At 1, 3 and 6 hr after SB 207499 (30 mg/kg p.o.), OA-induced bronchospasm was inhibited by 92%, 70% and 58%, respectively, corresponding to elevated plasma concentrations of 1.62 +/- 0.19, 1.65 +/- 0.29 and 0. 93 +/- 0.24 microg/ml, respectively, of SB 207499. SB 207499 also inhibited house dust mite-induced bronchoconstriction (ID50 = 0.9 mg/kg i.v. and 8.9 mg/kg p.o.). In contrast to its lack of bronchorelaxant activity in vitro, SB 207499 inhibited bronchospasm induced by i.v. leukotriene D4 (LTD4) [ID50 = 3 mg/kg i.v.]. The bronchorelaxant effect of i.v.-administered SB 207499 was at least additive with that of salbutamol in reversing infused histamine enhanced airway tone, but it did not alter base line or enhance salbutamol induced cardiovascular effects. In conscious guinea pigs, SB 207499 (10 or 30 mg/kg p.o.), 1 hr before antigen or LTD4 challenge, markedly reduced bronchospasm and subsequent eosinophil influx as measured by bronchoalveolar lavage 24 hr after provocation. SB 207499 administered after OA or LTD4 challenge also reduced airway eosinophilia measured at 24 hr after OA challenge or 96 hr after LTD4 challenge. These results, coupled with the broad anti-inflammatory activity of SB 207499 previously described (Barnett et al., 1998), suggest that SB 207499 will be useful in the treatment of asthma and other inflammatory disorders. PMID- 9864285 TI - KN-93, an inhibitor of multifunctional Ca++/calmodulin-dependent protein kinase, decreases early afterdepolarizations in rabbit heart. AB - The multifunctional Ca++/calmodulin-dependent protein kinase II (CaM kinase) mediates Ca++-induced augmentation of L-type Ca++ current (ICa); therefore it may act as a proarrhythmic signaling molecule during early afterdepolarizations (EADs) due to ICa. To investigate the hypothesis that ICa-dependent EADs are favored by CaM kinase activation EADs were induced with clofilium in isolated rabbit hearts. All EADs were rapidly terminated with ICa antagonists. Hearts were pretreated with the CaM kinase inhibitor KN-93 or the inactive analog KN-92 (0.5 microM) for 10 min before clofilium exposure. EADs were significantly suppressed by KN-93 (EADs present in 4/10 hearts) compared to KN-92 (EADs present in 10/11 hearts) (P =.024). There were no significant differences in parameters favoring EADs such as monophasic action potential duration or heart rate in KN-93- or KN 92-treated hearts. CaM kinase activity in situ increased 37% in hearts with EADs compared to hearts without EADs (P =.015). This increase in CaM kinase activity was prevented by pretreatment with KN-93. In vitro, KN-93 potently inhibited rabbit myocardial CaM kinase activity (calculated Ki 100 microM). The actions of KN-93 and KN-92 on ICa and other repolarizing K+ currents did not explain preferential EAD suppression by KN-93. These data show a novel association between CaM kinase activation and EADs and are consistent with the hypothesis that the ICa and CaM kinase activation both contribute to EADs in this model. PMID- 9864286 TI - Modulatory effect of environmental stimuli on the susceptibility to amphetamine sensitization: a dose-effect study in rats. AB - In previous studies the repeated administration of 0.5 to 1.0 mg/kg of amphetamine i.v. failed to induce psychomotor sensitization if the drug was administered to animals living in the test environment (at home). The same doses did induce sensitization if animals were transported to the test environment for each drug treatment. The purpose of the present experiment was to determine the extent to which this effect of environment is dose dependent. Rats either lived in test cages or were transported from the animal colony to test cages where they received an i.v. infusion of one of five doses of amphetamine (0.125, 0.5, 1.0, 4.0 or 8.0 mg/kg) or saline each day for 5 consecutive days. Rotational behavior was used as an index of psychomotor activation. After a 6-day drug-free period all animals were challenged with 0.5 mg/kg of amphetamine to determine the pretreatment dose necessary to induce sensitization. The effect of the drug treatment environment was to shift the dose-effect curve for the induction of sensitization, such that significantly lower doses were necessary to induce sensitization when amphetamine was given in a novel environment. With high doses, however, sensitization occurred regardless of environmental condition. It is concluded that the circumstances surrounding drug administration can powerfully modulate the ability of psychostimulants to induce sensitization, but this effect is dose dependent. PMID- 9864287 TI - Magnesium-dependent inhibition of N-methyl-D-aspartate receptor-mediated synaptic transmission by ethanol. AB - Previous studies have indicated that ethanol (EtOH) has a relatively specific effect on excitatory synaptic transmission by inhibiting function of the N-methyl D-aspartate receptor. We have found that EtOH potently inhibits N-methyl-D aspartate-mediated synaptic currents in the basolateral amygdala, a brain region associated with actions of anxiolytic agents such as EtOH. This inhibitory effect of EtOH requires the presence of magnesium (Mg++). The dependence of the effect of EtOH on the presence of Mg++ suggests a possible molecular site of the action of EtOH in the vicinity of Mg++ binding sites on the N-methyl-D-aspartate receptor-channel complex. Because EtOH consumption may result in reductions in free brain Mg++, this dynamic interaction between EtOH and Mg++ may have important implications for understanding the behavioral effects of EtOH. PMID- 9864288 TI - Persistently exaggerated startle responses in rats treated with pyridostigmine bromide. AB - Troops in the Persian Gulf War have registered complaints consistent with CNS dysfunction that emerged after returning from the Gulf. A common experience among Persian Gulf War veterans was exposure to pyridostigmine bromide (PB) for prophylaxis against nerve gas exposure. To determine whether PB causes emergent CNS dysfunction, Wistar-Kyoto (WKY) and Sprague-Dawley (SD) rats were given PB for 7 consecutive days in their drinking water. The WKY, but not the SD, rats exhibited a delayed-onset, persistently exaggerated startle response. The WKY rats exhibited exaggerated startle responses that appeared 15 days after the end of PB treatment and were still evident 22 days after the end of treatment. Both the duration and the magnitude of the exaggerated startle responses were related to the dosage of PB. The PB-treated rats exhibited normal short-term and long term habituation. However, exaggerated startle responses were related to the development of enhanced short-term sensitization. Treating the rats for a second time, 7 weeks after the end of the first PB treatment, induced an exaggerated startle response that appeared sooner and dissipated faster than was evident after the first PB treatment. Inasmuch as the WKY rat has inherently low butyrylcholinesterase activity, a scavenger for PB, these results suggest that prophylactic PB may influence CNS function in individuals with low butyrylcholinesterase activity. Elaboration of the factors that mediate enhanced sensitization in the WKY rat may provide insight into some of the complaints registered by veterans of the Persian Gulf War. PMID- 9864289 TI - Chronic l-alpha acetylmethadol in rhesus monkeys: discriminative stimulus and other behavioral measures of dependence and withdrawal. AB - This study characterized discriminative stimulus and other effects of naltrexone in rhesus monkeys treated daily with the long-acting opioid l-alpha acetylmethadol (LAAM). An initial dose-finding study assessed the rate-decreasing effects of naltrexone in three monkeys receiving LAAM daily (0.32-1.78 mg/kg); subsequently, these monkeys and a fourth received 1.0 mg/kg/12 hr of LAAM although discriminating between naltrexone and saline. Responding occurred on the saline lever after the administration of LAAM, whereas >90% drug-lever responding occurred after the administration of 0.1 mg/kg of naltrexone that also elicited signs of withdrawal. Naloxone and quadazocine, but not morphine, nalbuphine or ketamine, substituted for naltrexone. Morphine and nalbuphine shifted the naltrexone dose-effect curve to the right. Compared to precipitated withdrawal, deprivation-induced withdrawal occasioned less naltrexone-lever responding and fewer observable signs of withdrawal. Maximal naltrexone-level responding occurred 24 to 48 hr after the discontinuation of LAAM treatment; the frequency of other withdrawal signs also peaked 24 to 48 hr after the discontinuation of LAAM. Partial naltrexone-lever responding occurred for up to 10 days after discontinuation of LAAM treatment; 4 and 8 days after the discontinuation of LAAM treatment, 0.1 mg/kg of naltrexone did no further increase naltrexone-lever responding or withdrawal signs suggesting that less-then-maximal naltrexone-lever responding was not due to long-lasting effects of LAAM or its metabolites. The discriminative stimuli that are associated with LAAM deprivation might be different from the stimuli associated with either training condition. This study is the first antagonist discrimination in non-humans primates treated chronically with LAAM and the results indicate that the naltrexone stimulus is related to opioid withdrawal. PMID- 9864290 TI - Modulation of CB1 cannabinoid receptor functions after a long-term exposure to agonist or inverse agonist in the Chinese hamster ovary cell expression system. AB - We have investigated the adaptive changes of the human central cannabinoid receptor (CB1) stably expressed in Chinese hamster ovary cells (CHO-CB1), after agonist (CP 55,940) or selective CB1 inverse agonist (SR 141716) treatment. CB1 receptor density and affinity constant as measured by binding assays with both tritiated ligands remained essentially unchanged after varying period exposure of CHO-CB1 cells (from 30 min to 72 hr) to saturating concentrations of CP 55,940 or SR 141716. However, using a C-myc-tagged version of the CB1 receptor, FACS analysis and confocal microscopy studies on CB1 expression indicated that the agonist promoted a disappearance of cell surface receptor although inverse agonist increased its cell surface density. Taken together these results suggest that 1) agonist induces internalization of the receptor into a cellular compartment that would be still accessible to both the hydrophobic ligands CP 55,940 or SR 141716; 2) inverse-agonist promotes externalization of the receptor from an intracellular preexisting pool to the cell surface. In parallel, we also investigated the associated effects of CP 55,940 and SR 141716 on CB1 receptor coupled second messengers. We showed that preexposure of cells to CP 55,940 induced a rapid desensitization of the CB1 to the agonist response. The ability of CP 55,940 to inhibit the forskolin-stimulated adenylyl cyclase and to activate the mitogen-activated protein kinase activity was dramatically reduced. By striking contrast, SR 141716 pretreatment of CHO-CB1 cells not only had no significant effect on the potency of CP 55,940 to inhibit the forskolin stimulated adenylyl cyclase but also induced a significant enhancement of the CP 55,940 ability to stimulate the mitogen-activated protein kinase activity. These results suggest that the modulation of the number of cell surface receptor could lead to functional desensitization or sensitization of the CB1 receptors. PMID- 9864291 TI - Mercaptoethylguanidine, a combined inhibitor of nitric oxide synthase and peroxynitrite scavenger, reduces trinitrobenzene sulfonic acid-induced colonic damage in rats. AB - The effect of mercaptoethylguanidine (MEG), a selective inhibitor of the inducible nitric oxide synthase and peroxynitrite scavenger, was evaluated in a rat model of colonic injury. A single intracolonic administration of trinitrobenzene sulfonic acid (TNBS, 20 mg/kg) dissolved in ethanol induced a severe colitis in male rats. Rats experienced bloody diarrhea and a significant loss of body weight. At 4 days after TNBS administration, the colon damage was characterized by areas of mucosal necrosis. Activity of myeloperoxidase, a marker of neutrophil infiltration, and levels of the 6-keto-prostaglandin F1alpha, were also markedly increased, whereas colonic ATP levels were reduced into the damaged tissue. Immunohistochemistry for the inducible nitric oxide synthase and nitrotyrosine, an index of nitrosative stress, showed an intense staining in the inflamed colon. Treatment with MEG (10 mg/kg i.v. b. i.d.) significantly reduced the appearance of diarrhea and the loss of body weight. This was associated with a remarkable amelioration of the disruption of the colonic architecture and suppression of the energetic failure, as well as a significant reduction of colonic myeloperoxidase activity and 6-keto-prostaglandin F1alpha levels. MEG also reduced the appearance of iNOS and nitrotyrosine immunoreactivity in the colon. The results of this study suggested that administration of MEG may be beneficial for the treatment of inflammatory bowel diseases. PMID- 9864292 TI - Cellular mechanisms of renal adaptation of sodium dependent sulfate cotransport to altered dietary sulfate in rats. AB - The renal transport and fractional reabsorption of inorganic sulfate is altered under conditions of sulfate deficiency or excess. The objective of this study was to examine the cellular mechanisms of adaptation of renal sodium/sulfate cotransport after varying dietary intakes of a sulfur containing amino acid, methionine. Female Lewis rats were divided into four groups and fed diets containing various concentrations of methionine (0, 0.3, 0.82 and 2.46%) for 8 days. Urinary excretion rates and renal clearance of sulfate were significantly decreased in the animals fed a 0% methionine diet or a 0.3% methionine diet, and significantly increased in the animals fed a 2.46% methionine diet when evaluated on days 4 and 7. Serum sulfate concentrations were unchanged by diet treatment in all animals. The fractional reabsorption of sulfate was significantly increased in the animals fed the 0% methionine diet and the 0.3% methionine diets, and decreased in the animals fed the 2.46% methionine diet. Increased mRNA and protein levels for the sodium/sulfate transporter (NaSi-1) were found in the kidney cortex following treatment with the 0 and 0.3% methionine diet groups. Sulfate homeostasis by renal reabsorption is maintained by an up-regulation of steady state levels of NaSi-1 mRNA and protein when the diet is low in methionine. PMID- 9864293 TI - HMN-1180, a small molecule inhibitor of neuronal nitric oxide synthase. AB - A newly synthesized isoquinolinesulfonamide, HMN-1180 (1-(5 isoquinolinylsulfonyl)-7-methylhomopiperazine), was shown to have selective inhibitory action against rat neuronal nitric oxide synthase (nNOS) with a Ki value of 5.4 microM. Kinetic analysis indicated that the inhibition was competitive with respect to L-arginine but not to calmodulin (CaM). However HMN 1180 exhibited no significant influence up to a concentration of 1 mM on activity of endothelial NOS (eNOS) and it was less active toward inducible NOS (iNOS) (IC50 > 100 microM). Moreover, nNOS bound to a HMN-1180-coupled Sepharose column, but eNOS and iNOS did not. These results suggest that inhibition of nNOS activity is due to direct binding of the compound to the L-arginine binding site of the synthase. Several HMN-1180 derivatives were synthesized and analyzed for their inhibitory actions against nNOS, eNOS and iNOS to cast light on its structure activity relationships. The potency of inhibition proved dependent on the position of methyl group in the homopiperazine molecule. HMN-1180 was also found to inhibit glutamate stimulated NO production generated by nNOS in the human neuroblastoma cell line SK-N-MC, thus indicating that it is useful tool for elucidating the physiological role of nNOS in neuronal function. PMID- 9864294 TI - Peripheral effects of opioids in a model of chronic intestinal inflammation in mice. AB - The study describes a model of chronic intestinal inflammation in mice. Inflammation was induced by the administration of one dose of croton oil (CO) (acute CO) or two doses (chronic CO) of intragastric CO, whereas controls received saline (SS); GI transit was measured with charcoal. Chronic CO induced intestinal inflammation substantiated by optical microscopy, weight loss (20%) and a 25% increase in GI transit. The ED50 values in SS animals were 1.67 +/- 0.13 mg/kg for morphine and 0.038 +/- 0.006 mg/kg for fentanyl; chronic CO significantly decreased the ED50 values to 0.16 +/- 0.03 mg/kg (morphine) and 0.006 +/- 0.0005 mg/kg (fentanyl). Thus the potency of morphine increased 10.4 times and that of fentanyl 6.3 times. The effects of enkephalin, but not those of U-50488H, were also significantly enhanced during chronic CO. The antitransit effects of p.o. loperamide increased 11.7 times during chronic CO. All effects were reversed by specific antagonists. The fraction of the active opioid receptor that mediates the antitransit effects of morphine was evaluated using beta funaltrexamine. In chronic CO, the doses of beta-funaltrexamine needed to antagonize 1 mg/kg of morphine were significantly higher than in SS and acute CO, and the ED50/KA ratio was 20 times lower. These results suggest an increase in the active concentration of mu-opioid receptors during chronic inflammation. PMID- 9864295 TI - Differential effects of N-methyl-D-aspartate receptor blockade on eticlopride induced immediate early gene expression in the medial and lateral striatum. AB - The function of striatopallidal neurons is regulated by N-methyl-D-aspartate (NMDA) and dopamine D2 receptors. Previous studies show that immediate early gene induction by D2 receptor blockade is suppressed by NMDA receptor antagonists. Because the pharmacology of NMDA receptors depends on the incorporation of different NR2 subunits and NR2 subunits show regional and cellular differences in their expression in striatum, our study examined whether different NMDA receptor antagonists would have differential effects on eticlopride-induced immediate early gene expression in striatum. Male Sprague-Dawley rats were pretreated with vehicle, CGS 19755, MK-801 or ifenprodil. Rats then received injections of eticlopride and were killed 40 min later. In situ hybridization histochemistry was used to determine the expression of c-fos and zif268 in the striatum. Eticlopride increased immediate early gene expression in striatum, with the increase generally being greater in lateral than in medial striatum. Pretreatment with each of the NMDA receptor antagonists dose-dependently decreased the expression of the immediate early genes. This suppression of eticlopride-induced gene expression was significant only in the medial-central aspect of striatum. Although there was a trend toward suppression of the gene induction in lateral striatum, it did not reach statistical significance and was not typically dose dependent. The data suggest that different types of NMDA receptor antagonists do not exert differential effects on D2 dopamine receptor-mediated function in the striatum. In addition, the data indicate that eticlopride-induced gene expression in the striatum is not uniformly dependent on NMDA receptor activation. PMID- 9864296 TI - Protective effect of FK409, a spontaneous nitric oxide releaser, on ischemic acute renal failure in rats. AB - The contribution of nitric oxide (NO) to ischemic acute renal failure (ARF) is controversial. In the present study, we investigated the effect of FK409 ((+/-) (E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexanamide ), a spontaneous NO donor, on ischemic ARF in rats. Ischemic ARF was induced by occlusion of the left renal artery and vein for 45 min followed by reperfusion, 2 weeks after contralateral nephrectomy. Renal functional parameters such as blood urea nitrogen, plasma creatinine, creatinine clearance, urine flow, urinary osmolality and fractional excretion of sodium were measured to test the effectiveness of the drug. Renal function in untreated ARF rats markedly decreased at 24 hr after reperfusion and thereafter tended to recover gradually. Intravenous bolus injection of FK409 at a dose of 1 mg/kg before the occlusion markedly attenuated the ischemic ARF-induced decreases in renal function, to the same extent as verapamil (1 mg/kg i.v.). The protective effect of FK409, at a dose of 3 mg/kg, was much more potent than that of the lower dose. Histopathological examination of the kidney of untreated ARF rats revealed severe renal damages, such as tubular necrosis, proteinaceous casts in tubuli and medullary congestion. These renal damages were significantly attenuated by treatment with FK409, at each dose given and this attenuation exceeded that seen with verapamil treatment. FK 409 administration led to a dose dependent increase in NO metabolites concentration in renal venous blood immediately after the reperfusion. These findings suggest that NO has a crucial role in the pathogenesis of ischemic ARF. Spontaneous NO donors may be clinically effective in cases of ischemic ARF. PMID- 9864298 TI - Ebselen protects mice against T cell-dependent, TNF-mediated apoptotic liver injury. AB - The seleno-organic drug ebselen (2-phenyl-1, 2-benzoisoselenazol-3(2H)-one) has glutathione peroxidase-like activity, and inhibits lipoxygenases, oxidative burst of leukocytes, nitric oxide synthases, protein kinases and leukocyte migration. This study elaborates in vivo in mice hitherto unknown immunopharmacological properties of ebselen. The compound was comparatively investigated in two different T cell-dependent hepatic hyperinflammation models and in two alternative models of receptor-activated liver apoptosis. Mice orally pretreated with ebselen were dose-dependently protected from concanavalin A (ConA)-induced liver injury. In livers from ebselen-pretreated mice exposed to ConA, the nuclear antiapoptotic transcription factor NFkappaB was upregulated. The release of the proinflammatory cytokine tumor necrosis factor-alpha (TNF) was downregulated, while the ciculating amount of the anti-inflammatory cytokine interleukin-10 (IL 10) was increased. Ebselen protected also from liver injury induced by the superantigen staphylococcal enterotoxin B in galactosamine (GalN)-sensitized mice. Furthermore, ebselen protected the liver and enhanced circulating IL-10 in GalN-sensitized mice treated with recombinant TNF, i.e., the common distal mediator of ConA and SEB-induced hepatotoxicity. The activation of apoptosis executing proteases, i.e., caspases, was blocked in livers of ebselen-treated mice following TNF receptor, but not following CD95 receptor activation. We propose a novel mechanism for the immunomodulatory properties of the drug and suggest that it might be useful in the therapy of T cell-mediated inflammatory disorders. PMID- 9864297 TI - Ibuprofen-induced changes in sulfate renal transport. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) increase sulfate renal clearance and decrease the fractional reabsorption of sulfate by the kidneys. The mechanism of this alteration of inorganic sulfate homeostasis is unknown. The objectives of this study were 1) to investigate if sulfate renal transport is altered in isolated membrane vesicles after pretreatment of animals in vivo with ibuprofen (IBU), and 2) to determine the cellular mechanism of changes in sulfate renal transport. Female Lewis rats received IBU at a i.v. dose of 27 mg/kg followed by an infusion of 33 microg/min for 4 hr. Sulfate transport was studied using brush border (BBM) and basolateral membrane (BLM) vesicles isolated from rat kidney cortex. The Vmax for the sodium-dependent sulfate cotransport (NaSi-1) in BBM was significantly lower in the IBU group compared with the control group (0.79 +/- 0.23 vs. 1.25 +/- 0.17 nmol/mg protein/10 sec, respectively; P <.05) with no change in Km. There were no significant differences between the study groups in sulfate anion exchange kinetics in BLM vesicles. NaSi-1 transporter mRNA level in kidney cortex and protein level in BBM were significantly lower in animals pretreated with IBU compared with that in control animals. There was no change in membrane fluidity of BBM and BLM isolated from IBU-treated animals as measured by the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene. These results indicate that IBU treatment alters sodium-dependent sulfate cotransport by a downregulation of mRNA and protein of NaSi-1 transporter in BBM. PMID- 9864299 TI - Transforming growth factor-beta 1 (TGF-beta1) promotes IL-2 mRNA expression through the up-regulation of NF-kappaB, AP-1 and NF-AT in EL4 cells. AB - Transforming growth factor beta1 (TGF-beta1) has been previously shown to modulate interleukin 2 (IL-2) secretion by activated T-cells. In the present studies, we determined that TGF-beta1 induced IL-2 mRNA expression in the murine T-cell line EL4, in the absence of other stimuli. IL-2 mRNA expression was significantly induced by TGF-beta1 (0.1-1 ng/ml) over a relatively narrow concentration range, which led to the induction of IL-2 secretion. Under identical condition, we examined the effect of TGF-beta1 on the activity of nuclear factor AT (NF-AT), nuclear factor kappaB (NF-kappaB), activator protein-1 (AP-1) and octamer, all of which contribute to the regulation of IL-2 gene expression. Electrophoretic mobility shift assays showed that TGF-beta1 markedly increased NF-AT, NF-kappaB and AP-1 binding to their respective cognate DNA binding sites, whereas octamer binding remained constant, as compared with untreated cells. Employing a reporter gene expression system with p(NF-kappaB)3 CAT, p(NF-AT)3-CAT and p(AP-1)3-CAT, TGF-beta1 treatment of transfected EL4 cells induced a dose-related increase in chloramphenicol acetyltransferase activity that correlated well with the DNA binding profile found in the electrophoretic mobility shift assay studies. These results show that TGF-beta1, in the absence of any additional stimuli, up-regulates the activity of key transcription factors involved in IL-2 gene expression, including NF-AT, NF-kappaB and AP-1, to help promote IL-2 mRNA expression by EL4 cells. PMID- 9864300 TI - Lipopolysaccharide activation of murine splenocytes and splenic B cells increased the expression of aryl hydrocarbon receptor and aryl hydrocarbon receptor nuclear translocator. AB - These studies characterized the profile of AhR and ARNT expression in primary splenocytes and purified splenic B cells after cellular activation with lipopolysaccharide (LPS). LPS treatment of mouse splenocytes markedly increased the magnitude of both AhR and ARNT steady state mRNA expression. AhR mRNA expression peaked at 8 hr post-LPS activation and was increased by approximately 5-fold compared with freshly isolated splenocytes (i.e., time 0). ARNT mRNA expression began to increase at 8 hr postactivation, peaked at approximately 48 hr and was increased by approximately 4-fold when compared with nonactivated splenocytes at time 0. Western blotting also demonstrated an increase in the relative magnitude of both the AhR and ARNT proteins in LPS activated splenocytes. Likewise, the presence of the AhR, ARNT and cytochrome P450IA1 (CYP1A1) proteins were also detected in purified primary splenic B cells, and the magnitude of protein expression was enhanced in LPS activated splenic B cells at 12 and 24 hr relative to time matched controls for each of these proteins. In summary, these findings suggest that on LPS activation the magnitude of AhR and ARNT mRNA and protein increases in both splenocytes and purified primary splenic B cells. Moreover, because the increase in the relative magnitude of CYP1A1 protein in response to LPS occurred in the absence of exogenous AhR ligand, these results suggest that B-cell activation is sufficient to induce AhR nuclear translocation and binding to dioxin-responsive elements in the promoter region of AhR-responsive genes. PMID- 9864301 TI - Multiple subtypes of serotonin receptors are expressed in rat sensory neurons in culture. AB - [3H]5-HT revealed the presence of serotonin receptors in cultured rat sensory neurons. [3H]5-CT binding was inhibited by cyanopindolol with an IC50 of 0.87 +/- 0.30 nM, suggesting the expression of the 5-HT1B receptor in these neurons. The presence of 5-HT1B receptors was confirmed by the displacement of [125I]Iodocyanopindolol binding by cyanopindolol with an IC50 of 2.43 +/- 0.81 nM. 5-HT1B receptors are the predominant type of serotonin receptors labeled by [3H]5-HT in cultured DRG neurons, representing approximately 60% of the specific [3H]5-HT binding sites. In addition, 5-HT1D and 5-HT2A receptor binding was also found in these neurons. RT-PCR analysis of RNA isolated from embryonic sensory neurons in culture confirmed the expression of 5-HT1B, 5-HT1D and 5-HT2A receptor mRNA. It also demonstrated the presence of 5-HT1F, 5-HT2C, 5-HT3, 5-HT4, 5-HT5A and 5-HT5B receptor mRNA and the absence of 5-HT1A, 5-HT1E, 5-HT2B, 5-HT6 and 5 HT7 mRNA. The identification of multiple subtypes of serotonin receptors expressed in cultured embryonic sensory neurons suggests that DRG neuronal cultures may be an excellent model to examine the direct effects of serotonin on the activity of these sensory neurons. PMID- 9864302 TI - Investigation of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone for in vivo and iIn vitro murine embryopathy and embryonic ras mutations. AB - The teratological potential of the carcinogen 4-(methylnitrosamino)-1-(3-pyridyl) 1-butanone (NNK) is unknown. In vivo, NNK (100 mg/kg i.p.) was administered to pregnant CD-1 mice during organogenesis, with or without pretreatment with the P450 inducer phenobarbital (60 mg/kg i.p.). With NNK alone, 3 of 374 fetuses had open eye and one had a cleft palate, which were not observed in 160 controls. With phenobarbital plus NNK, two fetuses had a cleft palate, two had exencephaly and one had a kinky tail, although phenobarbital controls showed no anomalies (P <.05). NNK-initiated fetal postpartum lethality was enhanced by phenobarbital pretreatment. There were no fetal skeletal anomalies or alterations in resorptions or fetal body weight in any group. In embryo culture, gestational day 9.5 embryos exposed to 10 microM NNK had decreases in yolk sac diameter, crown rump length and somite development (P <.05), and 100 microM NNK decreased anterior neuropore closure and crown-rump length (P <.05). Embryos exposed to 100 microM NNK were assessed for K-ras codon 12 mutations and none were detected. This is the first evidence for NNK teratogenicity and embryotoxicity, the molecular mechanism of which appears to differ from that for its carcinogenicity. PMID- 9864303 TI - Nicotine evokes cell death in embryonic rat brain during neurulation. AB - Maternal cigarette smoking during pregnancy represents the most prevalent exposure to a suspected neuroteratogen, nicotine. Although animal models have demonstrated brain cell loss and synaptic abnormalities after prenatal nicotine exposure, the multiple effects of nicotine on the maternal-fetal unit make it difficult to prove that nicotine itself is a neuroteratogen. In the current study, whole rat embryo culture was used to study the effects of nicotine at the neural tube stage of development. Beginning on embryonic day 9.5, embryos were exposed to 1, 10 or 100 microM nicotine. After 48 hr, embryos were examined for dysmorphogenesis and were then processed for light microscopic examination of the neuroepithelium. Examination of the forebrain, midbrain and hindbrain regions revealed extensive cytotoxicity, evidenced by cytoplasmic vacuolation, enlargement of intercellular spaces and a sharply increased incidence of pyknotic/apoptotic cells. These alterations were evident in the absence of generalized dysmorphogenesis and were detectable even at the lowest concentration of nicotine. At the highest concentration, abnormalities were present in the majority of cells. Superimposed on cell damage, we found an increase in mitotic figures. Although enhanced mitosis could represent partial compensation for cell loss, the regional selectivity and concentration dependence of the mitogenic effect differed significantly from that of cell death, suggesting separable mechanisms. The present results support the view that nicotine is a neuroteratogen, specifically targeting brain development at concentrations below the threshold for dysmorphogenesis. PMID- 9864304 TI - The centennial before the millennium: the journal of bacteriology as a world voice for microbiological research. PMID- 9864305 TI - Microdermatology: cell surface in the interaction of microbes with the external world. PMID- 9864306 TI - Metabolic alarms and cell division in Escherichia coli. PMID- 9864307 TI - Pkg2, a novel transmembrane protein Ser/Thr kinase of Streptomyces granaticolor. AB - A 4.2-kb SphI-BamHI fragment of chromosomal DNA from Streptomyces granaticolor was cloned and shown to encode a protein with significant sequence similarity to the eukaryotic protein serine/threonine kinases. It consists of 701 amino acids and in the N-terminal part contains all conserved catalytic domains of protein kinases. The C-terminal domain of Pkg2 contains seven tandem repeats of 11 or 12 amino acids with similarity to the tryptophan-docking motif known to stabilize a symmetrical three-dimensional structure called a propeller structure. The pkg2 gene was overexpressed in Escherichia coli, and the gene product (Pkg2) has been found to be autophosphorylated at serine and threonine residues. The N- and C terminal parts of Pkg2 are separated with a hydrophobic stretch of 21 amino acids which translocated a PhoA fusion protein into the periplasm. Thus, Pkg2 is the first transmembrane protein serine/threonine kinase described for streptomycetes. Replacement of the pkg2 gene by the spectinomycin resistance gene resulted in changes in the morphology of aerial hyphae. PMID- 9864308 TI - The Myxococcus xanthus pilQ (sglA) gene encodes a secretin homolog required for type IV pilus biogenesis, social motility, and development. AB - The Myxococcus xanthus sglA1 spontaneous mutation was originally isolated because it allowed dispersed cell growth in liquid yet retained the ability to form fruiting bodies. Consequently, most of today's laboratory strains either contain the sglA1 mutation or were derived from strains that carry it. Subsequent work showed that sglA was a gene for social gliding motility, a process which is mediated by type IV pili. Here sglA is shown to map to the major pil cluster and to encode a 901-amino-acid open reading frame (ORF) that is homologous to the secretin superfamily of proteins. Secretins form a channel in the outer membrane for the transport of macromolecules. The closest homologs found were PilQ proteins from Pseudomonas aeruginosa and Neisseria gonorrhoeae, which are required for type IV pili biogenesis and twitching motility. To signify these molecular and functional similarities, we have changed the name of sglA to pilQ. The hypomorphic pilQ1 (sglA1) allele was sequenced and found to contain two missense mutations at residues 741 (G-->S) and 762 (N-->G). In addition, 19 independent social (S)-motility mutations are shown to map to the pilQ locus. In frame deletions of pilQ and its downstream gene, orfL, were constructed. pilQ is shown to be essential for pilus biogenesis, S-motility, rippling, and fruiting body formation, while orfL is dispensable for these processes. The pilQ1 allele, but not the DeltapilQ allele, was found to render cells hypersensitive to vancomycin, suggesting that PilQ1 alters the permeability properties of the outer membrane. Many differences between pilQ1 and pilQ+ strains have been noted in the literature. We discuss some of these observations and how they may be rationalized in the context of our molecular and functional findings. PMID- 9864309 TI - Roles of chemosensory pathways in transient changes in swimming speed of Rhodobacter sphaeroides induced by changes in photosynthetic electron transport. AB - The response of free-swimming Rhodobacter sphaeroides to increases and decreases in the intensity of light of different wavelengths was analyzed. There was a transient (1 to 2 s) increase in swimming speed in response to an increase in light intensity, and there was a similar transient stop when the light intensity decreased. Measurement of changes in membrane potential and the use of electron transport inhibitors showed that the transient increase in swimming speed, following an increase in light intensity, and the stop following its decrease were the result of changes in photosynthetic electron transport. R. sphaeroides has two operons coding for multiple homologs of the enteric chemosensory genes. Mutants in the first chemosensory operon showed wild-type photoresponses. Mutants with the cheA gene of the second operon (cheAII) deleted, either with or without the first operon present, showed inverted photoresponses, with free-swimming cells stopping on an increase in light intensity and increasing swimming speed on a decrease. These mutants also lacked adaptation. Transposon mutants with mutations in cheAII, which also reduced expression of downstream genes, however, showed no photoresponses. These results show that (i) free-swimming cells respond to both an increase and a decrease in light intensity (tethered cells only show the stopping on a step down in light intensity), (ii) the signal comes from photosynthetic electron transfer, and (iii) the signal is primarily channelled through the second chemosensory pathway. The different responses shown by the cheAII deletion and insertion mutants suggest that CheWII is required for photoresponses, and a third sensory pathway can substitute for CheAII as long as CheWII is present. The inverted response suggests that transducers are involved in photoresponses as well as chemotactic responses. PMID- 9864310 TI - Evidence for a chemiosmotic model of dehalorespiration in Desulfomonile tiedjei DCB-1. AB - Desulfomonile tiedjei DCB-1, a sulfate-reducing bacterium, conserves energy for growth from reductive dehalogenation of 3-chlorobenzoate by an uncharacterized chemiosmotic process. Respiratory electron transport components were examined in D. tiedjei cells grown under conditions for reductive dehalogenation, pyruvate fermentation, and sulfate reduction. Reductive dehalogenation was inhibited by the respiratory quinone inhibitor 2-heptyl-4-hydroxyquinoline N-oxide, suggesting that a respiratory quinoid is a component of the electron transport chain coupled to reductive dehalogenation. Moreover, reductive dehalogenation activity was dependent on 1, 4-naphthoquinone, a possible precursor for a respiratory quinoid. However, no ubiquinone or menaquinone could be extracted from D. tiedjei. Rather, a UV-absorbing quinoid which is different from common respiratory quinones in chemical structure according to mass spectrometric and UV absorption spectroscopic analyses was extracted. ATP sulfurylase, adenosine phosphosulfate reductase, and desulfoviridin sulfite reductase, enzymes involved in sulfate reduction, were constitutively expressed in the cytoplasm of D. tiedjei cells grown under all three metabolic conditions. A periplasmic hydrogenase was detected in cells grown under reductive-dehalogenating and pyruvate-fermenting conditions. A membrane-bound, periplasm-oriented formate dehydrogenase was detected only in cells grown with formate as electron donor, while a cytoplasmic formate dehydrogenase was detected in cells grown under reductive-dehalogenating and pyruvate-fermenting conditions. Results from dehalogenation assays with D. tiedjei whole-cell suspensions and cell extracts suggest that the membrane-bound reductive dehalogenase is cytoplasm oriented. The data clearly demonstrate an enzyme topology in D. tiedjei which produces protons directly in the periplasm, generating a proton motive force by a scalar mechanism. PMID- 9864311 TI - Convergent pathways for utilization of the amino sugars N-acetylglucosamine, N acetylmannosamine, and N-acetylneuraminic acid by Escherichia coli. AB - N-Acetylglucosamine (GlcNAc) and N-acetylneuraminic acid (NANA) are good carbon sources for Escherichia coli K-12, whereas N-acetylmannosamine (ManNAc) is metabolized very slowly. The isolation of regulatory mutations which enhanced utilization of ManNAc allowed us to elucidate the pathway of its degradation. ManNAc is transported by the manXYZ-encoded phosphoenolpyruvate-dependent phosphotransferase system (PTS) transporter producing intracellular ManNAc-6-P. This phosphorylated hexosamine is subsequently converted to GlcNAc-6-P, which is further metabolized by the nagBA-encoded deacetylase and deaminase of the GlcNAc 6-P degradation pathway. Two independent mutations are necessary for good growth on ManNAc. One mutation maps to mlc, and mutations in this gene are known to enhance the expression of manXYZ. The second regulatory mutation was mapped to the nanAT operon, which encodes the NANA transporter and NANA lyase. The combined action of the nanAT gene products converts extracellular NANA to intracellular ManNAc. The second regulatory mutation defines an open reading frame (ORF), called yhcK, as the gene for the repressor of the nan operon (nanR). Mutations in the repressor enhance expression of the nanAT genes and, presumably, three distal, previously unidentified genes, yhcJIH. Expression of just one of these downstream ORFs, yhcJ, is necessary for growth on ManNAc in the presence of an mlc mutation. The yhcJ gene appears to encode a ManNAc-6-P-to-GlcNAc-6-P epimerase (nanE). Another putative gene in the nan operon, yhcI, likely encodes ManNAc kinase (nanK), which should phosphorylate the ManNAc liberated from NANA by the NanA protein. Use of NANA as carbon source by E. coli also requires the nagBA gene products. The existence of a ManNAc kinase and epimerase within the nan operon allows us to propose that the pathways for dissimilation of the three amino sugars GlcNAc, ManNAc, and NANA, all converge at the step of GlcNAc-6-P. PMID- 9864312 TI - Genetic and biochemical characterization of a 2-pyrone-4, 6-dicarboxylic acid hydrolase involved in the protocatechuate 4, 5-cleavage pathway of Sphingomonas paucimobilis SYK-6. AB - Sphingomonas paucimobilis SYK-6 is able to grow on a wide variety of dimeric lignin compounds with guaiacyl moieties, which are converted into protocatechuate by the actions of lignin degradation enzymes in this strain. Protocatechuate is a key metabolite in the SYK-6 degradation of lignin compounds with guaiacyl moieties, and it is thought that it degrades to pyruvate and oxaloacetate via the protocatechuate 4,5-cleavage pathway. In a 10.5-kb EcoRI fragment carrying the protocatechuate 4,5-dioxygenase gene (ligAB) (Y. Noda, S. Nishikawa, K. Shiozuka, H. Kadokura, H. Nakajima, K. Yoda, Y. Katayama, N. Morohoshi, T. Haraguchi, and M. Yamasaki. J. Bacteriol. 172:2704-2709, 1990), we found the ligI gene encoding 2-pyrone-4, 6-dicarboxylic acid (PDC) hydrolase. PDC hydrolase is a member of this pathway and catalyzes the interconversion between PDC and 4-carboxy-2 hydroxymuconic acid (CHM). The ligI gene is thought to be transcribed divergently from ligAB and consists of an 879-bp open reading frame encoding a polypeptide with a molecular mass of 32,737 Da. The ligI gene product (LigI), expressed in Escherichia coli, was purified to near-homogeneity and was estimated to be a monomer (31.6 kDa) by gel filtration chromatography. The isoelectric point was determined to be 4.9. The optimum pH for hydrolysis of PDC is 8.5, the optimum pH for synthesis of PDC is 6.0 to 7.5, and the Km values for PDC and CHM are 74 and 49 microM, respectively. LigI activity was inhibited by the addition of thiol reagents, suggesting that the cysteine residue is a catalytic site. LigI is more resistant to metal ion inhibition than the PDC hydrolases of Pseudomonas ochraceae (K. Maruyama, J. Biochem. 93:557-565, 1983) and Comamonas testosteroni (P. J. Kersten, S. Dagley, J. W. Whittaker, D. M. Arciero, and J. D. Lipscomb, J. Bacteriol. 152:1154-1162, 1982). The insertional inactivation of the ligI gene in S. paucimobilis SYK-6 led to the complete loss of PDC hydrolase activity and to a growth defect on vanillic acid; it did not affect growth on syringic acid. These results indicate that the ligI gene is essential for the growth of SYK-6 on vanillic acid but is not responsible for the growth of SYK-6 on syringic acid. PMID- 9864313 TI - Construction and analysis of a Streptococcus parasanguis recA mutant: homologous recombination is not required for adhesion in an in vitro tooth surface model. AB - PCR was used to amplify an internal region of the recA gene from Streptococcus parasanguis FW213. The PCR fragment was used as a probe to recover the entire streptococcal recA gene from an S. parasanguis genomic library, and the sequence of the gene was determined. The deduced product of the S. parasanguis recA gene showed a high degree of amino acid identity with other prokaryotic RecA proteins. The cloned recA sequence was disrupted in vitro by insertional mutagenesis, and the mutated allele was then introduced into the S. parasanguis chromosome by homologous recombination. Results of Southern hybridizations confirmed the replacement of the wild-type recA gene with the mutated allele. The recA mutant strain was considerably more sensitive to UV light than the parental strain, and this phenotype was consistent with a mutation in recA. The S. parasanguis recA mutant showed no reduction in its ability to adhere in the in vitro tooth surface model, saliva-coated hydroxylapatite (SHA), or in its ability to express the fimbria-associated adhesin Fap1. These results demonstrate that in vitro attachment of S. parasanguis FW213 to SHA and expression of Fap1 are recA independent. PMID- 9864314 TI - Cloning and expression of three new Aazotobacter vinelandii genes closely related to a previously described gene family encoding mannuronan C-5-epimerases. AB - The cloning and expression of a family of five modular-type mannuronan C-5 epimerase genes from Azotobacter vinelandii (algE1 to -5) has previously been reported. The corresponding proteins catalyze the Ca2+-dependent polymer-level epimerization of beta-D-mannuronic acid to alpha-L-guluronic acid (G) in the commercially important polysaccharide alginate. Here we report the identification of three additional structurally similar genes, designated algE6, algE7, and algY. All three genes were sequenced and expressed in Escherichia coli. AlgE6 introduced contiguous stretches of G residues into its substrate (G blocks), while AlgE7 acted as both an epimerase and a lyase. The epimerase activity of AlgE7 leads to formation of alginates with both single G residues and G blocks. AlgY did not display epimerase activity, but a hybrid gene in which the 5' terminal part was exchanged with the corresponding region in algE4 expressed an active epimerase. Southern blot analysis of genomic A. vinelandii DNA, using the 5' part of algE2 as a probe, indicated that all hybridization signals originated from algE1 to -5 or the three new genes reported here. PMID- 9864315 TI - Two distinct mechanisms cause heterogeneity of 16S rRNA. AB - To investigate the frequency of heterogeneity among the multiple 16S rRNA genes within a single microorganism, we determined directly the 120-bp nucleotide sequences containing the hypervariable alpha region of the 16S rRNA gene from 475 Streptomyces strains. Display of the direct sequencing patterns revealed the existence of 136 heterogeneous loci among a total of 33 strains. The heterogeneous loci were detected only in the stem region designated helix 10. All of the substitutions conserved the relevant secondary structure. The 33 strains were divided into two groups: one group, including 22 strains, had less than two heterogeneous bases; the other group, including 11 strains, had five or more heterogeneous bases. The two groups were different in their combinations of heterogeneous bases. The former mainly contained transitional substitutions, and the latter was mainly composed of transversional substitutions, suggesting that at least two mechanisms, possibly misincorporation during DNA replication and horizontal gene transfer, cause rRNA heterogeneity. PMID- 9864316 TI - Multiple small heat shock proteins in rhizobia. AB - Seven genes coding for small heat shock proteins (sHsps) in Bradyrhizobium japonicum have been identified. They are organized in five operons that are coordinately regulated by ROSE, a negatively cis-acting DNA element. The deduced sHsps can be divided into two separate classes: class A, consisting of proteins that show similarity to Escherichia coli IbpA and IbpB, and class B, whose members display significant similarity to other sHsps from prokaryotes and eukaryotes. Two-dimensional gel electrophoresis and Edman sequencing revealed the presence of at least 12 sHsps in B. japonicum, indicating a remarkable abundance of sHsps in this organism. Three additional members of class A and two potentially novel heat shock proteins were identified on the basis of their amino termini. The presence of multiple sHsps was also demonstrated for a variety of Rhizobium and Bradyrhizobium species by immunoblot analysis and two-dimensional gel electrophoresis. An extensive database survey revealed that, in contrast to the rhizobia, other bacteria contain maximally two sHsps whereas many plants have been reported to possess a sHsp superfamily. PMID- 9864317 TI - Characterization of biosynthetic enzymes for ectoine as a compatible solute in a moderately halophilic eubacterium, Halomonas elongata. AB - 1,4,5,6-Tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (ectoine) is an excellent osmoprotectant. The biosynthetic pathway of ectoine from aspartic beta semialdehyde (ASA), in Halomonas elongata, was elucidated by purification and characterization of each enzyme involved. 2,4-Diaminobutyrate (DABA) aminotransferase catalyzed reversively the first step of the pathway, conversion of ASA to DABA by transamination with L-glutamate. This enzyme required pyridoxal 5'-phosphate and potassium ions for its activity and stability. The gel filtration estimated an apparent molecular mass of 260 kDa, whereas molecular mass measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) was 44 kDa. This enzyme exhibited an optimum pH of 8.6 and an optimum temperature of 25 degreesC and had Kms of 9.1 mM for L-glutamate and 4.5 mM for DL-ASA. DABA acetyltransferase catalyzed acetylation of DABA to gamma-N-acetyl alpha,gamma-diaminobutyric acid (ADABA) with acetyl coenzyme A and exhibited an optimum pH of 8.2 and an optimum temperature of 20 degreesC in the presence of 0.4 M NaCl. The molecular mass was 45 kDa by gel filtration. Ectoine synthase catalyzed circularization of ADABA to ectoine and exhibited an optimum pH of 8.5 to 9.0 and an optimum temperature of 15 degreesC in the presence of 0.5 M NaCl. This enzyme had an apparent molecular mass of 19 kDa by SDS-PAGE and a Km of 8.4 mM in the presence of 0. 77 M NaCl. DABA acetyltransferase and ectoine synthase were stabilized in the presence of NaCl (>2 M) and DABA (100 mM) at temperatures below 30 degreesC. PMID- 9864318 TI - Thioredoxin is involved in oxygen-regulated formation of the photosynthetic apparatus of Rhodobacter sphaeroides. AB - Thioredoxin, a redox active protein, has been previously demonstrated to be essential for growth of the anoxygenic photosynthetic bacterium Rhodobacter sphaeroides. In the present study, the involvement of thioredoxin in the formation of the photosynthetic apparatus of R. sphaeroides WS8 was investigated by construction and analysis of a mutant strain disrupted for the chromosomal trxA copy and carrying a plasmid-borne copy of trxA under the control of the hybrid ptrc promoter inducible by IPTG (isopropyl-beta-D-thiogalactopyranoside). This strain was viable in the absence of IPTG but was affected in pigmentation. When shifted from high to low oxygen tension conditions, the trxA mutant showed a reduced bacteriochlorophyll content in comparison to that of the wild type. Although thioredoxin is able to regulate aminolevulinic acid (ALA) synthase (the first enzyme of the tetrapyrrole biosynthetic pathway) activity by a dithiol disulfide exchange, our mutant strain exhibited a level of ALA synthase activity identical to that of the wild type, suggesting that thioredoxin is involved in other steps to regulate the synthesis of the photosynthetic apparatus. Accordingly, we showed that the trxA mutation affects the oxygen-regulated expression of the puf operon encoding the pigment-binding proteins of the light harvesting and reaction center complexes. Upon transition from aerobic to semiaerobic growth conditions, the maximal puf mRNA level was found to be 40 to 50% lower in the mutant strain than in the wild type. The stability of the puf transcripts was identical in both strains grown under low oxygen tension, indicating that the role of thioredoxin in regulating puf expression occurs at the transcriptional level. PMID- 9864319 TI - Identification of an Escherichia coli pepA homolog and its involvement in suppression of the algB phenotype in mucoid Pseudomonas aeruginosa. AB - Strains of Pseudomonas aeruginosa isolated from the respiratory tracts of patients with cystic fibrosis often display a mucoid morphology due to high levels of expression of the exopolysaccharide alginate. The response regulator AlgB is required for full transcription of the alginate biosynthetic operon. Repeated attempts to demonstrate a direct interaction between AlgB and the promoter region of algD, the first gene in the alginate operon, have thus far been unsuccessful. The possibility that AlgB exerts its effect on algD indirectly exists. To identify putative genes under the control of AlgB which affect algD transcription, transposon mutagenesis of nonmucoid algB derivatives of the mucoid strain FRD1 was employed. Of approximately 3,000 transposon mutants screened, 6 were found to display phenotypes which were mucoid relative to the phenotype of the parental algB strain. The phenotypes of these mutants ranged from being only slightly mucoid to being indistinguishable from that of the original FRD1 strain. One of the particularly mucoid transposon mutants was chosen for further study. This strain was found to be disrupted in a previously uncharacterized open reading frame with 56% amino acid identity to PepA of Escherichia coli. PepA is classified as a leucine aminopeptidase, and homologs have been detected in a number of bacterial, plant, and animal species. This novel gene has been designated phpA (P. aeruginosa homolog of pepA). The insertional inactivation of phpA was found to correlate with the mucoid phenotype and an increase in algD transcription in the algB strain. Expression of phpA from an ectopic chromosomal locus compensated for the transposon insertion in the native phpA gene, restoring algD transcription to levels similar to those observed in the parental algB strain. While phpA expression did not appear to be under the control of AlgB at the transcriptional level, this study demonstrates that loss of phpA in an algB genetic background had a positive effect on alginate expression and, more specifically, on transcription of the alginate biosynthetic operon. PMID- 9864320 TI - Chromosome map of Xanthomonas campestris pv. campestris 17 with locations of genes involved in xanthan gum synthesis and yellow pigmentation. AB - No plasmid was detected in Xanthomonas campestris pv. campestris 17, a strain of the causative agent of black rot in cruciferous plants isolated in Taiwan. Its chromosome was cut by PacI, PmeI, and SwaI into five, two, and six fragments, respectively, and a size of 4.8 Mb was estimated by summing the fragment lengths in these digests. Based on the data obtained from partial digestion and Southern hybridization using probes common to pairs of the overlapping fragments or prepared from linking fragments, a circular physical map bearing the PacI, PmeI, and SwaI sites was constructed for the X. campestris pv. campestris 17 chromosome. Locations of eight eps loci involved in exopolysaccharide (xanthan gum) synthesis, two rrn operons each possessing an unique I-CeuI site, one pig cluster required for yellow pigmentation, and nine auxotrophic markers were determined, using mutants isolated by mutagenesis with Tn5(pfm)CmKm. This transposon contains a polylinker with sites for several rare-cutting restriction endonucleases located between the chloramphenicol resistance and kanamycin resistance (Kmr) genes, which upon insertion introduced additional sites into the chromosome. The recA and tdh genes, with known sequences, were mapped by tagging with the polylinker-Kmr segment from Tn5(pfm)CmKm. This is the first map for X. campestris and would be useful for genetic studies of this and related Xanthomonas species. PMID- 9864321 TI - Roles of low-molecular-weight penicillin-binding proteins in Bacillus subtilis spore peptidoglycan synthesis and spore properties. AB - The peptidoglycan cortex of endospores of Bacillus species is required for maintenance of spore dehydration and dormancy, and the structure of the cortex may also allow it to function in attainment of spore core dehydration. A significant difference between spore and growing cell peptidoglycan structure is the low degree of peptide cross-linking in cortical peptidoglycan; regulation of the degree of this cross-linking is exerted by D,D-carboxypeptidases. We report here the construction of mutant B. subtilis strains lacking all combinations of two and three of the four apparent D, D-carboxypeptidases encoded within the genome and the analysis of spore phenotypic properties and peptidoglycan structure for these strains. The data indicate that while the dacA and dacC products have no significant role in spore peptidoglycan formation, the dacB and dacF products both function in regulating the degree of cross-linking of spore peptidoglycan. The spore peptidoglycan of a dacB dacF double mutant was very highly cross-linked, and this structural modification resulted in a failure to achieve normal spore core dehydration and a decrease in spore heat resistance. A model for the specific roles of DacB and DacF in spore peptidoglycan synthesis is proposed. PMID- 9864323 TI - The Azotobacter vinelandii response regulator AlgR is essential for cyst formation. AB - Azotobacter vinelandii produces the exopolysaccharide alginate, which is essential for the encystment process. In Pseudomonas aeruginosa, as well as in A. vinelandii, the sigmaE factor encoded by algU is required for transcription of algD, which encodes a key enzyme of the alginate biosynthetic pathway. The P. aeruginosa response regulator AlgR activates transcription of algD. fimS, located upstream algR, is proposed to encode the AlgR cognate sensor kinase. We have cloned and characterized the A. vinelandii algR gene; the deduced amino acid sequence of the protein encoded by this gene shows 79% identity with its P. aeruginosa homolog. Sequence analysis around the algR gene revealed the absence of a fimS homolog. Inactivation of A. vinelandii algR diminished alginate production by 50%, but did not affect algD transcription, and completely impaired the capacity to form mature cysts. Electron microscopy of the cyst structures formed by the algR mutant revealed that the encystment process is blocked at the step of exine formation. The transcriptional regulation of the A. vinelandii algR gene and the role of AlgR in alginate production differ significantly from those of its P. aeruginosa counterparts. These differences could be due to the fact that in A. vinelandii, alginate plays a role in encystment, a function not found in P. aeruginosa. PMID- 9864322 TI - Molecular and biochemical characterization of the protein template controlling biosynthesis of the lipopeptide lichenysin. AB - Lichenysins are surface-active lipopeptides with antibiotic properties produced nonribosomally by several strains of Bacillus licheniformis. Here, we report the cloning and sequencing of an entire 26.6-kb lichenysin biosynthesis operon from B. licheniformis ATCC 10716. Three large open reading frames coding for peptide synthetases, designated licA, licB (three modules each), and licC (one module), could be detected, followed by a gene, licTE, coding for a thioesterase-like protein. The domain structure of the seven identified modules, which resembles that of the surfactin synthetases SrfA-A to -C, showed two epimerization domains attached to the third and sixth modules. The substrate specificity of the first, fifth, and seventh recombinant adenylation domains of LicA to -C (cloned and expressed in Escherichia coli) was determined to be Gln, Asp, and Ile (with minor Val and Leu substitutions), respectively. Therefore, we suppose that the identified biosynthesis operon is responsible for the production of a lichenysin variant with the primary amino acid sequence L-Gln-L-Leu-D-Leu-L-Val-L-Asp-D-Leu L-Ile, with minor Leu and Val substitutions at the seventh position. PMID- 9864324 TI - 3-nitroadipate, a metabolic intermediate for mineralization of 2, 4-dinitrophenol by a new strain of a Rhodococcus species. AB - The bacterial strain RB1 has been isolated by enrichment cultivation with 2,4 dinitrophenol as the sole nitrogen, carbon, and energy source and characterized, on the basis of 16S rRNA gene sequence comparison, as a Rhodococcus species closely related to Rhodococcus opacus. Rhodococcus sp. strain RB1 degrades 2,4 dinitrophenol, releasing the two nitro groups from the compound as nitrite. The release of nitro groups from 2,4-dinitrophenol occurs in two steps. First, the 2 nitro group is removed as nitrite, with the production of an aliphatic nitro compound identified by 1H nuclear magnetic resonance and mass spectrometry as 3 nitroadipate. Then, this metabolic derivative is further metabolized, releasing its nitro group as nitrite. Full nitrite assimilation upon reduction to ammonia requires that an additional carbon source be supplied to the medium. PMID- 9864325 TI - Sequence diversity of flagellin (fliC) alleles in pathogenic Escherichia coli. AB - To study the molecular evolution of flagellin, the protein subunit specifying flagellar (H) antigens, the fliC genes from 15 pathogenic strains of Escherichia coli were amplified by PCR and sequenced. Comparison of fliC sequences of H6 and H7 strains revealed that alleles have a mosaic structure indicating the occurrence of past horizontal transfer of DNA segments between strains. The close similarity of H7 sequences also indicates the exchange of an entire fliC H7 allele between distant clonal lineages. In addition, the ratio of silent substitutions to amino acid replacements suggests that a short segment in the central region of fliC has been under positive selection in the divergence of H6 and H7 alleles. Phylogenetic analysis demonstrates that the fliC sequences of O157:H7 and O55:H7 serotypes are nearly identical and highly divergent from those of E. coli strains expressing H6 and H2 flagellar antigens. A nonmotile clone of sorbitol-fermenting O157 has rapidly accumulated multiple mutations in fliC, presumably as a result of the silencing of flagellin expression. PMID- 9864326 TI - Kinetics of nirS expression (cytochrome cd1 nitrite reductase) in Pseudomonas stutzeri during the transition from aerobic respiration to denitrification: evidence for a denitrification-specific nitrate- and nitrite-responsive regulatory system. AB - After shifting an oxygen-respiring culture of Pseudomonas stutzeri to nitrate or nitrite respiration, we directly monitored the expression of the nirS gene by mRNA analysis. nirS encodes the 62-kDa subunit of the homodimeric cytochrome cd1 nitrite reductase involved in denitrification. Information was sought about the requirements for gene activation, potential regulators of such activation, and signal transduction pathways triggered by the alternative respiratory substrates. We found that nirS, together with nirT and nirB (which encode tetra- and diheme cytochromes, respectively), is part of a 3.4-kb operon. In addition, we found a 2 kb monocistronic transcript. The half-life of each of these messages was approximately 13 min in denitrifying cells with a doubling time of around 2.5 h. When the culture was subjected to a low oxygen tension, we observed a transient expression of nirS lasting for about 30 min. The continued transcription of the nirS operon required the presence of nitrate or nitrite. This anaerobically manifested N-oxide response was maintained in nitrate sensor (NarX) and response regulator (NarL) knockout strains. Similar mRNA stability and transition kinetics were observed for the norCB operon, encoding the NO reductase complex, and the nosZ gene, encoding nitrous oxide reductase. Our results suggest that a nitrate- and nitrite-responsive regulatory circuit independent of NarXL is necessary for the activation of denitrification genes. PMID- 9864327 TI - Recruitment of ZipA to the septal ring of Escherichia coli is dependent on FtsZ and independent of FtsA. AB - Cell division in prokaryotes is mediated by the septal ring. In Escherichia coli, this organelle consists of several essential division proteins, including FtsZ, FtsA, and ZipA. To gain more insight into how the structure is assembled, we studied the interdependence of FtsZ, FtsA, and ZipA localization using both immunofluorescence and Gfp tagging techniques. To this end, we constructed a set of strains allowing us to determine the cellular location of each of these three proteins in cells from which one of the other two had been specifically depleted. Our results show that ZipA fails to accumulate in a ring shape in the absence of FtsZ. Conversely, depletion of ZipA does not abolish formation of FtsZ rings but leads to a significant reduction in the number of rings per unit of cell mass. In addition, ZipA does not appear to require FtsA for assembly into the septal ring and vice versa. It is suggested that septal ring formation starts by assembly of the FtsZ ring, after which ZipA and FtsA join this structure in a mutually independent fashion through direct interactions with the FtsZ protein. PMID- 9864328 TI - Mutations affecting the ability of the Escherichia coli UmuD' protein to participate in SOS mutagenesis. AB - The products of the SOS-regulated umuDC operon are required for most UV and chemical mutagenesis in Escherichia coli, a process that results from a translesion synthesis mechanism. The UmuD protein is activated for its role in mutagenesis by a RecA-facilitated autodigestion that removes the N-terminal 24 amino acids. A previous genetic screen for nonmutable umuD mutants had resulted in the isolation of a set of missense mutants that produced UmuD proteins that were deficient in RecA-mediated cleavage (J. R. Battista, T. Ohta, T. Nohmi, W. Sun, and G. C. Walker, Proc. Natl. Acad. Sci. USA 87:7190-7194, 1990). To identify elements of the UmuD' protein necessary for its role in translesion synthesis, we began with umuD', a modified form of the umuD gene that directly encodes the UmuD' protein, and obtained missense umuD' mutants deficient in UV and methyl methanesulfonate mutagenesis. The D39G, L40R, and T51I mutations affect residues located at the UmuD'2 homodimer interface and interfere with homodimer formation in vivo. The D75A mutation affects a highly conserved residue located at one end of the central strand in a three-stranded beta-sheet and appears to interfere with UmuD'2 homodimer formation indirectly by affecting the structure of the UmuD' monomer. When expressed from a multicopy plasmid, the L40R umuD' mutant gene exhibited a dominant negative effect on a chromosomal umuD+ gene with respect to UV mutagenesis, suggesting that the mutation has an effect on UmuD' function that goes beyond its impairment of homodimer formation. The G129D mutation affects a highly conserved residue that lies at the end of the long C-terminal beta-strand and results in a mutant UmuD' protein that exhibits a strongly dominant negative effect on UV mutagenesis in a umuD+ strain. The A30V and E35K mutations alter residues in the N-terminal arms of the UmuD'2 homodimer, which are mobile in solution. PMID- 9864329 TI - pSa causes oncogenic suppression of Agrobacterium by inhibiting VirE2 protein export. AB - When coresident with the Ti (tumor-inducing) plasmid, the 21-kDa product of the osa gene of the plasmid pSa can suppress crown gall tumorigenesis incited by Agrobacterium tumefaciens. Neither T-DNA processing nor vir (virulence) gene induction is affected by the presence of osa in the bacterium. We used Arabidopsis thaliana root segments and tobacco leaf discs to demonstrate that Osa inhibits A. tumefaciens from transforming these plants to the stable phenotypes of tumorigenesis, kanamycin resistance, and stable beta-glucuronidase (GUS) expression. When A. tumefaciens contained osa, the lack of expression of transient GUS activity in infected plant tissues, as well as the lack of systemic viral symptoms following agroinfection of Nicotiana benthamiana by tomato mottle virus, suggested that oncogenic suppression by Osa occurs before T-DNA enters the plant nucleus. The extracellular complementation of an A. tumefaciens virE2 mutant (the T-DNA donor strain) by an A. tumefaciens strain lacking T-DNA but containing a wild-type virE2 gene (the VirE2 donor strain) was blocked when osa was present in the VirE2 donor strain, but not when osa was present in the T-DNA donor strain. These data indicate that osa inhibits VirE2 protein, but not T-DNA export from A. tumefaciens. These data further suggest that VirE2 protein and T DNA are separately exported from the bacterium. The successful infection of Datura stramonium plants and leaf discs of transgenic tobacco plants expressing VirE2 protein by an A. tumefaciens virE2 mutant carrying osa confirmed that oncogenic suppression by osa does not occur by blocking T-DNA transfer. Overexpression of virB9, virB10, and virB11 in A. tumefaciens did not overcome oncogenic suppression by osa. The finding that the expression of the osa gene by itself, rather than the formation of a conjugal intermediate with pSa, blocks transformation suggests that the mechanism of oncogenic suppression by osa may differ from that of the IncQ plasmid RSF1010. PMID- 9864330 TI - Protein mobility in the cytoplasm of Escherichia coli. AB - The rate of protein diffusion in bacterial cytoplasm may constrain a variety of cellular functions and limit the rates of many biochemical reactions in vivo. In this paper, we report noninvasive measurements of the apparent diffusion coefficient of green fluorescent protein (GFP) in the cytoplasm of Escherichia coli. These measurements were made in two ways: by photobleaching of GFP fluorescence and by photoactivation of a red-emitting fluorescent state of GFP (M. B. Elowitz, M. G. Surette, P. E. Wolf, J. Stock, and S. Leibler, Curr. Biol. 7:809-812, 1997). The apparent diffusion coefficient, Da, of GFP in E. coli DH5alpha was found to be 7.7 +/- 2.5 microm2/s. A 72-kDa fusion protein composed of GFP and a cytoplasmically localized maltose binding protein domain moves more slowly, with Da of 2.5 +/- 0.6 microm2/s. In addition, GFP mobility can depend strongly on at least two factors: first, Da is reduced to 3.6 +/- 0.7 microm2/s at high levels of GFP expression; second, the addition to GFP of a small tag consisting of six histidine residues reduces Da to 4.0 +/- 2.0 microm2/s. Thus, a single effective cytoplasmic viscosity cannot explain all values of Da reported here. These measurements have implications for the understanding of intracellular biochemical networks. PMID- 9864332 TI - Identification and properties of the genes encoding microcin E492 and its immunity protein. AB - The gene coding for the immunity protein (mceB) and the structural gene of microcin E492 (mceA), a low-molecular-weight channel-forming bacteriocin produced by a strain of Klebsiella pneumoniae, have been characterized. The microcin gene codes for a precursor protein of either 99 or 103 amino acids. Protein sequencing of the N-terminal region of microcin E492 unequivocally identified this gene as the microcin structural gene and indicated that this microcin is synthesized as a precursor protein that is cleaved at either amino acid 15 or 19, at a site resembling the double-glycine motif. The gene encoding the 95-amino-acid immunity protein (mceB) was identified by cloning the DNA segment that encodes only this polypeptide into an expression vector and demonstrating the acquisition of immunity to microcin E492. As expected, the immunity protein was found to be associated with the inner membrane. Analysis of the DNA sequence indicates that these genes belong to the same family as microcin 24, and they do not share structural motifs with any other known channel-forming bacteriocin. The organization of the microcin- and immunity protein-encoding genes suggests that they are coordinately expressed. PMID- 9864331 TI - Evidence that the extracytoplasmic function sigma factor sigmaE is required for normal cell wall structure in Streptomyces coelicolor A3(2). AB - The sigE gene of Streptomyces coelicolor A3(2) encodes an RNA polymerase sigma factor belonging to the extracytoplasmic function (ECF) subfamily. Constructed sigE deletion and disruption mutants were more sensitive than the parent to muramidases such as hen egg white lysozyme and to the CwlA amidase from Bacillus subtilis. This correlated with an altered muropeptide profile, as determined by reverse-phase high-performance liquid chromatography analysis of lytic digests of purified peptidoglycan. The sigE mutants required high levels of magnesium for normal growth and sporulation, overproducing the antibiotic actinorhodin and forming crenellated colonies in its absence. Together, these data suggest that sigE is required for normal cell wall structure. The role of sigmaE was further investigated by analyzing the expression of hrdD, which is partially sigE dependent. The hrdD gene, which encodes the sigmaHrdD subunit of RNA polymerase, is transcribed from two promoters, hrdDp1 and hrdDp2, both similar to promoters recognized by other ECF sigma factors. The activities of hrdDp1 and hrdDp2 were reduced 20- and 3-fold, respectively, in sigE mutants, although only hrdDp1 was recognized by EsigmaE in vitro. Growth on media deficient in magnesium caused the induction of both hrdD promoters in a sigE-dependent manner. PMID- 9864333 TI - Mutations that extend the specificity of the endonuclease activity of lambda terminase. AB - Terminase, an enzyme encoded by the Nu1 and A genes of bacteriophage lambda, is crucial for packaging concatemeric DNA into virions. cosN, a 22-bp segment, is the site on the virus chromosome where terminase introduces staggered nicks to cut the concatemer to generate unit-length virion chromosomes. Although cosN is rotationally symmetric, mutations in cosN have asymmetric effects. The cosN G2C mutation (a G-to-C change at position 2) in the left half of cosN reduces the phage yield 10-fold, whereas the symmetric mutation cosN C11G, in the right half of cosN, does not affect the burst size. The reduction in phage yield caused by cosN G2C is correlated with a defect in cos cleavage. Three suppressors of the cosN G2C mutation, A-E515G, A-N509K, and A-R504C, have been isolated that restore the yield of lambda cosN G2C to the wild-type level. The suppressors are missense mutations that alter amino acids located near an ATPase domain of gpA. lambda A E515G, A-N509K, and A-R504C phages, which are cosN+, also had wild-type burst sizes. In vitro cos cleavage experiments on cosN G2C C11G DNA showed that the rate of cleavage for A-E515G terminase is three- to fourfold higher than for wild type terminase. The A-E515G mutation changes residue 515 of gpA from glutamic acid to glycine. Uncharged polar and hydrophobic residues at position 515 suppressed the growth defect of lambda cosN G2C C11G. In contrast, basic (K, R) and acidic (E, D) residues at position 515 failed to suppress the growth defect of lambda cosN G2C C11G. In a lambda cosN+ background, all amino acids tested at position 515 were functional. These results suggest that A-E515G plays an indirect role in extending the specificity of the endonuclease activity of lambda terminase. PMID- 9864334 TI - Lactobacillus casei 64H contains a phosphoenolpyruvate-dependent phosphotransferase system for uptake of galactose, as confirmed by analysis of ptsH and different gal mutants. AB - Galactose metabolism in Lactobacillus casei 64H was analyzed by genetic and biochemical methods. Mutants with defects in ptsH, galK, or the tagatose 6 phosphate pathway were isolated either by positive selection using 2-deoxyglucose or 2-deoxygalactose or by an enrichment procedure with streptozotocin. ptsH mutations abolish growth on lactose, cellobiose, N-acetylglucosamine, mannose, fructose, mannitol, glucitol, and ribitol, while growth on galactose continues at a reduced rate. Growth on galactose is also reduced, but not abolished, in galK mutants. A mutation in galK in combination with a mutation in the tagatose 6 phosphate pathway results in sensitivity to galactose and lactose, while a galK mutation in combination with a mutation in ptsH completely abolishes galactose metabolism. Transport assays, in vitro phosphorylation assays, and thin-layer chromatography of intermediates of galactose metabolism also indicate the functioning of a permease/Leloir pathway and a phosphoenolpyruvate-dependent phosphotransferase system (PTS)/tagatose 6-phosphate pathway. The galactose-PTS is induced by growth on either galactose or lactose, but the induction kinetics for the two substrates are different. PMID- 9864335 TI - Isolation of a putative Candida albicans transcriptional regulator involved in pleiotropic drug resistance by functional complementation of a pdr1 pdr3 mutation in Saccharomyces cerevisiae. AB - Three Candida albicans genes, designated FCR (for fluconazole resistance), have been isolated by their ability to complement the fluconazole (FCZ) hypersensitivity of a Saccharomyces cerevisiae mutant lacking the transcription factors Pdr1p and Pdr3p. Overexpression of any of the three FCR genes in the pdr1 pdr3 mutant resulted in increased resistance of the cells to FCZ and cycloheximide and in increased expression of PDR5, a gene coding for a drug efflux transporter of the ATP-binding cassette superfamily and whose transcription is under the control of Pdr1p and Pdr3p. Deletion of PDR5 in the pdr1 pdr3 strain completely abrogated the ability of the three FCR genes to confer FCZ resistance, demonstrating that PDR5 is required for FCR-mediated FCZ resistance in S. cerevisiae. The FCR1 gene encodes a putative 517-amino-acid protein with an N-terminal Zn2C6-type zinc finger motif homologous to that found in fungal zinc cluster proteins, including S. cerevisiae Pdr1p and Pdr3p. We have constructed a C. albicans CAI4-derived mutant strain carrying a homozygous deletion of the FCR1 gene and analyzed its ability to grow in the presence of FCZ. We found that the fcr1Delta/fcr1Delta mutant displays hyperresistance to FCZ and other antifungal drugs compared to the parental CAI4 strain. This hyperresistance could be reversed to wild-type levels by reintroduction of a plasmid-borne copy of FCR1 into the fcr1Delta/fcr1Delta mutant. Taken together, our results indicate that the FCR1 gene behaves as a negative regulator of drug resistance in C. albicans and constitute the first evidence that FCZ resistance can result from the inactivation of a regulatory factor such as Fcr1p. PMID- 9864336 TI - A membrane-bound NAD(P)+-reducing hydrogenase provides reduced pyridine nucleotides during citrate fermentation by Klebsiella pneumoniae. AB - During anaerobic growth of Klebsiella pneumoniae on citrate, 9.4 mmol of H2/mol of citrate (4-kPa partial pressure) was formed at the end of growth besides acetate, formate, and CO2. Upon addition of NiCl2 (36 microM) to the growth medium, hydrogen formation increased about 36% to 14.8 mmol/mol of citrate (6 kPa), and the cell yield increased about 15%. Cells that had been harvested and washed under anoxic conditions exhibited an H2-dependent formation of NAD(P)H in vivo. The reduction of internal NAD(P)+ was also achieved by the addition of formate. In crude extracts, the H2:NAD+ oxidoreductase activity was 0.13 micromol min-1 mg-1, and 76% of this activity was found in the washed membrane fraction. The highest specific activities of the membrane fraction were observed in 50 mM potassium phosphate, with 1.6 micromol of NADPH formed min-1 mg-1 at pH 7.0 and 1.7 micromol of NADH formed min-1 mg-1 at pH 9.5. In the presence of the protonophore carbonyl cyanide m-chlorophenylhydrazone and the Na+/H+ antiporter monensin, the H2-dependent reduction of NAD+ by membrane vesicles decreased only slightly (about 16%). The NADP+- or NAD+-reducing hydrogenases were solubilized from the membranes with the detergent lauryldimethylamine-N-oxide or Triton X 100. NAD(P)H formation with H2 as electron donor, therefore, does not depend on an energized state of the membrane. It is proposed that hydrogen which is formed by K. pneumoniae during citrate fermentation is recaptured by a novel membrane bound, oxygen-sensitive H2:NAD(P)+ oxidoreductase that provides reducing equivalents for the synthesis of cell material. PMID- 9864337 TI - Activities of the Porphyromonas gingivalis PrtP proteinase determined by construction of prtP-deficient mutants and expression of the gene in Bacteroides species. AB - PrtP is a major cysteine proteinase of Porphyromonas gingivalis. The gene encoding this proteinase, prtP, was cloned into the Escherichia coli-Bacteroides shuttle vectors pFD288 and pFD340 and was expressed in Bacteroides cells, apparently under the control of its own promoter, when in pFD288, or a Bacteroides promoter present on pFD340. Proteolytically active PrtP was detected by fibrinogen zymography in cells or spent growth medium of several Bacteroides species harboring the recombinant plasmids. The proteinase was recovered from Bacteroides fragilis ATCC 25285(pFD340-prtP) cells by 3-[(3-cholamidopropyl) dimethyl-ammonio]-1-propanesulfonate (CHAPS) extraction and characterized with regard to exopeptidase specificity and sensitivity to proteinase inhibitors. Lys amidolytic activity, but not Arg-amidolytic activity, was detected. PrtP was activated by cysteine and, to a lesser extent, dithiothreitol, and it was stimulated by glycine-containing compounds. It also was inhibited by Nalpha-p tosyl-L-lysine chloromethyl ketone (TLCK) and, to a lesser extent, H-D-Tyr-L-Pro L-arginyl chloromethyl ketone (YPRCK) and was relatively insensitive to EDTA and leupeptin. Neither B. fragilis ATCC 25285(pFD340-prtP) cells nor the CHAPS extract effected hemagglutination of sheep red blood cells or collagen cleavage, but the cells did cleave gelatin. Furthermore, P. gingivalis W12, ATCC 33277, KDP110, and HG66 with knockout mutations in prtP were constructed by allelic replacement. Unlike the parent strains, the mutant strains produced beige colonies on plates containing sheep blood. These strains also were affected in their ability to effect hemagglutination, cleave collagen, and cleave a Lys specific peptide substrate. This report presents the results of the first characterization of the PrtP proteinase clearly in the absence of any influence by other P. gingivalis proteins and describes the properties of P. gingivalis cells defective in the production of PrtP. PMID- 9864338 TI - Function and regulation of glnA in the methanogenic archaeon Methanococcus maripaludis. AB - The glnA gene in the domains Bacteria and Archaea encodes glutamine synthetase, a universally distributed enzyme that functions in ammonia assimilation and glutamine synthesis. We investigated the regulation and function of glnA in the methanogenic archaeon Methanococcus maripaludis. The deduced amino acid sequence of the gene demonstrated its membership in class GSI-alpha of glutamine synthetases. The gene appeared to be expressed as a monocistronic operon. glnA mRNA levels and specific activities of glutamine synthetase were regulated similarly by nitrogen. Three transcription start sites were identified, corresponding to two overlapping nitrogen-regulated promoters and one weaker constitutive promoter. An inverted repeat immediately upstream of the regulated transcription start sites mediated repression under noninducing conditions. Thus, mutations that altered the sequence of the inverted repeat resulted in derepression. The inverted repeat had sequence similarity with a repeat that we previously identified as the nif operator of M. maripaludis, suggesting a common mechanism of nitrogen regulation. Efforts to produce a glnA null mutant failed, suggesting that glnA is an essential gene in M. maripaludis. PMID- 9864339 TI - Repair of oxidized bases in the extremely radiation-resistant bacterium Deinococcus radiodurans. AB - Deinococcus radiodurans is able to resist and survive extreme DNA damage induced by ionizing radiation and many other DNA-damaging agents. It is believed that it possesses highly efficient DNA repair mechanisms. To characterize the repair pathway of oxidized purines in this bacteria, we have purified, from crude extracts, proteins that recognize these oxidized bases. We report here that D. radiodurans possesses two proteins excising the oxidized purines (formamidopyrimidine and 8-oxoguanine) by a DNA glycosylase-a purinic/apyrimidine lyase mechanism. Moreover, one of those proteins is endowed with a thymine glycol DNA glycosylase activity. One of these proteins could be the homolog of the Escherichia coli Fpg enzyme, which confirms the existence of a base excision repair system in this bacteria. PMID- 9864340 TI - Adaptation of Mycobacterium smegmatis to stationary phase. AB - Mycobacterium tuberculosis can persist for many years within host lung tissue without causing clinical disease. Little is known about the state in which the bacilli survive, although it is frequently referred to as dormancy. Some evidence suggests that cells survive in nutrient-deprived stationary phase. Therefore, we are studying stationary-phase survival of Mycobacterium smegmatis as a model for mycobacterial persistence. M. smegmatis cultures could survive 650 days of either carbon, nitrogen, or phosphorus starvation. In carbon-limited medium, cells entered stationary phase before the carbon source (glycerol) had been completely depleted and glycerol uptake from the medium continued during the early stages of stationary phase. These results suggest that the cells are able to sense when the glycerol is approaching limiting concentrations and initiate a shutdown into stationary phase, which involves the uptake of the remaining glycerol from the medium. During early stationary phase, cells underwent reductive cell division and became more resistant to osmotic and acid stress and pool mRNA stabilized. Stationary-phase cells were also more resistant to oxidative stress, but this resistance was induced during late exponential phase in a cell-density-dependent manner. Upon recovery in fresh medium, stationary-phase cultures showed an immediate increase in protein synthesis irrespective of culture age. Colony morphology variants accumulated in stationary-phase cultures. A flat colony variant was seen in 75% of all long-term-stationary-phase cultures and frequently took over the whole population. Cryo scanning electron microscopy showed that the colony organization was different in flat colony strains, flat colonies appearing less well organized than wild-type colonies. Competition experiments with an exponential-phase-adapted wild-type strain showed that the flat strain had a competitive advantage in stationary phase, as well a providing evidence that growth and cell division occur in stationary-phase cultures of M. smegmatis. These results argue against stationary-phase M. smegmatis cultures entering a quiescent state akin to dormancy but support the idea that they are a dynamic population of cells. PMID- 9864341 TI - An endoglucanase, EglA, from the hyperthermophilic archaeon Pyrococcus furiosus hydrolyzes beta-1,4 bonds in mixed-linkage (1-->3),(1-->4)-beta-D-glucans and cellulose. AB - The eglA gene, encoding a thermostable endoglucanase from the hyperthermophilic archaeon Pyrococcus furiosus, was cloned and expressed in Escherichia coli. The nucleotide sequence of the gene predicts a 319-amino-acid protein with a calculated molecular mass of 35.9 kDa. The endoglucanase has a 19-amino-acid signal peptide but not cellulose-binding domain. The P. furiosus endoglucanase has significant amino acid sequence similarities, including the conserved catalytic nucleophile and proton donor, with endoglucanases from glucosyl hydrolase family 12. The purified recombinant enzyme hydrolyzed beta-1,4 but not beta-1,3 glucosidic linkages and had the highest specific activity on cellopentaose (degree of polymerization [DP] = 5) and cellohexaose (DP = 6) oligosaccharides. To a lesser extent, EglA also hydrolyzed shorter cellodextrins (DP < 5) as well as the amorphous portions of polysaccharides which contain only beta-1,4 bonds such as carboxymethyl cellulose, microcrystalline cellulose, Whatman paper, and cotton linter. The highest specific activity toward polysaccharides occurred with mixed-linkage beta-glucans such as barley beta glucan and lichenan. Kinetics studies with cellooliogsaccharides and p nitrophenyl-cellooligosaccharides indicated that the enzyme had three glucose binding subsites (-I, -II, and -III) for the nonreducing end and two glucose binding subsites (+I and +II) for the reducing end from the scissile glycosidic linkage. The enzyme had temperature and pH optima of 100 degreesC and 6.0, respectively; a half-life of 40 h at 95 degreesC; and a denaturing temperature of 112 degreesC as determined by differential scanning calorimetry. The discovery of a thermostable enzyme with this substrate specificity has implications for both the evolution of enzymes involved in polysaccharide hydrolysis and the occurrence of growth substrates in hydrothermal vent environments. PMID- 9864342 TI - Divalent cation block of inward currents and low-affinity K+ uptake in Saccharomyces cerevisiae. AB - We have used the patch clamp technique to characterize whole-cell currents in spheroplasts isolated from a trk1Delta trk2Delta strain of Saccharomyces cerevisiae which lacks high- and moderate-affinity K+ uptake capacity. In solutions in which extracellular divalent cation concentrations were 0.1 mM, cells exhibited a large inward current. This current was not the result of increasing leak between the glass pipette and membrane, as there was no effect on the outward current. The inward current comprised both instantaneous and time dependent components. The magnitude of the inward current increased with increasing extracellular K+ and negative membrane potential but was insensitive to extracellular anions. Replacing extracellular K+ with Rb+, Cs+, or Na+ only slightly modulated the magnitude of the inward current, whereas replacement with Li+ reduced the inward current by approximately 50%, and tetraethylammonium (TEA+) and choline were relatively impermeant. The inward current was blocked by extracellular Ca2+ and Mg2+ with apparent Kis (at -140 mV) of 363 +/- 78 and 96 +/- 14 microM, respectively. Furthermore, decreasing cytosolic K+ increased the magnitude of the inward current independently of the electrochemical driving force for K+ influx, consistent with regulation of the inward current by cytosolic K+. Uptake of 86Rb+ by intact trk1Delta trk2Delta cells was inhibited by extracellular Ca2+ with a Ki within the range observed for the inward current. Furthermore, increasing extracellular Ca2+ from 0.1 to 20 mM significantly inhibited the growth of these cells. These results are consistent with those of the patch clamp experiments in suggesting that low-affinity uptake of alkali cations in yeast is mediated by a transport system sensitive to divalent cations. PMID- 9864343 TI - Purification, properties, and characterization of recombinant Streptomyces sp. strain C5 DoxA, a cytochrome P-450 catalyzing multiple steps in doxorubicin biosynthesis. AB - DoxA is a cytochrome P-450 monooxygenase involved in the late stages of daunorubicin and doxorubicin biosynthesis that has a broad substrate specificity for anthracycline glycone substrates. Recombinant DoxA was purified to homogeneity from Streptomyces lividans transformed with a plasmid containing the Streptomyces sp. strain C5 doxA gene under the control of the strong SnpR activated snpA promoter. The purified enzyme was a monomeric, soluble protein with an apparent Mr of 47,000. Purified DoxA catalyzed the 13-hydroxylation of 13 deoxydaunorubicin, the 13-oxidation of 13-dihydrocarminomycin and 13 dihydrodaunorubicin, and the 14-hydroxylation of daunorubicin. The pH optimum for heme activation was pH 7.5, and the temperature optimum was 30 degreesC. The kcat/Km values for the oxidation of anthracycline substrates by purified DoxA, incubated with appropriate electron-donating components, were as follows: for 13 deoxydaunorubicin, 22,000 M-1 x s-1; for 13-dihydrodaunorubicin, 14,000 M-1 x s 1; for 13-dihydrocarminomycin, 280 M-1 x s-1; and for daunorubicin, 130 M-1 x s 1. Our results indicate that the conversion of daunorubicin to doxorubicin by this enzyme is not a favored reaction and that the main anthracycline flux through the late steps of the daunorubicin biosynthetic pathway catalyzed by DoxA is likely directed through the 4-O-methyl series of anthracyclines. PMID- 9864344 TI - Doxorubicin overproduction in Streptomyces peucetius: cloning and characterization of the dnrU ketoreductase and dnrV genes and the doxA cytochrome P-450 hydroxylase gene. AB - Doxorubicin-overproducing strains of Streptomyces peucetius ATCC 29050 can be obtained through manipulation of the genes in the region of the doxorubicin (DXR) gene cluster that contains dpsH, the dpsG polyketide synthase gene, the putative dnrU ketoreductase gene, dnrV, and the doxA cytochrome P-450 gene. These five genes were characterized by sequence analysis, and the effects of replacing dnrU, dnrV, doxA, or dpsH with mutant alleles and of doxA overexpression on the production of the principal anthracycline metabolites of S. peucetius were studied. The exact roles of dpsH and dnrV could not be established, although dnrV is implicated in the enzymatic reactions catalyzed by DoxA, but dnrU appears to encode a ketoreductase specific for the C-13 carbonyl of daunorubicin (DNR) and DXR or their biosynthetic precursors. The highest DXR titers were obtained in a dnrX dnrU (N. Lomovskaya, Y. Doi-Katayama, S. Filippini, C. Nastro, L. Fonstein, M. Gallo, A. L. Colombo, and C. R. Hutchinson, J. Bacteriol. 180:2379-2386, 1998) double mutant and a dnrX dnrU dnrH (C. Scotti and C. R. Hutchinson, J. Bacteriol. 178:7316-7321, 1996) triple mutant. Overexpression of doxA in a doxA::aphII mutant resulted in the accumulation of DXR precursors instead of in a notable increase in DXR production. In contrast, overexpression of dnrV and doxA jointly in the dnrX dnrU double mutant or the dnrX dnrU dnrH triple mutant increased the DXR titer 36 to 86%. PMID- 9864345 TI - Regulation of the sol locus genes for butanol and acetone formation in Clostridium acetobutylicum ATCC 824 by a putative transcriptional repressor. AB - A gene (orf1, now designated solR) previously identified upstream of the aldehyde/alcohol dehydrogenase gene aad (R. V. Nair, G. N. Bennett, and E. T. Papoutsakis, J. Bacteriol. 176:871-885, 1994) was found to encode a repressor of the sol locus (aad, ctfA, ctfB and adc) genes for butanol and acetone formation in Clostridium acetobutylicum ATCC 824. Primer extension analysis identified a transcriptional start site 35 bp upstream of the solR start codon. Amino acid comparisons of SolR identified a potential helix-turn-helix DNA-binding motif in the C-terminal half towards the center of the protein, suggesting a regulatory role. Overexpression of SolR in strain ATCC 824(pCO1) resulted in a solvent negative phenotype owing to its deleterious effect on the transcription of the sol locus genes. Inactivation of solR in C. acetobutylicum via homologous recombination yielded mutants B and H (ATCC 824 solR::pO1X) which exhibited deregulated solvent production characterized by increased flux towards butanol and acetone formation, earlier induction of aad, lower overall acid production, markedly improved yields of solvents on glucose, a prolonged solvent production phase, and increased biomass accumulation compared to those of the wild-type strain. PMID- 9864347 TI - Peptidoglycan fine structure of the radiotolerant bacterium Deinococcus radiodurans Sark. AB - Peptidoglycan from Deinococcus radiodurans was analyzed by high-performance liquid chromatography and mass spectrometry. The monomeric subunit was: N acetylglucosamine-N-acetylmuramic acid-L-Ala-D-Glu-(gamma)-L-Orn-[(delta)Gly-Gly] D-Ala-D-Ala. Cross-linkage was mediated by (Gly)2 bridges, and glycan strands were terminated in (1-->6)anhydro-muramic acid residues. Structural relations with the phylogenetically close Thermus thermophilus are discussed. PMID- 9864346 TI - (R)-citramalate synthase in methanogenic archaea. AB - The Methanococcus jannaschii gene MJ1392 was cloned, and its protein product was hyperexpressed in Escherichia coli. The resulting protein was purified and shown to catalyze the condensation of pyruvate and acetyl coenzyme A, with the formation of (R)-citramalate. Thus, this gene (cimA) encodes an (R)-citramalate synthase (CimA). This is the first identification of this enzyme, which is likely involved in the biosynthesis of isoleucine. PMID- 9864348 TI - Exopolysaccharide biosynthesis in Lactococcus lactis NIZO B40: functional analysis of the glycosyltransferase genes involved in synthesis of the polysaccharide backbone. AB - We used homologous and heterologous expression of the glycosyltransferase genes of the Lactococcus lactis NIZO B40 eps gene cluster to determine the activity and substrate specificities of the encoded enzymes and established the order of assembly of the trisaccharide backbone of the exopolysaccharide repeating unit. EpsD links glucose-1-phosphate from UDP-glucose to a lipid carrier, EpsE and EpsF link glucose from UDP-glucose to lipid-linked glucose, and EpsG links galactose from UDP-galactose to lipid-linked cellobiose. Furthermore, EpsJ appeared to be involved in EPS biosynthesis as a galactosyl phosphotransferase or an enzyme which releases the backbone oligosaccharide from the lipid carrier. PMID- 9864350 TI - APT1, but not APT2, codes for a functional adenine phosphoribosyltransferase in Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae has two separate genes (APT1 and APT2) that encode two potentially different forms of adenine phosphoribosyltransferase (APRT). However, genetic analysis indicated that only APT1 could code for a complementing activity. Cloning and expression of both the APT1 and APT2 genes in Escherichia coli showed that although discrete proteins (APRT1 and APRT2) were made by these genes, only APRT1 had detectable APRT activity. Northern and Western blot analyses demonstrated that only APT1 was transcribed and translated under normal physiological conditions in yeast. Phylogenetic analysis revealed that APRT1 and APRT2 are evolutionary closely related and that they arise from a gene duplication event. We conclude that APT1 is the functional gene in S. cerevisiae and that APT2 is a pseudogene. PMID- 9864349 TI - Genetic and biochemical analyses of the tec operon suggest a route for evolution of chlorobenzene degradation genes. AB - The TecA broad-spectrum chlorobenzene dioxygenase of Burkholderia sp. strain PS12 catalyzes the first step in the mineralization of 1,2,4, 5-tetrachlorobenzene. The catabolic genes were localized on a small plasmid that belongs to the IncPbeta incompatibility group. PCR analysis of the genetic environment of the tec genes indicated high similarity to the transposon-organized catabolic tcb chlorobenzene degradation genes of Pseudomonas sp. strain P51. Sequence analysis of the regions flanking the tecA genes revealed an upstream open reading frame (ORF) with high similarity to the todF 2-hydroxy-6-oxo-2,4-heptadienoate hydrolase gene of Pseudomonas putida F1 and a discontinuous downstream ORF showing high similarity to the todE catechol 2,3-dioxygenase gene of strain F1. Both homologues in strain P51 exist only as deletion remnants. We suggest that different genetic events thus led to inactivation of the perturbing meta-cleavage enzymes in strains P51 and PS12 during the evolution of efficient chlorobenzene degradation pathways. Biochemical characterization of TodF-like protein TlpF and a genetically refunctionalized TodE-like protein, TlpE, produced in Escherichia coli provided data consistent with the proposed relationships. PMID- 9864351 TI - A novel Bacillus subtilis gene, antE, temporally regulated and convergent to and overlapping dnaE. AB - A Bacillus subtilis promoter, Px, that functions in a convergent manner with the sigA operon promoter P3 has been found in the sigA operon. Promoter Px is turned on at the same time as promoter P3 during early sporulation. The transcript from promoter Px codes for a small protein with partial homology to the OmpR protein from Escherichia coli and also carries an untranslated sequence at its 3' end that is complementary to the 5' end of the P3 transcript, which codes for the ribosome binding site of dnaE. The gene controlled by Px has been called antE. The expression of antE does not require sigmaB, sigmaE, or sigmaH. Px was transcribed in vitro by the sigmaA holoenzyme and is the seventh promoter to be recognized in the sigmaA operon. A possible role for the antE gene during early sporulation is proposed. PMID- 9864353 TI - INCENP centromere and spindle targeting: identification of essential conserved motifs and involvement of heterochromatin protein HP1. AB - The inner centromere protein (INCENP) has a modular organization, with domains required for chromosomal and cytoskeletal functions concentrated near the amino and carboxyl termini, respectively. In this study we have identified an autonomous centromere- and midbody-targeting module in the amino-terminal 68 amino acids of INCENP. Within this module, we have identified two evolutionarily conserved amino acid sequence motifs: a 13-amino acid motif that is required for targeting to centromeres and transfer to the spindle, and an 11-amino acid motif that is required for transfer to the spindle by molecules that have targeted previously to the centromere. To begin to understand the mechanisms of INCENP function in mitosis, we have performed a yeast two-hybrid screen for interacting proteins. These and subsequent in vitro binding experiments identify a physical interaction between INCENP and heterochromatin protein HP1(Hsalpha). Surprisingly, this interaction does not appear to be involved in targeting INCENP to the centromeric heterochromatin, but may instead have a role in its transfer from the chromosomes to the anaphase spindle. PMID- 9864354 TI - Fission yeast bub1 is a mitotic centromere protein essential for the spindle checkpoint and the preservation of correct ploidy through mitosis. AB - The spindle checkpoint ensures proper chromosome segregation by delaying anaphase until all chromosomes are correctly attached to the mitotic spindle. We investigated the role of the fission yeast bub1 gene in spindle checkpoint function and in unperturbed mitoses. We find that bub1(+) is essential for the fission yeast spindle checkpoint response to spindle damage and to defects in centromere function. Activation of the checkpoint results in the recruitment of Bub1 to centromeres and a delay in the completion of mitosis. We show that Bub1 also has a crucial role in normal, unperturbed mitoses. Loss of bub1 function causes chromosomes to lag on the anaphase spindle and an increased frequency of chromosome loss. Such genomic instability is even more dramatic in Deltabub1 diploids, leading to massive chromosome missegregation events and loss of the diploid state, demonstrating that bub1(+ )function is essential to maintain correct ploidy through mitosis. As in larger eukaryotes, Bub1 is recruited to kinetochores during the early stages of mitosis. However, unlike its vertebrate counterpart, a pool of Bub1 remains centromere-associated at metaphase and even until telophase. We discuss the possibility of a role for the Bub1 kinase after the metaphase-anaphase transition. PMID- 9864355 TI - Saccharomyces cerevisiae Ndc1p is a shared component of nuclear pore complexes and spindle pole bodies. AB - We report a novel connection between nuclear pore complexes (NPCs) and spindle pole bodies (SPBs) revealed by our studies of the Saccharomyces cerevisiae NDC1 gene. Although both NPCs and SPBs are embedded in the nuclear envelope (NE) in yeast, their known functions are quite distinct. Previous work demonstrated that NDC1 function is required for proper SPB duplication (Winey, M., M.A. Hoyt, C. Chan, L. Goetsch, D. Botstein, and B. Byers. 1993. J. Cell Biol. 122:743-751). Here, we show that Ndc1p is a membrane protein of the NE that localizes to both NPCs and SPBs. Indirect immunofluorescence microscopy shows that Ndc1p displays punctate, nuclear peripheral localization that colocalizes with a known NPC component, Nup49p. Additionally, distinct spots of Ndc1p localization colocalize with a known SPB component, Spc42p. Immunoelectron microscopy shows that Ndc1p localizes to the regions of NPCs and SPBs that interact with the NE. The NPCs in ndc1-1 mutant cells appear to function normally at the nonpermissive temperature. Finally, we have found that a deletion of POM152, which encodes an abundant but nonessential nucleoporin, suppresses the SPB duplication defect associated with a mutation in the NDC1 gene. We show that Ndc1p is a shared component of NPCs and SPBs and propose a shared function in the assembly of these organelles into the NE. PMID- 9864356 TI - Functional analysis of Tpr: identification of nuclear pore complex association and nuclear localization domains and a role in mRNA export. AB - Tpr is a 270-kD coiled-coil protein localized to intranuclear filaments of the nuclear pore complex (NPC). The mechanism by which Tpr contributes to the structure and function of the nuclear pore is currently unknown. To gain insight into Tpr function, we expressed the full-length protein and several subdomains in mammalian cell lines and examined their effects on nuclear pore function. Through this analysis, we identified an NH2-terminal domain that was sufficient for association with the nucleoplasmic aspect of the NPC. In addition, we unexpectedly found that the acidic COOH terminus was efficiently transported into the nuclear interior, an event that was apparently mediated by a putative nuclear localization sequence. Ectopic expression of the full-length Tpr caused a dramatic accumulation of poly(A)+ RNA within the nucleus. Similar results were observed with domains that localized to the NPC and the nuclear interior. In contrast, expression of these proteins did not appear to affect nuclear import. These data are consistent with a model in which Tpr is tethered to intranuclear filaments of the NPC by its coiled coil domain leaving the acidic COOH terminus free to interact with soluble transport factors and mediate export of macromolecules from the nucleus. PMID- 9864357 TI - Specific binding of the karyopherin Kap121p to a subunit of the nuclear pore complex containing Nup53p, Nup59p, and Nup170p. AB - We have identified a specific karyopherin docking complex within the yeast nuclear pore complex (NPC) that contains two novel, structurally related nucleoporins, Nup53p and Nup59p, and the NPC core protein Nup170p. This complex was affinity purified from cells expressing a functional Nup53p-protein A chimera. The localization of Nup53p, Nup59p, and Nup170p within the NPC by immunoelectron microscopy suggests that the Nup53p-containing complex is positioned on both the cytoplasmic and nucleoplasmic faces of the NPC core. In association with the isolated complex, we have also identified the nuclear transport factor Kap121p (Pse1p). Using in vitro binding assays, we showed that each of the nucleoporins interacts with one another. However, the association of Kap121p with the complex is mediated by its interaction with Nup53p. Moreover, Kap121p is the only beta-type karyopherin that binds Nup53p suggesting that Nup53p acts as a specific Kap121p docking site. Kap121p can be released from Nup53p by the GTP bound form of the small GTPase Ran. The physiological relevance of the interaction between Nup53p and Kap121p was further underscored by the observation that NUP53 mutations alter the subcellular distribution of Kap121p and the Kap121p- mediated import of a ribosomal L25 reporter protein. Interestingly, Nup53p is specifically phosphorylated during mitosis. This phenomenon is correlated with a transient decrease in perinuclear-associated Kap121p. PMID- 9864358 TI - Homotypic fusion of immature secretory granules during maturation in a cell-free assay. AB - The biogenesis of secretory granules embodies several morphological and biochemical changes. In particular, in neuroendocrine cells maturation of secretory granules is characterized by an increase in size which has been proposed to reflect homotypic fusion of immature secretory granules (ISGs). Here we describe an assay that provides the first biochemical evidence for such a fusion event and allows us to analyze its regulation. The assay reconstitutes homotypic fusion between one population of ISGs containing a [35S]sulfate-labeled substrate, secretogranin II (SgII), and a second population containing the prohormone convertase PC2. Both substrate and enzyme are targeted exclusively to ISGs. Fusion is measured by quantification of a cleavage product of SgII produced by PC2. With this assay we show that fusion only occurs between ISGs and not between ISGs and MSGs, is temperature dependent, and requires ATP and GTP and cytosolic proteins. NSF (N-ethylmaleimide-sensitive fusion protein) is amongst the cytosolic proteins required, whereas we could not detect a requirement for p97. The ability to reconstitute ISG fusion in a cell-free assay is an important advance towards the identification of molecules involved in the maturation of secretory granules and will increase our understanding of this process. PMID- 9864360 TI - Pex18p and Pex21p, a novel pair of related peroxins essential for peroxisomal targeting by the PTS2 pathway. AB - We have identified ScPex18p and ScPex21p, two novel S. cerevisiae peroxins required for protein targeting via the PTS2 branch of peroxisomal biogenesis. Targeting by this pathway is known to involve the interaction of oligopeptide PTS2 signals with Pex7p, the PTS2 receptor. Pex7p function is conserved between yeasts and humans, with defects in the human protein causing rhizomelic chondrodysplasia punctata (RCDP), a severe, lethal peroxisome biogenesis disorder characterized by aberrant targeting of several PTS2 peroxisomal proteins, but uncertainty remains about the subcellular localization of this receptor. Previously, we have reported that ScPex7p resides predominantly in the peroxisomal matrix, suggesting that it may function as a highly unusual intraorganellar import receptor, and the data presented in this paper identify Pex18p and Pex21p as key components in the targeting of Pex7p to peroxisomes. They each interact specifically with Pex7p both in two-hybrid analyses and in vitro. In cells lacking both Pex18p and Pex21p, Pex7p remains cytosolic and PTS2 targeting is completely abolished. Pex18p and Pex21p are weakly homologous to each other and display partial functional redundancy, indicating that they constitute a two-member peroxin family specifically required for Pex7p and PTS2 targeting. PMID- 9864359 TI - Biochemical and functional studies of cortical vesicle fusion: the SNARE complex and Ca2+ sensitivity. AB - Cortical vesicles (CV) possess components critical to the mechanism of exocytosis. The homotypic fusion of CV centrifuged or settled into contact has a sigmoidal Ca2+ activity curve comparable to exocytosis (CV-PM fusion). Here we show that Sr2+ and Ba2+ also trigger CV-CV fusion, and agents affecting different steps of exocytotic fusion block Ca2+, Sr2+, and Ba2+-triggered CV-CV fusion. The maximal number of active fusion complexes per vesicle, Max, was quantified by NEM inhibition of fusion, showing that CV-CV fusion satisfies many criteria of a mathematical analysis developed for exocytosis. Both Max and the Ca2+ sensitivity of fusion complex activation were comparable to that determined for CV-PM fusion. Using Ca2+-induced SNARE complex disruption, we have analyzed the relationship between membrane fusion (CV-CV and CV-PM) and the SNARE complex. Fusion and complex disruption have different sensitivities to Ca2+, Sr2+, and Ba2+, the complex remains Ca2+- sensitive on fusion-incompetent CV, and disruption does not correlate with the quantified activation of fusion complexes. Under conditions which disrupt the SNARE complex, CV on the PM remain docked and fusion competent, and isolated CV still dock and fuse, but with a markedly reduced Ca2+ sensitivity. Thus, in this system, neither the formation, presence, nor disruption of the SNARE complex is essential to the Ca2+-triggered fusion of exocytotic membranes. Therefore the SNARE complex alone cannot be the universal minimal fusion machine for intracellular fusion. We suggest that this complex modulates the Ca2+ sensitivity of fusion. PMID- 9864361 TI - Redundant and distinct functions for dynamin-1 and dynamin-2 isoforms. AB - A role for dynamin in clathrin-mediated endocytosis is now well established. However, mammals express three closely related, tissue-specific dynamin isoforms, each with multiple splice variants. Thus, an important question is whether these isoforms and splice variants function in vesicle formation from distinct intracellular organelles. There are conflicting data as to a role for dynamin-2 in vesicle budding from the TGN. To resolve this issue, we compared the effects of overexpression of dominant-negative mutants of dynamin-1 (the neuronal isoform) and dynamin-2 (the ubiquitously expressed isoform) on endocytic and biosynthetic membrane trafficking in HeLa cells and polarized MDCK cells. Both dyn1(K44A) and dyn2(K44A) were potent inhibitors of receptor-mediated endocytosis; however neither mutant directly affected other membrane trafficking events, including transport mediated by four distinct classes of vesicles budding from the TGN. Dyn2(K44A) more potently inhibited receptor-mediated endocytosis than dyn1(K44A) in HeLa cells and at the basolateral surface of MDCK cells. In contrast, dyn1(K44A) more potently inhibited endocytosis at the apical surface of MDCK cells. The two dynamin isoforms have redundant functions in endocytic vesicle formation, but can be targeted to and function differentially at subdomains of the plasma membrane. PMID- 9864362 TI - Aggresomes: a cellular response to misfolded proteins. AB - Intracellular deposition of misfolded protein aggregates into ubiquitin-rich cytoplasmic inclusions is linked to the pathogenesis of many diseases. Why these aggregates form despite the existence of cellular machinery to recognize and degrade misfolded protein and how they are delivered to cytoplasmic inclusions are not known. We have investigated the intracellular fate of cystic fibrosis transmembrane conductance regulator (CFTR), an inefficiently folded integral membrane protein which is degraded by the cytoplasmic ubiquitin-proteasome pathway. Overexpression or inhibition of proteasome activity in transfected human embryonic kidney or Chinese hamster ovary cells led to the accumulation of stable, high molecular weight, detergent-insoluble, multiubiquitinated forms of CFTR. Using immunofluorescence and transmission electron microscopy with immunogold labeling, we demonstrate that undegraded CFTR molecules accumulate at a distinct pericentriolar structure which we have termed the aggresome. Aggresome formation is accompanied by redistribution of the intermediate filament protein vimentin to form a cage surrounding a pericentriolar core of aggregated, ubiquitinated protein. Disruption of microtubules blocks the formation of aggresomes. Similarly, inhibition of proteasome function also prevented the degradation of unassembled presenilin-1 molecules leading to their aggregation and deposition in aggresomes. These data lead us to propose that aggresome formation is a general response of cells which occurs when the capacity of the proteasome is exceeded by the production of aggregation-prone misfolded proteins. PMID- 9864363 TI - Visualization of melanosome dynamics within wild-type and dilute melanocytes suggests a paradigm for myosin V function In vivo. AB - Unlike wild-type mouse melanocytes, where melanosomes are concentrated in dendrites and dendritic tips, melanosomes in dilute (myosin Va-) melanocytes are concentrated in the cell center. Here we sought to define the role that myosin Va plays in melanosome transport and distribution. Actin filaments that comprise a cortical shell running the length of the dendrite were found to exhibit a random orientation, suggesting that myosin Va could drive the outward spreading of melanosomes by catalyzing random walks. In contrast to this mechanism, time lapse video microscopy revealed that melanosomes undergo rapid ( approximately 1.5 microm/s) microtubule-dependent movements to the periphery and back again. This bidirectional traffic occurs in both wild-type and dilute melanocytes, but it is more obvious in dilute melanocytes because the only melanosomes in their periphery are those undergoing this movement. While providing an efficient means to transport melanosomes to the periphery, this component does not by itself result in their net accumulation there. These observations, together with previous studies showing extensive colocalization of myosin Va and melanosomes in the actin-rich periphery, suggest a mechanism in which a myosin Va-dependent interaction of melanosomes with F-actin in the periphery prevents these organelles from returning on microtubules to the cell center, causing their distal accumulation. This "capture" model is supported by the demonstration that (a) expression of the myosin Va tail domain within wild-type cells creates a dilute-like phenotype via a process involving initial colocalization of tail domains with melanosomes in the periphery, followed by an approximately 120-min, microtubule-based redistribution of melanosomes to the cell center; (b) microtubule-dependent melanosome movement appears to be damped by myosin Va; (c) intermittent, microtubule-independent, approximately 0.14 microm/s melanosome movements are seen only in wild-type melanocytes; and (d) these movements do not drive obvious spreading of melanosomes over 90 min. We conclude that long-range, bidirectional, microtubule-dependent melanosome movements, coupled with actomyosin Va-dependent capture of melanosomes in the periphery, is the predominant mechanism responsible for the centrifugal transport and peripheral accumulation of melanosomes in mouse melanocytes. This mechanism represents an alternative to straightforward transport models when interpreting other myosin V mutant phenotypes. PMID- 9864365 TI - Tropomyosin-containing actin cables direct the Myo2p-dependent polarized delivery of secretory vesicles in budding yeast. AB - The actin cytoskeleton in budding yeast consists of cortical patches and cables, both of which polarize toward regions of cell growth. Tropomyosin localizes specifically to actin cables and not cortical patches. Upon shifting cells with conditionally defective tropomyosin to restrictive temperatures, actin cables disappear within 1 min and both the unconventional class V myosin Myo2p and the secretory vesicle-associated Rab GTPase Sec4p depolarize rapidly. Bud growth ceases and the mother cell grows isotropically. When returned to permissive temperatures, tropomyosin-containing cables reform within 1 min in polarized arrays. Cable reassembly permits rapid enrichment of Myo2p at the focus of nascent cables as well as the Myo2p- dependent recruitment of Sec4p and the exocyst protein Sec8p, and the initiation of bud emergence. With the loss of actin cables, cortical patches slowly assume an isotropic distribution within the cell and will repolarize only after restoration of cables. Therefore, actin cables respond to polarity cues independently of the overall distribution of cortical patches and are able to directly target the Myo2p-dependent delivery of secretory vesicles and polarization of growth. PMID- 9864364 TI - Visualization and molecular analysis of actin assembly in living cells. AB - Actin filament assembly is critical for eukaryotic cell motility. Arp2/3 complex and capping protein (CP) regulate actin assembly in vitro. To understand how these proteins regulate the dynamics of actin filament assembly in a motile cell, we visualized their distribution in living fibroblasts using green flourescent protein (GFP) tagging. Both proteins were concentrated in motile regions at the cell periphery and at dynamic spots within the lamella. Actin assembly was required for the motility and dynamics of spots and for motility at the cell periphery. In permeabilized cells, rhodamine-actin assembled at the cell periphery and at spots, indicating that actin filament barbed ends were present at these locations. Inhibition of the Rho family GTPase rac1, and to a lesser extent cdc42 and RhoA, blocked motility at the cell periphery and the formation of spots. Increased expression of phosphatidylinositol 5-kinase promoted the movement of spots. Increased expression of LIM-kinase-1, which likely inactivates cofilin, decreased the frequency of moving spots and led to the formation of aggregates of GFP-CP. We conclude that spots, which appear as small projections on the surface by whole mount electron microscopy, represent sites of actin assembly where local and transient changes in the cortical actin cytoskeleton take place. PMID- 9864366 TI - Dual function of Cyk2, a cdc15/PSTPIP family protein, in regulating actomyosin ring dynamics and septin distribution. AB - We previously showed that the budding yeast Saccharomyces cerevisiae assembles an actomyosin-based ring that undergoes a contraction-like size change during cytokinesis. To learn more about the biochemical composition and activity of this ring, we have characterized the in vivo distribution and function of Cyk2p, a budding yeast protein that exhibits significant sequence similarity to the cdc15/PSTPIP family of cleavage furrow proteins. Video microscopy of cells expressing green fluorescent protein (GFP)-tagged Cyk2p revealed that Cyk2p forms a double ring that coincides with the septins through most of the cell cycle. During cytokinesis, however, the Cyk2 double ring merges with the actomyosin ring and exhibits a contraction-like size change that is dependent on Myo1p. The septin double ring, in contrast, does not undergo the contraction-like size change but the separation between the two rings increases during cytokinesis. These observations suggest that the septin-containing ring is dynamically distinct from the actomyosin ring and that Cyk2p transits between the two types of structures. Gene disruption of CYK2 does not affect the assembly of the actomyosin ring but results in rapid disassembly of the ring during the contraction phase, leading to incomplete cytokinesis, suggesting that Cyk2p has an important function in modulating the stability of the actomyosin ring during contraction. Overexpression of Cyk2p also blocks cytokinesis, most likely due to a loss of the septins from the bud neck, indicating that Cyk2p may also play a role in regulating the localization of the septins. PMID- 9864367 TI - Nonuniform microtubular polarity established by CHO1/MKLP1 motor protein is necessary for process formation of podocytes. AB - Podocytes are unique cells that are decisively involved in glomerular filtration. They are equipped with a complex process system consisting of major processes and foot processes whose function is insufficiently understood (Mundel, P., and W. Kriz. 1995. Anat. Embryol. 192:385-397). The major processes of podocytes contain a microtubular cytoskeleton. Taking advantage of a recently established cell culture system for podocytes with preserved ability to form processes (Mundel, P., J. Reiser, A. Zuniga Mejia Borja, H. Pavenstadt, G.R. Davidson, W. Kriz, and R. Zeller. 1997b. Exp. Cell Res. 36:248-258), we studied the functional significance of the microtubular system in major processes. The following data were obtained: (a) Microtubules (MTs) in podocytes show a nonuniform polarity as revealed by hook-decoration. (b) CHO1/ MKLP1, a kinesin-like motor protein, is associated with MTs in podocytes. (c) Treatment of differentiating podocytes with CHO1/MKLP1 antisense oligonucleotides abolished the formation of processes and the nonuniform polarity of MTs. (d) During the recovery from taxol treatment, taxol-stabilized (nocodazole- resistant) MT fragments were distributed in the cell periphery along newly assembled nocodazole-sensitive MTs. A similar distribution pattern of CHO1/MKLP1 was found under these circumstances, indicating its association with MTs. (e) In the recovery phase after complete depolymerization, MTs reassembled exclusively at centrosomes. Taken together, these findings lead to the conclusion that the nonuniform MT polarity in podocytes established by CHO1/MKLP1 is necessary for process formation. PMID- 9864368 TI - A gamete-specific, sex-limited homeodomain protein in Chlamydomonas. AB - During fertilization in Chlamydomonas, gametes of opposite mating types interact with each other through sex-specific adhesion molecules on their flagellar surfaces. Flagellar adhesion brings the cell bodies of the gametes into close contact and initiates a signal transduction pathway in preparation for cell-cell fusion. We have identified a cDNA, gsp1, whose transcript levels are upregulated during flagellar adhesion. The GSP1 polypeptide is a novel, gamete-specific homeodomain protein, the first to be identified in an alga. Its homeodomain shows significant identity with several higher plant homeodomain proteins. Although encoded by a single copy gene present in cells of both mating types, immunoblot analysis showed that GSP1 was expressed in mating type (mt)+ gametes, but was not detectable in mt- gametes or in vegetative cells of either mating type. Moreover, GSP1 appeared late during gametogenesis, suggesting that it may function during adhesion with mt- gametes or after zygote formation. GSP1 is expressed in imp11, mt- mutant gametes, which have a lesion in the mid gene involved in sex determination and exhibit many phenotypic characteristics of mt+ gametes. Thus, gsp1 is negatively regulated by mid and is the first molecule to be identified in Chlamydomonas that shows sex-limited expression. PMID- 9864369 TI - ARF1 mediates paxillin recruitment to focal adhesions and potentiates Rho stimulated stress fiber formation in intact and permeabilized Swiss 3T3 fibroblasts. AB - Focal adhesion assembly and actin stress fiber formation were studied in serum starved Swiss 3T3 fibroblasts permeabilized with streptolysin-O. Permeabilization in the presence of GTPgammaS stimulated rho-dependent formation of stress fibers, and the redistribution of vinculin and paxillin from a perinuclear location to focal adhesions. Addition of GTPgammaS at 8 min after permeabilization still induced paxillin recruitment to focal adhesion-like structures at the ends of stress fibers, but vinculin remained in the perinuclear region, indicating that the distributions of these two proteins are regulated by different mechanisms. Paxillin recruitment was largely rho-independent, but could be evoked using constitutively active Q71L ADP-ribosylation factor (ARF1), and blocked by NH2 terminally truncated Delta17ARF1. Moreover, leakage of endogenous ARF from cells was coincident with loss of GTPgammaS- induced redistribution of paxillin to focal adhesions, and the response was recovered by addition of ARF1. The ability of ARF1 to regulate paxillin recruitment to focal adhesions was confirmed by microinjection of Q71LARF1 and Delta17ARF1 into intact cells. Interestingly, these experiments showed that V14RhoA- induced assembly of actin stress fibers was potentiated by Q71LARF1. We conclude that rho and ARF1 activate complimentary pathways that together lead to the formation of paxillin-rich focal adhesions at the ends of prominent actin stress fibers. PMID- 9864370 TI - Regulation of the cell cycle by focal adhesion kinase. AB - In this report, we have analyzed the potential role and mechanisms of integrin signaling through FAK in cell cycle regulation by using tetracycline-regulated expression of exogenous FAK and mutants. We have found that overexpression of wild-type FAK accelerated G1 to S phase transition. Conversely, overexpression of a dominant-negative FAK mutant DeltaC14 inhibited cell cycle progression at G1 phase and this inhibition required the Y397 in DeltaC14. Biochemical analyses indicated that FAK mutant DeltaC14 was mislocalized and functioned as a dominant negative mutant by competing with endogenous FAK in focal contacts for binding signaling molecules such as Src and Fyn, resulting in a decreases of Erk activation in cell adhesion. Consistent with this, we also observed inhibition of BrdU incorporation and Erk activation by FAK Y397F mutant and FRNK, but not FRNKDeltaC14, in transient transfection assays using primary human foreskin fibroblasts. Finally, we also found that DeltaC14 blocked cyclin D1 upregulation and induced p21 expression, while wild-type FAK increased cyclin D1 expression and decreased p21 expression. Taken together, these results have identified FAK and its associated signaling pathways as a mediator of the cell cycle regulation by integrins. PMID- 9864371 TI - Desmoplakin is required early in development for assembly of desmosomes and cytoskeletal linkage. AB - Desmosomes first assemble in the E3.5 mouse trophectoderm, concomitant with establishment of epithelial polarity and appearance of a blastocoel cavity. Throughout development, they increase in size and number and are especially abundant in epidermis and heart muscle. Desmosomes mediate cell-cell adhesion through desmosomal cadherins, which differ from classical cadherins in their attachments to intermediate filaments (IFs), rather than actin filaments. Of the proteins implicated in making this IF connection, only desmoplakin (DP) is both exclusive to and ubiquitous among desmosomes. To explore its function and importance to tissue integrity, we ablated the desmoplakin gene. Homozygous -/- mutant embryos proceeded through implantation, but did not survive beyond E6.5. Surprisingly, analysis of these embryos revealed a critical role for desmoplakin not only in anchoring IFs to desmosomes, but also in desmosome assembly and/or stabilization. This finding not only unveiled a new function for desmoplakin, but also provided the first opportunity to explore desmosome function during embryogenesis. While a blastocoel cavity formed and epithelial cell polarity was at least partially established in the DP (-/-) embryos, the paucity of desmosomal cell-cell junctions severely affected the modeling of tissue architecture and shaping of the early embryo. PMID- 9864372 TI - Mutation of a major keratin phosphorylation site predisposes to hepatotoxic injury in transgenic mice. AB - Simple epithelia express keratins 8 (K8) and 18 (K18) as their major intermediate filament (IF) proteins. One important physiologic function of K8/18 is to protect hepatocytes from drug-induced liver injury. Although the mechanism of this protection is unknown, marked K8/18 hyperphosphorylation occurs in association with a variety of cell stresses and during mitosis. This increase in keratin phosphorylation involves multiple sites including human K18 serine-(ser)52, which is a major K18 phosphorylation site. We studied the significance of keratin hyperphosphorylation and focused on K18 ser52 by generating transgenic mice that overexpress a human genomic K18 ser52--> ala mutant (S52A) and compared them with mice that overexpress, at similar levels, wild-type (WT) human K18. Abrogation of K18 ser52 phosphorylation did not affect filament organization after partial hepatectomy nor the ability of mouse livers to regenerate. However, exposure of S52A-expressing mice to the hepatotoxins, griseofulvin or microcystin, which are associated with K18 ser52 and other keratin phosphorylation changes, resulted in more dramatic hepatotoxicity as compared with WT K18-expressing mice. Our results demonstrate that K18 ser52 phosphorylation plays a physiologic role in protecting hepatocytes from stress-induced liver injury. Since hepatotoxins are associated with increased keratin phosphorylation at multiple sites, it is likely that unique sites aside from K18 ser52, and phosphorylation sites on other IF proteins, also participate in protection from cell stress. PMID- 9864373 TI - Molecular organization of sarcoglycan complex in mouse myotubes in culture. AB - The sarcoglycans are a complex of four transmembrane proteins (alpha, beta, gamma, and delta) which are primarily expressed in skeletal muscle and are closely associated with dystrophin and the dystroglycans in the muscle membrane. Mutations in the sarcoglycans are responsible for four autosomal recessive forms of muscular dystrophy. The function and the organization of the sarcoglycan complex are unknown. We have used coimmunoprecipitation and in vivo cross-linking techniques to analyze the sarcoglycan complex in cultured mouse myotubes. We demonstrate that the interaction between beta- and delta-sarcoglycan is resistant to high concentrations of SDS and alpha-sarcoglycan is less tightly associated with other members of the complex. Cross-linking experiments show that beta-, gamma-, and delta-sarcoglycan are in close proximity to one another and that delta-sarcoglycan can be cross-linked to the dystroglycan complex. In addition, three of the sarcoglycans (beta, gamma, and delta) are shown to form intramolecular disulfide bonds. These studies further our knowledge of the structure of the sarcoglycan complex. Our proposed model of their interactions helps to explain some of the emerging data on the consequences of mutations in the individual sarcoglycans, their effect on the complex, and potentially the clinical course of muscular dystrophies. PMID- 9864375 TI - Autocrine tumor necrosis factor (TNF) and lymphotoxin (LT) alpha differentially modulate cellular sensitivity to TNF/LT-alpha cytotoxicity in L929 cells. AB - Tumor necrosis factor (TNF) and lymphotoxin (LT) alpha are structurally and functionally related cytokines. We expressed the TNF and LT-alpha genes in murine fibrosarcoma L929r2 cells, which can be sensitized to TNF/LT-alpha-dependent necrosis by inhibitors of transcription or translation. Autocrine production of murine TNF in L929r2 cells completely downmodulated the expression of the 55- and 75-kD TNF receptors, resulting in resistance to TNF/LT-alpha cytotoxicity. Partial downmodulation of the 55-kD receptor was observed in human TNF-producing L929r2 cells. In contrast, an unaltered TNF receptor expression was found on LT alpha L929r2 transfectants. Hence, although similar cytotoxic effects are induced by extracellularly administered TNF and LT-alpha, endogenous expression of these cytokines fundamentally differs in the way they modulate TNF receptor expression. Unlike LT-alpha, secreted by the classical pathway, TNF is first formed as a membrane-bound protein, which is responsible for receptor downmodulation. To explore whether the different pathways for secretion of TNF and LT-alpha explain this difference, we examined the effect of membrane-bound LT-alpha expression. This was obtained by exchange of the classical signal sequence of LT-alpha for the membrane anchor of chicken hepatic lectin. Membrane retention of LT-alpha resulted indeed in receptor downmodulation and TNF/LT-alpha resistance. We conclude that membrane retention of newly synthesized TNF or LT-alpha is absolutely required for receptor downmodulation and TNF/LT-alpha resistance. PMID- 9864374 TI - Functional characteristics of ES cell-derived cardiac precursor cells identified by tissue-specific expression of the green fluorescent protein. AB - In contrast to terminally differentiated cardiomyocytes, relatively little is known about the characteristics of mammalian cardiac cells before the initiation of spontaneous contractions (precursor cells). Functional studies on these cells have so far been impossible because murine embryos of the corresponding stage are very small, and cardiac precursor cells cannot be identified because of the lack of cross striation and spontaneous contractions. In the present study, we have used the murine embryonic stem (ES, D3 cell line) cell system for the in vitro differentiation of cardiomyocytes. To identify the cardiac precursor cells, we have generated stably transfected ES cells with a vector containing the gene of the green fluorescent protein (GFP) under control of the cardiac alpha-actin promoter. First, fluorescent areas in ES cell-derived cell aggregates (embryoid bodies [EBs]) were detected 2 d before the initiation of contractions. Since Ca2+ homeostasis plays a key role in cardiac function, we investigated how Ca2+ channels and Ca2+ release sites were built up in these GFP-labeled cardiac precursor cells and early stage cardiomyocytes. Patch clamp and Ca2+ imaging experiments proved the functional expression of the L-type Ca2+ current (ICa) starting from day 7 of EB development. On day 7, using 10 mM Ca2+ as charge carrier, ICa was expressed at very low densities 4 pA/pF. The biophysical and pharmacological properties of ICa proved similar to terminally differentiated cardiomyocytes. In cardiac precursor cells, ICa was found to be already under control of cAMP-dependent phosphorylation since intracellular infusion of the catalytic subunit of protein kinase A resulted in a 1.7-fold stimulation. The adenylyl cyclase activator forskolin was without effect. IP3-sensitive intracellular Ca2+ stores and Ca2+-ATPases are present during all stages of differentiation in both GFP-positive and GFP-negative cells. Functional ryanodine sensitive Ca2+ stores, detected by caffeine-induced Ca2+ release, appeared in most GFP-positive cells 1-2 d after ICa. Coexpression of both ICa and ryanodine sensitive Ca2+ stores at day 10 of development coincided with the beginning of spontaneous contractions in most EBs. Thus, the functional expression of voltage dependent L-type Ca2+ channel (VDCC) is a hallmark of early cardiomyogenesis, whereas IP3 receptors and sarcoplasmic Ca2+-ATPases are expressed before the initiation of cardiomyogenesis. Interestingly, the functional expression of ryanodine receptors/sensitive stores is delayed as compared with VDCC. PMID- 9864376 TI - The cell adhesion molecule L1 is developmentally regulated in the renal epithelium and is involved in kidney branching morphogenesis. AB - We immunopurified a surface antigen specific for the collecting duct (CD) epithelium. Microsequencing of three polypeptides identified the antigen as the neuronal cell adhesion molecule L1, a member of the immunoglobulin superfamily. The kidney isoform showed a deletion of exon 3. L1 was expressed in the mesonephric duct and the metanephros throughout CD development. In the adult CD examined by electron microscopy, L1 was not expressed on intercalated cells but was restricted to CD principal cells and to the papilla tall cells. By contrast, L1 appeared late in the distal portion of the elongating nephron in the mesenchymally derived epithelium and decreased during postnatal development. Immunoblot analysis showed that expression, proteolytic cleavage, and the glycosylation pattern of L1 protein were regulated during renal development. L1 was not detected in epithelia of other organs developing by branching morphogenesis. Addition of anti-L1 antibody to kidney or lung organotypic cultures induced dysmorphogenesis of the ureteric bud epithelium but not of the lung. These results suggest a functional role for L1 in CD development in vitro. We further postulate that L1 may be involved in the guidance of developing distal tubule and in generation and maintenance of specialized cell phenotypes in CD. PMID- 9864377 TI - Activation of alphaVbeta3 on vascular cells controls recognition of prothrombin. AB - Regulation of vascular homeostasis depends upon collaboration between cells of the vessel wall and blood coagulation system. A direct interaction between integrin alphaVbeta3 on endothelial cells and smooth muscle cells and prothrombin, the pivotal proenzyme of the blood coagulation system, is demonstrated and activation of the integrin is required for receptor engagement. Evidence that prothrombin is a ligand for alphaVbeta3 on these cells include: (a) prothrombin binds to purified alphaVbeta3 via a RGD recognition specificity; (b) prothrombin supports alphaVbeta3-mediated adhesion of stimulated endothelial cells and smooth muscle cells; and (c) endothelial cells, either in suspension and in a monolayer, recognize soluble prothrombin via alphaVbeta3. alphaVbeta3 mediated cell adhesion to prothrombin, but not to fibrinogen, required activation of the receptor. Thus, the functionality of the alphaVbeta3 receptor is ligand defined, and prothrombin and fibrinogen represent activation- dependent and activation-independent ligands. Activation of alphaVbeta3 could be induced not only by model agonists, PMA and Mn2+, but also by a physiologically relevant agonist, ADP. Inhibition of protein kinase C and calpain prevented activation of alphaVbeta3 on vascular cells, suggesting that these molecules are involved in the inside-out signaling events that activate the integrin. The capacity of alphaVbeta3 to interact with prothrombin may play a significant role in the maintenance of hemostasis; and, at a general level, ligand selection by alphaVbeta3 may be controlled by the activation state of this integrin. PMID- 9864378 TI - Signal transduction via the interleukin-4 receptor and its correlation with atopy. AB - IL-4 and IL-13 are unique cytokines, in that they induce IgE synthesis in B cells and TH2 type differentiation in T cells. Both cytokines exert their biological activities by binding to their functional receptors on target cells. These receptors are thought to be composed as heterodimers, both having the IL-4R alpha chain (IL-4Ralpha) as a component. Among the signal-transducing molecules of IL-4 and IL-13, Stat6, which is activated by these cytokines and recruits to IL 4Ralpha, is essential for the biological activities of these cytokines. Atopy is an inherited tendency, underlying asthma, rhinitis, and eczema, and generating high non-specific IgE and/or high specific IgE against common antigens. Based on information on the molecular mechanism of the signal transduction of IL-4 and IL 13 and on some genetic studies, IL-4Ralpha was assumed to be one gene giving rise to atopy. One polymorphism existing in the IL-4Ralpha gene, Ile50Val, is verified to correlate with atopy by both genetic and functional aspects. On the contrary, the correlation between another polymorphism on the IL-4Ralpha gene, Arg551Gln, and atopy is still controversial. The strategy used in these studies should lead to identification of other genes involved in atopy. PMID- 9864379 TI - Translocation trisomy dup(21q) and free trisomy 21 can be distinguished by interphase-FISH. AB - The possibility of distinguishing in routine diagnostics translocation trisomy dup(21q) from disomy 21 as well as from free trisomy 21 using interphase fluorescence in situ hybridization (FISH) with a single copy probe (LSI 21) localized on chromosome 21q22.13-q22.2 is described. In free trisomy 21 and translocation trisomy dup(21q) 94%-98% of the nuclei exhibit 3 specific signals, while in disomy 21 only up to 6% of them have 3 false positive signals. Furthermore, reliable differentiation between free and translocation trisomy dup(21q) can be achieved by evaluating the percentage of nuclei with one single and two co-localized chromosome 21q22.13-q22.2 specific signals in 50-100 interphase nuclei. While in translocation trisomy 75+/-4.3% are co-localized due to a chromosomal rearrangement, in free trisomy 21 only 40+/-2.83% of the nuclei have two co-localized signals by chance. No differences in interphase signal distribution could be detected in two cases with a dicentric chromosome dup(21q) compared to one case with a monocentric one, a comparison not previously carried out. In addition, the single copy probe LSI 21 was compared with the alphoid probe D13Z1/D21Z1 which was found to be unsuitable for such assays due to polymorphisms in the a satellite regions of chromosome 21. PMID- 9864380 TI - Effects of hexose pentaacetates on electrical activity and cytosolic Ca2+ in mouse pancreatic islets. AB - Electrical activity of beta-cells and cytosolic Ca2+ concentration ([Ca2+]i) were monitored in mouse pancreatic islets exposed to the pentaacetate esters of alpha D-glucose, beta-D-galactose and beta-L-glucose, all tested at 1.7 mM concentration. In the presence of 5 mM D-glucose, alpha-D-glucose pentaacetate induced electrical activity and increased [Ca2+]i, whilst beta-D-galactose pentaacetate failed to do so. The electrical and cationic response to the D glucose ester occurred with a delay of between 5 and 10 min, the ester-induced increase in [Ca2+]i being suppressed in the absence of extracellular Ca2+. As a rule, beta-L-glucose pentaacetate also failed to evoke biophysical responses in the islets exposed to 5 mM D-glucose. However, in the presence of 10 mM L-leucine the L-glucose ester induced electrical activity. These findings, which parallel the insulinotropic action of the same esters in rat pancreatic islets reinforce the view that the positive insulinotropic action of selected hexose pentaacetates cannot be attributed to the catabolism of their acetate moiety but, instead, involves a dual mode of action linked to both the metabolism of their carbohydrate moiety and a direct effect of the ester itself upon a yet unidentified receptor system. Furthermore, this study provides the first evidence that the latter direct effect results in the induction of both electrical activity and [Ca2+]i oscillations. PMID- 9864381 TI - Regulation of triosephosphate isomerase (TPI) gene expression in TPI deficient lymphoblastoid cells. AB - The metabolic defect of triosephosphate isomerase (TPI) deficiency is reversible in deficient lymphoblastoid cells when cultured in the presence of human K562 erythroleukemia cells or plasma as exogenous source of functional enzyme. However, plasma contains a variety of undefined biological response modifiers whose effects on TPI gene expression are unknown. In the present study, TPI deficient lymphoblastoid cells were cultured in serum-free medium for 24 h (controls) and stimulated with fresh frozen plasma (FFP) at final concentrations of 20, 40, and 60% for 9 h. Changes in TPI mRNA expression were monitored by slot and Northern blot hybridisations using a specific human TPI cDNA probe. For equivalent loading of total RNA, TPI mRNA expression in FFP-treated lymphoblastoid cells exceeded that for controls by on average 20-fold. Additional studies with the transcription inhibitor, actinomycin D, revealed a rapid degradation of TPI mRNA in controls compared to FFP-treated cells, indicating that the stability of the TPI transcript was affected by plasma. These data suggest that functional or regulatory elements within the TPI gene promoter can be modulated by biological response modifiers. An understanding of the transcriptional control of TPI may provide useful insights into the development of gene therapy strategies for TPI deficiency. PMID- 9864382 TI - Effect of mutations at the residues R25, D27, P69, and N70 of B95a-MCP on receptor activities for the measles viruses Nagahata wild-type strain and CAM vaccine strain. AB - Human membrane cofactor protein (MCP, CD46) is a regulator of complement activation and also serves as a receptor for measles virus (MV). We recently isolated an MCP homolog from B95a, an Epstein-Barr virus-transformed marmoset B lymphoblastoid cell line, which is 76% identical to human-MCP. B95a-MCP acts as an MV receptor for CAM, a vaccine strain of MV, but not for Nagahata, wild-type MV strain. The four residues in human-MCP (Asp27, Lys29, Arg69, and Asp70) are reportedly MV binding sites, and these are changed in B95a-MCP (Glu27, Asp29, Pro69, and Asn70). In the present study, we constructed B95a-MCP mutants by replacing the four residues with those in human-MCP, and tested whether the Chinese hamster ovary (CHO) transfectants expressing B95a-MCP mutants become susceptible to the Nagahata strain. The CHO transfectants expressing B95a-MCP mutants formed syncytium with the CAM strain but not with the Nagahata strain. The binding of the hemagglutinin (H) of MV with B95a-MCP mutants was observed with the CAM strain but not with the Nagahata strain. These results suggest that the failure of B95a-MCP as the MV receptor for the Nagahata strain is not due simply to the natural mutations at these four residues. The critical residues for MV binding in an MCP molecule seem to differ depending upon the structure of the MV H protein. PMID- 9864384 TI - Cyclo-oxygenase-2 in vascular smooth muscle. AB - Two isoforms of cyclo-oxygenase (COX) have been identified; a constitutive isoform (COX-1), found in abundance in platelets and the vascular endothelium, considered important for the roles of prostanoids and a cytokine/mitogen inducible isoform (COX-2), which is thought responsible for the majority of the inflammatory prostanoid production. As a number of COX metabolites regulate vascular smooth muscle cell function and the interaction between the vessel and circulating components, we have discussed the possibility that COX-2 can be induced in, and regulate human arterial or venous smooth muscle cell function. It is now clear that COX-2 can be induced in freshly isolated vessels in culture, which can be further stimulated by addition of pro-inflammatory cytokines. Interestingly, smooth muscle cells derived from saphenous vein can release extremely high levels of prostanoids, and express greater levels of COX-2 protein than internal mammary artery cells. This difference can be accounted for by an arterial cell-specific negative feedback mechanism. In addition to inducing COX 2, certain cytokines regulate smooth muscle function, by regulating cell proliferation, adhesion, and mediator release. The effects of COX-2 activity on these smooth muscle cell responses will be further discussed. PMID- 9864383 TI - Transcriptional activation of CYP2C, MxA and Fas in sudden infant death syndrome. AB - The expression of cytochrome P4502C has been shown to be upregulated in sudden infant death syndrome (SIDS) and could be linked to viral infection through the release of interferon alpha and interleukins. MxA is a reliable marker of IFNalpha release and its level was significantly enhanced in SIDS reflecting the release of IFNalpha in response to viral infection. Similarly, the concentration of Fas protein was increased in SIDS (2.6x control) and indicated a stimulation of the Fas gene expression. Accumulation of MxA and Fas proteins were visible in liver and to a lesser extent in lung and kidney. The amount of RNA encoding CYP2C9 (4.4x control), 2C8 (2.5x) and 2C18 (2.3x) was markedly higher in SIDS than in age-matched children and would suggest a transcriptional activation of CYP2C gene expression. Finally, CYP2C genes were shown to be adjacent to two IFN inducible genes (IFI54 and IFI56) on chromosome 10. We conclude that in SIDS a viral infection leads to the release of IFNalpha which could activate a battery of IFN-inducible genes. This might modify the chromatin structure and facilitate the accessibility to promoter/regulatory sequences of CYP2C and Fas genes close to IFN-inducible gene on chromosome 10. PMID- 9864385 TI - Immune escape mechanisms in malignant melanoma. AB - Malignant melanoma is an antigenic tumor recognized by specific lymphocytes. Several melanoma-associated antigens have been shown to cause tumor rejection in vitro. Escape from immunosurveillance by melanoma cells is the result of several mechanisms such as total loss or decreased expression of HLA antigens, alterations in the expression of tumor-associated antigens or deficiencies in the antigen-processing machinery. Additional important features include the influence of endogenous and exogenous cytokines such as interferons or interleukins and expression of adhesion molecules or co-stimulatory molecules. All these factor have to be considered for the development of new immunoregulatory treatment modalities against advanced melanoma. PMID- 9864386 TI - Guanylin: a novel regulatory peptide possibly involved in the control of Ca2+ dependent agonist-stimulated aldosterone secretion in rats. AB - Guanylin is a 15-amino acid peptide, which activates guanylate cyclase (GC) and plays a major role in the regulation of water and electrolyte secretion by intestinal mucosa. The expression of guanylin prohormone has been recently demonstrated in the rat adrenal gland, and this prompted us to investigate whether guanylin, like other peptides secreted by adrenal medulla, affects the function of the adrenal cortex. Autoradiography demonstrated the presence of [125I]guanylin binding sites in the zona glomerulosa (ZG), but not zona fasciculata-reticularis. Guanylin did not change either basal or ACTH-stimulated steroid secretion of dispersed rat adrenocortical cells, but concentration dependently (from 10(-10) M to 10(-8) M) inhibited aldosterone response of ZG (capsular) cells to both angiotensin-II (ANG-II) and K+. Guanylin (10(-8) M) blocked the aldosterone secretagogue effect of the Ca2+-channel activator BAYK 8644, and the Ca2+-ionophore ionomycin counteracted the inhibitory action of this peptide on the secretory responses of capsular cells to ANG-II and K+. As expected, guanylin did not affect cyclic-AMP release by capsular cells, but evoked a sizeable increase in cyclic-GMP production. Both the inhibitor of GMP synthase decoyinine and the GC-inhibitor LY-83583, although suppressing cyclic GMP release, did not affect guanylin-evoked inhibition of K+-stimulated aldosterone secretion. Collectively, these findings allow us to conclude that guanylin: i) inhibits aldosterone secretion of rat ZG cells by interfering with the agonist-induced activation of voltage-gated Ca2+-channels, the stimulation of guanylate cyclase conceivably playing a negligible role; and ii) could be included in that group of regulatory peptides, secreted by medullary chromaffin cells, which are able to counteract an exceedingly high aldosterone secretion. PMID- 9864387 TI - Lovastatin induces p21WAF1/Cip1 in human vascular smooth muscle cells: influence on protein phosphorylation, cell cycle, induction of apoptosis, and growth inhibition. AB - Morbidity, mortality and the incidence of myocardial revascularisation procedures can be reduced by simvastatin, an inhibitor of the HMG-CoA reductase (EC 1.1.1.34). It was hypothesised that inhibition of isoprenylation of signalling proteins by HMG-CoA reductase inhibitors (vastatins), especially of the p21ras proteins could be causative for suppression of vascular smooth muscle cell (SMC) proliferation. The primary pharmacological mechanism of vastatins on human vascular SMC still remains unexplained. To analyse the influence of vastatins, SMC grown in presence of endothelial cell growth supplement (ECGS) were exposed to different concentrations of lovastatin. At 10 microM concentration, inhibition of SMC proliferation was associated with induction of apoptosis in a large fraction of cells as at the 1 microM level apoptosis was induced only in a minority of SMC. Protein phosphorylation on tyrosine, serine and threonine residues demonstrated no differences to untreated controls. Lovastatin induced arrest of cells in G0/G1 phase of the cell cycle and DNA synthesis was reduced. Western blot analysis demonstrated a significant induction of p21WAF1/Cip1 protein expression. This led to strong inhibition of cyclin dependent kinases (cdks) resulting in a cell cycle arrest. Our study provides evidence for a pharmacological explanation for the inhibition of ECGS-driven proliferation of human SMC by lovastatin. PMID- 9864388 TI - Opposite cellular accumulation and nitric oxide production in vivo after pleural immunization with M. leprae or M. bovis BCG. AB - Mycobacteria as intracellular pathogens have evolved mechanisms to survive within macrophages. Our previous data showed that M. leprae (ML), unlike M. bovis BCG, did not induce an inflammatory response in the mice subcutaneous tissue. Further, ML inhibited BCG-induced foot pad oedema and seemed to transform macrophages in epithelioid cells. Since these mycobacteria share common antigens, here we seeked to compare the acute and chronic cellular response evoked by ML and BCG in pleurisy of a mycobacteria-susceptible mice (BALB/c). The total leukocytes, the cell type that migrated to the pleural cavity and macrophage activation assayed by nitric oxide release were determined. Live or dead BCG Moreau recruited the same extent of cells, essentially monocytes and neutrophils, dose-dependently, in both acute and chronic pleurisy. BCG-induced eosinophilia was observed only in the acute response (after 24 h of injection). A significant nitric oxide release by pleural macrophages was triggered by BCG Moreau without previous activation. Nevertheless, ML failed to recruit leukocytes to the pleural space or to lead to nitric oxide production despite the number of bacilli used and the time studied (1, 7 or 14 days after injection). Although these mycobacteria have common antigens that cross-react, these data show a distinct ability of ML or BCG to recruit cells to the pleural space and to activate pleural macrophage for nitric oxide production in vivo. PMID- 9864389 TI - Assessment of genomic imbalances in malignant fibrous histiocytomas by comparative genomic hybridization. AB - In order to investigate genomic imbalances, comparative genomic hybridization was applied to 20 malignant fibrous histiocytomas. Deletions were rare and found mainly in chromosomes 2q33-35, 4q32-qter, 8p, 9p21-pter, 12p and 19p, whereas, over-representations frequently affected chromosomes 3, 4q31, 5p, 6, 7, 14q22 ter, 18p, as well as, five distinct amplifications within the regions 12q12-15 and 15q24-qter. The total number of genetic imbalances per tumor was slightly increased in primary tumors when compared to relapses. No relationship was found between the patterns of gain and loss when compared to the histological subtype, tumor grading, the clinical outcome and the p53 mutation status. PMID- 9864390 TI - Gene therapy for gliomas: p53 and E2F-1 proteins and the target of apoptosis. AB - Current therapy for glioma is suboptimal. The transfer of apoptosis genes to tumors constitutes one of the most promising strategies for cancer gene therapy. We have previously shown that massive apoptosis occurs when wild-type p53 or E2F 1 expression is induced in glioma. However, the mechanism of action and the efficiency in inducing apoptosis of these two proteins are not similar. Adenovirus-mediated p53 gene transfer is ineffective in causing apoptosis in glioma cells that retain wild-type p53 genotype or overexpress the p21 protein. The p16/Rb/E2F pathway is the most frequent target of genetic alterations in gliomas, and therefore constitutes a suitable target for gene therapy strategies. However, the transfer of either the p16 or Rb gene to glioma cells results in cytostatic effect. The E2F-1 protein is able to induce generalized apoptosis in gliomas independently of the p53, p16 or Rb status. In addition, p21- or p16 mediated growth arrest did not protect glioma cells from E2F-1-mediated apoptosis. The apoptotic molecule bax is induced in p53-mediated apoptosis, but bax is not induced in E2F-1-mediated apoptosis in glioma cells. Careful selection of patients may be necessary before designing therapeutic strategies using either p53 or E2F-1 as a therapeutic tools for glioma patients. PMID- 9864391 TI - Enhancement of XPG mRNA transcription by human interferon-beta in Cockayne syndrome cells with complementation group B. AB - Human interferon-beta (HuIFN-beta) confers UV-refractoriness in association with increased DNA repair capacity to human cells. We examined the modulation of XPG gene expression by HuIFN-beta in UV-sensitive cells from Cockayne syndrome complementation B (CSB), xeroderma pigmentosum complementation A (XPA) and normal control cells. Northern blot analysis revealed that XPG mRNA was more extensively transcribed in CSB cells treated with HuIFN-beta than in those without HuIFN-beta treatment. XPG mRNA from XPA cells and normal control cells was not markedly transcribed by HuIFN-beta treatment compared to that from CSB cells. The findings suggested that different mechanisms of UV-refractoriness by HuIFN-beta exist between CS and XP. PMID- 9864392 TI - Host cell-dependent replication of HIV-1 gag MA, CA, and NC mutants are independent of the functions of Vif and Vpu. AB - We have previously shown that some gag gene mutants of human immunodeficiency virus type 1 (HIV-1) display a replication-defect in a cell-dependent manner. We and others have also demonstrated that the requirement of vif and vpu genes for HIV-1 replication is cell-dependent. To determine whether the cell-dependent growth of the HIV-1 gag mutants is related to the functions of Vif and Vpu, double mutants of gag-vif and gag-vpu were constructed, and monitored for their replication in various cell lines. The results obtained showed that the mutations in gag do not affect the cell-dependent functions of Vif and Vpu. PMID- 9864393 TI - An effective tumor vaccine against malignant melanoma: irradiated autologous tumor cells admixed with MVE-2. AB - The aim of this study was to develop as effective as possible autologous tumor vaccine which would be at the same time easy to produce, highly controllable, and without undesired side effects on normal tissue. Therefore, irradiated autologous - syngeneic B-16 tumor cells admixed with a non-specific immunomodulator MVE-2 (a polymer fraction of 1,2-co-polymer of divinyl ether and maleic anhydride) were used for subcutaneous (s.c.). or intraperitoneal (i.p.) prevaccination of experimental mice. Compared to the control mice, a statistically significant delay in tumor development of s.c. tumors was achieved in prevaccinated mice (p<0.05). An even better effect was observed in mice challenged i.p. with viable tumor cells. Using a single prevaccination complete protection was obtained in between 40-85% of the experimental mice. When the survivors from the groups injected once with the tumor vaccine were rechallenged with viable tumor cells (101 day after the first tumor challenge, no additional prevaccination), 15.7% remained free of tumor, while the survivors from the groups injected with the tumor vaccine twice and 101 day later rechallenged with viable tumor cells remained free of tumor in 60% of the cases. Based on these results we can postulate that our vaccine is effective for prevention of tumor development. The achieved protection can be augmented with serial vaccinations and can be maintained for a longer period of time. PMID- 9864394 TI - Human chorionic gonadotropin as a target for cancer vaccines. AB - Human chorionic gonadotropin (hCG) is expressed by common cancers and may play a role in cell transformation as well as angiogenic, metastatic, and immune escape phenomena that are central to cancer progression. Clinical trials with a vaccine targeting the carboxy-terminal peptide of -hCG have indicated that tolerance to this oncofetal antigen can be broken. Humoral responses that may modulate the biologic activity of tumor-associated hCG as well as cellular responses to hCG have been generated. Studies are in progress to further define the biologic significance of hCG in cancer and to develop a vaccine approach that will best target this expression. PMID- 9864395 TI - Clinical significance of p53 in non-small-cell lung cancer. AB - p53 abnormalities appear to play critical roles in human carcinogenesis. Many studies have indicated that p53 alteration is responsible for the differences in clinical characteristics of non-small cell lung cancer (NSCLC). The purpose of this review is to evaluate and summarize the significance of p53 alteration in the diagnosis and therapy of NSCLC. After assessing the principles of each approach to detecting p53 alteration, we reviewed the literature describing the impact of p53 alteration on clinical modalities such as diagnosis, prediction of prognosis, and response to treatment. We also reviewed p53 gene replacement therapy for NSCLC. Although p53 appears to be the most important gene in the development of NSCLC, it is not known whether p53 abnormalities are useful markers for clinicians. A multicenter, prospective study of p53 immunohistochemistry should therefore be performed. Replacement of the p53 gene has been shown to be effective in a clinical trial for patients with NSCLC. In addition to simple gene replacement, combination treatments of gene transfer and chemotherapy are now being tested in clinical trials. PMID- 9864396 TI - Post-transplantation lymphoproliferative disorder of the T-cell/B-cell type: an unusual manifestation in a renal allograft. AB - Marked impairment of the cellular immune system predisposes renal transplant recipients to Epstein-Barr virus (EBV) associated clinical syndromes. This can culminate in post-transplantation lymphoproliferative disorders (PTLD) and malignant lymphomas. An unusual PTLD in a 59-year-old renal transplant recipient is reported here. Sonography and CT scan revealed a hypovascular infiltrating tumor mass in the lower pole of the graft which on histopathologic examination revealed a monotonous lymphoid proliferation. T-cell receptor and immunoglobulin heavy chain gene rearrangement as well as immunohistochemical analyses demonstrated a polyclonal origin of atypical lymphatic T- and B-cells. The Epstein-Barr viral genome was detected in the mass by Southern blot analysis, and a primary EBV infection was confirmed by serologic studies. Clinical follow-up showed a tumor-free course till the patient's sudden cardiac death 14 months after the operation. PMID- 9864397 TI - Alteration of caspase-3 (CPP32/Yama/apopain) in wild-type MCF-7, breast cancer cells. AB - Caspase-3 (CPP32/Yama/apopain), one of the interleukin 1 -converting enzyme (ICE) like proteases (caspases), is anticipated to mediate apoptotic cell death. We observed the expression of caspase-3 in various cancer cell lines and lack of normal expression of mRNA and protein in MCF-7, human breast carcinoma cell line. Sequence analysis of cDNA showed 125 nucleotides deletion in spite of no gross gene alteration of caspase-3 in MCF-7. The possible cause is altered splicing of the fragment followed by frame shift at translation level. MCF-7 cells are widely used in the research of apoptosis because of the high sensitivity to tumor necrosis factor induced cell death. However, our results suggest the existence of other apoptotic pathways independent on caspase-3 at least in MCF-7 cells. PMID- 9864398 TI - Cell-killing by paclitaxel in a metastatic murine melanoma cell line is mediated by extensive telomere erosion with no decrease in telomerase activity. AB - The purpose of this study was to investigate and compare the effects of paclitaxel and its water-soluble conjugates (sodium-pentetic acid-paclitaxel; polyethylene glycol-paclitaxel, and poly[L-glutamic acid]-paclitaxel) on chromosome morphology and induction of apoptosis in a metastatic murine melanoma cell line (K1735 clone X-21). For this, murine melanoma cells were treated continuously for 72 h with three concentrations (1.2 microM, 2.4 microM, and 4.8 microM) of each of paclitaxel, and conjugates. Another set of cells were pulse treated at 2.4 microM, 4.8 microM and 9.6 microM concentrations of each of these drugs for 4 h and the recovered cells were examined after 72 h. Control cultures received only the solvents (dimethyl sulfoxide or water). Our results showed a significant increase in the frequencies of telomeric associations, chromosome aberrations, polyploidization, distorted and disintegrated chromosome morphology, and reduced telomeric signal intensity by fluorescence in situ hybridization, in treated cultures as compared to the controls. However, we detected no change in telomerase activity. In addition, the majority of interphase nuclei in treated cells showed apoptotic bodies, with chromatin condensation. These in vitro results suggest that cell death induced by paclitaxel and its water-soluble conjugates is due to the loss of telomeric repeats, as shown by reduced signal flourescence and increased telomeric associations. PMID- 9864399 TI - 99m Tc MIBI prone scintimammography in breast Paget's disease: a case report. AB - A 99m Tc MIBI prone scintimammography (PSM) was performed in a case of underlying Paget's disease of the breast. 99m Tc MIBI PSM showed a diffuse scintigraphic image like a spread of uptake from the deeply located zones of the breast toward epidermis. In vivo, 99m Tc MIBI PSM represents the spread of neoplastic Paget's cells probably attracted by chemotactic factors released by keratinocytes. This spread in Paget's disease is correlated to neu oncogene overexpression which increases the metastatic activity as a consequence of motility enhancement and growth stimulation effect. These scintigraphic images suggest that 99m Tc MIBI PSM could be relevant in management of Paget's disease of the breast. PMID- 9864400 TI - Effects of interleukin-2 transduction on the human hepatoma cell lines using retroviral vector. AB - Gene therapy, using cytokine gene transduction, aims to increase the antigenicity of tumor cells, and to activate the immune effector cells, and thereby inducing tumor regression. With regards to in vitro sensitivity to peripheral blood monocytes and in vivo tumorigenic activity we compared the differences between parent hepatoma cell lines and interleukin-2 (IL-2) transduced hepatoma cell lines using N2A/IL-2 and LNC/IL-2 retrovirus. IL-2 secretion was 186 pg/10(6) cells/24 h in SK-Hep1 cell line and 147 pg/106 cells/24 h in Hep-3B cell line with N2A/IL-2 retroviral vector and was 55,000 pg/10(6) cells/24 h in Hep-3B cell line with LNC/IL-2 retroviral vector. in vitro sensitivity to peripheral blood monocytes was increased by 163.8-254% in IL-2 transduced hepatoma cell lines (Hep 3B/LNC/IL-2, Hep-G2/LNC/IL-2) compared to those of the parent cell lines. The tumor was formed in 1 of 3 BALB/c mice and all 3 nude mice with the injection of 1x107 cells. Simultaneous injection of 1x10(7) cells of the parent cell line (Hep 3B) into the right flank and IL-2 transduced cell line (Hep-3B/LNC/IL-2) into the left flank of the three BALB/c mice and of 5x10(5) cells for the three nude mice resulted in a complete regression of the IL-2 modified tumor cell line (Hep 3B/LNC/IL-2) in 3 weeks and the parent cell line (Hep-3B) in 5 weeks. After injection of 1x10(7) cells into five other nude mice, the tumor of the IL-2 transduced hepatoma cells (Hep-3B/LNC/IL-2) gradually disappeared, however, the tumor of the parent hepatoma cell line initially decreased and then gradually regrew 20 days later. In conclusion, IL-2 transduced hepatoma cell lines secreting IL-2 became more sensitive to peripheral blood monocytes. IL-2 secretion by LNC/IL-2 retrovirus from the hepatoma cell lines was more prominent compared with that by N2A/IL-2 retrovirus. IL-2 transduction into the hepatoma cells resulted in increased antigenicity to the tumors formed by IL-2 transduced hepatoma cell line and parent cell line, which leads the regression of the tumors. However, the higher the tumor burden, the less efficient tumor regression by IL-2 transduction into the hepatoma cell line in nude mice was observed. PMID- 9864401 TI - Scrotal ulcer occurring in patients with acute promyelocytic leukemia during treatment with all-trans retinoic acid. AB - Although all-trans retinoic acid (ATRA) has been shown to improve the outcome of patients with acute promyelocytic leukemia (APL) compared with chemotherapy alone, various adverse effects have been reported. We report here the development of scrotal ulcer in four patients with APL during ATRA treatment. ATRA 45 mg/m2 was administered orally to four patients with newly diagnosed APL, two of whom also received chemotherapy. Scrotal ulcers appeared in all four patients after a median of 22 days (range: 17-29 days) of ATRA treatment. ATRA was discontinued in only one patient, who was then treated with methylprednisolone pulse therapy. The fever resolved, and scrotal ulcer improved after this treatment. The other three patients were treated with steroid or antibiotic ointment for scrotal ulcers. ATRA was re-administered 2 years later in one patient who had relapse of APL. Ulcers appeared again on the scrotal and lower abdominal skin on day 27 of ATRA treatment. These findings strongly suggest that scrotal ulcer is a potential adverse effect of ATRA. PMID- 9864402 TI - Increased chromosome exchange frequencies in iodo-deoxyuridine-sensitized human SW-1573 cells after gamma-irradiation. AB - The induction of chromosome exchanges was investigated in SW-1573 human lung tumour cells radiosensitized with iododeoxyuridine (IdUrd) and irradiated with gamma-rays. Following treatment chromosome 2 and X were analyzed using fluorescence in situ hybridization (FISH) with chromosome-specific DNA libraries. The yield of chromosome exchanges involving chromosome 2 was higher than those involving chromosome-X. On the basis of the DNA content the relative involvement of the X-chromosome in exchange frequencies after 2 Gy was much higher than of chromosome 2. After 4 Gy the relative involvement of both chromosomes in exchanges is approximately equal. After radiosensitization, increased chromosome exchange frequencies are observed in both studied chromosomes. For the total chromosome exchange frequencies the sensitizer enhancement ratio (SER) at 2 Gy is 1.8 and 1.3 for chromosome 2 and X respectively. The SER at 4 Gy for total exchange frequencies is 1.6 and 1.9 chromosome 2 and X respectively. For reciprocal exchanges at 2 Gy higher SER values and at 4 Gy lower SER values were observed for both chromosomes. PMID- 9864403 TI - Cytokines in human breast cancer: IL-1alpha and IL-1beta expression. AB - We hypothesize that interleukin 1alpha (IL-1alpha) and interleukin 1beta (IL 1beta) are present and tumor cell associated in human breast cancer (HBC) specimens. To test our hypothesis: a) immunologic analysis was performed on HBC histologic sections for IL-1alpha (n=49) and IL-1beta (n=42) distribution; and b) homogenates of HBC tumors were analyzed for levels of IL-1alpha (n=82), IL-1beta (n=101) and interleukin 8 (IL-8) (n=103) expression. Immunohistochemical analysis demonstrated the presence of IL-1alpha and IL-1beta in tumor cells in patients with invasive cancer and ductal carcinoma in situ. Quantitative analysis confirmed the presence and positive correlation of IL-1alpha and IL-1beta to IL 8, a known angiogenic factor, in cancer specimens. These studies demonstrate that tumor-associated IL-1alpha+, IL-1beta are present in the tumor microenvironment and may play a pivotal role in regulating breast tumor growth and metastasis. PMID- 9864404 TI - Prospective study of nonsteroidal anti-inflammatory drugs and breast cancer. AB - We conducted a prospective cohort study of Nonsteroidal anti-inflammatory drugs (NSAIDs) and breast cancer among 32,505 women in central Ohio, USA. After 5 years of follow-up, a total of 393 cases have been detected. The annual incidence of breast cancer per 100,00 women varied inversely with increasing intake of NSAIDs, declining from 323 among non-users to 183 among heavy users (p<0.01). Breast cancer rates decreased by about 50% with regular ibuprofen intake (p<0.01), and by about 40% with regular aspirin intake (p<0.05). The results suggest that specific NSAIDs may be effective chemopreventive agents against breast cancer. PMID- 9864406 TI - The reduced expression of e-cadherin, alpha-catenin and gamma-catenin but not beta-catenin in human lung cancer. AB - Cadherins are Ca2+-dependent cell-cell adhesion molecules, and are involved in the formation and maintenance of the histo-architecture. Using a combination of biochemical and immunohistochemical methods, we analyzed the expression of cadherin-catenin complexes in 37 non-small cell lung carcinomas. In 19 cases, decreased expression of E-cadherin protein was observed. In 12 of them, decreased expression of alpha-catenin protein was also observed. Thus, decreased expression of alpha-catenin was apparently preceded by decreased expression of E-cadherin. In no cases was decreased expression of beta-catenin observed. In the 12 cases in which mRNA expression was analyzed by Northern blot analysis, decreased expression of mRNAs for E-cadherin and alpha-catenin was observed in 11 and 9 cases, respectively. In cases with reduced E-cadherin and alpha-catenin expression, immunohistochemistry revealed two types of staining pattern for the proteins. In the first type, almost all the cells in a tumor were stained weakly (homogeneous pattern). In the second type, different percentages of cells were stained strongly, the rest being almost negative for the staining (heterogeneous pattern). PMID- 9864405 TI - Synchronous primary lung cancer presenting with small cell carcinoma and non small cell carcinoma: diagnosis and treatment. AB - Synchronous primary lung cancer (SPLC) occurs in up to 0.5% of patients with lung cancer. Among SPLC cases, coexistence of small cell carcinoma (SCLC) and non small cell carcinoma has been reported in a very small fraction. Futhermore, there have been no reports discussing treatment and prognosis of SPLC presenting with SCLC and NSCLC. We report on two cases of SPLC presenting SCLC in limited stage and operable NSCLC. One patient developed synchronously SCLC and adenocarcinoma of the lung, while the other SCLC and squamous cell carcinoma of the lung. The clonal origin of these synchronous lung cancers was evaluated using immunohistochemical and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analyses. Both of the patients were diagnosed based on transbronchial lung biopsy (TBLB) and mediastinoscopic biopsy. They were successfully treated with chemoradiotherapy and adjuvant surgery, and are now doing well without any signs of tumor progression for about one year. In both cases, a response of mediastinal lymph node for concurrent chemoradiotherapy was quite different from that of the mass in the lung field. In case 2, p53 mutation was observed in the SCLC tissue, but not in the NSCLC tissue by PCR-SSCP. In both cases, carcinoembryonic antigen was documented in the NSCLC tissue, but not in the SCLC tissue by immunohistochemical staining. This report indicates the importance of the accurate diagnosis of SPLC by employing TBLB and/or media stinoscopy for the optimal treatment of patients having SPLC presenting with SCLC and NSCLC. Diagnostic criteria and standard treatment of this disease should be established. PMID- 9864408 TI - Cyclin E overexpression in human gallbladder carcinomas. AB - The expression of cyclin E, one of the important positive cell cycle regulators, was examined immunohistochemically in gallbladder carcinomas. Cyclin E gene product was detected in 58 (49%) out of 118 cases. The degree of cyclin E expression was not associated with any clinicopathological factor including histology, the depth of tumor invasion, tumor stage and patient prognosis. Cyclin E expression was not correlated with that of p53 protein statistically, whereas it was correlated with the proliferative activity of the tumor cells by PCNA (p<0.05). These results suggested that cyclin E expression may confer progression of gallbladder carcinomas. PMID- 9864407 TI - Fibroblast growth factor-9 (glia-activating factor) stimulates proliferation and production of glial fibrillary acidic protein in human gliomas either in the presence or in the absence of the endogenous growth factor expression. AB - We tested fibroblast growth factor-9 (FGF-9) expression in human glioma cells (U251MG, T98G, U87MG, KALS-1, NMC-G1) and only NMC-G1 expressed endogenous FGF-9. All cells expressed bFGF and high affinity receptors for FGFs (FGFR1 and FGFR3). Exogenously supplied bFGF and FGF-9 both showed mitogenic activities in all cells. Neutralizing antibody against bFGF inhibited the proliferation in U251MG and NMC-G1, however that against FGF-9 inhibited the proliferation only in NMC G1. GFAP expression was stimulated by both FGFs in these cells. FGF-9 potentially regulates proliferation and GFAP expression in human gliomas either in the presence or in the absence of the endogenous growth factor expression. PMID- 9864409 TI - Comparison of p53 and CD44 variant 6 expression between paired primary and recurrent ovarian cancer: an immunohistochemical analysis. AB - Progression of malignant potential in ovarian cancer was investigated by comparing recurrent tumor with their primary tumor counterparts in terms of p53 and CD44 variant 6 (CD44v6) expression. Forty-three paired primary and recurrent tumors were immunohistochemically evaluated for expression of p53 and CD44v6. A paired analysis revealed that p53 and CD44v6 expression were significantly greater in recurrent tumors than those in the primary counterparts with statistical significance, respectively (P=0.0055 and 0.0071; Wilcoxon signed-rank test). No relationship between these two proteins was found. These results suggested that recurrent ovarian cancer may possibly express a stronger malignant potential in comparison to the primary tumor. PMID- 9864410 TI - Management of primary gastric lymphomas from a surgeon's viewpoint. AB - Primary gastric lymphoma is a relatively uncommon disease and controversy still exists over its management. In order to assess prognostic factors of lymphomas, we carried out a retrospective study of 28 surgically treated gastric lymphoma patients. The overall survival rate for all the patients in the study was 33% at 10 years. On univariate analysis, lymph node metastasis and depth of tumor infiltration proved to be a significant prognostic factor while size and location of tumor, sex, and method of resection was not. From our results we believe that gastric lymphoma can be regarded as a localized disease in the early stages and a curative resection can be attained when aggressive surgery is possible. PMID- 9864411 TI - A polyclonal antibody against synthetic peptide conserved in N-Myc protein reacts with water-soluble recombinant N-Myc protein. AB - The importance of determining the N-Myc protein has been emphasized in neuroblastoma. We attempted to obtain a water-soluble N-Myc protein, and an antibody highly specific for the N-Myc protein. A plasmid was constructed from partial exons 2 and 3 of the N-myc gene, and expressed in Escherichia coli by isopropyl-beta,-D-thiogalactopyranoside. Newly obtained N-Myc (rN-Myc) was water soluble with a M.W. of 38 kDa. An N-Myc-specific peptide, GVAPPRPGG RQTSGGDH, was used to raise an antibody. Specificity of the obtained antibody was confirmed and we found the rN-Myc protein reacts positively with the anti-N-Myc IgG raised here. The rN-Myc protein and the anti-N-Myc IgG obtained are expected to be used in an ELISA for N-Myc. PMID- 9864412 TI - High expression of glycosphingolipids involved in procoagulant activity of cancer cells. AB - To investigate the involvement of tumor-associated antigens such as sLe(a) and sLe(x), as well as Le(y) glycosphingolipid in the procoagulant activity of cancer cells, we examined the relationship between the inhibitory effect of anti-sLe(a) or anti-sLe(x) antibodies on the procoagulant activity of cancer cells, and cell surface expression of sLe(a) and sLe(x) antigens. We observed that the procoagulant activity of cancer cells which highly expressed sLe(a) or sLe(x) antigens (expression rate >70%) was significantly inhibited by the relevant antibodies. These data suggested that not only Le(y) but also sLe(a) and sLe(x) antigens play an important role in coagulation induced by various cancer cells. PMID- 9864413 TI - Replication error in human breast cancer: comparison with clinical variables and family history of cancer. AB - Replication errors (RER) at microsatellite repeats indicate genomic instability in hereditary nonpolyposis colorectal cancer (HNPCC) and in some sporadic cancers. We have studied genomic instability in 313 sporadic breast tumors and in 106 tumors from BRCA2, 999del5 carriers at 43 genomic loci on 13 chromosomes. RER was observed in 8/419 (1.9%) of the cases at one or more chromosomal loci. The frequencies of type I and type II RER were similar. The majority of RER+ tumors showed ER+, PgR+, high S-phase fraction, tumor size >2 cm and LOH at 2p, 2q and 3p. All 8 RER+ tumors were of the ductal histotype. The breast cancer cases with RER are not part of an HNPCC syndrome and a family history of colorectal cancer growth is not detected in relatives, with the exception of one case. However, four of the RER+ cases are from individuals carrying the BRCA2, 999del5 mutation. We conclude that RER is a rare somatic event during human breast carcinogenesis and may be associated with progression of breast carcinomas. PMID- 9864414 TI - Stage II vaginal cancer responding to chemotherapy with irinotecan and cisplatin: a case report. AB - We report a patient with stage II primary vaginal cancer whose tumor disappeared on magnetic resonance imaging after a single course of neoajuvant chemotherapy using irinotecan and cisplatin. The regimen was 60 mg/m2 of cisplatin (day 1) and 60 mg/m2 of irinotecan (days 1, 8, 15). After addition of 1 course of cisplatin alone, radical surgery was performed. Histological examination of the excised tissue revealed no cancer cells. This is the first report on the good efficacy of chemotherapy with irinotecan and cisplatin for advanced vaginal cancer. PMID- 9864415 TI - Thallium-201-chloride and technetium-99m-MIBI SPECT of primary and metastatic lung carcinoma. AB - We compared technetium-99m-MIBI (Tc-99m MIBI) with thallium-201-chloride (Tl-201) SPECT imaging in patients with lung carcinoma. In addition, we compared the imaging characteristics of Tl-201 and Tc-99m MIBI after radiation therapy. Thirty seven patients with primary lung carcinoma were evaluated with SPECT imaging for metastasis to the mediastinal lymph nodes and brain. Patients were imaged with Tl 201 chloride images 10 and 180 min after injection. Patients were also imaged 10 min after injection of Tc-99m MIBI. The sensitivity of Tl-201 SPECT for the primary lesion and brain metastasis was 97.1% and 70.0% respectively at 10 min and 97.1% and 60.0% at 180 min. The sensitivity of Tc-99m MIBI SPECT was 97.1% (for the identification of the primary lesion) and 50.0% (for the detection of brain metastasis) at 10 min. The uptake ratios (count in tumor/count in normal lung or brain) at 10 min on the Tl-201 SPECT and on the Tc-99m MIBI SPECT were not significantly different for the primary tumor or for brain metastasis. The uptake ratios were better for Tc-99m MIBI than for Tl-201 [2.82 vs. 1.99 (p<0.05)] for mediastinal lymph nodes. Decreasing uptake ratios and retention index with both agents after radiation therapy are concordant to the follow-up clinical course. Tc-99m MIBI SPECT is more sensitive in the detection of metastasis to mediastinal lymph nodes than Tl-201 SPECT. PMID- 9864416 TI - Surgical margin and breast recurrence after breast-conserving therapy. AB - A total of 161 patients with clinical stage I and II breast cancer received breast-conserving therapy between August 1991 and December 1997, and local recurrence occurred in five patients. The actuarial local control 5 years after breast-conserving surgery was 96.6%. We studied microscopic surgical margins of resected specimens in patients with breast-conserving surgery to determine whether the surgical margin was a risk factor for local recurrence in the conserved breast. Microscopic margins were negative in 125 (78%) of 161 patients and positive in 36 (22%). There were no differences between patients with positive surgical margins and those with negative surgical margins in age at operation, tumor size, clinical stage, lymph node status, estrogen receptor status, or distance from tumor to nipple. Local control was significantly better in the surgical margin-negative patients than in the surgical margin-positive patients. We conclude that microscopic surgical margin is a risk factor for local recurrence in the conserved breast. PMID- 9864417 TI - A case-control study of the relationship between dietary factors and risk of lung cancer in women of Shenyang, China. AB - A 1:1 case-control study of lung cancer incidence among women in Shenyang was conducted to explore the relationship between diet and the risk of lung cancer, with emphasis on the potential effects of a list of specific dietary constituents on modifying lung cancer risks. Dietary information on 290 cases and population matched controls was obtained by personal interviews. Multiple logistic regression models were used for the statistical analysis and summarization of the data. A significant difference was found between cases and controls with respect to intake of beta-carotene, vitamin C and fibers, all of which reduced the risk for lung cancer in a dose-dependent manner; yielding calculated odds ratio (OR) of 0.84, 0.75, and 0.46, respectively. The apparent effects of these nutrients persisted after adjusting for cigarette smoking;suggesting that they may function as protective factors to reduce the risk for lung cancer in Chinese women. PMID- 9864418 TI - Detection of UV-induced K-ras codon 12 mutation by PCR and differential dot-blot hybridization in cells from Down syndrome and Cockayne syndrome. AB - By means of the polymerase chain reaction (PCR) and differential dot-blot hybridization, base substitution mutations of K-ras codon 12 were investigated in skin fibroblast cells from Down syndrome (DS) patients. Mutations were identified in DS cells after UV irradiation, predominantly in cells from younger patients. In contrast, no mutation was detected in cells from Cockayne syndrome (CS) patients who had the same features of premature aging as in DS but were not prone to cancer. This association of DS cells, but not CS cells, with inducibility of the K-ras codon 12 mutation may imply the proneness of DS patients to cancer development but a lack of proneness of CS patients. PMID- 9864419 TI - Expression of type I interferon receptor and its relation with other prognostic factors in human neuroblastoma. AB - Expression of type I interferon receptor (IFN-R) has been found in several normal tissues and in malignant neoplasms, mainly those with epithelial differentiation. In order to analyze the immunohistochemical expression of type I IFN-R we studied 79 cases of neuroblastoma. Results of expression of type I IFN-R were statistically correlated with histopathology, stage, bcl-2 and PCNA expression, N myc amplification and apoptosis. We found expression of type I IFN-R in 54/79 cases showing statistical correlation with bcl-2 expression (P=0.017) and favourable histopathology (P=0.015). The overexpression found in ganglion cells suggests that IFN-R could be involved in the pathway of neuroblastoma differentiation. Moreover, the expression of type I IFN-R in stage 4 cases (12/20), even with N-myc amplification (6/8), opens new possibilities for therapeutic management in advanced cases that do not respond to any chemotherapeutic protocol. PMID- 9864420 TI - The measurement of cadmium in biological materials, using graphite furnace atomic absorption spectrometry with Zeeman background correction. AB - Cadmium (Cd), a toxic heavy metal and probable carcinogen in humans, appears to have potential anti-cancer activity in pre-clinical systems. This observation led us to develop a method for measuring cellular Cd and DNA-bound Cd following micromolar exposures to cadmium dichloride. Cultured human ovarian cancer cell lines were used. Following low level exposures to cadmium dichloride (CdCl2), atomic absorption spectrometry with Zeeman background correction was used to measure total cell associated Cd in wet-ashed cells, and the lower limits of detection was at 100 pg of Cd per 106 cells. In cellular DNA isolated by cesium chloride density gradient centrifugation, levels of 1.5 Cd lesions (Cd molecules) per 106 nucleotides were reproducibly detected. Standard curves with samples yielded 76.4 6.7% recovery when using picogram quantities of Cd. Manipulation of the total amount of biological material used, can further improve detection limits. Thus, this method is suitable for the detection of Cd in biological matrixes after low levels of Cd exposure, and shows good performance in terms of the level of sensitivity and reproducibility. PMID- 9864421 TI - Penclomedine-induced DNA fragmentation and p53 accumulation correlate with reproductive cell death in colorectal carcinoma cells with altered p53 status. AB - Penclomedine, a synthetic pyridine derivative, has documented antitumor activity and is being investigated in clinical trials. Its mechanism of action is unknown although it may be metabolized to a free radical, DNA-reactive species. We previously reported that telomerase positive colorectal carcinoma (RKO) cells with abrogated p53 function were more sensitive to penclomedine than were telomerase positive cells with wild-type p53. The present study demonstrates that significant differences in DNA fragmentation in response to penclomedine were observed in RKO cells lacking functional p53 compared with RKO cells with normal p53 function. No differences in DNA fragmentation in response to ionizing radiation were seen in RKO cells with normal or abrogated p53 function. RKO cells with functional p53 respond to penclomedine treatment with a dose-dependent increase in p53 protein levels. However, RKO cells with abrogated p53 function did not show any such change in p53 protein levels. Further, p53-independent increase of p21 was observed, although the significance of this response remains uncertain. These studies suggest that penclomedine may have a therapeutic advantage in killing cells that have abrogated p53 function. PMID- 9864422 TI - Comparative p53 mutational spectra in aerodigestive tract tumors in Hong Kong Chinese patients. AB - The p53 mutations in colorectal, esophageal and lung carcinomas from Hong Kong were studied previously. The availability of mutation data for these cancers in one geographical region prompted investigations into special features of these p53 alterations. Mutations in codons 175, 176, 248 and 273 accounted for 35% of all mutations detected. A hot spot at codon 176 observed in esophageal carcinomas was not detected in any of the other aerodigestive tract tumors studied and appeared to be uniquely restricted to Chinese esophageal cancers. Other unique mutation sites, and a notably higher frequency of insertions and deletions in each of these cancers, were also detected in Chinese patients as compared to Caucasians. PMID- 9864423 TI - Toxicity and antitumor activity of interferon gamma alone and in combinations with TNFalpha and melphalan in isolated limb perfusion in the BN175 sarcoma tumor model in rats. AB - Isolated limb perfusion (ILP) with TNFalpha, melphalan (M), and IFNgamma results in high tumor response rates in patients with soft tissue sarcomas, melanomas and other tumors. IFNgamma can act synergistically in combination with TNFalpha but in clinical studies this has not been fully investigated. In the BN175 rat sarcoma limb perfusion model we investigated the role of IFNgamma. There were 8 different treatment groups: (I) sham ILP (n=9); (II) IFNgamma alone (n=10); (III) TNFalpha 50 microg (n=9); (IV) TNFalpha + IFNgamma (n=6); (V) melphalan (M) 40 microg (n=11); (VI) M + IFNgamma (n=6); (VII) TNFalpha + M (n=27); (VIII) TNFalpha + M + IFNgamma (n=9). Tumor response and hindlimb function were analyzed. In group I-VI no tumor regressions were observed at 5 days after ILP. ILP with TNFalpha + M had highly effective response rate (RR) of 73%; complete response (CR) rate 55%), very similar to RR in patients. Addition of IFNgamma increased the RR by 16% to 89% and the CR rate by 23% to 78%. This difference was not statistically significant. When IFNgamma was added to TNFalpha or TNFalpha + M it increased limb toxicity significantly (p<0.05 and p<0.005). Since such regional toxicity has not been observed in patients while similar increases in tumor response rates have been reported with IFNgamma it is of importance to define the role of IFNgamma in the clinical setting. PMID- 9864424 TI - Radioimmunotherapy of orthotopically transplanted pancreatic cancer with 131I labeled chimeric monoclonal antibody Nd2. AB - Recombinant mouse/human chimeric antibody Nd2 was previously reported to have the same specificity and reactivity against pancreatic cancer as murine Nd2 without HAMA production which induced serious problems in clinical application. We established orthotopic SW1990 transplanted mice to represent the progression of pancreatic cancer. We examined anti-tumor effect of 131I-labeled chimeric Nd2 in an orthotopic transplant model. 131I-labeled chimeric Nd2 inhibited the growth of orthotopically transplanted tumors and extended the survival time of the mice. These results suggest that 131I-labeled chimeric Nd2 has high clinical applicability in the therapy of pancreatic cancer. PMID- 9864425 TI - Bone scan and bone biopsy in the detection of skeletal metastases. AB - This study provides the first quantitative indication of the limits of sensitivity of a bone scan with technetium (99Tc-MDP) in detecting skeletal metastases and thereby also helps to explain the fact that bone scans may be negative when metastases are present in the bone marrow. Since 99Tc-MDP remains the least noxious and most widely used isotope for bone scanning, these results have direct clinical relevance in the evaluation of patients with solid tumors and possible metastatic spread. PMID- 9864426 TI - Spontaneous pelvic hematoma simulating neoplasm: case report and literature review. AB - Spontaneous hematomas are rare and can present with acute or chronic symptoms. Our patient presented with deep vein thrombosis of the lower extremity associated with a spontaneous pelvic hematoma. This lesion was radiologically and clinically indistinguishable from a soft-tissue neoplasm. The case of a spontaneous pelvic neoplasm in an otherwise healthy young man is presented and the literature reviewed regarding issues of differential diagnosis and clinical management. PMID- 9864427 TI - Polymorphisms of the p53 gene in women with ovarian or endometrial carcinoma. AB - The p53 gene is frequently mutated in various human tumors. Polymorphism is an additional genetic alteration observed in exons and introns of the p53 gene of normal tissues and tumors. Distributions of alleles of three common polymorphisms of the p53 gene; a 16 bp duplication in intron 3, codon 72 of exon 4 and a sequence in intron 6, were studied in peripheral white blood cells (WBC) of patients with ovarian or endometrial carcinomas. The analysis was performed by PCR and direct sequencing. The 100% linkage observed between the most common haplotypes of each polymorphism in healthy subjects was lower in the patients. A significant difference was observed between frequencies of genotype and haplotype combinations in patients with ovarian carcinoma and endometrial carcinoma. The incidence of heterozygosity was increased in ovarian carcinoma and decreased in endometrial carcinoma. Our results suggest that the p53 gene may be involved in susceptibility and predisposition to various cancers not only by mutations but also by preferential presentation of polymorphic alleles. PMID- 9864428 TI - Cytological diagnosis and telomerase activity of cells in effusions of body cavities. AB - Telomerase is a ribonucleoprotein that synthesizes telometric DNA on chromosome ends, and may be related to the aging and immortality of cells. Recently, a telometric repeat amplification protocol (TRAP) assay for telomerase activity, using the polymerase chain reaction, was developed. We examined the limitations of TRAP assay by applying it to a cultured colon cancer cell line (COLO320) and 58 human cytological materials from body cavity effusions, and obtained the following results; i) The limits of the TRAP assay were 20-50 cells for the COLO320 cell line; ii) One COLO320 cell per 100 normal blood white cells was detectable; iii) Seventeen of 58 samples were positive for telomerase activity in this study. The sensitivity was 69% (9/13) and the specificity was 87.5% (28/32) between cytological diagnosis and telomerase activity; iv) Among 29 malignant cases, 15 were positive for telomerase activity, while there were 11 cytologically positive cases. The positive cases detected by the combination of cytology and telomerase activity accounted for 21 of the total 29 cases (72.4%). These results suggest that the measurement of telomerase activity in body cavity effusions may be useful as an adjunctive tool for cytological and clinicopathological diagnosis and that this technique is potentially applicable to remnant cytological materials. PMID- 9864429 TI - Absence of epidermal growth factor receptor expression in squamous cell carcinoma of the uterine cervix is an indicator of limited tumor disease. AB - The expression of growth factors is considered as an important diagnostic and prognostic feature in tumor pathology. We investigated the value of the immunohistochemical EGF-receptor expression (EGF-R) in 30 squamous cell carcinomas of the uterine cervix, treated by radical hysterectomy and lymphadenectomy according to the Wertheim-Meigs-Okabayashi technique. Immunohistochemical reactions were performed on 4 microm sections from paraffin embedded tissue, using an indirect peroxidase method. The staining results were evaluated semiquantitatively as negative (n=9; 30%) or as slightly, moderately or severely positive (n=21; 70%). The EGF-R-negative tumors were found in less advanced tumor stages. None had invaded into the parametrium (100%), eight were staged as T1 (89%), seven as N0 (78%), and seven showed no evidence for lymphangiosis carcinomatosa (78%). The respective values for the EGF-R-positive tumors ranged from 52% to 67%. However, only the difference in parametral invasion (EGF-R-negative: 0%, EGF-R-positive: 38%) was statistically significant (p=0.0306), probably due to the small number of cases. The EGF-R-expression was not correlated to histomorphological tumor grading. The results of this study indicate an inverse correlation between EGF-R expression and tumor spread. Assuming that this trend could be confirmed by a larger group of patients, immunostaining for EGF-R in a tumor biopsy could be useful to adapt surgical strategies and adjuvant therapy in the individual patient. Moreover, the EGF-R is an interesting target for immunotherapeutic approaches in squamous cell cervical carcinoma. PMID- 9864430 TI - DNA vaccine against oncogenic hamster cells transformed by HPV16 E6/E7 oncogenes and the activated ras oncogene. AB - The capability of DNA to elicit anti-tumour immunity was studied using human papillomavirus type 16 (HPV16)-transformed Syrian hamster cells denoted K3/II. These cells had been derived after cotransfection of primary kidney cell cultures with p16HHMo plasmid containing E6/E7 oncogenes of HPV16 and pEJ6.6 plasmid containing the activated human H-ras oncogene; they express both the HPV16 and activated H-ras genes. As a DNA vaccine, the p16HHMo plasmid was used. Three doses of the plasmid (either 100 microg or 10-15 microg per dose) were administered intramuscularly at 3-week intervals. The animals were challenged with four different doses (10(3)-10(6) per animal) of K3/II cells 10 days after the last plasmid injection. In one experiment the lower dose of plasmid DNA was also given in a mixture with the cationic lipid DOTAP. In another experiment, the pEJ6.6 plasmid (100 microg per dose) was used either alone or in combination with p16HHMo. In all experiments animals inoculated with the same doses of pBR322 plasmid served as controls. A moderate protective effect was observed in animals inoculated with the 100-microg doses of p16HHMo, but not in those inoculated with 10-15 microg of the same plasmid, whether given with or without DOTAP. A protective effect was also observed after administration of the pEJ6. 6 plasmid. At the time of challenge a portion of the p16HHMo-immunized, but not the pBR322 treated, animals possessed antibodies reactive in ELISA with peptides derived from the N-terminal portion of HPV16 E7 protein and with one peptide derived from E6 protein, while two other E6 peptides exhibited non-specific reactivity. PMID- 9864432 TI - Alternative exon-specific PCR method for the analysis of human CD44 isoform expression. AB - CD44 is a widely expressed cell-surface transmembrane glycoprotein involved in diverse adhesive processes. Its isoforms have been implicated in tumor progression and are considered a promising marker for evaluation of the metastatic potential of various tumors. Several methods have been described for the analysis of CD44 isoforms in tumor cells, including immuno-histochemistry, RT PCR followed by hybridization and nested RT-PCR. We describe an alternative nested PCR for the analysis of CD44 isoform expression in various malignancies. Total RNA was isolated from various shock-frozen tissues from human tumors, reverse-transcribed and PCR-amplified using CD44-specific primers. Reverse transcription was performed by two different methods, either using Tth-polymerase or MMLV-RT. Exon-specific amplification was then carried out using specific primers for each variable exon. Amplification products were assayed by agarose gel electrophoresis. Comparison of the patterns obtained from the first amplification and from the exon-specific amplification allowed to identify exons expressed by tumor tissues, as well as the genomic organization of CD44 isoforms. The method developed proved to be sensitive, reliable and inexpensive in comparison with other methods. It can be performed even in solid tumors and for numerous samples, and is suitable for laboratories with limited resources. PMID- 9864431 TI - The chemosensitizing potential of GF120918 is independent of the magnitude of P glycoprotein-mediated resistance to conventional chemotherapeutic agents in a small cell lung cancer line. AB - GF120918, at 250 ng/ml, increased the sensitivity of a P-glycoprotein (P-gp) mediated multidrug resistant (MDR) small cell lung cancer cell line (H69/LX4) to the P-gp substrates, paclitaxel, taxotere, vinblastine, vinorelbine, daunorubicin and etoposide to levels which were either greater (in the case of etoposide) or close to that of the parent cell line (H69/P). This was achieved in spite of the great variation in the levels of resistance of the MDR cell line for the various anti-cancer drugs tested. These data suggest that GF120918 is a potent antagonist of P-gp mediated multidrug resistance, even in the case of high levels of resistance, as was the case with paclitaxel and taxotere (2560 and 2215 fold more than the sensitive parent cell line respectively). PMID- 9864433 TI - Effect induced by interleukin-1 on the behaviour of B16F10 melanoma cells. AB - We have previously demonstrated that in vitro treatment with interleukins (IL-2 and IL-6) enhances metastasis in B16 murine melanoma. IL-1 has been reported to be involved in the metastatic activity of human melanoma; so in the present study we have investigated if IL-1 could also modulate the behaviour of this experimental tumor. Data showed that low doses of IL-1 induced cell proliferation and several modifications in the expression of MHC antigens, fibronectin and laminin, however these changes did not lead to a higher metastatic efficiency. In addition it was shown that B16F10 cells express IL-1 mRNA. It can be concluded that IL-1 may be involved in the biology of B16F10 cells although its role in metastasis needs further investigation. PMID- 9864434 TI - Analysis of the cyclin-dependent kinase inhibitor p27Kip1 in muscle invasive bladder cancer. AB - It has been suggested that a deregulated cell cycle control contributes to the development of human malignancies due to the loss of critical antiproliferative mechanisms. The cell cycle is controlled at two checkpoints, one at the G1-S and another at the G2-M transition. Several genes including the structurally related p21WAF/CIP1 gene, the downstream mediator of the p53 tumor suppressor gene, and the p27Kip1 gene have been identified as inducers of cell cycle arrest at the G1 checkpoint when substantial DNA damage has occurred to avoid further replication of the altered genome. Recently, a heat stable 27 kDa protein, the transcript of the p27Kip1 gene, has been identified and was suggested to substantially participate in cell cycle control at the G1 checkpoint. Previous investigations have correlated decreased expression of the p27Kip1 protein with an increased biological aggressiveness of breast and small cell lung cancer. However, the molecular-genetic analysis of a variety of human malignancies including prostate cancer failed to identify any alteration at the p27Kip1 gene locus, therefore suggesting a loss of p27Kip1 protein expression to result from post transcriptional/post-translational events or from so far unknown regulatory mechanisms. So far, bladder cancer specimens have neither been investigated for p27Kip1 alterations on the DNA level, nor has the result of molecular genetic analysis been correlated with an immunohistochemically detected expression of the gene product, the p27Kip1 protein. The present study is the first to describe p27Kip1 gene alterations on the DNA level in 3 of 42 muscle invasive bladder cancer specimens. In contrast, loss of p27Kip1 protein expression was observed in 14 of 42 (33%) tumors. According to the previously reported observation in a variety of human malignancies, in bladder cancer loss of p27Kip1 protein expression seems to result from post-transcriptional or post-translational events. PMID- 9864436 TI - Von Neumann's quintessential message: genotype + ribotype = phenotype. AB - In this short article, we argue that von Neumann's quintessential message with respect to self-replicating automata is genotype + ribotype = phenotype. Self replication occurs in analogy to nature: The description (genotype) written on the input tape is translated via a ribosome (ribotype) so as to create the offspring universal constructor (phenotype). PMID- 9864435 TI - Relationships betxween Fas expression, activation molecule CD25, and functional activity of tumor-associated lymphomonocytes from neoplastic effusions. AB - Fas ligand (FasL), a cell surface molecule belonging to the tumor necrosis factor family, binds to its receptor Fas, thus inducing apoptosis of Fas bearing cells. In the present study we assessed the expression of Fas, activation molecule interleukin (IL)-2 receptor alpha chain (CD25) and an index of functional activity such as thymidine uptake under mitogen stimulation of tumor associated lymphomonocytes (TALM) from 7 neoplastic effusions of advanced cancer patients. The same parameters were studied in peripheral blood mononuclear cells (PBMC) of 7 patients with cancer of different sites and in 7 normal subjects. The proliferative response to phytohemagglutinin (PHA), measured as [3H]-thymidine uptake, of TALM was significantly lower than that of PBMC of cancer patients. The expression of CD25 on unstimulated fresh TALM was slightly higher than that of PBMC from normal subjects: after 24 h of PHA stimulation the CD25 was expressed both on TALM and PBMC from normal subjects. The expression of Fas was assessed on unstimulated TALM, PBMC from cancer patients and normal subjects immediately after (by 2 h, t0) the cell separation, and at different times (24 h and 48 h) thereafter, and on PHA-stimulated TALM, PBMC from cancer patients and normal subjects after 24 h and 48 h of culture (in RPMI 1640). At all times (t0, 24 h and 48 h) the Fas expression by unstimulated TALM was significantly higher than that of PBMC from normal subjects: the Fas expression by PBMC from cancer patients was roughly in the same range as PBMC from normal subjects. At 24 h the Fas expression by PHA-stimulated TALM was significantly higher than that of PBMC from normal subjects, whereas at 48 h the difference was not significant. The TALM studied by us showed to be functionally defective and expressing relatively high levels of Fas showing the characteristics to be considered as a target for FasL expressing tumor cells, which in this way may escape immune control. PMID- 9864437 TI - John von Neumann: the founding father of artificial life. AB - Aside from being known for his contributions to mathematics and physics, John von Neumann is considered one of the founding fathers of computer science and engineering. Not only did he do pioneering work on sequential computing systems, but he also carried out a major investigation of parallel architectures, leading to his work on cellular automata. His exceptional vision and daring, borrowing from biology the concept of genomic information even before the discovery of DNA's double helix, led him to propose the concept of self-reproducing automata. PMID- 9864438 TI - Fifty years of research on self-replication: an overview. AB - The study of self-replicating structures or machines has been taking place now for almost half a century. My goal in this article is to present an overview of research carried out in the domain of self-replication over the past 50 years, starting from von Neumann's work in the late 1940s and continuing to the most recent research efforts. I shall concentrate on computational models, that is, ones that have been studied from a computer science point of view, be it theoretical or experimental. The systems are divided into four major classes, according to the model on which they are based: cellular automata, computer programs, strings (or strands), or an altogether different approach. With the advent of new materials, such as synthetic molecules and nanomachines, it is quite possible that we shall see this somewhat theoretical domain of study producing practical, real-world applications. PMID- 9864439 TI - Self-replicating and self-repairing multicellular automata. AB - Biological organisms are among the most intricate structures known to man, exhibiting highly complex behavior through the massively parallel cooperation of numerous relatively simple elements, the cells. As the development of computing systems approaches levels of complexity such that their synthesis begins to push the limits of human intelligence, engineers are starting to seek inspiration in nature for the design of computing systems, both at the software and at hardware levels. We present one such endeavor, notably an attempt to draw inspiration from biology in the design of a novel digital circuit: a field-programmable gate array (FPGA). This reconfigurable logic circuit will be endowed with two features motivated and guided by the behavior of biological systems: self-replication and self-repair. PMID- 9864440 TI - Self-replicating structures: evolution, emergence and computation. AB - Since von Neumann's seminal work around 1950, computer scientists and others have studied the algorithms needed to support self-replicating systems. Much of this work has focused on abstract logical machines (automata) embedded in two dimensional cellular spaces. This research was motivated by a desire to understand the basic information-processing principles underlying self replication, the potential long-term applications of programmable self replicating machines, and the possibility of gaining insight into biological replication and the origins of life. We view past research as taking three main directions: early complex universal computer-constructors modeled after Turing machines, qualitatively simpler self-replicating loops, and efforts to view self replication as an emergent phenomenon. We discuss our recent studies in the latter category showing that self-replicating structures can emerge from nonreplicating components, and that genetic algorithms can be applied to program automatically simple but arbitrary structures to replicate. We also describe recent work in which self-replicating structures are successfully programmed to do useful problem solving as they replicate. We conclude by identifying some implications and important research directions for the future. PMID- 9864442 TI - Effect of hydrophobization of basic pancreatic proteinase inhibitor on the inhibition of bovine trypsin and human leukocyte elastase. AB - The effect of modification of basic pancreatic trypsin inhibitor (BPTI) by derivatives of fatty acids (oleic, stearic) on the inhibition of bovine trypsin and human leukocyte elastase (HLE) was studied. Kinetic constants of interaction with trypsin and inhibition constants of both enzymes were determined. Hydrophobization of BPTI had virtually no effect on its high affinity for trypsin. The coupling of cis-unsaturated oleoyl radicals to the inhibitor molecule significantly increased the efficiency of HLE inhibition, whereas the coupling of saturated stearoyl radicals completely canceled the anti-elastase activity and in some cases promoted the substrate hydrolysis. PMID- 9864443 TI - Detection and partial characterization of bacteriocin in the methanotrophic bacterium Methylobacter bovis. AB - The strain Methylobacter bovis 98 was selected among methanotrophic bacteria as one of the most active producers of secretory bacteriocin-like compounds. In the above strain this compound was shown to be a protein with a molecular weight of about 70 kD, relatively thermostable, having a bactericidal effect on closely related organisms. Its properties as a whole are consistent with the accepted definition of bacteriocins, which so far have not been found in this group of microorganisms. A methodical approach that combines electrophoretic separation of secretory proteins and testing their antibacterial activity directly in polyacrylamide gel allowed us for the first time to identify bacteriocin in methanotrophic bacterial culture. PMID- 9864444 TI - Structural organization and phase behavior of DNA-calcium dipalmitoylphosphatidylcholine complex. AB - Freeze-fracture study of ultrastructure of DNA--calcium- dipalmitoylphosphatidylcholine (DPPC) complex was carried out at different temperatures. For high-speed cryofixation from controllable initial temperatures, a special thermostatic chamber was designed. The fracture surface of the complex was found to be considerably different from the initial DPPC liposomes: 1) the period of ripple phase was 25 nm in contrast to 15 nm for control samples; 2) the ripple phase was observed at temperatures ranging from 6 degreesC to lipid melting temperature; 3) at temperature above the lipid melting unordered worm like folds were formed on the fracture surface. Their length was correlated with the length of DNA fragments used in the experiment. We suppose that DNA molecules adsorbed on the membrane surface were segregated to clusters, resulting in formation of a new phase with specific structure and properties. PMID- 9864447 TI - Energetic aspects of CO oxidation in desulfovibrio desulfuricans AB - In cell suspension of Desulfovibrio desulfuricans B-1388, oxidation of CO as the only energy source is associated with reduction of SO42-. After a 2-h incubation of cells in 8% CO, 81% of the gas is converted. Oxidation of 1 mole CO results in formation of 0.23 mole H2S. Intracellular ATP content increases from 2.5 (control) to 8.3 nmoles/mg (during CO conversion). Dinitrophenol inhibits sulfate reduction and CO oxidation. CO dehydrogenase was detected in cytoplasmic and membrane cell fractions (59 and 34%, respectively). PMID- 9864445 TI - Analysis of antibodies that recognize the B-epitopes formed on protein antigens by glutaraldehyde treatment: an efficient method of their neutralization. AB - Immunochemical analysis of antigenic determinants (AD) formed on protein macromolecules by glutaraldehyde (GA) treatment with subsequent block of free aldehyde groups with glycine was performed. The structure of the epitopes was analyzed using BSA and ovalbumin (Ova) treated with GA with subsequent block of the active groups with glycine and other amino acids. The products of reaction of GA with Gly, Lys, Pro, and His were used as hapten-like antigens that mimic GA antigenic determinants (GA-AD). Indirect and competitive enzyme-linked immunoassay and immunoaffinity chromatography revealed two regions of epitope density in the structure of the GA determinant. It is shown that the product of GA reaction with Gly efficiently blocks the antibodies raised against GA-treated proteins. Based on this finding, a method of assay of the peptide-specific antibodies of the immune complexes with the peptide--protein conjugate in the presence of antibodies to the linking agents is suggested. PMID- 9864446 TI - Vitamin E and A deficiencies in children correlate with Chernobyl radiation loads of their mothers. AB - Essential lipid bioantioxidants (BAO) (vitamins E and A) and catabolic products of the lipoperoxide cascade (LPC) (diene conjugates, ketodienes, and carbonyls) were determined in blood plasma of children aged 11 days to three years (n = 123) who are residents of regions contaminated with radionuclides (5-40 Ci/km2 of 137Cs) discharged during the Chernobyl power plant accident. Individual antenatal radiation loads Da of children and total radiation doses of their mothers Dm were also determined as external gamma-radiation loads, which correlate with internal radiation doses for permanent residents of contaminated regions. Before examination, therapy with multivitamins containing BAO determined in this work was administered to groups of pregnant women and their children (group I, n = 48) age-matched to a group that received no therapy (group II, n = 99). In group II, reduced concentrations of vitamins E and A decreased to levels indicating severe vitamin deficiency and displayed inverse correlations with Dm and Da. However, in group I these decreases and inverse correlation were only characteristic of initially normal levels of tocopherol. In group II, increases in Dm and Da caused stress-related increases in blood levels of diene conjugates and ketodienes (which was stronger than in group I with increases in Da) and, because of this, E and A vitamin deficiencies. In group I, LPC levels inversely correlated with Dm. We found nine significant correlations of radioloads of mothers with BAO and LPC pathologies of their children. The total pathogenic effectiveness of Dm in relation to LPC and BAO of children is comparable to the effectiveness of Da, which is the most hazardous radiation load. The possibility of existence of a radiation-induced protective response mobilizing BAO from depots into the blood stream is considered. BAO therapy increased the radiation resistance of children. A hypothesis was formulated that the uptake by children of their mother's radioactive Chernobyl isotopes in amounts adequate to their radiation loads is the mechanism of correlations of Dm with long-term pathogenic effects on BAO and LPC systems of children. PMID- 9864448 TI - Interaction of steroid--peroxidase conjugates with cellulose immunosorbents in aqueous and micellar media. AB - In 0.1 M bicarbonate buffer (pH 9.0) and in microemulsions of aerosol OT (AOT) and its mixture with Triton X-45 in heptane, antibodies against cortisol (anti COR) and progesterone (anti-PROG) were covalently immobilized on fine-porous cellulose filters (0.6 cm diameter) after sodium periodate oxidation. Immunosorbents obtained in different media were characterized in terms of antibody-bound density and antigen-binding capacity with respect to peroxidase- steroid conjugates HP--COR-11 and HP--PROG-4 containing 11 molecules of cortisol and four progesterone molecules, respectively. For all immunosorbents antigen binding capacity expressed as peroxidase activity of immune complexes formed on the solid cellulose phase in aqueous and micellar media was determined. Dissociation constants of immunocomplexes on the cellulose formed in aqueous and micellar media were determined using ELISA. In all cases Kd values in aqueous media were approximately 10-8 M and were significantly lower than corresponding values of dissociation constants of immune complexes in mixed microemulsions of AOT and Triton X-45. PMID- 9864449 TI - Spectrophotometric quantitative analysis of the main hemoglobin derivatives. AB - A spectrophotometric multiwavelength method for the simultaneous determination of the main hemoglobin derivatives including oxyhemoglobin, deoxyhemoglobin, carboxyhemoglobin, and methemoglobin has been developed. The choice of analytical wavelengths was made by the methods of information factors and linear programming. The software for the quantitative analysis was developed considering the least square method, linear programming, algebraic background correction, and combined methods including linear programming and algebraic background correction. PMID- 9864450 TI - Binding of apoB-containing lipoproteins from unfractionated human blood sera to immobilized LDL receptor. AB - Binding of apoB-containing lipoproteins from unfractionated human blood sera to the immobilized bovine receptor of low density lipoproteins (LDL receptor) was studied. Peroxidase-labeled anti-human apoB antibodies were used to evaluate the lipoprotein binding. The equilibrium dissociation constant (Kd) of the interaction between apoB-containing lipoproteins from unfractionated human sera from healthy donors and the immobilized LDL receptor varied from 1 to 20 microg apoB/ml. To describe the binding of lipoproteins to the LDL receptor, a parameter of relative binding affinity (RBA) was used. RBA is inversely related to value of Kd and equal to unity for the standard serum. The RBA values for the binding of apoB-containing lipoproteins from unfractionated sera to the immobilized LDL receptor were found to correlate with the RBA values for the binding of isolated VLDL (r = 0.76, p < 0.001) and fail to correlate with the RBA values for the binding of isolated LDL. The RBA values for the binding of apoB-containing lipoproteins from unfractionated sera correlated with the RBA values for the binding of apoE-containing lipoproteins from unfractionated sera (r = 0.92, p < 0.001) and with values of triglyceride concentration in the sera (r = 0.93, p < 0.001). The RBA values for the binding of apoB-containing lipoproteins from sera of patients with FDB whose LDL were unable to bind to the LDL receptor did not significantly differ from the RBA values for the normal sera. However, the removal of VLDL from the normal sera significantly decreased the RBA values for the binding of apoB-containing lipoproteins from unfractionated sera. The results indicate that the different binding of apoB-containing lipoproteins to the immobilized LDL receptor mainly depended on the different binding of VLDL and not of LDL. PMID- 9864451 TI - Study of intracellular localization and traffic of newly synthesized ceruloplasmin receptor in cultured human fibroblasts. AB - Ceruloplasmin (Cp) receptor in cells of non-hepatocyte lineage (human HT-1080 fibroblasts) is synthesized by membrane-bound polyribosomes and then becomes a resident of the plasma membrane. The intracellular traffic of [14C]Cp receptor was followed in pulse-chase experiments using specific antibodies. It was shown that pulse-labeled Cp receptor, after reaching the place of its residence in the plasma membrane, is retained there for 90 min and then appears in the cytosol. Immunoactive 20-kD fragments of Cp receptor were found in the culture medium 1.5 h later. The intracellular traffic of 125I-labeled Cp bound to the fibroblast cell surface was traced in parallel chase experiments. It was shown that the internalized Cp receptor was recovered from the floating fraction of the cytosol. Comparison of the dynamics of the retention of internalized [14C]Cp receptor and 125I-labeled Cp in the subcellular compartments demonstrated that the traffic of both proteins within the fibroblasts is coordinated in time and proceeds via a common route. The role of Cp receptor in copper uptake by non-hepatocyte cells is discussed. PMID- 9864452 TI - Isolation and properties of extracellular alkaline phosphatase from Bacillus intermedius. AB - Alkaline phosphatase (APase) was isolated from the culture liquid of the streptomycin-resistant strain of Bacillus intermedius S3-19 and purified as a homogeneous preparation by ion-exchange chromatography and FPLC. Electrophoresis and gel-filtration revealed that the active enzyme is a monomer with molecular weight of 46-47 kD. The enzyme possessed phosphomonoesterase and phosphodiesterase activities with maximal levels at pH 9.5 and 55 degreesC and was stable until 60 degreesC at pH 8.0-10.0. The isolated APase exhibits a broad specificity towards a wide variety of substrates. The effect of divalent metal ions and other reagents on its catalytic activities was studied. It was concluded that alkaline phosphatase of B. intermedius is similar to the secreted alkaline phosphatases from other Bacillus species in its physicochemical and catalytic properties. PMID- 9864453 TI - Tryptophan residues in alpha-galactosidase from Trichoderma reesei. AB - Tryptophan residues in alpha-galactosidase were modified with bromosuccinimide. The fact that galactose, a specific inhibitor of alpha-galactosidase, does not prevent this modification demonstrates that tryptophan residues are not located in galactose binding sites. Analysis of the inactivation kinetics revealed two groups of Trp residues (8.5 and 7.5 residues) with different accessibility for N bromosuccinimide. We studied specific quenching of alpha-galactosidase fluorescence resulting from modification of an sulfhydryl group in the active site of the enzyme with Hg2+ and Ag+ ions. The specific quenching is due to conformational changes of the enzyme. Forster's radii were determined for various protein--chromophore complexes. Dynamic quenching of alpha-galactosidase fluorescence was investigated. To describe abnormal dynamic quenching in alpha galactosidase, a modification of the Stern--Volmer equation is suggested. PMID- 9864455 TI - Structure of sulfated O-specific polysaccharide of the marine bacterium Pseudoalteromonas marinoglutinosa KMM 232. AB - A sulfated O-specific polysaccharide containing D-mannose, L-rhamnose, and the sulfate group was obtained by mild acid hydrolysis of lipopolysaccharide (S-form) of the marine bacterium Pseudoalteromonas marinoglutinosa KMM 232. Based on analysis of methylation and 13C-NMR spectroscopy of native and desulfated polysaccharides, the following structure of disaccharide repeat unit in the O specific polysaccharide has been established: [scheme]. This is the first report of a sulfated O-specific polysaccharide isolated from gram-negative bacteria. PMID- 9864454 TI - Peroxidase activity of ferritin in aerosol OT reversed micelles in heptane. AB - Using oxidation of o-phenylenediamine (PDA) and tetramethylbenzidine (TMB) by hydrogen peroxide, cumene peroxide (CHP), tert-butyl hydroperoxide (TBHP), and Triton X-45 hydroperoxide (Triton X-45-HP), the peroxidase activity of horse spleen ferritin was investigated in reversed micelles of aerosol OT (AOT) in heptane with various hydration degrees. With hydrogen peroxide as oxidant the dependences of initial rate of oxidation of both substrates (v0) on hydration degree W0 are characterized by maxima at W0 = 9-11, 20, and 41. In the system containing TBHP--ferritin these maxima were not observed. The parameters kcat, Km, and their ratios kcat/Km as a criterion of ferritin efficiency in peroxidase reactions were determined for both substrates in micellar medium at various W0. Increase of W0 was accompanied by a decrease of kcat and Km. With hydrogen peroxide the peroxidase activity of ferritin in the AOT micelles was significantly lower than in 0.1 M acetate buffer, pH 4.2. However, the efficiency (expressed as kcat/Km) of a system ferritin--Triton X-45-HP in micellar TMB oxidation exceeded that in the aqueous medium. A method of purification of iron containing crystallite from the ferritin molecule was developed using reversed AOT micelles in heptane and heating the mixture on a water bath. PMID- 9864456 TI - Effect of modification of histidine residues on organization of the pigment- protein complexes of chromatium minutissimum AB - The effect of diethyl pyrocarbonate on chromatophores and isolated pigment- protein complexes of Chromatium minutissimum was studied. It is shown that modification of histidine residues results in the destruction of the core antenna LHI (B880) and in a spectral shift from 850 to 830 nm in the peripheral antenna LHII (B800-850). In the purple sulfur bacterium Chromatium minutissimum the pigment--protein complexes B800-B850 (peripheral antenna, LHII) and B880 (core antenna, LHI) collect and transmit the absorbed light energy to the reaction centers. The composition of pigments and proteins as well as primary structure of the majority of polypeptides in both types of complexes from various photosynthetic bacteria have been determined. PMID- 9864457 TI - Alpha-galactosidase of the marine bacterium Pseudoalteromonas sp. KMM 701. AB - An alpha-galactosidase that inactivates the group specificity of B erythrocytes (group III) of human blood and does not affect A erythrocytes (group II) was isolated from the marine bacterium Pseudoalteromonas sp. KMM 701. The enzyme preparation did not contain lectin, hemolytic, sialidase, endoglycanase, or glycosidase activities. The enzyme is stable at 20 degreesC for 24 h, has pH optimum for catalysis within the range of 6.7-7.7, and is stable to high concentrations of NaCl. It is 4-fold more efficient than the alpha-galactosidase from green coffee beans. At pH 7.0 the Km for p-nitrophenyl-alpha-D galactopyranoside is 0.29 mM. The molecular weight of the enzyme determined by gel-filtration is 195 +/- 5 kD. The alpha-galactosidase is denatured by urea and guanidine hydrochloride. Its activity does not depend on the presence of metal ions. It contains a sulfhydryl group essential for its catalytic activity. PMID- 9864458 TI - Increased functional activity of elongation factor G with G16V mutation in the GTP-binding domain. AB - Oligonucleotide-directed mutagenesis was used to obtain elongation factor G from Thermus thermophilus with the G16V mutation in its GTP-binding domain. Functional studies of the mutated protein and elongation factor G from E. coli were carried out. The data revealed that the G16V mutant retains high thermostability, has an increased ribosome-dependent GTPase activity, and its translation activity in cell-free translation system is equal to that of the factor G from E. coli. The mutated protein with an uncleavable GTP analog also has an increased affinity to the ribosomes. PMID- 9864459 TI - Study on splitting of sucrose glycoside bonds by beta-fructosidase of Aspergillus niger VKM F-801. AB - Functional groups of beta-fructosidase of Aspergillus niger VKM F-801 were identified. Based on the pH dependence of the enzyme, the calculated heat of ionization, the photoinactivation of the enzyme in the presence of methylene blue photosensitizer, and also the enzyme inactivation by diethyl pyrocarbonate, the catalytic center of the beta-fructosidase was found to include carboxyl and imidazole groups of amino acid residues. The glycoside bond in the sucrose molecule is apparently cleaved by the catalytically active carboxylate--imidazole pair. PMID- 9864460 TI - Phospholipid pool, lipid peroxidation, and superoxide dismutase activity under various types of oxidative stress of the brain and the effect of low-energy infrared laser irradiation. AB - The effect of low-energy infrared laser irradiation on the phospholipid pool, lipid peroxidation, and superoxide dismutase activity in the brain of white rats was studied in experimental ischemia, reperfusion, and acute edema. These models are characterized by oxidative stress; the contents of tri- and diphosphoinositides and sphingomyelins were lowered, whereas the levels of phosphatidylserine and phosphatidylethanolamine did not change, and the amount of phosphatidylcholine was increased. In acute brain edema, the contents of hydroperoxides and malonic dialdehyde in enzymatic and nonenzymic lipid peroxidation systems were increased in mitochondrial and microsomal fractions and the level of arachidonic acid was significantly elevated. Infrared laser irradiation contributes to the correction of the changes in the phospholipid pool; laser irradiation lowered the increased levels of hydroperoxides and malonic dialdehyde and elevated superoxide dismutase activity in the brain during ischemia, reperfusion, and acute edema of the brain. The data suggest that low energy infrared laser irradiation has certain neuroprotective activity in various types of oxidative stress including ischemia, reperfusion, and acute edema of the brain. PMID- 9864461 TI - Polyclonal antibodies from blood and cerebrospinal fluid of patients with multiple sclerosis effectively hydrolyze DNA and RNA. AB - It is known that in the blood of patients with some autoimmune diseases catalytically active antibodies hydrolyzing proteins, DNA, and RNA may be detected. In the present work homogeneous preparations of IgG antibodies (Ab) possessing high affinity for nucleic acids (NA) were obtained for the first time from blood and cerebrospinal fluid of patients with multiple sclerosis (MS). The fraction of IgG Ab as well as its Fab fragments and isolated light chains of both kappa- and lambda-types were shown to catalyze effectively the hydrolysis of DNA and RNA. It is shown by different methods that the capability for nucleic acid hydrolysis is an intrinsic property of the polyclonal Ab. NA-hydrolyzing Ab were detected in the blood of 69 of 72 and in the cerebrospinal fluid of 5 of 5 examined MS patients, while they were not detected in the blood of any of 50 healthy donors examined. Comparison of relative rates of RNA hydrolysis and of the substrate specificity in hydrolysis of various model RNAs--cCMP, poly(U), poly(A), and poly(C)--revealed pronounced differences of MS antibodies from ribonucleases of human blood, ribonuclease A, and all earlier described abzymes. The abzymes are usually characterized by relatively low specific activities in comparison with that of normal enzymes catalyzing analogous reactions. Ab from the blood of MS patients are the first example of autoabzymes whose specific activity in RNA hydrolysis is comparable or even higher than that of pancreatic ribonuclease A--one of the most active RNA-hydrolyzing enzymes. PMID- 9864462 TI - Molecular mechanisms of diabetic complications. AB - The main molecular process responsible for the development of diabetic complications is the activation of intracellular protein kinase C by long-term hyperglycemia. In ordinary somatic cells (e.g., endothelial or retinal cells) this activation disturbs the metabolism of these cells, whereas a transition into an "excited state" of effector cells such as monocytes or polymorphonuclear leukocytes causes an avalanche-like increase of various pathological processes damaging the body. In relatively compensated diabetes mellitus, when the duration of hyperglycemia is not long and levels of hyperglycemia are not too high, the intensity of the above-mentioned processes is low. However, due to the long time course of reparative processes, some damages which appear in short periods of decompensation can accumulate in the body, finally causing the clinical manifestation of vascular complications of diabetes mellitus. PMID- 9864463 TI - New methods of protein purification. Displacement chromatography. AB - This review discusses a new method for protein purification, displacement chromatography. Elution of proteins bound to an adsorbent is carried out in displacement chromatography by a substance, the so-called "displacer", which has a higher affinity for the chromatographic matrix than any of the adsorbed proteins. The latter are "pushed out" from the matrix by the displacer and form a displacement train when moving along the column. The component with the weakest affinity moves first, and the one with the highest affinity moves last in the train. The concentration of the component in the displacement zone is determined by the intersection of the operating line and binding isotherm of the given component. The shape of the displacement zone is close to rectangular. A more powerful resolving force is created compared to other methods of selective elution, especially at high column loadings, when nearly the whole working capacity of the column is used. Independence of component concentration in the displacement zone on its content in the feed allows significant concentrating of the purified protein during displacement chromatography. Examples of application of displacement chromatography for the separation of model protein systems and purification of proteins from crude extracts are discussed as well as methods of column regeneration. PMID- 9864464 TI - Thermostable DNA-polymerase from Thermus thermophilus B35: isolation and characterization of some properties. AB - Thermostable Tte DNA-polymerase was isolated from the strain Thermus thermophilus B35 which was found in hot spring water. The enzyme with molecular mass 87 kD was isolated using sequential chromatography on DEAE-Sepharose, hydroxylapatite, hexyl-agarose, and heparin-Sepharose. Biochemical properties of Tte DNA polymerase are similar to those of Tth DNA-polymerase isolated from Thermus thermophilus HB8; however, practical application of Tte-Pol seems to be more favorable due to higher temperature optimum of this enzyme and lack of restriction endonucleases in the initial strain. PMID- 9864465 TI - Proteoglycan composition in cell nuclei of mouse hepatoma. AB - Cell nuclei of mouse hepatoma contain various proteoglycans (PG) which include heparan sulfate proteoglycan (HS-PG), dermatan sulfate proteoglycan (DS-PG), and chondroitin sulfate proteoglycan (CS AC-PG). The latter is not found in cell nuclei of normal mouse liver. Heparan sulfate (HS) and dermatan sulfate (DS) are the main constituents of carbohydrate chains of nuclear proteoglycans of tumor and normal cells, respectively. Changes in the composition of nuclear PG during malignant transformation are discussed considering the concept of their possible involvement in the regulation of cell mitotic activity. PMID- 9864466 TI - Effects of medium viscosity on kinetics of the enzymatic reaction catalyzed by bacterial RNase AB - Effects of medium viscosity on kinetic parameters of poly(U) hydrolysis catalyzed by RNase from Bac. intermedius 7P (binase) were studied in solutions of sucrose (4-50 wt. %) and glycerol (35-62 wt. %) in Tris--sodium acetate buffer (pH 7.5) at 25 degreesC. The rate constant of reaction kcat was practically unchanged over a wide range of viscosities (1-15 cP for sucrose and 2.5-3 cP for glycerol). In glycerol solutions, kcat slightly increased with viscosity increase from 4 to 10 cP. Addition of NaCl to the buffer medium resulted in an inhibitory effect of Na+ on kcat, prevented by 50% sucrose or 60% glycerol. It is concluded that binase catalyzed poly(U) cleavage occurs through a "tense"-substrate mechanism, similarly to reactions catalyzed by alpha-chymotrypsin, trypsin, and laccase. PMID- 9864467 TI - Isolation of rabbit liver heat shock protein with molecular weight 90 kD (Hsp90) and its interaction with troponin components and calponin. AB - Using a modified method consisting of chromatography on phenyl-Sepharose, Q Sepharose, and hydroxyapatite, we isolated a highly purified heat shock protein with molecular weight 90 kD (Hsp90) from rabbit liver. The isolated protein was recognized on immunoblot by commercially available monoclonal anti-Hsp90 antibodies. The chromatographic properties, interaction with actin and calmodulin, phosphorylation in the presence of Mg-ATP, and one-dimensional peptide maps of rabbit liver Hsp90 are similar to the corresponding properties of Hsp90 isolated from other sources. In the presence of soluble carbodiimide and N hydroxysuccinimide, rabbit liver Hsp90 can be cross-linked with calmodulin, troponin C, troponin I, and calponin. The data obtained indicate that Hsp90 may participate in the assembly of regulatory proteins of the actin filament. PMID- 9864468 TI - Thermodynamic and stereochemical parameters to evaluate stability of hydrophobic cores of globular proteins. AB - This work presents a hydrophobicity scale of nonpolar amino acids calculated by the method of Tanford. The evaluation of stability of hydrophobic cores of globular proteins using this hydrophobicity scale is described. Stereochemical and thermodynamic parameters are presented that make it possible to choose replacements for side chains of nonpolar amino acid residues in the hydrophobic cores in order to increase their stability. PMID- 9864469 TI - L-Arabinose induces synthesis of secreted beta-galactosidase in the filamentous fungus penicillium canescens AB - The mechanism of induction of secreted beta-galactosidase was studied in the filamentous fungus Penicillium canescens. L-Arabinose and its metabolite L arabitol induce the synthesis of the enzyme. Apart from beta-galactosidase, L arabinose induces the synthesis of other extracellular carbohydrolases including alpha-L-arabinofuranosidase. Increasing L-arabinose concentration above 1 mM or addition of other carbon sources results in carbon catabolite repression of the synthesis of the secreted enzymes. The data suggest that arabinofuranosidase can regulate the synthesis of secreted enzymes in P. canescens, thus controlling the level of free L-arabinose. PMID- 9864470 TI - Regulation of activity of chloroperoxidase from Serratia marcescens. AB - The influence of various factors on the activity of chloroperoxidase from Serratia marcescens was investigated. The enzyme is active only in acetate containing buffers within the pH range 4.2-5.8. F-, Cu2+, [Fe(CN)6]4+, and [Fe(CN)6]3+ inhibit the enzyme. The chloroperoxidase is thermostable and resistant to the effect of lower alcohols. PMID- 9864471 TI - Dissociation of creatine kinase under different denaturation conditions. AB - Dissociation of dimeric creatine kinase under different denaturation conditions was investigated using the cleavable cross-linker 3, 3;-dithiobis(succinimidyl propionate). The results show that at low denaturant concentrations or at low denaturation temperatures the creatine kinase was mostly inactivated, but the enzyme was still either in the dimeric state or very slightly dissociated. It appears, therefore, that for several denaturation conditions, inactivation of the enzyme is not due to the dissociation of the active dimer. PMID- 9864472 TI - Inactivation and conformational changes of yeast alcohol dehydrogenase in trifluoroethanol solutions. AB - Conformational changes of yeast alcohol dehydrogenase in trifluoroethanol solutions have been followed by fluorescence emission and circular dichroism spectroscopy. At low concentration (less than 5%), trifluoroethanol shows a reversible inhibition competitive to ethanol and noncompetitive to NAD+. The inhibition constants for native and structural-zinc-removed yeast alcohol dehydrogenase were 5.8 and 1.1 mM, respectively, suggesting that the active site becomes more flexible after the structural zinc is removed. At higher trifluoroethanol concentrations the enzyme was irreversibly inactivated. Comparison of inactivation and conformational changes of yeast alcohol dehydrogenase denatured in trifluoroethanol solutions shows that the extent of inactivation is larger than the extent of conformational changes at the same trifluoroethanol concentration. The results obtained from circular dichroism spectra show that the presence of trifluoroethanol can induce the formation of secondary structure of the enzyme. PMID- 9864474 TI - Biosynthesis of chlorophyll P-680 of photosystem II in greening leaves of plants adapted to heat shock AB - In etiolated pea and maize leaves illuminated after incubation at 38 degreesC, a new dark reaction was shown manifested in the bathochromic shift of spectral bands and accompanied by esterification of the product of protochlorophyllide photochemical reduction--Chld 684/676: Chld 684/676 --> Chl 688/680. After completion of the reaction a rapid (20-30 sec) quenching of the fluorescence of the reaction product (Chl 688/680) was observed. The reaction Chld 684/676 --> Chl 688/680 is inhibited under anaerobic conditions and in the presence of cyanide; the reaction accompanied by Chl 688/680 fluorescence quenching is not observed in pea mutants with impaired function of photosystem II reaction centers. The spectral properties of the formed Chl form with the absorption maximum at 680 nm, fluorescence quenching, and simultaneous synthesis of pheophytin suggest that the reaction is connected with the chlorophyll of photosystem II reaction center--P-680. PMID- 9864473 TI - Noncovalent complexes between poly(ethylene glycol) and proteins. AB - A new method of formation of noncovalent complexes between poly(ethylene glycol) (PEG) and proteins (alpha-chymotrypsin (ChT), lysozyme, bovine serum albumin) under high pressure has been developed. The existence of polymer in complexes was proved using 3H-labeled PEG. Complexes between PEG and ChT were studied in detail. It was shown that the composition of complexes (the number of polymer chains per ChT molecule) depends on the molecular mass of PEG and decreases with the increase of molecular mass from 300 to 4000. At the same time, the portion of the protein (wt. %) in complexes does not depend on the molecular mass of incorporated PEG and corresponds to approximately 70 wt. %. It was shown that kinetic constants for enzymatic hydrolysis of N-benzoyl-L-tyrosine ethyl ester and azocasein catalyzed by the PEG-ChT complexes are identical to the corresponding values for the native ChT. The conformational properties of ChT in complexes were studied by circular dichroism. It was shown that the enzyme in complexes fully retains its secondary structure. The estimation of steric availability of PEG polymer chains in complexes was evaluated by the complexation with alpha-cyclodextrin (CyD). It was shown that in contrast to free PEG, only part (approximately 10%) of PEG polymer chains in PEG--ChT complexes participate in the complexation with CyD. Hence, the complexation of PEG with ChT proceeds by means of multipoint interaction with surface groups of the protein globule in a region far from the active site of the enzyme and results in the significant decrease in the mobility of polymer chains. PMID- 9864475 TI - An equation for the reconstitution of membrane proteins and a method for determination of pore content in biological membranes. AB - The dependence between specific trapped volume of liposomes (W) and protein concentration (p) is proposed to be used for quantitative pore determination in biological membranes via pore reconstitution into liposomes. This dependence is described by the following equation: p = -p(e) x ln(W/W0), where W0 is initial trapped volume of liposomes and pe is an equivalent protein concentration at which molar concentrations of pores and liposomes become equal. Experimentally determined equivalent protein concentration pe is the basis of the method. This method also permits determination of molar mass of pore-forming complexes provided that preparations contain purified complex. PMID- 9864476 TI - Biochemical mechanisms of evolution and the role of oxygen. AB - The concept formulated here presumes the existence of specific mechanisms of evolution that save intermediate (and therefore imperfect) forms of organisms from elimination by natural selection. A change in the life strategy made in situations when the appearance of a new trait worsens, rather than improves, adaptation of the organism to the changing environment can be one of these mechanisms. The concept postulates that, in such cases, K-strategy (relatively low rates of reproduction and activity in general but long life span) is replaced by r-strategy (high activity and reproduction but short life span). A decrease in the life span upon the K --> r transition is suggested to be an unavoidable consequence of an elevation of formation of toxic reactive oxygen species under conditions of increased rates of aerobic metabolism required for the increased life activity. The phenomenon of giantism of transgenic tobacco plants that overproduce a mitochondrial heat shock protein (experiments done by A. Moore) is assumed to be explained by an r --> K transition. On the other hand, a decrease in the life activity and a considerable increase in life span occurring in a nematode upon mutations inhibiting the CoQ biosynthesis (S. Hekimi) might serve as an example of a K --> r transition. PMID- 9864477 TI - [Fever in intensive care: keep medications in mind at all times]. AB - In two patients, men aged 35 and 69 years admitted postoperatively to the intensive care unit, fever of unknown origin developed. One had been admitted because aspiration was suspected. He had been treated immediately with amoxicillin and clavulanic acid. The other had undergone oesophageal excision and gastric reconstruction because of oesophageal carcinoma and had been subjected to antibiotic decontamination (amphotericin B, norfloxacine en fungizone). No cause for the fever was detected, but it quickly subsided after discontinuation of the amoxicillin-clavulanic acid and the norfloxacine, respectively. When encountering fever of unknown origin in intensive care patients it is always important to think of drug fever. PMID- 9864478 TI - [Unconventional imaging techniques in inflammatory bowel diseases]. AB - In patients with inflammatory bowel disease (IBD), radiologic examinations are important for diagnosis and treatment. With conventional X-ray examinations, mucosal abnormalities, ulcers and fistulas can be visualised, but no information on the extramural extension of the disease can be obtained. Newer radiologic modalities (ultrasound, CT and MRI) offer new diagnostic possibilities. With ultrasound IBD can be diagnosed with good confidence and it can differentiate between Crohn's disease and ulcerative colitis. CT and MRI are indicated not so much to diagnose the disease but rather to determine the severity and spread of disease activity (transmural and extramural inflammation) and to detect complications such as fistulas and abscesses. PMID- 9864479 TI - [The drawbacks of diagnostic diligence]. AB - More, earlier and better diagnostic work is being done nowadays, leasing to detection of abnormalities and preliminary stages that used to remain undetected; a large reservoir of subclinical disorders is found to exist. More intensive and sensitive diagnostic methods as a rule lead to higher disease prevalence figures, with the consequences of a seeming increase of disease risk, unnecessary further examinations, treatment and follow-up of individuals and overestimation of the effects of treatment. This may even start a vicious circle. More attention should be given to using diagnostic methods in such a way that the earlier and more frequent detection of disease actually profits the patient. The proof of this should be found in scientific (population) studies of the magnitude and severity of the burden of disease, the determinants of progression, the severity of abnormalities and diseases and the favourable effect of (early) treatment. PMID- 9864480 TI - [Late sequelae of oncologic treatment in children]. AB - The risk of late effects of cancer treatment in children is higher than that after treatment during adulthood. The late effects of chemotherapy are proportional to the dosage and those of irradiation to the size of the radiation field, fractionation and dose. Irradiation may lead to impaired growth of bone and soft tissues, cranial irradiation to pituitary deficiencies, alopecia and impaired cognitive function, irradiation of the neck to altered thyroid function, thoracic irradiation to diminished pulmonary function and cardiovascular morbidity, radiation therapy of the abdomen to infertility in females, impaired renal function and chronic enteritis. Chemotherapy-induced damage is more organ specific. Well-known cardiotoxic agents are the anthracycline derivatives. Restricted pulmonary function is seen after treatment with bleomycin and nitrourea derivatives. Several antineoplastic agents are gonadotoxic in men. Nephrotoxic agents are cisplatin and ifosfamide. The cumulative relative risk of developing a second primary neoplasm is 3,8-6,9 after a follow-up period of 25 years. Alkylating agents and the topoisomerase inhibitors are known to increase the risk of haematologic malignancy, while radiation therapy is associated with bone, soft tissue, thyroid, breast, brain and gastrointestinal malignancies. PMID- 9864482 TI - [Prone position favors motor development of infants]. AB - OBJECTIVE: To determine the motor development in infants sleeping in the supine position compared to infants sleeping in the prone position, and to compare the levels of motor development of infants playing only in the supine position and of infants playing in the prone position as well. DESIGN: Prospective blinded comparing study. SETTING: Department of Physical Therapy, Maasziekenhuis, Boxmeer, the Netherlands. METHODS: Various measuring instruments were used in the home environment to determine the levels of motor development at the age of 5 months of 21 healthy infants born at term selected from a group of 160 infants attending the infant welfare clinic. RESULTS: Infants sleeping in the prone position (n = 8) showed a higher motor development than infants sleeping in the supine position (n = 13). Infants playing in the prone and supine position (n = 5) had a higher motor development than infants who played exclusively in the supine position (n = 15). CONCLUSION: Sleeping and playing in the prone position was accompanied by a higher motor development in healthy mature-born infants at the age of 5 months. PMID- 9864481 TI - [Treatment results and the prognosis in patients with localization of non Hodgkins-lymphoma in the central nervous system]. AB - OBJECTIVE: A description of clinical features and treatment results in patients with a central nervous system (CNS) localization of systemic non-Hodgkin's lymphoma (NHL). DESIGN: Retrospective. SETTING: Department of Neuro-oncology and Haematology, Daniel den Hoed Cancer Centre, Rotterdam, the Netherlands. METHODS: All patients with NHL (but not primary CNS lymphoma) treated in the period January 1st, 1990-December 31st, 1996 at the department of neuro-oncology were reviewed for presence of CNS localizations. Clinical characteristics like malignancy grade, disease stage, presence of extranodal localizations, B symptoms, serum LDH, neurological signs and symptoms, results of ancillary investigations, treatment and response were recorded. RESULTS: In this period 25 patients were diagnosed with leptomeningeal lymphoma, 2 with an intracerebral lymphoma, and 7 with both. In almost all patients the CNS localization developed during systemic progression of the NHL. Most patients presented with a radicular syndrome, cranial nerve deficits, headache or encephalopathy. More than 80% of the patients showed clinical improvement after treatment with intrathecal chemotherapy, radiation therapy or a combination of both. The median survival was three and a half months, six month survival was 32%. Progression of systemic disease was the most frequent cause of death. CONCLUSION: In most patients good palliation of neurological signs and symptoms could be obtained with intrathecal chemotherapy and radiation therapy. Survival in these patients was limited, however, in part because of the frequent concurrent progression of systemic disease. Patients with CNS localizations with chemotherapeutic treatment possibilities with a realistic chance of cure or longterm survival should be treated accordingly. Otherwise, treatment should consist of intrathecal chemotherapy or radiation therapy of involved areas only. PMID- 9864483 TI - [Breast cancer after radiation treatment for Hodgkin disease]. AB - Mammary carcinoma was diagnosed in two women aged 31 and 38 years after they had been subjected to mantle irradiation for Hodgkin's disease at the ages of 17 and 20 years, respectively. Breast amputation was performed in one case, lumpectomy in the other; the subsequent courses were uncomplicated. During follow-up there was no sign of recurrence. Lifelong follow-up by specialists of women irradiated for Hodgkin's disease should be considered; monthly self-examination of the breasts should be advised and mammography should be carried out annually as from 10 years after thoracic or axillary irradiation. It is especially women treated before their 20th year of life who run a relatively high risk. PMID- 9864484 TI - [Inhalation therapy in children younger than 2 years]. PMID- 9864485 TI - [Inhalation therapy in children younger than 2 years]. PMID- 9864486 TI - [Inhalation therapy in children younger than 2 years]. PMID- 9864487 TI - [Misunderstanding of disciplinary decisions]. PMID- 9864488 TI - [Misunderstanding of disciplinary decisions]. PMID- 9864489 TI - [Misunderstanding of disciplinary decisions]. PMID- 9864490 TI - [The family physician and the postcoital test]. PMID- 9864491 TI - [Acute urine retention in women]. PMID- 9864492 TI - [Acute urine retention in women]. PMID- 9864493 TI - [The treatment of subglottic laryngitis (pseudocroup): steroids instead of steam]. PMID- 9864494 TI - [The bikini incision: nice, but not without painful complications]. PMID- 9864495 TI - [The relationship between serum prostate-specific antigen and the skeletal- and CT-scan in staging of primary prostate cancer]. PMID- 9864496 TI - [NSAID-use delays the diagnosis of 'acute abdomen']. AB - In three patients with abdominal pain, two men aged 63 and 18 years and a woman aged 46 years, the use of NSAIDs reduced the symptoms. They were in fact suffering from peritonitis due to gastrointestinal perforation, but the decision to operate was delayed because of the relatively mild presentation. The strong analgesic, antipyretic and anti-inflammatory properties of NSAIDs can reduce the symptoms, signs and laboratory findings of peritonitis. PMID- 9864497 TI - [The sentinel node procedure: standard intervention for surgical treatment of breast cancer]. AB - Axillary lymph node dissection may be avoided in the surgical treatment of breast cancer if the sentinel node appears to be free of metastatic tumour cells. The sentinel node concept has been validated in hundreds of patients, in whom this node has been localized successfully by dye-guided node mapping and (or) gamma probe localization after injection of a radiopharmaceutical agent. The success rate of sentinel node localization depends on different factors, such as type, volume and injection site of the radiopharmacon, choice of the handheld gamma probe, timing of the blue dye injection, and clinical stage of the disease. The combination of preoperative lymphoscintigraphy, intraoperative gamma-probe guidance and blue dye administration will increase the success rate of a sentinel node biopsy. Recent reports, from both Europe and the United States, appear to indicate that the sentinel node biopsy will soon be standard procedure in the surgical treatment of (T1-2) breast cancer. In the majority of breast cancer patients axillary lymph node dissection, a cause of much morbidity, may thus be avoided. PMID- 9864498 TI - [The sentinel node biopsy in patients with breast cancer: recommendations for the introduction of this procedure. Dutch Working Group Sentinel Node Biopsy in Breast Cancer]. AB - Sentinel node biopsy may be useful in the staging of breast cancer. In experienced hands presence or absence of metastasis in the sentinel node accurately predicts the nodal status and in patients with a negative sentinel node biopsy axillary dissection may be avoided. The technique, however, shows a clear learning curve and hasty introduction may lead to an unacceptable rate of false negative biopsies. Moreover, the introduction of the technique in general practice should be well coordinated in order to prevent large differences between hospitals in the staging and treatment of breast cancer patients. On the initiative of NABON (Nationaal Borstkanker Overleg Nederland; 'national breast cancer consultation') and the Nederlandse Vereniging voor Chirurgische Oncologie (Netherlands Association for Surgical Oncology) a study group was formed to co ordinate the introduction of the sentinel node biopsy in breast cancer patients in the Netherlands. Important topics are central registration of all procedures in order to evaluate its applicability and technical variations, the performance in each centre of at least 50 procedures together with axillary node dissection to evaluate the results before clinical application is considered, and nationwide co-ordination to find the best therapy for patients with positive sentinel nodes. PMID- 9864499 TI - [Positive developments for patients with breast cancer as a result of the sentinel node biopsy procedure]. AB - As an increasing number of large prospective studies show a high accuracy of the sentinel node for the staging of the axilla in women with invasive breast cancer, there is no need to test the value of this new technique in a randomised trial. Much more emphasis should be given to a reliable implementation of the technique in general practice, requiring a closely co-operating multidisciplinary team meticulously performing the different steps of the technique. The guidelines designed by the Dutch Working Group on the Sentinel Node in Breast Cancer include a learning phase of--arbitrarily--50 procedures in patients also undergoing a complete axillary dissection. What remains is the need for a treatment trial aimed at reducing the morbidity of the treatment of axillary metastases while retaining equal regional tumour control and patient survival. As the indication for adjuvant systemic treatment has shifted from the N-stage to T-stage parameters (size, grade, mitotic activity), the axillary nodal status has become less important as a staging tool. Thus, comparison of surgery or radiotherapy of sentinel node positive patients in a randomised trial appears to be a logical next step. PMID- 9864500 TI - [Infections and aging]. AB - Elderly people are exposed to infectious risks in ways different from younger people. These risks relate to environment, transmission routes and vectors, microbial colonisation of body surfaces and quality of host defence. Admission to a nursing home or a hospital may easily lead to colonisation with (resistant) potentially pathogenic micro-organisms, while pre-existing morbidity, medication and functional disability impair the quality of host defence, to a greater degree than the process of aging. In general, infections have a great impact on the elderly patient. Not only are infections in the elderly often serious, but they often have a deleterious effect on comorbidity and functional ability. Infections in elderly people with serious comorbidity face the physician with the dilemma how far to go with diagnostic procedures and therapy. PMID- 9864501 TI - [Roaming through methodology. IX. The interpretation of subgroup analyses]. AB - The results of subgroup analyses can be very informative, in particular for clinicians interested in evidence pertaining most directly to a particular patient. However, subgroup analyses may also be potentially misleading. The strength of evidence for subgroup effects depends on the question whether hypotheses have been defined prior to analysis, whether potential problems regarding multiple comparisons have been considered and whether there is biological plausibility of the effects. Using these guidelines the reader of a trial report should be able to decide if presented subgroup effects are of clinical importance or if the overall result is a better estimate of treatment effect. PMID- 9864502 TI - [Guideline 'Radiotherapy in non-small-cell lung carcinoma. Working Group, National Organization for Quality Assurance in Hospitals, Netherlands]. AB - 8000-9000 new patients with lung cancer are diagnosed yearly in the Netherlands. Of these 10-30% can be treated by surgery, which because of co-morbidity or the extent of the disease is impossible in the others. There is agreement on the terms 'curative', 'radical' and 'palliative' radiotherapy: curative radiotherapy aims at cure through destruction of all tumour cells, on the assumption that the neoplasia has not metastasized (this applies to patients with tumours in stage I and II and sometimes in stage III); radical radiotherapy aims at postponement of locoregional tumour growth; this has a positive effect on the patient's duration of survival and quality of life, but it is to be expected that the patient will die from distant metastases (this treatment is indicated for patients with stage IIIA disease); palliative radiotherapy aims at improvement of the quality of life rather than on prolonging the duration of survival. Patients with a stage III tumour (locoregional invasion making surgical treatment impossible but without distant metastases) without complaints should receive radiotherapy, because these patients are candidates for prolonged survival. In the future the best therapy for these patients could be a combined modality (chemotherapy in combination with radiotherapy). Lung cancer patients with an irresectable tumour should be included as often as possible in clinical trials. During treatment checking quality of life, taking into account the objective and the side effects of the treatment, is important. All medical experts who look after the patient, should inform each other on the actual state of health of the patient and any appointments made. A personal file with this information, kept by the patient himself, has been advocated for this purpose. PMID- 9864503 TI - [A Dutch family with an autosomal dominant form of diabetes mellitus as a result of a mutation in the HNF1 alpha-gene (MODY3)]. AB - OBJECTIVE: To study the molecular genetic basis of an autosomal dominant form of diabetes mellitus in a Dutch family. DESIGN: Descriptive. SETTING: Academic Hospital Nijmegen, the Netherlands and Laboratories for Molecular Genetics, Birmingham and Exeter, Great Britain. METHODS: A large pedigree with maturity onset diabetes of the young (MODY) was studied by taking a history and by performing laboratory analysis; DNA was isolated from peripheral blood lymphocytes and linkage analysis was carried out using genetic markers near known MODY loci. As linkage to the MODY3 gene (encoding for the transcription factor hepatocyte nuclear factor (HNF)1 alpha) was suggested, all exons of the gene were sequenced for mutation detection. RESULTS: Of the 27 family members 13 were affected by diabetes. Diabetes was diagnosed at a mean age of 36.7 years, with four family members diagnosed before the age of 25. Nearly all patients were treated with diet/oral antidiabetics. Diabetic family members had lower fasting (specific) insulin concentrations than normoglycaemic family members (53.8 (SD: 5.4) versus 90.4 (SD: 12.9) pmol/l; p < 0.05). Linkage to the MODY3 gene was established. Further investigation showed a mutation in exon 2 of the gene. CONCLUSION: This Dutch family suffered from an autosomal form of diabetes mellitus caused by a mutation in the HNFI alpha gene (MODY3). Phenotypically, the diabetes appeared to be relatively mild; it was characterised by impaired insulin secretion. PMID- 9864504 TI - [A twelve-year old girl with diabetes mellitus type 2]. AB - A 12-year-old very obese girl was referred for hyperglycaemia. She had no complaints apart from a recent vaginal candidiasis. Non-insulin-dependent diabetes mellitus (type 2 diabetes) was diagnosed. She was started on a hypocaloric diet and on an oral hypoglycaemic agent (metformin 500 mg/day). This case illustrates the importance of awareness of the existence of type 2 diabetes in childhood and adolescence. PMID- 9864505 TI - [The hyperglycemic dehydration syndrome]. PMID- 9864506 TI - [The hyperglycemic dehydration syndrome]. PMID- 9864507 TI - [The hyperglycemic dehydration syndrome]. PMID- 9864508 TI - [Secondary amenorrhea due to intrauterine adhesions: Asherman's syndrome]. AB - Three women aged 35, 33 and 40 years had secondary amenorrhoea and monthly recurring abdominal pains after curettage for blood loss after a pregnancy. Hysteroscopy revealed intrauterine adhesions. The symptoms disappeared largely or completely after synechiolysis, introduction of an IUD and oestrogen treatment. Changes in the menstrual cycle, infertility or recurrent abortions starting after a puerperal or postabortum curettage should cause the physician to suspect intrauterine adhesions. For the diagnosis hysteroscopy is the method of choice and hysterography should be added for preoperative assessment and classification. Blind procedures for rupturing the adhesions should not be performed because of the risk of perforation and creating false routes. PMID- 9864509 TI - [The erection pill]. AB - The introduction of sildenafil (Viagra) for the treatment of erection disorders has received much international attention. In the USA the prevalence of severe erection disorder amounts to 10% for men aged 40-70 years, which is much higher than was expected, in the Netherlands as well. This is probably due to embarassment with the condition and the absence of simple treatment. With the advent of sildenafil the problem of erection disorder is now openly discussed. An interview investigation in Leiden, the Netherlands, showed that two-thirds of men with erection problems (n = 200) who volunteered to participate in a relevant study, had consulted to a physician before; one-third of the volunteers were interested in using an erection pill, one-third also in other treatments, and one third wanted to be treated by any method. These figures indicate that most men already take their problem seriously. As age is the main contributing factor to erection disorder, the safety of sildenafil in the elderly is a point of concern. Nitrate use is an absolute contraindication, poor cardiovascular condition is a relative contraindication. Sildenafil will probably become the drug of choice for the treatment of erection disorder. It constitutes a major step forward. PMID- 9864510 TI - [Somatization of erectile disorders by the introduction of sildenafil]. AB - With the introduction of the erection-supporting oral drug sildenafil (Viagra) the development of somatic treatments of male sexual dysfunction has reached a temporary peak. The availability of a medicament that is easy to take will result in an increase of the incidence and possibly also the prevalence of erectile disorder. It may even lead to the myth that only a perfect erection is normal. The question is where to draw the line between a normal inconvenience of life and a serious disease. The erection pill will probably be preferred to all other treatments. Therefore, most patients will consult their family physician, who will refer only difficult cases to urologists or sexologists. Physicians should take 'the man behind the penis' into consideration before writing a prescription, otherwise a distressed man without an erection might become a distressed man with an erection. In the long run, psychosexual therapy is less expensive than taking sildenafil for many years. PMID- 9864511 TI - [Sildenafil (Viagra) for the treatment of erectile disorder]. AB - Erectile dysfunction is a common but underreported condition. It is to be expected that the number of patients consulting their physician with the complaint of erectile dysfunction will increase considerably with the introduction of sildenafil (Viagra), the first oral drug that enhances penile erection. Sildenafil is an inhibitor of the enzyme phosphodiesterase type 5. It causes erection of the penis by allowing the relaxation of the smooth musculature of the cavernous body to persist. The first clinical results indicate that the treatment with sildenafil is safe and effective in the hands of a sexologically qualified physician. An erection disorder is essentially not more than a symptom which primarily requires causal therapy. PMID- 9864512 TI - [Endoscopic treatment of Barrett esophagus]. AB - The purpose of the treatment of Barrett epithelium in the distal oesophagus is to reduce or even eliminate the increased risk of malignant degeneration in it. This can be achieved by removing the Barrett epithelium, whether or not dysplastic, and to have it replaced by normal squamous epithelium. Drug treatment or surgical antireflux treatment of Barrett epithelium has hardly any effect on the length of the Barrett epithelium or on the occurrence of malignancy. Various forms of endoscopic ablative therapy (laser coagulation, multipolar electrocoagulation, photodynamic therapy and argon plasma coagulation), in combination with antireflux treatment enable removal of the Barrett epithelium with regeneration of squamous epithelium. However, islets of Barrett epithelium may be found beneath the regenerated squamous epithelium and there is also the possibility of malignant potential of pluripotent stem cells left behind in the oesophagus. Future studies will have to afford insight into long-term results, the costs, the side effects of the various methods of treatment and the quality of life of patients during and after treatment of the Barrett oesophagus. PMID- 9864513 TI - [Roaming through methodology. X. The testing of diagnostic procedures]. AB - In this era of evidence-based medicine, diagnostic tests cannot escape close scrutiny of their effectiveness. Sensitivity and specificity have up till now played a central role in the evaluation of diagnostic tests. These terms are not without their shortcomings when it comes to the characterisation of a test's true worth for patients. Randomised clinical trials are increasingly used for evaluation of medical tests and outlining of strategy. The indirect relationship between test results and health outcome creates additional challenges for designers of such trials. PMID- 9864514 TI - [A patient with tetanus without an obvious point of entry]. AB - A 59-years-old man with oesophageal cancer (T3NXMo) presented with trismus, dysarthria and diaphoresis. Later, he developed opisthotonus and generalized spasms. Despite negative blood cultures and sufficiently high anti-tetanus titres, tetanus was suspected, on clinical grounds. He was intubated and treated with tetanus toxoid, human antitetanus immunoglobulin, benzylpenicillin, propofol, benzodiazepines, vecuronium, and sufentanil, and recovered gradually. Tetanus is caused by Clostridium tetani, a Gram-positive rod capable of remaining present latently in the body for years. Absence of a visible external wound suggests that the oesophageal mucosal cancer lesion could have served as portal of entry or that endogenous reactivation of latent tetanus bacteria had taken place. PMID- 9864515 TI - [The epidemiology of hepatitis A in the Netherlands, 1957-1998]. AB - OBJECTIVE: To describe the incidence of hepatitis A in the Netherlands, 1957-1997 to find an explanation for the increase in 1998. DESIGN: Descriptive. SETTING: National Institute of Public Health and Environment (RIVM), Bilthoven, the Netherlands. METHOD: Following an increase of the number of hepatitis A notifications in 1998 the national notification data over the period 1957-1997 were examined. The impact of waning natural immunity in the Dutch population and of the annual influx from high-endemic countries on the epidemiology of hepatitis A was studied. RESULTS: The number of notifications declined considerably in the early sixties and stabilised in the seventies at 5-7 notifications per 100,000 inhabitants. The age distribution over time showed that the proportion of cases in children is still the largest, although the proportion of notifications of adult cases increased slightly over time. A seasonal fluctuation was demonstrated by a steep increase in the number of notifications among non-Dutch children in autumn, with the infection acquired abroad, probably in Morocco or Turkey, followed by an increase among children and adults who acquired their infection in the Netherlands. The number of notifications doubled in the early months of 1998 compared with 1997. The present epidemic increase can be explained by increased transmission in subgroups who contributed similarly in the same months in the past 5 years and may result from secondary infections related to the relatively high peak in autumn in 1997. CONCLUSION: In 1975-1997 a stable but low incidence of hepatitis A was observed in the Netherlands; the number of cases among adults did not increase over time. The present epidemic increase underlines the potential risk of epidemics and the role of importation of the virus from high endemic countries. PMID- 9864516 TI - [The risk of Kahler disease (multiple myeloma) in patients with a paraproteinemia: myeloma risk score, developed in the region of the Integral Cancer Center, West]. PMID- 9864517 TI - [The risk of Kahler disease (multiple myeloma) in patients with a paraproteinemia: myeloma risk score, developed in the region of the Integral Cancer Center, West]. PMID- 9864518 TI - [Obesity]. PMID- 9864519 TI - [Revised consensus 'Diagnosis of dementia syndromes']. PMID- 9864520 TI - [Malignant or nonmalignant pericarditis]. AB - Pericardial effusion can occur as the first manifestation of disseminated cancer, as was demonstrated in the case of a 59-year-old woman who presented with respiratory insufficiency. In contrast, in three breast cancer patients aged 44, 55 and 53 pericardial effusion was shown to have a non-malignant cause, such as radiation or chemotherapy. Careful diagnostic procedures, including pericardiocentesis, should be performed to adjust the treatment to the underlying cause and to optimize the prognosis. It should be kept in mind that pericarditis in about half the patients suffering from a malignancy has a non-malignant cause. PMID- 9864521 TI - [Diagnostic imaging techniques in psychiatry]. AB - Morphological and functional abnormalities have been found in several psychiatric diseases, including schizophrenia, schizophreniform disorder, manic-depressive disorder, obsessive compulsive disorder, posttraumatic stress disorder, and panic disorder. Most research concerned schizophrenia. Altered volumes of the ventricles, cortical gray matter, basal nuclei, thalamus and limbic structures have been reported. Altered activity was found in several cortical and subcortical brain structures, in particular the frontal cortex, during cognitive and pharmacological challenge. Disease progress, genetic and environmental factors appear to influence the brain abnormalities. The specificity of the findings is still limited. The use of imaging techniques in the psychiatric clinic is still restricted. Notwithstanding these limitations, focus in psychiatry research has been switched from the question if brain abnormalities exist to where they are, of what they consist and where they originated. PMID- 9864522 TI - [Multiples births: a continuing problem with assisted reproductive techniques]. AB - Assisted reproductive techniques (ART) such as in vitro fertilisation (IVF), ovulation induction and superovulation followed by insemination have caused a sharp increase in multiple birth prevalence rates. The perinatal morbidity of multiple birth infants is high because of the high incidence of premature birth. The social and psychological problems of multiple birth families are also considerable. In 1990 high-order multiple births were mainly the result of IVF. Although the number of IVF treatments has increased more than the number of other ART treatments, nowadays high-order multiple births are predominantly caused by superovulation. Since 1990 the number of high-order multiple births has stabilised, but the recent sharp increase in ART twins results in a further rise in births of severely preterm ART infants. More restraint should be practised in superovulation treatments to bring down the number of (high-order) multiple births. PMID- 9864523 TI - [Drowning and near-drowning in children]. AB - Drowning and near-drowning are major causes of death and neurological damage, respectively, in children. The pathophysiological substrate consists of hypoxia, ischaemia, respiratory and metabolic acidosis and sometimes, hypothermia. Most cases involve aspiration of liquid; this leads to a persistent impairment of the gas exchange. Occurrence of arrhythmias and hypovolaemia is very likely. The main objective of treatment of the near-drowned is limiting cerebral damage. Treatment consists of resuscitation and stabilization, administration of oxygen with positive end-expiratory pressure, intravenous administration of liquids and central reheating. The prognosis depends in the first place on the duration of the submersion, which, however, is often difficult to establish. Submersion for over 5 minutes is prognostically unfavourable. In hypothermia due to submersion in ice cold water the prognostic factors are less clear--in these cases the treatment should always be continued until the core temperature is > or = 32 degrees C. PMID- 9864524 TI - [The management of acute hepatitis C and needlestick accidents with hepatitis-C virus positive blood]. AB - Acute hepatitis C is rarely diagnosed, in part because of its usually subclinical course. Infection with the hepatitis C virus (HCV) has a high chronicity rate, 70 90%. The risk of infection after a needlestick accident with HCV positive blood is 3-10%. There are no efficacious preventive measures regarding HCV infection but treatment with the antiviral drug interferon alpha during the acute phase of the disease has shown to significantly reduce the risk of subsequent chronic infection. It is advised to evaluate individuals who were exposed to infected blood by a needlestick accident regarding HCV transmission, and to offer interferon treatment to them in case they become HCV positive, as demonstrated with a positive serum HCV-RNA test. PMID- 9864525 TI - [Elective cesarean section is preferred after the completion of a minimum of 38 weeks of pregnancy]. AB - OBJECTIVE: To determine the frequency of neonatal respiratory morbidity following elective caesarean delivery at term and to identify prognostic factors for this morbidity. DESIGN: Retrospective. SETTING: Academic Hospital Utrecht/Wilhelmina Children's Hospital Neonatal Intensive Care Unit, Utrecht, the Netherlands. METHOD: All elective caesarean deliveries in the Academic Hospital Utrecht from the period 1990-1995 were studied. Also, neonates were included who were admitted for intensive neonatal care because of respiratory insufficiency following elective caesarean delivery in the region surrounding Utrecht. An elective caesarean delivery was defined as a delivery performed after 37 weeks of gestation without any complicating factor that might influence the timing of delivery. Prognostic factors for neonatal morbidity after caesarean delivery were identified by multivariate logistic regression analysis. RESULTS: During 1990 1995, 272 elective caesarean deliveries after 37 weeks of gestation were performed that fulfilled the inclusion criteria; 5.1% of the neonates were admitted to the medium care unit because of respiratory problems. The relative risk of respiratory morbidity after delivery by caesarean section with a gestational age of 39-42 weeks compared with a gestational age of 37-38 weeks, was 0.14 (95% confidence interval: 0.03-0.64; p < 0.001). Male sex was a cofactor. Nine neonates of whom 8 (90%) had a gestational age of less than 39 weeks were admitted to the intensive care unit. CONCLUSION: Most of neonatal respiratory morbidity could have been avoided by postponement of the at-term elective caesarean section until a certain gestational age of at least 38 complete weeks. An elective caesarean section should not be performed before that period. PMID- 9864526 TI - [Discrepancy between the guidelines and practice by family physicians in treating adults with an exacerbation of asthma or chronic obstructive pulmonary disease]. AB - OBJECTIVE: To analyse to what extent current management of exacerbation in adult asthmatics and patients with asthma or chronic obstructive pulmonary disease (COPD) in general practice is consistent with the guidelines from the standard 'asthma/COPD in adults' of the Dutch College of General Physicians (NHG). DESIGN: Cross-sectional, prospective. SETTING: Centre for Quality of Care Research, University of Nijmegen, the Netherlands. PATIENTS AND METHODS: Fifty-two GPs in 1992 during an average of 3.5 months in adult patients with exacerbation of asthma or COPD prospectively registered the severity of the dyspnoea, productiveness of coughing, colour of phlegm, fever, auscultatory findings, patients' age and sex and use of medication. Current management (peak flow measurement, prescriptions) was related to all these data and compared with that recommended in the current NHG standard. RESULTS: Data were available on 383 exacerbations. Peak flow measurement was used diagnostically in 27% of the exacerbations. Inhalation steroids were prescribed in 26% of the cases (new or step-up), oral steroids were prescribed in 24%, more often in exacerbations with severe dyspnoea (48%; p < 0.0005), in patients who prior to the exacerbations had been using two or more different COPD drugs (37%; p < 0.0005) and in patients over 55 years of age (35%; p < 0.0005). Antibiotics were prescribed in 60% of the cases, and more often in case of coloured phlegm (83%; p < 0.0005) and of fever (91%; p < 0.0005). CONCLUSION: Objectivation of the bronchial obstruction by peak flow measurement was done less often than recommended in the standard; antibiotics were prescribed more often and steroids less often. In order to reduce the substantial discrepancy between the guidelines and the actual management, implementation should be aimed at changing GPs' routines, with special attention for barriers to change. PMID- 9864527 TI - [Paroxysmal nocturnal hemoglobinuria in pregnancy]. AB - The combination of paroxysmal nocturnal haemoglobinuria (PNH) and pregnancy occurred in three women aged 29, 31 and 35 years. One patient had a spontaneous abortion twice after which she committed suicide. In the other two the pregnancy and parturition--assisted--were without complications. However, one woman developed post-partum abdominal crises. The children were born at term and healthy. PNH is a rare form of haemolytic anemia. Pregnancy causes lethal complications in 6% of the cases of PNH, while 30% of the pregnancies end in spontaneous abortion or stillbirth. Accurate monitoring of pregnancy and the puerperium is essential. PMID- 9864528 TI - [Accidental exposure to blood and the risk of transmission of virus infections for various occupational groups in Amsterdam, 1986-1996]. AB - Since 1986 the number of parenteral exposures to potentially infectious blood reported to the Amsterdam Public Health Service increases every year. The number of needlestick accidents increased significantly from 64 in 1986 to 166 in 1996 whereas the number of other exposures decreased from 59 to 44 in these years. The increase was mainly seen in nonhospital based (para)medics. A possible explanation of this increase is greater awareness of the potential infection risk with HIV, hepatitis B or C virus leading to a tendency to report more readily. This assumption is in contradiction with results of studies in hospital-based personnel where a decrease is observed as a result of educational programmes. Other explanations are a higher frequency of use of sharp instruments and (or) an increase in the workload. Out of a total of 1886 needlestick accidents in 1986 1996 one woman became HIV positive; she was deliberately infected by her ex partner who injected her with blood of an AIDS patient, and one person contracted an hepatitis C virus infection: a policeman wounded by a needle used by a drug addict. PMID- 9864529 TI - [Work capacity evaluation within Social Security]. AB - Incapacity for work is not a medical but an economic concept. The difference between the earnings somebody could have had without health problems, and the possible earnings with health problems determine the degree of incapacity for work. Social security physicians and vocational experts together judge the degree of incapacity for work in discussion with the client. The physician judges the general possibilities of the client. The vocational expert selects possible jobs the client should be able to do. The disability allowances are relatively low. Treating physicians should take into account this and other negative aspects of incapacity for work. PMID- 9864530 TI - [Clinical and toxicological aspects of the use of ecstasy]. PMID- 9864531 TI - [Urinary tract infections in infants, an insidious clinical picture]. AB - Three boys aged 4, 5 and 7 weeks drank poorly, vomited and were lethargic. There were metabolic disorders attributable to a urinary tract infection. Ultrasonography revealed anatomical anomalies. After antibiotic treatment and, if necessary, surgical correction, the patients recovered. Follow-up was uncomplicated except persisting polyuria in one of the patients. A urinary tract infection in young children is difficult to recognise because of the aspecific presenting symptoms. It can cause a severe metabolic disturbance in which hyponatraemia and hyperkalaemia develop (pseudohypoaldosteronism), combined with metabolic acidosis and polyuria. A high alertness for urinary tract infections in young children with these aspecific symptoms is needed as well as metabolic and urologic evaluation. PMID- 9864532 TI - [Surveillance of communicable diseases in Europe; more than just a means of communication among national institutes]. AB - The re-emergence of infectious diseases as a threat to the health of the public has led to the initiation of several surveillance programmes in the European community. The impact of these Europe-wide surveillance programmes, including the surveillance of antibiotic resistance, will depend on the ability of the epidemiologists involved to gather validated data that accurately reflect the trends in the occurrence of infectious diseases and of antibiotic resistances. The results of surveillance should be communicated not only between the national institutes but should also reach the medical communities and the public at large in the participating countries. PMID- 9864533 TI - [Difficult weaning from mechanical ventilation: a graduated approach and teamwork]. AB - Weaning from mechanical ventilation in patients with chronic obstructive pulmonary disease (COPD) or left ventricular failure may be difficult. At the time of intubation and initiation of mechanical ventilatory support, this treatment is usually life-saving in the short term. Only later on, the condition which necessitated such support may prove irreversible. COPD patients often require positive end-expiratory pressure to enable them to trigger the ventilator comfortably. Patients with left ventricular failure need pharmacological support to reduce the circulating volume and to reduce left ventricular afterload because the ventilatory support itself reduces both left ventricular preload and afterload. Gradual withdrawal of pressure support and gradually increasing periods of T-piece weaning are probably equally effective. New methods have been described but have not yet been tested in randomized controlled trials. More important than the method of weaning is the presence of qualified and dedicated nursing support. Not all patients can be weaned; for most of those who cannot, prospects are grim. These patients require optimal palliative support with recognition of their autonomy. PMID- 9864534 TI - [A changed pattern of opportunistic infections and malignancies in HIV seropositive patients after the introduction of intensive anti-HIV-combination therapy]. AB - The application of potent combinations of antiretroviral drugs ('highly active antiretroviral therapy' (HAART)) makes effective therapy of HIV infection feasible. Consequently, the pattern of opportunistic infections and other secondary complications has changed. The incidence of infections and mortality due to aids has declined significantly. Further, the occurrence of other infections and syndromes, till now unknown in patients with aids, has been observed. It is thought that these are caused by HAART-induced inflammation, a phenomenon due to immune enhancement following HAART. An important issue is whether primary and secondary prophylaxis against opportunistic infections can be discontinued after improvement of the immune system: indeed, there are reports that discontinuation is safe in patients with persistent CD4+ cell counts above the critical level for that particular infection while CD4+ cell counts are monitored carefully. PMID- 9864535 TI - [Changes in the energy needs of the elderly: an often encountered cause of nutritional deficiencies and frailty]. AB - Changes in the body composition of elderly people and diminished physical activity reduce the energy requirement and the daily energy intake. It is especially frail elderly people who run an increased risk of a poor nutritional condition which further enhances fragility. Frailty is often defined as the state of reduced physiologic reserve associated with increased susceptibility to disability. In frail elderly people energy requirement is often reduced. Studies have shown that when energy intake falls below a level of about 6.3 MJ, it is hard to obtain a diet with a sufficient supply of minerals and water soluble vitamins. It is especially the daily intake of vitamins of the B complex and of vitamin C that then prove insufficient. In addition, immobile elderly and elderly of 75 years and older are at risk for an inadequate vitamin D status. An adequate diet is one of the factors that may prevent frail health. Screening lists have been developed to quickly obtain information about the nutritional status of older adults. However, these lists still have to be adapted to the Dutch dietary pattern. Currently, a change of the body weight is the best warning of insufficient nutrition. PMID- 9864536 TI - [Incidental high blood pressure in family practice: due to hypertension and/or left ventricular hypertrophy in more than half of the patients]. AB - OBJECTIVE: To determine if patients with incidentally high blood pressure actually have hypertension and if these patients have an increased left ventricular mass. DESIGN: Cross-sectional study. SETTING: Two family practices with 8 general practitioners in Leiden and Noordwijk, the Netherlands. METHODS: From the Family Practice Network in the Leiden area 133 (67%) out of 200 patients with incidental high blood pressure, who did not receive antihypertensive medication, participated in the study. Their blood pressure was measured 6 times with a mercury manometer, an automatic, non-invasive ambulatory blood pressure monitoring during 24 hours was performed once and their left ventricular mass was measured by means of echocardiography. RESULTS: Of the 133 selected patients 46% had a mean diastolic blood pressure > 95 mmHg measured with the mercury manometer and 64% had a mean 24-hr diastolic blood pressure > 90 mmHg measured with the ambulatory blood pressure monitor. The correlation between both blood pressure measurements was moderate (correlation coefficient 0.73). Left ventricular hypertrophy was found in 53% of the patients, irrespective of their blood pressures. CONCLUSION: In this investigation 45-65% of patients with an incidentally high blood pressure had a mean diastolic pressure > 95 mmHg as measured with a mercury manometer and (or) a mean 24-hr diastolic blood pressure > 90 mmHg as measured with the ambulatory blood pressure monitor; 53% had left ventricular hypertrophy. PMID- 9864537 TI - [The risks from ingestion of cleaning tablets for dentures and dental prostheses]. AB - OBJECTIVE: To determine the frequency and severity of intoxication caused by cleansing tablets for dental plates or dental prostheses. DESIGN: Questionnaire study. SETTING: National Poisons Control Centre, Bilthoven, the Netherlands. METHOD: In 1996, all 78 physicians who requested information from the Centre about intoxications with cleansing tablets for dental plates or dental prostheses were sent a questionnaire enquiring about the exposure situations and the severity of the effects of the intoxications. RESULTS: Forty-six questionnaires (59%) were returned. Older people (24; 52%) and children (14; 30%) constituted the largest groups of intoxicated. As a rule, accidental ingestion of the tablets was involved. In most cases, one tablet had been swallowed. Local effects, mostly of a mild nature, were reported in 22 patients. Systemic effects were reported in two patients but in these cases the connection with the ingestion was doubtful. PMID- 9864538 TI - [A diabetic patient with cystitis emphysematosa]. AB - In a woman aged 80 years arriving in the Emergency Room with progressive malaise, anorexia and somnolence, a large resistance was found in the lower abdomen, which proved to be due to cystitis emphysematosa. The patient was known to suffer from non-insulin dependent diabetes mellitus. Imaging revealed a large accumulation of gas in the urinary bladder, which was treated successfully with catheterization and antibiotics. Cystitis emphysematosa is a rare condition, characterized by collection of gas in the bladder and bladder wall and brought about by gas forming micro-organisms that decompose glucose. In patients with diabetes mellitus optimal regulation of the blood glucose levels, with a view to preventing glycosuria, is a condition of speedy recovery. Although cases with a fatal outcome have been reported, the prognosis in general is favourable. PMID- 9864539 TI - [Hyperammonemia in hydronephrosis]. AB - A man aged 81 with bilateral hydronephrosis presented with lethargy deepening into coma caused by hyperammonaemia. The hyperammonaemia was most likely caused by excessive absorption of ammonia produced by Proteus mirabilis in the obstructed and dilated urinary tract. The patient died within a few hours. PMID- 9864540 TI - [Acute hepatocytic necrosis during ketoconazole therapy for treatment of onychomycosis. National Foundation for Registry and Evaluation of Adverse Effects]. AB - The imidazole derivative ketoconazole is approved in the Netherlands for treatment of local and systemic fungal infections. Despite the risk of major hepatic damage, this antimycotic drug is still being used in daily practice for the oral treatment of patients with onychomycosis. Since April 1986, 18 cases of relatively severe hepatic damage in the Netherlands were ascribed to the oral use of ketoconazole for the treatment of onychomycosis, skin infection or vaginal candidiasis. Given the potential risk for the patients concerned, ketoconazole should no longer be prescribed for relatively mild cosmetic disorders such as onychomycosis. PMID- 9864541 TI - [Surveillance of communicable diseases in the European Union]. AB - Important developments are taking place in the Public Health framework in the European Union (EU). A number of international surveillance networks have been developed between the countries of the EU. The European Commission has initiated horizontal and disease specific programmes in the field of infectious disease surveillance. These have stimulated European co-operation for prevention and control of infectious diseases. Much topical information can be found on several Internet pages. The challenge is to set up networks for all public health relevant infectious diseases. PMID- 9864542 TI - [The influenza season 1997/'98 and the vaccine composition for 1998/'99]. AB - The 1997/'98 influenza season in the Netherlands was marked by influenza A/H3N2 activity which never reached a true epidemic level. There was no real peak activity either but a prolonged period of increased activity of approximately eight weeks with a maximum in week 13, when sentinel physicians reported 16.6 cases of influenza-like illness per 10,000 inhabitants. It was not until week 18 of 1998 that the influenza activity declined to baseline levels. During the season, almost exclusively influenza A/H3N2 viruses were isolated, of which the majority resembled the new strain influenza A/Sydney/5/97 (H3N2). Further analysis of these variant viruses revealed that, although there was some cross reactivity with the vaccine strain (A/Nanchang/933/95), no optimal protection could be expected to be induced by the vaccine. Antigenic characterisation of the sporadic influenza A/H1N1 and influenza B viruses showed that these were related to the vaccine strains. As a result of these findings, the World Health Organization (WHO) recommended to change the H3N2 strain in the influenza vaccine for the season 1998/'99 to an influenza A/Sydney/5/97(H3N2)-like strain. Based on epidemiological data from other countries, it was also decided to change the influenza A/H1N1 component to an influenza A/Beijing/262/95 (H1N1)-like strain. PMID- 9864543 TI - [Changes in treatment based on an ECG of patients in family practice]. PMID- 9864544 TI - [Diplopia as a symptom of serious illness]. PMID- 9864545 TI - [Results of laparoscopic assisted vaginal hysterectomy in the Ikazia Hospital at Rotterdam, 1993-1997]. PMID- 9864546 TI - [Results of laparoscopic assisted vaginal hysterectomy in Ikazia Hospital at Rotterdam, 1993-1997]. PMID- 9864547 TI - [Calcitonin and stanniocalcin. Particular aspects of the endocrine regulation of phospho-calcium metabolism in mammals and fish]. AB - The hypocalcemic and hypophosphatemic peptide calcitonin (CT) is secreted by mammalain thyroid parafollicular cells and fish ultimobranchial body. Over a dozen species of CTs have been cloned and/or sequenced. They can be separated into three classes based on structural and biological similarities: teleost/avian, artiodactyl, and human/rat. In mammals, CT exerts its anti hypercalcemic and hypophosphatemic effects by inhibiting osteoclastic bone resorption and renal tubular phosphate reabsorption, respectively. CT receptors (CTRs) are members of a subfamily of seven-transmembrane domain, G protein coupled receptors that include those for several other peptide hormones. Basic amino acid substitutions within the CT molecule enhance potency, probably by conferring a helical structure to the peptide. This might explain the enhanced potency of fish CTs for mammalian CTRs. The presence and secretion of salmon CT like immunoreactive material have been described in both the murine and human central nervous systems, which possess CTRs. These findings are consistent with a role for this peptide acting as a neurotransmitter in mammals. Stanniocalcin (STC) is another hypocalcemic hormone originally identified in fish. In fish STC exerts its anti-hypercalcemic effect by regulating calcium and phosphate transports by the gills, intestine and kidney. Although fish ultimobranchial cells are much less responsive to the secretagogic effects of Ca2+ than mammalian parafollicular cells, the secretion of both CT and STC are positively regulated by extracellular calcium. STC has also been recently identified in humans and rats. It is released by some renal tubular cells and might play a role in the regulation of phosphate metabolism. Nevertheless, the true physiologic roles for CT in fish and STC in mammals, respectively, remain unknown. PMID- 9864548 TI - [Seventh congress of the National Federation of Endocrinology, Diabetes and Metabolism Regional Associations]. PMID- 9864549 TI - [New conditions for the use of growth hormone and outcomes for different indications]. PMID- 9864550 TI - [Somatostatin agonists in the treatment of acromegaly]. PMID- 9864551 TI - [Polycystic ovary syndrome: from physiopathology to therapy]. PMID- 9864552 TI - [Hyperandrogenism and fertility]. PMID- 9864553 TI - [Value and contribution of ovarian reserve markers]. PMID- 9864554 TI - [Cardiovascular risk and diabetes]. PMID- 9864555 TI - [Cardiovascular risk and dyslipidemias]. AB - The relationship between plasma lipid abnormalities and the risk for cardiovascular disease is now clearly established. In addition to quantitative abnormalities such as increased plasma LDL-cholesterol and decreased HDL cholesterol levels, lipid qualitative abnormalities are likely to play an important role in the pathogenesis of atherosclerosis. Indeed, dense triglyceride rich LDL particles are atherogenic. Moreover, oxidized LDL are associated with increased cardiovascular risk and may play a central role in the pathogenesis of the atherosclerotic plaque. Fasting triglyceride level has rarely been shown to be an independent risk factor for cardiovascular disease, in the general population, but plasma triglyceride levels, following an oral fat load, seem to be a better marker. Recent intervention studies have shown an early effect of HMGCoA-reductase inhibitors in reducing cardiovascular morbidity and mortality, suggesting possible additive actions of statins on the atherosclerotic plaque, on the endothelial function and on thrombosis. The Care study shows that a clinical benefit with pravastatin treatment, in secondary prevention, can be obtained in patients with LDL-cholesterol above 125 mg/dl. If all the primary and secondary intervention studies have shown reduction of cardiovascular morbidity and mortality with hypolipidemic drugs, we also have to keep in mind the positive results from the dietary intervention studies, showing beneficial actions of diets rich in vitamins, fibers and omega 3 fatty acids. In clinical practice, compliance of the patient to his treatment is often difficult to achieve and many studies have shown that control of hyperlipidemia is frequently inaccurate and needs to be reinforced. A comprehensive education of the hyperlipidemic patient is urgently needed in order to improve his knowledge about hypolipidemic diet and to obtain his motivation which is essential for a long term compliance. PMID- 9864556 TI - [Adrenal scintigraphy with 131I-6beta-iodomethyl-19-norcholesterol: good practice guideline]. AB - Adrenal scintigraphy with 131I-6 beta-iodomethyl-19-noncholesterol requires well prepared patients. Its interpretation requires trained observers with good knowledge of adrenal physiology and adrenal diseases. This multidisciplinary cooperative work was conducted by endocrinologists, nuclear medicine physicians and surgeons, in order to help physicians to optimize indications and practical implementation of this scintigraphy, which has constraints and pitfalls but is very informative. PMID- 9864557 TI - [Individualized cardiovascular prevention: when and how far should it go?]. AB - Individualised cardiovascular prevention must primarily be applied to patients with coronary heart disease or another obvious atherosclerotic disease and to apparently healthy subjects, but presenting a high risk (which implies the detection of these subjects) and their close relatives. The European recommendations for the prevention of coronary heart disease in clinical practice have been recently up-dated. Calculation of the multifactorial absolute risk (probability of developing an event over a given period, taking into account all modifiable and non-modifiable risk factors) represents a decisional aid to intervene on lifestyle and drug prescription. However, the available tables and software require certain adjustments. Diet is a major determinant of the cardiovascular risk and has one of the most important, but most complex impacts on prevention. Restriction of saturated fats, a Mediterranean type of diet and caloric restriction in the case of obesity are the main objectives. Smoking cessation is essential. For all these measures, a specialist referral is too frequently neglected. Aerobic physical activity is recommended. Hypertension must be controlled regardless of age. The objective is to obtain blood pressure figures less than 140/90 mmHg, and even lower in the case of diabetes or renal disease. Evidence of the benefit of controlling hypercholesterolaemia has been reinforced and the objectives to be achieved have been redefined: in secondary prevention and in high-risk subjects, the target total cholesterol must be less than 1.90 g/l and LDL cholesterol must be less than 1.15 g/l. The control of all risk factors must be reinforced in diabetic patients and fasting blood glucose must be maintained between 0.91 and 1.20 g/l in insulin-dependent diabetic patients with coronary heart disease. PMID- 9864558 TI - [Management of atherogenic hyperlipidemia]. AB - After defining the type of hyperlipoproteinaemia (the three main are IIa, IIb and IV), confirming its primary nature (i.e. after eliminating the various causes of secondary hyperlipoproteinaemia) and after defining the therapeutic objective (different according to the context of primary or secondary cardiovascular prevention), treatment must be prescribed. It always consists of diet, and very often a lipid-lowering drug. The remarkable results obtained with statins in studies of primary prevention, secondary prevention and regression of atherosclerosis does not mean that they can be prescribed blindly. Therapeutic indications depend on the type of hyperlipidaemia. By limiting the discussion to the three main types of atherogenic hyperlipoproteinaemia, treatment consists of: in pure type IIa hypercholesterolaemia: a first-line statin (or fibrate or resin in the case of adverse effects); in pure type IV hypertriglyceridaemia: a fibrate and possibly omega-3 fatty acids; in combined or mixed type IIb hyperlipidaemia: a statin in the case of type IIb with predominant hypercholesterolaemia, a fibrate in the case of type IIb with predominant hypertriglyce-ridaemia, failure to comply with these rules can lead to poor laboratory results. PMID- 9864559 TI - [The traps of pulmonary embolism]. AB - The clinical diagnosis of pulmonary embolism (PE), even massive, remains difficult and perplexing. In our hospital, the percentage of exact clinical diagnoses has not significantly changed over recent years, with a false-negative rate of 78%, and a false-positive rate of 2%. In approximately 20% of cases, autopsy showed several emboli and pulmonary infarctions of various ages, indicating recurrent embolic episodes. The diseases most frequently associated were cardiac diseases (51%) and tumours (24%). Pneumonia considerably decreases the probability of an exact diagnosis of PE; hospitalisation in the Cardiology department or Intensive Care Unit increased the probability of this diagnosis. While the numerous diagnostic algorithms recently proposed have a limited value, the process integrating clinical and instrumental data in order to establish a prospective clinical probability, should facilitate identification of acute PE in live patients. PMID- 9864560 TI - [Clinical application of the genetic aspects of familial hypertrophic cardiomyopathy]. AB - Familial hypertrophic cardiomyopathy is a disease with very heterogeneous genetic characteristics, both at the intergenic level (7 genes have been identified and some of them are still unknown) and intragenic level (more than 100 mutations have been identified). This genetic heterogeneity at least partly explains the clinical heterogeneity, expressed in terms of the degree of left ventricular hypertrophy and the inherent risk related to the disease (sudden death and/or heart failure). However, the understanding of phenotype/genotype relationships of this disease is only in the very early stages. So-called "modifier" genes are probably involved to modify the clinical profile of the disease. Many healthy carriers have been identified, but their outcome is unknown at the present time. The molecular genetics of HCM allows reevaluation of the classical clinical criteria. In the future, it would be desirable to establish specialized units for predictive and antenatal diagnosis, composed of cardiologists specialized in the disease, geneticists and psychologists in order to consider all clinical, ethical, and psychological aspects of this disease. Hopefully, this new approach to cardiology will be able to establish a more accurate risk profile for exposed subjects and guide treatment options. PMID- 9864561 TI - [Quantitative indices of mitral valve regurgitation]. AB - The assessment of the degree of mitral regurgitation has always been difficult and subject to errors. With the growing impetus on surgery in asymptomatic patients, the quantitation of mitral regurgitation has become a crucial part of the assessment of these patients. Several new methods have been developed that are currently utilized and allow the measurement of the lesion severity (Effective regurgitant orifice) and the volume overload (regurgitant volume and fraction) of mitral regurgitation. PMID- 9864562 TI - [Quantification of chronic aortic insufficiency by transthoracic Doppler echocardiography]. PMID- 9864563 TI - [Qualitative approach to aortic valve insufficiency: role of echography in the therapeutic indication and choice of surgical method]. PMID- 9864564 TI - [The role of preoperative echography in the surgical reconstruction of the mitral valve]. PMID- 9864565 TI - [Stress echography]. AB - Although stress echocardiography was introduced in the beginning of the 1980s, it has only been widely used since the appearance of digital technology. This technique has undergone rapid development for the evaluation of coronary artery disease because of its diagnostic and prognostic contribution. PMID- 9864566 TI - [Doppler imaging of myocardial tissue: useful in research of aid to the clinician?]. PMID- 9864567 TI - [Contrast echocardiography and myocardial perfusion: what is the current status?]. AB - Progress in the field of echocardiographic contrast agent combined with progress in imaging techniques (second harmonic imaging, intermittent imaging, Doppler Energy) should allow a real revolution in the field of noninvasive cardiac imaging, and one of the main advantages will probably be myocardial perfusion imaging in ischaemic heart disease. PMID- 9864568 TI - [Constriction or restriction: does echocardiography indicate the difference?]. PMID- 9864569 TI - [Does transesophageal echocardiography modify the managements of atrial fibrillation?]. PMID- 9864570 TI - [Atheroma of the aortic arch and embolic risk]. AB - Aortic atheroma, involving the ascending aorta and proximal aortic arch, is associated with an increased risk of cerebral and peripheral embolic complications and cardiovascular complications (acute myocardial infarction, vascular death, peripheral arterial ischaemia). These data were established on prospective, case-control, anatomical studies, which all used transoesophageal echocardiography as the reference diagnostic method. In the absence of an alternative radiological imaging technique, transoesophageal echocardiography describes the thickness of the plaque, its luminal extension, its more or less regular nature, and the presence of plaque calcifications and sessile or mobile thrombi. Some predisposing factors are associated with the thromboembolic risk of aortic athero-sclerosis: elevated plasma fibrinogen, presence of circulating anticoagulants, elevated plasma homocysteine. The mana-gement of these patients is purely empirical at the present time. Antiplatelet treatment is formally indicated following cerebral infarction. The presence of thrombosis on the atherosclerotic plaque constitutes an indication for anticoagulant therapy. The indication for thrombolysis and surgical resection of the aortic atheroma is anecdotal, in the absence of proof of their efficacy on large series. Progress in radiological and echocardiographic imaging, and complementary epidemiological studies should be able to more clearly define patient subgroups at higher risk. Aortic atherothrombosis was also recently associated with increased cardiovascular complications in patients with atrial fibrillation (SPAF III trial). PMID- 9864571 TI - [Cancer of the colon: the role of the pathologist]. PMID- 9864572 TI - [Telomerase and cancer: importance of detection of telomerase activity in pathology]. PMID- 9864573 TI - [Application of the PRINS technique for chromosome examination in fetal cells present in maternal blood]. AB - Both detection and chromosomal analysis of fetal cells present in the maternal circulation can be performed using Histopaque double density gradient centrifugation followed by primed in situ (PRINS) labeling technique. This approach has been tested on blood samples from 15 pregnant women and 6 control donors with primers specific for chromosomes 9, X and Y. The cell separation technique allows recovery of both mononuclear cells and polynuclear cells with a 97% efficiency. PRINS labeling was successful in 100% cells from control blood samples. Among patient samples, 2 "false-negative" results were observed. These preliminary results suggest that the present protocol might be efficient for non invasive prenatal chromosome analysis. PMID- 9864574 TI - [Reproducibility of histoprognostic grades of invasive breast cancer]. AB - In the framework of the quality assurance of the pathologists involved in the screening campaign it was proposed to test the reproducibility of the classical SCARFF, BLOOM and RICHARDSON (SBR) grade and the ELSTON-ELLIS grade (E-E) mentioned in the recent European recommendations. Twenty-four cases of invasive carcinoma were analyzed by 21 pathologists. Sixty-eight per cent agreement was obtained for the SBR grade and 69% for the E-E grade in the first round and vice versa in the second round. The mean value of the kappa test was 0.54 with SBR and 0.53 with E-E. The differentiation of the carcinoma was found to be the most reliable index (kappa score greater than 0.5), followed by the mitotic index (kappa score = 0.39 for SBR and 0.36 for E-E), and the nuclear polymorphism obtained the lowest score (kappa score ranging from 0.33 to 0.38). In our hands, the use of the E-E grade did not improve the reproducibility particularly for the mitotic index. Our results suggest however that the reproducibility may be improved by training. PMID- 9864575 TI - [Telomerase and cancer]. PMID- 9864576 TI - [Which histo-prognostic factors are useful for a therapeutic decision in colon cancer?]. PMID- 9864577 TI - [Hyperplastic polyp in heterotopic gastric mucosa. A rare lesion of the cervical esophagus]. AB - Patches of heterotopic gastric mucosa are often endoscopically detected in the upper third of the esophagus. A hyperplastic polyp developing on one of those heterotopic fundic mucous areas is a rare event. In this paper, a case revealed by pyrosis is reported in a 60-year-old man. Though there may be malignant potential, no malignant transformation of such a lesion has been reported so far in the esophagus. PMID- 9864578 TI - [The intratesticular leiomyoma: an unusual location. Report of a case]. AB - Intratesticular location of leiomyoma is unusual. A single case has been published in the literature. We report a case of what we consider to be an intratesticular leiomyoma, with a description of its pathology, a discussion of its differential diagnosis and histogenesis. PMID- 9864579 TI - [Kikuchi-Fujimoto subacute necrotizing lymphadenitis: two histologic forms observed in the same patient]. AB - Histiocytic necrotizing lymphadenitis, also known as Kikuchi-Fujimoto disease is a rare self-limited condition of young adults usually misdiagnosed as malignant hematologic disease. The diagnosis depends on microscopic findings. Two histologic types are classically described, proliferative and necrotic types. This is a case report of 17 year-old girl who presented cervical lymphadenopathy. The first lymph node biopsy was in favor of malignant lymphoma. The second lymph node biopsy, done one month later, showed typical histiocytic necrotizing lymphadenitis. To our knowledge, this is the first time that the two forms of this disease are described in the same patient. PMID- 9864580 TI - [Arrhythmogenic right ventricular cardiomyopathy. A pathologic diagnosis to invoke in the sudden death of a young subject]. AB - We report a case of arrhythmogenic right ventricular cardiomyopathy revealed by sudden death during exercise in a 13-year-old patient. Postmortem diagnosis was made on multiple tissue samples taken from right ventricular free wall, showing light adipous infiltration of the myocardium at gross examination. Arrhythmogenic right ventricular cardiomyopathy is histologically characterized by fibro-fatty replacement of right ventricular myocardium. Left ventricular involvement may be observed. Diagnosis at an early stage is often difficult. Etiology remains unknown. Since familial occurrence has been documented, postmortem identification is useful for the other members of the family. PMID- 9864582 TI - [A peculiar tonsillitis...]. PMID- 9864581 TI - [Castleman's disease and chromophobe carcinoma of the kidney. An incidental association?]. AB - We report a case of Castleman's disease in a 65-year-old female, revealed by a renal tumor associated with inter-aortico-cava adenopathies and renal chromophobe cell carcinoma. This observation points to the difficulties in differentiating local Castleman's disease, which may be cured by surgical excision, from multicentric disease associated with a dysimmune syndrome of uncertain prognosis. The association of multicentric Castleman's disease with a carcinoma has rarely been described. It could be the emergence of a neoplasia in a context of dysimmunity or Castleman's disease might be related to the production of interleukin 6 by a renal carcinoma. PMID- 9864583 TI - [A bladder tumor which should not go undiagnosed]. PMID- 9864584 TI - [An uncommon parotid tumor]. PMID- 9864585 TI - [Diagnosis and prognosis of malignant colonic adenoma: methodologic implications]. AB - The subsequent management of patients with endoscopically removed malignant colorectal polyps depends on the pathologist's diagnosis. According to the correlation with clinical outcome previously published, the histological criteria of potential adverse outcome are: the presence of tumour at or near the resection margins, the invasion of colonic submucosa, the lymphatic invasion and the high grade of malignancy. This article aims at providing guidelines for an adequate examination of the polyp specimen and an unambiguous pathology reporting. PMID- 9864586 TI - [Recommendations for editing pathology reports of colorectal cancer]. AB - The purpose of these recommendations is to provide an informative report for the clinician. The recommendations have been divided into two major areas 1) a standardized form, 2) items that provide an informative gross description and diagnostic features that are recommended to be included in every report. In special circumstances, the recommendations may not be applicable. PMID- 9864587 TI - [Ki-67 and apoptosis in a embryologic model]. PMID- 9864589 TI - Monitoring for intraoperative awareness. AB - Despite advances in anesthetic technique and technology, intraoperative awareness continues to occur with alarming regularity. The psychological effects on patients and the medicolegal consequences to providers have created the need for a reliable monitor of the hypnotic component of anesthesia. A new technology known as bispectral analysis (BIS) has the potential to put an end to the devastating occurrence of intraoperative awareness, as well as improve the overall effectiveness of anesthetic delivery and the surgical experience. Applicable to almost any situation requiring sedation or general anesthesia, BIS's reliability is a real-time measure of hypnosis levels. PMID- 9864590 TI - Renal transplantation. AB - Medical and scientific advances have improved the diagnosis and treatment of kidney disorders. Organ transplantation has evolved from an experimental surgery to a medically accepted form of treatment for organ failure. The kidney was the first organ to be successfully replaced by a donor organ, and it is presently the most commonly transplanted organ. Kidney transplantation restores reasonably normal health to patients whose kidneys no longer function, and it frees them from the limitations imposed by dialysis. Improved graft survival rates have further enhanced the desirability of transplantation. PMID- 9864591 TI - The effects of local anesthetics on postoperative pain. AB - This study was performed to determine if intraoperative local anesthesia improved control of postoperative pain after inguinal herniorrhaphy and to compare the effects of two commonly used local anesthetics on pain management. The Gate Control Theory of Pain formed the theoretical basis for this study. A retrospective nonexperimental study in an ex post facto design was used. Data were collected from 1990 through 1997 on 120 patient charts. The use of local anesthetic intraoperatively significantly decreased patients' lengths of stay postoperatively (P = 0.00) and need for postoperative narcotics (P = 0.00). Bupivacaine was found to be superior to lidocaine in decreasing the need for postoperative narcotic analgesia. Researchers concluded that many patients would benefit from intraoperative injection of local anesthesia. This information can affect patient care outcomes through decreasing recovery time, reducing postoperative pain, and reducing health care costs. PMID- 9864592 TI - Nurses find solution to postoperative leg and back pain. PMID- 9864593 TI - Transplant information on the World Wide Web. PMID- 9864594 TI - Dispelling the myth that research answers all questions. PMID- 9864595 TI - Recommended practices for surgical attire. Association of Operating Room Nurses. PMID- 9864596 TI - Recommended practices for use of the pneumatic tourniquet. Association of Operating Room Nurses. PMID- 9864597 TI - [Heart failure management: a challenge for the 3rd millennium]. PMID- 9864598 TI - [Epidemiology and economic impact of heart failure in France]. AB - There are little epidemiological data available about heart failure in France, despite its considerable impact on the system of health care and the fact that this problem will become even more acute in the future. Here are some important statistics: in France, there are about 500,000 people suffering from heart failure with about 120,000 new cases every year. The incidence increases from 4% in men and 3% in women of 55 to 64 years of age to 50% in men and 85% in women of 85 to 94 years of age. The average age of diagnosis is 73.5 years: two thirds of patients are over 70 years of age. There are about 3.5 million consultations and 150,000 hospital admissions for heart failure per year. The average length of hospital stay is 11 days. There are more than 32,000 deaths per year from heart failure. The cost of treating heart failure represents more than 1% of total medical expenses. Heart failure is a major problem of public health which is on the increase. This should incite physicians to provide optimal treatment for those affected and to place greater emphasis on preventive measures. PMID- 9864600 TI - [Can the results of large-scale clinical trials be applied to clinical practice?]. AB - The management of a patient in cardiac failure should lead the physician to ask if the patient might benefit from treatment with an angiotensin converting inhibitor. Meta-analyses give a positive reply but often lack detail. Analysis of large scale therapeutic trials is the only way to determine whether the results are applicable in an individual case. This analysis starts with validation of the methodology (correct randomisation, respect of double blind protocol, identity of groups) and synthesis of results expressed, if possible, in absolute values. In a second stage, the criteria of inclusion, exclusion, and the comorbidity of enrolled patients, should be compared with those of the patient under consideration. In addition, the amplitude of the therapeutic benefit and the importance of undesired effects must be taken into account in the therapeutic decision. This approach allows a reasoned prescription based on documented facts. PMID- 9864599 TI - [Physiopathological basis of the treatment of heart failure]. AB - Therapeutic advances have changed the mode of presentation of cardiac failure over the last decades: the main cause, nowadays, is myocardial ischaemia. The modern treatment of cardiac failure is based on relatively simple physiopathological mechanisms which take into account the different aspects of cardiac physiology: a pump, a muscle, a coronary circulation supplying oxygen to the myocardium, an automatic contraction. The concept of vasodilatation and the blocking of vasoconstrictive systems introduced during the 70s is the basis of modern treatment of cardiac failure which involves angiotensin converting enzyme inhibitors and, increasingly, betablockers. In the near future, with earlier treatment of cardiac failure, the stimulation of vasodilator systems could become a new therapeutic strategy. Early detection of ischaemia and its complications with the aim of limiting the loss of cardiac myocytes is a priority for slowing the progression of cardiac failure. The prevention of cardiac failure also depends on educating cardiologists to treat rapidly the factors predisposing to or prolonging episodes of even mild cardiac failure. PMID- 9864601 TI - [Prevention of heart failure]. AB - Heart failure is a major problem of public health, associated with poor outcome in the advanced stage, thus justifying its prevention. Primary prevention is based on the prevention and treatment of its principal etiologic factors, hypertension and coronary artery disease. Broad use of echocardiography or dosage of neurohormonal markers improve detection of asymptomatic left ventricular dysfunction. In ischemic heart disease, coronary recanalisation prevents or limits left ventricular remodeling and dysfunction, even if the "open artery" theory has not been entirely proved. Understanding the deleterous role of neurohormonal stimulation results in a large use of ACE-inhibitors, which beneficial effect has been demonstrated also in case of asymptomatic left ventricular dysfunction. Betablockers, already largely used after myocardial infarction, seem to have also a beneficial effect in heart failure: the same is probably also true for angiotensin II-antagonists. Double blocking of both the sympathetic nervous system and the angiotensin-aldosterone system seems to be recommended. More precisely understanding the pathways signaling the processes of ventricular remodeling and dysfunction points to new potential targets for a preventive treatment: endothelin receptors, apoptosis, oxidative stress, cytokines or even angiogenesis. PMID- 9864602 TI - [Management of patients with symptoms on effort]. AB - The aim of the treatment of symptomatic cardiac failure on effort (Classes II and III) is to improve symptoms and functional capacity and to decrease mortality. The management of such patients includes the investigation of causal and/or aggravating factors which require specific treatment. Hygenic-dietetic rules are important with a low salt diet and restriction of water intake. The continuation of mild physical exercise should be encouraged. Medical treatment associates diuretics to reduce congestive symptoms and angiotensin converting enzyme inhibitors to improve survival. This treatment may be completed by the association of digoxin and/or carvedilol depending on the functional outcome. When stabilised, the patient's diuretic should be reduced to the minimal required dose. If patients remain symptomatic on quadruple therapy, they probably require reevaluation in hospital. PMID- 9864603 TI - [Therapeutic strategy of the management of acute heart failure]. AB - Acute cardiac failure should be approached clinically and paraclinically as an emergency and precipitation is not always what is required. Consequently, strict clinical and paraclinical strategy leads to decisive action to improve the prognosis. The authors describe the principles of management of acute cardiac failure. The different stages which are carried out simultaneously in clinical practice are discussed: physiopathological basis, clinical and paraclinical investigations, symptomatic and aetiological treatment. The description and indications of circulatory assistance are considered. This constitutes a major therapeutic advance of the last decade and has changed the treatment of the most severe clinical forms of acute cardiac failure. PMID- 9864604 TI - [Is the treatment of left ventricular systolic dysfunction different according to the etiology?]. AB - Cardiac failure is the terminal stage of evolution, the finality of many valvular, vascular, myocardial, general, congenital or acquired conditions. The therapeutic decisions should be based on the search for a curable cause of a predisposing factor and the evaluation of the severity of the cardiac failure. At advanced stages of ventricular dysfunction when the myocardial lesions are constituted, when cardiac and vascular remodelling has occurred, the aetiological treatment, which is the constant objective, is unfortunately too late. The treatment is the same, whatever the aetiology, in order to improve functional problems. At early stages, and, if possible, preventively, surgery, revascularisation techniques, the correction of an arrhythmia, the suppression of a cardiotoxic factor, are essential. The different therapeutic classes used could have different efficacies depending on the aetiology, but, finally, this point is negligible: the medications are based on the results of large scale, controlled, therapeutic trials. PMID- 9864605 TI - [Diastolic cardiac failure: therapeutic modalities]. AB - The management of cardiac failure due to diastolic dysfunction is not well codified and is often empirical. It has three objectives: improving the physiopathological components of ventricular filling, treating the associated aggravating pathological conditions, and treating the basic cause of the dysfunction. Symptomatic treatment aims to reduce venous congestion (by diuretics or nitrate derivatives), to prolong the diastolic period by slowing the heart rate (by betablockers, bradycardising calcium antagonists or digitalis in cases of irreducible atrial fibrillation), to improve passive ventricular distensibility by an effect on remodelling (by angiotensin converting enzyme inhibitors or anti-aldosterone diuretics). The treatment of associated pathological conditions is particularly important. It is essential to maintain or reestablish an effective atrial systole by cardioversion and anti-arrhythmic drugs in atrial fibrillation, by dual chamber pacing in cases of atrioventricular asynchrony due to atrioventricular block. Treatment of the underlying cause aims to induce regression of ventricular hypertrophy of hypertensive origin by using antihypertensive drugs with this property. In coronary artery disease, the choice is determined by the clinical context because nearly all anti-anginal or interventional treatments may improve ischaemic diastolic dysfunction. The same applies in hypertrophic cardiomyopathy because most types of treatment (betablockers, verapamil, cardiac pacing, surgery) may improve diastolic function. Finally, in valvular aortic stenosis, aortic valve replacement restores normal diastolic function. PMID- 9864606 TI - [Management of arrhythmias in patients with heart failure]. AB - Cardiac failure is a common cause of arrhythmia. Many factors predispose to the genesis of arrhythmias in these patients. A number of non-invasive methods allow stratification of the risk of arrhythmia in cardiac failure. Approximately half the deaths of these patients are due to arrhythmia. Unfortunately, most of the investigations for risk evaluation have a high negative predictive value but a lower positive predictive value. The treatment of supraventricular arrhythmias, mainly atrial fibrillation, is complex in cardiac failure. Class I antiarrhythmics are contraindicated. The only remaining options are Class II, especially Sotalol, and Class III drugs, especially Amiodarone. In some cases, non-pharmacological methods such as ablation, pacing or an implantable atrial defibrillator must be considered. The treatment of ventricular arrhythmias is also difficult. In this indication, Class I antiarrhythmic agents must also be avoided. Non-sustained ventricular tachycardia may be treated by betablockers or amiodarone. The use of an implantable defibrillator is increasingly recommended after the results of several controlled large scale trials. The indication is obvious in patients resuscitated from sudden death and these devices are also beneficial in sustained ventricular tachycardia in patients with cardiac failure. Many studies are currently under way to determine the value of this therapeutic modality in indications now considered to be "prophylactic". PMID- 9864607 TI - [Aspirin, oral anticoagulants, and heart failure]. AB - The incidence of thromboembolic complications in patients with cardiac failure is low. The predisposing factors are principally the presence of a left ventricular mural thrombus, atrial fibrillation, a low ejection fraction and a low peak VO2. The risk of cerebral haemorrhage in a patient with cardiac failure treated with oral anticoagulants is about the same as the risk of thromboembolism. Therefore, anticoagulant therapy for patients with cardiac failure is controversial in the absence of a prospective large scale clinical trial demonstrating its benefits. In the meantime, a prudent approach with risk stratification to determine which patients would benefit the most from oral anticoagulation is advised. PMID- 9864608 TI - [Diuretic resistance: mechanisms and therapeutic possibilities]. AB - The diuretics, with the exception of spironolactone, act on the luminal (or apical) surface of the tubular cells of different segments of the nephron. In order to act, they must be secreted into the tubular lumen. This transfer of the drug to its site of action may be blocked by decreased renal blood flow, the saturation of the systems of tubular transport or fixation to the albumin present in the primary urine. All these pharmacokinetic abnormalities (observed in renal failure or the nephrotic syndrome) lead to diuretic resistance. Increasing the dosage, the repetition, intravenous administration, even as an infusion, are possible solutions. Resistance may be observed in the absence of pharmacokinetic abnormalities: in these cases, there is an abnormal response of the tubular cells to otherwise effective diuretic concentrations, or the activation of homeostatic mechanisms leading to sodium retention and preventing negativisation of the salt and water balance. These situations are often associated in cardiac failure or cirrhosis with oedema. Increasing the dosage is not a logical solution, but increasing the frequency of administration may be helpful. The importance of secondary hyperaldosteronism in cirrhotic oedema makes spironolactone the treatment of choice. In all cases, the addition of two mechanisms of inhibition of tubular reabsorption of sodium at different sites in the nephron often results in an effective diuresis: usually, this implies the addition of a thiazide (e.g. hydrochlorothiazide) to an initial prescription of a loop diuretic. PMID- 9864609 TI - [Pacing and heart failure]. AB - Conventional dual-chamber cardiac pacing with the right ventricular pacing catheter positioned at the apex of the right ventricle was proposed eight years ago by Hochleitner as primary treatment of refractory cardiac failure. Although the preliminary results were encouraging, those at long-term and subsequent studies were less favourable in this indication. Biventricular pacing is now a technical possibility and allows at least partial correction of the electromechanical abnormalities observed in dilated cardiomyopathy. The preliminary results of biventricular pacing in patients with refractory cardiac failure due to dilated cardiomyopathy with major intraventricular conduction defects are very encouraging. This new therapeutic approach should now be validated by controlled, randomised therapeutic trials. PMID- 9864610 TI - [Rehabilitation by physical exercise in chronic heart failure]. AB - The aim of rehabilitation is to improve exercise capacity and, thereby, the autonomy of patients with cardiac failure. For many years, these patients were considered inapt to perform physical exercise and they are in the same situation at the dawn of the year 2000 as patients with myocardial infarction forty years ago. The symptoms of cardiac failure (dyspnoea of effort and muscular fatigue) are not only the consequence of pulmonary hypertension and decreased muscular perfusion. Prolonged interruption of exercise and long stays in bed or in a chair lead to anatomical and functional amyotrophy, which, in turns, incites to further inactivity. Deconditioned respiratory muscles cannot tolerate the increased load of hyperventilation. Neurohormonal changes cause vasoconstriction which reduces muscular perfusion. Physical training can significantly improve these abnormalities, though it does not seem to have a measurable effect on cardiac function; based on segmental work which enables performance of substantial efforts with a minimum of haemodynamic changes, it provides a 20 to 30% gain in capacity, mainly increasing the duration of submaximal exercise rather than maximum performance. Muscular fatigue is the symptom which is the most improved. Unfortunately the organisation, which is more difficult than in the post infarction period, and the generalisation of the practice of long-term, well adapted physical training remains marginal although hundreds of thousands of patients could benefit; more than the inertia of the official instances concerning anything related to cardiac rehabilitation, it is the lack of interest shown by cardiologists and the absence of flexible structures within the health care organisation for elderly people which are responsible. PMID- 9864611 TI - [Education of patients with heart failure]. AB - Chronic cardiac failure is a common complication of coronary artery disease and hypertension. It is observed above all in the elderly and represents a high public health cost, mainly due to hospital admissions. The education of patients with cardiac failure, and their families', will contribute to an improved cost/efficacy ratio of the management of this pathology. This educative process comprises a simple description of cardiac failure, its symptoms and aetiologies. Dietary advice is very important (salt intake, drinks...). Physical exercise, which is not contraindicated, should be adapted to the degree of functional impairment and the same applies to travelling and voyages. Medical treatment should be clearly explained, mode of action, dosage, expected benefit, side effects... Finally, the possible progression of the disease should be explained if arrangements have to be made. The possibility of cardiac transplantation should be discussed if the subject is young. Although the risk of sudden death is not usually spoken about, this common complication of cardiac failure justifies consideration and widespread instruction of cardiopulmonary ressuscitation. PMID- 9864612 TI - [Alternative surgical options to heart transplantation]. AB - Cardiac transplantation is the treatment of reference for refractory cardiac failure but the limited number of donors, the complications inherent to transplantation and the relative and absolute contra-indications has made it necessary to find alternative surgical solutions. The detection of myocardial viability by Thallium scintigraphy, Dobutamine echocardiography and/or position emission tomography in coronary disease, allows identification of zones which are capable of recovering contractile function after revascularisation. The authors report the results of a series of 91 operated patients with a 10 year follow-up having a 72% 5 year actuarial survival and improved ejection fraction. The other alternative which may improve symptoms and prognosis in patients with severe ischaemic heart disease with left ventricular dysfunction is apical remodelling or Dor's procedure. The results of a haemodynamic study at 1 year of 171 patients clearly show a functional improvement and an increase of the ejection fraction. The advantage of this method is that it can be used in patients with dyskinetic and akinetic plaques resulting from antero-septo-apical infarction. Finally, even if mitral regurgitation is relatively uncommon in chronic ischaemic heart disease, a simple procedure (annuloplasty) is often sufficient to correct the mitral regurgitation and reduce the afterload of a failing ventricle. On the other hand, in dilated cardiomyopathy, two new options have been developed; one, suggested by Steven Bolling, proposes simple mitral annuloplasty whatever the underlying cause (primary or ischaemic cardiomyopathy) with symptomatic improvement and better haemodynamics in terms of increased cardiac output and oxygen consumption on exercise and an actuarial survival much higher than that of cardiac transplantation at one and at two years. The most recent innovation is the Batista procedure which is a method of ventricular reduction associated with correction of mitral regurgitation. The authors have assessed 20 patients for this operation at the Foch Hospital by Dobutamine echocardiography and 5 patients underwent the procedure. All 5 patients reported symptomatic improvement but some had an unchanged haemodynamic status. Others improved at rest and some improved on exercise. The Cleveland Clinic series reported results in 57 cases. Whichever alternative method tested, there is a significant functional improvement but the cardiac output does not always increase. There are no comparative prospective randomised studies and strict selection of patients is required, a problem not yet resolved for all indications. The advantages of these procedures are certain as there is no waiting list, the functional results in good indications have been demonstrated and, if necessary, secondary orthotopic cardiac transplantation is always possible. PMID- 9864613 TI - [Drugs of the future]. AB - Many therapeutic approaches are under evaluation in patients with cardiac failure. They include angiotensin receptor inhibitors, selective and non selective endothelin receptor inhibitors, neutral endopeptidase inhibitors or mixed inhibitors of neutral endopeptidase and of the angiotensin converting enzyme and, finally, cytokinin modulators. Some of these drugs have already entered Phase II therapeutic trials and are at relatively advanced developmental stages. Others are at preliminary or experimental stages. If these new drugs prove to be effective and well tolerated, they will represent new tools for physicians to treat cardiac failure and prevent its progression. However, many questions concerning drug associations and poly-therapy will be raised, leading to a revision of the strategy of treatment of cardiac failure. PMID- 9864614 TI - [Cellular cardiomyoplasty: state of the art, evaluation, and future possibilities]. AB - Cellular cardiomyoplasty, or the transplantation of myogenic cells into the myocardial tissues, could emerge as a therapeutic alternative in patients with cardiac failure. It depends on several procedures: implantation of cell types, syngenic embryonic cardiomycocytes, allogenic and autogenic cardiac muscle cells. These cells carne into contact with host cardiomyocytes and could contract in a synchronous fashion. Experimental data suggests that this technique could improve global left ventricular function in the post-infarction period or in dilated cardiomyopathy even though the precise mechanism of this improvement is not fully understood. Many difficulties remain, the cell types have an oncogenic potential; syngenic foetal cells are weakly immunogenic but their use is limited by ethical and problems of supply. Therefore, auto-transplantation either of cardiomycocytes obtained by endomyocardial biopsy or of adult skeletal muscle, could be a potential clinical option. PMID- 9864615 TI - [Telecommunication in trauma surgery. Communication networks of hospitals in East Bavaria]. AB - The growing complexity of the performance processes in medicine makes it mandatory that the flow of information is faster and more consistent, especially when the sites of health care are far away from each other. The Regensburg model, a realization of lean telemedicine from a low-cost domain, using PC-based standard videoconferencing systems shows the use of modern telecommunications, especially in trauma surgery. In 203 prospectively evaluated teleconsultations between 15 participants a total of 697 images were transmitted via videoconferencing. In 95% of the trauma cases the transmitted material was judged as at least sufficient. In project-attending evaluations the efficacy of these systems and their use were clearly demonstrated. Savings in transportation costs of up to 4,400 DM per case were achieved. Through quicker flow of information quality improvements for all participants resulted; to some extent considerable costs for health care were avoided or reduced. Based on these thoughts, a new platform of communication will be established in Regensburg as a closed medical intranet for the region of eastern Bavaria. PMID- 9864616 TI - [Teleconsultation]. AB - Teleconsultation is a consultation between two or more physicians about the diagnostic work-up and therapeutic strategy in the treatment of an individual case by means of modern telematics. Due to more complex therapeutic strategies and legally defined formal requirements, the need for teleconsultation will increase significantly in the future. Rapid technical improvements in telematics will progressively facilitate the practical performance of teleconsultation (based upon an ISDN network in the beginning, later on by the use of a national health network). The medicolegal aspects of teleconsultation have already been defined sufficiently for use in surgery. However, the question of adequate financial compensation for this type of medical service is still unclear. PMID- 9864617 TI - [Teleconference and telesurgery]. AB - Interchange of information has become possible independent of the user's place or time by modern telemedical services and opens up new dimensions for diagnostic and therapeutic procedures. For teleconferences and teleconsultations in surgery, the intraoperative high-quality transmission of live images is essential without disturbing the sterility and operation routine. The clinical specifications require special systems for broadband, stereoscopic online image transmission both intra- and inter-institutionally. Moreover, a telemedical concept integrates the communication equipment suitable for the different diagnostic and functional facilities of a clinic, a digital multimedia patient record and modules for cooperative working. These implementations are a prerequisite for extensive telesurgical interventions using navigational tools, guided instruments or autonomous roboter systems. The realization of such a comprehensive telesurgical concept (OP 2000) is presented. PMID- 9864618 TI - [Analysis of leg geometry--standard techniques and normal values]. AB - The diagnosis of malalignments of the lower extremities includes analysis of the geometry of the whole leg. The first step in the diagnostic process is a standardized physical examination. It provides valuable background information for an effective radiological diagnosis. Even with a thorough standardized physical examination it is not possible to define exactly the deformity or decide on an operative procedure. The diagnosis of axis deviations in the frontal plane can be measured on a conventional plain X-ray of the whole leg. In this view it is very important that the knee joints are in a true a.p. view independent on torsional deformities of the lower legs. Today the gold standard to measure the torsion and length of the lower extremities is the CT scan. However, the multitude of analytical methods for CT measurements described in the literature do not lend themselves readily to comparison; thus, it is difficult to identify a clear method of choice. Not every CT measurement is better than a physical examination. Evidence of reproducibility and accuracy is a prerequisite for useful interpretation of the results. Up to this point in the literature there are only reference values for the Ulm CT Method. One alternative is the MR scan, which avoids radiological risks, but the reproducibility and accuracy of the MRI method are not as good as for the CT method. Another alternative is ultrasound, where recent advances in the measurement of torsion and length of the lower extremities have proven competitive with or superior to the accuracy of MRI. The three-dimensional determination of the torsion and length of the lower extremities by ultrasound has now assumed a leading role in the non-radiological diagnosis of malalignments of the lower extremities in children and adolescents. This method furthermore is increasingly being used in preoperative planning of leg deformities in adults. PMID- 9864620 TI - [Single corrective osteotomy after knee para-articular fractures]. AB - Post-traumatic malalignment can occur on one or several planes following fractures in close proximity to the knee joint. In a clinical and radiological analysis of deformities, the frontal, sagittal and longitudinal alignment, as well as differences in femoral or tibial length and torsion, must be taken into account. The location of the correctional osteotomy is usually defined by the center of the angular and torsional deformity. Of decisive importance when choosing the site of the osteotomy are the local quality of bone and the soft tissue envelope, the condition of the articular cartilage in the different compartments of the knee, the stability of ligamentocapsular structures and preexistent deformities of the distal femur and proximal tibia. Therefore, the location of correctional osteotomies must be modified, depending on the individual situation. The appropriate approach and technique must be chosen from the different available operative techniques. Inaccurate planning and inappropriate operative techniques pose severe risks. Specific postoperative complications are compartment syndromes, nerve irritation and infection. PMID- 9864619 TI - [Osteotomy of the proximal femur for correction of post-traumatic changes]. AB - Basically, four post-traumatic conditions of the proximal femur can be improved by corrective osteotomies: recurring luxations and subluxations of the hip joint; necrosis of the femoral head; non-unions; deformities. Dependent on the individual situation, the following procedure can be recommended, including diagnosis of deformities and indication and therapy of corrective osteotomies: analysis of the problem from the viewpoint of history, clinical findings and imaging techniques; indication for corrective osteotomy; graphic planning; operation technique. Particularities of post-traumatic conditions with regard to diagnosis, indication and surgical technique are indicated. Only with perfectly tailored concepts can we respond to the individual situation of an individual patient and reflect the variety and complexity of post-traumatic conditions of the proximal femur. PMID- 9864621 TI - [Correction of post-traumatic diaphyseal malalignment of the lower extremity]. AB - Post-traumatic deformities do occur as a result of disturbed fracture healing with loss of bone stock, necrosis of fragments or the development of pseudarthrosis resulting in malunion or progressive malalignment. In the majority of the cases these disturbances can be related to technical problems of primary fracture treatment like insufficient reduction or implant failure. On the other hand, complex deformities with involvement of the adjacent joints may also be a result of injuries of the growth plate in childhood. In some cases primary correction is impossible because of critical conditions of the bone and the soft tissue envelope in the center of deformation. If a secondary correction is indicated, knowledge of all reconstructive techniques is essential to choose the appropriate method and carry out successful and exact correction of malalignment of the affected limb after detailed planning. PMID- 9864623 TI - [Indications and technique of bone cutting]. AB - For cutting bones different tools and techniques are available. The question which method is most advantageous depends on the basic surgical concept, the location of the bone cut and the choice of the subsequent implant. The biological activity of the anatomical site of the cut determines how much the tissue needs to be protected, taking into consideration the vascularity of the bone and avoiding heat transfer. The surgical approach depends on the implant and should be used also for the bone cut. Specific indications are given for the oscillating saw, the Gigli saw, the osteotome, drill holes and the medullary saw. PMID- 9864622 TI - [Supramalleolar corrective osteotomy]. AB - The general aspects for the analysis of malalignment of the low tibial region in the three-dimensional space are discussed. Recommendations of clinical and radiological diagnostics prior to low tibial osteotomies are given. Closing wedge, opening wedge, dome-shaped, distraction, rotational and step-shaped osteotomies as well as combined procedures are described. The possibilities of these techniques are pointed out for malalignment after lower leg, pilon and ankle fractures, as well as after trauma of the distal epiphysis of the tibia. Arthroscopy of the ankle is an additional tool for detailed planning of the adequate surgical procedure. The importance of determining an early correction cannot be underestimated. PMID- 9864624 TI - [Use of growth factors in therapy of chronic wounds. Experimental, clinical and financial aspects]. AB - Clinical trials on exogenous application of polypeptide growth factors in chronic wounds have not fulfilled the high expectations derived from results of experimental studies. There is no convincing evidence that growth factors may substitute for good wound care and efficient surgical approaches to wound closure. The ultimate goal of treatment of chronic ulcerations remains reconstitution of a durable skin envelope without unstable scarring. Therefore, optimization of current methods of wound therapy, including reconstructive vascular and plastic surgery and adequate metabolic and wound control, should be employed before any adjuvant growth factor therapy is attempted. As long as efficient and inexpensive therapy of chronic wounds by growth factors has not been demonstrated, empincal growth factor treatment should be rejected on scientific and economic grounds. Current use appears to be reasonable only under a regime of controlled clinical studies comparing growth factor treatment with conventional wound therapy and operative measures according to the rules of "good clinical practice". PMID- 9864625 TI - [Textiles in the operating room. What has changed with the new medical product regulation?]. PMID- 9864626 TI - [Problems with supplementary medical care contracts]. PMID- 9864627 TI - [Hospital planning--an overview of the '90's. What are the effects on smaller hospitals?]. PMID- 9864628 TI - [Personal impressions and experiences of 2 missions in Eritrea commissioned by the Hammer Forum. Equipment and instrument supplies of operating rooms in Keren, Massawa and Asmara and subsequent possibilities for surgical interventions]. PMID- 9864629 TI - [Risk of survival of general hospitals is increasing]. PMID- 9864631 TI - [Separate care teams in the operating room]. PMID- 9864630 TI - [Tetanus/diphtheria combined vaccinations within the scope of first aid]. PMID- 9864632 TI - [Functional outcome of surgical therapy of talus fractures]. AB - Forty-five patients with fractures of the talus were treated operatively at the Department of Surgery in Bergmannsheil Hospital between 1989 and 1995. Of these, 21 patients were operated on the day of the accident. The Marti/Weber classification system was used: 9 fractures were type I, 12 type II, 18 type III and 6 type IV. Twenty-five neck-fractures were classified according to Hawkins (type I-6, type II-10, type III-6, type IV-3). The mean time of follow-up was 37.3 months (minimum of 12 months). One patient needed a primary and 2 other patients a secondary arthrodesis of the ankle and subtalar joint; 2 others required arthrodesis of the talonavicular joint. Eleven patients showed no complaints at follow-up. Twenty-five reported complaints only during strenuous exercise, 5 others during walking and 3 patients reported initial pain when walking. The mean time of therapy was 22.5 weeks. As a consequence of trauma, 7 patients changed their jobs. The most important prerequisites for successful surgery are early reduction and stable osteosynthesis. Residual complaints are frequently seen. The rate of avascular necrosis (8% in fractures of the neck of the talus) and the necessity for arthrodesis was significantly diminished. PMID- 9864633 TI - [Therapy of dislocated calcaneus joint fracture with the AO calcaneus plate]. AB - From August 1992 to March 1997, 66 patients with 71 displaced intraarticular calcaneal fractures were prospectively examined after an operative treatment using an extended lateral approach and the ASIF calcaneal plate followed by early functional postoperative treatment (mean follow-up 25 months, retrieval rate 96%). To classify the type of fracture and to verify the results of reduction and of retention CT scans in the coronal and transverse plane were performed pre- and postoperatively and on the day of assessment. The Zwipp Score was used for clinical evaluation. After fractures with 5 to 8 points according to the calcaneal fracture scale, 97% of the patients had an anatomical or near anatomical reduction of the posterior facet and the clinical outcome in 82% of the patients was graded as good or excellent. In 70% of patients with a fracture rated 9 to 10 points a good reduction was demonstrated and clinically there were 67% good or excellent results. But in the fractures with 11 to 12 points, despite 40% good reductions, the clinical outcome was graded as good in 10% of the patients only. However, if the post-operative displacement of the posterior facet was more than 2 mm no patient had a good result independent of the type of fracture. Due to restoration of the geometry of the most comminuted fracture types and the immediate partial weight bearing secondary soft tissue problems could be minimized without any loss of articular reduction. Anatomical reduction and stable internal fixation together with adequate physical therapy are apparently preconditions but not a guarantee for a good clinical result after displaced calcaneal fractures. PMID- 9864634 TI - [Baker's cyst--current surgical status. Overview and personal results]. AB - Baker's cysts were treated operatively in 19 patients in the Department of Traumatology of the University of Cologne from 1988 to 1997. The subjective and objective results were evaluated with a questionnaire and a clinical examination and sonography (follow-up: 95%). All patients were examined before surgery, sonography and X-ray of the knee were performed. We differentiated between the congenital primary cyst (39%), and the secondary form, which was always associated with an intraarticular lesion (61%). Arthroscopy was performed in all secondary forms of Baker's cyst. Postoperative complications were two reinterventions due to one hematoma and one effusion. Patient's evaluation of operation result was "excellent" in 61% and "good" in 39% of cases. All knee joints had a full range of motion. There was only one case of a recurrent cyst. The primary form of Baker's cyst has always to be extirpated, according to our clinical experience. The extirpation of the secondary Baker's cyst and the relevance of arthroscopy and treatment of the basic disease have to be discussed. PMID- 9864635 TI - [Conservative treatment after first traumatic shoulder dislocation]. AB - Between January 1989 and March 1997, 175 patients with traumatic shoulder dislocation were treated by conservative means (median age 41 years, 39 F, 136 M). In 78 patients (17 F, 61 M) a clinical and radiological follow up (median 50 months, range 6-106) could be obtained. Additionally, a diagnostic ultrasound was carried out in all patients. The recurrence rate in the group younger than 30 years (G < 30; n = 35) was 86%; in the group older than 30 years (G > 30; n = 43) it was 21% (P < 0.05). Persisting neurological deficits were found in 6 patients (8%). According to the Rowe score, 16 patients (46%) of the G < 30 achieved excellent or good results, in the G > 30, 29 patients (67%). In 17% of cases, a glenohumeral arthrosis was diagnosed be radiological means. 18% had radiological signs of a previous fracture of the greater tuberosity. Hill-Sachs lesions were identified in 19% of cases. Rotator cuff tears were diagnosed in 9% by ultrasound. No relationship between the duration of immobilization and the recurrence rate was found (P = 0.95). The recurrence rate following primary shoulder dislocation depends primarily on the patients' age. PMID- 9864636 TI - [Covering extensive soft tissue defects in infected knee endoprostheses by gastrocnemius flap]. AB - Total knee arthroplasty has become a routine procedure in surgery. Deep infections have an incidence of 2-5%. Major risk factors are large prostheses, rheumatoid arthritis, diabetes mellitus and postoperative wound-healing complications. In large soft-tissue defects with skin necrosis, local wound care shows poor results, especially if loosening of the prosthesis and necrosis of the patellar ligament are evident. In these cases, no standard surgical therapy has been developed yet. Thus, we consider meticulous debridement with synovialectomy to be mandatory. Exchange of the prosthesis may be necessary. Soft-tissue coverage ought to be performed with a gastrocnemius muscle flap covered with a split-thickness skin graft. In the last three years, 11 patients with large soft tissue defects and necrosis of the ligament were treated according to this concept. In all cases the muscle flap healed primarily and soft tissue coverage was excellent. Two patients who underwent single-stage removal and reimplantation of the prosthesis showed reinfections of the prosthesis. Reconstruction of the ligament was performed with the flap tendon. The patients with two-stage removal and reimplantation of the prosthesis and those who retained their implants had a good functional outcome. The gastrocnemius muscle flap provides easy and reliable soft-tissue reconstruction in large defects. In our patients a two-stage operation for reimplantation of the prosthesis was superior to a single-stage procedure. The reconstructed ligament should be reinforced with autologous material to prevent a secondary rupture. Early reconstruction with sufficient soft-tissue coverage and reconstruction of the ligament offers the patient the best chances of obtaining a good functional result and prevents arthrodesis or amputation. In addition, reconstructive surgery reduces the length of hospital stay and costs. PMID- 9864637 TI - [Nonbacterial osteomyelitis of the clavicle]. AB - Osteomyelitis can be subdivided into a bacterial exogenic (post traumatic/postoperative), bacterial endogenic (hematogenous) and an abacterial type, including the rare group containing primary chronic sclerosing osteomyelitis, which is typically localized in the clavicle. In a review of the literature, the criteria for the various types of chronic sclerosing osteomyelitis are analyzed. Out of a group of 17 patients with osteomyelitis of the clavicle treated between 1978 and 1996, three cases of primary chronic sclerosing osteomyelitis are demonstrated. In the differential diagnosis, primary chronic sclerosing osteomyelitis of the clavicle has to be taken into consideration despite its rareness. After establishing a diagnosis by biopsy, in contrast to the other forms, drug therapy will be the treatment of choice and not an operation. PMID- 9864638 TI - [Compartment syndrome and Volkmann contracture--can they be prevented in supracondylar humerus fracture?]. AB - The arterial blood supply of the arm is unique, as the collateral circulation is especially good. Because of this anatomic perculiarity it is possible to deal with lack of peripheral pulse without critical ischemia occurring in fractures in the elbow region. We present two children with supracondylar fractures of the humerus with damage to the brachial artery and the different management results. It is necessary to carry out the immediate diagnostic and therapeutic procedures in order to prevent late complications. We conclude that Volkmann's ischemic contracture can always be prevented. PMID- 9864639 TI - [Reconstruction of severe facial defects due to noma]. AB - Noma is an ulcerative-necrotizing gingivo-stomatitis eventually leading to severe destruction of the midface, including lips and cheek, maxilla/mandible, nose and rarely the orbit. The defects are usually unilateral. Children from economically underdeveloped countries are predominantly affected. Medically untreated the disease has a high mortality rate, which can be dramatically lowered by adequate antibiotic therapy started in time. Predisposing factors include malnourishment, immunosuppression, and poor oral hygiene. Forty-eight noma patients were surgically treated in Sokoto, Nigeria during October 1997 by an Interplast Germany team sponsored by AWD Stiftung Kinderhilfe. Interdisciplinary surgical strategies and results, e.g. ankylosis release, local flap coverage and 12 pedicled musculocutaneous latissimus dorsi island flaps, as well as a noma classification (NOMAC), are presented. PMID- 9864640 TI - [Reported malpractice after postoperative infection of the locomotor system. Analysis of 261 expert testimony evaluations]. AB - Postoperative infection in the locomotor system may lead to malpractice suits. The aim of the study was to determine the success rate of these claims and the type of expert-confirmed medical negligence so that strategies for avoiding such negligence can be offered. A total of 261 degrees concerning suspected malpractice after postoperative infection in the locomotor system were retrospectively evaluated. Of these, 43.7% claims were successful. The most frequent type of negligence was late diagnosis (34.2%), followed by inconsistent treatment (27.2%) and insufficient surgical revision (5.3%). In 33.3%, combined types of negligence were found. Most frequently, negligence was suspected after infection following hand surgery and internal fixation of long bones. The highest success rate of claims was seen after puncture of joints (69.2%). Insufficient documentation was confirmed in 15.3%. The majority of negligence resulted from diagnostic and therapeutic doubts, which could be avoided, e.g., by a clear definition of infection, consultation of a second opinion, a 2nd-look operation, undelayed and radical surgical revision, painstaking follow-up and disciplined documentation. PMID- 9864641 TI - [Removal of dirt tattooing with Q-switched laser]. AB - In present times treatment of traumatic tattoos with Q-switched laser systems is an alternative to conventional therapy. In our study, 19 patients were treated with the Q-switched ruby laser, including 4 patients with explosive trauma and 2 after pointed pencil penetration. One patient was treated with the Q-switched Nd:YAG-laser, while we tried both systems on another one. Pigmentation after abrasive trauma is easily removed or improved. Deeper penetration of foreign bodies demands more intensive and more frequent therapy. Immediate removal of foreign bodies after an accident is the best treatment. PMID- 9864642 TI - [Mutilating complex trauma of the pelvis]. AB - Traumatic hemipelvectomy is a rare, but devastating pelvic injury with few survivors reported in the literature. We report on a 19-year-old motorcyclist with a near-total hemipelvectomy. After a complicated course, the patient survived with good potential for a functional level of activity. We review the experience of other authors and give details on the management of one of the most challenging injuries confronting surgeons. Successful treatment requires extraordinary efforts and multidisciplinary team cooperation. PMID- 9864643 TI - [Interdigital pilonidal sinus ("barber's disease")--a rare occupational disease]. AB - Barber's disease is produced by short hairs that penetrate the interdigital spaces of the hands. The following chronic inflammatory reaction causes fistulas or sinuses and cysts. We present the case of a patient with interdigital pilonidal sinuses of all web spaces in both hands. Such an extensive case of this occupational disease has so far not been reported. PMID- 9864644 TI - [Nerve compression syndrome--2]. PMID- 9864645 TI - Sarcoidosis: a pediatric perspective. AB - Childhood sarcoidosis is a rare multisystemic granulomatous disease of unknown etiology. The clinical presentation can vary greatly depending upon the organs involved. Two distinct forms of sarcoidosis exist in children. Older children usually present with a multisystem disease similar to the adult manifestation, with frequent hilar lymphadenopathy and pulmonary infiltration. Early-onset childhood sarcoidosis is a unique form of the disease characterized by the triad of rash, uveitis, and arthritis in patients presenting before age 4 years. The diagnosis of sarcoidosis is confirmed by demonstrating a typical noncaseating granuloma on a biopsy specimen. The current therapy of choice for childhood sarcoidosis with multisystem involvement is corticosteroids. Methotrexate given orally in low doses is effective and safe and has steroid-sparing properties. PMID- 9864646 TI - First-morning urine specific gravity and enuresis in preschool children. AB - The object of the study was to determine whether a first-morning urine specific gravity of less than or equal to 1.015 was associated with enuresis in children 3 to 6 years old. Parents of preschool children seen at the Cleveland Clinic Foundation during a 5-month period completed a questionnaire concerning bed wetting and voiding habits of their child and collected first-morning urine specimens for specific gravity analysis. Of 101 children, 12 had a urine specific gravity of less than or equal to 1.015 (11.9%), including seven of 73 (9.6%) nonbed-wetters, two of 19 (10.5%) bed-wetters by history (who did not wet their beds on the night of the study), and three of nine (33.3%) bed-wetters (who wet their beds on the night of the study). The 73 nonbed-wetters (72.3%) had a mean (+/- standard deviation) urine specific gravity of 1.022 (+/- 0.006); 19 bed wetters by history (18.9%) had a mean urine specific gravity of 1.024 (+/- 0.006); and nine bed-wetters (8.9%) had a mean urine specific gravity of 1.019 (+/- 0.005). The groups' mean urine specific gravities were not significantly different (p = 0.10) and the enuretic children were not more likely to have first morning-void urine specific gravity of less than or equal to 1.015 than nonenuretic children (p = 0.14). Enuretic children who wet their beds on the night of the study had lower mean urine specific gravity than nonbed-wetters although the difference was not significant. Therefore, we do not recommend routine use of first warning void urine specific gravity analysis for predicting presence or absence of enuresis. PMID- 9864647 TI - Primary tracheomalacia and gastroesophageal reflux in infants with cough. AB - Cough is an uncommon sign in infants. Cough may result from the presence of abnormal secretions in the airway or abnormalities of the central airways that affect the infant's ability to clear normal secretions. Tracheomalacia (TM) and gastroesophageal reflux (GER) can both cause cough in infants. Four infants whose cough began in the newborn period were diagnosed with TM and GER. Symptoms of central airway obstruction (homophonous wheeze or tracheal cough) suggested the diagnoses. In three patients, the diagnosis was made by barium esophagraphy and airway fluoroscopy. The infants responded to conservative and medical therapy for GER and to nebulized bronchodilators. Tracheomalacia and GER cause cough in infants that begins in the newborn period. The diagnosis can often be made with studies available to the primary care provider, and the conditions are often responsive to medical management. PMID- 9864648 TI - Premature sexual development in children following the use of estrogen- or placenta-containing hair products. AB - Four African-American girls aged 14 months to 93 months developed breast or pubic hair 2 to 24 months after starting the use of estrogen or placenta-containing hair products. Discontinuing the use of the hair products resulted in regression of the breast or pubic hair. Serum gonadotropins and estradiol levels were variable. No other cause for early sexual development was noted in these girls. PMID- 9864649 TI - Infective endocarditis in the premature neonate. AB - We describe a series of 11 high-risk neonates with infective endocarditis (IE) in this retrospective review. Previously IE has rarely been diagnosed in newborns and is usually fatal. The frequency was 4.3 cases per 100 patients. Five patients survived. Microorganisms included gram positives such as S. aureus and coagulase negative Staphylococcus, gram negatives such as Klebsiella pneumoniae, Enterobacter cloacae, Enterococcus faecalis, Serratia marcescens, and Acinetobacter calcoaceticus. Echocardiographic location of the lesions showed four left sided, five right sided, and two bilateral. We conclude that IE may be more common than previously described. Prompt diagnosis and treatment led to survival in 45% of our patients. Prospective studies are needed to identify patients at risk and to establish the true incidence of IE in high-risk neonates. PMID- 9864650 TI - Neonatal endocarditis--neither rare nor fatal. PMID- 9864651 TI - Growth hormone excess in a child with neurofibromatosis type 1 and optic pathway tumor: a patient report. PMID- 9864652 TI - Tuberculosis in the home: contact history and childhood tuberculosis in central Harlem. PMID- 9864654 TI - Cost-effectiveness analysis of a rotavirus immunization program for the United States. PMID- 9864653 TI - Amoxicillin middle ear fluid penetration and pharmacokinetics in children with acute otitis media. PMID- 9864655 TI - Fulminant hepatitis associated with hepatitis A virus superinfection in patients with chronic hepatitis C. PMID- 9864656 TI - Safety and immunogenicity of heptavalent pneumococcal vaccine conjugated to CRM197 in United States infants. PMID- 9864657 TI - Pathogenesis of chronic allograft nephropathy. PMID- 9864658 TI - Haemodialysis catheter bacteraemia: evolving strategies. AB - This article reviews the new trends in the diagnosis, treatment and prevention of catheter-related bacteraemia in clinical nephrology. Among these are the newer diagnostic techniques of evaluating and obtaining culture specimens from central lines, such as timed cultures and use of the endoluminal brush. In general, attempts to salvage infected haemodialysis lines are unsuccessful. We review the data that pertain to the use of antibiotic-coated catheters in non-dialysis patients and discuss how these observations may be applied to end-stage renal disease patients. PMID- 9864659 TI - Hemofiltration and double high flux dialysis: risks and benefits. AB - Convective therapies such as hemofiltration, hemodiafiltration and double high flux dialysis have been shown to improve treatment delivered and treatment tolerance when compared to conventional dialysis therapies. The risk associated with these treatments is primarily in the quality of the substitution fluid. Technological advances now permit on-line produced substitution fluid, thereby significantly reducing the cost associated with hemofiltration and hemodiafiltration. The quality of the substitution fluid is only assured when the quality of the RO water used is within the guidelines set by the Association for the Advancement of Medical Instrumentation (AAMI). Therefore, the success of the application of this therapy is dependent on the water treatment protocols in the dialysis units. The success of this modality as a treatment for chronic renal failure is dependent on identifying those patient groups who will benefit most from this more efficient but more expensive treatment. PMID- 9864660 TI - Hemodialysis morbidity and mortality: links to patient non-compliance. AB - Patient non-compliance with the dialysis treatment prescription has often been overlooked as an important variable that contributes to outcomes in end-stage renal disease. This review discusses evidence from a limited number of researchers that such behavior may be as important as the traditional medical indicators that predict patient survival. PMID- 9864661 TI - Non-heartbeating donors: are they worth the effort? AB - This review on non-heartbeating donation focusses on three issues: the number of kidneys procured from a non-heartbeating donor programme, the transplant results and the influence of a non-heartbeating programme on public opinion regarding transplantation. PMID- 9864662 TI - New immunosuppressive strategies. AB - Discovery of novel biological and pharmaceutical agents directed against discrete molecular targets in the lympnocyte activation sequence has enabled the effective control of graft rejection by the use of combinatorial immunosuppressive therapy. Chimeric and humanized monoclonal antibodies against T-cell receptor CD3 complex chains or the IL-2 receptor block T-cell function without inducing activation, and do not cause the cytokine release syndrome of first generation products. Biological blockade of co-stimulatory molecules including CD40L and CD28 produces immunological allograft unresponsiveness in primates, though this effect is not yet proven in humans. Heterogeneity in clinical response to pharmaceutical agents is often explained by pharmacokinetic factors of absorption, metabolism and elimination. The use of microemulsion technology has increased the absorption and efficacy of cyclosporine in all organ transplants, so that there is little difference in efficacy between this agent and tacrolimus. Mycophenolate mofetil is not maximally effective alone, but significantly reduces the relative risk of acute rejection in combination with an immunophilin binding agent. It is also effective when introduced at the time of rejection. Whether it can replace other agents for maintenance immunosuppression is now under investigation. Sirolimus, the latest pharmaceutical agent to complete phase III trials, acts to inhibit IL 2 driven lymphocyte proliferation and reduces the risk of acute rejection to below 20%. Multiple pharmacokinetic interactions occur within and between these agents, so that pharmacokinetic monitoring is increasingly important. At present there are few tools to detect pharmacodynamic interactions, although reporter gene constructs and intracellular cytokine labeling offer exciting possibilities for biological monitoring. Despite these advances, none of these interventions confers demonstrable long-term benefit in graft survival or function. Acute rejection can not therefore be assumed to be a simple surrogate for chronic injury, and research must be re-focused to determine the relevant targets for long-term immunosuppression. PMID- 9864663 TI - An endothelin-transforming growth factor beta pathway in the nephrotoxicity of immunosuppressive drugs. AB - An endothelin-transforming growth factor beta type 1 pathway is proposed to account for cyclosporin nephrotoxicity. Cyclosporin amplifies the production of endothelin. Enhanced endothelin production accentuates acute vascular events and promotes the synthesis and activation of transforming growth factor beta type 1, contributing to acute and chronic pathology. This scheme integrates many observations, including the involvement of the renin-angiotensin pathway and other activators of endothelin production, and provides a rationale for the amelioration of cyclosporin nephrotoxicity. PMID- 9864664 TI - Application of newer clearance techniques for the determination of glomerular filtration rate. AB - Glomerular filtration rate (GFR) is the standard measure of renal function and is critical for the management of renal diseases. Rigorous assessment of GFR requires the measurement of renal clearance of a filtration marker, such as inulin. This method, however, is not suitable for routine clinical practice. Labelled compounds as alternative filtration markers provide accurate and precise GFR measurement, but their use may be limited for safety reasons. Thus investigators have proposed clearance procedures using minute doses of non radioactive contrast agents, including iothalamate and iohexol. Their renal clearance provides similar accuracy as inulin clearance in GFR estimation, but the need of urine collection again poses certain limitations to the procedure. Thus, plasma clearance of suitable exogenous markers, such X-ray contrast media, has been suggested for measuring renal function, in which the elimination rate of the tracer after a single intravenous injection is evaluated. Plasma clearance of these markers estimated by multiple blood samples provides precise information, but repeated sampling makes this method cumbersome. Abbreviated kinetic profiles have been proposed to predict GFR from the plasma disappearance curve. The simplified method that uses a one-compartment model corrected by the Brochner Mortensen formula gives an excellent correlation with inulin clearance and is currently employed for measuring GFR in multi-centre clinical trials. PMID- 9864665 TI - Evaluation of urinary markers in acute renal failure. AB - Acute renal failure continues to be a difficult clinical problem despite developments in dialysis and critical care. Diagnosis of the etiology frequently determines treatment. Urinalysis remains an essential diagnostic tool in the approach to acute renal failure, particularly with the current emphasis on cost containment and evidence-based medicine. This review focuses on some of the characteristic features in the urinalysis found in different forms of acute renal failure, current developments into the molecular basis for these urinary abnormalities, and new markers on the horizon. PMID- 9864666 TI - Utilization of crossmatch techniques for renal transplantation. AB - The pre-transplant cytotoxic crossmatch test is a necessary requirement for renal transplantation to proceed. However, there is no universally accepted policy on the clinical significance of a positive flow-cytometric crossmatch despite the increasing use of this technique. Differences in clinical significance found by different groups may be due to the outcome measures used and differences in the technique. The definition of a positive crossmatch can affect the results and investigators may gain information of clinical importance by considering changes in antibody levels over a series of samples from individual patients. The use of sensitive screening methods may assist in the interpretation of crossmatch results. Pre- and post-transplant crossmatching may provide data which can be used to modify immunosuppression in patients at risk of graft dysfunction. PMID- 9864667 TI - Protocol biopsies in renal transplantation: research tool or clinically useful? AB - Early protocol biopsies of stable, well functioning renal allografts reveal a high prevalence of clinically unsuspected acute and chronic pathology. It is becoming increasingly apparent that these histopathological findings are both pathogenic and predictive of long-term allograft outcome. Therefore, protocol biopsies may be required for optimal post-transplant surveillance until non invasive methods to detect allograft inflammation are developed. PMID- 9864668 TI - Endovascular stents for renal artery revascularization. AB - Revascularization cures or attenuates the clinical manifestations of renal artery stenosis (hypertension, ischemic nephropathy, pulmonary edema, angina, and congestive heart failure). Traditional approaches have been sub-optimal due to low rates of success and long-term patency after angioplasty, and to relatively high rates of perioperative morbidity and mortality. Endovascular stent placement is an alternative interventional method for renal artery revascularization. Technical success rates are excellent, and the impact on clinical outcomes (blood pressure, renal function, and cardiac complications) is promising. PMID- 9864670 TI - Dialysis and transplantation. PMID- 9864669 TI - Renal stone analysis: is there any clinical value? AB - It has usually been assumed that the analysis of recovered renal stones should be routinely done. This concept has recently been challenged. Very little work has been done to correlate analytical results with clinical findings and the impact of this information on clinical management. This paper reviews current concepts of stone formation, how this information can be used to interpret stone analysis results and how stone analyses results can be used in clinical-decision-making. PMID- 9864671 TI - Diagnostics and techniques. PMID- 9864672 TI - [Gaping eustachian tube and epitympanic retraction pockets]. PMID- 9864673 TI - [Osteoplastic frontal sinus operation with obliteration]. PMID- 9864674 TI - [Fetal surgery in the head-neck area. Current status and new possibilities with the endoscope]. AB - Fetal surgery is defined as the intrauterine surgical correction of malformations that endanger the unborn child's life in prenatal stages of development or lead to death or severe damage of the child postnatally. Such surgery is a clinical reality now. Indications for intrataurine surgical procedures also exist for head and neck abnormalities, especially in the upper respiratory tract. These include exposure and temporary obstruction of the fetal trachea for correction of pulmonary hypoplasia in cases with congenital diaphragmatic hernias, prenatal tracheotomy in cases of laryngeal atresia for the correction of lethal pulmonary overdistension, and resection of embryonic tumors that obstruct the respiratory tract. The relatively high surgical risk resulting in particular from preterm labor occurring postoperatively may be reduced by employing minimally invasive techniques. Endoscopic procedures render opening of the uterus unnecessary and are of particular importance. In part of the procedures, only endoscopic surgery has led to therapeutic success rates justifying its clinical use. Further reduction of the operative risk suggests prenatal interventions, even in cases with non-lethal conditions. More diseases of the head and neck may thus be included in the spectrum of indications. One example is prenatal correction of a cleft lip and palate, which until now has only been performed in animal experiments. The particular characteristics of fetal wound healing allow this to take place without scarring up to a certain stage in pregnancy. This offers the prospect of a surgical correction that is invisible externally and avoids growth impeding scars. The particular ethical and legal aspects of fetal surgery are discussed. PMID- 9864675 TI - [Function of the eustachian tube in epitympanic retraction pockets]. AB - There are different hypotheses for the development of epitympanic retraction pockets. These can occur as the result of an inflammatory process and negative pressure in the middle ear spaces. Anatomic bottlenecks in the middle ear favor diminished inflation of the epitympanic space. The eustachian tube provides pressure equilibration in the middle ear. Its dysfunction involving tubal opening or closing is thought to be the main reason for a permanent negative pressure in the middle ear. A patulous eustachian tube is now claimed for prolonged negative pressure in the middle ear and is caused by a so-called "sniffing" habit. The purpose of this study was to elucidate objectively the frequency of patulous eustachian tubes in ears with epitympanic retraction pockets. We examined passive tubal function in 41 patients with 50 epitympanic retraction pockets by using dual impedance in a pressure chamber. We identified three stages of epitympanic retraction pockets. Stage 1 involved a mobile and controllable retraction pocket; stage 2 corresponded with a fixed but controllable pocket, whereas stage 3 described a fixed pocket that was developing into a cholesteatoma. Results were compared with the function tests of 100 healthy ears in 50 normal individuals. We found 27 patulous eustachian tubes in patients with epitympanic retraction pockets, 22 of which occurred in affected ears while only 2 patulous tubes were found in the group of the healthy ears. Tubal opening pressure (P < 0.001) and tubal closing pressure (P < 0.01) were significantly lower in ears with epitympanic retraction pockets than in healthy ears. PMID- 9864676 TI - [Obliterating frontal sinus operation with abdominal fat. Follow-up with imaging methods]. AB - Between 1983 and 1991 ten patients with chronic frontal sinus disease underwent frontal osteoplastic flaps with fat obliteration. In order to judge operative success on a long-term basis all patients were called for postoperative follow up. They then completed a questionnaire, in which they were asked about postoperative complaints. Additionally, sinuses were examined by computed tomographic (CT) and magnetic resonance imaging (MR). Among the patients evaluated seven had no complaints, and no evidence of disease could be found by medical examination. CT scans showed complete obliteration of the frontal sinuses in all patients. In these patients, two-thirds of the fat had been replaced by connective tissue. MR was used in an attempt to show evidence for suppurative disease in the frontal sinus, but the MR images failed to correlate with symptoms. Additional results could not be obtained when compared to CT. Overall operating success could only be determined by clinical assessment and confirmed the value of the frontal osteoplastic flap with fat obliteration. PMID- 9864677 TI - [Percutaneous endoscopic gastrostomy in patients with ENT tumors]. AB - Oral food intake in patients with obstructing pharyngeal and esophageal carcinomas is commonly insufficient because of tumor-induced dysphagia which gives rise to cachexia unless treated. While entailing an unfavorable prognosis, malnutrition is often a therapy-limiting factor. Tube feeding with liquid formula diets currently offers the most efficient and least-risky approach to long-term use and is best adopted even at a pretreatment stage irrespective of the tumor therapy intended. A feeding tube placed by a percutaneous endoscopically controlled gastrostomy (PEG) increasingly offers an alternative to a nasogastric tube. After using diaphanoscopy, the stomach is punctured from outside under local anesthesia and a feeding tube inserted by means of a retrograde thread or a direct puncture method. A modification of the direct puncture method has been preferred at the Magdeburg University E.N.T. Department. The tube is held in place by thermally activated helical winding of a gastric tube end (using a memory-retaining helix). During the 1991-1996 period 415 patients with obstructing carcinomas of the upper digestive tract were treated with a feeding tube. No fatal complications were observed. Severe complications (peritonitis) occurred in three patients. In 160 patients with PEG the following parameters were recorded: weight-to-size index, body mass index, degree of dysphagia, nutrition status, lymphocyte count, total serum protein and patients' compliance to PEG. The enteral nutrition therapy used was indicated in all of the patients treated with advanced carcinomas of the head and neck. In 81% of the patients the compliance to PEG was positive. Findings demonstrated that long-term intestinal nutrition via PEG was a safe and effective form of treatment. Inserting the tube by the direct puncture method was advantageous for patients with carcinomas in the upper digestive tract as only few mechanical alterations take place along tumorous tissues following PEG while contamination with bacteria and neoplastic cells from the tumor region into the abdomen are precluded. PMID- 9864678 TI - [Neonatal hearing screening with the Echosensor automated device for otoacoustic emissions. A comparative study]. AB - Following the recommendations of the United States National Institutes of Health Consensus Conference in 1993, otoacoustic emissions (OAE) are now used internationally for hearing screening. The use of recording systems as well as the interpretation of results requires specially trained personnel although measurements are easier to perform than other recording methods available. To date, no objective method for general hearing screening, has been introduced in Germany, for neonates or children at risk, even though it would be desirable to detect and rehabilitate children with congenital hearing loss as early as possible (incidence, 1-6 per 1000). The Echosensor provides the means for carrying out OAE recordings within a short period of time and includes an automatic evaluation of results. Recordings can also be made by trained personnel. As a comparison test, audiologically trained staff recorded otoacoustic emissions in 111 new-born children in order to compare the results of the Echosensor with the results of a conventional OAE measurements device (the ILO88). The aim of this pilot study was to determine the reliability and validity of the recordings in comparison with available standards in brainstem audiometry. Our study showed that the results of the Echosensor corresponded well with the ILO88 results. Consequently, an OAE measuring method is now available is also provides high sensitivity and specificity and is easy to use. Our findings show that the Echosensor can meet the demands of systematic hearing screening in Germany. PMID- 9864679 TI - [Swallowed tools]. PMID- 9864680 TI - [Exposed superior bulb of the internal jugular vein. Differential hearing loss diagnosis with pulse synchronous tinnitus]. AB - A case report is presented of a 54-year-old woman with tinnitus synchronous with her pulse and an ipsilateral conductive hearing loss. Otoscopy showed a pulsatile structure behind the right eardrum. Imaging with high-resolution computed tomography of the temporal bone showed an enlarged jugular vein bulb. Magnetic resonance imaging could not be performed because of an implanted pacemaker. Since a glomus jugulare tumor could not be excluded right diagnostic tympanoscopy was performed and revealed a high jugular bulb that filled nearly a third of the tympanum. PMID- 9864681 TI - [Sociacusis. Non-occupationally induced hearing damage by noise, 2]. PMID- 9864682 TI - [When to prescribe hormone treatment to assure the best prevention of osteoporosis?]. PMID- 9864683 TI - [Can osteoporosis be prevented with low doses of estrogen?]. PMID- 9864684 TI - [Present status concerning hormone replacement therapy in menopause and the risk of breast cancer]. PMID- 9864685 TI - [Involvement of glucose metabolism abnormalities and insulin resistance in atherosclerotic coronary artery disease: semiquantitative coronary angiography study]. AB - Insulin resistance is known to be important in the aggravation of established coronary risk factors, but it is unclear whether insulin resistance might cause coronary artery disease as diabetes mellitus. To evaluate the relationship between insulin resistance and coronary atherosclerosis, the angiographic semiquantitative score of coronary atherosclerosis was investigated in 216 patients (175 males, 41 females) with coronary artery disease and the varying degrees of glucose metabolism including insulin resistance. Insulin resistance was estimated by plasma insulin level at 120 min after the 75g oral glucose tolerance test. Patients were divided into 3 groups according to the levels of glucose metabolism: 37 patients with treated diabetes mellitus, 48 with glucose intolerance and non-treated diabetes, and 131 with normal glucose tolerance. Multivessel disease was more frequently seen in the treated diabetes group (67.6%) than in the glucose intolerance and non-treated diabetes groups (43.8%) and in the normal glucose tolerance group (40.5%). The severity of coronary artery stenosis (8.5 +/- 3.9 vs 7.6 +/- 3.7 vs 6.6 +/- 3.7) and calcification (6.5 +/- 6.3 vs 3.3 +/- 3.3 vs 4.0 +/- 4.8) were significantly higher in the treated diabetes group than in other groups. Distal coronary lesions were significantly more frequent in the treated diabetes group (61.1% vs 17.6% vs 27.4%) than in other groups. Half of the patients in the normal glucose tolerance group had hyperinsulinemia. Multivessel disease was significantly more frequent in the insulin resistant subgroup than in the insulin non-resistant subgroup (59.4% vs 25.0%, p = 0.011). The severity of coronary artery stenosis was significantly higher in the insulin resistant subgroup than in the insulin non resistant subgroup (8.6 +/- 3.9 vs 5.6 +/- 3.0, p < 0.001), but the severity of coronary artery calcification was not significant. Distal coronary lesions showed a tendency to increase in the insulin resistant subgroup (37.5% vs 12.0%, p = 0.081). These results suggest that insulin resistance is an important risk factor for coronary artery disease in patients with normal glucose tolerance, and is related to the severity and multiplicity of coronary atherosclerosis as in patients with diabetes. PMID- 9864686 TI - [New coronary artery lesions in patients with ischemic heart disease]. AB - The appearance of a new coronary artery lesion is important in patients with acute myocardial infarction or late coronary artery bypass grafting surgery, which sometimes lead to cardiac death. We studied coronary angiograms of 1,098 patients with coronary artery disease (from 1985 to 1996) and found new lesions in 94 patients (male 74, female 20; mean age 61.6 years). Mean observation period of all 1,098 patients was 50.1 +/- 30.4 months, and that of patients with new lesions was 73.2 +/- 31.2 months. Clinical findings of the 94 patients were acute myocardial infarction (n = 28), recurrent angina (n = 56), and silent ischemia (n = 10). The appearance of a new coronary lesion was found in 2.4% (1992), 1.9% (1993), 2.3% (1994), 1.7% (1995) of all patients with ischemic heart disease per year. The new lesions were found more often in patients with non-significant stenotic lesions (n = 74) than in patients with lesions of > or = 50% stenosis (n = 20). Complex lesions were found in 56% of the recurrent angina group, and in 22% of the silent ischemia group. In our study, the rate of appearance of new coronary artery lesions was much lower than in Europe or the United States of America. These results require careful consideration for coronary revascularization procedures in Japanese patients. PMID- 9864687 TI - [Carotid artery atherosclerosis in patients with myocardial infarction]. AB - This study investigated carotid artery atherosclerosis in patients with myocardial infarction. One hundred and two patients with acute myocardial infarction [Group MI: male 86, female 16, mean 62 (range 43-79) years] and 55 normal subjects matched for age and sex with negative responses to exercise electrocardiogram testing (control group: Group C) were included. Patients in Group MI were divided into 3 subgroups according to coronary angiographic findings as follows: 57 patients with one-vessel disease (Group I: mean 59 years), 34 with 2-vessel disease (Group II: mean 64 years), and 11 with 3-vessel disease (Group III: mean 64 years). Intima-media complex thickness (IMT) of the bilateral common carotid arteries (15 mm proximal to the bifurcation) and the internal carotid arteries (15 mm distal to the bifurcation) were measured by high resolution B-mode ultrasonography, and the sum of maximum IMT of the bilateral carotid arteries (sigma IMT) were calculated. sigma IMT was significantly greater in Group MI (2.5 +/- 0.5 mm) than in Group C (1.8 +/- 0.3 mm), sigma IMT was 2.5 +/- 0.5 in Group I, 2.6 +/- 0.5 in Group II, and 2.7 +/- 0.4 mm in Group III. There was a significant positive correlation between sigma IMT and the number of involved vessels (tau = 0.45, p < 0.01). The percentage of patients with carotid artery atherosclerosis was 2% in Group C, 63% in Group MI, 54% in Group I, 68% in Group II, and 82% in Group III (p < 0.001). This study suggests that carotid artery atherosclerosis in patients with myocardial infarction is frequently complicated and severe, and more frequently complicated in patients with severe coronary artery disease. PMID- 9864688 TI - [Differences in in-hospital charge for percutaneous transluminal coronary angioplasty with and without the use of coronary stent]. AB - The cost of in-hospital percutaneous transluminal coronary angioplasty (PTCA) has risen since the introduction of the coronary stent. Increased attention is now being given to the PTCA charges in Japan and a multicenter study is necessary with regard to in-hospital charges. To clarify the differences in in-hospital charges for PTCA with and without coronary stent [Stent Group and plain old balloon angioplasty (POBA) Group, respectively], we studied the PTCA charges of 352 patients in 6 hospitals. Age, male gender and extent of coronary artery disease were not different. The ratio of acute myocardial infarction ranged from 16% to 64% and that of coronary stenting ranged from 24% to 65% (p < 0.001). In hospital charge ranged from 1.4 +/- 0.8 to 2.2 +/- 1.0 million yen (p < 0.0001). The procedural charge accounted for 53% to 75% of the in-hospital charge (p < 0.01). The in-hospital charge ranged from 1.6 +/- 0.7 to 3.3 +/- 1.6 million yen in the Stent Group, higher than the charge of 1.1 +/- 0.8 to 1.9 +/- 0.7 million yen in the POBA Group (p < 0.0001). There was a statistical difference in the number of balloon catheters used (1.1 +/- 0.4 to 2.1 +/- 0.9, p < 0.005) but not in the mean number of stents used (1.1 +/- 0.3 to 1.4 +/- 0.7). The procedural charge of the institutes with higher stenting rate (> 45%) seemed to be lower than that of the institutes with lower stenting rate (p < 0.02). In conclusion, there are large variation between institutions in PTCA charges, and in-hospital charges increased with the use of stents on introduction of the Diagnosis Related Group used in the United States. We should charge separately for coronary stenting and POBA. Despite any initial increase in the in-hospital charge for coronary stenting compared to POBA, successful stent implantation will result in a superior saving in procedural charges. PMID- 9864690 TI - [Aortic valvular vegetation in annuloaortic ectasia due to idiopathic cystic medial necrosis: a case report]. AB - Annuloaortic ectasia is often accompanied by Marfan syndrome and associated with infective endocarditis usually involving the mitral valves. We treated a patient with annuloaortic ectasia due to idiopathic cystic medial necrosis who developed congestive heart failure with aortic valvular vegetation. A 56-year-old man had dyspnea on effort since the beginning of January, 1997 and was admitted to our hospital on April 6, 1997 because of orthopnea. The diagnosis was congestive heart failure due to severe aortic regurgitation with annuloaortic ectasia detected by echocardiography. Medication and rest after hospitalization relieved his symptoms but congestive heart failure deteriorated after he had a high fever. At this time, a vegetation attached to the noncoronary cusp of the aortic valve was found which had not been detected on admission. Blood culture yielded Streptococcus sanguis. The diagnosis was infective endocarditis involving the aortic valve. Surgical correction (Bentall method) improved congestive heart failure and he was discharged on August 4, 1997 without recurrence of endocarditis. Infective endocarditis involving the aortic valves is a possible cause of development or deterioration of congestive heart failure in patients with annuloaortic ectasia. PMID- 9864689 TI - [Evaluation of therapy for dilated cardiomyopathy with heart failure by iodine 123 metaiodobenzylguanidine imaging: comparison with heart rate variability power spectral analysis]. AB - The relationship between the myocardial uptake of iodine-123 metaiodobenzylguanidine (123I-MIBG) and heart rate variability parameters has not been determined. This study determined the relationship between the change in myocardial uptake of 123I-MIBG and improvement in left ventricular function after treatment, to determine the usefulness of 123I-MIBG imaging to assess the effect of therapy on heart failure due to dilated cardiomyopathy (DCM). 123I-MIBG imaging and power spectral analysis of heart rate variability were performed before and after treatment in 17 patients with heart failure due to DCM. The following parameters were compared before and after treatment: New York Heart Association (NYHA) functional class, radiographic cardiothoracic ratio (CTR), blood pressure, echocardiographic data [left ventricular end-systolic (LVDs) and end-diastolic (LVDd) diameters, left ventricular ejection fraction (LVEF)], plasma concentrations of norepinephrine and epinephrine, heart rate variability power spectral analysis data [mean low frequency (MLF) and high frequency power (MHF)] and the myocardium to mediastinum activity ratio (MYO/M) obtained in early and late images, and washout rate calculated by anterior planar imaging of 123I MIBG. The NYHA functional class, LVEF, LVDs, CTR, MLF and MHF improved after treatment. Early MYO/M and late MYO/M improved after treatment. The rate of increase in late MYO/M was positively correlated with the rate of improvement of LVEF after treatment. Furthermore, the late MYO/M was negatively correlated with MLF. Washout rate revealed no correlation with hemodynamic parameters. These findings suggest that late MYO/M is more useful than washout rate to assess the effect of treatment on heart failure due to DCM. Furthermore, the 123I-MIBG imaging and heart rate variability parameters are useful to assess the autonomic tone in DCM with heart failure. PMID- 9864691 TI - [Suspected left ventricular outflow tract pseudoaneurysm in a patient with Takayasu's arteritis]. PMID- 9864692 TI - [Measuring the left atrial diameter with consideration given to the cardiac cycles]. PMID- 9864693 TI - [What's in the name?]. PMID- 9864694 TI - [Management of post-traumatic facial paralysis. A decision based on a series of 85 cases]. AB - OBJECTIVE: To review patient selection and factors affecting the decision for surgical or non-surgical management of patients with trauma-induced facial nerve palsy. STUDY DESIGN: Retrospective review of 85 consecutive cases occurring over a 6-year period. METHODS: Between 1984 and 1990, 85 cases of facial nerve palsy following trauma were treated. Patient assessment included clinical, audiological, radiological and EMG examinations. Depending on test results, patients were either given medical treatment or underwent total facial nerve decompression surgery. RESULTS: Recovery was achieved in all 33 medically treated patients. Among the 52 surgically-treated patients (61.2%), 47 had immediate facial nerve palsy at admission (90.4%). Onset of facial palsy was delayed in 5 others, including 3 for whom the delay to onset was unknown. Surgical accesses used were the middle fossa and transmastoidal (60%), translabyrinthine (11%), and pure transmastoidal (29%) routes depending on hearing loss, location of fracture line and general patient condition. Lesions were predominantly found in the geniculate ganglion area. A nerve gap was found in only 13.5% of the cases. Two years post-operatively, 92.3% of the patients had grade I-III recovery. No grade V or VI cases were observed. DISCUSSION AND CONCLUSION: The rarity of severe nerve lesions encountered in surgically-treated patients raises the question of better candidate selection for surgery. At present, imaging techniques, particularly radiology, can help to choose the best indication. Immediate total facial palsy associated with a clear-cut fracture going through the Fallopian canal is perhaps the only case requiring surgery. When the delay of onset is unknown, an EMG indicating a total denervation process is the decisive argument. PMID- 9864695 TI - [Chronic bilateral subdural hematomas]. AB - BACKGROUND AND PURPOSE: Are chronic bilateral subdural hematomas different from unilateral forms in terms of delay of diagnosis, clinical presentation and post operative recovery or recurrence? METHODS: Etiological, clinical, and radiological aspects and management outcome of bilateral chronic subdural hematomas consecutively managed from 1990 to 1995 were retrospectively analyzed and compared with unilateral forms managed in the same neurosurgical unit during the same period of time. RESULTS: There were 236 cases. Bilateral subdural hematomas occurred in 41 patients (17.4%). A good outcome was obtained in 97.5% of the cases and morbidity was 14.6%. The differences significantly observed between unilateral and bilateral subdural hematomas were duration of the prediagnostic period, pneumatoceles and recurrence rates (12%). Recurrences were all observed during the first postoperative month and treated with the same initial standardized procedure. The rate of infection or epileptic seizures was low. CONCLUSIONS: There are few limited differences between uni- and bilateral chronic subdural hematomas. In the latter, a possible correlation between the duration of prediagnostic period and an increased rate of recurrence could be explained by poorer cerebral reexpansion. The overall prognosis of bilateral hematomas is comparable with that observed in unilateral forms. A simple and routinely used burr hole procedure either under local or general anesthesia associated with closed drainage is a safe surgical procedure for either bilateral or unilateral forms and can be performed in all the cases including recurrences. Systematic antiepileptic drugs or antibiotherapy are unnecessary in the medical management of chronic subdural hematomas. One late delayed postoperative clinical control seems to be sufficient. PMID- 9864696 TI - [Oculomotor paralysis and head trauma]. AB - Few studies have dealt with post-trauma ocular motor palsy. We analyzed 31 personal cases and reviewed the literature, focusing on pathogenesis. We emphasize the importance of a complete radiological examination including magnetic resonance imaging when the CT-scan does not provide a satisfactory correlation between anatomic findings and clinical presentation. Systematic detection, early management and regular follow-up is mandatory. PMID- 9864697 TI - [Von Hippel-Lindau disease and central nervous system hemangioblastoma. Progress in genetics and clinical management]. AB - Von Hippel-Lindau (VHL) disease is an autosomal dominant disorder, predisposing to the development of various tumors (central nervous system and retinal hemangioblastomas, endolymphatic sac tumors, renal cell carcinoma and/or renal cysts, pheochromocytomas, pancreatic cysts and/or tumors). Incidence of the disease is 1/36,000. Central nervous system hemangioblastomas and renal cell carcinoma are the main causes of death. The VHL gene, located on chromosome 3p25 26, is a tumor-suppressor gene encoding for a 213 amino acid protein which plays a major role in regulation of VEGF expression. Germline mutations of the VHL gene are identified in about 75-80% of the patients. Somatic mutations of the VHL gene are found in both sporadic central nervous system hemangioblastomas and sporadic "clear" renal cell carcinomas. For neurosurgeons search for VHL disease should be imperative in presence of a patient with apparently "sporadic" central nervous system or endolymphatic sac tumor. PMID- 9864698 TI - [Neurofibromatosis]. AB - Neurofibromatoses regroup at least two different autosomal dominant disorders, neurofibromatosis type 1 (NF1) and neurofibromatosis type 2 (NF2). NF1 gene and NF2 genes have been respectively localized on chromosomes 17 and 22. NF1 represents 95% of neurofibromatoses cases. Its incidence is 1 for 3,500 newborns, its prevalence 1 for 4,500. NF1 is characterized by its cutaneous manifestations, cafe au lait spots, lentigines and neurofibromas. NF2 incidence is 1 for 33,000 40,000 newborns. NF2 is characterized by bilateral vestibular schwannomas (former acoustic neurinomas) and other tumors of the central nervous system. The hallmark of schwannomatosis (neurilemmomatosis) is multiple cutaneous or subcutaneous schwannomas without vestibular schwannomas. Different neurofibromatoses are characterized by different prognosis, complications, and genetic counseling. Multidisciplinary centers with trained physicians are ideal structures for management of neurofibromatoses patients. PMID- 9864699 TI - [Guidelines for the maintenance of Hippel-Lindau disease in daily practice. French Society of Neurosurgery]. AB - The subject of the Teaching Course organized by the French Society of Neurosurgery during its Winter meeting in 1998 was phacomatosis. Special interest was given to the Von Hippel-Lindau (VHL) disease. Guidelines for the management of this disease are the following: The diagnosis is established in the presence of two major lesions, or with one major lesion associated with a familial history, or when a VHL gene mutation is detected. An isolated hemangioblastoma (HBM) is associated with the disease in 36% of the cases when located in the posterior fossa and in 64% of the cases when located in the spinal cord. The main causes of mortality of VHL are the following: HBM (61%), renal cancer (31%) and pheochromocytoma (PCC) (8%). The percentage of PCC in the French series is 24%. The percentage of Endolymphatic tumor in the French series is 4%. No surgery is recommended in case of asymptomatic HBM (but radiosurgery has to be evaluated). Follow-up requires regular clinical examinations, one ophthalmological examination per year, one abdominal ultrasonography per year, and one cerebral MRI every three years. PMID- 9864700 TI - [Long-term survival after metastatic cerebral melanoma]. AB - BACKGROUND AND PURPOSE: Long-term survival, i.e. more than 10 years after the surgical removal of a metastatic intracerebral melanoma, is very rare. We report such a case, the patient is living and in good health 12 years later. CASE REPORT: In 1985, a 63-year-old woman underwent occipital craniotomy. An ovoid, 4 cm diameter, superficial, black tumor was removed. The diagnosis of a metastatic cerebral melanoma was confirmed by histopathology and immunocytochemistry. Ten years before she had had a superficial spreading melanoma excised from her right foot. She has recently been seen without any recurrence of her melanoma. CONCLUSION: Three other cases of long-term survival after metastatic cerebral melanoma have been reported. Additionally, our patient had her metastatic cerebral melanoma ten years after removal of a superficial spreading melanoma. There are only a few examples of such long intervals before the occurrence of metastatic cerebral melanoma. Sometimes, the past medical history of a patient with cerebral metastasis should be investigated very carefully. PMID- 9864701 TI - [Intra-orbital dacryops. Report of a case]. AB - We report a case of intra-orbital Dacryops in a 7-year-old child. The presenting sign was progressive exophtalmus. The patient was operated via a lateral approach allowing total removal of the cyst without recurrence after one year. The clinical, radiological (CT Scan and MRI) and therapeutical aspects are discussed. PMID- 9864702 TI - [Epidermoid cyst of the third ventricle. Report of a case]. AB - We report a case of epidermal (or epidermoid) cyst, in a 36-year-old man which developed in the third ventricle. Clinical manifestations were headaches and memory disturbances. On CT scan the tumor occupied the entire third ventricle but was mainly developed on the left side. On CT reconstructed images, the floor of the third ventricle was clearly visible. Using a trans-ventricular approach, the tumor, closely related to the left part of hypothalamus, was totally removed. Later on, because of persistent hydrocephalus, a ventricular shunt was inserted. An aseptic meningitis occurred and resolved spontaneously. The patient exhibited a postoperative transitory Korsakoff's syndrome. Postoperative endocrine investigations showed hypopituitarism. Some intra-ventricular epidermal cysts have been reported, especially involving the fourth ventricle. Their development into the third ventricle is unusual, and in early reports their precise origin appears doubtful. Although they have no characteristic radiological features, the location of epidermal cysts is clearly defined by the CT scan and especially MRI. It would be possible to totally remove epidermal cysts of the third ventricle, avoiding the risk of recurrence. PMID- 9864703 TI - [Approach to severe functional deficiencies: evaluating the efficacy of a special multidisciplinary consultation]. AB - A global approach is required to evaluate severe functional deficits. We have developed a multidisciplinary consultation and assessed its usefulness after one year. A total of 62 patients were evaluated (mean age 35 years), usually for neurological functional deficit (50%). Among the 62 patients, surgery was proposed in 72.6% (operations performed in 66%). Others were managed with non surgical procedures or abstention. The functional outcome as assessed by patients and physicians was poor (or no result) for 6%, fair for 15%, good or very good for 79%. These results suggest that such a multidisciplinary approach is worth being pursued. Some adjustments for psychological dysfunction may further improve outcomes. PMID- 9864704 TI - [Application of guidelines concerning the decontamination and sterilization of instruments used in the neurosurgery operating theater and the risk of contamination from Creutzfeldt-Jakob disease. French Society of Neurosurgery]. PMID- 9864705 TI - [Spatial organization of the cauda equina]. PMID- 9864706 TI - [Daily excretion of epinephrine and norepinephrine in acute phase of cerebral ischemia]. AB - The aim of the study was to assess the relation between urinary excretion of epinephrine and norepinephrine, and stroke severity, prognosis and standard biochemical investigations in acute phase of ischaemic stroke. The study was done on the material of 36 patients (17 women and 19 men, aged 41-89 years, mean: 69 +/- 13 years) admitted to the stroke unit of the Neurology Department within 24 hours (mean: 7.7 hours), after the first ever ischaemic stroke affecting the territory of the internal carotid artery circulation, confirmed by CT. Patients with the history of diabetes mellitus, cardiac insufficiency and infection on admission were excluded from the study. The 24-hour urine collection for catecholamines (epinephrine and norepinephrine) was done on the first and third day of hospitalization. An even single increased daily excretion of epinephrine (> 10 micrograms/24 hr) was found in 21 and norepinephrine (> 50 micrograms/24 hr) in 10 of 36 patients. No correlation between daily excretion of epinephrine and norepinephrine was found. Statistical analysis revealed a significant relation between norepinephrine, but not epinephrine, and neurological deficit, prognosis and some biochemical parameters of investigated patients. Patients with increased daily excretion of norepinephrine showed worse neurological deficit (p < 0.01), greater mortality assessed on the 30-th and 90-th day of stroke (p < 0.05) and increased plasma glucose, sedimentation rate, white blood cells count, creatinine kinase activity as well as microalbuminuria and decreased plasma kalium concentration. PMID- 9864707 TI - [Acute hypercapnia increases the permeability of blood/cerebrospinal fluid barrier for gentamicin in rabbits under general anesthesia]. AB - The aim of the work was to demonstrate whether acute hypercapnia (paCO2 > 65 mmHg) influenced the permeability of BBCSF. 12 Chinchilla rabbits which underwent general anaesthesia were randomly divided into 2 groups. To maintain normal paCO2 values, the investigation was performed under normal ventilation in control group (No I). Controlled hypoventilation was applied to achieve acute hypercapnia in the shortest possible time in the investigated group (No II). BBCSF function was evaluated using gentamicin permeability indexes QG (defined as gentamicin concentration in the cerebrospinal fluid-to-serum gentamicin concentration in given moment of investigation). Mean haemodynamic and haematologic parameter values showed no significant within-group and between-group deviation. Statistically insignificant increase of QG values after the first hour of hypercapnia was found when comparing mean QG values in the investigated group. Mean QG value was statistically significantly higher in the animals hypoventilated for 3 hours when compared with animals which underwent normal ventilation during the whole procedure. Severe, acute hypercapnia disturbs the integrality of the blood/cerebrospinal fluid barrier. PMID- 9864708 TI - ["Repty" stroke scale . "Repty" Scale for the evaluation of the degree of injury after cerebral stroke. Part I]. AB - Own modification of stroke scale, which enables evaluation of impairment in hemiplegic patients is presented. The scale named "Repty" consists of 15 items, including bowel and bladder function, enables precise evaluation of neurological impairments. The comparison between "Repty" scale and Orgogozo scale was carried out. Interobserver agreement and correlation between both scales was calculated. PMID- 9864709 TI - ["Repty" scales of strokes. Functional index "Repty" for the evaluation of ADL in hemiplegic patients after cerebral stroke. Part II]. AB - Own modification of the Functional Independence Measure, which enables evaluation of activities of daily living in hemiplegic stroke patients is presented. The index of function "Repty", shorter and simpler, consists of 15 items, and makes possible exact estimation of functional state. This is useful in evaluation of the effects of treatment, and rehabilitation, and state of disability as well. The advantage of the "Repty" index over Barthel index results from evaluation of verbal communication and more extended scoring system. PMID- 9864710 TI - [The influence of diabetes on neurological outcome and mortality in stroke patients]. AB - In a retrospective study of diabetic and non-diabetic patients with acute ischaemic stroke, mortality and neurological outcome were assessed. Mortality was determined 4 weeks after admission. The study population consisted of the consecutive patients (n = 609) admitted to the Department of Neurology with acute stroke between January 1994 and December 1995. There were 80 diabetic and 529 non diabetic patients. The mortality rate was higher in diabetics (36.3%) than in non diabetics (27%), but the difference was not significant (p = 0.08). The neurological status on admission, age, history of arterial hypertension, atrial fibrillation and coronary arterial disease were used to select groups of 41 diabetics and 84 non-diabetics with similar risk factors and similar onset of stroke. The neurological status on 10-th, 20-th and 28-th day of the hospital stay in both groups was compared. The study showed that insulin-dependent diabetics and older patients (above 65 years of age) with diabetes demonstrated a worse neurological outcome than non-diabetic patients, although the difference was not significant. PMID- 9864711 TI - [Assessment of the efficacy of treatment with pimozide in patients with amyotrophic lateral sclerosis. Introductory notes]. AB - The aim of the study was to assess the effect of pimozide voltage-dependent calcium channel blocker on the progression of ALS patients as compared to the potentially neuroprotective drugs, selegiline and vitamin E. There were 44 patients (17 females and 27 males, aged from 30 to 80 years, mean age: 56.2 years) diagnosed as either definite or possible ALS. The study design was open randomised. Patients were treated 3-12 months; the daily dose of pimozide was 1 mg. The disease progression index was calculated as a difference between scores of Norris scale before and after treatment. Statistical analysis showed a significant decrease of the index of progression of the disease in pimozide treated patients as compared to the others. This effect was neither related to the progression of the disease nor advance of the disease at the beginning of treatment. PMID- 9864712 TI - [Metastatic vertebral tumors diagnosed by percutaneous needle biopsy]. AB - The purpose of this paper is to establish diagnostic value of percutaneous needle biopsy in patients with vertebral tumours treated in Department of Neurosurgery in Poznan between 1975 and 1996. Radiological examinations: X-rays, plain X-rays, CT and MR, demonstrated vertebral destruction and allowed needle biopsy performance. The most frequent destruction was observed in thoracic region (45% of cases), rarely--in sacral bone (5%). In cases with single neoplastic focus, only in vertebral body, histological diagnosis by needle biopsy was achieved in 70% of cases. However in patients with paravertebral tumour positive results of needle biopsy were higher--82% of cases. PMID- 9864713 TI - [Transpedicular stabilization in spinal and vertebral column injuries in thoracic and lumbar spine segments]. AB - The authors present their own experience in application of transpedicular internal stabilization of the thoracic and lumbar spine. Clinical analysis was carried out in a group of 12 patients after vertebral column and spinal trauma managed surgically in the Department of Neurosurgery in Poznan between 1.06.95 and 31.12.96. Age of patients ranged from 19 to 56 years (mean age 35.08 +/- 13.04 yrs.). The level of vertebral fracture was as follows: thoracic (2 cases), thoraco-lumbar (6 cases) and lumbar (4 cases). Three patients were completely paraplegic. All patients underwent posterior or posterolateral surgical approach. The fractured parts of bones, translocated into vertebral canal were removed and nervous structures were decompressed. Transpendicular stabilization was performed after the decompression. Improvement of neurological condition was observed in 8 patients. PMID- 9864714 TI - [Reconstruction methods in skull base surgery]. AB - Surgery for extensive skull base lesions inevitably creates multiple tissue defects that require reconstruction. In this paper reconstruction methods applied in 41 skull base surgery procedures are presented. Anterior cranial fossa was involved in 13 operations (4 malignant and 9 non-malignant tumours), middle and posterior cranial fossa in 28 (9 malignant and 19 non-malignant). As a rule all the defects were reconstructed in one stage, immediately after tumour removal. Titanium mesh was the only alloplastic material used to cover bone defects in 7 patients. Otherwise only autografts were used. Postoperatively in all the patients in whom excision and repair of dura was necessary, continuous lumbar drainage of cerebrospinal fluid was maintained for 4-6 days. In anterior cranial fossa area large pediculated, vascularized periosteal flap allowed for reliable separation of upper airways from intracranial cavity. Titanium mesh was used to cover orbital wall defects. In the area of middle cranial fossa and temporal bone the most important aspects of reconstruction were watertight closure of the dura, separation of the surgical cavity from nasopharynx with rotated temporalis muscle, and facial nerve reconstruction (the nerve was reconstructed in 7 cases). Additionally in 1 patient hypoglossal nerve was sutured end-to-end. In 2 patients the invaded segment of internal carotid artery was excised and replaced with prosthesis. In 13 patients the surgical procedure was performed in an area that was previously operated upon and/or irradiated. Delayed healing was seen in 6 cases, but none required additional procedures and the wounds were completely healed with local treatment. Reconstruction methods presented in this paper are relatively simple and do not necessitate modern materials. Even in extensive procedures proper wound healing and satisfactory cosmetic result were obtained. PMID- 9864715 TI - [Occipital nerve neuralgia as postoperative complication. Views on etiology and treatment]. AB - Occipital neuralgia might have a various etiology but the most common cause is hypertrophic fibrosis of subcutaneous tissue following trauma to the occipital region, surrounding the occipital nerve. The other important cause of neuralgia is spondylosis of the upper part of the cervical spine (C1-C2). Rare causes are diabetes, gout and neoplasm. In the presented material we point out the possibility of the occipital neuralgia after surgery in the treatment of the tumours of ponto-cerebellar region and trigeminal neuralgia. We observed the symptoms in two groups of patients and used pharmacological treatment, local blockade and cutting (neuronectomy) of the occipital nerves trunk when the conservative treatment was unsuccessful. We also present the present concepts of occipital neuralgia treatment. In case of severe symptoms, unsuccessful conservative therapy and poor results of the neuronectomy the most indicated therapy is selective posterior rhizotomy. PMID- 9864716 TI - [Expression of p53 protein and proliferative activity in multiform glioblastoma]. AB - The aim of the study of 41 multiform glioblastomas was the analysis of p53 protein immunoreactivity in neoplastic cells and evaluation of relationship of this biologic marker to tumour proliferation activity. Positive p53 expression was observed in 24 (58.5%) tumours, the negative one in 17 tumours (41.5%). Proliferation indexes of PCNA, anti-Ki6 and AgNORs showed high values in the multiform glioblastoma p53 positive group, but without statistical differences in comparison with the group of p53-negative glioblastomas. Significant differences were observed in survival time of patients with p53 positive tumours in comparison with p53-negative ones. In 15 patients with p53-positive multiform glioblastomas survival time was less than 6 months (62.5%) on the contrary with only 4 patients with similar survival time in p53-negative glioblastoma group (23.5%). Our results suggest that p53 expression in multiform glioblastoma cells, generally considered as the indirect index of p53 suppressor gene, reflects aggressive stadium of neoplastic disease and significantly worsens the prognosis. PMID- 9864717 TI - [Role thrombocytes in etiopathogenesis of ischemic stroke]. AB - This paper presents the most essential problems concerning the role of platelets in the genesis of ischaemic brain disease and their role in spreading of ischaemic focus. The role of platelets in atherogenesis and development of vascular abnormalities in arterial hypertension and diabetes mellitus was considered, and attention was paid to conditions concerned with the pathogenesis of ischaemic stroke such as presence of antiphospholipid antibodies and presence of large volume platelets, in which platelet activation plays essential role. Then it was shown that platelet hyperactivation occurs in acute phase of ischaemic stroke and mechanisms of destructive action of activated platelets on recent ischaemic focus in brain such as secretion of neurotoxins and blocking of microcirculation by platelet aggregates were discussed. PMID- 9864718 TI - [Current problems in myotonic dystrophy]. AB - Myotonic dystrophy (m.d.) is an autosomal dominant multisystem disorder involving muscles, brain, heart, eye, endocrine system, alimentary and respiratory systems. M.d. is the most frequent cause of muscle dystrophy. Unstable CTG trinucleotide repeat at 3' untranslated end of the myotonic protein kinase gene on chromosome 19q 13.3 is the molecular basis of the disease. Normal length of CTG trinucleotide repeat is 5-40. Molecular mechanism of the myotonic dystrophy is discussed. Cataract, heart dysfunction, endocrine organs dysfunction, gallbladder stones, impotence are characteristic changes in patient with m.d. Apathy, drowsiness, sometimes dementia point to central nervous system involvement. Clinical course, correlation between CTG expansion and clinical manifestation are described. Nowadays progress in molecular genetic allows to make the diagnosis by DNA examination. Prenatal diagnosis is also possible. PMID- 9864719 TI - [Spontaneous dissection of cerebral arteries. I. Carotid arteries and their branches]. AB - Artery dissection means tearing apart of its layers by blood coming inside after endothelial damage. The authors describe the prevalence of that pathology of carotid, arteries (diagnosed rarely so far), pointing to early onset age and to its multifactorial aetiology and pathogenesis. They also outline the pathomechanism of the neurological symptoms and emphasize the variety of the clinical manifestations. The diagnostic possibilities of artery dissection are also presented. PMID- 9864720 TI - [Huntington chorea in a boy aged 14 years]. AB - The authors present a case of Huntington disease in a 14 year old boy. The basis of genetic, pathoetiology, clinical course of disease, significance of molecular analysis of DNA are discussed. PMID- 9864721 TI - [Spontaneous brain stem hemorrhage: analysis of cases]. AB - Four cases of spontaneous haemorrhages to the brain stem were described. They concerned young patients (middle age was 34 years) without previous risk factors. The diagnosis was established on the ground of clinical examination, neuroimaging methods-CT, MR, cerebral angiography. We could not indicate the cause of haemorrhage (i.e. a blood vessel pathology, nor coagulation disturbances neither other pathological coincidence were found). All patients survived, but none of them is able to continue his professional work. Two patients demonstrate slight pyramidal signs, and slight hemihypaesthesia. One patient still presents profound brain stem symptoms, and in the last case tetraparesis remains. PMID- 9864722 TI - [Juvenile monomelic amyotrophy: Hirayama disease]. AB - We present three patients with unilateral upper limb weakness (with muscular atrophy)-two of them with distal and one with proximal localization. The disease onset was between 18th end 35-th year of life; the disease course was biphasic (i.e. progressive within first 1 to 3 years, and stabilized during following 4-24 years). The laboratory investigations permitted to diagnose juvenile monomelic amyotrophy, an entity that is very rare outside Japan. Electromyography revealed neurogenic involvement with spinal features also in clinically unaffected muscles. We suggest that these results may support the hypothesis of this disease being a benign variant of spinal muscular atrophy. PMID- 9864723 TI - [Hemorrhagic manifestation of moyamoya disease in a 19-year-old woman with mental retardation]. AB - The paper presents a case of moyamoya disease, with first clinical sign subarachnoid haemorrhage in a 19-year old woman with mental impairment. The CT screen shows the presence of ischaemic areas of haemorrhagic type. MR, angio-MR and classical angiography confirmed the presence of ischaemic areas with contraction of the distal parts of the internal carotid arteries and left anterior cerebral artery. Authors call attention to the diagnostic problems of the case and accent the very important role of MR and MR angiography in the diagnosis of moyamoya disease. PMID- 9864724 TI - [A trial of progressive supranuclear palsy with carbamazepine]. AB - We present a 61-year old patient with diagnosis of progressive supranuclear palsy. 2 years after the beginning of her complaints the diagnosis was based on the results of neurological examination. The ENG examination (among others vertical gaze paralysis) and the MR examination (2 of 3 criteria were met). The course of the disease was complicated by epilepsy and the treatment with carbamazepine was commenced. A significant improvement of her neurological and mental status was observed. The authors suggest checking this observation on larger number of patients with PSP. PMID- 9864725 TI - [Own experience in surgical treatment of intramedullary spinal cord lipomas]. AB - The aim of this report is to present own experiences in surgical treatment of extremely rare occurring intramedullary spinal cord lipomas without spinal dysraphism. Our experience with such tumours based on 2 cases out of 15 intramedullary spinal cord tumours that were treated at our department from 1984 to 1997. The tumours were located at C4-C7 and C6-Th6 spinal cord region. The patients were 16-year-old woman and 57-year-old woman. Both patients presented with long history of progressive spinal cord damage with rapid deterioration after mild injury. The diagnosis was based on myelography in one case and on MRI in the second one. In first case the tumour was totally removed and we achieved neurological improvement. In the second case the tumour was subtotally removed and after 12 months on MR examination we found relapse of the tumour without neurological deterioration. We found two types of the tumours. The first one was well separated from the normal spinal cord and easy to remove. In the second it closely adhered to the spinal cord without possibility of total excision. The surgical treatment of intramedullary spinal cord lipomas is very difficult and dangerous but only in this way we can protect the patients from total paralysis and to improve their comfort of the life. PMID- 9864727 TI - [ A case of parasellar tumor with double histological texture: chromophobe pituitary adenoma and epidermoid cyst]. AB - A rare case of a 42-year-old woman with an intra- and suprasellar tumour with double histological texture (chromophobe pituitary adenoma and epidermoid cyst) is presented. Elevated serum prolactin levels were found on admission. After the surgery symptoms of diabetes insipidus appeared. On the ground of the presented case and review of references it seems that the coexistence of an epidermoid cyst and a pituitary adenoma suggest their common histogenetic origin. PMID- 9864726 TI - [Acute hematoma between the dura and the external membrane of hematoma capsule of after the removal of bilateral chronic subdural hematomas by trepanation. Case report]. AB - A case of reported of complication following surgical treatment of bilateral subdural haematomas in an elderly man. Acute haematoma developed after the operation between the dura and the external surface of the capsule of the haematoma which had been removed several hours earlier through a trepanation hole. Attention is called to the atypical localization of the complication with bleeding externally to the capsule and not into the space left after haematoma evacuation which is usually observed in case of haematoma recurrence. The problem is discussed, whether such bleeding during chronic persistence of haematoma can lead to the development of multilayer and multilocular haematomas. In the presented case quick recognition of the complication followed by reoperation with removal of haematoma and bleeding capsule through craniotomy with rehabilitation treatment after the operation has led to very good therapeutic effect. PMID- 9864728 TI - [Subdural hematoma with recurrent syndromes simulating TIA]. PMID- 9864729 TI - Enhancement of spatial-temporal reasoning after a Mozart listening condition in Alzheimer's disease: a case study. AB - Several recent studies have investigated the effectiveness of various behavioral interventions on the cognitive performance of subjects with Alzheimer's disease (AD). Simulations of Shaw's structured model of the cortex led to the predictions that music might enhance spatial-temporal reasoning. A subsequent behavioral study in college students documented an improvement in scores on a spatial temporal task after listening to a Mozart piano sonata. In this study, we investigated the enhancement of scores on a spatial-temporal task after a Mozart listening condition in a set of twins who are discordant for AD. After listening to an excerpt from a Mozart piano sonata, the AD twin showed considerable improvement on the spatial-temporal task when compared with pretest scores. Furthermore, no enhancement of scores was seen following either of the control conditions (i.e., silence or 1930s popular tunes). This finding suggests that music may be used as a tool to investigate functional plasticity in Alzheimer's disease and to better understand the underlying pathophysiology. PMID- 9864730 TI - Comprehensive management of arteriovenous malformations. AB - The treatment of arteriovenous malformations depends on the efforts of a multidisciplinary team whose ultimate goal is to achieve better results when compared to the natural history of the pathology. The role of adjuvant treatment modalities such as radiosurgery and endovascular embolization is discussed. Treatment strategies and surgical results from a personal series of 344 patients operated in a ten-year period are reviewed. The Spetzler and Martin classification was modified to include subgroups IIIA (large size grade III AVMs) and IIIB (small grade III AVMs in eloquent areas) to assist the surgical resection criteria. The treatment strategy followed was surgery for grades I and II, embolization plus surgery for grades IIIA, radiosurgery for grades IIIB, and conservative for grades IV and V. According to the new proposed classification 45 (13%) patients were grade I, 96 (28%) were grade II, 44 (13%) grade IIIA, 97 (28%) grade IIIB, 45 (13%) grade IV, and 17 (5%) were grade V. As for surgical results 85.8% of the patients had a good outcome (no additional neurological deficit), 12.5% had a fair outcome (minor neurological deficit), 0.6% had a bad outcome (major neurological deficit), and 1.2% died. These figures indicate that the treatment of arteriovenous malformations can achieve better results compared to the natural history if managed by a well trained group of specialists led by an experienced neurosurgeon. PMID- 9864732 TI - Temporal profile of adenovirus-mediated E. coli lacZ gene expression in normal and post-ischemic gerbil hippocampus and ventricle. AB - A replication-defective adenoviral vector containing the E. coli lacZ gene was directly injected into normal and post-ischemic gerbil right hippocampus and lateral ventricle, and temporal profiles of the exogenous gene expression were compared. In case of ischemia, common carotid arteries (CCA) were transiently occluded for 5 min, and the adenoviral vector was administered just after the reperfusion. The animals were recovered for 8 h, 1, 3, 7 or 21 days. A small to moderate number of neural cells in the normal hippocampus expressed the gene from 1-3 days except for the cells around dentate gyrus (DG) and the needle route that began to express from 8 h of injection. Some normal hippocampal cells persisted the expression until 7 days. A moderate to large number of ventricular cells expressed the lacZ gene from 8 h to 7 days in the normal brain. On the other hand, no expression of the lacZ gene was observed in the post-ischemic hippocampus at 8 h including cells at DG and the needle route. Hippocampal CA1 neurons, that were selectively lost at 7 days of reperfusion, never expressed the gene throughout the post-ischemic course. The other hippocampal cells such as CA3 and dentate granule cells that survived ischemia expressed the gene only transiently at 1 day. A robust expression of the gene persisted in the ventricular cells from 8 h to 7 days. The majority of brain cells in the hippocampus that expressed the lacZ gene was not the pyramidal neurons, but small neurons at around the pyramidal layers of DG. Some astroglial, but no microglial, cells expressed the lacZ gene in the hippocampus. The present study shows that an expression of the lacZ gene was limited in the post-ischemic gerbil hippocampus especially at the vulnerable CA1 layer in contrast to the strong and persistent expression in the ventricular cells, and that the majority of beta-gal positive cells were not the pyramidal neurons but small neurons at around the cell layer both in the control and post-ischemic gerbil hippocampus. PMID- 9864731 TI - Japanese Creutzfeldt-Jakob disease patients exhibiting high incidence of the E200K PRNP mutation and located in the basin of a river. AB - Seven cases with Creutzfeldt-Jakob disease (CJD) located in the basin of the Fuji river (Fuji area) in Japan were examined genetically and clinicopathologically. The onset of the disease was between 1989 and 1995. All cases were from different families, although 3 cases were family members of previously reported CJD patients. They had clinical and/or neuropathological features, corresponding to subacute spongiform encephalopathy. Five of the 7 cases, including the 3 familial cases, had the E200K mutation in the gene encoding prion protein (PRNP). It is suggested that there is a small cluster of CJD patients with a founder effect of the E200K mutation in the Fuji area, because the incidence of CJD with the E200K mutation appears to be much higher in this area than other areas in Japan. The disease penetrance of the 5 cases with the E200K mutation seems to be low, and they may have an age-related incidence in the Fuji area. These findings support the hypothesis that the phenotypes of CJD patients with the PRNP mutations are linked to the position of the mutation, but not related to ethnic or environmental factors. PMID- 9864733 TI - The microglial response to progressive hydrocephalus in a model of inherited aqueductal stenosis. AB - Although gliosis has been reported to be a common and persistent feature in the white matter of hydrocephalic brains, no studies have identified the cell types that characterize this response. Therefore, the present study has employed histochemical methods to evaluate microglial cells in the brains of infant rats with inherited hydrocephalus. This strain of rats acquires hydrocephalus during late fetal stages due to aqueductal stenosis. Tissue from the sensorimotor and auditory cortices of 12- and 21-day-old hydrocephalic and normal H-Tx rats was processed and stained for the lectin microglial marker Griffonia simplicifolia (GSA-IB4). During the progression of hydrocephalus, GSA-positive cells exhibited three changes: (1) Cytologically, the cell bodies were enlarged, and their processes were thicker, longer and more numerous. These changes were most notable in the gray matter. (2) The packing density of GSA-positive cells was either increased or decreased, depending on the age of the animal and the severity of hydrocephalus. (3) Localized clusters of GSA-positive cells were conspicuous in the white matter of 12-day animals with mild hydrocephalus, and in the gray matter of 21-day animals with severe hydrocephalus. These results indicate that the microglial response is initiated during intermediate stages of hydrocephalus, and is not restricted to the periventricular white matter. These changes may signal other pathophysiologic events in the hydrocephalic brain, and demonstrate that microglia constitute one important element in the gliosis that accompanies hydrocephalus. PMID- 9864734 TI - The utility of contrast-enhanced MR-angiography for posterior fossa giant cerebral aneurysm management. AB - The utility of magnetic resonance angiography is sometimes limited in the diagnostic workup of cerebral aneurysms with low flow and/or partial thrombosis when weighed against digital subtraction angiography. We present the case of a rare superior cerebellar artery giant, partially thrombosed aneurysm in which additional i.v. contrast-enhanced MRA sequences were comparable to digital subtraction angiography. It demonstrated not only the exact spatial resolution and correct anatomical relation but also the hemodynamics which were confirmed by intraoperative Doppler ultrasound. This report supports the feasibility and utility of i.v. contrast-enhanced MRA for posterior fossa giant cerebral aneurysm management. PMID- 9864735 TI - Expression of c-myc and c-fos and binding sites for estradiol and progesterone in human pituitary tumors. AB - We studied the concentration of mRNA from the oncogenes c-myc and c-fos in human pituitary adenomas by Northern blot hybridization (35 somatotrophinomas, 9 prolactinomas, 21 nonsecreting and 3 adrenocorticotrophinomas). The concentration of estrogens and progesterone receptors was also investigated. The levels of c myc and c-fos mRNA was higher in nonsecreting tumors which were generally the largest and had a higher percentage of recurrence after surgery than the other groups. High concentration of estrogen receptors was observed in tumors derived from cells which are normally the target of this hormone, mainly prolactinomas. They were also present in somatotrophic and nonsecreting adenomas, related to the presence of prolactin or gonadotrophin cells in these tumors. The presence of estrogen receptors indicates that the tumor cells maintain their differentiation and a good prognosis as is the case for prolactinomas. We did not find any relationship between estrogen receptors and the concentration of c-myc and c-fos oncogenes. Larger adenomas (mainly nonsecreting) had higher levels of c-myc and c fos mRNA than the other tumors and they had an important percentage of recurrence after surgery. It is clear that tumor size is related to the outcome after surgery and that nonsecreting adenomas are usually large because of the late diagnosis. However two large somatotrophinomas with extrasellar expansion also had overexpression of both oncogenes and both relapsed after surgery. PMID- 9864736 TI - Mobile fluoroscopic open stereotaxy for MRI-negative angiographic microlesions. AB - The approach to the deep-seated angiographic microlesion is often difficult, particularly when it is not demonstrated by computed tomography (CT) or magnetic resonance imaging (MRI). We have developed a method to localize these lesions for open stereotactic surgery employing mobile fluoroscopy. Prior to craniotomy, the patient's head is fixed in a stereotactic frame in a position optimal for the routine microscopic surgery. Following the injection of contrast media, the location of the lesion is marked on the fluoroscope monitor. Under fluoroscopic control, the scalp is marked using radiopaque pointer on each side of the patient's head so that the scalp marks and the target lesion overlap each other on the fluoroscope monitor. Thus the imaginary line connecting these scalp marks passes through the lesion. An additional pair of scalp marks is obtained by changing the projection angle of the fluoroscope. By simple calculation, the coordinates of the lesion are obtained as the nearest point to these two imaginary lines, each of which connects a pair of scalp marks. After craniotomy, the lesion is approached using an open stereotactic technique. The first patient had an aneurysm 1.5 mm in diameter that arose from the feeder of the arteriovenous malformation. The second patient had a small residual nidus of arteriovenous malformation 1.5 cm in diameter in the deep frontal lobe, not recognizable by CT or MRI because of artifacts from a previous surgery. Both patients were successfully operated by employing the present method. This method requires only a conventional stereotactic frame and a mobile fluoroscope, and provides simple and reliable localization of the small lesions recognizable only by cerebral angiography. PMID- 9864737 TI - Effects of mild (33 degrees C) and moderate (29 degrees C) hypothermia on cerebral blood flow and metabolism, lactate, and extracellular glutamate in experimental head injury. AB - The effects of mild (33 degrees C) and moderate (29 degrees C) hypothermia were investigated to determine which temperature was more effective against compression-induced cerebral ischemia. Eighteen cats were anesthetized. The animals were divided into three groups according to deep-brain temperature (control, 37 degrees C; mild hypothermia, 33 degrees C; and moderate hypothermia, 29 degrees C). Intracranial pressure (ICP) and cerebral blood flow (CBF) were monitored, the latter by hydrogen clearance. Arteriovenous oxygen difference (AVDO2) and cerebral venous oxygen saturation (ScvO2) were measured in blood samples from the superior sagittal sinus. The cerebral metabolic rate of oxygen (CMRO2) and the cerebral metabolic rate of lactate (CMR lactate) were calculated. Extracellular glutamate was measured by microdialysis. ICP was increased by inflation of an epidural balloon until CBF became zero, and this ischemia was maintained for 5 min, after which the balloon was quickly deflated. All parameters were recorded over 6 h. Evans blue was injected to examine vascular permeability changes. CBF was decreased by 56% by mild hypothermia and by 77% by moderate hypothermia. Mild hypothermia had a coupled metabolic suppression whereas moderate hypothermia significantly increased AVDO2 and decreased ScvO2, producing a low CBF/CMRO2 (relative ischemia). After balloon deflation, all three groups showed reactive hyperemia, which was significantly reduced by mild and moderate hypothermia. CBF then decreased to 50% of pre-inflation values and ScvO2 decreased (post-ischemic hypoperfusion). CBF/CMRO2, ScvO2, and AVDO2 did not differ significantly between the three groups. After balloon deflation, all three groups showed increased CMR lactate, which was significantly reduced by mild and moderate hypothermia. Extracellular glutamate increased in control animals (3.8 +/- 1.72 microM), an effect most effectively suppressed in the mild hypothermia group (1.0 +/- 0.46 microM). Damaged tissue volumes as indicated by Evans blue dye extravasation were 729 +/- 89 mm3 in control, 247 +/- 56 mm3 in mild hypothermia, and 267 +/- 35 mm3 in moderate hypothermia animals. These data suggest that mild hypothermia (33 degrees C) might be the optimal brain temperature to treat compression-related cerebral ischemia. PMID- 9864738 TI - Vascular compression of cranial nerves. I. History of the microvascular decompression operation. AB - The development of the microvascular decompression (MVD) operation is reviewed. It is stressed that a few innovative neurosurgeons discovered the role of vascular compression of cranial nerves V and VII in trigeminal neuralgia (TGN) and hemifacial spasm (HFS) and developed an operation, later to be known as the MVD operation. While the understanding of the pathophysiology of these disorders has improved, the surgical procedure has undergone little change since Gardner described the operation about 1960. PMID- 9864739 TI - Efficacy of the stump pressure ratio as a guide to the safety of permanent occlusion of the internal carotid artery. AB - To determine whether the absolute value for the stump pressure might be a useful index of symmetrical cerebral blood flow (CBF), and to examine correlations with the stump pressure ratio (initial mean stump pressure/preocclusion mean arterial pressure), fifty candidates for ICA injury or permanent occlusion were evaluated preoperatively. Each was continuously monitored for mean stump pressure and arterial pressure before, during (for a total of 20 min), and after balloon test occlusion. During the occlusion, CBF was measured by 99 m Tc-hexamethyl propyleneamine oxime (99 m Tc-HMPAO) single photon emission computed tomography (SPECT). The stump pressure and the stump pressure ratio were then compared with the results of 99 m Tc-HMPAO SPECT. Patients who failed to tolerate even brief periods of carotid occlusion and showed asymmetric decreases in CBF on SPECT were divided into high and moderate risk groups. Those with no significant changes in CBF on the occluded site formed the minimum risk group. Mean stump pressure was over 50 mmHg in 10 of a total of 25 patients in the high and moderate risk groups, and below 50 mmHg in 5 of the 25 patients in the minimum risk group. The stump pressure ratio did not exceed 56% in any but two patients in the high and moderate risk groups, and values were at least 60% in all patients of the minimum risk group. Decrease of CBF in two moderate risk group cases was localized in the posterior circulation. Difference in symmetrical CBF between the stump pressure ratio vs. the absolute value of mean stump pressure were statistically significant (p < 0.01, Fisher's Exact Test). Maintenance of a stump pressure ratio of 60% or more during test occlusion may be a more useful index for a good collateral circulation than any absolute value for mean stump pressure. PMID- 9864740 TI - Direct ethanol injection for skull metastasis from hepatocellular carcinoma. The techniques and consequences of a therapeutic trial. AB - The frequency of skull metastasis from hepatocellular carcinoma (HCC) is increasing together with the recent progress in the management of the primary lesion. Cases are often complicated with poor general conditions or metastasis to the other organs, and not readily indicated for surgery. A direct injection of ethanol to the lesion could be one of the therapeutic options to cope with this complicated situation. To evaluate the feasibility of this technique, we planned a therapeutic trial in a patient with HCC associated with lumbar and skull metastasis, the latter metastasis repeated twice during the past one year. A total of 10 ml of ethanol was injected into the skull metastasis percutaneously under ultrasound (US) guidance. US guidance was very useful in determining the sites of injections and the distribution of ethanol as well as monitoring the blood flow within the tumor vessels. The patient transiently complained of local pain at the injection sites, but there were no other adverse effects. Four days after the injection, the lesion was resected by surgery, which confirmed the pathologic diagnosis as well as the nearly-total necrosis of the tumor. This technique is simple, safe and repeatable with low cost. The technical details and the histologic effects are described. PMID- 9864741 TI - Prolonged survival of rats with intracranial C6 gliomas by treatment with TGF beta antisense gene. AB - Using an intracranial rat C6 glioma model, we tested the hypothesis that gene modification of glioma cells to block the expression of the immunosuppressive cytokine TGF-beta (transforming growth factor beta) may enhance anti-tumor immune responses and thereby prolong survival of tumor-bearing animals. The cDNA for simian TGF-beta 2 was ligated in antisense orientation into the episomal plasmid mammalian expression vector pCEP-4. This TGF-beta-antisense vector was transfected into C6 glioma cells by standard electroporation techniques. PCR was used to determine that the rat C6 clones were successfully transfected with the antisense-TGF beta construct. Twenty-nine adult female Wistar rats harboring 7 day-old intracranial C6 tumors were then subcutaneously injected with either saline (n = 9), unmodified C6 glioma cells (n = 10), or TGF-beta-antisense modified C6 cells (n = 10). Animals were followed for survival, and Fisher's exact method was used to interpret the significance of difference between experimental groups. The survival of tumor-bearing rats injected with TGF-beta antisense-modified C6 cells was significantly prolonged, relative to the survival of rats receiving injections of saline or unmodified C6 cells alone. Six of the ten (60%) TGF-beta-antisense treated animals survived for 12 weeks, whereas none of the nine (0%) animals treated with saline and none of ten (0%) of those treated with C6 cells alone survived past 5 weeks. These results indicate that the genetic inhibition of immunosuppressive cytokines (such as TGF-beta) may reverse the phenotypic immunosuppression caused by such factors, and thereby prolong the survival of C6 tumor-bearing animals. Future investigations using cytokine gene modifications in other brain tumor models are warranted. PMID- 9864742 TI - Hypervascular acoustic neuroma. AB - The authors present a patient with a large acoustic neuroma that exhibited an unusual vascular architecture. Magnetic resonance imaging demonstrated multiple flow voids in and around the mass. At surgery, intra- and extratumoral vascularity was arterialized due to luxurious shunting. Two attempts at removal produced severe engorgement and pulsatility of the surrounding brain, dramatically narrowing the operative exposure. Piecemeal resection of the tumor seemed to redistribute blood flow resulting in engorgement of the surrounding brain, analogous to perfusion breakthrough following AVM resection. Treatment strategies similar to those used for AVM resection may be indicated when encountering a hypervascular tumor. PMID- 9864743 TI - Scalp localization using a simple algorithm. AB - We have developed a simple algorithm for scalp localization for craniotomy. Using a series of CT/MRI data, the point on the scalp closest to the center of the lesion (T) is determined by our program. A plane (plane R) is defined by three points: bilateral auditory meati and T. Point S is the intersection of three planes: the plane R, the scalp surface and the midsagittal plane. The distance from either S or the ipsilateral external auditory meatus to T is measured along the scalp surface on the plane R. The distance from the nasion to S along the scalp surface on the midsagittal plane is also measured. To determine the craniotomy site, these distances are measured directly on a patient's scalp in the operating room. This simple and accurate method for scalp localization could be used by installing our program in conventional CT/MR scanners. PMID- 9864744 TI - Motor speech impairment following traumatic brain injury in childhood: a physiological and perceptual analysis of one case. AB - The physiological and perceptual characteristics of persistent dysarthria exhibited by a 14 year-old boy, following a severe traumatic brain injury (TBI) were investigated. The subject's speech and motor speech mechanism were comprehensively evaluated both perceptually and physiologically, and the findings were compared with those of a non-neurologically impaired control subject, matched for age and sex, and a number of control groups from previous studies. Overall, the assessments indicated that the major motor speech deficits demonstrated by the subject included severely reduced tongue function, and moderately impaired lip, laryngeal, and velopharyngeal function. Respiratory function was found to be mildly impaired. Perceptual assessments indicated that the subject's speech was severely impaired in relation to rate, pitch variation, and consonant precision, with a moderate impairment in overall intelligibility. The effects of a severe TBI on the functioning of the child's motor speech mechanism were discussed. The clinical implications for the assessment and treatment of dysarthria in childhood following severe TBI were highlighted. PMID- 9864745 TI - Rehabilitation and orthotic management of congenital and acquired meromelia in the lower limbs of children. AB - Children with limb deficiencies or amputations of the lower extremities should be enabled to stand or walk according to their state of development, because standing and walking are among the most important preconditions for the best possible integration. Supplying them with a prosthesis, orthosis and orthoprosthesis is therefore indispensable for rehabilitation. While a prosthesis replaces parts of the extremities, an orthosis stabilizes the existing extremity. Orthoprostheses compensate longitudinal malformations, have a supporting effect, allow growth to be controlled and compensate for shortening. Just as important as the remedy is early medical treatment. The medical team taking care of the patient works out a treatment plan where responsible cooperation with the parents is of utmost importance. The focus of all efforts is, of course, the handicapped child. Examples are used to demonstrate the course of successful rehabilitation of children and adolescents with damaged limbs. Equally the psychological situation of the parents and child is taken into consideration. PMID- 9864746 TI - They cannot sit properly or move around: seating and mobility during treatment for developmental dysplasia of the hip in children. AB - Children diagnosed with Developmental Dysplasia of the Hip (0.2% of live births) are often treated by splintage to hold the head of the femur in the acetabulum during early joint development. Whilst clinically effective, this can create difficulties for the parents in handling the child and affects the mobility of the family, which subsequently creates emotional and social difficulty resulting from the disruption of the family routine. To identify these problems and their order of priority, a survey of 113 recently affected families was carried out in England and Northern Ireland. Parents identified mobility, emotional and social problems. Splintage size and shape was the fundamental problem from which the other difficulties arose. Solutions to the basic difficulties of transporting and seating a child in splintage would largely alleviate the feelings of frustration felt by the families and enable more normal activities of daily living. PMID- 9864748 TI - Hot topics in motor control and learning: introduction. The dynamic systems approach to motor control and learning: promises, potential limitations, and future directions. PMID- 9864747 TI - Postural adaptation in nemaline myopathy: observations in two siblings. AB - Two siblings with nemaline myopathy were analysed. There was a general muscle weakness and lack of muscle strength that was more obvious in the proximal muscles. One could walk independently and one with minimal support. Patients developed a serial of postural adaptations. No correlation between muscle strength and functional level was found. PMID- 9864749 TI - A dynamic systems perspective to perception and action. PMID- 9864750 TI - An alternative view of dynamical systems concepts in motor control and learning. PMID- 9864751 TI - On the dynamics of motor learning research. AB - In this paper I have attempted to conceptualize the dynamical pattern view of learning (Zanone & Kelso, 1992, 1994, 1997) in terms of some historical similarities to the dynamic pattern perspective, unique contributions, and potential problems to be addressed in future research and theory. For researchers interested in motor learning issues, all three of these points can be viewed only as positive advances to the field of study. PMID- 9864752 TI - The reliability and validity of a chair sit-and-reach test as a measure of hamstring flexibility in older adults. AB - The purpose of this study was to examine the test-retest reliability and the criterion validity of a newly developed chair sit-and-reach (CSR) test as a measure of hamstring flexibility in older adults CSR performance was also compared to sit-and-reach (SR) and back-saver sit-and-reach (BSR) measures of hamstring flexibility. To estimate reliability, 76 men and women (M age = 70.5 years) performed the CSR on 2 different days, 2-5 days apart. In the validity phase of the study, scores of 80 men and women (M age = 74.2 years) were obtained on three field test measures of hamstring flexibility (CSR, SR, and BSR) and on a criterion test (goniometer measurement of a passive straight-leg raise). Results indicate that the CSR has good intraclass test-retest reliability (R = .92 for men; r = .96 for women), and has a moderate-to-good relationship with the criterion measure (r = .76 for men; r = .81 for women). The criterion validity of the CSR for the male and female participants is comparable to that of the SR (r = .74 and r = .71, respectively) and BSR (r = .70 and r = .71, respectively). Results indicate that the CSR test produces reasonably accurate and stable measures of hamstring flexibility. In addition, it appears that the CSR is a safe and socially acceptable alternative to traditional floor sit-and-reach tests as a measure of hamstring flexibility in older adults. PMID- 9864754 TI - Mass and velocity: control parameters for throwing patterns. AB - The purpose of this study was to determine if change in segmental mass and increases in throwing velocity act as control parameters to alter throwing patterns. Twenty participants were categorized into four levels of throwing pattern. Each participant was required to make 10 throws at various velocities within 8 conditions. Conditions resulted from combinations of altered mass of the arm, forearm, and hand. Quantitative and qualitative analyses indicated that changes in segmental mass and release velocity either increased or decreased levels of throwing pattern depending on the initial throwing level. It was concluded that mass and velocity may be control parameters which instigate changes in throwing patterns. PMID- 9864753 TI - Bilateral reaching to asymmetrical targets: muscle and joint dynamic interlimb adaptations. AB - A combined analysis of time, electromyographic, and joint torque measures was used to explore the force control processes underlying the dissociation of arm reaching movements performed bilaterally to targets of varying amplitude. Limb movements appeared closely coupled at movement initiation, which was confirmed by a strong tendency of the agonist muscles to remain synchronized despite any interlimb asymmetry in final target distance. On the other hand, interlimb decoupling occurred later as a result of the difference in the antagonists' timing of activation between the limbs. The partitioning of the net joint torque revealed that muscle activity is regulated in response to the intersegmental dynamics of the limb. It is proposed that spatial decoupling of asymmetrical movements becomes possible through postinitiation feedback processes which regulate muscle recruitment phenomena. PMID- 9864755 TI - Skill and tactical development during a sport education season. AB - The purpose of this study was to examine the development of skill competence and tactical sophistication during a games unit conducted following the features of sport education provided by Siedentop (1994). One team of six players was followed through a 30-lesson season of "Ultimate." Using Grehaigne, Godbout, and Bouthier's (1997) formula for an efficiency index, together with a number of other measures of skill and tactical competence, it was determined that these students made significant improvements in selection and execution dimensions of the game. Furthermore, lower skilled students did not feel marginalized within their teams and believed they had equal opportunities for improvement. The key reasons for developing competence in this setting were the sufficient length of the season to allow for significant practice opportunities and the consistent team membership throughout season, which allowed all players to develop a sense of usefulness. This curriculum model provides one way students can develop skills through a system of game playing, provided sport education units are structured to prevent the more skillful players from dominating the games. PMID- 9864756 TI - Observational learning and the fearful child: influence of peer models on swimming skill performance and psychological responses. AB - This study examined the role of peer mastery and coping models on children's swimming skills, fear, and self-efficacy. Children (N = 24; M age = 6.2 years), who were identified as fearful of the water, were matched to control, peer mastery, or peer-coping model conditions. Day 1 included a preintervention assessment. Days 2-4 included exposure to model conditions followed by a 20-min swimming lesson, Day 5 consisted of postintervention assessments, and a follow-up test was conducted 4 days later. Data were analyzed in a series of 3 x 3 (Model Type x Assessment Period) repeated measures analyses of variance on the dependent variables. Results revealed differences between modeling and control groups at postintervention and follow-up, but the small sample size and large within-group variability compromised many statistically significant findings. Calculation of effect sizes indicated moderate-to-large pre- to posintervention differences between control and modeling groups on skill, self-efficacy, and fear of swimming. These findings suggest that a modeling intervention combined with swimming lessons is a more effective behavior change agent for fearful children than swimming lessons alone. PMID- 9864757 TI - The validity of the Tritrac-R3D activity monitor for the assessment of physical activity: II. Temporal relationships among objective assessments. PMID- 9864758 TI - Analysis of event-related potentials to filmed action situations. PMID- 9864759 TI - Attentional demands and the organization of reaching movements in rock climbing. PMID- 9864760 TI - Acute muscle stretching inhibits maximal strength performance. PMID- 9864761 TI - A test of the reliability and factorial validity of the Greek version of the CSAI 2. PMID- 9864763 TI - [Patients' needs and quality of care]. PMID- 9864762 TI - Task structures, feedback to individual students, and student skill level in physical education. PMID- 9864764 TI - [Relations between blood pressure at 3-4 years of age and body mass at birth: a population-based study]. AB - BACKGROUND: To study the association between blood pressure of 3-4 year-old children and birth corpulence estimated from birth ponderal index (body weight/length3), a school based, cross sectional survey was conducted in the Rhone department. METHODS: 1296 randomly selected children were examined at the time of a school medical visit. Physicians abstracted information on pregnancy and measurements at birth from the Child Health Handbook and from parents interview. Systolic and diastolic blood pressure measurements were carried out by using an automated oscillometric device. RESULTS: Multiple linear regression analyses revealed an inverse association between systolic blood pressure and birth ponderal index (converted into Z-score adapted to gestational age), this was obtained after adjustment for current length and for current body mass index (body weight/length2--converted into Z-score), for child emotion and for linked study factors. Such a model provided an assessment of regression coefficient: 0.67 mmHg/birth ponderal index Z-score unit (95% confidence interval: -1.24 to 0.11). So, estimation of systolic blood pressure difference between two 3-4 year old children, one at -2 SD birth ponderal index and another at +2 SD, was obtained: 2.6 mmHg. No evidence of an association between diastolic blood pressure and birth ponderal index was observed. CONCLUSION: In this large population with a limited range of age, our results show that, at age 3-4 years, systolic blood pressure is higher with lower birth ponderal index, suggesting that thinness at birth might be a risk factor of hypertension in adulthood. PMID- 9864765 TI - [Access to social coverage of uninsured patients attending a hospital clinic: a historical cohort study at the Baudelaire outpatient clinic in Paris]. AB - BACKGROUND: In France health insurance coverage is universal (see note at the end of the text), nevertheless some people remain uninsured. In this high-risk population, the lack of insurance coverage contributes to the aggravation of health, by reducing access to medical care. In 1992, the Baudelaire consultation was incorporated into the outpatient clinic of Saint-Antoine hospital (Paris, France), to provide the uninsured with the same access as any other patient--but free of charge--to medical care. Social care was also provided in particular by assisting the uninsured in applying for insurance coverage. Our objectives were to quantify the delay in obtaining insurance coverage and to study whether the sociodemographic characteristics of these patients were associated with inequalities in terms of delays. METHODS: All patients attending the consultation for the first time in 1994 were included (n = 623). Because of differences linked to the French social security system, analysis was performed into two groups according to the existence of a prior insurance coverage. Delay in obtaining or recovering insurance coverage was considered as the key variable. The socio demographic factors linked to the rates of access to insurance coverage were determined using Cox proportional hazards regression models. We also examined the factors linked with the existence of a prior insurance coverage by logistic regression modeling. RESULTS: Within one year 96% of the patients who had had insurance coverage in the past, and 63% of the patients who had not, were insured. No factor, whether nationality, educational level, socio-professional category, family situation, type of housing, made of income was found to be linked with obtaining or recovering insurance coverage. However, nearly all these factors were related with the existence of prior insurance coverage. CONCLUSIONS: Our approach of systematically providing social care allows 70% of uninsured patients to obtain insurance coverage within one year. This approach probably contributes to an improvement by facilitating access to mainstream health care. Moreover, no difference in delay in obtaining insurance coverage was found associated with sociodemographic characteristics. PMID- 9864766 TI - [Internal consistency, factorial validity and discriminant validity of the French version of the psychological demands and decision latitude scales of the Karasek "Job Content Questionnaire"]. AB - BACKGROUND: The job demands-control model developed by Karasek has greatly influenced research on psychosocial factors at work and health. Validity of the English version of the psychological demands and decision latitude scales is documented. Psychometric qualities of the French version are investigated here in a representative sample of the general population, including blue-collars and white-collars. METHODS: The French translation of the psychological demand and decision latitude scales was administered by interview in a representative sample of the Quebec working population (N = 1,110). Internal consistency and factorial validity of the instrument were studied among white-collars and blue-collars separately. Discriminant validity was assessed for the whole population. RESULTS: Cronbach alpha coefficients, varying between 0.68 and 0.85, support the internal consistency of the scales. Demographic distribution of the scales and intercorrelations were consistent with the English version. Results of the factor analysis were consistent with the two dimensions expected from the theory. Mean scale scores and variations in the prevalence of high psychological demands combined with low decision latitude by age, sex, education, and job category support the discriminant validity of the instrument. CONCLUSIONS: Results support internal consistency, factorial validity, and discriminant validity of the French version of the psychological demands and decision latitude scales of the Karasek "Job Content Questionnaire" for white-collars and for blue-collars of the general population. PMID- 9864767 TI - [Measurement of anxiety state in women: a short-form scale]. AB - BACKGROUND: Clinical epidemiology and psychosocial studies often take into account the measurement of anxiety. Our objective was to develop and validate a short form scale of anxiety state from the Spielberger's State Trait Anxiety Inventory (STAI-Y), one of the most widely used scales. METHODS: This study was carried out on French samples of women with different levels of anxiety. The first step was to select from the original 20-items scale, the items susceptible to compose two short forms scales: an 8-items (18-STAI) and a 5-items (15-STAI) one. It was carried out on patients attending cancer genetic clinics (n1 = 160, n2 = 41). The second step was carried out on 3 other samples of women (n3 = 150, n4 = 167, n5 = 105) and measured the psychometric characteristics of the short form scales obtained previously. RESULTS: The correlation coefficients between the short-forms and the complete 20-items form were very high (> 0.90) in particular for the 18-STAI (r > 0.95). Both short forms are sensitive to change. The internal consistency measured by Cronbach-alpha was comparable to the original scale. The results obtained with the 18 scale and another 10 items Spielberger's short-form scale are comparable and better than those with the 15 scale. CONCLUSION: The 18 French short-form scale, balanced with positive and negative items, can be recommended for the measurement of state anxiety when the complete form cannot be used. PMID- 9864768 TI - Should condom use be promoted for contraception to prevent transmission of sexual transmitted diseases and AIDS? ACSF Group. French National Survey of Sexual behavior. AB - BACKGROUND: This study is an exploration of the relationship between the use of oral contraceptive or intrauterine device and condom use. The results should help us better understand how promoting condoms for contraception as compared to sexually transmitted disease prophylaxis might be an effective way of increasing their use in certain populations. METHODS: Women not involved in stable relationships were selected from the French National Survey on Sexual Behaviour using multi-stage probability sampling. We compared frequency of condom use at the last sexual intercourse by women who were using prescribed contraception with that by women who did not. Odds ratios (OR) were adjusted for age, duration of the relationship, education, type of partner and number of partners in the last 12 months. RESULTS: Overall, no difference was found in the frequency of condom use during the last sexual intercourse by women who were using prescribed contraception compared with women who were not. A more frequent use of condoms tended to be associated with the absence of contraception in relationships of less than three months' duration (adjusted OR: 4.5; 95% CI: 0.7-33.3) and for women of 25 to 34 years of age (adjusted OR: 2.4; 95% CI: 0.8-7.1). CONCLUSION: Many factors other than contraceptive practice influence the use of condoms. Nevertheless, our results suggest that reproductive issue should be taken into account in specific subgroups of the population when developing policies to prevent sexual transmission of HIV. PMID- 9864769 TI - [Psychosocial factors at work and cardiovascular diseases: contribution of the Effort-Reward Imbalance model]. AB - The effect of psychosocial factors at work on health, especially cardiovascular health, has given rise to growing concern in occupational epidemiology over the last few years. Two theoretical models, Karasek's model and the Effort-Reward Imbalance model, have been developed to evaluate psychosocial factors at work within specific conceptual frameworks in an attempt to take into account the serious methodological difficulties inherent in the evaluation of such factors. Karasek's model, the most widely used model, measures three factors: psychological demands, decision latitude and social support at work. Many studies have shown the predictive effects of these factors on cardiovascular diseases independently of well-known cardiovascular risk factors. More recently, the Effort-Reward Imbalance model takes into account the role of individual coping characteristics which was neglected in the Karasek model. The effort-reward imbalance model focuses on the reciprocity of exchange in occupational life where high-cost/low-gain conditions are considered particularly stressful. Three dimensions of rewards are distinguished: money, esteem and gratifications in terms of promotion prospects and job security. Some studies already support that high-effort/low reward-conditions are predictive of cardiovascular diseases. PMID- 9864770 TI - [Appropriateness of medical procedures: measurements and relations with needs and access to health care]. AB - Efforts to reduce cost expenditures and improve quality of care have raised some concern over the recent years as to the appropriateness of medical procedures. However, the relationships between the appropriateness of medical procedures, the need for such procedures and access to health care have not been described in detail. The appropriateness of medical procedures is usually measured on the basis of procedures actually performed. We present a critical appraisal of this type of measurement. From a population point of view, measurement of appropriateness does not actually reflect simply those procedures which have been performed, but rather involves three parameters: needs for a particular procedure, access to the procedure by those who need it, and non performance of the procedure among those who do not need it. This analysis revealed three consequences. First, there is an inverse relationship between access to a procedure among subjects who need the procedure and the appropriateness of such a procedure among those who do not need it. Secondly, comparisons of appropriateness rates between individual facilities or between regions must take into account the need for this procedure in the general population. Finally, it is possible to observe similar rates of procedure performance between regions or social groups despite major regional or social differences in appropriateness and access to health care. PMID- 9864771 TI - [Detection of cancer, sensitivity of the test and sensitivity of the screening program]. AB - In assessment of screening for cancer, no distinction is usually made between the sensitivity of the screening test (St) and the sensitivity of the screening program (Sp). This paper was aimed to distinguish meaning, method for assessment and interest for each of them, and to determine their relationship. Sensitivity of the screening program can be directly assessed with data from on-going trials whilst assessment of sensitivity of screening test requires modelisation techniques, especially for assessing the mean duration of the preclinical phase of cancer. Assuming an exponential distribution of this duration, lambda as the time parameter, a mathematical relation between St and Sp is suggested as follows: [formula: see text] with r being the interval between two screening tests. The implementation of this equation with data from a mass-screening program for colorectal cancer in the department of Calvados allowed us to investigate the influence of the mean preclinical phase and the interval between two screening tests on the value of the sensitivity of the screening procedure. Such a modelisation could be useful in the development of a rational screening policy. PMID- 9864772 TI - [Taking into account patients' expectations in the improvement of quality of health care: results of a survey of 500 hospitalized patients]. PMID- 9864773 TI - [AIDS. Account of the 12th world conference on AIDS Geneva, 28 June-3 July 1998]. PMID- 9864774 TI - [Amyloidosis. Account of the 8th international symposium on amyloidosis Rochester Minnesota, United States, 7-11 August 1998]. PMID- 9864775 TI - [Heparin-induced thrombopenias: new treatments]. PMID- 9864776 TI - [End-stage renal failure following parathyroidectomy for advanced primary hyperparathyroidism]. AB - PURPOSE: We report two cases of long-standing, advanced primary hyperparathyroidism with renal failure. After surgery the two patients presented with marked hypocalcemia and deterioration of renal function which required dialysis 2 and 3 months after parathyroidectomy. These two cases lead us to consider that renal function should be studied before and after surgery in patients operated on for primary hyperparathyroidism. METHODS: In 19 patients operated on for hyperparathyroidism with few symptoms and without renal failure or with very moderate renal failure, blood creatinine and creatinine clearance were measured before (T1) and 1 year (T12) parathyroidectomy. RESULTS: No significant overall changes in the following parameters were observed: blood creatinine T1 71 +/- 19 mumol/L, T12 82 +/- 20 mumol/L, CrCl T1 72 +/- 13 mL/min, CrCl T12 70 +/- 19 mL/min. However, in the patient with greatest deterioration of renal function, CrCl decreased from 45 mL/min at T0 to 33 mL/min at T12. CONCLUSION: A review of the literature shows that in certain cases, renal insufficiency present before cervicotomy may worsen after surgery even if hypercalcemia is corrected. The mechanism is still unclear. A sharp decrease in parathormonemia and parathyroid hypertensive factor may play a role via intrarenal hemodynamic changes. PMID- 9864777 TI - [Comparison of cases of isolated pulmonary tuberculosis with cases of other localizations of tuberculosis in the course of an active surveillance (Gironde, 1995-1996)]. AB - PURPOSE: Data collected during the years 1995 and 1996 in the course of an epidemiological survey of tuberculosis in Gironde allowed comparison of pulmonary tuberculosis with extrapulmonary localizations, evaluation of the importance of each localization and highlighting of potential risk factors. METHODS: Patients living in Gironde who had evidence of either clinical, radiological or bacteriological expression of tuberculosis were included in the survey. Statistical comparisons were done using either Pearson's Chi 2 or Fisher's exact test. RESULTS: The survey included 292 cases subdivided into 183 cases of pulmonary tuberculosis (63%) and 109 cases in which another localization had been diagnosed (37%). Extrapulmonary localizations that were the most often encountered either alone or in association with pulmonary localization were the following: lymphadenopathy (32%), pleural (28%), genito-urinary (12%) and osteo articular localizations (7%). The survey showed that patients in whom tuberculosis localization was extra-pulmonary were more frequently under 20 years of age or over 60 years of age (P < 0.04). These patients also presented more often with HIV-infection (P < 0.02). CONCLUSION: Extrapulmonary localizations of tuberculosis should be systematically investigated in young and elderly patients as well as in HIV-infected patients. PMID- 9864778 TI - [Role of corticosteroid therapy in non-malignant diseases]. AB - INTRODUCTION: As short-term corticosteroid therapy is widely used in clinical practice, it is important to determine its precise indications and limits of use. CURRENT KNOWLEDGE AND KEY POINTS: Duration of short-term corticosteroid therapy is arbitrarily considered to be up to 21 days. Anti-inflammatory, antiproliferative or analgesic actions represent the main pharmacological features of steroids. They are related to the interactions of steroids with cytokines and immune cells. Results of randomized double-blind and uncontrolled clinical studies were included in this review. Furthermore, clearly demonstrated results that were obtained more particularly in neurology, otorhinolaryngology, pneumology, infectious diseases, rheumatologic and traumatic processes are summarized. FUTURE PROSPECTS AND PROJECTS: Indications for short-term corticosteroid therapy are well established. However, further clinical studies are required, as current prescription of corticosteroid is still empirical in the management of most diseases. PMID- 9864779 TI - [Malignant pleural effusions: recourse to early use of talc]. AB - INTRODUCTION: Our purpose was to assess the efficacy, permanence and safety of thoracoscopic talc poudrage (TTP) for pleurodesis in malignant effusions. We report the follow-up of 360 patients who received TTP in two centers in Marseille (France). CURRENT KNOWLEDGE AND KEY POINTS: Eighty-eight patients presented with mesothelioma and 272 had pleural metastasis. The mean follow-up time was 12 months (range: 2-120). Out of the 327 patients whose response could be evaluated, 90.2% had a successful pleurodesis at 1 month, and 82.1% had a life-long pleural symphysis. Adverse effects included one death 3 days after the procedure in an end-stage patient, fever (9.8%), infection of the parietal scar (2.5%) and pulmonary infection (0.8%). FUTURE PROSPECTS AND PROJECTS: TTP is an effective and safe method of life-long pleurodesis. It should be performed early on in the history of malignant effusions to avoid failures of the technic, mainly linked to trapped lung and to the general condition of patients. PMID- 9864780 TI - [Heparin-induced thrombopenia: rapid regression of thrombopenia and thrombosis after plasma exchange. Case report]. AB - INTRODUCTION: Heparin-induced thrombocytopenia with thrombosis is a rare but severe adverse reaction to heparin therapy, whose management is difficult. After heparin withdrawal, the initiation of an alternative anticoagulant therapy, such as recombinant hirudin or danaparoid, is strongly recommended before vitamin K antagonists are effective. Several reports of the efficacity of plasma exchanges in patients with life-threatening thrombosis have been made. EXEGESIS: We report on a patient with severe aortic thrombosis related to heparin therapy in whom a unique plasma exchange resulted in both dramatic improvement in the platelet count and marked reduction of the thrombosis. CONCLUSION: This case provides further evidence that plasma exchanges can be useful in the management of heparin induced thrombocytopenia with thrombosis. They are rapidly efficient and can be used before heparin alternative treatment is effective. PMID- 9864781 TI - [Pseudo-Glanzmann thrombasthenia in the course of autoimmune thrombocytopenic purpura]. AB - INTRODUCTION: Auto-immune thrombocytopenic purpura is associated with platelet anti-glycoprotein antibodies, particularly with anti-GPIIb/IIIa complex. Persistence of these antibodies sometimes leads to acquired auto-immune thrombopathy. EXEGESIS: We report the case of a woman treated by splenectomy for auto-immune thrombocytopenic purpura, who developed 5 years later an ecchymotic syndrome despite normal platelet count. High bleeding time and platelet aggregation defect in vitro were evidenced. Following the initial thrombocytopenia, anti-glycoproteins GPIIb/IIIa with lupus anticoagulant and benign monoclonal gammapathy were noticed. Platelet controls showed that hypoaggregant activity was secondary to the persistence of anti-GPIIb/IIa antibodies. CONCLUSION: This acquired auto-immune thrombopathy simulating Glanzmann's thrombasthenia was secondary to the persistence of platelet anti glycoproteins GPIIb/IIIa. PMID- 9864782 TI - [Pulmonary textiloma revealed by hemoptysis 12 years after thoracotomy]. AB - INTRODUCTION: Pulmonary textiloma or retained surgical sponge is rare but can have detrimental consequences. Its diagnosis is difficult, even when using computerized tomography (CT) scan and is always a medical and legal problem. EXEGESIS: We report the case of a 46-year-old man with a history of pulmonary tuberculosis diagnosed by thoracotomy in 1986, who was admitted to hospital for hemoptysis. CT scan showed the existence of a pulmonary lesion. Clinical, radiological and fiberoptic endoscopy features mimicked an abscess or bronchiectasis. The patient underwent left inferior lobectomy. Anatomical findings led to the diagnosis of textiloma. CONCLUSION: This case shows the difficult in diagnosing pulmonary textiloma, even when using CT scan. Textiloma should be considered when an atypical pulmonary mass is found in a patient with past history of thoracotomy. PMID- 9864783 TI - [Diabetes insipidus revealing primary malignant non-Hodgkin's lymphoma of bone]. AB - INTRODUCTION: Diabetes insipidus is a very rare, occasionally misleading, revealing symptom of non Hodgkin's malignant lymphoma. We report a case of this uncommon association and a review of the literature is done. EXEGESIS: Soon after a transient attack of diplopia due to a 6th cranial nerve palsy, idiopathic cerebral diabetes insipidus was diagnosed in a 32-year-old immunocompetent man. As anterior hypopituitarism was absent, the patient was discharged and treated with desmopressin acetate. Six months later, recurrence of diploplia together with raised erythrocyte sedimentation rate was observed and B cell lymphoma was observed in the iliac bone and skull, with extension into the right cavernous sinus. Both tumoral lesions led to important uptake of gallium-67. After a 26 month follow-up following intensive chemotherapy including autologous peripheral stem cell transplantation and local radiotherapy, lymphoma symptoms had disappeared, but the patient's condition had only partially improved with regard to diabetes insipidus. The association of lymphoma and diabetes insipidus which has only been reported in 14 cases is discussed. CONCLUSION: Since endocrine symptoms are often preceding hematological diagnosis, malignant lymphomas should systematically be viewed as potential etiology of diabetes insipidus. Magnetic resonance imaging which is more sensitive than computerized tomography in revealing brain lymphomas is mandatory. Gallium scan imaging might help guide the diagnosis in some instances. PMID- 9864784 TI - [A deformed hand]. PMID- 9864785 TI - [Splenic volvulus or the syndrome of the wandering spleen]. PMID- 9864786 TI - [Erythema nodosum revealing chronic myelomonocytic leukemia: two cases]. PMID- 9864787 TI - [Splenic lymphoma with villous lymphocytes and Waldenstrom disease: diagnostic difficulties by way of a case report]. PMID- 9864788 TI - [Eosinophilic ileocolitis: two case reports]. PMID- 9864789 TI - [Asthma and infection: risk or prevention?]. AB - Upper respiratory tract infections and wheezing illnesses occur frequently in early childhood. Most viral infections associated with wheeze in infancy are attributable to respiratory syncitial virus, whereas in older children rhinovirus, influenza and parainfluenza virus prevail. The detection rate of viruses has increased since PCR techniques have been used. A British study for example showed that in 80% of episodes in asthmatic children aged 9-11 years viruses could be detected in nasal aspirates which had been taken within 4 days of the wheezing attack. In most cases rhinovirus was identified. Two major hypotheses have been proposed to explain the association between respiratory tract infections and subsequent respiratory abnormalities. One hypothesis states that viral infections early in life damage the growing lung or alter host immune regulation. The second hypothesis holds that respiratory infections are more severe in infants and children with some underlying predisposition. These notions await further clarification. Early childhood bacterial and viral infections may, however, also be associated with a reduced risk of developing atopic sensitization or allergic conditions, as the results of several recent studies suggest. PMID- 9864790 TI - [Knowledge, attitude and reservations of medical students about organ transplantation: results of a survey during the first year of study]. AB - AIM: Establish a list of first year medical students' attitudes, doubts, and knowledge in the fields of organ transplantation and donation. METHOD: Anonymized questionnaire handed out to students during class lectures. RESULTS: 183 questionnaires were distributed and 117 returned (participation: 64%). The average age of the students was 21.6 +/- 2.7 years (range 18 to 38 years); the sample included 71 women (60.7%) and 48 men (39.3%). Only 2 students (2%) were not interested in the subject of organ donation. The students knew very little of the legal aspects of organ donation and 1/4 of them thought there was even a Federal law regarding organ transplantation. When asked if they knew whether a law existed in the Canton of Berne, 44% replied yes, but only 24 (20%) knew that this is contradictory. There was no gender difference in the answers to these question. From 57 students (48%) 246 individual comments on doubts and concerns were analyzed. In this respect, the students mainly questioned whether the donor was truly dead when donation took place (n = 48), if illegal transplantation could be eliminated (n = 44) and if transplantation was truly necessary (n = 43). Some also mentioned religious/ethical doubts (n = 42). In regard to organ donation by a living individual, 27 students were concerned about the health of this donor. 20 students had doubts regarding the pressure possibly applied by family members and friends and as many voiced doubts in regard to premature diagnosis of brain death of potential donors. Only 2 students were concerned about the post-mortem presentation. 45 students (48%) indicated discomfort with the donation of certain organs. They ranked the kidney as the first organ to donate, followed by the pancreas, heart, cornea, intestine, lung and liver. CONCLUSION: The interest in organ donation and transplantation is already strong in fist year medical students in the pre-clinical stage. However, differences from lay public are not readably detectable at this stage of medical training. Adequate information could influence future physicians in their mediatory role. PMID- 9864791 TI - [A patient with rapidly progressing renal failure, florid syphilis and positive HIV serology]. AB - We describe the frist reported case in Switzerland of HIV-associated nephropathy (HIVAN). HIVAN shows a typical combination of clinical findings: black race, proteinuria, large hyper-echogenic kidneys, normal blood pressure, positive HIV serology and no autoantibodies. The histologic findings are typical: focal segmental glomerulosclerosis of the collapsing variant, often with marked interstitial nephritis. The disease normally appears before AIDS symptoms develop and follows a very aggressive course to end-stage renal disease. Therapy consists of a combination of nucleoside reverse transcriptase and proteinase inhibitors, ACE inhibitors, and possibly steroids. In end-stage renal disease patients can be managed by haemodialysis, continuous ambulatory peritoneal dialysis (CAPD) or kidney transplantation. PMID- 9864792 TI - [Inguinal hernia 1998--an assessment of current status]. AB - Hernia surgery has considerably changed in recent years. In the era of minimal invasive surgery classical Shouldice repair has become old-fashioned and is increasingly replaced by tension-free techniques using synthetic mesh material. Currently, Shouldice repair remains the treatment of choice in young patients with small primary hernia. Lichtenstein hernioplasty is indicated in young patients with large hernias, and in those over 35 years of age for any size of hernias. Endoscopic operations are restricted to bilateral primary hernias and recurrent hernias. In future, when used on the basis of a reasonable strategy, the variety of operative procedures offers a chance to improve the results of hernia surgery. Further studies are needed to demonstrate which hernia strategy is most reliable in terms of cost-effectiveness, patient comfort, complication and recurrence rate. PMID- 9864793 TI - [Successful surgical management of dementia symptoms]. PMID- 9864794 TI - [Immunological aspects of diabetic angiopathies pathogenesis]. PMID- 9864795 TI - [Comparative study of peripheral circulation, lipid metabolism and hemocoagulation in families with atherosclerosis and diabetes mellitus]. AB - AIM: The study of peripheral circulation (PC) in patients with atherosclerosis (AS), diabetes mellitus (DM) type I and II as well as their close relatives to clarify correlations between circulatory disorders, disturbances of lipid metabolism (LM) and hemocoagulation. MATERIALS AND METHODS: The patients and their relatives were divided into 3 groups: with AS, DM type I and II. Individuals without relevant hereditary predisposition served control. A total of 564 subjects were examined. Thermography (TG), TV capillaroscopy of the nail bed (TVCNB), measurements of total blood cholesterol (TBC), high density lipoproteins (HDLP) cholesterol, triglycerides, coagulation time, plasma recalcification, free heparin, fibrinogen concentration, calculation of the atherogenic and prothrombin indices were conducted. RESULTS: Microcirculation suffered most of all, especially in patients and their relatives with AS and DM type I. TVCNB detected morphological changes of the capillaries confirmed by TG. DM patients and their relatives, especially those with DM type I had most distinct morphological shifts. CONCLUSION: The above PC disorders result from lipid metabolism and hemocoagulation abnormalities affecting blood rheology. In DM patients and their relatives especially in those with DM type I specific alterations of the capillary structure in DM and hereditary predisposition to DM are a contributing factor. PMID- 9864796 TI - [Arterial hypertension in diabetes mellitus: pathogenesis and treatment (review)]. PMID- 9864797 TI - [Role of autonomic nervous system in pathogenesis and clinical manifestation of arterial hypertension in Icenko-Cushing disease]. AB - AIM: To study the role of autonomic nervous system (ANS) in pathogenesis and clinical symptoms of arterial hypertension (AH) in Icenko-Cushing disease (ICD). MATERIALS AND METHODS: Clinical and paraclinical parameters (vegetative reflexes, special tables, 10 words memory and Spilberg's tests, ECG, electroencephalography, computed tomography, MR-tomography, hormonal status) were studied in 48 ICD patients before and after treatment (adenomectomy, irradiation of the hypophysis, adrenalectomy, parlodel, chlotidin chemotherapy). RESULTS: 94.5% of patients with active ICD had blood hypertension and vegetovascular asthenia. There was a trend to cerebrovascular impairment. In spite of biochemical and clinical ICD remission lipothemia persisted in many patients. CONCLUSION: Pathogenesis of blood hypertension in ICD is related to hypersecretion of ACTH and corticosteroids but further progress of the disease depends on phenotypic features of autonomic nervous system and developing defects of CNS. PMID- 9864798 TI - [Prevalence of carbohydrate metabolism disorders in free-living females of capable age in the city of Baku]. AB - AIM: The study of prevalence of carbohydrate metabolism disorders (CHMD) in females. MATERIALS AND METHODS: A representative sample of 20-59-year-old female population of the city of Baku was examined using age-specific standardized index, WHO tests and criteria. RESULTS: CHMD occurred rather frequently. Tolerance to glucose was abnormal in 50% of the examinees. Diabetes mellitus affected 20-29-year-old women 4 times more frequently than poor glucose tolerance. An opposite trend was observed for 40-49-year-old women. CONCLUSION: Relevant prophylactic measures are required for female Baku population. PMID- 9864799 TI - [Epidemiology of iodine deficiency in the middle Ob regions]. AB - AIM: Epidemiological rvaluation of goiter endemia and iodine deficiency in population of the Middle Ob region. MATERIALS AND METHODS: Thyroid palpation and ultrasonic investigations, iodine measurements in the urine were made in 6256 children and adolescents aged 8-17 years. RESULTS: Euthyroid condition was clinically stated in all the examinees. Palpation discovered thyroid enlargement in 36.78% of the examinees. Age prevalence was: 27.3, 33.28, 42.8, 46.18% in children and adolescents aged 8-10, 11-12, 13-14 and 15-17 years, respectively. Ultrasound detected goiter in 30.6% of the examinees. The occurrence was 34.78, 31.68, 28.2, 19.8% in the above age groups, respectively. No significant differences by thyroid size existed in girls versus boys. Urinary iodine was low in all the age groups. CONCLUSION: The Middle Ob regions belong to zone of goiter endemia with mild to moderate iodine deficiency. This dictates the necessity of introduction of special programs of iodine deficiency control and prevention. PMID- 9864800 TI - [Gestational arterial hypertension: clinico-epidemiological study]. AB - AIM: Epidemiological assessment of prevalence of blood hypertension (BH) in pregnant women. MATERIALS AND METHODS: Blood pressure and urinary protein were measured in 5160 gravidae observed in female consultations in 4 cities of Russia. RESULTS: BH was found in 1042(20.2%) examinees. Chronic BH was diagnosed in 306 patients. The rest females had gestational BH. Essential hypertension occurred in 174 patients. CONCLUSION: A stable rise of diastolic blood pressure above 90 mm Hg is a reliable diagnostic criterion of hypertension in gravidae. Classic proteinuric forms of gestational BH are rare. PMID- 9864801 TI - [Replacement hormonal therapy: effects on ECG parameters in postmenopausal women]. AB - AIM: To examine postmenopausal women free of organic cardiovascular lesions receiving replacement hormonotherapy (RHT) for ECG abnormalities, control of the detected cardiovascular defects. MATERIALS AND METHODS: ECG was conducted in 61 postmenopausal women before and during RHT with climen and climonorm. RESULTS: ECG registers frequent functional abnormalities in postmenopausal women. A 3 month RHT improved the patients' condition. ECG parameters improved on RHT month 9. CONCLUSION: RHT of postmenopausal women corrects general condition of such patients as well as functional ECG parameters. PMID- 9864802 TI - [Benign breast diseases: diagnosis and treatment (lecture)]. PMID- 9864803 TI - [Present-day diagnostic techniques in breast nodes]. AB - AIM: A comparative analysis of the effectiveness of preoperative diagnostic techniques in breast nodes. MATERIALS AND METHODS: 110 females with breast cancer and 306 females with benign tumors of the breast were examined (mammography, ultrasound, aspiration biopsy). A new technique of ultrasound histography of the tumor which enhances diagnostic specificity of echography is introduced. RESULTS: Diagnostic sensitivity averaged 82.8 +/- 3.9, 88.3 +/- 3.2, 69.3 +/- 4.5% for mammography, ultrasonic visualization and aspiration biopsy, respectively. Informative value of mammography and aspiration biopsy decreased in females of reproductive age, especially in preexisting hyperplastic processes in the breast. CONCLUSION: With greater size of the tumor in the breast ultrasound becomes more sensitive but less specific. The additional histography of the tumor in such cases is useful. PMID- 9864804 TI - [Coagulation system in patients with blood malignancies treated with estrogens in the course of cytostatic therapy]. AB - AIM: To evaluate the effect of estrogen gestagenic drugs (EGD) on blood coagulation in patients with blood malignancies treated by polychemotherapy. MATERIALS AND METHODS: 32 females of reproductive age treated with ovarioprotector marvelon during cytostatic therapy. The following indices were determined: activated partial thrombin time, prothrombin index of the ethanol test, fibrinogen and antithrombin III levels, factor XIIa dependent time of fibrinolysis, thrombin time. Dopplerography of the lower limb vessels was carried out. RESULTS: No significant alterations in coagulograms of the patients treated with cytostatics in combination with EGD were recorded. CONCLUSION: It is safe to use EGD as ovarioprotectors in females with hematological malignancies in hypercoagulation. PMID- 9864805 TI - [Heptral potentialities in the treatment of hepatic damage in children with malignant blood disease]. AB - AIM: To assess benefit for children with malignant blood disease (MBD) of the hepatoprotector heptral. MATERIALS AND METHODS: 67 children with blood malignancy aged 3-14 years were examined (53 of them had acute lymphoblastic leukemia). 39 patients were in the study group (25 of them had hepatitis B or C), 28 were controls. Initially, heptral was injected intravenously (14 days) then orally (16 30 days). Activity of transaminases and number of violations of polychemotherapy protocols because of hepatic toxicity were registered. RESULTS: Heptral administration led to inhibited activity of AlAT and AsAT especially in non infected patients. Protocol deviations became less frequent. CONCLUSION: Heptral is a potent hepatoprotector in hepatic lesions of toxic, viral and mixed origin. PMID- 9864807 TI - [Alcohol abuse in Russia is an essential risk factor of cardiovascular diseases development and high population mortality (review)]. PMID- 9864806 TI - [Clinical trial of domestic drug polyoxidonium in secondary immunodeficiency]. AB - AIM: To test clinical efficiency and immunocorrectives characteristics of polyoxidonium in patients with chronic bronchitis (CB) and chronic recurrent furunculosis. MATERIALS AND METHODS: 56 patients entered the trial; 13 with CB in remission, 22 with CB in exacerbation and 21 with chronic recurrent furunculosis. Clinical examinations and tests were made before polyoxidonium therapy and 2-3 days after it. Immunological tests consisted of determination of sub-population composition of peripheral blood lymphocytes, serum immunoglobulins and phagocytic activity of neutrophils. RESULTS: Polyoxidonium is an effective and safe immunocorrector. It has a strong influence on functional activity of blood phagocytic cells. CONCLUSION: Polyoxidonium treatment of CB and chronic recurrent furunculosis results in positive shifts in hematological and immunological indices. This allows to recommend it for treatment of acute and chronic infections. PMID- 9864809 TI - [Pneumomycoses (lecture)]. PMID- 9864808 TI - [Clinical manifestation, diagnosis and treatment of rare forms of acute drug poisoning (lecture)]. PMID- 9864810 TI - [Clinical pharmacology of antidepressants (review)]. PMID- 9864811 TI - [Autonomic effect of grandaxin in the treatment of neurotic disorders in general somatic practice (lecture)]. PMID- 9864812 TI - [Isoptin. Evolution of views from Fleckenstein to the present days]. PMID- 9864813 TI - [Experience in heptral treatment of diffuse liver diseases]. AB - AIM: Heptral trial in alcoholic, drug and viral diseases of the liver. MATERIALS AND METHODS: 67 patients with chronic diffuse liver diseases were treated with heptral. The examination covered AlAt, AP, HHTP, bilirubin, cholesterol, CT of the liver, esophagogastroduodenoscopy. Heptral was given by two steps: 14 days of intravenous drops (800 mg/day) followed by 14 days of oral use (2 tablets, 400 mg each) before meal at 8 a.m. and 2 p.m. RESULTS: In alcoholic disease of the liver, heptral relieved depression, reduced AlAT, AP, HHTP, bilirubin, density of the liver. The addition of heptral to interferon-alpha-2 treatment of chronic hepatitis C corrected intrahepatic cholestasis. In drug-induced liver damage heptral promoted normalization of the hepatic tests, improved general condition of the patients. CONCLUSION: Heptral is effective in patients with alcoholic and drug liver lesions, chronic hepatitis C. PMID- 9864814 TI - [Enzyme preparations in conservative treatment of chronic pancreatitis]. AB - AIM: To study effectiveness of enzyme medicine pancitrate 25,000 in the treatment of chronic pancreatitis (CP). MATERIALS AND METHODS: Pain, stool, creato- and steatorea, other signs of CP were examined before and after pancitrate treatment of 17 patients with chronic pancreatitis aged from 28 to 62 years. RESULTS: Pancitrate 25,000 reduced stomach pains in 16 of 17 patients. Stool returned to normal in 8 of 13 patients. Creato- and steatorea decreased. CONCLUSION: New dosage forms of the enzyme preparations (pancitrate 25,000, in particular) are highly effective both in deficiency of pancreatic external secretion and pain relief in exacerbation of CP. PMID- 9864815 TI - [Effect of nocodazole on endocytosis of epidermal growth factor receptor]. AB - During endocytosis EGF-receptor complexes are transported from early peripheral endosomes to late juxtranuclear-located endosomes to be then degraded in lysosomes. It is suggested that such a spatial organization of endosomal compartments is maintained by microtubule system and is necessary for lysosomal degradation of endocytosed cargo. In the present work, a study was made of the influence of Nocodazole, a microtubule depolymerizing agent, on endocytosis of fluid phase marker HRP and EGF entering the cell via receptor-mediated endocytosis. By subcellular fractionation in Percoll gradient it was shown that Nocodazole did not affect HRP internalization but stimulated its accumulation in a fraction sedimented together with late endosomes, thus preventing HRP delivery to lysosomes. On the contrary, Nocodazole exerted no influence on dynamics of compartmentalization and lysosomal degradation of EGF-receptor complexes. Moreover, no alterations were found in the functioning of a so well-known EGF stimulated signal transduction pathway as MAP-kinase cascade. At the same time microtubule depolymerization dramatically changed the morphology of endosomal compartments abolishing juxtranuclear localization of late endosomes. Our data suggest that translocation of EGF-receptor complexes is not necessary for their normal lysosomal processing. Rab7, traditionally considered as a marker of late endosomes, has been found to demonstrate in Nocodazole-treated cells, in contrast to the control, a low extent of co-localization with endosomal structures. It could be supposed that the role of Rab7 is not so much to mediate early-to-late endosome transition as to maintain spatial organization of endosomal apparatus by mediating endosome-cytoskeleton interactions. PMID- 9864816 TI - [Studies of small GTPase Rab7 association with endosomes of cells expressing normal and mutant forms of epidermal growth factor receptors]. AB - A study was made of an association of small GTPase Rab7, commonly considered as a marker of late endosomes, with endosomal compartments of cells expressing EGF receptor with different ability to be sorted for degradative pathway. It was found that in cells HER14, expressing normal EGF receptor, Rab7 was associated with both early and late endosomes and the extent of association correlated with the number of EGF-receptor complexes in the specific endosomal fraction. Cels with a receptor, lacking major sites of autophosphorylation by deletion of 126 C terminal residues (CD126), demonstrated a low efficiency of EGF-receptor sorting to late endosomes and decreased association dynamics with Rab7. Interaction of Rab7 with endosomes of cells expressing kinase negative receptors (K721) was found to be minimal. At the same time, in cells Cd126 and K721 with a low sorting efficiency Rab7 was mainly associated with early endosomes. These data favor Rab7 involvement in mediating early-to-late endosomal transition. PMID- 9864817 TI - [Dynamics of various integral indices of individual changes of heat-resistance of isolated frog muscles kept in Ringer solution]. AB - A study was made of the dynamics of the heat resistance (HR) mean level, of the correlation between the initial individual HR level and the value and direction of this level shifting, and of the pattern of correlation between HR levels of pair sartoria muscles and variability of HR level in muscles kept at 5-6 degrees C in the Ringer for 12 days. A two-phase changes in the mean HR level of muscles was revealed that involved its increase by 48% within one day of maintenance in the Ringer solution followed by its decrease by 14% in 6 days. On the first steps of muscle maintenance in the Ringer solution, a negative correlation was noticed between the initial HR level and the value and direction of its shifting, in addition to narrowing the range of variability of muscle HR level. All these events were accompanied by phase fluctuations in the correlation coefficient between HR levels of pair muscles. By the end of experiment the range of variability of the HR level was seen to increase, and the earlier positive correlation between pair muscle HR levels appeared either to change or to be a negative one. The increase in the range of variability preceded the decrease in the mean level of muscle HR. PMID- 9864818 TI - [Contact inhibition of desmin expression in rat liver Ito cells in vitro]. AB - The presence of desmin is used to identify the Ito cells in rat liver and to evaluate the purity of separated and cultured cells. The present study was conducted to evaluate the expression of cytoskeletal (with special emphasis on desmin) and intercellular matrix proteins in cultured rat liver Ito cells. The Ito cells were isolated by enzymatic digestion of rat liver and purified by single-step Nycodenz gradient. During the first week of culturing, the Ito cells proliferated and all of them expressed desmin and vimentin. By the end of the first week, part of them were seen to transdifferentiate into myofibroblasts. These cells expressed alpha-smooth muscle actin. The Ito cells and myofibroblasts were stained with anti-laminin, and anti-collagen I and III antibodies. The expression of desmin in myofibroblasts was less than in untransformed (stellate shaped) Ito cells. By the end of the second week the cells developed a dense monolayer and lost desmin. We suggest that repression of desmin in the cablestone like monolayer of cells may be due to cell-to-cell contact inhibition. PMID- 9864819 TI - [Chromatin compactification using a model system of DNA-protein complexes]. AB - Conformational peculiarities of DNA complexes with histones of the H1 family have been studied by the method of circular dichroism (CD). The H1 histones were isolated from spermatozoa of the sea urchin Strongylocentrotus intermedius, starfish Aphelasterias japonica, and bivalve mollusc Chlamis islandicus and also from the rat thymus. It is shown that these sperm-specific histones do not compact DNA in low ionic strength solution. At physiologic conditions H1 from the sea urchin and starfish sperms compact DNA more intensively than other histones. The H1 from rat thymus has a minimum ability to compact DNA. This histone does not change the structure of DNA double helix. It was supposed that it could be associated with interactions of this histone with DNA in the major groove of its helix. At the same time sperm-specific H1 can interact with DNA not only in the major groove but also in the minor groove, and this induces changes in DNA structure. This DNA-protein interaction is specific for the sperm chromatin and may support the supercompact organization of the sperm chromatin. PMID- 9864820 TI - [Electron microscopic in situ hybridization of dUTP-digoxigenin labeled probes with polytene chromosomes of Drosophila melanogaster]. AB - Using DNA probes labeled with digoxygenin-11-dUTP, a simplified method of electron microscopic (EM) in situ hybridization was developed for standard squashes of Drosophila melanogaster polytene chromosomes. The developed method is efficient and reproducible: its high resolution and specificity was shown for the transformed strain 148, in which the insertion was localized by EM as a new thin band. The method was applied for fine mapping of the developmentally regulated complex gene, muscleblind (mbl), which was shown to cover the 54B1-2 large band and the adjacent interbands in 2R polytene chromosome. PMID- 9864821 TI - [Process of active dissociation of vital stain Hoechst 33342 from DNa in cells of cultured rodent cell lines]. AB - The process of active dissociation or "DNA clearing" of not covalently binding agents from DNA in living HeLa cells was shown by the flow cytometry technique. The vital fluorescent bisbenzimidazole dye Hoechst 33342, which binds tightly but not covalently to DNA in a minor groove, was used as a basic model to study the interaction of not covalently binding agents with DNA. In this paper, we continue to analyse the "DNA clearing" process in the living fibroblasts of different rodent species (mouse, rat, Chinese hamster). The obtained data suggest that the processes of active dissociation or "DNA clearing" of Hoechst 33342 have some common features in all investigated mammalian cells. Nevertheless, some differences in the process were found in these lines. The role of nucleotide excision repair genes in the process of DNA clearing was not established. PMID- 9864822 TI - [Potential of the image analyzer "videotest" for performing microphotometric study in cytology]. AB - Precision features of image analyser "Videotest" and possibilities of its applications for cytophotometric investigations have been analysed. It has been shown that a photometric feature is linear at least to optical density 1.1. Cytophotometry of the Feulgen stained rat hepatocyte nuclei has been performed. Coefficients of variation of DNA for 2c, 4c and 8c nuclei are 7.72, 7.65 and 6.50%, respectively, and ratios between average values of these nuclei are 1.00:2.06:4.12. Possible causes of errors of digital television equipment for cytophotometry are discussed. PMID- 9864823 TI - [The current epizootic situation of prospects for eradicating rabies]. PMID- 9864824 TI - [Comparative characteristics of some carnivore parvoviruses]. PMID- 9864825 TI - [Enteroviral persistence in eye infections in monkeys (postuveitis syndrome)]. AB - Follow-up (5.5 years) of monkeys with a history of acute enterovirus uveitis (AEU) after intraocular infection with ophthalmotropic ECHO 11 and ECHO 19 strains showed progressive changes in the infected eye; destruction of the iris, formation of films in the anterior chamber of the eye, deformation of the pupil, and signs of cataract and glaucoma. A short (1-30 days) stage of active reproduction of infective virus (up to 10(10) TCD50/g tissue in the first days postinfection) was followed by the second very long stage of limited specific multiplication of virus in ocular tissues. The virus was not detected during the second stage, but its components (virusspecific antigen and crystalloid accumulations of virions) were found in ophthalmic and conjunctival tissues; moreover, high levels of neutralizing antibodies were detected in the blood. The results indicate AEU transformed into a stubborn persistent infection of the eye in monkeys. The findings of this follow-up and published reports about many-year follow-up of children with AEU permit us to classify a post-uveitis disease as a special nosological form of a persistent enterovirus infection and denote it as enterovirus post-uveitis syndrome. The main signs of this syndrome are a history of AEU, progressive destructive changes in the anterior segment of the eye, presence of virus components in ocular tissues in remote periods after infection, and high antibody level in the blood. PMID- 9864826 TI - [Antigenic activity of delta-antigen peptides corresponding to three genotypes of the delta hepatitis virus]. AB - Six synthetic peptides from immunodominant region 65-80 aa derived from the sequences of delta-antigen corresponding to 3 HDV genotypes were obtained. Their antigenic activities were assessed by indirect enzyme immunoassay with sera of patients with hepatitis D and of asymptomatic carriers of anti-delta antibodies. Differences in antigenic characteristics of full-size 65-80 and truncated 71-80 aa peptides corresponding to three HDV genotypes were revealed. The diagnostic significance of synthetic 65-80 aa fragments is shown. PMID- 9864827 TI - [Serological properties of HIV-1 isolates from the focus of an epidemic in the Gomel region of the Belarus Republic (1996)]. AB - Immunoreactivity of sera obtained in 1996 from HIV-1-infected intravenous narcomaniacs from an epidemic focus in Gomel region, Belarus, is studied with a panel of 10-16-component synthetic peptides simulating apical epitope of surface glycoprotein gp120 V3 loop of HIV-1 variants. Comparative analysis of resultant spectra with representative immunoreactivity spectra of sera from a sampling of HIV-1-positive sera collected in 1986-1997 in different regions of the former USSR demonstrated a high homogeneity of immunoreactivity spectra of sera from Gomel region. Serotypes of HIV-1 A/C variants circulating in populations of intravenous narcomaniacs in the focus in Gomel region in 1996 and in Southern and South-Eastern Ukraine in 1995-1996 are similar. This confirms a previous conclusion about the prevalence of A/C serotype among intravenous narcomaniacs in the former USSR. PMID- 9864829 TI - [Analysis of core virion polypeptides from the pathogen causing chicken egg-drop syndrome]. AB - The cores of egg-drop syndrome virus (EDS-76) were isolated by the pyridine technique. EDS-76 proved to be much more resistant to pyridine disruption than other adenoviruses and treatment with 10% pyridine did not lead to complete dissociation of capsid and cores; only increase of pyridine concentration to 20% produced satisfactory results. At least three polypeptides (24, 10.5, and 6.5 kDa) were found in the core by SDS-PAGE, whereas the 40 kDa reacting with the core is most probably not a core component. Much more intensive reactions of the core with EDS-76 virion capsid suggest that its virion structure differs from that of other adenoviruses. PMID- 9864828 TI - [Indication and identification of California serogroup viruses in experimentally infected Ae. aegypti mosquitos by a lanthanum immunofluorescent analysis method]. AB - The sensitivity and specificity of lanthanide fluoroimmunoassay (LFIA) and enzyme immunoassay (EIA) for indication and identification of California antigens in suspensions of infected Aedes aegypti mosquitoes are compared. The sensitivity of LFIA was 2.3 Ig TCD50/ml, EIA was less sensitive and detected virus antigens in mosquito suspensions at infective titer of 4.1 Ig TCD50/ml. If a pool contained mosquitoes infected with different California viruses, they could be identified only at the level of serological complex. PMID- 9864830 TI - [Comparative analysis of tests on in vitro and in vivo quantitative assessment of the immunogenicity of tick-borne encephalitis vaccine]. AB - Many-year routine use of EIA as an in vitro test demonstrated it as a highly reproducible and technological test for assessing the efficacy of vaccine against tick-borne encephalitis and its semiproducts at the intermediate stages of vaccine production. The reproducibility of mouse protection test is notably inferior to that of EIA. PMID- 9864831 TI - [Little known pages from the history of discovering tick-borne encephalitis]. PMID- 9864832 TI - [Retroperitoneal fibrosis and arthritis--a manifestation of the same illness]. AB - Retroperitoneal fibrosis is a disorder in which the retroperitoneal fat is the site of a subacute and chronic inflammatory reaction and is subsequently replaced by dense fibrotic tissue. Rheumatoid nodules are chronic granulomata occurring at sites of pressure and movement, both near the body surface and internally. A 55 year-old sales-manager was admitted to radiation synovectomy after a 5 year history of excessive right and left knee effusions. There were no other clinical or laboratory abnormalities. The patient did not respond to either radioisotope synoviorthesis using radioactive Yttrium (90 Y), or to open synovectomy and prostetic surgery of the right knee. One year later, surgery of left ureter was necessary. Histological findings revealed the diagnosis of Ormond's disease. Comparative histological studies of synovial membrane of knee and retroperitoneal tissues showed local necrosis, fibrin deposition, lining cell proliferation, and infiltration by lymphocytes. Diagnosis of arthritis complicated by retroperitoneal rheumatoid nodules and retroperitoneal fibrosis was made. Serum rheumatoid factor has been negative. For the last 3 years, the patient has been on successful therapy with azathioprine. Rheumatoid nodules of the retroperitoneum have vanished completely and frequency of knee effusions decreased. PMID- 9864834 TI - [Experimental therapy of rheumatoid arthritis with antibodies to tumor necrosis factor]. PMID- 9864833 TI - [Immunopharmacologic profile and therapeutic prospects of anti-TNF-alpha therapy]. AB - Clinical trials with monoclonal antibodies directed against TNF alpha (anti-TNF mAbs) and soluble TNF receptor fusion proteins (sTNFR-IgGs) have demonstrated that systemic and synovial trapping of TNF alpha results in long lasting anti inflammatory and anti-nociceptive effects in patients with rheumatoid arthritis. Clinical indices of inflammatory synovitis and laboratory parameters (CRP and ESR) respond to single and repeated administrations of anit-TNF alpha therapies in a dose-dependent fashion. Studies on the immuno-pharmacological profile in patients suggest evidence that TNF alpha trapping down-regulates the effector mechanisms involved in the immuno-inflammatory response in rheumatoid arthritis. Inhibition of PLA 2- and COX-2-derived pathways of mediators of inflammation (prostanoids and leukotrienes) decreases signs and symptoms of inflammatory synovitis such as joint swelling, tenderness and pain. Down-regulating of the cytokine-inducible adhesion molecules ICAM-1, VCAM-1 and ELAM-1 in endothelial cells and synoviocytes results in a marked inhibition of transendothelial migration of inflammatory and immune cells. A decrease of cytokine-regulated metalloproteinase expression results in normalization of circulating MMP-1 and MMP-3 levels. The effect of TNF alpha neutralization on mechanisms of rheumatoid joint destruction has the long-term potential for preventing or decreasing the rate of erosive changes of cartilage and bone. PMID- 9864835 TI - [Therapy of refractory chronic polyarthritis with tumor necrosis factor alpha receptor fusion proteins (TNFR55-IgG1)--results of double-blind placebo controlled studies over 3 months]. AB - AIM OF STUDIES: To determine the optimal dose regimen for i.v. TNFR55-IgG1 in refractory rheumatoid arthritis. METHODS: 218 patients with refractory rheumatoid arthritis were enrolled for two double-blind placebo-controlled multicenter trials (Europe and USA). They were treated with monthly i.v. placebo, 0.01, 0.05, 0.1 or 0.5 mg/kg TNFR55-IgG1 after a 4 week wash-out of DMARDs. An additional German trial compared biweekly i.v. 20 mg TNFR55-IgG1 and monthly 50 mg following a loading does of 100 mg in 60 patients. RESULTS: TNFR55-IgG1 induced a substantial improvement already apparent one day after the first infusion. A maximal, dose dependent effect was reached after two weeks. Later, efficacy declined in parallel to an increase in anti-TNFR55-IgG1 antibodies resulting in an increased drug clearance. The drug was well tolerated with predominantly mild or moderate adverse events. CONCLUSION: Intravenous TNFR55-IgG1 was well tolerated and effective in refractory rheumatoid arthritis but the treatment schedules tested could not stabilise the initial improvement. PMID- 9864836 TI - [Therapy of refractory chronic polyarthritis with tumor necrosis factor alpha receptor fusion protein (TNFR55-IgG1)--long term follow-up of 80 personal cases]. AB - AIM OF THE STUDY: To investigate the long-term effect of TNFR55-IgG1 in patients with severe rheumatoid arthritis refractory to multiple previous DMARD treatments. METHODS: Between 1994 and 1995 we recruited 80 patients for two double blind, placebo-controlled multicenter trials. All patients were treated for 2-30 months with a glycosylated fused protein consisting of two human p55 TNF receptors linked to a human IgG1-Fc. The dose range was 2.5-100 mg/month given i.v. with cumulative doses from 40-940 mg. All patients were followed-up prospectively for 24-36 months after initiation of therapy. The evaluations included a 48 swollen and tender joint count, ESR. CRP, RF, ANA, ENA, and safety parameters at 6 month intervals. In addition, pharmacokinetics, TNF, and anti TNFR55-IgG1 antibody levels were available after the first and third injection. RESULTS: Data from 80 patients are available representing an experience of more than 170 patient years; 11 patients are still being treated. Predominant reasons for treatment withdrawal were restrictions in treatment protocol. Six patients died (1 during treatment, 5 in the post-treatment follow-up). All deaths were related to preexisting cardiovascular disease except one post-surgery septic arthritis 6 months after the last dosing (during vacation in another country). No malignancies were detected. Anti-TNFR55-IgG1 antibodies correlated with a decrease in drug half-time. Shifts in the TNFR55-IgG1 glycosylation pattern affected pharmacokinetics and efficacy. A lupus nephritis occurred 18 months after the last dosing in one patient with erosive RA (ANA and dsDNS positive already before TNFR55-IgG1 initiation). Continued treatment for 3 years in 11 patients resulted in an 81% reduction in swollen joint count, less morning stiffness, less pain, and a reduction in steroids. CONCLUSION: Treatment with TNFR55-IgG1 is safe and efficacious over the long-term in patients with severe refractory RA. PMID- 9864837 TI - [Treatment of chronic polyarthritis with a human recombinant interleukin 1 receptor antagonist]. PMID- 9864838 TI - [Anti-CD4 therapy in treatment of rheumatoid arthritis--have the die been cast?]. AB - The hypothesis of rheumatoid arthritis (RA) as a T cell mediated disease has led to the development of numerous therapeutic approaches that target the function of T cells. The development of monoclonal antibodies against the T cell surface molecule CD4 has raised hopes to achieve a major progress in the treatment of RA. However, after encouraging results in early open studies, double blind trials were unable to demonstrate the efficacy of anti-CD4-therapy in RA. There are numerous reasons to explain the failure of this treatment approach. Besides the fact that the T cell hypothesis of RA has repeatedly been challenged, pharmacological problems or an inappropriate selection of outcome criteria have to be considered. The final evaluation of anti-CD4 therapy in RA will be possible only after the testing of newly developed non-depleting anti-CD4 antibodies. PMID- 9864840 TI - [Health policy aspects of quality assurance]. AB - Health policy in Germany has in recent years given priority to the self administration of physicians, sickness funds, and hospital associations. Hence, legal regulations on quality assurance have focused on defining responsibilities and areas of primary importance, while precise processes are being defined by the self-administration. As prices for health care are increasingly restricted by lump sum payments, quality and quality assurance become more important as a factor in competition between health care providers. To support the implementation of quality assurance in the health care system, the German Federal Ministry of Health has funded model-projects, e.g., to support the adaptation of successful strategies of total quality management for health care. PMID- 9864841 TI - [Legal and occupational law principles of quality assurance]. PMID- 9864842 TI - [Quality management in medicine--definitions and goals]. AB - The meanings of the terms "quality", "quality of care", and "quality management" were clarified with common international definitions. Based on that, the basic ideas, concepts, and objectives of quality management in medicine were explained. Finally some future trends related to quality were viewed, which represent challenges for the medical profession and entire medicine. PMID- 9864843 TI - [Cost-effectiveness--limits between optimization and rationing]. AB - Although rationing is an important feature in the German health care system, this topic is not discussed in an explicit and structured way. Currently, most rationing decisions are not based on systematic data on costs and outcomes of diseases and medical interventions. A major task of future research will be to provide health policy makers with data on direct costs, indirect costs, effectiveness, and utility of medical interventions and to integrate these data into decision analysis models. Interdisciplinary work-groups should be set up, combining clinical and economic knowledge. On the other hand, official grants are needed to perform unbiased research in this field covering the various areas of medical care. PMID- 9864844 TI - [Quality management in medical rehabilitation]. AB - In the context of the quality assurance program of the German pension insurance institutions, instruments and procedures have been developed in the past few years in order to assess the quality of structure, process, and results of inpatient rehabilitation, as well as patient satisfaction. The routine implementation of these procedures aims at providing rehabilitation hospitals with fast information on shortcomings in their current practice, so that concepts of treatment and--in the long run--effects of rehabilitation can be systematically improved. When the "quality profiles" of rehabilitation hospitals are compared ("benchmarking"), paying institutions of rehabilitation could gain information on the cost-quality-ratio of particular hospitals as well as important leads for improving the steering processes of patients and hospitals. In the present article, results of a first "field-run" of the procedures for measuring patient satisfaction and quality of rehabilitation processes are reported. A comparative analysis of patient satisfaction in four orthopaedic/rheumatologic rehabilitation hospitals showed high degrees of overall satisfaction in all hospitals. In several subdimensions of patient satisfaction, however, significant differences between hospitals were found. Quality of rehabilitation processes--as assessed by a standardized peer-review--varied substantially between 36 hospitals. Problems of quality were found especially in some dimensions that must be considered as crucial for rehabilitation, e.g., in the history and diagnosis of disabilities, in the consideration of psycho-social problems, or in the assessment of occupational capacities. PMID- 9864845 TI - [BAK/KBV/AWMF curriculum "Quality assurance--medical quality management": concept of goal-oriented conversion to general practice applications]. AB - Since 1996, the curriculum "Quality management for physicians" exists with a three-level training concept. Based on these contents, a purposeful course program has been developed by the ZQ, and through May 1998 four level I-, four level II- and four level III-courses have been performed in Hannover. Thus, 50 physicians are qualified for new professional roles as hospital quality managers. The evaluation of a standardized questionnaire shows that the concept is highly accepted--meanwhile also by other professionals in health care. As a new model of quality assurance/quality evaluation is now developed for German hospitals, the contents of the training program will be updated for actual needs. PMID- 9864846 TI - [Quality recommendations of the German Society of Rheumatology. Basis for further responsibilities]. AB - The manual "Qualitatssicherung in der Rheumatologie" is a consensus based thesaurus of diagnostic and therapeutic guidelines. The Committee for Quality Assurance in the German Society of Rheumatology proposes a plan for further activities in this field. The revision and updating of the manual was already begun, which will be continued and shall be Internet-based in the near future. Guidelines with topics of interdisciplinary interest (e.g. low back pain, ankylosing spondylitis) are to be unified. In order to achieve a broader consensus, practicing physicians and self-help organizations of patients should be asked to review the guidelines. Some controversies in clinical rheumatology are to be solved in prospective studies. Whereas the manual is kept as a body of reference for rheumatologists, a practice guideline "joint swelling" will be developed according to recommended criteria for appraisal of guidelines. PMID- 9864847 TI - Invention and mystery in hypotrich DNA. AB - Hypotrichs have evolved extraordinary ways of organizing, manipulating, and replicating the DNA in their micronuclear and macronuclear genomes. Short macronuclear DNA molecules containing single genes are created by excision from chromosomes, accompanied by massive elimination of the germline DNA sequences between genes. Germline genes themselves are interrupted by multiple noncoding segments called internal eliminated segments, or IESs, that divide genes into multiple macronuclear-destined segments, or MDSs. The functional significance of this organization is unknown. Over evolutionary time IESs accumulate mutations rapidly, are inserted into or excised from genes, and shift position along DNA molecules. MDSs are ligated to create functional genes when IESs are spliced out of micronuclear DNA during macronuclear development. MDSs in some germline genes are in scrambled disorder and become unscrambled in association with IES elimination. Replication of DNA in the macronucleus is accomplished by organization of replication enzymes and factors into a structure that sweeps through the macronucleus to replicate the many millions of gene-sized DNA molecules. The significance of many of the bizarre DNA phenomena in the evolutionary/functional success of hypotrichs is still unclear. PMID- 9864848 TI - Introducing antisense oligodeoxynucleotides into Paramecium via electroporation. AB - A method utilizing electroporation to deliver antisense oligodeoxynucleotides into Paramecium tetraurelia has been developed. For these studies antisense oligonucleotides directed to different regions of the calmodulin mRNA were used. It was found that a pulse delivered at 150-250 V (375-625 V/cm field strength) for 3.9-4.2 ms using a 275 microF capacitor with resistance set at 13 Ohms was sufficient to achieve measurable incorporation of fluorescently-labeled oligodeoxynucleotides in up to 95% of the cells treated. Optimal parameters included using oligodeoxynucleotides of at least 12 bases in length with a 3' blocking group at a dose of around 10 microM. In addition, multiple oligodeoxynucleotides directed to the same target mRNA resulted in at least a 10 fold reduction in the dose of oligodeoxynucleotide required to achieve the desired effects. Taken together, these results indicate that the use of antisense oligodeoxynucleotides can be an easy and useful method for linking genes to specific functions in Paramecium tetraurelia. Finally, this report discusses how different 3' blocking groups and the use of combinations of oligodeoxynucleotides directed to different regions of the same target mRNA can help address concerns about specificity. PMID- 9864849 TI - The HSP70 gene family in Pneumocystis carinii: molecular and phylogenetic characterization of cytoplasmic members. AB - Pneumocystis carinii, a major opportunistic lung pathogen of AIDS patients, is found in a number of mammals and is proposed to be a member of the fungi. In this work, several members of the highly conserved HSP70 multigene family were characterized from rat-derived P. carinii. Previously, we reported characterization of the ER resident HSP70 homolog known as BiP from prototype (P.c. carinii) and variant (P. c. rattus) strains of the organism. We report here, from P. c. carinii, characterization of Pcsa1, an HSP70 homolog that encodes a cognate/stress-induced HSP70 homolog of the SSA subfamily in Saccharomyces cerevisiae. We also identify, from both rat strains and from a human isolate of P. carinii (P.c. hominis), a third set of HSP70 homologs that apparently encode a ribosome-associated cytoplasmic HSP70 homologous to the S. cerevisiae SSB subfamily. Our data indicate that Pcsal mRNA, like Pcbip mRNA, bears an intron in the 5' untranslated region, is induced by heat shock, and suggest that this gene undergoes alternative transcription and splicing. The SSB homologs display significant sequence heterogeneity between P. carinii source strains, supporting the genetic divergence and likely speciation of P. carinii isolates within and between host species. Phylogenetic analysis with the PcSA1 protein supports inclusion of P. carinii among the higher fungi. PMID- 9864850 TI - Characterization of myosin heavy chain and its gene in Amoeba proteus. AB - Monoclonal antibodies against the myosin heavy chain of Amoeba proteus were obtained and used to localize myosin inside amoebae and to clone cDNAs encoding myosin. Myosin was found throughout the amoeba cytoplasm but was more concentrated in the ectoplasmic regions as determined by indirect immunofluorescence microscopy. In symbiont-bearing xD amoebae, myosin was also found on the symbiosome membranes, as checked by indirect immunofluorescence microscopy and by immunoelectron microscopy. The open reading frame of a cloned myosin cDNA contained 6,414 nucleotides, coding for a polypeptide of 2,138 amino acids. While the amino-acid sequence of the globular head region of amoeba's myosin had a high degree of similarity with that of myosins from various organisms, the tail region building a coiled-coil structure did not show a significant sequence similarity. There appeared to be at least three different isoforms of myosins in amoebae, with closely related amino acids in the globular head region. PMID- 9864851 TI - Efficacy of novel antimicrobials against clinical isolates of opportunistic amebas. AB - We examined the effects of the macrolide antimicrobial agent azithromycin and phenothiazine compounds against clinical isolates of Acanthamoeba spp. and Balamuthia mandrillaris, opportunistic pathogens of human beings and other animals. Acanthamoeba growth was inhibited in vitro at 1, 5, and 10 micrograms/ml of azithromycin, but not the macrolides, erythromycin, and clarithromycin. In experiments attempting to simulate in vivo conditions, azithromycin protected monolayers of rat glioma cells from destruction by Acanthamoeba at a concentration of 0.1 microgram/ml, and delayed destruction at concentrations of 0.001 and 0.01 microgram/ml. We concluded that the minimal inhibitory concentration of azithromycin was 0.1 microgram/ml. Our results, however, suggested that the drug was amebastatic but not amebicidal, since ameba growth eventually resumed after drug removal. The phenothiazines (chlorpromazine, chlorprothixene, and triflupromazine) inhibited Acanthamoeba growth by 70-90% at 5 and 10 micrograms/ml, but some of these compounds were toxic for rat glioma cells at 10 micrograms/ml. Azithromycin was not very effective against B. mandrillaris in an in vitro setting, but was amebastatic in tissue culture monolayers at concentrations of 0.1 microgram/ml and higher. Balamuthia amebas showed in vitro sensitivity to phenothiazines. Ameba growth was inhibited 30-45% at 5 micrograms/ml in vitro, but completely at 5 micrograms/ml in the rat glioma model. In spite of their potential as antiamebic drugs in Balamuthia infections, toxicity of phenothiazines limits their use in clinical settings. PMID- 9864852 TI - Infraciliary bands in the rumen ophryoscolecid ciliate Ostracodinium gracile (Dogiel, 1925), observed by light microscopy. AB - Morphological features of the rumen ciliate Ostracodinium gracile (Dogiel, 1925) are described from pyridinated silver carbonate-impregnated specimens. Ostracodinium gracile has a characteristic arrangement of infraciliary bands not present in other ophryoscolecid ciliates. Buccal infraciliature is composed of three polybrachykineties. The adoral polybrachykinety does not completely encircle the circumference of the vestibular opening, but arches ventrally from its right to left side. The dorsoadoral polybrachykinety extends laterally along the dorsal side of the vestibular opening. The vestibular polybrachykinety extends along the dorsal wall of the long tubular vestibulum. Dorsal infraciliature consists of the dorsal polybrachykinety that extends laterally along the dorsal side of the body. During binary fission, four primordia, that is ventral, right, left, and dorsal primordia, form in the stomatogenic field and develop into the adoral, dorsoadoral, vestibular, and dorsal polybrachykineties of the opisthe, respectively. PMID- 9864853 TI - Cloning of two putative Giardia lamblia glucosamine 6-phosphate isomerase genes only one of which is transcriptionally activated during encystment. AB - The biosynthesis of the carbohydrate component of the cyst wall of the protozoan parasite Giardia lamblia, a polymer of N-acetylgalactosamine (GalNac), is by a pathway that is initiated with the conversion of fructose 6-phosphate to glucosamine 6-phosphate by an aminating isomerase, glucose 6-phosphate isomerase. This enzyme appears only after Giardia trophozoites are induced to start the production of cyst wall components after bile is added. To investigate whether induction of glucosamine 6-phosphate isomerase is by protein modification or by transcription activation, its gene was cloned and sequenced. Two genes, gpi1 and gpi2, encoding putative glucosamine 6-phosphate isomerases were identified but one, gpi1 was expressed. The transcript for gpi1 appeared not earlier than 6 h after cells were induced with bile salts. These results show that the first enzyme in the pathway leading to GalNac synthesis in encysting Giardia cyst wall biosynthesis is under transcriptional control. PMID- 9864854 TI - Phylogenetic diversity of parabasalian symbionts from termites, including the phylogenetic position of Pseudotrypanosoma and Trichonympha. AB - The phylogenetic diversity of parabasalian flagellates from termite hindguts has been examined by small subunit ribosomal RNA (rRNA) amplification and sequencing. Two species of particular interest, the giant trichomonad Pseudotrypanosoma giganteum and the hypermastigote Trichonympha magna, were isolated from the gut of Porotermes adamsoni by micropipetting, and the rRNA genes from these small populations amplified and sequenced. rRNA genes representing Hypermastigida and the Trichomonadida families Devescovinidae and Trichomonadidae, were also recovered by amplification from whole hindguts of three termites, P. adamsoni, Cryptotermes brevis, and Cryptotermes dudleyi. The parabasalian rRNA genes from C. brevis were found to comprise a unique and extremely heterogeneous lineage with no clear affinities to any known parabasalian rRNAs. In addition, one of the sequences isolated from P. adamsoni was found to be similar to another uncharacterised rRNA gene from Reticulitermes flavipes. The phylogeny of all known parabasalian small subunit rRNAs was examined with these new sequences. We find many taxonomic groups to be supported by rRNA, but not all. We have found the root of parabasalia to be very difficult to discern accurately, but have nevertheless identified several possible positions. PMID- 9864855 TI - Encephalitozoon hellem in two eclectus parrots (Eclectus roratus): identification from archival tissues. AB - Members of the phylum Microspora are obligate, intracellular, single-celled parasites identified in a wide range of vertebrate and invertebrate hosts. Only a few cases of microsporidial infections have been documented in psittacine birds including peach-faced, masked, and Fischer's lovebirds (Agapornis roseicollis, A. personata, and A. fischeri, respectively), budgerigars (Melopsittacus undulatus), and a double yellow-headed Amazon parrot (Amazona ochrocephala). Parasite identification has typically been limited to phylum or genus, and no avian species of microsporidia has clearly been described. In this report, microsporidia were identified in the kidney and intestine of a new host, the eclectus parrot (Eclectus roratus). Parasites were identified as Encephalitozoon hellem using morphologic, ultrastructural, and small subunit ribosomal RNA gene sequence data obtained from archived tissues. This parasite species was first identified in immunocompromised humans and may be a potential zoonotic pathogen. The epidemiology and prevalence of this parasite in humans and birds should be further explored. PMID- 9864856 TI - Ribosomal DNA fragments enhance the stability of transfected DNA in Entamoeba histolytica. AB - Genetic manipulation of Entamoeba histolytica is limited by the inability to express foreign genes at high levels. We tested whether sequences from the E. histolytica rDNA episome, present in 200 copies per cell, could act to stabilize the episomal transfection vector pTCV1. Ligation of the rDNA transcription unit, or sequences downstream of the rDNA transcription unit, increased pTCV1 copy number and stability and conferred additional zones of DNA replication. Sequences upstream of the rDNA transcription unit dramatically destabilized pTCV1. These experiments give additional insights into the mechanism of DNA replication and provide for E. histolytica a set of transfection vectors with unique properties. PMID- 9864857 TI - Actin gene sequence from Euglena gracilis. AB - The full length coding sequence of the Euglena gracilis actin gene was determined by RT-PCR of Euglena gracilis mRNA. Conserved regions in the actin amino acid sequence were used as guides for the synthesis of degenerate primers. Sequence was obtained for 1,238 nucleotides, of which 1,131 were coding for 377 amino acids. Sequence comparisons showed a similarity with other actins of 56% to 80%. Even though most of the actin amino acid sequence was conserved, some regions showed high divergence, i.e. the DNase I-binding loop at the N-terminal region. The construction of a phylogenetic tree based on actin sequences from different organisms placed Euglena gracilis in a cluster with Trypanosoma brucei and Leishmania major. PMID- 9864858 TI - Studies of synaptonemal complexes in farm mammals--a review. AB - For the last 10 years extensive studies of synaptonemal complexes in farm mammals were carried out. In this article a survey of the obtained results is presented. Studies on synaptic behavior of chromosomes at pachytene substage of meiotic prophase I in carriers of centric fusions (cattle, sheep, goat, pig, and blue fox) showed that the trivalent is rarely involved in an association with the sex bivalent. In carriers of reciprocal translocations (pig and cattle) a wide range of synaptic configurations were found. Besides the expected cross-shaped quadrivalent, the following configurations were observed: open quadrivalent, trivalent plus univalent, and two heteromorphic bivalents. The latter configurations were quite frequently involved in the association with XY bivalent. Studies of synaptonemal complexes in interspecies hybrids (cattle x zebu, river x swamp buffalo, blue fox x silver fox) revealed a wide range of synaptic disturbances. Cases of pericentric inversions, aneuploidy, and chromosomal polymorphisms are also reviewed. This survey shows that synaptonemal complex analysis is a very useful tool in studies on causes of altered fertility in carriers of abnormal chromosome complement. PMID- 9864859 TI - Genetic analysis of variation for auxin-induced adventitious root formation among eighteen ecotypes of Arabidopsis thaliana L. Heynh. AB - Eighteen ecotypes and two inbred lines of Arabidopsis thaliana L. Heynh. were analyzed for variation in the number of adventitious roots formed (hereafter referred to as rooting) on seedling hypocotyls in response to auxin treatment. Mean root counts varied from 1.7 to 23.1. Stable high (HA) and low (LA) rooting lines selected from ecotype Columbia, a low rooting ecotype (Mt-0), and unselected Columbia populations were evaluated for vegetative and reproductive growth parameters to determine correlated phenotypic effects of selection for rooting response. High rooting in HA correlated with compact, highly branched shoot growth. Genetic analysis of HA, Mt-0, and their F1, F2, and reciprocal backcross generations indicated that high and low rooting responses in this population may be controlled by several genes acting independently in additive dominance fashion. Genetic variance partitioned into principally additive effects, with dominance favoring low rooting. PMID- 9864860 TI - Physical mapping and characterization of 5S rRNA genes in Douglas-fir. AB - Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] is one of the world's most valuable timber species and is widely used in reforestation. Therefore there is considerable interest in understanding its genetic structure. In conifers approximately 75% of the genome is composed of repetitive DNA. Thus for conifers characterization of repetitive DNA is a significant part of genome analysis. We have characterized the organization of 5S rRNA genes in Douglas-fir at both the molecular and chromosome levels. 5S DNA repeat units containing the coding sequence for 5S rRNA and the nontranscribed spacer (NTS) were cloned using PCR. Sequencing and Southern hybridization revealed repeat units of 888 and 871 bp in length, the latter with a 17 bp deletion in the NTS. The coding region showed high homology with other eukaryotic 5S rRNA genes. A 35 bp region of the NTS immediately upstream of the 5' end of the coding region showed high similarity to other conifers but not to other published plant 5S rDNA sequences. Physical mapping of 5S rDNA by fluorescent in situ hybridization using a biotinilated homologous probe revealed a single subtelomeric site on one pair of large metacentric chromosomes. PMID- 9864861 TI - Genetic aspects of the worldwide colonization process of Ceratitis capitata. AB - Multilocus enzyme electrophoresis data from 26 polymorphic loci (124 alleles) were used to analyze the genetic aspects of the worldwide colonization of Ceratitis capitata (medfly). Eighty-two samples of 17 populations were collected from six regions throughout the species range: Africa, extra-Mediterranean islands (Madeira and Gran Canaria), Mediterranean region, Latin America (Guatemala), Pacific (Hawaii), and Australia. The variability parameters (H, P, A) reveal that the geographical dispersal of medfly from its ancestral source area (East Africa) is associated with a great reduction in variability. The pattern of decreasing variability occurs at two regional levels: in the African Mediterranean region where the differentiation is gradual, and in the Latin American-Pacific region where some ancestral variability is still present as a consequence of recent colonization. The UPGMA phylogenetic tree, derived from Nei's genetic distances, shows the presence of intraspecific differentiative processes affecting mainly the two island populations, Reunion and Hawaii. The population genetic changes observed in the species range are consistent with both the chronology and the historical circuitous course of the medfly colonization process. PMID- 9864863 TI - Hereditary canine spinal muscular atrophy is phenotypically similar but molecularly distinct from human spinal muscular atrophy. AB - Hereditary canine spinal muscular atrophy (HCSMA) is an autosomal dominant motor neuron disease that is similar in pathology and clinical presentation to various forms of human motor neuron disease. We have tested the hypothesis that the canine survival motor neuron (SMN) gene is responsible for HCSMA by genetic and molecular analysis of a colony of mixed breed dogs, all descended from a single affected individual. We cloned the canine SMN gene and determined the DNA sequence in an affected and an unaffected dog. We found no germline mutations in the SMN gene of the affected individual. Using conventional linkage analysis with canine-specific microsatellite repeat markers we screened the canine genome and identified a single linkage group likely to contain the HCSMA gene. Analysis with a panel of canine/rodent hybrid cell lines revealed that the SMN gene did not map to the same chromosome as the HCSMA linkage group. Collectively these results suggest that the molecular basis for HCSMA is distinct from that of phenotypically similar human disorders caused by inherited mutations in the SMN gene. This further suggests that additional studies on the molecular nature of HCSMA may reveal an unknown element of the molecular pathway leading to motor neuron disease. PMID- 9864862 TI - Comparison of acute endotoxin-induced lesions in A/J and C57BL/6J mice. AB - Resistance to the action of endotoxin varies among inbred strains of mice, indicating that a component of this resistance has a genetic basis. Different responses to endotoxin that are characteristic of individual inbred strains represent phenotypes that can be used to genetically map the response modifier genes. This study compares the acute histologic lesions in 8-week-old male A/J and C57BL/6J (B6) mice injected intraperitoneally with endotoxin of E. coli O265:B6 (15 mg/kg). Animals of both strains exhibited splenitis, splenic lymphoid hyperplasia, splenic lymphoid necrosis, and sequestration of neutrophils in the pulmonary alveoli. The B6 mice showed increased margination of white blood cells to the pulmonary vascular endothelium relative to A/J mice. A large number of degenerating neutrophils was observed in the liver sinusoids of most B6 animals, while this lesion was much less severe in A/J mice. This difference was quantified, demonstrating a highly significant difference in neutrophil infiltration in B6 mice relative to A/J mice. Analysis of this phenotype in F1 mice demonstrates that major genes encoding the trait are not X-linked, imprinted, or maternally inherited and do not show the codominant inheritance expected if Lps(d) were primarily responsible. The distinctive, quantitative nature of these differences provides a useful assay for mapping genes that modify endotoxin responsiveness using the AXB and BXA recombinant inbred (RI) strains derived from A/J and B6 mice. PMID- 9864864 TI - Genetic relationships among Japanese native breeds of chicken based on microsatellite DNA polymorphisms. AB - Genetic relationships among Japanese native breeds of chickens were studied on the basis of microsatellite DNA polymorphisms. DNA samples from 10 Japanese native breeds (Iwate-Jidori, Aizu-Jidori, Sadohige-Jidori, Siba-Tori, Onaga-Dori, Echigonankin, Hinai, Kinpa, Koeyoshi, and Tomaru) and one imported breed (White Leghorn) were analyzed using eight microsatellite markers that were isolated from a microsatellite DNA-enriched library of chickens (Takahashi et al. 1996). The PCR primers to detect (CA)n repeat length polymorphisms were synthesized based on the sequences of clones, and these markers were typed by PCR amplification and electrophoresis using a DNA sequencer. Since all eight microsatellite markers were polymorphic, genetic distance between the breeds could be calculated based on the frequencies of alleles of the microsatellites and phylogenetic relationships between the breeds could be estimated. Most Japanese native chickens were grouped into three groups that correspond to the origin breeds, Jidori, Shokoku, and Shamo. The results suggest that microsatellite DNA markers are a useful tool for studying the genetic relationships among chicken breeds. PMID- 9864865 TI - A new allelic series for the underwhite gene on mouse chromosome 15. AB - A new allelic series at the underwhite gene is described. Three of the alleles in the series--uw, uwd, and Uwdbr--arose as spontaneous mutations on different genetic backgrounds at The Jackson Laboratory. We report here the visible phenotypes and dominance hierarchy of these alleles, all of which are defined by a reduction of pigmentation in both eye and coat color. Electron microscopic analysis of retinal epithelium suggests that the primary defect is in the melanosome. The degree of severity of melanosome anomalies in the retina correlates with the degree of hypopigmentation in the coat. The perturbed gene and its gene product are unknown. We show that the uw locus is genetically distinct from Myo10, a suggested candidate gene for this mutation. PMID- 9864868 TI - Elevated serum prostate specific antigen (PSA) related to asymptomatic prostatic inflammation. AB - Prostatic subclinical inflammation is associated with elevated serum prostate specific antigen (PSA) level when there is contact with and disruption of the glandular epithelium of the prostate. This aggressiveness of the inflammation was graded on a 4-point scale proposed by Irani (1). Patients with an elevated serum PSA level and no malignancy in their transrectal prostate biopsy, have a lower risk of biopsy missed prostate carcinoma when there is an inflammation with a high aggressiveness score. The density itself of the inflammatory cells in the prostatic tissue was not associated with the serum PSA level. We believe that both issues should be considered when interpreting a prostate biopsy. PMID- 9864869 TI - Prostate laser operation--the cheapest therapy? 872 cases. AB - Prostate laser operation (PLO) is expensive; but without special fibres--that means f.i. NdYAG- or better Diode-laser--and simple reusable bare fibre it costs only about 20BEF/operation, 872 cases were operated in this way. Bare fibres enable a non-invasive but also invasive coagulation technique as well as an effective laser-cutting. High risk patients and patients under anticoagulation treatment are no contradiction against laser operation. The same laser can be used for many indications in urology--as cheap as in prostate therapy--besides in nearly all medical compartments interdisciplinary especially in smaller hospitals or outpatient therapy. PMID- 9864870 TI - Urodynamics and lower urinary tract symptoms. AB - In a retrospective study four questions concerning urodynamics and lower urinary tract symptoms (LUTS) were answered: Do obstructive symptoms correlate with bladder outlet obstruction? Does the symptom urgency correlate with bladder instability? What is the value of a free uroflow study in men with LUTS? Should all men with LUTS have pressure-flow studies before surgery? The authors state that urodynamic investigation is very important before surgery in patients with LUTS. PMID- 9864871 TI - Non-parasitic chyluria: a case report and review of the literature. AB - Chyluria is due to the passage of chyle into the urine giving it a typical milky appearance. Although rarely seen in our countries it is well-known in areas where infections with Wuchereria bancrofti are endemic. Chyluria can also have non parasitic causes of which only a few cases have been described. We briefly discuss etiologies and pathogenesis of chyluria and present a case of non parasitic chyluria successfully treated by surgery. A short and critical review of the different treatment modalities is made. PMID- 9864872 TI - Bilateral ureteral inverted papilloma with synchronous transitional cell tumor of the bladder. AB - Inverted papillomas are benign urothelial tumors. The majority of cases occur in the bladder, while inverted papillomas of the upper urinary tract are very rare. The association of inverted papillomas of the bladder and urothelial malignancy is documented, but the incidence is low. Inverted papillomas of the upper urothelial tract coexist frequently with urothelial malignancy. Close follow-up with urography and cystoscopy is mandatory in such patients. We describe a case with synchronous bilateral ureteral inverted papilloma and a transitional cell tumor of the bladder, illustrating the strong association of upper urothelial tract inverted papilloma and urothelial malignancy. PMID- 9864873 TI - Vesico-uterine fistula secondary to cesarean section. AB - A vesico-uterine fistula with vaginal urinary incontinence secondary to cesarean section is reported. Diagnosis was made clinically, radiologically and endoscopically. Conservative management failed. Surgical repair was successful. The literature is reviewed, and the treatment options (conservative management- surgical intervention) are discussed. PMID- 9864874 TI - Testicular torsion after previous orchidopexy for undescended testis. AB - We report one case of acute testicular torsion following orchidopexy for an undescended testis. A review of the literature reveals only ten similar cases. History of a previous testicular surgery should not preclude the possibility of a torsion in that testicle. We conclude that at orchidopexy for an undescended testis, eversion of the tunica vaginalis is an essential step to avoid any future torsion. PMID- 9864875 TI - [Testicular metastasis of transitional cell carcinoma of the prostate]. AB - This case illustrates in a 55 year old man a testicular metastasis from a prostatic TCC occurring fourteen years after a bladder localisation of the same type. To our knowledge no other similar observation has been reported in the literature so far. Metastatic disease should be considered in the differential diagnosis of testis tumours arising in patients with a history of urothelial TCC from bladder, ureter or even prostate as described here. PMID- 9864876 TI - A cystine lower pole renal calculus treated with holmium: YAG laser using a flexible 9,5F transurethral ureteroscope. AB - We report a case of a 1.5 cm cystine staghorn calculus of the right lower pole in a 32 year female known cystinuric patient. With a 200 microns Holmium laser probe through a 9,5 F flexible ureteroscope the calculus was fragmented in small particles. An internal ureteral stent was inserted at the end of the procedure. All but one small residual fragments were evacuated spontaneously after removal of the stent. This case shows that flexible ureteroscopy combined with the Holmium laser is a safe and efficient procedure to treat medium size renal cystine calculi. It can be repeated in case of recurrence with minimal trauma to the urologic tract. PMID- 9864877 TI - Ureteroscopic treatment of ureteric lithiasis. Analysis of 354 urs procedures in a community hospital. AB - INTRODUCTION: The optimal treatment of ureteric lithiasis continues to be a point of discussion, as there are different treatment modalities. Conservative treatment, EWSL, ureteroscopy (URS), percutaneous litholapaxy, and (laparoscopic) ureterolithotomy, all have there place in the treatment of ureteric lithiasis. In order to evaluate if ureteroscopy is a valuable alternative to ESWL in the treatment of ureteric lithiasis, an analysis of all the ureteroscopies performed in our institution was made and these results were compared with results (both ESWL and URS) described in literature. PATIENTS AND METHODS: During the period 1990 till 1997, 292 patients underwent in our institution in total 354 ureteroscopies for ureteric lithiasis. In all procedures a rigid 9 Fr. ureteroscope was used, together with laser lithotripsy or pneumatic lithotripsy as fragmentation device. RESULTS: Overall success rate in our series was 90.1%, with distal and middle ureteric stone location being more favorable (94% and 95%) than proximal location (73%). Overall complication rate was 7.6% (including minor complications, such as ureteral mucosal tear), for which open surgical intervention had to be performed in 2.3% of cases. CONCLUSIONS: Although invasive, ureteroscopy proved to be a very competitive alternative to ESWL, when treating ureteric lithiasis. Success rates are equal, if not better, using ureteroscopy when compared to ESWL. Complications of ureteroscopy are infrequent, but do exist. Therefore ureteroscopy should be performed by experienced urologists. PMID- 9864878 TI - Outbreak of echovirus 30 meningitis in southern Saskatchewan. PMID- 9864879 TI - Aseptic meningitis due to echovirus 30, Japan, 1997-1998. PMID- 9864880 TI - Outbreak of quinolone-resistant, multiresistant Salmonella Typhimurium DT104, Denmark. PMID- 9864881 TI - [Role of the endometrium in human embryonic implantation]. PMID- 9864882 TI - [Is there a role for GnRH analogs in ovulation stimulation for in vitro fertilization?]. PMID- 9864883 TI - [Role of hysteroscopic treatment of the septate uterus resulting in primary sterility]. PMID- 9864884 TI - [Prevention of prematurity by psychotherapeutic management]. AB - The establishment of the mother-baby relationship is often disturbed in case of preterm birth. Clinical observations and epidemiological studies have highlighted the existence of a relationship between psychological factors and preterm birth. Therefore, a psychological support may have a beneficial effect in order to prevent preterm delivery. The aim of this study was to assess the effectiveness of psychological support in pregnant women with symptoms of preterm labor, which is considered as a somatic expression of psychological difficulty related to pregnancy. The study, which involved two cohorts of women, hospitalized or not, identified during two successive periods in the same maternity ward, included 309 women in the experimental group and 323 in the control group. The women in the control group were followed up according to the usual therapeutic procedures, whereas additional psychological support was offered to the experimental group. 92% of the women who met the psychologist accepted the psychological support. The analysis, conducted "in intent to treat", showed a significant decrease in the preterm birth rate in the experimental group (12%) compared with the control group (26%). This study confirms the feasibility of this kind of intervention and the effectiveness of psychological support on the risk of preterm delivery. The interviews help the women regain confidence in their own capacity to fulfil their pregnancy. PMID- 9864885 TI - [Atomic absorption spectrophotometry study of copper ion release by copper bearing intrauterine devices]. AB - Copper release from copper-bearing IUD's was studied in vitro and in vivo using atomic absorption spectrophotometry in deionized water, normal saline solution and normal ovulatory cervical mucus. In these media, copper release from a 375 mm2 DIU occurs without latency, showing comparable amounts for identical time intervals. Daily copper release was shown to be respectively 8 and 11 times higher in cervical mucus and normal saline solution than in deionized water. Although copper ions are detectable in ovulatory cervical mucus under physiological conditions, the copper content appears 5 to 6 times higher in women bearing a copper IUD. Obviously, the copper amount is dependent on the copper exposed surface: the daily in vitro release from a 250 mm2 IUD is 18% inferior to that observed from a 375 mm2 model. In vivo, the daily copper release in ovulatory mucus of 380 or 200 mm2 IUD users is respectively 5 and 3.5 times higher than in controls. PMID- 9864886 TI - [Outcome of patients treated with intra-peritoneal immunochemotherapy for ovarian cancer]. AB - OBJECTIVE: The aim of this study is to show that i.p. chemotherapy improves the evolution of the patients with an ovarian cancer, as the risk of a small number of complications due to this route of administration. METHODS: 85 patients (11 stage Ic, 4 IIc and 70 stage III) were treated from 1980 to 1993 (median of follow up > 77 months) by a cisplatin based immunochemotherapy administered intraperitoneally, by a needle. RESULTS: Median overall survival exceeds, 72 months (60 months for stage III). Out of 41 relapses, 11 took place after 48 months of follow up. Delayed general complications consisted of 2 chronic nephotoxicities due to CDDP and one leukemia. Local complications are dominated by serous adherences (5 cases of plastic peritonitis). Secondary cytoreductions turned out to be beneficial in this series in spite of relapse diagnosis of them being carried out to late. CONCLUSION: Classic imagery remains disappointing for the exploration of the pelvic abdominal region. Paclitaxel by i.p. way seems very promising, which should launch once again interest in secondary debulking surgery. PMID- 9864887 TI - [Natural history of ovarian adenocarcinomas: from epidemiology to experimentation]. AB - The ovarian carcinogenesis is not well known because the diagnosis is usually delayed to an advanced stage with a lot of chromosomic abnormalities. On the other hand, there is a lack of animal models and preneoplastic patterns are discussed. Different ways of studying are in progress. Prophylactic oophorectomy in women with BRCA1 mutation provides ovaries that can be compared to a control group. Epithelial or stromal changes are observed in the familial risk group more frequently than in the controls and could be preneoplastic. An experimental approach has been developed in order to confirm the Fathalla's theory of the incessant ovulation by culturing cells of rat surface ovarian epithelium with a high proliferative pressure. These cells become malignant in 10 out of 30 cases (after 10 cultures). The transformed tumor cells are a syngeneic tumor model that can be transplanted in the animal and analysed by molecular biology technics. This allowed to show the loss of a potentially tumor suppressor gene, lot 1, in the transformed cells and on the other hand the overexpression of a gene coding for a protease enzyme the cathepsin B, which is a marker of an aggressive metastatic power. PMID- 9864888 TI - [Taxanes in the treatment of breast and ovarian cancers: current indications and results]. AB - Since the last years, taxanes are among the most active molecules in the treatment of advanced breast and ovarian cancers. Concerning the ovarian cancer, Paclitaxel in association with Cisplatinum is about to become the first line reference protocol. Studies are going on in order to define the association and the optimal way of administration. Concerning breast cancer, Docetaxel seems to provide the most interesting response rates after failures of anthracyclines. In France, randomised studies in adjuvant situation have started with this molecule. PMID- 9864889 TI - [Questions about sexual ethics related to AIDS]. AB - The discovery of Aids has profoundly interfered with the sexual behaviour, perceptions and discourse. It has provoked new questions in the field of sexual ethics, some of which are presented and discussed here: safer sex, partner notification, child wish and the attitude of society towards Aids. Finally a discussion on these questions at large is presented. A responsibilisation of each person concerned, within a context of respect for his/her individual autonomy and a positive attitude towards sex and pleasure, is essential. This responsibilisation may be realised in different ways, aimed at different groups in society: there are campaigns for a large public on the one hand and "targeted prevention" on the other. The role of society is to create socio-economic conditions and to take actions promoting responsible behaviour by all its members. PMID- 9864890 TI - [Circulating antibodies against an epitope of the heat-shock protein 60kDA of chlamydia trachomatis in infertile couples. Relation with surface chlamydia trachomatis anti-antigen antibody and E. coli anti-HSP60 antibody and human anti HSP60]. PMID- 9864891 TI - [Lymphatic drainage in stage IB and II cervical cancers: anatomical study and surgical deductions]. PMID- 9864892 TI - [Carotid stenosis--noninvasive diagnosis preferred. Suitability of color-coded duplex ultrasound for preoperative diagnosis]. AB - The high incidence of cerebral ischemic insults caused by atherosclerotic lesions in the carotid artery warrants a careful diagnostic work-up if the therapeutic possibilities now available are to utilized to the full. Until recently, the diagnosis of carotid stenosis was considered the domain of arterial angiography, but MRI angiography and, in particular, color-coded duplex ultrasonography, have now made these techniques so reliable that-as this case report shows--there is no longer any need for routine angiography for surgical planning. Furthermore, even arteriography involving standard projections may fail to provide the necessary information. PMID- 9864894 TI - [Costs of intensive insulin therapy in type 1 diabetes mellitus. Experiences from the DCCT study]. AB - Diabetes mellitus is one of the most common metabolic diseases in many countries of the world. Its prevalence in Germany has increased 7- to 8-fold over the past 30 years. The clinical and economical importance of diabetes is determined by the frequent occurrences of such serious complications as neuropathy, retinopathy and nephropathy. Intensive insulin therapy with regular monitoring of blood glucose (up to 4 measurements daily) and adjustment of the insulin dose accordingly may achieve virtually normal levels of blood glucose and thus decrease the risk of these complications. The present cost-effectiveness-study shows that the higher costs of invasive insulin therapy are offset by savings of 8.114 German marks per patient resulting from the reduction in morbidity and mortality. On the basis of an estimated 5% to 10% type 1 diabetes among the total diabetic population (prevalence 4.9%), potential saving of 1.62 to 3.24 billion marks are calculated for Germany. PMID- 9864893 TI - [Current insulin therapy--avoidable errors]. AB - Since its introduction in 1922, insulin therapy has undergone constant development and improvement. Today, the spectrum ranges from short- to very long acting depot insulins, and "human" insulins. Further stages in the development of this form of treatment have been intensive insulin therapy, self-measurement of blood sugar, development of insulin-analogues and the use of insulin pumps or pens, to mention but a few. As a reflection of how widespread insulin therapy in the doctor's office has become and what the frontiers are, the present paper lists a range of 15 errors, and discusses how they can be avoided and insulin treatment thus further optimized. PMID- 9864895 TI - [Rheumatoid arthritis. 2: Differential diagnosis and therapy]. PMID- 9864896 TI - [Investigation of the impact of environmental risks on health and its control]. PMID- 9864897 TI - [Do chronic diseases prevent intra-hospital mortality? Paradoxes and biases in information about hospital morbidity]. AB - OBJECTIVE: Previous studies have demonstrated how the incomplete codification of the secondary diagnostics can bias the estimation of the risk of in-hospital death based on clinical-administrative databases. The objective of this study is to measure the trend of the association between in-hospital mortality and the secondary diagnostics register in the Minimum Basic Data Set (MBDS) of the Valencian Community. METHODS: The 14,161 admissions of persons over the age of 64 were extracted from the MBDS (years 1993-94) for: stroke (S), bacterial pneumonia (BN), myocardial infarction (MI) and congestive heart failure (CHF). The relation was measured between the availability of some additional diagnostics (selected to dispose of a group of heterogeneous chronic and acute processes), and the risk of in-hospital death, relative risk (RR) and adjusted odds ratios (aOR) were calculated per age, gender, length of stay and number of diagnoses. RESULTS: Many of the conditions are associated with a reduced risk of death such as the diabetes mellitus (Mortality for stroke, RR: 0.58; aOR: 0.53), old myocardial infarction (mortality for myocardial infarction, RR: 0.40; aOR: 0.35) or hypertension (mortality for stroke, RR: 0.54; aOR: 0.49): this also occurs in angina, coronary atherosclerosis, aortic and mitral valve disease, atrial fibrillation, chronic obstructive pulmonary disease and urinary tract infection. CONCLUSIONS: Although there maybe other explanations, such as the existence of a bias in the hospital selection, the explanation which is most in agreement with the findings is the presence of an information bias in the MBDS due to the tendency to register less information about chronic antecedents of the persons who die. PMID- 9864898 TI - [Association of air pollution caused by sulphur dioxide and total number of particles in suspension and daily mortality in the city of Madrid (1986-1992)]. AB - OBJECTIVE: The aim of the study is to quantify the relationship between sulphur dioxide (SO2), total suspended particulate (TSP) and daily mortality in Madrid. METHODS: Data were plotted to exhibit the functional relationship between SO2, TSO and mortality. Box-Jenkins prewhitening method was used to detect lags and weights from the transfer function. Multivariate ARIMA models were built to control for confounding variables (air temperature and influenza epidemics). RESULTS: Graphic analysis suggested a logarithmic relationship between SO2 and mortality, and lineal relationship with TSP. No evidence of a threshold was found for each pollutant. Cross-correlation functions showed a statistically significant relationship between daily mortality caused by organic, cardiovascular and respiratory diseases and air pollution. The first lag was statistically significant for TSP and the third lag for SO2. Multivariate models gave a coefficient beta = 0.039 for TSP and beta = 1.04 for SO2 indicating a 6.6% increase in mortality for each 100 micrograms/m3 increase in TSP and a 2.2% for SO2. CONCLUSIONS: The result suggest a causal relationship between daily mortality and air pollution in Madrid. PMID- 9864899 TI - [Lead blood levels in children in the metropolitan area of Madrid]. AB - The objective is to determine the blood lead levels of the children (7-8 years of age) living in some of Madrid's satellite towns, and to establish links between these levels and different socioeconomic items, life styles and environmental factors, among other variables. A cross-sectional study using a sample based on 205 schoolchildren of Second Grade Primary Education, was under taken during 1995. Venous blood samples were analysed through atomic absorption spectrophotometry. Parents answered a questionnaire with items used in this study. The average blood lead level found was 3.9 mud/dl (CI 95% 3.7-4.1), with higher levels among the children of the Northern zone of Madrid. Between the associated factors to blood lead level was found to be the habit of nail biting, mother's studies and some variables associated with habitual dwelling. We reached the conclusion that the blood lead levels of Madrid's suburban children are low. PMID- 9864900 TI - [Impact of air pollution on the mortality and emergencies of chronic obstructive pulmonary disease and asthma in Barcelona]. AB - OBJECTIVES: Air pollution has been associated with increased mortality according to studies carried out in the US. The APHEA project (Air Pollution on Health: a European Approach) analyzes the short-term effects in 15 european cities. We evaluated the acute relation between air pollution, mortality, and hospital emergency-room visits in Barcelona, one of the cities participating in the APHEA project. METHODS: Daily variations in total mortality, cardiovascular mortality, respiratory mortality, and emergency-room visits for chronic obstructive pulmonary disease (COPD), and asthma were studied in relation to daily variations in air pollution levels in 1985-1991. Poisson regression was done and temperature, relative humidity, and epidemics of asthma and flu were controlled. Temporal trends and auto-regressive terms were examined. RESULTS: A reduction of about 50 micrograms/m3 in particles and sulfur dioxide was accompanied by a reduction of about 4% and 6% (p < 0.05), respectively, in daily deaths from respiratory and cardiovascular causes and emergency-room visits for COPD. Oxidant pollutants (nitrogen dioxide and ozone) were related positively with cardiovascular mortality and emergency visits for COPD and asthma. The role of ozone was notable, with a reduction in ozone levels of 50 micrograms/m3 originating a 4% reduction in emergency-room visits for COPD and asthma (p < 0.05). CONCLUSIONS: Current levels of air pollutants had an epidemiologically measurable impact on mortality and emergency-room visits in Barcelona. These results were consistent with the findings of similar studies in other european and american cities and with previous studies of emergency-room admissions in Barcelona. These studies suggest the possible toxicity of air pollution. PMID- 9864901 TI - [Audit of a hospital internal communication]. AB - OBJECTIVES: To know the current status of internal communication as perceived by the professional staff (physicians, nurses), middle-level managers, and executive team, and possible future strategies for improving internal communication as perceived by the professional staff. METHODS: The study approach consisted of qualitative methods such as focal groups and semistructured interviews. The study area was a regional hospital. Study subjects included the attending staff (physicians and nurses), middle-level managers (supervisors and department heads), and executives (area heads, nursing director, and medical director). From the base staff, 20 nurses and 18 physicians were chosen for four focal groups. Among the middle-level managers, 2 supervisors and 4 department heads were chosen randomly. Three executive officers were interviewed. RESULTS: The weak and strong points of the hospital's internal communication were shown. Nursing and medical staff noted as an important weak point the fact that they did not feel that directors were listening to them. The professional staff noted as a weak point that they do not receive congratulations or positive reinforcement of any type. With respect of downward information channels, the staff indicated that they do not work because their first information received is always in the form of rumors. The physicians evaluated as a strong point that information always passes through the department head. They evaluated as a strong point good intradepartmental relations. The nurses evaluated as a weak point the obstacles to communication that they encounter. They propose future improvements such as developing itinerant communication, electing directors, using informal channels, recognizing staff achievements, and developing a communication plan. CONCLUSIONS: Internal communication should be understood by executives as part of basic professional management strategy. Given the results of this audit, it will be important to establish an internal communication plan to advance hospital objectives in every sense, at every level and for all professional categories. PMID- 9864902 TI - 51st Annual meeting of the Gerontological Society of America, Philadelphia, Pennsylvania, USA. November 20-24, 1998. Abstracts. PMID- 9864903 TI - [Computer-assisted analysis of cardiotocographic recordings in the course of fetal hemolytic disease]. AB - OBJECTIVES: The aim of this study was to assess the usefulness of computerized cardiotocography in the diagnostics of fetal hemolytic disease. STUDY DESIGN: 50 invasive diagnostic and therapeutic procedures proceeded by computer assisted CTG recording were analysed. The values of particular parameters of CTG were compared with adequate values obtained in uncomplicated pregnancy. The correlation between both cardiotocographic and hematological parameters was estimated. The common relationship between all analyzed features was examined using multidimensional analysis of variancy. RESULTS: The most significant correlation was determined in case of short-term variation and hematocrite of fetal blood. The five-feature algorhythm, containing short- and long-term variation, number of accelerations, fetal movements and gestational age, was established for the prognosing of the most probable level of fetal hematocrite. CONCLUSION: Computerized cardiotocography seems to be the useful diagnostic method which enable to decrease the number of invasive intrauterine procedures in case of fetal hemolytic disease. PMID- 9864904 TI - [Transvaginal color Doppler study of the uteroplacental circulation in early pregnancy]. AB - OBJECTIVE: To compare resistance index (RI) and pulsatility index (PI) values of blood flow in arteries of uteroplacental circulation in normal and threatened abortion first trimester pregnancies. STUDY DESIGN: We have analysed 36 pregnant woman between 5th and 12th weeks of pregnancy--17 patients with threatened abortion as a test group and 19 patients with normal intrauterine pregnancy taken as controls. In all patients transvaginal ultrasonography with pulse color Doppler was performed. The RI and PI values were calculated for blood flow velocity waveforms obtained from uterine artery and its branches--arcuate, radial and spiral arteries. RESULTS: In the test group we have analysed 17 flow velocity waveforms from uterine artery (mean RI 0.852; SD 0.080, mean PI 2.324; SD 0.693), 16 from arcuate artery (mean RI 0.638; SD 0.049, mean PI 1.315; SD 0.064,), 17 from radial artery (mean RI 0.638; SD 0.049, mean PI 1.009; SD 0.177) and 15 from spiral artery (mean RI 0.548; SD 0.068, mean PI 0.804; SD 0.160); in controls we have analysed 16 flow velocity waveforms from uterine artery (mean RI 0.866; SD 0.066, mean PI 2.469; SD 0.618), 17 from arcuate artery (mean RI 0.728; SD 0.123, mean PI 1.352; SD 0.362), 19 from radial artery (mean RI 0.630; SD 0.092, mean PI 1.017; SD 0.261) and 16 from spiral artery (mean RI 0.511; SD 0.100, mean PI 0.760; SD 0.202). CONCLUSION: There was no significant difference in mean RI and PI values between normal and abnormal pregnancies. A gradual statistically significant decrease of RI and PI values during time of pregnancy we have observed only in radial arteries in both groups. For other vessels in the control group the RI and PI values tended to decrease during the time of pregnancy while in the test group some even rose up. PMID- 9864905 TI - [Obstetric hysterectomy]. AB - An analysis of indications for obstetric hysterectomy in Central Teaching Hospital of Ministry of Internal Affairs in Warsaw between 1985-1997 is presented. There have been 15 caesarean or post partum hysterectomies over that period, which makes 0.11% of all deliveries and 0.67% of caesarean sections. All operations followed caesarean sections. In two patients (13.3%) procedure was performed electively because of myomas. In the remainder emergency or post-partum hysterectomy was necessary. The most common indication was haemorrhage connected with placenta praevia, adherent placenta and abruption of placenta (53.3%). Other indications included ruptured uterus (6.7%), severe postpartum or intrapartum haemorrhage due to uterine atony (20.0%) and HELLP syndrome (6.7%). CONCLUSION: 1. Complications associated with pathologies of placenta are the most common indication for obstetric hysterectomy in our study. 2. Patients that are mostly exposed to the risk of obstetric hysterectomy are multiparous women especially with previous caesarean section. 3. The incidence of obstetric hysterectomy seems to be rising, possibly as a consequence of the rising rate of caesarean section and uterine curretage. PMID- 9864906 TI - [Acid-base balance in hypotrophic neonates in the first hours of life]. AB - Our purpose was to present parameters of acid-base status at 30, 90 and 150 minutes after delivery in cases of hypotrophic infants. Significantly lowered values of pH, HCO3 and BE were found as compared to results of normal term infants. Parameters are consistent with metabolic acidosis. Such conditions require appropriate treatment. PMID- 9864907 TI - [The evaluation of concentration myeloperoxidase and interleukin-8 in blood serum of pregnant women with or without pregnancy induced hypertension]. AB - In the serum of woman during the pregnancy with and without PIH the activity of interleukin-8 and myeloperoxydase was measured. 40 pregnant woman with PIH and 16 normal pregnant woman were included in the study. We have not found significative difference between examined populations. PMID- 9864908 TI - [The course of twin pregnancy and delivery in a woman after mitral valve implantation]. AB - We described the course and the way of twin delivery of patient, who was pregnant one month after implantation of mitral heart valve. We discussed the threats for the fetus and mother, connected with administration of anticoagulant drugs. PMID- 9864909 TI - [Malignant teratoma of the sacrococcygeal area in a fetus unrecognized in pregnancy]. AB - A very rare case is presented of a giant teratoma malignum of the sacrococcygeal area in a fetus which caused obstetric difficulties. Attention in called to the necessity of differentiating such teratoma from other tumors in the region of the body, which is important for the decision concerning further prognosis and the time and mode of pregnancy termination. Good prognosis in cases of teratoma in this area suggests termination of pregnancy by ceasarean section. The newborn with teratoma was referred on the second day of life to surgical department where the tumour was removed radically without damaging the nearly organs. The need for more accurate prenatal diagnosis and labour management in specialised centres with available facilities for paediatric surgery are suggested. PMID- 9864910 TI - [A long-time presence of a foreign body in an episiotomy scar]. AB - OBJECTIVES: The object of our study was a long-time presence of a foreign body in an episiotomy scar. STUDY DESIGN: A case of 46-year old woman with a palpable tumour in a perineal region is described. RESULTS: This false tumour was caused by a surgical needle which was left in the body of the patient 19 years ago during the episiorrhaphy after the last labour. CONCLUSION: Because of a character and localisation of the change the method of choice was surgical treatment which completely eliminated all symptoms. PMID- 9864911 TI - [Hormonal dependence of vesicouterine fistulas]. AB - Vesicouterine fistulas are more and more frequently reported. This paper reviews available data suggesting the role of the endometrium in the fistula's formation. At least a proportion of these fistulas fulfil the criteria of external endometriosis. Conservative management by means of hormonal treatment should be considered before surgical repair. We also present a new clinical classification of this disorder, based on the routes of menstrual flow: type I--with menouria, type II--with dual flow via both the bladder and vagina, and type III--with normal vaginal menses. PMID- 9864912 TI - Prophylactic platelet transfusion in acute leukemia: which threshold should be used. PMID- 9864913 TI - The role of serum transferrin receptor in the diagnosis of iron deficiency. AB - BACKGROUND AND OBJECTIVE: Iron deficiency anemia (IDA) is often associated with inflammatory disorders. The most conventional parameters of iron metabolism are therefore affected, making the evaluation of iron status difficult. Serum transferrin receptor (sTfR) levels are raised in iron deficiency but are not influenced by inflammatory changes. The aim of this study was to investigate the role of sTfR in differentiating IDA with inflammatory features. DESIGN AND METHODS: A diagnostic study of sTfR measured by immunoassay was carried out in IDA and anemia of chronic disorders (ACD). The cut-off points of sTfR and the ratio of sTfR/serum ferritin, which were obtained after comparing IDA and ACD, were applied to a group of 64 patients with mixed iron patterns (MIX) (16 with ACD and 48 with IDA). RESULTS: The best cut-off point of sTfR between IDA and ACD was 4.7 mg/L. Applying this cut-off to the MIX group, an efficiency of 87% was obtained (sensitivity 92% and specificity 81%). This level of sTfR correctly classified 53 out of 64 cases of the MIX group (83%). Using the ratio of sTfRx 100/serum ferritin, the best cut-off point was 8 (efficiency 100%), which correctly classified 62 out of 64 cases of the MIX group (97%). INTERPRETATION AND CONCLUSIONS: This study demonstrates that sTfR in conjunction with other iron parameters is very useful in iron deficiency evaluation, especially in hospital practice. Iron treatment should be considered in patients with mixed patterns of iron status, in which the diagnosis of IDA versus ACD is difficult, when the levels of sTfR exceed the cut-off point. PMID- 9864914 TI - Clinical relevance of CD10 expression in childhood ALL. The Italian Association for Pediatric Hematology and Oncology (AIEOP). AB - BACKGROUND AND OBJECTIVE: Previous studies have considered the prognostic significance of CD10 expression in childhood acute lymphoblastic leukemia (ALL) and showed its linkage to a more favorable prognosis. The aim of this study was to assess the independent significance of CD10 expression in a large population of ALL patients. DESIGN AND METHODS: We revised the independent clinical relevance of CD10 expression in 2038 children with acute lymphoblastic leukemia (ALL), who were consecutively entered in 4 sequential trials of the Italian Association for Pediatric Hematology and Oncology (i.e. AIEOP studies 82, 87, 88, 91); 1142 were males and 896 females, age ranged between 1 and 14 years (yrs) at diagnosis. Of the whole group, 1471 children (72.2%) were defined as having standard risk, 567 (27.8%) as having a high risk. RESULTS: CD10 was detected in blast cells from 1706 of 1784 (95.6%) patients with B-lineage ALL and 46 of 254 (18.1%) with T-cell ALL. In the B-lineage subgroup CD10 expression was associated with presenting features such as age < 9 yrs and inclusion in the standard risk category. No significant differences were found between CD10+ and CD10- cases in T-lineage ALL, concerning presenting features, except for FAB L2 in the former group. We compared the event-free survival (EFS) rates for patients with T-ALL or B-lineage ALL, regarding CD10 positivity, overall and by individual study. Patients with T-ALL fared worse than those with B-lineage ALL (5 and 10 yrs EFS: 46.8% vs. 68.5% and 44.5% vs. 63.7% respectively, p = 0.0001). In multivariate analysis of B-lineage subgroup poorer EFS was associated with male sex, higher WBC (> or = 20 x 10(9)/L), age > 9 yrs. Only WBC > or = 20 x 10(9)/L and age > 9 yrs were parameters linked to poorer EFS in the T-lineage subgroup. Finally, we compared EFS rates for four groups of patients categorized as having high or standard risk, and according to CD10+ and CD10- expression. High-risk patients fared statistically worse than standard risk patients both in the CD10- and in the CD10+ groups (42% vs. 50.7% and 63.6% vs. 66.8%, respectively). INTERPRETATION AND CONCLUSIONS: CD10 expression does not have independent prognostic significance in either the larger subgroup of B-ALL patients or in T cell ALL. PMID- 9864915 TI - The K562 chronic myeloid leukemia cell line undergoes apoptosis in response to interferon-alpha. AB - BACKGROUND AND OBJECTIVE: The K562 cell line, derived from a chronic myeloid leukemia (CML) patient and expressing B3A2 bcr-abl hybrid gene, is known to be particularly resistant to apoptotic death. IFN-alpha treatment of CML patients impairs malignant cell clone, apparently protecting from progression to terminal blast crisis. The mechanisms underlying this kind of cell deletion are analyzed here by multiple technical approaches. DESIGN AND METHODS: K562 cells, variably treated with IFN-alpha, were examined by agarose gel DNA electrophoresis, light and electron microscopy. The presence of bcr-abl rearrangement was revealed by RT PCR. RESULTS: At 4 day treatment both DNA ladder and apoptotic nuclear changes were identified, consistently in the presence of bcr-abl expression. INTERPRETATION AND CONCLUSIONS: Even cells expressing bcr-abl, such as K562, can be triggered to apoptosis. Therefore, this genetic condition, commonly preventing PCD, does not prevent IFN-alpha-mediated apoptosis. PCD seems thus to be the mechanism underlying IFN-alpha-treated K562 cell deletion and it could be the basis of malignant clone reduction in IFN-alpha treated CML patients. PMID- 9864916 TI - Treatment of Ph1-positive chronic myelogenous leukemia in children: comparison between allogeneic bone marrow transplantation and conventional chemotherapy. Spanish Working Party for BMT in Children (GETMON). AB - BACKGROUND AND OBJECTIVE: To compare the estimated survival and disease-free survival between children with Ph1-positive chronic myeloid leukemia treated with allogeneic bone marrow transplantation or conventional chemotherapy. DESIGN AND METHODS: In this retrospective study we compared the results obtained in a group of 14 children who received allogeneic bone marrow transplantation (BMT) between 1983 and 1993, and another group of 27 children treated with busulfan, hydroxyurea or alpha-interferon during the same time period. Patients were transplanted at a median of 7 months from diagnosis and all except one were in their first chronic phase. Conditioning consisted in total body irradiation and cyclophosphamide in 12 cases, and busulfan was added in two. RESULTS: Of the 14 patients treated with BMT, two died of transplant-related complications and two relapsed 18 and 48 months after the BMT. Ten children remain alive and disease free at a median follow up of 60 months. The probability of DFS at 5 years is 70%. Of the 27 patients treated with chemotherapy, 22 have died at a median of 36 months from diagnosis. The probability of survival at 5 years is 5% versus 83% for the BMT group (p = 0.001). INTERPRETATION AND CONCLUSIONS: Allogeneic BMT is a safe and very effective treatment for Ph-positive CML in children. Patients who have an HLA-identical sibling donor must receive a transplant as soon as possible after being diagnosed. PMID- 9864917 TI - Disappearance of PML/RAR alpha acute promyelocytic leukemia-associated transcript during consolidation chemotherapy. AB - BACKGROUND AND OBJECTIVE: Acute promyelocytic leukemia (APL) (M3 according to FAB classification) is a subtype of acute myelogenous leukemia characterized by a specific t(15;17) (q22;q12) chromosomal translocation. The majority of APL patients achieve morphologic remission after induction chemotherapy. They can be followed from this point by cytogenetic and molecular analysis of the persistence of the PML/RAR alpha transcript. In order to determine the influence of successive courses of consolidation chemotherapy on clinical and molecular outcome, APL patients treated with all-trans retinoic acid (ATRA) and chemotherapy (AIDA-GIMEMA-LAP0493 protocol) were investigated. DESIGN AND METHODS: Twenty-four APL patients (pts) (15 males; 9 females) were studied by RT PCR and cytogenetic analysis at diagnosis, after induction chemotherapy, at each point after any of three consolidation courses, and every 3 months during the first years of maintenance therapy. The median follow-up was 24 months (mths) (range 7-40 mths). RESULTS: All pts achieved hematologic remission after induction chemotherapy. Our results demonstrate that the majority (87%) of APL patients were still molecularly positive for the APL associated transcript after induction chemotherapy, while the majority (80%) of APL patients became PCR-after the second consolidation chemotherapy. INTERPRETATION AND CONCLUSIONS: The role of the third consolidation chemotherapy course in converting patients with persistent molecular evidence of disease from PCR+ to PCR- was minimal. We confirm the validity of molecular follow-up after single courses of chemotherapy in monitoring the role of molecular remission. PMID- 9864918 TI - Ultrasound-guided core-needle biopsy is effective in the initial diagnosis of lymphoma patients. AB - BACKGROUND AND OBJECTIVE: With the development and refinement of new guidance methods for percutaneous biopsies, many investigators have reported studies supporting a role for radiologically guided core-needle biopsy in the diagnosis of malignant lymphoma under certain clinical circumstances. The aims of this report are to evaluate the efficacy of findings at ultrasound (US)-guided core needle biopsy of abdominal lymphoma on patient care and define the key determinants of clinical success. DESIGN AND METHODS: US-guided core needle biopsies were performed in 55 patients with abdominal lymphoma: 44 non-Hodgkin's lymphoma (NHL) and 11 Hodgkin's disease (HD); 41 had had no prior lymphoma and 14 had previously diagnosed lymphoma. All the biopsies were performed under US control using a 21-gauge modified Menghini needle. Overall, 53/55 (96%) patients were treated on the basis of biopsy findings only, including 14/14 (100%) patients with a history of lymphoma and 39/41 (93%) patients with no such history. RESULTS: In 46/53 (87%) patients it was possible to assess the specific histotype. No differences between the diagnostic rates of HD and high grade-NHL were recorded. There were no complications related to the biopsies. INTERPRETATION AND CONCLUSIONS: Our data indicate that abdominal US-guided core needle biopsy should be considered as an effective and safe procedure in the diagnosis of patients with lymphoma offering the possibility of determining the tumor subtype and the subsequent specific treatment. PMID- 9864919 TI - Retrospective analysis of mantle cell lymphoma: experience of the "Gruppo Italiano per lo Studio dei Linfomi" (GISL). AB - BACKGROUND AND OBJECTIVE: Mantle cell lymphoma is a recently recognized histologic entity with specific biological and clinical features. Clinically, the reported unfavorable outcome of these patients has focused attention on this category of non-Hodgkin's lymphoma (NHL). DESIGN AND METHODS: The slide specimens of 69 NHL patients, originally classified as Working Formulation (WF) group B and E, were reviewed. The clinical features at presentation, response to therapy, response duration and survival were analyzed in cases reclassified as MCL. The correlation between clinical and histologic characteristics and the final outcome was evaluated. RESULTS: Out of 69 cases, 34 specimens were reclassified as MCL; in 6 patients, previously classified as WF group B, the nodular pattern was confirmed; in 2 instances the blastoid form was recognized. After a median follow up of 35.7 months, the entire series displayed a median overall survival of 41.2 months; a significantly longer survival was associated with the nodular histologic pattern, IPI score < 2, response achievement, and a higher Hb level. The vast majority of patients received anthracycline-containing combination chemotherapy. Complete remission rate was 38.8% and overall response rate was 67.6%; response achievement was significantly influenced only by Hb level. Median response duration was 23.3 months. INTERPRETATION AND CONCLUSIONS: The present study confirms the unfavorable clinical course of MCL and the possible need for an alternative therapeutic strategy for this NHL category. Therefore, the correct identification of MCL at diagnosis appears of relevance. PMID- 9864920 TI - Prophylactic platelet transfusion threshold during therapy for adult acute myeloid leukemia: 10,000/microL versus 20,000/microL. AB - BACKGROUND AND OBJECTIVE: The threshold for prophylactic platelet transfusions has been classically established at 20,000/microL. In 48 patients with de novo acute myeloblastic leukemia (AML) we analyzed the effect of reducing the threshold for prophylactic platelet transfusion from 20,000/microL (group A) to 10,000/microL (group B) after induction and consolidation chemotherapy. DESIGN AND METHODS: Forty-eight adult patients with de novo AML diagnosed in a single institution in a nine year period were enrolled in the study. Between January 1989 and December 1993 the patients received prophylactic platelet transfusions when their platelet count was below 20,000/microL (group A), and from January 1994 to March 1998 prophylactic platelet transfusions were indicated below 10,000/microL or between 10,000/microL and 20,000/microL if there was any consumption factor. RESULTS: The mean number (SD) of platelet transfusions during induction was 8.4 (5.3) in group A and 8.5 (5.5) in group B; and during consolidation 4.7 (3.4) in group A and 4.6 (3.8) in group B (p = n.s.). Excluding the cases with consumption factors from the analysis, group B patients required 34% fewer transfusions during induction and 15.5% fewer during consolidation (p = 0.04). There were no differences between groups regarding major bleeding episodes. INTERPRETATION AND CONCLUSIONS: Our data show that the threshold for prophylactic platelet transfusion can be safely set at 10,000 microL during induction and consolidation chemotherapy for adult patients with de novo AML. PMID- 9864921 TI - Controlled-rate versus uncontrolled-rate cryopreservation of peripheral blood progenitor cells: a prospective multicenter study. Group for Cryobiology and Biology of Bone Marrow Transplantation (CBTMO), Spain. AB - BACKGROUND AND OBJECTIVE: Cryopreservation of hemopoietic progenitors for transplantation has been traditionally performed by the use of a controlled-rate freezer. Several groups have reported successful cryopreservation of progenitor cells at -80 degrees C without a controlled-rate freezer. In an attempt to elucidate whether both methods are equally efficient, we compared controlled-rate versus uncontrolled cryopreservation of peripheral blood progenitor cells (PBPC) in a prospective, multicenter study. DESIGN AND METHODS: Apheresis products from patients undergoing PBPC mobilization were split into two aliquots, and cryopreserved simultaneously by both methods, in autologous plasma plus 10% dimethylsulfoxide. Controlled-rate samples were placed into a programmable freezer with a cooling rate of 1-2 degrees C/min. Uncontrolled-rate samples were directly introduced into a -80 degrees C mechanical freezer. After thawing, cell counts, assays for viability, clonogenic cultures and CD34+ cell enumeration were performed. RESULTS: A total of 105 cases were included. No significant differences were found in viability (mean 88.8 +/- 13% in the controlled-rate group vs. 89.7 +/- 12% in the uncontrolled-rate group), nucleated cell loss (23.5 +/- 23% vs. 23 +/- 22%), mononuclear cell loss (19 +/- 23% vs. 19.1 +/- 22%), and loss of CD34+ cells (34.3 +/- 33% vs. 28.6 +/- 34%). On the other hand, recovery of granulomonocytic colony-forming units (CFU-GM), was significantly better with the controlled-rate technique, than with the non-controlled-rate method (104.3 +/ 95 vs. 86.5 +/- 80, respectively; p = 0.048). INTERPRETATION AND CONCLUSIONS: Our results indicate that both techniques are suitable for cryopreservation of PBPC, although a better recovery of committed progenitors is achieved by the controlled-rate method. Therefore, the use of controlled-rate freezer should probably be recommended. PMID- 9864922 TI - The frequency of 844ins68 mutation in the cystathionine beta-synthase gene is not increased in patients with venous thrombosis. AB - BACKGROUND AND OBJECTIVES: A frequent mutation in the cystathionine beta-synthase (CBS) gene (844ins68, a 68-bp insertion in the coding region of exon 8) was recently discovered. In the present study we investigated this mutation as a candidate risk factor for venous thrombosis. DESIGN AND METHODS: The prevalence of the 844ins68 CBS mutation was determined in 101 patients with objectively diagnosed deep venous thrombosis and in 101 healthy controls matched for age, sex and race. PCR amplification of a DNA fragment containing exon 8 of the CBS gene was employed to determine the genotypes. Additionally, Bsrl restriction enzyme digestion of the PCR products was performed in all samples from carriers of the insertion, to test for concurrent presence of a second mutation (T833C) in the CBS gene. RESULTS: The insertion was found in 21 out of 101 patients (20.8%; allele frequency 0.109) and in 20 out of 101 controls (19.8%; allele frequency 0.114), yielding a relative risk for venous thrombosis related to the 844ins68 CBS mutation close to 1.0. In addition, the T833C CBS mutation was detected in all alleles carrying the 844ins68 CBS insertion, confirming the co-inheritance of the two mutations. INTERPRETATION AND CONCLUSIONS: Our findings do not support the hypothesis that the 844ins68 mutation in the CBS gene is a genetic risk factor for venous thrombosis. PMID- 9864923 TI - von Willebrand factor contained in factor VIII concentrates of different purities supports platelet adhesion in blood samples from a heterogeneous group of patients with von Willebrand disease. AB - BACKGROUND AND OBJECTIVE: Plasma derived FVIII-VWF concentrates in which the VWF structure is reasonably maintained are recommended as substitutive therapy in VWD. Our aim was to assess platelet deposition and binding to subendothelial structures of VWF present in FVIII concentrates. DESIGN AND METHODS: Cryoprecipitate (CRY), intermediate-purity (IPC), or high-purity (HPC) FVIII concentrates were added in vitro to citrated blood samples from 11 patients affected by different subtypes of VWD, with the aim of normalizing VWF levels. Measurements of VWF:Ag, ristocetin cofactor (RiCof) activities, FVIII coagulant activity (FVIII:C), and platelet interaction with subendothelium under flow conditions (Baumgartner's perfusion method, computer-assisted morphometry, shear rate 1000 s-1, 10 min, 37 degrees C) were determined. Binding of VWF to the luminal surface of the perfused vessels was assessed by immunofluorescence microscopy. Paired t-test statistics were performed. RESULTS: Addition of FVIII VWF preparations raised VWF:Ag from baseline (BSL) values of 0.3 (SD 0.2) to averages of 1.4 (SD 0.5, p < 0.001), 1.2 (SD 0.6, p < 0.001), and 0.4 (SD 0.3) IU mL-1 after CRY, IPC, and HPC, respectively. A positive labeling for VWF was observed by immunofluorescence in vessels perfused with blood containing any of the concentrates. Platelet adhesion of 13.2 (SD 7.6), 22.4 (SD 10.8), 24.8 (SD 7.8, p < 0.03), or 22.5 (SD 4.8)% was measured in BSL, CRY, IPC, or HPC tests, respectively. INTERPRETATION AND CONCLUSIONS: Our observations support the hypothesis above the mechanisms involved in the beneficial effects of commercial concentrates in von Willebrand disease: the VWF in these concentrates has functional capacity to bind to subendothelium and to support platelet adhesion. PMID- 9864924 TI - Therapy of acute myeloid leukemia: towards a patient-oriented, risk-adapted approach. AB - BACKGROUND AND OBJECTIVE: The successful use of differentiating treatment for patients with acute promyelocytic leukemia (APL) suggests that other acute myeloid leukemias (AML) may benefit from tailored and subtype-specific therapy. Despite the fact that new drugs specifically targeting AML genetic lesions have not yet been developed, distinct karyotypic categories have been identified which may deserve differentiated treatment. In addition, molecular assays to assess response to therapy more sensitively are now available for several AML subsets. In this review, we discuss the role of genetic characterization in the therapy of AML, and the investigative efforts which we believe are still needed for the design of tailored treatment for each and every patient with this disease. DESIGN AND METHODS: The authors have been working in this field for many years and have contributed original papers, the data of which are incorporated in this article. In addition, the material analyzed in this overview includes articles and reviews covered by the Science Citation Index and Medline as well as some more recent unpublished personal observations. RESULTS: Modern therapeutic approaches to AML tend to differentiate post-induction treatment intensity according to cytogenetically defined risk categories. Such prognostic categorization is largely unsatisfactory. In fact, following the advent of newly developed molecular assays (e.g. RT-PCR and FISH), specific and prognostically relevant lesions are frequently found in patients with an apparently normal karyotype, and these patients are, therefore, re-assigned to more appropriate prognostic categories. In addition, recent studies suggest that some patients may benefit from an increase in induction intensity; rapid genetic characterization will be needed for future differentiation of initial therapy. However, preliminary investigation of AML by integrated karyotypic/molecular analyses show that no specific abnormalities are detectable in at least half of the cases. Therefore, use of genetic criteria for prognostic stratification is currently feasible in only a proportion of patients. INTERPRETATIONS AND CONCLUSIONS: The prognostic role of genetic lesions, currently identified by karyotypic studies, needs to be validated in large series of AML patients prospectively characterized by advanced molecular/cytogenetic analyses and treated uniformly. In addition, searches for new clinically relevant genetic abnormalities, and diagnostic tools for their rapid identification are urgently needed to identify prognostic categories better. Elucidation of AML gene alterations should foster basic investigation aimed at developing new drugs targeted to the specific lesion in the individual patient. Before these more specific therapeutic agents are developed, diagnostic genetic characterization should add to other well-established prognostic factors to optimize the use of the presently available therapies. PMID- 9864925 TI - Acute generalized, widespread bleeding. Diagnosis and management. AB - BACKGROUND AND OBJECTIVE: Acute generalized, widespread bleeding is often related to disseminated intravascular coagulation (DIC), a pathologic process which complicates the clinical course of many diseases and is characterized by huge amounts of thrombin and plasmin within the circulation. The final result is the consumption of platelets, coagulation factors and inhibitors, as well as secondary hyperfibrinolysis, all leading to diffuse hemorrhage and microthromboses. This review article examines the present attitudes to the diagnosis and treatment of overt DIC in clinical practice, emphasizing the importance of an accurate differential diagnosis from some other processes characterized by acute generalized, widespread bleeding. INFORMATION SOURCES: The authors have been working in this field, both at experimental and clinical levels, contributing original papers for many years. In addition, material examined in this review includes articles published in journals covered by MedLine, recent reviews in journals with high impact factor and in relevant books on hemostasis and thrombosis. STATE OF ART AND PERSPECTIVES: DIC is an intermediary mechanism of disease which complicates the clinical course of many well-known disorders. Although the systemic hemorrhagic syndrome is the predominant clinical manifestation, massive intravascular thrombosis frequently occurs contributing to ischemia and associated organ damage, making the mortality rate of this condition high. Current concepts on the pathophysiology, laboratory diagnosis and management of DIC are presented. Complex pathophysiological interrelations make the diagnosis of the etiology of the DIC difficult in clinical practice, although simple tests are useful for identification of patients with the process. Laboratory diagnosis of DIC is mainly based on screening assays, which allow a rapid diagnosis, whereas some other highly sensitive but more complex assays are not always available to routine clinical laboratories. The management of DIC is based on the treatment of the underlying disease, supportive and replacement therapies and the control of the coagulation mechanisms. Although some advances have been achieved, management decisions are still controversial, so that therapy should be highly individualized depending on the nature of the DIC and severity of clinical symptoms. Many syndromes sharing common findings with DIC, such as primary hyperfibrinolysis or thrombotic thrombocytopenic purpura, should be excluded. Finally, new therapeutic approaches to the management of this potentially catastrophic syndrome are required. PMID- 9864926 TI - Cholestasis secondary to Hodgkin's disease: report of 2 cases of vanishing bile duct syndrome. AB - Only a small percentage of patients with Hodgkin's disease become clinically Jaundiced during their disease. This Jaundice may be secondary to biliary obstruction, hemolysis, direct hepatic infiltration by the disease, drug toxicity or viral hepatitis. Vanishing bile duct syndrome secondary to Hodgkin's disease is a rare cause of cholestasis in these patients, only 13 cases having been reported so far. The authors describe 2 patients who developed severe Jaundice secondary to Hodgkin's disease due to vanishing bile duct syndrome affecting small intrahepatic bile ducts. PMID- 9864927 TI - Ultrasound image of massive inferior vena cava thrombosis causing asymptomatic subclinical disseminated intravascular coagulation. PMID- 9864928 TI - Pure red cell aplasia in autoimmune polyglandular syndrome with T lymphocytosis. AB - We report the onset of pure red cell aplasia (PRCA) in a patient with a history of polyglandular syndrome including Addison's disease, malabsorption syndrome, diabetes type I and gastric hyperplastic polyposis. An autoimmune origin for this complex disorder was not supported by the presence of organ specific antibodies, but T cells were found to be of polyclonal origin, as demonstrated by molecular analysis of T cell receptor (TCR) gene rearrangement. The pathophysiology of this case, based on laboratory findings and response to therapy, is discussed. PMID- 9864930 TI - In vitro modulation of bcl-2 protein expression, drug-induced apoptosis and cytotoxicity by interleukin-10 in chronic lymphocytic leukemia. AB - Interleukin-10 failed to modify either the percentage of bcl-2+ cells or the number of bcl-2 molecules, or to reduce 2-chlorodeoxyadenosine- and fludarabine induced apoptosis. The cytokine at 0.1 ng/mL induced an increase of cell survival both in the absence or in the presence of 2-chlorodeoxyadenosine, while no difference was appreciated with fludarabine. PMID- 9864929 TI - Fatal myelofibrosis following fludarabine administration in a patient with indolent lymphoma. AB - We report a case of fulminant myelofibrosis after administration of fludarabine in a patient diagnosed as having refractory low-grade lymphoma, progressing fatally. Myelofibrosis in the setting of an indolent lymphoma is very rare; this fact, and the short period between drug administration and fibrosis suggest an etiopathogenic link, although this potential and severe adverse effect of fludarabine has not been previously reported in the literature. PMID- 9864931 TI - Blastogenic response of activated human umbilical cord blood T-lymphocytes. AB - Donor T-lymphocytes are thought to play a crucial role in both acute and chronic graft-versus-host disease (GvHD), pathological conditions that frequently complicate allogeneic bone marrow transplantation. These diseases are described as occurring with a lower incidence and lesser severity when human umbilical cord blood (HUCB) cells, which have recently emerged as a potential source of hematopoietic progenitors, are used for transplantation. This condition is probably related to the immaturity of neonatal HUCB T cells. Lymphocyte blastogenic response to phytohemagglutinin (PHA), evaluated by means of flow cytometry, is a useful tool for testing the functional ability of T-cells to display an immune response against allo-antigens, reproducing in vitro the in vivo mechanism of activation. This study was designed to verify whether an impairment in HUCB T-cell ability to undergo an in vitro blastogenic response to mitogens could explain their reduced in vivo ability to induce GvHD. PMID- 9864932 TI - A patient homozygous for mutation 20210A in the prothrombin gene with venous thrombosis and transient ischemic attacks of thrombotic origin. AB - It is well established that genetic disorders interact with environmental factors to cause thrombotic diseases. Therefore, antithrombin, protein C, protein S deficiencies and the more recently described factor V Leiden and prothrombin mutations are currently been investigated to explain some thrombophilic states. We report the case of a 63-year-old man who developed two transient ischemic attacks and two years later an extensive femoro-iliac venous thrombosis. He was genotyped as FV R506Q negative and FII G20210A positive in homozygous state (FII 20210AA). PMID- 9864934 TI - Circulating thrombopoietin and interleukin-6 in newly diagnosed autoimmune versus aplastic thrombocytopenia. AB - Circulating thrombopoietin and interleukin-6 concentrations were investigated in two different settings of thrombocytopenia. Twenty patients with autoimmune thrombocytopenic purpura (ATP), 12 patients with aplastic anemia (AA) and 15 healthy subjects were studied. Thrombopoietin was significantly increased in AA and deficient in ATP. Interleukin-6 was significantly increased in ATP, compared to both other groups. PMID- 9864933 TI - Role of autologous bone marrow transplantation as consolidation therapy in acute promyelocytic leukemia patients in complete remission. AB - Autologous bone marrow transplantation (ABMT), which consents a low mortality rate, has been proposed as an alternative approach to maintenance chemotherapy in patients with acute promyelocytic leukemia (APL) in first complete remission irrespective of the patients' molecular status. Sixteen patients with acute APL in complete remission were submitted to ABMT and were analyzed for the presence of the PML-RARa fusion gene by reverse transcription-polymerase chain reaction (RT-PCR). Our study demonstrated that continued positivity of PCR analysis before ABMT could predict subsequent relapse in patients who undergo un-purged ABMT procedures. PMID- 9864935 TI - Tetanus: still "inexcusable". PMID- 9864936 TI - A laparoscopic update. PMID- 9864937 TI - Laparoscopic ultrasound: a valuable adjunct to laparoscopic surgery. AB - Laparoscopic ultrasound represents a recent merger in the laparoscopic technology and intraoperative ultrasound and shows a diagnostic accuracy higher than preoperative studies. Laparoscopic ultrasound can be used during laparoscopic cholecystectomy to screen the bile duct. It is particularly useful for diagnosing and staging malignancies, including hepatobiliary, pancreatic and gastroesophageal cancers. By demonstrating the interior of organs and deep structures, it can compensate for the limitation of laparoscopic examination. Laparoscopic ultrasound will become a valuable adjunct to laparoscopic surgery. PMID- 9864938 TI - Laparoscopic inguinal herniorrhaphy: the new gold standard of hernia repair? AB - The surgical treatment of the common inguinal hernia has been one of the most analyzed and debated topics in medicine. Recently, with the success of laparoscopic cholecystectomy, interest in minimally invasive surgical techniques has led to it's application for inguinal hernia repair. Current laparoscopic herniorrhaphies are based on the principles of conventional open preperitoneal repairs and are classified into two types: 1) transabdominal preperitoneal repair (TAPP) and 2) totally extraperitoneal repair (TEP). Common advantages to both techniques include a decrease in postoperative pain, earlier return to normal activity, and improved cosmesis. Both laparoscopic techniques have the disadvantage of requiring general or regional anesthesia and increased procedural costs. Lastly, there is a concern that laparoscopic hernia repair has not been around long enough to know the risk of late recurrences. Laparoscopic herniorrhaphy, however, is a viable alternative to standard open inguinal hernia repair. PMID- 9864939 TI - Laparoscopic staging of malignant disease. AB - The earliest applications of laparoscopy were for diagnostic procedures. The goal was to make a diagnosis while sparing the patient a major, and often futile, operation. However, the view was limited and it was not possible to palpate organs or masses. The recent development of advanced laparoscopic techniques and ultrasound have improved our view and restored our "sense of touch". These innovations bring the goal of minimally invasive diagnosis and staging closer to reality. This paper reviews the current literature on the laparoscopic staging of cancer with an emphasis on patient selection, diagnostic accuracy, and the reduction in morbidity which can be achieved. PMID- 9864940 TI - Advanced laparoscopy: "the next generation". The adrenal, kidney, spleen, pancreas, and liver. AB - Following the rapid acceptance of laparoscopic cholecystectomy, a few adventurous surgeons "dared to boldly go where no one had gone before." They sought to achieve the same reduction in morbidity while accomplishing the same surgical goals. This paper will briefly review the current status of minimal access surgery for the solid organs of the abdomen. It will focus on the indications, risks, limitations, and on the balance between the trauma of access and the trauma of the procedure itself. As new techniques and equipment emerge and experience and data are accumulated, this balance may shift. Some of these procedures are in their infancy while others are rapidly becoming the new "Gold Standard". PMID- 9864941 TI - Is Wiskott--Aldrich syndrome a cell trafficking disorder? PMID- 9864942 TI - HLA and cancer: from research to clinical impact. PMID- 9864943 TI - HIV-I Tat: a polypeptide for all seasons. PMID- 9864944 TI - The Roman god Janus: a paradigm for the function of CD43. PMID- 9864945 TI - Non-traditionally derived CTL epitopes: exceptions that prove the rules? PMID- 9864946 TI - Nitric oxide in autoimmune disease: cytotoxic or regulatory mediator? PMID- 9864947 TI - Catecholaminergic suppression of immunocompetent cells. PMID- 9864948 TI - Chemokines and chemokine receptors in T-cell priming and Th1/Th2-mediated responses. PMID- 9864949 TI - The organization of mature T-cell pools. PMID- 9864950 TI - Why are there two rat TAPs? PMID- 9864951 TI - A common epitope on gp41, IFN-alpha and IFN-beta induces protective activity. PMID- 9864952 TI - Peripheral T-cell self-reactivity and immunological memory. PMID- 9864953 TI - On the complete physician. PMID- 9864954 TI - Leaders in medicine. M. Joe Crosthwait. PMID- 9864955 TI - Diabetes mellitus and major depression: considerations for treatment of Native Americans. AB - Non-insulin dependent diabetes mellitus (NIDDM) extracts a heavy toll on the Native American community in the United States. Evidence indicates that patients with NIDDM are three times more likely to have a co-existing diagnosis of depression. Untreated major depression unfavorably impacts the complication rates of NIDDM. Thus, Native Americans who are at increased risk for NIDDM are likely to be at increased risk for major depression. Physicians in Oklahoma should be aware of important treatment issues when selecting an antidepressant medication to treat major depression in Native Americans with NIDDM. Treatment options for major depression in the context of diabetes are discussed. Evidence currently indicates that the serotonin reuptake inhibitors (SSRIs) have significant advantages and a more favorable side effect profile for the treatment of depression in patients with diabetes mellitus. PMID- 9864956 TI - 47-year-old man with left flank pain: a clinical pathological correlation conference of the University of Oklahoma College of Medicine. PMID- 9864957 TI - Physician-older patient communication during the transition from independence to dependence. AB - This article reviews recent research regarding the communication between physicians and their older patients who are passing from independence to dependence. Major findings from these studies are summarized with a discussion of how the context of the medical encounter, the physical, cognitive and language changes related to age, the amount of time permitted for a medical encounter, the managed care environment, and the presence of a companion can affect competent communication between a physician and an older adult. The article concludes with a comment about the need to educate current and future physicians on the complexity of communicating effectively with older patients. PMID- 9864958 TI - Comparison of influenza immunization rates for Oklahoma Medicare patients: 1995, 1996 and 1997. AB - The Health Care Financing Administration has reported influenza immunization rates since 1994. The Department of Health and Human Services has set a minimum national target rate for the annual immunization of the elderly population at 60 percent, as published in Healthy People 2000. The Oklahoma Foundation for Medical Quality analyzed the Medicare claims data for Oklahoma for the 1995, 1996, and 1997 influenza seasons. Additionally, we reviewed the Behavioral Risk Factor Surveillance System influenza immunization data for 1995. Claims data for the 1997 influenza season show the immunization rate for the Medicare population of Oklahoma is 41.4 percent. The immunization rate for the African-American Medicare population was 22.3 percent for 1997, compared with 42.2 percent for the Caucasian population. The ten most populous counties in the state had a 9-percent higher rate of immunization than the other 67 counties. The Medicare population in Oklahoma is not receiving the influenza vaccination at the target rate. Especially underserved are the African-American and non-urban populations. There appear to be opportunities for improvement in the provision of the influenza vaccination for the Medicare population of Oklahoma. PMID- 9864959 TI - PLICO response. PMID- 9864960 TI - The role of vitamin E in T-cell differentiation and the decrease of cellular immunity with aging. AB - Spontaneously hypertensive rats (SHR) as a model for aging were used in this experiment and fed a regular (50 IU/Kg diet) or high vitamin E (500 IU/Kg diet) diet for 6 weeks. At 12 weeks old, they were killed and assayed. Although proliferation of thymic lymphocytes was significantly decreased in SHR fed the regular diet compared to Wistar Kyoto rats (WKY) fed the same diet, high vitamin E diet enhanced proliferation of thymic lymphocytes in SHR to almost the levels in WKY fed the regular diet. In addition, the expressions of both CD4 and CD8 antigens on CD+CD8+ T cells, immature T cells existing in thymic cortex, were also decreased in SHR, and significantly improved by high vitamin E diet. These results suggest that high vitamin E diet enhances thymic lymphocyte proliferation through increased T-cell differentiation in thymus. Then, the effect of vitamin E on T-cell differentiation in thymus was investigated by using male Fisher rats. Rats were divided into three groups; vitamin E-free, regular and high vitamin E groups and fed a diet containing various levels of vitamin E (0, 50 and 500 IU/Kg diet) for 7 weeks. Although the percentages of CD4+CD8- and CD4-CD8+ T cells in thymocytes were significantly greater in the high vitamin E group, the percentage of CD4+CD8- T cells inversely decreased in the vitamin E-free group compared to the regular group. We have tried to investigate the mechanism of the increased T cell differentiation in thymus of rats fed the high vitamin E diet through cytokine production, and thymic epithelial cell (TEC) and macrophage functions. We have found that vitamin E enhances T-cell differentiation through the increase of not macrophage but TEC function in thymus, which is associated with the increased binding capacity of TEC to immature T cells via increased expression of adhesion molecule, ICAM-1. These results suggest that vitamin E is a potent nutrient for promoting health in the aged via the improvement of cellular immunity decreased with aging. PMID- 9864961 TI - State of the art: "diastology" research 1998. AB - The performance of the left ventricle (LV) during diastole is defined by the pressure-volume relationship. Consequently, noninvasive techniques have been limited in the evaluation of diastolic function by their inability to evaluate intracardiac pressure, particularly LV filling pressure and end-diastolic pressure. Abnormalities of LV diastolic function play a major role in producing the clinical signs and symptoms of heart failure. Previous studies have demonstrated that the transmitral flow (TMF), pulmonary venous flow (PVF) and left atrial appendage flow (LAAF) velocity patterns determined by pulsed Doppler echocardiography are useful parameters for evaluating left atrial (LA) and LV hemodynamic events. However, these variables are influenced by loading conditions, particularly preload. Furthermore, it has become increasingly clear that abnormalities of LV diastolic function, such as relaxation and filling, can be assessed precisely using color Doppler M-mode echocardiography and pulsed tissue Doppler imaging irrespective of preload. This review presents a clinical approach to understanding the hemodynamic abnormalities of the LA and LV in a variety of cardiac diseases using these new modalities. In addition, the limitations of these techniques are discussed. PMID- 9864962 TI - A hydrolase enzyme inactivating endogenous ligands for cannabinoid receptors. AB - Cannabinoids are psychoactive components of marijuana, and bind to specific G protein-coupled receptors in the brain and other mammalian tissues. Anandamide (arachidonoylethanolamide) was discovered as an endogenous agonist for the cannabinoid receptors. Hydrolysis of anandamide to arachidonic acid and ethanolamine results in the loss of its biological activities. The enzyme responsible for this hydrolysis was solubilized, partially purified from the microsomes of porcine brain, and referred to as anandamide amidohydrolase. In addition to the anandamide hydrolysis, the enzyme preparation catalyzed anandamide synthesis by the condensation of arachidonic acid with ethanolamine. Several lines of enzymological evidence suggested that a single enzyme catalyzes both the hydrolysis and synthesis of anandamide. This reversibility was confirmed by the use of a recombinant enzyme of rat liver overexpressed in COS-7 cells. However, in consideration of the high Km value for ethanolamine as a substrate for the anandamide synthesis, the enzyme was presumed to act as a hydrolase rather than a synthase under physiological conditions. The recombinant enzyme acted not only as an amidase hydrolyzing anandamide and other fatty acid amides but also as an esterase hydrolyzing methyl ester of arachidonic acid. 2 Arachidonoylglycerol, which was found recently to be another endogenous ligand, was also efficiently hydrolyzed by the esterase activity of the same enzyme. The anandamide hydrolase and synthase activities were detected in a variety of rat organs, and liver showed by far the highest activities. A high anandamide hydrolase activity was also detected in small intestine but only after the homogenate was precipitated with acetone to remove endogenous lipids inhibiting the enzyme activity. The distribution of mRNA of the enzyme was in agreement with that of the enzyme activity. PMID- 9864963 TI - Role of virus-induced apoptosis in a host defense mechanism against virus infection. AB - Many animal viruses are known to induce apoptosis in infected cells. This virus induced apoptosis has been often described as a mechanism of host defense against virus infection, based on the finding that mutants of an insect virus with the ability to induce extensive apoptosis in some cells cannot grow in the same cells. In animal virus infection, we have shown that (1) viruses can somehow overcome this defense mechanism and that (2) virus multiplication in the apoptotic cells is not as completely suppressed as in the insect virus infection. These results suggest that, in the case of animal viruses, the virus-induced apoptosis does not play the same role in the host defense system as in insect cells. However, by examining the virus infection under the conditions comparable to the infection in vivo, we demonstrated the defensive role of apoptosis in animal virus infection. PMID- 9864964 TI - Effects of electromagnetic fields on membrane ion transport of cultured cells. AB - We have studied the mechanisms of ion transport mediated by Na+/K(+)-pump and Na+,K+,Cl(-)-cotransport pathway of HeLa cells using Rb+ as an analog for K+, and proposed models of binding of ions for the transport pathways. Also, we clarified the relation between ion and water movements in the cells. Based on these findings, we have studied the effects of homogeneous and time-varying magnetic fields on the ion transport activity. The research presented here covers (i) brief explanations of our kinetic studies on the ion transport pathways for promoting understanding of the effects of magnetic fields on the pathways, (ii) our and other reports of the effects of magnetic fields on ion transport systems. PMID- 9864965 TI - Pathophysiology and treatment of cerebral ischemia. AB - This article describes the pathophysiology of, and treatment strategy for, cerebral ischemia. It is useful to think of an ischemic lesion as a densely ischemic core surrounded by better perfused "penumbra" tissue that is silent electrically but remains viable. Reperfusion plays an important role in the pathophysiology of cerebral ischemia. Magnetic resonance imaging (MRI) and histological studies in rat focal ischemia models using transient middle cerebral artery (MCA) occlusion indicate that reperfusion after an ischemic episode of 2- to 3-hour duration does not result in reduction of the size of the infarct. Brief occlusion of the MCA produces a characteristic, cell-type specific injury in the striatum where medium-sized spinous projection neurons are selectively lost; this injury is accompanied by gliosis. Transient forebrain ischemia leads to delayed death of the CA1 neurons in the hippocampus. Immunohistochemical and biochemical investigations of Ca2+/calmodulin-dependent protein kinase II(CaM kinase II) and protein phosphatase (calcineurin) after transient forebrain ischemia demonstrated that the activity of CaM kinase II was decreased in the CA1 region of the hippocampus early (6-12 hours) after ischemia. However, calcineurin was preserved in the CA1 region until 1.5 days after the ischemic insult and then lost; a subsequent increase in the morphological degeneration of neurons was observed. We hypothesized that an imbalance of Ca2+/calmodulin dependent protein phosphorylation-dephosphorylation may be involved in delayed neuronal death after ischemia. In the treatment of acute ischemic stroke, immediate recanalization of the occluded artery, using systemic or local thrombolysis, is optimal for restoring the blood flow and rescuing the ischemic brain from complete infarction. However, the window of therapeutic effectiveness is very narrow. The development of effective neuroprotection methods and the establishment of reliable imaging modalities for an early and accurate diagnosis of the extent and degree of the ischemia are imperative. PMID- 9864966 TI - Effects of interferon-alpha and gamma on development of LAK activity from mononuclear cells in breast cancer patients. AB - We examined the effect of recombinant IFN-alpha and IFN-gamma on induction of LAK cells from peripheral blood mononuclear cells (PBMNCs) in 7 pre-operative breast cancer patients and 4 healthy volunteers. Significant LAK activity was developed from PBMNCs of pre-operative breast cancer patients and healthy volunteers after incubation for 4 days with IL-2 (presence of IL-2 vs. absence of IL-2). Incubation of PBMNCs of pre-operative breast cancer patients with 1000 U/ml of IFN-alpha for 4 days suppressed the LAK activity significantly (P < 0.05). By contrast, incubation of PBMNCs of pre-operative patients with 1000 U/ml of IFN gamma for 4 days increased the LAK activity significantly (P < 0.05). Significant cytotoxicity against MCF-7 cells (estrogen receptor positive human breast cancer cell line) was developed from PBMNCs of pre-operative breast cancer patients at 20:1 and 40:1 E/T ratios after incubation for 4 days with IL-2 (absence of IL-2 vs. 20:1 or 40:1, P < 0.05, P < 0.05), whereas PBMNCs of healthy volunteers did not. Stimulation of LAK cells with IFN-gamma produced a significant augmentation of cytotoxic activity against MCF-7 (P < 0.05), while IFN-alpha suppressed the cytotoxicity significantly (P < 0.05). These findings suggested that combined stimulation by IFN-gamma and IL-2 might be a reasonable treatment for breast cancer patients. PMID- 9864967 TI - Fibrinogenolytic activity of a novel trypsin-like enzyme found in human airway. AB - Previously we isolated a new trypsin-like enzyme designated human airway trypsin like protease (HAT) from human sputum. In this study, we examined in vitro whether HAT was related to the prevention of fibrin deposition in the airway lumen by cleaving fibrinogen. In mucoid sputum samples from patients with chronic airway diseases, the concentration of fibrinogen, as measured by ELISA, was in the range of 2-20 micrograms/ml, and trypsin-like activity, as measured by spectrofluorometry was in the range of 10-50 milliunits (mU)/ml. We showed by gel filtration that the trypsin-like activity of mucoid sputum was mainly due to HAT. We examined the effects of HAT on human fibrinogen at pH 7.4 and 8.6. Fibrinogen was used at concentrations of 4-2,000 micrograms/ml and HAT purified from sputum at concentrations of 0.6-10 mU/ml. As shown by SDS-polyacrylamide gel electrophoresis, HAT cleaved fibrinogen, especially its alpha-chain, regardless of the concentration of fibrinogen. Pretreatment of fibrinogen with HAT resulted in a decrease or complete loss of its thrombin-induced clotting capacity, depending on the duration of pretreatment with HAT and the concentration of HAT. From these results we postulated that HAT may participate in the anticoagulation process within the airway, especially at the level of the mucous membrane, by cleaving fibrinogen transported from the blood stream. PMID- 9864968 TI - Contrast medium-removing effect of hemofiltration and hemodiafiltration. AB - The contrast medium-removing effect of hemofiltration (HF) and hemodiafiltration (HDF) was experimentally investigated using a bovine blood tank model. HF and HDF were performed at a blood flow rate of 100 ml/min with a polysulfone hemofilter (PS filter-CF; membrane area: 0.7 m2). Two hundred milliliters of iomeprol (300 mgI/ml) was administered by a single injection into 4 liters of bovine blood. The blood half-lives of iomeprol were 1.0 hr for the high flow rate HDF group [replacement fluid flow rate (QF): 10 ml/min and dialysate flow rate (QD): 40 ml/min], 1.8 hr for the HDF group (QF: 10 ml/min and QD: 10 ml/min), and 3.8 hr for the HF group (QF: 10 ml/min). The mean clearance rates were 39.7 ml/min for the high flow rate HDF group, 21.4 ml/min for the HDF group, and 12.0 ml/min for the HF group. Iomeprol was mostly excreted in the waste fluid. It is concluded that HDF can remove contrast media more effectively than HF. PMID- 9864969 TI - Prognostic significance of Ki-67 and p53 antigen expression in carcinomas of bile duct and gallbladder. AB - Ki-67 and p53 protein expression was evaluated immunohistochemically in 32 patients with intrahepatic, extrahepatic bile duct and gallbladder carcinomas, who underwent surgery at First Department of Surgery, The University of Tokushima School of Medicine. p53 expression was found more in the well differentiated group than poorly differentiated group (p = 0.007). MIB1 labelling index (MIB1 LI) was higher in EHC than in GBC (p = 0.0061). MIB1 LI (T), (MIB1 LI in tumor) was higher in cases with lymph node metastasis than in those without lymph node metastasis (p = 0.0189). Moreover, MIB1 LI (L) (MIB1 LI in metastasized lymph node) was higher in poorly differentiated than in well differentiated carcinoma (p = 0.0404). Prognostically, patients with high MIB1 LI (T) (> 56.93) had a worse prognosis after surgery than those with low MIB1 LI (T) (p < 0.05). There was no association between p53 positive tumors and MIB1 expression. These results suggest that cancer cell proliferative activity was markedly increased in cases with EHC compared to those with GBC and the poorly differentiated and lymph node metastasis group. MIB1 LI in tumor was found to be a good prognostic indicator whereas there was no association of p53 positive tumor with MIB1 expression and prognosis of the patients. PMID- 9864970 TI - Effect of partial pancreatectomy on beta-cell mass in the remnant pancreas of Wistar fatty rats. AB - Wistar fatty rat, which has been established by transferring the fa gene of Zucker fatty rat to the Wistar Kyoto rat, has many features in common with human NIDDM. It exhibits hyperglycemic obesity with hyperinsulinemia and insulin resistance. It is unclear, however, whether a defect in the beta-cell proliferation is related to the onset of diabetes mellitus together with insulin resistance in this model rat. To determine this, we compared non-fasting plasma glucose levels, insulin content and beta-cell mass in the remnant pancreas of Wistar fatty rats with those in their diabetic-resistant lean counterparts after a 70% partial pancreatectomy. We also examined whether such a defect, if present, could be improved by either phlorizin or nicotinamide. We further investigated if there were any differences in these parameters between the phenotypically identical but genotypically different Wistar lean rats with a gene type of homogeneous Fa/Fa and that of heterogeneous Fa/fa. Male rats, 6 weeks of age, were allocated at random into two groups: 70% pancreatectomy (Px) and sham pancreatectomy (sham). A sustained hyperglycemia was evident in the Px Wistar fatty rats after surgery, which was accompanied by a reduction of insulin content and beta-cell mass in the remnant pancreas. The changes in insulin content and beta-cell mass were unaffected by restoration of normoglycemia, induced by phlorizin injection. The administration of nicotinamide partially ameliorated the sustained hyperglycemia by a slight but not significant increase in beta-cell mass. No discernible difference in the above parameters was observed between the Wistar lean rats with Fa/Fa and those with Fa/fa. These findings suggest that Wistar fatty rats have a poor capacity for proliferation of pancreatic beta cells, which causes the onset of overt diabetes along with insulin resistance due to extreme obesity. PMID- 9864971 TI - Rapid and convenient method of autoradiography for DNA cloning using digital imaging analysis. AB - Digital image analysis has been used for various biochemical and molecular biological analyses instead of autoradiography with X-ray film. However, in such cases the data manipulated by an imaging analyzer was generally printed out on normal printing paper. With normal paper, it is difficult to align the signal or its position relative to the original sample. Here in, we demonstrate it to be convenient and accurate to align signal obtained by imaging analyzer with OHP film. PMID- 9864972 TI - Effect of administration of ursodeoxycholic acid at bedtime on cholesterol saturation of hepatic bile in Japanese patients with gallstone. AB - The administration of a single, daily 600 mg dose of ursodeoxycholic acid (UDCA) at bedtime and 3-200 mg doses per day at mealtime was conducted for 6 patients with gallstone and choledocholithiasis who were undergoing biliary drainage for the purpose of improving jaundice. Hepatic bile was collected from a drainage tube after a lapse of time in order to compare the bile acid compositions and cholesterol saturation index (SI) in bile for the 2 protocols. A significant increase in UDCA levels in hepatic bile was observed after both UDCA administration at bedtime and mealtime, but the effect of bedtime administration was significantly greater than that of mealtime administration. Whereas levels of cholic acid and chenodeoxycholic acid (CDCA) decreased for the case of bedtime administration, this was not detected for mealtime administration, although no significant differences among the mean interval values were observed. A significant in difference decreased SI was observed during UDCA bedtime administration, but not during mealtime administration, compared to the SI before administration. This suggests a decreased cholesterol excretion into the bile. Based on these findings and from the point of view of compliance, bedtime administration of UDCA appears to be an effective method. PMID- 9864973 TI - Description and valuation of health-related quality of life among the general public in Japan by the EuroQol. AB - OBJECTIVE: Health-related quality of life plays an important role in assessing the effectiveness of health care. The EuroQol is a generic instrument for describing and valuing health-related quality of life. To elicit health state descriptions and their preferences among the general public in Japan and compare them with cross-national data, a feasibility study for applying the EuroQol to the general public in Japan was carried out. SUBJECTS AND METHODS: The subjects were 120 people aged 40s-60s randomly selected in a suburban area at Aichi Prefecture in Japan. In assessing health states and their preferences, the EuroQol valuation instrument (version 12, 1991) translated into Japanese was used. The questionnaires were distributed and collected by public health nurses. The valid responses (rate) were 89 (74%). RESULTS: The mean scores (raw scores) using the visual analogue scale (VAS) for one's own health was 89.2. No statistically significant difference in VAS scores was observed for both sex and age. The contribution ratio of own health status, sex and age was 0.326 (p < 0.0001). The main independent variables were three dimensions of health status. Valuations for core health states varied from 96.3 (no problem in each health status) to 6.8 (dead). These VAS scores in Japan were correlated with those from other countries (p < 0.001). A multivariate analysis indicated that bias from own health status on preference valuations for core health states was not observed. CONCLUSION: The health states and their preferences among the general public in Japan were estimated by using the EuroQol. The results show the feasibility of evaluation for health states quantitatively. Moreover, this study suggested cross national and cross-cultural applicability of the EuroQol. PMID- 9864974 TI - A case of sarcoidosis associated with chronic eosinophilic pneumonia. AB - A 38-year-old man was hospitalized in our university hospital because of pulmonary opacities with bilateral hilar and mediastinal lymphadenopathy seen on chest radiograph. Eosinophilia was observed in the circulation and bronchoalveolar lavage (BAL) fluid. Histological examination revealed noncaseating epithelioid granulomas and eosinophilic infiltration in the lung. Based on these findings, a diagnosis of sarcoidosis combined with chronic eosinophilic pneumonia was made. The infiltrates on chest radiograph and BAL eosinophilia were promptly reduced with corticosteroid therapy, but only mild reduction was observed in diffuse nodular shadows and hilar and mediastinal lymphadenopathy, and high amounts of lymphocytes in BAL fluid remained. Increased IFN-gamma, IL-4 and IL-5 were detected in the BAL fluid, and corticosteroid therapy reduced IL-4 and IL-5 (Th-2 cytokines) but not IFN-gamma (Th-1 cytokine). These cytokine levels in BAL fluid were intimately correlated with the clinical course of sarcoidosis and chronic eosinophilic pneumonia. PMID- 9864976 TI - [Surgical treatment of juxtarenal occlusion of the abdominal aorta]. AB - Incidence, etiology and clinical course of chronic juxtarenal occlusion of the abdominal aorta have been studied. It was established that this type of lesion among other forms of occlusive disease of the aorto-ileal segment is encountered in 8.1% of patients. Inflammatory diseases of arteries prevailed as an etiologic factor. Diagnosis of this disease should be based on the use of complex up-to date methods of examination. Roentgeno-contrast aortography in lateral position is of paramount importance in detection of upper level of thrombus spreading in the aorta. Original method of operative procedure used in 20 patients, has been developed with due regards for location of the occlusion, character of lesion of aortic walls and prophylaxis of intraoperative embolic complications. Restoration of blood flow in lower extremities without any intraoperative complications was obtained in all operated patients. In all suffering from symptomatic hypertension normalization of arterial pressure was obtained after the operation. Postoperative mortality rate made up 4.4%. The results obtained confirm high effectiveness of the above surgical treatment of patients with juxtarenal occlusion of the abdominal aorta. PMID- 9864975 TI - [Surgical tactics in the treatment of postoperative thrombosis of reconstructed arteries]. AB - The trial included 894 patients in postoperative period with thrombosis of reconstructed arteries, who had undergone since 1975 1100 repeated reconstructive operations. Vast experience of reoperations on the vessels allowed to summarize various types of reoperations, to systematize them and to propose new classification. Surgical policy was determined by the causes of acute thrombosis and the time of its duration. Evaluation of the results of repeated reconstructions has revealed, that patency of the by-passes in the aorto-femoro popliteal area in long-term postoperative period is significantly lower than after surgery on renal arteries and brachiocephalic branches. A significant difference in patency in femoro-popliteal and tibial segments is revealed since the 2nd year, in aorto-femoral-since the 4th year after surgery. The function of the shunts in patients with nonspecific aortic arteritis in follow up period is significantly lower than in patients with atherosclerosis beginning from the 2nd year of follow-up. The repeated reconstructive operations for early and late thromboses in femoro-tibial segment are almost ineffective. Up to year 3 after the repeated operation patients with stage II of ischemia after Fontaine make up 19%, and up to year 5--only 9%. In the rest of the patients chronic critical ischemia with the loss of the limb is develops. PMID- 9864977 TI - [Catheterization translumbar angiography of the aorta and its branches]. AB - Method of complex angiographic examination and treatment from translumbar approach (CTLA) in patients with diffuse occlusive lesions of arterial bed was developed. 529 patients (516 males and 13 females) aged from 20 to 80 years (53.1 +/- 1.6 years) were examined and operated with the use of translumbar approach: they underwent various angiographic examinations of peripheral, visceral and coronary arteries as well as interventions on arteries. The quality of contrasting of arterial bed in CTLA was higher in comparison with results of punction translumbar aortography and intravenous digital angiography, complications rate being minimal. Application of CTLA in practice made it possible to perform practically all kinds of angiography and many invasive procedures in patients with occlusions of peripheral arteries and after reconstructive operations on vessels. PMID- 9864978 TI - [Distraction osteosynthesis in occlusive diseases of the limb arteries]. AB - The method of distraction osteosynthesis was used in 265 patients with arterial occlusions of the upper and lower limbs, which developed due to obliterative thromboangiitis and atherosclerosis. Surgical treatment of patients with distal type of occlusive disease of the arteries was followed by steady compensation of ischemic disturbances after bone chip formation and its dosage across transfer with formation of distraction regenerate transforming into mature osseous tissue. Favourable results of the treatment were observed in 236 patients (88.9%). They persisted during 3 years in 86% of patients. Analysis of outcomes after treatment demonstrated the best results in patients with grade II ischemia. Distraction osteosynthesis ensured protracted remission of the disease and elimination of its symptoms. PMID- 9864979 TI - [Extirpation of the thoracic part of the esophagus using left lateral thoracoabdominal and cervical approach]. AB - From 1988 to 1994 year extirpation of thoracic part of the esophagus with the use of left lateral thoracoabdominal and cervical approach was carried out in 81 patients with cancer of middle (27) and lower (25) thirds of the esophagus and cardia (29). Tumor was removed en bloc with regional lymph nodes, dissected stomach was fixed to the esophageal stump and transferred to the neck, where esophago-gastral anastomosis was created by uninterrupted one-layer suture. 5 patients (6.2%) died after the operation, insufficiency of sutures of the anastomosis was detected in 5 (6.2%) patients, the fistula being spontaneously closed up. Mean duration of the operation made up 309 +/- 48 min, hospitalization 18.4 + 10.9 days. Cumulative actuarial survival rate made up: 1- years--58.9 +/- 5.7%, 3 years--26.9 +/- 6.3%, 5 years-- 20.8 +/- 6.2%. Mean duration of life after the operation was 27.8 +/- 3.4 months. PMID- 9864980 TI - [Video-thoracoscopic surgery]. AB - The experience in application of 24 videothoracoscopical operations in diseases and traumatic injuries of the organs of thoracic cavity (15-diagnostic and 9 curative ones) is reviewed. Histological verification of the diagnosis after the operative procedures was obtained in all 15 patients. The technique of TV thoracoscopic operations is outlined. Postoperation period in patients ran well, there were no substantial disturbances in function of external breathing, motor activity has restored on the second day after the operation. PMID- 9864981 TI - [Use of plasma scalpel and one-layer suture in resection of the stomach]. AB - 110 cases of resection of the stomach with transection of the stomach and the duodenum by plasmic scalpel and suturing of curvatura minor of the stump and anastomosis with the use of original one-layer sero-muscular-submucosal suture have been analysed. Changes in gastric wound at the site of plasma transection were evaluated microscopically. It was established that plasma transection of the stomach is aseptic, hemostatic, thermal trauma is minimal and does not exceed the level of 3-4 mm from transection line. Favourable conditions for applications of one-layer suture contribute much to a decrease of the number of stitch puncture, to rejection of catgut, to reduction of tissue mass involved and deformation of intestinal wall. In the early postoperative period wound and anastomoses healing occurred without substantial inflammatory changes. 109 patients were operated by Billroth-1 and 1 patient--by Billroth-II. One patient died (lethality 0.9%). Early postoperative complications occurred in 7.2% of operated patients. PMID- 9864982 TI - [Choice of the mode of intubation and decompression of the small intestine in acute obstruction]. AB - On the basis of case records analysis of 370 patients from 1107 with acute small bowel obstruction (ASBO) treated in hospital, the authors have classified methods of intubation and decompression of the small bowel and modes of the use of decompression probe. Classification of clinical course of ASBO is proposed for substantiation of application of various modes of intubation and decompression depending on the progression stage of ASBO and medical technology for treatment of such kind of patients. PMID- 9864983 TI - [Treatment of stage I-IIA breast cancer of medial-central localization]. AB - 702 patients with breast cancer were followed-up after, sparing and radical operations combined with radiation and polychemotherapy. Of adjuvant treatments, radiation therapy showed to be more effective than chemotherapy. Methods of operative treatment combined with radiation are not essential in the treatment of early stages of breast cancer. PMID- 9864984 TI - [Is it possible to improve the quality of life of patients who have undergone pelvic evisceration?]. AB - Locally spread cancer makes up considerable percent (20-30%) in statistical structure of rectal tumors. In cases of cancer spread into the area of urine bladder triangle the operation of choice is pelvic evisceration. From 1977 to 1997 in the State research Centre of the Ministry of Health for Coloproctology pelvic evisceration in cancer spread to back wall of urine bladder in the area of triangle was carried out in 22 patients (20 male and 2 female). Mean age was 43.4 (29-56) years, 16 patients have undergone typical infralevator pelvic evisceration. There were no intraoperative lethality. Postoperative lethality made up 6.3%, complications--68.8%, 5-years survival rate--25%. Presence of two fecal fistulas on the anterior abdominal wall has decreased considerably the quality of life of the patients. Since 1993 the conception of preservation and restoration of natural passage of urine and bowel contents was adopted. In 4 cases infralevator pelvic evisceration with various types of ileocystoplasty and pull-through of colon into small pelvis with creation of smooth muscle cuff in perineal colostomy was carried out. In 2 patients evisceration was of supralevator-character cystoplasty of local tissues and performance of coloanal anastomosis were carried out. The application of reconstructive-restorative ways in coloproctology and urology considerably contributed to the improvement of the quality of life of the patients after pelvic evisceration. PMID- 9864985 TI - [Use of two-dimensional fluorescence diagnosis in oncological practice]. PMID- 9864986 TI - [Evaluation of wound healing process]. AB - Mathematical modelling of wound healing process gave use to new nontraditional methods for treatment of festered wounds of various origin: volume-loaded activated water with pH from 2.4 to 11.3, being used for dressings and prepared in electrohydrolyzer with graphite anode (which possessed electric potential +1.200 mV, electric potential of furacillin solution 1:500 widely used for dressing, is equal to +300 mV). This approach to the treatment of festered wounds in complex with radical surgery allowed to substantially improve the results of treatment in 1979-to 1994 in comparison with the period 1971-1988: the rate of repeated hospitalizations lowered 3.9-todd, frequency of allergic complications- 5 told, mortality rate--1.5 told and without oncological and vascular diseases--5 told. The duration of hospital stay and temporary unemployment diminished 2 times. PMID- 9864987 TI - [Bile duct decompression and immune correction in the surgical treatment of mechanical jaundice of non-tumor origin]. AB - The aims of the study were: 1) to determine optimal duration of bile ducts decompression (BDD) and terms for radical operation performance; 2) to define criteria of effectiveness of the decompression and readiness of the patients for radical surgical treatment; 3) to analyze disorders in liver function and indices of immunoreactivity in relation to the method and duration of BDD; 4) to develop method of immunocompression for increase of effectiveness of BDD. The patients were divided in to three groups. 28 patients of group 1 had duration of jaundice up to 5 days and bilirubinemia up to 86.3 +/- 3.12 mmol/l. 35 patients of group 2 jaundice up to 15 days and bilirubinemia up to 184.7 +/- 4.22 mmol/l. In group 342 patients had bilirubinemia up to 124.4 +/- 2.23 mmol/l and cholangitis. External decompression of bile ducts quickly promotes restoration of drainage function of the liver and normalization of biochemical indexes. Restoration of immunological indexes in groups 2 and 3 is progressing slowly and demands immunocorrection. Indexes of immunoreactivity and biochemical data of functional status of the liver could serve as criteria of effectiveness of bile ducts decompression and possibility of radical surgical treatment. Regional immunocorrection is indicated for the patients' preparation for surgery. PMID- 9864988 TI - [Methods of lessening operational risks: juridical aspects (lecture)]. PMID- 9864990 TI - [Massive hemorrhage into the cavity of giant common bile duct cyst after transcutaneous transhepatic cholangiostomy]. PMID- 9864989 TI - [One-stage resection of 7 organs in cancer of the ascending colon]. PMID- 9864991 TI - [Complications during implantation of umbrella-type cava filter]. PMID- 9864992 TI - [Hemostasis of the gallbladder bed by using muscle flap with vascular pedicle]. PMID- 9864993 TI - [Hydatid echinococcus of the thoracic aorta]. PMID- 9864994 TI - [Complications of Meckel's diverticulum]. PMID- 9864995 TI - [Morphofunctional characteristics of monocytes. Significance of nucleolar system studies (Review of literature)]. PMID- 9864996 TI - [Hemopoiesis in chronic myelomonocytic leukemia]. AB - Hemopoiesis and functional activity of monocytes were assessed in 30 patients with chronic myelomonocytic leukemia (CMML). Changes in haemogram in CMML are characterized by anemia, thrombocytopenia, normal or increased count of leukocytes with absolute monocytosis and increased count of young granulocytes. Bone marrow hemopoiesis is characterized by enhanced proliferation of cellular elements of granulocytopoiesis and monocytes, suppressed erythropoiesis and megakaryocytopoiesis. Dysplastic changes are most expressed in the erythrocytic stem cells. Studies of lysosomal enzymes showed their increased activity in monocytes of CMML patients. PMID- 9864997 TI - [Cytological diagnosis of vascular neoplasms in the lung]. PMID- 9864998 TI - [Noninvasive method for glycemia detection]. PMID- 9864999 TI - [Diagnostic and prognostic significance of biochemical indicators in colorectal neoplasms]. AB - Activities of some enzymes and content of medium-weight molecules in patients with colorectal cancer were studied in order to assess the diagnostic value of these parameters for detection of tumors in the large intestine and development of endogenous intoxication after surgery and for prediction of remote results of treatment. Increased activities of creatine phosphokinase and hexokinase is typical of tumor growth, whereas increased activities of alkaline phosphatase and gamma-glutamyl transpeptidase are observed only in metastases to the liver. The role of medium-weight molecules in the diagnosis of endogenous intoxication during the early postoperative period is shown. The content of these molecules in the sera increases 3 days before clinical manifestation of endotoxicosis, when the traditional parameters are virtually normal. PMID- 9865000 TI - [Cooperation between clinic and laboratory: purpose, philosophy, and practice]. PMID- 9865001 TI - [Computer neural networks in clinical laboratory diagnosis]. PMID- 9865002 TI - [Laboratory of rapid diagnosis (validation, purpose, and procedures)]. AB - The authors discuss the problem of decreasing the number of errors in clinical laboratory diagnosis using artificial "neuron network" in which subjective and half-empirical euristics are replaced with objective regulations based on quantitative and logic analysis. The proposed method was used to derive the decisive rules ensuring prediction of outcomes of abdominal surgery. Reliability of prognostic table reflecting the new decisive rules in common form is confirmed by clinical data. PMID- 9865003 TI - [Material for external assessment of quality of glucose analysis]. AB - 9,10-bismethyltriphenylphosphoniumanthracene iodide (BPA) in concentrations 1.25 3.75 mmole/liter stabilizes blood serum and preserves glucose in it for 990 days at 4-8 degrees C. For preparing reference material, 200 ml human or animal serum is added to 800 ml 3.125 mM BPA solution, mixed, exposed for 24 h at 4-8 degrees C, centrifuged, the resultant preparation is stored at the same temperature and used as reference material for assessing the quality of glucose measurements by o toluidine method at biochemical laboratories. The reference material is fit for tests for at least 990 days. PMID- 9865004 TI - [Clinical chemistry in laboratory medicine in Europe. The past, present, and future]. PMID- 9865007 TI - [General pathology-theory of clinical medicine]. PMID- 9865008 TI - [Modern policy of anti-helicobacter therapy of ulcer disease]. PMID- 9865009 TI - [Tropical malaria]. PMID- 9865010 TI - [Role of melatonin in human body]. PMID- 9865011 TI - [Opportunities of coronary heart disease rehabilitation results objective assessment after coronary artery bypass surgery]. PMID- 9865012 TI - [Role of thrombolytic therapy in the treatment of patients with unstable angina]. AB - Thrombolytic therapy (TT) was performed in 39 of 81 patients with unstable angina pectoris (UAP) resistant to conventional treatment. TT was found to improve prognosis of UAP, to quicken UAP stabilization. Predictors of UAP unfavorable course were discovered. TT is indicated in patients suffering from resistant UAP with markers of poor prognosis. PMID- 9865013 TI - [Therapeutic plasmapheresis and alpha-tocopherol in patients with coronary heart disease]. AB - 46 patients with coronary heart disease with hypercholesterolemia were exposed to therapeutic plasmapheresis (TP) in combination with alpha-tocopherol treatment (AT). The results of 3-month follow-up with assessment of the clinical status, lipid spectrum, lipid peroxidation, concentration of ceruloplasmin indicated high hypolipidemic effectiveness of TP 2-3 weeks after the treatment as shown by inhibition of lipid peroxidation and antioxidant system. The addition of AT prolonged the hypolipidemic effect of TP, promoted optimization of plasma antioxidant potential (a rise in HDL, stabilization of ceruloplasmin levels). PMID- 9865014 TI - [Cardiac rhythm and conduction disorders in polymyositis and dermatomyositis]. AB - To assess condition of the heart in polymyositis (PM) and dermatomyositis (DM) in terms of defects in the rhythm and conductivity, we carried out ECG, 24-hour ECG monitoring, EIA measurements of KPK MB-fraction and myoglobin, echocardiography and Doppler echocardiography. Maximal frequency of arrhythmia presenting as supraventricular and ventricular extrasystole occurred in patients with acute PM/DM. Blockade of the bundle of His left anterior branch was detected most frequently in patients with acute and subacute PM-DM and in those with high serum levels of KPK MB-faction and myoglobin. Frequency of rhythmic and conductivity disorders seems unrelated to the disease duration. We registered no alterations at rest either in intracardiac hemodynamics or myocardial contractility. The study of these parameters under exercise test may provide objective information on early defects in myocardial contractility. PMID- 9865015 TI - [Adequate therapy of intestinal dysbacteriosis in children]. PMID- 9865016 TI - [Pharmacotherapeutical management of bronchial asthma]. AB - The analysis of the prescriptions for bronchial asthma (BA) patients in outpatient practice was made using data base created at the Regional Fund of Obligatory Health Insurance in the Sverdlovsk region with consideration of GINA principles of BA stepped care. The real structure of prescriptions was compared with the pattern drug official list for asthma care. The cost of each BA care step was calculated on the base of the computer programs. The analysis showed low quality of BA care in outpatient practice. The created computer programs are very useful for the prescriptions structure monitoring, cost-effect analysis and pharmacotherapeutical management of the diseases. PMID- 9865017 TI - [Hemodynamic chronosensitivity to metopress-retard in patients with stage ii hypertension]. AB - Time-dependent effects of metopress-retard (TEVA, Israel) were studied in 38 patients with stage II hypertension who were randomized to two groups, 19 patients each. Group 1 received 100 mg of metopress-retard at 8 a.m., group 2--at 8 p.m. Before the drug was taken and each 2 hours for 12 running hours after its 100 mg intake hemodynamic parameters were measured non-invasively. It was found that metopress-retard had no negative chronotropic and more strong hypotensive effects when taken in the evening (at 20.00). PMID- 9865018 TI - [Clinical trial of heptral in patients with chronic diffuse liver disease with intrahepatic cholestasis syndrome]. AB - Heptral (S-adenosine-L-methionine) was given to 32 patients with chronic diffuse diseases of the liver and intrahepatic cholestasis. 16 of them had primary biliary cirrhosis (PBC). Phase I of the treatment lasted 16 days when the drug was injected intravenously in a dose 800 mg/day. It was followed by phase 2--1600 mg/day taken for 16 days. A response was registered in the majority of patients. They had relieved symptoms of asthenia, skin pruritus, jaundice. The patients with liver cirrhosis and chronic hepatitis exhibited a statistically significant fall in ALT, AST and GGTP. PBS patients showed insignificant lowering of cholesterol, bilirubin. No resistance was noted in repeated courses. Heptral tolerance was satisfactory. PMID- 9865021 TI - [Hereditary membrane-deficiency hemolytic anemias]. PMID- 9865020 TI - [Difficulties in diagnosis of extrapulmonary tuberculosis running with hyperthermia and cytopenia]. PMID- 9865022 TI - [Automated profiloesophagotonometry in diagnosis of cardial insufficiency in patients with hiatal hernia]. PMID- 9865023 TI - [Jaundice]. PMID- 9865024 TI - [Role of I.M. Sechenov Moscow Medical Academy clinics in development of higher medical education]. PMID- 9865025 TI - [Ismar Boas (1858-1938)-founder of clinical gastroenterology]. PMID- 9865026 TI - [Lev Tolstoi's opinion about medicine and medical profession (part I)]. PMID- 9865027 TI - The concepts of language and the language of concepts. PMID- 9865028 TI - Concepts, knowledge and language in healthcare information systems: follow-up 30 months later. AB - The paper reviews the last IMIA-WG6 recommendations, issued along two themes: (1) how to better share common results and (2) how to conduct new research and development efforts. Some of the major discrepancies in the field are reviewed. The paper intends to offer insight into two areas, first, in the issue of knowledge representation and, second, in the science of meaning, incorporating the broad intersection of computational linguistics, semantics, conceptual approach, and denotation. The Ogden-Richards meaning triangle is used as a convenient organizing principle to maximize the scope of understanding among the different schools, particularly in areas of linguistics and semantics on the one side and denotation and conceptual approach on the other. In conclusion, there is a need for clarification and for more scientific pragmatism. PMID- 9865029 TI - Alternative ways for knowledge collection, indexing and robust language retrieval. AB - Definitions are provided of the key entities in knowledge representation for Natural Language Processing (NLP). Starting from the words, which are the natural components of any sentence, both the role of expressions and the decomposition of words into their parts are emphasized. This leads to the notion of concepts, which are either primitive or composite depending on the model where they are created. The problem of finding the most adequate degree of granularity for a concept is studied. From this reflection on basic Natural Language Processing components, four categories of linguistic knowledge are recognized, that are considered to be the building blocks of a Medical Linguistic Knowledge Base (MLKB). Following on the tracks of a recent experience in building a natural language-based patient encoding browser, a robust method for conceptual indexing and query of medical texts is presented with particular attention to the scheme of knowledge representation. PMID- 9865030 TI - The distinction between linguistic and conceptual semantics in medical terminology and its implication for NLP-based knowledge acquisition. AB - Natural language understanding systems have to exploit various kinds of knowledge in order to represent the meaning behind texts. Getting this knowledge in place is often such a huge enterprise that it is tempting to look for systems that can discover such knowledge automatically. We describe how the distinction between conceptual and linguistic semantics may assist in reaching this objective, provided that distinguishing between them is not done too rigorously. We present several examples to support this view and argue that in a multilingual environment, linguistic ontologies should be designed as interfaces between domain conceptualizations and linguistic knowledge bases. PMID- 9865031 TI - Evaluating natural language processors in the clinical domain. AB - Evaluating natural language processing (NLP) systems in the clinical domain is a difficult task which is important for advancement of the field. A number of NLP systems have been reported that extract information from free-text clinical reports, but not many of the systems have been evaluated. Those that were evaluated noted good performance measures but the results were often weakened by ineffective evaluation methods. In this paper we describe a set of criteria aimed at improving the quality of NLP evaluation studies. We present an overview of NLP evaluations in the clinical domain and also discuss the Message Understanding Conferences (MUC) [1-4]. Although these conferences constitute a series of NLP evaluation studies performed outside of the clinical domain, some of the results are relevant within medicine. In addition, we discuss a number of factors which contribute to the complexity that is inherent in the task of evaluating natural language systems. PMID- 9865032 TI - Conceptual graph grammar--a simple formalism for sublanguage. AB - There are a wide variety of computer applications that deal with various aspects of medical language: concept representation, controlled vocabulary, natural language processing, and information retrieval. While technical and theoretical methods appear to differ, all approaches investigate different aspects of the same phenomenon: medical sublanguage. This paper surveys the properties of medical sublanguage from a formal perspective, based on detailed analyses cited in the literature. A review of several computer systems based on sublanguage approaches shows some of the difficulties in addressing the interaction between the syntactic and semantic aspects of sublanguage. A formalism called Conceptual Graph Grammar is presented that attempts to combine both syntax and semantics into a single notation by extending standard Conceptual Graph notation. Examples from the domain of pathology diagnoses are provided to illustrate the use of this formalism in medical language analysis. The strengths and weaknesses of the approach are then considered. Conceptual Graph Grammar is an attempt to synthesize the common properties of different approaches to sublanguage into a single formalism, and to begin to define a common foundation for language-related research in medical informatics. PMID- 9865033 TI - The nature of lexical knowledge. AB - This paper considers the nature of lexical knowledge and its role in language and information processing. The lexicon is the central component of language and plays a pivotal role in current linguistic theory [3, 4] and, increasingly, in natural language processing systems [5-7]. The lexicon embodies information about the lexical items of the language and serves as the foundation for morphologic, syntactic, and semantic processing. The differences as well as commonalities among dictionaries, thesauri, and lexicons are discussed, and distinctions between words, lexical items, and terms are drawn. Next, the scope and content of the SPECIALIST lexicon are presented, followed by a discussion of certain writing conventions that can be troublesome for text processing applications. One approach to handling orthographic and other lexical variation is discussed in a section that reports on the design and implementation of the SPECIALIST lexical programs. The paper concludes with a discussion of controlled terminologies for the medical domain. Throughout the discussion, examples are drawn from the SPECIALIST lexicon and from the other UMLS knowledge sources [8, 9]. PMID- 9865034 TI - Modeling concepts in medicine for medical language understanding. AB - Over the past two decades, the construction of models for medical concept representation and for understanding of the deep meaning of medical narrative texts have been challenging areas of medical informatics research. This review highlights how these two inter-related domains have evolved, emphasizing aspects of medical modeling as a tool for medical language understanding. A representation schema, which balances partially but accurately with complete but complex representations of domain-specific knowledge, must be developed to facilitate language understanding. Representative examples are drawn from two major independent efforts undertaken by the authors: the elaboration and the subsequent adjustment of the RECIT multilingual analyzer to include a robust medical concept model, and the recasting of a frame-based interlingua system, originally developed to map equivalent concepts between controlled clinical vocabularies, to invoke a similar concept model. PMID- 9865035 TI - Metaphrase: an aid to the clinical conceptualization and formalization of patient problems in healthcare enterprises. AB - Patient descriptors, or "problems," such as "brain metastases of melanoma" are an effective way for caregivers to describe patients. But most problems, e.g., "cubital tunnel syndrome" or "ulnar nerve compression," found in problem lists in an Electronic Medical Record (EMR) are not comparable computationally--in general, a computer cannot determine whether they describe the same or a related problem, or whether the user would have preferred "ulnar nerve compression syndrome." Metaphrase is a scalable, middleware component designed to be accessed from problem-manager applications in EMR systems. In response to caregivers' informal descriptors it suggests potentially equivalent, authoritative, and more formally comparable descriptors. Metaphrase contains a clinical subset of the 1997 UMLS Metathesaurus and some 10,000 "problems" from the Mayo Clinic and Harvard Beth Israel Hospital. Word and term completion, spelling correction, and semantic navigation, all combine to ease the burden of problem conceptualization, entry and formalization. PMID- 9865036 TI - From text to knowledge: a unifying document-centered view of analyzed medical language. AB - Although medical language processing (MLP) has achieved some success, the actual use and dissemination of data extracted from free text by MLP systems is still very limited. We claim that the adoption of an 'enriched-document' paradigm (or 'document-centered' view) can help to address this issue. We present this paradigm and explain how it can be implemented, then discuss its expected benefits both for end-users and MLP researchers. PMID- 9865038 TI - The SNOMED DICOM microglossary: controlled terminology resource for data interchange in biomedical imaging. AB - This paper describes an authoritative, non-proprietary information resource that provides an efficient mechanism for embedding specialized clinical knowledge into the design of healthcare telecommunications systems. The resource marries two types of data interchange standards, a message/electronic-document standard and a terminology standard. In technical terms, it is part protocol and part database. Industry, academia, professional specialty societies, and the federal government participated in its development. The development of multi-specialty content has broadly engaged biomedical domain experts to an unprecedented degree in voluntary, non-proprietary message/document-standards development. The resource is the SNOMED DICOM Microglossary (SDM), a message-terminology (or document content) mapping resource. The message/electronic-document standard is DICOM (Digital Imaging and Communications in Medicine). The terminology standard is SNOMED, (Systematized Nomenclature of Human and Veterinary Medicine). The SDM specifies the mapping of multi-specialty imaging terminology from SNOMED to DICOM data elements. DICOM provides semantic constraints and a framework for discourse that are lacking in SNOMED. Thus the message standard and the computer-based terminology both depend upon and complete each other. The combination is synergistic. By substitution of different templates of specialty terminology from the SDM, a generic message template, such as the DICOM Visible Light (Color Diagnostic) Image or the DICOM Structured Reporting specification can be reconfigured for diverse applications. Professional societies, with technical assistance from the College of American Pathologists, contribute and maintain their portions of the terminology, and can use SDM templates and term lists in clinical practice guidelines for the structure and content of computer-based patient records. PMID- 9865037 TI - Desiderata for controlled medical vocabularies in the twenty-first century. AB - Builders of medical informatics applications need controlled medical vocabularies to support their applications and it is to their advantage to use available standards. In order to do so, however, these standards need to address the requirements of their intended users. Over the past decade, medical informatics researchers have begun to articulate some of these requirements. This paper brings together some of the common themes which have been described, including: vocabulary content, concept orientation, concept permanence, nonsemantic concept identifiers, polyhierarchy, formal definitions, rejection of "not elsewhere classified" terms, multiple granularities, multiple consistent views, context representation, graceful evolution, and recognized redundancy. Standards developers are beginning to recognize and address these desiderata and adapt their offerings to meet them. PMID- 9865039 TI - Semantic definition of disorders in version 3 of the Read Codes. AB - The disorder chapter of Version 3 of the Read codes is a rich source of clinically derived terminology. The file structure has been designed to meet a clinical need to support both enumerated and compositional taxonomy. This requirement coupled with the maintenance of multiple classification necessitates a mechanism to identify consistent hierarchical placement and equivalence between concepts. The early work on the semantic definition of the disorder chapter, to support these requirements, is outlined and the attributes that have been found to be important are presented. We also describe different categories of completeness of definition that have been identified, and the scope of those that are likely to remain incompletely characterised. PMID- 9865040 TI - Exploring the ontology of surgical procedures in the Read Thesaurus. AB - The Read Thesaurus is a comprehensive user-led clinical vocabulary developed from earlier, and structurally simpler, versions of the Read Codes, with substantial input from United Kingdom health care professionals. A constituent template table underpins a range of functions, including semantic definition of concepts using object-attribute-value triples. Concept representation for surgical procedures has been investigated by a number of groups and a standard European structure has been proposed. Over 50% of the surgical procedures in the Read Thesaurus have been fully characterised using a number of attributes each with a defined concept field. We report progress to date and, based on our large-scale experience, examine the applicability of the European model to a user-defined terminology. PMID- 9865041 TI - Scalable methodologies for distributed development of logic-based convergent medical terminology. AB - As the size and complexity of medical terminologies increase, terminology modelers are increasingly hampered by lack of tools and methods to manage the development process. This paper presents our use and ongoing evaluation of a description-logic classifier to support cognitive scalability of the underlying terminology and our enhancements to that classifier to support concurrent development utilizing semantics-based concurrency control methods. Our enhancements, collectively referred to as the Galapagos, consist of several applications that take locally-developed terminology enhancements from multiple sites, identify conflicting design decisions, support the modelers' reconciliation of the conflicting designs, and efficiently disseminate updates tailored for locally enhanced terminologies. We have tested our ideas through concurrent evolutionary enhancement of SNOMED International at three Kaiser Permanente regions and the Mayo Clinic. We have found that the underlying environment has met our design objectives, and supports semantic-based concurrency control, and identification and resolution of conflicting design decisions. PMID- 9865042 TI - Linking a medical vocabulary to a clinical data model using Abstract Syntax Notation 1. AB - We have created a clinical data model using Abstract Syntax Notation 1 (ASN. 1). The clinical model is constructed from a small number of simple data types that are built into data structures of progressively greater complexity. Important intermediate types include Attributes, Observations, and Events. The highest level elements in the model are messages that are used for inter-process communication within a clinical information system. Vocabulary is incorporated into the model using BaseCoded, a primitive data type that allows vocabulary concepts and semantic relationships to be referenced using standard ASN. 1 notation. ASN. 1 subtyping language was useful in preventing unbounded proliferation of object classes in the model, and in general, ASN.1 was found to be a flexible and robust notation for representing a model of clinical information. PMID- 9865043 TI - Three types of IS-A statement in diagnostic classifications: three types of knowledge needed for development and maintenance. AB - Update mechanisms for diagnostic classifications should capture changes in medical knowledge but also allow for comparability across versions. This paper provides a basis for such a mechanism by describing types of IS-A statement and types of knowledge used in the construction of diagnostic classifications. Three types of IS-A statement are used: 'A is by definition a B', 'A is probably a B' and 'A is in theory necessarily a B'. Each relates to a different type of knowledge: knowledge of linguistic conventions, of probabilities, and of empirical theories and their status, respectively. Consequently, the development and maintenance of diagnostic classifications requires a collaboration of medical terminologists and medical scientists. The role of the latter is especially important during updating. Updating is necessitated by changing probabilities and by the introduction or changing status of empirical theories. The linguistic notion of hyponymy oversimplifies the issue. PMID- 9865044 TI - Understanding, navigating and communicating knowledge: issues and challenges. AB - This paper presents a psychological perspective on key issues related to medical vocabularies. There have been rapid advances in the development of computer technology underlying medical information systems. However, in keeping with technological progress, we must also take into account advances in our understanding of human behaviour and learn from failures in human performance. A central issue examined in this paper is the extent to which we can develop generic vocabularies that are also flexible and adaptable to specific situations. Empirical research indicates that variability in human performance is much greater than what current medical classifications take into account. A related challenge is that of how to best develop vocabularies that meet the needs of users. Based on theoretical perspectives and research emerging from the domain of cognitive psychology, we suggest that an understanding of the cognitive mechanisms underlying the comprehension and application of terminology is required. It is argued that rather than beginning with highly specified terminologies, i.e. the normative approach, we might instead begin by examining the natural context of how health care workers acquire, understand and negotiate knowledge in practice. PMID- 9865045 TI - How groups co-ordinate their concepts and terminology: implications for medical informatics. AB - Conceptual and terminological systems are established and maintained by the communities who use them. This paper reports experiments which investigate the role of communication and interaction in the process. The experiments show that isolated pairs of communicators and virtual communities of interacting pairs naturally converge on their own conceptual and terminological systems when confronted with a common task. The results also indicate that the system converged on is optimal for that particular group engaged in that particular task. These findings are discussed in relation to the increasing use of tightly coordinated medical teams and its implications for getting them to adopt standardized medical terminologies. PMID- 9865046 TI - Evaluation of a semantic data model for chest radiology: application of a new methodology. AB - An essential step toward the effective processing of the medical language is the development of representational models that formalize the language semantics. These models, also known as semantic data models, help to unlock the meaning of descriptive expressions, making them accessible to computer systems. The present study tries to determine the quality of a semantic data model created to encode chest radiology findings. The evaluation methodology relied on the ability of physicians to extract information from textual and encoded representations of chest X-ray reports, whilst answering questions associated with each report. The evaluation demonstrated that the encoded reports seemed to have the same information content of the original textual reports. The methodology generated useful data regarding the quality of the data model, demonstrating that certain segments were creating ambiguous representations and that some details were not being represented. PMID- 9865047 TI - Cooperative knowledge evolution: a construction-integration approach to knowledge discovery in medicine. AB - In this paper, we perform a cognitive analysis of knowledge discovery processes. As a result of this analysis, the construction-integration theory is proposed as a general framework for developing cooperative knowledge evolution systems. We thus suggest that for the acquisition of new domain knowledge in medicine, one should first construct pluralistic views on a given topic which may contain inconsistencies as well as redundancies. Only thereafter does this knowledge become consolidated into a situation-specific circumscription and the early inconsistencies become eliminated. As a proof for the viability of such knowledge acquisition processes in medicine, we present the IDEAS system, which can be used for the intelligent documentation of adverse events in clinical studies. This system provides a better documentation of the side-effects of medical drugs. Thereby, knowledge evolution occurs by achieving consistent explanations in increasingly larger contexts (i.e., more cases and more pharmaceutical substrates). Finally, it is shown how prototypes, model-based approaches and cooperative knowledge evolution systems can be distinguished as different classes of knowledge-based systems. PMID- 9865048 TI - Thesauri and formal classifications: terminologies for people and machines. AB - Terminologies are now software. They are key components of the integration of electronic patient records, decision support systems and information retrieval systems. To be used as software, the different types of content in traditional terminologies must be separated, which we term here: conceptual, linguistic, inferential and pragmatic. The conceptual knowledge at the heart of the terminology needs to be expressed formally in order to provide a dependable framework for the other types of knowledge. Information left implicit in most existing coding and classification systems must be made explicit. The test of the resulting terminologies is how well they support software for key functions: including data entry, information retrieval, mediation, indexing, and authoring. PMID- 9865049 TI - Formal classification of medical concept descriptions: graph-oriented operators. AB - A crucial component of a medical concept representation system is the classifier. It requires features that are not sufficiently supported by current logic based formalisms like description logics and conceptual graphs. Those features are, for instance, the representation of partitive and spatial relations and their impact on subsumption. This paper introduces graph oriented classification operators for a concept representation language with normal forms. Emphasis is on the separation of generic and partitive relations and on the mutual interdependence of subsumption and part-whole. For that purpose operators are given for formal subsumption, formal part-whole, subsumptive part-whole and part-sensitive subsumption. These operators are based on the formal structure of concept descriptions and on explicitly introduced generic and partitive relationships between their constituents. PMID- 9865050 TI - Rationale and design considerations for a semantic mediator in health information systems. AB - Rapid development of community health information networks raises the issue of semantic interoperability between distributed and heterogeneous systems. Indeed, operational health information systems originate from heterogeneous teams of independent developers and have to cooperate in order to exchange data and services. A good cooperation is based on a good understanding of the messages exchanged between the systems. The main issue of semantic interoperability is to ensure that the exchange is not only possible but also meaningful. The main objective of this paper is to analyze semantic interoperability from a software engineering point of view. It describes the principles for the design of a semantic mediator (SM) in the framework of a distributed object manager (DOM). The mediator is itself a component that should allow the exchange of messages independently of languages and platforms. The functional architecture of such a SM is detailed. These principles have been partly applied in the context of the HELIOS object-oriented software engineering environment. The resulting service components are presented with their current state of achievement. PMID- 9865051 TI - Concept-oriented standardization and statistics-oriented classification: continuing the classification versus nomenclature controversy. AB - Nowadays, most activities on controlled medical vocabularies focus on the provision of a sufficient atomic-level granularity for representing clinical data. Amongst others, clinical vocabularies should be concept oriented, compositional and should also reject "Not Elsewhere Classified". We strongly share the opinion that there is a need to deal with serious deficits of existing manually created vocabularies and with new demands for computer-based advanced processing and exchange of medical language data. However, we do not share the opinion that methodological requirements like observational and structural comparability needed for sound statistics should not be included in desiderata of controlled medical vocabularies. Statistical-oriented classifications are not developed for representing detailed clinical data but for providing purpose dependent classes where cases of interest are assigned uniquely. Either statistical classifications are not included into the set of controlled medical vocabularies in the sense of Cimino, or his desiderata are misleading. We argue that statistical classifications should be linked to (formal) concept systems, but again this linkage does not change their different natures. With this article we continue the "classification versus nomenclature" controversy referring to Cote. PMID- 9865052 TI - Domain ontologies in software engineering: use of Protege with the EON architecture. AB - Domain ontologies are formal descriptions of the classes of concepts and the relationships among those concepts that describe an application area. The Protege software-engineering methodology provides a clear division between domain ontologies and domain-independent problem-solvers that, when mapped to domain ontologies, can solve application tasks. The Protege approach allows domain ontologies to inform the total software-engineering process, and for ontologies to be shared among a variety of problem-solving components. We illustrate the approach by describing the development of EON, a set of middleware components that automate various aspects of protocol-directed therapy. Our work illustrates the organizing effect that domain ontologies can have on the software-development process. Ontologies, like all formal representations, have limitations in their ability to capture the semantics of application areas. Nevertheless, the capability of ontologies to encode clinical distinctions not usually captured by controlled medical terminologies provides significant advantages for developers and maintainers of clinical software applications. PMID- 9865053 TI - Standards to support development of terminological systems for healthcare telematics. AB - The Technical Committee on "Medical Informatics" of the European Committee for Standardization (CEN/TC251) is supporting developers of terminological systems in healthcare by a series of standards. The dream of "universal" coding system was abandoned in favor of a coherent family of terminologies, diversified according to tasks; two ideas were introduced: (1) the "categorical structure", i.e. a model of semantic categories and their relations within a subject field and (2) the "cross-thesaurus", i.e. a system of descriptors to build a systematic representation (called here "dissection") for each terminological phrase, coherent across diverse terminologies on a given subject field. The goal is to assure coexistence and interoperability (and reciprocal support for development and maintenance) to three generations of systems: (1) traditional paper-based systems (first generation); (2) compositional systems built according to a categorical structure and a cross-thesaurus (second generation) and (3) formal models (third generation). Various scenarios are presented, on the exploitation of computer-based terminological systems. The idea of "operational meaning" of terminological phrases within administrative and organizational contexts and the idea of "task-oriented details" are also introduced, to justify and exploit design constraints on terminological systems. PMID- 9865054 TI - Towards cooperative patient management through organizational and medical ontologies. AB - Within knowledge and data engineering a new research paradigm is emerging based on the Multi-Agent System (MAS) architectural framework, allowing human and software agents to interoperate and thus cooperate within common application areas. In such a framework, knowledgeable agents of heterogeneous nature, that possess diverse but at least partially compatible or inter-translatable conceptual views, or ontologies, modeling both their own expertise and the external environment, make somehow available their information resources or problem-solving abilities for cooperative processes addressing the construction of a new agent or the achievement of some common goal through a correlated execution of tasks. In this paper, we restrict our analysis to the case of an organization of cognitive agents, illustrated with examples from a prototypical healthcare MAS, that is, a so-called Distributed Healthcare Information System (D HIS). The prototype makes use of an ontological library written in the standard language Ontolingua. An ongoing application of the methodology to the main problem of Clinical Practice Guidelines (GLs) computer-based dissemination and enforcement is described. PMID- 9865055 TI - [Recent progress in the treatment of ischemic heart disease in the elderly]. PMID- 9865056 TI - [Expression and structure of senescence marker protein-30 (SMP30) and its physiological function]. PMID- 9865057 TI - [Esophageal diseases in the elderly patients]. PMID- 9865059 TI - [Diabetes mellitus in the elderly]. PMID- 9865058 TI - [Thyroid disease in ageing]. PMID- 9865060 TI - [Clinical practice of dementia of the elderly]. PMID- 9865061 TI - [Cerebrovascular diseases in patients with arteriosclerotic aortic aneurysm and arteriosclerosis obliterans studied by carotid ultrasonography and by computed tomography of the brain]. AB - To estimate the incidence of cerebrovascular diseases in arteriosclerotic aortic aneurysm (AA) and arteriosclerosis obliterans (ASO) and their characteristics, 92 patients with AA and 102 patients with ASO were studied with carotid ultrasonography and brain computed tomography (CT), and were compared with 49 patients with hypertension. The mean ages of the patients were 70 to 72 years old and all were male. Hypertension was common in the AA group: diabetes and cigarette smoking were common in the ASO group. Carotid plaque lesions seen on ultrasonography were significantly more common in the AA (66%) and ASO (85%) groups than in the patients with hypertension (39%). The mean carotid diameter was significantly greater in the AA group than in the other two groups. The mean wall thicknesses in the AA and ASO groups were greater than in the patients with hypertension. Computed tomography showed that low-density areas were also common in the AA group (56%) and ASO (53%) groups than in the patients with hypertension (24%). Most of the low-density areas were thought to be lacunae or dilated perivascular spaces in the subinsula and putamen. Moderately and highly severe periventricular lucencies were also common in the AA group. These findings indicate that carotid changes, seen in AA and ASO, reflected the characteristics of arterial lesions, and that arteriolosclerotic lesions were common in patients with AA and ASO. Therefore, cerebrovascular diseases should be taken into account in the management of patients with AA and ASO. PMID- 9865062 TI - [Serum globulin levels and activities of daily living in centenarians]. AB - Serum globulin levels in 101 centenarians (77 women, 24 men) were analyzed by zonal electrophoresis and immunoglobulin assays. Six subjects showed an M band in electrophoretograms. Five M bands were immunoglobulin G (IgG) type and one was IgA. One subject was suspected to suffer from IgG-type multiple myeloma, and the others were thought to have essential (benign) monoclonal gammopathy. The relationship between serum globulin levels and activities of daily living (ADL) was studied in remaining 95 subjects. There were 21 rank-J (free-living), 25 rank A (unable to go outside without help), 23 rank-B (bedridden but able to sit on the bed), and 26 rank-C (completely bedridden) centenarians. The mean serum alpha 1-, alpha 2-, and beta-globulin level in the rank-J centenarians was at the upper limit of the normal adult range. The mean alpha 1-globulin levels in rank-B (3.3 +/- 0.4%) and C (3.4 +/- 0.5%) subjects were significantly higher than those in rank-J (2.9 +/- 0.3%) centenarians. The mean serum gamma-globulin level of rank-J centenarians (16.9 +/- 3.7%) was within the normal range. The mean gamma-globulin levels in rank-B (21.0 +/- 2.7%) and C (22.8 +/- 4.9%) subjects were significantly higher than that in rank-J centenarians. The mean immunoglobulin G concentrations in rank-B (1.620 +/- 304 mg/dl) and C (1.720 +/- 392 mg/dl) subjects were significantly higher than those in rank-J centenarians (1.320 +/- 361 mg/dl) and in normal adults (1.150 +/- 235 mg/dl). IgA concentrations in all centenarians were higher than in normal controls. There was no significant difference in IgA or IgM concentration between rank-J subjects and those in other ranks. Serum globulin levels measured with electrophoresis, and the immunoglobulin concentrations, can be valuable indicators of the ability to perform activities of daily living, and may be useful predictors of subclinical diseases or morbidity in centenarians. PMID- 9865063 TI - [Assessment of localized hypertrophy in the basal part of the interventricular septum in the elderly]. AB - The basal part of the interventricular septum (IVS) is known to show different hypertrophic features from those observed in the other parts of the left ventricular wall. These are considered to reflect physiological changes that occur with normal aging. However, these changes have not been carefully evaluated, and their clinical significance has not been defined. We assessed these changes echocardiographically. The subjects were patients at least 70 years of age in whom localized hypertrophy in the basal part of the IVS was seen during the whole cardiac cycle on echocardiography. The prevalence was 6.3% among 96 consecutively studied patients. All 6 patients had a history of hypertension. Echocardiographic findings were as follows: 1) the left atrium was mildly or moderately dilated, 2) there was no evidence of either dilatation or narrowing of the left ventricular cavity, 3) the left ventricular wall motion appeared normal and indices of systolic function were within normal limits in all subjects except one who had a history of myocardial infarction, 4) the angle formed by the aorta and the IVS averaged 106.7 degrees (range: 95 to 120 degrees), 5) Doppler examination showed increases in the ratio of the peak flow velocity during atrial systole to the peak flow velocity early in diastole, and 6) prolongation of the deceleration time of the flow velocity early in diastole. The last of these findings suggested left ventricular diastolic dysfunction, but peak flow velocity at the left ventricular outflow tract was normal. There was no evidence of stenosis of the left ventricular outflow tract. Localized hypertrophy in the basal part of the IVS in elderly patients could be a type of cardiac hypertrophy caused by hypertension. On echocardiography, the basal part of the IVS seemed to protrude toward the left ventricular cavity, but there was no evidence of stenosis in the left ventricular outflow tract. PMID- 9865064 TI - [Bilateral homonymous hemianopsia with atrial fibrillation]. AB - We encountered an 80-year-old man with sudden bilateral visual disturbance. When he was admitted to the hospital, his blood pressure was 138/70 mmHg, and an ECG revealed atrial fibrillation. Neurological examination showed only bilateral homonymous hemianopsia with no nystagmus or impairment of eye movement. He did not have paralysis cerebellar ataxia, or speech disturbance. Therefore, there was no evidence of obstruction of the thalamogeniculate or thalamoperforating artery. Magnetic resonance imaging of the brain showed cerebral infarctions in both occipital lobes. Perimetry showed bilateral homonymous hemianopsia; the left side of the macula was spared. The lack of neurological deficit other than bilateral homonymous hemianopsia indicates that only the right and left cortical branches of the posterior cerebral artery were occluded. PMID- 9865065 TI - [Study on stimulatory mechanism of inflammatory bone resorption induced by Porphyromonas gingivalis]. PMID- 9865066 TI - [Guanine nucleotide-exchange proteins (GEP) for ADP-ribosylation factors (ARF)]. PMID- 9865067 TI - [Common antigenicity of lipopolysaccharides between Rickettsiae and Proteus in the Weil-Felix test]. PMID- 9865068 TI - [Molecular epidemiology and pathogenicity of Clostridium difficile]. PMID- 9865069 TI - [Recent progress in DNA diagnosis of bacteria]. PMID- 9865070 TI - [History of nomenclature of Salmonella species and the legitimate names]. PMID- 9865071 TI - Temporal inhibition in character identification. AB - Models of information processing tasks such as character identification often do not consider the nature of the initial sensory representation from which task relevant information is extracted. An important component of this representation is temporal inhibition, in which the response to a stimulus may inhibit, or in some cases facilitate, processing of subsequent stimuli. Three experiments demonstrate the existence of temporal inhibitory processes in information processing tasks such as character identification and digit recall. An existing information processing model is extended to account for these effects, based in part on models from the detection literature. These experiments also discriminate between candidate neural mechanisms of the temporal inhibition. Implications for the transient deficit theory of dyslexia are discussed. PMID- 9865072 TI - Directed attention prolongs the perceived duration of a brief stimulus. AB - Stelmach, Herdman, and McNeil (1994) suggested recently that the perceived duration for attended stimuli is shorter than that for unattended ones. In contrast, the attenuation hypothesis (Thomas & Weaver, 1975) suggests the reverse relation between directed attention and perceived duration. We conducted six experiments to test the validity of the two contradictory hypotheses. In all the experiments, attention was directed to one of two possible stimulus sources. Experiments 1 and 2 employed stimulus durations from 70 to 270 msec. A stimulus appeared in either the visual or the auditory modality. Stimuli in the attended modality were rated as longer than stimuli in the unattended modality. Experiment 3 replicated this finding using a different psychophysical procedure. Experiments 4-6 showed that the finding applies not only to stimuli from different sensory modalities but also to stimuli appearing at different locations within the visual field. The results of all six experiments support the assumption that directed attention prolongs the perceived duration of a stimulus. PMID- 9865073 TI - Estimation of height for persons in pictures. AB - We report three experiments in which subjects estimated the height of persons seen in pictures. In Experiment 1, 50 realistic photographs were used as targets. The correlation between mean estimated height and actual height was .92, and target's sex and the ratios of head to trunk, which included horizontal ratio (i.e., a ratio of head width to shoulder width) and vertical ratio (i.e., a ratio of head height to stature), were found to be effective on height estimates, although target's sex was more effective than ratios of head to trunk. In Experiments 2 and 3, the recognizability of target persons was reduced, respectively, by inverting the person photographs and by degrading the various features of the photographs. The main results were that (1) the correlation between mean estimated height and actual height was .88 for the inverted photographs and was .80, on average, for the degraded drawings; (2) target's sex was consistently influential in both experiments; (3) vertical ratio was effective for the inverted photographs and the mildly degraded drawings but not for the extremely degraded drawings in which only outlines of target persons were depicted; and (4) the estimation for the outlined pictures was likely to be influenced by fit of clothes to targets' bodies. From these findings, person stimuli were considered to provide a stable metric stick of visual space under naturalistic circumstances. PMID- 9865074 TI - The precision of velocity discrimination across spatial frequency. AB - The precision of velocity coding for moving stimuli of different spatial frequencies was assessed by measuring velocity discrimination thresholds for a 1 c/deg grating paired with a grating whose spatial frequency ranged from 0.25 to 4 c/deg and for grating pairs of the same spatial frequency (0.25, 1, and 4 c/deg). The gratings always moved upward, with velocities ranging from 0.5 to 16 deg/sec. Velocity discrimination was as precise for stimuli that varied in spatial frequency by +/- 2 octaves (0.25 vs. 1 c/deg and 4 vs. 1 c/deg) as for stimuli of the same spatial frequency, for specific ranges of velocity that depended on the spatial and, therefore, the temporal frequencies of the stimuli. Compared with a 1-c/deg grating, the perceived velocity of 4-c/deg gratings was about 1.3 times faster and that of 0.25-c/deg gratings was about 1.3 times slower. Although these perceived velocity biases imply variation of velocity-signal processing among spatial frequency channels, the discrimination results indicate that the motion sensing system can compare signals across different spatial frequency channels to make fine velocity discrimination within appropriate temporal frequency limits. PMID- 9865075 TI - A maximum motion technique for assessment of color vision defects. AB - Several luminance-matching methods, such as flicker fusion and the minimum motion technique (MMT), are capable of detecting certain forms of abnormal color vision. We present evidence that the heterochromatic fusion nystagmus (HFN) luminance matching technique can discriminate among normal trichromats, protanopes, and deuteranomals. The HFN luminance matching technique has the advantage that it provides a positive indication of isoluminance (maximization of motion) as opposed to the MMT and flicker fusion methods, which indicate isoluminance by the minimization of motion and flicker, respectively. We tested 16 normal trichromats, 6 protanopes, and 4 deuteranomals with the HFN technique. Results indicate that HFN is a useful tool for examining color vision. Because the HFN stimulus elicits reflexive eye movements (optokinetic nystagmus) that follow the apparent motion of the stimulus, HFN luminance matching will be particularly useful in animal and infant research, where other color vision tests are difficult to implement. PMID- 9865076 TI - The asynchronous discrete coding model: further tests with single-attribute stimuli. AB - Two experiments using a probe-RT paradigm showed that partial information about the size of a stimulus can influence response processes before complete size information is available. Contrary to the asynchronous discrete coding model, these results suggest that the perceptual system may transmit to the response system information that only partially specifies a single stimulus attribute. In combination with previous findings, these results also suggest that there are at least two dissociable forms of selective preparation for a given response. PMID- 9865077 TI - Tempo discrimination of musical patterns: effects due to pitch and rhythmic structure. AB - The purpose of this research was to investigate a set of factors that may influence the perceived rate of an auditory event. In a paired-comparison task, subjects were presented with a set of music-like patterns that differed in their relative number of contour changes and in the magnitude of pitch skips (Experiment 1) as well as in the compatibility of rhythmic accent structure with the arrangement of pitch relations (Experiment 2) Results indicated that, relative to their standard referents, comparison melodies were judged to unfold more slowly when they displayed more changes in pitch direction, greater pitch distances, and an incompatible rhythmic accent structure. These findings are suggested to stem from an imputed velocity hypothesis, in which people overgeneralize certain invariant relations that typically occur between melodic and temporal accent structure within Western music. PMID- 9865078 TI - The effects of attenuation of frequency segments on binaural localization of sound. AB - Perceived location of tonal stimuli d and narrow noise bands presented in two dimensional space varies in an orderly manner with changes in stimulus frequency. Hence, frequency has a referent in space that is most apparent during monaural listening. The assumption underlying the present study is that maximum sound pressure level measured at the ear canal entrance for the various frequencies serves as a prominent spectral cue for their spatial referents. Even in binaural localization, location judgments in the vertical plane are strongly influenced by spatial referents. We measured sound pressure levels at the left ear canal entrance for 1.0-kHz-wide noise bands, centered from 4.0 kHz through 10.0 kHz, presented at locations from 60 degrees through -45 degrees in the vertical plane; the horizontal plane coordinate was fixed at -90 degrees. On the basis of these measurements, we fabricated three different bandstop stimuli in which differently centered 2.0-kHz-wide frequency segments were filtered from a broadband noise. Unfiltered broadband noise served as the remaining stimulus. Localization accuracy differed significantly among stimulus conditions (p < .01). Where in the vertical plane most errors were made depended on which frequency segment was filtered from the broadband noise. PMID- 9865079 TI - Multiple-feature discrimination faster than single-feature discrimination within the same object? AB - In the present study, we investigated whether judging the presence of multiple features within an object would be superior to judging the presence of only one feature. Feature discriminability and the number of features to discriminate within an object were varied. Specific features were judged as present or absent. Results showed that judging the presence of two or three features was faster than judging the presence of the less discriminable of these two or three features alone (multiple-feature benefits). These findings suggest that relevant features within an object activate (prime) a decision or response in a parallel, asynchronous fashion based on discriminability (Miller, 1982a). The ability of a response priming model, a response mapping model, and a template model to account for multiple-feature benefits is discussed. PMID- 9865080 TI - New evidence for the zoom lens model using the RSVP technique. AB - A main prediction from the zoom lens model for visual attention is that performance is an inverse function of the size of the attended area. The "attention shift paradigm" developed by Sperling and Reeves (1980) was adapted here to study predictions from the zoom lens model. In two experiments two lists of items were simultaneously presented using the rapid serial visual presentation technique. Subjects were to report the first item he/she was able to identify in the series that did not include the target (the letter T) after he/she saw the target. In one condition, subjects knew in which list the target would appear, in another condition, they did not have this knowledge, having to attend to both positions in order to detect the target. The zoom lens model predicts an interaction between this variable and the distance separating the two positions where the lists are presented. In both experiments, this interaction was observed. The results are also discussed as a solution to the apparently contradictory results with regard to the analog movement model. PMID- 9865081 TI - Dual adaptation and adaptive generalization of the human vestibulo-ocular reflex. AB - In two experiments, we examined the possibility that the human vestibulo-ocular reflex (VOR) is subject to dual adaptation (the ability to adapt to a sensory rearrangement more rapidly and/or more completely after repeated experience with it) and adaptive generalization (the ability to adapt more readily to a novel sensory rearrangement as a result of prior dual adaptation training). In Experiment 1, the subjects actively turned the head during alternating exposure to a visual-vestibular rearrangement (target/head gain = 0.5) and the normal situation (target/head gain = 0.0). These conditions produced both adaptation and dual adaptation of the VOR but no evidence of adaptive generalization when tested with a target/head gain of 1.0. Experiment 2, in which exposure to the 0.5 gain entailed externally controlled (i.e., passive) whole body rotation, resulted in VOR adaptation but no dual adaptation. As in Experiment 1, no evidence of adaptive generalization was found. PMID- 9865082 TI - Dissociation of covert and overt spatial attention during prehension movements: selective interference effects. AB - In four experiments, the influence of distractor objects on the temporal evolution of the reach-to-grasp movement toward a target object (an apple) was examined. In the first experiment, the distractor was another apple, which moved laterally behind the target and occasionally changed direction toward the target, thus becoming the to-be-grasped object. In the second and third experiments, the distractor was a stationary piece of fruit, which sometimes became the to-be grasped object because of a change in illumination. The fourth experiment was a combination of the first two experiments. In all cases, selective interference effects on the transport and manipulation components were observed only when attention to the distractor was covert rather than overt. It is proposed that covert visuospatial attention selects information about distracting but potentially important stimuli, such that a registration of significance is accomplished without the need to process all available information. PMID- 9865083 TI - The effect of timing and spatial separation on the velocity of auditory apparent motion. AB - Previously, it was shown that the minimum conditions for the illusion of auditory apparent motion (AAM) depend on stimulus timing but not spatial separation. In the present experiment, the effects of stimulus timing and source separation on the perceived velocity of AAM were examined. Eight listeners estimated the velocity, duration, and distance traveled of AAM, using a no-modulus, magnitude estimation procedure. Four burst durations (25, 50, 100, and 300 msec), 10 stimulus onset asynchronies (SOAs; 30, 40, 50, 60, 70, 80, 90, 100, 110, and 120 msec) and two separations (10 degrees and 40 degrees) were tested. Perceived velocity estimates were related to the total duration (burst duration + SOA) of the stimulus sequence. The effect of separation on velocity was extremely small but statistically significant. These results are similar to those obtained previously on the minimum conditions for AAM. Duration estimates were related only to total duration, but separation estimates were related to both separation and total duration. These results suggest that velocity is possibly a primary dimension of AAM that is independent of source separation. PMID- 9865084 TI - Inhibition of return to color: a replication and nonextension of Law, Pratt, and Abrams (1995) PMID- 9865086 TI - [Fibrinolysis in the therapy of cerebrovascular accident]. AB - Thrombolytic therapy for acute ischemic stroke based on data reported from recent prospective clinical trials was discussed and analyzed. The author presents some conclusions and suggestions for effective stroke management therapy with rtPA, which he believes will be the future of medical treatment of ischemic stroke. PMID- 9865087 TI - [Role of the implantable cardioverter-defibrillator in secondary prevention of sudden death in hypertrophic cardiomyopathy patients]. PMID- 9865088 TI - [Direct coronary angioplasty versus thrombolysis in the acute phase of myocardial infarction--inpatient outcome]. AB - Both thrombolysis and percutaneous transluminal coronary angioplasty (PTCA) are effective methods for the treatment of acute myocardial infarction (AMI). In our centre we perform primary PTCA during the available schedule of the hemodynamics laboratory. In this article we compare the predischarge evolution of patients submitted to each therapeutic procedure. From January 1996 to June 1997, 298 patients were admitted with the diagnosis of AMI. Eighty-four patients (28%) were thrombolysed (TB group) and 30 patients (10%) underwent primary PTCA (PTCA group). There were no significant differences among the two groups concerning demographic characteristics: age (61 +/- 13--TB and 59 +/- 12 years--PTCA); sex (male 81%--TB; 83%--PTCA), risk factors and previous cardiac history. The mean time since the onset of symptoms until arrival at the hospital was 156 +/- 156 minutes for TB and 202 +/- 210 minutes for PTCA (p < 0.02). The delay since admission until the beginning of treatment was 100 +/- 88 minutes for TB and 119 +/- 142 minutes for PTCA. The primary success rate of PTCA was 94% and there were no complications during the procedure. During the hospital stay, 12 patients developed post-infarction angina in the TB group and two patients in the PTCA group; in 15 patients of the TB group a revascularization procedure was performed (surgery in 5 and PTCA in 10 patients); one patient suffered reinfarction in the TB group. Two patients of the TB group (2.4%) had intracranial hemorrhage; the in hospital mortality was 9.5% in the TB group and 3.3% in the PTCA (p < 0.001). The mean in-hospital stay was 11 +/- 5.6 in the TB group and 7.8 +/- 2.5 days in the PTCA group (p = 0.055). In our experience, primary PTCA in AMI appeared to be a safe procedure with lower occurrence of coronary events and hemorrhagic complication, with an earlier hospital discharge when compared to thrombolysis. PMID- 9865089 TI - [Revascularization in patients with prior coronary bypass surgery]. AB - Increasingly over the past several years, patients have returned after coronary surgery for reintervention procedures. This reflects immediate postsurgical complications and the relentless progression of coronary artery disease in the native circulation and in the bypass grafts. Although there are randomized comparative data for coronary bypass surgery (CABG) versus percutaneous transluminal coronary angioplasty (PTCA) and medical therapy, these trials have always excluded patients with previous (GABG). OBJECTIVES: We attempted to compare the risks and benefits of percutaneous transluminal coronary angioplasty (PTCA) and repeat coronary artery bypass grafting (re-CABG) in patients with previous coronary bypass surgery (CABG). METHODS AND RESULTS: This study examines follow up data (15.4 +/- 11.0 months) from 130 patients with previous CABG, who required either PTCA (Group A, n = 73) or re-CABG (Group B; n = 57) at a single center from 1994 to 1997. Follow up data were obtained from subsequent office visits and telephone calls. The PTCA and re-CABG groups were similar with respect to gender (86% vs 94% males), mean age (62 +/- 9 vs 59 +/- 10 years), angina CCS classes 3 and 4 (73% vs 69%), diminished left ventricular function (23% vs 26%), risk factors such as diabetes (19% vs 17%), hypercolesterolemia (49% vs 45%) and smoking (48% vs 39%) and three-vessel native coronary artery disease (67% vs 72%). The symptomatic status prior to the revascularization procedure was similar in both groups. Complete and functional revascularization was achieved in 85% of the PTCA group and in 92% of those with re-CABG (p = NS). During the hospital stay the complication rates were lower in the PTCA group. Actuarial survival was different at follow up (p = 0.04). Both PTCA and re-CABG groups resulted in equivalent event-free survival (freedom from death, myocardial infarction, unstable angina and urgent revascularization). The need for repeat revascularization at follow up was significantly higher in the PTCA group (PTCA 28% vs re-CABG 10%, p < 0.01). CONCLUSIONS: In this non-randomized study of patients with previous CABG requiring revascularization procedures, PTCA resulted in lower procedural morbidity and mortality risks. At follow up, both PTCA or CABG were similar for event-free survival; PTCA offered lower overall mortality, although it is associated to a greater need for subsequent revascularization procedures. PMID- 9865090 TI - [Myocardial protection against cold stress with pravastatin]. AB - The CARE study showed that the myocardial infarction recurrence rate in patients with moderate cholesterol blood level decreases early during pravastatin treatment. Our goal is to evaluate the possible role of pravastatin in preventing the myocardial lesions induced by cold stress. Twenty Wistar-EPM rats were divided into four groups: Control (CON); PR (Pravastatin) treated with 10 mg/kg/d for 15 days; S (Stress group) in which the rats were submitted to cold stress (-8 degrees C for four hours); and PR + S group treated with pravastatin like PR group and also submitted to the cold stress. The animals were sacrificed and heart fragments were removed for optic and electronic microscopic analysis. The variable considered was mitochondria abnormality (edema, lyses and vacuolization) that was interpreted as crystolyses indices (CI) (n degree of abnormal mitochondria/n degree total of mitochondria). The following crystolyses indices, were found for each group respectively: CON, 2.0%; S, 95.5%; PR, 19.9% and PR + S group, 27.7%*(*p < 0.01). In conclusion, pravastatin prevented myocardial lesions induced by cold stress significantly. PMID- 9865091 TI - [Anomalous origin of the left pulmonary artery (Sling): a case report and review of the literature]. AB - Anomalous left pulmonary artery (vascular sling) is a congenital anomaly in which the vascular structure arises either from the posterior surface of the right pulmonary artery, or from the main pulmonary artery and courses to the left lung between the posterior surface of the trachea and the anterior surface of the esophagus. It may cause compression on the tracheobronchial tree. It is a rare condition leading to death in the first months of life, if it is not corrected. Its diagnosis is quite difficult because it usually presents non specific symptoms frequently associated to diffuse tracheal stenosis. The authors present a clinical case of a newborn with trisomy 21 who had a left pulmonary artery sling associated to tracheal stenosis and congenital heart disease (complete atrioventricular septal defect). They review the literature, particular in what concerns embryopathogenesis, the difficulties in establishing the diagnosis and the surgery which must be performed as soon as possible. PMID- 9865092 TI - [Left ventricular pseudoaneurysm: a rare complication of acute myocardial infarction diagnosed by echocardiography]. AB - Postmyocardial infarction left ventricular pseudoaneurysm resulting from free wall rupture is a very rare finding. Its recognition during life is even rarer. Definitive preoperative diagnosis is difficult, implying, in a great number of cases, the use of multiple imaging modalities. A case of a left ventricular pseudoaneurysm as a mechanical complication of acute myocardial infarction, which was diagnosed by transthoracic echocardiography, is reported. The authors emphasize its rarity, the diagnostic difficulties involved and the contribution of echocardiography to the characterization of that lesion, allowing a definitive diagnosis prior surgery in this case. PMID- 9865093 TI - [State of the art in coronary angioplasty]. AB - Based on a literature overview and on his personal experience with the method, the author analyzes the different phases in the evolution of coronary angioplasty, its indications and limitations in the contemporary setting, as well as the impact of new devices. He emphasizes: the value of the balloon as a basic instrument, followed by the stent; the various attitudes related to the technique; prevention and management of complications during and after the procedure; development, prediction and management of coronary restenosis. PMID- 9865094 TI - [Recommended article of the month: Effects of intensive blood-pressure lowering and low-dose aspirin in patients with hypertension: principal results of the hypertension optimal treatment (HOT) randomised trial]. PMID- 9865095 TI - [Deglutition: physiologic and neurophysiologic aspects]. AB - Swallowing is a complex motor sequence involving the coordinated contraction of many muscles of the buccopharyngeal cavity, the larynx and the oesophagus. Most of the muscles are striated except those of the distal oesophagus which, in human and some other species, are of the smooth type. During swallowing, usually divided into a buccopharyngeal and an oesophageal stage (peristalsis), the sequential activity of the muscles results from motor orders programmed by a rhombencephalic swallowing centre and conveyed to the periphery by efferent fibres belonging to various pairs of cranial nerves (Vth, VIIth, Xth, XIIth). Apart from the motor nuclei of the cranial nerves, the swallowing centre contains an interneurone network responsible for the programming of deglutition. During swallowing, these interneurones (INs) exhibit a sequential activity quite parallel to that of muscles, and persisting in the absence of sensory feedback. The "swallowing INs" are located in two medullary regions: (1) a dorsal region including the nucleus of the solitary tract and the adjacent reticular formation, (2) a ventral region corresponding to the reticular formation surrounding the nucleus ambiguus. The dorsal INs are involved in the initiation and the programming of swallowing. The ventral INs receive their swallowing input from the dorsal neurones and are probably switching neurones that distribute the swallowing excitation to the various pools of motoneurones. The swallowing program can be triggered by inputs originating from either the peripheral reflexogenic areas or the supramedullary structures (cerebral cortex, basal ganglia and hypothalamus). Under physiological circumstances swallowing program is continuously modified by peripheral afferents (expecially muscular) that adjust the force and the timming of contractions to the size of the swallowed bolus. In addition, an important operating feature of the programming network consists of a functionnal polarization so that the activity of proximal portions of the swallowing tract inhibits that of distal portions. This polarization implies the existence of inhibitory connections between interneurones, that could be responsible for the series of delays typical fo swallowing contractile sequence, by generating delayed desinhibitions followed by post inhibitory excitations. Lastly, the sensitive messages that trigger and adjust the swallowing program are at the same time conveyed to higher nervous structures allowing the so called "voluntary" swallow and the integration of swallowing in the ingestive behavior. Disruption of this central loop is likely the source of swallowing disorders (dysphagia) following lesion of cortical or subcortical structures. PMID- 9865097 TI - [Nutritional status and clinical score of 95 patients examined by radiovideoscopy for deglutition]. AB - Ninety five consecutive patients were examined. One performed swallowing videoradioscopy, swallowing clinical score calculation and nutritional assessment with four different methods: Body Mass Index (BMI), arm muscle circumference, dual frequency bioelectrical impedance analysis and serum albumin level. The easiest and the most reliable method for nutritional assessment, comparing to the others, was BMI, where only weight ant height were necessary to measure. So, the prevalence of protein-energy malnutrition was 24.2%, unlinked neither with the presence of videoradioscopy swallowing disorders nor with clinical swallowing score level. The score level was inversely correlated with the presence of videoradioscopy swallowing disorders, and correlated with feeding types: the highest for per os normal feeding, the lowest for patients with parenteral nutrition. The presence of videoradioscopy swallowing disorders was inversely correlated with feeding types. Swallowing clinical score was validated. PMID- 9865096 TI - [Proposal for a predictive clinical scale in dysphagia]. AB - The compiling of a predictive clinical scoring system for the risk of a false passage (FP) has its origins in the limitations of tests which explore only the cough reflex, known to be absent or faulty in 40% of patients investigated for problems of deglutition. A raised clinical score is based on the results of an exhaustive prospective study of 140 patients tested for the first time by videofluoroscopy for FP, this being the reference investigation for false passages. Discriminant analysis allowed the most selective variables to be identified and retained. We have identified 2 categories of patients, according to whether or not they have had any ENT past history. The score is obtained by adding the weighted values for the selected clinical signs. For those patients with an ENT past history, 5 clinical variables were retained: the absence of adhesions = 14, the presence of a velar reflex = 8, the capacity for voluntary deglutition = 8, glottic obstruction = 6, the absence of primitive reflexes = 6, giving a total score for the variables of between 0 and 42. If the total exceeds 32 or is less than 26, there is no FP; for scores between 32 and 26, videofluoroscopy is required to give more precise evidence of the risk. When this scoring system was applied prospectively to a series of 105 patients, a correct predictive result was obtained in 54 patients (51.4%), an equivocal result in 42 patients (40%), and a false result in 9 patients (8.6%), 3 of which (2.8%) were false negatives. The predictive score for FP allows evaluation of the risks of FP in at-risk patients, and adaptation of the treatment strategy according to the type of disordered physiology expected from the cause; thus videofluoroscopy can then be reserved for cases where treatment fails, so that the precise deglutition problem can be pinpointed. PMID- 9865098 TI - Functional swallowing therapy after treatment for head and neck cancer can outcome be predicted? AB - Possible prognostic factors for swallowing after curative treatment of head and neck cancer were investigated by retrospectively analyzing the records of 82 consecutively treated patients. All of them had an exhaustive assessment of swallowing after the oncological treatment, and all had an adequate follow-up. In general, a major improvement in swallowing quality was obser-ved after rehabilitation, although some patients (9/82) still had major problems. Patients with transport problems have a significantly better prognosis than patients with aspiration. No single parameter was strongly correlated with the outcome of swallowing. A weighted score (presence/absence of pharyngeal reflexes combined with the quality of laryngeal sphincter function) was of satisfactory prognostic value for improvement in the quality of swallowing. PMID- 9865099 TI - Ceptral voice analysis: link with perception and stroboscopy. AB - The cepstrum peak magnitude of a /a:/ becomes reduced when either low or high frequency noise increases. In 18 normal subjects and 68 dysphonic patients, perceptual rating according to the GRBAS--system, cepstrum analysis, videostroboscopic vibration pattern quantification, phonation flow measurement, and multidimensional voice analysis was performed. Factor analysis demonstrates that the cepstrum peak magnitude indeed is sensitive to aperiodicity of vocal fold vibration as well as to insufficient vocal fold closure and excessive turbulent noise escape, and that it may be considered as a relevant acoustic correlate for the G (Grade) parameter of the GRBAS scale. PMID- 9865100 TI - Reliability and clinical relevance of perceptual evaluation of pathological voices. AB - The perceptual GIRBAS scale for deviant voice quality was tested in 5 different institutes on 943 voice patients. Each voice was evaluated separately by 2 professionals. The interrater and intrarater correlation is satisfactory for G, R and B. Experience with the scale significantly improves the interrater agreement. Further, GIRBAS/acoustical correlations were investigated by using the multidimensional voice program (MDVP) of Kay. Principal component analysis allows reduction of 21 MDVP parameters to 5 clinically relevant clusters, with a typical acoustical measurement for each of them. The GIRBAS scale seems to be a valuable instrument for clinical practice. PMID- 9865101 TI - Non linear behavior of vocal fold vibration in an experimental model of asymmetric larynx: role of coupling between the two folds. AB - One of the main factors involved in the laryngeal dynamic's non-linearity is the coupling between the two vocal folds. Evaluation of the different coupling elements requires various investigations. The present work evaluate the non linearity effects in clinical situations such as unilateral laryngeal paralysis using Lyapunov's coefficients. It also demonstrate the coupling effects on excised larynx made experimentally asymmetrical, using a specially designed device to measure vibrations from each vocal fold (opto-reflectometer). PMID- 9865102 TI - [Contemporary postural and segmental modification of forced voice]. AB - The forced voice corresponds with increased peri-laryngeal tension during phonation caused by inappropriate muscle action. It is usually accompanied by a characteristic posture (the face thrust forwards, tension in the neck muscles etc...). This posture may cause dysfunction of the cords themselves, leading to dysphonia, and perhaps requiring laryngeal microsurgery. Our project aimed to put forward and validate a system of objective, non-invasive measurements of the postural features of the forced voice, in order to improve the management of these patients. The study was prospective, comparing a control group with no previous vocal problems with a group of patients with problems of laryngeal dysfunction. We carried out a number of simultaneous measurements of postural and segmental activity during phonation, using in parallel an accelerometer, a method of cine-analysis, and a postural platform. Recordings were made during phonation, but only analysed during the resting phase. It turned out that the speed of displacement of the centre of gravity was the most useful discriminating factor. The mean value was 0.07 (displacement type 0.126) for the control group, and 0.32 (displacement type 0.211) for the patient group. The discrimination between these two groups was significant (p < 0.001, Mann-Whitney test). The reliability of this measurement allows us to envisage the inclusion into the clinical investigation package of measurements of the postural and segmental changes in the forced voice. PMID- 9865103 TI - Quantitative rating of video-laryngostroboscopy: a reliability study. AB - The purpose of this study is to examine the most reliable and valid way for rating and comparing videolaryngostroboscopic recordings with a differentiated protocol. 30 high quality videostroboscopic recordings (2-5 minutes) of all kinds of vocal fold pathology were rated independently by 7 experienced laryngologists/phoniatricians, using a standardized and exhaustive protocol form. Interobserver agreement is high for amplitude, mucosal wave, and type of closure/symmetry, is satisfactory for regularity, consistency and vibration of the lesion (if any), and is rather moderate to low for grade of glottic closure and symmetry. As a general rule, raters trained with the protocol form show higher agreement than raters without such experience. Intraobserver consistency is good for all parameters. PMID- 9865104 TI - [Bilateral laryngeal movement disorder and synkinesia: value of botulism toxin. Apropos of a case]. AB - Several years after a subtotal thyroidectomy complicated by bilateral vocal cord paralysis, the patient presented with progressive dyspnea due to laryngeal synkinesis. The impairement of the ventilation status, in spite of laser arytenoidectomy, followed by contralateral posterior transverse cordotomy, suggested a botulinum toxin injection in the intrinsic adductor laryngeal muscles. The rapid improvement in ventilation without phonatory impairement is discussed in the following report. PMID- 9865105 TI - Teflon injection and thyroplasty: objective and subjective outcomes. AB - Management of unilateral vocal fold paralysis continues to generate controversy. Various techniques, but essentially teflon injection and thyroplasty I have been widely used for medialization. Assessment procedures largely differ from author to author: a few authors report objective superiority of thyroplasty (small series). In our own material (19 teflon injections and 9 thyroplasties), when considering objective functional outcomes and patient satisfaction, we cannot demonstrate a statistically significant superiority of one of the techniques, both of them giving very satisfactory results. When considering the literature, the main problem with teflon is delayed (up to 25 years) occurence of granulomas, which is reported as not unfrequent and to some extent unpredictible. Management of teflon granulomas is difficult. Most major complications of thyroplasty (airway obstruction/prosthesis extrusion) occur quite early after surgery and can better be controlled. PMID- 9865106 TI - [Language comprehension by children with profound congenital deafness after cochlear implant]. AB - The consequences of profound deafness on oral language development in children are drastic and well-known. Modern multichannel cochlear implant (CI) has been proven to enhance speech production skills in prelingually deaf children. Speech production skills, however, are known not to be a reliable reflection of oral language competence as a whole. Language is an acquired common code in a specific group, enabling exchange of ideas, feelings and knowledge. In humans, speech is one of the channels conveying language. Assessing language development in CI children is more difficult than simply assessing speech production skills. Many factors may contribute to a poor or an excellent outcome, making it difficult to compare groups of children wearing or not wearing CI. The present study compared receptive language levels in paired matched children from CI and non-CI groups. The main conclusion of this study is that language comprehension scores grow significantly higher over time post-surgery in CI than in paired-matched non-CI children, despite better initial pure tone audiometric thresholds of the latter. PMID- 9865107 TI - [Development of different linguistic systems in children with a cochlear implant]. AB - The authors have studied a population of 21 pre-lingually deaf children equipped with a cochlear implant for at least 24 months. Their linguistic progression after implantation has been compared with the standard language development in normally hearing children, using a model published by Marie Therese Le Normand in 1996. 3 types of development can be distinguished: Profile 1 (shown by 9 children) in which language development after implantation is rapid and homogeneous: profile 2 (shown by 8 children), in which it is slower and more heterogeneous, with a longer pre-linguistic phase; Profile 3 (shown by 3 children) in which there are diverse problems with development. The authors then searched to establish the influence of the age at implantation and the mode of communication on this development. PMID- 9865108 TI - [Re-evaluation of conventional radiography in arthritis]. PMID- 9865109 TI - [Modern imagery in arthritis]. PMID- 9865110 TI - [Role of interleukin-1 in arthritis]. PMID- 9865111 TI - [Production of IL-1 and IL-1 RA in different stages of arthritis]. PMID- 9865112 TI - [Evaluation of the effects of diacerhein in the sheep model of arthritis]. PMID- 9865113 TI - [Efficacy and tolerance of diacerhein in the treatment of gonarthrosis and coxarthrosis]. PMID- 9865114 TI - [Internal meniscal lesions in the stable knee]. PMID- 9865115 TI - [Meniscectomy in arthritis of the knee]. PMID- 9865116 TI - [Follow-up of total knee replacement]. PMID- 9865117 TI - [Controversies in arthritis. I. Arthritis imagery: thoughts on associated pathologies]. PMID- 9865118 TI - [Controversies in arthritis. II. Gonarthrosis: Which surgical treatment to choose?]. PMID- 9865119 TI - [Uveal melanomas and their treatment]. PMID- 9865120 TI - [Transitory visual loss, amaurosis fugax : diagnostic and therapeutic approach]. PMID- 9865121 TI - [Diabetic retinopathy. When should patients be referred to ophthalmologists for evaluation?]. PMID- 9865122 TI - [Modern anti-glaucoma drugs and their secondary effects]. PMID- 9865123 TI - [Diagnosis and treatment of ocular viral infections in AIDS patients]. AB - Ocular complication of AIDS are seen in about 75% of patients. Viral infections are predominant and can involve either external segment in the eye (Herpes type 8 in Kaposi sarcoma, molluscum contagiosum, Herpes simplex and zoster), or the posterior segment of the eye (CMV retinitis). The introduction of a Highly Active Antiretroviral Therapy (HAART) which associates two reverse transcriptase inhibitors and one antiprotease has changed the evolution of AIDS. The decrease of onset of CMV retinitis in AIDS patient is one of the best exemple. For the first time it was possible to stop the maintenance therapy against CMV retinitis in patients that have a sufficient increase in CD4+ cells and they did not present any relapse of CMV retinitis. But an increase of ocular inflammation can be observed with the onset of HAART such as uveitis or cystoid macular edema. PMID- 9865124 TI - [Ocular and palpebral surgery: indications and limits]. AB - Oculo-plastic surgery includes all the problems bound to the eyelids. In this presentation, we expose the most frequent oculo-palpebral pathologies, their treatment and some of the commonest complications. In all facial surgery, there is a plastic component, so it seems impossible to talk about this surgery without taking into account the cosmetical aspects. PMID- 9865125 TI - [Refractive surgery: current indications]. PMID- 9865126 TI - [New frontiers in ocular genetics]. PMID- 9865127 TI - [Elderly patients during an ambulatory neurological consultation]. PMID- 9865128 TI - [Self-administered health questionnaire in the 8th school year]. PMID- 9865129 TI - [Diagnosis and treatment of onychomycoses with terbinafine (Lamisil)]. PMID- 9865130 TI - [Small patients--big costs: the economic aspects of treating young children with renal failure]. AB - Since 1985, 20 children have been followed with early onset of chronic renal failure (plasma creatinine > 120 mumol/l in first year of life). So far, 10 and 7 patients underwent peritoneal dialysis and renal transplantation, respectively. The aim of this study was to assess the overall costs. The recorded costs comprised both the direct costs of dialysis and transplantation, and the costs of all medical and psychosocial measures. The annual median costs of conservative treatment, peritoneal dialysis, the year of transplantation, and follow-up after transplantation amounted to 30,000, 93,000, 130,000 and 28,000 Swiss francs, respectively. The youngest patients caused the highest expenses. The active treatment permitted not only survival, but--in most patients--also a normal cognitive and psychosocial development. PMID- 9865131 TI - [Caustic burns of the esophagus in childhood]. AB - Acute burns of the upper gastrointestinal tract require an immediate diagnostic and therapeutic approach. Early endoscopy has been shown to assist greatly in determining the further treatment of corrosive injury. The main task of radiology is to detect perforations. Early corticosteroid treatment seems to be effective in preventing stricture development in second degree burns. PMID- 9865132 TI - [15 years CA-125 antigen: usefulness and limits. Apropos of 3 clinical cases]. AB - Fifteen years ago, the first clinical assay of the serum tumour marker CA 125 was introduced as a commercial kit. This test is currently the most widely used tumour marker in gynaecologic oncology. Studies have shown that the CA 125 is a valuable tool in the follow-up management of ovarian cancer. As a teaching exercise we present the analysis of three clinical cases, in which CA 125 was determined. Two of these patients did not have an ovarian cancer. It appears that this tumour marker very often is not only used in the follow-up of ovarian cancer, but also as a screening test and as a diagnostic tool. This inappropriate use (due to the lack of specificity and therefore many false positive results) can be avoided by considering it's limits. PMID- 9865133 TI - [Difficulties in differentiation between Crohn disease and ulcerative colitis]. AB - We report on the medical history of a 44 year old woman, who has been diagnosed to suffer from ulcerative colitis, first diagnosed in 1991. Until 1994, when she had finally a colectomy, recurrent episodes occurred under full medical treatment. After colectomy the diagnosis had to be revised as histology revealed Crohn's disease. Only a few months after surgery a new episode of Crohns disease developed including the occurrence of an anal fistula. PMID- 9865135 TI - [General practice case. Hepatitis E with acute jaundice]. PMID- 9865134 TI - [Hypotension in acyclovir therapy]. AB - A 51 year old man developed Herpes zoster on the right arm (C5/C6) treated subsequently with aciclovir infusions (500 mg, 3/day). Ten months before hospital admission he did have a radical resection of a epi-oro-hypopharyngeal carcinoma (T4/N1/G2, M0; lymphangiosis carcinomatosa) as well as a partial laryngeal resection for a recurrence 3 months later and removal of a cervical lymph node metastasis after two further months. During aciclovir treatment the patient experienced repeated bradycardia with hypotension verifiable with the tilt-table test. The bradycardias could not be further characterized by ECC. Neither sonography nor CT-scans gave an indication for infiltration of the cervical course of the vagus or glossopharyngeal nerves. Serum catecholamines were, however, markedly reduced. After cessation of aciclovir the bradycardias and hypotensive episodes disappeared. A final tilt-table test was unremarkable. A reversible autonomic neuropathy induced by aciclovir seems a possible explanation. PMID- 9865136 TI - [General practice case. Erythema nodosum of uncertain etiology]. PMID- 9865137 TI - What the philosophy of medicine is. PMID- 9865138 TI - An analysis of "dignity". AB - The word "dignity" is frequently used both in clinical and philosophical discourse when referring to and describing the ideal conditions of the patient's treatment, particularly the dying patient. An exploration of the variety of meanings associated with the word dignity will note dignity's ambiguous usage and reveal instrumental concepts needed to better understand the discourse of the dying. When applied to a critique of recent and contemporary criticisms of the medical community's handling of the dying, such concepts might provide a more coherent notion of dignity. Rather than a separate construct, a death with dignity might be viewed as an interactive process among the dying and their caretakers. Together, this interdependent amalgam engages in humanizing communication aimed toward understanding the final needs and wants of the patient. PMID- 9865139 TI - Whose consent is it anyway? A poststructuralist framing of the person in medical decision-making. AB - This paper explores the value of a Poststructuralist psychoanalytic model of persons, or "Subjects," as an expanded frame for the question "Whose consent is it anyway?" The elaboration of the need for this expanded frame, its tenets and its value form the substance of the paper. This frame incorporates the emotional, linguistic, and socio-cultural dimensions that help restore patients and physicians to their full status as persons from their restricted status, in the current dominant theory and model, as unidimensional, rationalistic, medico legally constructed players; emphasizes their interconnectedness; and, focuses broadly on responsibility as bearing consequences, and not only accountability. This frame does not deny the role and importance of cognition or rationality, it supplements them. It does not supplant rationality, but rather includes it in a view of the person that also includes those other human capacities which are not based on an ideal of pure reason. PMID- 9865140 TI - Ethical considerations in psychotherapeutic systems. AB - In the process of individual psychotherapy, the client and the therapist work together towards clarifying the client's problems, unlocking vicious circles, opening new perspectives and creating a new narrative congruent with the client's experiencing. The real and undeniable situation in individual psychotherapy across different therapeutic systems is that therapists enter the therapeutic encounter equipped with their own vision of humanity and their own particular theory and methods of psychotherapy. Through the differences in power between therapists and clients and the powerful role of language, clients in their dependent position are apt to assimilate the percepts and the ideas of their therapists. Consequently therapists tend to exert a dominant and influential force in their client's lives. This raises a main ethical concern: to what extent do therapists from different therapeutic systems really help clients to recover their freedom to live their lives congruent to their own authentic perceptions and experiences? PMID- 9865141 TI - A broader look at medical futility. AB - This paper attempts to provide a descriptive theoretical overview of the medical futility debate. I will first argue that quantitative data cannot alone resolve the medical futility debate. I will then examine two aspects of medical futility, which I call the prospective and immediate, respectively. The first involves making prospective factual and value judgments about the efficacy of proposed medical interventions, while the latter involves making value judgments about ongoing medical conditions where the clinical data are clear. At stake is the nature and scope of individual rights. Thus, I maintain there is an undeveloped aspect to the medical futility debate and, briefly, analyze two political perspectives which give rise to different understandings of medical futility. The view that I will defend is that only a system with defined collective goals can accommodate a normative concept of medical futility. These larger questions are the value options which if unaddressed, may be settled by default of economic grounds. PMID- 9865142 TI - [Diagnostic procedure in the woman]. AB - Osteoporosis is characterized by a generalized and progredient bone loss, leading to low bone mass and microarchitectural detoriation with subsequent bone fragility. The same diagnostic approach should be undertaken irrespective of the presence or absence of fractures. The aim of clinical history, physical examination, densitometry and laboratory tests are to exclude diseases that can mimic osteoporosis, to elucidate causes of osteoporosis, to assess severity of osteoporosis and to help selecting the most appropriate form of treatment. PMID- 9865143 TI - [Ultrasound--already clinically relevant?]. AB - The rapid development of Quantitative Ultrasound (QUS) methods in recent years allows to use these devices in a clinical setting. Their primary role is the assessment of fracture risk due to osteoporosis. Further investigations are required for use of QUS for monitoring and diagnostic purposes. Differences between the QUS parameters, Speed of Sound (SOS) and Broadband Ultrasound Attenuation (BUA), between different measurement sites and measurement technology need to be appreciated. Consequently, QUS devices need to be validated individually. In addition, adequate quality assurance measures need to be implemented. QUS and bone densitometry have different complementing features and both should find their place in the clinical arena. These recent developments open up the opportunity to reduce the number of fractures by targeted diagnostic assessments which should lead to medical progress and should also bring along socio-economical advantages. PMID- 9865144 TI - [Biochemical markers of bone metabolism: clinical value in general practice]. AB - Over the past years, the development of new biochemical markers of bone metabolism has generated a substantial amount of data about the pathogenesis, diagnosis, prevention and treatment of osteoporotic bone disease. When used in combination with clinical data and imaging techniques, these parameters permit a comprehensive and non-invasive evaluation of the skeletal status. Biochemical markers of bone metabolism may therefore be used to predict future bone loss and hip fractures in larger cohorts, to select therapy for individual patients, to predict the therapeutic response in individual patients, to monitor therapeutic response and efficacy in, and compliance of individual patients. As far as the initial clinical diagnosis of osteoporosis is concerned, none of the biochemical markers of bone turnover has proven useful as a single determination. Nevertheless, once the diagnosis of osteoporotic bone disease is made, biochemical markers of bone metabolism are useful tools in the further work-up, particularly in identifying patients with high rates of bone turnover. Furthermore, patients with high bone turnover appear to profit from anti resorptive treatments more than subjects with low or normal bone metabolism. The measurement of bone turnover may therefore not only be useful in therapeutic decision making but can also help to reduce costs by identifying those individuals who may not respond well to treatment. Finally, bone markers are of clinical relevance in monitoring both the disease itself and therapeutic efficacy. PMID- 9865145 TI - [Differential osteoporosis diagnosis in the woman]. AB - Bone mineral density of a woman in the second half of her life depends on the amount of bone made during growth and its subsequent rate of loss. Although the rate of bone loss did receive more attention in the study of pathogenesis of osteoporosis, it is becoming increasingly clear that insufficient accumulation of skeletal mass by young adulthood predisposes a person to low bone mass and subsequently to fractures later in life as age related and menopause-related bone loss ensue. In this article we 1) explain the role of inadequate peak bone mass as a major risk factor for osteoporosis and 2) give an overview of factors leading to osteoporosis by decreasing bone mass. Special emphasis has been put on iatrogenic osteoporosis which is frequently neglected because of the fact that the responsible agents often are not known as to be deleterious to the skeleton: among others, glucocorticoids, thyroid hormones and antiepileptics adversely affect bone. PMID- 9865146 TI - [Drug treatment of osteoporosis (in the woman)]. AB - In women, osteoporosis with fractures can happen between 50 years, the age of the menopause, and 80 years. Over these three decades, the condition of bone metabolism is not the same. The years after menopause present with high bone turnover. During these years, the administration of antiresorptive drugs is promising: Estrogens (plus progestagens), calcitonins, bisphosphonates. About 10 years after the menopause bone turnover slows down to low turnover. Now the administration of bone-formation stimulating agents is to be preferred: fluorides in the first line, perhaps anabolics in single cases. The prominent fracture of women older than 75 years is the hip fracture (type II osteoporosis). Bone turnover mostly is accelerated again, now due to secondary hyperparathyroidism as a consequence of long-lasting calcium and vitamin D deficiency. For treatment, calcium plus vitamin D is recommended. The exact diagnosis of the metabolic status of the skeleton is a prerequisite for the choice of the optimal therapeutic principle. PMID- 9865147 TI - [Prevention of osteoporosis]. AB - The European Parliament presented June 10th in Brussels the 'Osteoporosis Report in EU--Means for Prevention'. It was emphasized that in the EU more than 3500 million Ecu have to be spent for hospitalization and that more than 500,000 hospitals beds are being used by osteoporotic patients. According to some calculations this number will double within the next 50 years. The EU has set up eight steps to be considered, e.g. have densitometric measurements available for persons with high risk and have these measurement paid by the insurances to further finance and support research for the very important areas of prevention and treatment. One distinguishes between primary, secondary and tertiary prevention of osteoporosis. Primary prevention aims at reaching at adolescent age a peak bone mass as high as possible. Secondary prevention aims at reducing bone loss peri- and postmenopausal. The tertiary prevention with manifest osteoporosis aims at preventing fractures. Emphasis of the primary prevention is, besides a sufficient calcium intake, to omit risk factors; with secondary prevention the use of medical treatments such as estrogens/gestagens, bisphosphonates, and recently also SERMs is applied. The tertiary prevention tries mostly to reduce the femur fractures. In addition to drugs such as vitamin D/calcium, vitamin D metabolites and bisphosphonates it is very important to create 'a fall-proof home'. Also very useful are hip protectors. PMID- 9865148 TI - [Osteoporosis and nutrition]. AB - Although nutrition accounts only for a small percentage of bone mass, it is essential for reaching and maintaining peak bone mass and for preventing accelerated bone loss with age. Calcium, proteins, and vitamin D are the best investigated nutrients, protein- and vitamin D-deficiency becoming frequent with advancing age, vitamin K, trace elements, and phosphorus having additional effects. Excessive intake of animal proteins leads to bone loss via acidotic stimulation of osteoclastic bone resorption, and high salt intake enhances negative calcium balance by increasing renal calcium losses. Dairy products offer an equilibrated Calcium and protein intake, but have to be monitored for their fat content. Adequate nutrition in childhood influences positively bone mass in adulthood, while nutritional supplementation in elderlies can even increase bone mass. PMID- 9865149 TI - [Osteoporosis in the man: diagnosis and therapy]. AB - The prevalence of osteoporosis in men has been underestimated in the past. Vertebral fractures were found in about 10% in men of age 50 and over 20 to 30% of all hip fractures in advanced age occur in men. Lower life expectancy of men but also differences in bone geometry and remodeling contribute to the lower rate of fractures in comparison to the female gender. In men with suspected osteoporosis a thorough history, physical and clinical examination is mandatory to exclude other localized or generalized osteopathies and to differentiate in primary and secondary osteoporosis. Only some small studies have been published so far on treatment of osteoporosis in men, i.e. therapeutic decisions are mainly based on existing results in postmenopausal osteoporosis. The basis of treatment is calcium and vitamin D supplementation and individually adapted recommendations on life style and risk factor avoidance. In established osteoporosis in adequate analgesic therapy is very important. In cases with secondary osteoporosis, if possible, etiological therapy should be started. Antiresorptive therapy (e.g. calcitonin, bisphosphonates) or osteoanabolic therapy (e.g. fluoride) can later be added, while in idiopathic osteoporosis this is the first option. According to the existing experiences there is in general a good chance to ameliorate the condition in men. There is however an urgent need for further data on therapy of osteoporosis in men. PMID- 9865150 TI - [Physical therapy in prevention and treatment of osteoporosis]. AB - Osteoporotic fractures, and in particular, hip fractures result in significant morbidity and mortality. Low bone mass is the main risk factor of enhanced bone fragility, resulting in an increased risk for hip fracture. Bone density of osteoporotic women with and without hip fractures show a considerable overlap. Therefore, other bone-independent factors also play an important role for the development of hip- and other osteoporotic fractures. One other important factor is falling. In 90% of hip fractures falling was involved [10-15], but only 5% or less of these falls resulted in a subsequent fracture. The view that adequate exercise is beneficial for skeletal health of children and for prevention and treatment of osteoporosis in adults is supported primarily by two lines of evidence: longitudinal and cross-sectional trials in children and young adult athletes showing a significant increase of muscle- and bone mass after strenuous (children) or chronic exercise (athletes) as compared to normally active (children) or sedentary control subjects. What are the potential benefits and limits of specific exercise programs with respect to bone mass, prevention of falls and fractures? In this review these questions are discussed and a specific exercise program in osteoporotic patients with fractures is delineated. PMID- 9865151 TI - Immunological analysis of chromatin: FIS and CHIPS. PMID- 9865152 TI - LIM protein interactions: Drosophila enters the stage. PMID- 9865153 TI - Genomics, Chi sites and codons: 'islands of preferred DNA pairing' are oceans of ORFs. PMID- 9865154 TI - A tale of dwarfs and drugs: brassinosteroids to the rescue. AB - Brassinosteroids (BRs), a specific class of low abundance plant steroids, are capable of eliciting strong growth responses and a variety of physiological changes through exogenous application to plants. Recently, BRs gained general acceptance as important regulators (hormones) of plant growth and development through genetic and molecular identification and characterization of genes involved in BR biosynthesis or response in Arabidopsis, pea and tomato. This major advance in the molecular genetics of BR biosynthesis and mode of action disclosed another case of amazing functional conservation of signalling molecules utilized in plants and animals, and provided a set of extremely valuable tools for the functional analysis of BRs. PMID- 9865155 TI - Counting on comparative maps. AB - Comparative maps record the history of chromosome rearrangements that have occurred during the evolution of plants and animals. Effective use of these maps in genetic and evolutionary studies relies on quantitative analyses of the patterns of segment conservation. We review the analytical methods that have been developed for characterizing these maps and evaluate their application to existing comparative maps mainly for plants and animals. PMID- 9865156 TI - Fragile sites still breaking. AB - Rare fragile sites on chromosomes are the archetypal dynamic mutations. They involve large expansions of the microsatellite CCG or AT-rich minisatellites. The mutation process is an increase in repeat-unit number from within a normal range, through a premutation range, up to full mutation where the fragile site is expressed. Full mutations can inactivate genes and are regions of genomic instability. Common fragile sites, in particular, might have a role in oncogensis by facilitating gene inactivation through chromosomal deletion or amplification, but this requires further exploration. The mechanisms behind the changes that give rise to the cytogenetic manifestation of chromosomal fragility are now beginning to be understood. PMID- 9865157 TI - C. elegans neuroscience: genetics to genome. AB - From their earliest experiments, researchers using Caenorhabditis elegans have been interested in the role of genes in the development and function of the nervous system. As the C. elegans Genome Project completes the genomic sequence, we review the accomplishments of these researchers and the impact that the Genome Project has bad on their research. We also speculate on future directions in this research that are enabled by the efforts of the Genome Project. PMID- 9865160 TI - [Injuries of the extensor tendons: anatomy]. PMID- 9865158 TI - Ageing and mammalian mitochondrial genetics. AB - Mitochondrial DNA (mtDNA) is essential for the ability of mammalian cells to generate a functional oxidative phosphorylation system. Mutations in mtDNA occur in human disease and also during ageing. Here, we address three questions concerning the occurrence and accumulation of mtDNA mutations during the lifespan of the mammalian cell. What sort of mutations accumulate with age in humans and other mammals? How is the female germ line spared from the accumulation of such mutations as occurs in many somatic tissues, so that neonates normally start life with a 'clean sheet'? Is the occurrence of mtDNA mutations associated with the functional decline of cells and tissues during ageing? We argue that mtDNA mutations in somatic cells do not just reflect a passive imprint of ageing, but they are causally associated with the loss of bioenergetic function during the ageing process. PMID- 9865161 TI - [Management and after-care of extensor tendon injuries of the hand]. AB - The investigation and management of extensor tendons play a minor role compared to those of flexor tendons. The finger extension does not seem to be as important as the flexion. But the practical value of the hand is determined by both. Different managements are used depending on the level of extensor tendon injury. More distal located injuries require a longer restraint than those, which are located more proximal. On the one hand this depends on the blood supply of the extensor tendon, which is by far better in the lower arm and dorsal hand than peripheral. On the other hand the reason is the different amplitude of gliding of extensor tendons, which decreases from proximal to distal to lower than 1 mm. Therefore the tension on extensor tendon sutures increases from proximal to distal. The varies techniques of extensor tendon reconstruction will be described. PMID- 9865162 TI - [Facts regarding the Eschede catastrophe]. AB - On 3 June 1998 a high-speed train (Inter City Express, ICE) collided with a bridge at a speed of 200 km/h. The bridge collapsed and the rear wagons of the train were pushed into it with the power of the rear engine. The accident caused 101 deaths and 103 injuries. Four minutes after the accident the alarm was raised, and 16 minutes after the accident the first doctor was on the scene, arriving from Cells, approx. 20 km away. In the first 4 hours after the crash 1844 people from different organisations were at the site of the accident; 461 of these were ambulance personnel and paramedics. 39 aircraft (incl. helicopters and army aircraft) were available at the scene. Many passengers with multiple injuries were stuck in the train and had to be rescued from the severely damaged wagons. Nevertheless all patients with one exception were on the way to hospitals or in the trauma centers 2 hours after accident. 95 passengers passed away on site. The casualties were distributed among 22 hospitals, 2 of them had to be transferred to other hospitals later on for medical reasons. This speaks for the correct selection of hospitals. There were many reasons for the quick rescue and the success of Eschede, namely good weather conditions, the time of the accident (normal working day) the availability of rescue by air, the great number of doctors, ambulance personnel, paramedics and search and rescue teams on site, as well as the excellent cooperation between the different organisations, mainly fire brigades, police, army, border patrol, technical assistance teams and search and rescue coordinators). PMID- 9865163 TI - [Prostheses of the head of the radius. What outcome can be expected?]. AB - We report our results after primary implantation of 30 radial head prostheses in a retrospective study from 1978 to 1996. If there was intraoperatively a remaining instability of the elbow after resection of the radial head, we implanted a prosthesis. The average age of our patients was 40.7 years. The results were evaluated concerning the recommendations of Radin and Riseborough. In 22 patients (73%) we found good to satisfactory results, in 8 patients (27%), there was a free range of motion. Heterotopic ossifications with a remarkable decrease in the range of motion were discovered in 3 patients (10%). The implant had to be removed in three (10%) patients (2 incorrect implantations, one broken prosthesis). To conclude, the primary implantation of a radial head prosthesis after a strict indication improves the clinical benefit for the patient. A correct surgical technique avoids unnecessary complications. PMID- 9865164 TI - [Comparison of a radiation-independent aiming system and free-hand technique for distal locking of unslotted femoral nails]. AB - Although the free-hand technique remains the most popular method for distal interlocking screw insertion, proximally mounted "radiation-independent" devices which compensate for implant deformation recently have been developed for rotationally stiff, unslotted femoral nails. However, the benefits of such systems have not been determined. This study prospectively compared the duration of the nailing procedure, the length of radiation time, and the accuracy of interlocking screw placement with a radiation-independent distal aiming system to those using the free-hand technique. In twenty paired intact cadaveric femora, a single surgeon experienced only in the free-hand technique performed statically locked intramedullary nailing using the two methods. For the aiming system and free-hand technique respectively, the total operation time was 19.1 +/- 8.4 versus 20.9 +/- 11.3 minutes (p = 0.9), the distal locking time was 6.6 +/- 2.4 versus 4.8 +/- 1.5 minutes (p = 0.002), the total fluoroscopy time was 23 +/- 17 versus 69 +/- 34 seconds (p < 0.0001), and the distal locking fluoroscopy time was 0 versus 37 +/- 15.5 seconds (p < 0.0001). There were no failures in either group. Drill nail contact (p < 0.0001) and distal screw damage (p = 0.02) were both greater with the free-hand technique. PMID- 9865165 TI - [Clinical results of primary intramedullary osteosynthesis with the unreamed AO/ASIF tibial intramedullary nail of open tibial shaft fractures]. AB - From 04/91 to 06/96 sixty-nine open fractures of the tibia were primarily treated on the day of the accident with unreamed nailing (UTN, Synthese). The distributions of fracture type according to the AO classification and of soft tissue injury according to Gustilo were as follows: fracture type: A: 28%, B: 52%, C: 20%; soft tissue injury: I: 30%, II: 28%, IIIA: 12%, IIIB: 12%, IIIC: 6%. Of the 65 fractures assessed 46 (71%) healed within 18 weeks without secondary intervention. There was delayed healing in three fractures requiring secondary conversion to reamed nailing. Eight fractures (12%) developed pseudarthrosis of which five (8%) healed uneventfully. Deep infections was manifest in four fractures (6%). Three of these infections developed after secondary intervention to treat pseudarthrosis. Seven of the eight pseudarthroses and three of the four infections healed eventually. Revision procedures were necessary in 11 patients (17%) to deal with disturbed fracture healing or infection (10 reamed nailing procedures, three cancellous bone grafts, and one of each of the following: sequestrectomy, fibular osteotomy, plate fixation, external fixator, monorail procedure). The results show that the same good infection rates were achieved for the UTN as for the external fixator. The advantages of the UTN are, however, a lesser need for secondary intervention and greater patient comfort. Therefore, we find the UTN to be a good alternative to the external fixator in the treatment of open fractures with severe soft tissue damage. PMID- 9865166 TI - [Quality of life after complex tibial reconstruction]. AB - 58 patients following Gustilo IIIB open tibial fractures, treated by serial debridements, osteosynthesis and microvascular free tissue transfer were investigated after an average of 6 years. Assessment of life satisfaction was done by means of a proven valid and reliable test instrument (FLZ-"Fragen zur Lebenszufriedenheit"). Further specific questions on treatment satisfaction, aesthetic result, occupation and income were added to this questionnaire. Significant discrepancies of the items recreational activities, income, occupation and partnership, productivity, mobility and pain were evaluated in comparison to available data of healthy west german population. The calculated global index of life satisfaction and health differed significantly from the established index of west german population. Although specific aspects like aesthetic result, donor site morbidity, pain and even function were critically judged, 91% of the patients were satisfied with the long term result of the treatment. None of the patients thought that an initial amputation probably might have been a better solution. The postrehabilitation employment rate was 64%. Limb salvage of Gustilo IIIB fractures by means of a multidisciplinary treatment protocol is able to afford useful and satisfying long time results. However, an impairment of quality of life, at least in specific items has to be considered. PMID- 9865167 TI - [Trendy inline skating sports. Pattern of injuries and groups at risk]. AB - One hundred and eighty-two patients presented with 200 inline-skating injuries over a 30-month period. Of these, 14% were admitted to hospital, 10% required operative treatment. Fractures (49%), contusions/lacerations (27%) and capsular/ligamentuous injuries (16%) were the most prevalent types of injury. Children had an even higher risk of sustaining fractures (62%). Falling on the extended arm caused 44% of all injuries. Thirteen percent resulted from torque mechanisms of the leg, whereas direct trauma to elbow or knee (5% each) were uncommon injury mechanisms. Injuries of the elbow, forearm, wrist and hand accounted for 55.5% of all cases and 71% of all fractures. Head (13%), knee (9.5%) and ankle (9%) were other regions frequently involved. Protective equipment was often used only for uninjured regions, whereas the injured regions had most often been left unprotected. PMID- 9865168 TI - [Prevention of thrombosis with physical therapy in trauma surgery. Possibilities and value of individual measures]. AB - Patients in traumatology need additional physiotherapy to prevent deep vein thrombosis. There is still a 4% risk of deep vein thrombosis in outpatients receiving already low molecular weight heparin prophylaxis [9]. However, there are no prospective studies with a high number of patients to prove effectiveness of the presented physiotherapeutic tools, even if the hypothesis "achieving physiological-like conditions" sounds positive and some authors postulate that physiotherapy would be equal to drug prophylaxis of deep vein thrombosis. PMID- 9865169 TI - [Complications in trauma surgery]. PMID- 9865170 TI - [Resorbable interference screws. Histologic study 4.5 years postoperative]. AB - The use of degradable implants in recent years has brought not only good results. Foreign body reactions to implantation of degradable polymers clearly curtailed the initial expectations. Bioabsorbable polymers have been used for sutures, vascular clips, and fixation implants for both intra-articular fractures and malleolar fractures. In the literature these implants are controversial. Potential benefits remain for the use of this material for interference screws if the problems of inflammation are minimal and the fixation properties are sufficient. There is no need for a second operation for removal. In one patient, 4,5 years after to implantation of an degradable interference screw (Copolymer 85/15 D,L lactide glycolide; Biologically Quiet, Instrument Makar, Inc. Mount Hope, MI 48864, USA) we took an biopsy and made histological examinations. No foreign body reaction or inflammation signs could be detected. PMID- 9865171 TI - [Using autoclaved spongiosa]. AB - Since transmission of HIV through allogenic bone grafts has been established, the concept of cryopreservation of allogenic bone had to be reconsidered. The strict guidelines of the scientific board of the Bundesarztekammer of 1990 are very labour-, time- and money-intensive. We have therefore moved to autoclaving allogenic cancellous bone. This is harvested from femoral heads during THR in slices of 2 to 3 mm, then cleaned under non-sterile conditions with a hard water jet followed by an ultrasonic bath for approx. 20 minutes. The slices are then double sealed individually and autoclaved at a temperature of 134 degrees C and a pressure of 2.5 atmospheres. Storage is in sealable containers at room temperature, so the material available at any time. Since the bone tissue has been freed of most organic matter and therefore lost its bone-specific antigenic structure, all that is left is the anorganic component with its inimitable architecture. This treatment results in a cancellous bone graft which is sterile, biocompatible and osteoconductive. Biologically it is inferior to autogenic and cryopreserved bone. Experimental and clinical studies show, however, that autoclaved cancellous bone can be a suitable substitute in well-selected indications. To elucidate the ultrastructural changes of the spongiosa and proteins induced by autoclaving further investigations are necessary. For example, the specific proteins involved need to be determined. PMID- 9865172 TI - [The 25th anniversary of the Research Institute of Eye Diseases of the Russian Academy of Medical Sciences]. PMID- 9865173 TI - [Intraocular circulation in glaucoma]. PMID- 9865174 TI - [The surgical treatment of normotensive glaucoma]. AB - The efficacy of surgical treatment of low (normal)-pressure glaucoma is validated on the basis of analysis of the disease progress risk factors. The risk factors are inadequate intraocular pressure (IOP) and insufficient blood supply to the optic nerve and retina. For stabilizing the glaucomatous process, IOP is to be reduced below 14 mm Hg. After antiglaucoma surgery, IOP decreases by 35% on average (to 13.9 mm Hg), and visual field is retained in remote period in 81.2% patients. Antiglaucoma operation with simultaneous decompression of the optic nerve results in a greater increase of visual field due to improvement of blood supply to the optic nerve and retina; visual functions are stabilized for a long time in 77.8% cases at a higher IOP (16.14 mm Hg) in patients with worse initial status. Functional results of surgery for normotensive glaucoma depend on ophthalmic tone and optic nerve and retinal hemodynamics. PMID- 9865175 TI - [The outlook for the use of diode laser radiation for trabeculoplasty in treating primary open-angle glaucoma]. AB - Clinical efficacy of diode laser trabeculoplasty is assessed in 35 eyes of 30 patients with primary open-angle glaucoma during during the initial and well developed stages of diseases. Changes in the anterior segment of the eye, intraocular pressure, ocular hydrodynamics, visual functions, and status of the optic nerve were assessed 6-12 months after the operation. Diode laser trabeculoplasty is characterized by areactive clinical course and leads to a significant decrease of intraocular pressure, stabilizes and improves visual functions, and ameliorates intraocular humor discharge. The treatment runs a smooth course without complications. The method is recommended for clinical practice and further studies. PMID- 9865176 TI - [Clinico-prognostic criteria for the selection of diabetes mellitus patients for the intraocular correction of aphakia]. AB - The level of glycosylated blood hemoglobin (HbA1c) as a clinical and prognostic criterion for selecting diabetics for implantation of intraocular lens (IOL) is assessed. Extracapsular cataract extraction with intraocular correction was carried out in 68 patients with noninsulin-dependent diabetes mellitus. Preoperative level of HbA1c in group 1 (n = 36) was no more than 8%, and in group 2 (n = 32) it was at least 8%. Both groups were treated similarly. The immediate and late functional results of cataract extraction with IOL implantation were decisively better in group 1 with HbA1c level under 8%, that is, with compensated diabetes. Thus, the level of HbA1c is an accurate and objective clinical and prognostic criterion for selecting patients with diabetes for IOL implantation with good and stable functional results and for precluding unjustified operations. PMID- 9865177 TI - [Hermetic sealing of the cataract incision with intracorneal mattress sutures]. AB - Main factors of hermetic closure of an operative wound in cataract extraction with intracapsular implantation of intraocular lens are discussed. A new method for closure of operation wound in this operation is proposed. Opposition intracorneal mattress sutures are made over the incision plane. Their principal feature is that the thread is not thrown over the external edge of corneal incision, as in traditional non-opposition sutures, and during pulling and suture the thread does not deform the external surface of the cornea in the central zone and hence, causes no postoperative corneal astigmatism. A method combining opposition and interrupted non-opposition sutures is proposed, and various combinations of these sutures are compared. Forty-five patients (45 eyes) were operated on using the proposed technique. Opposition sutures made in accordance with the methods proposed by the authors do not cause high postoperative corneal astigmatism, or it approximates the normal values of corneal astigmatism. A special marker is proposed for preliminary marking of sites for nonperforating incisions on the cornea and the site of cataract incision. PMID- 9865178 TI - [Objective indices of corneal transparency and of fluorescein permeability through the cornea before and after photorefractive keratectomy]. AB - Video images of the cornea in photorefraction keratectomy (PFK) are analyzed by computer-aided microdensitometric analysis of televideo images of the eye. Optic density of various layers of the cornea and fluorescein penetration through the cornea were assessed in patients with myopia (235 eyes) and hypermetropia (65 eyes) before the after PFK. PFK was carried out by Nidek excimer laser (Japan). Analysis was carried out with consideration for the degree of emmetropia and patients' ages. Corneal transparency and index of fluorescein penetration through the cornea correlated with patient's age and emmetropia. Computer-aided microdensitometric analysis of televideo images of the eye earlier detects changes in the cornea than routine methods. The authors discuss the probability of assessing corneal transparency for selection of patients for PFK. PMID- 9865179 TI - [The efficacy of using enzymes in treating toxic-allergic reactions after cataract extraction with intraocular lens implantation]. AB - Physiotherapy was administered to 84 patients aged 59-78 years with toxic allergic reaction in the eye after extracapsular cataract extraction with implantation of intraocular lenses (IOL). The patients were divided into 4 groups. Group 1 (n = 15) were treated by fibrinolysin magnetophoresis, group 2 (n = 17) lidase magnetophoresis, group 3 (n = 28) lecozyme magnetophoresis, and group 4 (n = 24) collalysin magnetophoresis. Treatment started on days 3-5 after surgery. Magnetophoresis was carried out using Russian Gradient-1 device, generating a low-frequency magnetic field of 10-12 mT, 10-12 min exposure. For patients with pronounced iridocyclitis and uveitis, enzyme magnetophoresis was supplemented with atropine, adrenalin, and calcium chloride electrophoresis (with monitoring the pupil size). Comprehensive treatment resulted in complete resolution of exudation in the anterior and posterior segments of the eye, hypopyon, loosening and resorption of anterior and posterior synechiae, the vitreous body grew more transparent, visual acuity improved in 91.1% patients (coarse dystrophic changes in the central zone of the fundus oculi were detected in 8.9%). Magnetophoresis with fibrinolysin and lidase is most effective in patients with toxic allergic reaction during the early periods after cataract extraction with IOL implantation, while in later periods magnetophoresis with lecozyme and collalysin should be preferred. The efficacy of collalysin is not inferior to that of lecozyme, but its effect is more mild and the drug causes no allergic reactions. Magnetophoresis with mydriatics (with monitoring the pupil size) and calcium chloride is recommended for all cases. PMID- 9865181 TI - [Intubation granulomas of the lacrimal ducts in patients with silicone implants]. AB - The results of 315 endonasal operations on the lacrimal duct with long intubation are analyzed. The most incident complication of long intubation of the lacrimal duct is formation of intubation granulomas, which are most often localized in the inferior lacrimal canaliculus; their etiology is sometimes allergic--they form as a response of adjacent tissues to the material of the intubation tube, and the findings of our recent histological study prove it. The diagnosis of intubation granulomas is easy in the majority of cases. The treatment consists in removal and obligatory adequate total and local antiallergic therapy decelerating the growth and relapses of granulomas. After removal of the intubation tube the growth of granulations ceases and a fine cicatrix forms. PMID- 9865180 TI - [The clinical characteristics and immunotherapy of complicated forms of adenovirus keratoconjunctivitis]. AB - Analysis of case histories and clinical and direct immunofluorescent examinations of 72 patients enabled the authors to single out 3 forms of complicated adenovirus keratoconjunctivitis (AVKC): 1) acute grave; 2) with toxic allergic reaction to previous treatment; and 3) steroid-complicated. Recommendations on the treatment of these forms are offered. A common remedy is interferon inductor poludan and chigain obtained from human colostrum and containing secretory IgA. Poludan was administered by instillations (4-6 times a day) and periocular injections in a dose of 100 U every 1-2 days, 5-6 injections per course, chigain was administered by instillations (2-4 daily). This combined treatment was highly effective and well tolerated. Periocular injection of poludan ensured much more intensive local and even systemic interferon production than instillations alone. Mean terms of treating complicated AVKC were compatible with those in uncomplicated forms: 14.3 +/- 2.1 vs. 13.2 +/- 1.2 days, respectively. Signs of herpesvirus infection were detected in one-third of patients with steroid complicated AVKC. For eliminating corneal opacities in AVKC patients, enzyme phonophoresis, phototherapeutic keratectomy, and (in grave cases) lamellar keratoplasty are recommended. PMID- 9865182 TI - [The characteristics of the clinical course in individual types of radiation induced cataracts]. AB - Twenty-eight patients exposed to acute and chronic ionizing radiation were followed up. Radiation cataracts were diagnosed in 22 eyes. Localizations of changes in the lens after ionizing and solar radiation are similar. Opacities are localized under the posterior lenticular capsule, round the nucleus, and under the anterior capsule and do not depend on the type of radiation. Opacities caused by solar radiation are notably smaller and look as numerous small dots. Study of cataracts of different origin and assessment of the efficacies of different anticataract drugs were carried out for evaluating the progress of different types of cataracts (cellular, extracellular, and capsular forms). PMID- 9865183 TI - [Viral lesions of the eye in human immunodeficiency virus infection]. AB - HIV infection destroying cellular immunity system creates prerequisites for herpesvirus involvement of the anterior and posterior segments of the eye. A total of 370 HIV-infected patients were examined. Ocular diseases were detected in 53 (14.35%). The most prevalent conditions were retinal angiopathy (most often with cotton-like foci--in 7.3% and cytomegalovirus (CMV) retinitis in 4.1%). Cotton-like foci in the presence of CMV activation (evidenced by high level of CMV DNA in peripheral blood) are a sign of probable CMV infection manifesting by retinitis or involvement of other organs. Intravenous gancyclovir in a dose of 7.5 mg/kg twice a day and its combination with interferon inductor poludan are effective methods for therapy of CMV infection, preventing blindness or poor vision. Moreover, combination of chemotherapy with interferon inductors cured involvement of the anterior segment of the eye--herpetic keratitis and iridocyclitis, which sometimes develop in the presence of HIV infection. PMID- 9865185 TI - [The computer processing of B-echo scanograms of the eye in hemophthalmos]. AB - A total of 260 patients with uncomplicated hemophthalmia of different origin (eye contusions, essential hypertension, diabetes mellitus) are examined. A method for quantitative assessment of ultrasonic scanograms of the eye by microdensitometry and computer analysis of total intensity (acoustic density) and area (volume) of hemophthalmia is proposed. PMID- 9865184 TI - [The hemodynamic characteristics of the central retinal artery and ophthalmic artery in atherosclerotic lesions of the carotid arteries based on the data from ultrasonic study methods]. AB - Central retinal artery and orbital artery were examined by ultrasonograph in 348 patients with atherosclerosis of the carotid arteries. Doppler shift of frequency spectrum was detected in the central retinal artery, which is typical of the ischemic syndrome of the eye. Its characteristics include decreased diastolic component of velocity and increased resistance index. The possibility of using duplex scanning with color Doppler flow mapping for assessing the treatment efficacy is demonstrated. A direct relationship between the degree of stenosis in the internal carotid artery and blood flow in the orbital artery is revealed. The detected changes do not notably affect the Doppler spectrum characteristics in the central retinal artery. PMID- 9865186 TI - [The prognostic significance of the electrophysiological indices of the retina and optic nerve in patients with opacities of the optical media of the eye]. AB - Prognostic significance of the main electrophysiological parameters (electroretinogram, electrocoagulogram, visual evoked potentials), and electric sensitivity of the eye in diseases of the retina and optic nerve, and their informative value in patients with opaque optic media of the eye are discussed. The authors note that decrease of the total retinal biopotential and constant potential not always characterize the level of its viability (in retinitis pigmentosa, detachment, vitreous opacities, etc.). Insufficient informative value of visual evoked potentials in patients with opaque optic media is explained by impossibility of presenting pattern stimuli. Many-year clinical experience gained in study of electric sensitivity in more than 40,000 patients with diseases of the neuroreceptor system of the eye brought the authors to a conclusion that this method allows rapid assessment of the total area of visual field and optic nerve conductivity. Study of the threshold electric sensitivity in patients with opaque optic media is in fact the only method for assessing peripheral vision and viability of a detached retina. PMID- 9865187 TI - [The role of the mast cells of the conjunctiva in the intercellular interactions in keratoconus and epithelial-endothelial corneal dystrophy]. AB - The aim of this study was assessment of the role of mast cells in inflammatory processes associated with keratoconus and corneal epithelial-endothelial dystrophy and clinical assessment of Lecrolin, a specially designed dosage form (eye drops) of potassium cromoglycate (intal) used as an antiallergic drug in asthma, rhinitis, etc. Examinations of 43 biopsy specimens of the conjunctiva from patients with keratoconus and epithelial-endothelial corneal dystrophy demonstrated the significance of immune inflammation in the pathogenesis of these diseases. Mast cells are a typical cell element participating in inflammation at all stages. The count of mast cells increases from 2.5 during the acute stage to 8.86 during the chronic stage, which is explained by absence of the leukocytic stage and proteolysis and stabilization of mature labrocyte degranulation associated with it. Reactions of mast cells with lymphocytes and monocytes/macrophages lead to proliferation of fibroblasts and chronic development of collagen formation. Close relationships between microvessels and mast cells promote stable spasms of the vessels, impairing the microcirculatory bed and leading to ischemia of the anterior segment of the eye. In such cases sodium cromolyne drugs (lecrolin) inhibiting histamine release and stabilizing mast cell membranes are recommended. PMID- 9865188 TI - [A pathohistological study of newly formed subpigment epithelial tissue in patients with senile macular dystrophy]. AB - Efficacy and possibility of direct operations on the macular area of the retina in humid (exudative) maculopathies (senile maculodystrophy) is validated theoretically, morphologically, and clinically. A hypothesis on the mechanism of exudative process development in the macular area in degenerative diseases is formulated on the basis of pathohistological analysis of removed subpigmental material. Indications for such operations are defined. PMID- 9865189 TI - [An analysis of the prevalence of clinical refraction (based on data from the reciprocal dispensary care of students at the I. M. Sechenov Moscow Medical Academy)]. AB - Results of mutual check-ups of 5-year students of therapeutic and medical prophylactic faculties of I. M. Sechenov Moscow Medical Academy in 1991-1994 are analyzed. A total of 584 case histories of students are reviewed. The data demonstrated correlation of many signs in the sample and statistical data of ophthalmological publications on refraction. Conclusions about errors in prescription and use of eyeglasses are made. Violation of visual loading regimen by these students are detected. PMID- 9865190 TI - State profiles on women's health. PMID- 9865191 TI - High-risk nonmelanoma skin cancers. PMID- 9865192 TI - Laser scar revision: comparison of CO2 laser vaporization with and without simultaneous pulsed dye laser treatment. AB - BACKGROUND: Over the past decade, the 585-nm pulsed dye laser (PDL) has been used successfully to treat a variety of cutaneous vascular lesions as well as hypertrophic scars. Laser scar revision has been revolutionized by the recent development of high-energy, pulsed carbon dioxide (CO2) laser systems. These new CO2 lasers allow controlled vaporization of thin layers of skin while minimizing damage to surrounding dermal structures. OBJECTIVE: To determine the effect of a high-energy, pulsed CO2 laser alone and in combination with a 585-nm PDL on nonerythematous hypertrophic scars. METHODS: Twenty patients with nonerythematous hypertrophic scars were treated with a high-energy, pulsed CO2 laser. One-half of each scar was additionally treated with the 585-nm PDL laser. Sequential clinical and photographic analyses were performed independently by two blinded assessors. In addition, erythema reflectance spectrometry measurements were obtained from the scars before and at regular postoperative intervals. RESULTS: Global assessment scores and erythema spectrometry measurements were significantly improved after laser treatment. Combination CO2 and PDL laser treatment resulted in more significant improvement than CO2 laser irradiation alone. CONCLUSION: Concomitant use of the high-energy, pulsed CO2 and PDL laser systems was superior to CO2 laser vaporization alone for revision of nonerythematous hypertrophic scars. Once again, the vascular specificity of the 585-nm PDL has been linked to improvement in hypertrophic scar tissue. PMID- 9865193 TI - Erbium:YAG laser resurfacing in Asians. AB - BACKGROUND: Although laser resurfacing has become increasingly popular in the treatment of Caucasian skin, concerns about healing and postinflammatory pigmentary changes have limited its use in Asian skin. OBJECTIVE: The purpose of this study was to determine the efficacy and safety of the Erbium(ER):YAG laser used in the treatment of Asian skin. METHODS: Fifty Asian individuals with Rhytids, scars, pigmentary alteration, and a variety of cutaneous growth were treated with the Erbium:YAG laser. Patients were evaluated for clinical efficacy, wound healing, post-operative erythema, and post-inflammatory pigmentary changes. RESULTS: Significant improvement was noted in all individuals. Shorter periods for re-epithelization and erythema duration were noted when compared to previously reported results following carbon dioxide laser resurfacing. CONCLUSION: The Erbium:YAG laser is safe and effective in the treatment of Asian skin. PMID- 9865194 TI - Successful removal of traumatic tattoos in Asian skin with a Q-switched alexandrite laser. AB - BACKGROUND: Traumatic tattoos result from mechanical penetration of the skin by foreign-body particles associated with puncture, abrasive, or explosive trauma. Until the recent development of the Q-switched lasers, it was not possible to remove tattoo pigments without scar and pigmentary changes. OBJECTIVE: The objective of this study was to determine the effectiveness of the Q-switched alexandrite laser (wavelength, 755 nm; pulsewidth, 100 ns), in treating the 27 cases of Asian skin with 36 traumatic tattoos and to observe any side effects such as scarring or pigmentary change. METHODS: The results of treatments on 16 patients with 19 penetrant tattoos, 10 patients with 16 abrasive tattoos and 1 patient with bomb explosion were clinically analyzed. RESULTS: Greater than 76% removal of tattooed pigments required an average of 1.7 treatment sessions in penetrant tattoos in contrast with 2.4 sessions in abrasive tattoos. The excellent removal of traumatic tattoos required 7.5 J/cm2 except the scarred region of one explosive tattoo and one abrasive tattoo on soil. There were no permanent side effects such as scar or permanent pigmentary changes. CONCLUSION: In conclusion, the Q-switched alexandrite laser is a safe and highly effective modality for removal of various traumatic tattoos without scar or permanent pigmentary change in Asian skin. PMID- 9865195 TI - Quantitative comparison of inflammatory infiltrate and linear contraction in human skin treated with 90-microsecond pulsed and 900-microsecond dwell time carbon dioxide lasers. AB - BACKGROUND: Skin resurfacing with 90-microsecond pulse duration carbon dioxide (CO2) resurfacing lasers has been reported to have shorter duration of erythema compared with skin resurfacing with 900-microsecond dwell time lasers. The presence of inflammatory infiltrate following resurfacing may correlate with the persistence of this erythema. Furthermore, skin treated with the 90-microsecond pulse duration laser and the 900-microsecond dwell time lasers both result in equivalent improvement of rhytids in the treated skin. OBJECTIVE: To quantitative the inflammatory cell infiltrate and linear contraction of skin treated with the 90-microsecond pulsed and 900-microsecond dwell time CO2 lasers at intervals of 2 and 4 weeks after treatment. MATERIALS AND METHODS: Volunteers were recruited from patients who were planning to undergo full face laser resurfacing under general anesthesia. Informed consent was obtained from all volunteers. In the posterior auricular areas of all volunteers, four separate rectangular areas were marked using a skin marking pen and a template. Two rectangular areas behind the right ear were treated with 6 passes of the 90-microsecond laser and two rectangular areas behind the left ear were treated with the 900-microsecond dwell time laser. The resurfaced areas were wiped with a moist cotton swab and then patted dry with dry gauze between passes. Contraction measurements of the resurfaced areas were taken before and immediately after laser treatment and again at 2 and 4 weeks following treatment. Punch biopsies were also performed at 2 and 4 weeks after treatment in an area of skin different from where contraction measurements were taken. RESULTS: The number of inflammatory cells present in the skin at 2 and 4 weeks after laser resurfacing are greater for skin resurfaced with a 900-microsecond dwell time laser than a 90-microsecond pulse time laser. Linear contraction of skin immediately after treatment was 18% greater with the 900-microsecond dwell time laser than with the 90-microsecond pulsed laser. The difference in the amount of contraction produced by the lasers tended to decrease over time. At 4 weeks there was a 10% difference in mean linear contraction between the two laser types. CONCLUSION: Increased numbers of inflammatory cells in skin resurfaced with the 900-microsecond dwell time laser may explain the observed persistence of erythema associated with the 900-microsecond dwell time laser. Measurable linear contraction produced by the 900-microsecond dwell time laser was initially 18% greater than the 90-microsecond pulse laser. This difference tends to decrease over time. PMID- 9865197 TI - Evaluation of blood loss during tumescent liposuction in Orientals. AB - BACKGROUND: There are high risks from general anesthesia and excessive bleeding associated with traditional liposuction using the dry or wet method. The blood loss has been estimated to be between 15% and 45%. The tumescent technique permits liposuction of more than 3,000 ml of fat totally by local anesthesia without sedation. The blood loss is reported to be less than 1% of aspirate. OBJECTIVE: To examine the blood loss during liposuction using tumescent technique in Chinese patients compared with that of Caucasians as published in the literature. METHODS: Hemoglobin values of patient's preoperative venous blood and infranatant of aspirate were measured in 45 consecutive cases of liposuction using the tumescent technique from May 1996 to June 1997. RESULTS: A total of 30 patients completed the study. The average blood loss was estimated to be 1.08% of aspirate. CONCLUSION: From this study, we found the blood loss in Chinese patients is comparable with that in Caucasians. Tumescent liposuction is a safe dermatologic cosmetic surgical procedure without the need of blood transfusion and general anesthesia. PMID- 9865196 TI - Injectable hyaluronic acid gel for soft tissue augmentation. A clinical and histological study. AB - BACKGROUND: Several biomaterials are available for the purpose of soft tissue augmentation, but none of them has all the properties of the ideal filler material. The recent development of hyaluronic acid gels for dermal implantation give the physician new possibilities of effective treatment in this field. OBJECTIVE: This study provides a clinical and histological evaluation of safety and efficacy of a cross-linked stabilized non-animal hyaluronic acid gel (Restylane, Q-Med, Uppsala, Sweden) to determine its characteristics, advantages, disadvantages, and side-effects. METHODS: 158 patients were treated with facial intradermal implant of hyaluronic acid gel for augmentation therapy of wrinkles and folds, and for lip augmentation and/or recontouring. The results were evaluated in all patients by subjective judgement by the physician and the patient, and by photographic method at time 0 and after 1, 2, 4 and 8 months from the procedure. In addition, a smaller histological study was carried out in five volunteer patients for a term of 52 weeks to determine the interaction and duration of the material in human healthy skin. RESULTS: Clinically, both the physicians' and patients' evaluations revealed very satisfactory results, with a global 78.5% and 73.4% respectively of moderate or marked improvement after eight months, independent of the treated area. The photographic evaluation revealed even better results with a 80.4% of moderate or marked improvement after 8 months. The safety evaluation showed a 12.5% of postoperative immediate adverse events, that were localized and transient. There was no evidence of major systemic side effects. Histologically, the product was shown to be long-lasting and well tolerated as judged by histological techniques. CONCLUSIONS: Stabilized, non-animal, hyaluronic acid gel is well tolerated and effective in augmentation therapy of soft tissues of the face. This material presents several advantages in comparison to previously used injectable biomaterials and expands the arsenal of therapeutic tools in the field of soft tissue augmentation. PMID- 9865198 TI - Prevalence of male pattern hair loss in 18-49 year old men. AB - BACKGROUND: Previous studies investigating the prevalence of male pattern hair loss (MPHL) typically used biased samples of men recruited from clinical populations which may limit generalizability of findings to broader populations. OBJECTIVE: To obtain an updated and improved estimate of the occurrence of MPHL in healthy men residing in the community. METHODS: Community-based sample of healthy men aged 18-49 years participated in a study investigating the effects of MPHL. Participants completed a brief questionnaire self reporting degree of hair loss, general health-related quality of life (HRQL) and hair-loss-specific measures. A trained observer also rated each participant using standardized classification for MPHL. RESULTS: The proportion of men with moderate to extensive hair loss (type III or greater) was 42%. The proportion of men with moderate to extensive hair loss increased with increasing age, ranging from 16% for men 18-29 years of age to 53% of men 40-49. Twelve percent of the men were classified as having predominantly frontal baldness (type A variants). CONCLUSIONS: MPHL, especially frontal baldness, may be more common than previously reported. PMID- 9865199 TI - The use of the SMAS to close Mohs defects invading the parotid gland. AB - BACKGROUND: Plastic surgery literature reports the use of the superficial musculoaponeurotic system (SMAS) in reconstruction after parotidectomies to prevent the complications of Frey's Syndrome and fistula formation. However, there is scant information in the dermatologic literature regarding the use of the SMAS when closing Mohs defects overlying the parotid gland. OBJECTIVE: Our purpose is to introduce to the dermatologic surgery literature the need for plication of the SMAS prior to closure of the overlying tissues when repairing surgical defects which invade the integrity of the parotid gland. METHODS: We describe a patient with a large basal cell carcinoma located in the right preauricular region. Using text and photographs, we detail our closure of this large defect which invaded the parotid gland. RESULTS: Closure of the SMAS over the parotid gland before suturing the overlying tissues provided our patient with an excellent functional and cosmetic result, without the formation of a fistula, sialocele, or Frey's Syndrome. CONCLUSION: We propose that the essential first step in the ideal closure of surgical wounds which penetrate the parotid gland involves the isolation and plication of the SMAS. This will create a barrier to the accumulation of parotid secretions which, in turn, may lead to the development of a sialocele and fistula formation. Such a barrier will also prevent the formation of Frey's Syndrome. Furthermore, plication of the SMAS serves to improve cosmesis by contributing soft tissue bulk to the wound and avoiding a concave deformity. PMID- 9865200 TI - Use of tissue meshing technique to facilitate side to side closure of large defects. AB - BACKGROUND: Primary closure of a large wound usually needs flaps of sophisticated design or skin grafts, both require more skill and wound care. Motley and Holt first reported the use of meshed advancement flap, a relative simple technique to close large defects of the lower leg in five patients. OBJECTIVE: To report the use of tissue meshing technique to close large wounds with significant tension on various sites of the body. METHODS: Six patients with large benign or malignant neoplasms at various sites of the body were included. Tissue meshing technique was used to facilitate wound closure after elliptical excision of the tumors at office visits. RESULTS: The wound defects, ranging from 3.0-3.5 cm in width, were closed with satisfactory cosmetic results, except for the occurrence of transient small hypertrophic scars in one patient. There was no complication of wound dehiscence, ischemia, infection or hematoma. CONCLUSION: Tissue meshing technique is a simple procedure and appears to be a satisfactory alternative to facilitate the closure of large wounds under tension. This technique is suitable for most body sites excluding central face and neck because there may be a stippled appearance in the area of meshing. PMID- 9865201 TI - Power boosting the grafts in hair transplantation surgery. Evaluation of a new storage medium. AB - BACKGROUND: When performing hair transplantation procedures, it is of the foremost importance to try to obtain the maximum survival rate possible of transplanted micrografts. OBJECTIVE: Aim of this study was to evaluate, in an in vitro model, the effects of preserving micrografts, for five hours, in an enriched storage medium in order to enhance the survival rate of hair micrografts. METHODS: A total of 200 human anagen hair follicles was obtained from ten male patients. Follicles were thus randomly assigned to one of the following group: Group A (control; n = 100 follicles), preserved for five hours in saline, and Group B (experimental; n = 100 follicles), preserved for five hours in a storage medium, containing adenosine triphosphate-magnesium chloride and deferoxamine mesylate. Isolated hair follicles from both Groups were then cultured for 10 days. RESULTS: A statistically significant difference was found between the survival rate of experimental (98%) and control follicles (87%). CONCLUSION: In our opinion, a "metabolic preconditioning" of micrografts by means of storing them for 5 hours in the described medium may be of some utility in augmenting the survival rate of hair grafts when performing hair transplantation surgery. PMID- 9865202 TI - Tie-over bolster dressings may not be necessary to secure small full thickness skin grafts. AB - BACKGROUND: The accepted method for securing full thickness skin grafts (FTSG) is with a tie-over bolster dressing, with or without basting sutures. We question the need for tie-over bolster dressings for small FTSGs. OBJECTIVE: We describe our method of FTSG reconstruction and the outcome in a consecutive series. METHODS: Thirty patients with surgical defects following tumour excision from the face24, scalp1, ear2, and finger3 ranging in diameter from 8 to 45 mm (mean 20 mm) were reconstructed with a FTSG. Interrupted monofilament nylon perimeter sutures only were used, with antibiotic ointment at the wound edge with either a light dressing or no dressing. RESULTS: A series of 30 FTSG secured without a tie over bolster dressing or basting sutures is described, all with good to excellent long term results. In two cases there was early superficial necrosis of the graft, but ultimately 100% graft take. CONCLUSION: We have found tie-over bolster dressings and basting sutures to be unnecessary in our series of small FTSG. This technique saves time and material and minimizes handling of the graft. PMID- 9865203 TI - Expression of c-fos and c-Ha-ras proto-oncogenes is induced in human chronic wounds. AB - BACKGROUND: It is known that chronic wounds such as leg ulcers have an increased risk of malignant transformation, mainly to squamous cell carcinoma (reviewed in Reference 1). OBJECTIVES: To better understand the molecular changes responsible for this increased risk of malignant transformation, we examined the expression of c-fos and c-Ha-ras, proto-oncogenes known to be involved in cellular proliferation and transformation, in chronic wounds. RESULTS: The level of both c fos and c-Ha-ras mRNAs, determined by in situ hybridization, were heavily induced in the basal layer of epidermis in chronic wounds when compared to normal skin and acute wounds, (originated from the edge of wounds created by Moh's surgery). Expression of both proto-oncogenes was not detected in the dermis of any groups. CONCLUSIONS: These results suggest that elevated expression of proto-oncogenes may be partly responsible for the increase susceptibility of chronic wounds to malignant transformation. PMID- 9865204 TI - Basal cell carcinoma of the genitalia. AB - BACKGROUND: Basal cell carcinomas (BCC) arising on the genitalia are exceedingly rare with an unclear pathogenesis. OBJECTIVE: To better understand risk factors, tumor characteristics, and the possible role of human papillomavirus (HPV) in the development of BCC of the genitalia. METHODS: 1543 records of Mohs micrographic surgery performed during a 6-year period were reviewed to identify cases of BCC arising on the genitalia. Tumor tissue was analyzed for HPV DNA by in situ hybridization. RESULTS: Four patients with BCC of the genitalia were treated with Mohs micrographic surgery. The malignancies were located on the scrotum, perineum, and perianal areas in the three male patients and on the vulva in the female patient. The mean age was 67 years. None of the patients had prior history of skin cancers. Histologic evaluation of the tumors revealed two nodular subtypes, one superficial subtype, and one with follicular differentiation. In situ hybridization failed to reveal DNA of HPV types 6, 11, 16, 18, 30, 31, 33, 35, 45, 51, and 52. CONCLUSION: In this small series, genital BCC occurred in an older age group with no identifiable predisposing risk factors and did not show evidence of HPV infection. PMID- 9865205 TI - Persistent keratoacanthoma: challenges in management. AB - BACKGROUND: Keratoacanthoma is a common cutaneous neoplasm, although the persistent form is less common and often more difficult to manage. Multiple treatment approaches have been attempted with variable efficacy. Establishing the diagnosis and selecting a treatment plan for persistent keratoacanthoma is often challenging. OBJECTIVE: Our purpose is to describe the difficulty one may encounter in the diagnosis and treatment of persistent keratoacanthoma. Hopefully, review of this clinical conundrum may facilitate the management of the reader's future patients. METHODS: We describe a case of persistent keratoacanthoma where the diagnosis was initially elusive and the management challenging. Our thought process during each stage of diagnosis and management is described in the form of "issues" with references to the appropriate literature. RESULTS: After several diagnostic and therapeutic interventions, successful treatment was achieved with administration of oral isotretinoin. Long-term remission continued as the dosage was tapered. CONCLUSION: Persistent keratoacanthoma may be challenging to diagnose and manage, presenting a clinical conundrum. Careful review of the clinicopathologic presentation and an understanding of the various treatment options may result in a successful outcome. PMID- 9865206 TI - A new treatment for syringoma. Combination of carbon dioxide laser and trichloroacetic acid. AB - BACKGROUND: Although syringoma represents a benign tumor of skin appendage, multiple and diffuse facial lesions can cause cosmetic problems for the affected individuals. Scarring, recurrence and postinflammatory hyperpigmentation can be serious troubles, especially for oriental people receiving various therapeutic modalities. OBJECTIVE: The purpose of this study was to evaluate the histopathologic and clinical efficacy of a new therapeutic approach for syringoma, consisting of combination of carbon dioxide (CO2) laser and 50% trichloroacetic acid (TCA). METHODS: Eighty skin biopsy specimens (45 patients) were evaluated to determine the depth of syringoma lesion. Among these, 28 specimens were obtained after a single test-pulse of CO2 laser (focused, superpulsed mode, power setting of 0.7-2 watts, beam diameter of 0.1 mm, pulse duration of 0.1 seconds), and 3 specimens after one pass of CO2 laser and 50% TCA. The depths of laser-induced vaporization and TCA-induced tissue necrosis were measured. Twenty patients were treated with the combination therapy of CO2 laser (two passes of laser) and 50% TCA. Clinical effectiveness and complications of combination therapy were evaluated by direct observation and photographs. RESULTS: Tumor depth: Analysis of eight specimens revealed tumor depth of 0.70 +/ 0.20 mm (mean +/- SD), ranging from 0.4-1.2 mm. There were no correlations between tumor depth and age of onset or duration of the tumor. Laser penetration depth: CO2 laser irradiation, 0.7-2 watts, induced vaporization of 0.29 +/- 0.12 mm in depth with carbonized rim of 0.1 +/- 0.03 mm width (n = 28 from 15 patients). With laser power of 1 watt, vaporization ranged from 0.17-0.45 mm in depth. Laser and TCA induced necrosis: TCA application resulted in extra tissue necrosis of 0.22-0.25 mm in depth beyond the level achieved by laser vaporization. CLINICAL FINDINGS: Among the 20 patients treated with the combination therapy, therapeutic effect for 11 patients was excellent, 6 patients showed good clinical response, and 3 patients were fair. There were no serious complications resulting from this procedure such as infection, scarring or textural change. CONCLUSION: The application of 50% TCA after CO2 laser irradiation was effective for removing deep-seated syringoma cells and for reducing the side effects, especially scarring. To the best of our knowledge, this is the first study introducing the combination of CO2 laser and TCA for removal of syringoma. PMID- 9865207 TI - Spontaneous healing of Mohs wounds of the cheek: a cosmetic assessment. AB - BACKGROUND: Facial defects after Mohs surgery may be closed at the time of surgery, shortly thereafter, or allowed to heal spontaneously. Selection of appropriate management options requires accurate prediction of the final cosmetic result. OBJECTIVE: To determine the relative merits of treatment options for facial defects resulting from Mohs surgery. METHODS: We prospectively and retrospectively analyzed outcomes for 132 wounds of the cheek resulting from Mohs surgery and allowed to heal spontaneously. The cheek was delineated into topographic areas and the wound location, size, and depth were recorded. Patients were observed intermittently, and a final evaluation of cosmesis was made after > or = 6 months. RESULTS: Most wounds in the nasolabial fold and preauricular areas healed with excellent results and half the wounds in the medial area of the cheek healed with good or excellent results. Central and mandibular areas of the cheek healed unpredictably, and results were unsatisfactory for most of these wounds. CONCLUSION: Final cosmetic results of Mohs surgery in the cheek area can be predicted on the basis of location, size, and depth of the wound, enabling physicians to knowledgeably select the most suitable treatment option. PMID- 9865208 TI - The use of low energy photon therapy (LEPT) in venous leg ulcers: a double-blind, placebo-controlled study. AB - BACKGROUND: Venous ulcers are estimated to be present in 0.2 to 0.4% of the population. Although new therapies have significant promise, nonhealing ulcers still represent a significant problem. OBJECTIVE: To evaluate the efficacy of low energy photon therapy (LEPT) in the treatment of venous leg ulcers. METHODS: A placebo-controlled, double-blind study using low energy photon therapy was performed in nine patients with 12 venous ulcers. Treatment was given three times a week for 10 weeks, using two monochromatic optical sources. One source provided a wavelength (lambda) of 660 nm (red) while the second source delivered a wavelength of 880 nm (infrared). Two optical probes were used, one consisted of an array of 22 monochromatic sources, operating at a wavelength of 660 nm and covering an area 6 x 10 cm2. The second probe had seven infrared sources, operating at a wavelength of 880 nm and covering an area of 4 cm2. The above configuration of optical probes was selected to cover the majority of the ulcer area being treated. The patients who were randomized to placebo treatment received sham therapy from an identical-appearing light source from the same delivery system. RESULTS: Nine patients with 12 venous ulcers were randomized to receive LEPT or placebo therapy. At the conclusion of the study, the percentage of the initial ulcer area remaining unhealed in the LEPT and placebo groups was 24.4% and 84.7%, respectively (P = 0.0008). The decrease in ulcer area (compared to baseline) observed in the LEPT and placebo groups was 193.0 mm2 and 14.7 mm2, respectively (P = 0.0002). One patient dropped out of the study, complaining of lack of treatment efficacy; he was found to be randomized to the placebo group. There were no adverse effects. CONCLUSION: In this placebo-controlled, double blind study LEPT was an effective modality for the treatment of venous leg ulcers. PMID- 9865209 TI - Combination therapy of resistant warts in a patient with AIDS. AB - BACKGROUND: An AIDS patient on anti-HIV therapy consisting of two reverse transcriptase inhibitors had a low CD4+ lymphocyte count, a high HIV viral RNA load, and facial warts that were resistant to ablative therapy. OBJECTIVE: To find a regimen to treat resistant warts in an AIDS patient with a depressed CD4 count and high HIV viral load. METHODS: A protease inhibitor was added to his regimen and he was treated with ablative therapy using a 585 nm flashlamp-pumped pulsed dye laser (FPPDL). RESULTS: Observed resolution of the warts one month later, with no recurrence at seven-month follow-up. CONCLUSIONS: Treatment of resistant warts in patients with AIDS may be enhanced by triple-drug therapy which includes a protease inhibitor, used in combination with ablative therapy, such as the FPPDL. PMID- 9865210 TI - Skin resurfacing of facial rhytides and scars with the 90-microsecond short pulse CO2 laser. Comparison to the 900-microsecond dwell time CO2 lasers and clinical experience. AB - BACKGROUND: Carbon dioxide lasers that produce either short pulses or scanned continuous beams have been used for skin resurfacing to improve wrinkles or scars. Using a high peak power, short pulse CO2 laser can produce clinically effective results with minimal thermal damage. OBJECTIVE: To evaluate the effectiveness of skin resurfacing using the 90-microsecond pulse duration CO2 laser compared to other laser systems. Erythema, healing time, complications, and histological measurement of the depth of ablation and thermal damage per pass with this system were also assessed. METHODS: Forty-one patients with facial rhytides and scars underwent resurfacing with a 90 microseconds pulse duration CO2 laser. Using patient survey, patients were evaluated for effectiveness of therapy, healing time, and complication rates. Comparisons of histologic and clinical findings were made with different short pulse CO2 lasers. RESULTS: Healing time, duration of erythema, and post-operative pain were less with the 90 microseconds pulse CO2 laser than with the 900-microsecond dwell time and 950 microsecond pulse duration lasers, while effectiveness was comparable. Complications were few with the 90-microsecond pulse laser, including three patients (9.1%) developing hyperpigmentation. One pass with the 90-microsecond pulse duration CO2 laser produced 100 microns of ablation with 17 microns of thermal damage. Ablation and damage were additive so that, by six passes, ablation depth was 350 microns and depth of thermal damage was 63 microns. This thermal damage is less than that reported with lasers having a longer pulse duration or dwell time with comparable depths of vaporization. CONCLUSION: Treatment with the 90-microsecond pulse duration laser results in a more rapid healing time and shorter duration erythema. The clinical improvements in wrinkles and sun damage were comparable. The 90-microsecond pulse duration laser provides an effective, predictable, and safe means of improving facial rhytides and scars. PMID- 9865211 TI - A practical review of laser-assisted hair removal using the Q-switched Nd:YAG, long-pulsed ruby, and long-pulsed alexandrite lasers. AB - BACKGROUND: The recent development of numerous laser-assisted hair removal techniques has produced confusion in the field of cutaneous laser surgery. OBJECTIVES: A systematic review of the hair removal laser systems currently available is attempted. Appropriate patient selection, treatment protocols, and discussion of each system's advantages, disadvantages, and complications is provided. CONCLUSIONS: Techniques involving the use of selective Q-switched Nd:YAG and long-pulsed alexandrite and ruby laser systems provide a significant delay in hair growth. With further technologic advances and understanding of proper treatment parameters and intervals, it is expected that permanent hair removal may eventually be possible. PMID- 9865212 TI - Cutaneous vascular proliferation following ultrapulse CO2 laser resurfacing. PMID- 9865213 TI - Topical diflorasone ointment for treatment of recalcitrant, excessive granulation tissue. PMID- 9865214 TI - Benign familial pemphigus (Hailey-Hailey disease). Treatment with the pulsed carbon dioxide laser. AB - BACKGROUND: Benign familial pemphigus (BFP) is a chronic blistering disease with significant morbidity. Surgical methods are often needed to control flares in difficult cases. OBJECTIVE: To describe the response of BFP to vaporization with a pulsed carbon dioxide (CO2) laser. METHODS: A 38-year-old woman with chest and axillary involvement unresponsive to conventional therapy was treated with the UltraPulse 5000 Laser (Coherent Medical Group, Palo Alto, CA). After active sites of BFP showed good response to treatment, we treated uninvolved skin of the left axilla to assess the efficacy of prophylactic therapy. RESULTS: Treatment of affected areas, except biopsy sites, resulted in clearing of active lesions after 1-2 weeks. We noted striking sparing of the treated areas from developing subsequent disease. The region that was later treated prophylactically has shown minor, asymptomatic recurrence of BFP in less than 5% of the area treated over an 18-month follow-up period. CONCLUSION: The pulsed carbon dioxide laser is a useful modality in treatment of BFP. In our patient, prophylactic treatment led to near complete eradication of disease in the treated area. A controlled, larger study is needed to confirm our results, and to determine optimal laser parameters. Long-term effects and duration of remission remain to be determined. PMID- 9865215 TI - Cosmetic surgery may be hazardous ... for the doctor as well as the patient. PMID- 9865216 TI - Pushing good research to go farther. PMID- 9865217 TI - The effect of hospital volume on the in-hospital complication rate in knee replacement patients. AB - OBJECTIVE: To examine the effect of hospital volume on in-hospital surgical outcomes for knee replacement using six years of Medicare claims data. DATA SOURCES/STUDY SETTING: The data include inpatient claims for a 100 percent sample of Medicare patients who underwent primary knee replacement during 1985-1990. We supplemented these data with information from HCFA's denominator files, the Area Resource File, and the American Hospital Association survey files. STUDY DESIGN: We estimated the probability that a patient has an in-hospital complication in the initial hospitalization for the first primary knee replacement, using a Logit model, for three definitions of complication. The models controlled for hospital volume, other hospital characteristics, patient demographics, and patient health status. We tested for the endogeneity of hospital volume. DATA COLLECTION/EXTRACTION METHODS: A panel of two orthopaedic surgeons and two internists reviewed diagnosis codes to determine whether a complication was likely, possible, or due to anemia. After removing the few observations with bad or missing data, the final population has 295,473 observations. PRINCIPAL FINDINGS: The probability of a likely in-hospital complication declines rapidly from 53 through 107 operations per year, then levels off. Statistical tests imply that hospital volume is exogenous in this patient-level data. Complication rates increased steadily through the study period. Although obesity appeared to lower the probability of a complication, a counterintuitive result, further investigation revealed this to be an artifact of the claims data limit of listing no more than five diagnoses. Controlling for this restriction reversed the effect of obesity. CONCLUSIONS: Rather than uncontrolled expansion of knee surgery to small hospitals, decentralization to regional centers where at least about 50, and preferably about 100, operations per year are assured appears to be the optimal policy to reduce in-hospital complications. PMID- 9865218 TI - Patterns of coordination and clinical outcomes: a study of surgical services. AB - OBJECTIVE: To test the hypothesis that surgical services combining relatively high levels of feedback and programming approaches to the coordination of surgical staff would have better quality of care than surgical services using low levels of both coordination approaches as well as those surgical service using low levels of either coordination approach. STUDY SETTING: A study sample of 44 academically affiliated surgical services that are part of the Department of Veterans Affairs. STUDY DESIGN: In a cross-sectional analysis, surgical services were assigned to one of three groups based on their scores on feedback and programming coordination measures: high on both measures; high on one measure, low on the other; and low on both. Univariate and multivariate analyses were used to assess differences among these groups with respect to three quality indicators: risk-adjusted mortality, risk-adjusted morbidity, and staff perceptions of quality. DATA COLLECTION/EXTRACTION METHODS: Risk-adjusted mortality and morbidity came from an outcomes reporting program within the Department of Veterans Affairs that entails the prospective collection of clinical data from patient charts. Data on coordination practices and perceived quality came from a survey of surgical staff at each of the 44 participating surgical services. PRINCIPAL FINDINGS: The group of surgical services using high feedback and high programming had the best perceived quality. This group also had the lowest morbidity, but the difference was statistically significant with respect to only one of the two other groups: the group with low feedback and low programming. No significant group differences were found for mortality. CONCLUSIONS: Study results provide partial support for the hypothesis that high levels of feedback and programming should be combined for optimal quality of care. Study results also suggest that staff coordination is more important for improving morbidity than mortality in surgical services. PMID- 9865219 TI - Multiple outcome assessment in a study of the cost-effectiveness of clozapine in the treatment of refractory schizophrenia. Department of Veterans Affairs Cooperative Study Group on Clozapine in Refractory Schizophrenia. AB - OBJECTIVE: To develop new methods for combining results from multiple outcome domains and to demonstrate their application in a study of the cost-effectiveness of clozapine in treating hospitalized patients with refractory schizophrenia. DATA SOURCES/STUDY SETTING: Interview assessments, and administrative utilization and cost data, concerning 423 patients with refractory schizophrenia who had been hospitalized for 30-364 days during the year before study entry, at 15 VA medical centers. STUDY DESIGN: A 12-month double-blind trial compared clozapine (n = 205) and haloperidol (n = 218) in the treatment of refractory schizophrenia. DATA COLLECTION/EXTRACTION METHODS: Data from standard assessment instruments, gathered at baseline and at 6 weeks, and at 3, 6, 9, and 12 months, were used to develop a Composite Health Index for Schizophrenia, a measure that addresses outcome in six domains, weighted by patient or provider preferences. Cumulative improvement was estimated by computing the area under the improvement curve. This measure was then combined with cost data, reflecting consumption of societal resources to estimate incremental cost-effectiveness ratios. PRINCIPAL FINDINGS: Clozapine was significantly more effective than haloperidol on measures of symptoms (p = .02) and side effects (p < .0001), with nonsignificant trends in the positive direction on community role functioning (p = .06), family relationships (p = .23), social relationships (p = .30), and daily activities (p = .20). Clozapine was also more effective than haloperidol on the one-year cumulative Composite Health Index for Schizophrenia (p < .0001 for all weighting schemes). After converting this measure to a 0-1 Worst Health-Good Health Scale analogous to Quality Adjusted Life Years, clozapine was found to yield a small improvement of .049 Worst Health-Good Health Units as compared to an improvement of only .027 Units for haloperidol (p < .0001). Average annual costs were $2,733 lower for clozapine (95% C.I. = -$9,220 to $3,754). Although clozapine was significantly more effective than haloperidol, the summary cost-effectiveness ratio had a wide 95 percent confidence interval ranging from -$431,585 to $177,352. CONCLUSIONS: Methods demonstrate an approach to using conventional disease-specific measures to evaluate the cumulative effectiveness of novel treatments for psychotic disorders and for expressing their economic effect as cost-effectiveness ratios. Among high hospital users with refractory schizophrenia, clozapine is more cost-effective than standard treatment, although the magnitude of its effect is small and there is considerable uncertainty about the cost estimates. PMID- 9865220 TI - Provider choice and use of mental health care: implications for gatekeeper models. AB - OBJECTIVE: To examine the ways in which the costs of nonresidential mental health care depend on (1) the type of provider who initiates the treatment episode and (2) the level of cost sharing imposed on the patient. STUDY SETTING: The 1987 National Medical Expenditure Survey, a national probability sample of the U.S. civilian, noninstitutionalized population. DATA COLLECTION: Data were collected during four personal interviews conducted during 1987 and 1988. Key variables include the type of provider contacted at the beginning of treatment (psychiatrist, other physician, nonmedical mental health care specialist) and the cost (total actual payments from all sources) for the treatment episode. METHODS OF ANALYSIS: An episodic model of demand for mental health care is estimated using a two-step procedure. Multinomial probit analysis is first used to determine the factors that influence the choice of initial provider type. Right censored Tobit analysis is used to determine the factors that affect the costs of care, including the type of provider who initiates the care episode. PRINCIPAL FINDINGS: Results indicate that out-of-pocket price does significantly (p < .05) affect the patient's initial choice of provider type but that, after controlling for the endogeneity of provider choice, price is no longer significant in explaining overall treatment costs. After controlling for selection effects, care episodes initiated by nonspecialist physicians are found to be as expensive as those initiated by psychiatrists and significantly more expensive than episodes initiated by nonphysicians. CONCLUSIONS: The results suggest that nonmedical mental health care specialists may be more effective than physicians in controlling costs when used as case managers in the care of persons with mental illnesses. PMID- 9865222 TI - Single women and the dynamics of Medicaid. AB - OBJECTIVE: To investigate transitions in and out of Medicaid for a cohort of single adult women of childbearing age in order to address questions that arise as policymakers try to encourage transitions from welfare to work. DATA SOURCES: Longitudinal data from Waves 2 through 8 of the 1990 panel of the Survey of Income and Program Participation, a nationally representative survey of American adults covering May 1990-1992. STUDY DESIGN: We estimate a series of discrete time logit models with duration dependence to obtain transition probabilities among Medicaid, privately insured, and uninsured spells. Explanatory variables in the models include prior insurance history, income limits on Medicaid by state, and important socioeconomic and demographic characteristics. We use these models to characterize insurance spells for a cohort of single women. PRINCIPAL FINDINGS: Most Medicaid spells are relatively short. Over half end in a year or less; only one spell out of seven lasts longer than five years. Two-thirds of Medicaid disenrollees become uninsured. Former welfare recipients are prone to frequent changes in insurance status. In states with more generous income limits for AFDC, women stay on Medicaid longer, but they do not move into the program at a faster rate. CONCLUSIONS: Imposing time limits on Medicaid eligibility would affect only a small proportion of Medicaid spells but would eliminate a significant proportion of the caseload at a point in time. In considering changes in Medicaid that would encourage transitions from welfare to work and would alter the dynamics of Medicaid, policymakers need to consider how transitions both in and out of private insurance and Medicaid would be affected. PMID- 9865223 TI - Who is the marginal patient? Understanding instrumental variables estimates of treatment effects. AB - OBJECTIVE: To clarify the issues of generalizability arising from the use of instrumental variable (IV) methods to estimate treatment effects in nonexperimental medical outcome studies. DATA SOURCE: We generate Monte Carlo data designed to resemble typical data sets where detailed health status information is unavailable and the treatment assignment process is unobserved. The model used to generate our data makes the realistic assumption that unobservable health status characteristics of patients influence the treatment assignment process and the effectiveness of treatment. STUDY DESIGN: We use Monte Carlo data to illustrate the circumstances where IV estimates generalize to an unobservable patient subpopulation and those where IV estimates generalize to the entire patient population represented by the sample used in the analysis. We also simulate the effect of two policy changes that affect practice patterns. Further, we show that IV estimates are useful for predicting the effect of these changes on treatment effectiveness when the subpopulation to which the IV estimate refers is the same or very similar to the population whose treatment status is affected by the policy change. CONCLUSIONS: Health services researchers cannot take for granted that IV estimates generalize to the same population represented by the sample used for analysis. Instead, researchers must rely on their knowledge of clinical practice and theory regarding the treatment assignment process in interpreting their results and in predicting the effect of changes in practice patterns. PMID- 9865221 TI - Cost-effectiveness of assertive community treatment versus standard case management for persons with co-occurring severe mental illness and substance use disorders. AB - OBJECTIVE: To determine the cost-effectiveness of Assertive Community Treatment (ACT) in comparison to Standard Case Management (SCM) for persons with severe mental illness and substance use disorders. DATA SOURCES AND STUDY SETTING: Original data on the effectiveness and social costs of ACT and SCM that were collected between 1989 and 1995. Seven community mental health centers in New Hampshire provided both types of treatment. STUDY DESIGN: Persons with schizophrenia, schizoaffective disorder, or bipolar disorder and a concurrent substance use disorder were randomly assigned to ACT or SCM and followed for three years. The primary variables assessed were substance use, psychiatric symptoms, functioning, quality of life, and social costs. DATA COLLECTION METHODS: Effectiveness data were obtained from interviews at six-month intervals with persons enrolled in treatment and with their service providers. Social cost and service utilization data came from client reports; interviews with informal caregivers; provider information systems and Medicaid claims; law enforcement agencies; courts; and community service providers. PRINCIPAL FINDINGS: Participants in both groups showed significant reductions in substance use over time. Focusing on quality of life and substance use outcomes, ACT and SCM were not significantly different in cost-effectiveness over the entire three-year study period. Longitudinal analyses showed that SCM tended to be more efficient during the first two years but that ACT was significantly more efficient than SCM during the final year of the study. CONCLUSIONS: In an adequately funded system, ACT is not more cost-effective than SCM. However, ACT efficiency appears to improve over time. PMID- 9865224 TI - Medical outcomes study short form 36: testing and cross-validating a second-order factorial structure for health system employees. AB - OBJECTIVE: To test the factorial validity of the SF-36. DATA SOURCE: Sample data collected in 1995 and 1996 using telephone interviews with health system employees as part of a study of health status. METHODS OF ANALYSIS: Confirmatory factor analysis and structural equation modeling techniques were used to evaluate the data. PRINCIPAL FINDINGS: The results of this study suggest that (1) Mental Health and Physical Health are not independent; (b) Mental Health cross-loads onto Physical Health; (c) general health loads onto Mental Health instead of Physical Health; (d) many of the error terms are correlated; (e) the physical function subscale is not reliable across the samples or the "age" or "education" subgroups; and (f) the mental health subscale path from Mental Health is not reliable across some subgroups. This hierarchical factor pattern was replicated across both samples. CONCLUSIONS: This study supports the second-order factorial structure of the SF-36. Adding the covariance path between the variables Physical Health and Mental Health improved model fit. Two paths from the second-order latent variables to the first-order latent variables differ from the original hypothesized structure of the SF-36. Health perception was influenced by Mental Health rather than Physical Health, and mental health was influenced by both Mental Health and Physical Health. This cross-loading suggests that the perception of Physical Health greatly affects mental health. Scale instabilities in the SF-36 across subgroups suggest that a comparison of mean scores or summary scores is inappropriate. Data interpretation can be improved if multigroups structural equation modeling is used. PMID- 9865225 TI - A new measure of contemporary life stress: development, validation, and reliability of the CRISYS. AB - OBJECTIVE: To develop and validate a measure of contemporary life stressors. STUDY SETTING: Three interview studies: Study 1 (pilot), 32 caregivers receiving case management services for a child with chronic illness; Study 2 (validation), 311 caregivers of children receiving general pediatric care at a university clinic; Study 3 (reliability), 17 caregivers of children with a complex medical diagnosis. STUDY DESIGN: Study 1: item development via discussions with case managers; piloted with caregivers. Study 2 examined psychometric properties of the measure and correlated it with the CES-D, a measure of depressive symptomatology and the PRQ85-Part 2, a measure of perceived social support, to establish its convergent construct validity. Study 3 established the test-retest reliability of the measure over two weeks by correlating two administrations of the index. DATA COLLECTION: Face-to-face interviews in homes (Study 1) or in clinic waiting rooms (Studies 2 and 3) and by telephone (Study 3 retest). PRINCIPAL FINDINGS: The CRISYS is a flexible, multidimensional tool that demonstrates strong face, content, and construct validity, and excellent test retest reliability. The format is easy to use and well accepted by respondents and is suitable for low-income populations. CONCLUSIONS: Researchers will find the CRISYS useful when evaluating the success of a clinical model or a healthcare system, and the effectiveness of an insurance plan or a government program. Clinicians may also find that the CRISYS is an effective screen for family needs. PMID- 9865226 TI - Increasing consolidation in healthcare markets: what are the antitrust policy implications? AB - OBJECTIVE: To discuss the implications of the rapid transformation of the healthcare financing and delivery system for competition, social welfare, and antitrust policy. PRINCIPAL FINDING: Horizontal and vertical consolidations can enhance efficiency but can also be anticompetitive in markets characterized by entry barriers. RECOMMENDATION: Active enforcement of the antitrust laws is essential to ensure that competition in healthcare markets will lead to procompetitive, rather than anticompetitive effects. However, healthcare antitrust enforcement policy must be flexible enough to allow efficient new forms of organization and practice to emerge. PMID- 9865227 TI - Defining competition in markets: why and how? AB - OBJECTIVE: To examine the variety of perspectives from which to study the measurement of competition in the healthcare marketplace. Based on a meeting held by The Robert Wood Johnson Foundation in 1996, the authors discuss the complications inherent in the way markets and products are defined by key stakeholders, including economists, policymakers, federal antitrust officials, purchasers, and the competitors themselves. CONCLUSION: The consensus among those who study this issue is that the way competitors, markets, and geographic areas are currently defined, and the ways of measuring competition, are inadequate, due mainly to the fact that both the measures and the definitions have been constructed from very limited data. Confounding this is the fact that analyses of competition are undertaken for such a wide variety of uses and that creating one database to solve the problems mentioned can be extremely daunting. RECOMMENDATIONS: Future research should examine ways to develop better definitions of the new healthcare structures that are competing with each other and ways to create measures of competition that include these new structures. To remedy gaps in the ability to measure competition, the field might also benefit from a public use data file, similar to the Area Resource File (ARF), that would contain HMO data according to geographic area, as well as provider data, employer data, payer data, and sociodemographic data. PMID- 9865228 TI - Managed care, market power, and monopsony. AB - OBJECTIVE: To examine the theoretical possibility of monopsony behavior under managed care insurance. STUDY DESIGN: Use of microeconomic theory to examine how managed care plans with market power would be expected to behave, and effects of that behavior on consumer and supplier welfare. PRINCIPAL FINDINGS: The article shows that, under managed care monopsony, the welfare of consumers may be increased but overall economic welfare will necessarily be reduced. It offers a test for whether the lower prices paid by managed care buyers with larger market share represent welfare-reducing monopsony or a welfare-increasing movement away from provider monopoly. The test says that, if the quantity of inputs (supplied under conditions of increasing long-run marginal cost) declines, monopsony is present. The article also argues that the translation of lower provider prices into lower premiums is consistent with welfare-reducing monopsony by nonprofit health plans. In contrast, for-profit health plans that obtain monopsony may reduce the welfare of consumers as well as that of input suppliers. These theoretical conclusions are shown to be consistent with recent empirical research indicating a negative relationship between buyer market power and cost per enrollee. CONCLUSIONS: Traditional antitrust policy has not been able to deal well with monopsony. The article concludes that health plans that use their market power to reduce medical spending may harm the well-being both of specialized medical workers and of consumers of medical care. Antitrust policy may need to be modified to deal with this situation. PMID- 9865229 TI - Marshfield Clinic, physician networks, and the exercise of monopoly power. AB - OBJECTIVE: Antitrust enforcement can improve the performance of large, vertically integrated physician-hospital organizations (PHOs). Objective: To examine the recent court decisions in the Blue Cross and Blue Shield United of Wisconsin v. Marshfield Clinic antitrust case to understand better the benefits and costs of vertical integration in healthcare. SUMMARY AND CONCLUSIONS: Vertical integration in the Marshfield Clinic may have had the benefits of reducing transactions and uncertainty costs while improving the coordination between ambulatory and inpatient visits, but at the cost of Marshfield Clinic's monopolizing of physician services and foreclosing of HMO entry in northwest Wisconsin. The denial of hospital staff privileges to non-Marshfield Clinic physicians combined with certificate-of-need regulations impeded physician entry and solidified Marshfield Clinic's monopoly position. Enforcement efforts of recent antitrust guidelines by the U.S. Department of Justice and the Federal Trade Commission will need to address carefully the benefits and costs of vertically integrated systems. PMID- 9865230 TI - Antitrust enforcement in the healthcare industry: the expanding scope of state activity. AB - OBJECTIVE: To describe the landscape of state antitrust activity and review related research and policy issues. In particular, to examine state laws that attempt to immunize mergers among healthcare providers from state and federal antitrust prosecution and consent decrees issued by state attorneys general permitting healthcare providers to merge. DATA SOURCES: State laws attempting to immunize collaborative activities from state and federal antitrust prosecution and consent decrees between state attorneys general and collaborating healthcare providers. PRINCIPAL FINDINGS: State antitrust agencies have been more willing than federal antitrust agencies to approve mergers that are contingent on the fulfillment of specific conditions that require continued oversight. CONCLUSIONS: Research is needed to inform policymakers about the consequences of state approved mergers on market performance. PMID- 9865232 TI - The spread of state any willing provider laws. AB - OBJECTIVE: To describe the growth of any willing provider (AWP) and freedom of choice (FOC) laws applicable to managed care firms and to explore empirically the determinants of their enactment. STUDY SETTING: A 1996 compendium of state laws and state-level data from the 1991-1994 period. STUDY DESIGN: Pooled cross section time-series logistic regression of the decision to enact various types of AWP and FOC laws. Analysis uses a public choice framework to examine enactment. Key variables include proxy measures of proponent and opponent strength and the political environment. PRINCIPAL FINDINGS: The model works well for laws affecting hospitals, but performs poorly for physician and pharmacy laws. More providers are associated with the enactment of AWP and FOC laws. More large employers are associated with a reduced likelihood of enactment of some forms of the laws but not others. Conservative states are more likely to enact laws limiting selective contracting with hospitals and physicians. States with greater interparty competition are also more likely to adopt some types of legislation. CONCLUSIONS: The empirical results generally are consistent with the view that AWP and FOC laws are often enacted as a defensive strategy on the part of providers, but additional research is needed to provide a more definitive assessment of the determinants of these laws. Suggestions for future research are provided. PMID- 9865233 TI - Glycolate kinetics and hemodialysis clearance in ethylene glycol poisoning. META Study Group. AB - OBJECTIVE: Toxic manifestations following ethylene glycol exposure are due to accumulation of metabolites, particularly glycolate. We characterized glycolate elimination kinetics and dialysis properties in a series of ethylene glycol poisonings. METHODS: Patients who ingested ethylene glycol and received fomepizole (4-methylpyrazole; 4-MP) +/- hemodialysis were prospectively evaluated. Serial blood samples for ethylene glycol, glycolate, pH, and bicarbonate were drawn to determine glycolate elimination rate, t1/2, and correlations between initial glycolate and initial markers of acidosis. Dialyzer inlet and outlet samples were obtained to measure hemodialysis glycolate clearance. Plasma ethylene glycol and glycolate were determined by gas chromatography. RESULTS: Ten patients, mean age 49 years (range 28-73 years), presented a mean of 10.5 hours (range 3.5-21.5 hours) after ethylene glycol ingestion. Mean initial ethylene glycol was 18.5 mmol/L (range 0.8-62.2 mmol/L) (115 mg/dL; range 5-386 mg/dL) and glycolate was 17.0 mmol/L (range 10.0-23.7 mmol/L). Nine of 10 underwent hemodialysis. Nonhemodialysis (n = 4) elimination rate was 1.08 +/- 0.67 mmol/L/h (mean +/- SD) and t1/2 was 626 +/- 474 minutes. Elimination t1/2 during hemodialysis (n = 8) was 155 +/- 42 minutes. Hemodialysis clearance (n = 5) was 170 +/- 23 mL/min with flow rates 250-400 mL/min. Pearson correlation coefficients were: anion gap vs glycolate r2 = 0.65 (p = 0.005), bicarbonate vs glycolate r2 = 0.10 (NS) and pH vs glycolate r2 = 0.06 (NS). CONCLUSION: Glycolate has a slow elimination rate and long half-life. Hemodialysis effectively clears glycolate. An increased anion gap correlates with the presence of glycolate. Hemodialysis is projected as useful for ethylene glycol-poisoned patients with anion gap acidosis and low ethylene glycol blood levels. PMID- 9865231 TI - Nonprofit conversion: theory, evidence, and state policy options. AB - OBJECTIVE: To describe the contributions of nonprofit hospitals and health plans to healthcare markets and to analyze state policy options with regard to the conversion of nonprofits to for-profit status. DATA SOURCES/STUDY SETTING: Secondary national and state data from a variety of sources, 1980-present. STUDY DESIGN: Policy analysis. DATA COLLECTION/EXTRACTION METHODS: Development of a conceptual economic framework; analysis of empirical, legal, and theoretical literature; and review of statutes, rules, and court decisions. PRINCIPAL FINDINGS: Three main rationales support special status for nonprofits, especially hospitals: charity care, other community benefits, and consumer protection. The main social rationale for for-profits is their incentives for better efficiency. There are reasons to expect that nonprofit and for-profit goals differ; however, measured differences in community hospital cost, prices, and quality between nonprofit and for-profit hospitals are undetectable or inconclusive. Nonprofit hospitals do provide more uncompensated care than for-profit hospitals. Similarities between nonprofit and for-profit hospitals may exist because nonprofits may set norms that for-profits follow to some degree. States have substantial power and discretion in overseeing nonprofit conversions. Some have regularized oversight through new legislation that constrains, but does not eliminate, state officials' discretion. These statutes may be deferential to converting entities and their buyers or may be very restrictive of them. CONCLUSIONS: Overseeing the appropriate disposition of nonprofit assets in individual conversions is extremely important. States should also monitor local market conditions through community benefits assessments and other data collection, however, to accurately assess (and possibly redress) what is lost or gained from conversion. Local market conditions are likely more important in determining hospital behavior than ownership form. Potentially, a mix of for profit and nonprofit hospitals in a given market may improve market performance due to constraints the two ownership types may exercise over one another. If nonprofits disappear, the states may need to maintain quality and access norms through regulation. PMID- 9865234 TI - The effects of nutrition on plasma cholinesterase activity and cocaine toxicity in mice. AB - BACKGROUND: Low plasma cholinesterase activity is associated with severe cocaine toxicity in human subjects and animal experiments. Exogenously enhanced plasma cholinesterase activity is protective against cocaine toxicity in animals. Cocaine users tend to have lower plasma cholinesterase activity than controls. Yet, when cocaine users are allowed to use cocaine in controlled settings without dietary restriction, their plasma cholinesterase activity increases. This study evaluates the influence of diet on plasma cholinesterase activity and cocaine toxicity. METHODS: Forty-five Swiss albino mice were maintained on a high (30%) protein diet for 3 weeks. They were then randomized into equal groups and given either the high protein diet, an isocaloric low protein diet, or a protein and calorie deficient diet which consisted of reduced intake of the high protein diet. Body weights and plasma cholinesterase activities were measured after a 21 day study period. All animals then received a fixed dose of intraperitoneal cocaine and were observed for seizures and death. RESULTS: Body weights and plasma cholinesterase activities of the high protein animals remained stable. Weights for the low protein and reduced intake animals fell by 5% and 15%, respectively (p < 0.05 for both vs baseline). Similarly, plasma cholinesterase activities for the low protein and reduced intake animals fell by 4% and 10%, respectively (p = 0.06 for low protein and < 0.05 for reduced intake vs baseline). Cocaine caused seizures in 67% of the high protein animals as compared to 93% and 100% of the low protein and reduced intake animals, respectively (p < 0.05 for high protein vs reduced intake). None of the high protein animals died as compared to 20% and 100% of the low protein and reduced intake animals, respectively (p < 0.05 for high protein vs reduced intake). CONCLUSION: Protein and calorie malnutrition is associated with a reduction in plasma cholinesterase activity and enhanced cocaine toxicity in mice. Further study is needed to determine if dietary factors are partially responsible for variations in plasma cholinesterase activity and cocaine susceptibility in humans. PMID- 9865235 TI - The clinical significance of hyperamylasemia in organophosphate poisoning. AB - OBJECTIVE: Hyperamylasemia with a presumptive diagnosis of acute pancreatitis has been reported following organophosphate poisoning but there are no large-scale studies incorporating more specific diagnostic criteria. METHODS: Retrospective review of the medical records of 159 patients with a diagnosis of organophosphate poisoning over 3 years. Serum amylase, pancreatic amylase, salivary amylase, lipase and cholinesterase levels, and the clinical manifestations were analyzed. RESULTS: Serum amylase data was available for 121 of the 159 study patients. Hyperamylasemia (amylase > or = 360 U/L) was found in 44 patients (36%). Lipase was measured in 28 patients with hyperamylasemia; 9 of 28 had hyperlipasemia (lipase > or = 380 U/L). The finding of hyperamylasemia was closely related to clinical severity and presence of shock. A presumptive diagnosis of painless acute pancreatitis was diagnosed by hyperlipasemia associated with hyperamylasemia, clinical severity, serum LDH, and leukocyte counts. Two patients with presumptive pancreatitis died. Shock, coma, and hypoalbuminemia were the factors predicting fatality. CONCLUSIONS: Hyperamylasemia is frequent in severe organophosphate poisoning. However, hyperamylasemia is not synonymous with acute pancreatitis and pancreatic amylase is not a reliable parameter in the diagnosis of organophosphate-induced pancreatitis due to its low sensitivity and specificity. Lipase assay is indicated in patients with hyperamylasemia for early diagnosis of pancreatitis. Proper image studies and even pathological examination are also needed to confirm the extent of pancreatic injury. With prompt diagnosis and appropriate treatment, a complete recovery can be anticipated unless the patient has otherwise unrelated complications. PMID- 9865236 TI - Randomized placebo-controlled trial of 2,3-dimercaptosuccinic acid in therapy of chronic arsenicosis due to drinking arsenic-contaminated subsoil water. AB - INTRODUCTION: Chronic arsenic toxicity producing various clinical manifestations is currently epidemic in West Bengal, India, Bangladesh, and other regions of the world. Animal studies have indicated that 2,3-dimercaptosuccinic acid can be used as an oral chelating agent. A prospective, double-blind, randomized controlled trial was carried out to evaluate the efficacy and safety of 2,3 dimercaptosuccinic acid for chronic arsenicosis due to drinking arsenic contaminated (> or = 50 micrograms/L) subsoil water in West Bengal. METHOD: Twenty-one consecutive patients with chronic arsenicosis were individually randomized (random number; assignment made by individual not evaluating patients) into 2 groups: 11 patients (10 male, age 25.5 +/- 8 years) received 2,3 dimercaptosuccinic acid 1400 mg/d (1000 mg/m2) in the first week and 1050 mg/d (750 mg/m2) during the next 2 weeks with a repeat course 3 weeks later. The other 10 patients (all male, age 32.2 +/- 9.7 years) were given placebo capsules for the same schedule. The clinical features were evaluated by an objective scoring system before and after treatment. Routine investigations including liver function tests, arsenic concentrations in urine, hair, and nails, and skin biopsy evaluations were also completed. RESULTS: Though there was improvement in the clinical score of 2,3-dimercaptosuccinic acid-treated patients, similar improvement was observed in the placebo-treated group. There were no statistical differences in the clinical scores between the 2 groups at the beginning and at the end of treatment. Similarly, no differences were found for the other investigated parameters. CONCLUSION: Under the conditions of this study, 2,3 dimercaptosuccinic acid was not effective in producing any clinical or biochemical benefit or any histopathological improvement of skin lesions in patients with chronic arsenicosis. PMID- 9865237 TI - Isotopic ratio analysis in residential lead-based paint and associated surficial dust. AB - OBJECTIVES: This investigation assessed the contribution of lead in lead-based paint (7 samples) to lead-laden dust (8 samples) in a single suburban vacant residence using isotopic ratio analysis. METHODS: Interior/exterior lead-based paint surface concentration was measured by X-ray fluorescence while dust and scrapings were analyzed chemically for total lead content and by mass spectrometry for the associated isotopic ratios. RESULTS: Four out of 5 comparisons of paint (7 samples) and dust (8 samples) for a given location did not match isotopically. In the one location where the isotopic ratio of the paint and dust samples matched closely, some portions of the paint were not intact. One explanation for the isotopic ratio match is that the dust sample may have actually been contaminated with paint flecks. This explanation appears likely since the isotopic ratio for the lead in the dust and paint sample were not in the modern average range of US environmental lead, strongly indicating a local point source of the lead in this dust sample, namely the paint at this location. Lead dust samples whose isotopic ratio lies in the modern average range for US environmental lead cannot be correlated to the paint which is beneath them, since the isotopic ratio of lead in the dust may actually be a composite of many sources of lead over time, as suggested by an isotopic ratio in the modern average range. CONCLUSIONS: From the samples from this one house, the data dispute the contention that intact lead-based paint chalks and creates lead contaminated dust on its surface. While leaded household dust may contribute to children's lead exposure, intact paint need not contribute to surficial lead laden dust. Isotopic ratio measurements can be useful for point-source determination by virtue of sample match and by placement of the ratio on the spectrum of isotopic ratio values for lead. Point-source assessment based on isotopic ratio was either strengthened or weakened by placement outside or within the average range for US environmental lead, respectively. PMID- 9865238 TI - Isotope ratios and the source of lead in lead poisoning. PMID- 9865239 TI - Prolonged elimination half-life of phenol after dermal exposure. AB - BACKGROUND: Phenol is a general protoplastic poison which has been in use in medicine and industry for decades. It is readily absorbed through the skin causing both local and systemic toxicity. CASE REPORT: A 47-year-old male had 90% phenol spilled over his left foot and shoe (3% of body surface area). After a 4 1/2-hour exposure, manifestations included confusion, vertigo, faintness, hypotension, ventricular premature beats, atrial fibrillation, dark-green urine, and tense swelling, blue-black discoloration, hypalgesia, and hypoesthesia of the affected area. Treatment consisted of irrigation with copious amounts of water, incisions, and supportive measures. RESULTS: Peak serum phenol was 21.6 micrograms/mL, considered in the fatal range. Peak urine phenol plus urine conjugated phenol was 13,416 mg/g creatinine, indicating a major absorption. Elimination half-life was 13.86 hours, considerably longer than previously reported. CONCLUSIONS: Prolonged skin contact with concentrated phenol in an occlusive environment may result in a major absorption and a long elimination half-life even if the area involved is small. Prolonged elimination may be explained by extensive tissue distribution or by "slow-release reservoir" properties of the skin. Such exposure may be associated with severe systemic and local toxicities. Immediate removal from exposure and aggressive decontamination of the skin are essential to reduce these risks. PMID- 9865240 TI - Benzonatate overdose associated with seizures and arrhythmias. AB - BACKGROUND: Benzonatate is an antitussive with a unique chemical structure. It can contain as many as 8 structural analogs. Therefore, laboratory analysis of benzonatate is difficult. We report 2 cases of benzonatate poisoning with seizures and cardiac arrest and an analytical method to identify and quantify benzonatate in human plasma. CASE REPORTS: Case 1: A 12-month-old male presented to the emergency department of a rural hospital following ingestion of an unknown amount of benzonatate. Upon arrival, the child was seizing and in full cardiac arrest. Resuscitative measures were unsuccessful and the child died shortly after arriving at the emergency department. Case 2: A 39-year-old male ingested 36 benzonatate capsules in a suicide attempt. Enroute to the health care facility, the patient experienced a seizure, had a cardiac arrest, and was cardioverted. Upon arrival at the emergency department, the patient was acidotic with a pH of 6.8. Gastric lavage was performed followed by the administration of activated charcoal. Six hours after arrival at the emergency department, the patient was alert, oriented, and hemodynamically stable. The patient was observed for 24 hours and subsequently discharged. Laboratory confirmation of benzonatate in the plasma of the patient was performed using high-pressure liquid chromatography with tandem mass spectrometry (MS/MS). The benzonatate concentration was estimated to be 2.5 micrograms/mL. CONCLUSION: Seizures and cardiac arrest are possible following an acute ingestion. Quantitative analysis of benzonatate is possible using high-pressure liquid chromatography with tandem mass spectrometry. Routine analysis for benzonatate is not common. PMID- 9865241 TI - The use of pamidronate for hypercalcemia secondary to acute vitamin D intoxication. AB - INTRODUCTION: Pamidronate disodium, a bone resorption inhibitor through osteoclast mediation, has been used to treat chronic hypercalcemia secondary to malignancy and chronic renal failure. We report the use of pamidronate for acute, severe hypercalcemia secondary to iatrogenic vitamin D poisoning. CASE REPORT: A 77-year-old female nursing home resident was inadvertently administered 50,000 units of oral vitamin D daily for 6 days. The patient presented with lethargy, abdominal pain, and vomiting. The patient's initial serum calcium concentration was 5.25 mmol/L (21 mg/dL). The patient was initially treated with hydration and furosemide but developed congestive heart failure. Pamidronate was used and calcium concentrations normalized by 24 hours after treatment. CONCLUSION: We report a case of the use of pamidronate for significant hypercalcemia secondary to acute vitamin D poisoning. Although evidence of congestive heart failure was evident, dialysis was avoided without significant sequelae. Pamidronate therapy should be considered in patients with hypercalcemia secondary to acute vitamin D poisoning. PMID- 9865242 TI - Hemodialysis following butoxyethanol ingestion. AB - CASE REPORT: This case report describes a 19-year-old male who ingested a product containing butoxyethanol, propylene glycol, and monoethanolamine. Neurotoxicity, acidosis, and butoxyacetic acid persisted after hemodialysis. Hemodialysis was used to treat the acidosis, but the half-life of butoxyacetic acid did not appear to have been significantly altered. Fomepizole, a recently approved alcohol dehydrogenase inhibitor for ethylene glycol intoxication, was not available at the time of this case. Residual neurological deficits persisted after recovery from this severe intoxication by a glycol ether. PMID- 9865243 TI - Survival after massive ecstasy overdose. AB - INTRODUCTION: The toxicity profile of the amphetamine derivative 3,4 methylenedioxymethamphetamine (MDMA, "Ecstasy") is well known. This designer drug is usually taken at "house parties" and may cause severe complications, sometimes leading to death, even when taken in relatively small units (1 or 2 tablets). Up to now, only a few cases of survival after ingestion of an overdose of Ecstasy have been described. In most cases the users developed hyperthermia, disseminated intravascular coagulation, rhabdomyolysis, and renal failure. CASE REPORT: We describe a man who, after ingesting 50 tablets of Ecstasy (in combination with oxazepam and alcohol) at home, recovered within 2 days. Presenting features were unconsciousness, apnea, and convulsions. It is suggested that in most cases severe 3,4-methylenedioxymethamphetamine toxicity results from an interaction between direct pharmacological effects of the drug and the prevailing environmental conditions (high ambient temperature, dancing in trance, little fluid intake). PMID- 9865244 TI - Accidental, intravenous infusion of a peanut oil-based medication. AB - OBJECTIVES: To describe a case of fat embolus syndrome with lipoid pneumonia resulting from intravenous infusion of lipid and to illustrate the potential for accidental intravenous administration of vegetable oil-based progesterone preparations in the treatment of oncology patients. CASE REPORT: A patient with recurrent ovarian carcinoma accidentally received approximately 20 mL (0.29 mL/kg) of a peanut oil-based methylprogesterone product intravenously via infusion pump over 24 hours. The patient developed a lipoid pneumonia with dyspnea, cough, hypoxia, radiographic infiltrates, and a pleural effusion. She was hospitalized for 4 days, and signs and symptoms resolved over 2 weeks following steroids and supportive care. DISCUSSION: Experience with accidental or intentional intravenous lipid overdose in humans is limited. Typical findings of fat embolus syndrome are similar to lipid aspiration, with respiratory distress, hypoxia, and pulmonary infiltrates. In contrast to aspiration, however, fat embolus syndrome results in lipogranulomas surrounding blood vessels, rather than air passages, and potentially produces cerebrovascular, accident-like symptoms. Management of fat embolus syndrome is similar to that for lipid aspiration. However, as seen in this case, fat embolus syndrome typically resolves over several weeks as opposed to the 3-month to 1-year period seen with aspiration lipoid pneumonias. CONCLUSIONS: Accidental intravenous infusion of vegetable oil based products is a potential complication of the increased use of intravenous progesterones. PMID- 9865245 TI - Toxicological deaths of major league baseball players. AB - REPORT: A retrospective review of major league baseball records was conducted for players' cause of death. Any death attributed to a toxic exposure was analyzed for causal agent, reason for exposure, age at time of death, location, time of year, team, and dominant position played while active. RESULTS: Twenty-eight poisoning deaths were identified between 1889-1995. The most common agent was carbon monoxide (8), followed by methane gas asphyxiation (4), opiate overdose (4), ethanol (3), and phenol (3). Fourteen (50%) were unintentional deaths, 13 (46%) were suicidal in nature, and 1 (4%) homicidal. The majority of deaths (75%) occurred after the players had retired from the game. The leading position was pitcher (13), followed by catcher (5), outfield (4), second base (2), first (1), shortstop (1), third (1), and umpire (1). CONCLUSIONS: With society's increased illicit drug use, better drug detection, escalating salaries, and increased public pressures placed on present-day players, more poisonings may likely occur. PMID- 9865246 TI - Continuous flumazenil infusion in the treatment of zolpidem (Ambien) and ethanol coingestion. PMID- 9865247 TI - Indinavir overdose. PMID- 9865248 TI - Yew toxicity less than ipecac. PMID- 9865249 TI - Plantlore folklore. PMID- 9865250 TI - Is sulfasalazine toxic? PMID- 9865252 TI - The vaccine adverse event reporting system. PMID- 9865253 TI - Development of severe adverse effects after discontinuing amiodarone therapy in human heart transplant recipients. PMID- 9865254 TI - Effectiveness of immunosuppressive treatment for recurrent or refractory pancreas allograft rejection: correlation with histologic grade. PMID- 9865255 TI - Hepatitis B and C virus-induced diseases and acute rejection after liver transplantation. PMID- 9865256 TI - Cyclosporin impairs bile duct arterial supply in ischemic livers. PMID- 9865257 TI - Presence of IgG exhibiting a morphine-like activity in therapeutic intravenous immunoglobulins. PMID- 9865258 TI - Hypertriglyceridemia in renal transplant recipients treated with sirolimus. PMID- 9865259 TI - Monthly cyclophosphamide pulses in the treatment of crescentic glomerulonephritis. PMID- 9865260 TI - Conversion of stable renal transplant recipients from Sandimmune to Sang-35, a Neoral-equivalent cyclosporine formulation, using a dose-adjusted method. PMID- 9865261 TI - Concomitant detection by flow cytometry of the intragranular antigen granzyme B and the intranuclear antigen Ki-67 in peripheral blood mononuclear cells from healthy individuals and patients with acute CMV infection after renal transplantation. PMID- 9865262 TI - Irradiation or temporal variations in immunosuppressive schedules do not prevent graft-versus-host disease following rat limb allotransplantation. PMID- 9865263 TI - Outcome of liver transplantation after acute rejection and sepsis. PMID- 9865264 TI - Humoral and cell-mediated factors involved in the suppressive regulation induced by special blood derivatives and their clinical relevance. PMID- 9865265 TI - Increased systemic levels of soluble granzymes A and B during primary cytomegalovirus infection after renal transplantation. PMID- 9865266 TI - Selective expansion of a peripheral blood CD8+ memory T cell subset expressing both granzyme B and L-selectin during primary viral infection in renal allograft recipients. PMID- 9865267 TI - Docosahexaenoic and eicosapentaenoic acids inhibit the CD28-lymphocyte activation pathway in vitro. PMID- 9865268 TI - Malononitrilamides prevent and suppress allospecific antibody production. PMID- 9865269 TI - Possible role of post renal transplant graft-versus-host antibodies in preventing acute graft rejection. PMID- 9865270 TI - Allospecific T-cell receptor V-beta repertoire in human mixed lymphocyte cultures is conserved and oligoclonally restricted. PMID- 9865271 TI - Chronic allograft vasculopathy: new strategies for drug development. AB - Chronic rejection reflects cumulative trauma to the allograft regardless of its origin. The main histologic feature in chronic rejection in all allografts is proliferative allograft vasculopathy. According to the current paradigm, chronic rejection can be prevented by more intensive and selective immunosuppression. Experimental retransplantation to donor strain suggests that elimination of histoincompatibility does not prevent progression of the disorder after the initial stimulus has been sufficiently strong or long-lasting. In order to design additional sites for intervention after this point of no return, attention has focused on the regulation of the synthesis and/or action of smooth muscle cell growth factors. In several preclinical models, the blocking of growth factor synthesis, blocking of the binding of the growth factors to their receptors, and/or growth factor signaling downstream of the receptor, have successfully been used in the inhibition of the intimal response and allograft vasculopathy. Inhibition of proteolytic enzymes, necessary for smooth muscle cell locomotion, may provide additional sites of intervention. However, on most occasions, the inhibitory effect is incomplete only 30% to 50% of maximal. The future will demonstrate whether several approaches must be applied concomitantly, or whether genes regulating several of these ligands and their receptors simultaneously can be identified and exploited. PMID- 9865272 TI - Cytokine genotypes in allograft rejection: guidelines for immunosuppression. PMID- 9865273 TI - Chronic rejection: an update on the mechanism. PMID- 9865274 TI - Cytotoxic T lymphocytes and cytokines in graft vascular disease after clinical heart transplantation. PMID- 9865275 TI - Indirect allorecognition of donor HLA-DR peptides in chronic rejection of heart allografts. PMID- 9865276 TI - Adverse effects of immunosuppressive drugs on long-term graft outcome. PMID- 9865277 TI - Tolerance induction by megadose stem cell transplants: synergism between SCA-1+ Lin- cells and nonalloreactive T cells. PMID- 9865278 TI - Specific immunosuppression of the indirect recognition pathway. PMID- 9865279 TI - Short and long-term results after pretransplant high-dose single ATG-fresenius bolus in cadaveric kidney transplantation. PMID- 9865280 TI - In vitro functional properties of antithymocyte globulins: clues for new therapeutic applications? PMID- 9865281 TI - Dissociation between chimerism and skin graft tolerance after neonatal injection of allogenic spleen cells. PMID- 9865282 TI - Hematopoietic chimerism and tolerance of T cells, B cells, and NK cells. PMID- 9865283 TI - Deletion of donor-reactive cells followed by stem cell transplantation in recipients treated with total lymphoid-irradiation as a means for induction of transplantation tolerance to organ allografts and xenografts. PMID- 9865284 TI - A new large-animal model for research of graft vascular disease. PMID- 9865285 TI - On the horizon: clinical trials of new immunosuppressive strategies for autoimmune diseases. PMID- 9865286 TI - Evaluation of immunosuppressive induction regimens in renal transplantation. Collaborative Transplant Study. PMID- 9865287 TI - Immunoglobulin-cytokine fusion molecules: the new generation of immunomodulating agents. AB - Advances in the understanding of genetic engineering, protein expression, and Ig function have come together to allow for the rapid synthesis and production of a novel generation of potent immunomodulating reagents. Selective approaches that allow the isolation of desired specificities will be elucidated through meticulous engineering techniques. This, in turn, may eventually result in the fulfillment of the tremendous potential of engineered proteins for therapeutic and diagnostic applications. PMID- 9865288 TI - Effects of cyclosporine A monotherapy on the incidence of rejection and infection episodes in heart transplant patients. PMID- 9865289 TI - Intestinal metabolism of cyclosporine A in human heart transplant recipients. PMID- 9865290 TI - Evaluation of the AxSYM CyA assay compared with HPLC, TDx monoclonal and EMIT with both pretreatments. PMID- 9865291 TI - A meta-analysis to assess the safety and tolerability of two formulations of cyclosporine: Sandimmune and Neoral. PMID- 9865292 TI - A novel effect of cyclosporine A on lymphocyte migration to allograft and recipient lymphoid tissue. PMID- 9865293 TI - Plasma and low-density lipoprotein lipid peroxidation in cyclosporine A-treated children after liver transplant. PMID- 9865294 TI - Specific immunosuppression of the indirect recognition pathway. PMID- 9865295 TI - Enhanced efficacy of repeated anti-CD8 monoclonal antibody therapy by high-dose cyclosporine treatment. PMID- 9865296 TI - New trails in transplantation: how to exploit the potential of sirolimus in clinical transplantation. PMID- 9865298 TI - Therapeutic drug monitoring of tacrolimus in kidney transplantation: 9-month follow-up. PMID- 9865297 TI - Comparative efficacy of liposomal FK506 with FK506. PMID- 9865299 TI - Tacrolimus use in kidney-pancreas recipients is associated with less acute renal dysfunction than cyclosporine. PMID- 9865301 TI - Mycophenolate mofetil in ongoing chronic kidney allograft dysfunction. PMID- 9865300 TI - Tacrolimus (FK506) monotherapy provides potent and significant suppression of allogeneic foetal rat pancreatic allograft rejection. PMID- 9865302 TI - Mycophenolate mofetil immunosuppression in high-risk renal transplant recipients. PMID- 9865303 TI - A comparison of the safety and efficacy of mycophenolate mofetil, prednisone and cyclosporine and mycophenolate mofetil, and prednisone and tacrolimus. PMID- 9865304 TI - Pharmacoeconomic analysis of mycophenolate mofetil versus azathioprine in primary cadaveric renal transplantation. PMID- 9865305 TI - Comparison between mycophenolate mofetil and azathioprine based immunosuppression in pediatric renal transplantation from living related donors. PMID- 9865306 TI - Efficacy and safety of mycophenolate mofetil for the treatment of acute and chronic GVHD in bone marrow transplant recipient. PMID- 9865307 TI - Pharmacokinetics of mycophenolic acid in the early period after kidney transplantation. PMID- 9865309 TI - How to handle mycophenolate mofetil in combination with tacrolimus? PMID- 9865310 TI - Postmarketing evaluation of mycophenolate mofetil-based triple therapy immunosuppression compared with a conventional azathioprine-based regimen reveals enhanced efficacy and early pharmacoeconomic benefit after renal transplantation. PMID- 9865308 TI - Immunosuppression with cyclosporin A in combination with mycophenolate mofetil suppresses rejection of allogeneic fetal rat pancreatic allografts. PMID- 9865311 TI - A prospective randomised study of CSA monotherapy versus CSA plus mycophenolate mofetil in cadaveric renal transplant recipients. PMID- 9865312 TI - Long-term allograft survival and tolerance induction by the synergistic activity of malononitrilamides and tacrolimus. PMID- 9865313 TI - Preventive efficacy of 15-deoxyspergualin against progression of apoptotic cell death in acute rejection after rat liver transplantation. PMID- 9865314 TI - Functional characterization of HP12: a novel immunosuppressant purified from Helleborus species. PMID- 9865315 TI - In vitro transcription and translation of the tumour suppressor protein P53: qualitative and quantitative effects of FK506 and rapamycin. PMID- 9865316 TI - Effective oral ganciclovir prophylaxis against cytomegalovirus disease in heart transplant recipients. AB - The presented data show the combined sequential use of i.v. G for 14 days followed by PO G for 90 days is a much more effective prophylaxis for CMVD after heart transplantation than use of i.v. G for 14 days followed by PO A for 90 days. A need for hospitalization due to CMVD is significantly reduced by this new strategy. The follow-up in group II is shorter than in group I but is now at least 6 months in group II, without any new cases in the first 6 months after cardiac transplantation. Some currently unknown adverse effect of prolonged PO G, which may be present, is not identified in this analysis. PMID- 9865317 TI - Selective immunotherapy of IDDM: a discussion based on new findings from the RIP LCMV model for autoimmune diabetes. PMID- 9865318 TI - Protection of murine systemic lupus erythematosus by an I-E alpha-chain transgene. PMID- 9865319 TI - Strategies for the treatment of intraocular inflammatory disease. PMID- 9865320 TI - Anti-TNF alpha therapy is useful in rheumatoid arthritis and Crohn's disease: analysis of the mechanism of action predicts utility in other diseases. PMID- 9865321 TI - Bromocriptine markedly inhibits progression of murine retrovirus-induced immunodeficiency. PMID- 9865322 TI - FK-506 (tacrolimus) therapy for an unusual SLE-like disease. PMID- 9865323 TI - Prevention of experimental autoimmune diabetes in mice by treatment with leflunomide. PMID- 9865324 TI - Cyclosporine in the treatment of autoimmune disorders: a 10-year experience. PMID- 9865326 TI - Immunomodulatory activity of malononitrilamides, derivatives of leflunomide's primary metabolite, on models of experimental rheumatoid arthritis. PMID- 9865325 TI - Comparative study of the clinical efficacy of four DMARDs (leflunomide, methotrexate, cyclosporine, and levamisole) in patients with rheumatoid arthritis. PMID- 9865327 TI - Prevention of islet graft destruction in diabetic mice and rats by temporary anti IL-2 receptor therapy: comparison of different strategies. PMID- 9865328 TI - Leflunomide effectively treats naturally occurring immune-mediated and inflammatory diseases of dogs that are unresponsive to conventional therapy. PMID- 9865329 TI - Effects of leflunomide on experimental autoimmune myocarditis with special reference to the dendritic cells. PMID- 9865330 TI - Successful treatment of autoimmune hemolytic anemia with intravenous immunoglobulin in a patient with AIDS. PMID- 9865331 TI - Modulation of immunoglobulin dysregulation in graft versus host-and systemic lupus erythematosus-like diseases by malononitrilamides. PMID- 9865332 TI - Cyclosporine in the treatment of steroid-dependent and steroid-resistant idiopathic nephrotic syndrome in children. PMID- 9865333 TI - Monitoring serum beta-2 microglobulin is useful for detecting patients with increased risk of acute rejection during reduction in immunosuppression. PMID- 9865334 TI - Prolongation of concordant xenograft survival by treatment with FTY720. PMID- 9865335 TI - Xenogeneic rejection mechanisms shown by intravital microscopy. AB - The importance of this model is that it showed exactly where in the organ the xenogeneic damage occurred. The liver received the blood mainly via portal veins, which merge with the pulsatile arterioles in the Disse spaces. This periportal area is followed by the sinusoids and ends in the central or postsinusoidal vein. IVM enables us to differentiate between perfused and unperfused sinusoids and to calculate the ratio. Not all sinusoids are perfused at any time. It appears that 5% to 10% are unperfused. During xenoperfusion, only 65% of sinusoids show blood flow after a perfusion of 12 minutes. This is less than in hemorrhagic shock. Only the combined platelet inhibitors and apheresis resulted in remarkable improvement. The calculation of an index indicates the improvement of acinar perfusion. Thrombocytes and leukocytes remain, however, in the liver. In conclusion, the model used to analyze the dynamics of microvascular liver perfusion and sinusoidal perfusion is suitable for such investigations in a xenogeneic model. It has no major side effects, either on the perfusing blood or on the liver, as proved in the isogeneic control group. The important finding in our eyes is that the perfusion failure begins in the periportal fields, where the blood enters the foreign microvasculature and where the leukocytes first come in contact with the foreign endothelium. All previous manipulations had only a minor impact on this contact of cells with the foreign endothelium. The study indicates that the early events of xenogeneic hyperacute rejection are of unspecific character and involve leukocytes and thrombocytes to a major degree, thus being responsible for the dramatic decrease in the microcirculation in xenogeneic livers. PMID- 9865336 TI - Allogeneic and xenogeneic vascular injury and protection. PMID- 9865337 TI - Combination therapy of malononitrilamides and tacrolimus (FK 506) induced long term xenograft survival. PMID- 9865338 TI - Variable in vitro inhibition of HLA-specific alloantibody-mediated cytotoxicity by intravenous human immunoglobulin. PMID- 9865339 TI - Prolongation of rat fetal pancreas allograft survival using a nondepleting anti CD4 monoclonal antibody W3/25. PMID- 9865340 TI - Heme oxygenase inhibits nitric oxide synthase by degrading heme: a negative feedback regulation mechanism for nitric oxide production. PMID- 9865341 TI - Modulation of cytokine production following intranasal administration of antigenic peptide. PMID- 9865343 TI - Phytoalexin resveratrol (3-4'-5-trihydroxystilbene) modulates granulocyte and monocyte endothelial adhesion. PMID- 9865342 TI - Diltiazem modulates monokine production by isolated human monocytes. PMID- 9865344 TI - Positive effects of tacrolimus in a case of CIDP. PMID- 9865345 TI - Influence of clinical immunosuppression on proinflammatory cytokines and acute phase proteins. PMID- 9865346 TI - Immunosuppressant effect on Fas antigen expression and p53 of an experimental tumor. PMID- 9865347 TI - Inhibition of PDGF-stimulated rat smooth muscle cell proliferation by MNA 279 and MNA 715. PMID- 9865348 TI - Effect of WF10 (TCDO) on antigen presentation. PMID- 9865349 TI - Long-term cardiac allograft survival in mice treated with malononitrilamide 715. PMID- 9865350 TI - The nature of ischemia/reperfusion injury. PMID- 9865351 TI - Free radical generation in human endothelial cells exposed to anoxia and reoxygenation. PMID- 9865353 TI - Postischemic reperfusion injury and cytokines. PMID- 9865352 TI - Comment to the previous paper of J. Zweier: demonstration of reactive oxygen species in reperfused human kidney transplants. PMID- 9865355 TI - Cytokines 1: upregulation of cytokine gene expression and cytokine release in postischemic reperfused organs and transplants. PMID- 9865354 TI - Regulation of endothelial cell function by pro- and anti-inflammatory cytokines. PMID- 9865356 TI - Postischemic reperfusion injury and adhesion molecules in nontransplant models. PMID- 9865357 TI - Endothelial cell adhesion molecules in inflammation and postischemic reperfusion injury. PMID- 9865358 TI - Analysis of intragraft adhesion molecules and their release in clinical liver transplantation: impact of reperfusion injury. PMID- 9865359 TI - Upregulation of major histocompatibility complex-expression under ischemic conditions in experimental models. PMID- 9865360 TI - Possible role of postischemic reperfusion injury as initiator of allorecognition/alloactivation. PMID- 9865361 TI - Integrins and T helper cell activation. PMID- 9865362 TI - Postischemic reperfusion injury and allograft arteriosclerosis. PMID- 9865363 TI - Ischemia reperfusion injury and chronic allograft rejection. PMID- 9865364 TI - Decreasing the cold ischemia time. PMID- 9865365 TI - What is a reasonably short cold ischemia time in kidney transplantation? PMID- 9865366 TI - Cadaver kidney graft outcome in relation to ischemia time and HLA match. Collaborative Transplant Study. PMID- 9865367 TI - Organ procurement policy: should we reduce cold ischemia times? PMID- 9865368 TI - Notes on the Bellagio Task Force Report on Transplantation, Bodily Integrity, and the International Traffic of Organs. PMID- 9865369 TI - Expected costs for small-scale organ transplantation programs. PMID- 9865370 TI - Early catheter removal decreases incidence of urinary tract infections in renal transplant recipients. PMID- 9865371 TI - A trial of oral magnesium supplementation in renal transplant recipients receiving cyclosporine. PMID- 9865372 TI - Renal transplantation: potential source of microfilarial transmission. PMID- 9865373 TI - Increased dosage requirement and rejection after Neoral conversion in pediatric liver transplant patients. PMID- 9865374 TI - Liver disease in tropical renal transplant patients: an autopsy study. PMID- 9865375 TI - Massive ingestion of tacrolimus in a young liver transplant patient. PMID- 9865376 TI - Quality of life after liver transplantation. PMID- 9865377 TI - Analysis of pig-MAP after small bowel transplantation in pigs. PMID- 9865378 TI - Effect of low-molecular-weight heparin on development of cardiac allograft vascular disease following heart transplantation in rats. PMID- 9865379 TI - A reliable method for long-term (24-hour) hypothermic transport of cardiac graft. PMID- 9865380 TI - Mycophenolate mofetil and FK506: two novel immunosuppressants in murine corneal transplantation. PMID- 9865381 TI - Impaired hepatic mitochondrial function reserve in brain-dead pigs. PMID- 9865382 TI - Arterial ketone body ratio in clinical liver transplantation. PMID- 9865383 TI - Treatment of chronic viral hepatitis. AB - Interferon-alpha therapy has proved effective for up to 40% of patients with adult-acquired chronic hepatitis B virus (HBV) infection and for 20-25% of those with chronic hepatitis C virus (HCV) infection. Nucleoside analogues, such as lamivudine and famciclovir, are showing promise as antiviral agents for chronic HBV and the combination of interferon-alpha and ribavirin is proving to be successful therapy for 40-50% of patients with chronic HCV. In this article we review current therapy and discuss future strategies of the therapy of chronic viral hepatitis. PMID- 9865384 TI - Identification of HIV-1 integrase inhibitors based on a four-point pharmacophore. AB - The rapid emergence of human immunodeficiency virus (HIV) strains resistant to available drugs implies that effective treatment modalities will require the use of a combination of drugs targeting different sites of the HIV life cycle. Because the virus cannot replicate without integration into a host chromosome, HIV-1 integrase (IN) is an attractive therapeutic target. Thus, an effective IN inhibitor should provide additional benefit in combination chemotherapy. A four point pharmacophore has been identified based on the structures of quinalizarin and purpurin, which were found to be potent IN inhibitors using both a preintegration complex assay and a purified enzyme assay in vitro. Searching with this four-point pharmacophore in the 'open' part of the National Cancer Institute three-dimensional structure database produced 234 compounds containing the pharmacophore. Sixty of these compounds were tested for their inhibitory activity against IN using the purified enzyme; 19 were found to be active against IN with IC50 values of less than 100 microM, among which 10 had IC50 values of less than 10 microM. These inhibitors can further serve as leads, and studies are in progress to design novel inhibitors based on the results presented in this study. PMID- 9865385 TI - Synthesis, anti-human immunodeficiency virus activity and esterase lability of some novel carboxylic ester-modified phosphoramidate derivatives of stavudine (d4T). AB - We report the design, synthesis and antiviral evaluation of a series of lipophilic, masked phosphoramidate derivatives of the anti-human immunodeficiency virus (HIV) nucleoside analogue d4T, designed to act as membrane-soluble prodrug forms for the free nucleotide. In particular, we report a series of 12 novel compounds with systematic variation in the structure of the carboxylate ester function. In order to rationalize the changes in antiviral action with variation of this moiety we applied our recently developed 31P NMR-based assay for carboxyesterase lability to this series. However, no clear positive correlation emerged, indicating that, at least within this series, factors other than simple esterase lability may be the major determinants of antiviral potency. PMID- 9865386 TI - Regiospecific synthesis and anti-human immunodeficiency virus activity of novel 5 substituted N-alkylcarbamoyl and N,N-dialkylcarbamoyl 1,2,3-triazole-TSAO analogues. AB - Several 5-N-alkyl and 5-N,N-dialkylcarbamoyl substituted analogues of the anti human immunodeficiency virus (HIV) type 1 lead compound [1-[2',5'-bis-O-(tert butyldimethylsilyl)-beta-D-ribofuranosyl]-5- (N,N-dimethylcarbamoyl)-1,2,3 triazole]-3'-spiro-5"-(4"-amino-1",2"- oxathiole-2",2"-dioxide) have been prepared and evaluated as inhibitors of HIV-1 replication. A new regiospecific synthetic procedure is described. The compounds were prepared by cycloaddition of the appropriate glycosylazide to 2-oxoalkylidentriphenyl-phosphoranes, followed by treatment with primary or secondary amines, to yield, exclusively, 5 substituted 1,2,3-triazole-TSAO analogues. Several 5-substituted 1,2,3-triazole TSAO derivatives proved to be potent inhibitors of HIV-1 replication with higher antiviral selectivity than that of the parent TSAO prototype. PMID- 9865387 TI - Oxysterols, but not cholesterol, inhibit human immunodeficiency virus replication in vitro. AB - Oxysterols, oxygenated derivatives of cholesterol selected for their cytostatic activity and their inhibitory effect on cholesterol synthesis, have been investigated for their anti-human immunodeficiency virus (HIV) activity in vitro. The three oxysterols tested, 7 beta-hydroxycholesterol (7 beta-OHC), 25 hydroxycholesterol (25-OHC) and 7 beta, 25-dihydroxycholesterol (7,25-OHC), inhibit viral replication at micromolar concentrations. The selectivity indexes for 7 beta-OHC and 25-OHC are quite modest (2 to 8) but reproducible; the dihydroxycholesterol 7,25-OHC exhibited antiviral properties at concentrations 13 to 25-fold lower than the highest concentration tested at which no toxicity was measurable. Oxysterols are naturally occurring compounds, and we speculate on their physiological relevance in HIV-infected individuals. PMID- 9865388 TI - Synthesis and anti-human immunodeficiency virus type 1 integrase activity of hydroxybenzoic and hydroxycinnamic acid flavon-3-yl esters. AB - A series of new hydroxybenzoic and hydroxycinnamic acid flavon-3-yl esters were synthesized in order to obtain compounds targeting the human immunodeficiency virus (HIV) type 1 integrase (IN). The esters were tested for anti-IN and anti reverse transcriptase (RT) activity in enzyme assays and for anti-HIV-1, anti proliferative and anti-topoisomerase activity in cell-based assays. In enzyme assays, the two gallic acid flavon-3-yl esters showed a notable IN inhibition (IC50 values were 8.3 and 9.1 microM, respectively), while the two caffeic acid flavon-3-yl esters exhibited a modest activity (IC50 75 and 60 microM, respectively). Replacement of hydroxyl groups resulted in loss of potency. Caffeic acid 3',4'-dichloroflavon-3-yl ester also inhibited the RT activity whereas it was not active on human topoisomerases. It therefore represents an interesting example of a compound specifically targeting more than one step of the virus replication cycle. PMID- 9865389 TI - Anti-picornavirus activity of synthetic flavon-3-yl esters. AB - The in vitro antiviral activity against picornaviruses (rhinovirus serotype 1B and 14, and poliovirus type 2) of new synthetic 3-hydroxyflavones, 3 acetoxyflavones, and substituted cinnamic and benzoic acid flavon-3-yl esters was evaluated. The maximum non-toxic concentration of compounds was determined in a human cell line (HeLa) suitable for the replication of the three viruses. Their antiviral potency was measured by a plaque reduction assay. Generally, rhinoviruses exhibited a higher sensitivity to the new flavonoids than poliovirus. Flavones, with sterically small substituents in position 3, showed good activity against both rhinoviruses tested. However, the introduction of bulky substituents in the same position resulted in analogues with a higher toxicity and often with a lower efficacy. PMID- 9865390 TI - Warm up. PMID- 9865391 TI - Circus animals: desertion? PMID- 9865392 TI - Sport for people with a disability: the current state of play. PMID- 9865393 TI - Lightning strikes: danger overhead. PMID- 9865394 TI - Evidence based medicine in the musculoskeletal examination. PMID- 9865395 TI - Use of population attributable risk (PAR) in understanding the health benefits of physical activity. PMID- 9865396 TI - Viruses and athletes. PMID- 9865397 TI - Post-traumatic ectopic calcification in the muscles of athletes: a review. PMID- 9865400 TI - At what age should a child begin to undertake regular continuous exercise at moderate or high intensity? PMID- 9865399 TI - To count or not to count sports injuries? What is the question? PMID- 9865398 TI - Sports related injuries in Scottish adolescents aged 11-15. AB - OBJECTIVES: To measure the age and sex distribution of self reported sports and leisure injuries in a 12 month retrospective recall period among a representative national sample of Scottish adolescents, and to examine the characteristics (gender, age, handedness, and level of sports participation) of sports related injuries in relation to injuries sustained during other activities. DESIGN/SETTING: Self completion questionnaire survey administered in schools during April-June 1994. SUBJECTS: 4710 pupils aged 11, 13, and 15 years drawn from a representative sample of 270 classes with returns from 224 classes (83% completion rate). RESULTS: 42.1% of the sample reported a medically attended injury. These were significantly more frequent among boys but there were no significant age differences in overall frequency of injury. Sport related injuries accounted for 32.2% of all medically attended injuries. As with all injuries, frequency was greater in boys than girls at all ages, and there are differences in the pattern of lesions in sports and nonsports injuries. Lower limb injuries were more frequent than upper limb injury in sports (57.6 v 23.9%), whereas there were no differences in non-sports injuries (31.5 v 31.3%). Age and handedness differences in sports injury rates were also identified. High sports participation was significantly associated with a higher risk of injury in general and sports related injury in particular. CONCLUSION: Age, gender, handedness, and level of sports participation have been shown to be implicated in differential risk of sports related injury. Reducing sports injury among adolescents should be a priority, but research into the injury profiles of different sports is needed before detailed injury prevention strategies can be developed. PMID- 9865401 TI - Occurrence and trends in ski injuries in Norway. AB - OBJECTIVES: To provide an overview of ski injuries in Norway and to describe the changes between 1990 and 1996. METHODS: All ski injuries (7966) treated at four hospitals providing health care to a defined population of about 11% of the Norwegian population were registered prospectively from 1990 to 1996. For this study, 6462 injuries sustained in cross country skiing, downhill skiing, telemark skiing, and snowboarding were selected for further analysis. RESULTS: The relative distribution of ski injuries by type of skiing changed significantly from 1990 to 1996 (p<0.001). Injuries from snowboarding showed the highest relative increase, and those sustained during downhill skiing showed a decline. The proportion of fractures in all types of skiing increased during the study period (p=0.001). The proportion of injuries to knee/lower leg decreased and the proportion to the forearm/wrist/hand increased during the study period (p=0.03). The mean age of the injured skiers differed significantly among the different types of skiing activity (p<0.001): cross country skiers were the oldest followed by telemark skiers, downhill skiers, and snowboarders. CONCLUSIONS: Ski injury surveillance results in early detection of changes in temporal injury trends, allowing timely adjustment of injury prevention strategies. Injuries from snowboarding are on the increase in Norway, warranting more effective injury prevention measures. PMID- 9865402 TI - Renal and liver functions and muscle injuries during training and after competition in Thai boxers. AB - OBJECTIVE: To observe whether there are any injuries to muscle and deleterious effects on the liver and kidneys during training and after competition in Thai boxers. METHODS: Serum levels of intracellular enzymes and specific markers in the urine were measured during training and after fighting in Thai boxers. RESULTS: During the training period, the activities of muscle enzymes were significantly increased whereas those of the liver enzymes and creatinine clearance were not changed. After a match, on the other hand, both liver and muscle enzyme activities were elevated but renal function was decreased. CONCLUSIONS: The training protocol for Thai boxers has virtually no deleterious effect on liver and renal function, but damage to skeletal muscle cells may occur. However, competition may cause muscle injury without any obvious damage to the liver and kidneys. PMID- 9865403 TI - Isokinetic strength testing does not predict hamstring injury in Australian Rules footballers. AB - OBJECTIVE: To determine the relation of hamstring and quadriceps muscle strength and imbalance to hamstring injury using a prospective observational cohort study METHOD: A total of 102 senior male Australian Rules footballers aged 22.2 (3.6) years were tested at the start of a football season. Maximum voluntary concentric and eccentric torque of the hamstring and quadriceps muscles of both legs was assessed using a Kin-Com isokinetic dynamometer at angular velocities of 60 and 180 degrees/second. Twelve (11.8%) players sustained clinically diagnosed hamstring strains which caused them to miss one or more matches over the ensuing season. RESULTS: There were no significant differences for any of the isokinetic variables comparing the injured and non-injured legs in players with unilateral hamstring strains (n=9). Neither the injured nor the non-injured leg of injured players differed from the mean of left and right legs in non-injured players for any isokinetic variable. The hamstring to opposite hamstring ratios also did not differ between injured and non-injured players. A hamstring to opposite hamstring ratio of less than 0.90 and a hamstring to quadriceps ratio of less than 0.60 were not associated with an increased risk of hamstring injury. A significantly greater percentage of players who sustained a hamstring strain reported a history of hamstring strain compared with non-injured players (p=0.02). However, this was not related to muscle weakness or imbalance. CONCLUSIONS: Isokinetic muscle strength testing was not able to directly discriminate Australian Rules football players at risk for a hamstring injury. PMID- 9865404 TI - Bone mineral density and body composition of the United States Olympic women's field hockey team. AB - OBJECTIVE: To evaluate total bone mineral density (BMD) and body composition (% fat) in world class women field hockey players, members of the 1996 United States Olympic team. METHODS: Whole body BMD (g/cm2) and relative body fatness (% fat) were assessed by dual energy x ray absorptiometry using a Lunar DPX-L unit with software version 1.3z. Body composition was also estimated by hydrostatic weighing and the sum of seven skinfolds. RESULTS: Mean (SD) BMD was 1.253 (0.048) g/cm2 which is 113.2 (4.0)% of age and weight adjusted norms. Estimates of body composition from the three methods were similar (statistically non-significant): 16.1 (4.4)% fat from dual energy x ray absorptiometry, 17.6 (3.2)% from hydrostatic weighing, and 16.9 (2.6)% from the sum of seven skinfolds. Mean fat free mass was approximately 50 kg. CONCLUSIONS: The mean whole body BMD value for members of the 1996 United States Olympic women's field hockey team is one of the highest reported for any women's sports team. Moreover, the mean fat free mass per unit height was quite high and % fat was low. In this group of world class sportswomen, low % fat was not associated with low BMD. PMID- 9865405 TI - The New Zealand Rugby Injury and Performance Project: V. Epidemiology of a season of rugby injury. AB - OBJECTIVE: To describe the incidence, nature, and circumstances of injury experienced by a cohort of rugby union players during a full competitive club season. METHODS: A prospective cohort study followed up 356 male and female rugby players throughout the 1993 competitive club season. Players were interviewed by telephone each week to obtain information on the amount of rugby played and the injury experienced. RESULTS: Detailed information was collected for 4403 player games and 8653 player-practices. A total of 671 injury events were reported, of which 569 were rugby related. The injury rate for games was higher than that for practices (rate ratio 8.3). At 10.9 injuries per 100 player-games, males had a higher rate of injury than females at 6.1 injuries per 100 player-games (p<0.001). Injury rates varied by position, with male locks (13.0 injuries per 100 player-games) and female inside backs (12.3 injuries per 100 player-games) having the highest rate in their respective sexes. The lower limb was the body region most often injured in games (42.5%) and practices (58.4%). Sprains/strains were the most common type of injury in games (46.7%) and practices (76.1%). In games the tackle was the phase of play in which the most injuries occurred (40%), followed by rucks (17%) and mauls (12%). Thirteen per cent of game injury events were the result of foul play. CONCLUSIONS: Rugby injury was common among the study subjects and varied according to grade and gender. Identifying the causes of injuries in the tackle, lower limb injuries, and dealing with the issue of foul play are priority areas for the prevention of rugby injury. PMID- 9865406 TI - An examination of the frequency and severity of injuries and incidents at three levels of professional football. AB - OBJECTIVE: To assess the risk of injury to professional footballers during European international and English Premier and First Division league matches. METHODS: Videotaped recordings of 29, 49, and 93 matches from the 1996 European Championship, 1996/1997 English Premier season and 1994 to 1997 English First Division seasons respectively were analysed. During each match, several relevant variables, including the number of fouls, injuries, time of incident, player identity, and injury mechanism, were recorded. RESULTS: Significantly more free kicks were awarded during international matches than during league matches; however, there were no significant differences between the numbers of free kicks awarded over the three First Division seasons assessed. Between 1.7 and 3.0% of fouls resulted in a player requiring treatment for injury, but only 15-28% of all injuries resulted from foul play. In all "non-foul" situations, in which injury resulted, at least 60% still involved player to player contact. No significant differences in injury frequency were observed between playing positions or match halves. CONCLUSIONS: The results equate to a total of 808 players per season from the estimated 2600 players in the four English professional football leagues sustaining a match injury that caused them to miss at least one game. The large number of underlying "non-injury" incidents is identified as the reason for this level of injury rather than a higher ratio of "injury" to "non-injury" incidents in professional football compared with other occupations. PMID- 9865407 TI - Lightning injuries during snowy conditions. AB - Skiers and other snow sports enthusiasts can become lightning casualties. Two such accidents are reported, one being fatal. There are fewer warning signals of impending lightning strikes in winter-like conditions. However, outdoor activists should be aware of at least two suspicious clues: the appearance of convective clouds, and the presence of graupel (snow pellets) during precipitation. PMID- 9865408 TI - Provision of physiotherapy at the Tsing Ma Bridge international marathon and 10 km race in Hong Kong. AB - This report examines the delivery of sports physiotherapy at the Tsing Ma Bridge marathon and 10 km race in Hong Kong. The incidence, type of injury encountered, and treatment modalities selected are examined. The report also highlights the flexibility required when planning the provision of such a service. PMID- 9865409 TI - Chronic subdural haematoma, an unusual injury from playing basketball. AB - Although the literature records instances of acute epidural haematoma and acute subdural haematoma related to playing basketball, there has not been a report of chronic subdural haematoma as a basketball injury. With the burgeoning interest in this sport in the United Kingdom and Europe, the possibility of this particular neurotrauma increases. Such an injury, along with the diagnosis and management of this often insidious lesion, is documented in this case report. PMID- 9865410 TI - Bilateral stress fractures of the ulnae in a Kendo (Japanese fencing) player. AB - A case of bilateral stress fractures of the ulnae in a Kendo (Japanese fencing) player is reported; the mechanism of the fractures, particularly the difference in the site of fracture between the right and left ulna, is discussed. PMID- 9865411 TI - Posterior dislocation of the hip while playing basketball. AB - Injuries in basketball are usually to the ankles and knees. Dislocation of the hip is usually associated with severe trauma--for example, road traffic accidents. A case is reported here in which a 22 year old club basketball player slipped on landing from a jump shot, forcing him into a side splits position from which he sustained a posterior dislocation of the hip resulting in a sciatic nerve palsy. PMID- 9865412 TI - Dysbaric osteonecrosis of the shoulder in a sport scuba diver. AB - The previously unreported occurrence of dysbaric osteonecrosis in an amateur sport scuba diver who had no other identified cause of avascular bone necrosis is described. PMID- 9865414 TI - Poisoning with tilidine and naloxone: toxicokinetic and clinical observations. AB - The opioid analgesic tilidine and its metabolites were detected by high performance liquid chromatography (HPLC) with UV-detection in serum from a 28 year-old woman who ingested 100 ml Valoron (o)N containing 6.94 g of tilidine and about 0.56 g of naloxone with suicidal intention. Data on the toxicokinetics of tilidine in severe poisonings are missing. Therefore we followed serum concentrations of tilidine and metabolites for 48 or 96 h and for the first time calculated basic kinetic parameters in a massive life threatening poisoning. Serum concentration of tilidine 3 h after ingestion was 38.1 mg/l which is about 70 times of the upper therapeutic level in man and about ninefold above toxic concentrations known so far. The concentration of nortilidine, the primary active metabolite at this time was 18.8 mg/l. The terminal elimination half life's of tilidine and nortilidine were explicit, prolonged with 23.9 and 13.9 h respectively. The competitive opiate antagonist naloxone, which is added as a part of the industrially produced preparation Valoron N solution to minimise oral abuse is not able to prevent ventilatory depression in massive overdoses. PMID- 9865413 TI - Patellar tendinopathy: some aspects of basic science and clinical management. PMID- 9865415 TI - Oestrogenic activity of burgen bread to female rats. PMID- 9865416 TI - Functional responses of human neutrophils to sodium sulfite (Na2SO3) in vitro. AB - An influx of neutrophils into the airways is a common feature observed during pulmonary inflammation induced by air pollutants, including sulfur dioxide and sulfates. In the present study focusing on the in vitro interactions of sodium sulfite (Na2SO3) with human neutrophils, we confirm results indicating that this sulfite induces superoxide production (O2-) by itself. We demonstrated that this response can occur more rapidly than previously reported (within 5 min), and that Na2SO3 can act as a priming agent, in a concentration-dependent fashion, to the bacterial tripeptide N-formyl-methionine-leucine-phenylalanine (fMLP) by increasing O2-production. In addition, our results show that Na2SO3 induces gene expression in human neutrophils in a concentration-dependent manner as assessed by incorporation of 5-[3H] uridine into total RNA. However, it does not induce cell shape changes. We also demonstrated that Na2SO3 does not modulate neutrophil apoptosis nor reverse the well-known delaying effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on apoptosis. We conclude that Na2SO3 acts rapidly on neutrophil physiology, within a few minutes with respect to superoxide production, and a few hours (4 h) with respect to gene expression without altering a biological process such as the rate of apoptosis evaluated after a long period of incubation (20 h). We further conclude that Na2SO3-induced production of O2does not drive neutrophils to undergo apoptosis, a mechanism known to occur in other conditions. Therefore, the potential toxicity of Na2SO3 during pulmonary inflammation or lung-associated diseases may be related to its ability to induce superoxide production without altering neutrophil apoptosis rate. PMID- 9865418 TI - The use of radiopharmaceuticals as an effective toxicologic technique for studying nephrotoxicity of drugs: cyclosporine-A. AB - The concept of altered biologic behavior of administered radiopharmaceuticals is used routinely in clinical nuclear medicine to increase the sensitivity of diagnosis, monitor the efficacy of chemotherapeutic drugs and radiation treatment, and determine injury caused by a drug whose effect has exceeded its therapeutic value. In this study, cyclosporine-A (CsA) an immunosuppressant drug known to cause nephrotoxicity due to tubular impairment and Tc-99m MAG-3, a renal imaging radiopharmaceutical secreted by the tubules have been used in animal models to establish a method for investigating the nephrotoxicity of drugs. New Zealand rabbits and Wistar rats were used. The rabbits and rats were treated with 30 mg/kg of CsA for 4 and 28 consecutive days respectively. Plasma creatinine and urea were measured and renogram studies were performed in the rabbits prior to and on 1, 4, 8, 11 and 15 days after treatment with CsA. For the renogram, the rabbits were given an intravenous bolus injection of 44.4 MBq (1.5 mCi) of Tc-99m MAG-3. The Tmax, T1/2, TTHM and uptake slope of the Tc-99m MAG-3 were calculated. Each rat was injected intravenously with 185 MBq (5 mCi) of Tc-99m MAG-3, killed 3 min later, the kidneys removed and 20 mm frozen sections made. Autoradiograms were generated from the frozen sections. Creatinine and urea levels were also measured in the rats. There was no consistent difference in creatinine and urea levels between control and CsA treated rabbits and rats. However, for the rabbit, on day 1 or 4 after treatment, there was significant increase in the values of Tmax, T1/2, TTHM and uptake slope between the control and CsA treated animals, indicating intrarenal vasoconstriction and delayed transit of Tc-99m MAG-3 from the parenchyma to the collecting system. This delay is dramatically shown in the tissue autoradiograms of the rats. The results are consistent with reported nephrotoxicity of CsA using other techniques. The results of this study, therefore, indicate that the concept of altered biologic behavior of Tc-99m MAG-3 can be used effectively as a toxicologic method for studying nephrotoxicity of drugs as exemplified by CsA. PMID- 9865417 TI - Diisopropylglutathione ester protects A549 cells from the cytotoxic effects of sulphur mustard. AB - The A549 cell line was used to assess the ability of diisopropylglutathione (DIPE) to protect against a 100 microM challenge dose of sulphur mustard (HD) using gentian violet (GV), thiazolyl blue (MTT) and neutral red (NR) assays as indicators of cell culture viability. As part of a continuing study of the efficacy of protective nucleophiles as candidate treatments for HD poisoning, several different combinations of protectant and HD were used to determine the optimal means of protecting A549 cells from the effects of HD. It was found that DIPE (4 mM) could protect cells against the effects of HD though for optimal effect, DIPE had to be present at the time of HD challenge. Cultures protected with DIPE were up to 2.9-fold more viable than HD exposed cells 48 h after HD challenge when using the GV, MTT and NR assays to assess viability. Observations by phase contrast microscopy of GV stained cultures confirmed these findings. Pretreating A549 cultures with DIPE for 1 h followed by its removal prior to HD challenge did maintain cell viability, though at a relatively low level (only up to 1.4-fold more viable than HD only exposed cells). DIPE was also able to protect HD exposed A549 cultures when added to cell cultures at intervals of up to 12 to 15 min after the initial HD exposure, though viability tended to decrease over this period, so that at 1 h, addition of DIPE did not maintain the viability of the cultures. This is the first such report of the anti-HD protectant properties of DIPE in A549 cells. It is concluded that the protection observed against HD is probably largely due to extracellular inactivation of HD by DIPE. PMID- 9865420 TI - Chromosomes in the assessment of the effects of low levels of genotoxic agents. Leonard Sagan BELLE Award. Biological Effects of Low Level Exposure. PMID- 9865419 TI - Aspirin and heparin prevent hepatic blood stasis and thrombosis induced by the toxic glycoprotein Bolesatine in mice. AB - Bolesatine is a toxic glycoprotein isolated from Boletus satanas Lenz, which inhibits protein synthesis in vivo and in vitro. The LD50 (24 h) is 1 mg /kg bw (i.p.), in mice and rats. When given i.p. to mice (0.1 - 1.0 mg/kg bw) bolesatine induced thrombi and blood stasis in the liver, 5 - 21 h after injection, and modifications of the number of blood corpuscles in peripheral blood. These effects were efficiently reversed by aspirin, ticlopidin and heparin (as attested by histology and electron microscopy) which however failed to prevent death in animals given lethal doses. Together, these results showed that the death of bolesatine poisoned animals given high doses, was rather due to a combination of thrombosis and other toxic effects. In addition, they suggest that these antithrombotic drugs may overcome cases of human poisoning, with low exposures of this boletus, showing a hypertension probably due to mechanical obstruction which resists normal therapy. PMID- 9865421 TI - Gadolinium chloride toxicity in the mouse. AB - 1. Groups of five male and five female CD-1 mice received a single intravenous injection of gadolinium chloride at dosages of 0 (saline control), 0.05, 0.1 and 0.2 mmol/ kg. All mice were necropsied 48 h post dose. 2. Plasma analysis showed increases in concentrations of lactate dehydrogenase (both sexes), aspartate aminotransferase and alanine aminotransferase (females only) in the 0.2 mmol/kg group. Cholesterol was elevated at all dosages in both sexes whilst globulin was raised in both sexes at 0.1 and 0.2 mmol/kg. 3. Histological lesions were present at all dosages and increased in severity in a dose-related fashion. The most common lesions were: mineral emboli in capillaries, accumulation of mineral in the mononuclear phagocytic system, hepatocellular necrosis, and lymphoid depletion, necrosis and mineralisation in the spleen. 4. Such observations are similar to those in rats given gadolinium chloride and should be assessed when evaluating the toxicological profile of gadolinium containing compounds being developed for nuclear magnetic resonance imaging. PMID- 9865422 TI - Alterations in rat erythrocyte membrane due to hexachlorocyclohexane (technical) exposure. AB - 1. Hexachlorocyclohexane (HCH), an organochlorine pesticide having hydrophobic molecule is known to act on membranes. HCH mediated alterations in erythrocyte membrane occur through disorganization of the lipid bilayer. Therefore the changes in erythrocyte membrane fluidity, osmotic fragility and certain membrane bound enzymes were studied. Administration of HCH (technical) to rats at 5 mg/kg, orally, 5 days a week for 1, 2 and 3 months caused marked increase in erythrocyte membrane fluidity, osmotic fragility and decrease in levels of Na+, K(+)-ATPase, acetylcholinesterase in erythrocytes and glutathione in blood. 2. These changes indicate that HCH adversely affects membrane structure and function. PMID- 9865423 TI - Relationships of serum androgens and estrogens to prostate cancer risk: results from a prospective study in Finland. AB - Several lines of evidence suggest that sex hormones may be involved in the etiology of prostate cancer. We conducted a prospective nested case-control study to evaluate the relationships of serum androgens and estrogens to prostate cancer using serum collected at baseline for the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study. The 29,133 male smokers who participated in the trial were 50 69 years old at baseline. During 5-8 years of follow-up, 246 men were diagnosed with prostate cancer, and 116 of these were randomly selected for inclusion in the current study. For each case, two controls matched on age, date of blood collection, intervention group, and study center were selected. Hormones were measured in serum by RIA using standard procedures. None of the individual androgens or estrogens was significantly related to prostate cancer. These findings were unaltered by simultaneous evaluation of serum androgen and estrogen concentrations in multivariate models. These results do not support a strong relationship of serum androgens and estrogens with prostate cancer in smokers. Within-person variation in concentrations of some hormones may have contributed to the lack of significant associations. PMID- 9865424 TI - Debrisoquine hydroxylase (CYP2D6) and prostate cancer. AB - The p450 hepatic microsomal enzyme system metabolizes exogenous drugs and carcinogens. Debrisoquine hydroxylase (CYP2D6), one member of the p450 hemoproteins, has polymorphic expression leading to poor metabolism of debrisoquine and similar compounds in approximately 7% of Caucasians. The genetic locus for this enzyme has been characterized, and the mutations responsible for the slowed metabolism have been identified. Epidemiological studies of the CYP2D6 phenotype suggest an association between the normal or rapid metabolism phenotype and increased risk of lung and bladder cancer. Preliminary data have also suggested an association with prostate cancer (CaP). We used a PCR-based assay to investigate possible associations between the CYP2D6 B allele, the most common genetic mutation responsible for the poor metabolism phenotype, and CaP. Using genomic DNA isolated from peripheral blood, we genetically typed 571 men with CaP and 767 matched controls, all participants in the Physician's Health Study. Relative to men homozygous for the wild-type allele, heterozygotes for the B allele have an odds ratio of 1.19 (95% confidence interval, 0.94-1.51) for CaP, and men homozygous for the B allele have an odds ratio of 1.37 (95% confidence interval, 0.86-2.20). When analyzed as a trend over zero, one, or two copies of the B allele, there emerges a possible association between the B allele and an increased risk of CaP of borderline statistical significance (P = 0.07). PMID- 9865425 TI - NAT2, GSTM-1, cigarette smoking, and risk of colon cancer. AB - Cigarette smoking has been associated inconsistently with colon cancer. The extent to which genetic profile influences susceptibility to the inducement of colon cancer by cigarette smoking is not known. In this study, we evaluated the associations between smoking cigarettes and polymorphisms of the NAT2 and GSTM-1 genes using data obtained from an incident case-control study of 1993 cases of colon cancer and 2410 age- and sex- matched controls. Neither NAT2 nor GSTM-1 polymorphisms were significantly associated with colon cancer, except among older women, in whom the intermediate/rapid imputed phenotype was associated with increased risk of colon cancer [odds ratio (OR) = 1.4, 95% confidence interval (CI) = 1.0-1.81. Using several indicators of cigarette smoking, we observed no significant interaction between these genotypes and cigarette smoking and colon cancer. The major variation in association with colon cancer was from the amount of cigarette exposure, with those smoking a pack or more of cigarettes per day being at an approximately 40% increased risk of colon cancer; this association did not vary by genotype. However, those who stopped smoking 5-14 years prior to diagnosis and who where intermediate/rapid acetylators were at a slightly greater risk than those who were slow acetylators (for men, OR = 1.6, 95% CI = 1.0-2.4; for women, OR = 2.5, 95% CI = 1.4-4.4). Associations were similar when proximal and distal tumors were examined and separated for age at the time of diagnosis. The lack of an association does not rule out the possibility of other genetic polymorphisms interacting with cigarette smoke to cause colon cancer, nor does it take into account individual phenotypic variability. PMID- 9865426 TI - Effects of sodium selenite on deoxycholic acid-induced hyperproliferation of human colonic mucosa in short-term culture. AB - It has been shown that in vitro incubation of human colonic biopsies with the secondary bile acid deoxycholic acid (DCA) leads to the hyperproliferation of colonic crypt cells with an expansion of the proliferative zone, which is regarded as a biomarker of increased cancer risk. Sodium selenite (SSE), on the other hand, has been implicated as a protective agent in experimental studies, but toxic effects were reported as well, depending on the dose of SSE. To elucidate the effects of SSE on human colonic mucosa, biopsies from endoscopically normal sigmoid colon tissue of 30 subjects were incubated with 5 microM DCA or a combination of 5 microM DCA and SSE in concentrations of 5, 10, 20, 50, 80, and 100 microM, respectively. Equimolar NaCl incubations served as a control. Proliferating cells were labeled by bromodeoxyuridine immunohistochemistry, and the labeling index (LI) was computed. In the experiments using 5, 10, and 20 microM SSE, the whole crypt LI was significantly lower after DCA + SSE incubation (0.136, 0.118, and 0.110, respectively) compared to that after incubation with DCA alone (0.172, 0.157, and 0.165, respectively; P < 0.01). The corresponding LIs during DCA + SSE incubation were comparable to the LIs obtained after NaCl incubation (average LI = 0.14). Contrary to this finding, severe cell damage was observed in the biopsies that were incubated with the higher SSE concentrations of 50 microM and above. The antiproliferative effects of SSE may indicate a possible protective effect in the prevention of human colon cancer development. However, the observed toxic effects of higher SSE concentrations strongly suggest the need for additional studies before general recommendations for the use of SSE in colon cancer prevention can be made. PMID- 9865427 TI - Human metabolism and excretion of cancer chemoprotective glucosinolates and isothiocyanates of cruciferous vegetables. AB - Isothiocyanates and their naturally occurring glucosinolate precursors are widely consumed as part of a diet rich in cruciferous vegetables. When plant cells are damaged, glucosinolates are released and converted to isothiocyanates by the enzyme myrosinase. Many isothiocyanates inhibit the neoplastic effects of various carcinogens at a number of organ sites. Consequently, these agents are attracting attention as potential chemoprotectors against cancer. As a prerequisite to understanding the mechanism of the protective effects of these compounds, which is thought to involve the modulation of carcinogen metabolism by the induction of phase 2 detoxication enzymes and the inhibition of phase 1 carcinogen-activating enzymes, we examined the fate of ingested isothiocyanates and glucosinolates in humans. Recently developed novel methods for quantifying isothiocyanates (and glucosinolates after their quantitative conversion to isothiocyanates by purified myrosinase) and their urinary metabolites (largely dithiocarbamates) have made possible a detailed examination of the fates of isothiocyanates and glucosinolates of dietary crucifers. In a series of studies in normal volunteers, we made these findings. First, in nonsmokers, urinary dithiocarbamates were detected only after the consumption of cruciferous vegetables and condiments rich in isothiocyanates and/or glucosinolates. In sharp contrast, the consumption of noncrucifers (corn, tomatoes, green beans, and carrots) did not lead to the excretion of dithiocarbamates. Moreover, the quantities of dithiocarbamates excreted were related to the glucosinolate/isothiocyanate profiles of the cruciferous vegetables administered (kale, broccoli, green cabbage, and turnip roots). Second, eating prepared horseradish containing graded doses of isothiocyanates (12.3-74 micromol; mostly allyl isothiocyanate) led to a rapid excretion of proportionate amounts (42-44%) of urinary dithiocarbamates with first-order kinetics. The ingestion of broccoli in which myrosinase had been heat inactivated also led to proportionate but low (10-20%) recoveries of urinary dithiocarbamates. Broccoli samples subsequently treated with myrosinase to produce the cognate isothiocyanates were much more completely (47%) converted to dithiocarbamates. Finally, when bowel microflora were reduced by mechanical cleansing and antibiotics, the conversion of glucosinolates became negligible. These results establish that humans convert substantial amounts of isothiocyanates and glucosinolates to urinary dithiocarbamates that can be easily quantified, thus paving the way for meaningful studies of phase 2 enzyme induction in humans. PMID- 9865428 TI - Effects of soy isoflavones on estrogen and phytoestrogen metabolism in premenopausal women. AB - Isoflavones and lignans are soy phytoestrogens that have been suggested to be anticarcinogenic. The mechanisms by which they exert cancer-preventive effects may involve modulation of estrogen synthesis and metabolism. To evaluate this hypothesis, a randomized, cross-over soy isoflavone feeding study was performed in 12 healthy premenopausal women. The study consisted of three diet periods, each separated by a washout of approximately 3 weeks. Each diet period lasted for three menstrual cycles plus 9 days (averaging approximately 100 days), during which subjects consumed their habitual diets supplemented with soy protein powder providing 0.16 (control diet), 1.01, or 2.01 mg of total isoflavones per kg of body weight per day (10+/-1.1, 65+/-9.4, or 129+/-16 mg/day, respectively). A 72 h urine sample was collected during the midfollicular phase (days 7-9) of the fourth menstrual cycle in each diet period. Urine samples were analyzed for 10 phytoestrogens and 15 endogenous estrogens and their metabolites by a capillary gas chromatography-mass spectrometry method. Urinary excretion of isoflavonoids and lignans significantly increased with increased isoflavone consumption. Compared with the control diet, increased isoflavone consumption decreased urinary excretion of estradiol, estrone, estriol, and total estrogens, as well as excretion of the hypothesized genotoxic estrogen metabolites, 16alpha hydroxyestrone, 4-hydroxyestrone, and 4-hydroxyestradiol. Of importance are the observations of a significant increase in the 2-hydroxyestrone/16alpha hydroxyestrone ratio and a decrease in the genotoxic/total estrogens ratio. These data suggest that soy isoflavone consumption may exert cancer-preventive effects by decreasing estrogen synthesis and altering metabolism away from genotoxic metabolites toward inactive metabolites. PMID- 9865429 TI - Autoantibodies to the nuclear phosphoprotein nucleophosmin in breast cancer patients. AB - Nucleophosmin (NPM) is an estrogen-regulated nucleolar phosphoprotein; a substrate for phosphorylation by p34cdc2 kinase, protein kinase C, and casein kinase II; and a repressor of the transcriptional regulating activities of the YY1 and IFN regulatory factor-1 transcription factors. We have completed a pilot study to determine whether autoantibodies to NPM are present in breast cancer patients and explored the ability of these autoantibodies to predict recurrence in breast cancer patients. One hundred breast cancer patients were studied: 50 who recurred, and 50 matched for age and length of follow-up but who did not recur. Patients' sera were collected at the times of diagnosis (T1), six months before recurrence (T2), and at recurrence (T3). Recurrent and nonrecurrent patients did not differ in autoantibody levels at the times of diagnosis or recurrence. However, antiNPM autoantibody levels increase significantly between diagnosis and six months before recurrence in recurrent patients, whereas no change occurs over the comparable time period in nonrecurrent patients (repeated measures ANOVA; P = 0.041). At recurrence, the levels return to those seen at diagnosis. The greater the change in levels between T1 and T2, the greater the risk of recurrence within the next 6 months (conditional logistic regression: increase in risk for highest versus lowest tertile of change from T1 to T2; odds ratio, 3.25; 95% confidence interval, 1.04-10.18; P = 0.043). Consistent with the estrogenic/antiestrogenic regulation of the antigen in breast cancer cells, the levels of antiNPM autoantibodies are decreased 6 months before recurrence in patients treated with the antiestrogen tamoxifen (P = 0.012). The association between antiNPM levels and recurrence remained after adjustment for confounding factors. Further study of antiNPM autoantibody levels as a new and simple, intermediate serum biomarker for predicting both the timing of recurrence and monitoring response to endocrine manipulations in breast cancer patients is warranted. PMID- 9865430 TI - Epstein-Barr virus in Hodgkin's disease: correlation of risk factors and disease characteristics with molecular evidence of viral infection. AB - Risk factors suggestive of relatively late exposure to EBV have been consistently associated with Hodgkin's disease (HD) in younger adults. In addition, evidence of EBV infection has been found in the Reed-Sternberg cells themselves in about one-third to one-half of all HD cases. However, no study yet published has correlated these childhood social environment risk factors with the presence of EBV in Hodgkin's tumor cells. We examined whether EBV-positive HD occurs in those patients whose childhood environment would predispose them to relatively late exposure to EBV. The study population consisted of 102 cases of mixed cellularity (MC; n = 25) or nodular sclerosing (n = 77) HD. Samples that tested positive for either EBV-encoded RNA or latent membrane protein or both were considered EBV positive. Of the 102 cases, 83 completed a questionnaire regarding childhood social environment. The association with EBV-positivity was estimated by the odds ratio (OR) with 95% confidence intervals (CI). Twenty-two percent of the cases were EBV-positive. These cases were more likely to be MC (OR, 6.2; CI, 2.3-16.3) and male (OR, 3.4; CI, 1.3-9.0). History of infectious mononucleosis (IM) was not predictive of EBV-positivity, with only 3 of 14 such patients being EBV-positive (P = 0.82). Contrary to our hypothesis, no association between EBV and childhood environment risk factors was identified. The association of EBV with MC histology and male gender agrees with previous reports. The most intriguing finding was the dissociation between IM history and EBV-positivity, in that almost all of the cases with a history of IM were EBV-negative. PMID- 9865431 TI - Chromosome 8, occupational exposures, smoking, and acute nonlymphocytic leukemias: a population-based study. AB - In a previous epidemiological study on acute myelocytic leukemia (M. M. Crane et al., Cancer Epidemiol. Biomark. Prev., 5: 639-644, 1996), clonal aberrations in chromosome 8 have been reported to be in excess in smokers and in workers exposed to paints. In that study, cytogenetics was performed after therapy. In our report, we describe a population-based survey on nonlymphocytic leukemias in northern Italy, in which 79 patients (acute myelocytic leukemia, myelodysplastic syndromes, or other nonlymphocytic leukemias) were studied before cytotoxic therapy. We found 9 aberrations involving chromosome 8 (six +8, two -8, and one translocation), whereas abnormalities involving chromosomes 5 and 7 occurred with a low frequency compared with previous studies. Aberrations involving chromosome 8 were associated with smoking (odds ratio, 6.3; 95% confidence interval, 0.9 42.3; among smokers of 10 or more cigarettes/day: odds ratio, 14.2; 95% confidence interval, 1.4-142.3); +8 aberrations were found in 1 of 24 nonsmokers and in 5 of 38 smokers. Three +8 aberrations were found in 22 subjects potentially exposed to solvents or polycyclic aromatic hydrocarbons. The low frequency of chromosome 5 and 7 aberrations in our population-based series (compared with other studies) can be attributed to the recruitment before cytotoxic therapies. Aberrations involving chromosome 8 (particularly +8) were associated with smoking habits. Chromosome 8 includes the c-myc oncogene. PMID- 9865432 TI - Accuracy of DNA amplification from archival hematological slides for use in genetic biomarker studies. AB - Archival slides are a potentially useful source of DNA for mutation analyses in large population-based studies. However, it is unknown whether specimen age or histological stains alter the accuracy of Taq polymerase or induce secondary mutations in sample DNA. To address this question, we evaluated five methods for extraction of genomic DNA from archival bone marrow slides of 17 leukemia patients and analyzed exons 1 and 2 of the N- and K-ras genes for the presence of mutations. Of the five methods, optimal DNA purification was achieved by boiling and phenol:chloroform extraction. N-and K-ras exons 1 and 2 were independently amplified using 35 cycles of PCR, and 6-12 clones for each exon were isolated and individually sequenced for each patient. Mutations were confirmed by repeat extraction, cloning, and sequencing. Sixteen of 17 patient samples were successfully amplified (94%), including slides up to 29 years old. Twelve slides had been stained with Wright-Giemsa, I stained with toluidine blue, and 4 were unstained. A total of 16 single-base mutations were identified of 33,840 nucleotides sequenced. No insertions or deletions were identified. Six of 16 single-base mutations were previously described activating mutations in codon 13 of N-ras exon 1. The 10 other mutations were in other regions of the N- and K-ras genes and were not reproduced after repeat extraction, cloning, and sequencing. The frequency of these other alterations was I of 3384 bp. This value is comparable with the inherent error frequency for Taq polymerase. Our findings suggest that high fidelity DNA amplification can be achieved using archival hematological slides as old as 29 years and can be reliably used in genetic analyses. PMID- 9865433 TI - Mammographic densities and breast cancer risk. AB - The radiological appearance of the female breast varies among individuals because of differences in the relative amounts and X-ray attenuation characteristics of fat and epithelial and stromal tissues. Fat is radiolucent and appears dark on a mammogram, and epithelium and stroma are radiodense and appear light. We review here the evidence that these variations, known as mammographic parenchymal patterns, are related to risk of breast cancer. Studies that used quantitative measurement to classify mammographic patterns have consistently found that women with dense tissue in more than 60-75% of the breast are at four to six times greater risk of breast cancer than those with no densities. These risk estimates are independent of the effects of other risk factors and have been shown to persist over at least 10 years of follow up. Estimates of attributable risk suggest that this risk factor may account for as many as 30% of breast cancer cases. Mammographically dense breast tissue is associated both with epithelial proliferation and with stromal fibrosis. The relationship between these histological features and risk of breast cancer may by explained by the known actions of growth factors that are thought to play important roles in breast development and carcinogenesis. Mammographically dense tissue differs from most other breast cancer risk factors in the strength of the associated relative and attributable risks for breast cancer, and because it can be changed by hormonal and dietary interventions. This risk factor may be most useful as a means of investigating the etiology of breast cancer and of testing hypotheses about potential preventive strategies. PMID- 9865434 TI - Research issues in cancer survivorship: report of a workshop sponsored by the Office of Cancer Survivorship, National Cancer Institute. PMID- 9865435 TI - Correspondence re: C. Byrne et al., Predictors of dietary heterocyclic amine intake in three prospective cohorts. Cancer Epidemiol. Biomark. Prev., 7: 523 529, 1998. PMID- 9865437 TI - Laterally hypertrophic adenoids as a contributing factor in otitis media. AB - Adenoidectomy has been demonstrated in randomized controlled studies to be effective in the prevention of otitis media with effusion (OME). Despite an apparent lack of correlation between adenoid size and outcome, it has been suggested that the role of adenoid tissue in the pathophysiology of OME relates either to obstruction of the Eustachian tube (ET) with impairment of its function, or to adenoid tissue serving as a reservoir of infection. The aim of this study was to further delineate the relationship between adenoid hypertrophy and OME. This prospective survey was performed in an ambulatory care setting of a tertiary care pediatric hospital, where data was collected on 273 consecutive adenoidectomy patients. At the time of surgery, adenoid position in relation to the ET orifice was recorded as well as concurrent procedures performed (e.g. pressure equalization tubes; PET). Sixty percent of patients undergoing simultaneous PET insertion were found to have laterally hypertrophic adenoid tissue encroaching upon the ET orifice versus only 22% for those undergoing adenoidectomy alone. Thus, a strong correlation was found to exist between OME, requiring the placement of PET and lateral adenoids abutting the torus tubarius (chi2 = 39.12; P < 0.001). The clinical relevance of this finding is that it may allow the prediction of children with OME, who will benefit most from undergoing adenoidectomy. Patients with OME requiring PET insertion, who are found to have lateral adenoid hypertrophy encroaching upon the ET orifice, could be selected for adenoidectomy, thus allowing a more judicious application of this surgical procedure in the setting of OME. PMID- 9865436 TI - Distortion products otoacoustic emissions in diagnosis of hearing loss in Down syndrome. AB - This study was carried out to investigate the features of hearing impairment in subjects with Down syndrome. Forty-seven children and 14 adults with Down syndrome were included in the study. In all cases, a complete otorhinolaryngological examination was performed, followed by audiological assessment. Depending on age, intellectual level and middle ear status the following examinations were performed: pure-tone 'play audiometry', tympanometry, acoustic reflex, auditory brain response (ABR) and distortion products otoacoustic emissions (DPOAE). The results were compared with age matched control groups. Among the group of children with Down syndrome, we have frequently found impairment of the conductive function of the middle ear which was expressed by pathological tympanometry. Tympanometry of B and C type was detected in 56% of ears. The amplitude of DPOAE was lower in children with Down syndrome than in the control group. This difference was more expressed in adults with Down syndrome. DPOAE examination results in subjects with Down syndrome without conductive hearing loss indicate early age related inner ear impairment. PMID- 9865438 TI - Adenoidal and tonsillar lymphocyte subsets in AIDS children. AB - Tonsils and adenoids are secondary lymphoid organs exposed to the environment. The most important classifications of AIDS include the lymphocyte subsets of peripheral blood. We have studied the lymphocyte subsets in peripheral blood and secondary lymphoid organs in a control group of children suffering adenotonsillar pathology and in five children with AIDS and the same adenotonsillar pathology. The antigen surface markers were determined by flow cytometry in lymphocytes isolated from peripheral blood, and from tonsils and adenoids after tonsillectomy and adenoidectomy, in the control group and in children diagnosed with AIDS. The most important findings in tonsils and adenoids were a decrease of the total T lymphocytes, helper T lymphocytes and CD4/CD8 ratio; an increase of cytotoxic T lymphocytes and B lymphocytes, as well as a 200% increase in monocytes of AIDS affected children. These observations show the value of analyzing the lymphocyte subsets of the tonsils and adenoids of AIDS-affected children, and establishing an earlier relation to clinical symptoms. PMID- 9865439 TI - Wound management of the airway mucosa: comparison with skin in a rabbit model. AB - In comparison to the extensive study of skin wound healing, there have been few reports investigating mucosal wound healing. Our primary objective was to compare the natural progression of wound healing in airway mucosa to skin in a rabbit model. Split-thickness skin wounds and subglottic mucosal wounds created by drill injury were compared on days 0, 1, 3, 5, 7, 14 and 21 after injury. Histologic examination was performed by a veterinary pathologist blinded to sample identity. Subglottic wounds showed a 'fibrinous clot' overlying the epithelium, analogous to the fibrin crust in skin wounds. Re-epithelialization started on day 5 in the subglottic epithelium and was complete by day 14; fibroplasia and fibrosis in the lamina propria were present on days 7-21. This wound healing profile paralleled the skin epidermis and dermis, respectively. The epithelial changes, however, were temporally extended in the airway. Our secondary objective was to determine the effects of treating airway mucosa with a bioresorbable membrane, modified sodium hyaluronate and carboxymethylcellulose (modified HA/CMC), placed over the subglottic wounds of four rabbits after drill injury. Subglottic wounds treated with modified HA/CMC showed a more mature epithelium and less fibrosis on day 21. In this pilot study, the application of a bioresorbable membrane improved mucosal wound healing at both the epithelial and lamina propria levels. Clearly, a larger study must be performed to confirm this interesting observation. PMID- 9865440 TI - Congenital absence of the epiglottis. AB - The congenital absence of the epiglottis is a very rare anomaly. We present a patient who has a complete absence of the epiglottis, illustrated with an endoscopic photograph, review the literature and discuss the management of patients with this condition. PMID- 9865441 TI - Meningitis as a complication of tonsillectomy. AB - Adenotonsillar surgery remains among the most commonly-performed pediatric surgical procedures. The complication rate of tonsillectomy is generally considered to be in a range of 1-5%. Bacterial meningitis following adenotonsillectomy is a rarely reported complication. Bacterial meningitis is a potentially fatal acute infectious disease caused by a variety of micro organisms. Current case fatality rates associated with this entity can be as low as 2% in infants and children and as high as 30% in neonates. The successful management of meningitis depends on early clinical suspicion and diagnosis, with prompt medical treatment using high-dose, broad-spectrum antibiotics that adequately cross the blood-brain barrier. We present a case of meningitis complicating a tonsillectomy procedure, in a 7-year-old girl. The diagnosis and treatment of this rare complication is illustrated, and the possible etiology is discussed. PMID- 9865442 TI - Multicentric infantile myofibromatosis. AB - Infantile myofibromatosis (IM) is a rare tumor of infancy and childhood, typically presenting as a firm, nodular mass involving soft tissue, bone or viscera. Approximately one-third of cases involve the head and neck. These tumors can be solitary or multicentric. Biopsy reveals tumor cells that resemble myofibroblasts. Spontaneous regression may occur. A high degree of suspicion is necessary to differentiate this entity from other more aggressive processes histiocytosis, fibrosarcoma, rhabdomyosarcoma. We describe the case of a male infant with multicentric myofibromatosis, presenting with multiple thoraco abdominal subcutaneous nodules and lytic mass lesions of the temporal bone and calvarium. The characteristic clinical, radiologic and histopathologic features of this process are reviewed along with diagnostic and therapeutic options. PMID- 9865443 TI - Gastric cyst of the oral cavity. AB - A case of a heterotopic gastric cyst of the oral cavity is described and a review of the literature, with emphasis on the possible etiological theories of this rare lesion, is presented. This developmental lesion is found more commonly in males. It most likely arises from misplaced embryonal tissue. Surgical treatment is the preferred choice of treatment, and CO2 laser is a safe alternative surgical approach for the removal of this lesion. PMID- 9865444 TI - Mucoepidermoid carcinoma of the trachea in a child. AB - Primary tracheal neoplasms in children, especially malignant lesions, are extremely rare. We report a unique case of low-grade mucoepidermoid carcinoma of the trachea in a 4-year old child, presenting as new-onset asthma. Using the ventilating resectoscope, endoscopic excision was accomplished without complications. The case is presented with documentation including airway films, endoscopic photographs, and histopathology. The diagnosis of tracheal masses depends upon maintaining a high index of suspicion, complemented by imaging and timely diagnostic endoscopy. A review of the literature is included. PMID- 9865445 TI - Tracheobronchial mucoepidermoid carcinoma in childhood and adolescence: case report and review of the literature. AB - In children and adolescents, primary neoplasms of the tracheobronchial tree and lungs are rare, with most tumors involving the respiratory system being metastatic, small, blue cell tumors of childhood. Of the primary pulmonary neoplasms, most are malignant with mucoepidermoid carcinoma representing about 10% of these malignant tumors. We present an 8-year-old Hispanic male with hemoptysis and several episodes of pneumonia which initially was thought to be infectious upon biopsy during bronchoscopy, but proved to be mucoepidermoid carcinoma of the tracheobronchial tree by microscopic examination during an open lung biopsy. This rare tumor is more common in adults than in children, and infrequently presents with hemoptysis. Mucoepidermoid tumors of the tracheobronchial tree carry a more favorable prognosis in children than adults. In the adult population, the overall mortality is slightly less than 30%. In contrast, of the 31 reported cases of tracheobronchial mucoepidermoid carcinoma in pediatrics, all children are free of tumor involvement with a mean follow-up period of 5.8 years (range, 0.7-21 years). Based upon the available clinical outcome and survival data, it would appear that tracheobronchial mucoepidermoid carcinoma may be successfully managed by surgical intervention alone in children and adolescents. PMID- 9865446 TI - Inherited disorders of epidermal keratinization. AB - There have been a number of major discoveries recently in the field of dermatological science which have enabled us to determine the causes of inherited skin diseases of previously unknown etiology. In this paper we will review some important aspects of the biology of epidermal differentiation and the recent advances in understanding of the molecular mechanism underlying genetic diseases of keratinization. PMID- 9865447 TI - Differential effect of etretinate on proliferation and extracellular matrix metabolism of human dermal fibroblasts from elderly and young individuals in a novel three-dimensional culture. AB - The effect of etretinate on proliferation and biosynthesis of collagens and glycosaminoglycans (GAGs) were investigated using human dermal fibroblasts in a novel three-dimensional culture supplemented with L-ascorbic acid 2-phosphate. Fibroblasts from two young and two elderly individuals were studied at different concentrations of etretinate, 0.25, 1.0 and 2.5 microg/ml. Collagens (hydroxyproline) and GAGs (disaccharide units) extracted from the cell layer were analyzed and quantified biochemically by high-performance liquid chromatography (HPLC). Etretinate showed no significant effect on fibroblast proliferation either in the monolayer or the three-dimensional culture. Etretinate increased the collagen or GAGs content in the cell layer, which was prominent at etretinate concentrations of 1.0 and 2.5 or 0.25 microg/ml, respectively in fibroblasts from the elderly (P < 0.05). This effect was not seen in dermal fibroblast from the young. These results suggest that etretinate may have the differential effect on collagen and GAG metabolisms depending on the donor age of the cultured fibroblasts. PMID- 9865448 TI - Dry condition affects desquamation of stratum corneum in vivo. AB - We examined whether a dry condition actually induces scaly skin in vivo. Hairless mice were kept in a high humidity condition or a low humidity condition and skin changes were examined. Scales appeared on the backs of mice kept for 3 days under the dry condition. The weight of stratum corneum (SC) was increased at this point, and these alterations were not accompanied with hyperproliferation of the nucleated cell layer of the epidermis. A decrease of desmosomal degradation was observed, though, desquamation-related enzyme activity was not altered. The regulation mechanism of desquamation is not yet clear, however, in vitro experiments suggest that the water content in SC is an important factor. The water content of SC was decreased in the dry condition. These results indicate that a dry environment perturbs desmosome degradation in intact SC by decreasing the water content of SC, and the consequent impairment of desquamation in normal skin in vivo may lead to the induction of a scaly skin surface. PMID- 9865451 TI - Health effects of socioeconomic and geographic status. PMID- 9865449 TI - Increased numbers of CD68 antigen positive dendritic epidermal cells and upregulation of CLA (cutaneous lymphocyte-associated antigen) expression on these cells in various skin diseases. AB - CD68 is a myelomonocytic marker identified in human dermal macrophages. Although the existence of CD68 + dendritic epidermal cells has been reported, their characteristics have not been well elucidated. Cutaneous lymphocyte-associated antigen (CLA) is a homing receptor of cutaneous inflammatory T cells. Our recent report suggested that CLA was a homing molecule of CD1a+ Langerhans cells (LC) in the skin. In the present study we tested whether CD68 and CLA+ dendritic epidermal cells were present in skin specimens of normal skin and diseased skin such as lichen planus (LP), psoriasis vulgaris (PS), discoid lupus erythematosus (DLE), basal cell epithelioma (BCE), squamous cell carcinoma (SCC), irritated seborrheic keratosis (iSK), and Bowen's disease (BD). CD68+ dendritic epidermal cells were identified in normal skin and consisted of half the population of CD1a+ LC. These data indicate that CD68+ dendritic epidermal cells constitute a subpopulation of CD1a+ LC. CLA was expressed on a small percentage of CD68+ dendritic epidermal cells in normal skin. A remarkably increased number of CD68+ dendritic epidermal cells and upregulation of CLA on CD68+ dendritic epidermal cells were observed in diseased skin. The percentage of CLA+ cells among all CD68+ dendritic epidermal cells was less than that of CLA+ cells among all CD1a+ LC in diseased skin. The percentage of CLA+ cells among all CD68+ dendritic dermal cells was much less than that of CLA+ cells among all CD1a+ dendritic dermal cells. In normal skin, the epidermis showed minimal expression of monocyte chemoattractant protein (MCP)-1 and TGF-beta2, and no expression of TGF-beta1. In diseased skin, the epidermis showed elevated, but still moderate immunoreactivity for MCP-1. Slightly enhanced immunoreactivity for TGF-beta2, but not for TGF beta1, was observed in the epidermis of diseased skin. Increased epidermal MCP-1 immunohistochemical staining was associated with the increased number of CD68 dendritic epidermal cells. These data suggest the possibility that MCP-1 secretion from the epidermis can affect the migration of CD68; Cutaneous lymphocyte-associated antigen; Monocyte chemoattractant protein-1; TAF-beta. PMID- 9865450 TI - Assessment of in vitro percutaneous absorption of glycolic acid through human skin sections using a flow-through diffusion cell system. AB - The percutaneous absorption studies were performed using a flow-through diffusion cell system with skin specimens from 24 healthy women to assess the penetration of glycolic acid (GA). Percentages of GA, based on 14C-labelled activity, found in the skin after application of 4% GA at pH 2.0 or pH 3.8, after 24 h were as follows: stratum corneum (SC)= 2.65+/-1.80 versus 1.13+/-1.14 (P<0.05); viable skin (VS)= 13.46+/-7.44 versus 2.23+/-1.51 (P< 0.05) and effluent fraction (EF) = 12.22+/-9.03 versus 1.42+/-0.77 (P < 0.05), respectively. The applications of 4 60%, GA at their native pH resulted in an increased penetration of GA through the skin. For example, application of 20% GA, pH 1.9, resulted in the following values: SC = 2.69+/-2.26 (P > 0.05); VS = 4.88+/-4.05 (P > 0.05) and EF = 30.69+/ 13.25 (P < 0.05). Duration of application also affected the extent of penetration of drug. For example, application of 20% GA, pH 1.9, for 6 h resulted in the following levels: SC = 1.16+/-0.80 (P < 0.05); VS = 4.07+/-1.78 (P > 0.05) and EF = 6.12+/-4.95 (P < 0.05). IN CONCLUSION: (i) absorption of GA in human skin are pH-, strength- and time-dependent; and (ii) the in vitro method appears to provide an appropriate model to reflect in vivo absorption of GA through human skin. PMID- 9865453 TI - Clinical and prognostic significance of hyperfibrinogenemia in cerebral ischemia. AB - In order to evaluate the clinical and prognostic significance of early hyperfibrinogenemia in patients with transient ischemic attack (TIA) and ischemic cerebral infarction (ICI), we analyzed the relationships between plasma fibrinogen, brain damage severity, clinical status on admission and intra hospital mortality. Vascular damage severity was estimated by measuring the necrotic area by computed axial tomography (CT) and indirectly by means of changes in some plasma enzymes (CK, LDH, GPT/ALT, and GOT/AST). Plasma fibrinogen levels were statistically higher in ICI than in TIA and control subjects (p < 0.0005; analysis of variance). Moreover, plasma fibrinogen was directly related to the extension of the necrotic area at CT scan (p < 0.05) and in ICI patients was positively correlated with CK (r = 0.50, p < 0.01), LDH (r = 0.41, p < 0.05) and GOT/AST (r = 0.42, p < 0.05) serum levels, but not with GPT/ALT. A higher plasma fibrinogen value was observed in patients with stupor or coma compared with those with alert consciousness (p < 0.05). In patients who died during hospitalization, fibrinogen levels were higher than those of subjects who were discharged (p < 0.005). The results indicate that in the early phase of cerebral ischemia, plasma fibrinogen levels are related to the severity of the clinical status and to the extension of the brain vascular damage, thus representing a negative clinical and prognostic factor of the disease. PMID- 9865452 TI - Medical evaluation of Persian Gulf veterans with fatigue and/or chemical sensitivity. AB - The purpose of this study was to determine if Gulf War veterans with complaints of severe fatigue and/or chemical sensitivity (n = 72) fulfill case definitions for chronic fatigue syndrome (CFS) and/or multiple chemical sensitivity (MCS) and to compare the characteristics of those veterans who received a diagnosis of CFS (n = 24) to a group of non-veterans diagnosed with CFS (n = 95). Thirty-three veterans received a diagnosis of CFS with 14 having MCS concurrently; an additional six had MCS but did not fulfill a case definition for CFS. The group of fatigued veterans receiving a diagnosis of CFS was comprised of significantly fewer women and fewer Caucasians than the civilian group, and significantly fewer veterans reported a sudden onset to their illness. Veterans with CFS had a milder form of the illness than their civilian counterparts based on medical examiner assessment of the severity of the symptoms, reported days of reduced activity, and ability to work. Since CFS in veterans seems less severe than that seen in civilians, the prognosis for recovery of veterans with this disorder may be better. PMID- 9865454 TI - Decreased fecal bile acid output in patients with coronary atherosclerosis. AB - In most patients with atherosclerosis, the underlying metabolic derangement remains undefined. Animal experiments have suggested that the ability to produce and excrete large amounts of bile acids may be an adaptation mechanism to cholesterol overload protecting against the atherogenic effects of cholesterol. However, there are very few data on bile acid excretion in human atherosclerosis. In the present study, we have investigated fecal bile acid secretion in subjects with and without coronary artery disease. The target group consisted of 30 patients with proven coronary artery disease and the control group consisted of 27 matched subjects without clinical or laboratory evidence of coronary atherosclerosis. Fecal bile acids were measured by gas-liquid chromatography from 24-hr stool collections under a controlled diet. The patients excreted significantly less bile acids than the controls (325+/-135 vs. 592+/-223 mg/day, respectively, p < 0.0001). The difference was primarily due to a reduced excretion of secondary bile acids. Less than 50% of deoxycholate was excreted by patients (180+/-81 mg/day) as compared to controls (367+/-168 mg/day, p < 0.0002), while lithocholic acid excretion was 111+/-62 mg/day in patients vs. 190 +/-70 mg/day in controls (p < 0.005). The fecal output of the two primary bile acids, cholic and chenodeoxycholic acid, did not differ significantly between patients and controls. The fecal output of total bile acids correlated with that of both secondary bile acids in patients as well as in controls. These findings suggest that patients with coronary heart disease are unable to excrete adequate amounts of bile acids to rid themselves of excess cholesterol, even if they are able to maintain a plasma cholesterol level comparable to that of healthy controls. PMID- 9865455 TI - Effect of acute increase of interstitial myocardial fluid on ventricular function in isolated working rat hearts. AB - An acute increase of myocardial interstitial fluid may affect ventricular function. In the present study we evaluated the effects of acute changes of myocardial tissue fluid on cardiac function and ultrastructural morphometry. Isolated rat hearts were perfused for 100 min in the working heart mode. Hearts were distributed into 5 groups: controls [perfused with Krebs-Henseleit (KH) isotonic buffer to rat plasma, KH, 287 mOsm], moderate hyposmotic perfusion (75% Hyposm: perfusion with 75% diluted KH, 216 mOsm), highly hyposmotic perfusion (60% Hyposm: perfusion with 60% diluted KH, 170 mOsm), afterload increase (Pre over: isotonic perfused hearts subjected to an increase of afterload from 72 to 145 cm H2O) and ion dilution (Ion-dil: hearts perfused with a 60% KH with 115 mM sucrose, isotonic, 287 mOsm). We evaluated functional changes, markers of cellular necrosis or damage (CPK, LDH and purine release in coronary effluent), heart weight changes (weight gain and ww/dw ratio) and ultrastructural morphometry (analysis of cell damage, interstitial area, and mitochondrial alterations by a computerized image analysis system). The ww/dw ratio increased significantly only in 60% Hyposm (+140%, p < 0.001) and Pre-over (+63%, p < 0.001 vs control) groups. An impaired myocardial function in 60% Hyposm, Pre-over and Ion-dil groups was observed with cardiac failure at 50, 60 and 60 min, respectively. Enzyme release was significant higher in 60% Hyposm and Pre-over groups and was related to heart weight gain (r = 0.85, p < 0.001). Ultrastructural analysis confirmed a significant increase of interstitial space area (ISA) and mitochondrial damage in 60% Hyposm and Pre-over groups (p < 0.001); a significant (p < 0.05) increase was observed in the Ion-dil group; in 75% Hyposm group, a significant increase of mitochondrial damage was detected (p < 0.05). In brief, a higher functional and morphological deterioration was observed in hearts in which a more evident interstitial edema was detected (60% Hyposm and Pre-over groups). We conclude that, in the experimental condition, an acute increase of myocardial interstitial tissue fluid directly compromises left ventricular function and contributes to the ultrastructural damage to the myocardium. PMID- 9865456 TI - CD4+ and CD8+ lymphocyte and cortisol response patterns in elderly and young males after methylprednisolone exposure. AB - The elderly have impaired cellular immunity and are more predisposed to opportunistic infections after long term glucocorticoid treatment. No data, examining the response of lymphocyte subsets (CD4+, CD8+) under baseline conditions and after exposure to methylprednisolone in young and elderly males, are available. This crossover study examined lymphocyte subsets and cortisol response patterns in seven elderly males (66-82 years) and five young males (24 37 years) randomized into Phase I (24 hr baseline) and Phase II (10 mg intravenous dose of methylprednisolone). Whole blood samples were obtained at 0, 4, 8, 12 and 24 hr to determine total lymphocytes and CD4+ and CD8+ cells utilizing monoclonal antibodies and flow cytometry. The absolute number of lymphocyte subsets and the lymphocyte area under the time curve (AUC) were measured and a 12 and 24 hr lymphocyte response ratio (AUC Phase II divided by AUC Phase I) was determined. Serial plasma samples over 24 hours were collected to quantitate cortisol (Phase I) and methylprednisolone concurrent with cortisol (Phase II). Pharmacokinetic parameters were generated and the cortisol AUC was determined. The AUC values for lymphocytes and cortisol from Phase II quantitated the pharmacologic response to methylprednisolone exposure while Phase I data described the interpatient variability in these parameters. Diurnal patterns for lymphocytes and cortisol were noted in all subjects during Phase I. The mean CD4+ and CD8+ lymphocyte AUC from 0 to 24 hr during Phase I was significantly smaller for the elderly when compared to young men. However, after exposure to methylprednisolone, lymphopenia occurred in all subjects with a mean decline of 54% in the elderly and 60% (p = 0.44) in young subjects for the total lymphocyte count and returned to baseline by 8-12 hr. During Phase II, the CD4+ lymphocytes (72% decline in elderly; 70% in young; p = 0.71) demonstrated a more notable decline than CD8+ cells (44% decline in elderly; 52% in young; p = 0.31) with a nadir occurring between 4 to 6 hr for both subsets. The lymphocyte response ratio was not significantly different between groups for total, CD4+, and CD8+ cells at 12 hr or 24 hr determinations. A slower clearance of methylprednisolone was noted in the elderly (mean: 256 mL/hr/Kg) than in the young men (mean: 359 mL/hr/Kg; p < 0.05) during Phase II with no significant difference found between groups for volume of distribution, elimination rate constant or half-life. A significantly smaller cortisol suppression ratio [0.36+/-0.11 (elderly) versus 0.58+/-0.11 (young), p = 0.01] which indicates a more profound cortisol suppression was noted. A significant correlation of -0.61 (p < 0.05) between drug exposure (methylprednisolone AUC) and pharmacologic effect (cortisol suppression ratio) was noted for the combined data in the young and elderly males. During Phase I, the CD4+ and CD8+ lymphocyte AUC was significantly smaller in the elderly. A definite suppression pattern for total, CD4+ and CD8+ lymphocytes and cortisol was noted after methylprednisolone exposure in young and elderly males. An age dependent suppression of cortisol during Phase II was noted but the degree of lymphopenia after drug exposure did not differ between the young and elderly group for any of the cell subsets. These data from healthy elderly provide a basis for further studies to assess immunologic and endocrinologic responses among elderly patients requiring chronic glucocorticoid therapy. PMID- 9865457 TI - A rare case of a thyrotropin-secreting pituitary tumor which responded to methimazole. AB - A 52-year-old Japanese male complained of palpitations and excessive sweating. He showed evidence of hyperthyroidism, but without suppression of the serum thyroid stimulating hormone (TSH). On admission, the serum level of the alpha-subunit of TSH was elevated, but tests for thyroid autoantibodies were negative. The TSH response to thyrotropin-releasing hormone (TRH) was blunted. Imaging studies revealed a pituitary tumor 2 cm in diameter. Administration of methimazole lowered the serum levels of thyroid hormones, and elevated the serum level of TSH. These findings led to the diagnosis of a thyrotropin-secreting pituitary tumor. Immunohistochemical analysis and electron microscopic findings verified the diagnosis. PMID- 9865458 TI - Effect of streptococcal lyzate OK-432 on peripheral blood mononuclear cells in gastric cancer patients. AB - OK-432, a killed preparation of Streptococcus pyogenes, as well as Bacillus Calmette-Guerin (BCG) and Corynebacterium parvum are all known biological response modifiers. To examine the immunomodulatory effects of OK-432, natural killer cell activity and cytokine production by peripheral blood mononuclear cells (PBMCs) were assessed in 32 patients with gastric cancer. Skin tests for Streptococcus pyogenes A-3Su (Su-PS) and BCG were performed in all patients. Other nutritional and immunological parameters were also determined. OK-432 treated PBMCs showed a significant increase of cytotoxicity against K562 cells (p < 0.01). Increased levels of interferon-gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha) and interleukin-2 (IL-2) were found in the supernatants of cultures treated with OK-432 in 29 (90.6%), 20 (62.5%), and 8 (25.0%) out of 32 patients, respectively. Natural killer cell activity, IFN-gamma production, and the Su-PS skin test were positively correlated (p < 0.01). In contrast, the BCG test and other markers were not correlated with natural killer cell activity and IFN-gamma production. These results suggest that the Su-PS skin test could predict OK-432-induced natural killer cell activity and IFN-gamma production in patients with gastric cancer, and was therefore useful to determine whether patients were responders to OK-432. PMID- 9865459 TI - Neutrophilia of infection/inflammation: are we really dealing with "inflamed" leukocytes? AB - We adopted whole blood flow cytometry and direct labeling of the CD11b/CD18 and CD62L antigens to study the relationship between their expression and leukocytosis in patients with infection/inflammation, acute stress and healthy volunteers. Mean +/- S.D. channel fluorescence intensity of CD11b/CD18 antigen on peripheral blood polymorphonuclears did not differ between patients with infection/ inflammation (173+/-78) and controls (167+/-72), but was significantly (p = 0.04) reduced in stress (135+/-60). No correlation was found between CD11b/CD18 antigen level and either polymorphonuclears absolute number or serum C reactive protein. A significant negative correlation was noted between CD62L antigen expression on polymorphonuclears and their absolute number. We assume that cells with increased CD11b/CD18 surface concentrations are retained in the capillaries and that part of the leukocytes in the peripheral blood are stressed leukocytes with reduced CD11b/CD18. Thus, leukocytes detected in peripheral blood are not necessarily the most "inflamed" ones. PMID- 9865460 TI - Detection of left ventricular enlargement by electrocariography. AB - Cardiomegaly is one of the commonest findings encountered in daily clinical practice, and its differential diagnosis is a common clinical problem. There are many electrocardiological (ECG) criteria known for left ventricular hypertrophy (LVH), but its limitations have also been suggested. We evaluated 102 patients fulfilling the ECG criteria of precordial and limb lead for LVH with echocardiographic findings as a gold standard. Among these 102 patients, the echocardiogram revealed 38 subjects with LVH, 26 subjects with left ventricular dilatation (LVD), 7 subjects with both findings, and 31 subjects with neither findings. Precordial criteria such as SV1+RV5 or RV6 > 30 mm, SV1 or SV2+RV5 > 35 mm, R+S > 40 mm, SV1 or SV2+RV5 or RV6 > 35 mm, SV2+RV4 or RV5 > 35 mm, high in sensitivity and low in specificity for LVD and LVH, are appropriate for screening LVD and LVH. Cornell limb lead criterion, SV3+RaVL > 28 mm (male), SV3+RaVL > 20 mm (female), high in sensitivity and specificity only for LVH, is the best elecrocardiographic criterion to evaluate LVH. Precordial and limb lead criteria such as R> 13 mm, RaVL > 12 mm, RaVF > 20 mm, onset of intrinsicoid deflection in V5 or V6> 0.05 sec, left axis deviation -30 degrees to -90 degrees, low in sensitivity, and high in specificity, are useful to rule out LVH and/or LVD. Our findings suggest LVD and LVH can be evaluated by ECG, but similar sensitivity and specificity for both LVH and LVD makes separation of LVH from LVD unattainable. PMID- 9865461 TI - Intussuception as a complication of chronic lymphocytic leukemia. AB - The case of a 58-year-old man with chronic lymphocytic leukemia (B-cell type) who later developed an intussuception of the small intestine due to a tumor is described. The histopathological findings of the removed tumor were compatible with those of diffuse small lymphocytic lymphoma (B-cell type). The residual tumor became smaller with CHOP therapy. It is considered that CLL cell infiltration into the small intestine resulted in intussuception. Since many tumors and lymphomas can form polypoid lesions causing an intussuception. This is a possible complication of CLL and it could occur even when the WBC count is well controlled. PMID- 9865463 TI - Kinetics of platelet P-selectin mobilization: concurrent surface expression and release induced by thrombin or PMA, and inhibition by the NO donor SNAP. AB - Activated platelets and endothelium surface express the cell adhesion molecule P selectin (CD62P), which plays an important role in mediating interactions with leukocytes. Increased levels of a functional soluble form of P-selectin (sP selectin) have been reported in several pathological states but it is not clear whether this circulating sP-selectin originates from platelets and/or endothelial cells. Here we describe the concurrent kinetics of intracellular storage, surface expression and release of platelet P-selectin induced by thrombin or the protein kinase C activator PMA. Platelet activation with submaximal concentrations of thrombin (0.1 U/ml) resulted in a rapid decrease of intracellular P-selectin. This decrease of intracellular P-selectin concurred with a gradual increase of surface expression and an initial increase of sP-selectin. Our results indicate that intracellular stores of P-selectin were only partly mobilized upon activation with submaximal concentrations of thrombin. A high concentration of thrombin (1.0 U/ml) induced a rapid and nearly total decrease of intracellular stores and a more pronounced, but transient, increase of surface expression. The release of P-selectin was fast and occurred during the initial activation phase. The NO donor SNAP inhibited both surface expression and release of platelet P selectin in a similar manner. PMA (0.1-1.0 microM) mediated a more slow, gradual and sustained surface expression and release of P-selectin than thrombin. Thus, surface expression and release of platelet P-selectin show different kinetics depending on the mode of activation. PMID- 9865462 TI - Functional interaction between RhoB and the transcription factor DB1. AB - RhoB has been implicated in cell growth control, actin regulation, adhesion dependent viability, and gene expression, but its effector functions are poorly defined. Prenylation is important for the physiological functions of Rho proteins, so to identify RhoB effector functions we identified proteins whose interaction was sensitive to prenylation. Here we report the investigation of one such protein, an ubiquitously expressed transcription factor termed DB1 that was originally cloned as a Tax-activated regulator of the IL3 promoter. The RhoB binding domain in DB1 was located in a functionally undefined region upstream and separable from its zinc finger DNA binding domain. DB1 interacted strongly with prenylated RhoB but weakly with RhoA and not at all with H-Ras. Functional interaction was supported by the identification of prenylated species of RhoB in the nuclear membrane and in an intranuclear laminar region, where they were available for DB1 association in principle, and by the ability of RhoB to inhibit transcriptional activation by DB1, whereas RhoA or Ras had little or no effect, respectively. The results of this study suggest a novel mechanism by which certain Rho proteins may regulate transcription, through sequestration of a transcription factor. PMID- 9865464 TI - Physical, contractile and calcium handling properties of neonatal cardiac myocytes cultured on different matrices. AB - Extracellular matrix components play a vital role in the determination of heart cell growth, development of spontaneous contractile activity and morphologic differentiation. In this work we studied the physical and contractile changes in neonatal rat cardiac myocytes over the first four days of growth on three different extracellular matrices. We compared commercial laminin and fibronectin, plus a fibroblast-derived extracellular matrix, which we have termed cardiogel. Myocytes cultured on cardiogel were characterized by greater cellular area and volume when compared to cells cultured on the other single-component matrices. Spontaneous contractile activity appeared first in the cells grown on cardiogel, sometimes as early as the first day post-plating, in contrast to day three in the cells cultured on laminin. Measurements of cardiac myocyte contractility i.e. percent shortening and time to peak contraction, were made on each of the first four days in each culture. Myocytes cultured on cardiogel developed maximum shortening more rapidly than the other cultures, and an earlier response to electrical pacing. Histochemical staining for myocyte mitochondrial content, revealed that the cardiogel-supported cells exhibited the earliest development of this organelle and, after four days, the greatest abundance. This reflects both a greater cell size, as well as response to increasing energy demands. Due to the increase in volume and contractile activity exhibited by the cardiogel grown myocytes, we employed calcium binding and uptake experiments to determine the comparative cellular capacities for calcium and as an indicator of sarcoplasmic reticulum development. Also whole cell phosphorylation in the presence of low detergent was assayed, to correlate calcium uptake with phosphorylation, in an attempt to examine possible increases in calcium pump number and other phosphorylatable proteins. In agreement with our physical and contractile data, we found that the cells grown on cardiogel showed a greater calcium uptake over the first four days of culture, and increased phosphorylation. However, calcium binding was not dramatically different comparing the three culture matrices. Based on our data, the fibroblast-derived cardiogel is the matrix of choice supporting earliest maturation of neonatal cardiomyocytes, in terms of spontaneous contractions, calcium handling efficiency, cell size and development of a subcellular organelle, the mitochondrion. PMID- 9865465 TI - L-selectin-mediated lymphocyte aggregation: role of carbohydrates, activation and effects on cellular interactions. AB - L-selectin on lymphocytes reacts with glycosylated ligands on high endothelial venule walls in lymphoid organs. Through this carbohydrate-dependent interaction, rolling and initial attachment of lymphocytes to endothelium is mediated. Here we have studied an earlier described L-selectin-induced homotypic aggregation, to further elucidate the events that occur after engagement of L-selectin. It was found that the interaction of L-selectin with fucoidan, but not with other carbohydrates, or with monoclonal antibodies directed against the carbohydrate recognition domain of L-selectin, resulted in homotypic aggregation among both B- or T lymphocytes. Importantly, this aggregation was shown to be both lymphocyte function-associated antigen-1 (LFA-1) and calcium-independent. Furthermore, for aggregation metabolic energy was required, and signalling via protein tyrosine kinase appeared to be involved. Neither de novo protein synthesis, protein kinase C mediated signalling, Gi-protein mediated signal transduction, nor calcium mobilization were required for aggregation. During aggregation, L-selectin was not shed from the lymphocyte's cell surface. Finally, it was found that the lymphocyte binding capacity to high endothelial venules on cryostat sections was not altered upon triggering these lymphocytes via L-selectin. Interestingly, L selectin-triggered cells showed increased binding to paracortical areas in peripheral lymph nodes. Our data suggest that signals via L-selectin, might lead to altered expression of cell surface molecules, important in interactions other than the first stage of lymphocyte rolling. PMID- 9865466 TI - A common protocadherin tail: multiple protocadherins share the same sequence in their cytoplasmic domains and are expressed in different regions of brain. AB - To study the diversity of protocadherins, a rat brain cDNA library was screened using a cDNA for the cytoplasmic domain of human protocadherin Pcdh2 as a probe. The resultant clones contained three different types. One type corresponds to rat Pcdh2; the other two types are distinct from Pcdh2 but contain the same sequence in their cytoplasmic domains and part of the 3' flanking sequence. To clarify the structure of the proteins defined by the new clones, a putative entire coding sequence corresponding to one of the clones was determined. The overall structure is essentially the same as Pcdh2, indicating that the proteins defined by this clone, and probably by other clones, belong to the protocadherin family. Two PCR experiments and an RNase protection assay showed the existence of the corresponding mRNAs in rat brain preparations. Human and mouse cDNA clones with the same sequence properties were also isolated. Taken together, these results indicate that the clones are not cloning artifacts and that corresponding mRNAs are actually expressed in brains of various species. The results of in situ hybridization showed that the mRNAs corresponding to these clones were expressed in different regions in brain. Since protocadherins encoded by these mRNAs are likely to have different specificity in their interaction and share a common activity at their cytoplasmic domains, these protocadherins may provide a molecular basis, in part, to support the complex cell cell interaction in brain. PMID- 9865467 TI - Ser752 mutation to Pro or Ala in the beta3 integrin subunit differentially affects the kinetics of cell spreading to von Willebrand factor and fibrinogen. AB - The beta3 cytoplasmic domain of the alpha v beta3 integrin is essential for intracellular signals required for cytoskeletal rearrangements. Expression of beta3Ser752Pro mutation in heterologous cells profoundly affects cell spreading and beta3 localization into focal contacts. However, the beta3Ser752Ala substitution mostly restores normal integrin functions, suggesting that the presence of Pro is responsible for the receptor's loss of function. To further assess the role of the Ser752 of the beta3 cytoplasmic domain in the cytoskeletal organization of adherent cells, we developed a computer-assisted method of image analysis allowing the automatic classification of spread cells according to the quantitative analysis of their cell morphology. We compared adhesion and spreading to von Willebrand factor (vWF) or fibrinogen (Fg) of cells expressing beta3 wild type, beta3Ser752Pro or beta3Ser752Ala mutated integrin subunit as a chimeric alpha v beta3 receptor. The beta3Ser752Ala substitution did not impair the general ability of cells to spread, but resulted in a delayed and reduced spreading on both vWF and Fg. Moreover, the beta3Ser752Ala mutation produced modifications of the morphology of spread cells, suggesting a disorganization of their cytoskeleton. Attachment studies showed that the beta3Ser752Ala mutation did not modify the capacity of cells to attach to the substrate, indicating no change in the ligand binding affinity of the alpha v beta3 integrin. Furthermore, we identified a slight defect of beta3Ser752Pro cell attachment to vWF and Fg, beside their impairment of spreading. Taken together, these results suggest a role of Ser752 of the beta3 cytoplasmic domain in the optimal cytoskeletal organization of adherent cells. PMID- 9865468 TI - GlyCAM-1 supports leukocyte rolling in flow: evidence for a greater dynamic stability of L-selectin rolling of lymphocytes than of neutrophils. AB - L-selectin plays a major role in leukocyte traffic through lymph node high endothelial venules (HEV). We have investigated the role of GlyCAM-1, a major L selectin ligand produced by HEV, in mediating leukocyte rolling under in vitro flow conditions. Purified GlyCAM-1 was found to support tethering and rolling in physiological shear flow of both human and murine L-selectin expressing leukocytes at an efficiency comparable to the HEV-derived L-selectin ligands termed peripheral node addressin (PNAd). Major dynamic differences between L selectin rolling of peripheral blood T lymphocytes and neutrophils expressing similar L-selectin level were observed on GlyCAM-1. Lymphocytes established slower and more shear resistant rolling than neutrophils and could roll on GlyCAM 1 at shear stresses lower than the threshold values required for L-selectin mediated neutrophil rolling. Notably, high stability of L-selectin rolling of lymphocytes requires intact cellular energy, although initial lymphocyte tethering to L-selectin ligands is energy-independent. By contrast, L-selectin mediated rolling of neutrophils is insensitive to energy depletion. The distinct dynamic behavior and energy-dependence of L-selectin rolling in different leukocytes suggest that L-selectin adhesiveness in shear flow is regulated in a cell-type specific manner. The greater stability of L-selectin rolling of lymphocytes on surface-adsorbed GlyCAM-1 may contribute to their selective recruitment at peripheral lymph nodes. PMID- 9865469 TI - Hepatic and serum bile acid compositions in patients with biliary atresia: a microanalysis using gas chromatography-mass spectrometry with negative ion chemical ionization detection. AB - Hepatic and serum bile acids in five patients with biliary atresia were preoperatively determined by microanalysis using gas chromatography-mass spectrometry with negative ion chemical ionization detection. The hepatic content of total bile acids was markedly elevated (3079+/-711 nmol/g protein), most of which were primary bile acids. Accumulation of unconjugated bile acids (2.93% to 4.62% of the total) was observed in the liver tissue of these patients, although only trace amounts were detected in their sera. The ratio of glycine-conjugated to taurine-conjugated bile acids was 0.44+/-0.18 in liver tissue and 0.79+/-0.52 in serum and these values were significantly lower than those of controls. This study has shown that the composition of bile acids in serum does not reflect that in liver tissue faithfully. The accumulation of these hydrophobic bile acids may contribute to initiating or exacerbating liver injury in infants with cholestatic liver diseases. PMID- 9865470 TI - Effects of treatment with nilvadipine on cerebral ischemia in rats. AB - The protective effects of a Ca2+ antagonist, nilvadipine, on focal cerebral ischemia were studied in male spontaneously hypertensive rats. The animals received either nilvadipine (3mg x kg(-1) x day(-1)) or a vehicle subcutaneously. Group 1 (n=11) was treated for 7 days, and Group 2 (n=11) for 14 days. The middle cerebral artery was occluded on the 6th (Group 1) or 13th (Group 2) day of the treatment, and neuropathological outcomes were quantified 24 hours later. The mean arterial blood pressure was significantly reduced with nilvadipine to normal levels. The % infarct volumes of Groups 1 (37+/-2) and 2 (34+/-3) were significantly less than those of their controls (39+/-3 [n=11] and 40+/-4 [n=12], respectively), although the difference between Groups 1 and 2 was not significant. When infarct areas were compared in each of 8 coronal sections, the infarct size had decreased in the 5 posterior sections in Group 2, but only in 2 sections of Group 1. A significant decrease in the edema volumes was observed in Group 2, but not in Group 1. Thus, nilvadipine provided protective effects against cerebral ischemia in rats having chronic hypertension, and the effects were dependent on the duration of treatment. PMID- 9865471 TI - Age-related histologic alterations after prednisolone therapy in children with IgA nephropathy. AB - To examine whether the age of onset of clinical symptoms in childhood IgA nephropathy may affect changes of histologic alterations after receiving prednisolone therapy, an evaluation of glomerular lesions seen in biopsy specimen was done. Eighteen children with IgA nephropathy met study criteria. They received alternate-day prednisolone therapy within a month after the first renal biopsy. Renal biopsies were done at presentation and repeated at a mean interval of 23 months. The patients were grouped as follows: Group A, 8 cases which showed clinical symptoms at the age of 9 or under; Group B, 10 cases which showed the symptoms at the age of 10 or over. At the initial presentation, histologic indices including a percentage of mesangial area occupying glomeruli (the M/G ratio) in the 2 groups did not show a significant difference. The activity score and the M/G ratio in the group A decreased significantly at the second biopsies (4.6+/-0.9 vs. 1.8+/-1.0 and 25.7+/-6.1% vs. 21.4+/-2.7%, respectively), while in the group B did not. These observations may indicate the age of onset of clinical symptoms in childhood IgA nephropathy affects changes of histologic alterations after receiving prednisolone therapy. PMID- 9865472 TI - Concentration of metal elements in the blood and urine in the patients with cementless total knee arthroplasty. AB - Titanium (Ti), cobalt (Co) and chromium (Cr) element concentrations in the whole blood and urine specimen in 40 patients with cementless total knee arthroplasty were determined by the electrothermal atomic absorption spectrophotometry. Their ages ranged from 55 to 78 years (mean, 65 years). Twenty of them had loosening of prosthesis and underwent revision surgery, including 4 subjects with Ti-6Al-4V alloy prosthesis and 16 subjects with Co-Cr-Mo alloy prosthesis. The other 20 patients had well-functioning stable prosthesis, including 5 subjects with Ti-6Al 4V alloy prosthesis and 15 subjects with Co-Cr-Mo alloy prosthesis. The mean duration of prostheses implantation in patients with loosened or well-functioning prostheses were 6.5 and 4.0 years, respectively. The control group consisted of 20 age-matched normal subjects who did not undergo any metal implant surgery. Analysis of variance showed that the metal element concentrations in the whole blood, either Co, Cr or Ti, was statistically higher in the patients with loosened prosthesis than the other two groups. However, the metal element concentrations in the urine did not show any difference. The linear regression analysis showed a moderate positive relationship between the metal element concentrations, either Co or Cr elements, in whole blood and urine only in the patients with loosened prostheses. In conclusion, elevated concentration of metal elements may indicate a loosening of prosthesis while the clinical significance of the metal element concentration in the urine needs further investigation. PMID- 9865473 TI - Utilising an ileal segment in the posterior urethral replacement procedures using anal sphincter to provide continence in dogs. AB - We aimed to replace an ileal segment in the place of posterior urethra using the anal sphincter as a continence mechanism. The experiment was carried on three male street dogs. In the first stage, only urethral replacement with an ileal segment was done and pulled through the anal sphincter in a male dog to see if anal sphincter would do any help for continence. A protruding stoma was created on the perineum. Perineal end of the ileal segment was sutured to the bulbous urethra in the other two male dogs to provide urethral patency in the second step of the operation. The dog in which the first operation was made gained continence on the 12th postoperative day. The other two male dogs, in which ileourethral anastomosis were made, became continent on the postoperative 12th and 15th days. No residual urine was found by catheterisation performed after urination. In controls, neither the stoma nor the anastomosis sites developed stenosis. This procedure may be applied in patients with complete incontinence who can not be corrected by any other surgical procedures, and a very good cosmetic result may be obtained. PMID- 9865474 TI - Deconvolution of chemiluminescent emission curve associated with phagocytosis into three logarithmic normal distributions. AB - The chemiluminescent emission reaction dependence on the activity of phagocytosis is well known. However, this method is not used to diagnostically in clinical assessment because the relationship between phagocytizing activity and chemiluminescent intensity has not been clearly established. Therefore, we attempted to analyze quantitatively the chemiluminescent emission curve by the phagocytosis of leukocytes. Mathematical assessment of the emission curve with respect to time was performed by fitting the curve to several regression models using the unweighed non-linear least squares method. A triple logarithmic normal distribution model provided a reasonable goodness of fit to the measured emission curve. The first component, about 5% of the calculated total counts, was assumed to arise from monocytes activity, the second component, about 20% from eosinocytes activity and the third component, up to 75%, from neutrophils activity. This method seems promising as a means for assaying whole blood without the need for pretreatment and for the providing a valid index that is independent of the technical differences between laboratories. PMID- 9865475 TI - A transient emergence of hepatic granulomas in a patient with chronic hepatitis B. AB - So far there were no reports but one on a hepatic granuloma in chronic hepatitis B virus (HBV) infection. We present a case of chronic hepatitis B with a transient emergence of hepatic granulomas. The case was a 35-year-old male who had chronic hepatitis with persistent hepatitis B surface antigen in the sera. A liver biopsy showed noncaseating granulomas in the parenchyma and a mild portal enlargement with mononuclear cell infiltration. The cellular components of the granulomas were mainly cluster of differentiation 68-positive macrophages with a few lymphocytes in the periphery. However, no granulomas were found in a liver specimen obtained three weeks after the first liver biopsy. Possible disorders causing hepatic granulomas such as tuberculosis, sarcoidosis, drugs and other infectious diseases were ruled out by clinical, serological and histopathological examination. Thus it is possible that the transient emergence of hepatic granulomas is a phenomenon related to chronic HBV infection. PMID- 9865477 TI - Evaluation of sex difference in tissue repair following acute carbon tetrachloride toxicity in male and female Sprague-Dawley rats. AB - Cellular regeneration and tissue repair greatly influence the outcome of acute carbon tetrachloride (CCl4) hepatotoxicity. This study examined the temporal kinetics of cellular regeneration and tissue repair processes in male and female Sprague Dawley (SD) rats following an acute CCl4 exposure (0.8 ml/kg, i.p.). In female rats, hepatic damage peaked at 24 h following the treatment and was approximately 2.5-fold (AST 2.7-fold, ALT 2.3 fold) greater than the damage observed in male rats. The hepatic damage in male rats appeared to peak by 3 h post-exposure and did not significantly change through the 36-h time-point. The activity of cytochrome P 4502E1 was 20% greater in male rats and did not correlate with the magnitude of hepatic damage. Morphometric analysis of cell cycle indices revealed that cellular regeneration was significantly greater in female rats as compared to male rats at 48 h and corresponded proportionally to the extent of liver damage. This study demonstrated that female SD rats respond more severely to acute CCl4 hepatotoxicity than male SD rats and the extent of tissue repair and cellular regeneration was greater in female rats. Furthermore, our results suggest that tissue repair is unlikely to result in accounting for the different responses exhibited by male and female SD rats to CCl4 hepatotoxicity. PMID- 9865476 TI - Methapyrilene hepatotoxicity is associated with oxidative stress, mitochondrial disfunction and is prevented by the Ca2+ channel blocker verapamil. AB - Methapyrilene (MP) is an unusual hepatotoxin in that it causes periportal necrosis in rats. The mechanism of acute methapyrilene hepatotoxicity has, therefore, been investigated in cultured male rat hepatocytes. Addition of methapyrilene to rat hepatocytes resulted in a time- and dose-dependent loss in cell viability between 4 and 8 h of incubation as judged by cellular enzyme leakage. The cytochrome P450 (CYP) inhibitor metyrapone protected against methapyrilene-mediated toxicity suggesting that MP is metabolised by CYP for toxicity. The concentration-dependent protection from methapyrilene toxicity afforded by metyrapone correlated with an inhibition of microsomal CYP2C11 associated androstenedione 16alpha hydroxylase activity, and hepatocytes prepared from hypophysectomised rats (containing reduced levels of microsomal immunodetectable CYP2C11 and associated androstenedione 16alpha hydroxylase activity) showed resistance to the toxic effects of methapyrilene. These data suggest that the toxicity of methapyrilene is predominantly dependent on the CYP2C11 isoform. Treatment of hepatocytes with a toxic concentration of MP caused oxidative stress as indicated by increases in NADP+ levels within 2 h and cellular thiol oxidation as evidenced by a reduction--but not complete loss--in glutathione levels. Methapyrilene hepatotoxicity was associated with an early loss in mitochondrial function, as indicated by mitochondrial swelling and significant losses in cellular ATP within 2 h. Co-incubation of methapyrilene treated hepatocytes with inhibitors of inner mitochondrial transition permeability pore opening--cyclosporin A or the thiol reductant dithiothreitol- abrogated cell death suggesting that pore opening and loss of mitochondrial Ca2+ homeostasis play a significant role in methapyrilene-mediated cell death. Co incubation of methapyrilene-treated hepatocytes with the phenylalkylamine calcium channel blocker verapamil--but not by treating cells in a nominally calcium-free medium--also abrogated cell death, suggesting that if Ca2+ is involved in cell killing then it is dependent on an intracellular Ca2+ pool. Pre-treatment of hepatocytes for 1 h with verapamil--to inhibit intracellular Ca2+ pool filling- increased the potency of verapamil protection against methapyrilene toxicity by approximately 100-fold. Taken together, these data indicate that methapyrilene intoxication leads to mitochondrial disfunction and suggest a critical role for a loss of mitochondrial Ca2+ homeostasis in this model of hepatocyte death. PMID- 9865478 TI - Penetration of cisplatin into mouse brain by lipopolysaccharide. AB - We investigated the penetration of cisplatin into the mouse cerebral cortex-rich region (CCR) induced by lipopolysaccharide (LPS). With the injection of cisplatin into mice 3 h after the LPS treatment, platinum was detected in the CCR during the 7 days after the injection, while platinum was not detected in the CCR of cisplatin-injected mice without LPS pretreatment and of mice simultaneous treated with cisplatin and LPS. The N(G)-nitro-L-arginine methyl ester dose-dependently lowered the platinum level. A dose of 5 mg/kg of aminoguanidine reduced the increase in the platinum level of the LPS-treated mouse, and platinum was no longer detected at doses of 20 mg/kg in the aminoguanidine-injected group. At doses of 500 mg/kg aminoguanidine, however, no effect was seen on the platinum level of the CCR induced by LPS. Regarding indomethacin, the injection of 5 mg/kg resulted in a decrease in the platinum content of the CCR, but not undetectable level. These results suggest that LPS increases the penetration of cisplatin into the mouse brain, and platinum may be accumulated in the CCR. Nitric oxide and prostaglandins contribute to the penetration of platinum into the cerebral cortex. PMID- 9865480 TI - In vitro genotoxic effects of the insecticide deltamethrin in human peripheral blood leukocytes: DNA damage ('comet' assay) in relation to the induction of sister-chromatid exchanges and micronuclei. AB - Deltamethrin, a synthetic dibromo-pyrethroid insecticide, is extensively used in agriculture, forestry and in household products because of its high activity against a broad spectrum of insect pests (both adults and larvae), its low animal toxicity and its lack of persistence in the environment. Data on the genotoxicity and carcinogenicity of deltamethrin are rather controversial, depending on the genetic system or the assay used. The aim of this study was to further evaluate the potential genotoxic activity of deltamethrin. The in vitro genotoxicity of deltamethrin has been evaluated by assessing the ability of the insecticide to damage DNA (as evaluated using the single-cell microgel-electrophoresis or 'comet' assay) or induce sister-chromatid exchanges (SCE) and micronuclei (MN) in human peripheral blood leukocytes. All treatments were conducted with and without the presence of an external bioactivation source (+/- S9mix). The results indicate that deltamethrin, in the presence of metabolic activation (+ S9mix), is able to induce DNA damage (double- and single-strand breaks, alkali-labile sites and open excision repair sites) as revealed by the increasing tail moment values observed with increasing doses. The frequency of SCE and MN were not statistically increased in deltamethrin-treated cells as compared to controls, both with and without S9mix. However, lower deltamethrin doses were tested, as compared to 'comet' assay, because of cytotoxicity. PMID- 9865479 TI - Selective induction of apoptosis in mouse and human lung epithelial cell lines by the tert-butyl hydroxylated metabolite of butylated hydroxytoluene: a proposed role in tumor promotion. AB - Butylated hydroxytoluene (BHT) causes lung injury in mice and promotes tumor formation. Hydroxylation of a tert-butyl group on BHT to yield the metabolite, 6 tert-butyl-2-[2'-(2'-hydroxymethyl)-propyl]-4-methylphenol (BHTOH), may be required. BHTOH is more potent than BHT on an equimolar basis in causing lung damage, enhancing lung tumor development, killing isolated bronchiolar non ciliated Clara cells, and inhibiting lung epithelial gap junctional intercellular communication. One mechanism proposed for tumor promoting agents is selective cytotoxicity; killing normal cells allows uninhibited clonal expansion of neighboring initiated cells. We compared the abilities of BHT, BHTOH, and other BHT metabolites to kill non-tumorigenic and tumorigenic mouse and human lung cell lines, and examined the contribution of apoptosis to this cytotoxicity. These cells lack the cytochrome P450 2B isozyme necessary for converting BHT to BHTOH. BHTOH and 4-hydroperoxy-4-methyl-2,6-di-tert-butyl-2,5-cyclohex-adienone+ ++ (BHTOOH) were most toxic, BHT and 2,6-di-tert-butyl-1,4-benzoquinone (BHTQu) were less potent, and 4-methyl BHT metabolites that are not pneumotoxic were ineffective. BHTOH most strongly induced apoptosis, based on nuclear condensation and transmission electron microscopy. Non-tumorigenic cells were as susceptible to cell death as the neoplastic cell lines when apoptosis and necrosis are not distinguished, but more sensitive to BHTOH-induced apoptosis. An apoptotic mechanism may underlie the lung tumor promoting actions of BHTOH. PMID- 9865481 TI - Effects of dichloroacetate on glycogen metabolism in B6C3F1 mice. AB - Dichloroacetate (DCA) is a by-product of drinking water chlorination. Administration of DCA in drinking water results in accumulation of glycogen in the liver of B6C3F1 mice. To investigate the processes affecting liver glycogen accumulation, male B6C3F1 mice were administered DCA in drinking water at levels varying from 0.1 to 3 g/l for up to 8 weeks. Liver glycogen synthase (GS) and glycogen phosphorylase (GP) activities, liver glycogen content, serum glucose and insulin levels were analyzed. To determine whether effects were primary or attributable to increased glycogen synthesis, some mice were fasted and administered a glucose challenge (20 min before sacrifice). DCA treatments in drinking water caused glycogen accumulation in a dose-dependent manner. The DCA treatment in drinking water suppressed the activity ratio of GS measured in mice sacrificed at 9:00 AM, but not at 3:00 AM. However, net glycogen synthesis after glucose challenge was increased with DCA treatments for 1-2 weeks duration, but the effect was no longer observed at 8 weeks. Degradation of glycogen by fasting decreased progressively as the treatment period was increased, and no longer occurred at 8 weeks. A shift of the liver glycogen-iodine spectrum from DCA treated mice was observed relative to that of control mice, suggesting a change in the physical form of glycogen. These data suggest that DCA-induced glycogen accumulation at high doses is related to decreases in the degradation rate. When DCA was administered by single intraperitoneal (i.p.) injection to naive mice at doses of 2-200 mg/kg at the time of glucose challenge, a biphasic response was observed. Doses of 10-25 mg/kg increased both plasma glucose and insulin concentrations. In contrast, very high i.p. doses of DCA (> 75 mg/kg) produced progressive decreases in serum glucose and glycogen deposition in the liver. Since the blood levels of DCA produced by these higher i.p. doses were significantly higher than observed with drinking water treatment, we conclude that apparent differences with data of previous investigations is related to substantial differences in systemic dose and/or dose-time relations. PMID- 9865482 TI - Nicotine enantiomers and oxidative stress. AB - Nicotine affects a variety of cellular processes ranging from induction of gene expression to secretion of hormones and modulation of enzymatic activities. The objective of this study was to characterize the toxicity of nicotine enantiomers as well as their ability to induce oxidative stress in an in vitro model using Chinese hamster ovary (CHO) cells. Colony formation assay has demonstrated that ( )-nicotine is the more toxic of the enantiomers. At 6 mM concentrations, (-) nicotine was found to be approximately 28- and 19-fold more potent than (+)-, and (+/-)-nicotine (racemic), respectively. Results also indicated that the toxicity of (+/-)-nicotine is higher than that of (+)-nicotine. (-)-Nicotine at a 10 mM concentration substantially decreased glutathione (GSH) levels (46% decrease). In addition, a 3-fold increase in malondialdehyde (MDA) level was evident in cells after exposure to 10 mM (-)-nicotine. Increased lactate dehydrogenase (LDH) activities in the media demonstrated that cellular membrane integrity was disturbed in nicotine treated cells. In the presence of superoxide dismutase (SOD) and catalase (CAT), the LDH activities returned to control value in 24 h with all concentrations of (-)-, (+)-, and (+/-)-nicotine. The decreases in LDH activities in the presence of the radical scavenging enzymes SOD and CAT suggest that membrane damage may be due to free radical generation. PMID- 9865483 TI - Effects of N-acetylcysteine and 2,3-dimercaptosuccinic acid on lead induced oxidative stress in rat lenses. AB - Lead (Pb) is known to disrupt the pro-oxidant/anti-oxidant balance of tissues which leads to biochemical and physiological dysfunction. The present study investigated the effects of exposure on the redox status of the lenses of Fisher 344 rats and examined whether antioxidant or chelator administration reversed these changes. Animals were given 5 weeks of 2000 ppm Pb exposure followed by 1 week of either antioxidant, chelator or distilled water administration. Glutathione (GSH) and cysteine (CYS) levels decreased in the Pb-exposed group. N acetylcysteine or 2,3-dimercaptopsuccinic acid (Succimer) supplementation following Pb intoxication resulted in increases in the GSH and CYS levels. Protein bound glutathione (PSSG) and cysteine (PSSC) increased following Pb exposure. In the Succimer-treated animals, the PSSG decreased significantly. The glutathione disulfide (GSSG) levels remained unchanged. Malondialdehyde (MDA) levels, a major lipid peroxidation byproduct, increased following Pb exposure and decreased following Succimer treatment. Our results suggest that antioxidant supplementation, as well as chelation, following Pb exposure may enhance the reductive status of lenses. PMID- 9865484 TI - Comparative aluminum mobilizing actions of deferoxamine and four 3-hydroxypyrid-4 ones in aluminum-loaded rats. AB - The efficacy of the Al chelating drugs deferoxamine (DFO) and the hydoxypyridones (HPs): 1,2-dimethyl-3-hydroxypyrid-4-one (L1), 1-[3-hydroxy-2-methyl-4 oxopyridyl]-2-ethanesulfonic acid (L6), 1-benzyl-(4-carboxylic acid)-3-hydroxy-2 methyl-4-oxopyridine (Bzcal) and 1-(p-methylbenzyl)-2-ethyl-3-hydroxypyrid-4-one (MeBzEM) in increasing Al excretion and reducing tissue Al accumulation has been compared in adult male rats which had previously received Al nitrate nonahydrate i.p. at 0.16 mmol/kg per day for 2 months. At the end of this period, DFO was injected s.c. and the HPs were given by gavage at 0.89 mmol/kg per day for five consecutive days. Total urines were collected 24 h after each chelator administration. Following chelation treatment animals were killed and samples of brain, bone, liver, kidney, and spleen were collected. DFO administration increased to about 4 x the cumulative urinary Al elimination for 5 days, while the excretion of Al into urine caused by Bzcal, L1, and MeBzEM administration was about twice that of the control group. On the other hand, treatment with Bzcal, DFO, and MeBzEM for 5 days significantly reduced the Al levels in bone by 31, 33, and 29%, and the Al concentrations in brain by 46, 69, and 71%, respectively. These results suggest that oral administrations of MeBzEM and Bzcal can be potential alternatives to parenteral administration of DFO in Al removal. PMID- 9865485 TI - 2-Nitropropane-induced lipid peroxidation: antitoxic effects of melatonin. AB - The degree of lipid peroxidation (LPO) as indicated by the levels of thiobarbituric acid reactive substances, malondialdehyde (MDA) and 4 hydroxyalkenals (4-HDA), and the activity of sorbitol dehydrogenase (SDH) in serum as parameters of hepatotoxicity were studied in rats treated with a single intraperitoneal (i.p.) injection of the hepatocarcinogen 2-nitropropane (2-NP). Since melatonin, the main secretory product of the pineal gland, has been shown to protect against a number of toxic agents, it was given 30 min before 2-NP to test its protective effect against 2-NP toxicity. Significant increases in LPO in liver (P<0.0001), lung (P<0.05) and kidney (P<0.0001) were observed 24 h after 4 mmol/kg 2-NP while serum SDH activity was increased 470-fold. All parameters showed time (0, 4, 8, 24 h) and dose (0, 1, 2, 3, 4 mmol/kg) dependency. The induction of LPO by 2-NP was significantly reduced in lung and kidney when melatonin (2.5, 5 or 10 mg/kg) was given prior to 2-NP administration. The elevation in serum SDH caused by 2-NP was also reduced when melatonin was given. These findings show that 2-NP induces LPO and that pharmacological levels of melatonin can reduce the toxicity of this hepatocarcinogen. PMID- 9865486 TI - The role of interferon regulatory factors in the interferon system and cell growth control. AB - Complex cellular responses are often coordinated by a genetic regulatory network in which a given transcription factor controls the expression of a diverse set of target genes. Interferon regulatory factor (IRF)-1 and IRF-2 have originally been identified as a transcriptional activator and repressor, respectively, of the interferon-beta (IFN-beta) as well as of IFN-inducible genes. However, these factors have since been shown to modulate not only the cellular response to IFNs, but also cell growth, susceptibility to transformation by oncogenes, induction of apoptosis, and development of the T cell immune response. Furthermore, the evidence suggests that deletion and/or inactivation of the IRF-1 gene may be a critical step in the development of some human hematopoietic neoplasms. Subsequently, these factors have been shown to constitute a family of transcription factors, termed the IRF-family. Recent studies indicate that other IRF family members also involve the regulation of the IFN system and cell transformation. The IRF-family may be examples of transcription factors which can selectively modulate several sets of genes depending on the cell type and/or nature of the cellular stimuli, so as to evoke host defense mechanisms against infection and oncogenesis. PMID- 9865487 TI - Role of the interferon regulatory factors (IRFs) in virus-mediated signaling and regulation of cell growth. AB - As a response to viral infection, cells express the early inflammatory genes that encode small proteins generally called cytokines or chemokines. These protein can activate immune responses to viral infection as well as to modulate directly the outcome of viral infection. The group of proteins with the direct antiviral effects have been called interferons. The stimulation of interferon synthesis in infected cells is regulated on a transcriptional level and two families of cellular transcriptional factors seem to play a critical role in the transcriptional activation of interferon genes. The first one are the proteins of NF-kappaB family and the second is the family of the interferon responsive factors. While both of the types of the transcriptional factors are important for the induction of interferon beta gene, the NF-kappaB factor do not seems to participate in the induction of interferon alpha genes. The present review is focused on the recently identified new members of cellular IRF family and their role in virus mediated response, responses and cell growth. In addition the HHV-8 encoded vIRFs are described. PMID- 9865488 TI - In vivo formation of IRF-1 homodimers. AB - Interferon regulatory factor 1 (IRF-1) is a transcriptional activator which exerts different biological activities. IRF-1 activates interferon induced genes as well as genes which are not directly linked to the interferon system, such as the ICE protease gene. IRF-1 activity is post-transcriptionally regulated in addition to transcriptional regulation by interferons, cytokines, hormones and many other factors. This includes heterodimerisation with activators and repressors of transcription. These protein interactions modulate the transactivating capacity of IRF-1. By using a two-hybrid system, we demonstrate that IRF-1 forms homodimers in vivo. The homodimerization domain was determined to be located in the N-terminal part of IRF-1 which belongs to the DNA-binding domain. Since this sequence is highly conserved between members of the IRF family, our observation raises the question of homodimerization of other IRFs through this domain. PMID- 9865489 TI - Non-B-DNA structures on the interferon-beta promoter? AB - The high mobility group (HMG) I protein intervenes as an essential factor during the virus induced expression of the interferon-beta (IFN-beta) gene. It is a non histone chromatine associated protein that has the dual capacity of binding to a non-B-DNA structure such as cruciform-DNA as well as to AT rich B-DNA sequences. In this work we compare the binding affinity of HMGI for a synthetic cruciform DNA to its binding affinity for the HMGI-binding-site present in the positive regulatory domain II (PRDII) of the IFN-beta promoter. Using gel retardation experiments, we show that HMGI protein binds with at least ten times more affinity to the synthetic cruciform-DNA structure than to the PRDII B-DNA sequence. DNA hairpin sequences are present in both the human and the murine PRDII-DNAs. We discuss in this work the presence of, yet putative, non-B-DNA structures in the IFN-beta promoter. PMID- 9865490 TI - Type I interferon gene expression: differential expression of IFN-A genes induced by viruses and double-stranded RNA. AB - The family of interferon regulatory transcription factors (IRF) participates in the virus-induced and dsRNA-stimulated transcriptional regulation of either type I IFN genes or a definite set of genes which can also be activated by IFN. In this review, we place emphasis on the role of IRF-3 that associates with the coactivators CBP and/or p300, together or not with IRF-7. These complexes bind to the PRDI, PRDI-like domains or to a number of ISRE sequences located in the promoter of these virus-inducible genes. We also discuss the involvement of the IRF-3-related complexes in the differential regulation of IFN-A genes. PMID- 9865491 TI - Transcriptional repression of type I IFN genes. AB - Transcriptional regulation is a consequence of the combination of both activation and repression for establishing specific patterns of eukaryotic gene expression. The regulation of the expression of type I interferon (IFN-A and IFN-B) multigene family is controlled primarily at the transcriptional level and has been widely studied as a model for understanding the mechanisms of stable repression, transient virus induction and postinduction repression of the genes. The positive and negative regulatory elements required for this on/off switch have been defined within a complex 5' upstream region of their transcription start site. The differential expression pattern of type I IFN genes is thought to involve both substitutions in the virus responsive element (VRE) and presence or absence of negatively acting sequences surrounding the VRE. In this review we discuss several mechanisms of negative regulation due to the existence of common or specific elements in the IFN-B and IFN-A genes and we summarize recent studies on transcriptional repressors that bind to these promoters. PMID- 9865492 TI - Cooperative binding of Stat1-2 heterodimers and ISGF3 to tandem DNA elements. AB - Interferon (IFN)-alpha-activated Stat1 homodimers and Stat1-2 heterodimers bind to GAS elements, whereas the transcription factor ISGF3, which contains Stat1, Stat2 and p48, binds to ISREs. We now find that Stat1-2 dimers can form heterotetramers on tandem GAS sites and that the heterotetramers have a much higher binding affinity for a double GAS site than do heterodimers for a single site, suggesting cooperativity mediated through protein-protein interactions. Stat1-2 heterotetramers can also be detected with a single GAS site, again indicating cooperativity mediated through protein-protein interactions. Deleting 40 amino acid residues from the N-terminus of Stat1 abolished Stat1-Stat2 heterotetramer formation, but did not affect heterodimer formation and an N terminal peptide containing the first 120 residues of Stat2 inhibited heterotetramer formation but did not affect heterodimer formation. Thus, the N terminal regions of both Stat1 and Stat2 are important for cooperative DNA binding, and heterodimers probably interact with each other through these regions. Cooperative binding of ISGF3 was also observed using the tandem ISREs from the IFN-alpha responsive promoter of the 6-16 gene. A more abundant and larger complex was formed with a probe containing two ISREs than with a probe containing a single ISRE. The N-terminal regions of both Stat1 and Stat2 are important for the cooperative binding of ISGF3 to tandem ISREs but not to a single site. The cooperative DNA-binding activities of ISGF3 and Stat1-2 dimers are likely to contribute to the transcriptional activation of those IFN-alpha responsive genes that have tandem DNA elements. PMID- 9865493 TI - Selective mRNA degradation by antisense oligonucleotide-2,5A chimeras: involvement of RNase H and RNase L. AB - Antisense oligonucleotides (ON) allow the specific control of gene expression and phosphorothioate derivatives are currently being evaluated for possible clinical applications. Numerous second generation ON analogues with improved pharmacological properties have been described. Most of them, however, do not recruit RNase H, which is known to increase ON potency by eliciting the specific degradation of the target RNA. Silverman, Torrence and colleagues have conjugated 2,5A to natural antisense ON and demonstrated the preferential cleavage of a target RNA in cell-free and intact cell experiments. We have established for the first time that RNase H-incompetent ON, viz. alpha-anomeric ON analogues, can be converted into sequence-specific nucleases upon conjugation to 2,5A. The use of alpha-ON- and beta-ON-2,5A chimeras has allowed us to delineate the part played by RNase H and RNase L in target RNA degradation and translation arrest. Finally, the present studies have revealed limitations which are encountered in the choice of a suitable target for such ON-2,5A chimeras. PMID- 9865494 TI - The Ifi 200 genes: an emerging family of IFN-inducible genes. AB - The biological activities of interferons (IFNs) are mediated by IFN-induced proteins. One family is encoded by several structurally related genes located on murine chromosome 1 (Ifi 200 cluster) and three homologous genes (MNDA, IFI 16 and AIM2) located on human chromosome 1 as well, within a linkage group highly conserved between mouse and human. All the proteins of this family contain at least one copy of a conserved 200 amino acid domain, in addition to other regions that are different or missing among the various family members. Conservation of the 200 amino acid segment, therefore, may be responsible for a common function, while individually expressed domains may afford other tissue- or cell-specific functions. The data available demonstrate that at least two members of the Ifi 200 protein family, p202 and p204, inhibit cell proliferation in vitro. Moreover, high constitutive levels of p204 expression impair normal embryo development in transgenic animals. Here, we will review the principal features of murine and human proteins belonging to this family and their function in the cell growth regulatory activities mediated by IFNs. PMID- 9865495 TI - Interferon-gamma is a target for binding and folding by both Escherichia coli chaperone model systems GroEL/GroES and DnaK/DnaJ/GrpE. AB - IFN-gamma can be physicochemically distinguished from interferons-alpha, -beta or -omega through the loss of its tertiary structure and biological activity upon exposure to acid or heat. This loss is due to the irreversible aggregation of an unfolded or partially folded state. The conformational instability of IFN-gamma is reflected by its impairment to fold properly when overexpressed in Escherichia coli, resulting in its accumulation in cytoplasmic inclusion bodies. Chaperones were originally identified as a heterogeneous group of proteins that mediate the folding and correct assembly of various polypeptide substrates, and protect thermolabile proteins against inactivation. In either of both cases, chaperones prevent irreversible misfolding by assisting the substrate protein along its pathway to a stable tertiary conformation. Among the best characterized chaperones are the Escherichia coli Hsp60 and Hsp70 heat shock protein complexes, i.e., GroEL/GroES and DnaK/DnaJ/GrpE. They exhibit entirely different reaction mechanisms, which, however, both depend on hydrolysis of ATP. The unfolding of recombinant IFN-gamma by acid or heat can be used as a tool to assess its in vitro interaction with each of both chaperone systems at physiological temperature (35 degrees C). Using such an experimental set-up, both the DnaK and GroEL chaperone systems appeared to form complexes with IFN-gamma from which correctly folded protein was released in an ATP-dependent manner. In addition to the biotechnological implication of these observations, the relevance to de novo folding of IFN-gamma is discussed. PMID- 9865496 TI - Production of nitric oxide (NO) in human hydatidosis: relationship between nitrite production and interferon-gamma levels. AB - Human hydatidosis is characterized by a prolonged coexistence of parasite (Echinococcus granulosus) and host without effective rejection. The basis of the immune response of the patient is poorly understood. Previously, we reported the presence of IFN, TNF-alpha and IL-6 activities in the serum of patients with liver and lung hydatidosis. In the present work, we have investigated the production of nitrite (NO2-) in the serum of hydatidic patients carrying hepatic and pulmonary cysts (range 36-300 microM). Our present data show a correlation between the production of nitrite + nitrate (NO2- + NO3-) and that of circulating cytokines IFN and IL-6. In relapsing patients who did not produce IFN and IL-6, the observed serum NO2- concentrations were low (range 10-37.2 microM), as compared to those detected in patients before surgery. Induction of NO synthase in leukocytes from hydatidic patients was induced by stimulating these cells with a specific parasitic antigen, Antigen-5, as assessed by the increased levels of NO3- + NO2- in the range of 60-85 microM for patients with liver hydatidosis, as compared to the 20-25 microM detected in healthy controls. Collectively, our data indicate that NO2- + NO3- levels correlate with IFN levels and immunoreactivity, and overall suggest that IFN-gamma and nitric oxide production together play a role in the host defense mechanisms in human hydatidosis. PMID- 9865497 TI - Effects of IFN alpha on late stages of HIV-1 replication cycle. AB - IFN alpha causes a modest reduction of HIV-1 expression in chronically infected monocytoid U937 cells. However, the ratio between cell-associated and shed viral p24 antigen is altered, being the cell-associated fraction dose-dependently enhanced by IFN. Furthermore, a significant decrease of infectivity of both cell associated and shed material is observed. Transmission electron microscopy of IFN treated cells revealed virus assembly being strongly inhibited, with the production of morphologically altered (tear-drop shaped) virus particles. Proteolytic processing of gag proteins appeared to be normal in IFN-treated cultures. However, virions shed from IFN-treated cells showed a markedly reduced incorporation of virus-specific gp120 and cell-derived ICAM-1 by the virus envelope. Additionally, these particles showed a significantly decreased ability to become bound to CD4+ target cells, accounting for, at least in part, the observed decrease of infectivity. Taken together, the data suggest that, in chronically infected cells, IFN alpha can affect late stages of HIV-1 replication, by inhibiting virus assembly and release, and by reducing the infectivity of shed virions. The latter effect seems to be due, at least in part, to altered incorporation of surface glycoproteins and defective particle formation. The relationship between impaired gp120 incorporation and altered morphogenesis of HIV-1 virions is under investigation. PMID- 9865498 TI - IFN-tau: a novel subtype I IFN1. Structural characteristics, non-ubiquitous expression, structure-function relationships, a pregnancy hormonal embryonic signal and cross-species therapeutic potentialities. AB - IFN-tau (IFN-tau) constitutes a new class of type I IFN which is not virus inducible, unlike IFN-alpha and IFN-beta, but is constitutively produced by the trophectoderm of the ruminant conceptus during a very short period in early pregnancy. It plays a pivotal role in the mechanisms of maternal recognition of pregnancy in ruminants and it displays high antiviral and antiproliferative activities across species with a prominent lack of cytotoxicity at high concentrations in vitro in cell culture and possibly in vivo. It exhibits high antiretroviral activity against HIV and exhibits immunosuppressive activity in a multiple sclerosis model and reduces embryo and fetal mortality by stimulation of IL-10 production. In this review all the biochemical and para-hormonal properties of this novel IFN-tau are described in detail: structural characteristics of proteins and genes, trophoblast expression, regulation of its expression, structure of its gene promoter, its absence in human species and in non-ruminant animals, the evolution of the IFN-tau genes, its structure-function relationships with its three-dimensional structure, structural localization of biological activities, its lack of cytotoxicity and its receptor. Surprisingly, for an IFN, IFN-tau is also a pregnancy-embryonic signal with paracrine antiluteolytic activity. In order to maintain luteal progesterone secretion, IFN-tau inhibits PGF-2alpha pulsatile secretion and oxytocin uterine receptivity in early pregnancy. It is believed to suppress pulsatile release of endometrial PGF-2alpha by preventing oxytocin and estrogen receptor expression. Additionally, it directly regulates prostaglandin metabolism and possibly the PGE:PGF-2alpha ratio. PMID- 9865499 TI - Interferon-delta: the first member of a novel type I interferon family. AB - We have recently described a novel type I interferon (IFN) co-expressed with IFN gamma by the trophectoderm of the pig conceptus between day 12 and day 18 of gestation, a development stage that corresponds to implantation in the uterus. This IFN, now officially named IFN-delta, is recognized as the first member of a novel type I IFN family. This paper reviews the main published data on IFN-delta, together with some new data, showing that IFN-delta, while being a true type I IFN, has some very specific structural and biological properties. Sequences related to IFN-delta coding sequence were found in the genome of man and other ungulates but the only other potentially functional gene was found, so far, in the horse. The pig IFN-delta mature protein, with 149 amino acids, is the smallest of all known type I IFNs. It is unusually rich in cysteines (seven residues), and has a very basic isoelectric point. Recombinant IFN-delta expressed in insect cells is glycosylated and has a high antiviral activity on porcine cells, but not on human cells. It has high antiproliferative activity, which is significantly enhanced in the presence of IFN-gamma. This new IFN was shown to bind on pig cells to the same type I receptor as IFN-alpha. IFN-delta and IFN-gamma genes are co-regulated in the pig trophectoderm, whose cells on day 14-16 of development simultaneously secrete both IFN proteins. The biological role of porcine IFN-delta in early pregnancy has been found unrelated to the known antiluteolytic effect of trophoblastic IFN-tau in ruminants. PMID- 9865500 TI - Anti-inflammatory activity of myricetin-3-O-beta-D-glucuronide and related compounds. AB - OBJECTIVE AND DESIGN: The anti-inflammatory effect of myricetinglucuronide (MGL) was investigated and structurally-related compounds were compared to examine the structure/activity-relationship in carrageenan-induced rat paw edema. MATERIALS AND SUBJECTS: In vitro studies were performed using rat basophilic leukemia (RBL 1) cells, human polymorphonuclear leukocytes (PMNL), COX-1 from ram seminal vesicle, COX-2 from sheep placenta and human venous blood. For the in vivo tests male Wistar rats were used, for the ex vivo test perfused rabbit ears. TREATMENT: 1-300 microg/kg MGL or myricetinmethylglucuronate and 0.1-5 mg/kg other related compounds administered p.o. (carrageenan edema). 5, 50 and 150 microg/kg MGL p.o. for 14 days (Freund's adjuvant arthritis), 5 and 50 microg/kg p.o. for 6 days (ulceration). METHODS: Anti-inflammatory effects were measured in carrageenan edema and in adjuvant arthritis. Incidence of gastric lesions was tested in an ulcerogenicity model in vivo. Influence on COX was determined in the perfused rabbit ear, in PMNL and in a test assay using COX-1 and COX-2. 5-LOX activity was studied using PMNL and RBL-1. The influence on platelet aggregation was evaluated measuring light transmission. RESULTS: MGL exerted a marked and dose-dependent anti-inflammatory effect in acute (carrageenan edema, ED50 15 microg/kg, indomethacin ED50 10 mg/kg) and chronic (adjuvant arthritis, inhibition at 150 microg/kg 18.1 % left paw, 20.6% right paw, indomethacin 3 mg/kg 18.0% and 19.4%)) models of inflammation. In the perfused rabbit ear 1 microg MGL inhibited the release of PGI2, PGD2 and PGE2 to the same extent as 1 microg indomethacin. The inhibition of COX-1 in the intact cell system was IC50 = 0.5 microM, that of indomethacin 0.0038 microM. In the isolated enzyme preparations of COX-1 and COX 2 the IC50 was 10 microM and 8 microM, that of indomethacin 9.2 mM and 2.4 microM. In the RBL-1 and PMNL test assay the inhibition of 5-LOX was 0.1 microM and 2.2 microM. An orally administered dose of 50 microg/kg/day induced no gastric ulcers in rats treated for 6 days. The investigations on carrageenan edema showed a close relationship between the structure of MGL and the anti inflammatory effect. CONCLUSIONS: MGL is a COX-1, COX-2 and 5-LOX inhibitor. In view of the moderate in vitro activity and the very potent in vivo activity an additive mechanism must be involved. Small changes in the molecular structure lead to the loss or reduction of the anti-inflammatory activity. PMID- 9865501 TI - Intracellular mechanisms of nitric oxide plus hydrogen peroxide-mediated neutrophil adherence to cultured human endothelial cells. AB - OBJECTIVE AND DESIGN: We investigated signal-transduction in nitric oxide/hydrogen peroxide (NO/H2O2) mediated neutrophil-endothelial adhesion and P selectin mobilization. MATERIALS AND METHODS: Human endothelial monolayers (HUVEC) were exposed to 0.1 mM H2O2 plus an NO donor, 0.5 mM spermine-NONOate, and second message inhibitors and neutrophil adhesion and P-selectin expression measured. RESULTS: Neutrophil adherence induced by NO/H2O2 was blocked by a PKG inhibitor, (KT5823, 0.5 microM), a PKC inhibitor, (Go6976, 10 nM), a calcium chelator, TMB-8 (0.1 mM) and a K+ channel blocker, glibenclamide, (10 microM), but not by a PKA inhibitor, (H-89, 0.1 microM) or a tyrosine kinase inhibitor, (genistein, I microM). P-selectin expression induced by NO/H2O2 was blocked by KT5823 and Go6976, but not by TMB-8 or glibenclamide. CONCLUSIONS: These data demonstrate that NO/ H2O2 promotes neutrophil-endothelial adhesion through PKG, PKC, calcium, and K+ channels, but not PKA or tyrosine kinase. Conversely, P selectin mobilization requires only PKG and PKC. PMID- 9865503 TI - Activation of nuclear factor kappa B in Crohn's disease. AB - OBJECTIVES AND DESIGN: The location and degree of activation of nuclear factor kappa (NFkappaB), a primary transcription factor that plays a regulating role in immune and inflammatory responses, was determined in Crohn's disease using full thickness specimens of bowel collected at surgery. MATERIALS AND METHODS: Resected specimens of inflamed and non-inflamed bowel were collected from thirteen patients with Crohn's disease and non-inflamed bowel from eleven control subjects. Prepared frozen sections were immunostained using a monoclonal antibody to the activated form of the p65 subunit of NFkappaB and the number of positive staining cells counted using a Lennox graticule. RESULTS: The number of cells positive for activated NFkappaB was significantly increased (p = 0.001 ) in all layers of inflamed Crohn's disease bowel, compared to non-inflamed bowel from controls. There was also a significant increase ( p = 0.009) in the number of positive cells, when compared to non-inflamed bowel from control subjects, in the submucosa of non-inflamed areas of Crohn's disease bowel. Cells positive for activated NFkappaB were provisionally identified by morphological criteria as mostly macrophages with some lymphocytes. There was no activation in endothelia. CONCLUSION: NFkappaB is activated within large mononuclear cells in all layers of inflamed areas of the bowel in Crohn's disease and may represent key events in the inflammatory process. Increased activation in the submucosa of non-inflamed Crohn's disease bowel provides further evidence of early immunological activation in macroscopically and microscopically uninvolved areas and an underlying abnormal immune system in Crohn's disease. PMID- 9865502 TI - Generation and subcellular distribution of histamine in human blood monocytes and monocyte subsets. AB - OBJECTIVE AND DESIGN: This study was designed to establish the sites of formation and storage of histamine and histidine decarboxylase (HDC) in human monocytes and two of their subsets. MATERIALS AND METHODS: The experiments were carried out using monocytes from buffy coats of healthy blood donors. Histamine was quantitated by RIA, HDC activity by the formation of histamine. RESULTS: The monocyte subtype RM3/1 contained significantly more histamine than the subset 27E10 (0.041+/-0.025 vs. 0.005+/-0.004 pg/cell, p < 0.05) and also more HDC activity and HDC mRNA. After fractionation of monocyte homogenates in a discontinuous Percoll gradient or by differential centrifugation more than 80% of both, HDC activity and histamine, were recovered from the cytosolic fractions. About 50% of this histamine was found to be bound to proteins. CONCLUSIONS: In monocytes histamine and HDC are colocalized in the cytoplasm indicating a subcellular distribution different from mast cells or basophils. The data also show that histamine is synthesized by the monocytes themselves. PMID- 9865504 TI - Factors affecting pain intensity in a pain model based upon tonic intranasal stimulation in humans. AB - OBJECTIVE: The present publication describes an inflammatory pain model based on an air-stream introduced in the nasal cavity. The aim of the present study was to investigate the influence of flow, humidity and temperature of the air-stream on the intensity of the evoked pain sensations. METHODS: Six healthy volunteers participated in the study. Pain was produced by a stream of air introduced to the nasal cavity at different flow rates (5, 6 and 81/min), humidity (20 and 80% relative humidity) and temperature (16, 24, and 32 degrees C). The evoked pain was quantified by means of visual analogue scales. RESULTS: The air-stream induced a dull and burning painful sensation. The intensity of pain was found to be related to the air-stream's humidity and temperature. Specifically, a dry air stream (20% relative humidity) evoked significantly stronger pain than a humidified air-stream (80% relative humidity). Pain was significantly greater at a temperature of 32 degrees C than at 24 degrees and 16 degrees C, while the temperatures of 16 and 24 degrees C did not differ with respect to pain intensity. In contrast, a tendency towards stronger pain produced by a flow of 81 compared to that with 51 could not be statistically justified within the present study. CONCLUSIONS: The non-invasive pain stimulus was found to be easily applicable and the evoked pain sensation could be modified by variation of humidity and temperature of the air-stream. PMID- 9865505 TI - The behavioral effect of vasopressin in the ventral hippocampus is antagonized by an oxytocin receptor antagonist. AB - [Arg8]vasopressin improved long-term retrieval processes and relearning in a go no go visual discrimination task when bilaterally microinjected at a dose of 25 pg/animal into the ventral hippocampus of mice, 10 min prior to the retention session. We had shown that this enhancing effect is antagonized by pretreatment with equal or lower doses (25 pg or 1 ng) of the vasopressin V1 receptor antagonist, (d(CH2)5Tyr(Me)-vasopressin). The present study was an attempt to determine whether the vasopressin V2 receptor antagonist or oxytocin receptor antagonist is as effective as the vasopressin V1 receptor antagonist to block the behavioral effect of vasopressin in the ventral hippocampus. We tested the effect of 25 pg of [d(CH2)5-D-Ile2,Ile4,Arg8]vasopressin, a vasopressin V2 receptor antagonist, and [d(CH2)5,Tyr(Me)2,Thr4,Tyr-NH9(2)]ornithine vasotocin, an oxytocin receptor antagonist, under the same experimental conditions as those used to test the effect of the vasopressin V1 receptor antagonist. The results showed that the vasopressin V2 receptor antagonist microinjected into the ventral hippocampus did not alter the enhancing effect of vasopressin on retrieval and relearning. In contrast, the oxytocin receptor antagonist blocked the vasopressin enhancing effect on retention processes. We can conclude from the data that both vasopressin V1 receptors and oxytocin receptors seem to be involved in the enhancing effect of vasopressin on memory retention. In contrast, the vasopressin V2 receptors do not seem to be involved in the effect of the peptide. PMID- 9865506 TI - Effect of tachykinin receptor antagonists in experimental neuropathic pain. AB - The intrathecal effect of 0.1 to 10 microg of RP-67,580 (3aR,7aR)-7,7-diphenyl 2[1-imino-2(2-methoxyphenyl)-ethyl]++ +perhydroisoindol-4-one hydrochloride, CP 96,345 (2S,3S)-cis-(2(diphenylmethyl)-N-[(2-methoxyphenyl) methyl]-1 azabicyclo[2.2.2]octan-3-amine), SR-140,333 (S)-(1-?2-[3-(3,4-dichlorophenyl)- 1 (3-isopropoxyphenylacetyl)piperidin-3-yl]ethyl?-4-phenyl-1 -azonia bicyclo[2.2.2.]-octane,chloride), all neurokinin (NK)1-receptor antagonists, SR 48,968 (S)-N-methyl-N[4-(4-acetylamino-4-[phenylpiperidino)-2-(3,4-dichlorophen yl)-butyl]benzamide, a tachykinin NK2 receptor antagonist and SR-142,801 (S)-(N) (1-(3-(1-benzoyl-3-(3,4-dichlorophenyl) piperidin-3-yl)propyl)-4-phenylpiperidin 4-yl)-N-methyl acetamide, a tachykinin NK3 receptor antagonist, and of their respective inactive enantiomers on thresholds of vocalization due to a mechanical stimulus in mononeuropathic (sciatic nerve ligature) and diabetic rats, was examined. The tachykinin NK1 and the NK2 receptor antagonists were antinociceptive in both models, with a higher effect of the former in diabetic rats. The tachykinin NK3 receptor antagonist was weakly effective in diabetic rats only. This indicates a differential involvement of the tachykinins according to the model of neuropathic pain, suggesting a potential role for tachykinin receptor antagonists in the treatment of neuropathic pain. PMID- 9865507 TI - Head and whole-body jerking in guinea pigs are differentially modulated by 5 HT1A, 5-HT1B/1D and 5-HT2A receptor antagonists. AB - The present study examined the role of 5-hydroxytryptamine 5-HT receptor subtypes on 5-hydroxytryptamine- (5-HT-) mediated myoclonus in guinea pigs, evaluating head and whole-body jerking as two distinct behavioural responses. Myoclonus was induced by the 5-HT precursor L-5-hydroxytryptophan (L-5-HTP) and the non selective 5-HT1A/1B/5-HT2 receptor agonist 5-methoxy-N,N-dimethyl-tryptamine (5 MeODMT). The selective 5-HT1A receptor antagonist WAY100635 (N-[2-[4-(2 methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cycloh exanecarboxamide trihydrochloride) inhibited both head and whole-body jerking. The selective 5 HT1B/1D receptor antagonist GR127935 (N-[4-methoxy-3-(4-methyl-1 piperazinyl)phenyl]-2'-methyl-4'-(5-methyl-1 ,2,4-oxadiazol-3-yl)[1,1'-biphenyl] 4-carboxamide hemifumarate) only inhibited whole-body jerking, which resulted in unmasked head jerking. Co-administration of GR127935 and the selective 5-HT2A receptor antagonist MDL100.151 ((+/-)-alpha-(2,3-dimethoxyphenyl)-1-[-2-(4 fluorphenyl)ethyl]-4-+ ++piperidinmethanol) caused a complete inhibition of whole body as well as head jerking. MDL100.151 had only limited effect on myoclonic jerking when given alone. The inhibitory effects of the 5-HT receptor antagonists on either L-5-HTP- or 5-MeODMT-induced myoclonus were found to be very similar. These data confirm a role for the 5-HT1A and 5-HT1B/1D receptors and suggest a role for 5-HT2A receptors in mediating myoclonus in guinea pigs. Moreover, the study shows that by considering head and whole-body jerking as two pharmacologically distinct behavioural responses, subtype specific 5-HT1A, 5 HT1B/1D and 5-HT2A receptor antagonists can be distinguished. PMID- 9865508 TI - Cranial vascular effects of zolmitriptan, a centrally active 5-HT1B/1D receptor partial agonist for the acute treatment of migraine. AB - The anti-migraine drug zolmitriptan is a novel 5-HT1B/1D receptor partial agonist which, unlike sumatriptan, has been shown to cross the intact blood-brain barrier. In this study we examined whether or not the ability to access the cerebro-vascular intima affects the way in which a centrally-active 5-HT1B/1D receptor agonist influences cranial haemodynamics. The effects of zolmitriptan on carotid arterial blood flow distribution were studied in anaesthetised cats using radiolabelled microspheres. Zolmitriptan (10-1000 microg kg(-1) i.v.) selectively reduced arteriovenous-anastomotic (AVA) conductance producing a maximum decrease of 92.5+/-2.3%. The drug also produced a modest reduction in extra-cerebral conductance (23.9+/-6.5% maximum reduction at 30 microg kg(-1), i.v.), but was without effect on cerebral conductance. Using laser doppler flowmetry in anaesthetised cats, zolmitriptan (1-30 microg kg(-1), i.v.) produced dose dependent decreases in ear microvascular conductance (15+/-5 to 60+/-6%) which mirrored decreases in carotid arterial conductance (12+/-11 to 61+/-5%). By contrast, zolmitriptan at doses up to 1000 microg kg(-1) was without effect on cerebral microvascular conductance. Although zolmitriptan crosses the blood-brain barrier and can therefore access the cerebro-vascular intima, this study suggests that this property does not adversely affect cerebrovascular function. PMID- 9865509 TI - Tranilast, an anti-allergic drug, possesses antagonistic potency to angiotensin II. AB - N-(3',4'-dimethoxycinnamoyl) anthranilic acid (tranilast), an effective anti allergic drug, has successfully prevented restenosis in patients who have undergone percutaneous transluminal coronary angioplasty. To elucidate the mechanism of tranilast, we investigated its antagonistic effect to angiotensin II, which plays a pivotal role in the proliferation of vascular smooth muscle cells, using angiotensin II-induced contractions in human gastroepiploic artery and rabbit aorta. The possible antagonistic effects of other anti-allergic agents such as 4-( p-chlorobenzyl)-2-(hexahydro-1-methyl-1H-azepin-4-yl)-1(2H)-phthal azinone hydrochloride (azelastine), 9-methyl-3-( 1H-tetrazol-5-yl)-4H-pyrido[1,2 a]pyramidin-4-one potassium salt (pemirolast) and disodium cromoglycate were also compared. Tranilast dose-dependently inhibited the angiotensin II-induced contractions in human and rabbit arteries (IC50 = 3.6x10(-5) M and pD'2 = 3.69, respectively). Pemirolast showed a weak antagonistic effect to angiotensin II, but the effective concentration cannot be administered in clinical dosage. Tranilast and pemirolast had no effect on the concentration-contractile response curves for KCI and norepinephrine. Azelastine inhibited angiotensin II-, KCl- and norepinephrine-induced contractions non-specifically, while disodium cromoglycate did not affect these contractile responses. Tranilast but not azelastine showed synergistic action with 2-ethoxy-1-[[2'-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl] 1H-benzimi dazole-7-carboxylic acid (CV- 11974) in antagonizing angiotensin II induced contraction and the inhibitory pattern was similar to that of the non peptide angiotensin II AT1 receptor antagonist CV-11974. These findings indicate that only tranilast possesses the unique ability to antagonize angiotensin II in clinical dosage, which may contribute at least in part to prevention of restenosis after percutaneous transluminal coronary angioplasty. PMID- 9865510 TI - Sex differences in extracellular and intracellular calcium-mediated vascular reactivity to vasopressin in rat aorta. AB - In rat thoracic aorta, contractile responses to arginine vasopressin are two-fold higher in females than in males. To determine the roles of extracellular and intracellular Ca2+ in this sexual dimorphism in vascular function, vascular reactivity and Ca2+ channel function were examined in thoracic aortae of male and female rats. In the presence of diltiazem (10 microM), maximal contraction to vasopressin was reduced to a greater extent in male (65+/-2%) than in female aortae (38+/-1%). Maximal contractile responses to KCl and Bay K 8644 were similar in male and female aortae. Sensitivity to KCI was slightly but significantly higher in male than in female aorta; in contrast, sensitivity to Bay K 8644 was nearly three-fold higher in males than in females. Removal of the endothelium enhanced sensitivity to KCl similarly in male and female aortae. In the presence of simvastatin (60 microM; an inhibitor of intracellular Ca2+ release), reactivity to vasopressin was reduced substantially in female (42+/-1%) but unaltered in male aortae. Removal of the endothelium enhanced the inhibitory effect of simvastatin in both female (73+/-2%) and male aortae (41+/-2%). These findings demonstrate that male aortae depend more upon extracellular Ca2+ influx, whereas female aortae depend more upon intracellular Ca2+ release for vasopressin induced contraction. PMID- 9865511 TI - Lysophosphatidylcholine potentiates vascular contractile responses by enhancing vasoconstrictor-induced increase in cytosolic free Ca2+ in rat aorta. AB - We investigated the effects of palmitoyl-L-alpha-lysophosphatidylcholine on the contractile responses of the endothelium-denuded rat aorta to high K+, noradrenaline, UK14,304 (5-bromo-6-[2-imidazolin-2-ylamino]-quinoxaline) (a selective alpha2 adrenoceptor agonist) and phorbol 12-myristate 13-acetate (PMA). Lysophosphatidylcholine at concentrations from 10(-6) M to 10(-4) M did not contract aortic strips. However, lysophosphatidylcholine strongly potentiated the UK14,304-induced contraction. High K+ - and PMA-induced contractions were also potentiated. In contrast, the noradrenaline-induced contraction was only slightly potentiated by 10(-5) M lysophosphatidylcholine. In fura PE-3-loaded aortic strips, lysophosphatidylcholine (10(-5) M) markedly augmented the increase in both cytosolic free Ca2+ ([Ca2+]i) and contractile tension induced by UK14,304, high K+ and PMA. Nicardipine (10(-7) M) and 10(-6) M Ro-31-8220 (?1-[3 (amidinothio)propyl-1H-indoyl-3-yl]-3-(1-methyl-1H-++ +indoyl-3-yl)-maleimide methane sulfate) strongly inhibited the increase in [Ca2+]i and contractile tension induced by UK14,304 and in the presence of these inhibitors, the enhancing effects of lysophosphatidylcholine were attenuated. However, the enhancing effect on high K+ -induced contraction was not affected by Ro-31-8220. These results suggest that lysophosphatidylcholine may cause an augmentation of the increase in [Ca2+]i induced by UK14,304 which response is depend on protein kinase C activation and in this way potentiate contractile responses in the rat aorta. Protein kinase C independent mechanisms may also be involved in the enhancing effect of lysophosphatidylcholine on smooth muscle contraction. PMID- 9865512 TI - Cardiovascular effects of noradrenaline in hypovolemic haemorrhage: role of inducible nitric oxide synthase. AB - Hypovolemia has been associated with the induction of nitric oxide synthase which is believed to result in an over-production of nitric oxide. In the present study, we have examined the effects of noradrenaline following haemorrhage on cardiac output, blood pressure, mean circulatory filling pressure and vascular resistance in anaesthetized rats after pre-treatment with nitric oxide synthase inhibitor, L-N6-(1-iminoethyl)lysine or dexamethasone. Hypovolemic haemorrhage resulted in induction of nitric oxide synthase, as measured in lungs, and both dexamethasone and L-N6-(1-iminoethyl)lysine inhibited the activity of the inducible form of nitric oxide synthase. An infusion of noradrenaline significantly increased cardiac output, blood pressure and mean circulatory filling pressure in animals pre-treated with L-N6-(1-iminoethyl)lysine and dexamethasone when compared with saline pre-treatment. In addition, the administration of noradrenaline significantly reduced venous resistance in animals pre-treated with L-N6-(1-iminoethyl)lysine when compared with saline pre treatment. The results of this investigation indicated that the impact of noradrenaline on cardiac output, blood pressure and mean circulatory filling pressure was greater in hypovolemic rats treated with L-N6-(1-iminoethyl)lysine or dexamethasone. In addition, we found that in the hypovolemic state, the greater increase in cardiac output during the infusion of noradrenaline after inhibition of nitric oxide synthase was predominantly due to reduced resistance to venous return. PMID- 9865514 TI - Effect of gliclazide treatment on insulin secretion and beta-cell mass in non insulin dependent diabetic Goto-Kakisaki rats. AB - The Goto-Kakisaki rat is a genetic non-overweight model of non-insulin-dependent diabetes mellitus. Adult Goto-Kakisaki rats exhibit a mild basal hyperglycaemia (11 mmol/l) with impaired glucose tolerance, elevated basal plasma insulin level, a failure of insulin release in response to glucose together with a 50% depletion of the total pancreatic beta-cell mass and insulin stores. We have examined the effects of long-term (4 weeks) gliclazide treatment on the severity of diabetes in adult male Goto-Kakisaki rats (10-12 weeks of age). Gliclazide was administered orally (10 mg/kg per day). Gliclazide-treated Goto-Kakisaki rats were evaluated against Wistar and untreated Goto-Kakisaki rats. In the gliclazide treated Goto-Kakisaki rats, basal plasma glucose levels declined progressively reaching 8 mmol/l as a mean at the end of treatment, and their basal insulin levels decreased to values similar to those in non-diabetic Wistar rats. Despite their total pancreatic beta-cell remaining unaffected, their pancreatic insulin stores were twice increased, with a similar improvement of the insulin content per individual beta-cell. Furthermore, the glucose-stimulated insulin release as evaluated in vivo during an intravenous glucose tolerance-test was significantly improved (twice increased) in the gliclazide-treated Goto-Kakisaki rats. This was correlated with a modest but significant enhancement of the early phase of insulin release in vitro (isolated perfused pancreas), in response to glucose. However, the overall insulin response in vitro remained clearly defective with no reappearance of the late phase of insulin release. The in vitro response to arginine (which was basically amplified in the Goto-Kakisaki model) or to gliclazide were kept unchanged after the gliclazide treatment. In conclusion, chronic gliclazide does not exert any beta-cytotrophic effect, but improves beta cell function in the adult Goto-Kakisaki rat as far as it lowers basal insulin release, increases beta-cell insulin stores, and increases the glucose-induced insulin release. PMID- 9865513 TI - Ovarian steroids and stress produce changes in peripheral benzodiazepine receptor density. AB - Although past research has described changes in the density of the peripheral benzodiazepine receptor in brain and in peripheral organs in response to stressors and steroid hormone exposure, their combined influence had yet to be determined. This study examined the effect of swim-stress as a function of ovarian hormone administration on the binding of an isoquinoline carboxamide derivative, [3H]PK 11195, in brain and peripheral tissues. In olfactory bulb and adrenal gland, stress increased peripheral benzodiazepine receptor density in ovariectomized rats with and without estradiol and progesterone replacement injection, even when compared with unstressed animals treated with hormones, where estradiol + progesterone decreased peripheral benzodiazepine receptor number in olfactory bulb, but estradiol and estradiol + progesterone increased it in adrenal gland. In frontal cortex, stress decreased peripheral benzodiazepine receptor number, an effect that was reversed by estradiol. In hippocampus estradiol decreased peripheral benzodiazepine receptor density in unstressed animals and estradiol + progesterone decreased peripheral benzodiazepine receptor number in unstressed and stressed animals. In cerebellum, stress, estradiol and estradiol + progesterone alone decreased peripheral benzodiazepine receptor density. In uterus of unstressed controls, estradiol + progesterone increased peripheral benzodiazepine receptor density, and stress produced a further increase in steroid-treated females. Stress did not affect peripheral benzodiazepine receptor density in kidney, except in animals that received estradiol + progesterone injections, where swim-stress produced a significant decrease in peripheral benzodiazepine receptor density. Thus, steroid hormones regulate peripheral benzodiazepine receptor density in endocrine organs and brain, and the hormonal state of the organism modifies the peripheral benzodiazepine receptor response to stress in a tissue- and brain region-specific manner, suggesting that the peripheral benzodiazepine receptor may play a pivotal role in an integrated response to stress. PMID- 9865515 TI - 17beta-estradiol inhibition of ascorbic acid accumulation in human intestinal Caco-2 cells. AB - We investigated the effect of estrogen on the accumulation of ascorbic acid by human intestinal Caco-2 cells. 17beta-estradiol, synthetic estrogen diethylstilbestrol, and partial agonist tamoxifen were found to inhibit ascorbic acid accumulation in a dose-dependent fashion. The inhibitory effect of estrogens can be observed at as short as 5 min of incubation. An additive effect was observed when they were used in combination. Similar to dietary flavonoids, inhibition was also observed in two other intestinal cell lines, HT-29 and IEC-6. These chemicals affected both Na+ -dependent and Na+ -independent(K+ substituting Na+) accumulation of ascorbic acid and did not affect the efflux of accumulated ascorbic acid. Kinetic analysis of diethylstilbestrol showed a non-competitive inhibition with an apparent Ki of 23 microM. The hormone-ascorbic acid interaction in the intestinal cell could help to explain the known reduction in blood ascorbic acid level among oral contraceptive users and female guinea pigs given contraceptive hormones. PMID- 9865516 TI - Agonist and antagonist effects of histamine H3 receptor ligands on 5-HT3 receptor mediated ion currents in NG108-15 cells. AB - The ability of histamine H3 receptor ligands to interact with 5-HT3 receptors in NG108-15 cells was studied using the whole cell patch clamp recording technique. Imetit, a histamine H3 receptor agonist, generated inward currents and exhibited weak partial agonist activity at the 5-HT3 receptor (EC50 = 11.8 microM). Imetit induced currents were slow to desensitize and at a high concentration reduced in size. The histamine H3 receptor antagonists iodophenpropit and thioperamide did not generate inward currents but were able to inhibit 5-hydroxytryptamine (5-HT) responses with an IC50 of 1.57+/-0.3 microM and 13.7+/-3.5 microM, respectively. Thioperamide is probably a non-competitive antagonist which may have more than one binding site on the receptor. PMID- 9865517 TI - Nature of methamphetamine-induced rapid and reversible changes in dopamine transporters. AB - The nature of methamphetamine-induced rapid and transient decreases in dopamine transporter activity was investigated. Regional specificity was demonstrated, since [3H]dopamine uptake was decreased in synaptosomes prepared from the striatum, but not nucleus accumbens, of methamphetamine-treated rats. Differences among effects on dopamine transporter activity and ligand binding were also observed, since a single methamphetamine administration decreased [3H]dopamine uptake without altering [3H]WIN35428 ([3H](-)-2-beta-carbomethoxy-3-beta-(4 fluorophenyl)tropane 1,5-naphthalenedisulfonate) binding in synaptosomes prepared 1 h after injection. Moreover, multiple methamphetamine injections caused a greater decrease in [3H]dopamine uptake than [3H]WIN35428 binding in synaptosomes prepared I h after dosing. Finally, decreases in [3H]dopamine uptake, but not [3H]WIN35428 binding, were partially reversed 24 h after multiple methamphetamine injections. Western blotting indicated that saline- and methamphetamine-affected dopamine transporters co-migrated on sodium dodecyl sulfate (SDS) gels at approximately 80 kDa, and that acute, methamphetamine-induced decreases in [3H]dopamine uptake were not due to loss of dopamine transporter protein. These findings demonstrate heretofore-uncharacterized features of the acute effect of methamphetamine on dopamine transporters. PMID- 9865518 TI - Regulation of noradrenaline release by S-nitroso-cysteine: inhibition in PC12 cells in a cyclic GMP-independent manner. AB - Nitric oxide (NO), including NO free radicals (*NO) and peroxynitrite (OONO-), modulates the release of neurotransmitters from neuronal tissues. Although we reported that S-nitroso-cysteine stimulated noradrenaline release in brain slices, we now show that only S-nitroso-cysteine inhibits noradrenaline release from PC12 cells. S-Nitroso-cysteine inhibited, in a dose-dependent manner (up to 0.6 mM), the Ca2+ -dependent [3H]noradrenaline release induced by ionomycin, adenosine 5'-O-(3-thiotriphosphate), or high K+, from PC12 cells labeled with [3H]noradrenaline. Sodium nitroprusside, S-nitroso-N-acetylpenicillamine, and 1 hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene, which specifically release NO free radicals in neutral buffer, had minimal effects on [3H]noradrenaline release, although they markedly stimulated cyclic GMP accumulation. 3 Morpholinosydonimine, which releases peroxynitrite, had no effect on either [3H]noradrenaline release or cyclic GMP accumulation. S-Nitroso-cysteine inhibited phorbol 12-myristate 13-acetate- and mastoparan (wasp venom toxin) induced [3H]noradrenaline release. These findings suggest that 1) S-nitroso cysteine, but not other NO donors, inhibits some common process occurring during noradrenaline release in PC12 cells, 2) neither NO radicals, peroxynitrite, nor cyclic GMP mediate the inhibitory effects of S-nitroso-cysteine in PC12 cells. PMID- 9865519 TI - Cyclooxygenase inhibition reveals synergistic action of vasoconstrictors on mesangial cell growth. AB - Since endogenous vasoconstrictors promote mesangial cell growth and increase the biosynthesis of antiproliferative prostaglandins, the effects of cyclooxygenase inhibition on mesangial cell proliferation should be strongly dependent on the prevailing levels of neuroendocrine vasoconstrictors. We compared the effects of indomethacin (10(-6) M), a cyclooxygenase inhibitor, on [3H]thymidine incorporation by cultured rat mesangial cells in the presence of various combinations of angiotensin II (10(-10) M), [Arg8]vasopressin (10(-11) M), (-) norepinephrine (10(-8) M) and endothelin-1 (10(-11) M). Indomethacin did not enhance [3H]thymidine incorporation in cells treated with each individual vasoconstrictor, or in cells treated with two-way combinations with the exception of modestly increased [3H]thymidine incorporation in cells treated with angiotensin II + (-)-norepinephrine or [Arg8]vasopressin + (-)-norepinephrine. In contrast, in cells treated with any three-way or the four-way combination, indomethacin markedly increased [3H]thymidine incorporation. Importantly, a highly significant interaction (P<0.0001) was observed for thymidine incorporation between the number of vasoconstrictors present and indomethacin treatment, thus demonstrating that cyclooxygenase inhibition reveals a synergistic action of vasoconstrictors on the DNA synthesis in mesangial cells. PMID- 9865520 TI - Effect of SK&F 96365 on extracellular Ca2+ -dependent O2- production in neutrophil-like HL-60 cells. AB - Store-operated Ca2+ entry is referred to a capacitative current activated by Ca2+ -stores depletion in various non-excitable cells. Neutrophil-like HL-60 cells responded to N-formyl-L-Methionyl-L-Leucyl-L-Phenylalanine (fMLP) by an early O2- production preceded by a [Ca2+]i rise. Cell stimulation in the absence of extracellular Ca2+ resulted in a major reduction of [Ca2+]i rise and O2- production. A purported inhibitor of store-operated Ca2+ entry, SK&F 96365 (1 (beta-(3-(4-methoxy-phenyl)propoxyl)-4-methoxy-phenetyl)- 1H-imidazole hydrochloride), inhibited extracellular Ca2+ -dependent [Ca2+]i rise by 30% but did not alter O2- production. In conclusion, SK&F 96365 did not modify extracellular Ca2+ -dependent O2- production, despite a significant but limited reduction in fMLP-activated membrane Ca2+ fluxes which can be ascribed to store operated Ca2+ entry. Furthermore, Ca2+ influx is necessary for a full induction and maintenance of the biological response. PMID- 9865521 TI - The human 5-ht5A receptor couples to Gi/Go proteins and inhibits adenylate cyclase in HEK 293 cells. AB - The G protein coupling of human 5-hydroxytryptamine5A (h5-ht5A) receptors was investigated in stably transfected human embryonic kidney (HEK) 293 cells, using radioligand and guanosine-5'[gamma-35S]thiotriphosphate binding to membranes and cyclic adenosine monophosphate measurements in cells. 5-Carboxamido[3H]tryptamine bound to high- and low-affinity sites on h5-ht5A-HEK 293 cell membranes. Guanylyl imidodiphosphate addition and pertussis toxin pre-treatment abolished high affinity binding, indicating coupling to G proteins of the Gi/Go family. [N methyl-3H]Lysergic acid diethylamide bound to a single site; guanylyl imidodiphosphate and pertussis toxin did not alter lysergic acid diethylamide affinity. 5-Hydroxytryptamine stimulated guanosine-5'[gamma-35S]thiotriphosphate binding to 130% over basal and this effect was completely abolished by pertussis toxin. Various 5-hydroxytryptamine receptor ligands were tested for inhibition of 5-carboxamido[3H]tryptamine binding and in guanosine-5'[gamma 35S]thiotriphosphate binding assays. 5-Hydroxytryptamine consistently inhibited forskolin-induced cyclic adenosine monophosphate formation by 25% in h5-ht5A-HEK 293 cells; no effect was detected on basal cyclic adenosine monophosphate levels, on intracellular Ca2+ concentration or arachidonic acid release. Our studies demonstrate functional coupling of the h5-ht5A receptor to pertussis toxin sensitive G proteins and to inhibition of adenylate cyclase activity. PMID- 9865522 TI - Novel mechanism by which hemoglobin induces constriction of cerebral arteries. AB - Since oxyhemoglobin (OxyHb) is implicated in the pathogenesis of cerebral vasospasm, we have investigated the role of protein tyrosine phosphorylation in OxyHb-mediated signalling in canine cerebral arteries and cultured canine cerebrovascular smooth muscle cells. OxyHb produced a contraction of basilar artery preparations, which was reversed by genistein, an inhibitor of tyrosine kinases, and PD098059, an inhibitor of mitogen-activated protein kinase. In cerebrovascular smooth muscle cells, OxyHb induced tyrosine phosphorylation of 42, 46, 54-60 and 80-100 kDa proteins with a time-course which paralleled the contractile action of OxyHb, suggesting that these events might be functionally linked. The 42 and 60 kDa proteins were immunologically related to the mitogen activated protein kinase, extracellular signal regulated protein kinase (ERK2), and to p60c-Src (c-Src), respectively. The increase in protein tyrosine phosphorylation was attenuated by genistein, and the phosphorylation of the 42 kDa protein (ERK2) was inhibited by PD098059. These results suggest that OxyHb mediated signalling utilizes a protein tyrosine kinase-based mechanism. PMID- 9865523 TI - Adenosine kinase inhibitors attenuate opiate withdrawal via adenosine receptor activation. AB - Previous studies have demonstrated a role for adenosine in mediating opiate effects. This study examines the effects of indirect activation of adenosine receptors, via treatment with adenosine kinase inhibitors, on the expression of opiate withdrawal in mice. Mice receive chronic morphine treatment via implantation of subcutaneous morphine pellets (75 mg) for 72 h. Mice then receive parenteral treatment with adenosine kinase inhibitors, either 5'-amino-5' deoxyadenosine (2, 5, 20, 40 mg/kg, intraperitoneal or i.p.) or iodotubericidin (1, 2, 5 mg/kg, i.p.), followed by naloxone injection and opiate withdrawal signs are measured over 20 min. Both adenosine kinase inhibitors significantly reduce the following opiate withdrawal signs in a dose-dependent manner compared to vehicle: withdrawal jumps, teeth chattering, forepaw tremors, and forepaw treads. Additionally, 5'-amino-5'-deoxyadenosine significantly reduces withdrawal-induced diarrhea and weight loss. Effects of 5'-amino-5'-deoxyadenosine (40 mg/kg) on opiate withdrawal signs appear to be mediated via adenosine receptor activation as they are reversed by pretreatment by adenosine receptor antagonist caffeine (20 mg, i.p.) but not by selective phosphodiesterase inhibitor Ro 20-1724 (10 mg/kg, i.p.). Adenosine receptor activation via adenosine kinase inhibitor treatment attenuates opiate withdrawal and these agents may be generally useful in the treatment of drug withdrawal syndromes. PMID- 9865524 TI - Modifications of dopamine D1 receptor complex in rats self-administering cocaine. AB - Cocaine is spontaneously and experimentally self-administered and, when given repeatedly, it induces a stable form of sensitization to a previously assessed minimum active dose. In the present study, triads of rats chronically implanted with a jugular catheter were treated as follows: one animal was trained to self inject cocaine, while the other two passively received either cocaine or saline whenever the self-administering rat completed the response requirement. After 30 days of stable responding, the animals were sacrificed and dopamine D1 receptor density and adenylyl cyclase activity were measured in different brain areas. Animals receiving cocaine (both self-administering and yoked) showed a down regulation of dopamine D1 receptor number and of dopamine stimulated adenylyl cyclase activity in the nucleus accumbens, as compared to saline rats. In the olfactory tubercle, dopamine stimulated adenylyl cyclase activity appeared selectively and significantly down-regulated in self-administering animals. PMID- 9865525 TI - Interaction between adenosine A1 and A2 receptor-mediated responses in the rat hippocampus in vitro. AB - Previous work has been carried out on the effects of adenosine on transmitter release and on the excitability of postsynaptic neurones, but little is known about the effects of adenosine on the coupling between the two. In this study, we examine the effects of specific adenosine receptor agonists and antagonists on the population excitatory postsynaptic potential (population EPSP) slope, the population spike amplitude, and the relationship between the two (E-S coupling) in the CA1 area of rat hippocampus. Activation of adenosine A1 receptors by adenosine or the selective agonist N6-cyclopentyladenosine resulted in a decrease of the population spike amplitude by a greater extent than could be accounted for by the decrease in population EPSP slope, resulting in a dissociation in the E-S relationship, reflected as a right-shift in the E-S curve. Activation of adenosine A2A receptors by the selective agonist 2-p-(2 carboxyethy)phenethylamino-5'-N-ethylcarboxamidoadeno sine (CGS 21680), or blockade by antagonists ZM 241385 and CP 66713 had no effect on evoked responses. However, when both adenosine A1 and A2A receptors were activated at the same time, a significant attenuation of the inhibitory effects of N6 cyclopentyladenosine on population spike amplitude was observed, resulting in a left-shift in the E-S curve. Intracellular recording indicated that N6 cyclopentyladenosine raised the threshold for spike induction by pulses of depolarising current, even at a concentration which did not produce hyperpolarisation of the neurone. At 30 nM, CGS 21680 prevented this effect of N6 cyclopentyladenosine, and this apparent antagonism was prevented by the A2A receptor antagonist ZM 241385. The results show that adenosine A1 receptors change the coupling between presynaptic transmitter release and postsynaptic cell firing, and that this effect is attenuated by A2A receptor activation. PMID- 9865526 TI - Morpholin-2-yl-phosphinic acids are potent GABA(B) receptor antagonists in rat brain. AB - The pharmacological properties of morpholin-2-yl-phosphinic acids were evaluated on GABA(B) receptors. In rat neocortical slices maintained in Mg2+-free Krebs medium, baclofen, a GABA(B) receptor agonist, produced a concentration-dependent depression of the frequency of spontaneous discharges with an EC50 of 14 +/- 5.5 microM, which was antagonised reversibly by the morpholin-2-yl-phosphinic derivatives. The order of potency was 3-[(3S,6R)-6 [(cyclohexylmethyl)hydroxyphosphinoylmethyl- morpholin-3-yl]benzoic acid (CGP 76290A) (pA2 = 7.1 +/- 0.05) > its enantiomer 3-[(3R,6S)-6 [(cyclohexylmethyl)hydroxyphosphinoylmethyl]-++ +morpholin-3-yl]benzoic acid (CGP 76291A) (pA2 = 6.8 +/- 0.1) > cyclohexylmethyl-[(2R',5S')-5-(3-nitrophenyl) morpholin-2-++ +ylmethyl]phosphinic acid (CGP 71978) (pA2 = 6.5 +/- 0.05) > cyclohexylmethyl-[(2R,5S)-5-phenyl-morpholin-2-ylmethyl++ +]phosphinic acid (CGP 71980) (pA2 = 6.3 +/- 0.15) > its enantiomer cyclohexylmethyl-[(2S,5R)-5-phenyl morpholin-2-ylmethyl++ +]phosphinic acid (CGP 71979) (pA2 = 5.8 +/- 0.1). An open chain analogue of CGP 76290A, CGP 56999A (3-[1(R)-[(3-cyclohexylmethyl hydroxyphosphinoyl)-2(S)-hydro xypropyl-amino]-ethyl]benzoic acid lithium salt) gave a pA2 of 6.6 +/- 0.2. In GABA(B) receptor binding assays, CGP 71982 (the racemic mixture of CGP 76290A and CGP 76291A), CGP 76290A, CGP 76291A, CGP 71978, CGP 71980 and CGP 71979 had IC50 values against [3H]CGP 27492 binding of 8, 1.85, 69, 124, 326 and 1460 nM, respectively. In electrically-evoked [3H]GABA release from rat cortical slices, CGP 71982, CGP 71978, CGP 71980 and its enantiomer CGP 71979, antagonised GABA(B) autoreceptors with EC150 values of 2.5, 33, 181 and 474 nM, respectively. These compounds form a novel class of potent GABA(B) receptor antagonists. PMID- 9865527 TI - The morpholino-acetic acid analogue Sch 50911 is a selective GABA(B) receptor antagonist in rat neocortical slices. AB - The pharmacological properties of (+)-(S)-5,5-dimethylmorpholinyl-2-acetic acid (Sch 50911) were evaluated on GABA(B) receptors in rat neocortical slices. The GABA(B) receptor agonist, baclofen, produced a concentration-dependent depression of the frequency of spontaneous discharges in slices maintained in Mg2+-free Krebs medium with an EC50 of 6 microM, reversibly antagonised by Sch 50911 (5, 10 and 25 microM) with an apparent pA2 of 6.0 +/- 0.1. The (-) enantiomer Sch 50910 (500 microM) and the racemic des-methyl analogue Sch 48588 (500 microM) were inactive. In slices preloaded with [3H]GABA, Sch 50911 antagonised GABA(B) autoreceptors, increasing the electrically-stimulated 3H overflow in a concentration-dependent manner, with an IC50 of 3 microM. The maximal effect (148 +/- 10.5%) was found at 10 microM, but at 50 microM the response was reduced to 67 +/- 19%. In contrast, evoked release was unaffected by Sch 50910 (100 microM) whilst Sch 48588 at 100 microM increased the overflow by 51.3 +/- 11.6%. In summary, Sch 50911 is a relatively potent antagonist of considerable potential in studies of GABA(B) receptor function. PMID- 9865528 TI - The effects of 5-HT1A, 5-HT1B and 5-HT1D receptor agonists on trigeminal nociceptive neurotransmission in anaesthetized rats. AB - Pre-clinical studies have suggested that one mechanism of antimigraine action of the 'triptan' 5-HT1B/1D receptor agonists may be through inhibition of central nociceptive transmission in the trigeminal dorsal horn. In anaesthetized rats, the 5-HT1B/1D receptor agonist, zolmitriptan (up to 3 mg kg(-1), i.v.), inhibited the action potential discharge of single trigeminal neurones to noxious electrical stimulation of the middle meningeal artery. In contrast, the selective 5-HT1B receptor agonist, CP-93,129 (3-(1,2,5,6-tetrahydropyrid-4-yl)pyrrolo[3,2 b]pyrid-5-one), and the 5-HT1A receptor selective agonist 8-hydroxy-2-(di-n propylamino)-tetralin (8-OH-DPAT) had no effect in this assay at up to 3 mg kg( 1), i.v.. Brain penetrant, triptan 5-HT1B/1D receptor agonists may therefore mediate their central trigeminal anti-nociceptive action in the rat via 5-HT1D, but not 5-HT1B or 5-HT1A, receptors. PMID- 9865529 TI - Possible involvement of protein kinase C in the attenuation of the morphine induced Straub tail reaction in diabetic mice. AB - To investigate the role of protein kinase C in the attenuation of the morphine induced Straub tail reaction in diabetic mice, we examined the effects of protein kinase C activator or inhibitor on the i.c.v. morphine-induced Straub tail reaction in mice. This reaction was less in diabetic mice than in normal mice. Intracerebroventricular pretreatment with phorbol 12,13-dibutyrate (50 pmol), a potent protein kinase C activator, attenuated the morphine-induced Straub tail reaction in normal mice, but not in diabetic mice. I.c.v. pretreatment with calphostin C (10 pmol), a selective protein kinase C inhibitor, enhanced the reaction in diabetic mice, but not in normal mice. The dose-response curve for the morphine-induced Straub tail reaction in normal mice, but not in diabetic mice, was shifted to the right by i.c.v. pretreatment with phorbol 12,13 dibutyrate (50 pmol). Furthermore, i.c.v. pretreatment with calphostin C (3 pmol) shifted the dose-response curve to the left in diabetic mice, but not in normal mice. These results indicate that activation of protein kinase C reduces the morphine-induced Straub tail reaction in normal mice. Also, the attenuation of the morphine-induced Straub tail reaction in diabetic mice may be due in part to increased protein kinase C activity. PMID- 9865530 TI - Role of histamine H1 and H2 receptor antagonists in the prevention of intimal thickening. AB - Vascular smooth muscle cell migration to the intima from the media and proliferation in the intima play key roles in atherosclerosis and restenosis after coronary angioplasty. Histamine released from adherent platelets at the injured artery and from mast cells in atheromas has stimulant actions on both smooth muscle cell migration and proliferation, and histamine receptor antagonists abolish the effect of histamine in vitro. The aim of this study was to examine the effect of histamine receptor antagonists on intimal thickening. Endothelial injury in the mouse femoral artery was induced by a photochemical reaction between localized irradiation by green light and intravenously administered rose bengal. The histamine H1 receptor antagonist, diphenhydramine, at a dose of 30 mg/kg or the histamine H2 receptor antagonist, cimetidine, at a dose of 200 mg/kg was intraperitoneally administered to mice for 21 days after endothelial injury. Twenty-one days after endothelial injury, morphometric analysis was performed to measure the cross-sectional areas of the intima and media. Diphenhydramine significantly reduced the intimal area to 1.1 +/- 0.3 (x 10(-3) mm2) compared with the value in the control group, which was 6.2 +/- 1.4 (x 10(-3) mm2), but cimetidine (5.5 +/- 1.9, x 10(-3) mm2) did not. Similarly, the ratio of intimal area to medial area in the diphenhydramine-treated group but not in the cimetidine-treated group was significantly reduced (83%). In the in vitro study, cimetidine inhibited neither proliferation nor migration of mouse vascular smooth muscle cells stimulated by platelet-derived growth factor (PDGF). In contrast, diphenhydramine significantly (P < 0.05) inhibited proliferation in a dose-dependent manner, but did not inhibit migration. These results suggest that diphenhydramine, a histamine H1 receptor antagonist, reduced the formation of intimal hyperplasia, at least in part due to inhibition of cell proliferation. However, cimetidine, a histamine H2 receptor antagonist, was ineffective. Histamine may play a key role in intimal thickening, in part via histamine H1 receptors in this model. PMID- 9865531 TI - Effect of moxonidine on blood pressure and sympathetic tone in conscious spontaneously hypertensive rats. AB - The effects of moxonidine on blood pressure, heart rate and sympathetic tone were studied in conscious spontaneously hypertensive rats. Intravenous moxonidine (80 nmol) transiently increased blood pressure without affecting heart rate or splanchnic nerve activity. Moxonidine (20-80 nmol) given into the fourth cerebral ventricle dose-dependently lowered mean arterial pressure, heart rate and sympathetic outflow (maximally by 60 +/- 3 mm Hg, 148 +/- 10 beats min(-1) and 15 +/- 3 microV). Moxonidine was more effective by this route than after the injection into the lateral ventricle. Clonidine (20-80 nmol) produced an initial pressor response after both intracerebroventricular routes of administration. A decrease in blood pressure was observed only when clonidine was given into the fourth ventricle. Clonidine decreased heart rate and splanchnic nerve activity similarly like moxonidine when the substances were given into the fourth ventricle. The data imply that the hypotensive effect of moxonidine is related to central sympathoinhibition. The main site of this action appears to be in the brainstem region. PMID- 9865532 TI - The canine external carotid vasoconstrictor 5-HT1 receptor: blockade by 5-HT1B (SB224289), but not by 5-HT1D (BRL15572) receptor antagonists. AB - In vagosympathectomised dogs pre-treated intravenously (i.v.) with mesulergine (300 microg/kg), 1-min intracarotid (i.c.) infusions of 5-hydroxytryptamine (5 HT; 0.3-30 microg/min) and sumatriptan (1-30 microg/min) dose-dependently decreased external carotid blood flow, without affecting mean blood pressure or heart rate. Treatment with the selective 5-HT1B receptor antagonist SB224289 (2,3,6,7-tetrahydro-1'-methyl-5-[2'-methyl-4'(5-methyl-1,2,4-oxadiazo l-3-yl) biphenyl-4-carbonyl]furo[2,3f]indole-3-spiro-4'-piperidine hydrochloride; 30-300 microg/kg, i.v.) produced a potent, specific and dose-dependent blockade of this response, whereas the selective 5-HT1D receptor antagonist BRL15572 (1-(3 chlorophenyl)-4-[3,3-diphenyl(2-(S,R) hydroxypropanyl)piperazine]hydrochloride; 30-300 microg/kg, i.v.) was ineffective. It is concluded that mainly 5-HT1B, but not 5-HT1D receptors mediate the canine external carotid vasoconstriction by 5-HT and sumatriptan. PMID- 9865533 TI - 17Beta-estradiol acutely improves endothelium-dependent relaxation to bradykinin in isolated human coronary arteries. AB - Improvement of endothelial function has been implicated in the cardioprotective effects of estrogens in women. In isolated human coronary arteries, we investigated whether 17beta-estradiol affects endothelium-dependent responses to bradykinin, an endothelium-derived vasodilator locally produced by endothelial cells. Concentration-response curves to bradykinin (0.03-300 nM) or nitroglycerine (0.01-1 microM) were obtained before and after 30 min of incubation with 17beta-estradiol (3 microM) or solvent control (ethanol 0.2% vol/vol). Incubation with 17beta-estradiol enhanced relaxations to bradykinin (from 43 +/- 6 to 83 +/- 3%, P < 0.0001) but not those to nitroglycerine (n.s.). Improvement of bradykinin-mediated endothelium-dependent relaxation may represent a novel mechanism contributing to the cardioprotective effects of estrogen in women. PMID- 9865534 TI - Hydralazine prevents endothelial dysfunction, but not the increase in superoxide production in nitric oxide-deficient hypertension. AB - Dilator responses, superoxide anion-production, endothelial nitric oxide (NO) synthase and soluble guanylyl cyclase expression were determined in aortic rings from Wistar rats treated for 5 weeks either with the NO synthase inhibitor NG nitro-L-arginine-methylester (L-NAME), L-NAME plus hydralazine or placebo. In the L-NAME-treated group, acetylcholine-induced relaxation was significantly attenuated whereas it was nearly normal in the L-NAME/hydralazine group. This difference was even more pronounced following inhibition of the endogenous superoxide dismutase using diethyldithiocarbamate. Aortic superoxide production was significantly elevated in both L-NAME-treated groups and hydralazine had no acute effect on superoxide formation. Expression of endothelial NO synthase was similar in all three groups whereas the attenuated soluble guanylyl cyclase expression in rats treated with L-NAME was nearly normalised by concomitant hydralazine treatment. These results demonstrate that in NO-deficient hypertension hydralazine treatment improves vasodilator responses but not the increased superoxide production. PMID- 9865535 TI - Ba2+ selectively inhibits receptor-mediated contraction of the esophageal muscularis mucosae. AB - The aim of the present study was to examine the effect of Ba2+ on acetylcholine- and KCl-induced contractions of the guinea-pig esophageal muscularis mucosae. When the muscularis mucosae was pretreated with nicardipine (1 microM), Ba2+ (0.1 30 mM) markedly inhibited the acetylcholine (3 microM)-induced tone, and at 10-30 mM the tone returned to its basal level. In contrast, Ba2+ (0.1-30 mM) slightly increased the KCl (60 mM)-induced tone. Moreover, the Ba2+ (30 mM)-increased KCl tone was completely inhibited by treatment with nicardipine (0.3-1 microM). In conclusion, Ba2+ both selectively inhibits receptor-mediated contraction of the muscularis mucosae and itself permeates through voltage-dependent Ca2+ channels. PMID- 9865536 TI - Induction of estradiol-2-hydroxylase and ethoxyresorufin-O-deethylase by 3 substituted indole compounds. AB - Estrogen can be hydroxylated at both 2- and 16alpha-positions. These two reactions are mutually exclusive. The 2-hydroxylated estrogen is relatively inactive compared with the 16alpha-derivative; hence, one approach in anti estrogenic therapy is to look for drugs that can induce the 2-hydroxylation pathway. In the present study, using Balb/c and C57B/6 mice as the animal models, the induction effect of several isoprenyl compounds on estradiol-2-hydroxylase and ethoxyresorufin-O-deethylase activities was studied. The compounds examined included 2'- and 3'-methylbutadienyl-indoles and their respective acid condensation products, isopropyl indolocarbazole and yuehchukene; positional isomers of indole carbinols and carboxyaldehydes, as well as 3 methylcholanthrene, the prototype inducer of cytochrome P450 1A1. Our results demonstrated that while all of them were capable of inducing cytochrome P450 1A1 mediated ethoxyresorufin-O-deethylase activity, only the 3' isomers could induce estradiol-2-hydroxylase activity. The induction of these two activities did not show any direct correlation, suggesting that cytochrome P450 1A1 was not the same enzyme catalyzing both ethoxyresorufin-O-deethylation and estradiol-2 hydroxylation. Nevertheless, both inductions were mediated by the aryl hydrocarbon receptor. Among the compounds tested, yuehchukene showed competitive binding to estrogen receptor. This, together with the induction of estradiol-2 hydroxylase activity, may account for the anti-estrogenic effect of yuehchukene. PMID- 9865537 TI - Carbachol, an acetylcholine receptor agonist, enhances production in rat aorta of 2-arachidonoyl glycerol, a hypotensive endocannabinoid. AB - The production of 2-arachidonoyl glycerol, an endogenous cannabinoid, is enhanced in normal, but not in endothelium-denuded rat aorta on stimulation with carbachol, an acetylcholine receptor agonist. 2-Arachidonoyl glycerol potently reduces blood pressure in rats and may represent an endothelium-derived hypotensive factor. PMID- 9865538 TI - Hippocampal sclerosis revisited. AB - Studies dating back more than 150 years reported a relationship between hippocampal sclerosis and epilepsy. Retrospective studies of patients who underwent temporal lobectomy for intractable partial epilepsy found a relationship between a history of early childhood convulsions, hippocampal sclerosis, and the development of temporal lobe epilepsy. Many believe that febrile seizures lead to hippocampal damage and this in turn predisposes the patient to the development of temporal lobe epilepsy. Studies in adult rats have shown that seizures can lead to hippocampal damage and unprovoked recurrent seizures. However, many questions remain as to the relevance of early childhood seizures to hippocampal sclerosis and temporal lobe epilepsy. Human prospective epidemiologic studies have not shown a relationship between early childhood seizures and temporal lobe epilepsy. Recent MRI studies in humans suggest that a preexisting hippocampal lesion may predispose infants to experience febrile seizures, later on hippocampal sclerosis, and possibly temporal lobe epilepsy may occur. Unlike the studies in adult rats, normal immature rats with seizures have not been shown to develop hippocampal damage or unprovoked seizures in adulthood. Furthermore, animal studies reveal that preexisting brain abnormalities can predispose to hippocampal damage following seizures early in life. This paper reviews evidence for and against the view that early childhood convulsions, hippocampal sclerosis, and temporal lobe epilepsy are related, while also exploring clinical and animal studies on how seizures can lead to hippocampal damage, and how this can result in temporal lobe epilepsy. By better understanding the cause and effect relationship between early childhood seizures and hippocampal injury in normal and abnormal brains specific treatments can be developed that target the pathogenesis of epilepsy. PMID- 9865539 TI - Cerebral oxygenation and hemodynamics during hyperventilation and sleep in patients with Rett syndrome. AB - We continuously monitored changes in cerebral oxygenation and hemodynamics in the frontal lobes of six patients with Rett syndrome during the awake state, which is associated with hyperventilation (HV) and breath-holding (BH), and during sleep by near-infrared spectroscopy. We also monitored three adult volunteers during simulated episodes of HV and BH. In patients with Rett syndrome, the oxygenated hemoglobin (HbO2) and total hemoglobin (HbT) decreased significantly during HV and BH in the awake state compared with the sleep state. The HbO2 and HbT decreased gradually in adult volunteers in response to prolonged episodes of HV and BH. Further studies are required to investigate the relationship, if any, between the discovered continuous decreases in HbO2 and HbT during the awake state and the brain damage seen in patients with Rett syndrome. PMID- 9865540 TI - Polysomnographic studies of Lesch-Nyhan syndrome. AB - Three cases of Lesch-Nyhan syndrome (LNS) were examined by polysomnography to assess the brainstem function, and to determine the causes of the neurological manifestations and sudden death in this syndrome. In the two older cases, the amount of slow wave and rapid eye movement (REM) sleep, the REM density and the frequency of REM bursts were decreased. In the youngest case, symmetrical phasic movements of all four limbs were observed at all sleep stages other than REM sleep. Although movements other than these symmetrical body movements appeared to be normal in this case, the frequency of twitch movements showed an abnormal pattern in each sleep stage in the two older cases. These findings suggest that in the brainstems of younger cases with LNS the REM-non REM generator as well as multiple neurotransmitter systems influencing body movements during sleep remain relatively normal, but become progressively impaired in adult cases. Severe obstructive apnea was observed in one case with hypothyroidism, but there were no respiratory abnormalities in other two cases. PMID- 9865541 TI - Ultrastructural study of enteric ganglia in three patients with Rett syndrome. AB - In order to verify whether a pseudo-obstruction syndrome was associated with morphological changes in enteric ganglia, we performed an ultrastructural study on rectal biopsy specimens in three patients with Rett syndrome. Features of enteric neurons, detected to a different extent in all three biopsy specimens, included an abnormal dilatation of endoplasmic reticulum with a disorganization of cisternae of the Golgi apparatus, and masses of unidentified electron-dense granulo-filamentous material, probably of lipidic origin, observed in the perikaryon. Large electron-lucent membrane-bound vacuoles were found mostly within satellite glial cells. Sometimes, the axon terminals were swollen and showed intraxonal vacuolization. We conclude that the reported findings do not represent a specific sign of degeneration and do not constitute a significant morphological marker of disease. PMID- 9865542 TI - Cognitive improvement despite minimal arachnoid cyst decompression. AB - There are relatively few reports that evaluate the cognitive functions of patients with arachnoid cysts. Presumably, these 'silent cysts' are regarded as incidental findings with no functional significance. Although postoperative clinical improvement is well documented in patients with significant reduction in cystic volume, the current report describes a patient who underwent cystoperitoneal shunting due to mass effect, with minimal postoperative decompression. Neuropsychological testing indicated significant cognitive improvement in verbal learning, memory, visual-perceptual abilities, constructional skills, conceptual shifting, and psychomotor speed after shunt placement, despite marginal evidence of decompression. These findings suggest that (1) significant cognitive changes can occur in these patients, despite minimal postoperative regression of the lesion, (2) cognitive measures may provide an alternative, functional index of outcome efficacy, and (3) reliance on traditional outcome measures (i.e. anatomical decompression or resolution of clinical symptoms) may underestimate the efficacy of surgical intervention for these patients. PMID- 9865543 TI - A case of giant axonal neuropathy showing focal aggregation and hypophosphorylation of intermediate filaments. AB - We report here the clinicopathological features of a typical case of giant axonal neuropathy (GAN). Scanning electron microscopy of the hair of this case revealed an extraordinarily irregular cuticle. Focal accumulation of intermediate filaments in axons, Schwann cells, muscle fibers and skin fibroblasts were also found under an electron microscopy. When examined immunocytochemically, muscle fibers exhibited local disruption of the filamentous network in the subsarcolemmal space and in the central cytoplasm accompanied by focal accumulation of desmin. The intracellular network of vimentin was also disrupted, exhibiting global accumulation in some of the cultured skin fibroblasts. Decreased interneurofilament spacing was found in enlarged axons, suggesting the presence of hypophosphorylation of neurofilaments in this patient. These findings suggest general disorganization, abnormal distribution and possible defective phosphorylation of intermediate filaments in GAN. PMID- 9865545 TI - Simple methods for assessing urinary iodine, including preliminary description of a new rapid technique ("Fast B"). AB - The urinary iodine is the best laboratory measure of iodine nutrition. Several simple methods are suitable for developing countries. The present article reviews these briefly. It also presents preliminary information on "Fast B", a rapid method that puts samples into convenient ranges rather than giving absolute values for individuals. This grouping by ranges is satisfactory for many epidemiological purposes and is considerably more rapid than other approaches. The choice among methods depends on the intended application, the number of samples, the cost and the technical capability. Having several different methods available allows the user to select the one best suited to specific needs. Further development of simple methods and appropriate combinations of steps from different ones deserve encouragement as a means of improving monitoring in the campaign to achieve sustainable elimination of iodine deficiency. PMID- 9865544 TI - Implications of the molecular characterization of the sodium-iodide symporter (NIS). AB - The recently cloned sodium-iodide symporter (NIS) represents a key molecule for thyroid function by efficiently accumulating iodide from the circulation into the thyrocyte against an electrochemical gradient. This uptake requires energy, is coupled to the action of Na+/K+-ATPase, and stimulated by TSH, the main hormone regulating thyroid-specific functions. NIS mutations are found in congenital hypothyroidism, and potential defects in the NIS gene, its expression, or function of the NIS protein are currently under investigation in various thyroid diseases. Increased NIS expression has been found in autonomous adenoma and Graves' disease, decreased levels of NIS protein and/or mRNA were observed in Hashimoto's disease, cold nodules, most thyroid cancers and cell lines derived therefrom. Autoantibodies directed against NIS have been identified in autoimmune thyroid disease and blocking antibodies isolated from sera of patients with Hashimoto's disease inhibit NIS function in NIS-transfected CHO cells. NIS mRNA expression can be up-regulated by retinoic acid in human thyroid carcinoma cell lines whereas retinoic acid treatment decreases NIS expression and function in differentiated rat thyroid FRTL-5 cells. Apart from thyrocytes, NIS is also expressed in other tissues known to transiently accumulate radioiodide, albeit at much lower levels, requiring RT-PCR for detection of the transcript. Diagnostic and therapeutic implications of the recent cloning of the human NIS gene such as development of NIS-directed drugs, ligands, antibodies, vaccines, gene therapeutic approaches combining NIS targeting and expression together with the long-established, efficient and safe method of radioiodide therapy are discussed both for application to thyroid related diseases and carcinoma, and non-thyroid benign and malignant diseases. Apart from these therapeutic and diagnostic perspectives the availability of the NIS gene will also open new opportunities to develop sensitive and homologous diagnostic test systems to identify factors involved in autoimmune thyroid disease, evolution of goitre, adenoma and thyroid cancer as well as NIS-directed new drugs. Advanced and sophisticated molecular diagnostic approaches (RT-PCR from fine needle aspirations, screening for mutations, analysis of gene defects) are already developed for NIS and will complement or overcome some established procedures in thyroid diagnostics. PMID- 9865546 TI - Rapid urinary iodide test. AB - Assessment of iodine deficiency and monitoring of iodine supplementation programs demands rapid, simple and cost-effective methods for the determination of urinary iodide concentrations. We propose a rapid test based on the iodide-catalyzed oxidation of 3,3',5,5'-tetramethylbenzidine by peracetic acid/H2O2 to yield colored products. The color of the chemical reaction is compared with color categories of a pictogram corresponding to three ranges (<10, 10-30, and >>30 microg/100 mL) of iodide concentrations. The test is very easy to perform and does not require any instrumentation or apparatus. Sample preparation is simple and consists in the removal of interfering substances by disposable columns, 65 x 10.5 mm, packed with specifically prepared activated charcoal. For comparison with a reference method for measuring urinary iodide (HPLC), we determined the iodide concentrations of 370 random (untimed) urine samples from consecutive patients by both HPLC and the rapid test. The results obtained by both methods are in close agreement with respect to classification of the samples according to the above three ranges, with a maximum difference of <5% for each range. This rapid test is therefore very well suited to epidemiological surveys of iodine deficiency especially in developing countries. PMID- 9865547 TI - Evaluation of the iodine intake: problems of the iodine/creatinine ratio- comparison with iodine excretion and daily fluctuations of iodine concentration. PMID- 9865548 TI - Iodine supplementation during pregnancy: importance and biochemical assessment. PMID- 9865549 TI - Ammonium persulfate: a new and safe method for measuring urinary iodine by ammonium persulfate oxidation. AB - The chloric acid method is most commonly used to obtain accurate and reproducible measurements of iodine and removes interfering substances. Unfortunately, chloric acid is a potential hazard requiring an explosion proof hood among other precautions. We have developed a simple, convenient, and economic method for measuring urinary iodine using 1 mol/L ammonium persulfate, a non-explosive, non hazardous chemical, as the oxidizing reagent. The oxidation procedure can be completed in 30 minutes at a temperature of 91-95 degrees C. The iodine in the urine is then measured by a modification of the traditional colorimetric method of Sandell-Kolthoff. 110 urine samples collected from a mixed population of healthy males and females, ranging in age from 6 to 79 years and living in the United States were analyzed for iodine content by two methods: the proposed ammonium persulfate method and the chloric acid method. The ammonium persulfate method has an intra assay CV of 9.1% at 0.42 +/- 0.04 micromol/L (mean +/- SD), 7.8% at 1.46 +/- 0.11 micromol/L and 4.0% at 3.54 +/- 0.14 micromol/L. The inter assay CV is 10.2% at 0.46 +/- 0.05 micromol/L and 7.9% at 3.27 +/- 0.26 micromol/L. Recovery of iodine added to urine in vitro was 107%, 94% and 97% for 0.42 micromol/L, 0.77 micromol/L and 3.64 micromol/L, respectively. The lower limit of detectability was 0.0034 microgI. Values for iodine in 110 urines measured by the reference chloric acid method ranged from 0.06 to 8.03 micromol/L and by the ammonium persulfate method from 0.05 to 7.4 micromol/L. The persulfate method (y) correlated extremely closely with the reference chloric acid method (x) by the Pearson correlation (y = 0.923x + 0.810 micromol/L, and r = 0.994, Syx = 1.841). In conclusion a new, safe, simple method for measuring urinary iodine is described which uses ammonium persulfate as the oxidizing agent for the removal of interfering substances. PMID- 9865550 TI - Methodological and analytical aspects of simple methods for measuring iodine in urine. Comparison with HPLC and Technicon Autoanalyzer II. AB - This work describes an optimization of a simple photometric determination of iodine concentrations in urine using a modified ceric arsenite method with ammonium persulfate as oxidant. By means of this sensitive method iodine concentrations can be determined in very small specimens (50 microL). Urine samples (105) collected from a mixed population, were analyzed for urine iodine content by the optimized ammonium persulfate method, a Technicon Autoanalyzer II and a paired-ion-RP HPLC. We found that the precision of this optimized ammonium persulfate method yields inter assay CVs of <10% for urinary iodine concentrations >10 microg/dL. Recovery of [123I]iodide added to urine in vitro was 100.9 +/- 2.4%. The detection limit was 0.0029 microg iodine. There was a high correlation between all three methods (r > 0.94 in any case) and the interpretation of the results was consistent. We conclude that this simple, manual ammonium persulfate method is suitable for urinary iodine analysis and can be performed in any routine clinical laboratory. PMID- 9865551 TI - X-ray fluorescence analysis (XFA) of thyroidal iodine content (TIC) with an improved measuring system. AB - X-ray fluorescence analysis is based on the principal that the electron structure of stable iodine in the thyroid is excited by Americium-241 gamma rays to emit a characteristic fluorescence radiation which is proportional to the amount of iodine present in the gland. A stationary measuring system consisting of a 11.1 GBq Am-241 source and a high-purity Germanium detector with spectrum analyser has been improved by a PC guided method for sonographic definition of the measuring volume. The lower limit of detectibility of the system corresponds to 0.01 mg of Iodine per ml of thyroid volume; the in vivo precision given as coefficient of variation amounts to 15%. The thyroid is exposed with a radiation dose of 6 microSv per measurement. First studies with this improved system carried out in 50 female volunteers between 20 and 40 years of age with normal thyroid volumes resulted in a mean iodine concentration of the thyroid of 0.665 +/- 0.304 mg/ml. The mean iodine excretion in urine was normal with 10.8 +/- 10.4 microg/dl. PMID- 9865552 TI - Kinetics of acute and chronic iodine excess. AB - Iodine plays an important role in thyroid physiology resulting from its importance as a requisite substrate for the synthesis of thyroid hormones and from its action as a regulator of thyroid function. Following intestinal absorption, inorganic iodide is largely confined to the extracellular fluid. Serum concentrations of inorganic iodide well reflect the amount of iodine present in the extracellular compartment. Since serum inorganic iodide levels are important determinants of thyroid iodine uptake, serum iodide measurement offers a valuable tool for the investigation of many basic and clinical aspects of thyroid iodine metabolism. Here we summarize important aspects of iodine metabolism and focus selectively on technical aspects of serum inorganic iodide measurement and on the kinetics of inorganic iodide in various states of iodine excess. Presently, paired-ion, reversed-phase HPLC with electrochemical detection is obviously the best method for measurement of serum inorganic iodide being highly sensitive, easy to perform, and almost completely insensitive to interfering substances. Using this method, we could demonstrate an acute increase of serum inorganic iodide during the administration of large amounts of iodide as Lugol's solution given preoperatively in patients with Graves' hyperthyroidism. In patients under treatment with the iodine containing drug amiodarone (n=37), serum inorganic iodide levels were highly elevated (range 3.5-208.2 microg/dl, median 36.6 microg/dl). Serum concentrations of inorganic iodide were correlated neither to the daily amiodarone dose, nor to the serum levels of amiodarone. PMID- 9865553 TI - Iodine intake and thyroid carcinoma--a potential risk factor. AB - The role of enhanced thyroid-stimulating hormone (TSH) secretion, in the aetiology of thyroid cancer is not totally consistent. Circumstances and conditions which cause (e.g., iodine deficiency, through suboptimal intake in water and food) or indicate (e.g., goitre) increased TSH secretion have been associated to increased risk of thyroid cancer, most notably follicular and anaplastic carcinomas. Elevated incidence and mortality rates of thyroid cancer, however, are also found in areas were iodine intake is high (Hawaii, Iceland). At least in some countries (Switzerland), a favourable impact of the introduction of iodized salt on mortality from thyroid cancer has been reported. Elsewhere, the correction of iodine deficiency has coincided with elevations of diagnostic standards (e.g., spread of thyroid scintigraphy, ultrasound, and fine-needle biopsy) and corresponding increases in incidence of papillary carcinomas, often clinically silent, thus hampering a distinction of the two phenomena. Upward trends of papillary carcinoma incidence have, however, been seen in most affluent countries, irrespective of the iodine status of the population. PMID- 9865554 TI - Diagnosis of gastric cancer up to three years after negative upper gastrointestinal endoscopy. AB - BACKGROUND AND STUDY AIMS: The degree of accuracy of gastroscopy for the detection of gastric cancer is poorly understood. The aim of this retrospective study was to determine the accuracy of gastroscopy by using cancer registry records. PATIENTS AND METHODS: Gastroscopic examinations (n = 37094) conducted between 1984 and 1989 were studied by linking them with hospital-based and population-based (Fukui Prefecture, Japan) cancer registry records between 1984 and 1992. False-negative gastroscopies that had been carried out within the three years preceding the diagnosis of gastric cancer were identified. RESULTS: The numbers of true-positive, false-positive, and false-negative examinations carried out were 659, six and 155, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value were 81.0%, 100.0%, 99.1%, and 99.6%, respectively. The overall diagnostic accuracy of gastroscopy was 99.6%. There was little difference in sensitivity results between the patient groups with regard to reason for referral, type of endoscope used, experience of endoscopist, or location of gastric cancer. The percentage of tumours representing early gastric cancer, identified after false-negative gastroscopy, was lower for those situated in the cardia or gastric body than for those in the angular notch or the antrum. CONCLUSIONS: The accuracy of gastroscopy in the detection of gastric cancer is satisfactory, but false-negative results are sometimes obtained. We emphasize the importance of repeated endoscopic examination for the detection of gastric cancer. PMID- 9865555 TI - Colonoscopic evaluation of severe hematochezia in an Oriental population. AB - BACKGROUND AND STUDY AIMS: Hematochezia is a common clinical problem. When the bleeding is brisk and continuous it requires prompt hospital admission and careful diagnostic evaluation and management. Colonoscopy has become the first line investigative modality in patients presenting with severe hematochezia in many centers, including ours. A retrospective review was carried out to evaluate the effectiveness of colonoscopy in determining the cause of severe hematochezia in our Oriental population. PATIENTS AND METHODS: One hundred and ninety patients with severe hematochezia underwent colonoscopy at the National University Hospital, Singapore, from 1 January 1988 to 31 December 1994. Their records were retrieved and the data analyzed for sex, age, presentation, concomitant medical conditions, prevalence of recent non-steroidal anti-inflammatory drugs ingestion, past history of hematochezia, investigations, subsequent interventions and outcome. RESULTS: Colonoscopy as the fist-line investigative modality identified the site and cause of hematochezia in 78% (148/190) of cases. The site of bleeding remained "obscure" even after additional investigations in 15% (29/190) of cases. The commonest cause of severe hematochezia in our Oriental population was diverticular disease (30%, 57/190) with right-sided diverticular bleeding constituting 44% (25/57) of these cases. Overall, bleeding stopped spontaneously in 81% (154/190) of cases. Surgery was performed in 16% (30/190) of cases. The mortality related to severe hematochezia in this series was 5% (9/190). CONCLUSIONS: The diagnostic efficiency of colonoscopy in defining the site and cause of severe hematochezia in the Oriental population is comparable to most Western series. The commonest cause of severe hematochezia in our population was diverticular disease. PMID- 9865556 TI - A prospective randomized trial of Teflon versus polyethylene stents for distal malignant biliary obstruction. AB - BACKGROUND AND STUDY AIMS: Clogging of biliary stents continues to be a major clinical problem. Different polymer materials may have different effects on clogging. In vitro studies have shown a direct relation between the frictional coefficient of a polymer and the amount of encrusted material. Teflon appeared to be the best polymer for biliary stents. Two different types of stents made of Teflon have been tested in clinical practice and showed favourable patency rates. However, a randomized trial has never been performed. We compared the patency of an Amsterdam-type polyethylene stent with a Teflon stent in a prospective randomized trial. PATIENTS AND METHODS: Between September 1995 and November 1996, 42 patients received a Teflon stent and 42 patients a polyethylene stent. All patients had a distal malignant biliary stricture without a previous drainage procedure. Diagnoses included carcinoma of the pancreas (n = 76), papilla (n = 1), bile duct (n = 5) and metastases (n = 2). The internal and external diameter (10 Fr), length (9 cm) and stent design (a straight stent with two side flaps and one side hole at each end) were similar for both stents. RESULTS: A reduction in bilirubin of more than 20% within one week was seen in 91% of the patients. Early complication rates were similar in both groups (10%). The median follow-up was 142 days. Stent dysfunction occurred in 28 Teflon and 29 polyethylene stents. The thirty-day mortality was 14% in both groups. Patient survival did not differ significantly between the groups (median survival: Teflon 165 days, polyethylene 140 days). The median stent patency was 83 days for Teflon and 80 days for polyethylene stents, and was not significantly different either. CONCLUSION: Teflon material did not improve patency in biliary stents with an Amsterdam-type design. PMID- 9865557 TI - Feasibility of stent placement above the sphincter of Oddi ("inside-stent") for patients with malignant biliary obstruction. AB - BACKGROUND AND STUDY AIMS: Animal studies suggest that placement of endobiliary stents wholly above the papilla (the "inside-stent") prolongs the duration of stent patency. We reviewed the cholangiographic characteristics of patients with malignant obstructive jaundice in order to evaluate the feasibility of this approach and to assist in the design of a new stent. PATIENTS AND METHODS: A review was conducted of 270 patients who had undergone endoscopic retrograde cholangiopancreatography (ERCP) for malignant obstructive jaundice at our hospital, to determine the relative frequencies of the types of cancers. The ERCP films of 59 relevant consecutive patients with malignant obstructive jaundice were studied to determine the length of the stricture, the distance between the stricture and the papilla, and the angulation of the common duct. RESULTS: The most frequent primary cancers causing malignant obstructive jaundice were pancreatic cancer (57%), biliary cancer involving the hilum (19%, including metastatic disease), nonhilar biliary cancer (14%), and papillary cancer (10%). The length between the lower end of the stricture and the sphincter was more than 2 cm for all hilar biliary cancer, representing two-thirds of nonhilar biliary cancer cases, while it was less than 1 cm for most pancreatic cancers. There was an average angulation of 30-40 degrees in the axis of the distal common bile duct. CONCLUSIONS: For most biliary cancers, the stricture is high enough to allow the placement of a stent fully above the papilla, but this is not possible for most pancreatic cancers (or any papillary cancers). An "inside-stent" approach would be possible in about one-third of patients with malignant obstructive jaundice if a clearance length of over 2 cm between the stricture and the sphincter is required, and in about 45% of patients if a minimum clearance of 1 cm is required. A 30-40 degrees flexion in the distal part of the stent would approximate the average angulation in the axis of the common bile duct and help to prevent dislocation. PMID- 9865558 TI - Needle-knife papillotomy: a helpful and safe adjunct to endoscopic retrograde cholangiopancreatography in a selected population. AB - BACKGROUND AND STUDY AIMS: Needle-knife papillotomy (NKP) has been shown to be a useful alternative when standard techniques fail to provide access during ERCP. A retrospective review was conducted to reevaluate the indications, efficacy, and complications of NKP at a tertiary referral center. PATIENTS AND METHODS: A total of 1205 therapeutic endoscopic retrograde cholangiography procedures (ERCPs) were reviewed. Sixty-eight patients (5.6%) had undergone NKP after an established algorithm of techniques had failed to provide access. The NKP results were analyzed in two periods, including initial experience with 470 ERCPs (group 1, NKP n = 22) and later experience with 735 patients (group 2, NKP n = 46). RESULTS: Immediate free cannulation in group 1 was achieved in 14 of 22 patients (64%), vs. 34 of 46 (74%) in group 2. The delayed cannulation rate was five of eight patients in group 1 (62.5%) vs. 11 of 12 in group 2 (92%). The success rate was 19 of 22 patients in group 1 (86%) vs. 45 of 46 in group 2 (98%). The overall success rate was 64 of 68 (94%). Successful cannulation led to a therapeutic intervention in 94% of these patients. A complication rate of 6%, without mortality, was noted. CONCLUSIONS: NKP is a valuable tool that allows a high success rate for cannulation, with a low complication rate. The success rate increases with operator experience. NKP should be carried out by experienced endoscopists after standard maneuvers fail to provide access, and when cannulation is likely to be followed by a therapeutic intervention. PMID- 9865559 TI - Routine biliary sphincterotomy may not be indispensable for endoscopic pancreatic sphincterotomy. AB - BACKGROUND AND STUDY AIMS: It is generally accepted that biliary sphincterotomy is mandatory to avoid possible cholestasis and infection due to edema after pancreatic sphincterotomy. However, biliary sphincterotomy is an invasive procedure and the above claim on dual sphincterotomy has not been proven by a prospective randomized study. The aim of our study was to determine whether cholangitis develops more frequently when the patients have not undergone concomitant biliary sphincterotomy during the endoscopic pancreatic sphincterotomy. PATIENTS AND METHODS: From January 1990 to November 1997, 60 patients (38 men, 22 women, age range 19-45 years) with non-alcoholic chronic calcifying pancreatitis were prospectively enrolled. The patients with jaundice (bilirubin > or = 3 mg/dl), cholangitis, or parenchymal liver disease were excluded. The patients were randomly subjected either to dual sphincterotomy (group I, n = 30) or to pancreatic sphincterotomy alone (group II, n = 30). Groups I and II were further classified as IA (or IIA) and IB (or IIB), according to the level of serum alkaline phosphatase (sALP) and the diameter of the common bile duct (CBD). Group IA (or IIA) was defined when abnormal in both sALP (> or = 2 times the upper limit of normal) and CBD diameter (> or = 12 mm), whereas group IB (or IIB) was defined when normal, or solely abnormal in sALP or CBD diameter. RESULTS: As a complication after sphincterotomy, pancreatitis developed in one of eight patients (12.5%) in group IA, whereas cholangitis occurred in one of 22 (4.5%) and hemorrhage in one of 22 (4.5%) cases in group IB. By contrast, in group IIA, the cholangitis developed in 56% (five of nine patients), which was significantly more frequent than in any other groups (P < 0.05). Hemorrhage (one of 21, 4.8%) and pancreatitis (one of 21, 4.8%) occurred in group IIB. CONCLUSIONS: Our results suggest that dual sphincterotomy may be indicated only in patients who have both dilated choledochus and elevated alkaline phosphatase in chronic pancreatitis. Routine biliary sphincterotomy may not be indispensable for pancreatic sphincterotomy. PMID- 9865560 TI - Endoscopic injection of botulinum toxin for biliary sphincter of Oddi dysfunction. AB - BACKGROUND AND STUDY AIMS: Endoscopic sphincterotomy is not without risks, and is also ineffective in about half of patients with type III sphincter of Oddi dysfunction (SOD), i.e. those without clinical evidence of biliary obstruction (normal liver tests, normal bile duct diameter, and regular drainage time at endoscopic retrograde cholangiography). The present study therefore investigated the efficacy and safety of endoscopic botulinum toxin (BTX) injection into the papilla of Vater, and analyzed whether the symptomatic response to BTX injection might be a predictor of outcome for endoscopic sphincterotomy. PATIENTS AND METHODS: Twenty-two patients who had undergone cholecystectomy and had manometrically confirmed type III SOD were enrolled during a three-year study period. All patients received treatment with an endoscopic single-shot injection of 100 mouse units of BTX into the papilla of Vater. Initial symptomatic responses were analyzed six weeks later. If the BTX injection had been ineffective, or if biliary symptoms recurred after initial benefit during the follow-up period, endoscopic manometry and endoscopic sphincterotomy were performed. All patients then received further prospective clinical follow-up examinations. RESULTS: With the exception of one patient with mild pancreatitis (4.5%), no side effects were observed after endoscopic BTX injection. Six weeks after BTX injection, 12 SOD patients (55%) were symptom-free, but ten patients (45%) were not. However five of these ten SOD patients who did not experience symptomatic benefit from BTX injection had normal basal sphincter of Oddi pressures (< 40 mmHg) at this time, and none of these five patients was free of complaints after subsequent endoscopic sphincterotomy. Two of the remaining five patients with sustained sphincter hypertension after BTX injection benefitted from endoscopic sphincterotomy. Eleven of the 12 SOD patients who had initially responded to BTX injection developed recurrent symptoms after a median period of six months. Manometry revealed sphincter hypertension in all 11 cases, and all patients became free of complaints again after endoscopic sphincterotomy during a median follow-up of a further 15 months. Overall, 11 of the 12 patients who responded to BTX injection, versus two of the ten patients who did not gain pain relief after BTX injection, later benefitted from endoscopic sphincterotomy (p < 0.01). CONCLUSIONS: Endoscopic injection of botulinum toxin into the papilla of Vater is a safe procedure and provides short-term relief of symptoms in half of patients with type III SOD. Our results also indicate that the clinical response to BTX injection can predict whether SOD patients will gain long-term benefit from endoscopic sphincterotomy. PMID- 9865561 TI - Microbiological examinations and in-vitro testing of different antibiotics in therapeutic endoscopy of the biliary system. AB - BACKGROUND AND STUDY AIMS: Prior to endoscopic therapeutic procedures, no antibiotic prophylaxis is administered routinely. Because of the reported incidence of infectious complications, which may reach up to 10%, a prospective study was undertaken to investigate the effects of a prophylactic dose of cefuroxime on the incidence of bacteremia and clinical signs of infection, but no significant effects could be demonstrated. In addition to this published work, blood and bile cultures obtained in this trial were also investigated, and the in vitro susceptibility to several antibiotics was tested in order to recommend the appropriate substances. PATIENTS AND METHODS: Ninety-nine consecutive patients (51 men, 48 women; mean age 61.4 +/- 17 years) with biliary obstruction who underwent an endoscopic retrograde cholangiopancreatography (ERCP) or percutaneous transhepatic cholangiography with drainage (PTCD) were included. Sequential blood cultures were taken before and up to 60 minutes after the endoscopic intervention. Bile cultures were obtained in 56 patients with biliary drainage. Aerobic and anaerobic cultures were prepared from all obtained specimens and the isolated organisms were identified. In the case of positive cultures, an in-vitro resistance test for 15 different antibiotics was performed. RESULTS: The incidence of bacteremia was 11.1% (n = 11), and 16 bacteria were isolated. Twelve different microorganisms were detected, with Escherichia coli found in four cases. From 41 positive out of 56 prepared bile cultures (73.2%), 91 isolates were found with 25 different species. A single agent was detected in eight cases (19.5%), while a mixed growth, with pathogens ranging from two to six species, was found in 33 cases (80.5%). The seven most frequently isolated germs were E. coli and Enterococcus (each n = 19), Klebsiella (n = 10), Streptococcus viridans (n = 9), Staphylococcus epidermidis (n = 5), Morganella morganii (n = 4), and Bacteroides fragilis (n = 3), representing 76% of all agents. Examination for fungal infection revealed positive cultures of Candida albicans in 16.1% of bile cultures (nine of 56). Interestingly, the use of proton-pump inhibitors (PPIs), with a consequent rise in the gastric pH value, led to an increase in the rate of bacteremia to 26.2% (five of 19) compared to the other patients not on PPIs (n = 80), who developed bacteremia in only six cases (7.5%; p = 0.02). In vitro testing of different antibiotics was carried out in 73 isolates. Imipenem showed the best antimicrobial activity (98.4%), followed by trimethoprim and sulfamethoxazole (90%), amoxicillin plus clavulanic acid (87.3%), vancomycin (82.4%), and ofloxacin (76.9%). CONCLUSIONS: Escherichia coli was found to be the pathogen most frequently detected in blood and bile following endoscopic interventions in the biliary tract. Enterococci, Klebsiella and Streptococcus viridans were found in bile cultures with an incidence exceeding 10%. In view of the in-vitro test results, possible side effects, and contraindications, amoxicillin plus beta-lactamase inhibitors or quinolones are considered to be suitable antibiotics for the prophylaxis of biliary infections. PMID- 9865562 TI - Experimental assessment of endoscopic mucosectomy with a cap-fitted panendoscope. AB - BACKGROUND AND STUDY AIMS: The use of a cap-fitted panendoscope is one method of carrying out endoscopic mucosectomy in the esophagus, stomach, and large intestine. The purpose of this study was to determine the optimal volume of physiological saline for submucosal injection, the rate of mucosal extension after saline injection, the initial size of the resected mucosal specimen, and the most appropriate heights for the fitted caps used in the colon and in the rectum, respectively. METHODS: Endoscopic mucosectomies using cap-fitted panendoscopes were carried out on resected surgical specimens from ten patients with colorectal cancer. RESULTS: It was necessary to inject 12 ml of saline under the mucosa to prevent perforation. Submucosal saline injection extended the mucosa by 1.4 +/- 0.2 times. A cap with a height of 7 mm is suitable for performing mucosectomy in the colon safely, while both 7 mm and 10 mm caps can be used in the rectum. The initial size of the resected mucosal specimens obtained with both caps was 12-20 mm (mean 14 mm) in diameter, with no significant differences. As the sizes of resected mucosal specimens reported in the past have been obtained after submucosal saline injection, it appears that larger specimens can be resected with the cap-fitted panendoscope than with conventional methods. CONCLUSIONS: The conditions under which endoscopic mucosectomy using the cap fitted panendoscope can be performed safely in the colon and the rectum were suggested by this experimental study using resected specimens. A saline injection of 12 ml under the mucosa is necessary to prevent perforation. A cap with a height of 7 mm is the most suitable size for the colon, while both 7 mm and 10 mm caps can be used in the rectum. PMID- 9865563 TI - Biliary stenting with a prototype expandable Teflon endoprosthesis. AB - BACKGROUND AND STUDY AIMS: A new balloon-expandable Teflon endoprosthesis was developed for endoscopic placement in the biliary tree. After deployment to its maximum diameter the stent is hardened by exposure to UV light. We present our preliminary results with a prototype 18 Fr prosthesis. PATIENTS AND METHODS: Three patients with a distal common bile duct obstruction due to unresectable pancreatic head carcinoma were treated with this stent. Insertion of the stent was the only palliative procedure. Patients were followed up until stent dysfunction or death. RESULTS: Stent placement was successful in all patients and no procedural complications occurred. Distal migration occurred in one patient, clogging of the stent after 149 days in another. CONCLUSION: In patients with a malignant biliary stricture this newly developed expandable endoprosthesis is effective in relieving obstructive jaundice. The technique has potential advantages over placement of current expandable metal mesh stents. PMID- 9865564 TI - Mass screening programs in Japan: what can we learn in the West? PMID- 9865565 TI - Diagnosis and treatment of patients with severe hematochezia: a time for change. PMID- 9865566 TI - The perfect biliary stent: the search goes on. PMID- 9865567 TI - One-step endosonography-guided drainage of a pancreatic pseudocyst: a new technique of stent delivery through the echo endoscope. AB - We report here the first case of a one-step endosonography(EUS)-guided pseudocyst drainage. A prototype large channel curved array echo endoscope (Pentax FG-38 UX) and a prototype delivery system for placement of an endoprosthesis was used for the procedure. The delivery system (GIP MedicinTechnik GmbH/Medi-Globe Corporation) consists of a handle part with a piston, a metal ring sheath, a plastic catheter with a diathermy needle and a double pigtail endoprosthesis (8.5 Fr). When mounted on the endoscope the endoprosthesis can be advanced out of the distal end of the endoscope. The introduction of the stent as well as the stent release can be monitored entirely by ultrasound. The procedure was tested in a 76 year-old woman with a pseudocyst measuring 60 mm in diameter located in the tail of the pancreas. The procedure was well tolerated by the patient, and there were no procedural complications. The advantage of a large channel echo endoscope and our new prototype delivery system is that the endoprosthesis can be inserted in to a pancreatic cyst guided exclusively by EUS without exchange of endoscopes, catheters or guide wires. Further studies are warranted. PMID- 9865568 TI - Intralesional steroid injections for corrosive induced pyloric stenosis. AB - Gastric outlet obstruction is a well-recognized sequela of corrosive ingestion. Such patients are traditionally treated surgically. We have treated three such patients with intralesional steroid injections combined with through-the-scope balloon dilation. All three patients responded well and are asymptomatic after 12 months, 14 months and 3 years and 3 months of follow-up, respectively. Endoscopic balloon dilation combined with intralesional steroid may be an effective alternative to surgery in such patients. PMID- 9865569 TI - Guidelines of the French Society of Digestive Endoscopy (SFED): esophageal prosthesis. PMID- 9865570 TI - Endoscopic resection of large polypoid leiomyoma: case study. PMID- 9865571 TI - Pancreaticopleural fistula treated by transpapillary implantation of a plastic prosthesis. PMID- 9865572 TI - Role of endoscopic ultrasonography in guiding intralesional steroid injections in esophageal strictures. PMID- 9865573 TI - Orbital hemorrhage as a complication of gastroscopy. PMID- 9865574 TI - Pulmonary, cerebral and coronary emboli during bucrylate injection of bleeding fundic varices. PMID- 9865575 TI - Endoscopic removal of a toothbrush using biopsy forceps and thread. PMID- 9865576 TI - Endoscopic extraction of a covered esophageal Z-stent with the aid of Endoloops. PMID- 9865577 TI - "Cascade stomach": laparoscopic treatment. PMID- 9865578 TI - Inflammatory polyps: an unreported side effect of argon plasma coagulation. PMID- 9865579 TI - Mesenteric panniculitis spreading to the pancreas simulating pancreatic neoplasm. PMID- 9865580 TI - Removable self-expanding metal stents: a pilot study for treatment of achalasia of the esophagus. PMID- 9865581 TI - Internal hemorrhoids coexisting with chronic anal fissure: new nonsurgical modalities. PMID- 9865582 TI - Endoscopic metallic stenting and intraductal ultrasonography in a case of bile duct cancer. PMID- 9865583 TI - A signet-ring cell carcinoma of the stomach endoscopically recognized as a tiny discolored spot. PMID- 9865584 TI - Aspartic acid in the arcuate nucleus attenuates the depressive effects of naloxone on ventilation. AB - Ventilation, oxygen consumption, the ventilatory equivalent for oxygen, and ventilatory responses to hypoxia and to hypercapnia were evaluated in conscious male rats who received each of four treatments: (1) microinjection of artificial cerebrospinal fluid (aCSF) into the arcuate nucleus and subcutaneously saline (CS); (2) aspartic acid into the arcuate nucleus and saline subcutaneously (AS); (3) aCSF into the arcuate nucleus and naloxone subcutaneously (CN); and (4) aspartic acid into the arcuate nucleus and naloxone subcutaneously (AN). Rats treated with CN exhibited a depression of ventilation, ventilatory equivalent, ventilatory response to hypercapnia, and tidal volume response to hypoxia and to hypercapnia. AS had no effect on any parameters. Administration of both aspartic acid and naloxone attenuated all the effects of CN except the depression of minute ventilation in response to hypercapnia. Therefore the naloxone (a mu opioid receptor antagonist) induced a depression of ventilation that was attenuated by aspartic acid acting on N-methyl-D-aspartic acid receptors in the arcuate nucleus. PMID- 9865585 TI - Does the motor cortical control of the diaphragm 'bypass' the brain stem respiratory centres in man? AB - In humans, cortico-motor excitation of the diaphragm may act directly on the phrenic motor nucleus via the cortico-spinal tract 'bypassing' brain stem respiratory centres (RC); alternatively, or in addition, this control may be indirect via the RC and bulbo-spinal paths. To investigate this, we stimulated the motor cortex using transcranial magnetic stimulation (TMS) in six subjects at end-expiration (diaphragm relaxed) and during voluntary inspiration. The sizes of the evoked compound action potentials in the diaphragm and also, as a control, in the thumb were no different whether TMS was delivered during normocapnia or during hypocapnia (PET(CO2) = 25 mmHg) when, presumably, the respiratory 'oscillator' was silent. In a further six subjects, TMS was performed during relaxed spontaneous breathing at three different points in the respiratory cycle. No perturbations in respiratory pattern (either tidal volume or respiratory timing) were seen. Thus we have been unable to demonstrate that the cortico-motor excitation of the diaphragm acts via the brain stem RC. PMID- 9865586 TI - Alterations in respiratory neuronal activities in the medullary 'pre-Botzinger' region in hypocapnia. AB - 'Pre-inspiratory' neuronal activities in a rostral ventrolateral medullary 'pre Botzinger' complex have been hypothesized to generate eupnea. Respiratory modulated neuronal activities were recorded in this region in decerebrate, vagotomized, paralyzed, and ventilated cats, having bilateral carotid sinus nerve sections. As end-tidal partial pressures of carbon dioxide were reduced to hypocapnic levels, all neuronal activities which were tonic or expiratory inspiratory ('pre-inspiratory') either ceased or lost respiratory-modulation. Similarly, most expiratory and inspiratory expiratory activities did not maintain a phasic discharge. Half of the inspiratory neuronal activities did continue a phasic discharge, which commenced after phrenic activity or became independent of the phrenic rhythm. Results do not support a fundamental role of the 'pre Botzinger' complex in the neurogenesis of eupnea. Some neuronal activities can establish a phasic discharge in hypocapnia which is independent of the central respiratory rhythm. At normocapnia, this independent discharge is superseded and incorporated into the ponto-medullary respiratory neuronal circuit which generates eupnea. PMID- 9865588 TI - Quantitative analysis of gas losses and gains in the middle ear. AB - Quantitative analysis of the amount of gas entering and leaving the middle ear (ME) was performed in patients with central perforation or with ventilating tubes. Patients were divided into a 'balanced pressure group' (BP) where pressure remained close to atmospheric throughout the measuring period and an 'unbalanced pressure group' (UBP) where pressure decreased continuously throughout measurements. The rate of gas gain was 41.0 +/- 19.9 microl/h (mean +/- SD, n = 17) in the BP group, significantly higher than in the UBP group (21.4 +/- 9.6 microl/h, n = 8). The rate of gas loss was 34.8 +/- 17.7 and 36.1 +/- 15.9 microl/h in the BP and UBP groups, respectively. Deglutition frequency was 27 +/- 13.4 deglutition/h in the BP group, significantly higher than the deglutition frequency of the UBP group (16 +/- 8.9 deglutition/h). It seems that the amount of gas entering the ME per deglutition is rather stable and does not change significantly with ME pressure level. Negative ME pressure may result from decreased gas gain rate due to increased intervals between deglutitions and loss of interdependence between the rate of gas entering and leaving the ME. PMID- 9865587 TI - Respiratory work in elastase treated hamsters. AB - Biomechanical adaptations of the diaphragm in the hamster model of emphysema are similar to those observed in skeletal muscle with exercise training. The aim of this study was to evaluate whether the dynamic pressure-volume (PV) work of breathing in hamsters with elastase-induced emphysema may contribute to these adaptations. PV work in elastase treated animals was compared to healthy controls. The studies were performed in adult hamsters 14-16 months following intratracheal administration of elastase (elastase treated group, ET) or saline (control group, CTL). Airway and esophageal pressures and air flows were measured during spontaneous breathing in anesthetized, supine animals. Pulmonary work (WL) was computed from transpulmonary pressures and airflows. Functional residual capacity (FRC) and total lung capacity (TLC; defined as volume at 25 cmH2O) in ET were increased 2 and 1.8 times, respectively, compared with CTL. Averaged tidal volume (VT) and inspiratory flows were comparable between groups. Total work of breathing (WT) normalized per ml VT was not significantly affected with elastase treatment but the pulmonary elastance work (WE) was significantly less in ET animals than controls (0.88 +/- 0.61 g cm(-2) vs. 1.63 +/- 0.32). Pulmonary resistive work was not significantly different between ET and CTL animals. These results suggest that biomechanical adaptations of the diaphragm observed in ET hamsters are caused by mechanisms other than the changes in dynamic mechanical properties of the lung following elastase treatment. PMID- 9865589 TI - Effect of Fenoterol on PAF-induced lung edema in isolated and perfused rabbit lungs. AB - We have studied the effects of fenoterol on PAF-induced response in pulmonary circulation. We used 28 isolated and perfused rabbit lungs preparations: eight control preparations (CP), four vehicles preparations (VP), eight PAF preparations (PP) with two doses of PAF, one called low dose (LD = 0.5 microg/kg of weight) and the other high dose (HD = 1 microg/kg of weight) and eight Fenoterol preparations (FP) which we administered 0.05 mg of Fenoterol for 15 min, followed by a LD and HD of PAF. FP prevented elevation of pulmonary artery pressure (Ppa) as compared to PP, at LD of PAF: 12.615 (CI 95%: 8.57-20.885) versus 83.705 (CI 95%: 50.55-114.3) cm of water; and at HD of PAF: 19.38 (CI 95%: 11.235-28.94) versus 205.1 (CI 95%: 141.3-271) cm of water respectively. FP prevented the increase in fluid filtration rate (FFR) observed in PP at both doses of PAF LD: 0.765 (CI 95%: 0.07-3.385) versus 0.01 (CI 95%: -0.05-0.005) g/min; HD: 5.515 (CI 95%: 2.425-8.865) versus 0.03 (CI 95%: 0-0.33) g/min. Our results suggest that PAF has a vasoconstrictor effect that produces lung edema and this effect is inhibited by fenoterol. PMID- 9865590 TI - Segmental vasodilatory effectiveness of inhaled NO in lungs from chronically hypoxic rats. AB - Inhaled nitric oxide (iNO) is being used to treat pulmonary hypertension in a variety of chronic lung diseases associated with pulmonary vascular remodeling. We hypothesized that chronic hypoxia (CH)-induced vascular remodeling decreases the vasodilatory effectiveness of iNO due to a thickened diffusional barrier. We therefore examined segmental vasodilatory responses to iNO in U-46619-constricted lungs isolated from control and CH (4 weeks at 0.5 atm) rats using double occlusion technique. We further measured lung fluid flux and vascular wall thickness in lungs from each group to provide an index of vascular permeability and vascular remodeling, respectively. CH was associated with decreased venous, but not arterial, responsiveness to iNO in saline-perfused lungs. In addition, the presence of red blood cells (RBC) within the perfusate greatly reduced venodilation in both groups of lungs, indicating that venous responsiveness to iNO in saline-perfused lungs is largely dependent upon transport of NO from an upstream site. In contrast, RBC had no effect on arterial dilation in control lungs, but attenuated arterial dilation to iNO in lungs from CH rats. Finally, fluid flux and arterial wall thickness were greater in lungs from CH rats. We conclude that arterial remodeling associated with CH may limit venous dilation to iNO. Furthermore, the decreased arterial responsiveness to iNO following CH is consistent with extravasation of hemoglobin within the arterial vasculature. PMID- 9865592 TI - Relationship between CO2 drive and characteristics of apneas in obstructive and central sleep apnea. AB - Previously we showed that CO2 drive is increased in patients with obstructive sleep apnea (OSA). In the present study we would like to evaluate a possible relationship between CO2 drive and characteristics of apneas in obstructive and central sleep apnea (CSA). We compared the hypercapnic ventilatory response (HCVR) between patients with OSA and CSA. HCVR was correlated with total event time and mean event duration in both groups. 17 normocapnic patients in each group and 14 controls were studied. The apnea patients were matched for apnea hypopnea index, age, sex, and BMI. SHCVR (slope) tended to be higher in apnea patients than in controls without statistical significancy: controls 1.65 (0.16), CSA 2.17 (0.22), OSA 2.55 (0.35) (l/min per mmHg) (P = 0.13). A significant correlation was found between HCVR and event time in CSA (0.52, P = 0.04) and between HCVR and apnea mean duration in OSA (r = 0.52, P = 0.04). We conclude that while CO2 drive was not statistically increased in both apnea types, small changes can contribute to breathing instability and may increase the total event time (in CSA) but may also shorten the apnea duration (in OSA). PMID- 9865591 TI - Effects of modulators of the production and degradation of hydrogen peroxide on erythropoietin synthesis. AB - Erythropoietin (Epo) synthesis is suppressed in normoxia and stimulated in hypoxia. To test the hypothesis that the cellular H2O2 level is important in the control of Epo synthesis, we have studied effects of modulators of H2O2 generation and degradation on Epo production in human hepatic cell cultures (hepatoma lines HepG2 and Hep3B). In addition, we measured the activities of antioxidant enzymes (catalase, superoxide dismutase, glutathione peroxidase) in cultures following hypoxia exposure or H2O2 treatment. The results show that the formation of immunoreactive Epo was stimulated in normoxic cultures by treatment with exogenous catalase thus mimicking the effect of hypoxia (24 h incubation periods). Epo production was also stimulated when scavengers of reactive O2 species (tetramethylthiourea, dihydrorhodamine) were added to the cells. On the other hand, stimulators of H2O2 generation (xanthine oxidase, glucose oxidase, NADH, NADPH) lowered Epo production in hypoxic cultures. Hypoxia exposure decreased superoxide dismutase activity and H2O2 treatment reduced catalase activity thus influencing the endogenous antioxidant defense system. These findings support the concept that reactive O2 species, primarily H2O2, act as messengers in the O2-dependent control of the hepatic production of Epo. Changes in the cellular activities of antioxidant enzymes appear to play only a minor role in this process. PMID- 9865593 TI - Exhaled nitric oxide in preterm infants. AB - Nitric oxide (NO) is detectable in the exhaled gas of adults during spontaneous respiration and, according to current knowledge, mainly originates from the paranasal sinuses. We studied total NO excretion rates by chemiluminescence in preterm infants whose paranasal sinuses are known to be only partially pneumatized. NO excretion was 7.15 +/- 1.13 nl/min (mean +/- SD, range 6.33-9.36 nl/min) measured from spontaneously exhaled nasal gas (n = 6) and 0.3 +/- 0.05 nl/min (range 0.26-0.36 nl/min) measured from the lower airways in intubated individuals (n = 3). These values are considerably lower than those reported for older children and adults. Body weight-related amounts of NO excretion, however, seem comparable between infants and adults. PMID- 9865594 TI - Imuthiol inhibits the etoposide-induced apoptosis in HL-60 cells. AB - Imuthiol (sodium-diethyl-dithiocarbamate, DTC) has been proven to possess immunomodulatory properties, this effect, however, varies in different model systems. This study was performed to assess the possible effect of imuthiol on etoposide-induced apoptosis in the human promyelocytic leukemia cell line, HL-60. In vitro pre-treatment with imuthiol at concentrations between 50 and 100 microM significantly inhibited etoposide-induced apoptosis of HL-60 cells, the inhibition appeared to be dose dependent. This effect may be associated with the immunomodulatory activity of imuthiol. PMID- 9865595 TI - Cytokine patterns in seropositive and seronegative patients with Echinococcus granulosus infection. AB - To investigate the involvement of immunological variables, including the Th1/Th2 balance, in the mechanisms underlying seronegativity in cystic echinococcosis (CE), Th1 and Th2 cytokines were evaluated in ELISA determining IFN-gamma, IL-4, 5 and -10 production in peripheral blood mononuclear cell (PBMC) cultures from 16 patients (six seronegative and ten seropositive) with CE and from six healthy donors. Patients who had undetectable antibody levels in their sera invariably produced no IL-5 and scarce IL-4 and -10 (Th2 response); both groups of patients produced similar amounts of IFN-gamma; (Th1 response). The analysis of host clinical features and cyst characteristics shows that these variables do not explain the lack of detectable serum antibody levels. These findings suggest that seronegativity arises because unknown host or parasite factors, or both, cause inadequate Th2 cell activation thus reducing production of Th2 cytokines, including IL-5, the cytokine primarily implicated in immunoglobulin expression in CE. PMID- 9865596 TI - Effect of pre-existing carrier immunity on the efficacy of synthetic influenza vaccine. AB - In our previous studies on the development of synthetic peptide-based vaccines, we have evaluated flagellin from a Salmonella typhi vaccine strain as a carrier molecule for synthetic peptides derived from the influenza virus. The results indicated that the use of recombinant flagella, expressing defined influenza epitopes, is adequate for induction of protection against a challenge infection. It is of importance to show that previous exposure to the carrier does not induce suppression of the response to such vaccine. In the present study we demonstrate that the protective effect is not impaired by pre-immunization with either the carrier flagellin molecule alone or with the intact salmonella. The immune response was manifested both by the level of antibodies produced and by a proliferative cellular response, as well as by an efficient protection of the mice from a sub-lethal challenge infection. Since prior exposure to the carrier did not result in immune suppression, we conclude that Salmonella flagellin is a suitable carrier for synthetic peptide based vaccines. PMID- 9865597 TI - Ca2+-ATPase inhibitor induces IL-4 and MCP-1 production in RBL-2H3 cells. AB - The effects of a Ca2(+)-ATPase inhibitor, cyclopiazonic acid (CPA), and two hydroquinone-antioxidants, 2,5-di-(tert-butyl)-1,4-hydroquinone (DTBHQ) and 2,5 di-(tert-amyl)-1,4-hydroquinone (DTAHQ) on the release of IL-4 and MCP-1 from RBL 2H3 cells were investigated. CPA, DTBHQ and DTAHQ, all of which induce intracellular free Ca2+ concentration ([Ca2+]i) increase, induced IL-4 and MCP-1 release in a dose-dependent manner. The release of TNF-alpha required both a Ca2(+)-ATPase inhibitor and 12-O-tetradecanoylphorbol-13-acetate (TPA). However, the Ca2(+)-ATPase inhibitors induced IL-4 and MCP-1 production without TPA. The release of IL-4 and MCP-1 reached a maximum at 9 and 6 h, respectively. IL-4 and MCP-I release was inhibited by treatment with the immunosuppressant FK-506 and actinomycin D. Therefore, in our system IL-4 and MCP-1 release involves Ca2(+) dependent and FK-506-sensitive signaling pathways. This is the first report about Th-2 type cytokine and chemokine production in RBL-2H3 cells. PMID- 9865598 TI - Identification of a soluble interleukin-8 inhibitor in the supernatant of polymorphonuclear leukocytes. AB - Interleukin-8 (IL-8) plays a crucial role in the pathogenesis of inflammatory and hyperproliferative diseases in various organs. The purpose of the present investigation was to establish whether there is any naturally occurring inhibitor of IL-8. Here we demonstrate that an IL-8 inhibitor (IL-8INH) is present in the supernatant of polymorphonuclear (PMN) leukocytes. The release of IL-8INH could be increased by stimulating the PMN leukocytes by concanavalin A. IL-81NH blocks the IL-8-induced chemotaxis and Candida albicans killing activity of PMN leukocytes and epidermal cells in vitro, and IL-8-induced neutrophil infiltration in the mouse ear in vivo. The mechanism of action of IL-8INH involves blocking of 125I-IL-8 binding to the IL-8 receptor. Binding of 125I-IL-8 to neutrophils could not be displaced by the IL-8INH, however, preincubation of 125I-IL-8 with IL-8INH increased binding inhibition, suggesting an interaction between IL-8 and the inhibitor. Crosslinking of 125I-IL-8 to IL-8INH shows that IL-8INH binds specifically to 125I-IL-8, and the IL-8INH protein has an apparent molecular weight of 52 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The partial purification of the IL-8INH on DEAE-Sephadex anion-exchange chromatography column also suggests a 50-60-kDa inhibitor protein which blocks IL 8-induced effects on neutrophils by binding to IL-8. PMID- 9865599 TI - Expression of a Fas-like proapoptotic molecule on the lymphocytes of Xenopus laevis. AB - Ligation of the externally expressed Fas (APO1/CD95) molecule will initiate programmed cell death (apoptosis), in many mammalian developing and adult cells. Fas-induced apoptosis has not been demonstrated with the cells of any non mammalian vertebrate. We immunostained suspensions of splenocytes from adult Xenopus laevis, the South African clawed toad, with a polyclonal rabbit anti human Fas antibody raised against the amino acid residues 321-335 of human Fas. The binding was specific, as it was dramatically reduced by preincubation of the antibody with the Fas peptide used to make it, but not with a Fas-ligand (FasL) peptide. The binding was enhanced after in vitro exposure of the splenocytes to phytahemagglutinin (PHA), a T cell mitogen and apoptogen in this species. Sections of developing Xenopus larval tissue were also immunostained with the polyclonal rabbit anti-human Fas antibody. Consistent binding of thymocytes and splenocytes was not observed until early metamorphosis in these immunological sites. A monoclonal mouse anti-human Fas antibody, previously used to stimulate apoptosis in mammalian cells, induced significant levels of apoptosis in adult Xenopus splenocytes and additionally, bound specifically to a splenocyte extract, as assayed by ELISA. Thus, a molecule on Xenopus splenocytes shares both structural and functional homologies with human Fas, indicating the evolutionary conservation within vertebrates of this means of initiating apoptosis. PMID- 9865600 TI - Preferential tissue localization of bovine gamma delta T cell subsets defined by anti-T cell receptor for antigen antibodies. AB - Clonal and oligoclonal populations of gammadelta T cells, with respect to the expression of T cell receptors for antigen (Tcr), have been shown to localize in normal and inflamed tissues. The mechanisms responsible for the tissue-selective accumulation of these subsets are still not known. gammadelta T cells are the predominant T cell subset in newborn calves, making this animal a useful model to study these cells. However, molecular markers defining tissue-specific bovine ydelta T cell subsets are only now being developed. In this report, we describe three new anti-bovine gammadelta Tcr mAbs: GD3.8, GD197 and GD3.1, which provide useful tools to study these cells. GD3.8 recognized virtually all gammadelta T cells in the blood; whereas GD3.1 and GD197 recognized mutually exclusive as well as overlapping subsets. Using these three mAbs, four separate subsets of gammadelta T cells were defined: subset 1 (GD3.8+, GD3.1+, GD197-); subset 2 (GD3.8+, GD3.1-, GD197+); subset 3 (GD3.8+, GD3.1+, GD197+); and subset 4 (GD3.8+, GD3.1-, GD197-). Subset 4 constituted a minor population in the blood; however, it predominated in the spleen and, in some cases, represented a 300% increase over blood levels. The percentage of GD3.1-positive gammadelta T cells was found to be increased in experimentally inflamed lymph nodes, suggesting that subset 1 cells may be preferentially retained in or recruited to sites of inflammation. Some subset 4 cells also exhibited a decreased ability to respond to PHA. These studies demonstrate that bovine gammadelta T cell, Tcr-defined subsets, exhibit unique accumulation and activation characteristics that may provide clues to their function and regulation. PMID- 9865601 TI - Chloroquine inhibits HIV-1 replication in human peripheral blood lymphocytes. AB - Chloroquine has inhibitory effects on human immunodeficiency deficiency virus (HIV)-1 replication in human lymphocytes with an ED50 of 15 microM, which approximates the chloroquine plasma concentration generated in the acute treatment of malaria. Chloroquine has a parallel inhibitory effect on DNA replication in human lymphocytes, although the inhibitory effect on DNA replication is not as pronounced as is the drug inhibition of HIV-1 replication in the cells. PMID- 9865602 TI - Elevation of serum IgE level and peripheral eosinophil count during T lymphocyte directed gene therapy for ADA deficiency: implication of Tc2-like cells after gene transduction procedure. AB - We have successfully carried out T-cell-directed gene therapy for a boy with severe combined immunodeficiency due to adenosine deaminase deficiency (ADA SCID) and unexpectedly found an elevation of serum IgE level and peripheral eosinophil count during the course. More than 90% of transduced cells cultured for 7-11 days before infusion into the patient were positive for CD8 and expressed Th2-type cytokine genes such as IL-4, IL-5 and IL-13. Furthermore, CD4(+) T-depleted PBMC (peripheral blood mononuclear cells) from the patient synthesized IgE in vitro by stimulation with IL-4. Collectively, these results suggested that Tc2-like cells in the transduced cells have distinct immunological functions to help IgE synthesis and activate eosinophils. PMID- 9865603 TI - Rapid acquisition of beta-sheet structure in the prion protein prior to multimer formation. AB - The N-terminally truncated form of the prion protein, PrP 27-30, and the corresponding recombinant protein, rPrP, were solubilized in 0.2% SDS, and the transitions induced by changing the conditions from 0.2% SDS to physiological conditions, i.e. removing SDS, were characterized with respect to solubility, resistance to proteolysis, secondary structure and multimerization. Circular dichroism, electron microscopy and fluorescence correlation spectroscopy were used to study the structural transitions of PrP. Within one minute the alpha helical structure of PrP was transformed into one that was enriched in beta sheets and consisted mainly of dimers. Larger oligomers were found after 20 minutes and larger multimers exhibiting resistance to proteolysis were found after several hours. It was concluded that the monomeric alpha-helical conformation was stable in SDS or when attached to the membrane; however, the state of lowest free energy in aqueous solution at neutral pH seems to be the multimeric, beta-sheet enriched conformation. PMID- 9865604 TI - Cryoinactivation and conformational drift of glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle. AB - The cryoinactivation of glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle (GAPDH-rabbit) was studied. It was found that the inactivation of GAPDH rabbit at 0 degrees C was much faster than that of GAPDH from yeasts, and showed obvious time and concentration dependence. The spectral properties, enzyme activity and behavior under pressure, of GAPDH-rabbit treated either by cryoinactivation, or pressure-induced dissociation and reassociation, were very similar. These results provided evidence to support the idea that cryoinactivation of oligomeric proteins, might take place through a cycle of dissociation-reassociation accompanied with the so-called conformational drift postulated by King and Weber (1986). PMID- 9865605 TI - A parallel association between differentiation and induction of galectin-1, and inhibition of galectin-3 by retinoic acid in mouse embryonal carcinoma F9 cells. AB - Soluble endogenous lactoside-binding lectins, galectins, have been implicated in cell adhesion, growth, differentiation, neoplastic transformation, and metastasis. Two major classes of these lectins, galectin-1 and galectin-3, are developmentally regulated. To explore the mechanisms by which the expression of the galectins is regulated and to examine their association with the differentiation processes induced by all-trans retinoic acid (RA), dibutyryl cyclic AMP (Bt2cAMP) and their combination, we used the murine embryonal carcinoma (EC) cell line F9 and its RA-resistant mutant, RA-3-10. RA induced endodermal differentiation and a concurrent induction of galectin-1 and its complementary glycoconjugates (laminin and lysosomal-associated membrane protein, LAMP) in the F9 wild-type (wt) line, but failed to induce differentiation and had no effects on or even reduced the expression of galectin-1, laminin, and LAMP in the RA-3-10 line. On the other hand, RA inhibited expression of galectin-3 in the wild-type line but had no effect on the RA-3-10 line. The galectin-1 gene is at least partially regulated at the transcriptional level. These results demonstrate a parallel association between differentiation and induction of galectin-1, and inhibition of galectin-3 in F9 cells by RA. The study suggests that a regulated expression of galectins and their complementary glycoconjugates is involved in the differentiation pathway induced by RA in F9 cells. PMID- 9865606 TI - Transcription of the rat p53 gene is mediated by factor binding to two recognition motifs of NF1-like protein. AB - In this study we analyzed the ratp53 promoter by electrophoretic mobility shift assay (EMSA) and DNase I footprinting analysis. As a result we identified two protein binding elements (element 1: -296 to -312, element 2: -195 to -219) with sequence homology to each other. The two identified elements bind to the same kind of protein. To identify the protein binding to these elements, competition assays were carried out with double stranded oligonucleotides containing NF1, YY1, and CRE consensus motifs. Only the NF1 consensus motif competed with element 1 and 2. Element 2 is conserved between the rat, human, and mouse p53 promoters, and has an NF1 consensus motif. However, the sequences of element 1 are comparatively variable between the species. Only the element 1 region of the rat p53 promoter has partial homology to the NF1 consensus motif. This suggests that the element 1 is specific for the rat p53 gene. The molecular mass of the binding protein, determined by Southwestern blotting analysis, was 40 kDa, which is different from that of NF1. In EMSA with an anti-NF1 antibody, DNA-protein complexes were neither supershifted nor decreased. The 40 kDa protein was also detected in rat spleen and lung, but not in kidney. The binding protein was purified by sequence-specific DNA affinity chromatography and it was confirmed that the purified protein binds to the two regions. It was also proved that the identified two elements are required for basal level transcription of the rat p53 gene by in vitro transcription assay. PMID- 9865608 TI - The glutamine synthetase from the hyperthermoacidophilic crenarcheon Sulfolobus acidocaldarius: isolation, characterization and sequencing of the gene. AB - The glutamine synthetase (EC 6.3.1.2) from the hyperthermoacidophilic crenarcheon Sulfolobus acidocaldarius (DSM 639) was purified to homogeneity, characterized and the glnA gene isolated and sequenced. The amount of enzyme present in the cytosolic fraction from Sulfolobus cells showed a strong variation depending on the carbon and nitrogen sources in the growth medium. The enzyme was found to be a dodecameric protein composed of identical subunits of 52 kDa. It was stable at 78 degrees C in the presence of Mn2+ ions. The catalytic activity was regulated solely by feed-back inhibition through L-alanine and glycine and not by adenylylation. No evidence for the presence of isoenzymes was found. Sequence comparison showed that the Sulfolobus protein is most closely related to the glutamine synthetases of the I-beta type despite its regulatory properties and the finding that the known euryarcheal glutamine synthetase sequences belong to the I-alpha subgroup of these enzymes. Our phylogenetic analysis suggests that the gene duplication leading to the development of the I-alpha and I-beta enzymes preceded the separation of the archea and the bacteria. PMID- 9865607 TI - Inhibition of hepatocytic autophagy by adenosine, adenosine analogs and AMP. AB - Autophagy, measured in isolated rat hepatocytes as the sequestration of electroinjected [3H]raffinose, was moderately (17%) inhibited by adenosine (0.4 mM) alone, but more strongly (85%) in the presence of the adenosine deaminase inhibitor, 2'-deoxycoformycin (50 microM), suggesting that metabolic deamination of adenosine limited its inhibitory effectiveness. The adenosine analogs, 6 methylmercaptopurine riboside and N6,N6-dimethyladenosine, inhibited autophagy by 89% and 99%, respectively, at 0.5 mM, probably reflecting the adenosine deaminase resistance of their 6-substitutions. 5-Iodotubercidin (10 microM), an adenosine kinase inhibitor, blocked the conversion of adenosine to AMP and largely abolished the inhibitory effects of both adenosine and its analogs, indicating that AMP/nucleotide formation was required for inhibition of autophagy. Inhibition by adenosine of autophagic protein degradation, measured as the release of [14C]valine from prelabelled protein, was similarly potentiated by deoxycoformycin and prevented by iodotubercidin. Inhibition of autophagy by added AMP, ADP or ATP (0.3-1 mM) was, likewise, potentiated by deoxycoformycin and prevented by iodotubercidin, suggesting dephosphorylation to adenosine and intracellular re-phosphorylation to AMP. Suppression of autophagy by AMP may be regarded as a feedback inhibition of autophagic RNA degradation, or as an aspect of the general down-regulation of energy-requiring processes that occurs under conditions of ATP depletion, when AMP levels are high. PMID- 9865609 TI - RNA splicing of bacterial genes in eukaryotes. AB - The presence of intervening sequences or introns in eukaryotic genes has been known for more than 20 years, and the mechanisms underlying RNA splicing have been studied in depth both genetically and biochemically. In recent years, however, an increasing number of bacterial genes have been introduced into higher eukaryotes as important tools for genetic studies. Their gene products are frequently used as an indirect measure for cell type-specific promoter activity, as, for example, in the case of chloramphenicol acetyl transferase (CAT assay) or beta-galactosidase. Here we show that RNA splicing of two prokaryotic genes encoding site-specific DNA recombinases occurs in eukaryotic cells. In one case, splicing is only observed after treatment of cells with the cytokine alpha interferon. We further demonstrate that mutating an intragenic donor splice site in a bacterial gene apparently activates a second, alternative splicing pathway. In conjunction with previous reports, our findings should also be regarded as a warning that splicing of bacterial genes in higher eukaryotes is a more common phenomenon than presently recognized, which may be difficult to overcome and may cause problems in the interpretation of experimental results. PMID- 9865610 TI - Characterization of the chicken and rat DCoH gene domains using an improved ligation-mediated PCR method. AB - The bifunctional protein DCoH (dimerization cofactor of HNF-1) is a transcriptional coactivator of homeodomain proteins and a cytoplasmic enzyme. Here we report on the isolation, nucleotide sequence and genomic structure of the chicken and rat DCoH genes. They have an unusual first exon coding for a single amino acid. The promoters are located within CpG-islands and lack TATA and CAAT consensus sequences. Analysis of the number of CpG dinucleotides in the promoter indicates little changes since man and chicken were separated in evolution whereas a 67% reduction was observed in the rat lineage. The DNA sequences were obtained entirely without cloning. For walking into unknown genomic regions we describe a modification of the ligation-mediated polymerase chain reaction (LM PCR) that can replace conventional screening and cloning methods and greatly expedite the isolation of genomic DNA. PMID- 9865611 TI - Isolation and characterization of rabbit plasma alpha-1-antiproteinase E. AB - Alpha-1-antiproteinase E, the fourth isoform of rabbit alpha-1-antiproteinase (alpha-1-antitrypsin) having a glutamic acid at the reactive center, has been purified from the plasma by sequential chromatography on hydroxyapatite and anion exchange columns. The E form of alpha-1-antiproteinase formed a complex with trypsin, chymotrypsin, elastase, plasmin and pancreatic kallikrein as judged by SDS-PAGE. The E form inhibited elastase in a stoichiometric manner and chymotrypsin moderately, but the inhibition of trypsin was gradual. The F form inhibited trypsin most effectively followed by chymotrypsin and elastase. N chlorosuccinimide reduced the elastase inhibitory activity of the E form, while the F form was more effectively inactivated by the oxidant. PMID- 9865612 TI - Cystatin and cystatin-derived peptides have antibacterial activity against the pathogen Porphyromonas gingivalis. AB - We investigated whether cystatins and cystatin-derived peptides, encompassing sequences of secondary structures of cystatin S and papain binding domains of cystatin C, display antimicrobial properties. Of the different microorganisms tested, only the growth of P. gingivalis was inhibited by chicken cystatin and cystatin C. Cystatin S, cystatin S:1-14, cystatin S:61-73 and cystatin S:108-121 also inhibited its growth, whereas cystatin S:21-38, cystatin S:39-55, cystatin S:81-95, cystatin S:94-109, and cystatin C: 9-12/55-60/106-107 did not. No inhibition of the cysteine proteinase activity of P. gingivalis was observed for all cystatin-derived peptides. On the other hand, leupeptin and antipain inhibited P. gingivalis proteinase activity, but had no effect on the growth. These data suggest that cystatins contain antibacterial sequences active against P. gingivalis and that the growth inhibition does not depend on the inhibition of P. gingivalis cysteine proteinases. PMID- 9865613 TI - Inactivation of intracellular and non-enveloped viruses by a non-ionic naphthalene endoperoxide. AB - Singlet oxygen (1O2, 1delta(g)) selectively oxidizes many biological targets, some of which, such as viruses, are located intracellularly under in vivo conditions. Considering the short lifetime of 1O2 in aqueous media, it is essential to generate this species in close proximity to the targets. Therefore, a water-soluble and non-ionic carrier of 1O2, DHPNO2, has been designed to convey 1O2 through lipid membranes. In contrast to the known anionic carrier NDPO2, which inactivates only extracellular enveloped viruses, the new compound exhibits virucidal activity on all types of viruses, enveloped (HIV) and non-enveloped (Poliovirus), extracellular and intracellular. HIV inactivation can also be achieved in the presence of red blood cells, suggesting the possible use of DHPNO2 in the decontamination of cellular blood products. PMID- 9865614 TI - Analysis of structure and microtubule assembly activity of the Drosophila 205K MAP. AB - Phosphorylation, dimerization and binding to calmodulin have been reported to influence the microtubule assembly capacities of MAPs (microtubule-associated proteins). Here we report that the Drosophila 205K MAP is a phosphoprotein in vivo and can be phosphorylated by cdc2/p34 in vitro. Bacterially produced 205K MAP is competent of microtubule assembly and microtubule bundling and binds to immobilized calmodulin in a Ca2+-dependent way. EM rotary shadowing analyses suggest that 205K MAP consists of an amino-terminal flexible extended region and a carboxy-terminal globular domain. This carboxy-terminal region harbors the microtubule binding site and sequences required for dimerization, as confirmed by in vitro crosslinking experiments of truncated proteins. PMID- 9865615 TI - Leech antihemostatic proteins share the T-knot scaffold, a disulfide-reinforced structural motif. AB - The occurrence of similar topologies among unrelated proteins is an emerging theme in structural biology. Here we report that the T-knot scaffold, a disulfide reinforced structural motif shared by knottins and EGF-like proteins, is also present in leech antihemostatic proteins. Our finding emphasizes the versatile nature of this small structural motif, representing a compact structural unit suitable for the diverse biological functions performed by knottins, EGF-like proteins and leech antihemostatic proteins. PMID- 9865616 TI - Cloning and characterization of the gene encoding a new DNA methyltransferase from Neisseria gonorrhoeae. AB - A HindIII fragment of N. gonorrhoeae MS11 DNA coding for DNA methyltransferase (MTase) activity was cloned and expressed in E. coli AP1-200-9 cells. The sequence of 4681 bp was determined, and its analysis revealed two open reading frames (ORFs) sharing some similarity with known DNA MTases. ORF1 encodes an active N4mC MTase (M.NgoMV). The enzyme modifies only one strand of double stranded DNA and preferentially recognises the sequence GCCHR although it is able to methylate other sites. The exact recognition sequence cannot be precisely defined due to a relaxed specificity. The second ORF shows high homology to 5mC Mtases, but we were unable to demonstrate DNA methylating activity of its product either in vivo or in vitro. PMID- 9865618 TI - Effect of exposure to balanced salt solution upon the hardness of the crystalline lens. AB - BACKGROUND: The nucleus confers most of the hardness upon the lens, water content decreases towards the centre of the nucleus and a relative dehydration accompanies increased hardness in some cataractous lenses. It is a possibility that exposure and incubation of the inner layers of the nucleus to balanced salt solution (BSS) can result in the softening of the nucleus. PURPOSE: This study aimed to investigate the effect of BSS upon the lens hardness. METHODS: Nuclear colour element of cataract was graded biomicroscopically. Following extracapsular cataract surgery the lens nucleocortex was divided into two equal parts and each half was allocated randomly to incubation in BSS or air for 5 min before the hardness of each section was assessed by an automated guillotine. RESULTS: Following incubation with BSS the mean force necessary to bisect the lens was 0.50 N and in the control air group the mean force was 0.64 N. The lenses in the BSS-treated group were consistently softer than those in the control group, with a mean softening of 18.3% (P=0.001). The amount of softening was not related to the nuclear colour (P=0.6) or age (P=0.1). CONCLUSION: Softening of the lens through physical disruption has previously been reported. This study describes the phenomenon of nuclear softening following exposure to BSS, indicating that lens softening can occur through biochemical means. PMID- 9865617 TI - The effect of acetazolamide on passive and active transport of fluorescein across the blood-retina barrier in retinitis pigmentosa complicated by macular oedema. AB - BACKGROUND: The carbonic anhydrase inhibitor acetazolamide (AZM) reduces macular oedema in some patients with retinitis pigmentosa. To better understand the oedema-reducing effect of AZM, the effect of AZM on passive permeability and active transport of fluorescein across the blood-retina barrier was studied in patients with retinitis pigmentosa and varying degrees of macular oedema. METHOD: The selection of patients was based on an introductory examination including vitreous fluorometry for qualitative assessment of the vitreous. Macular oedema was graded by fluorescein angiographic leakage. The effect of AZM on the transport properties of the blood-retina barrier was determined by differential spectrofluorometry, in a randomised, double-masked, cross-over study, comprising 2 weeks' treatment with AZM (500 mg/day) and 2 weeks' treatment with placebo. The penetration ratio, defined as the ratio between vitreous concentration 3 mm in front of the retina and the plasma integral, was determined for fluorescein and its metabolite fluorescein glucuronide at 30-60 min and at 120 min after fluorescein injection. Passive permeability and unidirectional permeability in the direction vitreous to blood, due to outward active transport of fluorescein, were determined in those cases where the curves for vitreous concentration of fluorescein could be fitted to a mathematical model. Visual acuity was tested by use of ETDRS standard logarithmic charts. RESULTS: Twenty-two patients volunteered to participate in the study. Signs of significant vitreous detachment/liquefaction caused the exclusion of ten patients after the introductory examination. Nine patients with approximately intact vitreous and varying degrees of oedema completed the cross-over study. AZM treatment was related to a decrease in the penetration ratio of 21% for fluorescein (P=0.01) and of 22% for fluorescein glucuronide (P=0.004). Passive permeability and unidirectional permeability were determined in seven patients. AZM caused a decrease of 27% in the passive permeability of fluorescein (from 1.1 x 10(1) nm/s, P=0.031), and a 95% increase in unidirectional permeability of fluorescein (from 1.2 x 10(2) nm/s, P=0.047). AZM led to a reduction in the grade of macular oedema as determined by fluorescein angiography in three out of seven patients. Only small improvements (< or =5 letters) in visual acuity were noted. CONCLUSION: The present study indicates that the oedema-reducing effect of AZM is due to decreased leakage and stimulated active transport across the blood-retina barrier. PMID- 9865619 TI - Analysis of tear proteins by one- and two-dimensional thin-layer iosoelectric focusing, sodium dodecyl sulfate electrophoresis and lectin blotting. Detection of a new component: cystatin C. AB - BACKGROUND: Isoelectric focusing (IEF) of tear proteins has not yet been carried out in a satisfactory way. Two-dimensional (2D) electrophoresis, especially in the combination of IEF with SDS, is able to differentiate between proteins in detail. The purpose of this study was therefore to analyze tear proteins by 1D IEF alone and in combination with a 2D pattern, and by IEF followed by lectin staining. METHODS: Ampholines, covering a broad range from pH 3 to pH 10, were applied. After IEF, semi-dry blotting and incubation with a group II lectin and two group V lectins was performed. RESULTS: Tear proteins could be separated into 31 single bands. Tear-specific pre-albumin (TSPA), lactoferrin, sIgA, IgG and lysozyme were found to be main components. Isoelectric points (IEPs, pls) of all proteins separated were determined by comparison with IEF standards. 2D patterns of IEF and SDS electrophoresis were obtained for the main subunit components of lactoferrin, sIgA, TSPA, and lysozyme. An additional new component of considerable concentration was focused at pI 8.6 with a subunit MW of 14 kDa. With s-WGA a component at an IEP of 5.2 was visualized, representing transferrin. With SNA, lactoferrin stained as a sharp main band at pI 5.1 with three additional weaker bands at IEPs from 4.8 to 4.9. At IEPs between 4.4 and 6.1, multiple components of sIgA were stained with MAA. The sugar specificity of transferrin at pI 5.2 was beta-GlcNAc. Lactoferrin showed glycation with NANA alpha-2-6-Gal or NANA-alpha-2-6-GalNAc, whereas the sugar specificity of sIgA was NANA-alpha-2-3-Gal. CONCLUSIONS: The investigative strategy applied here, including IEF alone, in combination with SDS-electrophoresis, and SDS electrophoresis followed by lectin staining proved to be a reproducible method for tear protein analysis of hitherto unexperienced capacity. Lectin-stained bands of native tear proteins are not uniformly glycated by one sugar residue, but show various sugar specificities. IgA as a whole molecule is specifically glycated with NANA-alpha-2-3-Gal. PMID- 9865621 TI - Retinal ganglion cell layer and visual function in a patient with optic disc drusen. AB - BACKGROUND: To correlate the retinal ganglion cell pattern to visual acuity and visual field data in a patient with bilateral optic disc drusen, a quantitative clinicopathological study was carried out. METHODS: Both retinae of a patient with optic drusen were whole-mounted. Retinal ganglion cell counts were made using a sampling scheme covering the whole retina and compared to the findings in 10 normal retinae. Relative ganglion cell reduction in the drusen retinae was correlated to clinical data. RESULTS: The total retinal ganglion cell count was reduced from 1244858+/-98736 in normal retinae to 305319 on the right and 527571 on the left eye with optic disc drusen. Large ganglion cells had a better chance of survival. Parafoveal ganglion cell loss was 57% for the right and 36% for the left eye, while visual acuity was 0.8 and 1.0 respectively. The mean light sensitivity loss increased from the centre (6.2 dB) to paracentral (9.9 dB), mid peripheral (13.7 dB) and outer peripheral (15.0 dB) retina, while ganglion cell losses were smallest in outer peripheral retina (21.9%), followed by central (53.0%), mid-peripheral (70.9%) and paracentral retina (87.7%). CONCLUSION: These data validate Frisen's theory on central retinal resolution and provide the structural basis for the clinical rule that low visual acuity should not be attributed to disc drusen. Automated light sense perimetry gives an inadequate picture of retinal damage caused by optic disc drusen. PMID- 9865620 TI - Substance P in proliferative vitreoretinopathy: the significance of aqueous humor levels for evolution of the disease. AB - PURPOSE: We detected aqueous humor levels of substance P in patients with various grades of proliferative vitreoretinopathy and with uncomplicated rhegmatogenous retinal detachment. To evaluate the significance of the concentration of substance P at the time of surgery for retinal detachment for subsequent development of proliferative vitreoretinopathy, the latter patients also underwent fundoscopic control examination. METHODS: Using a highly specific and sensitive radioimmunoassay, the content of substance P in fresh samples of aqueous humor obtained by paracentesis was determined both in cataract controls and in patients with uncomplicated rhegmatogenous retinal detachment and with various grades of proliferative vitreoretinopathy. Retinal detachment patients underwent fundoscopic control examination 6 months after surgical reattachment. RESULTS: The mean concentration of substance P in cataract controls was 40.3 (+22.4) fmol/mg protein, in the retinal detachment group 61.9 (+/-13.9) fmol/mg protein and in proliferative vitreoretinopathy 335.2 (+/-24.8) fmol/mg protein, but no correlation between levels of the peptide and various grades of the disease was observed. Already at surgery for retinal detachment three in four patients who developed proliferative vitreoretinopathy presented with levels of substance P in the range of the disease. CONCLUSION: The concentration of substance P in aqueous humor is significantly high in patients with proliferative vitreoretinopathy in whom surgery is indicated. Furthermore, elevation of the peptide in retinal detachment that originates most obviously from a neurogenic mechanism may indicate initiation of processes associated with proliferative vitreoretinopathy, thus representing an indicator of significant risk for evolution of the disease at a very early time. PMID- 9865622 TI - External beam radiation in patients suffering from exudative age-related macular degeneration. A matched-pairs study and 1-year clinical follow-up. AB - BACKGROUND: The aim of this prospective study was to ascertain whether external beam irradiation is effective in patients with subretinal neovascularization (SRN) due to age-related macular degeneration (AMD). METHODS: All patients had subfoveal SRN due to AMD as verified by fluoresceinangiography. Two hundred and eighty-seven patient-eyes were treated by external beam irradiation (total dose of 16 Gy in 2-Gy fractions, 5 times a week) from January 1996. The analysis was restricted to those 73 patients with a minimum follow-up of 11 months. Eighteen patients with subfoveal SRN who refused treatment served as control group (CG). 18 patients of the treatment group (TG) were matched for visual acuity, refraction and extent of SRN. The statistical analysis was performed with the unpaired t-test. RESULTS: The mean age of the CG was 73.9 years (range 66.9-81.3 years) and of the TG 75.6 years (range: 65.7-80.6 years). The median follow-up was 13.5 months (range 11.9-18.4 months) in the CG and 12.9 months (range 11-13.9 months) in the TG. The initial visual acuity (VA) was 20/80 in both groups. After 7 months the follow-up revealed median VA of 20/400 in the CG and 20/160 in the TG (P=0.0335). The final median VA was 20/400 in both groups, with a range from 20/40 to 20/1000 in the CG and from 20/63 to 20/500 in the TG (P=0.2433). The SRN doubled in size during this time in both groups. CONCLUSION: These results suggest that external beam irradiation applied in 2-Gy fractions 5 times a week slows down the visual loss in exudative AMD for a short time. Nevertheless, the patients' reading vision could not be saved in the long term. PMID- 9865623 TI - Experimental and clinical evaluation of incision size and shape following forceps and injector implantation of a three-piece high-refractive-index silicone intraocular lens. AB - BACKGROUND: Previous studies have demonstrated that incision sizes required for insertion of various foldable intraocular lenses (IOLs) vary according to the IOL/inserter combination. The purpose of this study was to compare incision sizes and wound shape for both forceps and injector implantation of high-refractive index silicone IOLs. METHODS: In fresh human cadaver eyes, limbal corneal tunnel incisions were created, and 12 foldable high-refractive-index silicone (dimethyldiphenylsiloxane) IOLs were inserted in a randomized fashion using either a forceps or an injector. Using incision calipers, internal and external measurements of the tunnel incisions were obtained before and after IOL insertion. Scanning electron microscopy (SEM) was performed on selected corneas following IOL insertion. Additionally, in 12 cataract procedures, the incision sizes following forceps or injector implantation were evaluated intraoperatively. RESULTS: In the experimental setting, the external and internal tunnel widths (in mm) before insertion were 3.05 mm (+/-0.07) and 3.02 mm (+/-0.03), respectively, with the forceps and 3.06 mm (+/-0.04) and 3.01 mm (+/-0.04) with the injector. Following IOL implantation, the external and internal incision sizes were 3.33 mm (+/-0.07) and 3.33 mm (+/-0.04) with the forceps and 3.32 mm (+/-0.08) and 3.33 mm (+/-0.07) with the injector. SEM showed tearing of corneal structures after implantation through the smallest possible incisions with both devices. In the clinical study, the incision sizes before and after implantation were 3.23 mm (+/ 0.10) and 3.36 mm (+/-0.06) with the forceps and 3.11 mm (+/-0.08) and 3.21 mm (+/-0.10) with the injector. CONCLUSIONS: This study confirms that high refractive-index silicone IOLs provide one of the smallest currently achievable incisions before and after IOL implantation. In clinical use, the new injector reduces the mean incision size required by approximately 0.1 mm. PMID- 9865624 TI - Immunohistochemical localization of Na+/Ca2+ exchanger in human retina and retinal pigment epithelium. AB - BACKGROUND: Impaired calcium (Ca2+) metabolism has been implicated in the pathogenesis of various ocular diseases, suggesting that regulation of Ca2+ homeostasis in the retina is of major significance for normal function. There are known families of transport proteins that can catalyze net Ca2+ efflux across the plasma membrane, one of which is the Na+/Ca2+ exchanger. Using immunohistochemistry, we have investigated the human retina and retinal pigment epithelium (RPE) for the presence and distribution of the so-called cardiac-type Na+/Ca2+ exchanger. METHODS: Paraffin sections of ten eyes enucleated for various disease processes were incubated according to the ABC method with a polyclonal antibody (pi) produced against the canine cardiac sarcolemmal Na+/Ca2+ exchanger. The reaction was visualized with aminoethylcarbazole. RESULTS: There was a positive reaction with anti-Na+/Ca2+ exchanger in the retina and RPE in all eyes, but the labeling varied among the different specimens. In neural retina, staining was most intense in Mueller cells, in cells of the inner nuclear layer, and in cone inner segments. Immunoreactivity was less pronounced in ganglion cells, nerve fibers, the outer nuclear layer and in rod inner segments. Outer segments appeared mostly negative. In the RPE, positive staining was present but the intensity of staining varied both within and among the specimens. Reactive RPE cells revealed the most intense labeling. CONCLUSION: Na+/Ca2+ exchanger of the cardiac type is present in human retina and RPE. The variation in immunoreactivity among the different specimens may reflect the different diseases of these eyes and their different metabolic states. A specific relation between certain diseases and malfunction of the Na+/Ca2+ exchanger could have a major impact on therapeutic regimens. PMID- 9865625 TI - Rapid increase in cytosolic calcium ion concentration mediated by acetylcholine receptors in cultured retinal neurons and Muller cells. AB - PURPOSE: This study was conducted to detect the presence of muscarinic or nicotinic receptors in cultured retinal neurons and Muller cells. METHODS: Pure Muller cell cultures and cocultures of retinal neurons and Muller cells were used; the former, obtained from adult rabbit retinas, and the latter, retinal neurons from neonatal rats, were cocultured with Muller cells. Intracellular calcium ion concentration ([Ca2+]i) following the administration of acetylcholine, a cholinesterase inhibitor (trichlorfon), nicotine or muscarinic agonist with or without a receptor antagonist was monitored using the calcium ion indicator, fura-2. RESULTS: Acetylcholine and trichlorfon induced rapid increase in [Ca2+]i in half of either cell type. Trichlorfon induced positive response in coculture but not in the pure Muller cell cultures. This positive response was blocked only partially in the presence of atropine. Approximately 30-40% of neurons responded to nicotine at 5 microM, which was significantly blocked by alpha-bungarotoxin at 50 nM. No response to nicotine could be detected in Muller cells. Approximately 50% of neurons responded to muscarine at 50 microM, but 500 microM was required for the formation of calcium transients in 50% of Muller cells. The muscarine inducement of rapid increase in [Ca2+]i was blocked by atropine. The agonist of M1 (a muscarinic receptor subtype), McN-A-343, at 0.5 microM induced the most significant and rapid increase in [Ca2+]i both in neurons and Muller cells. McN-A-343 administration at 0.05 microM induced positive response in half the neurons, but only in approximately 10% of Muller cells. Such positive response was not observed following preincubation with the M1 antagonist, pirenzepine, at 50 microM. CONCLUSIONS: Cocultured retinal neurons enhance the release of acetylcholine following anticholinesterase administration, and approximately half the neurons were found to possess muscarinic and nicotinic receptors. However, Muller cells appeared to possess only the less sensitive muscarinic receptor. Muscarinic receptor subtypes on either type of cell contained at least M1. PMID- 9865626 TI - Unilateral pigmentary degeneration of the retina associated with heterochromia iridis. AB - BACKGROUND: For the past 5 years, a 56-year-old patient has been displaying monocular progressive pigmentary changes in the left eye. Heterochromy of the left eye has been known since childhood. The other eye is clinically and functionally normal. The patient was adopted and he has no children. Therefore, we have no family history. METHODS: The patient was examined clinically and by means of electroretinography, electrooculography, perimetry, computer tomography, pulsatile ocular blood flow (POBF) measurement, serology and Doppler sonography. RESULTS: Electrophysiology displayed a considerable reduction of scotopic and photopic ERGs, a reduced dark-through, and a reduced light-rise in the left eye, whereas the fellow eye was normal. The visual field was limited to 5 deg around the fixation point, and a peripheral crescent-shaped arch encircled the temporal inferior quadrant concomitant to the pigmentary changes. By computer tomography and Doppler sonography a vascular affection was excluded. The left eye displayed lower POBF values. All serological tests were found negative. CONCLUSION: The clinical picture and negative exclusion criteria indicate a unilateral retinitis pigmentosa. However, with regard to the literature an unequivocal diagnosis can only be made upon hereditary evidence. PMID- 9865627 TI - Human cytomegalovirus infection in a retinoblastoma cell line in vitro. AB - BACKGROUND: The focus of these studies was to determine whether the Y79 human retinoblastoma cell line could function as a good in vitro model system for studying human cytomegalovirus (HCMV) infection. METHODS: Y79 cells were exposed to an HCMV mutant carrying a LacZ gene, and the resulting beta-galactosidase expression in infected cells was assessed by flow cytometry. The extent to which the three classes of viral gene products immediate early, early, and late proteins - were expressed was analyzed by immunohistochemical staining and Western blotting. Infected Y79 cells were also co-cultivated on human foreskin fibroblast (SF cell) cultures to recover virus. RESULTS: Infection of Y79 cells with the virus resulted in beta-galactosidase expression as detected by flow cytometric analysis. Immunohistochemical staining revealed that a portion of Y79 cells expressed antigens reactive to monoclonal antibodies against immediate early, early, and late HCMV proteins. The 43-kDa early gene product was also detected by Western blotting. Infected Y79 cells co-cultivated on SF cell cultures yielded infectious foci, which turned blue following X-gal staining, demonstrating productive HCMV infection in the Y79 cells. CONCLUSION: These results demonstrate that while HCMV can productively infect Y79 cultures, it does so in a highly inefficient manner, leading these authors to conclude that this cell line does not provide a particularly good model system to study HCMV infection. PMID- 9865628 TI - Flash pump dye laser treatment of laryngeal papillomas. AB - To evaluate the feasibility and safety of the flash pump dye (FPD) laser for the treatment of laryngeal papillomas, we performed a prospective nonrandomized trial comparing FPD and carbon dioxide (CO2) laser treatment of laryngeal papillomas in a tertiary care children's hospital. Nine patients from 2 to 20 years of age with severe recurrent laryngeal papillomas were enrolled in the study. All patients underwent CO2 laser debulking of papillomas on the left hemilarynx, with 8 W continuous or pulsed energy. The right hemilarynx was treated with the FPD laser with 12 to 77 pulses at 8 to 12 J, through a custom-designed 90 degrees firing fiber. An end point of blanching and purpura of the papillomas was used. The main outcome measures were the safety and ease of operation with the FPD laser, and decreased papilloma size based on visual inspection at 2 weeks postoperative and at the next laser procedure. Seven patients were irradiated I time with an FPD laser, and 2 patients 2 times. No intraoperative complications were noted. One patient developed early postoperative stridor. No patients described more discomfort, and 5 patients described their voice as being the same as or better than it was after prior CO2 laser procedures. Five patients had a 90% or more decrease in size of papillomas on the FPD-irradiated side 2 weeks postoperatively. Three patients had approximately a 50% reduction. A treatment effect was noted in all patients, and was similar to the results noted on the CO2 laser-treated side. Early results with FPD laser treatment of laryngeal papillomas suggest the protocol is relatively safe and feasible. Long-term results are pending. Because the FPD laser coagulates rather than vaporizing tissue, potential advantages may include decreased scarring relative to CO2 laser treatment, and improved patient and operator safety. PMID- 9865629 TI - Use of Montgomery T-tube in laryngotracheal reconstruction in children: is it safe? AB - Montgomery T-tubes are often used in adult patients; however, they are less commonly used in the pediatric age group. The purpose of this report is to describe our experience with this stent in pediatric laryngotracheal reconstruction. A retrospective chart review was performed to identify early and late complications. Nutritional assessments were made on the basis of weight percentile comparisons at the time of surgery and at the end of the stenting period. From 1990 to 1997, the Montgomery T-tube was utilized in 26 children undergoing 36 airway reconstruction procedures (21 laryngotracheoplasties and 15 cricotracheal resections). The upper limb of the tube extended above the level of the glottis in all patients. The patients ranged in age from 2.4 to 17.9 years. The duration of stenting ranged from 2 weeks to 23 months. Three patients (11.5%) had significant aspiration and did not improve following diet modifications and swallowing therapy, requiring tube feedings. One patient had postoperative subcutaneous emphysema that resolved spontaneously. Three patients required early removal of the stent due to an inability to tolerate plugging. Granulation tissue above the upper limb of the stent during the stenting period was noted after 6 laryngotracheal reconstruction procedures (16%). Only 1 patient fell off his growth curves during the period of stenting. There were no deaths in this series, and no emergent procedures were required. Postoperative and home care and management of complications are discussed. Our experience indicates that Montgomery T-tubes can be utilized relatively safely in children, providing that postoperative and home care are meticulous. PMID- 9865630 TI - Evaluation of gastroesophageal reflux in patients with laryngotracheal stenosis. AB - To report the incidence of pharyngeal acid reflux events in patients with laryngotracheal stenosis (LTS), we studied 12 patients with LTS and 34 healthy volunteers. All patients and controls underwent ambulatory 24-hour 3-site pH monitoring. In ambulatory pH monitoring, pH was recorded at manometrically determined sites of the pharynx, proximal esophagus, and distal esophagus. For all 3 sites, a pH value below 4 that was not related to the time of oral intake or belching was considered an acid reflux event. Eight of the 12 LTS patients exhibited pharyngeal acid reflux events. In the control group, pharyngeal acid reflux events were documented in 7 subjects. In between-group comparison, the number of reflux episodes and the percent acid exposure time in the pharynx were greater in LTS patients than in controls. Reflux parameters of the proximal and distal esophagus in LTS patients were similar to those of controls. The incidence of pharyngeal acid reflux events in LTS patients was higher than that in controls. It is suggested that identification and treatment of gastroesophageal reflux in patients will significantly simplify and improve the results of treatment for LTS. PMID- 9865631 TI - Thirty-eight-year evaluation of a surgical technique to protect the external branch of the superior laryngeal nerve during thyroidectomy. AB - The purpose of this study is to evaluate a surgical technique for the preservation of the function of the external branch of the superior laryngeal nerve (EBSLN) during total thyroid lobectomy and total thyroidectomy. Permanent injury to this nerve can be a disaster, especially in singers and professional speakers who depend on control of pitch, and a clear and forceful voice. Voice changes may be either obvious or subtle. For better voice analysis, a detailed questionnaire is necessary. Thus, the evaluation in this study is based on laryngoscopy of 934 nerves in 675 patients and detailed subjective voice evaluation of 66 patients during the last 2 years of the study. There are 2 surgical principles. First, the EBSLN is not routinely exposed; the distal 1.5 to 2.0 cm (critical area) of the superior thyroid vessels are carefully dissected, exposed, and ligated, preferably independently. Careful observation ensures that the EBSLN is not included in the ligature. Second, the ultimate evaluation of the surgical technique is the effect of voice changes on the patient's lifestyle. Laryngoscopy of 934 nerves found bowing, temporary in 4 patients and permanent in 1 patient (limited follow-up). Of the 66 patients with voice evaluations, 14 had changes: 9 had temporary changes, while 5 had permanent changes. In these 14 patients, voice changes had no effect on lifestyle in 13, and the effects were indeterminate in 1 (unavailable for follow-up). The estimated deleterious effect of voice changes on lifestyle is no greater than 1.5% of the 66 patient responders. PMID- 9865632 TI - Reticular fibers in the vocal fold mucosa. AB - Light and electron microscopic investigations of the reticular fibers (RFs) in the vocal fold mucosa were carried out on excised human adult larynges. The results are summarized as follows. Reticular fibers were found in the superficial and intermediate layers of the lamina propria of the vocal fold mucosa. They were most abundantly discovered around the vocal fold edge, and they decreased toward the superior and inferior portions of the vocal folds. The RFs were composed of slender fibrils, about 40 nm in diameter, and having cross-bands with a periodicity of about 67 nm. They were found in close association with the basal lamina of the epithelium and blood vessels. The slender fibrils of the RFs did not form any bundles, but branched and anastomosed. The RFs formed delicate 3 dimensional networks, and the spaces among the fibers were relatively large. Glycoprotein and glycosaminoglycan (proteoglycan) were situated around the RFs and in the spaces among the fibers. Elastic fibers were located in the spaces among the RFs. The 3-dimensional structure of the RFs in the vocal fold mucosa, first demonstrated in this study, appears to be one of the key components of the structural maintenance and viscoelasticity of the vibrating vocal fold tissue. PMID- 9865633 TI - Calcitonin gene-related peptide-like immunoreactive motoneurons innervating the canine intrinsic laryngeal muscles. AB - The percentage of calcitonin gene-related peptide (CGRP)-immunoreactive motoneurons in the nucleus ambiguus innervating the intrinsic laryngeal muscles of colchicine-treated dogs was examined by using cholera toxin B subunit-gold (CTBG) as a retrograde tracer and by immunohistochemistry. Neurons that were labeled with CTBG from the cricothyroid muscle were located in the ventromedial division of the rostral part of the nucleus ambiguus, and the ratio of CGRP positive neurons was 93.0%. Neurons labeled with CTBG from the thyroarytenoid muscle were located in the dorsal division of the medial part of the nucleus ambiguus, and the percentage of neurons with CGRP was 71.4%. Neurons labeled with CTBG from the posterior cricoarytenoid muscle were located in the ventral division of the medial part of the nucleus ambiguus, and the percentage of CGRP positive neurons was 85.5%. These findings suggest that the innervation and/or the neurotrophic mechanism involving CGRP for each intrinsic laryngeal muscle is different. PMID- 9865634 TI - Endoscopic transseptal sphenopalatine artery ligation for intractable posterior epistaxis. AB - The rigid nasal endoscope was used through a transseptal approach to reach the sphenopalatine foramen and to ligate its artery in 9 patients with intractable posterior epistaxis. Immediate and complete cessation of the bleeding uniformly occurred, except in 1 case, in which there was persistent bleeding on endoscopic examination of the nasal cavity at the end of the procedure. The ligature was checked and the artery was reclipped. Thereafter, the patient's recovery was uncomplicated and free of further epistaxis. Endoscopic transseptal sphenopalatine artery ligation offers a reliable option in the treatment of intractable posterior epistaxis. The submucoperiosteal dissection reduces bleeding, shortens operation time, and allows relatively easy identification of the sphenopalatine foramen. The procedure allows direct positive control of the major vessel supplying the posterior nasal cavity. It avoids the complications associated with transantral and pterygopalatine fossa surgery. PMID- 9865635 TI - TNM glottic: role of the vocal muscle, arytenoid cartilage, and inferior paraglottic space in impaired vocal cord mobility (T2). An embryological and clinical study. AB - The TNM classification is unsatisfactory regarding the impaired mobility of the vocal cord and/or the arytenoid. The authors have studied the intrinsic laryngeal musculature, with particular attention to the medial thyroarytenoid muscle, the lateral thyroarytenoid muscle, and the inferior paraglottic space. The study was conducted on serial sections of 4 embryonal and 9 fetal larynges at different stages of development. The embryological observations showed that the lateral and medial thyroarytenoid muscles have different maturation times and probably different functions: phonatory and sphincteric. The authors have found that the inferior paraglottic space is situated between these 2 muscles. It would represent a path of diffusion for glottic cancer and play a central role in impaired vocal cord and/or arytenoid mobility, according to the degree of diffusion. Embryological observations have suggested the existence of laryngeal morphofunctional units belonging to the subsites of the glottic region and related to tumoral spreading. PMID- 9865636 TI - Treatment of benign positional vertigo using heels-over-head rotation. AB - Particle repositioning, using either bedside techniques or whole-body manipulation devices, has been used effectively to treat benign positional vertigo (BPV). We assessed the efficacy of particle repositioning using a device designed and built specifically to treat BPV that rotated patients 360 degrees heels-over-head in their sagittal body plane while their heads were turned to align the posterior semicircular canal with the plane of rotation. Eye movements were monitored during the maneuver by an infrared video recording system that allowed subsequent review of the induced nystagmus. Our results indicate that 1) heels-over-head rotation is an extremely efficacious procedure for treating patients with BPV and 2) the pattern of nystagmus during repositioning is consistent with the theory that free-floating debris is highly likely to account for BPV. PMID- 9865637 TI - Neurotological auditory brain stem response findings in human immunodeficiency virus-positive patients without neurologic manifestations. AB - Neurologic manifestations of human immunodeficiency virus (HIV) infection, rather than being a late complication of the disease, are principally correlated with the early central nervous system (CNS) localization of HIV. The CNS may be infected in the early stages of acquired immunodeficiency syndrome (AIDS) without evidence of neurologic disorders. Evoked potentials (visual, auditory, and somatosensory) and electronystagmographic test batteries have proven to be very sensitive in showing subclinical CNS disorders due to HIV. In this study, auditory brain stem response (ABR) and electronystagmographic test battery findings (smooth pursuit, saccades, caloric test) were performed in 29 neurologically asymptomatic, HIV-positive subjects at different stages of the disease. Compared to results of a control group, the ABR latencies of waves V, I, and III and interpeaks I-V and III-V were significantly increased in HIV patients. The same parameters did not differ significantly among the stages of the disease. In HIV-positive subjects, the accuracy of saccades was significantly reduced, while latency was normal. The velocity and the gain of pursuit were significantly reduced in HIV-positive patients, and 15 of 29 patients showed corrective saccades. Caloric tests revealed qualitative nystagmus abnormalities in 82% of HIV patients, while quantitative parameters were normal. The present results confirm that CNS involvement by HIV occurs early in the course of the disease. In particular, HIV does not seem to affect the labyrinth or the eighth cranial nerve, as demonstrated by the normality of the I-III value of the ABR and of the quantitative parameters of the caloric responses, but it does appear to involve the brain stem acoustic pathways, pontocerebellar pathways, and supratentorial areas. PMID- 9865638 TI - Speech and voice characteristics after near-total laryngectomy. A preliminary prospective study. AB - Selected characteristics were prospectively compared in the speech and voice of 10 patients managed with near-total laryngectomy. Tape-recorded speech samples were measured for durational features with a stopwatch. Acoustic features were analyzed with the Computerized Speech Lab and the Multidimensional Voice Program from Kay Elemetrics. Speech and voice parameters recorded preoperatively were compared with the parameters recorded postoperatively. Our data corroborate the prior reported good quality of the voice and speech after near-total laryngectomy. PMID- 9865639 TI - Kimura's disease: two case reports and a literature review. AB - Kimura's disease (KD) is an allergic, inflammatory disorder of unknown cause. The typical presentation is in a young Oriental male with nontender subcutaneous swellings in the head and neck region, lymphadenopathy, peripheral eosinophilia, and elevated serum IgE. Many patients with KD also develop renal involvement. Treatment options range from conservative observation for asymptomatic patients to surgical excision, steroid therapy, and radiotherapy for symptomatic patients. Two cases of KD are presented. These emphasize not only the variability of disease presentation, but also the pitfalls of therapy leading to frequent recurrences. The literature is reviewed, summarizing patient presentation, differential diagnosis, current theories on causation, and therapy. A treatment protocol is also proposed. PMID- 9865640 TI - Retropharyngeal abscess and Epstein-Barr virus infection in children. AB - A 6-year retrospective chart review was conducted of all pediatric patients with the diagnosis of retropharyngeal abscess (RPA) at the Columbia-Presbyterian Medical Center. The charts were examined for signs, symptoms, and serologic findings consistent with recent acute Epstein-Barr virus (EBV) infection. Of the 7 patients we studied, 4 had elevated immunoglobulin G antibodies that were consistent with recent acute EBV infection, and 3 patients were not tested. The 4 patients with positive titers also presented with signs typical for acute EBV infection, including fever (3/4), lymphadenopathy (4/4), and pharyngitis (4/4). Of note, the 2 patients who were older than expected for RPA (ages 18 and 11) had the most severe infections. We submit that there may be a role for EBV infection in the pathogenesis of RPA formation in children. Increasing age may correlate with a more severe infection in this setting. PMID- 9865641 TI - Cochlear otosclerosis: the current understanding. PMID- 9865642 TI - Synovial sarcoma of the larynx and hypopharynx. AB - Synovial sarcoma represents a mesenchymal malignancy of unknown histogenesis that most often occurs in the lower limbs of young adults. The head and neck region is a relatively rare location, in which the hypopharynx and larynx are, respectively, the most and least often affected anatomic sites. Histologically, synovial sarcomas are classified into monophasic and biphasic variants. Immunohistochemistry plays a major part in the differential diagnosis, enabling the demonstration of epithelial differentiation. Both monophasic and biphasic synovial sarcomas are characterized cytogenetically by the reciprocal translocation t(X;18)(p11.2;q11.2) between chromosomes X and 18. Two cases of synovial sarcoma arising in the larynx and in the hypopharynx and in which cytogenetic analysis detected a diagnostic t(X;18) chromosome aberration are reported here. PMID- 9865643 TI - Comparison of wound culture and bronchial lavage in the severely burned child: implications for antimicrobial therapy. AB - BACKGROUND: The relationship of the burn wound flora to microbial pathogens in the tracheobronchial tree has important implications for antimicrobial therapy in the severely burned patient. Management of septic complications is bolstered by surveillance quantitative wound cultures (QWC) and bronchial lavage fluid (BLF) cultures. OBJECTIVES: To compare the organisms present in BLF with those found in QWC and to determine if QWC can predict BLF results. DESIGN: Results of BLF cultures from all patients who underwent bronchial lavage from January 1, 1996, to December 31, 1996, at our institution were compared with QWC data from the same date. Criteria for a positive match included an identical antibiotic susceptibility pattern and biotype. Match rates were calculated qualitatively and quantitatively. RESULTS: In 30 (48%) of the 62 BLF cultures, there was a match between the organism identified in the BLF and the QWC. When strict quantitative criteria were applied, the match rate was only 9 (14%) of 62. Burn size and inhalation injury had no significant effect on match rate. CONCLUSIONS: Whereas the microbial pathogens were similar in the QWC and BLF, linear regression showed no value of QWC in predicting BLF culture results. The difference between qualitative and quantitative match rates suggests cross-colonization between the burn wound and tracheobronchial tree, but little to no cross-infection. The QWC and BLF cultures must be performed independently in determining antimicrobial specificity in the burned patient. PMID- 9865644 TI - Dehydroepiandrosterone: an inexpensive steroid hormone that decreases the mortality due to sepsis following trauma-induced hemorrhage. AB - BACKGROUND: Recent studies suggest that male sex steroids play a role in producing immunodepression following trauma-hemorrhage. This notion is supported by studies showing that castration of male mice before trauma-hemorrhage or the administration of the androgen receptor blocker flutamide following trauma hemorrhage in noncastrated animals prevents immunodepression and improves the survival rate of animals subjected to subsequent sepsis. However, it remains unknown whether the most abundant steroid hormone, dehydroepiandrosterone (DHEA), protects or depresses immune functions following trauma-hemorrhage. In this regard, DHEA has been reported to have estrogenic and androgenic properties, depending on the hormonal milieu. OBJECTIVE: To determine whether administration of DHEA after trauma-hemorrhage has any salutary or deleterious effects on immune responses, and whether it improves the survival of animals subjected to subsequent sepsis. DESIGN: Male C3H/HeN mice underwent laparotomy (ie, trauma induced) and hemorrhagic shock (blood pressure, 35+/-5 mm Hg for 90 minutes) followed by fluid resuscitation, or sham operation. The animals then received 100 mg of DHEA (4 mg/kg) or propylene glycol (hereafter referred to as vehicle). At 24 hours after trauma-hemorrhage and resuscitation, the animals were killed and blood, spleens, and peritoneal macrophages were harvested. Splenocyte proliferation and interleukin (IL) 2 release and splenic and peritoneal macrophage IL-1 and IL-6 release were determined. In a separate set of experiments, sepsis was induced by cecal ligation and puncture at 48 hours after trauma-hemorrhage and resuscitation. For those studies, the animals received vehicle, a single 100-microg dose of DHEA, or 100 microg/d DHEA for 3 days following hemorrhage and resuscitation. Survival was monitored for 10 days after the induction of sepsis. RESULTS: Administration of DHEA restored the depressed splenocyte and macrophage functions at 24 hours after trauma-hemorrhage. Moreover, daily administration of DHEA for 3 days significantly increased the survival of animals subjected to subsequent sepsis (P=.01). CONCLUSION: The finding that DHEA markedly improves the depressed immune functions and survival of animals subjected to subsequent sepsis suggests that short-term treatment with DHEA after trauma-hemorrhage is a safe and novel approach for preventing immunodepression and for decreasing the mortality rate due to subsequent sepsis. PMID- 9865646 TI - The pivotal role of adrenomedullin in producing hyperdynamic circulation during the early stage of sepsis. AB - BACKGROUND: Initial cardiovascular responses during sepsis are characterized by hyperdynamic circulation. Although studies have shown that a novel potent vasodilatory peptide, adrenomedullin (ADM), is up-regulated under such conditions, it remains unknown whether ADM is responsible for initiating the hyperdynamic response. OBJECTIVE: To determine whether increased ADM release during early sepsis plays any major role in producing hyperdynamic circulation. DESIGN, INTERVENTION, AND MAIN OUTCOME MEASURE: Synthetic rat ADM (8.5 microg/kg of body weight) was infused intravenously in normal rats for 15 minutes at a constant rate. Cardiac output, stroke volume, and microvascular blood flow in various organs were determined immediately as well as 30 minutes after ADM infusion. At 30 minutes after infusion, plasma ADM level was also measured. In additional groups, rats were subjected to sepsis by cecal ligation and puncture. At 1.5 hours after cecal ligation and puncture, specific anti-rat ADM antibodies were infused, which completely neutralized the circulating ADM. Various hemodynamic variables were measured 5 hours after cecal ligation and puncture (ie, the early stage of sepsis). RESULTS: Cardiac output, stroke volume, and microvascular blood flow in the liver, small intestine, kidney, and spleen increased, and total peripheral resistance decreased 0 and 30 minutes after ADM infusion. In addition, plasma levels of ADM increased from the preinfusion level of 92.7+/-5.3 to 691.1+/-28.2 pg/mL 30 minutes after ADM infusion, which was similar to ADM levels observed during early sepsis. Moreover, 5 hours after the onset of sepsis, cardiac output, stroke volume, and microvascular blood flow in various organs increased and total peripheral resistance decreased. Administration of anti-ADM antibodies, however, prevented the occurrence of the hyperdynamic response. CONCLUSIONS: The results suggest that increased ADM production and/or release plays a major role in producing hyperdynamic responses during early sepsis. Since our previous studies have shown that vascular responsiveness to ADM decreases in late sepsis, maintenance of ADM vascular responsiveness by pharmacological agents during the course of sepsis may prevent transition from the hyperdynamic to the hypodynamic state. PMID- 9865645 TI - A trial of ciprofloxacin and metronidazole vs gentamicin and metronidazole for penetrating abdominal trauma. AB - OBJECTIVES: To determine whether a combination of ciprofloxacin hydrochloride and metronidazole hydrochloride would be as effective or more effective than a combination of gentamicin sulfate and metronidazole hydrochloride for preventing infection in patients with penetrating abdominal trauma, to evaluate the factors associated with increased risk of infection, and to determine the serum peak and trough levels of gentamicin with the dosage regimen of 2.5 mg/kg every 12 hours. DESIGN: Randomized double-blind study. SETTING: Level I trauma center. PATIENTS: Eighty-four patients with penetrating intra-abdominal injuries (gunshot wound, 69; stab wound, 15) thought to require laparotomy. INTERVENTIONS: The patients were randomized during treatment in the emergency department to be given a combination of ciprofloxacin hydrochloride, 400 mg every 12 hours, and metronidazole hydrochloride, 500 mg every 6 hours, or a combination of gentamicin sulfate, 2.5 mg/kg every 12 hours, and metronidazole hydrochloride, 500 mg every 6 hours. RESULTS: Of 68 patients with intra-abdominal injuries who could be observed for at least 48 hours after laparotomy, posttraumatic infections developed in 12 (18%), and nosocomial infections developed in 6 (9%). The incidence of posttraumatic infections in patients who were given gentamicin and metronidazole (5/33 [15%]) was not significantly lower than the incidence in patients who were given ciprofloxacin and metronidazole (7 of 35 [20%]; P=.75). The presence of any infection increased the mean+/-SD length of hospital stay from 8.7+/-3.5 days to 23.3+/-10.9 days and increased the mean+/-SD hospital charges from $24 507+/-$9860 to $104920+/-$49083 (P<.001). Univariate analysis showed the factors most significantly associated with infection were as follows: (1) the use of blood transfusions (P<.001), (2) the penetrating abdominal trauma index of 35 or more (P<.002), (3) injury to the colon requiring a colostomy (P=.004), and (4) a trauma score of less than 12 (P<.02). Multivariate analysis showed the only significant factor was the receipt of blood transfusions (F=10.165; P<.005). CONCLUSIONS: Ciprofloxacin and gentamicin, each in combination with metronidazole, were equivalent in their ability to prevent infections after penetrating abdominal trauma; other factors, especially the receipt of blood transfusions, had much more effect on the incidence of infection. Infection greatly increases the length of hospital stay and hospital charges. The use of an increased dosing regimen of 2.5 mg/kg every 12 hours of gentamicin sulfate was effective at obtaining a therapeutic peak serum concentration. PMID- 9865647 TI - Reduction in neutrophil cell surface expression of tumor necrosis factor receptors but not Fas after transmigration: implications for the regulation of neutrophil apoptosis. AB - OBJECTIVES: To test the hypothesis that loss of polymorphonuclear neutrophil tumor necrosis factor alpha (TNF-alpha) receptors during transmigration renders the exudate neutrophil refractory to TNF-alpha-mediated stimulation of apoptosis; and to investigate the surface expression of Fas on both circulating and exudate neutrophils. DESIGN: A prospective cohort study. SETTING: Surgical laboratory of a tertiary care hospital. PARTICIPANTS: Twenty-one healthy human volunteers. INTERVENTIONS: All subjects had circulating neutrophils and exudate neutrophils collected by venipuncture and skin window methods, respectively. MAIN OUTCOME MEASURES: Circulating and exudate neutrophils were incubated in culture medium (1.0x10(6) neutrophils per milliliter) alone or with TNF-alpha (100 ng/mL). Apoptosis was evaluated by flow cytometry (annexin V-fluorescein isothiocyanate and propidium iodide). Tumor necrosis factor alpha-phycoerythrin and anti-human Fas-fluorescein isothiocyanate were used to evaluate neutrophil TNF-alpha receptors and surface expression of Fas. RESULTS: Exudate neutrophils had a significant delay in apoptosis rates when compared with circulating neutrophils. The percentage of neutrophils expressing TNF-alpha receptors was significantly diminished after exudation (80%+/-15% vs 33%+/-9%; P<.001), as was the median channel number of TNF-alpha phycoerythrin fluorescence (8.1+/-1.6 vs 5.2+/-0.5; P=.001). However, the expression of Fas was unchanged after transmigration (percentage positive for Fas: 98.7%+/-0.7% vs 92.8%+/-3.4%, P=.89; Fas antibody fluorescein isothiocyanate median channel fluorescence: 12.2+/-1.1 vs 13.1+/-1.2; P=.80). Exposure of exudate neutrophils to TNF-alpha failed to increase their rate of apoptosis. CONCLUSIONS: Exudate polymorphonuclear neutrophils are confirmed to have delayed apoptosis. Loss of TNF-alpha receptors during transmigration is necessary for neutrophil survival in the extravascular inflammatory milieu. PMID- 9865648 TI - Nuclear factor-kappaB is activated in intestinal mucosa during endotoxemia. AB - BACKGROUND: The transcription factor nuclear factor-kappaB (NF-kappaB) regulates a large number of genes involved in the inflammatory response to critical illness. The intestinal mucosa plays an active role in the inflammatory and metabolic response to sepsis and endotoxemia, but it is not known if NF-kappaB is activated in the mucosa during these conditions. OBJECTIVE: To test the hypothesis that endotoxemia in mice activates NF-kappaB in intestinal mucosa. METHODS: Mice were injected subcutaneously with lipopolysaccharide, 12.5 mg/kg, or a corresponding volume of saline. At various intervals following injection, jejunal mucosa was harvested and nuclear and cytoplasmic fractions were prepared. The nuclear fractions were analyzed by electrophoretic mobility shift assay for NF-kappaB activation and by Western blot analysis for the NF-kappaB subunits p50 and p65. Cytoplasmic fractions were analyzed by Western blotting for the NF kappaB inhibitory proteins IkappaB-alpha and IkappaB-beta. RESULTS: Electrophoretic mobility shift assay showed that NF-kappaB was activated in jejunal mucosa 1 hour after injection of lipopolysaccharide and persisted for at least 4 hours. The NF-kappaB subunits p50 and p65 were present in nuclear fractions of mucosa from endotoxemic mice at the corresponding time points. Cytoplasmic levels of the inhibitory proteins IkappaB-alpha and IkappaB-beta decreased during endotoxemia, and the proteins were nearly absent 60 minutes after injection of lipopolysaccharide. CONCLUSIONS: The results suggest that IkappaB is degraded and NF-kappaB is activated in intestinal mucosa during endotoxemia. The findings support the concept that the intestinal mucosa is an important component of the inflammatory response to sepsis and endotoxemia. PMID- 9865649 TI - Skeletal muscle phosphocreatine depletion depresses myocellular energy status during sepsis. AB - OBJECTIVE: To determine the effects of phosphocreatine (PCr) depletion on myocellular energetics. DESIGN: Randomized controlled study. SETTING: University laboratory. MATERIALS: Thirty-eight adult male Wistar rats (110-121 g). METHODS: The poorly metabolized creatine analogue beta-guanidinopropionic acid, (beta-GPA, 2% of a gel diet) was fed to the rats for 14 days to replace (75%) endogenous PCr stores before cecal ligation and puncture (CLP). Rats were randomized to receive sham operation and gel diet (sham-gel group [n=10]), sham operation and beta-GPA diet (sham-beta-GPA group [n=9]), CLP and gel diet (CLP-gel group [n=10]), and CLP and beta-GPA diet (CLP-beta-GPA group [n=9]). On day 14, all animals underwent operation. Twenty-four hours later, in vivo phosphorus 31-labeled magnetic resonance spectroscopy (31P-MRS) of the gastrocnemius muscle was performed. Muscle samples were collected to determine enzyme activities of beta hydroxyacyl-CoA dehydrogenase, phosphofructokinase, citrate synthase, and the metabolites adenosine triphosphate (ATP), PCr, inorganic phosphate, and creatine. Free adenosine diphosphate levels, the phosphorylation potential, and free energy change of ATP hydrolysis were then calculated. RESULTS: All animals undergoing CLP but no controls had positive results of blood cultures. Although sham-beta GPA animals had altered bioenergetics, CLP-beta-GPA rats experienced a greater deterioration of energy state compared with CLP-gel controls. Glycolytic and oxidative enzyme activities were not significantly different between groups and therefore could not explain the observed differences. CONCLUSIONS: There is an overall decrease in energy availability during sepsis, which is worsened by PCr depletion. These changes support the contention that PCr plays an important role as an ATP buffer during systemic infection. PMID- 9865650 TI - The influence of human endotoxemia on CD95-induced apoptosis. AB - BACKGROUND: The responses of monocyte and neutrophil tumor necrosis factor receptor type 1 (TNFR-1) and TNFR-2 during systemic inflammation have been described previously. Several other members of the TNFR superfamily also appear to have regulatory roles in immunocyte function, including apoptosis. However, the response of these other receptor members, such as CD95, to systemic inflammation is unclear. OBJECTIVES: To compare the response of CD95 with that of TNFR during systemic inflammation and to assess the influence of the inflammatory milieu on CD95 function. SETTING: Adult clinical research center of a university hospital. SUBJECTS AND METHODS: Five healthy male subjects were administered intravenous endotoxin (2 ng/kg), and systemic response was measured by cytokine analysis and receptor expression assays during a 48-hour period. CD95 function during systemic inflammation was assessed using a Jurkat cell bioassay for degree of apoptosis. RESULTS: Monocyte and neutrophil CD95 expression exhibited changes parallel to that of TNFR following endotoxin injection. In contrast to soluble TNFR, which was transiently elevated during endotoxemia, soluble CD95 levels remained unchanged from baseline. Jurkat cells incubated in normal and post endotoxin serum samples equally exhibited less than 10% spontaneous apoptosis. No soluble CD95 ligand was detectable in experimental human endotoxemia. CONCLUSIONS: Cell-associated CD95 exhibited changes parallel to its receptor family member TNFR following endotoxin administration. Soluble CD95 is present in human serum samples, but the levels remained unchanged following endotoxin administration. No soluble CD95 ligand activity was detectable by enzyme-linked immunosorbent assay or by functional assay. The potential protective role of soluble CD95 in human serum samples against CD95 ligand-induced apoptosis remains to be defined. PMID- 9865651 TI - Effect of hypoxia on the hematopoietic and immune modulator preprotachykinin-I. AB - OBJECTIVES: To determine the effect of hypoxia on bone marrow mononuclear cells (BMMCs) and their ability to proliferate into granulocyte-macrophage colony forming units (CFU-GMs) and erythroid burst-forming units (BFU-Es) and to determine the role of the neuroimmune and hematopoietic mediator, substance P. DESIGN: Controlled in vitro study. SETTING: University research laboratory. MATERIALS: Bone marrow aspirates were obtained from the posterior iliac crests of healthy volunteers after obtaining informed consent. INTERVENTIONS: The BMMCs were divided into the following groups: (1) normoxia, (2) two hours of hypoxia, and (3) six hours of hypoxia. Additional BMMCs were purified before the period of hypoxia, while others were incubated with neurokinin (NK) receptor antagonists. In other experiments, bone marrow stroma was grown to confluence and randomized to the following groups: (1) normoxia, (2) hypoxia, (3) normoxia and interleukin (IL) 1, and (4) hypoxia and IL-1. All groups were cultured for 2, 6, 12, or 24 hours. MAIN OUTCOME MEASURES: The formation of CFU-GMs and BFU-Es was measured after 10 to 14 days of incubation of the BMMCs. The messenger RNA of the preprotachykinin-I (PPT-I) gene and the NK-1 and NK-2 receptors was detected by using semiquantitative reverse transcriptase-polymerase chain reaction or Northern blot analysis on bone marrow stroma. The immunoreactivity of substance P in bone marrow stroma was measured by competitive enzyme-linked immunosorbent assay. RESULTS: Hypoxia resulted in a 110% increase in the number of CFU-GMs and a 78% increase in the number of BFU-E colonies at 6 hours (both P<.05). Elimination of the stromal elements by purification abrogated the increase in colony formation to nonhypoxic levels. Hypoxia induced PPT-I gene expression at 24 hours; however, no PPT-I expression was found in the hypoxic group incubated with IL-1. The receptor, NK-1, was found to be equal in both hypoxic groups; NK-2 was found to have a 4-fold increase in the hypoxia and IL-1 group over the hypoxia alone group and normoxia and IL-1 group. The levels of substance P immunoreactivity were found to be similar in all groups. Incubation of BMMCs with NK receptor antagonists to NK-1 alone or NK-1 and NK-2 decreased the number of CFU-GM and BFU-E colonies similar to the level in controls. CONCLUSIONS: These results indicate that hypoxia has a role in the proliferation and control of CFU GMs and BFU-Es. This control seems to be mediated through the bone marrow stroma and modulated by NK receptors and induction of PPT-I. The neuropeptide, substance P, probably has a role but is clearly not the only mediator involved. PMID- 9865652 TI - Microvascular endothelial cell control of peripheral vascular resistance during sepsis. AB - OBJECTIVE: To determine the endothelial-dependent control of decreased peripheral vascular resistance in skeletal muscle microvessels during evolving sepsis. MATERIALS AND INTERVENTIONS: Acute (4 hours, n=7), established (24 hours, n=7), or chronic (72 hours, n=8) infection was induced in Sprague-Dawley rats (150-175 g) by injecting Escherichia coli and Bacteroides fragilis (1 x 10(9) colony forming units for both) into a subcutaneous sponge. Control animals were injected with an isotonic sodium chloride solution and analyzed at the same time points: (n=6-8 per group). Dilation in response to the topically applied endothelial dependent agonist acetylcholine (ACH) (1 x 10(-9) to 1 x 10(-5) mol/L) was measured in inflow first-order (A1) and precapillary fourth-order (A4) arterioles in cremaster muscle in vivo with videomicroscopy. Acetylcholine dose-response curves were used to determine vascular reactivity by calculating the concentration of ACH necessary to elicit 50% of the maximal dilator response. MAIN OUTCOME MEASURES: In vivo reactivity of striated muscle microvessels to the dilation agonist ACH during acute, established, and chronic infection. RESULTS: A1 vessels were unresponsive to all doses of ACH at all time points. A4 vessels showed an increased dilator response during short-term treatment, which deteriorated over time to depressed dilation during chronic infection. CONCLUSIONS: Precapillary A4 vessels have increased dilator reactivity during early sepsis, which progresses to depressed levels with chronic infection. A1 microvessels remain dilated and are not substantially influenced by endothelial dilator mechanisms initiated by ACH. Maximum dilation of the large A1 vessels appears to contribute to the decrease in peripheral vascular resistance noted during systemic infection. PMID- 9865653 TI - Third-generation cephalosporins and vancomycin as risk factors for postoperative vancomycin-resistant enterococcus infection. AB - OBJECTIVE: To examine use of third-generation cephalosporins (3GCs) alone and in association with vancomycin hydrochloride as a risk factor for vancomycin resistant enterococcus (VRE) infection in surgical patients. DESIGN: Case-control retrospective study analyzing antibiotic use in the 30 days preceding culture of VRE or vancomycin-sensitive enterococcus from an infected site. SETTING: A large tertiary care teaching hospital. PATIENTS: Surgical inpatients with VRE infections between September 3, 1993, and January 29, 1997, were matched with patients with vancomycin-sensitive enterococcus infections. Matches were based on surgical procedure, initial infection site, and immunosuppression. Matches were found for 32 of 50 surgical patients with VRE. Twenty matched pairs of patients were recipients of solid organ transplants. MAIN OUTCOME MEASURES: Multivariate logistic regression analysis was done to examine 3GCs and vancomycin as risk factors for VRE infection. Univariate analysis of use of other antibiotic agents and demographic data was also performed. RESULTS: Multivariate analysis showed significant differences in the use of 3GCs both alone and concurrently with vancomycin. Univariate analysis also showed higher use of metronidazole, concurrent vancomycin and metronidazole, concurrent vancomycin and ceftazidime, and all antibiotics combined in patients with VRE infections. CONCLUSIONS: This matched control study showed that use of 3GCs, alone (P=.05) or concurrently with vancomycin (P=.05), was a risk factor for VRE infection in surgical patients. Judicious administration of third-generation antibiotics is warranted in surgical patients with other risk factors for VRE. PMID- 9865654 TI - Multivariate analysis of 9 disease-associated variables for outcome prediction in patients with sepsis. AB - OBJECTIVE: To assess the ability of 9 clinical or biological variables to predict outcome (survival or nonsurvival) using multiple regression and classification analyses. DESIGN: Prospective, descriptive cohort study with no interventions. SETTING: Surgical intensive care unit of a tertiary care hospital and a medical school research laboratory. PATIENTS: Eighteen patients with a documented source of infection who met currently accepted criteria for sepsis syndrome or septic shock. MAIN OUTCOME MEASURES: Prediction of survival or nonsurvival based on analysis of clinical (Multiple Organ Dysfunction score, Acute Physiology and Chronic Health Evaluation III scores) and biological (plasma levels of cortisol, interleukin 6, interleukin 10, phospholipase A2, soluble tumor necrosis factor receptor p75, and monocyte membrane tumor necrosis factor receptor levels) variables, with comparison of predicted and actual outcomes. RESULTS: Plasma interleukin 6, interleukin 10, and phospholipase A2 concentrations were not significantly (P>.05) different between survivors and nonsurvivors. By standard, forward stepwise, and backward stepwise multiple regression analyses, only monocyte membrane tumor necrosis factor receptor levels measured at the onset of sepsis significantly predicted outcome in all 3 analyses. However, by both standard and backward stepwise analyses, Multiple Organ Dysfunction scores based on evaluation at the onset of sepsis and 24 hours later were also significant predictors of outcome. Classification analysis showed that assignment to outcome group was statistically significant when based on monocyte membrane tumor necrosis factor receptor levels determined at the onset of sepsis or on Multiple Organ Dysfunction scores assessed 24 hours after sepsis was diagnosed. CONCLUSION: Although these findings were based on a relatively small cohort, both multiple regression and classification analyses indicated that only monocyte membrane tumor necrosis factor receptor levels are able to discriminate survivors from nonsurvivors at the onset of sepsis. PMID- 9865655 TI - Hemorrhage exacerbates bacterial translocation at low levels of intra-abdominal pressure. AB - BACKGROUND: It has been shown previously that the adverse cardiopulmonary sequelae of increased intra-abdominal pressure (IAP) are worsened by hemorrhage and resuscitation. Bacterial translocation (BT) to the mesenteric lymph nodes (MLNs), liver, and spleen has also been shown to occur with increased IAP. OBJECTIVE: To investigate the hypothesis that BT associated with elevated IAP would be significantly increased after hemorrhage and resuscitation. MATERIALS AND METHODS: Anesthetized adult male rats had femoral artery and vein catheters placed, and an intra-abdominal catheter placed to measure IAP. Group 1 underwent surgery only and served as controls. Group 2 had IAP raised to 10 mm Hg by infused lactated Ringer's solution for 40 minutes. Group 3 had a 25% hemorrhage, followed by resuscitation by infused lactated Ringer's solution and shed blood. Group 4 first had a 25% hemorrhage, resuscitated using infused lactated Ringer's solution and shed blood, and then had IAP raised to 10 mm Hg by infused lactated Ringer's solution for 40 minutes. All groups were killed after 2 hours, and had MLNs, liver, and spleen harvested for quantitative cultures. RESULTS: Hemorrhage and resuscitation alone did not increase BT to the MLNs, liver, or spleen. An increase in IAP to 10 mm Hg resulted in a significant level of BT to the MLNs and liver on MacConkey II agar (P<.05), and a significant increase in the level of BT only to the liver on trypticase soy agar with 5% sheep's blood (P<.05). Hemorrhage and resuscitation did increase the level of BT to the liver and spleen when IAP was increased to 10 mm Hg (P<.05). CONCLUSIONS: In this model, hemorrhage and resuscitation alone did not increase BT to the MLNs, liver, or spleen. However, hemorrhage and resuscitation increased BT to the liver and spleen when IAP was increased to 10 mm Hg. This supports the concept that prior hemorrhage and resuscitation exacerbates the effects of increased IAP. PMID- 9865656 TI - Fasciotomy, chronic venous insufficiency, and the calf muscle pump. AB - OBJECTIVE: To test the hypothesis that fasciotomy may impair the function of the calf muscle pump, which in turn could result in the development of chronic venous insufficiency. DESIGN: A cohort study of patients with a history of lower extremity fasciotomy. SETTING: An urban trauma center. PATIENTS: Seventeen of the 83 patients identified through trauma, vascular, and/or orthopedic registries consented to participation in this study. INTERVENTIONS: Participating patients completed a study questionnaire, and then underwent a complete vascular examination, including air plethysmographic (APG) assessment. Patients with a history of venous injuries were also studied with color flow duplex venous imaging. MAIN OUTCOME MEASURES: Function of the calf muscle pump as measured by APG, and evidence of chronic venous insufficiency as measured by APG, findings on clinical examination, and by venous ultrasonography. RESULTS: Seventeen patients completed the study, including 8 with a history of vascular injuries, 6 with old fractures, and 3 who had undergone fasciotomy for soft tissue infections. The time from injury to examination ranged from 5 months to 20 years. Eight patients had signs or symptoms of venous insufficiency, the severity of which appeared to be time dependent. The APG data showed significant mean differences between fasciotomy and control extremities in ejection fraction (P<.001) and residual volume fraction (P<.001), both measures of calf muscle pump function. There were no significant changes in venous filling index, a measure of venous reflux, or in outflow fraction, which correlates with venous obstruction. There were no differences in APG variables between patients with vascular injuries vs those with orthopedic or soft tissue injuries. CONCLUSIONS: Lower extremity fasciotomy impairs long-term calf muscle pump function, as measured by APG, in patients with and without vascular injuries. These patients are at risk for the long-term development of chronic venous insufficiency following lower extremity trauma. PMID- 9865657 TI - Postsplenectomy Capnocytophaga canimorsus sepsis presenting as an acute abdomen. AB - Patients with intra-abdominal processes that require prompt surgical intervention, including appendicitis, perforated viscus, ischemic bowel, volvulus, and bowel obstruction, often present with signs and symptoms of an acute abdomen. Several medical problems can mimic an acute abdomen. Overwhelming postsplenectomy infection is a life-threatening condition that can present with acute abdominal symptoms. The incidence of overwhelming postsplenectomy infection ranges from 1% to 25%, and is caused by Streptococcus pneumoniae in 50% of cases. Capnocytophaga canimorsus, a bacteria commonly found in dog saliva, accounts for less than 1% of cases. Overwhelming postsplenectomy infection has a rapidly deteriorating course that progresses to respiratory and renal failure, cardiovascular collapse, and death. The mortality associated with overwhelming postsplenectomy infection is 60% to 80%. Early diagnosis and institution of appropriate antibiotic therapy and supportive care is essential to improve patient outcome. A previously healthy woman who had undergone splenectomy secondary to trauma 11 years earlier presented with symptoms of an acute abdomen. A diagnosis of overwhelming postsplenectomy infection due to C canimorsus was made based on her peripheral blood smear and blood culture findings. Early aggressive care and antibiotic treatment resulted in a successful outcome for this patient with no long-term morbidity. This patient's clinical course demonstrates the importance of early diagnosis and treatment of overwhelming postsplenectomy infection. PMID- 9865658 TI - International medical graduates. PMID- 9865659 TI - Postoperative complications of temporary abdominal surgery. PMID- 9865660 TI - Anatomical studies in antebellum America. PMID- 9865661 TI - Genetic and molecular markers for vascular disease. Overview. PMID- 9865662 TI - Molecular and genetic approaches to the study of cardiovascular disease. PMID- 9865663 TI - Genetic markers: genes involved in atherosclerosis. PMID- 9865664 TI - Genetic markers: genes involved in human hypertension. PMID- 9865665 TI - Genetic markers: genes involved in thrombosis. AB - This article summarizes the genetic markers of human venous and arterial thrombotic disorders. For venous thromboembolism, a factor V mutation (Arg 506- >Gln) has the highest risk, followed by protein C, S and antithrombin III gene defects. By contrast, these genetic defects are not associated significantly with arterial atherothrombotic disorders. Instead, a glycoprotein IIIa polymorphism (Pro33 versus Leu 33) has been reported to be associated with myocardial infarction. Fibrinogen Bbeta chain, factor VII, and plasminogen activator inhibitor-1 gene polymorphisms have been reported to influence the plasma levels of these factors and may indirectly be risk factors for arterial thrombotic disorders. Further studies will uncover additional genetic markers for thrombosis. PMID- 9865666 TI - Molecular markers for atherosclerosis. PMID- 9865667 TI - Cardiovascular risk factors for early carotid atherosclerosis in the general population: the Edinburgh Artery Study. AB - BACKGROUND: Recent attempts to identify cardiovascular risk factors affecting early-stage carotid atherosclerosis, measured by ultrasonographically assessed intima-media thickness, have been inconclusive. OBJECTIVE: To study the relationship between traditional cardiovascular risk factors and intima-media thickness. METHODS: Ultrasonic evaluation of the intima-media thickness of the common carotid artery was included in the 5-year follow-up examination of participants of the Edinburgh Artery Study. We had valid readings of intima-media thickness for 1106 men and women aged 60-80 years. Information on a range of cardiovascular risk factors had been collected during the baseline examination. RESULTS: For men, in addition to age, lifetime smoking (measured in terms of pack years) was the only cardiovascular risk factor associated with increased intima media thickness (P< or = 0.01) in the univariate analysis. Both systolic blood pressure (P < or = 0.001) and the high-density lipoprotein (HDL: total cholesterol ratio (P < or = 0.01) were correlated with intima-media thickness for women. When all the variables had been included in a multivariate analysis, pack years of smoking and the HDL:total cholesterol ratio were associated with early atherosclerotic development in men. In an equivalent analysis for women, alcohol consumption, systolic blood pressure and the HDL:total cholesterol ratio were associated with intima-media thickness. CONCLUSION: These data suggest that risk factors affecting intima-media thickness differ for men and women. Further sex specific analyses of prospective population studies are required in order to clarify the role of 'traditional' cardiovascular risk factors in the early stages of carotid atherosclerosis. PMID- 9865668 TI - Testosterone, sex hormone-binding globulin, lipoproteins, and vascular disease risk. AB - BACKGROUND: That men have a higher risk of coronary heart disease has implicated testosterone as a risk factor. Lipoprotein levels have been reported to be altered by androgens, thus increasing the risk of coronary heart disease, myocardial infarction, and sudden death. The increasing abuse of anabolic steroids and reports of cases of sudden death and myocardial infarction among bodybuilders have raised concerns about an increase in cardiovascular risk for this population. METHODS: Twelve competitive bodybuilders were recruited for a comprehensive study on the cardiovascular risks associated with use of anabolic steroids. Six competitive heavyweight bodybuilders ingesting self-directed regimens of anabolic steroids (steroid group) and six competitive heavyweight drug-free bodybuilders were used for cardiovascular risk assessment. Apolipoproteins A-I and B, total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglyceride, testosterone, and sex hormone-binding globulin levels were determined in each athlete. RESULTS: Total, HDL and LDL cholesterol, apolipoproteins A-I and B, and triglyceride levels were significantly lower in members of the steroid group. As expected, testosterone level was significantly higher in members of the steroid group; sex hormone-binding globulin level was significantly lower. Apolipoprotein and lipoprotein levels were lower in members of the steroid group, whereas the total: HDL cholesterol ratio was significantly higher for members of the steroid group. CONCLUSIONS: Consistently with previous reports, androgens were associated with decreases in HDL cholesterol and apolipoprotein A-I levels. However, androgens were also associated with reduced LDL cholesterol and apolipoprotein B levels. Despite the significantly higher total: HDL cholesterol ratio, the low serum total cholesterol levels (within the fifth percentile) and low plasma triglyceride levels in members of the steroid group raise questions concerning the exact role of androgens in increasing risk of cardiovascular disease. PMID- 9865669 TI - Urinary excretion of nitric oxide metabolites in runners, sedentary individuals and patients with coronary artery disease: effects of 42 km marathon, 15 km race and a cardiac rehabilitation program. AB - BACKGROUND: The mechanisms by which regular exercise is associated with decreases in all-cause and cardiovascular mortality are unknown. Nitric oxide (NO) may have a role, as it is known to be an important factor in cardiovascular regulation. The relationships between physical activity and systemic formation of NO were evaluated in healthy volunteers and in patients with coronary artery disease (CAD). METHODS: Urinary excretion of NO metabolites (nitrates + nitrites) was measured in 50 men. Group 1 comprised 14 highly trained runners (90 km/week) who were tested before and after a marathon race of 42.2 km. Group 2 comprised 11 well trained men (64 km/week) who were tested before and after a 15 km race. Group 3 comprised 12 sedentary individuals who gave a single urine sample. Group 4 comprised 13 patients with CAD who were tested before and after a 6 km walk. RESULTS: Group 1 showed the highest basal levels of urinary NO metabolites: 10.10 mmol/g creatinine; they were followed by group 2, with 5.60 mmol/g creatinine, group 3 with 1.59 mmol/g creatinine and patients with CAD (group 4), who had 0.35 mmol/g creatinine. After the marathon, those in group 1 showed a significant (P=0.0001) reduction of 80% in the excretion of NO metabolites. The 15 km race (group 2 and the 6 km walk (group 4), produced nonsignificant reductions in NO excretion. Patients with CAD were prospectively evaluated before and after a 12 week cardiac rehabilitation program. Their urinary excretion of NO metabolites (mmol/g creatinine) at the end of the program was 157% higher than at baseline (P=0.034). A positive, significant correlation (P=0.006) was observed between the increases in exercise capacity [in METs (one MET is equal to the body's oxygen consumption at rest, and corresponds to 3.5 ml/Kg/min)] and in NO metabolite excretion induced by the 12-week program. CONCLUSIONS: The baseline urinary excretion of NO metabolites increases with increasing levels of physical activity (chronic aerobic exercise). Patients with CAD had lowest levels of urinary NO metabolites and these increased in direct proportion with the gain in functional capacity. These findings suggest that increased NO production may be a major adaptive mechanism by which chronic aerobic exercise training benefits the cardiovascular system. The marked increase in NO production induced by long-term, high levels of aerobic exercise may be protective in athletes undertaking strenuous levels of exercise. PMID- 9865670 TI - International comparison of awareness and attitudes towards coronary risk factor reduction: the HELP study. Heart European Leaders Panel. AB - BACKGROUND: An international survey was conducted to assess public awareness and attitudes to coronary heart disease and to establish the frequency with which certain health-related behaviours are practised in five European countries. METHODS: Members of the general public (n=5013), individuals at increased risk of coronary disease (n=2500), patients who had suffered a myocardial infarction (n=1256) and members of their families (n=1249) were interviewed in a study conducted in France, Germany, Italy, Sweden and the UK. Questions were asked about respondents' attitudes to their health and about their current health practices. RESULTS: The survey revealed a considerable degree of indifference to coronary heart disease, despite the possession of a reasonable level of knowledge of the risks involved, even among patients who had suffered a myocardial infarction. At the same time, respondents declared themselves satisfied with the quality of advice about coronary health that they obtained from the medical profession and regarded these sources of information as highly credible. Media health campaigns, by contrast, had comparatively little impact. CONCLUSION: A survey of five European countries shows that individuals possess reasonable levels of knowledge about coronary heart disease. They also have access to sources of heart health information that are perceived as highly credible. Nonetheless, such information has a very limited impact on their practice of health-related behaviours. PMID- 9865671 TI - Safety and efficacy of Omacor in severe hypertriglyceridemia. AB - BACKGROUND: Severe hypertriglyceridemia is a risk factor for acute pancreatitis, therefore decreasing serum triglyceride concentrations is an important component of risk management. Omega-3 fatty acids are well known hypotriglyceridemic agents, but their efficacy in severe forms of the disorder is not well documented. Our objective was to examine the effects of Omacor, a drug composed of 85% omega-3 fatty acid ethyl esters. METHODS: Forty-two patients with triglyceride concentrations between 5.65 and 22.60 mmol/l (500 and 2000 mg/dl) were studied in a prospective, double-blind, placebo-controlled trial of Omacor (4 g/day for 4 months). RESULTS: Compared with baseline values, Omacor significantly reduced mean triglyceride concentrations by 45% (P<0.00001), cholesterol by 15% (P< 0.001), very-low-density lipoprotein cholesterol by 32% (P< 0.0001) and cholesterol:high density lipoprotein (HDL) cholesterol ratio by 20% (P=0.0013), and increased HDL cholesterol by 13% (P=0.014) and low-density lipoprotein cholesterol by 31% (P=0.0014). The placebo had no effect on these parameters. Omacor was well tolerated and no patient discontinued medication because of side effects. CONCLUSIONS: Four capsules of Omacor per day markedly decreased triglyceride concentrations in patients with severe hypertriglyceridemia. The availability of a potent and safe omega-3 fatty acid preparation for this patient population should diminish the risk for acute pancreatitis, and may also reduce the long-term risk for cardiovascular disease. PMID- 9865672 TI - Which lifestyle parameters discriminate high- from low-risk participants for coronary heart disease risk factors. Longitudinal analysis covering adolescence and young adulthood. AB - BACKGROUND: The purpose of this study was to analyse which lifestyle parameters (daily physical activity, dietary intake, smoking habits and alcohol consumption) discriminate between participants at high-risk and those at low-risk for coronary heart disease (CHD) [systolic and diastolic blood pressure (SBP/DBP), total serum cholesterol (TC), high-density lipoprotein cholesterol (HDL), the TC: HDL ratio, body fatness (sum of skinfolds [SSF]) and cardiopulmonary fitness (VO2-max)]. METHODS: The data were derived from the Amsterdam Growth and Health Study (AGHS); an observational longitudinal study in which six repeated measurements were carried out over a period of 15 years on 181 participants aged 13 years at entry to the study. To assess possible discriminating factors, generalized estimating equations were used. This method makes use of risk group changing behaviour over time by using all available longitudinal data. RESULTS: For DBP no significant relationships were found; high risk for SBP was inversely related to smoking habits (OR=0.52; P<0.01). No significant relationships were found for TC; high risk for HDL was positively related to the intake of carbohydrates (OR=1.2; P=0.02) and to smoking habits (OR=1.6; P=0.04); high risk for the TC:HDL ratio was positively related to the intake of carbohydrates (OR=1.3; P=0.01). High risk for SSF was positively related to the intake of protein (OR=1.5; P<0.01) and smoking habits (OR=1.8; P=0.01) and inversely related to daily physical activity (OR=0.81; P=0.01). High risk for VO2-max was inversely related to daily physical activity (OR=0.67; P<0.01). CONCLUSIONS: In the relative young and healthy population of the AGHS during adolescence and young adulthood, physical inactivity was the most important lifestyle parameter related to high risk for CHD. PMID- 9865674 TI - Bibliography. Current world literature. PMID- 9865675 TI - Cancer vaccines: reborn or just recycled? AB - The field of human cancer vaccines is currently undergoing a revival. This volume describes a variety of approaches to active immunotherapy, some traditional and some more technologically sophisticated. Interpretation of their claims to clinical efficacy requires an understanding of some of the principles of tumor immunotherapy. These include the following: (1) human cancers are weakly immunogenic or not immunogenic at all; (2) cell-mediated immune responses, mainly those of T lymphocytes, are critical to tumor rejection; (3) almost nothing is known about the nature of the rejection antigens on human tumors; (4) it is not clear whether these antigens are different for each case of human cancer or whether there are clinically useful antigens that are shared by all or most tumors of a given histologic type; and (5) the effectiveness of vaccines is greatly limited by excessive tumor burden. The reader is invited to approach this collection of papers with a combination of optimism and skepticism. PMID- 9865673 TI - Genetic variability in the renin-angiotensin system: prevalence of alleles and genotypes. AB - BACKGROUND: Variants of the human genes coding for renin, angiotensin-converting enzyme (ACE), angiotensinogen (AGT) and the angiotensin II type-I receptor (AT1R) are inconsistently associated with cardiovascular-renal disease, possibly because of genetic differences in the background populations. METHODS: This systematic review of the literature investigated genetic variation in the renin system according to race, sex and age. Across studies with relevant information, multivariate analyses also accounted for the methods of genotyping and the enrollment of subjects as controls, cases or groups studied cross-sectionally. RESULTS: The 185 reviewed reports included 64978 subjects. In five studies (n=989) on the renin gene, the Hind III and Taq I polymorphisms varied with the groups' average age, whereas the Bg I, Bg II and Hind III but not Taq I sites differed according to race. Among 135 studies (n=44697) on the deletion/insertion (D/I) polymorphism of the ACE gene, the frequency of the D allele was 54.0%. Its prevalence was not related to sex and black race, was 49.9% lower in Asians, 10.0% lower in studies relying on I-specific primers, 4.9% higher for each 25 year increment in the average age of the groups studied, and 16.7% higher in cases than controls. Among 12 studies (n=4952) on the T174-->M variant of the AGT gene, the M174 frequency was 11.0%, did not vary according to sex and enrollment group, was 56.7% lower in blacks and 39.5% lower for each 25-year increase in the groups' mean age. Across 44 studies (n=16713) on the M235-->T substitution in the AGT gene, the T235 prevalence was 51.6%. Its frequency was not related to sex and the method of genotyping, tended to be 7.5% lower for each 25-year increase in average age, was from 4.6 to 6.6 times higher in nonwhites than whites and 13.2% higher in cases than controls. Among 13 studies (n=4332) on the A1166-->C variant of the AT1R gene, the C1166 allelic frequency was 25.7%. Its prevalence was independent of the enrollment group, 77.4% lower in Asians, and nearly doubled for each 25-year increment in age. CONCLUSION: With adjustments applied for the subjects' enrollment group and the methods of genotyping, genetic polymorphism in the renin system varies according to race and age, but not sex. One possible application of the present results is to provide allelic and genotypic frequencies, which could be used to assess power, to perform sample size calculations, or to predict selection bias in future studies. PMID- 9865676 TI - Active immunotherapy with allogeneic tumor cell vaccines: present status. AB - This review will concentrate on allogeneic vaccines for melanoma The important principles of melanoma vaccine effectiveness are discussed in detail, followed by a review of the progress of several clinical trials investigating allogeneic vaccines. No therapeutic cancer vaccine has yet been approved for general use by the US Food and Drug Administration. However, much progress has been made in the field of vaccine immunotherapy, especially for the treatment of melanoma. Active immunotherapy with tumor vaccines is progressing rapidly as an emerging option for cancer therapy. PMID- 9865677 TI - Perspective on allogeneic melanoma lysates in active specific immunotherapy. AB - We have tested allogeneic melanoma cell lysates as active immunotherapy, originally in stage IV patients to determine their safety and immunologic effectiveness. Surprisingly, phase I and II trials with frozen lysates showed a 20% objective response rate, with 8% long-term survivors. Melacine (Ribi ImmunoChem Research, Hamilton, MT), a lyophilized preparation from the same two cell lines, has been tested nationally and has caused regressions in approximately 10% of patients. Long-term stabilization of disease was noted in 10% to 20% of patients in all trials. A multicenter phase III comparison of low dose cyclophosphamide plus Melacine versus four-drug chemotherapy showed no difference in response rates and survival, with fewer and milder side effects due to Melacine. In our single-arm trial in resected stage III disease, the overall survival rate (66-month median follow-up) is 66%, with a median relapse-free survival time of 36 months. Interferon-alfa 2b (IFN-alpha) given to patients failing to respond to Melacine elicited major objective responses in a larger proportion than anticipated with IFN-alpha alone. These results stimulated current multicenter trials in stage IV and resected stage III melanoma of Melacine and IFN-alpha in combination versus IFN-alpha alone. Of scientific note were (I) identification of a new melanoma antigen from a gene (MG50) isolated from one of the immunizing cell lines, and (2) demonstration that a new melanoma arising in 1995 in a long-term survivor was immunologically and genetically distinct from her original 1986 tumor. While it is important to define which epitopes are involved, multiepitopic (polyvalent) mixtures have established the therapeutic effect of melanoma vaccines. PMID- 9865678 TI - Ganglioside vaccines with emphasis on GM2. AB - Gangliosides are neuraminic acid-containing glycosphingolipids that are anchored into the cell membrane lipid bilayer by lipophilic ceramide chains. They are overexpressed on tissues of neuroectodermal origin, and particularly in tumors such as melanomas, sarcomas, neuroblastomas, astrocytomas, and small cell lung cancers. Both active and passive immunotherapy trials have identified gangliosides as uniquely effective targets for antibody mediated melanoma immunotherapy. Induction of antibodies against GM2 by vaccination has correlated with an improved prognosis in American Joint Committee on Cancer (AJCC) stage III melanoma patients and vaccines containing GM2 chemically conjugated to keyhole limpet hemocyanin (KLH; GM2-KLH) plus the immunologic adjuvant QS-21 have proven to be consistently immunogenic. Phase III trials with this vaccine are ongoing in patients with melanoma in the United States, Canada, Europe, Australia, and New Zealand. GD2, fucosylated GMI, and GD3-KLH conjugates plus QS-21 are also consistently immunogenic, inducing IgM and IgG antibodies in the majority of patients. Polyvalent ganglioside-KLH conjugate plus QS-21 vaccines should be available in early 1999 for testing in phase II and III clinical trials. PMID- 9865679 TI - Autologous, hapten-modified vaccine as a treatment for human cancers. AB - We have devised a novel approach to active immunotherapy based on modification of autologous cancer cells with the hapten, dinitrophenyl (DNP). The treatment program consists of multiple intradermal injections of DNP-modified autologous tumor cells mixed with BCG. Administration of DNP-vaccine to patients with metastatic melanoma induces a unique reaction- the development of inflammation in metastatic masses. Histologically, this consists of infiltration of T lymphocytes, most of which are CD8+. These T cells usually produce interferon gamma in situ. Moreover, they represent expansion of T-cell clones with novel T cell receptor (TCR) structures. Occasionally, administration of DNP-vaccine results in regression of measurable metastases. The most common site of regression has been small lung metastases. Administration of DNP-vaccine to patients in the postsurgical adjuvant setting produces a more striking clinical effect. Of 62 patients with clinically evident stage III melanoma who had undergone lymphadenectomy, the 5-year relapse-free survival rate was 45% and the overall survival rate was 58%. These results appear to be better than those obtained with high-dose interferon, although a randomized phase III trial is required to prove that point. A recent phase I study suggests that this therapeutic approach is also applicable to stage III ovarian cancer. There appear to be no insurmountable impediments to applying this approach to much larger numbers of patients or to developing it as a standard cancer treatment. PMID- 9865680 TI - Heat-shock protein-based anticancer immunotherapy: an idea whose time has come. AB - The heat-shock proteins (HSPs) belong to a family of ubiquitous and abundant proteins used successfully in prophylactic and therapeutic vaccination against tumors. The HSPs are natural chaperones of peptides that reflect the immunologic composition of the cell. Immunization with tumor-derived HSP-peptide complexes (HSP-PC) induces a potent antigen-specific immune response against both the primary tumor and preestablished metastases; the response is restricted to the tumor from which the HSPs are purified. This article describes the manufacturing process of an individual HSP vaccine, and discusses the rationale, feasibility, advantages, and safety of this new approach of cancer immunotherapy in the context of various animal models and of the first HSP clinical trial. PMID- 9865681 TI - Ex-vivo gene therapy using cytokine-transduced tumor vaccines: molecular and clinical pharmacology. AB - Whether the current generation of cytokine gene-transduced tumor vaccines will show clinical efficacy is under study. Fortunately, the large safety profile so far observed with gene-transduced tumor vaccines can allow outpatient testing in large populations of patients in the adjuvant therapy situation. This will allow large studies statistically powered to see potentially important adjuvant therapy effects in the range that are observed for tamoxifen in breast cancer. For example, the outpatient, adjuvant therapy safety context has been established in the use of GM-CSF gene-transduced autologous prostate cancer vaccines following radical prostatectomy. Similar adjuvant therapy clinical trial efforts are anticipated with allogeneic breast, colon, pancreatic, and ovarian cancer in addition to prostate, renal cell carcinoma, and melanoma. The reverse translation of early clinical data back to basic laboratory research also suggests the field of cytokine gene-transduced tumor vaccine research will remain vibrant. Efforts are currently being directed on optimizing DC activation with polycistronic constructs of cytokine genes, and overexpressing the most relevant tumor associated peptides. As in the case of antineoplastic drug development, not all lead compounds will become approved drugs in medical oncology. Rigorous yet innovative clinical trial designs will be key to the accelerated identification of cytokine gene-transduced vaccines that improve survival in cancer patients. PMID- 9865682 TI - Immunization with virus-modified tumor cells. AB - Direct infection of tumor cells with viruses transfering protective or therapeutic genes-a frequently used procedure for production of tumor vaccines in human gene therapy-is often limited by the number of tumor cells that can reliably be infected, as well as by issues of selectivity and safety. In this review, we describe an efficient, selective, and safe way of infecting human tumor cells with a natural virus with interesting pleiotropic immune stimulatory properties, the avian paramyxovirus Newcastle disease virus (NDV). Advantages of this virus are its good cell-binding properties, its selective replication in tumor cell cytoplasm, which is independent of cell proliferation, and its relative safety. Most important for its use as an adjuvant in human cancer vaccine are its ability to introduce T-cell costimulatory activity, to prevent anergy induction, and to induce locally chemokines (eg, RANTES, IP-10) and cytokines (eg, interferon alpha, beta [IFN-alpha, beta] and tumor necrosis factor alpha [TNFalpha]) that affect T-cell recruitment and activation. A further development consists of attachment-via NDV-derived hemagluttinin-neuraminidase (HN) membrane-anchoring molecules-of universal defined bispecific reagents such as T-cell-activating anti-CD28 antibodies. Finally, we summarize the status of our clinical studies with the autologous virus modified live cell vaccine (ATV) NDV. PMID- 9865683 TI - Active immunization against cancer cells: impediments and advances. AB - Immunotherapy of cancer is still mainly an experimental treatment. Some monoclonal antibodies have been approved for adjuvant therapy of cancer in patients, but active immunization strategies have not yet matured to this stage. The fact that vaccination against viral diseases is effective has primed high expectations for successful vaccination against cancer as well. Indeed, in some animal models, therapeutic results could be obtained against short-term established tumors, which paved the way for clinical trials. However, the first results with active immunization in cancer patients were disappointing and this led to a careful examination of current protocols and the search for more effective approaches. Evaluation of the available data suggests that cancer patients may not be comparable in their immune response to cancer vaccines with healthy persons. Furthermore, the tumor seems to be able to develop several immune-escape mechanisms, which either inactivate the specific immune cells or prevent activation of potential effector mechanisms against the tumor. Here, we review the impediments that have been identified in murine models and clinical trials for immunotherapy of cancer. It will be important to study the hurdles to come to a better understanding of the immune evasion of tumors and to achieve efficient activation of the immune system in cancer patients against the tumor. This knowledge will open new possibilities for active immunization against cancer. PMID- 9865684 TI - An expanding role for docetaxel. PMID- 9865685 TI - Phase III studies of single-agent docetaxel in patients with metastatic breast cancer who have progressed despite previous chemotherapy regimens: preliminary results. AB - A recent large phase III trial has for the first time demonstrated that choice of treatment can influence survival duration in patients with metastatic breast cancer who have progressed despite previous anthracycline-containing therapy. In a multicenter study, patients who received docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) experienced a longer median survival time (II.4 months v 8.7 months; P = .0097) as well as a longer time to progression (19 weeks v II weeks; P < .001) and higher overall response rate (30% v II.6%; P < .0001) than patients receiving treatment with mitomycin C and vinblastine. The toxicity profile was manageable and tolerable for both arms. Evidence for a risk to benefit ratio favoring docetaxel is also provided by a second phase III trial in which docetaxel was compared with doxorubicin in metastatic breast cancer patients who have progressed despite prior alkylating chemotherapy. In these patients, docetaxel was more active than doxorubicin, achieving an overall response at a significantly higher rate (47.8% v 33.3%; P = .008) and in a shorter time (median, 12 weeks v 23 weeks; P = .007). In this study, the duration of survival was not influenced by treatment. However, the higher response rate with docetaxel was achieved without the risk of potentially fatal cardiac toxicity seen in some patients who received doxorubicin. To date, docetaxel is the only single agent shown to have a potential superior activity when compared with doxorubicin in patients with progressive metastatic disease. PMID- 9865686 TI - Compilation of phase I and II trial data of docetaxel and doxorubicin in the treatment of advanced breast cancer and other malignancies. AB - Doxorubicin and docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) are the most active cytotoxic agents in the treatment of advanced breast cancer. In the pre-taxane era, randomized trials demonstrated that doxorubicin-containing chemotherapy regimens are associated with higher response rates and improved survival compared with non-doxorubicin-containing regimens. Doxorubicin containing regimens were therefore considered the standard of care at that time. Because of the substantial activity of the taxanes in anthracyline-resistant breast cancer, there is a clear rationale for combining doxorubicin with the taxanes as initial therapy for patients with metastatic disease. The combination of doxorubicin plus paclitaxel has been extensively studied. Impressive response rates were noted in at least two studies, although the treatment also resulted in an unacceptably high risk of congestive heart failure. Paclitaxel is known to perturb the metabolism of doxorubicin, thereby increasing drug exposure and augmenting the cardiotoxicity of the anthracycline. The cardiotoxicity of the combination may be ameliorated by restricting the cumulative doxorubicin dose to less than 360 mg/m2, by increasing the interval between administration of the two drugs to at least 4 hours, or by adding the cardioprotective agent dexrazoxane. The Eastern Cooperative Oncology Group performed a randomized phase III trial comparing doxorubicin alone, paclitaxel alone, or the doxorubicin/paclitaxel combination in patients with metastatic breast cancer. Although the response rate and time to treatment failure improved with the combination, survival did not. There have been two phase II trials investigating the combination of doxorubicin plus docetaxel. The regimen was highly effective and did not seem to be associated with an increased risk of congestive heart failure. These findings justify further evaluation of the doxorubicin/docetaxel combination in patients with advanced and operable breast cancer. Such trials are currently in progress and will define the role for this combination in the management of patients with breast cancer. PMID- 9865687 TI - Taxoids in combination with anthracyclines and other agents: pharmacokinetic considerations. AB - The combination in clinical trials of taxoids with doxorubicin has focused attention on possible drug interactions. One specific finding requiring explanation is the relative lack of cardiotoxicity of the docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France)/doxorubicin combination compared with that of the paclitaxel/doxorubicin combination. Data in mice demonstrate that epirubicin concentrations in cardiac tissue 24 hours after treatment are approximately doubled by the coadministration of paclitaxel. This effect appears to be less marked with docetaxel administered in its normal polysorbate vehicle or when the drug is given in Cremophor EL (Sigma, St Louis, MO). Both Cremophor EL or polysorbate appear to cause an increase in epirubicin tissue levels, although the levels were less than those seen with paclitaxel. Pharmacokinetic data from women being treated with combination therapy for advanced breast cancer demonstrate that the administration of docetaxel following doxorubicin does not alter doxorubicin's area under the plasma concentration-time, curve, maximum plasma concentration, or time until maximum plasma concentration is reached. However, the area under the curve of docetaxel is significantly increased by the prior administration of doxorubicin. These findings may explain both the low cardiotoxicity and the high clinical efficacy of the docetaxel/doxorubicin combination. Phase I clinical trials of combinations in which docetaxel was used together with vinorelbine, ifosfamide, or 5-fluorouracil have shown no evidence of relevant pharmacokinetic interactions. PMID- 9865688 TI - Weekly schedules of docetaxel. AB - The weekly administration of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) at doses up to and including 40 mg/m2 induces low levels of hematologic and nonhematologic toxicity. In particular, weekly scheduling appears to markedly reduce the incidence of neutropenia compared with regimens in which an equivalent dose rate (mg/m2/wk) is given once every 3 weeks. Encouraging responses have been reported in patients with a range of tumor types, including breast cancer, treated with weekly docetaxel. Further trials of weekly docetaxel are in progress, and it appears that this schedule may prove particularly valuable in certain elderly patients who are unsuited to more aggressive chemotherapy. PMID- 9865689 TI - IkappaB-NF-kappaB structures: at the interface of inflammation control. PMID- 9865690 TI - Y do we drink? PMID- 9865691 TI - Smads: transcriptional activators of TGF-beta responses. PMID- 9865692 TI - Spatial control of actin filament assembly: lessons from Listeria. PMID- 9865693 TI - Structure of an IkappaBalpha/NF-kappaB complex. AB - The inhibitory protein, IkappaBalpha, sequesters the transcription factor, NF kappaB, as an inactive complex in the cytoplasm. The structure of the IkappaBalpha ankyrin repeat domain, bound to a partially truncated NF-kappaB heterodimer (p50/ p65), has been determined by X-ray crystallography at 2.7 A resolution. It shows a stack of six IkappaBalpha ankyrin repeats facing the C terminal domains of the NF-kappaB Rel homology regions. Contacts occur in discontinuous patches, suggesting a combinatorial quality for ankyrin repeat specificity. The first two repeats cover an alpha helically ordered segment containing the p65 nuclear localization signal. The position of the sixth ankyrin repeat shows that full-length IkappaBalpha will occlude the NF-kappaB DNA-binding cleft. The orientation of IkappaBalpha in the complex places its N- and C terminal regions in appropriate locations for their known regulatory functions. PMID- 9865694 TI - The crystal structure of the IkappaBalpha/NF-kappaB complex reveals mechanisms of NF-kappaB inactivation. AB - IkappaBalpha regulates the transcription factor NF-kappaB through the formation of stable IkappaBalpha/NF-kappaB complexes. Prior to induction, IkappaBalpha retains NF-kappaB in the cytoplasm until the NF-kappaB activation signal is received. After activation, NF-kappaB is removed from gene promoters through association with nuclear IkappaBalpha, restoring the preinduction state. The 2.3 A crystal structure of IkappaBalpha in complex with the NF-kappaB p50/p65 heterodimer reveals mechanisms of these inhibitory activities. The presence of IkappaBalpha allows large en bloc movement of the NF-kappaB p65 subunit amino terminal domain. This conformational change induces allosteric inhibition of NF kappaB DNA binding. Amino acid residues immediately preceding the nuclear localization signals of both NF-kappaB p50 and p65 subunits are tethered to the IkappaBalpha amino-terminal ankyrin repeats, impeding NF-kappaB from nuclear import machinery recognition. PMID- 9865695 TI - Transmembrane signaling across the ligand-gated FhuA receptor: crystal structures of free and ferrichrome-bound states reveal allosteric changes. AB - FhuA protein facilitates ligand-gated transport of ferrichrome-bound iron across Escherichia coli outer membranes. X-ray analysis at 2.7 A resolution reveals two distinct conformations in the presence and absence of ferrichrome. The monomeric protein consists of a hollow, 22-stranded, antiparallel beta barrel (residues 160 714), which is obstructed by a plug (residues 19-159). The binding site of ferrichrome, an aromatic pocket near the cell surface, undergoes minor changes upon association with the ligand. These are propagated and amplified across the plug, eventually resulting in substantially different protein conformations at the periplasmic face. Our findings reveal the mechanism of signal transmission and suggest how the energy-transducing TonB complex senses ligand binding. PMID- 9865696 TI - SARA, a FYVE domain protein that recruits Smad2 to the TGFbeta receptor. AB - Smads transmit signals from transmembrane ser/thr kinase receptors to the nucleus. We now identify SARA (for Smad anchor for receptor activation), a FYVE domain protein that interacts directly with Smad2 and Smad3. SARA functions to recruit Smad2 to the TGFbeta receptor by controlling the subcellular localization of Smad2 and by interacting with the TGFbeta receptor complex. Phosphorylation of Smad2 induces dissociation from SARA with concomitant formation of Smad2/Smad4 complexes and nuclear translocation. Furthermore, mutations in SARA that cause mislocalization of Smad2 inhibit TGFbeta-dependent transcriptional responses, indicating that the regulation of Smad localization is important for TGFbeta signaling. These results thus define SARA as a component of the TGFbeta pathway that brings the Smad substrate to the receptor. PMID- 9865697 TI - Mammalian Grb2 regulates multiple steps in embryonic development and malignant transformation. AB - Proteins with SH2 and SH3 domains link tyrosine kinases to intracellular pathways. To investigate the biological functions of a mammalian SH2/SH3 adaptor, we have introduced a null mutation into the mouse gene for Grb2. Analysis of mutant embryonic stem cells, embryos, and chimeras reveals that Grb2 is required during embyrogenesis for the differentiation of endodermal cells and formation of the epiblast. Grb2 acts physiologically as an adaptor, since replacing the C terminus of the Ras activator Sos1 with the Grb2 SH2 domain yields a fusion protein that largely rescues the defects caused by the Grb2 mutation. Furthermore, Grb2 is rate limiting for mammary carcinomas induced by polyomavirus middle T antigen. These data provide genetic evidence for a mammalian Grb2-Ras signaling pathway, mediated by SH2/SH3 domain interactions, that has multiple functions in embryogenesis and cancer. PMID- 9865698 TI - Role of WUSCHEL in regulating stem cell fate in the Arabidopsis shoot meristem. AB - The shoot meristem gives rise to the aerial parts of higher plants by continuously initiating new organs. The basis of this activity is its ability to maintain a pool of pluripotent stem cells, which are the ultimate source of all tissues of the shoot. In Arabidopsis plants mutant for the WUSCHEL (WUS) gene, the stem cells are misspecified and appear to undergo differentiation. Here, we show that WUS encodes a novel homeodomain protein which presumably acts as a transcriptional regulator. The pattern of WUS expression suggests that stem cells in the shoot meristem are specified by an underlying cell group which is established in the 16-cell embryo and becomes localized to its prospective domain of function by asymmetric cell divisions. PMID- 9865699 TI - LIM homeodomain factors Lhx3 and Lhx4 assign subtype identities for motor neurons. AB - The circuits that control movement are comprised of discrete subtypes of motor neurons. How motor neuron subclasses develop and extend axons to their correct targets is still poorly understood. We show that LIM homeodomain factors Lhx3 and Lhx4 are expressed transiently in motor neurons whose axons emerge ventrally from the neural tube (v-MN). Motor neurons develop in embryos deficient in both Lhx3 and Lhx4, but v-MN cells switch their subclass identity to become motor neurons that extend axons dorsally from the neural tube (d-MN). Conversely, the misexpression of Lhx3 in dorsal-exiting motor neurons is sufficient to reorient their axonal projections ventrally. Thus, Lhx3 and Lhx4 act in a binary fashion during a brief period in development to specify the trajectory of motor axons from the neural tube. PMID- 9865700 TI - Randomization of left-right asymmetry due to loss of nodal cilia generating leftward flow of extraembryonic fluid in mice lacking KIF3B motor protein. AB - Microtubule-dependent motor, murine KIF3B, was disrupted by gene targeting. The null mutants did not survive beyond midgestation, exhibiting growth retardation, pericardial sac ballooning, and neural tube disorganization. Prominently, the left-right asymmetry was randomized in the heart loop and the direction of embryonic turning. lefty-2 expression was either bilateral or absent. Furthermore, the node lacked monocilia while the basal bodies were present. Immunocytochemistry revealed KIF3B localization in wild-type nodal cilia. Video microscopy showed that these cilia were motile and generated a leftward flow. These data suggest that KIF3B is essential for the left-right determination through intraciliary transportation of materials for ciliogenesis of motile primary cilia that could produce a gradient of putative morphogen along the left right axis in the node. PMID- 9865701 TI - A VSG expression site-associated gene confers resistance to human serum in Trypanosoma rhodesiense. AB - Infectivity of Trypanosoma brucei rhodesiense to humans is due to its resistance to a lytic factor present in human serum. In the ETat 1 strain this character was associated with antigenic variation, since expression of the ETat 1.10 variant surface glycoprotein was required to generate resistant (R) clones. In addition, in this strain transcription of a gene termed SRA was detected in R clones only. We show that the ETat 1.10 expression site is the one selectively transcribed in R variants. This expression site contains SRA as an expression site-associated gene (ESAG) and is characterized by the deletion of several ESAGs. Transfection of SRA into T.b. brucei was sufficient to confer resistance to human serum, identifying this gene as one of those responsible for T.b. rhodesiense adaptation to humans. PMID- 9865702 TI - Fab1p PtdIns(3)P 5-kinase function essential for protein sorting in the multivesicular body. AB - Sorting of signal-transducing cell surface receptors within multivesicular bodies (MVBs) is required for their rapid down-regulation and degradation within lysosomes. Yeast mutants defective in late stages of transport to the vacuole/lysosome accumulate MVBs. We demonstrate that the membrane glycoprotein carboxypeptidase S and the G protein-coupled receptor Ste2p are targeted into the vacuole lumen, and this process requires a subset of VPS gene products essential for normal endosome function. The PtdIns(3)P 5-kinase activity of Fab1p, which converts the product of the Vps34p PtdIns 3-kinase PtdIns(3)P into PtdIns(3,5)P2, also is required for cargo-selective sorting into the vacuole lumen. These findings demonstrate a role for phosphoinositide signaling at distinct stages of vacuolar/lysosomal protein transport and couple PtdIns(3,5)P2 synthesis to regulation of MVB sorting. PMID- 9865703 TI - A role for dystroglycan in basement membrane assembly. AB - Basement membranes are composed of ordered arrays of characteristic extracellular matrix proteins, but little is known about the assembly of these structures in vivo. We have investigated the function of dystroglycan, a cell-surface laminin receptor expressed by cells contacting basement membranes in developing and adult tissues. We find that dystroglycan is required for the formation of a basement membrane in embryoid bodies. Our results further indicate that dystroglycanlaminin interactions are prerequisite for the deposition of other basement membrane proteins. Dystroglycan may exert its influence on basement membrane assembly by binding soluble laminin and organizing it on the cell surface. These data establish a role for dystroglycan in the assembly of basement membranes and suggest fundamental mechanisms underlying this process. PMID- 9865704 TI - Efficacy of paclitaxel or doxorubicin used as single agents in advanced breast cancer: a literature survey. AB - The anthracyclines are among the most active agents in the treatment of breast cancer. In recent years, the taxoids have produced promising results as single agents in breast cancer. Average overall response rates of 43% in previously untreated and 29% in pretreated metastatic breast cancer have been reported in phase II trials with paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ). Based on the known activity of doxorubicin and the single-agent phase II results of paclitaxel, two phase III randomized trials comparing these drugs as monotherapy in first-line, with crossover on progression, were conducted by the European Organization for Research and Treatment of Cancer and the Intergroup. The preliminary results of these studies were presented at international congresses and will be discussed here. PMID- 9865706 TI - Evolution in the treatment of advanced breast cancer. AB - There has been a gradual evolution in the philosophy of treatment for metastatic breast cancer. It has long been known that endocrine therapy, radiotherapy, and chemotherapy could offer substantial palliative benefits to patients with symptomatic metastases. While these quality of life issues remain crucially important, it is increasingly recognized that the survival of patients with this condition also appears to be improving as a result of therapeutic advances. Chemotherapy appears to prolong the median duration of survival of patients, probably by approximately 1 year. It is the common experience of oncologists that patients with life-threatening visceral metastases may be rescued with chemotherapy. Until recently, doxorubicin was considered to be the most active single agent available for the treatment of metastatic breast cancer, with response rates of 40% to 50% commonly reported. Other active agents include cisplatin, cyclophosphamide, 5-fluorouracil, ifosfamide, methotrexate, mitomycin C, mitoxantrone, vinblastine, and vincristine. Anthracycline-containing regimens were found to be more effective than combinations without anthracyclines, but overall, it is not clear whether combinations are superior to high-dose single agent anthracyclines. Since the development of the cyclophosphamide/doxorubicin/5 fluorouracil regimen, no major improvements on combination regimens have been achieved until the discovery of the taxoids. These new agents have substantial activity against metastatic breast cancer. The results of phase II studies suggest that of these agents, used at the recommended doses, docetaxel (Taxotere, Rhone-Poulenc Rorer, Antony, France) may be the most active, achieving an objective response rate of 59% in minimally pretreated patients and 47% when used in second-line treatment. In these studies, docetaxel was given at the standard dose of 100 mg/m2 over 1 hour. Recent results from phase III studies in which individual studies with docetaxel and paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) have been compared with standard therapies indicate that docetaxel is the most active single agent in metastatic breast cancer. PMID- 9865705 TI - Docetaxel versus doxorubicin in patients with metastatic breast cancer who have failed alkylating chemotherapy: a preliminary report of the randomized phase III trial. 303 Study Group. AB - Three hundred twenty-six patients who had failed prior alkylating agents, given either as adjuvant therapy or therapy for advanced breast cancer or both, were randomly assigned to treatment with up to seven cycles of doxorubicin 75 mg/m2 or docetaxel (Taxotere, Rhone-Poulenc Rorer, Antony, France) 100 mg/m2 given every 3 weeks. The two arms of the study were well-matched for age, performance status, previous therapy, and the nature of the metastatic disease. Forty-seven percent of the docetaxel-treated patients and 49% of the doxorubicin-treated patients were defined as having disease that showed primary or secondary resistance. Overall, 48% of patients treated with docetaxel had an objective response. This was significantly greater than the 33% response rate seen in doxorubicin-treated patients (P = .008). Among those patients classified as having resistant disease, the difference was equally clear cut (response rate, 47% v 25%; P = .003). Overall median time to response was 12 weeks with docetaxel and 23 weeks with doxorubicin. Febrile neutropenia, grade 3/4 nausea and vomiting, and cardiotoxicity were significantly more common among the doxorubicin-treated patients, while diarrhea grade 3-4, skin toxicity, neurologic toxicity, fluid retention, and allergy of any grade were significantly more likely in the docetaxel-treated patients. This study demonstrates for the first time the superiority in terms of response rate of a taxoid over an anthracycline in the treatment of advanced breast cancer. PMID- 9865707 TI - Taxanes in combination with doxorubicin in the treatment of metastatic breast cancer. AB - Given their high level of activity when used as single agents in metastatic breast cancer, the combination of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) with doxorubicin is a logical development in the search for regimens that will improve prognosis in this disease. In a phase I trial conducted at the Institut Curie and Hospital Paul-Brousse (Paris, France), 42 women with previously untreated metastatic breast cancer received a total of 286 cycles of the combination at six dose levels. Prophylactic steroids and H1 and H2 blockers were given. The maximum tolerated dose was 85 mg/m2 docetaxel plus 50 mg/m2 doxorubicin. Except for neutropenia, no grade 3-4 or severe nonhematologic toxicities were seen. The incidence of fluid retention was low: moderate toxicity in only 19% of patients and no severe cases. No cases of congestive heart failure or symptomatic decrease in left ventricular ejection fraction occurred (median cumulative doxorubicin dose, 392 mg/m2; range, 240 to 559 mg/m2). High activity was observed at all dose levels. For patients receiving one of the four highest dose levels, the overall response rate was 81% (95% confidence intervals, 63% to 93%); comparable levels of response were seen at all disease sites. The doses recommended for phase II/III studies of this active and well-tolerated combination are either 50 mg/m2 doxorubicin plus 75 mg/m2 docetaxel or 60 mg/m2 of both drugs. Encouraging response rates also have been seen in studies in which paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) was combined with doxorubicin, although cardiotoxicity was a significant factor in some studies. PMID- 9865708 TI - Taxoids in combination with epirubicin: the search for improved outcomes in breast cancer. AB - The combination of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) and either doxorubicin or epirubicin has significant activity against metastatic breast cancer. However, the optimal schedule in terms of activity and toxicity is still under investigation. The use of docetaxel (Taxotere; Rhone Poulenc Rorer, Antony, France) in combination with anthracyclines is also active and could represent a safe and favorable alternative. In a phase I trial, 28 patients with metastatic or locally advanced disease received combination chemotherapy using doses of up to 85 mg/m2 docetaxel and 120 mg/m2 epirubicin, with granulocyte colony-stimulating factor support at the higher dose levels. Neutropenia was the dose-limiting toxicity. Severe fluid retention and symptomatic cardiotoxicity were not seen. Responses occurred at doses of 75 mg/m2 docetaxel plus 90 mg/m2 epirubicin and above. However, the proportion of responders did not increase with increasing dose, and the 75 mg/m2 docetaxel/90 mg/m2 epirubicin combination has been selected for further phase II study. PMID- 9865709 TI - Taxane-based three-drug combination in metastatic and adjuvant treatment of breast cancer. AB - Considering the recommended dose of the docetaxel/doxorubicin combination (75 mg/m2 and 50 mg/m2, respectively), we decided to proceed with a pilot program in untreated metastatic breast cancer aimed at defining a multidrug regimen that could be later randomly compared with a standard doxorubicin-containing polychemotherapy regimen with equidoses of doxorubicin such as the FAC protocol (5-fluorouracil 500 mg/m2, doxorubicin 50 mg/m2, and cyclophosphamide 500 mg/m2) in first-line metastatic and adjuvant treatment of breast cancer patients. We proceeded with a pilot phase II study of the TAC combination, which consists of docetaxel 75 mg/m2 as a 1-hour infusion preceded by doxorubicin 50 mg/m2 and cyclophosphamide 500 mg/m2, both given as an intravenous bolus. Three hundred seventy courses were delivered in 54 anthracycline-naive patients, among whom 62% had visceral metastases. Median relative dose intensity was above 98% for all drugs. Grade 4 neutropenia was the main toxicity (70% of cycles) and the incidence of febrile neutropenia and infection was acceptable (6% and 0.8% of cycles, respectively). Acute and chronic extrahematologic toxicities were mild, mostly grade 2, and the docetaxel-specific toxicities (fluid retention, nail changes, etc) were not major clinical problems; no patient was discontinued due to fluid retention. The major response rate was 73% overall and 79% in measurable disease. Time to progression and survival are still under evaluation. The TAC combination is an active and well-tolerated regimen that is the basis of two currently open, pivotal, randomized phase III trials comparing TAC with FAC in the metastatic and adjuvant treatment of breast cancer. PMID- 9865710 TI - Is there a place for "dose-dense" weekly schedules of the taxoids? AB - The recommended weekly dose and the maximum tolerated weekly dose of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) have yet to be determined. We report that a weekly dose of up to 40 mg/m2 docetaxel for 6 weeks is active in pretreated patients with metastatic breast cancer. From a preliminary study, this dose-dense schedule appears to induce less hematologic toxicity than a schedule of 100 mg/m2 every 3 weeks while achieving similar response rates and may represent a valuable alternative involving a shorter treatment time in the palliative therapy of advanced disease in higher-risk patients. The dose-dense weekly administration of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) also appears to be active, although the toxicity profiles of the two taxanes may differ. PMID- 9865711 TI - Cancer as a complex developmental disorder--nineteenth Cornelius P. Rhoads Memorial Award Lecture. AB - The processes of differentiation and tumorigenesis have been long thought to be connected. The recent identification of Patched, a gene essential for Drosophila embryonic development, as a tumor suppressor has focused attention on the concept that tumorigenesis involves abnormalities of development. In fact, a large number of genes in the signalling pathway of the Patched gene are either tumor suppressors or oncogenes. This supports the concept that growth control is a critical requirement of differentiation, and that aberrant cellular development can contribute to malignancy. Whereas the identification of genes that result in dominantly inherited cancer syndromes has played a vital role in understanding cancer, the vast majority of "sporadic" cancers have properties of a complex genetic disease. Approaches to identify common alleles in cancer-associated genes promise to increase our understanding of the disease and aid the rational design of preventative and therapeutic strategies. PMID- 9865712 TI - Loss of atm radiosensitizes multiple p53 null tissues. AB - An unusual clinical finding in ataxia-telangiectasia, a human disorder caused by mutations in atm, is exquisite sensitivity to gamma irradiation. By contrast, homozygous deletion of p53 is marked by radiation resistance in certain tissue compartments. Previous studies (A. J. Levine, Cell, 88: 323-331, 1997) have shown that, in vitro, p53-deficient bone marrow cells are resistant to gamma irradiation. Furthermore, the gastrointestinal radiosensitization engendered by the loss of atm has recently been shown (C. H. Westphal et al., Nat. Genet., 16: 397-401, 1997) to be independent of p53. Expanding on previous work, we have looked at in vivo bone marrow resistance in p53-deficient mice. Our results indicate that inbred FVB strain p53 null mice survive lethal irradiation doses because of bone marrow resistance. Moreover, the deletion of atm radiosensitizes even p53 null bone marrow and mouse embryonic fibroblast cells. The results presented here argue that the loss of atm radiosensitizes multiple tissues in a p53-independent manner. Hence, functional inhibition of atm in p53 null and p53 wild-type human tumors may be a useful adjunct to gamma irradiation-based antitumor therapy. PMID- 9865713 TI - High incidence of translocations t(11;14)(q13;q32) and t(4;14)(p16;q32) in patients with plasma cell malignancies. AB - Abnormalities involving the 14q32 region are recurrent chromosomal changes in plasma cell malignancies. Recent preliminary molecular analyses found IGH rearrangements in almost 100% of human myeloma cell lines and in 75% of patients. However, no systematic study analyzing the nature of the partner chromosomal regions have been reported thus far. To define the exact incidence of illegitimate IGH rearrangements and the respective incidence of partner genes cloned to date, we analyzed 141 patients with either multiple myeloma (MM, n = 127) or primary plasma cell leukemia (PCL, n = 14) using fluorescence in situ hybridization. The overall incidence of illegitimate recombinations was 57% (80 of 141 patients). Analysis of this incidence according to Durie and Salmon stage, patients' status, i.e., MM versus primary PCL and diagnosis versus relapse, immunoglobulin type and subtype, and beta2-microglobulin value, did not show any correlation. To analyze the nature of the partner chromosomal region, we selected probes specific for the following genes: FGFR3 (4p16), MYC (8q24), CCND1 (11q13), MAF (16q23), and BCL2 (18q21). These probes, combined with differentially labeled 14q32 probes, were used for dual-color fluorescence in situ hybridization on interphase plasma cells. Among the 80 patients with illegitimate IGH rearrangement, we identified 23 IGH-CCND1 fusion cases [i.e., t(11;14)], 17 IGH FGFR3 fusion cases [i.e., t(4;14)], 3 IGH-MYC fusion cases [i.e., t(8;14)], and only one IGH-MAF fusion case. No IGH-BCL2 fusion case was detected. In 37 of 80 patients, none of these partner genes was involved. Analysis of cases with specific translocations according to their bioclinical features at diagnosis did not show any correlation. This study demonstrated that CCND1 and FGFR3 genes are involved together in about 50% of MM and primary PCL patients with illegitimate IGH rearrangements. PMID- 9865714 TI - Activation of inducible nitric oxide synthase results in nitric oxide-mediated radiosensitization of hypoxic EMT-6 tumor cells. AB - EMT-6 cells treated for 16 h with 1-10 units/ml IFN-gamma showed a gradual activation of inducible nitric oxide synthase (iNOS) in Western and Northern blots, a simultaneous raise in NO output, and an increase in hypoxic cell radiosensitivity almost to the level of aerobic cells. Both the NO signal and radiosensitization were counteracted by the NO scavenger oxyhemoglobin, by the specific iNOS inhibitor aminoguanidine, and by the L-arginine analogue N(G) monomethyl-L-arginine. Collectively, these data demonstrate that IFN-gamma can radiosensitize EMT-6 cells through iNOS induction and that NO is the effector molecule responsible for radiosensitization. Compared with the spontaneous NO releaser (2)-1-[N-(3-ammoniopropyl)-N-(n-propyl)amino)diazen-1-ium -1,2-diolate], the iNOS-generated NO signal appeared to be 10 times lower yet resulting in the same enhancement ratio of 2.4. Direct stimulation of NO synthesis in tumor cells through the L-arginine/iNOS pathway represents a novel approach to exploit the radiosensitizing properties of NO. PMID- 9865715 TI - Homologous recombination between p53 and its pseudogene in a radiation-induced mouse tumor. AB - Genome recombination is essential for life; however, its dysfunction causes cancer. Here we report the formation of a chimera structure of the p53 gene due to homologous recombination with the p53 pseudogene in tumors produced by repeated local beta-irradiation of the backs of mice. The recombination occurred near the 5' end of exon 5. Because this tumor carried a 5-bp deletion in exon 6 of the expressed p53 allele, and the defect in p53 is reported to elevate the cellular recombination activity, this chimera formation is thought to be initiated by a radiation-induced DNA double strand break in the p53-mutated cell with enhanced recombination. The abundance of this chimera structure was estimated to be 8% of the total of tumor p53, and the functional p53 side of this chimera had no deletion in exon 6. The indication is that the recombination occurred before the loss of heterozygosity of the mutated p53 allele took place but after a few divisions of the original heterozygous p53-mutated cell toward monoclonal expansion. A novel mechanism of cancer induction is suggested. PMID- 9865716 TI - Identification of 12p as a region of frequent deletion in advanced prostate cancer. AB - The identification of homozygous deletions in malignant tissue has been a powerful tool for the localization of tumor suppressor genes. Representational difference analysis (RDA) uses selective hybridization and the PCR to isolate regions of chromosomal loss and has facilitated the identification of tumor suppressor genes such as BRCA2 and PTEN. Twenty RDA clones were generated by comparing genomic DNA from a prostate cancer xenograft to the same patient's normal kidney DNA. Southern blot analysis of the tester and driver and of normal and xenograft DNA, using the differential products as probes, showed the homozygous deletion in 16 of 20 RDA clones. The sequence of one of the differential products overlapped HSU59962, a genomic GenBank sequence on chromosome 12p12-13. Multiplex PCR of the xenograft DNA using polymorphic repeats mapped the deletion to a 1-5-cM region on 12p. Genomic DNA isolated from a panel of cryostat microdissected metastatic prostate adenocarcinomas/normal pairs was screened for loss of heterozygosity using the same polymorphic repeats. Loss of heterozygosity was demonstrated in 9 (47%) of 19 patients. This region may contain, or lie in close proximity to, tumor suppressor genes important in the progression and/or initiation of prostate cancer. PMID- 9865717 TI - Overexpression of the tumor suppressor gene Smad4/DPC4 induces p21waf1 expression and growth inhibition in human carcinoma cells. AB - The Smad4/DPC4 protein functions as a key transcription factor in transforming growth factor beta (TGF-beta) signaling pathways. However, the downstream target genes regulated by Smad4/DPC4 have not been identified until now. We previously demonstrated that the loss of TGF-beta-induced p21waf1 expression and growth inhibition correlates with inactivation of the Smad4/DPC4 gene. Now we show that transient overexpression of Smad4/DPC4 can induce p21waf1 expression, specific Smad4 DNA binding activity, SBE4-luc reporter gene activity, and subsequent growth inhibition in Smad4/DPC4-null cells and other carcinoma cells in the presence or absence of TGF-beta. Taken together, these data show that p21waf1 is one of the Smad4/DPC4-regulated downstream target genes and suggest that overexpression of the Smad4/DPC4 gene can bypass TGF-beta receptor activation and reestablish one of the key regulatory controls of cell proliferation. PMID- 9865718 TI - Differential mucin MUC7 gene expression in invasive bladder carcinoma in contrast to uniform MUC1 and MUC2 gene expression in both normal urothelium and bladder carcinoma. AB - Mucins (MUCs) are high molecular weight membrane glycoproteins. The gene expression of MUCs (MUC1-MUC8) may change characteristically during malignant transformation of epithelial tissues. Total RNA was isolated from the four bladder cancer cell lines RT4, 647V, HT1376, and 486P (pathological gradings between G1 and G4) and 17 samples of transitional cell carcinomas, as well as 16 samples of normal human urothelium of the bladder from surgically removed specimens. The RNA samples were studied with MUC1-, MUC2- and MUC7-specific nested reverse transcription-PCRs. Gene expression of MUC1 and MUC2 was found positive in all normal, as well as in malignant, tissue samples and in the tumor cell lines. In contrast, gene expression of MUC7 was only detected in bladder cancer cell lines and samples of invasive transitional cell carcinomas, but neither in superficial, noninvasive bladder tumors nor normal bladder urothelium. Only one of the samples of normal urothelium obtained from 16 different tumor bearing bladders was positive for MUC7 gene expression. These results suggest a differential MUC7 gene expression with the onset of malignant transformation of the bladder urothelium. PMID- 9865719 TI - The PTEN/MMAC1 tumor suppressor induces cell death that is rescued by the AKT/protein kinase B oncogene. AB - PTEN/MMAC1 is a tumor suppressor gene that is mutated in a variety of cancers. PTEN encodes a phosphatase that recognizes phosphoprotein substrates and the phospholipid, phosphatidylinositol-3,4,5-triphosphate. PTEN inhibited cell growth and/or colony formation in all of the epithelial lines tested with one exception. The decrease in cellular proliferation was associated with an induction of apoptosis and an inhibition of signaling through the phosphatidylinositol 3' kinase pathway. Akt/protein kinase B, a gene whose antiapoptotic function is regulated by phosphatidylinositol-3,4,5-triphosphate, was able to rescue cells from PTEN-dependent death. PTEN, therefore, appears to suppress tumor growth by regulating phosphatidylinositol 3'-kinase signaling. PMID- 9865720 TI - Adeno-associated virus-mediated delivery of antiangiogenic factors as an antitumor strategy. AB - Antiangiogenic tumor therapies have recently attracted intense interest for their broad-spectrum action, low toxicity, and, in the case of direct endothelial targeting, an absence of drug resistance. To promote tumor regression and to maintain dormancy, antiangiogenic agents need to be chronically administered. Gene therapy offers a potential way to achieve sustained therapeutic release of potent antiangiogenic substances. As a step toward this goal, we have generated recombinant adeno-associated virus (rAAV) vectors that carry genes coding for angiostatin, endostatin, and an antisense mRNA species against vascular endothelial growth factor (VEGF). These rAAVs efficiently transduced three human tumor cell lines tested. Transduction with an rAAV-encoding antisense VEGF mRNA inhibited the production of endogenous tumor cell VEGF. Conditioned media from cells transduced with this rAAV or with rAAV-expressing endostatin or angiostatin inhibited capillary endothelial cell proliferation in vitro. Antiangiogenic rAAVs may offer a novel gene therapy approach to undermining tumor neovascularization and cancer progression. PMID- 9865721 TI - Up-regulation of hypoxia-inducible factor-1alpha is not sufficient for hypoxic/anoxic p53 induction. AB - Oxygen-deprived regions of a solid tumor can induce tumor suppressor p53 expression and hence select for p53-mutant tumor cells with diminished apoptotic potential. It has been proposed that the hypoxia-inducible factor-1 (HIF-1) alpha subunit binds to p53 and protects it from proteasomal degradation. However, we found that hypoxic conditions that strongly induce HIF-1-dependent endogenous gene expression as well as HIF-1alpha protein neither induce p53-dependent gene expression nor p53 protein. The iron chelator deferoxamine induced both HIF 1alpha and p53, but p53 up-regulation could still be detected in HIF-1alpha deficient cells, suggesting that mechanisms other than HIF-1alpha activation contribute to oxygen-regulated p53 induction. PMID- 9865722 TI - maspin suppresses the invasive phenotype of human breast carcinoma. AB - The recently discovered tumor suppressor gene maspin has been shown to inhibit tumor cell motility, invasion, and metastasis in breast cancer by our laboratories. Nonetheless, the exploitation of maspin as a potential diagnostic and/or therapeutic tool has remained limited due to the lack of knowledge concerning its molecular and biological mechanism(s) of action. The work reported here demonstrates that recombinant maspin (rMaspin) has the ability to induce higher cell surface levels of alpha5- and alpha3-containing integrins and reduced levels of alpha2-, alpha4-, alpha6-, alpha(v)-, and some beta1-containing integrins in the metastatic human breast carcinoma cell line MDA-MB-435 concomitant with its ability to inhibit the invasive process in vitro. Furthermore, treatment of MDA-MB-435 cells with rMaspin results in the selective adhesion of the cell to a fibronectin matrix and conversion from a fibroblastic to a more epithelial-like phenotype. In addition, the ability of rMaspin to inhibit the invasive process can be abrogated with a blocking antibody to the alpha5beta1 integrin, which diminishes the ability of the cells to invade through a fibronectin matrix-containing barrier in vitro. Taken together, these data address the hypothesis that rMaspin reduces the invasive phenotype of MDA-MB-435 cells by altering their integrin profile, particularly alpha5, which in turn converts these cells to a more benign epithelial phenotype, with less invasive ability. These data provide new insights into the biological significance of this tumor suppressor gene found in normal mammary epithelium and may form the basis of novel therapeutic strategies in the management of breast carcinoma. PMID- 9865723 TI - Potentiation of the antitumor effect of ionizing radiation by brief concomitant exposures to angiostatin. AB - Angiostatin, a proteolytic fragment of plasminogen, inhibits the growth of primary and metastatic tumors by suppressing angiogenesis. When used in combination with ionizing radiation (IR), angiostatin demonstrates potent antitumor synergism, largely caused by inhibition of the tumor microvasculature. We report here the temporal interaction of angiostatin and IR in Lewis lung carcinoma (LLC) tumors growing in the hind limbs of syngeneic mice. Tumors with an initial mean volume of 510 +/- 151 mm3 were treated with IR alone (20 Gy x 2 doses on days 0 and 1), angiostatin alone (25 mg/kg/day divided twice daily) on days 0 through 13, or a combination of the two as follows: (a) IR plus angiostatin (days 0 through 13); (b) IR plus angiostatin (days 0 and 1); and (c) IR followed by angiostatin beginning on the day after IR completion and given daily thereafter (days 2 through 13). By day 14, tumors in untreated control mice had grown to 6110 +/- 582 mm3, whereas in mice treated with: (a) IR alone, tumors had grown to 2854 +/- 338 mm3 (P < 0.05 compared with untreated controls); and (b) angiostatin alone, tumors had grown to 3666 +/- 453 mm3 (P < 0.05 compared with untreated controls). In combined-treatment groups, in mice treated with: (a) IR plus longer-course angiostatin, tumors reached 2022 +/- 282 mm3 (P = 0.036 compared with IR alone); (b) IR followed by angiostatin, tumors reached 2677 +/- 469 mm3 (P > 0.05 compared with IR alone); and (c) IR plus short-course angiostatin, tumors reached 1032 +/- 78 mm3 (P < 0.001 compared with IR alone). These findings demonstrate that the efficacy of experimental radiation therapy is potentiated by brief concomitant exposure of the tumor vasculature to angiostatin. PMID- 9865724 TI - Serial analysis of gene expression in non-small cell lung cancer. AB - We used the serial analysis of gene expression (SAGE) method to systematically analyze transcripts present in non-small cell lung cancer. Over 226,000 SAGE tags were sequence analyzed from two independent primary lung cancers and two normal human bronchial/tracheal epithelial cell cultures. A total of 226,000 SAGE tags were sequence identified, representing 43,254 unique transcripts. Comparison of the tags present in the tumor with those identified in the normal tissue revealed 175 transcript tags that were overrepresented in the normal tissue and 142 tags that were overexpressed in the tumor by 10-fold or more. Northern hybridization was performed on 15 of the most abundantly expressed tags identified in the tumors. These tags were derived from either a known gene or a matched expressed sequence tag clone. The transcripts for 3 of the 15 genes, PGP 9.5, B-myb, and human mutT, were abundantly expressed in primary lung cancers (10 of 18, 15 of 18, and 6 of 12 tumors, respectively). In contrast, the presence of PGP9.5 and B myb was much less frequent in primary tumors derived from other tissue origins. These results suggest that at least a portion of the transcripts identified by SAGE are frequently associated with lung cancer, and that their overexpression may contribute to lung tumorigenesis. The identification and further characterization of genes generated by SAGE should provide potential new targets for the diagnosis, prognosis, and therapy of lung cancer. PMID- 9865725 TI - Modulation of aromatase expression in the breast tissue by ERR alpha-1 orphan receptor. AB - We have previously identified a silencer element (S1) that is situated between promoters I.3 and II of the human aromatase gene and that down-regulates the action of these promoters. We recently applied the yeast one-hybrid approach to screen a human breast tissue hybrid cDNA expression library for genes encoding the proteins binding to the silencer region. Most proteins identified from this approach belong to the nuclear receptor superfamily. Fifty % of the positive clones encode for ERR alpha-1, and other positive clones include EAR-2, EAR-3 (COUP-TF1), RAR gamma, and p120E4F. Because ERR alpha-1 was found to be the major protein interacting with S1, we decided to examine the regulatory action of ERR alpha-1 on promoter I.3 of the human aromatase gene. Using a reporter plasmid that includes the aromatase genomic fragment containing promoter I.3 and S1, ERR alpha-1 was found to have a positive regulatory function in breast cancer SK-BR-3 cells. Gel mobility shift assays have confirmed that ERR alpha-1 binds to S1 in a dose-dependent manner, and DNase I footprinting analysis has revealed that ERR alpha-1 binds to a region, 5'-AAGGTCAGAAAT-3', which is within S1 and between 96 and 107 bp relative to the transcriptional start site of promoter I.3. In addition, despite the fact that the nuclear receptor SF1 was shown previously to bind to the same site and to mediate a cAMP response in ovary, our yeast one hybrid screening did not find any SF-1 clones. Gel mobility shift assays further revealed that SF-1 can bind to the silencer element with an affinity comparable with ERR alpha-1. Because our reverse transcription-PCR analysis was not able to detect SF1 mRNA in breast cancer tissue or in SK-BR-3 cells, it is thought that SF1 protein is not expressed in breast cancer tissue. Two ERR alpha-1 RNA variants with differences at the 5'-end have been reported. Our reverse transcription-PCR analysis identified the shorter variant in 28 of 32 breast tumor specimens and the longer variant in only 1 specimen. In addition, the shorter variant was detected in breast cancer SK-BR-3 cells as well as in a breast tumor fibroblast line WS3TF. The results suggest that ERR alpha-1 is one of the nuclear proteins interacting with S1 in breast cancer tissue. It is thought that the silencer element in the human aromatase gene may function differently in different tissues because of distinct expression patterns of transcription factors. PMID- 9865727 TI - Resveratrol inhibits transcription of CYP1A1 in vitro by preventing activation of the aryl hydrocarbon receptor. AB - Resveratrol, a compound present in a variety of plants, was recently shown to have potent chemopreventive activity against aryl hydrocarbon-induced tumorigenesis in mice. Therefore, in the present study, we examined the effect of resveratrol on the function of the aryl hydrocarbon receptor (AHR) and the transcription of CYP1A1 in human HepG2 hepatoma cells. Resveratrol inhibited the increase in cytochrome P450 (CYP) 1A1 mRNA caused by the AHR ligand 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) in a concentration-dependent manner. The induction of transcription of an aryl hydrocarbon-responsive reporter vector containing the CYP1A1 promoter by TCDD was likewise inhibited by resveratrol. Resveratrol also inhibited the constitutive level of CYP1A1 mRNA and reporter vector transcription in HepG2 cells. The increase in CYP1A1 enzyme activity induced by TCDD was inhibited by resveratrol. Resveratrol prevented the TCDD induced transformation of the cytosolic AHR to its nuclear DNA-binding form. However, resveratrol had no effect on the binding of TCDD to the cytosolic AHR. These data demonstrate that resveratrol inhibits CYP1A1 expression in vitro, and that it does this by preventing the binding of the AHR to promoter sequences that regulate CYP1A1 transcription. This activity may be an important part of the chemopreventive activity of resveratrol. PMID- 9865726 TI - High-throughput assay for G2 checkpoint inhibitors and identification of the structurally novel compound isogranulatimide. AB - Treatment of cancer cells lacking p53 function with G2 checkpoint inhibitors sensitizes them to the toxic effects of DNA damage and has been proposed as a strategy for cancer therapy. However, few inhibitors are known, and they have been found serendipitously. We report the development of a G2 checkpoint inhibition assay that is suitable for high-throughput screening and its application to a screen of 1300 natural extracts. We present the isolation of a new G2 checkpoint inhibitor, the structurally novel compound isogranulatimide. In combination with gamma-irradiation, isogranulatimide selectively kills MCF-7 cells lacking p53 function. PMID- 9865728 TI - Dietary modulation of carcinoma development in a mouse model for human familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) is caused by a dominant mutation in the adenomatous polyposis coli (APC) gene. Individuals with FAP progressively develop adenomas and carcinomas of the colon and rectum. We developed a mouse model for this disorder by genetically modifying the Apc gene. The resulting mice Apc1638 progressively develop neoplasms in the colon and remainder of the gastrointestinal tract. In this study when Apc1638 mice were fed a Western-style diet, they developed an increased incidence of the end point of carcinomas and number of invasive tumors. The findings therefore demonstrated dietary modulation of carcinoma incidence in mice with a targeted mutation providing a model for the study of gene-environment interactions in cancer. PMID- 9865729 TI - Androgen receptor expression in androgen-independent prostate cancer is associated with increased expression of androgen-regulated genes. AB - The human prostate cancer (CaP) xenograft, CWR22, mimics human CaP. CWR22 grows in testosterone-stimulated nude mice, regresses after castration, and recurs after 5-6 months in the absence of testicular androgen. Like human CaP that recurs during androgen deprivation therapy, the recurrent CWR22 expresses high levels of androgen receptor (AR). Immunohistochemical, Western, and Northern blot analyses demonstrated that AR expression in the androgen-independent CWR22 is similar to AR expression in the androgen-dependent CWR22 prior to castration. Expression of prostate-specific antigen and human kallikrein-2 mRNAs, two well characterized androgen-regulated genes in human CaP, was androgen dependent in CWR22. Despite the absence of testicular androgen, prostate-specific antigen and human kallikrein-2 mRNA levels in recurrent CWR22 were higher than the levels in regressing CWR22 tumors from 12-day castrate mice and similar to those in the androgen-stimulated CWR22. Other AR-regulated genes followed a similar pattern of expression. Differential expression screening identified androgen regulation of alpha-enolase and alpha-tubulin as well as other unknown mRNAs. Insulin-like growth factor binding protein-5, the homeobox gene Nkx 3.1, the AR coactivator ARA-70, and cell cycle genes Cdk1 and Cdk2 were androgen regulated in CWR22. In recurrent CWR22, the steady-state levels of all these AR-dependent mRNAs were similar to those in the androgen-stimulated CWR22, despite the absence of testicular androgen. Expression of AR and AR-regulated genes in the androgen deprived recurrent CWR22 at levels similar to the androgen-stimulated CWR22 suggests that AR is transcriptionally active in recurrent CWR22. Induction of these AR-regulated genes may enhance cellular proliferation in relative androgen absence but through an AR-dependent mechanism. Alternatively, in androgen independent tumors, induced expression of the AR-regulated gene network might result from a non-AR transcription control mechanism common to these genes. PMID- 9865730 TI - Tumor necrosis factor alpha-mediated tumor regression by the in vivo transfer of genes into the artery that leads to tumors. AB - We report that tumor necrosis factor (TNF) alpha induced a strong antitumor immune reaction when it was produced in arteries leading to tumors by gene transfer in vivo. We used a mouse model carrying a sarcoma-180 tumor in the right footpad and injected the fusogenic liposomes encapsulating the human TNF-alpha gene into the right femoral artery. Under this condition, human TNF-alpha was detected only in the artery leading to the tumor and in the tumor. There was a significant regression in tumor growth when the TNF-alpha gene was delivered into the right femoral artery, with 4 of 11 mice completely cured. No regression was observed when the TNF-alpha gene was delivered into the left femoral artery or into the tumor or when the luciferase gene was administered. Tumor regression was inhibited by the injection of anti-TNF-alpha, anti-CD4, or anti-CD8 monoclonal antibody, and CD8+ T cells accumulated in the tumors of TNF-alpha-treated mice. These results suggest that TNF-alpha expressed locally in the arteries leading to tumors efficiently suppresses tumor growth through reinforcement of an antitumor immune reaction. The significance of this phenomenon for cancer gene therapy was discussed. PMID- 9865731 TI - Enhanced tumor cell killing in the presence of ganciclovir by herpes simplex virus type 1 vector-directed coexpression of human tumor necrosis factor-alpha and herpes simplex virus thymidine kinase. AB - Past studies have documented the promise of herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) suicide gene therapy as a potential antitumor treatment. HSV-TK converts the pro-drug ganciclovir (GCV) into a toxic nucleotide analogue, the incorporation of which into cellular DNA blocks cell proliferation. In this report, we have examined the hypothesis that the effectiveness of HSV-TK suicide gene therapy can be enhanced by coexpression of the antitumor cytokine human tumor necrosis factor-alpha (TNF-alpha) from the same replication-defective HSV-1 vector. In vitro testing demonstrated that TNF-alpha expression from this vector potentiated the killing of both TNF-alpha-sensitive L929 tumor cells and TNF alpha-resistant U-87 MG cells in the presence of GCV. Furthermore, treatment of established intradermal L929 tumors in vivo with the TNF-alpha/TK vector and GCV resulted in prolonged animal survival compared with treatment with parental HSV TK vector in the presence or absence of GCV. Treatment of intracerebral U-87 MG tumors showed a clear benefit of TK therapy, but a significant further increase in survival using the TNF-alpha vector could not be demonstrated. We found that potentiation of cell killing in vitro required intracellular TNF-alpha because purified protein added to the culture medium of cells infected with HSV-TK vector failed to have the same effect. Accordingly, potentiation in vivo should depend on efficient infection, but immunohistochemical analysis indicated that virus administration by U-87 MG intratumoral injection was inadequate, resulting in an estimated <1% infection of all tumor cells. Moreover, the majority of infected tumor cells were localized at the tumor margin. Together, these results suggest that TNF-enhanced tk gene therapy should provide a useful treatment for TNF-alpha sensitive tumors and perhaps also for TNT-alpha-resistant tumors if vector delivery can be improved to increase the percentage of transduced tumor cells. PMID- 9865732 TI - Differential susceptibility of primary and established human glioma cells to adenovirus infection: targeting via the epidermal growth factor receptor achieves fiber receptor-independent gene transfer. AB - Adenovirus (Ad) vectors are promising for gene therapy of glioma due to their ability to achieve efficient gene transfer upon intratumoral administration. Yet in this context, Ad mediates widespread gene transfer to both tumor and surrounding parenchyma. Ad entry is dependent upon the expression of fiber receptors, such as coxsackie/adenovirus receptor, and alpha(v) integrins on the target cells for binding and internalization, respectively. We hypothesized that the susceptibility of human gliomas to Ad would likely be heterogeneous due to variable expression of these receptors. It was found that established human glioma cell lines exhibited differential susceptibility to Ad-mediated gene transfer, which correlated directly with the level of radiolabeled Ad binding and with the expression of coxsackie/adenovirus receptor but not with the expression of alpha(v) integrins. To circumvent the lack of fiber receptors and to target Ad gene transfer specifically to tumor cells, we used a bispecific antibody conjugate to ablate Ad binding to fiber receptors and retarget binding to the epidermal growth factor receptor (EGFR), a tumor-associated marker negligibly expressed in normal, mitotically quiescent neural tissues. The results demonstrate that EGFR-targeted Ad gene transfer was EGFR specific and independent of fiber-fiber receptor interactions. Furthermore, EGFR targeting significantly enhanced Ad gene delivery to 7 of 12 established glioma cell lines and to 6 of 8 cultured primary gliomas. Interestingly, EGFR-targeted Ad gene transfer did not correlate with EGFR expression across cell lines, suggesting the importance of other factors. This study establishes that fiber receptor expression limits the utility of Ad vectors for gene transfer to glioma cells and suggests that targeting Ad via EGFR may prove valuable for tumor-specific gene transfer to high grade gliomas. These findings have key relevance in the context of Ad vector based approaches for glioma gene therapy. PMID- 9865733 TI - Modeling of the time-dependency of in vitro drug cytotoxicity and resistance. AB - For potential clinical extrapolation of in vitro findings, it is of interest to relate the measured effect of an anticancer agent to concentration and exposure time. The Hill model (A. V. Hill, J. Physiol., 40: iv-vii, 1910) is commonly used to describe pharmacodynamic (PD) effects, including drug-induced growth inhibition of cancer cells in vitro. The IC(X)n x T = k relationship, in which IC(X) is the concentration of agent required to reduce cell growth by X%, T is the exposure time, and n and k are estimable parameters, was first applied to bacterial disinfectant action and then was successfully used to model anticancer drug potency as a function of exposure time (D. J. Adams, Cancer Res., 49: 6615 6620, 1989). Our goal was to create a new global PD modeling paradigm to facilitate the quantitative assessment of the growth-inhibitory effect of anticancer agents as a function of concentration and exposure time. Wild-type human ovarian A2780 and ileocecal HCT-8 carcinoma cells and sublines that were resistant to cisplatin (A2780/CP3), doxorubicin (A2780/DX5B), and raltitrexed (RTX) (HCT-8/DW2) were exposed to various anticancer agents, cisplatin, doxorubicin, paclitaxel, trimetrexate, RTX, methotrexate, and AG2034, for periods ranging from 1 to 96 h. Cell growth inhibition was measured with the sulforhodamine B protein dye assay. Patterns of time-dependency of drug potency, slope of the concentration-effect curves, and relative degree of resistance were characterized. Empirical mathematical expressions were built into a global concentration-time-effect model. The global PD model was then fit to the concentration-time-effect data with iteratively reweighted nonlinear regression. Under specific treatment conditions, the examination of the slope and the shape of the concentration-effect curves revealed a large heterogeneity in drug response, e.g., shallow concentration-effect curve or double or triple Hill "roller coaster" concentration-effect curve. These patterns, which were observed at intermediate exposure times in parental and resistant cells for paclitaxel and trimetrexate or only in resistant HCT-8/DW2 cells for RTX, methotrexate, and AG2034, revealed mechanistic insights for the former cases but possible methodological artifacts for the latter cases. The comprehensive PD modeling of the cytotoxic effect of anticancer agents showed that it was possible to modulate drug effect, response heterogeneity, and drug resistance by altering the time of exposure to the agents. This approach will be useful for: (a) describing complex concentration-time-effect surfaces; (b) refining biological interpretations of data; (c) providing insights on mechanisms of drug action and resistance; and (d) generating leads for clinical use of anticancer drugs. PMID- 9865734 TI - Transcriptional suppression of multidrug resistance-associated protein (MRP) gene expression by wild-type p53. AB - Multidrug resistance is a major obstacle to the success of cancer chemotherapy. The multidrug resistance-associated protein (MRP) has been shown to confer multidrug resistance. To study MRP gene expression at the transcriptional level, we have fused the MRP gene promoter with the luciferase reporter gene and studied its regulation. Cotransfection of MRP promoter constructs with p53 expression plasmids in p53-null human H1299 and mouse (10)1 cells demonstrated that the wild type (wt) p53 markedly suppressed MRP promoter activity, whereas mutant p53 had little inhibitory effect. Transfections using 5' deletion mutant constructs of the MRP promoter showed that inhibition of the promoter activity by wt p53 mainly resided in the region from -91 to +103 bp, where several Sp1 transcription factor binding sites are localized. Cotransfection of the MRP promoter into Drosophila SL2 cells with an Sp1 expression vector increased the promoter activity in a dose related manner up to approximately 200-fold. The stimulation of MRP promoter activity by Sp1 was attenuated by the cotransfection of a wt p53-expression plasmid. Furthermore, we have determined that endogenous MRP mRNA levels were down-regulated by restoration of wt p53-expression in a human lung cancer cell line. The relevance of MRP regulation in drug resistance was studied in a drug resistant cell line, CEM/VM-1-5, that is approximately 140-fold more resistant to the epipodophyllotoxin, teniposide (VM-26), than the parental CEM cells. CEM/VM-1 5 cells express a much higher amount of MRP mRNA and protein than CEM cells, indicating that the resistant phenotype is at least partly due to increased MRP production. Transient transfection of the promoter constructs revealed that CEM/VM-1-5 cells had higher (7-fold) MRP promoter activity than CEM cells. Cotransfection of a wt p53-expression plasmid caused a reduction of MRP promoter activity in both CEM and CEM/VM-1-5 cells, but the inhibition was more than double in CEM/VM-1-5 cells compared with CEM cells. Our results demonstrated that wt p53 acts as a negative regulator of MRP gene transcription, at least in part by diminishing the effect of a powerful transcription activator Sp1. Therefore, a loss of wt p53 function and/or an increase in Sp1 activity in tumor cells could contribute to an up-regulation of the MRP gene. PMID- 9865735 TI - Constitutive expression of cellular retinoic acid binding protein II and lack of correlation with sensitivity to all-trans retinoic acid in acute promyelocytic leukemia cells. AB - The up-regulation of cellular retinoic acid binding protein-II (CRABP-II) has been invoked as an important mechanism of clinically acquired resistance to all trans retinoic acid (RA) therapy in acute promyelocytic leukemia (APL). To test this hypothesis, we used quantitative reverse transcription-PCR and fast performance liquid chromatography procedures to examine the levels of CRABP-II mRNA and RA binding activity in APL patient samples. We found that CRABP-II mRNA in APL cells from pretreatment patients (n = 36) was constitutively expressed at relatively high levels (median, 0.92; range, 0.16-4.13) relative to the level in CRABP-H protein-expressing NB4 cells (arbitrarily set at 1.0 unit). Consistent with this finding, the RA binding activity of CRABP in APL cells from three pretreatment cases (range, 27.2-53.2 fmol/mg protein) was similar to that of NB4 cells (22.6 +/- 5.4 fmol/mg protein). Furthermore, in the pretreatment samples, there was no association between CRABP-H mRNA expression level and APL cellular sensitivity to RA-induced differentiation in vitro. After 45 days of remission induction therapy on Eastern Cooperative Oncology Group protocol E2491, CRABP-II mRNA was modestly increased from day 0 values in patients treated with either RA (median increase, 0.41) or chemotherapy (median increase, 0.56), and there was no significant difference between the two treatment groups (P = 0.91). In patients studied after relapse from RA therapy (n = 7), there was a significant decline in APL cell sensitivity to RA-induced differentiation in vitro compared with patients after relapse from chemotherapy (n = 5; P = 0.015-0.055 at three RA concentrations tested), but in the RA relapse cases, there was no change from pretreatment levels of CRABP-II mRNA (median, 0.98) or, in three relapse cases studied, of RA protein binding activity (range, 22.1-70.7 fmol/mg protein). Taken together, our data strongly imply that variations in CRABP-II expression and RA binding activity are not causally related to the development of clinically acquired APL cellular RA resistance, but rather, they suggest that constitutive expression of CRABP-II could have a facilitative role in the response of APL cells to RA. PMID- 9865736 TI - Light-induced photoactivation of hypericin affects the energy metabolism of human glioma cells by inhibiting hexokinase bound to mitochondria. AB - Glucose-dependent energy required for glioma metabolism depends on hexokinase, which is mainly bound to mitochondria. A decrease in intracellular pH leads to a release of hexokinase-binding, which in turn decreases glucose phosphorylation, ATP content, and cell proliferation. Thus, intracellular pH might be a target for therapy of gliomas, and a search for agents able to modulate intracellular pH was initiated. Hypericin, a natural photosensitizer, displays numerous biological activities when exposed to light. Its mechanism and site of action at the cellular level remain unclear, but it probably acts by a type II oxygen-dependent photosensitization mechanism producing singlet oxygen. Hypericin is also able to induce a photogenerated intracellular pH drop, which could constitute an alternative mechanism of hypericin action. In human glioma cells treated for 1 h with 2.5 microg/ml hypericin, light exposure induced a fall in intracellular pH. In these conditions, mitochondria-bound hexokinase was inhibited in a light- and dose-dependent manner, associated with a decreased ATP content, a decrease of mitochondrial transmembrane potential, and a depletion of intracellular glutathione. Hexokinase protein was effectively released from mitochondria, as measured by an ELISA using a specific anti-hexokinase antibody. In addition to decreased glutathione, a response to oxidative stress was confirmed by the concomitant increase in mRNA expression of gamma-glutamyl cysteine synthetase, which catalyzes the rate-limiting step in overall glutathione biosynthesis, and is subject to feedback regulation by glutathione. Hypericin also induced a dose- and light-dependent inhibition of [3H]thymidine uptake and induced apoptosis, as demonstrated by annexin V-FITC binding and cell morphology. This study confirmed the mitochondria as a primary target of photodynamic action. The multifaceted action of hypericin involves the alteration of mitochondria-bound hexokinase, initiating a cascade of events that converge to alter the energy metabolism of glioma cells and their survival. In view of the complex mechanism of action of hypericin, further exploration is warranted in a perspective of its clinical application as a potential phototoxic agent in the treatment of glioma tumors. PMID- 9865737 TI - Host-mediated antibody-dependent cellular cytotoxicity contributes to the in vivo therapeutic efficacy of an anti-CD7-saporin immunotoxin in a severe combined immunodeficient mouse model of human T-cell acute lymphoblastic leukemia. AB - We have investigated the anti-leukemia effect that is exerted by the murine anti CD7 antibody HB2 in a severe combined immunodeficient (SCID) mouse model of human T-cell acute lymphoblastic leukemia (T-ALL) and determined the contribution that this antibody effect makes to the therapeutic potency of a saporin immunotoxin (IT) constructed with the same antibody. The anti-leukemia effect is not exerted through complement-mediated lysis or through direct growth-inhibitory signaling after binding of antibody to the CD7 molecule on the T-ALL cell surface but rather through antibody-dependent cellular cytotoxicity (ADCC). Thus, the in vivo depletion of SCID mice of their natural killer cells almost completely abolishes the therapeutic effect of native HB2 anti-CD7 antibody and moreover significantly reduces the in vivo therapeutic performance of the anti-CD7 HB2-SAPORIN IT. Furthermore, an IT constructed with the F(ab')2 fragment of the same anti-CD7 antibody (HB2-F(ab')2-SAPORIN), which is incapable of recruiting natural killer cells, performed significantly less well therapeutically than HB2-SAPORIN IT. There was also a significant improvement in the therapeutic performance of the HB2-F(ab')2-SAPORIN IT in SCID-HSB-2 mice when used in combination with intact HB2 antibody, presumably through restoration of an ADCC attack on the target HSB 2 cell. These combined data indicate that ADCC in the SCID mouse does contribute additively together with toxin to the in vivo therapeutic potency of the HB2 SAPORIN IT directed against this human T-ALL cell line and that this has potentially important implications for the utility of this and other related classes of immunotherapeutic in human therapy. PMID- 9865738 TI - The induction of in vivo proliferation of long-lived CD44hi CD8+ T cells after the injection of tumor cells expressing IFN-alpha1 into syngeneic mice. AB - The tumorigenicity of transplantable tumor cells in mice is reduced by transduction with cytokine genes, including IFN-alpha and interleukin (IL) 12. Although T cells are considered important in tumor rejection, the mechanism by which genetically modified tumor cells stimulate the immune system has not been examined. In this study, the in vivo proliferation of T-cell subsets in mice transplanted with cytokine-producing syngeneic tumor cells was assessed by administering the DNA precursor bromodeoxyuridine. The injection of viable cells producing IFN-alpha or IL-12 caused a marked proliferation of CD8+ T lymphocytes in both the spleen and lymph nodes. Proliferation was most prominent among memory phenotype CD44hi CD8+ T cells. In contrast, proliferation of CD8+ T cells did not occur in mice injected with control cells or with cells expressing IL-4, granulocyte colony-stimulating factor, or IFN-gamma. Pulse-chase studies in mice injected with IFN-alpha-producing cells showed that a proportion of proliferating CD8+ T cells survived for at least 70 days, suggesting that long-lived memory cells are induced using such an approach. In summary, these results, together with previous studies on the host immune reactivity triggered by the injection of tumor cells expressing IFN-alpha, represent a strong rationale for considering IFN-alpha as a powerful T-cell adjuvant for the generation of more effective cancer vaccines. PMID- 9865739 TI - Direct identification of major histocompatibility complex class I-bound tumor associated peptide antigens of a renal carcinoma cell line by a novel mass spectrometric method. AB - Melanoma and renal cell carcinoma (RCC) are thought to be the most immunogenic human tumors. Presently a series of tumor-specific peptides of melanoma is being tested in clinical trials with different immunotherapy protocols. In contrast, only one decameric peptide (SPSSNRIRNT) derived from one (ORF2) of three possible open reading frames (ORFs) of a gene named RAGE (Renal tumor AntiGEn) was shown to be the target for tumor-specific CTLs on renal carcinoma cells. One reason for the lack of identification of tumor antigens on RCC compared with melanoma may be the difficulty in generating tumor-specific CTLs as screening instruments. Therefore, our approach was directly to isolate and identify peptides bound to HLA class I molecules of the HLA-A2 and -B8 homozygous RCC line A-498. High performance liquid chromatography-fractionated peptides eluted with acid from immunoaffinity-purified HLA class I-peptide complexes were sequenced and identified for the first time by the novel and highly sensitive mass spectrometric method matrix-assisted laser desorption ionization-post source decay (MALDI-PSD) from minute amounts of 100 fmol to 1.5 pmol of the fractionated peptide samples. Fourteen peptide sequences first deduced from interpretations of the mass spectra were also shown to fulfill other reliability criteria such as matching the mass spectra of the respective synthetic peptides. Some peptides were identified to be derived from genes preferentially activated in malignant tissues or resulted from a possibly mutated gene. The most promising candidate for a CTL epitope is a decameric peptide (PASKKTDPQK) derived from another possible ORF (ORF5) of the RAGE gene and probably presented in association with HLA-B8. This peptide was synthesized and used for the in vitro induction of CTLs that lysed the A-498 cells and another HLA-B8-positive RCC line significantly more strongly than either other RAGE-positive but HLA-B8-negative RCC lines or K562 cells. Sensitive sequencing by MALDI-PSD thus may provide a powerful method of identifying potentially tumor-specific and HLA-restricted antigens, even on native malignant cells and tissues. PMID- 9865740 TI - IgG2a induced by interleukin (IL) 12-producing tumor cell vaccines but not IgG1 induced by IL-4 vaccine is associated with the eradication of experimental metastases. AB - We evaluated whether antibody response correlates with tumor therapy by cytokine gene-modified tumor cell vaccines. To characterize the antibody (Ab) response against a known antigen, colon carcinoma C26 cells and C26 variants engineered to produce interleukin (IL) 12 or IL-4 were further transduced to express the human tumor-associated antigen gp38 folate receptor (FR) alpha. Irradiated IL-12- and IL-4-producing C26/FR alpha cell vaccines cured 50 and 30% of mice bearing C26/FR alpha lung micrometastases. Treatment induced a rapid, CD4-dependent Ab production dominated by IgG2a and IgG1 in response to the IL-12 or IL-4 vaccine, respectively. In contrast, untreated tumor-bearing mice showed a late serological response dominated by IgM. Anti-FR alpha IgG1 and IgG2a were able to suppress tumor metastases upon passive transfer in vivo. Sera from mice cured by the IL-12 vaccine displayed a higher binding activity, a higher anti-FR alpha IgG2a content, and a higher complement-mediated tumor cell lysis in vitro compared to the sera from nonresponder mice. Such a correlation was not found in the sera of mice treated with the IL-4 vaccine. These data indicate that cytokine-producing tumor cell vaccines strongly influence antibody response, and that in the case of the IL-12-based vaccine, the Ab titer correlates with the therapeutic response, thus suggesting its use for monitoring the outcome of vaccination in cancer patients. PMID- 9865741 TI - Altered expression of heterogenous nuclear ribonucleoproteins and SR factors in human colon adenocarcinomas. AB - Alternative splicing is part of the expression program of a wide number of genes implicated in cell growth and differentiation. Although the occurrence of inappropriate alternative splicing in tumors has started to emerge, the underlying molecular mechanisms have been, thus far, largely unexplored. We have investigated the alternative splicing pattern of the CD44 gene in specimens of nonfamilial colon adenocarcinomas at different stages of tumor progression. In the same patients, we have assessed by Northern blotting analysis the mRNA levels of different heterogeneous nuclear ribonucleoproteins and SR factors, all involved in pre-mRNA splicing and, more in general, in mRNA maturation. The results of this analysis highlight a general rule for the mode of splicing of the CD44 pre-mRNA. Moreover, we found that the mRNA levels of different SR proteins in tumor specimens are different from, and usually lower than, those detected in samples of nonpathological tissue adjacent to the tumor. Quantitative analysis demonstrates that, in tumors, the mRNA levels of ASF, SRp40, SRp55, and SRp75, when normalized to those of heterogeneous nuclear ribonucleoprotein A1, are lower than those of SRp20 and SRp30. Interestingly, this reduction is more drastic in patients showing a more altered CD44 splicing pattern and seems to be related to the propensity to develop metastases. PMID- 9865742 TI - Liver regeneration and alpha-fetoprotein messenger RNA expression in the retrorsine model for hepatocyte transplantation. AB - Recently, we described a new model for hepatocyte transplantation with nearly total replacement of the liver by exogenous hepatocytes (E. Laconi et al., Am. J. Pathol., 153: 319-329, 1998). The model is based on the mitoinhibitory effect of the pyrrolizidine alkaloid retrorsine on hepatocytes in the resident liver while transplanted hepatocytes proliferate. In this study, we exploit this novel approach to address the important and controversial issue of whether hepatocytes, when proliferating extensively, undergo dedifferentiation and give rise to foci of undifferentiated hepatocytes. Genetically marked hepatocytes (isolated from normal Dipeptidyl peptidase IV+ Fischer 344 rats) were delivered intraportally (2 x 10(6) cells) into the liver of retrorsine-treated Dipeptidyl peptidase IV- mutant Fischer 344 rats in conjunction with partial hepatectomy. Transplanted hepatocytes were detected histochemically or immunohistochemically, and cell proliferation was studied by in situ hybridization for histone-3 mRNA. Expression of alpha-fetoprotein (AFP) mRNA, a marker of hepatocyte dedifferentiation, was also revealed by in situ hybridization. One day after partial hepatectomy and hepatocyte transplantation, endogenous hepatocytes and oval cells expanding in the liver expressed histone-3 mRNA (cells had entered S phase); 2 days later, transplanted hepatocytes and nonparenchymal cells also expressed histone-3 mRNA. Although the majority of endogenous hepatocytes did not divide and became arrested as quiescent megalocytes, the exogenous hepatocytes, as well as newly formed small hepatocytes, most probably derived from liver progenitor cells, underwent extensive proliferation. After 7-14 days, the nonparenchymal cells stopped proliferating, but transplanted hepatocytes and small endogenous hepatocytes continued to proliferate for 1 month, forming foci of dividing parenchymal cells. Although many of the hepatocytes in clusters were in S phase (histone-3 mRNA positive), none expressed AFP mRNA. In contrast, high expression of AFP mRNA was observed in proliferating oval and transitional cells, forming duct-like structures of cytokeratin-19-positive cells. From these studies, we conclude that hepatocyte proliferation in the adult liver is not associated with dedifferentiation. PMID- 9865743 TI - Clonal and chronological genetic analysis of multifocal cancers of the bladder and upper urinary tract. AB - Recent molecular genetic studies have suggested that multifocal urothelial cancers are derived from an identical progenitor cell. However, the clonal origin of multifocal urothelial cancers of a low-grade superficial type has not been fully defined. Using microsatellite markers, we examined genetic alterations at 20 loci on eight chromosomal arms (2q, 4p, 4q, 8p, 9p, 9q, 11p, and 17p) in 87 metachronous and/or synchronous multifocal urothelial cancers, which included 84 low-grade superficial papillary tumors from 29 patients. Judging from the patterns of loss of heterozygosity, microsatellite shifts, and the subchromosomal partial deletion, multifocal tumors in at least 20 (80%) of the 25 evaluable patients were considered to be derived from a single progenitor cell, although the possibility remained that multifocal tumors in a small subset of patients might develop from distinct progenitor cells due to field cancerization. In 13 of the 20 patients, a chronological genetic analysis was available: genetic heterogeneity was detected in 3 (23%) patients, and an apparent accumulated pattern of genetic alterations was detected in only 1 (8%) patient. In the 20 patients with multifocal tumors of an identical clonal origin, discordant microsatellite alterations were observed, with significantly lower frequencies on chromosome 9 compared to those on the other chromosomes tested. The results indicate that most multifocal low-grade superficial urothelial cancers are genetically stable despite their incidence of frequent recurrence, and genetic divergence occurs in a subset of patients. This heterotopic spread and genetic divergence may occur long before the clinical manifestation of multiplicity from a single transformed cell. These data support the previous view that heterotopic spread of transformed progenitor cells and genetic divergence occur after chromosome 9 alterations in most of low-grade superficial urothelial cancers. PMID- 9865744 TI - Simultaneous expression of Fas and nonfunctional Fas ligand in Ewing's sarcoma. AB - Fas-Fas ligand interactions play a central role in the regulation of the immune response. Fas ligand expression by tumors has been implicated in the abrogation of the host antitumor response by killing of Fas-positive effector lymphocytes. We have studied the presence and functional status of Fas and Fas ligand in Ewing's sarcoma. All Ewing's sarcoma cell lines tested expressed Fas on their surface. Three of the cell lines were readily killed after ligation of the Fas receptor. Four additional cell lines exhibited Fas-mediated apoptosis after preincubation with IFN-gamma and/or cycloheximide, whereas two cell lines were resistant to Fas-mediated killing. With regard to Fas ligand, all cell lines examined were positive for protein by immunoblot, and specificity was confirmed by reverse transcription-PCR. However, using flow cytometric analysis, Fas ligand could only be detected in Ewing's sarcoma cells after permeabilization. Furthermore, the cell lines were not capable of inducing apoptosis of Fas sensitive Jurkat cells. In addition, Ewing's sarcoma cell lines were able to serve as stimulators for the generation of cytotoxic effector lymphocytes and were susceptible to lysis by them. Therefore, Fas ligand is expressed in Ewing's sarcoma but is not functional, suggesting that Ewing's sarcoma is a potential target for immunotherapy. PMID- 9865745 TI - Reversal of a novel multidrug resistance mechanism in human colon carcinoma cells by fumitremorgin C. AB - We selected a human colon carcinoma cell line in increasing concentrations of mitoxantrone to obtain a resistant subline, S1-M1-3.2, with the following characteristics: profound resistance to mitoxantrone; significant cross resistance to doxorubicin, bisantrene, and topotecan; and very low levels of resistance to Taxol, vinblastine, colchicine, and camptothecin. This multidrug resistance (MDR) phenotype, which was not reversed by verapamil or another potent P-glycoprotein (Pgp) inhibitor, CL 329,753, was dependent, in part, upon an energy-dependent drug efflux mechanism. Pgp and the multidrug resistance protein (MRP) were not elevated in the resistant cells relative to the drug-sensitive parent, suggesting that resistance was mediated by a novel pathway of drug transport. A cell-based screen with S1-M1-3.2 cells was used to identify agents capable of circumventing this non-Pgp, non-MRP MDR. One of the active agents identified was a mycotoxin, fumitremorgin C. This molecule was extremely effective in reversing resistance to mitoxantrone, doxorubicin, and topotecan in multidrug-selected cell lines showing this novel phenotype. Reversal of resistance was associated with an increase in drug accumulation. The compound did not reverse drug resistance in cells with elevated expression of Pgp or MRP. We suggest that fumitremorgin C is a highly selective chemosensitizing agent for the resistance pathway we have identified and can be used as a specific pharmacological probe to distinguish between the diverse resistance mechanisms that occur in the MDR cell. PMID- 9865746 TI - Cytogenetic and fluorescence in situ hybridization characterization of chromosome 1 rearrangements in head and neck carcinomas delineate a target region for deletions within 1p11-1p13. AB - Cytogenetic analyses have revealed structural rearrangements of chromosome 1 in a large fraction of head and neck carcinomas (HNCA). These aberrations frequently affect chromosomal band 1p13 and the centromeric region, the latter often in the form of isochromosome i(1q) and whole-arm translocations. To delineate the critical region involved in rearrangements of proximal 1p, we have undertaken a more precise breakpoint mapping in 13 HNCAs, using metaphase fluorescence in situ hybridization with 11 yeast artificial chromosome (YAC) clones spanning 1p. All of the tumors had chromosome 1 changes at G-banding analyses. Fluorescence in situ hybridization showed that in almost all of the cases, at least one copy of chromosome 1 was affected by centromeric rearrangement. By the use of YAC clones mapped to juxtacentromeric regions and a centromere-specific alpha-satellite probe, we detected variable breakpoints in the whole-arm translocations. At the cytogenetic level, 1p13 rearrangements were frequent. However, molecular breakpoints within this band varied among the HNCAs tested. The lack of consistently rearranged chromosome segments indicates that the pathogenetically important consequence of 1p rearrangements in HNCAs is loss and/or gain of genes outside the breakpoint regions. In an assessment of the genomic imbalances, partial or complete overrepresentation of 1q was seen in eight cases. Loss of 1p material was also identified in eight cases; and in four of them, the deleted segments were too small to be discovered by G-banding analysis. The minimal overlapping deleted region was in the interval between YAC 959C4 (band p11-p12) and the centromere (p10). Our findings indicate that a target region potentially harboring tumor suppressor gene(s) crucial for HNCA is located within chromosomal bands 1p11-p13. PMID- 9865747 TI - Primary tumor size-dependent inhibition of angiogenesis at a secondary site: an intravital microscopic study in mice. AB - Some primary tumors are capable of suppressing the growth of their metastases by presumably generating antiangiogenic factors such as angiostatin. We hypothesized that the amount of inhibitor(s) released by a tumor increases with tumor growth. We tested this hypothesis by evaluating the relationship between the size of a primary tumor and its ability to inhibit angiogenesis at a secondary site. Furthermore, we characterized the effects of the primary tumor on physiological properties of newly formed vessels at the secondary site. Angiogenesis and physiological properties were measured using intravital microscopy of angiogenic vessels in the gels containing basic fibroblast growth factor placed into cranial windows of immunodeficient mice bearing human prostatic carcinoma (PC-3) in their flank. The PC-3 tumor inhibited angiogenesis in the gels, and surgical resection of tumor reversed this inhibition. The inhibition of angiogenesis 20 days after gel implantation (range, 0-83%) correlated positively (r = 0.625; P < 0.008) with the tumor size on the day of gel implantation (range, 19-980 mm3). The primary tumor also suppressed leukocyte-adhesion in angiogenic vessels, thus helping them evade the immune recognition. These results provide an additional rationale for combining antiangiogenic treatment with local therapies. PMID- 9865748 TI - Fas-mediated apoptosis in human prostatic carcinoma cell lines occurs via activation of caspase-8 and caspase-7. AB - We previously demonstrated that treatment with cycloheximide (CHX) converted the phenotype of Fas-resistant human prostatic carcinoma cell lines to Fas-sensitive and that resistance to Fas-mediated apoptosis was due to a dominant-negative protein(s). In this study, we investigated the sequential activation of caspase family members, to gain insight into the likely site of action of the suppressor protein(s). We did not find Tyr-Val-Ala-Aspase activity in any of the cell lines examined. Time-dependent Asp-Glu-Val-Aspase activity was detected during Fas mediated apoptosis in Fas-sensitive cell lines PC3 and ALVA31. Asp-Glu-Val-Aspase activity in Fas-resistant cell lines DU145 and JCA1, was detected only under combined treatment with CHX and anti-Fas agonistic mAb. In experiments with caspase inhibitors we show that Fas-mediated apoptosis in PC3 is mainly executed by the caspase-3 subfamily, but another member(s) of the caspase family may be involved in Fas-mediated apoptosis in ALVA31, DU145, and JCA1. Western blot analysis revealed that Fas-ligation activated caspase-7, but not caspase-3. The activated form of caspase-8 was detected in DU145 only after 4 h of simultaneous treatment with CHX and anti-Fas mAb, whereas in PC3 caspase-8 was found to be activated after 1 h of Fas-ligation. We have also found that treatment with staurosporin did not activate caspase-8, whereas staurosporin induced apoptosis at the same levels in both Fas-resistant and Fas-sensitive cell lines. These results suggest that an inhibitory protein(s), which suppresses apoptosis in Fas resistant cell lines, presumably acts at the apex of apoptotic cascade by preventing the activation of caspase-8. PMID- 9865750 TI - Interactive resources at the Archives Web site. PMID- 9865749 TI - Correspondence re: S. Fulda et al., Betulinic acid triggers CD95 (Apo1/Fas)- and p53-independent apoptosis via activation of caspases in neuroectodermal tumors. Cancer Res., 57: 4956-4964, 1997. PMID- 9865752 TI - An anatomical approach to glabellar rhytids. AB - OBJECTIVE: To identify surface landmarks that can serve as reference points to the underlying musculature in the treatment of glabellar rhytids. METHODS: Fifty cadaver hemibrows were dissected to assess the location, disposition, and relationships of the brow muscles, along with their variations at each of several consistent locations. Particular attention was paid to the corrugator supercilii, frontal belly of the frontalis, and procerus muscles. CONCLUSIONS: The information gained here may be applied to the pharmacological or surgical treatment of glabellar rhytids. Knowledge of the frequent location of the muscles involved, relative to easily identifiable surface landmarks, allows a more precise approach. PMID- 9865751 TI - Facial fracture classification according to skeletal support mechanisms. AB - OBJECTIVE: To construct, propose, and evaluate the usefulness of a new clinical facial fracture classification scheme to accurately denote, communicate, and compare facial fractures. DESIGN: A retrospective, consecutive sample study with application of the proposed classification scheme to denote maxillary and zygomatic fractures with computed tomography. SETTING: Metropolitan tertiary care trauma center. PATIENTS: A total of 213 consecutive adult patients with facial fractures evaluated by means of 2-dimensional computed tomography. RESULTS: The classification scheme is defined according to fractures of vertical buttresses and horizontal beams. The scheme uses 3 primary descriptors of laterality and support sites to denote the clinical pattern of the fractures. This scheme was accurately applied and sufficient to describe 87 midfacial fracture patterns in this study. In addition, 118 (98%) of 120 mock fracture patterns were correctly transcribed and reproducibly communicated among 12 participating physicians. CONCLUSIONS: This newly proposed facial fracture classification scheme provides a convenient, specific, descriptive, and reproducible method of denoting fracture patterns. This scheme may be used to accurately communicate and compare, in greater detail than permitted using current independent classification schemes, the essential site and degree-of-severity characteristics of facial fractures critical to their surgical reduction and reconstruction. The usefulness of this classification scheme in determining optimal methods and subsequent outcomes in midfacial fracture reduction requires further investigation. PMID- 9865753 TI - Prognostic significance of PCNA expression in laryngeal cancer. AB - OBJECTIVE: To assess the prognostic value of proliferating cell nuclear antigen (PCNA) in laryngeal carcinoma and its relation with other known prognostic clinicopathologic variables. DESIGN: A retrospective cohort study of 92 patients chosen randomly from patients treated between 1964 and 1993 with the diagnosis of laryngeal cancer. Prognostic factors including PCNA expression, grade, lymphovascular invasion, depth of tumor margins, neck metastasis, and clinical outcome were evaluated. SETTING: Hacettepe University Medical Faculty, Ankara, Turkey. PATIENTS: Eighty-five men and 7 women operated on for squamous cell carcinoma of the larynx were studied. Sixty-nine patients had total and 20 patients had partial laryngectomy with neck dissection, and 3 patients had endolaryngeal tumor excision. INTERVENTION: Hematoxylin and eosin-stained sections were reevaluated for grade, lymphovascular invasion, and depth of tumor margins; sections stained with monoclonal antibody against PC10 were examined for PCNA expression. RESULTS: The PCNA index correlated with grade, lymphovascular invasion, depth of tumor margins, neck metastasis, and local-regional recurrence. The PCNA index values of patients with occult metastasis were significantly higher than those of patients without metastasis (P=.006). CONCLUSIONS: The PCNA index is a more sensitive variable than grade in predicting tumor proliferation, occult lymph node metastasis, and prognosis. These results suggest that the PCNA index can be used in decision making for treatment and assessment of prognosis in laryngeal carcinomas. PMID- 9865754 TI - Basic fibroblast growth factor and insulinlike growth factor I support the growth of human septal chondrocytes in a serum-free environment. AB - OBJECTIVE: To determine if insulinlike growth factor I (IGF-I) and basic fibroblast growth factor (bFGF), individually or in combination, support the growth and viability of human septal chondrocytes in a serum-free medium (SFM) and a serum-enhanced culture medium. DESIGN: Chondrocytes were recovered from enzymatically digested human septal cartilage and were plated for monolayer culture in a newly developed medium. The medium included Dulbecco modified Eagle medium mixed 1:1 with Ham F12 medium and a supplement of known amounts of 2 growth factors-bFGF (100 ng/mL) and IGF-I (100 ng/mL)-used in combination and separately. RESULTS: The combination of IGF-I and bFGF enhanced chondrocyte growth and maintained a high degree of viability in SFM and 10% fetal calf serum. After an initial lag, the SFM, augmented with both growth factors, produced a comparable number of viable cells (4.25+/-0.31 x 10(4)) to that of the medium with 10% fetal calf serum (4.64+/-0.35 x 10(4)) by the seventh day of the experiment. Combined with the 2 growth factors, 10% fetal calf serum provided the greatest proliferation by the end of the experiment. However, the overall mean cell counts for the IGF-I- and bFGF-enhanced SFM were not statistically different. CONCLUSIONS: The combination of IGF-I and bFGF in a serum-free and a serum-supplemented environment supports the growth and viability of human septal chondrocytes in short-term culture. In an SFM, the results obtained approximate those produced in a medium enhanced with 10% fetal calf serum. PMID- 9865755 TI - Monocyte chemotactic and activating factor/monocyte chemoattractant protein-1 mediated histamine release from human nasal mucosa. AB - OBJECTIVES: To demonstrate the existence and localization of monocyte chemotactic and activating factor or monocyte chemoattractant protein-1 (MCAF/MCP-1) in human nasal mucosa and to verify its activity as a histamine-releasing factor. DESIGN: Detection of MCAF/MCP-1 in culture supernatants of nasal mucosa using Western blot analysis and assay of histamine release from basophils induced by these culture supernatants. Detection of MCAF/MCP-1 expression in nasal mucosa of patients with perennial allergic rhinitis using immunohistochemistry. PATIENTS: Twenty-one patients with house dust mite allergy, 7 nonallergic patients, and 5 patients with chronic inflammatory sinusitis participated in the study. All the allergic patients had positive test results for mite nasal allergy, detected by a clinical history, a nasal provocation test, and determination of specific mite IgE antibodies by a radioallergosorbent test. RESULTS: In Western blot analysis of supernatants of explant culture of human nasal mucosa, the band corresponding to approximately 13 to 15 kd was observed. This band was considered to be MCAF/MCP-1. These supernatants induced histamine release from basophils (approximately 3%-5% in net histamine release), and anti-MCAF/MCP-1 antibody inhibited this histamine-releasing activity. Immunoreactivity of MCAF/MCP-1 was observed in the nasal submucosa but not in the epithelium. Immunoreactive cells of MCAF/MCP-1 were also stained with the antibody, which recognizes monocytes and macrophages. CONCLUSIONS: These results suggest that MCAF/MCP-1, which is produced constantly by monocytes and macrophages and is stored in human nasal mucosa, possibly participates in the protracted histamine release from basophils and in the pathogenesis of perennial allergic rhinitis. PMID- 9865756 TI - Serologic study of the working mechanisms of immunotherapy for children with perennial allergic rhinitis. AB - BACKGROUND: Recent double-blind placebo-controlled trials have clearly shown the efficacy of immunotherapy for perennial allergic rhinitis. However, the exact working mechanisms related to the clinical effect of immunotherapy remain unclear. OBJECTIVES: To monitor the changes over time in immunologic parameters in children who received immunotherapy for perennial allergic rhinitis, and to elucidate the working mechanisms of immunotherapy related to its clinical efficacy. DESIGN: Nineteen children with perennial allergic rhinitis due to Dermatophagoides farinae enrolled in this prospective open study. Venous blood was collected to determine levels of specific IgE, specific IgG4, soluble interleukin 2 receptor, interleukin 4, soluble intercellular adhesion molecule 1, and soluble vascular cell adhesion molecule 1 at enrollment and 1, 2, 3, 5, and 10 years after enrollment. RESULTS: Immunotherapy affected serum levels of specific IgE, specific IgG4, soluble interleukin 2 receptor, interleukin 4, and soluble intercellular adhesion molecule 1, but not soluble vascular cell adhesion molecule 1. The rates of increase of levels of specific IgG4 and the rates of decrease of levels of soluble interleukin 2 receptor were correlated with the rates of decrease of symptom scores during the first 3 years of treatment, but not after 5 years. The rates of decrease in levels of soluble intercellular adhesion molecule 1 were correlated with the rates of decrease in symptom scores at 3 and 5 years after the beginning of the course of immunotherapy. The rates of decrease in levels of specific IgE and interleukin 4 were correlated with the rates of decrease in symptom scores after 5 and 10 years of treatment, but not during the first 3 years. CONCLUSION: Each modulation in levels of specific IgE, specific IgG4, soluble interleukin 2 receptor, interleukin 4, and soluble intercellular adhesion molecule 1 contributed to the clinical effect of immunotherapy in particular phases of treatment for children with perennial allergic rhinitis. PMID- 9865757 TI - Microbiologic characteristics of persistent otitis media. AB - OBJECTIVE: To identify the pathogens isolated from children with acute otitis media who did not respond to antimicrobial drug therapy. METHODS: Retrospective analysis of cultures obtained by tympanocentesis from 46 children. RESULTS: Organisms were recovered from 34 children (74%), and 43 isolates were recovered from these individuals. The organisms were Streptococcus pneumoniae (16 isolates), Haemophilus influenzae non-type b (12 isolates), Moraxella catarrhalis (5 isolates), Streptococcus pyogenes (5 isolates), Staphylococcus aureus (3 isolates), and Peptostreptococcus species (2 isolates). Resistance to the antimicrobial agent used was found in 27 (63%) of 43 isolates found in 22 patients (48%). Of patients who did not respond to amoxicillin therapy, H influenzae predominated. Streptococcus pneumoniae was recovered from 5 (56%) of 9 of those who did not respond to trimethoprim and sulfamethoxazole therapy, 4 (44%) of 9 patients after azithromycin therapy, 3 (25%) of 12 patients after amoxicillin therapy, and 2 (40%) of 5 patients after cefixime therapy. Streptococcus pyogenes was recovered from 2 (40%) of 5 patients after trimethoprim and sulfamethoxazole therapy and from 2 (40%) of 5 patients after cefixime therapy. CONCLUSIONS: The data illustrate the relation between resistance to antimicrobial drug therapy and failure of patients with otitis media to improve. They also highlight the importance of diagnostic tympanocentesis in establishing the presence of resistant microorganisms. PMID- 9865758 TI - Otogenic intracranial abscess: concurrent craniotomy and mastoidectomy--changing trends in a developing country. AB - OBJECTIVES: To evaluate (1) the clinical profile, treatment, and outcome of adult and pediatric patients presenting with intracranial abscess of otogenic origin and (2) the advantages of concurrent craniotomy and mastoidectomy. DESIGN: A prospective case series. SETTING: An academic tertiary referral center in India. PATIENTS: Thirty-six patients clinically diagnosed as having intracranial abscess that was secondary to suppurative otitis media and confirmed by computed tomographic scanning. INTERVENTION: Concurrent craniotomy and mastoidectomy. RESULTS: Children were more commonly affected than adults, and there was a male preponderance. All patients had cholesteatoma at surgery, although one third of the children had only granulation tissue on otoscopy. More than two thirds of the patients in both the groups presented with more than one intracranial complication and definitive surgical intervention was done later than 24 hours. Meningitis was the most frequent intracranial complication, followed by cerebellar abscess. There was no significant intraoperative or postoperative morbidity, mortality, recurrence of intracranial complications, or residual neurological deficits. Three children (14%) showed evidence of recidivism cholesteatoma requiring revision surgery. CONCLUSIONS: In suppurative otitis media with intracranial complications, it is accepted practice to treat the neurosurgical complication first, followed by mastoidectomy at a later date after the patient has been stabilized. Craniotomy with concurrent mastoidectomy is not only safe, but it also removes the source of infection at the same time the complications are being treated, thus avoiding reinfection while the patient is awaiting the ear surgery. In addition, the treatment is completed with a single, shorter hospital stay, which is more economical for the patient. PMID- 9865759 TI - Expression of the transforming growth factor beta isoforms in inflammatory cells of nasal polyps. AB - OBJECTIVE: To determine the expression and the potential role of transforming growth factor beta (TGF-beta) in nasal polyposis. DESIGN: Comparison of TGF-beta expression between normal and inflammatory nasal mucosa and polyps; in inflammatory nasal polyps, characterization of the TGF-beta isoforms expression and their potential location in macrophages and eosinophils. SETTING: Patients and samples were selected at the Hopital Intercommunal, Creteil, France, and immunohistochemistry and immunoblots were performed at the Institut National de la Sante et de la Recherche Medicale U296 (Universite Paris XII, France). SUBJECTS: Nasal polyps and nasal mucosa were sampled in 21 patients during ethmoidectomy, and muscosa was sampled in 6 healthy patients during rhinoplasty. METHODS: Immunohistochemistry and Western blot analysis were performed using specific antibodies to TGF-beta1-3, TGF-beta1, TGF-beta2, and TGF-beta3 isoforms. Double labeling was also performed using anti-TGF-beta1 antibody together with macrophages or eosinophil-specific antibodies. RESULTS: The expression of TGF beta(1-3) was significantly higher in inflammatory nasal polyps than in inflammatory nasal mucosa and higher in inflammatory nasal mucosa than in nasal mucosa from healthy patients. Transforming growth factor beta1 was the main isoform detected in inflammatory nasal polyps, and it was present in numerous macrophages and in some eosinophils. CONCLUSIONS: Transforming growth factor beta, mainly TGF-beta1, is strongly expressed in inflammatory nasal mucosa, where it could be produced by macrophages and eosinophils. Transforming growth factor beta could induce epithelium and connective tissue modifications and therefore be involved in the pathogenesis of nasal polyposis. PMID- 9865760 TI - Perinatal management of unanticipated congenital laryngeal atresia. AB - Unless ventilation is achieved within minutes of delivery, patients with congenital laryngeal atresia will not survive. There are 2 settings in which survival is more likely: a tracheotomy may be immediately performed in the delivery room, or a communication may exist between the airway and the pharynx, allowing for air exchange. In the latter case, there are no characteristic findings on prenatal sonography to suggest the diagnosis and to ensure that preparations for immediate tracheotomy are made. We describe a neonate with unanticipated laryngeal atresia and a high tracheoesophageal fistula. Ventilation was maintained first by face mask and then by esophageal intubation until a tracheotomy could be performed. This report provides detailed photodocumentation of the anomaly, discusses the mechanism of air exchange, reviews the relevant embryological development, and outlines a protocol for perinatal management of unanticipated laryngeal atresia. PMID- 9865761 TI - Hypopharyngeal stenosis and dysphagia complicating toxic epidermal necrolysis. AB - Toxic epidermal necrolysis is a severe dermatologic disorder clinically characterized by the acute onset of erythema and tenderness of the skin. Destruction of the epidermal barrier results in significant morbidity and mortality. Large erosions of mucous membrane, including the mouth and oral mucosa, are typical of toxic epidermal necrolysis. After ingesting naproxen sodium (Aleve) and aspirin, a previously healthy 43-year-old woman developed toxic epidermal necrolysis that resulted in hypopharyngeal stenosis complicated by dysphagia and recurrent aspiration. PMID- 9865762 TI - In vivo correlation of neutrophil receptor expression, ischemia-reperfusion injury, and selective 5-lipoxygenase inhibition in guinea pigs. AB - OBJECTIVE: To determine whether selective 5-lipoxygenase (5-LO) inhibition decreases expression of adhesion molecules (beta2 integrins) on systemic neutrophils, decreases neutrophil infiltration in ischemic flap tissue, and improves flap survival. DESIGN: A randomized, controlled study of 91 adult female Hartley guinea pigs divided into 3 survival groups, 4 neutrophil assay groups, 1 sham group, and 1 control group. Ischemia of varying duration and reperfusion was induced in island flank skin flaps. The treated groups received zileuton, a 5-LO inhibitor, orally during flap ischemia. After reperfusion, systemic neutrophil receptor expression, neutrophil infiltration, and flap survival were measured. Surface receptor molecules on neutrophils from whole blood samples obtained via transcardiac puncture were analyzed using monoclonal antibodies and cell associated fluorescence. Neutrophil infiltration into a distal 1 cm2 of flap tissue was assessed using myeloperoxidase antibodies. Flap survival was determined within 7 days of surgery. RESULTS: Untreated flaps with 10 hours of ischemia underwent total necrosis. Treated 2- and 10-hour ischemic flaps survived intact. A significant main effect of the drug treatment was detected using analysis of variance (P<.001). Neutrophil receptor detection in the untreated groups undergoing 2 and 10 hours of ischemia was significantly increased compared with that in the treated groups with the same ischemia times. Skin neutrophil infiltration was significantly decreased in the treated groups. CONCLUSIONS: Systemic administration of a 5-LO inhibitor is effective in reducing ischemia reperfusion injury in flap tissue. Our data indicate that there is a significant reduction in neutrophil receptor expression with administration of 5-LO, reducing the priming of systemic neutrophils from circulating cytokines. PMID- 9865763 TI - Osteopontin and bone sialoprotein distribution at the bone graft recipient site. AB - BACKGROUND: The area in close proximity to a bone graft is subject to marked remodeling activity, which may dramatically affect graft survival. OBJECTIVE: To specifically analyze the effects at the recipient bed-onlay graft interface. DESIGN: In 22 adult Lewis rats, bicortical grafts were positioned below the temporal muscle and subperiosteally over the parietal bone. The recipient bone was left intact or ground to remove the external cortical layer, thereby exposing the graft to the osteopotent cells of the bone marrow. The rats were killed after 4 or 20 weeks. The outcome was assessed by routine histologic examination and immunohistochemical labeling for 2 bone matrix proteins, osteopontin and bone sialoprotein, which are involved in bone resorption and formation, respectively. RESULTS: Placement of the grafts submuscularly or grinding of the outer cortical layer of the host bed increased recipient site resorption. Resorptive activity (labeling) was concentrated to a subzone below the surface of the recipient bone; neither the graft surface nor the interface soft tissues were labeled. CONCLUSION: The successive loss of skeletal contour after bone grafting, in many cases, may largely result from recipient site failure rather than graft size reduction. PMID- 9865764 TI - Second-look endoscopic sinus surgery. PMID- 9865765 TI - Can unilateral hearing loss be a handicap in learning? PMID- 9865766 TI - Lower reconstruction and restoration of oral competence with dynamic palmaris longus vascularized sling. PMID- 9865767 TI - Pathologic quiz case 1: Nasopharyngeal teratoma. PMID- 9865768 TI - Pathologic quiz case 2: Langerhans cell histiocytosis (LCH) of the temporal bone. PMID- 9865769 TI - Selection of materials for orbital floor reconstruction. PMID- 9865770 TI - Bone grafting for defects of the orbital floor. PMID- 9865771 TI - Surgery of traumatic defects of the orbital floor. PMID- 9865772 TI - Spatial audiometry: detection of spondaic words in noise. AB - It is important to select appropriate stimuli and test conditions for developing standardized spatial audiometric tests. In three experiments, binaural detection thresholds (BDTs) for a target signal, located at either 0, 45, 90, 135, 180, 225, 270, or 315 degrees azimuth, were measured in the presence of a masker positioned at one of these eight locations. Target signals included spondaic words from the CID W-1 list. The masker was speech spectrum noise (SSN) or multitalker noise (MTN) presented at a constant level (65 dBA). Bekesy tracking was used to measure BDTs in listeners with normal hearing. Results indicate that BDTs are significantly influenced by the (a) angular separation between the target and noise source and (b) choice of spondaic words used as target stimuli. BDTs for various spondaic words differed as much as 13 dB for a given angular separation. BDTs measured in SSN and MTN for otherwise identical test conditions differed less than 3 dB. A single spondaic word appears to be appropriate for spatial audiometric tests of detection. Nonsignificant differences between masked BDTs obtained for SSN and MTN noises indicate that for spatial detection, the masking effects of these noises are comparable. These results indicate that the development of a clinical test of spatial detection should include the use of SSN and a single spondaic word, with detection being measured for a set of four or five signal and noise source configurations. PMID- 9865773 TI - Quantifying air-conducted acoustic radiation from the bone-conduction vibrator. AB - Sound pressure levels in the external auditory canals of 50 subjects were measured at 2000 and 4000 Hz with a bone-conduction vibrator on the forehead, the mastoid ipsilateral to the probe microphone, and the mastoid contralateral to the probe microphone. A plug was placed in the external auditory canal to minimize sound pressure levels in the external auditory canal produced by the osseotympanic mode of bone conduction. Results suggest that clinically significant false air-bone gaps (greater than 10 dB) due to acoustic radiation into the concha from the bone-conduction vibrator are most likely to occur at 4000 Hz when the bone-conduction vibrator is placed on the mastoid of the test ear. To minimize the possible confounding effects of acoustic radiation, the bone conduction vibrator may be placed on the forehead or the mastoid contralateral to the test ear while masking the nontest ear. PMID- 9865774 TI - Development of suprathreshold word recognition test for Russian-speaking patients. AB - This paper describes the development and normalization of a computerized word recognition test for speakers of the Russian language: the Russian Picture Identification Task (RPIT). The test uses a picture-pointing response, in which a patient selects, on a computer screen, a picture corresponding to a target word from a foil of four rhyming alternatives. The auditory portion of the test is recorded digitally and is recalled from a clinician's computer by clicking a mouse on the English translation of the target word. Thus, an English-speaking clinician can assess reliably word recognition abilities of Russian-speaking patients-a task that has become important in view of the recent surge of immigration from the former Soviet Union. Normative performance of normal-hearing native speakers of Russian was obtained in oral response conditions and picture pointing conditions intended for clinical use. Psychometric functions provide norms for clinical decision making. Error patterns, individual confusions, and cognitive aspects of the experiment are discussed. PMID- 9865775 TI - Real-ear characteristics of the ALGO2 acoustic transducer assembly. AB - Clinical reports regarding the use of the ALGO2 neonatal automated auditory brainstem response (AABR) screening device have been concerned only with issues of sensitivity and specificity. This study was undertaken to evaluate how couplers and their placement affect the ALGO2 click spectral properties. Although not recommended by the manufacturer, substituting less expensive, uncalibrated couplers for the standard earphone may be a tempting alternative in clinical practice. Click spectra of an ALGO2 were analyzed with a real-ear system so that comparisons could be made between probe tips and the standard coupler. In addition, ALGO2 screening data from neonates using the standard earphone and a substitute probe tip were compared. The results are in agreement with the manufacturer's specifications when the ALGO2 is used as prescribed. However, with probe tip couplers, SPL values were markedly increased, particularly as insertion depth of the probes varied. The neonatal AABR results show that coupler type and placement can also produce inaccurate screening evaluations and erroneous conclusions. PMID- 9865776 TI - Telephone communication with in-the-ear hearing aids using acoustic and electromagnetic coupling. AB - The purpose of this study was to determine whether telephone communication was improved with electromagnetic coupling for in-the-ear hearing aid users with mild to moderate hearing losses. Each user adjusted his hearing aid gain to the level he determined to be best for telephone communication with each coupling method. Under these conditions, average word recognition scores in quiet and average high frequency output levels measured at the tympanic membrane were not different for each coupling method. The ability to tolerate background noise was also studied. Background noise tolerance was not different for the two coupling methods. However, significantly greater levels of background noise were tolerated when telephone side-tone feedback was eliminated regardless of coupling method. PMID- 9865777 TI - Effect of gender on the mismatch negativity auditory evoked potential. AB - Mismatch negativity (MMN) was measured on normal-hearing young adult women and men to determine the effect of gender on this auditory evoked potential (AEP). In the experimental condition, recordings were obtained for 1000-Hz tone bursts presented at 75 dB nHL (standard stimuli) and 60 dB nHL (deviant stimuli). AEPs also were obtained in a control condition in which all stimuli were presented at 60 dB nHL; however, 15 percent of these responses were averaged to represent a response analogous to the experimental deviant response. The MMN was derived by subtracting the analogous control waveform from the experimental deviant waveform. Measures of peak-to-peak amplitude, peak latency, and area-under-the curve were obtained for each derived waveform. Analysis of these data indicated no significant gender differences in peak latency of the MMN response. However, peak-to-peak amplitude and area-under-the-curve were significantly larger for women than for men. PMID- 9865778 TI - Professional education and assessment practices in central auditory processing. AB - A 17-item questionnaire probing professional preparation and current practices in central auditory assessment was mailed to 500 audiologists selected randomly from the membership directory of the American Academy of Audiology. Data from 183 respondents, representing a 37 percent response rate, were analyzed. The majority of respondents reported minimal academic and clinical preparation in assessment of the central auditory nervous system. Eighty percent of respondents had not taken any graduate course explicitly dedicated to central auditory processing. However, 80 percent had taken at least one basic science course in central audition and 83 percent reported having taken at least one graduate course that included some coverage of central auditory processing and/or the central auditory nervous system. A mean of 3 clinical clock hours accrued in this area was reported. Not surprisingly, 78 percent reported a satisfaction rating of < or =50 percent relative to the graduate education they received in this area and only 41 percent reported providing central auditory assessment. Comparisons with prior surveys show substantial change in the preferred test battery. Most notable is the pivotal role of physiologic measures, with the acoustic reflex and auditory brainstem response listed along with the SCAN as the three most frequently used assessment tests/procedures. Overall, the results suggest a need for improvement in professional preparation in evaluation of central auditory function. PMID- 9865779 TI - Sound localization in the median sagittal plane by listeners with presbyacusis. AB - In Experiment 1, a group of listeners with substantial hearing loss due to presbyacusis and a group of listeners with normal hearing were given three localization tests: a frontal plane test in which they judged whether sounds came from the left, overhead, or the right; a sagittal plane test in which they judged whether sounds came from directly in front, overhead, or behind; and an elevation test in which they judged the vertical position of sounds coming from in front. The two groups performed similarly on the frontal plane test, which chiefly depended upon their ability to use binaural localization cues. They performed differently on the sagittal plane and elevation tests, for which the predominant localization cues were spectral. The listeners with presbyacusis were substantially less accurate than those with normal hearing in both of these instances. They had particular difficulty judging source elevation, rarely scoring much above chance. Follow-up testing of a group of subjects in the early stages of presbyacusis showed localization performance that was intermediate to the other two groups, but far more like that of the normal-hearing listeners. In Experiment 2, additional tests were run with the following conditions designed to encourage improved performance by listeners with presbyacusic hearing loss: (1) filtering of stimuli to preclude masking of more informative high-frequency components by low frequencies; (2) simplification of the elevation test and greater spatial separation of its loudspeaker sources; and (3) use of hearing aids. Conditions 1 and 2 had no appreciable effect on performance; condition 3 significantly improved presbyacusic listeners' ability to localize in the sagittal plane, particularly when sounds came from the front. PMID- 9865780 TI - Mapping of members of the low-copy-number repetitive DNA sequence family chAB4 within the p arms of human acrocentric chromosomes: characterization of Robertsonian translocations. AB - Members of the long-range, low-copy-number repetitive DNA sequence family chAB4 are located on nine different human chromosome pairs and the Y chromosome, i.e. on the short arms of all the acrocentrics. To localize the chAB4 sequences more precisely on the acrocentrics, chAB4-specific probes together with rDNA and a number of satellite sequences were hybridized to metaphase chromosomes of normal probands and of carriers of Robertsonian translocations of the frequent types rob(13q14q) and rob(14q21q). The results demonstrate that chAB4 is located on both sides of the rDNA on all the acrocentrics; the exact location, however, may be chromosome specific. Chromosome 22, most probably, is the only chromosome where chAB4 is found in the direct neighbourhood of the centromere. Fluorescence in situ hybridization analyses of metaphase chromosomes of carriers of rob(21q22q) revealed breakpoint diversity for this rare type of Robertsonian translocation chromosome. A direct involvement of chAB4 sequences in recombination processes leading to the Robertsonian translocations analysed in this study can be excluded. PMID- 9865781 TI - Chromosome banding studies in the Bateleur (Terathopius ecaudatus, Aves, Accipitridae). AB - The karyotype of the Bateleur (Terathopius ecaudatus) was studied with conventional and Ag-NOR staining, and using GTG and CBG banding. The karyotype organization is typically accipitrid, with satellites and few microchromosomes, close to the karyotypes of true vultures. PMID- 9865783 TI - Mapping chromosomal homologies between humans and two langurs (Semnopithecus francoisi and S. phayrei) by chromosome painting. AB - Chromosomal homologies were established between human and two Chinese langurs (Semnopithecus francoisi, 2n = 44, and S. phayrei, 2n = 44) by chromosome painting with chromosome-specific DNA probes of all human chromosomes except the Y. Both langur species showed identical hybridization patterns in addition to similar G-banding patterns. In total, 23 human chromosome-specific probes detected 30 homologous chromosome segments in a haploid langur genome. Except for human chromosomes 1, 2, 6, 16 and 19 probes, which each gave signals on two non homologous langur chromosomes respectively, all other probes each hybridized to a single chromosome. The results indicate a high degree of conservation of chromosomal synteny between human and these two Chinese langurs. The human chromosome 2 probe painted the entire euchromatic regions of langur chromosomes 14 and 19. Human chromosome 1 probe hybridized to three regions on langur autosomes, one region on langur chromosome 4 and two regions on langur chromosome 5. Human 19 probe hybridized on the same pattern to one region on chromosome 4 and to two regions on langur chromosome 5, where it alternated with the human chromosome 1 probe. Human 6 and 16 probes both hybridized to one region on each of the two langur autosomes 15 and 18. Only two langur chromosomes (12 and 21) were each labelled by probes specific for two whole human chromosomes (14 and 15 and 21 and 22 respectively). Comparison of the hybridization patterns of human painting probes on these two langurs with the data on other Old World primates suggests that reciprocal and Robertsonian translocations as will as inversions could have occurred since the divergence of human and the langurs from a common ancestor. This comparison also indicates that Asian colobines are karyotypically more closely related to each other that to African colobines. PMID- 9865782 TI - DNA methylation patterns of Melandrium album chromosomes. AB - Melandrium album (syn. Silene latifolia) belongs to dioecious plant species possessing heteromorphic sex chromosomes, X and Y. Our previous experiments using in situ nick translation and replication kinetics analysis indicated structural and functional differences between the two X chromosomes in homogametic female cells. Here we show DNA methylation patterns of M. album root tip chromosomes using the indirect immunofluorescence approach with a monoclonal antibody raised against 5-methylcytosine (5-mC). In male cells, a more intensive 5-mC labelling on the shorter arm of the only X chromosome was observed in comparison with the longer X arm. A global hypermethylation of the male Y chromosome was not found, which indicates its prevalent euchromatic character. In female cells, the specific 5-mC pattern of the X chromosome was found on a single X chromosome, whereas the other X displayed an overall higher level of 5-mC labelling. Application of a hypomethylating drug, 5-azacytidine (5-azaC), during seed germination led to a deletion of any specific differences in the 5-mC distribution between the two X chromosomes. Confocal laser scanning microscopy analysis of DNA methylation in interphase nuclei showed hypermethylated domains that were efficiently decondensed and hypomethylated by 5-azaC treatment. The presented data show reproducible differences in the DNA methylation patterns between the two X chromosomes in M. album female somatic cells, which indicate their distinct transcriptional activities as a possible consequence of the negative dosage compensation of X-linked genes. PMID- 9865784 TI - Heterogeneity and meiotic behaviour of B and sex chromosomes, banding patterns and localization of (TTAGGG)n sequences by fluorescence in situ hybridization in the neotropical water rat Nectomys (Rodentia, Cricetidae). AB - A cytogenetic study using fluorescence in situ hybridization (FISH) of telomere probes, CBG, GTG and RBG banding patterns and synaptonemal complex data was carried out in 41 specimens of Nectomys from three Brazilian states: Pernambuco, Mato Grosso and Sao Paulo. The specimens presented 2n = 52, 53, 56 and 57, and the differences in chromosome number were due to the presence of three different supernumeraries and also to the occurrence of tandem fusions. The tandem fusions involved chromosome pairs 3 + 11 and 5 + 24 from karyotype with 2n = 56 that originated pairs 1 and 4 in specimens with 2n = 52. Sex chromosome polymorphism was also detected, and the X presented three different morphologies, which could be explained by heterochromatin addition/deletion. FISH results revealed interstitial telomeric bands (ITBs) in a submetacentric B, but no ITB was detected in the chromosomes originated by tandem fusion. The supernumeraries presented a remarkable heterogeneity of size and morphology, constitutive heterochromatin pattern and localization of telomeric sequences. Synaptonemal complex by light and electron microscopy showed the supernumerary as an autopaired univalent. The sex chromosome pairing in meiotic cells involved the heterochromatic short arm of the X chromosome and the short arm of the Y chromosome. PMID- 9865785 TI - Chromosomal localization and heterochromatin association of ribosomal RNA gene loci and silver-stained nucleolar organizer regions in salmonid fishes. AB - Ribosomal RNA gene (rDNA) loci, including those of nucleolus-forming 18S, 5.8S and 28S (major) and non-nucleolus-forming 5S (minor) rDNA, were assigned using fluorescence in situ hybridization (FISH) to the embryonic chromosomes of rainbow trout (Oncorhynchus mykiss), masu salmon (O. masou), brook trout (Salvelinus fontinalis) and Japanese huchen (Hucho perryi). In these species, the minor rDNA loci were located basically on 2-4 chromosome pairs, whereas the major rDNA loci were found essentially on one chromosome pair, except for the brook trout. Its major rDNA loci were dispersed on about half of the chromosome complement, showing a considerable interindividual variation in the number and location. The major and minor rDNA loci were separated onto different chromosomes in the examined species, except for the rainbow trout, in which one chromosome pair had tandemly aligned minor and major rDNA loci. Chromosome regions containing both kinds of rDNA loci in each species were found to be stained with C-banding, showing an association of these loci with heterochromatin. Comparison of the assigned major rDNA loci and sequentially detected silver (Ag)-stained nucleolar organizer regions (AgNORs) in all the species revealed a considerable polymorphism in the number and size of AgNORs among or within those loci, suggesting a possible inter- or intralocus inactivation of the major rDNAs. PMID- 9865786 TI - Condensation pattern (CP) analysis of plant chromosomes by an improved chromosome image analysing system, CHIAS III. AB - The third-generation chromosome image analysing system (CHIAS III) was developed by using an ordinary personal computer with public domain imaging software, the NIH Image. An uneven condensation pattern (CP) appearing on small sugarcane chromosomes at the mitotic prometaphase stage was analysed and a quantitative chromosome map or idiogram was developed by using CHIAS III. The analysis consists of three major steps as follows: (1) manipulation of original chromosome images and measurement of CP or numerical data of the CP; (2) calculation of an averaged CP for homologous chromosomes; and (3) development of an idiogram based on the averaged CP. The CP has been analysed quantitatively by using the first- and the second-generation chromosome image analysing systems with hard- and software specific to the systems. The data obtained using CHIAS III are consistent with those obtained with the chromosome image analysing systems previously developed. PMID- 9865787 TI - Fibre-fluorescence in situ hybridization unravels apparently seven DAZ genes or pseudogenes clustered within a Y-chromosome region frequently deleted in azoospermic males. AB - Using the technique of 'fibre-FISH' (fluorescence in situ hybridization), we describe the direct visualization of seven longer DAZ signal stretches and in addition a maximum of four isolated single DAZ signals on Y-chromatin fibres of four different individuals. These seven longer DAZ signal stretches may represent seven DAZ genes or pseudogenes, whereas the single DAZ signals may represent truncated DAZ genes. PMID- 9865788 TI - Interspecies comparative genome hybridization and interspecies representational difference analysis reveal gross DNA differences between humans and great apes. AB - Comparative chromosome G-/R-banding, comparative gene mapping and chromosome painting techniques have demonstrated that only few chromosomal rearrangements occurred during great ape and human evolution. Interspecies comparative genome hybridization (CGH), used here in this study, between human, gorilla and pygmy chimpanzee revealed species-specific regions in all three species. In contrast to the human, a far more complex distribution of species-specific blocks was detected with CGH in gorilla and pygmy chimpanzee. Most of these blocks coincide with already described heterochromatic regions on gorilla and chimpanzee chromosomes. Representational difference analysis (RDA) was used to subtract the complex genome of gorilla against human in order to enrich gorilla-specific DNA sequences. Gorilla-specific clones isolated with this technique revealed a 32-bp repeat unit. These clones were mapped by fluorescence in situ hybridization (FISH) to the telomeric regions of gorilla chromosomes that had been shown by interspecies CGH to contain species-specific sequences. PMID- 9865789 TI - Inhibition of condensation in human chromosomes induced by the thymidine analogue 5-iododeoxyuridine. AB - Human lymphocyte cultures were treated with iododeoxyuridine. This 5-halogenated thymidine analogue induces distinct undercondensations of the heterochromatin of human chromosome 9. The condensation of the other heterochromatic regions on chromosomes 1, 15, 16 and Y is also inhibited, but to a lesser extent. Optimum cell culture conditions required for the induction of undercondensations were determined. Up to 90% of mitotic cells reveal chromosome 9 heterochromatin decondensations when the substance is present in quantities of 1 x 10(-4) M during the last 7 h before cell fixation. In addition, examples of the usefulness of 5-IUDR in the analysis of chromosome aberrations involving chromosome 9 are presented. The interaction between 5-IUDR and chromosomal DNA, the modification of chromosomal proteins and factors inducing chromosomal decondensations are discussed. PMID- 9865790 TI - Fibre-fluorescence in situ hybridization directly performed from fresh biological samples: novel perspectives for genetic diagnosis. PMID- 9865791 TI - Aneuploidy in bovine oocytes matured in vitro. PMID- 9865792 TI - Intraepidermal nerve fiber assessment: a new window on peripheral neuropathy. PMID- 9865793 TI - Molecular immunology and genetics of inflammatory muscle diseases. AB - Polymyositis, dermatomyositis, and inclusion body myositis, although immunopathologically distinct, share 3 dominant histological features: inflammation, fibrosis, and loss of muscle fibers. Progress in molecular immunology and immunogenetics has enhanced our understanding of these cellular processes. Based on the T-cell receptor gene rearrangement, the autoinvasive CD8+ T cells in polymyositis and inclusion body myositis, but not dermatomyositis, are specifically selected and clonally expanded in situ by heretofore unknown muscle specific autoantigens. The messenger RNA of cytokines is variably expressed, except for a persistent up-regulation of interleukin 1beta in inclusion body myositis and transforming growth factor beta in dermatomyositis. In inclusion body myositis, the interleukin 1, secreted by the chronically activated endomysial inflammatory cells, may participate in the formation of amyloid because it up-regulates beta-amyloid precursor protein (beta-APP) gene expression and beta-APP promoter and colocalizes with beta-APP within the vacuolated muscle fibers. In dermatomyositis, transforming growth factor beta is overexpressed in the perimysial connective tissue but is down-regulated after successful immunotherapy and reduction of inflammation and fibrosis. The degenerating muscle fibers express several antiapoptotic molecules, such as Bcl-2, and resist apoptosis-mediated cell death. In myositis, several of the identified molecules and adhesion receptors play a role in the process of inflammation, fibrosis, and muscle fiber loss, and could be targets for the design of semispecific therapeutic interventions. PMID- 9865795 TI - The G209A mutation in the alpha-synuclein gene is not detected in familial cases of Parkinson disease in non-Greek and/or Italian populations. AB - OBJECTIVE: To determine whether the G-to-A substitution at nucleotide 209 (G209A) mutation in the alpha-synuclein gene is responsible for familial Parkinson disease (PD) in the US population. DESIGN: Polymerase chain reaction-based DNA analysis of consecutive patients with PD and family history of PD. SETTING: A university-affiliated movement disorder clinic and a Veterans Affairs clinical research laboratory. PATIENTS: Forty-four patients with PD and family history of PD and 29 patients with sporadic PD, all with no known Greek and/or Italian background. RESULTS: None of the DNA samples showed the G209A mutation. CONCLUSION: The G209A mutation is rare in US patients with familial PD. PMID- 9865794 TI - Epidermal nerve fiber density: normative reference range and diagnostic efficiency. AB - BACKGROUND: The sensitivity of neuron-specific antibodies permit the identification of the small unmyelinated nerve fibers within the skin. OBJECTIVES: To develop a reference range of epidermal nerve fiber density in humans, and to evaluate their diagnostic efficiency for sensory neuropathies. METHODS: Ninety-eight normal controls (age range, 13-82 years) were examined with both directed neurologic examinations and quantitative sensory testing. The diagnostic utility was examined in 20 patients with sensory neuropathies. Each subject had 2 punch biopsies performed at each site in the thigh and distal part of the leg (total of 392 biopsies). After formalin fixation, 50-microm-thick free floating sections were stained with a polyclonal antibody to neuron-specific ubiquitin hydrolase, anti-protein gene product 9.5. We enumerated intraepidermal nerve fibers per millimeter to derive a "linear density." The linear density technique was validated against a stereological technique that used the fractionator to measure the total length of intraepidermal nerve fibers per 3-mm punch. RESULTS: The biopsy technique was well tolerated, with no notable complications. The linear density quantitation was rapid and had high intraobserver and interobserver reliability. We determined that the density of intraepidermal fibers in normal controls was 21.1+/-10.4 per millimeter (mean +/- SD) in the thigh (fifth percentile, 5.2 per millimeter), and was 13.8+/-6.7 per millimeter at the distal part of the leg (fifth percentile, 3.8 per millimeter). Significantly higher intraepidermal fiber densities were seen in the youngest group (P = .004), and we observed no significant effect of race, sex, height, or weight. The density at the thigh was significantly correlated with that at the distal part of the leg (P = .01) and was consistently higher by about 60%, a reflection of the normal proximal-distal gradient. The results obtained with stereology and the linear density correlated significantly (P=.001), providing internal validation for the technique. Epidermal nerve fiber density was significantly reduced (P = .001) in patients with sensory neuropathies. With a cutoff derived from the fifth percentile of the normative range for the distal part of the leg, the technique had a positive predictive value of 75%, a negative predictive value of 90%, and a diagnostic efficiency of 88%. CONCLUSIONS: We have established a reference range for intraepidermal nerve fiber density in normal humans by means of a simple quantitation method based on enumeration of individual intraepidermal nerve fibers on vertical sections of punch skin biopsy specimens stained with the sensitive panaxonal marker anti-protein gene product 9.5. The utility of the density measurement was confirmed for sensory neuropathy with a diagnostic efficiency of 88%. Skin biopsies may be useful to assess the spatial distribution of involvement in peripheral nerve disease and the response to neurotrophic and other restorative therapies. PMID- 9865796 TI - Gamma knife radiosurgery for trigeminal neuralgia: results and expectations. AB - BACKGROUND: Trigeminal neuralgia is a disabling pain syndrome responsive to both medical and surgical therapies. Stereotactic radiosurgery using the gamma knife can be used to inactivate a specified volume in the brain by cross firing 201 photon beams. We evaluated pain relief and treatment morbidity after trigeminal neuralgia radiosurgery. METHODS: All evaluable patients (n = 106) had medically or surgically refractory trigeminal neuralgia. A single 4-mm isocenter of radiation was focused on the proximal trigeminal nerve just anterior to the pons. For follow-up an independent physician who was unaware of treatment parameters contacted all patients. RESULTS: After radiosurgery, 64 patients (60%) became free of pain and required no medical therapy (excellent result), 18 (17%) had a 50% to 90% reduction (good result) in pain severity or frequency (some still used medications), and 9 (9%) had slight improvement. At last follow-up (median, 18 months; range, 6-48 months), 77% of patients maintained significant relief (good plus excellent results). Only 6 (10%) of 64 patients who initially attained complete relief had some recurrent pain. Radiosurgery dose (70-90 Gy), age, surgical history, or facial sensory loss did not correlate with pain relief. Poorer results were found in patients with multiple sclerosis. Twelve patients developed new or increased facial paresthesias after radiosurgery (10%). No patient developed anesthesia dolorosa. There was no other procedural morbidity. CONCLUSIONS: Gamma knife radiosurgery is a minimally invasive technique to treat trigeminal neuralgia. It is associated with a low risk of facial paresthesias, an approximate 80% rate of significant pain relief, and a low recurrence rate in patients who initially attain complete relief. Longer-term evaluations are warranted. PMID- 9865797 TI - Anticoagulation therapy in pediatric patients with sinovenous thrombosis: a cohort study. AB - OBJECTIVE: To assess the use of anticoagulant therapy in a consecutive cohort study of children with sinovenous thrombosis (SVT). METHODS: A single institutional pilot study of anticoagulant therapy was conducted from January 1992 to December 1996 at the Hospital for Sick Children, Toronto, Ontario. Consecutive children with the diagnosis of SVT, made by computed tomography, magnetic resonance imaging (MRI), MRI with venography, ultrasonography, nuclear brain scanning, or conventional angiography were eligible for anticoagulant therapy. RADIOGRAPHIC EVALUATION: Most children underwent multiple radiographic tests for evaluation of the central nervous system. Of the 32 episodes of SVT, CT was performed in 30, MRI with or without venography in 26, ultrasonography in 11, and nuclear brain scanning in 5. The majority of the SVTs were located at the superior sagittal sinus (50%) and right lateral sinus complex (44%). RESULTS: There were 30 consecutive children with 32 episodes of SVT during the 5-year study (2 girls had recurrent SVT). The median age was 6.2 years (age range, 3 days to 18 years), and the sex of the patients was evenly distributed (15 girls and 15 boys). The primary associated clinical conditions consisted of systemic lupus erythematosus (n = 5), renal disease (n = 3), perinatal distress (n = 2), congenital heart disease (n = 1), cerebral arteriovenous malformation (n = 1), and neurosurgery for refractory seizures (n= 1). The remainder were previously healthy children older than 1 month (n = 10) and newborns (n = 7). Eight children were ineligible for anticoagulant therapy because of an associated intracranial hemorrhage (n = 6), a postoperative bleeding risk after neurosurgery (n = 1), or a prolonged delay from the diagnosis to the time of referral (n = 1). Ten children received standard heparin, and 12 children received low-molecular-weight heparin (LMWH) (enoxaparin sodium). Eighteen children were treated with oral anticoagulants for 3 months after initial heparin therapy, and 4 patients received LMWH for the entire course of treatment. There was no intracranial hemorrhage in the 12 patients treated with LMWH, but there was 1 case of clinically silent bleeding in the standard heparin group. CONCLUSIONS: The results of this pilot study suggest that anticoagulant therapy, in particular LMWH, is safe and may have a role in the treatment of children with SVT. A randomized controlled trial is warranted. PMID- 9865798 TI - Prothrombotic disorders in infants and children with cerebral thromboembolism. AB - BACKGROUND: To our knowledge, the contribution of prothrombotic conditions to cerebral thromboembolism has never been prospectively studied in a large series of pediatric patients. METHODS: The Hospital for Sick Children, Toronto, Ontario, established a program in January 1992 to diagnose and treat children (term newborn to 18 years old) with arterial ischemic stroke or sinovenous thrombosis. The routine evaluation for prothrombotic conditions included plasminogen, antithrombin, protein C, free protein S, activated protein C resistance, IgG and IgM anticardiolipin antibody, and lupus anticoagulant. We analyzed samples taken within 2 years of the event. We report results on patients seen from January 1, 1992, to January 1, 1997. RESULTS: Ninety-two patients (47 males and 45 females) entered the program during the study interval. Patients ranged from newborn to 18 years in age. Arterial ischemic stroke occurred in 78% of patients while sinovenous thrombosis occurred in 22%. All were tested for prothrombotic disorders. One or more abnormal results were present in 35 (38%) of the 92 patients. The majority (21/35) had multiple abnormal test results. The abnormal test results were anticardiolipin antibody (33%), plasminogen (9.5%), activated protein C resistance (9%), protein C (7%), antithrombin (12.5%), lupus anticoagulant (8%), and free protein S (11.5%). Male sex predicted the presence of prothrombotic abnormalities (relative risk, 1.7; 95% confidence interval, 1.2 2.5), but stroke type (relative risk, 0.8; 95% confidence interval, 0.7-1.1), age group, and presence of other risk factors did not predict abnormal testing. CONCLUSIONS: A significant proportion (38%) of children with cerebral thromboembolism had evidence of prothrombotic conditions. In particular, there was a predominance of children with anticardiolipin antibody (33%). These data support a recommendation that children with cerebral thromboembolism be evaluated for prothrombotic disorders. PMID- 9865799 TI - Cognitive loss in dementia with Lewy bodies and Alzheimer disease. AB - BACKGROUND: Dementia with Lewy bodies (DLB) is emerging as a common cause of degenerative dementia. Some preliminary evidence exists that the pattern of cognitive impairment in DLB is different from that in Alzheimer disease (AD). OBJECTIVE: To delineate features of cognitive impairment of DLB on standardized neuropsychological tests. METHODS: We performed neuropsychological assessments of 26 patients with probable DLB (based on criteria of the consortium on DLB international workshop) and of 52 patients with probable AD (based on criteria of the National Institute of Neurological and Communicative Disorders and Stroke [now the National Institute of Neurological Disorders and Stroke])-Alzheimer's Disease and Related Disorders Association) who were matched to the patients with DLB 2:1 by age, sex, education, and Mini-Mental State Examination score. RESULTS: Compared with the group with probable AD, the group with probable DLB scored significantly lower on the picture arrangement, block design, object assembly, and digit symbol substitution subtests of the Wechsler Adult Intelligence Scale Revised and on the Raven Colored Progressive Matrices test and significantly higher on the Mini-Mental State Examination locational orientation subtest and the Alzheimer's Disease Assessment Scale word recall subtest. A discriminant analysis revealed that the word recall score on the Alzheimer's Disease Assessment Scale and the block design score on the Wechsler Adult Intelligence Scale-Revised were the best discriminant factors. CONCLUSIONS: The disproportionately severe visuoperceptual, visuoconstructive, and visuospatial dysfunction and the disproportionately mild memory impairment in DLB compared with AD, which likely reflect the distribution of the pathologic changes in DLB, can help to differentiate DLB from AD. PMID- 9865800 TI - Is essential tremor symmetric? Observational data from a community-based study of essential tremor. AB - BACKGROUND: Essential tremor (ET) has been variably portrayed in the literature both as a symmetric arm tremor and as an asymmetric arm tremor. Few quantitative clinical or neurophysiological data specifically address the issue of tremor asymmetry in ET. OBJECTIVES: To examine a community-dwelling cohort of subjects with ET to (1) estimate the prevalence of tremor asymmetry and (2) quantify the magnitude of tremor asymmetry. METHODS: Fifty-four subjects with ET, identified in a community-based study of ET in New York City, underwent a Tremor Interview and a videotaped Tremor Examination. The examination included 6 tasks: sustained arm extension, pouring water, drinking water, using a spoon, finger-to-nose movements, and drawing spirals with each arm. Two neurologists rated the severity of tremor using a 0 to 3 clinical rating scale and a total tremor score was calculated (range, 0-36). Fourteen (25%) of 54 subjects also underwent quantitative computerized tremor analysis. RESULTS: The prevalence of asymmetry depended on the definition of asymmetry; small to moderate differences between sides were common. The mean side-to-side difference in clinical ratings for each of the 6 tasks was 0.54 of 3 points, which represented a 1.32-fold difference between sides. Clinical rating scores were higher in the nondominant arm in 39 subjects (72%), higher in the dominant arm in 9 (17%), and equal in 6 (11%). The 2 left-handed subjects had higher clinical ratings on the right. During quantitative computerized tremor analysis, there was a 1.71-fold mean difference between tremor amplitudes in the dominant and nondominant sides, and in 12 subjects (86%), the maximum tremor amplitude was in the nondominant arm. CONCLUSIONS: Small to moderate differences between sides were common in ET. In most community-dwelling subjects, tremor amplitude was greatest in the nondominant arm. In contrast, clinic-based studies have reported greater tremor in the dominant arm; those with ET who seek medical attention are more likely to exhibit severe tremor in their dominant arms. This study documents that mild asymmetry is a fundamental property of ET and that tremor is more severe in the nondominant arm. PMID- 9865801 TI - Evidence for cerebellar-frontal subsystem changes in children treated with intrathecal chemotherapy for leukemia: enhanced data analysis using an effect size model. AB - BACKGROUND: Following brain insult in early childhood, the later maturing neocerebellum and frontal lobes frequently show abnormalities. OBJECTIVE: To investigate the morphologic characteristics and function of a proposed cerebellar frontal subsystem in children treated for acute lymphoblastic leukemia (ALL) with intrathecal methotrexate using quantitative magnetic resonance imaging, neuropsychological measures, nonlinear multiple regression analysis, and a statistical effect size model that augments interpretive validity of nonsignificant statistical findings, particularly from small sample size studies. DESIGN: Comparison and relationship of magnetic resonance imaging morphometry of cerebellar lobuli I-V and VI-VII and prefrontal cortices, and performance on 5 neuropsychological tests assessing visual-spatial attention, short-term memory, and visuomotor organization and coordination between childhood survivors of ALL and a matched control group. PARTICIPANTS: Ten childhood survivors of ALL treated between 1982 and 1989 with standard 3-year intrathecal chemotherapy, and matched control subjects. MAIN OUTCOME MEASURES: Morphometric results of cerebellar lobuli I-V and VI-VII and prefrontal cortices, and results of Trail-Making Tests, Rey-Osterreith Complex Figure Test, WISC-III Coding. RESULTS: Significant effect size model values for outcome measures in the ALL group support deficits in lobuli VI-VII and prefrontal cortices, and neuropsychological performance. Multiple regression analysis results were consistent with hypothesized involvement of a cerebellar-frontal brain subsystem. CONCLUSION: Treatment of children with ALL with intrathecal methotrexate before 5 years of age has structural and functional effects on the developing neocerebellar-frontal subsystem. PMID- 9865802 TI - Familial cortical tremor, epilepsy, and mental retardation: a distinct clinical entity? AB - OBJECTIVE: To describe a European family with cortical tremor, epilepsy, and mental retardation, the pedigree of which indicates an autosomal dominant inheritance of the disease. DESIGN: Clinical, laboratory, neurophysiological, and neuroimaging data were studied. SETTING: Institute for research on mental retardation. PATIENTS: Two siblings (aged 25 and 28 years) and their 49-year-old mother had postural and action tremor, seizures, and mental retardation. Only tremor was present in the maternal grandmother (aged 68 years). The electroencephalogram showed diffuse spike-and-wave complexes and/or posterior spikes, and a photoparoxysmal response in the 4 subjects. The typical electrophysiologic features of cortical reflex myoclonus, such as giant somatosensory evoked potentials, enhancement of the C-reflex, and jerk-locked premyoclonus spikes, were found in all patients. CONCLUSION: This syndrome may represent a specific form of familial cortical tremor with a benign form of epilepsy and a new genetic model of cortical hyperexcitability inherited with an autosomal dominant mechanism. PMID- 9865803 TI - Partial status epilepticus associated with asomatognosia and alien hand-like behaviors [corrected]. AB - BACKGROUND: Paroxysmal alien hand syndrome (AHS) has been reported in association with ictal phenomena, but simultaneous electroencephalographic verification has not been made. OBSERVATION: A 61-year-old woman with a right hemisphere glioblastoma multiforme developed movements of the left hand that she claimed were not under her control, and she denied ownership of the affected limb. Simultaneous electroencephalograms documented continuous spikes in the right frontotemporal region. Intravenous diazepam therapy (2.5 mg) reversed both the abnormal movements and the spike activity on the electroencephalograms. CONCLUSIONS: Our case demonstrates that partial seizures can produce AHS along with asomatognosia. This variety of AHS appears to be different, both etiologically and phenomenologically, from other forms of AHS. PMID- 9865804 TI - Acute middle cerebral artery thrombosis demonstrated by cranial computed tomography: the "dense MCA" sign. PMID- 9865805 TI - Interferons in the treatment of multiple sclerosis: do they prevent the progression of the disease? PMID- 9865806 TI - Interferons should be used to treat most patients with MS. PMID- 9865807 TI - Interferons in the treatment of multiple sclerosis. PMID- 9865808 TI - 1860--neurology was there. AB - The year 1860 was a beginning of social, political, and medical upheaval in the United States. The Civil War over states' rights, which would last for 4 years and change the political landscape of the United States forever, had its roots in that year. American neurology was conceived and gestated by the Civil War because of the collaboration between William A. Hammond (1828-1900) and Silas Weir Mitchell (1829-1914), who became the founders of American neurology. Thus, 1860 serves as a pivotal point from which to view the precursors of American neurology and the later war and postwar developments. PMID- 9865809 TI - Limitations of acetaminophen, aspirin, and caffeine in alleviating migraine. PMID- 9865810 TI - Critical illness myopathy, steroids, and cytochrome P450. PMID- 9865811 TI - Depression and stimulant dependence: neurobiology and pharmacotherapy. AB - Depressive disorder rates in stimulant-dependent individuals are substantially higher than community rates. Further, depressive symptoms are considered a major component of stimulant withdrawal. The comorbidity of these disorders may reflect shared neurochemical alterations in the function of serotonin, dopamine, and peptide systems, such as corticotropin releasing factor (CRF) and neuropeptide Y (NPY). These alterations are observed in patients, and in animal models of depression and stimulant dependence, particularly in limbic brain structures. This shared neurobiology does not seem to result from significant shared heritability or genetic linkage; stimulants may induce changes in neurobiology that are similar to those found in depression, and these changes might provide a therapeutic target. Stimulant-dependent patients with a depressive disorder may be a specific subpopulation for antidepressant trials, and they might reduce their stimulant abuse when treated with antidepressants. Nevertheless, concomitant dependence on alcohol or opioids may influence this response, and antidepressants appear to be more effective for depression in combined stimulant and opioid dependence than in combined stimulant and alcohol dependence. PMID- 9865812 TI - P3a of event-related potential in chronic methamphetamine dependence. AB - To investigate the psychophysiological features of methamphetamine (MAP) dependence, we recorded auditory event-related potentials (ERPs) in 15 patients with MAP dependence and in 15 age-matched normal controls. ERPs were recorded during a standard oddball task and a read task similar to those employed by Squires et al. (Squires NK, Squires KC, Hillyard SA [1975] Two varieties of long latency positive waves evoked by unpredictable auditory stimuli in man. Electroenceph Clin Neurophysiol 38:387-401). The patients with MAP dependence showed reduced P3a amplitude and area in the read task and delayed P3b latency with normal P3b amplitude and area in the oddball task. These results suggest that central noradrenergic dysregulation may persist after the remission of acute psychotic symptoms in MAP psychosis and that chronic MAP dependence would produce impairment of the frontal cortex. PMID- 9865813 TI - Patient-treatment matching improves compliance of alcoholics in outpatient treatment. AB - In chronic medical disorders, patient compliance is very poor and often less than 50%. The purpose of this investigation was to determine whether the proper matching of patient, treatment, and therapist would improve patient compliance in the outpatient treatment of alcoholics. We randomly placed 119 alcoholics who had commenced protracted outpatient treatment for alcoholism in either low or high structured treatments. These 119 patients were offered 12 months of individual treatment. Interruption of this treatment was recorded as noncompliance. The treatment courses were expressed in survival curves. The consequence of compliance for long-term prognosis was described by means of the Addiction Severity Index. The form of treatment in itself had no importance for compliance. However, correct matching of a patient and a treatment structure resulted in a compliance rate of 63% as opposed to 38% compliance among mismatched patients. Twenty-four months after the initiation of treatment, patients in the compliance group had a significantly higher level of function than those in the noncompliance group. Our investigation of alcoholics involved in outpatient treatment shows that the correct matching of patients and treatment structure improves compliance and long-term prognosis. It is hoped that the present investigation will generate interest in future research on correct matching to gain improved compliance in patients with a chronic medical disorder. PMID- 9865814 TI - Communication disturbances and family psychiatric history in parents of schizophrenic patients. AB - Parents of schizophrenic patients have demonstrated subtle disturbances of thought, language, and communication. The etiologic relevance of these disturbances is not clear. This study assessed levels of referential communication disturbance in the "natural speech" of schizophrenic patients, nonschizophrenic parents of patients, and control subjects matched to the patients' parents and tested for associations of communication disturbances in parents with family history of psychosis and with schizotypy scale scores. The speech of the patients' parents as a group contained high frequencies of referential failures. Those parents with first-degree family histories of psychosis and/or high schizotypy scale scores made more frequent referential failures than the rest of the parents. Family history was particularly highly associated with failures involving language structural breakdown. The results of this study suggest that referential disturbances in parents of patients may be related to genetic liability in the parents. However, such an effect does not appear to account fully for the sizable differences between parents and controls in levels of communication disturbance. PMID- 9865815 TI - Gender differences in psychiatric illness and treatments: a critical review. AB - Controversy exists about the causation of gender differences long observed in the prevalence of mental disorders. Recent epidemiological, biochemical, and genetic research has shed further light upon both their etiologies and treatments. Both controversies and research are reviewed and critically examined. PMID- 9865816 TI - Are there cognitive subtypes in adult attention deficit/hyperactivity disorder? AB - There has been increasing knowledge of the treatment, diagnosis, and demographics of adults with residual attention deficit/hyperactivity disorder (ADHD). However, less is known about the neuropsychological functioning in adults with residual ADHD. In comparing the clinical neuropsychological test performance of a group of adult clinic patients with residual ADHD (N = 30) with that of normal controls (N = 10), we found the patients performed worse on the Trail Making Test, a visual continuous performance test, and the "Brown-Peterson" Auditory Consonant Trigrams Test, but not on any other neuropsychological measures. This pattern indicated a deficit in the area of executive control type functioning, a functional deficit that could be linked to dysregulation of frontal lobe brain systems. Of equal interest was that patients diagnosed with ADHD/hyperactive impulsive type (ADHD+) and patients diagnosed with ADHD/inattentive type (ADHD-) had different types of executive system deficits. ADHD+ was associated with relative deficiency on the Wisconsin Card Sorting Test. ADHD- was associated with relative deficiency on the "Brown-Peterson" Auditory Consonant Trigrams Test, a measure of working memory, as well as less olfactory identification on a smell identification test. The data are discussed in terms of recent localization theories of frontal lobe function. The preliminary data suggest that the different cognitive weaknesses of ADD subtypes may be linked to dysregulation of separate frontal brain regions and/or neurotransmitter systems. PMID- 9865817 TI - Diagnostic electron microscopy of neoplasms. PMID- 9865818 TI - Special techniques in diagnostic electron microscopy. AB - The power of electron microscopy as a diagnostic tool can be amplified considerably by the application of ancillary preparative and analytic methods. Subcellular chemistry and structure can be examined by various forms of microprobe analysis and by special staining methods, including cytochemical, immunocytochemical, and negative staining. Qualitative ultrastructural examination can be augmented by morphometric analysis. Correlative microscopic survey methods can be used as a means of targeting ultrastructural investigations. This article provides an overview of the use of these special techniques in the diagnosis and classification of tumors and other selected pathologic processes. PMID- 9865819 TI - Role of electron microscopy and other special techniques in the diagnosis of childhood round cell tumors. AB - A series of case presentations show unique challenges associated with childhood round cell tumors and the role of ancillary techniques in diagnosis. Electron microscopy is shown to be the most powerful individual technique. Immunohistochemistry is less effective but also essential. Other ancillary techniques may provide needed additional diagnostic information. Because this is an area where it is of great importance to secure the most rapid, accurate, and specific diagnosis possible, an integrated multimodal approach is recommended- incorporating light microscopic, electron microscopic, and immunohistochemical studies as a matter of routine, and providing for cytogenetic and/or molecular diagnostic studies as indicated. PMID- 9865820 TI - Cytology: screening or diagnostic tool? AB - Previously used only as a screening tool, cytology now emerges as a powerful diagnostic technique, especially since the advent of the fine needle aspiration (FNA) biopsy. This article highlights the use of ancillary techniques, primarily electron microscopy (EM), and immunohistochemistry (IHC). When coupled with routine cytological examination such as FNA and body cavity fluid cytology, EM and IHC can refine the diagnosis and make it more precise. The authors discuss how to solve common diagnostic dilemmas by the use of cytology along with IHC and EM. The following common diagnostic problems are addressed: mesothelioma versus adenocarcinoma, neuroendocrine neoplasms and their mimickers, melanoma versus carcinoma versus sarcoma, hepatocellular carcinoma versus adenocarcinoma and adenocarcinomas of unknown primary. PMID- 9865821 TI - Neuroendocrine carcinomas: role of immunocytochemistry and electron microscopy. AB - Neuroendocrine tumors occur in many sites of the body and can present significant diagnostic problems when poorly differentiated. To identify a tumor as neuroendocrine, pathologists commonly use either immunocytochemistry or electron microscopy. In this report, the various immunocytochemical reagents are reviewed along with the ultrastructural features of neuroendocrine tumors. Site-specific variations in neuroendocrine tumors are discussed. A cost-effectiveness evaluation was performed on tumors from one laboratory which showed that electron microscopy was a less expensive diagnostic modality if more than three antibodies were necessary to arrive at the correct pathological diagnosis. PMID- 9865822 TI - Role of electron microscopy in the evaluation of soft tissue neoplasms, with emphasis on spindle cell and pleomorphic tumors. AB - The current significant role of transmission electron microscopy in the evaluation of soft tissue tumors when correlated with conventional histological and immunohistochemical studies is discussed for the following entities: myxofibrosarcoma, storiform-pleomorphic fibrosarcoma (malignant fibrous histiocytoma), and myofibrosarcoma; dermatofibrosarcoma protuberans; hemangiopericytoma; monophasic synovial sarcoma; extrarenal rhabdoid tumor; soft tissue perineurioma; and gastrointestinal stromal tumors, notably the so-called autonomic nerve variant. PMID- 9865823 TI - Ultrastructural features of diffuse malignant mesotheliomas. AB - The distinction of malignant mesothelioma from tumors metastatic to the serosal membranes can often be made based on the results of histochemical or immunohistochemical studies. However, in some cases, these techniques are inadequate to make a firm diagnosis. In these instances, electron microscopic studies with the observation of a constellation of characteristic ultrastructural findings may permit an unequivocal diagnosis of mesothelioma. PMID- 9865824 TI - Metastatic adenocarcinoma of unknown primary origin. AB - Adenocarcinomas account for up to 60% of all metastatic neoplasms of unknown primary origin. In general, adenocarcinomas are the most difficult metastatic tumor to accurately identify the primary site. Some metastatic adenocarcinomas have distinctive histological features that allow for their site determination (eg, colonic adenocarcinoma, bronchioloalveolar cell carcinoma), although the majority of metastatic adenocarcinomas have histological features that are not distinctive enough to allow for a specific diagnosis of their origin. For this reason, electron microscopy and immunohistochemistry have been used to help identify the exact type (origin) of metastatic adenocarcinomas. Relatively specific ultrastructural features used to diagnose metastatic adenocarcinomas of unknown primary origin include tubular myelin, intranuclear surfactant apoprotein tubular inclusions, Clara cell granules, uniform short microvilli with filamentous cores and core rootlets, Langerhans cells associated with neoplastic cells, cytoplasmic hyaline globules, lipid droplets, glycogen, and cytoplasmic crystals. Only a few of these ultrastructural features are absolutely specific. Relatively specific immunohistochemical tests used to diagnose metastatic adenocarcinomas of unknown primary origin include prostate-specific antigen, thyroglobulin, estrogen and progesterone receptor proteins, thyroid transcription factor-I, and surfactant apoproteins. Of these, prostate-specific antigen and thyroglobulin are the most specific. The purpose of this article is to discuss the use of electron microscopy and immunohistochemistry in the site-specific diagnosis of metastatic adenocarcinomas of unknown primary origin. PMID- 9865826 TI - Clinicopathologic analysis of microscopically invasive breast carcinoma. AB - Breast biopsy or mastectomy cases having diagnoses of carcinoma in situ with "microinvasion," "minimal invasion," "focal invasion," or "suggestive of invasion" were reviewed and all histologically identified foci of invasive disease from each case were measured using an ocular micrometer. Cases in which any single focus of invasion was greater than 5 mm or the added size of separate invasive foci exceeded 10 mm were excluded, resulting in a study group of 75 patients. Invasive neoplasm was present in the initial biopsy in 69 of 75 cases (92%); however, residual invasive neoplasm was found in the subsequent lumpectomy/mastectomy from 14 of these (20%). In 59% of cases, two or more histologically separate foci of invasion were identified. Invasive foci consisted of isolated cells or cell clusters, each less than 1 mm (microfocal invasion), in 33% of cases. In 12 cases, the sum of individual invasive foci was 5 to 10 mm. Axillary lymph nodes (LN) from 5 of 69 patients (7%) contained metastatic carcinoma (four cases, one LN positive; one case, two LN positive). The cumulative sizes of all invasive foci in the LN-positive group were microfocal invasion (one case), 0.6 mm (one case), 1.1 mm, 2.5 mm, and 5.8 mm. The difference in frequency of axillary node metastasis between tumors with microfocal and measurable invasion (4.3% v 8.6%) was not statistically significant. Follow-up data were available on 55 cases (mean interval, 66.1 months). One (node-negative) patient had duct carcinoma in situ recurrence in the same breast 4 years after initial treatment. Another (with unknown node status) developed an axillary lymph node metastasis 13 months after initial treatment (96% disease-free survival). We conclude that microscopic stromal invasion in breast carcinoma, at least in the setting of significant in situ component, is often initiated from multiple foci. Patients with microscopically invasive breast carcinoma have a small but significant risk of axillary metastases, although a highly favorable survival. PMID- 9865827 TI - MIB-1 labeling indices in benign, aggressive, and malignant meningiomas: a study of 90 tumors. AB - Predicting tumor behavior in meningiomas based on histology alone has been problematic. This study retrospectively compares histology and MIB-1 (cell proliferation marker) labeling indices (LI) in benign, aggressive, and malignant meningiomas. Six histological features, including mitoses, necrosis, loss of pattern, hypervascularity/hemosiderin deposition, prominent nucleoli, and nuclear pleomorphism, were compared in 90 meningiomas (Fisher's exact test). Tumors with two or more of the above features were designated as aggressive meningiomas. Malignant meningiomas were characterized by brain invasion or metastasis. The MIB 1 LIs (% positive tumor cell nuclei) were compared between the three groups (Kruskal-Wallis test, Wilcoxon two-sample test). Of the benign meningiomas (n=37; mean age, 54 years), 41% had one of the six histological features, with nuclear pleomorphism (n=10) being the most frequent. The aggressive tumors (n=29; mean age, 61 years) were characterized by nuclear pleomorphism (n=28), mitoses (n=20), necrosis (n=16), loss of pattern (n=16), prominent nucleoli (n=6), and hypervascularity/hemosiderin deposition (n=5). Malignant tumors (n=24; mean age, 59 years) were characterized by nuclear pleomorphism (n=22), mitoses (n=21), loss of pattern (n=21), necrosis (n=21), nucleoli (n=17), and hypervascularity/hemosiderin deposition (n=3). Significant differences were found between the aggressive and malignant groups with regard to loss of pattern, necrosis, and nucleoli (P=.0043, .011, and .00029, respectively). Mean MIB-1 LIs for the benign, aggressive, and malignant groups were 1.0% (range, 0 to 5.5%),5.5% (range, 0.1 to 32.5%), and 12.0% (range, 0.3 to 32.5%), respectively. Differences in the mean MIB-1 LI between groups were statistically significant, with P values of <.0001 (benign v aggressive) and .0012 (aggressive v malignant). Mean MIB-1 LIs for recurrent versus nonrecurrent tumors were 7.1% (range, 0 to 32.5%) versus 3.8% (range, 0 to 20.9%) (P=.32). The mean MIB-1 LI for patients who were alive with or without tumor was 6.2% (range, 0 to 32.5%) versus a mean MIB-1 LI of 14.2% (range, 2.8% to 32.5%) for patients who died of or with tumor (P=.0013). In conclusion, (1) There is a statistically significant difference in the increasing MIB-1 LI means between benign, aggressive, and malignant meningiomas and between patients who were alive versus those who died; (2) there is some overlap in MIB-1 LI ranges between groups, which warrants caution in interpreting an individual MIB-1 LI in a given tumor. PMID- 9865825 TI - Electron microscopy in tumor diagnosis: indications for its use in the immunohistochemical era. AB - The electron microscope reveals much more information on tumor cell structure than can be obtained with the light microscope, and some of the data are useful in diagnostic studies. In this review, major contributions of electron microscopy in the main tumor categories are discussed. To select between immunocytochemistry and electron microscopy, the probable contributions of each in the context of the particular case must be assessed. Usually, electron microscopy will only be requested after a battery of immunostaining procedures has been performed and found to be insufficient, but there are occasions when ulstrastructural study is logically the first choice after routine light microscopy. It is worth taking tissue for possible electron microscopy from any tumor that is known or anticipated to be a diagnostic problem. PMID- 9865828 TI - Sex and androgenic steroid receptor expression in hepatic adenomas. AB - Sex hormones and anabolic-androgenic steroids are implicated in the development and progression of hepatic adenomas (HA). We studied the expression of their receptors in HA and adjacent liver. Archival tissue sections of 27 HA (16 resections, four needle biopsies, seven aspirations) from 18 patients, and the adjacent liver, were immunostained with monoclonal antibody to estrogen receptor (ER, 1/80) (Dako, Carpinteria, CA), progesterone receptor (PR, 1/50) (BioGenex, San Ramon, CA), and androgen receptor (AR, 1/80) (BioGenex). An avidin-biotin complex technique was used with microwave antigen retrieval. Nuclear expression was assessed as 1+ to 3+ intensity, with semiquantitation of the percentage of nuclei immunopositive. Five percent or more nuclei immunopositive was regarded as positive. The 18 patients included 16 females of 34 years mean age (range, 16 to 49) with an available history of oral contraceptives in five; the two men were 24 and 30 years, with no history of androgenic steroids. ER, PR, and AR were present in seven (26%) (1+/-2+ intensity, 5% to 10% of nuclei) of HA, seven (26%) (1+/-2+ intensity, 5% to 30% of nuclei) and nine (33%) (1+/-3+ intensity, 5% to 80% of nuclei), respectively. In the adjacent liver in 11 cases, there were one (9%) ER, (2+ intensity, 5% of nuclei), four (36%) PR (1+/-2+ intensity, 5% to 20% of nuclei), and two (18%) AR (2+/-3+ intensity, 10% of nuclei). Receptors are present and may mediate the action of sex hormones or androgenic steroids on HA and adjacent liver, but in less than one third of patients. This may have therapeutic implications. PMID- 9865829 TI - Colonic histopathology in untreated celiac sprue or refractory sprue: is it lymphocytic colitis or colonic lymphocytosis? AB - Colonic histopathology in some patients with untreated celiac sprue and refractory sprue has been said to be indistinguishable from lymphocytic colitis, but there have been no objective comparisons on which this is based. The purpose of this study was to determine the prevalence and to characterize the nature of colonic histopathology at the time of diagnosis in patients with celiac or refractory sprue. Colonoscopic biopsy specimens obtained at the time of diagnosis from 16 patients with celiac sprue, six patients with refractory sprue, nine patients with lymphocytic colitis, and five normal controls were analyzed blindly by histological and morphometric methods, quantitating the number and specific subtypes of inflammatory cells within the lamina propria and epithelium. Immunoperoxidase staining of intraepithelial lymphocytes with a monoclonal antibody to CD8 also was performed. Three of 16 patients with untreated celiac sprue (19%) were thought to have colonic histological abnormalities, which by morphometry consisted of slightly increased numbers of lymphocytes in the surface epithelium and lamina propria, many of which were CD8-positive. These abnormalities were distinguishable from lymphocytic colitis by the lack of increased overall lamina propria cellularity and surface epithelial abnormalities, and by fewer intraepithelial lymphocytes. In refractory sprue, colonic histological abnormalities were more frequent than in celiac sprue, occurring in four of six patients (67%), more pronounced, and identical to those in the lymphocytic colitis syndrome. However, colonic intraepithelial lymphocytes in lymphocytic colitis were mostly CD8-positive, whereas those in the colitis of refractory sprue rarely were. Mild colonic lymphocytosis in patients with untreated celiac sprue should be distinguished from lymphocytic colitis by the lack of surface epithelial abnormalities, the lack of increased cellularity of the lamina propria, and the lack of ongoing watery diarrhea after treatment with a gluten-free diet. In contrast, colonic histopathology in refractory sprue is indistinguishable from lymphocytic colitis, although immunohistochemical differences do exist. PMID- 9865830 TI - Immunoreactivity for LN2 and LN3 distinguishes small cell carcinomas from non small cell carcinomas in the lung. AB - Immunoreactivity to LN2 and LN3, monoclonal antibodies that recognize components of the class II major histocompatibility complex, was assessed in 72 cases of non small cell lung carcinoma (NSCLC) (32 biopsy specimens, 40 resection specimens) and 64 cases of small cell carcinoma (56 biopsy specimens, 8 resections) of the lung. All cases were reviewed independently by three pathologists for histological classification. Only 1 of the 64 small cell carcinomas showed immunoreactivity for LN2, and none of the 64 cases showed reactivity for LN3. Among the non-small cell carcinomas, 25 of 48 cases were positive for LN2 and 43 of 71 were positive for LN3; the sensitivity was greater for adenocarcinoma (78.5%) than for squamous cell carcinoma (37%). A combined sensitivity of 64.7% was observed when the results of LN2 and LN3 were combined, and this sensitivity was not significantly diminished in the biopsy subset of cases (59.4%). Differentiation within histological subtypes of NSCLC (ie, well, moderate, or poorly differentiated) did not alter test sensitivity. In conclusion, LN2 and LN3, used alone or in combination, appear highly specific for non-small cell carcinoma and moderately sensitive in both biopsy and resection specimens; therefore, these antibodies may be diagnostically useful in distinguishing small cell from non-small cell carcinoma of the lung. PMID- 9865831 TI - Distinction of basaloid carcinoma of the prostate from benign basal cell lesions by using immunohistochemistry for bcl-2 and Ki-67. AB - The distinction of rare basaloid carcinomas (BC) of the prostate from more common basal cell hyperplasia may be difficult, because basal cell hyperplasia (BCH) may have prominent nucleoli and may appear infiltrative. Using immunohistochemistry, we studied bcl-2 and p53 expression and Ki-67 proliferation index in eight cases of typical BCH, eight cases of BCH with nucleoli, and six cases of BC. Bcl-2 expression (P < .0001) and Ki-67 index (P=.005) were elevated in BC compared with typical BCH or BCH with nucleoli, whereas there was no significant difference between typical BCH and BCH with nucleoli. P53 was not discriminative in separating benign from malignant basal cell lesions of the prostate. Bcl-2 may play a role in the pathogenesis of basal cell lesions of the prostate. Elevated expression of bcl-2 and higher Ki-67 index may aid in the diagnosis of basal cell proliferative lesions of the prostate. PMID- 9865832 TI - Clear cell adenocarcinoma and nephrogenic adenoma of the urethra and urinary bladder: a histopathologic and immunohistochemical comparison. AB - Because of histological similarities between nephrogenic adenomas and clear cell adenocarcinomas of the urinary tract, there is the potential for diagnostic confusion between these two entities. The histopathologic features of 13 nephrogenic adenomas and five clear cell adenocarcinomas of the urethra and urinary bladder are compared in this report, and detailed immunohistochemical staining profiles are provided for these tumors. Only 2 of the 13 nephrogenic adenomas contained clear cells, and these constituted less than 10% of the lesions. In contrast, four of the five clear cell adenocarcinomas contained prominent areas with clear cells. Nephrogenic adenomas generally showed only mild cytologic atypia, whereas four of the five clear cell adenocarcinomas showed severe atypia. A single mitotic figure was identified in only two of the nephrogenic adenomas, whereas the mitotic rate in the clear cell adenocarcinomas ranged from 2 to 14 per 10 high-power fields. None of the nephrogenic adenomas showed evidence of necrosis, but focal necrosis was noted in four of the five clear cell adenocarcinomas. In general, the nephrogenic adenomas and clear cell adenocarcinomas showed negative to weak staining with CK903 but strong staining with AE1, AE3, and Cam 5.2. Variable staining was observed with Brst-3 and antibodies to S-100, CEA (monoclonal and polyclonal), LeuM-1, and CA19.9. Nephrogenic adenomas and clear cell adenocarcinomas were all negative for prostate-specific acid phosphatase (PSAP), prostate-specific antigen (PSA), and estrogen and progesterone receptors (except for two nephrogenic adenomas, which showed only focal weak staining for estrogen receptor). Neither bcl-2 nor c-erbB 2 staining was able to discriminate between the tumors. However, strong staining for p53 was noted in each clear cell adenocarcinoma and in none of the nephrogenic adenomas. MIB-1 positivity in nephrogenic adenomas ranged from 0 to 13 (average of 5.5) per 200 cells, whereas the positive range for clear cell adenocarcinomas was 33 to 70 (average of 47) per 200 cells. In summary, histopathologic features that favor clear cell adenocarcinoma over nephrogenic adenoma include a predominance of clear cells, severe cytological atypia, high mitotic rate, necrosis, high MIB-1 positivity, and strong staining for p53. PMID- 9865833 TI - Chronic chorioamnionitis: a clinicopathologic and immunohistochemical study. AB - Lymphocytic inflammation of the fetal membranes is unusual and has been termed chronic chorioamnionitis. We report the clinicopathologic and immunohistochemical findings in 31 placentas with chronic chorioamnionitis. The most common histopathologic association was chronic villitis of unknown etiology, which was identified in 22 (71%) of the 31 placentas. The severity of the chronic villitis did not correlate with the severity of chronic chorioamnionitis. Additional placental findings included chronic intervillositis in two, fetal vessel thrombosis in five, hemorrhagic endovasculitis in four, decidual chronic vasculitis in three, and atherosis in one. Maternal history included pregnancy induced hypertension in six and diabetes in one. Twelve infants were preterm, and five had intrauterine growth retardation. There was no neonatal sepsis or death. Immunohistochemical staining in areas of chronic chorioamnionitis showed CD3+ and CD8+ cells present in moderate numbers, and CD4+ cells in smaller numbers. CD20+ and CD56+ cells were rare or absent. Chronic chorioamnionitis is commonly associated with chronic villitis of unknown etiology, shares similar clinical associations, and may have a related cause, possibly immunologic. PMID- 9865834 TI - Morphometric grading in breast cancer: thresholds for mitotic counts. AB - Three hundred sixty-four cases of invasive ductal breast cancer diagnosed during the years 1988 to 1991 were analyzed to determine quantitative thresholds for mitotic activity. Mitotic counts were calculated in each sample and expressed as standardized mitotic index (SMI) and mitotic activity index (MAI). Based on Kaplan-Meier curves, univariate and multivariate analysis of Cox's regression, and maximum efficiencies of ROC analysis, optimal thresholds were determined on the basis of survival and recurrence of disease. In our material, with a follow up time of 5 years 9 months, we found two thresholds--a lower and a higher--for both SMI (17 mitoses/mm2 and 32 mitoses/mm2) and MAI (13 mitoses/10 HPF and 35 mitoses/10 HPF). The thresholds were the same in the whole material and in subgroups divided according to the patients' age and axillary lymph node status at the time of diagnosis, and tumor size. The thresholds clearly separated patients with favorable, intermediate, and unfavourable outcome of disease. In our material, the risk of breast cancer death associated with the determined thresholds (ranging from 4.7 to 3.8) clearly exceeded those of menopausal status, axillary lymph node status and tumor size. The risk of breast cancer death associated with the determined thresholds was still emphasized in the groups of premenopausal and axillary lymph node-negative patients, and with tumor size less than 2 cm in diameter (risk ratios, 11.8, 6.0, and 6.7, respectively). The results suggest that the presented quantitative thresholds could be applied in grading of invasive ductal breast cancer. PMID- 9865835 TI - The necessary role of the autopsy in cardiovascular epidemiology. AB - The autopsy rate in the United States today is remarkably low, with proportionally fewer autopsies for natural causes of death. Consequently, most cardiovascular epidemiology studies do not use autopsy data and rely on death certificates, medical records, questionnaires, and family interviews as sources of mortality information. These practices introduce a high degree of variability and uncertainty regarding cause of death. This review illustrates the necessity for increased use of autopsies in cardiovascular epidemiology by critically evaluating other measures of cardiovascular disease (CVD) incidence. We evaluated the literature regarding CVD as cause of death and conducted discussions with cardiologists, pathologists, and epidemiologists. No attempt was made for meta analysis. This review shows the limited reliability of death certificates, medical records, and interviews as sources of mortality statistics. In addition, the autopsy's role in clearly indicating the presence of CVD is illustrated. The autopsy used in conjunction with medical records is the only reliable means for establishing cause of death from CVD. There is an urgent need to reassess the current dependence of statistical mortality data on death certificates and other inadequate sources of CVD incidence. Death certificates, in general, are inadequately monitored for quality control and appropriate administrative oversight. With an increase in the number of hospitals performing no autopsies to investigate cause of death, a uniform national autopsy database is needed. PMID- 9865836 TI - Enhanced expression of Ki-67, topoisomerase IIalpha, PCNA, p53 and p21WAF1/Cip1 reflecting proliferation and repair activity in UV-irradiated melanocytic nevi. AB - To investigate the effect of ultraviolet (UV) irradiation on the expression of cell cycle-associated proteins, melanocytic nevi from healthy volunteers were partially covered, irradiated with a defined UV dose, and excised 1 week thereafter. The irradiated and the protected parts were examined separately by conventional microscopy and immunohistochemistry using the antibodies Ki-S11 (Ki 67), Ki-S7 (topoisomerase IIalpha), PC10 (proliferating cell nuclear antigen [PCNA]), DO-7 (p53), 6B6 (p21WAF1/Cip1), and the melanocytic marker HMB-45. DNA nick-end labeling was used as a marker of apoptosis. Irradiation resulted in morphological changes and increased HMB-45 reactivity. Proliferation, as assessed by Ki-67 and topoisomerase IIalpha expression, was also clearly enhanced in the UV-exposed areas. This was confirmed by the appearance of occasional mitotic figures. PCNA expression levels markedly exceeded those of the proliferation markers and did not correlate with the latter in most cases. p21 immunolabeling indices were also consistently augmented after UV exposure; hence it is likely that growth-inhibitory mechanisms partly compensate for the proliferative impulse, and the disproportional rise in PCNA expression probably reflects DNA repair activity. Enhanced p53 immunostaining in four cases suggests that the induction of p21 after irradiation may be p53 mediated, whereas no concomitant apoptotic events were observed. We conclude that UV light can stimulate the proliferative activity of melanocytes in melanocytic nevi, but that simultaneously cell cycle inhibitors are activated to permit DNA repair. PMID- 9865837 TI - Characterization of cytoplasmic secretory granules (PSG), in prostatic epithelium and their transformation-induced loss in dysplasia and adenocarcinoma. AB - Cytoplasmic clarity is a histological feature of normal prostatic secretory cells, but in this study, tissue fixation in strong (>2.5%) glutaraldehyde dramatically altered cytological staining. Secretory cytoplasm appeared red and granular on routine stains because of myriad intensely staining eosinophilic granules (PSG). Immunostaining for prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) showed their exclusive localization to the PSG. Electron microscopy confirmed these findings and also showed that after fixation in many agents, including formaldehyde, PSG appeared empty, accounting for the artefactual "clear cell" appearance on light microscopy. PSG were most densely concentrated apically in a bud-shaped luminal compartment in which cytokeratin was selectively absent. Normal exocrine secretion was visualized as detachment of apocrine buds or their in situ disintegration. Distinctively in dysplasia and almost all carcinomas, PSG were rare to absent, and proteases were free in the cytoplasm, often concentrated beneath the apical membrane. The apocrine compartment was absent, with no observed secretory mechanism. Tumor cells had dark amphiphilic cytoplasm after all fixatives. This provided a reliable method of distinguishing malignant from benign glands in tissues fixed in strong glutaraldehyde. Clear cell carcinomas, whose cytoplasm mimicked routinely fixed normal secretory cells, surprisingly had almost no PSG. Instead, their "granules" were lipid-filled vacuoles reflecting a secretory pathway not seen in normal cells, dysplasia, or the common "dark cell" carcinomas. These observations may define two distinctive biological pathways of prostate cancer evolution and may facilitate diagnostic decisions on needle biopsy samples. PMID- 9865838 TI - Pleuro-pulmonary endometriosis and pulmonary ectopic deciduosis: a clinicopathologic and immunohistochemical study of 10 cases with emphasis on diagnostic pitfalls. AB - The clinical and pathological features of nine cases of pleuro-pulmonary endometriosis and the first case of pulmonary ectopic deciduosis are presented. The patients were all women between the ages of 27 and 74 years (median, 36 years) who presented with symptoms of catamenial pleural pain, shortness of breath, hemoptysis, or radiographically detected lung masses. Clinically, six patients were multiparous, one patient had pelvic endometriosis, and four patients had undergone prior pelvic surgical procedures, including dilatation and curettage. Radiographically, eight patients had pulmonary infiltrates or nodules, and four patients had pneumothorax. Three cases involved the visceral pleura and one case the parietal pleura. The other six cases, including the single case of ectopic deciduosis, involved the lung parenchyma. Histologically, the single or multifocal lesions were well circumscribed or infiltrative, nodular, cystic, or nodulo-cystic, and showed the characteristic features of proliferative or secretory endometrium with numerous mullerian metaplastic changes. Mucin stains were negative in five cases of endometriosis and in the single case of ectopic deciduosis. Immunohistochemical studies were performed in these same six cases using antibodies to epithelial, mesenchymal, vascular, and neuroendocrine markers. The glandular epithelium was decorated with antibodies to pan cytokeratin, CK7, BER-EP4, ER, and PR, whereas the stromal cells showed positive staining for vimentin, actin, smooth muscle actin, desmin, ER, and PR. Follow-up information obtained in seven patients showed all women without recurrences after 1 to 20 years. The current study highlights the importance of recognizing intrathoracic endometriosis and ectopic deciduosis and properly assessing small biopsy specimens to avoid a misdiagnosis of malignancy. PMID- 9865839 TI - Calcifying aponeurotic fibroma: a clinicopathologic study of 22 cases arising in uncommon sites. AB - Calcifying aponeurotic fibroma is a rare soft tissue tumor that primarily occurs in children and adolescents and has a strong predilection for the distal portion of the extremities, especially the hands and feet. This report describes 22 previously unpublished cases arising in uncommon sites. Fifteen patients were male, and seven were female (age range, 2 to 43 years; median age, 9 years). The process typically presented as a painless mass and was present from 2 weeks to 11 years before resection. Sites of involvement were the back (n=8), knee region (n=5), thigh (n=3), forearm (n=3), elbow (n=2), and arm, not otherwise specified (n=1). The lesions were often adherent to dense fibrous connective tissue (eg, tendon, fascia, or periosteum) and ranged from 1.0 to 5.0 cm in maximum dimension. The process typically had an irregular contour and a firm, fibrous consistency. Sometimes minute foci with a calcific appearance were evident grossly. Microscopic examination showed spindled fibroblasts with a fascicular growth pattern and scattered epithelioid cells bordering chondroid foci with or without mineralization. Immunoreactivity was present for vimentin (six of six), muscle-specific actin (three of six), smooth muscle actin (three of six), CD99 (five of five), CD34 (one of six), CD57 (one of six, trace), EMA (two of six, trace), S100 protein (five of six), CD68 (five of five), and progesterone receptor (one of six). The tumors were managed by local excision (n=20), incomplete local excision (n=1) and biopsy only (n=1). Follow-up information was available for 10 patients with a median follow-up interval of 94 months. Five patients (50%) developed one or more recurrences. Familiarity with this entity should help to avoid confusion with other processes, including infantile and extraabdominal fibromatoses, a chondroma of soft parts, and a fibrous hamartoma of infancy. PMID- 9865840 TI - Cyclin D1 and MIB-1 immunohistochemistry in pilocytic astrocytomas: a study of 48 cases. AB - Pilocytic astrocytoma is an infrequently encountered, generally low-grade neoplasm. No study has extensively looked at both cyclin D1 and MIB-1 labeling indices in pilocytic astrocytoma and their relation to clinical outcome. This study retrospectively examines the clinicopathologic features of 48 patients with pilocytic astrocytoma including MIB-1 (cell proliferation marker) and cyclin D1 (protein that regulates progression from G1 to S phase of the cell cycle) immunohistochemistry. Of 48 patients (27 females and 21 males; mean age, 12.7 years; age range, 2 to 57 years), 26 initially underwent gross total resection; 17, subtotal resection; four, biopsy alone; in one patient, the extent of tumor resection was unknown. Histological features observed included Rosenthal fibers (83.3%), granular bodies (75%), vascular sclerosis (56.2%), vascular proliferation (56.2%), prominent nuclear pleomorphism (14.6%), necrosis (10.4%), and identifiable mitotic figures (2.1%). MIB-1 labeling indices (n=45) (positive staining tumor nuclei per 1,000 nuclei evaluated) ranged from 0 to 3.5% (mean, 0.6%); seven tumors had a labeling index greater than 1.0%. Cyclin D1 labeling indices (n=45) ranged from 0 to 0.8% (mean, 0.1%). Most tumors (N=29, 66.7%) had no immunostaining. At last known follow-up, 27 patients were alive with no evidence of disease (mean, 49.2 months), 17 patients were alive with evidence of disease (mean, 36.8 months), three died with tumor at 2, 22, and 156 months, and one patient was lost to follow-up. Eight patients had at least one tumor recurrence requiring additional surgery; seven of these patients had an initial subtotal resection. In summary, MIB-1 labeling indices were generally low (mean, 0.6%) and are reflective of the slow growth of the tumors. Cyclin D1 immunostaining does not appear to be significantly increased in pilocytic astrocytoma. Adverse outcome in patients with pilocytic astrocytoma may be related to extent of surgical resection and does not seem to correlate with histology, MIB-1 labeling indices, or cyclin D1 immunoreactivity. PMID- 9865841 TI - Altered mucin expression is a field change that accompanies mucinous (colloid) breast carcinoma histogenesis. AB - Mucinous carcinomas of the breast, so-called colloid carcinomas, exhibit better prognoses than their nonmucinous breast counterparts. This biological difference exhibited by mucinous breast carcinomas prompted us to examine the relationship of mucin expression to colloid carcinoma histogenesis. We studied 50 colloid carcinomas, 50 noncolloid cancers, and 50 normal breasts by hematoxylin-eosin (H&E) and Alcian blue staining, mucin immunohistochemistry, in situ hybridization with a battery of MUC riboprobes, and ancillary digital image analysis. We observed luminal mucin in normal ducts in 80% of colloid carcinomas compared with 10% of noncolloid carcinomas and 6% of normal breasts (P < .01). In the cases of colloid carcinoma that showed mucin-filled ducts, luminal mucin was observed in 40% of the normal ducts and acini, 40% to 75% of the ducts involved by hyperplasia, atypical ductal hyperplasia (ADH) and ductal carcinoma in situ (DCIS), respectively, and in 50% of the co-incidental areas of cysts (mucoceles), adenosis, fibroadenoma, and intraductal papilloma (P < .01). Immunohistochemistry showed that colloid carcinomas showed strong MUC2 cytoplasmic immunoreactivity and decreased MUC1 immunoreactivity compared with noncolloid carcinomas. In situ hybridization studies indicated fivefold increased MUC2 signals and twofold increased MUC5 signals within adjacent and remote normal epithelium in only the colloid carcinoma cases (P < .01; P < .05). In these cases of colloid carcinoma, these increased MUC2 and MUC5 signals were also observed in areas of hyperplasia, ADH, DCIS, and invasive carcinoma. In contrast, the noncolloid carcinomas showed fivefold increased MUC1 signals but no increases in MUC2 or MUC5. In mixed colloid/noncolloid carcinomas, the colloid areas had identical mucin expression patterns as the pure colloid carcinomas, but there was a loss of MUC2 and MUC5 expression and a gain of MUC1 expression in the noncolloid areas that was therefore identical to the pattern observed in pure noncolloid carcinoma. In this study, we conclude that the altered expression of mucin so characteristic of colloid carcinoma is also a field change present in adjacent and remote normal breast epithelium. PMID- 9865842 TI - Maternal scleroderma: placental findings and perinatal outcome. AB - Pregnancy after the onset of scleroderma is uncommon; therefore, placental findings and perinatal outcome have rarely been correlated. The histopathologic features of placentas from 13 pregnancies in eight women with scleroderma were recorded and correlated with the clinical features of the mother and fetus. Adverse perinatal outcome included intrauterine fetal demise in five, and previable or preterm delivery in four. A decidual vasculopathy was seen in 5 of the 13 placentas, four of which were associated with intrauterine fetal demise. Decidual blood vessels in the scleroderma patients were evaluated immunohistochemically for platelet-derived growth factor (PDGF), transforming growth factor beta1 (TGF-beta1), T-helper and T-suppressor lymphocytes, macrophages, immunoglobulin (Ig) M, and IgG, and compared with those from hypertensive and uncomplicated third-trimester pregnancies. The atherotic blood vessels in scleroderma were characterized by mural macrophages and IgM and IgG deposition and were similar to those seen in placentas from hypertensive pregnancies. CD8-positive T cells predominated in normal and hypertensive decidua compared with scleroderma, in which CD4-positive T cells were more frequent. No difference in PDGF or TGF-beta1 staining was found between scleroderma and control groups. In conclusion, decidual vasculopathy is common in scleroderma, is similar to that seen in hypertension, and is associated with poor perinatal outcome. A trend toward a reversed ratio of decidual CD4 to CD8-positive T cells is seen in scleroderma compared with hypertension and uncomplicated pregnancies. PDGF and TGF-beta1 do not appear to be involved in the pathogenesis of decidual vasculopathy in scleroderma. PMID- 9865843 TI - Increased stem cell somatic mutation in the non-neoplastic colorectal mucosa of patients with familial adenomatous polyposis. AB - Colorectal tumorigenesis in familial adenomatous polyposis (FAP) results from somatic mutation of either the normal APC allele or another growth control gene in epithelial cells bearing a germline APC defect. The rate at which tumors develop is therefore dependent on the somatic mutation frequency; it is not known whether this is normal or elevated in FAP. We aimed to quantify stem cell somatic mutation in FAP, comparing it with hereditary nonpolyposis colorectal cancer (HNPCC) and Crohn's disease (CD). Stem cell somatic mutation frequency was studied in 47 FAP patients, 5 HNPCC patients, and 13 CD patients, all younger than 49 years, by quantifying crypt-restricted loss of O-acetyltransferase activity in sections of morphologically normal colonic mucosa from individuals heterozygous for this monogenically inherited polymorphism. Median stem cell somatic mutation frequency was significantly higher in FAP than HNPCC (4.2 x 10( 4) v 1.4 x 10(-4), Mann-Whitney U, P < .02). The level in CD (4.0 x 10(-4)) was similar to FAP. Mutated crypts occurred in groups more frequently in FAP (22%) than HNPCC (12%) or CD (10%), suggesting an increase in stem cell division associated with crypt fission in FAP. We conclude that stem cell somatic mutation frequency is raised in non-neoplastic colorectal mucosa in FAP. This is probably related to increased stem cell proliferation and contributes to the high rate of tumor formation in this condition. PMID- 9865844 TI - The association of selected pathological features with prostate cancer in a single-needle biopsy accession. AB - Isolated high-grade prostatic intraepithelial neoplasia (PIN) has been shown to be a positive predictor of prostate cancer (PCa) on follow-up biopsy. However, the incidence of isolated high-grade PIN in needle biopsy specimens has been reported with a highly variable frequency of 1% to 15%. The current study examined the relationship of various pathological features with PCa on a single biopsy accession. A study population of 388 community-based consecutive needle biopsy accessions was prospectively recorded by a single pathologist (T.M.W.). All of the individual biopsy specimens were coded for the presence of PCa, high grade PIN, low-grade PIN, chronic inflammation (CI), intraluminal prostatic crystalloids (IPC) in benign glands, and mucinous metaplasia (MM). One hundred twenty-nine (33%) of the patients were diagnosed with PCa. The 8% incidence of isolated high-grade PIN was consistent with previous studies. The incidence of other pathological features were as follows: high-grade PIN, 14%; low-grade PIN, 13%; CI, 30%; IPC, 4%; and MM, 8%. Of the patients with high-grade PIN, 47% had PCa on a separate core biopsy, whereas 31% of patients without high-grade PIN were observed to have PCa (P=.021). Of the patients with CI, 21% were found to have PCa on a separate core, whereas 38% of patients without CI were found to have PCa (P=.0009). None of the other pathological features surveyed showed any significant association with PCa. High-grade PIN was a relatively common finding (14%) in this study and was positively associated with PCa on a separate core from the same accession biopsy. The negative association of CP with PCa within the same accession has not been reported previously and may be an artifact related to the clinical indications for a prostatic biopsy. PMID- 9865845 TI - Expression of proliferation and apoptosis-related proteins in usual ductal hyperplasia of the breast. AB - Expression of proliferation- and apoptosis-related proteins was studied by immunohistochemistry in 130 usual ductal hyperplasias of the breast, of which 39 cases (30%) had adjacent invasive cancer. Overexpression of cyclin D1 and Ki-67 was found in 6% and 29% of the cases, respectively. Only two mild ductal hyperplasias were Her-2/neu positive. Overexpression of p21 and reduced expression of p27, both cdk-inhibitors, was seen in 16% and 27% of the lesions, respectively. Reduced expression of bcl-2 was found in 16% of the cases, and p53 accumulation was present in 8%. Expression of six of the seven studied proteins showed no significant difference between mild, moderate, or florid ductal hyperplasias, indicating that there are no important cell biological differences with regard to the studied proteins between the lesions within this morphologically continuous spectrum. In addition, there were no differences between lesions with and without an invasive component. Cyclin D1 positivity was exclusively seen in lesions with 75% or more p27-positive nuclei. No significant correlations were found between other proteins. Twenty-three of 91 lesions (25%) had multiple events, of which five showed altered expressions of three or four proteins. In conclusion, altered protein expression of several proliferation- and apoptosis-related genes that are known to be involved in invasive breast cancer also may be found in usual ductal hyperplastic lesions, including several lesions with multiple events. This implies that usual ductal hyperplastic lesions may be among the earliest lesions within the breast oncogenetic spectrum. PMID- 9865846 TI - Systemic mast cell disease associated with primary ovarian mixed malignant germ cell tumor. AB - A 12-year-old girl with a mixed malignant germ cell tumor of the ovary, treated by surgery and chemotherapy, developed systemic mast cell disease (SMCD) approximately 3 months after chemotherapy. Hematologic malignancies have previously been noted in patients with mediastinal germ cell tumors but this is the first report of a primary ovarian germ cell neoplasm associated with SMCD. PMID- 9865847 TI - Solid tumors and germ cell tumors induce nonneoplastic germ cell proliferations in testes of infants and young children. PMID- 9865848 TI - Clear cell cholangiocarcinoma of the liver is a morphologically distinctive entity. PMID- 9865849 TI - 1898-1998: Camillo Golgi and "the Golgi": one hundred years of terminological clones. AB - Camillo Golgi discovered, in 1898, the cell organelle that has been called, after him, "the Golgi apparatus" or "the Golgi complex." Because of the necessity of saving words in scientific literature, "the Golgi apparatus" is nowadays frequently referred to only as "the Golgi" or used in an adjectival form in combination with a variety of terms (e.g., Golgi vesicle, Golgi area, Golgi traffic, and so forth). We present here a search of the Medline database for the years 1967 through 1997, which demonstrates that the eponym has lost all connection with Golgi's personal identity. In addition, the search indicates that interest in the Golgi apparatus is steadily increasing, as evidenced in the scientific literature, and that Golgi's name is much more frequently used in association with the cell organelle than with any of the other scientific contributions of this scientist, including the Golgi impregnation of nerve cells. PMID- 9865850 TI - The history of the Golgi apparatus in neurones from its discovery in 1898 to electron microscopy. AB - We celebrate in 1998 the centenary of the discovery of the Golgi apparatus. Neurones have played a special role in the history of this cell organelle, primarily because the endocellular apparatus was discovered in nerve cells by Camillo Golgi when he was studying the cerebellum of an owl. In the following years, the apparatus of the nerve cell represented the prototype of this cell component which, however, was soon also detected in many other cell types. During the 1920s and 1930s the original idea of a Golgi endocellular "network" was refused and research was focused on the constituents and function of the organelle, using various tissues and techniques. However, the real existence of the apparatus was seriously questioned until the organelle was finally identified by electron microscopy in the mid-1950s. The studies performed on nerve cells during these decades is briefly reviewed here. PMID- 9865851 TI - Effects of halothane and methoxyflurane on the hypothalamic-pituitary-adrenal axis in rat. AB - Effects of acute exposure (2 h) to either 1.5% halothane or 0.5% methoxyflurane on chemical mediators of the hypothalamic-pituitary-adrenal (HPA) axis were evaluated in male Sprague-Dawley rats immediately after exposure, after the righting reflex (4 h), or 24 h postexposure. Effects of these anesthetics on hippocampal corticotropin releasing factor (CRF) were also evaluated. Methoxyflurane caused significant elevations in pituitary adrenocorticotropin hormone (ACTH)-like immunoreactivities in all three of the experiment's time groups, yet halothane failed to cause the same response immediately after exposure. Serum ACTH-like immunoreactivities were significantly elevated immediately after exposure to both anesthetics, but were not elevated at 4 and 24 h postexposure. Corticosterone (CORT)-like immunoreactivities were significantly elevated by halothane in all experimental groups, and in the 2- and 24-h groups following methoxyflurane exposure. Hippocampal CRF-like immunoreactivities remained unaffected by either anesthetic. Results indicate that a 2-h exposure to either halothane or methoxyflurane results in significant activation of the rat hypothalamic-pituitary-adrenal axis, and that the activation appears to be sustained over a 24-h period. PMID- 9865852 TI - Effects of a lesion in the dorsal raphe nuclei performed during the juvenile period of the female rat, on puberty. AB - The present study examined the effect of thermic lesions on the dorsal raphe nuclei (DRN) of the female rat, performed at various ages during the prepubertal period (21, 24, or 27 days), on puberty, and at first ovulation. In comparison with sham-operated animals, the age of vaginal opening and first vaginal oestrus was delayed in rats with a DRN lesion performed at the end of the infantile period (day 21) (45.6+/-0.94 vs. 89.9+/-1.03, p < 0.05), whereas differences were not observed in animals with lesions performed during the juvenile period (day 24 or 27). The number of ova shed by ovulating animals was greater in those rats with lesions performed on day 24 or 27 (9.7+/-0.4 vs. 7.4+/-0.4; 9.5+/-0.5 vs. 7.7+/-0.4, p < 0.05). Ovarian follicular atresia in these animals was significantly lower than in control and sham-operated ones. On the day of first vaginal oestrus and 48 h after the lesion, the serotonin-hypothalamic concentration decreased in all lesioned animals. Present results support the idea of the participation of the serotoninergic system, arising from the DRN, in the neuroendocrine mechanisms regulating the onset of puberty and the first ovulation, with variations depending on animal maturity. PMID- 9865853 TI - Rate of neuronal fallout in a transsynaptic cerebellar model. AB - Quantitative analyses of transsynaptic granule cell death subsequent to the genetically determined degeneration of Purkinje cells in the cerebellum of pcd/pcd mutant mice show that granule neuron fallout follows a typical mathematical pattern of elemental decay. Biological and theoretical connotations are discussed in light of the empirical observations and a simulation model. PMID- 9865854 TI - Mitogen-activated protein kinase is increased in the limbic structures of the rat brain during the early stages of status epilepticus. AB - Systemic administration of pilocarpine (PILO) in adult rat produces acute limbic seizures leading to status epilepticus. Recent studies have shown the activation of mitogen-activated protein kinase (MAPK) cascades during experimentally induced seizures. MAPK activation may be triggered by glutamatergic stimulation and may play a key role in signal transduction pathways. In the present study, immunocytochemistry was used to analyze the spatiotemporal distribution pattern of the MAPK protein and its active form (A-MAPK) following PILO-induced status epilepticus. MAPK and A-MAPK immunoreactivities exhibited different patterns of distribution in the brain of normal and epileptic rats. The saline-treated rats, as well as the animals that received PILO but did not evolve to status epilepticus, showed a weak but selective MAPK immunoreactivity, detected in the hippocampal pyramidal neurons, dentate gyrus, hilus, CA3, CA1, and entorhinal, piriform, and cingulate cortices. A-MAPK immunoreactivity was instead observed only in neurites of the CA3 and hilus and in cells of the entorhinal and piriform cortices. In PILO-treated rats, between 30 and 60 min after status epilepticus there was an increase of the immunoreactivity to both antibodies, which were differently distributed throughout several structures of the limbic system. The immunostaining showed a slight decrease after 5 h of status epilepticus. However, MAPK and A-MAPK immunopositivities decreased markedly after 12 h of status epilepticus, returning almost to the basal expression. These findings are consistent with a spatial and time-dependent MAPK expression in selected limbic structures, and its activation could represent an initial trigger for neuronal modifications that may take part in the mechanism underlying acute epileptogenesis and in long-lasting neuropathological changes of the PILO model of epilepsy. PMID- 9865855 TI - Modulation of TNF-alpha mRNA production in rat C6 glioma cells by TNF-alpha, IL 1beta, IL-6, and IFN-alpha: in vitro analysis of cytokine-cytokine interactions. AB - Cytokines regulate the expression of other cytokines in the centrally derived rat C6 glioma cell line. However, the modulation of tumor necrosis factor-alpha (TNF alpha, a pivotal proinflammatory cytokine) in C6 cells is unknown. Here we investigated the expression of TNF-alpha mRNA in C6 glioma cells in response to TNF-alpha, interleukin-1beta (IL-1beta), IL-1 receptor antagonist (IL-1Ra), interleukin-6 (IL-6), and interferon-alpha (IFN-alpha). The data show that (1) IL 1beta induced a significant upregulation of TNF-alpha mRNA; (2) the effect of IL 1beta on TNF-alpha mRNA expression was completely blocked by the concomitant application of IL-1Ra, which suggests specificity of IL-1beta action through the IL-1 signaling receptor; (3) no detectable modulation of TNF-alpha mRNA expression was observed with the individual applications of TNF-alpha, IL-6, or IFN-alpha; (4) the concomitant treatments of TNF-alpha + IL-1beta or TNF-alpha + IL-1beta + IL-6 strongly upregulated TNF-alpha mRNA expression, whereas the concomitant application of TNF-alpha + IL-6 or IL-1beta + IL-6 induced a moderate increase; and (5) IFN-alpha significantly attenuated induction of TNF-alpha mRNA by TNF-alpha + IL-1beta + IL-6. Thus, IL-1beta, TNF-alpha and IL-6 interact to upregulate TNF-alpha mRNA expression synergistically, and IFN-alpha acts as an inhibitory cytokine in C6 glioma cells. These findings also suggest that the rat C6 glioma cell line may be used as an in vitro model to characterize cytokine cytokine interactions. PMID- 9865856 TI - Vulnerability to stressor-induced disturbances in self-stimulation from the dorsal and ventral A10 area: differential effects of intraventricular D-Ala2-Met5 enkephalinamide, D-Ala2, N-Me-Phe4, Gly-Ol5-enkephalin, and D-Pen2, D-Pen5 enkephalin administration. AB - D-Ala2-Met5-enkephalinamide (DALA) (1.0 microg/microl) was administered intraventricularly to mice responding for electrical stimulation from the dorsal or ventral aspects of the VTA immediately prior to footshock (Experiment 1). Predictably, footshock reduced self-stimulation from the dorsal but not the ventral VTA immediately, 24, and 168 h following the stressor. Intraventricular DALA administration effected a partial attenuation of stressor-induced self stimulation reductions from the dorsal VTA immediately and 24 h poststressor. Deficits appeared among DALA-Shocked mice responding for brain stimulation from the ventral VTA during comparable test intervals. The long-term depressant influence of footshock on self-stimulation from the dorsal VTA was abolished among DALA-treated mice and DALA-associated reductions in self-stimulation from the ventral A10 region among stressed mice were not evident 1 week later. Administration of D-Ala2, N-Me-Phe4, Gly-Ol5-enkephalin (DAGO) (0.01 microg/microl) or D-Pen2, D-Pen5-enkephalin (DPDPE) (1.0 microg/microl) intraventricularly prior to footshock effected an immediate and a delayed antagonism, respectively, of the stressor on self-stimulation from the dorsal VTA, which persisted for 1 week. Prophylactic administration of 0.001 microg/microl DAGO or 0.01 microg/microl DPDPE prior to the stressor failed to influence self-stimulation from the ventral VTA (Experiment 2). Administration of 0.01 microg/microl DAGO or 1.0 microg/microl DPDPE among mice responding for brain stimulation from the dorsal VTA following footshock produced a weak therapeutic effect immediately poststressor, but effected protracted amelioration of footshock-induced reductions of self-stimulation from the dorsal VTA (Experiment 3). Taken together, mu, delta, and mu-delta activation influenced self-stimulation differentially from the dorsal and ventral VTA according to the temporal order of opioid peptide challenge relative to stressor imposition. These data are discussed with respect to stressors, motivational alterations, and the putative modulating influence of endogenous enkephalin activity in subareas of the VTA. PMID- 9865857 TI - Human working memory capacity is 7+/-2 in a radial maze with distracting interruption: possible implication for neural mechanisms of declarative and implicit long-term memory. AB - Human participants were instructed to walk out along each of the arms of a 15-m in diameter, 8-arm radial maze once and only once. In order to approximate the circumstances under which laboratory rats remember visited sites, our human participants were asked to select arms in an unsystematic order. They scored an average of 7.6 to 7.8 correct choices, even if midway during a trial there was a 5-min interruption filled with a verbal-spatial interfering task (a scavenger hunt) or a 15-min interruption filled with a visuospatial task (a maze-running computer simulation). This finding extends our earlier research with humans in 13 or 17-arm radial mazes under nondelay conditions, in which we also found working memory (WM) capacity for about 7 to 9 places, the same as that of laboratory rats. We discuss earlier findings in other laboratories, showing that rats can successfully bridge long radial maze task interruptions of 5 or 8 h, and we compare our results also to those from studies in which human participants were not discouraged from reducing memory load by responding systematically in radial mazes. Because the radial maze task takes minutes to complete even under nondelay conditions its routine consideration as a working memory task in the animal literature alters the assumptions often made about the duration of WM in the human literature. Accumulating empirical findings about place-memory in humans, nonhuman mammals, and birds suggest it might be productive to reevaluate this theoretical issue with respect to present knowledge about the roles of the hippocampus and other brain structures in declarative memory and in procedural or implicit memory, while considering the hypothesis that some forms of information may exploit long-term memory in parallel with working memory. PMID- 9865859 TI - African trypanosomiasis in the rat alters melatonin secretion and melatonin receptor binding in the suprachiasmatic nucleus. AB - Rats infected with Trypanosoma brucei brucei, a subspecies of the extracellular parasites that cause African sleeping sickness, were examined for disturbances in the circadian rhythms of melatonin secretion (evaluated by determination of the excretion of melatonin in the urine) and the binding of melatonin to its receptor in the suprachiasmatic nuclei of the anterior hypothalamus. In normal and infected rats, Cosinor analysis showed a significant nocturnal peak. The amplitude of this peak was, however, significantly decreased in the infected rats. The peak of melatonin receptor binding in the suprachiasmatic nuclei showed a 4-h phase advance in the infected rats, compared with the controls (0400 and 0800, respectively). These data point to a disturbance in the circadian rhythm of the melatonin-generating systems in the pathogenesis of African sleeping sickness. PMID- 9865858 TI - Endothelium-dependent relaxation to hydrogen peroxide in canine basilar artery: a potential new cerebral dilator mechanism. AB - In prostglandin F2alpha(PGF2alpha)-precontracted isolated canine basilar arterial rings, hydrogen peroxide (H2O2) produced endothelium-dependent relaxations at concentrations of from 4.4 x 10(-7) - approximately 4.4 x 10(-5) M. Removal of extracellular Ca2+ ([Ca2+]0) attenuated the relaxant effects of H2O2. Complete inhibition of H2O2 relaxant action was obtained after buffering intracellular Ca2+ ([Ca2+]i), in the endothelial cells, with 10 microM 1,2-bis (2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM). The H2O2-induced relaxations could be abolished completely by 1200 u/ml catalase and was suppressed significantly by 0.5 microM atropine, 150 microM NG-monomethyl-arginine (L-NMMA), 50 microM NG nitro-L-arginine methyl ester (L-NAME), 1 microM Fe2+, or 5 microM methylene blue. These inhibitory effects of L-NMMA, L-NAME, or atropine could be reversed partly by 50 microM L-arginine. The Fe2+ inhibition of H2O2-stimulated relaxation was reduced significantly by either 1 mM deferoxamine (a Fe2+ chelator) or 100 microM dimethyl sulfoxide (DMSO, a *OH scavenger). Such relaxant effects of H2O2 were enhanced, significantly, by an acetylcholinesterase antagonist, neostigmine. A variety of pharmacological antagonists (of diverse vasodilator agents) could not inhibit the relaxant action of H2O2. Our observations suggest that at suitable pathophysiological concentrations, H2O2 could induce release of an endothelium-derived relaxing factor (EDRF), probably nitric oxide (NO), from endothelial cells of the canine cerebral artery. The H2O2 relaxant effects are clearly Ca2+-dependent, require formation of cyclic guanosine monophosphate (cGMP), and may be associated with release of endogenous acetylcholine (ACh). PMID- 9865860 TI - Daily and seasonal variations in the concentration of melatonin in the human pineal gland. AB - To elucidate whether pineal melatonin secretion is affected by changes in day length, we determined the concentration of melatonin in human pineal glands obtained at autopsy from 66 male subjects, aged 16-84 years over a period of 12 consecutive months. Based on the time of death, a day-night difference in pineal melatonin levels was evident only in the long photoperiod (April-September) with significantly higher melatonin concentrations occurring at night (2200-1000 h). Nighttime values in the long photoperiod were significantly higher than the nighttime values during the short photoperiod (October-March). During the short photoperiod, the data suggested a possible phase-delay in melatonin secretion. Day-night difference was evident in young subjects (30-60 years), but not in elderly subjects (61-84 years). Elderly subjects had lower total melatonin levels (day and night values) although statistically not significant. Therefore, melatonin levels did not decline with age and when the data were analyzed by age there was no significant day-night difference in melatonin levels. These data indicate that the concentration of melatonin in the human pineal is augmented only during the long photoperiod. The results suggest a partial effect of photoperiod on melatonin secretion in humans. This may result from living in an artificial light environment or due to other nonphotic signals involved in generating melatonin rhythm. PMID- 9865861 TI - Intracerebroventricular infusion of CRF increases extracellular concentrations of norepinephrine in the hippocampus and cortex as determined by in vivo voltammetry. AB - Previous studies have indicated that intracerebroventricular (i.c.v.) infusions of corticotropin-releasing factor (CRF) activate locus coeruleus (LC) noradrenergic neurons and increase the metabolism and extracellular concentrations of norepinephrine (NE) in several brain regions, suggesting increased release. To examine the temporal aspects and mechanism of the presumed release of NE, CRF was infused i.c.v. and the oxidation current was recorded using carbon fiber voltammetric electrodes placed in rat hippocampus or cortex. The CRF (1 microg, i.c.v.) caused a significant increase of oxidation current with a delay of approximately 5 min, and a peak at approximately 35 min. Similar responses were observed in the medial prefrontal cortex. The hippocampal response was markedly attenuated when CRF was infused into rats pretreated with DSP-4 to deplete NE, suggesting that the observed changes in current resulted from oxidation of NE. The increase of NE-like current did not occur when 25 microg alpha-helical CRF9-41 (ahCRF) was injected immediately before 1 microg CRF, suggesting that the response was mediated by cerebral CRF-receptors. Subcutaneous pretreatment with the ganglionic blocker, chlorisondamine, at a dose of 3 mg/kg had no effect on the voltammetric response to CRF, but a 6 mg/kg dose completely prevented the response. The beta-adrenoceptor antagonists, S-propranolol (5 mg/kg), nadolol (5 and 10 mg/kg), and timolol (5 mg/kg) attenuated the NE response to i.c.v. CRF to varying degrees. When chlorisondamine (3 microg) or nadolol (5 microg) were given i.c.v. before the CRF, the hippocampal responses were not blocked. These results suggest peripheral actions of ganglionic and beta adrenergic blockers. We conclude that peripheral autonomic mechanisms, and probably both central and peripheral beta-adrenoceptors, contribute to the increased secretion of hippocampal NE in response to i.c.v. CRF. PMID- 9865863 TI - Leishmaniasis: a re-emerging zoonosis. PMID- 9865862 TI - Structural correlates for down-regulation of m1 and m2 muscarinic receptor subtypes. AB - Three chimeric receptors stably expressed in murine fibroblast (B82) cells were used to examine how different parts of the rat muscarinic m1 and m2 receptors contribute to the down-regulation process. The MCH7 chimeric m2 receptor contained a fragment between VIth TM and C-terminal end derived from the m1 receptor. The MCH3 and MCH5 receptors have exchanged N-terminal and third intracellular loop regions of the MCH7 receptor. Fibroblast cells stably expressing individual muscarinic wild type (m1, m2) or chimeric (MCH3, MCH5, or MCH7) receptors were treated with plain medium (control) or medium containing carbachol for 24 h. Receptor density changes were measured by [3H](-)1-N-methyl-3 quinuclidinyl benzilate ([3H](-)MQNB) saturation binding studies. There was a significant loss of receptor density, different for each receptor studied, following carbachol treatment relative to control cells. We related this loss of [3H](-)MQNB binding to the number of amino acids derived from m1 or m2 receptors for each constructed chimera and to the affinity of carbachol to the receptors studied. We demonstrate that: 1) the region from the VIth TMD to the end of C terminal controls the extent of m1 and m2 receptor down-regulation; 2) the overall receptor conformation and the interaction between intracellular portions of the receptor influence the extent of receptor down-regulation; and 3) resistance to down-regulation by carbachol correlates with the affinity of carbachol to the muscarinic receptor construct. PMID- 9865864 TI - Fluconazole for the treatment of onychomycosis: an update. PMID- 9865865 TI - The vital role of the skin in human natural history. PMID- 9865866 TI - Nomenclature for fungus infections. PMID- 9865867 TI - Seborrheic dermatitis or tinea capitis: don't be fooled. PMID- 9865868 TI - Reticulate postinflammatory hyperpigmentation with band-like mucin deposition. AB - BACKGROUND: Mucinoses of the skin are a group of disorders sharing accumulation of mucin in the skin or hair follicles. Postinflammatory hyperpigmentation, with pigmentary incontinence, is due to loss of melanin from epidermal basal cells and its accumulation in dermal macrophages. METHODS: We describe clinicopathologic features of two patients presenting with the association of pigmentary incontinence with an unusual diffuse, band-like dermal deposition of mucin, clinically presenting with reticular pigmented macular lesions. RESULTS: Two patients were observed with asymptomatic, persistent, reticular, pigmented patches located in the flexures, thighs, neck and back. Histology showed melanophages with a diffuse, band-like dermal deposition of mucin, an increased number of fibroblasts, a slight T-cell infiltrate and scattered mast cells. Blood markers of lupus erythematosus were negative. CONCLUSIONS: These findings may draw attention to pigmentary disorders such as lichen planus pigmentosus, erythema dyschromicum perstans, pigmentatio maculos eruptiva idiopathica, dermatopathia pigmentosa reticularis, prurigo pigmentosa and frictional melanosis. None of these entities, however, includes mucin deposition among its microscopic features. Macules were not preceded by erythema or any other lesions. We suggest that our cases could belong to group II of Rongioletti and Rebora's classification, i.e. they could be cases of secondary mucin deposition in postinflammatory hyperpigmentation, possibly in an unusual form of lichen planus pigmentosus or, less likely, frictional melanosis. PMID- 9865869 TI - Drugs causing fixed eruptions: a study of 450 cases. AB - BACKGROUND: Drug eruptions are among the most common cutaneous disorders encountered by the dermatologist. Some drug eruptions, although trivial, may cause cosmetic embarrassment and fixed drug eruption (FDE) is one of them. The diagnostic hallmark is its recurrence at previously affected sites. OBJECTIVE: We evaluated 450 FDE patients to determine the causative drugs. RESULTS: The ratio of men to women was 1:1.1. The main presentation of FDE was circular hyperpigmented lesion. Less commonly FDE presented as: nonpigmenting erythema, urticaria, dermatitis, periorbital or generalized hypermelanosis. Occasionally FDE mimicked lichen planus, erythema multiforme, Stevens-Johnson syndrome, paronychia, cheilitis, psoriasis, housewife's dermatitis, melasma, lichen planus actinicus, discoid lupus erythematosus, erythema annulare centrifugum, pemphigus vulgaris, chilblains, pityriasis rosea and vulval or perianal hypermelanosis. Cotrimoxazole was the most common cause of FDE. Other drugs incriminated were tetracycline, metamizole, phenylbutazone, paracetamol, acetylsalicylic acid, mefenamic acid, metronidazole, tinidazole, chlormezanone, amoxycillin, ampicillin, erythromycin, belladonna, griseofulvin, phenobarbitone, diclofenac sodium, indomethacin, ibuprofen, diflunisal, pyrantel pamoate, clindamycin, allopurinol, orphenadrine, and albendazole. CONCLUSIONS: Cotrimoxazole was the most common cause of FDE, whereas FDE with diclofenac sodium, pyrantel pamoate, clindamycin, and albendazole were reported for the first time. FDE may have multiform presentations. PMID- 9865870 TI - A randomized, controlled and blinded study of papulopustular lesions in Turkish Behcet's patients. AB - BACKGROUND: Papulopustular lesions (PPL), the commonest presentation of skin lesions in Behcet's disease (BD) are cutaneous, sterile folliculitis or acne-like lesions on erythematous base. Our purpose was to determine the true frequency and anatomic location of the PPL and compare this with controls. We also sought to determine whether or not there was any relationship between PPL and either disease activity or other manifestations of BD. METHODS: Fifty patients with BD, diagnosed according to the criteria of the International Study Group for Behcet's Disease, were enrolled in the study. The control group consisted of 100 patients with other dermatologic diseases (21 acne and 79 non-acne patients), selected randomly. A dermatologist counted the lesions, in a blind protocol, on seven anatomic locations: scalp, face, neck, trunk, upper and lower extremities and genitalia. RESULTS: The frequency of PPL in patients with BD was 96% and the most common location was the trunk, whereas in the control group the frequency was 89% and the most common location was the face. In acne and non-acne patients, the frequency was 100% and 86.1% respectively. The total mean number, and mean numbers of PPL on the location of trunk, upper and lower extremities, and genitalia were higher in patients with BD than in controls. When the PPL in BD patients with a positive pathergy test was compared with that in patients with a negative pathergy test, the difference was significantly higher. CONCLUSIONS: Our results indicate that PPL appear to be non-specific. In the diagnosis of BD the mean number and anatomic location of the lesions are of more importance than the frequency. PMID- 9865871 TI - Cutaneous leishmaniasis in Multan, Pakistan. AB - BACKGROUND: Cutaneous leishmaniasis (CL) is endemic in certain areas of Pakistan. This study was carried out to determine the incidence and clinical pattern of the disease in Pakistan. METHODS: CL patients presenting in the out-patient clinic of the Department of Dermatology, Nishtar Hospital, Multan from January 1995 to June 1997 were included in the study. The patients were diagnosed clinically and confirmed by laboratory demonstration of the parasite in a Giemsa-stained smear from the lesion. All important clinical details were recorded on specially designed forms. RESULTS: Three hundred and five patients with 681 lesions were seen during 30 months. All the lesions were of the dry type. Most of the lesions (97%) were present on exposed areas of the body. One hundred and thirty-four (44%) patients had one and 75 (24%) had two lesions. Eight five per cent of the sufferers were less than 30 years of age. The disease was slightly more preponderant in males (60%). Most cases presented during winter. CONCLUSIONS: Lack of wet type lesions may be due to complete absence of Leishmania major in this area. This is in contrast to other areas of Pakistan where both dry and wet types of CL are present. PMID- 9865872 TI - Leishmaniasis recidiva cutis in New World cutaneous leishmaniasis. AB - BACKGROUND: Leishmaniasis recidiva cutis (LRC) is rare in New World leishmaniasis. Only seven cases have been reported so far. MATERIALS AND METHODS: Four cases are reported here. Parasite diagnosis was performed by classical methods of touch preparations, histopathologic sections, and cultures. In addition, the detection of parasite DNA by the polymerase chain reaction (PCR) was performed in all cases. RESULTS: Parasites were detected by at least one of the classical methods in all primary lesions; however, only the PCR was positive in the recidivant lesions. DISCUSSION: LRC cases most likely represent a reactivation of an initial infection, probably due to the persistence of parasites in scarred tissue. Although lupoid leishmaniasis (LL) has been used as a synonym of LRC, a clear difference between LRC and LL can be defined as LL is the initial clinical presentation while LRC is a recurrent lesion. CONCLUSIONS: The results indicate that it is not appropriate to use these two denominations as synonyms. The designation of LRC should be maintained in order to define recidives occurring at the border of an old scar of cutaneous leishmaniasis, avoiding the confusion with the lupoid form of the disease. PMID- 9865873 TI - Evidence-based dermatologic out-patient treatment. AB - OBJECTIVE: To determine the evidence base for routine therapeutic decisions in dermatologic out-patients. DESIGN: A retrospective review of a random sample of primary therapy and literature. SETTING: University hospital, dermatologic out patient clinic in Copenhagen MATERIAL: A random sample of the case notes from 115 out-patients. METHOD: The evidence base of therapy prescribed when the diagnosis was ascertained was studied in literature searches in MEDLINE and EMBASE. Evidence was structured into primary evidence consisting of randomized controlled trials, and secondary evidence consisting of follow-up studies or the application of trial results between diseases with pathogenic or clinical similarities, e.g. atopic and seborrheic dermatitis. RESULTS: Randomized controlled trials could be found describing 38% (95% confidence interval: 30-47) of all treatments. Secondary evidence was found for 33% (24-41), while no evidence was found for 23% (16-31) of the given treatments. CONCLUSIONS: Approximately three-quarters of dermatologic out-patient therapy is based on scientific evidence ranging from randomized controlled trials to logical deduction from analogous clinical situations. The proportion of evidence-based medicine in dermatologic therapy therefore appears to be comparable with that of internal medicine and may thus be above expectations. PMID- 9865874 TI - Clinical dermatologic education and the diagnostic acumen of medical students and primary care residents. PMID- 9865875 TI - Lichen planus or lichenoid tattoo reaction? PMID- 9865876 TI - Ulcerative lichen planus of the sole with rheumatoid arthritis. PMID- 9865877 TI - Lupus miliaris disseminatus faciei evoked during pregnancy in a patient with cutaneous lupus erythematosus. PMID- 9865878 TI - Simultaneous occurrence of tuberculous gumma, tuberculosis verrucosis cutis, and lichen scrofulosorum. PMID- 9865879 TI - Fluconazole in the treatment of tinea capitis. PMID- 9865880 TI - Terbinafine 1% cream vs. bifonazole 1% cream in the treatment of tinea cruris. PMID- 9865881 TI - Dermatofibrosarcoma protuberans in two distant locations. PMID- 9865882 TI - Neural interfaces for regenerated nerve stimulation and recording. AB - A class of implantable, regeneration-type neural interfaces (NI's) for mammalian peripheral nerve recording and stimulation were developed using different fabrication processes and integrating purposely designed components. A typical NI comprises three main components: 1) a microfabricated silicon die incorporating a microelectrode array on multiple through-holes, 2) a polymer guidance channel housing the die, and 3) a flexible flat cable connecting the die to an external electronic circuitry. The design and fabrication of the NI's were aimed at achieving long term, reliable implants by taking into careful account the biological, electrical, and mechanical requirements of the specific implant site. Different versions of the NI were fabricated and implanted between the severed ends of the sciatic nerve in a mammalian animal model (rabbit). Morphological and histological evidence showed that nerves regenerated through the NI's and electrophysiological results demonstrated the recovery of electrical functionality. Moreover, the NI's allowed stimulation of the regenerated nerve producing a visible leg/foot contraction. The NI's presented in this paper are being further improved in the authors' laboratories with the ultimate goal of allowing the control of nerve motor and sensory functions in future prosthetic devices. PMID- 9865883 TI - Stability of the input-output properties of chronically implanted multiple contact nerve cuff stimulating electrodes. AB - The objective of this investigation was to measure the input-output (I-O) properties of chronically implanted nerve cuff electrodes. Silicone rubber spiral nerve cuff electrodes, containing 12 individual platinum electrode contacts, were implanted on the sciatic nerve of seven adult cats for 28-34 weeks. Measurements of the torque generated at the ankle joint by electrical stimulation of the sciatic nerve were made every 1-2 weeks for the first 6 weeks post-implant and every 3-5 weeks between 6 weeks and 32 weeks post-implant. In three implants the percutaneous lead cable was irreparably damaged by the animal within 4 weeks after implant and further testing was not possible. One additional lead cable was irreparably damaged by the animal at 17 weeks post-implant. The three remaining implants functioned for 28, 31, and 32 weeks. Input-output curves of ankle joint torque as a function of stimulus current amplitude were repeatable within an experimental session, but there were changes in I-O curves between sessions. The degree of variability in I-O properties differed between implants and between different contacts within the same implant. After 8 weeks, the session to session changes in the stimulus amplitude required to generate 50% of the maximum torque (I50) were smaller (15+/-19%, mean +/- s.d.) than the changes in I50 measured between 1 week and 8 weeks post-implant (34+/-42%). Furthermore, the I-O properties were more stable across changes in limb position in the late post implant period than in acutely implanted cuff electrodes. These results suggest that tissue encapsulation acted to stabilize chronically implanted cuff electrodes. Electrode movement relative to the nerve, de- and regeneration of nerve fibers, and the inability to precisely reproduce limb position in the measurement apparatus all may have contributed to the variability in I-O properties. PMID- 9865884 TI - Penile erection produced by microstimulation of the sacral spinal cord of the cat. AB - The sacral neural pathways mediating penile erection in the cat were studied by measuring the change in cavernous sinus pressure (CSP) elicited by stimulation of the sacral ventral roots or by microstimulation of the sacral spinal cord. Ventral root stimulation revealed that the S1 segment rather than S2 and S3 spinal segments could evoke the largest CSP responses. Microstimulation in the S1 spinal cord elicited large CSP responses but small or no bladder contractions. Maximal CSP responses were evoked by microstimulation in the middle of the S1 ventral horn, 1.6-2.8 mm below the cord surface and midway between the midline and the lateral edge of the gray matter. The area was 200-400 microm wide (medial to lateral) and extended 1-2 mm in the rostrocaudal direction. Maximal CSP responses to spinal cord microstimulation were elicited by stimulus intensities of 50-150 microA, at a pulse width of 0.2 ms and at frequencies of 3040 Hz and occurred after delay of 8-40 s. This study suggests that focal microstimulation of the sacral spinal cord might be useful in eliciting penile erectile activity in patients with spinal cord injury. PMID- 9865885 TI - Laparoscopic placement of electrodes for diaphragm pacing using stimulation to locate the phrenic nerve motor points. AB - Laparoscopic mapping of the phrenic nerve motor points using test stimulation was conducted for the implant of epimysial electrodes for diaphragm pacing in dogs. Both visual assessment of muscle activation and measurements of recruitment were useful for identifying an implant location resulting in a mean electrode placement approximately 14 mm from the phrenic nerve motor points in 16 dogs. Postmortem analysis of the stimulus test site locations and corresponding recruitment curves suggested that the phrenic nerve motor points could be predicted during the laparoscopic procedure to within 4.5 mm of the anatomical motor point. PMID- 9865886 TI - Accessing the tonotopic organization of the ventral cochlear nucleus by intranuclear microstimulation. AB - This study is part of a program to develop an auditory prosthesis for the profoundly deaf, based on multichannel microstimulation in the cochlear nucleus. The functionality of such a device is dependent on its ability to access the tonotopic axis of the human ventral cochlear nucleus in an orderly fashion. In these studies, we utilized the homologies between the human and feline ventral cochlear nuclei and the known tonotopic organization of the central nucleus of the inferior colliculus (IC). In anesthetized cats, stimuli were delivered to three or four locations along the dorsal-to-ventral axis of the posteroventral cochlear nucleus (PVCN), and for each stimulus location, we recorded the multiunit neuronal activity and the field potentials at 20 or more locations along the dorsolateral-ventromedial (tonotopic) axis of the IC. The current source-sink density (CSD), which delimits regions of neuronal activity, was computed from the sequence of field potentials recorded along this axis. The multiunit activity and the CSD analysis both showed that the tonotopic organization of the PVCN can be accessed in an orderly manner by intranuclear microstimulation in several regions of the PVCN, using the range of stimulus pulse amplitudes that have been shown in previous studies to be noninjurious during prolonged intranuclear microstimulation via chronically implanted microelectrodes. We discuss the applicability of these findings to the design of clinical auditory prostheses for implantation into the human cochlear nucleus. PMID- 9865887 TI - EMG pattern recognition based on artificial intelligence techniques. AB - This paper presents an electromyographic (EMG) pattern recognition method to identify motion commands for the control of a prosthetic arm by evidence accumulation based on artificial intelligence with multiple parameters. The integral absolute value, variance, autoregressive (AR) model coefficients, linear cepstrum coefficients, and adaptive cepstrum vector are extracted as feature parameters from several time segments of EMG signals. Pattern recognition is carried out through the evidence accumulation procedure using the distances measured with reference parameters. A fuzzy mapping function is designed to transform the distances for the application of the evidence accumulation method. Results are presented to support the feasibility of the suggested approach for EMG pattern recognition. PMID- 9865888 TI - Performance statistics of a head-operated force-reflecting rehabilitation robot system. AB - Researchers in the rehabilitation engineering community have been designing and developing a variety of passive/active devices to help persons with limited upper extremity function to perform essential daily manipulations. Devices range from low-end tools such as head/mouth sticks to sophisticated robots using vision and speech input. While almost all of the high-end equipment developed to date relies on visual feedback alone to guide the user providing no tactile or proprioceptive cues, the "low-tech" head/mouth sticks deliver better "feel," because of the inherent force feedback through physical contact with the user's body. However, the disadvantage of a conventional head/mouth stick is that it can only function in a limited workspace and the performance is limited by the user's strength. It therefore seems reasonable to attempt to develop a system that exploits the advantages of the two approaches: the power and flexibility of robotic systems with the sensory feedback of a headstick. The system presented in this paper reflects the design philosophy stated above. This system contains a pair of master-slave robots with the master being operated by the user's head and the slave acting as a telestick. Described in this paper are the design, control strategies, implementation and performance evaluation of the head-controlled force-reflecting telestick system. PMID- 9865889 TI - Optimal character arrangements for ambiguous keyboards. AB - Many persons with disabilities lack the fine motor coordination necessary to operate traditional keyboards. For these individuals, ambiguous (or reduced) keyboards offer an alternative access method. By placing multiple characters on each key, the size and accessibility of the individual keys can be enhanced without requiring a larger keyboard. Using statistical disambiguation algorithms to automatically interpret each keystroke, these systems can approach the keystroke efficiency (keystrokes per character) of conventional keyboards. Since the placement of characters on each key determines the effectiveness of these algorithms, several methods of optimizing keyboard arrangements have previously been proposed. This paper presents a new method for optimizing an arbitrary set of N characters over a collection of M keys. While earlier efforts relied upon approximations of keystroke efficiency, the proposed approach optimizes the arrangement under this exact performance measure. Applied to the canonical 26 characters on a nine-key "telephone keypad" problem, this method provides an improvement in efficiency of 2.5 percentage points over previously established layouts. Using only a minimum of calculations, the proposed technique operates quickly and efficiently, deriving optimal arrangements in a matter of seconds using a personal computer. The flexible method is applicable to arbitrary disambiguation algorithms, character sets, and languages. PMID- 9865890 TI - A transducer for the measurement of finger joint moments. AB - A device capable of simultaneously measuring the isometric moments generated about the metacarpophalangeal (MP), proximal interphalangeal (PIP), and distal interphalangeal (DIP) joints of all four fingers has been developed. The design utilizes a four-bar linkage to transmit moments, but not forces, to the device. This linkage allows the same device to fit a wide range of hand sizes without recalibration. The device was constructed out of aluminum bars which are strapped to each joint segment and to the back of the hand. Strain gauges mounted to the aluminum bars measure the bending moment on the device, which is directly related to the moment applied about the joint center of rotation. Because of the unique design of the device, it is not necessary to have accurate measurements of the joint center of rotation in order to get accurate moment information. A single device is capable of generating independent measurement of MP extension/flexion, PIP extension/flexion, and DIP extension/flexion. Four of these devices can be used to make simultaneous measurements of all the moments generated by all four fingers. The device also acts as a splint, allowing each joint to be positioned and locked at any angle through the range of motion of the joint. The device is accurate to within +/- 5.6% of each reading for moments from 10 N x cm to 100 N x cm and within +/- 2.0 N x cm for moments of 10 N x cm or less. If the device configuration is constrained, the accuracy can be improved to +/- 0.8% of full scale (100 N x cm) and +/- 0.21 N x cm for moments of 10 N x cm or less. The device can measure both flexion and extension moments up to 100 N x cm, and can allow the joints to be fixed at any angle from approximately 10 to 80 degrees. PMID- 9865891 TI - Curriculum for the 21st century--or is it the 21-year curriculum? PMID- 9865892 TI - The Cloutterbuck Minimum Data Matrix: a teaching mechanism for the new millennium. AB - The demands of the United States' rapidly changing health care system and society signal the need to design and implement health professions programs which prepare students for practice within a health care paradigm that will be very different from the one existing today and for work with an increasingly diverse and independent consumer population. The Cloutterbuck Minimum Data Matrix (CMDM) is a teaching mechanism designed to generate a comprehensive base of consumer information prerequisite to the indepth level of critical analysis and synthesis needed to produce quality health care outcomes in the 21st century. The CMDM assumes an interdisciplinary perspective and educates students to recognize and incorporate consumer diversity into strategies for care. PMID- 9865893 TI - Teaching complementary healing therapies to nurses. PMID- 9865894 TI - Culture and loss: a project of self-reflection. PMID- 9865895 TI - Shift report: a time for learning. AB - Shift report provides an opportunity for professional communication that supports role socialization and development of a cohesive group process in the health care system. In this learning project, nursing students were able to practice professional behaviors, prioritize information, and reduce material to the boundaries of a work setting without relinquishing the holistic approach to the client. Critical thinking, direct application, self-analysis, peer critiques, and problem-solving processes were used by the student groups which will assist in their role transition to professional nursing practice. PMID- 9865896 TI - Levels of stress and academic performance in baccalaureate nursing students. PMID- 9865897 TI - Competencies of liberal education in post-RN baccalaureate students: a longitudinal study. AB - The purpose of this longitudinal study was to examine the relationship among liberal education competencies as demonstrated in the professional and personal lives of post-RN students and time spent in a BScN program. The six competencies identified by Dressel (1979) and used by Bottoms (1988) to define the liberally educated person provided the framework for this study. The Dressel competencies have been validated in the literature as being representative of the universe of competencies that are anticipated outcomes of baccalaureate nursing education. Results indicated that after 5 years of part-time study at the baccalaureate level, significant differences were found in the competencies of liberal education as demonstrated in the professional and personal lives of post-RN learners. The competencies as demonstrated in the professional lives of post-RN learners developed consistently over time, while the development of competencies as demonstrated in the personal lives of learners lagged behind. The study contributes to the small but growing body of literature which explores the impact of a liberal education on post-RN learners. PMID- 9865898 TI - Infertility education in baccalaureate schools of nursing. AB - The increasing number of couples who enter the current health care system for assistance with fertility needs warrants an examination of the ways nurse educators are preparing nurse generalists to meet those needs. A survey was conducted of baccalaureate nursing programs to determine what, if any, information was being presented on infertility. Analysis of data with a 50% return rate indicated most schools of nursing in this sample are in fact including some information on infertility in their curricula. What is interesting to note is only slightly more than half of respondents included information on the emotional and psychosocial issues of infertility, despite the fact that the greatest cost of infertility is the emotional one. One of the major concerns of infertile couples is health care providers, including nurses, do not seem to understand what they are experiencing. How do nurses obtain such understanding if they are not prepared in their educational program to do so? PMID- 9865899 TI - Tobacco-related instruction in undergraduate nursing education in Illinois. AB - Tobacco use is responsible for more deaths in the United States than any other factor. Nurses are in a unique position to convey life-saving messages to clients regarding tobacco use. To gauge the type and extent of tobacco-related background knowledge acquired by nurses in the course of their education, the Nurses' Committee of the Illinois Division of the American Cancer Society (ACS) surveyed 70 nursing programs in the state of Illinois. The number of lecture hours spent on tobacco-related issues was greater in LPN programs than in either associate or baccalaureate degree programs, and instruction was scattered throughout the curriculum of each program. Most schools reported heavy reliance on adult medical surgical textbooks to convey tobacco-related content. The most recent editions of the textbooks used by the schools were reviewed, and they also were found to adopt a scattered approach, with a disappointing lack of depth regarding the hazards of tobacco. It is recommended a single course be identified as responsible for relaying tobacco-related content and information supplied by general medical-surgical textbooks be supplemented by materials drawn from other sources. PMID- 9865900 TI - In defense of pediatric nursing undergraduate education. PMID- 9865901 TI - Why undertake higher degrees in nursing? PMID- 9865902 TI - In breast cancer, amplification of the steroid receptor coactivator gene AIB1 is correlated with estrogen and progesterone receptor positivity. AB - The AIB1 gene was isolated upon microdissection of the homogeneously staining regions observed in breast cancer cell lines. It was subsequently shown to map at a region at 20q12 that is frequently amplified in breast tumors. In a screen of breast tumor cell lines, of all the genes mapping to the region, AIB1 appeared to be the most consistently amplified and overexpressed. AIB1 shares homology with the SRC-1 family of nuclear receptor coactivators. It was found to interact in a ligand-dependent manner with the estrogen receptor (ER) and to result in increased levels of estrogen-dependent transcription. These properties could be of important biological significance in breast and ovarian cancerigenesis, and we were, therefore, interested in determining whether the amplification of the AIB1 gene was associated with a particular phenotype or subgroup in these tumors. We tested a population of 1157 breast and 122 ovarian tumors in which DNA amplification had been determined previously at 15 chromosomal locations. Amplification of the AIB1 gene was observed in 4.8% of breast cancers and 7.4% of ovarian cancers. In breast tumors, AIB1 was correlated with ER and progesterone receptor positivity, as well as with tumor size. Correlation was also observed with the amplification of MDM2 and FGFR1 genes, but interestingly, no correlation was found with the amplification of CCND1, which is known to be strongly associated with ER. Furthermore, analyzing at 20q12-q13 range, we show the existence of three amplification cores, represented by AIB3/AIB4, AIB1, and RMC20C001. AIB1 and CCND1 amplifications may, thus, represent two different subsets of ER-positive breast tumors. PMID- 9865903 TI - Comparison of features of human breast cancer cell lines and their corresponding tumors. AB - Although human tumor-derived cell lines play an important role in the investigation of cancer biology and genetics, there is no comprehensive study comparing tumor cell line properties with those of the individual tumors from which they were derived. We compared the properties of a series of 18 human breast cancer cell lines that were cultured for a median period of 25 months (range, 9-60 months) and their corresponding archival tumor tissues. We compared morphological characteristics, ploidy, and immunohistochemical expression of estrogen receptors, progesterone receptors, and HER2/neu and p53 proteins. For 17 of these cases, we also tested for allelic losses at 18 chromosomal regions frequently deleted in breast tumors using 51 polymorphic microsatellite markers, and we determined the TP53 gene mutation status in exons 5 to 10. There was an excellent correlation between the breast tumor cell lines and their corresponding tumor tissues for morphological features (100%); presence of aneuploidy (87%); immunohistochemical expression of estrogen receptors (87%), progesterone receptors (73%), and HER2/neu (93%) and p53 proteins (100%); allelic loss at all of the chromosomal regions analyzed (82-100% concordance); and TP53 gene mutations (75%). The same parental allele was lost in 279 (99%) of 281 of the comparisons of allelic losses. The fractional allelic loss indices (a reflection of the total allelic loss) of the cell lines and their corresponding tumor tissues were identical or similar in 15 (88%) of 17 paired comparisons. Although our previous studies (A. Gazdar et al., Int. J. Cancer, in press) indicated that only a subset of primary breast carcinomas that have several features indicative of advanced tumors with poor prognosis can be successfully cultured, the cell lines retain the properties of their parental tumors for lengthy culture periods and, thus, provide suitable model systems for biomedical studies. PMID- 9865904 TI - Basic fibroblast growth factor receptors and their prognostic value in human breast cancer. AB - We performed a saturation binding study with 125I-labeled FGF (fibroblast growth factor)-2 in a nonselected series of 250 human primary breast cancers. Two hundred twenty-five breast cancer biopsies possessed bFGFR (basic FGF receptor). The median dissociation constant was 0.35 nM (range, 0.014-1.9), and the median concentration was 1126 fmol/mg protein (range, 49-7328). FGFR-1 was localized, using a specific monoclonal antibody, in cancerous cells and in epithelial cells in normal breast or in benign tumors. In all of the tissues studied, light stromal cell staining was also observed. Thus, the localization of FGFR-1 in carcinoma cells supports the hypothesis that an important part of FGF-2 binding reflects binding to FGFR-1. bFGFR concentrations were positively correlated to estrogen receptor and progesterone receptor levels. Cox univariate analyses showed that the bFGFR (> or = upper quartile) was associated to longer relapse free survival [P = 0.004; RR (risk ratio), 0.46] and overall survival (P = 0.001; RR, 0.35); age, estrogen receptor levels, progesterone receptor levels, node involvement, tumor diameter, and histoprognostic grading were prognostic, also. In Cox multivariate analyses, only the bFGFR, age, node involvement, and histoprognostic grading were prognostic factors; the bFGFR was associated with longer relapse-free survival (P = 0.03; RR, 0.4) and overall survival (P = 0.009; RR, 0.3). The present study confirms that FGF could be an important regulator of human breast cancer growth and that patients with a high level of bFGFR had a better prognosis. PMID- 9865905 TI - Relationship between paclitaxel activity and pathobiology of human solid tumors. AB - We previously reported the pharmacodynamics of antiproliferative and apoptotic effects of paclitaxel in histocultures of bladder, breast, head and neck, ovarian, and prostate tumors obtained from patients. This study examined the relationship between paclitaxel pharmacodynamics and tumor pathobiological parameters [i.e., stage, grade, proliferation status, expression of P glycoprotein (Pgp), p53, and Bcl-2]. Pgp, p53, and Bcl-2 proteins were detected by immunohistochemical methods. The drug sensitivity rank order of the five tumor types is as follows: prostate > or = head and neck = bladder > breast > ovarian for the antiproliferative effect and breast = ovarian = head and neck > prostate = bladder for the apoptotic effect. When the pathobiological parameters were considered as single parameters, the antiproliferative effect was inversely correlated with tumor stage, grade, labeling index (LI), and expression of Pgp, p53, and Bcl-2 (P < 0.05 in all cases). The apoptotic effect was positively correlated with Pgp expression, LI, and tumor grade (P < 0.01) but was not related to tumor stage and expression of p53 and Bcl-2 (P > 0.2). Results of multivariate analysis indicated that the maximum antiproliferative effect was best predicted by the combination of tumor stage and expression of Pgp and p53 (inverse correlation) and that the maximum apoptotic effect was best predicted by the combination of tumor LI and Pgp expression (positive correlation). In summary, these results indicate that the two major effects of paclitaxel in human solid tumors, i.e., antiproliferation and apoptosis, correlate with different tumor properties. The second finding that drug-induced apoptosis was equal or higher in tumors that expressed Pgp, p53, and Bcl-2 compared to tumors that did not express these proteins supports the use of paclitaxel in treating Pgp-, p53- and Bcl-2-positive tumors. PMID- 9865906 TI - Mouse-human chimeric anti-epidermal growth factor receptor antibody C225 inhibits the growth of human renal cell carcinoma xenografts in nude mice. AB - The epidermal growth factor (EGF) receptor and its ligand transforming growth factor-alpha (TGF-alpha) are overexpressed in human renal cell carcinoma (RCC). The chimeric anti-EGF receptor monoclonal antibody C225 was used to determine the effects of blocking the EGF receptor on RCC growth both in vitro and in vivo. A panel of RCC cell lines all tested positive at various levels for EGF receptor cell surface expression. C225 inhibited DNA synthesis of cultured A498, Caki-1, SK-RC-4, SK-RC-29, and SW839 cells in a dose-dependent manner, ranging from 20 to 45% inhibition compared with untreated controls. C225 also inhibited exogenous ligand-stimulated tyrosine phosphorylation of EGF receptor on RCC cells. The antitumor effects of C225 on RCC tumor growth were evaluated in ascites, s.c., and orthotopic RCC xenograft models. Mice treated with C225 in a Caki-1 ascites xenograft model showed a significant increase in survival (P = 0.002). All control mice died with ascites tumors by week 9, whereas >70% of C225-treated mice survived beyond 12 weeks. C225 also inhibited the growth of s.c. SK-RC-29 tumors in a dose-dependent manner. Mice treated with C225 (1 mg/dose) displayed a significant decrease in tumor volume compared with mice treated with control antibody (P < 0.05) or vehicle alone (P < 0.01). Lastly, C225 inhibited the growth and metastasis of RCC tumors growing orthotopically in the renal subcapsule of nude mice. Histological examination of RCC tumors from mice treated with C225 showed a substantial decrease in proliferating cell nuclear antigen staining and an increase in tumor cell apoptosis. These data suggest that C225 affects growth of RCC tumors by inhibiting EGF receptor-dependent proliferation and demonstrate the potential for therapeutic application of C225 in the treatment of human renal cancer. PMID- 9865908 TI - Effect of weight loss and the inflammatory response on leptin concentrations in gastrointestinal cancer patients. AB - Animal research suggests that leptin may have an important role in the regulation of energy balance. The role of leptin in the progressive involuntary weight loss associated with cancer in humans is of considerable interest. However, such studies are limited. In this study, we compared circulating leptin concentrations in gastrointestinal cancer patients and weight loss (n = 27) with those of healthy subjects (n = 27). The effect of the presence of an inflammatory response on leptin concentrations was also examined. There were significantly lower leptin concentrations in male (median, 2.4 microg/liter; range, <0.5-6.0 microg/liter) and female (median, 3.4 microg/liter; range, <0.5-9.8 microg/liter) cancer patients than there were in male (median, 6.5 microg/liter; range, 3.1-10.9 microg/liter) and female (median, 18.7 microg/liter; range, 8.0-31.5 mcirog/liter) healthy subjects (P < 0.001). However, the leptin concentrations in both patients and normal subjects were related to the predicted percentage of body fat (r = 0.731; P < 0.001). Circulating leptin concentrations in the cancer patients were not altered by the presence of an inflammatory response. These results suggest that cancer anorexia/cachexia is not due to a simple dysregulation of leptin production. PMID- 9865907 TI - Calphostin C triggers calcium-dependent apoptosis in human acute lymphoblastic leukemia cells. AB - Recent studies have demonstrated that the naturally occurring perylenequinone antibiotic calphostin C is a potent inhibitor of protein kinase C and can induce apoptosis in some tumor cell lines by an as yet unknown mechanism. Here we demonstrate that calphostin C induces dose-dependent apoptosis in DT40 chicken lymphoma B-cells, and targeted disruption of lyn, syk, btk, PLCgamma2, or IP3R genes does not prevent or attenuate its cytotoxicity. In our study, calphostin C also induced rapid apoptosis in human acute lymphoblastic leukemia (ALL) cell lines ALL-1 (BCR-ABL+ pre-pre-B ALL), RS4;11 (MLL-AF4+ pro-B ALL), NALM-6 (pre-B ALL), DAUDI (Burkitt's/B-cell ALL), MOLT-3 (T-ALL), and JURKAT (T-ALL), whereas other potent PKC inhibitors did not. In biochemical studies, calphostin C was discovered to induce rapid calcium mobilization from intracellular stores of ALL cell lines, and its cytotoxicity against ALL cell lines was well correlated with the magnitude of this calcium signal. Calphostin C-induced apoptosis was markedly suppressed by BAPTA/AM, a cell-permeable Ca2+ chelator as well as NiCl2, an inhibitor of Ca2+/Mg2+-dependent endonucleases. Inhibition of the Ca2+/calmodulin dependent phosphatase calcineurin with perfluoreperazine dimadeate (a calmodulin antagonist) or cyclosporin A (a specific inhibitor of calcineurin) also reduced the magnitude of calphostin C-induced apoptosis in ALL cell lines. Calphostin C was capable of inducing calcium mobilization and apoptosis in freshly obtained primary leukemic cells from children with ALL. Taken together, our results provide unprecedented evidence that calphostin C triggers a Ca2+-dependent apoptotic signal in human ALL cells. PMID- 9865909 TI - High-dose trimetrexate and minimal-dose leucovorin: a case for selective protection? AB - Our objective was to find the minimum dose of leucovorin (LV; 5 formyltetrahydrofolate) needed to potentially provide selective protection of normal tissue in patients with tumors resistant to methotrexate (MTX) by virtue of transport during prolonged therapy with high-dose trimetrexate (TMTX). Based upon the known daily requirement for folate, that tumors are often resistant to methotrexate via a transport-based mechanism, and that large doses of trimetrexate can be given with large doses of leucovorin for the treatment of patients with Pneumocystis carinii, a protocol was designed to find the minimum LV dose required to allow the administration of large doses of TMTX. Patients were treated in 28-day cycles consisting of 14 consecutive days of oral TMTX (45 mg/m2 every 12 h), followed by 14 days of rest. The dose of concurrent LV was started at 5 mg/m2 twice daily. Cohorts of patients received successive half doses of LV so long as three consecutive patients had less than or equal to grade 3 toxicity. Ten patients received 29 courses of therapy. The most common toxicities encountered were thrombocytopenia (38%), mucositis (14%), and neutropenia (10%). At a LV dose of 2.5 mg/m2, toxicities were consistently limited to less than or equal to grade 3 and only one episode of grade 4 hematological toxicity. Although there was marked interpatient variability, the minimally effective LV dose for selective protection seems to be 2.5 mg/m2. If tumors are resistant to methotrexate because of decreased transport of drug (and also folate), then the same pharmacological principle used to develop TMTX/LV for the treatment of P. carinii may be applied to treatment of some patients with cancer. PMID- 9865910 TI - Coexpression of cholesterol sulfate and cytokeratin as tumor markers in well differentiated squamous cell carcinoma of the human uterine cervix. AB - The expression of cholesterol sulfate (CS) is known to increase during squamous differentiation of keratinocytes and to activate the epsilon, eta, and zeta forms of protein kinase C as a signal transduction molecule for the subsequent expression of transglutaminase-1 (TG-1) and cytokeratins. To gain further insight into the regulation of cellular differentiation and tumorigenesis by CS, we examined the concentration and the potential for synthesis of CS in seven and four surgical specimens from human ovarian and uterine cervical cancer patients, respectively, and eight cell lines established from human uterine cervical cancer patients and compared them for the rate of expression of cytokeratin. CS was present in all of the uterine cervical cancer tissue specimens but only in the mucinous type of cystadenocarcinoma among ovarian cancer tissue specimens, and cytokeratin was highly expressed in the tissues with a high concentration of CS, which were classified as well-differentiated on the basis of morphological examination. Similarly, cells derived from a keratinizing type of well differentiated cervical carcinoma demonstrated strong potential for synthesis of CS, stained positive with anti-cytokeratin antibody, and exhibited a higher specific activity of TG-1, whereas the cells without CS did not stain positive with anti-cytokeratin antibody and exhibited a lower specific activity of TG-1. These findings indicate that CS is coexpressed with TG-1 and cytokeratin in the well-differentiated types of squamous cell cancers as a tumor marker. PMID- 9865911 TI - Expression of apoptosis-regulating proteins and outcome of esophageal cancer patients treated by combined therapy modalities. AB - The present study retrospectively examines the correlation between the outcome of patients with locally advanced esophageal squamous cell carcinoma (LAEC) after multimodal treatment (radiochemotherapy +/- surgery) and the expression of apoptosis-regulating proteins in pretherapeutic biopsies. Thirty-eight patients with LAEC who took part in a prospective multicentric trial received radiochemotherapy, optionally followed by surgery. Pretreatment tumor biopsies were immunohistochemically investigated for expression of p53, Bcl-2, Bax (bcl-2 associated X protein), and Bcl-X(L) (bcl-2-related X protein). The overall expression of p53, Bcl-2, Bax, and Bcl-X(L) was 52.6, 57.9, 100, and 97.4% respectively. Tumors without p53 expression and tumors with weak Bcl-X(L) expression showed response to chemotherapy more frequently (55.6 and 52.6%, respectively) than tumors positive for p53 expression and tumors with strong Bcl X(L) expression (30.0 and 31.6%, respectively); however, these differences did not attain statistical significance. No correlations were found between the expression of Bcl-2 and Bax and the response to chemotherapy. In patients treated by radiochemotherapy and surgery, p53-negative tumors showed a significantly better outcome than p53-positive tumors (mean survival, 31.1 months versus 11.3 months; P = 0.0378). Additionally, a more favorable outcome was observed in tumors positive for Bcl-2 (not significant), whereas no differences in survival were observed in relation to the expression of Bax or Bcl-X(L). No differences in survival were observed in patients treated by radiochemotherapy without subsequent resection therapy in relation to the expression of apoptosis regulating proteins. Immunohistochemical examination of pretherapeutic tumor biopsies for expression of apoptosis-regulating proteins may help to identify patients with LAEC who may benefit from multimodal treatment and those who may not. PMID- 9865912 TI - Identification of novel mutations in the dihydropyrimidine dehydrogenase gene in a Japanese patient with 5-fluorouracil toxicity. AB - 5-Fluorouracil (5-FU) is used widely in the treatment of several common neoplasms. Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme in the catabolism of 5-FU. Several recent studies have described a pharmacogenetic disorder in which cancer patients with decreased DPD activity develop life-threatening toxicity following exposure to 5-FU. We reported recently the first Japanese case of decreased DPD activity accompanied by severe 5-FU toxicity. The present study describes the results of molecular analysis of this patient and her family, in which three novel mutations (Arg21Gln, Val335Leu, and Glu386Ter) of the gene coding for DPD were identified. We also revealed that Arg21Gln and Glu386Ter are on the same allele and that Val335Leu is on the other allele, on the basis of analysis of the family genome. Expression analysis in Escherichia coli showed that Val335Leu and Glu386Ter led to mutant DPD protein with significant loss of enzymatic activity and no activity, respectively. The Arg21Gln mutation, however, resulted in no decrease in enzymatic activity compared with the wild type. The present data represent the first molecular genetic analysis of DPD deficiency accompanied by severe 5-FU toxicity in a Japanese patient. PMID- 9865913 TI - PTEN mutation in endometrial cancers is associated with favorable clinical and pathologic characteristics. AB - Mutation of the PTEN tumor suppressor gene is a frequent event in endometrial cancers. In other types of cancers, PTEN mutation has been associated with metastatic behavior and advanced stage. To examine the relationship between PTEN mutation and clinical features of endometrial cancers, we screened 136 cases for mutations in the nine exons and intronic splice sites of the PTEN gene, using single-strand conformation analysis, and aberrant bands were sequenced. Mutations were noted in 44 of 136 (32%) endometrial cancers, and two mutations were present in 8 cases. There were 36 cases with mutations resulting in truncated protein products, 6 cases with missense mutations in the phosphatase domain, 1 case with an in-frame deletion, and 1 case with a large insertion. Mutation of the PTEN gene correlated most closely with endometrioid histology; mutations were seen in only 5% (1 of 21) of serous/clear cell cancers compared with 37% (43 of 115) of endometrioid cancers (P = 0.004). PTEN mutation was associated with early stage, nonmetastatic disease and more favorable survival in both the entire group of 136 cases and in the 115 endometrioid cases. In addition, PTEN mutation correlated with other molecular features associated with favorable clinical behavior, including microsatellite instability and absence of p53 overexpression. Microsatellite instability was found in 60% of cases with PTEN mutations compared with only 25% of cases without mutations (P = 0.004). Overexpression of p53 was seen in only 14% of cases with PTEN mutations compared to 39% of cases without mutations (P = 0.006). In conclusion, PTEN mutation is associated with endometrioid histology and other favorable pathological, clinical, and molecular features rather than with increased metastatic potential as has been noted in some other types of cancers. PMID- 9865914 TI - Matrix metalloproteinase-9 expression in bladder washes from bladder cancer patients predicts pathological stage and grade. AB - The matrix metalloproteinases (MMPs), in particular the gelatinases (MMP-2 and MMP-9) have been associated with tumor cell invasion and metastasis in many human cancers. Here we examined the expression of proMMP-2 (gelatinase A) and proMMP-9 (gelatinase B) proteins in the cellular component of bladder washes obtained from 65 patients. Twenty-six patients had active bladder cancer, 24 had a history of bladder cancer but no evidence of active disease at the time of cystoscopy (recurrence-free), and 15 patients had lesions other than bladder cancer (controls). The results were correlated with the cytological findings of the bladder wash and the histopathological results of the tumor resection when performed. In patients with active transitional cell carcinoma of the bladder, 71 and 38% had expression and overexpression of the latent form of MMP-9 (proMMP-9), respectively. In contrast, neither latent nor active MMP-2 could be detected in any of the samples examined, regardless of tumor status. Overexpression of proMMP 9 correlated with higher grade (P = 0.003) and pathological stage (P = 0.04) of disease in the active bladder cancer group. No significant gelatinase expression was detected in the recurrence-free and control cases. Compared with urine cytology, proMMP-9 expression had an overall higher sensitivity for bladder cancer identification (71 versus 54%, P = 0.11). Detection of proMMP-9 in bladder washes may be a novel approach for the identification of patients with more aggressive forms of bladder cancer. PMID- 9865915 TI - Vascular endothelial growth factor, wild-type p53, and angiogenesis in early operable non-small cell lung cancer. AB - Vascular endothelial growth factor (VEGF) is a cytokine that is involved in tumor angiogenesis. Wild-type p53 (wt-p53) protein has been shown in cell lines to suppress angiogenesis through thrombospondin regulation. In this study, we immunohistochemically examined the expression of VEGF, nuclear and wild-type cytoplasmic p53, bcl-2, epidermal growth factor receptor, and c-erbB-2 oncoprotein; vascular grade; proliferation index; and extent of necrosis in non small cell lung cancer (NSCLC). We analyzed 120 cases of early-stage NSCLCs (81 squamous cell carcinomas and 39 adenocarcinomas) treated with surgery alone (median follow-up, 63 months; range, 45-74 months). VEGF expression showed a positive association with high vascular grade (microvessel score of >75 per x250 field; P = 0.008), although about half of the LVG cases also expressed VEGF. None of the p53 antibodies examined correlated with angiogenesis. However, wt-p53 expression was inversely associated with VEGF expression, suggesting that wt-p53 is involved in the suppression of the VEGF gene. Combined analysis of VEGF, wt p53, and microvessel counting showed that, although wt-p53 loss associates with VEGF switch-on, p53 protein may not be involved in the regulation of the angiogenic events downstream of VEGF expression. Moreover, no significant association of bcl-2 and c-erbB-2 oncoprotein expression with VEGF expression was observed. T/N stage, grade, Ki67 proliferation index, and extent of necrosis were not correlated with VEGF expression. Survival analysis showed that VEGF correlated with poor survival (P = 0.04) and was significant in node-negative cases (P = 0.03). We conclude that VEGF is an important angiogenic factor in NSCLC, its expression being dependent on wt-p53 loss. PMID- 9865916 TI - Presence of serum anti-p53 antibodies is associated with pleural effusion and poor prognosis in lung cancer patients. AB - This study was designed prospectively to evaluate the development of anti-p53 antibodies (Abs) in lung cancer patients in relation to their clinical outcome. Sera, derived from 125 lung cancer patients, consisting of 14 small cell lung cancers (SCLC) and 111 non-SCLCs (NSCLC), were surveyed. The p53-null human NSCLC cell line, NCI-H1299, transfected with a human mutant p53 gene was prepared as the source of p53 antigen for immunoblotting analyses to detect the presence of serum anti-p53 Abs. The control group included sera from 10 healthy adults and 14 patients with benign pulmonary diseases. Clinical data including staging and survival were recorded for statistical analyses. The anti-p53 Abs were found in 8% (10 of 125) of the lung cancer patients studied (8.1% of NSCLC versus 7.1% of SCLC patients), whereas none of the control sera had detectable anti-p53 Abs. The presence of anti-p53 Abs was closely associated with malignant pleural effusions (P = 0.001). The p53 Ab-positive patients had a worse prognosis than the p53 Ab negative patients (P < 0.02; median survival, 20 versus 41 weeks). In both univariate and multivariate analyses, the tumor extension and probably the presence of anti-p53 Abs were significant predictors for cancer death. The development of anti-p53 Abs (n = 9) was also a predictor for poor survival in patients with malignant effusions (n = 51). In conclusion, the presence of serum anti-p53 Abs is closely associated with malignant pleural effusions in lung cancer patients. It may serve as a negative prognostic factor for survival independent of malignant pleural effusions and tumor staging. PMID- 9865917 TI - Gamma-ray mutagen sensitivity and survival in patients with glioma. AB - Despite advances in treatment of brain tumors, cerebral malignant gliomas are rapidly debilitating with poor survival. Patient age and tumor histology are known to influence survival in glioma patients, but these factors do not account for all of the variability in survival time. To identify additional useful predictors, we tested an assay that measures intrinsic gamma-ray mutagen sensitivity. Our hypothesis was that increased sensitivity of peripheral blood lymphocytes to chromatid breaks is associated with tumor aggressiveness and decreased patient survival. The eligible 76 patients were those with histologically confirmed malignant gliomas, seen at the University of Texas M. D. Anderson Cancer Center between 1994 and 1997, for whom we had sufficient blood for the in vitro gamma-radiation assay. After gamma-irradiation of each subject's lymphocytes, the frank chromatid breaks in 50 metaphases were averaged to calculate breaks/cell. On the basis of our patient series, we established a gamma ray mutagen sensitivity cutoff point of 0.55 breaks/cell that was confirmed by bootstrap resampling techniques. Patients with values >0.55 breaks/cell were considered sensitive. Kaplan-Meier and Cox proportional hazards modeling were used for the analysis. The gamma-ray mutagen-sensitive patients had worse survival than the nonsensitive patients, with an unadjusted hazard rate ratio of 1.6 (95% confidence interval, 0.9-2.8; P = 0.15). After adjustment for age, tumor histology, and extent of surgical resection, the hazard rate ratio was 2.4 (95% confidence interval, 1.3-4.6; P = 0.0081). Our data suggest that gamma-ray mutagen sensitivity is a prognostic indicator of survival in glioma patients. The significance of these findings needs to be verified in studies with larger samples of patients with histologically similar gliomas. PMID- 9865918 TI - Detection of cancer cells in peripheral blood of breast cancer patients using reverse transcription-polymerase chain reaction for epidermal growth factor receptor. AB - The epidermal growth factor receptor (EGFR) has been reported to be expressed in high levels in primary breast cancer by immunohistochemistry. In the present study, a reverse transcription (RT)-PCR assay using EGFR primers was developed and evaluated for the detection of circulating micrometastases in the blood of breast cancer patients. Total RNA was extracted from breast cancer cell lines and from the blood of 23 control individuals and 37 breast cancer patients. After reverse transcription, outer and nested primers for EGFR were used for cDNA amplification. RNA integrity was confirmed with parallel RT-PCR amplification using beta2-microglobulin primers. PCR products were electrophoresed on agarose gels containing ethidium bromide and visualized by UV photography. Southern blotting was used to confirm EGFR specificity. The nested EGFR RT-PCR assay was capable of detecting a lower limit of 100 fg of total RNA from the A431 cell line. EGFR RNA was identified from the blood of 4 of 18 (22%) metastatic breast cancer patients, 0 of 6 locally recurrent breast cancer patients, 0 of 13 adjuvant breast cancer patients, and 0 of 23 controls (P = 0.03, metastatic versus control). The 18 metastatic breast cancer patients all had progressive disease at the time of blood sampling. The identity of the four EGFR-positive bands was confirmed by Southern blotting. The presence of RT-PCR positivity for EGFR was not a treatment-related phenomenon, because three of the four EGFR positive patients were not receiving treatment at the time of blood collection. RT-PCR for EGFR is a sensitive and specific method for the detection of circulating micrometastases in a proportion of patients with metastatic breast cancer. PMID- 9865919 TI - Profiling of differentially expressed genes in human primary cervical cancer by complementary DNA expression array. AB - The profiling of differentially expressed genes from primary tumor samples using cDNA expression array can reveal new tumor markers as well as target genes for therapeutic intervention. Using cDNA expression array technology, we produced an expression profile of genes that are associated with human cervical cancer. Hybridization of the cDNA blotting membrane (588 genes on a single membrane) was performed with 32P-labeled cDNA probes synthesized from RNA isolated from either normal cervix or cervical cancer. Parallel analyses of the hybridized signals enabled us to profile genes that were differentially expressed in cervical cancer. In each experiment, the extent of hybridization of each gene was evaluated by comparison with the most abundant mRNAs in the human cervix. These include myc proto-oncogene, 40S ribosomal protein S19, heat shock proteins, leukosialin S (CD43), integrin alphaL (CD11A), calgranulin (A), and CDK4 inhibitor (p16ink4). No detectable changes were observed in the expression levels of these genes. Several mRNAs, such as those encoding guanine nucleotide-binding protein Gs (alpha subunit), leukocyte adhesion protein (LFA1-beta), nuclear factor NF45, homeobox protein Hox-A1, and beta-catenin were detected in increased levels in cervical cancer. Genes that showed decreased expression in cervical cancer tissue were a group of apoptosis-related proteins, cell adhesion molecules, nuclear transcription factors, and a homeobox protein (Hox7). For example, the expression levels of Smad1 and Hox7 were consistently decreased in all tumor tissues tested. Northern analysis of Smad1 and Hox7 RNA in primary cervical tumor tissues and cervical carcinoma cell lines indicated that, in general, the mRNA levels of these genes were decreased in human cervical cancer. The precise relationship between the altered expression of these genes and cervical tumorigenesis is a matter of further investigation. PMID- 9865920 TI - The t(8;21) translocation is not consistently associated with high Bcl-2 expression in de novo acute myeloid leukemias of adults. AB - Chromosomal translocations are commonly found in de novo acute myeloid leukemia (AML) cells, and the fusion proteins produced from these genetic abnormalities are assumed to contribute directly to leukemogenesis and/or progression. The AML1/ETO fusion protein, created by translocations between chromosomes 8 and 21 [t(8;21); G. Nucifora and J. D. Rowley, Leuk. Lymphoma, 14: 353-362, 1994; K. L. Rhoades et al., Proc. Natl. Acad. Sci. USA, 93: 11895-11900, 1996] can induce anti-apoptotic Bcl-2 expression in vitro and was proposed to thereby promote the survival of t(8;21)-bearing AML cells (L. Klampfer et al., Proc. Natl. Acad. Sci. USA, 93: 14059-14064, 1996). We confirm that cells of the t(8;21)-bearing Kasumi cell line do express high levels of Bcl-2 protein, as reported previously. However, we show that primary AML cells with (8;21) chromosomal translocations generally express low levels of Bcl-2 protein relative to normal bone marrow derived myeloid cells and to AML samples with other simple karyotypic abnormalities. We note that p53 mutations are present in the myeloid cell lines expressing AML-ETO protein from chromosomal translocations (Kasumi and SKNO) or from transfected fusion genes (U937) but were undetected in our analyses of 28 primary t(8;21)-bearing AML cell samples from de novo AMLs. Because wild-type p53 can transcriptionally down-regulate bcl-2, we speculate that p53 mutations may contribute to the association of t(8;21) chromosomal abnormalities with higher Bcl-2 expression levels in leukemia cell lines. We also note that some t(8;21) bearing samples from pediatric and older adult patients do express somewhat higher levels of Bcl-2 than t(8;21)-bearing samples from young adult patients. This suggests that Bcl-2 overexpression could occur in these AML cells by an as yet undefined, p53-independent mechanism and could contribute to the reported association of t(8;21) karyotypes with poor clinical outcomes in childhood AML patients and/or to typically poor clinical outcomes in elderly AML patients. PMID- 9865921 TI - CDK4 down-regulation induced by paclitaxel is associated with G1 arrest in gastric cancer cells. AB - Paclitaxel induces a cell cycle block at G2-M phase by preventing the depolymerization of microtubules and induces p53-independent apoptosis in many cancer cells. We observed that gastric cancer cells treated with paclitaxel have shown a cyclin-dependent kinase (CDK)4 down-regulation. This paclitaxel-induced CDK4 down-regulation resulted in a cell cycle arrest at G1-S phase. To confirm this observation, we prepared stable transfectants that overexpressed CDK4 and analyzed the cell cycle progression. Ectopic expression of CDK4 in SNU cells resulted in a release of paclitaxel-induced G1 arrest. The release of G1 arrest by enforced expression of CDK4 seems to make the cells more sensitive to paclitaxel-induced apoptosis. From this finding, we could then suggest that paclitaxel treatment induces both G1-S and G2-M blocks in the cell cycle progression of gastric cancer cells. PMID- 9865922 TI - Growth inhibitory effects of aromatic fatty acids on ovarian tumor cell lines. AB - Epithelial ovarian cancer is a major cause of cancer-related mortality in women, making the search for new treatment modalities essential. Sodium phenylacetate (NaPa), a phenylalanine derivative, has been shown to induce cytostasis and differentiation by inhibiting protein isoprenylation. Similar effects have been observed with phenylbutyrate, a phenylacetate congener. We examined in parallel the growth inhibitory activity against human ovarian carcinoma cell lines of phenylacetate, phenylbutyric acid (PB), and certain related compounds, and comparisons were made with lovastatin. On a molar basis, hydroxykynurenine and kynurenine showed the highest activity followed by PB and NaPa. Ovarian carcinoma cell lines were also sensitive to lovastatin in micromolar concentrations. Additive effects were observed when PB was combined with cisplatin or when NaPa or PB were combined with lovastatin. NaPa and PB, but not kynurenine, inhibited incorporation of [3H]mevalonate into ovarian carcinoma cells. An immune modulatory role might also be suggested for PB because it resulted in increased ovarian tumor cell expression of human leukocyte antigen class I and the cluster of differentiation molecule CD58, whereas transforming growth factor-beta2 expression was decreased. Phenylbutyrate, which is the ester form of PB, has shown acceptable pharmacological properties and clinical responses in patients with other malignancies, and might be considered for evaluation in ovarian cancer. PMID- 9865923 TI - Encapsulation of the topoisomerase I inhibitor GL147211C in pegylated (STEALTH) liposomes: pharmacokinetics and antitumor activity in HT29 colon tumor xenografts. AB - The topoisomerase I inhibitor GL147211C [7-[(4-methylpiperazino)methyl]-10,11 (ethylenedioxy)-(20S)-campto thecin trifluoroacetate], a camptothecin analogue, has significant activity in tumor cell cytotoxicity assays in vitro and antitumor activity in both animal tumor models and human patients. Its toxicity is significant, however, effectively limiting the amount of drug that can be administered and its clinical utility. To determine whether the therapeutic index of GL147211C could be improved, the drug was encapsulated in long-circulating, pegylated (STEALTH) liposomes (SPI-355). The pharmacokinetics and antitumor activity of SPI-355 were compared to those of nonliposomal GL147211C. The plasma pharmacokinetics of SPI-355 in rats were typical of those of other pegylated liposomal formulations, with significantly increased blood circulation time; the dose-corrected area under the curve and Cmax of SPI-355 (10 mg/kg) were 1250- and 35-fold higher, respectively, than those of nonliposomal GL14711C (8.72 mg/kg). The comparative antitumor activity of SPI-355 and nonliposomal GL1472211C was evaluated in nude mice implanted with HT29 colon carcinoma xenografts. SPI-355 was 20-fold more effective than GL147211C in inhibiting tumor growth (1 mg/kg SPI 355 and 20 mg/kg GL147211C) and produced durable complete remissions of tumors at well-tolerated dose levels that were >5-fold lower than the maximally tolerated dose of GL147211C, which induced no durable complete responses. Signs of toxicity were similar between the two drugs, but liposome encapsulation increased the toxicity of drug approximately 4-fold, with increased weight loss and several deaths with SPI-355 (5 mg/kg SPI-355 versus 20 mg/kg GL147211C). Despite the increased toxicity seen with SPI-355, the therapeutic index of the liposomal formulation was increased approximately 5-fold over that of nonliposomal GL147211C, suggesting that such a pegylated liposomal formulation could demonstrate increased therapeutic index in human patients. PMID- 9865924 TI - A new screening system for NAD(P)H:quinone oxidoreductase (NQO1)-directed antitumor quinones: identification of a new aziridinylbenzoquinone, RH1, as a NQO1-directed antitumor agent. AB - NAD(P)H:quinone oxidoreductase (NQO1; DT-diaphorase) is elevated in certain tumors, such as non-small cell lung cancer (NSCLC). Compounds such as mitomycin C and streptonigrin are efficiently bioactivated by NQO1 and have been used in an enzyme-directed approach to chemotherapy. Previously, 2,5-diaziridinyl-3,6 dimethyl-1,4-benzoquinone (MeDZQ) was identified as a potential antitumor agent based on its high rate of bioactivation by human NQO1 and its selective cytotoxicity to cells containing elevated NQO1. RH1, a water-soluble analogue of MeDZQ synthesized in this work, was a better substrate for recombinant human NQO1 than the parent compound. RH1 was, correspondingly, more cytotoxic to human tumor cells expressing elevated NQO1 activity (H460 NSCLC and HT29 human colon carcinoma), as measured by 3-(4,5-dimethylthiazol-2,5-diphenyl)tetrazolium assay, than it was to cells deficient in NQO1 activity (H596 NSCLC and BE human colon carcinoma). RH1 exhibited a greater selective toxicity (ratio of IC50s in H596:H460 and BE:HT29) to cells with elevated NQO1 activity relative to MeDZQ. Additionally, we report the establishment of a stable line of BE human colon carcinoma cells transfected with wild-type human NQO1 (BE-NQ7). BE cells are devoid of NQO1 activity due to a homozygous point mutation in the NQO1 gene. In comparison to the parental cell line, RH1, MeDZQ, and mitomycin C were significantly more cytotoxic to BE-NQ7 cells (17-, 7-, and 3-fold, respectively), confirming that the presence of NQO1 is sufficient to increase cytotoxicity of these antitumor quinones. These data suggest that RH1 may be an effective NQO1 directed antitumor agent for the therapy of tumors with elevated NQO1 activity, such as NSCLC. PMID- 9865925 TI - Conversion of the CPT-11 metabolite APC to SN-38 by rabbit liver carboxylesterase. AB - The anticancer drug CPT-11 (7-ethyl-[4(1-piperidino)-1 piperidino]carbonyloxycamptothecin) is a water-soluble derivative of camptothecin. We report here the conversion of APC (7-ethyl-[4-N-(5 aminopentanoic acid)-1-piperidino] carbonyloxycamptothecin), an inactive metabolite of CPT-11, to SN-38 (7-ethyl-10-hydroxycamptothecin), the active metabolite of CPT-11, by a rabbit liver carboxylesterase. This reaction is not catalyzed by any known human enzyme. The formation of SN-38 from APC was characterized by an apparent Km of 37.9 +/- 7.1 microM and a Vmax of 16.9 +/- 0.9 pmol/units/min. SN-38 was confirmed as a reaction product by high-performance liquid chromatography and mass spectrometry. A 24-h incubation of 10 microM APC with 500 units/ml of rabbit carboxylesterase produced 4 microM SN-38. The product of this reaction inhibited the growth of U373 MG human glioblastoma cells in vitro. The IC50 for a 24-h exposure of U373 MG cells to APC in the presence of 50 units/ml of rabbit carboxylesterase was 0.27 +/- 0.08 microM, whereas APC alone demonstrated no inhibition of growth at concentrations up to 1 microM. The IC50 of U373 MG cells transfected with the cDNA encoding the rabbit carboxylesterase (U373pIRESrabbit) and exposed to APC for 24 h was 0.8 +/- 0.1 microM APC, whereas the growth of cells transfected with vector control (U373pIRES) was unaffected by up to 1 microM APC. Because APC is nontoxic to human cells, we are investigating the possibility of using APC/rabbit carboxylesterase in a prodrug/enzyme therapeutic approach. PMID- 9865926 TI - Graft-versus-leukemia reactivity involves cluster formation between superantigen reactive donor T lymphocytes and host macrophages. AB - T-cell-mediated antitumor effects play an important role clinically in allogeneic graft-versus-leukemia (GvL) reactivity, whereas T-cell-mediated antihost effects are associated with a risk of developing graft-versus-host (GvH) disease. GvL and GvH were compared in an animal tumor model system after the systemic transfer of allogeneic antitumor immune T lymphocytes from B10.D2 [H-2d; minor lymphocyte stimulating antigen (Mls)b] mice into ESb-MP tumor-bearing or normal DBA/2 (H-2d; Mls(a)) mice. Here we demonstrate that this T-cell-mediated therapy involves the formation of clusters of donor CD4 and CD8 T cells with host macrophages, in particular, with a subpopulation expressing the lymphocyte adhesion molecule sialoadhesin. DBA/2 mice and the derived tumor ESb-MP express viral superantigen 7 (Mls(a)), an endogenous viral superantigen that is absent from B10.D2 mice. To test the contribution of viral superantigen 7-reactive Vbeta6 donor T cells in the GvL-mediated eradication of liver metastases, we performed immunohistological and transmission electron microscopy studies. Vbeta6+ CD4 and CD8 T cells from B10.D2 donors formed tight clusters with host sialoadhesin-positive macrophages, and transmission electron microscopy pictures revealed direct membrane-membrane interactions between T cells and macrophages. Clusters were more abundant and consisted of more cells in tumor-bearing hosts (GvL model) than in non-tumor bearing hosts (GvH model). In addition, Vbeta6 T cells within the clusters showed a strong proliferation activity, indicating stimulation. Moreover, in an in vitro tumor cytostasis assay, primed as well as nonprimed purified Vbeta6 T cells from donor mice were able to inhibit the proliferation of superantigen-expressing ESb MP lymphoma cells. This suggests that the transferred superantigen-reactive Vbeta6 T cells contribute to the eradication of metastases. The observed cell clusters might be sites for antigen presentation and the activation of tumor reactive T cells. PMID- 9865927 TI - Transplantation of fetal brain tissue into the site of intracerebral hemorrhage in rats. AB - The purpose of this study was to evaluate whether transplantation of fetal forebrain tissue into the hematoma site of rats with intrastriatal hemorrhage could improve the final neurological outcome. Nine to twelve days after collagenase-induced intracerebral hemorrhage, day 14 fetal forebrain fragments were transplanted into hematoma site. Quantitative measures of behavioral function were repeatedly evaluated until the rats were killed 10 weeks after grafting. Histology was used to assess the survival of the grafts and overall brain morphology. Surviving grafts were located in the residual cavity at the hematoma site. However, comparison of rats with live transplants to control rats with no transplant, sham transplant, or dead tissue transplant revealed no statistically significant differences in any of the motor tests. PMID- 9865928 TI - Reduced tyrosine kinase receptor C mRNA levels in the frontal cortex of patients with schizophrenia. AB - Using a quantitative RNA-PCR approach tyrosine kinase receptor (trk) C mRNA levels were determined in brain material from the frontal cortex (BA10), temporal cortex (BA20) and cerebellum of control specimen and patients with schizophrenia, bipolar disorder or non-psychotic depression (15 subjects each). In the frontal cortex of schizophrenics there was a 5.8-fold reduction of trk C mRNA levels, which reached statistical significance (P < 0.05). Trk C levels in the cerebellum were positively correlated with lifetime fluphenazine equivalents (r = 0.54), suggesting that neuroleptics influence TRK C gene activity in the cerebellum. Moreover, the distinct medication-independent reduction of trk C mRNA may point to a disturbed neurotrophic gene activity in the frontal cortex of schizophrenic patients. PMID- 9865929 TI - Temporal integration in a subjective accentuation task as a function of child cognitive development. AB - We have previously shown that temporal integration in the domain of a few seconds may be studied using a subjective accentuation paradigm. Here we report developmental effects on the limits of this temporal integration in 9-10-year olds in comparison with 13-14-year-olds. The task was to listen to a string of identical metronome beats and mentally bind the presented beats by subjectively accentuating every second, third or nth beat. The integration interval length was defined as the number of stimuli mentally connected multiplied by the temporal interval between two successive beats. For the lowest stimulus frequency integration intervals were approximately 3 s for the older and 2.2 s for the younger children. For higher frequencies integration intervals got systematically shorter, but being always longer for the older age group. It is suggested that the prefrontal region is responsible for this developmental effect. The expansion of temporal integration correlates with cognitive development in the investigated phase of ontogenesis. PMID- 9865930 TI - Prediction of migraine attacks using a slow cortical potential, the contingent negative variation. AB - Amplitudes of contingent negative variation (CNV) as a slow cortical potential in migraine patients were recorded before and after migraine attacks. The level of CNV amplitudes, especially of the early component, showed a trend in rise of negativity 1 day before the attack, whereas amplitudes were normal for 2-3 days following an attack. In most cases the attack occurred when the CNV amplitude showed maximum negativity. We conclude that migraine-susceptible days are indicated by a high CNV amplitude the day before the attack. PMID- 9865931 TI - Soluble Alzheimers beta-amyloid constricts the cerebral vasculature in vivo. AB - Bilateral temporoparietal hypoperfusion has been frequently observed early in the Alzheimer's disease (AD) process. An increased beta-amyloid (Abeta) peptide is believed to play a central role in the pathogenesis of AD. In vitro experiments have shown that freshly solubilized Abeta enhances constriction of cerebral and peripheral vessels. We propose that in vivo the Abeta vasoactive property may contribute to cerebral hypoperfusion of AD patients. To test this hypothesis, we intra-arterially infused freshly solubilized Abeta -40 in rats and observed changes in cerebral blood flow and cerebrovascular resistance using fluorescent microspheres. We found that infusion of Abeta in vivo resulted in a decreased blood flow and increased vascular resistance specifically in cerebral cortex but not in heart or kidneys. These data suggest that Abeta has a direct and specific constrictive effect on cerebral vessels in vivo, which may contribute to the cerebral hypoperfusion observed early in the AD process. PMID- 9865932 TI - Excitotoxic effect of kainic acid on chicken cochlear afferent neurons. AB - The excitotoxic effects of kainic acid, a glutamate analog, on the auditory neurons in the chicken cochlea were assessed by light and transmission electron microscopy. Kainic acid was directly applied onto the round window of adult chickens and their cochleas were harvested 3 h after application. Transverse microscopic sections of the basilar papilla revealed swelling of afferent dendrites without any morphological changes in efferent endings. The regions of the basilar papilla damaged by kainic acid were localized in the apical 80% and primarily on the neural side where tall hair cells are located. The basal, abneural short hair cell region was devoid of damage. These results imply that glutamate is a primary neurotransmitter in chicken auditory afferent neurons that synapse on tall hair cells. PMID- 9865933 TI - Brain-derived neurotrophic factor does not prevent ionizing radiation-induced apoptosis in the developing rat brain. AB - In an attempt to learn whether brain-derived neurotrophic factor (BDNF) is able to protect neural cells from ionizing radiation-induced apoptosis in the developing brain, BDNF-transfected (F3N-BDNF) and mock-transfected (F3A MT) fibroblasts were grafted into the right germinal periventricular zone of rats at postnatal day 1 (P1). The rats were later irradiated at P3 with a single dose (2 Gy whole body irradiation) of gamma rays. Age-matched non-irradiated and non grafted irradiated rats at P3 were used as controls. No differences in the number of apoptotic cells were observed between the ipsilateral and contralateral side of either F3N-BDNF- or F3A-MT-grafted irradiated rats Furthermore, counts of apoptotic cells were similar in non-grafted irradiated and grafted irradiated rats. These results show that BDNF has no capacity to prevent ionizing radiation induced apoptosis in the developing rat brain. PMID- 9865934 TI - Immunosuppressive effect of vestibulo-cerebellar lesion in rats. AB - Kainate lesion of the vestibulo cerebellum induces sympathetic hyperactivity, but the mechanism of immunosuppression observed as a result is not yet clarified. Here we report that vestibulo cerebellum lesioned (VCL) rats have depressed secretion of haematopoietic cytokines (bioimmunomodulator or BIM, a 12.7 kD peptide and thymosin FrV) in tissue cultures of bone marrow and thymus, respectively, compared with controls (P < 0.01). Peripheral blood leukocyte concentration, neutrophil myeloperoxydase response, T-SRBC rosette and antibody titre to sheep red blood cells (SRBC) are also significantly less, compared with control (P < 0.01). Injection of BIM (concentration 0.01 microg/g body weight) in VCL rats corrected the immunodeficiency. Partial restoration of immune competence is observed after injection of thymosin FrV (0.01 microg/g body weight) or after prolonged vestibular stimulation (18 rpm for 15 min/day for 21 days). The results indicate that the vestibular nodule (VN) through autonomic nerves (AN) can modulate the immune function of rats by regulating the secretion of cytokines from bone marrow and thymus. PMID- 9865935 TI - Phospholipases as mediators of amyloid beta peptide neurotoxicity: an early event contributing to neurodegeneration characteristic of Alzheimer's disease. AB - There is a consensus that by some still to be defined mechanism amyloid beta peptide, which accumulates in Alzheimer's disease brain tissue, contributes to the characteristic neurodegeneration. We suggest that one of these mechanisms for amyloid beta peptide is the ability to activate cellular phospholipases. Excessive phospholipid hydrolysis would produce a variety of lipidic second messengers. These catabolites would then evoke unnecessary stereotypic responses. This indiscriminate activation of the phospholipases could be responsible for the increased amounts of phospholipid catabolites found in Alzheimer's disease brain tissue. Failure to maintain regeneration of the membrane components would result in a loss of essential cellular neuronal processes. PMID- 9865936 TI - Localization of a novel inositol 1,4,5-trisphosphate binding protein, p130 in rat brain. AB - We have isolated a novel inositol 1,4,5-trisphosphate binding protein with molecular mass of 130 kDa (p130), homologous to phospholipase C-delta1 in amino acid sequence but with no catalytic activity. Here we report the expression and localization of p130 at the mRNA level in rat brain. Northern blotting showed that gene expression encoding p130 was most abundant in brain. Brain localization of p130-mRNA using an in situ hybridization technique revealed that in the cerebellum, the mRNA was detected in the granular cell and Purkinje cell layers, and cerebellar nuclei. In the cerebrum, the mRNA was localized in hippocampal pyramidal cells, dentate granule cells and pyramidal and/or granule cells of the cerebral cortex. The brain localization of p130-mRNA was similar to that of the beta-subtype of phospholipase C, indicating that p130 may be mainly involved in phospholipase Cbeta-mediated signaling. PMID- 9865937 TI - Frequency and synchrony of tetanically-induced, gamma-frequency population discharges in the rat hippocampal slice: the effect of diazepam and propofol. AB - Fast, rhythmic discharges, at gamma frequencies, have been evoked in the CA1 region of the hippocampus by tetanic stimulation applied via pairs of electrodes placed close to the pyramidal layer. The discharges had a latency of 65-225 ms and a frequency of 60-100 Hz; they were synchronous (phase lag < 2 ms) when recorded from sites 0.4-0.8 mm apart. Diazepam (5-15 microM) and propofol (5 microM) reduced the frequency by 15-26%, but had no significant effect on synchrony or phase lag, suggesting that the generation and synchrony of this rhythm are pharmacologically distinct. 10 microM propofol had a more marked effect and, when frequency fell by approximately 50%, synchrony was also reduced. PMID- 9865938 TI - Temporal profile of magnetic resonance imaging changes following forebrain ischemia in the gerbil. AB - Quantitative T2 magnetic resonance (MR) imaging was used to examine gerbil brains 1, 3, 10, and 30 days after 5 min forebrain ischemia. T2 was increased in the dorsal-lateral striatum 1 and 3 days post-ischemia, and in the hippocampus 3 days post-ischemia. T2 was normal 10 days post-ischemia, and decreased in the hippocampus and dorsal-lateral striatum 30 days post-ischemia. Neuronal counts in the dorsal-lateral striatum and CA1 hippocampal region were uniformly decreased 30 days post-ischemia. The increase in T2 shortly after ischemia is attributed to brain edema localized to regions where neuronal injury developed. The late decrease in T2 may be due to decreased water in gliotic tissue, or to ferritin positive microglia, following forebrain ischemia. Tissue atrophy at later times gave enlarged ventricles on MR images. PMID- 9865939 TI - Mechanisms of fatigue-induced activation of group IV muscle afferents: the roles played by lactic acid and inflammatory mediators. AB - We already showed that group IV muscle afferents are activated during electrically-induced fatigue. The purpose of this study is to identify the mechanisms of stimulation of these muscle afferents by electrically-induced fatigue at a high (100 Hz; high frequency fatigue, HFF) or a low rate (10 Hz; low frequency fatigue, LFF) of stimulation. In 23 paralyzed and anaesthetized rabbits, group IV afferent activity from the tibialis anterior muscle was recorded before and after 3-min HFF or 5-min LFF runs eliciting the same force failure. Plasma lactic acid concentration (LA) was also measured in leg venous blood. We tested the effects of dichloroacetate (DCA), which reduces lactic acid production, and of acetylsalicylic acid (ASA), a blocker of cyclooxygenase, on fatigue-induced activation of group IV muscle afferents after HFF and LFF trials. Pretreatment by ASA or DCA did not modify HFF-induced activation of muscle afferents. On the other hand, LFF-induced response was markedly depressed by each pharmacological agent (-44% after ASA and -75% after DCA). We verified that DCA markedly lowered LA production after LFF. The present results show that the activation of group IV muscle afferents by LFF or HFF results from different mechanisms and also demonstrate the major role played by lactic acid production and, to a lesser extent, the release of inflammatory mediators in LFF-induced activation of group IV muscle afferents. PMID- 9865940 TI - The expression of late infantile neuronal ceroid lipofuscinosis (CLN2) gene product in human brains. AB - We raised polyclonal antibodies against a gene product responsible for late infantile neuronal ceroid lipofuscinosis (CLN2). By Western blotting, all three antisera recognized the CLN2 protein at approximately 49 kDa in human brain homogenates. Immunohistochemistry using the antisera demonstrated the granular labelling in the cytoplasm of cerebral neurons and glial cells. The immunoreactivity on Western blots was absent from the brain of a patient with CLN2. Our results suggest the usefulness of these antibodies for the diagnosis of CLN2, which currently requires demonstration of characteristic ultrastructure by electron microscopy. PMID- 9865941 TI - Antiphospholipid antibodies bind to rat cerebellar granule cells: the role of N methyl-D-aspartate receptors. AB - IgGs from sera containing antiphospholipid antibodies (aPL), detected as antibodies to cardiolipin, or control sera were incubated with rat cerebellar granule cells in primary culture. Using a mitochondrial dehydrogenase activity assay (MTT test), aPL IgGs were shown to decrease MTT metabolism after 24 h incubation with the cells, and to cause non-toxic amounts of glutamate to become neurotoxic when added to the cells for 45 min. Acute and chronic aPL toxicity were prevented by MK-801. Sera containing aPL bound to intact cerebellar neurons, as revealed by an immunofluorescent technique. These results suggest that antiphospholipid antibodies interfere with excitatory pathways in glutamatergic cerebellar granule cells by a mechanism involving overactivation of the NMDA glutamate receptor. PMID- 9865942 TI - Structural analysis of inhibitor binding to human carbonic anhydrase II. AB - X-ray crystal structures of carbonic anhydrase II (CAII) complexed with sulfonamide inhibitors illuminate the structural determinants of high affinity binding in the nanomolar regime. The primary binding interaction is the coordination of a primary sulfonamide group to the active site zinc ion. Secondary interactions fine-tune tight binding in regions of the active site cavity >5 A away from zinc, and this work highlights three such features: (1) advantageous conformational restraints of a bicyclic thienothiazene-6-sulfonamide 1,1-dioxide inhibitor skeleton in comparison with a monocyclic 2,5 thiophenedisulfonamide skeleton; (2) optimal substituents attached to a secondary sulfonamide group targeted to interact with hydrophobic patches defined by Phe131, Leu198, and Pro202; and (3) optimal stereochemistry and configuration at the C-4 position of bicyclic thienothiazene-6-sulfonamides; the C-4 substituent can interact with His64, the catalytic proton shuttle. Structure-activity relationships rationalize affinity trends observed during the development of brinzolamide (Azopt), the newest carbonic anhydrase inhibitor approved for the treatment of glaucoma. PMID- 9865943 TI - Solution structure of barley lipid transfer protein complexed with palmitate. Two different binding modes of palmitate in the homologous maize and barley nonspecific lipid transfer proteins. AB - The structure of a nonspecific lipid transfer protein from barley (ns-LTPbarley) in complex with palmitate has been determined by NMR spectroscopy. The structure has been compared to the structure of ns-LTPbarley in the absence of palmitate, to the structure of ns-LTPbarley in complex with palmitoyl coenzyme A, to the structure of ns-LTPmaize in its free form, and to the maize protein complexed with palmitate. Binding of palmitate only affects the structure of ns-LTPbarley moderately in contrast to the binding of palmitoyl coenzyme A, which leads to a considerable expansion of the protein. The modes of binding palmitate to the maize and barley protein are different. Although in neither case there are major conformational changes in the protein, the orientation of the palmitate in the two proteins is exactly opposite. PMID- 9865944 TI - A homology identification method that combines protein sequence and structure information. AB - A new method is presented for identifying distantly related homologous proteins that are unrecognizable by conventional sequence comparison methods. The method combines information about functionally conserved sequence patterns with information about structure context. This information is encoded in stochastic discrete state-space models (DSMs) that comprise a new family of hidden Markov models. The new models are called sequence-pattern-embedded DSMs (pDSMs). This method can identify distantly related protein family members with a high sensitivity and specificity. The method is illustrated with trypsin-like serine proteases and globins. The strategy for building pDSMs is presented. The method has been validated using carefully constructed positive and negative control sets. In addition to the ability to recognize remote homologs, pDSM sequence analysis predicts secondary structures with higher sensitivity, specificity, and Q3 accuracy than DSM analysis, which omits information about conserved sequence patterns. The identification of trypsin-like serine proteases in new genomes is discussed. PMID- 9865945 TI - Crystal structure of the effector-binding domain of the trehalose-repressor of Escherichia coli, a member of the LacI family, in its complexes with inducer trehalose-6-phosphate and noninducer trehalose. AB - The crystal structure of the Escherichia coli trehalose repressor (TreR) in a complex with its inducer trehalose-6-phosphate was determined by the method of multiple isomorphous replacement (MIR) at 2.5 A resolution, followed by the structure determination of TreR in a complex with its noninducer trehalose at 3.1 A resolution. The model consists of residues 61 to 315 comprising the effector binding domain, which forms a dimer as in other members of the LacI family. This domain is composed of two similar subdomains each consisting of a central beta sheet sandwiched between alpha-helices. The effector binding pocket is at the interface of these subdomains. In spite of different physiological functions, the crystal structures of the two complexes of TreR turned out to be virtually identical to each other with the conformation being similar to those of the effector binding domains of the LacI and PurR in complex with their effector molecules. According to the crystal structure, the noninducer trehalose binds to a similar site as the trehalose portion of trehalose-6-phosphate. The binding affinity for the former is lower than for the latter. The noninducer trehalose thus binds competitively to the repressor. Unlike the phosphorylated inducer molecule, it is incapable of blocking the binding of the repressor headpiece to its operator DNA. The ratio of the concentrations of trehalose-6-phosphate and trehalose thus is used to switch between the two alternative metabolic uses of trehalose as an osmoprotectant and as a carbon source. PMID- 9865946 TI - Identification of kinetically hot residues in proteins. AB - A number of recent studies called attention to the presence of kinetically important residues underlying the formation and stabilization of folding nuclei in proteins, and to the possible existence of a correlation between conserved residues and those participating in the folding nuclei. Here, we use the Gaussian network model (GNM), which recently proved useful in describing the dynamic characteristics of proteins for identifying the kinetically hot residues in folded structures. These are the residues involved in the highest frequency fluctuations near the native state coordinates. Their high frequency is a manifestation of the steepness of the energy landscape near their native state positions. The theory is applied to a series of proteins whose kinetically important residues have been extensively explored: chymotrypsin inhibitor 2, cytochrome c, and related C2 proteins. Most of the residues previously pointed out to underlie the folding process of these proteins, and to be critically important for the stabilization of the tertiary fold, are correctly identified, indicating a correlation between the kinetic hot spots and the early forming structural elements in proteins. Additionally, a strong correlation between kinetically hot residues and loci of conserved residues is observed. Finally, residues that may be important for the stability of the tertiary structure of CheY are proposed. PMID- 9865947 TI - Pulmonary surfactant-associated polypeptide C in a mixed organic solvent transforms from a monomeric alpha-helical state into insoluble beta-sheet aggregates. AB - In the 35-residue pulmonary surfactant-associated lipopolypeptide C (SP-C), the stability of the valyl-rich alpha-helix comprising residues 9-34 has been monitored by circular dichroism, nuclear magnetic resonance, and Fourier transform infrared spectroscopy in both a mixed organic solvent and in phospholipid micelles. The alpha-helical form of SP-C observed in freshly prepared solutions in a mixed solvent of CHCl3/CH3OH/0.1 M HCl 32:64:5 (v/v/v) at 10 degrees C undergoes within a few days an irreversible transformation to an insoluble aggregate that contains beta-sheet secondary structure. Hydrogen exchange experiments revealed that this conformational transition proceeds through a transition state with an Eyring free activation enthalpy of about 100 kJ mol(-1), in which the polypeptide segment 9-27 largely retains a helical conformation. In dodecylphosphocholine micelles, the helical form of SP-C was maintained after seven weeks at 50 degrees C. The alpha-helical form of SP-C thus seems to be the thermodynamically most stable state in this micellar environment, whereas its presence in freshly prepared samples in the aforementioned mixed solvent is due to a high kinetic barrier for unfolding. These observations support a previously proposed pathway for in vivo synthesis of SP-C through proteolytic processing from a 21-kDa precursor protein. PMID- 9865948 TI - The crystal structure of NADPH:ferredoxin reductase from Azotobacter vinelandii. AB - NADPH:ferredoxin reductase (AvFPR) is involved in the response to oxidative stress in Azotobacter vinelandii. The crystal structure of AvFPR has been determined at 2.0 A resolution. The polypeptide fold is homologous with six other oxidoreductases whose structures have been solved including Escherichia coli flavodoxin reductase (EcFldR) and spinach, and Anabaena ferredoxin:NADP+ reductases (FNR). AvFPR is overall most homologous to EcFldR. The structure is comprised of a N-terminal six-stranded antiparallel beta-barrel domain, which binds FAD, and a C-terminal five-stranded parallel beta-sheet domain, which binds NADPH/NADP+ and has a classical nucleotide binding fold. The two domains associate to form a deep cleft where the NADPH and FAD binding sites are juxtaposed. The structure displays sequence conserved motifs in the region surrounding the two dinucleotide binding sites, which are characteristic of the homologous enzymes. The folded over conformation of FAD in AvFPR is similar to that in EcFldR due to stacking of Phe255 on the adenine ring of FAD, but it differs from that in the FNR enzymes, which lack a homologous aromatic residue. The structure of AvFPR displays three unique features in the environment of the bound FAD. Two features may affect the rate of reduction of FAD: the absence of an aromatic residue stacked on the isoalloxazine ring in the NADPH binding site; and the interaction of a carbonyl group with N10 of the flavin. Both of these features are due to the substitution of a conserved C-terminal tyrosine residue with alanine (Ala254) in AvFPR. An additional unique feature may affect the interaction of AvFPR with its redox partner ferredoxin I (FdI). This is the extension of the C-terminus by three residues relative to EcFldR and by four residues relative to FNR. The C-terminal residue, Lys258, interacts with the AMP phosphate of FAD. Consequently, both phosphate groups are paired with a basic group due to the simultaneous interaction of the FMN phosphate with Arg51 in a conserved FAD binding motif. The fourth feature, common to homologous oxidoreductases, is a concentration of 10 basic residues on the face of the protein surrounding the active site, in addition to Arg51 and Lys258. PMID- 9865949 TI - Dominant role of local dipolar interactions in phosphate binding to a receptor cleft with an electronegative charge surface: equilibrium, kinetic, and crystallographic studies. AB - Stringent specificity and complementarity between the receptor, a periplasmic phosphate-binding protein (PBP) with a two-domain structure, and the completely buried and dehydrated phosphate are achieved by hydrogen bonding or dipolar interactions. We recently found that the surface charge potential of the cleft between the two domains that contains the anion binding site is intensely electronegative. This novel finding prompted the study reported here of the effect of ionic strength on the equilibrium and rapid kinetics of phosphate binding. To facilitate this study, Ala197, located on the edge of the cleft, was replaced by a Trp residue (A197W PBP) to generate a fluorescence reporter group. The A197W PBP-phosphate complex retains wild-type Kd and X-ray structure beyond the replacement residue. The Kd (0.18 microM) at no salt is increased by 20-fold at greater than 0.30 M NaCl. Stopped-flow fluorescence kinetic studies indicate a two-step binding process: (1) The phosphate (L) binds, at near diffusion controlled rate, to the open cleft form (Po) of PBP to produce an intermediate, PoL. This rate decreases with increasing ionic strength. (2) The intermediate isomerizes to the closed-conformation form, PcL. The results indicate that the high specificity, affinity, and rate of phosphate binding are not influenced by the noncomplementary electronegative surface potential of the cleft. That binding depends almost entirely on local dipolar interactions with the receptor has important ramification in electrostatic interactions in protein structures and in ligand recognition. PMID- 9865950 TI - Crystal structure of two quaternary complexes of dethiobiotin synthetase, enzyme MgADP-AlF3-diaminopelargonic acid and enzyme-MgADP-dethiobiotin-phosphate; implications for catalysis. AB - The crystal structures of two complexes of dethiobiotin synthetase, enzyme diaminopelargonic acid-MgADP-AlF3 and enzyme-dethiobiotin-MgADP-Pi, respectively, have been determined to 1.8 A resolution. In dethiobiotin synthetase, AlF3 together with carbamylated diaminopelargonic acid mimics the phosphorylated reaction intermediate rather than the transition state complex for phosphoryl transfer. Observed differences in the binding of substrate, diaminopelargonic acid, and the product, dethiobiotin, suggest considerable displacements of substrate atoms during the ring closure step of the catalytic reaction. In both complexes, two metal ions are observed at the active site, providing evidence for a two-metal mechanism for this enzyme. PMID- 9865951 TI - Interaction of wheat alpha-thionin with large unilamellar vesicles. AB - The interaction of the wheat antibacterial peptide alpha-thionin with large unilamellar vesicles has been investigated by means of fluorescence spectroscopy. Binding of the peptide to the vesicles is followed by the release of vesicle contents, vesicle aggregation, and lipid mixing. Vesicle fusion, i.e., mixing of the aqueous contents, was not observed. Peptide binding is governed by electrostatic interactions and shows no cooperativity. The amphipatic nature of wheat alpha-thionin seems to destabilize the membrane bilayer and trigger the aggregation of the vesicles and lipid mixing. The presence of distearoylphosphatidylethanolamine-poly(ethylene glycol 2000) (PEG-PE) within the membrane provides a steric barrier that inhibits vesicle aggregation and lipid mixing but does not prevent leakage. Vesicle leakage through discrete membrane channels is unlikely, because the release of encapsulated large fluorescent dextrans is very similar to that of 8-aminonaphthalene-1,3,6,trisulfonic acid (ANTS). A minimum number of 700 peptide molecules must bind to each vesicle to produce complete leakage, which suggests a mechanism in which the overall destabilization of the membrane is due to the formation of transient pores rather than discrete channels. PMID- 9865952 TI - Empirical solvent-mediated potentials hold for both intra-molecular and inter molecular inter-residue interactions. AB - Whether knowledge-based intra-molecular inter-residue potentials are valid to represent inter-molecular interactions taking place at protein-protein interfaces has been questioned in several studies. Differences in the chain connectivity effect and in residue packing geometry between interfaces and single chain monomers have been pointed out as possible sources of distinct energetics for the two cases. In the present study, the interfacial regions of protein-protein complexes are examined to extract inter-molecular inter-residue potentials, using the same statistical methods as those previously adopted for intra-molecular residue pairs. Two sets of energy parameters are derived, corresponding to solvent-mediation and "average residue" mediation. The former set is shown to be highly correlated (correlation coefficient 0.89) with that previously obtained for inter-residue interactions within single chain monomers, while the latter exhibits a weaker correlation (0.69) with its intra-molecular counterpart. In addition to the close similarity of intra- and inter-molecular solvent-mediated potentials, they are shown to be significantly more residue-specific and thereby discriminative compared to the residue-mediated ones, indicating that solvent mediation plays a major role in controlling the effective inter-residue interactions, either at interfaces, or within single monomers. Based on this observation, a reduced set of energy parameters comprising 20 one-body and 3 two body terms is proposed (as opposed to the 20 x 20 tables of inter-residue potentials), which reproduces the conventional 20 x 20 tables with a correlation coefficient of 0.99. PMID- 9865953 TI - Recognition of partially-folded mitochondrial malate dehydrogenase by GroEL. Steady and time-dependent emission anisotropy measurements. AB - The binding of partially-folded mitochondrial malate dehydrogenase (mMDH) to GroEL was assessed by steady and nanosecond emission spectroscopy. Partially folded intermediates of mMDH show significant residual secondary structure when examined by CD spectroscopy in the far UV. They bind the extrinsic fluorescent probe ANS and the protein-ANS complexes display a rotational correlation time of 19 ns. Similar rotational correlation time (phi = 18.6 ns) was determined for partially-folded species tagged with anthraniloyl. GroEL recognizes partially folded species with a K(D) approximately 60 nM. The rotational correlation time of the complex, i.e., GroEL-mMDH-ANT, approaches a value of 280 ns in the absence of ATP. Reactivation of mMDH-ANT by addition of GroEL and ATP brings about a significant decrease in the observed rotational correlation time. The results indicate that partially-folded malate dehydrogenase is rigidly trapped by GroEL in the absence of ATP, whereas addition of ATP facilitates reactivation and release of folded conformations endowed with catalytic activity. PMID- 9865954 TI - Domain structure analysis of elongation factor-3 from Saccharomyces cerevisiae by limited proteolysis and differential scanning calorimetry. AB - Elongation-factor-3 (EF-3) is an essential factor of the fungal protein synthesis machinery. In this communication the structure of EF-3 from Saccharomyces cerevisiae is characterized by differential scanning calorimetry (DSC), ultracentrifugation, and limited tryptic digestion. DSC shows a major transition at a relatively low temperature of 39 degrees C, and a minor transition at 58 degrees C. Ultracentrifugation shows that EF-3 is a monomer; thus, these transitions could not reflect the unfolding or dissociation of a multimeric structure. EF-3 forms small aggregates, however, when incubated at room temperature for an extended period of time. Limited proteolysis of EF-3 with trypsin produced the first cleavage at the N-side of Gln775, generating a 90-kDa N-terminal fragment and a 33-kDa C-terminal fragment. The N-terminal fragment slowly undergoes further digestion generating two major bands, one at approximately 75 kDa and the other at approximately 55 kDa. The latter was unusually resistant to further tryptic digestion. The 33-kDa C-terminal fragment was highly sensitive to tryptic digestion. A 30-min tryptic digest showed that the N-terminal 60% of EF-3 was relatively inaccessible to trypsin, whereas the C terminal 40% was readily digested. These results suggest a tight structure of the N-terminus, which may give rise to the 58 degrees C transition, and a loose structure of the C-terminus, giving rise to the 39 degrees C transition. Three potentially functional domains of the protein were relatively resistant to proteolysis: the supposed S5-homologous domain (Lys102-Ile368), the N-terminal ATP-binding cassette (Gly463-Lys622), and the aminoacyl-tRNA-synthase homologous domain (Glu820-Gly865). Both the basal and ribosome-stimulated ATPase activities were inactivated by trypsin, but the ribosome-stimulated activity was inactivated faster. PMID- 9865955 TI - Localization of basic residues required for receptor binding to the single alpha helix of the receptor binding domain of human alpha2-macroglobulin. AB - To better understand the structural basis for the binding of proteinase transformed human alpha2-macroglobulin (alpha2M) to its receptor, we have used three-dimensional multinuclear NMR spectroscopy to determine the secondary structure of the receptor binding domain (RBD) of human alpha2M. Assignment of the backbone NMR resonances of RBD was made using 13C/15-N and 15N-enriched RBD expressed in Escherichia coli. The secondary structure of RBD was determined using 1H and 13C chemical shift indices and inter- and intrachain nuclear Overhauser enhancements. The secondary structure consists of eight strands in beta-conformation and one alpha-helix, which together comprise 44% of the protein. The beta-strands form three regions of antiparallel beta-sheet. The two lysines previously identified as being critical for receptor binding are located in (Lys1374), and immediately adjacent to (Lys1370) the alpha-helix, which also contains an (Arg1378). Secondary structure predictions of other alpha macroglobulins show the conservation of this alpha-helix and suggest an important role for this helix and for basic residues within it for receptor binding. PMID- 9865956 TI - Self-organizing tree-growing network for the classification of protein sequences. AB - The self-organizing tree algorithm (SOTA) was recently introduced to construct phylogenetic trees from biological sequences, based on the principles of Kohonen's self-organizing maps and on Fritzke's growing cell structures. SOTA is designed in such a way that the generation of new nodes can be stopped when the sequences assigned to a node are already above a certain similarity threshold. In this way a phylogenetic tree resolved at a high taxonomic level can be obtained. This capability is especially useful to classify sets of diversified sequences. SOTA was originally designed to analyze pre-aligned sequences. It is now adapted to be able to analyze patterns associated to the frequency of residues along a sequence, such as protein dipeptide composition and other n-gram compositions. In this work we show that the algorithm applied to these data is able to not only successfully construct phylogenetic trees of protein families, such as cytochrome c, triosephophate isomerase, and hemoglobin alpha chains, but also classify very diversified sequence data sets, such as a mixture of interleukins and their receptors. PMID- 9865957 TI - Probing the ligand binding domain of the GluR2 receptor by proteolysis and deletion mutagenesis defines domain boundaries and yields a crystallizable construct. AB - Ionotropic glutamate receptors constitute an important family of ligand-gated ion channels for which there is little biochemical or structural data. Here we probe the domain structure and boundaries of the ligand binding domain of the AMPA sensitive GluR2 receptor by limited proteolysis and deletion mutagenesis. To identify the proteolytic fragments, Maldi mass spectrometry and N-terminal amino acid sequencing were employed. Trypsin digestion of HS1S2 (Chen GQ, Gouaux E. 1997. Proc Natl Acad Sci USA 94:13431-13436) in the presence and absence of glutamate showed that the ligand stabilized the S1 and S2 fragments against complete digestion. Using limited proteolysis and multiple sequence alignments of glutamate receptors as guides, nine constructs were made, folded, and screened for ligand binding activity. From this screen, the S1S21 construct proved to be trypsin- and chymotrypsin-resistant, stable to storage at 4 degrees C, and amenable to three-dimensional crystal formation. The HS1S21 variant was readily prepared on a large scale, the His tag was easily removed by trypsin, and crystals were produced that diffracted to beyond 1.5 A resolution. These experiments, for the first time, pave the way to economical overproduction of the ligand binding domains of glutamate receptors and more accurately map the boundaries of the ligand binding domain. PMID- 9865959 TI - Effect of denaturant and protein concentrations upon protein refolding and aggregation: a simple lattice model. AB - We present a study of the competition between protein refolding and aggregation for simple lattice model proteins. The effect of solvent conditions (i.e., the denaturant concentration and the protein concentration) on the folding and aggregation behavior of a system of simple, two-dimensional lattice protein molecules has been investigated via (dynamic Monte Carlo simulations. The population profiles and aggregation propensities of the nine most populated intermediate configurations exhibit a complex dependence on the solution conditions that can be understood by considering the competition between intra- and interchain interactions. Some of these configurations are not even seen in isolated chain simulations; they are observed to be highly aggregation prone and are stabilized primarily by the aggregation reaction in multiple-chain systems. Aggregation arises from the association of partially folded intermediates rather than from the association of denatured random-coil states. The aggregation reaction dominates over the folding reaction at high protein concentration and low denaturant concentration, resulting in low refolding yields at those conditions. However, optimum folding conditions exist at which the refolding yield is a maximum, in agreement with some experimental observations. PMID- 9865960 TI - Selective and asymmetric action of trypsin on the dimeric forms of seminal RNase. AB - Dimeric seminal RNase (BS-RNase) is an equilibrium mixture of conformationally different quaternary structures, one characterized by the interchange between subunits of their N-terminal ends (the MXM form); the other with no interchange (the M=M form). Controlled tryptic digestion of each isolated quaternary form generates, as limit digest products, folded and enzymatically active molecules, very resistant to further tryptic degradation. Electrospray mass spectrometric analyses and N-terminal sequence determinations indicate that trypsin can discriminate between the conformationally different quaternary structures of seminal RNase, and exerts a differential and asymmetric action on the two dimeric forms, depending on the original quaternary conformation of each form. The two digestion products from the MXM and the M=M dimeric forms have different structures, which are reminiscent of the original quaternary conformation of the dimers: one with interchange, the other with no interchange, of the N-terminal ends. The surprising resistance of these tryptic products to further tryptic action is explained by the persistence in each digestion product of the original intersubunit interface. PMID- 9865958 TI - Unfolding and refolding of dimeric creatine kinase equilibrium and kinetic studies. AB - Equilibrium and kinetic studies of the guanidine hydrochloride induced unfolding refolding of dimeric cytoplasmic creatine kinase have been monitored by intrinsic fluorescence, far ultraviolet circular dichroism, and 1-anilinonaphthalene-8 sulfonate binding. The GuHCl induced equilibrium-unfolding curve shows two transitions, indicating the presence of at least one stable equilibrium intermediate in GuHCl solutions of moderate concentrations. This intermediate is an inactive monomer with all of the thiol groups exposed. The thermodynamic parameters obtained by analysis using a three-state model indicate that this intermediate is similar in energy to the fully unfolded state. There is a burst phase in the refolding kinetics due to formation of an intermediate within the dead time of mixing (15 ms) in the stopped-flow apparatus. Further refolding to the native state after the burst phase follows biphasic kinetics. The properties of the burst phase and equilibrium intermediates were studied and compared. The results indicate that these intermediates are similar in some respects, but different in others. Both are characterized by pronounced secondary structure, compact globularity, exposed hydrophobic surface area, and the absence of rigid side-chain packing, resembling the "molten globule" state. However, the burst phase intermediate shows more secondary structure, more exposed hydrophobic surface area, and more flexible side-chain packing than the equilibrium intermediate. Following the burst phase, there is a fast phase corresponding to folding of the monomer to a compact conformation. This is followed by rapid assembly to form the dimer. Neither of the equilibrium unfolding transitions are protein concentration dependent. The refolding kinetics are also not concentration dependent. This suggests that association of the subunits is not rate limiting for refolding, and that under equilibrium conditions, dissociation occurs in the region between the two unfolding transitions. Based upon the above results, schemes of unfolding and refolding of creatine kinase are proposed. PMID- 9865961 TI - Quantum chemical calculations of the reorganization energy of blue-copper proteins. AB - The inner-sphere reorganization energy for several copper complexes related to the active site in blue-copper protein has been calculated with the density functional B3LYP method. The best model of the blue-copper proteins, Cu(Im)2(SCH3)(S(CH3)2)(0/+), has a self-exchange inner-sphere reorganization energy of 62 kJ/mol, which is at least 120 kJ/mol lower than for Cu(H2O)4(+/2+). This lowering of the reorganization energy is caused by the soft ligands in the blue-copper site, especially the cysteine thiolate and the methionine thioether groups. Soft ligands both make the potential surfaces of the complexes flatter and give rise to oxidized structures that are quite close to a tetrahedron (rather than tetragonal). Approximately half of the reorganization energy originates from changes in the copper-ligand bond lengths and half of this contribution comes from the Cu-S(Cys) bond. A tetragonal site, which is present in the rhombic type 1 blue-copper proteins, has a slightly higher (16 kJ/mol) inner-sphere reorganization energy than a trigonal site, present in the axial type 1 copper proteins. A site with the methionine ligand replaced by an amide group, as in stellacyanin, has an even higher reorganization energy, about 90 kJ/mol. PMID- 9865962 TI - Solution structure of the His12 --> Cys mutant of the N-terminal zinc binding domain of HIV-1 integrase complexed to cadmium. AB - The solution structure of His12 --> Cys mutant of the N-terminal zinc binding domain (residues 1-55; IN(1-55)) of HIV-1 integrase complexed to cadmium has been solved by multidimensional heteronuclear NMR spectroscopy. The overall structure is very similar to that of the wild-type N-terminal domain complexed to zinc. In contrast to the wild-type domain, however, which exists in two interconverting conformational states arising from different modes of coordination of the two histidine side chains to the metal, the cadmium complex of the His12 --> Cys mutant exists in only a single form at low pH. The conformation of the polypeptide chain encompassing residues 10-18 is intermediate between the two forms of the wild-type complex. PMID- 9865963 TI - Ca2+-dependent conformational changes in bovine GCAP-2. AB - GCAP-2, a mammalian photoreceptor-specific protein, is a Ca2+-dependent regulator of the retinal membrane guanylyl cyclases (Ret-GCs). Sensing the fall in intracellular free Ca2+ after photo-excitation, GCAP-2 stimulates the activity of Ret-GC leading to cGMP production. Like other members of the recoverin superfamily, GCAP-2 is a small N-myristoylated protein containing four EF-hand consensus motifs. In this study, we demonstrate that like recoverin and neurocalcin, GCAP-2 alters its conformation in response to Ca2+-binding as measured by a Ca2+-dependent change in its far UV CD spectrum. Differences in the conformation of the Ca2+-bound and Ca2+-free forms of GCAP-2 were also observed by examining their relative susceptibility to V8 protease. In contrast to recoverin, we do not observe proteolytic cleavage of the myristoylated N-terminus of Ca2+-bound GCAP-2. NMR spectra also show that, in contrast to recoverin, the chemical environment of the N-terminus of GCAP-2 is not dramatically altered by Ca2+ binding. Despite the similarity of GCAP-2 and recoverin, the structural consequences of Ca2+-binding for these two proteins are significantly dissimilar. PMID- 9865964 TI - A solution SAXS study of Borrelia burgdorferi OspA, a protein containing a single layer beta-sheet. AB - The crystal structure of a soluble form of Borrelia burgdorferi outer surface protein A (OspA) complexed with the Fab fragment of a monoclonal antibody has revealed an unusual structure that has a repetitive antiparallel beta topology with a nonglobular, single layer beta-sheet connecting the globular N- and C terminal domains. Earlier NMR studies have shown that the local structure of OspA including the single layer beta-sheet is similar to the crystal structure. Here we report a small angle X-ray scattering (SAXS) study of the global conformation of OspA in solution. The radius of gyration (Rg) and the length distribution function (P(r)) of OspA measured by SAXS in solution are nearly identical to the calculated ones from the crystal structure, respectively. The NMR and SAXS experiments complement each other to show that OspA including the central single layer beta-sheet is a stable structure in solution, and that the OspA crystal structure represents the predominant solution conformation of the protein. PMID- 9865965 TI - Yeast methionine aminopeptidase I can utilize either Zn2+ or Co2+ as a cofactor: a case of mistaken identity? AB - Yeast methionine aminopeptidase I (MetAP I) is one of two enzymes in Saccharomyces cerevisiae that is responsible for cotranslational cleavage of initiator methionines. It has previously been classified as a Co2+ metalloprotease in all prokaryotic and eukaryotic forms studied. However, treatment of recombinant apo-MetAP I with 12.5 microM Zn2+ produces an enzyme that is as active as that reconstituted with 200 microM Co2+. In the presence of physiological concentrations of reduced glutathione (GSH), Co-MetAP I is inactive, while the activity of Zn-MetAP I is increased more than 1.7-fold over Zn-MetAP I assayed in the absence of GSH. Given that the in vivo concentration of Zn2+ is at least 1,000-fold higher than that of Co2+, and that Co2+ is insoluble in physiological concentrations of GSH, it is probable that yeast MetAP I is actually a Zn2+ metalloprotease. Furthermore, unless there are extraordinary conditions that insulate or sequester them from this reducing milieu, that have yet to be identified, there are not likely to be any cytoplasmic enzymes that use free Co2+. PMID- 9865966 TI - The trail to superoxide dismutase. PMID- 9865967 TI - Hormone-inducible expression of secreted factors in zebrafish embryos. AB - The study of gene function at later stages of embryonic development by overexpression experiments is often complicated by genes exerting different functions at multiple stages of development, which renders analysis of stage specific effects difficult. To address this problem an inducible expression system that supports timed expression of essentially any protein, including secreted proteins was designed. The system is based on a two step mechanism. A glucocorticoid inducible, Gal4-site binding chimeric transcription factor is expressed ubiquitously, whereas a gene of interest is placed under the control of a Gal4-site driven promoter. Treatment of zebrafish embryos injected with such constructs with the synthetic glucocorticoid dexamethasone results in readily detectable reporter activity within 3 h. The system was tested with induced expression of Xactivin(beta)B and X(wnt), which both were shown to induce morphological abnormalities, as well as alterations in the expression patterns of goosecoid and otx2, respectively. Coinjection of an inducible lacZ reporter vector served as an indicator for expressing cells in embryos. The present results demonstrate that this is a versatile inducible expression system for use in vertebrate embryos, that also supports expression of secreted proteins. PMID- 9865968 TI - Histone deacetylase mRNA temporally and spatially regulated in its expression in sea urchin embryos. AB - SpHDAC1, a cDNA homolog of the yeast Rpd3 and higher eukaryotic histone deacetylases (HDAC), was cloned from the sea urchin Strongylocentrotus purpuratus. Its predicted polypeptide and the Rpd3 homologs were highly identical in two-thirds of their lengths, but diverged in their carboxyl-terminal regions in both length and sequence. SpHDAC1 transcripts, which reached maximal concentration at the blastula stages, and diminished thereafter, were neither ubiquitously expressed nor restricted to particular cell lineages, but appeared successively in distinct embryonic regions. In the blastula, transcripts were concentrated in a ring within the vegetal plate, comprising primordial endoderm, and, at the outset of gastrulation, in primordial hindgut endoderm. However, in early to mid-gastrula transcripts, they also appeared in oral ectoderm. In the late-stage gastrula, expression developed in the foregut. These shifts in spatial expression, together with an observed developmental blockage prior to sea urchin gastrulation by the histone deacetylase inhibitor trichostatin A, suggest a stepwise involvement of SpHDAC1 gene expression or SpHDAC1 functionality in the events of normal gastrulation. PMID- 9865969 TI - Expression pattern analysis of SGF-3/POU-M1 in relation to sericin-1 gene expression in the silk gland. AB - Embryonic and larval expression patterns of the sericin-1 gene and its presumed transcription factor, SGF-3/POU-M1, in the silk gland were analyzed by in situ hybridization and immunohistochemistry. The sericin-1 transcripts were first detected at embryonic stage 26 in an increasing gradient pattern in the middle and posterior part of the middle silk gland (MSG), while at the same stage the SGF-3/POU-M1 was already present in the entire anterior silk gland (ASG) and in the MSG but with a decreasing gradient pattern. The latter expression pattern was consistently maintained through all larval stages, while the sericin-1 expression was detected during the feeding stages but disappeared at the molting stages. These observations suggest that, although the SGF-3/POU-M1 was proposed to be a positive transcription factor for the sericin-1 gene, the protein might function in a negative manner on sericin-1 gene transcription. Alternatively, it is also possible that the sericin-1 gene might require another unidentified factor or mediator for in vivo transcription. PMID- 9865970 TI - Cloning of cDNA for newt WT1 and the differential expression during spermatogenesis of the Japanese newt, Cynops pyrrhogaster. AB - To investigate the function of Wilms' tumor 1 (WT1) during spermatogenesis, cDNA for newt WT1 homolog was cloned and the expression of WT1 in newt testes was examined. The cDNA is 2089 bp in length and encodes 426 amino acid (aa) residues. The deduced aa sequence shares 76 and 79% homology with human and Xenopus WT1, respectively. Northern blot analysis shows that WT1 mRNA, 3.2 and 4.5 kb in length, are expressed in the testis and kidney. Both WT1 mRNA species are detected in various stages of spermatogenesis, but the 3.2 kb mRNA is highly expressed in spermatogonia and mature sperm stages, while the amount of 4.5 kb mRNA is almost constant throughout spermatogenesis. In situ hybridization reveals that WT1 mRNA is localized in Sertoli cells. Moreover, immunohistochemical analysis shows that WT1 protein is highly expressed in the nuclei of Sertoli cells in early spermatogonia and mature sperm stages, but not in pericystic cells or germ cells. These results suggest that WT1 is involved in the regulation of gene expression in Sertoli cells, depending on the spermatogenic stage. PMID- 9865972 TI - Behavior of centrioles during meiosis in the male silkworm, Bombyx mori (Lepidoptera). AB - The behavior of centrioles during eupyrene and apyrene meiosis was examined in the silkworm, Bombyx mori, by transmission electron microscopy and indirect immunofluorescence for tubulin. In eupyrene spermatocytes the centrioles, accompanied by axonemes, attached temporarily to the nucleus at diplotene, then detached from the nucleus in diakinesis. After the separation, a beret-shaped structure consisting of a double membrane covered the proximal region of the pair of centrioles. The structure disappeared after breakdown of the nuclear membrane. The centriole, with the axoneme, reattached to the nucleus at telophase I. The process was repeated during meiosis II until the centrioles maintained their nuclear attachment in newly developed spermatids. In stark contrast to their eupyrene counterparts, apyrene spermatocytes were conspicuously devoid of any attachment of the centrioles to the nucleus. These eupyrene-specific and apyrene specific relationships were consistently and repeatedly found between the nuclear membrane and centrioles, giving rise to suspicion that the behavioral phenomena may be related to differentiation of the dimorphic sperm types. PMID- 9865971 TI - Role of cell adhesion in the specification of pigment cell lineage in embryos of the sea urchin, Hemicentrotus pulcherrimus. AB - To clarify the role of cell adhesion in the specification of pigment cell lineage in sea urchin embryos, cell contacts were inhibited by Ca2+-free artificial seawater (ASW) treatment, and the number of differentiated pigment cells was examined by the method devised for the present study. Obtained results showed that inhibition of cell contacts during mid-to-late blastula stage greatly affects the number of pigment cells. Treatment with Ca2+-free ASW during 7.5-10.5 h of development drastically decreased the number of pigment cells, indicating that cell adhesion during this period is indispensable for the specification of pigment cell lineage. On the other hand, the number of pigment cells were increased by the treatment during 9.5 12.5 h of development. It was suggested that this increase was caused by excess divisions of the precursor cells, that is, the division schedule of the precursor cells was altered by inhibition of cell contacts at this period. Interestingly, the number of pigment cells was a multiple of four in a majority of embryos in which pigment cells were drastically decreased in number. These findings suggest that the founder blastomeres of the pigment cell lineage are specified during 7-10 h of development, and that these blastomeres divide twice before they differentiate into pigment cells. PMID- 9865973 TI - An alternatively spliced gene encoding a Y-box protein showing maternal expression and tissue-specific zygotic expression in the ascidian embryo. AB - An ascidian Y-box protein gene was cloned, designated as CiYB, which consists of a highly conserved cold shock domain and an auxiliary tail domain with alternating modules of acidic and basic amino acids. CiYB is a single copy gene in the ascidian genome. During oogenesis and early development, CiYB produces three different transcripts (CiYB1, CiYB2 and CiYB3) by alternate splicing. CiYB1 and CiYB2 were expressed during oogenesis, suggesting that they are recruited into maternal ribonucleoprotein particles. According to gel mobility shift assay, the CiYB1 protein has the ability to bind RNA. The sequence preference of RNA binding is similar to that of the Xenopus Y-box protein (FRGY2), which is a major component of the maternal messenger ribonucleoprotein particles (mRNP) in the oocyte. These results suggest that the ascidian Y-box protein may have an important role for masking and translational regulation of maternal mRNA. Furthermore, CiYB1, CiYB2 and CiYB3 were expressed zygotically in a tissue restricted manner. CiYB1 was expressed specifically in muscle precursor blastomeres and tail muscle cells suggesting its important role in muscle differentiation. PMID- 9865974 TI - Primary structure of a 120 kDa protein associated with the fucose sulfate glycoconjugate constituting the acrosome reaction-inducing substance of the sea urchin, Hemicentrotus pulcherrimus. AB - A fucose sulfate glycoconjugate (FSG), a natural acrosome reaction-inducer, was purified from the egg jelly of the sea urchin Hemicentrotus pulcherrimus. The FSG is composed primarily of four constituents: a 120 kDa protein, a 237 kDa protein, a 258 kDa protein, and a polysaccharide-containing protein. Among them, the 120 kDa protein was thought to play a critical role in the association of other FSG constituent proteins, and therefore was characterized from a structural point of view. The protein was isolated from the carboxymethylated FSG by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, and then digested with trypsin to obtain information regarding the primary structure. Based on the partial amino acid sequences of three internal peptides (FSG120KA: LHNNEYGYGDTAAGEPELAQEEID, FSG 120KG: AIDIPAETGHYGR, and FSG120KC: RPTFDLADAVDT) and the N-terminal peptide (LHNNEYGYGDTAAGEPELAQQEID) of the 120 kDa protein obtained from intact FSG, degenerate oligonucleotide primers were synthesized and used to amplify a 297 bp cDNA fragment. This fragment enabled us to obtain the full-length cDNA (3176 bp) by 5'- and 3'-rapid amplification of cDNA ends. The deduced amino acid sequence revealed that the 120 kDa protein is composed of 663 amino acid residues including 72 cysteine residues, and hence, about 40% is presumed to be carbohydrate by weight. The 120 kDa protein plays an important role in the association of FSG constituent proteins (258 and 237 kDa) through disulfide bonds. PMID- 9865975 TI - Developmental studies for identification of the inhibitory center of melanotropes in the toad, Bufo japonicus. AB - Two series of experiments were performed to identify the inhibitory center of the melanotropes in the intermediate lobe of hypophysis of the toad, Bufo japonicus. First, developmental changes in the distribution of dopaminergic neurons were examined from hatching stage to postmetamorphosis using an antiserum against dopamine synthase (tyrosine hydroxylase, TH). In the postmetamorphic toads, TH positive cell bodies were localized in three clusters. One was the preoptic recess organ (PRO) in the prechiasmatic area, the other two were the paraventricular organ (PVO) and infundibular nucleus (IN) in the postchiasmatic area. Each of them exhibited different ontogenetic changes. During larval development, TH-positive cell bodies were first detected in the PVO and IN at a premetamorphic stage. The number of immunoreactive cells increased rapidly in both loci as metamorphosis proceeded, although the two nuclei showed different growth profiles. By contrast, in the PRO, a very small number of immunoreactive cells were observed before the onset of the prometamorphic period. Although the number of immunoreactive neurons increased as metamorphosis progressed, early neurons were confined to the caudal area of the PRO (cPRO), the rostral area of the PRO (rPRO) being devoid of TH-positive cells. Immunoreactive TH neurons appeared in the rPRO for the first time at the end of metamorphic climax. This timing coincided well with the development of TH-positive nerve endings in the pars intermedia (PI) and median eminence. In the second series of experiments, the embryonic primordium of the PRO was surgically extirpated from open neurulae to examine the effects of PRO-ectomy. In 75% of the operated animals, background adaptation was not observed, their dermal melanophores remained permanently dispersed even on the white background. Dopaminergic neurons in the rPRO and the immunoreactive nerve endings in the PI and median eminence were scarcely observed in these animals. It was concluded that the present data strongly support the hypothesis that rPRO is the center of white-background adaptation. PMID- 9865976 TI - Cysteine proteinase plays a key role for the initiation of yolk digestion during development of Xenopus laevis. AB - In electrophoretic analyses, extracts of Xenopus laevis neurulae exhibited activities digesting yolk proteins maximally at pH 4.8. These activities were completely inhibited by a mixture of pepstatin A and Z-Phe-Phe-CHN2, thus being identifiable as cathepsin D and cysteine proteinase. The electrophoretic profiles of yolk proteins cleaved by embryonic extracts changed at gastrula stages; the profile before stage 13 was the same as that given by cathepsin D treatment and the profile at stage 13 was a combination of the profile given by cathepsin D treatment and that given by cysteine proteinase treatment. Quantitative measurement of enzyme activities showed that the cathepsin D activity that was preserved from the beginning of development increased from stages 13 to 25 and decreased thereafter, whereas the cysteine proteinase activity appeared at stage 13, gradually increased until stage 35 and strongly increased thereafter. Immunoblot analyses showed that the 43 kDa form of cathepsin D was processed to its 36 kDa form, presumably by cysteine proteinase. This change can explain the increase of cathepsin D activity at stage 13 and thereafter. Immunofluorescent staining with the antibody against cysteine proteinase occurred in mesodermal and ectodermal cells other than neural ones at stages 13-24, and in the endodermal cells at stages 24-36. Faint staining in the neural ectoderm persisted from stages 18 to 36. Immunoelectron microscope observation showed that what stained was the superficial layer of yolk platelets. All these results indicate that cysteine proteinase plays a key role in the initiation of yolk digestion during embryonic development. PMID- 9865977 TI - Detection of phospholipase Cgamma in sea urchin eggs. AB - Phosphorylation on tyrosine and turnover of polyphosphoinositide metabolism are rapidly stimulated after fertilization. However, the interconnection between these pathways remains to be determined. In the present paper it is demonstrated that eggs of two different sea urchin species contain tyrosine phosphorylated proteins with calcium-sensitive phospholipase C activity. We have investigated whether phospholipase Cgamma (PLCgamma), characteristic of tyrosine kinase receptors, could be responsible for this activity. Western blot and immunocytochemistry performed with antibodies directed against PLCgamma revealed the presence of this protein in cortical regions. It was also observed that PLCgamma displayed calcium-sensitive activity. The present results suggest that PLCgamma may be part of the cascade of events leading to the calcium signal responsible for egg activation at fertilization. PMID- 9865978 TI - Overexpression of CAF1 encoding a novel Ca2+-binding protein stimulates the transition of Dictyostelium cells from growth to differentiation. AB - Among the expressed genes associated with the switch-over of Dictyostelium cells from cell proliferation to differentiation, the Calfumirin-1 (CAF1) gene has been shown to be preferentially expressed at the initial step of differentiation, encoding a novel Ca2+-binding protein (Abe & Maeda 1995). To analyze precisely the function of CAF1, transformants overexpressing the CAF1 mRNA at the vegetative growth phase and also CAF1-null mutants were prepared, and their developmental features were compared with those of parental wild-type cells. As a result, the CAF1-overexpression was found to promote cell differentiation, possibly through prompt induction of the cAMP receptor 1 (CAR1) gene expression. In addition, the CAF1-overexpressing cells were able to differentiate even under low external Ca2+ ([Ca2+]e) conditions around 10(-6) mol/L at which non transformed wild-type cells never differentiated. Unexpectedly, however, the CAF1 null mutant produced by homologous recombination exhibited apparently normal development to form fruiting bodies on non-nutrient agar. These results seem to indicate that CAF1-overexpression has a stimulatory effect on differentiation, but that the CAF1 protein is not necessarily required for the phase-shift of cells from growth to differentiation. PMID- 9865979 TI - Delay in administering the first dose of antibiotics in patients admitted to hospital with serious infections. AB - The interval from the time of admission to the emergency room until the administration of antibiotics in patients presenting with a serious infectious disease was analysed. Fifty patients presumptively diagnosed in the emergency room as having a serious infection (respiratory tract, urinary tract, erysipelas, fever with neutropenia or bacteremia) needing immediate empirical antibiotic treatment were enrolled in the study. A median interval from time of admission to administration of antibiotics of 5 hours was determined (range 0.6-13.3 h). The interval was significantly shorter in patients admitted at night than in patients admitted during office hours (3.7 vs. 6.0 h, P< 0.05). There was no difference with respect to the presenting features, body temperature, laboratory values at presentation or number of cultures performed. In 41 of the 50 patients blood samples were taken for culture. More than 80% of the patients received an antibiotic chosen in accordance with hospital guidelines. The analysis revealed that the median delay of 5 hours before patients received their initial dose of antibiotic depended on several factors. Attempts to provide optimal antimicrobial therapy should thus concentrate not only on the correct choice and dosage of a drug but also on prompt institution of therapy. PMID- 9865980 TI - Etiology of respiratory tract infection in adults in a general practice setting. AB - A prospective study was conducted over a 3-month winter period in three general practice clinics in an urban population in southern Israel to identify the etiological agents of respiratory tract infections (RTI) in adults. RTI was defined as an acute febrile illness with cough, coryza, sore throat or hoarseness. Serum samples were taken from all patients in both the acute and convalescent phases of their illness. Tests were conducted for detection of 17 microorganisms known to cause RTI, including serological tests for 16 known pathogens. An etiological diagnosis was established in 80 (66%) of the 122 patients who participated in the study. The distribution of the etiological agents was as follows: influenza B virus in 27 (22%) patients. Chlamydia pneumoniae in 22 (18%), Legionella spp. in 15 (12%), Mycoplasma pneumoniae in 13 (11%), influenza A virus in 11 (9%), Bordetella pertussis in 9 (7%), adenovirus in 4, Epstein Barr virus in 4, Haemophilus influenzae in 3, beta-hemolytic streptococci in 3, Streptococcus pneumoniae in 2, respiratory syncytial virus in 2, parainfluenza 1 virus in 2 and parainfluenza 2 virus in 1. No patients were found to be infected with Coxiella burnetii, Moraxella catarrhalis or parainfluenza 3 virus. More than one pathogen was identified in 27 (34%) patients in whom an etiological diagnosis was established. It is concluded that RTI is caused by a broad spectrum of etiological agents, a considerable number of patients having evidence of infection with more than one pathogen. The therapeutic significance of these findings should be elucidated in further studies. PMID- 9865981 TI - Trend in incidence and case fatality of meningococcal disease over 16 years in Northern Denmark. AB - The incidence and case fatality rates of meningococcal disease were assessed in the county of Northern Jutland, Denmark, during the 16-year period from 1980 to 1995. A total of 320 patients were identified from the Meningococcal Research Database, which comprises information from the following sources: (i) the Department of Public Health, to whom notification of meningococcal disease is obligatory; (ii) the Regional Hospital Discharge Registry; and (iii) the register of the regional department of clinical microbiology. In order to assess prognostic indicators assessable at admission, information was collected for each patient from hospital records regarding contacts, symptoms and signs on arrival, laboratory data, and course of disease. The mean incidence was 4.3 cases per 100000 persons per year (range, 2.7-7.7). The incidence increased slightly during the period studied. Overall, the case fatality rate was 9.7%, with a significant rise occurring during the period (P=0.016) and a peak occurring in 1992. Advanced age (> or = 50 years), seizures, impaired consciousness, and skin bleeding on arrival at hospital were predictors of death. PMID- 9865982 TI - Epidemiology of nosocomial bloodstream infections in Belgium, 1992-1996. AB - The main results of the bloodstream infection (BSI) component of the Belgian National Programme for the Surveillance of Hospital Infections (NSIH project) are reported. From October 1992 to September 1996, 117 hospitals (59.1% of Belgian acute-care institutions) reported 13678 nosocomial BSIs. The incidence was 7.05 BSI episodes per 10000 patient-days. The incidence of BSI increased with hospital size and over time. Bloodstream infections were secondary to an infectious body site in 40.3% of the episodes, catheter-related in 23.5%, and of unknown origin in 36.2%. The associated in-hospital mortality was 31.4% and was highest in BSIs secondary to a respiratory tract infection (49.3%). In intensive care units, the incidence of BSI was 38.5 per 10000 patient-days. Coagulase-negative staphylococci were the most prevalent microorganisms (22%), followed by Staphylococcus aureus (14.1%) and Escherichia coli (13.5%). In catheter-related BSIs, these proportions were 41.9%, 18.8%, and 2.3%, respectively. The proportion of polymicrobial episodes was 9.9%. Methicillin resistance in Staphylococcus aureus was 22.3%. With its high participation rate, the NSIH project has characterized the epidemiology of nosocomial BSIs in Belgium during the period studied. PMID- 9865983 TI - Factors associated with improved outcome of Pseudomonas aeruginosa bacteremia in a Finnish university hospital. AB - All 134 episodes of bacteremia caused solely by Pseudomonas aeruginosa in a university hospital in the periods 1976-1982 and 1992-1996 were reviewed retrospectively to determine the clinical manifestations, outcome and prognostic factors. The mortality for the 30-day interval after drawing the first positive blood culture was 41%, but dropped from 53% in the first period to 29% in the second period (P=0.006). Mortality was highest in patients treated with an aminoglycoside only, as against those treated with other appropriate antibiotics (55% versus 25%, P=0.001). Over the two decades studied, use of an aminoglycoside only decreased, use of paracetamol (=acetaminophen) increased, and removal of both urinary and blood vessel catheters became more common. The mortality was 18% in patients with catheter removal (46% in the other patients, P=0.017) and 27% in patients who received paracetamol around the time of drawing the first positive blood culture (50% for the other patients, P=0.010). Logistic regression analysis showed that shock, central nervous system involvement, preceding thromboembolism and rapidly fatal underlying disease were associated with a fatal outcome, whereas catheter removal, appropriate antibiotic therapy and paracetamol therapy were associated with survival. The improved prognosis of Pseudomonas aeruginosa bacteremia over the two decades is thus due mainly to three changes in management of the infection: the more frequent use of new anti-pseudomonal beta-lactams and ciprofloxacin instead of aminoglycosides as monotherapy; the more frequent practice of removing catheters; and the increased use of paracetamol around the time of drawing the first positive blood sample. PMID- 9865984 TI - Influence of hepatitis G virus coinfection on the clinical course of chronic hepatitis C. AB - Hepatitis G virus (HGV) is a parenterally transmitted virus, frequently associated with hepatitis C virus infection. Hepatitis G virus RNA was detected by reverse transcription-polymerase chain reaction in the serum of 40 patients with chronic hepatitis C. Nine (22.5%) patients had evidence of hepatitis G virus viraemia. No significant epidemiological or virological differences could be demonstrated between subjects infected with both hepatitis G virus and hepatitis C virus and subjects infected with hepatitis C virus alone. Aminotransferase values were comparable between the two groups, whereas higher levels of cholestatic enzymes (P< 0.001) were reported in the hepatitis G virus/hepatitis C virus-positive patients. A liver biopsy was performed on all 40 patients no later than 6 months before recruitment. The mean histological activity index did not differ between hepatitis G virus-positive and hepatitis G virus-negative patients, whereas specific histological features such as macrovesicular steatosis, portal granulomas, and bile duct damage were more commonly observed among the coinfected patients. The results indicate that coinfection with hepatitis G virus probably does not have a significant effect on hepatitis C virus-induced hepatic damage. PMID- 9865985 TI - Comparison of oral cefixime and intravenous ceftriaxone followed by oral amoxicillin in disseminated Lyme borreliosis. AB - Two treatment regimens for disseminated Lyme borreliosis (mainly neurologic and musculoskeletal manifestations) were compared in a randomized trial. A group of 30 patients received oral cefixime 200 mg combined with probenecid 500 mg three times daily for 100 days. Another group of 30 patients received intravenous ceftriaxone 2 g daily for 14 days followed by oral amoxicillin 500 mg combined with probenecid 500 mg three times daily for 100 days. There was no statistically significant difference in the outcome of infection between the two groups. However, the total number of patients with relapses or no response at all and the number of positive polymerase chain reaction findings after therapy were greater in the cefixime group. The general outcomes of infection in patients with disseminated Lyme borreliosis after 3-4 months of therapy indicate that prolonged courses of antibiotics may be beneficial in this setting, since 90% of the patients showed excellent or good treatment responses. PMID- 9865986 TI - Incidence of Chlamydia pneumoniae infection in vertically HIV-1 infected children. AB - The rate of seroconversion for antibody to Chlamydia pneumoniae was analysed in blood samples of 26 vertically HIV-1 infected children and 14 seroreverter children (HIV-negative children born to HIV-positive mothers) during a 3-year study period. Seroconversion for Chlamydia pneumoniae was found in 13 of 26 HIV-1 infected children and in 1 of 14 in the seroreverter group (P=0.013). A lower mean CD4+ cell count and p24 antigen positivity at enrolment were significantly associated with seroconversion for Chlamydia pneumoniae. Signs and symptoms of acute respiratory infection were recorded in the 30 to 40 days preceding collection of the blood samples showing seroconversion for Chlamydia pneumoniae in 8 of 13 HIV-1 infected children and in the single seroreverter. This study confirms the potential role of Chlamydia pneumoniae in the pathogenesis of respiratory tract infections in HIV-1 infected subjects. PMID- 9865987 TI - Investigation of an outbreak of wound infections due to Alcaligenes xylosoxidans transmitted by chlorhexidine in a burns unit. AB - Alcaligenes xylosoxidans, an environmental gram-negative bacillus, was isolated within a 1-month period from six patients in a pediatric burns unit. Twelve isolates were studied, one from each of the six patients (five from wound cultures and one from a blood culture) and one from each of six contaminated atomizers containing chlorhexidine diluted to 600 mg/l. The biochemical and susceptibility patterns of all the isolates were similar, and their DNA enzyme restriction patterns were identical. The epidemic strain of Alcaligenes xylosoxidans was probably introduced into the atomizers during handling of the diluted solution, which failed to eliminate it. PMID- 9865989 TI - Effect of the addition of vancomycin on the performance of an automated nonradioactive system for detection of mycobacteria. AB - A recently developed automated, nonradioactive system for the detection of mycobacteria (MB/BacT; Organon Teknika, Belgium) has provided good results, but the contamination rate was found to be higher than that obtained with the radiometric Bactec 460 system (Becton Dickinson, USA). In the present study, the effects of adding vancomycin (1 microg/ml) to the antibiotic mixture of the nonradioactive system were evaluated, and the performance of the system with versus without vancomycin was compared. Three hundred sputum samples were tested, using the radiometric system as the reference method. Mycobacteria were isolated from 47 (15.7%) samples. The nonradioactive system with and without vancomycin detected 42 and 43 strains, respectively; the time to detection was 1 day shorter with the medium without vancomycin (15.7 days vs. 14.3 days). The radiometric system detected 42 strains of mycobacteria in a mean detection time of 13.6 days. Contamination rates with the nonradioactive system were 6.7% in the medium without vancomycin and 2.7% in the medium with vancomycin. The latter figure was approximately the same as the contamination rate found with the radiometric system (2.3%). Our data suggest that the addition of vancomycin considerably reduces the number of contaminants in the MB/BacT medium without affecting the performance of the system. PMID- 9865988 TI - Utility of serial rectal swab cultures for detection of ceftazidime- and imipenem resistant gram-negative bacilli from patients in the intensive care unit. AB - Forty-four patients receiving intensive care were studied prospectively to assess the utility of serial rectal swab cultures and clinical correlates of resistance for Pseudomonas aeruginosa, Enterobacter spp., Citrobacter spp., Morganella morganii, and Serratia marcescens strains resistant to ceftazidime or imipenem. Strains of Pseudomonas aeruginosa, Enterobacter spp., Citrobacter spp., or Morganella morganii were found in 26 of 44 (59%) patients: 17 (65%) in clinical sites (11 with concomitant rectal isolates) and nine (35%) in a rectal site only. Of 49 total isolates, 13 (26.5%) were resistant: 10 (20.4%) to ceftazidime and three (6.1%) to imipenem. Surveillance rectal swabs from 27 patients without a clinical isolate identified two patients with resistant organisms (15% of all resistant isolates). The majority of resistance to ceftazidime or imipenem among Pseudomonas or Enterobacter can be detected by the use of clinical specimens alone. PMID- 9865990 TI - Successful treatment with ampicillin and fluoroquinolones of human endocarditis due to high-level gentamicin-resistant enterococci. AB - Two cases of endocarditis, one caused by high-level gentamicin-resistant Enterococcus durans and the other by high-level gentamicin- and glycopeptide resistant Enterococcus faecalis. successfully treated with a combination of ampicillin and a fluoroquinolone are reported. Both strains were susceptible to ampicillin. Enterococcus faecalis was susceptible to ciprofloxacin and to ofloxacin, but Enterococcus durans was moderately resistant to these agents. Microbiological and clinical cure was obtained with a 6-week course of ampicillin plus ciprofloxacin in one case and with ofloxacin in the second case due to intolerance to ciprofloxacin. The efficacy of the treatment was predicted in vitro by time-kill studies and by adequate serum bactericidal titres. PMID- 9865991 TI - Rhodococcus equi and Nocardia brasiliensis infection of the brain and liver in a patient with acute nonlymphoblastic leukemia. AB - Resolution of neutropenia is usually followed by resolution of fever in patients with febrile neutropenia. However, in some cases fever continues even when the patient is no longer neutropenic. Described here is a case of acute myeloblastic leukemia complicated by brain abscess, pulmonary nodules, and hepatic lesions. The patient's fever had continued after the neutropenia resolved; brain and hepatic cultures grew Rhodococcus equi and Nocardia brasiliensis. Although Rhodococcus infections occur frequently in patients with HIV infection, they are uncommon in patients with acute leukemia. PMID- 9865992 TI - Two fatal cases of nosocomial Legionella pneumophila pneumonia associated with a contaminated cold water supply. PMID- 9865993 TI - Absence of Pneumocystis carinii carriers among patients with cystic fibrosis. PMID- 9865994 TI - Urethritis due to Streptococcus pyogenes. PMID- 9865995 TI - Microsporidiosis due to Enterocytozoon bieneusi infection as a possible cause of traveller's diarrhea. PMID- 9865996 TI - Disseminated Mycobacterium avium infection after liver transplantation. PMID- 9865997 TI - Isolation of Haemophilus influenzae in Central Asia. PMID- 9865998 TI - Final report of the Technology Transfer Workshop on Breast Cancer Detection, Diagnosis, and Treatment. Washington, D.C., USA. May 1-2, 1997. PMID- 9865999 TI - Twelve-month comparative multicenter study of the TCu 380A and ML 250 intrauterine devices in Bangkok, Thailand. AB - The long-term effectiveness of the copper-bearing intrauterine device (IUD) has been documented. This paper reports 12-month results from a multiyear comparative study of the Copper T (TCu) 380A and Multiload (ML) 250 IUD in Bangkok, Thailand, among 1396 women. Continuation of the assigned IUD was relatively high after 12 months of use, with continuation rates of 90.17 and 87.54 per 100 women, respectively. Whereas the accidental pregnancy rate was higher for the ML 250 IUD than for the TCu 380A IUD (1.0 and 0.2 per 100 women, respectively), this difference was not considered statistically significant (p < 0.69). The rate of IUD expulsion was significantly higher among women using the ML 250 IUD than for women using the TCu 380A IUD (4.61 and 2.40 per 100 women, respectively, p = 0.05). PMID- 9866000 TI - Vaginal misoprostol compared with vaginal gemeprost in termination of second trimester pregnancy. A randomized trial. AB - A prospective randomized trial was conducted in 140 women to compare the efficacy of vaginal gemeprost with vaginal misoprostol for termination of second trimester pregnancy. Women requesting termination of second trimester pregnancy were randomized into two groups. Group A women were given 1 mg vaginal gemeprost every 3 h for a maximum of five doses in the first 24 h, whereas group B women were given 400 micrograms vaginal misoprostol every 3 h for a maximum of five doses in 24 h. The median induction-abortion interval in the vaginal misoprostol group (14.1 h) was significantly shorter than that in the gemeprost group (19.5 h). The percentage of women who achieved successful abortion within 24 h in the misoprostol group (80.0%) was significantly higher than that in the gemeprost group (58.6%). There was no significant difference in the incidence of side effects between the two groups except for diarrhea, which was more common in the gemeprost group. The incidence of fever was more common in the misoprostol group. It is concluded that vaginal misoprostol is more effective than gemeprost in termination of second trimester pregnancy. PMID- 9866001 TI - Sex hormone binding globulin and free levonorgestrel index in the first week after insertion of Norplant implants. AB - Levonorgestrel has an inhibitory effect on sex hormone binding globulin (SHBG). This decrease in SHBG leads to an increase in the free levonorgestrel index (FLI), which has a stronger biological effect. The interaction between serum levels of levonorgestrel and SHBG in long-term users of Norplant implants has been described. This study was designed to understand the same interaction immediately after the insertion of the implants, in a group of 16 women, sampled at 0 and 6 h and at 1, 3, and 7 days after Norplant implant insertion. Peak serum levonorgestrel levels were achieved at 24 h after insertion, remaining stable on day 3 and decreasing by > 10% by day 7. SHBG did not change during the first 24 h, but decreased by 19% and 60% on days 3 and 7, respectively. FLI more than doubled from day 1 to day 7 after insertion. The large decrease in SHBG and doubling of FLI is not followed by a similar reduction in levonorgestrel, which is hard to explain without an increase in the release rate of the steroid from the capsule. PMID- 9866002 TI - Twenty-four month comparison of apolipoproteins A-1, A-II and B in contraceptive implant users (Norplant and Implanon) in Birmingham, United Kingdom. AB - A 2-year prospective, randomized study of 60 implant users was performed to evaluate fasting serial serum apolipoprotein A-I, A-II, and B levels before insertion and every 3 months in users of two contraceptive implants releasing levonorgestrel (Norplant) and 3-ketodesogestrel/etonogestrel (Implanon) in Birmingham, United Kingdom. All changes in serum apolipoprotein concentrations, irrespective of an increase or decrease, were smaller than one standard deviation of the mean concentration at baseline and did not exceed the normal range for the laboratory, thus implying no evidence of an increased risk of coronary heart disease. PMID- 9866004 TI - Reversibility with sodium bicarbonate of styrene maleic anhydride, an intravasal injectable contraceptive, in male rats. AB - A copolymer of styrene and maleic anhydride (SMA) was dissolved in dimethyl sulphoxide (DMSO) and injected into the lumen of the vas deferens of rats. The polymer was retained in the lumen for a period of more than 90 days. Sodium bicarbonate (10%), pH 8.9, was used to flush the polymeric material from the vas deferens lumen. The reversibility and patency of the vas deferens was thus obtained. Sodium bicarbonate proved to be an effective alternative to dimethyl sulphoxide for reversal. PMID- 9866003 TI - Experience of Thai women in Bangkok with Norplant-2 implants. AB - A prospective study of the Norplant-2 contraceptive subdermal implant system was conducted in Bangkok, Thailand. The objective of the study was to evaluate the efficacy, adverse effects, and overall acceptability of Norplant-2 implants. A total of 140 women were enrolled in a 3-year clinical trial. The mean age was 29 years. Of all the acceptors, 70% had completed primary school. The continuation rates at years 1, 2, and 3 were, respectively, 94%, 89%, and 83%. No accidental pregnancies occurred throughout the 3 years of use in this study. Personal reasons were the leading cause for termination of Norplant-2 implant use. The 3 year cumulative termination rate for personal reasons was 7.2%. These personal reasons were divorce, husband having vasectomy, and moving away from the study area. The other leading cause for termination was medical reasons; acne, headache, and pain at the implant site were the complaints. The termination rate for medical reasons in year 3 of the study was 4.6%. Prolonged menstrual flow was the other main reason for termination. The 3-year cumulative termination rate for menstrual irregularities was 3.8%. In this study, the cumulative termination for planned pregnancy at the end of the year 3 was only 1.6%. The incidence of difficult removals was 8%. Breakage of the rods on removal was encountered in the majority of these cases. The study findings presented suggest that the Norplant-2 implants are highly effective with high continuation rates. The Norplant-2 system could become another choice of long acting reversible contraception for Thai women. PMID- 9866005 TI - Women's preferences for vaginal antimicrobial contraceptives. I. Methodology. AB - Sexually transmitted diseases, including AIDS, and unplanned pregnancies continue to be a serious worldwide problem. A number of organizations are developing woman controlled vaginal formulations to prevent these problems. However, little information is available regarding the types of products women prefer even though such knowledge is essential to obtain widespread use. This is the first of several articles that describe the results of a consumer preference study for such vaginal formulations performed in Campinas, Sao Paulo State, Brazil. Because no published methodology was available, the instruments and interview techniques were developed first and procedures established for the identification and participation of research subjects. After preparation of a questionnaire, a pilot study was performed to evaluate it, to establish the interview technique, and to determine the optimal method for subject recruitment. Based on the results, the approach was selected and applied to 635 subjects from different age and socioeconomic groups. The developed methodology and questionnaire, the advantages and the problems encountered, are presented. PMID- 9866006 TI - Women's preferences for vaginal antimicrobial contraceptives. II. Preferred characteristics according to women's age and socioeconomic status. AB - A study was carried out to identify characteristics that women would want for an idealized vaginal contraceptive, and the possible association of these characteristics with age and socioeconomic status. The study was done in Campinas, Sao Paulo State, Brazil. A total of 635 women were selected by age and socioeconomic status, using the "social network" technique. Almost half were adolescents (15-19 years old) and the rest were adults (20-45 years old). Half were of low socioeconomic status and the rest of medium-high status. The data were analyzed with SPSS-PC and EPI-INFO 6.0. Logistic regression and chi 2 were used for the analysis. Despite some differences found between age and socioeconomic status in regard to the characteristics desired for the idealized method, most of the participants expressed the same preferences. The results indicate that women would like the idealized method to be a cream, rather than a suppository, with no odor or flavor, to be colorless, to be placed in the vagina with an applicator well before coitus, and to offer protection against sexually transmitted diseases including AIDS. PMID- 9866007 TI - Women's preferences for vaginal antimicrobial contraceptives. III. Choice of a formulation, applicator, and packaging. AB - Novel vaginal formulations are under development to combat the increasing incidence of sexually transmitted diseases, including AIDS, and also unplanned pregnancies. A study was performed to determine women's preferences for different dosage forms (gel, cream, ovule/suppository, film, foam, tablet), width, length, and color of an applicator, and various types of packages. The study was conducted in Campinas, Brazil. A total of 635 women were interviewed, including both adolescents and adults and low and middle-high socioeconomic groups. The large majority of the women preferred a gel over a cream; both were preferred over the other methods. When asked which method they would not use, the film was most frequently identified, followed by the tablet and ovule. The primary reasons for selecting a particular dosage form were ease of use, absence of odor or the presence of a pleasant one, absence of color, and insertion with an applicator. The major reasons for not using a method were discomfort, "plastic" appearance, distrust of effectiveness, difficulty with insertion, messiness, and rigidity/hardness. The majority of the women liked the applicator shown. The prefilled single dose applicator was by far the preferred packaging. This information should aid in the development of consumer-friendly, vaginal formulations. PMID- 9866008 TI - Women's preferences for vaginal antimicrobial contraceptives. IV. Attributes of a formulation that would protect from STD/AIDS. AB - Vaginal formulations may have "dual" protective activity, against sexually transmitted diseases/AIDS and unplanned pregnancy. The attributes that women find acceptable or unacceptable for such dual protective methods were investigated. More than 50% of the women would not accept messiness, but it was more accepted for dual protective methods than for contraceptives. Very few women would use a dual protective method if it caused vaginal irritation, itching, swelling, or burning, problems associated with presently marketed methods. More than half of the women would use it if it appeared on the penis of their partner or required refrigeration. Use of an applicator to insert the formulation was generally preferred over a manual method. Most women preferred the formulation to be colorless or white, about 16% liked light colors, and about 10% liked darker colors. Almost half of the women were willing to pay up to $5.00 per application of a dual protective formulation, about 15% $3.00, and 30% $1.00. Dual protective methods seem highly acceptable and women would pay much more for them than for condoms. However, these methods should be free of problems usually associated with presently marketed formulations. PMID- 9866009 TI - Methadone in the management of narcotic addiction in pregnancy. PMID- 9866010 TI - Twin-twin transfusion syndrome: the challenge of etiology-based management decisions. AB - Studies published in the past year have attempted to model the pathophysiology of the twin-twin transfusion syndrome and also to establish the minimal diagnostic criteria for the condition. Many papers focused on treatment regimens and their sequelae. Aggressive treatment with serial amnioreduction continues to predominate, and appears to be more successful than conservative medical management, whereas selective feticide is reserved for cases that fail to respond to amnioreduction. Small series utilizing laser ablation and amniotic septostomy appeared; with laser proponents suggesting that their approach may carry a lower risk of neurological sequelae. Progress in ultrasound and Doppler techniques has begun to yield earlier and better diagnostic approaches which, coupled with an awareness of the possible anastomotic configurations, may allow for the selection of the management protocol most likely to succeed in each individual case. PMID- 9866011 TI - High risk pregnancy outcome by route of delivery. AB - This review examines the preferred route of delivery in accordance with neonatal and maternal outcome of three high risk pregnancy conditions: multiple pregnancy; delivery after cesarean section; and delivery of infants with macrosomia to mothers with gestational diabetes mellitus. The most common feature of all these conditions is the lack of information, based on large prospective controlled studies, available to the treating physician for choosing the delivery route of choice and for minimizing morbidity and mortality of both infant and mother. PMID- 9866012 TI - The maternal immune system in health and disease. AB - Recent studies have clarified some characteristics of trophoblast and immune cells in the decidua and peripheral blood during pregnancy and the roles they play in the maternal immune system, and have added further evidence to the theory that maternal T helper cell type 1/2 cytokine balance shifts towards T helper cell type 2 dominance during pregnancy. PMID- 9866013 TI - Shoulder dystocia. AB - Shoulder dystocia continues to represent a largely unpredictable and potentially disastrous obstetric emergency. Recent attention has been focused on the effectiveness of obstetric maneuvers employed to alleviate shoulder dystocia. Reports have also questioned the traditional thinking that brachial plexus injury is caused by application of excessive lateral traction to the fetal head. Rather, in-utero forces may underlie a significant portion of these injuries. PMID- 9866014 TI - Forced cesareans. AB - This review attempts primarily to provide guidance to physicians and others who may face the dilemma raised by a patient who refuses a recommended cesarean. This is, therefore, not a traditional literature review. The ethical-legal nature of the topic necessitates a different approach. Accordingly, this review briefly describes two recent cases in which courts have reversed orders to override a woman's refusal of consent, suggests different ways in which medical practice trends and emerging medical data may be implicated in forced cesarean cases, and then considers the implications of recent cases in the light of social-political factors. PMID- 9866015 TI - Unexplained elevated maternal serum alpha-fetoprotein: what is the appropriate follow-up? AB - Unexplained elevations of maternal serum alpha-fetoprotein exist in approximately 1% of the obstetric population. A consensus has been reached that these women face an increased risk of adverse pregnancy outcome. Whether their overall risk can be altered by the currently available surveillance modalities, however, remains controversial. Current research has focused on identifying those pregnancies with the highest risks of either fetal growth restriction, pre eclampsia, preterm delivery or intrauterine fetal demise. Markedly increased maternal serum alpha-fetoprotein (over 4.0 multiples of the median), elevations of other serum markers such as human chorionic gonadotropin and abnormal umbilical Doppler flow are associated with the greatest risk of poor pregnancy outcome. When initiating surveillance of the pregnancy with unexplained elevated maternal serum alpha-fetoprotein consideration of these factors is receiving increased attention. PMID- 9866016 TI - Postpartum hemorrhage and arterial embolization. AB - Postpartum hemorrhage remains a major cause of morbidity and mortality for the obstetric patient. A timely, stepwise approach to management can reduce the negative impact of this complication. Improvements in pharmacotherapy and surgical techniques have also improved outcome. Uterine artery embolization is an especially promising approach to the management of severe or refractory obstetric hemorrhage which is generally underutilized but has several advantages over other more traditional techniques. PMID- 9866017 TI - Fetal pulse oximetry. AB - Within the last ten years several groups adapted pulse oximetry to be used in the fetus. The obvious advantage of this technology is the fact that a biochemical parameter--the arterial oxygen saturation--can be measured continuously during delivery. Nevertheless, the continuous information about the fetal oxygenation during delivery has a couple of obstacles to surmount. It is well known that fetal reflectance pulse oximetry may be influenced by a number of artifacts. In addition, severe physiological considerations should remind us of the limited diagnostic value of saturation monitoring alone in order to predict fetal acidosis. Some recent articles deal with the predictive value of fetal pulse oximetry for fetal compromise. While it appears that the fetal wellbeing is more likely to be in accordance with a normal saturation measured by current pulse oximetry systems, the number of fetuses detected by pulse oximetry suffering from hypoxia seems to be low. Different authors describe a poor sensitivity to predict fetal compromise. One reason therefore may be the reduced precision of the oxysensor in the low saturation range. Therefore, we conclude that the current generation of fetal pulse oximetry sensors is not improving the quality of combined fetal monitoring of fetal heart rate and fetal scalp blood analysis. PMID- 9866018 TI - General obstetrics. PMID- 9866019 TI - [A model of chronic situational stress and its effect on the rate of aging and the longevity of rats]. AB - In rat experiments a new model of situational chronic stress, with the use of multicomponent natural stimulating influences, has been elaborated. The study revealed two subpopulations with longer and shorter lifespan in the group of stressed rats vs. control group. The survival of stressed animals was correspondent with the changes of adaptive indices. Thus the situational chronic stress has the geropromotive or geroprotective effect which depends on the organism's adaptive capacities. PMID- 9866020 TI - [The electrical reactions of endothelial cells in situ to ATP action]. AB - The changes in the membrane potential (MP) of in situ endothelial cells from guinea pig aorta was studied using patch-champ technique under ATP stimulation. Extracellular ATP is shown to evoke the complex changes in endothelial MP: initial short-lived depolarization and subsequent maintained hyperpolarization. Extracellular calcium buffering as well as addition of extracellular Ni2+ made the hyperpolarization shorter. After the buffering of intracellular calcium ATP addition evoked only depolarization which was not observed in the absence of extracellular sodium. The hyperpolarization under ATP stimulation was absent after emptying of intracellular calcium stores. It was concluded that hyperpolarization in response to ATP is initiated by calcium release from intracellular stores followed by activation of calcium-dependent potassium channels, the maintained phase of hyperpolarization is provided by extracellular calcium entry and initial depolarization by extracellular sodium entry into EC. PMID- 9866022 TI - [The modulating action of serotonin on single calcium channels in the neurons of the snail Helix pomatia]. AB - The influence of a neurotransmitter serotonin (5-hydroxytriptamine, 5-HT) on single Ca2+ channel activity was studied on unidentified neurons of the snail Helix pomatia. Our findings showed that 5-HT influences the channel molecule in indirect manner, as it appeared effective at its application by adding to the bath solution, that had no contact with the tested membrane fragment. This finding excludes the possibility of direct binding of 5-HT to the channel molecule and shows that the effect is really mediated through a cytoplasmic messenger, cAMP-dependent cascade of phosphorylation. Also our investigations shows, that only one type of Ca2+ channel with the unitary conductance of 5 pS was identified in this kind of neurons. PMID- 9866021 TI - [The effect of chloditan on synthetic processes in the adrenal gland cells of newborn piglets]. AB - In experiments with a primary culture of adrenal cells, it has been studied the influence of chloditane, as a specific inhibitor of adrenocortical function, on the synthetic processes in the newborn pig adrenal cells, whose organ and cell cultures are more and more used as transplantation material for the treatment of adrenal insufficiency. It has been shown that incubation of adrenocorticocytes with chloditane (final concentration in culture medium: 1 to 10,000 ng/ml) during 24 hours had a marked negative dose-dependent effect on the indices of inclusion of 3H-thymidine and 3H-uridine into the nucleic acids. In the presence of a maximum chloditane concentration, inclusion made, respectively, 16.9 and 46.0% from control. A trustworthy effect of chloditane on the indices of 3H-leucine inclusion (protein synthesis) has been established only for concentration 10 to 100 ng/ml and made near 70% from control. It has been discussed a possible mechanism of chloditane action on the synthetic processes in adrenal cells. PMID- 9866023 TI - [The combined effect of work stress, fatigue and circadian rhythms on the efficiency of mental activity in operators]. AB - Using the multiple psychophysiological testing for 73 Chornobyl NPP control room operators (N = 1270 subject tests) there were revealed, that the information processing effectiveness significantly decreases during night shift, whereas during morning and evening shifts it is maintaining at the constant level practically. This is caused, on the one hand, by the circadian rhythms peculiarities of the different occupationally vital performance parameters (performance speed functions deteriorate under the absence of the improvement of others at night, while at the day time there are observed the different directioned changes for the different functions), on the other hand--by the more sensitivity of the body to the fatigue and work tension at night. The significant expression of the manifestation of natural circadian rhythms in the variations of the professionally important parameters of the operators' mental activity has been connected with the increased influence of the unfavourable factors on the workers' functional body state. There are revealed the more sensitivity of the information processing speed parameters to the fatigue in comparison with an accuracy, and the more sensitivity of the mental activity accuracy to the lengthy supported level of the nervous-emotional work tension in comparison with a speed. PMID- 9866024 TI - [The effect of linoleyl hydroxamic acid on lipid peroxidation processes and on the enzymatic activity of the antioxidant system in rats under hypoxia]. AB - It was studied the effect of new blocker of lipoxygenase way of arachidontic acid metabolism--linoleyl-hydroxamic acid--on the status of lipid peroxidation and antioxidant system in blood, heart, liver, lung and brain in rats under acute hypoxic hypoxia. It was shows that preliminary introduction of this substance lead to increase of antioxidant enzymes activity--superoxide dismutase and ceruloplasmine, and to decrease of lipid peroxidation level during hypoxia in blood, heart, liver and lung (without brain). PMID- 9866025 TI - [The mechanisms of the lymph-stimulating action of liposomes]. AB - The mechanisms of lymph-stimulating action of different liposomes (lecithin cholesterin, lecithin) were studied in the experiments on dogs. It has been shown an absence of interaction between changes in the lymph outflow rate and system circulation parameters when injecting any liposomes. Moreover, water and protein filtration from the vascular bed into the interstitial space was lowered due to liposomal injection. But oxygen tension in the muscular tissues remained on the level closed to the normal one. In contrast to the intravenous injection, the subcutaneous administration of liposomes has induced the stimulation of lymph outflow, which was more expressed on both volume (nearly by 40%) and duration (nearly by 50%). The experiments with limited and crossed blood flow have demonstrated a regional character of liposomal lymph-stimulating. The mechanisms of such liposomal action may be related to both direct action of liposomes on the lymphatic smooth muscle contractility in the lymphatic vessels and the increased lymph production in the site of liposomal injection due to free radical oxidation processes being limited in that area. The micellar form of phospholipids (essential) has demonstrated no lymph-stimulating action. PMID- 9866026 TI - [The structural organization of the aortic endothelium in hypercholesterolemia and the effect of L-arginine]. AB - Electron microscopic investigation of the rabbit [correction of rat] thoracic aorta revealed endothelium to be most vulnerable to the damage in experimental hypercholesterolemia. Administration of precursor of nitric oxide L-arginine against the background of hypercholesterolemia resulted in activation of functional activity of the endothelium of aorta. This notion is supported by an increase of the quantity of protein-synthesizing structures (ribosomes, polysomes, canaliculi of endoplasmic network), secretory granules vs rabbits on atherogenic diet. At stimulation of NO-synthase activity of endothelium the dilatory reactions of aortal strips to acetylcholine are improved. PMID- 9866027 TI - [The antioxidant action of taurine in acute hypoxic hypoxia]. AB - It was found that preliminary treatment by amino acid taurine protected rats from lipid peroxidation intensification (expressed in terms of malondialdehyde and conjugated dienes contents) in the liver, brain and heart under acute severe normobaric hypoxic hypoxia. The mechanisms of the antioxidant action of taurine are connected to the prevention of lactate accumulation in tissues and cell membrane structure disorders (expressed in a decrease of membrane Na+, K(+) ATPase activity). It was also shown that taurine reduced significantly a decrease of glutathione antioxidant system activity protecting tissues against reduced glutathione pool depletion and preventing a decrease of glutathione reductase and glutathione peroxidase activities in acute severe hypoxia. PMID- 9866028 TI - [The effect of dexamethasone on the reactions of the blood system in inflammation]. AB - On the model of carrageenan-induced acute aseptic peritonitis in rats it is shown that under influence of dexamethasone granulomonocytopoiesis, efflux of leukocytes to blood and their accumulation in inflammatory focus are increased and earlier completion of leukocytic reaction is observed and that the antiinflammatory effect of dexamethasone is mainly realized by the way of increasing of defence-adaptative blood system's reactions. PMID- 9866030 TI - [The dependence of thrombocyte aggregation on temperature]. AB - The effect of temperature (range of 4-44 degrees C) upon induced aggregation of platelets from normal subjects was studied. It was shown that temperature dependences of aggregation degree and rate have maxima. A relationship between aggregation agonist kind and some features of the temperature dependence curve of the aggregation parameters is established. It was shown that the limitation of platelet membrane lipids mobility by means of lipids peroxidation leads to the shift of maxima of the temperature dependence curve of platelet aggregation degree and rate. It was concluded that character of aggregation temperature dependence was governed by phase transition in platelet membrane lipids at room temperature. PMID- 9866029 TI - [The effect of insulin on the function of the lysosomal apparatus of the neutrophilic leukocytes during the formation of a stress syndrome]. AB - In experiments on male rabbits with the lack of insulin it is been revealed violations after the past immobilization the intensivity and duration of neutrophilic leukocytosis decrease, contents of lysosomes in neutrophils, activity of acid phosphatase. By lysosomal ferments don't determine the can observer discordance of processes of coagulation, fibrinolysis, kininogenesis. PMID- 9866031 TI - [The toxic component in the development of lung damages in uremia]. AB - Renal ischemia causes accumulation of middle-sized molecular components of peptide and non-peptide character in the blood. These substances, obtained from the blood by method of ultrafiltration are able to cause the marked lung injuries with formation of leukocyte and erythrocyte aggregations, endothelial lung cell injured and with the phenomenon of hyperhydration. The indicated changes are proportional by their expressiveness to the dose of administrated middle-sized molecules. The conclusion can be made that toxic component, associated with accumulation of biologically active middle-sized molecules in the blood plays a significant role in the development of nephrogenic pulmonary edema. PMID- 9866032 TI - [Spasmophilia in children and the anticonvulsant properties of a new Ukrainian preparation containing a pyrimidine derivative]. AB - Tablets valcophen, which are containing the pyrimidine's derivative, have an acute anticonvulsive activity. Tablets valcophen and granules phenobarbital for children are having the same activity, which have been marked about lower intensity convulsions, quantity of a lived animals and also lengthening a time of deathing mice. These investigations are showing that tablets valcophen may be used for treatment convulsive state in children. PMID- 9866033 TI - [The modelling of local radiation injuries to the skin]. AB - The models of local radiation injuries of skin--radiation ulcer, radiation epidermitis, radiation dermatitis in guinea-pigs and rabbits are developed. Experimental reproduction local radiation injuries by different doses, regimes and spices of ionizing irradiation was carried out. Detailed description of radiation action on skin is given. Pathomorphologic characteristic of radiation ulcers is given. Models may be used for pathogenesis learning and experimental therapy of local skin radiation injures. PMID- 9866034 TI - [A method of preparing the results of biomedical research for statistical processing]. AB - The method of preparing experimental findings based on their rationing which proceeds their statistical processing has been supposed. It has been shown that the supposed approach allows to find out real changes in the average values which are not revealed without rationing the initial findings. PMID- 9866035 TI - [The role of oxygen radicals in the physiology and pathology of human sperm]. AB - Reactive oxygen species in low doses are necessary compound of sperm capacitation and hyperactivation. Superoxide anion, hydroxyl radical and hydrogen peroxide initiate sperm capacitation. The edding of antioxidant enzymes inhibits the spontaneous and induced sperm hyperactivation. The process of capacitation is accompanied with the superoxide anion production output by spermatozoa. High doses of reactive oxygen species block the sperm motility through the inhibition of ATP synthesis by the mitochondrial enzymes and cell membrane compounds injury. PMID- 9866036 TI - [The contribution to physiological science of the scientists of the O. O. Bohomolets Institute of Physiology of the National Academy of Sciences of Ukraine (the basic achievements)]. PMID- 9866037 TI - [Criteria for evaluating guidelines in medical care]. PMID- 9866038 TI - [Tinea of the hairless skin. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866039 TI - [Tinea unguium--onychomycosis. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866040 TI - [Candidiasis of the female genitals. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866041 TI - [Oral candidiasis. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866042 TI - [Candidiasis of the skin. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866043 TI - [Basal cell carcinoma (basalioma; basal cell epithelioma). Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866044 TI - [Squamous epithelial carcinoma of the skin including the lower lip and eyelids (spinalioma, spinous cell carcinoma). Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e.V]. PMID- 9866045 TI - [Cutaneous neuroendocrine carcinoma (Merkel cell carcinoma). Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866046 TI - [Malignant melanoma. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866047 TI - [Kaposi sarcoma. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866048 TI - [Dermatofibrosarcoma protuberans. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866049 TI - [Cutaneous lymphomas. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866050 TI - [Instrumental intermittent compression. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866051 TI - [Chronic venous insufficiency (CVI). Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866052 TI - [Venous diagnosis with light reflection rheography/photoplethysmography. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866053 TI - [Medical compression stockings. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866054 TI - [Medical stocking for prevention of thrombosis. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866055 TI - [Surgical management of venous diseases]. PMID- 9866056 TI - [Phlebologic compression bandage. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866057 TI - [Prevention of thromboembolism. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866058 TI - [Diagnosis and therapy of superficial thrombophlebitis. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866059 TI - [Deep venous thrombosis of the leg/pelvis. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866060 TI - [Ulcus cruris venosum. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866061 TI - [Pharmacotherapy in proctology. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866062 TI - [Anal eczema. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866063 TI - [Condylomata acuminata of the anorectal region. Quality Assurance Committee of the German Society of Dermatology and the Professional Organization of German Dermatologists e. V]. PMID- 9866064 TI - A matched study to determine whether low-dose aspirin without heparin improves pregnancy rates following frozen embryo transfer and/or affects endometrial sonographic parameters. AB - PURPOSE: The objective of the matched, controlled study was to determine whether low-dose aspirin therapy without heparin improves pregnancy rates following frozen embryo transfer. METHODS: Thirty-six women who did not achieve a pregnancy following fresh embryo transfer and who had frozen embryos available for another transfer were included. Eighteen women were treated with 81 mg aspirin from day 2 of the cycle through pregnancy testing. If the beta-human chorionic gonadotropin level was positive, aspirin was continued through the pregnancy. Eighteen women were not given aspirin. The mean outcome variables were pregnancy and implantation rates. RESULTS: The clinical pregnancy rate in the aspirin group was 11.1%, compared with 33.3% for the controls, and implantation rates were 2.9 and 10.9%, respectively. CONCLUSIONS: No positive effects of low-dose aspirin therapy on pregnancy rates following frozen embryo transfer were observed. PMID- 9866065 TI - Low plasma levels of hCG after 10,000-IU hCG injection do not reduce the number or maturation of oocytes recovered in patients undergoing assisted reproduction. AB - PURPOSE: Our purpose was to determine whether there is a correlation between human chorionic gonadotropin (hCG) blood levels and oocyte maturation. METHODS: Three samples of blood were obtained at different times from hCG administration as follows: 12 hr, 36 hr, during oocyte recovery, and at 84 hr, when the patient comes for embryo transfer. RESULTS: A total of 5036 oocytes was retrieved from 404 patients prospectively recruited between April 1996 and March 1997. The percentage of metaphase-II oocytes at different blood levels ranged from 84 to 88%. The general trend does not show any significant increase in percentage of metaphase-II oocytes in association with an increasing serum hCG concentration. CONCLUSIONS: The results of this study suggest that at 12, 36, and 84 hr after hCG administration, levels as low as 50, 45, and 9 IU/L of hCG, respectively, are equally potent as higher levels at initiating maximal oocyte maturity. PMID- 9866066 TI - Follicular fluid insulin-like growth factor-I and insulin-like growth factor binding protein-1 and -3 vary as a function of ovarian reserve and ovarian stimulation. AB - PURPOSE: Follicular fluid concentrations of insulin-like growth factor (IGF)-I, IGF-II, IGF-binding protein (BP)-1, and IGFBP-3 in 57 women undergoing in vitro fertilization and embryo transfer were examined to determine whether levels reflected differences in patients' exposure to gonadotropin stimulation and a diminished ovarian reserve. METHODS: Preovulatory follicular fluid was obtained from both gonadotropin-stimulated and unstimulated cycles. Subjects were grouped according to normal or decreased ovarian reserve and whether or not they received gonadotropin stimulation. RESULTS: The mean follicular fluid concentrations of IGF-I and IGFBP-1 were significantly lower in the "decreased" ovarian reserve group compared with the "normal" ovarian reserve group, with no change in estradiol or IGF-II levels. This resulted in a decreased molar IGF-I: BP ratio and an increased molar IGF-II:IGFBP-1 ratio. In unstimulated cycles, mean follicular fluid concentrations of IGFs did not differ significantly compared with those in stimulated cycles, whereas concentrations of IGFBP-1 and IGFBP-3 were significantly lower, leading to higher molar ratios of the IGFs to the binding proteins. CONCLUSIONS: Follicular fluid IGF and binding proteins vary as a function of ovarian reserve and gonadotropin stimulation. This may reflect either differences in oocyte quality or a suboptimal follicular fluid environment. PMID- 9866067 TI - Low blastocyst formation rates in day-2 fertilized oocytes. AB - PURPOSE: To examine the blastocyst formation rates of day-2 fertilized oocytes. METHODS: A retrospective study of the outcomes/blastocyst formation of day-2 fertilized oocytes was undertaken. RESULTS: Fertilization rates of day-1 and -2 oocytes by intra-cytoplasmic sperm injection were similar. The development frequencies to four cells were similar. However, the blastulation rates were significantly lower from the day-2 fertilized eggs. The fertilization rates from day-2 conventional in vitro fertilization reinsemination were lower than the fertilization rates of day-1 oocytes. The blastulation rates from day-2 fertilized eggs were also lower than the rates from day-1 fertilized eggs in the in vitro fertilization group. CONCLUSIONS: Fertilization is not a good indicator to predict the viability of fertilized oocytes. Day-2 fertilized oocytes had significantly lower blastocyst formation rates than the rates from day-1 fertilized oocytes. PMID- 9866069 TI - Effects of reducing insemination time in human in vitro fertilization and embryo development by using sibling oocytes. AB - PURPOSE: Recent studies showed a beneficial effect of reducing the time of sperm oocyte interaction on fertilization, division, and implantation rates of the oocytes obtained from randomized patients. In the present study, the effects of reduced insemination time on fertilization and embryo development were evaluated by using sibling oocytes from the same patient. METHODS: A total of 464 oocytes from 36 patients was randomly allocated to be inseminated for either 1 hr (reduced) or 18 hr (regular). RESULTS: Fertilization rates were not significantly different between reduced (135/229; 59%) and regular (150/235; 64%) groups. Cleavage rates and embryo quality were similar in both groups. A total of 135 embryos (73 from the reduced and 62 from the regular group) was transferred to 36 patients. Thirty-four embryos implanted in 18 patients (25.2% implantation and 50.0% pregnancy rates). CONCLUSIONS: Fertilization, cleavage, and embryo development from 1-hr insemination is comparable, not superior, to those from an 18-hr insemination time, which is commonly used in in vitro fertilization programs. These data suggest that reduced insemination time can be used during in vitro fertilization to avoid unnecessarily longer exposure to spermatozoa. PMID- 9866068 TI - Pretreatment with an oral contraceptive is effective in reducing the incidence of functional ovarian cyst formation during pituitary suppression by gonadotropin releasing hormone analogues. AB - PURPOSE: Our purpose was to assess the effect of pretreatment with oral contraceptives (OCs) on the formation of functional ovarian cysts during pituitary suppression with gonadotropin-releasing hormone (GnRH) agonists, subsequent follicular development, and pregnancy rates. METHODS: A retrospective case-controlled study of 31 in vitro fertilization (IVF) patients, all of whom in a previous cycle had commenced the long protocol of GnRH-agonist (Buserelin) in the early follicular phase and were pretreated in a subsequent cycle with 2 weeks of an OC containing 30 micrograms of ethinyl estradiol and 150 micrograms of desogestrel prior to GnRH-agonist administration, was undertaken. Follow-up visits were arranged after a minimum of 11 days of GnRH-agonist administration and weekly thereafter until pituitary suppression was achieved. RESULTS: Cysts were detected in 16 (51.6%) of the 31 patients not pretreated with OCs, and in 0 (0%) of the 31 patients pretreated with OCs (odds ratio = 67.1; 95% confidence interval = 5.6-350.7). Patients pretreated with OCs achieved pituitary suppression more rapidly (median difference = 4 days; 95% confidence interval = 2 7) and had comparable gonadotropin requirements and pregnancy rates. CONCLUSIONS: Pretreatment with OCs prior to pituitary suppression in the early follicular phase decreases ovarian cyst formation, without an apparent effect on subsequent follicular recruitment or pregnancy rates. PMID- 9866070 TI - Centrifugal pulsing increases the recovery of viable spermatozoa. AB - PURPOSE: Our purpose was to compare the number and quality of spermatozoa recover following the standard centrifugation method with those after pulsing the relative centrifugal field during centrifugation. METHODS: A prospectively controlled in vitro study using ejaculates from 10 healthy sperm donors was undertaken. Number of sperm and sperm motility at 0, 3, and 24 hr post-incubation at 5 degrees C in TEST-yolk results determined the efficacy of the centrifugation techniques. RESULTS: The number of sperm and sperm motility at 0, 3, and 24 hr post-incubation at 5 degrees C in TEST-yolk were significantly higher for the sperm recovered following pulse centrifugation compared with those after standard centrifugation. CONCLUSIONS: Pulsing the relative centrifugal field during centrifugation yields a higher number of viable spermatozoa. PMID- 9866071 TI - Evaluation of endometrial inflammation by quantification of macrophages, T lymphocytes, and interleukin-1 and -6 in human endometrium. AB - PURPOSE: Our purpose was to determine the impact of endometrial inflammation on fertility. METHODS: The study population included 49 women: 5 women bearing an intrauterine device, 33 sterile patients, and 11 fertile women. Endometrial biopsies were performed between day 17 and day 21 of the cycle. Immunostaining was performed with an ABC kit. Monoclonal antibodies were CD68 for macrophages, CD3 for T lymphocytes, and B-E8 for interleukin (IL)-6; IL-1 alpha and IL-1 beta were stained with polyclonal antibodies. A double immunofluorescence staining was performed to identify endometrial cells. Macrophage and lymphocyte rates were defined as the percentage of stained cells by field. Double staining was compared among macrophages, T lymphocytes, and anti-IL antibodies in each group. RESULTS: The macrophage rate was high for women bearing an intrauterine device, intermediate for sterile patients, and low for fertile women (P = 0.03). There was a lack of double staining between activated T lymphocytes and anti-IL-6 antibody in tubal sterility and endometriosis. Endometrial macrophages were stained with anti-IL antibodies for all groups. CONCLUSIONS: Endometrial macrophages seem to be an interesting parameter for evaluating endometrial inflammation. In tubal infertility and endometriosis, the lack of expression of IL-6 in T lymphocytes could participate in impaired embryo implantation. PMID- 9866072 TI - Isolation and purification of an early pregnancy factor-like molecule from culture supernatants obtained from lymphocytes of pregnant women: II. Identification of the molecule as a Fc-receptor-like molecule: a preliminary report. AB - PURPOSE: Early pregnancy factor (EPF)-like activity from culture supernatants obtained from stimulated lymphocytes of pregnant women was characterized and identified. METHODS: The enzyme-linked immunosorbent assay depending on the presence of "Fc" receptors on bovine spermatozoa was used to identify the EPF like molecule purified by gel filtration and reverse-phase high-performance liquid chromatography. RESULTS: The results indicated that the crude lymphocyte culture supernatant, the EPF-positive G IV fraction obtained on gel filtration, and the EPF-positive reverse-phase high-performance liquid chromatography protein readily bound with the different concentrations of aggregated human gamma globulin in a manner similar to that in which the standard control of aggregated human gamma-globulin binds to the bovine spermatozoa. CONCLUSIONS: EPF-like activity synthesized and secreted by lymphocytes during pregnancy may be a Fc receptor-like molecule. PMID- 9866073 TI - A successful pregnancy with in vitro fertilization and embryo transfer in an infertile woman with Kartagener's syndrome: a case report. PMID- 9866074 TI - Negative life events and the adjustment of school-age children: testing protective models. AB - Investigated the association between negative life events and protective factors in predicting the adaptive, emotional, and behavioral functioning of school-age children. Three possible models of this relation were tested using hierarchical analyses: the compensatory model, the challenge model, and the immunity/vulnerability model. Participants were 140 children between the ages of 8 to 13.6 years. Inconsistent with all 3 models, negative life events were not associated with adaptive or internalizing behavior. However, consistent with the compensatory model, both negative life events and protective factors contributed independently to the prediction of externalizing behavior. Also, a significant interaction was found in predicting internalizing behavior for the female-only sample showing girls with significant negative life events and social support demonstrating less internalizing behavior. PMID- 9866075 TI - Reactive aggression among maltreated children: the contributions of attention and emotion dysregulation. AB - Examined the complex interplay among emotion, attention, and aggression in a sample of 141 maltreated and 87 non-maltreated impoverished, inner-city children. Data were collected during a summer day camp, which provided an ecologically valid setting for studying children's behavior in social contexts. Maltreated children were more likely than non-maltreated children to be aggressive, with findings suggesting that physically abused children were at heightened risk for reactive aggression. Maltreated children also evidenced attention deficits, and subclinical or nonpathological dissociation was more likely among children who had experienced physical or sexual abuse. A history of abuse also predicted emotion dysregulation, affective lability/negativity, and socially inappropriate emotion expressions. This emotion dysregulation, fostered by poor attention modulation, was a mechanism of the effects of maltreatment on reactive aggression. PMID- 9866077 TI - Factors influencing elementary school teachers' ratings of ADHD and ODD behaviors. AB - Examined factors that influence teachers' ratings of children with either attention deficit hyperactivity disorder (ADHD) or oppositional defiant disorder (ODD). 105 teachers watched 2 videotapes--1 depicting a normal child and the other a child with either ADHD or ODD--and rated each child using 2 different questionnaires. Results indicated that teachers accurately rated the child on the ADHD versus ODD tape as having significantly more inattention and hyperactivity but significantly less oppositionality. However, effect sizes indicated the presence of a unidirectional, negative halo effect of oppositional behaviors on ratings of hyperactivity and inattention. Teachers appeared less biased in their judgments when using a well-operationalized rating scale. Finally, knowledge, education, and experience with children with ADHD generally had no effect on the accuracy of teachers' ratings. PMID- 9866078 TI - Smokeless tobacco and cigarettes: differential attitudes and behavioral intentions of young adolescents toward a hypothetical new peer. AB - Examined adolescents' perceptions of cigarette smokers and smokeless tobacco users. Participants were 562 middle school students in rural Florida who viewed 1 of 6 videotapes of a hypothetical peer (i.e., actor) who would soon be attending their school. The videotapes differed only as a function of sex (boy or girl) and tobacco condition (no tobacco, cigarette, smokeless tobacco). After viewing the videotape, participants completed 2 measures designed to assess attitudes and behavioral intentions toward the peer. Results indicated that (a) the actor in the no-tobacco condition was rated more favorably than actors in the other two conditions, although the actor in the smokeless-tobacco condition was rated more favorably than the actor in the cigarette condition; (b) girls viewed the actor in smokeless-tobacco condition more favorably than did boys; (c) compared to nonsmokers, adolescents with a cigarette use history provided more favorable ratings for the actor in the cigarette condition. Taken together, results suggest that different types of tobacco use may have a different impact on social image within the young adolescent population. PMID- 9866076 TI - Criterion validity and the utility of reactive and proactive aggression: comparisons to attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder, and other measures of functioning. AB - Examined the criterion validity and the utility of Dodge and Coie's (1987) measure of reactive and proactive aggression. Participants were 405 children in kindergarten through 5th grade attending an urban elementary school. Examined criterion validity by testing whether reactive aggression or proactive aggression was significantly correlated with criterion measures of overall impairment as measured by the Impairment Rating Scale (Pelham, Gnagy, et al., 1996), classroom behavior as measured by a frequency count of classroom rule violations, and peer adjustment as measured by teacher ratings of peer behavior on the Pittsburgh Modified IOWA Conners (Pelham, Milich, Murphy, & Murphy, 1989). Examined utility by testing whether reactive aggression or proactive aggression was significantly correlated with criterion measures after controlling for each other and after controlling for attention deficit hyperactivity disorder, oppositional defiant disorder, and conduct disorder as measured by the Disruptive Behavior Disorder Rating Scales (Pelham, Gnagy, Greenslade, & Milich, 1992). Results showed good evidence of criterion validity for both reactive and proactive aggression, but mixed evidence for their utility. PMID- 9866079 TI - Changing self-esteem in children and adolescents: a meta-analytic review. AB - Conducted a meta-analytic review of 116 studies, which indicated significant improvement in children's and adolescents' self-esteem and self-concept, and significant concomitant changes in behavioral, personality, and academic functioning. Interventions specifically focused on changing self-esteem and self concept were significantly more effective (mean effect size = 0.57) than programs focused on another target, such as behavior or social skills (0.10). Treatment programs were also more effective (0.47) than primary prevention programs (0.09) in changing self-esteem. Four variables emerged as significant predictors of self esteem outcomes: 2 methodological features (type of design and control group), the use of a theoretical or empirical rationale, and the type of program (treatment or prevention). Future research needs to examine the causal connection between changes occurring in self-esteem and other areas of adjustment, assess intervention success for different ethnic groups and for children of different ages and sex, and determine the long-term impact of interventions. PMID- 9866080 TI - Psychological symptomatology following parental death in a predominantly minority sample of children and adolescents. AB - Examined the psychological correlates related to experiencing the death of a parent, the main and interactive effects of sex, race, and age on youth distress and the degree of cross-informant correspondence on the outcome measures. The predominately minority sample included 80 bereaved youth and 45 nonbereaved youth. Youth and their legal guardians completed a battery of questionnaires, including measures assessing the youth's psychological symptomatology. Results revealed that bereaved youth manifested greater psychological and behavior problems than their nonbereaved counterparts on guardian-reported measures (Child Behavior Checklist [CBCL]). The clinical significance of parental death experienced during childhood is indicated by the magnitude of distress exhibited by the bereaved sample; almost one quarter of bereaved youth scored in the clinical distress range (T score > or = 63) on the CBCL Externalizing and Internalizing Distress scales. The effect of parental death on guardian-reported externalizing distress was moderated by race, such that distress levels did not significantly differ between bereaved and nonbereaved minority youth but did differ significantly among bereaved and nonbereaved nonminority youth. Finally, the degree of cross-informant agreement was relatively low but consistent with prior studies. Study implications for interventions with bereaved youth and directions for future research are discussed. PMID- 9866081 TI - Mother- and father-rated competence, child-perceived competence, and cognitive distortions: unique relations with children's depressive symptoms. AB - Examined the unique relations of mother- and father-rated social and academic competence, children's perceived competence, and cognitive distortions to children's depressive symptoms. Participants were 389 8- to 12-year-old children and their mothers and fathers. Mother- and father-rated competence did not explain variability in child-reported depressive symptoms beyond that explained by child-perceived competence. Children's perceived competence was not related to mothers' ratings of children's depressive symptoms after statistically adjusting for mother-rated competence. Although child-perceived academic competence was not related to fathers' ratings of children's depressive symptoms after adjusting for father-rated academic competence, child-perceived social competence explained significant variability in fathers' reports of depressive symptoms, beyond that explained by father-rated social competence. Children's cognitive distortions were significantly related to both child- and mother-reported depressive symptoms after adjusting for the relations of both parent-rated competence and children's perceived competence. The results are discussed in light of competency-based and cognitive models of depression and highlight the need to consider the interplay of parent-rated competence, child-perceived competence, and cognitive distortions in examining childhood depression. PMID- 9866082 TI - Psychometric evaluation of the social phobia and anxiety inventory for children: concurrent validity and normative data. AB - Examined the relation of the Social Phobia and Anxiety Inventory for Children (SPAI-C) to the Social Anxiety Scale for Children-Revised (SASC-R). The association between the SPAI-C and the SASC-R was moderate, suggesting that the measures assess overlapping, although not identical, constructs. Initial normative data are reported on the SPAI-C for a community sample of 277 4th through 6th-grade children. Girls reported higher social anxiety on both measures than did boys. Notably, a substantial proportion of children were found to exceed suggested cut-off scores on both measures for identifying children with high levels of social anxiety, which suggests the importance of screening elementary age children for social fears. PMID- 9866083 TI - Evaluation of cognitive-behavioral group treatments for childhood anxiety disorders. AB - Undertook an evaluation of a cognitive-behavioral group family-based intervention for childhood anxiety disorders in Brisbane, Australia. The treatment aimed to provide children and their families with skills in the management of anxiety and avoidance, problem solving, and mutual family support. Children (n = 60) ranging from 7 to 14 years old who fulfilled diagnostic criteria for separation anxiety, overanxious disorder, or social phobia were randomly allocated to 3 treatment conditions: group cognitive-behavioral therapy (GROUP-CBT), group cognitive behavioral therapy plus family management (GROUP-FAM), and wait list (WL). The effectiveness of the interventions was evaluated at posttreatment and 12-month follow-up. Results indicated that across treatment conditions, 64.8% of children no longer fulfilled diagnostic criteria for an anxiety disorder in comparison with 25.2% of children on the wait list. At 12-month FU, 64.5% of children in the GROUP-CBT group and 84.8% of children in the GROUP-FAM group were diagnosis free. Comparisons of children receiving GROUP-CBT with those receiving GROUP-FAM on self-report measures and clinician ratings indicated marginal added benefits from GROUP-FAM treatment. Results show that CBT interventions for childhood anxiety disorders can be effectively administered in a group format. PMID- 9866084 TI - Play, creativity, and adaptive functioning: implications for play interventions. AB - Discussed the importance of play in creative problem solving and its implications for play interventions. Theory and research in the areas of play and creative cognitive processes and play and creative affective processes are reviewed. Play has been associated with the development of creative problem solving. Creative problem solving is thought to be a resource for everyday coping and adjustment. Play intervention studies that investigate the effects of specific aspects of play on specific criteria would be appropriate for the creative problem-solving and coping area. A systematic program integrating laboratory research on play and creativity, research on play techniques, and play intervention with specific populations is needed. PMID- 9866085 TI - Temperament of preterm infants at 4 months of age: maternal ratings and perceptions. AB - The purpose of this study was to compare temperament of preterm infants born at 34 weeks gestation or earlier with that of healthy, term infants at 4 months of age by maternal ratings and maternal perceptions and to examine the role of social support on maternal assessments of infant temperament. The mothers of preterm infants completed the Early Infancy Temperament Questionnaire (EITQ) and the Maternal Social Support Index when the child reached 4 months adjusted age. The EITQ assesses infant temperament by maternal ratings of specific infant behaviors and by mothers' global perceptions of infant temperament. Mothers of healthy term infants completed the same instruments when they brought their 4 month-old infant for a pediatric visit. Mothers rated preterm infants as having more negative mood (p = .01) and being less adaptable (p = .03) than term infants. However, mothers perceived that preterm infants were more difficult overall (p = .00001), had more negative mood (p < .005), were less regular (p = .02), and were less adaptable (p = .03). Mothers perceive preterm infant temperament to be more difficult than that of term infants and to be more difficult than is indicated by ratings of individual infant behaviors. PMID- 9866086 TI - Elementary school-age children's developmental understanding of the causes of cancer. AB - This study examines children's conceptual understanding and factual knowledge of the causes of cancer. Using a standardized, developmentally based, semistructured interview (ASK [AIDS (acquired immunodeficiency syndrome) Survey for Kids]), 784 children (43% black, 38% white, and 18% Hispanic; 48% female) in kindergarten through sixth grade attending six public elementary/middle schools in New Haven, Connecticut, were asked open-ended questions about the causes of cancer and, for comparison, the causes of colds and AIDS. Responses were scored for level of conceptual understanding and coded for factual content and factual accuracy. The level of conceptual understanding for causality of cancer increased consistently as grade level increased. When comparisons were made among the illnesses, children's level of conceptual understanding was significantly lower for the causes of cancer than for the causes of colds (p < .0001), but not significantly different from that of AIDS. Although the single most frequent cause of cancer mentioned was cigarettes/smoking (24%), more than one in five students stated that casual contact or contagion was a cause of cancer. More children cited causal contact/contagion than cited the following factually accurate or logically contributory causes combined: poor diet, air/water pollution or overexposure to sun, alcohol, and old age. Slightly more than one half of students in kindergarten through sixth grade worried about getting cancer, and the vast majority (80%) knew that cancer could be fatal. Children have a less sophisticated conceptual understanding of cancer than of colds and a very limited factual knowledge base for cancer, and thus they have the capacity to increase both their understanding and knowledge. These results have implications for the creation of developmentally appropriate cancer prevention curricula for elementary school-age children. PMID- 9866087 TI - Influence of birth weight on educational outcomes at age 9: the Miami site of the Infant Health and Development Program. AB - The Miami site of the Infant Health and Development Program, an early intervention for infants born low birth weight (LBW) and preterm, was investigated. Analyses unique to this sample were required because it was the only site that selected a normal birth weight (NBW) comparison group and had the lowest sociodemographic characteristics. Epidemiological methods determined the effects of LBW and early intervention on school outcomes. Children born LBW who did not receive intervention had an increased risk of scoring below the 10th percentile on achievement tests and were placed in special education three times more often than their peers born NBW. The school outcomes of children born LBW who received intervention were consistently better than those who did not, but were worse than children born NBW; however, differences did not reach statistical significance. Children born LBW who did not receive intervention are at significant risk for poor school outcomes compared with their peers born NBW. PMID- 9866088 TI - Early detection of social interaction problems: development of a social interaction instrument in young children. AB - Children with developmental or psychiatric disorders often have problems with social interaction. This study reports on the development of the Ghuman-Folstein Screen for Social Interaction (SSI), a parent/caregiver questionnaire designed to measure the capacity for basic social interaction skills across a variety of contexts in preschool children. The SSI was administered to 51 clinically referred children with a high probability of deficits in social interaction and 60 healthy control subjects to establish reliability and validity. The children were 24 to 61 months of age, with diverse ethnic and socioeconomic backgrounds. Strong internal consistency, significant correlation for test/retest reliability, moderate correlation for interrater reliability and support for external validity of the SSI was established. The SSI scores differentiated between the clinically referred subjects and healthy control subjects and between children with pervasive developmental disorder (PDD) of the autistic type and other non-PDD developmental disorders. The SSI is a relatively simple, efficient, reliable, and valid measure for the capacity for basic social interaction skills in children 24 months to 5 years of age. The SSI has a potential to be useful in primary health care settings to identify children at risk who may need tracking and/or further evaluation and treatment services. PMID- 9866089 TI - The pediatrician as therapist. II: Approaches to prevention of mental health problems and to behavior management. PMID- 9866090 TI - Extraordinary changes in behavior in an infant after a brief separation. AB - A previously healthy 8-month-old infant experienced a dramatic change in affect, motor patterns, and appetite after a brief (4-day) separation from her parents. She went from being a happy, interactive, and engaging child to being withdrawn, with limited vocalization, spontaneous motor activity, or interest in her environment. For the first 3 days after her parents returned home, she refused solid food and most of the formula or water she was offered. Over the following week, her appetite increased minimally. Her parents were concerned about her apathy, excessive quietness, and lack of interest in food, toys, or their own attempts to engage her in play. At the pediatrician's office, she was significantly less interactive and playful compared with previous visits. Her temperature, heart rate, and respiratory rate were normal. Although there were no objective signs of dehydration, her weight was 8 ounces less than the previous recorded weight 1 month ago. Results of the physical examination were normal, with the exception of neurological and behavioral findings. She had mild, generalized decreased tone and strength, with normal deep tendon reflexes, a sad facial appearance with sustained furrowing of the lines in her forehead, difficulty in eliciting a social smile, and limited spontaneous sounds. There was an absence of focal neurological findings, a flat open anterior fontanel, and a head circumference measurement consistent with previous recorded results. There were no cutaneous bruises, and findings of her funduscopic examination did not reveal retinal hemorrhage. Before the 4-day holiday, child care was provided almost exclusively by her mother at home. On only two previous occasions, at 4 and 6 months of age, this first child of 20-year-old high school graduate parents was cared for by a neighbor when the parents went out to a movie theater. She cried for about 10 minutes when the parents left on these occasions. Her mother stated that she felt anxious about leaving the child for several days, but she and her husband decided that they needed a break and planned the 4-day vacation. Child care was provided by the neighbor, who did not notice any unusual behavior or refusal to feed while in her care during the parents' absence. PMID- 9866091 TI - Learning about children from literature. AB - Which tells us more about children, behavioral science or the general literature? Actually, neither gave us much information until the last two centuries. Only since Rousseau's contributions in literature and Darwin's in science have accurate descriptions been offered. Now one can find presentations on the impact of physical characteristics and disabilities, family interactions, other social influences, and temperamental predispositions in both literature and science. The difference in treatment is that science describes in abstractions the general principles of human nature, while literature synthesizes vivid, coherent illustrations of whole, believable persons in all their complexities. Literature, moreover, has ventured into some areas that science has not. The complete pediatrician needs both sources of information for a well-rounded knowledge of children and their development. PMID- 9866092 TI - Identifying efficiently and economically operated hospitals: the prospects and pitfalls of applying frontier regression techniques. AB - This article examines whether policy makers can make immediate use of recently developed frontier regression techniques to identify efficient hospitals primarily for rate setting purposes. This new approach is applied to data on Florida hospitals for the period 1982-1993 to appraise how well it performs in gauging variations in hospital costs and efficiency. This appraisal suggests that frontier methods are quite promising, but that they may not yield sufficiently unambiguous results to serve the short-term needs of hospital regulators interested in scaling reimbursement rates to the efficiency with which hospitals deliver services. Other policy-relevant strengths and weaknesses of frontier techniques are also discussed. PMID- 9866093 TI - Access to medical care under strain: new pressures in Canada and Australia. AB - Health policy changes intended to achieve cost control in OECD countries run the risk of reintroducing financial barriers to health care. However, although the problems faced are similar, different countries are dealing with the situation in different ways. For example, Canada and Australia, which share many similarities, have taken quite different policy paths in the last decade: Canada has preserved universal access, whereas Australian policy is promoting a two-tier system through the provision of public subsidies for private insurance. The evidence is that country-specific factors such as institutional arrangements, attitudes, and values intersect with economic and financial factors to shape policy outcomes. Moreover, the Canadian and Australian experiences suggest that in relation to access issues, attitudes and values are the key policy determinants. PMID- 9866094 TI - Rural hospital mergers, antitrust policy, and the Ukiah case. AB - This article examines the antitrust issues in rural hospital mergers by focusing on an important antitrust case involving the merger of two small hospitals in Ukiah, California. A key issue in this matter was whether the geographic market served by the merger included a nearby larger city. The economic efficiency of small rural hospitals and the competitive implications of their mergers are examined in the context of the Ukiah case. Economies of scale are shown to be important for small rural hospitals and should mitigate any increase in price. The efficiencies defense is shown to be difficult to make even when economies of scale make the likelihood of efficiencies high. The financial difficulties of many rural hospitals, especially in areas where too many exist, mean that mergers such as this one in Ukiah often are an efficient way to keep these hospitals financially sound and accessible. The Ukiah case suggests the desirability of the merger guidelines that permit most mergers of small rural hospitals. PMID- 9866095 TI - A blueprint for linking academic oncology and the community. AB - The processes by which academic medicine will train the next generation of physicians and develop new knowledge have brought to the forefront the relationship between academic medical centers and community hospitals and practitioners. Over the past thirteen years, the Harvard Medical School Joint Center for Radiation Therapy (JCRT) has developed an integrated radiation oncology program designed to serve simultaneously the needs of the community, teaching hospitals, medical schools, and faculty. The structure and function of this program are described here, as are the challenges posed by the highly competitive health care marketplace. We believe that long-term vision should guide short-term goals. The success of academic-community collaborative programs depends not only on the good will and vision of the participants but also on the medical administration, academic leadership, policy makers, and politicians who define the principles and rules by which cooperation within the health care industry occurs. PMID- 9866096 TI - Experimental circular keratotomy for correction of corneal astigmatism. AB - PURPOSE: To investigate the effect of circular keratotomy depth and diameter on corneal astigmatism. METHODS: High astigmatism was induced in 25 human donor eyes by an anterior radial 7-0 silk suture across the corneoscleral limbus. With a 6.0, 6.5, 7.0, or 7.5 mm trephine, a 0.3 mm deep circular incision was made in 20 donor eyes. In 5 donor eyes 5 circular incisions were made with a diamond knife set at 0.1, 0.2, 0.3, 0.4, and 0.5 mm depths. RESULTS: The reduction of astigmatism between the 4 groups in which different trephination diameters were used showed no statistically significant difference (P > .1). The relationship between incision depth and reduction of astigmatism was statistically significant (P < .001). CONCLUSION: Artificially induced corneal astigmatism was reduced by a circular keratotomy in this donor eye model. Deeper incisions showed a greater effect. PMID- 9866097 TI - Scleral fixation for a phakic anterior chamber disc intraocular lens. AB - BACKGROUND: Angle-supported phakic anterior chamber intraocular lenses (IOLs) require a high degree of anterior vaulting, but this can cause an inadequate distance between the IOL and the cornea, especially during rubbing or squeezing of the eye. METHODS: A vaultless implant was designed by anchoring 3 supple hepatics through the anterior chamber into the sclera. Thus, a suspended rather than a wedged implant was achieved. RESULTS: The lens was implanted in 3 eyes of 2 patients in 1990. Good tolerance of the IOL was maintained over 7 years of follow-up. CONCLUSION: A suspended anterior chamber phakic IOL maintains an adequate position with lack of bowing and shows promising results. PMID- 9866098 TI - Corneal wound healing after laser in situ keratomileusis in rabbits. AB - BACKGROUND: The aim of this study was to characterize the cell biology of wound healing in rabbit corneas subjected to laser in situ keratomileusis (LASIK). METHODS: Rabbit corneas underwent LASIK with various multizone photoablations or only a lamellar keratotomy followed by repositioning of the flap. We looked for indications for an active wound healing process. Immunohistochemistry for the extradomain A cellular fibronectin (EDA-cFn) or tenascin (Tn) and routine histology were examined. RESULTS: Four days after LASIK or lamellar keratotomy followed by repositioning of the flap, epithelial plugs and prominent keratocytes as well as Tn and EDA-cFn immunoreactions-indicative of a wound-healing process appeared in the wound margins. Epithelial plugs were less conspicous, and prominent, presumably activated, keratocytes were no longer identified at the wound margin at 2.5 and 5 months after wounding. However, EDA-cFn and Tn immunoreactivities could still be observed. Only the stromal cells located in the periphery of the flap and in relatively close contact with the epithelium were surrounded by scar tissue expressing immunoreactivity for EDA-cFn or Tn. The central corneal stroma was devoid of scar tissue. CONCLUSION: Results indicate that the wound healing reaction after LASIK takes place only at the periphery of the microkeratome wound, leaving the central optical zone clear. PMID- 9866100 TI - Photorefractive keratectomy for -1.25 to -25.00 diopters of myopia. AB - BACKGROUND: We evaluated prospectively the efficacy, predictability, stability, and safety of photorefractive keratectomy (PRK) for myopia. METHODS: Three hundred sixty-nine eyes of 257 patients were treated with an Aesculap-Meditec MEL 60 excimer laser. Treated eyes were divided into 3 groups: low myopes (-1.25 to 6.00 D), 226 eyes; medium myopes (-6.10 to -10.00 D), 104 eyes; high myopes ( 10.10 to -25.00 D), 39 eyes. Follow-up at 12 months was available for 348 eyes (94%). RESULTS: One year after surgery the number of eyes within +/- 1.00 D of emmetropia was 182 (86.7%) for low myopes, 43 (40.5%) for medium myopes, and 12 (30.8%) for high myopes. Values for +/- 0.50 D were low: 142 (67.6%), medium: 29 (29.3%), and high: 9 (23.1%). Three eyes with low myopia (1.4%) and 5 eyes with medium myopia (5.1%) lost 2 or more lines of spectacle-corrected visual acuity. None of the high myopes lost 2 or more lines. Uncorrected visual acuity of 20/20 or better was achieved in 82 eyes (39%) with low myopia; 20/40 or better was achieved in 183 eyes (87.1%). Five eyes (5.1%) of medium myopes achieved 20/20 or better; 52 eyes (52.5%) with medium myopia achieved 20/40 or better. Zero eyes with high myopia achieved 20/20 or better; 11 eyes (28.8%) achieved 20/40 or better. CONCLUSION: Photorefractive keratectomy proved to be an effective method to correct myopia up to -6.00 D. For myopia greater than -6.00 D, good results were achieved in most eyes when myopia was less than -10.00 D, but efficacy and predictability decrease. To avoid systematic undercorrection, slight overcorrection must be attempted with the Aesculap-Meditec MEL 60 excimer laser for the treatment of myopia. PMID- 9866099 TI - Astigmatism outcomes following spherical photorefractive keratectomy for myopia. AB - BACKGROUND: Our aim was to examine the change in astigmatism after spherical photorefractive keratectomy (PRK). These effects are essential to optimizing photoastigmatic refractive keratectomy (PARK) to correct astigmatism to within fractions of a diopter. METHODS: We retrospectively reviewed 98 eyes of 178 patients with mild to moderate myopia and cylinder < or = 1.00 diopter (D) treated with spherical PRK (VISX 20/20 STAR excimer laser system); 31 eyes had epithelium removed mechanically with a blade and 67 eyes by a laser-scrape technique. RESULTS: Refractive astigmatism was reduced by greater than 0.25 D in 27 eyes (28%); refractive astigmatism was induced by greater than 0.25 D in 31 eyes (32%); the average vector-corrected difference between an eye's astigmatism before and after surgery was 0.01 +/- 0.52 D (P = .85). Eyes with high topographic astigmatism but low refractive cylinder before PRK showed an average of 0.07 +/- -0.60 D change in refractive cylinder after PRK. Refractive astigmatism of more than 0.25 D was induced in 16 eyes (44%) that received manual removal of epithelium versus 15 eyes (24%) that received laser removal of epithelium (odds ratio 2.51, P < .01). CONCLUSIONS: When using PRK for astigmatism correction, refractive cylinder before surgery rather than topographic astigmatism may be the most appropriate method for targeting the astigmatism correction, especially when the two values are discordant. An unpredictable mild increase or decrease in astigmatism may be expected. PMID- 9866101 TI - Radial keratotomy clear zone diameter errors. AB - PURPOSE: To examine the variability of achieved clear zone diameters following radial keratotomy and to determine if refractive outcomes are correlated to clear zone diameter errors. METHODS: Twenty-six patients with radial keratotomy (51 eyes) were studied retrospectively. The central clear zone diameter for each radial incision meridian was measured using a slit-lamp and a photograph. The achieved clear zone diameters were statistically compared to the intended clear zone sizes and to the refractive outcomes. RESULTS: Twenty-nine of the 51 eyes met various exclusion criteria. Ninety individual slit-lamp clear zone measurements were obtained for the remaining 22 eyes. Sixty-two of 90 (69%) achieved clear zone diameters were more than 0.25 mm different than the intended clear zone size. A maximum difference of +/- 0.80 mm from the intended clear zone marker size was observed. The average clear zone diameter error did not statistically differ between the three refractive outcome groups (undercorrection, emmetropia, and overcorrection). CONCLUSIONS: In selected cases of centripetal radial keratotomy, the achieved clear zone diameter can vary widely from the intended clear zone size despite extensive surgeon experience. PMID- 9866102 TI - Microbiological examination of bandage soft contact lenses used in laser refractive surgery. AB - BACKGROUND: Disposable soft contact lenses are known to be colonized by bacteria and play a key role in bacterial keratitis pathogenesis. Such lenses, commonly used after laser refractive surgery procedures in which postoperative corneal infiltrations are sometimes observed, are potentially a substrate for bacterial inoculation. This study evaluates the extent of such a contamination. METHODS: Sixty disposable lenses collected from 60 eyes of patients who underwent photorefractive keratectomy (PRK), photoastigmatic refractive keratectomy (PARK), or laser in situ keratomileusis (LASIK) for the treatment of myopia or hyperopia were collected under sterile conditions over 4 months and cultured in various media. Results were statistically analyzed and the correlation with clinical and epidemiological data was examined. RESULTS: Eleven (18.3%) of the examined lenses were contaminated with Staphylococcus epidermidis. No other bacteria or fungi were found. Contamination was significantly more common among female patients (P = .036). Correlation with the other clinical or operative parameters examined was statistically insignificant. CONCLUSIONS: Contamination was independent of the surgical procedure and females who were frequent users of eyelid cosmetics displayed higher contamination frequencies, suggesting that bacteria possibly originate from eyelid flora. The isolation of Staphylococcus epidermidis requires close postoperative surveillance, since it is a known cause of keratitis. Prophylactic postoperative treatment with tobramycin, gentamycin, or sulphonamides could be indicated. PMID- 9866104 TI - Evaluation of increased aqueous outflow after radial keratotomy in enucleated bovine eyes. AB - PURPOSE: We studied the efficacy of using radial keratotomy incisions as a surgical method of increasing outflow facility in enucleated bovine eyes. METHODS: Each freshly enucleated bovine eye (n = 23) was cannulated to a system measuring the rate of flow into the eyes. Preoperatively, corneal thickness measurements were determined for all treated eyes. Radial keratotomy was performed in 13 eyes and a control set of eyes (n = 10) had no surgical manipulation. The intraocular pressure was set at 16.9 mm Hg. A clear zone of 10 mm was used for each eye with the incision depth set at 100% of the pachymetric reading. One set of experimental eyes had incisions that remained in the clear cornea, and the other set had incisions that reached 2 mm beyond the limbus into sclera. Preoperative outflow measurements were performed on each eye. Postoperative measurements were made after 10 incisions and 20 incisions. All eyes were checked for perforation after surgery and discarded if a perforation was found. RESULTS: No eye included in the analysis was perforated. The preoperative (baseline) total flow rate was 27.90 +/- 3.72 mu 1/min at 16.9 mmHg. The flow rate increased by 23.5% (over baseline) with 20 radial keratotomy incisions. A paired t-test was used to statistically compare the outflow measurements before radial keratotomy and after the first and second set of radial incisions. The increases in flow were found to be statistically significant (P < .005) for both 10 and 20 radial incisions. CONCLUSION: Assuming the flow measurements (flow rate) in these experiments parallel outflow facility, radial keratotomy increases the outflow facility in enucleated bovine eyes. PMID- 9866103 TI - The shape of Bowman's layer in the human cornea. AB - PURPOSE: The aim of this investigation was to derive a mathematical model for Bowman's layer, the interface between the epithelium and stroma, in the human cornea. METHODS: The central epithelial thickness distribution within 14 normal human corneas was measured in vivo using high frequency ultrasonic digital signal processing with a measurement precision of 2 microns. The results per eye were averaged and incorporated into existing algorithms for the estimation of the shape of the anterior surface of Bowman's layer using terminology in accordance with Baker's equation. RESULTS: The average radius of Bowman's layer was 7.34 mm (SE +/- 0.17 mm). Descriptions of this boundary ranged from a steepening or prolate ellipse to a hyperbola. However, the typical Bowman's layer is hyperbolic with a shape factor, p = -0.22 (SE +/- 1.81). CONCLUSION: The results support previous cadaver studies where Bowman's layer was found to be steeper than the anterior corneal surface but disagree with the concept that the average Bowman's layer is akin to a prolate ellipse. The hyperbolic nature of the average Bowman's layer has the potential to influence the optical performance of the eye. PMID- 9866105 TI - Refractive results of radial keratotomy after 10 years. AB - BACKGROUND: We analyzed retrospectively the refractive and visual results of a cohort of patients who underwent radial keratotomy for myopia 10 years ago. METHODS: Radial keratotomies using centripetal incisions were performed by the same surgeon in 1986 and 1987 to correct myopia of -0.75 to -10.00 diopters (D). Refractive and visual results were evaluated at 1 month and on average 10 years after surgery. RESULTS: Thirty-nine patients (71 eyes) underwent radial keratotomy. Mean spherical equivalent refraction before surgery was -3.90 D and after surgery was -0.50 D at 1 month and +0.13 D at 10 years after surgery. At last examination, 41 eyes (43.7%) had a spherical equivalent refractive error within +/- 1.00 D of emmetropia (24 eyes [34%] +/- 0.50 D) and 32.4% of eyes were overcorrected by more than 1.00 D. Between 1 month and 10 years after surgery, a hyperopic shift greater than 1.00 D occurred in 34.1% of eves with a single procedure. Ten years after surgery, 59 eyes (83%) had uncorrected distance visual acuity of 20/40 or better (22 eyes [31%] with uncorrected visual acuity of 20/20 or better). CONCLUSION: This retrospective evaluation confirms that radial keratotomy is an effective procedure to correct myopic error, but the hyperopic shift produces an unstable, less predictable refraction in one-third of eyes at 10 years. PMID- 9866106 TI - Photorefractive keratectomy in eyes with congenital nystagmus. AB - BACKGROUND: We report 2 eyes of 2 patients with nystagmus and myopia who underwent photorefractive keratectomy (PRK) to correct myopia. METHODS: A 32-year old male patient with a baseline spectacle-corrected visual acuity of 20/40 had PRK with an Aesculap Meditec MEL 60 excimer laser with a suction ring system for correction of a spherical equivalent of -8.25 diopters (D). A 19-year old male patient with baseline spectacle-corrected visual acuity of 20/50 had PRK with an Autonomous Technologies T-PRK flying spot and tracking excimer laser to correct a spherical equivalent of -5.00 D. RESULTS: Twenty-seven months after PRK, the operated eye of the 32-year old patient had an uncorrected visual acuity of 20/32 and a spectacle-corrected visual acuity of 20/25. Six months postoperatively, the operated eye of the 19-year old patient had uncorrected and spectacle-corrected visual acuity of 20/50. CONCLUSION: Patients with nystagmus are eligible for PRK to correct their refractive error with these two laser systems. PMID- 9866107 TI - Laser in situ keratomileusis interface deposits. AB - BACKGROUND: A clinical investigation was undertaken to determine the origin of a brown interface deposit that developed in 15 eyes after laser in situ keratomileusis (LASIK). METHODS: The LASIK procedure was evaluated and agents that were thought to be related to these brown deposits in the interface were eliminated sequentially. Eyes were observed after LASIK for presence or absence of the deposit. RESULTS: Twenty-five consecutive LASIK eyes in which a methylcellulose sponge was eliminated as a protector of the hinge developed no similar interface deposits. CONCLUSION: The use of a dry methylcellulose sponge to protect the hinge may result in unexpected interface deposits after LASIK. PMID- 9866109 TI - Sterile interface inflammation after laser in situ keratomileusis: experience and opinions. PMID- 9866108 TI - Corneal hydration monitored by laser-induced breakdown spectroscopy. AB - BACKGROUND: Corneal hydration is an important factor in laser corneal ablation. In photorefractive keratectomy (PRK), corneal ablation rate and final ablation surface quality are strongly dependent on corneal hydration. We used a spectroscopic technique for monitoring corneal hydration during PRK. METHODS: Hydroxyethlymethacrylate (HEMA) was employed for corneal hydration modeling. Hydrated HEMA samples were irradiated with a pulsed Nd:YAG laser (1064 nm, 10 mJ/pulse, pulse duration 15 nsec). Successive emission spectra corresponding to different degrees of hydration were recorded on a gated optical multichannel analyzer. The weight of the sample and hence its water content was monitored during the entire procedure with a sensitive balance. One rabbit and one human cornea were used to demonstrate the spectral analogy between the model and corneal tissue. RESULTS: The most noticeable dependence on water content of the substrate was that of atomic emission lines of Ca at 393 nm and 396 nm. CONCLUSION: Plasma emission spectra exhibited significant dependence on sample hydration. This dependence can be used for estimation of water content of irradiated model material and real cornea. PMID- 9866110 TI - Herpes simplex keratitis after LASIK. PMID- 9866111 TI - Waterjet microkeratome--to set the record straight. PMID- 9866112 TI - Electrical properties of colonic smooth muscle in spontaneously non-insulin dependent diabetic rats. AB - Electrical properties of colonic smooth muscle were investigated in the Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a model animal for spontaneous non insulin-dependent diabetes mellitus (NIDDM), and the results were compared with those obtained from the Long-Evans Tokushima Otsuka (LETO) rat, a control of OLETF rat. At experiments (aged 60-80 weeks), blood glucose level was about 171 mg/dl in LETO rats and 370 mg/dl in OLETF rats. Feces in the colon were restricted to the proximal region in LETO rats and distributed widely in the whole colon in OLETF rats. In both LETO and OLETF rats, the circular smooth muscle strips of the isolated distal colon revealed two types of spontaneous electrical response, slow wave and transient hyperpolarization. The resting membrane potential was smaller in OLETF rats than in LETO rats by about 3 mV, but it was not positively related with the blood glucose level. The amplitude of hyperpolarization produced by noradrenaline (NA) was smaller in OLETF rats than in LETO rats. Transmural nerve stimulation evoked a non-adrenergic, non cholinergic (NANC) inhibitory junction potential (i.j.p.) in both LETO and OLETF rats; the amplitude of the i.j.p. was smaller in OLETF rats than in LETO rats, while the latency of the i.j.p. was longer in OLETF rats than in LETO rats. Thus, in the distal colon, NIDDM may cause a depolarization of the membrane, an attenuation of NANC inhibitory transmission and a reduction in reactivity of adrenoceptors to NA. These results suggest that the constipation appearing with diabetes mellitus involves dysfunction of both the enteric autonomic nerves and the smooth muscles in the colon. PMID- 9866113 TI - Pyloric opening and closure evaluated by means of strain gauge force transducers. AB - Recently, gastrointestinal motility research and various studies to measure motility have been conducted. However, it is not easy to measure the pyloric sphincter motility, especially its open and closed motility. There are few reports on the opening and closed motility of the pyloric ring. In this study, I examined the pyloric motility with strain gauge force transducers (SGTs). SGTs were implanted onto the antrum, pyloric ring and duodenum of a dog. In order to clarify the relationship between the opening and closed motility of the pylorus and SGTs recordings, the pyloric ring was observed with a gastrofiberscope while measurements were taken using the SGTs in anesthetized dogs. In addition, the pyloric ring was extended mechanically with a balloon and measurement were taken using the SGTs. In conscious dogs, the natural gastropyloroduodenal motility was recorded. Then, cisapride, erythromycin, and Leu13-motilin were administered in the interdigestive state and the motility was recorded. The pyloric opening and closed motility could be monitored using SGTs. The pyloric ring opened when the pyloric motility recorded using the SGTs showed a negative deflection. In addition, in phase III, when intense contraction of the antrum was observed, relaxation and opening of the pyloric ring could be observed. Erythromycin and Leu13-motilin induced phase III-like motility and relaxation of the pyloric ring. However, in cisapride-induced motility, relaxation of the pyloric ring was not observed. The pyloric ring opening and closed motility can be monitored using SGTs and this method is effective for the evaluation of the pyloric function. PMID- 9866114 TI - Ca(2+)-sensitivity of portal vein circular muscle from normotensive and hypertensive rats. AB - Contractile sensitivity to Ca2+ was studied in rat portal vein circular muscle isolated from normotensive Wistar Kyoto rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP). There was no difference in resting membrane potential between WKY and SHRSP. Elevation of extracellular K+ concentration induced similar increase in tension in preparations from WKY and SHRSP. High K(+) depolarized preparation developed tension increase when extracellular Ca2+ concentration was increased. The sensitivity to extracellular Ca2+ was similar in preparations from WKY and SHRSP. Permeabilized preparation contracted when free Ca2+ concentration was increased. Sensitivity of permeabilized preparation to Ca2+ was similar in preparations from WKY and SHRSP. These results indicate that regulation of contraction by Ca2+ was similar between WKY and SHRSP. Since there was no change in high K-induced contraction in intact preparations or Ca-induced contraction in permeabilized preparations, there seemed to be no difference in Ca2+ influx via voltage-dependent channels in portal vein circular muscle from WKY and SHRSP. PMID- 9866115 TI - The mechanics of vascular cell motility. AB - Alterations in vascular cell shape and motility occur during developmental processes and in response to injury. Similarly, during tumor vascularization and atherogenesis, endothelial and smooth muscle cells undergo motile and proliferative responses to extracellular cues. Recent inroads into our understanding of signal transduction have identified several candidate pathways by which the extracellular matrix- and growth factor-mediated stimulation of vascular cell motility may be mediated. The multiple and divergent extracellular stimuli that stimulate vascular motile responses may converge on the cytoskeleton via a family of ras-related GTPases. Biochemical analyses as well as examination of cytoskeletal dynamics in vivo indicate that actin polymerization at the forward aspects of spreading cytoplasm is capable of driving forward protrusion formation in the absence of a conventional actin motor. Actin polymerization at the plasma membrane of leading lamellae may be mediated both by de novo nucleation of actin filaments and the generation of free filament ends by uncapping the barbed ends of existing actin filaments. This review summarizes the most recent findings in extracellular-cytoskeletal-signal transduction, therein, providing a framework to explain the remarkable remodeling seen in the vasculature during developmental and disease-related processes. PMID- 9866116 TI - A beta-actin isotype is present in rat cardiac endothelial cells but not in cardiac myocytes. AB - OBJECTIVE: Active constriction in cardiac capillary endothelial cells (CCECs) is controversial. It is thought by many researchers that CCECs are not actively involved in constriction; others believe that these cells do contribute some of the force required for capillary constriction. Because actin is a major component of most contractile mechanisms responsible for changing cell shape, we compared two probes as potential monitors of actin distribution in CCECs in situ. METHODS: We used SDS-PAGE, Western blotting, electron microscopy, conventional epifluorescence and confocal microscopy to evaluate an antibody against non muscle beta-actin and fluorescently labeled phalloidin to detect actin in CCECs. RESULTS: Fluorescently labeled phalloidin detected actin in cardiac muscle and faintly in CCECs. The antibody probe detected non-muscle beta-actin CCECs. However, this actin isotype did not occur in cardiac myocytes. In endothelial cells non-muscle beta-actin was concentrated at the cell periphery, and arranged as bundles of fibers, not as typical stress fibers. CONCLUSIONS: Phalloidin is not suitable as a probe to monitor actin distribution in CCEC's in situ. The antibody is a potentially useful tool in monitoring the actin cytoskeleton and determining the capacity of CCECs for active vasoconstriction in vivo. Non-muscle beta-actin in CCECs is arranged in filament bundles which are not typical stress fibers. PMID- 9866117 TI - Effect of flow on hydraulic conductivity and reflection coefficient of rabbit mesentery. AB - OBJECTIVE: To measure the hydraulic conductivity and reflection coefficient for albumin, as defined by the Starling equation, in rabbit mesentery. METHODS: A section of rabbit mesentery was fixed between two chambers filled with lactated Ringer solution. Flow (Q) of Ringer solution was measured across the mesentery at driving pressures (delta P) between 1 and 6 cm H2O. Hydraulic conductivity was proportional to the slope of the Q-delta P curve. At constant delta P (approximately -2, -4, or -6 cm H2O), flow was measured at three different albumin concentration differences (0.5, 1, and 1.5 g/dl). Unstirred layer effects were minimized by magnetic stirrers. Reflection coefficient was the negative of the slope of the Q-delta pi curve divided by the slope of the Q-delta P curve, where delta pi was the albumin osmotic pressure difference. Hydraulic conductivity was measured before and after testicular hyaluronidase was added to the Ringer solution. RESULTS: There was no significant difference in conductivity per unit area (Lp) for the three different driving pressures. Hyaluronidase increased hydraulic conductivity significantly (p < 0.01) by 72 +/- 56%, indicating that hyaluronan and other glycosaminoglycans contributed to tissue fluid resistance. Reflection coefficient (sigma) increased from 0.02 to 0.14 as flow increased eightfold. CONCLUSIONS: The results suggest that the mesentery tissue provides little restriction to the passage of albumin. The increase in conductivity in the presence of hyaluronidase indicates that tissue hyaluronan and other glycosaminoglycans provide fluid resistance to bulk flow. PMID- 9866118 TI - Prediction of permeability-surface area product data by continuous-distribution pore models. AB - OBJECTIVE: To investigate the ability of continuous-distribution pore models to accurately predict permeability-surface area product (PS) experimental data in skeletal muscle. METHODS: Models having a water-only (WO) pathway and continuous distributions of microvascular transport-pathway sizes were fit to solute reflection-coefficient (sigma) experimental data (approximately 0.5-16 nm Stokes radii) obtained from skeletal muscle to determine optimal parameter values. Without further modification, these models were used to predict experimental PS values obtained from the literature for small solutes ranging in size from NaCl to inulin and for three proteins, alpha-lactalbumin, ovalbumin, and albumin (approximately 0.23-3.7 nm radii). The protein PSs were determined from fluorescent tracer-diffusion curves and a nonlinear model of tracer diffusion in the cat hindlimb preparation. The model's PS predictions were compared to those of a discrete-pore model previously developed and a fiber-matrix (FM) model. RESULTS: A log-normal (LN) continuous pore-size distribution plus WO-pathway model (three free parameters) fit the sigma data to within the 95% confidence intervals of each of eight solutes spanning a 32-fold size range and was nearly as close to the data as was the two discrete-pore plus WO-pathway model (four free parameters). Both models closely described the PS data for nine solutes spanning a 14-fold size range. The fit of a fiber-matrix plus WO-pathway model (three free parameters) to the sigma data was much poorer than for the other models. CONCLUSIONS: The LN and two-discrete-pore models accurately describe sigma and PS experimental data in cat and human skeletal muscle. Therefore, experimental data resulting from complex microvascular transport processes are well characterized by simple pore models. PMID- 9866119 TI - An absolute requirement for P-selectin in ischemia/reperfusion-induced leukocyte recruitment in cremaster muscle. AB - OBJECTIVE: To systematically examine a role for P-selectin in a model of striated muscle ischemia/reperfusion (I/R). METHODS: Ischemia was induced in the cremaster muscle of mice by occluding the main feeding arteriole for 30 minutes. Blood flow was then restored to allow for 60 minutes of reperfusion and leukocyte kinetics were assessed during the control period (before I/R) and at 5, 30, and 60 minutes of reperfusion. To study a role for P-selectin in this model, three different approaches were used: Wild-type animals received fucoidin (10 mg/kg, i.v.), an anti-P-selectin antibody (RB40.34; 20 mg/animal, i.v.) at 25 minutes of ischemia, or I/R was induced in P-selectin-deficient mice. RESULTS: Ischemia/reperfusion induced a rapid and significant increase in leukocyte rolling, adhesion, and emigration in wild-type mice. The I/R-induced increase in leukocyte rolling was transient, inasmuch as it was reduced by approximately 50% at 30 minutes of reperfusion, and returned to control levels by 60 minutes. Both fucoidin and an anti-P-selectin antibody completely prevented the I/R-induced increase in leukocyte rolling. The P-selectin-deficient animals exhibited absolutely no baseline leukocyte rolling, adhesion, or emigration. Furthermore, I/R did not induce any increase in leukocyte recruitment in the P-selectin-deficient animals over the first 60 minutes of reperfusion. CONCLUSION: The results from this study clearly illustrate that P-selectin is absolutely critical in both baseline and I/R-induced leukocyte infiltration in the murine-striated muscle. PMID- 9866120 TI - Potentiation of pulmonary arteriolar vasoconstriction to endothelin-1 by inhibition of nitric oxide synthesis in the intact lung. AB - OBJECTIVE: To observe pulmonary arteriolar effects of endothelin-1 (ET-1) in the intact lung and determine if constriction to ET-1 is potentiated by inhibition of nitric oxide (NO) synthesis. METHODS: In anesthetized male Sprague-Dawley rats with open chest, the lungs were ventilated with air through the lower trachea and in vivo responses of pulmonary arterioles were examined by video microscopy. Observations were made when the lungs were statically inflated with oxygen to a pressure of approximately 10 cm H2O for brief periods. A lens with a dipping cone was held at the pleural surface. ET-1 (10(-7)-10(-5)M; approximately 0.1 ml) was applied topically to the fluid layer under the dipping cone. RESULTS: ET-1 (10( 6)M) constricted parent arterioles 60 +/- 5 microns in diameter by 52 +/- 12% (range: 20-100%) and branches 45 +/- 3 microns in diameter by 36 +/- 4% (19-48%). Constriction persisted and there was a dramatic long-lasting decrease in flow. Alveolar walls quickly became pale, indicating reduced capillary perfusion. A lower concentration of ET-1 (10(-7)M) constricted (p > 0.05) parent arterioles 61 +/- 4 microns in diameter by 7 +/- 3% initially, and by 13 +/- 8% after 14 +/- 2 minutes, while smaller branches did not respond. In separate experiments, infusion of the NO synthase inhibitor L-NAME (1 mg/kg per minute), modestly (10 +/- 3%) decreased (p < 0.05) baseline parent arteriolar diameter from 72 +/- 7 microns to 64 +/- 5 microns. Branch diameter changed insignificantly from 42 +/- 7 microns to 38 +/- 7 microns. After L-NAME, ET-1 (10(-7)M) constricted (p < 0.05) parent arterioles by 17 +/- 4% initially and 40 +/- 14% after 14 +/- 2 minutes. Concurrently, branches constricted (p < 0.05) by 14 +/- 4% and 26 +/- 15%. CONCLUSIONS: Arterioles less than 80 microns in diameter were very responsive to ET-1, which could be a factor in altering pulmonary microvascular resistance. Inhibition of NO synthesis appears to potentiate constriction to ET 1. PMID- 9866121 TI - Effects of environmental stress on the architecture and permeability of the rat mesenteric microvasculature. AB - OBJECTIVE: This study was performed to determine the effects of environmental stress on the leakage to albumin and architecture of microvessels in the rat mesentery. METHODS: One group of rats (Group A, n = 6) were examined immediately upon arrival at the animal care facility. Groups B (n = 24) and C (n = 32) were housed in rooms with high and low personnel activity, respectively, for up to 7 weeks. Group D (n = 18) was housed in the high activity room for 2, 3, or 4 weeks followed by the low activity room. RESULTS: Rats in the low activity room for 3-4 weeks showed robust microvascular networks within 25% to 50% of the mesenteric windows (each window consisting of the tissue extending between two adjacent feeding arterioles in the mesentery), whereas rats in Group B only showed fragile vessels at the edges of the mesenteric windows within fat deposits. Groups A and C demonstrated little mesenteric fat and few fragile vessels, in contrast to group B. Group D showed increased mesenteric networks and decreased mesenteric fat as recovery progressed. The microvascular networks of 6 rats, randomly selected from Group C, showed few venular leaks following perfusion with fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA). Such leaks were abundant in the mesenteric microvasculature of 3 rats randomly selected from Group B. CONCLUSIONS: This study demonstrates that environmentally induced stress alters the architecture and leakage to albumin of the rat mesenteric microvasculature, and emphasizes the importance of closely monitoring the environment of experimental animals. PMID- 9866122 TI - [Perithelial cells around cerebral blood vessels]. PMID- 9866123 TI - [Premature aging: recent advances]. PMID- 9866124 TI - [Hydromyelia associated with spinal lipoma]. AB - We studied morphological changes of hydromyelia complicated with spinal lipoma of infants who were untethered. Since the MRI was introduced, early detection of spinal lipoma with lumbosacral skin abnormalities has become possible. We have experienced 44 surgical cases of spinal lipoma. Out of 36 such cases, 25 (69.4%) had hydromyelia, and hydromyelia of 9 patients was found to be of the terminal ventricle type. As many as 36% of spinal lipoma patients with hydromyelia concurrently had terminal-type hydromyelia, which was considered to fall under a specific category of congenital hydromyelia. At the stage of embryogenesis of the spinal cord, the caudal cell mass undergoes vacuolization, canalization and retrogressive differentiation, and during this process, the terminal ventricle with ependymallined cells becomes morbid. Though the morphological changes of hydromyelia after surgery as untethering were varied, hydromyelia of 9 patients became smaller after untethering, and hydromyelia of 8 expanded but attenuated afterwards, totaling 17 (68.0%). However, 5 had an expanding tendency and 3 did not show any morphological changes during the follow-up period (20.6-26.9 months) by MRI. PMID- 9866125 TI - [Difference between transcortical sensory aphasia following the left frontal lesion and transcortical sensory aphasia following the left posterior lesion]. AB - We assessed the difference between transcortical sensory aphasia (TCSA) following the left frontal lesions (F-TCSA) and TCSA following the left posterior lesions (P-TCSA). All the patients were right-handed and the 7 patients had the lesions in the only frontal lobe and the 10 patients had the lesions only in the left temporo-parieto-occipital regions. We administered pointing tasks, using 90 line drawings representing single nouns. We presented 6 line drawings a pointing board, and we used two kinds of pointing boards: one showed the line drawings each belonging to different categories (random categorized pointing task), the other showed the line drawings each belonging to only either two different categories (two categorized pointing task) and we presented 15 pointing boards each alternatively. The result was that regarding the patients of P-TCSA showed different number of correct answers between the random categorized pointing task and the two categorized pointing task with statistical significance. Regarding the patients of F-TCSA showed no difference between them. The results indicated that disturbance of P-TCSA on the pointing task was the disturbance of semantic process per se. And the disturbance of F-TCSA on the pointing task was that of not only semantic process but also the whole process including comprehending the presented words, searching the line drawings, comparing the line drawings with the presented word and final selection, which demanded persistent multiple memory process consistent with working memory. PMID- 9866126 TI - [Intraoperative CT imaging system using a mobile CT scanner gantry mounted on floor-embedded rails for neurosurgery]. AB - Many neurosurgeons prefer to use intraoperative computed tomographic (CT) scanning, when possible, to check whether there is residual lesion or unexpected bleeding. We report a practical intraoperative CT imaging system using a high speed CT scanner installed in the operating room along with a digitally controlled neurosurgical operating table. We designed a rail-track system to mobilize the CT gantry. The gantry is fixed onto a motorized carrier that can be moved smoothly on a rail-track embedded in the floor and with a maximum reach of 2.85 m from the room's wall to the operating table. The longitudinal motion of the operating table is easily adjusted by a foot switch from manual control to automatic control directly from the CT scanner's computer like an ordinary CT scanner bed in increments of 2, 5 or 10 mm during CT scanning. Either a carbon made radiolucent head frame or carbon-made head plate is used as a headrest. Using this CT scanner system, pre- and intraoperative CT scannings were performed on 46 patients with brain tumors, cervical lesions or other intracranial lesions. We could operate on the patient with enough working space between the mobile CT gantry and the operating table for microneurosurgery. We could obtain intraoperative CT imaging of a patient on the operating table while the surgical wound remained open, the surgical drapes kept in place, and the surgical position unchanged, saving time in intraoperative CT scanning and preparation for further surgery when needed. This intraoperative CT imaging system installed in the operating room should be useful for neurosurgery. PMID- 9866127 TI - [Cell to cell transfer of dense bodies induced by intracisternal infusion of leupeptin in rat cerebellum--quantitative and ultrastructural studies]. AB - Previous studies have demonstrated that rats treated with a proteinase inhibitor, leupeptin (Leu), show an accumulation of lipofuscin (Lf)-like dense bodies in the neurons. The present study examines the quantitative and ultrastructural changes in the Lf-like dense bodies of Purkinje (P) cells and Bergmann (B) glia over 20 days following a 4-day regimen of Leu administration by intracisternal infusion. Toluidine blue staining revealed considerable Lf-like granules in both types of cells 1 day after Leu infusion. However, these granules had almost disappeared by 20 days after infusion. Quantitative analysis revealed a decrease in the amount of accumulated Lf-like granules in P cells whereas the granules in B glia increased 10 days after Leu infusion. Electron microscopic examination revealed the presence of Lf-like dense bodies on the plasma membranes of both the P cells and B glia. The present results suggest that Leu-induced dense bodies in P cells are transferred to B glia, indicating a possible route for the removal of Lf. PMID- 9866128 TI - [Regional cerebral blood flow and cerebrospinal fluid amino acid analysis in elderly dementia]. AB - Regional cerebral blood flow and amino acid concentration in the cerebrospinal fluid were studied in 12 cases of vascular dementia, 12 cases of Alzheimer's disease, 12 cases of chronic alcoholism, and 12 age-matched healthy controls. In vascular dementia, blood flows were decreased in the cerebral cortex, frontal white matter, thalamus, caudate nucleus, and putamen and alpha-aminobutyric acid and glutamic acid concentrations were increased in the cerebrospinal fluid. In Alzheimer's disease, blood flows were decreased in the frontal cortex, parietal cortex, temporal cortex, and frontal white matter and alanine concentration was increased in the cerebrospinal fluid. In chronic alcoholism, blood flows were decreased in the cerebral cortex, thalamus, and putamen and urea, alanine, and glycine concentrations were increased in the cerebrospinal fluid. PMID- 9866130 TI - [A case of crushing head injury with bilateral abducens and facial nerve palsies]. AB - A 20-year-old male was admitted to our hospital suffering from a crushing head injury. At accident, his head had been compressed by the printing machine on both temporal regions. He remained at dull conscious. On admission one hour after the injury, he showed bilateral sixth-nerve and seventh nerve palsies and bleeding from the nose. CT scan showed marked pneumocephalus, traumatic subarachnoid hemorrhage, fluid collections in the bilateral sphenoid sinuses and right mastoid air cells. Bone CT disclosed bilateral temporal bone fractures. MRI did not show cerebral parenchymal damage. He recovered fully conscious at four hours after the injury, but cranial nerve palsies sustained over 30 days after the injury. Bilateral decompression of the facial canal were performed at day 31. At one year after the injury, bilateral abducens nerve palsies and facial nerve palsies recovered incompletely (grade III). The case report and the mechanism of such cranial nerve injuries by low-velocity crushing head injury is described. PMID- 9866129 TI - [Serial MRI, SPECT and 1H-MRS findings in a case of herpes simplex encephalitis]. AB - We report herein a case of herpes simplex encephalitis (HSE) in which magnetic resonance imaging (MRI), single photon emission computed tomography (SPECT) and proton magnetic resonance spectroscopy (1H-MRS) were performed sequentially. Both MRI and SPECT demonstrated a lesion on the left temporal lobe in its early stage. However, SPECT seemed more sensitive because it could detect subtle alterations of the blood flow. Hyperperfusion identified by SPECT had returned to normal levels while the abnormal MRI findings still remained. This may indicate that normalization of the blood perfusion precedes the tissue recovery. Contrast enhancement using Gd-DTPA was observed along the surface of the cerebral cortex on MRI, but that involved only part of the hyperperfusion areas depicted with SPECT. In the affected left temporal lobe, 1H-MRS exhibited reduction of N-acetyl asparate (NAA), and an elevated lactate level. The former represented the neuronal loss, and the latter indicated impairment of oxidative metabolism. Furthermore, alterations of NAA/creatine (Cr) and choline (Cho)/Cr ratios were also observed in the seemingly normal right temporal lobe. Those ratios returned to nearly normal levels in accordance with the patient's clinical recovery. Subsequently, our results indicate that 1H-MRS might be a very useful tool in qualifying the tissue damage and possibly estimating the prognosis. PMID- 9866131 TI - [Dural metastasis of prostatic carcinoma on the middle fossa]. PMID- 9866132 TI - [Bilateral middle cerebellar peduncle infarction]. PMID- 9866133 TI - [A 61-year-old man with progressive gait disturbance, freezing, and vertical gaze paresis who developed esophagus cancer]. AB - We report a 61-year-old Japanese man who died of complications of esophagus cancer surgery. He was well until his 55 years of the age, when he had an onset of speech disturbance and hand writing. He was seen by a neurologist who prescribed Menesit 600 mg/day. His symptoms improved with this medication. In 1993, three years after the onset, he started to show gait disturbance and easy to fall. In 1995, he noted difficulty in eye opening. He visited our clinic on October 26, 1996. On examination, he showed vertical gaze paresis, masked face, nuchal rigidity, small step gait, freezing phenomena, and festination. His mental status was normal. He was treated with 800 mg/day of Menesit, 800 mg/day of L dops, and 10 mg/day of bromocriptine with little improvement in his symptoms. Cranial CT scan revealed some dilatation of the third ventricle. Subsequent clinical course was one of the slow progression of his parkinsonism. In September of 1997, he noted difficulty in swallowing. He was admitted to the gastrointestinal service of our hospital on October 14, 1997. On admission, neurologic status was essentially similar to the previous one, but he showed more advanced state of his parkinsonism. Upper gastrointestinal series revealed a mass lesion of about 11.5 cm in length protruding into the lower esophagus lumen. Subtotal esophagus resection including the mass was performed on December 2, 1997. The stomach was elevated for anastomosis with the upper esophagus. No metastases were found in the mediastinum except for two lymph nodes in the para esophageal region. The subsequent course was complicated by marked elevation of GOT, GPT, LDH, total bilirubin as well as direct bilirubin, alkaliphosphatase, and amylase starting in the evening of the surgery. On December 7, leukocytosis and pneumonic shadow were seen involving his right lung. On December 10, he developed cardiopulmonary arrest. He was once resuscitated; however, he developed cardiac arrest again seven hours later and pronounced dead. He was discussed in a neurologic CPC. The chief discussant arrived at the conclusion that the patient had PSP and the cause of the death was ascribed to circulatory disturbance to the liver. The discussant also thought that the terminal course was complicated by cholangitis or cholecystitis, sepsis, and pulmonary embolism. Surgical specimen of the esophagus tumor revealed carcinosarcoma. Postmortem examination revealed yellowish discoloration of the peritoneum and mesenterium, and accumulation of clouded ascites indicating the presence of peritonitis. Inflammatory change extended to the mediastinum. On microscopic examination, various kinds of bacilli and candida spores were seen. The liver was enlarged and a perforation was noted in the gallbladder causing biliary necrosis in the adjacent liver. An extensive infarct was seen in the left lobe of the liver; this was found to be due to obstruction of the hepatic artery at the site of the duodenohepatic mesenterium and obstruction of intrahepatic portal vein secondary to retrograde intrahepatic cholangitis in the left lobe. A piece of surgical threads was seen adjacent to the hepatic artery; foreign body granulomatous reaction was seen surrounding the surgical thread. The rupture of the gallbladder appeared to be due to the obstruction of the left branch of the hepatic artery. Neuropathologic examination revealed extensive degeneration of the pallidum, the substantia nigra, and the subthalamic nucleus and presence of neurofibrillary tangles in the remaining neurons. The neuropathologic findings were consistent with progressive supranuclear palsy, although the pathologic changes in the midbrain tegmentum was only mild gliosis. PMID- 9866134 TI - A pediatrician's view. Why do we have paranasal sinuses? PMID- 9866135 TI - Sinusitis overview. PMID- 9866136 TI - The common cold. PMID- 9866137 TI - Allergic rhinitis. PMID- 9866139 TI - The surgical management of chronic sinusitis in children. PMID- 9866138 TI - Sinusitis. PMID- 9866140 TI - In search of perfection. PMID- 9866141 TI - American Heart Association guidelines. PMID- 9866142 TI - Pediatric bone marrow transplantation: oral complications and recommendations for care. AB - Bone marrow transplantation (BMT) has become an increasingly common treatment option for those patients who have a disease that affects the bone marrow (BM) directly or indirectly. Because of the level of immunosuppression achieved in BMT, any problems the pediatric patient presents in the oral cavity can become life-threatening and increase the length of hospital stay, the patient's discomfort, and the treatment costs. The oral cavity is a reservoir for microorganisms thus by reducing their number through optimal care, immunosuppressed patients may decrease their chance of a life-threatening systemic infection from an oral source. Many BMT teams believe that toothbrushing increases the risk of bacteremia and bleeding; however, problems are more likely to arise when patients are not compliant with good oral hygiene habits. It is vital to educate the caretaker and the child about the importance of oral care in order to minimize discomfort and maximize the chances for a successful transplant. This paper discusses the important and unique role that pediatric dentistry has in the multiprofessional BMT team to help bring about a successful outcome through the prevention and treatment of the acute oral complications often seen in these patients. PMID- 9866143 TI - Estimation of the caries-related risk associated with infant formulas. AB - PURPOSE: Baby bottle tooth decay (BBTD) affects 6% of children under three years of age and is associated with inappropriate bottle use. The objective of this study was to estimate the caries-related risk associated with 26 infant formulas and whole milk. METHODS: First, the plaque pH of adult volunteers was monitored before and after an oral rinse with infant formula to determine the minimum pH obtained in response to each formula. Second, Streptococcus sobrinus 6715 was cultured in each infant formula, and the increase in the number of colony forming units was measured. Third, each infant formula was incubated with powdered enamel and the solubility of enamel mineral was calculated in the absence of bacteria. Fourth, each formula was mixed with standardized concentrations of acid to determine the buffering capabilities. Finally, enamel windows were created on extracted permanent molars and exfoliated primary incisor crowns that were then colonized with mutans streptococci and incubated with infant formula. Caries was assessed visually and radiographically for 18 weeks. The length of time required for the development of enamel caries, dentinal caries and pulpal involvement was recorded. RESULTS: One-way or two-way ANOVA of these five assays demonstrated that 1. Plaque pH varied in response to oral rinsing with infant formula and most formulas did have the ability to reduce the pH significantly below the pH obtained after rinsing with water 2. Some infant formulas supported significant bacterial growth 3. Enamel mineral was dissolved by incubation with certain infant formula 4. The buffer capacity varied among the infant formulas tested 5. The length of time required for caries to reach dentin or pulp differed for the formulas, with some formulas causing dentinal caries by 3.4 weeks and pulpal involvement by 7.2 weeks. PMID- 9866144 TI - A retrospective study of oral and maxillofacial biopsy lesions in a pediatric population from southern Taiwan. AB - This is a retrospective study of biopsied oral lesions (N = 534) in a pediatric population (0-15 years old) in southern Taiwan. In this study, we included 6% of the 9672 biopsies performed from 1985 through 1996. The lesions were divided into three groups according to patients' ages, 0-5 years old; 6-10 years old; and 11 15 years old The majority of the samples were from the oldest age group (273/518). With the exclusion of normal tissues (N = 26), 518 samples were classified into four categories: inflammatory lesions, cystic lesions, tumor or tumor-like lesions, and other lesions. The largest number of lesions occurred in the inflammatory lesion group (46%). The 12 most frequently occurring lesions contributed about 78% of all the biopsies in the three age groups (0-15 years old). Frequencies of the lesions of mucous extravasation phenomenon, dentigerous cyst, fibrous dysplasia, and odontonia in our three pediatric age groups showed a significant proportion in the biopsies of the same lesions in the group of patients of all ages. These information may be valuable for both epidemiology and teaching. PMID- 9866145 TI - In vitro enamel effects of a resin-modified glass ionomer: fluoride uptake and resistance to demineralization. AB - PURPOSE: The purpose of this study was to determine the in vitro effects of a resin-modified glass-ionomer restorative material on human enamel. METHODS: Enamel specimens were restored with Photac-Fil, a resin-modified glass ionomer; Ketac-Fil, an autopolymerizing glass ionomer and silver amalgam. The samples were pH cycled and then subjected to an artificial caries challenge. Specimens were evaluated by fluoride microdrill biopsy and quantitative microradiography at 1, 2, and 3 mm from restorations. Data on fluoride content, lesion depth, and mineral content were analyzed by one-way ANOVA, Student's t test, Neuman-Keuls procedure (multiple [pairwise] comparison), and Pearson's product-moment correlation tests. RESULTS: There were significant differences in fluoride uptake, lesion depth, and mineral density between groups. Significant correlational relationships were also determined. CONCLUSION: This study demonstrates that Photac-Fil and Ketac-Fil prevent in-vitro demineralization of enamel and suggests different mechanisms of action for each material. PMID- 9866146 TI - Evidence-based dental care--a concept review. AB - Evidence-based (science-based) care is the prevailing paradigm in the health sciences. This paper briefly reviews the basic concepts of evidence-based care. Assessment of scientific evidence including information retrieval, evaluation of the quality of evidence, and finally the synthesis of collected evidence is described. Practitioner approach to evidence-based care through the twin modalities of literature reviews and clinical practice guidelines is outlined. It is concluded that the promotion of outcomes research and the adoption of evidence based practice is likely to improve the delivery of a high standard of quality dental care. PMID- 9866147 TI - The reinforced laryngeal mask airway (LMA) as an alternative airway device to manage the difficult airway. PMID- 9866148 TI - Apexification of a primary central incisor: 6-year follow-up. PMID- 9866149 TI - Peripheral ossifying fibroma associated with a neonatal tooth: case report. PMID- 9866150 TI - [Prophylaxis of neurological manifestations of lumbar osteochondrosis: intermediate results, unsolved problems and some methodologic aspects]. AB - Issues of prophylaxis and rehabilitation are considered as regards neurologic manifestations of spinal osteochondrosis (NMSO). NMSO primary prophylaxis requires investigation and quantitative evaluation of the disease risk factors, epidemiologic determination of both incidence and prevalence in different groups of population in various periods of time. The standard risk group of NMSO in Byelorussian consists of population of 40-49-year-olds. Some professions may be risk factors for both onset and relapse of NMSO. Secondary prophylaxis of NMSO concerns elevated relapse risk, screening for individuals with either subclinic or early clinic manifestations of the disease. The computer screening-system has been created to identify a group at high risk of NMSO onset or relapse, to keep records, to print documents. It also gives chance to carry out individual prophylaxis of the high risk in the course of mass examination of employees. PMID- 9866152 TI - [Types of impulse control disorders in children and adolescents]. AB - The paper presents the results of examination of 61 patients aged 7-15 years with impulse-control disorders (drive) within psychopathy-like syndrome in slow progredient schizophrenia or in residual-organic disorders of CNS. 4 types of impulse-control disorders were identified: 1) drives concerning mainly instincts' disorders (enhanced physiologic drives, perversive physiologic drives); 2) drives due to disorders of will (the impulsive drives, the precursors of the impulsive drives); 3) the obsessive drives; 4) "social" forms of altered drives. In the group of schizophrenic patients prevailed disturbances of drives concerning the instinctive sphere (97.9%) and seldom "social" forms (2.1%) were ovserwed. In the patients with residual-organic CNS damage the "social" forms were quite frequent (50%) as well as disorders of will (45.8%), but disorders of instincts (4.2%) occurred rarely. Children and adolescent had often incomplete drives in the form of precursors of the impulsive drives. PMID- 9866151 TI - [Clinical-neuropsychological aspects of the adaptation of patients after the excision of craniopharyngioma in childhood]. AB - 40 patients aged 9-36 years were followed-up for at least 10 years after craniopharyngiomectomy. A combined neuropsychologic study according to A.R. Luria was carried out both for determination of the degree of psychic defects and for comparison with possibilities of adaptation. Symptoms of a damage of medio-basal regions of the frontal lobes and dyencephalic region were observed in all the patients. The frontal symptoms were decisive for social adaptation after the operation. The adaptation was worse in patients with more pronounced manifestations of frontal symptoms in involvement of medio-basal regions of the left frontal lobe accompanied by poor motivation and inertness. Patients with infact motivation and activity in the presence of dominant dysfunction of the medio-basal regions of the right frontal lobe appeared to be more adapted, even when there were pronounced disorders of vision and memory. The defects were compensated worse in patients operated early in childhood, with features of side cerebral dysfunction and in left-handers. PMID- 9866153 TI - [Paranoid nosogenic reactions in patients with somatoform disorders]. AB - The authors examined 16 patients with ischemic heart disease who had psychogenic disorders with ideas of rationalization of therapy. Paranoial personality disorders (paranoial psychopathy) was diagnosed in 11 patients, pseudopsychopathy in residual schizophrenia in 5 patients. The patients had either a fanatic idea of creating new methods of self-therapy and an autoaggressive behavior (performing some dangerous manipulations, usage of non-medicamentous substances, etc.). The premorbid state of such patients was characterized by inclination to paranoial reactions and a special predisposition to them combined with an egodystonic attitude to corporal manifestations. Paranoial reactions of such type were reversible and disappeared in the course of somatic state improvement. PMID- 9866154 TI - [The use micro-polarization in spinal cord lesions]. AB - Transdermal micropolarization of the spinal cord was made in patients with consequences of the spinal cord injury or tuberculous spondylitis. Changes in clinical and electrophysiologic status were evaluated. It was found that local direct current through dermal electrodes promotes an improvement of both motor and autonomic functions in such patients. This corresponded to a positive dynamics both of the spinal cord state and cardiac activity. Possible mechanisms of influence of the direct current on the spinal cord as well as perspectives of application of micropolarization in spinal cord's damage are outlined. PMID- 9866156 TI - [The role of nuclear magnetic resonance monitoring in early clinical-instrumental diagnosis of multiple sclerosis]. AB - Nucleo-magnetic resonance tomography (NMRT) facilitated considerably early diagnosis of multiple sclerosis. Yet, there appeared data indicating the existence of NMRT negative forms. These were found in 15 patients of 82 ones examined using NMRT only which failed to defect foci of demyelinization in spite of a clear-cut clinical picture. NMRT monitoring during 3 years (from 2 to 5 NMRT investigations) permitted to visualize the foci of demyelinization located primarily in the periventricular cerebral region. The conclusion was made that a presence of clear clinical symptoms as well as a remitting character of the pathologic process confirmed by visualization of multiple foci of CNS demyelinization by means of NMRT monitoring, gave full chance for a reliable diagnosis of multiple sclerosis. A lack of such confirmation may be explained by the fact, that clinical debut of the disease reflected only the phase of multiple sclerosis and forestalled forming of a focal demyelinizating process, pathognomonic for this disease. PMID- 9866155 TI - [Individual computer-assisted pharmacotherapy in Parkinson disease]. AB - The authors examined whether it is valid to use electromyographic signs for control of both the efficiency of pharmacotherapy and an individual choice of drugs. A new computer method of processing of muscular electrical potentials' registered-by supradermal electrodes is proposed. The method permitted quantitation of electromyographic signs of the main symptoms of Parkinson's disease (tremor, rigidity, akinesia). Based on the signs obtained, a system of automatic diagnosis of the main symptoms of Parkinson's disease by a separate muscle was designed. This method was used in examination of 72 patients with different clinical forms of Parkinson's disease in the course of selectivity pharmatherapy for them. The method can be used for more precise diagnosis of the disease of clinical forms, for an individual choice of drugs during long-term antiparkinsonic mono- and combined therapy and for evaluation of efficiency of the treatment performed. PMID- 9866157 TI - [Association of allele polymorphism of dopamine d2 receptors with schizophrenic and affective disorders]. AB - DRD2 Taq polymorphism has been studied in the samples from 214 patients (119 males, 95 females, mean age 37.8 +/- 13.6) and of 96 controls (50 males, 46 females, mean age 40.7 +/- 20.0). The latter group comprised 75 unrelated controls and 21 healthy first degree relatives of schizophrenic patients. All the patients were diagnosed according to ICD-10 and have been divided into 4 groups: paranoid schizophrenics (n = 102), schizotypic patients (n = 25), patients with schizoaffective psychoses (n = 40) and affective disorders (n = 47). Taq1A DRD2 polymorphism was represented by 3 genotypes: A1A1, A1A2, A2A2 (allele A2 was a result of nucleotide substitution). The frequencies of genotypes in affected group didn't significantly differ from a control one. However, a frequency of A2A2 genotype (0.45) in a group of paranoid schizophrenics was significantly higher, than in the patients with schizoaffective psychoses (0.22) or in a control group (0.26), but was similar to that of the patients with schizotypic or affective disorders (0.4). A2A2 DRD2 genotype seems to be a potential genetic factor of susceptibility to schizophrenia. PMID- 9866159 TI - [Mediated short-term influence of alternating magnetic field on the epileptic brain]. AB - The changes of bioelectric activity of the brain hemispheres were studied under the influence of an alternating magnetic field (AMF), directed on the hand of 20 epileptic patients and 18 healthy normals. Under AMF influence there was intensification of epileptic activity--elevation of the focal bioelectric characteristics and of the processes of generalization. It permitted to apply AMF for diagnosis of epileptic focus. Under AMF influence more stable changes in posterio-parietal areas of the right hemisphere occur. The authors came to the conclusion that AMF effects were mainly realized with participation of right hemisphere. PMID- 9866158 TI - [Clinical and neurophysiological aspects of somatosensory afferentation system in patients with hemispheric ischemic stroke]. AB - Method of somatosensory evoked potentials (SSEP) was applied for examination of 36 patients in early restorative period of ischemic stroke located in internal capsule, in cortical-subcortical area of parieto-temporal cerebral regions or in cortical-subcortical areas of frontal areas. 15 healthy individuals formed a control group. Electrophysiologic SSEP pattern was not homogeneous and depended on the location of a damage focus in cerebral hemispheres. Peculiarities of generatory mechanisms of SSEP components' forming in ischemic stroke, are presented. PMID- 9866160 TI - [The prophylaxis of cerebral vascular diseases]. PMID- 9866161 TI - [Psychophysiological health aspects of population living in radiation contaminated territories]. AB - A combined psychophysiologic study of 100 individuals living in territories contaminated by radionuclides and 60 people from ecologically clean region. There were no significant differences in the incidence of neurotic disorders between people from these regions (61.2 and 65.3%, respectively). The highest sensitivity to small radiation doses was observed at rather young age (20-40 years). It was found that a considerable role in the development of borderline psychic disorders belongs to both somatic disturbances due to low-dose radiation and psychogenic effects. A combined action of psychogenic and somatogenic factors aggravated symptoms of neurotic disorders and promoted widening of a range of psychic disturbances. Some part of the population from the contaminated territories tended to the development of cerebro-vascular psychoorganic pathology resultant from a breakdown of the reserves of the adaptation. Now the stress factor of radionuclides exposure is not so important as social and economic problems. PMID- 9866163 TI - [Modern approaches to the therapy of Alzheimer's disease]. PMID- 9866162 TI - [The formation and development of clinical neurology in Imperial Medical-Surgical (Military-medical) academy: on the 200th anniversary of the Military-Medical Academy and on the 100th anniversary of the Clinic of nervous diseases]. PMID- 9866164 TI - The adaptation of existing personal inhalable aerosol samplers for bioaerosol sampling. AB - Health-related monitoring of bioaerosol exposures in the workplace should ideally be carried out using size-selective personal samplers that separate the aerosol into biologically relevant size fractions and allow both quantification and identification of the microorganisms present in each fraction. As a first stage in the development of personal bioaerosol samplers a number of collection substrates were assessed for their ability to maintain the viability of the collected microorganisms, so that subsequent culturing and species identification may be carried out. The substrates were tested with bioaerosols of varying robustness, consisting of Saccharomyces cerevisiae cells, Escherichia coli cells, and Penicillium expansum spores, aerosolized under controlled environmental conditions. The survival of microorganisms on each test substrate, assessed on the basis of the culturable fractions of cells recovered, was compared with that of microorganisms collected in a reference glass cyclone sampler. These bioefficiency tests identified the substrate combinations with the potential to fulfill personal sampler design criteria. The substrates were then subjected to further development to evaluate and optimize their particle size selection characteristics. The outcome of this work is two prototype personal bioaerosol samplers in which size-selective substrates are adapted for use in existing designs of personal inhalable sampler. This offers an effective and low-cost solution to personal monitoring of bioaerosol exposures in the workplace. PMID- 9866165 TI - Comparison of six respirator fit-test methods with an actual measurement of exposure in a simulated health care environment: Part I--Protocol development. AB - Quantitative fit tests (QNFT) have been assumed to be predictive of the protection respirators would provide to a wearer in the workplace. Workplace studies have consistently found no correlation between quantitative fit factors and workplace protection factors. This article is the first in a series of three describing a study designed to compare the fit factors from six QNFT methods against the actual dose of 1,1,2 trichloro-1,2,2 trifluoroethane (Freon-113) received under the same laboratory conditions. Five preliminary studies conducted to develop the protocol to assess the respirator wearer's dose through end exhaled air analysis are described in this article: (1) chamber characterization, (2) end-exhaled air sampling, (3) skin absorption testing, (4) pharmacokinetic modeling, and (5) subject characterization. It was established that the concentration of corn oil aerosol and Freon-113 could be generated simultaneously in the chamber. It was ascertained that the optimum time to sample the exhaled breath was 30 minutes after the subject exited the chamber. It was also found that in a chamber concentration of 500 ppm, without any respiratory exposure, Freon-113 was still present in the end-exhaled air. This was attributed to skin absorption. The end-exhaled air of subjects exposed to 0.5, 3, 5, 25, 50, and 100 ppm (30 minute time-weighted average) of Freon-113 was evaluated at 30 minutes postexposure. This characterization was then used to predict the actual dose of Freon-113 received during the method comparison and validation testing to be described in subsequent articles. PMID- 9866166 TI - Comparison of six respirator fit-test methods with an actual measurement of exposure in a simulated health care environment: Part II--Method comparison testing. AB - This article, the second in a series of three, describes the method comparison testing portion of a study conducted to compare the fit factors from six quantitative fit-tests (QNFT) with a measure of a respirator wearer's actual exposure assessed by end-exhaled air analysis for 1,1,2-trichloro-1,2,2 trifluoroethane (Freon-113) under the same conditions. The six QNFT methods were (1) continuous low flow, flush probe; (2) continuous high flow, deep probe (CHD); (3) exhalation valve discharge (EVD); (4) controlled negative pressure; (5) 10 minute Ambient Aerosol 1 (AA1); and (6) 30-minute Ambient Aerosol 2. The first three methods utilized corn oil and a forward light scattering photometer. The last two methods used the TSI Portacount. Respirators used in the study were both disposable and elastomeric organic vapor/high efficiency half-masks. The characterization equations from the preliminary research (described previously) were used to determine the actual exposure to Freon-113 during the method comparison testing. The fit factors resulting from the QNFT methods were then individually correlated with the Freon-113 exposures using the coefficient of determination, R2. The lowest R2 value, 0.20, was found with the EVD method. The highest R2 values, 0.81 and 0.78, were associated, respectively, with the CHD and AA1 methods. This study suggests that some QNFT methods may be used to estimate actual respirator performance under laboratory conditions. PMID- 9866167 TI - Methods for assessing the physical demands of manual lifting: a review and case study from warehousing. AB - Assessment of the physical demands of potentially hazardous manual material handling (MMH) activities is fundamental to the prevention of disabilities from occupationally related low back pain, a problem costing the nation billions of dollars annually. Although there is a variety of ergonomic assessment methods available for assessing MMH activities, there is a lack of practical information to assist users in choosing the most appropriate assessment methods of a particular job. This article reviews currently available assessment methods and presents case study results of a physically demanding repetitive manual lifting job in two grocery warehouses. The case study will provide a framework for a comparison of the methods and a discussion of relevant application issues designed to assist users in selecting appropriate methods for assessing MMH jobs. Based on the results of the study, it is concluded that all of the ergonomic methods were in agreement that the job of grocery selector has a high level of risk for low back pain. Differences between the methods were noted, however, that should be considered when choosing a specific method for a specific application. PMID- 9866168 TI - Evaluating the relative status of health and safety programs for minority academic and research institutions. AB - The health and safety programs that support academic and research institutions face many challenges because of the wide variety of potential hazards present and the possibility of simultaneous exposures to several agents. This study investigated whether differences in health and safety programs exist between minority and nonminority state-funded academic and research institutions. A cross sectional mail survey was conducted that included 24 minority and 51 nonminority schools. Questionnaires solicited information on descriptive institutional parameters, health and safety staffing, the hazards present, programs in place to address hazards, and medical surveillance services. The hazard types identified as present on both campus types were found to be very similar. The mean number of health and safety staff serving minority institutions was lower (1.14 versus 3.12), with the difference reliably predicted by the number of institutional employees, not minority status or regulatory environment. Other targeted parameters were found to be similar, with a consistent lack of awareness of specific medical evaluations noted for both groups. Since on average a single person is charged with controlling a diverse set of potential hazards on the minority campuses studied, the need for a comprehensive awareness of a variety of health and safety issues is discussed. PMID- 9866170 TI - Synthesis of biosurfactants in extreme conditions. AB - The interest in industrial biotechnology and its importance opens up challenging possibilities of research in this area. Surfactants have long been among the most versatile of process chemicals. Their market is extremely competitive and manufacturers will have to expand their arsenal to develop products for the year 2000 and beyond. Biosurfactants are one of the most promising compounds in this regard. A review of the literature reveals that studies on oil-degrading and biosurfactant-producing microorganisms deal almost exclusively with their synthesis in moderate environments. Biosurfactants and the microbes that produce them have numerous industrial, medical and environmental applications, which frequently involve exposure to extremes of temperatures, pressure, ionic strength, pH and organic solvents. Hence, there is a continuing need to isolate microbes that are able to function under extreme conditions. There is an urgent need to explore these extremophiles for their ability to produce biosurfactants that can function suitably under the conditions prevailing when they are applied. PMID- 9866171 TI - Production and structure elucidation of di- and oligosaccharide lipids (biosurfactants) from Tsukamurella sp. nov. AB - The bacterium Tsukamurella sp. nov., isolated from soil, was found to produce novel glycolipids when grown on sunflower oil as the sole carbon source. The glycolipids were isolated by chromatography on silica columns and their structures elucidated using a combination of multidimensional NMR and MS techniques. The three main components are 2,3-di-O-acyl-alpha-D-glucopyranosyl-(1 1)-alpha-D-glucopyranose, 2,3-di-O-acyl-beta-D-glucopyranosyl-(1-2)-4,6-di-O-acyl alpha-D- glucopyranosyl-(1-1)-alpha-D-glucopyranose and 2,3-di-O-acyl-beta-D glucopyranosyl-(1-2)-beta-D-galactopyranosyl- (1-6)-4,6-di-O-acyl-alpha-D glucopyranosyl-(1-1)-alpha-D- glucopyranosyl which are linked to fatty acids varying in chain length from C4 to C18. The glycolipids are mainly extracellular but are also found attached to the cell walls. During the cultivation the composition of the glycolipids changed from disaccharide- to tri- and tetrasaccharide lipids. The glycolipids show good surface-active behaviour and have antimicrobial properties. PMID- 9866169 TI - Oxidized resin acids in aerosol derived from rosin core solder. AB - Exposure to rosin during a variety of uses has been associated with dermal and pulmonary sensitization. Oxidized resin acids are present in many rosin products, and have been regarded as the main sensitizing rosin compounds in cases of dermal sensitization. This research describes oxidized resin acids identified in aerosol produced during soldering with rosin core solder. Oxidized resin acids found were 7-oxodehydroabietic acid, 15-hydroxydehydroabietic acid, and 7 hydroxydehydroabietic acid. The presence of oxidized compounds known to be dermal sensitizers in aerosol from rosin flux soldering supports the hypothesis that resin acid compounds are pulmonary sensitizers as previously proposed. Changes in the composition of resin acid aerosol derived from heated rosin core solder (compared with the parent material) are described. PMID- 9866172 TI - Evaluation of cell recycling in continuous fermentation of enzymatic hydrolysates of spruce with Saccharomyces cerevisiae and on-line monitoring of glucose and ethanol. AB - The maximum growth rate of Saccharomyces cerevisiae ATCC 96581, adapted to fermentation of spent sulphite liquor (SSL), was 7 times higher in SSL of hardwood than the maximum growth rate of bakers' yeast. ATCC 96581 was studied in the continuous fermentation of spruce hydrolysate without and with cell recycling. Ethanol productivity by ATCC 96581 in continuous fermentation of an enzymatic hydrolysate of spruce was increased 4.6 times by employing cell recycling. On-line analysis of CO2, glucose and ethanol (using a microdialysis probe) was used to investigate the effect of fermentation pH on cell growth and ethanol production, and to set the dilution rate. Cell growth in the spruce hydrolysates was strongly influenced by fermentation pH. The fermentation was operated in continuous mode for 210 h and a theoretical ethanol yield on fermentable sugars was obtained. PMID- 9866173 TI - Repression of the expression of genes encoding xylanolytic enzymes in Aspergillus oryzae by introduction of multiple copies of the xynF1 promoter. AB - A xylanase gene, xynF1, was cloned and characterized from a shoyu koji mould Aspergillus oryzae KBN616. The xynF1 gene was found to be comprised of 1484 bp with ten introns. The deduced amino acid sequence encodes a protein consisting of 327 amino acids (35,402 Da) which is very similar to the fungal family F xylanases such as Aspergillus nidulans XlnC, Aspergillus kawachii XynA and Penicillium chrysogenum XylP. The intron/exon organization of xynF1 is very similar to that of the fungal family F xylanase genes. Plasmid pXPR64, which contains 64 copies of the xynF1 promoter region (PxynF1) in the same direction, was constructed and introduced into A. oryzae. This led to reduced expression of both xylanase and beta-xylosidase genes in the transformants. PMID- 9866174 TI - Electroporation of, plasmid isolation from and plasmid conservation in Clostridium acetobutylicum DSM 792. AB - Procedures have been developed allowing recombinant DNA work with Clostridium acetobutylicum DSM 792. Electroporation was used to introduce plasmid DNA into exponentially growing clostridial cells and 6 x 10(2) transformants/microgram DNA could be obtained at a time constant of 5.5 ms, 1.8 kV, 50 microF, and 600 omega. The method also allowed the taxonomic group IV strain NI-4082 to be transformed (10(1) transformants/microgram DNA). Plasmid preparation from recombinant clostridia was optimal when a modification of the alkaline lysis method was employed. It was also important to use cells from the mid-logarithmic growth phase. Recombinant strains could be easily preserved as spore suspensions; under all conditions tested plasmids were maintained. PMID- 9866175 TI - Isolation and characterization of Enterobacter cloacae capable of metabolizing asparagine. AB - A gram-negative, rod-shaped bacterium capable of utilizing L-asparagine as its sole source of carbon and nitrogen was isolated from soil and identified as Enterobacter cloacae. An intracellularly expressed L-asparaginase was detected and it deaminated L-asparagine to aspartic acid and ammonia. High-pressure liquid chromatography analysis of a cell-free asparaginase reaction mixture indicated that 2.8 mM L-asparagine was hydrolyzed to 2.2 and 2.8 mM aspartic acid and ammonia, respectively, within 20 min of incubation. High asparaginase activity was found in cells cultured on L-fructose, D-galactose, saccharose, or maltose, and in cells cultured on L-asparagine as the sole nitrogen source. The pH and temperature optimum of L-asparaginase was 8.5 and 37-42 degrees C, respectively. The half-life of the enzyme at 30 degrees C and 37 degrees C was 10 and 8 h, respectively. PMID- 9866176 TI - Carbon catabolite repression of invertase during batch cultivations of Saccharomyces cerevisiae: the role of glucose, fructose, and mannose. AB - When Saccharomyces cerevisiae are grown on a mixture of glucose and another fermentable sugar such as sucrose, maltose or galactose, the metabolism is diauxic, i.e. glucose is metabolized first, whereas the other sugars are metabolized when glucose is exhausted. This phenomenon is a consequence of glucose repression, or more generally, catabolite repression. Besides glucose, the hexoses fructose and mannose are generally also believed to trigger catabolite repression. In this study, batch fermentations of S. cerevisiae in mixtures of sucrose and either glucose, fructose or mannose were performed. It was found that the utilization of sucrose is inhibited by concentrations of either glucose or fructose higher than 5 g/l, and thus that glucose and fructose are equally capable of exerting catabolite repression. However, sucrose was found to be hydrolyzed to glucose and fructose, even when the mannose concentration was as high as 17 g/l, indicating, that mannose is not a repressing sugar. It is suggested that the capability to trigger catabolite repression is connected to hexokinase PII, which is involved in the in vivo phosphorylation of glucose and fructose. PMID- 9866177 TI - The influence of carbon source on the level and composition of ceramides of the Candida lipolytica yeast. AB - Candida lipolytica yeast was grown batchwise on two different carbon sources, glucose and n-hexadecane. Free ceramides were quantitatively isolated from sphingolipid fractions of total lipids by a combination of column chromatography and preparative thin-layer chromatography. Their composition, after acid methanolysis, was analysed by gas-liquid chromatography. The ceramide content accounted for 2.6% of the total cell lipids in hexadecane-grown cells, which was 1.5 times higher than in glucose-grown cells. The fatty acid composition of ceramides was characterized by the predominance of fatty acids shorter than 20 carbon atoms and by high concentrations of fatty acids with 16 carbon atoms after growth on both carbon sources. The dominant fatty acid was hydroxylated 16:0 in the glucose-grown cells and 16:0 in the hexadecane-grown cells. The striking finding was the low degree of fatty acid hydroxylation and relatively high proportion of odd-numbered fatty acids in ceramide of the n-hexadecane-grown cells. The ceramides contained an unusual long-chain base composition. In hexadecane-grown cells more than 60% of the long-chain bases were C19 phytosphingosine. In glucose-grown cells more than one-half of the total long chain bases were tetrahydroxy bases, 4,5-dihydroxysphinganine and 4,5 dihydroxyeicosasphinganine. PMID- 9866178 TI - Determination of erythema-effective solar radiation in Japan and Germany with a spore monolayer film optimized for the detection of UVB and UVA--results of a field campaign. AB - The available physical and biological broad-band radiometers designed to determine erythema-effective radiation do not show any response or over/underestimate the biologically effective radiation to a high extent in the ultraviolet (UV)A spectral region. The data presented in this paper demonstrate that the biological system used in this study is the first one to make possible measurements of erythema-effective radiation in the sun in the UVA and UVB spectral region. These measurements were performed with a spore-film filter system as well as with spectroradiometers. It was demonstrated that this biotechnological method could be used to determine exact values expressed as minimal erythemal dose (MED). The spore-film system was tested in various field campaigns performed in Germany and in Japan. The seasonal daily variation of UV radiation in Germany determined in the period November 1995 to December 1996 using the spore-film filter system in sunny conditions tallied well with model calculations. The daily dose in Germany measured with the spore-film system close to the summer solstice, in sunny conditions (20.45 MED), was approximately 20 times higher than the lowest value measured close to the winter solstice (0.82 MED), a result which was in accordance with model calculations. The data determined with the spore-film filter system in Sapporo and Naha, Japan, fitted to the erythema-weighted data calculated from spectroradiometric measurements (Brewer), even at low solar radiation angles in a solar spectrum with less UVB but significant UVA. The spore-film dosimeter values were about 103 +/- 8% of the integrated dose of the Brewer instrument. The standard deviation of the spore film measurements obtained in Japan was 12.8%. The responsivity of the spore-film system towards longer wavelengths within the UVA spectrum was tested with the Okasaki Large Spectrograph with monochromatic radiation. At a wavelength of 365 nm--in a spectral region which is dominant in many tanning lamps and with minor importance for solar radiation in summer conditions--the tested spore-film system gave results that were close (112% compared to the calibration dose) to the calibration dose which was used for irradiation. PMID- 9866179 TI - Organic halogen removal from chlorinated humic ground water and lake water by nitrifying fluidized-bed biomass characterised by electron microscopy and molecular methods. AB - The dechlorinating and genotoxicity-removing activities of nitrifying fluidized bed reactor biomass towards chlorinated organic compounds in water were shown at level below 1 ppm. The removal rates of adsorbable organic halogens were 200 micrograms Cl (g VS day)-1 for chlorinated humic ground water and 50 micrograms Cl (g VS day)-1 for chlorinated lake water when studied in batch mode. In a sequenced batch mode the removal rates [micrograms Cl (g VS day)-1] were 2000 from chlorohumus, 1400-1800 from chlorophenols in chlorinated ground water, and 430-720 from chlorohumus in chlorinated lake water. Genotoxicity was removed to a large extent (60%-80%) from the chlorinated waters upon incubation with nitrifying reactor biomass. 2,6-Di-, 2,4,6-tri and 2,3,4,6-tetrachlorophenols competed with chlorinated water organohalogens for dechlorination. The dechlorination of chlorophenols and chlorohumus required no ammonia and was not prevented by inhibitors of ammonia oxidation, nitrapyrin, parathion, sodium diethyldithiocarbamate, or allylthiourea. Electron microscopical inspection of the biomass showed the dominance of clusters of bacteria resembling known nitrifying species, Nitrosomonas, Nitrobacter, and Nitrosospira. This was supported by polymerase chain reaction amplification of the biomass DNA with four different primers, revealing the presence of 16S rDNA sequences assignable to the same species. The most intensive band obtained with the Nitroso4E primer was shown to be closely related to Nitrosomonas europaea by restriction analysis. PMID- 9866180 TI - Approach and avoidance strength during goal attainment: regulatory focus and the "goal looms larger" effect. AB - Three studies tested the hypothesis that regulatory focus, both as a chronic person variable and as a manipulated situational variable, differentially affects the strength of participants' approach and avoidance strategic motivations as they get closer to the goal. In 2 studies, flexion and extension arm pressure were used as on-line measures of approach and avoidance intensity, respectively. As predicted, the approach gradient was steeper for participants with a promotion focus on aspirations and gains than for participants with a prevention focus on responsibilities and nonlosses, whereas the reverse was true for the avoidance gradient. In a third study, the same pattern of results was found on a persistence measure of motivational strength. Participants with a promotion focus worked longer on anagrams closer to the goal when they were approach means for goal attainment than when they were avoidance means, whereas the reverse was true for participants with a prevention focus. PMID- 9866182 TI - Unraveling social categorization in the "who said what?" paradigm. AB - A multinomial model of the "Who said what?" paradigm (S. E. Taylor, S. T. Fiske, N. J. Etcoff, & A. J. Ruderman, 1978) explains the pattern of participants' assignment errors by means of the joint operation of several processes. Specifically, memory for discussion statements, person memory, category memory, and 3 different guessing processes can be accommodated by the model. The model's ability to disentangle these processes is validated in a series of 5 experiments. The model thereby enables a more refined use of the "Who said what?" paradigm in testing theories of social categorization. This is demonstrated in a 6th experiment in which the validated model is applied to the study of the effects of cognitive load on categorization. PMID- 9866181 TI - The actor as context for social judgments: effects of prior impressions and stereotypes. AB - Three experiments identified conditions under which trait judgments made about a behavior were more likely to influence later judgments of the behavior. In Experiment 1, participants made trait judgments about numerous behaviors presented with photos of actors. Some behaviors were repeated, paired with the same or a different actor. All repeated behaviors were judged faster than new behaviors. Facilitation was greatest when repeated behaviors were paired with the same actor, suggesting greater influence of prior judgments in this condition. Experiments 2 and 3 replicated this effect, and the pattern of response times (RTs) suggested a stronger association between the actor and behavior when a prior impression of the actor had been formed (Experiment 2) and when the behavior was stereotypic of the actor's group (Experiment 3). Level of prejudice moderated RT patterns in Experiment 3. Implications for context effects, the nature of trait inferences, and stereotype change are discussed. PMID- 9866183 TI - Social loafing under fatigue. AB - In 2 experiments, 64 male students worked almost continuously for 20 hr without sleep under varying social conditions. In Experiment 1, participants worked either individually or as a group. As hypothesized, performance deteriorated over time, especially in the group condition, which allowed participants to loaf. In Experiment 2, all participants worked in groups. They were instructed that public feedback would be provided either on the group result only or on the individual results of all group members. As expected, when individual results were made public, performance deteriorated less. Overall, the data suggest that fatigue increases social loafing. However, both individualizing the task and providing public individual feedback seem to counteract these effects. PMID- 9866184 TI - The trouble with friendly faces: skilled performance with a supportive audience. AB - In 3 experiments, supportive audiences were associated with unexpected performance decrements (i.e., "choking" under pressure). On a difficult, skill based task, participants were more likely to fail when observed by supportive audiences than when observed by nonsupportive audiences. When the criterion for success was easy, supportive audiences had no effect. With a difficult criterion, supportive audiences elicited cautious, protective strategies that were associated with poor performance: Speed decreased without improving accuracy. Despite impairments caused by supportive audiences, performers found supportive audiences more helpful and less stressful than neutral or adversarial audiences, and participants believed (wrongly) that they performed better with a supportive audience. Results suggest that people are not aware of debilitating effects of supportive audiences and may opt for emotional comfort rather than objective success. PMID- 9866185 TI - Teasing in hierarchical and intimate relations. AB - Following E. Goffman's (1967) face threat analysis of social interaction, it was hypothesized that the aggressive, playful content of teasing would vary according to social status and relational satisfaction, personality, role as teaser or target, and gender. These 4 hypotheses were tested in analyses of the teasing among fraternity members (Study 1) and romantic couples (Study 2). Consistent with a face threat analysis of teasing, low-status fraternity members and satisfied romantic partners teased in more prosocial ways, defined by reduced face threat and increased redressive action. Some findings indicate that disagreeable individuals teased in less prosocial ways, consistent with studies of bullying. Targets reported more negative emotion than teasers. Although female and male romantic partners teased each other in similar ways, women found being the target of teasing more aversive, consistent with previous speculation. PMID- 9866186 TI - The immunological effects of thought suppression. AB - Individuals often suppress emotional thoughts, particularly thoughts that arouse negative emotions, as a way of regulating mood and reducing distress. However, recent work has highlighted the complexities and unexpected cognitive and physiological effects of thought suppression. In a study designed to examine the short-term immunological effects of thought suppression, participants wrote about either emotional or nonemotional topics with or without thought suppression. Blood was drawn before and after each experimental session on 3 consecutive days. Results showed a significant increase in circulating total lymphocytes and CD4 (helper) T lymphocyte levels in the emotional writing groups. Thought suppression resulted in a significant decrease in CD3 T lymphocyte levels. The implications of the results for the role of the expression and suppression of emotion in health are discussed. PMID- 9866187 TI - Life events and social support as moderators of individual differences in cardiovascular and cortisol reactivity. AB - Whether prior stress increases acute stress reactivity is unresolved. The impact of life events (within the past 12 months) and social support on cardiovascular responses was investigated in 90 young male firefighters. Cardiovascular and cortisol measures were collected across baseline, arithmetic, and speech tasks; intertask recovery; and three recovery trials. Reactivity differences were not independently associated with life events. High social support was associated with greater arithmetic cardiovascular reactivity and faster recovery. Combined life events and social support grouping showed that effects of support were accentuated when event frequency was high, suggesting that life events and support interacted to sensitize future stressor responses. Support may promote the alerting response mobilization but prevent chronic allostatic load by enhancing recovery. PMID- 9866188 TI - Avoidance personal goals and the personality-illness relationship. AB - Three studies demonstrated that avoidance personal goals are positively related to physical symptom reports. These results were obtained (a) using both longitudinal and retrospective methodologies and (b) controlling for neuroticism and other alternative predictor variables. In 2 of the studies, a process model was validated in which perceived competence and perceived controlledness were shown to mediate the observed relationship between avoidance goals and symptomatology. Specifically, avoidance goals predicted perceived competence and perceived controlledness, and these variables in turn predicted longitudinal and retrospective symptom reports. Ancillary results help clarify the unique roles of neuroticism and avoidance goals as predictors of physical symptomatology. PMID- 9866189 TI - Bright, bad, babyfaced boys: appearance stereotypes do not always yield self fulfilling prophecy effects. AB - Three studies tested the hypothesis that babyfaced adolescent boys would compensate for the undesirable expectation that they will exhibit childlike traits by behaving contrary to it. Studies 1 and 2 revealed that babyfaced boys from middle- and lower class samples, including a sample of delinquents, showed higher academic achievement than their mature-faced peers, refuting the stereotype of babyfaced people as intellectually weak. In the lower class samples, this compensation effect was moderated by IQ and socioeconomic status (SES), variables that influence the ability to overcome low expectations. Study 3 showed that babyfaceness also can produce negative compensatory behaviors. Low SES babyfaced boys were more likely than their mature-faced peers to be delinquent, and babyfaced delinquents committed more crimes, refuting the stereotype of babyfaced people as warm, submissive, and physically weak. PMID- 9866190 TI - Temperament, recalled parenting styles, and self-regulation: testing the developmental postulates of self-discrepancy theory. AB - Self-discrepancy theory (SDT) postulates that self-regulatory systems corresponding to the ideal and ought self-domains emerge from the influences of temperament (e.g., sensitivity to stimuli for positive vs. negative outcomes) and socialization (e.g., parenting behaviors and interpersonal outcome contingencies). This article reports 2 studies testing the developmental postulates of SDT concurrently and retrospectively. Study 1 showed that self regulation with reference to the ideal vs. the ought domain was differentially associated with recollections of parenting styles of warmth and rejection, respectively. In Study 2, these findings were replicated, and self-regulation with reference to the ideal vs. ought domain was discriminantly associated with questionnaire measures of positive vs. negative temperament. Findings support the developmental postulates of SDT, despite the limitations of retrospective studies. PMID- 9866191 TI - The effect of age on positive and negative affect: a developmental perspective on happiness. AB - The effect of age on happiness, as defined by positive and negative affect, was examined in a survey of 2,727 persons of a broad age range (25-74) conducted by the MacArthur Foundation Research Network on Successful Midlife Development. The age-affect association was examined, controlling for a host of sociodemographic, personality, and contextual influences. Among women, age was related to positive affect nonlinearly but was unrelated to negative affect. Among men, age interacted with 2 key variables in predicting affect: extraversion and marital status. These findings lend support to recent life span theories of emotion and indicate that personality, contextual, and sociodemographic variables, as well as their interactions, are all needed to fully understand the age-affect relationship. PMID- 9866192 TI - Gender differences on negative affectivity: an IRT study of differential item functioning on the Multidimensional Personality Questionnaire Stress Reaction Scale. AB - Item response theory methods were used to study differential item functioning (DIF) between gender groups on a measure of stress reaction. Results revealed that women were more likely to endorse items describing emotional vulnerability and sensitivity, whereas men were more likely to endorse items describing tension, irritability, and being easily upset. Item factor analysis yielded 5 correlated factors, and the DIF analysis, in turn, revealed differential gender mean differences on these factors. This finding illustrates how even in an essentially unidimensional scale, comparison of group mean differences can be affected by multidimensionality caused by item clusters that share similar content. Results do not support arguments that measures of negative affective dispositions "artificially" produce gender mean differences by focusing on specific selected content areas. PMID- 9866193 TI - Psychological distress and the frequency of perfectionistic thinking. AB - Five studies tested the hypotheses that there are individual differences in the frequency of automatic thoughts involving perfectionism and that these thoughts are associated with psychological distress. Research with the Perfectionism Cognitions Inventory (PCI) established that this new measure has adequate psychometric properties, and high PCI scorers tend to spontaneously report perfectionistic thoughts in naturalistic situations. Additional research confirmed that frequent perfectionism thoughts account for unique variance in distress, over and above variance predicted by standard measures of negative automatic thoughts and trait perfectionism measures. Overall, the findings support the view that personality traits involved in depression and anxiety have a cognitive component involving ruminative thoughts and that activation of this cognitive personality component contributes to distress. PMID- 9866194 TI - HLA-H: a pseudogene with increased variation due to balancing selection at neighboring loci. AB - The HLA complex includes the most polymorphic genes in the human genome. However, the HLA class Ib loci have little, if any, nucleotide variation, presumably due to their specialized functions or perhaps due to a lack of function. This population genetic study of HLA-H, a class I pseudogene, was initiated to determine the pattern of variation at neutral sites within the HLA complex. We found that the pattern of variation for HLA-H is consistent with the neutral model. However, the amount of variation in HLA-H is significantly greater than estimates for other silent sites within the human genome outside of the MHC (theta = 0.0144, P < 0.000001). Our study further indicates that other possible causes of increased variation such as a high mutation rate for HLA-H, interlocus gene conversion, increased diversity in the sample population in general, and selection acting directly on HLA-H are unlikely. Instead, these data suggest that HLA-H has increased variation as a result of balancing selection acting on nearby loci such as HLA-A. PMID- 9866195 TI - Distribution of the retrotransposable element 412 in Drosophila species. AB - Copy numbers of sequences homologous to the Drosophila melanogaster retrotransposable element 412, their distribution between the chromosome arms and the chromocenter, and whether they contain full-size copies were analyzed for 55 species of the Drosophila genus. Element 412 insertion sites were detected on the chromosome arms of D. melanogaster, Drosophila simulans, and a few species of the obscura group, but the chromocenter was labeled in almost all species. The presence of element 412 sequences in the majority of species shows that this element has a long evolutionary history in Drosophilidae, although it may have recently invaded the chromosomes in some species, such as D. simulans. Differences in copy number between species may be due to population size or specific endogenous or environmental factors and may follow the worldwide invasion of the species. Putative full-length copies were detected in the chromocenters of some species with no copies on the chromosome arms, suggesting that the chromocenter may be a shelter for such copies and not only for deleted ones. PMID- 9866196 TI - Models of amino acid substitution and applications to mitochondrial protein evolution. AB - Models of amino acid substitution were developed and compared using maximum likelihood. Two kinds of models are considered. "Empirical" models do not explicitly consider factors that shape protein evolution, but attempt to summarize the substitution pattern from large quantities of real data. "Mechanistic" models are formulated at the codon level and separate mutational biases at the nucleotide level from selective constraints at the amino acid level. They account for features of sequence evolution, such as transition transversion bias and base or codon frequency biases, and make use of physicochemical distances between amino acids to specify nonsynonymous substitution rates. A general approach is presented that transforms a Markov model of codon substitution into a model of amino acid replacement. Protein sequences from the entire mitochondrial genomes of 20 mammalian species were analyzed using different models. The mechanistic models were found to fit the data better than empirical models derived from large databases. Both the mutational distance between amino acids (determined by the genetic code and mutational biases such as the transition-transversion bias) and the physicochemical distance are found to have strong effects on amino acid substitution rates. A significant proportion of amino acid substitutions appeared to have involved more than one codon position, indicating that nucleotide substitutions at neighboring sites may be correlated. Rates of amino acid substitution were found to be highly variable among sites. PMID- 9866197 TI - The origin and differentiation of the heteromorphic sex chromosomes Z, W, X, and Y in the frog Rana rugosa, inferred from the sequences of a sex-linked gene, ADP/ATP translocase. AB - Sex chromosomes of the Japanese frog Rana rugosa are heteromorphic in the male (XX/XY) or in the female (ZZ/ZW) in two geographic forms, whereas they are still homomorphic in both sexes in two other forms (Hiroshima and Isehara types). To make clear the origin and differentiation mechanisms of the heteromorphic sex chromosomes, we isolated a sex-linked gene, ADP/ATP translocase, and constructed a phylogenetic tree of the genes derived from the sex chromosomes. The tree shows that the Hiroshima gene diverges first, and the rest form two clusters: one includes the Y and Z genes and the other includes the X, W, and Isehara genes. The Hiroshima gene shares more sequence similarity with the Y and Z genes than with the X, W, and Isehara genes. This suggests that the Y and Z sex chromosomes originate from the Hiroshima type, whereas the X and W chromosomes originate from the Isehara-type sex chromosome. Thus, we infer that hybridization between two ancestral forms, with the Hiroshima-type sex chromosome in one and the Isehara type sex chromosome in the other, was the primary event causing differentiation of the heteromorphic sex chromosomes. PMID- 9866198 TI - Microsatellite variation in Drosophila melanogaster and Drosophila simulans: a reciprocal test of the ascertainment bias hypothesis. AB - Interspecific comparisons of microsatellite loci have repeatedly shown that the loci are longer and more variable in the species from which they are derived (the focal species) than are homologous loci in other (nonfocal) species. There is debate as to whether this is due to directional evolution or to an ascertainment bias during the cloning and locus selection processes. This study tests these hypotheses by performing a reciprocal study. Eighteen perfect dinucleotide microsatellite loci identified from a Drosophila simulans library screen and 18 previously identified in an identical Drosophila melanogaster library screen were used to survey natural populations of each species. No difference between focal and nonfocal species was observed for mean PCR fragment length. However, heterozygosity and number of alleles were significantly higher in the focal species than in the nonfocal species. The most common allele in the Zimbabwe population of both species was sequenced for 31 of the 36 loci. The length of the longest stretch of perfect repeat units is, on average, longer in the focal species than in the non-focal species. There is a positive correlation between the length of the longest stretch of perfect repeats and heterozygosity. The difference in heterozygosity can thus be explained by a reduction in the length of the longest stretch of perfect repeats in the nonfocal species. Furthermore, flanking-sequence length difference was noted between the two species at 58% of the loci sequenced. These data do not support the predictions of the directional evolution hypothesis; however, consistent with the ascertainment bias hypothesis, the lower variability in nonfocal species is an artifact of the microsatellite cloning and isolation process. Our results also suggest that the magnitude of ascertainment bias for repeat unit length is a function of the microsatellite size distribution in the genomes of different species. PMID- 9866199 TI - The complete mitochondrial DNA sequence of the shark Mustelus manazo: evaluating rooting contradictions to living bony vertebrates. AB - A remarkable example of a misleading mitochondrial protein tree is presented, involving ray-finned fishes, coelacanths, lungfishes, and tetrapods, with sea lampreys as an outgroup. In previous molecular phylogenetic studies on the origin of tetrapods, ray-finned fishes have been assumed as an outgroup to the tetrapod/lungfish/coelacanth clade, an assumption supported by morphological evidence. Standard methods of molecular phylogenetics applied to the protein encoding genes of mitochondria, however, give a bizarre tree in which lamprey groups with lungfish and, therefore, ray-finned fishes are not the outgroup to a tetrapod/lungfish/coelacanth clade. All of the dozens of published phylogenetic methods, including every possible modification to maximum likelihood known to us (such as inclusion of site heterogeneity and exclusion of potentially misleading hydrophobic amino acids), fail to place the ray-finned fishes in a biologically acceptable position. A likely cause of this failure may be the use of an inappropriate outgroup. Accordingly, we have determined the complete mitochondrial DNA sequence from the shark, Mustelus manazo, which we have used as an alternative and more proximal outgroup than the lamprey. Using sharks as the outgroup, lungfish appear to be the closest living relative of tetrapods, although the possibility of a lungfish/coelacanth clade being the sister group of tetrapods cannot be excluded. PMID- 9866200 TI - Estimating the rate of evolution of the rate of molecular evolution. AB - A simple model for the evolution of the rate of molecular evolution is presented. With a Bayesian approach, this model can serve as the basis for estimating dates of important evolutionary events even in the absence of the assumption of constant rates among evolutionary lineages. The method can be used in conjunction with any of the widely used models for nucleotide substitution or amino acid replacement. It is illustrated by analyzing a data set of rbcL protein sequences. PMID- 9866201 TI - Granule-bound starch synthase: structure, function, and phylogenetic utility. AB - Interest in the use of low-copy nuclear genes for phylogenetic analyses of plants has grown rapidly, because highly repetitive genes such as those commonly used are limited in number. Furthermore, because low-copy genes are subject to different evolutionary processes than are plastid genes or highly repetitive nuclear markers, they provide a valuable source of independent phylogenetic evidence. The gene for granule-bound starch synthase (GBSSI or waxy) exists in a single copy in nearly all plants examined so far. Our study of GBSSI had three parts: (1) Amino acid sequences were compared across a broad taxonomic range, including grasses, four dicotyledons, and the microbial homologs of GBSSI. Inferred structural information was used to aid in the alignment of these very divergent sequences. The informed alignments highlight amino acids that are conserved across all sequences, and demonstrate that structural motifs can be highly conserved in spite of marked divergence in amino acid sequence. (2) Maximum-likelihood (ML) analyses were used to examine exon sequence evolution throughout grasses. Differences in probabilities among substitution types and marked among-site rate variation contributed to the observed pattern of variation. Of the parameters examined in our set of likelihood models, the inclusion of among-site rate variation following a gamma distribution caused the greatest improvement in likelihood score. (3) We performed cladistic parsimony analyses of GBSSI sequences throughout grasses, within tribes, and within genera to examine the phylogenetic utility of the gene. Introns provide useful information among very closely related species, but quickly become difficult to align among more divergent taxa. Exons are variable enough to provide extensive resolution within the family, but with low bootstrap support. The combined results of amino acid sequence comparisons, maximum-likelihood analyses, and phylogenetic studies underscore factors that might affect phylogenetic reconstruction. In this case, accommodation of the variable rate of evolution among sites might be the first step in maximizing the phylogenetic utility of GBSSI. PMID- 9866203 TI - Escherichia coli molecular phylogeny using the incongruence length difference test. AB - Molecular phylogeny of the species Escherichia coli using the E. coli reference (ECOR) collection strains has been hampered by (1) the absence of rooting in the commonly used phenogram obtained from multilocus enzyme electrophoresis (MLEE) data and (2) the existence of recombination events between strains that scramble phylogenetic trees reconstructed from the nucleotide sequences of genes. We attempted to determine the phylogeny for E. coli based on the ECOR strain data by extracting from GenBank the nucleotide sequences of 11 chromosomal structural and 2 plasmid genes for which the Salmonella enterica homologous gene sequences were available. For each of the 13 DNA data sets studied, incongruence with a nonnucleotide whole-genome data set including MLEE, random amplified polymorphic DNA, and rrn restriction fragment length polymorphism data was measured using the incongruence length difference (ILD) test of Farris et al. As previously reported, the incongruence observed between the gnd and plasmid gene data and the whole-genome data was multiple, indicating numerous horizontal transfer and/or recombination events. In five cases, the incongruence detected by the ILD test was punctual, and the donor group was identified. Congruence was not rejected for the remaining data sets. The strains responsible for incongruences with the whole genome data set were removed, leading to a "prior-agreement" approach, i.e., the determination of a phylogeny for E. coli based on several genes, excluding (1) the genes with multiple incongruences with the whole genome data, (2) the strains responsible for punctual incongruences, and (3) the genes incongruent with each other. The obtained phylogeny shows that the most basal group of E. coli strains is the B2 group rather than the A group, as generally thought. The D group then emerges as the sister group of the rest. Finally, the A and B1 groups are sister groups. Interestingly, the most primitive taxon within E. coli in terms of branching pattern, i.e., the B2 group, includes highly virulent extraintestinal strains with derived characters (extraintestinal virulence determinants) occurring on its own branch. PMID- 9866202 TI - Cytosolic isocitrate dehydrogenase in humans, mice, and voles and phylogenetic analysis of the enzyme family. AB - In this study, we report cDNA sequences of the cytosolic NADP-dependent isocitrate dehydrogenase for humans, mice, and two species of voles (Microtus mexicanus and Microtus ochrogaster). Inferred amino acid sequences from these taxa display a high level of amino acid sequence conservation, comparable to that of myosin beta heavy chain, and share known structural features. A Caenorhabditis elegans enzyme that was previously identified as a protein similar to isocitrate dehydrogenase is most likely the NADP-dependent cytosolic isocitrate dehydrogenase enzyme equivalent, based on amino acid similarity to mammalian enzymes and phylogenetic analysis. We also suggest that NADP-dependent isocitrate dehydrogenases characterized from alfalfa, soybean, and eucalyptus are most likely cytosolic enzymes. The phylogenetic tree of various isocitrate dehydrogenases from eukaryotic sources revealed that independent gene duplications may have given rise to the cytosolic and mitochondrial forms of NADP dependent isocitrate dehydrogenase in animals and fungi. There appears to be no statistical support for a hypothesis that the mitochondrial and cytosolic forms of the enzyme are orthologous in these groups. A possible scenario of the evolution of NADP-dependent isocitrate dehydrogenases is proposed. PMID- 9866204 TI - B30.2-like domain proteins: update and new insights into a rapidly expanding family of proteins. AB - The B30.2 domain is a conserved region of around 170 amino acids associated with several different protein domains, including the immunoglobulin folds of butyrophilin and the RING finger domain of ret finger protein. We recently reported several novel members of this family as well as previously undescribed protein families possessing the B30.2 domain. Many proteins have subsequently been found to possess this domain, including pyrin/marenostrin and the midline 1 (MID1) protein. Mutations in the B30.2 domain of pyrin/marenostrin are implicated in familial Mediterranean fever, and partial loss of the B30.2 domain of MID1 is responsible for Opitz G/BBB syndrome, characterized by developmental midline defects. In this study, we scrutinized the available sequence data bases for the identification of novel B30.2 domain proteins using highly sensitive database searching tools. In addition, we discuss the chromosomal localization of genes in the B30.2 family, since the encoded proteins are likely to be involved in other forms of periodic fever, autoimmune, and genetic diseases. PMID- 9866205 TI - 28S and 18S rDNA sequences support the monophyly of lampreys and hagfishes. AB - Resolving the interrelationships of three major extant lineages of vertebrates (hagfishes, lampreys, and gnathostomes) is a particularly important issue in evolution, because the basal resolution critically influences our understanding of primitive vertebrate characters. A consensus has emerged over the last 20 years that lampreys are the sister group to the gnathostomes and the hagfishes represent an ancient, basal lineage. This hypothesis has essentially displaced the classical hypothesis of monophyly of the cyclostomes (lampreys plus hagfishes). To test these hypotheses, we compared nearly complete ribosomal DNA sequences from each of these major lineages, as well as those from a cephalochordate and a urochordate, which represent a paraphyletic outgroup for assessing the basal vertebrate relationships. For this comparison, 92%-99% complete 28S rDNA sequences were obtained from the lancelet Branchiostoma floridae, the hagfish Eptatretus stouti, the lamprey Petromyzon marinus, and cartilaginous fishes Hydrolagus colliei and Squalus acanthias and were then analyzed with previously reported 28S and 18S rDNA sequences from other chordates. We conducted conventional (nonparametric) bootstrap analyses, under maximum-likelihood, parsimony, and minimum-evolution (using LogDet distances) criteria, of both 28S and 18S rDNA sequences considered separately and combined. All these analyses provide moderate to very strong support for the monophyly of the cyclostomes. Furthermore, the currently accepted hypothesis of a lamprey gnathostome clade is moderately rejected by the Kishino-Hasegawa test (P = 0.099) and resoundingly rejected by parametric bootstrap tests (P < 0.01) in favor of monophyly of living cyclostomes. Another significant finding is that the hagfish E. stouti has the longest 28S rDNA gene known in any organism (> 5,200 nt). PMID- 9866206 TI - Substitution bias, rapid saturation, and the use of mtDNA for nematode systematics. AB - Only relatively recently have researchers turned to molecular methods for nematode phylogeny reconstruction. Thus, we lack the extensive literature on evolutionary patterns and phylogenetic usefulness of different DNA regions for nematodes that exists for other taxa. Here, we examine the usefulness of mtDNA for nematode phylogeny reconstruction and provide data that can be used for a priori character weighting or for parameter specification in models of sequence evolution. We estimated the substitution pattern for the mitochondrial ND4 gene from intraspecific comparisons in four species of parasitic nematodes from the family Trichostrongylidae (38-50 sequences per species). The resulting pattern suggests a strong mutational bias toward A and T, and a lower transition/transversion ratio than is typically observed in other taxa. We also present information on the relative rates of substitution at first, second, and third codon positions and on relative rates of saturation of different types of substitutions in comparisons ranging from intraspecific to interordinal. Silent sites saturate extremely quickly, presumably owing to the substitution bias and, perhaps, to an accelerated mutation rate. Results emphasize the importance of using only the most closely related sequences in order to infer patterns of substitution accurately for nematodes or for other taxa having strongly composition-biased DNA. ND4 also shows high amino acid polymorphism at both the intra- and interspecific levels, and in higher level comparisons, there is evidence of saturation at variable amino acid sites. In general, we recommend using mtDNA coding genes only for phylogenetics of relatively closely related nematode species and, even then, using only nonsynonymous substitutions and the more conserved mitochondrial genes (e.g., cytochrome oxidases). On the other hand, the high substitution rate in genes such as ND4 should make them excellent for population genetics studies, identifying cryptic species, and resolving relationships among closely related congeners when other markers show insufficient variation. PMID- 9866207 TI - Hierarchical analysis of variation in the mitochondrial 16S rRNA gene among Hymenoptera. AB - Nucleotide sequences from a 434-bp region of the 16S rRNA gene were analyzed for 65 taxa of Hymenoptera (ants, bees, wasps, parasitoid wasps, sawflies) to examine the patterns of variation within the gene fragment and the taxonomic levels for which it shows maximum utility in phylogeny estimation. A hierarchical approach was adopted in the study through comparison of levels of sequence variation among taxa at different taxonomic levels. As previously reported for many holometabolous insects, the 16S data reported here for Hymenoptera are highly AT rich and exhibit strong site-to-site variation in substitution rate. More precise estimates of the shape parameter (alpha) of the gamma distribution and the proportion of invariant sites were obtained in this study by employing a reference phylogeny and utilizing maximum-likelihood estimation. The effectiveness of this approach to recovering expected phylogenies of selected hymenopteran taxa has been tested against the use of maximum parsimony. This study finds that the 16S gene is most informative for phylogenetic analysis at two different levels: among closely related species or populations, and among tribes, subfamilies, and families. Maximization of the phylogenetic signal extracted from the 16S gene at higher taxonomic levels may require consideration of the base composition bias and the site-to-site rate variation in a maximum likelihood framework. PMID- 9866208 TI - Y chromosome variation of mice and men. AB - DNA sequences from the nonrecombining portion of the Y chromosome were compared with autosomal and X-linked sequences from mice and humans to test the neutral prediction that ratios of polymorphism to divergence are the same for different genes. Intraspecific variation within Mus domesticus was compared with divergence between M. domesticus and Mus caroli for Sry, a region 5' to Sry, and four X linked genes, Hprt, Plp, Amg, and Glra2. None of these comparisons revealed significantly reduced variation on the Y chromosome. Intraspecific variation within humans was compared with divergence between humans and chimpanzees for three Y-linked loci (Zfy, the YAP region, and the Sry region), seven X-linked loci (Il2rg, Plp, Hprt, Gk, Ids, Pdhal, and Dmd), and the beta-globin locus on chromosome 11. In these comparisons, the observed level of variation on the human Y chromosome was slightly lower than expected, but was significantly lower in only one case (Sry region vs. Dmd). These results suggest that the levels of variability on the Y chromosome in mice and humans are close to expected values given the effective population size and mutation rates for these loci. There is at most only a modest reduction in variability that may be attributed to natural selection (either genetic hitchhiking or background selection). PMID- 9866209 TI - The mutation rates of di-, tri- and tetranucleotide repeats in Drosophila melanogaster. AB - In a recent study, we reported that the combined average mutation rate of 10 di-, 6 tri-, and 8 tetranucleotide repeats in Drosophila melanogaster was 6.3 x 10(-6) mutations per locus per generation, a rate substantially below that of microsatellite repeat units in mammals studied to date (range = 10(-2)-10(-5) per locus per generation). To obtain a more precise estimate of mutation rate for dinucleotide repeat motifs alone, we assayed 39 new dinucleotide repeat microsatellite loci in the mutation accumulation lines from our earlier study. Our estimate of mutation rate for a total of 49 dinucleotide repeats is 9.3 x 10( 6) per locus per generation, only slightly higher than the estimate from our earlier study. We also estimated the relative difference in microsatellite mutation rate among di-, tri-, and tetranucleotide repeats in the genome of D. melanogaster using a method based on population variation, and we found that tri- and tetranucleotide repeats mutate at rates 6.4 and 8.4 times slower than that of dinucleotide repeats, respectively. The slower mutation rates of tri- and tetranucleotide repeats appear to be associated with a relatively short repeat unit length of these repeat motifs in the genome of D. melanogaster. A positive correlation between repeat unit length and allelic variation suggests that mutation rate increases as the repeat unit lengths of microsatellites increase. PMID- 9866210 TI - The molecular evolution of sperm bindin in six species of sea urchins (Echinoida: Strongylocentrotidae). AB - The acrosomal protein bindin attaches sperm to eggs during sea urchin fertilization. Complementary to ongoing functional biochemical studies, I take a comparative approach to explore the molecular evolution of bindin in a group of closely related free-spawning echinoid species. Two alleles of the mature bindin gene were sequenced for each of six species in the sea urchin family Strongylocentrotidae. The nucleotide sequences diverged by at least 1% per Myr at both silent and replacement sites. Two short sections flanking the conserved block show an excess of nonsynonymous substitutions. Each is homologous to a region that had been identified as a target of selection in other sea urchin comparisons. A large proportion of the bindin-coding sequence consists of a highly variable repeat region. Bindin sequences, even including the large intron, could not resolve the branching order among five of the species. PMID- 9866211 TI - Mitochondrial gene order is not conserved in arthropods: prostriate and metastriate tick mitochondrial genomes. AB - The entire mitochondrial genome was sequenced in a prostriate tick, Ixodes hexagonus, and a metastriate tick, Rhipicephalus sanguineus. Both genomes encode 22 tRNAs, 13 proteins, and two ribosomal RNAs. Prostriate ticks are basal members of Ixodidae and have the same gene order as Limulus polyphemus. In contrast, in R. sanguineus, a block of genes encoding NADH dehydrogenase subunit 1 (ND1), tRNA(Leu)(UUR), tRNA(Leu)(CUN), 16S rDNA, tRNA(Val), 12S rDNA, the control region, and the tRNA(Ile) and tRNA(Gln) have translocated to a position between the tRNA(Glu) and tRNA(Phe) genes. The tRNA(Cys) gene has translocated between the control region and the tRNA(Met) gene, and the tRNA(Leu)(CUN) gene has translocated between the tRNA(Ser)(UCN) gene and the control region. Furthermore, the control region is duplicated, and both copies undergo concerted evolution. Primers that flank these rearrangements confirm that this gene order is conserved in all metastriate ticks examined. Correspondence analysis of amino acid and codon use in the two ticks and in nine other arthropod mitochondrial genomes indicate a strong bias in R. sanguineus towards amino acids encoded by AT-rich codons. PMID- 9866212 TI - Swimmer 1, a new low-copy-number LINE family in teleost genomes with sequence similarity to mammalian L1. PMID- 9866214 TI - [Levels of the pineal hormone melatonin and its circadian variations in the cochlea of Wistar rats]. AB - The normal inner ear contains different elements belonging to the immune system. Some pathology of the inner ear has been attributed to autoimmune mechanisms, both humoral and cellular. Melatonin, the principal hormone of the pineal gland, modulates the immune system, is involved in autoimmune processes, and mediates free-radical and interleukine production. In a study of the presence of melatonin in rat cochlea, a day-night rhythm of melatonin in cochlea was observed. Melatonin levels are high during dark periods and low during light periods. This rhythm is synchronic with peripheral melatonin circulation. More work, both experimental and clinical, is needed to determine the role of melatonin in the inner ear and its therapeutic usefulness. PMID- 9866213 TI - [Nitric oxide synthase activity in nasal mucosa]. AB - Nitric oxide (NO) plays an important regulatory role in airway function and seems to be implicated in the pathophysiology of several airway diseases. We studied the presence of NO synthase activity in human nasal mucosa and nasal polyp tissues obtained from patients undergoing septoplasty or polypectomy, respectively. NO synthase activity was quantified in tissue homogenates using citrulline release assay and was located in tissue sections using NADPH diaphorase histochemistry. The results indicated that nasal polyps contain higher levels of total NO synthase activity than nasal mucosa tissue. In addition, nasal polyps contained mainly inducible NO synthase activity whereas all NO synthase activity detected in the nasal mucosa was in constitutive form. In both cases, NO synthase activity was localized in epithelial cells. In view of these results, we conclude that NO may be an important inflammatory mediator in the respiratory system and that the epithelium may be a source of NO production. PMID- 9866215 TI - [Microsurgical approach and graft anastomosis to the trunk of the facial nerve in the Wistar rat]. AB - An experimental model was developed for the purpose of investigating neural regeneration of the facial nerve in Wistar rats. The anatomic references used to locate the facial nerve at its exit from the skull base are described. A detailed description of the microsurgical anastomosis of cryopreserved nerve allografts is given. The major advantages and disadvantages of rat models compared with other animal models for experimental studies of the facial nerve are discussed. PMID- 9866216 TI - [Gold weights for the treatment of lagophthalmos caused by facial paralysis. Our experience and review of the literature]. AB - Paralysis of the orbicularis oculi muscle in patients with facial palsy can originate serious functional and esthetic problems. The implantation of a gold weight in the upper eyelid is one of the many surgical techniques that have been described for the correction of lagophthalmos in these patients. In this article we review the literature and present our experience with gold weight implantation for eyelid reanimation in patients with facial palsy. This procedure is technically simple to perform, produces good functional and cosmetic results, has a relatively low complication rate and is reversible should facial function return. PMID- 9866217 TI - [Hearing in premature infants. Evoked-cochlear microphones]. AB - This report summarizes our experience in obtaining microphonic potentials in human beings. Special studies were made to obtain microphonic potentials in premature newborns. PMID- 9866218 TI - [Naso-alveolar cysts. A report of 10 new cases and a review of the literature]. AB - Nasoalveolar cysts, also known as Klestadt's or nasolabial cysts, are uncommon midfacial lesions. Ten cases treated at the 12 de Octubre University Hospital are reported, describing the most relevant clinical and pathological findings are reported. The main theories of the origin of these developmental, non-odontogenic cysts are discussed. A review of the literature is included. PMID- 9866219 TI - [The incidence and progression of lymphoma of the cavum in AIDS]. AB - A clinical case of a woman with AIDS who developed lymphoma of the nasopharynx and paralysis of the VII, IX and XII cranial nerves is reported. The probable origin of the paralysis in relation to AIDS and lymphoma is discussed. PMID- 9866220 TI - [Tonsillectomy by electrical dissection versus blunt dissection: a study of 838 cases (Randomized trial of 207 cases and non-randomized prospective study of 631 cases]. AB - Tonsillectomy is one of the most common operations performed in the world and various techniques and instruments have evolved for this purpose. We report the results of two prospective studies: in the first study, blunt dissection was compared with electrodissection tonsillectomy (207 cases), and in the second study, 631 patients had their tonsils removed using only the electrodissection technique. Although electrodissection tonsillectomy is not common in Spain, in our experience it is simple, reliable and easy to learn. Its advantages include minimal bleeding, shorter operating time, and less postoperative hemorrhage. The postoperative morbidity is similar to that of blunt dissection. PMID- 9866221 TI - [Radiotherapy alone versus neo-adjuvant chemotherapy and irradiation in the treatment of carcinoma of the cavum]. AB - Radiation therapy is the usual treatment for nasopharyngeal carcinoma. However, in recent years the use of neoadyuvant chemotherapy in the treatment of local and regionally advanced carcinoma has been investigated. We report the results of a retrospective study of two treatments used in our center. The study included 68 patients: 34 (group A) who received radiotherapy alone and 34 (group B) who received neoadyuvant chemotherapy before radiotherapy. In group A, 70.6% achieved a complete clinical response: 23.5% relapsed: 5 patients presented distant metastases. Survival rates at 5 and 10 years were 53% and 27% respectively; the disease-free survival was 71.4% at 5 years and 54% at 10 years. In group B, the complete clinical response rate to neoadyuvant chemotherapy was 35.3%, which increased to 73.5% when the treatment was complemented with radiotherapy. The relapse rate was 14.7%; the survival rates at 5 and 10 years were 49.5% and 49%, respectively; and the disease-free survival was 77.2% at 5 and 10 years. PMID- 9866222 TI - [Current results of the treatment of cervical metastatic lymph nodes by concurrent hyperfractionation of carboplatin and irradiation]. AB - The results of the treatment of metastatic neck nodes is evaluated after a mean follow-up of 24 months (maximum 45 months). Fifty-seven patients with epidermoid carcinoma of the head and neck were treated according to a hyperfractionated chemoradiation schedule including two fractions a day. Each fraction consisted of 10 mg carboplatin + 115 cGy. Two fractions were given each day, five days a week, for a total dose of 700 mg carboplatin + 8050 cGy. Whenever possible, surgical salvage was performed if treated nodes persisted or recurred. Ten patients presented with N0, 8 with N1, 7 with N2a, 4 with N2b, 7 with N2c, and 21 with N3. The classification of the primary tumor was: 3 Tx, 6 T2, 9 T3 and 39 T4. One hundred and eleven nodes were treated (62 with a diameter of 1-3 cm, 26 with a diameter of 3-6 cm and 23 with a diameter over 6 cm). Actuarial node controls were: 100% for N0, 97% for nodes 1-3 cm, 87% for nodes 3-6 cm, 95% for nodes over 6 cm and 97% for the whole group. The actuarial local-regional control was 71% and the disease-free survival was 60%. These results include 5 surgical salvages (11% of N+), 2 of which recurred again (40%), while another 3 (60%) did not recur. PMID- 9866223 TI - [Bilateral peripheral facial paralysis and Guillain-Barre syndrome]. AB - The Guillain-Barre syndrome is a polyneuropathy of acute onset that initially tends to produce motor damage of the lower limbs, albumin-cytological dissociation in cerebrospinal fluid and electrophysiological findings suggestive of demyelination. This entity produces a wide variety of symptoms, including damage to the facial nerve in as many as half of all episodes, which means that this syndrome should be considered in the differential diagnosis of facial palsy, particularly bilateral and synchronous cases. In a review of six cases of Guillain-Barre syndrome in which bilateral facial palsy occurred at some point, a history of immediate infection was confirmed in five, mainly related to Herpesvirinae and Campylobacter. One case began with bilateral palsy, whereas palsy appeared almost simultaneously with the rest of the symptoms in the other cases. High protein levels without cells in cerebrospinal fluid and electrophysiological patterns of slow facial nerve conduction were confirmed in every patient. Treatment with intravenous immunoglobulins resolved all episodes in less than six months. We conclude by reiterating the need for awareness of this syndrome in every case of peripheral VIIth nerve palsy, especially bilateral cases, although the associated symptoms were not very useful, if at all useful. PMID- 9866224 TI - [Differential features of transglottic carcinoma]. AB - A study was made of the differential features of transglottic laryngeal carcinomas. We made a retrospective study of patients with laryngeal carcinoma diagnosed in our center from 1985 to 1996. In this period, 1212 laryngeal carcinomas were diagnosed, of which 99 (8%) were considered transglottic. In relation to factors such as sex, toxic habits, level of histological differentiation or affected nodes, patients with transglottic carcinomas had characteristics that were intermediate between patients with glottic and supraglottic carcinomas. An analysis was made of the treatments and the results obtained in patients treated with radical intention and a minimum follow-up of 3 years. The treatment most frequently used in patients with transglottic carcinoma was surgery (total laryngectomy) and complementary radiotherapy. The 5-year actuarial adjusted survival rate jor patients with transglottic carcinoma was 78%, which was intermediate between the survival rates of patients with glottic (90%) or supraglottic (69%) carcinoma. PMID- 9866226 TI - [Auricular pseudocyst]. AB - Two cases of auricular pseudocyst in men are presented. In both cases, the pseudocyst was located on the anterior surface of the upper part of the outer ear. Conservative treatment with repeated aspirations in one case, and incision and drainage in the other, were curative and produced good esthetic results. PMID- 9866225 TI - [Subcutaneous sinusoid cavernous hemangioma of the outer ear. Review of literature]. AB - Hemangionas of the ear have been rarely described. A few cases have been reported in the world literature involving the middle ear space or tympanic membrane, and a few more involving the inner ear. Cavernous hemangioma often is associated with other hemangiomas, and there is almost always a cutaneous segment that facilitates the diagnosis. The authors report a rare case (to their knowledge, the first to be described) of a subcutaneous cavernous hemangioma of the auricular concha. The diagnosis and treatment are discussed and the literature is reviewed. PMID- 9866227 TI - [Cerebral abscess as a complication of chronic otitis media]. AB - A case is reported of chronic otitis media and cholesteatoma in an adult patient with behavioral disorders who developed a pyogenic brain abscess. Initially, clinical management focused on the otogenic disease in view of the patient's symptoms and medical history. Finally, less easily recognized discriminating signs and symptoms, and the results of imaging studies were emphasized. These data confirmed the clinical diagnosis of pyogenic brain abscess secondary to cholesteatoma. The case illustrates how one of the most severe infective complications related with cholesteatoma may appear and its a remainder to clinicians to take this possibility into consideration. PMID- 9866228 TI - [Pharyngeal tuberculosis: a clinical case]. AB - A case of pharyngeal tuberculosis secondary to pulmonary tuberculosis is reported. The importance of biopsy for the histopathological and microbiological diagnosis is emphasized. PMID- 9866229 TI - [Primary tuberculosis of the lingual tonsil. A case report]. AB - A case of primary tuberculosis of the lingual amygdala is reported. This is a highly unusual location in the head and neck area. This entity can be easily misdiagnosed as a result of confusion with other pathologies. PMID- 9866230 TI - [MALT lymphoma of the parotid gland associated with Sjogren syndrome. A case report and review of literature]. AB - Sjogren syndrome (SS) is a chronic inflammatory process involving the exocrine glands. Its association with the development of lymphomas is well documented, and this risk has estimated to be 44 times that of the general population. A MALT lymphoma of the parotid gland in a patient with Sjogren syndrome is reported. We review the literature and comment the clinical and histopathological features of these tumors. PMID- 9866231 TI - Leadership in the managed care era: challenges, conflict, ambivalence. AB - Mental health organizations are in the midst of massive changes brought about by managed care. This paper explores attitudes and reactions of leaders in mental health organizations to these changes. The leader's own conflicts regarding this approach to mental health care is a major determinant of why some organizations adapt more readily than others. Strategies for leaders that recognize the need to address conflict within the individual and the organization are discussed. The importance of facilitating dynamic interactional processes between leaders and non-leaders in regard to mutual expectations also is emphasized. PMID- 9866232 TI - The effect of provider characteristics on case management activities. AB - The authors investigated the impact of provider characteristics on the performance of clinical case managers. Case managers in a large California county were interviewed using a structured instrument that assessed service linkage, assertive outreach, and family management. Findings indicate that assertive outreach was done more often by more experienced case managers, and less often by those with larger caseloads. Belief in the efficacy of an activity was associated with doing it much more frequently. PMID- 9866233 TI - Practice patterns and hospitalization rates. AB - Basic indicators of community support program treatment outcome as well as the degree to which the programs conform to practice guidelines for major depression are evaluated. Hospitalization rates subsequent to treatment are measured, and the correlation between hospitalization rates and practice patterns is determined. Data sets that describe outpatient and inpatient services over a 4 year period, but do not include common person identifiers were analyzed using probabilistic population estimates. Results indicate that there is substantial variation among the community programs in practice patterns and hospitalization rates, and the two are negatively correlated. PMID- 9866234 TI - The quality of VA mental health services. AB - This study examines whether male and female veterans differ on either subjective or objective measures of the quality of VA mental health care. The study sample were all discharged with a psychiatric diagnosis from a VA inpatient mental health program. Results indicate that women were less satisfied with inpatient care, but were mixed on their satisfaction with overall mental health care. Males and females did not differ on the quality of inpatient care, but women had significantly higher quality of outpatient care. These results not only highlight the need to stratify or adjust quality measures by gender, but also highlight the potential confounding effect of sub-group specific health behaviors on measures of quality. PMID- 9866235 TI - Franklin County individualized care case rate project. PMID- 9866236 TI - Double jeopardy: the challenge of providing mental health services to older persons. PMID- 9866237 TI - [Analgesic agents: comparative evaluation, mechanisms of action, prospects]. AB - A classification of analgesics based on the localization and mechanisms of their action is proposed. Opioid analgesics are compared: opioid receptor agonists, agonists-antagonists, and partial agonists. Central-action nonopioid analgesics are listed: alpha 2-adrenomimetics, Na- and K-channel blockers, reverse monoamine neuronal capture inhibitors, stimulating amino acid antagonists, etc. Peripheral action analgesics (cycloxygenase inhibitors) are described. PMID- 9866238 TI - [Antinociceptive components of general anesthesia and postoperative analgesia]. AB - The philosophy of full-value antinociceptive protection of patients during and after surgical interventions is discussed. The author presents her viewpoint on the problem with due consideration for recent published reports. She advocates a prophylactic approach to attaining the antinociceptive protection (the so-called "preemptive analgesia"). Analgesics of peripheral action are the main means for protection at the peripheral level. Peripheral antialgogens contrycal (trasilol), acelysin (aspisol), baralgine, diclofenac, and ketorolac were effectively used during the pre-, intra-, and postoperative periods: the course of total anesthesia and postoperative period improved, intensity of the painful syndrome and the need in opioids was decreased, and wound healing improved. For preventing secondary (central) hyperalgesia and decreasing the intensity of postoperative painful syndrome, NMDA-receptor antagonist ketamine in microdoses is recommended before and during surgery. Afferent nociceptive flow can be decreased by local and regional anesthesia. Practical application of the preemptive analgesia principle improves the efficacy and safety of analgesia at different stages of surgical treatment, decreases total and local (at the level of operated tissues) consequences of surgical trauma, and decreases the probability of complications. PMID- 9866239 TI - [Use of H-reflex in the assessment of the excitability of spinal cord nociceptive neurons in man]. PMID- 9866240 TI - [Study of mechanisms of action of amitriptyline and acupuncture using nociceptive flexor reflex in patients with chronic forms of headache]. AB - The nociceptive flexor reflex (NFR, R3) was tried for quantitative assessment of pain in patients with various forms of primary and secondary headaches. Amitriptyline and acupuncture elevated the threshold of R3-reflex emergence, though the threshold of subjective pain sensitivity increased only in response to amitriptyline. NFR is adequate for assessing anesthesia efficacy and investigating the mechanisms of action of analgesics in patients with headache. PMID- 9866242 TI - [Comparative evaluation of the use of nalbuphine and buprenorphine in prehospital care]. AB - Analgesic efficacy of nalbufine and buprenorphine is assessed in 86 patients with painful syndrome caused by unstable angina and acute myocardial infarction and in 72 patients with locomotor injuries, to whom urgent care was rendered by ambulance teams before hospitalization. By the velocity and depth of analgesic effect nalbufine is not inferior to morphine, and in patients with unstable angina and myocardial infarction is even superior to it. High analgesic activity of buprenorphine is compatible to that of morphine, but the rate of analgesia development is insufficient for urgent care. PMID- 9866241 TI - [Role of neuropeptides and "pain substances" in the formation of humoral mechanisms of experimental and postoperative pain]. AB - Humoral mechanisms of pain caused by different factors vary. The authors compare blood concentrations of "painful substances" in experimental dogs and in patients suffering from postoperative pain relieved by electroacupuncture for assessing the role of these substances: serotonin, histamine, prostaglandin F2 alpha (PGF), and neuropeptides beta-endorphine, methionine- and leucine-enkephalines. Serotonin, histamine, and PGF participated in the nociception process in an equal measure both in dogs and humans. Notable differences were observed for neuropeptides, which can be explained by species-specific differences and by probable contribution of other neuropeptides to mechanisms of experimental and postoperative pain. PMID- 9866243 TI - [Interactions of general anesthetics, analgesics and antihypertensive agents]. PMID- 9866244 TI - [Analgesia and treatment of muscular shivering during the period of postanesthetic adaptation in oncologic patients]. PMID- 9866245 TI - [Parenteral use of oruvel (ketoprofen) for analgesia in the postoperative period]. AB - Antiinflammatory and analgesic effects of oruvel (ketoprofene), a nonsteroid antiinflammatory drug, are demonstrated by many scientists. Both intramuscular and oral oruvel is acknowledged as an effective means for therapy of inflammatory and dystrophic rheumatic diseases. Moreover, oruvel in 100-mg flasks for infusions is intended for the treatment of postoperative pain, specifically, in orthopaedic surgery. PMID- 9866246 TI - [Pharmacology of local anesthetics and clinical aspects of segmental blocking. II. Spinal anesthesia]. AB - Clinical picture of development of segmental blocking after subarachnoidal injection of hyperbaric solutions of 0.75% bupivacaine, 5% ultracaine, and isobaric 0.5% bupivacaine is studied. A total of 152 patients operated on the lower part of the body and the lower limbs were examined under conditions of single, prolonged subarachnoidal, and combined spinal epidural anesthesia. Ultracaine and bupivacaine in different concentrations with different barism provided anesthesia equivalent by the efficacy, depth, and dissemination of sensory block. Segmental blocking with 5% ultracaine was characterized by the shortest latent period (3.14 +/- 0.16 min, p < 0.05) but was no shorter (124.1 +/ 3.37 min) than operative analgesia with 0.75% hyperbaric bupivacaine (120.0 +/- 5.10 min). Isobaric bupivacaine provided the longest effective analgesia (215.0 +/- 45.0 min, p < 0.05). Microcatheter technique improved the safety and control of subarachnoidal anesthesia in comparison with a single injection, and combined spinal epidural anesthesia shortened the latent period of segmental blocking and ensured intraoperative anesthesia and postoperative analgesia at the expense of the epidural component. PMID- 9866247 TI - [Prospects of the use of caudal epidural anesthesia]. AB - Caudal epidural anesthesia for interventions on the lower limbs and pelvic organs was used in 525 patients. A specific feature of the method is use of hypoosmolic local anesthetic solution (osmolality 260 mosmol/kg) containing lidocaine, 0.9% sodium chloride, and distilled water. Pathologic studies showed that in adult patients, at least 40 ml anesthetic should be injected into the caudal canal for adequate blocking. During surgery, caudal epidural anesthesia reliably protected from surgical trauma without side effects for respiration and circulation. The duration of analgesic effect was 3 +/- 0.5 h and even longer, if local anesthesia was potentiated with sedative drugs. No complications were observed, failures occurred in 5.2% cases. The method is simple and reliable and is recommended for practice. PMID- 9866248 TI - [Improvement of epidural anesthesia in reconstructive surgery of the aorta in Leriche's syndrome]. AB - Fifty-six patients subjected to aortofemoral bilateral shunting under prolonged epidural anesthesia are examined. Swan-Gans catheter was used to evaluate central and pulmonary hemodynamics. Ganglionic blocking with pentamine for taxiphylaxis was carried out in 31 patients prior to catheterization of the epidural space. Epidural anesthesia with ganglional blocking is associated with reduced vasodilation and hypotension and attenuates the hemodynamic reactions during clamping and unclamping of the aorta, thus decreasing the incidence of intra- and postoperative complications. PMID- 9866249 TI - [Epidural anesthesia using local anesthetics and clopheline in geriatric urology]. AB - Study of pharmacokinetics of lidocaine, trimecaine, and clofelin demonstrated rapid resorption of these drugs in the postoperative period. Comparison of two methods of epidural anesthesia in high-risk patients (ASA 3) subjected to urologic surgery showed the objective advantages of adrenergic regional anesthesia, ensuring the stability of vital functions during surgery and allowing operations in awaken patients with adequate spontaneous respiration. PMID- 9866251 TI - [Our general chronic pain (centers for treatment chronic pain: therapeutic and organizational work principles)]. AB - Management of chronic pain is now not only medical but a very important social and economic problem. The approaches to its management in specialized pain control centers are discussed basing on the 20-year experience of the first such center in Russia: pain control clinic with therapeutic and diagnostic center "Integrative Medicine" at Research Center of Surgery. The problems of personnel for such pain control centers, cost-effect issues, diagnostic and therapeutic methods including an advanced technique, resonance electropuncture analgesia and therapy, are reviewed. PMID- 9866250 TI - [Prolonged epidural analgesia induced by clopheline in combination with lidocaine in obstetric analgesia]. AB - The study was carried out in 178 women without grave obstetrical or extragenital diseases. In group 1 labor pain was relieved by prolonged epidural anesthesia with 2% lidocaine solution (2-2.5 mg/kg), in group 2 prolonged epidural anesthesia with 1% lidocaine solution (1 mg/kg) and 0.01% clofelin (1 microgram/kg) was administered. Central hemodynamics, heart rhythm, external respiration function, uterine contractility, and fetal intrauterine status were assessed. The findings indicate that none of the methods had a negative impact on the vital parameters of women and newborns at any stage of anesthesia. However, a combination of epidural clofelin (1 microgram/kg) with lidocaine permits an appreciable decrease in the doses of both drugs without decreasing the efficacy of anesthesia. This method has a favorable effect on the course of labor: the mouth of the womb opens sooner at a lower uterine activity and there are no negative effects on the fetus and newborn. PMID- 9866252 TI - [Current principles of multidisciplinary treatment of pain in orthopedic clinics]. AB - Differentiated strategy of treating patients with acute and chronic pain is developed. Preemptive analgesia is a priority trend in the treatment of acute postoperative pain. The most prevalent method of postoperative analgesia is prolonged opioid epidural analgesia carried out in intensive care wards and other wards by an acute pain management team. For treating patients with chronic painful syndromes, protocols of initial clinical and diagnostic evaluation are developed, permitting the choice of individual treatment strategy. Differentiated complex drug therapy planned with consideration for individual course of the painful syndrome is the basis of treating patients with phantom pain syndrome. Algorithms of differentiated therapy of radicular and spondylogenic pain are designed. Stage-by-stage analysis of treatment efficacy is carried out using modern electrophysiological methods. Realization of the proposed organization principles improved the efficacy of postoperative analgesia to 88.2%, prevented the development of postoperative painful syndrome in 35.6% cases, decreased the incidence of phantom pain syndrome after amputation of the limb from 63.3 to 31.6% and increased the efficacy of this syndrome treatment to 70.1%, and increased the efficacy of treating vertebrogenic painful syndromes to 82.3%. PMID- 9866253 TI - [Rural environment: a challenge for the future]. PMID- 9866254 TI - [Health and social problems of caregivers of patients with dementia]. AB - OBJECTIVE: To describe the physical and psychological morbidity of carers of patients with dementia and to compare them with a group of carers of non-demented patients in the home care programme. DESIGN: Descriptive crossover study, based on face-to-face interviews, with a reference group for comparison. SETTING: Community, Toledo and Esquivias Health Districts. PARTICIPANTS: 61 carers of patients with dementia, and 61 of patients in home care. MEASUREMENTS AND MAIN RESULTS: The interview collected the following variables: social, demographic, morbidity (chronic pathologies, insomnia, anxiety-depression...), medication and interference with social life. Both samples were homogeneous for age, sex, marital status, relationship to patient and educational level of the carers, and age and sex of the patients. The problems most often referred to by the two groups of carers were osteo-muscular. The most commonly used drugs were analgesic NSAIDs, hypertension and anxiety-depression drugs. Insomnia was significantly more common (p < 0.05) in carers of demented patients (63.93%) than in the other group (44.26%), although consumption of sleeping draughts was similar. The Goldberg test was positive in 49.18% of carers of demented patients and in 31.15% of the others (p < 0.05). Carers of demented patients had a more restricted social life (p < 0.01). Only 25.41% thought that their doctor and/or nurse were concerned about their situation as carer. CONCLUSIONS: We found more anxiety depression and insomnia in carers of demented patients. Interference of care with their social life was greater in this group. Carers view the interest shown in their problems by health-workers as insufficient. PMID- 9866255 TI - [Evaluation of a supervised physical exercise program in primary care]. AB - OBJECTIVE: To compare self-perception of health with physical parameters in a population group with chronic illnesses when conducting a group intervention of supervised physical exercise. DESIGN: Intervention study with test-retest evaluation. SETTING: Serraparera Primary Care Centre (PCC) in the town of Cerdanyola del Valles (24,735 inhabitants). PATIENTS: Patients between 30 and 65 with a chronic illness and attended at the PCC (n = 87). INTERVENTION: A seven month programme, based on regular supervised physical exercise, three times a week for an hour, supplemented by organised monthly walks on varying routes. MEASUREMENTS AND MAIN RESULTS: Blood pressure, BMI, flexibility, sub-maximum and at-rest heart rates were measured, and the Nottingham Health Profile (NHP) self perception test conducted. 87 patients began the programme and 51 reached the end. The following was observed in the 51: self-perception of overall improvement in health on the NHP of 9.51 points (CI 95%, 5.73-13.23); lowering of blood pressure, systolic 4.8 mmHg (CI 2.47-7.13) and diastolic 4.78 (CI 3.16-6.46); sub maximum heart rate, with an average difference of 7.65 pulses per minute; and at rest heart rate, with a difference of 4.61 ppm. Flexibility increased by 1.37 (CI 1.05-1.69). There was no significant reduction in BMI. CONCLUSIONS: Patients with chronic illnesses experienced a notable improvement in the control of their pathology and vital constants. There was also a significant difference detected in patients' perception of their own health. PMID- 9866256 TI - [Physician's perception of mental malaise in a basic health district]. AB - OBJECTIVES: a) To determine the prevalence of psychological malaise (PM) in the population-group attending. b) To find the sensitivity and specificity of diagnoses of anxiety-depression made by doctors and on the Goldberg Anxiety and Depression Scale (GADS), using the Mini International Neuropsychiatric Interview (MINI) as gold standard. c) To study whether there are factors that might affect PM's detection. DESIGN: Crossover study. SETTING: Urban health centre. PARTICIPANTS: 252 patients from the general population between 15 and 65 who attended for consultation for 10 days in May 1997. MEASUREMENTS AND RESULTS: The clinical diagnosis of 1266 patients was recorded. A sample was selected at random, to whom both the GADS and the MINI were administered, so as to compare the diagnoses. The prevalence of PM according to clinical judgement was similar to that found with the MINI (40.9%), with a clear overdiagnosis on the GADS. Clinical judgement mainly detected the anxiety component. The results of clinical judgement and the GADS were compared with the MINI (gold standard), and sensitivity of 73.78% and specificity of 72.48% were found for clinical judgement, and sensitivity of 95% and specificity of 64.4% for the GADS. CONCLUSIONS: Prevalence of PM is high. The GADS showed it was useful for PM screening. The doctor's ability to detect PM is considerable. We found no variables that might affect diagnostic error. PMID- 9866257 TI - [Description of costs and effectiveness of a hypertension control program in primary care]. AB - OBJECTIVE: To find the control degree of a population of hypertension patients. To evaluate the cost effectiveness of health services. SETTING: EAP Salvador Pau. Valencia (Spain). PATIENTS: 200 patients diagnosed of hypertension included in the health center file. MEASUREMENTS AND RESULTS: Items studied were age, sex, diagnosis and program inclusion year, number of visits, other explorations and its cost, TAS and TAD, drugs and its cost, personnel cost. We established different well control levels and we obtained the cost of each of them. Twenty six point five per cent of patients had TA values below 140/90 mmHg; Sixty four point five per cent had values below 160/90 mmHg. And sixty six point nine had values below 90 mmHg. Monthly medium cost per patient was 3242 pts. Monthly medium cost per patient well controlled was 12234 pts. CONCLUSIONS: Only a third of our patients are well controlled according the current standards. Efficiency decreases noteworthy when control levels are lower. PMID- 9866258 TI - [Determinants of clinical decision making in primary care: opinion of physicians]. AB - OBJECTIVE: The effective dissemination of scientific information and its adoption by clinicians is the desired outcome of research and the path towards evidence based clinical practice. However, the link between both ends, is not direct and not always attained. Many other determinants play decisive roles in clinical decision making. Their nature, relationship and relative importance are not well understood. DESIGN: We conducted a survey among primary care physicians to ascertain their perception on factors related to clinical decision making. The questionnaire was specifically developed and was administered in face to face interviews by trained interviewers. PARTICIPANTS: A multistage clustered sample of 800 physicians representative of all Spanish primary care physicians, was drawn out of Health Service Payroll lists. Frequencies and multivariate logistic regressions were performed using SUDAAN 7.0. RESULTS: Spanish physicians seem to pay special importance to organizational aspects, such as daily demand, administrative regulations or prescription costs. Expert opinion and textbooks appear as the principal source of information, systematic literature reviews are unknown for most physicians. Finally, the majority of them perceive the need for more education. CONCLUSIONS: Primary care physicians perceive organizational limitations to their decision making process and are willing to improve their medical knowledge. Additionally, they may benefit from strategies towards the spread of evidence based medicine built on these results, such as using experts to effectively convey evidence based information, increasing opportunities for more education, or developing evidence based sources following textbooks advantages. PMID- 9866259 TI - [User satisfaction in primary care]. AB - OBJECTIVES: To find user satisfaction and assess the usefulness of an analogue scale in measuring satisfaction. DESIGN: Multi-centre, descriptive and crossover study. SETTING: Four primary care centres in Girona. PARTICIPANTS: All those users who happened to attend on a certain day (n = 1349). MEASUREMENTS AND MAIN RESULTS: A multi-dimensional, self-administered and anonymous 33-item questionnaire on satisfaction, which had already been validated in primary care, was used. An analogue scale, running from 0 to 10, on overall satisfaction was added. The mean age was 48.5; 33% were men, 62% women. 84.2% replied; 37.3% needed help to reply. Mean satisfaction on the questionnaire was 85%, with lower ratings given to organisational questions, and the highest to personal treatment. The analogue scale achieved 7.34 average. The correlation between the two was r = 0.63 (p < 0.001). CONCLUSIONS: The satisfaction level was similar to that found in other studies. The analogue scale correlated closely with the questionnaire, but was quicker and more simple. As such, it could be used to monitor user satisfaction along with open questions for a supplementary qualitative evaluation. PMID- 9866260 TI - [Diagnosis of maternal and infant health in Palestine]. AB - OBJECTIVE: To describe the health situation of the target population (women between 15 and 45 and children from 0 to 3), living in the towns on the West Bank in which the 12 dispensaries of the Palestine NGO Health Services Council were located, in order to plan a programme of support activities for that NGO. DESIGN: Crossover survey. Descriptive analysis of the data. SETTING: 12 towns in the 5 districts of the West Bank (Palestine) between May and July 1994. PARTICIPANTS: Women of a fertile age (15 to 45) and children between 0 and 3 living in the towns where the 12 dispensaries of the NGO Health Services Council were located. MEASUREMENTS AND MAIN RESULTS: Social, demographic and health variables of the population group involved, the availability of primary care services and the pattern of health service use were studied. Worthy of mention were the average fertility of fertile women of 4.55 deliveries (CI 95%, 4.30-4.80), the use of family planning methods by 61% of the women surveyed, the low indices of mother and infant malnutrition, and the frequency of episodes of acute respiratory infections and diarrhoea in the under-3s. The availability of health services in these communities is wide. The non-governmental and private Palestinian services were preferred for antenatal control, whereas the Israeli government services were preferred for deliveries, as these services had more resources. CONCLUSIONS: The findings were typical of an intermediately developed society, similar to neighbouring countries. The multiplicity of health services available and their lack of coordination could make the application of a unitary health policy in the area difficult. PMID- 9866262 TI - [Original article. III. Results]. PMID- 9866261 TI - [New etiopathogenic aspects of obesity]. PMID- 9866263 TI - [Drugs and maternal lactation. Inadequate information in the Vade Mecum]. PMID- 9866264 TI - [Informative abstracts and publication standards. Are the standards only for the authors?]. PMID- 9866265 TI - [Has scarlet fever returned or did it ever go away?]. PMID- 9866266 TI - [Should we continue to lower arterial blood pressure with calcium antagonists?]. PMID- 9866267 TI - [Do our patients wish to receive hormone replacement therapy during menopause? Myths that persist]. PMID- 9866268 TI - [Tropical medicine]. PMID- 9866269 TI - [Indicators of anxiety and depression in subjects with different kinds of diet: vegetarians and omnivores]. AB - The following study, one of the first done in Puerto Rico, investigate the different kinds of diet and the level of anxiety and depression that the subjects present. The sample consists of 80 subjects between 25 and 70 years age divided into two main groups (vegetarian versus no vegetarian) depending their diet consumption. The basic findings in the three psychological tests given (IDARE-1, IDARE-2 and CES-D) to the subjects demonstrate significant differences in anxiety and depression between groups. More anxiety and depression where reported in the no vegetarian groups in comparison with the vegetarian groups. In addition, diet analysis found more nutritional antioxidant agents levels in the vegetarian group in comparison with the no-vegetarian group. PMID- 9866270 TI - Gastroschisis: a ten year review. AB - From 1983 to 1993, 30 cases of gastroschisis were managed at the Mayaguez Medical Center. Ninety percent of these patients underwent primary closure of their abdominal wall defect. Three of 30 patients (10%) required silastic or goretex silos with final closure in an average of 8 days. There was no sex predilection, the average birth weight was 2.4 kg and the mean gestational age was 36 weeks. Thirty percent had associated anomalies, the majority were intestinal atresia, and/or undescended testicles. Twenty one (70%) of infants were delivered vaginally. Nine children (30%) were delivered via cesarean section. Four cesarean sections were done solely after prenatal ultrasonic identification of gastroschisis. There was no improvement in hospital stay, complications, or days until enteral feeds were tolerated when vaginally delivered patients were compared to those born by c-sections. In seven patients mesh sheeting (Marlex) was used for closure of late hernia defects. The mean hospital stay was 50 days and the mean time to enteral feedings 20 days. All patients required postoperative mechanical ventilation for an average of 4 days. There was no mortality. Our data and review of the literature do not support gastroschisis prenatal diagnosis as a sole indication for cesarean section. Our data showed favorable prognosis for most babies. Primary fascial closure can be accomplished safely in the majority of patients. No single operative strategy is ideal for all patients, and treatment of individual defects should be tailored to the degree of visceroabdominal disproportion. PMID- 9866271 TI - Pediatric thyroid nodules: insights in management. AB - BACKGROUND: Multiple diagnostic studies are utilized to unveil malignancy in pediatric thyroid nodules and determine whether surgical therapy is needed. PURPOSE: The aim of this report was to determine whether management of pediatric thyroid nodules has changed with the current use of diagnostic modalities such as ultrasonography (US), radionuclear scans (RNS) and fine needle aspiration biopsy (FNAB). MATERIAL/METHODS: Twenty-four children with thyroid nodules managed during a ten-year period comprised the study group. Demographic characteristics, clinical manifestations, US and RNS imaging findings, FNAB results, surgical therapy, complications and pathological reports were retrospectively reviewed. US, RNS and FNAB results were categorized as either benign, malignant, suspicious or insufficient. RESULTS: Females outnumbered males by a five to one ratio. Mean age was 14.9 years. Nineteen nodules were benign (79%) and five malignant (21%). All children were euthyroid. Benign nodules were soft, movable, solitary and nontender. Malignant nodules were characterized by localized tenderness, a multiglandular appearance, and fixation to adjacent tissues. US and RNS gave no clue toward management since cystic and hot nodules figured among malignant cases respectively. US achieved 86% accuracy, 80% sensitivity and 88% specificity; RNS showed 26% accuracy, 80% sensitivity and 11% specificity; FNAB achieved 80% accuracy, 60% sensitivity and 90% specificity. Suppressive thyroid hormone therapy was useless in the few cases tried. Physical examination findings, persistence of the nodule, progressive growth and cosmetic appearance where the most common indications for surgery. CONCLUSIONS: Present diagnostic modalities played a minor role in the decision to withhold surgery. US was useful for aiming aspiration of cystic nodules. RNS decided the functionality of the nodule, but its accuracy was far from ideal. FNAB is a safe procedure whose greatest help was to resolve in case of suspicious or malignant cytology that a more radical procedure is needed. Clinical judgement as determined by serial physical findings and suspicion continues to be the most determinant factors in the management of thyroid nodules in children. PMID- 9866272 TI - Primitive peripheral neuroectodermal tumors Mayaguez Medical Center experience. AB - Primitive neuroectodermal or neuroepithelial tumors are names used to describe neoplasias composed of undifferentiated cells resembling germinal cells of the embryonic neural tube. These tumors are small round cell malignancies of the neural crest origin arising outside the central and sympathetic nervous system. They are described as peripheral and central neuroectodermal tumors related to the original malignant cell. A great number of tumors are described under this classification in spite of the fact that there is no an universal acceptance that these small-cell neoplasms, regardless of their primary site, are derived from immature neuroectoderm tissue. Because one tumor resembles others in terms of its phenotypic expression, multiple specific studies such as clinical profile, ultrastructural, immunocytochemical, and cytogenetic features should be studied, since no single clinical or laboratory marker is by itself diagnostic. However, there is a chromosomal reciprocal translocation, t(11;22)(q24;q12), which is unique to Primitive Neuroectodermal Tumor (PNET). PMID- 9866273 TI - Uretero-arterial fistula: a case report and review of the literature. AB - We report the case of a uretero-arterial fistula (UAF) formation in a 68 years old male who had previously undergone an Aortobifemoral graft. He got complicated with occlusion and infection of the right lower extremity requiring a right hip disarticulation for its management. This was followed by groin infection and graft protusion, managed by transabdominal resection of the right graft limb, at which time the right ureter was lacerated and repaired. Several months later, he presented with gross hematuria found to be secondary to UAF. The diagnostic and management steps leading to this patient care will be reviewed, together with a review of the literature pertinent to this case report. PMID- 9866274 TI - Recognition of hemophilia A in an elderly patient. AB - Hemophilia A (classic hemophilia) is an hereditary coagulation disorder characterized by the absence, severe deficiency, or defective functioning of plasma coagulation factor VIII. It is inherited in an X-linked recessive manner and occurs almost exclusively in males. The first manifestations of bleeding are usually first noted as a young child since most of the patients with hemophilia A have a profound deficiency of factor VIII (less than 1% of normal value). However, in mild hemophilia (5-25% of normal level of factor VIII) the condition may escape detection with many of the patients developing bleeding only after trauma or surgery. Hemophilia A is the result of a recent genetic mutation in approximately one third of patients, for whom often there is no family history of a bleeding disorder. Here we present an elderly male patient with spontaneous bleeding in an extremity that has coagulation studies consistent with the diagnosis of hemophilia A. Physicians must be aware that mild hemophilia can present in this unusual manner and should consider this possibility in patients that have unexplained bleeding even if there is no clear personal or family history of an hereditary coagulation disorder. PMID- 9866276 TI - Solid and papillary neoplasm of the pancreas: a case presentation. AB - Solid and papillary tumors of the pancreas are very rare malignancies, more commonly occurring in young women. They usually present as asymptomatic, large abdominal masses, and different from the most common neoplasm of the pancreas, which is the adenocarcinoma, these tumors have a high percentage of curability when treated by complete surgical resection. (6) For this reason, when the diagnosis of Frantz's tumor is made or strongly suspected, every attempt should be made for complete surgical excision since curability is high and radiotherapy and, or chemotherapy are of no use for its treatment. PMID- 9866275 TI - Sudden periodic paralysis: rare manifestation of thyrotoxicosis. AB - Nonfamilial hypokalemic thyrotoxic periodic paralysis is rarely diagnosed among Caucasians and blacks in the western world but it is relatively common among Asiatics. Sudden paralysis occurring while at rest after a large carbohydrate meal or strenuous exercise in an undiagnosed mild thyrotoxic patient is a common presentation. A case illustrating such presentation is reported. Intracellular shifts of potassium triggered or facilitated by hyperthyroidism and hyperinsulinemia are the biochemical features. Correction of the thyrotoxic state is the definitive treatment for this disorder. Judicious administration of potassium is indicated during the hypokalemic episode to prevent life-threatening arrhythmias. PMID- 9866278 TI - Acute spinal cord and head injury: case report and discussion of cardiac, respiratory and endocrine abnormalities. AB - We report a male patient who after a fall suffered high cervical spinal cord and head (cerebral) injuries. These injuries led to spinal shock, marked sinus bradycardia and asystolic cardiac and respiratory arrests, recalcitrant central traumatic diabetes insipidus, and death within approximately seven weeks. Temporary transvenous cardiac pacing proved useful in the management of this patient. PMID- 9866277 TI - Bifid scrotum, perineal hamartoma and high imperforate anus: a case report. AB - This is a case report of a newborn patient with imperforate anus, urethro-colonic fistula, perianal hamartoma, and bifid scrotum. Successful staged repair of these anomalies is described together with review of the embriology related to the case. PMID- 9866279 TI - Infectious diseases in England and Wales: January to March 1998. PMID- 9866280 TI - Vero cytotoxin producing Escherichia coli O157--redesignation of strains of phage types 21 and 28. PMID- 9866281 TI - Inadvertent administration of BCG to schoolchildren. PMID- 9866282 TI - PHLS Group B Streptococcus Working Group. PMID- 9866283 TI - Crisis in nursing academics is not brewing--it is present. PMID- 9866284 TI - National attention to nursing education is needed. PMID- 9866285 TI - Unbundling research, teaching, and practice is wrong solution. PMID- 9866286 TI - Some clinical faculty are indentured servants. PMID- 9866287 TI - Holistic doctoral education for holistic nursing. PMID- 9866288 TI - Too little discussion of tobacco-related diseases and deaths. PMID- 9866289 TI - Nursing's first Senior Scholar at the U.S. Agency for Health Care Policy and Research. Interview by Peter I Buerhaus. PMID- 9866290 TI - Nurse staffing levels and adverse events following surgery in U.S. hospitals. AB - PURPOSE: To examine the relationship between nurse staffing and selected adverse events hypothesized to be sensitive to nursing care, while controlling for related hospital characteristics. Efforts in the United States to reduce hospital costs, resulting in strategies to use fewer nurses, have stimulated extensive debate but little evaluation. DESIGN: Survey using data from a 20% stratified probability sample to approximate U.S. community hospitals. The sample included 589 acute-care hospitals in 10 states. METHODS: Discharge data from 1993 for patients aged 18 years and over were used to create hospital-level adverse event indicators. These hospital-level data were matched to American Hospital Association data on community hospital characteristics, including nurse staffing, to examine the relationship between nurse staffing and adverse events. RESULTS: A large and significant inverse relationship was found between full-time-equivalent RNs per adjusted inpatient day (RNAPD) and urinary tract infections after major surgery (p < .0001) as well as pneumonia after major surgery (p < .001). A significant but less robust inverse relationship was found between RNAPD and thrombosis after major surgery (p < .01), as well as pulmonary compromise after major surgery (p < .05). CONCLUSIONS: Inverse relationships between nurse staffing and these adverse events provide information for managers to use when redesigning and restructuring the clinical workforce employed in providing inpatient care. PMID- 9866291 TI - Use of immigration policy to manage nursing shortages. AB - PURPOSE: To examine long-term implications of using temporary, nonimmigrant nurse programs to manage fluctuations in the demand for registered nurses. ORGANIZING FRAMEWORK: This discussion is located in the full context of migration--reviewing theories and concepts of labor migration--referring to experience with guest worker programs worldwide, outlining recent nursing shortages in the United States, describing the Immigration Nursing Relief Act (INRA), and raising questions for nurses in the United States and in the global marketplace. SOURCES: Review of scholarly literature on international migration, existing studies on nurse migration to the United States, and original research, conducted between 1992 and 1994, for the Immigration Nursing Relief Advisory Committee (INRAC) Report. METHODS: Policy analysis of theories, concepts, and perspectives related to nurse migration. FINDINGS: In the United States, highly skilled foreign nurses tend to complement rather than displace local labor. Yet recruiting foreign educated nurses for entry-level jobs perpetuates patterns of dependency in the sending country and delays creative solutions to staff development in the host country. Nonimmigrant status creates a vulnerable workforce. There may be a disparity between the ideal of nurse migration as collaborative exchange and the reality of institutionalized occupational migration networks. CONCLUSIONS: While foreign nurse recruitment might solve short-term needs, repetitive temporary nurse migration programs create long-term consequences that are not in the best interests of the profession. The absence of consistent policy creates an opportunity for nursing to take an active role in developing the rules and direction of future nurse migration. PMID- 9866292 TI - Nursing manpower development and strategic planning in Greece. AB - PURPOSE: A national study to register all working nursing personnel and nursing students; to estimate the nursing needs of the Greek population beyond the year 2000; to design a nursing resource master plan for the nursing needs of Greece. DESIGN: First, a questionnaire was distributed to register all nursing personnel. Second, required personnel were estimated according to the population of each of the country's regions. Third, a master plan for developing nursing personnel to the year 2010 was developed. RESULTS: The number of nursing personnel was found to be 35,715 which included 11,497 RNs and health visitors, 22,318 assistant nurses, 1,900 midwives. The number of students was 9,252. The majority of nursing personnel work in areas with the highest population. The need for nursing personnel was estimated to be 62,000: 36,300 RNs, 21,700 assistant nurses and 4,000 midwives. CONCLUSIONS: There is a serious shortage of RNs in the Greek health services resulting in a downgrading of nursing care quality. PMID- 9866293 TI - Estimating the market for nursing personnel in North Carolina. AB - PURPOSE: To facilitate nurse workforce planning in the United States at the state level, the North Carolina Center for Nursing conducted a statewide survey of nurse employers to describe the current market for nurses; identify the types of nursing personnel in short supply; estimate the effect of organizational changes on nursing demand; examine preferences for RNs with varying levels of education; and identify the specific skills or competencies desired by employers. DESIGN: Descriptive. METHODS: A geographically stratified random sample of 909 nurse employing organizations in North Carolina was surveyed in 1996 by telephone. A total of 667 interviews were completed for a response rate of 78%. Descriptive statistics and correlation analyses are reported. RESULTS: A nursing labor market was found characterized by uncertainty in hospitals, particularly those undergoing reorganization, but of definite growth outside of hospitals. Demand is increasing for unlicensed assistive personnel, RNs with baccalaureate degrees, some advanced practice RNs, and in hospitals, for RNs with master's degrees in management. Recruitment continues to be a major challenge to hospitals in specialty areas, particularly critical care and surgery. In the community sector, demand is strong for all types of nursing personnel. Tests for substitution of specific types of nursing personnel revealed no systematic substitution by employers. Critical thinking and management skills were competencies most valued by hospital employers while specific clinical competencies, including assessment skills and technical skills, received priority emphasis by community-based employers. CONCLUSIONS: Although change is occurring in the health system of North Carolina, there remains a strong demand for nursing personnel. The findings pose a challenge to produce a nursing workforce sufficient in numbers and education. PMID- 9866294 TI - When is the dyspnea worth it? Understanding functional performance in people with alpha-1 antitrypsin deficiency. AB - OBJECTIVE AND SIGNIFICANCE: To examine functional performance in people with emphysema because of alpha-1 antitrypsin (AAT) deficiency. A severe deficiency of AAT affects 1:3,500 to 1:1,670 Americans who can develop debilitating emphysema in the third to fifth decades of life. DESIGN: Exploratory. POPULATION: People with a severe deficiency of AAT. SAMPLE AND DEMOGRAPHICS: Thirty-three patients (21 men) with a mean age of 47 (SD = 7) years. YEARS: Data were collected 1993 1996. METHODS: Activities patients identified as important on the dyspnea subscale of the Chronic Respiratory Disease Questionnaire were categorized and interpreted within the context of an integrity framework (i.e., effectiveness--or connectedness-related). FINDINGS: Patients identified over 25 activities. Effectiveness activities, such as bathing, were mentioned most frequently (78%). Connectedness activities, such as playing with children, were mentioned less often (22%) but were ranked more important. Two patients reviewed and supported the content validity of the framework. CONCLUSIONS: Results offer insight into the activities patients with genetic emphysema choose to perform and the factors that influence the decision that "the dyspnea is worth it." IMPLICATIONS: Identifying the activities people with AAT deficiency choose to perform and understanding why these activities are meaningful can guide interventions to help patients maintain a sense of integrity. PMID- 9866295 TI - Hospital nurses' perceptions of the ethical climate of their work setting. AB - PURPOSE: To develop an instrument to measure how hospital nurses perceive the ethical climate of their work setting and to evaluate its psychometric properties. A way to measure ethical climate can help nurses understand the work setting influences on their practice and can be used to diagnose areas for organizational change as well as to evaluate effectiveness of organizational interventions. DESIGN: Descriptive. Instrument developed was the Hospital Ethical Climate Survey (HECS). Sampled were 360 RNs (48% response rate) employed in clinical practice at two acute-care hospitals in one U.S. midwestern city. The nurses completed a questionnaire in 1993-1994 consisting of items developed to assess their perceptions of ethical climate, along with the 43-item Integrity Audit, a social desirability scale, and a demographic inventory. METHODS: Construct validity was assessed by confirmatory factor analysis (CFA) using the LISREL 7 statistical program. RESULTS: The final CFA model consisted of 26 variables in five factors, which were organized according to the relationships of nurses with peers, patients, managers, hospital, and physicians. Internal consistency reliability, using Cronbach's alpha, for the final 26-item instrument was 0.91 (range 0.68 to 0.92) for the subscales. CONCLUSIONS: The Hospital Ethical Climate Survey has acceptable initial reliability and validity. PMID- 9866296 TI - Wellness self-care by healthy older adults. AB - PURPOSE: To examine self-care behaviors in healthy older adults. The number of older adults continues to increase. Nurses need a framework for understanding and supporting healthy behaviors in this population. DESIGN: Descriptive. METHOD: Data about self-care behaviors were elicited in 1997 about how healthy older adults stay well. Using a key-informant criterion and critical case approach, a small convenience sample of 28 older adults (ages 57-83) responded to a questionnaire based on a self-care wellness model. FINDINGS: Age was not related to differences in healthy behaviors. Many informants reported no digestive or sleep difficulties. All were active, regularly kept in touch with family and friends, were confident their environment was safe, obtained sufficient sleep and rest, and managed stress well. All but one attributed their quality of life and zest for living to remaining active, eating healthy food, exercising, pacing themselves, doing preferred activities, and reading the Bible, or "feeding the life of the mind." CONCLUSIONS: Participant statements of actions can provide inspiration for less-active seniors and a beginning framework for nurses for understanding and supporting wellness self-care behaviors in older adults. PMID- 9866297 TI - Factors associated with domestic violence in low-income Lebanese families. AB - PURPOSE: To analyze the cultural context of domestic violence in low-income Moslem and Christian-Armenian families living in Lebanon. Analysis also included an identification of family stressors, conflict management strategies, and Moslem and Christian-Armenian cultural differences. The study was part of a larger project designed to identify patterns of wife and child abuse in low-income Middle Eastern families living in Lebanon and Egypt. DESIGN: Qualitative content analysis of descriptive narratives by 60 low-income women who self-reported spouse abuse in two urban Lebanese clinics during a 2-month period in 1992. METHODS: Narratives describing exemplary incidents were obtained during a semi structured interview and recorded in the participant's native language then translated to English for coding and content analysis. FINDINGS: Contextual factors for violence included unmet gender role expectations, conflict with husband's relatives, and alcohol abuse. Family stressors were: emotional, financial, and work. Women used three types of conflict management: negotiation, taking initiative, and passive resignation. CONCLUSION: From a cultural perspective, the analysis revealed both strengths and vulnerabilities of Lebanese women who experienced domestic violence. The study raised several questions, including whether it is appropriate to apply Western-generated domestic violence theories to a Middle Eastern population. Culturally-specific nursing interventions should be directed toward bolstering strong family and social resources to cope with family stressors and to modify patterns of maladaptive communication. PMID- 9866298 TI - Nursing research in Finland from 1958 to 1995. AB - PURPOSE: To review the methods and content of nursing research carried out in Finland between 1958 and 1996. University-level nursing education and research started in Finland in 1979. In 1997, there were seven universities with nursing programs and, in all of them, active international collaboration. SCOPE: First emperical and nonemperical studies were identified, then the research topics and research methods were analyzed. Some historically interesting evaluations were also documented. SOURCES: Articles published in the Yearbook of Nursing (1958 1988, n = 228), in the Finnish Journal of Nursing Science (1989-1995, n = 116), and all Finnish licentiate and doctoral theses (n = 104). METHOD: Content analysis. FINDINGS: The number of research articles and theses is increasing. Nursing practice has been the most common research topic, and especially primary health care and surgical and medical care. Questionnaires and interviews are the methods used most often, observational methods are rare. Most research is discriptive. CONCLUSIONS: In the future, research is recommended in nursing care of children and adolescents as well as evaluative research. PMID- 9866299 TI - Mapping nursing diagnosis nomenclatures for coordinated care. AB - PURPOSE: To map the problem or diagnosis terms from three nomenclatures, term to term, to determine commonalities and differences; and to determine whether it is possible to develop a single vocabulary that contains the best features of all. When different nomenclatures are used in different settings, continuity of care is hampered by the need to re-state problems and interventions. DESIGN: The sample for this descriptive analysis was 396 terms from three nursing diagnosis and problem nomenclatures recognized by the American Nurses Association: the North American Nursing Diagnosis Association (NANDA) Approved List, the Home Health Care Classification (HHCC), and the Omaha System. METHOD: Terms from each of the three nomenclatures were mapped to terms in each of the others. Consensus methods were used to resolve differences in mapping decisions. Terms were characterized as "Same," "Similar," "Broader," "Narrower," and "No Match." Validation of consistency and accuracy was done by reverse mapping, use of syllogisms, use of taxonomic groupings, and expert review. RESULTS: Of 396 terms, 21 concepts accounting for 63 terms were found to be the same or similar in all three nomenclatures; 91 terms were unique to the nomenclature in which they were found ("No Match"). The remaining 242 terms had a narrower or broader relationship to at least one term in another nomenclature. In all three nomenclatures, inconsistencies existed in level of abstractness of the diagnosis or problem terms, and in definition and placement of terms within their own taxonomic structure. CONCLUSION: Because of differences in structure and incompatible taxonomic arrangements, a master list of "preferred terms" taken from the three nomenclatures is not feasible. However, the mappings are useful for determining commonalities and the unique contributions of each nomenclature, which can facilitate the development of a uniform language for nursing diagnoses. The mapping can also form the basis for automatic translation of computer-stored nursing diagnoses from one setting to another when different nomenclatures are used. PMID- 9866300 TI - Addressing urinary incontinence with educational continence-care competencies. AB - This article is a report of a task force formed from a discussion group hosted by the American Nurses Association on the AHCPR Guidelines on Urinary Incontinence. The need to standardize content related to bladder incontinence to be used in curriculums was identified as an initial step in implementing the guidelines. A task force was formed to develop educational competencies to be used by schools in identifying content that should be addressed at various levels of preparation. Although special training is needed when continence care is a significant part of a nurse's practice, it is expected that by using the educational competencies, schools will produce graduates at the basic level able to provide beginning continence care and graduate nurses able to address advanced incontinence problems. PMID- 9866301 TI - Historic and future health promotion contexts for nursing. AB - PURPOSE: Health promotion in the United States has been narrowly defined focusing primarily on individual behaviors, risk factors, and lifestyle. This article traces the historic and conceptual roots of health promotion care in the United States and critically examines the direction nurses have taken in health promotion. SCOPE: Health promotion care in Western ideology emphasizes the value of individuals and the importance of personal responsibility for success or failure. CONCLUSIONS: Health promotion nursing interventions, based on the tenets of individual choice and knowledge, restrict the range of interventions. The complex nature of health promotion requires that nurses consider the economic, sociopolitical, and cultural context in which health care takes place. Recommendations are made for an expanded nursing role in health promotion, one that is consistent with nursing's underlying contextual focus and concern with environments. PMID- 9866302 TI - Case study of institution-building by nurse Bertha Wright and colleagues. AB - PURPOSE: To illuminate the history of nurses' participation in institution building in the early 1900s, using the Baby Hospital in Oakland, California (now called Children's Hospital Oakland) as a case. DESIGN: Historical research using a framework of women's history theory focused on the women who founded Oakland's Baby Hospital, 1910-1930. METHOD: Data collection included studying original sources, archival material, and interviews. Data evaluation included external criticism for authenticity and genuineness, and internal criticism for accuracy and bias. FINDINGS: In 1912, nurse Bertha Wright and social worker Mabel Weed, with a circle of community women, including Jessica Peixotto and Jean Howard McDuffie, established the Baby Hospital. Their activism included home visits, education, research, publications, political lobbying, and state policy and program development. At the center of this activity, was the lifelong commitment of Wright and Weed, who created new definitions of family. With social changes in the late 1920s, the male Board of Directors seized control of the hospital, and relegated women to auxiliary roles. CONCLUSIONS: Health and social issues of the United States at the turn of the century are still problematic as a new century approaches and pertain to maternal-child health, foster care, and immigration. Feminists, including nurses, have played a central role in creating solutions. Knowledge of some of their incredible effort has been lost, particularly nursing's history in the western United States. PMID- 9866303 TI - [The aspects of erosive-ulcerative changes of the gastroduodenal tract mucous membrane in atherosclerosis of the aorta and major arteries]. AB - Complex clinico-instrumental examination and surgical treatment of 172 patients with erosive-ulcerative changes of the gastroduodenal zone mucosa and atherosclerotic affection of aorta and main arteries was conducted. Functional peculiarities of erosive-ulcerative changes of the gastroduodenal zone mucosa in atherosclerosis of aorta and main arteries were revealed. PMID- 9866304 TI - [Aspects of excision of the perforative duodenal ulcer complicated by penetration and stenosis]. AB - The results of treatment of 259 patients with perforative duodenal ulcer, complicated by penetration and stenosis, were analysed. The original methods of ulcer excision are proposed. Excellent and good late follow-up result was noted in 159 (84.6%) patients, fair--in 15 (8.2%), bad (ulcer recurrence)--in 8 (4.4%). PMID- 9866305 TI - [The prognosis of the duodenal ulcer course]. AB - The method is proposed, permitting to prognosticate the ulcer disease course, to specify the volume of planned therapy and operative intervention, basing on objective data, obtained while biochemical and immunological investigations of gastroduodenal parietal wall mucus, the fucoglycoproteids and secretory immunoglobulin A contents, histological diagnostic of dissemination with Helicobacter pylori. PMID- 9866306 TI - [Pharmacotherapeutic correction of gastric motility and emptying after gastric resection and vagotomy]. AB - Motor-evacuational function of stomach, duodenum and its draining loop after gastric resection and vagotomy in 82 patients is studied up. Two variants of medicinal correction of the stomach and efferent intestinal loop motor disorders are elaborated. PMID- 9866307 TI - [Clinical-statistical prognosis of gastroduodenal erosion and ulcer incidence after surgical treatment of inflammatory disease of hepato-biliary tract]. AB - An acute postoperative erosive-ulcerative bleeding gastroduodenal mucosal affections were diagnosed in 40 patients of 410, operated on for inflammatory diseases of the biliferous tracts. The method of individual prognosis of the disease occurrence was proposed. PMID- 9866308 TI - [The effectiveness of a comprehensive treatment of ulcers]. AB - Complex treatment of 693 patients with gastroduodenal ulcer disease was conducted before the operation, in the early, late and remote periods after the surgical intervention. The comparative results of treatment of 373 patients in terms up to ten years were analysed, depending on the matter of the complex treatment and sanatorium sanitation conduction done and in 288-without one. Positive result was noted accordingly in 361 (96.8%) and 257 (89.2%) patients, bad one--in 12 (3.2%) and 31 (10.8%). PMID- 9866309 TI - [Translocated gastroduodenal anastomosis in surgical treatment of penetrating duodenal ulcer]. AB - The method of operative treatment of complicated duodenal ulcer disease consisting gastroduodenostomy, translocated by 3-4 cm medially and lower from the ulcer crater and strengthened by a spur on a small curvature side. PMID- 9866310 TI - [Re-operations after formerly performed suturing of the "silent" perforative duodenal ulcer]. AB - After the "silent" perforative duodenal ulcer closure the gap till the complications would occur, which need reoperation, by two times more than such period of time in patients with typical ulcer anamnesis, preceding the perforation. The reoperation causes are the recurrence (in 50.6% of observations), newly occurred ulcer (in 26.9%) or nonhealing of already existing (in 22.5%) duodenal ulcer. The reoperation method of choice is gastric resection according to Billroth-I in combination with truncal vagotomy in case of hypersecretory syndrome. PMID- 9866311 TI - [Postop complications of perforative ulcer surgery]. AB - Of 342 patients operated on for gastroduodenal perforative ulcer the complications have occurred in 63 (18.4%) and a progressive peritonitis in particular--in 45 (71.4%). In 4 (2.8%) of patients a peritonitis progress was noted after truncal vagotomy and ulcer excision conduction without the infection toxical shock signs. PMID- 9866312 TI - [Modern approaches to surgical treatment of peptic ulcer of gastroenteroanastomosis]. AB - The results of operative intervention in 157 patients with recurrent peptic ulcer of gastroenteroanastomosis are studied up. The advantages of vagotomy with the ulcer substrate excision or the gastric remnant reresection are noted. PMID- 9866313 TI - [The use of vertical gastroplasty in surgical treatment of obesity]. AB - Vertical gastroplasty was applied in surgical treatment of obesity in two patients. There were no complications. During the first year after the operation the patients body mass have reduced by 35-41 kg (25-29% of initial). PMID- 9866314 TI - [Surgical strategies in combined cicatricial stenosis of esophagus and stomach after chemical burn]. AB - The results of treatment of 78 patients with the cicatricial esophageal and gastric exit stenosis are adduced. The tactics chosen have foreseen the feeding fistula and gastroenteroanastomosis conduction with subsequent substernal colonic esophagoplasty. The characteristics of different feeding fistula, the advantages and faults of every of them are adduced. PMID- 9866315 TI - [Methods of intestinal drainage for an acute mechanical ileus]. AB - There were examined 362 patients with acute small intestine ileus (SII). It was established, that retrograde intubation according to Zhitnyuk is necessary to apply while treating low SII, especially in patients of elderly and senile age with concomitant cardiovascular and respiratory systems diseases. In patients with an adhesive SII the application of nasogastrointestinal intubation in combination with respiratory therapy is indicated. The jejunostomy with one-stage intestinal intubation via enterotomic hole application is mandatory for high SII, especially if an early enteral feeding is necessary. One-stage intestinal intubation via enterotomic hole as the independent method may be applied for intestinal preparation before the one-stage radical operation conduction. PMID- 9866316 TI - [Dysbiotic changes of colonic microflora in patients with Crohn's disease]. AB - In patients with Crohn's disease the dysbiotic changes in colonic lumen was revealed, what was testified by reducing the quantity of saprophytic and increasing the quantity of the conditionally-pathogenic microorganisms. The application of "Salofalk" preparation do not influence the colonic microflora composition. PMID- 9866317 TI - [Acute biliary pancreatitis. Diagnosis and course of treatment]. AB - The results of surgical treatment of 320 patients with an acute biliary pancreatitis are analysed. The therapeutic-tactical algorythm, using miniinvasive endoscopical interventions, is elaborated. Mortality for pancreatitis in systemic toxical phase have constituted 10.1%, and in septical one--33.3%. PMID- 9866318 TI - [Splenic blood circulation in liver cirrhosis with portal hypertension syndrome]. AB - The result of the hemodynamics in spleen studying in 90 patients with liver cirrhosis and a portal hypertension syndrome is adduced. The significance of hyperdynamic state of blood circulation in spleen, influencing the portal hypertension occurrence and the secondary hypersplenism formation, was established. The conversion of liver cirrhosis to decompensation stage is caused by the change of splenic circulation from hyperdynamic state to congestive one. PMID- 9866319 TI - [Results of application of endolymphatic polychemotherapy and local supra-high frequency hyperthermia in comprehensive treatment of rectal cancer]. AB - The treatment efficacy of a widespread cancer of recti using neoadjuvant endolymphatic polychemotherapy and local suprahigh frequency hyperthermy in a radically operated patients was studied up. In patients without regional metastases the five-year survival index have increased from 63 to 81%, and the recurrencies occurrence frequency have lowered from 20 to 10%. The method proposedhas small efficacy in patients with metastases in regional lymph nodes. PMID- 9866320 TI - [The conduction of combined operations for pulmonary cancer]. AB - The supraexpanded (combined) operation was conducted in 36 patients for pulmonary cancer. The conduction of such operations is indicated in spite of their significant postoperative complications occurrence and high postoperative mortality (22.2%) because it permits to improve a grade II pulmonary cancer result of treatment. PMID- 9866321 TI - [Results of treatment of oral cavity mucosa cancer according to the data of the regional oncological clinic]. AB - The results of treatment of 139 patients with mucosal cancer of oral cavity are summarized. The most effective method of treatment is combined one, including the operative intervention conduction. In 18 patients intraoperative cryoimpact on tumor was conducted, permitting to reduce the recidives frequency and to increase their life span. PMID- 9866322 TI - [Aspects of etiology, pathogenesis and clinical course of the megaureter]. AB - Complex examination was conducted in 408 patients with megaureter. Five stages of the disease are delineated. Greatest importance in the megaureter pathogenesis play the peristalsis disorder and the ureter's ectasia, the rise of intraluminal pressure, which are the compensational factors, directed on the urodynamics and renal function support. PMID- 9866323 TI - [Investigation of the brain reactiveness depending on the type of human intrinsically present electroencephalography]. AB - While adopting the lighting flashes 9 Hz in frequency, and while the absence of the adopting the rythm reaction during the photostimulation conduction in majority of the patients examined the alpha-oscillations amplitude according to background electroencephalography constitutes 25-50 mcV; while adopting of the lighting flashes rythm with 6 and 12 Hz frequency--50-75mcV. PMID- 9866325 TI - [A clinical significance of of methods of the colonic motor function investigation]. PMID- 9866324 TI - [Postvagotomy adaptation syndrome]. AB - It was established in experiment, that the changes of the natural resistance of organism indexes and of the peritoneal cavity cytology has compensatory adaptational character while the denervation-adaptational syndrome occurrence and progress, which may be assessed as eustress. Vagotomy and operative trauma cause qualitatively different reactions of an organism. PMID- 9866326 TI - [Radical operation in patients with a recurrent suturing of the perforative duodenal ulcer]. PMID- 9866327 TI - [Two-stage method of surgical flaps as the choice method in the surgery of replacing defects of limb tissues in unusual clinical situations]. PMID- 9866328 TI - [The incidence of the perforative bleeding ulcer of the duodenum in a combination with pancreatic necrosis]. PMID- 9866329 TI - [Cholelithiasis as a cause of acute intestinal obstruction]. PMID- 9866330 TI - [The choice of surgical intervention method for gastroduodenal perforative ulcer and its connection with progressing peritonitis in the early postop period]. PMID- 9866331 TI - [Penetration of gastroenteroanastomosis peptic ulcer in anterior abdominal wall stimulating a tumor]. PMID- 9866332 TI - [Durable asymptomatic stay of foreign body in esophageal tract]. PMID- 9866333 TI - [Cholelithiasis and small intestine obstruction]. PMID- 9866334 TI - [The treatment of nephrolithiasis of doubled kidney and of the upper urinary tract]. PMID- 9866335 TI - [Selective intraarterial application of Thienam and Vasoprostan in the treatment of purulent foot diseases in patients with diabetes mellitus]. PMID- 9866336 TI - [Surgical analysis for metastatic lung tumor from renal cell carcinoma]. AB - Metastatic lung tumor from renal cell carcinoma were studied in 29 cases. Eighteen patients were treated surgically, 11 were treated non-surgically. The overall 5-year survival rate with the patients of pulmonary resection was 53.5%, and that with those of conservative therapy was 0%, and this difference is statistically significant (p < 0.05). There was no significant difference in any characteristics such as sex, age, stage, grade, disease free interval, metastatic pattern and combination with or without interferon therapy. There was no significant difference in surgically treated patients with pulmonary metastasis in terms of any factors such as age, sex, stage, grade, disease free interval, pulmonary metastasis pattern, metastatic number, surgical procedure, combination with or without interferon therapy statistically. Analysis for the surgically treated patients with pulmonary metastasis from renal cell carcinoma shows no significant difference in prognosis with any characteristics. This result shows efficacy of surgery even if for the patients with synchronous bilateral multiple pulmonary metastasis from renal cell carcinoma. PMID- 9866337 TI - [Surgical treatment of an aortic arch aneurysm: reports of 2 cases with lesion of the arch vessels]. AB - A 48-year old woman underwent surgery for an aortic arch aneurysm with stenosis or dilatation of three arch vessels caused by aortitis syndrome. Total arch replacement and reconstruction of three arch vessels were performed with hypothermic selective cerebral perfusion (SCP). To avoid atheroembolism and malperfusion to the brain, the rt. common carotid artery was perfused via the rt. subclavian artery through the dacron vascular graft and the lt. subclavian artery was cannulated and perfused distally to stenosis. A 65-year-old man who had an atherosclerotic aortic arch aneurysm with severe stenosis of the brachiocephalic artery underwent operation. In the operation, extracorporeal circulation was instituted with the arterial return through the lt. subclavian artery. Same as case 1, total arch replacement and reconstruction of three arch vessels were performed under hypthermic SCP. In this case, the left common carotid artery was transected and cannulated directly into the vessel, instead of cannulation through the aortic lumen because of atheromatous plaques in the orifice of the left common carotid artery. The patients recovered uneventfully and doing well now. PMID- 9866338 TI - [Simultaneous cholecystectomy and CABG for acute cholecystitis and post infarction angina]. AB - A 71-year-old man was admitted to our hospital with massive GI bleeding and followed by acute myocardial infarction. Severe three vessel coronary artery disease with poor left ventricular performance (EF: 40%) was demonstrated by coronary angiography. He was referred for CABG. He had also cholecystitis, which needed surgical treatment simultaneously. Combined CABG and cholecystectomy were performed without using gastroepiploic artery as a bypass graft, and through separate median sternotomy and through right upper transrectal laparotomy. Postoperative course was uneventful. This combined procedure is beneficial for the patients with acute cholecystitis and unstable angina. PMID- 9866339 TI - [The long-term anterior chest median wound keloid after the cardiac surgery]. AB - We discussed on the long-term anterior chest median wound keloid after the cardiac surgery by using a questionnaire. The risk of the anterior chest median wound keloid was high significantly in the cases using the bilateral internal thoracic artery compared with the cases using no internal thoracic artery. Participation of the decreased blood supply was highly suggested as the cause of this keloid formation. PMID- 9866340 TI - [Dorsal sympathectomy for palmar hyperhidrosis by the thin thoracoscope]. AB - The purpose of this study was to examine the efficiency to use by the thin thoracoscope for the palmar hyperhidrosis. General anesthesia with double lumen endotracheal intubation was used in all cases. A 2 mm incision made in the anterior axillary line in the third intercostal space. Pneumothorax was obtained by insufflation 1.5-2 L of CO2. A 2 mm endoscopic trocar was inserted through this incision, and a 2 mm, 0-degree scope (Autosuture, USA) was introduced. A second 2 mm trocar was inserted in the middle axillary line in the fourth intercostal space, through which a straight endoscopic seizer was introduced. The chain was dissected by electrocutting the white and gray rami and was incised over the second and third ribs. To avoid puemothorax, 8 F thoracic catheter was introduced through a guide wire which was inserted through a 2 mm trocar. The lung was expanded, and then the thoracic catheter was removed. We performed six sympathectomies on three female patients. All patients was satisfied of their results. This technique used by thin thoracoscope was cosmetic and a skin incision did not need to close too small. PMID- 9866341 TI - [Supervised cardiac rehabilitation for the patients with the disturbance of the motor function after cardiac surgery: the significance of physical therapists' participation]. AB - There are sporadic instances of patients with the motor function disturbance of non-cardiac origin after cardiac surgery, and these patients may need prolonged post-operative cardiac rehabilitation. We established our cardiac rehabilitation program for post-operative patients and a total of 124 patients underwent the post-operative cardiac rehabilitation program (male 73, female 51, average age 60). Among them, 12 patients (9.7%) received the physical therapy for the disturbance of motor function post-operatively. These 12 patients were retrospectively studied. Physical therapies performed were the exercise therapy to improve the range of motion to prevent contracture in 3 patients with peroneal nerve palsy and drop foot, the exercise therapy for pre-operative or post operative hemiplegia in 6 patients, the instruction of exercise for lumbago in 1 patient with spinal cord disease, respiratory physical therapy in 1 patient, and myotherapy for arthritis in 1 patient. Treatment with physical therapy was very useful in rehabilitating these patients. Active participation of physical therapists in cardiac rehabilitation for patients with the disturbance of motor function after cardiac surgery is possible. It is expected that their participation may improve the quality of life in this subset of patients. PMID- 9866342 TI - [Evaluation of the effect of ischemic preconditioning using near-infrared spectroscopy]. AB - Ischemic preconditioning (IP) protects the myocardium from subsequent sustained ischemic insults. Temporary occlusion of the coronary artery is indispensable for anastomosing the graft vessel during coronary artery bypass operation without cardiopulmonary bypass. In the canine model of ischemia and reperfusion, we measured myocardial tissue oxygen saturation (SO2) continuously using near infrared spectroscopy to determine the effect of IP on myocardial oxygen metabolism. Nine dogs underwent occlusion of the left descending coronary artery for three 5-minute periods, followed by three 5-minute periods of reperfusion. The dogs were then subjected to a 20-minutes periods of sustained coronary artery occlusion, followed by prolonged reperfusion. The myocardial SO2 was 82 +/- 2% at the baseline before coronary occlusion and was decreased to 74 +/- 2%, 76 +/- 2%, 77 +/- 3%, 77 +/- 3% at the first, second, third and sustained coronary occlusion, respectively. The increase in the minimum myocardial SO2 value at the second and third coronary occlusion suggested the effect of IP. Near-infrared spectroscopy is a useful method of continuously monitoring myocardial oxygenation and of evaluating the effect of IP during off-pump heart surgery. PMID- 9866343 TI - [Influence of left ventricular wall thickening on myocardial injury in aortic valve replacement]. AB - We retrospectively examined the influence of left ventricular wall thickening on myocardial injury in aortic valve replacement. Thirty-five patients who underwent aortic valve replacement in our hospital between January 1995 and July 1997 were studied. We divided these patients into 3 groups: group N (left ventricular thickness (LVT) < or = 10 mm), group H (10 < LVT < or = 15 mm), and group S (LVT > 15 mm). All patients of group S had aortic stenosis. CK and CK-MB were significantly higher and the cardiac index was significantly lower in group S than in the other groups. Intra-aortic balloon pumping was required for 50% of patients in group S, and a total dose of catecholamine above 10 micrograms/kg/min was also required for 50% of patients in this group. In the other groups, these percentages were significantly lower. Myocardial protection and postoperative care were difficult for patients with severe left ventricular hypertrophy with LVT over 15 mm in this study. Left ventricular wall thickness was a more important factor in determining difficulty with intraoperative myocardial protection than was left ventricular mass. PMID- 9866344 TI - [Long-term survival in surgical cases of non-small cell lung cancer with mediastinal nodal involvement]. AB - We studied long-result of the patients with mediastinal lymph node metastasis (N2) in lung cancer. From 1981 to 1991 in National Sanatorium Nishi-Gunma Hospital, we have performed a lobectomy or pneumonectomy with mediastinal lymph node dissection for 306 patients with non-small cell lung cancer. Six operative deaths were excluded (operative death rate: 2.0%). Of these, 53 patients (17%) were diagnosed as adenocarcinoma or squamous cell carcinoma with N2 disease pathologically. The patient consisted of 31 men and 22 woman whose ages ranged from 18 to 73 years, with a mean age of 58.1 years. The results were: Mean survival period was 39 month. Five-and 10-year survival rates were 18.9% and 12.6%, respectively. By motive of detection, the group of following other disease had a better prognosis than the group of having symptom. By pT-factor, T1 had a better prognosis than T3. By the number of nodal involvement level, 1 level had a better prognosis to compared to 3 or more than 4 levels. There were 10 patients of 5-year or over survivors after surgery, none of whom had primary site in left upper lobe. Analysis with a Cox's proportional hazard model demonstrated that the number of nodal involvement level was a useful factor in prognosis. PMID- 9866345 TI - [The investigation of surgical procedures for post infarcted ventricular septal defects: a comparison between Daggett and David method]. AB - Post infarcted ventricular septal perforations (VSP) has diverse clinical and pathological manifestations and the surgical results in severe cases have not yet been satisfactory. In the past we had performed the procedure of Daggett. However since 1992 we have also introduced the procedure of infarction exclusion technique proposed by David which involves plastering the ventricular cavity in a large patch without infarcted myotomy. We reported 5 cases treated with David method, 5 cases with Daggett method, and their results were compared. There were four mortality cases in Daggett method and three in David method. No significant difference was observed regarding the surgical mortality. However, there were different post-operative features between the two groups. In Daggett method patients had prolonged heart failure and LOS. In David method patients, had no severe LOS, but residual shunt were recognized in two cases. We think that the emergence of residual shunt in David method was probably due to suturing without prior removal of the damaged fragile infarcted myocardium. In Daggett method there were death cases caused by LOS and postoperative heart failure but without residual shunts. Despite its recent growing popularity for treating VSP, our experience suggests that residual shunt is an important issue remains to be solved in David method. PMID- 9866346 TI - [Evaluation of an ultrasonic cutting and coagulating system (harmonic scalpel) for performing a segmental and wedge resection of the lung]. AB - In order to prevent the local recurrence of malignant tumors, it is important for surgeon to maintain a sufficient margin between the tumor and the edge at resection. For this reason we do not use an auto-suturing device, but instead use the ultrasonic cutting and coagulating system (HARMONIC SCALPEL, ETHICON ENDO SURGERY Cincinnati, Ohio) whenever we perform either a segmental resection or a wedge resection of the lung. The subjects investigated consisted of 24 cases of lung tumors (15 metastatic tumors, 5 cases with primary lung cancer, 3 inflammatory tumors; and one benign tumor). The type of operation included 10 segmental resections and 14 wedge resections, with 21 open thoracotomies and 3 instances of thoracoscopic surgery, while 15 were single resections and 9 were multiple resections. Little bleeding was seen at the resection of the parenchyma and the vessels of the lung. However prolonged air leakage was observed in some cases that needed pleurodesis. The mean duration time of the surgery was 266 minutes, and the mean blood loss was 173 ml. The operative duration was a little longer than normal, because many cases were not first thoracotomies and some cases had multiple tumors. Nevertheless the amount of blood loss was slight. The longest post-operative period was two years and six months, no local recurrence has yet been seen in any of malignant cases. We consider this system to be very effective for performing a resection of the lung parenchyma because of the reduced blood loss and the apparent increased prevention of recurrence. PMID- 9866348 TI - [An emergent surgical treatment of recurrent Stanford type A thrombosed aortic dissection]. AB - A 71-year-old male was admitted to our hospital with a chief complaint of back pain. Chest CT scan showed so-called Stanford type A thrombosed aortic dissection. Aortography revealed no ulcer like projection. At first, conservative treatment was carried out. But about two hours later, he had another back pain. Repeat chest CT scan demonstrated an enlarged dissecting lumen, necessitating emergent graft replacement. Postoperative course was uneventful and he was discharged on the 33rd postoperative days. PMID- 9866347 TI - [A case report of infective endocarditis with total rupture of the posterior papillary muscle after aortic valve replacement]. AB - We reported a case with severe mitral regurgitation caused by total rupture of the posterior papillary muscle two days after aortic valve replacement. A 62-year old man was transferred to our hospital with high fever and dyspnea with severe aortic regurgitation caused by infective endocarditis. The left heart failure occurred suddenly two days after the initial operation. Echocardiogram revealed massive mitral regurgitation and rupture of the posterior papillary muscle. He underwent emergent mitral valve replacement. Histological examination of the papillary muscle showed typical ischemic necrosis without inflammation. The postoperative course was uneventful. We suggested the papillary muscle rupture in this case may be due to coronary artery emboli occurred in association with infective endocarditis. PMID- 9866349 TI - [Relative valve area stenosis after replacement of a Starr-Edwards mitral valve (model 6520) for corrected TGA (S, L, L): report of a case]. AB - The patient was a 38-year-old man who had undergone mitral valve replacement with a Starr-Edwards disc valve (model 6520, 3 M, phi 30 mm, functional valve area: 2.85 cm2) at the age of 15 years for TR combined with corrected TGA. He developed dyspnea on exertion and palpitations at the age of 30 years, and paroxysmal supraventricular tachycardia appeared at the age of 35 years. Cardiac catheterization revealed that the PCWP was 30 mmHg and the TVA was 1.1 cm2, and a diagnosis of stenosis of the prosthetic valve was made. Thus, replacement of the Starr-Edwards disc valve using a BICARBON double leaflet valve (phi 27 mm, functional valve area: 2.85 cm2) was performed. The operative findings of the Starr-Edwards disc valve revealed a normal appearance and no pannus growth. Postoperative echocardiography and cardiac catheterization found that the TVA was 3.7 cm2 and the PCWP was 13 mmHg. The actual valve area of the disc valve sewn on the ventrical, which was anatomically RV construction might have been smaller than the ideal area; however, the double leaflet valve proved efficient in our patient. This case report serves to demonstrate that because of the postoperative difference in measurement between the actual valve area and the prostheticin ball or disc valve, patients undergoing mitral valve replacement must be followed up carefully. PMID- 9866350 TI - [A surgically treated case of saccular true aneurysms in the ascending aorta]. AB - A surgically treated case of saccular true aneurysms in the ascending aorta caused by cystic medial necrosis is reported. A 59-year-old woman, who had no findings of Marfan syndrome, was admitted to our hospital because of chest discomfort and dyspnea on exertion. Echocardiography showed that massive pericardial effusion and the dilated ascending aorta without dissection. No aortic valve insufficiency nor dilatation of the Valsalva's sinuses were found out. Chest CT scan and aortography also revealed that focal dilatation of the ascending aorta without the intimal flap and the false lumen. Because of the presence of cardiac tamponade, we performed an emergency operation without definite diagnosis of the ascending aortic lesion. Intraoperatively, 2 saccular aneurysms, which were 55 x 45 x 20 mm and 25 x 15 x 15 mm in size, were found out in the ascending aorta. The ascending aorta was replaced with a woven Dacron graft successfully. These aneurysms were histologically diagnosed as true aneurysms caused by cystic medial necrosis. Although saccular true aneurysms of the ascending aorta are rarely observed, they should be considered as one of ascending aortic diseases that cause cardiac tamponade potentially. PMID- 9866351 TI - [Surgical removal of right atrial myxoma with severe hypoxemia caused by right to left shunt through persistent foramen ovale]. AB - A 41-year-old woman was referred to our hospital due to dyspnea and cyanosis. Echocardiography revealed a right atrial tumor with right to left shunt through PFO. There was no evidence of pulmonary embolism from the tumor. The tumor was removed under cardiopulmonary bypass and PFO was closed. The patient is doing well now. PMID- 9866352 TI - [A case of video-assisted thoracic surgery for congenital esophago-bronchial fistula]. AB - A 42-year-old male was admitted to our hospital because barium esophagograpm showed an esophago-bronchial fistula with an esophageal deverticulum. He has had frequent episodes of cough at drinking water since childhood. A chest CT scan showed mild inflammatory change and bronchiectasis in the right S6. Division of the fistula by video-assisted thoracoscopic surgery (VATS) was performed. There was no evidence of inflammation and adherent lymph nodes around the fistula. This case was diagnosed as a congenital esophago-bronchial fistula by operative findings and clinical course. The fistula was dissected easily and divided by an auto-suturing instrument. The affected lung could be preserved. The patient was discharged on the 10th postoperative day. VATS is an effective treatment for the patients of congenital esophago-bronchial fistula (Braimbrige type I and II), if they are not accompanied with pulmonary abscess or pleural empyema. PMID- 9866353 TI - [Two cases of metastasizing benign leiomyomatosis from myoma uteri]. AB - Case 1. A 50-year-old woman was referred to our hospital because of multiple bilateral small round lesion on chest radiography. She had undergone total hysterectomy for myoma uteri at the age of 33. She underwent thoracoscopic tumor excision at left lung. The lesion was proved benign. Right side lesions were laterly excised using thoracotomy. Case 2. A 49-year-old woman was referred to us because of two ovoid lesion at left lung area of chest radiography. She also had undergone total hysterectomy for myoma uteri at the age of 37. She underwent tumor extirpation using thoracotomy. All samples of two patients revealed, pathologically, lesions were consisted of benign spindle-like calls similar to those of myoma uteri. Therefore, we consider these lesions were pulmonary metastasis of myoma uteri. Myoma uteri has certain potential of metastasizing to the lung, in spite of benign disease. PMID- 9866354 TI - [A case of intrathoracic lipoma arising from the chest wall resected by video assisted thoracoscopic surgery]. AB - We present a case of lipoma arising from the chest wall spreading into the thoracic cavity. Although asymptomatic, a 65-year-old female was pointed out an abnormal shadow on the chest X-ray film taken at the mass survey. The tumor, measuring 3.4 x 3.0 x 2.0 cm in diameter, was surgically removed under thoracoscopic visualization through a small thoracotomy incision of lt. 1st intercostal space, and the diagnosis of lipoma was confirmed postoperatively by histopathologic examination. PMID- 9866355 TI - Re: "Perceptions" of questionable value. PMID- 9866356 TI - Patterns of death in world leaders. AB - This study presents an analysis of the mortality patterns of people who become world leaders. Using information in the public domain, we identified 261 world leaders who died between 1965 and 1996. Of these, 118 died while in office, 44% violently, often by assassination. Of the 143 leaders who died after leaving office, 11% died violently. The violent deaths occurred worldwide but most frequently in the Middle East/South Asia and sub-Saharan Africa regions. The most frequent natural causes of death among world leaders were heart disease, cancer, and stroke. Mortality patterns reveal that the longevity of those leaders who died of natural causes could have been predicted by U.S. life tables. This study suggests that world leaders are neither biologically "tougher" nor more vulnerable to disease than others; however, their odds of dying violently while in office are high. PMID- 9866357 TI - Leishmaniasis in the United States military. AB - Leishmaniasis is a recurrent health problem for the U.S. and other militaries. Health care workers may be unfamiliar with the risk factors, transmission, clinical features, diagnosis, and treatment of this disease. A team of highly trained specialists is required to properly manage service members with leishmaniasis. Such care is available only in a few medical centers. Although there are no prophylactic drugs to prevent this disease, control of insect populations and use of personal protection measures can minimize arthropod related casualties. The impact of leishmaniasis on military operations and research initiatives to better prevent, diagnose, and treat infection are discussed. PMID- 9866358 TI - Ambulatory urodynamics of female soldiers. AB - The purpose of this study was to assess the accuracy of ambulatory urodynamic monitoring compared with conventional urodynamic studies for the detection of exercise-induced urinary incontinence in the female soldier. Fifty active duty female soldiers with exercise-induced urinary incontinence and 10 asymptomatic control soldiers underwent conventional multichannel cystometry and then ambulatory monitoring during work or exercise. Ambulatory monitoring detected a greater number of abnormalities than conventional multichannel urodynamic studies in exercise-induced urinary incontinence. This greater sensitivity is valuable in formulating more effective treatment. Behavioral interventions were effective in treating exercise-induced urinary incontinence in this population. Test results normalized after behavioral intervention. It is neither cost-effective nor efficacious to require sophisticated urodynamic testing before instituting behavioral interventions. PMID- 9866359 TI - Satisfaction of active duty U.S. service members with military dental care. AB - This study explores the satisfaction of active duty service members with military dental care. The data were collected from April 1994 to January 1995. A prestratified, random sample of 15,915 service members was drawn. Of 12,050 respondents (81% response rate), 11,200 had an annual dental visit and answered 22 questions on satisfaction with military dental care. We calculated simple descriptive statistics and derived a composite overall satisfaction score using factor analysis. We then converted composite scores into a satisfaction index, a continuous variable with a mean of 100 and a standard deviation of 10. The satisfaction index was regressed on respondent demographics to determine which factors influence service members' overall satisfaction with military dental care. Results show that satisfaction with military dental care is high and consistent across respondent demographics. However, because this study was conducted before the recent, sizable drawdown of military dental personnel, these findings may not describe the current state of satisfaction with military dental care. PMID- 9866360 TI - Determinants of dental insurance status for U.S. military families. AB - This study explores what factors influence whether active duty U.S. military personnel enroll their families in Department of Defense (DoD) or non-DoD dental insurance plans. The data come from a 26-site, cross-sectional survey of U.S. service members conducted from April 1994 to January 1995. A prestratified, randomly selected target sample of 15,915 service members yielded 12,950 respondents (81% response rate); 7,243 of these had insurance-eligible families. Age, gender, ethnicity, education, rank, marital status, branch of service, number of children, number of years of military service, and insurance status of respondents were collected on self-administered questionnaires. We performed stepwise, backward, logistic regression analysis to determine which factors influence a military family's dental insurance status. Results show that enrollment in DoD insurance is influenced by every demographic factor collected; enrollment in non-DoD insurance is influenced by fewer factors. The decision by U.S. service members to enroll their families in dental insurance plans is subject to many and complex influences. PMID- 9866361 TI - Comparison of a portable capillary whole blood coagulation monitor and standard laboratory methods for determining international normalized ratio. AB - The international normalized ratio (INR) is the current standard for monitoring anticoagulation therapy. Although simple to determine, it normally requires venipuncture and extensive laboratory resources for specimen handling and analysis. The portable capillary whole blood coagulation monitor is an alternative to laboratory venipuncture. Its promoted advantages are: it obtains a blood sample by finger-stick versus venipuncture; rapid turnaround time for results; resultant dosage adjustments (as appropriate) performed in minutes versus hours or days after testing; relative ease of use by nonlaboratory personnel; and potential for home monitoring. This project compared the results of INRs obtained through the venipuncture/laboratory process to INRs obtained by the portable monitoring process at the National Naval Medical Center. A correlation coefficient of 0.97 was determined. The difference in the mean INR results of the two testing methods was not clinically significant (p = 0.269). The portable monitor was determined to be a viable alternative to laboratory testing. PMID- 9866363 TI - Mental health attrition from Air Force basic military training. AB - The Department of Defense invests considerable money in recruiting and training new military recruits who are discharged for mental health reasons before completing their first enlistment, often within the first 6 months of active duty. The purpose of this study was to provide a description of 1,138 Air Force recruits referred for a mental health evaluation while in basic military training. Variables examined include dispositions, demographic features, diagnoses, mental health history, and attitude and motivation characteristics. Findings show that trainees recommended for discharge often had a history of depression, expressed a lack of motivation to continue in the military, were reporting suicidal ideation, and typically had withheld information on their mental health history during their Military Entrance Processing Station processing. The results are discussed in terms of potential strategies for reducing mental health attrition in Air Force basic military training. PMID- 9866362 TI - Dual agency, dual relationships, boundary crossings, and associated boundary violations: a survey of military and civilian psychiatrists. AB - We hypothesized that psychiatrists with high dual-agency potential (military and health maintenance organization [HMO] psychiatrists) were more likely than non HMO civilian psychiatrists to engage in dual relationships, report pressures to do so, participate in other general boundary-crossing activities, and report associated counter-therapeutic outcomes (boundary violations). Ninety military and 191 demographically matched civilian psychiatrists reported the number of boundary-crossing activities (including dual relationships) and associated counter-therapeutic outcomes in the preceding year with adult patients. Military and HMO psychiatrists reported greater external pressures than non-HMO civilian psychiatrists to engage in dual relationships; however, all three groups were similar in their reported numbers of dual relationships. The reported boundary crossing activities and dual relationships studied here are not necessarily associated with reported boundary violations. The relative risk of a particular boundary crossing associating with harm to a patient likely depends on the therapeutic context and should be determined on a case-by-case basis. PMID- 9866364 TI - Evaluation of a bed utilization system in a surgical nursing section. AB - This research study evaluated a new bed utilization program established in the surgical nursing section at a major military medical facility. The study involved four wards with 83 beds before the new system was implemented, and three wards with 71 beds after the implementation. Five variables were identified as having an effect on the bed management program. These variables were time of discharge, escort compliance, bed availability, staff satisfaction, and patient satisfaction. Both a paired t test and an independent t test were applied for statistical analysis at the 0.05 level of significance. Data were collected for 1 month at two different intervals for comparison, using two surveys, patient records, 24-hour nursing reports, and same-day surgical schedules. A sample size of 103 was used as a basis for time of discharge and staff and patient satisfaction. The staff satisfaction survey also included a demographic data sheet. Escort compliance and bed availability were based on a 30-day time frame. The sample subjects included all patients who entered the surgical nursing section and their health care providers. Data analysis revealed significant differences in escort compliance and bed availability between the pre and post data. Findings from this study support the need for a functional, efficient bed management system. PMID- 9866365 TI - Early pulsatile lavage for the decontamination of combat wounds: historical review and point proposal. AB - Pulsatile lavage is an effective means of irrigation for grossly contaminated wounds. Pulse lavage was first used by oral surgeons in the U.S. Army during the war in Vietnam, and U.S. military investigators pioneered its early development and scientifically validated its efficacy. Modern pulse lavage units are ideally suited for military use because they enable first- and second-echelon medical personnel (including nonphysicians) to rapidly and effectively decontaminate combat wounds with minimal logistic burden. Pulse lavage units should become standard issue in all combat medical supply canisters or Authorized Medical Allowance blocks, and medical personnel should be trained in their use. PMID- 9866366 TI - Spine and spinal cord war injuries during the war in Croatia. AB - OBJECTIVE: The present report summarizes the experience of an evacuation hospital in southern Croatia in treating 96 patients with spine and spinal cord war injuries. METHODS: A retrospective review was done for 96 wounded persons (86 soldiers, 10 civilians) with spinal cord injuries from August 1991 through December 1995. The ages ranged from 15 to 59 years (mean, 28.3 years for soldiers, 38.5 years for civilians). Diagnostic procedures were plain radiography, computed tomography, and computed tomographic myelography. However, in most cases a more conservative surgical approach was used. RESULTS: The highest rates of admittance were recorded in 1992 (N = 38) and 1993 (N = 47). The lumbar spine was injured in 55% of the patients, the cervical and thoracic spines in 17.7%. All injuries were caused by projectiles from automatic rifles and sniper fire (51%) and from fragments of explosive devices (49%). Blast injury of the spinal cord was found in 10 patients. The most frequent complications caused by the fragments were wound infection, urinary tract infection, decubitus, and pneumonia. Four patients (4.2%) died in the hospital, and 43.0% of patients survived but were severely handicapped. CONCLUSION: Careful clinical examination combined with modern diagnostic imaging and use of broad-spectrum antibiotics reduced the need for surgical intervention in patients with spinal cord injuries. PMID- 9866367 TI - Changes in pulse rate among the civilian population during air-raid alerts in the city of Zagreb. AB - This investigation was performed in Zagreb and its surroundings in 1991 and 1992 during the war against Croatia. The study included 50 civilians who, at the moment of the sounding of the air-raid siren, had a continuous electrocardiogram recorded as part of the routine cardiologic examination. The frequency of the pulse was read out from the electrocardiogram at four different times: before the sounding of the siren, the moment of the sounding of the siren, the moment of cessation of the alert, and 1 hour afterward. The mean values of the pulse were calculated for each time point. The subjects were divided into four groups according to the length of exposure to the air-raid alerts (September 15, 1991 to January 3, 1992). The results show a significant increase in pulse frequency at the moment of the sounding of the air-raid siren compared with before the air raid siren (p < 0.001). However, no significant difference was found between the beginning and the end of air-raid alert campaign (p > 0.05). We conclude that within the civilian population of the city of Zagreb there was no adaptation to traumatic war situations as indicated by a lack of pulse reaction after a period of time under air-raid alerts. In other words, civilians always reacted with increased pulse frequency at the moment of the sounding of the air-raid siren. PMID- 9866368 TI - Endoscopic ultrasound localization of a pancreatic insulinoma: case report and review of the localization techniques. AB - A 23-year-old male was referred to our hospital for evaluation of new-onset seizures. Signs and symptoms of neuroglycopenia, including weakness, dizziness, and confusion, appeared during fasting and resolved promptly with intravenous dextrose administration. Insulin, proinsulin, and C-peptide levels were consistent with a diagnosis of insulinoma. Screening tests for multiple endocrine neoplasia type 1 and surreptitious sulfonylurea uses were negative. Preoperative localization of the insulinoma by transabdominal ultrasonography, computed tomography, and indium-111 octreoscanning were unsuccessful. Endoscopic ultrasonography (EUS) identified a 6- to 7-mm tumor at the juncture of the head and body of the pancreas. Surgical exploration confirmed the preoperative localization, and an 8-mm tumor was simply enucleated. The patient has been free of symptoms for 18 months since surgery. This report describes the utility of EUS to localize a solitary pancreatic insulinoma and provides a comparison of EUS and other preoperative localization techniques. PMID- 9866369 TI - Bilateral carotid artery dissection during the Army physical fitness test. PMID- 9866370 TI - Hypertensive left ventricular hypertrophy with left ventricular outflow tract obstructive physiology: a case of successful treatment with dual-chamber pacing. AB - Controversy exists over the use of atrioventricular synchronous pacing for patients with hypertrophic obstructive cardiomyopathy who are refractory to medical treatment. We present the case of an elderly man with severe-effort dyspnea attributable to hypertensive left ventricular hypertrophy with dynamic aortic outflow obstruction. We were able to achieve dramatic subjective and objective improvement in the exercise capacity of this patient with dual-chamber pacing after attempts at medical management had failed. This patient's response to therapy and his impressive response to minor pacemaker adjustments suggest that atrioventricular synchronous pacing should be considered for patients with secondary left ventricular hypertrophy as well as patients with hypertrophic obstructive cardiomyopathy. PMID- 9866371 TI - Choosing a family physician. What do patients want to know? AB - What type of information do patients find valuable when choosing a family physician? At a suburban Midwestern clinic, 221 adults rated the value of 12 demographic items and eight attributes pertaining to physicians. Of the demographic information, board certification was most valued by respondents; all personal attributes listed were rated very highly. Although 44% of women preferred a female physician, 93% of women agreed that, in general, male and female family physicians are equally competent. In summary, a physician's personal attributes and characteristics are of most importance to consumers when choosing a family physician. With the exception of board certification status, information about a physician's training or demographics is of much less value to patients. In addition, the information identified as most important in the survey is not typically provided by managed care organizations. PMID- 9866372 TI - Childhood cancer incidence and trends in Minnesota, 1988-1994. AB - Childhood cancer incidence patterns for Minnesota, obtained from the Minnesota Cancer Surveillance System, were compared with national rates as well as with historic data from eight Minnesota counties. In total, 1,140 neoplasms were diagnosed in children (ages 0 to 14) between 1988 and 1994. Leukemias were the most common diagnosis for boys (30.3%) and girls (29.6%), followed by central nervous system tumors. The average annual age-adjusted incidence rates for all cancer sites were 167.2 and 136.2 per million for boys and girls, respectively. These rates were somewhat higher than national rates. In particular, the incidence rate for astrocytoma in boys was significantly elevated. Childhood cancer incidence, particularly brain tumors, has increased in the eight-county region from 1969 to 1994. This analysis demonstrated the Minnesota's childhood cancer incidence patterns are similar to national patterns. PMID- 9866373 TI - MMA resolutions to AMA I-98 include smoking ban. PMID- 9866374 TI - A community approach to prenatal care. PMID- 9866375 TI - [Epidemiological study of intrapelvic venous congestion syndrome (IVCS) using new IVCS symptom score]. AB - BACKGROUND: The pathophysiology of prostatodynia still remains unknown. Recently, it was reported using three dimentional magnetic resonance venography (3D-MRV) that intrapelvic venous congestion (IVC) was found predominantly in patients with prostatodynia, and was designated this condition as IVC syndrome (IVCS). The present study was designed to reveal the prevalence of IVCS in taki drivers and office workers. METHODS: A total of 605 men (584 drivers and 21 office workers) in a taxi company in Kyoto city was enrolled in this study. They completed the IVCS symptom score, which was developed originally, including 5 questions (pain or discomfort at 1: the urethra during voiding, 2 lower abdomen, 3: perineal region, 4: inguinal region, 5: during ejaculation). The score of each question was graded from 0 to 5 points, and a total symptom score was obtained by adding these 5 scores to give a range of 0 to 25 points. 3D-MRV was performed to evaluate IVC. The diagnosis of IVC was determined based on the findings of the dilation of the prostatic capsular vein and/or the venous plexus of the lateral ligament of the bladder. RESULTS: Out of 603 men, 494 (81.9%), 40 (6.6%), 30 (5.0%) and 39 (6.5%) had a total symptom score of 0,1,2 and 3 or more. 3D-MRV was performed in 16 symptomatic men (scores of 3 or more), demonstrating IVC in 7 of them (43.8%). These 7 men were considered to have IVCS. In contrast, IVC was recognized in only one (6.3%) of 16 asymptomatic men (score of 0), were selected at random from 494 men scored 0 (p < 0.05). CONCLUSION: IVCS symptom score could be used as a screening tool for the detection of IVCS. PMID- 9866376 TI - [Predictability of Gleason score and reduction time (tau) of prostatic volume after castration for the prognosis of prostatic cancer]. AB - PURPOSE: The efficacy of the reduction time (tau) after castration as a prognostic factor was examined by comparing to Gleason score. MATERIALS AND METHODS: The change of prostatic volume after castration was observed from the castration to 3 months after in 24 cases of prostatic cancer. Prostatic volume was examined by transrectal ultrasonography of the prostate. Survival curves was calculated by Kaplan-Meier method. Differences among survival curves were analyzed using Cox-Mantel test. RESULTS: tau had a close relationship to the prognosis of each case (Wilcoxon test: p < 0.05, Cox-Mantel test: p < 0.05). Gleason score had a weak relationship to prognosis (Wilcoxon test: N. S., Cox Mantel test: p < 0.05). CONCLUSIONS: tau was efficient as prognostic factor compared to Gleason score. PMID- 9866377 TI - [Clinicopathological study on patients with renal cell carcinoma according to whole area histological sections]. AB - BACKGROUND: We tried to establish new classification of histological grade and indication of elective nephron-sparing surgery (ENSS) in renal cell carcinoma (RCC), and studied histological changes according to tumor size. METHODS: We made whole area histological sections on 142 cases with RCC and investigated histological aspects and prognosis. RESULTS: (1) To classify the grade of the cases, the worst grade which occupied more than 10% of the tumor provided most appropriate prognosis. (2) In 144 cases (kidneys), satellite tumor lesions (STL) were observed in 69 cases (47.9%), and the cases with vein invasion were observed in 75 cases (51.4%). Incidence of these two factors increased with tumor size. (3) From the point of view of location of STL and vein invasion, ENSS was possible with taking more than 2 cm surgical margin in the cases with less than 4 cm in size and slow growing type. But indication of ENSS should not be decided with ease. (4) Incidence of the cases with multiple structural and cell types and grades increased with tumor size. (5) Incidence of the cases with solid structure, spindle or pleomorphic cell type and high grade increased with tumor size. (6) It was supposed that tumor heterogeneity and progression could be associated with tumor growth in each cases. CONCLUSION: These results suggest the necessity of treatment of the cases with RCC as small as possible. If the tumor is less than 4 cm, prognosis tends to be good, and ENSS might be possible in some cases. PMID- 9866379 TI - [A case-control study on the efficacy and the optimum interval of mass screening for prostatic cancer]. AB - PURPOSE: The efficacy and the optimum interval of mass screening for prostatic cancer were investigated by a case-control study. MATERIALS AND METHODS: A matched pairs analysis by 1:5 was conducted between 31 cases of advanced prostatic cancer in stage C and D detected by mass screening and 155 controls of normal subjects selected at random. In all cases and controls, the history of taking part of the mass screening for last 3 years, then the estimates of odds ratios (OR) and 95% confidence intervals (95% CI) were calculated on discordant pairs of the matched analysis. RESULTS: OR to suffer from advanced prostatic cancer was 0.22 (95% CI: 0.07-0.70) in the group to submit to mass screening 1 year ago in comparison with the group not to submit once for 3 years. According to the analysis on the interval of submission to mass screening, the effect of screening to prevent the progression to advanced stages might be valied for 1 year. CONCLUSION: Mass screening for prostatic cancer every year might be efficient to reduce the number of patients progressing to advanced stages. PMID- 9866378 TI - [Clinical features of transurethral anterior sphincterotomy and urological management of patients with cervical spinal cord injury]. AB - BACKGROUND: The ideal urological management for the patients with cervical spinal cord injury (CSCI) is to obtain catheter free urination and to prevent urinary tract complications. We have evaluated cases that had undergone transurethral anterior sphincterotomy from the view-point of the operative indications and the efficacy. METHODS: We carried out sphincterotomy 166 times on 133 male patients with CSCI in our Center. Before the operations were performed, all patients suffered from urinary incontinence, and they were unable to catheterize themselves for low level activity of daily life. Before and after the operation, their detrusor functions with sphincter reactions were assessed by urodynamic study. In principle we have followed up these cases by cystogram combined with cystometry, cystogram and excretory-pyelography or abdominal ultrasonography. RESULTS: In post-operative evaluations, more than 80% of cases attained hypotonic detrusor contractions and residual urine was significantly decreased. In long term follow-up, 96% of patients had obtained catheter free urination and about 85% of patients had no urinary tract complications, such as bladder deformity, vesicoureteral reflux, or hydronephrosis, with the exception of common unavoidable urinary infections. About 20% of cases had to be re-operated upon, and some cases showed hypertonic detrusor contractions or detrusor-sphincter dyssynnergia during follow-up. CONCLUSION: The operative indications of sphincterotomy should be decided when the CSCI patients is unable to perform self catheterization, and when due to the dysfunction of the urethral sphincter, these patients suffered from voiding difficulties or autonomic dysreflexia, or when the urinary tract complications might occur. In the majority of cases the aims of the sphincterotomy were achieved but some cases underwent another operation or had recurrent dysfunction of the urethral sphincter, indicating the need for careful follow-up. PMID- 9866380 TI - [Preliminary experience of radical retropubic prostatectomy using an endoscopic stapler]. AB - PURPOSE: Preliminary experience of radical retropubic prostatectomy using an endoscopic stapler is reported. METHODS: An endoscopic stapler was applied for ligation and division of the lateral prostatic ligaments and the deep dorsal vein complex during radical retropubic prostatectomy in 8 patients. RESULTS: Procedures with stapler were easily performed and almost always effective for hemostasis. Mean total blood loss was 663 ml, mean 575 ml of autologous blood was given. None of patients was transfused allogeneic blood. CONCLUSION: These results indicate that an endoscopic stapler may facilitate radical retropubic prostatectomy. PMID- 9866381 TI - [Malignant mesenchymoma in the scrotum. A case report]. AB - A 67-year-old man visited our hospital with the chief complaint of painless swelling of the left scrotal content. An elastic hard mass was palpable in the left scrotum. Resection of the tumor, including the left scrotal skin, was performed. The tumor originated from the scrotal wall and did not communicate with the testis, the epididymis or spermatic cord. Histologically, the tumor was diagnosed as a malignant mesenchymoma composed of liposarcoma, chondrosacoma and osteosarcoma. Malignant mesenchymoma is very rare, especially in the scrotum. Our case that originated from the scrotal wall is the first one reported in Japan. PMID- 9866382 TI - [A juxtaglomerular cell tumor. Analysis of immunohistochemistry, electron microscopy and in situ hybridization]. AB - We present a case of juxtaglomerular cell tumor measuring 8 mm in diameter in the right kidney. The hypertension was cured and plasma renin activity returned to normal level following tumor resection with partial nephrectomy. We studied histopathologic, electron microscopic, and immunohistochemical findings of the tumor. The majority of tumor cells stores renin granules in cytoplasm. In situ hybridization confirmed us that the most tumor cells produce renin. We use the digoxigenin labeled 0.6-kb length RNA probe of human renal renin, the specificity was analyzed by competition assay (1:50). PMID- 9866383 TI - [A case of papillary cystadenoma of the epididymis]. AB - We report a case of papillary cystadenoma of the right epididymis in a 48-year old man. He consulted our hospital complaining of a small, painless mass at the upper pole of the right testis. There did not seem to be any remarkable problem of the mass. At 52 years old, the mass caused him pain, we diagnosed this case as chronic epididymitis, and performed right epididymectomy. On cut section, the tumor was multicystic and well encapsulated. The fluid within the cystic spaces was hemorrhagic. Microscopically, multimicrocysts were covered by tumor cells, which did not show any atypism or mitotic features. A papillary growth pattern was formed by vacuolated tumor cells around the capillaries in the microcysts, and a diagnosis of papillary cystadenoma of the epididymis was made. Papillary cystadenoma of the epididymis is considered a comparatively uncommon disease. To our knowledge, about 50 cases have been reported in the world literature, and this is the 17th case in Japan. Histologically, this tumor is similar in appearance to well-differentiated renal cell carcinoma, so it should be differentiated from metastatic renal cell carcinoma and multicystic papillary adenocarcinoma of the rete testis. The patient remains well with no evidence of recurrence or complication 8 months after excision. There is no evidence of renal cell carcinoma, nor features of von Hippel-Lindau's disease in this patient or his family. PMID- 9866384 TI - [Tuberculosis morbidity trends among children, adolescents and young adults in Uzbekistan]. AB - After a declining period by the mid-1980s and its stabilization in 1985-1995, the morbidity rates of tuberculosis in Uzbekistan increased in 1996 by 20.7% as compared to 1995, by 24% among children in the past decade, and 2 times among adolescents and young individuals aged under 30 years within 5 years. The rise of mortality rates is accompanied by aggravation of the clinical forms of the tuberculosis cases detected. The goal-oriented examinations of populations at risk of the disease in 14 districts of the Republic indicated a 1.5-fold increase in total mortality rates, followed by an upward tendency. PMID- 9866385 TI - [Organization of care for tuberculosis children who had contact with maternal hospital worker suffering from bacillar tuberculosis]. AB - While contacting a patient with bacillar tuberculosis, neonates and their mother are at risk for the disease, which makes it necessary to make up a programme that involves emergency antituberculosis measures. Among them primary specific drug therapy in all children in contacts, which may prevent tuberculosis in them, is highly effective. PMID- 9866386 TI - [Tuberculosis epidemiology-epizoology relationship in West Siberia]. AB - Comparison of tuberculosis epizootic tension with some parameters characterizing the epidemiological situation in areas with different incidence rates of cattle tuberculosis under present conditions showed their inverse correlation. It is suggested that with a drastic improvement of the epizootic situation, the cattle becomes an agent of natural immunization of inhabitants in poor districts, as evidenced not only by the lack of new cases of tuberculosis in human beings, but by an increase in cases with residual posttuberculous changes in the lung (14.1% of the stock breeders examined in the long poor tuberculosis area versus 3.7% in this infection-free area), as well as by high infection rates (74.2 versus 43.5%). PMID- 9866387 TI - [Social characteristics of patients in penitentiaries++]. AB - Three hundred and forty five convicted patients with tuberculosis underwent sociohygienic examinations. Among the examinees, the disease was first detected in 74.2% in a reformatory, in 25.8% in an investigating cell. The persons who fell ill with tuberculosis in places of confinement were mainly young, their ages ranged from 20 to 39 years. Only every 4 patients with tuberculosis are condemned for the first time, more than half the patients have over 3 convictions and every 5 patients have over 5 convictions. The most socially disadapted individuals fall ill with tuberculosis in the reformatory; among the examinees, 7.6% were homeless before conviction. The overwhelming majority of the convicts had no social contacts: 52.2% were never married. The study ascertained that in addition to the negative social environment, some criminogenic, personal, and behavioral factors of this contingent predispose to tuberculosis. PMID- 9866388 TI - [Special comprehensive inter-department program for tuberculosis control in the Cheliabinsk region]. AB - The special comprehensive multidisciplinary programme for tuberculosis control was used as a basis for planning. The developed special comprehensive interdepartmental programme for tuberculosis control in the region is aimed at involving all services and departments in whom tuberculosis control should be a national, official, professional duty. A multidisciplinary approach to organizing and making antituberculous measures is especially required as antituberculous institutions have the shortest funds now. The programme has approved by the regional administration. PMID- 9866389 TI - [Principles in differential diagnosis of female genital tuberculosis]. AB - The paper deals with current methods of diagnosis of female genital tuberculosis, which is very difficult despite numerous studies, it shows the frequency of its latent forms. The present classification of the disease and its clinical manifestations are given. The paper shows that only a complex of studies may make its diagnosis and determine the intensity of the process. It analyzes the results of tuberculin tests, microbiological, histological, X-ray studies. The paper shows it necessary to use the present diagnostic techniques, including laparoscopy and hysteroscopy. PMID- 9866390 TI - [Recovered reproductive function in female patient with sever genital tuberculosis]. AB - In severe genitoperitonial tuberculosis, conglomerate forms of the disease, and liquid accumulation between intestinal loop adhesions, laparotomy may verify the diagnosis in the shortest time while other diagnostic methods cannot be used due to contraindications for them. The operation should be maximally saving and only diagnostic. Timely combined antituberculous treatment concurrently with surgical interventions yields good results of recovered reproductive function. PMID- 9866391 TI - [Analysing effectiveness of traditional treatment for urogenital tuberculosis by outpatients follow-up]. AB - The paper presents the social and clinical structure of patients with urogenital tuberculosis through studies of 518 outpatient case histories. It has been ascertained that there is a prevalence of common and complicated forms of nephrotuberculosis, 82.4% of patients undergo surgery. Taking into account the fact that a fifth of patients cannot have long-term inpatient treatment, the incidence of Mycobacteriuria (67.9%), the prevalence of the disease, requiring organ-saving operations, the high degree of patients' disability, methods for intensifying urinary tuberculosis should be developed. PMID- 9866392 TI - [Clinical picture, course and therapeutic efficiency of pulmonary tuberculosis in patients with organochlorine pesticides and hepatitis B markers]. AB - A total of 356 patients with active forms of pulmonary tuberculosis were examined. Toxicological and radioimmunological (RIA) studies revealed serum organochlorine pesticides (OCP) and virus hepatitis B markers (VBHM) in 225 (63.2%) and 219 (61.6%), respectively. Pulmonary tuberculosis was demonstrated to run worse in patients with OCP and VBHM who tolerated antibiotics worse, showed a high incidence of side effects, lower efficiency of treatment. Pathogenetic benzonal therapy with enterosorption in tuberculosis enhances therapeutical efficiency by 20-25% and reduces the incidence of side effects by 1.5-2 times. PMID- 9866393 TI - [Current pathomorphism and specific features of complex tuberculous uveitis treatment]. AB - The study has indicated that in the 1990th, there has been considerable changes in the clinical picture and course of ocular tuberculous lesions. Their morbidity has tended to increase in children, adolescents, and young individuals. Patients with tuberculous uveitis with posterior eyeball lesions are more than 70%. The specific inflammatory process is primarily exudative and infiltrative, followed by rapid development of complications that prevail over the major focus, prevent treatment and substantially deteriorate visual functions. The findings are important not only for understanding the pathogenetic development of an ocular tuberculous process and they provide evidence for the approach to be used in the treatment of various manifestations of ocular tuberculosis. PMID- 9866394 TI - [Radiologic diagnosis of osteoarticular tuberculosis and borderline skeletal diseases]. AB - A total of 3305 ill children and adults treated for skeletal diseases and lesions underwent primary and follow-up radiation studies. Patients with spinal diseases amounted to 72%, of them 58% were found to have neurological diseases, 28% had articular diseases. The examination of patients involved X-ray tomography, contrast myelotomography, computed tomography, and magnetic resonance imaging. Surgical and laboratory verifications of the diagnosis were made in 53% of adults and 78.5% of children. It is shown that there are difficulties in diagnosing and differentiating osteoarticular tuberculosis at present and X-ray tomography and contrast myelography are essential in diagnosing spinal diseases. Indications for computed tomography and magnetic resonance imaging, osteoscintigraphy and the sequence of their use in the diagnosis of diseases were defined. The paper emphasizes that an osteoarticulogist's assessment of complex data of radiation diagnosis is very important. PMID- 9866395 TI - [Treatment policy for children with complicated osteoarticular tuberculosis]. AB - Experience in treating 97 children aged 1 to 14 years who had complicated tuberculosis of the spine (n = 74) and large joints (n = 23) in a specialized surgical clinical setting is presented. The developed treatment policy involves early radical reparative or reconstructive surgical intervention and intensive long-term therapy. The treatment allows one to cure 96.3% of children, yielding good functional results in 82.5% of them. PMID- 9866396 TI - [Surgical treatment of tuberculous spondylitis using ceramic implants]. AB - The use of osseous grafts in the treatment of tuberculous spondylitis is associated with substantial shortcomings, requiring a long-term bedside regimen, followed by fixation of the spine with a rigid corset. The application of ceramic implants allowed one to attack several tasks: to provide primary stabilization of the vertebral segment, to exclude corset fixation, to reduce a postoperative period and surgical traumatism by refusing to take an autograft. Forty nine operations were performed in patients aged 23-65 years. In the early postoperative period neurological disorders completely disappeared in 7 patients, the remaining has significant positive changes. Complete osteoceramic adhesion occurred 2-4 months later. Following 2 patients, 26 patients were stricken off the disability register. The remaining patients continued to be disabled (7 and 16 patients had disability groups 2 and 3, respectively) due to significant contaminant diseases. PMID- 9866397 TI - [Experimental rationale and clinical evaluation of perichondroplasty used in surgery for tuberculosis and nonspecific joint diseases]. AB - Experiments on 77 rabbits studied the specific features of chondrogenesis during different grafting modifications for the perichondrium. The latter was found to have a high chondroplastic potential. The factors predisposing to cartilaginification and inhibiting this process were identified. Experimental positive results of perichondrial arthroplasty, in tuberculous arthritis as well, were achieved. By taking into account the experimental data, original methods for reparative operations used in patients with tuberculosis and nonspecific diseases of the joints were developed in the clinical setting. Their use in 12 patients yielded positive results, suggesting that the above methods are promising and their further study is advisable. PMID- 9866398 TI - [Specific features of DNA isolation for polymerase chain reaction in extrapulmonary tuberculosis]. AB - While testing samples by the polymerase chain reaction (PCR), a procedure for DNA isolation is advisable adapted for different types of materials; thorough DNA purification is of the greatest importance in testing blood-containing samples, deep sample lysis is of particular importance for osseous tissue samples. A differential approach to isolating DNA from clinical samples allows the efficiency of PCR to be enhanced. PMID- 9866399 TI - [Use of blast transformation test with different tuberculin dilutions to evaluate functional activity and directivity vector of T-cell activation in patients with tuberculosis during chemotherapy]. AB - The lymphocyte blast transformation test (LBTT) with three tuberculin dilutions was used to examine 190 patients with varying pulmonary tuberculosis activity, of them 63 patients received chemotherapy. According to the blast formation in the patients' cultured peripheral blood cells by three tuberculin dilutions, a correlation was found between the clinical manifestations of the process and the functional activity of T lymphocytes. Thus, the greatest percentage (500 TU) of blasts in LBTT per mean PPD dose was detectable in patients with low LBTT results by three tuberculin dilutions with positive dynamics during chemotherapy. With further positive dynamics, the proportion of blasts in the cultured peripheral blood cells was highest per high PPD doses (5000 TU). On the contrary, patients with progressive tuberculosis displayed a oppositely directed phasic pattern. PMID- 9866400 TI - [Immunity of middle age and aged patients with tuberculosis and its changes during multimodality treatment by using T-activin]. AB - Fifty six patients with pulmonary tuberculosis were examined. Of them 30 had T activin in multimodality treatment. To supplement the agent in elderly and old patients was found to elevate the counts of T lymphocytes and T helper cells, to normalize Tx/Tc ratios, to enhance T lymphocytic activity and IL-2 synthesis. At the same time there was an increase in nonspecific responsiveness: enhancement of NKC activity and IL-1 synthesis by macrophages. With this, stimulation of specific antituberculous immunity took place. Normalization of immunological responsiveness and stimulation of specific antituberculous immunity were associated with enhanced therapeutical efficiency. PMID- 9866401 TI - [Assessment of homeostasis by body's adaptative responses in patients with tuberculous pleurisy]. AB - The types of adaptative responses were defined, by analysing leukograms in 126 patients with tuberculous pleurisy. Assessing homeostasis by proteinograms, cellular and humoral immunity verified adaptative responses as an integral index of responsiveness. This identified 5 types of responsiveness. Abnormal (hyperactive, hyporeactive, paradoxical, and areactive) responsiveness was found in 78.6% of patients with tuberculous pleurisy. All outcomes with significant residual pleural changes were observed in patients with abnormal responsiveness. Adaptative responses may be used in the clinical setting not only for evaluation of homeostasis, but for prediction of prognosis and for organization of controlled pathogenetic therapy on an individual basis. PMID- 9866402 TI - [Oxidative metabolic changes, antioxidantion, and platelet activation rat phagocytes during development of experimental specific tuberculous changes]. AB - An experiment was made on 100 noninbred albino rats, of which 80 rats were intraperitoneally inoculated by Mycobacterium tuberculosis (MT) in a dose of 7.5 mg. Examinations were conducted 1 day, 1, 2, and 8 weeks after inoculation. Alveolar macrophages, nonfractionated cellular sediment of bronchoalveolar lavage, and leukocytes were the object of the studies. Spontaneous and BCG stimulated HCT test, the activities of superoxide dismutase (SOD) and catalase, the content of malonic dialdehyde (MDA), and platelet activation factor (PAF) were determined in all cell populations. The course of the process was histologically controlled. Following a fortnight, specific foci developed in the organs, they began spontaneously resolving 6 weeks later. In all types of cells, an infectious process resulted in an increase in the rate of oxidative metabolism, which did not lead to their functional exhaustion. In early infection, the activity of SOD dropped, the level of MDA and the activity of catalase increased. During involution of specific changes, their normalization of MDA and catalase variables corresponded to the high values of SPD in the cells. The level of PAF moderately elevated during the formation of specific changes in the organs and fell below the control values in the involutional phase of the process. PMID- 9866403 TI - [Normal lungs and mediastinum X-ray in axial computed tomograms]. PMID- 9866404 TI - [Pleural puncture biopsy in pleurisies of unknown etiology]. PMID- 9866405 TI - [Spontaneous pneumomediastinum]. AB - The authors present 7 cases with spontaneous pneumomediastinum (SPM), a rare pathological condition. The cause of SPM is rupture of marginal alveoles and spread of air into the mediastinum through perivesical and peribronchial fats. Males are more frequently affected, but females were prevalent in this study (as 4:3). At the stage when air does not spread outside the mediastinum, Hamman's sign, i.e. "crepitation" ("a crunching sound", "a noise of air bursting bubbles") heard at cardiac auscultation is significant in the diagnosis. Dissection of air in all mediastinial structures, which is detectable during X-ray studies both at this stage of SPM, and at the stage of spread of air to the neck, chest, and abdomen is an important diagnostic sign. The evolution of SPM is generally good and conservative therapy leads to recovery in most patients. In 9-16% of patients, the evolution of the process is dramatic and requires cervical mediastinotomy after Tiegel or thoracotomy with wide mediastinotomy. PMID- 9866406 TI - [Combination of disseminated tuberculosis and cancer of lung]. PMID- 9866407 TI - [Elevated amylase in pleural malignant effusions]. PMID- 9866408 TI - [Contribution of Doppler ultrasonography to the study of natural history of portal hypertension]. PMID- 9866409 TI - Influence of the smoking habit in the surgery of inflammatory bowel disease. AB - The smoking habit is a key factor in the development of inflammatory bowel disease (IBD), but little information exists as to the relationship between smoking habit, the need of surgery and its complications. OBJECTIVES: To investigate the relationship between smoking habit, the need of surgery, their complications and clinical recurrence after surgery in Crohn's disease (CD) and ulcerative colitis (UC). METHODS: We studied a group of 62 patients (22 with UC and 40 with CD) with previous surgery. We analyzed the clinical and surgical characteristics of the disease. Smoking habit was established by a personal interview. This group of patients was compared with another control group of 202 patients (133 with UC and 69 with CD) with IBD without previous surgery. RESULTS: Smoking habit was similar between operated and non-operated patients for both UC (73% and 80% non-smokers) and CD (67% and 63% smokers) The number and type of complications after surgery were not related with smoking habit. In CD patients, although the recurrences did not depend on the smoking habit, they did occur earlier in smokers than in non smokers (83.6 +/- 21 vs 155 +/- 50 weeks, p = ns). CONCLUSIONS: The smoking habit does not seem to influence significantly the need of surgery and post surgical development of IBD, although in CD the smokers seems to present recurrence before non smokers. PMID- 9866410 TI - Hidrocolonic ultrasonography in the detection of tumoral processes in the inferior gastrointestinal tract. AB - OBJECTIVES: To determine the value of hidrocolonic ultrasound in the detection of proliferative lesions in the colon and to compare it with other techniques of already proven value. MATERIAL AND METHODS: We performed a prospective blinded trial including 155 patients (82 males and 73 females) with ages ranging from 33 to 94 years (average of 58) and clinical and analytical criteria suggesting the existence of colonic proliferative lesions. Patients with rectal mass or those with deficient bowel preparation were excluded. Ultrasound findings were compared to those obtained by colonoscopy (133 cases) and by Barium RX studies (22 cases) and all diagnoses were always confirmed by histologic exams. RESULTS: 155 patients were studied. 50 of them had cancer and 46 of these 50 were diagnosed by Hidrocolonic Ultrasound (92%); 19 had polyps > 7 mm. and 15 of these (78.9%) were diagnosed by Hidrocolonic Ultrasound. Hidrocolonic Ultrasound failed to detect all the polyps < 7 mm. The overall sensitivity, specificity, positive predictive value and negative predictive value for identifying colon carcinoma were 92%, 98%, 95.8% and 96.2% respectively and for polyps > 7 mm were 78.9%, 100%, 100% and 97.1%, respectively. The mean time for examination was 14 minutes. Tolerance was good in 114 patients (73.5%), 29 showed a slight discomfort (18.7%) and 12 (7.7%) showed a great discomfort. There were no complications. CONCLUSIONS: Hidrocolonic ultrasound is an innocuous, fast, well tolerated technique for detecting colonic proliferative lesions > 7 mm H.U. can be considered as a useful complementary technique to other more expensive and invasive ones, such as barium RX studies and Colonoscopy. PMID- 9866411 TI - Surgical treatment of the acute cholecystitis in the laparoscopic age. A comparative study: laparoscopy against laparatomy. AB - OBJECTIVE: The aim of this study was to assess the complications and results of the laparoscopic opposite to open treatment of the acute cholecystitis. METHODS: A retrospective randomized study with two groups of 30 patients each one. The parameters tested were age, sex, risk factors, surgical time, hospital stay, cholecystitis type, and early or late complications. RESULTS: In the two groups there were no significant differences in age, sex, risk factors, type of cholecystitis and surgical time. The average of hospital stay was significantly longer for open cholecystectomy (9.5) than for laparoscopic technique (2.30) (p < 0.001). The complication rate was higher (7.30%) in open cholecystectomy. CONCLUSIONS: The laparoscopic cholecystectomy should be the standard procedure for the treatment of the acute cholecystitis. PMID- 9866413 TI - [Usefulness of ultrasonography in the diagnosis of portal hypertension]. AB - Portal hypertension syndrome is a common evolutive complication of several hepatic and extrahepatic diseases, being liver cirrhosis responsible for more than 80% of cases. When diagnosed it has prognostic value because of the high incidence of hemorrhagic, metabolic and infectious complications that these patients may develop. Clinical suspicion must be confirmed by objective complementary studies that provide information about the etiology and severity of the disease. In this review article we describe the contribution of ultrasonography in the evaluation of patients with portal hypertension as an objective diagnostic method and the usefulness of doppler ultrasound in the non invasive hemodynamic assessment of the splanchnic and portal circulation. PMID- 9866412 TI - Anorectal function in patients with complete rectal prolapse. Differences between continent and incontinent individuals. AB - AIMS: A study is made of the alterations in anorectal physiology among rectal prolapse patients, evaluating the differences between fecal continent and incontinent individuals. PATIENTS AND METHODS: Eighteen patients with complete rectal prolapse were divided into two groups: Group A (8 continent individuals) and Group B (10 incontinent women), while 22 healthy women were used as controls (Group C). Clinical exploration and perineal level measurements were performed, along with anorectal manometry, electrophysiology, and anorectal sensitivity to electrical stimuli. RESULTS: The main antecedents of the continent subjects were excess straining efforts, while the incontinent women presented excess straining and complex deliveries. Pathological perineal descent was a frequent finding in both groups, with a hypotonic anal canal at rest (p < 0.001 vs controls) and at voluntary squeezing (p < 0.001 vs controls). In turn, the incontinent patients exhibited a significantly lower anal canal pressure at rest than the continent women (p < 0.05). There were no significant differences between Groups A and C in terms of pudendal motor latency, though latency was significantly longer in Group B than in the controls (p < 0.01). Moreover, pudendal neuropathy was more common, severe and often bilateral in Group B. There were no differences in rectal sensation to distention or in terms of the volumes required to relax the internal anal sphincter. In turn, both prolapse groups exhibited diminished anal canal and rectal sensitivity to electrical stimuli. CONCLUSIONS: Patients with rectal prolapse exhibit a hypotonic anal canal at rest, regardless of whether they are continent to feces or not. Continent patients have less pudendal neuropathy and therefore less pressure alterations at voluntary sphincter squeeze than incontinent individuals. PMID- 9866414 TI - [Budd-Chiari syndrome: current medical-surgical management]. AB - The Budd-Chiari syndrome or obstruction of the hepatic veins and/or suprahepatic inferior vena cava is a rare process, frequently associated with hypercoagulable states. There exist several clinical presentations, being most common the acute and subacute forms and rarely seen the fulminant and chronic forms (cirrhosis or fibrosis associated). We present here a female patient with Budd-Chiari syndrome produced by polycythemia, resolved by mesentericocaval interposition "H" shunt using internal jugular vein, after analyzing the current different modalities of medical-surgical treatment, specially portal system shunts and liver transplantation, related to presentation form. PMID- 9866415 TI - [Intestinal obstruction caused by solitary colonic metastasis of breast carcinoma]. PMID- 9866416 TI - [Late diagnosis of post-traumatic diaphragmatic hernia]. PMID- 9866417 TI - [Diagnostic errors in hiatal hernia]. PMID- 9866418 TI - [Apoptosis of peripheral leukocytes in patients with myelodysplastic syndromes]. AB - Apoptosis of peripheral leukocytes in 9 patients with myelodysplastic syndromes (MDS) was examined in vitro, using peripheral blood that had been gently incubated at 37 degrees C for 5 hours. The MDS patients included 3 with refractory anemia, 2 with refractory anemia with ringed sideroblasts, and 4 with refractory anemia with excess blasts. Peripheral blood specimens were also obtained from a control group consisting of 10 patients with iron deficiency anemia (IDA), 10 with idiopathic thrombocytopenic purpura (ITP) and 10 healthy individuals. Apoptotic granulocytes (Apo-Gs) were identified by morphological changes, including nuclear fragmentation, and expressed as a percentage of every 300 granulocytes counted. Apo-Gs were counted 1, 2, 3, 4, and 5 hours after incubation. Although the percentage of apo-Gs climbed over time in the MDS patients, a small number of apo-Gs were also observed in the healthy individuals. In the MDS patients, the proportions of apo-Gs 5 hours after incubation (37 degrees C) were significantly higher than those in the IDA and ITP patients and healthy individuals (15.7 +/- 8.0% in MDS patients vs. 2.8 +/- 1.2% in IDA patients, 2.3 +/- 1.7% in ITP patients, and 0.7 +/- 0.6% in healthy individuals; p < 0.005). No significant differences were observed in the proportions of apo lymphocytes. DNA fragments were observed in blood lymphocyte from an MDS patient examined. Negative correlations between the percentages of granulocytes and Apo Gs tended to be observed in the MDS patients. These results suggest that a strong susceptibility to peripheral granulocyte apoptosis is one of possible causes of granulocytopenia in MDS patients. PMID- 9866419 TI - [Allogeneic peripheral blood stem cell transplantation in 30 patients with hematologic disorders]. AB - Thirty patients (median age of 32 years; range, 6-61) with hematologic disorders received unmanipulated peripheral blood stem cell transplants from HLA-matched or one-antigen-mismatched related donors following myeloablative therapy for acute lymphoblastic leukemia (7), acute myelogenous leukemia (6), chronic myelogenous leukemia (8), myelodysplastic syndrome (3), or other disorders (6). Granulocyte colony stimulating factor (G-CSF) mobilized peripheral blood stem cells were collected from donors in 1 to 3 aphereses. The apheresis products contained mean counts of 11.3 x 10(8) (range, 3.8-17.2) nucleated cells/kg and 6.7 x 10(6) (range, 1.3-16.7) CD34+ cells/kg. Graft-versus-host-disease (GVHD) prophylaxis consisted of cyclosporin A plus methotrexate, or FK506 plus methotrexate. All patients received G-CSF following their transplant. Although 1 patient died of pneumonia 6 days after transplantation, the others demonstrated rapid engraftment. Median days to recovery to 500/microliter neutrophils and 20,000/microliter platelets were 13 (range, 8-21) and 14 (range, 1-23) days, respectively. The incidence of acute GVHD grade II-IV was 33%; chronic GVHD developed in 57% of the assessable patients. There were no episodes of graft failure or rejection. Nineteen patients (63%) were alive and in complete remission from 147 to 839 days following their transplant (median follow-up of 560 days). Further follow-up study will be required to assess the incidence of chronic GVHD and graft-versus-leukemia (GVL) effects. PMID- 9866420 TI - [Studies of childhood non-Hodgkin's lymphoma--treatment results with the CCLSG NHL 960 protocol. Children's Cancer and Leukemia Study Group (CCLSG)]. AB - We report here on the preliminary treatment findings of a CCLSG NHL 960 study that was initiated in March 1996. In this study, 37 patients with non-Hodgkin's lymphoma were assigned to 4 different treatment groups according to disease stage and histology: (1) localized disease; (2) advanced disease, lymphoblastic type; (3) advanced disease, large cell type; and (4) advanced disease, Burkitt type. The first three groups received the modified protocols of the NHL 890 study. Groups 1 and 3 received COPADM induction therapy (CPM, VCR, PRD, ADR, and MTX). After achieving remission, Group 1 received only maintenance therapy consisting of alternate administration of 7 drugs, while Group 3 received additional intensification therapy with combination chemotherapy consisting of MTX and Ara C, followed by a maintenance phase involving the administration of 9 drugs. Group 2 received COPADL induction therapy (CPM, VCR, PRD, ADR, and LASP) and consolidation/intensification therapies followed by a maintenance phase. Group 4 received short-term intensive COPADM polychemotherapy. Twelve patients with localized with localized disease (stage I-II) and 25 patients with advanced disease (stage III-IV) were enrolled in this study. Except for 2 patients in the advanced disease stages who died earlier in the course of the study, all patients remained in remission. PMID- 9866421 TI - [The frequency and development of tissue iron deficiency in 6 iron deficiency anemia patients with plummer-vinson syndrome]. AB - The physical signs of tissue iron deficiency include smooth and red tongue, angular stomatitis, koilonychia, and pica. The incidence of these conditions is unknown in Japan. We evaluated the frequency and development of tissue iron deficiency in 353 patients with iron deficiency anemia. The frequency of tissue iron deficiency was 6.8%; papillary atrophy of the tongue, 5.4%; abnormal nails, 5.4%; angular stomatitis, 1.1%; Plummer-Vinson syndrome, 1.7%; and pica, 0.06%. These findings were compared with the date collected by Wintrobe and Beveridge. The development and incidence of tissue iron deficiency correlated significantly with the severity of iron deficiency anemia. PMID- 9866422 TI - [Systemic aspergillosis caused by an aflatoxin-producing strain of Aspergillus in a post-bone marrow transplant patient with acute myeloid leukemia]. AB - We report a case in which an aflatoxin-producing strain of Aspergillus flavus (A. flavus) caused systemic aspergillosis in a post-transplant 41-year-old man with acute myeloid leukemia. The leukemia was initially resistant to two courses of induction chemotherapy. The third course of chemotherapy, however, induced complete remission. Thereafter, the patient underwent bone marrow transplantation from his HLA identical brother. Pulmonary aspergillosis was suspected as a complication during induction chemotherapy. Twenty days after the transplant, the patient's absolute neutrophil count had increased to 500/microliter. However, the symptoms of pulmonary aspergillosis were aggravated following neutrophil and monocyte recovery. The patient died of sinus arrest due to complete atrioventricular block 31 days after his transplant. At autopsy, we found that the fungus had invaded the brain, lungs, spleen, kidneys, skin, and myocardium, including the sinoatrial conduction system. There was no sign of acute graft versus-host disease. A strain of A. flavus was isolated from cultured tissue samples of fungal lesions and shown by thin-layer chromatography to produce aflatoxins. To our knowledge, this is the first case report describing an infection by an aflatoxin-producing A. flavus. PMID- 9866423 TI - [Immunoblastic lymphadenopathy-like T cell lymphoma with high levels of serum interleukin-6, cold agglutinin disease, and immune thrombocytopenia]. AB - An 88-year-old woman was admitted with generalized lymphadenopathy, anemia, and thrombocytopenia. On admission, a peripheral blood examination showed a red blood cell count of 146 x 10(6)/microliter, a hemoglobin concentration of 6.9 g/dl, and a platelet count of 5.0 x 10(4)/microliter. Blood examination detected polyclonal hypergammaglobulinemia; the results of the direct/indirect Coombs' test were positive; and an elevated cold agglutinin titer and high platelet associated IgG (PA-IgG) level indicated the existence of autoantibodies. Serum cytokine measurements disclosed an elevated level of interleukin-6 (IL-6). Immunoblastic lymphadenopathy-like T cell lymphoma was diagnosed on the basis of lymph node biopsy specimens. VP-16 and steroid therapy alleviated the patient's lymphadenopathy, anemia, thrombocytopenia, and hypergammaglobulinemia. These findings suggest that tumor cells with a T cell phenotype produced IL-6 in large quantities, thus provoking B-cell and plasmacytic histologic changes and humoral disease manifestations, including hypergammaglobulinemia. PMID- 9866424 TI - [Cytarabine-induced pericarditis]. AB - We report two cases of cytarabine (Ara-C) induced pericarditis. The first patient was a 61-year-old man with de novo acute myelogenous leukemia (AML) with trilineage myelodysplasia (M2). Fever and pericardial effusion developed after treatment with low-dose Ara-C and etoposide (VP16) during the patient's second leukemic relapse. The second patient was a 65-year-old man with myelodysplastic syndrome (MDS) transformed into AML (M2). He achieved a complete remission after treatment with low-dose Ara-C and VP16. However, during consolidation chemotherapy consisting of oral cytarabine ocfosfate and VP16, fever and pericardial effusion developed. In both cases, pericardiocentesis revealed exudative effusion with many neutrophils. Neither bacterial nor fungal infection was demonstrated. Chest symptoms and fever were resolved by methylprednisolone pulse therapy and pericardial drainage. Both patients were given a diagnosis of pericarditis caused by allergic reaction to Ara-C. We also report on 6 other patients with Ara-C syndrome who have been encountered by our hospital. PMID- 9866426 TI - [Macrocytic anemia with anisocytosis due to alcohol abuse and vitamin B6 deficiency]. AB - Megaloblastic anemia due to folic acid deficiency and ringed sideroblastic anemia have been reported in alcohol abusers. It has also been reported that vitamin B6 deficiency causes ringed sideroblastic anemia as well as microcytic anemia that is not associated with ringed sideroblasts. We encountered a case of macrocytic anemia with anisocytosis in a 75-year-old alcohol abuser who suffered vitamin B6 deficiency. Neither megaloblastic changes nor ringed sideroblasts were observed in specimens of the patient's bone marrow. Analyses of porphyrin content and heme biosynthetic enzyme activity suggested a decline in ALA-synthase activity (an enzyme that depends on vitamin B6) as well as decreased ferrochelatase activity or abnormal iron metabolism. Abstention from alcohol led to a reduction in mean corpuscular volume and the disappearance of Pappenheimer bodies commonly observed in the red blood cells of drinkers. Follow-up supplements of vitamin B6 resolved the patient's anisocytosis and anemia. PMID- 9866425 TI - [Successful treatment with high dose chemotherapy following peripheral blood stem cell transplantation for relapsed Burkitt's lymphoma with central nervous system involvement]. AB - A 19-year-old man complaining of right upper abdominal pain underwent surgery for the removal of a testicular tumor in October 1997. He was given a diagnosis of Burkitt's lymphoma and was in clinical stage II B. The patient went into completed remission with VABCOP-M combination chemotherapy, but relapsed with involvement of the central nervous system (CNS). He was treated with ICE, then CHASE together with total cranial irradiation and simultaneous intrathecal MTX and cytosine arabinoside through Ommaya reservoir until a second remission was achieved. Afterward, the patient was given high-dose chemotherapy and total body irradiation followed by an autologous peripheral blood stem cell transplant (auto PBSCT), and maintained complete remission. Though the prognosis for Burkitt's lymphoma with CNS involvement is considered to be poor, high-dose chemotherapy with PBSCT was a successful treatment for relapsed Burkitt's lymphoma in our patient. PMID- 9866427 TI - [Successful treatment with CPT-11 and adriamycin for hemophagocytic syndrome associated with intravascular lymphomatosis]. AB - A 75-year-old man was admitted because of non-Hodgkin's lymphoma (histology undetermined) of the rib. A complete remission was achieved after CHOP therapy and irradiation. One year later, high fever, thrombocytopenia and liver dysfunction developed. Bone marrow aspirate revealed a hypoplastic marrow with hemophagocytic histiocytes, and a diagnosis of hemophagocytic syndrome (HPS) was made. Although no lymphomatous lesions were detected, HPS due to relapsed lymphoma was strongly suspected. The patient received MEVP therapy including etoposide and prednisolone, but without any improvement. Soon after the initiation of CPT-11 and adriamycin (ADM) therapy, all symptoms of HPS disappeared. This combination chemotherapy was repeated over a three-week span, and the patient remained in partial remission for the next 10 months. In November 1997, a tumor developed in the paranasal sinus, and the patient died three months later. The autopsy disclosed many B lymphoma cells filling the small vessels of almost all organs, and a final diagnosis of intravascular lymphomatosis (IVL) was made. These findings indicate that combination CPT-11 and ADM therapy is effective for cases of IVL accompanied by HPS that are refractory to conventional chemotherapies. PMID- 9866428 TI - [Effective splenectomy in a patient with HIV-associated thrombocytopenia]. AB - A 31-year-old man presented with a 3-month history of petechial hemorrhages. Physical examination revealed no splenomegaly. The patient's platelet count was 1.0 x 10(9)/l and bone marrow aspiration showed an elevated number of megakaryocytes. A diagnosis of HIV-associated thrombocytopenia was made on the basis of HIV seropositive results. The CD4 cell count was 400 x 10(6)/l. No opportunistic infections indicating AIDS were detected. Initially the patient was treated with predonisolone, but showed only a transient response. He also failed to respond to zidovudine, lamivudine, or indinavir. Following splenectomy, however, his platelet count rose above 80 x 10(9)/l (normal level: 150-350 x 10(9)/l). PMID- 9866429 TI - Ambulatory and inpatient procedures in the United States, 1996. AB - OBJECTIVES: This report presents estimates of surgical and nonsurgical procedures performed in the United States during 1996. Data are presented by characteristics of patients, region of the country, and procedure categories for ambulatory and inpatient procedures separately and combined. METHODS: Estimates in this report are based on data collected from the National Hospital Discharge Survey (NHDS) and the National Survey of Ambulatory Surgery-(NSAS). NHDS provides data on hospital inpatient care, and NSAS provides data on ambulatory surgery in hospitals and in freestanding ambulatory surgery centers. For NHDS, data were collected for approximately 282,000 discharges from 480 non-Federal short-stay hospitals (95 percent response rate). For NSAS, data were collected for approximately 125,000 ambulatory surgery discharges from 488 hospitals and freestanding ambulatory surgery centers (81 percent response rate). RESULTS: An estimated 71.9 million procedures were performed on 39.9 million discharges from hospitals and freestanding ambulatory surgery centers during 1996: 40.4 million procedures were for inpatients, and 31.5 million were for ambulatory patients. Females had more procedures than males, and the rate of procedures increased with age in ambulatory and inpatient settings. The leading procedures for ambulatory surgery patients and inpatients combined were arteriography and angiocardiography, endoscopy of small intestine, endoscopy of large intestine, and extraction of lens. PMID- 9866430 TI - Patient attitudes about mandatory reporting of domestic violence. Implications for health care professionals. AB - As of January 1994, California physicians are required to report to police all patients who are suspected to be victims of domestic violence. This article describes the results from a focus group study of abused women (n = 51) that explored their experiences with and perspectives on medical care. The eight focus groups included two Latina (total n = 14), two Asian (total n = 14), two African American (total n = 9), and two Caucasian (total n = 14) groups of women who had been the victims of domestic abuse within the previous 2 years. The women were recruited through community-based organizations in the San Francisco Bay Area. With regard to physician reporting of domestic violence to police, five themes were identified: fear of retaliation by the abuser, fear of family separation, mistrust of the legal system, desire for police protection, and preference for confidentiality and autonomy in the patient-health professional relationship. Our results indicate that mandatory reporting may pose a threat to the safety and well-being of abused women and may create barriers to their seeking help and communicating with health care professionals about domestic violence. PMID- 9866431 TI - Occupational asthma. Practical points for diagnosis and management. AB - Asthma is a common chronic illness characterized by episodes of reversible airflow obstruction. A cornerstone of asthma management is identifying and avoiding agents that cause bronchospasm. The workplace is an important potential source of respirable exposures that can cause or trigger asthma. Identification of an occupational factor in asthma is important: early diagnosis and removal of the worker from the exposure is associated with improved prognosis; the diagnosis of occupational asthma may lead to compensation for work-related impairment and disability; and the diagnosis of occupational asthma is a Sentinel Health Event with implications for public health and prevention. In this article, we review specific causes of occupational asthma and general settings in which an occupational factor should be suspected and explored as part of the management of the worker with asthma. We also review specific and simple elements of history and pulmonary function testing that can be easily assessed by most health care practitioners and may be sufficient to establish a diagnosis of occupational asthma. Finally, we review the medical-legal implications of occupational asthma. PMID- 9866433 TI - A randomized double-blind study of the effect of distant healing in a population with advanced AIDS. Report of a small scale study. AB - Various forms of distant healing (DH), including prayer and "psychic healing," are widely practiced, but insufficient formal research has been done to indicate whether such efforts actually affect health. We report on a double-blind randomized trial of DH in 40 patients with advanced AIDS. Subjects were pair matched for age, CD4+ count, and number of AIDS-defining illnesses and randomly selected to either 10 weeks of DH treatment or a control group. DH treatment was performed by self-identified healers representing many different healing and spiritual traditions. Healers were located throughout the United States during the study, and subjects and healers never met. Subjects were assessed by psychometric testing and blood draw at enrollment and followed for 6 months. At 6 months, a blind medical chart review found that treatment subjects acquired significantly fewer new AIDS-defining illnesses (0.1 versus 0.6 per patient, P = 0.04), had lower illness severity (severity score 0.8 versus 2.65, P = 0.03), and required significantly fewer doctor visits (9.2 versus 13.0, P = 0.01), fewer hospitalizations (0.15 versus 0.6, P = 0.04), and fewer days of hospitalization (0.5 versus 3.4, P = 0.04). Treated subjects also showed significantly improved mood compared with controls (Profile of Mood States score -26 versus 14, P = 0.02). There were no significant differences in CD4+ counts. These data support the possibility of a DH effect in AIDS and suggest the value of further research. PMID- 9866432 TI - Controversies in perioperative medicine. AB - Coexisting medical problems are common in surgical patients. Due to the lack of convincing studies, however, the optimal strategies for perioperative management of many medical problems remain unclear. Thus, practices are often based on inconclusive clinical studies or extrapolated from current understanding of pathophysiology. This article reviews guidelines and practices for endocarditis prophylaxis, perioperative anticoagulation in patients with mechanical heart valves, glucose control in diabetic patients undergoing surgery, and the use of beta-blockers to prevent postoperative cardiac complications. In addition, the strength of the evidence supporting these practices is evaluated. PMID- 9866435 TI - New vaccines for infectious diseases. PMID- 9866436 TI - New tests for detecting HIV infection. PMID- 9866434 TI - Prevalence of antimicrobial drug-resistant Streptococcus pneumoniae in Washington State. AB - We conducted a survey to assess the prevalence and geographic distribution of antimicrobial drug resistance among invasive isolates of Streptococcus pneumoniae in Washington State. Sequential sterile-site pneumococcal isolates were submitted from 13 hospital laboratories between 1 October 1995 and 30 January 1997. We serotyped 275 isolates from adults and children and determined minimum inhibitory concentrations (MIC) for commonly used antimicrobial drugs. Data were abstracted from medical records to compare differences in outcome and risk factors for infection. Of the 275 isolates, 73 (26.5%) were nonsusceptible to one or more antimicrobial drugs. Penicillin-nonsusceptible pneumococci (PNSP, MIC > or = 0.1 microgram/ml) accounted for 42 (15.3%) of the 275 isolates including 4 (1.5%) resistant strains (MIC > or = 2 micrograms/ml). The 42 PNSP included serogroups 6, 9, 14, 19, and 23, all of which are represented in the 23-valent pneumococcal vaccine. PNSP were also nonsusceptible to trimethoprim/sulfamethoxazole (92.9%), erythromycin (38.1%), imipenem (28.6%), and ceftriaxone (23.8%). Forty-seven (17.1%) of the 275 isolates were multiple drug-nonsusceptible pneumococci (MDNSP). A significantly greater number of patients < or = 12 years of age were infected with MDNSP compared with those > 12 years. Prior use of antimicrobial drugs and an immunosuppressive disorder were risk factors for infection with PNSP. In summary, pneumococci nonsusceptible to penicillin and other antimicrobial drugs are prevalent among adults with invasive pneumococcal disease in Washington State. A large proportion of PNSP are resistant to other commonly used antimicrobial drugs. Local antibiotic susceptibility data should be considered when designing empiric treatment regimens. PMID- 9866437 TI - Hepatitis C. PMID- 9866438 TI - Indoor environmental quality. PMID- 9866439 TI - Update on public health in correctional facilities. PMID- 9866440 TI - Acute effects of exposure to methyl tert-butyl ether in gasoline. PMID- 9866441 TI - Motor vehicle occupant protection for children. PMID- 9866442 TI - Asthma--the shift from episodic treatment to ongoing prevention, education, and management. PMID- 9866443 TI - Influenza. PMID- 9866444 TI - Mushroom poisoning due to amatoxin. Northern California, Winter 1996-1997. PMID- 9866445 TI - Thrombophilia in ischemic stroke. PMID- 9866446 TI - Acquired sulfhemoglobinemia. An underreported diagnosis? PMID- 9866447 TI - Shuffling off this mortal coil. A Shakespearean perspective on death and dying. PMID- 9866448 TI - Modeling reflex asymmetries with implicit delay differential equations. AB - Neuromuscular reflexes with time-delayed negative feedback, such as the pupil light reflex, have different rates depending on the direction of movement. This asymmetry is modeled by an implicit first-order delay differential equation in which the value of the rate constant depends on the direction of movement. Stability analyses are presented for the cases when the rate is: (1) an increasing and (2) a decreasing function of the direction of movement. It is shown that the stability of equilibria in these dynamical systems depends on whether the rate constant is a decreasing or increasing function. In particular, when the asymmetry has the shape of an increasing step function, it is possible to have stability which is independent of the value of the time delay or the steepness (i.e., gain) of the negative feedback. PMID- 9866449 TI - Modeling actin filament reorganization in endothelial cells subjected to cyclic stretch. AB - Hemodynamic forces affect endothelial cell morphology and function. In particular, circumferential cyclic stretch of blood vessels, due to pressure changes during the cardiac cycle, is known to affect the endothelial cell shape, mediating the alignment of the cells in the direction perpendicular to stretch. This change in cell shape proceeds a drastic reorganization at the internal level. The cellular scaffolding, mainly composed of actin filaments, reorganize in the direction which later becomes the cell's long axis. How this external mechanical stimulus is 'sensed' and transduced into the cell is still unknown. Here, we develop a mathematical model depicting the dynamics of actin filaments, and the influence of the cyclic stretch of the substratum based on the experimental evidence that external stimuli may be transduced inside the cell via transmembrane proteins which are coupled with actin filaments on the cytoplasmic side. Based on this view, we investigate two approaches describing the formulation of the transduction mechanisms involving the coupling between filaments and the membrane proteins. As a result, we find that the mechanical stimulus could cause the experimentally observed reorganization of the entire cytoskeleton simply by altering the dynamics of the filaments connected with the integral membrane proteins, as described in our model. Comparison of our results with previous studies of cytoskeletal dynamics reveals that the cytoskeleton, which, in the absence of the effect of stretch would maintain its isotropic distribution, slowly aligns with the precise direction set by the external stimulus. It is found that even a feeble stimulus, coupled with a strong internal dynamics, is sufficient to align actin filaments perpendicular to the direction of stretch. PMID- 9866450 TI - A Bayes-optimal sequence-structure theory that unifies protein sequence-structure recognition and alignment. AB - A rigorous Bayesian analysis is presented that unifies protein sequence-structure alignment and recognition. Given a sequence, explicit formulae are derived to select (1) its globally most probable core structure from a structure library; (2) its globally most probable alignment to a given core structure; (3) its most probable joint core structure and alignment chosen globally across the entire library; and (4) its most probable individual segments, secondary structure, and super-secondary structures across the entire library. The computations involved are NP-hard in the general case (3D-3D). Fast exact recursions for the restricted sequence singleton-only (1D-3D) case are given. Conclusions include: (a) the most probable joint core structure and alignment is not necessarily the most probable alignment of the most probable core structure, but rather maximizes the product of core and alignment probabilities; (b) use of a sequence-independent linear or affine gap penalty may result in the highest-probability threading not having the lowest score; (c) selecting the most probable core structure from the library (core structure selection or fold recognition only) involves comparing probabilities summed over all possible alignments of the sequence to the core, and not comparing individual optimal (or near-optimal) sequence-structure alignments; and (d) assuming uninformative priors, core structure selection is equivalent to comparing the ratio of two global means. PMID- 9866451 TI - Selection dynamics in autocatalytic systems: templates replicating through binary ligation. AB - The theory of autocatalytic binary ligation is reviewed within the context of a consistently applied Michaelis-Menten quasi-steady-state approximation to obtain explicit analytical results describing time-course data from experiments. A detailed protocol for the step-wise elucidation of a minimal set of experimental parameters is outlined. The kinetic equations are then generalized to cases of self- and cross-catalysis among an arbitrary number of different templates and applied to experiments involving just two templates. Depending on the values of various kinetic parameters such systems can display exclusionary Darwinian selection corresponding to an exponential growth law, selective coexistence or coexistence of all species characteristic of a parabolic growth law; the intermediate behaviour arises as a property of the full mechanism analysed here. Our results are applicable to the classical case of self-replicating nucleic acids and their analogues as well as to newly discovered self-replicating peptides. PMID- 9866452 TI - Pulse vaccination strategy in the SIR epidemic model. AB - Theoretical results show that the measles 'pulse' vaccination strategy can be distinguished from the conventional strategies in leading to disease eradication at relatively low values of vaccination. Using the SIR epidemic model we showed that under a planned pulse vaccination regime the system converges to a stable solution with the number of infectious individuals equal to zero. We showed that pulse vaccination leads to epidemics eradication if certain conditions regarding the magnitude of vaccination proportion and on the period of the pulses are adhered to. Our theoretical results are confirmed by numerical simulations. The introduction of seasonal variation into the basic SIR model leads to periodic and chaotic dynamics of epidemics. We showed that under seasonal variation, in spite of the complex dynamics of the system, pulse vaccination still leads to epidemic eradication. We derived the conditions for epidemic eradication under various constraints and showed their dependence on the parameters of the epidemic. We compared effectiveness and cost of constant, pulse and mixed vaccination policies. PMID- 9866453 TI - Euthanasia and physician-assisted suicide revisited. PMID- 9866454 TI - Fluconazole sensitivities of Candida species isolated from the mouths of terminally ill cancer patients. AB - Oral candidosis is common in advanced cancer and is often treated with the systemic triazole antifungal drug fluconazole. This study examined the species of yeast present in the mouths of 30 patients with advanced cancer and determined their sensitivity to fluconazole. Thirty-five yeast isolates were collected from a total of 25 (83 percent) of the patients sampled. The two most common species were Candida albicans (15 isolates) and C. glabrata (11 isolates)--with smaller numbers of C. tropicalis, C. parapsilosis, C. guilliermondii, C. inconspicua, and Saccharomyces cerevisiae. The minimal inhibitory concentrations (MIC) of fluconazole for the strains of C. albicans were generally low (median 0.19 microgram/ml) but were considerably higher for C. glabrata (median 2 micrograms/ml). The remaining species demonstrated MICs similar to those for C. albicans, with the exceptions of C. inconspicua and Saccharomyces cerevisiae, which were relatively insensitive. In conclusion, non-albicans yeasts are common in the mouths of patients with advanced cancer and these may have reduced sensitivity to fluconazole. Mycological diagnosis is a valuable aid to management. PMID- 9866455 TI - The palliation of dyspnea in terminal disease. AB - Dyspnea is a complex subjective experience that is common in terminal illness. Patients may present at any time during the course of their illness, although prevalence increases with disease progression. Dyspnea has physical, psychological, social and spiritual components; without recognizing how each of these contributes to the total suffering of dyspnea, management is unlikely to be successful. The management of dyspnea involves both pharmacological and non pharmacological treatment. The main pharmacological palliative treatments are oxygen, opioids, and benzodiazepines, but the evidence to support these treatments is limited. More research is urgently needed to establish the efficacy of current treatments and to identify new ones. PMID- 9866456 TI - When oral morphine fails in cancer pain: the role of the alternative routes. PMID- 9866457 TI - Development of an outcome measure to document pain relief for home hospice patients: a collaboration between nursing education and practice. AB - In the spring of 1997, an Eastern Michigan University nursing educator, a master's nursing student, and a hospice clinical nursing director collaborated to develop a pain outcome instrument as a part of the student's clinical practicum. The instrument was designed to describe and measure pain control of home hospice patients in preparation for fall accreditation of a home hospice program by the Joint Commission on Accreditation of Healthcare Organizations. A chart audit of all on-call communications (n = 63 charts) by the educator and student supported the organization's need to improve patients' pain control. The collaborators reviewed the pain instrument literature, developed measurement instruments, and presented information to nurses at staff meetings for input and instrument revisions. A pilot study of the Home Hospice Patient Pain Assessment Instrument (HHPPAI) was conducted, the instrument was revised, and a second pilot was performed (n = 51 pain episodes). A description of those findings is presented. Further research needs to be conducted to examine instrument reliability and validity. The use of a pain outcome instrument, such as the HHPPAI, can improve patients' pain control and, therefore, promote their quality of life. PMID- 9866458 TI - Metastatic spread and common symptoms. Part One: Introduction, bladder cancer, and brain cancer. AB - This is the first of a multi-part series detailing the metastatic spread and natural history of 18 common tumors. The introduction summarizes (1) the concept of symptom/problem anticipation and early identification; (2) the process of cancer metastasis; and (3) the 18 tumors that each cause more than 6,000 deaths per year in the United States. Bladder cancer is discussed, with information included on tumor types, metastatic spread and invasion, and common symptoms. Obstructive uropathy is the oncologic emergency most commonly associated with bladder cancer. Sites of spread, resulting problems, and assessment parameters are presented as a table. Brain cancer (and brain metastasis) is discussed, with information included on tumor types, invasion (the primary means of spread in primary brain cancer), and common symptoms. Increased intracranial pressure and, to a lesser extent, spinal cord compression are the oncologic emergencies most commonly associated with brain cancer. Sites of spread, resulting problems (including site-specific symptoms), and assessment parameters are presented as a table. PMID- 9866459 TI - Can we afford to be passive in an aggressive environment? PMID- 9866460 TI - Review of mental health law must be courageous. PMID- 9866461 TI - NVQ care workers or licensed practical nurses? PMID- 9866462 TI - Interventions for confusion and dementia. 3: Reminiscence. AB - The aims of reality orientation, which was discussed in the previous article of this series (Vol 7(17): 1018-20), are to achieve behavioural improvements. The aims of reminiscence, however, are empowerment and the provision of pleasure and it is therefore more associated with quality of life and quality of care. Reminiscence can be an enjoyable activity. Used in a structured way, it can achieve a sense of wellbeing, and increase empathy between healthcare workers and clients. This article discusses the benefits and problems of reminiscence, and suggests means to develop reminiscence skills and materials. PMID- 9866464 TI - Synthetic casting tapes: benefits and uses of Delta-Cast Conformable. AB - Casting is a traditional practice that has changed relatively little over thousands of years. Today, however, with the emphasis on evidence-based medicine, health professionals are examining the practice and investigating ways of improving it for the benefit of both patients and those who perform it. Most fractures and many soft tissue injuries are usually managed in a rigid cast, which brings with it complications and disadvantages. The ideal form of fracture fixation would stabilize the fracture while allowing some movement of the fracture site and soft tissue compression. This article explores the use of Delta Cast Conformable (Johnson and Johnson), a synthetic casting tape, in both primary and secondary casting. The role of Delta-Cast Conformable with regard to focused rigidity casting, a new philosophy in fracture management, is discussed. PMID- 9866463 TI - Urinary catheters: selection, maintenance and nursing care. AB - Catheterization is often undertaken without sufficient assessment, and the high rates of bacteriuria (10-20%) occurring in catheterized patients seem to be accepted (Ward et al, 1997). This article reviews catheter selection, maintenance and nursing care, highlighting problem areas and suggesting measures that may be taken to reduce the risk factors. A problem-solving approach, based on research evidence relating to catheter care, is presented. Often there is a simple solution to a problem, but treatments based on traditions or myths appear to be the first choice in many situations. This article aims to provide nurses with the evidence base required to establish and maintain a urinary drainage system safely. PMID- 9866465 TI - Patient safety and the reuse of single-use medical items. AB - Trusts owe a duty of care to patients, staff, the public and the environment. This article considers the risk issues associated with the reuse of medical devices. The practice of reusing medical devices labelled by the manufacturer for single-use only must be carefully considered by managers and clinicians responsible for patient safety, to ensure that account has been taken of outcomes which may adversely affect clinical procedure and be harmful to the patient. PMID- 9866466 TI - How can the clinical credibility of nurse lecturers be improved?: 2. AB - The movement of nurse education into the university sector has put further tension on the theory-practice gap and the clinical credibility of nursing lecturers. If the ideal is for the lecturer to guide the learner in applying theory to practice and vice versa (Jarvis and Gibson, 1985), this can only realistically be achieved if lecturers of nursing have experience as practitioners within the field(s) they teach and are viewed as being clinically credible (ENB, 1996). In the first article (Vol 7(8): 490-2) an innovative way of enabling lecturers of nursing to engage in practice was presented. In this second article a variety of other approaches which enable lecturers to engage in clinical practice will be explored. PMID- 9866467 TI - Non-consensual sterilization of the adult with learning disabilities. AB - Community nurses may be asked for advice on sterilization operations for adults with learning disabilities by worried parents/carers. This article sets out the legal position advocated by the English courts. Sterilization for adults with learning disabilities is generally non-consensual. The courts cannot consent on behalf of the adult but can rule on the lawfulness of the operation. Cases need not be brought before the court when the operation is to be carried out to treat a specific menstrual malady and where sterilization is an incidental result. However, the Law Commission (1995) has set guidelines which recommend that such operations require a certificate from an independent medical practitioner. In operations where the sole purpose is contraception the courts will always need to be involved. Their decision on the lawfulness of the operation will be based on what is in the person's best interests which in turn will be determined by reference to standards set by a responsible body of medical practitioners. PMID- 9866468 TI - Nursing diagnosis in learning disabilities nursing. AB - The concept of nursing diagnosis and the relevance of individual diagnostic terms from the North American Nursing Diagnosis Association (NANDA) (1992) Classification Taxonomy 1-Revised are discussed in the context of learning disabilities nursing. The NANDA system of nursing diagnoses was implemented at an adult training centre with a client group who had a severe or profound learning disability. The diagnostic terms used to describe the phenomena found in everyday practice are identified. Due to the inadequacy of the NANDA term 'impaired verbal communication', a new nursing diagnosis, 'impaired non-verbal communication', is suggested. A method of recording nursing practice is used which shows client outcomes through diagnostic modifiers (a modifier denotes a change in the nursing diagnosis) over measured time periods. The value of a generic classification of diagnoses is demonstrated in a systematic approach to learning disabilities nursing practice. PMID- 9866469 TI - Acute surgical wound care. 2: The wound healing process. AB - The first article in this series on acute surgical wound care traced the history of surgical wound care from primitive dressings and techniques of closure used in the past to the present-day approaches. It also outlined the classification of acute surgical wounds (Vol 7(18): 1101-6). This second article describes the four stages of wound healing in acute surgical wounds, using clinical slides to illustrate the wound healing process. General factors, such as age, nutrition and medication, and local factors, including a moist environment, blood supply and wound infection, will be discussed to demonstrate their importance in promoting optimum wound healing. PMID- 9866470 TI - The blunders found in nursing documentation. PMID- 9866471 TI - The Bedouin way of life. PMID- 9866472 TI - The Internet--a new information medium for nurses--Part 1. AB - Part one of this series examined some Internet basics. Computer requirements and Internet connection options were covered, and Web addresses and search engines were reviewed. With this information in hand, you can determine if your current computer equipment is adequate or what type of computer and modern to purchase, and the type of Internet connection that will be right for you. Once online, remember to type web addresses exactly as they appear and to read the help sections of various search engines to learn how to perfect your search techniques. Part two of this series will cover practical issues of Internet use. Suggestions will be offered for dealing with e-mail file attachments, and how to avoid unsolicited e-mail and computer viruses. Techniques for reviewing and evaluating online health information will be discussed as well. PMID- 9866473 TI - Making cancer bearable: the Interlink Community Cancer Nurses model of supportive care. AB - Most cancer patients and their families experience the full impact of their diagnosis and the disruptions cancer brings to their lives in their own homes and communities. Access to supportive care programs within the community is critical if we are to assist these individuals in coping and adjusting to the realities of cancer. Interlink Community Cancer Nurses, a nonprofit nursing agency, provides supportive care across the continuum of cancer care from early diagnosis and treatment through to remission and/or palliative care through its independent model of supportive care delivered by expert oncology nurses. The program utilizes a unique, client-centred model of care delivery to ensure that needs are met, and to link patients to community services which promote their ability to meet the demands of cancer. Interlink's program of supportive care demonstrates that oncology nurses can make a significant impact on the outcomes of cancer for individuals and their families which may impact on cost, influence survival and enhance quality of life. PMID- 9866474 TI - Effectiveness and efficiency of nurse-given cancer patient education. AB - Increasing health care costs make it imperative that nurse-given cancer education programs be evaluated according to their economic effectiveness and efficiency. The purpose of this research was the comparison of two programs of nurse-given education at a Canadian cancer centre examining patients' knowledge of chemotherapy and associated side effects, the number of self-care behaviours for side effects, the degree of extra health resource utilization, patient and nurse satisfaction with teaching programs, and the nursing time needed for teaching. Results show that the experimental intervention is more effective and more efficient than the usual well-organized patient education program at this centre. These study findings begin to address the knowledge gap regarding the cost effectiveness of a critical component of ambulatory cancer nursing. PMID- 9866475 TI - The research experience: reflections of a novice researcher. AB - The oncology nurses at the Windsor Regional Cancer Centre (WRCC), like other health professionals, realize that professional proficiency can be improved through systematic strategies designed to enhance and streamline patient care. For this reason, all nurses in the systemic therapy department of the WRCC agreed to take part in a research study called "Effectiveness and Efficiency of Nurse Given Cancer Patient Education" (Porter, 1998). Their opinions, accomplishments and feelings about this research experience are recorded under selected headings of the research process. PMID- 9866476 TI - The bone marrow transplant experience through the eyes of a student nurse. PMID- 9866477 TI - The early detection of prostate cancer. Prostate Cancer Alliance of Canada. AB - In determining whether or not to undergo early detection tests (PSA and DRE), men must weight the possibility of early diagnosis and treatment of potentially aggressive prostate cancer against the limitation of these tests and decisions they will be faced with regarding treatment choices, effectiveness and side effects. The Prostate Cancer Alliance recommends that men 50 years of age or older talk to their physicians and inform themselves about the benefits and risks of early detection testing using PSA and DRE in order to make an informed decision about whether to have the tests. Men in higher risk categories (those with a family history of the disease or with an African Canadian ancestry) should consider this recommendation starting at age 40. Extensive information is available on these matters. Men should request such information from their family physician or their urologist and consult any or all of the groups sponsoring this message. PMID- 9866478 TI - High dose rate brachytherapy--a starting up your own program. PMID- 9866479 TI - Rebecca Bergman, RN, EdD, retired professor, University of Tel Aviv, Israel. Interview by Beverley Page. PMID- 9866480 TI - Alpha 2-agonist agents: a review for CRNAs. AB - Over 300 clonidine-related research articles have been prominently featured in anesthesia journals over the last 4 to 5 years. Nurse anesthetists may already find themselves administering alpha 2-adrenergic agents to help control hypertension; however, in the future their use as anesthetic adjuncts to control pain may also become important. This article reviews the alpha 2-adrenergic (A2) receptor and one of its agonist agents, clonidine. PMID- 9866481 TI - Nitric oxide: a primer for the practicing anesthetist. AB - Nitric oxide (NO) is receiving interest because of its use in patients with acute respiratory distress syndrome. Because more studies are being conducted, researchers are discovering a vast array of physiological effects attributable to NO. NO has been implicated as having a role in the endogenous regulation of blood pressure; smooth, skeletal, and cardiac muscle relaxation; renal and immune system functioning; and selective pulmonary vasodilation. This article looks at the pharmacological and physiological effects of NO and its current use in the clinical setting. PMID- 9866482 TI - Intrathecal opioids for the control of pain experienced during labor and in the post-cesarean section period. AB - Since the discovery of opiate receptors in the brain and spinal cord, considerable research has been performed to include intrathecal opioids for the control of pain. No area has used this knowledge more than the practice of obstetrical anesthesia. Intrathecal opioids have been shown to be very effective in controlling the pain experienced in the first stage of labor but have been ineffective in controlling second-stage labor pain. Intrathecal opioids, especially morphine sulfate, provide long-term analgesia but are associated with a high number of side effects. Research studies have attempted to determine the optimal dose of intrathecal opioids to provide the greatest amount of analgesia with the lowest incidence of side effects. Intrathecal morphine sulfate has been shown to be effective in controlling first-stage labor pain but has been shown to be the most efficacious in controlling the pain experienced in the post-cesarean section period. This article reviews the current literature and provides background of information to understand these developments. PMID- 9866483 TI - The effect of intrathecal and epidural narcotic analgesia on the length of labor. AB - A number of studies have shown conflicting results on the effect of epidural (EPI) analgesia on the length of labor. Combined spinal-epidural (CSE) and intrathecal analgesia (ITA) techniques have been used to provide pain relief for parturients, but currently there are few studies comparing EPI, CSE, and ITA techniques and their effect on the length of labor. Intrathecal opioids provide immediate pain relief for the parturient without autonomic, sensory, or motor blockade. The purpose of this retrospective study is to identify the effects of ITA on the length of labor compared with other regional anesthesia techniques. A total of 213 uncomplicated obstetric charts were reviewed in a 70-bed Air Force hospital that provided EPI, CSE, and ITA for obstetrical patients. The study consisted of the following four groups: (1) CSE (n = 76), (2) EPI(n = 41), (3) ITA (n = 49), and (4) no regional analgesia (NR) (n = 47). The results showed that the length of first-stage labor was statistically significantly less for those who received ITA compared with those who received CSE, EPI, and NR (P < .001). Second-stage labor was statistically significantly shorter for the NR group compared with the CSE and EPI groups (P < .01). No statistically significant difference was found between the length of second stage for ITA and NR groups. ITA analgesia shortened the first stage significantly in both primipara and multipara patients (P < .01). These results imply that the use of intrathecal opioids in the obstetrical patient does not prolong labor and seems to shorten the first stage of labor in both the primipara and multipara patients. PMID- 9866485 TI - Anesthetic drug interactions. Quarterly update. PMID- 9866484 TI - Pulmonary edema and hyponatremia during hysteroscopic resection of uterine fibroids: case report. AB - Hyponatremia and fluid overload--the so-called TURP Syndrome--is a rare but well understood complication of endoscopic resection of the prostate caused by rapid absorption of irrigating fluids from venous sinuses opened during the surgery. A similar syndrome occurring during endoscopic resection of the uterine mucosa has been postulated but only rarely described. New equipment and surgical techniques have recently been developed that allow for more invasive and extensive hysteroscopic surgery, which will inevitably lead to an increased incidence of this complication. We report the occurrence of a case of acute pulmonary edema occurring intraoperatively during hysteroscopic surgery, complicated postoperatively by severe hyponatremia. Anatomic and physiological causes of this syndrome are discussed, and recommendations are offered for avoiding and treating this potentially lethal complication. PMID- 9866486 TI - Issues and perspectives affecting CRNA practice. PMID- 9866487 TI - A guide to understanding informed consent. AB - Anesthesia providers are expected to provide information to the patient during the preanesthesia interview that enables the patient to make informed choices. Adequate disclosure during the informed consent process ensures the equalization of the practitioner/patient relationship and the decision-making rights of the patient. Both certified registered nurse anesthetists (CRNAs) and anesthesiologists are not only legally required to provide information that will allow a patient to make an informed judgment about how to proceed with various anesthetic modalities but are also obligated by their standards of practice. This article informs the CRNA about the principles of informed consent so that they can better understand their role in the informed consent process. PMID- 9866488 TI - US Food and Drug Administration: adverse event reporting. AB - This article reviews adverse event reports associated with anesthesia devices submitted to the US Food and Drug Administration during the period of August 15, 1896 to August 15, 1998. Cardiovascular, general surgical, and plastic surgical devices are the most frequently reported devices. Deaths are most frequently associated with cardiovascular, general hospital, and gastrourological devices. The most frequently reported failures associated with ventilators are failures of audio or visual alarm systems. PMID- 9866489 TI - Improving patient care by reporting problems with medical devices. AB - Healthcare practitioners are the primary users of medical devices for direct patient care. As such, they are in the best position to recognize problems that result from the use of medical devices. The outcome of a device-related adverse event or product problem, as with any other medical product, can be serious and result in illness injury, or even death. The sooner that FDA learns about a problem, the sooner the agency can take action to protect patient and user safety. Healthcare practitioners are major contributors to the knowledge base related to device use and safety through astute monitoring, rapid identification of device-related problems, and reporting these problems. An understanding of the voluntary and mandatory mechanism of reporting will ensure that device problems are reported appropriately and in a timely manner. As the primary users of medical equipment for direct patient care, health care professionals have the training and expertise to improve patient care by reporting actual and suspected problems with medical devices. PMID- 9866490 TI - The National Practitioner Data Bank: implications for nurse anesthetists. AB - The National Practitioner Data Bank (NPDB), created by the 1986 Health Care Quality Improvement Act, has been in operation since 1990. Hospitals and other credentialing bodies must query the NPDB when granting and renewing privileges. The NPDB receives about 25,000 reports of adverse actions against health practitioners each year. The NPDB was designed to be a flagging system providing information to licensing or credentialing authorities who would further examine practitioner records. Its purpose is to ensure that decision makers have information that might not otherwise be readily available, especially in the case of incompetent practitioners who move from hospital to hospital or state to state. Access to NPDB information is a concern for consumers and providers alike. Only 2% of matched reports to the NPDB made a difference in hospital privileging decisions. A limitation of NPDB information is that malpractice payments recorded in the NPDB do not necessarily constitute a comprehensive and definitive reflection of actual health care incompetence. All health care providers need to be aware of the NPDB, its mission, potential impact on their ability to be credentialed, and proposed additional uses of its information. PMID- 9866491 TI - Setting the record straight on nurse anesthesia and medical anesthesiology education. AB - The history, qualifications, capabilities, and legal status of nurse anesthetists in the United States have been perceived by organized anesthesiology as both a professional and economic threat to the medical specialty. Such threats often lead to turf battles in which groups try to seek public affirmation of their point of view through ongoing public and/or government relation debate and activism. Medicine, including anesthesiology, has used educational preparation of physician and nurse specialists as a favorite topic for such activism. In this nation's competitive environment, facts are often prone to distortion knowingly or unconsciously. This article is aimed at setting right the facts in the current debate used by the American Society of Anesthesiologists in regards to the comparative analysis of CRNA and anesthesiologist education. Because medicine most often uses length of education as a quality measure of that education, regardless of the validity of such arguments, this comparison is set within that framework. Unfortunately, it will not be the last work on this subject. PMID- 9866492 TI - Anesthetic drug interactions. Quarterly update. PMID- 9866493 TI - Evidence-based practice, research, peer review, and publication. AB - For about a quarter of a century, concerns have been expressed about published biomedical research. It became more acute after some published research and broad dissemination was found fraudulent. With the emphasis now being placed on scientifically validated or evidence-based practice, it has become more imperative that clinical guidelines be based on credible information in our textbooks and research literature. Since the early 1990s, it has been found that much of the research in our electronic databases does not meet quality standards and often is irrelevant, calling into questions problems with peer review, including the selection and publication process of our journals. This column is devoted to calling attention to these problems not only to CRNAs and other researchers, but also to the consumers of research who often use it to make changes in their practice. It also calls attention to the CRNA community about the movement toward calls for greater accountability in practice, both as to quality and cost, from which the movement toward evidence-based practice, the identification and benchmarking of best practices, and the development and implementation of clinical practice guideline has evolved. To feel ownership in anesthesia-related clinical practice guidelines, CRNAs must become involved in their development and implementation. PMID- 9866494 TI - Debate of the age. PMID- 9866495 TI - Drugs--who needs them? PMID- 9866496 TI - Clinical supervision: making it happen. AB - When owned and governed by those who implement it, clinical supervision can become a living, breathing part of professional working life, and not just another ritual. A multidisciplinary team from Southampton explains. PMID- 9866497 TI - Nursing and gerontology: a marriage of minds. AB - John Adams reports on a quiet revolution in educational support for nurses specialising in the care of older people as nursing aligns itself decisively with the rapidly expanding academic discipline of social gerontology. And for those new to the field, he offers ideas on where to access information. PMID- 9866498 TI - The construction of challenging behaviour. PMID- 9866500 TI - Elderly care takes a look at recent initiatives in skin care. PMID- 9866499 TI - Caring for the individual. AB - People do not lose their individuality as they grow older. With the use of a moving case history Carolyn Best describes how a lifetime's experiences and personality should be taken into account when dealing with older people. PMID- 9866501 TI - What makes a good nurse leader? PMID- 9866502 TI - Care within the family. PMID- 9866503 TI - Defining dementia. PMID- 9866504 TI - Make yourself heard. PMID- 9866505 TI - Nursing older people from ethnic minority communities. AB - As people from ethnic minority communities grow into old age, nurses must respond positively to their needs. A resource guide recently published by a working group established by the RCN's Race and Ethnicity sub-committee, urges nurses to both understand the different health needs of ethnic groups and to respect the involvement of older people in their own care. Here, we summarise the guide's main points. PMID- 9866506 TI - Community hospital-issues for older people. PMID- 9866507 TI - Taking care of the frail elder: a vital balance. PMID- 9866508 TI - A case report: special needs of hospitalized elders. AB - A case report is used to discuss common problems and complications of hospitalized elders. The report also illustrates the need for proactive hospital staff who are knowledgeable about the special needs of the aging population if this group is to be cared for successfully. The geriatric resource nurse model involves informal unit-based education aimed at expanding staff nurses' geriatric knowledge and is one of several models sponsored by the Hartford Foundation to improve care of the hospitalized elderly. Teaching rounds between geriatric resource nurses and advanced practice nurses are a primary feature of this model. PMID- 9866509 TI - Feeding the anorectic client: comfort foods and happy hour. AB - The anorectic patient presents a dilemma to the health care provider. As the patient refuses oral intake, quality of life diminishes, and health care costs increase. This article describes two approaches that have demonstrated effectiveness: comfort food and happy hour. "Comfort foods" describe foods associated with bygone years, intended to trigger recollections of pleasant childhood experiences and feelings of caring and healing. A second alternative described is the offering of a "happy hour" beverage, presented in a social milieu. These treatments are reproducible in a variety of clinical care settings and stimulate oral intake in the failure-to-thrive patient. PMID- 9866511 TI - Late-life drinking among women. AB - Although alcoholism exists among older people, the number of cases is unknown, and many alcoholics, particularly women, are unidentified. Late-onset alcoholism is a risk condition for older women. Nurses are in a strategic position to recognize this underidentified condition and to assist affected women in understanding what caused them to rely on alcohol. PMID- 9866510 TI - Current procedures for diagnosing dysphagia in elderly clients. AB - Intubation no longer remains the only solution to feeding problems of the elderly patient with dysphagia. Dysphagic disorders result from neurogenic, myogenic, psychogenic, or mechanical causes. Thus numerous hospitalized or institutionalized elderly patients may have dysphagic symptoms. The consequences of this disorder are significant, and aspiration pneumonia is often the outcome. Current diagnostic procedures available to identify dysphagia are discussed. PMID- 9866512 TI - Decreasing mental frailty in at-risk elders. AB - Frailty may be used to describe those older adults who are in precarious balance between their abilities to maintain health and function and their deficits that threaten the balance. Frailty often is described in purely physical terms, omitting the mental and psychologic aspects. When the term is applied to the mental abilities of older adults, frailty is associated with memory vulnerability in the cognitive domain and depression in the psychologic domain. Compromised thinking, anxiety, and decreased confidence in memory are symptoms of mental frailty. This article examines the effects of aging on memory and the intertwining factors of depression and self-efficacy as treatable antecedents of mental frailty in older adults. The article also describes simple assessment techniques, explores memory improvement strategies, and considers nursing implications. PMID- 9866513 TI - The Heritage Harbour Health Group: doing it our way. AB - The Heritage Harbour Health Group is a privately run, nonprofit health care organization founded by the residents of a retirement community for their own use. Such a facility is an original concept, and this article describes how home health care is being provided with a unique blend of professional and volunteer cooperation. Health care professionals may find information and inspiration to conduct their own research into similar programs. PMID- 9866515 TI - Nurse and judicial advocate for the frail aged: Mary Joy Quinn, RN, MA.. Interview by Priscilla R. Ebersole. PMID- 9866514 TI - Preceptors in long-term care. 1997 NGNA innovations in practice award winner. AB - A preceptor is an experienced and competent staff member who serves as a clinical role model and resource person to newly employed nursing personnel. Although this concept has been prominent since the late 1970s in acute health care settings, preceptors rarely are used in long-term care facilities. This article describes the overall effectiveness of the first long-term care preceptor program implemented at the Masonic Home of New Jersey in 1989. Conclusions will support the program as viable and beneficial to long-term care facilities by decreasing nursing turnover, providing continuity in the orientation process, promoting job satisfaction, and providing quality resident care. PMID- 9866516 TI - Frail elders: handle with care when using medications. PMID- 9866517 TI - Home visit safety. PMID- 9866518 TI - Dysphagia diet update. PMID- 9866519 TI - President's message: chaos as a positive force. PMID- 9866520 TI - Like caterpillar to butterfly: change is also a metamorphosis. PMID- 9866521 TI - Role of ophthalmic nurses with visually impaired patients. AB - In 1987, it was estimated that over 13 million people in the world were visually impaired, and more than 27 to 35 million were classified as blind. Greater than 6 million visually impaired people live in the United States. There are many causes of visual impairment, including age-related macular degeneration, cataracts, glaucoma, diabetic retinopathy, autoimmune deficiency syndrome, and trauma. Some patients may be helped or visual loss delayed with surgery. Many may have dramatic visual improvement with optical devices, large-print publications, and improved lighting. The ophthalmic nurse may assist in accurately assessing the patient's vision and identifying his or her lifestyle and ability to function independently. Assisting the patient to overcome psychosocial and economic barriers and helping to obtain assistance is critical to enable patients to function independently. Ophthalmic nurses may be instrumental in increasing patients' awareness of community resources and services available to those who are visually impaired. PMID- 9866522 TI - Tonometry. AB - Tonometry is the measure of intraocular pressure. There are several methods used to obtain intraocular pressure, such as Goldmann applanation tonometry, Tono-Pen tonometry, pneumotonometry, and tactile tension. All these methods have advantages and disadvantages. Measurement of intraocular pressure plays an important role in the detection and management of glaucomatous conditions. It is important that the examiner be aware of the various methods and their shortcomings to obtain the most accurate measurement. By obtaining the most accurate intraocular pressure by tonometry, the diagnosis and management of glaucoma can be enhanced. PMID- 9866523 TI - Role of the nurse reviewer in the preoperative setting in a specialty hospital. AB - As today's health care professionals, we are constantly challenged to provide high quality, yet cost-effective, care. Long-forgotten are the days of preadmission and extended length of stay. Health care providers are hard-pressed to accomplish much more in less time. Our hospital has taken this challenge seriously. One of the many steps that has been taken is the development of a preadmission center. The focus of this article is to describe the role of the nurse reviewer in this center, emphasizing how this pivotal role has increased efficiency, enhanced staff relations, and promoted patient satisfaction. PMID- 9866524 TI - Writing tips for authors. AB - Writing an article for publication may seem intimidating to you at first, but believe it or not, it can be exciting and fun. When you initially accept a writing assignment, research a clinical "gut feeling," or describe a nursing scenario, it may seem like a gargantuan task, but when you break it into small segments or steps you may be surprised how quickly you will complete your project. This article will offer some helpful hints or tips to get you started. Many of the tips will apply specifically for submission of an article to insight; absorb all clues that might be helpful and disregard anything that won't help you. Remember, you can get your article published, and the insight editorial board and peer reviewers only want to facilitate your success! PMID- 9866525 TI - Systemic disease and ocular disorders. PMID- 9866526 TI - President's message: working together to promote excellence in geriatric care. PMID- 9866527 TI - Patient education for decision making: more important now than ever! PMID- 9866528 TI - Operating room time is a terrible thing to waste: an operating room work improvement project. PMID- 9866529 TI - Driving with a visual impairment. AB - Driving is a highly prized activity for individuals of all ages. The decision to continue driving after a decrease in central acuity or visual field can be a source of anxiety for the individual and his or her family, as well as the ophthalmic practitioner. Although a great deal of research has been done concerning vision and driving, much controversy still surrounds this subject. This article reviews the literature concerning visual impairment and driving and makes suggestions for what should be considered when determining who should and should not continue to drive. PMID- 9866530 TI - The ophthalmic registered nurse's responsibility to the adult patient with low vision. AB - A key goal in the care of adult patients with low vision is to help them focus on their visual abilities rather than their disabilities. This article presents information that every ophthalmic nurse can use to teach patients with low vision some practical ways to change their personal environment to enhance visibility and safety. There are also some myths and misunderstandings about low vision that the nurse can dispel. Before referring patients to low-vision specialty clinics or organizations, every ophthalmic nurse can begin the visual rehabilitation for patients with low vision. PMID- 9866531 TI - Ocular injuries from golf and racket sports. PMID- 9866532 TI - Frequently asked questions about certification. PMID- 9866533 TI - Retinal detachment. AB - A retinal detachment is often traumatic and shocking to patients once they become aware of the condition. Nurses have the unique opportunity to assist these patients in accepting and in coping with this potential serious vision loss through understanding, education, rehabilitation, and ocular safety. PMID- 9866534 TI - A complication of vascular access device insertion. A case study and review of subsequent legal action. AB - A patient experienced a complication of vascular access device insertion, and his family brought charges of malpractice against the nurses and physicians involved in his care. Nurses can reduce the likelihood of being named in a lawsuit by maintaining clinical competency; communicating effectively; adhering to policies, procedures, and standards of care; and thoroughly documenting care. PMID- 9866535 TI - Technology assessment of two needleless systems. AB - A comprehensive technology assessment is used to compare two needleless systems. This technology assessment uses a structured framework for comparison of the Interlink Needleless System and the Clave, a one-piece i.v. needleless connector. A literature review and statistical data analysis are included. PMID- 9866536 TI - The reliability of blood sampling from peripheral intravenous infusion lines. Complete blood cell counts, electrolyte panels, and survey panels. AB - To assess the accuracy of venous blood sampling obtained from peripheral intravenous infusion lines, peripheral saline locks, and by peripheral venipuncture, dual blood samples from 55 hospitalized patients were compared for test reliability. Tests included complete blood cell counts, electrolyte studies, and general survey panels. Results showed that within certain limitations, blood sampling from a wide range of peripheral indwelling devices is reliable for many routine blood tests. PMID- 9866537 TI - Accidental occupational exposure of intravenous nurses to human immunodeficiency virus. Anticipating the consequences. AB - This descriptive study randomly surveyed all 302 Massachusetts members of the Intravenous Nurses Society in 1991 regarding their perceptions of nine possible consequences of human immunodeficiency virus (HIV) infection caused by accidental occupational exposure. Areas of highest concern were financial: adequacy of worker's compensation, ability of the employer to cover all healthcare costs, and job security. Nurses also were concerned about confidentiality of their HIV status and personal history jeopardizing their benefits. The i.v. nurses felt most secure in areas of their personal lives: housing and support of family and friends. Although some concerns correlated significantly with fear of contagion, others were unrelated, indicating a need for policy and attitude changes to promote comfort in working with HIV. PMID- 9866538 TI - Alternate routes of administration. AB - The less common methods of medication administration that used to be used only in the hospital are also now used in alternate settings. Understanding the treatment modalities, care requirements, and complications of these alternate routes of administration is important for nurses who participate in the care of patients receiving infusions. Several alternate routes of administration are discussed, with emphasis on the more commonly used methods: intra-arterial, intraspinal, intraventricular, intraperitoneal, intraosseous, and several other routes. PMID- 9866539 TI - Developing extravasation protocols and monitoring outcomes. AB - Vesicant extravasation is a rare adverse reaction that occurs with vesicant chemotherapeutic agents. The result of an extravasation is distressing to the nurse and patient because significant pain, tissue necrosis, and loss of function at the site of injury can result. Although numerous studies have been conducted on vesicant management, controversies still exist concerning the optimal management procedures. However, it is clear that the most effective key to management of extravasation is prevention. Standards of practice provide the basis for preventing an extravasation, with the outcome being no occurrence of extravasation. PMID- 9866540 TI - Message from the chief executive officer. PMID- 9866541 TI - Healthcare reform: its impact on transplantation. PMID- 9866542 TI - Direct contact between donor families and recipients: crisis or consolation? AB - A dramatic growth has been seen in the number of organ procurement organizations, transplant programs, and new initiatives related to solving the organ shortage, as well as in the development of protocols dealing with the logistics of facilitating organ and tissue recovery. Extensive deliberation on critical issues related to the organ donor shortage is evident in professional meetings, publications, and educational endeavors. Little research has been conducted, however, regarding direct contact between donor families and recipients, a highly controversial subject in transplantation and procurement. Should there be contact between donor families and recipients? What format should such contacts take? Who should decide? These are questions that must be considered in the field. This article will explore an organ procurement organization's 5-year experience regarding direct contact between donor families and recipients. PMID- 9866543 TI - Texas non-donor-hospital project: a program to increase organ donation in community and rural hospitals. AB - Identifying and recovering donors from community and rural hospitals present a challenge to organ procurement organizations. A study of non-donor hospitals in the United States was undertaken at Johns Hopkins University, which identified 31 hospitals (in one service area) with the facilities to accommodate organ donation, though an organ donor had not been produced in 3 years. The purpose of this study was to determine whether donors could be produced from these hospitals. A large, geographically dispersed OPO initiated a program consisting of (1) in-house coordinators, and (2) routine notification of all hospital deaths. Following implementation of this program, organ donation increased 387% among the targeted 25 hospitals. The number of hospitals producing at least 1 organ donor increased 133%. The number of organs recovered in the project increased 449%. In-house coordinators, by identifying potential donors and facilitating an organ donor awareness program, can increase the number of organ donors in hospitals with low, but real, donor potential. PMID- 9866544 TI - Attitudes and beliefs concerning organ donation among Native Americans in the upper Midwest. AB - Because of increasing access to renal replacement therapy and the high incidence of renal disease among Native Americans largely due to type 2 diabetes mellitus, it is important to determine attitudes and beliefs in this population toward organ donation. In a study conducted on reservations in the upper Midwest during traditional powwows and health fairs, it was discovered that willingness to donate was more likely to occur if Native Americans were approached by a healthcare worker from their culture, if they had already signed a donor card, or if they knew someone with diabetes. Willingness to be a living donor (81%) was greater than willingness to donate after death (i.e., as a cadaveric donor) (54%). These findings indicate that cultural-specific information about organ failure rates and organ donation, when presented by knowledgeable individuals within the culture, could increase donation in the Native American population. PMID- 9866545 TI - Islamic views on organ donation. AB - As medical technology continues its advance, the use of organ transplantation for the palliation and cure of chronic diseases is rising. However, many barriers to organ donation exist, including religious ones. It has long been known that Muslims in North America tend not to donate organs. In the past this tendency has been attributed to religious prohibitions even though cultural views may also play a strong role. The purpose of this article is to explore and define the Islamic religious opinion on organ donation and transplantation to enhance healthcare professionals' understanding of Islamic views concerning brain death and organ donation. It is hoped that this knowledge and understanding may benefit both patients and caregivers in the North American healthcare setting. PMID- 9866546 TI - Obesity and hypercholesterolemia following heart transplantation. AB - Successful strategies for avoiding obesity and hypercholesterolemia are difficult to validate because of imprecise problem identification. The purpose of this study was to describe the incidence, severity, and onset of obesity and hypercholesterolemia among heart transplant recipients and identify relationships between demographic variables and weight or cholesterol levels during the first year following transplantation. Data were collected from retrospective chart review. Forty-two patients were randomly selected from 224 patients who were undergoing heart transplantation at the Johns Hopkins Hospital between July 1983 and December 1995. Significant differences were found in weight and cholesterol level during the first 12 months. Patients with ideal body weight less than 110%, compared with greater than 110%, survived longer. Relationships were identified between prednisone dose and weight, cumulative prednisone dose and weight, and weight change and change in total cholesterol level 1 year following transplantation. Multivariate analysis showed cumulative prednisone as an independent predictor of weight. Obesity and hypercholesterolemia were significant problems within 3 months of transplantation. Although prednisone dosage should be adjusted to the lowest possible dose, dietary and lifestyle changes remain the foundation of effective management of these posttransplant complications. PMID- 9866547 TI - Family caregivers' burden, quality of life, and health following patients' renal transplantation. AB - Improved quality of life and physical functioning among renal transplant recipients have been documented; however, little of the literature has addressed the effects of transplantation on family caregivers. The purpose of this exploratory descriptive study was to characterize the level of subjective burden, quality of life, and self-rated health of caregivers who assist family members prior to transplantation as well as at 6 months following. The study sample included 19 caregivers of 19 renal transplant recipients. In general, caregiver burden, quality of life, and self-rated health did not improve following patients' transplants. In addition, 9 of 19 family caregivers reported increased burden. Because much of healthcare is family-based and greater reliance on family support seems inevitable, further studies are needed to examine the impact of transplantation on the family as well as the impact of the family on patients' posttransplant outcomes. PMID- 9866548 TI - Perceived stressors and coping strategies among heart transplant candidates. AB - This study examined heart transplant candidates' perceived stressors and coping strategies at the time of placement on the waiting list and at 3, 6, 9, and 12 months following. Mean stress scores were relatively low at each assessment time. The 4 greatest stressors were (1) having a terminal disease, (2) needing a heart transplant, (3) worrying family members, and (4) undergoing prolonged hospitalization. Total stress scores indicated that candidates were effectively using a moderate number of coping strategies, the 5 most frequently used ones being thinking positively, trying to keep life normal, keeping a sense of humor, praying or trusting in God, and trying to distract oneself. The 5 most effective coping mechanisms were thinking positively, keeping a sense of humor, thinking of good things, praying or trusting in God, and trying to keep life normal. No significant differences were seen in coping use or effectiveness over time. PMID- 9866549 TI - Analysis of organ donation in Italy. AB - In 1995 the Italian Institute of Health began assembling information regarding organ donors, recipients, and transplants. Data on more than 3000 transplants were collected and recorded to date. In addition to characteristics of donors and donating hospitals, this article describes donation activity and organ procurement in Italy from 1992 to 1996. Despite the encouraging results, the donation situation in Italy remains a challenge due to the nonhomogeneous distribution of organ donation throughout the country. It is hoped that an increase in the rate of organ donation and procurement in some regions will enable Italy to reach the European mean. PMID- 9866550 TI - Nursing faculty: positions, recruitment and retention in NLN associate degree schools of nursing. PMID- 9866551 TI - A hands-on activity for the review of the renin-angiotensin-aldosterone system. PMID- 9866552 TI - Turning a stumbling-block into a stepping-stone. PMID- 9866553 TI - The relationship between reduced clinical hours and critical thinking skills. PMID- 9866554 TI - Promoting student questioning. AB - Although critical thinking is an essential student skill, strategies that promote critical thinking are not always employed in nursing education. Fostering student questioning is one strategy that can be integrated in both the classroom and clinical setting. Some research results indicate that student questioning improves problem solving. However, students told to seek answers must first learn to ask questions. Students can be taught and encouraged to question through a variety of methods the author discusses in this article. PMID- 9866555 TI - A chemical dependence clinical experience. AB - Chemical dependence treatment clinical experiences in undergraduate nursing programs can provide appropriate entry-level experiences for students in community settings when supervised adequately and arranged carefully. They increase students' awareness of the pervasiveness of alcohol and substance related problems and its impact on individuals, families, and societies. With careful attention to entry barriers and orientation issues, faculty can develop clinical sites that provide a wealth of experience and opportunity for students to develop themselves both personally and professionally. PMID- 9866556 TI - A course in nursing consultation. Promoting indirect nursing activities. AB - Skills in consultation are a core competency for nurses practicing in advanced roles. Although changes in the healthcare delivery system are creating a growing need and increasing opportunities for nurses to provide consultation services, nurse educators have been slow to respond to the need for students to have formal education in consultation concepts and processes. The author describes a graduate level course in nursing consultation. Suggestions are given for incorporating consultation content into other courses at both the graduate and undergraduate level. PMID- 9866557 TI - Student nurses' attitudes toward nursing research. AB - This study has various limitations and some generalizations have been made in the interpretations of the findings. However for nursing research to be of significance to students as a means of inquiry, problem solving and investigation, research concepts need to be introduced in nursing education programs as early as possible. Emphasis could be placed on demonstrating processes, application, participation, and environment. In conclusion, encouraging participation, a more comprehensive connection between research processes and demonstrating the relevance of research to nursing is crucial to students in preparing them for their impending role as a registered nurse. PMID- 9866558 TI - Student projects enhance competency in community-based nursing practice. AB - Trends in healthcare delivery toward community-based care challenge nurse educators to design nursing curricula and learning experiences that prepare students for practice in community settings. The authors describe how faculty used student projects to enhance community learning experiences. The projects were designed to help students gain skill in the process of program design and evaluation and an appreciation and enthusiasm for community-based nursing practice. PMID- 9866559 TI - Nursing informatics needs assessment. Are distance programs needed? AB - Nursing informatics is a small but growing specialty area in nursing. The authors describe the results of a needs assessment designed to determine interest in a distance-based master's degree and certificate program in nursing informatics. Information was also collected on computer and Internet proficiency for a large sample of BSN graduates. Results suggest that there is an interest in informatics, although somewhat less than was found previously. Respondents indicated that a knowledge of nursing informatics could provide additional career opportunities and that there was a general lack of programs available. PMID- 9866560 TI - International education. A United Kingdom nursing student partnership. AB - An international educational exchange between nursing students in the United States and the United Kingdom provides the critical link to develop global partnership to share and exchange information and knowledge on nursing and healthcare systems. This study program assists nurses to view themselves as part of the global community and encourages cooperation in advancing the vision of healthcare reform worldwide. PMID- 9866561 TI - Teaching introductory statistics to baccalaureate nursing students. PMID- 9866562 TI - Concept mapping. Navigating the learning process. AB - Critical thinking, meaningful learning, and concept synthesis are key phrases pervading nursing literature as educators strive to promote optimal learning and improve clinical instruction and problem solving across levels of nursing education. Concept mapping offers a state-of-the-art metacognitive approach to achievement of these important goals. The author explores concept mapping from the perspective of cognitive theory, analyzes its strengths and drawbacks, presents examples of various concept maps and graphic organizers, and suggests pragmatic applications to nursing clinical, classroom, curriculum, and research situations. PMID- 9866563 TI - The nursing process as a model for clinical supervision. PMID- 9866564 TI - Peer review through clinical rounds. A collaborative critical thinking strategy. AB - Facilitating the development of critical thinking in beginning nursing students is challenging and exciting. Peer review through student-led clinical rounds can be an effective way to promote students' critical thinking in communicating assessment data and identifying client strengths and problems. Nineteen beginning baccalaureate nursing students participated in a peer review clinical round experience and were then asked to describe their critical thinking. Strengths of using the peer-led rounds include promotion of active student learning and encouragement of collaboration among peers. PMID- 9866565 TI - Searching for the dean. Contemporary and traditional considerations. AB - The process of searching for a new dean is a challenge. Access to new technologies and increasing demands placed on nursing faculty and administrators may alter traditional search strategies as well as committee composition and technical support. The author addresses the impact of these changes on the dean search process. In addition, the author details the essential tasks of a search committee. PMID- 9866566 TI - New European Union directives. PMID- 9866567 TI - Economic fallout. PMID- 9866568 TI - A hard return for your money. PMID- 9866569 TI - Tricks of the trade. PMID- 9866570 TI - They're cheap, but not cheerful. PMID- 9866572 TI - Back breaking work. PMID- 9866573 TI - The price of hands-off care. PMID- 9866571 TI - World Mental Health Day. PMID- 9866574 TI - To lift or to leave? PMID- 9866575 TI - Storm warning. PMID- 9866576 TI - Lawful conduct. PMID- 9866577 TI - Inner strength. PMID- 9866578 TI - Eye care--4. Irrigation of the eye. PMID- 9866579 TI - Face to face. Interview by Eileen Fursland. PMID- 9866580 TI - A lack of attention to infection control procedures. PMID- 9866581 TI - Using teamwork to improve communication. PMID- 9866582 TI - An exploration of the value of specialist neurosurgical nurses. AB - The differences in the knowledge and skills of 75 nurses working in a variety of practice settings were examined using a case study vignette of a neurosurgical patient developed by researchers. The case study tested nursing care throughout a patient's hospital experience, including admission, presurgery, postsurgery and discharge preparation. The findings showed that nurses who had specialised in neurosurgery scored markedly higher, in all aspects of practice knowledge, than expert nurses in other specialties and general nurses. Patients undergoing neurosurgery are safer if they have nurses who are experienced, skilled and knowledgeable to care for them. PMID- 9866583 TI - The nurse's role in the care of people with MS. PMID- 9866584 TI - Preoperative fasting. PMID- 9866585 TI - Assessing the capacity to give consent. PMID- 9866586 TI - Family planning for women with learning disabilities. PMID- 9866587 TI - Publicity coup. PMID- 9866588 TI - Asking for trouble. PMID- 9866589 TI - The nature of nurture. PMID- 9866591 TI - Qualified to patronise. PMID- 9866592 TI - A case for non-experimental studies. PMID- 9866590 TI - Self-knowledge. PMID- 9866593 TI - Hustle while you wait. PMID- 9866594 TI - The ethics of teamwork. AB - Some tasks are better accomplished by a successful team effort. Teams do not replace individual achievement; however, some ethical dilemmas cannot be resolved by individuals. Teams and teamwork are defined within the context of a case study. Types of teams, factors that strengthen teams, and circumstances that threaten teams are presented. PMID- 9866595 TI - Construct validity of Minimum Data Set items within the context of the Braden Conceptual Schema. AB - The Minimum Data Set Plus (MDS+) is used by all federally-funded long-term care facilities to assess residents and plan their care. Eighteen conditions, known as Resident Assessment Protocols (RAPs) including a pressure ulcer RAP, require a written care plan by staff. Consequently, it is important to determine if the operational definitions from the MDS+ are consistent with theoretical constructs from a conceptual schema representative of pressure ulcer risk. Examination of construct validity shows that not all of the variables in the Braden Conceptual Schema are adequately operationalized by the MDS+ or the pressure ulcer RAP. Physiologic markers of nutritional status and friction/shear are not included in the MDS+. Sensory perception is included in the MDS+ but not the pressure ulcer RAP. If the MDS+ pressure ulcer RAP is to be used to plan care and assess risk, then all items should be included. The purpose of this article is to examine the construct validity between the operational definitions of risk factors from the MDS+ and pressure ulcer RAP with theoretical constructs from the Braden Conceptual Schema. PMID- 9866596 TI - The effects of ultraviolet radiation on antibiotic-resistant bacteria in vitro. AB - Wound infections produced by antibiotic-resistant bacterial strains are particularly difficult to manage. This study examined the effectiveness of ultraviolet (UV) light treatment in killing antibiotic-resistant strains of Staphylococcus aureus and Enterococcus faecalis in vitro. Between 2 and 5 replications of each organism at 10(8) organisms/ml were prepared and plated on sheep blood agar medium and treated with UV light (254 nm, 15.54 mW/cm2 output). Irradiation times were 0, 2, 5, 8, 15, 30, 45, 60, 90 or 120 seconds. Bacterial cultures were then incubated at 35 degrees C for 24 hours. Kill rates were 99.9 percent for the methicillin-resistant strain of S. aureus (MRSA) at 5, 8, 15, 30, 45, 60 seconds and 100 percent at 90 and 120 seconds. Kill rates were 99.9 percent at 5, 8, 15, 30 seconds for vancomycin-resistant E. faecalis (VRE) and 100 percent at 45, 60, 90, 120 seconds. Similar results were found with UV light treatment of the antibiotic-susceptible strains of S. aureus and E. faecalis. A significant difference in kill rates at 30 seconds of UV exposure was detected between the antibiotic-resistant strain of S. aureus and the antibiotic-resistant strain of E. faecalis (Student's t test, p < 0.01). Significant differences were also detected in the kill rates at 30 second exposure times for the antibiotic susceptible strains of S. aureus and E. faecalis. These findings suggest that the Enterococcal bacteria is more susceptible to the killing effects of UV. This data also suggests that UV light at 254 nm is bactericidal for antibiotic-resistant strains of S. aureus and E. faecalis at times as short as 5 seconds and that the enterococcal bacteria is more susceptible to the killing effects of UV. With recommended patient treatment times for infected wounds being significantly longer than 5 seconds, this data indicates that patient treatment times need to be re-examined. PMID- 9866597 TI - Closure of the "non-healing wound" corresponds with correction of weight loss using the anabolic agent oxandrolone. AB - We studied the relationship between restoration of weight loss and healing of the "non-healing wound." Eight consecutive patients with large "non-healing wounds" of an average of 12 months duration, despite good local wound care, were studied. All had a weight loss of 10 percent or more of body weight, mainly lean body mass. Optimizing nutrition (4 weeks) did not significantly increase weight or healing. The addition of the oral anabolic agent oxandrolone increased restoration of weight (4 lbs/week) over a 12-week period. Five wounds completely closed and three closed by 75 percent during this period. We noted a significant correlation (r = 0.67) between restoration of weight loss and closure of the previously "non-healing wound." The rate of wound healing was most prominent after 50 percent of weight loss had been restored. This finding reflects the key relationship between restoring body weight, body protein stores, and wound healing. PMID- 9866598 TI - Is intermittent pulsatile pressure a valuable adjunct in healing the complicated diabetic wound? AB - Diabetic neuropathic ulceration and subsequent lower extremity amputation are a significant cause of chronic disability. High-risk diabetic patients with infected foot wounds, poor lower extremity blood flow, and inadequately controlled blood glucose require prompt action by clinicians in order to successfully avoid more severe sequelae. Dysvascular patients who are poor candidates for arterial bypass surgery may benefit from adjunctive treatment with a pneumatic pedal compression device. The authors discuss the rationale behind this unique treatment modality and present a case report illustrating a successful outcome in a high-risk diabetic patient. PMID- 9866600 TI - Believe it or net. PMID- 9866601 TI - A revolution called managed care. PMID- 9866602 TI - Ethical dilemmas in pain management. AB - Pain management is becoming an increasingly important component of comprehensive wound care. Choices for the best approach in managing the elderly patient with painful wounds can pose ethical dilemmas. Ethical principles such as autonomy, beneficence, and non-maleficence are explored through a case presentation. Suggestions for addressing these ethical dilemmas are examined. PMID- 9866603 TI - An approach to leg and foot ulcers: a brief overview. AB - Legs and feet are susceptible to ulcer formation. Three main types of lower extremity ulcers are venous, arterial, and neurotropic. Ulcer care should include treatment of the underlying cause, moist interactive healing, and quality of life (pain control). If the ulcer shows no signs of healing in 6 to 12 weeks, the wound should be biopsied, the diagnosis confirmed, and a plan of care instituted. In certain instances, biologicals may be used. A graphical representation of an approach to ulcer care, in addition to common differential diagnoses of leg ulcers, is presented. PMID- 9866604 TI - Painful venous ulcers: themes and stories about their impact on quality of life. AB - The purpose of this dissertation research was to describe, to understand and to interpret the meaning of the experience of living with painful venous ulcers. Heideggerian hermeneutic phenomenology, a descriptive, phenomenological approach, was the design used for this study. Fourteen people with active painful venous ulcers at the time of initial interview were engaged in conversation. The semi structured interviews were audiotaped, transcribed and analyzed using Martin Qualitative Analysis Software (version 2.0). Four of the eight most compelling themes that relate to quality of life are reviewed here: feeling frustrated; interfering with the job; making significant life changes; and finding satisfaction in new activities. Increased sensitivity to and understanding of the impact of painful venous ulcers on QOL may lead to more effective intervention strategies and improved outcomes for these patients. PMID- 9866605 TI - Venous leg ulcers. AB - Venous ulcers are the most common form of leg ulcers and their incidence is increasing as the population ages. The diagnosis should be confirmed with tests for venous insufficiency and an assessment of arterial disease determined by a Doppler Ankle-Brachial ratio. The most important component of venous ulcer treatment is the use of high compression bandages for pitting edema. Non-healing ulcers may require a skin biopsy of the edge to confirm the diagnosis, medical treatment for woody fibrosis, adjustment of compression therapy, or use of a skin substitute or biological preparation. PMID- 9866606 TI - Epidemiology, risk factors, and management of peripheral vascular disease. AB - Peripheral vascular occlusive disease is initiated with a genetic risk factor component compounded by patient-controlled contributions including obesity, diabetes, hypertension, and smoking. Medical management of these factors may delay or obviate surgical intervention. Angiography may be used to perform angioplasty (+/- stents) or to guide various interventional procedures. The major contribution to pre- and post-operative assessment is the noninvasive laboratory. PMID- 9866607 TI - Diabetic foot ulcerations: an overview. AB - The development of a diabetic neuropathic ulcer is a multifactorial process with an emphasis on mechanical abnormalities. In the absence of vascular compromise, up to 90 percent of neuropathic ulcers will heal with proper ulcer debridement, treatment of infection and relief of weight from the ulcerated area. A thorough understanding of the etiology of a neuropathic ulcer is an essential component in the appropriate management of these patients. Treatment of the diabetic neuropathic ulcer is a multidisciplinary clinical problem and each member of the team has a unique role to play. PMID- 9866608 TI - Latex allergy. An institutional approach, questionnaire and information for healthcare workers. PMID- 9866609 TI - Continence care. PMID- 9866610 TI - Sex education for nurses. PMID- 9866611 TI - Practising at a higher level. AB - New nursing roles are continually being developed. Nurses working at higher levels of practice are only partially regulated at present. More formal regulation by the UKCC would help protect nurses working at higher levels, as well as their employers and patients. PMID- 9866612 TI - Exploring sexuality and sexual health in nursing. AB - In order to provide acceptable care, nurses need appropriate knowledge, attitudes and skills. Nurse education in relation to sexuality has tended to focus on the knowledge and attitude components of sexuality, but a skills component must also be included. Experiential learning activities should be explored by nurses and nurse educators as a means of developing effective and sensitive ways of working with patients' sexuality and sexual health-care issues. PMID- 9866613 TI - Setting up a nurse-led anticoagulant clinic. AB - People on anticoagulation therapy need regular monitoring. With appropriate training, nurse specialists can improve patient care in anticoagulant clinics. PMID- 9866614 TI - The value of systematic reviews in research. AB - Nurses may be deterred from using research to inform their practice because of the huge numbers of papers available on some topics. Literature reviews on a topic of interest may make the task easier. A systematic review aims to discover all the research undertaken on a subject and review all the results in a scientific manner. Systematic reviews may highlight findings not available from individual studies and can thus help to inform and improve practice. PMID- 9866615 TI - Developing a continence care programme. AB - A care programme approach to the care of patients with continence problems provides a framework within which a multidisciplinary team can work. The approach ensures patients are followed up at all stages of their care and clear referral pathways are provided. PMID- 9866616 TI - Children's cognitive level and perception of pain. AB - Doctors and nurses tend not to approach children according to their developmental level. Children develop an understanding of illness and of the cause and effect of pain that reflects Piaget's stages of cognitive development. Until nurses begin routinely to consider a child's developmental level when assessing pain, children will continue to suffer unnecessary pain and anxiety. PMID- 9866617 TI - Nebuliser therapy. PMID- 9866618 TI - The use of enteral and parenteral feeding. AB - Nutritional support involves a number of disciplines; collaborative decision making by a nutrition support team is recommended. Unless factors exist which preclude enteral feeding or make the exclusion of nutrients from the gut appropriate, every effort should be made to feed enterally. Nutritional support involves the provision of safe equipment, treatment protocols and procedures, a training programme for all staff involved in the patient's care, and an audit system. PMID- 9866619 TI - Compression hosiery. AB - Compression hosiery is used in the management of venous incompetence, which may result in varicose veins, venous ulceration, deep vein thrombosis and lymphoedema. Accurate patient assessment and measurement is essential to ensure that hosiery is appropriate, fits properly and is worn by the patient. PMID- 9866620 TI - Bereavement. Part 1: Theories of bereavement. AB - Bereavement is often accompanied by a deterioration in physical and emotional well-being. Nurses often find it difficult to deal with bereaved people, and avoid involvement with them. Understanding the bereavement process can help nurses to provide effective care and support for bereaved people. PMID- 9866621 TI - Peri-operative nursing. PMID- 9866622 TI - Latex allergy update. PMID- 9866623 TI - Latex hypersensitivity: the IgE response. AB - The IgE-mediated allergic response involves a series of physiological pathways that result in the array of symptoms commonly associated with allergy. This article describes the mediators that prompt specific clinical responses and identifies those at risk for latex hypersensitivity. PMID- 9866624 TI - Understanding latex allergy. AB - Latex allergy is an increasingly important problem in both health-care workers and patients. Predisposing factors to development of latex allergies include a history of atopy or allergy and frequent exposure to latex products. Identified allergens include latex proteins from the rubber tree that remain in manufactured products, as well as smaller molecules that remain from the latex purification and manufacturing process. Latex proteins absorbed to powder in latex surgical and examination gloves may be aerosolized and inhaled. Powder-absorbed latex proteins are thought to be important in triggering of sensitization in susceptible individuals, as well as in elicitation of symptoms in previously sensitized patients. Allergic reactions to latex can include local dermal reactions or generalized immediate hypersensitivity (anaphylactic) reactions. Pathophysiology, signs and symptoms, and treatment of each type of reaction are discussed. Measures to address latex allergy, however, must include measures to decrease exposure to latex antigens both in latex-allergic subjects, to prevent symptoms, and in naive subjects, to prevent sensitization. These measures may include finding, acceptable substitutes for latex in many products. PMID- 9866625 TI - Latex allergy and gloving standards. AB - In response to universal precautions, latex gloves are used more frequently in health care settings, and more health care workers are reporting latex sensitivity or allergic reactions to natural rubber latex (NRL). As a result, there continues to be controversy about what gloves should be used in clinical practice and what are the appropriate clinical recommendations about gloving practices. This article reviews the current issues about gloving practices, addresses national standards related to gloving, and makes recommendations about future gloving practices. PMID- 9866626 TI - Why latex allergy now? AB - Type I allergy to latex did not appear in the literature until 1979. For over 100 years people have used natural rubber latex products without apparent problems. Why are allergies appearing now? Scientists are not absolutely sure. Reasons may include an unprecedented increase in the use of latex gloves, changes in the manufacturing process by some manufacturers, and the switch to one-time-use disposable gloves. To reduce the risk of latex protein allergy, health care workers should use nonlatex gloves for activities that are not likely to involve contact with infectious materials. Appropriate barrier protection is necessary when handling infectious materials. When using latex gloves, use powder-free gloves with reduced protein content. Sensitized individuals should use only nonlatex gloves. PMID- 9866627 TI - Identifying the risk. AB - This article will provide the perioperative nurse with some basic information for identification of latex-containing supplies and basic guidelines for creating a latex-free environment, including the setup and contents of a latex allergy cart. PMID- 9866628 TI - Are nurses knowledgeable in regards to latex allergy? AB - Lack of knowledge concerning latex allergy may lead to a life-threatening adverse reaction to natural rubber latex. Registered nurses need a latex allergy knowledge base to provide latex-safe health care to clients and to create a latex safe environment. This research explored the current knowledge base of registered nurses as related to (1) the care of clients at risk for latex allergy, or diagnosed with latex allergy, and (2) the provision of a latex-safe environment for all populations. This article describes this research and its implications for nurses. A copy of the Latex Allergy Knowledge Base Self-Assessment Questionnaire is included at the conclusion of the article. PMID- 9866629 TI - Living and working with latex allergies: personal perspectives from a nurse. AB - Living with the limitations of latex sensitivity can be devastating. Even more devastating is when the diagnosis can end a career. For a registered nurse, latex allergy can be a career ender, or it can spur the individual to become increasingly clever in coping with the allergy while maintaining a productive personal and professional life. This article describes the personal struggle an individual registered nurse has overcome to maintain her position in nursing. PMID- 9866630 TI - Workforce forecast: hi-tech equipment, GenX employees, and guaranteed change ahead. PMID- 9866631 TI - The design of today's operating rooms. AB - With the exception of radiation therapy, the surgical suite is still the most costly area in a health care facility to renovate or construct. It is also the most needs intensive, as far as staff, equipment, material, and building system. Health care providers need an environmentally sensitive area that meets the needs of today, as well as the future. It is equally important to create a working environment that promotes staff comfort, while streamlining time and cost. PMID- 9866633 TI - OR nursing in the Net. AB - A mailing list is an agreement between people with similar interests who meet electronically. This perioperative listserver is unmoderated and open to nurses and other health professionals interested in the perioperative care of patients. PMID- 9866632 TI - Today's promising heart frontier: minimally invasive surgery. AB - Cardiovascular disease claims more lives annually than the next eight leading causes of death combined. The biggest heart problem in the United States is congested "plumbing," afflicting more than 13 million Americans. Open-heart surgery estimated to be a $30 billion-a-year business. PMID- 9866634 TI - Low-temperature hydrogen peroxide gas plasma--atomic age sterilization technology. AB - The combined use of hydrogen peroxide vapor and plasma sterilizes most medical instruments and material without leaving toxic residues. Patients and personnel are not in jeopardy of exposure to an inadequately aerated instrument this this sterilization method. PMID- 9866635 TI - Total artificial heart to usher in new age of cardiology. AB - A battery-powered artificial heart--designed to replace the diseased heart both physically and functionally--is completing readiness testing in preparation for human trials. The initial goal is for the artificial heart to last 5 years, at which time a donor heart can be transplanted. No immunosuppression will be needed with the total artificial heart, and the recipient's hospital stay will be much shorter, resulting in a tremendous decreases in cost. PMID- 9866636 TI - USP--promoting the public health. AB - As one of the first "national" medical organizations, the first pharmacopeia contributed to a united national public health effort. The USP was the precursor to the American Medical Association. An official USP drug monograph tells manufacturers what constitutes the acceptable identity, quality, strength, and purity of a pharmaceutical product. PMID- 9866637 TI - Making the choice: can you go 'latex free'? AB - Health care facilities have many means available to minimize the risk of latex allergy for both patients and workers. A latex-allergy task force should oversee patient and employee safety. Materials and items that contain latex must be identified, and latex-free alternatives must be found. PMID- 9866638 TI - Evaluation, treatment, and management of erectile dysfunction: an overview. AB - Erectile dysfunction affects approximately 10 to 20 million men in the United States. During the last decade there has been a significant change in the management of patients with sexual dysfunction both because of our improved understanding of erectile physiology, and also because of the development of new and effective medical therapies. PMID- 9866639 TI - Factors in predicting initial in-office therapeutic dosages of alprostadil for the treatment of organic impotence. AB - The use of intracavernous penile injections of alprostadil for treating organic impotence is well-established. Using blood pressure measurements, especially systolic BP and MABP as measured in a flat supine position, the dosage of alprostadil required to achieve satisfactory erections in men with organic impotence can be predicted. This guidance can significantly influence the outcome of therapy by reducing the number of titrated diagnostic injections and the time required for the patient to achieve satisfactory results. PMID- 9866640 TI - Implementing an ambulatory surgical unit in pediatric urology. AB - Health care professionals have been asked to do more with less and examine every possible option to cut costs but not service. To meet this challenge, a pediatric urology same-day admit/day care unit provides patients and their families with surgical and diagnostic care that has been well received by patients, families, and staff. PMID- 9866641 TI - The evolving role of submucosal injectables for treating internal sphincteric deficiency. AB - Each of the previously mentioned materials can be used in procedures performed on an outpatient basis using sedation and local anesthesia. Teflon usually requires general anesthesia. None of these materials, however, should be used in acute conditions involving cystitis, urolithiasis, or infection. Table 4 summarizes the postoperative care after a bulking agent injection. There are obvious advantages to using injectable materials for ISD, one of which is that the outpatient procedure is usually done under local anesthesia. It does not have the risks inherent in open surgical procedures like slings or artificial sphincters, and it causes minimal increase of urethral resistance to detrusor-generated micturition force. Such procedures are obviously easier and faster to perform with less cost and rapid recovery followed by a return to regular activity within 48 hours. In conclusion, injection therapy for urinary incontinence is appropriate, but the most effective substance has probably not yet been determined. PMID- 9866642 TI - Treatment of urinary incontinence among caregiver-dependent adults. AB - It is estimated that approximately 50% of residents in long-term care settings suffer from urinary incontinence. Developing and implementing a continence program in long-term care settings is one of the most challenging aspects of continence care. Careful assessment of residents will allow interventions to be targeted to those most likely to benefit. The combination of nursing measures and targeting the behavioral interventions to those most likely to benefit increases the likelihood that the program will produce clinically significant reductions in UI. PMID- 9866643 TI - Getting ready for certification: let's get focused! PMID- 9866645 TI - Abnormal T waves in a woman with cerebral hemorrhage. PMID- 9866644 TI - Aftereffects of a tainted meal. AB - A 58-year-old man presented with profuse diarrhea, abdominal distention, flatulence, weakness, and a 15-lb weight loss. The initial symptoms of nausea and vomiting had begun a week earlier, within hours of eating chicken that had not been well wrapped when purchased. Two days after symptom onset, he received intravenous rehydration therapy at another hospital. With resolution of the nausea and vomiting, he felt slightly better and went home against medical advice. He had not had fever, chills, hematochezia, melena, hematemesis, or dysuria. PMID- 9866646 TI - Bartonella infections: diverse and elusive. AB - Children with young cats, HIV-infected patients with low CD4 counts, and lice infested homeless people are among those with an increased risk. Clinical presentation varies with the infecting Bartonella species and the host. Treatment for cat-scratch disease is supportive; macrolide therapy is an appropriate choice for other Bartonella infections. PMID- 9866647 TI - Primary care of HIV infection. AB - Four cases illustrate some of the issues involved in treating HIV-infected patients in a primary care setting. Primary care physicians are hard-pressed to achieve the same results as infectious disease specialists, yet are increasingly responsible for performing the initial tests, choosing the therapeutic regimen, ensuring the patient's compliance with the regimen, and monitoring the results. PMID- 9866648 TI - Evaluation and management of insomnia. AB - Insomnia is common, but the causes are so diverse that careful evaluation is needed to ensure selection of effective treatment. Duration is the most important guide to intervention. Therapies include behavior modification, exercise, and medication. Drugs for patients who require long-term therapy should be selected to minimize side effects. Long-acting drugs should be used with caution in the elderly. PMID- 9866649 TI - When a leader is confronted with change. PMID- 9866650 TI - Heart failure 101. PMID- 9866651 TI - Striking, asymptomatic skin lesions in a 42-year-old man. PMID- 9866653 TI - Case in point. Inflammatory breast carcinoma. PMID- 9866654 TI - Calcium therapy for treating PMS. PMID- 9866652 TI - Case in point. Acute epiglottitis with subcutaneous emphysema. PMID- 9866655 TI - Minimizing trauma to the genital tract in childbirth. PMID- 9866656 TI - Tight blood pressure control in type 2 diabetes. PMID- 9866657 TI - Antihypertensive therapy in type 2 diabetes mellitus. PMID- 9866658 TI - Noninvasive testing for coronary artery disease: which is best? PMID- 9866659 TI - The best therapy for allergic rhinitis. PMID- 9866660 TI - Broad versus narrow spectrum antibiotics for sinusitis. PMID- 9866661 TI - Combined oral contraceptives versus levonorgestrel for emergency contraception. PMID- 9866662 TI - Hospitalism debate. PMID- 9866663 TI - Hospitalism debate. PMID- 9866664 TI - Antibiotics for acute bronchitis. PMID- 9866665 TI - Transient discontinuity of care. Others seeing what we have missed. PMID- 9866666 TI - Clinical jazz: harmonizing clinical experience and evidence-based medicine. PMID- 9866667 TI - Mupirocin cream is as effective as oral cephalexin in the treatment of secondarily infected wounds. AB - BACKGROUND: Topical antimicrobials have been considered for treatment of secondarily infected wounds because of the potential for reduced risk of adverse effects and greater patient convenience. We compared mupirocin cream with oral cephalexin in the treatment of wounds such as small lacerations, abrasions, or sutured wounds. METHODS: In 2 identical randomized double-blind studies, 706 patients with secondarily infected wounds (small lacerations, abrasions, or sutured wounds) received either mupirocin cream topically 3 times daily or cephalexin orally 4 times daily for 10 days. RESULTS: Clinical success at follow up was equivalent in the two groups: 95.1% and 95.3% in the mupirocin cream and the cephalexin groups, respectively (95% confidence interval [CI], -4.0% to 3.6%; P = .89). The intention-to-treat success rate was 83% in both groups. Bacteriologic success at follow-up was also comparable: 96.9% in the mupirocin cream and 98.9% in the cephalexin groups (95% CI, -6.0% to 2.0%; P = .22). The occurrence of adverse experiences related to study treatment was similar for the 2 groups, with fewer patients in the mupirocin cream group reporting diarrhea (1.1% vs 2.3% for cephalexin). CONCLUSIONS: Mupirocin cream applied topically 3 times daily is as effective as oral cephalexin given 4 times daily for the treatment of secondarily infected wounds and was well tolerated. PMID- 9866668 TI - Twenty-year trends in the Ohio generalist physician workforce. AB - BACKGROUND: Many factors contribute to the variations seen in physician workforce projections, including assumptions about attrition, new physician entry, and geographic requirements. Our study offers data for bench-marking future research into this complex issue. METHOD: At 5-year intervals starting in 1975, data were collected for each Ohio county by local physician census takers. RESULTS: Total Ohio family physician rates per population did not increase appreciably during the 20-year period. A decrease in the number of allopathic family physicians was balanced by an increase in the number of osteopathic family physicians, many of whom were graduates of the state's first osteopathic medical school, which graduated its first class in 1980. Rates of general internists and general pediatricians increased. In 1975, the percentage of physicians older than 59 years was higher for family physicians than for general internists and general pediatricians. By 1995, this disparity in age distribution had greatly decreased. Rural family physician rates per 100,000 population decreased, and urban rates increased, while both urban and rural rates increased for general internists and general pediatricians. CONCLUSIONS: Variations in accounting for clinical time used for non-generalist clinical and nonclinical activities may explain a large part of the difference between generalist head count and full-time equivalency (FTE) study results; together these activities can be said to make up a "fourth compartment" contributing to improper specialty designation. The decrease in the percentage of family physicians older than 59 years indicates that the future supply of practicing family physicians is not in jeopardy. The rural family physician workforce is decreasing, while the general internist and general pediatrician rural workforce is increasing, but the total rural workforce is still well below the urban workforce. Neither component of the rural workforce appears to have stabilized. PMID- 9866669 TI - Integrating the family into routine patient care: a qualitative study. AB - BACKGROUND: The field of family medicine has been enriched by a family-oriented approach and the inclusion of family systems concepts. Keeping the family as a central focus of care has been a fundamental commitment of family medicine. This research examines how exemplary physicians ("exemplars") integrate a family oriented approach into the routine care of individual patients. METHODS: Four family physician exemplars were observed. A total of 16 days was spent observing the physicians; 137 physician-patient encounters were audiotaped, transcribed, and analyzed. Grounded theory was used for analysis, and a model of a family oriented approach was developed. RESULTS: Visits were classified by the reason for visit and the intensity of family-oriented talk and actions. There was modest variation among the physicians in terms of intensity and time spent with patients. Overall, 19% of patient encounters had a high intensity of family orientedness; 34% were of low intensity. The average time spent with patients was 13 minutes, with visits ranging from 3 to 39 minutes in length. CONCLUSIONS: Our study demonstrated that physicians integrate family systems concepts into routine individual patient care. The findings identify characteristics of the family oriented approach and those circumstances that promote and hinder it. Family physicians can adapt specific components of the family-oriented approach into their routine individual patient care. PMID- 9866670 TI - Depression, health-related quality of life, and medical cost outcomes of receiving recommended levels of antidepressant treatment. AB - BACKGROUND: We evaluated depression severity, health-related quality of life (HRQL), and medical cost outcomes of primary care patients receiving recommended and less-than-recommended levels of antidepressant treatment. METHODS: We performed a secondary analysis of clinical trial data from primary care clinics in a staff-model managed care organization. The trial included patients with Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised (DSM-III-R) criteria for major depression who were starting antidepressant treatment. The primary outcomes measures used were the 17-item Hamilton Depression Rating Scale (HDRS), Hopkins Symptom Checklist depression scores, the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) mental and physical component summary scores, and the total outpatient and inpatient medical costs. RESULTS: Of 358 patients starting antidepressant treatment, 195 (54.5%) received doses recommended by the Agency for Health Care Policy and Research for 90 days or more. Mean HDRS score decreased from 14.1 to 8.8 in patients receiving less-than-recommended treatment and decreased from 13.8 to 8.9 in patients with minimum recommended treatment (P = .761). No significant differences in improvement of HRQL outcomes during 6 months were observed between patients receiving recommended or less-than-recommended antidepressant therapy. Mean total medical costs over 6 months for patients taking the recommended levels of antidepressant treatment were $1872 +/- 140 compared with $2622 +/- 413 for patients taking less-than-recommended treatment (P = .032). The differences in total medical costs were attributable to significantly lower nonmental health related inpatient costs in the recommended antidepressant treatment group ($104 vs $785, P = .004). CONCLUSIONS: Patients receiving minimum recommended levels of antidepressant therapy for 3 months showed improvement in depression severity and HRQL comparable with patients receiving less-than-recommended treatment. Patients receiving minimum recommended treatment had lower total costs and nonmental health-related inpatient costs. Antidepressant treatment in primary care patients may have the greatest impact on the frequency of health care visits and on costs for medical conditions and impairments. PMID- 9866671 TI - Are antibiotics effective treatment for acute bronchitis? A meta-analysis. AB - BACKGROUND: Most clinicians prescribe antibiotics for acute bronchitis in spite of recommendations against this practice. Because the results of individual clinical trials have been mixed, we conducted a meta-analysis to determine whether antibiotics are effective treatment for acute bronchitis. METHODS: We conducted a comprehensive search to identify all trials in which patients who had a diagnosis of acute bronchitis were randomly assigned to treatment with an antibiotic or placebo. Patient-oriented outcomes of importance that were reported in at least 3 studies were quantitatively summarized. RESULTS: Nine studies met the inclusion criteria, but not all trials provided data for each outcome. Patients given antibiotics were less likely to have a cough (relative risk [RR] = 0.69; 95% confidence interval [CI], 0.49 -0.98) and be considered unimproved (RR = 0.51; 95% CI, 0.30-0.88) at a follow-up visit; but they were not less likely to have a productive cough (RR = 0.79; 95% CI, 0.60-1.03), activity limitations (RR = 0.59; 95% CI, 0.24-1.44), or feel ill (RR = 0.70; 95% CI, 0.31-1.58). Antibiotic-treated patients had a slightly shorter duration of productive cough (weighted mean difference [WMD] = -0.56 days; 95% CI, -1.09 to -0.04), but not of overall cough (WMD = -0.94; 95% CI, -2.08 to 0.21) or activity limitations (WMD = -0.49; 95% CI, -1.07 to 0.10). Patients treated with antibiotics did not report significantly more adverse effects (RR = 1.47; 95% CI, 0.82-2.65). CONCLUSIONS: Antibiotics may be modestly effective for a minority of patients with acute bronchitis. It is not clear which patient subgroups might benefit, and the failure of some studies to report negative findings may have resulted in overestimates of the benefits of antibiotics. Antibiotics are not necessary for every patient with acute bronchitis. PMID- 9866672 TI - Evaluation and management of newborn jaundice by midwest family physicians. AB - BACKGROUND: Recognition and management of newborn jaundice is controversial and even more challenging with the early discharge of newborns. The purpose of this study was to describe the jaundice management patterns of family physicians in Minnesota and Wisconsin and compare them with American Academy of Pediatrics recommendations. METHODS: Forty-two members of the Practice-Based Research Group of the Wisconsin Research Network and of the Minnesota Academy of Family Physicians Research Network recorded information on study cards about the care they provided to healthy full-term newborns for 6 months. RESULTS: Data was collected on 335 infants, 30% of whom were jaundiced. Physicians ordered limited laboratory testing on selected jaundiced infants. Infants with jaundice were more frequently breast-fed, and had longer hospital stays. Jaundice was commonly managed by phototherapy (17%), home sunlight (28%), and increased breast-feeding (44%). Twenty-six percent of the physicians used a formal phototherapy guideline. The timing of the first follow-up visit did not differ for infants discharged before or after 48 hours of age. CONCLUSIONS: Family physicians identified and managed newborn jaundice relatively infrequently in our study. Their practice patterns were consistent with most aspects of the American Academy of Pediatrics jaundice guideline, although few of them used it to guide phototherapy use. The study physicians did not generally follow recommendations for follow-up of infants discharged early. Until a large-scale clinical trial of newborn jaundice management is performed, a variety of practices should continue to be acceptable. PMID- 9866673 TI - Decision making in everyday life. A theory-based approach. PMID- 9866674 TI - Fluorescence quenching and dequenching analysis of RNA interactions in vitro and in vivo. AB - This paper describes the use of fluorescence quenching and dequenching to analyze molecular interactions of RNA in vitro and in vivo. Fluorescein-labeled ribonucleotide was incorporated into an RNA substrate by in vitro transcription. The fluorescence quantum yield of the intact RNA was reduced by intramolecular quenching. When the RNA was degraded by ribonuclease digestion, the quantum yield increased by approximately 50%, reflecting dequenching due to separation of proximate fluorophores. Dequenching was dependent on the concentration of enzyme and substrate and was inhibited by the ribonuclease inhibitor RNasin. Comparable rates of dequenching were observed with RNase A and RNase T1. Dequenching provides a sensitive, quantitative, and convenient assay for RNA degradation. When fluorescent RNA was microinjected into cells in culture the intracellular fluorescence declined gradually with time after injection reflecting "superquenching: due to intermolecular interactions between the injected RNA and intracellular components. Capped RNA exhibited greater superquenching than uncapped RNA. Superquenching provides a sensitive, quantitative, and specific assay with subcellular resolution for intermolecular interactions of RNA in vivo. When RNase was injected into the same cells, fluorescence increased by approximately 50%, indicating that fluorescence dequenching due to RNA degradation can be measured in vivo as well as in vitro. PMID- 9866675 TI - Ligand binding to macromolecules or micelles: use of centrifugal ultrafiltration to measure low-affinity binding. AB - We describe a method for estimating ligand binding to a macromolecular sample under conditions where this binding is of low affinity and must be measured under equilibrium conditions, without removal of the unbound ligand. The method is based on centrifugal ultrafiltration through a membrane with a molecular mass cut off intermediate between that of the ligand and that of the target, and the amount of bound ligand is calculated from the difference between the (total) ligand in the concentrated sample and the (free) ligand in the ultrafiltrate. Centrifugal ultrafiltration makes it possible to separate free ligand from bound ligand (without changing its concentration) and to simultaneously concentrate the target (such that the proportion of bound ligand becomes significant, even under low-affinity binding conditions). We applied this technique, using Centricon 10 (Amicon) devices, to several cases (soluble proteins, intact membranes, detergent solubilized proteins, and pure detergent micelles) and assessed its value with respect to the common artifacts that occur in other protocols involving protein retention on nitrocellulose filters (nonspecific ligand adsorption and protein denaturation). PMID- 9866676 TI - A gel diffusion assay for quantification of pectin methylesterase activity. AB - Increased binding of ruthenium red to pectin as the number of methyl esters attached to the pectin decreases was used as the basis for a gel diffusion assay for pectin methylesterase (PME, EC 3.1.1.11) activity. The stained zone diameters resulting from the hydrolysis of 0.1% (w/v) 90% esterified pectin in an agarose gel by diffused, commercial PME were log-linear over 4 orders of magnitude, with a minimum detection limit of 3.6 pkatals. Pectin deesterification as the cause for a stained zone after PME incubation was confirmed when only 1 N NaOH, which will chemically deesterify the pectin, and not methanol or acid, the two products formed when PME acts on a methyl ester, resulted in the characteristic stained zone. The stained zone diameters decreased with increasing percentage of substrate esterification, were independent of pH, and were insensitive to simultaneous incubation with two forms of pectin lyase (EC 4.2.2.10), polygalacturonase (EC 3.2.1.15), or all combinations. PME extracted from tomato seeds, cotton fibers, and melon fruit showed pH optima of 6, 6, and 8, respectively. Using individual tomato seed parts, the assay was adapted to quantify diffusate activity and to localize activity in tissue prints. The sensitivity, specificity, and simplicity of this PME assay are superior to all others. PMID- 9866677 TI - Mapping a protein-binding site on straightened DNA by atomic force microscopy. AB - We have developed an Atomic Force Microscopy (AFM)-based method for mapping protein-binding sites on individual, long DNA molecules (> 5 kb) at nanometer resolution. The protein is clearly detected at the apex of the bent DNA molecules. Randomly coiled DNA molecules or protein:DNA complexes were extended by a motor-controlled moving meniscus on an atomically flat surface. The immobilized molecules were detected by AFM. The straightened DNA displayed a sharp bend at the site of bound protein with the two DNA segments linearly extending from the protein-binding site. Using GAL4, a yeast transcription factor, we demonstrate good agreement of the position of the observed binding site on straightened DNA templates to the predicted binding site. The technique is expected to have significant implications in elucidating DNA and protein interactions in general, and specifically, for the measurement of promoter occupancy with unlabeled regulatory proteins at the single-molecule level. PMID- 9866678 TI - An improved sample packing device for rapid freeze-trap electron paramagnetic resonance spectroscopy kinetic measurements. AB - A new method has been developed for sample packing in rapid freeze-quench electron paramagnetic resonance spectroscopy (EPR) kinetic experiments. Sample particles freeze-quenched in chilled isopentane are filtered under pressure through a stainless steel funnel attached to an EPR tube fitted with a porous disk at its bottom. Isopentane exits through the porous disk and the sample particles can be transferred essentially quantitatively into the receiving EPR tube. This device provides a more predictable, reproducible, and time-saving method for sample packing, enables use of a wider range of flow velocity, and allows efficient use of valuable reactants. PMID- 9866679 TI - Analysis of oligosaccharide structures from the reducing end terminal by combining partial acid hydrolysis and a two-dimensional sugar map. AB - A new sensitive and convenient method for the structural analysis of oligosaccharides was developed, in which partial acid hydrolysis was combined with two-dimensional sugar mapping of pyridylamino (PA)-oligosaccharides. A PA oligosaccharide was partially hydrolyzed, the acid hydrolysis conditions being optimized so as to obtain various PA-oligosaccharide fragments with high yields from different types of PA-oligosaccharides. The acid hydrolyzates were then fractionated every 1 glucose unit by size-fractionation HPLC, and each fraction was analyzed by reversed-phase HPLC. The structure of each PA-oligosaccharide fragment was identified on a two-dimensional sugar map prepared with standard PA sugar chains, after which the original PA-oligosaccharide was reconstructed from the reducing end terminal based on the obtained structures of the PA oligosaccharide fragments. The method was successfully applied to resolving the structure of an N-linked sugar chain. PMID- 9866680 TI - Measurement of plasma S-adenosylmethionine and S-adenosylhomocysteine as their fluorescent isoindoles. AB - The low levels of S-adenosylmethionine (AdoMet) and S-adenosylhomocysteine (AdoHcy) in plasma can be measured by formation of the fluorescent isoindole derivatives of these compounds. The procedure involves an initial separation of AdoMet and AdoHcy in deproteinized plasma by HPLC on a C-8 column followed by derivatization with naphthalenedialdehyde and cyanide for 10 min at pH 9.0. The fluorescent derivatives of AdoMet and AdoHcy are then chromatographed by HPLC on a C-18 column monitored with a fluorescence monitor. The formation of the isoindole goes to 85% completion and the overall recovery of standards added to plasma is about 65%. Correction for recovery is made by addition of known quantities of AdoMet and AdoHcy to plasma. Normal values (+/- SE) for AdoMet were 102.7 nM +/- 9.9 and for AdoHcy were 22.7 +/- 3.1. PMID- 9866681 TI - Microquantification of cellular and in vitro F-actin by rhodamine phalloidin fluorescence enhancement. AB - Based on the enhancement of rhodamine phalloidin fluorescence after its binding to actin filaments we have developed a technique to quantify F-actin, drastically (>> 100 times) reducing consumption of the expensive fluorescent dye and sample material in comparison to previous methods. Depolymerization of F-actin is prevented by utilizing short incubation times and stabilization of the filaments by actin-binding proteins or formaldehyde. Equilibrium and kinetic mathematical models relating rhodamine fluorescence with F-actin concentrations were used to predict the optimal assay conditions. The method has been applied to measure relative and absolute F-actin concentrations in cytosolic fractions and stimulus induced actin polymerization in neutrophils. The cells were lysed with octy1-beta D-glucopyranoside, which is compatible with the assay due to its high critical micelle concentration. As the assay takes less than 1 h and eliminates all previously required washing or extraction steps, it is faster and much simpler than any other presented up to now for quantification of filamentous actin. Moreover, the method is unique for reliable and easy F-actin measurements in cell free systems. PMID- 9866682 TI - Rotating disk electrode voltammetric measurements of dopamine transporter activity: an analytical evaluation. AB - Rotating disk electrode (RDE) voltammetry at glassy carbon electrodes in 300- to 500-microL volumes has been shown in the literature by this laboratory and others to be useful in measuring dopamine and norepinephrine inward transport and release in preparations from rat brain and in expression systems. However, an analytical evaluation of the technique has not been made, and it is presented here in studies in striatal suspensions and human embryonic kidney cells expressing the human transporter for dopamine. The RDE was found to be able to measure dopamine and its metabolites with linear responses over the range of physiologically relevant concentrations with practical limits of quantification in the 10 to 50 nM range without signal and/or signal to noise enhancements. Response times of the technique were found to vary between 20 and 60 ms depending on rotation rate. Release and inward transport velocities were shown to be kinetically resolved. The glassy carbon electrode was found to be useful for several years at physiological temperatures without significant changes in electrochemical surface area, residual current, or response factors. Some sources of error in the measurement of release and inward transport values of velocity were noted and described. PMID- 9866683 TI - Kinetic analysis of enzyme inactivation under second-order conditions by use of substrate-to-product progress curves: application to the inhibition of trypsin by alpha-1 proteinase inhibitor. AB - The inhibition of bovine pancreatic trypsin by human alpha-1 proteinase inhibitor (alpha-1 PI) was studied under second-order conditions by continuously monitoring the fluorescence change due to the enzymatic hydrolysis of N alpha-benzoyl-L arginine 7-amido-4-methyl-coumarin as substrate. Employing equimolar starting concentrations of enzyme and inhibitor (110-220 nM), the fluorescence progress curve was analyzed according to the equation Pt = (kcat[S]/kiKm)In[ki[E]0t + 1], where ki is the second-order rate constant for the reaction, E + alpha-1 PI-->E* alpha-1 PI (inactive). ki was found to be 1.8 +/- 0.16 x 10(7) M-1 min-1 (at pH 7.0 and 25 degrees C), in close agreement with results obtained by alternative kinetic methods. The method reported appears to be valid and should be useful in the study of fast reactions where one of the reaction partners is an enzyme. An extension of the second-order progress curve approach to cover unequimolar mixtures of E and I is also offered. PMID- 9866684 TI - Contribution of component monosaccharides to the coordinates of neutral and sialyl pyridylaminated N-glycans on a two-dimensional sugar map. AB - High-performance liquid chromatography elution data on an amide-adsorption and a reverse-phase column, expressed in glucose units, of pyridylamino N-glycans have been analyzed with a new approach using multiple regression to obtain parameters for the contribution ascribable to each of 54 monosaccharide units. Our calculation was based on the 417 different N-glycan structures determined empirically. Depending on the increase in the amount of elution data, we got good correlation (r = 0.9998 for amide-silica and r = 0.9974 for octadecylsilica) and agreement between the observed and the calculated N-glycan elution coordinate values which correspond to the sum of the unit contribution of the component monosaccharides. These calculated values of unit contribution are useful in predicting glycan structure from an observed glucose unit on the map as well as to assume a glucose unit from a given structure. As an example of the application of the unit contribution values to the estimate of a sialyl N-glycan structure, the case of trisialyl triantennary N-glycans is described. PMID- 9866685 TI - Determination of the binding constant of a protein kinase C substrate, NG(28-43), to sodium dodecyl sulfate via the diffusion coefficient measured by pulsed field gradient nuclear magnetic resonance. AB - The binding affinity of a protein kinase C substrate, neurogranin peptide NG(28 43), to a sodium dodecyl sulfate micelle was analyzed quantitatively by the diffusion coefficient (Da) of the peptide determined by pulsed field gradient NMR. By use of a two-state model, the fraction of the peptide in the bound state, and hence the binding constant, can be estimated. The obtained binding constant is within the same order of magnitude as those reported for similar systems using other techniques. The present method may be generalized to measure the formation constants of other peptide:micelle complexes. PMID- 9866686 TI - Analysis of prostaglandin G/H synthase-2 inhibition using peroxidase-induced luminol luminescence. AB - The inducible form of the heme-protein prostaglandin G/H synthase (PGHS-2 or COX 2) has been established as a pivotal enzyme in the cascade of events leading to inflammation, hyperalgesia, and pyresis and represents a major therapeutic target in inflammatory disease. Accordingly, we have exploited the heme-catalyzed hydroperoxidase activity of recombinant hCOX-2 to generate luminescence in the presence of luminol, or a cyclic naphthalene hydrazide, and the substrate arachidonic acid. Arachidonate-induced luminescence was shown to be an index of real-time catalytic activity and demonstrated the turnover inactivation of the enzyme. Luminol luminescence was proportional to hCOX-2 concentration and gave accurate Km determinations for arachidonate. Inhibition of hCOX-2 activity, measured by luminescence, by a variety of selective (for COX-2) and nonselective inhibitors showed rank orders of potency similar to those observed with other in vitro and whole cell methods using the recombinant protein. The sensitivity of the luminescence assay also allowed determination of inhibitor potency at substrate concentrations below Km, distinguishing competitive inhibitors such as ibuprofen from time-dependent inhibitors such as DuP-697. Finally the use of higher quantum-yielding luminol analogues allowed measurement of cyclooxygenase activity at extremely low substrate and protein concentrations, enabling a variety of novel assay formats. PMID- 9866687 TI - Mass spectrometric sequencing of site-specific carcinogen-modified oligodeoxyribonucleotides containing bulky benzo[a]pyrene diol epoxide deoxyguanosyl adducts. AB - Site-specific carcinogen-modified oligonucleotides are often used in site directed mutagenesis and other biological and biochemical studies of structure function relationships. Postsynthetic analysis and confirmation of the sites of carcinogen binding in such oligonucleotides is an important step in the characterization of these site-specific carcinogen-DNA adducts. It is shown here that negative ion mode electrospray tandem mass spectrometry methods and collision-induced dissociation offer a rapid and convenient approach for the sequencing of products derived from the reaction of the carcinogenic and mutagenic metabolite of benzo[a]pyrene, the diol epoxide r7,t8-dihydroxy-t9,10 epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE), with the 11-mer oligonucleotide d(CATGCGGCCTAC). The site of reaction of anti-BPDE with either one of the three dG residues in this oligonucleotide can be accurately established by comparing the mass/charge ratios of the observed collision-induced dissociation fragments with calculated values. PMID- 9866688 TI - Fluorometric detection of paralytic shellfish poisoning toxins. AB - A rapid qualitative screening method was developed for the fractionation of paralytic shellfish poisoning toxins. Periodic acid, t-butyl hydroperoxide, and hydrogen peroxide were tested as oxidants for the fluorometric detection of paralytic shellfish toxins. Hydrogen peroxide was found to be the most convenient and efficient oxidant since the fluorescence can be detected after the incubation of toxins at 100 degrees C for 3-5 min. In addition to the structure of the compound, the incubation temperature and time, the amount of acid, and the peroxide concentration affect the fluorescence reaction. This method was more efficient than the previously published peroxidation methods which involved lengthy incubation periods or time-consuming pH adjustment. Also, far greater sensitivity was achieved with the new method with levels of 0.027, 0.054, 0.023, 0.003, 0.0002, and 0.0006 pmol being easily detected for saxitoxin, neosaxitoxin, gonyautoxin 1 and 4, gonyautoxin 2 and 3, C toxins, and B toxins, respectively. The method is particularly valuable for the screening of fractions separated by column chromatography. PMID- 9866689 TI - Selective bridging of bis-cysteinyl residues by arsonous acid derivatives as an approach to the characterization of protein tertiary structures and folding pathways by mass spectrometry. AB - Bis-cysteine selective modifications were successfully applied with melarsen oxide (MEL), an arsonous acid derivative, for tertiary structural studies of peptides and a model protein. The arsonous acid modified peptides and proteins were amenable to direct characterizations by mass spectrometry, e.g., direct molecular weight determinations and mass spectrometric peptide mapping that identified stoichiometry and sites of modification, respectively. Proteolytic digestion and mass spectrometric fragmentation of modified oxytocin showed that MEL-bridged peptide derivatives are structural homologues to the disulfide-bonded macrocyclic peptides. Mass spectrometric analyses determined the MEL modification site in partially reduced and selectively modified bovine pancreatic trypsin inhibitor (BPTI) bridging Cys-14 and Cys-38. The BPTI.MEL derivative was resistant to proteolysis by both Lys-C and trypsin and thus represented a rigid structure like native BPTI. MEL exhibited several advantageous features such as (i) cross-linking two closely spaced thiol groups, providing detailed tertiary structure information; (ii) high solubility as monomeric ortho acid in aqueous and organic solutions; (iii) adding a relatively large mass increment to proteins upon single modification; (iv) enabling UV monitoring of the derivatization due to a strong chromophor; and (v) performing fast and specific modifications of bis thiol groups in proteins to form stable structures without any side reactions even with a high molar excess of MEL. The investigated physical and chemical properties of MEL suggest general applicability for selective bis-thiol modifications, enabling protein structure-function studies in both soluble and membrane proteins and the study of protein-folding reactions. PMID- 9866690 TI - Identification of multidrug resistant protein 1 of mouse leukemia P388 cells on a PVDF membrane using 6-aminoquinolyl-carbamyl (AQC)-amino acid analysis and World Wide Web (WWW)-accessible tools. AB - Multidrug resistant protein 1 (MDR1) in a doxorubicin-resistant mouse leukemia cell line (P388/DOX) was identified using its amino acid composition combined with protein database searching (ExPASy and EMBL PROPSEARCH) via the World Wide Web. The proteins were separated by one-dimensional SDS-polyacrylamide gel electrophoresis, blotted onto a polyvinylidene fluoride membrane, and stained with Coomassie brilliant blue. A 160-kDa protein band was acid-hydrolyzed in the vapor phase (6 N HC1) and converted to 6-aminoquinolyl-carbamyl (AQC)-amino acids without extraction of the amino acids from the membrane. The amino acid composition of the protein was determined using the sensitive AQC-amino acid analysis method, improving our previously described method. The improved method involved using a Cosmosil 5C8-MS column instead of a Pegasil C8; replacement of the mobile phase A, constituent, 75 mM ammonium phosphate (pH 7.5), with 30 mM sodium phosphate buffer (pH 7.2); and slight modification of the separation program (9). All manipulations for protein hydrolysis and AQC derivatization were carried out in a hood using clean tools. This minimized contamination of amino acids at the low femtomolar level. A database search was carried out with bovine serum albumin as a calibration protein. MDR1 in P388/DOX was ranked first by both databases with high reliability (score 14 for ExPASy, distance 1.34 for EMBL). PMID- 9866691 TI - A continuous spectrophotometric assay for thymidine and deoxycytidine kinases. AB - We have developed a continuous spectrophotometric assay for thymidine and deoxycytidine kinase activities by coupling nucleoside 5'-monophosphate formation to a methylation reaction which generates a product absorbing at 340 nm. With thymidine kinase, we used the alternate substrate deoxyuridine and coupled the reaction to thymidylate synthase. For deoxycytidine kinase, we coupled the reaction to a thymidylate synthase mutant which converts the product 2' deoxycytidine-5'-monophosphate (dCMP) to m5dCMP. In both cases, the methylation reactions are accompanied by conversion of 5,10-methylene-5,6,7,8 tedrahydrofolate to 7,8-dihydrofolate and can be continuously monitored by the increase of absorbance at 340 nm. The assay should be particularly useful for kinetic studies, and for the purification of these enzymes from various sources. PMID- 9866693 TI - Negative cooperativity in tryptophan synthase alpha subunit dissociation is caused by the bound coenzyme: pyridoxal 5'-phosphate. AB - Sedimentation equilibrium studies of dilute solutions of tryptophan synthase reveal dissociation from the holoenzyme form, alpha 2 beta 2, into mixtures of alpha beta 2, small amounts of beta 2, and alpha as well as the original alpha 2 beta 2 holoenzyme. The holoenzyme form is stabilized by pyridoxal 5'-phosphate. A new sedimentation equilibrium analytical procedure shows the dissociation of the second alpha subunit to be negatively cooperative. The analytical procedure calculates theoretical error profiles with assumed values of the dissociation constant, k, and a cooperativity parameter until a match is made between one of the theoretical profiles and that computed from experimental data. The latter profile is calculated with an experimentally determined k and assumed values of the cooperativity parameter. PMID- 9866692 TI - Measurement of photosystem I activity with photoreduction of recombinant flavodoxin. AB - Flavodoxin can function as an alternative electron acceptor for photosystem I (PSI) in place of ferredoxin under iron-limiting conditions. The isiB gene, encoding the flavodoxin in Synechococcus sp. PCC 7002, was overexpressed in Escherichia coli. Under the conditions employed, most recombinant flavodoxin (rFlvd) was in soluble form with cofactor correctly inserted. The absorption spectrum of rFlvd was identical to that of the native flavodoxin of the cyanobacteria. Photoreduction of rFlvd by PSI particles and thylakoid membranes was determined directly by monitoring the absorption change at 467 nm. The optimal conditions for rFlvd photoreduction were determined. Compared to other methods currently employed to measure PSI activity such as oxygen uptake in the presence of methyl viologen and NADP+ photoreduction in the presence of ferredoxin and ferredoxin:NADP+ oxidoreductase, measurement of PSI activity with flavodoxin as an electron acceptor has several advantages. It measures the full chain electron transfer chain of PSI since flavodoxin accepts electrons from FA/FB and it is much simpler than the method with NADP+ photoreduction. With this method, we found that the affinity of wild-type PSI for rFlvd was 35% higher than that of the PsaE-less PSI, showing that this method is sensitive to structural changes of PSI. Our results demonstrate that rFlvd photoreduction is an effective and simple method for PSI activity measurement. PMID- 9866694 TI - Nonradioactive Northern blot analysis of plant RNA and the application of different haptens for reprobing. AB - Northern blot analysis using radioactive probes is still the most common technique to determine the accumulation of transcripts in cells and tissues. The main disadvantages of this technique are the possible health hazard, inconvenience during handling, the high amount of RNA target necessary for detection, and difficulties with stripping and reprobing. In this paper, we propose an easily applicable protocol for Northern blot analysis of plant RNA with enhanced sensitivity using digoxigenin-, fluorescein-, or biotin-labeled in vitro transcripts derived from PCR products. Furthermore, we show the rehybridization of Northern blots with fluorescein as a second hapten, avoiding any stripping procedure. PMID- 9866695 TI - Ascorbate increases electrotransformation efficiency of intact maize cells. PMID- 9866696 TI - An immunoaffinity procedure for determination of enzyme activity at elevated temperatures. PMID- 9866697 TI - Generation of multiple site-specific mutations in a single polymerase chain reaction product. PMID- 9866698 TI - A combined flow injection-chemiluminescent method for the measurement of radical scavenging activity. PMID- 9866699 TI - Vancomycin resistance in Enterococcus faecium: a capillary electrophoresis-based assay for VanX enzyme. PMID- 9866700 TI - Mode of adsorption and orientation of an extracellular matrix protein affect its cell-adhesion-promoting activity. AB - Cell adhesion to extracellular matrix contributes to the organization of tissues and modulates cell behavior. In conventional cell adhesion assays, plastic wells are coated with matrix proteins and assayed for their adhesion-promoting activity. We show here that factors such as sample composition, coating buffers, and manufacturers' plastic treatment markedly affect cell adhesion to the extracellular matrix protein laminin-5 (Ln-5). These factors were shown to affect adsorption efficiency as determined by measuring total adsorbed protein with a polyclonal anti-Ln-5 antiserum. They also influence the availability of the epitope for an adhesion-blocking anti-Ln-5 monoclonal antibody, suggesting that coating conditions affect the orientation of Ln-5. Generally, cell adhesion correlates more strongly with the availability of the epitope for the adhesion blocking antibody than with total adsorbed Ln-5. Our data further indicate that cell adhesion to other matrix proteins may be influenced by similar factors. Adding Ln-5 samples to plastic wells that had been precoated with non-adhesion blocking anti-Ln-5 antibodies made cell adhesion independent of factors such as sample composition, coating buffers, and source of plastic. Thus, the control of adsorption efficiency and orientation of extracellular matrix proteins is essential for creation of reliable and reproducible conditions in cell adhesion assays. PMID- 9866701 TI - Evaluation of methods for the quantitation of cysteines in proteins. AB - Several methods for the quantitation of cysteines in proteins have been evaluated and compared. Titration of protein sulfhydryl groups with 5,5'-dithiobis(2 nitrobenzoate) (DTNB) under carefully controlled conditions has extended the detection limits of this method with high accuracy and reproducibility. Results are reported for a variety of enzymes containing a range of total cysteines with different degrees of solvent accessibility and reactivity. A papain amplification assay has also been examined, in which reactivation of the disulfide-blocked active site cysteine of papain can be achieved by a coupled reaction with protein sulfhydryl groups. Detection of sulfhydryls by this amplification assay can be extended, by increasing the enzyme assay times, to achieve over a 40-fold increase in sensitivity over the improved DTNB titration method. Alternatively, titration of enzyme cysteinyl residues with either bromobimane or a maleimide derivative of naphthopyranones has the advantage that a fluorescent product results upon modification of the sulfhydryl group. Reaction of bromobimane with several different enzymes results in nonspecific background fluorescence that limits the detection range of this method unless the products are separated. In contrast, low background fluorescence and high quantum yields with maleimide naphthopyranoses has allowed detection of protein cysteinyl residues with very high sensitivities. PMID- 9866702 TI - 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride as a fluorescent labeling reagent for determination of amino acids in high-performance liquid chromatography and its application for determination of urinary free hydroxyproline. AB - A highly sensitive fluorescent labeling reagent, 4-(5,6-dimethoxy-2 phthalimidinyl)-2-methoxyphenylsulfonyl chloride (DMS-Cl), for determination of amino acids by HPLC has been developed. DMS-Cl reacted with amino acids in the basic medium to produce the corresponding fluorescent sulfonamides (excitation and emission wavelength: 318 and 406 nm in aqueous acetonitrile, respectively). When the reactivity of DMS-Cl was investigated by means of reversed-phase HPLC using hydroxyproline (Hyp) and alanine (Ala) as model compounds, the reaction of Hyp was completed within 5 min at 25 degrees C and that of Ala within 15 min at 70 degrees C. The efficiency of conversion of Hyp into the fluorescent derivative was about 100%. The detection limits (signal-to-noise ratio = 3) of almost all amino acids were less than 5 fmol/injection. When the concentration of urinary free Hyp was measured with HPLC using DMS-Cl, the concentration (mean +/- SD, n = 10) was 3.8 +/- 2.76 microM (2.7 +/- 1.71 nmol/mg creatinine). PMID- 9866703 TI - High-throughput screening assay for helicase enzymes. AB - We have developed a novel 96-well microtiter plate high-throughput screening filtration assay for the detection of helicase activity. In this paper we present data for detection of helicase activity of the UL5/8/52 helicase-primase complex from herpes simplex virus 1 (HSV1). The assay involves the detection of radioactively labeled oligonucleotide annealed to a single-stranded circular DNA following capture of the annealed complex on silica beads. We have screened over 200,000 samples containing small organic molecules and natural products and identified T157602, a two-amino thiazole, as a specific inhibitor of HSV replication. PMID- 9866704 TI - An enzyme-linked immunosorbent assay for the detection of agents which interfere with the DNA binding activities of transcription factors--exemplified by NF-IL6. AB - Binding of transcription factors to DNA is usually detected by the electrophoretic mobility shift assay also known as gel-shift or band-shift assay. Recently the use of biosensors has allowed factor/DNA interactions to be followed in real time. However, neither of these approaches lends itself easily to high throughput screening of agents which might interfere with this process. We have therefore developed a 96-well plate-based enzyme-linked immunosorbent assay for this purpose. Biotinylated oligonucleotides bound to streptavidin-coated plates are used to capture recombinant transcription factor proteins which can then be detected using specific antibodies which in turn are recognised by peroxidase conjugated antisera. The peroxidase catalyzes conversion of a colorless substrate to a colored product which can be quantified by optical densitometry on a plate reader. Agents which interfere with the binding of the transcription factor to DNA reduce the optical density in the well. Using the NF-IL6 transcription factor we show that the assay can detect reductions to 86% at significant levels. The assay may be readily adapted for robotic manipulation, making it ideal for high throughput screening. PMID- 9866706 TI - A gas chromatographic method for the sugar analysis of 3,6-anhydrogalactose containing algal galactans. AB - This paper describes a new gas chromatographic method for the compositional analysis of algal galactans containing 3,6-anhydrogalactose, which is readily destroyed in usual acid hydrolysis and methanolysis. Mercaptolysis of galactans was carried out under an anhydrous condition using the newly developed solvent system, an ethanethiolic HCl solution which had been prepared by blowing dry HCl gas into ethanethiol. By heating galactans in 0.5 N HCl/ethanethiol at 60 degrees C for 12 h, component sugars including 3,6-anhydrogalactose were released quantitatively as their diethyl mercaptals. The monosaccharide diethyl mercaptals were trimethylsilylated and analyzed by gas-liquid chromatography. Each component monosaccharide was detected as a single peak since neither anomeric nor structural isomers can be produced by mercaptolysis. Furthermore, the solvent system was improved by the addition of methanol in a ratio (2/1, v/v) of ethanethiol to methanol. Heating of galactans at 60 degrees C in 0.5 N HCl/[ethanethiol:methanol (2/1, v/v)] released more rapidly and quantitatively monosaccharide diethyl mercaptals without any formation of monosaccharide methyl glycosides. The present method was applicable to not only algal galactans but also algal powders. PMID- 9866705 TI - Carbonothioate phospholipids as substrate for a spectrophotometric assay of phospholipase A2. AB - A continuous spectrophotometric assay for phospholipase A2 (PLA2) was developed using novel carbonothioate phospholipids. These phospholipid analogues contain a carbonothioate bond in the place of the sn-2 ester of the natural substrates of phospholipase A2 and were synthesized in a one-pot two-step reaction. Phospholipase A2 from cobra venom (Naja naja atra) hydrolyzes carbonothioate phospholipids and liberates a free thiol, alkylmercaptan, which is reacted with 5,5'-dithiobis(2-nitrobenzoic acid) to yield a product that absorbs at 412 nm. The kinetic studies on PLA2 hydrolysis of carbonothioate phospholipids were carried out in pure phospholipid forms and in Triton X-100 mixed micelles. The hydrolysis of pure carbonothioate phospholipids exhibits an interfacial activation phenomenon. The hydrolysis of phospholipid in mixed Triton X-100 micelles follows classical Michaelis-Menten kinetics. In a mixed micellar system, the catalytic efficiency observed with this series of substrates is two orders of magnitude lower than that of the hydrolysis of the natural substrate dipalmitoyl phosphocholine. However, these substrates bind to the enzyme over 10 times tighter than does the natural substrate. Application of this carbonothioate assay to screen both reversible and irreversible enzyme inhibitors of phospholipase A2 is also demonstrated. PMID- 9866707 TI - Enzymatic method for the simultaneous determination of hyaluronan and chondroitin sulfates using high-performance liquid chromatography. AB - A chromatographic method for the simultaneous determination of hyaluronan and chondroitin sulfates was examined. Hyaluronan differs from chondroitin sulfates in the susceptibility to chondroitinase ABC under alkaline conditions. When hyaluronan and chondroitin sulfates were treated with chondroitinase ABC in the buffered solution (pH 9.1), chondroitin sulfates were selectively degraded to the unsaturated disaccharides, whereas hyaluronan was not. Subsequently, hyaluronan in the reaction mixture was digested to the unsaturated tetrasaccharide and hexasaccharide at pH 6.0 by Streptomyces hyaluronidase in the presence of zinc ion (inhibitor for chondroitinase ABC). The separation of the resulting unsaturated disaccharides from chondroitin sulfates and the unsaturated oligosaccharides from hyaluronan was achieved by a reversed-phase ion-pair HPLC. The structural polydispersities of hyaluronan and chondroitin sulfates in the molecular weight, the sulfation position, or the components of uronic acid did not affect their determination. The usefulness of the present method was proved by application to the porcine skin samples. PMID- 9866708 TI - Analysis of cytochrome P450 metabolites of arachidonic and linoleic acids by liquid chromatography-mass spectrometry with ion trap MS. AB - We have used reversed phase-high performance liquid chromatography-mass spectrometry (RP-HPLC-MS) with an ion trap mass spectrometer to study the metabolism of arachidonic and linoleic acids by human recombinant cytochrome P450 (CYP) enzymes. We first recorded the MS2 spectra of the carboxylate anions of epoxides, diols, omega-side chain, and bisallylic hydroxy fatty acids of arachidonic, octadeuterated arachidonic, and linoleic acids. The metabolites formed by CYP2C9 and CYP2C19 were then studied. CYP2C9 converted arachidonic and linoleic acids to epoxides/diols and monohydroxy fatty acids. Some hydroxyeicosatetraenoic acids (HETEs) were studied in detail to investigate the oxygenation mechanism. Incubation of CYP2C9 under oxygen-18 gas showed that all HETEs had incorporated oxygen-18 to the same degree. Chiral HPLC showed that CYP2C9 formed 15R-HETE (72% of the R enantiomer), 13S-HETE (90%), and 11R-HETE (57%). RP-HPLC-MS analysis revealed that CYP2C19 oxygenated arachidonic acid to 19-HETE, 14,15-epoxyeicosatrienoic acid (EET), and 8,9-EET as main metabolites. The method was sufficiently sensitive to identify arachidonic acid metabolites formed by some other isozymes. RP-HPLC-MS with MS2 seems to be useful for rapid identification of fatty acid metabolites in complex mixtures formed by cytochrome P450. PMID- 9866709 TI - Biotinylated enzyme inhibitorsorbent assay: a specific method for quantitating enzyme and its inhibitor. AB - A biotinylated enzyme inhibitorsorbent assay (BEISA) for quantitating enzyme and its inhibitor has been developed. The assay is based on the competition between unlabeled enzyme and biotin-labeled enzyme for binding by an immobilized inhibitor or between free inhibitor and the immobilized inhibitor for binding by a biotin-labeled enzyme followed by reaction of the biotin-bound complex with a streptavidin-alkaline phosphatase conjugate. The amount of enzyme or inhibitor can be determined from the intensity of color produced by the alkaline phosphatase acting on its substrate. Trypsin and its inhibitor from egg white (ovomucoid) were used as a model for the BEISA. The results indicated that the BEISA is a simple, sensitive, and specific method that can be used to quantitate the amount of an enzyme or its inhibitor and it is amenable to high-throughput analysis and automation. The BEISA can be also applied to any enzyme that has an appropriate inhibitor. PMID- 9866710 TI - Iron quantification in cerebrospinal fluid. AB - Several studies have attempted to measure iron levels in cerebrospinal fluid (CSF), which is believed to contain very low levels of iron. In general, the assays used in these studies suffered from poor reproducibility or lack of sensitivity. In the present study, an assay was developed that enables iron quantitation in CSF using 150 microliters of undiluted CSF, sample digestion, dot blotting, Perls' histochemistry, 3,3'-diaminobenzidine enhancement, and densitometry. Inter- and intraassay variability was low and sensitivity high (279 pg iron in 100 microliters). There were negligible readings for standard curves utilizing copper in place of iron. The iron content in eight normal CSF samples was 61.01 +/- 18.3 (SD) micrograms/liter, and in nine normal sera it was 1332.7 +/- 408.2 (SD) micrograms/liter. The present assay enables reliable measures of iron levels in biological samples. This assay should enable future studies that address how CSF iron levels change during the course of various disease states. PMID- 9866711 TI - Determination of binding constants by equilibrium titration with circulating sample in a surface plasmon resonance biosensor. AB - A commercial surface plasmon resonance biosensor, BIACORE X, is employed as a detector in a closed loop of a small sample volume. The sample is continuously circulated by an external syringe pump over two sensor spots, one functionalized with immobilized binding sites to a soluble binding partner in the mobile phase and one serving as a reference surface. A binding isotherm for the interacting macromolecules can be obtained by a stepwise titration of the soluble reactant into the circulating loop, each step followed by observation of the signal increase until equilibrium is attained. Binding constants can be measured under conditions free of mass transport artifacts and without the requirement for regeneration of the immobilized binding sites. This procedure is similar to the stepwise titration procedure described for the cuvette-based sensor design (D. R. Hall and D. J. Winzor, 1997, Anal. Biochem. 244, 152-160). In the presented configuration, the high baseline stability of the instrument combined with the availability of a reference surface for the detection of nonspecific binding permits refractive index changes upon addition of the aliquots to be measured, as well as accounting for temperature or instrumental drifts, and allows for a very long experimental time. This feature extends the applicability of equilibrium titration to systems with higher affinity or slower dissociation rate constants. Furthermore a solution competition titration is described that avoids artifacts from the immobilization procedure to provide a method for measurement of binding constants in solution. Kinetic information on the complex dissociation can also be obtained by combination of sample delivery via the external pump with the injection of competitor via the microfluidics of the biosensor. The rapid injection of high concentrations of competitor allows the observation of fast dissociation processes under conditions minimizing rebinding. PMID- 9866712 TI - Colorimetric determination of citrulline residues in proteins. AB - A method is described for the direct colorimetric determination of citrulline residues in proteins based on the reaction with diacetylmonoxime in the presence of lower concentrations of sulfuric acid. The reduced sensitivity due to the lower acid concentration was overcome by the addition of ferric chloride which also contributed to the color stabilization. Insoluble proteins or proteins resulting in turbidity can be analyzed following partial hydrolysis of those with enzyme or acid. The molar absorption coefficient (epsilon) for citrulline at 464 nm was 2.8 x 10(4). The method enabled us to determine low levels of protein bound citrulline which are beyond the limitations of conventional methods using an amino acid analyzer. We determined the citrulline contents in the cornified cells of the epidermis of newborn rats, as well as soybean trypsin inhibitor. PMID- 9866714 TI - Strategic biochemical analysis of mucins. AB - MUC-type mucins comprise a family of structurally related molecules, which are expressed in epithelia of the body that are in close contact with the milieu. Because of their large sizes and very complex structures, containing very extensive O-glycosylation, MUC-type mucins are difficult to study by conventional techniques. Many see MUC-type mucins as protective molecules; however, functional studies on the individual MUC-type mucins are very scarce. At present, essential steps in MUC research are to characterize the specific expression patterns of each MUC-type mucin in the body and to find methods to reliably quantify these MUC-type mucins. These aims can only be met at the level of the primary sequences of the MUC-type mucins, as the O-glycosylation even within one species of MUC type mucin is not only very complex, but may also vary among individuals, organs, and cell types. We will discuss some recent advances in mucin research, particularly the identification of MUC precursor molecules in metabolic labeling experiments. We will try to define some strategic considerations in the study of the expression patterns of MUC-type mucins, which circumvent the complications caused by the very complex and heterogeneous O-glycosylation of the molecules. PMID- 9866713 TI - Determination of proteins, phosphatidylethanolamine, and phosphatidylserine in organic solvent extracts of tissue material by analysis of phenylthiocarbamyl derivatives. AB - Amino acid analysis of organic solvent extracts of tissue material has been evaluated for determination of protein content. Conventional ninhydrin-based analysis does not allow determination of a large number of lipid-rich samples. Therefore, the hydrolyzed samples were treated with phenylisothiocyanate and the phenylthiocarbamyl (PTC) derivatives obtained were separated by reverse-phase HPLC. With this method, analysis of many lipid-rich samples is feasible. In addition, phosphatidylethanolamine and phosphatidylserine can then be determined together with the amino acid constituents. The PTC/reverse-phase HPLC method was used for analysis of chloroform/methanol extracts of spinal cord, lung, and bile after chromatography on Lipidex 5000 in methanol/ethylene chloride, 4:1 (v/v). The chromatography profiles show that in all tissue samples the proteins elute before the phospholipids. Consequently, a single step of Lipidex 5000 chromatography can be used to purify polypeptides present in organic solvent extracts. Using pulmonary surfactant extracts (with about 98% phospholipids and 1 2% proteins), we find that individual contents of surfactant proteins B and C can be determined by amino acid analysis. PMID- 9866715 TI - Sodium-dependent amino acid transport is preserved in lyophilized reconstituted apical membranes from intestinal epithelium. AB - We demonstrate for the first time that functional electrogenic Na(+)-dependent amino acid transport is preserved for extended periods when purified brush border membranes prepared in hypotonic media are lyophilized and then rehydrated in buffer containing mannitol, NaSCN, and/or KSCN/valinomycin. Reconstituted lyophilized apical membranes from small intestine formed morphologically, physiologically, and thermodynamically normal vesicles which transported L alanine via system B into an osmotically active space energized by secondary active transport, as measured under equilibrium and nonequilibrium conditions. The lyophilized membranes are readily prepared and stored, thereby providing a means to pool large quantities of formed vesicles that are useful in examining cloned and reconstituted native amino acid transporter polypeptides. PMID- 9866716 TI - The yeast mitochondrial intermembrane space: purification and analysis of two distinct fractions. AB - We have developed a protocol for the sequential release of the intermembrane space (IMS) content of Saccharomyces cerevisiae mitochondria. Two distinct fractions were obtained: a soluble IMS with cytochrome b2 as key marker and a salt-extractable IMS with cytochrome c as key marker. The identity of several proteins was determined by amino-terminal amino acid sequencing. The IMS fractions were devoid of contaminations from cytosol and mitochondrial outer and inner membranes. By subtraction analysis, the protein profiles of soluble and salt-extractable IMS fractions were depleted of contaminating bands derived from matrix proteins. The fractionation method will provide the basis for the further analysis of IMS proteins and characterization of their functions in bioenergetics, mitochondrial biogenesis, and regulatory processes. PMID- 9866717 TI - A simplified gradient solvent delivery system for capillary liquid chromatography electrospray ionization mass spectrometry. AB - We describe a simple solvent delivery system for gradient capillary HPLC at nanoliter per minute flow rates. The novel aspect of the system is that solvents are delivered one at a time, using a switching valve, into a relatively large volume mixing chamber. Efficient mixing in the chamber causes the formation of a sigmoidal gradient from the initial solvent to the subsequent solvent, which is then delivered to a capillary column. The shape of the gradients formed can be predicted from a simple theoretical model. Gradients of different slope can be formed by varying either the size of the chamber or the system flow rate. The system is robust, reproducible, and simple to operate. We provide a detailed protocol of how to construct a low-cost capillary HPLC system consisting of two syringe pumps, a capillary mixing chamber, a capillary column, and a zero dead volume microelectrospray interface. We demonstrate that the coupling of this HPLC system to a mass spectrometer enabled us to identify proteins at the low femtomole level in solution-phase digests and at the picomole level in digests of samples separated on SDS-PAGE gels. We believe that the strategy presented will be useful as a general method for the characterization of proteins and peptides by capillary HPLC-electrospray mass spectrometry. PMID- 9866718 TI - Analysis of DNA adduct, S-[2-(N7-guanyl)ethyl]glutathione, by liquid chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry. AB - Sensitive and specific isotope dilution liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) methods were developed for the detection and quantitative analysis of S-[2-(N7 guanyl)ethyl]glutathione as a DNA adduct formed upon exposure of animals to carcinogenic 1,2-dihaloethanes. Separation and analysis were performed using microbore HPLC coupled in-line to an electrospray ionization triple quadrupole mass spectrometer. S-[2-(N7-guanyl)[2H4]-ethyl] glutathione was synthesized and used as internal standard. These methods provide structural confirmation of the adduct as well as quantitative analysis with the accuracy and precision necessary to measure biologically relevant levels in small tissue sample sizes (< 1 g). The sample detection limits in in vivo tissue extracts were 100 pg and 5 pg on-column for LC/MS and LC/MS/MS methods, respectively. Selected-ion monitoring mode was used to monitor the product ions of the doubly charged molecular ion. The application of these methods was demonstrated by measuring the DNA adduct levels in rat and fish samples after exposure to 1,2-dihaloethanes. The method has application in studies of DNA adduct formation as a biological marker of exposure to carcinogens and for environmental monitoring of 1,2-dihaloethanes. PMID- 9866719 TI - Oligosaccharide dehydrogenase-catalyzed assay for the determination of polysaccharides. AB - Oligosaccharide dehydrogenase (ODH), an enzyme known to have a broad selectivity for reducing sugars of low molecular weight, was investigated to determine its catalytic properties with larger polysaccharides. Six substrates were studied: pullulan standards with molecular weights of between 5,400 and 90,900, debranched starch, and dextran. In addition, maltotriose, isomaltotriose, maltose, and glucose were used as substrates for comparison. ODH catalyzed the oxidation of the large pullulans with a degree of polymerization of at least 560. Isomaltotriose and dextran were not oxidized. ODH activity for the pullulans, expressed as the rate constant Kps, was only three times lower than that for maltose. When the oxidation of sugars with ODH was coupled to a color-forming reaction, quantitative spectrophotometric determination of sugars was possible using either Meldola's blue or N-methylphenazinium as electron acceptors in combination with nitrotetrazolium blue. Linear calibration curves for maltose, maltotriose, and debranched starch were obtained using this ODH method and compared with curves from the conventional spectrophotometric copper sulfate method. This work demonstrates that ODH can be advantageously used for the determination of polysaccharides. PMID- 9866720 TI - Determination of folate patterns in mouse plasma, erythrocytes, and embryos by HPLC coupled with a microbiological assay. AB - Folates are important cofactors in one-carbon metabolism. Disturbances in folate homeostasis and metabolism may be related to an increased risk of cardiovascular disease and carcinogenesis and may lead to congenital malformations, namely neural tube defects. Determination of these compounds in biological samples is often a problem due to the existence of numerous folate metabolites, their relative instability, and the low contents in serum and most tissues. As existing methods have distinct limitations, we developed a method, which facilitates the separation as well as the sensitive detection of eight folates by coupling HPLC with a microbiological assay. After a simple sample preparation, including deproteinization and enzymatic hydrolysis of folate polyglutamates, extracts were chromatographed, fractions were collected on microtiter plates, and folates were quantitated using the Lactobacillus casei assay. The raw data were processed using a computing system after reconstructing the HPLC chromatogram with the bacterial growth data. Using the described method, the eight physiologically occurring folate monoglutamates could be simultaneously determined. The detection limits were 2-20 fmol per injection. The application of the method was demonstrated with the analysis of the folate pattern in milligram or sub milligram quantities of plasma, erythrocyte, and embryos of pregnant mice during organogenesis. PMID- 9866721 TI - Detection of metallothionein isoforms from three different species using on-line capillary electrophoresis-mass spectrometry. AB - An on-line capillary electrophoresis-mass spectrometry method (CE-MS) for the detection of metallothionein (MT) isoforms is described. The detected masses were usually within 1-1.5 mass units of the expected molecular weights. MT-containing samples from rabbit, sheep, and yeast (Saccharomyces cerevisiae) were subjected to CE-MS analysis. The analysis of rabbit liver MT revealed the masses of 10 proteins/peptides. Five of the detected masses corresponded well with the expected masses calculated from the amino acid sequence of previously described MT isoforms, one was suspected to be a deacetylated form of MT-2A, one was presumed to be a yet unknown isoform, and three masses were classified as non-MT compounds. From the analysis of a fetal sheep liver extract six proteins were detected of which three masses corresponded to previously described MT isoforms. Two purified MT subforms from S. cerivisiae (encoded by the CUP1 locus) were analyzed for their copper content and both forms were found to contain eight copper atoms per molecule. PMID- 9866722 TI - Gel electrophoretic quantitation of protein carbonyls derivatized with tritiated sodium borohydride. AB - A method for the quantitation of protein carbonyls, which have been widely employed as markers of protein oxidative damage, is described. Protein carbonyls were derivatized with tritiated sodium borohydride and the tritiated proteins were separated on SDS-PAGE. Protein bands, visualized by Coomassie blue staining, were then excised and incubated in 30% H2O2 at 60 degrees C for 48 h. Tritium, incorporated into the proteins, was quantitated by liquid scintillation counting after gel solubilization by H2O2. This method can be applied to the measurement of carbonylation of specific proteins as it employs SDS-PAGE and has the advantage that unreacted NaB3H4 in the labeling reaction mixture need not be removed. The present method, when combined with immunochemical detection of protein carbonyls, should be very useful in the quantitation of oxidative damage to individual proteins. PMID- 9866723 TI - Phenol-extracted plant proteins can be renatured and assayed in gel for protein kinase activity. PMID- 9866724 TI - A method for obtaining reporter gene activity and nuclear extracts simultaneously from transiently transfected cells. PMID- 9866725 TI - Problems with phosphoamino acid analysis using alkaline hydrolysis. AB - Although a seemingly simple concept, sample volume and the reaction vessel size are important considerations when undertaking an alkaline hydrolysis of a phosphoprotein, particularly if the phosphoamino acid of interest is either phosphohistidine or phosphotyrosine. It should be noted that the experiments conducted in this article used large concentrations of both phosphotyrosine and the most stable form of phosphohistidine (8), which highlights the problems that may be faced during phosphoamino acid analysis of a protein sample that contains only small amounts of either phosphoamino acid. Although not tested, it is likely that similar hydrolysis effects may occur for phospholysine. If the reaction volume is to be kept to a minimum and the alkaline digestion is from either a membrane blot or in solution, then the use of a mineral oil overlay should be considered to prevent concentration of the alkali and hydrolysis of the phosphate moiety. PMID- 9866726 TI - Analysis of subunit structures of proteins by polyacrylamide gel electrophoresis. PMID- 9866727 TI - Emerging infectious diseases--South Africa. PMID- 9866728 TI - Quasispecies structure and persistence of RNA viruses. AB - Viral quasispecies are closely related (but nonidentical) mutant and recombinant viral genomes subjected to continuous genetic variation, competition, and selection. Quasispecies structure and dynamics of replicating RNA enable virus populations to persist in their hosts and cause disease. We review mechanisms of viral persistence in cells, organisms, and populations of organisms and suggest that the critical interplay between host and viral influences (including in some cases the quasispecies organization) is the main driving force for long-term survival of viruses in nature. PMID- 9866729 TI - Ecologic studies of rodent reservoirs: their relevance for human health. AB - Within the past few years, the number of "new" human diseases associated with small-mammal reservoirs has increased dramatically, stimulating renewed interest in reservoir ecology research. A consistent, integrative approach to such research allows direct comparisons between studies, contributes to the efficient use of resources and data, and increases investigator safety. We outline steps directed toward understanding vertebrate host ecology as it relates to human disease and illustrate the relevance of each step by using examples from studies of hosts associated with rodent-borne hemorrhagic fever viruses. PMID- 9866730 TI - Diphtheria in the former Soviet Union: reemergence of a pandemic disease. AB - The massive reemergence of diphtheria in the Newly Independent States of the former Soviet Union marked the first large-scale diphtheria epidemic in industrialized countries in 3 decades. Factors contributing to the epidemic included a large population of susceptible adults; decreased childhood immunization, which compromised what had been a well-established childhood vaccination program; suboptimal socioeconomic conditions; and high population movement. The role of a change in the predominant circulating strains of Corynebacterium diphtheriae in this epidemic remains uncertain. Massive, well coordinated international assistance and unprecedented efforts to vaccinate adults were needed to control the epidemic. PMID- 9866732 TI - Rotavirus. AB - Rotavirus, the most common diarrheal pathogen in children worldwide, causes approximately one third of diarrhea-associated hospitalizations and 800,000 deaths per year. Because natural infection reduces the incidence and severity of subsequent episodes, rotavirus diarrhea might be controlled through vaccination. Serotypespecific immunity may play a role in protection from disease. Tetravalent rhesus-human reassortant rotavirus vaccine (RRV-TV) (which contains a rhesus rotavirus with serotype G3 specificity and reassortant rhesus-human rotaviruses with G1, G2, and G4 specificity) provides coverage against the four common serotypes of human rotavirus. In clinical trials in industrialized countries, RRV TV conferred 49% to 68% protection against any rotavirus diarrhea and 61% to 100% protection against severe disease. This vaccine was licensed by the U.S. Food and Drug Administration on August 31, 1998, and should be cost-effective in reducing diarrheal diseases in industrialized countries. The vaccine's efficacy and cost effectiveness in developing countries should be evaluated. PMID- 9866731 TI - Cell-to-cell signaling and Pseudomonas aeruginosa infections. AB - Pseudomonas aeruginosa is a bacterium responsible for severe nosocomial infections, life-threatening infections in immunocompromised persons, and chronic infections in cystic fibrosis patients. The bacterium's virulence depends on a large number of cell-associated and extracellular factors. Cell-to-cell signaling systems control the expression and allow a coordinated, cell-density-dependent production of many extracellular virulence factors. We discuss the possible role of cell-to-cell signaling in the pathogenesis of P. aeruginosa infections and present a rationale for targeting cell-to-cell signaling systems in the development of new therapeutic approaches. PMID- 9866733 TI - Chlamydia pneumoniae and cardiovascular disease. AB - Chlamydia pneumoniae is a ubiquitous pathogen that causes acute respiratory disease. The spectrum of C. pneumoniae infection has been extended to atherosclerosis and its clinical manifestations. Seroepidemiologic studies have associated C. pneumoniae antibody with coronary artery disease, myocardial infarction, carotid artery disease, and cerebrovascular disease. The association of C. pneumoniae with atherosclerosis is corroborated by the presence of the organism in atherosclerotic lesions throughout the arterial tree and the near absence of the organism in healthy arterial tissue. C. pneumoniae has also been isolated from coronary and carotid atheromatous plaques. To determine whether chronic infection plays a role in initiation or progression of disease, intervention studies in humans have been initiated, and animal models of C. pneumoniae infection have been developed. This review summarizes the evidence for the association and potential role of C. pneumoniae in cardiovascular disease. PMID- 9866734 TI - Bacterial symbiosis in arthropods and the control of disease transmission. AB - Bacterial symbionts may be used as vehicles for expressing foreign genes in arthropods. Expression of selected genes can render an arthropod incapable of transmitting a second microorganism that is pathogenic for humans and is an alternative approach to the control of arthropod-borne diseases. We discuss the rationale for this alternative approach, its potential applications and limitations, and the regulatory concerns that may arise from its use in interrupting disease transmission in humans and animals. PMID- 9866735 TI - Genetic epidemiology of infectious diseases in humans: design of population-based studies. AB - The spread and clinical manifestations of an infection in human populations depend on a variety of factors, among them host genetics. Familial linkage studies used in genetic epidemiology to identify host genes test for nonrandom segregation of a trait with a few candidate chromosomal regions or any regions in the genome (genomewide search). When a clear major gene model can be inferred and reliable epidemiologic information is collected (e.g., in schistosomiasis), parametric linkage studies are used. When the genetic model cannot be defined (e.g., in leprosy and malaria), nonparametric linkage studies (e.g., sibling-pair studies) are recommended. Once evidence of linkage is obtained, the gene can be identified by polymorphisms strongly associated with the trait. When the tested polymorphism is in strong linkage disequilibrium with the disease allele or is the disease allele itself (e.g., in HIV infection and malaria), association studies can directly identify the disease gene. Finally, the role of the detected polymorphism in causing the trait is validated by functional studies. PMID- 9866737 TI - Mutators and long-term molecular evolution of pathogenic Escherichia coli O157:H7. AB - It has been proposed that an increased mutation rate (indicated by the frequency of hypermutable isolates) has facilitated the emergence of Escherichia coli O157:H7. Analysis of the divergence of 12 genes shows no evidence that the pathogen has undergone an unusually high rate of mutation and molecular evolution. PMID- 9866738 TI - Increasing hospitalization and death possibly due to Clostridium difficile diarrheal disease. AB - This study calculated yearly estimated national hospital discharge (1985 to 1994) and age-adjusted death rates (1980 to 1992) due to bacterial, viral, protozoal, and ill-defined enteric pathogens. Infant and young child hospitalization (but not death) rates in each category increased more than 50% during 1990 to 1994. Age-adjusted death and hospitalization rates due to enteric bacterial infections and hospitalizations due to enteric viral infections have increased since 1988. The increases in hospitalization and death rates from enteric bacterial infections were due to a more than eightfold increase in rates for specified enteric bacterial infections that were uncoded during this period (ICD9 00849). To identify bacterial agents responsible for most of these infections, hospital discharges and outpatient claims (coded with more detail after 1992) were examined for New Mexico's Lovelace Health Systems for 1993 to 1996. Of diseases due to uncoded enteric pathogens, 73% were due to Clostridium difficile infection. Also, 88% of Washington State death certificates (1985 to 1996) coded to unspecified enteric pathogen infections (ICD0084) listed C. difficile infection. PMID- 9866736 TI - Insecticide resistance and vector control. AB - Insecticide resistance has been a problem in all insect groups that serve as vectors of emerging diseases. Although mechanisms by which insecticides become less effective are similar across all vector taxa, each resistance problem is potentially unique and may involve a complex pattern of resistance foci. The main defense against resistance is close surveillance of the susceptibility of vector populations. We describe the mechanisms of insecticide resistance, as well as specific instances of resistance emergence worldwide, and discuss prospects for resistance management and priorities for detection and surveillance. PMID- 9866739 TI - Introduction of Aedes albopictus into a La Crosse virus--enzootic site in Illinois. AB - In late summer and fall 1997, Aedes albopictus mosquitoes were found in Peoria, Illinois, a long recognized focus of La Crosse virus transmission. Larvae were found in tires and other artificial containers, biting adults were recovered, and eggs were collected in oviposition traps within a 25-ha area. One chipmunk trapped < 0.25 km from the infested area tested positive for neutralizing antibodies against La Crosse virus. PMID- 9866740 TI - Mycobacterium canettii, the smooth variant of M. tuberculosis, isolated from a Swiss patient exposed in Africa. AB - An exceptionally smooth and glossy morphotype of Mycobacterium tuberculosis complex was isolated from a 56-year-old Swiss patient with mesenteric tuberculosis. Direct 16S rRNA sequence analysis of the hypervariable signature gene regions revealed a 100% homology to the specific M. tuberculosis complex sequence. Spoligotyping and restriction fragment length polymorphism analyses using the insertion sequences IS6110 and IS1081 and the polymorphic GC-rich sequence as additional genetic markers identified the isolate as the novel taxon M. canettii. Like a Somali child with a similar case, this patient probably contracted the infection in Africa, which raises questions about the geographic distribution of M. canettii. PMID- 9866741 TI - Human infections with Shiga toxin-producing Escherichia coli other than serogroup O157 in Germany. AB - We investigated different types of Shiga toxin-producing Escherichia coli (STEC) not belonging to serogroup O157 for their role as human pathogens. Non-O157 STEC isolated from 89 human patients in Germany were characterized according to serotypes, virulence markers, and association with human illness. EaeA-positive STEC were isolated from 54 (60.7%) of the patients and were frequently associated with severe diarrheal disease, hemolytic uremic syndrome, and young age. EaeA negative STEC were found in 35 (39.3%) of the patients and were more associated with clinically uncomplicated cases and adult patients. For pediatric patients, a serotype-independent diagnosis of STEC is recommended. PMID- 9866742 TI - New Orientia tsutsugamushi strain from scrub typhus in Australia. AB - In a recent case of scrub typhus in Australia, Orientia tsutsugamushi isolated from the patient's blood was tested by sequence analysis of the 16S rDNA gene. The sequence showed a strain of O. tsutsugamushi that was quite different from the classic Karp, Kato, and Gilliam strains. The new strain has been designated Litchfield. PMID- 9866743 TI - Streptococcus parasanguinis: new pathogen associated with asymptomatic mastitis in sheep. AB - We describe two unusual cases in sheep of subclinical mastitis caused by Streptococcus parasanguinis. This bacterium has been associated with the development of experimental endocarditis; its presence at relatively high concentrations in apparently healthy sheep milk may pose a health risk in persons with predisposing heart lesions. PMID- 9866745 TI - Sporadic STEC O157 infection: secondary household transmission in Wales. AB - We conducted a study to quantify and characterize household transmission of Shiga toxin (Vero cytotoxin)-producing Escherichia coli O157 (STEC O157) following sporadic infection in Wales. Through total population surveillance, we identified 83 index case-patients and their household contacts. We screened fecal samples submitted from household contacts for STEC O157 and calculated the household transmission rate for sporadic STEC O157 infection to be 4% to 15%. Household contacts in groups at high risk (particularly children under 5 years of age) present a risk of spreading the infection in the wider community. PMID- 9866744 TI - Introduction of HIV-2 and multiple HIV-1 subtypes to Lebanon. AB - HIV genetic variability, phylogenetic relationships, and transmission dynamics were analyzed in 26 HIV-infected patients from Lebanon. Twenty-five specimens were identified as HIV-1 and one as HIV-2 subtype B. The 25 strains were classified into six env-C2-V3 HIV-1 subtypes: B (n = 10), A (n = 11), C (n = 1), D (n = 1), G (n = 1), and unclassifiable. Potential recombinants combining parts of viral regions from different subtypes Aenv/Dpol/Agag, Genv/Apol, and the unclassifiable-subtype(env)/unclassifiable-subtype(pol)/Agag were found in three patients. Epidemiologic analysis of travel histories and behavioral risks indicated that HIV-1 and HIV-2 subtypes reflected HIV strains prevalent in countries visited by patients or their sex partners. Spread of complex HIV subtype distribution patterns to regions where HIV is not endemic may be more common than previously thought. Blood screening for both HIV-1 and HIV-2 in Lebanon is recommended to protect the blood supply. HIV subtype data provide information for vaccine development. PMID- 9866747 TI - Salmonella enteritidis PT6: another egg-associated salmonellosis? AB - Salmonella Enteritidis phage type 6 (PT6) increased dramatically in the United Kingdom during 1997. The sharp rise suggests that PT6 contamination has spread rapidly throughout a basic food commodity; however, the source and food vehicle remain unknown. We present evidence from three outbreaks suggesting a possible link between PT6 and eggs. Poor documentation of the egg supply network continues to pose problems for public health investigators. Thorough investigation of all future PT6 outbreaks and case-control studies of sporadic infections are needed to confirm the etiology of PT6 infection. PMID- 9866746 TI - Ehrlichia infection in Italy. AB - Immunoglobulin M seroconversion to Ehrlichia chaffeensis was documented in U.S. citizens bitten by ticks in Sardinia. Seven cases of suspected ehrlichiosis in local residents were not confirmed by laboratory tests. In Alpine areas antibodies to E. phagocytophila were detected in persons at high risk, i.e., foresters (8.6%) and hunters (5.5%), and in controls (1.5%). Of 153 persons bitten by ticks, only one was Ehrlichia antibody-positive after 6 months. PMID- 9866748 TI - Reemergence of epidemic malaria in the highlands of western Kenya. AB - Hospital records (1990-1997) of a tea company in the Kericho district, western Kenya, showed malaria epidemics almost annually from May to July, with an annual attack rate of 50%, 857 hospitalizations per 100,000 per year, and 42 deaths per 100,000 per year; 32% of deaths in hospitalized patients were caused by malaria. A questionnaire survey (June 1997) of 244 patients hospitalized for malaria showed that only 8% had traveled to an area with known malaria transmission 30 days before diagnosis. The increasing malaria incidence may be due to drug resistance. PMID- 9866749 TI - Murine typhus in travelers returning from Indonesia. AB - We report the first three documented cases of murine typhus imported into Europe from Indonesia, discuss clues for the diagnosis of the disease, and urge that murine fever be considered in the diagnosis of febrile disease in travelers. PMID- 9866750 TI - Differentiating human from animal isolates of Cryptosporidium parvum. AB - We analyzed 92 Cryptosporidium parvum isolates from humans and animals by a polymerase chain reaction/restriction fragment length polymorphism method based on the thrombospondin-related anonymous protein 2 gene sequence. Used as a molecular marker, this method can differentiate between the two genotypes of C. parvum and elucidate the transmission of infection to humans. PMID- 9866751 TI - An outbreak of hantavirus pulmonary syndrome, Chile, 1997. AB - An outbreak of 25 cases of Andes virus-associated hantavirus pulmonary syndrome (HPS) was recognized in southern Chile from July 1997 through January 1998. In addition to the HPS patients, three persons with mild hantaviral disease and one person with asymptomatic acute infection were identified. Epidemiologic studies suggested person-to-person transmission in two of three family clusters. Ecologic studies showed very high densities of several species of sigmodontine rodents in the area. PMID- 9866752 TI - Host genetics of infectious diseases: old and new approaches converge. PMID- 9866753 TI - Does restricted distribution limit access and coverage of yellow fever vaccine in the United States? PMID- 9866754 TI - Aedes aegypti in Tucson, Arizona. PMID- 9866755 TI - Can the military contribute to global surveillance and control of infectious diseases? PMID- 9866756 TI - Dual infection with Ehrlichia chaffeensis and a spotted fever group rickettsia: a case report. PMID- 9866757 TI - Reemergence of Plasmodium vivax malaria in the Republic of Korea. PMID- 9866758 TI - Hospitalizations after the Persian Gulf War. PMID- 9866759 TI - Atypical squamous cell carcinoma of the larynx and hypopharynx: radiologic features and pathologic correlation. AB - The objective of this study was to analyze the radiologic features of atypical forms of squamous cell cancer and correlate them with clinical, endoscopic, and histopathologic findings. The CT and MRI images of 31 patients with atypical forms of squamous cell carcinoma were reviewed retrospectively and the radiologic findings were correlated with clinical, endoscopic, and histopathologic findings. Histopathologic diagnoses included undifferentiated carcinoma of nasopharyngeal type (n = 8), verrucous carcinoma (n = 18), spindle cell carcinoma (n = 3), and basaloid cell carcinoma (n = 2). Undifferentiated carcinoma of nasopharyngeal type was located in the supraglottis or piriform sinus beneath an intact mucosa and initial endoscopic biopsy was most often negative. The discrepancy between an intact mucosa at endoscopy and a solid mass with homogenous enhancement at CT or MRI was characteristic for these tumors and warranted further investigations to obtain the definitive histologic diagnosis. Verrucous carcinoma displayed characteristic clinical, radiologic, and pathologic features, namely, an exophytic tumor arising from the glottic level displaying a rugged surface with finger-like projections but with only minor submucosal infiltration. Spindle cell carcinoma appeared as a polypoid mass with a thin stalk arising from the supraglottis. Basaloid cell carcinoma displayed a distinct lobulated enhancement pattern which was observed on contrast-enhanced T1-weighted SE images. Although the MR and CT features of atypical forms of squamous cell carcinoma cannot be considered pathognomonic they should raise the differential diagnosis even if endoscopic biopsy has been negative. The radiologist's awareness of the appearance of these unusual tumors on CT and MR images may greatly facilitate the diagnostic work-up and helps to guide the endoscopist to the adequate biopsy site in order to establish the correct diagnosis. PMID- 9866760 TI - Assessment of tumour invasion into the mandible: the value of different imaging techniques. AB - In oral carcinomas close to the mandible, tumour invasion of the mandible is important in selecting segmental or marginal resection. Imaging may play a role in assessing tumour invasion. This study compares the accuracy of panoramic X ray, CT and MR imaging in assessing invasion of the mandible in 29 patients. At histopathology, 6 patients had mandible erosion, 12 had invasion and 11 had an intact mandible. Magnetic resonance imaging had the highest sensitivity (94%), but a low specificity (73%), with 3 of 11 intact mandibles interpreted as positive. Furthermore, MR often overestimated the extent of tumour invasion. On the other hand, CT and panoramic X-ray had a lower sensitivity (64 and 63%, respectively) and a higher specificity (89 and 90%, respectively). Computed tomography (using 5-mm sections) and panoramic X-ray had a similar accuracy, and negative findings do not exclude invasion. Magnetic resonance imaging was the most sensitive technique but had more false positives and frequently overestimated the extent of tumour invasion. Because none of the radiological techniques are accurate enough, clinical examination seems at present to remain the most important modality in deciding between segmental and marginal resection. Tumour invasion at CT or panoramic X-ray is a strong argument for a segmental resection. PMID- 9866761 TI - A new volumetric CT machine for dental imaging based on the cone-beam technique: preliminary results. AB - The objective of this paper is to present a new type of volumetric CT which uses the cone-beam technique instead of traditional fan-beam technique. The machine is dedicated to the dento-maxillo-facial imaging, particularly for planning in the field of implantology. The main characteristics of the unit are presented with reference to the technical parameters as well as the software performance. Images obtained are reported as various 2D sections of a volume reconstruction. Also, measurements of the geometric accuracy and the radiation dose absorbed by the patient are obtained using specific phantoms. Absorbed dose is compared with that given off by spiral CT. Geometric accuracy, evaluated with reference to various reconstruction modalities and different spatial orientations, is 0.8-1% for width measurements and 2.2% for height measurements. Radiation dose absorbed during the scan shows different profiles in central and peripheral axes. As regards the maximum value of the central profile, dose from the new unit is approximately one sixth that of traditional spiral CT. The new system appears to be very promising in dentomaxillo-facial imaging and, due to the good ratio between performance and low cost, together with low radiation dose, very interesting in view of large scale use of the CT technique in such diagnostic applications. PMID- 9866762 TI - Percutaneous ethanol injection into parathyroid adenomas: mid- and long-term results. AB - The aim of this study was to evaluate mid- and long-term results of ultrasonically guided ethanol injection into parathyroid adenomas. From 1988 to 1996, 27 patients (mean age 77 +/- 13 years) were treated for parathyroid adenomas by percutaneous ethanol injection. The survey included clinical information, plasma calcium, phosphorus and PTH (1-84) evaluation. Follow-up lasted 22.6 +/- 10 months. No major complications were observed for 63 ethanol injections. Biochemical recovery was 58%, biochemical improvement at 3 months was 33%, and failure was 7%. Four of 15 cured patients presented a recurrence of the disease 1 or 2 years after the first treatment. Ultrasonically guided ethanol injection can be useful in the treatment of parathyroid adenomas when surgery is not possible. The immediate results are interesting but not as good as those obtained with surgery. A regular biochemical survey is necessary so that recurrences can be recognized and treated at an early stage. PMID- 9866763 TI - CT findings of hydatid cyst with unusual location: infratemporal fossa. AB - Hydatid disease has a high incidence in the countries of the temperate zones such as Turkey. Only few cases in the head and neck region have been reported in the literature. Our case, an unusual localization of hydatidosis, i.e. hydatid disease of the infratemporal fossa of a 9-year-old male patient suffering from a swelling of the left maxillary region which was diagnosed by CT, is presented. The lesion visualized on CT images was compressing the neighbouring structures. The possible diagnosis was made based on the images obtained from CT examination. PMID- 9866764 TI - Functional MRI of the brain: localisation of eloquent cortex in focal brain lesion therapy. AB - The aim of this study was to assess the feasibility of functional MRI (fMRI) in a clinical environment on a large patient group, and to evaluate the pretherapeutic value of localisation of eloquent cortex. Forty patients with focal brain lesions of different origin were studied using fMRI. Functional information was obtained using motor, somatosensory, auditory and phonological stimuli depending on the localisation of the lesions. To obtain information about the spatial accuracy of fMRI, the results were compared with postoperative electrocortical stimulation. Two patients with secondary trigeminal neuralgia were scanned using a motor protocol and were implanted with an extradural plate electrode. Imaging was successful in 40 of 42 patients (including the 2 with trigeminal neuralgia). These patients were analysed for strength of activation, the relation of the lesion to activation sites and the presence of mass effect. The correlation between these data and surgical findings provided significant additional clinical information. Functional MRI can be accurately performed in patients with focal brain lesions using a dedicated approach. Functional MRI offers important clinical information as a contribution to a decrease in posttherapeutic morbidity. The accuracy of the technique can be confirmed by other modalities, including invasive cortical electrostimulation. PMID- 9866765 TI - Dimensions of the lumbar spinal canal: variations and correlations with somatometric parameters using CT. AB - The aim of this study was to investigate the correlation of vertebral dimensions with somatometric parameters in patients without clinical symptoms and radiological signs of central lumbar spinal stenosis. One hundred patients presenting with low back pain or sciatica were studied with CT. In each of the L3, L4 and L5 vertebra three slices were taken with the following measurements: 1. Slice through the intervertebral disc: (a) spinal canal area; (b) interarticular diameter; (c) interligamentous diameter. 2. Slice below the vertebral arcus: (a) dural sac area; (b) vertebral body area. 3. Pediculolaminar level: (a) anteroposterior diameter and interpedicular diameter of the spinal canal; (b) spinal canal area; (c) width of the lateral recesses. The Jones Thomson index was also estimated. The results of the present study showed that there is a statistically significant correlation of height, weight and age with various vertebral indices. The conventional, widely accepted, anteroposterior diameter of 11.5 mm of the lumbar spinal canal is independent of somatometric parameters, and it is the only constant measurement for the estimation of lumbar spinal stenosis with a single value. The present study suggests that there are variations of the dimensions of the lumbar spinal canal and correlations with height, weight and age of the patient. PMID- 9866766 TI - Somatosensory evoked potentials: a simple neurophysiological monitoring technique in supra-aortal balloon test occlusions. AB - The aim of this study was to evaluate somatosensory evoked potentials (SEP) as a simple neurophysiological monitoring method in supra-aortal balloon test occlusions (BTO). The 13 patients examined had carcinoma of the larynx (n = 6), nasopharyngeal malignancy (n = 3), sphenoid meningioma (n = 2), mycotic aneurysm (n = 1), and a tumor of the carotid body (n = 1). Transfemoral balloon occlusion of the internal carotid artery was maintained for 30 min and monitored both clinically and by SEP. None of the 13 patients showed neurological deficits or pathological changes in the SEP. No thromboembolic complications occurred during BTO. In 11 cases the cervical vessels could be spared or reconstructed during surgery. No neurological symptoms occurred during the clinical follow-up. One patient died the day after BTO due to cardiac arrest. Neurophysiological monitoring of preoperative BTO using SEP is a technically simple method. A similar method has been shown to have a high sensitivity and reliability in carotid surgery. PMID- 9866767 TI - MRI findings in cerebral fat embolism. AB - The purpose of this retrospective study was to demonstrate the MRI features of cerebral manifestations in patients with fat embolism syndromes in comparison with cerebral CT (CCT). Magnetic resonance imaging was performed according to standard protocols revealing multiple small non-confluent hyperintense intracerebral lesions larger than 2 mm on proton-density and T2-weighted images to various extents in three of four patients with clinically suspected cerebral fat embolism. Cerebral CT was negative in all patients. Our findings confirm that MRI can detect cerebral fat embolism with a higher sensitivity than CCT. Thus, MRI should be the first choice for imaging of cerebral fat embolism. PMID- 9866768 TI - Parasellar dermoid tumor with intra-tumoral hemorrhage. AB - We report a case of parasellar dermoid tumor with intra-tumoral hemorrhage. It is rare for a dermoid tumor that hemorrhage was detected as high attenuation on the initial CT. In the present case, the tumor content included a little fat component and mostly cholesterin-rich fluid which resulted in extremely low signal intensity on T2-weighted and high signal on T1-weighted MR images. In addition to this, hemosiderin accumulation in the tumor could be the reason for low signal intensity on T2-weighted images. PMID- 9866769 TI - Primary tumours of the greater omentum. AB - Primary tumours of the greater omentum are very rare. Ultrasound allows the detection and characterization of such lesions, but determination of their precise anatomical location is usually difficult by US. Computed tomography determines the omental origin of the tumour. Thus, when US reveals an abdominal tumour of unknown origin, the possibility of an omental tumour, although rare, must be kept in mind and CT should be performed. PMID- 9866770 TI - Causes, clinical features and non-operative management of bile leaks. AB - This paper evaluates the management of 31 patients with bile leaks identified over a 7-year period. Leaks complicated cholecystectomy in 19 patients (11 laparoscopic, 8 open), interventional procedures in 10 (including surgery in 1), trauma in one and was spontaneous in one case. Confirmation of the diagnosis typically lagged behind the onset of symptoms (mean for the group 4.2 days), indicating that a high index of suspicion is required in at-risk patients with typical symptoms. These include abdominal pain or distension, fever, bile leaking along a drain, jaundice, abnormal liver function tests and elevated white cell count. Two post-surgical bile leaks required surgical drainage of abdominal cellections. The remainder were successfully managed by non-operative methods including percutaneous drainage, endoscopic retrograde cholangiography with or without sphincterotomy or stent placement and percutaneous stenting. The spontaneous leak and all bile leaks complicating interventional procedures were managed non-operatively, although six patients in this group died due to the underlying malignant pathology. Only the patient with self-inflicted transection of the bile duct died directly from the complications of the bile leak. Although this is a varied, small series, we conclude that the majority of bile leaks can be managed by non-operative techniques. Whilst endoscopy is the primary modality for treatment, percutaneous techniques are crucially important for the management of complex cases and endoscopic failure. PMID- 9866771 TI - Optimizing liver contrast in helical liver CT: value of a real-time bolus triggering technique. AB - Optimal liver enhancement during portal venous-phase helical CT is crucial in the detection of parenchymal liver lesions. In the prospective study reported here we investigated the effects of a real-time bolus-tracking system on mean and maximal liver enhancement. In 79 patients referred to us for abdominal CT we injected 120 ml of non-ionic contrast (300 mg I/ml) at a rate of 3 ml/s. After a nonintravenous contrast upper abdominal scan a portal venous phase was performed. In 39 patients (mean weight 72.6 +/- 18.7 kg, range 48-139 kg) real-time bolus tracking was performed using the CARE Bolus software (Siemens, Erlangen, Germany). The software performs repetitive low-dose test scans in a preselected region of interest and measures the Hounsfield attenuation and liver enhancement in real-time. After a critical threshold (we selected 31 HU) is surpassed, the software starts diagnostic spiral scanning. Our control consisted of 40 patients weighing 51-100 kg (mean 73.2 +/- 11.1 kg) who were scanned with a fixed, preselected start delay of 80 s. Mean hepatic enhancement was 54.0 +/- 9.9 HU (range 33.3-74 HU) in 37 automatically triggered patients, mean peak hepatic enhancement 64.6 +/- 12.6 HU (range 42.0-91.8 HU). In 2 patients of the study group scanning had to be started manually. In the control group with fixed delay mean enhancement was 48.3 +/- 9.2 HU (range 33.8-71.6 HU) and peak enhancement 55.5 +/- 9.7 HU (range 39.7-81.0 HU). Differences were significant (p < 0.05, Student's t-test). Real-time bolus tracking significantly increased mean hepatic enhancement and may improve portal venous hepatic CT scanning. PMID- 9866772 TI - Development of extrahepatic arterial blood supply to the liver during hepatic arterial infusion chemotherapy. AB - The aim of this study was to evaluate the correlation of development of the collateral circulation to the liver during hepatic arterial infusion chemotherapy (HAIC) with the presence of hepatic tumours adjacent to the hepatic surface, and with pretreatment occlusion of aberrant hepatic arteries. In 102 patients with unresectable malignant hepatic tumours treated with HAIC using an implantable port system, development of collaterals to the liver was assessed with CT arteriography using the implantable port and pre- and postoperative angiography. Aberrant hepatic arteries, if present, were occluded prior to treatment for hepatic arterial redistribution. Collaterals to the liver were seen in 29 patients, who had 35 areas with collateral perfusion: 22 areas were in the right posterosuperior area, 6 in the left peripheral area and 7 in the right or left lobar area. Collaterals were revealed more frequently in patients with hepatic tumours adjacent to the hepatic surface than in those without hepatic tumours in peripheral areas in the liver (p < 0.0001). In addition, collaterals developed more frequently in patients with an aberrant hepatic arterial anatomy compared with those with conventional anatomy (p = 0.0007). Our results indicated that patients with hepatic tumours adjacent to the hepatic surface and with pretreatment occlusion of aberrant hepatic arteries had the potential to develop collaterals to the liver during HAIC. PMID- 9866773 TI - Diagnosis and nonsurgical management of bile leak complicated by biloma after blunt liver injury: report of two cases. AB - We report on two patients with biliary tract injury and associated biloma following blunt abdominal trauma. Both patients underwent emergency surgery because of hemodynamic instability and bloody peritoneal aspiration. Computed tomography in the postoperative days showed severe hepatic parenchymal injury and the presence of hypodense collections with intraparenchymal and subcapsular extension, suggestive for biloma, but otherwise failed to demonstrate the exact location of the bile duct injury. One of them underwent temporary percutaneous drainage. Bile duct injury was well demonstrated on endoscopic retrograde cholangiography (ERCP) and treated by endobiliary stent placement. This report advocates the use of ERCP and endobiliary stenting in the management of biliary injury resulting from liver trauma. PMID- 9866774 TI - MRI, CT, US and ERCP in the evaluation of bile duct hamartomas (von Meyenburg complex): a case report. AB - A case of multiple bile duct hamartomas (von Meyenburg complex) of the liver accompanied by exudative and in part necrotizing pancreatitis is presented. Magnetic resonance imaging (fat suppressed, T2-weighted images with prolonged echo time) could exclude diffuse tumor infiltration of the liver, which had not been possible with CT, sonography, or ERCP. To our knowledge, no comparable case has been reported. PMID- 9866775 TI - Solitary true cyst of the pancreas. AB - We report a case of a woman diagnosed with a solitary true cyst of the pancreas. CT and especially MRI guided us in the diagnosis of this benign lesion. The cyst has been surgically removed because of secondary bile-duct obstruction causing painless jaundice. Clinical history, laboratory, imaging, and histological findings are reported. PMID- 9866776 TI - Standardized CT examination of the multitraumatized patient. AB - The aim of this study was to develop a standardized non-helical-CT protocol including head, body and proximal extremities in order to achieve a good time efficiency and diagnostic accuracy in the initial radiological evaluation of the multitraumatized patient. A total of 111 circulatory stable blunt trauma patients, brought in to a trauma level II-III hospital, were examined according to a standardized CT protocol. After examining the head with contiguous 10-mm slices without i.v. contrast medium injection, the trunk was examined with 10-mm slices every 30 mm through thorax-abdomen-pelvis with i.v. contrast medium enhancement (occasionally modified). All data in the medical reports were collected and used as "end-point", and the outcome of the CT examination was compared with this final diagnosis. Mean examination time was 20 min (range 12-32 min). In total, 55 head injuries, 89 thoracic injuries, 27 abdominal/pelvic injuries and 62 fractures were found. Computed tomography correctly identified the injuries, except one brain stem injury, one contusion/rupture of the heart, one hepatic injury, two intestinal injuries, eight vertebral injuries and one joint dislocation. A standardized non-helical-CT examination of the head and body may be achieved in 20 min. Its diagnostic accuracy was high, except for vertebral column injuries, which is why we recommend it as the method of choice for initial radiological examination of multitraumatized patients. When available, helical scanning would improve both examination speed and accuracy. PMID- 9866777 TI - The anterior tilt of the acromion: radiographic evaluation and correlation with shoulder diseases. AB - The aim of this study was to test whether the anterior tilt of the acromion can be objectively evaluated on lateral radiographs, and whether there is a relation between this anatomical feature and the most common shoulder diseases. Lateral radiographs of 15 dried scapulas were performed in neutral position and with 5 degrees of caudal, cranial, anterior and posterior angulations. Two hundred and forty-three shoulders, both asymptomatic and affected by chronic and post traumatic impingement, calcific tendinitis and instability, were examined by conventional radiography. The presence of rotator-cuff tears was investigated by sonography. A method was elaborated to obtain reproducible lateral radiographs and to determine the acromial tilt angle. Inter- and intraobserver and inter- and intraoperator variations in measurements were evaluated. Variations in tube angulation produced changes in tilt-angle values on dried scapulas. The average tilt angle for the chronic impingement and the instability groups were significantly different from the post-traumatic, calcific tendinitis and control groups. Rotator-cuff tears were significantly more frequent in the chronic impingement group and related to a more acute tilt angle. The inter- and intraobserver variability coefficients were 0.95 and 0.98, whereas the inter- and intraoperator variability coefficients were 0.94 and 0.96, respectively. Conventional radiography using fluoroscopy for positioning is a well-reproducible method for the evaluation of acromial tilt. There is a significant difference in tilt-angle values between some groups of subjects examined, but the lack of specificity limits the clinical importance of such measurement. PMID- 9866778 TI - Gorham disease: correlation of MR findings with histopathologic changes. AB - Gorham disease is a rare disorder of unknown etiology characterized by bone destruction and abnormal proliferation of thin-walled vascular channels including lymphatic capillaries. Starting monocentrically in a single bone, the angiomatous masses in this disease extend to adjacent bones and soft tissues without respecting articular barriers. Herein we report a case of Gorham disease with its MR and histopathologic appearance. PMID- 9866779 TI - MR imaging of muscular sarcoidosis after steroid therapy. AB - The nodular type of muscular sarcoidosis has been known to show characteristic MR findings: however, MR imaging features after steroid therapy have not been reported. A 48-year-old man with nodular type of muscular sarcoidosis is reported. Prior to steroid therapy, axial MR images showed peripheral increased signal intensity and central star-shaped decreased signal intensity. Coronal images showed an inner stripe of decreased signal intensity and outer stripes of increased signal intensity. After steroid therapy, axial images showed only the central star-shaped area of decreased signal intensity. Coronal images showed only the inner stripe of decreased signal intensity. It is important to know that the central area will continue to exist after steroid therapy. PMID- 9866781 TI - Non-communicating intramuscular ganglia. AB - Intramuscular ganglia are rare. Most of the previously reported cases were connected with an adjacent joint. We present the imaging findings in three patients who had intramuscular ganglia that were not connected with a joint. Magnetic resonance showed a septated, encapsulated mass that was isoor hypointense to muscle on T1-weighted and hyperintense on T2-weighted images. A post-contrast T1-weighted scan in one patient showed minimal capsular enhancement. Ultrasound performed in one case showed an encapsulated, anechoic mass. PMID- 9866780 TI - Soft tissue Burkitt's lymphoma: radiological findings. AB - An unusual case is reported of a soft tissue mass in the lower extremity, without bone involvement, in an 85-year-old woman; the histopathological diagnosis was Burkitt's lymphoma. Pertinent clinical history, histological examination, and imaging procedures allowed early diagnosis. To our knowledge, the radiological findings in Burkitt's lymphoma with this unusual clinical presentation have not been described previously. PMID- 9866782 TI - Enhancement of intramammary lymph nodes with lymphoid hyperplasia: a potential pitfall in breast MRI. AB - We present three cases of breast lesions labeled as probable intramammary lymph nodes that showed an increase in size on follow-up mammography. Contrast-enhanced MRI was performed and the three lesions showed strong and rapid uptake of the intravenous contrast. Core needle biopsy established the diagnosis of lymphoid hyperplasia in all three patients. Because intramammary lymph nodes affected by benign processes can present findings similar to malignant lesions, the usefulness of contrast-enhanced MRI in these cases is controversial. PMID- 9866783 TI - Mucinous breast carcinoma showing as a cluster of suspicious microcalcifications on mammography. AB - We report a case of mucinous breast carcinoma whose mammographic features consisted of a suspicious cluster of microcalcifications as the unique sign of malignancy. The lesion was non-palpable and measured approximately 2 cm at the greatest diameter. The microcalcifications showed several morphologies: round, pleomorphic, and fine shapes were identified. The total number of microcalcifications was > 30 and the number per square centimeter varied from 10 to 20. The histological calcifications showed good correlation with the mammographic ones and were localized predominantly at the periphery of the tumor inside ducts with ductal carcinoma in situ or in the acellular mucin. Two types were observed: psammomatous and gross-irregular calcifications. To our knowledge, only one case with similar findings has been reported previously. PMID- 9866784 TI - High-resolution computed tomographic appearance of MALToma of the lung. AB - MALToma of the lung is rare and advances in molecular techniques have only recently allowed accurate diagnostic classification of the previously termed "pseudolymphomas" by demonstrating that many are monoclonal B-cell proliferations of MALT tissue and therefore true low-grade lymphomas. No significant previous contribution was found in the literature regarding the high-resolution CT appearance (HRCT) of these tumours. We describe the high-resolution CT appearances in five cases presenting to our institution from 1994 to 1997. The HRCT scans (1-mm sections at 10- to 15-mm intervals) were performed as the opacities seen radiographically were thought to be part of a diffuse lung process. In one patient a spiral sequence was performed through the main airway. Multifocal, ill-defined nodules containing air bronchograms were seen in four cases and focal lobar consolidation in one case. Interlobular septal thickening, centrilobular micronodules and bronchial wall thickening were seen in two cases. Mediastinal lymphadenopathy and pleural reaction do not appear to be characteristic features. The appearance of multifocal consolidation is similar to that seen in bronchoalveolar cell carcinoma and cryptogenic organising pneumonia. PMID- 9866785 TI - Respiratory bronchiolitis: radiographic and CT findings in a pathologically proven case. AB - A small number of cases of cigarette-smoking-associated respiratory bronchiolitis (RB) with positive findings on the chest radiograph have been reported in the literature. High-resolution computed tomography (HRCT) findings are available in even fewer cases. We describe the case of an asymptomatic female smoker presenting with a reticulomicronodular infiltrate on a routine chest radiograph. High-resolution CT was characterized by ground-glass opacities and centrilobular micronodules with an upper lobe predominance. Surgical biopsy revealed peribronchiolar lesions, with accumulation of brown pigmented macrophages in the lumen of alveolar and bronchiolar lumen, consistent with the pathologic diagnosis of RB. PMID- 9866786 TI - CT and MRI appearances of a thoracic chordoma. AB - A case of a chordoma in the thoracic spine is presented. This is a very rare tumour in this location and should be considered in the differential diagnosis of any posterior mediastinal mass. The appearances on CT and MRI were similar to chordomas described in other locations. On T2-weighted images septae of low signal intensity radiated throughout the large high-signal mass. This feature may be of use in differentiating chordomas from other posterior mediastinal masses. PMID- 9866787 TI - Pulmonary alveolar proteinosis in a child: HRCT findings before and after bronchoalveolar lavage. AB - We demonstrate the high-resolution computed tomography (HRCT) findings of pulmonary alveolar proteinosis in a child before and after bronchoalveolar lavage. The CT pattern in our case differs from the pattern described in previous reports. We found a more homogeneous distribution of the pulmonary changes and a "crazy paving" pattern. High-resolution CT may be helpful in the differential diagnosis of this rare disease and in the follow-up of the pulmonary changes after bronchoalveolar lavage. PMID- 9866788 TI - Mechanical thrombectomy of massive pulmonary embolism using an Arrow-Trerotola percutaneous thrombolytic device. AB - Mechanical thrombectomy of a large central thrombus in massive pulmonary embolism is a new option for the treatment of this serious condition. The special mechanical devices designed to fragmentize a blood clot include the Arrow Trerotola percutaneous thrombolytic device (PTD), the use of which in the pulmonary arteries has not yet been reported. The case of massive embolism into the left pulmonary artery with subsequent collapse is to demonstrate the immediately clinically successful treatment using the PTD. Our initial experience with the PTD shows that its use is a safe and quick procedure. PMID- 9866789 TI - Lymphoproliferative disorders after renal transplantation: role of medical imaging. AB - Post-transplantation lymphoproliferative disorders (PTLD) are a complication of immunosuppressed transplant recipients, and their incidence is reported to be 20 120 times greater than the rate in the general population. After kidney transplantation, PTLD more likely arise within the renal transplant fossa. Radiological patterns of these forms are presented and discussed, according to a review of the literature, and illustrated by cases from our institution. Ultrasound plays an essential role in the early diagnosis of PTLD by detecting a urinary obstruction associated with adenopathy or an ill-defined mass not previously seen. However, in the case of an inconclusive US examination, CT or MRI should be performed to confirm the presence of a mass. Both techniques are useful in evaluating the extension of the process within the transplantation fossa; MRI seems more accurate and can be used for the follow-up, especially after reduction in immunosuppressive therapy without transplant removal. PMID- 9866790 TI - Bilateral renal cell carcinoma in a horseshoe kidney: preoperative assessment with MRI and digital subtraction angiography. AB - Renal cell carcinoma in a horseshoe kidney is an unusual entity. To our knowledge, only 123 cases have been published to date. We report the first bilateral case of two clear-cell carcinomas in an asymmetrically fused kidney. Optimum preservation of renal function after radical tumor removal requires accurate preoperative imaging. Since the vascular supply in fusion anomalies is extremely variable, angiography is mandatory. Magnetic resonance imaging was most suitable to predict the tumor extent and localization, because it simultaneously gave the most comprehensive anatomical overview of the malformation. PMID- 9866791 TI - Nuclear medicine studies of the heart. AB - Nuclear imaging techniques are well established diagnostic tools in clinical cardiology, providing noninvasive information about myocardial perfusion, function and metabolism. The cost-effectiveness of radionuclide imaging in the diagnostic work-up of patients with coronary artery disease has been demonstrated. Additionally, the documented prognostic value of scintigraphic parameters is of clinical importance to guide decision making. Advances in technology, new radiotracers and new applications contribute to continuous growth in the field of nuclear cardiology. Multi-headed gamma camera systems lead to higher spatial resolution and sensitivity of cardiac single photon emission tomography (SPECT), and they also provide the opportunity for attenuation correction or electrocardiographic gating of SPECT images. Objective quantitative values of perfusion, function and metabolism are derived from scintigraphic data by use of improved software and hardware. With the latest developments in tracer technology, imaging of myocardial necrosis, receptor systems and autonomic innervation has become a reality and will lead to new clinical applications in the future. PMID- 9866792 TI - Integrating HIS-RIS-PACS: the Freiburg experience. AB - With the integration of different formerly isolated information systems, such as Hospital Information System (HIS). Radiology Information System (RIS), Picture Archiving and Communication System (PACS) and modalities, we evolve towards an architecture of distributed information processing in radiology, the data being stored in distributed databases. The frontier between the cooperative information systems becomes more subtle than in the past, and it seems more convenient to distinguish the different computer functionalities or services used in radiology than to detail the functions of RIS and PACS. This paper opens with a description of the different computer services used in radiology and how they relate one to another. It also shows how these functionalities could be integrated within a global Radiology Information and Archiving System. Finally, it shows the degree of systems interconnection that can be achieved presently, using the PACS-RIS installation at the University Hospital of Freiburg as an example. PMID- 9866793 TI - Is image selection a useful strategy to decrease the transmission time in teleradiology? A study using 100 emergency cranial CTs. AB - This study examines the suitability of working with a selection of images in a teleradiology consulting system in neurological or neurosurgical emergency situations. The teleradiology system was based on IBM-compatible personal computers, video digitization for data acquisition, and data transmission by Integrated System Digital Network. Forty normal and 60 abnormal emergency cranial computed tomograms were shown to a radiologist on call who presented all cases he regarded as pathologic to a neuroradiologic expert by teleradiology. To reduce transmission time, only a selection of images from the CT study was presented (up to four images per case). For each case the on-call radiologist's diagnosis (D(on call)), the expert's diagnosis on the teleradiology screen (D(monitor)), and the expert's diagnosis on the original film (D(original)) was documented, together with an estimation of the agreement between those diagnoses. There was clinically relevant disagreement between the on-call radiologist's diagnosis and the neuroradiologist's diagnosis based on the image selection on the teleradiology monitor in 23% of cases. A clinically important discrepancy between the neuroradiologist's diagnosis based on the image selection and his diagnosis using the original films was found in 30% of cases. This was due to the presence of clinically relevant information on images not transferred by the on-call radiologist. Image quality of the transferred images was sufficient in all cases. Drastic selection of images from a complete CT study leads to a high rate of incorrect diagnoses and is not appropriate to reduce transmission time in teleradiology. PMID- 9866794 TI - Magnetic resonance imaging of the subarachnoid space. PMID- 9866795 TI - [The classification of cardiomyopathies]. PMID- 9866796 TI - [The role of free radicals in coronary atherosclerosis]. PMID- 9866797 TI - Normal values for left ventricular volumes obtained using gated PET. AB - BACKGROUND: Accurate assessment of left ventricular (LV) volumes provides important insights into myocardial function, being particularly important for diseases of the heart in which there is progressive dilatation of the LV, such as remodeling after myocardial infarction. We have recently validated a method for measuring LV volumes using gated positron emission tomography (PET) with oxygen 15 labeled carbon monoxide (C15O). The present study was performed to establish normal values for LV volumes, in absolute units, using this technique. METHODS: Forty subjects [21 males and 19 females; age 49 +/- 13 years (mean +/- SD), range 24-80; body surface area (BSA) range 1.50-2.18 m2] were studied. End diastolic (EDV) and end systolic volumes (ESV) were calculated by determining the edge of the end diastolic and end systolic blood pool images (obtained using PET with C15O) and summing the volume of the voxels comprising the blood pool. RESULTS: For the overall study population, EDV was 109 +/- 20 ml and ESV 37 +/- 12 ml. When stratified according to gender, these values were: in males, EDV was 117 +/- 18 ml and ESV 42 +/- 18 ml; in females, EDV was 100 +/- 19 ml and ESV 32 +/- 8 ml. The differences between gender groups (EDV: p = 0.0027, ESV: p = 0.0029) were eliminated when volumes were corrected for BSA. CONCLUSIONS: These data will form a database of normal values for comparison with pathological conditions of the heart. PET quantification of LV volumes offers a means of measurement of function using an imaging modality that can also assess regional myocardial perfusion and metabolism during the same study session. PMID- 9866798 TI - Early assessment of viable myocardium after acute myocardial infarction by low dose echo-dobutamine. AB - BACKGROUND: The aim of the study was to evaluate the usefulness of low-dose dobutamine echocardiographic testing performed within 48 hours from anterior AMI in order to identify the extent of viable myocardium and predict its functional outcome. The early echo-dobutamine test was also compared with a predischarge test in order to evaluate the effects of different timing on the accuracy of the test. METHODS: Nineteen consecutive patients, aged 54 +/- 11 years, with a first anterior AMI entered the study. All patients underwent a low-dose dobutamine echocardiographic test within 48 hours from hospital admission and at predischarge. In all the patients, a rest follow-up echocardiogram was performed three months after hospital discharge. Eleven patients underwent a revascularization procedure (7 underwent PTCA and 4 CABG). RESULTS: Of the 159 dyssynergic segments, 26% improved spontaneously at predischarge and 51% improved at the three-month follow-up. Of the 145 predischarge dyssynergic segments, 38% improved at three months. Considering the results on a segmental basis, early low dose dobutamine echocardiography showed a sensitivity of 52%, a specificity of 87%, a positive predictive value of 81%, a negative predictive value of 64% and a diagnostic accuracy of 69% for wall-motion improvement at three months. The predischarge test showed very similar values. A slight enhancement of the sensitivity of both tests was observed considering the akinetic segments only. Finally, considering the amount of segmental reversible dysfunction inside the infarct area in the single patients, early low-dose dobutamine echocardiography showed a sensitivity of 86% and a specificity of 80%. CONCLUSIONS: Our results indicate that: 1) recovery of regional wall motion after AMI is slow and progressive, with substantial improvement ensuing within the first days after infarction; 2) considering results on a segmental basis, low-dose dobutamine echocardiography performed within 48 hours of AMI shows a high specificity but a low sensitivity for late recovery of regional function, although it gave information similar to what was obtained performing the test at predischarge; 3) the efficiency of test can be improved by considering the amount of reversible segmental dysfunction inside the infarct area in the single patients. PMID- 9866799 TI - [Minimally invasive surgery with the Port-Access method. Preliminary experience]. AB - METHODS: Data from the initial experience of 40 patients operated on with the Port-Access technique are reported. Indication to surgery was mitral disease in 24 patients and coronary stenosis in 16 patients. Mean age was 52 years (range 32 75). Operations performed were: 8 mitral valvuloplasties, 16 valve replacements, 9 single CABG (associated with an MVR in one case), 1 double CABG, 6 triple CABG and one quadruple CABG. Coronary endarterectomy was performed in 5 patients and left atrial isolation was associated with MV surgery in 5 cases. RESULTS: There were no operative deaths and every patient was discharged after a mean postoperative stay of 5.5 days (range 3-30). Postoperative course was complicated in 7 patients: surgical revision was necessary in 4 patients due to bleeding (through the mini-thoracotomy incision in 3 cases), 1 pacemaker was implanted for A-V block, one retained pulmonary catheter was removed through the mini thoracotomy without the aid of cardiopulmonary bypass and in one case, there was an emergency conversion to median sternotomy due to a ventricular fibrillation unresponsive to usual resuscitative maneuvers a few hours after surgery. Some of these complications can be ascribed to the learning phase of this new technique and should disappear as experience is increased. CONCLUSIONS: Port-Access surgery is a new minimally invasive technique that utilizes a cardiopulmonary bypass with femoral access and a specialized catheter system that provides endoaortic clamping, pulmonary artery venting and myocardial preservation with infusion of cardioplegic solution in the aortic bulb or in the coronary sinus. Major contraindications to this technique are iliac-femoral disease or severe dilatation of ascending aorta. The aim of the Port-Access technique is to combine the aesthetic and functional advantages of the minimally invasive surgery with the wide range of surgical options that cardiopulmonary bypass can afford (to operate on atrioventricular valves and perform all the CABG that the patient need), without the limitations characteristic of the classic minimally invasive direct coronary artery bypass (MIDCAB) technique. PMID- 9866800 TI - [The use of bilateral mammary artery in myocardial revascularization. The risk factors emergent from a multivariate analysis conducted on 474 patients]. AB - BACKGROUND: The internal mammary artery is used as coronary artery graft conduit because of its superior patency. According to some authors, the bilateral IMA can increase perioperative morbidity. The aim of this study was to determine the risk factors increasing perioperative mortality and morbidity in the use of bilateral IMA. METHODS: We analyzed the data of 474 patients operated consecutively with the use of bilateral IMA between January 1987 and December 1995 at the Department of Cardiac Surgery of the Varese Hospital. The univariate analysis was done on 17 ordinal variables using a "Fisher exact test" and on 4 continuous variables by "pooled-variance t-test" to investigate risk factors for mortality, mediastinitis, superficial wound infection and aseptic dehiscence of the sternum; a p-value lower than 0.1 was used as cut-off point to introduce the variables into a stepwise multiple logistic regression analysis. RESULTS: From the univariate analysis are: postoperative low-output syndrome (p = 0.01), LVEF (p = 0.02) and number of grafts (p = 0.04) are correlated to hospital mortality (1.5%); obesity (p < 0.001) and peripheral arteriopathy (p = 0.009) are correlated to postoperative mediastinitis (5%); obesity (p < 0.001), peripheral arteriopathy (p = 0.009), surgeon (p = 0.001), year of operation (p < 0.001), reoperation for bleeding (p = 0.004) and length of extracorporeal circulation (p = 0.02) are correlated to superficial wound infection (7%); obesity (p = 0.002) and COPD (p = 0.05) are correlated to aseptic dehiscence of the sternum (2%). The multivariate analysis identified low LVEF as the only independent risk factor for hospital mortality (p = 0.03), whereas obesity (p = 0.01) and peripheral vasculopathy (p = 0.03) proved to be correlated to postoperative mediastinitis; obesity (p < 0.001), year of the operation (p < 0.001), low LVEF (p = 0.007) and reoperation for bleeding (p = 0.01) were correlated to superficial infection of the wound and obesity turned out to be the only risk factor for aseptic dehiscence for the sternum (p = 0.003). The infection of the wound did not increase mortality, but it did increase the mean postoperative length of hospital stay (6 days for patients free of any complications of the wound versus 29.7 days for patients with complications of the wound). CONCLUSIONS: In patients with bilateral mammary grafts, obesity is the main risk factor for complications of the wound and this event greatly increases the length of the patient's hospital stay. Consequently, we suggest that bilateral mammary artery grafts be used carefully in this subset of patients. PMID- 9866801 TI - [The clinical, angiographic and procedural predictors of thrombosis and restenosis in Micro stent II (AVE) coronary stents]. AB - The aim of the study was to assess the incidence and the predictors of thrombosis and restenosis in Micro stent II AVE. In a sample of 197 stents successfully implanted in 181 consecutive patients, the incidence of thrombosis was 4.1%. The multivariate analysis showed the minimum lumen diameter post-stenting to be the only independent predictor of overall thrombosis. In fact, we found that the risk of thrombosis increases as the minimal lumen diameter decreases. Angiographic follow-up was available in 74% of the stents at 6.8 +/- 4.1 months and stent restenosis occurred in 26.2% of cases. Independent predictors of restenosis (multivariate linear discriminant analysis) were: 1) nominal stent diameter (the risk of restenosis decreases as the stent diameter increases); 2) the ratio between the diameter of the balloon carrying the stent measured at the maximum pressure/nominal stent diameter (the risk increases as the ratio decreases); 3) stented vessel (the risk increases in the following order: right coronary < circumflex < left anterior descending); 4) the American Heart Association classification of lesion morphology (the risk increases in the order A < B < C); 5) a lower risk was found in the absence of diabetes mellitus. PMID- 9866802 TI - [The prevalence and characteristics of persistent pulmonary hypertension of the newborn. A multicenter study. The Study Group of the Societa Italiana di Cardiologia Pediatrica (SICP)]. AB - BACKGROUND: Persistent pulmonary hypertension of the newborn (PPHN) is a rare syndrome with a severe prognosis, in which a prompt diagnosis can be life-saving. The aim of our study was to verify its prevalence in a neonatal population, define clinical and echocardiographic criteria for the diagnosis of PPHN and discuss therapeutic choices. METHODS: The following clinical and echocardiographic criteria for the diagnosis of PPHN were defined: 1. cyanosis and hypoxemia non-responsive to O2 therapy; 2. right to left shunt at an atrial or ductal level. All neonates fulfilling these criteria referred to the neonatal units of seven pediatric or general hospitals over a two-year period were enrolled. RESULTS: From January 1995 to December 1996, thirty neonates with PPHN (8%) were observed. Birth was pre-term in 5 out of 30. Ten (33%) had experienced chronic and/or fetal asphyxia (FA). Death occurred in 7 (22%), four of whom with FA. Echocardiography showed tricuspid insufficiency in 18 (60%); mean pulmonary systolic pressure was 67 +/- 16 mmHg (range 41-95). In 23 surviving neonates, normalization of clinical and echocardiographic parameters occurred in 8 +/- 5 days. THERAPY: In most neonates, vasodilators (tolazoline, prostacyclin) and/or nitric oxide were employed. CONCLUSIONS: PPHN is confirmed to be a rare pathological condition; prognosis is severe, particularly in neonates with FA. Echocardiography is a reliable non-invasive method for a prompt diagnosis and follow-up. Subsequent studies are needed to assess therapeutic choices. PMID- 9866804 TI - [Models of preoperative risk stratification in cardiac surgery: the prediction of mortality or of survival?]. AB - BACKGROUND: The need to assess the quality of heart surgery outcomes stimulated the development of pre-surgical risk stratification models in order to predict outcome on the basis of patient characteristics. The aim of the study was to compare the predictive accuracy of hospital mortality according to the following three models: Parsonnet (NBI Score), Higgins (CCF Score) and Roques (French Score), in a setting totally independent from the one in which the models were derived. METHODS: For each of the 516 patients undergoing heart surgery at our institution between January 1992 and December 1993, we calculated the pre surgical risk according to the three models. Then we compared the predicted mortality against the observed mortality by means of the Shannon accuracy index, the ROC curve analysis and the overestimation histogram. RESULTS: Overall observed mortality (1.5%) was similar to the predicted mortality by the NBI Score (1.5 +/- 2.1%, p = ns), the CCF Score (1.7 +/- 2.0%, p = ns) and the French Score (1.9 +/- 2.5%, p = ns). The predictive accuracy of global mortality is very high and equal with the three models, and it is very low in the 8 patients who died (NBI Score = 0.06 +/- 0.06; CCF Score = 0.125 +/- 0.067; French Score = 0.102 +/- 0.07, p = ns). The area under the ROC curve is identical in the 3 models. CONCLUSIONS: The predicted mortality obtained by the three models is not significantly different from the observed mortality and therefore, the global accuracy is similar and very high, while it is very low for patients who will die. The models for pre-surgical risk stratification are useful for comparing the results among different institutions or different surgeons, or for monitoring the results over time in the same institution, but they cannot be used to accurately predict the individual risk of hospital mortality. PMID- 9866803 TI - [The effects of the ablation of atrial flutter in patients with and without a clinical history of paroxysmal atrial fibrillation]. AB - BACKGROUND: Although the safety and effectiveness of radiofrequency (RF) transcatheter ablation in patients with atrial flutter (AFL) is well established, little attention is paid to previous history of associated paroxysmal atrial fibrillation (AF) and the recurrence of AFL after RF ablation. In addition, it is not known whether the elimination of AFL can modify the natural history of AF in patients who experience both of these arrhythmias. Accordingly, the aim of this study was to evaluate the effect of RF ablation of AFL in patients with or without a previous history of AF in terms of the incidence of both arrhythmias in the follow-up. METHODS: RF ablation of the atrial isthmus between tricuspid ring, coronary sinus os and inferior vena cava was performed in 27 patients (23 males, 4 females; mean age 61 +/- 9 years) according to the technique described by Cosio. Based on ECG pattern, twenty patients exhibited common or type 1 AFL (negative F waves in the inferior leads with a sawtooth morphology), while seven patients had both common and uncommon AFL (various surface F wave morphologies, generally positive F waves in the inferior leads). A history of association between AFL and paroxysmal AF was documented in 48% of patients, but AFL was the major arrhythmia. After ablation, the patients were followed up and the clinically documented occurrence of arrhythmias was determined. RESULTS: Based on clinical history before ablation, we compared patients with an association between AFL and AF (Gr AFL + AF; n = 13) vs patients with only AFL (Gr AFL; n = 14). The characteristics of the two groups were similar regarding age, sex, duration of symptom, structural heart disease, left atrial size, P-wave duration, AFL interruption during RF procedure, antiarrhythmic treatment before and after RF procedure, and duration of follow-up. During a follow-up of 12 +/- 6 months, AFL recurred in 10 patients (37%), 4 from Gr AFL + AF, and 6 from Gr AFL (p = NS). Episodes of paroxysmal AF occurred in 6 patients (22%), 5 from Gr AFL + AF and 1 from Gr AFL (p < 0.05). In Gr AFL + AF, the incidence of AF after ablation was significantly lower (1.8 +/- 0.6 vs. 0.7 +/- 1 episodes/year; p < 0.02). Characteristics of patients with or without AFL recurrence in the follow-up were similar. The percentage of patients with the occurrence of AFL or AF, associated or unassociated in the follow-up, was 55%. CONCLUSIONS: A history of paroxysmal AF before RF ablation of AFL is not predictive of long-term success or failure of the procedure when considering the recurrence of AFL alone. Nevertheless, the general results are disappointing because the majority of patients have arrhythmias, AFL or AF, associated or unassociated in the follow-up. A clinical history of AF before ablation is correlated with a higher incidence of AF in the follow-up. In any event, the incidence of AF episodes is lower in the follow-up, indicating a possible beneficial effect of AFL ablation on AF mechanisms. PMID- 9866805 TI - [Lipomatous hypertrophy of the interatrial septum: its assessment with TEE, CT and MRI]. AB - BACKGROUND: Lipomatous atrial septal hypertrophy (LHS) is an uncommon condition characterized by the deposition of excessive adipose tissue in the atrial septum. The fossa ovalis is spared by this infiltration of fat. A variety of atrial arrhythmia has been associated with this lesion, which is more common in obese, elderly female patients. Because of the lack of specific symptoms, LHS is the outcome of casual observation during echo-Doppler or autopsy studies. METHODS: We describe the case of a 76-year-old woman who came under our observation due to stroke and atrial arrhythmia. Transesophageal echocardiography clearly revealed lipomatous atrial septal hypertrophy. RESULTS: The MRI confirmed the diagnosis, visualizing the typical "dumbbell" interatrial septum. Computed tomography made it possible to define the anatomic substance of the lesion as "fatty". CONCLUSIONS: Transesophageal echocardiography is the method of choice in the diagnosis of LHS. Other techniques such as RMI or CT-scan can be useful only in selected patients with large fatty infiltration of the atrial wall or marked hypertrophy of the interatrial septum. PMID- 9866806 TI - [An analysis of the decision process in the pharmacological treatment of a patient with chronic heart failure by means of a therapy management information system: the experience of the Montescano Heart Failure Unit]. AB - A computer-based system integrated into the hospital information system for the pharmacological management of the chronic heart-failure patients admitted to the heart-failure unit of Montescano is presented. The major aims of the system involved monitoring the patients' treatment history and the related therapeutic decision-making. The treatment history of the first 151 consecutive CHF patients admitted after the system implementation is examined. The prescribed drug doses at admission and at discharge are compared, as well as the differences in etiology and NYHA class between patients discharged with or without each drug. Several considerations can be drawn from this analysis. First of all, in heart failure patients the choice of drugs is limited and optimal treatment is a result of individualized dosages. Secondly, time and trials are necessary to obtain optimal treatment. Consequently, continuity of care and in- and out-hospital networks are advisable. Third, a computer-based system offers advantages not only for the pharmacological management itself, but also for improving the quality of care through continuous analysis of the decision-making process. PMID- 9866807 TI - [The stratification and prevention of the arrhythmia risk in nonischemic dilated cardiomyopathy]. AB - The challenge of preventing arrhythmic sudden death is one of the major issues in today's treatment of heart failure. To achieve this ambitious goal, an accurate selection of the candidates for sudden death is needed on the one hand, while on the other hand, the assessment of the real cost/benefit ratio of the implantable cardioverter-defibrillator in selected patients, as compared to ACE inhibitors, beta-blockers and antiarrhythmic drug therapy, should no longer be delayed. As is well known, the incidence of sudden death is higher in ischemic dilated cardiomyopathy than it is in non-ischemic dilated cardiomyopathy. Moreover, tachyarrhythmic sudden death is prevalent in NYHA classes I and II (80%), whereas its incidence is lower (50%) in NYHA classes III and IV, since bradyarrhythmia, electromechanical dissociation and thromboembolic events characterize the other 50% of sudden deaths in patients in the latter NYHA class. The stratification of arrhythmic risk in non-ischemic dilated cardiomyopathy is questionable from any point of view, considering the poor predictive power of invasive and non-invasive indexes. However, some subgroups of high-risk patients should be selected, such as patients waiting for heart transplant or those with a severe disease but without an extreme degree of ventricular dysfunction, in whom the prognosis in terms of pump failure events is better and life expectancy is longer if the risk of arrhythmia is properly assessed and sudden death prevented. Consequently, the ICD implant may be effective in order to pursue the aim of reducing the tachyarrhythmic and bradyarrhythmic mortality in patients with a more severe disease and of minimizing the tachyarrhythmic risk in those with a less severe disease. Further studies will be developed to identify the ideal candidates for ICD implants. PMID- 9866808 TI - Acute coronary syndromes: "unifying" theory versus adrenergic stress. PMID- 9866809 TI - [The ablation of chronic atrial fibrillation during mitral surgery: a realistic prospect]. PMID- 9866810 TI - [A remembrance of Guido Chidichimo]. PMID- 9866811 TI - [Hemodynamics laboratories without cine film]. PMID- 9866812 TI - [Hemodynamics laboratories without cine film]. PMID- 9866813 TI - In utero gene therapy: one step closer to reality. PMID- 9866814 TI - A noninvasive prognostic indicator for bladder cancer. PMID- 9866815 TI - Selecting antibodies that block the HIV-1 coreceptor. PMID- 9866816 TI - Restoring interleukin 2 production in AIDS patients. PMID- 9866817 TI - Leukocyte depletion for leukaemia and multiple sclerosis. PMID- 9866818 TI - A new gene Web site for all. PMID- 9866819 TI - Pathogenesis and immunointervention strategies for psoriasis. IBC's Inflammatory Skin Disorders. Washington, DC, USA. September 17-18, 1998. PMID- 9866820 TI - New molecular targets for anticancer therapy. The First International Conference on New Molecular Targets for Anticancer Therapy. Naples, Italy. June 22-23 1998. PMID- 9866821 TI - A piece in the puzzle: an ion channel candidate gene for schizophrenia. AB - Mutations in ion channels have been found to cause a variety of mendelian genetic diseases, and polyglutamine repeat expansion is a newly recognized pathogenic mechanism that causes several rare, genetic, late-onset neurological syndromes. Polymorphic polyglutamine tracts are present in a recently described human, calcium-activated potassium channel, KCNN3 (also known as hKCa3), and alleles of this gene that contain longer repeats have been associated with schizophrenia. The physiological function of the channel is consistent with an etiological role in this disease; drugs designed to target this channel might therefore provide novel psychotherapeutics. PMID- 9866823 TI - Advances in the understanding of early Huntington's disease using the functional imaging techniques of PET and SPET. AB - The functional imaging techniques of positron emission tomography (PET) and single photon emission tomography (SPET) have been used to study regional brain function in Huntington's disease (HD) in vivo. Reduced striatal glucose metabolism and dopamine receptor binding are evident in all symptomatic HD patients and in approximately 50% of asymptomatic adult mutation carriers. These characteristics correlate with clinical measures of disease severity. Reduced cortical glucose metabolism and dopamine receptor binding, together with reduced striatal and cortical opioid receptor binding, have also been demonstrated in symptomatic patients with HD. Repeat PET measures of striatal function have been used to monitor the progression of this disease objectively. In the future, functional imaging will provide a valuable way of assessing the efficacy of both fetal striatal cell implants and putative neuroprotective agents, such as nerve growth factors. PMID- 9866822 TI - Niemann-Pick C disease: cholesterol handling gone awry. AB - Niemann-Pick C disease (NPC) is a debilitating, recessive disorder in humans that causes unrelenting neurological deterioration and is complicated by the presence of lipid-laden foamy cells in the major organs of the body. NPC fibroblasts cultured with an excess of low density lipoprotein (LDL) abnormally sequester cholesterol in their lysosomes. Biochemical analyses of NPC cells suggest an impairment in the intracellular transport of cholesterol to post-lysosomal destinations occurs in NPC. The recent identification of the NPC gene, NPC1, provides a definitive diagnosis of the disease and a means of studying this key component of intracellular cholesterol transport and homeostasis. PMID- 9866824 TI - The role of HLA-B27 polymorphism and molecular mimicry in spondylarthropathy. AB - Ankylosing spondylitis (AS), reactive arthritis (ReA) and other related spondyloarthropathies (SpAs) are characterized by a strong association with the major histocompatibility complex allele HLA-B27. Experimental evidence from humans and transgenic rodents suggests that HLA-B27 is itself involved in the pathogenesis of SpA. Population and peptide-specificity analysis of HLA-B27 suggest it has a pathogenic function related to antigen presentation. Putative roles for infectious agents have been proposed in ReA and suggested in AS. However, the mechanism by which HLA-B27 and bacteria interact to induce arthritis is not clear. Molecular mimicry between bacterial epitopes that cross-react with self-B27 peptides is the most persuasive explanation for the pathogenesis of SpA. The experimental studies reviewed here have greatly increased our knowledge of the structure, function and disease association of HLA-B27. PMID- 9866825 TI - [Effects of newly isolated opioid peptides on G-protein activation: usefulness of [35S] GTP gamma S binding study and its practical application]. AB - Recent cloning and expression studies have revealed that the opioid mu-, delta-, kappa- and orphan receptors are seven-transmembrane domain receptors whose actions are mediated through activation of guanine nucleotide binding protein (G protein). The activation of G-proteins by the opioid receptor can be measured by assessing agonist stimulation of membrane binding of the non-hydrolyzable analog of guanosine triphosphate (GTP), guanosine-5'-O-(3-[35S] thio) triphosphate ([35S] GTP gamma S). Our recent data suggest that 1) the level of spinal mu-, delta-, kappa- and orphan-receptor agonist-stimulated [35S] GTP gamma S binding closely parallels that of receptor binding densities, 2) the neuroanatomical distribution of opioid agonist-stimulated [35S] GTP gamma S binding relates to receptor binding distribution, 3) newly isolated opioid peptides, endomorphin-1 and -2, can activate G-proteins by specific stimulation of mu-receptors and act as partial agonists with moderate catalytic efficacies, 4) mu-receptor densities could be rate-limiting steps in the G-protein activation by mu-agonists in the spinal cord region. In conclusion, opioid agonist-stimulated [35S] GTP gamma S binding can provide a functional method to localize receptors not only by their ability to bind ligands, but also according to their ability to activate an intracellular signal transducer. PMID- 9866826 TI - Pharmacological profiles of absence seizure-induced increases in CRE- and AP-1 DNA-binding activities in gamma-butyrolactone-treated mice. AB - Absence seizures are characterised by a well-defined disturbance of thalamocortical function, and there is no spread to other systems. In this study, we continue our examination of the mechanisms underlying the increased nuclear cyclic AMP responsive element (CRE)- and activator protein 1 (AP-1) DNA-binding activities in a gamma-butyrolactone (GBL)-induced mouse model of absence seizure. The administration of GBL increased CRE- and AP-1 DNA-binding activities in the cerebral cortex and thalamus, but not in other regions such as the hippocampus, cerebellum or pons + medulla oblongata, at doses which induced absence seizures. Not only the absence-seizure behavior but also the increased CRE- and AP-1 DNA binding activities in the thalamocortical regions were reversibly inhibited by ethosuximide, a typical anti-absence drug, and the GABAB antagonists CGP 35348 and CGP 46381. A gel-supershift assay revealed that the GBL-induced CRE-binding activity was supershifted by an anti-CRE-binding protein (CREB) antibody, and that AP-1 DNA-binding activity was blocked by anti-c-Jun and anti-c-Fos antibodies. These results suggest that increased CRE- and AP-1 DNA-binding activities in the cerebral cortex and thalamus are related to the pathogenesis of generalized absence seizures and that these increases in DNA-binding activity are related to ethosuximide- and GABAB antagonist-sensitive abnormal neuronal activity in the thalamocortical circuit. PMID- 9866828 TI - [A case of allergic cross reaction between tricyclic antidepressants and maprotiline]. AB - Although a 76-year-old woman with a diagnosis of depression began a course of maprotiline, the drug was discontinued after 14 days when she developed a rash. She was started on lofepramine in substitution for maprotiline. Four years later, lofepramine treatment was stopped and to amoxapine treatment introduced due to the aggravation of depressive symptoms. On the second day of this regimen, she developed a pruritic rash. She stopped the medication immediately. Because the rash disappeared gradually, lofepramine treatment was restarted. But her recovery was restrained and new skin lesions developed. Again, she stopped the medication, and the rash cleared within several days. This case suggests the existence of cross-allergenicity between tricyclic antidepressants and maprotiline. PMID- 9866827 TI - [Running fit and generalized tonic-clonic seizure are differently controlled by different subtype receptors in the brainstem]. AB - Rats neonatally treated with 0.02% propylthiouracil (PTU) through mother's milk showed a high incidence of audiogenic seizures after maturation. These audiogenic seizures were differently modified by MK-801 and NBQX; while intraperitoneal MK 801 equally inhibited running fit (RF) and generalized tonic-clonic seizure (GTCS), NBQX administered into cisterna ambiens significantly inhibited RF but not GTCS. The possible involvement of glutamate receptors in the inferior colliculus was further investigated using naive Sprague-Dawley rats injected with NMDA, AMPA or cyclothiazide, known as an inhibitor of desensitization of AMPA action. All drugs tested successfully induced RF followed by GTCS, resembling audiogenic seizures in PTU-treated rats. However, sound stimulation could augment AMPA-induced, but not NMDA-induced GTCS. Systemic administration with MK-801 potently blocked GTCS induced by AMPA/cyclothiazide, but the same drug failed to block RF after intracisternal injection with AMPA/cyclothiazide. Furthermore, intracisternal administration with NBQX significantly inhibited only RF induced by AMPA/cyclothiazide. The present study suggests that: 1) glutamate receptors in the brainstem, possible in the inferior colliculus, play a crucial role in audiogenic seizures, namely the initiation of RF and propagation into GTCS; and 2) the initiation mechanism is regulated by both NMDA and AMPA receptors, whereas propagation is mainly controlled by NMDA receptors. PMID- 9866829 TI - [Structure and function of neural plasticity-related gene products]. AB - We have isolated novel immediate early genes (IEGs) from the hippocampus by differential cloning techniques. These mRNAs are induced by synaptic activity and translated into proteins that may affect neural function. We have analyzed a variety of "effector" immediate early genes. These mRNAs encode: 1) cytoplasmic proteins, such as cyclooxygenase-2, a small G protein, Rheb, and a cytoskeleton associated protein, Arc; 2) membrane-bound proteins, such as the cell adhesion protein Arcadlin, and a neurite-outgrowth protein, Neuritin; and 3) a secreted protein, Narp. We hypothesize that physiological stimulation induces "effector" proteins that might strengthen synaptic connections of activated synapses. In contrast, pathological conditions such as epilepsy or drug addiction may accelerate overproduction of these gene products, which cause abnormal synapse formation. Gene targeting and in vivo gene transfer techniques are required to prove this hypothesis. PMID- 9866830 TI - [Structure and function of a novel ErbB ligand, NTAK]. AB - A novel member of the epidermal growth factor (EGF) family, the neural and thymus derived activator for ErbB kinase (NTAK) has been cloned from the cDNA library of a rat pheochromocytoma cell line, PC12 cells and human neuroblastoma cell line, SK-N-SH cells. Four alternative spliced isoforms from rat cDNA have been detected by the methods of RT-PCR. The rat NTAK alpha 2a isoform exhibits 94% identity in its sequence with the human NTAK alpha isoform. Three characteristic Ig-like, EGF like and hydrophobic domains have been identified in rat and human NTAK molecules. Recombinant NTAK, the soluble 46 kDa form, binds directly to ErbB3 and ErbB4, but not ErbB1 and B2. NTAK, however, transactivates with heterodimer such as ErbB1/B3, B1/B4, B2/B3, B2/B4, and B3/B4. NTAK stimulates the differentiation of MDA-MB-453 cells, derived from blast carcinoma. NTAK competitively inhibits the binding of [125I] NRG-1 to these cells. Thus, NTAK is a new member of the EGF family displaying NRG-1 properties. PMID- 9866831 TI - [Analysis of a novel delta gene during the formation of cortical layers]. AB - The preplate is composed of earliest-generated neurons, including Cajal-Retzius cells and prospected subplate neurons which together play a critical role in the formation of cortical layers. Additionally, Notch and Delta have been identified as key components that mediate the lateral specification and fate of numerous cells during development. Here, we demonstrate that a divergent type of Delta (D113) is expressed transiently and specifically in a set of earliest generated neurons in the neural tube. Moreover, in developing cerebrum, D113 is specifically and transiently expressed in prospected subplate neurons. Thus, our results suggest that D113 mediates the specification of cell fate in the earliest stage of neurogenesis including the neurons that compose preplate, leading to the mechanism of genesis of cortical layers. PMID- 9866832 TI - [Genetic alterations in gallbladder carcinoma: a review]. AB - Earlier evidence suggests that gallbladder carcinoma (GBC) has three carcinogenic pathways; de novo development; adenoma-carcinoma sequence; and hyperplasia carcinoma sequence associated with an anomalous arrangement of the pancreaticobiliary duct (AAPBD). We review gene abnormalities in GBC reported to date. p53 mutation and its protein overexpression are frequently observed in de novo carcinoma and GBC with AAPBD, but never found in carcinoma with adenoma. The incidence of K-ras codon 12 mutation in GBCs with AAPBD is significantly higher than that in the other types. Mutation of K-ras is never detected in carcinoma with adenoma. These findings suggest that diverse genetic pathways may exist in gallbladder carcinogenesis and reflect morphologic variations. PMID- 9866833 TI - [Diagnosis of depth of invasion of gallbladder carcinoma with endosonography]. AB - We reported on the usefulness of endosonography (EUS) in the diagnosis of the depth of invasion of gallbladder carcinoma. We performed EUS on 69 patients with gallbladder carcinoma: the degree of tumor extension in 17 was m (tumor invading the mucosa), in 5 pm (tumor invading the muscular layer), in 21 ss (tumor invading the subserosa), 14 in se (tumor invading the serosa), and 12 si (tumor extending to adjacent organs). The detection rate of tumors with EUS was 91.3% (63 of 69). All 6 (100%) patients with pedunculated lesions (Ip type) were correctly diagnosed with EUS. Eighteen of 21 patients (85.7%) with broad-based elevated tumors (Is type), and 5 of 7 (71.4%) with flat-elevated lesions (IIa type) were also diagnosed correctly. EUS is a useful modality for the diagnosis of gallbladder carcinoma. In particular, EUS diagnosis of the depth of invasion is reliable in Ip-type lesions. It is recommended that patients with Ip-type lesions undergo laparoscopic cholecystectomy, but in patients with Is-or IIa-type lesions conventional cholecystectomy with lymph node resection should be performed. PMID- 9866834 TI - [Diagnosis of extension and treatment of carcinoma of the gallbladder]. AB - Diagnosis of extension by intraoperative ultrasonography (IOUS) and treatment based on the degree of histological extension in carcinoma of the gallbladder are discussed. IOUS is a useful technique for the diagnosis of the depth of wall invasion and direct invasion of the liver. The authors diagnose the depth of wall invasion based on the layer structure, unequal width, and discontinuity of the layer echogram. By this technique, mucosal cancer (m cancer) or cancer extending to the proper muscle layer (mp cancer) can be differentiated from cancer with submucosal invasion (ss cancer), and also ss cancer from cancer exposing the serosa (se) or cancer infiltrating to the serosa (si). However, differentiation between m cancer and mp cancer is not possible by IOUS or by other diagnostic techniques. In terms of histological extension, lymph node metastasis or vascular and nerve invasion is not found in m cancer, but in some cases of mp cancer vascular invasion is present. As a radical operative procedure for early m and mp cancer, full-thickness cholecystectomy or partial resection of the liver bed and dissection of lymph nodes 8, 12, and 13 should be conducted. As lymph node metastasis and vascular and nerve invasion are frequent in ss or more advanced cancer, complete lymph node dissection should be performed. Cholecystectomy, partial resection of the liver bed, bile duct resection, and dissection of lymph nodes 8, 12, and 13 is the preferred radical operative procedure for ss cancer. In the cases with in metastasis to lymph nodes 8 and 13, pancreatoduodenectomy is combined. The basic operative procedure for se and si cancer has not been established, but is should be radical and safe. Considering the poor prognosis and frequency of lethal postoperative complications, at present we should not only expand the resected area but select a reasonable and well-balanced operative procedure depending on the degree of cancer extension. PMID- 9866835 TI - [Standard radical surgery for carcinoma of the gallbladder invading the subserosal layer (ss cancer)]. AB - The postoperative survival rate is dependent on the invasive depth of cancer of the gallbladder, as shown in in our studies and in other Japanese studies detailed in questionnaires. In cases of m, mp cancer, good survival rates are achieved after simple cholecystectomy or wedge resection of the liver including the gallbladder bed with regional lymphadenectomy. However, when the cancer has invaded beyond the subserosal layer, lymph node and adjacent organ involvement is found in a large number of patients, and long-term survival cannot be achieved by wedge resection of the liver with dissection of the cystic nodes and pericholedochal lymph nodes. The poor prognosis of patients with these lesions might be due to the use of inappropriate surgical procedures. When the appropriate procedures based on the extent of subserosal (ss) invasiveness of gallbladder cancer were used, long-term survival was obtained in our patient series. The appropriate radical surgical procedures for ss cancer are as follows: When cancer is diagnosed as invading to the ss layer by intraoperative ultrasonography (IOUS), wedge resection of the liver including the gallbladder bed and D2 dissection of the lymph nodes including the paraaortic lymph nodes (16a2 inter, 16b1 inter pre) should be performed. When liver involvement (Hinf1 Hinf2) is diagnosed by IOUS or cancer is located on the liver bed, resection of segment 4a 5 in Couinaud's classification is performed. When there is involvement of the pericholedochal nodes, nodes around the common hepatic artery, and/or posterior pancreatoduodenal nodes are found at operation, pancreatoduodenectomy should be performed. Wedge resection of the liver or S4a 5 segmental resection of the liver with resection of the bile duct and D2 dissection of the lymph nodes including the paraaortic lymph nodes is considered to be the standard radical operation for many patients with ss cancer of the gallbladder. PMID- 9866836 TI - [Hepatic resection for advanced carcinoma of the gallbladder]. AB - Hepatic resection for advanced carcinoma of the gallbladder must be decided upon based on the modes of cancer spread to the liver. The cystic vein through the liver bed is considered an important route of liver metastasis, because liver metastases of gallbladder carcinoma are found frequently around the liver bed. About 70% of early metastatic foci demonstrated microscopically occur in segments 4a and 5. Resection of segments 4a and 5 is considered to be an adequate range of hepatectomy for patients with subserosal invasion, because early metastatic foci are detected not only in patients with direct invasion of the liver but also in those without direct invasion. For patients with direct liver invasion, various degrees of hepatic resection are needed to comply with the depth of direct invasion. It is necessary to achieve negative surgical margins 2 cm from the tumor. Because cancer cells extend along the Glissonian sheath in patients with hilar invasion, extended right hepatectomy with caudate lobectomy is required in these patients. A future problem is to establish the safety of extended hepatectomy in these patients. PMID- 9866837 TI - [Indications for and operative outcome of hepato-pancreatoduodenectomy in the treatment of carcinoma of the gallbladder]. AB - The indications for hepatopancreatoduodenectomy (HPD) are still controversial. Opinion is divided especially concerning the need to perform pancreatoduodenectomy (PD) for lymph node dissection, with some investigators claiming that the peripancreatic lymph nodes can be dissected adequately without PD. Detailed studies of resected specimens in our department have shown that preservation of the head of the pancreas carries an increased risk of leaving behind cancer cells in small lymph nodes and ducts remaining in the peripancreatic region. We therefore perform HPD in all patients at increased risk for metastasis to peripancreatic lymph nodes, as indicated by subserosal invasion with distinct evidence of lymph node involvement, tumor location at the neck or both fundus and body of the gallbladder, and serosa exposure to tumor irrespective of tumor location and lymph node involvement. When the outcome was compared between HPD and hepatic resection plus dissection of peripancreatic lymph nodes (HR), we found that the operative results of HPD were poor with extended lobectomy plus PD because of high rates of postoperative complications and operative mortality. However, the results have gradually improved with the introduction of percutaneous transehepatic portal embolization and advances in perioperative care. Although overall there is no difference between HPD and HR in long-term outcome, the outcome with HPD is significantly better in node-positive patients and patients without hepatoduodenal ligament involvement, there by demonstrating the value of performing peripancreatic lymph node dissection by PD. However, there is no difference between HPD and HR in patients with hepatoduodenal ligament involvement, and attempts to develop other new and effective means of treatment should continue. PMID- 9866838 TI - [Vascular resection during surgical resection for advanced gallbladder carcinoma]. AB - We evaluated the results of vascular resection during surgical resection for advanced gallbladder carcinoma. Twelve patients underwent vascular resection (portal vein in 11, hepatic artery in 2, inferior vena cava in 2) in 58 resected patients with advanced gallbladder carcinoma (stage III and IV). The surgical rate was higher in the nonvascular resection group (61%) than in the vascular resection group (25%) (p < 0.05). Surgical morbidity and mortality rates were not significantly different between the two groups. The survival rate was remarkably higher in the curative resection group (n = 29) (55.6% at 1 year, 30.3% at 3 years, 20.8% at 5 years) than in the noncurative resection group (n = 29) (26.3% at 1 year, 0% at 2 years) (p < 0.05). Survival rates of the nonvascular resection group (n = 46) were 45.3% at 1 year, 23.4% at 3 years, and 16.1% at 5 years. However, no patient in the vascular resection group (n = 12) survived longer than 2 years. In conclusion, vascular resection during surgical resection for advanced gallbladder carcinoma does not result in a more favorable prognosis, despite similar surgical risk as in nonvascular resection procedures. PMID- 9866839 TI - [Value of paraaortic lymphadenectomy for gallbladder carcinoma]. AB - Seven reports of paraaortic lymphadenectomy for advanced carcinoma of the gallbladder were reviewed and positive paraaortic nodes were found in 20-40% of the resected patients and 10-15% of those with subserosal cancer invasion. The rate of patients with positive paraaortic nodes/all patients with positive nodes was 30-50%. Paraaortic lymphadenectomy did not improve the surgical outcome, and most of the patients with positive paraaortic nodes died within 1 year even after aggressive surgery with extensive lymph node dissection. Therefore it is important to clarify the value of paraaortic lymph node dissection for patients with possibly positive paraaortic node metastasis and those with histologically positive nodes excluding the paraaortic area. Although pancreatoduodenectomy for prophylactic lymphadenectomy around the head of the pancreas has been carried out in some institutions, the procedure does not seem to be effective because the main lymphatic route from the gallbladder has a direct connection with the paraaortic nodes via the pericholedochal, periportal, and/or the posterior nodes along the common hepatic artery. The present authors recommend a D2 plus paraaortic lymph node dissection (ext D2) as a standard surgical strategy for carcinoma of the gallbladder. PMID- 9866840 TI - [Laparoscopic splenectomy for a massive splenomegaly using a transcatheter technique]. AB - Laparoscopic splenectomy can be performed more safely today, and therefore it is becoming the first-choice technique for splenectomy when the spleen is of normal size. However, for massive splenomegaly there have been few reports of the use of this technique and its safety has not been confirmed. We performed laparoscopic splenectomy for massive splenomegaly with transarterial embolization of the splenic artery before surgery. A 37-year-old man underwent splenectomy due to the lack of effect of an approximately 4-month course of chemotherapy for chronic myeloid leukemia whose spleen was over 20 cm in length. Before surgery, splenic artery embolization was performed to prevent intraoperative bleeding and to debulk the spleen. Under general anesthesia the patient was positioned in the lateral decubitus position lying on the right side. There was no bleeding from the capsule of the spleen throughout the procedure and no intraoperative complications occurred. Blood loss was 100 ml, and the weight of the resected spleen was 1,100 g. The postoperative course was uneventful. We conclude that laparoscopic splenectomy is safe and feasible in cases of splenomegaly, when combined with preoperative embolization of the splenic artery. PMID- 9866841 TI - [Physiological, pharmacological and molecular aspects of mammalian biological clocks]. AB - Circadian rhythm is an endogenous rhythm that persists in constant conditions with a period of nearly but not identical to 24 hr. Under natural conditions, the circadian clock is precisely entrained to the daily (24 hr) cycle, because environmental stimulus (especially light) induces a phase shift of the clock. In mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus has been shown to be the primary pacemaker that drives daily rhythms of behavioral and physiological activity. Photic information is conveyed from the retina to the SCN directly by the retinohypothalamic tract (RHT) and indirectly by the geniculo hypothalamic tract (GHT). The transmitter of the RHT is glutamate, while the GHT is GABA and neuropeptide Y. Serotonergic innervation from the median raphe and melatonin from the pineal body are likely to provide non-photic information to the SCN. Single gene mutations that dramatically alter circadian phenotype were found in the hamster (tau) and mouse (clock). Moreover, the homologous genes of the Drosophila clock gene, per, were found in mammals and the homologue of the mammalian clock was found in Drosophila. These data suggest that the some constitutes of the biological clock may be conserved between Drosophila and mammals, and a transcription-translation feedback loop involving some clock gene products may be a oscillator itself. PMID- 9866842 TI - [In vivo visualization of renal microcirculation using hydronephrotic rat kidney]. AB - A number of neurohumoral factors participate in the regulation of renal hemodynamics. Several methods have been developed to study directly the regulation of renal microcirculation. Here, we introduce an in vivo method to visualize renal microcirculation by using hydronephrotic rat kidney, a unique method originally developed by Steinhausen et al. More than 10 weeks after unilateral ureteral ligation in rats, the renal parenchyme becomes thinner and suitable for transillumination. After anesthesia, the hydronephrotic kidney was split at the greater curvature with a thermal cautery and then fixed in a water chamber containing Kreb's solution. The renal tissue was transilluminated and microscopically visualized using water immersion objectives. Renal microvessels including arcuate and interlobular arteries, afferent and efferent arterioles and glomerular capillaries could be easily observed on a display monitor at a final magnification of 2,700 times. Topical application of angiotensin II elicited constriction of the interlobular artery and afferent and efferent arterioles dose dependently. Thus, this preparation is a unique model allowing visualization of the whole renal vascular tree in vivo. PMID- 9866843 TI - [Comparative studies on antiparkinsonian agents, talipexole and bromocriptine, evaluated by contralateral rotational behavior in unilaterally nigral-lesioned rats]. AB - The stimulating effect of antiparkinsonian drugs, talipexole and bromocriptine, on the striatal postsynaptic dopamine receptors were studied by measuring contralateral rotational behavior in rats. The nigro-striatal dopamine system of rats was degenerated by unilateral injection of 6-hydroxydopamine (6-OHDA, 8 micrograms/rat) into substantia nigra. By subcutaneous administration, talipexole at 0.16 mg/kg and bromocriptine at 10.24 mg/kg induced significantly increased rotational behavior to the contralateral direction to the lesioned side. The onset of the effect was 30 min for talipexole and 90 min for bromocriptine. By intragastric administration, talipexole at 0.4 mg/kg and bromocriptine at 20.48 mg/kg significantly increased the rotational behavior, and the onset of the effect was 60 min for talipexole and 180 min for bromocriptine. Rotational behavior induced by talipexole was suppressed by a D2 antagonist, sulpiride (40 mg/kg, s.c.), but not by a D1 antagonist, SCH23390 (1 mg/kg, s.c.). In contrast, rotational behavior induced by bromocriptine was suppressed by both sulpiride and SCH23390. These results indicated that when the nigrostriatal dopaminergic functions are disrupted, talipexole stimulates the striatal postsynaptic dopamine receptors at much lower doses than bromocriptine. Also it was indicated that the stimulating effect of talipexole is solely mediated by dopamine D2 receptors, whereas the effect of bromocriptine is mediated by both D1 and D2 receptors. PMID- 9866844 TI - [Effects of amrinone, an inhibitor of c-AMP-specific phosphodiesterases, on neointimal hyperplasia after balloon injury in rats]. AB - Amrinone(5-amino(3,4'-bibyridin)6-(1H)-one), an inhibitor of c-AMP-specific phosphodiesterases, has been used clinically as an inodilator. We have investigated the effect of amrinone on neointimal hyperplasia in rat carotid artery after balloon injury. When amrinone was administered subcutaneously to rats at a dose of 10 mg/kg/day for 14 days, significant reduction of neointimal thickness was noted. Four days treatment with amrinone, however, failed to reduce the intimal thickness. To investigate the mode of action to reduce intimal thickness, we tried to study the effects of amrinone on platelet aggregation and the proliferation of aortic smooth muscle cells. Amrinone produced a dose dependent inhibition of ADP-induced rat platelet aggregation in vitro as well as ex vivo in rats. The proliferation of human aortic smooth muscle cells in culture stimulated with FBS or PDGF was also inhibited by amrinone. These results indicate that amrinone may have potential to prevent vascular neointimal hyperplasia in clinical balloon injury through inhibition of vascular smooth muscle cell growth and platelet aggregation. PMID- 9866845 TI - [The tonsils and adenoids as a site of infection and the cause of obstruction]. AB - The failure to eradicate group A beta-hemolytic streptococci from the pharynx is partly due to a low compliance, but above all, an alteration of the oropharyngeal microbiological flora: reduction of alpha-haemolytic streptococci which inhibit group A beta-hemolytic streptococci and increase of microorganisms such as Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis. These latter act indirectly destroying the beta-lactamic ring of penicillins. However, this obstacle is overcome by the use of antibiotics which do not contain beta lactamic rings such as macrolides or associating amoxicillin with clavulanic acid or with new cephalosporins which are more resistant to beta lactamases. To restrict the diffusion of resistance to antibiotics, it is essential to limit their use diagnosing streptococcal tonsillopharyngitis more precisely, thanks to an improved use of micro-biological diagnostic tests and by a more extended use of tonsillectomy in recurrent tonsillitis (more than 6-7 in 1-2 years). Adenoiditis is closely related to the post nasal drip syndrome, to recurrent otitis media and to otitis media with effusion. All these situations could, therefore, represent an indication, although not well defined, for adenoidectomy. Nasopharyngeal obstruction due to adeno-tonsillar hypertrophy becomes critical during sleep when the hypotony of the upper airway muscles becomes additional to the anatomical obstruction. At this point the inspiratory effort required and the consequent decrease of intra airway pressure increase the pharyngeal obstruction suctioning the pharyngeal walls toward the median line. The resulting clinical picture is defined as sleep-disordered breathing (SDB) due to adenotonsillar hypertrophy (idiopathic), to be distinguished from SDB due to cranio-facial abnormalities or neuromuscular diseases. SDB includes both the more serious sleep apnea syndrome and the less severe upper airway respiratory resistance syndrome. A combination of symptoms and clinical data detectable both while awake or asleep, make the diagnosis simple. During sleep, both apnea and paradoxical inspiratory movements are highly specific while snoring is highly sensitive. To evaluate nasopharyngeal obstruction radiography and optic fibre endoscopy are both equally reliable. The gold standard test for non idiopathic SDB is the polysomnography, whereas for SDB, due to adenotonsillar hypertrophy, one is limited today to the recording during sleep of O2 saturation or of end tidal CO2. These investigations are, however, generally used up to 2 years of age, when the decision to carry out an adenoidectomy and especially a tonsillectomy is more difficult because of the greater risks which surgery involves at this age. The pharmacological therapy has a purely palliative function and is based on antibiotics, local vasoconstrictors, steroids and theophylline which acts more as an antiflogistic than as a breath stimulant. O2 therapy and nasal continuous positive airway pressure (CPAP) give better results, but are more difficult to carry out, in particular on a long term basis. Adenoidectomy especially if associated with tonsillectomy, leads to the resolution of the symptoms, but not always to a normalization of functional alterations (hypoxia and hypercapnia). For this reason, it is necessary to act on other factors which cause oedema of the nasopharyngeal mucosa contributing to the obstruction. In this area, the prevention of viral infections can be achieved by vaccination against influenza and by preventing the child from attending crowded day care centers. With regard to allergic inflammation, skin prick tests could be a first step in view of allergens avoidance measures. With regard to indoor air pollution, passive smoke must be stopped and the child kept out of the kitchen. PMID- 9866846 TI - [Adolescents and suicide. Some comments on the role of the pediatrician in researching the motivations for suicidal behaviors]. AB - The modern pediatric is interested also in the adolescent. Sometimes disorders of the behaviour lived of a teenager in a family on risk can lead to attempts of suicide. The authors will analyze at the basis of their experience like pediatrics this problematic. PMID- 9866847 TI - [The lexical aspects in children with expressive language delay]. AB - This study examines the lexical characteristics of language development of 17 children between 24 and 41 months of age with a history of slow expressive language acquisition (experimental group). The purpose of the investigation is to compare these subjects to 49 children (ranging in age from 13-24 months) with a normal language history matched for vocabulary size to determine, whether the expressive vocabulary of children with language delay is same to normal group. The data were collected using the parental report instrument "Il Primo Vocabolario del Bambino"(Caselli and casadio 1995) the Italian version of MacArthur Communicative Development Inventories (Fenson et al. 1993). For analyses of word production, children were divided into four groups based on their total vocabulary size: 1) 11 to 20 words; 2) 21 to 50 words; 3) 51 to 100 words; 4) 101 to 200 words. Analysis revealed a significant difference in verbs and grammatical function words when the level of vocabulary size of experimental group was between 21 and 50 words. In the other vocabulary size there were not significant differences. In summary the results of this study show that the vocabulary composition of children with slow expressive language acquisition is same that of children 14-16 months younger. PMID- 9866848 TI - [Growth deficiency and renal tubular acidosis. The possible etiopathogenic mechanisms]. AB - The authors have studied two cases of renal tubular acidosis in 2 children underlining that a possible defective mechanism of growth is based on alteration of the receptors of somatomedine C to level of cartilage. The deficit of receptors could be the consequence of the alterations of the acidosis on cartilage. PMID- 9866849 TI - [Calcium phosphate metabolism in thalassemia]. AB - The AA. performed a screening test on 113 patients affected by beta thalassemia major ranging between 2 and 40 years of age, randomized among those who come to the Microcitemic Center of our Institute, and on a control group. In all of them serum levels of calcium, phosphate, parathyroid hormone (PTH), calcitonin and 25 OH vitamin D were measured. Average serum levels of PTH were significantly (P < 0.001) lower in our patients than in control group and 12.4% of the former were clearly under normal range, especially in the group over 16 years of age. Also serum levels of 25-OH vitamin D were lower in thalassemic patients than in controls, because of the presence of 32 patients with average values under normal limit. Our results are in agreement with current literature and underline the increasing incidence of endocrine complications in thalassemic patients who undergo high transfusion regimens, because of to the increase of hemosiderosis due to the low compliance to iron chelation therapy. Controversial is the pathogenesis of the absence of hypocalcemia in many patients with hypoparathyroidism and the cause of the deficit of vitamin D. PMID- 9866850 TI - [Renal polycystosis in pediatrics]. AB - Inherited polycystic kidney disease (PKD) in children is a disorder characterized by diffuse cystic involvement of both kidneys, without evidence of dysplasia. Both forms, autosomal recessive (ARPKD) and autosomal dominant (ADPKD) have considerable overlap in clinical presentation and radiographic features in the pediatric population. At present, a prenatal diagnosis is possible with ultrasound examination. A brief review of the literature is here reported. The knowledge of pathophysiological and clinical data is requested, since PKD represents a major cause of renal failure in pediatrics. PMID- 9866851 TI - [Collagen diseases and the involvement of the mouth in childhood]. AB - In the review, the authors describe the lesions of the oral cavity, which are present in lupus erythematosus, scleroderma, dermatomyositis. They underline the importance of their knowledge for the dentist and for the pediatrician, because the last recognition can influence prognosis negatively. PMID- 9866852 TI - [Varicocelectomy by an inguinal approach. Our experience]. AB - Varicocele is a dilatation of the spermatic plexus due to a pathologic venous reflux in the testes. It affects about 15-18% of adolescents. The modern diagnostic tools allow an early identification and a thorough staging. Surgical treatment in this phase seems to be useful for the preservation of gonad integrity. Recently, the interest in varicocele in pediatric surgery has increased because of the close relationship of this disease to male infertility. We performed a diagnostic-therapeutic protocol with a follow-up to evaluate the trophism and functional state of testes and to identify early recurrence of venous reflux. In our Institute, the surgical treatment usually performed is resection of spermatic plexus through an inguinal approach. This technique is simple, yields good outcome and has no morbidity. PMID- 9866854 TI - [Neonatal chylous ascites: a case report and review of the literature]. AB - The case of a newborn with asymptomatic chylous ascites operated on because of voluminous inguino-scrotal hernia is reported and a review of the recent literature about aetiology, diagnosis and treatment of neonatal chylous ascites is made. PMID- 9866853 TI - [Cheilognathopalatoschisis: its embryology, epidemiology, etiology, clinical picture and therapy. The experience of a division of pediatrics]. AB - The authors report their experience about cleft lip and palate. PMID- 9866855 TI - [Transient pseudohypoaldosteronism secondary to congenital malformation pathology of the urinary tract (valves of the posterior urethra): a report of a case with elements of a physiopathological nature]. AB - A case of secondary transient pseudohypoaldosteronism (PHA) in a three-month-old infant who had arrived in emergency conditions is reported. Clinically the patient presented dehydration associated with salt loss, metabolic acidosis, severe hyperkalemia, markedly elevated levels both of plasma renin activity and aldosterone concentration. Biochemical and instrumental tests and clinical evolution have given reason to consider it as a case of transient PHA, secondary to an obstructive uropathy (PUV) with urinary infection. Secondary forms of PHA have frequently been documented in young infants with urinary tract infection associated with both obstructive and non obstructive uropathy. After medical or surgical therapy the abnormalities quickly disappear. This reported case suggests that the pediatrician-neonatologist dealing with a young infant with obstructive uropathy should not only consider the surgical treatment but also the medical one, in the light of the associated physiopathological aspects. PMID- 9866856 TI - [The multiple pterygium syndrome or Escobar syndrome: a case report]. AB - The authors report on a new case of multiple pterygium syndrome (MPS) or Escobar's syndrome. Generally the seriousness of MPS is due to the severity of the flexion deformities of the joints and to the serious and evolutionary deformities in the spine. Our case is different from classical description of MPS for absence of spine deformities and for presence of bony lesions below the pterygia, for a bizarre aspect of the ears and for foot deformities. PMID- 9866857 TI - [Microcirculation, tissue perfusion and oxygen transfer in the newborn, nursing and healthy infant and in intensive therapy]. PMID- 9866858 TI - Protein engineering the surface of enzymes. AB - The protein surface is the interface through which a protein senses the external world. Its composition of charged, polar and hydrophobic residues is crucial for the stability and activity of the protein. The charge state of seven of the twenty naturally occurring amino acids is pH dependent. A total of 95% of all titratable residues are located on the surface of soluble proteins. In evolutionary related families of proteins such residues are particularly prone to substitutions, insertions and deletions. We present here an analysis of the residue composition of 4038 proteins, selected from 125 protein families with < 25% identity between core members of each family. Whereas only 16.8% of the residues were truly buried, 40.7% were > 30% exposed on the surface and the remainder were < 30% exposed. The individual residue types show distinct differences. The data presented provides an important new approach to protein engineering of protein surfaces. Guidelines for the optimization of solvent exposure for a given residue are given. The cutinase family of enzymes has been investigated. The stability of native cutinase has been studied as a function of pH, and has been compared with the cutinase activity towards tributyrin. Whereas the onset of enzymatic activity is linked with the deprotonation of the active site HIS188, destabilization of the 3D structure as determined by differential scanning calorimetry is coupled with the loss of activity at very basic pH values. A modeling investigation of the pH dependence of the electrostatic potentials reveals that the activity range is accompanied by the development of a highly significant negative potential in the active site cleft. The 3D structures of three mutants of the Fusarium solani pisi cutinase have been solved to high resolution using X-ray diffraction analysis. Preliminary X-ray data are presented. PMID- 9866859 TI - Biodegradable products by lipase biocatalysis. AB - The interest in the applications of biocatalysis in organic syntheses has rapidly increased. In this context, lipases have recently become one of the most studied groups of enzymes. We have demonstrated that lipases can be used as biocatalyst in the production of useful biodegradable compounds. A number of examples are given. 1-Butyl oleate was produced by direct esterification of butanol and oleic acid to decrease the viscosity of biodiesel in winter use. Enzymic alcoholysis of vegetable oils without additional organic solvent has been little investigated. We have shown that a mixture of 2-ethyl-1-hexyl esters can be obtained in a good yield by enzymic transesterification from rapeseed oil fatty acids for use as a solvent. Trimethylolpropane esters were also similarly synthesized as lubricants. Finally, the discovery that lipases can also catalyze ester syntheses and transesterification reactions in organic solvent systems has opened up the possibility of enzyme catalyzed production of biodegradable polyesters. In direct polyesterification of 1,4-butanediol and sebacic acid, polyesters with a mass average molar mass of the order of 56,000 g mol-1 or higher, and a maximum molar mass of about 130,000 g mol-1 were also obtained by using lipase as biocatalyst. Finally, we have demonstrated that also aromatic polyesters can be synthesized by lipase biocatalysis, a higher than 50,000 g mol-1 mass average molar mass of poly(1,6-hexanediyl isophthalate) as an example. PMID- 9866860 TI - Studies on kinetic parameters and stability of aminoacylase in non-conventional media. AB - Catalytic properties and conformational stability of aminoacylase (N-acylamino acid amidohydrolase, EC 3.5.1.14) were studied in water-N,N-dimethylformamide (DMF) and water-dioxane solvent mixtures. Beside the prompt inhibition the solvents caused further inactivation during incubations. In the presence of 5% DMF content the inactivation proceeds with a well-measurable rate (t1/2 39 min), while in the case of 20% DMF the enzyme practically lost its starting activity during 50 min incubation (t1/2 13 min). The K(m) value of the enzyme increased about three times with increasing DMF concentrations up to about 2.6 M DMF, while the Vmax value decreased practically to zero in the same concentration range. PMID- 9866861 TI - Carboxypeptidase A-catalyzed dipeptide synthesis in organic media. AB - The carboxypeptidase A-catalyzed syntheses of dipeptides from L-amino acids (Phe, Tyr, Trp, Leu and Ile) were studied in various water-miscible (acetone, acetonitrile, ethanol, methanol and 1,4-dioxane) organic solvents. The highest yield (43%) was achieved in acetonitrile with L-Phe as substrate, after a 24-h incubation. The optimal conditions of Phe-Phe synthesis in acetonitrile were determined. For maximal conversion 1.2 mM L-Phe, 1.4 mg ml-1 enzyme and about 10% water are needed in buffered aqueous acetonitrile (pH 5.5) at 30 degrees C. PMID- 9866862 TI - Perspectives in agrobiotechnology. AB - This review surveys the most important and promising contributions of agricultural biotechnology to the development of sustainable, environment friendly agriculture. It deals with the recent achievements of genetic technology for the development of new transgenic microbial, plant and animal products. It also deals with the newest developments and perspectives of microbial intervention in agricultural practices, such as biofertilizers, biocontrol agents, and various microbiological products used in modern agriculture. The review surveys the outlook for a waste-free, environment-friendly sustainable agricultural practice, including waste management, recycling and bioremediation technologies. The review lists the most important marketable agrobiotechnological products, and their present and projected sales volume. PMID- 9866863 TI - Mycotechnology: the role of fungi in biotechnology. AB - Fungi have been important in both ancient and modern biotechnological processes. Processes and products that utilize fungi include baking, brewing, and the production of antibiotics, alcohols, enzymes, organic acids, and numerous pharmaceuticals. The advent of recombinant DNA technology and large scale genomics analysis has placed yeasts and filamentous fungi in the forefront of contemporary commercial applications. The term 'mycotechnology' is introduced here to describe the enormous impact of fungi on biotechnology. PMID- 9866864 TI - Cation-exchange displacement chromatography for the purification of recombinant protein therapeutics from variants. AB - Removal of low level impurities that are closely related to the bioproduct is a commonly encountered challenge in the purification of biopharmaceuticals. These separations are typically carried out by using shallow linear salt gradients at relatively low column loadings, significantly limiting the throughput of the purification process. In this manuscript we examine the utility of displacement chromatography for the purification of recombinant human brain-derived neurotrophic factor, rHuBDNF. The utility of displacement chromatography is compared to gradient elution for the removal of variants of the rHuBDNF. The results demonstrate that displacement chromatography is capable of achieving high yields and purity at high column loadings. Displacements developed on 20 microns and 50 microns cation-exchange resins are shown to provide 8-fold and 4.5-fold increases in production rates, respectively, when compared to an existing linear gradient elution operation. These results demonstrate the efficacy of displacement chromatography for the purification of therapeutic proteins from complex feed streams. PMID- 9866866 TI - Functional expression of Rhizopus oryzae lipase in Pichia pastoris: high-level production and some properties. AB - The mature lipase of the fungus Rhizopus oryzae (ROL) was functionally expressed and secreted in the methylotrophic yeast Pichia pastoris. In a batch cultivation, where methanol feeding was linked to the dissolved oxygen content in the cultivation solution, a lipase activity of 500,000 units per liter (60 mg active lipase per liter) of culture was achieved after initial glycerol feeding of the culture. Recombinant ROL lipase was purified to homogeneity by a simple two-step purification procedure and had a specific activity of 8571 U mg-1 (triolein, 30 degrees C, pH 8.1) which is comparable with the purified native enzyme. The properties of the recombinant lipase were similar to those reported both for the native lipase and for the enzyme expressed in Escherichia coli and refolded from inactive inclusion bodies. PMID- 9866865 TI - The conformation of purified Toxoplasma gondii SAG1 antigen, secreted from engineered Pichia pastoris, is adequate for serorecognition and cell proliferation. AB - A truncated form of SAG1, the immunodominant surface antigen of Toxoplasma gondii, has been produced in the methylotrophic yeast, Pichia pastoris. By construction, the recombinant protein lacks C-terminal residues 308-336 which, in native SAG1, encompass the glycosylphosphatidylinositol anchorage site. Secretion of anchor-less SAG1 proceeded via the yeast prepro alpha-mating factor signal peptide and yielded two immunoreactive protein species having apparent molecular masses of 31.5 and 34.5 kDa, respectively, and differing only by N-glycosylation of the single Asn-X-Ser site present in the molecule. Purification of the anchor less SAG1 was achieved by a combination of ion-exchange and size-exclusion chromatographies. N-terminal amino acid sequencing of the products indicated the presence of additional residues glutamic acid--alanine at the N-terminal end of the products. Despite incomplete processing and unnatural glycosylation, anchor less SAG1 proteins apparently adopted a suitable conformation recognized by monoclonal and human serum-derived antibodies, specific for the native SAG1. In addition, the recombinant anchor-less SAG1 proved competent for inducing proliferation, in vitro, of mononuclear cells from seropositive individuals. Finally, properly adjuvanted anchor-less SAG1 was able to induce protection of mice against a lethal challenge with T. gondii tachyzoites. PMID- 9866867 TI - Quantitative assay system for specific enzyme activity using antibody: the case of protease, subtilisin BPN'. AB - Assay on a high-quality microtiter plate was found to allow for quantitative analysis of bacterial serine protease, subtilisin BPN', and its mutant enzymes which had been genetically engineered to be adapted to low-temperatures (Taguchi et al., 1998. Appl. Environ. Microbiol. 64, 492-495), by using polyclonal antibody against subtilisin BPN'. The use of polyclonal antibody was crucial in normalizing the number of various different enzyme molecules in culture supernatant samples of recombinant strains of Bacillus subtilis, giving rise to the performance of specific activity assay of the enzymes. Relative activity of each mutant subtilisin BPN' to wild-type enzyme was estimated by monitoring the increased value of absorbance caused by enzymatic hydrolysis of a chromogenic substrate. The relative activity in each enzyme estimated by this method showed good coincidence with that estimated by kinetic parameters, kcat/K(m) of the purified enzymes. We termed the system as 'ABEA' (antibody-bound enzyme assay). The efficient ABEA system developed here would be useful for the determination of specific activity of other enzymes of interest and provide us versatile applications in the field of evolutionary engineering. PMID- 9866869 TI - Expression of the second epidermal growth factor-like domain of human factor VII in Escherichia coli. AB - The second epidermal growth factor (EGF)-like domain of human coagulation factor VII is a potent inhibitor of the FVIIa/tissue factor complex, the predominant initiator of coagulation in vivo. This domain has now for the first time been cloned and expressed in Escherichia coli as an affinity fusion protein. The fusion protein was secreted into the periplasm of E. coli and purified by affinity chromatography. The purified protein consisted of a fusion protein with the expected molecular weight, and in addition, a significant fraction of oligomers cross-linked by intermolecular disulfide bonds. Despite the presence of oligomers, the purified protein was a potent inhibitor of the extrinsic blood coagulation pathway with an IC50 value of about 20 microM. The biological activity was retained after liberation of the EGF domain by proteolytic cleavage. PMID- 9866868 TI - Preparation, characterization and application of Aspergillus sp. xylanase immobilized on Eudragit S-100. AB - Aspergillus sp. 5 (strain 5) and Aspergillus sp. 44 (strain 44) produced xylanase (34.3 and 32.7 IU ml-1, respectively) with very low levels of cellulases when grown on 1% wheat bran medium. Xylanase was non-covalently immobilized on Eudragit S-100 for saccharification. The system retained 70 and 80% of strain 5 and strain 44 xylanase activity, respectively. On immobilization, optimum temperature of activity broadened between 50 and 60 degrees C as compared to 50 degrees C in the case of the free enzymes. No significant shift in the pH optima was observed on immobilization. However, immobilization increased enzyme stability mainly by decreasing the temperature sensitivity to the inactivation reaction. The K(m) values increased from 5.6 to 8.3 mg ml-1 for strain 5 xylanase and 7.0 to 9.0 mg ml-1 for strain 44 xylanase. Enzymatic saccharification of xylan and wheat bran was improved on xylanase immobilization. Immobilized xylanase from both the strains produced three times more sugar as compared to free xylanase. In repeated batch saccharification studies immobilized xylanase was recycled three times without loss of enzyme activity. PMID- 9866870 TI - Heterologous phi X174 gene E-expression in Ralstonia eutropha: E-mediated lysis is not restricted to gamma-subclass of proteobacteria. AB - E-lysis of Ralstonia eutropha H16, which belongs to the beta-subclass, was undertaken to verify whether transmembrane tunnel formation is possible in bacteria which do not belong to the enterobacteriaceae. For this purpose, a new gene E expression plasmid, pKG12, with two origins of replication, oriV and oriT, from plasmid pRP4, chloramphenicol and kanamycin resistance genes and a casette composed of lambda cI857 and lambda pR gene E was constructed. Temperature upshift of R. eutropha H16 (pKG12) from 28 to 45 degrees C during exponential growth resulted in lysis of the strain with features characteristic of E-mediated lysis of Escherichia coli. The cytoplasmic contents released can easily be separated from the still intact envelope fraction by centrifugation or filtration. As R. eutropha H16 represents an important industrial organism, E mediated lysis could facilitate procedures for the recovery of intracellular mediators or products like polyhydroxyalkanoates. PMID- 9866871 TI - Towards an earlier diagnosis of Alzheimer's disease. PMID- 9866872 TI - Evolutionary link between prokaryotic and eukaryotic K+ channels. AB - Considering the importance of K+ channels in controlling the crucial K+ gradient across the plasma membranes of all living cells, it comes as no surprise that, besides being present in every eukaryotic cell, these integral membrane proteins have recently also been identified in prokaryotes. Today, approximately a dozen successfully completed and many more ongoing sequencing projects permit a search for genes related to K+ channels in the genomes of both eubacteria and archaea. The coding regions of homologues show a remarkable variety in primary structure. They predict membrane proteins with one, two, three and six hydrophobic segments surrounding a putative K(+)-selective pore (H5) and the presence or absence of a cytosolic putative NAD(+)-binding domain (PNBD) that probably senses the reducing power of the cell. The analysis of kinships on the basis of phylogenetic algorithms identifies sequences closely related to eukaryotic voltage-dependent Kv channels, but also defines members of a primordial class of prokaryotic K+ channel (containing the 2TMS/PNBD motif). Considering the unique mechanisms that may account for the assembly of modern proteins from different ancestral genes, and with more primary sequence data soon to appear, a scheme for the evolutionary origin of K+ channels comes within reach. PMID- 9866873 TI - Molecular cloning and developmental expression patterns of the MyoD and MEF2 families of muscle transcription factors in the carp. AB - cDNA clones encoding the myogenic regulatory factors (MRFs) myogenin, MyoD and myf-5 were isolated by reverse-transcription polymerase chain reaction from larvae and embryos of the common carp (Cyprinus carpio L.). Myocyte-specific enhancer factor 2 (MEF2) cDNAs were identified from a cDNA library from adult carp. Northern blot analysis showed that MyoD, myf-5 and MEF2C transcripts were present in three-somite embryos, whereas myogenin and MEF2A transcripts were not detected until the 15-somite stage. Intense signals of myogenin and MyoD transcripts were observed even in 1-month-old juveniles. Levels of MyoD, myogenin and MEF2A transcripts declined between 1 and 7 months after hatching, and myf-5 gave only a weak signal in the oldest fish. In contrast, levels of MEF2C transcripts were considerably higher in 7-month-old juveniles than in 1-month-old larvae. mRNAs encoding carp myosin heavy chain and alpha-actin were first detected at approximately the time of the first heartbeat, and levels were maximal in juveniles 1 month post-hatching. The relatively high levels of MRF mRNA in juvenile fish probably reflect the recruitment of new muscle fibres from the satellite cell population. It was concluded that the relative importance of the different members of the MyoD and MEF2 families of transcription factors for muscle differentiation changes during ontogeny in the carp. PMID- 9866874 TI - Two types of mRNA encoding myosin regulatory light chain in carp fast skeletal muscle differ in their 3' non-coding regions and expression patterns following temperature acclimation. AB - cDNA clones encoding the myosin regulatory light chain (RLC) were isolated from a cDNA library prepared from fast skeletal muscle of the carp Cyprinus carpio L. Two types of cDNA clone encoding carp RLC were found with identical deduced amino acid sequences. The two mRNAs differed in the number of polyadenylation signals prior to the poly(A) tail in the 3' non-coding region. The two mRNA species, with approximate sizes of 1.4 and 0.8 kilobases, were also observed in northern blot analysis. Carp were acclimated for a minimum of 5 weeks to either 10 degrees C or 30 degrees C (14h:10h light:dark photoperiod). The total levels of mRNA transcripts coding for the RLC and myosin heavy chain were, respectively, 3.3 and 3.9 times higher in cold-than in warm-acclimated fish. Differences in the levels of RLC in mRNA transcripts were largely due to the concentration of the 1.4 kilobase mRNA species. PMID- 9866875 TI - Comparative kinematics and hydrodynamics of odontocete cetaceans: morphological and ecological correlates with swimming performance. AB - Propulsive morphology and swimming performance were compared for the odontocete cetaceans Delphinapterus leucas, Orcinus orca, Pseudorca crassidens and Tursiops truncatus. Morphological differences were apparent among the whales. The general body contour and low-aspect-ratio caudal flukes of D. leucas indicated that this species was a low-performance swimmer compared with the other species. Propulsive motions were video-taped as animals swam steadily in large pools. Video tapes were analyzed digitally using a computerized motion-analysis system. Animals swam at relative velocities ranging from 0.4 to 2.4 body lengths s-1. The stroke amplitude of the flukes decreased linearly with velocity for D. leucas, but amplitude remained constant for the other species. Tail-beat frequencies were directly related to relative swimming velocity, whereas the pitch angle of the flukes was inversely related to relative swimming velocity. Unsteady lifting-wing theory was used with regression equations based on kinematics to calculate thrust power output, drag coefficients and propulsive efficiency. Compared with other species, O. orca generated the largest thrust power (36.3 kW) and had the lowest drag coefficient (0.0026), whereas T. truncatus displayed the largest mass specific thrust power (23.7 W kg-1) and P. crassidens had the highest efficiency (0.9). D. leucas did not swim as rapidly as the other species and had a comparatively higher minimum drag coefficient (0.01), lower mass-specific thrust power (5.2 W kg-1) and lower maximum efficiency (0.84). Minimum drag coefficients were associated with high swimming speeds, and maximum efficiencies corresponded with velocities in the range of typical cruising speeds. The results indicate that the kinematics of the propulsive flukes and hydrodynamics are associated with the swimming behaviors and morphological designs exhibited by the whales in this study, although additional factors will influence morphology. PMID- 9866876 TI - Mechanics of lung ventilation in a larval salamander Ambystoma tigrinum. AB - The larval stage of the tiger salamander Ambystoma tigrinum is entirely aquatic, but the larvae rely on their lungs for a large proportion of their oxygen uptake. X-ray video and pressure measurements from the buccal and body cavities demonstrate that the larvae inspire using a two-stroke buccal pump and exhale actively by contracting the hypaxial musculature to increase body pressure. Larvae begin a breath by expanding the buccal cavity to draw in air through the mouth, while simultaneously exhaling air from the lungs to mix with the fresh air in the buccal cavity. The mouth then closes, and the buccal cavity compresses to pump a portion of the mixture into the lungs. The remaining air in the buccal cavity is then released as bubbles from the mouth and gill slits. Ventilatory volumes estimated from X-ray video records indicate that approximately 80% of the air pumped into the lungs is fresh air and 20% is previously expired air. Exhalation in larval tiger salamanders is active, powered by contraction of all four layers of lateral hypaxial musculature. Electromyography indicates that the transverse abdominis (TA) muscle is active for the longest duration and shows the highest-amplitude activity, but the external oblique superficialis, the external oblique profundus and the internal oblique also show consistent, low-level activity. The finding that the TA muscle is active during exhalation in larval tiger salamanders contributes to a growing body of evidence that the use of the TA for exhalation is a primitive character for tetrapods. PMID- 9866877 TI - Structure and function of the vomeronasal organ. AB - Many animals use their vomeronasal organs to gain direct and specific contact with chemical cues released by congeners and in biological fluids. These cues provide information about the physiological status of the emitter and facilitate or regulate social interactions such as sexual relationships. The present review gives a short description of the discovery of the vomeronasal organ and the pivotal findings of Jacobson. The distribution of the organ and its anatomy in some vertebrates are described. The mechanisms for stimulus entry and egress are discussed, and the findings that led to the appreciation of the vomeronasal organ in mammals as a main chemosensory organ for pheromones mediating reproductive status and inducing sexual behaviour are reported. The anatomical, biochemical and functional properties of the receptor neurones are described. PMID- 9866878 TI - Determinants of the center of mass trajectory in human walking and running. AB - Walking is often modeled as an inverted pendulum system in which the center of mass vaults over the rigid stance limb. Running is modeled as a simple spring mass system in which the center of mass bounces along on the compliant stance limb. In these models, differences in stance-limb behavior lead to nearly opposite patterns of vertical movements of the center of mass in the two gaits. Our goal was to quantify the importance of stance-limb behavior and other factors in determining the trajectory of the center of mass during walking and running. We collected kinematic and force platform data during human walking and running. Virtual stance-limb compression (i.e. reduction in the distance between the point of foot-ground contact and the center of mass during the first half of the stance phase) was only 26% lower for walking (0.091 m) than for running (0.123 m) at speeds near the gait transition speed. In spite of this relatively small difference, the center of mass moved upwards by 0.031 m during the first half of the stance phase during walking and moved downwards by 0.073 m during the first half of the stance phase during running. The most important reason for this difference was that the stance limb swept through a larger angle during walking (30.4 degrees) than during running (19.2 degrees). We conclude that stance-limb touchdown angle and virtual stance-limb compression both play important roles in determining the trajectory of the center of mass and whether a gait is a walk or a run. PMID- 9866880 TI - The effect of respiratory pattern on water loss in desiccation-resistant Drosophila melanogaster. AB - We measured CO2 and H2O release from individual fruit flies from five populations of Drosophila melanogaster selected for resistance to desiccation (D flies). Our previous work found that these flies survive for an extended period in dry air, have an increase in the peak height and frequency of CO2 release, as measured by the standard error of a linear regression (SER) of CO2 release for the entire survival period, and have reduced water loss rates (VH2O) compared with their control or ancestor populations. In the present study, we examined the following respiratory characteristics: VCO2, VH2O, the SER of CO2 release and the ratio of VCO2 to VH2O in the D flies. Correlations between these characters were calculated in order to determine the effect of respiratory pattern on water loss. We found that, within the D flies, neither periodic release of CO2 nor an increased SER for CO2 release was associated with reduced water loss. In addition, an increased SER was positively correlated with both an increased water loss rate and a decreased survival time. Therefore, although selection for desiccation resistance leads to both an increased SER and a decreased rate of water loss in the D flies, the increased SER does not significantly reduce respiratory water loss. PMID- 9866879 TI - Using laboratory selection for desiccation resistance to examine the relationship between respiratory pattern and water loss in insects. AB - We conducted concurrent measurements of rates of CO2 and H2O release from individual fruit flies Drosophila melanogaster taken from populations subjected to three different selective regimes: (1) populations selected for resistance to desiccation (D flies); (2) populations maintained as their controls (C flies); and (3) the ancestral populations of the D and C populations (O flies). In the D flies, water loss rates were significantly reduced, the standard error of the regression (SER) of the CO2 release pattern measured over the survival period of the flies was increased, and the ratio of CO2 loss to H2O loss (VCO2/VH2O) was increased. Correlations across all 15 populations from the three selection treatments indicate that survival time was negatively correlated with water loss rate, positively correlated with the SER of CO2 release and positively correlated with the VCO2/VH2O ratio. We did not, however, find a significant correlation between the SER of CO2 release and rates of water loss or the VCO2/VH2O ratio. PMID- 9866881 TI - Effects of food nutrient content, insect age and stage in the feeding cycle on the FMRFamide immunoreactivity of diffuse endocrine cells in the locust gut. AB - We have studied the influence of variations in dietary protein and digestible carbohydrate content, of insect age and of time during the feeding cycle on the endocrine cells of the ampullar region of the midgut in the African migratory locust Locusta migratoria L. Morphometric analysis of FMRFamide-like immunoreactivity was used as an indirect measure of the amount of FMRFamide related peptides (FaRPs) stored in the gut endocrine cells. There was a highly significant correlation between FaRP content and the nutritional quality of the food, measured relative to the concentrations and ratio of protein to digestible carbohydrate in a nutritionally optimal diet. The direction of the relationship between FaRP content and diet quality varied with age during the fifth stadium. On day 1, FaRP levels increased with the nutritional quality of the food, while on day 4 the opposite relationship was observed. Release of peptide was triggered by the onset of a meal during ad libitum feeding, with cell FaRP levels returning to premeal values within 15 min of the meal ending. The results also suggested that cell contents were released during food deprivation beyond the normal intermeal interval. Locusts switched for a single meal during ad libitum feeding on day 4 from a low- to a high-carbohydrate food did not respond by reducing endocrine cell FaRP content. Our results show a relationship between the diffuse gut endocrine system and feeding and nutrition in locusts. The ampullar endocrine cells are in three-way contact with the midgut luminal contents, with the primary urine from the Malpighian tubules and with the haemolymph. They are thus ideally positioned to play an integrative receptor-secretory function in the regulation of a variety of post-ingestive processes, such as enzyme secretion, absorption, gut motility or nutrient metabolism. PMID- 9866882 TI - Rostral-caudal variation in troponin T and parvalbumin correlates with differences in relaxation rates of cod axial muscle. AB - Relaxation rate is an important determinant of axial muscle power production during the oscillatory contractions of undulatory locomotion. Recently, significant differences have been reported in the relaxation rates of rostral versus caudal white muscle fibers of the Atlantic cod Gadus morhua L. The present study investigates the biochemical correlates of this rostral-caudal physiological variation. Using denaturing gel electrophoresis, a series of fresh muscle samples from the dorsal epaxial muscle region was analyzed and several differences were detected. First, a gradual shift occurs in the expression of two troponin T isoforms along the length of the body. Second, rostral muscles were found to contain significantly greater amounts of parvalbumin than caudal muscles. Third, two soluble Ca(2+)-binding proteins, in addition to parvalbumin, were also detected in the rostral muscle samples yet were absent from the caudal samples. This suite of rostral-caudal variations provides a strong biochemical basis for regional differences in the relaxation rates of cod white muscle. PMID- 9866883 TI - [Cancer pain: economic considerations and denied therapeutic chances]. PMID- 9866884 TI - [34th Annual Congress of the American Society of Clinical Oncology. 16-19 May 1998, Los Angeles, USA. Breast cancer (II)]. PMID- 9866885 TI - [Simultaneous infusion of vinorelbin and taxol as first-line chemotherapy in metastasized breast cancer]. AB - PURPOSE: To evaluate the activity and toxicity of simultaneous infusion of vinorelbine (VNB) and paclitaxel (T) as first line chemotherapy in advanced breast cancer patients (pts). PATIENTS AND METHODS: 33 pts with histologically proven advanced breast cancer were treated with VNB 25 mg/m2 and T 150 mg/m2, both drugs given by i.v. infusion over 3 hours, with cycles repeated every 3 weeks. Granulocyte-colony-stimulating factor (G-CSF), 300 micrograms subcutaneously, was given on days 7 to 12 of each cycle in the first 10 patients. RESULTS: From October 1995 to July 1996, 33 untreated pts entered the study. Characteristics of the pts were the following: median age 53 years (29-71); median WHO performance status 1 (0-3); pre/postmenopausal 8/25; prior adjuvant chemotherapy 16; prior radiotherapy 8; dominant disease sites: soft tissue in 6; bone in 7, viscera in 19; number of metastatic sites: 1 in 18, 2 in 9, 3 in 6 pts. In 31 evaluable pts we observed: 1 CR (3%) and 14 PR (45%), for an overall response rate of 48%. Median time to response was 2 months; median time to progression and median survival were 7 and 22+ months, respectively. Median number of cycles was 6. Myelosuppression was the dose-limiting toxicity, with G 4 neutropenia occurring in 22% of the pts and neutropenic fever in 6% of the pts. Other toxicities were generally mild with nausea in 52% of the pts; mucositis in 15%; constipation in 12%; peripheral neuropathy in 46.5%. Alopecia was universal. CONCLUSIONS: Simultaneous infusion of VNB and T is well tolerated and active in untreated patients with advanced breast cancer. Median survival (22+ months) is similar to that reported with anthracycline-containing regimens, although response rate appears to be lower. It is likely that higher response rates may be achieved with a higher dose-intensity. PMID- 9866886 TI - [Evaluation study of the opinions on the Di Bella multi-therapy (MDB) of tumor patients]. AB - PURPOSE: To know and to evaluate the opinion of cancer patients tumors who chose voluntarily to undergo the traditional anti-cancer treatment about Di Bella's therapy when the scientific debate was at its climax. MATERIALS AND METHODS: From March 1, 1998 to May 25, 1998 a questionnaire was distributed to 50 cancer patients. The questionnaire was anonymous, self-administered and included 9 questions. RESULTS: Overall, 35 (70%) patients were in favour of Di Bella's method.; 39 (78%) patients think that it is a successful step against cancer; 35 (70%) patients were willing to undergo the treatment. CONCLUSIONS: The main reasons for the complete trust in Di Bella's therapy were: mass media's power, Prof. Di Bella's image, the absence of toxicity in his therapy. These reasons will be reevaluated at the end of the trial. PMID- 9866887 TI - [Transcranial Doppler sonography in the pre- intra- and post-operative evaluation of 85 patients undergoing carotid endarterectomy]. AB - PURPOSE: To evaluate the usefulness of transcranial Doppler sonography during carotid endarterectomy. PATIENTS AND METHODS: Pre-, intra-, and postoperative transcranial Doppler was performed in 85 patients who underwent carotid endarterectomy. Intracranial collateral pathways were evaluated preoperatively and continuous monitoring of middle cerebral artery flow velocity was performed during carotid surgery. Transcranial Doppler was repeated in each patient 1 and 7 days thereafter. RESULTS: Values of residual middle cerebral artery flow velocity during preoperative digital compression of the common carotid artery were significantly correlated (r = 0.56; P < 0.001) with those obtained during cross clamping. Middle cerebral artery flow velocity was significantly lower during cross-clamping with respect to pre-clamping (P = 0.006), in patients with neurologic symptoms (P = 0.001), EEG alterations (P = 0.001), or defective collateral blood supply through the anterior communicating artery (P = 0.01). Postoperative flow velocity values of the MCA were significantly higher at day-1 examinations (P < 0.0001) with respect to preoperative values, and were no more significantly different at day-7. CONCLUSIONS: Transcranial Doppler is a useful method to evaluate the risk of cerebral ischemia during carotid endarterectomy cross-clamping, to identify those patients in which shunt insertion is mandatory, and to evaluate the postoperative effect of carotid endarterectomy on cerebral hemodynamics. PMID- 9866888 TI - [Effect of thermal mud baths on normal, dry and seborrheic skin]. AB - PURPOSE: To investigate the curative effects of thermal water and thermal muds in various cutaneous pathologies. PATIENTS AND METHODS: Modifications of phmetry and sebometry using sulphur thermal muds in normal, dry and seborrheic skin have been studied. The three groups of patients have been submitted to a 14 day treatment with thermal muds. RESULT: The application of thermal mud normalized the value of cutaneous pH and sebometry. CONCLUSIONS: These beneficial effects are long lasting in individuals who have a prolonged treatment with thermal muds. PMID- 9866889 TI - [Transdermal route as an alternative to oral administration of opioids in cancer pain]. AB - Fentanyl TTS, the only transdermal opioid, represents a real tool for a better quality of life in patients with cancer pain. In this paper we report a short description of the pharmacologic properties and administration procedures of this drug that is a useful alternative when other opioids recommended on the third step of the WHO analgesic ladder, are ineffective or present unbearable side effects (nausea and/or vomiting-severe mucosites and dysphagia). In particular we indicated some changes and adjustments switching from morphine per os to fentanyl TTS. In addition we report the results of a study carried out in our Pain Therapy Center on 49 patients with severe oncologic pain, previously treated with opioids and other drugs associations. Our results indicated a good control of continuous nociceptive cancer pain, with a better quality of life and lesser side effects to respect the previous regime of orally opioids. PMID- 9866890 TI - [On the subject of pancreatic ductal carcinoma]. AB - Pancreatic cancer is a dismal disease. The 5-year overall survival ranges from 1% to 5%. Surgery is the only curative treatment available for this cancer, but it is indicated only in selected patients with a less than 4 cm tumor. In these patients, survival rate is about 30%. We have considered several aspects: the very difficult early diagnosis, the correct diagnostic flow chart, actual surgical procedures and new trends in biologic and genetic research. It is likely that better results can be achieved by defining an "early pancreatic cancer" and establishing how to detect it. This could be the wrigth one way is to significantly improve the survival of these patients. PMID- 9866891 TI - [p53: prospects for gene therapy of cancer]. AB - PURPOSE: To evaluate the prospects for cancer gene therapy through restoration of wild-type p53 (wt-p53) protein expression and/or function. DESIGN: To review the most significant data reported in the literature with particular attention in dissecting the biological questions that are still open and need to be clarified to improve TP53-based gene therapy. RESULTS: Considerable experimental evidence obtained in vitro and in vivo indicates that wt-p53 protein can suppress the transformed phenotype of several types of cancer in humans as well as other species. Wt-p53 protein suppress transformation by inducing different biological effects including maintenance of genomic stability, inhibition of cell proliferation, induction of apoptosis, differentiation or senescence. All these findings have rendered p53 a potential helpful target for therapy of many types of human cancers. CONCLUSIONS: Different experimental strategies based on i) TP53 gene-replacement, ii) restoration of wt-p53 activity, iii) replication of defective lytic viruses specifically in altered p53-expressing tumors, have given promising results in vitro and, in some cases also in vivo. At present, a few phase one clinical trials have been started for some of the gene-replacement strategies. PMID- 9866892 TI - [Management of ventricular arrhythmia in chronic congestive heart failure]. AB - PURPOSE: To evaluate efficacy and indications of therapy for ventricular arrhythmias in patients with chronic congestive heart failure. DESIGN: A review of most significant and recent clinical trials was performed. RESULTS: In patients with severe left ventricular dysfunction, the desirable actions of antiarrhythmic drugs are attenuated and their negative inotropic and proarrhythmic actions are enhanced. Treatment should be limited to patients with malignant ventricular arrhythmias, or to patients considered at high risk. When indicated, amiodarone is usually well tolerated and safe. The prevention of sudden death in patients with heart failure should be based on optimized therapy of pump failure, reducing left ventricle work load and modulating neurohormonal systems with ACE-inhibitors and betablockers drugs. Further, an important role is held by anti-ischemic therapy, revascularization procedures, anticoagulant therapy and prevention of electrolytes unbalances. Patients with sustained of high risk arrhythmias, resuscitated from a cardiac arrest, should be considered for transvenous Implantable Cardioverter Defibrillator (ICD) implant. CONCLUSIONS: Ventricular arrhythmias are common in heart failure patients, represent an important cause of sudden death and the choice of treatment is difficult because of the complexity of underlying mechanisms, frequency of adverse reactions and the severity of left ventricular dysfunction. PMID- 9866893 TI - [A case of maternal and neonatal infection due to Listeria monocytogenes]. AB - A Listeria monocytogenes infection may develop during pregnancy by eating sausages, fresh meats and milk products derived from infected animals. According to the period in which the infection starts, the pregnancy outcome can be abortion or pre-term or at term delivery. The infection can pass from mother to fetus and can cause a serious neonatal sepsis. Listeriosis in pregnant women can be asymptomatic or may present as an influenza-like syndrome. This case report, along with other published cases, demonstrates how hard is to make a correct diagnosis of listeriosis during pregnancy. Since this is mainly related to the aspecificity of symptoms, it is very important to have a high suspicion and to take a careful patient history. PMID- 9866894 TI - [Epistemiologic reflexions on the discovery of I. P. Semmelweis]. PMID- 9866895 TI - Accreditation: IAPM-National Board for Laboratory Medicine. PMID- 9866896 TI - Seroepidemiology of ocular toxoplasmosis-profile of an urban population. AB - 200 uveitis cases and 100 controls were serologically analysed for Toxoplasma antibodies (Ab) using indirect fluorescent antibody test (IFAT-IgG, Igm) and enzyme linked immunosorbent assay (ELISA IgG, IgM). Ophthalmologically cases were segregated into 4 groups anterior uveitis, posterior uveitis, pan uveitis and varied presentation uveitis. Toxoplasma seropositivity of 32% in cases and 4% in controls was established. IHA, IFAT, ELISA detected 20%, 18% and 32% cases as seropositive respectively, IFAT being most specific (100%) and ELISA most sensitive (41.37%). Insignificant change in Ab titre was observed in sequential samples of seropositive cases. Posterior Uveitis cases had the maximum seropositivity (41.7%). Highest seropositivity was in 16-25 years age group with no sex preponderance. Dietary habits and occupational history had no bearing on Toxoplasma infection. Results indicate that serology in mandatory for diagnosing Toxoplasma as a cause of uveitis, 2 or more tests on a single serum sample detecting IgG and Igm Ab are the best indicators of infection. PMID- 9866897 TI - Rapid detection of enteric fever by coagglutination and countercurrent immunoelectrophoresis. AB - A total of 463 patients clinically suspected of enteric fever and 100 healthy individuals were investigated by coagglutination (COAG) and countercurrent immunoelectrophoresis (CIEP) for rapid diagnosis of enteric fever. The S. typhi was grown in blood culture in 32 (6.91%) and Widal test was positive in 126 (27.21%) cases. The serum COAG with local antiserum was positive in 165 (35.64%), serum COAG with standard antiserum in 163 (35.21%), Blood culture supernatant (BCS) COAG in 153 (33.05%), serum CIEP for antigen (Ag) in 118 (25.09%), BCS CIEP in 99 (21.39%) while serum CIEP for antibody (Ab) was positive in 34 (7.34%) cases. Only two healthy controls revealed positive COAG result with local antiserum. The sensitivity of all antigen detection tests was 100% except BCS CIEP (98.25%) in the first week of fever and declined rapidly to 75.79% for serum COAG tests, 69.47% for BCS COAG and dramatically to 37.89% for serum CIEP for Ag and 22.11% for BCS CIEP tests during the second week whereas the sensitivity of serum CIEP for Ab detection rose from 17.54% to 23.16% from first to second week of illness. In view of the resulting data, it is suggested that both COAG and CIEP may be employed for the rapid diagnosis of enteric fever in the routine clinical setup. PMID- 9866898 TI - Spectrum of breast disease in young females: a retrospective study of 1315 patients. AB - A total of 1315 breast lesions seen in women up to 40 years of age were registered in the files of Pathology Department. Out of these 807 were benign and 508 malignant lesions. Breast carcinoma was very rare up to second decade of life but its incidence progressively increased with age. Among benign tumours fibroadenomas were the commonest tumours. This study shows a higher incidence of inflammatory diseases and malignant tumours in young females as compared to the reports from the West. PMID- 9866899 TI - Cytodiagnosis of discharge per vaginum. AB - Simple, rapid, inexpensive methods such as pH determination, wet mount, KOH mount, amine test, Gram staining of vaginal discharge were undertaken prior to Pap stained smears of 158 patients of leucorrhoea. We were able to detect non specific vaginitis (44.30%), Trichomoniasis (16.45%), Candida vaginitis (9.49%), gonococcal vaginitis (0.63%) and senile vaginitis (12 cases). No specific pathology was shown in 26 cases. 1.69% of the cases were of cervical erosion and 3.79% cases suggestive of squamous malignancy. PMID- 9866900 TI - "EXPATH" an expert system for pathology for the diagnosis of jaundice. AB - Computers are increasingly playing an important role in the field of medicine. A computer aided expert system has been developed for the differential diagnosis of jaundice. In this article the major features and application of this modular software are discussed. PMID- 9866901 TI - Study of proteases and other enzymes of Vibrio cholerae 01 E2 Tor and 0139 serotypes isolated in Yavatmal (Maharashtra). AB - V. cholerae 01 E1 Tor isolated during Cholera epidemic of 1994 and V cholerae 0139 serotype isolated during 1993 epidemic were subjected to the study of proteases and other enzymes. Out of 26 strains of V. cholerae 01 studied, gelatinase and caseinase activity was seen in 100 and 69.23 percent strains respectively. All strains showed catalase and oxidase activity. Of the other enzymes studied 19.23, 65.38 and 57.69 percent strains were positive for DNAse, lipase and phosphatase respectively. None of the strains showed lecithinase activity. Similar findings were observed in 22 strains of V. cholerae 0139 except all strains were positive for phosphatase activity. Role of enzymes in virulence is suggested. PMID- 9866902 TI - A simple screening test for the detection of heterozygous beta thalassemia. AB - Beta-Thalassemia is prevalent in a high frequency of 5-15% in selected geographical areas and ethnic communities. There is need to screen large populations for the heterozygotes of beta-Thalassemia. We have evaluated NESTROFT (Naked eye single tube red cell osmotic fragility test) with 0.36% buffered saline in this study. Of the population screened, 42 cases were positive with NESTROFT, of which 40 were positive with the confirmatory test for beta Thalassemia. This implies a positive predictive value and specificity of 100%, negative predictive value of 83.3% and sensitivity of 95.2%. PMID- 9866903 TI - Comparison of darkground microscopy with serological tests in the diagnosis of leptospirosis with hepatorenal involvement--A preliminary study. AB - Darkground Microscopy (DGM) was initially done on Plasma in the Department of Microbiology, Kasturba Medical College, Manipal for 44 cases of suspected Leptospiral hepatorenal involvement. A total of 60 sera were collected from these patients and stored at -70 degrees C. Passiive haemagglutination test (PHA) was performed on 25 sera and latex agglutination test on 35 sera (Leptospira antigen Seiken) Leptospira was detected by Darkground Microscopy in 12 patients (27.27%). Serology was positive in 16 samples (36.36%) of which 7 (15.90%) by PHA and 9 by Latex agglutination (20.45%). Serological tests confirmed the findings of Darkground Microscopy. DGM was found to be simple and rapid and could be performed on all suspected patients. PMID- 9866904 TI - Barrett's esophagus. AB - Contrary to earlier definition of Barrett's oesophagus which referred to the presence of metaplastic columnar epithelium of either gastric or intestinal type, the new definition lays stress on presence of specialised columnar epithelium (SEC) with goblet cells. Hence this study was carried out to determine the frequency of specialised columnar epithelium in Indian population. 150 cases of dyspepsia underwent esophagogatro duodenoscopy with esophageal biopsy. Slides were stained with routine H and E stain and alcian blue-PAS stain. Histologically, esophagitis was found in 75 (50%) and columnar epithelium-gastric type in 63 (42%) cases only 4 (2.6%) cases showed presence of specialised columnar epithelium which were labelled as Barrett's esophagus, giving an incidence of 2.6% in this study. PMID- 9866905 TI - Diabetic foot infections: a study of microorganisms associated with the different Wagner grades. AB - Patients with diabetes foot infections were evaluated over a two year period to assess the bacteriological spectrum in the different Wagner's grades of foot wounds. Most of the diabetic foot wounds were found to be polymicrobic in nature with an average of 3.07 organisms isolated per case studied. Amongst a total of 775 clinical isolates, 71.09% were aerobic; whereas 28.91% were anaerobic pathogens. Gram-positive organisms like Staphylococcus spp. and Streptococcus spp. formed almost 50% of the clinical isolates in the first two grades and were reduced to less one-fourth of the total organisms in the last two grades. These was a significant increase in the gram-negative organisms and anaerobes in the last two grades. PMID- 9866906 TI - External clues to inner malformations. AB - A malformation is a morphological defect of an organ, or a larger region of the body resulting from an intrinsically abnormal developmental process. In this analysis of 1421 neonatal autopsies performed between 1984 and 1993, 243 (17.1%) cases showed malformations. The data was analysed to find external cues to internal malformations. Twenty three (85%) of the 27 neonates with various facial abnormalities had associated internal malformations in the form of cardiac (n = 11; 40%), renal (n = 7; 25%), or gastrointestinal (n = 3; 11%) abnormalities and diaphragmatic hernia (n = 2). Sixty seven neonates had neural abnormalities. These were associated with cardiac (n = 7; 10%), renal (n = 12; 18%) or gastrointestinal (n = 7; 10%) abnormalities; 43 cases, however, did not have any associated malformations. Renal malformations (n = 66) had highest association with skeletal abnormalities (n = 9; 22%). As many as 45(76%) of the 59 cases with cardiac abnormalities had no external anomalies. Five cases of hypoplastic lungs were seen, all associated with external malformations. PMID- 9866907 TI - Shigella and the fluorinated quinolones. AB - Between July 1996 to June 1997 a total of 531 Stool samples from cases of acute diarrhoea, dysentry, colitis admitted to Kasturba Medical College Hospital, Manipal were investigated. Shigella were isolated from 29 samples (5.46%). Out of them, Shigella flexneri were 16 (55.17%), Shigella boydii 8 (27.58%), Shigella dysentriae 3 (10.34%) and Shigella sonnei 2 (6.89%). Five strains of Shigella flexneri and one strain of Shigella dysentriae were found to show resistance to Nalidixic acid and the newer fluoroquinolones viz., Ciprofloxacin, Norfloxacin and Ofloxacin. The in vitro 'R' factor transfer study showed that the resistance was not transferable. PMID- 9866908 TI - Teratoma of the liver--a case report. AB - A case of primary teratoma of the liver in a 10 months old male child is being reported for its rarity. It is the first case report in Indian literature. PMID- 9866909 TI - Sporotrichosis in an unusual location--dermatologically and geographically. AB - We report the unusual fixed cutaneous variety of sporotrichosis in a patient hailing from Uttarkashi (Uttar Pradesh) in the north-western part of India. PMID- 9866910 TI - Peliosis of spleen presenting as splenic rupture with haemoperitoneum--a case report. AB - Peliosis of spleen is usually associated with peliosis of liver. Oval to round markedly dilated blood filled cystic spaces predominantly in parafollicular area is the characteristic feature of peliosis to differentiate it from congested sinuses. Exposure to viruses chemicals, contraceptive steroids or chronic debilitating disorders including malignancies are the various possible etiological factors. Though peliosis may remain asymptomatic, fatal intraperitoneal haemorrhage may occur due to its rupture. We report a case of peliosis of spleen presenting as splenic rupture with haemoperitoneum. PMID- 9866911 TI - Dyshormonogenetic goitre with clear cell change resembling parathyroid adenoma--a case report. AB - Dyshormonogenesis is a rare cause of goitre. It is manifestation of mutation in genes controlling the proteins involved in synthesis of thyroid hormones. Though there is wide spectrum of biochemical and clinical presentation, pathological features are similar in all defects. Nonencapsulated nodules with minimal regressive changes and lack of lymphocytic infiltration are important diagnostic features. Clear cell change as seen in this case is common and needs to be differentiated from parathyroid adenoma. Hypercellularity with cellular atypia should not lead to misdiagnosis of malignancy. PMID- 9866912 TI - Cytodiagnosis of Kimura's disease. AB - Kimura's Disease, a rare chronic inflammatory disorder with a benign course affecting the lymphoid tissue is reported hare in an 18 year old South Indian male who presented with bilateral solitary swelling over the mastoid region. Its fine needle aspiration cytology characterised by the Warthin-Finkeldey type giant cells against a background of a bimodal population of lymphocytes and eosinophils prompted the diagnosis of Kimura's disease at cytology itself. This was confirmed at histology. This is the third cytological report of its kind being reported in English literature. Literature with regard to the cytological appearances of Kimura's disease has been reviewed and the various differential diagnoses discussed. PMID- 9866913 TI - Hyalinizing trabecular adenoma--a case report with FNAC histologic, MIB-1 proliferative index and immunohistochemical findings. AB - Hyalinizing trabecular adenoma is a recently described benign thyroid tumor, almost exclusively occurring in females. The morphological features of this entity overlap with both papillary and medullary carcinoma to varying extent. This, in turn, creates a situation of serious diagnostic pitfall particularly for a false positive diagnosis of papillary carcinoma in fine needle aspiration (FNA) cytology. False consideration of medullary carcinoma is also possible by the unwary especially if staining for Congo red and/or immunostaining for calcitonin is not resorted to. At histologic level, the distinctive architectural pattern is however of great help and thus poses a much lesser danger of misdiagnosis. We relate here our experience in a recently encountered case of hyalinizing trabecular adenoma and describe detailed FNA cytologic and histologic findings along with immunohistochemical profile using a panel of eight monoclonal antibodies. The tumor proliferative potential has also been assessed using MIB-1 (Ki-67) immunostaining. The various pros and cons of diagnostic pitfalls are discussed. PMID- 9866914 TI - Primary squamous cell carcinoma of the breast in a young woman--a case report and review of literature. AB - A thirty-four years old woman presented with pure squamous cell carcinoma of the breast, a very rare occurrence in the younger age group. A simple mastectomy with axillary clearance was performed, followed by adjuvant radiotherapy & chemotherapy. The patient is doing well, with no evidence of recurrence thirty months (2 1/2 years after the treatment. Primary pure squamous cell carcinoma of the breast is a very rare disease. The exact histogenesis still remains obscure. There seems to be no specific prognostic correlation with the morphological characteristics of squamous cell carcinoma of the breast. The clinical behaviour of this tumor is also uncertain. Thus, further study is required to determine whether primary pure squamous cell carcinoma of the breast is to be treated as a separate entity or as the common type of breast cancer (i.e. adenocarcinoma or adenocarcinoma with squamous metaplasia.) PMID- 9866915 TI - Benign intramucosal melanocytic proliferation a marker for primary malignant melanoma of rectum. AB - Benign melanocytic proliferation in the mucosal glands of rectum is described in a 56 year old female with primary melanoma of rectum. PMID- 9866916 TI - Myospherulosis of maxillary sinus--a case report with review of literature. AB - Myospherulosis is a rare condition. The authors report a unique case of this entity arising in paranasal sinus. The review of literature with emphasis on pathogenesis is also discussed. PMID- 9866917 TI - Compatibility problems after intravenous immunoglobulin--a case report. AB - Serious incompatibility was noted in a patient diagnosed as acute Guillain Barre syndrome treated with intravenous immunoglobulin. Patient had positive direct and indirect antiglobulin test and the auto control was negative. There was no clinical signs of hemolysis. Patient's blood group was O D positive and cross matched several units of ABO compatible D positive and D negative blood. Only one unit was compatible. These findings suggest that the particular intravenous immunoglobulin contained a mixture of saline and immune antibodies having different specificity. As the number of patients getting treated with intravenous immunoglobulin is on the rise more and more compatibility problems should be anticipated and should be borne in mind during serological testing and evaluation. PMID- 9866918 TI - Accreditation of clinical laboratories: current status. PMID- 9866919 TI - Lacto-phenol cotton blue wet mount of faeces. PMID- 9866920 TI - 1998 Podiatric Practice Survey. Statistical results. PMID- 9866921 TI - [Clinical review of 74 cases with miliary tuberculosis]. AB - Seventy-four cases of miliary tuberculosis were studied retrospectively. The mean age of the patients was 45.3 years. Twenty-two patients suffered from another underlying diseases. Six were infected with human immunodeficiency virus. Twelve had been treated with corticosteroids. Fever was present in 97.3 per cent of patients. Elevation of serum alkaline phosphatase was found in 67.6 per cent of cases. The skin reaction to tuberculin was positive in 61.2 per cent. Nodular shadows were found in the chest X-ray in 98.6 per cent of cases. The nodules were smaller than 2 mm in diameter in 52.7 per cent of cases. Other findings were enlargement of mediastinal lymph node (17.6%), cavities (23.0%), pleural effusion (27.0%), and consolidation (35.1%). Sputum cultures and urine cultures were positive for Mycobacterium tuberculosis in 76.8 per cent and 58.6 per cent of cases respectively. The diagnosis was confirmed by histopathological findings in some cases. The rate of positive biopsies was 61.5 per cent by bone marrow aspiration, 83.3 per cent by lymph node biopsy, 100 per cent by liver and lung biopsy. Antituberculosis therapy was successful in most of the patients. Seven patients died of miliary tuberculosis, 4 of them had adult respiratory distress syndrome. PMID- 9866922 TI - [Lung cancer in patients with sequelae of tuberculosis]. AB - To clarify features of lung cancer in patients with tuberculosis sequelae, we analyzed data on 15 cases (5.1%) who were diagnosed with lung cancer before death among 294 deceased cases with tuberculosis sequelae at our hospital. There were 12 men and 3 women, with a mean age of 64 years. Most of the 15 patients had pulmonary dysfunction, and 4 had received home oxygen therapy. All 12 men had a history of smoking, and 10 of them had squamous cell carcinoma of the lung. There was no definite correlation between the locations of the tuberculosis lesion and those of lung cancer lesion on chest X-rays. Twelve patients had had thoracoplasty for tuberculosis, and in 6 of these patients the lung cancer occurred in the same lung. Lung cancer was apt to be diagnosed in an advanced stage. However, in patients who received home oxygen therapy, diagnosis had been made at an early stage because of the frequent chest X-ray follow-up. We conclude that lung cancer is an important complication in patients with tuberculosis sequelae, and early diagnosis of lung cancer by careful follow-up is essential in the care of cases with tuberculosis sequelae who have poor pulmonary function and/or systemic conditions. PMID- 9866923 TI - [Investigations into the significance of routine health examinations for tuberculosis in teachers based on the analysis of results of extraordinary health examinations]. AB - School teachers are regarded as one of the danger groups in contracting tuberculosis infection and are subjected to strict tuberculosis controls, since when they develop tuberculosis, many school children are exposed to infection to the disease. However, the recent decrease in the incidence of tuberculosis in Japan has led to disputes concerning the significance of routine mass health examinations for tuberculosis. In this study, the significance of routine health examinations for tuberculosis in teachers was investigated by the analysis of the results of extraordinary health examinations carried out for tuberculosis in teachers as the index cases. A total of 496 extraordinary health examinations were carried out by Nagoya City from 1975 to 1986 and by Aichi Prefecture from 1980 to 1995. In 49 instances of these examinations, teachers were regarded as index cases, which included 25 teachers of public primary, middle or high schools and 14 teachers of private schools, including private instructors for piano, painting or calligraphy, and teachers for supplementary education. The results of these examinations in both groups were compared, regarding the routes of notification, the disease status of the index cases, and the frequency and the scale of the infections of tuberculosis observed among contacts with the index cases. "Group infections of tuberculosis" was defined as instances the infection in which 20 or more cases were infected by the index case, "small scale group infection" as 5-19 infected cases, and "cases with infection" as 1-4 infected cases. The result obtained were as follows. 1. The response rates to routine health examinations were 99.9% in the teachers of public primary, middle or high schools, and about 20-30% in the teachers of private schools. 2. The proportion of the cases notefied by routine examinations were 68.0% in the former group and 21.4% in the latter group. The cases notefied before the onset of the symptoms in the former group was significantly more frequent than in the latter group. 3. In the former group, no far advanced cases were identified, whereas 2 (14.3%) far advanced cases were identified in the latter group. The cases with amount of tubercle bacilli in sputum exceed 3 on the Gaffky scale were 32.0% in the former group and 61.5% in the latter group. 4. One (4.0%) case of the "group infections of tuberculosis" was observed in the former group, and 2 (14.3%) cases in the latter group. "Small scale group infection" was observed in 4.0% of the former group and in 21.4% of the latter group, and "cases with infection" in 8.0% of the former group and in 35.7% of the latter group. The frequency and the scale of the infections of tuberculosis observed among contacts with the index cases was significantly smaller in the former group than in the latter group. In conclusion, routine health examinations for tuberculosis for teachers seems valuable for the early diagnosis of tuberculosis cases and for the prevension of the infection of tuberculosis in schools. PMID- 9866924 TI - [Two cases of pulmonary disease due to Mycobacterium szulgai]. AB - This paper describes with two patients with pulmonary disease due to Mycobacterium szulgai. The first patient was a 67-year-old man who consulted a doctor at the outpatient clinic of the Internal Medicine of our hospital, complaining with hemosputum. A chest X-ray showed an infiltrative shadow in the right upper lobe. A smear test of the sputum was negative but a culture was positive for mycobacteria. Second patient was a 37-year-old man who was admitted to our hospital, complaining with cough and fever. A chest X-ray showed an infiltrative shadow with cavity in the right upper lobe. A smear test was positive and culture was positive for mycobacteria. Cultured isolates of the two cases were indentified as M. szulgai. These two patients were treated with isoniazid, rifampicin and ethambutol daily. Their clinical symptoms improved and their sputum smears and cultures converted to negative for mycobacteria. PMID- 9866925 TI - [Mechanisms of induction and expression of anti-tuberculous immunity]. AB - The induction of anti-tuberculous immunity highly depends on the cytokines produced endogenously at the initial stage of immunization. Among several cytokines, IFN-gamma appears to be the most important to generate antigen specific Th1 type of protective T cells in mice. IL-12 and IL-18, which are produced by macrophages in response to virulent mycobacteria, are responsible for stimulating NK cells to produce IFN-gamma. Once antigen-specific Th1 cells are generated, Th1-dependent macrophage activation was effective in the elimination of infected bacteria through enhanced production of reactive oxygen intermediates and reactive nitrogen intermediates. In Listeria monocytogenes, one of the intracellular bacteria, listeriolysin O (LLO) appeared to be responsible for the induction of endogenous IFN-gamma from NK cells. The possible mechanisms operating in the induction and expression of anti-tuberculous immunity are discussed with special reference to cytokine responses. An application of LLO to the induction of protective immunity is also discussed. PMID- 9866926 TI - [Multidrug-resistant tuberculosis]. AB - Multidrug-resistant tuberculosis (MDR-TB) is defined as one that is resistant to both isoniazid and rifampicin regardless of its resistance to any other antituberculosis drugs. According to "Study on the incidence of drug resistance for new admissions" reported by TB Sanatorium Council in 1992 in Japan, no increase of incidence was observed in either the first treatment or re-treatment as compared with former reports. However, U.S.A. study indicates a significant increase of MDR-TB, which is supposed to have been caused by a primary drug resistance prevailed in an infected area, or an acquired (secondary) drug resistance due to incomplete and/or faulty treatment for active TB. Many incidences were also reported for mass nosocomial infection of MDR-TB with HIV patients. In spite of these serious issues in U.S.A., MDR-TB has not yet been a major concern in Japan, while Japan should work out countermeasures in advance with careful observation of its tend. One of the causes of mass nosocomial infection of MDR-TB observed in U.S.A. is reported due to a delayed treatment after long procedures of TB identification and susceptibility tests followed specimen sampling. Rapid tests of identification and susceptibility for TB, MDR TB in particular, are long expected. The introduction of recent molecular genetics technology will help to develop new rapid tests. While a relationship between drug resistance and TB gene is recently known to certain extent, total mechanism of TB resistance cannot be fully explained with only certain gene identified in the connection with drugs. Early treatment is critical for MDR-TB with HIV patient, as their prognosis is far worse than MDR-TB with non-HIV. Aside from HIV infection, very limited drugs are available for the treatment of MDR-TB. Drugs should be carefully selected based on the resistance patterns of each strain as well as its side effects anticipated. PMID- 9866927 TI - [Multidrug-resistant tuberculosis. 1. Drug susceptibility testing and its standardization]. AB - Drug susceptibility testing is one of the most difficult procedures to standardize in mycobacteriology laboratories. International standardization of the test is needed for comparative evaluation of controlled chemotherapeutic trials, for epidemiological surveys on the prevalence of drug resistance, and for guidance in the treatment of tuberculosis patients. In 1996, a new procedure for drug susceptibility testing was proposed by an ad hoc committee of the Japanese Society for Tuberculosis. The proposal is for a test with the proportion method using an Ogawa egg medium similar to those recommended by the WHO. Resistance is expressed as the percentage of colonies that grow on critical concentrations of the drugs, i.e. 0.2 microgram/ml for isoniazid, 40 micrograms/ml for rifampicin, 10 micrograms/ml for streptomycin and 2.5 micrograms/ml for ethambutol. Strains of tubercle bacilli which are grown on drug-containing media represents more than 1% of the number of colonies that develop on drug-free media are considered to be clinically resistant to that agent. The results are recorded as susceptible (S) or resistant (R) on the laboratory forms. The proportion method using agar- and liquid-based media is used worldwide. In the near future a test with agar- or liquid-based media will be discussed for standardization of the test in Japan, as described by the National Committee for Clinical Laboratory Standards of the United States. PMID- 9866928 TI - [Multidrug-resistant tuberculosis. 2. Mechanisms of drug-resistance in Mycobacterium tuberculosis--genetic mechanisms of drug-resistance]. AB - Multidrug-resistant Mycobacterium tuberculosis infection is now world wide health problem. However, according to the recent advances of molecular biological technics, some of the genetic mechanisms of drug-resistance of M. tuberculosis has been uncovered. Generally, drug-resistance of M. tuberculosis was caused by point mutations in chromosomal gene. In isoniazid (INH) resistant M. tuberculosis, mutations and genetic deletions in catalase-peroxidase gene (katG), inhA gene, or alkyl hydroperoxide reductase gene were reported. We also found that about 15% of INH-resistant M. tuberculosis isolates lacked katG gene, and these isolates showed highly resistance to INH with MIC > or = 64 micrograms/ml. On the other hand, mutations and other genetic alterations in RNA polymerase beta subunit gene (rpoB) were the major mechanisms of resistance to rifampicin (RFP) with high frequencies of 90% or more. Our evaluation of the relationship between RFP susceptibility and genetic alteration in rpoB gene also showed that 95% of RFP-resistant M. tuberculosis isolates involved genetic alterations in 69 bp core region of rpoB gene. Moreover, these genetic alterations in rpoB gene were suspected as the resistant mechanism to other rifamycin antituberculosis drugs, such as rifabutin and KRM-1648. In addition, it was reported that point mutations in 16S rRNA gene (rrs) and ribosomal protein S12 gene (rpsL) induced M. tuberculosis as streptomycin (SM) resistant phenotype. We analyzed genetic alternations in rpsL gene of clinically isolates of M. tuberculosis, about 60% of SM resistant isolates were shown point mutation in this gene ant they were all high SM-resistant with MIC > or = 256 micrograms/ml. Furthermore, nicotinamidase (pncA) gene, DNA gyrase A subunit (gyrA) gene, and embB gene were reported as the responsible gene to pyrazinamide-, quinolone- and ethambutol-resistance, respectively. Although all mechanisms of drug-resistance were still unclear, these informations are very useful and helpful for development of rapid diagnosis system of drug-resistant M. tuberculosis. PMID- 9866929 TI - [Multidrug-resistant tuberculosis. 3. Epidemiology of drug-resistant tuberculosis in Japan]. AB - In Japan, the frequency of drug-resistant tuberculosis has been investigated every 5 years since 1950s and increase of initial and acquired drug resistance has not been observed. However, the mathematical model analyse of time trend of prevalence of drug-resistant tuberculosis and frequency of initial drug resistance in Korea shows that there is little difference of infectivity and/or proportion of clinical breakdown between susceptible bacilli and resistant ones. The prognosis of isoniazid (INH) and rifampicin (RFP) resistant tuberculosis cases in Fukujuji Hospital was investigated. 367 cases including 50 initial drug resistant cases were analyzed with life table analysis. 50% of all cases and 70% of initial drug resistant cases became negative, 13% of all cases and 4% of initial drug resistant cases remained as positive, 37% of all cases and 27% of new cases died. Among cases who did not convert negative within one year, 41% of all cases and 34% of initial drug resistant cases died. The prognosis of INH and RFP resistant tuberculosis cases were still not satisfactory. PMID- 9866930 TI - [Multidrug-resistant tuberculosis. 4. Treatment and prognosis of multidrug resistant tuberculosis]. AB - We studied the clinical characteristics, treatment and prognosis of multidrug resistant pulmonary tuberculosis patients retrospectively. In this study, multidrug-resistant is defined as both resistant to 0.1 microgram/ml of INH and 50 micrograms/ml of RFP at least. From 1990 to 1997, out of 1841 culture positive pulmonary tuberculosis patients, 76 patients (4%) proved to be multidrug resistant (53 males, 23 females, age 18-84, 40 originally treated cases and 36 relapse cases). Most of cases revealed resistance to other drugs in addition to INH and RFP. The combination of anti-tuberculous drugs were complicated and changed repeatedly. The incidences of administration of drugs were as follows; TH 62%, EB 58%, PZA 58%, KM 33%, PAS 33%, SM 29%, CS 20%, EVM 14%, CPM 3%. New quinolones, for example OFLX/LVFX, CPFX and SPFX, were also used frequently (62%). Eight percent of patients were operated. Bacteriologically effective drugs that meant culture negative were TH (14%), PZA (12%), KM (12%), EB (12%), SM (5%), new quinolones (16%). 67% of originally treated cases and 43% of relapse cases became culture negative. Many cases were treated for a long period. 19% of originally treated cases and 33% of relapse cases were treated more than three years. 11% of patients were died of tuberculosis. Major prognostic factors were diabetes mellitus (17%), malignancies (10%), non-adherence (9%) and other complications. Because of no absolutely effective treatment, we have to choose a treatment according to each patient. Development of new treatment is crucial. PMID- 9866931 TI - [Multidrug-resistant tuberculosis. 5. Human immunodeficiency virus infection and multidrug-resistant tuberculosis]. AB - Outbreaks of multidrug-resistant tuberculosis (MDR-TB) among human immunodeficiency virus (HIV)-infected persons reported in the United States were very serious and the risks were increased by the delay of diagnosis, rapid progression from infection to active disease, inadequate therapy and poor tuberculosis (TB) control. Prevalence of drug-resistant TB among HIV-infected patients in Japan was studied. The results of drug susceptibility were collected through the nationwide working group for a survey of HIV-infected TB. Data of susceptibility for 39 cases were obtained. The isolates of two cases were resistant to isoniazid and rifampicin (including clinical failure of response), although no outbreak of MDR-TB was found in Japan. Case study of a patient who developed MDR-TB revealed that drug resistance might be selected by insufficient anti-TB therapy. The rate of resistance to any of the anti-TB drugs in HIV infected patients seemed to be high, although strictly evaluation was difficult due to no standardization for drug susceptibility testing. Of 9 cases with resistance to any of the anti-TB drugs, 8 had extrapulmonary TB including 5 cases of disseminated TB. In contrast thirteen of 30 cases without drug resistance had extrapulmonary TB. Since it has been reported that HIV infection is related to increased rates of drug resistance of TB bacilli, treatment with four-drug regimen should be started and sufficient courses of therapy are needed in HIV infected TB patients. PMID- 9866932 TI - [Multidrug-resistant tuberculosis. Additional comment: primary multidrug resistant tuberculosis--diagnosis and treatment]. AB - Primary drug resistance is defined as the presence of resistant strains of Mycobacterium tuberculosis in a patient with no history of prior anti tuberculosis chemotherapy. In Japan, a recent study shows that the prevalence of primary resistance has been stable for two decades and that primary multidrug resistant tuberculosis (MDR-TB) is rare, which suggests the effectiveness of tuberculosis control. We presented four cases of primary MDR-TB that we had experienced from 1984 to 1997, and discussed an issue about diagnosis and treatment of primary MDR-TB. Of the four patients, two young men received surgical resection, which has resulted in a favorable outcome. Of the other two patients, one responded to long-term chemotherapy with ethambutol, ofloxacin and enviomycin. There have been no recurrence so far in the three cases. The rest case died due to progression of tuberculosis. Two of the four patients had been in contact with relatives who had died of MDR-TB. In conclusion, all the tuberculosis patients should be suspected to be primary MDR-TB when they had a history of a contact with a tuberculosis patient in whom chemotherapy had not been successful, and once patients are diagnosed as MDR-TB, surgical intervention should be considered as an adjunctive treatment. To prevent the emergence of primary MDR-TB, it is important to treat MDR-TB patients appropriately and to implement the infection control program. PMID- 9866933 TI - [Video laparoscopic adrenalectomy]. AB - BACKGROUND: The authors describe the surgical technique for right and left adrenalectomy and the per- and postoperative course of the patients. METHODS: Twenty-seven laparoscopic adrenalectomies were performed between November 1992 and February 1996: there were 18 women and 9 men ranging in age from 31 to 70 years (mean 50.8 years); the surgical procedure was a lateral decubitus transperitoneal flank approach in 26 patients, and a retroperitoneal approach in one. RESULTS: Twelve right and 15 left glands were removed. Adrenal diseases were primary aldosteronism in 20 patients, non-functional adenoma in 3 patients, 2 Cushing's syndrome, adrenal cyst in one and incidentaloma in one. Mean adrenal gland size was 2.6 cm (range 0.5 to 8 cm). Five patients (18.5%), were converted to laparotomy, for dissection problems in 4 and failure to locate the left adrenal gland in one. The mean anesthesia time was 205.7 min and the mean surgical time 141.2 min. Surgical morbidity was one adrenal vein injury sectioned close to the vena cava: the hemorrhage was controlled by laparoscopic suturing without conversion. No mortality occurred and postoperative morbidity was one minor chest infection. The mean postoperative stay in hospital was 3.7 days (range 2.8) for the non converted patients. Laparoscopic adrenal gland removal is safe and offers fast recovery and short hospital stay. Laparoscopic adrenalectomy combines the advantages of both the conventional anterior and posterior approach. CONCLUSIONS: Personal experience of laparoscopic adrenal gland removal is described: a safe adrenal vascular control and gland dissection were performed and the length of operation and postoperative course carefully presented. PMID- 9866935 TI - [An update in the diagnosis and treatment of tumors of the hepatic portal]. AB - BACKGROUND: Cholangiocarcinoma affecting the cross of the bile ducts is a tumour which has, although treatments, a high rate of mortality. METHODS: Between January 1986 and January 1996 six patients affected by Klatskin's tumor were studied; three of them underwent operation and were treated with insertion of a T tube (Kerr), whereas the other three were managed endoscopically with trans tumoral stenting. RESULTS: The survival rate was about seven months for patients who underwent operation and fifteen months for patients managed with endoscopic stenting. CONCLUSIONS: After a review of the literature and according to personal experience, it is underlined that only an early diagnosis can achieve best results, since the latest knowledge about hepatic anatomy derived from transplants allows more extensive resections. PMID- 9866934 TI - [The hemodynamic effects of the in-vivo administration of insulin-like growth factor I]. AB - BACKGROUND: Recent studies have demonstrated that IGF-I has several biological activities that correlate with the GH axis, by acting as a cell protecting factor and a promoting compound in different tissues and organs. Our latest findings have demonstrated a potential application of IGF-I in the treatment of postischemic renal injury, which frequently appears after a kidney transplant. The beneficial effect of the renal postoperative recovery probably correlates with the regulation of the vascular tone, in which IGF-I plays a role with other cytokines. However, this rises the question whether IGF-I has any effect on the general hemodynamic status. This study was designed to underline the intraoperative hemodynamic effect of exogenous IGF-I in an experimental setting of renal transplantation in swine. METHODS: Twelve female swine underwent a left renal autotransplantation. At the reperfusion the animals were separated in two groups. Group one served as control. Group two received 400 micrograms of IGF-I (added to the flushing solution). The animals were kept under complete hemodynamic monitoring over the operation. RESULTS: Among the different parameters studied (mean arterial pressure, mean pulmonary arterial pressure, pulmonary wedge pressure, central venous pressure, cardiac output, oxygen extraction ratio, systemic vascular resistance, oxygen delivery and oxygen consumption), any statistically significant difference between group one and two were observed. CONCLUSIONS: While the clinical administration of IGF-I requires further studies, the in vivo administration of this peptide is apparently well tolerated, and does not cause any hemodynamic instability to the operation. PMID- 9866936 TI - [Palliative cholangiojejunostomy on the duct of hepatic segment III. Our experience]. AB - BACKGROUND: Aim of the paper is to prove that indications for cholangiojejunostomy, in cases when a mean and long term palliation may be expected, are better than those offered by the use of prostheses, without the well known and early bile duct complications. METHODS: Between 1984 and 1995, 12 patients with obstructive jaundice due to malignancy at the hepatic hilum were treated by segment III Roux-en-y or omega cholangiojejunostomy. In 1 patient with no communication between right and left lobe of the liver, biliary-enteric bypass to segment VI duct was also undertaken; in another with neoplastic stenosis of pylorus gastrojejunostomy was performed. Seven patients had carcinoma of the gallbladder, three hilar cholangiocarcinoma, one gastric carcinoma and one recurrent pancreatic carcinoma. RESULTS: There were two postoperative deaths, one for myocardial infarction and one for biliary peritonitis. Six patients had complications: four biliary fistulas with spontaneous resolution and two wound infections. Seven of the ten surviving patients experienced complete resolution of jaundice for at least 4 months. The mean survival of surviving patients was 9.7 months. Nine patients developed recurrent jaundice. CONCLUSIONS: Segment III cholangiojejunostomy is a good palliative treatment in most patients with malignant hilar obstruction. PMID- 9866937 TI - [Anthropometric measurements of the male inguinal canal]. AB - BACKGROUND: A problem in the use of a prosthesis in the treatment of inguinal hernia is the form and dimension the mesh should have. There are no precise indications in the international literature and manufactured pre-cut prostheses available are not always suitable for all patients. In this study some measurements within the male inguinal canal have been compared with the patients' physical characteristics, such as weight, height and body mass index, and with inguinal hernia too. The purpose was to look for a relation between these data, in order to establish the form and size of the ideal mesh prosthesis for every patient before operation. A "list" of different prosthesis sizes could help both the surgeon and the manufacturers. METHODS: The study was carried out on 150 male patients who underwent surgery for inguinal hernia. RESULTS: The analysis of the results does not demonstrate a relationship between the data examined. CONCLUSIONS: The conclusion is drawn that there could be two solutions to the problem of mesh size: one being the expensive prepacking of meshes of various sizes, and the cheaper one being to find out a method which, not considering the internal ring variability, will not influence the production of a universal mesh. PMID- 9866938 TI - [The imaging diagnosis of colovesical fistulae secondary to diverticulitis]. AB - BACKGROUND: The diagnostic procedures proposed in the evaluation of sigmoidovesical fistulas complicating diverticulitis are various and their effectiveness is still not well established. METHODS: Personal experience is based on 14 cases of colovesical fistulas secondary to sigmoid diverticulitis. Several diagnostic tools were employed: plain abdominal film (10 cases), large bowel enema (12), urography (3), cystography (2), sonography (4), and CT (5). The semeiotics of these fistulas were distinguished in direct, indirect, secondary, and related to the underlying disease. RESULTS: The fistulous tract itself was recognized in 100% of the cases with sonography, in 75% with enema, in 60% with CT, in 33% with urography, and in none with cystography. Vesical gas was visible in 100% of the cases with sonography and CT, and in 40% with plain radiographs. Diffusion of contrast medium was present in 91% of the cases with enema, in 60% with CT, and in 33% with urography. Focal thickening and/or irregularity of the bladder wall was evident with cystography and urography in 67% of the cases, with CT in 60%, with sonography in 50%, with enema in 8%. Diverticulosis/diverticulitis was recognizable in 100% of the cases with CT, in 91% with enema, in 25% with sonography. A paravesical abscess was recognizable in 40% of the cases with CT, in 25% with sonography, in 8% with enema. CONCLUSIONS: The radiourological procedures, though of limited use in our series, have a poor effectiveness. Large bowel enema and, specially, CT confirm as the method with greatest accuracy in the evaluation of these fistulas. The sonographic examination, according to personal preliminary experiences, is a valuable diagnostic alternative. Sonography and CT allow analysis of the perivisceral structures and, if compared with barium enema, provide a larger number of information on diverticulitis, which is essentially an extraluminal disease, and its complications. PMID- 9866939 TI - Metastatic spinal cord compression. Clinical remarks. AB - BACKGROUND: Metastatic compression of the spinal cord is a frequent occurrence throughout the evolution of neoplastic disease. Possible clinical-diagnostic strategies and therapeutic management of this pathology are discussed in terms of survival and quality of life. METHODS: The study includes 59 patients (40 males and 19 females, with an average age of 48.4 years) with metastatic spinal compression treated surgically in our centre (in some cases with stabilization of the spinal segment involved). RESULTS: In 40 cases the localization of the primary tumor was known when the patient was admitted. The segment involved was the dorsal one in 41 cases. The most frequent type of tumor was pulmonary carcinoma in males and breast carcinoma in females. Average survival was 5.3 months. Treatment integrated by stabilization improved the quality of life in comparison to laminectomy alone. Survival was also influenced by the histological type and site of the primary tumor. CONCLUSIONS: Surgical treatment not only prolongs survival but, above-all, guarantees a satisfactory quality of life. PMID- 9866940 TI - [Hepatic incidentaloma. A case report of cholangiocarcinoma]. AB - The finding of incidentalomas is becoming increasingly frequent during the course of routine diagnostic tests. At the hepatic level, the preclinical diagnosis of primary tumours has enabled a growing number of patients to undergo surgery with reduced operating trauma and greater oncological radicality. The authors report a case of cholangiocarcinoma whose pre-clinical diagnosis enabled its resection in spite of the patient's advanced age. PMID- 9866941 TI - [A rare case of bilateral spigelian hernia]. AB - A rare case of bilateral spigelian hernia is presented and the anatomy and clinical problems related to this disease are discussed. The role of TC scan in the diagnosis of cases with difficult clinical evaluations is stressed. PMID- 9866942 TI - [A strangulated perineal hernia. A rare case of intestinal obstruction]. AB - A rare case of intestinal obstruction due to strangulated perineal hernia, observed among 533 small bowel obstructions operated since January 1982 until December 1994 (0.2%) is described. Less than 100 cases are reported in literature. The etiologic, anatomical and clinical aspects of perineal hernia are examined. The reported case is of the primitive, congenital type caused by defect of the rectovaginal fascia fusion, in correspondence of the Douglas. Preoperative diagnosis is very difficult. Strangulation of this type of hernia is rare, the hernial sac surrounding tissue being elastic. In case of strangulation, symptoms of intestinal obstruction appear. In the reported case the preoperative diagnosis was "acute abdomen", because there was defence in hypogastrium and in the right iliac fossa. A straight abdominal radiography has not been performed. In the reported case the patient, a 22-year-old woman, was promptly operated and the incarcerated intestinal loop released. A regular postoperative period followed. The importance of a complete clinical examination, of a straight abdominal radiography and of a promptly performed operation is underlined. PMID- 9866943 TI - Acute Meckel's diverticulum in a senior patient. AB - Meckel's diverticulum is the most common congenital abnormality of the intestinal tract, occurring in 2% of autoptic studies. The case of an 85-year-old man referred to the Emergency Surgery Unit for intestinal obstruction and lower gastrointestinal tract bleeding is reported. Surgical exploration revealed a complicated Meckel's diverticulum full of coproliths, immersed in pus and blood. Examination of the resected diverticulum showed necrotic diverticulitis in the absence of ectopic gastric or pancreatic tissues. PMID- 9866944 TI - [The diagnostic contribution of computed tomography in a case of volvulus of the small intestine after an ileoanal anastomosis]. AB - A case of acquired volvulus as a cause of late small bowel obstruction, in a patient who had undergone a colectomy with ileal pouch-anal anastomosis two years previously, is described. Computed Tomography (CT) accurately demonstrated signs of adhesive strangulating obstruction, detecting also a radial disposition of the loops around the mesenteric root and an abnormal position of the superior mesenteric artery and vein on the right side of the aorta. From a literature survey, there are no previous studies reporting the CT appearance of a small intestine volvulus developed as a complication of an ileal pouch-anal anastomosis. PMID- 9866945 TI - [Cranial metastases from a follicular carcinoma of the thyroid. A clinical case report and review of the literature]. AB - Follicular carcinoma, in association with other prognostic factors, identifies a class of "high-risk" patients in which a more aggressive treatment is indicated. In fact, in patients operated on for follicular carcinoma bone metastases, also many years after primary surgery, are reported. A case of skull metastases in a young women occurred 3 years after a near total lobectomy for follicular thyroid carcinoma is described. After six years the patient underwent total thyroidectomy, resection of skull metastases, radiometabolic therapy with I131 180 m Ci; now she is disease free at three years of follow-up. The biology of metastases in thyroid follicular carcinoma, risk factors and diagnostic and therapeutic controversies are analyzed. Good prognostic factors are considered age < 45 years, good degree of differentiation of primary tumor, small size of bone metastases and early appearance after primary diagnosis. The conclusions is drawn that bone metastases are the worst prognostic factors. In case of single lesion, surgical treatment allows good results. PMID- 9866946 TI - [Pneumoretroperitoneum due to pneumatosis cystoides intestinalis. An etiopathogenetic and clinico-diagnostic assessment. A clinical case report]. AB - A case of pneumoretroperitoneum which came out to be caused by pneumatosis cystoides intestinalis, after careful and proper diagnostic evaluation, is described. Physiopathology, etiopathogenesis and clinical peculiarities of this infrequent pathology are examined; most useful tests to be performed in diagnostic differential evaluation, and clinical and surgical therapeutic approaches are also described, especially facing rare complications of PCI, such as pneumoperitoneum and pneumoretroperitoneum. PMID- 9866947 TI - [Breast carcinoma in men. The surgical treatment of a case under our observation]. AB - A case of male breast carcinoma in a very young subject (seventeen years old) is described. Breast carcinoma in male is infrequent, and at a so young age it is really rare. The patient was subjected to "Patey" mastectomy with removing the lymph nodes of the breast with the tumor, and subcutaneous mastectomy at the opposite side breast. As agreed with the oncologist other therapy was carried out. PMID- 9866948 TI - [Bilateral renal oncocytoma. A clinical case and review of the literature]. AB - Renal oncocytoma is an epithelial tumour of the kidney, originally considered benign neoplasm, but now it is thought have a limited but real malignant or metastatic potential. Usually monolateral, in 6% of cases it is bilateral in a synchronous or metachronous way. A case of bilateral renal oncocytoma in a 56 year-old woman, observed by chance during a follow-up carried out after breast cancer is described. The etiopathogenic and clinical-therapeutic aspects of this neoplasm are examined, and the conclusion is drawn that the most correct therapeutic approach must aim at both the protection of kidney function and complete removal of the tumour. PMID- 9866949 TI - [Ultrasonography of the uterine cervix in pregnancy. Curve of normality in a longitudinal and cross-sectional study]. AB - BACKGROUND: The aim of the study was to establish a normality curve of cervical length variations during pregnancy in our pregnant women population, to be compared with pathological cases (risk of preterm labor and cervical incompetence). DESIGN: A prospective longitudinal and cross-sectional study in women with single pregnancy and without risk factors for preterm labor was carried on. SETTING: Obstetric and Gynecology Department, University of Brescia, Italy. POPULATION OR SAMPLE: One hundred and thirty-four pregnant women with single pregnancy, 112 cross-sectionally and 22 longitudinally followed, with labor at term, were selected. Sixty-eight were nulliparous, 66 pluriparous. Multiple pregnancy, previous placental and vaginal bleeding were excluded. METHODS: A transvaginal probe was used to assess uterine cervix and the patients were studied from the 12th to 41st gestational week. Statistical analysis was carried out by Student's "t"-test and Z-test. MEASURES: Cervical longitudinal diameter, internal uterine os and funneling were assessed. RESULTS: No statistically significant differences emerged in the curves with regard to nulliparous and pluriparous patients. The cut-off between normality and pathology, from the 24th to the 32nd gestational week (2ndSD), seems to correspond to a cervical length lower than 25 mm; a significant progressive decrement of the cervical longitudinal diameter begins from the 28th gestational week. CONCLUSIONS: These results allows the gathering of reference data about normality to compare with pathology (cervical incompetence and preterm delivery), which could be useful both for prevention and follow-up of these cases. PMID- 9866950 TI - [Fatty acid composition of the cervical mucus obtained during ovulation and at the term of pregnancy]. AB - BACKGROUND: Aim of this work is to evaluate the different fatty acid composition of cervical mucus obtained during ovulation and at the term of pregnancy. METHODS: The fatty acid composition in cervical mucus was determined in 14 non pregnant women during expected ovulation (cervical score > 10) and in 12 at term pregnant women. Following extraction, (acidification and transesterification), the identification and quantification of fatty acids was performed by gas chromatographic analysis, with the aid of a specific software. RESULTS: In both groups of samples, palmitic acid, stearic acid and oleic acid were the prevalent acids comprising more than half of the total amounts. Compared to non pregnant samples, in pregnant cervical mucus, elevated levels of oleic acid were pointed out, while mean levels of miristic acid and stearic acid were lower. In the samples of cervical mucus drew at the term of pregnancy, arachidonic acid levels mean values were higher when the first period of labour was started. CONCLUSIONS: The pregnancy-induced biochemical changes in fatty acid pattern could likely be correlate to the variations of the physiochemical properties and to the physical appearance that cervical mucus undergoes during pregnancy. The elevated levels of arachidonic acid, during the first period of labour, may be correlated with prostaglandin production by intrauterine tissues (amniotic fluid, amnion, chorion, decidua, myometrium) and probably by cervical mucus. PMID- 9866952 TI - [Maternal attitude and psychophysical adaptation of women undergoing primary and secondary cesarean section]. AB - BACKGROUND: The increase of cesarean sections, which is typical of the most industrialised countries, resulted in a higher rate of repeat cesarean sections. The previous CS is the main indication for cesarean delivery. The purpose of this study is to compare women who had a primary cesarean with women who had a repeat cesarean section in order to assess their different emotional reactions as well as their psychophysical consequences. METHODS: A questionnaire was administered to 36 patients who had a primary cesarean section and to 34 patients who had a repeat cesarean section, 6-12 months after birth. All these women were similar for age and gestational age of the birth. RESULTS: Results underline that patients are more disposed to accept the operation in repeat cesarean rather than in primary cesarean. Women who have a repeat cesarean section are more likely to accept this kind of delivery since the beginning, with positive effects on their postoperative course. Women who have a repeat cesarean section face more serenely not only prenatal but also post-natal period and show less serious psychophysical sequelae with respect to primary cesarean section because of their previous experience. CONCLUSIONS: As a result, an appropriate psychological support coupled with adequate information can reduce discomfort in cesarean patients. PMID- 9866951 TI - Prenatal diagnosis using the triple test. AB - METHODS: A screening study performed on 2,803 pregnant women using the "triple test" is reported. RESULTS: Nine hundred and twenty-one had a high prior risk, having > 35 years while, after the screening, only 201 women had a positive test at risk higher than 1:270, and underwent to amniocentesis. The detection rate (DR) for all abnormalities was 91% while for Down's syndrome (DS) it was 87.5% and for neural tube defects 85.5%. Foetal abnormalities were detected in 20 cases (1:10) while 181 were false positive cases (6.5%), of which 151 for DS (5.4%). False negative were observed only in 2 cases within 2,339 at term pregnancies. CONCLUSIONS: The authors retain that high DR is related to the exactness of determination of gestation age calculated by scan and to the homogeneity of the examined population. PMID- 9866953 TI - [Biochemical markers of bone remodeling in the prevention of postmenopausal osteoporosis]. AB - Normal bone mass is one of the principal factors in the prevention of osteoporosis for women in advanced age. Therefore, it is necessary to have efficacious serum and urinary markers available to screen the bone development and the bone resorption. Even if we are also far from identifying an ideal marker, the measurement of some substances such as pyridinoline (Pyr) and deoxypyridinoline (D-Pyr) seems to be a first progress for efficacious and rather simple techniques. PMID- 9866954 TI - [Abnormal uterine hemorrhage in the climacteric. Diagnostic approach]. AB - Having made a detailed list of the invasive and non-invasive diagnostic techniques used to evaluate anomalous uterine bleeding in menopause, the authors conclude by affirming that hysteroscopy undoubtedly represents the most reliable diagnostic technique and that echography should be the preferred first diagnostic step since it allows the adnexa to be examined and is less invasive and cheaper. PMID- 9866955 TI - [Heparin-like factors in pregnancy. Report of a clinical case]. AB - Some authors have described the presence of circulating heparin-like anticoagulants; these substances are frequently associated with neoplastic pathology. Personal experience proves that this pathology of coagulation may be present also in pregnancy. The clinical case of a primipara, nullipara pregnant woman twenty-eight years aged is described; the case history showed that the patient presented circulating heparin-like substances. During this study, routine biochemical examinations were within normal limits as well as the coagulation tests. In the course of this experience, coagulation anomalies and hemorrhagic episodes in the intraoperative and postoperative period were not observed. PMID- 9866956 TI - Obstetric complications in Marfan's syndrome pregnancy. AB - Marfan syndrome is usually inherited as an autosomal dominant trait with high degree of penetrance. It is caused by an abnormal fibrillin gene located on chromosome 15q. Cardiovascular involvement in Marfan syndrome has been overstressed, although very little attention has been given to obstetric complications. Marfan syndrome may be responsible of cervical incompetence, abnormal placental site and post partum haemorrhagic complications. A 22-year-old woman with Marfan syndrome had mitral regurgitation since childhood. In addition aortic root dilatation was documented over six years by means of echocardiography and had been followed up regularly in a district hospital. Echocardiography six months before pregnancy had shown minimal mitral and aortic regurgitation and aortic root dilatation of 4.1 cm; left ventricular function was normal. Repeat echocardiography evaluations during pregnancy confirmed an aortic root dilatation. Routine booking and screening investigations were all within normal limits. At the 25th week, admission was necessary following a vaginal bleeding, without pain contraction. Echography showed a placenta praevia and cervical dilatation 2.8 cm of diameter. Bed rest and intravenous thocolitic therapy were immediately enhanced. A cervical cerclage, as described by McDonald, was placed. At the 37th week the patient was admitted and cerclage removed before the caesarean section. A healthy female of 2900 g was born. The postoperative period was favourable and patient was discharge after 7 days. In the present case, it is suggested that cervical incompetence and placenta praevia may be caused by an alteration of microfibrillar fibers. PMID- 9866958 TI - Sedation techniques in the dental treatment of patients with mental disabilities. PMID- 9866957 TI - [Therapeutic considerations in solitary cerebral metastases from uterine carcinoma]. AB - Brain metastases from endometrial carcinoma rarely involve the nervous system and are solitary in exceptional cases (< 1% of cases). Two cases of solitary cerebral metastasis from endometrial carcinoma are described. Two patients, submitted to the therapeutic protocol established for endometrial carcinoma, underwent surgery, radiotherapy and chemotherapy for solitary cerebral metastasis after at average interval of 18 months. Average survival was 46 months and death was due to progression of the systemic disease. An examination of our cases and those described in the literature has shown that, although these metastasis do not respond well to therapeutic treatment, a better outcome may be achieved by combined treatment consisting of surgery, radiotherapy and chemotherapy. PMID- 9866959 TI - [The evaluation of the postoperative response in surgery on the lower third molar]. AB - BACKGROUND: The impacted third molar is often excised in the absence of clear signs of disease. On this basis and taking also into account the usual surgical hazard, an analysis of the post-surgical behavior of 100 patients that had undergone excision of the impacted third molar, is made. METHODS: All patients were observed and treated by the same surgeon and were divided into two distinct groups based on the position and depth of the tooth in the bone as observed by orthopantomography. In the first group of patients, bone fenestration was carried out by the use of mallet and gouge and the tooth was excised without odontotomy. In contrast, in patients of the second group, odontotomy always preceded tooth excision. Patients of both groups were analyzed for several different parameters including age, sex, the reason for surgery, tooth position according to Pell and Gregory, the length of excision and, finally, swelling and the reduced mouth opening that occur after surgery. RESULTS AND CONCLUSIONS: Data reported indicate non-proportional results between the two groups; there is no apparent relation between post-surgical swelling and mouth opening, the latter being always present and very pronounced. PMID- 9866960 TI - Systemic corticosteroid therapy of oral vesiculoerosive diseases (OVED). An open trial. AB - BACKGROUND: We report the preliminary findings from our open study on how to optimize the doses and the response times to systemic corticosteroid therapy for three forms of oral vesiculoerosive diseases (OVED). METHODS: All twenty-one patients enrolled in the study were affected with OVED (10 OLP, 6 MMP, 5 EM) and received different doses of prednisone in relation to the type of disease and its severity. Daily mouthwashes with 0.12% chlorhexidine and application of miconazole gel were added to the therapy; its response time to treatment varied individually, the doses were reduced only after 75% of lesions had cleared. RESULTS: The average response time to treatment was 28 days. Clinical assessment showed that 95% of the patients had improved, whereas 71% had complete remission of oral signs and 48% experienced side effects. The degree of clinical resolution was: 70% complete remission after 26 days of therapy in patients with OLP, 33% complete remission after 38 days in patients with MMP, and 100% complete remission after ten days in patients with EM. CONCLUSIONS: Although a preliminary study, initial results showed different types and times of response to systemic corticosteroid treatment for the three diseases. Our findings strongly suggest that a unified therapeutic protocol for different OVED is not advisable, nor can systemic treatment be considered the first therapeutic approach. PMID- 9866961 TI - [Caries and malocclusions. A statistical-epidemiological study performed on 5399 children between 3 and 10 years old in the schools of Bari]. AB - BACKGROUND: The high incidence of decay and malocclusion is an undiscussed fact now. What is the relation between this incidence and sanitary-social standards is to define still. The aim of this survey was to evaluate the incidence of decay and malocclusions in relation to different sanitary-social standards. METHODS: The authors carried out an epidemiological-statistical study upon the incidence of decay and malocclusions on 5399 children whose age ranged from 3 to 10. The subjects belonged to some quarters of Bari: Palese, Santo Spirito, San Paolo e Poggiofranco. Some forms were filled in for any subject upon which data on oral health, oral hygiene and occlusal disorders were annotated. Then some groups was divided on subject's age and origin was created. RESULTS: The incidence of decay and malocclusions resulted extremely high in all considered groups. The examination of data showed a clanger gap about the oral hygienic standards between first three quarters, formed into A group, and Poggiofranco, in the last's favour. CONCLUSIONS: Students and their parents showed a serious sub estimation of the importance of oral health. An increase of quality and number of information on oral health is indispensable for many examination children and their respective parents. PMID- 9866962 TI - [The oral status in mentally retarded institutionalized patients]. AB - BACKGROUND AND METHODS: An epidemiological survey of dental and periodontal health status was conducted in a group of 263 institutionalized mentally retarded patients. RESULTS: The periodontal status in the study population was very poor. 66% of the subjects presented with a probing depth more than 3.5 mm (PSR = 3 or 4), 29% showed the presence of bleeding or calculus (PSR = 1 or 2) and only 5% showed a normal periodontal status (PSR = 0). No statistical relationship was found between periodontal status and age, sex, type or duration of psychiatric therapy. Concerning the dental status a total of 59 (26%) subjects was totally edentulous. A significant increase in the prevalence of edentulousness with increasing age was recorded. The incidence of edentulousness was higher among women than among men, while it was not significantly influenced by the type of psychiatric pathology, type or duration of psychiatric therapy. The mean number of remaining teeth in every subject was 12.9 +/- 9; the mean number of sound teeth was 10.8 +/- 8 and the mean number of decayed teeth was 2.1 +/- 3. The number of decayed teeth decreased with increasing age and it was influenced by the type of psychiatric pathology, but not by the type or duration of psychiatric pathology. CONCLUSIONS: Analysis of the results showed a very poor dental but especially periodontal status in mentally retarded subjects as compared to those found in studies performed in normal subjects. PMID- 9866963 TI - [The determination of the level of the external osteotomy line in Le Fort I interventions for pure maxillary advancement]. AB - BACKGROUND: Therapy of severe malocclusions needs a cooperation between orthodontist and maxillo-facial surgeon. The aim of this study is to evaluate how the ortho-surgical team can get a common base of programmation to transfer on split-casts osteotomic simulation reaching a more exact surgical result. METHODS: In this paper were considered patients operated by orthognathic surgery from January 1994 to June 1997 have been studied. ENVIRONMENT: Patients were operated in the Institute in which authors work. PATIENTS: Patients included were 120, 65 female and 55 male between 20 and 42 (mean 31). OPERATIONS: Patients were operated, for the most part, by combined operation at maxilla and mandible. RECORDINGS: The mean of values noticed was calculated adding all values and dividing for the number of values. RESULTS: The mean calculated is 33 mm (DS 2,22) of distance from canine cusp and 29 mm (DS 2,19) from mesio-vestibular cusp of the first upper molar. CONCLUSIONS: This study demonstrates that it is possible, with a statistically significant precision, to transfer osteotomic reference lines from clinical practice to a planning more pertinent to surgical performance. In this study we tried to standardize a value to increase precision of surgery, transferring easily feasible surgical measurements to split-casts. PMID- 9866964 TI - [The role of cell adhesion molecules in the formation of periapical granulomas]. AB - Cell adhesion molecules (CAMs) are cell surface proteins involved in the binding of cells, usually leukocytes, to each other, to endothelial cells, or to extracellular matrix. Specific signals produced in response to wounding and infection control the expression and activation of certain of these adhesion molecules. The interactions and responses then initiated by binding of these CAMs to their receptors/ligands play important roles in the mediation of the inflammatory and immune reactions that consult one line of the body's defense against these insults. Most of the CAMs characterized so far fall into three general families of proteins: the immunoglobulin (Ig) superfamily, the integrin family, or the selectine family. Recent studies have indicated that selectins (E,L,P) are implicated in cell trafficking, an important aspect of inflammation related process. Regulation of white blood cell trafficking from the blood vascular compartment to regions of pathogenic exposure is one of the most important functions of the immune system. The distinct phases of leukocyte migration include: rolling, activation, firm adhesion, transendothelial migration and subendothelial migration. The selectins have been implicated in the first step of this cascade. An inflammatory response is first evoked in the pulpal tissue in an attempt to neutralize the injurious agent and to dispose of damaged tissue and cells. The pulpal vessels dilate and blood flow to the tooth increases. At the same time, permeability of the vessels increases allowing leakage of fluid and leukocytes into the tissue. PMID- 9866966 TI - [The formation and persistence of a dental treatment phobia]. AB - The anxiety which accompanies dental care may in some cases represent a disabling handicap. The authors have outlined the characteristics of this type of dental anxiety and describe the typical pattern with which "odontophobia" is formed and persists over time. This enables them to understand the "attempted solutions" used to overcome this phobia, opening the way to the study of psychological interventions which may prove efficacious and effective. PMID- 9866965 TI - [Cell communication in the development of the oral cavity]. AB - The cadherins are a family of a calcium-dependent cell adhesion molecules that are regulated both spatially and temporally during development. Epithelial cadherin (E-cadherin) is present in the epithelial cells in both the embryo and yolk sac during organogenesis. In many studies, the presence of E-cadherin was analyzed during normal and disturbed craniofacial development with respect to palate and tooth formation. E-cadherin was present in epithelia of both ectodermal and endodermal origin. The expression level of E-cadherin increased with age and differentiation. The expression pattern of E-cadherin implies that this molecule has a role during normal development of the epithelia of the cranio facial complex. Morphogenesis and cell differentiation in the developing tooth are controlled by a series of reciprocal interactions between the epithelial and mesenchymal tissues. The exact molecular mechanisms operating in these interactions are unknown at present, but both structural components of the extracellular matrix (ECM) and diffusible growth factors have been suggested to be involved. In this review article we summarize our findings on the distribution patterns of three ECM molecules and two cell surface receptors during tooth morphogenesis through bud, cap, and bell stages of development. The examined molecules include fibronectina and tenascin, which all represent components of the mesenchymal ECM, the cell surface proteoglycan, syndecan, which functions as a receptor for interstitial matrix. Based on the observed changes in distribution patterns and on experimental evidence, roles are suggested for these molecules in epithelial-mesenchymal interactions during tooth development. C-Cam is a cell surface glycoprotein that is involved in cell adhesion and may play a role in histogenesis and organogenesis. It is a member of the carcinoembryonic antigen (CEA) gene family, which is a subfamily of the immunoglobulin gene superfamily. The presence of C-CAM in all proliferating craniofacial epithelia indicates that this molecule may play an important role in development. PMID- 9866967 TI - [A clinical case of primary malignant melanoma of the oral cavity]. AB - A rare case of primary malignant melanoma of the oral cavity is reported and a literature review of etiopathogenesis, biological bases and prognosis is made. The hypotheses to explain the very severe prognosis of malignant melanoma of the oral cavity with respect to that of skin are discussed. The therapeutic approaches are described as well as the frequent difficulty to perform them in relation to the anatomical region and the wide extension of the tumoral lesion. PMID- 9866968 TI - [The occlusal and functional rehabilitation of a lower canine in complete osseous inclusion]. AB - The aim of this paper is to show an occlusal and functional rehabilitation of a lower canine tooth in total osseous inclusion by means of surgical and orthodontic therapy. This rehabilitation is very important for mastication and for the correct function of the temporomandibular joint. PMID- 9866969 TI - Dentin dysplasia type I. Report of case and ultrastructural study. AB - Radicular dentin dysplasia (DD-I) is a rare hereditary dental alteration. It is characterized clinically by almost normal looking crowns and severe hypermobility of the teeth. The radiographic analysis, on the other hand, discloses the obliteration of all pulp chambers, the short, malformed roots and plenty of periapical bone radiolucencies on noncarious teeth. A case of radicular dentin dysplasia is presented. In this 43-year-old woman the diagnosis was supported, besides the clinical and radiographic analysis, by the pedigree of the proband, which showed the autosomal dominant pattern of feature transmission. Further more, the electron microscopic analysis of one extracted molar revealed the atubular structure of the secondary dentin, and its globular organization. PMID- 9866970 TI - [A biocompatibility study and the effects of slow-release antibiotic materials in the treatment of periodontal disease. I. The biocompatibility of cellulose acetate charged with 25% tetracycline hydrochloride. A clinical and scanning microscopic study of a case]. AB - A clinical and microscopical (SEM) investigation has been carried out on the biocompatibility of cellulose acetate fiber tetracycline with 25% of tetracycline hydrochloride (Actisite R). A subject with advanced periodontal disease was selected and a pocket of 8 mm of PD was chosen. A segment of fiber was inserted into the pocket for 8 days. After removal, PD and GI clinical parameters were detected and the fiber removed was analyzed at the scanning electronic microscope. The results showed clinical signs of inflammation after removal of fiber. SEM analysis showed macrophagic reaction, a typical sign of inflammatory response to material. The study suggests the need of more biocompatible materials, easier to use as delivery system of antibiotics in the treatment of periodontal disease. PMID- 9866971 TI - [A biocompatibility study and the effects of slow-release antibiotic materials in the treatment of periodontal disease. II. The biocompatibility and behavior of polyhydroxyethyl methacrylate (pHEMA) as a slow-release material for tetracycline of metronidazole. A study of 2 cases]. AB - The study analyses the possibility of using polyhydroxyethyl methacrylate as a material for the slow release of antibiotic in periodontal pockets. The antibiotics examined were tetracycline and metronidazole. The aim of the study was to evaluate the biocompatibility of the material with periodontal tissue and the efficacy of the 2 prepared systems. Two sites were selected in 2 periodontopathic patients who after non-surgical treatment presented pockets measuring 8 and 7 mm. A sheet of pHEMA containing tetracycline was inserted in one and in the other a sheet containing metronidazole: both were left for 8 days in the chosen pockets. At the start and end of treatment PD and GI clinical indices were measured and the DMDx microbiological test was performed to identify Aa of Pg and Pi. The tissue reaction to pHEMA was evaluated using SEM analysis of two samples collected after 8 days of treatment. The microscopic results showed the optimal biocompatibility of both samples. Differences were noted with regard to clinical and microbiological efficacy. It was observed that the sheet of pHEMA containing tetracycline resulted in the disappearance of bleeding and a reduced depth of survey. Moreover, microbiological results showed a significant reduction in Porphyromonas gingivalis. The sheet of pHEMA containing metronidazole showed a lower level of therapeutic efficacy. Although reduced depth was noted, gingival bleeding was persistent and the reduction of bacteria analysed was not significant. In conclusion, the authors confirm the optimal biocompatibility of the material and its easy application, although further research, especially for pHEMA with metronidazole, must be carried out to improve drug kinetics, trying to maintain an effective local concentration throughout treatment. PMID- 9866972 TI - [A case of parotidomegaly associated with eating disorders (anorexia/bulimia)]. AB - A case of parotidomegaly associated with anorexia is described, and the psychoneurological and etiopathogenetic theories of main feed disorders (anorexia/bulimia) are discussed. Particularly, a rare pathology of maxillofacial interest is presented: parotidomegaly associated with anorexia. The diagnosis and differential diagnosis of this pathology is discussed. PMID- 9866973 TI - [The construction of plates for intraoral registration with photopolymerizing resins. An original method]. AB - The recording of maxillo-mandibular relationships using Gothic arch assembly in dentulous subjects is useful not only in prosthetic rehabilitation, but also in the analysis of mandibular range of motion on patients with temporomandibular disorders. The traditional methodology to assemble Gothic arch plate is too long and elaborate, so it is remitted to the laboratory. This study describes a quick and simplified technique utilizing photopolymerizing resins which can be used by the dentist in his own office, in order to permit a greater use in the clinical practice. PMID- 9866974 TI - [The roles of apoptosis in lung injury and fibrosis]. AB - Apoptosis serves important roles in organ development, cell differentiation, and the maintenance of homeostasis. Lung injury studies have underlined the role of fibroblast and endothelial cell apoptosis during lung repair from acute lung injuries, and demonstrated apoptosis of alveolar epithelial cells in association with diffuse alveolar damage. Pulmonary fibrosis is characterized by the loss of lung epithelial cells and the proliferation of fibroblasts. It is possible that p53, p21, Fas, Fas ligand, and other apoptosis-regulating proteins play an important role in the pathophysiology of lung injury and fibrosis. PMID- 9866975 TI - [Thoracoscopic lung volume reduction surgery for emphysema]. AB - In recent years, several institutions have been performing lung volume reduction surgery (LVRS) for emphysema, and have reported that LVRS is effective for patients with emphysema that is resistant to other forms of therapy. However, questions remain about the relationship between the extent of resection and the therapeutic benefits. In our study, 15 men with emphysema underwent thoracoscopic LVRS. We analyzed the effects of LVRS on pulmonary function, exercise performance, and subjective symptoms after both bilateral and unilateral procedures. The patients who underwent the bilateral procedure, demonstrated significantly improved pulmonary function and exercise performance and relief of their subjective symptoms. Those who underwent the unilateral procedure, demonstrated significantly improved pulmonary function and relief of their subjective symptoms. Forced expiratory volume in 1.0 second increased by an average of 51% after the bilateral procedure, and 17% after the unilateral procedure. We conclude that thoracoscopic LVRS is an effective treatment for emphysema, especially with the bilateral procedure. PMID- 9866976 TI - [Clinical study of lung cancer complicated by pneumoconiosis]. AB - To elucidate the characteristics of lung cancer complicated by pneumoconiosis, we compared the clinical features of complicated and uncomplicated lung cancer cases. The average age at onset was higher for patients with pneumoconiosis than for those without. Because pneumoconiosis presents various respiratory symptoms, subjective symptoms were less important than objective symptoms in detecting lung cancer complicated by pneumoconiosis. Although fiberoptic bronchoscopy was the predominant diagnostic method, it could not be used with the pneumoconiosis patients. Diagnoses of lung cancer complicated by pneumoconiosis were made more after by percutaneous needle aspiration cytology. Squamous cell carcinoma is the most frequent form of cancer in patients with pneumoconiosis, which suggests that the inhalation of carcinogens may play a role in the pathogenesis of lung cancer. With respect to the smoking index (SI), histological specimens indicated that the incidence of squamous cell carcinoma in patients with pneumoconiosis was significantly higher in heavy smokers (SI > or = 600) than in light smokers (SI < 600) or nonsmokers. It is suspected that smoking affects carcinogenesis of pneumoconiosis. No differences in the distribution of lung cancer (right-left, hilar-peripheral) distinguished the 2 groups. Many of the patients with peripheral-type lung cancer and pneumoconiosis had tumors in the lower lung lobes. These findings underline the importance of encouraging pneumoconiosis patients to avoid lung cancer by not smoking, and to have lung cancer detected in its early stages by undergoing regular medical exams. PMID- 9866977 TI - [One year follow-up of pulmonary rehabilitation in patients with pulmonary emphysema--physiological outcome]. AB - To evaluate the long-term effects of pulmonary rehabilitation on physiological outcome, 12 patients with pulmonary emphysema were enrolled in an inpatient pulmonary rehabilitation program for 6 weeks. After discharge from the hospital, they were followed up for 1 year. The pulmonary rehabilitation program consisted of breathing retraining, thoracic mobilization, exercise training, and patient education. Although the subjects did not participate in outpatient maintenance group sessions after their discharge, they continued breathing retraining and exercise training at home. Their vital capacity improved significantly, and was sustained over the course of the year; other pulmonary functions, however, did not change significantly. Maximum exercise load increased 31% after the rehabilitation program; although it was 18% higher than baseline at follow-up one year later, that was not a significant change. The follow-up data on exercise traming had generally deteriorated 1 year after the rehabilitation program. The change in maximum exercise load from baseline before and after the inpatient pulmonary rehabilitation program correlated closely with the change in maximum exercise load thereafter to follow-up one year later (R = 0.62). We conclude that it is pessible to estimate long-term change in exercise capacity on the basis of short-term changes achieved during inpatient pulmonary rehabilitation. It may be necessary to develop maintenance programs of some kind to help pulmonary emphysema patients retain the benefits of pulmonary rehabilitation over the longer term. PMID- 9866978 TI - [Evaluation of serum KL-6 levels in summer-type hypersensitivity pneumonitis]. AB - A high level of serum KL-6 is a known feature of active pulmonary fibrosis. Some researchers have suggested that KL-6 is produced and secreted by type II pneumocytes. The present study evaluated serum KL-6 levels in patients with summer-type hypersensitivity pneumonitis (summer-type HP) (n = 6, 7 episodes), Mycoplasma pneumoniae pneumonia (n = 16), Chlamydia psittaci pneumonia (n = 3), Chlamydia pneumoniae pneumonia (n = 9), and bacterial pneumonia (n = 12). In addition, transbronchial lung biopsy (TBLB) specimens were examined pathologically in order to identify the site of production and secretion of KL-6. In patients with summer-type HP, the serum KL-6 levels exceeded 500 U/ml (2.996 +/- 2.016 U/ml), but was below 500 U/ml (302 +/- 126 U/ml, p < 0.001) in the patients with other infectious pneumonias, with the exception of two. One of these two patients with a high serum KL-6 level had adult respiratory distress syndrome due to Mycoplasma pneumoniae. The other had organizing pneumonia due to Chlamydia pneumoniae. TBLB specimens showed proliferative type II pneumocytes in all summer-type HP cases. We believe that the high serum KL-6 levels were produced by type II pneumocytes, and may provide a useful indicating serum marker for HP. Although serum LDH, serum CRP and PaO2 are known as monitoring markers in summer-type HP, our findings demonstrated no manifest correlations among these markers. However, serum KL-6 levels showed a strong positive correlation with serum LDH levels and an inverse correlation with serum CRP levels. These results suggest that serum KL-6 may be a better marker of the degree of disease activity than serum LDH, CRP, or PaO2 in summer-type HP. PMID- 9866979 TI - [Clinical evaluation of irinotecan combined with cisplatin by divided administration in patients with untreated primary non-small cell lung cancer]. AB - The efficacy and safety of irinotecan (CPT-11) combined with cisplatin (CDDP) were assessed in 24 previously untreated patients with primary non-small cell lung cancer. CPT-11 (60 mg/m2) and CDDP (30 mg/m2) were administered in combination at weekly intervals, on days 1.8, and 15 of the treatment course. During treatment, the patients were evaluated for adverse drug reactions and response. The response rate was 58.3% for all patients and 60.9% for the patients who completed the full treatment course. The median survival time was 13.0 months. The major adverse reactions were myelosuppression and gastro-intestinal disorders, but no treatment-related deaths were observed. Myelosuppressions included grade 3 or 4 leukopenia (25.0%) and anemia (33.3%). Grade 3 and higher gastro-intestinal reactions included nausea and vomiting (8.3%), diarrhea (12.5%), and anorexia (16.7%). These results suggest that combined weekly CPT-11 and CDDP therapy is capable of achieving a favorable tumor response with less toxicity, and thus worth consideration as a treatment option. Given that only 33.3% of the patients finished the full treatment course, further study should be devoted to the subject of CDDP and/or CPT-11 dosages. PMID- 9866980 TI - [Pulmonary edema associated with the Chinese medicine shosaikoto]. AB - A 45-year-old Japanese woman presented with a high fever, a nonproductive coughing, and severe dyspnea, and was admitted to another hospital. During the week prior to hospitalization, she had been given Shosaikoto for treatment of liver dysfunction of unknown etiology. Mycoplasma pneumonitis was initially suspected, so she was treated with antibiotics (clindamycin and minocycline) and received oxygen therapy. Pulmonary insufficiency worsened rapidly, and she was transferred to our hospital. On admission, a chest roentgenogram revealed bilateral alveolar infiltrates predominantly in the medial lung fields. Furosemide and high-dose methylprednisolone were immediately administered, but hypoxemia increased. When the PaO2 was 55.7 Torr while the patient breathed 100% oxygen, mechanical ventilation with positive end-expiratory pressure (PEEP) was started. Arterial blood-gas values improved dramatically, and the chest roentgenogram became clear. Our diagnosis of noncardiogenic pulmonary edema is based on the chest-roentgenographic findings, infiltration of inflammatory cells as seen in two lung-biopsy specimens and bronchoalveolar lavage fluid, the lack of findings of heart failure on physical examination and electrocardiography, and the good clinical response to PEEP. A positive lymphocyte stimulation test in response to Shosaikoto implicated this non-traditional herbal medicine as an etiologic factor in the non-cardiogenic pulmonary edema. Shosaikoto has been identified as the cause of interstitial pneumonia or eosinophilic pneumonia, but pulmonary edema associated with Shosaikoto has not been previously described. This case suggests that methylprednisolone treatment may be insufficient for Shosaikoto-induced pulmonary edema, and that mechanical ventilation with PEEP is very effective. PMID- 9866981 TI - [Allergic bronchopulmonary aspergillosis effectively treated with itraconazole]. AB - A 23-year-old man with bronchial asthma presented with fever, cough, and sputum. A chest X-ray examination showed pulmonary infiltrations in the left upper and lower lung fields with central bronchiectasis. Although his temperature came down with antibiotics, pulmonary infiltrations persisted with cough and sputum. Following bronchoscopy and an allergological examination, the patient was given a diagnosis of allergic bronchopulmonary aspergillosis (ABPA) based on Rosenberg's criteria, including peripheral blood eosinophilia, a high serum IgE level, immediate skin reaction to Aspergillus antigen, positive precipitating antibodies, and Aspergillus fumigatus in sputum. The patient was treated with itraconazole instead of corticosteroids. His respiratory symptoms, eosinophilia, and pulmonary infiltration then disappeared, and his IgE serum level gradually decreased. An antifungal agent alone was effective in treating this ABPA patient. PMID- 9866983 TI - [Peripheral primitive neuroectodermal tumor in parietal pleura]. AB - A 19-year-old woman was admitted to Kushiro city general hospital due to chest pain and dyspnea. Chest radiographs and computed tomographic scan showed a large intrathoracic mass adjacent to the pleura. Angiographs disclosed feeding veins of the tumor arising from lateral thoracic artery. Fine-needle aspiration cytology of the tumor revealed small round cells with a large nuclear/cytoplasmic ratio. Immunocytochemical study demonstrated that the tumor cells were positive for neuron-specific enolase and MIC 2 gene product. The diagnosis was primitive neuroectodermal tumor of the parietal pleura (also known as Askin tumor). Chemotherapy combined with peripheral blood stem cell transplantation reduced the size of the tumor significantly. However, multiple bone metastasis recurred, and the patient died 35 months after the start of therapy. PMID- 9866982 TI - [Hemophagocytic syndrome associated with tuberculosis and mycoplasma infection in two patients]. AB - Patient 1: A 48-year-old man was admitted to Osaka Red Cross Hospital because of fever and dyspnea. Laboratory examination revealed pancytopenia, liver dysfunction and hematostatic abnormality. Chest radiographs obtained on admission revealed ground-glass opacity in both lung fields, and an analysis of arterial blood showed severe hypoxemia (PaO2:46.8 Torr). Pulse therapy with methylprednisolone was started. Although the hypoxemia subsided and radiographic findings rapidly improved, pancytopenia persisted. Examination of bone marrow aspirate revealed mature histiocytes with marked hemophagocytosis. Amplified Mycobacterium tuberculosis direct tests of bronchoalveolar lavage fluid, sputum, urine, and bone marrow were all positive, and Mycobacterium tuberculosis was cultured from sputum and urine. Although the patient was taking antituberculous agents, his pancytopenia persosted. Treatment with etoposide induced remssion. Patient 2: A 19-year-old woman was admitted to Osaka Red Cross Hospital because of prolonged cough and fever. Laboratory examination revealed leukocytosis, liver dysfunction, and hematostatic abnormality. Serologic tests provided conclusive evidence of Mycoplasma infection and a CRP test was strongly positive. Chest radiographs obtained on admission revealed infiltration shadows in the middle and lower lung fields on both sides, with left pleural effusion. An analysis of arterial blood showed hypoxemia (PaO2: 54.2 Torr). Examination of bone marrow and pleural effusion samples revealed mature histiocytes with marked hemophagocytosis. Although treatment with antibiotics and pulse therapy with methylprednisolone was started, the patients respiratory functions deteriorated. Endotracheal intubation was performed. Therapy with etoposide induced remission. Hemophagocytic syndrome associated with Mycoplasma infection and tuberculosis appears to be exceedingly rare. In these 2 cases, it was difficult to achieve remission with therapy for the underlying infections, but etoposide treatment was effective. PMID- 9866984 TI - [Small cell lung cancer associated with subacute sensori-motor neuropathy in a patient whose symptoms subsided during chemotherapy]. AB - A 71-year-old man was referred to us with diplopia, left peripheral facial nerve dysfunction, ataxic gait and dysesthesia of the extremities. Neurological examination revealed mild reduction of sensation to pinprick and light touch in the left dominant lower leg. His standing position was wide based, and he showed Romberg's sign. The patient also presented signs of left peripheral facial, bilateral abducent, and left oculomotor nerve dysfunction. Serum levels of CEA, CA 19-9, and proGRP were high. 67Gallium scintigraphy showed an accumulation of radioactivity at the hilum of the right lung, and the findings of bronchofiberscopy were compatible with the diagnosis of small cell lung cancer. Because the symptoms gradually worsened to the point that the patient could not move by himself, chemotherapy and radiotherapy were initiated 3 months after the onset of symptoms. While under chemotherapy, symptoms of neuropathy subsided and the patient was able to walk with the aid of a walking stick. Although all symptoms were indicative of carcinomatous neuropathy, no antineuronal antibodies were detected in the patient's serum by immunohistochemical techniques. However, because the lung cancer deteriorated gradually despite therapy, the patient died of respiratory failure. At autopsy, tumor metastases were found in the pericardium, left lung, both adrenal glands, right hilum lymph nodes, and mediasternal lymph nodes. No microscopic signs of metastases were found in the frontal, parietal, temporal, or occipital lobes, or in the basal ganglia, thalamus, midbrain, pons, cerebellar vermis and hemispheres, or upper medulla. Histopathologically, there was no degeneration of neuronal cell bodies in cerebellar or cervical dorsal root ganglia; however, almost total loss of myelinated fibers or variegated demyelination of myelinated fibers was observed in the anterior, lateral and posterior funiculus at both cervical segments of the spinal cord. The number of myelinated fibers was smaller in the 5th and 6th cervical left ventral roots. The reason why the patient's symptoms subsided during chemotherapy was probably a suppression of antineuronal antigen by chemotherapy and the repair of myelinated fibers. PMID- 9866985 TI - [Severe pneumococcal pneumonia with acute respiratory failure and neutropenia]. AB - Severe pneumococcal pneumonia may precipitate both respiratory failure and neutropenia. The prognoses are considered to be very poor in such cases. We encountered three patients with pneumococcal pneumonia presenting respiratory failure and neutropenia, and successfully treated two. All three patients showed disseminated intravascular coagulation, and respiratory failure requiring mechanical ventilation. Neutropenia was treated with granulyte-colony stimulating factor (G-CSF), with methylpredonisolone administered by pulse therapy for 3 days to treat severe respiratory failure during the clinical course. All 3 patients were relieved of their respiratory failure at least for a few days following treatment. As a result of treatment with antibiotics and G-CSF, pneumococci disappeared from the patients' sputum. However, it has been reported that disintegration of the bacterial cell walls can release inflammatory components capable of causing secondary inflammatory reactions in the lung tissue. This suggested that the worsening respiratory failure experienced by our patients during their clinical course was a result of such reactions, and that methylpredonisolone was effective as the treatment of inflammation. PMID- 9866986 TI - [A mild case of acute eosinophilic pneumonia]. AB - We encountered a mild case of acute eosinophilic pneumonia in a 24-year-old man who developed symptoms once he started smoking. Bronchoalveolar lavage revealed eosinophilia. Chest computed tomography revealed a small amount of bilateral pleural effusion and multicentric panacinar shadows with peripheral ground-glass opacities. During the clinical course, the patient suffered only cough and fever, which subsided in 13 days without the administration of steroids. Although the clinical course in this case resembled that of simple pulmonary eosinophilia, we diagnosed it as a mild case of acute eosinophilic pneumonia because pleural effusion rarely accompanies simple pulmonary eosinophilia, symptoms developed when the patient began smoking, and interstitial involvement was suggested by bilateral effusion and ground-glass opacity. We look forward to the establishment of the disease concept of acute eosinophilic pneumonia, including mild cases, and a classification system for eosinophilic lung diseases, including acute eosinophilic pneumonia. PMID- 9866988 TI - [Marked airway constriction due to relapsing polychondritis]. AB - A 40-year-old woman was admitted to our hospital presenting with dry cough, sore throat, and fever. Her right auricle was markedly swollen and her ocular conjunctiva were reddened, suggesting scleritis. A chest computed tomogram showed narrowing of the trachea due to enlarged mediastinal soft tissues. These clinical findings suggested the patient had relapsing polychondritis. A cartilage biopsy from her right auricle was taken to confirm the diagnosis. Pathological findings revealed loosening and dissolution of cartilage and infiltration of lymphocytes, which were consistent with relapsing polychondritis. Although prednisolone was given to the patient, her symptoms were not alleviated. Methylprednisolone pulse therapy and oral diaminodiphenylsulfone were added to the patient's treatment regimen. With this combination, her symptoms gradually subsided. Constriction of the airway has been cited as one of the prognostic factors in relapsing polychondritis. In view of the danger of sudden death caused by airway obstruction, close observation of the patient in this case was considered necessary. PMID- 9866987 TI - [Pulmonary actinomycosis with "balls-in-a hole" appearance diagnosed by examination of bronchial lavage fluid]. AB - A 49-year-old man was referred to our hospital because of abnormal chest X-ray findings. Chest X-ray films showed infiltrative opacities in the right lung, and histological findings of a transbronchial biopsy specimen showed non-specific inflammation. The patient was treated with Ofloxacin for one month. After the treatment, chest X-ray films showed that the infiltrative opacities in the right upper lobe had decreased, but that opacities in the right lower lobe had increased, with an air meniscus sign. A chest computed tomography scan at the same time revealed that the remaining opacities contained multiple mass-like lesions within a cavity in the right S6, appearing as "balls in a hole". One year after the first visit, the patient visited the hospital again because of cough and sputum. A chest X-ray film showed that the size of the cavity in the right lower lobe had increased. The histological findings from a fresh transbronchial biopsy specimen revealed a non-specific inflammation again; however, black clots obtained from bronchial lavage fluid after biopsy were histologically identified as sulfur granules, a classic pathological indication of actinomycosis. This confirmed the diagnosis of pulmonary actinomycosis. The patient was treated with penicillin, and the opacities in the right lower lobe subsided. PMID- 9866989 TI - [Pulmonary veno-occlusive disease developed in a man given long-term treatment with anticancerous agents]. AB - A 67-old man was referred to our hospital because of dyspnea on exertion and severe hypoxia. He had been given, tegafur and OK 432 for seven years following an operation for gastric cancer. Pulmonary hypertension was noted by right heart catheterization. The findings of a transbronchial lung biopsy resulted in a diagnosis of pulmonary veno-occlusive disease. Pulmonary hemodynamic studies were performed for five different agents: nifedipine, beroprast sodium (PGI2), nitroglycerin, theophylline, and isosorbide dinitrate. However, none of these agents showed significant effects on pulmonary arterial pressure or pulmonary vascular resistance. Treatment with glucocorticoid relieved the patient's symptoms without any apparent effect on pulmonary hemodynamics. The long-term administration of anticancerous agents (tegafur) were thought to have caused pulmonary veno-occlusive disease to develop in this patient. PMID- 9866990 TI - [Clearance of gadolinium contrast agent by hemodialysis: in vitro and clinical studies]. AB - In order to evaluate the safety and pharmacokinetics of gadolinium contrast agents in patients with hemodialysis, in vitro and clinical studies were performed. Gd-DTPA and Gd-HP-DO3A solutions were dialysed by standard methods. One classic type (cellulose acetate) and two high-performance types (PMMA and polyacrylonitrile) were used. Eleven dialysis patients underwent normal dose contrast MRI followed by usual dose hemodialysis (HD). PMMA dialysis membranes were used for 8 patients and cellulose acetate membranes for 3 patients. Serum levels of gadolinium, BUN and creatinine were analyzed before and after the 1st HD and after the 2nd, 3rd and 4th HD. Side effects and edema were carefully observed. In vitro studies showed that 11.1 hours of dialysis would be necessary to remove 97% of the injected dose of contrast agent by using a first-order kinetic model of dialysis time. When dialysis membranes and contrast agents were compared, statistically significant differences were noted between dialysis membranes and all contrast agents. In the clinical study, 79.1% of the contrast agent was dialyzed after the 1st dialysis and 99.6% after the 4th dialysis. These results showed that all contrast agents and both dialysis membranes were suitable. Neither changes in laboratory parameters nor side effects were observed. The present study suggests, consequently, there are no contraindications when using the ordinary dose of contrast agent even in patients with dialysis. PMID- 9866991 TI - [Usefulness of CT-guided aspiration biopsy in combination with rapid cytology for diagnosis of benign pulmonary lesions]. AB - PURPOSE: To evaluate the diagnostic accuracy of computed tomography (CT)-guided aspiration biopsy in combination with rapid cytology for benign pulmonary lesions. MATERIALS AND METHODS: We performed percutaneous aspiration biopsy under CT guidance in combination with a rapid cytologic examination in 91 patients with pulmonary lesions. A 21-gauge modified Menghini needle coaxially placed through an 18-gauge needle was used in this procedure. Thirty-one lesions that were confirmed as a benign pulmonary lesion histologically, serologically, bacteriologically and/or clinically were evaluated in this study. RESULTS: In 28(90.3%) of 31 lesions, sufficient material for cytologic diagnosis was obtained from the aspiration biopsy. Specific benign diagnosis for benign disease was obtained in 13 lesions (41.9%), while nonspecific diagnosis for benign disease was obtained in 15 lesions (48.4%). The overall accuracy of the rapid cytological examination was 90.3%. Pneumothorax developed in 17 patients (54.8%), with 7 patients (22.6%) requiring chest tube drainage. Only one patient complained of mild hemoptysis, which subsided with hemostatic agents. CONCLUSION: Percutaneous aspiration biopsy combined with a rapid cytologic diagnosis provides a high degree of accuracy in the diagnosis of benign pulmonary lesions. PMID- 9866992 TI - [Combined scatter and attenuation correction for 201Tl myocardial perfusion SPECT using OS-EM algorithm]. AB - There are two possible ways to obtain scatter-corrected images with the ML-EM (maximum likelihood expectation maximization) algorithm: one is the subtraction of scatter estimate si from projection data pi, and then (pi-si) is used for scatter-corrected projection data (denoted as SC(T)); the other method is the addition of scatter estimate si to the projections calculated from the reconstructed image without performing data subtraction (SC(E)). This paper investigated these two ML-EM algorithms of combined scatter and attenuation correction on 201Tl myocardial perfusion SPECT imaging. Scatter windows were placed one full width at half maximum (FWHM) below and above the photopeak centerline. The scatter fraction in the primary peak was estimated using trapezoidal approximation by the triple energy window method. Phantom and clinical images were reconstructed using 6 iterations of ordered subsets EM algorithm (OS-EM). A cylindrical phantom with a cold-rod insert and a heart/thorax phantom with liver insert were used to evaluate scatter and the attenuation compensation technique. A cylindrical phantom filled with uniform 201Tl solution was used to evaluate statistical noise. The percent root-mean square uncertainty (%RMSU) was used as a quantitative measure of noise amplification. %RMSU showed that the SC(E) method amplified noise less in comparison with the SC(T) method, however, no significant difference in image quality was observed between the two methods. In conclusion, both the SC(T) and SC(E) methods provided significant and similar improvement in the removal of scatter in 201Tl myocardial perfusion SPECT imaging. PMID- 9866993 TI - [Basic study of MR-dacryocystography]. AB - We developed MR-dacryocystography as a non-invasive, safer imaging technique for canaliculi, the nasolacrimal duct and lacrimal sac by dropping saline solution and diluted Gd-DTPA solution into the eye. The diluted Gd-DTPA solution was found to create no local irritation in the eyes of rabbits and normal volunteers. Lacrimal sacs and ducts were well visualized in all of 10 normal volunteers by using the saline solution or the diluted Gd-DTPA solution. Canaliculi were visualized in 4-7 cases on thin-slice axial images. MR-dacryocystography was suggested to be a useful screening examination for lacrimal outflow disorders. PMID- 9866994 TI - [Easy graphical display of beam directions in three-dimensional converging radiation therapy: proposal for a radiation map]. AB - In stereotactic radiosurgery, non-coplanar isocentric beams are employed to concentrate the dose distribution on the planning target volume (PTV). However, the directions of incident beams must be determined with great care by using a digitally reconstructed beam's eye view (BEV) to prevent the irradiation of organs at risk. We present a new method of 2-dimensional graphical representation (radiation map) to facilitate the understanding of 3-dimensional relationships between incident beams and critical organs. After determining the isocenter and beam diameter, beam directions and critical organs are projected onto the imaginary sphere centered on the isocenter. The coordinate of the beam directions and the organs at risk can be expressed by latitude and longitude on the sphere. The contours of the organs at risk are displayed with a margin of the the radius of the radiation beam. Mirror images of the critical organs are also displayed to prevent irradiation by the opposing beams. The radiation map could be produced within 5 minutes using a workstation. Radiation maps, like DVH, will be very useful in the evaluation of radiation treatment planning. PMID- 9866995 TI - [Voice restoration and clinical outcome in patients undergoing laryngectomy with voice prostheses]. AB - Since the introduction of the tracheoesophageal fistula (TEF) technique for restoration of voice in 1983, we performed voice restoration using primary tracheoesophageal fistula in 83 patients undergoing laryngectomy. The success rate for voice restoration with TEF was 80.6% (good voice; 69.8%, fair voice; 10.8%). Thirty-one patients out of 51 patients who have been followed for over 3 years had 34 problems with TEF (15 granulations of TEF, 7 difficulties in insertion of voice prostheses, 4 stenoses of the tracheal stoma, 5 other reasons). However, most patients could use TEF by following appropriate procedures. Our results suggest that the Blom Singer TEF procedure is a safe, simple and effective means of alaryngeal communication. PMID- 9866996 TI - [The effects of high-dose steroid therapy on sudden deafness]. AB - The effect of high-dose steroid therapy on sudden deafness were investigated in 19 cases. A daily intravenous administration dose of hydrocortisone sodium succinate was tapered from 800 mg to 200 mg as follows; 800 mg 800 mg, 600 mg, 600 mg, 400 mg, 400 mg, 300 mg, and 200 mg. Another 19 cases, the control group, were treated with intravenous application of prednisolone, tapered from 50 mg to 10 mg in 10-mg steps and each dose applied for 3 days. There were no significant differences in the effect on hearing between the two groups as measured by three different analyses: criteria proposed by the Committee on Sudden Deafness of the Japan Health Ministry; magnitudes of improvement of mean thresholds at five frequencies from 250 Hz to 4 kHz in affected side; and the ratio of the magnitudes of improvement of mean thresholds in affected side to magnitudes of difference between the initial mean thresholds in affected side and those in contralateral side. These findings suggest that high-dose steroid therapy for sudden deafness is not so effective as it is for idiopathic facial palsy. This may be due to the difference in the mechanisms of development between the two conditions. No critical side effects were observed in our study, which would argue for the safety of our method of high-dose steroid therapy for routine clinical use. It might be advisable to conduct additional clinical studies to determine the effect of high-dose steroid therapy for sudden deafness, because our study was conducted on a small number of patients and was not double-blind or randomized. PMID- 9866997 TI - [Epidemiologic study of Chlamydia pneumoniae with ELISA]. AB - Chlamydia Pneumoniae (C. pneumoniae) is a bacterium involved in infections of the upper airway as well as in lower respiratory diseases. It has been reported that infections from this bacterium are prevalent worldwide and that the proportion of the population having the antibody is high. It is, however, difficult to identify the pathogen by routine bacterial examination because it is an obligate cytozoic bacterium. In addition, the examination has been feasible in only a limited number of laboratories because the determination of the serum antibody titer has required preparation of the antigen as well as sophisticated skill. We conducted an epidemiological survey of 320 healthy males and females in their 20s living in the three cities of Osaka, Kobe and Oita using "HITAZYME C. Pneumoniae", which is a recently developed kit for determination of anti-C. pneumoniae specific antibody. The statistical method used was the chi square test for the comparison of proportions. Mean proportion of the population showing positive antibody test was 58.1%. IgA antibody was positive in 42.8% and IgG in 46.5% of the above population. There were no statistically significant differences between districts of between genders in the percentages of cases positive for these antibodies. The results were comparable to those previously reported suggesting that C. pneumoniae is prevalent all over Japan. This kit was found to be an easy way to use the ELISA method and therefore to be clinically useful. PMID- 9866998 TI - [An assessment of physical and psychological stress of patients with facial paralysis]. AB - The purpose of this study was to determine if there is a relationship between the appearance of facial paralysis and physical or psychological stress. Subjects were 55 patients (23 men, 32 women) with facial paralysis (32 with Bell's palsy, 23 with Ramsay Hunt syndrome) who visited the Department of Otolaryngology, Nihon University Itabashi Hospital, from August 1994 to March 1996. At the time of the initial visit, patients were asked if they had any physical or psychological stress one week before the occurrence of facial paralysis. The degree of psychological stress was rated by Niina's Psychological Stress Response Scale 50 Items Revised (PSRS-50R). Answers were obtained from 52 of the 55 subjects. Forty patients (76.9%) reported that they had felt physically fatigued, suggesting some type of relationship between the occurrence of facial paralysis and physical stress. Twenty-seven patients (51.9%) reported the existence of psychological stress. However, the evaluation of psychological stress by PSRS-50R did not reveal a high level of psychological stress in any patient, indicating lack of a strong relationship between the occurrence of facial paralysis and psychological stress. Stepwise regression analyses were made to study the relationship between the degree of paralysis at the time of the initial visit and factors which affect the prognosis of the paralysis. Those patients who reported physical fatigue before the appearance of facial paralysis had more abnormal values on the nerve excitability test than those who did not report such fatigue. PMID- 9866999 TI - [Clinical significance of electrically evoked auditory brainstem response]. AB - Electrically evoked auditory brainstem responses (EABR) were recorded in 31 postlingually deafened adults, who had recently received cochlear implants (mini system, Cochlear Ltd). The wave consisted of three distinct positive peaks labeled P1, P2, and P3 with latency of 1.35 (+/- 0.14), 2.17 (+/- 0.18) and 4.08 (+/- 0.31) ms, respectively. The P3 threshold (EABR-T) and slope (EABR-S) were 0.9 (+/- 0.47) mA and 0.6 (+/- 0.28) muv/mA, respectively. The relationships between the EABR parameters (EABR T and -S of the P3 wave) and age, duration of deafness, promontory test and subjective response (T and C-level) were investigated. The scattergram showed a strong negative linear relationship between EABR-S and subjective T-level. This finding suggests that EABR-S is good measure of postoperative perception. PMID- 9867000 TI - [Distortion product otoacoustic emissions for the assessment of auditory sensitivity]. AB - Distortion product otoacoustic emissions (DPOAEs) were evaluated in 494 normal and 506 cochlear-impaired human ears, to determine whether DPOAEs depend on factors such as background noise, the shape of the pure tone audiogram, sex and aging, and whether a DPOAE test can perform well in distinguishing normal-hearing from hearing-impaired ears. The amplitudes of DPOAEs were measured at the frequency of 2f1-f2 (f1 < f2, f2/f1 = 1.22, f2 at 1, 2 and 4 kHz) using as stimuli two pure tones at level of 70 dB from an ILO92 Otoacoustic Emission Analyzer. The correlation coefficients between the DPOAE level and the auditory threshold decreased as the background noise levels at 1 kHz and 2 kHz increased. Therefore, it appeared that ears with large background noise levels would be inadequate for the study of DPOAEs predicting the hearing state. The sensitivity (normal-hearing ears identified as normal hearing) and the specificity (hearing impaired ears identified as hearing impaired) at the equal-sensitivity specificity condition were 80.7-86.7% at 1, 2 and 4 kHz, and the areas under the receiver operating characteristic (ROC) curves, which were used to estimate the test performance, were 0.88 for 1 kHz, 0.91 for 2 kHz and 0.92 for 4 kHz. Since these results suggest that a DPOAE can be used as a reliable technique for objective auditory tests, it is thought that actual values (DPOAE level: 4.3 dB at 1 kHz, 5.0 dB at 2 kHz and 2.9 dB at 4 kHz) of false-positive (hearing impaired ears identified as normal hearing) rates corresponding to 5% can be used in clinical evaluation to separate normal hearing from hearing-impaired ears. There was, however, a significant age effect at 4 kHz on DPOAEs in the ears with the same pure tone hearing thresholds, and the areas of the ROC curves in subjects ranging from 10 to 29 years old were larger than in subjects over 50 years (1 kHz: 0.88 to 0.94 versus 0.83 to 0.84, 2 kHz: 0.95 versus 0.89, 4 kHz: 0.95 to 0.96 versus 0.88 to 0.89). Therefore, it is thought that age-adjusted norms may be necessary for the accurate interpretation of DPOAE results. PMID- 9867001 TI - [Regional variations of the cytokeratin expression along the guinea pig cochlear turn]. AB - Expression of cytokeratin (Ck) proteins in the guinea pig cochlea was examined using a recently developed immunohistochemical method for celloidin section. This method has allowed us to observe a whole cochlea with excellent morphological preservation in a visual field under the light microscope. Anti-Ck antibodies FIL 7 (AE-1, Signet Lab.) and NCL-CK19 (Novocastra Lab.) stained all supporting cells of the organ of Corti, in which pillar cells, Deiters' cells, Hensen's cells, and Boettcher's cells showed a strong reaction. The histochemical labeling of pillar and Deiter's cells showed a gradual increment from the base to the apex. The immunohistochemically stained area in Hensen's cells increased toward the apex, which was attributed mainly to the increase in both the number and size of Hensen's cells from the base to the apex. In an electron-microscopic examination, changes in histochemical labeling of Hensen's cells did not appear from the base to the apex. Close examination revealed that reaction precipitates were distributed along a filament structure which seemed to correspond to the cytokeratin filaments. These results suggest that the Ck filaments give elasticity and flexibility to the supporting cells in the organ of Corti and provide mechanical strength to the organ of Corti at the upper turns, which vibrate with larger amplitudes, and that the Ck filaments in the supporting cell must be of considerable importance as a tuning mechanism. Immunohistochemical labeling for Ck was also observed in the spiral prominence cells, outer sulcus cells, interdental cells, and Reissner's membrane. The positively stained area in the outer sulcus increased from the apex to the base. PMID- 9867002 TI - [Application of clinical tests to routine medical practice]. PMID- 9867003 TI - [Concept of standard value and range in clinical tests]. PMID- 9867004 TI - [With special reference to clinical specimen handlings]. PMID- 9867005 TI - [Appropriate use of clinical tests in health-insurance managed medical care]. PMID- 9867006 TI - [New clinical tests that physicians should know for diagnosis of respiratory system disease]. PMID- 9867007 TI - [New clinical tests that physicians should know for diagnosis of cardiovascular disease]. PMID- 9867008 TI - [New clinical tests that physicians should know for diagnosis of digestive system diseases]. PMID- 9867009 TI - [New clinical tests that physicians should know for diagnosis of urological diseases]. PMID- 9867010 TI - [New clinical tests that physicians should know for diagnosis of hematologic diseases]. PMID- 9867011 TI - [New clinical tests that physicians should know for diagnosis of endocrine and metabolic disorders]. PMID- 9867012 TI - [New clinical tests that physicians should know for diagnosis of collagen diseases]. PMID- 9867013 TI - [New clinical tests that physicians should know for diagnosis of neuromuscular diseases]. PMID- 9867014 TI - [New clinical tests that physicians should know for diagnosis of infectious diseases]. PMID- 9867015 TI - [Clinical gene diagnosis]. PMID- 9867016 TI - [Effective usage of clinical tests (discussion)]. PMID- 9867017 TI - [Memorandum of a pathologist in the Japanese Red Cross Atomic Bomb Hospital]. PMID- 9867018 TI - [A case of isocyanate-induced hypersensitivity pneumonitis]. PMID- 9867019 TI - [A case of diffuse pulmonary metastasis of malignant pleural mesothelioma with elevated CYFRA in serum and pleural effusion]. PMID- 9867020 TI - [Case of diabetic ketoacidosis due to subacute thyroiditis in NIDDM associated with Basedow's disease]. PMID- 9867021 TI - [Case of protein-losing enteropathies due to gastrointestinal amyloidosis secondary to early-stage rheumatoid arthritis]. PMID- 9867022 TI - [A case of proscillaridin poisoning]. PMID- 9867023 TI - [Autoimmune diseases and retroviruses]. PMID- 9867024 TI - [Sleep apnea and non-invasive positive-pressure respiration]. PMID- 9867025 TI - [Recent topics on diagnosis and classification of diabetes mellitus]. PMID- 9867026 TI - [Expression of COX-1 and COX-2 and gastrointestinal diseases]. PMID- 9867027 TI - [Significance and procedures of evidence-based medicine]. PMID- 9867028 TI - Parental role models, gender and educational choice. AB - Parental role models are often put forward as an explanation for the choice of gender-atypical educational routes. This paper aims to test such explanations by examining the impact of family background variables like parental education and occupation, on choice of educational programme at upper secondary school. Using a sample of around 73,000 Swedish teenagers born between 1972 and 1976, girls' and boys' gender-atypical as well as gender-typical educational choices are analysed by means of logistic regression. Parents working or educated within a specific field increase the probability that a child will make a similar choice of educational programme at upper secondary school. This same-sector effect appeared to be somewhat stronger for fathers and sons, while no such same-sex influence was confirmed for girls. No evidence was found that, in addition to a same-sector effect, it matters whether parents' occupations represent gender-traditional or non-traditional models. Parents of the service classes or highly educated parents -expected to be the most gender egalitarian in attitudes and behaviours--have a positive influence upon children's choice of gender-atypical education. PMID- 9867029 TI - A telling tale: a case of vigilantism and its aftermath in an English town. AB - This paper considers one 'vigilante' episode in an English town in 1993 and its subsequent appearances in the press and in local 'crime-talk'. In so doing it a) proposes as an alternative to most current constructions of 'fear of crime' an interpretive approach grounded in place; b) considers the intersections between the generic 'law and order' preoccupations of the national press and the salience in local knowledge of a particular sequence of events (and their consequences for their dramatis personae); c) raises conjecturally some preconditions favouring the adoption of the 'vigilante' option amongst available styles of security seeking action. Theoretically, the paper demonstrates the relevance of locally circulating stories of crime and low-level street disorder to the contemporary understanding of crime, place and community. PMID- 9867030 TI - Photophysical properties of tin ethyl etiopurpurin I (SnET2) and tin octaethylbenzochlorin (SnOEBC) in solution and bound to albumin. AB - The photophysical characteristics of two second-generation PDT photosensitizers, tin ethyl etiopurpurin I (SnET2) and tin octaethylbenzochlorin (SnOEBC), have been measured in homogeneous solution and when bound to bovine serum albumin (BSA). The ground state and triplet state absorption spectra have been characterized, as have triplet lifetimes and quantum yields for intersystem crossing, singlet oxygen formation and photobleaching. In total, these parameters provide a complete set of data that can be used to quantitatively compare the photosensitizing efficiencies of these molecules. The photo-bleaching quantum yield of SnET2 is increased dramatically when it is bound to BSA, thus limiting the production of singlet oxygen at incident fluences above 1 J/cm2. In contrast, the quantum yield of photobleaching of SnOEBC is at least an order of magnitude lower than that of SnET2 under these conditions and does not significantly limit the photosensitization process for typical in vivo or in vitro fluences. This difference is expected to play a significant role in determining the relative photo-sensitizing ability of these compounds in vivo. PMID- 9867031 TI - Lethal consequences of simulated solar radiation on the nematode Caenorhabditis elegans in the presence and absence of photosensitizers. AB - The partial destruction of the earth's protective ozone layer has raised concerns about the impact of increased UV radiation on the earth's biological systems. In this study, polychromatic light sources were employed to observe the biological responses of the soil nematode Caenorhabditis elegans to simulated solar UV. Using various filter combinations, action spectra were constructed that approximated those generated previously with mono-chromatic radiation. In both cases, a mutant strain (rad-3) progressively lost its hypersensitivity as shorter wave-lengths were filtered out. In addition, both wild type and radiation sensitive (rad) mutants were irradiated with several combinations of filtered light sources in the presence and absence of two exogenous photosensitizers (ethidium bromide and bromodeoxyuridine). Treatment with either of the introduced photosensitizers increased photosensitivity to solar UV. Solar UV also induced a fluence-dependent reduction in fertility in wild-type animals. These experiments extend previous data and substantially expand our understanding of the biological responses of C. elegans to solar radiation. PMID- 9867032 TI - Identification of a single phosphorylation site within octopus rhodopsin. AB - Light-dependent phosphorylation of rhodopsin (Rho) is a first step in the desensitization of the signaling state of the receptor during vertebrate and invertebrate visual transduction. We found that only 358Ser of the photoactivated octopus Rho (oRho*) was phosphorylated by octopus rhodopsin kinase (oRK). Tryptic truncation of the C-terminal PPQGY repeats of oRho that follow the phosphorylation region did not influence spectral or G-protein activation properties of oRho but abolished phosphorylation. Despite significant structural differences between oRK and mammalian RK, these results provide further evidence of the importance of singly phosphorylated species of Rho* in the generation of arrestin binding sites. PMID- 9867033 TI - Kinetics of photobleaching of protoporphyrin IX in the skin of nude mice exposed to different fluence rates of red light. AB - The purpose of the present study was to determine the kinetics and the fluence rate dependency of the photobleaching of protoporphyrin IX (PpIX) in normal skin of Balb/c nude mice after systemic and topical application of 5-aminolevulinic acid (ALA). ALA was administered systemically (200 mg/kg body weight, i.p.) and topically (20% w/w ALA cream) to the mice. Fluences of up to 40 J/cm2 were delivered by a dye laser (636 nm) at fluence rates of 37.5, 75, 150, 300 and 500 mW/cm2. The photo-bleaching rate was constant within this range of fluence rates. This result suggests that there is no oxygen effect for PpIX photobleaching in this region for the skin of Balb/c nude mice. During light exposure the fluorescence decay followed neither first- nor second-order kinetics. The decay rate was slightly faster after systemic application than after topical application of ALA, but did not depend on the time (1-8 h) between application and analysis. PMID- 9867034 TI - The effects of photodynamic therapy on human neutrophil migration using bacteriochlorin a. AB - Bacteriochlorin a photodynamic therapy (BCA-PDT) caused inhibition of interleukin (IL)-8-activated neutrophil migration, at concentrations that did not induce membrane damage. Random migration and migration induced by other chemoattractants were also inhibited, indicating that the effect of BCA-PDT was not at the level of chemoattractant-receptor interaction but down stream. The BCA-PDT completely blocked superoxide production of phorbol ester-stimulated neutrophils indicating that superoxide production by neutrophils present in the tumor before and during BCA-PDT is not the cause of inactivation of tumor cells. Both type I and type II quenchers prevented inhibition by BCA-PDT but only in electroporated cells. Confocal laser scanning microscopy showed that the fluorescence of BCA was located inside the cell. These results show that the effects of BCA-PDT are intracellular and of a mixed type I/type II character and that the neutrophils present in the tumor during illumination probably do not contribute to tumor eradication by releasing reactive oxygen species. PMID- 9867035 TI - Birefringence measurement of rapid structural changes during collagen denaturation. AB - Linear birefringence, an optical property that results from a material's structure and composition, can be used to study dynamic changes in tissue structure. Single, 200 microseconds-long pulses from a Ho:YAG laser emitting 2.1 microns radiation were used to induce changes in the linear birefringence of rat tail tendon. Such changes were measured on a millisecond timescale. The measured rate coefficients describing the denaturation are not predicted by previous studies of collagen denaturation induced by slower, lower-temperature heating. Two types of laser-induced collagen denaturation can be differentiated: thermal denaturation, which appears rate-limited, and thermomechanical denaturation, which is observed at higher laser radiant exposures. Neither process is described by standard Arrhenius-type kinetic models. PMID- 9867036 TI - Recombinant phytochrome of the moss Ceratodon purpureus: heterologous expression and kinetic analysis of Pr-->Pfr conversion. AB - The phytochrome-encoding gene Cerpu;PHY;2 (CP2) of the moss Ceratodon purpureus was heterologously expressed in Saccharomyces cerevisiae as a polyhistidine tagged apoprotein and assembled with phytochromobilin (P phi B) and phycocyanobilin (PCB). Nickel-affinity chromatography yielded a protein fraction containing approximately 80% phytochrome. The holoproteins showed photoreversibility with both chromophores. Difference spectra gave maxima at 644/716 nm (red-absorbing phytochrome [Pr]/far-red-absorbing phytochrome [Pfr]) for the PCB adduct, and 659/724 nm for the P phi B-adduct, the latter in close agreement with values for phytochrome extracted from Ceratodon itself, implying that P phi B is the native chromophore in this moss species. Immunoblots stained with the antiphytochrome antibody APC1 showed that the recombinant phytochrome had the same molecular size as phytochrome from Ceratodon extracts. Further, the mobility of recombinant CP2 holophytochrome on native size-exclusion chromatography was similar to that of native oat phytochrome, implying that CP2 forms a dimer. Kinetics of absorbance changes during the Pr-->Pfr photoconversion of the PCB adduct, monitored between 620 and 740 nm in the microsecond range, revealed the rapid formation of a red-shifted intermediate (I700), decaying with a time constant of approximately 110 microseconds. This is similar to the behavior of phytochromes from higher plants when assembled with the same chromophore. When following the formation of the Pfr state, two major processes were identified (with time constants of 3 and 18 ms) that are followed by slow reactions in the range of 166 ms and 8 s, respectively, albeit with very small amplitudes. PMID- 9867037 TI - Physiologic responses to primary blast. AB - BACKGROUND: Primary blast injuries are produced by the blast shock wave. The critical determinant of survival is pulmonary injury, but acute cardiorespiratory responses to blast exposure are not well understood. The aim of this study was to investigate these changes. METHODS: Twenty anesthetized rats were exposed to moderate blast overpressure, 10 animals receiving thoracic and 10 receiving abdominal exposure. Another 9 animals acted as controls. Respiration, heart rate, and blood pressure were recorded continuously before, during, and for 6 hours after blast exposure. RESULTS: All animals exposed to thoracic blast demonstrated apnea, bradycardia, and hypotension after blast exposure, followed by a return to preblast values. No significant cardiovascular or respiratory changes were seen in animals in the other groups. CONCLUSION: Moderate thoracic blast injury produces a reflex triad of apnea, bradycardia, and hypotension that is not present after abdominal blast. These observations may have important implications for the immediate management of patients with blast injuries. PMID- 9867038 TI - Determinants of myocardial performance after blunt chest trauma. AB - OBJECTIVE: This study investigates whether factors that determine myocardial performance (preload, afterload, heart rate, and contractility) are altered after isolated unilateral pulmonary contusion. METHODS: Catheters were placed in the carotid arteries, left ventricles, and pulmonary arteries of anesthetized, ventilated (FiO2=0.5) pigs (31.2+/-0.6 kg; n=26). A unilateral, blunt injury to the right chest was delivered with a captive bolt gun (n=17) followed by tube thoracostomy. To control for anesthesia and instrumentation at FiO2 of 0.5, one group received tube thoracostomy only (sham injury; n=6). To control for effects of hypoxia without chest injury, an additional sham-injury group (n=3) was ventilated with FiO2 of 0.12. To generate cardiac function (i.e., Starling) curves, lactated Ringer's solution was administered in three bolus infusions at serial time points; the slope of stroke index versus ventricular filling pressure defines cardiac contractility. RESULTS: By 4 hours after pulmonary contusion, pulmonary vascular resistance, airway resistance, and dead space ventilation were increased, whereas PaO2 (72+/-6 mm Hg at FiO2=0.5) and dynamic compliance were decreased (all p < 0.05). Despite profound lung injury, arterial blood pressure, heart rate, cardiac filling pressures, and output remained within the normal range, which is inconsistent with direct myocardial contusion. The slope of pulmonary capillary wedge pressure versus left ventricular end-diastolic pressure (LVEDP) regression was reduced by more than 50% from baseline (p < 0.05), but there was no significant change in the slope of the central venous pressure versus LVEDP regression. By 4 hours after contusion, the slope of the stroke index versus LVEDP curve was reduced by more than 80% from baseline (p < 0.05). By the same time after sham injury with FiO2 of 0.12 (PaO2 < 50 mm Hg), the regression had decayed a similar amount, but there was no change in the slope after sham injury with FiO2 of 0.5 (PaO2 > 200 mm Hg). CONCLUSION: After right side pulmonary contusion, the most often used estimate of cardiac preload (pulmonary capillary wedge pressure) does not accurately estimate LVEDP, probably because of changes in the pulmonary circulation or mechanics. Central venous pressure is a better estimate of filling pressure, at least in these conditions, probably because it is not directly influenced by the pulmonary dysfunction. Also, ventricular performance can be impaired by depressed myocardial contractility and increased right ventricular afterload even with normal left ventricular afterload and preload. It is thus conceivable that occult myocardial dysfunction after pulmonary contusion could have a role in the progression to cardiorespiratory failure even without direct cardiac contusion. PMID- 9867039 TI - Blunt vascular injuries of the head and neck: is heparinization necessary? AB - BACKGROUND: Blunt vascular injuries to the head and neck (BHVI) represent some of the most devastating and morbid injuries seen by a trauma surgeon. This series reviewed the experience of a single institution to determine if diagnostic and therapeutic guidelines can be established for these uncommon injuries. In particular, the utility of anticoagulation in the treatment of these injuries is examined. METHODS: The institutional trauma registry of a single state-designated Level I trauma center was examined for patients with BHVI. Patients were identified and their charts reviewed individually with regard to multiple data points including the type of injury, its presentation, the treatment of the injury, and the functional outcome of the patient. RESULTS: Twenty-nine BHVI in 23 patients were reviewed from 1989 to 1997. No mortalities were noted. Among the injuries noted were 14 internal carotid artery dissections and 8 carotid artery tears. Thirteen patients had accompanying closed head injuries. Ten patients were diagnosed after an abnormal neurologic examination, and eight others were diagnosed after having carotid canal fractures. Heparin was started within 48 hours of injury in 4 patients (17%) and was used in a total of 12 patients (52%). No patient worsened neurologically after diagnosis independent of the use of heparin. Thirteen patients (57%) had no or minimal deficits upon discharge. CONCLUSION: BHVI represent a serious cause of morbidity in the patient with multiple injuries. Patients with closed head injuries and carotid canal fractures appear most at risk. A multicenter, randomized trial involving antiplatelet therapy, full systemic anticoagulation, or observation with a long-term functional assessment is indicated to determine the optimal management of these injuries. PMID- 9867041 TI - The epidemiology of thoracic aortic injuries in pedestrians. AB - BACKGROUND: Traumatic rupture of the thoracic aorta is recognized as a common cause of occupant death after rapid deceleration in motor vehicle collisions. The incidence of thoracic aorta rupture in pedestrian victims of vehicular collisions, however, is unknown. This study focuses on the epidemiology of injury to the thoracic aorta in pedestrian vehicular collisions. METHODS: We performed a retrospective analysis of all pedestrian fatalities and survivors of rupture of the thoracic aorta during a 6-year period at a regional Level I trauma center and medical examiner's office. RESULTS: There were 220 pedestrian fatalities during the study period. Laceration of the thoracic aorta was noted in 28 of the 220 pedestrian victims (12.7%). Two additional pedestrians survived laceration of the thoracic aorta, for a mortality of 94%. Hospital mortality was 66% (4 of 6). The comparative hospital mortality for patients with rupture of the thoracic aorta secondary to motor vehicle collision was 42%. CONCLUSION: The incidence of thoracic aortic injury in pedestrian fatalities of 12.7% is comparable with previous reports of motor vehicle collision fatalities. Because of the presence of increased associated injuries, pedestrians have a significantly higher mortality. Severely injured pedestrians are at a similar risk to motor vehicle occupants for a life-threatening injury of the thoracic aorta. PMID- 9867040 TI - The role of computed tomography in selective management of gunshot wounds to the abdomen and flank. AB - OBJECTIVE: To determine whether computed tomography (CT) is an accurate diagnostic modality for the triage of hemodynamically stable patients with gunshot wounds of the abdomen and flank. METHODS: A chart review of 83 trauma patients for whom abdominal CT was used as initial screening. RESULTS: In 53 patients, CT revealed no evidence of peritoneal penetration, and in 15 patients, there was evidence of either peritoneal penetration or liver injury. There were no false results in these patients. Among 15 patients with questionable peritoneal penetration, cavitary endoscopy was performed in 11 and exploratory laparotomy was performed in 3, and 1 patient was initially observed and subsequently underwent exploratory surgery for a missed colonic injury. CONCLUSION: In selected centers and in hemodynamically stable patients with abdominal and flank gunshot wounds, abdominal CT can be an effective and safe initial screening modality to document the presence or absence of peritoneal penetration and to manage nonoperatively stable patients with liver injuries. If there is any question of peritoneal penetration, cavitary endoscopy should be part of the protocol of nonoperative management. PMID- 9867042 TI - Pulmonary tractotomy as an abbreviated thoracotomy technique. AB - BACKGROUND: Operative abbreviated thoracotomy techniques in thoracic trauma include emergency center thoracotomy, ligation of major arterial branches, packing the thoracic cavity for diffuse bleeding, towel clip or Bogota bag closure of the chest, and pulmonary tractotomy. Pulmonary tractotomy with selective vascular ligation was originally described for deep through-and-through lung injuries that did not involve hilar vessels or airways. Pulmonary tractotomy has evolved into use as an abbreviated thoracotomy technique in patients with severe thoracic or multivisceral trauma. As with any operative technique in high risk patients, specific procedure-related complications may occur and are analyzed herein. The objective of this manuscript is to review the indications, techniques, and results for pulmonary tractotomy in trauma patients requiring abbreviated thoracotomy. METHODS: Medical records were retrospectively reviewed for 30 of 32 consecutive tractotomy patients treated at Ben Taub General Hospital, during a 3-year period. By using a model for logistic regression analysis, the characteristics of each patient and their clinical course were tested for impact on mortality. RESULTS: Seventy percent of patients had at least one intraoperative parameter indicative of acidosis (pH < 7.2), coagulopathy (prothrombin time > 13.8 or partial thromboplastin time > 38.0 seconds), or hypothermia (core temperature < 34 degrees C), and 50% of patients manifested two of these three parameters. The mortality rate among the 30 patients was 17%. Three of the five patients who died were noted to be acidotic, coagulopathic, and hypothermic. Twelve of 25 patients who survived more than 1 day had at least one thoracic complication. There were no late deaths. There was one failed tractotomy and one missed injury. A second thoracotomy was not required for control of a lung injury in any patient. Logistic regression analysis showed that intraoperative blood loss was the only predictive factor for mortality. CONCLUSION: Pulmonary tractotomy is a simple and effective technique in injured patients who require an abbreviated thoracotomy and has an acceptable mortality and complication rate. This follow-up report notes that as definitive therapy, tractotomy continues to allow for direct control of bleeding and air leak and obviates the need for formal resection. PMID- 9867043 TI - Bradycardia and hypotension associated with severe hemorrhage are reversed by morphine given centrally or peripherally in anesthetized rats. AB - BACKGROUND: Severe simple hemorrhage (blood loss in the absence of tissue damage and nociception) leads to a reflex bradycardia and hypotension. Earlier studies showed that this reflex can be attenuated by prior administration of morphine. However, some patients may receive morphine, e.g., for analgesia after they have suffered severe hemorrhage. The aim of this study was to determine whether an established bradycardia and hypotension could be reversed by morphine. METHODS: Four groups of male Wistar rats (236-258 g) were anesthetized with alphadolone/alphaxalone (16-19 mg x hg x h(-1) intravenously). All groups received a hemorrhage of 40% total blood volume (BV) at 2% BV x min(-1). After the loss of 27% BV, bradycardia and hypotension were established equally in groups I and II and III and IV. Groups I (n=8) and III (n=10) received 0.9% saline (20 microL intracerebroventricularly or 1 mL x kg(-1) intravenously, respectively), whereas groups II (n=10) and IV (n=10) received morphine (10 microg intracerebroventricularly or 0.5 mg x kg(-1) intravenously, respectively). RESULTS: In groups I and III, heart rate and mean arterial blood pressure continued to fall, whereas the bradycardia was completely reversed and the hypotension partly reversed in groups II and IV after treatment with morphine. CONCLUSION: Morphine, administered centrally or peripherally, can reverse the bradycardia and markedly can attenuate the hypotension associated with severe hemorrhage. However, any benefit may be more apparent than real because other studies suggest that mortality may be increased. PMID- 9867044 TI - Acute psychosocial impact of pediatric orthopedic trauma with and without accompanying brain injuries. AB - BACKGROUND: The acute psychosocial effects of orthopedic injuries on children and their families are poorly understood. Previous studies have relied on retrospective reports or failed to take into account accompanying brain injuries. The purpose of the present study was to examine prospectively the psychosocial impact of pediatric orthopedic traumatic fractures with and without accompanying brain injuries. METHODS: Participants were 108 children 6 to 12 years old with orthopedic injuries requiring hospitalization: group 1 (n=80) had fractures only, group 2 (n=28) also had moderate or severe brain injuries. Using standardized measures and parent interviews, we obtained preinjury estimates of family functioning and child behavior problems and postinjury measures of parental distress, family stresses, and child behavior. RESULTS: Parents reported significant clinical distress (35% in group 1, 57% in group 2), family burdens (group 2 > group 1), and child behavioral changes (41% in group 1, 89% in group 2). Multiple regression analyses indicated that preinjury family status and brain injuries predicted postinjury parental and family distress. CONCLUSION: Pediatric orthopedic injuries have greater social effects on children with accompanying brain injuries and poorer preinjury family functioning. PMID- 9867045 TI - Closed management and percutaneous fixation of unstable proximal humerus fractures. AB - BACKGROUND: In dealing with displaced proximal humerus fractures, there is still much controversy in treatment modalities. The latest investigations emphasize the concept of minimal exposure and rigid fixation. METHODS: The technique of closed reduction and percutaneous fixation with cannulated screws and k-pins was performed on 19 patients with two- and three-part proximal humerus fractures. The outcomes were evaluated with a mean follow-up of 21 months. RESULTS: All except one case had a solid union of the fracture. Sixteen of 19 patients (84%) acquired good or excellent results according to Neer's classification. No further collapse or avascular necrosis was found. CONCLUSION: The method of closed reduction and percutaneous fixation, although technically demanding, yields satisfactory results in displaced proximal humerus fracture. Cannulated screws provided rigid fixation that was the underlying tenet for early functional retrieval. PMID- 9867046 TI - Treatment of closed humeral shaft fractures with intramedullary elastic nails. AB - PURPOSE: Our purpose is a retrospective study of the results of diaphyseal humeral fracture treatment by elastic intramedullary nailing in comparison with plate osteosynthesis. METHODS: Thirty cases were reviewed, with an average follow up of 2 years. In 14 cases, flexible nails were used, and in the remaining 16 cases, A-O plates were used. We considered patients' sex and age, fracture type according to the A-O classification, type of treatment, consolidation defects, additional therapeutic procedures, healing time, and functional recovery. The surgical technique of elastic nailing appeared very simple, safe, and rather atraumatic because the nail is introduced in a retrograde manner and does not need proximal interlocking. RESULTS: The results of elastic nailing, in terms of fracture healing time and functional recovery, appeared comparable with the results of plating, and complications appeared milder. CONCLUSION: We conclude that intramedullary elastic nailing is valid for routine use in appropriately selected humeral shaft fractures. PMID- 9867047 TI - Locked nailing for displaced surgical neck fractures of the humerus. AB - OBJECTIVE: To report experience with a newly devised humeral locked nail for treating surgical neck fractures of the humerus. This device has the advantages of a small diameter for minimal tissue trauma and transfixing locking screws for reliable fixation. METHODS: From 1993 to 1996, 21 consecutive severely displaced surgical neck fractures of the humerus were antegrade nailed with humeral locked nails; 2 fractures were associated with dislocation and 1 fracture was a comminuted metaphyseal fracture with a failed plating. The average age of patients was 65.8 years; average follow-up time was 19.2 months. The proximal screws were applied upward in 5 patients and downward in 16 patients. Static locking was performed in 8 patients, dynamic locking in 13 others. RESULTS: The average operation time was 55 minutes. No patients needed blood transfusion. All fractures eventually achieved union with an average time to union of 14.8 weeks. On the basis of Neer criteria for outcome analysis, excellent or satisfactory results were obtained for 86% of the patients (18 of 21 patients). No patients had deep infection, implant failure, malunion, osteonecrosis, or nail migration that interfered with joint motion. Due to technique errors, one patient had shoulder joint impingement caused by protrusion of the proximal nail tip. CONCLUSION: The operative method reported here has the advantages of minimal tissue trauma, minimal hardware application, sufficient fixation, and easy operative technique, and it can be a worthy alternative for the treatment of severely displaced surgical neck fractures of the humerus. PMID- 9867048 TI - Out-of-hospital cervical spine clearance: agreement between emergency medical technicians and emergency physicians. AB - OBJECTIVE: Determine the level of agreement between emergency medical technicians (EMTs) and emergency physicians (EPs) when applying an existing emergency medical services/fire department protocol for out-of-hospital clinical cervical spine injury (CSI) clearance in blunt trauma patients. METHODS: Prospective observational study of consecutive blunt trauma patients transported by emergency medical services/fire department during a 3-month study period. The setting was an urban Level I trauma center. Measurement of interrater agreement (kappa) was determined. RESULTS: Mean age of the 190 patients was 34+/-19 years (range, 6 -98 years). Fifty-nine percent of the patients were male. One hundred forty-six patients (77%) were immobilized by EMTs; 17 of these patients were clinically cleared by EPs. Forty-four patients (23%) were clinically cleared by EMTs and presented without CSI precautions; of these, 61% (27 of 44) were immobilized by EPs and 57% (25 of 44) had cervical spine radiographs obtained. Overall, 141 patients (74%) required radiographic clearance. CSI were detected in five patients (2.6%); all five were immobilized in the out-of-hospital setting. Overall disagreement between EMTs and EPs regarding out-of-hospital CSI clearance occurred in 44 patients (23%) (kappa=0.29; 95% confidence interval, 0.15-0.43; p < 0.01). CONCLUSION: Significant disagreement in clinical CSI clearance exists between EMTs and EPs. Further research and education is recommended before widespread implementation of this practice. PMID- 9867049 TI - Measurement of net whole-body transcapillary fluid transport and effective vascular compliance in humans. AB - BACKGROUND: Net whole-body transcapillary fluid transport (TFT) between the circulation and the interstitial (extravascular) space may be calculated as: IV - deltaPV - UV - IL, where IV=infused or ingested volume (when applicable), deltaPV = change in plasma volume, UV=urine volume, and IL=insensible loss. RESULTS: Infusion of 30 mL/kg isotonic saline over 25 minutes increased supine TFT from a basal capillary reabsorption of -106+/-24 mL/h (mean+/-SE) to a net filtration of 1,229+/-124 mL/h. One hour after infusion, reabsorption of -236+/-102 mL/h was seen, and control reabsorption levels returned by 3 hours. Four hours of 30 mm Hg lower body negative pressure (LBNP) elicited no net TFT, probably because of upper body reabsorptive compensation for lower body capillary filtration. When ingestion of 1 L of isotonic saline accompanied LBNP, filtration of 145+/-10 mL/h occurred. Reabsorption of extravascular fluid into the circulation always followed LBNP. CONCLUSION: Application of this technique could aid understanding of physiologic conditions, experimental interventions, disease states, and therapies that cause or are influenced by fluid shifts between intravascular and interstitial compartments. PMID- 9867050 TI - Prosecution and conviction of the injured intoxicated driver. AB - BACKGROUND: Previous studies have reported low conviction rates for drunk drivers injured in motor vehicle crashes and transported to the hospital. The purpose of this study was to evaluate this rate during a recent period and to investigate the variables that predict alcohol-related convictions for injured drunk drivers admitted to our hospital. METHODS: A retrospective review of medical records from January 1991 through May 1997 identified 71 patients who were legally intoxicated drivers injured in motor vehicle crashes. Court records, police reports, and driving records were also obtained. RESULTS: Overall, 51% of the drunk drivers were convicted of alcohol-related offenses and 32% escaped without any conviction. Blood alcohol level and a police officer's estimation of whether the driver had been drinking were significant predictors of an alcohol-related conviction. Age, Injury Severity Score, a police officer's estimation of injury, and the number of people or cars involved in the crash were not significantly associated with legal outcome. CONCLUSION: Although this study shows an important increase in alcohol-related conviction rates, responsibility for further progress will depend on the medical community, law enforcement agencies, and the judicial system working together. PMID- 9867051 TI - The effect of clysed and topical epinephrine on intraoperative catecholamine levels. AB - BACKGROUND: Epinephrine administration for hemostasis during burn wound excision may produce potential anesthetic risks. Two patient groups were studied to determine the absorption of either topical concentrated epinephrine or exogenously injected dilute epinephrine. METHODS: For the topical group (10 patients, 10 procedures), excision of wounds under tourniquet was performed, followed by epinephrine (1 mg/10 mL) gauze with pressure wrapping. For the clysis group (9 patients, 12 procedures), donor sites were injected with 0.5 mg epinephrine/1,000 mL lactated Ringer's solution before harvest. Nine intraoperative serum samples were collected and frozen during each procedure for epinephrine and norepinephrine assay. RESULTS: Concentrated epinephrine (67 mL) was topically applied to excise 1,362 cm2. Dilute epinephrine (1,350 mL) was clysed to obtain 1,950 cm2 autograft. No significant increases in the serum catecholamines or changes in the cardiovascular profiles occurred. CONCLUSION: The administration of either topical or clysed epinephrine during acute burn excision does not cause any side effects for safe anesthetic management; there were no detectable increased plasma levels of epinephrine or norepinephrine. Epinephrine provides the burn surgeon with two safe methods for controlling intraoperative blood loss. PMID- 9867052 TI - Disruptive forces for swine heart, liver, and spleen: their breaking stresses. AB - OBJECTIVES: The mechanical strengths of the swine heart, liver, and spleen were determined by measuring the force that initiated the rupture of the organs. METHODS: The force was loaded by the compression of a detection bar onto the organ, and the breaking stress was calculated by dividing the force by the bar's contact area. The forces were measured with five measurements in five of each type of organ with and without the serous membrane (SM). RESULTS: The breaking stresses of the heart were 14.1+/-0.7 kg/cm2 (mean+/-SEM) with intact epicardium and 8.1+/-0.6 without it; those of the spleen were 9.0+/-0.9 with its capsule and 1.8+/-0.1 without the capsule, and those of the liver were 4.6+/-0.3 with and 3.6+/-0.3 without the SM. A three-way layout analysis of variance showed that the organs, their SM and individual swine were independent determinants of the breaking stresses. CONCLUSION: With the intact SM, the heart was strongest and the liver was weakest. When the SM was not present, the spleen lost 80% of its strength, the liver 22% of its strength, and the heart lost 43% of its strength. PMID- 9867053 TI - Population-based analysis of 10,766 hospitalizations for mandibular fractures in California, 1991 to 1993. AB - BACKGROUND: This study describes mandibular fracture incidence, causes, and consequences in a large population. METHODS: California hospital discharge data from 1991 to 1993 were examined to describe causes, lengths of stay, and hospital charges for patients hospitalized for mandibular fractures. Rates were calculated per 100,000 population. RESULTS: There were 10,766 discharges with mandibular fracture as principal diagnosis or as diagnoses two through five on the discharge record (rate=11.5). Of these, 2,694 had mandibular fracture as the only diagnosis (principal diagnosis). For all 10,766 cases, assaults were responsible for more than half (54.0%) of all admissions for mandibular fracture. The highest rates were found among males (18.7), blacks (43.0), and adults aged 16 to 20 (26.5). Charges for the initial hospitalization (excluding physician's fees) for the 2,694 cases with only a diagnosis of mandibular fracture were used to estimate mean charges ($8,740). The total extrapolated 1993 inflation-adjusted hospital charges for mandibular fractures were $34 million per year. Most patients' bills were submitted to government payers, such as Medicaid. CONCLUSION: Treatment of mandibular fractures represents a considerable cost to public-supported programs as well as to patients. PMID- 9867054 TI - Steroids and spinal cord injury: revisiting the NASCIS 2 and NASCIS 3 trials. AB - The National Acute Spinal Cord Injury Study (NASCIS) 2 and 3 trials are often cited as evidence that high-dose methylprednisolone is an efficacious intervention in the management of acute spinal cord injury. Neither of these studies convincingly demonstrate the benefit of steroids. There are concerns about the statistical analysis, randomization, and clinical end points. Even if the putative gains are statistically valid, the clinical benefits are questionable. Furthermore, the benefits of this intervention may not warrant the possible risks. This paper comments on these two clinical trials. PMID- 9867055 TI - Traumatic choledochogastric fistula: endoscopic evaluation and treatment with a biliary stent. PMID- 9867056 TI - Use of titanium vascular staples in trauma. PMID- 9867057 TI - Primary fusion of the talonavicular joint after fracture dislocation of the navicular bone. PMID- 9867058 TI - Fentanyl should be used with caution in patients with severe brain injury. PMID- 9867059 TI - Balloon tamponade for bleeding control in penetrating liver injuries. PMID- 9867061 TI - Progress in diabetes care: introduction. PMID- 9867062 TI - Pathophysiology of diabetic nephropathy. AB - Diabetic nephropathy (DN) is now the commonest cause of end-stage renal failure in the Western world. Recent studies examining the pathogenesis of diabetic complications have focused on the complex interaction between genetic and hemodynamic mechanisms in addition to metabolic factors such as advanced glycation, protein kinase C (PKC) activation, and polyol production. The importance of the various components, particularly with regard to the progression of DN, is currently being explored with the assistance of targeted drug intervention studies. PMID- 9867063 TI - Role of angiotensin-converting enzyme inhibition in glucose metabolism and renal injury in diabetes. AB - The role of angiotensin-converting enzyme (ACE) inhibition in glucose metabolism and renal injury in diabetes has been extensively investigated in diabetic humans, as well as in animal models of diabetes. Accumulated data indicate that ACE inhibitors have either no adverse effect on glucose control or insulin sensitivity or may even improve them. ACE inhibitors also appear to have neutral or positive effects on lipid metabolism. The variability of results between studies may relate to differences in experimental design, the degree of glycemia or insulin resistance, potassium balance, and dose or duration of ACE inhibitor treatment, among others. In contrast, ACE inhibitors have proved effective in limiting proteinuria and retarding renal function loss in insulin-dependent diabetes mellitus (IDDM) or non-insulin-dependent diabetes mellitus (NIDDM) patients. In rats with experimental or spontaneous diabetes, ACE inhibitors also reduce proteinuria and limit glomerular as well as tubulointerstitial damage, independent of their effects on systemic arterial pressure. How ACE inhibitors limit renal injury in diabetes is not entirely clear, but hemodynamic and nonhemodynamic mechanisms may be involved. Increasing evidence suggests that the intrarenal renin-angiotensin system (RAS) may be altered or activated in the diabetic kidney. Such activation may be specifically inhibited by ACE inhibitors and may explain the superiority of this class of agents over other antihypertensive agents in reducing proteinuria and slowing the progression of diabetic nephropathy. PMID- 9867064 TI - Effects of perindopril on renal histomorphometry in diabetic subjects with microalbuminuria: a 3-year placebo-controlled biopsy study. AB - We conducted a 3-year randomized placebo-controlled double-blind study to determine the effects of the angiotensin-converting enzyme (ACE) inhibitor perindopril (PE) on the progress of renal function and histology in subjects with diabetes and microalbuminuria. Forty non-insulin-dependent (NIDDM) and insulin dependent (IDDM) diabetic subjects, either normotensive or hypertensive, were randomly assigned to receive PE (n = 20) or placebo (n = 20). A percutaneous renal biopsy was performed initially in all patients and repeated in 29 patients after 3 years. The mean glomerular volume, glomerular basement membrane (GBM) thickness, interstitial fibrosis, sclerosed glomeruli, and volume fraction of capillary lumina were measured histomorphometrically. Before treatment, both groups had similar clinical characteristics, blood pressure, glycosylated hemoglobin (Hb), albumin excretion rate, glomerular filtration rate (GFR), serum creatinine, and renal structural damage. Blood pressure was well controlled in both groups. After 3 years' therapy, there was no significant change in renal function and albuminuria in the PE or placebo groups. The increase in GBM thickness in nine paired biopsies was significantly less in PE-treated subjects (P = .0275). Interstitial fibrosis tended to increase less in the PE group, although this did not reach statistical significance. This study indicates that long-term therapy with PE may decrease or delay the progression of structural glomerular damage in microalbuminuric diabetic subjects. PMID- 9867065 TI - Diabetes mellitus and vascular lesions. AB - Cardiovascular complications represent by far the most severe manifestations of diabetes mellitus. Treatment aimed at stopping progression of vascular lesions may fall short if initiated when the disease becomes clinically evident. Therefore, identification of the earliest vascular disfunctions may offer the best opportunity to interfere with pathogenic mechanisms and avoid progression of diabetic vasculopathy. In this report, we present a few mechanisms that alter hemodynamic and metabolic homeostasis in the course of diabetes mellitus. Endothelial function with special emphasis on nitric oxide and oxidative stress, advanced glycation end products, and the renin angiotensin system are briefly discussed. New pharmacological agents that may favorably influence these parameters are presently undergoing clinical trials. However, tight control of plasma glucose and cardiovascular risk factors represent the cornerstone of the treatment in diabetes to slow progression of vascular disease. PMID- 9867066 TI - Effects of angiotensin-converting enzyme inhibition on vascular remodeling of resistance vessels in hypertensive patients. AB - Essential hypertension is known to be associated with a decrease in the lumen diameter and an increase in the wall thickness to lumen diameter ratio of the resistance vessels. Recently, it has been clarified that this alteration does not necessarily involve vascular growth, but could be due to a rearrangement of the same amount of material, a phenomenon now termed "eutrophic remodeling." These changes are found both in human essential hypertension and in animal models of genetic hypertension. Antihypertensive treatment with angiotensin-converting enzyme (ACE) inhibitors causes a dose-dependent regression of the media to lumen ratio in rats. Clinical studies have now confirmed these findings, showing that when previously untreated essential hypertensive patients are treated with the ACE inhibitor perindopril (PE), the abnormal structure of resistance vessels regresses toward normal values; in contrast, treatment with a beta-blocker does not affect the abnormal vascular structure. The available evidence thus indicates that ACE inhibitors are able to normalize the abnormal resistance vessel structure in essential hypertension, and suggest that this effect may not only be dependent on their ability to reduce blood pressure. This review summarizes these findings, and discusses the extent to which this is desirable. PMID- 9867067 TI - Attenuation of diabetes-associated mesenteric vascular hypertrophy with perindopril: morphological and molecular biological studies. AB - Vascular disease is now the major cause of morbidity and mortality in the diabetic population. Our group explored the vascular changes associated with experimental diabetes and examined whether these changes can be ameliorated by angiotensin-converting enzyme (ACE) inhibition. The ACE inhibitor perindopril (PE) was administered to streptozotocin-induced diabetic rats for 24 weeks. At death, mesenteric vessels were perfused in vivo followed by assessment of the vascular architecture by quantitative histomorphometry. In a subgroup of animals, RNA was extracted from the mesenteric vasculature for assessment of gene expression of the prosclerotic cytokine, transforming growth factor beta 1 (TGFbeta1), and the matrix protein, type IV collagen. Diabetes was associated with smooth muscle hypertrophy and extracellular matrix (ECM) accumulation. ECM accumulation, particularly collagen deposition, was observed in the medial and adventitial layers. ACE inhibition prevented mesenteric vascular hypertrophy after 24 weeks of diabetes. In addition, overexpression of TGFbeta1 in the vessels of diabetic animals was prevented by PE treatment. Similarly, type IV collagen mRNA levels were increased in diabetic vessels, and this overexpression was also prevented by PE therapy. In summary, ACE inhibition attenuates many of the vascular changes observed in experimental diabetes and may have important clinical implications as a vasoprotective agent in human diabetes. PMID- 9867068 TI - Effect of angiotensin-converting enzyme inhibition with perindopril and beta blockade with atenolol on retinal blood flow in hypertensive diabetic subjects. AB - The effect of angiotensin-converting enzyme (ACE) inhibitors on the diabetic retinal circulation has not been studied previously. The aim of this study was to evaluate the effect of ACE inhibition and beta-blockade on retinal blood flow (RBF) in a group of 45 hypertensive diabetic subjects using a randomized double blind trial over a period of 12 months. Laser Doppler velocimetry and computed image analysis were used to measure RBF. The changes in blood pressure over 12 months were comparable (perindopril [PE]: systolic [SBP] 152.1 +/- 3.3 and diastolic [DBP] 97.2 +/- 1.7 mm Hg to SBP 136.8 +/- 3.4 and DBP 85.8 +/- 2.1; atenolol: SBP 158.9 +/- 5.1 and DBP 97.5 +/- 1.6 mm Hg to SBP 137.9 +/- 3.4 and DBP 85.1 +/- 1.6; P = .607, mean +/- SEM). RBF decreased from 17.19 +/- 2.21 microL x min(-1) to 14.18 +/- 1.50 microL x min(-1) in the PE group (n = 15, P = .208) while it increased with atenolol from 15.80 +/- 1.24 microL x min(-1) to 16.99 +/- 1.18 microL x min(-1) (n = 17, P = .399). The comparison of percentage changes in RBF (PE -7.16% +/- 11.49%; atenolol, +15.31% +/- 9.51%) reached statistical significance (P < .05). There was an increase in RBF in 33.3% of subjects receiving PE and in 70.6% of those receiving atenolol. Similar trends were found for retinal conductance. There were no significant changes in the parameters of retinal vascular permeability. Albuminuria decreased to a greater degree with PE, but did not reach significance (PE, 112.1 +/- 39.5 mg/24 h to 88.6 +/- 30.5 mg/24 h; atenolol, 87.3 +/- 51.7 mg/24 h to 82.1 +/- 47.7 mg/24 h). This suggests that ACE inhibition therapy may promote a hemodynamic milieu in the hypertensive diabetic retinal circulation that serves to protect against the progression of diabetic retinopathy, whereas beta-blockade has the opposite effect. PMID- 9867069 TI - Whole-body protein metabolism assessed by leucine and glutamine kinetics in adult patients with active celiac disease. AB - To assess the effect of increased renewal of intestinal epithelial cells on leucine and glutamine (Gln) turnover, 4-hour intravenous infusions of L-[1 (13)C]leucine and L-[2-(15)N]Gln were administered to five adult patients with active celiac disease in the postabsorptive state. There was a 35% increase in leucine flux (micromoles per kilogram per hour) in patients (117 +/- 17) compared with healthy controls (96 +/- 11, P < .03). Gln flux was increased by 13% in patients (377 +/- 35) versus controls (335 +/- 16, P < .04). These results suggest that active celiac disease, characterized by villous atrophy and crypt cell hyperplasia, is associated with a dramatic increase in whole-body protein breakdown as assessed by 13C-leucine, which may contribute per se to the protein malnutrition status of the patients. The increase in Gln utilization as assessed by L-[2-(15)N]Gln was moderate, but may have been offset due to the villose atrophy and ensuing reduced intestinal epithelial cell mass. The results are consistent with the concept that increased renewal of intestinal epithelial cells represents a sizable fraction of whole-body protein turnover and that Gln is an important fuel for epithelial intestinal cells in vivo. PMID- 9867070 TI - Effects of reduced energy intake on protein utilization in obese children. AB - Dietary treatment of pediatric obesity is a challenge given the need for adequate nutrients to support the maintenance of lean tissue and growth. The primary purpose of this investigation was to assess the effects of reduced energy intake on protein turnover in obese children aged 8 to 10 years. Following a 2-week baseline period, 16 subjects reduced energy intake during a 6-week intervention period. At baseline and following the intervention, 15N-glycine methodology was used to measure nitrogen flux (Q), protein synthesis (PS), protein breakdown (PB), and net turnover ([NET] PS - PB). Other criterion measures included resting metabolic rate (RMR), fat mass (FM), fat-free mass (FFM), urinary creatinine to height ratio (Cr:Ht), and nitrogen balance (NB). On average, subjects lost 2.2 +/ 0.3 kg, of which greater than 85% was FM. Decreased Q (P = .03) indicated downregulation of protein turnover in response to diet-induced weight loss. While PB did not change, NET declined slightly (P = .06) as a consequence of reduced PS (P = .03). Reductions in FFM (P = .09), Cr:Ht (P = .02), and NB (P = .03) accompanied alterations in protein turnover, but there was no change in the RMR. In conclusion, while short-term therapy promoted the loss of FM and did not compromise RMR, practitioners must be cautious when prescribing diets, given the observed changes in protein utilization and somatic protein status. Longitudinal studies are needed to further characterize the metabolic responses of obese children to long-term diet therapy. PMID- 9867071 TI - Relationships between the pituitary-adrenal hormones, insulin, and glucose in middle-aged men: moderating influence of psychosocial stress. AB - We examined whether the relationships between the pituitary-adrenal hormones (corticotropin [ACTH) and cortisol), insulin, and glucose differ as a function of psychosocial stress defined in terms of vital exhaustion (VE) and depressive behavior (DB). The participants were 69 normotensive and 21 unmedicated borderline hypertensive (BH) middle-aged men whose work is stressful. Hormonal and metabolic variables were measured during an oral glucose tolerance test (OGTT), and the cortisol response to dexamethasone (DXM) suppression and intravenous ACTH stimulation was also measured. We found that the basal ACTH level during the OGTT was positively associated with the cortisol response to ACTH at 60 minutes, the fasting insulin level, and the insulin to glucose ratio among exhausted and high DB men, while the reverse was true for nonexhausted and low DB men. Also, a high cortisol response to ACTH, a low cortisol level during the OGTT, and a high ratio of these cortisol determinations (cortisol ratio) were associated with high fasting insulin and glucose levels, the summed insulin values, and the insulin to glucose ratio only among nonexhausted and low DB men; among exhausted and high DB men, these associations were less pronounced, absent, or in the opposite direction. The findings suggest that VE and DB have a moderating influence on the relationships among the hormonal and metabolic parameters studied. Psychosocial stress may affect the pituitary-adrenocortical system in complex ways, contributing thereby to insulin resistance, hyperinsulinemia, and coronary heart disease (CHD) risk. PMID- 9867072 TI - Ketosis resistance in the male offspring of protein-malnourished rat dams. AB - Plasma beta-hydroxybutyrate concentrations were measured in the offspring of rats that were fed either a control (20% protein) diet or low-protein (8% protein) diet during pregnancy and lactation. Low-protein offspring had significantly lower plasma beta-hydroxybutyrate compared with controls in the fed state (P < .04) and after fasting for 24 hours (P < .001) and 48 hours (P < .04). There were no differences in blood glucose, acetoacetate, plasma glucagon, cholesterol, or glycerol between control and low-protein offspring. However, plasma nonesterified fatty acids (NEFAs) were significantly higher in low-protein offspring in the fed state (P < .05). In contrast, plasma triglycerides and insulin were significantly lower in low-protein offspring compared with controls when fed (P < .001) and after a 24-hour fast (P < .001). These results suggest that poor maternal and early postnatal nutrition can have long-term effects on ketone body metabolism in the offspring during adulthood. This apparent ketosis resistance is similar to that observed in some forms of human diabetes. PMID- 9867073 TI - Hormonal regulation of 18S RNA, leptin mRNA, and leptin release in adipocytes from hypothyroid rats. AB - The present studies were designed to examine the regulation of leptin release in primary cultures of adipocytes from fed hypothyroid rats incubated with hormones for 24 hours. Leptin release was increased in the presence of dexamethasone, while the decrease in leptin mRNA content over a 24-hour incubation was reduced by dexamethasone. Dexamethasone did not affect the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA or 18S RNA content of adipocytes. Insulin increased leptin release by adipocytes in both the absence and presence of dexamethasone. Although insulin also prevented the loss of leptin mRNA, this effect was less than that observed for GAPDH mRNA or 18S RNA content. In isolated adipocytes, the loss of almost half the 18S RNA content over a 24-hour incubation was prevented in the presence of insulin but not oxytocin or epidermal growth factor (EGF). The specific beta3 catecholamine agonist CI 316,243 inhibited the effects of dexamethasone on leptin release and leptin mRNA accumulation, as did EGF, without affecting 18S RNA content. Oxytocin inhibited the increase in leptin release due to dexamethasone without affecting leptin mRNA levels. These data indicate that although dexamethasone and insulin are positive regulators of leptin release, only dexamethasone specifically prevented the loss of leptin mRNA in cultured rat adipocytes. In contrast, insulin, but not dexamethasone, prevented the marked loss in 18S RNA observed over a 24-hour incubation of rat adipocytes. PMID- 9867074 TI - Inhibition of lipolysis reduces beta1-adrenoceptor-mediated thermogenesis in man. AB - The purpose of the study was to investigate whether the increase in energy expenditure and lipid oxidation during beta1-adrenergic stimulation is caused by the concomitant increase in lipolysis. Twelve healthy male subjects participated in three trials: no-LIP/-, inhibition of lipolysis by pretreatment with acipimox followed by saline infusion; -/BETA, no pretreatment, with dobutamine infusion to stimulate beta1-adrenoceptors; and no-LIP/BETA, pretreatment with acipimox followed by dobutamine infusion. Inhibition of lipolysis did not affect baseline energy expenditure, but decreased lipid oxidation and increased carbohydrate oxidation. Energy expenditure and lipid oxidation increased significantly during beta1-adrenergic stimulation, but this increase was significantly smaller when lipolysis was inhibited ([baseline v infusion period] energy expenditure: -/BETA, 5.15 +/- 0.16 v 6.11 +/- 0.26 kJ/min, P < .001; no-LIP/BETA, 5.28 +/- 0.17 v 5.71 +/- 0.19 kJ/min, P < .01; lipid oxidation: -/BETA, 0.059 +/- 0.004 v 0.073 +/- 0.006 g/min, P < .01; no-LIP/BETA, 0.034 +/- 0.005 v 0.039 +/- 0.006 g/min, P < .05). Baseline plasma glycerol and nonesterified fatty acid (NEFA) concentrations decreased after inhibition of lipolysis. Glycerol and NEFA increased significantly during beta1-adrenergic stimulation alone (glycerol, 65.0 +/- 5.3 v 117.0 +/- 10.9 micromol/L; NEFA, 362 +/- 24 v 954 /- 89 micromol/L; both P < .001). Concomitant administration of acipimox prevented a substantial part of the increase in lipolysis during beta1-adrenergic stimulation, but the increase in plasma glycerol and NEFA remained significant (glycerol, 40.4 +/- 2.2 v 44.8 +/- 2.2 micromol/L; NEFA, 118 +/- 18 v 160 +/- 19 micromol/L; both P < .05). In conclusion, a reduced availability of plasma NEFA was associated with a reduced increase in energy expenditure and lipid oxidation during beta1-adrenergic stimulation in man. PMID- 9867075 TI - Evidence that endothelin-1 (ET-1) inhibits insulin-stimulated glucose uptake in rat adipocytes mainly through ETA receptors. AB - The specificity of endothelin (ET) receptors involved in the inhibition of insulin-stimulated glucose uptake (ISGU) in rat adipocytes was investigated. Adipocytes were isolated from the epididymal fat pads of Sprague-Dawley rats. To determine receptor subtypes, we used three ET isopeptides, ET-1 and ET-2, both of which are nonselective agonists, and ET-3, a selective agonist for ETC receptors, to displace [125I]ET-1 binding from the fat cells. The efficiency of displacement was ET-1 > ET-2 >> ET-3, indicating that the primary receptors involved belonged to the ETA subtype. At an equal concentration of 1 micromol/L, BQ-610, a selective ETA antagonist, displaced [125I]ET-1 from binding to fat cells, whereas IRL-1038, a selective ETB antagonist, did not. Using [3H]2-deoxy-D-1-glucose ([3H]2-DG) as a tracer in studies of glucose uptake, we found that equimolar BQ 610 completely reversed the inhibitory effect of ET-1 on ISGU, whereas IRL-1038 was ineffective. Northern blot analysis of adipocyte receptors showed abundant mRNA for ETA, but no ETB subtype. These results clearly demonstrate that ETA is the predominant receptor in rat adipocytes. PMID- 9867076 TI - Muscle damage induced by experimental hypoglycemia. AB - To determine organ damage due to hypoglycemia, we studied the effects of insulin dose and hypoglycemia duration on serum enzyme activity in rabbits. Thirty rabbits were randomly divided into five groups according to hypoglycemia duration and insulin dose: A2, hypoglycemia for 30 minutes with 2 U/kg insulin; A10, hypoglycemia for 30 minutes with 10 U/kg insulin; B2, hypoglycemia for 60 minutes with 2 U/kg insulin; B10, hypoglycemia for 60 minutes with 10 U/kg insulin; and C, no hypoglycemia with 10 U/kg insulin and 50% glucose. Insulin-induced hypoglycemia was reversed by intravenous injection of glucose. Alterations in serum enzyme activity and creatine kinase (CK) isoenzyme distribution were determined before and after insulin injection. Serum CK activity increased significantly in all hypoglycemic groups compared with preinjection values, and tended to remain high for 24 hours in both groups A10 and B10. Serum activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) increased only in group B10. In addition, the level of band 4 of serum CK isoenzymes, which exists predominantly in skeletal muscle and myocardium, increased significantly in group B10. These results suggest that the increase in both serum enzyme and CK band 4 isoenzyme activities during hypoglycemia is primarily due to damage in muscle rather than liver, and that the hypoglycemia duration and insulin dosage may influence the extent of organ damage. PMID- 9867077 TI - Potential benefit of inhibitors of advanced glycation end products in the progression of type II diabetes: a study with aminoguanidine in C57/BLKsJ diabetic mice. AB - Prolonged hyperglycemia in type II diabetic patients is linked both with diabetic complications and with further impairment of glucose homeostasis, possibly due to glucose toxicity of the beta cell. While the connection between the accumulation of extracellular advanced glycation end products (AGEs) and the development of complications is well established, it has only recently been suggested that intracellular glycation may be equally adverse and could be involved in the pathogenesis of glucose toxicity in vitro. Aminoguanidine is a recognized inhibitor of the formation of both extracellular and intracellular AGEs. In this study, we show that the development of diabetes, measured by increased water intake and concomitant midday blood glucose levels in type II genetically diabetic mice, is reduced by treatment with aminoguanidine at a dosage of 500 mg/kg/d for 12 weeks in the diet. In addition, at the end of the study, aminoguanidine reduced the decline in serum and pancreatic insulin levels and the degree of pancreatic islet morphological degeneration, all of which are associated with pancreatic insufficiency following prolonged hyperglycemia in this animal model. These results suggest that AGEs may be involved in the aggravation of type II diabetes in vivo and aminoguanidine may be beneficial in its treatment. PMID- 9867078 TI - Effects of low-dose recombinant human insulin-like growth factor-I on insulin sensitivity, growth hormone and glucagon levels in young adults with insulin dependent diabetes mellitus. AB - Despite recent interest in the therapeutic potential of recombinant human insulin like growth factor-I (rhIGF-I) in the treatment of diabetes mellitus, its mechanism of action is still not defined. We have studied the effects of low-dose bolus subcutaneous rhIGF-I (40 microg/kg and 20 microg/kg) on insulin sensitivity, growth hormone (GH) and glucagon levels in seven young adults with insulin-dependent diabetes mellitus (IDDM) using a randomized double-blind placebo-controlled crossover study design. Each was subjected to a euglycemic clamp (5 mmol/L) protocol consisting of a variable-rate insulin infusion clamp (6:00 PM to 8:00 AM) followed by a two-dose hyperinsulinemic clamp (insulin infusion of 0.75 mU x kg(-1) x min(-1) from 8 to 10 AM and 1.5 mU x kg(-1) x min( 1) from 10 AM to 12 noon) incorporating [6,6 2H2]glucose tracer for determination of glucose production/utilization rates. Following rhIGF-I administration, the serum IGF-I level (mean +/- SEM) increased (40 microg/kg, 655 +/- 90 ng/mL, P < .001; 20 microg/kg, 472 +/- 67 ng/mL, P < .001; placebo, 258 +/- 51 ng/mL). Dose related reductions in insulin were observed during the period of steady-state euglycemia (1 AM to 8 AM) (40 microg/kg, 48 +/- 5 pmol/L, P = .01; 20 microg/kg, 58 +/- 8 pmol/L, P = .03; placebo, 72 +/- 8 pmol/L). The mean overnight GH level (40 microg/kg, 9.1 +/- 1.4 mU/L, P = .04; 20 microg/kg, 9.6 +/- 2.0 mU/L, P = .12; placebo, 11.3 +/- 1.7 mU/L) and GH pulse amplitude (40 microg/kg, 18.8 +/- 2.9 mU/L, P = .04; 20 microg/kg, 17.0 +/- 3.4 mU/L, P > .05; placebo, 23.0 +/- 3.7 mU/L) were also reduced. No differences in glucagon, IGF binding protein-1 (IGFBP-1), acetoacetate, or beta-hydroxybutyrate levels were found. During the hyperinsulinemic clamp conditions, no differences in glucose utilization were noted, whereas hepatic glucose production was reduced by rhIGF-I 40 microg/kg (P = .05). Our data demonstrate that in subjects with IDDM, low-dose subcutaneous rhIGF-I leads to a dose-dependent reduction in the insulin level for euglycemia overnight that parallels the decrease in overnight GH levels, but glucagon and IGFBP-1 levels remain unchanged. The decreases in hepatic glucose production during the hyperinsulinemic clamp study observed the following day are likely related to GH suppression, although a direct effect by rhIGF-I cannot be entirely discounted. PMID- 9867079 TI - Growth hormone bioactivity, insulin-like growth factors (IGFs), and IGF binding proteins in obese children. AB - In obese children, both spontaneous and stimulated growth hormone (GH) secretion are impaired but a normal or increased height velocity is usually observed. This study was undertaken to explain the discrepancy between impaired GH secretion and normal height velocity. We evaluated the GH bioactivity (GH-BIO), GH serum level by immunofluorimetric assay (GH-IFMA), insulin-like growth factor-I (IGF-I), IGF II, and IGF binding protein-1 (IGFBP-1), IGFBP-2, and IGFBP-3 in 21 prepubertal obese children (13 boys and eight girls) aged 5.7 to 9.4 years affected by simple obesity and in 32 (22 boys and 10 girls) age- and sex-matched normal-weight controls. The results were as follows (obese versus [v] controls): GH-IFMA, 4.84 +/- 3.54 v 23.7 +/- 2.04 microg/L (P < .001); GH-BIO, 0.60 +/- 0.45 v 1.84 +/- 0.15 U/mL (P < .001); IGF-I, 173.8 +/- 57.2 v 188.6 +/- 132.6 ng/mL (nonsignificant); IGF-II, 596.1 +/- 139.7 v 439.3 +/- 127.4 ng/mL (P < .001); IGFBP-1, 23.25 +/- 14.25 v 107 +/- 165.7 ng/mL (P < .05); IGFBP-2, 44.37 +/- 62.18 v 385.93 +/- 227.81 ng/mL (P < .001); IGFBP-3, 3.31 +/- 0.82 v 2.6 +/- 0.94 microg/mL (P < .05); and IGFs/IGFBPs, 1.32 +/- 0.32 v 1.07 +/- 0.34 (P < .05). In conclusion, in prepubertal obese children, not only immunoreactive but also bioactive GH concentrations were low. In these subjects, therefore, nutritional factors and insulin may contribute to sustain normal growth also by modulating several components of the IGF-IGFBP system. PMID- 9867080 TI - Subcutaneous or visceral adipose tissue expression of the PPARgamma gene is not altered in the fatty (fa/fa) Zucker rat. AB - We cloned 537 basepairs (bp) of rat partial peroxisome proliferator-activated receptor gamma2 (PPARgamma2) cDNA and examined the effect of fasting or obesity on the expression of two isoforms of rat PPARgamma, gamma1 and gamma2, in either subcutaneous or mesenteric adipose tissue specimens using an RNase A protection assay. In Wistar rats, expression of both isoforms was dramatically reduced after 48 hours of fasting in the two fat tissue specimens. In comparing genetically obese (fa/fa) Zucker rats and lean control rats, no significant difference was observed in expression of the two isoforms in either type of adipose tissue. From these findings, we conclude that the adipose tissue level of rat PPARgamma depends on nutritional deprivation but is not closely associated with either obesity or insulin resistance in obese Zucker rats. PMID- 9867081 TI - Dietary specific sugars for serum protein enzymatic glycosylation in man. AB - All glycoprotein sugars can theoretically derive from glucose. However, dietary specific sugars could represent preferential substrates or have regulatory roles in enzymatic glycosylation. This hypothesis was tested in man using stable isotopes. Healthy subjects ingested different amounts (150, 300, or 550 mg) of artificially 13C-enriched sugar (galactose, mannose, or glucose) diluted in 200 mL water containing 50 g 13C-poor sucrose. 13C enrichment of expired CO2 was monitored for 8 hours during indirect calorimetry. Serum glycoproteins were precipitated and delipidated at various intervals. Glycoprotein neutral sugars were obtained by acidic hydrolysis, purified by ion-exchange chromatography, derivatized to alditol acetates, and analyzed by gas chromatography-isotope ratio mass spectrometry. The oxidation rate for galactose and mannose was slower than the rate for glucose. Total oxidation over the 8-hour period was less than 10% of the ingested amount of galactose or mannose. Galactose and mannose were readily incorporated into glycoprotein glycans, in the native form or after interconversion, despite ingestion of a large excess of sucrose: glycoprotein sugar 13C enrichment was strongly higher after 13C-galactose or 13C-mannose than after 13C-glucose. Thus, the metabolism of these three sugars appears to be different. Specific dietary sugars could represent a new class of non essential nutrients displaying interesting metabolic roles. This could have practical consequences especially in parenteral nutrition, where glucose is currently the only sugar available for metabolism. PMID- 9867082 TI - Effect of tamoxifen on cholesterol synthesis in HepG2 cells and cultured rat hepatocytes. AB - The objective of this study was to investigate the mechanisms by which tamoxifen modifies cholesterol metabolism in cellular models of liver metabolism, HepG2 cells and rat hepatocytes. The effect of tamoxifen on cholesterol and triglyceride-palmitate synthesis was measured using isotopomer spectral analysis (ISA) and gas chromatography-mass spectrometry (GC-MS) and compared with the effects of progesterone, estradiol, the antiestrogen ICI 182,780, and an oxysterol, 25-hydroxycholesterol (25OHC). Cholesterol synthesis in cells incubated in the presence of either [1-(13)C]acetate, [U-13C]glucose, or [4,5 (13)C]mevalonate for 48 hours was reduced in the presence of 10 micromol/L tamoxifen and 12.4 micromol/L 25OHC in both HepG2 cells and rat hepatocytes. The ISA methodology allowed a clear distinction between effects on synthesis and effects on precursor enrichment, and indicated that these compounds did not affect enrichment of the precursors of squalene. Progesterone was effective in both cell types at 30 micromol/L and only in HepG2 cells at 10 micromol/L. Estradiol and ICI 182,780 at 10 micromol/L did not inhibit cholesterol synthesis. None of the compounds altered the synthesis of triglyceride-palmitate in either cell type. Treatment of cells with tamoxifen produced accumulation of three sterol precursors of cholesterol, zymosterol, desmosterol, and delta8 cholesterol. This pattern of precursors indicates inhibition of delta24,25 reduction in addition to the previously described inhibition of delta8 isomerase. We conclude that tamoxifen is an effective inhibitor of the conversion of lanosterol to cholesterol in cellular models at concentrations comparable to those present in the plasma of tamoxifen-treated individuals. Our findings indicate that this mechanism may contribute to the effect of tamoxifen in reducing plasma cholesterol in humans. PMID- 9867083 TI - Serum leptin levels and leptin expression in growth hormone (GH)-deficient and healthy adults: influence of GH treatment, gender, and fasting. AB - Growth hormone (GH) treatment is associated with a reduction in fat mass in healthy and GH-deficient (GHD) subjects. This is mainly mediated via a direct GH action on adipose cells and stimulation of lipolysis. Leptin is secreted from adipose tissue and may be involved in signaling information about adipose tissue stores to the brain. Hormonal regulation of leptin is still not fully elucidated, and in the present study, we investigated both the long-term (4-month) and short term (28-hour) GH effects on serum leptin and leptin gene expression in subcutaneous adipose tissue. In GHD adults (n = 24), leptin correlated with most estimates of adiposity (r = .62 to .86), as previously found in healthy subjects. However, no correlation was observed with intraabdominal fat determined by computed tomographic (CT) scan (INTRA-CT). GH treatment for 4 months had no independent effect on either serum leptin or leptin gene expression. In a short term study, we found that fasting gradually reduced leptin levels in both healthy men and GHD adults, with a maximum reduction of 58% to 60% (P < .01) after 31 hours. No independent effect of GH suppression or GH substitution on serum leptin was found during fasting. Adipose tissue leptin mRNA correlated with serum leptin (r = .51, P < .01) and the body mass index ([BMI] r = .55, P < .05). Serum leptin levels and gene expression were significantly higher in women compared with men (26.6 +/- 5.8 v 10.0 +/- 1.30 ng/mL, P < .05). However, in regression analysis accounting for the gender differences in subcutaneous femoral adipose tissue (FEM CT), the difference in serum leptin disappeared, indicating that subcutaneous femoral fat or factors closely related to femoral fat (eg, sex hormones) may be causal factors for the gender difference in leptin. PMID- 9867084 TI - Effect of a controlled high-fat versus low-fat diet on insulin sensitivity and leptin levels in African-American and Caucasian women. AB - African-American women have been shown to be more insulin-resistant than age- and weight-matched Caucasian women, but the reasons for this difference are unclear. The purpose of the present study was to determine whether experimental manipulation of dietary fat intake has differential effects by race on insulin sensitivity (S(I)) in 20 African-American and 11 Caucasian women. Additionally, leptin levels before and after 3 weeks of an isocaloric high-fat ([HF] 50% fat, 35% carbohydrate, and 15% protein) or low-fat ([LF] 20% fat, 55% carbohydrate, and 15% protein) diet were compared. African-American and Caucasian women did not differ significantly in the body mass index (BMI) or percentage body fat at baseline. S(I) (adjusted for BMI) decreased on the HF diet and increased on the LF diet in both races combined relative to the baseline control (control, 2.42 +/ 0.22; HF, 2.29 +/- 0.22; LF, 2.75 +/- 0.21 x 10(-4) min(-1)/microU x mL; main effect of diet, P = .04). There was a 6% decrease in S(I) on the HF diet compared with the control in women of both races, while the LF diet increased S(I) by 6% in African-American and 20% in Caucasian women. Leptin levels increased by 14% on the HF versus control diet in African-Americans (35.2 +/- 3.0 v 30.8 +/- 3.0 ng/mL, P < .01), but did not change with diet in Caucasian women. Glucose and insulin administration had no effect on leptin levels. We conclude that a HF diet consumed over several weeks reduces S(I) in healthy women of both races; however, the magnitude of increase in S(I) on a LF diet is greater in Caucasian women. The HF diet significantly increased leptin levels in African-American women, although there were no other influences of diet, insulin, or race on serum leptin. PMID- 9867085 TI - No effect of Trp64Arg beta3-adrenoceptor polymorphism on the plasma leptin concentration in Pima Indians. AB - In rodents, administration of leptin promotes beta3-adrenergic stimulation of thermogenesis in brown adipose tissue. Conversely, administration of a beta3 adrenoceptor (beta3-AR) agonist decreases leptin mRNA expression and secretion, suggesting that leptin and sympathetic nervous system activity mediated through the beta3-AR comprise a negative-feedback loop. It has recently been proposed that a defect in the beta3-AR in humans may contribute to a resistance to the sympathetically mediated effects of leptin on thermogenesis and lipolysis, thus leading to obesity and type 2 diabetes mellitus. We thus hypothesized that the Trp64Arg variant in the human beta3-AR would be associated with elevated plasma leptin concentrations. We studied 101 healthy nondiabetic Pima Indians: 11 Arg64 homozygotes, 35 Trp64 homozygotes, and 55 heterozygotes. The fasting plasma leptin concentration as an absolute value or after adjustment for percent body fat and sex was not associated with the beta3-AR genotype. Thus, the data do not support an influence of the Trp64Arg variant on the plasma leptin concentration. PMID- 9867086 TI - A Trp 64 Arg mutation in the beta3-adrenergic receptor gene is not associated with moderate overweight in Japanese workers. AB - The aim of the present study was to investigate the trend for obesity and the relationship between Trp64Arg mutation in the beta3-adrenergic receptor gene and obesity-related phenotypes in Japanese workers. A total of 658 workers, 483 women and 175 men, responded to this survey. Genomic DNA was prepared from leukocytes, and DNA amplification by polymerase chain reaction was performed using 100 ng genomic DNA. Amplified fragments were digested with BstNI and analyzed by 3% agarose gel electrophoresis. The body mass index (BMI) increased from 21.1 +/- 2.6 to 22.0 +/- 3.0 kg/m2 for women and from 20.9 +/- 2.1 to 22.1 +/- 2.6 kg/m2 for men from 1982 to 1997. Body weight increased 63.3% for women and 67.4% for men over 15 years. The frequency of the Trp64Arg allele was 19.9%, and the frequency of heterozygote and homozygote carriers of the mutation was 32.9% and 3.4%, respectively. Changes in the BMI and body weight over 15 years from 1982 to 1997 did not differ significantly between subjects with or without the mutant allele in either women or men. The BMI was not significantly higher in subjects who were homozygous or heterozygous for the Trp64Arg mutation versus subjects who were homozygous for the normal allele. This mutation of the beta3-adrenergic receptor gene is not likely a major determinant of moderate obesity in the Japanese population. PMID- 9867087 TI - The emerging role of photodynamic therapy in the management of Barrett's oesophagus. PMID- 9867088 TI - Reconstitution of squamous epithelium in Barrett's oesophagus with endoscopic argon plasma coagulation: a prospective study. AB - BACKGROUND: Barrett's oesophagus is a premalignant condition. Recent reports have suggested that laser coagulation or photodynamic therapy combined with acid suppression may induce reconstitution of squamous mucosa. However, a high percentage of residual glands remain in cases treated with both techniques. Argon plasma coagulation (APC) appears to be an attractive alternative to other thermoablative techniques. The aim of this study was to investigate the reconstitution of squamous epithelium in Barrett's oesophagus after APC. METHODS: Fifteen patients with histologically proven Barrett's oesophagus were included in a prospective study. After base-line documentation by videotaping and biopsies, Barrett's epithelium was treated by repeated APC at intervals of 4-6 weeks until complete squamous restoration was achieved. All patients were kept under high dose proton pump inhibitor therapy. RESULTS: In 13 patients complete reconstitution of squamous epithelium was achieved. Buried glands after squamous restoration were detected transiently in only one case after the first session. As side effects seven patients had mild retrosternal discomfort. One patient reported severe retrosternal pain for 1 week. He then refused further APC sessions. Another patient was excluded because of noncompliance. During the follow-up period (6-13 months) recurrence of Barrett's epithelium was observed in one patient. CONCLUSIONS: APC is a suitable technique for achieving squamous restoration in Barrett's oesophagus. The rare occurrence of remaining buried glands may result from the homogeneous coagulation achieved by the ionized argon gas beam. PMID- 9867089 TI - Positive association between Helicobacter pylori infection and food allergy in children. AB - BACKGROUND: In children Helicobacter pylori has been involved as a pathogenetic factor in gastritis and duodenal ulcer and as a cofactor in protein-losing enteropathy, chronic diarrhoea, short stature, and gastritis lymphoproliferative disease. A subset of an H. pylori strain possesses an antigen, CagA, as a virulence factor. In the present study we determined anti-H. pylori IgG and anti CagA IgG titres in children with food allergy. METHODS: Ninety paediatric patients were studied: 30 with food allergy, 30 with atopic asthma, and 30 with inflammatory bowel disease. Anti-H. pylori IgG and anti-CagA IgG were determined in all children by means of a commercial enzyme immunoassay (ELISA). RESULTS: The anti-H. pylori IgG titre was significantly higher in allergic patients than in the other two groups. The anti-CagA IgG titre did not differ significantly between the patients. CONCLUSIONS: These findings show a positive association between H. pylori infection and food allergy in children. We hypothesize that virulence factors other than CagA may be involved in the pathogenesis of H. pylori infection in paediatric patients with food allergy. PMID- 9867090 TI - Detection of Helicobacter pylori gene by means of immunomagnetic separation-based polymerase chain reaction in feces. AB - BACKGROUND: Detection of Helicobacter pylori is usually performed by culture, polymerase chain reaction (PCR), histology, or urease test on gastric biopsy samples. Although methods based on feces are non-invasive, their sensitivity has been relatively low. In this study, to improve its sensitivity, immunomagnetic separation (IMS) was used as a pre-PCR step for direct detection of H. pylori in feces. METHODS: Fresh fecal samples were taken from 72 patients attending for endoscopy. Of these, 57 patients had a positive H. pylori status according to the results of culture, histology, and PCR on gastric biopsy samples. Anti-H. pylori antibody-sensitized immunomagnetic beads were used to concentrate the bacteria. PCR was then performed to detect the H. pylori urease A-encoding gene. RESULTS: Of the 57 H. pylori-positive patients, 35 (61.4%) had positive fecal samples by IMS-based PCR method. None of the 15 H. pylori-negative patients had positive fecal samples. The sensitivity of this method was 61.4%, and the specificity 100.0%. CONCLUSIONS: This study confirms that non-invasive diagnosis of H. pylori infection could be made from feces by using IMS-based PCR. PMID- 9867091 TI - Recurrence of Helicobacter pylori infection after eradication: incidence and variables influencing it. AB - BACKGROUND: Our aim was to study the incidence of Helicobacter pylori recurrence in our country and to assess the different variables that might influence it. METHODS: We studied prospectively 331 duodenal ulcer patients (mean age, 48 +/- 14 years, 71% male) in whom H. pylori had been eradicated. Several therapies were used, classified as low-efficacy (omeprazole + amoxycillin, 32% eradication rate; omeprazole + amoxycillin + metronidazole, 56%) and high-efficacy therapies (omeprazole + clarithromycin + amoxycillin or metronidazole, 88%; bismuth triple therapy, 77%). One month after completion of therapy an endoscopy with biopsies and/or 13C-urea breath test was performed. A breath test was carried out again at 6 months, 1 year, and 2 years, to study H. pylori recurrences. Endoscopy (with biopsies) was performed only to confirm recurrences. Multiple logistic regression analysis was used. Differences between Kaplan-Meier curves were evaluated with the log-rank test. RESULTS: Sixty-seven patients were followed up for 6 months, 136 for 1 year, and 128 for 2 years, giving 425 patient-years of follow-up. A total of 18 H. pylori recurrences was observed (12 at 6 months, 4 at 1 year, and 2 after 2 years), yielding a yearly recurrence of 4.2% patient-years(-1). The respective risk of H. pylori recurrence for each period was 3.6% (95% confidence interval (CI), 2.1%-6.2%), 1.5% (0.6%-3.8%), and 1.5% (0.4%-5.5%). The probability of being H. pylori-negative at 6 months, 1 year, and 2 years was, respectively, 96.4% (94.4%-98.4%), 94.9% (92.5%-97.4%), and 93.4% (90.3%-96.6%). Duodenal ulcer was found in half of the reinfected patients. The recurrence rate at 6 months was 10.3% (5.7%-18%) in patients <40 years old and only 0.85% (0.2% 3.1%) in those > or =40 years old (P = 0.0002). Of the patients who became reinfected at 6 months 27% (6%-61%) had delta breath test values between 3 per thousand and 5 per thousand 1 month after therapy, whereas only 4.6% (2.7%-7.7%) of non-reinfected patients had delta after eradication > or =3 per thousand (P = 0.0097). H. pylori recurrence at 6 months was 8.2% (4.5%-15%) in patients previously treated with low-efficacy therapies and only 1.7% (0.7%-4.3%) when high efficacy therapies were used (P = 0.0098). In the multivariate analysis age (odds ratio (OR), 0.9; 95% CI, 0.8-0.96; P = 0.0008), the delta breath test value after therapy (OR, 2.2; CI, 1.2-4.1; P = 0.0076), and therapy regimen (OR, 6.4; CI, 1.5-27; P = 0.0109) were the only variables that correlated with H. pylori recurrence at 6 months. Differences were observed when Kaplan-Meier curves were compared, depending on age (<40 or > or =40 years; P = 0.0054), breath test value (delta) 1 month after therapy (<3 or > or =3 per thousand; P = 0.0089), and therapy regimen (high or low efficacy; P = 0.0006). CONCLUSIONS: Risk of post eradication H. pylori recurrence is higher during the first 6 months, which suggests that most recurrences during this period are recrudescences and not true reinfections. Patients who have H. pylori recurrence tend to be younger and have higher delta 13C-urea breath test values after therapy, which suggests that a 'negative' value between 3 per thousand and 5 per thousand needs to be confirmed. Recurrence of H. pylori is more frequent in patients treated with low-efficacy therapies but is exceptional when high-efficacy therapies are used, in which case post-therapy eradication can be safely confirmed at 4 weeks. Finally, recurrence of H. pylori is clinically relevant, as ulcer recurrence is observed in a considerable proportion of these patients. PMID- 9867092 TI - Vacuolating toxin gene polymorphism among Helicobacter pylori clinical isolates and its association with m1, m2, or chimeric vacA middle types. AB - BACKGROUND: Helicobacter pylori vacuolating cytotoxin encoded by vacA plays an essential role in H. pylori-related pathogenesis. Specific vacA alleles are believed to be associated with increased virulence. Association among vacA polymorphism, vacA middle genotypes, and various H. pylori-related diseases was thus investigated. METHODS: Eighty-nine isolates from patients with various gastrointestinal diseases were examined for restriction fragment length polymorphism (RFLP) of the 2.0-kb polymerase chain reaction-amplified vacA middle region. Further genetic heterogeneity was assessed with ureA-ureB RFLP. RESULTS: Twenty-eight distinct vacA RFLPs were seen among 89 isolates. Each pattern was associated with one specific vacA middle genotype. The association of specific RFLPs with certain clinical manifestations was noted among six common groups. Further RFLP analysis of the 2.4-kb ureA-ureB segment from isolates in four popular vacA RFLPs showed high genetic variation. CONCLUSIONS: The vacA genetic polymorphism may be associated with different gastrointestinal diseases. PMID- 9867093 TI - Fructose-sorbitol malabsorption and symptom provocation in irritable bowel syndrome: relationship to enteric hypersensitivity and dysmotility. AB - BACKGROUND: Fructose-sorbitol (F-S) mixtures can provoke symptoms in irritable bowel syndrome (IBS) patients, and a proportion of IBS patients also have enteric hypersensitivity to distension. We hypothesized, therefore, that sugar malabsorption and fermentation to produce hydrogen gas may provoke symptoms to a greater extent in IBS patients hypersensitive to distension than in those patients without such hypersensitivity. Our aims were therefore to compare, in IBS patients, symptoms and breath hydrogen responses after F-S, on the basis of jejunal sensitivity and jejunal motor function. METHODS: Fifteen female IBS patients (44 +/- 15 years) underwent, on separate occasions, 3-h breath hydrogen analyses after ingesting 10 g lactulose and 25 g fructose with 5 g sorbitol. Jejunal sensitivity and motor function were determined by balloon distension and 24-h manometry studies, respectively. Cumulative symptom scores and breath hydrogen production were analysed on the basis of the presence or absence of jejunal hypersensitivity and dysmotility. RESULTS: Four and seven patients had jejunal hypersensitivity for initial perception and pain, respectively. Eleven, nine, and nine patients had jejunal dysmotility for fasting phase 3, phase 2, and fed motor activity, respectively. Of the patients with symptom provocation after F-S (n = 8 within 3 h, n = 12 within 12 h) or with F-S malabsorption (n = 10), the relative proportion did not differ on the basis of the presence or absence of jejunal hypersensitivity or of motor dysfunction. Symptom scores and hydrogen production also were not different in these subgroups. CONCLUSIONS: Although carbohydrate malabsorption can provoke symptoms in some IBS patients, there is no consistent association between such a phenomenon and the presence of either jejunal hypersensitivity or dysmotility. PMID- 9867094 TI - Electrogastrographic study of the effect of transcatheter arterial chemoembolization on gastric myoelectric activity. AB - BACKGROUND: Transcatheter arterial chemoembolization (TACE) of the hepatic artery is frequently used in the treatment of inoperable hepatocellular carcinoma (HCC). TACE causes not only effective tumor tissue necrosis in patients with hepatoma but also adverse effects on extrahepatic abdominal organs. There are no published reports on the effect of TACE on the gastric myoelectric activity. In this study, using cutaneous electrogastrography (EGG), we evaluated the effect of TACE on gastric myoelectric activity in patients with HCC. METHODS: A total of 27 patients (24 men and 3 women, aged 22 to 78 years) with hepatoma, admitted for TACE, were included in this study. Furthermore, 28 patients (24 men and 4 women, aged 26 to 75 years), admitted for diagnostic angiography of the liver, served as the control group. Cutaneous EGG was performed before and after TACE or angiography. RESULTS: In the TACE group there were significant changes in dominant frequency (DF) and percentages of DF in the defined normal range, bradygastric range, and tachygastric range on post-meal EGG. On fasting EGG, only the dominant frequency and percentages of DF in the bradygastric range changed significantly. However, there was no correlation between the occurrence of nausea/vomiting and the degree of change in the EGG variables, during both fasting and postprandial states. In the control group there were no significant differences in EGG variables before and after angiography. CONCLUSIONS: TACE can affect gastric myoelectric activity in HCC patients. Nevertheless, the relationship between changes in myoelectric activity and the occurrence of gastrointestinal symptoms needs further investigation. PMID- 9867095 TI - Serosal but not mucosal endotoxin exposure increases intestinal permeability in vitro in the rat. AB - BACKGROUND: Microbial endotoxins are normally present in the gut, usually without apparent harmful effects, whereas systemically administered endotoxin impairs the mucosal barrier function. Our aim was to investigate whether in vitro exposure to bacterial lipopolysaccharide (LPS) could affect the intestinal barrier properties of the rat small intestine. METHODS: Small-intestinal segments from rats were mounted in Ussing diffusion chambers, and the mucosal to serosal permeation of the marker molecules bovine serum albumin (BSA) and 51Cr ethylenediaminetetraacetic acid (EDTA) was measured after addition of LPS to the mucosal or serosal side. RESULTS: Mucosal exposure to LPS (0.01, 0.05, 0.25 mg/ml) had no effects on the permeation of BSA and 51Cr-EDTA, whereas when added to the serosal side at 0.05 or 0.25 mg/ml, LPS increased the marker permeation. CONCLUSION: Serosal LPS exposure in vitro increased the intestinal permeability to the different-sized markers, whereas mucosal LPS did not, indicating that the mechanisms leading to intestinal barrier impairment can be initiated in the intestinal wall itself. PMID- 9867096 TI - Significance of a preserved colon for parenteral energy requirements in patients receiving home parenteral nutrition. AB - BACKGROUND: Colonic digestion has been reported to salvage up to 3-4 MJ/day in short-bowel patients (approximately 50% of the daily requirements). METHODS: A cross-sectional study of the parenteral support given to a total cohort of 73 patients receiving home parenteral nutrition (HPN) in Denmark on 31 December 1995 was performed to evaluate the significance of a preserved colon for the prevalence of short-bowel patients in need of HPN and the requirements of energy given as HPN. The number of HPN patients with a substantial remnant colon (> or =50%) was compared with the number with no colonic function (0%) in subgroups of patients with remnant small bowels of <100 cm (group 1), 100-200 cm (group 2), and >200 cm (group 3). Patients with pseudo-obstruction (n = 8) and patients with <50% colon remaining (n = 11) were excluded from the analysis. The impact of a substantial preserved colonic function (> or =50%) on the parenteral energy requirements was evaluated in patients with comparable lengths of small bowel. RESULTS: Twelve and 8 patients in group 1 had no colon and > or =50% colon in function, respectively, in contrast to patients in group 2, in which 21 had no colon in function and only 2 had >50% in function (Fisher's exact test, P = 0.028). In patients in group 1 the need for parenteral energy in percentage of basal energy expenditure (HPN/BEE%, mean +/- standard deviation) was 110 +/- 31% in patients with no colon and 59 +/- 31% in patients with a preserved colon (P = 0.001). In patients without a colon in groups 2 and 3 the HPN/BEE% was 58 +/- 45% and 33 +/- 47%, respectively. CONCLUSIONS: Preservation of a substantial colonic function (> or =50%) is rare in patients in need of HPN with >100 cm remnant small bowel and results in a reduction in the parenteral energy requirements of approximately 3 MJ/day (51% of BEE) in patients with <100 cm small bowel. These data reinforce the reports of the colon as an energy-salvaging organ (approximately 3-4 MJ/day), which makes HPN unnecessary in most patients in whom small-bowel length is sufficient (>100 cm) to absorb another 3-4 MJ/day. PMID- 9867097 TI - Bone mass and metabolism in Whipple's disease: the role of hypogonadism. AB - BACKGROUND: Whipple's disease, like other malabsorption syndromes, ought to predispose to osteopenia. We therefore evaluated bone mass and mineral metabolism in a cohort of patients with this condition. METHODS: Twelve male patients with Whipple's disease and 36 male age-matched healthy subjects took part in the study. None of the patients complained of diarrhea at the time of the study. Bone mineral density at the lumbar and femoral level and serum levels of indices of bone and mineral metabolism and of gonadal function were measured. RESULTS: Bone mineral density at the total femur and femoral neck were significantly lower in patients with Whipple's disease than in healthy volunteers, whereas no significant difference was found at the lumbar level. In patients with Whipple's disease serum levels of type-I collagen teleopeptide (ICTP) and sex-hormone binding globulin were significantly higher, whereas serum levels of testosterone and luteinizing hormone were significantly lower than in healthy volunteers. Moreover, testosterone correlated significantly (P < 0.05) with lumbar bone mineral density (r(s) = 0.64) and serum ICTP levels (r(s) = -0.63). CONCLUSIONS: In patients with previously treated Whipple's disease and without any current symptoms of malabsorption, bone loss is generally moderate and linked to the presence of hypogonadism. PMID- 9867098 TI - Presence of high levels of non-degraded gliadin in breast milk from healthy mothers. AB - BACKGROUND: Secretion of dietary antigens into breast milk has been extensively documented. The presence of these antigens is of relevance because they could be involved in the modulation of the immune response in neonates. The objective of this study is to determine the gliadin concentration in milk, colostrum, and serum samples from healthy lactating mothers on a normal diet. Gliadin levels in milk samples from a group of six mothers after a brief period of gluten restriction were also determined. The molecular weight of secreted gliadins was also analysed. METHODS: Gliadin concentration was determined with a highly sensitive competitive enzyme-linked immunosorbent assay, modified so as to eliminate anti-gliadin antibody interference. The level of gliadin/IgA anti gliadin immune complexes in milk, colostrum, and serum samples was determined. RESULTS: Gliadin was detected in all 49 milk samples. Its concentration varied between 5 and 1200 ng/ml (mean, 178 ng/ml). In colostrum (n = 14) gliadin levels were higher (range, 28-9000 ng/ml; mean, 883 ng/ml), not being detectable in one case. Gliadin was detectable in 14 of 31 serum samples, in which levels were lower than in milk and colostrum samples (mean, 41 ng/ml). Neither a correlation between gliadin levels in milk, colostrum, and serum samples from the same subject nor a relation between gluten intake and gliadin concentration in milk samples from six subjects under a 3-day gluten-free diet could be found. Higher levels of immune complexes were observed in colostrum samples than in milk and serum samples. No correlation was detected between gliadin concentration and the level of immune complexes. The analysis of milk and colostrum samples by immunoblotting showed bands of immunoreactive gliadin presenting Mr similar to those of native proteins from wheat extracts. CONCLUSIONS: Very high levels of gliadin were detected in milk samples from healthy mothers on an unrestricted diet. Gliadin levels were higher than those reported for dietary antigens in other studies. Breast milk contained non-degraded gliadins and gliadin/anti gliadin IgA immune complexes. PMID- 9867099 TI - Active Crohn's disease and ulcerative colitis evaluated by low-field magnetic resonance imaging. AB - BACKGROUND: Our aim was to evaluate low-field magnetic resonance imaging (MRI) in the assessment of disease extension and activity in inflammatory bowel disease. METHODS: Nineteen patients with Crohn's disease (CD), 8 with ulcerative colitis (UC), and 5 healthy controls (HC) were examined using MRI (0.1 T) before and after intravenously administered gadodiamide and glucagon. MRI images were evaluated in a blinded fashion and compared with findings at endoscopy, double contrast barium enema, small-bowel follow-through, and surgery. RESULTS: Comparisons of diseased with both non-diseased bowel segments and segments from HC showed significant differences for both CD and UC with regard to signal intensity on T2-weighted (SI(T2)) images and post-contrast increment of signal intensity on T1-weighted images (%SI(T1). Agreements with regard to disease extension in CD between MRI and other examinations were 97%, underestimating the extension in two patients. For SI(T2) in CD a cut-off value of 1.0 showed a predictive value of a positive finding (PVpos) = 1.0 and a predictive value of a negative finding (PVneg) = 0.96. For %SI(T1) in CD a cut-off value of 15.0% showed values of PVpos = 0.95 and PVneg = 0.92. Agreements between MRI and conventional methods (disease extension) in UC was 87.5%. Extension was underestimated in two patients and overestimated in two patients as compared with barium enemas. Values of PVpos were 1.0 (SI(T2) >1.0) and 1.0 (%SI(T1) >15.0%), respectively, with corresponding values of PVneg being 0.94 and 0.94. CONCLUSION: Low-field MRI seems a promising non-invasive, non-radiating method in the evaluation of inflammatory bowel disease. PMID- 9867100 TI - The accuracy of abdominal ultrasound in the assessment of bowel disorders. AB - BACKGROUND: Little is known about the sensitivity, specificity, and predictive values of transabdominal ultrasonographic (US) findings in a teaching hospital setting. METHODS: We carried out a prospective study including 227 patients with symptoms suggestive of inflammatory bowel disorder. The Picker 9200 CS equipment (5-mHz curved-array probe) was used to obtain bowel images. Gastrointestinal endoscopy, enteroclysis, bowel enema, computed tomography scan, or bowel surgery was used as reference. RESULTS: Of 227 patients, 168 had pathologic findings of the bowel as final diagnosis. The overall sensitivity of US was 76%, whereas the positive predictive value was 98%. Overall specificity was 95%. The negative predictive value for bowel disorders was only 58%, since US missed pathologic findings in 48 patients. Subgroup analysis showed a sensitivity of 84% for Crohn's disease, 66% for ulcerative colitis, 46% for bowel tumors, and 60% for diverticulitis. Topographic comparisons showed that US detected inflammatory bowel-wall alterations preferentially in the terminal ileum and colon, whereas abnormalities in the duodenum, jejunum, and rectum were frequently missed (sensitivity, 10%-20%). CONCLUSIONS: Positive US findings are useful for the diagnosis of bowel processes. US is highly predictive albeit not disease specific. Negative US examinations, however, do not exclude pathologic bowel processes. A topographic location of pathologic US findings is mostly confined to the colon. PMID- 9867101 TI - Lack of influence of hepatitis G virus infection on alcohol-related hepatic lesions. AB - BACKGROUND: The purposes of this study were to analyse the prevalence and histologic impact of hepatitis G virus (HGV), a newly discovered virus, in alcoholic patients, a population known to be at risk for viral hepatitis. METHODS: One hundred and thirty-nine consecutive alcoholics admitted to our liver unit (106 men and 33 women; mean age, 47.1 +/- 10.9 years) were included in the study. All patients had consumed more than 60 g of ethanol per day for at least 1 year. One hundred healthy blood donors constituted a control group. Antibodies to HGV E2 protein and HGV-RNA testing by reverse transcription-polymerase chain reaction (RT-PCR) with primers derived from the NS5 coding region were performed in all serum samples. RESULTS: A significantly higher seroprevalence of anti-E2 antibodies was observed in alcoholic patients than in healthy blood donors (41 (29.5%) versus 8 (8%); P < 0.0001). Moreover, the prevalence of HGV-RNA was significantly higher in alcoholic patients (13 (9.3%) versus 1 (1%); P = 0.01). HGV-RNA and anti-HGV antibodies were never detected simultaneously. HGV viraemia was not associated with an increased risk of cirrhosis or hepatocarcinoma in alcoholic subjects. CONCLUSIONS: Our study reports a high prevalence of HGV in alcoholic patients. HGV infection does not modify or aggravate the course of alcoholic liver disease. PMID- 9867102 TI - Prognostic value of progressive decrease in serum cholesterol in predicting survival in Child-Pugh C viral cirrhosis. AB - BACKGROUND: The identification of cirrhotic patients with low life expectancy is an open clinical problem. Hypocholesterolemia is frequently found in severe chronic hepatic insufficiency because the liver is the most active site of cholesterol metabolism, but poor information is available on its precise prognostic value. We evaluated the prognostic role of hypocholesterolemia in patients with advanced liver cirrhosis. METHODS: Serial serum cholesterol concentrations of 34 patients with virus-induced cirrhosis, from the first appearance of Child-Pugh class C to death, were considered. To compare survival functions, we established three base-line cholesterol cut-off points (150, 125, and 100 mg/dl) and stratified patients into groups A and B, with base-line cholesterol levels lower and higher than each cut-off value, respectively. RESULTS: Cholesterolemia decreased progressively in all patients. At the 100 mg/dl cut-off point all group-A patients died within 17 months, whereas 75% of group-B patients were alive at 24 months (P < 0.0001). Moreover, cholesterolemia was significantly correlated with cholinesterase, indirect bilirubin, and total bilirubin at entry time and immediately before death. No correlation was observed between cholesterol and these variables when stratified for the Child-Pugh score. CONCLUSIONS: Base-line serum cholesterol levels lower than 100 mg/dl identify a subgroup of Child-C cirrhotic patients with high mortality risk within a 2-year follow-up. The prognostic importance of cholesterolemia may also be deduced by the significant correlation with other well-established indicators of survival. PMID- 9867103 TI - Prevalence of gallstone disease in a Swedish population sample. Relations to occupation, childbirth, health status, life style, medications, and blood lipids. AB - BACKGROUND: There are only a few Swedish studies on the prevalence of gallstone disease in selected age groups, and none including possible risk factors. METHODS: Of a population sample of 1200 individuals (age, 35-85 years), 857 participated in the study. The study subjects were asked to answer a questionnaire about potential risk factors (occupation, childbirth, life style, and so forth), symptoms, and quality of life. Cholecystectomy had previously been done in 115 subjects, leaving 742 for ultrasound examination of the gallbladder. RESULTS: The prevalence of gallstone disease increased with age, and at 75 years or more, 53% of the women and 32% of the men either had gallstones or had previously undergone cholecystectomy (32% and 13%, respectively). When comparing subjects with and without gallstones, there were no differences with regard to any variable, including blood lipid levels. The odds ratio of previous cholecystectomy was increased in subjects with an occupation requiring no specific education and reduced in subjects using wine or spirits every week. The odds ratio of abdominal pain was increased after previous cholecystectomy. Women in this group also experienced a lower quality of life. CONCLUSIONS: The age and sex distribution of gallstone disease was in the order of the magnitude seen in other Scandinavian countries. None of the studied variables differed between subjects with and without gallstones. Subjects previously operated on with cholecystectomy did worse with regard to symptoms and quality of life. PMID- 9867105 TI - Intragastric pH and Helicobacter pylori treatment with proton pump inhibitors combined with amoxycillin. PMID- 9867104 TI - Immunohistochemical analysis of a case of gastritis cystica profunda associated with carcinoma development. AB - We report a rare case of gastritis cystica profunda (GCP) accompanied by carcinoma that developed in a 51-year-old Japanese man without antecedent gastric surgery. The polypoid tumor was located in the upper body of the resected stomach. Histologically, it was characterized by herniation of surface epithelium and cystic glands in the submucosa, muscularis propria, and subserosa. Marked chronic atrophic gastritis was found throughout the stomach, and dysplastic epithelia and a few adenocarcinoma cells were found in the deeper parts of the GCP. The Ki-67, p53, and p21WAF1/CIP1 labeling indices for the deeper part of the GCP were higher than those for the superficial parts or the surrounding mucosa, suggesting that both epithelial cell proliferation and p53-dependent p21WAF1/CIP1 expression in DNA-damaged cells, which might be associated with gastritis, are enhanced in line with penetration of glands. The underlying mechanisms might be linked in a chain of factors leading to malignancy. PMID- 9867106 TI - Effect of cytokines on acid secretion and gastrin secretion in Helicobacter pylori infection and aspirin-induced gastritis. PMID- 9867108 TI - Agonists and antagonists of regulatory peptides as tools to study regulation of pancreatic exocrine secretion, cell proliferation and gene expression. AB - For more than two decades, our research group has been studying the pancreatic actions of three groups of regulatory peptides: members of the cholecystokinin/gastrin family, bombesin-like peptides and somatostatin. Investigating these peptides, our work has focused on three particularly interesting aspects: peptidergic regulation of pancreatic enzyme secretion and growth in adult rats, peptidergic control of pancreatic enzyme secretion and growth during postnatal development in rats, and peptidergic regulation of proliferation and differential gene expression in pancreatic adenocarcinoma cells. Our data confirmed that the control of the exocrine function of the pancreas is complex, and that it involves peptides such as the cholecystokinin/gastrin-like peptides, bombesin-like peptides and somatostatin. In these investigations, it became evident that selective peptide receptor agonists, antagonists and monoclonal antibodies raised against peptides are useful tools to identify the role of these bioactive peptides in pancreatic exocrine secretion and cell proliferation. PMID- 9867107 TI - Immunohistochemical findings in the jejunal mucosa of patients with coeliac disease. AB - In the pathogenesis of coeliac disease, disturbed immunological processes play a basic role. This is also proved by immunohistochemical findings in the jejunal mucosa of patients with this disorder, which are discussed in this review. Intraepithelial lymphocytes are increased in number in active coeliac disease, most being of the CD8 subpopulation. The counts of lamina propria lymphocytes and the relative numbers of their subsets are comparable to those in healthy controls, but several studies have indicated that the densities of IgA, IgM and IgG immunoglobulin-containing cells in the lamina propria are increased in untreated coeliac disease. Activated T cells can also be found in the lamina propria of patients on gluten-containing diets. In the crypt epithelial cells of coeliac patients on normal diets, strong expression of HLA Class II antigens can be observed. All of these changes are normalized on a gluten-free diet. It was recently discovered by the authors and others that one of the hallmarks of coeliac disease, both active and treated, is an increase in number and proportion of gamma/delta intraepithelial T lymphocytes. The permanent increase of gamma/delta T cells may facilitate identification of a coeliac patient also when an intestinal biopsy is taken on a gluten-free diet and other morphologic signs are normalized. The detection of elevated numbers of intraepithelial gamma/delta T cells is also an indispensable help in the diagnosis of latent coeliac disease. PMID- 9867109 TI - Gastric blood flow and acid secretion: problems of the methodology and regulatory mechanisms. AB - Methods for proper measurement of gastric mucosal blood flow (MBF) in animal experiments or in clinical studies are critically reviewed. It can be stated that none of the techniques hitherto published fulfill all the criteria for proper measurement of the mucosal blood flow and concomitant acid production in the stomach. Some give an estimate of local blood flow changes (e.g. H2 gas clearance, laser Doppler flowmetry) in man without the possibility of simultaneous acid measurement. The 99mTc-MAP clearance technique developed by the authors reflects changes in both parameters and may also be used in clinical studies, when limitations of the method are taken into consideration. As a rule we assume that acid production and mucosal blood flow may change independently. The effects of various bioactive substances and drugs on both parameters are analysed. It seems that many contradictory findings have arisen because of differences in technique and experimental animals used. PMID- 9867110 TI - Progress in the management and treatment of infected pancreatic necrosis. AB - Infected pancreatic necrosis and sepsis are the leading causes of mortality in necrotizing pancreatitis. A review has been undertaken of the results of the past two decades relating to different surgical treatments of infected pancreatic necrosis. During the period 1978-85, the surgical treatment of necrotizing pancreatitis and its complications in our department consisted of the 'conventional' therapy (resection of the involved pancreatic tissue, or necrosectomy and drainage) in 61 patients, with a mortality rate of 36% (22 patients died). Since 1986, we have performed necrosectomy and other surgical interventions combined with continuous widespread lavage in 142 patients with infected pancreatic necrosis. The overall mortality decreased significantly to 6.3% (9 patients died). This result was achieved by means of aggressive surgical treatment, continuous, prolonged washing and suction drainage and supportive therapy, including immunonutrition, modifying the cytokine production and adequate antibiotic and antifungal medication. This surgical strategy provides the possibility for recovery in cases of necrotizing pancreatitis with septic complications. PMID- 9867111 TI - Oxidative stress in the liver and biliary tract diseases. AB - Oxidative stress plays an important role in the pathogenesis of toxic liver diseases and of other hepatic alterations. We summarize the pathomechanism of free radical reactions in liver diseases and the results of experimental and clinical observations. Most of the hepatoprotective drugs belong in the group of free-radical scavengers, their mechanism of action involving membrane stabilization, neutralization of free radicals and immunomodulation. We demonstrate the effects of the different drugs used in the therapy of liver diseases in animal experiments and in human clinicopharmacological studies. The scavenger effect of these drugs has been demonstrated in the subcellular fractions of liver cells in animal experiments. In vitro incubation with some hepatoprotective drugs inhibit lectin-induced lymphocyte transformation while others decrease the antibody-dependent, spontaneous, and lectin-induced lymphocytotoxicity. Dihydroquinolin-type antioxidants and silymarin enhanced the superoxide dismutase activity of erythrocytes and lymphocytes. In addition, in a 6-month double-blind clinical trial of patients with chronic alcoholic liver disease, we studied the effects of silymarin therapy on liver function tests, on the parameters characterizing the oxidative stress and immune reaction, on serum procollagen III peptide level, and on liver histology. A wide range of methods was used. The silymarin preparate corrected the altered immune reaction and the decreased superoxide-dismutase activity of erythrocytes and lymphocytes in patients with alcoholic liver cirrhosis. The results indicate that these drugs exert hepatoprotective activity and can improve liver functions in alcoholic patients and in toxic liver diseases. We found a correlation between the bilirubin concentration and lipid peroxidation in cases with toxic liver and biliary tract diseases, and assume that there are two kinds of bilirubin, an antioxidant and a prooxidant form, on the basis of diene conjugates in the bile. PMID- 9867112 TI - Increased interleukin-6 levels, interleukin-6 receptor and gp130 expression in peripheral lymphocytes of patients with inflammatory bowel disease. AB - Interleukin-6 is one of the most well-characterized cytokines with pleiotropic properties. Besides its B-lymphocyte activation role in hematopoiesis, interleukin-6 plays a central role in regulation of systemic inflammation. Interleukin-6 binds to receptors on target cells (such as hepatocytes and lymphocytes), consisting of an 80 kDa binding chain and gp130, a polypeptide responsible for signal transduction. In addition to the detection of elevated amounts of interleukin-6 in the blood, gene expression (mRNA) of subunits of the interleukin-6 receptor complex have also been studied by examining the reverse transcriptase polymerase chain reaction on peripheral lymphocytes from patients with characteristic radiological symptoms suffering from Crohn's disease. Our data show significantly elevated gene expression both of the 80 kDa interleukin-6 binding chain and gp130. These results suggest that enhancement of the expression of the constituents of interleukin-6 and the interleukin-6 receptor system plays a relevant role in systemic inflammation in inflammatory bowel disease. PMID- 9867113 TI - Experimental and human liver fibrogenesis. AB - In this work, we provide an overview of our results obtained by studying the role of transforming growth factor beta1 and proteoglycans in liver fibrogenesis. It has been found that transforming growth factor beta1 is one of the most important stimulators of extracellular matrix synthesis in the liver. In chronic liver injury, desmin-positive non-parenchymal liver cells expressed transforming growth factor beta1. The extracellular localization of the growth factor correlated well with types I, III and IV procollagen-alpha, which were detected in the fibrous septa of chronically injured livers. A similar distribution pattern was observed in human specimens. To identify the role of transforming growth factor beta1 in liver extracellular matrix protein synthesis, transforming growth factor beta1 positive transgenic mice were generated. Animals expressing the growth factor in their liver showed spontaneous liver fibrosis. Proteoglycans also participate in fibrogenesis. The majority of liver-specific heparan sulfate proteoglycans, such as syndecan-1 and fibroglycan, are produced by hepatocytes. The extracellular matrix proteoglycans decorin and perlecan are synthesized by non-parenchymal liver cells. The amount of the latter is very low in normal liver, but increases dramatically in liver fibrosis. The effect of regulatory factors on liver proteoglycans seems to be cell type-specific. In contrast to previous observations, elevated amounts of decorin did not inhibit the action of transforming growth factor beta1 in the liver. PMID- 9867114 TI - Histological evaluation of the response to interferon-alpha therapy in chronic hepatitis C. AB - BACKGROUND AND AIMS: The response rate of interferon-alpha (IFN-alpha), recently introduced in the treatment of chronic hepatitis C, is merely 25-50%. The aims of this follow-up study were to compare the efficacy of 6 and 12-month IFN-alpha treatment via liver biopsy scores and to evaluate the correlation with the biochemical response. PATIENTS AND METHODS: Twenty chronic hepatitis C patients were studied; 10 received IFN-alpha therapy for 6 months and 10 for 12 months. Liver biopsy material was taken before and after therapy. RESULTS: There was a significant serum alanine aminotransferase (ALT) level improvement in both groups, but a significant histological improvement in necroinflammatory activity (grade) only in the 12-month group. The Chevallier stage scores demonstrated a significant progression in both groups. CONCLUSIONS: Twelve-month IFN-alpha treatment affords a better response in the liver histology grade and serum ALT level, but does not influence the staging; a normal ALT does not guarantee hepatitis inactivity. Liver biopsies appear indispensable for monitoring. PMID- 9867115 TI - Deleterious action of vasopressin in gastroduodenal ulceration: experimental and clinical observations. AB - Vasopressin, released from the posterior pituitary and from the vascular endothelium, can cause vasoconstriction and provoke platelet aggregation, leading to an impaired tissue blood supply. In humans with pituitary diabetes insipidus the central release of vasopressin is diminished, and in the Brattleboro homozygous rat there is congenitally no synthesis of this hormone. The gastroduodenal intramucosal vasopressin level is elevated in normal rats following various acute ulcerogenic challenges (after ethanol, reserpine, indomethacin, cold-restraint stress, endotoxin shock and hemorrhagic shock), and vasopressin-deficient rats are less sensitive to these stimuli. In a hospital- and population-based case-control, age-matched retrospective study, the incidence of human gastroduodenal ulceration is significantly higher in the normal population (in whom the release of vasopressin is presumed to be intact) than in the vasopressin-deficient one (central diabetes insipidus patients). In conclusion, endogenous vasopressin plays an aggressive role in development of gastroduodenal ulceration, especially that related to stress. PMID- 9867117 TI - Humoral mechanisms and clinical aspects of biliary tract motility. AB - This review is intended to summarize current information on neurohumoral regulation of the gallbladder and sphincter of Oddi motility under both physiological and pathological circumstances with emphasis on Hungarian contributions to today's knowledge. The mechanism of action of neurohumoral agents that interact on these segments of the biliary tract, and the explored details of the stimulation-contraction/relaxation coupling process of these substances, will be discussed. A modified classification of biliary tract motility disorders with new diagnostic and therapeutic approaches will also be provided. This information will aid understanding of the pathogenesis of motor disorders of the gallbladder and sphincter of Oddi, and will indicate possibilities for pharmacological exploitation in the treatment of diseases resulting from biliary tract motility abnormalities. PMID- 9867116 TI - Clinical significance and application of anorectal physiology. AB - During the last decade, interest in anorectal disorders has increased continuously. This review presents the results of Hungarian authors' contributions to a better understanding of anorectal physiology over the last 10 years. It has been demonstrated that the surgical technique of pelvic floor reconstruction can be refined and surgical complication rates reduced through the analysis of anal basal pressure components and exploration of internal anal sphincter function. Objective assessment of anal continence (distinguishing continent from incontinent patients) is a consistent problem in clinical practice. The balloon-retaining test, a special application of manometry, facilitates evaluation of anorectal function in incontinent patients. Constipation is an extremely common symptom. Surgical treatment of these ailments should not be regarded as a first-choice therapy. Disordered defecation due to anismus can be identified by defecometry, and can eventually be treated by biofeedback training. The data presented here demonstrate the enormous impact of sophisticated diagnostic techniques on the therapeutic options in treatment of anorectal diseases. PMID- 9867118 TI - Human and experimental hepatocarcinogenesis. AB - Human liver cancer is increasing worldwide, including in Hungary. The detection of liver tumors in premalignant or early malignant states is essential for successful treatment. MC-29 virus-induced chicken hepatoma and rodent, fish and monkey models for chemical hepatocarcinogenesis were studied and compared to humans. Changes in phenotypic enzyme alterations and in the expression of certain oncogens and growth factors characterize the experimentally induced hepatomas, and might also be characteristic of human premalignant and malignant focal liver lesions. Fish hepatocarcinogenesis is useful for studying compounds in environmental pollution. Increased expression of transforming growth factor a can be observed both in experimental and human liver tumors. Increased tumor incidence was detected in transgene mice containing both transforming growth factor alpha and c-myc genes. Animal models of hepatocarcinogenesis help to understand the development of liver tumors. Methods applied in studies using those models are useful in the study of premalignant and malignant human liver lesions. PMID- 9867119 TI - Pain relief and functional recovery after endoscopic interventions for chronic pancreatitis. AB - BACKGROUND: Pain and functional deterioration in chronic pancreatitis is multifactorial. Early surgery in non-alcoholic patients with mild to moderate chronic pancreatitis can relieve pain and prevent progression of pancreatic insufficiency for some time, but the good results are only short term. Endoscopic intervention can relieve pain and recover pancreatic function without surgery. METHODS AND RESULTS: To achieve the burned out state of chronic pancreatitis, occlusion of the pancreatic duct was first attempted by our team with Ethibloc at ERCP. Temporary obstruction of the pancreatic duct did not result in a long lasting symptom and relapse-free situation because of early recovery of pancreatic function. On the contrary, endoscopic simple and double papillotomy, pancreatic drainage with citrate lavage, biliary endoprosthesis with multiple stents and endoscopic decompression of pseudocysts with or without jejunal feeding resulted in pain-free patients for a considerable time and in several cases significant functional recovery occurred. In cases where pain remained, percutaneous celiac plexus block with long-lasting steroids can be applied and only if all of these treatments fail should surgery be recommended. CONCLUSION: Endoscopic intervention can successfully substitute for surgery for chronic pancreatitis in individual cases. PMID- 9867120 TI - Hepatitis C virus infection: pathogenesis, diagnosis and treatment. AB - We discuss the basic questions concerning hepatitis C virus (HCV) infection and describe the molecular biological and epidemiological features of the causative agent HCV, as well as the pathogenetic mechanisms of the HCV-related hepatic and non-hepatic diseases. We then summarize the diagnostic and therapeutic procedures in accordance with the recent literature. All the problems are discussed from the point of view of Hungarian hepatologists who have given an account of the experiences and possibilities concerning these fields in Hungary. PMID- 9867121 TI - Somatostatin and the gastrointestinal tract. AB - Somatostatin has various significant effects on the gastrointestinal tract: it inhibits the release of several gastrointestinal hormones, blocks the exocrine function of the stomach and the pancreas, and decreases the motility of both the stomach and the gut. Its physiological effects make it potentially useful in the therapy of various gastrointestinal diseases and disorders. Somatostatin is effective in the management of active variceal bleeding and increases the effectiveness of sclerotherapy. Previous studies show controversial data about the efficacy of somatostatin in preventing complications following endoscopic retrograde cholangiopancreatography. The use of this compound may be indicated only in high-risk patients in whom endoscopic sphincterotomy may also become necessary. The perioperative use of somatostatin decreases the risk of pancreatic surgery and its application is effective in promoting the closure of pancreatic fistulae. The somatostatin analogue octreotide has a definitive role in the medical management of hormone-secreting neuroendocrine tumours of the gut. PMID- 9867122 TI - Coeliac disease: always something to discover. AB - The authors present more than 20 years' experience with coeliac disease, with a summary of their published studies. Hair shaft characteristics were determined by scanning electron microscopy. Hair diameter was significantly lower and cuticular erosion scores higher in those who were not on gluten-free diets as compared to controls, showing a tendency towards normal values following start of gluten-free diets. Proton-induced X-ray emission showed significantly lower zinc content of the hair shaft in the group with acute coeliac disease and after a short-term diet, which approached the normal range only after a year-long diet. The serum prolactin levels in healthy controls and in coeliac patients on the diet were within normal limits, whereas in children with coeliac disease taking gluten in their meals, a significant hyperprolactinaemia was found. The erythrocyte glutathione content of coeliac children was elevated, and the glutathione disulfide level was significantly decreased, as compared to values in normal controls. The erythrocyte glutathione disulfide level and glutathione disulfide/erythrocyte glutathione ratio in coeliac children also differed from those in children with iron deficiency. With genotyping, the DQB1*0201/2 (p < 0.00001) and DR3 (p < 0.00001), DR7 (p < 0.01) alleles showed significant positive association with the disease. PMID- 9867123 TI - Neuropathy as an extrahepatic manifestation of chronic liver diseases. AB - Gastrointestinal motor disturbances and various cardiovascular symptoms are the characteristic features of autonomic neuropathy. Autonomic and sensory neuropathy has been described in chronic alcoholic liver diseases. Cardiovascular autonomic reflex tests as the gold standard for autonomic neuropathy and measuring of thresholds for constant current electric sine wave stimulation by neurometer were used for measuring sensory neuropathy in patients with chronic liver diseases of different etiology. Autonomic and sensory neuropathy was also observed in non alcoholic liver diseases. We proved that there is a correlation between autonomic neuropathy and prolongation of corrected QT interval in chronic liver diseases. We claim that autonomic neuropathy may have a role to play in the development of hyperdynamic circulation and portal hypertension in chronic liver diseases. PMID- 9867124 TI - Continued lack of access to the world's scientific community of the CTS Mouse. PMID- 9867125 TI - Differential cytokine mRNA expression in human labial minor salivary glands in primary Sjogren's syndrome. AB - Cytokines are known to be involved in a number of autoimmune conditions and are increasingly viewed as key components in numerous aspects of normal and abnormal cell functions. The purpose of the present study was to investigate possible immunopathogenic mechanisms within the labial minor salivary glands of patients with primary Sjogren's syndrome (pSS) by examining differential cytokine gene expression in individual cell populations (acini, ducts, or lymphoid cells). A cell-specific microdissection technique in combination with reverse transcription polymerase chain reaction (RT-PCR) and Southern hybridization using 32P-labeled cytokine gene-specific probes was utilized to measure cytokine messenger RNA expression in individual cell populations of patients and healthy controls. mRNAs for interleukin 2 (IL-2), interleukin 6 (IL-6), interleukin 10 (IL-10), tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma) and transforming growth factor beta 1 (TGF-beta1) were detected in the epithelial cells (acini and ducts) and lymphoid cells of the labial minor salivary glands of pSS patients. The expression levels of these mRNAs in the epithelial cells were either up- or down-regulated by adjacent focal infiltrating lymphoid cells. mRNAs for all of the above cytokines, with the exception of IFN-gamma, were detected in salivary tissues of healthy volunteers. The epithelial cells in the salivary glands are active participants in the autoimmune-mediated process of pSS, as evidenced by their ability to express a high frequency and wide variety of cytokines. The presence of an infiltrating lymphoid focus within the gland appeared to modulate cytokine gene expression by the salivary epithelial cells. PMID- 9867126 TI - Enhanced osteopontin expression and macrophage infiltration in MRL-Fas(lpr) mice with lupus nephritis. AB - MRL-Fas(lpr) mice spontaneously develop a chronic lupus-like renal disease, characterized by immune complex-mediated glomerulonephritis and abundant mononuclear cell infiltration in the interstitium. In the present study we have examined whether the macrophage chemoattractant osteopontin (Opn) could be important in the recruitment of macrophages in this murine model of autoimmune renal injury. We have examined the expression of Opn in the kidney of MRL Fas(lpr) mice and have correlated Opn synthesis with the degree of macrophage infiltration. Immunofluorescence staining revealed prominent expression of Opn by proximal tubules in MRL-Fas(lpr) mice but not in MRL-++ control mice. Northern blot analysis demonstrated that steady-state transcript levels for Opn mRNA were also significantly increased in MRL-Fas(lpr) kidneys compared with control kidneys. Furthermore, in situ hybridization showed massive Opn mRNA transcripts in proximal tubules in MRL-Fas(lpr) mice but not in controls. The diffuse macrophage infiltration in the kidney of MRL-Fas(lpr) correlated with the enhanced Opn expression. Opn secretion in vitro by cultured renal tubular epithelial cells was upregulated by TNF-alpha and 1,25(OH)2-vitamin D3, whereas no regulation was observed in a control macrophage cell line. We conclude that the enhanced expression of the chemotactic molecule Opn by tubular cells is a prominent feature of murine lupus nephritis and might be promoted by the proinflammatory cytokine environment in MRL-Fas(lpr). The chronic upregulation of Opn could participate in the recruitment of monocytes in the kidney of MRL Fas(lpr) mice, thereby contributing to the pathogenesis of autoimmune renal disease. PMID- 9867127 TI - Spontaneous occurrence of anti-fibrillin-1 autoantibodies in tight-skin mice. AB - Tight-skin (TSK) mouse represents an experimental for systemic sclerosis, displaying cutaneous hyperplasia, connective tissue alterations in the internal organs and developing autoantibodies against several scleroderma target autoantigens. TSK mouse syndrome is associated with a mutation in fibrillin-1 (Fbn-1), the major component of 10 nm microfibrils. Here, we have investigated whether TSK mouse develops autoimmunity to Fbn-1 similar to scleroderma target autoantigens. Our results show that anti-Fbn-1 IgG autoantibodies are present in high titer in many TSK mice. Specificity of these antibodies was confirmed by competitive inhibition assays and Western blotting analysis using recombinant human Fbn-1 protein. TSK mouse autoantibodies recognize a conserved epitope present in the C region of Fbn-1. These results indicate the presence of Fbn-1 specific T and B cells in TSK mouse repertoire. PMID- 9867128 TI - Dapsone at onset of diabetes lowers glycated hemoglobin and delays death in NOD mice. AB - Dapsone (4,4'-diaminodiphenyl sulfone) is a compound that has a large clinical experience due to its antimicrobial effects against mycobacterium leprae, the causative agent of leprosy. It is increasingly used in a number of clinical situations where inflammation may play an ancillary role. An inhibitory effect of the drug or lack thereof in the cumulative incidence or propagation of diabetes mellitus in the NOD mouse has mechanistic as well as therapeutic implications. We previously showed that dapsone administered continuously as a percentage of food to NOD mice inhibits the cumulative incidence of diabetes in a dose dependent fashion. In the present experiment, female NOD litter mates were randomized to receive dapsone (0.001% w/w as a percentage of food) at onset of diabetes. There were no differences in weight, blood glucose, or glycated hemoglobin at 10 weeks of age among the animals that were ultimately to receive dapsone (n = 10), mouse chow alone (n = 9), or those who did not develop diabetes (n = 3). The mean time to onset of diabetes, mean blood glucose at onset, and mean glycated hemoglobin at onset did not differ between animals who did or did not receive dapsone. Animals receiving dapsone had significantly (p < or = 0.03) lower glycated hemoglobin at weeks 2, 3, and 4 following the onset of diabetes and lived significantly longer following diagnosis of diabetes (7 vs. 4 weeks, p < 0.05). In conclusion, dapsone modulates the progression of autoimmune diabetes in the NOD mouse even when administered after the initiation of hyperglycemia. PMID- 9867129 TI - Semi-specific immuno-absorption and monoclonal antibody therapy in ANCA positive vasculitis: experience in four cases. AB - The treatment of renal limited systemic vasculitis usually involves a combination of cytotoxic drugs and steroids. As shown by randomised prospective controlled trial, plasmapheresis may be of additional benefit for the management of patients with renal involvement severe enough to require dialysis support. Recently, growing evidence has suggested that autoantibodies to neutrophil cytoplasm (ANCA) may play a role in the pathogenesis of the primary vasculitides by promoting neutrophil mediated endothelial cell cytotoxicity. This has led to new strategies for treatment based on firstly, the use of semi-specific immunoabsorption (IA) devices to remove circulating autoantibodies, and secondly, the use of 'Humanised' monoclonal antibodies (MAbs) with specificity for lymphocytes, particularly T lymphocytes. We have treated four patients, two with ANCA specificity for proteinase 3 (PR3), and two with specificity for myeloperoxidase (MPO). Semi-specific IA was carried out by plasmapheresis through extracorporeal online devices, using L tryptophan as the immobilised immunoabsorbant. Of the four patients who received IA, three showed substantial depletion in ANCA titres and resolution of clinical symptoms. The MAbs were subsequently used to attempt to obtain long-term control of ANCA synthesis. These results suggest that an optimal strategy for treatment of systemic vasculitis might consist of specific IA, using immobilised ANCA antigens to deplete circulating vasculotoxic antibodies, combined with MAb therapy to restore immune homeostasis. PMID- 9867130 TI - T lymphocyte activation in myasthenic thymoma. AB - The functional and phenotypic characteristics of lymphocytes separated from myasthenic thymoma (Th-L) were compared with those of lymphocytes separated from non-thymomatous thymus associated with thymoma (NTh-L) of the same patients and NTh-L of myasthenia gravis (MG) patients without thymoma. We examined whether Th L and NTh-L of MG patients reacted to interleukin-2 (IL-2) to develop lymphokine activated killer (LAK) activity and/or cytolytic activity against K562 (natural killer (NK) activity), and the phenotypic changes in such cells during incubation. Ten MG patients with thymoma and six MG patients without thymoma, and four non-MG thymoma patients were included in this study. Th-L and NTh-L of MG patients reacted with IL-2 to develop LAK and NK activities. The LAK activity developed from Th-L was significantly higher than that from NTh-L, and the LAK activity developed from Th-L was as high as that from peripheral blood lymphocytes (PBL) in MG patients without thymoma. The proportions of CD3+ cells, CD4+/CD8- cells, and CD4-/CD8+ cells in Th-L of MG patients increased significantly during incubation. On the other hand, the proportion of CD4+/CD8+ cells in Th-L of MG patients decreased significantly. The proportions of CD4+/CD8 cells, CD4-/CD8+ cells, and CD4+/CD8+ cells in NTh-L of MG patients with and without thymoma exhibited no change during incubation. These findings suggest that CD4+/CD8+ Th-L of MG patients may have a higher potential to react to IL-2 than NTh-L, and that the former cells might develop LAK activity like that of PBL on maturation to CD4+/CD8- cells and CD4-/CD8+ cells. Our findings also suggested that Th-L might play an important role in the pathogenesis of MG with thymoma. PMID- 9867131 TI - Advancing medical care: the role of nuclear medicine in radioguided surgery, part II: sentinel node dissections. PMID- 9867132 TI - The role of lymphoscintigraphy in lymphatic mapping for melanoma and breast cancer. PMID- 9867133 TI - How technetium was discovered in a pile of junk. PMID- 9867134 TI - Fasting and nonfasting iodine-123-idophenylpentadecanoic acid myocardial SPECT imaging in coronary artery disease. AB - Iodine-123-labeled idophenylpentadecanoic acid (IPPA) metabolic imaging has been shown to be clinically useful for the identification of myocardial viability in patients with coronary artery disease and left ventricular dysfunction. Imaging is usually performed under fasting conditions since nonfasting conditions may affect myocardial uptake of 123I-IPPA. The purpose of this study was to examine the impact of dietary condition on 123I-IPPA metabolic imaging. METHODS: Forty patients with stable coronary artery disease underwent, in randomized order and on separate days, 123I-IPPA SPECT myocardial imaging under fasting and nonfasting conditions. Patients were injected with 123I-IPPA (4-5 mCi) at rest with imaging performed at 4 (initial) and 30 (delay) min. For each image (initial and delay images), 10 segments were analyzed by three experienced observers without knowledge of patient identity or dietary condition using a 5-point grading system (O = no uptake to 4 = normal uptake). A summed global score was obtained for each image by adding the scores for all 10 segments. Image quality was assessed using a 3-point grading system. RESULTS: Visual agreement for normal and abnormal segments between fasting and nonfasting conditions was 82% (kappa = 0.63). There were no significant differences in the summed global scores for both conditions. Image quality was equivalent for both conditions in 65% of cases and superior under the nonfasting condition in 25% of cases. CONCLUSION: Image quality as well as the presence, location and severity of defects are similar under fasting and nonfasting conditions with 123I-IPPA. Therefore, fasting is not necessary before 123I-IPPA SPECT imaging for the assessment of myocardial viability. PMID- 9867135 TI - High-resolution cardiac PET in rabbits: imaging and quantitation of myocardial blood flow. AB - A high-resolution PET system for small animals was tested for its applicability to the investigation of regional myocardial blood flow (MBF) in rabbits. METHODS: Nineteen measurements were performed in 10 closed-chest anesthetized rabbits at baseline and during infusions of adenosine (0.2 mg/kg/min) and propranolol (0.20 1.20 mg slow infusion) to obtain a wide range of MBF. Myocardial blood flow was assessed both by dynamic 13N-ammonia PET and by colored microspheres. Blood was withdrawn directly from the femoral artery, and arterial 13N activity was measured by coincidence type gamma detection system for the input function. Nitrogen-13 myocardial uptake was calculated by dividing the myocardial 13N activity by the integral value of the input function. RESULTS: Three or four contiguous cross-sectional myocardial images were obtained after 13N-ammonia injection. The left ventricular wall and cardiac cavity were clearly visualized. Moreover, initial passage of the tracer through the heart was obtained with serial 10-sec PET images. Nitrogen-13 myocardial uptake correlated well with flow measured with microspheres (r = 0.88). CONCLUSION: Our cardiac PET system can be used for in vivo imaging and quantitation of MBF in small animals and may play an important role in the future study of animal models of cardiovascular diseases. PMID- 9867136 TI - Comparison of four motion correction techniques in SPECT imaging of the heart: a cardiac phantom study. AB - The aim of this study was to evaluate the accuracy of four different motion correction techniques in SPECT imaging of the heart. METHODS: We evaluated three automated techniques: the cross-correlation (CC) method, diverging squares (DS) method and two-dimensional fit method and one manual shift technique (MS) using a cardiac phantom. The phantom was filled with organ concentrations of 99mTc closely matching those seen in patient studies. The phantom was placed on a small sliding platform connected to a computer-controlled stepping motor. Linear, random, sinusoidal and bounce motions of magnitude up to 2 cm in the axial direction were simulated. Both single- and dual-detector 90 degrees acquisitions were acquired using a dual 90 degrees detector system. Data were acquired over 180 degrees with 30 or 15 frames/detector (single-/dual-head) at 30 sec/frame in a 64x64 matrix. RESULTS: The simulated single-detector system, CC method, failed to accurately correct for any of the simulated motions. The DS technique overestimated the magnitude of phantom motion, particularly for images acquired between 45 degrees left anterior oblique and 45 degrees left posterior oblique. The two-dimensional and MS techniques accurately corrected for motion. The simulated dual 90 degrees detector system, CC method, only partially tracked random or bounce cardiac motion and failed to detect sinusoidal motion. The DS technique overestimated motion in the latter half of the study. Both the two dimensional and MS techniques provided superior tracking, although no technique was able to accurately track the rapid changes in cardiac location simulated in the random motion study. Average absolute differences between true and calculated position of the heart on single- and dual 90 degrees -detectors were 1.7 mm and 1.5 mm for the two-dimensional and MS techniques, respectively. The corresponding values for the DS and CC techniques were 5.7 and 8.9 mm, respectively. CONCLUSION: Of the four techniques evaluated, manual correction by an experienced technologist proved to be the most accurate, although results were not significantly different from those observed with the two-dimensional method. Both techniques accurately determined cardiac location and permitted artifact-free reconstruction of the simulated cardiac studies. PMID- 9867137 TI - Technetium-99m-methylene disphosphonate (MDP) bone scan hypercalcemia. PMID- 9867138 TI - Fluorine-18-fluorodeoxyglucose cardiac imaging using a modified scintillation camera. AB - Conventional 201TI and hexakis 2-methoxy-2-isobutyl isonitrile studies are less accurate as compared to FDG PET in the prediction of functional recovery after revascularization in patients with injured but viable myocardium. The introduction of a dual-head variable-angle-geometry scintillation camera equipped with thicker crystals (5/8 in.) and high-resolution, ultrahigh-energy collimators capable of 511 keV imaging has permitted FDG SPECT to provide information equivalent to that of PET for the detection of injured but viable myocardium in patients with chronic ischemic heart disease. The development of standardized glucose-loading protocols, including glucose-insulin-potassium infusion and the potential use of nicotinic acid derivatives, has simplified the method of obtaining consistently good-to-excellent quality FDG SPECT cardiac studies. FDG SPECT may become the modality of choice for evaluating injured but viable myocardium because of enhanced availability of FDG, logistics, patient convenience, accuracy and cost-effectiveness compared to PET. PMID- 9867139 TI - Crossed cerebellar diaschisis due to intracranial hematoma in basal ganglia or thalamus. AB - The purpose of our study was to evaluate the remote effects on the cerebellum and cerebral cortex from subcortical hematoma without cortical structural abnormality. METHODS: Our study included 23 patients with hematoma, strictly confined either to the basal ganglia (n = 12) or thalamus (n = 11) without cortical abnormality on CT or MRI. Twenty psychiatric patients without structural abnormality on MRI were selected as control subjects. Technetium-ethyl cysteinate dimer brain SPECT was performed in patients and control subjects. Regional cerebral blood flow (rCBF) was visually and semiquantitatively assessed. Asymmetry index (AI) was determined using data from regions of interest at the basal ganglia, thalamus, cerebellum, frontal, parietal and temporal cortex to support the semiquantitative analysis. The criteria for defining hypoperfusion that reflected diaschisis was based on an AI > the mean + 2 s.d. of AI in control subjects. RESULTS: In the basal ganglia hematoma, rCBF was reduced significantly in the contralateral cerebellum (10/12), ipsilateral thalamus (12/12), ipsilateral frontal (6/12), parietal (12/12) and temporal cortex (10/12). As for thalamic hematoma, significantly reduced perfusion was seen in the contralateral cerebellum (10/11), ipsilateral basal ganglia (7/11), ipsilateral frontal (5/11), parietal (11/ 11) and temporal cortex (3/11). CONCLUSION: Crossed cerebellar diaschisis (CCD) and cortical diaschisis frequently were observed in patients with subcortical hematoma without cortical structural abnormality. This suggested that CCD can develop regardless of interruption of the corticopontocerebellar tract, which is the principal pathway of CCD. PMID- 9867140 TI - PET quantification of specific binding of carbon-11-nicotine in human brain. AB - Previous work on the PET measured uptake of (S)-[11C]nicotine presents conflicting findings as to whether it reflects specific binding. METHODS: We studied the uptake of (R)-[11C]nicotine and (S)-[11C]nicotine in normal volunteers at baseline conditions and after a challenge with unlabeled (S) nicotine to decrease the concentration of free binding sites or with CO2 to increase perfusion. We analyzed the data using two- and three-compartment models. RESULTS: We found tissue pharmacokinetics of (R)- and (S)-[11C]nicotine are adequately described by the two-compartment model. (S)-nicotine challenge induced small but statistically significant reductions in distribution volume (DV) of both (R)- and (S)-[11C]nicotine. The changes in DV could not be attributed to perfusion changes because DV was not affected by CO2 challenge. Although the reduction in DV indicates sensitivity of [11C]nicotine to status of nicotinic binding sites, the small magnitude of the reduction suggests that most nicotine uptake is nonspecific. CONCLUSION: Although differences in DV attributable to specific binding were detected, (R)- and (S)-[11C]nicotine are relatively poor tracers for studying nicotinic binding sites using PET. PMID- 9867141 TI - Regional cerebral blood flow scintigraphy in tick-borne encephalitis and other aseptic meningoencephalitis. AB - In a prospective study, regional cerebral blood flow (rCBF) was studied in patients with aseptic meningoencephalitis at 6 wk and 1 yr after onset of disease. METHODS: Patients with tick-borne encephalitis ([TBE] n = 73) and meningoencephalitis of other etiology ([non-TBE] n = 56) were investigated with rCBF-scintigraphy (SPECT). SPECT images in the acute phase of disease and at long term follow-up were analyzed for blood-flow disturbances and their localization in the central nervous system and were correlated to clinical course and outcome. RESULTS: Decreased rCBF was seen in 50% of patients after 6 wk (TBE 49%, non-TBE 50%) and in 46% (TBE 47%, non-TBE 46%) after 1 yr. The decrease in rCBF was moderate in 18% and 11% at 6 wk and in 8% and 9% at the 1-yr follow-up of TBE and non-TBE patients, respectively. Reduced rCBF was significantly more common among patients with encephalitis than among those with meningitis, and more common in males. The distribution of cerebral flow changes was predominantly patchy or multifocal. At long-term follow-up, improvement in rCBF was seen in 28 of 109 patients (26%), but worsening of decreased rCBF was demonstrated in 19 of 109 (17%). In TBE patients, remaining neurological symptoms at 6 wk of disease were associated with worsening of decreased rCBF at the 1-yr follow-up. CONCLUSION: With SPECT, rCBF changes, mostly slight and patchy or multifocal, were detected in patients with aseptic meningoencephalitis. Decreased rCBF was more frequent in patients with moderate-to-severe encephalitis, although the clinical use in predicting long-term outcomes in aseptic meningoencephalitis (e.g., TBE) seems limited. PMID- 9867142 TI - Carbon-11-NNC 112: a radioligand for PET examination of striatal and neocortical D1-dopamine receptors. AB - The aim of this work was to explore the potential of a selective D1-dopamine receptor antagonist as a new radioligand for PET examination of striatal and neocortical D1-dopamine receptors. METHODS: The active (+)- and inactive (-) enantiomers of [11C]NNC 112 were radiolabeled using the N-methylation approach and were examined by PET in cynomolgus monkeys and healthy men. Metabolite levels in plasma were measured by gradient high-performance liquid chromatography (HPLC). RESULTS: N-methylation of the corresponding desmethyl precursors with [11C]methyl triflate gave high total radiochemical yield (50%-60%) and specific radioactivity (110 GBq/micromol). (+)-[11C]NNC 112 binding in cynomolgus monkeys was 5.77+/-0.31 and 2.36+/-0.14 times higher in the striatum and neocortex, respectively, than in the cerebellum at a transient equilibrium that appeared 40 50 min after injection. The binding of (+)-[11C]NNC 112 is stereoselective, because the brain distribution of the inactive (-)-enantiomer was on an equally low level for all brain regions. Displacement and pretreatment experiments using unlabeled SCH 23390 and ketanserin confirms that (+)-[11C]NNC 112 binds specifically and reversibly to D1-dopamine receptors. The radioactivity ratios of the striatum, frontal cortex and nucleus accumbens to the cerebellum were 3.8 4.0, 1.7-2.0 and 2.8-3.1, respectively, at a transient equilibrium that appeared 40-50 min after injection in four healthy human subjects. Linear graphical analysis gave distribution volume ratios of 3.9 and 1.5 in the putamen and frontal cortex, respectively. The fraction of the total radioactivity in human plasma representing unchanged (+)-[11C]NNC 112 was 85% at 5 min and 25% at 75 min after injection. CONCLUSION: (+)-[11C]NNC 112 should be a useful PET radioligand for quantitative examination of not only striatal but neocortical D1-dopamine receptors in man. PMID- 9867143 TI - Biodistribution and dosimetry of TRODAT-1: a technetium-99m tropane for imaging dopamine transporters. AB - Technetium-99m TRODAT-1 is an analog of cocaine that selectively binds the presynaptic dopamine transporters. The primary purpose of this study was to measure its whole-body biokinetics and radiation dosimetry in healthy human volunteers. The study was conducted within a regulatory framework that required its pharmacological safety to be assessed simultaneously. METHODS: The sample included 4 men and 6 women ranging in age from 22-54 yr. An average of 20 whole body scans were acquired sequentially on a dual-head camera for up to 46 hr after the intravenous administration of 370+/-16 MBq (10.0+/-0.42 mCi) 99mTc TRODAT. The renal excretion fractions were measured from 12-24 discrete urine specimens. The fraction of the administered dose in 17 regions of interest and each urine specimen was quantified from the attenuation and background corrected geometric mean counts in conjugate views. Multiexponential functions were iteratively fit to each time-activity curve using a nonlinear, least squares regression algorithm. These curves were numerically integrated to yield source organ residence times. Gender-specific radiation doses were then estimated with the Medical Internal Radiation Dose technique for each subject individually before any results were averaged. RESULTS: There were no pharmacological effects of the radiotracer on any of the subjects. The early planar images showed differentially increased activity in the nose, pudendum and stomach. SPECT images demonstrated that the radiopharmaceutical localized in the basal ganglia in a distribution that was consistent with selective transporter binding. Image analysis showed that the kidneys excreted between 20% and 32% of the injected dose during the first 22-28 hr postadministration, after which no more activity could be recovered in the urine. The dose limiting organ in both men and women was the liver, which received an average of 0.046 mGy/MBq (0.17 rads/mCi, range 0.14-0.22 rad/mCi). In the worst case, which was clearly an over-estimation, it would have taken 22.7 mCi to deliver 5 rad to the liver. CONCLUSION: TRODAT may be a safe and effective radiotracer for imaging dopamine transporters in the brain and the body. PMID- 9867144 TI - Radiotoxicity after strontium-89 therapy for bone metastases using the micronucleus assay. AB - The purpose of this study was to evaluate the degree of cytologic radiation damage to lymphocytes after 89Sr therapy using the cytokinesis-blocked micronucleus assay. The chromosomal damage to lymphocytes exposed to 89Sr in vivo should result in augmentation of the number of cells with micronucleus. METHODS: We studied eight patients with painful bone metastases, who were treated with 111 MBq89Sr. Isolated lymphocytes collected from the patients 1 wk after therapy were harvested and treated according to the cytokinesis-blocked method of Fenech and Morley. The number of micronuclei per 500 binucleated cells was scored by visual inspection. As controls, lymphocytes from the same patients before therapy were also studied. For three patients, serial blood samples were examined for a maximum of 2 mo after therapy. In an in vitro study, lymphocytes from five normal volunteers were exposed to doses varying from 0.25 to 1.0 Gy and studied with the same method. RESULTS: The mean number (+/-s.d.) of micronuclei per 500 binucleated cells after treatment was significantly increased (p<0.05) as compared to control subjects (17.1+/-3.0 compared to 6.0+/-1.7). Thereafter, the number of micronuclei recovered gradually by 6 wk following therapy and, in one case, nearly to the baseline range in 2 mo. The number of micronuclei after 0.53+/-0.13 Gy of external irradiation was nearly equivalent to that after 89Sr therapy. CONCLUSION: The relatively low frequency of lymphocyte micronuclei exposed to 89Sr in vivo supported the contention that short-term nonstochastic damage with 111 MBq89Sr in patients with painful bone metastases is minimal. PMID- 9867145 TI - Lymphoscintigraphy and radioguided biopsy of the sentinel axillary node in breast cancer. AB - Lymphoscintigraphy associated with radioguided biopsy of the sentinel node (SN) is well established in clinical practice for melanoma. In breast cancer, the SN concept is similarly valid, and lymphoscintigraphy is a useful method for localizing the axillary SN. The aim of this study was to optimize the lymphoscintigraphy technique in association with a gamma ray detecting probe (GDP) for identifying and removing the SN in breast cancer patients. METHODS: Two hundred fifty patients with operable breast tumor underwent lymphoscintigraphy before surgery. Three different size ranges of 99mTc-labeled colloid particles (<50, <80 and 200-1000 nm) were used, with either subdermal (above tumor) or peritumoral injection. Early and late scintigraphic images were obtained in anterior and oblique projections, and the skin projection of the detected SN was marked. Sentinel nodes were identified and removed with the aid of the GDP during breast surgery; they were tagged separately. Complete axillary dissection followed. In 40 patients, a blue dye was also administered in addition to subdermal radiolabeled colloid to compare blue dye mapping with lymphoscintigraphy localization. RESULTS: Lymphoscintigraphy successfully revealed lymphatic drainage in 245 of 250 patients (98%). The axillary SN was identified in 240 patients (96%). SN biopsy correctly predicted axillary node status in 234 of 240 patients (97.5%). Lymphoscintigraphy and GDP detected the SN most easily and consistently when 200-1000 nm colloid was administered subdermally in an injection volume of 0.4 ml. Blue dye mapping was successful in 30 of 40 patients (75%). In 26 of these patients, the dye and lymphoscintigraphy identified the same node; in 4 cases different nodes were identified. None of these four patients had axillary disease. CONCLUSION: Lymphoscintigraphy is a simple procedure that is well tolerated by patients. Sentinel node identification is more reliable when large-size radiolabeled colloids are injected in a relatively small injection volume (0.4 ml). Use of a GDP greatly facilitates precise pinpointing and rapid removal of the SN. PMID- 9867146 TI - Fluorine-18-fluorodeoxyglucose PET identification of cardiac metastasis arising from uterine cervical carcinoma. AB - Cardiac metastasis of uterine cervical carcinoma is rare. We describe a patient with a past history of uterine cervical carcinoma who presented with metastasis to the heart, lungs and distant lymph nodes 3 yr after surgery and chemotherapy. Since the patient complained of chest pain and demonstrated electrocardiogram abnormalities, we performed echocardiography, electron beam CT and MRI, which revealed a tumor in the right ventricular wall. The tumor was assessed by 67Ga scintigraphy and 18F-fluorodeoxyglucose (FDG) PET scanning. The mean differential 18F-FDG uptake ratio of the tumor was 7.9, suggesting malignancy, which was later confirmed by myocardial biopsy. Information about the extent of the tumor and partial necrosis within it was provided by 18F-FDG PET. Although both radionuclide imaging techniques also detected metastatic lesions in the lungs and lymph nodes, 18F-FDG PET scanning detected small lesions more sensitively than 67Ga scintigraphy. PMID- 9867147 TI - PET evaluation of therapeutic limb perfusion in Merkel's cell carcinoma. AB - An 87-yr-old woman diagnosed with recurrent Merkel's cell carcinoma was treated with therapeutic limb perfusion and underwent PET scanning with 18F fluorodeoxyglucose (FDG). PET studies were obtained before and after treatment to determine the response to the intervention. A baseline whole-body study was obtained to assess the extent and degree of disease activity. This was followed by a repeat PET scan 2 mo. later after treatment with isolated limb chemotherapy with high-dose melphalan and tumor necrosis factor-alpha. The initial scan demonstrated multiple foci of high FDG uptake in the left calf, a left supraclavicular lesion and also detected concurrent keratinizing squamous cell metastasis in the right axilla. A repeat PET study showed complete metabolic resolution of the lesions in the left calf after treatment. FDG PET may be a useful technique for staging Merkel cell carcinoma and for assessing the tumor response after therapy of this rare tumor. PMID- 9867148 TI - Response to treatment with yttrium 90-DOTA-lanreotide of a patient with metastatic gastrinoma. AB - 1,4,7,10-tetraazacyclododecane-N,N',N",N'''-tetraacetic acid (DOTA)-lanreotide is a universal somatostatin (SST) receptor subtype ligand that binds to a large variety of human tumors. We report the case of a patient with metastatic gastrinoma who was treated with 90Y-DOTA-lanreotide. Before treatment, dosimetry with 111In-DOTA-lanreotide (150 MBq, 10 nmol) indicated a dose of 5.8 mGy/MBq for the recurrent abdominal gastrinoma, and a mean dose of approximately 1.0 mGy/MBq for liver metastases (i.e., 56 and approximately 10 mGy/MBq for 90Y-DOTA lanreotide, respectively). After four infusions of 90Y-DOTA-lanreotide (each 1 GBq, approximately 30 nmol) over a 6-mo period, the 111In-DOTA-lanreotide scintigraphy of the liver had returned to a nearly normal condition and a remarkably decreased uptake by the recurrent gastrinoma was calculated (approximately 5 mGy/MBq for 90Y-DOTA-lanreotide). The imaging results were well correlated with a 25% regression of the liver metastases as indicated by CT. Blood, urine and whole-body clearances of 111In-DOTA-lanreotide and 90Y-DOTA lanreotide were very similar. The DOTA-lanreotide promises to be useful for functional tumor diagnosis (111In-DOTA-lanreotide) and receptor-mediated tumor radiotherapy (90Y-DOTA-lanreotide). PMID- 9867149 TI - Flare response seen in therapy for osteomalacia. AB - We report an interesting case of osteomalacia in which flare response was seen during therapy. The first 99mTc-methylene diphosphonate bone scan showed increased bilateral and symmetric uptake in the ribs, clavicles and iliac bones. Thoracic CT showed symmetric radiolucent seams (Looser's zones) in both ribs, which were pathognomonic of osteomalacia. After initiation of therapy with vitamin D, the patient's subjective symptoms gradually were relieved. On a second bone scan 4 mo. after initiation of therapy, the hot spots in the ribs remained unchanged. Uptake in the bilateral clavicles had become more intense, and new hot spots were recognized in the right lower ribs and left tibia. A third bone scan after 10 mo. demonstrated an obvious decrease in the number and intensity of the hot spots. Increased uptakes in the second scan were thought to be a flare response caused by therapy. PMID- 9867150 TI - Dose escalation trial of indium-111-labeled anti-carcinoembryonic antigen chimeric monoclonal antibody (chimeric T84.66) in presurgical colorectal cancer patients. AB - Chimeric T84.66 (cT84.66) is a high-affinity (1.16x10(11) M(-1)) IgG1 monoclonal antibody against carcinoembryonic antigen (CEA). The purpose of this pilot trial was to evaluate the tumor-targeting properties, biodistribution, pharmacokinetics and immunogenicity of 111In-labeled cT84.66 as a function of administered antibody protein dose. METHODS: Patients with CEA-producing colorectal cancers with localized disease or limited metastatic disease who were scheduled to undergo definitive surgical resection were each administered a single intravenous dose of 5 mg of isothiocyanatobenzyl diethylenetriaminepentaacetic acid-cT84.66, labeled with 5 mCi of 111In. Before receiving the radiolabeled antibody, patients received unlabeled diethylenetriaminepentaacetic acid-cT84.66. The amount of unlabeled antibody was 0, 20 or 100 mg, with five patients at each level. Serial blood samples, 24-hr urine collections and nuclear images were collected until 7 days postinfusion. Human antichimeric antibody response was assessed up to 6 mo postinfusion. RESULTS: Imaging of at least one known tumor site was performed in all 15 patients. Fifty-two lesions were analyzed, with an imaging sensitivity rate of 50.0% and a positive predictive value of 76.9%. The antibody detected tumors that were not detected by conventional means in three patients, resulting in a modification of surgical management. Interpatient variations in serum clearance rates were observed and were secondary to differences in clearance and metabolic rates of antibody and antibody:antigen complexes by the liver. Antibody uptake in primary tumors, metastatic sites and regional metastatic lymph nodes ranged from 0.4% to 134% injected dose/kg, resulting in estimated 90Y-cT84.66 radiation doses ranging from 0.3 to 193 cGy/mCi. Thirteen patients were evaluated 1-6 mo after infusion for human antichimeric antibody, and none developed a response. No major differences in tumor imaging, tumor uptake, pharmacokinetics or organ biodistribution were observed with increasing protein doses, although a trend toward increasing blood uptake and decreasing liver uptake was observed with increasing protein dose. CONCLUSION: Chimeric T84.66 demonstrated tumor targeting comparable to other radiolabeled intact anti-CEA monoclonal antibodies. Its immunogenicity after single administration was lower than murine monoclonal antibodies. These properties make 111In-cT84.66, or a lower molecular weight derivative, attractive for further evaluation as an imaging agent. Yttrium-90 dosimetry estimates predict potentially cytotoxic radiation doses to select tumor sites, which makes 90Y-cT84.66 also appropriate for further evaluation in Phase I radioimmunotherapy trials. Although clinically important changes in biodistribution, pharmacokinetics and tumor targeting with increasing protein doses of 111In-cT84.66 were not demonstrated, the results do suggest that antibody clearance from the blood is driven by hepatic uptake and metabolism, with more rapid blood clearance seen in patients with liver metastases. These patients with rapid clearance and potentially unfavorable biodistribution for imaging and therapy may, therefore, be a more appropriate subset in which to evaluate the role of administering higher protein doses. This underscores the need to further identify, characterize and understand those factors that influence the biodistribution and clearance of radiolabeled anti-CEA antibodies, to allow for better selection of patients for therapy and rational planning of radioimmunotherapy. PMID- 9867151 TI - Optimized conditions for chelation of yttrium-90-DOTA immunoconjugates. AB - Radioimmunotherapy (RIT) with 90Y-labeled immunoconjugates has shown promise in clinical trials. The macrocyclic chelating agent 1,4,7,10-tetraazacyclododecane N,N',N",N"'-tetraacetic acid (DOTA) binds 90Y with extraordinary stability, minimizing the toxicity of 90Y-DOTA immunoconjugates arising from loss of 90Y to bone. However, reported 90Y-DOTA immunoconjugate product yields have been typically only < or =50%. Improved yields are needed for RIT with 90Y-DOTA immunoconjugates to be practical. METHODS: (S) 2-[p-(bromoacetamido)benzyl]-DOTA (BAD) was conjugated to the monoclonal antibody Lym-1 via 2-iminothiolane (2IT). The immunoconjugate product, 2IT-BAD-Lym-1, was labeled in excess yttrium in various buffers over a range of concentrations and pH. Kinetic studies were performed in selected buffers to estimate radiolabeling reaction times under prospective radiopharmacy labeling conditions. The effect of temperature on reaction kinetics was examined. Optimal radiolabeling conditions were identified and used in eight radiolabeling experiments with 2IT-BAD-Lym-1 and a second immunoconjugate, DOTA-peptide-chimeric L6, with 248-492 MBq (6.7-13.3 mCi) of 90Y. RESULTS: Ammonium acetate buffer (0.5 M) was associated with the highest uptake of yttrium. On the basis of kinetic data, the time required to chelate 94% of 90Y (four half-times) under prospective radiopharmacy labeling conditions in 0.5 M ammonium acetate was 17-148 min at pH 6.5, but it was only 1-10 min at pH 7.5. Raising the reaction temperature from 25 degrees C to 37 degrees C markedly increased the chelation rate. Optimal radiolabeling conditions were identified as: 30-min reaction time, 0.5 M ammonium acetate buffer, pH 7-7.5 and 37 degrees C. In eight labeling experiments under optimal conditions, a mean product yield (+/- s.d.) of 91%+/-8% was achieved, comparable to iodination yields. The specific activity of final products was 74-130 MBq (2.0-3.5 mCi) of 90Y per mg of monoclonal antibody. The immunoreactivity of 90Y-labeled immunoconjugates was 100%+/-11%. CONCLUSION: The optimization of 90Y-DOTA chelation conditions represents an important advance in 90Y RIT because it facilitates the dependable and cost-effective preparation of 90Y-DOTA pharmaceuticals. PMID- 9867152 TI - Quantifying the radiation dosage to individual skeletal lesions treated with samarium-153-EDTMP. AB - Samarium-153ethylenediaminetetramethylenephosphonate (EDTMP) is used in the treatment of painful skeletal lesions. This study attempted to quantify the radiation dosage to individual lesions on both the macroscopic and microscopic level. METHODS: A gamma camera-based quantification technique was adapted and refined for 153Sm. The accuracy of the technique was determined by using a realistic phantom. The activity and volume of lesions as well as normal bone were determined and used to estimate the radiation dosages to these regions. Two patients died of unrelated causes shortly after receiving 153Sm-EDTMP. This made it possible to compare the gamma camera results with direct measurements. It also allowed for autoradiographic examination of the lesions. Finally, the microscopic radiation dosages were estimated. RESULTS: The phantom study indicated that the quantification technique was off, on average, by 4.1% (s.d. = 8.1%). The absolute activity concentration of trabecular bone was found to be approximately 0.22 MBq/g, and that of cortical bone was found to be approximately 0.1 MBq/g, regardless of the dosage administered. The corresponding concentrations for lesions were between 3 and 7 times higher than that of normal bone, with no apparent ceiling. From these results, the macroscopic radiation dosage could be estimated. The dosage to normal bone varied between 0.9 and 3.9 cGy x kg/MBq, and that of the lesions varied between 5.2 and 27.1 cGy x kg/MBq. The autopsy results confirmed that the gamma camera technique was accurate. The autoradiography showed clearly that the activity was associated with the surface of the bone. From these findings, the microscopic radiation dosage distribution was estimated for cortical and trabecular bone as well as osteoblastic lesions. The variation in the microscopic dosage compared to the macroscopic dosage was quite large. Microscopic dosages, when compared to the macroscopic dosages, were as high as 965% and as low as 14.9%. CONCLUSION: The techniques used have been proven to be accurate. The activity in normal bone may be at a ceiling value for all the administered doses, which could explain the small variation. This is not true for the lesions. The large variation in dosages on a microscopic scale, combined with the ceiling in normal bone, may explain the lower than expected toxicity and relatively quick relapse of the patients. PMID- 9867153 TI - Technetium-99m-tetrofosmin scintigraphy in pulmonary tuberculosis. AB - Technetium-99m-tetrofosmin, an agent that is widely used in myocardial imaging, has been reported to accumulate in several types of malignancies, including lung tumors. Yet, there is limited knowledge about its role in imaging infection or inflammatory lesions. The aim of this study was to investigate the role of 99mTc tetrofosmin scintigraphy in pulmonary tuberculosis in cases with active and inactive tuberculosis in comparison with radiological and microbiological findings. METHODS: Twenty-seven patients with active pulmonary tuberculosis (APTB) and 6 patients with inactive pulmonary tuberculosis (IPTB), proven by sputum smears and cultures, were included in this study. Mean age of the group was 42.6+/-13 yr. Nine months after therapy, 99mTc-tetrofosmin scintigraphy was repeated in 6 patients with APTB to evaluate response to therapy. Ten-minute anterior and posterior chest images were acquired 20 and 60 min after the injection of 370 MBq (10 mCi) 99mTc-tetrofosmin. The images were evaluated both visually and semiquantitatively by two blinded nuclear medicine physicians. For semiquantitative evaluation, regions of interest (ROIs) were drawn over the lesion (L) and nonlesion areas (NL). The mean count values of ROIs were obtained and L/NL ratios were calculated. RESULTS: According to the visual evaluations, 99mTc-tetrofosmin uptake was Grade (+) in 4 (15%) and Grade (++) in 23 (85%) patients with APTB. Technetium-99m-tetrofosmin uptake was negative in 5 patients with IPTB. Grade (+) 99mTc-tetrofosmin uptake was observed in only one inactive case. After therapy, there was no 99mTc-tetrofosmin uptake in 3 patients, which correlated well with chest radiography and clinical findings. In the other 2 patients, 99mTc-tetrofosmin uptake was slightly decreased when compared with a previous scan that correlated with radiological and clinical findings. In 1 patient with bilateral lung disease, 99mTc-tetrofosmin uptake decreased on the right lung lesions, whereas the left lung lesions persisted with no change. The mean early and delayed L/NL ratios of APTB were 1.53+/-0.22 and 1.45+/-0.21, respectively. Although 99mTc-tetrofosmin uptake in APTB lesions was more visually marked in early images than that in delayed images, there was no statistically significant difference between these two sets of images. CONCLUSION: Technetium 99m-tetrofosmin scintigraphy showed increased uptake in APTB lesions related to disease activity. After treatment, 99mTc-tetrofosmin uptake disappeared or decreased, correlating well with radiological and clinical findings. Technetium 99m-tetrofosmin scintigraphy may have a complementary role in the assessment of APTB as well as in follow-up treatment. PMID- 9867154 TI - Coregistration of FDG PET and MRI of the head and neck using normal distribution of FDG. AB - For better localization of head and neck structures by PET with 2-(18)F-2-deoxy-D glucose (FDG), direct incorporation of anatomical information from MRI by the coregistration of FDG PET and MRI without external markers is proposed. METHODS: Seventeen patients with neoplasms and 16 normal subjects who had both FDG PET and MRI were studied. First, the three-dimensional normal distribution of FDG was evaluated, and then the structures of the head and neck regions with normal distribution patterns of FDG were used as internal markers for the coregistration of PET and MRI. The effectiveness of the coregistration was evaluated using focal neoplasms that were identified by both PET and MRI as fiducial internal markers. RESULTS: The normal structures selected as internal landmarks for coregistration were the tonsils, salivary glands, mucosal layers of the oral cavity and pharynx, spinal cord, inferior portion of the frontal lobe, cerebellum and nasal turbinates. These structures were more easily observed in sagittal or coronal sections than in transaxial sections. All primary neoplasms were delineated by PET, whereas 4 were missed by MRI. Thirteen primary tumors and 7 cervical lymph node metastases coregistered well, with a center-of-mass distance of <2 mm, whereas 10 lymph node metastases were slightly misregistered, with a center-of mass distance of 7.8+/-6.5 mm (mean+/-s.d.), probably due to differences in neck positions. CONCLUSION: Normal distribution of FDG uptake in the head and neck regions delineated by multidirectional sections is important for effective coregistration of FDG PET with MRI. PMID- 9867155 TI - Technetium-99m(V)-DMSA and gallium-67 in the assessment of bone and joint infection. AB - The aim of our study was to investigate the diagnostic value of scans with 99mTc(V)-dimercaptosuccinic acid (DMSA) to localize bone and joint infection compared with scans using 67Ga. METHODS: Thirty-six patients referred for investigation of bone and joint infection were studied. In all patients, a bone scan was obtained initially. Subsequently, comparative scans with 99mTc(V)-DMSA and 67Ga were performed 1 wk apart. Microbiological findings, pathologic findings and/or clinical follow-up (until symptoms disappeared) were considered to be proof of the presence of bone and joint infection. RESULTS: Technetium-99m (V) DMSA showed greater sensitivity and accuracy than 67Ga in the assessment of bone and joint infection, although the difference was not statistically significant. CONCLUSION: In comparison with a 67Ga scan, a 99mTc(V)-DMSA scan, in combination with a bone scan, is a reliable way to diagnose bone and joint infection. Both tracers were useful in the diagnosis of bone and joint infection. PMID- 9867157 TI - Gallium-67 scintigraphy to predict response to therapy in active lupus nephritis. AB - Gallium-67-citrate has been used to detect inflammation for decades, and 67Ga uptake usually indicates an active, potentially curable lesion. In this study, we determined the value of 67Ga renal scintigraphy for predicting response to therapy in patients with lupus nephritis. METHODS: Forty-seven patients with lupus nephritis and abnormal serum creatinine or elevated 24-hr urine protein were enrolled. Delayed 48-hr 67Ga imaging was performed to evaluate 67Ga uptake by the kidneys. Serum creatinine and 24-hr urine protein values were obtained at the beginning of this study and after 1 yr of treatment. Serum creatinine was considered abnormal at levels greater than or equal to 1.4 mg/dl and 24-hr urine protein at levels greater than or equal to 1.0 g/day. When the value of serum creatinine or 24-hr urine protein obtained 1 yr after treatment was in the normal range or was 50% of the initial abnormal value, the patient was considered to have good response to treatment. RESULTS: Gallium-67 renal scan showed good correlation with the response to therapy in patients with lupus nephritis. In the negative 67Ga scan group, no significant changes in laboratory data were noted between onset of this study and after 1 yr of therapy. In the positive 67Ga scan group, there were significant decreases in serum creatinine and 24-hr urine protein levels 1 yr after treatment, especially in 24-hr urine protein, with p values of 0.019 and 0.0007 respectively, by Student's t-test for dependent samples. Moreover, 11.5% of patients with a negative 67Ga scan had a good response to treatment, whereas 71.4% of patients with a positive 67Ga scan had a good response to treatment. CONCLUSION: We suggest that 67Ga renal scan is a valuable predictor of response to therapy in patients with lupus nephritis. PMID- 9867156 TI - Chromium-51-EDTA clearance in adults with a single-plasma sample. AB - In 1996, a committee on renal clearance recommended a mean sojourn time-based methodology for single-sample determination of plasma clearance of 99mTc diethylenetriamine pentaacetic acid (DTPA) to be used on adults if the patient's glomerular filtration rate (GFR) is suspected to be >30 ml/min. The main purpose of this study was to derive a mean sojourn time-based formula for calculation of 51Cr-ethylenediamine tetraacetic acid (EDTA) clearance in adults. METHODS: Two groups of patients with 51Cr-EDTA clearance (Cl) between 16 and 172 ml/min were studied. In Group I (n = 46), reference Cl was determined as a multiplasma sample, single-injection method (ClSM). Sixteen blood samples were drawn from 0 until 5 hr after a single intravenous injection of 51Cr-EDTA. In Group II (n = 1046), reference Cl was determined by the Brochner-Mortensen four-sample clearance method (ClBM). The plasma time-activity curves of Group I were used to derive two mean sojourn time-based formulas (Formulas 1 and 2) for calculation of a single-sample clearance. Formula 1 was derived from the entire time-activity curve, whereas the derivation of Formula 2 used only the final slope of the time activity curve. The accuracy of the two formulas and the Christensen and Groth 99mTc-DTPA formula was tested on Group II. RESULTS: Chromium-51-EDTA Cl calculated by Formula 1 was almost identical to the Cl calculated by the reference Cl method (r = 0.982; SDdiff = 5.82 ml/min). Both 51Cr-EDTA Cl calculated by Formula 2 and by the 99mTc-DTPA formula showed close correlation with the reference method (r = 0.976, r = 0.985, respectively) but systematically overestimated GFR for the whole range of clearance values by 3.5 and 3.2 ml/min (p<0.001), respectively. CONCLUSION: It is possible to get an accurate determination of 51Cr-EDTA Cl from a single-plasma sample in adults by the mean sojourn time methodology. The determination is marginally more accurate (p<0.001) if using a formula derived from the entire plasma time-activity curve than from only the final slope. The single-sample formula derived for determination of 99mTc-DTPA Cl tends slightly to overestimate GFR if used to calculate 51Cr-EDTA Cl. PMID- 9867158 TI - Scintigraphic localization of lymphatic leakage site after oral administration of iodine-123-IPPA. AB - Chylothorax can occur secondary to traumatic lesions of the thoracic duct caused by chest injuries, surgical procedures involving the pleural space, neoplasms or malformations of the lymphatics. METHODS: Lymphatic leakage sites were localized by scintigraphy after oral administration of the 123I-labeled long-chain fatty acid derivative iodophenyl pentadecanoic acid (IPPA). We report on three patients with different lymphatic leakage sites and on one normal control subject. RESULTS: IPPA scintigraphy localized the lymphatic leakage site correctly in all three patients. In two of them, the method even guided the successful surgical treatment of the leakage. CONCLUSION: This approach is suitable for detecting lymphatic leakages of intestinal origin. PMID- 9867159 TI - Fluorine-18-FDG PET and technetium-99m antigranulocyte antibody scintigraphy in chronic osteomyelitis. AB - The aim of this study was to assess the usefulness of PET with 2-18F-fluoro-2 deoxy-D-glucose (FDG), as compared to immunoscintigraphy (IS) with 99mTc-labeled monoclonal antigranulocyte antibodies (AGAbs), in the detection of chronic osteomyelitis. METHODS: Fifty-one patients suspected of having chronic osteomyelitis in the peripheral (n = 36) or central (n = 15) skeleton were evaluated prospectively with static FDG PET imaging and combined 99mTc-AGAb/99mTc methylene diphosphonate (MDP) bone scanning within 5 days. FDG PET and IS were evaluated in a blinded and independent manner by visual interpretation, which was graded on a five-point scale of two observers' confident diagnosis of osteomyelitis. Receiver operating characteristic (ROC) curve analysis was performed for both imaging modalities. The final diagnosis was established by means of bacteriologic culture of surgical specimens and histopathologic analysis (n = 31) or by biopsy and clinical follow-up over 2 yr (n = 20). RESULTS: Of 51 patients, 28 had osteomyelitis and 23 did not. According to the unanimous evaluation of both readers, FDG PET correctly identified 27 of the 28 positives and 22 of the 23 negatives (IS identified 15 of 28 positives and 17 of 23 negatives, respectively). The area under the ROC curve was 0.97/0.97 (reader 1/reader 2) for FDG PET and 0.87/0.90 for IS, with a high degree of interobserver concordance (K-values were 0.96 for FDG PET and 0.91 for IS). In the central skeleton, the ROC curve area was 0.98/1.00 for FDG PET and 0.71/0.77 for IS (p<0.05). On the basis of ROC analysis, the overall accuracies of FDG PET and IS in the detection of chronic osteomyelitis were 96%/96% and 82%/ 88%, respectively. With regard to the optimal threshold values, sensitivity and specificity were 100%/97% and 95%/95% with FDG PET, compared to 86%/92% and 77%/82% with IS, respectively. CONCLUSION: In the peripheral skeleton, both FDG PET and combined 99mTc-AGAb/99mTc-MDP scanning are appropriate imaging modalities to diagnose chronic osteomyelitis. FDG PET additionally allows reliable differentiation between osteomyelitis and infection of the surrounding soft tissue. In the central skeleton within active bone marrow, FDG PET is highly accurate and superior to AGAb imaging in the diagnosis of chronic osteomyelitis, which frequently presents as a nonspecific photopenic lesion at scintigraphy with labeled white blood cells. PMID- 9867160 TI - Renography before heart transplantation in patients with cardiomyopathy. AB - In patients with ischemic cardiomyopathy (CM), abnormal renograms may result not only from circulatory failure (which should reverse after transplantation) but also from intrinsic renal disease (which contraindicates heart transplantation). Here, the outcome of heart transplantation was related to preoperative renograms, and the differentiating and prognostic value of renography was analyzed. METHODS: The study population consisted of 50 patients with ischemic CM expecting heart transplantation. Anatomical renal pathology was excluded in all patients. Dynamic renal scintigraphy was performed with 99mTc-mercaptoacetyltriglycine. Background subtracted renograms were inspected visually and characterized numerically. Mean parenchymal transit time (mPTT), renal tracer content at 15 min (RTC15) and retention index (RI) were determined. The parametric renogram values were related to a normal reference group of 64 patients. The preoperative renograms were matched with the postoperative outcome. RESULTS: Three characteristic types of symmetrical findings in the kidneys were found: no pathological findings, mildly delayed peak and excretion phase and severely delayed peak and excretion phase. Pathological renograms were observed in 36 of 50 (72%) patients. The mean parametric renogram values in ischemic CM were as follows: Group A (normal kidney function), mPTT = 142+/-26.6 sec, RTC15 = 22.3%+/-4.6% and RI = 24.7+/-11.9; Group B (mild dysfunction), mPTT = 210+/-44.0 sec, RTC15 = 42.6%+/-10.3% and RI = 101.4+/-50.5; Group C (severe dysfunction), mPTT = 320+/-94.2 sec, RTC15 = 79.6%+/-15.9% and RI = 347.7+/-194.7; and reference patients (normal kidney function), mPTT = 137+/-31.1 sec, RTC15 = 22.8%+/-3.8% and RI = 24.6+/-7.9. Postoperative serum creatinine levels were <1.5 mg/dl in all Group A patients, between 1.5 and 2.5 mg/dl in 78% of Group B patients and >2.5 mg/dl in 75% of Group C patients. CONCLUSION: Renography revealed abnormal kidney function when structural pathology was excluded. The renographic abnormalities in ischemic CM did not reflect simply the circulatory failure. The numerical grading of renograms allowed patient stratification, suggestive of possible renal insufficiency after cardiac transplantation and immunosuppressive therapy. With further experience, renography may become a useful tool for predicting postoperative outcome in ischemic CM. PMID- 9867161 TI - Post-therapy iodine-131 localization in unsuspected large renal cyst: possible mechanisms. AB - Sensitive and specific, whole-body 131I scintigraphy remains an important technique for diagnosing metastases from differentiated papillary or follicular thyroid carcinoma. False-positive 131I localization is well recognized and can occur in a variety of conditions. We present a case of intense 131I localization in a previously unsuspected large renal cyst; the lesion was not visualized on routine preablation diagnostic 131I scintigraphy but was obvious on post therapeutic whole-body imaging, underscoring the value of post-therapy imaging in detecting abnormalities not apparent on diagnostic studies. Radioiodine within the urinary bladder or, at times, the renal collecting system is expected, because 131I excretion is primarily by glomerular filtration. In the case presented here, 131I activity within the renal cyst supports the concept that iodide is subject to an active secretory process by the renal tubule. PMID- 9867162 TI - Biodistribution and kinetics of holmium-166-chitosan complex (DW-166HC) in rats and mice. AB - The fate of 166Ho-chitosan complex, a radiopharmaceutical drug for cancer therapy, was determined by studying its absorption, distribution and excretion in rats and mice. METHODS: Holmium-166-chitosan complex [0.75 mg of Ho(NO3)3 x 5H2O and 1 mg chitosan/ head] was administered intrahepatically to male rats. Radioactive concentrations in blood, urinary and fecal excretion and radioactive distribution in tissues were examined. To determine the effects of chitosan in 166Ho-chitosan complex, 166Ho alone [0.75 mg of Ho(NO3)3 x 5H2O/head] was intrahepatically administered to male rats, and radioactive concentrations in blood, urinary and fecal excretion and radioactive distribution were examined. In B16 melanoma-transplanted nude mice, radioactive distribution after intratumoral administration of 166Ho-chitosan complex [0.075 mg of Ho(NO3)3 x 5H2O and 0.10 mg chitosan/head] was investigated also. RESULTS: After administration of 166Ho chitosan complex, the radioactive concentrations in blood were low, and cumulative urinary and fecal excretions over a period of 0-72 hr were 0.53% and 0.54%, respectively. The radioactive concentrations in tissues and the whole-body autoradiography images showed that most of the administered radioactivity was localized at the administration site, and only slight radioactivity was detected from the liver, spleen, lungs and bones. On the other hand, results of intrahepatic administration of 166Ho alone showed high radioactive concentrations in the blood, and the whole-body autoradiographs showed that the administered radioactivity was distributed in many organs and tissues. These results strongly suggest that 166Ho is retained at the administration site only when it forms a chelate complex with chitosan. Autoradiographs after intratumoral administration of 166Ho-chitosan complex showed that radioactivity was localized at the site of administration without distribution to the other organs and tissues. CONCLUSION: Administered 166Ho-chitosan complex is retained at the administration site after either intrahepatic or intratumoral administration to rats or tumor-transplanted nude mice. PMID- 9867163 TI - Absolute organ activity estimated by five different methods of background correction. AB - Accurate absorbed dose estimates in radionuclide therapy require patient-specific dosimetry. In patient-based dosimetry, estimation of absolute organ uptake is essential. The methods used should be reasonably accurate as well as easy to perform in routine clinical practice. One of the major sources of uncertainty in quantification of organ or tumor activity from planar images is the activity present in the tissue surrounding the source. METHODS: To estimate organ activity as a function of organ-to-background activity concentration ratio, a cylindrical phantom, filled with 5.6 liters of water was used to simulate the abdomen of a patient. Two other cylinders of 150 ml each, representing the kidneys, were each filled with 19 MBq 99mTC and were positioned in the abdomen phantom. The phantom was imaged with a dual-head gamma camera with the kidneys placed at posterior depths of 1-, 5- and 10-cm at kidney-to-background activity concentration ratios of infinity, 10:1, 5:1 and 2:1. The conjugate view geometric mean counting method was used to quantify activity. Five methods for background correction were applied: (1) no correction; (2) conventional background correction (simple subtraction of the background counting rate from the source region counting rate); (3) Kojima method (background corrected for organ thickness and depth); (4) Thomas method (analytical solution); and (5) Buijs method (background corrected for organ and total-body thickness). RESULTS: Since the results were identical for both kidneys, only the left kidney activity measurements are presented. The accuracy of the five background correction methods is given as the percentage difference between the actual and measured activity in the left kidney. For Method 1, the percentage difference ranged from 2% with an infinite kidney-to-background activity concentration ratio to +413% with a 2:1 ratio. For Method 2, these values ranged from -1% to -80%, for Method 3 from +11% to -18%, for Method 4 from -2% to +120% and for Method 5 from -4% to +39%. CONCLUSION: Even though quantitative SPECT is the most rigorous method for activity quantification in conditions of low organ-to-background activity concentration ratio, planar scintigraphy can be applied accurately if appropriate attention is paid to background correction. Using relatively simple background subtraction methods, the quantitative planar imaging technique can result in reasonably accurate activity estimates (Methods 3 and 5). The use of Kojima's method is preferable, especially at very low source-to-background activity concentration ratios. PMID- 9867164 TI - Technetium-99m-labeled liposomes to image experimental colitis in rabbits: comparison with technetium-99m-HMPAO-granulocytes and technetium-99m-HYNIC-IgG. AB - Scintigraphic techniques are routinely used for the evaluation of the extent and severity of inflammatory bowel disease. Currently, the radiopharmaceutical of choice is 99mTc-hexamethyl propyleneamine oxime (HMPAO)-leukocytes. We studied the imaging potential of two recently developed 99mTc-labeled agents, polyethylene glycol (PEG)-coated liposomes and hydrazinonicotinate (HYNIC) IgG, in a rabbit model of acute colitis, and compared them with that of 99mTc-labeled, granulocyte-enriched (>90%), white blood cells. METHODS: Acute colitis was induced in rabbits by retrograde instillation of trinitrobenzene sulfonic acid. After 48 hr, 37 MBq of each radiopharmaceutical was administered intravenously. Gamma camera images were taken at 0, 1, 2, 4, 10 and 24 hr. At 4 and 24 hr postinjection, groups of rabbits were killed, and the uptake of the radiolabel in the dissected tissues was determined. For each affected 5-cm segment, the colitis index (CI, affected-to-normal-colon-uptake ratio) was calculated and correlated to the macroscopically scored severity of inflammation. RESULTS: All three agents visualized the colitis lesions within 1 hr postinjection. The CI correlated with the severity of the abnormalities. With increasing severity, the CI at 4 hr postinjection for liposomes was 3.89+/-0.73, 4.41+/-0.47 and 5.76+/-0.65; for IgG 1.67+/-0.08, 3.92+/-0.44 and 6.14+/-0.65; and for granulocytes 2.90+/-0.09, 6.15+/-0.96 and 9.36+/-3.35. For liposomes, the CI further increased during 24-hr postinjection to 6.56+/-0.84, 8.50+/-0.53 and 10.61+/-1.34, respectively. The CI for the other two agents did not change significantly with time. CONCLUSION: In this rabbit model, 99mTc-labeled granulocytes, IgG and liposomes all rapidly visualized colonic inflammation. Granulocytes and liposomes showed the highest CI. Technetium-99m-labeled PEG-liposomes may be an attractive alternative for labeled leukocytes to image inflammatory bowel disease, because they can be prepared off the shelf and no handling of blood is required. PMID- 9867165 TI - Distribution of glutathione and technetium-99m-meso-HMPAO in normal and diethyl maleate-treated mouse brain mitochondria. AB - The aim of this study was to explain the contribution of mitochondria to the accumulation of 99mTc-meso-hexamethyl propyleneamine oxime (HMPAO) in the brain, after examinations were performed. METHODS: We studied subcellular distribution of 99mTc-meso-HMPAO and glutathione (GSH) in normal and diethyl maleate (DEM) administered mice. RESULTS: In normal brain, major radioactivity was found in the mitochondrial (49.0%) and cytosolic fractions (33.0%), while the GSH content was high in the cytosol (63.2%) and mitochondria (30.6%). The radioactivity in mitochondrial, cytosolic, microsomal and nuclear fractions was decreased in a dose-dependent manner by DEM, a GSH depleting agent, to 32.2% (mitochondrial) and 24.7% (cytosolic) of the control by a dose of 550 mg/kg. The GSH content in mitochondrial and cytosolic fractions also decreased in a dose-dependent manner on DEM treatment to 29.3% (mitochondrial) and 30.0% (cytosolic) of the control by 550 mg/kg of DEM. A good correlation was found between the uptake of 99mTc-meso HMPAO and GSH content in mitochondrial, cytosolic and nuclear fractions, with a correlation coefficient (r) of 0.814, 0.834 and 0.784, respectively. CONCLUSION: Mitochondria are a major subcellular fraction for the uptake of 99mTc-meso-HMPAO by the brain, and GSH in mitochondria contributes to the accumulation of 99mTc meso-HMPAO. PMID- 9867166 TI - Multiple line source array for SPECT transmission scans: simulation, phantom and patient studies. AB - Accurate attenuation and scatter corrections in quantitative SPECT studies require attenuation maps of the density distribution in the scanned object. These can be obtained from simultaneous emission/transmission scans. METHODS: A new method has been developed using a multiple line source array (MLA) for transmission scans, and its performance has been investigated using computer simulations and experimental data. The activity in the central lines of the MLA was higher than at the edges of the system, so that more transmission photons would be directed toward the thicker parts of the human body. A series of transmission-only and simultaneous emission/transmission studies were performed for different phantom configurations and human subjects. Attenuation maps were generated and used in reconstruction of attenuation-corrected emission images. RESULTS: The mu coefficients for attenuation maps obtained using the MLA system and simulated and experimental data display no artifacts and are qualitatively and quantitatively correct. For phantoms, the agreement between the measured and the true value of mu for water was found to be better than 4%. The attenuation corrected emission images for the phantom studies demonstrate that the activity in the heart can be accurately reconstructed. A significant qualitative improvement was also obtained when the attenuation correction was used on patient data. CONCLUSION: Our results indicate that the MLA transmission source can be used in simultaneous transmission/emission imaging to generate accurate attenuation maps. These maps allow for performing an object-specific, attenuation correction of the emission images. PMID- 9867167 TI - Ghost-like scintigraphy. PMID- 9867168 TI - Prevalence of adverse reactions to positron emitting radiopharmaceuticals in nuclear medicine. Pharmacopeia Committee of the Society of Nuclear Medicine. AB - This study was undertaken to determine the prevalence of adverse reactions to positron emitting radiopharmaceuticals as well as to nonradioactive drugs used in interventional nuclear medicine during PET studies. METHODS: A prospective 4-yr study was performed with 22 collaborating institutions using a questionnaire, which indicated for each month of the study the number of PET procedures performed, the number of adverse reactions to PET radiopharmaceuticals as well as the number of adverse reactions to interventional nonradioactive pharmaceuticals used for PET. RESULTS: A total of 33,925 radiopharmaceutical doses were recorded in a retrospective examination of records by the 22 participating institutions. In addition, the total prospective number of administered doses recorded by the participants was 47,876, for a total number of positron emitting radiopharmaceutical administrations of 81,801. No adverse reactions were found from any PET radiopharmaceutical dose. There were no deaths or hospitalizations caused by nonradioactive interventional pharmaceuticals used adjunctive to PET studies. CONCLUSION: PET radiopharmaceuticals have an extraordinary safety record with no adverse reactions reported in over 80,000 administered doses in this study. PMID- 9867169 TI - Things are perhaps not quite so simple... PMID- 9867170 TI - What is a false-positive somatostatin receptor scintigraphy? PMID- 9867171 TI - Costs versus charges. PMID- 9867172 TI - Concerns about risks of irradiation during pregnancy. PMID- 9867173 TI - Failure to detect drainage to the popliteal and epitrochlear lymph nodes on cutaneous lymphoscintigraphy in melanoma patients. PMID- 9867174 TI - Somatostatin receptor scintigraphy in small-cell lung cancer. PMID- 9867175 TI - Role of health economics in nuclear medicine. PMID- 9867176 TI - Sigmoidal curves. PMID- 9867177 TI - Scoliosis circa 2000: radiologic imaging perspective. I. Diagnosis and pretreatment evaluation. AB - Plain film imaging remains important for the diagnosis and surveillance of scoliosis, as well as for the detection of complications after surgery. Advances in CT and MR imaging have greatly improved the ability to detect or confirming non-idiopathic causes of scoliosis, including abnormalities within the spinal canal. Three-dimensional thinking has become more important in evaluating and understanding scoliosis. PMID- 9867178 TI - Use of contrast-enhanced MR imaging to detect sacroiliitis in children. AB - PURPOSE: To verify the diagnostic value of contrast-enhanced MR imaging compared with conventional radiography in the diagnosis of sacroiliitis in children. DESIGN AND PATIENTS: Radiography and MR imaging of the sacroiliac joints were performed in 185 children subdivided into the following groups according to the modified European Spondyloarthropathy (SpA) Study Group (ESSG) criteria: group 1, undifferentiated spondyloarthropathy (uSpA) (n=53, 94.5% HLA-B27+); group 2, differentiated SpA (n=45, 93.3% HLA-B27+); group 3, patients with no signs of SpA other than oligoarthritis (n=39, 92.3% HLA-B27+); group 4, HLA-B27+ controls with various other non-SpA diagnoses (n=22); and group 5, HLA-B27-controls with various other non-SpA diagnoses (n=26). Radiographs were evaluated on the basis of the modified New York criteria independently by three experienced radiologists masked to the clinical data. In a second step, the same radiologists independently evaluated the MR images without knowledge of the clinical data and radiographic findings using the recently published criteria developed by our group. These criteria allow differentiation of acute and chronic inflammatory changes. RESULTS: Radiography demonstrated sacroiliitis in 18 patients: 4 of 53 in group 1 (7.5%), 14 of 45 in group 2 (31%), but none in groups 3, 4 and 5. In contrast, MR imaging demonstrated acute and/or chronic sacroiliitis in 44 patients: 18 of 53 in group 1 (34%), 21 of 45 in group 2 (46.7%) and 5 of 39 in group 3 (12.8%), but none in groups 4 and 5. The percentage of sacroiliitis detected by MR imaging was significantly higher than that detected by radiography (P<0.001). CONCLUSION: Contrast-enhanced MR imaging is a useful method for detecting sacroiliitis in children. Advantages of contrast-enhanced MR imaging compared with conventional radiography are a higher sensitivity due to the ability to document early and acute changes and the absence of radiation exposure. PMID- 9867180 TI - Chondrosarcoma of the short tubular bones of the hands and feet. AB - OBJECTIVE: To report 12 cases of chondrosarcoma in a rare location, the short tubular bones of the hands and feet, as well as 12 cases of enchondroma in similar locations, emphasizing the radiologic and histopathologic features. DESIGN AND PATIENTS: All relevant cases that had both histologic slides and radiographs available were taken from the files of one orthopedic referral hospital and the personal files of one of the authors. A similar number of enchondroma cases was selected at random from the files. RESULTS AND CONCLUSIONS: One malignancy arose in a background of enchondromatosis, with all the others being solitary lesions. A pathologic diagnosis of malignancy is often difficult in the absence of radiologic signs of malignancy (cortical destruction with or without soft tissue extension). However, three cases were unusual in that the initial radiograph demonstrated a benign appearance. Another group of three malignancies was surprisingly indolent biologically. The treatment of choice is ray resection (or more limited amputation in a lesion of the middle or distal phalanx). PMID- 9867179 TI - Closed ruptures of the flexor digitorum tendons: MRI evaluation. AB - OBJECTIVE: To assess the MRI findings in cases of closed rupture of the flexor digitorum tendons (FDT). PATIENTS AND DESIGN: Ten patients with a clinical suspicion of rupture of FDT underwent MRI before surgery. None of the patients presented a skin injury. Fingers were imaged using axial T1-weighted SE sequences, three-dimensional GE images, and curved reconstructions. RESULTS: Twelve FDT had surgical confirmation of rupture. Flexor digitorum profundus (FDP) and flexor pollicis longus (FPL) tendons were more frequently ruptured (n=8) than flexor digitorum superficialis (FDS) tendons (n=4). MR images accurately depicted the level of the rupture. The gap between the tendon ends (mean 45 mm, range 21 70 mm) was assessed best with curved reconstructions and was well correlated with the surgical findings. The proximal end mainly retracted into the palm or the carpal tunnel (n=8), and less frequently into the digital canal (n=4). In two cases, the proximal end curled up in the palm, clinically simulating a rupture of a lumbrical muscle in one case. MRI also showed the appearance of the adjacent tendons. CONCLUSION: MRI accurately depicted the level of rupture and the gap between the tendon ends, which assisted the surgical choice between suture, graft or tendon transfer. PMID- 9867181 TI - Sclerotome distribution of melorheostosis and multicentric fibromatosis. AB - A case of melorheostosis affecting the C7 sclerotome is described, in association with synchronous multicentric fibromatosis. The foci of fibromatosis also affected the C7 sclerotome, suggesting a similar underlying pathogenesis for both diseases. PMID- 9867182 TI - Castleman's disease of the erector spinae muscle. AB - Castleman's disease, angio-follicular lymph node hyperplasia, is a relatively rare benign tumour. It typically arises in the mediastinum. We report a unique case arising in the erector spinae muscle. The case demonstrates the importance of radiological investigation and embolisation prior to obtaining a tissue diagnosis and subsequent surgical excision. PMID- 9867183 TI - Primary multifocal tuberculous osteomyelitis with involvement of the ribs. AB - Two cases of primary multifocal tuberculous osteomyelitis with involvement of the rib cage are presented. The lungs were normal and the appearance of the skeletal lesions did not suggest tuberculosis. These lesions were predominantly lytic, with minimal soft tissue involvement. Tuberculosis should be high in the differential diagnosis of multiple destructive bone lesions, especially in patients from regions where tuberculosis is endemic. PMID- 9867184 TI - Florid periosteal reaction and focal fibrocartilaginous dysplasia. AB - Focal fibrocartilaginous dysplasia (FFCD) is a rare condition causing tibia vara in childhood. It is characterized by progressive tibia vara in young children with a characteristic radiographic lesion. This paper is thought to be the first to describe FFCD exhibiting florid periosteal reaction at the time of presentation with a subtle faint osteolytic lesion in the diametaphysis of the proximal tibia. PMID- 9867185 TI - Parosteal osteoma of the iliac bone. PMID- 9867186 TI - Morphological, immunohistochemical and autoradiographic studies of thyroid autonomy. AB - The values and limits of morphological, immunohistochemical and autoradiographic methods in studies of thyroid autonomy are briefly discussed. For meaningful studies of molecular aspects of thyroid autonomy--such as for example TSH receptor and Gs-alpha gene mutations--it is absolutely crucial that the tissue analysed is well characterized and really is autonomous. This is particularly important in view of the well known heterogeneity of human goiter tissue in respect to many if not all functional and proliferative parameters. To prove functional and proliferative autonomy of thyroid tissue, autoradiography is a very helpful tool, while simple morphology and immunohistochemistry do not contribute substantially to this aim. PMID- 9867187 TI - Activation of the cAMP cascade in thyroid autonomy. PMID- 9867188 TI - Pathogenesis of toxic thyroid adenomas and nodules: relevance of activating mutations in the TSH-receptor and Gs-alpha gene, the possible role of iodine deficiency and secondary and TSH-independent molecular mechanisms. AB - In iodine deficiency areas, activating mutations in the TSH receptor and Gs-alpha gene are found in the majority of toxic thyroid adenomas and in some toxic goiter nodules. Since TSH receptor and Gs-alpha gene mutations are very rare in areas with high iodine supply, iodine deficiency has been suspected to favor the occurrence of these mutations by yet unknown pathways. However, TSH receptor and Gs-alpha gene mutations alone are not sufficient to cause toxic adenomas and nodules. There is compelling evidence that other secondary and cAMP-independent mechanisms, including enhanced expression of various growth factors, their corresponding receptors and of signaling proteins, may affect the mutated cell and thus promote cell proliferation and in turn generation of the tumor. PMID- 9867189 TI - Autosomal dominant nonautoimmune hyperthyroidism. Clinical features-diagnosis therapy. AB - Autosomal dominant nonautoimmune hyperthyroidism is a hereditary form of hyperthyroidism caused by constitutively activating germline mutations in the TSH receptor gene. Clinical features comprise familial prevalence of thyroid autonomy in more than 2 generations and conditions of persisting neonatal hyperthyroidism or nonautoimmune hyperthyroidism of childhood onset with frequent relapses of hyperthyroidism under thyrostatic therapy and after thyroid surgery. Once clinically suspected the diagnosis can be confirmed by mutation analysis of genomic DNA extracted from a routinely obtainable EDTA blood sample. In patients with hereditary nonautoimmune hyperthyroidism a near total thyroidectomy is recommended as the first line treatment to avoid relapses from residual thyroid tissue with the activating TSHR mutation. Furthermore, genetic counselling of the affected patients is advised. PMID- 9867190 TI - Epidemiology of functional autonomy. AB - About 90% of all functional thyroid autonomies (FTA) are euthyroid for a prolonged period of time. It is estimated that more than 10% of goiter patients in iodine deficient regions and less than 10% in iodine rich areas have evidence of FTA. After the age of 40, the risk of hyperthyroidism decompensation gradually increases. This risk rises with increasing thyroid volume, nodularity and patient age. In the elderly, hyperthyroidism also occurs in the absence of goiter. After decades of iodine deficiency, especially the intake of unphysiologically high iodine concentrations may result in increased frequencies of hyperthyroidism. In iodine deficient regions, almost half of all cases of hyperthyroidism are FTA related. Following elimination of iodine deficiency, the rate of hyperthyroidism may be reduced below 10%. This will not affect the prevalence of immunogenic hyperthyroidism. The most reliable evidence of FTA is produced using the TcTU supp. test. The highly sensitive TSH0 and the TRH test are 2.5 times less sensitive. Thus, they may still indicate euthyroidism in scintigraphically compensated or decompensated FTA. The TSH0 screening is only recommended with a view to an improved cost/benefit ratio in the elderly, females above the age of 40, and ill persons. Acutely ill and psychiatric patients should be excluded. Already 1 year after the introduction of iodine into the medical treatment of thyroid disorders, Coindet reported in 1821 his epidemiologically relevant clinical observation of an increase in hyperthyroidism, predominantly of the functional autonomy type. In the meantime, detailed and universally accepted knowledge has become available on the pathogenesis and pathophysiology of functional thyroid autonomy (Gerber et al., 1985). Data on the epidemiology of functional autonomy continue to apply only to the regional population they are based on. They allow to draw conclusions on the prevalence and natural course of functional thyroid autonomy (FTA). The different forms and prevalence rates of hyperthyroidism reflect the severity and duration of the nutritional iodine deficiency on one hand and the quality of iodine prophylaxis on the other. PMID- 9867191 TI - Evaluation of risk in autonomously functioning thyroid nodules. AB - Based on iodine deficiency in Germany autonomous functioning nodules develop and are loaded with the risk of hyperthyroidism. Even subclinical hyperthyroidism has health implications including general symptoms, diseases of the cardiovascular system (atrial fibrillation, risk of thrombembolism, stroke and other embolic events) and a reduction of axial bone mineral density. Fortunately functional nodules have a very low probability of being malignant. However, there exist a coexistence of thyroid carcinoma outside the hot nodule. PMID- 9867192 TI - Course of thyroid nodules. AB - Thyroid nodules are present in up to 30% of the German population. The causative role of iodine deficiency which is still endemic in this country has long been established. Recent progress has shed some light on the pathogenesis of nodular thyroid disease which still remains less well understood than goitrogenesis. Most thyroid nodules appear to be of clonal origin. Functional abnormalities have been related to alterations within the TSH signaling cascade, particularly mutations in the TSH receptor and stimulating G-protein-alpha-subunit. Proliferation which is dissociable from thyroid function has been linked to genetic differences of the thyroid cells themselves and growth factors being partly overexpressed by thyroid nodules. Data regarding the correlation of the molecular characteristics to the clinical behavior and growth potential have not yet been elucidated. On the other hand, there are only a few clinical studies that have addressed the long-term natural history of thyroid nodules. From these studies at least it appears that thyroid nodules tend to grow slowly and their increase in size may even by modern ultrasonography technique become apparent only after several years. Those in vitro and in vivo observations have important implications for the planning of therapeutical trials. Studies have focused so far mainly on short term effects of different therapeutic regimens such as iodine or levothyroxine. However, pathophysiological considerations and clinical observation would encourage studies over more prolonged periods of time. PMID- 9867193 TI - Utility of third generation thyrotropin assays in thyroid function testing. AB - Determination of thyrotropin (TSH) by sensitive immunometric assays is currently judged as the most sensitive and also most cost-effective first-line approach to thyroid function testing. Further improvement of assay sensitivity has led to the description of third generation TSH assays with a functional sensitivity in the range of 0.01 to 0.02 mU/l. In the present study, we analyzed interassay precision profiles of a commercially available third generation assay (ACS:180 TSH-3) and documented the critical role of the time span used for the assessment of a method's functional sensitivity. By using a standardized approach with five serum pools measured in 30 different runs across a 6-week period, functional sensitivity was calculated as 0.015 mU/l. The TSH concentrations measured by two different third generation assays (ACS: 180 TSH-3 and Elecsys TSH) in samples from healthy blood donors were highly correlated (r = 0.76, n = 252). In some samples, however, discordant results were obtained. Euthyroid reference intervals were determined as 0.30-3.68 mU/l for the ACS:180 TSH-3 assay and as 0.36-3.64 mU/l for the Elecsys TSH assay. Reevaluation of reference intervals including only TPOAb or TgAb negative samples resulted in almost the same reference ranges. Measuring TSH concentrations in various patient populations, third generation assay turned out to be advantageous in the following clinical situations. (a) In patients with mildly suppressed but well detectable TSH concentrations due to functional thyroid autonomy (0.03-0.3 mU/l), overt hyperthyroidism can be excluded by third generation TSH measurement alone without the need of additional thyroid hormone measurements; (b) in patients receiving long term suppressive T4 treatment after thyroidectomy for differentiated thyroid cancer, measurement of basal TSH by third generation assays allow accurate monitoring of hormone therapy without the need for TRH testing; (c) in most patients with severe nonthyroidal illnesses and decreased TSH levels, TSH concentrations measured by third generation assays are only moderately suppressed and can be clearly discriminated from undetectable levels in overt hyperthyroidism. In conclusion, the use of third generation TSH assays is recommended in specialized clinical laboratories frequently analyzing samples taken in one of those clinical situations. PMID- 9867194 TI - Methods for measuring iodine in urine and serum. AB - The most important information in the determination of the status of iodine nutrition of a population comes from the measurement of the urinary excretion of iodine. Several methods are available for measuring urinary iodine. The choice among methods depends on the intended application, the number of samples, the cost and the technical capability. Epidemiological field studies demand simple, rapid and cost-effective methods. Suitable for these applications are the rapid urinary iodide test and the ammonium persulfate oxidation method which gives comparable results to the chloric acid method without having the drawbacks of being hazardous and explosive. In research studies however, sophisticated automated technology like the Technicon Autoanalyzer or Paired-Ion Reversed Phase HPLC are used in which the high cost of instrumentation are outweighed by the benefits of processing a large number of samples with high accuracy and minimal technician time. For determining serum inorganic iodide (SII) the HPLC assay is the method of choice, because contaminations from the protein bound iodine fraction do not interfere with the detection process. The clinical relevance of the measurement of SII is limited, but allows the calculation of the absolute iodine uptake which has great value in pathophysiologic studies. PMID- 9867195 TI - Role of ultrasound in the diagnosis of thyroid autonomy. AB - The role and the different technical modes of ultrasound in thyroid autonomy are discussed. B-mode sonography is routinely employed to e.g. calculate thyroid and nodule volume, describe the echo pattern of both the thyroid gland and focal lesions and to visualize neighboring organs such as lymph nodes. Sonography thus possesses a complementary role to scintigraphy, since it may further aid in the localization of toxic adenomas, in particular when they are dorsally located or it may change the therapeutic decision. An example for the latter would be the preference of a surgical approach in case of cystic degeneration of an autonomously functioning nodule. Recently, color-coded doppler sonography using color flow mapping has been employed. With the advent of this technique it has become possible to describe the pattern of vascularization in thyroid focal lesions such as rim perfusion or internal hypervascularization. This technique harbors a high sensitivity for toxic adenoma exhibiting high internal blood flow, but does not appear to reliably discriminate between benign and malignant lesions. Nevertheless, further improvement of this technique may be promising in further characterizing the functional status of sonographically detected lesions and in the prognostic assessment of post-treatment outcome. PMID- 9867196 TI - Scintigraphic evaluation of functional thyroidal autonomy. AB - The scintigraphic evaluation of thyroidal autonomy is performed both as a quantitative and qualitative thyroid scintigraphy with 99m-Tc-pertechnetate (99m TcO4-), using a gamma camera fitted with an on-line computer system. A strong and linear correlation between the global 99m-Tc-pertechnetate thyroid uptake (TCTU) and I-123 clearance has been recognised. Therefore TCTU-values can be used as an reliable equivalent of the iodine clearance in the evaluation of actual thyroid function. The clinical value of the TCTU in the diagnosis of thyroidal autonomy is limited because it represents iodine clearance of both normal and autonomous tissue. As a consequence scintigraphic diagnosis and quantification of autonomy can only be established if the global 99m-Tc-pertechnetate thyroid uptake under suppression (TCTUs) is determined. This method is valuable in risk stratification of spontaneous or iodine induced hyperthyroidism, in the estimation of the target volume prior to radioiodine therapy and in the evaluation of therapeutic success after definitive therapy. PMID- 9867197 TI - Medical therapy of thyroid autonomy. PMID- 9867198 TI - Treatment of hyperfunctioning thyroid nodules with percutaneous ethanol injection: Eight years' experience. AB - The aim of our study was to define the long-term efficacy and safety of percutaneous ethanol injection (PEI) for the treatment of autonomous thyroid nodule (ATN), and to optimise the clinical usefulness of such a therapy. We treated 132 patients with ATN (30 M and 102 F, aged 47.5+/-12.9 years; mean+/ SD), in case other established treatments were refused or contraindicated. Eighty five patients were affected by toxic adenoma and 47 suffered from pre-toxic nodules. Ethanol was administered weekly under sonographic control, in 7 sessions (range 2-16). During PEI treatment, 26 toxic elderly patients were treated with methimazole and propranolol. Three possible outcomes were identified for statistical analysis: failure (persistent suppression of extra nodular tissue uptake, along with elevated free thyroid hormone and undetectable TSH levels); partial cure (normal free thyroid hormone and low/undetectable TSH levels); complete cure (normal thyroid hormone and TSH levels; restored extra nodular uptake). The patients were followed for up to 8.5 years (median 76 months). PEI therapy was well tolerated by all patients though a mild to moderate local pain occurred in about 30% of sessions. Complete cure was achieved in all pre-toxic patients and in 60 (70.6%) patients with toxic adenoma, while partial cure was observed in 11 cases (12.9%) and failure in 14 (16.5%). A significant shrinkage of nodule volume was observed in all patients (p = 0.0001), while those with toxic nodules larger than 30 mL showed a significantly lower response rate to PEI (p < 0.05). At controls, only one patient developed subclinical hypothyroidism while, among partially cured patients, five relapsed. The administration of methimazole and/or propranolol did not modify PEI outcome. In conclusion, we suggest that PEI therapy may be the treatment of choice in patients with pre toxic thyroid adenoma where therapy is least necessary- despite the nodule volume. Though ethanol injection therapy of toxic thyroid nodules may be troublesome for the need of multiple sessions, it appears an effective alternative procedure in patients at poor surgical risk, and in younger patients in whom radioiodine is contraindicated. Since a special technical skill in intervention procedures is required, PEI therapy may be suitable only for patients living nearby a trained centre. PMID- 9867199 TI - Ultrasound-guided percutaneous ethanol injection in the treatment of autonomous thyroid nodules--a review. AB - Percutaneous ethanol injection therapy (PEIT) in patients with autonomously functioning thyroid nodules (AFTN) has been evaluated with respect to indication for PEIT, treatment procedure, results of therapy and side effects of this treatment. As PEIT has not been tested against the standard procedure of radioiodine treatment and surgery up to now, PEIT should be an alternative treatment for selected patients who cannot or will not be treated by radioiodine therapy or surgery. The clinical data available demonstrate that better results are obtained in patients with euthyroid AFTN or AFTN with subclinical hyperthyroidism as compared to patients with AFTN and overt hyperthyroidism, particularly in small thyroid nodules (thyroid nodular volume less than 15 ml). After PEIT significant reduction of thyroid nodular volume can be achieved (reduction 21 to 88 %). Adverse events of PEIT are few and usually mild and with special respect to transient dysphonia, strongly correlated with the technical skills of the operator. Limitations of PEIT are mainly the need of repeat ethanol injections in order to achieve a complete cure of AFTN. In conclusion, a prospective controlled randomized clinical study has to be carried out in patients with AFTN in order to define the role of PEIT in comparison to the standard therapies (radioiodine treatment, surgery), in the treatment of patients with AFTN conclusively. PMID- 9867200 TI - Radioiodine treatment of Plummer's disease. AB - The presented overview summarizes the results of radioiodine treatment in patients with different forms of Plummer's disease. From the beginning the outcome defined as elimination of autonomy was excellent in the unifocal autonomous adenomas, but unsatisfactory in multifocal and disseminated autonomy using a fixed dose concept. The "dosimetric compromise" (= reducing the dose and defining the total gland as the target volume) yielded in these two variants of autonomy satisfactory results comparable to those of the unifocal form, since the dose was chosen according to the amount of functional autonomy measured by the Tc 99m-uptake in the thyroid gland under TSH-suppression. PMID- 9867201 TI - Treatment of toxic nodular goitres: Comparative costing of radioiodine therapy and surgery. AB - Cost accounting has shown that the volume of tissue to be treated is the decisive factor in determining the cost of radioiodine therapy (RAITh). In the case of large goitres, the costs of excision (5.185 DM) and radioiodine therapy (5.562 DM) are, to a large extent, equivalent. Under the 1993 regulations for radiation protection, RAITh was cost-effective for treatment of toxic multinodular goitres up to volumes of 57 ml. However, new maximum permissible levels of radioactivity on discharge from hospital (250 MBq iodine-131 residual activity) have raised this threshold volume to 90 ml. In Germany, remuneration for a goitre resection is calculated from standard charges for total treatment without any consideration of the size and spectrum of medical services offered by different clinics, while remuneration for RAITh comes from payments for basic and specific, departmental medical services. University clinics with departments of nuclear medicine have relatively high basic costs. In the first quarter of 1998, the length of hospitalisation after RAITh (for all indications combined) was 4.6 days in university hospitals in Germany. Owing to this shorter length of hospitalisation, the payments received in some clinics fell far short of the total costs of this treatment calculated by cost accounting. PMID- 9867202 TI - Cost analysis of surgical procedures for toxic nodular goiter. AB - Effective by January 1, 1993, the Federal Care Rate Regulation (Bundespflegesatzverordnung; BPflV) has started increased competition between hospitals by introducing performance related reimbursements. All three categories of flat payments, Fallpauschale (FP), Sonderentgelt (SE) and Budget-patient (BP), apply to toxic nodular goiter surgery, governed solely by the extent of thyroid resection. Unlike FP 2.01 and FP 2.02 which do not cover all surgical expenses, the BP flat rate provides a comparatively generous refund for total thyroidectomy. With the oncoming reduction of upper values for residual radioactivity, hospitalization will fall after radioiodine therapy, resulting in lower overall refunds. With this anticipated fall in reimbursement, refunds for radioactive therapy must be increased in the near future to ensure adequate financial compensation. Economic arguments for either therapeutic option will then cease to play a major role, allowing medical therapeutic considerations to prevail. PMID- 9867203 TI - Surgical aspects of thyroid nodules previously treated by ethanol injection. AB - Some concern has been expressed about surgical operations in thyroid nodules previously treated by ethanol injection: the reasons are mainly represented both by the possibility of more surgical risks due to the adhesions caused by ethanol and the difficulty of interpreting the histological pattern when a cancer is suspected. During the last 8 years 219 patients underwent ethanol injection: among these 6 were subsequently submitted to surgical operation. Total thyroidectomy was performed in 5 cases and isthmusectomy in one case: No vocal cord palsy was registered in these patients.. They were all normocalcemic after surgery. Histology showed no significant fibrosis in the ethanol treated nodules but only macrofollicles and this did not affect the histological examination in presence of suspected malignancies. PMID- 9867204 TI - Differentiated operative therapy of thyroid autonomy (Plummer's disease). AB - The operative therapy of the autonomous thyroid deals almost exclusively with nodular goiters. There are only rare situations with purely diffuse autonomy in surgical patients. The endpoint of operative therapy is permanent elimination of clinically relevant autonomous function and thereby irreversible abolition of thyrotoxicosis, even in their latent form. Clinically relevant autonomous function normally ist linked to nodular structures of different size and different distribution, so that this aim corresponds automatically with the aim of complete removal of nodular structures, both in autonomous and in non autonomous goiters. Function is best preserved by leaving a homogenous remnant of considerable size. In different particular clinical situations (for example pregnancy, suspicion of malignancy, recurrent goiter, intrathoracic goiter, thyrotoxic crisis, Marine-Lenhart-syndrome) the basic principal of operative strategy is varied according to the respective situations. Unsatisfactory operative results are mainly caused by incomplete removal of nodules, based on insufficient surgical performance of morphological and functional diagnostics, which is related to operative uniformity. An operative strategy, which ist "fitted to morphology" and "regarding function" and which we call "selective strategy", in our opinion ist highly appropriate, to avoid remnant nodules and remnant autonomy and to preserve an normal remnant, even in different position and of different size. This selective surgery ist applicable both to autonomous and non-autonomous goiter. Compared with the classic subtotal, uniform procedure the incidence of remnant nodules is reduced from about 50% to about 5 %. Remnant autonomy is almost excluded, when mistakes are avoided (about 1%). The incidence of recurrent goiter and recurrent thyrotoxicosis is lowered to under 5%--but up to now there are too few reliable long-term follow-up studies. The selective surgery strategy demands flexibility regarding operative tactics, which can be simply classified into 5 basic situations, which we relate to the operative procedure per thyroid lobe. It requires experience and competence in carefully dissecting and--when necessary--manipulating the recurrent laryngeal nerve und the parathyroid glands. Under these conditions it is followed by a comparatively low rate of complications (permanent recurrent laryngeal nerve paralysis and hypoparathyroidism under 1%, respectively). Thus, the old dilemma of thyroid surgery can be solved, which consists of radical operation with higher morbidity and lower frequency of recurrent disease on the one hand and more limited operative procedures with fewer complications but more frequent recurrencies on the other hand. PMID- 9867205 TI - Substitution therapy after surgery for autonomous adenomas. AB - A substitution therapy with L-thyroxine subsequent to surgery on the thyroid gland due to autonomous dysfunction depends in particular on the extent of resection. A specific postoperative therapy is not necessary in areas with sufficient iodine supply if the remaining part of the thyroid gland guarantees a euthyroid metabolic state. The radicalness of the intervention is dependent on the existence of an unifocal, multifocal or disseminated autonomy. Surgery due to disseminated autonomy always necessitates a substitutive therapy with thyroid hormones and in most cases surgery due to multifocal autonomies require the same treatment. The therapy is initiated with a dose of 1.5 microg L-thyroxine per kg body weight. Suppression of the TSH level is not necessary. Due to the greater risk of recurrence the L-thyroxine administration should be complemented in areas of iodine deficiency with approximately 100-200 mg iodide. After operations with functionally adequate thyroid remnants (8-10 ml), an exclusive prophylaxis with 200 mg iodide can be implemented. The result of surgery should be sonographically documented three months after the intervention. PMID- 9867206 TI - Prospects for gene therapy of insulin-dependent diabetes mellitus. AB - The application of gene therapy to Type I (insulin-dependent) diabetes mellitus awaits improvements in gene transfer technologies and the development of better tools for accurate diagnosis of pre-diabetic people. Identification of the most promising candidate genes for gene transfer requires further elucidation of the molecular events involved in beta-cell autoimmune destruction, islet ontogeny and differentiation, and beta-cell function. This review outlines a number of possible targets for gene therapy in Type I diabetes, which could help prevent the autoimmune damage to islets, induce islet regeneration, and restore insulin production through engineering of self non-beta cells or beta-cell transplantation. It also evaluates their potential merits and drawbacks. PMID- 9867207 TI - Possible modulation by endothelin-1, nitric oxide, prostaglandin I2 and thromboxane A2 of vasoconstriction induced by an alpha-agonist in mesenteric arterial bed from diabetic rats. AB - We hypothesized that in diabetes arterial reactivity to constrictors is attenuated by certain endothelium-derived substances. We examined the vasoconstriction induced by methoxamine (alpha1-agonist) in isolated mesenteric arterial beds from streptozotocin (STZ)-induced diabetic rats and age-matched control rats. The dose-response curve for methoxamine was shifted to the right and the maximum contractile response was impaired in mesenteric arterial beds from diabetic rats. The methoxamine vasoconstriction was reduced in endothelium denuded preparations from controls rats, but increased in those from diabetic rats. Treatment with the nitric oxide synthase inhibitor NG-nitro-L-arginine enhanced the vasoconstrictions induced by methoxamine in both control and diabetic rats. Indomethacin had no effect on the methoxamine vasoconstriction in control rats, but it shifted the dose-response curve to the left in diabetic rats. Whether given with or without indomethacin, BQ-123, (an ET(A)-receptor antagonist) plus BQ-788 (an ET(B)-receptor antagonist) shifted the dose-response curve for methoxamine to the right in control rats (while reducing the maximum response) but to the left in diabetic rats. The methoxamine-stimulated release of 6-keto-prostaglandin F1alpha from the mesenteric arterial bed in diabetic rats was approximately four times that seen in the control rats, while the methoxamine induced release of thromboxane B2 (TXB2), a metabolite of thromboxane A2 (TXA2), was less in diabetic rats than in the control animals. These results suggest that an increased production of prostaglandin I2 (PGI2) and decreased formation of TXA2 could be responsible for the attenuation of the methoxamine-induced mesenteric vasoconstriction seen in diabetic rats, and these changes in the diabetic state could be partly responsible for the lower blood pressure seen in our diabetic rats. PMID- 9867208 TI - Depletion of nitric oxide synthase-containing neurons in the diabetic retina: reversal by aminoguanidine. AB - A close association of neuronal nitric oxide synthase-immunoreactive (nNOS-IR) neurons with the retinal vasculature has been reported and it is proposed that activation of these neurons could be the mechanism by which retinal blood flow and metabolism are linked. Further, advanced glycation end products (AGEs) have previously been shown to be increased in the diabetic retina and aminoguanidine (AG), an inhibitor of advanced glycation, has been shown to attenuate the development of AGE accumulation as well as the progression of experimental diabetic retinopathy. This study examined the effects of short (1 and 3 weeks) and long term (32 weeks) diabetes on nNOS-containing neurons of the retina using NADPH diaphorase (NADPHd) histochemistry. In addition, the effect of aminoguanidine (an inhibitor of advanced glycation and NOS) and NG-nitro-L arginine methyl ester (L-NAME) (a non-selective NOS inhibitor) on retinal nNOS containing neurons was examined in short and long term control and diabetic rats. In a separate study, the effect of 2,3 diamino-phenazine (NN0028) (an inhibitor of advanced glycation, but not NOS) was examined in short term (3 weeks) diabetic rats. The number of NADPHd-positive neurons per retina was reduced after one week of diabetes and remained decreased in long term diabetic rats, an effect not observed in diabetic rats rendered euglycaemic by intensified insulin treatment. Treatment of diabetic animals with aminoguanidine or NN0028 prevented the depletion in the nNOS-containing neuron number, an effect not reproduced by L NAME. These studies suggest that the action of AG in restoring the number of nNOS containing retinal neurons is mediated by the inhibition of AGE formation. The depletion of nNOS-containing neurons may contribute to alterations in the autoregulation of blood flow which occurs in diabetes. PMID- 9867209 TI - Increased expression of endothelial cell nitric oxide synthase (ecNOS) in afferent and glomerular endothelial cells is involved in glomerular hyperfiltration of diabetic nephropathy. AB - The overproduction of nitric oxide (NO) is reported in the diabetic kidney and considered to be involved in glomerular hyperfiltration. The precise mechanism of NO production in the diabetic kidney is, however, not known. In this report, we compare the localization of endothelial cell nitric oxide synthase (ecNOS) isoform expression in the kidney tissue of streptozotocin (STZ)-induced diabetic rats and 5/6 nephrectomized rats and clarify the pivotal role of ecNOS for the glomerular hyperfiltration in the early stages of diabetic nephropathy. In diabetic rats, the diameters of afferent arterioles, the glomerular volume, creatinine clearance, and urinary NO2/NO3 were increased after the induction of diabetes. Efferent arterioles were, however, not altered. Insulin or L-NAME treatment returned the diameters of afferent arterioles, glomerular volume, creatinine clearance, and urinary NO2/NO3 to normal. The expression of ecNOS in afferent arterioles and glomeruli of diabetic rats increased during the early stages of the disease, but was not altered in efferent arterioles. Treatment with either insulin or L-NAME decreased ecNOS expression in afferent arterioles and in glomeruli. In contrast, the ecNOS expression was upregulated in both afferent and efferent arterioles and in the glomeruli of 5/6 nephrectomized rats, where the dilatation of afferent and efferent arterioles and glomerular enlargement were observed. Treatment with L-NAME ameliorated the ecNOS expression and dilatation of arterioles. We conclude that enhanced NO synthesis by ecNOS in afferent arterioles and glomerular endothelial cells in response to the hyperglycaemic state could cause preferential dilatation of afferent arterioles, which ultimately induces glomerular enlargement and glomerular hyperfiltration. PMID- 9867210 TI - Advanced glycation endproducts inhibit prostacyclin production and induce plasminogen activator inhibitor-1 in human microvascular endothelial cells. AB - Several thrombogenic abnormalities are associated with diabetes. To investigate the underlying molecular mechanisms, we examined the effects of advanced glycation endproducts (AGE), non-enzymatically glycated protein derivatives, on the production of prostacyclin (PGI2), an anti-thrombogenic prostanoid, and of plasminogen activator inhibitor-1 (PAI-1), a fast-acting serine protease inhibitor of fibrinolysis, in human microvascular endothelial cells (EC). Firstly, AGE-bovine serum albumin (BSA) but not non-glycated BSA, was found to considerably decrease the production of PGI2 to about two-thirds of the control value. Secondly, quantitative reverse transcription-polymerase chain reaction showed that AGE-BSA increased the EC levels of mRNA coding for PAI-1, this being associated with a concomitant increase in the immunoreactive PAI-1 contents and the anti-fibrinolytic activity. Thirdly, the effects of AGE on PGI2 and PAI-1 syntheses in EC were found to be mediated by a receptor for AGE (RAGE) because antisense DNA against RAGE mRNA could reverse the AGE effects. Further, it was found that AGE decreased the intracellular cyclic AMP concentrations in EC and that cyclic AMP agonists such as dibutyryl cyclic AMP, forskolin and PGI2 analogue reduced the AGE-stimulated PAI-1 production, suggesting the involvement of cyclic AMP in the AGE-signalling pathway. The results thus suggest that AGE have the ability to cause platelet aggregation and fibrin stabilization, resulting in a predisposition to thrombogenesis and thereby contributing to the development and progression of diabetic vascular complications. PMID- 9867211 TI - Diabetes-induced changes in lens antioxidant status, glucose utilization and energy metabolism: effect of DL-alpha-lipoic acid. AB - The study was aimed at evaluating changes in lens antioxidant status, glucose utilization, redox state of free cytosolic NAD(P)-couples and adenine nucleotides in rats with 6-week streptozotocin-induced diabetes, and to assess a possibility of preventing them by DL-alpha-lipoic acid. Rats were divided into control and diabetic groups treated with and without DL-alpha-lipoic acid (100 mg x kg body weight(-1) x day(-1), i.p.). The concentrations of glucose, sorbitol, fructose, myo-inositol, oxidized glutathione, glycolytic intermediates, malate, alpha glycerophosphate, and adenine nucleotides were assayed in individual lenses spectrofluorometrically by enzymatic methods, reduced glutathione and ascorbate- colorimetrically, and taurine by HPLC. Free cytosolic NAD+:NADH and NADP+:NADPH ratios were calculated from the lactate dehydrogenase and malic enzyme systems. Sorbitol pathway metabolites were found to increase, and antioxidant concentrations were reduced in diabetic rats compared with controls. The profile of glycolytic intermediates (increase in glucose 6-phosphate and fructose 6 phosphate, decrease in fructosel,6-diphosphate, increase in dihydroxyacetone phosphate, 3-phosphoglycerate, phosphoenolpyruvate, pyruvate, and no change in lactate), and 5.9-fold increase in alpha-glycerophosphate suggest diabetes induced inhibition of glycolysis. Free cytosolic NAD+:NADH ratios, ATP levels, ATP/ADP x inorganic phosphate (Pi), and adenylate charge were reduced in diabetic rats while free cytosolic NADP+:NADPH ratios were elevated. Diabetes-induced changes in the concentrations of antioxidants, key glycolytic intermediates, free cytosolic NAD+:NADH ratios, and energy status were partially prevented by DL alpha-lipoic acid, while sorbitol pathway metabolites and free cytosolic NADP+:NADPH ratios remained unaffected. In conclusion, diabetes-induced impairment of lens antioxidative defense, glucose intermediary metabolism via glycolysis, energy status and redox changes are partially prevented by DL-alpha lipoic acid. The findings support the important role of oxidative stress in lens metabolic imbalances in diabetes. PMID- 9867212 TI - cAMP regulates nitric oxide production and ouabain sensitive Na+, K+-ATPase activity in SH-SY5Y human neuroblastoma cells. AB - We investigated the relation between cyclic AMP (cAMP) and nitric oxide (NO) production, as well as the effect of NO on Na , K+-ATPase activity in the human neuroblastoma cell line SH-SY5Y. Two cAMP agonists, dibutyryl cAMP (DBC) and beraprost sodium (BPS), increased cAMP accumulation and NO production in a time and dose dependent manner at 50 mmol/l glucose. On the other hand, cellular sorbitol and myo-inositol contents and protein kinase C activity were not altered by DBC or BPS. A specific protein kinase A inhibitor, H-89, suppressed increases in nitrite/nitrate and cyclic GMP (cGMP) and protein kinase A activity stimulated by DBC or BPS. This finding suggests that cAMP stimulates NO production by activating protein kinase A via a pathway different from the sorbitol-myo inositol-protein kinase C pathway. We observed that an NO donor, sodium nitroprusside, and an NO agonist, L-arginine, enhanced ouabain sensitive Na+, K+ ATPase activity at 50 mmol/l glucose. We also found that a nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), inhibited Na+, K+-ATPase activity at 5 mmol/l glucose, and partially suppressed the enzyme activity stimulated by DBC or BPS. The results of this study suggest that cAMP regulates protein kinase A activity, NO production and ouabain sensitive Na+, K+-ATPase activity in a cascade fashion. The results also suggest that protein kinase A at least partially regulates Na+, K+-ATPase activity without mediation by NO in SH SY5Y cells. We speculate that cAMP and NO are two important regulatory factors in the pathogenesis of diabetic neuropathy. PMID- 9867213 TI - Increased intestinal glucose absorption and postprandial hyperglycaemia at the early step of glucose intolerance in Otsuka Long-Evans Tokushima Fatty rats. AB - Otsuka Long-Evans Tokushima Fatty (OLETF) rats are reported to be obese Type II (non-insulin-dependent) diabetic rats with insulin resistance and impaired insulin secretion. To investigate the contribution of intestinal glucose absorption to postprandial hyperglycaemia, we determined the plasma xylose concentrations after an 0.8 g/kg oral xylose load which was used as a test of small intestinal glucose absorption in 6-week-old OLETF rats and weight-matched Long-Evans Tokushima Otsuka (LETO) rats. An oral glucose tolerance test showed that OLETF rats developed hyperglycaemia at 60 and 90 min after the glucose load, though the fasting plasma glucose concentration, insulin concentration and insulin-induced in vivo glucose utilization rate were similar. Consistently, in an oral D-xylose loading test, the peak concentration of plasma xylose in OLETF rats was increased by 58.7% compared with that of LETO rats (p < 0.005). The disappearance rate of plasma xylose concentrations after intravenous xylose loading did not differ between the two strains. Co-treatment with 0.4 g/kg phlorizin, a specific inhibitor of sodium-dependent glucose transporter 1 (SGLT1), abolished both plasma glucose and xylose concentrations after the loads. Morphological studies showed that both the small intestinal wet weight and surface area were 30% larger in the OLETF rats than in the LETO rats. Furthermore, the SGLT1 mRNA content of OLETF rats also increased compared with LETO rats. These results suggest that an increased SGLT1 expression concomitant with intestinal hypertrophy in OLETF rats is partly associated with postprandial hyperglycaemia before the onset of insulin resistance and hyperinsulinaemia. PMID- 9867214 TI - Induction of rat muscle insulin resistance by epinephrine is accompanied by increased interstitial glucose and lactate concentrations. AB - Muscle glucose uptake and lactate release during beta-adrenergic stimulation by epinephrine (epi) and beta-adrenergic blockade by propranolol (prop) were investigated during an euglycaemic hyperinsulinaemic (30 pmol x kg(-1) x min(-1)) with or without added somatostatin (0.1 microg/min; pancreatic) clamp in female rats. To assess the interstitial insulin, glucose and lactate concentrations, microdialysis was done in the medial femoral muscle in both legs. The influence of muscle skeletal blood flow on interstitial insulin, glucose and lactate was examined with the microsphere technique, using 57Co-microspheres. Epinephrine decreased glucose infusion rate by about 75% (p < 0.0001) and increased concentrations of interstitial glucose by about 35% (p < 0.001) and lactate by about 65% (p < 0.01). Plasma insulin concentration increased during beta adrenergic stimulation by about 25% (p < 0.05) whereas the interstitial insulin concentration was unchanged. Muscle blood flow in the hindlimb was considerably enhanced by about 130%, (p < 0.001) by epinephrine. Infusion of propranolol totally abolished all the above effects induced by epinephrine. The data show that insulin resistance and vasodilation induced by beta-adrenergic stimulation with epinephrine is accompanied by increased interstitial glucose as well as lactate concentrations in muscle. The increased interstitial glucose concentration is the result of a decreased cellular uptake of glucose together with an increased capillary delivery of glucose by vasodilation. It is concluded that the severe cellular resistance to insulin induced by epinephrine could not be overcome either by the increased insulin secretion or by vasodilation. PMID- 9867215 TI - A search for tyrosine kinase receptors expressed in the rat embryonic pancreas. AB - It is known that during embryonic life, interactions between the pancreatic epithelium and its surrounding mesenchyme are important for proper development of the pancreas. These interactions are thought to be mediated by soluble factors, which could be, as in other tissues, the ligands of tyrosine kinase receptors. In this study, we screened for tyrosine kinase receptors expressed in pancreata of 13-day-old embryonic rats. Using a polymerase chain reaction-based approach that exploits sequence similarities within the catalytic kinase domains of these receptors, we identified 30 different tyrosine kinase receptors. The same approach was then used on cDNA prepared from fractions enriched in epithelium or in mesenchyme. Receptors for factors such as platelet derived growth factors were found to be expressed both in the epithelial and the mesenchymal fractions. Receptors for stem cell factor, for epidermal growth factor family members were mainly found in the epithelial fraction. The profile of expression of receptor tyrosine kinases in the embryonic pancreas was finally compared to the one found in other tissues and cell types, such as kidney, brain or INS-1 cells. Platelet derived growth factor receptors and ErbB2 were found to be enriched in the embryonic pancreas when compared with other tissues. It will now be possible to test the effects of the ligands of the different receptors we have cloned, on the differentiation and growth of the pancreas. PMID- 9867216 TI - Inhibition of glucose stimulated insulin secretion by neuropeptide Y is mediated via the Y1 receptor and inhibition of adenylyl cyclase in RIN 5AH rat insulinoma cells. AB - Neuropeptide Y (NPY) has been shown to inhibit insulin secretion from the islets of Langerhans. We show that insulin secretion in the insulinoma cell line RIN 5AH is inhibited by NPY. 125I-Peptide YY (PYY) saturation and competition-binding studies using NPY fragments and analogues on membranes prepared from this cell line show the presence of a single class of NPY receptor with a Y1 receptor subtype-like profile. Inhibition of insulin secretion in this cell line by NPY fragments and analogues also shows a Y1 receptor-like profile. Both receptor binding and inhibition of insulin secretion showed the same orders of potency with NPY > [Pro34]-NPY > NPY 3-36 >> NPY 13-36. The Y1 receptor antagonist, BIBP 3226, blocks NPY inhibition of insulin secretion from, and inhibits 125I-PYY binding to, RIN 5AH cells. Northern blot analysis using a Y1-receptor specific probe shows that NPY Y1 receptors are expressed by RIN 5AH cells. Y5 receptors are not expressed in this cell line. Neuropeptide Y inhibition of insulin secretion is blocked by incubation with pertussis toxin, implying that the effect is via a G-protein (Gi or Go) coupled receptor. Neuropeptide Y inhibits the activation of adenylyl cyclase by isoprenaline in RIN 5AH cell lysates, and the stimulation of cAMP by glucagon-like peptide-1 (7-36) amide (GLP-1). It also blocks insulin secretion stimulated by GLP-1, but not by dibutyryl cyclic AMP. Hence, we suggest that NPY inhibits insulin secretion from RIN 5AH cells via a Y1 receptor linked through Gi to the inhibition of adenylyl cyclase. PMID- 9867217 TI - Cellular characterization of pituitary adenoma cell line (AtT20 cell) transfected with insulin, glucose transporter type 2 (GLUT2) and glucokinase genes: insulin secretion in response to physiological concentrations of glucose. AB - We investigated the mechanisms of insulin secretion by transfecting into a pituitary adenoma cell line (AtT20) a combination of genes encoding human insulin (HI), glucose transporter type 2 (GLUT2) and glucokinase (GK), followed by studying the characteristics of these cells. In static incubation, a cell line transfected with insulin gene alone (AtT20HI) secreted mature human insulin but this was not in a glucose-dependent manner. Other cell lines transfected with insulin and GLUT2 genes (AtT20HI-GLUT2-3) or with insulin and GK genes (AtT20HI GK-1) secreted insulin in response to glucose concentrations of only less than 1 mmol/l. In contrast, cell lines transfected with insulin, GLUT2 and GK genes (AtT20HI-GLUT2-GK-6, AtT20HI-GLUT2-GK-7 and AtT20HI-GLUT2-GK-10) showed a glucose dependent insulin secretion up to 25 mmol/l glucose. Glucose utilization and oxidation were increased in AtT20HI-GLUT2-GK cell lines but not in AtT20HI, AtT20HI-GLUT2-3 and AtT20HI-GK-1 cells at physiological glucose concentrations, compared with AtT20 cells. Diazoxide, nifedipine and 2-deoxy glucose suppressed (p < 0.05) glucose stimulated insulin secretion in AtT20HI-GLUT2-GK-6 cells. Glibenclamide, KCl or corticotropin releasing factor (CRF) stimulated (p < 0.05) insulin secretion both in AtT20HI and AtT20HI-GLUT2-GK-6 cells. Insulin secretion stimulated by glibenclamide, KCl or CRF was further enhanced by the addition of 25 mmol/l glucose in AtT20HI-GLUT2-GK-6 cells but not in AtT20HI cells. In perifusion experiments, a stepwise increase in glucose concentration from 5 to 25 mmol/l stimulated insulin secretion in AtT20HI-GLUT2-GK cell lines but the response lacked a clear first phase of insulin secretion. Our results suggest that both GLUT2 and glucokinase are necessary for the glucose stimulated insulin secretion in at least rodent cell lines, and that other element(s) are necessary for a biphasic insulin secretion typically observed in beta cells. PMID- 9867218 TI - Lymphoid hyperplasia, CD45RBhigh to CD45RBlow T-cell imbalance, and suppression of Type I diabetes mellitus result from TNF blockade in NOD-->NOD-scid adoptive T cell transfer. AB - Sustained antibody-mediated inhibition of tumor necrosis factor (TNF) activity offers protection against Type I (insulin-dependent) diabetes mellitus in non obese diabetic (NOD) mice. The mechanism of this effect, however, has remained obscure: TNFalpha might be required for the development of specific immune responses to islet antigens or it could directly participate in destruction of beta cells. In this study, autoimmune destruction of beta cells was initiated in NOD-severe combined immunodeficient (scid) mice by transfer of NOD splenic T cells to induce diabetes. The blockade of TNFalpha activity was achieved during a narrow window of time after transfer. Transient inhibition of TNFalpha greatly reduced the number of islet lymphocytes and the incidence of diabetes in recipients of prediabetic NOD spleen cells. Protection extended beyond the interval of effective TNF blockade. Furthermore, the protective effect was only observed if cells were obtained from 6-week-old donors. The suppression of autoimmunity was reversible in the context of adoptive transfer as indicated by the transfer of splenocytes from the primary recipient to a second NOD-scid host led to a diabetic outcome. The blockade of TNFalpha was accompanied by a considerable increase in spleen size and doubling of the total splenocyte count, suggesting that TNFalpha might normally eliminate a transplanted T-cell subset within the recipients. Further analysis showed an increase in the absolute count of CD4 + T cells and pronounced distortion of the CD45RBhigh to CD45RBlow ratio, with a relative augmentation in the CD45RBlow count in the spleen. TNFalpha appears to regulate the number and subtype distribution of a transplanted T cell population. PMID- 9867219 TI - Missense mutations in the pancreatic islet beta cell inwardly rectifying K+ channel gene (KIR6.2/BIR): a meta-analysis suggests a role in the polygenic basis of Type II diabetes mellitus in Caucasians. AB - The K+ inwardly rectifier channel (KIR) is one of the two sub-units of the pancreatic islet ATP-sensitive potassium channel complex (I(KATP)), which has a key role in glucose-stimulated insulin secretion and thus is a potential candidate for a genetic defect in Type II (non-insulin-dependent) diabetes mellitus. We did a molecular screening of the KIR6.2 gene by single strand conformational polymorphism (SSCP) and direct sequencing in 72 French Caucasian Type II diabetic families. We identified three nucleotide substitutions resulting in three amino acid changes (E23K, L270V and 1337V), that have also been identified in other Caucasian Type II diabetic subjects. These variants were genotyped in French cohorts of 191 unrelated Type II diabetic probands and 119 normoglycaemic control subjects and association studies were done. The genotype frequencies of the L270V and 1337V variants were not very different between Type II diabetic subjects and control groups. In contrast, analysis of the E23K variant showed that the KK homozygocity was more frequent in Type II diabetic than in control subjects (27 vs 14%, p = 0.015). Analyses in a recessive model (KK vs EK/EE) tended to show a stronger association of the K allele with diabetes (p = 0.0097, corrected p-value for multiple testing < 0.02). The data for the E23K variant obtained here and those obtained from three other Caucasian groups studied so far were combined and investigated by meta-analysis. Overall, the E23K variant was found to be significantly associated with Type II diabetes (0.001 < or = p < or = .00106, corrected p-values for multiple testing p < or = 0.01). This study shows that KIR6.2 polymorphisms are frequently associated with Type II diabetes in French Caucasians. Furthermore, a meta-analysis combining different Caucasian groups suggests an significant role of KIR6.2 in the polygenic context of Type II diabetes. PMID- 9867220 TI - The putative role of the hormone-sensitive lipase gene in the pathogenesis of Type II diabetes mellitus and abdominal obesity. AB - Impaired lipolysis has been proposed as a pathogenic factor contributing to clustering of abdominal obesity and dyslipidaemia in Type II (non-insulin dependent) diabetes mellitus--that is, the metabolic syndrome (MSDR). As this syndrome clusters in families, alterations in the hormone-sensitive lipase (HSL) gene could contribute to the genetic predisposition to MSDR. To test this hypothesis we carried out population and intrafamily association studies in individuals with MSDR, using a polymorphic marker (LIPE) in the HSL gene. There was a significant difference in allele frequency distribution between 235 Type II diabetic patients and 146 control subjects (p = 0.002), particularly between 78 abdominally obese Type II diabetic patients with MSDR and the control group (p = 0.010). An extended transmission disequilibrium test (TDT) showed transmission disequilibrium of 66 alleles to 42 nondiabetic, abdominally obese offspring in families with Type II diabetes (p < 0.05). A slight difference in allele frequency distribution was seen between 71 individuals from the lowest and 71 from the highest tertile of isoprenaline-induced lipolysis in fat tissue (p = 0.07). No missense mutations were found with single-strand conformational polymorphism (SSCP) in 20 abdominally obese subjects with MSDR. In conclusion, our population and intrafamily association studies suggest that the LIPE marker in the HSL gene is in linkage disequilibrium with an allele and/or gene which increases susceptibility to abdominal obesity and thereby possibly to Type II diabetes. PMID- 9867221 TI - Insulin inhibits inducible nitric oxide synthase in skeletal muscle cells. AB - Recent studies have shown that cytokines and endotoxins impair insulin-stimulated glucose transport by activating the expression of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) production in skeletal muscle cells. In this study, we investigated whether iNOS induction is modulated by insulin in L6 myocytes. Long term exposure of muscle cells to tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS) greatly increased iNOS mRNA expression and NO production. Addition of insulin to the cytokine/LPS treated muscle cells reduced (by approximately 40%) NO production. This inhibition was similar to that observed with the synthetic glucocorticoid dexamethasone, a known inhibitor of iNOS in several cell types. The combination of insulin and dexamethasone was more effective than either agent alone in reducing NO production. Dexamethasone greatly inhibited the effect of cytokines/LPS to induce cellular iNOS mRNA expression. In strong contrast, insulin failed to reduce iNOS mRNA expression under similar conditions. These results show that insulin is a novel inhibitor of iNOS-mediated NO production in skeletal muscle cells. Furthermore, our data indicate that unlike glucocorticoids, insulin does not inhibit NO production by suppression of iNOS gene transcription. PMID- 9867222 TI - Mutations in the hepatocyte nuclear factor-1alpha gene in Caucasian families originally classified as having Type I diabetes. AB - Mutations in the hepatocyte nuclear factor-1alpha (HNF-1alpha) gene are the cause of maturity-onset diabetes of the young type 3 (MODY3), which is characterised by a severe impairment of insulin secretion and an early onset of the disease. Also at onset of diabetes some MODY patients show similar clinical symptoms and signs as patients with Type I (insulin-dependent) diabetes mellitus. The objective of this study was to estimate the prevalence of MODY3 patients misclassified as Type I diabetic patients. From a large population-based sample of unrelated Danish Caucasian Type I diabetic patients with an affected first degree relative, 39 patients (6.7%) who did not carry any high-risk HLA-haplotypes, i.e. DR3 or DR4 or both were examined by single-strand conformational polymorphism scanning and direct sequencing of the coding region and the minimal promoter of the HNF-1alpha gene. Four of the 39 Type I diabetic patients (10%) were identified as carrying mutations in the HNF-1alpha gene. One patient carried a missense mutation (Glu48Lys) in exon 1, two patients carried a missense mutation (Cys241Gly) in exon 4 and one patient carried a frameshift mutation (Pro291fsdelA) in exon 4. The mutations were all identified in heterozygous form, segregated with diabetes, and were not identified in 84 unrelated, healthy subjects. Furthermore, family history in three of the four families showed diabetes in four consecutive generations, suggestive of an autosomal dominant inheritance. In conclusion, about 10% of Danish diabetic patients without a high-risk HLA-haplotype, originally classified as having Type I diabetes could have diabetes caused by mutations in the HNF-1alpha gene. Clinical awareness of family history of diabetes and mode of inheritance might help to identify and reclassify these diabetic subjects as MODY3 patients. PMID- 9867224 TI - Trp64Arg mutation of beta3-adrenoceptor gene is associated with diabetic nephropathy in Type II diabetes mellitus. PMID- 9867223 TI - Islet autoimmunity before and after pancreas transplantation in patients with Type I diabetes mellitus. PMID- 9867225 TI - Antibodies directed against the ectodomain of ICA512: is there any evidence in patients with diabetes mellitus? PMID- 9867226 TI - Role of acoustic striae in hearing: reflexive responses to elevated sound sources. AB - This report is the fourth in a series describing the results of ablation-behavior experiments directed to the ascending output of the cochlear nuclei as it is conducted centrally within the acoustic striae. This fourth report focuses on the unique physiology of the fusiform or 'output' cells of the dorsal cochlear nucleus whose axons course through the dorsal acoustic stria (DAS). Because electrophysiological studies have shown that the cues for sensing the elevation of a sound source would seem to be best analyzed by the dorsal cochlear nucleus and projected centrally via its DAS, we tested normal cats and cats deprived of DAS for their ability to orient to elevated sources of broad-band noise. For behavioral testing, we made use of reflexive or unconditioned orienting responses to elevated sound sources using a similar method to one we have used previously for azimuth testing (Thompson GC, Masterton RB. Brainstem auditory pathways involved in reflexive head orientation to sound. J Neurophysiol 1978;41:1183 1202). The results show that cats deprived of their DAS do indeed have a marked deficit in their ability to orient to an elevated sound source. Further behavioral testing indicated that this deficit is not the secondary result of an attentional or peripheral motor deficit. Although the present results do not prove that the reflexive deficit is strictly auditory in nature, the deficit is notable in that it is the only one yet known to result from a lesion of the dorsal cochlear nucleus or its central projections. PMID- 9867227 TI - Differential alterations in basal and D-amphetamine-induced behavioural pattern following 6-OHDA or ibotenic acid lesions into the dorsal striatum. AB - It is well known that the corpus striatum is related to the sterotyped activation induced by several psychostimulants. In this study we analyzed the effects of 6 OHDA, in comparison with those of ibotenic acid lesions, into the dorsal striatum, on the behavioural pattern induced by saline or D-amphetamine treatment. A computerized technique for recording the animal motor activity was developed in order to define in a detailed way the behavioural profile in lesioned and sham-operated rats induced by the saline or D-amphetamine treatment. A 6-OHDA lesion into the dorsal striatum modified the basal behavioural pattern which was mainly characterized by reduced motor activation while ibotenic acid lesion affected the structure of the basal behavioral pattern. D-Amphetamine administration in 6-OHDA lesioned rats induced a behavioural stimulation, but a decreased motor and stereotyped activation was observed compared to the sham operated animals treated with D-amphetamine. In contrast, D-amphetamine administration in the ibotenic acid-lesioned rats induced a motor and stereotyped activity which was not reduced compared to that seen after D-amphetamine treatment in sham-operated rats. These results suggest that these two types of lesion induced differential effects on the behavioural pattern either after saline or after D-amphetamine administration. Dopaminergic neurotransmission in the dorsal striatum plays a permissive role on the emergence of the behavioural responses, while the dorsal striatum circuitry plays a crucial role on the organization of the behavioural pattern. In addition, dopaminergic activity in this structure serves a primary control in the D-amphetamine-elicited motor activation or stereotypy, while the striatal structure is involved in the shaping of the D-amphetamine behavioural pattern. PMID- 9867228 TI - Effects of MK-801 on vicarious trial-and-error and reversal of olfactory discrimination learning in weanling rats. AB - The effects of dizocilpine maleate (MK-801) on vicarious trial-and-error (VTE), and on simultaneous olfactory discrimination learning and its reversal, were observed in weanling rats. The term VTE was used by Tolman (The determiners of behavior at a choice point. Psychol. Rev. 1938;46:318-336), who described it as conflict-like behavior at a choice-point in simultaneous discrimination learning. It takes the form of head movements from one stimulus to the other, and has recently been proposed by Amsel (Hippocampal function in the rat: cognitive mapping or vicarious trial-and-error? Hippocampus, 1993;3:251-256) as related to hippocampal, nonspatial function during this learning. Weanling male rats received systemic MK-801 either 30 min before the onset of olfactory discrimination training and its reversal, or only before its reversal. The MK-801 treated animals needed significantly more sessions to acquire the discrimination and showed significantly fewer VTEs in the acquisition phase of learning. Impaired reversal learning was shown only when MK-801 was administered during the reversal-learning phase, itself, and not when it was administered throughout both phases. PMID- 9867229 TI - Repeated injection of GBR 12909, but not cocaine or WIN 35,065-2, into the ventral tegmental area induces behavioral sensitization. AB - A role for the mesolimbic dopamine system in the development of behavioral sensitization to psychostimulants, such as cocaine and amphetamine, is well established. Previous reports have suggested that the ventral tegmental area (VTA) is involved in the initiation of, while the nucleus accumbens is in involved in the expression of behavioral sensitization. This hypothesis is supported in part, by studies which demonstrated that behavioral sensitization could be induced by repeated intra-VTA, but not intra-accumbal, administration of amphetamine. The present studies were designed to determine whether repeated intra-VTA cocaine would similarly induce behavioral sensitization. Rats receiving four daily injections of cocaine (1.5, 5 or 15 nmol/side) into the VTA did not show a sensitized behavioral response when challenged with cocaine (15 mg/kg, ip) 1 week later. In contrast to this, repeated injection of the specific dopamine reuptake inhibitor, GBR 12909 (15 nmol/side) produced behavioral sensitization to a challenge injection of cocaine. Repeated injections of the cocaine analog WIN 35,065-2 did not induce behavioral sensitization to cocaine, suggesting that the local anesthetic properties of cocaine were not responsible for the inability of intra-VTA cocaine to induce sensitization. In summary, the data suggest that sensitization to cocaine may involve mechanisms different from amphetamine. PMID- 9867230 TI - Lesions of the midline thalamic nuclei impair classical conditioning under partial but not continuous reinforcement conditions. AB - Rabbits with lesions of the midline thalamic nuclei were compared to rabbits with sham lesions on classical eyeblink (EB) and heart rate (HR) conditioning. Separate groups of sham and lesioned animals received either 25, 50 or 100% reinforcement with a periorbital shock unconditioned stimulus. Animals with lesions showed slightly impaired EB conditioning, compared to sham animals, under the partial but not continuous reinforcement schedules. Midline lesions also reduced the magnitude of the decelerative HR conditioned response under partial but not continuous reinforcement conditions. These findings suggest that the midline thalamic nuclei, like the mediodorsal nucleus of the thalamus, process information required for response selection under non-optimal learning conditions. PMID- 9867231 TI - Simulation of psychosis by continuous delivery of phencyclidine from controlled release polymer implants. AB - To simulate psychosis in rats we have developed a method for the continuous delivery of phencyclidine (PCP) using implantable controlled-release polymers. PCP polymer implants produced deficits in latent inhibition which do not occur after repeated bolus injections. PCP implanted rats were also devoid of any anxiogenic signs, motoric hyperactivity and learning acquisition which can be seen in rats receiving daily bolus injections of a comparable PCP dose. This behavioral double-dissociation of the two modes of PCP application was accompanied by respective neurochemical changes. PCP binding sites were reduced in both striatum and hippocampus, but in the hippocampus, loss of PCP binding sites was more severe following pulsatile PCP administration. Morphological assessment revealed a significant shrinkage of the CA3 region in hippocampus in both groups. Pharmacokinetic analysis showed that the maximum PCP concentration in the brain after bolus injections was 10-fold above the PCP implants. PMID- 9867232 TI - Encoding and retrieval in declarative learning: a positron emission tomography study. AB - We present neuroanatomical correlates of encoding and retrieval in an episodic memory task using visually presented highly imaginable word-pair associates. A total of 13 right-handed normal male volunteers took part in the study. Each subject underwent six (15)O-butanol PET scans. On each of the six trials the memory task began 30 s before the injection of a bolus of (15)O-butanol. The subjects had to learn and retrieve 12 word pairs (highly imaginable words, not semantically related, hard associations). The presentation of nonsense words served as a reference condition. Recall accuracy after 2-4 presentations was 66.1%+/-21.1 correct during the PET measurement so that scanning during the retrieval of word pair associates was appropriate to capture the brain activity associated with retrieval. The results obtained support the hypothesis of the presence of an asymmetric network consisting of distributed brain structures subserving associative memory. We show left dorsolateral prefrontal activation during the encoding of visually presented word pair associates, whereas retrieval led to bilateral frontal activation. Furthermore, the importance of the precuneus in the retrieval of highly imaginable word-pair associates using visual imagery as a mnemonic strategy is demonstrated. PMID- 9867233 TI - Muscimol suppression of the dorsal cochlear nucleus impairs frequency discrimination in rats. AB - The cochlear nucleus is composed of three sub-nuclei: the dorsal (DCN), anteroventral (AVCN) and posteroventral cochlear nucleus (PVCN). In addition to connections between these sub-nuclei, each nucleus receives frequency specific tonotopically organised input from the cochlea. Evidence suggests that connections from the DCN to the AVCN are inhibitory and organised tonotopically but the functional significance of this pathway has yet to be elucidated. The possible role of this pathway in frequency discrimination using a T-maze behavioural paradigm and DCN suppression was examined. Five rats were trained on a two choice frequency discrimination task. Once frequency difference limens for 10-30% performance above chance were determined, rats had cannulae implanted bilaterally over the DCN. After recovery rats were tested on the behavioural task with nothing, saline and the GABA agonist muscimol injected into the DCN via the cannulae. Muscimol alone significantly reduced the rats ability to perform the task. This performance decrease was attributed to an inability to discriminate high frequency and not low frequency tones suggesting that place and not temporal coding of sound was compromised by DCN suppression. These results are consistent with the hypothesis that inhibitory drive from the DCN to AVCN may be crucial for the fine tuning of frequency information. PMID- 9867234 TI - Relationship between dendritic pruning and behavioral recovery following sensorimotor cortex lesions. AB - A unilateral injury to the forelimb area of the sensorimotor cortex results in an increase in dendritic arborization in the contralateral homotopic cortex which is followed by a pruning back of these dendritic arbors. The increase in arborization is due to an increase in the use of the unimpaired forelimb for postural-motor support; whereas, the dendritic pruning is related, in time, to the return to more symmetrical limb use, but is not prevented by the maintenance of asymmetrical limb use. Dendritic pruning can be prevented by administering an NMDA receptor antagonist (such as MK801 or ethanol) during the pruning phase. This manipulation also coincides with the chronic reinstatement of behavioral deficits. The purpose of this study was to see whether removing the antagonism of the NMDA receptor results in the eventual return of dendritic pruning and behavioral recovery. Therefore, MK801 was administered to lesioned animals starting at post-lesion day 18. One group received MK801 injections until day 60 (Lesion + MK60) and another lesioned group received MK801 until day 30 after which the injections were changed to saline until day 60 (Lesion + MK30). Lesion + MK60 animals showed a prevention of dendritic pruning as well as a chronic reinstatement of forelimb deficits. Lesion + MK30 animals also showed a prevention of dendritic pruning, however, they showed behavioral recovery. These findings suggest that pruning of dendritic arbors may not be directly related to behavioral recovery. PMID- 9867235 TI - Progesterone facilitates the acquisition of avoidance learning and protects against subcortical neuronal death following prefrontal cortex ablation in the rat. AB - Following a cortical injury, neurons in areas near and connected to the site of injury begin to degenerate. The observed neuronal death may contribute to the severity of the observed behavioral impairments. The purpose of the present study was to examine if progesterone, a hormone known for its effectiveness at reducing cerebral edema, could protect against secondary neuronal death and facilitate the acquisition of an avoidance learning task in an ablation model of cortical injury. Rats served as sham controls or received bilateral ablation of the medial prefrontal cortex followed by a 10-day regimen of progesterone (4 mg/kg) or oil vehicle (1 ml/kg) beginning 1 h after cortical lesions. Progesterone-treated lesion rats showed a significant facilitation of avoidance learning compared to oil-treated lesion controls. In addition, progesterone-treated lesion animals did not differ from either progesterone- or oil-treated sham controls in avoidance learning. Anatomical analysis revealed that progesterone treatment decreased the amount of neuronal death seen in the striatum and the mediodorsal nucleus of the thalamus. The findings are consistent with the notion that progesterone is an effective neuroprotective agent and suggest that the hormone can reduce the behavioral impairments associated with frontal cortical ablation injury. PMID- 9867236 TI - The medial frontal cortex and temporal memory: tests using spontaneous exploratory behaviour in the rat. AB - Evidence from several sources indicates that the frontal cortex is involved in memory involving time, or temporal memory, and in behavioral sequencing. Rats were presented with two familiar objects and the amount of exploration directed at each was recorded. Subjects directed more exploration at the object encountered further ago in the past, even though the two objects differed by only 1 h. After bilateral medial frontal cortex lesion, animals did not differentiate between old and recent objects. Performance on an object recognition test (novel versus familiar objects) that also used exploration time did not change as a result of the medial frontal cortex lesions. PMID- 9867237 TI - Discrete quinolinic acid lesions of the rat prelimbic medial prefrontal cortex affect cocaine- and MK-801-, but not morphine- and amphetamine-induced reward and psychomotor activation as measured with the place preference conditioning paradigm. AB - As a part of the mesocorticolimbic system, the medial prefrontal cortex (mPFC) is thought to participate in the regulation of locomotor activity, motivation and reward. The mPFC consists of at least three different subareas. In previous lesion studies examining this region usually large parts of the mPFC were destroyed, with little discrimination between the different subareas. Therefore, this study was designed to selectively lesion the prelimbic area of the mPFC using a relatively low dose of quinolinic acid. In a conditioned place preference (CPP) experiment, lesioned and control animals were treated with cocaine (15 mg/kg), amphetamine (4 mg/kg), morphine (10 mg/kg) or MK-801 (0.3 mg/kg) to induce CPP. The lesion blocked the development of CPP only in animals receiving cocaine, but not in animals receiving amphetamine or morphine. MK-801 failed to produce a CPP in both lesioned and unlesioned animals. During the conditioning experiment, the acute locomotor response to the different drugs was also measured. Only the response (in terms of locomotion and rearing) to cocaine and MK-801 was reduced to a significant extent by the lesion, while the response to amphetamine and morphine was not affected. Also, the lesions did not cause any changes in the spontaneous activity of the animals when tested without drug. These results show that even small lesions of the prelimbic subarea of the mPFC are sufficient to produce behavioral effects. However, these are apparent only when the animals are challenged with cocaine or MK-801, but not with amphetamine or morphine, or when drug-free. This suggests that the mPFC might have a special role in mediating cocaine and MK-801 effects. PMID- 9867238 TI - An examination of the spatial working memory deficit following neurotoxic medial dorsal thalamic lesions in rats. AB - The present study examined the performance of rats with neurotoxic lesions centred in the thalamic nucleus medialis dorsalis on standard and modified versions of the eight arm radial maze test. In Experiment 1, the thalamic lesions produced a borderline deficit in acquisition of the standard task, but subsequently had no effect when a delay was interposed after the first four arms had been entered. The same lesions had no effect on T-maze alternation, but they did impair radial-arm maze performance when intramaze and extramaze cues were set against each other. In Experiment 2, lesions of the dorsomedial thalamus impaired acquisition of the standard radial-arm maze task, but combining the results from Experiments 1 and 2 showed that this acquisition deficit was confined to those animals in which bilateral damage extended into the adjacent anterior thalamic nuclei. In addition, lesions of the dorsomedial thalamus disrupted radial-arm maze performance when the task was modified to compare working memory and reference memory and increased activity and exploration. These changes were not associated with anterior thalamic damage. Finally, the thalamic lesions did not affect performance on a test of spontaneous object recognition. It is concluded that lesions of medialis dorsalis do not disrupt spatial memory but do affect other processes that can interact with task performance. These include a failure of extramaze cues to overshadow intramaze cues, a change in activity and exploration levels and deficits in with-holding spatial responses. PMID- 9867239 TI - Sexual behavior and wet dog shakes in the male rat: regulation by corticosterone. AB - The potential involvement of adrenal steroids in the regulation of 'wet dog shakes' (WDS) and sexual behavior was investigated in male rats treated or not with the serotonergic type 2A (5-HT2A) agonist DOI (5-HT2A receptor agonist 1 (2,5-dimethoxy-4-iodophenyl)-2-aminopropane). In Experiment 1, the frequency of both spontaneous and DOI-induced WDS were compared in adrenalectomized and sham adrenalectomized rats. Adrenalectomy significantly reduced the frequency of DOI induced WDS. In Experiment 2, adrenalectomized and sham-adrenalectomized rats received either corticosterone or oil chronically and were again scored for WDS behavior. Corticosterone effectively blocked the adrenalectomy-induced reduction of WDS in the DOI treatment condition. In Experiment 3, intact male rats were chronically administered either corticosterone or oil treatment. Animals were then compared on measures of both spontaneous and DOI-induced WDS and sexual behavior. Corticosterone significantly increased WDS and inhibited sexual behavior in both the spontaneous and DOI treatment conditions. These results suggest that the adrenal steroid corticosterone is important in the regulation of WDS and sexual behavior in the male rat and that this regulation may be mediated by activity at 5-HT2A receptors. PMID- 9867240 TI - Small platform stress attenuates the anxiogenic effect of diazepam withdrawal in the plus-maze test. AB - The effect of stress on the behavioural changes caused by diazepam withdrawal was studied in mice. Diazepam (2.5 mg/kg) or vehicle was injected intraperitoneally twice a day for 2 weeks. When the last vehicle or diazepam injection had been administered to the mice, 12 h later, the mice were subjected to small platform stress exposure or left in their home cages. Small platform stress was induced by placing mice on small platforms (3.5 cm diameter) surrounded by water for 24 h. This experimental model contains several factors of stress like rapid eye movement (REM) sleep deprivation, isolation, immobilization and falling into the water. The plus-maze test was carried out with stressed mice as well as with mice not subjected to stress. Small platform stress induced an anxiolytic effect and diazepam withdrawal--an anxiogenic effect in the plus-maze test. Small platform stress also attenuated the anxiogenic effect of diazepam withdrawal. On the basis of this data it was proposed that small platform stress counteracts the anxiogenic effect of diazepam withdrawal in the plus-maze test. It was also proposed that the effect of stress on the signs of benzodiazepine withdrawal depends on the characteristics and duration of stress. PMID- 9867241 TI - Neuronal activity of human caudate nucleus and prefrontal cortex in cognitive tasks. AB - Lesions of the caudate nucleus and prefrontal cortex may display similar cognitive deficits. Recent advances in cognitive neuroscience have clarified the functional role of prefrontal cortical areas in certain cognitive operations involved in simple language tasks. We have addressed the role of the caudate nucleus in tasks of lexical decision, semantic categorization, recognition memory, reading aloud and object naming by recording neuronal activity in patients with depth electrodes. During visual processing of words, caudate cells exhibited excitatory responses related to both semantic and phonological articulatory encoding with non-overlapping time courses. The firing rate of the cells was increased when the semantic processing was required. This occurred within 400-600 ms after the stimulus onset, or within the first 200-300 ms of the delay period. The increased firing within 1000-1200 ms after the stimulus onset was related to the phonological processing. These responses turned out to be strikingly similar to those in Broca's area. Both reading aloud and explicit memory retrieval tasks elicited a sustained inhibition of firing of the same cells with a greater onset latency. Chronometric comparison of prefrontal, temporo-parietal and caudate activities in similar tasks relates the time course of these activations to the fronto-caudate anatomical loops and helps further understanding of the anatomy and circuitry involved in human cognition. PMID- 9867242 TI - Differential involvement of the lateral and medial divisions of the septal area on spatial learning processes as revealed by intracranial self-administration of morphine in mice. AB - The present study was conducted to investigate the effects of morphine self administration into either the medial or lateral divisions of the septal region on spatial discrimination abilities in mice. To this end, BALB/c mice received a dose of 5 ng/50 nl of morphine sulfate, via a stainless steel injection cannula, inserted into either the medial septal area (MS) or the lateral septal nucleus (LS), when they are close to the end of one of the two choice arms of a Y-maze (acquisition period). In these conditions a discrimination between the reinforced arm and the neutral arm is acquired in MS as well as in LS mice. However both acquisition and subsequent extinction (vehicle only available) was more rapid in the LS group than in the MS group. Moreover, during the extinction period, numerous escape attempts from the Y-maze were observed for MS mice. When the dose of morphine was raised to 50 ng the pattern of acquisition was unchanged in the LS group. In contrast animals of the MS group learned to avoid the arm where the higher dose of morphine was delivered thus allowing us to conclude that in MS animals the drug effect became aversive at this higher dose. This possibility was directly investigated in a second experiment by closing the access to the neutral arm. Thus, for a 5-ng dose the rewarding effect of morphine was demonstrated in both MS and LS groups by the decrease of self-administration latencies which, furthermore, were notably lower than in the discrimination situation. Moreover, with the dose of 50 ng of morphine the latencies were identical for the two groups and at their lowest value thus indicating that morphine had similar appetitive effects in MS as well as in LS mice in this situation. Thus, the previously observed deficit of MS subjects, including escape attempts, disappeared when the dose of morphine was raised and the experimental context simplified. These results which demonstrate differential functional properties of these two septal divisions are discussed in terms of conflict resulting from the strongly appetitive effects of the morphine which induces, in parallel, deleterious consequences on cognitive processing in MS self-injected mice. PMID- 9867243 TI - Improvement in motor performance of Weaver mutant mice following lesions of the cerebellum. AB - In Weaver mutants (B6CBA wv/wv) cerebellar granule cells degenerate almost completely postnatally. A partial loss of Purkinje cells (PC) and a degeneration of dopaminergic cells in the substantia nigra have also been found. Weaver mice suffer from striking motor symptoms, including difficulty in walking without toppling over. In an attempt to influence the poor motor performance, the cerebellum in young animals was removed, thus eliminating the faulty output of surviving PCs, demonstrated electrophysiologically. Unoperated Weaver, lesioned wildtypes and one sham mouse were used as controls. Before and after operation, a battery of behavioural tests was performed. In Weaver mice, tumbling to the side (t) and the relation of t to the motor activity (k) while traversing an open field matrix, (t/k), improved considerably, as did manoeuvring on a slanted wire mesh, but keeping balance on a wooden bench did not to the same degree. Locomotor activity alone improved in some animals. In wildtypes no significant changes occurred after operation, with the exception of a strong reduction in locomotor activity. The experiments demonstrate that the motor performance in Weaver mutant mice benefits from removal of their cerebellum. PMID- 9867244 TI - Light exposure of the embryo and development of behavioural lateralisation in chicks, I: olfactory responses. AB - Chicks exposed to light during late foetal life experience stimulation of the right eye only and, in consequence, develop asymmetries of the crossed visual projections from thalamus to forebrain and differences in performance of some visual tasks when using the right or left eye. The present study compared dark- and light-incubated chicks in a test of olfaction in which clove oil odour was presented together with a coloured bead. When the chicks were tested with a blue bead and using the right nostril (left nostril occluded by wax), head shaking and pecking were elevated, compared to pretest responses to an unscented, white bead. No significant elevation of head shaking occurred in chicks tested with the blue bead and using the left nostril, although pecking increased, which indicates that these chicks attended to the visual parameters of the bead but not the odour. It appears that, when the left nostril is used, attention to an attractive visual stimulus suppresses responses to olfactory input to the left hemisphere. When the clove oil odour was presented together with a less attractive, red bead, no significant lateralisation emerged. Light or dark experience prior to hatching had no effect on the lateralised performance of the blue-bead test. PMID- 9867245 TI - Prolactin receptor antagonists. AB - Most prolactin (PRL) in the circulation is produced by the pituitary. However, a wide variety of traditional target tissues of the hormone have also been shown to produce their own prolactin. The amount produced per cell is low, but may well be sufficient for autocrine/paracrine activity. Although dopamine agonists allow one to study the target tissue effects of pituitary PRL, other agents, such as PRL receptor (PRLR) antagonists, are needed to analyze autocrine/paracrine loops. With PRLR antagonists, it should be possible to dissect out the role of extrapituitary prolactin in both the normal physiology, and the pathophysiology of various tissues. In tissues where the locally produced PRL may promote disease, such antagonists have the potential to be important therapeutics. This article briefly, but critically, reviews current understanding of PRL-receptor interactions and initial signaling, and describes the development of both growth hormone (GH) and PRL antagonists within that context. In the final section, results with a very potent PRL antagonist further one theme of the article, which is whether the simple receptor dimerization model explains all signal transduction following PRLR binding. PMID- 9867246 TI - Telomeres and telomerase in endocrine pathology. AB - Telomeres representing repetitive DNA sequences of chromosome ends are necessary for maintaining chromosomal integrity. The enzyme telomerase synthesizes de novo telomeric repeats and incorporates them onto the DNA 3'-ends of chromosomes. Stability of chromosome ends and activation of telomerase are elementary requirements for cell immortalization and tumor progression. The telomeric length and telomerase activity have been recently studied in several human neoplasms, including those of endocrine tissues. Assessment of telomerase activity may help to distinguish normal or hyperplastic from neoplastic tissues. Inhibition or inactivation of telomerase activity may provide novel strategies for cancer therapy. PMID- 9867247 TI - The thrifty genotype in type 2 diabetes: an unfinished symphony moving to its finale? AB - The basic premise of the thrifty gene hypothesis is that certain populations may have genes that determine increased fat storage, which in times of famine represent a survival advantage, but in a modern environment result in obesity and type 2 diabetes. The concept finds support in a unique animal model (Psammomys obesus) as well as among high type 2 diabetes susceptibility populations, such as North American Indians and South Pacific islanders. However, in some developing communities (e.g., Black South Africans) the thrifty phenotype hypothesis of perinatal malnutrition causing beta-cell dysfunction seems a better explanation, but this remains a contentious issue. Several genes have already been identified as candidates for the thrifty genotype, including those encoding proteins of the insulin-signaling and leptin pathways, as well as intermediary fat metabolism. Particular interest lies in the peroxisome-proliferator activated receptors. An innovative approach might be to focus on the "mirror image" of the thrifty genotype-congenital lipoatrophic diabetes mellitus, whose molecular defect remains enigmatic. We conclude that the genetic basis of the thrifty genotype probably derives from the multiplicative effects of polymorphisms at several sites mentioned above, rather than a single regulatory abnormality. PMID- 9867248 TI - Glucocorticoids modulate the biosynthesis and processing of prothyrotropin releasing-hormone (proTRH). AB - The thyrotropin- (TRH) releasing hormone precursor (26 kDa) undergoes proteolytic cleavage at either of two sites, generating N-terminal 15 kDa/9.5 kDa or C terminal 16.5/10 kDa intermediate forms that are processed further to yield five copies of TRH-Gly and seven non-TRH peptides. Glucocorticoids (Gcc) have been shown to enhance TRH gene expression in three different cell systems in vitro, an effect that occurs, at least in part, through transcriptional activation. Although this implies that an increase of TRH prohormone biosynthesis would take place, this had not been demonstrated as yet. We report here that the synthetic glucocorticoid dexamethasone (Dex) substantially elevated the de novo biosynthesis of the intact 26-kDa TRH prohormone and its intermediate products of processing in cultured anterior pituitary cells, an observation that is consistent with an overall upregulation of both the biosynthesis and degradation of the TRH precursor. We reasoned that Gcc may act not only at the transcriptional, but also at the translational/posttranslational level. To address this question we chose a different cell system, AtT20 cells transfected with a cDNA encoding preproTRH. Since TRH gene expression in these cells is driven by the CMV-IE promoter and not by an endogenous "physiological" promoter, these cells provide an ideal model to study selectively the effects of Gcc on the translation and posttranslational processing of proTRH without interference from a direct transcriptional activation of the TRH gene. Dex caused a significant 75.7% increase in newly synthesized 26-kDa TRH prohormone, suggesting that the glucocorticoid raised the translation rate. We then demonstrated that Dex treatment accelerated TRH precursor processing. Of interest, processing of the N- vs the C-terminal intermediate was influenced differentially by the glucocorticoid. Although the N-terminal intermediate product of processing accumulated, the C-terminal intermediate was degraded more rapidly. Consistent with these observations was the finding that the intracellular accumulation of the N-terminally derived peptide preproTRH 25-50 was enhanced, but levels of the C-terminally derived peptide preproTRH208-255 were reduced. Accumulation of TRH itself, whose five copies are N- and C-terminally derived, was also enhanced. We conclude that Gcc induce changes in the biosynthesis and processing of proTRH by increasing the translation rate and by differentially influencing the processing of N- vs C-terminal intermediates of the precursor molecule. These effects of Gcc at the translational and posttranslational levels result in an increase in TRH production accompanied by differential effects on the accumulation of N- and C terminal non-TRH peptides. PMID- 9867249 TI - Influence of cortisol on insulin- and insulin-like growth factor 1 (IGF-1) induced steroid production and on IGF-1 receptors in cultured bovine granulosa cells and thecal cells. AB - During stress, hyperactivity of the adrenal gland can directly and indirectly inhibit ovarian function. However, little evidence existed to support the notion that glucocorticoids could influence insulin-like growth factor 1 (IGF-1) action within the ovary. Therefore, the effect of cortisol on IGF-1-induced granulosa and thecal cell function was evaluated. Granulosa and thecal cells from bovine ovarian follicles were cultured for 2 d in the presence of 10% fetal calf serum and then cultured for an additional 2 d in serum-free medium with added hormones. Cortisol had little or no effect (p > 0.05) on IGF-1-induced progesterone production by granulosa cells from both small (1-5 mm) or large (> or =8 mm) follicles. Also, cortisol had little or no effect (p > 0.05) on basal, insulin-, or IGF-1-induced estradiol production by granulosa cells from small or large follicles, or on the number of IGF-1 receptors in granulosa cells from small follicles. Cortisol had no effect (p > 0.10) on insulin-induced granulosa cell numbers, but increased IGF-1-induced granulosa cell numbers. In thecal cells, doses of 1-100 ng/mL of cortisol increased (p< 0.05) insulin- and IGF-1-induced thecal cell numbers by 10-20%, progesterone production by 18-36%, and androstenedione production by two- to fourfold. The estimated dose of cortisol necessary to stimulate 50% of the maximum androstenedione production in the presence of IGF-1 was 7 ng/mL. In contrast, cortisol decreased (p < 0.05) the number of IGF-1 receptors in thecal cells by 45%. In conclusion, cortisol at physiological levels can directly influence ovarian follicular function in cattle, especially thecal androstenedione production. PMID- 9867250 TI - Prolactin-regulated apoptosis of Nb2 lymphoma cells: pim-1, bcl-2, and bax expression. AB - Lactogen-dependent Nb2 lymphoma cells, widely employed for studying prolactin (PRL) mitogenic mechanisms, are also useful for investigations of apoptosis in T lineage lymphocytes. Utilizing PRL-dependent Nb2-11 cultures, apoptosis regulatory genes were evaluated for participation in dexamethasone- (DEX) provoked cell death or its inhibition by PRL. Treatment of lactogen-starved, G1 arrested Nb2-11 cells with DEX (100 nM) activated apoptosis within 12 h evaluated by flow cytometric analysis of fragmented DNA. This effect was not associated with altered expression of bcl-2, bax, or pim-1. PRL (10 ng/mL), coincubated with DEX-treated cells, completely blocked DEX-induced apoptosis. This inhibition was associated with increased expression of bcl-2 and pim-1 mRNAs, genes reported to suppress apoptosis, within 2-6 h after addition of the hormone. Moreover, the increased transcription of bcl-2 and pim-1 was coupled to increases in their protein levels. The results suggest that bcl-2, bax, and pim-1 do not play a critical role in DEX-induced apoptosis in Nb2 cells. However, expression of bcl 2, together with pim-1, may have a role in mediating the antiapoptotic actions of PRL. PMID- 9867251 TI - Growth effects of regulatory peptides and intracellular signaling routes in human pancreatic cancer cell lines. AB - The intracellular events involved in normal pancreatic growth have been extensively investigated in response to cholecystokinin. Recent data indicate that tyrosine kinase, phospholipase D, phosphatidylinositol 3-kinase, and p42/p44 MAPK are stimulated in rat pancreatic acinar cells. Although we begin to understand the intracellular signaling pathways activated in normal pancreas, such information is not yet available in pancreatic cancer cells. This study was undertaken to identify the growth factors and hormones involved in cell proliferation of two human pancreatic cancer cell lines of ductal origin, the MIA PaCa-2, and PANC-1 cells, and to establish the intracellular events involved in the control of their growth. We demonstrated that FGF-2, IGF-1, cerulein, and gastrin but not FGF-1, HGF, secretin, and PACAP, stimulated proliferation of MIA PaCa-2 and PANC-1 cells. Autocrine factors such as gastrin and IGF-1 were also responsible for their proliferation. In response to EGF, FGF-2, IGF-1, cerulein, gastrin and bombesin, tyrosine kinase, and tyrosine phosphatase activities were stimulated in both cell lines. The close relationship established between cell growth and tyrosine kinase activation results from the observation that maximal growth stimulation paralleled with maximal enzyme activation and that genistein, the tyrosine kinase inhibitor, blocked cell growth and enzyme activation. The implication of PLD in growth-stimulated processes is doubtful since all growth factors and hormones tested failed to stimulate an already very active PLD activity. We finally observed a constitutive activity of p44 MAPK in both cell lines and of p42 in MIA PaCa-2 cells. However, p38 and p42 were stimulated in MIA PaCa-2 and PANC-1 cells, respectively, by all growth factors and hormones. PMID- 9867252 TI - Involvement of insulin-like growth factor-1 and its binding proteins in proliferation and differentiation of murine bone marrow-derived macrophage precursors. AB - Insulin-like growth factor 1 (IGF-1) and its binding proteins (IGFBPs) are involved in proliferation and differentiation of many cell types. In the present study, the involvement of IGF-1 and IGFBPs in proliferation and differentiation of murine bone marrow-derived macrophages (BMDM) was investigated. L929 conditioned media (LCM) containing abundant macrophage colony-stimulating factor CSF-1 were used to stimulate BMDM development from their bone marrow precursors. The alteration of IGF-1 and IGFBPs during LCM-induced BMDM proliferation and differentiation was first studied. The cells were cultured in RPMI complete media containing 20% LCM for different time periods and then incubated in serum-free media for 24 h. The supernatants were collected for Western ligand blotting and immunoblotting analyses, and the cell pellets for Northern blotting analyses. The mRNA level of IGF-1 increased in a time-dependent manner. An increase of IGFBP-4 accumulation in the conditioned media was also observed during this process. However the mRNA expression of IGFBP-4 remained constant, indicating a posttranscriptional regulation of IGFBP-4 secretion and/or stability. The effects of exogenous recombinant human IGF-1 (rhIGF-1) on BMDM proliferation and differentiation were further studied. Two IGF-1 analogs (long R3 IGF-1 and des [1 3] IGF-1) were also used in parallel with regular IGF-1 to indicate the involvement of IGFBPs in BMDM development. Cells were cultured in complete media containing 20% LCM for different time periods, and then incubated in serum-free media in the presence of rhIGF-1 or its analogs for 24 h. These three forms of IGF-1 all potentiated the proliferation of freshly isolated BMDM precursors (d 0). rhIGF-1 and long R3 IGF-1, but not des (1-3) IGF-1, continued to stimulate the cell proliferation on d 1. The effects of these three forms of IGF-1 on BMDM differentiation were investigated using mannose receptor expression as a marker. Long R3 IGF-1 and des (1-3) IGF-1, but not rhIGF-1, enhanced BMDM differentiation on d 4. The different effects of rhIGF-1 and its analogs on BMDM differentiation suggest that the accumulation of IGFBP-4 in BMDM development might have an inhibitory effect on IGF-1 actions by sequestering free IGF-1. PMID- 9867253 TI - c-Fos dimerization with c-Jun represses c-Jun enhancement of androgen receptor transactivation. AB - The transcriptional activity of the human androgen receptor (hAR), like other nuclear receptors, is dependent on accessory factors. One such factor is c-Jun, which has been shown to have a selective function of mediating androgen receptor dependent transactivation. This c-Jun activity is inhibited by c-Fos, another protooncoprotein that can dimerize with c-Jun to form the transcription factor AP 1. Here we show that c-jun mediates hAR-induced transactivation from the promoter of the androgen-regulated gene, human kallikrein-2 (hKLK2), and c-Fos blocks this activity. Using c-Fos truncation mutants and measuring hKLK2-dependent transcription, we have determined that the bZIP region of c-Fos is required and sufficient for inhibiting c-Jun enhancement of hAR transactivation. Further truncation analysis of the bZIP shows that the c-Fos dimerization function, mediated through the leucine zipper, is essential for the negative activity, whereas DNA binding, mediated through the basic region, is dispensable. These results suggest that heterodimerization by c-Fos with c-Jun blocks c-Jun's ability to enhance hAR-induced transactivation. PMID- 9867254 TI - Synaptophysin immunoreactivity in the rat pituitary: alterations after 6 hydroxydopamine treatment. AB - Synaptophysin (SN) is a synaptic-vesicle-associated membrane protein whose presence is indicative of intact, functional synapses. This study examines the presence of SN in pituitary gland innervation after neurotoxin-induced denervation followed by reinnervation. Immunostaining of rat pituitary neurointermediate lobe tissue for SN reveals a pattern of dot-like densities in the intermediate lobe and intensely stained dispersed regions in the neural lobe of normal animals. In rats treated with 6-hydroxydopamine (6-OHDA), a catecholamine neurotoxin, by peripheral injection, there is a significant depletion of the SN immunostaining in the intermediate lobe, as well as a significant reduction of SN immunoreactivity in the neural lobe, in animals studied 1 wk after drug treatment, with computer analysis of the tissue sections. At 3 wk after 6-OHDA, there is a partial recovery of immunoreactivity for SN in the neural lobe in many tissue sections, and the intermediate lobe also contains only relatively sparse staining for the synaptic protein. Computer analysis revealed that at 3 wk after 6-OHDA, both lobes still had reduced SN immunoreactivity, but the difference in levels measured did not achieve statistical significance. These results contrast with the prior finding of significant recovery of immunoreactivity for GAP-43, a growth and regeneration associated protein, in intermediate lobe innervation of rats treated with the same drug regimen. We suggest that 6-OHDA treatment damages synaptic vesicle integrity in both the intermediate and neural lobes of the pituitary, and that recovery is in progress, but not complete at 3 wk after the drug is administered. PMID- 9867255 TI - Effect of environmental estrogens on IL-1beta promoter activity in a macrophage cell line. AB - Environmental estrogens or estrogen disrupters have recently received a great deal of attention because of their potential health impact on reproductive tissues. Few, if any, studies have been made on the impact of these compounds on the immune system. We sought to determine the activities of various environmental estrogens on the modulation of the interleukin-1beta (IL-1beta) gene in a model monocytic cell line, hER + IL-1beta-CAT+. This cell line stably transfected with the human estrogen receptor, and an IL-1beta promoter construct fused to the CAT reporter gene allows us to monitor the effect of estrogenic compounds on IL-1beta promoter activity. 17beta-estradiol (E2) markedly enhanced lipopolysaccharide- (LPS) induced IL-1beta promoter-driven CAT activity in a dose-dependent manner. The mycotoxins alpha-zearalenol and zearalenone both exhibited full agonist activity, but at lower potencies, with EC50 values of 1.8 and 54 nM, respectively, compared with E2 at 0.5 nM. In addition, genistein was a very low potency agonist, having an EC50 of 1.5 microM. Similar to the E2 response, the slope factors for alpha-zearalenol, zearalenone, and genistein were close to 3.0, suggesting positive cooperativity in the estrogenic response. The activity of the mycotoxins appeared to be mediated through the estrogen receptor, since both the antiestrogens H1285 and ICI 182,780 effectively inhibited their agonist activity in a dose-dependent manner. Representative environmental estrogenic compounds both from plant and industrial sources were also tested. Unlike the mycoestrogens, none of the compounds, with the exception of genistein, synergized with LPS to enhance IL-1beta promoter activity. When tested for antiestrogenic activity, the industrial compound 4-octylphenol was able to antagonize the response to E2; however, the response was three orders of magnitude less potent than H 1285. Naringenin, a plant flavonoid, showed little or no ability to antagonize the response to E2. Overall, the results show that some environmental estrogens that display agonist activity in reproductive tissue also have an effect on IL-1 gene expression in hemopoietic-derived tissue. PMID- 9867256 TI - Chronic ethanol drinking and food deprivation affect rat hypothalamic-pituitary thyroid axis and TRH in septum. AB - Because chronic ethanol ingestion may perturb thyroid function, we evaluated the effect of 4-wk of oral 10% ethanol ingestion on the hypothalamic-pituitary thyroid (HPT) axis and septal thyrotropin-releasing hormone (TRH) in 200-g male Wistar rats. Animals were divided into three groups: absolute control receiving tap water and food ad libitum; ethanol group receiving food ad libitum and 10% ethanol as the sole source of drinking fluid; pair-fed group receiving tap water and an amount of food corresponding to the consumption of ethanol group. After 4 wk of treatment, the body weight of the ethanol group was 7% and of the pair-fed rats 19% lower than that of the absolute controls. Both chronic ethanol treatment and food deprivation produced a decrease in plasma thyroid-stimulating hormone (TSH). Pair-fed rats also had a lower plasma T3. Type I iodothyronine 5' deiodinase activity in the liver was increased in the pair-fed and even more in the ethanol-treated group. The content and secretion in vitro of TRH from the hypothalamic paraventricular nucleus and median eminence were unchanged. TRH content in the septum was increased in both the ethanol and pair-fed groups. TRH secretion from the septum in vitro was lower in the pair-fed, but unchanged in the ethanol group. These data suggest that 4-wk of peroral ethanol intake affects thyroid function mostly at the extrahypothalamic level and that there is a contribution of concomitant food deprivation. Both ethanol treatment and food deprivation increased TRH content in the septum. PMID- 9867257 TI - Comparison of the Block and the Willett self-administered semiquantitative food frequency questionnaires with an interviewer-administered dietary history. AB - The performances of two commonly used diet instruments, the Block and the Willett food frequency questionnaires, were compared with a longer, interviewer administered diet history. Participants in a case-control study on diet and colon cancer were interviewed between 1990 and 1994 in northern California, Utah, and Minnesota by trained nutritionists using a validated diet history. Two separate subsamples of participants were asked to complete either the Block or the Willett questionnaire exactly 5 days after they completed the original diet history. Data were analyzed separately by subsample comparing either the Block or the Willett questionnaire with the original diet history by using means, correlations, quintile agreement, and odds ratios for the relation between several nutrients and colon cancer. The Block and the Willett questionnaires generally provided lower absolute intake estimates than did the original diet history; however, the Block questionnaire underestimated more than did that by Willett. Both correlations and quintile agreement were slightly better for the Willett questionnaire than for that by Block when compared with the original diet history. In general, point estimates obtained from either the Block or the Willett questionnaire fell within the confidence intervals of the estimates of the odds ratios obtained from the original diet history, and no real difference in significance levels appeared. Although the Block and Willett questionnaires differed slightly from each other and from our original diet history in estimating absolute nutrients and ranking or classifying individuals, they were very similar in their ability to predict disease outcome. PMID- 9867258 TI - Comparison of food frequency questionnaires: the reduced Block and Willett questionnaires differ in ranking on nutrient intakes. AB - Food frequency questionnaires, major tools in epidemiologic studies, are often criticized for biased and imprecise intake estimates. The aim of this study was to compare the performance of two widely used food frequency questionnaires, a reduced 60-item Block questionnaire and a 153-item Willett food frequency questionnaire, relative to three 24-hour recalls administered by telephone. The dietary data were collected in 1991 from a group of healthy women age 25-49 years (n=101) during the baseline period of a weight-loss intervention study in Minneapolis, Minnesota. Total energy and macro- and micronutrient intakes were compared across methods by using four analytic approaches: comparison of means and correlation coefficients, regression analysis, and estimation of percent agreement between each questionnaire and recalls. The Block instrument showed an overall underestimation bias, but was more successful in categorizing individuals on percent energy from fat and carbohydrate intakes than was the Willett instrument. The Willett instrument showed no overall underestimation bias and was more successful in classifying individuals on vitamin A and calcium intakes. Diverging performance characteristics of diet assessment methods have an implication for the design of studies, interpretation of results, and comparison of findings across studies. PMID- 9867259 TI - Invited commentary: comparison of food frequency questionnaires. PMID- 9867260 TI - Invited commentary: comparison of the Block and the Willett food frequency questionnaires. PMID- 9867261 TI - Snippets from the past: 70 years ago in the Journal. PMID- 9867262 TI - Clusters of marijuana use in the United States. AB - In this study, the authors test for and estimate the clustering of marijuana use within United States neighborhoods, making use of data from annual nationally representative household sample surveys conducted during the period 1990-1995. A recently developed statistical method, alternating logistic regression, was used to quantify the clustering of marijuana users in neighborhoods. The resulting estimates of pairwise odds ratios ranged from 1.3 (95% confidence interval 1.22 1.42) for the lifetime history of marijuana use to 2.0 (95% confidence interval 1.6-2.6) for recent sharing of marijuana from one person to another. Exploratory analysis showed a slight decrease of clustering effects after adjustment for individual-level covariates: age, sex, race, education, annual family income, and history of tobacco use. Nevertheless, the main factors that account for clustering remain to be determined. Alternating logistic regression provided useful estimates of marijuana use clustering and can be used to estimate clustering of the other drug-related behavior, including sharing of needle injection equipment and other human immunodeficiency virus risk behaviors. As a form of multilevel analysis, the alternating logistic regression can accommodate shared, community-level characteristics that might influence drug taking (e.g., collective efficacy), as well as individual-level covariates, such as age and sex. PMID- 9867263 TI - Neutropenia is a risk factor for gram-negative bacillus bacteremia in human immunodeficiency virus-infected patients: results of a nested case-control study. AB - A previous cohort study demonstrated a relation between neutropenia and bacteremia due to gram-negative bacilli among patients infected with human immunodeficiency virus (HIV). To explore further the relation between neutropenia and bacteremia due to Escherichia coli, Klebsiella pneumoniae, or Pseudomonas aeruginosa among HIV-infected patients, controlling for confounding factors, the authors conducted a nested case-control study with matching and risk-set sampling of controls. The cohort included 1,645 HIV-infected patients followed at the University of California, San Diego, Medical Center in San Diego, California, between 1991 and 1995. Absolute neutrophil count (ANC) was summarized as mean ANC during the 7-day interval preceding the index date of bacteremia. Covariates were ascertained by medical record review. The matching ratio was 6:1 (controls:cases). Odds ratios were estimated using conditional logistic regression. Forty-four incident cases of bacteremia were identified. After adjustment for covariates, the estimated odds ratio for the effect of neutropenia (ANC=500 vs. >500/microl) during the 7 days preceding the index date was 8.1 (95% confidence interval confidence interval 1.5-43.1). The rate of bacteremia due to E. coli, K. pneumoniae, or P. aeruginosa is increased eightfold if the average current-week ANC is less than or equal to 500/microl compared with more than 500/microl. PMID- 9867264 TI - Validation of cause-of-death certification for outpatient cancers: the contrasting cases of melanoma and mycosis fungoides. AB - The reported incidence of cancers typically diagnosed in the outpatient setting may underestimate their true frequency, but the validity of mortality estimates from many of these causes has not been studied in detail. The authors sought to evaluate the validity of mortality estimation from death certificates for two such cancers, melanoma and mycosis fungoides (a cutaneous lymphoma), using routinely collected cancer registry data for 1973-1994 from the Surveillance, Epidemiology, and End Results program. Their method estimates that 93% of the deaths attributable to melanoma were certified as due to melanoma but that only 60% of the deaths attributable to mycosis fungoides were so certified. Evaluation of the accuracy of cause-of-death certification in this manner is helpful in the interpretation of mortality statistics. PMID- 9867265 TI - Body mass index, waist/hip ratio, and coronary heart disease incidence in African Americans and whites. Atherosclerosis Risk in Communities Study Investigators. AB - To study the relation of the amount and distribution of body fat with incident coronary heart disease in two ethnic groups, the authors analyzed prospective data from the Atherosclerosis Risk in Communities Study. Among 14,040 participants aged 45-64 years and free of coronary disease at baseline in 1987 1989, we identified 398 events through 1994, an average of 6.2 years of follow up. Among African-American women, the multivariable-adjusted relative risks of coronary heart disease across quartiles of body mass index were 1.0, 1.91. 1.54, and 2.15 (p for trend=0.27), and those for waist/hip ratio were 1.0, 2.07, 2.33, and 4.22 (p for trend=0.02). Among African-American men, these respective relative risks were 1.0, 1.03, 0.83, and 1.20 (p for trend=0.76) for body mass index and 1.0, 1.08, 1.87, and 1.68 (p for trend=0.06) for waist/hip ratio. Relative risks for whites were generally similar to those for African Americans. Relative risks were stronger for never smokers than for the overall cohort. Unlike some previous studies, our results suggest that Africa Americans, like whites, are not spared from the coronary heart disease risks accompanying obesity. PMID- 9867267 TI - Impact of prenatal diagnosis and elective termination on the prevalence of selected birth defects in Hawaii. AB - This study examined the effect of prenatal diagnosis and elective termination on the prevalence of neural tube defects, oral clefts, abdominal wall defects, and chromosomal anomalies in Hawaii by using actively ascertained surveillance data collected between 1987 and 1996 by the Hawaii Birth Defects Program. Because the Program has nearly universal access to prenatal diagnostic information and to follow-up data on elective terminations, Hawaii is an ideal setting in which to study their effects on prevalence rates of birth defects. Except for oral clefts, a large proportion of the defects studied were prenatally diagnosed: anencephaly (87%), spina bifida (62%), encephalocele (83%), cleft palate (0%), cleft lip with or without cleft palate (14%), omphalocele (60%), gastroschisis (76%), Down syndrome (43%), trisomy 18 (61 %), and trisomy 13 (40%). The effect of elective terminations on the birth prevalence rates for most of these birth defects was significant. Including electively terminated cases in the calculations of birth prevalence rates increased the rates by more than 50% for five of the 10 birth defects studied. PMID- 9867268 TI - Geocoding and linking data from population-based surveillance and the US Census to evaluate the impact of median household income on the epidemiology of invasive Streptococcus pneumoniae infections. AB - The emergence of drug-resistant Streptococcus pneumoniae poses new clinical challenges and may also reflect a change in the epidemiology of S. pneumoniae infections. A variety of studies have shown that drug-resistant S. pneumoniae infections are linked to antimicrobial use. It has been hypothesized that persons of high socioeconomic status are at increased risk for a drug-resistant infection because of greater access to antimicrobial drugs. To assess whether median household income is associated with increased risk of penicillin-nonsusceptible S. pneumoniae infections, the authors geocoded and linked data from population based surveillance for invasive pneumococcal disease with data from the 1990 US Census. Among invasive pneumococcal isolates from Atlanta, Georgia, in 1994, increasing proportions of penicillin-nonsusceptible isolates were associated with higher median household incomes (chi2 for trend, 15.17; p=0.002). Despite higher rates of invasive pneumococcal disease among blacks and persons who resided within lower median household income areas, white patients in areas with higher median household income had a higher risk of being infected with strains that were not susceptible to penicillin (Wilcoxon rank sum, Z=2.66, p=0.008). These findings demonstrated the utility of geocoding and US Census data in describing the epidemiology of drug-resistant S. pneumoniae and also provided more evidence that socioeconomic factors may influence the development of drug resistance. PMID- 9867266 TI - International variability in ages at menarche, first livebirth, and menopause. World Health Organization Collaborative Study of Neoplasia and Steroid Contraceptives. AB - The occurrences and timing of reproduction-related events, such as menarche, first birth, and menopause, play major roles in a woman's life. There is a lack of comparative information on the overall patterns of the ages at and the timing between these events among different populations of the world. This study describes the variability in reproductive factors across populations in Europe, the Americas, Asia, Australia, and Africa. The study sample consisted of 18,997 women from 13 centers in 11 countries interviewed between 1979 and 1988 who comprised the control group in a World Health Organization international, multicenter case-control study of female cancers. All were surveyed with the same questionnaire and methodology. Overall, a typical woman in this study reached menarche at age 14 years and delivered her first live child 8 years later, at age 22. She was 50 years old at natural menopause and had had 36 years of reproductive life. The median ages at menarche varied across centers from 13 to 16 years. For all centers, the median age at first livebirth was 20 or more years, with the largest observed median (25 years) occurring in China. The median delay from menarche to first livebirth ranged from 5 to 11 years. Among the centers, the median age at natural menopause ranged between 49 and 52 years. In most populations, younger women had a first birth at a later age than did older women. This tendency was more accentuated in some populations. These results reveal, perhaps for the first time, the variability of reproductive histories across different populations in a large variety of geographic and cultural settings. Except for menopause, international variability is substantial for both biologically related variables (age at menarche) and culturally related variables (age at first birth). There is a generational effect, characterized by more variability of age at first birth and delay to first birth in the younger than in the older generations. PMID- 9867269 TI - Hydatid disease: biology, pathology, imaging and classification. AB - Rupture and the sequellae of rupture are more important than the mass effect of hydatid cysts, except in the brain where the mass effect by itself has severe consequences. The biology of hydatid disease, including the complex interaction between primary and secondary hosts, is reviewed. The hydatid cyst always starts as a fluid-filled, cyst-like structure (Type I) which may proceed to a Type II lesion if daughter cysts and/or matrix develop. In some instances the Type II lesion becomes hypermature and due to starvation dies to become a mummified, inert calcified Type III lesion. Type I and II lesions may undergo three types of rupture: contained, communicating and direct. Contained rupture is clinically silent, but communicating rupture may cause biliary obstruction and evacuation or infection of the cyst. Direct rupture has the greatest clinical consequences which include anaphylaxis, dissemination of hydatid disease (secondary hydatosis) within the host, and bacterial infection of the pericyst cavity. The clinical implications of the hydatid disease at different stages are discussed. A plea is made for the development of an international medical hydatid registry employing uniform nomenclature and consistent reporting in order to allow more rational comparisons of different types of management. PMID- 9867270 TI - Pictorial review: Subperitoneal spaces of the broad ligament and sigmoid mesocolon--imaging findings. AB - In this report, we illustrate the imaging findings of diseases in the subperitoneal space of the broad ligament and sigmoid mesocolon and discuss the pathways of subperitoneal spread of disease. The subperitoneal space of the broad ligament and sigmoid mesocolon is continuous with that of the parietal peritoneum overlying the pelvic wall. Extraperitoneal diseases originating in the pelvis can extend, via the abdominopelvic vasculature or fascial-defined compartment, superiorly into the retroperitoneal compartments of the abdomen. PMID- 9867271 TI - Pictorial review: radiology of the sphenoid bone. AB - The sphenoid bone is located in the central skull base and forms part of the floor of the anterior and middle cranial fossae. The optic foramen, superior orbital fissure, foramen rotundum, foramen ovale and foramen spinosum are found within this complex bone. These foramina form important transition zones between intracranial and extracranial structures. Imaging plays a central role in delineating lesions within the sphenoid bone and the associated fissures and neural foramina. The purpose of this pictorial review is to highlight the pertinent normal and pathological anatomy of the sphenoid bone. PMID- 9867272 TI - Is minimal preparation computed tomography comparable with barium enema in elderly patients with colonic symptoms? AB - The barium enema (BE) examination in the elderly is often an unsatisfactory investigation. Recent papers have proposed computed tomography (CT) as an alternative method of investigation. They have used a variety of preparation techniques prior to the CT study such as bowel preparation, rectal air insufflation, tap water enemas and intravenous contrast medium. We wished to devise a minimal preparation, minimal supervision technique that could realistically replace the standard BE. One hundred and eighteen elderly patients admitted with symptoms referable to the large bowel had an unenhanced CT following oral contrast medium commenced the previous evening, followed at an interval by a BE performed to a standard technique. Both techniques congruently gave negative reports in 66 (68.8%) individuals. Ten tumours were confidently diagnosed on CT. One annular tumour of the sigmoid colon and one case of colitis were diagnosed on BE only and the severity of a diverticular stricture was underestimated on CT. CT raised the possibility of lesions in 3 (3.1%) patients where the BE was negative. Ten (8.5%) patients had significant abnormalities identified outside the colon using CT. Seven (5.9%) barium enemas were abandoned or severely limited. Minimal preparation, minimal supervision CT is a practical alternative to BE in the frail elderly patient. Radiologists and referring clinicians should be aware that occasionally lesions will not be identified on CT but CT has the advantage that other significant pathology unrelated to the colon may be detected. PMID- 9867273 TI - Can pulmonary venous hypertension be graded by the chest radiograph? AB - AIM: The chest radiograph is widely used for the diagnosis and follow-up of patients with pulmonary venous hypertension (PVH) caused by rheumatic heart disease. Observer accuracy and interobserver agreement on the radiographic diagnosis of PVH is rarely assessed. We have studied this accuracy and agreement in 120 consecutive patients. METHODS AND RESULTS: Patients were selected on the basis of mean pulmonary capillary wedge pressures (PCWP) obtained in the basal state. A chest radiograph was obtained within the 24 h before catheterization and was assessed by two observers blinded to the diagnosis and PCWP. PVH was graded as absent (PCWP < 12 mmHg); mild (PCWP 12-19 mmHg; redistribution of blood flow and hilar haze); moderate (PCWP 20-25 mmHg; septal oedema, pleural effusion) and severe (PCWP > 25 mmHg; alveolar oedema). Observer and interobserver agreement was quantified by using the Kappa (kappa) statistic. The radiological assessment coincided with the measured PCWP in 52% of the patients according to observer 1 and 43% of the patients according to observer 2 (kappa 0.36 and 0.24), indicating poor agreement with haemodynamic values. Interobserver agreement was poor (kappa 0.3-0.35). Observer accuracy and interobserver agreement increased when the radiographic criteria were used to distinguish patients with normal PCWP from those with abnormal pressures when 85% and 78% of radiographs were correctly classified (kappa 0.6 and 0.5) by observers 1 and 2, respectively. Overall agreement was significant (kappa 0.6). In differentiating patients with a PCWP > 20 mmHg from those with a PCWP < 20 mmHg, 78% and 75% of radiographs were correctly classified, giving kappa values of 0.6 and 0.5. Overall observer agreement was significant (kappa 0.6). CONCLUSION: The chest radiograph is useful in detecting the presence of PVH and in distinguishing significant elevations in PCWP (> 20 mmHg) from mildly elevated PCWP. There is poor correlation between haemodynamic and radiographic findings when more precise grading of the severity of PVH is attempted. PMID- 9867274 TI - Multiple symmetric lipomatosis in the Chinese: ultrasound, CT and MR imaging. AB - We report four cases of multiple symmetric lipomatosis in the Chinese population. We believe that multiple symmetric lipomatosis in the Chinese is not uncommon and may be related to the increasing incidence of alcoholism. The ultrasound appearances of these lipomatous masses are presented for the first time. Heterogeneous echogenic masses with fine fibrous strands that insinuate around fascial planes, lymph nodes and vascular tissues are typical findings. Unlike diseases affecting the Western population, multiple symmetric lipomatosis in the Chinese appears to be limited to the head and neck. PMID- 9867275 TI - Fibrosing mediastinitis: CT and MR findings. AB - The aim of this study is to present the computed tomography (CT) and magnetic resonance (MR) findings in three cases of fibrosing mediastinitis, probably secondary to tuberculosis. Two patients had diffuse involvement of the mediastinum and one patient presented with a posterior mediastinal mass. On CT, the patients with diffuse involvement showed infiltrating soft-tissue masses with multiple foci of dense calcifications. One of these lesions enhanced mildly with contrast and one did not enhance. On MR examination, diffuse masses were of heterogeneous, intermediate and low signal on T1-weighted images (n = 2) and were of homogeneous low signal (n = 1) or heterogeneous (n = 1) signal intensity on T2 weighted images. After administration of Gd-DTPA, these mediastinal masses showed heterogeneous enhancement. The posterior mediastinal mass showed homogeneous pronounced enhancement on CT and was homogeneous with intermediate T1 signal intensity and hypointense T2 signal intensity. The CT findings of an infiltrative or well-defined mediastinal mass with scattered calcifications should suggest the diagnosis of fibrosing mediastinitis in the appropriate clinical setting. Although the appearance of calcification on MR are not specific, this technique provides further information than CT especially in those cases in which the lesions appear noncalcified, because MR imaging revealed areas of low signal intensity characteristic of fibrous tissue. PMID- 9867276 TI - RCR 1997 scientific meeting: audit poster prize winners. PMID- 9867277 TI - Case report: Dissecting aneurysm of the hepatic artery. PMID- 9867278 TI - Case report: Biliary stent placement via percutaneous non surgical cholecystostomy. PMID- 9867279 TI - Case report: Serous cystadenocarcinoma of the retroperitoneum: CT and sonographic appearance. PMID- 9867280 TI - Case report: Intradiaphragmatic bronchogenic cyst. PMID- 9867281 TI - Case report: Primary osteosarcoma of the breast: imaging and histological features. PMID- 9867282 TI - Technical report: Initial experiences with an experimental solid-state universal digital X-ray image detector. AB - This paper presents a brief technical evaluation and first review of clinical experiences with an experimental direct digital X-ray image detector designed to support both dynamic and snap-shot imaging. Derivatives of this type of image detector can potentially fulfil the majority of the fluoroscopic and radiographic imaging requirements of clinical radiology departments, and initial results suggest that imaging systems using the new technology will provide a high quality dose-efficient solution to the search for a universal digital X-ray image detector. PMID- 9867283 TI - Technical report: A low cost technique for vessel sizing in angioplasty. AB - Vessel measurement in angiography is important for balloon and stent sizing. The most accurate techniques involve calibrating the angiographic image with a catheter with radioopaque markers, but these are expensive. A new method of vessel sizing for angioplasty and stenting is described: the guidewire is moved within the vessel through a set distance, and this movement is shown by digital subtraction. Testing with phantoms shows this to be simple, accurate and reproducible. PMID- 9867284 TI - CT demonstration of calcified metastatic mediastinal lymph nodes. PMID- 9867285 TI - Radiation doses to patients undergoing pelvimetry. PMID- 9867286 TI - Percutaneous sialography. PMID- 9867287 TI - Radiographic detection of achalasia. PMID- 9867288 TI - Metformin and contrast media. PMID- 9867289 TI - Gallstone ileus: CT findings. PMID- 9867290 TI - On the behavioral characteristics of loud-music listening. AB - To provide insight into the behavioral characteristics of people who listen excessively to loud music, the 32-item Northeastern Excessive Music Listening Survey was developed and administered to 90 subjects. Results indicate that 8 of the 90 subjects scored within a range that would suggest the presence of a maladaptive pattern of music-listening behavior similar to that exhibited by substance abusers. Implications for further research and models of treatment are discussed. PMID- 9867291 TI - The effect of masker interaural time delay on the masking level difference in children with history of normal hearing or history of otitis media with effusion. AB - OBJECTIVE: To determine the relation between the masking level difference (MLD) and the interaural time delay of the stimulus in children with a history of normal hearing and with a history of otitis media with effusion (OME). DESIGN: MLDs for a 500 Hz pure tone presented in a 100 Hz-wide masking noise were determined as a function of the interaural delay of the masker (six interaural delays between -726 microsec and +998 microsec were examined). For the masker with zero interaural delay, the signal was presented either interaurally in-phase or 180 degrees out of phase. For the masker delay conditions, the signal was given the same interaural delay as the masker and then was inverted interaurally. All children had normal audiograms at the time of testing. Ten children had a history of normal hearing and seven children had a history of OME. RESULTS: Similar to what has been found previously in adults, children with a history of normal hearing showed the maximum MLD (approximately 16 dB) for an interaural time delay of 0 microsec, with the MLD decreasing as a function of interaural time delay (by as much as 4 to 5 dB for the extreme delays). Children with a history of OME had significantly smaller MLDs than normal for the three smallest interaural delays but did not differ significantly from normal at the three largest interaural delays. CONCLUSIONS: The form of the function relating masker interaural time delay to MLD magnitude is adult-like by age 6 yr. The function indicates a binaural advantage for the processing of sound near auditory midline. This advantage is less apparent in children having a history of OME. PMID- 9867292 TI - Transient evoked otoacoustic emissions in patients with normal hearing and in patients with hearing loss. AB - OBJECTIVES: 1) To evaluate transient evoked otoacoustic emission (TEOAE) test performance when measurements are made under routine clinical conditions. 2) To evaluate TEOAE test performance as a function of frequency and as a function of the magnitude of hearing loss. 3) To compare test performance using univariate and multivariate approaches to data analyses. 4) To provide a means of interpreting clinical TEOAE measurements. DESIGN: TEOAEs were measured in 452 ears of 246 patients. All measurements were made after acoustic immittance assessments, which were used to demonstrate that middle-ear function was normal at the time of the TEOAE test. TEOAE amplitudes and signal to noise ratios (SNRs), analyzed into octave bands centered at 1, 2, and 4 kHz, were compared with the pure-tone threshold at the same frequencies. Data were analyzed with clinical decision theory, cumulative distributions, discriminant analyses, and logistic regressions. RESULTS: Using univariate analysis techniques, TEOAEs accurately identified auditory status at 2 and 4 kHz but were less accurate at 1 kHz. Test performance was best when audiometric thresholds between 20 and 30 dB HL were used as the criteria for normal hearing. TEOAE SNR resulted in better test performance than did TEOAE amplitude alone; this effect decreased as frequency increased. Multivariate analysis methods resulted in better separation between normal and impaired ears than did univariate approaches, which relied on only TEOAE amplitude or SNR when test frequency band and audiometric frequency were the same. This improvement in test performance was greatest at 1 kHz, decreased as frequency increased, and was negligible at 4 kHz. CONCLUSIONS: TEOAEs can be used to identify hearing loss in children under routine clinical conditions. Univariate tests accurately identified auditory status at mid and high frequencies but performed more poorly at lower frequencies. The decrease in performance as frequency decreases may be a result of increased noise at lower frequencies but also may be due to properties of the measurement paradigm ("QuickScreen," high-pass filter at 0.8 kHz), which would not be ideal for recording energy around 1 kHz. The improvement in test performance when SNR was used and the interaction of this effect with frequency, however, would be consistent with the view that test performance in lower frequencies is at least partially influenced by the level of background noise. Multivariate analysis techniques improved test performance compared with the more traditional univariate approaches to data analysis. An approach is provided that allows one to assign measured TEOAE amplitudes, SNRs, or outputs from multivariate analyses to one of three categories: response properties consistent with normal hearing; results consistent with hearing loss; hearing status undetermined. PMID- 9867293 TI - Effects of background noise on click-evoked otoacoustic emissions. AB - OBJECTIVE: To investigate the effect of increased levels of background noise on click-evoked otoacoustic emission (CEOAE) recordings and to compare the effectiveness of the default CEOAE program with the QuickScreen CEOAE program in increased levels of noise, using an Otodynamics ILO88 recording device. DESIGN: The right ears of 40 young adult women with normal hearing were assessed using CEOAEs under four different noise conditions and with two different methods of data collection. The noise conditions were in quiet, 50 dB A, 55 dB A, and 60 dB A of white noise. Data were collected at each noise level in the default mode and also using the ILO88 QuickScreen program. RESULTS: There was a significant change in a number of important CEOAE output parameters with increased noise. In the default mode, mean whole wave reproducibility was 89.2% in quiet but declined to 85% with 50 dB A of white noise, 65% at 55 dB A and 20% at 60 dB A. The QuickScreen program proved more robust to the effects of noise than the default. In that mode, mean whole wave reproducibility was 91.7% in quiet, 92.5% with 50 dB A of white noise, 82.5% at 55 dB A and 45% at 60 dB A. CONCLUSIONS: The findings of the study indicate ambient noise levels for accurate CEOAE recording should not exceed 50 to 55 dB A of noise and alternatives to the default program should be considered in non-sound-treated situations. PMID- 9867294 TI - Distortion product otoacoustic emission and auditory brain stem response measures of pediatric sensorineural hearing loss with islands of normal sensitivity. AB - OBJECTIVE: To assess the accuracy of: 1) distortion product otoacoustic emission (DPOAE) measures for the identification of frequencies at which auditory sensitivity is normal or near normal; and 2) click and nonmasked tone burst evoked auditory brain stem response (ABR) thresholds for behavioral threshold estimation for children with sensorineural hearing loss characterized by islands of normal sensitivity. DESIGN: DPOAEs and ABRs were recorded from five hearing impaired and eight normal-hearing pediatric ears. The accuracy with which DPOAEs permitted identification of frequencies at which elevated hearing thresholds were present was examined. ABR and pure-tone threshold differences for the impaired ears were calculated. RESULTS: For three of the five hearing-impaired ears, significant impairments would have been missed based on click-evoked ABR thresholds. One of those hearing-impaired ears provided an essentially normal 500 Hz tone burst-evoked ABR threshold as well. Four of the hearing-impaired ears provided a 500 Hz tone burst-evoked ABR threshold within 10 dB of the respective pure-tone threshold. However, click-evoked ABR and 500 Hz tone burst-evoked ABR threshold data did not adequately delineate the hearing loss configuration for hearing aid frequency response selection. DPOAEs were present at three out of four frequencies from 1000 to 4000 Hz at which sensitivity was normal or near normal (< or =25 dB HL) and absent at 10 out of 11 frequencies at which sensitivity was impaired. The use of DPOAEs to identify frequencies at which sensitivity was normal and the use of tone burst ABR thresholds at frequencies where DPOAEs were absent provided a better estimate of these pure-tone audiograms than was provided by click-evoked and 500 Hz tone burst-evoked ABR thresholds. PMID- 9867295 TI - Investigation of the profile of hearing aid performance in experienced hearing aid users. AB - OBJECTIVE: A shortened version of the 66-item Profile of Hearing Aid Performance (PHAP), consisting of the 24 items from the Abbreviated Profile of Hearing Aid Benefit, was evaluated as an alternative to the full PHAP questionnaire as a measure of hearing aid performance. Objectives were to: 1) statistically determine factors underlying Abbreviated PHAP (APHAP) scores; 2) recommend modifications to the questionnaire, if indicated by the factor analysis; 3) compare scores for the PHAP and APHAP; and 4) determine the suitability of the abbreviated questionnaire for New Zealand hearing aid users. The relationship between hearing aid performance and subjective variables and other measures of hearing aid success also was investigated. DESIGN: PHAP data, hearing aid satisfaction ratings, and reported daily hearing aid use were obtained from experienced adult hearing aid users. Factor analysis was carried out for the 24 APHAP items, and two items were excluded because of low factor loadings. After this modification, APHAP and PHAP scores were compared. Pearson's correlation values were determined for PHAP and APHAP data and degree of hearing loss, hearing aid satisfaction, and hours of hearing aid use. RESULTS: APHAP hearing aid performance was better described as three factors rather than four subscales. PHAP and APHAP results were consistent with previous studies and showed that hearing aid performance was best for easy listening situations and poorest for noisy and/or reverberant conditions. Similar trends were seen for PHAP and APHAP data. APHAP scores were correlated with hours of hearing aid use and overall hearing aid satisfaction. CONCLUSIONS: A shortened, 22-item APHAP is a preferred alternative to the full PHAP, producing data representing several dimensions of hearing aid performance. Modified APHAP scores can be used together with overall satisfaction and estimates of daily hearing aid use to measure success with hearing aids. PMID- 9867296 TI - The identification of speech in noise by cochlear implant patients and normal hearing listeners using 6-channel signal processors. AB - OBJECTIVE: To compare the recognition of vowels and sentences in noise by cochlear implant patients using a 6-channel, continuous interleaved sampling (CIS) processor and by normal-hearing subjects listening to speech processed in the manner of the implant processor and output as six amplitude-modulated sine waves. DESIGN: Subjects, 11 normal-hearing listeners and 7 cochlear implant patients, were presented natural vowels produced by men, women, and girls in /hVd/ context and sentences from the Hearing In Noise Test (HINT) lists at +15, +10, and +5 dB signal to noise ratio (SNR) for identification. Stimuli for the normal-hearing subjects were preprocessed through a simulation of a 6-channel implant processor and were output as the sum of sinusoids at the center frequencies of the analysis filters. RESULTS: For the multitalker vowels, four of the seven patients achieved scores within +/-1 standard deviation of the mean for normal-hearing listeners at +15 and +10 dB SNR. At the +5 dB SNR three patients achieved scores within +/-1 standard deviation of the mean for the normal-hearing listeners. For the HINT sentences, four of seven patients achieved scores within +/-1 standard deviation of the mean for the normal-hearing listeners at +15 dB and at +10 dB SNR and two achieved scores within that range at +5 dB SNR. CONCLUSION: Our results extend the range of stimulus conditions, from quiet to modest amounts of noise, in which the CIS strategy allows the best performing patients to extract most, if not all, of the information available to normal hearing subjects listening to speech processed into six channels. PMID- 9867297 TI - Consumer is not king in the health-care marketplace. PMID- 9867298 TI - A comparison of the cost effectiveness of one-stage versus two-stage bilateral total hip replacement. AB - Forty patients underwent 80 bilateral primary total hip replacements (THRs) under the same anesthesia (one-stage). Forty other patients who underwent unilateral primary THRs during the same time interval were selected to match the first 40 patients with regard to age, sex, diagnosis, weight, medical comorbidity, type of prosthesis used, and perioperative management protocol. An assumption was made in that each unilateral case represented the first side of bilateral THRs performed during two separate hospitalizations (two-stage). Analysis of the total hospital charges submitted to the insurance companies was made between the groups. On average, there was a 24% reduction (P<.05) for each case if bilateral THRs were done in one stage. This was primarily due to a significant decrease (P<.05) in the length of hospital stay in the one-stage group. There was no difference between the two groups in the operative time, estimated blood loss, or perioperative complications. PMID- 9867299 TI - Progressive opening-wedge osteotomy for severe tibia vara in adults. AB - Five patients with tibia vara were treated with progressive opening-wedge osteotomy. The maximum varus deviation of the mechanical axis of the tibia varied from 20 degrees to 12 degrees. The minimum postoperative follow-up was 26 months. Complete correction of the deformity was achieved without any major complications. Progressive opening-wedge osteotomy offers several advantages over conventional osteotomy in adults. First, fibular osteotomy is unnecessary. Second, knee mobility is only slightly restricted immediately following surgery. Third, it is possible to adjust correction postoperatively, and finally, progressive opening-wedge osteotomy does not induce shortening of the lower limb. The most significant disadvantage is the need for prolonged external fixation. This technique should be reserved for patients with severe deformities (minimum of 15 degrees varus) and mild or moderate osteoarthrosis. PMID- 9867300 TI - Treatment of fibrous dysplasia involving the proximal femur. AB - Twenty-two patients with fibrous dysplasia in the femoral neck or trochanteric area were treated with curettage and bone grafting with a sliding hip compression screw and plate. Follow-up ranged from 2 to 6 years (average: 4 years). Fourteen patients had monostotic and 8 had polyostotic disease. Four patients had pathologic fractures. Bone grafting included a deep-frozen allogeneic cortical strut and cancellous bone. After implanting the lag screw and cortical strut, the remaining defect space was filled with iliac bone. Postoperatively, all patients had good bone healing and complete incorporation of the implanted graft. There were no recurrences or complications, and functional results were rated as good and excellent. PMID- 9867301 TI - Fatal pulmonary embolism after total hip arthroplasty: diurnal variations. AB - From 1970 through 1986, a total of 18,104 Charnley low-friction arthroplasties were performed; of these, 122 deaths occurred from pulmonary embolism within 1 year of surgery. Diagnosis was confirmed by postmortem examination in 71% of cases. The exact time of the onset of the complication was recorded in 90 cases. In 74 (82%) cases, the time of collapse occurred during the 7-hour period from 9:00 AM to 4:00 PM, and in 16 (18%) cases, it occurred in the 17-hour period from 4:00 PM to 9:00 AM. The patient's activity at the time of collapse was recorded in 73 cases. Sixty (82%) were mobile, 3 were in the bathroom, and 10 (14%) were in bed. Sixty-six (70.2%) patients died within 1 hour of the onset of symptoms. PMID- 9867302 TI - Prevention of venous thrombosis after elective hip surgery. PMID- 9867303 TI - Anterior endoscopic correction and fusion of scoliosis. PMID- 9867304 TI - Morton's neuroma following first metatarsal osteotomy. PMID- 9867305 TI - Cervical spondylitis and epidural abscess caused by Salmonella enteritidis with tetraplegia. PMID- 9867306 TI - Slipped capital femoral epiphysis in identical twins: HLA predisposition. PMID- 9867308 TI - The British National Health Service (NHS) celebrates its 50th anniversary: equity in a competitive world. PMID- 9867307 TI - Rupture of the ulnar collateral ligament of the first metacarpophalangeal joint ("gamekeeper's thumb") PMID- 9867309 TI - The glutamate hypothesis of schizophrenia. PMID- 9867310 TI - In vivo age-related changes in hepatic drug-oxidizing capacity in humans. AB - Very few studies have been carried out looking at how the effects of drugs and their toxicity in humans change during their lifespan (developing and ageing). The purpose of this study is to review the literature on the changes in probe drug metabolism, classified by cytochrome P450 (P450 or CYP) at five stages in life: neonates < 4 weeks, infants < 12 months, children < 19 years, young/mature adults 20-64 years, and elderly adults > 65 years. The main probe drugs include caffeine and theophylline, whose metabolism is catalysed by CYP1A2, tolbutamide, phenytoin and ibuprofen, catalysed by CYP2C9, amitriptyline and nortriptyline, catalysed by CYP2C19, acetaminophen, catalysed by CYP2E1 and lidocaine, midazolam and terfenadine, catalysed by 3A3/4. From the published in vivo studies two different patterns of drug metabolism can be identified: (i) activity is low immediately after birth, increases, then peaks at the young/mature adult level and, finally, decreases in old age (drugs catalysed by CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A3/4) and (ii) activity increases rapidly after birth to reach a level equivalent to that in the young/mature adult, then gradually decreases and finally decreasing faster in old age (drugs catalysed by CYP2E1). Further study of the changes in P450 with age is warranted to help prevent adverse reactions and to guide us in tailoring therapy better for the individual patient. PMID- 9867311 TI - Design and delivery of non-parenteral vaccines. AB - Non-parenteral delivery of vaccines is reviewed focusing on the delivery systems that have been used for various mucosal routes of administration. Systems considered include biodegradable micro- and nanoparticles, liposomes, live bacterial and viral vectors and mucosal adjuvants. New approaches to mucosal vaccine formulation using: (i) gene fusion technology to create non-toxic derivatives of mucosal adjuvants, (ii) genetically inactivated antigens with a deletion in an essential gene, (iii) coexpression of an antigen and a specific cytokine that is important in the modulation and control of a mucosal immune response, and (iv) genetic material itself that would allow DNA or RNA uptake and its endogenous expression in the host cell are described. PMID- 9867312 TI - Drug utilization in a hospital general medical outpatient clinic with particular reference to antihypertensive and antidiabetic drugs. AB - OBJECTIVES: To examine the prescribing patterns, particularly antihypertensive and antidiabetic drugs, in a hospital outpatient clinic and to evaluate the expenditure incurred. METHODS: Prescriptions from a general medical outpatient clinic in a teaching hospital were collected for 4 weeks. Drug expenditures were calculated. RESULTS: A total of 622 prescriptions containing 1903 items were collected. Antihypertensive and antidiabetic drugs accounted for 25.9 and 9.1% usage, respectively. Calcium channel blocking agents, angiotensin converting enzyme inhibitors and beta-blocking agents were the most popular antihypertensive drugs used. Sulphonylureas were the most frequently prescribed antidiabetic agents. Amlodipine, doxazosin and sotalol accounted for 4.5% of all prescription items but 27.8% of the total drug expenditure. CONCLUSION: The use of antihypertensive and antidiabetic drugs accounted for most of the drug usage in this study. Use of newer and more expensive agents has further increased drug expenditure. Monitoring drug usage and their correlations with clinical outcomes are warranted. PMID- 9867313 TI - Use of indapamide in hospital and community clinics and its effect on plasma potassium in Chinese patients. AB - OBJECTIVES: To investigate the usage pattern of indapamide and other antihypertensive drugs in patients attending a community-based government outpatient clinic (GOPC) or a hospital-based specialist clinic (SC). The plasma potassium concentrations of patients receiving indapamide and other diuretics were also examined. METHOD: Prescriptions from the SC and the GOPC were reviewed and collected during January 1998. Patients' plasma potassium concentrations and the date of initiation of each medication were retrieved from the hospital computer databases at SC. An age- and sex-matched control group of patients on non-diuretic antihypertensive drugs was identified. RESULTS: A total of 1648 and 773 prescriptions were collected from the SC during a 1-week period and GOPC during a 1-month period, respectively. Approximately half (45%) of the patients received antihypertensive treatment. Indapamide was five times more frequently prescribed in GOPC than SC (84.7 vs. 17.7%, P<0.001). Calcium channel blocking agents were the commonest antihypertensive drugs used in both clinics. The mean plasma potassium concentration of patients taking indapamide was lower than that of the control group (P = 0.037). Multiple linear regression analysis showed that consumption of indapamide (P =0.002) and duration of diuretic therapy (P = 0.023) were significantly related to changes in plasma potassium concentrations [multiple regression equation for potassium level = 4.09-0.145 (thiazide = 1) 0.377 (indapamide = 1) -0.00468 (duration of diuretic therapy in months)]. CONCLUSION: Indapamide was used extensively in the community clinic and less in the hospital-based outpatient clinic. Patients receiving indapamide had a significantly lower plasma potassium concentration as compared to other diuretics or antihypertensive groups and this was predicted by a multiple linear regression model. Monitoring plasma electrolytes before initiation of indapamide treatment and at regular intervals thereafter is essential for detecting the hypokalaemia that may occur in Chinese patients. PMID- 9867314 TI - Decrease in emergency room or urgent care visits due to management of bronchial asthma inpatients and outpatients with pharmaceutical services. AB - OBJECTIVE: To evaluate pharmaceutical care of bronchial asthma over a 1-year period and to investigate whether such care is effective. METHODS: 79 male and 97 female asthma patients who were treated with theophylline and other drugs between June 1994 and September 1996 were studied. The 15 subjects received pharmaceutical care (group 1) and were all inpatient asthmatics who had been admitted to the hospital for treatment. The other patients acted as controls. RESULTS: In 1994, the average medication cost of asthma patients without pharmaceutical care was $199/month and $295/month when such care was provided in the outpatient clinic. The drug treatments which patients received generally included inhaled anti-inflammatory agents and theophylline, but the 161 non pharmaceutical services patients (group 2) received fewer inhaled anti inflammatory agents than group 1 patients. The number of visits to the emergency room and hospital admissions were significantly lower in patients after receiving pharmaceutical services than inpatients before receiving pharmaceutical services during our observation period. The serum theophylline concentrations of group 1 patients generally attained levels sufficient for favourable therapeutic effect after patients received pharmaceutical care. CONCLUSION: Pharmaceutical care offered at the hospital is effective in the management of asthma patients. PMID- 9867315 TI - Determination of compatibility and stability of drugs used in palliative care. AB - BACKGROUND: Drugs for symptom control in the terminal phase of palliative patients may be used in pump systems. OBJECTIVE: To investigate the compatibility and stability of solutions of morphine hydrochloride, haloperidol, metoclopramide hydrochloride, atropine sulphate, butylhyoscine bromide and ranitidine hydrochloride, which may be used together under such circumstances. METHOD: Solutions of the drugs were exposed to ambient light at a temperature of 20 +/- 5 degrees C and +/- 31 degrees C for 24 h and 7 days and the solutions studied by ultraviolet spectrophotometry. RESULTS: The combination of haloperidol or metoclopramide hydrochloride with different concentrations of morphine hydrochloride seemed to be compatible and stable. The concentration of atropine sulphate or butylhyoscine bromide could not be determined by the study method, but morphine hydrochloride seemed to be stable. The combination of ranitidine hydrochloride and morphine hydrochloride showed a change in colour after 7 days. CONCLUSION: The results suggest that the combination of different concentrations of morphine hydrochloride with haloperidol, metoclopramide hydrochloride, atropine sulphate, butylhyoscine bromide or ranitidine hydrochloride do not affect their stability when stored for 7 days up to 30 degrees C under the influence of ambient light. However, more robust stability-indicating methods are required to confirm these results. The proposed method is more useful for identifying combinations that are clearly incompatible than to identify those that are compatible. PMID- 9867316 TI - Kinetic profile of carbamazepine in an adult Portuguese outpatient population. AB - OBJECTIVE: The aim of our work was to define the kinetic profile of carbamazepine (CBZ), in order to improve on dosing schedules through a Bayesian approach. METHOD: Carbamazepine dose/steady-state trough concentrations data pairs and associated information were collected retrospectively on a population of adult epileptic patients. RESULTS: Fifty patients (index population) with two or more available concentrations (total of 174 determinations) met our inclusion criteria. Patients were taking CBZ (200-1800 mg/day) in mono- or polytherapy regimens. The analysis assumed a one-compartmental model with first-order absorption and elimination. Due to the data source (only trough concentrations were measured as part of hospital routine), the volume of distribution was fixed at 1.19 l/kg. The final estimates for CL were: 0.075 +/- 0.027 (mono- and polytherapy), 0.069 +/- 0.020 (monotherapy), and 0.106 +/- 0.037 l/h/kg (polytherapy). In order to validate these results, we assessed their predictive capacity using 18 new patients (validation population), submitted to the same inclusion criteria and using Prediction-Error analysis. The results suggested a different CL value for our population compared to earlier published clearance values. The results also pointed to an increased metabolic rate associated with polytherapy. The prediction capacity of the optimization method derived from a Portuguese population made in an a priori evaluation indicated a low error (-0.04 microg/ml), close to the theoretical zero value. CONCLUSION: Our results provide specific data on CBZ disposition in a Portuguese population and given the wide variability in the literature values, our data may help improve dosing of CBZ in Portuguese patients. PMID- 9867318 TI - Type III and type IV endoleak: toward a complete definition of blood flow in the sac after endoluminal AAA repair. AB - In this document the authors continue to refine their seminal categorization of endoleak, a major complication of endovascular aneurysm repair. In addition to type I (related to the graft device itself) and type II (retrograde flow from collateral branches) endoleak, they propose two new categories: endoleak due to fabric tears, graft disconnection, or disintegration would be classified type III, and flow through the graft presumed to be associated with graft wall "porosity" would be categorized as type IV endoleak. PMID- 9867317 TI - Angioplasty and stenting of the extracranial carotid arteries. AB - PURPOSE: To study the feasibility and safety of stent-supported angioplasty in the treatment of atherosclerotic stenoses of the extracranial carotid arteries. METHODS: Carotid angioplasty was attempted in 174 arteries (163 patients: 126 males; mean age: 71 +/- 10 years, range 47 to 93). Mean lesion length was 15.1 +/ 4.1 mm, and mean percent stenosis was 83.8% +/- 7.3% (reference diameter 5.8 +/- 0.7 mm). The majority (106, 65%) were asymptomatic (51% of all patients had severe coronary disease, 32% had peripheral vascular diseases). Patients underwent independent neurological examination, computed tomography, duplex ultrasonography, and angiography preprocedurally, 24 hours after the procedure, and at 6-month follow-up intervals. Most (142, 82%) carotid arteries were treated without cerebral protection, but a protective triple coaxial catheter was used in 32 (18%) patients. Stents (primarily Palmaz and Wallstent) were deployed routinely in all cases; 18% were implanted without predilation. RESULTS: Immediate technical success was 173 of 174 (99.4%) (1 access failure referred electively to surgery). Eight (4.6%) neurological complications occurred in the periprocedural period: 3 transient ischemic attacks, 2 minor strokes, and 3 major strokes (1 amaurosis and 2 hemiplegias). Two major complications developed despite cerebral protection. There were no deaths or myocardial infarctions and only 3 cervical access site hematomas. Over a mean 12.7 +/- 9.2 month follow-up (range 1 to 36), no ipsilateral neurological complications have been seen. There were 4 (2.3%) restenoses (3 redilated, 1 treated medically) and 1 mild Palmaz stent compression, all found within the first 6 months. Primary and secondary patencies at 3 years are 96% and 99%, respectively. CONCLUSIONS: Angioplasty with routine stenting seems feasible and safe for treating certain types of carotid stenoses even in high-surgical risk patients; however, randomized trials are necessary before this treatment can be offered as an alternative to endarterectomy. PMID- 9867319 TI - Endovascular AAA exclusion: will stents with hooks and barbs prevent stent-graft migration? AB - PURPOSE: To investigate if stents with hooks and barbs will improve stent-graft fixation in the abdominal aorta. METHODS: Sixteen- to 24-mm-diameter Dacron grafts were deployed inside cadaveric aortas. The grafts were anchored by stents as in endovascular abdominal aortic aneurysm repair. One hundred thirty-seven stent-graft deployments were carried out with modified self-expanding Z-stents with (A) no hooks and barbs (n = 75), (B) 4 5-mm-long hooks and barbs (n = 39), (C) 8 10-mm-long, strengthened hooks and barbs (n = 19), or (D) hooks only (n = 4). Increasing longitudinal traction was applied to determine the displacement force needed to extract the stent-grafts. The radial force of the stents was measured and correlated to the displacement force. RESULTS: The median (interquartile range) displacement force needed to extract grafts anchored by stent A was 2.5 N (2.0 to 3.4), stent B 7.8 N (7.4 to 10.8), and stent C 22.5 N (17.1 to 27.9), p < 0.001. Both hooks and barbs added anchoring strength. During traction, the weaker barbs were distorted or caused intimal tears. The stronger barbs engaged the entire aortic wall. The radial force of the stents had no impact on fixation, while aortic calcification and graft oversizing had marginal effects. CONCLUSIONS: Stent barbs and hooks increased the fixation of stent grafts tenfold, while the radial force of stents had no impact. These data may prove important in future endograft development to prevent stent-graft migration after aneurysm exclusion. PMID- 9867320 TI - Renal complications following endovascular repair of abdominal aortic aneurysms. AB - PURPOSE: To investigate the renal complications associated with endovascular repair of abdominal aortic aneurysms (AAAs). METHODS: Data were prospectively collected on 164 AAA patients (154 males; median age 72 years; interquartile range 51 to 88) undergoing endovascular grafting. Any history of renal failure and diabetes mellitus was recorded. Serum urea and creatinine levels were measured preoperatively and at regular intervals postoperatively. Renal impairment was defined as serum creatinine > 130 micromol/L. RESULTS: There were no significant differences in pre- and 1-day postoperative serum urea and creatinine levels. Among 15 (9.1%) patients with preoperative renal failure, 7 (47%) died, 4 (27%) in the perioperative period. Of the 149 patients with normal renal function preoperatively, 4 (2.7%) developed renal failure as part of multisystem organ failure. Another 9 (6.2%) developed significant postoperative elevations (> 20%) in their creatinine levels compared to baseline; 4 of these patients died, 2 in the perioperative period. There was no significant difference in the median dose of intravascular contrast used for those patients that did and did not have a deterioration in their renal function (250 mL versus 300 mL). CONCLUSIONS: In this study, approximately 6% of patients with normal preoperative renal function who undergo endovascular AAA repair develop renal dysfunction. For patients with preoperative renal impairment, the perioperative mortality rate is high, 27%, following endovascular aortic aneurysm repair. PMID- 9867321 TI - Combined surgical and endovascular removal of thrombus entirely occluding a bifurcated aortic stent-graft. AB - PURPOSE: To explore a method combining interventional, endovascular, and conventional surgical techniques for treating a completely occluded bifurcated stent-graft after endovascular aortic aneurysm repair. METHODS AND RESULTS: A 60 year-old patient underwent endovascular repair of an abdominal aortic aneurysm (AAA) with a Talent bifurcated stent-graft. Five months later, after chronic thrombotic occlusion of the right iliac limb, he presented with acute occlusion of the entire stent-graft. Local intra-arterial infiltration thrombolysis successfully reconstituted flow through the main aortic segment and left iliac limb. With a combination of conventional surgical and intraoperative endovascular procedures, thrombectomy and recanalization of the right iliac limb was completed by stenting a severe stenosis of the proximal iliac limb. CONCLUSIONS: A combination of techniques may be essential for successful management of thrombotic complications after endovascular AAA repair. PMID- 9867322 TI - Combined approach to stent-graft treatment of an aortic arch aneurysm. AB - PURPOSE: To report the first use of dual brachiocephalic vessel transposition from the aortic arch prior to endovascular exclusion of an arch aneurysm. METHODS AND RESULTS: Patch graft repair of a saccular aortic arch aneurysm in a 74-year old woman failed to exclude the lesion, resulting in aneurysm enlargement. Because the patient was debilitated, a combined open and endovascular procedure was performed in which the common carotid and subclavian arteries were transposed prior to successful insertion of a Vanguard aortic stent-graft to exclude the aneurysm. Imaging 1 and 6 months after the combined procedure demonstrated complete thrombosis of the aneurysm. CONCLUSIONS: Transposition from the aortic arch of one or more brachiocephalic vessels may allow stent-graft exclusion of aortic arch aneurysms. However, the durability of thoracic aortic endografts must be ascertained before the indications for their use are expanded. PMID- 9867323 TI - Combining endovascular and surgical techniques: the best of both worlds. PMID- 9867324 TI - Laparoscopic aortic aneurysm resection. AB - PURPOSE: To describe a laparoscopic technique for resection of infrarenal abdominal aortic aneurysms (AAAs). METHODS: The operation is based on the principle of retroperitoneal reinforced staple exclusion of the aneurysm sac with aortobifemoral or aortoiliac bypass using gas and gasless laparoscopic techniques. Patients were eligible for this procedure if their infrarenal AAAs (with or without iliac artery involvement) were considered appropriate for surgical resection; however, renal or other visceral arterial stenoses, aneurysmal disease requiring surgical treatment, and/or aneurysms of the hypogastric arteries excluded patients from laparoscopic AAA resection. RESULTS: Of 31 candidates for this procedure, 9 were excluded owing to high surgical risk. Twenty-two patients (16 males; age range 62 to 88 years) were deemed appropriate for the laparoscopic procedure. Maximum aneurysm diameter ranged from 4.0 to 8.0 cm. The operation was completed successfully in 20 (91%) patients. Two (9%) deaths in high-risk patients admitted early to the study occurred within 30 days of surgery. The only major complication was an injured ureter, for which a nephrectomy was performed. Comparison to a historical cohort of conventionally treated patients showed that the study group needed less ventilator support, had shorter intensive care and hospital stays, and resumed diet earlier despite relatively prolonged anesthesia and aortic clamping times. CONCLUSIONS: The laparoscopic approach to infrarenal AAAs appears feasible, with several potential advantages in low- and moderate-risk patients. Once the technique is optimized, randomized prospective studies will be needed to verify the apparent benefits demonstrated by these initial patients. PMID- 9867325 TI - Endovascular treatment of internal iliac artery aneurysms. AB - PURPOSE: To describe a technique for percutaneous endovascular repair of internal iliac artery aneurysms. TECHNIQUE: In a patient with an isolated internal iliac artery aneurysm and combined tight stenoses at the origins of the external and internal iliac arteries, interventional treatment consisted of coil embolization of the internal iliac artery and branches using the contralateral femoral approach. Balloon dilation of the external iliac stenosis occluded the internal iliac artery, and Palmaz stents were placed in the external iliac stenosis to overcome residual luminal narrowing. CONCLUSIONS: This one-step percutaneous technique allows total exclusion of the internal iliac artery aneurysm using coil embolization. This treatment may be an alternative to surgery in isolated internal iliac artery aneurysms with small orifices. PMID- 9867326 TI - The effect of nonporous PTFE-covered stents on intimal hyperplasia following balloon arterial injury in minipigs. AB - PURPOSE: To report an experimental study investigating the ability of nonporous polytetrafluoroethylene (PTFE) covering on a metallic stent to retard the development of neointimal hyperplasia (NIH). METHODS: Three groups of Hanford miniature swine underwent standardized balloon injury to both external iliac arteries. Group I animals (control) received balloon injuries only. Group II had the site of balloon injury supported by a properly sized, balloon-expandable Palmaz stent placed directly over the injury site. Group III animals received a Palmaz stent covered with PTFE graft. All animals underwent arteriography immediately after intervention and again prior to sacrifice and specimen harvest at 4 weeks. The specimens were examined grossly and histologically at the proximal, middle, and distal segments for NIH development. RESULTS: Uncovered stents developed significantly more NIH (p < 0.0001) and greater luminal narrowing (p < 0.001) than the controls. PTFE-covered stents (group III) exhibited less NIH (p < 0.001) and luminal reduction (p < 0.01) than bare stents (group II) at the middle portion of the stent-graft, but the PTFE cover had no effect on NIH and lumen reduction at the proximal or distal ends of the prosthesis. CONCLUSIONS: PTFE-covered stents retarded NIH at 4 weeks, but only at the midportion of the devices; the covering did not prevent neointimal pannus ingrowth at the proximal and distal ends. PMID- 9867327 TI - In vivo crushing of an aortic stent enables endovascular repair of a large infrarenal aortic pseudoaneurysm. AB - PURPOSE: To report an unusual approach to endovascular exclusion of a large aortic pseudoaneurysm. METHODS AND RESULTS: A 63-year-old male had an unsuccessful endovascular repair of an aortic anastomotic pseudoaneurysm that left an expanded, uncovered Palmaz stent lying obliquely within the aorta. After nearly 3 years, the pseudoaneurysm enlarged to 7 cm, and the patient became symptomatic. Repair of the pseudoaneurysm was accomplished by crushing the indwelling stent to allow placement of a stent-graft. CONCLUSIONS: Malpositioned stents that are hindering an endoluminal procedure may be crushed against the arterial wall in vivo to facilitate passage of endovascular instruments or devices. PMID- 9867328 TI - Initial experiences with percutaneous endovascular repair of popliteal artery lesions using a new PTFE stent-graft. AB - PURPOSE: To report the percutaneous endovascular management of popliteal artery lesions with a new polytetrafluoroethylene (PTFE) stent-graft. METHODS AND RESULTS: Three patients have undergone endovascular treatment in the popliteal region owing to: (1) an iatrogenic false aneurysm with arteriovenous fistula, (2) entrapment syndrome, and (3) an isolated arterial aneurysm. In each case, arterial occlusion was treated initially with thrombolysis. The underlying vascular lesions then were stented using the new Hemobahn endograft, a nitinol stent covered internally with PTFE. Each percutaneous procedure was completed successfully without major complications. During early follow-up (6 to 7 months), no signs of intimal hyperplasia, occlusion, or stent migration have been observed. CONCLUSIONS: Based on these early experiences, percutaneous treatment of stenotic and aneurysmal lesions in the popliteal artery using the Hemobahn endograft appears feasible and effective. Further experience and longer follow-up are required to evaluate this new prosthesis. PMID- 9867329 TI - Successful endoluminal repair of a popliteal artery aneurysm using the Wallgraft endoprosthesis. AB - PURPOSE: To describe the first application of the Wallgraft endovascular prosthesis to aneurysm exclusion in the popliteal artery. METHODS AND RESULTS: A 61-year-old man was admitted to our institution with a popliteal artery aneurysm complicated by associated popliteal vein compression, deep venous thrombosis, and pulmonary embolism. The endovascular procedure was performed percutaneously with local anesthesia using a low-profile, covered stent (Wallgraft Endoprosthesis). The aneurysm was successfully excluded from the arterial circulation, and there were no postprocedural complications. Follow-up evaluation with appropriate imaging at 10 months revealed no endoleaks. CONCLUSIONS: This initial result indicates that percutaneous deployment of the Wallgraft endoprosthesis, with its smaller diameter and flexible design, may offer significant advantages over currently available devices for repair of popliteal artery aneurysms. This method obviates the need for general anesthesia or surgical exposure, which is particularly beneficial in patients with comorbid illnesses. PMID- 9867331 TI - A grading scale to predict the degree of difficulty for endovascular AAA graft procedures. PMID- 9867330 TI - "Retroleak"--retrograde branch filling of the excluded aneurysm. PMID- 9867332 TI - A novel method for controlling the quantity of mitomycin-C applied during filtering surgery for glaucoma. AB - The use of a sponge impregnated with mitomycin-C is an increasingly common practice in glaucoma surgery. The appropriate antibiotic concentration and exposure time have been considered in the literature, but not the exact amount to be used or the size of the sponge. The purpose of this study was to estimate the quantity of mitomycin-C contained in sponges prepared by different surgeons as compared to that in applicators of the type used in Schirmer's test graduation (5 x 5 mm). Four surgeons each cut and prepared 10 sponges for intraoperative use according to their usual method. The same procedure was performed with 10 Schirmer's test graduations. Each sponge and each graduation was immersed in a solution of mitomycin-C 0.2 mg/ml, and the quantity of antibiotic (microg) in each was calculated as the difference between wet and dry weight. The mean quantity (+/- SEM) of mitomycin-C contained in cut sponges was 9.6 +/- 4.4 microg (range 1.9-17.3), and the differences between surgeons were statistically significant (p<0.0001). The mean quantity of antibiotic in Schirmer's test graduation was 1.7 +/- 0.3 microg (range 1.1-2.5), and the differences between surgeons were not statistically significant (p=0.79). The quantities of mitomycin C contained in sponges prepared for glaucoma surgery differed for a given surgeon and between surgeons. Thus, variations in the doses applied to the sclera could account for certain complications due to mitomycin-C. The use of a Schirmer's test graduation improves predictability for the quantity of mitomycin-C applied to the sclera. PMID- 9867333 TI - The effect of a new tear substitute containing glycerol and hyaluronate on keratoconjunctivitis sicca. AB - The purpose of this study was to evaluate the efficacy of LO2A, a newly developed tear substitute containing glycerine and sodium hyaluronate, in the treatment of dry eyes. Twenty-five informed consent patients suffering from keratoconjunctivitis sicca were included. Patients were treated for one week with LO2A in one eye, and with their current tear substitute in the other eye. Rose bengal staining was evaluated on a scale of 0 to 3. Patient satisfaction was graded on a scale of 1 to 5. The average satisfaction score for LO2A was significantly higher compared to the control preparations at 1 week (p=0.0003) and at 2 weeks (p=0.0232). A highly significant reduction in rose bengal staining was demonstrated following 1 week of treatment with LO2A (p<0.0001). The LO2A treated eyes had significantly less staining than control eyes at 1 week (p=0.021) and at 2 weeks (p=0.023). An inverse correlation was found between patient grading and the rose bengal scoring (spearman rank coefficient = -0.49, p<0.001). LO2A showed a beneficiary effect on dry eye patient satisfaction and on rose bengal test, as compared to other tear substitute preparations currently used by these patients. PMID- 9867334 TI - N-linked glycoside and glucuronide conjugates of the retinoid, acitretin, are biologically active in cornea and conjunctiva. AB - The purpose of this study was to test two water-soluble, synthetic retinoids, glucoseamido acitretin and glucuronamido acitretin, for biological activity in cells of the cornea and conjunctiva. Vitamin A-deficient, xerophthalmic rats were treated topically with these retinoids, and corneas were examined histologically for effects on epithelial keratinization. The effect of these retinoids on the proliferation of rabbit conjunctival fibroblasts in culture was also investigated. Glucoseamido acitretin treatment restored a normal cornea after eight to nine days of treatment, while no improvement was observed in the vehicle treated corneas. Likewise, glucuronamido acitretin application restored a normal corneal surface and reversed keratinization after eight to ten days of treatment. These retinoids caused no irritation of the eye or ocular adnexa. In culture, exposure of conjunctival fibroblasts to glucoseamide acitretin inhibited cell proliferation. Cultures exposed to glucoseamido acitretin at 10(-8) M or 10(-6) M had cell densities 77.3% and 51.9% of control, respectively, after seven days. Glucuronamido acitretin also inhibited cell proliferation. Cultures exposed to glucuronamido acitretin at 10(-8) M had a cell density of 69.2% of control at day seven, while at 10(-6) M this retinoid completely inhibited cell proliferation. These results show that glucoseamide acitretin and glucuronamido acitretin are biologically active in the cornea and conjunctiva, and may be considered for ophthalmic use in diseases involving abnormalities of ocular surface cell differentiation or hyperproliferation of fibroblasts. PMID- 9867335 TI - Expression of ICAM-1 and HLA-DR by human conjunctival epithelial cultured cells and modulation by nedocromil sodium. AB - It is unclear whether conjunctival epithelial cells participate in the development of immune-mediated events. Using a previously reported in vitro system of human conjunctival epithelium, we determined whether conjunctival epithelial cells express two relevant markers in the antigenic presentation process. Moreover, the potential capability of nedocromil sodium, an antiallergic and antiinflammatory drug, to modulate such expression was investigated. Primary cultures of human conjunctival epithelium and Chang conjunctival cells, incubated with or without 100 U/ml IL-1beta and/or IFNgamma for 1, 3 or 6 h, were simultaneously exposed to 10(-5) M nedocromil sodium. The expression of the intercellular adhesion molecule-1 (ICAM-1) and the human leukocyte antigen-DR (HLA-DR) was determined immunocytochemically. Constitutive expression of ICAM-1 and HLA-DR was observed in primary cultures and Chang cells and was minimally affected by incubation with IL-1beta and/or IFNgamma. The addition of nedocromil sodium resulted in complete abolition of HLA-DR expression and a notable reduction in ICAM-1 expression in primary cultures and Chang cells. These results suggest that epithelial cells from human conjunctiva constitutively express ICAM 1 and HLA-DR in vitro and that such expression is downregulated by nedocromil sodium. This may indicate that conjunctival epithelial cells may be another target for this drug. PMID- 9867336 TI - The conjunctival provocation test model of ocular allergy: utility for assessment of an ocular corticosteroid, loteprednol etabonate. AB - Two studies were conducted using the conjunctival provocation test (CPT) model of ocular allergy. The objective of the first study was to evaluate the sensitivity of the CPT model to a topical corticosteroid. Selected was loteprednol etabonate 0.5%, previously found effective in the treatment of ocular allergy and inflammation. The study was a randomized double-masked, placebo-controlled, paired-comparison of loteprednol etabonate 0.5% (LE), b.i.d. or q.i.d. Sixty subjects who had a minimum pre-determined allergic response received LE in one eye and placebo in the fellow eye for 28 days from Day 7 to Day 35. Antigen challenges were carried out on Days 0, 7 (baseline), 21 and 35. The primary endpoints were interocular differences in itching and mean redness (the average of ciliary, conjunctival and episcleral vessel beds). LE (either b.i.d. or q.i.d.) was significantly more effective than placebo for reducing mean redness and itching. No clinical or statistically significant changes in intraocular pressure were observed. Based upon the results of Study 1, we used the CPT model to aid in the selection of a concentration of loteprednol etabonate for subsequent studies in environmental seasonal allergic conjunctivitis. This was a randomized double-masked, placebo-controlled, paired-comparison of loteprednol etabonate 0.1%, 0.2% and 0.3%, q.i.d. in 88 subjects. The dosing and testing regimen was similar to the first portion of the study. Loteprednol etabonate, 0.1%, 0.2% and 0.3%, was numerically superior to the placebo in reducing mean redness and itching. At the 20-minute post allergen challenge, the 0.1% concentration was significantly superior (p < 0.05) to the placebo on Visit 4 (2 and 4 hour challenge) in reducing the mean redness; however, LE was only numerically superior in relieving itching. The 0.2% concentration was significantly superior (p < 0.05) to the placebo in the reduction of mean redness and itching on Visit 3 (Day 21) and in reduction of mean redness on Visit 4 (4 hour challenge). The 0.3% concentration was significantly superior (p < 0.05) to the placebo in the reduction of mean redness on all visits, and statistically significant in the reduction of itching on Visit 4 (4 hour challenge). While there were some elevations of IOP with LE 0.2%, they were not clinically significant. In conclusion, the CPT model of ocular allergy is useful in the evaluation of corticosteroids. Furthermore, based upon a dose-response study in this model, 0.2% loteprednol etabonate was selected for further evaluation in environmental seasonal allergic conjunctivitis studies. PMID- 9867337 TI - Binding of CGRP analogs and their effect on adenylate cyclase activity in porcine iris-ciliary body. AB - The structure-activity relationship of the calcitonin gene-related peptide (CGRP) in the porcine iris-ciliary body was studied using different CGRP analogs. The receptor binding affinity is located mainly in the carboxyterminal end of the CGRP peptide while the ability to stimulate adenylate cyclase (AC) enzyme is mainly in the aminoterminal end of the peptide. The binding of CGRP analogs was also found to be temperature-dependent. Changes in the alpha-helical region or in the beta-turn, as well as replacements of threonine-4, asparagine-25 or asparagine-26, reduce the binding affinity already at +4 degrees C. Truncated aminoterminus, changes in the loop region between cysteines 2 and 7, and especially in threonine 6, have for their part an important role in maintaining AC-stimulating activity. PMID- 9867338 TI - Pharmacokinetics of intravitreal vancomycin in normal and infected rabbit eyes. AB - The purpose of this study was to determine the pharmacokinetics governing the distribution and elimination of intravitreally injected vancomycin in normal and infected rabbit eyes. Two groups each of 36 pigmented animals were used. Group 1 served as control. In Group 2, experimental endophthalmitis was induced in the right vitreous by inoculation with Staphylococcus aureus. Once endophthalmitis developed, a vancomycin solution was injected. Four animals from each group were killed at nine time points post-injection, the vitreous and aqueous were removed, and blood samples were taken for HPLC analysis. Data analysis was performed using the RSTRIP program. The half-lives were 69 hours in normal vitreous and 14.53 hours in infected vitreous. Therapeutic drug levels were present in the vitreous 84 hours post-injection in all eyes; they were detected from 2 to 48 hours in normal aqueous but at lower levels in the infected ones. Kv and Ca/Cv ratios suggested that the primary route of elimination was across the retina and the anterior chamber in normal eyes, and via the retina in infected eyes. Results indicate that pharmacokinetic parameters change in pathological conditions, which may help establish better treatment guidelines for endophthalmitis. PMID- 9867339 TI - The ocular distribution of methazolamide after corneal and scleral administration: the effect of ionization state. AB - The accession of methazolamide in ionized and unionized form to cornea, sclera, aqueous humor and ciliary process was studied 10 minutes following separate application to either sclera or cornea of a 1 mM solution. Cornea and ciliary process concentrations were 27.3 and 14.5 microM for unionized application to cornea and 10.1 and 7.1 microM for ionized application. Bulk aqueous humor concentrations were much lower, 3.1 and 1.1 microM, and cannot account for drug found in ciliary process on this time scale. Scleral application of drug, by contrast, gave undetectable ciliary process concentrations. These results are presented as a model for drug disposition following single drop topical sulfonamide therapy. The scleral pathway for drug delivery to ciliary process was further tested by application to sclera of 300 microL of either a 1 mM ionized or unionized solution for 30 minutes or as a 2% (85.5 mM) ionized solution for 30 minutes. In these series, red cell carbonic anhydrase was presaturated at -24 hours with drug to remove a possible route for loss of drug from sclera, that of the systemic circulation and the high concentration of carbonic anhydrase in red cells. After either ionized or unionized application, approximately 1 microM was detected in ciliary process, but all drug was attributable to the blood content of the tissue and the drug bound to red cell carbonic anhydrase. After 2% dosing, 4 microM was detected in ciliary process after allowance for drug in red cells. This concentration is below that necessary for inhibition of ciliary process carbonic anhydrase, suggesting that especially with regard to topical sulfonamide therapy, the corneal route of drug delivery to ciliary process predominates greatly over the scleral route. PMID- 9867341 TI - Selected papers from the 1st meeting of the European Society for Reproductive and Developmental Immunology. Amsterdam, The Netherlands, June 26, 1997. PMID- 9867340 TI - Ocular penetration of oral clarithromycin in humans. AB - Clarithromycin has a wide spectrum of activity against many gram-positive and gram-negative organisms, intracellular pathogens, and opportunistic pathogens. To examine the penetration of clarithromycin in the ocular tissues, 21 patients who underwent elective cataract surgery (Group I) received a single 500-mg dose of clarithromycin orally either 4, 8, 10, 12, or 22 hours before cataract surgery, and 21 patients who underwent elective retina/vitreous surgery (Group II) received 500 mg every 12 hours orally for 3 days before the surgery with the last dose given either 3, 6, 8, 11, or 24 hours before the surgery. Serum from all patients was assayed for clarithromycin prior to drug administration and at the time ocular specimen was taken. Aqueous, iris, and vitreous samples were also assayed for clarithromycin concentration. The concentrations of clarithromycin in the aqueous fluid 4, 8, 10, 12, and 22 hours after administration were: (mean +/- SD) 0.13+/-0.05, 0.137+/-0.11, 0.074+/-0.03, 0.06+/-0.02, and 0.074+/-0.04 microg/ml, respectively. Concentration of clarithromycin in vitreous 3, 6, 8, 11, and 24 hours after administration were: (mean +/- SD) 0.11+/-0.02, 0.257+/-0.13, 0.27+/-0.21, 0.307+/-0.26 and 0.108+/-0.07 microg/ml, respectively. The mean concentration of clarithromycin in the iris was 6.2 microg/g. In conclusion, this data suggest that clarithromycin widely penetrates and adequately concentrates in the aqueous humor, vitreous humor, and iris tissue after oral administration and therefore is effective in the management of many infectious ocular conditions. PMID- 9867342 TI - Society for the Study of Behavioural Phenotypes 7th annual meeting. Cambridge, United Kingdom, November 13-14, 1997. Abstracts. PMID- 9867343 TI - Biomolecular Interaction, Recognition and Dynamics. Proceedings of a workshop. Barcelona, Spain, October 23-25, 1997. PMID- 9867344 TI - Computerized evaluation procedure for comparing the electrophoretic protein patterns of bacterial strains. AB - The advantages and drawbacks of different methods of evaluating electrophoretic (SDS-PAGE) protein patterns were studied by comparing Erysipelothrix rhusiopathiae strains. The computerized evaluation procedure recommended by the authors renders it possible to determine objectively the degree of similarity of the protein composition of the different strains according to molecular mass. The degree of similarity obtained in four parallel analyses of a given strain was 988 +/- 0.55 on a scale on which 1000 equals complete identity. the method can be used without adjustment even in those cases where the two strains to be compared contain unequal amounts of electrophoresed proteins. PMID- 9867345 TI - Anaerobic dechlorination of perchloroethene in an extractive membrane bioreactor. AB - An extractive membrane bioreactor (EMB) is described that used an undefined anaerobic culture to dechlorinate tetrachloroethene (C2Cl4) reductively in a synthetic wastewater. Comparable reactors described in the literature use set-ups where the bacteria are in direct contact with the wastewater, and thus would require the addition of significant quantities of nutrients to the wastewater stream in practical application. In the EMB, a silicone rubber membrane separates the microbial culture from the wastewater stream, so that addition of nutrients can be minimised. The EMB was operated continuously for 48 days and dechlorinated 359 micromol C2Cl4 (1 biomedium -1 day -1) on average. Lactate was fed as an electron donor and C2Cl4 dechlorination was verified by chloride measurements. Particular attention was paid to the reduction of transmembrane C2Cl4 flux caused by a membrane-attached biofilm. Following a start-up period, the reactor operation was stable and remained largely unaffected by biofilm thickness and oxygen contamination from the wastewater. PMID- 9867346 TI - Malignant melanoma: current controversies, future perspectives. Proceedings of a symposium. Barcelona, Spain, 27-28 February 1997. PMID- 9867347 TI - [Vth Meeting of the National Group for the Study of the Pancreas. Alicante, 23-24 October 1998. Abstracts]. PMID- 9867348 TI - Bithalamic involvement predicts poor outcome among children with thalamic glial tumors. AB - Clinical features and treatment of 36 consecutive pediatric patients with thalamic glial tumors confirmed by histology and characterized by neuroimaging were reviewed to identify prognostic factors. The median age at diagnosis was 10 years (range 1-18 years). Twenty-four patients had low-grade tumors (juvenile pilocytic astrocytoma n = 9, fibrillary astrocytoma n = 6, astrocytomas not otherwise specified n = 6, ganglioglioma n = 2 and oligodendroglioma n = 1) and 12 patients had high-grade tumors (glioblastoma multiforme n = 7, anaplastic astrocytoma n = 4 and unclassified malignant tumor n = 1). With a median follow up of 4.3 years among survivors, estimates of 4-year progression-free survival (PFS) and overall survival (OS) for the entire group are 28+/-10 and 37 +/- 10%, respectively. Low-grade tumors were associated with a significantly better 4-year PFS (36 +/- 12 vs. 0% for the high-grade group; p = 0.03) and OS (52 +/- 12 vs. 0%; p < 0.001). This review identified that bithalamic involvement, characterized by neuroimaging, exerted an independent and significant negative impact on PFS and OS for patients with low-grade tumors. Estimates of 4-year PFS and OS among patients with tow-grade bithalamic versus monothalamic tumors were 58 +/- 15 vs. 0% and 85 +/- 11 vs. 0% (p < 0.00001), respectively. The presence of bithalamic involvement did not affect outcome among patients with high-grade tumors. Additionally, age at diagnosis, enhancement with neuroimaging contrast, extension beyond the thalamus and extent of surgical resection did not correlate with overall outcome. Because treatment approaches varied during the study period, the impact of radiation therapy or chemotherapy could not be assessed. This contemporary, single-institution series of pediatric thalamic glial tumors demonstrates, for the First time, the statistical significance of bithalamic involvement as a marker of poor prognosis among patients with low-grade glial lesions. PMID- 9867349 TI - Molecular cloning and expression of two novel zebrafish semaphorins. AB - The large, conserved semaphorin/collapsin gene family encodes putative axon guidance molecules. We describe the cloning and expression of two n ovel zebrafish semaphorins that represent an increase in the size and diversity of the family. These semaphorins are expressed in unique and dynamic patterns during development. PMID- 9867350 TI - Expression of a cysteine-rich protein (CRP) encoding gene during early development of the trout. AB - Members of the cysteine-rich protein (CRP) define a subclass of LIM-only proteins implicated mainly in muscle differentiation. Until now, very little is known concerning the expression of CRP encoding genes during vertebrate development. We describe here the isolation of a trout (Oncorhynchus mykiss) gene encoding a cysteine-rich protein (TCRP) and the pattern of its mRNA accumulation during embryogenesis, focusing on somitogenesis. TCRP encodes a putative protein with two LIM domains linked to a short glycine-rich region that displays 86%, 76%, 67% identity with chicken CRP2, CRP1 and MLP/CRP3 proteins, respectively. Whole-mount in situ hybridisation showed that TCRP transcript is first detected just before somitogenesis in the paraxial mesoderm, while it is absent in the axial structures. During somitogenesis, the expression of TCRP was observed caudally in the elongating presomitic mesoderm and in the last formed somites. The labelling for TCRP was found to fade as the somites mature. At the end of the somitogenesis, TCRP transcripts accumulation was restricted to pronephros and bronchial arches. PMID- 9867351 TI - DAP-2, the Drosophila homolog of transcription factor AP-2. AB - Transcription factor AP-2 is essential for craniofacial, nervous system, and limb development in the mouse. We report here the cloning and expression pattern analysis of DAP-2, the Drosophila homolog of AP-2 family genes. DAP-2 is expressed in discrete regions of procephalic neuroectoderm, the brain, ventral nerve cord, and maxillary segment during Drosophila embryogenesis, and in the brain, optic lobes, ventral nerve cord, antenno-maxillary complex, and antennal and leg imaginal disks in third instar larvae. Protein sequence conservation and parallels between the embryonic expression patterns of DAP-2 and mammal ian AP-2 family genes indicate that transcription factor AP-2 has been structurally and functionally conserved during metazoan evolution. PMID- 9867352 TI - Generating green fluorescent mice by germline transmission of green fluorescent ES cells. AB - Green fluorescent protein (GFP) and its variants currently represent the only non invasive markers available for labeling mammalian cells in culture or in a multicellular organism through transgenesis. To date this marker gene has been widely used in the study of many organisms, but as yet has not found large-scale application in mammals due to problems encountered with weak fluorescence and instability of the wild-type protein at higher temperatures. Recently, though, several mutants have been made in the wild-type (wt) GFP so as to improve its thermostability and fluorescence. EGFP (enhanced GFP) is one such wtGFP variant. As a first step in assessing the use of EGFP in ES cell-mediated strategies, we have established a mouse embryonic stem (ES) cell lines expressing EGFP, which can be propagated in culture, reintroduced into mice. or induced to differentiate in vitro, while still maintaining ubiquitous EGFP expression. From the results presented we can suggest that: 1) possible improvements in the efficiency of transgenic regimes requiring the germline transmission of ES cells by aggregation chimeras can be made by the preselection chimeric embryos at the blastocyst stage: (2) the expression of a noninvasive marker, driven by a promoter that is active during early postimplantation development, allows access to embryos during a window of embryonic development that has previously been difficult to investigate (3) the behavior of mutant ES cells can be followed with simple microscopic observation of chimeric embryos or adult animals comprising green fluorescent cells/tissues. and (4) intercrosses of F1 mice and subsequent generations of animals show that progeny can be genotyped by UV light, such that mice homozygous for the transgene can be distinguished from hemizygotes due to their increased fluorescence. PMID- 9867353 TI - Papers from the 10th International Symposium on Pediatric Surgical Research. Zurich, Switzerland, October 16-17, 1997. PMID- 9867354 TI - DNA topoisomerases. PMID- 9867355 TI - Joint Meeting of the British Microcirculation Society and the Northern Vascular Biology Group. Manchester, United Kingdom, March 30-13, 1998. Abstracts. PMID- 9867356 TI - A first mutation in the human tissue factor pathway inhibitor gene encoding [P151L]TFPI. PMID- 9867357 TI - Semiquantitative Epstein-Barr virus polymerase chain reaction analysis of peripheral blood from organ transplant patients and risk for the development of lymphoproliferative disease. PMID- 9867359 TI - Auditing surgical outcome. 10 Years with the Swedish Vascular Registry--Swedvasc. PMID- 9867358 TI - Significance of enhanced cytokine receptor expression by glucocorticoids. PMID- 9867360 TI - Expression vectors and delivery systems. Web alert. PMID- 9867361 TI - Spontaneous repigmentation of vitiligo patches distant from the autologous skin graft sites: a remote reverse Koebner's phenomenon? PMID- 9867362 TI - Acute generalized exanthematous pustulosis induced by diltiazem: value of patch testing. PMID- 9867363 TI - Pityriasis rosea and herpesvirus 7: action or interaction? PMID- 9867364 TI - Yellow nail pigmentation due to methotrexate. PMID- 9867365 TI - [Fli-1 (Friend leukemia integration 1). Oncogene]. PMID- 9867366 TI - [Rb1 (Retinoblastoma). Tumor suppressor gene]. PMID- 9867367 TI - [SMAD4/DPC4 (small mothers against decapentaplegic deleted in pancreatic carcinoma, locus 4). Tumor suppressor gene]. PMID- 9867368 TI - [BRCA1 (breast cancer type 1). Tumor suppressor gene]. PMID- 9867369 TI - Proceedings of the 3rd International Meeting on Advances in Perinatal and Paediatric Nutrition. Manchester, England, September 1997. PMID- 9867370 TI - 13th International Bone Densitometry Workshop. Delavan, Wisconsin, USA. 4-8 October 1998. Abstracts. PMID- 9867371 TI - 1997 Van Meter Award to Peter R. Arvan. PMID- 9867372 TI - 1997 Paul Starr Award to John T. Dunn. PMID- 9867373 TI - 1997 Sidney H. Ingbar Distinguished Lecturer Award to Jerome M. Hershman. PMID- 9867374 TI - Protecting children from armed conflict. Are most casualties non-combatants? PMID- 9867375 TI - Protecting children from armed conflict. Repaying debts takes precedence over health care in many Third World countries. PMID- 9867376 TI - Attention deficit hyperactivity disorder in children. Benefits of adding other forms of treatment to medication remain unclear. PMID- 9867377 TI - Attention deficit hyperactivity disorder in children. Danger is one of overdiagnosis. PMID- 9867378 TI - Attention deficit hyperactivity disorder in children. Multidisciplinary approach to management is needed. PMID- 9867379 TI - Cholesterol lowering diets and coronary heart disease. Dietary advice should focus on promoting antioxidants and the right sort of fats. PMID- 9867380 TI - Cost effectiveness of community leg ulcer clinics. Sensitivity of tools used may explain difference in results between studies. PMID- 9867381 TI - Beef was not "perfectly safe". PMID- 9867382 TI - Food plan needed to reduce deaths from heart disease in Britain. PMID- 9867383 TI - Fever associated with cyclosporin for treating atopic dermatitis. PMID- 9867384 TI - Satisfaction with nurse specialists in breast care clinics. Nurses are not as effective as consultants. PMID- 9867385 TI - Aspirin prophylaxis for vascular disease. Carotid endarterectomy should have been mentioned. PMID- 9867386 TI - Provision of intensive care for children Tertiary centres are unproved. PMID- 9867387 TI - Provision of intensive care for children. Evidence does not support tertiary care. PMID- 9867389 TI - Modern Laser Microscopy in Cellular and Molecular Biology. Proceedings of a workshop. Jena, Germany, 18-21 November 1997. PMID- 9867388 TI - Provision of intensive care for children. Results of Trent and Victoria study are valid. PMID- 9867390 TI - Ultrastructural in situ hybridization: a review of technical aspects. AB - Detection of nucleic acid sequence at the ultrastructural level has allowed us to better understand the expression of genes in some fields of application in cell biology. In situ hybridization at the ultrastructural level can be carried out using three different methods: on vibratome sections before embedding in epoxy resin, on ultrathin frozen section, or on ultrathin section of tissue embedded in hydrophilic resin such as Lowicryl. Before starting the detection of nucleic acid sequences at the electron microscope level, the experimenter has to choose various parameters; the type of tissue fixation, the probe and its label, and the in situ hybridization method, depending on the sensitivity, the resolution and the ultrastructural preservation required. This review of technical aspects, by describing the different methods of ultrastructural in situ hybridization, will help the experimenter to optimize each step of the hybridization procedure. PMID- 9867391 TI - Proceedings of the 3rd International Conference on Advances in Pulmonary Rehabilitation and Management of Chronic Respiratory Failure. Florence, Italy, March 11-14, 1998. PMID- 9867392 TI - Performance. Managed care update. PMID- 9867393 TI - Proceedings of the 8th Symposium on Handling of Environmental and Biological Samples in Chromatography and the 26th Scientific Meeting of the Spanish Group of Chromatography. Almeria, Spain, 26-29 October 1997. PMID- 9867394 TI - Simultaneous determination of fat-soluble vitamins A, D and E and pro-vitamin D2 in animal feeds by one-step extraction and high-performance liquid chromatography analysis. AB - Traditionally, alkaline saponification for the extraction of vitamins and step wise HPLC analyses have been widely used for analysis of fat-soluble vitamins in animal feeds. The objective of present study was to develop an analytical method using one-step extraction and simultaneous determination of the vitamins A, D and E and pro-vitamin D2 in animals feeds b HPLC. Various analytical conditions were tested which included sample particle size extraction solvents, the ratio of solvent to sample, extraction approach, extraction time, N2 protection, prepurification with Sep-Pak cartridge and detection wavelength. The accuracy of the developed method has been proven by comparison with the AOAC method. The reproducibility of the developed method has been substantiated by repeated recovery experiments. The detection limit for the four vitamins was 10 ng/g of feed sample. One of the important properties of the present method is rapidity and ease of use. PMID- 9867395 TI - [Information on the proposals of the Council of the European Economic Community on the protection of workers at risk related to work with dangerous biological objects (on the execution of Article 118(a) of the Treaty of the European Economic Community (EEC) and the refining of Directive 90/679 of the EEC Council)]. PMID- 9867396 TI - Cigarette smoking, asbestos exposure, lung cancer, and sample size. PMID- 9867397 TI - Tracheal deviation as a sign in ill patients: beware ipsilateral pathology. PMID- 9867398 TI - Closed digital artery injury. PMID- 9867399 TI - Waldenstrom macroglobulinaemia presenting as bleeding diathesis with paradoxical coagulation of blood samples. PMID- 9867400 TI - Delayed spinal cord compression in ankylosing spondylitis. PMID- 9867401 TI - Management of sickle cell crisis. British Association for Accident and Emergency Medicine guidelines. PMID- 9867402 TI - Proceedings of the International Symposium on Integrated Ecotoxicology from Molecules/Organisms to Ecosystems. Milano, Italy, June 30-July 2, 1997. PMID- 9867403 TI - Primate evolution--in and out of Africa. PMID- 9867404 TI - Primate evolution--in an out of Africa. PMID- 9867405 TI - Primate evolution--in and out of Africa. PMID- 9867406 TI - Primate evolution--in and out of Africa. PMID- 9867407 TI - Proceedings of the 1st Brain Research Interactive Symposium. Knockouts and mutants: genetically dissecting brain and behavior. San Diego, California, USA. 4 6 November 1998. PMID- 9867408 TI - [Reports presented at the plenary session of the 8th Congress of Czech and Slovak Immunologists in Piestany, 7-10 September 1997]. PMID- 9867409 TI - Report updates recommendations for health workers exposed to HIV. PMID- 9867411 TI - Proceedings of the 6th International Conference on Intelligent Systems for Molecular Biology. Montreal, Quebec, Canada. June 28-July 1, 1998. PMID- 9867410 TI - [High mobility group protein HMG1/HMG2: clinical significance of the autoantibodies]. PMID- 9867412 TI - New infectious disease research funding. PMID- 9867413 TI - Why victims don't talk, why doctors don't ask. 2. PMID- 9867415 TI - Proceedings of the 2nd Oxford International Symposium on the Costs and Benefits of Radiology. San Francisco, California, USA. August 21-23, 1997. PMID- 9867414 TI - Research in radiology residency: a commentary. PMID- 9867416 TI - Divide and conquer: understanding the natural history of cancer. XVth Meeting of the European Association for Cancer Research: five natural histories of human cancer. Stockholm, Sweden. 15-19 August 1998. PMID- 9867417 TI - Life on the bio-pharm: therapeutic proteins from transgenic organisms. Cambridge Healthtech Institute's Transgenic Production of Human Therapeutics. Waltham, MA, USA, 29-30 June 1998. PMID- 9867418 TI - Crystal structure of a new alpha-cyclodextrin hydrate form. Molecular geometry and packing features: disordered solvent contribution. AB - The crystallographic study of a new hydrated form of alpha-cyclodextrin (cyclohexaamylose) is reported. C36H60O30 . 11H2O; space group P2(1)2(1)2(1) with cell constants a = 13.839(3), b = 15.398(3), c = 24.209(7) A; final discrepancy index R = 0.057 for the 5182 observed reflections and 632 refined parameters. Besides four ordered water molecules placed outside alpha-cyclodextrins, the structure shows regions of severely disordered solvent mainly confined in the oligosaccharide cavities. The contribution of the observed disorder has been computed via Fourier inversions of the residual electron density and incorporated into the structure factors in further refinements of the ordered part. The alpha cyclodextrin molecule assumes a relaxed round shape stabilised by a ring sequence of all the six possible O2 ... O3 intramolecular hydrogen bonds. The four ordered water molecules take part in an extensive network of hydrogen bonds (infinite chains and loops) without modifying the scheme of intramolecular H-bonds or the ( )gauche conformation of O-6-H hydroxyl groups. The structure shows a new molecular arrangement, for an "empty" hydrated alpha-cyclodextrin, like that "brick-type" observed for alpha-CD in the iodoanilide trihydrate complex crystallising in an isomorphous cell. PMID- 9867419 TI - ["Hybrid myocardial revascularization": is it a rational therapeutic strategy?]. PMID- 9867420 TI - [Clinical significance of polysomnographic monitoring in cardiology]. PMID- 9867421 TI - WebWatch. Arthritis. PMID- 9867422 TI - LiteratureWatch. PMID- 9867423 TI - Improved estimation of CSA-dipolar coupling cross-correlation rates from laboratory-frame relaxation experiments. AB - We have investigated the underlying assumptions in estimating cross-correlation rates between chemical shift anisotropy (CSA) and dipolar coupling mechanisms in a scalar-coupled two-spin IS system, from laboratory frame relaxation experiments. It has ben shown that for an arbitrary relaxation delay, the difference in relaxation rates of the individual components of an in-phase (or antiphase) doublet is not related to the CSA-dipolar coupling cross-correlation rate in a simple way. This is especially true in the case where the difference in the decay rates of the in-phase and antiphase terms of the density matrix becomes comparable to the magnitude of the scalar coupling between the two spins. Improved means of extracting cross-correlation rates in these cases are presented. PMID- 9867424 TI - Retraction. The TSG101 tumor susceptibility gene is located in chromosome 11 band p15 and is mutated in human breast cancer. PMID- 9867425 TI - Retraction. TAF(II)s mediate activation of transcription in the Drosophila embryo. PMID- 9867426 TI - Body fluid markers for course and activity of disease in multiple sclerosis and 3rd European Charcot Foundation Lecture. Madrid, Spain, 2-4 October 1997. Proceedings. PMID- 9867427 TI - Tap water scald burns in children. 1997. PMID- 9867428 TI - Outcomes after complete immunosuppression withdrawal in liver transplant recipients: rejection-free graft tolerance or late acute rejection? PMID- 9867429 TI - [D.K. Bogorodinskii (the centennial of his birth)]. PMID- 9867430 TI - [Neuroimmune relations in the rat thymus caused by recombinant interleukin-1beta administration]. AB - Interrelations among changes in 11-oxycorticosteroids (11-OSC) in blood plasma, catecholamines (CA), and acetylcholinesterase (AChE) activity in nerve fibres of the thymus, as well as qualitative content and proliferative capacity of thymic and blood cells under the effect of the rat recombinant interleukin-1 beta (IL 1beta) normally and under conditions of hypothalamic-pituitary-adrenocortical depression, were studied. The changes were the most obvious within 15 and 120 min following the IL-2 beta administration: elevation of the 11-OSC, increase in the CA content, and decrease in the AChE activity in thymic nerve fibres. Preliminary administration of dexamethasone prevented the effects. PMID- 9867431 TI - Crystallization of Arthrobacter sp. strain 1C N-(1-D-carboxyethyl)-L- norvaline dehydrogenase and its complex with NAD+. AB - The novel NAD+-linked opine dehydrogenase from a soil isolate Arthrobacter sp. strain 1C belongs to an enzyme superfamily whose members exhibit quite diverse substrate specificities. Crystals of this opine dehydrogenase, obtained in the presence or absence of co-factor and substrates, have been shown to diffract to beyond 1.8 A resolution. X-ray precession photographs have established that the crystals belong to space group P21212, with cell parameters a = 104.9, b = 80.0, c = 45.5 A and a single subunit in the asymmetric unit. The elucidation of the three-dimensional structure of this enzyme will provide a structural framework for this novel class of dehydrogenases to enable a comparison to be made with other enzyme families and also as the basis for mutagenesis experiments directed towards the production of natural and synthetic opine-type compounds containing two chiral centres. PMID- 9867432 TI - A novel approach for the crystallization of soluble proteins using non-ionic surfactants. AB - Crystallization trials using three polyoxyethylene surfactants as precipitating agents are described. Of the eight soluble proteins screened, five were successfully crystallized at the first attempt. These included lysozyme, catalase, ferritin, ribonuclease A and ubiquitin. Further work suggested that these surfactants could also be suitable for cryo-crystallographic analysis of crystals. At the concentrations used in the crystallization trials [10-40%(v/v)], they are capable of promoting the formation of non-crystalline glasses at cryogenic temperatures (77K). This would facilitate crystal mounting and allow the minimization of crystal irradiation damage. Results from this study also suggest that proteins remain stable at high concentrations of these surfactants [40%(w/v)] and over long time periods (>1 month). A number of membrane proteins were also screened for crystallization. These included photosystems I and II and light harvesting complexes I and II from spinach and bacteriorhodopsin from Halobacterium halobium++. The trial s were unsuccessful both in the absence and presence of heptane-1,2,3-triol and over a wide range of surfactant concentrations. PMID- 9867433 TI - Structure of a monoclinic crystal from of cyctochrome b1 (Bacterioferritin) from E. coli. AB - Crystals of E. coli cytochrome b1, alias bacterioferritin, were grown fr om a low ionic strength solution. The resulting monoclniic P21 structure was solved by molecular replacement and refined using noncrystallographi c symmetries applied to the fundamental unit, consisting of two protein subunits and a single haem. From the Patterson self-rotation results it was shown that the asymmetric unit of the monoclinic crystal consists of 12 such dimers and corresponds to a complete, nearly spherical, molecule of bacterioferritin (M4 = 450 kDa) of 432 point-group symmetry. It is thus the most symmetrical cytochrome. As previously determined for the tetragonal form, the haem is located in a special position on a local twofold axis of the dimer. A bimetal centre is also observed within the four helix bundle of each monomer; a metal-binding site is located on the fourfold axis. PMID- 9867434 TI - Carboxypeptidase A: native, zinc-removed and mercury-replaced forms. AB - The crystal structure of the zinc-containing exopeptidase bovine carboxypeptidase A (CPA) has been refined to high resolution, based on a data set collected from a single crystal, incorporating new sequence information based on cloning of the bovine gene. In addition, new refined structures are available for the zinc removed form of the enzyme, apo-CPA, as well as the mercury-replaced form, Hg CPA. The native structure reveals that the zinc-bound water molecule does not appear to more loosely bound than the rest of the zinc ligands, at least when B factor values are considered. Nor is there any evidence for a secondary location of this water molecule. The apo-enzyme structure does not show any density in the place of the removed zinc ion. The only significant change appears to be a chi2 rotation of one zinc histidine ligand to form an ion-pair interaction with a glutamic acid side chain. The structure of Hg-CPA reveals a solvent Tris molecule bound to the mercury cation, as well as an unidentified cation bound to Glu270. The location of this citation agrees with previous proposals for the binding side of inhibitory zinc. These observations may explain some of the differences in kinetics observed in metal- replaced CPA. PMID- 9867435 TI - Structure of an enantiomeric protein, D-monellin at 1.8 A resolution. AB - The D-enantiomer of a potently sweet protein, monellin, has been crystallized and analyzed by X-ray crystallography at 1.8 A resolut ion. Two crystal forms (I and II) appeared under crystallization conditions similar, but not identical, to the crystallization conditions of natural L-monellin. There are four molecules per asymmetric unit in crystal form I and one in crystal form II. Crystal form I is not reproducible and is equivalent to that of monoclinic L-monellin. Intermonomer contacts in crystal form II are very different from those found in natural L monellin crystals. The backbone trace of D-monellin resembles very closely the mirror image of that of L-monellin, but the N- and C-terminus backbones as well as several side-chain conformations of D-monellin are different from those of natural L-monellin. Most of these apparent differences may be attributable to the crystal packing differences. PMID- 9867436 TI - On the ab initio solution of the phase problem or macromolecules at very low resolution. II. Generalized likelihood based approach to cluster discrimination. AB - The multisolution strategies for direct phasing at very low resolution, such as the few atoms model technique, result in a number of alternative phase sets, each of them arising from a cluster of closely related models. Use of a Monte-Carlo type computer procedure is suggested to choose between the possible phase sets. It consists of generating a large number of pseudo-atom models inside the mask defined by a trial phase set and the use of histograms of magnitude correlation to evaluate the masks. It is shown that the procedure may be considered as a generalization of the statistical maximum-likelihood principle and may be used as a powerful supplementary tool in the likelihood-based approaches to the phase problem solution. PMID- 9867437 TI - Neutrophil and macrophage responses to inflammation in the peritoneal cavity of rainbow trout Oncorhynchus mykiss. A light and electron microscopic cytochemical study. AB - The neutrophil and macrophage responses that accompany inflammation were studied in the peritoneal cavity of rainbow trout Oncorhynchus mykiss using light and electron microscopic cytochemistry. Neutrophils of inflammatory peritoneal exudates were alpha-naphthyl butyrate esterase-negative, peroxidase-positive and rich in cytoplasmic glycogen granules. Macrophages were poor in glycogen, esterase-positive and usually peroxidase-negative. Some peroxidase-positive macrophages were due to the transfer to macrophages of neutrophilic peroxidase. The ultrastructural double labelling for glycogen-peroxidase or esterase/peroxidase was most useful for precisely characterising neutrophils and macrophages in the inflamed peritoneal cavities and for correctly labelling peroxidase-positive macrophages. Intraperitoneal injection of casein, Incomplete Freund's Adjuvant (IFA) and live or formol-killed Yersinia ruckeri resulted in a rapid influx of neutrophils, peaking at 24 to 48 h post-injection and reaching values, in the case of live bacteria, 500 x those in the resting, unstimulated peritoneal cavity. Peritoneal macrophages also increased, but the response was slower (peak at 5 d) and with more modest increases in number (7.5 x). Neutrophil and mononuclear cells returned to normal values after 15 d in the case of casein and bacteria, but continued above base values 30 d after the injection of IFA. Conversely, after the injection of phosphate buffered saline, India ink or with sham-injections, very moderate neutrophil and and macrophage responses subsided in a few hours. Phagocytosis of bacteria was studied by light microscopy of preparations stained for peroxidase by a new method which allows for the simultaneous observation of intracellular bacteria and peroxidase staining. When Y. ruckeri was injected into resting peritoneal cavities, bacteria were ingested by the resident macrophages. When the bacteria were injected into cavities with high numbers of neutrophils (due to the previous injection of casein), more neutrophils than macrophages contained bacteria. Results show that the macrophages are the resident phagocytes of the peritoneal cavity of trout, while neutrophils are present in that body cavity in significant numbers only in situations of inflammation and only as long as the inflammation persists. PMID- 9867438 TI - Enterococcus-like infections in Macrobrachium rosenbergii are exacerbated by high pH and temperature but reduced by low salinity. AB - Macrobrachium rosenbergii (10 to 15 g and 8 to 12 g at intermolt) were challenged with an enterococcus-like bacterium (strain KM002) previously isolated and identified as the causal agent of mortality. Challenge doses and conditions of pH, salinity and temperature were varied to determine the influence of environmental factors on the development of disease and mortality. Survival was 100% for the unchallenged control groups in all trials. In pH tests, the onset of mortality was earlier at pH 8.8 to 9.5 than at pH 4.6 to 5.2 and 7.5 to 7.7. Also, as pH 8.8 to 9.5, all challenged prawns died within 6 d in high dose challenge tests. By contrast, 20% of the prawns challenged at pH 4.6 to 5.2 and 7.5 to 7.7 survived. At low dose challenge (5 x 10(4) cfu prawn(-1)), survival increased significantly except at pH 8.8 to 9.5. In salinity tests at 2 challenge doses (1 x 10(6) and 2 x 10(7) cfu prawn(-1)), onset of mortality was earliest at 15 ppt and cumulative mortality was 100% at 15 ppt and 0 ppt. By contrast, survival was 80% at 5 and 10 ppt at the low dose challenge and 40% and 60%, respectively, at the high dose challenge. When the challenge dose was reduced to 5 x 10(4) cfu prawn(-1), survival was not significantly different at different salinity levels. In temperature tests at pH 7.2 to 7.5 and at 2 challenge doses (2 x 10(7) and 4 x 10(7) cfu prawn(-1)), the onset of mortality was earliest at 33 to 34 degrees C and total mortality occurred at 27 to 28 degrees C and 33 to 34 degrees C. By contrast, there were 40% and 20% survivors, respectively, for low and high challenge doses at 30 to 31 degrees C. Reducing the challenge dose to 5 x 10(4) cfu prawn(-1) gave higher survival in all groups. However, survival at 33 to 34 degrees C was still lowest. In similar temperature tests but at pH 8.8 to 9.5, onset of mortality was somewhat accelerated and there was 100% death for all the high challenge groups. At low challenge doses, mortality was lower but still highest in the 33 to 34 degrees C group. Results indicated that mortality of M. rosenbergii caused by this Enterococcus-like bacterium was exacerbated by environmental parameters of temperature and pH different from those known to be optimal for prawn growth. By contrast, low salinity appeared to have a beneficial effect on survival. Further work is needed to determine the mechanisms underlying these effects. PMID- 9867439 TI - Preliminary applicability tests of different methacrylic acid copolymers, type C NF, particularly relevant to spreading and film formation. AB - The intention of this study was to show under which conditions a film forming methacrylic acid copolymer coating excipient, corresponding to the requirements of pharmacopoeia, but obtained from different sources, can be substituted without severe problems. The mechanical properties of the film coats were investigated by dynamic-mechanical thermo-analysis (DMTA) experiments to determine with respect to the glass transition the storage modulus E', the loss modulus E'', and the loss factor tan delta. Further determinations concerned the surface tensions of the different coating dispersions. This attribute plays an important role in spreading, distribution and coalescence of the film forming preparations. Finally by a series of small experimental fluidized bed batches cores containing a model drug were coated with the different methacrylic acid copolymers. The resistance of these coated tablets in 0.1 N HC1 as well as their dissolution rates in artificial intestinal juice were tested. The coatings proved themselves so similar that in this case substitutions of products of different provenance are possible. The determinations of surface tension and the DMTA measurements seem to be useful and reliable preliminary applicability tests. PMID- 9867441 TI - Emerging Infectious Diseases: Meeting the Challenge. Proceedings of a conference. June 1995. PMID- 9867440 TI - A message from the President of IIDSOG-USA. PMID- 9867443 TI - Metabolic modulation of acute myocardial infarction. The ECLA (Estudios Cardiologicos Latinoamerica) Collaborative Group. AB - BACKGROUND: Several trials have been performed in the past using glucose, insulin, and potassium infusion (GIK) for the treatment of acute myocardial infarction (AMI). Because of continuing uncertainty about the potential role of this therapeutic intervention, we conducted a randomized trial to evaluate the impact of a GIK solution during the first hours of AMI. METHODS AND RESULTS: Four hundred seven patients with suspected AMI admitted within 24 hours of symptoms onset were enrolled. In a ratio of 2:1, 268 patients were allocated to receive GIK (high- or low-dose) and 139 to receive control. Phlebitis and serum changes in the plasma concentration of glucose or potassium were observed more often with GIK. A trend toward a nonsignificant reduction in major and minor in-hospital events was observed in patients allocated to GIK. In 252 patients (61.9%) treated with reperfusion strategies, a statistically significant reduction in mortality (relative risk [RR] 0.34; 95% CI: 0.15 to 0.78; 2P=0.008) and a consistent trend toward fewer in-hospital events in the GIK group were observed. CONCLUSIONS: Our results confirm that a metabolic modulation strategy in the first hours of an AMI is feasible, applicable worldwide, and has mild side effects. The statistically significant mortality reduction in patients who underwent a reperfusion strategy might have important implications for the management of AMI patients. It is now essential to perform a large-scale trial to reliably determine the magnitude of benefit. PMID- 9867442 TI - Colorectal follow-up planning modified on the basis of our personal experience. AB - BACKGROUND: The authors, evaluating the disappointing follow-up results in patients suffering from colorectal carcinoma who had undergone surgery for cure, tried a more rational follow-up. METHODS: In a retrospective review about 232 patients who adhered to the follow-up protocol, we evaluated the accuracy rates of CEA, liver ultrasonography and abdominal CT. In the same group of patients. we evaluated the type of correlation between the neoplastic recurrence rate and Astler-Coller's classification. RESULTS: (1) In detecting hepatic metastases CEA levels furnished sensitivity and negative predictive value more than liver ultrasonography (83.3 vs. 77.8% and 98.4 vs. 98%, respectively); (2) in our series, we obtained a lower recurrence rate in classes A+B1 (7.5%) and B2 (20.8%) and higher in C1+C2 (44.4%) and D (66.7%) (p < 0.0 1). CONCLUSION: According to these data we decided to eliminate postoperative liver ultrasonography and customize follow-up protocol on tumor staging and timing of cancer relapse. The authors believe that these changes will not modify the results, but cause less psychophysical stress for the patients and reduce costs by 50%. PMID- 9867444 TI - Magnesium and airway reactivity. PMID- 9867445 TI - Time for a lower blood lead action level for children? PMID- 9867446 TI - Powerful AIDS drugs may promote heart disease. PMID- 9867447 TI - [Success and failure in laparoscopic gastric banding]. PMID- 9867448 TI - [Surgical therapy planning and interventions in AIDS]. PMID- 9867449 TI - Proceedings of the 11th International Symposium on Endoscopic Ultrasonography. Kyoto, Japan, January 10-11, 1998. PMID- 9867450 TI - Of mobile phones and morbidity. PMID- 9867451 TI - Lowering water's octane. PMID- 9867452 TI - New system for seafood safety. PMID- 9867453 TI - One-upping the LD50. PMID- 9867454 TI - Leadership is required for getting American minorities into research. PMID- 9867455 TI - The Environmental Genome Project: suggestions and concerns. PMID- 9867458 TI - Pesticide use in developing countries. PMID- 9867456 TI - Interregional differences undermine sperm trend conclusions. PMID- 9867459 TI - The relentless flu drug. PMID- 9867457 TI - Debate about sperm count decline. PMID- 9867460 TI - EPA's PM2.5 program criticized. PMID- 9867461 TI - Not breathing easier. PMID- 9867463 TI - A reconsideration of stress experiments and population heterogeneity. PMID- 9867462 TI - Upbeat therapy and management of problem tachycardias. Proceedings of a satellite symposium of the XIX Congress of the European Society of Cardiology. Stockholm, Sweden, 24 August 1997. PMID- 9867464 TI - Safety of Biological Products Prepared from Mammalian Cell Culture. Proceedings of a conference. Annecy, France, September 29-October 1, 1996. PMID- 9867465 TI - Simian Virus 40 (SV40): a Possible Human Polyomavirus. Symposium proceedings. Bethesda, Maryland, USA. January 27-28, 1997. PMID- 9867466 TI - ["Materiality of thought" (a hypothesis)]. PMID- 9867467 TI - Development of macromolecular X-ray crystallography and computer aided modelling for genetic engineering and drug design in the Czech and Slovak Republics. COPERNICUS project. PMID- 9867468 TI - Production of amylase by Aspergillus fumigatus utilizing alpha-methyl-D glycoside, a synthetic analogue of maltose, as substrate. AB - A strain of Aspergillus fumigatus isolated from soil was able to produce biomass and high amylase activities in media containing alpha-methyl-D-glucoside (alphaMG), a synthetic analogue of maltose, as the only carbon source. alphaMG was a more effective inducer than starch and maltose at the same concentration: alphaMG cultures produced about 3 times more alpha-amylase and glucoamylase activity than starch cultures. Maximum production of alpha-amylase (60 U/mg) and glucoamylase (130 U/mg) was obtained in 8-10 days alphaMG cultures. PMID- 9867469 TI - Extracellular products as mediators of the formation and detachment of Pseudomonas fluorescens biofilms. AB - Pseudomonas fluorescens B52 produces substantial biofilms at the air/liquid/solid interface of glass coverslips clamped vertically and partly submerged in liquid medium at 21 degrees C. Biofilm formation was maximal ca. 20-50 h after inoculation of the liquid medium and as indicated by environmental scanning electron microscopy (ESEM), contained large numbers of bacterial cells that were embedded within an extensive exopolymeric matrix. Incubation beyond 50 h led to reductions in biofilm which ESEM related primarily to losses of exopolymer. Both biofilm formation and the subsequent decline in exopolymer deposition was more rapid, and occurred to greater extents, when supernatants from two-day old cultures of B52 were used as the initial growth media. The addition of N-acyl hexanoyl homoserine lactone to fresh growth medium had a similar effect upon biofilm formation as using spent culture medium. Homoserine lactones could not be demonstrated in spent culture supernatants by an Agrobacterium tumefaciens bioassay. An exopolysaccharide lyase was detected in spend culture media taken from dense biofilm cultures whose action was specifically directed towards biofilm exopolysaccharide. Results suggest that (i) cell-cell signals such as homoserine lactones are associated with the formation of P. fluorescens biofilms, (ii) the enzymic degradation of exopolymers has a specific role in the detachment of cells under starvation conditions, and (iii) whilst short chain (C6) exogenous homoserines can trigger such response in P. fluorescens, its own signal substance is likely to possess a longer (> C8) fatty acyl chain. PMID- 9867470 TI - Halogenation of aromatic compounds: thermodynamic, mechanistic and ecological aspects. AB - Biological halogenation of aromatic compounds implies the generation of reducing equivalents in the form of e.g. NADH. Thermodynamic calculations show that coupling the halogenation step to a step in which the reducing equivalents are oxidized with a potent oxidant such as O2 or N2O makes the halogenation reaction thermodynamically feasible without the input of additional energy in the form of e.g. NADH. In a current model on the halogenation of tryptophan to 7-chloro-L tryptophan NADH and O2 are proposed as co-substrates in a reaction in which the aromatic compound is oxidized via an epoxide as intermediate. The thermodynamic calculations thus indicate that such a route hinges on mechanistic insights but has no thermodynamic necessity. Furthermore the calculations suggest that halogenation of tryptophan and other aromatic compounds should be possible with N2O, and possibly even with nitrate replacing O2 as the oxidant. PMID- 9867471 TI - Effect of pH on physiology of Metarhizium anisopliae for production of swainsonine. AB - The effect of pH on the production of swainsonine and fungal morphology at different stages of fermentation of Metarhizium anisopliae was investigated. When no control was applied, the pH of the culture dropped from 6.5 to 3.8 within the first 72 hours and the concentration of swainsonine reached 43.3 mg 1(-1). When the pH was held constant either at the beginning or throughout the fermentation, the maximum recorded swainsonine level was only 8.4 mg 1(-1) corresponding with an increase in the formation of pellets. A late pH control applied after 72 hours, resulted in a swainsonine titer of 45.5 mg 1(-1). PMID- 9867472 TI - 8th Mammalian Genetics and Development Workshop. London, United Kingdom, 19-21 November 1997. Abstracts. PMID- 9867473 TI - WAT-free mice: diabetes without obesity. PMID- 9867474 TI - [The cases of plagiarism in "Ginekologia Polska"]. PMID- 9867475 TI - [Publication of a review article in "Ginekologia Polska" identified as a plagiarism from the "British Journal of Obstetrics and Gynecology]. PMID- 9867476 TI - [The publication of "Copper and Zinc concentrations in normal and malignant tissues of ovary, uterus and cervix"]. PMID- 9867477 TI - Short-run tests for determining harmful effects of PCB-containing engine oils on cells. AB - Our main objective was to set up reproducible methods for a rapid determination of harmful effects of PCB-containing engine oils on cells. We used a plate method and Scenedesmus quadricauda, Saccharomyces cerevisiae, Rhodotorula glutinis and Pseudomonas putida as test organisms. PMID- 9867479 TI - Kojic acid production by Aspergillus flavus using gelatinized and hydrolyzed sago starch as carbon sources. AB - Direct conversion of gelatinized sago starch into kojic acid by Aspergillus flavus strain having amylolytic enzymes was carried out at two different scales of submerged batch fermentation in a 250-mL shake flask and in a 50-L stirred tank fermentor. For comparison, fermentations were also carried out using glucose and glucose hydrolyzate from enzymic hydrolysis of sago starch as carbon sources. During kojic acid fermentation of starch, starch was first hydrolyzed to glucose by the action of alpha-amylase and glucoamylase during active growth phase. The glucose remaining during the production phase (non-growing phase) was then converted to kojic acid. Kojic acid production (23.5 g/L) using 100 g/L sago starch in a shake flask was comparable to fermentation of glucose (31.5 g/L) and glucose hydrolyzate (27.9 g/L) but in the 50-L fermentor was greatly reduced due to non-optimal aeration conditions. Kojic acid production using glucose was higher in the 50-L fermentor than in the shake flask. PMID- 9867478 TI - Alteration of cell-wall composition of Fusarium oxysporum by copper stress. AB - A strain of Fusarium oxysporum tolerated copper in the growth medium at concentrations up to 600 mg/L. The optimum growth was obtained at 200 mg Cu/L. The mycelium acquired a blue color in the presence of copper. The copper content of isolated cell walls obtained from mycelium grown in the presence of 600 mg Cu/L was 1.5 times higher than that of cell walls obtained from mycelium grown at 200 mg Cu/L and it contained 2.2 and 3.3% copper at 200 and 600 mg Cu/L, respectively. The amount of protein and total sugars increased in both the mycelium and its isolated cell walls in the presence of copper in the growth medium, chitin was also increased in the cell wall, reaching its maximum amount at 200 mg Cu/L--about 2.4 times higher than without copper. Most of amino acid concentrations in the cell wall were increased in the presence of 200 mg Cu/L and decreased above this concentration. Isoleucine, leucine, tyrosine, phenylalanine, and arginine showed the highest increase at this concentration. The altered cell walls obtained from mycelium grown at 200 and 400 mg Cu/L could rebind individual metals more than the control cell walls could. Rebinding of individual metals was in the order Zn > Fe > Ni > Cu > Co. Rebinding of copper by isolated cell walls depended on pH and temperature. PMID- 9867480 TI - 9th International Workshop on Immunodeficient Animals (IWIDA). Prague, Czech Republic, June 15-18, 1997. Proceedings and abstracts. PMID- 9867481 TI - [Taste function tests and taste disorders]. PMID- 9867482 TI - [Decrease in sperm count of human semen]. PMID- 9867484 TI - Cardiovascular Interventions, Coronary and Peripheral. Transcatheter Cardiovascular Therapeutics 10th annual symposium. Washington, D.C., USA. October 7-11, 1998. Proceedings. PMID- 9867483 TI - Cloning, characterization, and chromosomal localization of human superillin (SVIL). AB - Supervillin is a 205-kDa F-actin binding protein originally isolated from bovine neutrophils. This protein is tightly associated with both actin filaments and plasma membranes, suggesting that it forms a high-affinity link between the actin cytoskeleton and the membrane. Human supervillin cDNAs cloned from normal human kidney and from the cervical carcinoma HeLa S3 predict a bipartite structure with three potential nuclear localization signals in the NH2-terminus and three potential actin-binding sequences in the COOH-terminus. In fact, throughout its length, the COOH-terminal half of supervillin is similar to segments 2-6 plus the COOH-terminal "headpiece" of villin, an actin-binding protein in intestinal microvilli. A comparison of the bovine and human sequences indicates that supervillin is highly conserved at the amino acid level, with 79.2% identity of the NH2-terminus and conservation of three of the four nuclear localization signals found in bovine supervillin. The COOH-terminus is even more conserved, with 95.1% amino acid identity overall and 100% conservation of the villin-like headpiece. Supervillin mRNAs are expressed in all human tissue tested, bu are most abundant in muscle, bone marrow, thyroid gland, and salivary gland; comparatively little message is found in brain. Human supervillin mRNA is approximately 7.5 kb; this message is especially abundant in HeLa S3 cervical carcinoma, SW480 adenocarcinoma, and A549 lung carcinoma cell lines. The human supervillin gene (SVIL) is localized to a single chromosomal locus at 10p11.2, a region that is deleted in some prostate tumors. PMID- 9867485 TI - Interventional vascular therapy: past experiences and future directions. Introduction. PMID- 9867487 TI - Proceedings of the 3rd International Symposium on Monogenea. Brno, Czech Republic, 25-30 August 1997. PMID- 9867486 TI - The role of azithromycin in the treatment of Helicobacter pylori infection - a retrospective report. PMID- 9867488 TI - The Neuropsychology Department at Hawaii State Hospital. PMID- 9867489 TI - EMAP, an echinoderm microtubule-associated protein found in microtubule-ribosome complexes. AB - The major non-tubulin polypeptide found associated with microtubules purified from unfertilized sea urchin eggs by cycles of pH-dependent assembly has a Mr of 77,000. The 77,000 Mr polypeptide is heat- and acid-labile, and is antigenically distinct from the mammalian brain MAPs, MAP-2 and tau. Affinity-purified antiserum against the 77,000 Mr polypeptide was used to survey a variety of cells and tissues for the presence of antigenically related polypeptides. A cross reacting polypeptide, ranging in Mr from 72,000 to 80,000, was found in microtubule preparations from a wide variety of echinoderms, including sea urchins, starfish and sand dollars. Indirect immunofluorescence showed that the polypetide was found in interphase as well as mitotic microtubule arrays. No cross-reacting material was detected in microtubules isolated from marine molluscs, mammalian brain or mouse B16 cultured cells. Because the 77,000 Mr MAP is abundant in echinoderms, we have called it EMAP for echinoderm microtubule associated protein. Although the precise function of the EMAP is not known, our data suggest that the EMAP is involved in the attachment of ribosomes to microtubules. Large numbers of ribosomes are attached to the walls of EMAP containing microtubules, but not EMAP-deficient microtubules. Removal of the EMAP from the microtubule by salt-extraction results in the release of ribosomes from the microtubule, indicating that the EMAP may form part or all of the long tapered stalk that connects these two organelles. PMID- 9867490 TI - Rapid immunochromatographic assay for diagnosis of tuberculosis. PMID- 9867491 TI - Catheter sepsis due to Mycobacterium chelonae. PMID- 9867492 TI - Diagnosis of Q fever. PMID- 9867493 TI - Clinical relevance of serum immunoglobulin A antibodies to Epstein-Barr virus envelope glycoprotein gp350 in nasopharyngeal carcinoma patients. PMID- 9867494 TI - Bartonella henselae-based indirect fluorescence assays are useful for diagnosis of cat scratch disease. PMID- 9867495 TI - Nucleotide sequence at position 1081 of the hemagglutinin-neuraminidasegene in wild-type strains of mumps virus is the most relevant marker of virulence. PMID- 9867496 TI - Symposium in honor of Professor Charles Smith: Breeding Objectives and Strategies. PMID- 9867497 TI - An omission. PMID- 9867499 TI - 7th International Conference on Emergency Medicine. Abstracts. PMID- 9867498 TI - Epithelia cells as regulators of airway inflammation. AB - The past decade has seen major advances in our understanding of the metabolic and biochemical functions of the epithelial cell. As a result, the traditional view of the epithelium as a relatively passive physical barrier to the internal environment has been superseded by the concept that the epithelial plays a key role in regulating airway inflammation. The epithelial cell experts this modulatory effect in several ways. Epithelial cells can function maintain mucosal integrity and to modulate local immune responses. They can also limit inflammatory processes by degrading, or inhibiting, proinflammatory mediators and proteins. However, the epithelium also responds to a range of stimuli by producing biologically active mediators that can influence airway inflammation. These include, but are not limited to a broad range of cytokines and chemokines that can exert profound effects on inflammatory cells, as well as lipid and peptide mediators. This review will highlight some of these aspects of the role of the epithelium in regulating airway inflammation. PMID- 9867500 TI - Florence Nightingale and the enduring legacy of transpersonal human caring. PMID- 9867501 TI - A few great themes. PMID- 9867503 TI - Placental ontogeny of the Australian scincid lizards Niveoscincus coventryi and Pseudemoia spenceri. AB - Models for the evolution of placentation among squamate reptiles have been strongly influenced by early work on one lineage of Australian scincid lizards: one of three lineages that include species with specialized allantoplacentation. The discovery of two types of complex allantoplacentae among species currently assigned to the Eugongylus species group led Weekes ([1935] Proc. Zool. Soc. Lond., 2:625-645) to propose that a graded evolutionary sequence was exemplified by the morphological series of allantoplacental types that existed among Australian skinks. One of the model's strengths is the incorporation of testable inferences of the relationship between placental structure and function. However, descriptions of placental structure for some species are incomplete and subsequent taxonomic revisions have resulted in confusion concerning the species studied. One of the ambiguities is the identity of Lygosoma weekesae (Kinghorn [1929] Proc. Linn. Soc. N.S.W., 54:32-33), one of two species for which the most specialized type of allantoplacenta was defined. The distinguishing characteristic of this placental type is the presence of a structure known as a placentome. Subsequent to the original species description, Lygosoma weekesae was named a synonym for Pseudemoia spenceri (Rawlinson [1974] Mem. Natn. Mus. Vict., 35:87-96), yet the placental description (Weekes [1929] Proc. Linn. Soc. N.S.W., 54:34-60) is thought to be of Niveoscincus coventryi (Rawlinson [1975] Mem. Natn. Mus. Vic., 36:1-16). We describe placental ontogeny of N. coventryi and P. spenceri as a contribution to study of comparative placentation among lygosomatine skinks. We conclude that the placental description for L. weekesae could not have been N. coventryi, because a placentome is not a component of the allantoplacenta of N. coventryi. Further, the allantoplacenta of this species does not conform to previously defined categories for Eugongylus group skinks. The allantoplacenta of P. spenceri contains a placentome that is structurally congruous with the original description of placentation for L. weekesae (Weekes [1929] Proc. Linn. Soc. N.S.W., 54:34-60). Comparison of extraembryonic membrane and placental structure among the four viviparous and one oviparous species of Eugongylus group skinks for which data are available shows that each species exhibits characters that distinguish it from other species, while additional characters are shared among all, or some, species. Based on a recent phylogenetic hypothesis, the distribution of allantoplacental types among these species does not support Weekes' ([1935] Proc. Zool. Soc. Lond., 2:625-645) model of a graded sequence for the evolution of squamate placentation. PMID- 9867502 TI - Comparative biology of sperm storage in female salamanders. AB - Females in seven of the ten families of salamanders possess cloacal glands called spermathecae that store sperm. The annual cycle of sperm storage has been studied by light and electron microscopy in eight species representing five families. In these taxa, we recognized 14 characters associated with the spermathecae and traced their evolution on a phylogeny of salamanders based upon other characters. The plasticity and phyletic significance of the spermathecal characters varied greatly. Plethodontids have complex spermathecae while other families possess simple spermathecae; thus, this character has phyletic value as well as being highly conserved within the Salamandroidea. Other characters, such as carbohydrate histochemistry, are highly plastic and show no obvious phyletic trends. The significance of some of these variable characters, such as duration of sperm storage, is apparent only after including in the analysis other aspects of the reproductive cycle, such as length of the mating season. Additional comparative studies, employing the protocol used in this paper, will help further clarify the relationships between phyletic and functional variability in sperm storage mechanisms in salamanders. PMID- 9867504 TI - Structure, function, and evolution of the oviducts of squamate reptiles, with special reference to viviparity and placentation. AB - In lizards and snakes, the oviducts function in fertilization, sperm storage, egg transport, eggshell deposition, maintenance of the early embryo, and expulsion of the egg or fetus. In viviparous forms they also contribute to placentae responsible for gas exchange and nutrient provision to the fetus. Dissections of species of 30 genera coupled with data from the literature indicate that squamate oviducts vary interspecifically in seven macroscopic features, including the extent and nature of regional differentiation, vascular supply, topographic asymmetry, number of oviducts, vaginal pouches, and relationship to the cloaca. The uterus, infundibulum, and vagina differ histologically in their epithelia, glands, and myometrial layers. Season cyclicity occurs in all three oviductal regions, most prominently in the uterus, and is under endocrinological control. Regional and cytological specializations reflect the diverse functions performed by the oviduct. Definitive evidence for oviductal albumen production and egg resorption is lacking. In viviparous squamates, three uterine specializations may facilitate maternal-fetal gas exchange: an attenuated epithelium, reduced uterine glands (and a reduced shell membrane), and increased vascularization. Contrary to previous reports, pregnant uteri show no epithelial erosion or capillary exposure. Specializations for nutrient provision to the fetus include mucosal hypertrophy, enlarged glandular epithelia, and multicellular glands whose secretions are absorbed by the chorioallantois. Comparisons with other amniotes indicate that squamates inherited the oviduct as an organ with capabilities for egg uptake and transport, fertilization, eggshell deposition, and oviposition. Other features have evolved convergently among squamates: infundibular sperm receptacles, unilateral oviduct loss, uterine gestation, placentation, and specializations for placentotrophy. Cladistic analysis indicates that oviductal features associated with deposition of tertiary egg investments in reptiles reflect evolutionary convergence as well as secondary simplification, rather than a unidirectional trend towards increased specialization. PMID- 9867505 TI - [Molecular biology and molecular genetics in the field of otorhinolaryngology- isolation of genes responsible for deafness]. PMID- 9867506 TI - Evaluating suspected papilledema with ocular echography. PMID- 9867507 TI - [Comment on Letter to the Editor]. PMID- 9867508 TI - [A quarter century ago: a JCC member looks back]. PMID- 9867509 TI - Etherolytic cleavage of 4-(2,4-dichlorophenoxy)butyric acid and 4-(4-chloro-2 methylphenoxy)butyric acid by species of Rhodococcus and Aureobacterium isolated from an alkaline environment. AB - Bacterial strains were isolated from the concrete rubble of a demolished herbicide production plant. The predominant feature of these strains was the etherolytic cleavage of 4-(2,4-dichlorophenoxy)butyric acid (DCPB)1) and 4-(4 chloro-2-methylphenoxy)butyric acid (MCPB) while liberating 2,4-dichlorophenol (DCP) and 4-chloro-2-methylphenol (MCP) respectively. Some of the isolates were identified by 16S rDNA sequence analysis and shown to belong to the genera Aureobacterium sp. (strain K2-17) and Rhodococcus (Rh. erythropolis K2-12). The other strains isolated clustered into these two groups according to fatty acid analysis. Etherolytic cleavage proceeded under neutral to alkaline conditions with an optimum at around pH 8.5. With Aureobacterium sp. No. K2-17, the degradation rate was zero at a pH of 6 but as much as 60% of the maximum activity was observed at pH 10.5. With Rh. erythropolis K2-12, by contrast, pronounced activity was detected at pH 6.5 while degradation was no longer observed at pH 10.5. The maximum rates of cleavage were about 1 mmol DCPB/h.g dry mass with Aureobacterium sp. No. K2-17 and about 0.6 mmol DCPB/h.g dry mass with Rh. erythropolis K2-12. DCPB and MCPB were utilized to the same extent. Substrate cleavage and product formation (DCP) proceeded at almost equal rates with Aureobacterium sp. No. K2-17 and Rh. erythropolis K2-12, which indicates that this compound was not further metabolized. Only phenoxybutyric acid compounds served as substrates; phenoxyacetic acid and phenoxypropionic acid derivatives were not utilized by these strains. PMID- 9867510 TI - Proceedings of the 2nd Annual Conference on Formulations and Drug Delivery. La Jolla, California, USA. October 5-8, 1997. PMID- 9867511 TI - Together for tomorrow: diversity. PMID- 9867512 TI - The growing presence of managed care in rural areas. PMID- 9867513 TI - From the Centers for Disease Control and Prevention. Report on survey regarding collection and use of cause of injury data by states. PMID- 9867514 TI - 11th Annual research meeting of the Kind-Philipp-Foundation for Leukemia Research. Vienna, 11-13 June 1998. Abstracts. PMID- 9867515 TI - MEDNET97. Proceedings of a conference on the internet in medicine. November 1997. PMID- 9867516 TI - [A pediatric cystoscope CI-VS-01-D]. PMID- 9867517 TI - [Device for measuring intraocular pressure through the eyelids]. PMID- 9867518 TI - Association between chloroplast and mitochondrial lineages in oaks. AB - Patterns of chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) variation were studied in 378 populations of oak trees sampled throughout the southern half of France. Six cpDNA haplotypes detected in a previous European survey and three new cpDNA haplotypes were found in this region. Two mitochondrial polymorphisms detected earlier by restriction analysis of PCR-amplified fragments alone, or in combination with single-strand conformation polymorphism (SSCP), were compared with the cpDNA data. Sequencing revealed the nature of the two mitochondrial mutations: a single-base substitution and a 4-bp inversion associated with a 22 bp hairpin secondary structure. The single-base substitution was then analyzed by allele-specific amplification. Results for the two cytoplasmic genomes were combined, which allowed the identification of 12 cpDNA-mtDNA haplotypes. The 4-bp mtDNA inversion has appeared independently in different cpDNA lineages. Given the peculiar nature of this mtDNA mutation, we suggest that intramolecular recombination leading to repeated inversions of the 4-bp sequence (rather than paternal leakage of one of the two genomes) is responsible for this pattern. Furthermore, the geographic locations of the unusual cpDNA-mtDNA associations (due to the inversion) usually do not match the zones of contact between divergent haplotypes. In addition, in southern France, the groupings of populations based on the mtDNA substitution were strictly congruent with those based on cpDNA. Because many populations that are polymorphic for both cpDNA and mtDNA have remained in contact since postglacial recolonization in this area without producing any new combination of cytoplasms involving the mitochondrial substitution, we conclude that paternal leakage is not a significant factor at this timescale. Such results confirm and expand our earlier conclusions based on controlled crosses. PMID- 9867519 TI - The many faces of membership. PMID- 9867520 TI - [Proceedings of the conference on Progress of the Presidential Programme "Russia's Children" and its related federal special programs "The Children of the North" and "The Children from Families of Refugees and Forced Migrants" on the section of prevention of parasitic diseases]. PMID- 9867521 TI - 4th Middle East Symposium on Anaesthesia for Cardiac Patients. Riyadh, Saudi Arabia, 28-29 January 1998. Abstracts. PMID- 9867522 TI - Enterobacter cloacae bloodstream infections associated with contaminated prefilled saline syringes--California, November 1998. PMID- 9867523 TI - Response by the authors of 'Recommendations for spacing of test chemical concentrations in the mouse lymphoma tk mutation essay (MLA)'. PMID- 9867524 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 36-1998. An 11-year-old girl with fever, hypotension, and azotemia. PMID- 9867525 TI - Immunization against Lyme disease. PMID- 9867526 TI - Immunization against Lyme disease. PMID- 9867527 TI - Immunization against Lyme disease. PMID- 9867528 TI - Enoxaparin in neurosurgical patients. PMID- 9867529 TI - Association of mutations in the apolipoprotein B gene with hypercholesterolemia and the risk of ischemic heart disease. PMID- 9867530 TI - Association of mutations in the apolipoprotein B gene with hypercholesterolemia and the risk of ischemic heart disease. PMID- 9867531 TI - Detection of carcinoembryonic antigen messenger RNA in lymph nodes from patients with colorectal cancer. PMID- 9867532 TI - Detection of carcinoembryonic antigen messenger RNA in lymph nodes from patients with colorectal cancer. PMID- 9867533 TI - Detection of carcinoembryonic antigen messenger RNA in lymph nodes from patients with colorectal cancer. PMID- 9867534 TI - Respiratory syncytial virus immune globulin and conflict of interest. PMID- 9867535 TI - Cost as a barrier to medical care in relation to unemployment rates. PMID- 9867536 TI - Iron chelation with oral deferiprone in patients with thalassemia. PMID- 9867537 TI - Iron chelation with oral deferiprone in patients with thalassemia. PMID- 9867538 TI - Iron chelation with oral deferiprone in patients with thalassemia. PMID- 9867539 TI - Iron chelation with oral deferiprone in patients with thalassemia. PMID- 9867540 TI - Iron chelation with oral deferiprone in patients with thalassemia. PMID- 9867541 TI - Iron chelation with oral deferiprone in patients with thalassemia. PMID- 9867542 TI - Iron chelation with oral deferiprone in patients with thalassemia. PMID- 9867543 TI - Diabetes and coronary heart disease. PMID- 9867544 TI - Diabetes and coronary heart disease. PMID- 9867545 TI - Diabetes and coronary heart disease. PMID- 9867546 TI - Diabetes and coronary heart disease. PMID- 9867547 TI - Partial-birth abortion. PMID- 9867548 TI - Partial-birth abortion. PMID- 9867549 TI - Munchausen's microbes? PMID- 9867550 TI - Unruptured intracranial aneurysms--risk of rupture and risks of surgical intervention. AB - BACKGROUND: The management of unruptured intracranial aneurysms requires knowledge of the natural history of these lesions and the risks of repairing them. METHODS: A total of 2621 patients at 53 participating centers in the United States, Canada, and Europe were enrolled in the study, which had retrospective and prospective components. In the retrospective component, we assessed the natural history of unruptured intracranial aneurysms in 1449 patients with 1937 unruptured intracranial aneurysms; 727 of the patients had no history of subarachnoid hemorrhage from a different aneurysm (group 1), and 722 had a history of subarachnoid hemorrhage from a different aneurysm that had been repaired successfully (group 2). In the prospective component, we assessed treatment-related morbidity and mortality in 1172 patients with newly diagnosed unruptured intracranial aneurysms. RESULTS: In group 1, the cumulative rate of rupture of aneurysms that were less than 10 mm in diameter at diagnosis was less than 0.05 percent per year, and in group 2, the rate was approximately 11 times as high (0.5 percent per year). The rupture rate of aneurysms that were 10 mm or more in diameter was less than 1 percent per year in both groups, but in group 1, the rate was 6 percent the first year for giant aneurysms (> or =25 mm in diameter). The size and location of the aneurysm were independent predictors of rupture. The overall rate of surgery-related morbidity and mortality was 17.5 percent in group 1 and 13.6 percent in group 2 at 30 days and was 15.7 percent and 13.1 percent, respectively, at 1 year. Age independently predicted surgical outcome. CONCLUSIONS: The likelihood of rupture of unruptured intracranial aneurysms that were less than 10 mm in diameter was exceedingly low among patients in group 1 and was substantially higher among those in group 2. The risk of morbidity and mortality related to surgery greatly exceeded the 7.5-year risk of rupture among patients in group 1 with unruptured intracranial aneurysms smaller than 10 mm in diameter. PMID- 9867551 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 37-1998. A 41-year-old woman with anemia and severe thrombocytopenia. PMID- 9867552 TI - Human rights and health--the Universal Declaration of Human Rights at 50. PMID- 9867553 TI - Exercise-induced asthma. PMID- 9867554 TI - Exercise-induced asthma. PMID- 9867555 TI - Exercise-induced asthma. PMID- 9867556 TI - Exercise-induced asthma. PMID- 9867557 TI - Lamivudine for chronic hepatitis B. PMID- 9867558 TI - Cure of apparent mineralocorticoid excess by kidney transplantation. PMID- 9867559 TI - Tacrolimus ointment for atopic dermatitis. PMID- 9867560 TI - Tacrolimus ointment for atopic dermatitis. PMID- 9867562 TI - Potential cost savings from legalizing physician-assisted suicide. PMID- 9867561 TI - Potential cost savings from legalizing physician-assisted suicide. PMID- 9867563 TI - Hyperkalemia due to a potassium-based water softener. PMID- 9867564 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 38-1998. A 19-year-old man with the acquired immunodeficiency syndrome and persistent fever. PMID- 9867565 TI - Treatment of acute repetitive seizures. PMID- 9867566 TI - Treatment of acute repetitive seizures. PMID- 9867567 TI - Neurocardiogenic syncope. PMID- 9867568 TI - Case 24-1998: cholesterol atheroembolism. PMID- 9867569 TI - Prevention of preterm birth. PMID- 9867570 TI - Prevention of preterm birth. PMID- 9867571 TI - Prevention of preterm birth. PMID- 9867572 TI - Metformin-induced hemolysis with jaundice. PMID- 9867573 TI - Abciximab and alveolar hemorrhage. PMID- 9867574 TI - Bitter pills to swallow. PMID- 9867575 TI - Bitter pills to swallow. PMID- 9867576 TI - [Bullous amyloidosis in multiple myeloma]. PMID- 9867577 TI - [A gigantic condyloma latum]. PMID- 9867578 TI - [Idiopathic scrotal calcinosis]. PMID- 9867579 TI - [Pemphigoid in a 3-year old child]. PMID- 9867580 TI - [Linear morphea]. PMID- 9867581 TI - [Langerhans-cell histiocytosis]. PMID- 9867582 TI - [Cutaneous angiosarcoma]. PMID- 9867583 TI - Syndrome of acquired aphasia with convulsive disorder in children. 1957. PMID- 9867585 TI - Isolated mucocele in an Onodi cell. AB - A rare case of retrobulbar optic neuropathy caused by an isolated mucocele in an Onodi cell is presented. A 63-year-old man suffering from left recurrent optic neuropathy had shown improved visual acuity with steroid treatment at the age of 48 and 56 years. Impaired visual acuity and blurred vision in the left eye appeared again, and a marked inferior decrease in the left visual field was noted. Although axial computed tomography (CT) showed no abnormal findings, magnetic resonance imaging (MRI) revealed a small squamous-shaped lesion (17 x 17 x 7.3 mm) in the left posterior ethmoidal air cell (Onodi cell) hanging over the left optic nerve. Endoscopic sinus surgery was performed. The optic canal was extruding into the Onodi cell cavity, and the superior bony wall of the canal was eroded by the mucocele. The isolated mucocele in the Onodi cell seemed to be compressing the optic nerve downward an causing retrobulbar optic neuropathy. An Onodi cell lesion, even if it was isolated and small, would cause optic neuropathy. MRI and coronal CT scanning should be performed for the diagnosis and preoperative planning in a case of retrobulbar optic neuropathy. PMID- 9867584 TI - Survival with chronic myelogenous leukemia: the outcome of a patient who had one of the longest survivals. PMID- 9867586 TI - Papers from Symposia of the VII European Multicolloquium of Parasitology. Parma, Italy, 2-6 September 1996. PMID- 9867588 TI - The European Society for Paediatric Infectious Diseases annual awards, 1998. PMID- 9867587 TI - Invited commentary: posterior deformational plagiocephaly. PMID- 9867589 TI - Proceedings of the 1st International Conference on Haemophilus influenzae type b Infection in Asia. Bali, Indonesia, December 17-19, 1996. PMID- 9867590 TI - Long QT syndrome. PMID- 9867591 TI - Screening dipstick urinalysis. PMID- 9867592 TI - Screening dipstick urinalysis. PMID- 9867593 TI - Ventilator care on a general ward. PMID- 9867594 TI - Mother-to-child transmission of hepatitis G virus. PMID- 9867595 TI - Phenytoin dosage. PMID- 9867596 TI - Brandon/Hill selected list of nursing books and journals. PMID- 9867597 TI - Pore performance. PMID- 9867598 TI - Diagnostic imaging. PMID- 9867599 TI - Is it time to learn more about wireless computing? Cutting the ties that bind you to your desk. PMID- 9867600 TI - Carotenohematoporphyrins as tumor-imaging dyes. Synthesis and in vitro photophysical characterization. AB - Multichromophoric dyes for use in tumor imaging have been synthesized and photophysically characterized. Structurally, these dyes are dyads and triads that consist of one or two carotenoid polyenes covalently attached to hematoporphyrin (HP) or hematoporphyrin dimethyl ester (HPDME) moieties via ester linkages. The ground-state absorption of each compound shows that the electronic interaction between the chromophores is small. The fluorescence quantum yield for the dyad monocaroteno-HPDME is 0.033 and the dicaroteno-HPDME triads have yields between 0.016 and 0.007, all of which are reduced with respect to the parent compound HPDME (0.09). Global analysis of the transient fluorescence decays of the dyads and triads requires two exponential components (approximately 5-6 ns and approximately 1-2 ns) to fit the data, while a single exponential component with a lifetime of 9.3 ns describes the decay data of the parent HPDME. Possible mechanisms for the observed porphyrin fluorescence quenching by the nearby carotenoid are discussed. Nanosecond transient absorption reveals a carotene triplet with maximum absorption at 560 nm and a 5.0 microsecond lifetime. No transient was detected at 450 nm, indicating rapid (< or = 10 ns) triplet energy transfer from the hematoporphyrin to the carotenoid moieties in fluid as well as in rigid media. The yield of triplet energy transfer from the porphyrin to the carotenoid moiety is unity. Singlet oxygen (O2(1 delta g), studies support the transient absorption data, as none of these compounds is capable of sensitizing O2(1 delta g). Liposome vesicles were used to study the photophysical characteristics of the dyes in phospholipid membranes. Singlet oxygen was not sensitized by the dyads and triads in liposomes. Transient absorption measurements suggest that the triads are substantially aggregated within the phospholipid bilayer, whereas aggregation in the dyads is less severe. PMID- 9867601 TI - Electron transfer processes in the reactivity of nonsteroidal anti-inflammatory drugs in the ground and excited states. AB - The nonsteroidal anti-inflammatory drugs (NSAID), naproxen, sulindac and indomethacin, were shown to donate electrons to nitro blue tetrazolium (NBT) when irradiated with UV light in deoxygenated aqueous buffer solution (pH 7.4, 30 degrees C). The reaction was monitored spectrophotometrically by the appearance of the diformazan reduction product from NBT. The electron transfer process facilitates the decomposition of the drugs. Naproxen in the presence of NBT is photodegraded principally to the alcohol (2-[1-hydroxyethyl]-6 methoxynaphthalene) at a rate approximately 20-fold faster than when irradiated alone in deoxygenated conditions. The photoproduct from naproxen also participates in the electron transfer to NBT but at a much slower rate than naproxen. Irradiation of sulindac or indomethacin in the presence of NBT caused the slow photoreduction of NBT to diformazan. In the absence of NBT, indomethacin and sulindac are essentially unreactive when irradiated in aqueous solution. The ability of a number of NSAID to act as electron donors in their ground state was studied by observing their oxidation by potassium peroxodisulfate in pH 7.0 phosphate buffer at 50 degrees C. The HPLC analysis of the drug remaining showed that the 2-arylpropionic acid NSAID (naproxen, ibuprofen, ketoprofen and suprofen) reacted at a rate equivalent to the thermal decomposition of peroxodisulfate. The major products were the same as detected in the photooxidation of these drugs, resulting from decarboxylation and oxygen addition but also included a dimeric compound. On the other hand, the NSAID that do not contain the propionic acid substituent all reacted more slowly with peroxodisulfate, enabling specific reaction rate constants to be evaluated. PMID- 9867602 TI - The Electronic Plant Gene Register. PMID- 9867603 TI - [Preservation of E coli strains]. PMID- 9867604 TI - [E coli genomes]. PMID- 9867605 TI - [Measuring apparatus for pH (discussion)]. PMID- 9867606 TI - Rembrandt revealed. PMID- 9867607 TI - [Bronchopulmonary Chronic Obstruction Disease: trends and perspectives. Proceedings of a seminar. France, November 1997]. PMID- 9867608 TI - Authorities for resuscitation in Germany. PMID- 9867609 TI - [Bronchial obstructive diseases. Symposium proceedings. Paris, France, 14 November 1997]. PMID- 9867610 TI - [Corticotherapy and infectious broncho-pulmonary diseases. Workshop proceedings. 13-14 September 1997]. PMID- 9867611 TI - [Use of corticoids in severe extensive lung diseases and adult respiratory distress syndrome (ARDS)]. PMID- 9867612 TI - Everyday speechreading: understanding seen speech action. AB - How does speechreading (lipreading) work at the cognitive and neurobiological level? In this review I summarise work over the last fifteen years that has used everyday speechreading in hearing participants to illuminate issues of cerebral localisation and cognitive function. The implications of this work for deepening our understanding of speechreading in a remedial context are highlighted. PMID- 9867613 TI - [Is the dentist working with one foot in the grave?]. PMID- 9867614 TI - ["Quality legitimizes a free profession". The Interdisciplinary Continuing Education Week "Quality '99" (IFW '99). Interview by Peter Jager]. PMID- 9867615 TI - [Iconography--the monogrammist HF. "The tooth crusher"]. PMID- 9867616 TI - [Titanium, the future is being outlined! 4th International Symposium on Titanium in Dentistry, 2-4 September 1998 at Geneva]. PMID- 9867617 TI - [A reader's letter on "Amalgam, a question of belief"]. PMID- 9867618 TI - [The therapy of benign bone and soft-tissue tumors of the oral area. The annual meeting of the Swiss Association for Oral Surgery and Mouth Diseases (SAKM) of 3 March 1998 in Bern]. PMID- 9867619 TI - [Diagnosis in dental radiology. The annual meeting of the Swiss Society for Dentomaxillofacial Radiology (SGDMFR) of 6 March 1998 in Bern]. PMID- 9867620 TI - [Quality standards in dentistry--a nightmare?]. PMID- 9867621 TI - Leaping leptin. PMID- 9867622 TI - The population slide. PMID- 9867623 TI - Understanding of ears, bristles jumps a notch. PMID- 9867624 TI - Reaction to stem cells: a tale of the ticker. PMID- 9867625 TI - Pasteur recruit resigns in battle over new unit. PMID- 9867626 TI - Genome links typhus bug to mitochondrion. PMID- 9867627 TI - Training viruses to attack cancers. PMID- 9867628 TI - Issues facing the new FDA commissioner. PMID- 9867629 TI - Publishers' honeymoon? PMID- 9867630 TI - Apoptosis and Alzheimer's disease. PMID- 9867631 TI - Genetics of alcoholism. PMID- 9867632 TI - Gathering glycine receptors at synapses. PMID- 9867633 TI - A cellular striptease act. PMID- 9867634 TI - Claim of human-cow embryo greeted with skepticism. PMID- 9867635 TI - AIDS vaccines. India prepares to join U.S., world teams. PMID- 9867636 TI - Can IL-2 smoke out HIV reservoirs? PMID- 9867638 TI - fMRI provides new view of monkey brains. PMID- 9867637 TI - A possible new partner for telomerase. PMID- 9867639 TI - Requiem for life on Mars? Support for microbes fades. PMID- 9867640 TI - Finding life's limits. PMID- 9867641 TI - Training a molecular gun on killer E. coli. PMID- 9867642 TI - New ways to probe the molecules of life. PMID- 9867643 TI - Animal rights: reaching the public. PMID- 9867644 TI - Declining fertility. PMID- 9867645 TI - Declining fertility. PMID- 9867646 TI - Laboratory safety. PMID- 9867647 TI - Patenting genomic technologies. PMID- 9867648 TI - Strengthening the biological weapons convention. PMID- 9867650 TI - DNA replication. Bringing the mountain to Mohammed. PMID- 9867649 TI - Molecular origin of species. PMID- 9867651 TI - Hidden models in biopolymers. PMID- 9867652 TI - Gene therapy. Treating with HIV. PMID- 9867653 TI - Ligand binding. Molecular barbells. PMID- 9867654 TI - No last word on language origins. PMID- 9867655 TI - DNA suggests cultural traits affect whales' evolution. PMID- 9867656 TI - Panel tightens rules on mental disorders. PMID- 9867657 TI - Hairy mice offer hope for baldness remedy. PMID- 9867658 TI - Improving gene transfer into livestock. PMID- 9867660 TI - Cuba's billion-dollar biotech gamble. PMID- 9867659 TI - Outsmarting HIV drug resistance. PMID- 9867661 TI - Clock photoreceptor shared by plants and animals. PMID- 9867662 TI - HIV's early home and inner life. PMID- 9867663 TI - Environmental health: nickel-and-diming it. PMID- 9867664 TI - The when and where of floor plate induction. PMID- 9867665 TI - Quality assurance in radiation oncology. Proceedings of an international symposium. Berlin, May 1998. PMID- 9867666 TI - Acute radiation morbidity and therapeutic consequences. Proceedings of a workshop of the German Society for Radiooncology. Frankfurt, Germany,May 16-17, 1998. PMID- 9867667 TI - [A list of author abstracts of candidate and doctoral dissertations for 1997 deposited in the Central Research Institute of Stomatology and in the State Central Medical Research Library in January-March 1998]. PMID- 9867668 TI - [Occlusal anomalies in the deciduous and mixed bites]. AB - In 311 examines (177 boys and 134 girls) with primary dentition and 535 examinees (285 boys and 250 girls) with mixed dentition in Istria--Croatia, the frequency of orthodontic and occlusive anomalies in regard of space plane and premature extraction of c, m1 and m2 are investigated. The orthodontic anomalies are defected in 46.95%, premature loss in 11.25% and occlusive anomalies in 40.85% of examinees with primary dentition, while in mixed dentition the 58.69% of examinees have orthodontic anomaly, the 17.20% premature loss and the 48.97% of examinees have occlusal anomaly. In both phases of dentition the most frequent are occlusive anomalies in the following planes: sagittal, vertical and sagittal vertical. PMID- 9867669 TI - [New Perspectives on the Management of Lung Cancer with Paclitaxel. Forum report. Barcelona, 14 March]. PMID- 9867670 TI - [Treatment of Advanced Colorectal Cancer: New Acquisitions. Report of a meeting. Rende, 28 June 1998]. PMID- 9867671 TI - [Ibn Al-Jazzar and Tenth Century medicaments]. PMID- 9867672 TI - [Mohammed Ibn Othman As-Siqilly and his work "el Moukhtassar el Farissi"]. PMID- 9867673 TI - [Place of the works of Aziza Othmana in the history of medicine in Tunisia]. PMID- 9867674 TI - [Doctor Bechir Dinguizli. First Tunisian corresponding member of the French Academy of Medicine 1869-1934]. PMID- 9867675 TI - [Medicine in Tunisia from the French Protectorate to the contemporary era]. PMID- 9867676 TI - [Messages of professors Robert Debre and Gilbert Dreyfus addressed in 1952 to their colleagues in Tunisia in the occasion of the Fiftieth anniversary of the Society of Medical Sciences of Tunisia]. PMID- 9867677 TI - [On the Tunisian Medical Society of History of Medicine and Pharmacy. Notes and recollections]. PMID- 9867678 TI - [From Lazaretto to the Rabta Hospital]. PMID- 9867679 TI - [The Prize of the President of the Republic of Tunisia for Inter-Maghreb Medical Societies]. PMID- 9867680 TI - A brief survey of the history of medicine in Tunisia. PMID- 9867681 TI - Isaac Ibn Suleiman al Israili (end IXth-beginning Xth century AD) (end of IIIrd and first half of IVth century of the Hegira). PMID- 9867682 TI - The man and the tools. PMID- 9867683 TI - Population and human reproduction: the Tunisian experience. PMID- 9867684 TI - [In the times of the "Fatimids of Mehdia" "the encyclopedist physician Abu Salt Oumia el Andalussi"]. PMID- 9867685 TI - [The Spanish hospital of Tunisia (Work of the Castilian Order of the Trinitarians) 1720-1818]. PMID- 9867686 TI - [Medicine in Tunisia in the 19th Century]. PMID- 9867687 TI - [Prof. Sedrak Semenovich Sharimanian (on the centenary of his birth)]. PMID- 9867688 TI - [Genrikh Ivanovich Turner (1858-1941)]. PMID- 9867689 TI - [Dissertations defended in 1998]. PMID- 9867690 TI - [The 10th anniversary of the Veterans Organization of Naval Medics]. PMID- 9867691 TI - [A. A. Budko, colonel in the medical service, was appointed on 26 May 1998 to the post of head of the Military Medical Museum of the Ministry of Defense of the Russian Federation]. PMID- 9867692 TI - Possible urticarial reaction to new ultra-filtered Rho immune globulin. PMID- 9867693 TI - [Basic results of studies on the problem of AIDS in 1996]. PMID- 9867694 TI - [A few words concerning the editorial note about the confirmation of plagiarism with regard to eight published articles by Dr. Andrzej Jendryczko]. PMID- 9867695 TI - [The 30th anniversary of the Moscow Branch of the All-Russian Society of Epidemiologists, Microbiologists and Parasitologists]. PMID- 9867696 TI - Belgian consensus guidelines for the management of Helicobacter pylori related upper gastrointestinal diseases. Brussels, 6-7 February 1998. PMID- 9867697 TI - Proceedings of a conference on Mathematical Modeling in Experimental Nutrition. Davis, California, USA. August 17-20, 1997. PMID- 9867698 TI - [Optical theory of vision]. PMID- 9867699 TI - [Interpretation of new test methods and the Journal of Japanese Ophthalmological Society]. PMID- 9867700 TI - [Laws related to organ transplantation and corneal transplantation]. PMID- 9867701 TI - [On the 10th Examination for Specialties by the Japanese Society of Ophthalmology]. PMID- 9867702 TI - In memoriam, Dr. Charles Edwin Butterworth, Jr. PMID- 9867703 TI - The greater challenge. PMID- 9867704 TI - Question of experimental method in article in July AJO/DO. PMID- 9867706 TI - Controlling the epidemic of cardiovascular disease in chronic renal disease: What do we know? What do we need to know? Where do we go from here? Special report from the National Kidney Foundation Task Force on Cardiovascular Disease. PMID- 9867705 TI - John Nutting Farrar 1839-1913. PMID- 9867707 TI - Low-penetrance branches in matrilineal pedigrees with Leber hereditary optic neuropathy. PMID- 9867708 TI - Detecting marker-disease association by testing for Hardy-Weinberg disequilibrium at a marker locus. AB - We review and extend a recent suggestion that fine-scale localization of a disease-susceptibility locus for a complex disease be done on the basis of deviations from Hardy-Weinberg equilibrium among affected individuals. This deviation is driven by linkage disequilibrium between disease and marker loci in the whole population and requires a heterogeneous genetic basis for the disease. A finding of marker-locus Hardy-Weinberg disequilibrium therefore implies disease heterogeneity and marker-disease linkage disequilibrium. Although a lack of departure of Hardy-Weinberg disequilibrium at marker loci implies that disease susceptibilityweighted linkage disequilibria are zero, given disease heterogeneity, it does not follow that the usual measures of linkage disequilibrium are zero. For disease-susceptibility loci with more than two alleles, therefore, care is needed in the drawing of inferences from marker Hardy Weinberg disequilibria. PMID- 9867709 TI - A critique of Borron et al. PMID- 9867710 TI - Human Immunodeficiency Virus Postexposure Management of Healthcare Workers. Symposium proceedings. Atlanta, Georgia, USA. March 4-5, 1996. PMID- 9867711 TI - Time for resolving shoulder dystocia. PMID- 9867712 TI - Increased cesarean delivery rate: no effect on brachial plexus palsy. PMID- 9867713 TI - Obstetric maneuvers for shoulder dystocia. PMID- 9867715 TI - Hyperemesis gravidarum is not associated with hypofunction of the pituitary adrenal axis. PMID- 9867714 TI - Twin-to-twin transfusion syndrome. PMID- 9867716 TI - What is the role of the AV node if the AV delay occurs before it? PMID- 9867717 TI - TNF-alpha-induced selective cerebral endothelial leakage and increased mortality risk in postmyocardial infarction depression. PMID- 9867719 TI - 30 Years with IgE. Proceedings of an international symposium on basic and clinical aspects of atopic allergy. Stockholm, Sweden, 1-2 September 1998. PMID- 9867718 TI - Alcohol-Induced Immunopathology. Proceedings of a symposium. New Orleans, Louisiana, USA. October 17-18, 1997. PMID- 9867720 TI - [Resuscitation in newborn infants]. PMID- 9867721 TI - [Topic on the theme "volume replacement". 6% HES 130/0.4 is the optimum combination]. PMID- 9867722 TI - Assignment of the canine microsatellite ZuBeCa1 to canine chromosome 10q22-q24. PMID- 9867723 TI - [Biology and genetics of iron metabolism abnormalities. 7th Joint Meeting Inserm SFBC. Paris, 27 November 1997]. PMID- 9867724 TI - [Do corticoids have a preventive effect on laryngeal edema after tracheal extubation?]. PMID- 9867725 TI - Use of the acute cardiac ischemia time-insensitive predictive instrument (ACI TIPI) to assist with triage of patients with chest pain or other symptoms suggestive of acute cardiac ischemia. A multicenter, controlled clinical trial. AB - BACKGROUND: Approximately 6 million U.S. patients present to emergency departments annually with symptoms suggesting acute cardiac ischemia. Triage decisions for these patients are important but remain difficult. OBJECTIVE: To test whether computerized prediction of the probability of acute ischemia, used with electrocardiography, improves the accuracy of triage decisions. DESIGN: Controlled clinical trial. SETTING: 10 hospital emergency departments in the midwestern, southeastern, and northeastern United States. PATIENTS: 10689 patients with chest pain or other symptoms suggestive of acute cardiac ischemia. INTERVENTION: The probability of acute ischemia predicted by the acute cardiac ischemia time-insensitive predictive instrument (ACI-TIPI), either automatically printed or not printed on patients' electrocardiograms. MEASUREMENTS: Emergency department triage to a coronary care unit (CCU), telemetry unit, ward, or home. Other measurements were the bed capacity of the CCU relative to that of the telemetry unit; training or supervision status of the triaging physician; and patient diagnoses and outcomes based on clinical, electrocardiographic, and creatine kinase data. RESULTS: For patients without cardiac ischemia, in hospitals with high-capacity CCUs and relatively low-capacity cardiac telemetry units, use of ACI-TIPI was associated with a reduction in CCU admissions from 15% to 12%, a change of -16% (95% CI, -30% to 0%), and an increase in emergency department discharges to home from 49% to 52%, a change of 6% (CI, 0% to 14%; overall P=0.09). Across all hospitals, for patients evaluated by unsupervised residents, use of ACI-TIPI was associated with a reduction in CCU admissions from 14% to 10%, a change of -32% (CI, -55% to 3%); a reduction in telemetry unit admissions from 39% to 31%, a change of -20% (CI, -34% to -2%); and an increase in discharges to home from 45% to 56%, a change of 25% (CI, 8% to 45%; overall P=0.008). Among patients with stable angina, in hospitals with high-capacity CCUs, use of ACI-TIPI was associated with a reduction in CCU admissions from 26% to 13%, a change of -50% (CI, -70% to -17%), and an increase in discharges to home from 20% to 22%, a change of 10% (CI, -29% to 71%; overall P=0.02). At hospitals with high-capacity telemetry units, use of ACI-TIPI was associated with a reduction in telemetry unit admissions from 68% to 59%, a change of -14% (CI, 27% to 1%), and an increase in emergency department discharges to home from 10% to 21%, a change of 100% (CI, 22% to 230%; overall P=0.02). Among patients with acute myocardial infarction or unstable angina, use of ACI-TIPI did not change appropriate admission (96%) to the CCU or telemetry unit at hospitals with high capacity CCUs or telemetry units. CONCLUSIONS: Use of ACI-TIPI was associated with reduced hospitalization among emergency department patients without acute cardiac ischemia. This result varied as expected according to the CCU and cardiac telemetry unit capacities and physician supervision at individual hospitals. Appropriate admission for unstable angina or acute infarction was not affected. If ACI-TIPI is used widely in the United States, its potential incremental impact may be more than 200000 fewer unnecessary hospitalizations and more than 100000 fewer unnecessary CCU admissions. PMID- 9867726 TI - Use of DNA fingerprinting to assess tuberculosis infection control. AB - BACKGROUND: DNA fingerprinting establishes the genetic relatedness of Mycobacterium tuberculosis isolates and has become a powerful tool in tuberculosis epidemiology. OBJECTIVE: To use DNA fingerprinting to assess the efficacy of current tuberculosis infection-control practices. DESIGN: Retrospective molecular and descriptive epidemiologic study. SETTING: A 700-bed urban public hospital that follows the Centers for Disease Control and Prevention (CDC) guidelines for tuberculosis infection control. PATIENTS: 183 patients who had positive cultures for M. tuberculosis from 1 April 1995 to 31 March 1996. RESULTS: 173 of 183 M. tuberculosis isolates from the study period underwent DNA fingerprinting. Fingerprinting revealed that five isolates represented false positive cultures and that 91 (54%) of the remaining 168 isolates were in 15 DNA fingerprinting clusters, which ranged in size from 2 to 29 isolates. Risk factors for clustering were birth in the United States, African-American ethnicity, homelessness, substance abuse, and male sex. Retrospective epidemiologic analysis of inpatient and outpatient visits by the 91 patients who had clustered isolates revealed only one possible instance of patient-to-patient transmission. CONCLUSIONS: The DNA fingerprinting of all M. tuberculosis isolates from a 1-year period revealed one possible instance of nosocomial transmission and five false positive M. tuberculosis cultures. However, these results did not lead to changes in infection-control practices or in clinical care. The study findings do not support the use of DNA fingerprinting for nosocomial tuberculosis surveillance, but they suggest that compliance with the CDC tuberculosis infection-control guidelines may control patient-to-patient transmission in high-risk urban hospitals. PMID- 9867727 TI - Community-acquired bacterial meningitis: risk stratification for adverse clinical outcome and effect of antibiotic timing. AB - BACKGROUND: Community-acquired bacterial meningitis causes substantial morbidity and mortality in adults. OBJECTIVE: To create and test a prognostic model for persons with community-acquired bacterial meningitis and to determine whether antibiotic timing influences clinical outcome. DESIGN: Retrospective cohort study; patients were divided into derivation and validation samples. SETTING: Four hospitals in Connecticut. PATIENTS: 269 persons who, between 1970 and 1995, had community-acquired bacterial meningitis microbiologically proven by a lumbar puncture done within 24 hours of presentation in the emergency department. MEASUREMENTS: Baseline clinical and laboratory features and times of arrival in the emergency department, performance of lumbar puncture, and administration of antibiotics. The target end point was the development of an adverse clinical outcome (death or neurologic deficit at discharge). RESULTS: For the total group, the hospital mortality rate was 27%. Fifty-six of 269 patients (21 %) developed a neurologic deficit, and in 9% the neurologic deficit persisted at discharge. Three baseline clinical features (hypotension, altered mental status, and seizures) were independently associated with adverse clinical outcome and were used to create a prognostic model from the derivation sample. The prediction accuracy of the model was determined by using the concordance index (c-index). For both the derivation sample (c-index, 0.73 [95% CI, 0.65 to 0.81]) and the validation sample (c-index, 0.81 [CI, 0.71 to 0.92]), the model predicted adverse clinical outcome significantly better than chance. For the total group, the model stratified patients into three prognostic stages: low risk for adverse clinical outcome (9%; stage I), intermediate risk (33%; stage II), and high risk (56%; stage III) (P=0.001). Adverse clinical outcome was more common for patients in whom the prognostic stage advanced from low risk (P=0.008) or intermediate risk (P=0.003) at arrival in the emergency department to high risk before administration of antibiotics. CONCLUSIONS: In persons with community-acquired bacterial meningitis, three baseline clinical features of disease severity predicted adverse clinical outcome and stratified patients into three stages of prognostic severity. Delay in therapy after arrival in the emergency department was associated with adverse clinical outcome when the patient's condition advanced to the highest stage of prognostic severity before the initial antibiotic dose was given. PMID- 9867728 TI - Risk for valvular heart disease among users of fenfluramine and dexfenfluramine who underwent echocardiography before use of medication. AB - BACKGROUND: Because uncontrolled echocardiographic surveys suggested that up to 30% to 38% of users of fenfluramine and dexfenfluramine had valvular disease, these drugs were withdrawn from the market. OBJECTIVE: To determine the risk for new or worsening valvular abnormalities among users of fenfluramine or dexfenfluramine who underwent echocardiography before they began to take these medications. DESIGN: Cohort study. SETTING: Academic primary care practices. PATIENTS: 46 patients who used fenfluramine or dexfenfluramine for 14 days or more and had echocardiograms obtained before therapy. MEASUREMENTS: Follow-up echocardiography. The primary outcome was new or worsening valvulopathy, defined as progression of either aortic or mitral regurgitation by at least one degree of severity and disease that met U.S. Food and Drug Administration criteria (at least mild aortic regurgitation or moderate mitral regurgitation). RESULTS: Two patients (4.3% [95% CI, 0.6% to 14.8%]) receiving fenfluramine-phentermine developed valvular heart disease. One had baseline bicuspid aortic valve and mild aortic regurgitation that progressed to moderate regurgitation. The second patient developed new moderate aortic insufficiency. CONCLUSION: Users of diet medications are at risk for valvular heart disease. However, the incidence may be lower than that reported previously. PMID- 9867729 TI - Treatment of refractory Whipple disease with interferon-gamma. PMID- 9867730 TI - Update in neurology. PMID- 9867731 TI - Drug-induced thrombocytopenia: a systematic review of published case reports. AB - PURPOSE: To determine the strength of clinical evidence for individual drugs as a cause of thrombocytopenia. DATA SOURCES: All English-language reports on drug induced thrombocytopenia. STUDY SELECTION: Articles describing thrombocytopenia caused by heparin were excluded from review. Of the 581 articles reviewed, 20 were excluded because they contained no patient case reports. The remaining 561 articles reported on 774 patients. DATA EXTRACTION: Two of the authors used a priori criteria to independently review each patient case report. Two hundred fifty-nine patient case reports were excluded from further review because of lack of evaluable data, platelet count of 100000 cells/microL or more, use of cytotoxic or nontherapeutic agents, occurrence of drug-induced systemic disease, or occurrence of disease in children. For the remaining 515 patient case reports, a level of evidence for the drug as the cause of thrombocytopenia was assigned. Data on bleeding complications and clinical course were recorded. DATA SYNTHESIS: The evidence supported a definite or probable causal role for the drug in 247 patient case reports (48%). Among the 98 drugs described in these reports, quinidine was mentioned in 38 case reports, gold in 11, and trimethoprim sulfamethoxazole in 10. Of the 247 patients described in the case reports, 23 (9%) had major bleeding and 2 (0.8%) died of bleeding. CONCLUSIONS: Many reports of drug-induced thrombocytopenia do not provide evidence supporting a definite or probable causal relation between the disease and the drug. Future patient case reports should incorporate standard criteria to clearly establish the etiologic role of the drug. PMID- 9867732 TI - Care of the medical ethos: reflections on Social Darwinism, racial hygiene, and the Holocaust. PMID- 9867733 TI - From fate to tragedy: the changing meanings of life, death, and AIDS. AB - The advent of highly active antiretroviral therapy (HAART) and quantitative viral load assays has revolutionized the care of HIV-infected patients. However, this paradigm shift has also had unexpected, sometimes adverse consequences that are not always obvious. Before antiretroviral therapy, physicians learned how to accompany patients through their illness; to bear witness to sickness and dying; and to help patients and their families with suffering, closure, and legacy. Since we have become better at treating the virus, a new temptation has emerged to dwell on quantitative aspects of HIV management and monitoring, although the skills that we learned earlier in the epidemic are no less necessary for providing good care. Our new-found therapeutic capabilities should not distract us from the sometimes more difficult and necessary task of simply "being there" for patients for whom HAART is no longer effective. The definition and practice of end-of-life care for patients with AIDS will continue to evolve as AIDS comes to resemble other chronic, treatable, but ultimately fatal illnesses, such as end stage pulmonary disease and metastatic cancer, in which clinicians must continually readdress with their patients the balance of curative and palliative interventions as the disease process unfolds over time. The coming challenge in HIV care will be to encourage the maintenance of a "primary care" mentality-with attention to the larger psychosocial issues, end-of-life care, bereavement, and a focus on the patient as opposed to the illness-alongside our new antiretroviral paradigm. Otherwise, we run the risk of forgetting what we learned about healing, from a disease that we could not cure. PMID- 9867734 TI - Diet-drug debacle. PMID- 9867735 TI - Population screening for hemochromatosis: the evolving role of genetic analysis. PMID- 9867736 TI - Expanding uses of fluoroquinolones: opportunities and challenges. PMID- 9867737 TI - Grapefruit juice and kidney stones. PMID- 9867738 TI - Intensive therapy preserves insulin secretion. PMID- 9867739 TI - Intensive therapy preserves insulin secretion. PMID- 9867740 TI - Pharmacokinetic interaction between saquinavir and cyclosporine. PMID- 9867741 TI - Cutting waste and keeping faith. PMID- 9867742 TI - Women's health textbooks: codifying science and calling for change. PMID- 9867743 TI - Introduction to supplement on Iron Overload, Public Health, and Genetics. PMID- 9867744 TI - Diagnosis of hemochromatosis. AB - If untreated, hemochromatosis can cause serious illness and early death, but the disease is still substantially under-diagnosed. The cornerstone of screening and case detection is the measurement of serum transferrin saturation and the serum ferritin level. Once the diagnosis is suspected, physicians must use serum ferritin levels and hepatic iron stores on liver biopsy specimens to assess patients for the presence of iron overload. Liver biopsy is also used to establish the presence or absence of cirrhosis, which can affect prognosis and management. A DNA-based test for the HFE gene is commercially available, but its place in the diagnosis of hemochromatosis is still being evaluated. Currently, the most useful role for this test is in the detection of hemochromatosis in the family members of patients with a proven case of the disease. It is crucial to diagnose hemochromatosis before hepatic cirrhosis develops because phlebotomy therapy can avert serious chronic disease and can even lead to normal life expectancy. PMID- 9867745 TI - Management of hemochromatosis. Hemochromatosis Management Working Group. AB - The complications of iron overload in hemochromatosis can be avoided by early diagnosis and appropriate management. Therapeutic phlebotomy is used to remove excess iron and maintain low normal body iron stores, and it should be initiated in men with serum ferritin levels of 300 microg/L or more and in women with serum ferritin levels of 200 microg/L or more, regardless of the presence or absence of symptoms. Typically, therapeutic phlebotomy consists of 1) removal of 1 unit (450 to 500 mL) of blood weekly until the serum ferritin level is 10 to 20 microg/L and 2) maintenance of the serum ferritin level at 50 microg/L or less thereafter by periodic removal of blood. Hyperferritinemia attributable to iron overload is resolved by therapeutic phlebotomy. When applied before iron overload becomes severe, this treatment also prevents complications of iron overload, including hepatic cirrhosis, primary liver cancer, diabetes mellitus, hypogonadotrophic hypogonadism, joint disease, and cardiomyopathy. In patients with established iron overload disease, weakness, fatigue, increased hepatic enzyme concentrations, right upper quadrant pain, and hyperpigmentation are often substantially alleviated by therapeutic phlebotomy. Patients with liver disease, joint disease, diabetes mellitus and other endocrinopathic abnormalities, and cardiac abnormalities often require additional, specific management. Dietary management of hemochromatosis includes avoidance of medicinal iron, mineral supplements, excess vitamin C, and uncooked seafoods. This can reduce the rate of iron reaccumulation; reduce retention of nonferrous metals; and help reduce complications of liver disease, diabetes mellitus, and Vibrio infection. This comprehensive approach to the management of hemochromatosis can decrease the frequency and severity of iron overload, improve quality of life, and increase longevity. PMID- 9867746 TI - Prevalence of elevated serum transferrin saturation in adults in the United States. AB - BACKGROUND: A population-based hemochromatosis screening program that uses serum transferrin saturation has been proposed, but few data exist on the number of U.S. adults that such a program would identify for further testing. OBJECTIVE: To determine the prevalence of an initially elevated serum transferrin saturation and the prevalence of concurrently elevated serum transferrin saturation and serum ferritin levels in the adult population of the United States. DESIGN: Nationally representative cross-sectional survey of the noninstitutionalized U.S. civilian population. PARTICIPANTS: 15 839 men and nonpregnant women 20 years of age and older who were examined in the third National Health and Nutrition Examination Survey (1988-1994). MEASUREMENTS: Single measurements of serum transferrin saturations and serum ferritin levels. Cut-off values used to define elevated serum transferrin saturation ranged from greater than 45% to greater than 62%. RESULTS: The prevalence of initially elevated serum transferrin saturation ranged from 1% to 6%. Approximately 11% to 22% of those with elevated serum transferrin saturation had concurrently elevated serum ferritin levels. The prevalence of elevated serum transferrin saturation was lower in women than in men when the same cut-off value was used to define elevated serum transferrin saturation. The prevalence of elevated serum transferrin saturation in non Hispanic black persons and Mexican-Americans was similar to or slightly less than that in non-Hispanic white persons. The prevalence of elevated serum transferrin saturation in persons 20 to 49 years of age was as high as or higher than that in older adults. CONCLUSIONS: A hemochromatosis screening program that uses a cut off value of greater than 60% to define elevated serum transferrin saturation would identify an estimated 1.4 to 2.5 million U.S. adults for further testing. PMID- 9867747 TI - Hemochromatosis-associated mortality in the United States from 1979 to 1992: an analysis of Multiple-Cause Mortality Data. AB - BACKGROUND: Hemochromatosis, which can lead to serious chronic diseases resulting from iron overload, has an estimated prevalence of 50 to 80 cases per 10000 persons. However, little population-based information is available on the impact of hemochromatosis on morbidity and mortality. OBJECTIVE: To evaluate trends over 14 years in deaths and medical conditions associated with hemochromatosis in the United States. DESIGN: We searched Multiple-Cause Mortality Files compiled by the National Center for Health Statistics for the years 1979 to 1992 for all records listing hemochromatosis. We used these data to calculate age-adjusted and age specific mortality rates, identify medical conditions associated with a known diagnosis of hemochromatosis at death, and calculate proportionate mortality ratios for these medical conditions. RESULTS: The listing of hemochromatosis on death certificates increased 60% from 1979 to 1992. Decedents with hemochromatosis were 23, 13, and 5 times more likely to have liver neoplasms, liver disease, and cardiomyopathy, respectively, than were decedents without hemochromatosis. Conversely, decedents with liver neoplasms, liver disease, and cardiomyopathy were 26, 14, and 5 times more likely, respectively, to have hemochromatosis than were decedents without these conditions. Hemochromatosis was 82 times more likely in persons with the combination of liver neoplasms and diabetes and 43 times more likely in those with the combination of liver disease and diabetes than in those without these conditions. CONCLUSIONS: Comparison of the reported prevalence of hemochromatosis among decedents with estimates of prevalence in the general U.S. population suggests that either the penetrance or the recognition of hemochromatosis, or both, is low. Nevertheless, substantial mortality resulting from liver disease, liver neoplasms, cardiomyopathy, and a combination of liver disease and diabetes in patients with hemochromatosis argues for the improved diagnosis and treatment of hemochromatosis in persons with these conditions. PMID- 9867748 TI - Prevalence of hereditary hemochromatosis in 16031 primary care patients. AB - BACKGROUND: Despite evidence from screening studies in northern European populations, the prevalence of hemochromatosis in primary care populations in the United States remains speculative. OBJECTIVE: To establish the feasibility of screening for hemochromatosis and to estimate the prevalence of hemochromatosis in a large primary care population. DESIGN: Cross-sectional prevalence study. SETTING: 22 primary care practices in the Rochester, New York, area. PATIENTS: 16031 ambulatory patients without a previous diagnosis of hemochromatosis. INTERVENTION: Serum transferrin saturation screening tests were offered to all adult patients in participating primary care practices. MEASUREMENTS: Patients with a serum transferrin saturation of 45% or more on initial testing had a serum transferrin saturation test done under fasting conditions and had serum ferritin levels measured. Those who had a fasting serum transferrin saturation of 55% or more and a serum ferritin level of 200 microg/L or more with no other apparent cause were presumed to have hemochromatosis and were offered liver biopsy to confirm the diagnosis. RESULTS: 25 patients had biopsy-proven hemochromatosis; 22 patients met the clinical criteria for hemochromatosis but declined liver biopsy and were classified as having clinically proven hemochromatosis; and 23 patients had a serum transferrin saturation of 55% or more with no identifiable cause, indicating probable hemochromatosis. The prevalence of clinically proven and biopsy-proven hemochromatosis combined was 4.5 per 1000 (95% CI, 3.3 to 5.8 per 1000) in the total sample and 5.4 per 1000 (CI, 4.0 to 7.1 per 1000) in white persons. The prevalence was higher in men than in women (ratio, 1.8:1). CONCLUSIONS: Hemochromatosis is relatively common among white persons. Routine screening of white persons for hemochromatosis should be considered by primary care physicians. PMID- 9867749 TI - Screening for hemochromatosis in primary care settings. AB - Interest in including screening for hemochromatosis in the routine medical care of adults has grown in recent years. In March 1997, at a meeting on iron overload at the Centers for Disease Control and Prevention, the directors of four hemochromatosis screening programs described the major challenges that they faced and the lessons that they learned in implementing their programs. Seven issues were consistently described as important challenges: 1) changes in case definitions of hemochromatosis, 2) selection of screening threshold values and identification of false-positive cases, 3) variability and lack of standardization in screening test measurements, 4) physician education, 5) informed consent and concerns about medical and genetic discrimination, 6) patient compliance with screening and therapy, and 7) incidental detection of iron deficiency. The two programs that have been completed report a prevalence of iron overload from hemochromatosis of 4.2 to 4.5 per 1000 persons screened; this is consistent with findings in the recent literature. All programs report that screening is feasible and propose that hemochromatosis be defined by repeated elevated serum transferrin saturation values(with or without DNA test results) rather than by the clinical outcome of excessive iron in tissue. The goal of screening programs is to diagnose iron status disorders, particularly hemochromatosis, before they lead to iron overload and chronic disease states. Further research is needed on the ability of genetic and phenotypic tests to predict the clinical expression of hemochromatosis. The experiences outlined in this report highlight practical issues that need to be addressed when iron status screening for hemochromatosis is implemented. It is hoped that this information will facilitate similar efforts in other health care settings. PMID- 9867750 TI - Iron overload, public health, and genetics: evaluating the evidence for hemochromatosis screening. AB - Population screening for hemochromatosis done by using the transferrin saturation test has been advocated by experts to permit the initiation of therapeutic phlebotomy before the onset of clinical disease. The discovery of a gene associated with hemochromatosis has made DNA testing another option for screening and diagnosis. In this paper, U.S. Preventive Services Task Force criteria are used to evaluate the evidence for the usefulness of population screening done by using iron measures or genetic testing. Published clinical research offers little evidence to suggest that population screening for hemochromatosis done by using genetic testing improves clinical outcomes. Although one recently discovered mutation, C282Y, accounts for 60% to 92% of cases of the disease in series of patients with hemochromatosis, uncertainties remain about the clinical penetrance of various genotypes; the accuracy of genetic testing; and the ethical, legal, and social effects of genetic testing. Before population screening for hemochromatosis done by using transferrin saturation testing can be recommended, laboratory standardization needs to be addressed and questions about risk for clinical disease in asymptomatic persons with mutations or early biochemical expression of disease require resolution. Evidence from case series suggests that hemochromatosis may be associated with liver cancer, other liver disease, diabetes, bradyarrhythmias, and arthritis. In all studies but one, however, estimation of the magnitude and significance of this risk is limited by lack of adequate comparison groups. The need for population data to answer questions about penetrance among asymptomatic persons should not impede efforts to increase the detection and treatment of hemochromatosis in persons found to have elevated iron measures a family history of hemochromatosis, or consistent early signs and symptoms of the disease. PMID- 9867751 TI - Public health surveillance for hereditary hemochromatosis. AB - The recent realization that hemochromatosis is a common condition has created opportunities to develop unified public health surveillance for this disorder and its complications and to design programs to prevent unnecessary illness and death resulting from this disorder. Public health surveillance for hemochromatosis can be used to measure the magnitude of the problem (for example, to establish the number of persons with evidence of early iron overload); identify research needs; reveal the natural history of the disease; detect changes in health care practices, such as use of screening tests; and evaluate interventions, such as phlebotomy. Existing surveillance has been limited to periodic measurement of morbidity and mortality done by using hospital discharge records, health examination surveys, vital statistics, and data from small research registries. The improvement of surveillance will entail the ongoing collection of information from population-based surveys, such as the Behavioral Risk Factor Surveillance System; the collection of data on provider practices (for example, through the National Ambulatory Medical Care Survey); and the establishment of population based registries. Creating population-based registries requires consensus on case definitions; strategies to encourage case ascertainment and reporting; policies and procedures for protecting privacy and ensuring confidentiality; and partnerships among providers, researchers, and public health officials. Longitudinal data from population-based registries will provide insight into determinants of disease expression, such as pattern or degree of iron overload. This information is critical for developing evidence-based recommendations for population screening, monitoring changes in medical practices, and assessing the effect of preventive measures. PMID- 9867752 TI - Strategies to increase detection of hemochromatosis. AB - As part of the Iron Overload, Public Health and Genetics conference, sponsored by the Centers for Disease Control and Prevention in March 1997, a working group was convened to consider strategies to increase early case detection of hemochromatosis. This group emphasized that the primary public health goal should be to diagnose hemochromatosis before symptoms appear. To reach this goal, education and action need to be targeted to physicians and other health care workers, laboratorians, administrators, payers, and the public. Strategies to disseminate updated information and increase early case detection were prioritized according to expected effectiveness. Strategies targeting physicians are 1) to identify national and local physician-leaders and 2) to educate physicians about hemochromatosis in basic, graduate specialty, and continuing medical education. Strategies aimed at the health system are 1) to encourage laboratories to provide the transferrin saturation test as part of routine laboratory panels and 2) to work with policymakers and payers to allow reimbursement for case detection. Finally, public education is recommended to increase lay support for the early diagnosis of hemochromatosis. Attempts to educate the public should be aimed first at persons who receive diagnoses of hemochromatosis in order to ensure that they are properly treated and then at asymptomatic persons who could be screened as part of health appraisals. Although identifying physician-leaders and educating physicians are the highest priorities, physicians should not be targeted at the exclusion of payers and the public. Simultaneous efforts to reach all groups in appropriate ways should be initiated to provide the interest and infrastructure necessary to decrease morbidity and mortality from hemochromatosis. PMID- 9867753 TI - Research priorities in hereditary hemochromatosis. AB - The Working Group on Research Priorities used a formal nominal group technique to identify and prioritize the specific aims of applied research needed to provide the scientific basis for population screening for iron overload disorders. The most important applied research goal was characterization of the natural history of the relation between genotype and phenotype in hereditary hemochromatosis and other iron overload disorders. Three other important research objectives were development of an optimal approach to screening for iron overload; analyses of the cost-effectiveness of screening; and assessment of the ethical, legal, and social implications of screening. To achieve these specific aims, two research studies were recommended as being of the highest priority: a multicenter, cross sectional, population-based study of the natural history of iron overload and a multicenter, case-control study of patients with disease manifestations potentially attributable to hereditary hemochromatosis in primary care and subspecialty clinics. PMID- 9867754 TI - Sensitivity and specificity of a rapid whole-blood assay for D-dimer in the diagnosis of pulmonary embolism. AB - BACKGROUND: Patients with suspected pulmonary embolism often have nondiagnostic lung scans and may present in circumstances where lung scanning is unavailable. Levels of D-dimer, a fibrin-specific product, are increased in patients with acute thrombosis; this may simplify the diagnosis of pulmonary embolism. OBJECTIVE: To determine the sensitivity and specificity of a whole-blood D-dimer assay in patients with suspected pulmonary embolism and in subgroups of patients with low pretest probability of pulmonary embolism or nondiagnostic lung scans. DESIGN: Prospective cohort. SETTING: Four tertiary care hospitals. PATIENTS: 1177 consecutive patients with suspected pulmonary embolism. MEASUREMENTS: All patients underwent an assessment of pretest probability by use of a standardized clinical model, a D-dimer assay, ventilation-perfusion lung scanning, and bilateral compression ultrasonography. Patients in whom pulmonary embolism was not initially diagnosed were followed for 3 months. Accordingly, patients were categorized as positive or negative for pulmonary embolism. RESULTS: Of the 1177 patients, 197 (17%) were classified as positive for pulmonary embolism. Overall, the D-dimer assay showed a sensitivity of 84.8% and a specificity of 68.4%. In 703 patients (3.4%) with a low pretest probability of pulmonary embolism, the likelihood ratio associated with a negative D-dimer test result was 0.27, resulting in a posterior probability of 1.0% (95% CI, 0.3% to 2.2%). In 698 patients with nondiagnostic lung scans (previous probability, 7.4%), the likelihood ratio associated with a negative D-dimer test result was 0.36, resulting in a posterior probability of 2.8% (CI, 1.4% to 4.8%). CONCLUSIONS: A normal D-dimer test result is useful in excluding pulmonary embolism in patients with a low pretest probability of pulmonary embolism or a nondiagnostic lung scan. PMID- 9867755 TI - Acquisition of Clostridium difficile and Clostridium difficile-associated diarrhea in hospitalized patients receiving tube feeding. AB - BACKGROUND: Clostridium difficile is the most common infectious cause of nosocomial diarrhea, but its role in diarrhea associated with tube feeding has not been rigorously investigated. OBJECTIVE: To determine the incidence of C. difficile acquisition and C. difficile-associated diarrhea in tube-fed and non tube-fed patients. DESIGN: Prospective cohort study. SETTING: A university affiliated Veterans Affairs Medical Center. PATIENTS: 76 consecutive hospitalized, tube-fed patients and 76 hospitalized, non-tube-fed patients. The two cohorts were matched for age, unit location, duration of hospitalization before surveillance, and severity of illness. MEASUREMENTS: Incidence of C. difficile acquisition, incidence of C. difficile-associated diarrhea, and C. difficile restriction endonuclease analysis typing results. RESULTS: More tube fed patients than non-tube-fed patients acquired C. difficile (15 of 76 patients [20%] compared with 6 of 76 patients [8%]; P=0.03) and developed C. difficile associated diarrhea (7 of 76 patients [9%] compared with 1 of 76 patients [1%]; P=0.03). The mean proportion (+/-SD) of surveillance days with diarrhea was greater for tube-fed patients after the development of C. difficile-associated diarrhea than for tube-fed patients without this diarrhea (0.68+/-0.4 compared with 0.22+/-0.2 [95% CI for the mean difference, 0.08 to 0.84]). Postpyloric tube feeding (odds ratio, 3.14 [CI, 1.008 to 9.77]) and duration of surveillance (odds ratio, 1.08 [CI, 1.0009 to 1.16]) were risk factors for the acquisition of C. difficile. Nineteen restriction endonuclease analysis types of C. difficile were identified from 20 patients. CONCLUSIONS: Hospitalized, tube-fed patients, especially those receiving postpyloric tube feeding, are at greater risk for the acquisition of C. difficile and the development of C. difficile-associated diarrhea than are hospitalized, non-tube-fed patients. Clinicians should test for C. difficile in tube-fed patients with diarrhea. PMID- 9867756 TI - Accuracy of death certificates for coding coronary heart disease as the cause of death. AB - BACKGROUND: Death certificates are widely used in epidemiologic and clinical investigations and for national statistics. OBJECTIVE: To examine the accuracy of death certificates for coding coronary heart disease as the underlying cause of death. DESIGN: Community-based inception cohort followed since 1948. SETTING: Framingham, Massachusetts. PATIENTS: 2683 deceased Framingham Heart Study participants. MEASUREMENTS: Sensitivity, specificity, and predictive values of the death certificate. The reference standard was cause of death adjudicated by a panel of three physicians. RESULTS: Among 2683 decedents, the death certificate coded coronary heart disease as the underlying cause of death for 942; the physician panel assigned coronary heart disease for 758. The death certificate had a sensitivity of 83.8% (95% CI, 81.1 % to 86.4%), positive predictive value of 67.4% (CI, 64.4% to 70.4%), specificity of 84.1% (CI, 82.4% to 85.7%), and negative predictive value of 92.9% (CI, 91.7% to 94.1%) for coronary heart disease. The death certificate assigned coronary heart disease in 51.2% of 242 deaths (9.0% of total deaths) for which the physician panel could not determine a cause. Compared with the physician panel, the death certificate attributed 24.3% more deaths to coronary heart disease overall and more than twice as many deaths to coronary heart disease in decedents who were at least 85 years of age. When deaths that were assigned unknown cause by the physician panel were excluded, the death certificate still assigned more deaths to coronary heart disease (7.9% overall and 43.1% in the oldest age group). CONCLUSIONS: Coronary heart disease may be overrepresented as a cause of death on death certificates. National mortality statistics, which are based on death certificate data, may overestimate the frequency of coronary heart disease by 7.9% to 24.3% overall and by as much as two-fold in older persons. PMID- 9867758 TI - Immunoablative high-dose cyclophosphamide without stem-cell rescue for refractory, severe autoimmune disease. AB - BACKGROUND: Immunoablative high-dose cyclophosphamide without stem-cell rescue induces durable, complete remission in most patients with aplastic anemia. OBJECTIVE: To determine the efficacy of high-dose cyclophosphamide in various refractory, severe autoimmune diseases. DESIGN: Prospective phase II study. SETTING: Johns Hopkins University (Baltimore, Maryland) and Hahnemann University (Philadelphia, Pennsylvania). PATIENTS: Eight patients with refractory, severe autoimmune disease. INTERVENTION: Immunoablative high-dose cyclophosphamide (50 mg/kg of body weight per day) for 4 consecutive days. MEASUREMENTS: Clinical and laboratory variables of autoimmune disease. RESULTS: Seven patients improved markedly: Five achieved complete remission and two achieved partial remission. Four patients have remained in continuous complete remission for 3 to 21 months, and two patients in partial remission continue to improve after 14 and 19 months of follow-up. High-dose cyclophosphamide was well tolerated; median times to a neutrophil count of 0.5 x 10(9) cells/L and platelet transfusion independence were 17 and 16 days, respectively. CONCLUSIONS: Immunoablative high-dose cyclophosphamide without stem-cell rescue can induce complete remission in patients with refractory, severe autoimmune disease. Reemergence of marrow function is similar to that seen after autologous transplantation and does not carry the risk for reinfusion of autoaggressive lymphocytes with the autograft. PMID- 9867759 TI - Update in allergy and immunology. AB - Internal medicine has witnessed astounding developments in our understanding of the immune system's role in the pathophysiology of many diseases. The present challenge is to harness this knowledge to develop more effective therapies for immune-related illness. Guidelines for the care of common illnesses such as asthma may encourage appropriate clinical application of advances in our armamentarium against immune disease. PMID- 9867757 TI - Effect of genetic differences in omeprazole metabolism on cure rates for Helicobacter pylori infection and peptic ulcer. AB - BACKGROUND: Omeprazole is metabolized by S-mephenytoin 4'-hydroxylase (CYP2C19) in the liver. In persons with a poor-metabolizer genotype for CYP2C19, the therapeutic efficacy of omeprazole may be increased. OBJECTIVE: To investigate whether CYP2C19 genotype status is associated with cure rates for Helicobacterpylori infection and peptic ulcer achieved by using dual therapy with omeprazole and amoxicillin. DESIGN: Prospective cohort study. SETTING: University hospital and health service center in Hamamatsu, Japan. PATIENTS: 62 patients with peptic ulcer and H. pylori infection. INTERVENTION: Omeprazole and amoxicillin. MEASUREMENTS: CYP2C19 genotype status and cure rates for H. pylori infection and peptic ulcer. RESULTS: Cure rates for H. pylori infection were 28.6% (95% CI, 13.1% to 48.7%), 60% (CI, 38.6% to 83.0%), and 100% (CI, 66.4% to 100%) in the rapid-, intermediate-, and poor-metabolizer groups, respectively. Healing rates for both duodenal and gastric ulcer in the three groups were roughly parallel with cure rates for H. pylori infection. CONCLUSION: The results of the genotyping test for CYP2C19 seem to predict cure of H. pylori infection and peptic ulcer in patients who receive dual therapy with omeprazole and amoxicillin. PMID- 9867760 TI - The role of venous ultrasonography in the diagnosis of suspected deep venous thrombosis and pulmonary embolism. AB - This paper describes the role of venous ultrasonography in the diagnosis of suspected deep venous thrombosis and pulmonary embolism. Inability to compress the common femoral or popliteal vein is usually diagnostic of a first episode of deep venous thrombosis in symptomatic patients (positive predictive value of about 97%). Full compressibility of both of these sites excludes proximal deep venous thrombosis in symptomatic patients (negative predictive value of about 98%). In patients with suspected deep venous thrombosis or in those who present with suspected pulmonary embolism but have a nondiagnostic lung scan, the subsequent risk for symptomatic venous thromboembolism is very low (<2% during 6 months of follow-up) provided that ultrasonography of the proximal veins remains normal in the course of 1 week (suspected deep venous thrombosis) or 2 weeks (suspected pulmonary embolism). Anticoagulation and further diagnostic testing can usually be safely withheld in these situations. Venous ultrasonography is much less reliable for the diagnosis of asymptomatic, isolated distal, and recurrent deep venous thrombosis than for the diagnosis of a first episode of proximal deep venous thrombosis in symptomatic patients. Clinical evaluation of the probability of deep venous thrombosis or pulmonary embolism, preferably by using a validated clinical model, complements venous ultrasonographic findings and helps to identify patients who would benefit from additional (often invasive) diagnostic testing. Thus, venous ultrasonography is thought to be a very valuable test for the diagnosis and management of patients with suspected deep venous thrombosis or pulmonary embolism. PMID- 9867761 TI - Vulnerable plaque: pathobiology and clinical implications. AB - PURPOSE: To review the pathobiology and clinical implications of vulnerable coronary atherosclerotic plaques and to discuss the identification of vulnerable plaques and mechanisms of plaque stabilization. DATA SOURCES: English-language articles in the MEDLINE database that were published from 1966 to the present, identified by using the terms atherosclerotic plaque, myocardial revascularization, and plaque stabilization. Selected references cited in identified articles were reviewed. STUDY SELECTION: Experimental, clinical, and basic research studies related to coronary atherosclerotic plaques. DATA SYNTHESIS: Rupture at the site of a vulnerable atherosclerotic plaque is the most frequent cause of acute coronary syndromes. Typically, such plaque does not cause high-grade stenosis and has a large lipid core and a thin fibrous cap that is often infiltrated by inflammatory cells. Mechanical stresses contribute to plaque vulnerability, and certain triggers may cause plaque disruption directly. The most important consequence of plaque rupture is thrombosis. No method reliably identifies plaques prone to rupture. The reduction of coronary events by lipid lowering agents despite only modest luminal changes suggests that these agents have a plaque-stabilizing effect. Surgical or percutaneous revascularization does not address the basic biology of coronary atherosclerosis and therefore may have little effect on plaque vulnerability. CONCLUSIONS: Improved understanding of the biology of atheromatous plaques has led to the concept of plaque vulnerability. Identification and stabilization of vulnerable plaques are important new directions in the treatment of coronary atherosclerosis. The relative benefits of aggressive medical therapy aimed at plaque stabilization should be compared with those of revascularization in the management of chronic coronary artery disease. PMID- 9867762 TI - The persuasive appeal of alternative medicine. AB - Alternative medicine has a major presence and persuasive attraction in the industrialized western world. The extent to which these practices have clinical efficacy according to biomedical criteria is a matter of ongoing research and debate. It may be that independent of any such efficacy, the attraction of alternative medicine is related to the power of its underlying shared beliefs and cultural assumptions. The fundamental premises are an advocacy of nature, vitalism, "science," and spirituality. These themes offer patients a participatory experience of empowerment, authenticity, and enlarged self-identity when illness threatens their sense of intactness and connection to the world. A discussion of these themes may enable conventionally trained clinicians to better understand their patients' attraction to and acceptance of alternative medical therapies. PMID- 9867763 TI - Fifty years of death certificates: the Framingham Heart Study. PMID- 9867764 TI - Weighing the alternatives: lessons from the paradoxes of alternative medicine. PMID- 9867765 TI - For love of karaoke. PMID- 9867766 TI - On being a patient. In between. PMID- 9867767 TI - HIV testing in pregnant women. PMID- 9867768 TI - Protein supplements after osteoporotic hip fracture. PMID- 9867769 TI - Protein supplements after osteoporotic hip fracture. PMID- 9867770 TI - Electron-beam computed tomography for evaluating coronary artery disease. PMID- 9867771 TI - Electron-beam computed tomography for evaluating coronary artery disease. PMID- 9867772 TI - Predictors of systemic embolism in mitral stenosis. PMID- 9867773 TI - Sinusitis and gastroesophageal reflux disease. PMID- 9867774 TI - Intravenous immune globulin in autoimmune uveitis. PMID- 9867775 TI - In vivo local thermography of coronary artery atherosclerotic plaques in humans. PMID- 9867776 TI - Fatal hepatotoxicity associated with troglitazone. PMID- 9867777 TI - Terminal dehydration as an alternative to physician-assisted suicide. PMID- 9867778 TI - Terminal dehydration as an alternative to physician-assisted suicide. PMID- 9867779 TI - Terminal dehydration as an alternative to physician-assisted suicide. PMID- 9867780 TI - Terminal dehydration as an alternative to physician-assisted suicide. PMID- 9867781 TI - The genie in the bottle. PMID- 9867782 TI - Accuracy of substituted judgments in patients with terminal diagnoses. PMID- 9867783 TI - Gram stain. PMID- 9867784 TI - Solid organ transplantation. 1: Many problems, new solutions. PMID- 9867785 TI - New approaches for treatment of systemic lupus erythematosus. AB - Each of the new approaches to treating SLE offers some hope in the ongoing effort to respond effectively to the challenges of this debilitating and often lethal condition. Research is extremely active and promises to accelerate as more is learned. PMID- 9867787 TI - [Acute and chronic sphenoidal sinusitis]. PMID- 9867786 TI - Use of a clinical model for safe management of patients with suspected pulmonary embolism. AB - BACKGROUND: The low specificity of ventilation-perfusion lung scanning complicates the management of patients with suspected pulmonary embolism. OBJECTIVE: To determine the safety of a clinical model for patients with suspected pulmonary embolism. DESIGN: Prospective cohort study. SETTING: Five tertiary care hospitals. PATIENTS: 1239 inpatients and outpatients with suspected pulmonary embolism. INTERVENTIONS: A clinical model categorized pretest probability of pulmonary embolism as low, moderate, or high, and ventilation perfusion scanning and bilateral deep venous ultrasonography were done. Testing by serial ultrasonography, venography, or angiography depended on pretest probability and lung scans. MEASUREMENTS: Patients were considered positive for pulmonary embolism if they had an abnormal pulmonary angiogram, abnormal ultrasonogram or venogram, high-probability ventilation-perfusion scan plus moderate or high pretest probability, or venous thromboembolic event during the 3 month follow-up. All other patients were considered negative for pulmonary embolism. Rates of pulmonary embolism during follow-up in patients who had a normal lung scan and those with a non-high-probability scan and normal serial ultrasonogram were compared. RESULTS: Pretest probability was low in 734 patients (3.4% with pulmonary embolism), moderate in 403 (27.8% with pulmonary embolism), and high in 102 (78.4% with pulmonary embolism). Three of the 665 patients (0.5% [95% CI, 0.1% to 1.3%]) with low or moderate pretest probability and a non-high probability scan who were considered negative for pulmonary embolism had pulmonary embolism or deep venous thrombosis during 90-day follow-up; this rate did not differ from that in patients with a normal scan (0.6% [CI, 0.1% to 1.8%]; P > 0.2). CONCLUSION: Management of patients with suspected pulmonary embolism on the basis of pretest probability and results of ventilation-perfusion scanning is safe. PMID- 9867788 TI - [Case for diagnosis. Ectopic cutaneous schistosomiasis]. PMID- 9867789 TI - [100th French Congress of Surgery. Paris, 5-7 October 1998. Abstracts]. PMID- 9867791 TI - [28th Annual meeting of the Scoliosis Society of Quebec. Sainte-Foy, Quebec, Canada. 24-25 April 1998. Abstracts]. PMID- 9867790 TI - [Montreal Surgical Conference. 27 November 1998. Montreal, Canada. Abstracts]. PMID- 9867792 TI - Historical yearly usage of glycopeptides for animals and humans: the American European paradox revisited. PMID- 9867793 TI - Revised sequence of OtrB (tet347) tetracycline efflux protein from Streptomyces rimosus. PMID- 9867794 TI - Novel combination of mutations in the DNA gyrase and topoisomerase IV genes in laboratory-grown fluoroquinolone-resistant Shigella flexneri mutants. PMID- 9867795 TI - History corner. Pros and cons of counting sponges. PMID- 9867796 TI - XIIth International Conference on the Physiology of Food and Fluid Intake (IUOS Satellite) with the 1998 meeting of the Society for the Study of Ingestive Behavior. Pecs, Hungary, 5-8 July 1998. Abstracts. PMID- 9867798 TI - Annual scientific meeting of the Australian Society for Geriatric Medicine. Melbourne, Australia, 11-12 May 1998. Abstracts. PMID- 9867797 TI - Proceedings of Conference on Cardiac and Vascular Surgery: Neurobehavioral Assessment, Physiological Monitoring and Cerebral Protective Strategies. Outcomes '98. Key West, Florida, USA. May 27-31, 1998. PMID- 9867799 TI - Coronary reperfusion. Proceedings of a symposium. Sydney, Australia, July 1996. PMID- 9867801 TI - Prions of mammals and fungi minireview series. PMID- 9867800 TI - Glucose induces expression of stearoyl-CoA desaturase in 3T3-L1 adipocytes. AB - Stearoyl-CoA desaturase (SCD; EC 1.14.99.5) is a key enzyme in the synthesis polyunsaturated fatty acids. Liver and ose tissue are the predominant sites of SCD expression. Regulation of tic SCD by various nutritional and hormonal ors, such as insulin, dietary carbohydrates and polyunsaturated fatty s, has been well documented. Little is known, ver, about adipocyte SCD regulation despite high levels of SCD activity adipose tissue. The present study was gned to investigate SCD regulation in adipocytes by examining the cts of glucose and insulin on SCD expression. We rt here that glucose availability directly increased SCD gene scription in 3T3-L1 adipocytes. This response was pendent of insulin, and insulin alone in the absence of glucose had no ct on SCD mRNA levels. SCD thus represents a l model in which to investigate the mechanisms of direct regulation of expression by glucose in adipose cells. PMID- 9867802 TI - Molecular genetics of transmissible spongiform encephalopathies. PMID- 9867803 TI - Nix and Nip3 form a subfamily of pro-apoptotic mitochondrial proteins. AB - We have identified Nix, a homolog of the E1B 19K/Bcl-2 binding and pro-apoptotic protein Nip3. Human and murine Nix have a 56 and 53% amino acid identity to human and murine Nip3, respectively. The carboxyl terminus of Nix, including a transmembrane domain, is highly homologous to Nip3 but it bears a longer and distinct asparagine/proline-rich N terminus. Human Nip3 maps to chromosome 14q11.2-q12, whereas Nix/BNip3L was found on 8q21. Nix encodes a 23. 8-kDa protein but it is expressed as a 48-kDa protein, suggesting that it homodimerizes similarly to Nip3. Following transfection, Nix protein undergoes progressive proteolysis to an 11-kDa C-terminal fragment, which is blocked by the proteasome inhibitor lactacystin. Nix colocalizes with the mitochondrial matrix protein HSP60, and removal of the putative transmembrane domain (TM) results in general cytoplasmic and nuclear expression. When transiently expressed, Nix and Nip3 but not TM deletion mutants rapidly activate apoptosis. Nix can overcome the suppressers Bcl-2 and Bcl-XL, although high levels of Bcl-XL expression will inhibit apoptosis. We propose that Nix and Nip3 form a new subfamily of pro apoptotic mitochondrial proteins. PMID- 9867804 TI - Interaction of the integrin beta1 cytoplasmic domain with ICAP-1 protein. AB - In a yeast two-hybrid screen, a protein named ICAP-1 (beta1 integrin cytoplasmic domain associated protein) associated with the integrin beta1 cytoplasmic tail but not with tails from three other integrin beta subunits (beta2, beta3, and beta5) or from seven different alpha subunits. Likewise in human cells, ICAP-1 associated specifically with the beta1 but not beta2, beta3, or beta5 tails. The carboxyl-terminal 14 amino acids of beta1 were critical for ICAP-1 interaction. ICAP-1 is a ubiquitously expressed protein of 27 and 31 kDa, with the smaller form being preferentially solubilized by Triton X-100. Phosphorylation of both 27 and 31-kDa forms was constitutive but was increased by 1.5-2-fold upon cell spreading on fibronectin, compared with poly-L-lysine. Also, ICAP-1 contributes to beta1 integrin-dependent migration because (i) ICAP-1 transfection markedly increased chemotactic migration of COS7 cells through fibronectin-coated but not vitronectin-coated porous filters, and (ii) support of beta1-dependent cell migration (in Chinese hamster ovary cells transfected with various wild type and mutant beta1 forms) correlated with ICAP-1 association. In summary, ICAP-1 (i) associates specifically with beta1 integrins, (ii) is phosphorylated upon beta1 integrin-mediated adhesion, and (iii) may regulate beta1-dependent cell migration. PMID- 9867805 TI - Sp3 and Sp4 can repress transcription by competing with Sp1 for the core cis elements on the human ADH5/FDH minimal promoter. AB - The human alcohol dehydrogenase 5 gene (also known as the formaldehyde dehydrogenase gene, ADH5/FDH) has a GC-rich promoter with many sites at which transcription factors bind. A minimal promoter extending from -34 base pairs (bp) to +61 bp directs high levels of transcription in several different cells, consistent with the ubiquitous expression of the gene. Nearly the entire minimal promoter can be bound by Sp1. We analyzed the transcriptional regulation of ADH5/FDH by members of the Sp1 multigene family. Two core cis-elements (-22 bp to +22 bp) had the highest affinity for Sp1. Mutagenesis revealed that these cis elements are critical for transcriptional activation. The zinc-finger domains of Sp3 and Sp4 also bind selectively to the core cis-elements. In Drosophila SL2 cells, which lack endogenous Sp1, the minimal promoter cannot drive transcription. Introduction of Sp1 activated transcription over 50-fold, suggesting that Sp1 is critical in the initiation of transcription. Neither Sp3 nor Sp4 was able to activate transcription in those cells, and transcriptional activation by Sp1 was repressed by Sp3 or Sp4. These data suggest that Sp3 and Sp4 can repress transcription by competing with Sp1 for binding to the core cis elements. The content of Sp1, Sp3, and Sp4 in different cells may be critical factors regulating transcription of the ADH5/FDH gene. PMID- 9867806 TI - Interaction of the cyclic antimicrobial cationic peptide bactenecin with the outer and cytoplasmic membrane. AB - Bactenecin, a 12-amino acid cationic antimicrobial peptide from bovine neutrophils, has two cysteine residues, which form one disulfide bond, making it a cyclic molecule. To study the importance of the disulfide bond, a linear derivative Bac2S was made and the reduced form (linear bactenecin) was also included in this study. Circular dichroism spectroscopy showed that bactenecin existed as a type I beta-turn structure regardless of its environment, while the reduced form and linear bactenecin adopted different conformations according to the lipophilicity of the environment. Bactenecin was more active against the Gram negative wild type bacteria Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhimurium than its linear derivative and reduced form, while all three peptides were equally active against the outer membrane barrier-defective mutants of the first two bacteria. Only the two linear peptides showed activity against the Gram-positive bacteria Staphylococcus epidermidis and Enterococcus facaelis. Bactenecin interacted well with the outer membrane and its higher affinity for E. coli UB1005 lipopolysaccharide and improved ability to permeabilize the outer membrane seemed to account for its better antimicrobial activity against Gram-negative bacteria. The interaction of bactenecin with the cytoplasmic membrane was determined by its ability to dissipate the membrane potential by using the fluorescence probe 3, 3-dipropylthiacarbocyanine and an outer membrane barrier-defective mutant E. coli DC2. It was shown that the linear derivative and reduced form were able to dissipate the membrane potential at much lower concentrations than bactenecin despite the similar minimal inhibitory concentrations of all three against this barrier-defective mutant. PMID- 9867807 TI - ATP synthase of yeast mitochondria. Isolation of subunit j and disruption of the ATP18 gene. AB - The subunit composition of the mitochondrial ATP synthase from Saccharomyces cerevisiae was analyzed using blue native gel electrophoresis and high resolution SDS-polyacrylamide gel electrophoresis. We report here the identification of a novel subunit of molecular mass of 6,687 Da, termed subunit j (Su j). An open reading frame of 127 base pairs (ATP18), which encodes for Su j, was identified on chromosome XIII. Su j does not display sequence similarity to ATP synthase subunits from other organisms. Data base searches, however, identified a potential homolog from Schizosaccharomyces pombe with 51% identity to Su j of S. cerevisiae. Su j, a small protein of 59 amino acid residues, has the characteristics of an integral inner membrane protein with a single transmembrane segment. Deletion of the ATP18 gene encoding Su j led to a strain (Deltasu j) completely deficient in oligomycin-sensitive ATPase activity and unable to grow on nonfermentable carbon sources. The presence of Su j is required for the stable expression of subunits 6 and f of the F0 membrane sector. In the absence of Su j, spontaneously arising rho- cells were observed that lacked also ubiquinol cytochrome c reductase and cytochrome c oxidase activities. We conclude that Su j is a novel and essential subunit of yeast ATP synthase. PMID- 9867808 TI - Comparison of class B scavenger receptors, CD36 and scavenger receptor BI (SR BI), shows that both receptors mediate high density lipoprotein-cholesteryl ester selective uptake but SR-BI exhibits a unique enhancement of cholesteryl ester uptake. AB - Scavenger receptor BI (SR-BI) mediates the selective uptake of high density lipoprotein (HDL) cholesteryl ester (CE), a process by which HDL CE is taken into the cell without internalization and degradation of the HDL particle. The biochemical mechanism by which SR-BI mediates the selective uptake of HDL CE is poorly understood. Given that CE transfer will occur to some extent from HDL to protein-free synthetic membranes, one hypothesis is that the role of SR-BI is primarily to tether HDL close to the cell surface to facilitate CE transfer from the particle to the plasma membrane. In the present study, this hypothesis was tested by comparing the selective uptake of HDL CE mediated by mouse SR-BI (mSR BI) with that mediated by rat CD36 (rCD36), a closely related class B scavenger receptor. Both mSR-BI and rCD36 bind HDL with high affinity, and both receptors mediate HDL CE selective uptake. However, SR-BI mediates selective uptake of HDL CE with a 7-fold greater efficiency than rCD36. HDL CE selective uptake mediated by rCD36 is dependent on HDL binding to the receptor, since a mutation that blocks HDL binding also blocks HDL CE selective uptake. These data lead us to hypothesize that one component of HDL CE selective uptake is the tethering of HDL particles to the cell surface. To explore the molecular domains responsible for the greater efficiency of selective uptake by mSR-BI, we compared binding and selective uptake among mSR-BI, scavenger receptor BII, and various chimeric receptors formed from mSR-BI and rCD36. The results show that the extracellular domain of mSR-BI is essential for efficient HDL CE uptake, but the C-terminal cytoplasmic tail also has a major influence on the selective uptake process. PMID- 9867809 TI - Effect of phosphorylation on activities of Rap1A to interact with Raf-1 and to suppress Ras-dependent Raf-1 activation. AB - Rap1A is phosphorylated by cAMP-dependent protein kinase (PKA), and this phosphorylation has been shown to modulate its interaction with other proteins. However, it is not known whether Rap1A phosphorylation is involved in regulation of its cellular functions, including suppression of Ras-dependent Raf-1 activation. We have previously shown that this suppressive activity of Rap1A is attributable to its greatly enhanced ability to bind to the cysteine-rich region (CRR, residues 152-184) of Raf-1 compared with that of Ras. Here, we show that phosphorylation of Rap1A by PKA abolished its binding activity to CRR. Furthermore, a mutant Rap1A(S180E), whose sole PKA phosphorylation residue, Ser 180, was substituted by an acidic residue, Glu, to mimic its phosphorylated form, failed to suppress Ras-dependent Raf-1 activation in COS-7 cells. These results indicate that the CRR binding activity and the Ras-suppressive function of Rap1A can be modulated through phosphorylation and suggest that Rap1A may function as a PKA-dependent regulator of Raf-1 activation, not merely as a suppressor. PMID- 9867810 TI - Compensatory changes in GroEL/Gp31 affinity as a mechanism for allele-specific genetic interaction. AB - Previous work has shown that the GroEL-GroES interaction is primarily mediated by the GroES mobile loop. In bacteriophage T4 infection, GroES is substituted by the gene 31-encoded cochaperonin, Gp31. Using a genetic selection scheme, we have identified a new set of mutations in gene 31 that affect interaction with GroEL; all mutations result in changes in the mobile loop of Gp31. Biochemical analyses reveal that the mobile loop mutations alter the affinity between Gp31 and GroEL, most likely by modulating the stability of the GroEL-bound hairpin conformation of the mobile loop. Surprisingly, mutations in groEL that display allele-specific interactions with mutations in gene 31 alter residues in the GroEL intermediate domain, distantly located from the mobile loop binding site. The observed patterns of genetic and biochemical interaction between GroES or Gp31 and GroEL point to a mechanism of genetic allele specificity based on compensatory changes in affinity of the protein-protein interaction. Mutations studied in this work indirectly alter affinity by modulating a folding transition in the Gp31 mobile loop or by modulating a hinged conformational change in GroEL. PMID- 9867811 TI - Calcium-dependent oligomerization of synaptotagmins I and II. Synaptotagmins I and II are localized on the same synaptic vesicle and heterodimerize in the presence of calcium. AB - Synaptotagmins constitute a large family of membrane proteins characterized by their distinct distributions and different biochemical features. Genetic evidence suggests that members of this protein family are likely to function as calcium sensors in calcium-regulated events in neurons, although the precise molecular mechanism remains ill defined. Here we demonstrate that different synaptotagmin isoforms (Syt I, II, and IV) are present in the same synaptic vesicle population from rat brain cortex. In addition, Syt I and II co-localize on the same small synaptic vesicle (SSV), and they heterodimerize in the presence of calcium with a concentration dependence resembling that of the starting phase of SSV exocytosis (EC50 = 6 +/- 4 microM). The association between Syt I and Syt II was demonstrated by immunoprecipitation of the native proteins and the recombinant cytoplasmic domains and by using fluorescence resonance energy transfer (FRET). Although a subpopulation of SSV containing Syt I and IV can be isolated, these two isoforms do not show a calcium-dependent interaction. These results suggest that the self-association of synaptotagmins with different calcium binding features may create a variety of calcium sensors characterized by distinct calcium sensitivities. This combinatorial hypothesis predicts that the probability of a single SSV exocytic event is determined, in addition to the gating properties of the presynaptic calcium channels, by the repertoire and relative abundance of distinct synaptotagmin isoforms present on the SSV surface. PMID- 9867812 TI - Human thymine DNA glycosylase binds to apurinic sites in DNA but is displaced by human apurinic endonuclease 1. AB - In vitro, following the removal of thymine from a G.T mismatch, thymine DNA glycosylase binds tightly to the apurinic site it has formed. It can also bind to an apurinic site opposite S6-methylthioguanine (SMeG) or opposite any of the remaining natural DNA bases. It will therefore bind to apurinic sites formed by spontaneous depurination, chemical attack, or other glycosylases. In the absence of magnesium, the rate of dissociation of the glycosylase from such complexes is so slow (koff 1.8 - 3.6 x 10(-5) s-1; i.e. half-life between 5 and 10 h) that each molecule of glycosylase removes essentially only one molecule of thymine. In the presence of magnesium, the dissociation rates of the complexes with C.AP and SMeG.AP are increased more than 20-fold, allowing each thymine DNA glycosylase to remove more than one uracil or thymine from C.U and SMeG.T mismatches in DNA. In contrast, magnesium does not increase the dissociation of thymine DNA glycosylase from G.AP sites sufficiently to allow it to remove more than one thymine from G.T mismatches. The bound thymine DNA glycosylase prevents human apurinic endonuclease 1 (HAP1) cutting the apurinic site, so unless the glycosylase was displaced, the repair of apurinic sites would be very slow. However, HAP1 significantly increases the rate of dissociation of thymine DNA glycosylase from apurinic sites, presumably through direct interaction with the bound glycosylase. This effect is concentration-dependent and at the probable normal concentration of HAP1 in cells the dissociation would be fast. This interaction couples the first step in base excision repair, the glycosylase, to the second step, the apurinic endonuclease. The other proteins involved in base excision repair, polymerase beta, XRCC1, and DNA ligase III, do not affect the dissociation of thymine DNA glycosylase from the apurinic site. PMID- 9867813 TI - Calcium influx activates extracellular-regulated kinase/mitogen-activated protein kinase pathway through a calmodulin-sensitive mechanism in PC12 cells. AB - Evidence suggests that membrane depolarization is able to promote neuronal survival through a sustained, although moderate, increase in the intracellular calcium. We have used the PC12 cell line to study the possible intracellular pathways that can be activated by calcium influx. Previously, we observed that membrane depolarization-induced calcium influx was able to activate the extracellular-regulated kinase (ERK)/mitogen-activated protein kinase pathway and most of this activation was calmodulin-dependent. We demonstrated that a part of the ERK activation is due to the phosphorylation of the epidermal growth factor receptor. Here, we show that both the epidermal growth factor receptor phosphorylation and the Shc-Grb2-Ras activation are not calmodulin-modulated. Moreover, dominant negative mutant Ha-ras (Asn-17) prevents the activation on ERKs by membrane depolarization, suggesting that Ras and calmodulin are both necessaries to activate ERKs by membrane depolarization. We failed to observe any significant induction and/or modulation of the A-Raf, B-Raf or c-Raf-1 kinase activities, thus suggesting the existence of a MEK kinase different from the classical Raf kinases that directly or indirectly can be modulated by Ca2+/calmodulin. PMID- 9867814 TI - A mechanism for Tn5 inhibition. carboxyl-terminal dimerization. AB - Tn5 is unique among prokaryotic transposable elements in that it encodes a special inhibitor protein identical to the Tn5 transposase except lacking a short NH2-terminal DNA binding sequence. This protein regulates transposition through nonproductive protein-protein interactions with transposase. We have studied the mechanism of Tn5 inhibition in vitro and find that a heterodimeric complex between the inhibitor and transposase is critical for inhibition, probably via a DNA-bound form of transposase. Two dimerization domains are known in the inhibitor/transposase shared sequence, and we show that the COOH-terminal domain is necessary for inhibition, correlating with the ability of the inhibitor protein to homodimerize via this domain. This regulatory complex may provide clues to the structures of functional synaptic complexes. Additionally, we find that NH2- and COOH-terminal regions of transposase or inhibitor are in functional contact. The NH2 terminus appears to occlude transposase homodimerization (hypothetically mediated by the COOH terminus), an effect that might contribute to productive transposition. Conversely, a deletion of the COOH terminus uncovers a secondary DNA binding region in the inhibitor protein which is probably located near the NH2 terminus. PMID- 9867815 TI - Characterization of calcineurin in human neutrophils. Inhibitory effect of hydrogen peroxide on its enzyme activity and on NF-kappaB DNA binding. AB - We describe here a specific calcineurin activity in neutrophil lysates, which is dependent on Ca2+, inhibited by trifluoroperazine, and insensitive to okadaic acid. Immunoblotting experiments using a specific antiserum recognized both the A and B chains of calcineurin. Neutrophils treated with cyclosporin A or FK 506 showed a dose-dependent inhibition of calcineurin activity. The effect of oxidant compounds on calcineurin activity was also investigated. Neutrophils treated with hydrogen peroxide (H2O2), where catalase was inhibited with aminotriazole, exhibited a specific inhibition of calcineurin activity. However, the addition of reducing agents to neutrophil extracts partially reversed the inhibition caused by H2O2. A similar inhibitory effect of H2O2 on calcineurin activity was observed to occur in isolated lymphocytes. This is the first demonstration that redox agents modulate calcineurin activity in a cellular system. In addition, electrophoretic mobility shift assays revealed that lipopolysaccharide-induced activation of NF-kappaB in human neutrophils is inhibited by cell pretreatment with H2O2 in a dose-dependent manner. These data indicate that calcineurin activity regulates the functional activity of lipopolysaccharide-induced NF kappaB/Rel proteins in human neutrophils. These data indicate a role of peroxides in the modulation of calcineurin activity and that the H2O2-dependent NF-kappaB inactivation in neutrophils occurs in concert with inhibition of calcineurin. PMID- 9867816 TI - Sequential cleavage and excision of a segment of the thyrotropin receptor ectodomain. AB - The thyrotropin (TSH) receptor belongs to a subfamily of G protein-coupled receptors, which also includes luteinizing hormone and follicle-stimulating hormone receptors. The TSH receptor (TSHR) differs from the latter by the presence of an additional specific segment in the C-terminal part of its ectodomain. We show here that this insertion is excised in the majority of receptor molecules. Preparation of specific monoclonal antibodies to this region, microsequencing, enzyme-linked immunosorbent assay, and immunoblot studies have provided insight into the mechanisms of this excision. In the human thyroid gland, N termini of the transmembrane receptor beta subunit were found to be phenylalanine 366 and leucines 370 and 378. In transfected L cells a variety of other more proximal N termini were found, probably corresponding to incomplete excisions. The most extreme N terminus was observed to lie at Ser-314. These observations suggest that after initial cleavage at Ser-314 the inserted fragment of TSHR is progressively clipped out by a series of cleavage reactions progressing up to amino acids 366-378. The impossibility of recovering the excised fragment from purified receptor, cell membranes, or culture medium supports this interpretation. The cleavage enzyme has previously been shown to be inhibited by BB-2116, an inhibitor of matrix metalloproteases. However, we show here that it is unaffected by tissue inhibitors of metalloproteases. The cleavage enzyme is very similar to TACE (tumor necrosis factor alpha-converting enzyme) in both these characteristics. However, incubation of the TSH receptor with the purified recombinant catalytic domain of TACE, co-transfection of cells with TACE and TSHR expression vectors, and the use of mutated Chinese hamster ovary cells in which TACE is inactive suggested that the TSHR cleavage enzyme is different from TACE. TACE and TSHR cleavage enzyme may thus possibly be related but different members of the adamalysin family of metzincin metalloproteases. PMID- 9867817 TI - Mutations in a peptidylprolyl-cis/trans-isomerase gene lead to a defect in 3'-end formation of a pre-mRNA in Saccharomyces cerevisiae. AB - In a genetic screen aimed at the identification of trans-acting factors involved in mRNA 3'-end processing of budding yeast, we have previously isolated two temperature-sensitive mutants with an apparent defect in the 3'-end formation of a plasmid-derived pre-mRNA. Surprisingly, both mutants were rescued by the essential gene ESS1/PTF1 that encoded a putative peptidylprolyl-cis/trans isomerase (PPIase) (Hani, J., Stumpf, G., and Domdey, H. (1995) FEBS Lett. 365, 198-202). Such enzymes, which catalyze the cis/trans-interconversion of peptide bonds N-terminal of prolines, are suggested to play a role in protein folding or trafficking. Here we report that Ptf1p shows PPIase activity in vitro, displaying an unusual substrate specificity for peptides with phosphorylated serine and threonine residues preceding proline. Both mutations were found to result in amino acid substitutions of highly conserved residues within the PPIase domain, causing a marked decrease in PPIase activity of the mutant enzymes. Our results are suggestive of a so far unknown involvement of a PPIase in mRNA 3'-end formation in Saccharomyces cerevisiae. PMID- 9867819 TI - Denatured states of tick anticoagulant peptide. Compositional analysis of unfolded scrambled isomers. AB - In the presence of denaturant and thiol catalyst, a disulfide-containing protein denatures and converts to a mixture of scrambled isomers, which can be purified and structurally characterized. Scrambled isomers adopt a different conformation and a varied extent of unfolding. Their relative concentration (composition) signals the state of unfolding of the denatured protein and is determined by the denaturing condition. In this report, tick anticoagulant peptide (TAP) (60 amino acids and 3 disulfides) has been denatured in the presence of urea, guanidine hydrochloride, guanidine thiocyanate, organic solvents, and at elevated temperature. The recoveries of scrambled TAP were analyzed. The results demonstrate that each denaturing condition generates a unique structure (composition of scrambled species) of denatured TAP. Among various species of scrambled TAP, the beads-form species contains the smallest disulfide loop and appears to represent the most extensively unfolded state. The yield of the beads form species as a fraction of the total denatured TAP is invariably determined by the strength of the denaturing condition. PMID- 9867818 TI - Conformational change of the catalytic subunit of glucose-6-phosphatase in rat liver during the fetal-to-neonatal transition. AB - The glucose-6-phosphatase system was investigated in fetal rat liver microsomal vesicles. Several observations indicate that the orientation of the catalytic subunit is different in the fetal liver in comparison with the adult form: (i) the phosphohydrolase activity was not latent using glucose-6-phosphate as substrate, and in the case of other phosphoesters it was less latent; (ii) the intravesicular accumulation of glucose upon glucose-6-phosphate hydrolysis was lower; (iii) the size of the intravesicular glucose-6-phosphate pool was independent of the glucose-6-phosphatase activities; (iv) antibody against the loop containing the proposed catalytic site of the enzyme inhibited the phosphohydrolase activity in fetal but not in adult rat liver microsomes. Glucose 6-phosphate, phosphate, and glucose uptake could be detected by both light scattering and/or rapid filtration method in fetal liver microsomes; however, the intravesicular glucose-6-phosphate and glucose accessible spaces were proportionally smaller than in adult rat liver microsomes. These data demonstrate that the components of the glucose-6-phosphatase system are already present, although to a lower extent, in fetal liver, but they are functionally uncoupled by the extravesicular orientation of the catalytic subunit. PMID- 9867820 TI - Dishevelled proteins lead to two signaling pathways. Regulation of LEF-1 and c Jun N-terminal kinase in mammalian cells. AB - Dishevelled (Dsh/Dvl) proteins are known to mediate Wnt signaling by up regulating beta-catenin levels and stimulating T cell factor (TCF)/LEF-1 dependent transcription. We have identified a new Dvl-mediated signaling pathway in that mouse Dvl proteins, when expressed in COS-7 cells, stimulate c-Jun dependent transcription activity and the kinase activity of the c-Jun N-terminal kinase (JNK). The DEP domain of Dvl1 is essential for JNK activation. By contrast, all three conserved domains of Dvl, including DIX, PDZ, and DEP, are required for up-regulation of beta-catenin and for stimulation of LEF-1-mediated transcription in mammalian cells. Thus, Dvl can lead to two different signaling pathways. Furthermore, the small G proteins of Cdc42 or Rac1, which are involved in JNK activation by many stimuli, do not appear to play a major role in Dvl mediated JNK activation, because the dominant negative mutants of Cdc42 and Rac1 could not inhibit Dvl-induced JNK activation. This suggests that Dvl may activate JNK via novel pathways. PMID- 9867821 TI - Cooperative role for activated alpha4 beta1 integrin and chondroitin sulfate proteoglycans in cell adhesion to the heparin III domain of fibronectin. Identification of a novel heparin and cell binding sequence in repeat III5. AB - We recently reported that the heparin (Hep) III domain of fibronectin contains the H2 cell adhesion site in repeat III5 which binds activated alpha4 integrins. We have now further characterized the heparin and cell binding activities of this domain. A recombinant fragment containing repeats III4-III5 (FN-III4-5) induced Jurkat cell adhesion upon integrin activation with Mn2+ or TS2/16 monoclonal antibody (anti-beta1). Adhesion of Mn2+-treated cells to FN-III4-5 or FN-III5 fragments was inhibited by chondroitinase ABC and ACII but not by the anti-alpha4 monoclonal antibody HP2/1. In contrast, HP2/1 completely blocked adhesion of TS2/16-treated cells while chondroitinase had a partial (FN-III4-5) or minor (FN III5) effect. Thus, the role of each receptor depended on the stimulus used to activate alpha4 beta1. The combination of HP2/1 and chondroitinase at dilutions which did not inhibit when used individually abolished adhesion of Mn2+ or TS2/16 treated cells to both fragments, indicating a cooperative effect between alpha4beta1 and chondroitin sulfate proteoglycans (CSPG). Furthermore, we have identified a 20-amino acid sequence in III5 (HBP/III5) which binds heparin and induces cell adhesion via CSPG exclusively. Although soluble HBP/III5 was a poor inhibitor, when combined with H2, it abolished adhesion to FN-III4-5 and FN-III5 fragments. These results establish that adhesion to the Hep III domain involves the cooperation of activated alpha4 beta1 and CSPG and show that HBP/III5 is a novel heparin and CSPG-binding site contributing to cell adhesion to this domain. PMID- 9867822 TI - Fluid shear stress stimulates big mitogen-activated protein kinase 1 (BMK1) activity in endothelial cells. Dependence on tyrosine kinases and intracellular calcium. AB - Mitogen-activated protein (MAP) kinases including ERK1/2 and JNK play an important role in shear stress-mediated gene expression in endothelial cells (EC). A new MAP kinase termed big MAP kinase 1 (BMK1/ERK5) has been shown to phosphorylate and activate the transcription factor MEF2C, which is highly expressed in EC. To determine the effects of shear stress on BMK1, bovine aortic EC were exposed to steady laminar flow (shear stress = 12 dynes/cm2). Flow activated BMK1 within 10 min with peak activation at 60 min (7.1 +/- 0.6-fold) in a force-dependent manner. Flow was the most powerful activator of BMK1, significantly greater than H2O2 or sorbitol. An important role for non-Src tyrosine kinases in flow-mediated BMK1 activation was demonstrated by inhibition with herbimycin A, but not with the Src inhibitor PP1 or overexpression of kinase inactive c-Src. BMK1 activation was calcium-dependent as shown by inhibition with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxymethyl ester or thapsigargin. As shown by specific inhibitors or activators, flow-mediated BMK1 activation was not regulated by the following: intracellular redox state; intracellular NO; protein kinase A, C, or G; calcium/calmodulin-dependent kinase; phosphatidylinositol 3-kinase; or arachidonic acid metabolism. In summary, flow potently stimulates BMK1 in EC by a mechanism dependent on a tyrosine kinase(s) and calcium mobilization, but not on c-Src, redox state, or NO production. PMID- 9867823 TI - Turnover analysis of glutamate receptors identifies a rapidly degraded pool of the N-methyl-D-aspartate receptor subunit, NR1, in cultured cerebellar granule cells. AB - The number, composition, and location of receptors in neurons are critically important factors in determining the neuron's response to neurotransmitters. The functional expression of receptors appears to be regulated both generally, at the level of transcription or translation, and locally, at the level of the individual synapse. A key component in the regulation of any protein is its turnover rate, which, measured in half-lives, ranges from a few minutes to several days. In the present study, we measured the turnover rates of subunits of N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors, the two major ionotropic glutamate receptors, using cultured cerebellar granule cells. Turnover rates for NR1, NR2A/B, GluR2/3, and GluR4 subunits were determined by pulse-chase labeling of cells with [35S]methionine. Half-lives were found to be 18 +/- 5 h and 23 +/- 8 h for the AMPA receptor subunits GluR2/3 and GluR4, respectively, and 16 +/- 5 h for NR2A. The NR1 subunit showed a biphasic decay with half-lives of 2 and 34 h for the rapidly and slowly degraded populations, respectively. Splice variants of the NR1 subunit with different carboxyl-terminal cassettes, C2 and C2', showed similar biphasic degradation patterns. To further characterize the rapidly degraded pool of NR1, surface receptors were labeled by biotinylation, and half-lives of the biotinylated proteins were determined. All surface NR1 was slowly degraded with a pattern similar to that of NR2A, GluR2/3, and GluR4, suggesting that the rapidly degraded pool is confined to the cytoplasm and not assembled with NR2 subunits. A significant amount of NR1 was not immunoprecipitated by NR2 subunit-specific antibodies after solubilization with deoxycholate. This unassembled pool, but not the assembled one, was greatly diminished following treatment of cycloheximide for 5 h, indicating that the rapidly degraded pool of NR1 is not assembled with NR2. These results show that NMDA and AMPA receptors have similar turnover rates, but NMDA receptors have a separate pool of NR1 subunits that is rapidly degraded and accounts for most of the intracellular pool. PMID- 9867824 TI - UV irradiation activates JNK and increases alphaI(I) collagen gene expression in rat hepatic stellate cells. AB - Hepatic stellate cells (HSCs) become activated into myofibroblast-like cells during the early stages of hepatic injury associated with fibrogenesis. The subsequent dysregulation of alphaI(I) collagen gene expression is a central pathogenetic step during the development of cirrhosis. Our recent study in rat HSCs (Davis, B. H., Chen, A., and Beno, D. (1996) J. Biol. Chem. 271, 11039 11042) found that ERK1,2 activation might be required for maximal alphaI(I) collagen gene expression. However, the role of the parallel JNK cascade in regulating alphaI(I) collagen gene expression was unknown. In this study, we initially found that UV irradiation of HSCs activated JNK but not ERK1,2. Furthermore, UV irradiation increased endogenous alpha I(I) collagen mRNA abundance and stimulated alpha I(I) collagen gene transcription in HSCs. The effect of the activation of JNK and Jun on alpha I(I) collagen gene expression was further evaluated via transfection of chloramphenicol acetyltransferase reporter plasmids with various sizes of truncated 5' upstream promoter sequence (UPS) of the alphaI(I) collagen gene. This revealed that dominant negative transcription factor JUN suppressed alpha I(I) collagen gene transcription in HSCs maintained in media with 20% serum and constitutively activated JUN increased alphaI(I) collagen gene transcription in HSCs cultured in media with 0.4% serum. UV activated JNK utilized a distal GC box in the 5'-UPS of the collagen gene to regulate gene transcription. This observation was confirmed by site-directed mutagenesis. In co-transfection experiments, the col chloramphenicol acetyltransferase reporter with a mutagenized GC box was not suppressed by dn-JUN and was not stimulated by activated JUN or by UV irradiation. Southwestern blotting analyses and gel shift assays with basic transcription element-binding protein antiserum suggested that the GC box was bound by basic transcription element-binding protein, a recently described DNA binding protein. In conclusion, the current study combined with our previous report suggests that ERK1,2 and JNK cascades regulate alphaI(I) collagen expression in HSCs through different regions of the 5'-UPS of the gene. The distal GC box in the 5'-UPS of the alphaI(I) collagen gene may play a central role in receiving extracellular signals through the JNK pathway. PMID- 9867825 TI - The kidney-expressed winged helix transcription factor FREAC-4 is regulated by Ets-1. A possible role in kidney development. AB - In this paper we show that the kidney-expressed winged helix transcription factor FREAC-4 is regulated by Ets-1, another kidney-expressed transcription factor. Through transfection experiments three Ets-1 cis-elements are identified within the first 152 nucleotides upstream of the transcription start in the freac-4 promoter. These sites are confirmed in a DNase I in vitro protection assay using recombinant Ets-1 protein. In cotransfection experiments using an Ets-1 expression vector, the induction of freac-4 reporter gene activity is attenuated approximately 6-fold when the three Ets-1 binding sites are mutated. Furthermore, we demonstrate that overexpression of Ets-1 in the human embryonic kidney cell line 293 is sufficient to increase freac-4 mRNA levels. These results are compatible with the hypothesis that Ets-1 acts as an upstream regulator of FREAC 4 expression during kidney development. PMID- 9867826 TI - beta-lactoglobulin binds palmitate within its central cavity. AB - Bovine beta-lactoglobulin (beta-Lg) has been studied extensively in both the isolated and the naturally occurring states. It is a commercially important whey protein of obvious nutritional value but, so far, one that has no clearly identified biological function. In common with many of the other members of the lipocalin family to which it belongs, beta-Lg binds hydrophobic ligands, and it appears possible that there are at least two distinct binding sites per monomer for a variety of ligands. By comparison with other members of the family, there is a probable binding site in the central cavity of the molecule that is formed by the eight antiparallel beta-strands that are typical of the lipocalins. We have now cocrystallized beta-Lg with palmitic acid, and the refined structure (R = 0.204, Rfree = 0.240 for 6,888 reflections to 2.5-A resolution) reveals that the ligand binds in the central cavity in a manner similar to the binding of retinol to the related lipocalin, serum retinol-binding protein. The carboxyl group binds to both Lys-60 and Lys-69 at the entrance to the cavity. The hydrophobic tail stretches in an almost fully extended conformation into the center of the protein. This is the first direct observation of a ligand binding to beta-Lg. PMID- 9867827 TI - Calcium influx factor, further evidence it is 5, 6-epoxyeicosatrienoic acid. AB - We present evidence in astrocytes that 5,6-epoxyeicosatrienoic acid, a cytochrome P450 epoxygenase metabolite of arachidonic acid, may be a component of calcium influx factor, the elusive link between release of Ca2+ from intracellular stores and capacitative Ca2+ influx. Capacitative influx of extracellular Ca2+ was inhibited by blockade of the two critical steps in epoxyeicosatrienoic acid synthesis: release of arachidonic acid from phospholipid stores by cytosolic phospholipase A2 and cytochrome P450 metabolism of arachidonic acid. AAOCF3, which inhibits cytosolic phospholipase A2, blocked thapsigargin-stimulated release of arachidonic acid as well as thapsigargin-stimulated elevation of intracellular free calcium. Inhibition of P450 arachidonic acid metabolism with SKF525A, econazole, or N-methylsulfonyl-6-(2-propargyloxyphenyl)hexanamide, a substrate inhibitor of P450 arachidonic acid metabolism, also blocked thapsigargin-stimulated Ca2+ influx. Nano- to picomolar 5, 6-epoxyeicosatrienoic acid induced [Ca2+]i elevation consistent with capacitative Ca2+ influx. We have previously shown that 5, 6-epoxyeicosatrienoic acid is synthesized and released by astrocytes. When 5,6-epoxyeicosatrienoic acid was applied to the rat brain surface, it induced vasodilation, suggesting that calcium influx factor may also serve a paracrine function. In summary, our results suggest that 5,6 epoxyeicosatrienoic acid may be a component of calcium influx factor and may participate in regulation of cerebral vascular tone. PMID- 9867828 TI - Analysis of the MRP8-MRP14 protein-protein interaction by the two-hybrid system suggests a prominent role of the C-terminal domain of S100 proteins in dimer formation. AB - Calcium-binding S100 proteins are thought to play a central role in calcium mediated signal transduction pathways. They consist of two helix-loop-helix, calcium-binding EF-hand domains. A characteristic feature is their tendency to form homo- and/or heterodimeric complexes. This report presents for the first time a functional "in vivo" approach to the analysis of S100 protein dimerization. Using the two-hybrid system we analyzed the dimerization of MRP8 (S100A8) and MRP14 (S100A9), two S100 proteins expressed in myeloid cells. It is reported that the MRP8-MRP14 heteromer is the clearly preferred complex in both man and mouse. The ability to homodimerize, however, appears to be restricted to the murine MRPs. Interaction analysis of chimeric murine/human MRP14 proteins indicates, that the C-terminal EF-hand domain plays a prominent role in MRP8 MRP14 interaction and determines the specificity of dimerization. Site-directed mutagenesis of four evolutionary conserved hydrophobic amino acids, which have been recently supposed to be essential for S100 protein dimerization, suggests that at least one of these, namely the most N-terminal located residue, is not critical for dimerization. PMID- 9867829 TI - Evidence for cysteine persulfide as reaction product of L-Cyst(e)ine C-S-lyase (C DES) from Synechocystis. Analyses using cystine analogues and recombinant C-DES. AB - The pyridoxal phosphate-dependent monomeric L-cysteine/cystine C-S-lyase (C-DES), previously isolated from Synechocystis PCC 6714 by its capacity to direct [2Fe 2S] cluster assembly of ferredoxin in vitro (Leibrecht, I., and Kessler, D. (1997) J. Biol. Chem. 272, 10442-10447), has now been cloned, sequenced, and overexpressed in Escherichia coli. The amino acid sequence of C-DES was found to be nearly identical (92% identity) to the open reading frame slr2143 of Synechocystis PCC 6803 and showed a more distant relationship to the NifS family of proteins (about 27% identity). Recombinant C-DES displayed activities equal to the isolate from Synechocystis in terms of the cyst(e)ine lyase reaction and holoferredoxin formation which recommended its use for functional and mechanistic studies. Investigation of the substrate spectrum for beta-elimination found L cysteine to be a poor substrate (kcat approximately 0.15 s-1) in contrast to L cystine (kcat = 36 s-1) and several related compounds. Of these compounds, desaminocystine (S-(carboxyethylthio)-L-cysteine) was used for C-DES-mediated persulfide generation. Stabilization of the linear persulfide 3-(disulfanyl) propionic acid was achieved by cyclization as a novel intramolecular trapping reaction; this yielded 1,2-dithiolan-3-one which was isolated and identified by chemical analyses. PMID- 9867830 TI - The association of initiation factor 4F with poly(A)-binding protein is enhanced in serum-stimulated Xenopus kidney cells. AB - Serum stimulation of cultured Xenopus kidney cells results in enhanced phosphorylation of the translational initiation factor (eIF) 4E and promotes a 2.8-fold increase in the binding of the adapter protein eIF4G to eIF4E, to form the functional initiation factor complex eIF4F. Here we demonstrate the serum stimulated co-isolation of the poly(A)-binding protein (PABP) with the eIF4F complex. This apparent interaction of PABP with eIF4F suggests that a mechanism shown to be important in the control of translation in the yeast Saccharomyces cerevisiae also operates in vertebrate cells. We also present evidence that the signaling pathways modulating eIF4E phosphorylation and function in Xenopus kidney cells differ from those in several mammalian cell types studied previously. Experiments with the immunosuppressant rapamycin suggest that the mTOR signaling pathway is involved in serum-promoted eIF4E phosphorylation and association with eIF4G. Moreover, we could find little evidence for regulation of eIF4E function via interaction with the specific binding proteins 4E-BP1 or 4E BP2 in these cells. Although rapamycin abrogated serum-enhanced rates of protein synthesis and the interaction of eIF4G with eIF4E, it did not prevent the increase in association of eIF4G with PABP. This suggests that serum stimulates the interaction between eIF4G and PABP by a distinct mechanism that is independent of both the mTOR pathway and the enhanced association of eIF4G with eIF4E. PMID- 9867831 TI - The Tup1-Cyc8 protein complex can shift from a transcriptional co-repressor to a transcriptional co-activator. AB - Cyc8(Ssn6)-Tup1, a general co-repressor complex, is recruited to promoter DNA via interactions with DNA-binding regulatory proteins and inhibits the transcription of many different yeast genes. Previous studies have established that repression function of the complex is performed by one subunit of the complex, the Tup1 protein, and requires specific components of the RNA polymerase II holoenzyme such as Sin4 and Rgr1. In this study we test the transcriptional activity of the Cyc8 subunit using a lexA operator-containing reporter. We show that a LexA-Cyc8 hybrid stimulates transcription when expressed in a tup1Delta, a sin4Delta, or a rgr1Delta strain, suggesting that transcriptional activation is an intrinsic property of the Cyc8-Tup1 co-repressor. In support of this notion we demonstrate that Cyc8-Tup1 has a dual function on CIT2, a gene encoding a citrate synthase that is expressed upon mitochondrial dysfunction. First, we show that Cyc8-Tup1 is tethered to CIT2 promoter by interacting with the activation domain of Rtg3, a bHLH/L-Zip DNA-binding transactivator of CIT2. Next we demonstrate that Cyc8-Tup1 activates CIT2 transcription in response to mitochondrial dysfunction, and this stimulatory effect is mediated by Cyc8. In contrast, basal (noninduced) expression of this gene is inhibited by Tup1. These findings establish a positive role for the Cyc8-Tup1 complex in transcription and support a model by which specific metabolic signals may convert the Cyc8-Tup1 transcriptional co-repressor to a co-activator of certain promoters. PMID- 9867832 TI - Role of CCAAT enhancer-binding protein beta in the thyroid hormone and cAMP induction of phosphoenolpyruvate carboxykinase gene transcription. AB - Transcription of the gene for phosphoenolpyruvate carboxykinase (PEPCK) is stimulated by thyroid hormone (T3) and cAMP. Two DNA elements in the PEPCK promoter are required for T3 responsiveness including a thyroid hormone response element and a binding site called P3(I) for the CCAAT enhancer-binding protein (C/EBP). Both the alpha and beta isoforms of C/EBP are highly expressed in the liver. C/EBPalpha contributes to the liver-specific expression and cAMP responsiveness of the PEPCK gene. In this study, we examined the ability of C/EBPbeta when bound to the P3(I) site to regulate PEPCK gene expression. We report that C/EBPbeta can stimulate basal expression and participate in the induction of PEPCK gene transcription by T3 and cAMP. The cAMP-responsive element binding protein and AP1 proteins that contribute to the induction by cAMP are not involved in the stimulation by T3. A small region of the transactivation domain of C/EBPbeta is sufficient for the stimulation of basal expression and cAMP responsiveness. Our results suggest that C/EBPalpha and C/EBPbeta are functionally interchangeable when bound to the P3(I) site of the PEPCK promoter. PMID- 9867833 TI - Evidence for (Mac1p)2.DNA ternary complex formation in Mac1p-dependent transactivation at the CTR1 promoter. AB - The Mac1 protein in Saccharomyces cerevisiae is required for the expression CTR1 and FRE1, which, respectively, encode the copper permease and metal reductase that participate in copper uptake. Mac1p binds to a core GCTC sequence present as a repeated unit in the promoters of both genes. We show here that Mac1p DNA binding required an intact N-terminal protein domain that includes a likely zinc finger motif. This binding was enhanced by the presence of a TATTT sequence immediately 5' to the core GCTC, in contrast to a TTTTT one. This increased binding was demonstrated clearly in vitro in electrophoretic mobility shift assays that showed Mac1p.DNA complex formation to a single TATTTGCTC element but not to a TTTTTGCTC one. Furthermore, the fraction of Mac1p in a ternary (Mac1p)2.DNA complex in comparison to a binary Mac1p.DNA complex increased when the DNA included two TATTTGCTC elements. A similar increase in ternary complex formation was demonstrated upon homologous mutation of the FRE1 Mac1p-dependent promoter element. The in vivo importance of this ternary complex formation at the CTR1 promoter was indicated by the stronger trans-activity of this promoter mutated to contain two TATTT elements and the attenuated activity of a mutant promoter containing two TTTTT elements that in vitro supported only a weak ternary complex signal in the shift assay. The stronger binding to TATTT appeared due to a more favorable protein contact with adenine in comparison to thymine at this position. An in vivo two-hybrid analysis demonstrated a Mac1p-Mac1p protein protein interaction. This Mac1p-Mac1p interaction may promote (Mac1p)2.DNA ternary complex formation at Mac1p-responsive upstream activating sequences. PMID- 9867834 TI - Structure of two fragments of the third cytoplasmic loop of the rat angiotensin II AT1A receptor. Implications with respect to receptor activation and G-protein selection and coupling. AB - The structural bases that render the third intracellular loop (i3) of the rat angiotensin II AT1A receptor one of the cytoplasmic domains responsible for G protein coupling are still unknown. The three-dimensional structures of two overlapping peptides mapping the entire i3 loop and shown to differently interact with purified G-proteins have been obtained by simulated annealing calculations, using NMR-derived constraints collected in 70% water/30% trifluoroethanol solution. While the NH2-terminal half, Ni3, residues 213-231, adopts a stable amphipathic alpha-helix, extending over almost the entire peptide, a more flexible conformation is found for the COOH-terminal half, Ci3, residues 227-242. For this peptide, a cis-trans isomerization around the Lys6-Pro7 peptide bond generates two exchanging isomers adopting similar conformations, with an alpha helix spanning from Asn9 to Ile15 and a poorly defined NH2 terminus. A quite distinct structural organization is found for the sequence EIQKN, common to Ni3 and Ci3. The data do suggest that the extension and orientation of the amphipathic alpha-helix, present in the proximal part of i3, may be modulated by the distal part of the loop itself through the Pro233 residue. A molecular model where this possibility is considered as a mechanism for G-protein selection and coupling is presented. PMID- 9867835 TI - Connexin-43 hemichannels opened by metabolic inhibition. AB - The cause of altered ionic homeostasis leading to cell death during ischemia and metabolic inhibition is unclear. Hemichannels, which are precursors to gap junctions, are nonselective ion channels that are permeable to molecules of less than Mr 1000. We show that hemichannels open upon exposure to calcium-free solutions when they are either heterologously overexpressed in HEK293 cells or endogenously expressed in cardiac ventricular myocytes. In the presence of normal extracellular calcium, hemichannels open during metabolic inhibition. During ischemia and other forms of metabolic inhibition, activation of relatively few hemichannels will seriously compromise the cell's ability to maintain ionic homeostasis, which is an essential step promoting cell death. PMID- 9867837 TI - A new heat shock protein that binds nucleic acids. AB - We describe the isolation of Hsp15, a new, very abundant heat shock protein that binds to DNA and RNA. Hsp15 is well conserved and related to a number of RNA binding proteins, including ribosomal protein S4, RNA pseudouridine synthase, and tyrosyl-tRNA synthetase. The region shared between these proteins appears to represent a common, but previously unrecognized, RNA binding motif. Filter binding studies showed that Hsp15 binds to a 17-mer single-stranded RNA with a dissociation constant of 9 microM in 22.5 mM Hepes, pH 7. 0, 5 mM MgCl2. A role of Hsp15 in binding nucleic acids puts this protein into a different functional category from that of many other heat shock proteins that act as molecular chaperones or proteases on protein substrates. PMID- 9867836 TI - cDNA cloning, expression, and mutagenesis study of liver-type prostaglandin F synthase. AB - Prostaglandin (PG) F synthase catalyzes the reduction of PGD2 to 9alpha,11beta PGF2 and that of PGH2 to PGF2alpha on the same molecule. PGF synthase has at least two isoforms, the lung-type enzyme (Km value of 120 microM for PGD2 (Watanabe, K., Yoshida, R., Shimizu, T., and Hayaishi, O. (1985) J. Biol. Chem. 260, 7035-7041) and the liver-type one (Km value of 10 microM for PGD2 (Chen, L. Y., Watanabe, K., and Hayaishi, O. (1992) Arch. Biochem. Biophys. 296, 17-26)). The liver-type enzyme was presently found to consist of a 969-base pair open reading frame coding for a 323-amino acid polypeptide with a Mr of 36,742. Sequence analysis indicated that the bovine liver PGF synthase had 87, 79, 77, and 76% identity with the bovine lung PGF synthase and human liver dihydrodiol dehydrogenase (DD) isozymes DD1, DD2, and DD4, respectively. Moreover, the amino acid sequence of the liver-type PGF synthase was identical with that of bovine liver DD3. The liver-type PGF synthase was expressed in COS-7 cells, and its recombinant enzyme had almost the same properties as the native enzyme. Furthermore, to investigate the nature of catalysis and/or substrate binding of PGF synthase, we constructed and characterized various mutant enzymes as follows: R27E, R91Q, H170C, R223L, K225S, S301R, and N306Y. Although the reductase activities toward PGH2 and phenanthrenequinone (PQ) of almost all mutants were not inactivated, the Km values of R27E, R91Q, H170C, R223L, and N306Y for PGD2 were increased from 15 to 110, 145, 75, 180, and 100 microM, respectively, indicating that Arg27, Arg91, His170, Arg223, and Asn306 are essential to give a low Km value for PGD2 of the liver-type PGF synthase and that these amino acid residues serve in the binding of PGD2. Moreover, the R223L mutant among these seven mutants especially has a profound effect on kcat for PGD2 reduction. The Km values of R223L, K225S, and S301R for PQ were about 2-10-fold lower than the wild type value, indicating that the amino acid residues at 223, 225 and 301 serve in the binding of PQ to the enzyme. On the other hand, the Km value of H170C for PGH2 was 8-fold lower than that of the wild type, indicating that the amino acid residue at 170 is related to the binding of PGH2 to the enzyme and that Cys170 confer high affinity for PGH2. Additionally, the 5-fold increase in kcat/Km value of the N306Y mutant for PGH2 compared with the wild-type value suggests that the amino acid at 306 plays an important role in catalytic efficiency for PGH2. PMID- 9867838 TI - Caveolin interacts with Trk A and p75(NTR) and regulates neurotrophin signaling pathways. AB - Neurotrophins signal through Trk tyrosine kinase receptors and the low-affinity neurotrophin receptor p75(NTR). We have shown previously that activation of Trk A tyrosine kinase activity can inhibit p75(NTR)-dependent sphingomyelin hydrolysis, that caveolae are a localized site for p75(NTR) signaling, and that caveolin can directly interact with p75(NTR). The ability of caveolin to also interact with tyrosine kinase receptors and inhibit their activity led us to hypothesize that caveolin expression may modulate interactions between neurotrophin signaling pathways. PC12 cells were transfected with caveolin that was expressed efficiently and targeted to the appropriate membrane domains. Upon exposure to nerve growth factor (NGF), caveolin-PC12 cells were unable to develop extensive neuritic processes. Caveolin expression in PC12 cells was found to diminish the magnitude and duration of Trk A activation in vivo. This inhibition may be due to a direct interaction of caveolin with Trk A, because Trk A co-immunoprecipitated with caveolin from Cav-Trk A-PC12 cells, and a glutathione S-transferase-caveolin fusion protein bound to Trk A and inhibited NGF-induced autophosphorylation in vitro. Furthermore, the in vivo kinetics of the inhibition of Trk A tyrosine kinase activity by caveolin expression correlated with an increased ability of NGF to induce sphingomyelin hydrolysis through p75(NTR). In summary, our results suggest that the interaction of caveolin with neurotrophin receptors may have functional consequences in regulating signaling through p75(NTR) and Trk A in neuronal and glial cell populations. PMID- 9867839 TI - p38 mitogen-activated protein kinase regulates cyclooxygenase-2 mRNA stability and transcription in lipopolysaccharide-treated human monocytes. AB - p38 mitogen-activated protein kinase (MAPK) is activated by inflammatory stimuli such as bacterial lipopolysaccharide (LPS), interleukin-1, and tumor necrosis factor. We have previously shown that the pyridinyl imidazole SB 203580, which inhibits it, blocks the interleukin-1 induction of cyclooxygenase-2 (COX-2) and matrix metalloproteinase 1 and 3 mRNAs in fibroblasts. Here we explore the role of p38 MAPK in the response of human monocytes to LPS. 0.1 microM SB 203580 significantly inhibited the LPS induction of COX-2 and tumor necrosis factor protein and mRNAs. The activity of MAPK-activated protein kinase-2 (a substrate of p38 MAPK) in the cells was commensurately reduced. Some isoforms of c-jun N terminal kinase (which is also activated by LPS) are sensitive to SB 203580; the inhibitor had little effect on monocyte c-jun N-terminal kinases up to 2 microM. We investigated the mechanism of inhibition of COX-2 induction. Transcription (measured by a nuclear run-on assay) was 60% inhibited by SB 203580 (2 microM). Importantly, we found that p38 MAPK was essential for stabilizing COX-2 mRNA: when cells stimulated for 4 h with LPS were treated with actinomycin D, COX-2 mRNA decayed slowly. Treatment of stimulated cells with 2 microM SB 203580 caused a rapid disappearance of COX-2 mRNA, even with actinomycin D present. We conclude p38 MAPK plays a role in the transcription and stabilization of COX-2 mRNA. PMID- 9867840 TI - Identification of regulatory and catalytic domains in the apoptosis nuclease DFF40/CAD. AB - The DNA fragmentation factor (DFF) is composed of two subunits, the 40-kDa caspase-3-activated nuclease (DFF40/CAD) and its 45-kDa inhibitor (DFF45/ICAD). During apoptosis, DFF-40/CAD is activated by caspase-3-mediated cleavage of DFF45/ICAD. Mutational analysis of DFF40/CAD revealed that DFF40/CAD is composed of a C-terminal catalytic domain and an N-terminal regulatory domain. Deletion of the catalytic domain (residues 290-345) abrogated the caspase-3-induced nuclease activity of DFF40/CAD but not its ability to interact with DFF45/ICAD. Conversely, removal of the regulatory domain (residues 1-83) yielded a constitutively active DFF40/CAD nuclease that neither bound to its inhibitor nor required caspase-3 for activation. Amino acid alignment revealed that the regulatory domain of DFF40/CAD has homology to the N-terminal region of mammalian and Drosophila DFF45/ICAD and CIDE-N, a regulatory domain previously identified in pro-apoptotic CIDE proteins. Mutational analysis of the N-terminal region revealed mutants with diminished nuclease activity but with intact ability to bind DFF45/ICAD. Thus, CIDE-N represents a new type of domain that is associated with the regulation of the apoptosis/DNA fragmentation pathway. PMID- 9867841 TI - Analysis of a structural determinant in thrombin-protease nexin 1 complexes that mediates clearance by the low density lipoprotein receptor-related protein. AB - We recently identified a synthetic peptide, Pro47-Ile58, derived from the mature protease nexin 1 (PN1) sequence, that inhibited the low density lipoprotein receptor-related protein (LRP)-mediated internalization of thrombin-PN1 (Th-PN1) complexes. Presently, we have analyzed this sequence in Th-PN1 complex catabolism using two independent approaches: 1) An antibody was generated against Pro47 Ile58, which inhibited complex degradation by 70% but had no effect on the binding of the complexes to cell surface heparins. This places the structural determinant in PN1 mediating complex internalization by the LRP outside of the heparin-binding site. 2) Site-directed genetic variants of PN1 with a single Ala substitution at His48, or two Ala substitutions, one at His48 and another at Asp49, were expressed in Sf9 insect cells. The catabolic rate of complexes formed between Th and the singly substituted and doubly substituted variants was lowered to 50 and 15%, respectively, when compared with the catabolic rate of native Th PN1 complexes. This is the first analysis of a structural determinant in a serine protease inhibitor (SERPIN) required for LRP-mediated internalization and in part may explain the cryptic nature of this site in the unreacted serine protease inhibitor. PMID- 9867842 TI - Modulation of pancreatic acinar cell to cell coupling during ACh-evoked changes in cytosolic Ca2+. AB - The temporal changes in cytosolic free Ca2+ ([Ca2+]i), Ca2+-dependent membrane currents (Im), and gap junctional current (Ij) elicited by acetylcholine (ACh) were measured in rat pancreatic acinar cells using digital imaging and dual perforated patch-clamp recording. ACh (50 nM-5 microM) increased [Ca2+]i and evoked Im currents without altering Ij in 19 of 37 acinar cell pairs. Although [Ca2+]i rose asynchronously in cells comprising a cluster, the delay of the [Ca2+]i responses decreased with increasing ACh concentrations. Perfusion of inositol 1,4,5-trisphosphate (IP3) into one cell of a cluster resulted in [Ca2+]i responses in neighboring cells that were not necessarily in direct contact with the stimulated one. This suggests that extensive coupling between acinar cells provides a pathway for cell-to-cell diffusion of Ca2+-releasing signals. Strikingly, maximal (1-5 microM) ACh concentrations reduced Ij by 69 +/- 15% (n = 9) in 25% of the cell pairs subjected to dual patch-clamping. This decrease occurred shortly after the Im peak and was prevented by incubating acinar cells in a Ca2+-free medium, suggesting that uncoupling was subsequent to the initiation of the Ca2+-mobilizing responses. Depletion of Ca2+-sequestering stores by thapsigargin resulted in a reduction of intercellular communication similar to that observed with ACh. In addition, ACh-induced uncoupling was prevented by blocking nitric oxide production with L-nitro-arginine and restored by exposing acinar cells to dibutyryl cGMP. The results suggest that ACh-induced uncoupling and capacitative Ca2+ entry are regulated concurrently. Closure of gap junction channels may occur to functionally isolate nearby cells differing in their intrinsic sensitivity to ACh and thereby to allow for sustained activity of groups of secreting cells. PMID- 9867843 TI - Expression of three Caenorhabditis elegans N-acetylglucosaminyltransferase I genes during development. AB - UDP-N-acetylglucosamine:alpha-3-D-mannoside beta-1, 2-N acetylglucosaminyltransferase I (GnT I) is a key enzyme in the synthesis of Asn linked complex and hybrid glycans. Studies on mice with a null mutation in the GnT I gene have indicated that N-glycans play critical roles in mammalian morphogenesis. This paper presents studies on N-glycans during the development of the nematode Caenorhabditis elegans. We have cloned cDNAs for three predicted C. elegans genes homologous to mammalian GnT I (designated gly-12, gly-13, and gly 14). All three cDNAs encode proteins (467, 449, and 437 amino acids, respectively) with the domain structure typical of previously cloned Golgi-type glycosyltransferases. Expression in both insect cells and transgenic worms showed that gly-12 and gly-14, but not gly-13, encode active GnT I. All three genes were expressed throughout worm development (embryo, larval stages L1-L4, and adult worms). The gly-12 and gly-13 promoters were expressed from embryogenesis to adulthood in many tissues. The gly-14 promoter was expressed only in gut cells from L1 to adult developmental stages. Transgenic worms that overexpress any one of the three genes show no obvious phenotypic defects. The data indicate that C. elegans is a suitable model for further study of the role of complex N-glycans in development. PMID- 9867844 TI - Structural characterization of inter-alpha-inhibitor. Evidence for an extended shape. AB - Inter-alpha-inhibitor (IalphaI) is a 180-kDa serum protein consisting of three polypeptides. Two of these, the heavy chains 1 and 2 (H1 and H2), are of 75-80 kDa and have similar amino acid sequences. The third polypeptide, bikunin, has a molecular mass of 25 kDa and contains a 7-kDa chondroitin sulfate chain that is covalently linked to the C-terminal amino acid residues of H1 and H2. IalphaI has been shown to be required for the formation of the hyaluronan-containing extracellular matrix of certain cell types. How IalphaI exerts this function is not known, but it appears that upon interaction with cells, the heavy chains are released and become covalently linked to hyaluronan. Our results indicate that IalphaI and its heavy chains are extended molecules; thus, upon electron microscopy, IalphaI appeared to consist of two globular domains connected by a thin structure 31-nm long and the isolated heavy chains of a globular domain and a "tail" about 15-nm long. Analysis of the heavy chains by partial proteolysis showed that the C-terminal halves are particularly sensitive to hydrolysis indicating that they are loosely folded. Furthermore, electron microscopy showed that partially degraded heavy chains lacked the extended regions. Taken together, these results suggest that the N-terminal half of the heavy chains forms a globular domain, whereas the other half has an extended and loosely folded structure. PMID- 9867846 TI - Trypsinized cerebellar inositol 1,4,5-trisphosphate receptor. Structural and functional coupling of cleaved ligand binding and channel domains. AB - The type 1 inositol 1,4,5-trisphosphate receptor (IP3R1) is a tetrameric intracellular inositol 1,4,5-trisphosphate (IP3)-gated Ca2+ release channel (calculated molecular mass = approximately 313 kDa/subunit). We studied structural and functional coupling in this protein complex by limited (controlled) trypsinization of membrane fractions from mouse cerebellum, the predominant site for IP3R1. Mouse IP3R1 (mIP3R1) was trypsinized into five major fragments (I-V) that were positioned on the entire mIP3R1 sequence by immuno probing with 11 site-specific antibodies and by micro-sequencing of the N termini. Four fragments I-IV were derived from the N-terminal cytoplasmic region where the IP3-binding region extended over two fragments I (40/37 kDa) and II (64 kDa). The C-terminal fragment V (91 kDa) included the membrane-spanning channel region. All five fragments were pelleted by centrifugation as were membrane proteins. Furthermore, after solubilizing with 1% Triton X-100, all were co immunoprecipitated with the C terminus-specific monoclonal antibody that recognized only the fragment V. These data suggested that the native mIP3R1 channel is an assembly of four subunits, each of which is constituted by non covalent interactions of five major, well folded structural components I-V that are not susceptible to attack by mild trypsinolysis. Ca2+ release experiments further revealed that even the completely fragmented mIP3R1 retained significant IP3-induced Ca2+ release activity. These data suggest that structural coupling among five split components conducts functional coupling for IP3-induced Ca2+ release, despite the loss of peptide linkages. We propose structural-functional coupling in the mIP3R1, that is neighboring coupling between components I and II for IP3 binding and long-distant coupling between the IP3 binding region and the channel region (component V) beyond trypsinized gaps for ligand gating. PMID- 9867845 TI - Dual roles for glucokinase in glucose homeostasis as determined by liver and pancreatic beta cell-specific gene knock-outs using Cre recombinase. AB - Glucokinase (GK) gene mutations cause diabetes mellitus in both humans and mouse models, but the pathophysiological basis is only partially defined. We have used cre-loxP technology in combination with gene targeting to perform global, beta cell-, and hepatocyte-specific gene knock-outs of this enzyme in mice. Gene targeting was used to create a triple-loxed gk allele, which was converted by partial or total Cre-mediated recombination to a conditional allele lacking neomycin resistance, or to a null allele, respectively. beta cell- and hepatocyte specific expression of Cre was achieved using transgenes that contain either insulin or albumin promoter/enhancer sequences. By intercrossing the transgenic mice that express Cre in a cell-specific manner with mice containing a conditional gk allele, we obtained animals with either a beta cell or hepatocyte specific knock-out of GK. Animals either globally deficient in GK, or lacking GK just in beta cells, die within a few days of birth from severe diabetes. Mice that are heterozygous null for GK, either globally or just in the beta cell, survive but are moderately hyperglycemic. Mice that lack GK only in the liver are only mildly hyperglycemic but display pronounced defects in both glycogen synthesis and glucose turnover rates during a hyperglycemic clamp. Interestingly, hepatic GK knock-out mice also have impaired insulin secretion in response to glucose. These studies indicate that deficiencies in both beta cell and hepatic GK contribute to the hyperglycemia of MODY-2. PMID- 9867847 TI - Cooperative formation of the ligand-binding site of the inositol 1,4, 5 trisphosphate receptor by two separable domains. AB - Limited trypsin digestion of mouse cerebellar membrane fractions leads to fragmentation of the type 1 inositol 1,4,5-trisphosphate receptor (IP3R1) into five major components (Yoshikawa, F., Iwasaki, H., Michikawa, T., Furuichi, T., and Mikoshiba, K. (1999) J. Biol. Chem. 274, 316-327). Here we report that trypsin-fragmented mouse IP3R1 (mIP3R1) retains significant inositol 1,4,5 trisphosphate (IP3) binding activity that is comparable to the intact receptor in affinity, capacity, and specificity. This is despite the fact that the IP3 binding core (residues 226-578), which is close to the minimum for high affinity binding, is completely split into two tryptic fragments at the Arg-343 and/or Arg 345, around the center of the core. Furthermore, we have examined whether binding activity could be complemented in vitro by mixing two distinct glutathione S transferase (GST) fusion proteins, which were respectively composed of residues 1 343 and 341-604, almost corresponding to two split binding components, and separately expressed in Escherichia coli. The GST-fused residues 1-343 (GN) showed no binding affinity for IP3, whereas the GST-fused residues 341-604 (GC) displayed weak but definite activity with an affinity >100-fold lower than that of the native receptor. Upon mixing of both GN and GC, a high affinity site comparable to the native site appeared. We suggest that the IP3-binding pocket consists of two non-covalently but tightly associated structural domains each of which has a discrete function: the C-terminal domain alone has low affinity for IP3, whereas the N-terminal one alone is incapable of binding but is capable of potentiating binding affinity. PMID- 9867848 TI - The carboxyl-terminal region of biliary glycoprotein controls its tyrosine phosphorylation and association with protein-tyrosine phosphatases SHP-1 and SHP 2 in epithelial cells. AB - Biliary glycoprotein (Bgp, C-CAM, or CD66a) is an immunoglobulin-like cell adhesion molecule and functions as a tumor suppressor protein. We have previously shown that the Bgp1 isoform responsible for inhibition of colonic, liver, prostate, and breast tumor cell growth contains within its cytoplasmic domain two tyrosine residues positioned in immunoreceptor tyrosine-based inhibition motif (ITIM) consensus sequences. Moreover, we determined that these residues, upon phosphorylation, associate with the protein-tyrosine phosphatase SHP-1. In this report, we have further evaluated the structural bases of the association of Bgp1 with Tyr phosphatases. First, we demonstrate that Bgp1 also associates with the SHP-2 Tyr phosphatase, but not with an unrelated Tyr phosphatase, PTP-PEST. Association of Bgp1 and SHP-2 involves the Tyr residues within the Bgp1 ITIM sequences, Val at position +3 relative to the second Tyr (Tyr-515), and the SHP-2 N-terminal SH2 domain. In addition, our results indicate that residues +4, +5, and +6 relative to Tyr-515 in the Bgp1 cytoplasmic domain play a significant role in these interactions, as their deletion reduced Bgp1 Tyr phosphorylation and association with SHP-1 and SHP-2 by as much as 80%. Together, these results indicate that both SHP-1 and SHP-2 interact with the Bgp1 cytoplasmic domain via ITIM-like sequences. Furthermore, they reveal that the C-terminal amino acids of Bgp1 are critical for these interactions. PMID- 9867849 TI - The orphan nuclear receptor SHP inhibits agonist-dependent transcriptional activity of estrogen receptors ERalpha and ERbeta. AB - SHP (short heterodimer partner) is an unusual orphan nuclear receptor that contains a putative ligand-binding domain but lacks a conserved DNA-binding domain. Although no conventional receptor function has yet been identified, SHP has been proposed to act as a negative regulator of nuclear receptor signaling pathways, because it interacts with and inhibits DNA binding and transcriptional activity of various nonsteroid receptors, including thyroid hormone and retinoid receptors. We show here that SHP interacts directly with agonist-bound estrogen receptors, ERalpha and ERbeta, and inhibits ER-mediated transcriptional activation. SHP specifically targets the ligand-regulated activation domain AF-2 and competes for binding of coactivators such as TIF2. Thus, SHP may represent a new category of negative coregulators for ligand-activated nuclear receptors. SHP mRNA is widely expressed in rat tissues including certain estrogen target tissues, and subcellular localization studies demonstrate that SHP is a nuclear protein, suggesting a biological significance of the SHP interactions with ERs. Taken together, these results identify ERs as novel SHP targets and suggest that competition for coactivator-binding is a novel mechanism by which SHP may inhibit nuclear receptor activation. PMID- 9867850 TI - In vivo characterization of chimeric phytochromes in yeast. AB - Phytochromes are plant photoreceptors that play a major role in photomorphogenesis. Two members of the phytochrome family have been characterized in some detail. Phytochrome A, which controls very low fluence and high irradiance responses, is rapidly degraded in the light, forms sequestered areas of phytochrome (SAPs), and does not exhibit dark reversion in monocotyledonous seedlings. Phytochrome B mediates red/far-red reversible responses, is stable in the light, and does not form SAPs. We report on the behavior in yeast of the phytochrome apoproteins of rice PHYA, tobacco PHYB, and chimeric PHYAB and PHYBA and on the behavior of the respective holoprotein adducts after assembly with phycocyanobilin chromophore (PHY*). SAP-like formation in yeast was not observed for PHYB, but was detectable for PHYA, PHYAB, and PHYBA. Rice PHYA* did not undergo dark reversion in yeast. Surprisingly, all other tested phytochrome constructs did exhibit dark reversion, including chimeric phytochromes with a short N-terminal part of tobacco PHYB or parsley PHYA fused to rice PHYA. Furthermore, the proportion of phytochrome undergoing dark reversion and the rate of reversion were increased for both the N terminus-swapped constructs and PHYBA*. These results are discussed with respect to structure/function analysis of phytochromes A and B. PMID- 9867852 TI - Functional beta1-integrins release the suppression of fibronectin matrix assembly by vitronectin. AB - beta1-null GD25 fibroblasts adherent to vitronectin fail to bind the N-terminal 70-kDa matrix assembly domain of fibronectin or to assemble fibronectin (Sakai, T., Zhang, Q., Fassler, R., and Mosher, D. F. (1998) J. Cell Biol. 141, 527-538). We have made four observations that extend this finding. First, the presence of vitronectin on a substrate that otherwise can support fibronectin assembly has a dominant-negative effect on assembly. Second, the dominant-negative effect is lost when active beta1A is expressed. Third, beta1A containing the extracellular D130A inactivating mutation has a dominant-negative effect on fibronectin assembly. Fourth, beta1-null cells adherent to vitronectin are flat and lack filopodia, whereas beta1-null cells adherent to fibronectin or beta1A-expressing cells adherent to either vitronectin or fibronectin are contracted and exhibit numerous filopodia. These results reveal, therefore, that GD25 cells adherent to vitronectin can only assume a shape suitable for assembly of fibronectin when there is a countervailing signal from functional beta1-integrins. PMID- 9867851 TI - Intracellular glycerol levels modulate the activity of Sln1p, a Saccharomyces cerevisiae two-component regulator. AB - The HOG mitogen-activated protein kinase pathway mediates the osmotic stress response in Saccharomyces cerevisiae, activating genes like GPD1 (glycerol phosphate dehydrogenase), required for survival under hyperosmotic conditions. Activity of this pathway is regulated by Sln1p, a homolog of the "two-component" histidine kinase family of signal transduction molecules prominent in bacteria. Sln1p also regulates the activity of a Hog1p-independent pathway whose transcriptional output can be monitored using an Mcm1p-dependent lacZ reporter gene. The relationship between the two Sln1p branches is unclear, however, the requirement for unphosphorylated pathway intermediates in Hog1p pathway activation and for phosphorylated intermediates in the activation of the Mcm1p reporter suggests that the two Sln1p branches are reciprocally regulated. To further investigate the signals and molecules involved in modulating Sln1p activity, we have screened for new mutations that elevate the activity of the Mcm1p-dependent lacZ reporter gene. We find that loss of function mutations in FPS1, a gene encoding the major glycerol transporter in yeast activates the reporter in a SLN1-dependent fashion. We propose that elevated intracellular glycerol levels in the fps1 mutant shift Sln1p to the phosphorylated state and trigger the Sln1-dependent activity of the Mcm1 reporter. These observations are consistent with a model in which Sln1p autophosphorylation is triggered by a hypo osmotic stimulus and indicate that the Sln1p osmosensor is tied generally to osmotic balance, and may not specifically sense an external osmolyte. PMID- 9867853 TI - Lipopolysaccharide induction of tissue factor in THP-1 cells involves Jun protein phosphorylation and nuclear factor kappaB nuclear translocation. AB - Tissue Factor (TF) gene expression is transiently induced in human monocytic THP 1 cells by lipopolysaccharide (LPS). We characterized the transcription factor complexes binding to the TF gene promoter LPS response element (LRE) (-220 to 172), which contains binding sites for nuclear factor kappaB (NFkappaB) and activator protein 1 (AP1) transcription factors, and examined the nature of the activation of these factors during a 24-h time course of LPS stimulation. We found proteolysis of the cytoplasmic inhibitory protein IkappaBalpha and nuclear translocation of the NFkappaB/Rel family proteins p65 and c-Rel, corresponding to the transient binding of a p65/c-Rel heterodimer to the kappaB-like site of the LRE. AP1 binding to the LRE was found to be constitutive, with the majority of the AP1 complexes being JunD/Fra-2 heterodimers. A change in the activation state of the AP1 complexes was, however, found to be transient, as determined by JunD phosphorylation of AP1 bound to the proximal binding site. This directly correlates to the transient activation of Jun N-terminal kinase (SAPK/JNK). These data indicate that LPS induction of TF gene expression in monocytic THP-1 cells is regulated by both the transient phosphorylation of Jun-family proteins and the nuclear translocation and transient binding of NFkappaB/Rel proteins. PMID- 9867854 TI - Low density lipoprotein phosphorylates the focal adhesion-associated kinase p125(FAK) in human platelets independent of integrin alphaIIb beta3. AB - Low density lipoprotein (LDL) is known to sensitize platelets to agonists via integrin mediated outside-in signaling (Hackeng, C. M., Huigsloot, M., Pladet, M. W., Nieuwenhuis, H. K., Rijn, H. J. M. v., and Akkerman, J. W. N. (1999) Arterioscler. Thromb. Vasc. Biol., in press). As outside in signaling is associated with phosphorylation of p125(FAK), the effect of LDL on p125(FAK) phosphorylation in platelets was investigated. LDL induced p125(FAK) phosphorylation in a dose- and time- dependent manner. The phosphorylation was independent of ligand binding to integrin alphaIIbbeta3 and aggregation, such in contrast to alpha-thrombin-induced p125(FAK) phosphorylation, that critically depended on platelet aggregation. Platelets from patients with Glanzmann's thrombastenia showed the same LDL- induced phos- phorylation of p125(FAK) as control platelets, whereas alpha-thrombin completely failed to phosphorylate the kinase in the patients platelets. LDL signaling to p125(FAK) was independent of integrin alpha2 beta1, the FcgammaRII receptor, and the lysophosphatidic acid receptor and not affected by inhibitors of cyclooxygenase, protein kinase C, ERK1/2 or p38(MAPK). Phosphorylation of p125(FAK) by LDL was strongly inhibited by cyclic AMP. These observations indicate that LDL is a unique platelet agonist, as it phosphorylates p125(FAK) in platelet suspensions, under unstirred conditions and independent of integrin alphaIIb beta3. PMID- 9867855 TI - Dorsal root ganglia neuron-specific promoter activity of the rabbit beta galactoside alpha1,2-fucosyltransferase gene. AB - The rabbit H-blood type alpha1,2-fucosyltransferase (RFT-I), gene and its biosynthetic products, H antigens (Fucalpha1,2Galbeta), are abundantly expressed in a subset of dorsal root ganglia (DRG) neurons. To investigate the regulatory mechanisms for the RFT-I gene expression, we determined the genomic structure and promoter activity of this gene. PCR amplification of the 5' cDNA end analysis revealed two transcriptional start sites, 498 and 82 nucleotides upstream of the translational initiation codon, the latter site yielding a major 3.1-kb transcript specifically expressed in DRG, as revealed by Northern blotting. Promoter analysis of the 5'-flanking region of the RFT-I gene using a luciferase gene reporter system demonstrated strong promoter activity in PC12 cells, which express the rat H-type alpha1,2-fucosyltransferase gene, and Neuro2a mouse neuroblastoma cells. Deletion analysis revealed the 704-base pair minimal promoter region flanking the translational initiation codon, for which two distinct promoter activities were detected and differentially used in PC12 and Neuro2a cells. The minimal promoter region contained a GC-rich domain (GC content 80%), in which a Sp1 binding sequence and a GSG-like nerve growth factor responsive element were found, but lacked TATA- and CAAT-boxes. Promoter analysis with a primary culture of DRG neurons demonstrated that the minimal promoter region of the RFT-I gene was sufficient for the expression of a reporter gene in DRG neurons. We conclude that the TATA-less GC-rich minimal promoter region of the RFT-I gene controls DRG small neuron-specific expression of the RFT-I gene. PMID- 9867856 TI - Regulation of serine biosynthesis in Arabidopsis. Crucial role of plastidic 3 phosphoglycerate dehydrogenase in non-photosynthetic tissues. AB - In plants, Ser is synthesized through a couple of pathways. 3-Phosphoglycerate dehydrogenase (PGDH), the first enzyme that is involved in the phosphorylated pathway of Ser biosynthesis, is responsible for the oxidation of 3 phosphoglycerate to phosphohydroxypyruvate. Here we report the first molecular cloning and characterization of PGDH from Arabidopsis thaliana. Sequence analysis of cDNA and a genomic clone revealed that the PGDH gene is composed of three exons, encoding a 623-amino acid polypeptide (66, 453 Da). The deduced protein, containing three of the most conserved regions in the NAD-dependent 2-hydroxyacid dehydrogenase family, has 38-39% identity to its animal and bacterial counterparts. The presence of an N-terminal signal sequence for translocation into plastids was confirmed by particle-gun bombardment experiments using green fluorescence protein as a reporter protein for subcellular localization. Southern hybridization analysis and restriction fragment length polymorphism mapping indicated that PGDH is a single-copy gene that is mapped to the upper arm of chromosome 1. Northern hybridization analysis indicated preferential expression of PGDH mRNA in root tissues of light-grown plants, suggesting that the phosphorylated pathway of Ser biosynthesis plays an important role in supplying Ser to non-photosynthetic tissues. The recombinant enzyme overproduced in Escherichia coli displayed hyperbolic kinetics with respect to 3-phosphoglycerate and NAD+. PMID- 9867857 TI - The carboxyl terminus of interferon-gamma contains a functional polybasic nuclear localization sequence. AB - Cytokines such as interferon-gamma (IFN-gamma), which utilize the well studied JAK/STAT pathway for nuclear signal transduction, are themselves translocated to the nucleus. The exact mechanism for the nuclear import of IFN-gamma or the functional role of the nuclear translocation of ligand in signal transduction is unknown. We show in this study that nuclear localization of IFN-gamma is driven by a simple polybasic nuclear localization sequence (NLS) in its COOH terminus, as verified by its ability to specify nuclear import of a heterologous protein allophycocyanin (APC) in standard import assays in digitonin-permeabilized cells. Similar to other nuclear import signals, we show that a peptide representing amino acids 95-132 of IFN-gamma (IFN-gamma(95-132)) containing the polybasic sequence 126RKRKRSR132 was capable of specifying nuclear uptake of the autofluorescent protein, APC, in an energy-dependent fashion that required both ATP and GTP. Nuclear import was abolished when the above polybasic sequence was deleted. Moreover, deletions immediately NH2-terminal of this sequence did not affect the nuclear import. Thus, the sequence 126RKRKRSR132 is necessary and sufficient for nuclear localization. Furthermore, nuclear import was strongly blocked by competition with the cognate peptide IFN-gamma(95-132) but not the peptide IFN-gamma(95-125), which is deleted in the polybasic sequence, further confirming that the NLS properties were contained in this sequence. A peptide containing the prototypical polybasic NLS sequence of the SV40 large T-antigen was also able to inhibit the nuclear import mediated by IFN-gamma(95-132). This observation suggests that the NLS in IFN-gamma may function through the components of the Ran/importin pathway utilized by the SV40 T-NLS. Finally, we show that intact IFN-gamma, when coupled to APC, was also able to mediate its nuclear import. Again, nuclear import was blocked by the peptide IFN-gamma(95 132) and the SV40 T-NLS peptide, suggesting that intact IFN-gamma was also transported into the nucleus through the Ran/importin pathway. Previous studies have suggested a direct intracellular role for IFN-gamma in the induction of its biological activities. Based on our data in this study, we suggest that a key intracellular site of interaction of IFN-gamma is the one with the nuclear transport mechanism that occurs via the NLS in the COOH terminus of IFN-gamma. PMID- 9867858 TI - Phosphorylation of myristoylated alanine-rich protein kinase C substrate by mitogen-activated protein kinase in cultured rat hippocampal neurons following stimulation of glutamate receptors. AB - Glutamate-induced phosphorylation of myristoylated alanine-rich protein kinase C substrate (MARCKS) was investigated in cultured rat hippocampal neurons. In 32P labeled hippocampal neurons, exposure to 10 microM glutamate induced a long lasting increase in phosphorylation of MARCKS. The long lasting increase in MARCKS phosphorylation mainly required activation of the N-methyl-D-aspartate receptor. Unexpectatively, the MARCKS phosphorylation after the 10-min incubation with glutamate was not inhibited by treatment with calphostin C, a potent inhibitor for protein kinase C (PKC), or down-regulation of PKC but was largely prevented by PD098059, a selective inhibitor for mitogen-activated protein (MAP) kinase kinase. In contrast, the phosphorylation following the short exposure to glutamate was prevented by a combination of PD098059 and calphostin C. The phosphopeptide mapping and immunoblotting analyses confirmed that PKC-dependent phosphorylation of MARCKS was transient and the MAP kinase-dependent phosphorylation was relatively persistent. Investigations of the functional properties also showed that the MARCKS phosphorylation by MAP kinase regulates its calmodulin-binding ability and its interaction with F-actin as seen in the PKC-dependent phosphorylation. These results suggest that glutamate causes a long lasting increase in MARCKS phosphorylation through activation of the N-methyl-D aspartate receptor and subsequent activation of MAP kinase in the hippocampal neurons. PMID- 9867859 TI - Incorporation of an isoleucine zipper motif enhances the biological activity of soluble CD40L (CD154). AB - Recent progress in the understanding of immune function indicates that the interaction of CD40L with its receptor, CD40, plays a pivotal role in both humoral immunity and cell-mediated defense against pathogens. Functional studies of this interaction on both dendritic cells and malignant cells have demonstrated that CD40L also plays an important role in immune surveillance and anti-tumor immunity. CD40L exists in nature predominantly as a membrane-anchored molecule. To develop CD40L as a potential therapeutic, it is important to optimize soluble forms of this molecule that could be used in a clinical setting. Several reports have shown that soluble forms of CD40L, like CD40 antibodies, are biologically active. In the present report we demonstrate that the incorporation of an isoleucine zipper trimerization motif significantly enhances the biological activity of soluble CD40L. PMID- 9867860 TI - Saccharomyces cerevisiae GNA1, an essential gene encoding a novel acetyltransferase involved in UDP-N-acetylglucosamine synthesis. AB - The Saccharomyces cerevisiae gene, YFL017C, for a putative acetyltransferase was characterized. Disruption of YFL017C was lethal, leading to a morphology similar to those caused by the depletion of AGM1 or UAP1, the genes encoding phospho-N acetylglucosamine mutase and UDP-N-acetylglucosamine pyrophosphorylase, respectively. This implies the involvement of YFL017C in UDP-N-acetylglucosamine synthesis. The recombinant protein for YFL017C displayed phosphoglucosamine acetyltransferase activities in vitro and utilized glucosamine 6-phosphate as the substrate. When incubated with Agm1p and Uap1p, the Yfl017c protein produced UDP N-acetylglucosamine from glucosamine 6-phosphate. These results indicate that YFL017C specifies glucosamine-6-phosphate acetyltransferase; therefore, the gene was designated GNA1 (glucosamine-6-phosphate acetyltransferase). In addition, whereas bacterial phosphoglucosamine acetyltransferase and UDP-N acetylglucosamine pyrophosphorylase activities are intrinsic in a single polypeptide, they are encoded by distinct essential genes in yeast. When the sequence of ScGna1p was compared with those of other acetyltransferases, Ile97, Glu98, Val102, Gly112, Leu115, Ile116, Phe142, Tyr143, and Gly147 were found to be highly conserved. When alanine was substituted for these amino acids, the enzyme activity for the substituted Phe142 or Tyr143 enzymes was severely diminished. Although the activity of Y143A was too low to perform kinetics, F142A displayed a significantly increased Km value for acetyl-CoA, suggesting that the Phe142 and Tyr143 residues are essential for the catalysis. PMID- 9867861 TI - Interaction of recombinant procollagen and properdin modules of thrombospondin-1 with heparin and fibrinogen/fibrin. AB - Many properties have been assigned to the procollagen and properdin (Type I) modules of thrombospondin-1 (TSP1) based on activities of large proteolytic fragments of TSP1 or peptides containing TSP1-derived sequences. To examine the activities of the modules more exactly, we expressed the first properdin module (P1); the third properdin module (P3); the first and second properdin modules (P12); the first, second, and third properdin modules (P123); and the procollagen module with the first, second, and third properdin modules (CP123) in the GELEX expression vector (GE1) using the baculovirus system. GE1 encodes the pre-pro sequence, the transglutaminase cross-linking site(s), the protease-sensitive site, and the gelatin binding domain from the amino terminus of rat fibronectin. All five recombinant proteins were expressed by insect cells, secreted into the culture medium, and purified by gelatin-agarose affinity chromatography. P123 shared with TSP1 a resistance to trypsin unless reduced and alkylated. P12/GE1, P123/GE1, and CP123/GE1 bound poorly to heparin-agarose except in the absence of sodium chloride, whereas peptides based on P2 are known to bind to heparin in up to 150 mM sodium chloride. In cross-linking experiments employing activated recombinant factor XIII and the transglutaminase cross-linking site in the fibronectin-derived sequence, P12/GE1, P123/GE1, CP123/GE1, and P3/GE1 but not P1/GE1 became incorporated into a fibrin clot more than GE1 alone. Analysis of the complex indicated that cross-linking was to the portion of the fibrin alpha chain remaining in the D-dimer of plasmin digests. P123 also cross-linked to the Aalpha-chain of unclotted fibrinogen. P123 competed for 125I-TSP1 incorporation into the fibrin clot. P123 did not cross-link to plasminogen, histidine-rich glycoprotein, fibronectin, or plasma globulins other than fibrinogen/fibrin. These results indicate that the properdin modules of TSP1 specifically interact with fibrinogen/fibrin but not with heparin under physiologic conditions. PMID- 9867862 TI - PU.1 regulates the CXCR1 promoter. AB - The interleukin-8 receptors (CXCR1 and CXCR2) are specifically expressed at high levels in cells of the neutrophil lineage. In this work we identify promoter elements of the CXCR1 gene and the ets family transcription factor PU.1 as a major regulator for activation of the CXCR1 promoter. We first showed that the upstream sequence of CXCR1 (-800 to +86 base pairs (bp)) directs myeloid-specific expression of reporter gene constructs. Second, we showed the presence of negative elements in the sequence from -800 to -128 bp and positive elements from -128 to +50 bp. Third, we demonstrated that the fragment -22 to +14 bp binds PU.1. Fourth, we showed that PU.1 transactivates the CXCR1 promoter. These data are the first demonstration of PU.1-mediated transcriptional regulation of a neutrophil chemoattractant G protein-coupled receptor. PMID- 9867863 TI - The golden hamster aphrodisin gene. Structure, expression in parotid glands of female animals, and comparison with a similar murine gene. AB - The so-called lipocalins are a family of extracellular proteins that are known to typically fulfill tasks as transport proteins for small hydrophobic molecules. However, in the last decade, a large diversity has been described concerning their functions, for example as enzymes, immunomodulators, or proteins involved in coloration and pheromone action. Aphrodisin belongs to those lipocalins, which are of significant importance for the pheromonal stimulation of copulatory behavior in male hamsters. We recently succeeded in characterizing the corresponding cDNA and demonstrated the expression of the aphrodisin gene in the vagina, uterus, and Bartholin's glands of female hamsters. Here we report the structure of the aphrodisin gene and the functionality of its promoter region. We further compare the aphrodisin gene to the related gene for mouse odorant-binding protein 1a, indicating similar functions of their products. As a novelty, we show that the aphrodisin gene, in addition to the above-mentioned tissues, is also expressed in female hamster parotid glands. In contradiction to the results expected, we finally demonstrate that aphrodisin already occurs in vaginal discharge before the female animals reach fertility. These findings may lead to the identification of as yet unknown aphrodisin functions. PMID- 9867864 TI - Oligomerization and topology of the Golgi membrane protein glucosylceramide synthase. AB - Glucosylceramide synthase (GCS) catalyzes the transfer of glucose from UDP glucose to ceramide to form glucosylceramide, the precursor of most higher order glycosphingolipids. Recently, we characterized GCS activity in highly enriched fractions from rat liver Golgi membranes (Paul, P., Kamisaka, Y., Marks, D. L., and Pagano, R. E. (1996) J. Biol. Chem. 271, 2287-2293), and human GCS was cloned by others (Ichikawa, S., Sakiyama, H., Suzuki, G., Hidari, K. I.-P. J., and Hirabayashi, Y. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 4638-4643). However, the polypeptide responsible for GCS activity has never been identified or characterized. In this study, we made polyclonal antibodies against peptides based on the predicted amino acid sequence of human GCS and used these antibodies to characterize the GCS polypeptide in rat liver Golgi membranes. Western blotting of rat liver Golgi membranes, human cells, and recombinant rat GCS expressed in bacteria showed that GCS migrates as an approximately 38-kDa protein on SDS-polyacrylamide gels. Trypsinization and immunoprecipitation studies with Golgi membranes showed that both the C terminus and a hydrophilic loop near the N terminus of GCS are accessible from the cytosolic face of the Golgi membrane. Treatment of Golgi membranes with N-hydroxysuccinimide ester-based cross-linking reagents yielded an approximately 50-kDa polypeptide recognized by anti-GCS antibodies; however, treatment of approximately 10,000-fold purified Golgi GCS with the same reagents did not yield cross-linked GCS forms. These results suggest that GCS forms a dimer or oligomer with another protein in the Golgi membrane. The migration of solubilized Golgi GCS in glycerol gradients was also consistent with a predominantly oligomeric organization of GCS. PMID- 9867865 TI - Topology of Euglena chloroplast protein precursors within endoplasmic reticulum to Golgi to chloroplast transport vesicles. AB - Euglena chloroplast protein precursors are transported as integral membrane proteins from the endoplasmic reticulum (ER) to the Golgi apparatus prior to chloroplast localization. All Euglena chloroplast protein precursors have functionally similar bipartite presequences composed of an N-terminal signal peptide domain and a stromal targeting domain containing a hydrophobic region approximately 60 amino acids from the predicted signal peptidase cleavage site. Asparagine-linked glycosylation reporters and presequence deletion constructs of the precursor to the Euglena light-harvesting chlorophyll a/b-binding protein of photosystem II (pLHCPII) were used to identify presequence regions translocated into the ER lumen and stop transfer membrane anchor domains. An asparagine-linked glycosylation site present at amino acid 148 of pLHCPII near the N terminus of mature LHCPII was not glycosylated in vitro by canine microsomes while an asparagine-linked glycosylation site inserted at amino acid 40 was. The asparagine at amino acid 148 was glycosylated upon deletion of amino acids 46 146, which contain the stromal targeting domain, indicating that the hydrophobic region within this domain functions as a stop transfer membrane anchor sequence. Protease protection assays indicated that for all constructs, mature LHCPII was not translocated across the microsomal membrane. Taken together with the structural similarity of all Euglena presequences, these results demonstrate that chloroplast precursors are anchored within ER and Golgi transport vesicles by the stromal targeting domain hydrophobic region oriented with the presequence N terminus formed by signal peptidase cleavage in the vesicle lumen and the mature protein in the cytoplasm. PMID- 9867866 TI - IQGAP1 integrates Ca2+/calmodulin and Cdc42 signaling. AB - Calmodulin regulates diverse Ca2+-dependent cellular processes, including cell cycle progression and cytoskeletal rearrangement. A recently identified calmodulin-binding protein, IQGAP1, interacts with both actin and Cdc42. In this study, evidence is presented that, in the absence of Ca2+, IQGAP1 bound to Cdc42, which maintained Cdc42 in the active GTP-bound state. Addition of Ca2+ both directly abrogated the effect of IQGAP1 on the intrinsic GTPase activity of Cdc42 and, in the presence of calmodulin, dissociated Cdc42 from IQGAP1. In addition, in vitro binding assays revealed that calmodulin associated with both the calponin homology domain and the IQ motifs of IQGAP1. Moreover, F-actin competed with Ca2+/calmodulin for binding to the calponin homology domain, but not the IQ motifs, of IQGAP1. Analysis of cell lysates revealed that calmodulin bound to IQGAP1 in a ternary complex with Cdc42. Increasing the Ca2+ concentration enhanced the interaction between calmodulin and IQGAP1, with a concomitant decrease in the association of IQGAP1 with Cdc42. Our data suggest that IQGAP1 functions as a scaffolding protein, providing a molecular link between Ca2+/calmodulin and Cdc42 signaling. PMID- 9867867 TI - Cloning, expression, and nutritional regulation of the mammalian Delta-6 desaturase. AB - Arachidonic acid (20:4(n-6)) and docosahexaenoic acid (22:6(n-3)) have a variety of physiological functions that include being the major component of membrane phospholipid in brain and retina, substrates for eicosanoid production, and regulators of nuclear transcription factors. The rate-limiting step in the production of 20:4(n-6) and 22:6(n-3) is the desaturation of 18:2(n-6) and 18:3(n 3) by Delta-6 desaturase. In this report, we describe the cloning, characterization, and expression of a mammalian Delta-6 desaturase. The open reading frames for mouse and human Delta-6 desaturase each encode a 444-amino acid peptide, and the two peptides share an 87% amino acid homology. The amino acid sequence predicts that the peptide contains two membrane-spanning domains as well as a cytochrome b5-like domain that is characteristic of nonmammalian Delta 6 desaturases. Expression of the open reading frame in rat hepatocytes and Chinese hamster ovary cells instilled in these cells the ability to convert 18:2(n-6) and 18:3(n-3) to their respective products, 18:3(n-6) and 18:4(n-3). When mice were fed a diet containing 10% fat, hepatic enzymatic activity and mRNA abundance for hepatic Delta-6 desaturase in mice fed corn oil were 70 and 50% lower than in mice fed triolein. Finally, Northern analysis revealed that the brain contained an amount of Delta-6 desaturase mRNA that was several times greater than that found in other tissues including the liver, lung, heart, and skeletal muscle. The RNA abundance data indicate that prior conclusions regarding the low level of Delta-6 desaturase expression in nonhepatic tissues may need to be reevaluated. PMID- 9867868 TI - Nuclear extracts lacking DNA-dependent protein kinase are deficient in multiple round transcription. AB - We have compared levels of in vitro transcription in nuclear extracts from DNA dependent protein kinase (DNA-PK)-deficient and DNA-PK-containing Chinese hamster ovary cell lines. DNA-PK-deficient cell lines are radiosensitive mutants lacking either the catalytic subunit or the 80-kDa subunit of the Ku protein regulatory component. Extracts from DNA-PK-deficient cell lines had a 2-7-fold decrease in the level of in vitro transcription when compared with matched controls. This decrease was observed with several promoters. Transcription could be restored to either of the deficient extracts by addition of small amounts of extract from the DNA-PK-containing cell lines. Transcription was not restored by addition of purified DNA-PK catalytic subunit, Ku protein, or individually purified general transcription factors. We conclude that extracts from DNA-PK-deficient cells lack a positively acting regulatory factor or a complex of factors not readily reconstituted with individual proteins. We have also investigated the mechanistic defect in the deficient extracts and have found that the observed differences in transcription levels between Ku-positive and Ku-negative cell lines can be attributed solely to a greater ability of the Ku-positive nuclear extracts to carry out secondary initiation events subsequent to the first round of transcription. PMID- 9867869 TI - The dynamic role of GRP78/BiP in the coordination of mRNA translation with protein processing. AB - The role of GRP78/BiP in coordinating endoplasmic reticular (ER) protein processing with mRNA translation was examined in GH3 pituitary cells. ADP ribosylation of GRP78 and eukaryotic initiation factor (eIF)-2alpha phosphorylation were assessed, respectively, as indices of chaperone inactivation and the inhibition of translational initiation. Inhibition of protein processing by ER stress (ionomycin and dithiothreitol) resulted in GRP78 deribosylation and eIF-2 phosphorylation. Suppression of translation relative to ER protein processing (cycloheximide) produced approximately 50% ADP-ribosylation of GRP78 within 90 min without eIF-2 phosphorylation. ADP-ribosylation was reversed in 90 min by cycloheximide removal in a manner accelerated by ER stressors. Cycloheximide sharply reduced eIF-2 phosphorylation in response to ER stressors for about 30 min; sensitivity returned as GRP78 became increasingly ADP ribosylated. Reduced sensitivity of eIF-2 to phosphorylation appeared to derive from the accumulation of free, unmodified chaperone as proteins completed processing without replacements. Prolonged (24 h) incubations with cycloheximide resulted in the selective loss of the ADP-ribosylated form of GRP78 and increased sensitivity of eIF-2 phosphorylation in response to ER stressors. Brefeldin A decreased ADP-ribosylation of GRP78 in parallel with increased eIF-2 phosphorylation. The cytoplasmic stressor, arsenite, which inhibits translational initiation through eIF-2 phosphorylation without affecting the ER, also produced ADP-ribosylation of GRP78. PMID- 9867870 TI - Molecular analysis of mammalian phospholipase D2. AB - The mammalian phosphatidylcholine-specific phospholipase D (PLD) enzymes PLD1 and PLD2 have been proposed to play roles in signal transduction and membrane vesicular trafficking in distinct subcellular compartments. PLD1 is activated in a synergistic manner in vitro by protein kinase C-alpha, ADP-ribosylation factor 1 (ARF1), and Rho family members. In contrast, PLD2 is constitutively active in vitro. We describe here molecular analysis of PLD2. We show that the NH2-terminal 308 amino acids are required for PLD2's characteristic high basal activity. Unexpectedly, PLD2 lacking this region becomes highly responsive to ARF proteins and displays a modest preference for activation by ARF5. Chimeric analysis of PLD1 and PLD2 suggests that the ARF-responsive region is in the PLD carboxyl terminus. We also inserted into PLD2 a region of sequence unique to PLD1 known as the "loop" region, which had been proposed initially to mediate effector stimulation but that subsequently was shown instead to be required in part for the very low basal activity characteristic of PLD1. The insertion decreased PLD2 activity, consistent with the latter finding. Finally, we show that the critical role undertaken by the conserved carboxyl terminus is unlikely to involve promoting PLD association with membrane surfaces. PMID- 9867871 TI - The human copper-zinc superoxide dismutase gene (SOD1) proximal promoter is regulated by Sp1, Egr-1, and WT1 via non-canonical binding sites. AB - Human copper-zinc superoxide dismutase (Cu,Zn-SOD) participates in the control of reactive oxygen intermediate intracellular concentration. In this study, we show that phorbol 12-myristate 13-acetate (PMA) increases Cu,Zn-SOD mRNA expression within 30 min. The sequence between nucleotides -71 and -29 is essential for both basal and PMA-induced gene expression. This region includes an Sp1-binding site that is also recognized by a possible Sp1-like protein and by Egr-1 in a PMA inducible manner. Egr-1 and two splicing variants of the Egr-related protein WT1 were able to transactivate the SOD1 promoter in co-transfection experiments. Sp1 and the possible Sp1-like proteins bind to two overlapping, but distinct sequences. However, Egr-1 and Sp1 seem to interact with two sites that are either identical or very close to each other. None of these sites fit the consensus sequences previously reported for these proteins. Analysis of various mutants of the SOD1 proximal promoter revealed that the region that binds Sp1 and Egr-1 is required for both basal and Egr-1-driven expression. Interplay between different members of the Sp1 family, Egr-1, and different splicing variants of WT1 in the SOD1 proximal promoter may provide clues about the physiological function of Cu,Zn-SOD. PMID- 9867872 TI - Ah receptor and NF-kappaB interactions, a potential mechanism for dioxin toxicity. AB - The Ah receptor (AhR) mediates many of the toxic responses induced by polyhalogenated and polycyclic hydrocarbons (PAHs) which are ubiquitous environmental contaminants causing toxic responses in human and wildlife. NF kappaB is a pleiotropic transcription factor controlling many physiological functions adversely affected by PAHs, including immune suppression, thymus involution, hyperkeratosis, and carcinogenesis. Here, we show physical interaction and mutual functional repression between AhR and NF-kappaB. This mutual repression may provide an underlying mechanism for many hitherto poorly understood PAH-induced toxic responses, and may also provide a mechanistic explanation for alteration of xenobiotic metabolism by cytokines and compounds that regulate NF-kappaB. PMID- 9867873 TI - Thioredoxin activation of iron regulatory proteins. Redox regulation of RNA binding after exposure to nitric oxide. AB - Iron regulatory proteins (IRP1 and IRP2) are redox-sensitive RNA-binding proteins that modulate the expression of several genes encoding key proteins of iron metabolism. IRP1 can also exist as an aconitase containing a [4Fe-4S] cluster bound to three cysteines at the active site. We previously showed that biosynthesis of nitric oxide (NO) induces the transition of IRP1 from aconitase to apoprotein able to bind RNA. This switch is also observed when cytosolic extracts are exposed to NO donors. However, the activation of IRP1 under these conditions is far from maximal. In this study we examined the capacity of physiological reducing systems to cooperate with NO in the activation of IRP1. Cytosolic extracts from the macrophage cell line RAW 264.7 or purified IRP1 were incubated with NO donors and subsequently exposed to glutathione or to thioredoxin (Trx), a strong protein disulfide reductase. Trx was the most effective, inducing a 2-6-fold enhancement of the RNA binding activity of NO treated IRP1. Furthermore, the effect of NO on IRP1 from cytosolic extracts was abolished in the presence of anti-Trx antibodies. We also studied the combined effect of NO and Trx on IRP2, which exhibits constitutive RNA binding activity. We observed an inhibition of IRP2 activity following exposure to NO donors which was restored by Trx. Collectively, these results point to a crucial role of Trx as a modulator of IRP activity in situations of NO production. PMID- 9867874 TI - Proinflammatory macrophage-activating properties of the novel phospholipid diacylglycerol pyrophosphate. AB - We have found that the novel phospholipid diacylglycerol pyrophosphate (DGPP), identified in bacteria, yeast, and plants, but not in mammalian cells, is able to potently activate macrophages for enhanced secretion of arachidonate metabolites, a key event in the immunoinflammatory response of leukocytes. Macrophage responses to DGPP are specific and are not mediated by its conversion into other putative lipid mediators such as phosphatidic acid, lysophosphatidic acid, or diacylglycerol. The responses to DGPP are compatible with a receptor-recognition event because they are blocked by suramin. Intracellular signaling initiated by DGPP includes phosphorylation and activation of the Group IV cytosolic phospholipase A2 and of the extracellular-signal regulated p42 mitogen-activated protein kinase (MAPK) and p44 MAPK, and membrane translocation of the protein kinase C isoenzymes alpha, epsilon, delta. These results establish DGPP as a novel macrophage-activating factor and suggest a potential role for this compound in triggering homeostatic cellular responses. PMID- 9867875 TI - Protein kinase C phosphorylates plasma membrane Ca2+ pump isoform 4a at its calmodulin binding domain. AB - Phosphorylation by protein kinase C of isoform 4a of the human plasma membrane Ca2+ pump (hPMCA4a) was studied using the COS cell expression system. Phosphorylation of several truncated mutants of hPMCA4a indicated that a single phosphorylation site lies in a region between residues 1113 and 1125. This region is within the calmodulin binding domain and contains a single phosphorylatable residue, serine 1115. Converting this serine to an alanine diminished phosphorylation greatly. Phosphorylation, done in the absence of calmodulin, did not affect subsequent calmodulin binding, but previous binding of calmodulin did inhibit phosphorylation. Moreover, no significant shift in the calmodulin response curve of hPMCA4a was observed when phosphorylation was mimicked by converting serine 1115 to an acidic residue. The calmodulin binding domain of hPMCA4a is much longer than other calmodulin binding domains and has been suggested to consist of two binding lobes interrupted by a short nonbinding region. The findings of this study indicate that serine 1115 is the residue phosphorylated by protein kinase C, and that it lies within the nonbinding region of the calmodulin binding domain. PMID- 9867876 TI - Requirement of N-terminal cysteines of PSD-95 for PSD-95 multimerization and ternary complex formation, but not for binding to potassium channel Kv1.4. AB - The PSD-95 family of PSD-95/Discs large/ZO-1 (PDZ) domain-containing proteins plays a role in the clustering and localization of specific ion channels and receptors at synapses. Previous studies have shown that PSD-95 forms multimers through an N-terminal region (termed the N-segment) and that the multimerization of PSD-95 is critical for its ability to cluster Shaker-type potassium channel Kv1.4 in heterologous cells. We show here that the PSD-95 N-segment functions as a multimerization domain only when located at the N-terminal end of a heterologous protein. A pair of N-terminal cysteines, Cys3 and Cys5, is essential for the ability of PSD-95 to self-associate and to form cell surface clusters with Kv1.4. However, PSD-95 mutants lacking these cysteine residues retain their ability to associate with membranes and to bind to Kv1.4. Unlike wild type PSD 95, the cysteine mutant of PSD-95 cannot form a ternary complex with Kv1.4 and the cell adhesion molecule Fasciclin II. These results suggest that the N terminal cysteines are essential for PSD-95 multimerization and that multimerization is required for simultaneous binding of multiple membrane protein ligands by PSD-95. PMID- 9867877 TI - Activation of p38 mitogen-activated protein kinase by lipopolysaccharide in human neutrophils requires nitric oxide-dependent cGMP accumulation. AB - This study examined the signal transduction pathway(s) leading to phosphorylation of p38 in human neutrophils stimulated with lipopolysaccharide and formyl peptides. Blockade of the nitric oxide (NO) pathway in neutrophils with the NO synthase inhibitor N-nitro-L-arginine methyl ester or by treatment with the NO scavenger 2-phenyl-tetramethylimidazoline-1-oxyl-3-oxide attenuated phosphorylation of the mitogen-activated protein kinase p38 in response to lipopolysaccharide but not fMet-Leu-Phe. Using the NO releasing agents S-nitroso N-acetylpenicillamine and sodium nitroprusside it was determined that nitric oxide is sufficient to cause an increase in phosphorylation of p38. Increasing cellular cGMP with phosphodiesterase inhibitors, by stimulation of soluble guanylyl cyclase with YC-1 or with exogenous dibutyryl cGMP resulted in mitogen activated protein kinase/extracellular signal-regulated kinase kinase 3,6 (MEK3,6) activation and phosphorylation of p38. This phenomenon was specific for MEK3,6, because these agents had no effect on the phosphorylation state of MEK1,2. A role for protein kinase G but not protein kinase A downstream of lipopolysaccharide but not formylmethionylleucylphenylalanine was shown using the specific inhibitors KT5823 and H89, respectively. These data indicate that activation of p38 by fMet-Leu-Phe and lipopolysaccharide involve different mechanisms, and that activation of protein kinase G by NO-dependent stimulation of guanylyl cyclase is necessary and sufficient for phosphorylation of p38 downstream of lipopolysaccharide. PMID- 9867879 TI - State of the journal PMID- 9867878 TI - Isolation of supernumerary yeast ATP synthase subunits e and i. Characterization of subunit i and disruption of its structural gene ATP18. AB - Two subunits of the yeast ATP synthase have been isolated. Subunit e was found loosely associated to the complex. Triton X-100 at a 1% concentration removed this subunit from the ATP synthase. The N-terminal sequencing of subunit i has been performed. The data are in agreement with the sequence of the predicted product of a DNA fragment of Saccharomyces cerevisiae chromosome XIII. The ATP18 gene encodes subunit i, which is 59 amino acids long and corresponds to a calculated mass of 6687 Da. Its pI is 9.73. It is an amphiphilic protein having a hydrophobic N-terminal part and a hydrophilic C-terminal part. It is not apparently related to any subunit described in other ATP synthases. The null mutant showed low growth on nonfermentable medium. Mutant mitochondria display a low ADP/O ratio and a decrease with time in proton pumping after ATP addition. Subunit i is associated with the complex; it is not a structural component of the enzyme but rather is involved in the oxidative phosphorylations. Similar amounts of ATP synthase were measured for wild-type and null mutant mitochondria. Because 2-fold less specific ATPase activity was measured for the null mutant than for the wild-type mitochondria, we make the hypothesis that the observed decrease in the turnover of the mutant enzyme could be linked to a proton translocation defect through F0. PMID- 9867880 TI - Acute stroke: delays to presentation and emergency department evaluation. AB - STUDY OBJECTIVE: To document prehospital and inhospital time intervals from stroke onset to emergency department evaluation and to identify factors associated with presentation to the ED within 3 hours of symptom onset, the current time window for thrombolytic therapy. METHODS: Patients admitted through the ED with a diagnosis of stroke were identified through admitting logs. Time intervals were obtained from EMS runsheets and ED records. Information regarding first medical contact, education, and income was obtained by patient interview. Baseline variables were analyzed to assess association with ED arrival within 3 hours of symptom onset; variables significant on univariate analysis were placed in a multivariable model. RESULTS: There were 151 stroke patients (59% white and 41% black). Time of stroke onset and time to ED arrival were documented for 119 patients (79%). The median time from stroke onset to ED arrival was 5.7 hours; 46 patients (30%) presenting within 3 hours. Of those with times recorded, the median time from stroke onset to EMS arrival was 1.7 hours. Multivariable logistic regression identified use of EMS (odds ratio [OR], 4.0; 95% confidence interval [CI], 1.3 to 12.1) and white race (OR, 3.5; 95% CI, 1.3 to 10) as being independently associated with ED arrival within 3 hours of symptom onset. Median time from ED arrival to physician evaluation was 20 minutes. Median time from ED arrival to computed tomographic evaluation was 72 minutes. When patients were asked the main reason they sought medical attention, 40% (60/141) of those able to be interviewed said that they themselves did not decide to seek medical attention, but rather a friend or family member told them they should go to the hospital. CONCLUSION: The median time from stroke onset to ED evaluation was 5.7 hours, with almost a third of patients presenting within 3 hours. Use of EMS and white race were independently associated with arrival within 3 hours. PMID- 9867881 TI - Thrombolytic therapy for acute ischemic stroke: why the majority of patients remain ineligible for treatment. AB - STUDY OBJECTIVES: Thrombolytic therapy has been advocated as an effective treatment for acute ischemic stroke. In an attempt to promote maximum benefit while reducing the risk of hemorrhagic complications, numerous exclusions to the use of thrombolytic therapy for acute ischemic stroke have been promulgated. This study was conducted to identify the number of acute ischemic stroke patients eligible for thrombolytic therapy and to determine the reasons those deemed ineligible were excluded. METHODS: This observational study was conducted from September 15, 1996, to May 1, 1997, at an emergency department with an annual census of 70,000. Patients with a chief complaint suggestive of acute ischemic stroke were categorized as "eligible" if thrombolytic therapy was not contraindicated and could be initiated within 3 hours of symptom onset. Patients were deemed "ineligible" if the time to thrombolytic therapy would have exceeded 3 hours, or if other specific contraindications to thrombolytic therapy were present. For all categories, 95% confidence intervals (95% CI) were determined. RESULTS: Of the 214 patients with acute ischemic stroke who were screened, 6 (2.8%+/-2.2%) were eligible. Ninety-five patients (44%+/-7%) were excluded solely on the basis of the time interval from onset of symptoms to eligibility for thrombolytic therapy exceeding 3 hours. Other common reasons for exclusion included resolution of symptoms in 31 patients (14%+/-4%), intracranial hemorrhage as determined by head computed tomography in 22 (10%+/-4%), and minor symptoms in 20 (9%+/-4%). CONCLUSION: The majority of acute ischemic stroke patients do not meet accepted criteria for thrombolytic therapy. Most are ineligible because of delays in obtaining treatment. Strategies should be devised to reduce the time to treatment if thrombolytic therapy is to achieve widespread use in the treatment of acute ischemic stroke. PMID- 9867882 TI - Use of electron-beam computed tomography in the evaluation of chest pain patients in the emergency department. AB - STUDY OBJECTIVE: We sought to determine whether electron-beam computed tomography (EBCT) could be used as a triage tool in the emergency department for patients with angina-like chest pain, no known history of coronary disease, normal or indeterminate ECG findings, and normal initial cardiac enzyme concentrations. METHODS: We conducted a prospective observational study of 105 patients admitted between December 1995 and October 1997 to the ED of a large tertiary care hospital with 70,000 annual ED visits. The study group was comprised of women aged 40 to 65 years and men aged 30 to 55 years who presented with angina-like chest pain requiring admission to the hospital or chest pain observation unit. All patients underwent EBCT of the coronary arteries, along with other cardiac testing as deemed necessary by staff physicians. RESULTS: Of the 105 patients, 100 underwent other cardiac testing during hospitalization. Evaluation included treadmill exercise testing in 58, coronary angiography in 25, radionuclide stress testing in 19, and echocardiography in 11. Results of EBCT and cardiac testing were negative for both in 53 patients (53%), positive for both in 14 (14%), positive for tomography and negative for cardiac testing in 32 (32%), and negative for tomography and positive for cardiac testing in only 1 patient. This positive test result, on a treadmill exercise test, was ruled a false positive by an independent staff cardiologist. Two other female patients with normal exercise sestamibi or coronary angiography and EBCT findings also had false-positive treadmill exercise results. The sensitivity of EBCT was 100% (95% confidence interval, 77% to 100%), with a negative predictive value of 100% (95% confidence interval, 94% to 100%). Specificity was 63% (95% confidence interval, 54% to 75%). CONCLUSION: EBCT is a rapid and efficient screening tool for patients admitted to the ED with angina-like chest pain, normal cardiac enzyme concentrations, indeterminate ECG findings, and no history of coronary artery disease. Our study suggests that patients with normal initial cardiac enzyme concentrations, normal or indeterminate ECG findings, and negative results on EBCT may be safely discharged from the ED without further testing or observation. Larger studies are required to confirm this conclusion. PMID- 9867883 TI - American Board of Emergency Medicine Longitudinal Study of Emergency Physicians. AB - STUDY OBJECTIVE: The American Board of Emergency Medicine (ABEM) Longitudinal Study of Emergency Physicians (LSEP) was initiated to describe the development of a new medical specialty through the continuing study of the lives of representative emergency physicians. The study is designed to gather data periodically over many years to come. The primary purpose of this article is to provide a baseline for the information obtained and the methods used to develop the ABEM LSEP. METHODS: Stratified, random sampling was used to select emergency physicians who represent different stages in the development of the specialty. Major data collections are conducted using a comprehensive questionnaire in 5 year intervals. Practice profiles were developed and analyzed. Homogeneous scales were created in 9 survey categories and analyzed. RESULTS: The survey was returned by 95% (958/1,008) of the emergency physicians. They are primarily middle-aged, family- and community-oriented, satisfied with their careers, and find that work stress is not a serious problem. Those who are trained in emergency medicine are the most likely to be involved in academic emergency medicine. CONCLUSION: The LSEP is a broad-reaching investigation of emergency physicians. Over time the study will describe (1) the individuals who move the specialty forward at different stages in the growth of the specialty, (2) the realities of practice in the specialty, (3) the relationship of the specialty to the personal lives and well-being of the specialty physicians, and (4) the changes seen in these factors over time. PMID- 9867884 TI - Critical care transport: outcome evaluation after interfacility transfer and hospitalization. AB - STUDY OBJECTIVE: To test the hypothesis that interfacility transfer is not associated with increased mortality, duration of stay, or readmission within 7 days. METHODS: We matched 3,298 patients who were hospitalized for chest pain or related complaints in Kaiser Permanente medical centers after transfer from the emergency department of a nonplan hospital (transported patients) with 3,298 patients of the same gender and age (+/-5 years) and with the same principal diagnosis who were hospitalized within 6 months without transfer in the same Kaiser Permanente medical center (directly admitted patients). Patients were compared in terms of outcome measures: in-hospital deaths, continued care in another facility, readmission within 7 days, in-patient length of stay (LOS), and LOS in special care units. RESULTS: The adjusted odds ratios for in-hospital mortality and readmission within 7 days were 1.0 (95% confidence interval,.8 to 1.4) and.9 (95% confidence interval,.7 to 1.2), respectively. The adjusted mean difference in LOS was -.1 days (95% confidence interval, -.2 to.1). Transported and directly admitted cardiac patients were also compared for all examined outcome measures at each of 10 medical centers. At a few medical centers, we observed significant difference in LOS, special care LOS, and continued care in another facility. However, all these differences were small, and most were probably random errors. CONCLUSION: Conservative patient selection criteria, pretransfer stabilization, and the use of appropriate equipment and medical personnel have resulted in the interfacility transfer program's achieving its goal of transferring high-risk patients without adverse impact on clinical outcomes or resource use. PMID- 9867885 TI - Modifiable factors associated with improved cardiac arrest survival in a multicenter basic life support/defibrillation system: OPALS Study Phase I results. Ontario Prehospital Advanced Life Support. AB - STUDY OBJECTIVES: This study was conducted to identify modifiable factors associated with survival for prehospital cardiac arrest in a large, multicenter EMS system with basic life support/defibrillation (BLS-D) level of care. METHODS: This observational cohort study constitutes Phase I of the 3-phase Ontario Prehospital Advanced Life Support (OPALS) Study. Included were all adults who had cardiac arrest before EMS arrival in 21 urban/suburban communities that operate under the jurisdiction of 1 ambulance services branch, have 911 telephone service, and provide ambulance defibrillation but no prehospital advanced life support (ALS). Central dispatch and ambulance records were reviewed according to the Utstein guidelines. Associations between multiple patient and EMS factors and survival to discharge were assessed by univariate then stepwise logistic regression analyses. RESULTS: From January 1, 1991, to January 31, 1995, 5,335 eligible patients were treated. Of these, 46.8% of cardiac arrests were witnessed by citizens, 14.5% received bystander CPR, 25.6% received CPR by fire or police, and 38.2% had an initial rhythm of ventricular fibrillation/ventricular tachycardia (VF/VT). The mean interval from call received to vehicle stopped was 6.7 minutes. Survival was 3.5% overall and 8.8% for VF/VT. Multivariate analysis found the following factors to be independently associated with survival (odds ratio with 95% confidence intervals): age.81 (. 73,.89), bystander-witnessed arrest 4.05 (2.78, 5.90), bystander CPR 2.98 (2.07, 4.29), CPR by fire or police 2.20 (1.46, 3.31), and response interval call received to vehicle stopped.76 (.71,.82). CONCLUSION: This represents the largest multicenter BLS-D study of prehospital cardiac arrest yet conducted and clearly indicates that patient survival may be improved by optimization of EMS response intervals, bystander CPR, as well as first-responder CPR by fire or police. PMID- 9867886 TI - A new approach to teaching prehospital trauma care to paramedic students. AB - STUDY OBJECTIVE: A modification of the standard Department of Transportation student paramedic curriculum encouraging individualized patient assessment decreases inappropriate on-scene procedures (OSPs) and scene time, measured on simulated patients. METHODS: Scenario-based testing from 1991 through 1993 was videotaped for all students. A new trauma curriculum was introduced in 1992, individualizing patient assessment and prioritization of OSPs. Recorded OSPs included spinal immobilization, application of military antishock trousers, endotracheal intubation, cricothyrotomy, intravenous catheter insertion, and needle thoracostomy. Twenty videotaped random student performances of the 1991 class was compared with a similar sample of 20 from the 1993 class; scene times and the OSP numbers were measured. Two board-certified independent emergency physicians unfamiliar with the students or the new curriculum reviewed all 40 tests on a master videotape. Patient assessment appropriateness, scene time, OSPs, scenario difficulty, and number of inappropriate OSPs were evaluated using a linear analog scale. Data are presented as means with confidence intervals (CIs), analyzed by Student's t test and the Mann-Whitney 2-sample test. RESULTS: Scene time from 1991 to 1993 decreased overall with a mean of 4.3 minutes (95% CI 2.8 to 5.8 minutes), as did the number of OSPs: 3.1 versus 1.7 (mean difference, 1.45 OSPs per scenario; 95% CI.91 to 1.99). Physician reviewers noted improvements in the appropriateness of patient assessment, scene time, and OSPs from 1991 to 1993. There was no significant difference in scenario difficulty for 1991 compared with 1993. Inappropriate OSPs done on scene declined. Physician 1 indicated a mean of inappropriate procedures of 1.6 in 1991 versus.5 in 1993. Physician 2 indicated a mean of 1.4 in 1991 versus.3 in 1993. CONCLUSION: This new paramedic curriculum decreased on-scene time and inappropriate use of procedures in stabilizing the condition of patients with simulated critical trauma. PMID- 9867888 TI - Effects of an emergency department-based advocacy program for battered women on community resource utilization. AB - STUDY OBJECTIVE: The emergency department is often accessed by battered women. Although it is recommended that information about community resources be given to women, there is little information regarding how often women use these resources. The objective of this study was to determine whether an ED-based advocacy program resulted in increased community resource utilization by battered women. METHODS: The study used a before-after trial design at an urban county hospital ED. Before the program, a consecutive sample of 117 women was identified in the ED as having sustained injuries by domestic violence. After the program, a consecutive sample of 105 (57% of the 183 asked) who agreed to meet with an advocate in the ED was identified. Participants in both groups lived in Kansas City, Missouri, and were 18 years of age or older. The outcome measures were the proportion of women with shelter use, shelter-based counseling, police calls, full orders of protection, and repeat ED visits for domestic violence after the index ED visit. RESULTS: After the initiation of the program, shelter use increased from 11% to 28% (P =.003) and shelter-based counseling increased from 1% to 15% (P <.001). There was no change in repeat police calls (25% versus 35%, P =.14), full orders of protection (9% versus 6%, P =. 58), or repeat ED visits for domestic violence (11% versus 8%, P =. 63). CONCLUSION: ED-based advocacy for domestic violence resulted in increased use of shelters and shelter-based counseling. PMID- 9867887 TI - Injuries in a 1-day recreational cycling tour: Bike New York. AB - STUDY OBJECTIVE: To describe injuries during a 1-day urban cycling tour. METHODS: During the May 1996 "Bike New York" tour, we monitored EMS calls to identify injuries in a cohort of helmeted cyclists shielded from traffic. We collected demographic information from entry records, injury data from ambulance call reports, and follow-up on transported patients from telephone interviews with emergency physicians. Data were summarized using proportions, relative risks (RRs), 95% confidence intervals (CIs), and chi2 RESULTS: Approximately 28,000 cyclists participated, of which 23,502 (84%) were officially registered. Sixty eight percent of registered bicyclists were male, and 92% were between 18 and 55 years old. Of the 140 EMS calls made during the tour, 136 (97%) involved participants; this yielded an injury incidence of 5 per 1,000 riders, or 12 to 13 per 100,000 person-miles. Injury was more common among younger cyclists (RR=1.4 for age 35 years; 95% CI, 1.0 to 2.0; P <.05), and possibly women (RR=1.3; 95% CI,.9 to 1.8; P =.11). Injuries were mostly minor, but there were 7 concussions and 6 clavicle fractures; none of the 140 injuries was fatal. Thirty-eight calls resulted in ED transport, and 5 of these patients were admitted. Although EMS units were evenly distributed along the route, most EMS calls occurred in only 3 of the 7 zones (P <.001). CONCLUSION: Injuries during the largest 1-day US cycling tour were uncommon. More data are needed to determine the relative importance of injury risk factors. Data collection during mass events may help guide distribution of EMS personnel. PMID- 9867889 TI - Descriptive epidemiology of injury and illness among cruise ship passengers. AB - STUDY OBJECTIVE: To provide information, which can be used in the formation of guidelines concerning medical facilities and staff on cruise ships, on the descriptive epidemiology of the medical conditions encountered by cruise ship physicians. METHODS: A retrospective descriptive epidemiologic study design was used to evaluate patient physician encounters on cruises originating in a calendar-year period for the 4 ships of a major cruise ship line with cruises originating in the United States. Demographic data regarding sex and age of the passengers on these ships were available for each cruise. We collected information on patient age, sex, chief complaint, diagnoses, treatment, and patient disposition recorded in the patients' medical records in the ships' medical logs. RESULTS: Seven thousand one hundred forty-seven new patient visits occurred in a population of 196,171 passengers and 1,537,298 passenger days; 56.7% of passengers were female, and 60.7% of patients were female; 43.3% of passengers and 39.6% of patients were male. Visits to the ship infirmaries were made for the following reasons: 18.2% of visits were related to injuries, 69.3% were related to medical conditions, and 12.5% were unspecified or other conditions. The most common diagnosis was respiratory tract infection (29.1%); 11% of patients had a serious or potentially life-threatening diagnosis. The most common group of prescription medications prescribed was antibiotics. CONCLUSION: Many different injuries and illnesses occur on board cruise ships. The spectrum is similar in many respects to the patients presenting to emergency departments. Cruise lines must prepare for the initial treatment and stabilization of patients with serious illnesses or injuries with appropriately qualified and equipped medical personnel and establish procedures for disembarkation of patients to facilities capable of handling such conditions. PMID- 9867890 TI - Usefulness of the total iron binding capacity in the evaluation and treatment of acute iron overdose. AB - The ingestion of iron-containing products is a potential toxicologic emergency. The total iron binding capacity (TIBC) has been used widely as a predictor of end organ toxicity and a guide to the need for deferoxamine therapy. When the TIBC is greater than the serum iron concentration (SIC), it is held that no free iron is present to cause toxicity. The TIBC fails as a marker of toxicity for several reasons. First, the laboratory methods used to measure TIBC are inaccurate in the setting of iron overload. Second, the presence of deferoxamine, the antidote for iron poisoning, has been shown to make the TIBC measurement inaccurate. Third, TIBC measurements have been shown to be variable. Finally, studies and case reports demonstrate toxicity even when the TIBC is greater than the SIC. These shortcomings of the TIBC invalidate it as a predictor of toxicity in iron poisoning. PMID- 9867891 TI - A hierarchical outcomes approach to test assessment. AB - This report describes a hierarchical classification system for clinical endpoints in diagnostic technology assessment. The model requires classification of investigative outcomes into 1 of 6 categories, including technical efficacy, diagnostic accuracy efficacy, diagnostic thinking efficacy, therapeutic efficacy, clinical outcome efficacy, and societal efficacy. Evaluations at successive levels indicate a broader understanding of the test's value. The purpose of this classification system is to help readers of the medical literature consider various aspects of test performance, and to identify aspects of a test that require additional investigation. It is also designed to be used as a template for systematic reviews of diagnostic tests. In this review, examples from the medical literature are discussed to highlight important concepts from the hierarchy model and their relevance to emergency medicine. PMID- 9867892 TI - Evaluating bias and variability in diagnostic test reports. AB - Diagnostic testing is an important component of modern medical care. Unfortunately, many diagnostic tests are not rigorously evaluated before general application. Studies examining test characteristics often have methodologic flaws that impair their ability to provide reliable information on test performance. These flaws can introduce systematic nonrandom errors (biases) that distort measures of test accuracy. Other design errors can make it difficult to generalize the results of individual studies. These problems may enhance the apparent performance of poor tests while obscuring the performance of good tests, and they may result in the widespread use of tests with uncertain or limited efficacy. This article explores the ways in which studies of diagnostic test efficacy can be affected by bias and variability. PMID- 9867893 TI - Electron-beam computed tomography: shortening the short stay protocol. PMID- 9867894 TI - Are We ready to make "financial transfers" the standard of care? PMID- 9867895 TI - International emergency medicine and the recent development of emergency medicine worldwide. AB - Emergency medicine is being established as a unique and independent specialty throughout the world. Two major models of emergency care delivery exist in the world today: the Anglo-American and the Franco-German model. Most countries developing new systems of emergency care are following the Anglo-American model and are recapitulating the sequence of steps taken to establish the systems of emergency medicine in the United States. The most important step in the development of emergency medicine in other countries is the recognition that emergency medicine incorporates a unique body of knowledge requiring specialized practitioners or emergency physicians. A global network of international emergency medicine is assisting the development of emergency medicine worldwide and now includes international organizations, academic institutions, and individuals in countries where emergency medicine is mature and their counterparts in countries where emergency medicine is developing. The multilevel exchange of information through various modalities, such as international conferences, physician exchange programs, and print or electronic media, is playing a vital role in the search for internationally applicable systems of emergency care. PMID- 9867896 TI - Life-threatening diphenhydramine overdose treated with charcoal hemoperfusion and hemodialysis. AB - We describe a 35-year-old woman who ingested 16 g (approximately 20 mg/kg) of diphenhydramine with resultant hypotension requiring pressor support and marked QRS prolongation. After treatment with sodium bicarbonate failed to improve cardiac status, the patient underwent emergency charcoal hemoperfusion and hemodialysis with rapid improvement in the ECG findings within the first 40 minutes of treatment and with no further need for pressors. Major texts in toxicology state that diphenhydramine is unsuitable for hemodialysis or charcoal hemoperfusion. However, our case suggests that charcoal hemoperfusion may be appropriate therapy in cases of massive diphenhydramine overdoses when standard supportive measures fail. This case represents the largest documented diphenhydramine overdose that resulted in survival. PMID- 9867897 TI - Emergency department hemodialysis in a case of severe ethylene glycol poisoning. AB - A 36-year-old man with a history of depression presented to the emergency department after ingesting approximately 3,000 mL of ethylene glycol antifreeze in a suicide attempt. The patient's ethylene glycol concentration, 1,889 mg/dL, was higher than any level previously documented in the medical literature. Although his course was complicated by nausea, emesis, lethargy, metabolic acidosis, and kidney failure, the patient survived without persistent kidney failure or other chronic problems. Sustained hemodialysis and ethanol infusion were instituted in the ED, on the basis of the patient's history, before laboratory confirmation of the ingestion was obtained. PMID- 9867898 TI - Methylene blue by intraosseous infusion for methemoglobinemia. AB - Intraosseous administration of methylene blue may be an emergency alternative to intravascular administration. A 6-week-old female infant (3 kg) presented to the emergency department after a 1-week illness and appeared cyanotic and listless. Oxygen saturation by oximetry was 86% while the patient was receiving oxygen. Vital signs were blood pressure, 107/80 mm Hg; pulse, 190; respirations, 47; temperature, 39.0 degreesC. A metabolic acidosis and a methemoglobin level of 29.3% were present. After several unsuccessful attempts to establish intravenous access, an intraosseous needle was placed in the infant's left tibia. Methylene blue, 1 mg/kg, normal saline solution, and sodium bicarbonate were given intraosseously. The patient's oxygen saturation rose to 98% to 100%, and her cyanosis improved. Three hours later, her methemoglobin level was 8.2%. The child recovered uneventfully and was sent home after 3 days. Intraosseous administration of standard intravenous doses of methylene blue rapidly terminated the effects of acquired methemoglobinemia. PMID- 9867900 TI - Residents' perspective. PMID- 9867899 TI - Delayed toxic reaction following massive bee envenomation. AB - Massive bee envenomation can produce both immediate and delayed toxic reaction. Signs and symptoms of immediate toxic reaction are fatigue, nausea, vomiting, hemolysis, kidney failure, and disseminated intravascular coagulation. The label "delayed toxic reaction" refers to a patient who is asymptomatic after a massive bee envenomation, with normal initial laboratory results, but later demonstrates laboratory evidence of hemolysis, coagulopathy, thrombocytopenia, rhabdomyolysis, liver dysfunction, and disseminated intravascular coagulation. The subject of this case report, a 66-year-old man, was stung more than 125 times in an attack by Africanized bees. He was initially asymptomatic, except for pain, and his laboratory findings were normal. The first signs of his fatal multi-organ-system failure were not apparent until 18 hours after envenomation. This experience has led the Good Samaritan Regional Poison Center in Phoenix, AZ, to recommend a 24 hour hospitalization for pediatric patients, older patients, and patients with underlying medical problems who are asymptomatic or who are experiencing only pain after an envenomation of 50 or more stings. Such patients have an increased risk of tissue injury, which may be delayed and which may be more effectively treated if identified early rather than on 12- to 24-hour follow-up. All other envenomated, asymptomatic patients or envenomated patients experiencing only pain who become symptomatic or who belatedly exhibit laboratory values consistent with hemolysis, thrombocytopenia, rhabdomyolysis, liver dysfunction, kidney failure, and disseminated intravascular coagulation within a 6-hour emergency department observation period should be admitted. Intravenous fluids, blood products, dialysis, and other intensive measures should be initiated if necessary. PMID- 9867901 TI - Update on emerging infections from the Centers for Disease Control and Prevention. Hantavirus pulmonary syndrome--Colorado and New Mexico, 1998. PMID- 9867903 TI - Rapid sequence induction. PMID- 9867904 TI - On gun control. PMID- 9867905 TI - On firearm violence. PMID- 9867906 TI - Delayed hypoglycemia after ingestion of a single glipizide tablet. PMID- 9867907 TI - Delayed hypoglycemia after ingestion of a single glipizide tablet. PMID- 9867908 TI - Acute myocardial infarction after irradiation for Hodgkin's disease: a not so unusual case. PMID- 9867909 TI - [Experimental study on porous hydroxyapatite ceramics in repair of skull bone defect of rabbit]. AB - In order to investigate the possibility of porous hydroxyapatite ceramics (HAC) in the repair of skull bone defect, twenty-four rabbits were used. The bone defect model was created by operation to obtain a defect in parietal bone in a size of 1 cm x 1 cm. Filled the defect with HAC and methyl-methacrylate-syrene copolymer (MMAS) to fill the defect as control. At 1st, 2nd and 3rd months after operation, behavior of the rabbits was observed and then these animals were sacrificed and specimens were examined under microscope. Results showed as follows: after operation, behavior of all animals were normal. By histological examination, it was found that in HAC group, there were granulation tissue, fibrous tissue and newly formed vessels grew into the pores and the osteoblasts formed osseous trabeculae. There was no inflammatory cell infiltration. In the MMAS grafted asea, there was formation of fibrous membrane. It suggested that HAC might be a good material for bone substitute in repair of skull bone defect. PMID- 9867910 TI - [Experimental study of coral implantation in repair of skull bone defect in rabbit: histomorphometry of bone]. AB - In order to evaluate coral as a bone graft substitute in repair of bone defect, particulates of coral were implanted into skull bone defect of rabbit, 1.5 cm in diameter. Hydroxyapatite and blank were taken as controls. The rabbits were sacrificed at the second, fourth, eighth and twelveth weeks after the operation. The specimens were taken and performed histological examination and histomorphometry observation. Results were as follows: at the second week many multinucleus giant cells infiltrated. As time elapsed, the coral were progressively degenerated and new bone was formed to fill the defect. Up to the twelveth week, the coral degenerated completely and new bone formed in the center of the defect. Percentage of new bone was in defect was 36.9%. Compared with the controls, there were significant differences (P < 0.01). It was suggested that coral had good osteoconductility. Howevel, coral underwent rapid degeneration, it might result in inconplete repair of bone defect. PMID- 9867911 TI - [Clinical application of biological bone carrier in repair of long bone defect of femur]. AB - It is difficult to repair long defect of bone. Biological bone carrier (BBC) was one of the artifical bone substitutes. It was obtained from human or swine bone after a series of biochemical treatment. It had good histocompatibility. It had the same components and structure of bone, and its biological strength was samiliar to bone. In clinic, BBC was applied to repair of long defect of bone in two cases. The lengths of defect were 13 cm and 11 cm, respectively. After followed up for 2 to 3 years, it was found that the implanted BBC had been combined with the femur with new bone. It had the same metabolism and density as that of the normal bone. PMID- 9867912 TI - [Application of artificial humeral head in the treatment of tumor of upper end of humerus]. AB - In order to restore the function of shoulder joint in patient with tumor of upper end of humerus, artificial humeral head replacement was performed. The materials included resinene, nylon-6 and large molecular polyethylene from 1978 to 1993, 14 patients were treated. The tumors involved in this group were giant cell tumor, synviosarcoma, bone cyst, osteochondroma, osteoblastoma, osteofibrosarcoma, osteosarcoma, chondrosarcoma, parosteosarcoma and malignant giant cell tumor. After resection of the tumor, the artificial prosthesis was implanted with bone cement. After 1 to 16 years follow-up, functions of the shoulder joint were reserved in 86% of the patients. PMID- 9867913 TI - [Advances in the study of composite artificial bone]. PMID- 9867914 TI - [Morphological changes of intramedullary capillary circulation after fracture]. AB - To investigate the influence of injury of nutrient artery on the blood supply of inner 2/3 of cortical and intramedullary tissue, the experiment was carried out on twenty-four rabbits. Four rabbits were killed and perfused with Chinese ink in femoral artery to survey the blood supply of intramedullary vessels. Others were received fracture on tibia and sacrificed respectively on the third, eighth, eighteenth, twenty-eighth, and forty-second days after fracture. Chinese ink was also perfused through femoral artery on those rabbits. The relative density of capillary vessels were determined. The results were as follows: when the nutrient artery was injured after fracture, blood supply to the intramedullary tissue and the inner 2/3 of cortical bone was interrupted. The intramedullary vessels were increared and dilated. During the whole course of healing of fraeture, the density of intramedullary capillary vessels were higher than normal. It was concluded that the anastomosis between the epiphysis and metaphysis was important in the reconstruction of collateral circulation in healing of fracture. PMID- 9867915 TI - [The effect of phenytoin in healing of fracture of rabbits]. AB - It was reported that the systemic use of phenytoin could promote healing of fracture. In order to observe the effect of local application of phenytoin in the healing of fracture, the experiment was performed. Seventy-two rabbits were divided into three groups. Fractures were created on both radius of all rabbits. Group 1, intraperitoneal injection of phenytoin with a dosage of 50 mg/kg per day; Group 2, local use of phenytoin with a dosage of 40 mg/kg was injected in the fracture site every seventy-two hours, and Group 3, injection mormal saline of in the control group. Eight rabbits in each group were sacrificed in the 9th, 16th and 30th days after operation respectively. By X-ray excuiualtion, the healing of fracture was observed. Dry and wet weights of the callus were determined. After HE and Mallory's stain, the samples were examined under microscope. Results showed that both local and systemic use of phenytoin promoted healing of fracture. The effects of phenytoin in the two groups were the same and had no significant difference. PMID- 9867916 TI - [The effect of wumingyi chongji on BMP content in callus]. AB - A kind of Chinese herb medicine--Wumingyi chongji could promote fracture healing had been proved. In order to disclose the mechanism, a standard fracture model was produced in 50 healthy male Newzealand rabbits. The animals were randomly divided into experimental group and control group. Another 5 rabbits without operation act as nomal. By immunohistochemical study, the bone morphogenetic protein (BMP) content in callus was calculated by computerized interacive morphometry (CIM) in the 1st, 2nd, 3rd, 4th, 5th week after fracture. The results were: BMP in callus was much more than in nomal (P < 0.01); BMP in callus increased from the 1st week to the 3rd week, decreased from 4th week to 5th week, and was more in experimental group in the 1st and 2nd week than in the control group (P < 0.01). The highest BMP content appeared in the 2nd week in the experimental group, while in the 3rd week in the control group. It was concluded that Wumingyi chongji could promote osteoblast to synthetize BMP, BMP induce preosteoblast change into osteoblast. Thus the rate of fracture healing could be increased. PMID- 9867917 TI - [Improvement of technigue in restoration of stability of thoracolumbal spine]. AB - A new fixator for spine was designed to restore the stability of spine in improving the treatment of thoraco-lumbar dislocation. This instrument was composed of six nails, two longitudinal connecting rods and two transverse connecting rods. On the surface of the nails, there were furrows engraved, instead of screw thread. It fixed the spine through pedicle of vertebra from the posterior approach. The fixed vertebrae were limited between two vertebrae. From 1989 th 1995, twelve patients with fracture-dislocation of thoraco-lumbar spine were reduced with this instrument. After four years follow-up, it showed that the deformity was corrected and the spine firmly fixed. Compared with Dick's screws, it had the following advantages such as simple manipulation, increased strergth of nails and minimal damage to tissue during operation. PMID- 9867918 TI - [Total hip replacement in patients with steroid-induced femoral head necrosis]. AB - In order to evaluate the long-term effect of total hip replacement (THR) in patients with steroid-induced femoral head necrosis, 40 cases of 50 hips received THR patients were followed up for an average of 8.5 years after operation. Evaluation was carried act according to Harris score system. In these cases, average score was seventy-five points. Revision rate among them was 2% after four years and 18% after five to eight and a half years, with as overall rate of 20%. It was found that the main reason for revision was looseness of the prosthesis. In this follow-up, it showed that besides foreigen body reaction, abnormal osseous remodelling was the main factor in long-term failure of this arthroplasty. It was also proved that it was a good selection to perform cemented total hip replacement in younger patients, which could improve living quality. PMID- 9867919 TI - [Current status of the study of tissue engineering]. PMID- 9867920 TI - [Experimental study in vivo on implantation of autogenous tendon cells after combining culture with carbon fibers]. AB - In order to investigate the possibility of repairing injuried tendon with living artificial tendon, after combining culture, subcultured autogenous tendon cells with carbon fibers were implanted into the calcaneous tendon of rabbits. In different stages, the synthesis of type I collagen and their relevant morphological changes were observed. The results showed as follows: after implantation, tendon cells continued proliferating. Four weeks after implantation, tendon cells were detached from the carbon fibers and proliferated and produced collagen among the carbon fibers. The collagen fibrils were linked with each other to formed a dense structure. In the linkage site, the collagen fibrils originated from the implants joined to that from the ruptured end of the tendon, which meaned that the implant was healed with the recipient tendon. Observed under scanning electronic microscope, the tendon cells were lined among the carbon fibers evenly and in order, the collagen fibrils joined each other and formed an network, the fibrils were lined parallel to the carbon fibers. Under transparent electron microscope, the nucleolus were clear and organelle were abundant. PMID- 9867921 TI - [Relation between capsular contracture around breast implants and fibroblasts in breast reconstruction]. PMID- 9867922 TI - [TGF-beta 1 gene expression in the healing process of skin wound in rat]. AB - For observation of the change of transforming growth factor-beta 1 (TGF-beta 1) gene expression in the process of skin wound healing, the following experiments were performed. Sixteen Wistar rats were chosen. At each side of the rat's back, a 1 cm x 1.5 cm middle-thick skin wound was made. After 3, 6, 9 and 12 days, the specimens were taken from the wounds. For each specimen, half of it was used for RNA extraction, and underwent dot blotting; and the other half was frozen immediately and underwent in situ hybridization. The probes were dig-labeled PDGF BB cDNA probe and TGF-beta 1 probe. The results showed that TGF-beta 1 gene was expressed mainly in fibroblast, epithelial cell and capillary endothelial cell. The peak of TGF-beta 1 mRNA content was in the 6th day postoperatively. After that, the content of TGF-beta 1 decreased to normal. It was suggested that TGF beta 1 gene expression was in close relation with healing process. TGF-beta 1 may play an important regulatory role in the skin wound healing. PMID- 9867923 TI - [The effect of different way of using FGF on wound healing]. AB - To investigate the best way of using fibroblast growth factor (FGF) in wound healing, the following experiments were performed. Twelve Wistar rats were chosen. Four 1.5 cm x 1.5 cm middle-thick skin wounds were made in the back of each rat, 2 in each side, and labelled as number 1 to 4. Number 1 wound of each rat was used as control, only PBS was applied to the wound, 50 microliters per time, twice a day from the first day to 11th day. Number 2 wound was sustained medication group, 50 microliters 4 micrograms/ml FGF was applied twice a day from the first day to 11th day; Number 3 wound was early medication group, 50 microliters 8 micrograms/ml FGF was applied twice a day from the first day to 5th day; Number 4 wound was late medication group, 50 microliters 8 micrograms/ml FGF was added twice a day from the 5th day to 11th day. By day 4, 8, 12 and 16, the area of wounds were measured, and the healing time of each wound was recorded. The elastic fiber, collagen fiber and DNA content were measured by immunohistological method. The result showed that the elastic fiber, collagen fiber and DNA content in the groups of FGF used were more than those in the control group. The healing time of the control group was 14.4 days while that of the early meduation group was 13.4 days, late medation group was 13.5 days and sustained medication group was 12.2 days. It was suggested that FGF could accelerate the wound healing, and sustained use of FGF was the best way of giving the drug. PMID- 9867924 TI - [Effects of bFGF on succinate dehydrogenase level and oxygen consumption of skin flap in rats]. AB - To investigate the effects of basic fibroblast growth factor (bFGF) on necrosis rate, succinate dehydrogenase level and oxygen consumption of the skin flap, 18 Wistar rats were divided into 2 groups. Caudally based skin flap was raised on the back of each rat. Nine micrograms bFGF or normal saline with heparin was instilled under the flaps respectively after closure of the wounds. After 7 days, the necrosis rate of each wound was measured. The result showed that in bFGF group, the average necrosis rate of skin flap was 18.2%, less than that of the control group (37.14%). Succinate dehydrogenase content and oxygen consumption in bFGF group were higher than those of the corresponding sites in the control group (P < 0.05). It was suggested that the use of bFGF resulted in the decrease rate of necrosis of skin flap, and it maintained higher succinale dehydrogenase level and oxygen consumption. It was concluded that bFGF would probably be valuable for clinical use. PMID- 9867925 TI - [The study of epidermal growth factor in acceleration of skin wound healing]. AB - In order to investigate the function of epidermal growth factor (EGF), the following experiments were performed. Thirty white rats were chosen and divided into 3 groups. In the back of each rat, two 2 cm x 2 cm wounds were made bilaterally, the skin and subcutaneous tissue was removed. EGF were used in one of the two wounds randomly, while those without EGF usage in control. After 1, 2 and 3 weeks, the rats were sacrificed. The area of the wounds was measured, and the healing time of each wound was recorded. The results showed that the healing time of EGF group was 14.6 days while that of control group was 18.5 days (P < 0.01). Furthermore, the DNA, protein and hydroxyproline contents of EGF group were higher than those of the control group (P < 0.01). It was suggested the EGF could accelerate wound healing and shorten the healing time. PMID- 9867926 TI - [Basic fibroblast growth factor and nerve regeneration]. PMID- 9867927 TI - [Biological function and clinical use of fibroblast growth factors]. PMID- 9867928 TI - [Morphometric study of collagen fibers during healing following partial and complete section of extensor tendons in rats]. AB - In order to study the influence of severity of tendon injury on the morphology of collagen fibers during healing process of extensor tendons, 40 female Wistal rats were used for investigation. The rats were divided into 2 groups. Transection of the tendon of extensor digitorum longus was performed in one group, while partial section of the same tendon was performed in the other group. Morphometric analysis was undertaken on the 15th, 30th, 60th and 90th day after operation. The result was that there was no significant difference between the two groups both in distribution and diameter of collagen fibers on the 15th and 30th days (P > 0.05). However, there was significent difference between those on the 60th and 90th days (P < 0.05). It was concluded that the severity of the tendon injury could influence the morphology of collagen fibers during the late stage of tendon healing. PMID- 9867929 TI - [The effect of repair of paratendon in tendon healing]. AB - In order to investigate the effect of repair of paratendon in tendon healing, two different ways were performed to repair the transected extensor tendons of chick's toe. End to end suture of the extensor tenon was performed in group 1 while the paratendon was also repaired simultaneously in addition to suture of the tendon in group 2. Gross observation and histological examination were undertaken in the 3rd and 6th week after operation. The result showed, in group 1, extensive adhesion and irregular proliferation of fibroblasts was found in the 3rd week, severe adhesion and irregular arrangement of fibroblasts with less collagen fiber was found in the 6th week; while in group 2, smooth and regular "fusiform structure" was formed, slight adhesion and regular proliferation of fibroblasts were found in the 3rd week, adhesion disappeared and the structure of paratendon and tondon recovered in the 6th week. It was concluded that repair of extensor tendon and paratendon simultaneously could promote the intrinsic tendon healing and prevent tendon adhesion. PMID- 9867930 TI - [Close injury of the tendon at wrist]. AB - Because of the complicated causes and variable clinical signs, closed injury of tendons at wrist is difficult to diagnosis and treat. Twenty-six cases of tendon ruptur were reported. Among them, 11 cases were caused by bone fracture or dislocation, 8 cases were caused by rheumatoid synovitis, 5 cases were caused by synovial tuberculosis, and 2 cases caused by other. The pathogenesis and clinical signs were analyzed. Twenty-three cases were treated by tendon transfer and 3 cases were treated by tendon transplantation. By average follow-up of 16 months (ranged 6 months to 4 years), the results were as follows: the clip strength and both active and positive motion of fingers were restored in 19 caese, 75% of those were restored in 7 cases and 50% of those were restored in 2 cases. It was suggested that diagnosis, treatment and function rehabilitation should be carried out early, and tendon transfer or tendon transplatation were the method on priority. PMID- 9867931 TI - [Experimental study and clinical application of arterialized venous skin flap]. AB - In order to investigate the survival mechanism and the role of venous drainage in arterialized venous skin flap, 60 rabbits' ears were used for research and clinical application of the flap was performed subsequently in two cases. The rabbits were divided into 4 groups. Experimental group was standard arterialized venous skin flap, control 1 group was venous skin flap, control 2 group was arterialized venous skin flap with only one drainage vein and control 3 group was normal skin flap. The process of survival of the flaps was observed by hemodynamic and histological method. The results showed that there was no significant difference between standard arterialized venous skin flap and normal skin flap (P > 0.01). Two cases of arterialized venous skin flap survived completely. The conclusion were as follow: 1. the opening of collateral circulation between the veinlets was the main change of the microcirculation; 2. the blood flow of the graft was changed from unphysiological circulation to physiological circulation as the time elapsed and 3. amelioration of venous drainage was important in inproving the survival rate of arterialized vein graft. PMID- 9867932 TI - [Reform of the pedicled abdominal subcorium vascular-net flap and its clinical application]. AB - In order to resolve the shortcomings of traditional pedicled abdominal skin flap, the pedicled abdominal subcorium vascular-net flap was reformed and applied clinically. Twenty-eight cases with scar on hand or wrist were treated, including 20 males and 8 females. The age was ranged from 18 to 35 years old. The key point in the design was rotating 45 degrees of the flap from the primary site toward the pedicle. The ratio of the length to width of the flap was 1-1.8 : 1, and the wound of the donor site was covered by direct suture. Five to seven days later, all the flaps were divided and survived. The advantages of this flap were as follows: skin-grafting on the donor site was not necessary; the time needed for cutting the pedicle was shortened, and the flap is thinner than the traditional flap. PMID- 9867933 TI - [Repair of pulp defect of thumb by free palmaris brevis musculocutaneous flap]. AB - It is difficult to repair the pulp defect of finger with good function. Here reported two cases of pulp defect of the thumb which were repaired with free palmris brevis musculo-cutaneous flap. The flap was designed as the following: taken the line crossing the pisiform and metacarpo-phalangeal joint of the little finger as the longitudinal axis of the flap. The proximal end of the flap was at the level of pisiform and the distal end was the distal transverse palmar crease. The radial border was the radial side of the palmris brevis and ulnar border was the ulnar edge of the palm. The flap should not be larger than 6.5 cm x 2.5 cm. The flap was dissected with proper ulnar vessels and the accompanying arterial branch and superficial branch of the ulnar nerve. The flap was transferred to repair the pulp defect of the thumb. The vessels were anastomosed with the radial artery and cephalic vein at the snuffbox. The nerves were sutured to the digital nerve. Three months after operation, the defect was healed and the sensation of pulp was recovered. The donor area was painless and without sensation disturbance. The motion of the little finger was normal. The function of the thumb was restored. It was concluded that the palmaris brevis musculocutaneous flap is one of the best donor region to repair pulp defect of thumb. PMID- 9867934 TI - [The effect of insulin-like growth factor-1 on cyclic change of tendon cell]. AB - In order to investigate the effect of insulin-like growth factor-1 (IGF-1 on the cyclic change of tendon cell, the 6th generation of cultured tendon cell were selected, and 20 ng/ml IGF-1 was added to the medium. After 48 hours, the cells were determined by flow cytometer, as well as the control cells. The results showed that the time of G1 phase, DNA synthesis phase and G2M phase in IGF-1 group were 11.8 hours, 21.4 hours and 6.8 hours respectively, while those were 25.6 hours 22.6 hours and 21.8 hours respectively in the control group. It was showed that the time needed for G1 phase and G2M phase was shortened by IGF-1. PMID- 9867935 TI - [Implantation of allogenic osteoblast combined with calcium phosphate composites]. AB - The aim of this experiment was to study the osteogenesis in vivo of allogenic osteoblast combined culture with calcium phosphate composites. The osteoblasts were obtained by enzymatic digestion of periosteum from fibula subcultured to 13 generations, the cells were combined culture with hydroxyapatite and biphasic calcium phosphate. Subseguently, the composite was implanted into rabbits subcutaneously or intramuscularly. The blank material was implanted in the contralateral side as control. Four weeks later, all animals were sacrificed. All the implants were examined by gross observation, histological examination and EDXA. The results showed: 1. obvious ingrowth of connective tissue with very little inflammatory reaction; 2. new bone formation in the composites with deposit of Ca and P on the surface of osteoblast, but none in the blank materials; 3. no significant difference of new bone formation between the different sites of implantation or different materials, but those implanted intramuscularly had lamellae form of new bone while those implanted subcutaneously had only mineralization of extracellular matrix. The conclusion were: 1. the composites are biocompatible with prior osteogenesis property; 2. periosteal-derived allogenic osteoblasts obatined by enzymatic digestion could survive following implantation with bioactivity; 3. rich blood supply might be advantageous to new bone formation and its maturation. PMID- 9867936 TI - [Tissue engineered cartilage and its clinical use]. PMID- 9867937 TI - [In vitro experimental study of adriamycin-loaded chitosan drug delivery system]. AB - Chitosan is a kind of biological material with good histocompatibility and gradual biodegradability in vivo. It has no toxicity or side-effect. For its gradual degradation, chitosan and adriamycin were mixed and formed drug delivery system (DDS). The release test of DDS and exudant of DDS in inhibiting OS-116 were examined in vitro. The results were as following: the DDS could release adriamycin in slow and stable way. The SO-116 inhidition rate of the exudant of the DDS on the 1st, 20th, 40th and 60th day was 58.11%, 36.48%, 24.32% and 21.62% respectively. It was concluded that the drug delivery system was a slow release system. It could maintain the concentration of adriamycin in a certain level. It was also suggested that the chitosan was a good carrier for slow release of chemotherapeutic drug in local therapy for postoperative treatment of bone tumor. PMID- 9867939 TI - [Ten years of reconstructive surgery in China]. PMID- 9867938 TI - [Applied anatomy of osteo-periosteal flap pedicled with superior malleolar branch of anterior tibial artery]. AB - In order to investigate the blood supply of osteo-periosteal flap of lateral inferior part of tibia, 40 lower limbs of adult cadavers were observed. The result showed that the superior malleolar branch was the biggest branch on the lateral inferior part of tibia and served as the main blood supply to the above area. It originated from the anterior tibial artery, 3.1 +/- 0.8 cm above the intermalleolar line. During its way to the anterior border of the tibia, it gave out the ascending and descending branches. The ascending branch was along the anterior border upward and anastomosed with the musculo-periosteol branch of the anterior tibial artery at the level of 6.3 +/- 1.3 cm above the intermalleolar line. The decending branch was anastomosed with the anterior medial malleolar artery. For the anastomosis between the superior malleolar branch with the peripheral vessels, the osteo-periosteol flap could be designed at the lateral side of the lower part of tibia in size of 8-10 cm x 4-6 cm. This was a new donor area of osteo-periosteol flap for repair of non-union of bone in lower end of tibia or arthrodesis of the ankle joint. PMID- 9867940 TI - [Experimental study on nerve growth factor in protecting injured spinal cord]. AB - Thirty-five SD rats were divided into 3 groups, in which 5 rats were served as control. The other 2 groups, 15 rats in each received either NGF solution or normal saline. The injury at the level of T8 spinal segment of the rat in these two groups were made by dropping a weight of 10 g from a height 2.5 cm after a total laminectomly from T7-11 segments. A thin plastic tube was inserted into the subarachnoid space below the injured segments. NGF was introduced through the tube at intervals of 1, 2, 3, 4, 8, 12, 24, 48 hours in the NGF group, and normal saline in the normal saline group. At 4, 8, 24 hours following surgery, 5 rats in each group were sacrificed and the injured segments were resected for examination. The contents of water and calcium were measured by dry-wet method and atomic absorption spectroscopy. The result showed that total calcium and water contents in normal saline group were markly increased, however, the changes of these two parametere were not so prominent in NGF group. It was suggested that NGF play a role in protecting the spinal cord by maintaining the calcium level of the injured segment. PMID- 9867941 TI - [Studies on the changes of morphometry and neuropeptide of spinal neurons after peripheral nerve injury]. AB - To observe the change of morphology and neuropeptide in the spinal neurons in order to clarify the functional state after injury of peripheral nerves is especially in the late stage. Sciatic nerves were cut with their proximal segments in the preparation of a model of peripheral nerve injury. Combination of horseradish peroxidase retrograde tracing immunohistochemistry and computer image analysis the changes in the morphometry of the perikarya of ventral horn neurons of the spinal cord, the quantitative changes of substance P (SP). Calcitonin gene related peptide (CGRP) in dorsal horn and CGRP and choline acetyransferase (CHAT) in ventral horn of the spinal cord were examed. The results showd: (1) At the 3rd week after injury, swollen perikarya of the ventral horn neurons were observed, subseauently the swelling of perikarya was decreased tile the 6th week the neurons recovered to their normal size. At the 12th week the neurons were generally stable in their size, shortening of the dendrites was seen in 27% of the neurons. (2) The dendrites of the neurons progressively contracted till at the 12th week 53% of them were degenerated. The results of the 24th week were similar to the that at the 12th week. (3) CGRP in the ventral horn of the spinal cord was elevated to the highest point after 1 week of injury, that lasting for 4 weeks and 8 weeks later, the lever of CGRP returned to normal. From 20th to 24th week, there was no obvious changes of CHAT in the ventral horn of the spinal cord during observation. (4) SP went to the lowest point in the dorsal horn during 2-6 weeks, then recovered slowly, and beiny normal again after 16 weeks, however, CGRP was changed slightly. The results indicated that although a series of degenerating changes occurred in the neurons of the spinal cord during the late peripheral nerve injury, but the functional activity of the central meurons still was maintained at a certain level. PMID- 9867942 TI - [Influence of low energy He-Ne laser on spinal motor nerve cell]. AB - The object of this experimental study was to investigate the influence of low energy He-Ne laser on the motor nerve cells of the spinal cord. The experimental study included as follws: (1) Four rabbits were used in this experiment. The L5-6 spinal cord segment was irradiated by He-Ne laser percutaneously, the nerve velocity of the comon peroneal nerve was measured in order to determine the function of the spinal motor nerve cells when the peripheral nerve was intact. (2) The common peroneal nerve was transected on one side wothout repair, two weeks after laser irradiation, the grey mater of the spinal cord of L5-6 segment was procured for electronic microscopic examination. (3) The common peroneal nerve on the contralateral side was transected and followed by end-to-end anastomosis, and laser irradiation was done on the same spinal cord segment. Two weeks after irradiation, the nerve velocity of the common peroneal nerve and the toe expanding test were investigated. The results were: (1) the He-Ne laser can influence the spinal motor nerve cells function as expressed by latent rate when the peripherial nerve is intact. i.e. the nerve velocity is slower than mormal, and the amplitude is markedly decreared. (2) the change of the microstructure of the spinal motor nerve cells is comparatively slight in the 10 and 15 minutes groups. (3) the recovery of the nerve velocity and the toe expansion are more earlier in the 15 min. group. In short, the low-energy He-Ne laser can influence the function of the spinal motor nerve cells. PMID- 9867943 TI - [Influence of low energy He-Ne laser on regeneration of peripheral nerve]. AB - The purpose of this experiment was to elucidate the influence of the low-energy He-Ne laser on the function of regeneration of peripheral nerve. Forty-four rabbits about 2.5 kg body weight were used in the experiment. The animals were divided into 4, 8, 12, 16 weeks groups according to the observation period. Six animals were used in each irradiated group and in the control group 5 rabbits were used in each observation period. Regeneration of the axon and myelinc sheath, the latent rate of the common peroneal nerve, the conditions of the anterior tibital muscle and the toe expansion test were all observed systematically in both groups. The experimental results was: A few thin regenerated axon was seen at 4 weeks in the irradiated group, while in the control group it might be seen at 8 weeks, the P value was < 0.01. A low amplitude latent rate of the common peroneal nerve is determined at the peroneal side of the anterior tibial muscle in a few animal at 4 weeks of the irradiated group, and it is not observed in the control group, from 12 to 16 weeks. THe latent rate of the common peroneal nerve was the irradiated group than in the controlled, the P value was < 0.01. The regeneration of the myeline sheath was evident in the irradiated group, and also the slstion of the musdle fibers anterior tibial muscle was clearly visible than the controlled. 16 weeks postoperatively, the toe expansion test was normal in the irradiated group, while in the control group it was the same as seen at 12 weeks after operation in the irradiated group. Now it was certain that the low-energy He-Ne laser could promole the function of the spinal motor nerve cells and accelerate the axonal regeneration. PMID- 9867944 TI - [Neurotrophic factors and spinal cord injury. A review]. PMID- 9867945 TI - [Experimental study of peripheral nerve regeneration influenced inverted and non inverted distal nerve segments]. AB - The sciatic nerves of adult rats were sectioned bilaterally and the ends of the nerves were placed in silicone tubes. One side of the distal nerve segment was inverted and that of the contralateral side was non-inverted. After 2, 4, 6 weeks, the rats were killed and the specimens were removed for macroscopic, histologic and morphometric analysis. The results showed that either the inverted or non-inverted distal nerve segments had no influence on the number of the myelinated axons in the regenerated nerves, but the number and density of the myelinated axons was markedly diminished in the inverted distal nerve segments. PMID- 9867946 TI - [Clinical significance of changes of fibrillation potential amplitude following denervation of human skeletal muscle]. AB - To evaluate the value of clinical application of examination of fibrillation potential amplitude, 110 patients, 97 males and 13 females, were examined and only the maximum fibrillation potential amplitudes were recorded in 420 muscles. The results showed that there was no significant difference between sexes, ages and sides. However, significant difference was evident between the groups of different frequency (1+ to 4+). The fibrillation potential amplitude was maximum at 3 to 4 months after denervation and still remained at relatively high level for years in certain patients. No significant difference was showed between the time groups in incomplete nerve injuries. Surgery did not affect the course of fibrillation potential amplitude change. It was suggested that the muscle cells sustained their property for years after denervation in some patients, thus it might explain that satisfactory result could be obtained from operative repair in some late cases. The changes of fibrillation potential amplitude might indicate that the changes from muscle denervation was still reversible and might be more accurate than traditional method of examination. PMID- 9867948 TI - [Advances in the study of lengthening of peripheral nerves and its related subjects]. PMID- 9867949 TI - [Microsurgical restoration of foot tissue defects]. AB - From 1984 to 1994, 236 different types of traumatic defects of foot were repaired by microsurgical tissue grafting. They included simple cutaneous flap in 187 and composite flap in 49. Among the 236 different tissue flaps, vascularized flap was used in 97 and pedicled flap in 139. The 4 fore-foot and 6 heel defects were repaired by composite skeleted cutaneous grafts with scapula and vascularized febula respectively. After the follow-up from 1 to 10 years, all the grafted tissues were survived and healed well. The functions were satisfactory, and 186 patients had resumed their original works. The key to good function following repair was to maintaion the integrity of foot structures and the sensation of the sole and heel. PMID- 9867947 TI - [Treatment of nerve root avulsion of brachial plexus by nerve transfer]. AB - The results of nerve transposition for root avulsion of brachial plexas in 21 cases were reported. The methods of the nerve transposition were divided into four groups as followings: By transfer of phrenic nerve, accesory nerve, the motor branches of cervical plexus and intercostal nerves in cease; By transfer of phrenic nerve, accessory nerve and the motor branches of cervical plexus in 6 cases; By transfer of phrenic nerve and accessory nerve in 9 cases, and by transfer of phrenic nerve or the motor branches of cervical plexus or intercostal nerve in 5 cases. During operation, in 1 cases variation of the brachial plexus was found. Injury to the subclavian artery occurred in 4 cases and they were repaired, which is good for the blood circulation of the upper arm and nerve regeneration. Nineteen cases were followed up with good results. The overall excellent and good rate was 73.7%. It was considered that transposition of nerve should be a routine operation for the treatment of root avulsion of brachial plexus and the accompanied arterial injury should be repaired at the same time during operation, and the latter would be advantageous to enhance functional recovery of nerve. PMID- 9867950 TI - [Reconstruction of tendon sheath by autogenous vein graft in preventing adhesion]. AB - In order to prevent tendon adhesion following operation, autogenous great saphenous vein graft was used to reconstruct the tendon sheath. The operation was performed under microsurgical technique. This method was used to repair 23 tendons and 17 tendon sheaths. The early functional exercises were carried out after operation. Follow up from 10 months to 4 years, the prognosis was good except in 3 fingers, in which, the wounds were infected resulting the necrosis of the grafted veins and exposure of the repaired tendons. The details of the operation were introduced. It was emphasized that non-traumatic handling of the tissues was essential in preventing tendon from adhesion. PMID- 9867951 TI - [Reconstruction of cervical stability following old facet dislocation of cervical spine]. AB - A total of 12 cases of old facet dislocations of cervical spine treated between december 1988 and 1993 were analyzed in order to evaluate the efficacy of various surgical modalities. In this series, there were 8 males and 4 females, with ages ranged from 16 to 50 years old (averaged 37.8 years old). The duration from injury to admission to our hospital was ranged from 1 to 8 months (averaged 3.7 months). Dislocation levels were as follows: C3,4 in 1 case, C4,5 in 4 cases, C5,6 in 4 cases and C6,7 in 3 cases. Unilateral facet dislocation was in 7 cases and bilateral facet dislocation in 5 cases. Neurological status on admission was as follows: spinal cord and nerve root lesion in 5 cases, nerve root lesion alone in 5 cases and neurologically intact in 2 cases. Besides all facets receiving facetectomy and iliac bone graft, other four kinds of adjuvant treatments were used, including internal fixation by stainless wires laminae or spinous processes in 4 cases, Luque rod in 1 cases, anterior fibrolysis combined with posterior laminoplasty in 1 cases and sustained skull traction without internal fixation in 6 cases. The reduction efficacy from postoperative stustained skull traction was better and the stainless wires fixation ranked the next. The patients only suffering from the nerve root lesion recovered better, but those who had spinal cord combined with nerve root lesion recovered badly. In conclusion, for the treatment of old facet dislocation, it is necessary to resect the facet and graft with iliac bone. PMID- 9867952 TI - [Reconstruction of lacrimal passage in repair of oblique facial cleft]. AB - For the reconstruction of lacrimal passage in obligue facial cleft, three methods were performed in 4 cases. According to the difference in local pathology canaliculorhinestomy, transfer of vein or oral mucosa, or modified dacryocystorhinostomy was performed. The patients were followed for 6 to 12 months with satisfacting results. PMID- 9867953 TI - [Experimental study on biological characteristics of tenocyte and fibroblast in rabbit]. AB - In order to study the biological characteristics of tenocyte and fibroblast, the former was obtained from rabbit's tendon, and the latter from rabbits's skin. Both cells were cultured according Heuderson's method. The cell morphology, strapping and expanding time, and the type of collagen fiber synthesized in culture were observed. The results showed that the strapping and expanding time of fibroblast was faster than that of tenocyte. The cellular arrangement of fibroblast was irregular, but that in tenocyte was regular. Type I and III collagen of fibers were found in cultured fibroblost while only type I collagen fibers were found in culture of tenocyte. The tenocyte and fibroblast could be identified individually by strapping and expanding time, arrangement of cells and type of collagen fiber synthesized. PMID- 9867954 TI - [Effect of biological electric stimulation on free muscle transfer]. AB - The rectus femoris muscles of rabbits were used as muscle model. The electrical stimulation which resembled the normal motor-unit activity was used to observe its effects on free transferred muscle. After three months, the moist muscle weight (MW), its maximum cross-section area, its contractility and its histochemical characteristics were examined. The results showed that the function and morphology of the muscles were well preserved. These findings might encourage its clinical application. PMID- 9867955 TI - [Experimental study prevention of reperfusion injury of skeletal muscle by local hypothermia and its clinical application]. AB - Local hypothermia as a preventive method to reperfusion injury of skeletal muscles was studied. Sixteen Japanese rabbits were divided into four groups at random. Before the tourniquet was inflated, a cold gel pack was applied to the right hind leg of each rabbit for 15 minutes to produce local hypothermic condition, without application of tourniquet the left hind limb was under local hypothermic condition as a control. The duration of tourniquet ischemia was 4 hours, and then reperfusion for one and two hours in the A and B groups respectively; in the C and D groups the duration of ischemia was 5 hours, and reperfusion for one and two hours, respectively. The muscle temperature averaged 16.6 degrees C with a needle thermocouple in the hind limb under local hypothermia. The serum K+, LA, SOD, LPO were determined from bilateral femoral veins, and electron and light microscopic studies of sural muscles were done in the post-reperfusion period. It was found that the K+, LA, LPO were lower than that of the control groups (P < 0.01), but SOD was higher than that of the control group (P < 0.01). Electron and light microscopic studies showed sight but reversible damage of muscular structure with the possibility of in the hypothermic groups cell regeneration. Basing on this experimental results, this method was applied in 45 cases reparative and reconstructive surgery of limbs. The duration of application of tourniquet averaged 2 hours and 57 minutes, the longest being 4 hours and 31 minutes, when the muscle temperature had reduced to 22.4 degrees C. There were no postoperative complications associated with this technique. Local hypothermia appeared to be a safe and effective method of decreasing the reperfusion damage after ischemia. PMID- 9867956 TI - [Adipose tissue transplantation and plastic surgery: review of basic research]. PMID- 9867957 TI - [The function of nitric oxide in the necrosis of avulsed skin flap in domestic pig]. AB - To investigate the function of nitric oxide (NO) and nitric oxide synthetase (NOS) inhibitor, N-nitro-L-arginine methyl ester (L-NAME), the skin avulsion model was made in the lower extremity of pig. The methods of measurement of size of the survived flap, weighing, immunocytochemistry and hybridization in situ were employed, so that the survival surface area of flaps, tissue wet/dry weight ratio, NO content in the serum, gene expression of NO and NOS content in the flap tissue were determined, respectively. The results showed that the early gene expression of NOS was increased as well as the NO content and tissue wet/dry weight ratio (P < 0.01). After L-NAME was applied introvenously, the NO content and tissue wet/dry weight ratio were decreased (P < 0.01), and the survival surface area of flaps was enlarged (P < 0.01). It could be concluded that the NO might play a role in the development of the pathological changes as early congestion, edema and secondary necrosis in the avulsed skin flaps. The early application of L-NAME could do some good to the avulsed skin flap and protect it from further necrosis owing to the presence of NO. PMID- 9867958 TI - [The effect of far infrared rays on the survival of randomized skin flap in the rat: an experimental study]. AB - In order to observe the effect of far infrared rays on the survival of skin flap, the following experiment was performed. Forty-eight SD rats were selected and divided into two groups. The rats received 0.3 w/cm2 radiation twice a day from 3 days before operation to 5 days after operation in the experimental group, while in the control group the rats received none before or after the operation. The flap was designed as 2 cm x 6 cm in the back of the rats with the pedicle caudalward. The microcirculatory changes of the flap were observed, and the survival area of the flap was calculated. The results showed that either in the proximal or in the distal part of the graft, in the experimental group, the mean opening rate, diameter and the flowing velocity of the microvessels were significantly higher than those in the control group (P < 0. 05). The mean rate of survival area of the experimental group (80.5%) was also higher than that of the control group (62.7%) (P < 0.01). It was suggested that radiation with far infrared rays could dilate the microvessels, improve the flap microcirculation, therefore, enhance the survival of the randomized skin flap. PMID- 9867959 TI - [Histological change of keratinocyte in full-thickness skin autograft and its effect on hyperpigmentation of the graft]. AB - To understand the reason of hyperpigmentation following full-thickness skin autograft, 54 guinea pigs were selected as skin autograft models. The changes of density of epidermal keratinocyte (KC) and the amount and distribution of melanin in the epidermal KC were observed by histological, histochemical and autoradiographic techniques. The results showed: (1) The histological changes of KC were in fact a wound-repair process; (2) The increased melanin in KC coordinated with "multi-overlapping screen" effect, a result of multiple layers of "supranuclear cap", was the direct cause of hyperpigmentation of the graft and (3) The hydrolytic disturbance of melanin in KC and the longer life span of the KC were probably the important reasons for the increase of melanin in the epidermis. PMID- 9867960 TI - [Rapidly expanded skin tube studied by histological and stereological observations]. AB - In order to investigate the histological and stereological change of the rapidly expanded skin tube, 15 Japanese Long Ear Rabbits were divided into 3 groups: (1) Rapid expansion group, injecting once a day for 7 days; (2) Routine expansion group, injecting every 3 days for 21 days and (3) Control group, no expander implanted. The results showed that: in the rapid expansion group, the thickness of the epidermis and dermis were both increased, fibroblast proliferation was active, a large amount of new capillaries was observed, the collagen fiber was in loose type, and the mitochondria and rough endoplasmic net of epithelial cell were increased. In the routine expansion group, similar changes were observed. The vascular density and volume fraction were significantly higher in rapid expansion group than those in the control group (P < 0.05). It was concluded that the rapid expansion of skin tube was feasible. PMID- 9867961 TI - [Clinical application of retrograde island flap carrying plantar metatarsal arteries as pedicle]. AB - The skin and soft tissue defects or ulceration of the wight-bearing part of the sole was difficult to repair with medial plantar island flap, but would be treated with retrograde island flap carrying plantar metatarsal arteries as pedicle. Ten flaps were applied in 9 patients. They had either indolent ulcer or skin defect secondary to excision of painful corn or callosities of the front part of the sole. The flaps were 3 cm to 5 cm long and 3 cm to 4 cm wide, and they all survived following retrograde transfer. The patients were followed up for 1 to 10 years. It was found that the patients could bear weight on the operated foot and could walk without pain or lameness. The flaps were resistant to abrasion from long-time walking. It was concluded that this kind of flap was best suitable to repair the ulcers and defects over the front part of the sole despite there were some minor shortcomings such as the size of the flaps available was small and the donor site required split skin graft for coverage. PMID- 9867962 TI - [Pedicled thoraco-abdominal skin flap applied in early repair of deep ulcer of upper extremity resulting from snake-bite injury]. AB - The ulcer resulting from snake-bite injury was characterized by deep and wide tissue necrosis and secondary infection. The patient was at high risk of loss of function of his extremity. From 1989 to 1996, 16 cases with deep ulcer of the upper extremity resulting from snake-bite injury were treated with different types of axial thoraco-abdominal skin flaps, depending on the location of the ulcer. Thoraco-umbilical skin flap was used in 2 cases, lateral thoracoabdominal skin flap in 1 case, iliolumbar skin flaps in 5 cases, lower abdominal skin flaps in 6 cases, lower abdominal divided foliated skin flap in 1 case and Y-shaped hypogastric skin flap in 1 case. Reparative operation was carried out within 3 weeks after injury and primary repair were undertaken in 6 of them. The pedicles were divided in 17 to 24 days after operation. Twelve flaps survived. Three of the 6 flaps had mild signs of inflammation which disappeared after administration of antibiotics. One had necrosis of the distal part of the flap, and was healed with split skin graft. This type of skin flap was an ideal method for the treatment of snake-bite injury of the upper extremity. PMID- 9867963 TI - [Clinical use of bilateral cervico-thoracic skin flaps]. AB - In order to study the clinical efficacy of bilateral cervico-thoracic skin flap on repairing the contracture of the burn scar of the neck, 66 flaps were used in 33 patients from 1983 to 1995. The size of the flap ranged from 5 cm x 6 cm to 8.5 cm x 15 cm. The donor site was covered with split skin graft. The ratio between the length and the width of the flaps should not exceed 3:1. Fifty-nine flaps survived completely, but 7 had necrosis of small area which was healed without any influence on the function and appearance. The operative technique of the bilateral cervico-thoracic skin flaps were reported. The advantages of this type of skin flap and its applied anatomy and the postoperative care were discussed. In the repair of the cicatritial contracture deformity of the neck, it was important to define whether the skin defect was located in the submandibular, anterior cervical or anterior thoracic region, thus appropriate type of repair could then be given accordingly. PMID- 9867964 TI - [The application of transposition of pectoralis major muscle or myocutaneous flap in the repair of long-standing wound over the mandibulo-cervico-thoracic area]. AB - The pectoralis major muscle or myocutaneous flap is characterized by its easy dissection and free rotation, as well as abundance of blood supply and tissue mass of the flap, so that it was often used in repairing the wound over the mandibulo-cervico-thoracic area. From 1989 to 1996, 11 cases with long-standing wound of this area were difficult to repair with the traditional method, but were treated with transposition of this type of myocutaneous flap. The myocutaneous flap was applied in 8 case and muscle flap in 3 case. The flap area ranged from 12 cm x 8 cm to 26 cm x 18 cm. All the flaps survived, and excellent results were achieved. The technique and design of the operation were reported in details. It was concluded that the process was easy to perform, and the flap had abundant blood supply and high resistance to infection. PMID- 9867965 TI - [Effects of hyperbaric oxygen on the skin graft survival]. PMID- 9867966 TI - [Advance in the study of restoration of sensation in skin flaps]. PMID- 9867967 TI - [Progress in the clinical uses of fasciocutaneous flap]. PMID- 9867968 TI - [Manufacture and application of a new composite allograft]. AB - It is in urgent need clinically to look for an ideal substance for the coverage of burn wounds owing to shortage of autografts or allografts. After the cadaveric skin was extracted with acetic acid, salted out with NaCl and freeze-dried to prepare a porous collagen membrane. The membrane was seeded with allo-epidermal cells and allo-fibroblasts on its two sides, respectively, and then was cultured to achieve an artificial composite allograft. The artificial composite allograft was then transplanted onto ten severly burned wounds. One-year follow-up showed satisfactory results and the histological examination confirmed that the composite allograft could improve the adherence and growth of the epidermal cells and was helpful for blood vessels and healing of non-inflammatory connective tissues in the wounds. PMID- 9867969 TI - [Morphological observation of combined-culture of tendon cell or fibroblast of rabbit with artificial materials in vitro]. AB - In order to investigate the compatibility and growth between the tendon cell or fibroblast of rabbit and artificial materials, the combined-culture of the two cells with the carbon fiber, terylene and chitin was observed respectively. Results showed as following: in vitro, the compatibility of carbon fiber with these two cells was well, cell-adhesion ability was good as well. Few cells grew on terylene. Chitin inhibited the growth of either cells. No matter the tendon cell or the fibroblast, the amount of cells adhering on the carbon fiber was far more than that on terylene or chitin. When the three materials were interlaced together, the collagen fibers produced by the cells were arranged in direction parallel to the carbon fibers. As the time elapsed, the cells on the carbon fiber distributed evenly and enveloped the material in network-like fashion, this suggested that carbon fiber was a good material for producing living artificial tendon and ligament. PMID- 9867970 TI - [Bowel elongation technique by using omentum as vascular pedicle in beagle dogs]. AB - Basing on the establishment of an isolated bowel segement (IBS) in Beagle dog by omentoenteropexy, the possibility of using omentum as vascular pedicle in bowel elongation was investigated. Five young Beagle dogs (3 females, 2 males) with a body weight of 5-9 kg were used. A longitudinal incision (15 cm long) was made down to the seromuscular layer of the jejunal segment on the antimesenteric border, 20 cm distal to the duodenojejunal fold, the mucosal layer should never be injured. Then the free margin of the omentum was attached to the seromuscular border of the jejunum by interrupted sutures. One dog died from volvulus and necrosis of the bowel 3 days after operation. Seven weeks later, the mesentery was ligated in one dog and the bowel lengthening procedure was carried out in the other 3 dogs. The procedure consisted of longitudinal splitting of the isolated bowel segment anteroposteriorly, thus two separated segments of bowel were obtained, each received its own blood supply from either omental or mesenteric blood vessels. Either of the longitudinally splitting segments was rebuilt into one intestinal canal by 3/0 non-traumatic sutures. Then isoperistaltic end-to-end anastomosis of these two isolated segments was carried out, and an jejunostomy was established. The free jejunal ends were rejoined together to restore the continuity of the bowel lumen. Two weeks later, the blood supply of the experimental bowel segment was observed. It was shown that the IBS and elongated bowel segment (EBS) appeared viable, and there was good collateral circulation between the bowel segment and the omentum. The regeneration of lymphatics was observed from injection of methylene blue. It was concluded that a viable IBS and EBS could be established by using omentoenteropexy technique. The application of the this model in the management of short bowel syndrome needed to be further investigated. PMID- 9867971 TI - [Experimental study of the effect of motion on repairing defect of articular cartilage following autogenous periosteal graft]. AB - In order to investigate the effect of motion on repairing articular cartilage defect following autogenous periosteal graft, sixty adult rabbits were divided randomly into three groups: out-cage motion (OCM), in-cage motion (ICM) and immobilization (IMM). A defect of the articular cartilage, 1 cm x 0.5 cm in size, was made in the patellar-groove of femur of each hind limb. Free autogenous periosteal graft from the proximal tibia was sutured on the base of the left defect, while the right limb was served as control. The animals were sacrificed at 4, 8 and 12 weeks, respectively, after operation. The regeneration of the cartilage implanted was observed through gross, histology, histochemical assay and electronic microscope. The influence of different amount of motion on the chondrogenesis from the periosteal implant was also compared. The result showed that the hyaline cartilage produced from periosteal implant could be capable to repair full-thickness of articular cartilage. From statistical study, there was significant difference between OCM and ICM groups (P < 0.05), ICM and IMM (P < 0.05) as well as OCM and IMM (P < 0.01). It was suggested that the periosteal graft was effective in repair of defect of articular cartilage and the amount of motion was important for chondrogenesis. PMID- 9867972 TI - [The study of enhancement effect on wound healing from acid/heat stable peptides derived from pig serum]. AB - It was reported in this article that a preparation of acid/heat-stable peptides (AHSP) from pig serum with a molecular-weight less than 18 ku a without antigenity and toxicity could exert enhancement effect on wound healing. Two pieces of polyvinyl alcohol (PVA) sponge implanted in rat dorsal subcutaneous pouchs of 20 mice were selected as the wound model. The subcutaneous pouch having one piece of sponge was taken as the experimental group and the other as the control. Injection of 50 microliters of such peptide preparation into the test sponge was performed once a day from the time of injury on for 5 consecutive times, while 50 microliters of BSA (5 mg/ml) into the control sponge in the same way. The levels of total DNA, protein and hydroproline in AHSP-treated sponge were observed significantly higher than those in the control sponge on the 7th and 10th days after wounding (P < 0.05). No significant difference was seen on the 14th postinjury day (P > 0.05). The effect of AHSP on proliferation of wound fibroblast cultured in vitro was also detected. In conclusion, such peptides derived from pig serum had the activity to accelerate wound healing without resultant excessive healing and its direct stimulation of the proliferation of wound fibroblast was probably one of the way which AHSP exerted its action. PMID- 9867973 TI - [Telescopic adhesive anastomosis of small blood vessel applied in formation of arteriovenous fistula for hemodialysis]. AB - The formation of an arteriovenous fistual for dialysis by routine interrupted sutures anastomosing the vein and artery is difficult to perform and time consuming. A new method, telescopic adhesive anastomosis was studied and applied in 10 hemodialysis patients, who were in need of an arteriovenous fistula. The external diameter of the vessels anastomosed was 2.40 +/- 0.20 mm (radial artery) or 2.40 +/- 0.35 mm (cephalic vein). After thorough debridement of the vascular ends, the arterial end was put in the venous lumen. In order to fix the telescopic vessels, two stitches were applied 180 degrees apart from each other and tied. Each stitch was inserted from vein (penetrating the whole wall) to artery (just through the adventitia and partial thickness of the media vasorum). The distance from the stitch to the edge of the vein was 0.5 mm, and that of the artery was approximated to the external diameter of the vessle. The medical adhesive was then applied for sealing the anastomotic adventitia. Ten seconds were given for the solidification of the adhesive. The patients were followed up for 8 months. The patency rate was 100%, and the rate of blood flow was more than 300 ml/min (measured by ultrasonography). It was shown that this method could be managed easily and quickly, and the so-formed fistula would fulfill the need of hemodialysis. PMID- 9867974 TI - [Advance in the study of post-wound effects of arginine]. PMID- 9867975 TI - [Uses of artificial implants in the repair and reconstruction of craniomaxillofacial area]. PMID- 9867976 TI - [The three-dimensional titanium miniplate rigid fixation in the treatment of fracture of maxilla]. AB - The three-dimensional (3-D) Titanium miniplate system is one of the newest internal rigid fixation for the maxillo-mandibular surgery in recent years. The geometry of the plate assures a good stability in the three-dimensions of the fracture sits. Thirty patients suffering from maxillo-mandibular fractures had been operated on using the 3-D Titanium miniplates since 1991. The group of the patient included 16 cases of mandibular fractures, 8 cases of maxillary fractures and 6 cases of maxillo-mandibular fracture. All of the patient had very good result without any complication. The follow-up was 6 months to 3 years. The 3-D Titanium miniplate as an ideal easy to use, good resistance against torque forces and, compact forms of the miniplate were the some of the advantages. Clinical examples had been provided to illustrate the actual usage of the 3-D miniplate in the field of the surgical treatment for the maxillo-mandibular fracture. It is the author's opinion that the internal rigid fixation by 3-D Titanium miniplates is a promising method of treating fractures of the maxilla and is to be better tolerated by the patients and surgeons. PMID- 9867977 TI - [Histological study on the using of autogenous costal perichondrium graft to repair the cartilage of condylar process of mandible]. AB - In order to observe the histological changes of the autogenous perichondrium graft from rib in the repair of injured articular cartilage of the condylar process of mandible, 50 rabbits were used, in which 15 were served as control. The articular cartilage with its subchondral bone were resected and an autogenous graft of costal perichondrium was sutured onto the raw surface of the condylar process, and in the controls, only the articular portion of the condylar process was resected without the application of autogenous costal perichondrium graft. The morphological changes of the newly formed cartilage during the process of its development were investigated by hiostological and autoradiog aphic techniques. The result revealed that 10 days after operation, the graft had increased in thickness and was richly populated form the proliferation of mesenchyme-like cells. Twenty to thirty days later, the chondrocytes were matured and the newly formed cartilage had covered the bony surface of mandibular condyle. At 60 days, the newly formed cartilagenous joint surface became glossy, and the morphology and arrangement of cells tended to be regular simulating the morphology of normal articular cartilage. From the experiment, it could be concluded that (1) The autogenous perichondrium graft placed on the condylar surface of mandible could form new articular cartilage which was similar in tissue morphology to the normal condylar cartilage. (2) The process of development of newly formed cartilage was similar to that of the normal cartilage. (3) The motion and loading on the joint could promote the formation of new cartilage and undergo biological reformation, gradually resulting in normal joint morphology. On this basis, the clinical application of autogenous perichondrium graft to repair injured cartilage of the condylar process of the mandible was feasible. PMID- 9867978 TI - [Frozen-preserved tarsus of eyelid from new born in the repair of palpebral defect]. AB - In order to repair palpebral defects resulted from various causes, frozen tarsus of eyelid from newborn was used. From Jan. 1993 to Feb. 1995, the frozen preserved tarsus of eyelid from new-born was used to repair the palpebral defect in 10 cases. These defects were resulted following operation in traumatic defect in 5 cases, tramatic defect in 3 cases and congenital defect in 2 cases. After 3 month to 3 years follow-up, no refection reaction was found and no complication was occurred. The external appearance following repair was good. The overall successful rate was 100%. It was suggested that the frozen preserved tarsus from new-born was a safe and reliable material in the repair of palpebral defects. PMID- 9867979 TI - [Repair of cornea-sclera defect by autogenous sclera graft from the same eye]. AB - In order to rescue the serious cornea-sclera defect, normal piece of a sclera from the damaged eye was used to serve as a donor material to repair the cornea sclera defect. Eighteen cases were treated by transplantation of sclera graft with the operation to decompress the intra-ocular tension and to perform an artificial pupil from 1979 to 1994. Because the ocular-store seriously lacks in mostly internal locality, in the 18 cases, laceration of avulsive in 5 cases, severe corneal fistula complicated to other corneal diseases in 8 cases, locolized staphyloma of aornea in 2, laceration of sclera fistula in 1. The resulte were: The repairs were all healed up in an average of 20 days. The tension returned to normal and the vision was improved. The scleral graft was fused with the cornea, and the white area form the graft tended to reduce in size gradually with increase in transparency. There was no sign of iritis and rejection reaction. Follow-up of 3 to 36 months showed that the grafting operation was simple and an easy method, at the same time, once gained effect if would prevent or cure blindness. PMID- 9867980 TI - [Three kinds of standing-materials for concha in reconstruction of ear]. AB - In the reconstruction of the concha, standing support was necessary. From 1984 to 1996, 33 cases of aurical defects were admitted. Three standing-materials were chosen, and they included carved autogenous cartilage, heterogenous concha cartilage and steel wire work silicon-wires with silastic rubber tube. After expansion of the postauricular skin by tinsion expander, the standing-material was enveloped and total ear reconstruction or repair was performed. After followed up for average of 3.5 years, of the twenty-two cases, 16 had a satisfactory result, unsatisfied in 3 and failure in 3. It was concluded that the outcome of autogenous concha cartilage as a standing-material was good. The other two materials if used should be very carefully. PMID- 9867981 TI - [Intraoperative extra strong electrical stimulation in the treatment of peripheral nerve injury]. AB - In order to enhance the therapeutic effectiveness of peripheral nerve injury, intraoperative extrastrong electrical stimulation was used in peripheral nerve surgery. In 16 cases of incomplete peripheral rnerve injuries or poorly regeneratedn erves, continuous intraoperative electrophysiological monitoring was used for guidance of neurolysis. Meanwhile, extrastong electrical stimulation was applied. The latency and amplitude before and after electrical stimulation were recorded and the extent of improrement was compared. In all cases, the latency and amplitude were improved after neurolysis and electrical stimulation. Clinical follow-up also showed that the function of corresponding innorvated muscle was improved. Continuous intraoperative extrastrong electrical stimulation could be used as an practical measure to increase the effectiveness of peripheral nerve treatment. PMID- 9867982 TI - [Treatment of cicatricial deformity of neck with local skin flap]. AB - In order to correct the dysfunction of head and neck with scar contracture, since 1980, sixty-two cases were undertaken the operation by using local skin flap to repair the soft tissue defect after scar resection. The skin flaps included pedicled delto-thoracic skin flap in 26 cases, cervico-thoracic skin flaps in 25 cases, cervico-shoulder flaps in 6 cases, pedicled vascularized extralong delto thoracic skin flap in 4 cases and free parascapular flap in 1 case. Sixty cases had total survival of the flaps and 2 flaps had partial necrosis. After 1 to 10 years follow-up, the appearance and function of neck were excellent. It was suggested that grafting local skin flap was a good method to treat cicatricial deformity of neck especially using the skin flap with pedicle and vascular bundle. PMID- 9867983 TI - [The problems in the use of superficial cervical artery skin flap and it's relevant solution]. AB - Superficial cervical artery skin flap is widely used in clinical practice. In order to inprove the outcome of the flap in clinic, eleven cases of skin defect of scalp who were treated with the flap was discussed. After operation, the donor area healed but there was no hair growth on recipient area. Among them, six cases occurred partial necrosis of skin flaps. In order to avoid these problen, the relevant solution discussed as follows: 1. Handle well the pedicle of the skin flap to prevent the interference with venous returm. 2. Adhere strictly to indications. 3. Apply skin expander to obtain "extra" skin, then carryout the tranfer of skin flap and 4. Better use the skin flap with residual hair. PMID- 9867984 TI - [A preliminary report on clinical application of biopoly ester ligament]. AB - Sixty-one cases of fracture or dislocation were treated with France-made biopolyester ligament. They consisted of 18 cases of fracture of patella, 13 cases of fracture of olecranon, 8 cases of fracture of distal clavicle, 10 cases of dislocation of acromino-clavicular joint, 6 cases of separation of lower tibio fibular joint, 3 cases of rapture of cruciate ligament and 3 cases of fracture of upper third of ulna with dislolcation of radial head. The follow-up period lasted from 3 to 12 months with an average of 7.3 months. The result showed that of the 56 follow-up cases, 49 were excellent and 7 were satisfactory. There was no redisplacement occured in this group. The Biopolyester ligament was believed to be a good and safe material for fractures or dislocations and was of good strength of extension and easy to use. PMID- 9867985 TI - [Design of noninvasive steel miniplate for tenodesis and its clinical application]. AB - To fix the ligaments, tendons or other transplantation materials by passing through bone channels was commonly used and a relialde method in the reconstruction of joint stability. Usually an additional incision was necessary on the side of the outlet of the bone channel, that the screws, plates or U shaped staple could be applied. An instrument was designed that the steel miniplate could be fixed to the bone by noninvasive method. This technique was applied in 20 cases in which anterior cruciate ligament in 14, posterior cruciate ligament in 4, and collateral ligaments in 2 with carbon fibers. The result was successful in all patients. It was shown that it gave simple manipulation, reliable fixation and no need to have an additional incision, and it could lessen patients' suffering and lead to satisfactory clinical result. PMID- 9867986 TI - [Application of pedicled parumbilical axial skin flap in the repair of skin defect of upper extremity]. AB - The parumblical axial skin flap contained leap the parumbilical perforating branch of the inferiorle expigastic artery in its pedicle. It was designed to treat the scaring wounds of the upper limb in 5 patients with success. In the five patients, the scar on the dorsal aspect of the hand in 2, over the elbow in 1, on the palmar aspect in 1 and wound over the elbow in 1. The size of the flap ranged form 23 cm x 10 cm to 16 cm x 7 cm. The pedicle of the flap was made a tube. The limb was immobilized by plasten of paris after operation. All of the flaps were survived. The pedicle was divided at the hird week postoperation. The vascular supply of the flap was constant. The operation was also easy. This flap was feasible for the repair of the scar or wound of the hand, wrist, forearm, and, especially the elbow. PMID- 9867987 TI - [Dumbbell-shaped double-leaf flap for the repair of penetrating wound of palm with tissue defect]. AB - The penetrating wound of palm with the defect of tissues was usually caused by blunt crushing of gun-shot injury. It was difficult to treat clinically. In order to repair the skin defect of palm and dorsum in one operation, 2 kinds of dumbbell-shaped double-leaf flap were desigened. From February 1993 to March 1996, 5 cases with penetrating wound of palm were treated by this method in which the interosseous posterior artery flap in 4, and the superficial epigastric flap in 1. There were 4 males and 1 female with the ages ranging from 15 to 47 years old. The results were as follows: total survival of the flap in 4 cases and partial necrosis of skin margin of the distal leaf in 1 case. The appearance and function of the flap were satisfactory after a follow-up from 4 months to 3 years. It was concluded that the interosseous posterior artery flap had a long vascular pedicle with its constant anatomical distribution, and the 2nd stage of division of the pedicle of the flap was not necessary. It was a perfect flap in repairing the penetrating wound of the palm with small skin defect. The superficial epigastric flap was cumbersome in appearance and was necessary to divide the pedicle of the flap in 2nd staged operation. It was suitable in repairing the penetrating wound with large skin defect. Regarding the operative technique, a meticulous debridement and the appropriate flap area were required, and the blood vessel of the flap should not be compressed by the surrounding tissues. The advantages of this operation provided a simple method to handle, high rate of survival, and one-staged operation. PMID- 9867988 TI - [Repair of bone defect of distal end of radius and its functional restoration]. AB - The treatment of the bone defect of the distal part of the radius included repair of the bone defect and restoration of the function of the wrist joint. Since 1979, three operative methods were used to treat 13 cases, and they were grafting of the vascularized fibula by anastomrsis fibular vessels, grafting of upper part of fibular with lateral inferior genicular artery and grafting vascularized scapula flap. Follow-up had been carried out from 1 to 10 years. The result was satisfactory. The discussion included the repair of the defect of the middle or distal part of the radius, the operative methods, main attentions and indications. It was considered that it should be based on the length of bone defect while the operative method was considered. PMID- 9867989 TI - [Choice of treatment for fracture of distal tibia with ankle joint involvement]. AB - It is difficult to treat the intra-articular fracture of distal tibia or Pilion's fracture. From 1987 to 1995, 43 cases of Pilon's fracture were admitted and treated with different methods. After treatment, they had been followed up for 1 to 8 years. According to Riiedi's Classification, there were type I 12 cases, type II 21 cases and type III 10 cases. The methods used on these patients included manual reduction and plaster of paris immobilization, calcaneous tubercle traction and plaster of paris immobilization, open reduction and internal fixation with Kirschner wires, and open reduction and internal fixation with AO plate. The outcome was evaluated according to Ovadia's criteria, for type I fracture, 8 cases were treated with conservative method with a satisfactory rate of 79.17% and 4 cases were treated with operation with a satisfactory rate of 91.67%; for type II, conservative method for 12 cases and the satisfactory rate was 33.33%, and operation for 9 cases with a satisfactory rate of 70.37%; for type III, conservative method for 2 cases, with poor result, and operation for 8 cases with satisfactory rate of 79.17%. The indication for conservative treatment was type I fracture. It showed that for Pilon's fracture, the outcome of open reduction and internal fixation was superior to that of the conservative treatment, especially in those having internal fixation with AO plate. PMID- 9867990 TI - [Repair and functional reconstruction of war wounds of limbs]. AB - From 1979 to 1994, reparative and reconstructive surgery were used to repair the war injuries of skins, bones, blood vessels and nerves of the limbs in 800 cases. A systematic clinical study was carried out. Many new operative methods were used and the results of treatment were good. Innovations and modifications were made in technique. In 120 cases of war injuries having soft tissues defects including skin and muscles, various tissue transplantations were used with the hope to accomplish one-staged repair of the defect and reconstruction of motor function of muscle. To those infections of bone and joint in war injuries, following early eradication of infected focus, transplantation of musculo-cutaneous flap or omental graft was immediately carried out with the aim to obtain primary healing of the wound. In the treatment of bone defects from war wounds with loss of skin and muscles, the vascularized skeleto-cutaneous graft was used. In the treatment of 150 cases of injury of peripheral nerve from forearms, the result of good to fair rated 68.8 percent for upper extremity and that for lower extremity, it was 62.2 percent. Following the early repair of 500 cases of injury of peripheral blood vessels, the patency rate of the blood vessel was 90 percent. The result following by pass vascular graft in the treatment of forearms injury of blood vessels even with very poor local condition was still very successful. PMID- 9867991 TI - [Study on culture and osteogenic potential of stromal cell of bone marrow in vitro]. AB - The osteogenc potential of bone marrow has been proved by experiment. To investigate more in details, bone marrow was obtained from the trochanteric region of femur of New-Zealand rabbit in 4 to 8 weeks old. After being cultured in vitro for one week, the hematopoietic component of the bone marrow had disappeared, thus the stromal cells were obtained. Then the stromal cells were subcultured in cultural fluid containing dexamethasone (10(-8) mol/L) and natrium glycerophosphate (10 mmol/L). Under the phase-contrast microscope, it was found that being cultured for 15 days. The stromal cells were lined up in one layer and late the secretion activity was increased and gradually transformed into multilayer structure and was congregated into diffused opaque clusters in twenty days. During culture, the cells were examined by tetracycline fluorescence label, histochemistry stains, transmission electron microscopy, scanning electron microscopy and energy dispersive X-ray microanalysis. The results showed that the morphological and biological characteristics of the cultured stromal cells derived from the bone marrow were similiar to those of osteoblasts and could synthesized mineralized new bone tissue in vitro. PMID- 9867992 TI - [Influence of reperfusion following ischemia on microvessels and microcirculation of skeletal muscle]. AB - In order to study the influence of reperfusion following ischemia on microvesseles and microcirculation of skeletal muscle, unilateral hindlimbs of 16 rabbits were subjected to normothermic ischemia for 2 and 5 hours by tourniquet. After release of the tourniquet, microcirculation of the peritenon on dorsum of the foot was observed for 1 hours by intravital microscope. At 1 hour and 72 hours following reperfusion, the anterior tibia muscle biopsiy were taken and the specimens were subjected to light and electron microscopic examinations. It was found that after release of the tourniquet, in the limbs undergone 2 hours ischemia, there was immediate and well distributed reflow in the microvesseles of peritenon though a few aggregates of red cells and increase in the number of adherent leukocytes occured in some venules, and the microvesseles of the skeletal muscle only showed signs of minimal injury, the muscle fibers could survive in the limbs undergone 5 hours of ischemia, however, there was serious disturbance of microcirculation in the peritenon, which was characterized by "no reflow" in most area and there was significant increase in the number of leukocytes adherent to venular endothelium, and the microvesseles of the skeletal muscle showed signs of severe injury, including remarkable swelling of the endothelial cell, disruption of the basement membrane and interstitial edema, and finally, most of the muscle fibers had necrosis occured. The results demonstrated that reperfusion following ischimia might result in microvascular injury and microcirculation disorder in the ischemic area. The degree of the injury and disorder depended on the duration of ischemic period, and was an important factor which determined the fate of the parenchymal cell. PMID- 9867993 TI - [Experimental study of the mechanism on blood supply to delayed separated skin flap by ultrasonic Doppler]. AB - In order to investigate the mechanism of blood supply to the delayed separated skin flap and the time and criteria for its transfer, 5 small-sized Banna pigs were selected to produce 14 skin flaps. In the experimental group the skin flap on one side was made from the middle of the back having an extra-thin steel sheet intervened between the flap and the soft tissue of the back whereas the flap on the opposite side without a steel sheet intervened was served as the self control. The skin flaps were examined 3,7,10 and 20 days after the operation and 3,7 and 10 days after transfer, respectively, by (1) gross observation; (2) ultrasonic Doppler; (3) superficial skin temperature measurement and; (4) histomorphological examination. In the experimental group the survival rate of the flap was 100 percent whereas in the control group all of the flaps had necrosis from 30 to 50 percent after the flap being transfered. In the experimental group, the echo sound from the arterial blood flow from ultrasonic Doppler was heard at the pedicle 7 days after the operation, and as time elapsed, the echo sound spread distally, whereas in the controls no echo sound could be heard over the skin flap. There was significant difference statistically between the experimental and control groups in the temperature of the flaps while the flaps were being transferred (P < 0.01). The external diameters of the blood vessels in the central area of the skin flap were larger in the experimental group, 0.8 to 1.2 mm in comparison to 0.4 mm. PMID- 9867994 TI - [Absorbable materials and their uses in surgery]. PMID- 9867995 TI - [Experimental study on the collateral sprouting after end-to-side anastomosis of nerve trunk]. AB - In order to observe the collateral sprouting capacity of the nerve trunk after end-to-side anastomosis, a window was made on the epineurim of the donor nerve through which the result of the end-to-end anastomosis of nerves could be compared. Sixteen SD rats were chosen and divided into 4 groups randomly. Group 1, the peroneal nerve was severed, the epineurium of the tibial nerve was fenestrated and sutured the peroneal stump with the tibial nerve by end-to-side neurorrhaphy. Group 2, operative procedures were almost the same as that of Group 1, but no fenestration on the epineurium was done. Group 3, the distal peroneal nerve stump was sutured in paralell with the tibial nerve without fenestration on the tibial nerve and Group 4, severed the peroneal nerve and sutured the stumps by end-to-end anastomosis immediately. The peroneal function index(PFI) and acetylcholine transferase (ChAT) activity were assessed and the histological examination was performed in all rats. The results showed: between group 1 and 2, there was no difference in PFI and ChAT activity (P > 0.05). The existence of collateral sprouting in all groups was proved by histological examination. Even in Group 3, there were plenty of nerve fibers turned into tiny myelined nerve fibers through collateral sprouting. But the ChAT activity in Group 1 was only two thirds of that in Group 4. It was suggested that the epineurial sheath did not influence the collateral sprouting of the nerve. PMID- 9867996 TI - [Experimental study on prevention and treatment of neuroma by implanting nerve stump into muscle]. AB - Prevention and treatment of traumatic neuroma by implanting the proximal neural stump into the muscle were studied. Sixteen SD rats were used for the experimental study. The proximal stump of the left sciatic nerve was implanted into the nearby muscle as the experiment side, whereas the proximal stump of the right sciatic nerve was left untreated as the control side. The results were assessed with histological and electrophysiological methods. The experiment demonstrated that neuroma was formed in the control side one month postoperatively, whereas in the experimental side the nerve fibers were dispersed among the muscle fibers and no definite neuroma was formed. Implantation of neural stump into muscle could prevent and treat traumatic neuroma. PMID- 9867997 TI - [A comparative study of laryngeal reinnervation from the ansa cervicalis]. AB - A comparative study of four methods of laryngeal muscle reinnervation in dogs is presented. Twenty-eight cases were divided into four groups to undergo main branch and branch of ansa cervicalis nerve anastomosis, and nerves implantation an neuromuscular pedicles transfer respectively for restoration of vocal cord adduction on left sides. The results showed that the four procedures seemed to induce effective reinnervation of adductor muscles. But the main branch of ansa cervicalis nerve suture was superior to the other methods among which little difference was noted in the functional recovery, electrophysiological activity and muscle strength. It demonstrated that main branch of ansa nerve suture was the best procedure for treatment of unilateral vocal cord paralysis among the four methods. PMID- 9867998 TI - [Recovery of motor function of spinal cord with embryonic spinal cord graft in adult rat]. AB - The capacity of embryonic spinal cord tissue in the repair of injured structure of spinal cord has been noted for years. In order to investigate the embryonic spinal cord graft in the repair of motor function of injured spinal cord, the embryonic spinal cord tissue was transplanted to the hemisection cavity in spinal cord in adult rat. One hundred adult Wistar Rats were used to simulate the hemisectional injury of spinal cord by drilling 2-3 mm cavity in lumbar enlargement. Sixty rats were treated with rat embryonic spinal cord tissue grafting while the other forty were chosen as control. The outcome was evaluated according the combined behavioural score (CBS) and motor evoked potential (MEP) in the 1, 2, 4 and 12 weeks. The grafting group was superior to the control as assessed by CBS (P < 0.05), especially within 4 weeks. (P < 0.01). The restoration of the latent peak of early wave(P1, N1) was better in the grafting group, too. This suggested that embryonic spinal cord graft could improve the recovery of motor function of injured spinal cord in adult rat. The effect of the embryonic spinal cord tissue graft might be concerned with its secretion of several kinds of neurotrophic factors, nerve growth factor, nerve transmitted factor, or adjustment of hormone. PMID- 9867999 TI - [Anatomical study and clinical application of transfer of pronator quadratus branch of anterior interosseous nerve in the repair of thenar branch of median nerve and deep branch of ulnar nerve]. AB - In order to obtain the anatomical basis of transferring the anterior interosseous nerve to repair the injury of thenar or ulnar nerve at the wrist level, ten fresh cadaveric forearms were dissected. The pronator quadratus branch of the median nerve was (1.5 +/- 0.4) mm in diameter with (866 +/- 144) nerve fibers. The recurrent branch of median nerve was (1.7 +/- 0.3) mm in diameter with (1,120 +/- 97) nerve fibers. The deep branch of ulnar nerve was (2.1 +/- 0.4) mm in diameter with (1,318 +/- 120) nerve fibers. To repair the thenar recurrent branch, nerve graft should be used to bridge between the distal end of the anterior interosseous nerve and the origination of the recurrent branch. The deep branch of the ulnar nerve could be repaired by direct suture after being dissected proximally. In patients, the technique was applied to repair 17 cases of injury of thenar branch and 3 cases of injury of deep branch of ulnar nerve. Among them, seventeen cases were followed up from 2 to 7 years. On electromyogram it was normal in 10. The myodynamia was restored in different degree: M2 in 2, M3 in 5, M4 in 7, M5 in 3. The procedure of the operation and the matters needed attention during the operation were discussed. PMID- 9868000 TI - [Preliminary investigation of treatment of ulnar nerve defect by end-to-side neurorrhaphy]. AB - In the repair of the defect of peripheral nerve, it was necessary to find an operative method with excellent therapeutic effect but simple technique. Based on the experimental study, one case of old injury of the ulnar nerve was treated by end-to-side neurorraphy with the intact median nerve. In this case the nerve defect was over 3 cm and unable to be sutured directly. The patient was followed up for fourteen months after the operation. The recovery of the sensation and the myodynamia was evaluated. The results showed that: the sensation and the motor function innervated by ulnar nerve were recovered. The function of the hand was almost recovered to be normal. It was proved that the end-to-side neurorraphy between the distal stump with the intact median nerve to repair the defect of the ulnar nerve was a new operative procedure for nerve repair. Clinically it had good effect with little operative difficulty. This would give a bright prospect to repair of peripheral nerve defect in the future. PMID- 9868001 TI - [Clinical application of implantation of vascular bundle into silicone tube to bridge the peripheral nerve defect]. AB - In order to improve the therapeutic effect of non-neural tissue in bridging the peripheral nerve defect and increase the blood supply of the implant, the silicone tube was chosen to bridge the gap, and the vessel bundle was inserted into the tube. The procedures were performed as following: resected the pseudoneuroma and enveloped the proximal and the distal ends in a silicone tube, and then sutured the epineurium and the tube wall with 7/0 stitch. In patients, eleven cases with fifteen nerves were treated, including seven median nerves, five ulnar nerves and three radial nerves. The lengths of the nerve defects were within 3 cm in 13 nerves and 3 cm-5 cm in 2 nerves. They were followed up from one to five years and the result was excellent (M4S4) in 8 nerves, good (M3S3) in 3 and poor (M1S1) in 2. It was discussed that the indication for the procedure should be included: the nerve defect could not be sutured directly, and the patient would not agree to use his own nerve for graft. PMID- 9868002 TI - [Peripheral nerve injury as a complication from orthopedic operation]. AB - Nerve injury following operation is one of the main causes of the iatrogenic peripheral nerve injury. In order to learn lessons from these cases, one hundred and seven cases of peripheral nerve injury complicated with the orthopedic operations were analyzed. Forty-four cases were cutting injury during operation, made up 41% of all cases and 27 cases were stretch and compression injury, made up 25%. The involved nerves included 41 radial nerves and 24 common peroneal nerves, composing 60.7% of all nerve injury. The operations responsible were mainly the bone and joint operations, which made up 81%. The cause, prophylaxis, diagnosis and treatment were discussed. The rich appropriate knowledge of anatomy and responsibility of the surgeon were emphasized in order to prevent the occurrence of complication. Once the injury was suspected, diagnosis should be made promptly and effective treatment should be performed in time. PMID- 9868003 TI - [Experiment and clinical application of squeezing technique in venous crisis after replantation of finger]. AB - In search of a new method for treating venous crisis after replantation of the finger, the effect of instantaneous, quick and forceful squeezing on the belly of replanted finger (the so-called SQUEEZINGG TECHNIQUE) was studied. In the animal experiment, 20 SD rats were used. The femoral veins and arteries were separated in order to measure the venous pressure. The result showed that the venous pressure was (8.33 +/- 1.29) x 10(-2) kPa in normal condition, (20.61 +/- 2.34) x 10(-2) kPa in the condition simulating venous crisis and (73.9 +/- 5.74) x 10(-2) kPa on carrying out the squeezing technique. The second part of the experiment was performed on ten human fingers which were amputated because of trauma or other diseases. One of the arteriae digitales palmares propriae and two of its accompanied veins were disected, and all other blood vessels were ligated. The artery was irrigated with normal saline at a pressure of 12 kPa. When both veins were left open, the venous pressure was (1.32 +/- 0.17) kPa; (4.29 +/- 0.49) kPa, when both were ligated; and (16.88 +/- 5.25) kPa when the squeezing technique was applied at the time venous crisis developed. From May 1991 to May 1996, this method was used for 43 times in 25 cases (35 fingers) with venous crisis after replantation. It was successful for 39 times in 21 cases (30 fingers). It was very important that the performer should use the thumb and the index finger to squeeze just the belly of the involved finger, and that every squeeze should be forceful, quick and instantaneous. It was suggested that if this method was properly performed it could improve the survival rate of replanted fingers. The mechanism of squeezing technique in the treatment of venous crisis following replantation of finger was proposed. PMID- 9868004 TI - [Thumb and finger reconstruction with the pedal digit transplantation: 541 cases report]. AB - Thumb and finger reconstruction by the method of pedal digit transplantation had been successfully performed in 541 casee from 1977 to 1996, which contained 404 cases of thumbs and 78 cases of fingers. The thumb reconstruction was mainly the simple transplantation of distal phalanx (42 cases) and the compound transplantation of hallucal nail-cutaneous flap with iliac bone segment (16 cases) for the defect of thumbs in degree 1 and 2. The combined transplantation of hallucal nail-cutaneous flap with the joint and tendons of the second toe (34 cases) and the transplantation of the distal part of the second toe (182 cases) for the defect of degree 3 and 4. The combined transplantation of the second pedal digit with its metatarsalphalangeal joint (189 caese) for the defect in degree 5 and 6. The finger reconstruction was performed by anastomosis of the arteries of the digit with those of the fingers for 29 cases with the defect in degree 2 and 3, 60 cases with the defect in degree 4 and 5, and 17 cases with the defect in degree 6. One-hundred and four cases of versels vasiation were found in this group (19 cases with the pedal dorsal artery, 13 cases with the greater saphenous vein and 72 cases with the first dorsal metatarsal artery). The main point of the operation and the treatment of the vessel variations were discussed. PMID- 9868005 TI - [Thin skin flap in the repair of defect of soft tissue of hand]. AB - To meet the special need of function and cosmetics, thin skin flap was used in repair of skin defect of hand. It was reported that 37 cases with skin defect of hand had been treated since 1992. Eight kinds of flaps were used, including sub abdominal flap (in 8 cases), paraumbilical flap (in 8 cases), intercostal flap (in 5 cases), subclavicular flap (in 3 cases), thoracoacromial flap (in 3 cases), trigonum clavopectoral flap (in 4 cases), thenal flap (in 2 cases) and pedicled flap carrying superficial iliac circumflex artery (in 4 cases). All of defects were treated by one-stage operation. The size of the flaps used ranged from the minimum of 1.2 cm x 2.5 cm to the maximum of 10 cm x 16 cm. All the flaps were survived. The function and appearance of the hands were satisfactory. It was noted that thin skin flap had developed from conventional pedicled flap and subcorium vascular-net flap, but it was easy to perform and had a high rate of success. The principle in choosing the flap was also discussed. It was suggested that more attention should be concentrated to solve the problems encountered in the use of thin skin flap in the repair of skin defect. PMID- 9868006 TI - [Three kinds of distally based fascial flap containing deep fascial vessel network on the posterior calf]. AB - Considering the abundant vascular anastomotic networks in the deep fascia of the posterior calf, three kinds of distally based facial flap containing deep fascial vascular network were applied clinically. They were: 1. posterolateral distally based island fascial flap which could be used to repair the skin defect of heel, dorsum of foot and lateral-distal part of leg; 2. posteromedial distally based island fascial flap which could be used to repair the skin defect of heel, medial malleolus and medial-distal part of leg and 3. posterolateral malleolar distally based fascial flap which could be used to repair the skin defect of heel and lateral malleolus. Eighteen cases with soft tissue defects around the distal calf were treated, the area of skin defect ranged from 4 cm x 3 cm to 13 cm x 6 cm. All the flaps were survived completely after operation with an average of follow up for 15 months (ranged from 6 months to 2 years). So the advantages of these flaps were as follows: the blood supply was reliable, preparation of the flap was easy and the major arteries of the calf needed not be sacrificed; the flap had a long and rotatable pedicle so that they would basically satisfy the need to repair skin defect of lower leg, dorsum of foot, heel and malleolus and the resistance of the flap to pressure and wear was better. However, the injury to the superficial sural nerve was the shortcoming. PMID- 9868007 TI - [The application of distally based fascial pedicled island flap to rapair wound of the extremities]. AB - In order to preserve the major vessels of the extremities in the repair and reconstruction of wounds of the extremities, the distally based fascial pedicled island flap was applied clinically. Its axis and rotatary point were designed along orientation of the major arteries, and the blood supply was from the abundant vascular networks in the deep fascia. Twenty-two cases with exposure of tendon and bone including 10 upper limbs and 12 lower limbs were treated. The flap area of forearm ranged from 7 cm x 8 cm to 12 cm x 9 cm and the ratio of the length to width of the pedicle was 1: 1-2. The flap area of the calf ranged from 10 cm x 6 cm to 16.5 cm x 12 cm and the ratio of the length to width of the pedicle was 2:1. The rotatary angle was 130 degrees-170 degrees. After operation, 18 flaps were survived completely, 2 cases had partial necrosis on the margin, 2 failures received cross-leg flap in the second operation. The patients were followed up with an average of 13.5 months (ranged from 3 months to 2 years). The conclusions were as follows: 1. the blood supply of this type of flap was reliable and the major arteries of the extremities needed not to be sacrificed; 2. the preparation of the flap was easy and the survival rate was satisfactory; 3. the shortcomings of this flap were unsightly incision scar and the limited size of flap and; 4. during the operation, the compression of the pedicle must be avoided. PMID- 9868008 TI - [Use of fibrin glue in the prevention of secondary anastomotic stenosis from repair and reconstruction of the injury of the bile duct]. AB - The secondary anastomotic stenosis is often occured from the repair and reconstructive operation of the injured bile duct. It is difficult to treat and the outcome is serious. In order to prevent this complication, the fibrin glue instead of traditional suturing technique combined with inner support was used. Fifty-four hybrid dogs were divided into 3 groups. Group A received Roux-en-y choledochojejunostomy with fibrin glue; group B received Roux-en-y choledochojejunostomy, with a fibrin glue combined support left permanently in the bile duct and group C received Roux-en-y choledocholejejunostomy with fibrin glue combined a support left temporarily in the bile duct. The amount of collagen in the scar was measured at 3/4, 3, 6, 9, 12 months respectively after operation. The results showed: 1. the mature period of scar was shortened from 12 months to 9 months when fibrin glue instead of suture was used in choledochojejunostomy; 2. the mature period of scar was further shortened from 9 months to 6 months when fibrin glue combined with inner support instead of fibrin glue was used in choledochojejunostomy. The conclusions were as follows: 1. fibrin glue could facilitate the healing of wound by inhibiting the formation of scar and accelerrate the maturation of scar; 2. when the inner support was used with fibrin glue in the operation, the mature period of scar could be further shortened; 3. the mechanism of action of the fibrin glue included minimizing the injury, avoiding foreign-body reaction, modifying organization of hematoma, preventing formation of biliary fistular and enhancing intergration and cross linkage of collagen. PMID- 9868009 TI - [Current status and trends of guided tissue regeneration materials]. PMID- 9868010 TI - [The expression of IGF-1 and its receptor in cultured tendon cell]. AB - In order to study the expression change of insulin-like growth factor-1 (IGF-1) and its receptor genes in different generations of tendon cell in culture, Dig labeled synthesized oligonucleotide probes were used to detect the mRNA expression in primary, 6th and 13th generation of tendon cell. The results showed that IGF-1 receptor mRNA was expressed in all of the 3 above generation tendon cells. IGF-1 mRNA was expressed only in primary and 6th generation cells. Tendon cell of 13th generation did not express IGF-1 mRNA. It might suggest that the absence of IGF-1 mRNA expression be one of the causes which led to the decrease of reproductive ability of 13th generation tendon cell. PMID- 9868011 TI - [Analysis of pure titanium implants-bone interface by TOF-SIMS]. AB - In order to study the Titanium-bone interaction and integration mechanism, the titanium implant was implanted in the tibia of 9 Newzealand rabbits, and the Ti bone interface performed for 1, 3, and 6 months were examined and analyzed by fluorescence microscope and advanced TOF-SIMS techniques. The results showed that Ti-bone tissue was integrated closely in a very reactive manner. Both physical and chemical integration occurred in the Ti-bone interface. The Ti-bone could diffuse into the bone tissue though the diffusion was very limited. It was up to 100 microns in depth during the early period. The diffusion density was high, and later in a smooth distribution. Furthermore, while Ti+ diffused into the bone tissue, other elements such as Ca+, OH-, O-, etc, could also diffuse into titanium in exchange. The growth pattern around the bone tissue was in two fashions, one was implantefugal and the other was implantopetal. In this study, based on the ionic distribution, osmosis and impurity elements distribution, the Ti-bone integration mechanism was discussed at molecular and atomic level. PMID- 9868012 TI - [The effect of vascular endothelial cell growth factor on survival of skin flap in rats]. AB - In order to study the effect of vascular endothelial cell growth factor (VEGF) on the survival of skin flap 30 SD rats were used. A randomized flap measuring 7.5 cm x 3.0 cm was created on the back of each SD rat. The treatment group (n = 10) received VEGF 40 ng/flap by subcutaneous injection with microinjector during and 24 hours after operation. The control groups received heparin 16 U/flap (n = 10) or normal saline 800 microliters/flap (n = 10). After operation, on the 3rd and 11th day, the survival rate of the skin flaps and the dermovascular density of each flap were investigated by histological and histo-morphometrical examination. The results showed that there was no significant difference in the survival rate between the treatment group and the controls on the 3rd day after operation, while on the 11th day, there was a significant difference between them, and the survival rate was much higher in the treatment group. Besides, dermovascular density was much more increased in the treatment group than that in the controls, especially in the distal 1/3 of the flap (P < 0.02). The conclusion was that VEGF could accelerate the vascularization of the flap and enhance the survival rate of the flap. PMID- 9868013 TI - [Experimental study of skin expansion of the grafted free vascularized skin flap]. AB - By using retro-auricular skin after skin expansion to reconstruct auricle, often times, the retro-auricular skin could not be expanded because of certain reasons such as having burn scar. For this purpose, a new operative technique was designed. The experiment consisted of using 15 rabbits, on one side of the back, a free vascularized skin flap was obtained. Following the anastomosis of the vessels of the flap with that of the back the flap was sutured back in situ. Two weeks after the removal of the sutures, the grafted skin flap was expanded as usual. The routine skin expansion was carried out on the contralateral side as control. The volume of expansion, change of subcutaneous blood vessels, complications from operation, and histological observation were used to compare the results of skin expansion of the two sides. The result showed that there was no significant difference between the types of skin expansion. It was concluded the grafted free vascularized skin flap had no adverse effect on skin expansion. PMID- 9868014 TI - [Repair of long segment bone defect of femur by free juxtaposed bilateral fibulae autograft]. AB - There were several methods, such as free single and folded fibulae autograft, composed tissue autograft, however, it is still very difficult to repair long segment bone defect. In December 1995, we used free juxtaposed bilateral fibulae autograft to repair an 8 cm of femoral bone defect in a 4 years old child in success. The key procedure is to strip a portion of the neighboring periosteal sleeve of juxtaposed fibulae to make bare of the opposite sides of the bone shafts, suture the opposite periosteal sleeves, keep the nutrient arteries, and reconstruct the blood circulation of both fibular by anastomosis of the distal ends of one fibular artery and vein to the proximal ends of the other fibular artery and vein, and anastomosis of the proximal ends of the fibular artery and vein to lateral circumflex artery and vein. After 22 months follow up, the two shafts of juxtaposed fibulae fused into one new bone shaft. The diameter of the new bone shaft was nearly the same as the diameter of the femur. There was only one medullary cavity, and it connected to the medullary cavity of femur. This method also cold be used to repair other long segment bone defect. PMID- 9868015 TI - [The protective role of heat shock gene expression on hydrogen peroxide induced pulmonary alveolar macrophages]. AB - The aim of this study is to determine whether heat-induced heat shock gene expression in rat pulmonary alveolar macrophages (PAMs) would protect PAMs against hydrogen peroxide (H2O2)-mediated cell killing. In response to sublethal heat shock at 42 degrees C for 2 h, the cells synthesized heat shock protein 70 kD (HSP70) and other different molecular weight heat shock proteins, which were detected with gel electrophoresis after [35S]-methionin labeling cellular protein and Western blotting analysis in PAMs. Northern blot analysis showed the induction of HSP70 mRNA with heat shock treatment. Futhermore, with heat shock, there was a significant increment of survival cells after H2O2 (1,2,3 mmol.L-1, 45 min) exposure. This increase was blocked both by Cycloheximide and by Actinomycin D, and was associated with inhibition of synthesis of HSP70 protein and transcription of HSP70 mRNA. These results strongly suggest that expression of heat shock gene (especially HSP70 gene) plays an important role in the intracelluar mechanism of cytoprotection against H2O2 in rat PAMs. PMID- 9868017 TI - [Effect of ischemic preconditioning on adenine nucleotide levels of graft lung from canine donor]. AB - Twelve canine left lung allotransplantation were performed. In the ischemic preconditioning group (Group IP, n = 6), left donor lung was preconditioned with 10 min ischemia followed by 15 min reperfusion using the occuluding left hilum before resection and cold perfusion. The control group (Group C, n = 6) underwent the same treatment but without ischemic preconditioning. Adenine nucleotides (ATP, ADP, AMP) of the donor lung tissue were measured using rHPLC after 2 hr of resection and cold perfusion with Euro-Collins solution (ECS). The results showed that contents of ATP and total adenine nucleotides (TAN) were much higher in Group IP than in Group C (322.9 +/- 61.2, 942.9 +/- 134.5 and 200.0 +/- 50.0, 668.4 +/- 59.6 mumol.g-1 respectively, P < 0.05). Histologic examination of the donor lung in Group IP showed less damage than in Group C after 2 hr of transplantation. The results suggest that IP combined with cold ECS perfusion can reduce the energy metabolism in canine donor lung. PMID- 9868016 TI - [Preliminary application of PCR to curative effect assessment of chemotherapy against experimental acute infection of Toxoplasma gondii in mice]. AB - Treatment with pyrimethamine and sulfamethoxazolum combination administered via intragastric route, was started 2 hours after each mouse received an intraperitoneal injection of a lethal inoculum of 5 x 10(3) tachyzoites of RH strain toxoplasma gondii and the therapy was lasted for 52 days. Combined therapy of the two drugs resulted in 83.3% survival rate in mice. PCR and the conventional mouse inoculation (MI) were compared for their validity as an curative effect assessing method. The positive rates of PCR and MI were 63.64% and 18.18%, respectively. The results suggest that PCR can be used in studies involving drug screening and curative effect assessment of anti-Toxoplasma chemotherapy in vivo. In addition, the results indicated that longer course of treatment against toxoplasmosis is required in clinical practice. PMID- 9868018 TI - [Changes of NMDA receptor gene expression in infection brain injury induced by Bordetella pertussis in rats]. AB - N-methyl-D-aspartate receptors (NMDAR) in cerebral cortexes of rat brain injury induced by Bordetella pertussis (Bp) were measured by methods of Northern blot analysis and [3H] MK-801 radioligand-receptor binding to study changes and effects of NMDAR1 of gene expression in the infection brain injury. The affinity (Kd) of NMDAR in Bp group was significantly decreased [30.5 +/- 3.0 nmol.L-1 (Bp), 43.1 +/- 4.2 nmol.L-1 (NS), P < 0.05], but expression of NMDAR1 mRNA and binding site density (Bmax) of NMDAR observed no significant differences between the experimental and the control group (P > 0.05), respectively. The results suggest that NMDAR1 was activated with high affinity functionally, but no changes in binding site density and transcriptional expression of NMDAR1 occured in the infection brain injury. PMID- 9868019 TI - [Effect of Ganoderma japonicum (Fr.) Lloyd mixture on experimental thrombosis]. AB - The therapeutic effect of Ganoderma japonicum(Fr.) Lloyd mixture on thrombosis and its mechanism were studied. The results showed that Ganoderma japonicum(Fr.) Lloyd mixture inhibited thrombus formation in vitro and in vivo, the thrombus weight and length formed in the rabbit common carotid artery and external jugular vein were significantly decreased in the experimental group compared with the control (P < 0.01). The results suggest that Ganoderma japonicum(Fr.) Lloyed mixture has anti-thrombotic effect, blood coagulation and platelet activation were inhibited, and the ability of vascular endothelial cells against the process of thrombosis was enhanced. PMID- 9868020 TI - [The effect of wuto granules on the levels of excitatory amino acids in extracellullar fluid of rat hippocampus following cerebral ischemia]. AB - We observed the changes of excitatory amino acids (EAA) in extracellular fluid (ECLF) in rat hippocampus in case of cerebral ischemia after giving Wuto Granule compring the test groups with the control groups by microdialysis method. Results showed that there was no effect on EAA in extrcellular fluid by the drug before cerebral ischemia, but there existed staistical singnifcant changes of EAA in extracellular fluid one hour after cerebral ischemia comparing the test groups Glu and Asp with the control groups (P < 0.01). We found that there is a decrease of release of release of EAA in ECLF in case of ischemia by Wuto Granule. It could be deduced that Wuto Granule can penetrate the brain-blood barrier and this can protect the brain incse of ischemia. The application of intracerebral micrdialysis has provided a new method for the research work of pharmacology. PMID- 9868021 TI - [The protective effect of captopril on the ischemic myocardium in dogs through coronary sinus retroperfusion]. AB - The protective effect of coronary sinus retroperfusion of Captopril on the ischemic myocardium was observed in dogs with acute myocardial infarction (AMI). The results showed that the infarction size and the level of coronary sinus plasma endothelin (ET) and manodialdenyde (MDA) were smaller and lower when Captopril was administered by coronary sinus retroperfusion than that by systemic intravenous injection. These results suggest that (1) Captopril can be distributed adequately in the local ischemic myocardial zones when administered by coronary sinus retroperfusion in the presence of coronary artery occlusion, (2) Captopril can more effectively protect ischemic myocardium by inhibition of the ET release and against the oxygen free radicals in ischemic myocardial area when used by coronary sinus retroperfusion. PMID- 9868022 TI - [An experimental study of bioactive glass ceramics as orbital implants]. AB - One eyeball of experimental rabbits was enucleated in routine procedure, and then implanted the biocative glass ceramic (BGC). No rejection was observed during the postoperation period. After three months of implantation, they were examined with ultrasound, and a venous-flow-like spectra was detected in the ceramic implant. After six months of implantation, the implans were examined histologically; the ingrowth fibrovascular tissue filled the 90% gaps of the implant. The results suggest that the bioactive glass ceramic implants made in China possess acceptable histocompatibility. PMID- 9868023 TI - [The contents of plasma norepinephrine and epinephrine in patients with five types of Gan syndromes in TCM and its significance for diagnosis]. AB - Plasma norepinephrine (NE) epinephrine (E) concentrations in patients with five types of Gan syndromes and relative syndromes were determined. The healthy individuals served as control. The contents of NE and E in Ganyang Huafeng Zheng (5975.8 +/- 305.6 pmol.L-1, 2227.7 +/- 12.57 pmol.L-1) were significantly increased; Ganxue Xu Zheng (1100.4 +/- 105.5, 451.2 +/- 70.9 pmol.L-1) were decreased, and lower than the normal controls. NE and E contents in both Ganyang Shangkang Zheng (3962.6 +/- 354.4, 1225.6 +/- 163.5 pmol.L-1) and Ganhuo Shangyan Zheng (3398.1 +/- 303.8, 1134.4 +/- 146.9 pmol.L-1) were increased remarkably, but there was no difference between them. Plasma NE, E concentrations can be considered as one of the comprehensive laboratory parameters in diagnosis of the four above mentioned Gan syndromes. PMID- 9868024 TI - [Diagnostic value of the heart-reactive autoantibodies for viral myocarditis in children]. AB - The serum heart-reactive autoantibodies (HRAA) of 103 children with viral myocarditis (VMC) were detected on normal myocardial tissues of human embryo with the use of indirect immunofluorescence assay and were compared with the result of 55 children with other diseases (COD) and of 67 healthy children (HC). The results showed that the positive rate and the geometric mean titre (GMT) of HRAA in VMC group were obviously higher than that in the other groups (P < 0.01). If we took the HRAA upper limit (1:9.5) of 99% normal value range in HC group as the positive criterion, the diagnostic sensitivity of HRAA to child VMC would be 71.84%, the specificity of differential diagnosis of VMC from COD and HC would be 76.36% and 91.04% respectively. The coincidental rates of diagnosing VMC from COD and HC were 73.41% and 79.41%. Four patterns of immunofluoresent localization and shape could be distinguished as scatter, spot, membrane and cord. These indicate that HRAA is important in diagnosing child VMC. PMID- 9868025 TI - [Color Doppler flow imaging study on the changes of collateral circulation between portal-superior vena cava and azygos vein before and after endoscopic ligation of the esophageal varix]. AB - Color Doppler flow imaging (CDFI) was performed in 35 patients with portal hypertension and esophageal varix. Thirty subjects were served as control. After esophageal variceal ligation, CDFI observed that the portal-superior vena cava collateral veins were partialy blocked. The esophageal varices disappeared. The esophageal wall thinned. Diameter of left gastric vein enlarged and blood flow velocity decreased. Azygos diameter was reduced and blood flow velocity was decreased. Although diameter of portal vein was not changed, the blood flow velocity slightly increased. The results suggest that ligation treatment can have the tendency to increase blood flow of liver and stomach, which might aggravate gastric mucosal lesion. PMID- 9868026 TI - [The relationship between expression of basement membrane in squamous cell carcinoma of oral cavity and cervical lymph node metastasis]. AB - Fifty seven cases of oral squamous cell carcinoma (SCC) were studied by immunohistochemical ABC method using type IV collagen and laminin antibody to investigate the relationship between expression and distribution of basement membrane in oral SCC and clinicopathologic characteristics and cervical lymph node metastasis. The distribution of basement membrane of oral SCC was discontinuous and in some cases the membranes disappeared. There was highly significant correlation between the staining patterns together with histologic differentiation degrees and cervical lymph node metastasis (P < 0.05). These indicate that the expression of baasement membrane in oral SCC may be a useful parameter for evaluation of tumor histologic differentiation and tumor invasion and metastasis. PMID- 9868027 TI - [Evaluation of electrophysiological diagnosis of concealed accessory pathway (CAP) during transesophageal atrial pacing (TEAP)]. AB - CAP were diagnosed by TEAP using RPE intervals in 55 cases of paroxysmal supraventricular tachycardia (PSVT). There were no obvious CAP manifestations of ECG in those cases, the mean RPE interval during tachycardia was 140 +/- 29 ms, and the lower limit of x +/- 2 s was 82 ms. Among them, the VA intervals were measured by electrocardiophysiological examination in 21 cases. The mean value was 130 +/- 25 ms, and the lower limit of x +/- 2 s was 80 ms. The results suggest that RPE over 82 ms may be an important diagnostic threshold cut off point of atrioventricular reciprocation. PMID- 9868028 TI - [Musical relaxation therapy on the cerebral arteriosclerosis]. AB - The transcranial Doppler and psychological measurement methods were used to evaluate the effect of musical relaxation therapy on the cerebral arteriosclerosis. The results showed that the musical relaxation therapy could improve the cerebral blood flow (physiological), offset the Type A behavior and eliminate the emotional disturbance (psychological), and that its effect was better than that of the control patients. The mechanism of the effect may be related to the 1/f sound wave (physical), abdominal respiration with long expiration and involuntary inspiration (physiological) and the hinting effects of the inducing phrases (psychological). PMID- 9868029 TI - [The plasma levels of endothelin in diabetic retinopathy and their changes after treatment with doxium]. AB - The plasma endothelin (ET) concentrations were determined in 45 diabetes mellitus patients. The changes of ET in 20 diabetic retinopathy patients before and after treatment with doxium and 30 normal controls were observed. The level of plasma endothelins was higher in diabetes mellitus patients than that of the controls (P < 0.05), which was even higher in diabetic retinopathy (P < 0.01). After treatment with doxium, the plasma endothelins level decreased significantly (P < 0.01). PMID- 9868030 TI - [Effects of lacidipine on serum oxygen free radicals and fasting levels of insulin in patients with essential hypertension]. AB - Thirty patients with mild to moderate essential hypertension had received four weeks treatment with Lacidipine and serum lipid peroxide (LPO), superoxide dismutase (SOD) and insulin levels were measured before and after treatment. The results showed: (1) there were decreased in SOD activity and increased Lpo content the patients group (P < 0.01); (2) after treated with Lacidipine, the SOD acitvity increased and Lpo content decreased (P < 0.01); (3) Lacidipine had obvious antihypertensive effect with no significant changes in heart rate; (4) Lacidipine had no significant effects on insulin, blood glucose and lipids. The results suggest that lacidipine is a safe and effective antihypertensive agent and can improve the metabolic balances in free radical system. PMID- 9868031 TI - [Gamma knife treatment of brain metastatic tumors]. AB - Twenty-one cases of brain metastases including 46 tumors treated with Gamma knife (GK) from Sep. 1995 to July 1996 were reported. Among them, 4 patients died within one month, 17 patients with 29 tumor masses were followed up by MRI or CT. The rate of local control was 93%. The total mortality was 19%. The GK treatment associated mortality was 4.7%. Karnofsky Performance Status (KPS) stabilized in 3 cases, improved in 12 cases, regressed in 2 cases. One patients was operated due to the increase of the tumor volume. Four patients were treated with GK again due to the new lesions. The indications of GK treatment of the brain metastases, dose planning, and efficacy were discussed in brief. PMID- 9868032 TI - [Clinical evaluation of polyamine assay in diagnosis of malignant ascites]. AB - Polyamine levels were determined by precolumn derivation RH-HPLC in 68 patients with ascites including 21 malignant ascites, 28 liver cirrhosis and 19 tuberculosis. The total polyamine, especially the contents of cadaverine (CA) and spermidine (SPD) in malignant ascites were significantly higher than that in cirrhosis and tuberculosis. Among them CA levels in malignant ascites and cirrhotic and tubercular ascitic fluid were 2.26 +/- 1.83 nmol.ml-1, 0.75 +/- 0.52 nmol.ml-1 and 0.76 +/- 0.47 nmol.ml-1 respectively, demonstrated the most obvious difference. Using 1.0 nmol.ml-1 as a threshold cut off point to differentiate the malignant ascites from non-malignant ascites, the from non malignant ascites, the sensitivity, specificity and accuracy reached 76. 2%, 83.0% and 80.9% respectively. The results suggest that polymine level in ascites might be regard as one of the cancer-associated markers. PMID- 9868033 TI - [The changes of intraerythrocyte calcium metabolism and their responses to nifedipine in patients with essential hypertension]. AB - The erythrocytic activity membrane ATPases, intraerythrocytic calcium concentration ([Ca2+]i), plasma renin activity (PRA), and angiotensin II (AT-II) in 18 essential hypertensive patients (EH) and matched normal controls were determined. In the EH group, Na(+)-K(+)-ATPase, Ca(2+)-ATPase, PRA were much lower than those in the controls, and [Ca2+]i, AT-II were higher. When blood pressure was become normotensive after a course of taking nifedipine therapy, the forementioned parameter alteration restored to approach the normal range, the changes were significnt. [Ca2+]i was positively correlated with MAP, and negatively correlated with PRA and AT-II. The results suggest that the increase erythrocytic membrane ATPase activity and decrease in renin-angiotensin system activity in nifedipine treatment may be another important antihypertensive mechanism besides accompanied with the rectifying of abnormal cellular calcium metabolism. PMID- 9868034 TI - [Effects of glycyrrhisin on myocardial ischemia/reperfusion injury of rabbits]. AB - The content of MDA and the activity of creatine phosphokinase (CPK) together with the ultrastructural changes in myocardial tissue were analysed to observe the effects of glycyrrhisin on myocardial ischemia/reperfusion injury in rabbits. The results showed that glycyrrhisin could remarkably reduced the level of MDA and the activity of CPK in myocardial tissue (P < 0.01), and mitigated significantly the myocrdial ultrastructural injury in comparison with ischemia/reperfusion group. It was considered that glycyrrhisin exerted obvious protective effects on myocardial injury induced by ischemia/reperfusion. PMID- 9868035 TI - [The intelligence test of the patients with chronic renal insufficiency]. AB - Thirty patients with chronic renal insufficiency (CRF) and twenty-six normal subjects were tested with Wechsler Adult Intelligence Scal (WALS). The verbal intelligence quotients (VIQ), performance intelligence quotients (PIQ), and full intelligence quotients (FIQ) of the patients were lower than those of the normal group (P < 0.01). Among them the mild and moderate intelligence damages were accounted 13.33%. Further analysis indicates that uremic posionous materials damage on the cerebral function was dose dependant. But the disease course was independant with the intelligence damage (P > 0.05). The results suggest that the plasma concentrations of uremia poisonous materials (BuN, Scr, etc.) in CRF patients should be lowered as early as possible in order to keep the normal function of the cerebrum. PMID- 9868036 TI - [Factors analysis of pregnancy success in chronic glomerulus diseases]. AB - Ten cases pregnant chronic glomerule diseases patients were treated. 4 cases complicated hypertension and renal failure were failed to maintain their pregnancy. 6 cases whose glomerule disease were active but not complicated with hypertension and/or renal failure had their pregnancy successed. The results suggest that three factors should be put attention for pregnancy success in cases with concomitant chronic glomerule diseases. 1) Accurate evaluation of the criteria to allow pregnancy for patients with chronic glomerule diseases. 2) Immunosuppressive therapy should be maintained during pregnancy. 3) Regular follow up to detect and treat the complications promptly during pregnancy is never over emphysized. PMID- 9868037 TI - [The peripheral blood levels of TNF alpha and PLA2 activities in dialysis and non dialysis patients with uremia]. AB - The levels of tumor necrosis factor a (TNF alpha) and phospholipase A2 (PLA2) in peripheral blood were determined by means of the ELISA and improved micromethod in 31 patients with urmia and 12 normal healthy persons. The results showed: 1. In non-dialysis group and during a single dialysis session, a significant increase in the levels of TNF alpha was observed. 2. In non-dialysis and peritoneal dialysis patients, serum levels of PLA2 were significantly increased compared to normal subjects. The results suggest that acute complications induced by hemodialysis are related to increasing levels of TNF alpha, souble PLA2 in cleared by hemodialysis. PMID- 9868038 TI - [Multiple factors analysis of formation of corneal haze after photorefractive keratectomy]. AB - VISX 20/20 excimer laser was used to correct myopia from 1.00 to 16.00 diopters on 529 myopia eyes in 306 patients. All the patients were followed up for 3 to 12 months (8.1 +/- 3.1 months). At the 12th postoperative month, 0.5-3 grade corneal haze occurred in 62.1% of eyes in high myopia eyes, and only 8.8% in low and moderate myopia eyes (P < 0.01). The factors of formation of the corneal haze were the central corneal thickness, methods of removing epithelium and the number of laser pulses. The results suggest that the photorefractive keratectomy is regarded safe in correction of low and moderate myopia, but not in high myopia. PMID- 9868039 TI - [Study of arginase activity in alveolar macrophages from patients with lung cancer]. AB - Alveolar macrophages (AM) were obtained from 20 patients with primary lung cancer and 20 cases with nonmaligment pulmonary disease by BAL and were incubated in vitro in medium with and without BCG and/or IFN-alpha. Then the cell-free supernatants were harvested. The activity of arginase was assayed. It was found that: arginase was produced spontaneously by AM: The spontaneous production of arginase is lower in AMs from the tumor-bearing segments than either nontumor bearing segments or AMs from the patients with nonmalignant pulmonary diseases (P < 0, .05). The production of arginase was icreased on AMs from the tumor-bearing segments stimulated with BCG and/or IFN-alpha (P < 0.05 or 0.01). The results suggested that (1) There are some function at defects of tumor. (2) The mechanism of anti-tumor and anti-infection activity of BCG and IFN-alpha may possibly increase the production of arginase of AMs. PMID- 9868040 TI - [Internal fixation with U-shaped pin in the treatment of fracture-dislocation of the cervical spine]. AB - Twenty cases with fracture-dislocation of the cervical spine were treated by U shaped pin fixation. The U-shaped pin made of Steiman pin was used to hold sublaminar fixation of two segments above and below the vertebra of fracture. When atlas or axis was injured, a pair of small holes were drilled on the occipitale. The closed end of the U-Shaped pin was wired through the holes. Early postoperative ambulation was allowed without external fixation. 18 cases were followed up for 8-35 months. 5 cases of six patients with neurologic deficits recovered completely. The remaining one improved, no failure or loosening of the U-Shaped pin or wires was observed. Sound union of the bone grafts was obtained in all the patients. The results suggest that the operative procedure is simple, with sound fixation and satisfactory reduction and union of the fractures. PMID- 9868041 TI - [Broken head of a disc forceps left in the lumbar intervertebral space. A case report]. PMID- 9868042 TI - [CT features of hepatic cirrhosis in patients with chronic schistosomiasis]. PMID- 9868043 TI - [Effects of purerarin on blood superoxide dismutase activities and plasma malondialdehyde levels in patients with angina pectoris]. PMID- 9868044 TI - [cDNA library construction from compacted eight-cell mouse embryos and differential screening for specific-expressed cDNA clones]. AB - A large and representative cDNA library containing 6.9 x 10(5) independent clones was constructed from about 2 x 10(3) mouse embryos at the compacted 8-cell stage to identify and characterize gene products which play crucial roles in the first differentiation process of mouse embryogenesis. The cDNA library was differentially screened with labelled cDNA probes synthesized on poly (A)+ RNA isolated from the compacted 8-cell morula or late 2-cell mouse embryos. The results showed that difference in gene expression existed between these two developmental stages. Two clones which appeared only at the 8-cell stage were selected. This study provides a valuable tool for further detailed analysis of specific proteins associated with developmental events. PMID- 9868045 TI - [Optimum choice for the cultivation of CFU-GM in media buffered with HEPES]. AB - The effects of media buffered with HEPES of different concentrations and different incubation periods on the proliferation and differentiation of CFU-GM were studied under the standard condition for culture. The results exhibited that when the incubation period prolonged, the pH values of culture system raised and the colony formation of CFU-GM decreased; the culture system added with proper HEPES buffer solution could maintain and increase the colony formation of CFU-GM, especially the culture system in which the pH value was not adjusted before the experiment. We also found that media buffered with HEPES could change the ratio of different colony types. This showed that media buffered with HEPES could not only stimulate the proliferation of CFU-GM, but also affect its differentiation. The results suggest that media buffered with HEPES are better than those without HEPES for the culture of hematopoietic progenitor cells, and the optimum concentration is 10-20 mmol.L-1. PMID- 9868046 TI - [No point mutation of the 2.8 kb EcORI fragment of the nasopharyngeal carcinoma transforming gene TX in nasopharyngeal carcinoma]. AB - We have previously cloned a nasopharyngeal carcinoma transforming gene from human nasopharyngeal carcinoma cell line CNE-2 using mouse JB6 cell line as the recipient cells. This transforming gene, designated Tx, exhibits moderate transforming activity upon transfection into JB6 cells. Several lines of evidence indicate that the 2.8 kb EcoRI fragment which lies in the middle of the Tx gene is responsible for the transforming activity. The sequence of the 2.8 kb EcoRI fragment was determined, which showed striking homology with the human immunoglobulin light chain C region gene. Using the modified polymerase chain reaction-single strand conformation polymorphism analysis (PCR-SSCP) silver staining technique, we studied the possible point mutations of the nasopharyngeal carcinoma gene Tx in nasopharyngeal carcinoma. No mobility shift was detected in eleven paired cases of nasopharyngeal carcinoma biopsies suggesting that in most cases of nasopharyngeal carcinoma, the Tx gene is not activated via a mode of point mutation. Taken together with the previously obtained results we have found that the Tx gene is not activated by gene rearrangement or gene amplification. Therefore, we consider that the Tx gene may have been activated by its interaction with other regulatory factors. PMID- 9868047 TI - [Study on the cytotoxic and DNA damaging effects in oral mucosal fibroblasts by areca nut extract]. AB - An aqueous areca nut extract (ANE) was tested for its cytotoxic effects on cultured human embryo oral mucosal fibroblasts (HE-OMF) in vitro by using the trypan blue and thiazolyl blue (MTT) assay. The ANE decreases the cell survival rate in a dose-dependent manner (P < 0.05). So was the extract in inducing damage on the cellular DNA of HE-OMF in vitro examined by the nick translation assay. The increase in counts per minute (CPM) values was significant (P < 0.05) for comparing all four concentration groups tested, The results suggests that aqueous ANE is highly cytotoxic and capable to induce DNA damage on cultured HE-OMF. It may have potential carcinogenic effect on the oral mucosal membrane of whom habitually chewing the areca nut frequently for quite a long time. Futher study is required to illustrate the detail process and study the mechanism of these effects. PMID- 9868048 TI - [A study on detecting the gene deletion or loss of heterozygosity by polymerase chain reaction]. AB - The authors investigated the possibility of using PCR to dsetect the deletion of gene or loss of heterozygosity (LOH) of gene in biopsies. With some aids, such as selection of PCR cycle number, choice of internal control and screening of samples, the authors detected the deletion of GSH Clone 7 in nasophryngeal carcinoma (NPC) by PCR-Southern, and successfully found one case which partially deleted GSH Clone 7. This results was confirmed by image densitometry. Through this study, an available method to detect the deletion of gene by PCR is provided. PMID- 9868049 TI - [Establishment of mutant strain from human lung cancer cell (A549) with characteristics of resistance to 8-AG and HGPRT deficiency]. AB - A human lung cancer cell line (A549) mutant with hypoxanthine guanine phosphoribosyltransferase (HGPRT) deficiency and resistance to 8-AG was established by treatment of the A549 cells with 3 micrograms.ml-1 N-methyl-N' nitro-nitrosoguanidine (MNNG), and prescreened in 0.25% agarose DMEM semisolid medium containing 20 micrograms.ml-1 8-AG. The mutant cells are resistant to cytotoxic effect of 8-AG and sensitive to hypoxanthine-aminopterin thymidine (HAT) selective medium. The mutant cells can be used as a candidate parental cells to fuse with the somatic cells. PMID- 9868050 TI - [Effects of glutamate and ATP on interleukin 6 production by cultured neonatal rat astrocyte]. AB - The effects of glutamate (Glu) and ATP on interleukin-6 (IL-6) production from cultured neonatal rat astrocyte were studied. Glu (100-200 mumol.L-1) strongly increased, IL-6 production at 16 h in a dose-dependent manner and ATP (50-100 mumol.L-1) induced a inhibition effect. The results suggest that Glu stimulted IL 6 release may be one of mechanisms of astrocyte "neuroprotective" function in ischmia/reperfusion. PMID- 9868051 TI - [Effects of subretinal fluid containing proteins with different molecular weights on the proliferative vitreoretinopathy]. AB - The stimulating effects of subretinal fluid (SRF) containing different molecular weight proteins which were isolated by fast performance liquid chromatography (FPLC) and ultrafiltration on the proliferative vitreoretinopathy (PVR) were investigated. The results showed that (1) the PVR was induced by 10-30 kD proteins injecting vitreously, and the forming duration of PVR was about two weeks; (2) the location of inducing PVR was mainly on medullary ray area in the rabbit eye; (3) In the SRF there were growth factors which were mainly those proteins with molecular weight between 10 kD and 30 kD. PMID- 9868052 TI - [The study of acute clinical electrophysiological effects of propafenone on paroxysmal supraventricular tachycardia using transesophageal atrial pacing technique]. AB - Trans-esophagus atrial pacing (TEAP) was employed to evaluate the acute electrophysiological effects of propafenone administrated intravenously in 50 patients with SVT. Forty three of the subjects are patients with atrial ventricular reciprocating tachycardia (AVRT). Seven of them were involved in atrioventricular nodal reentry tachycardia (AVNRT). The results indicate that propafenone exerts an obvious inhibitory effect on both the dual atrioventricular node pathways and the accessory pathways, with the latter one being markedly affected. In addition, propafenone can deter the antegrade and retrograde conduction and prolong the refractory period of the accessory pathways. Still more, it results in preceding of the Wenchebach and 2 to 1 block point of the atrioventricular node conduction. All these promise the potential terminating effect on tachycardia. Propafenone has little effects on sinus node. Prolongation of sinus node recovery time (SNRT) to 3600 ms following drug administration was observed in only one patient who has a history of sinus bradycardia. PMID- 9868053 TI - [The sonarographic diagnosis of prostatic and periprostatic cysts on high resolution transrectal ultrasound imaging]. AB - The symptoms of prostatic and periprostatic cysts are nonspecific and digital rectal examination has no definite findings detected. The conventional radiological plain film cannot define the relationship of a cyst to the surrounding structures. The use of MRI as a means in detection is too expensive. To solve these problems, we detected 118 cysts in 73 patients by transrectal ultrasound (TRUS) and discussed the TRUS imaging findings of various prostate and periprostate cysts with illustrations from our clinical practice. We suggest that TRUS can be used routinely, because its distinct advantage of high resolution for minute nodular or cystic lesion detection with high quality image demonstrates the relationship of surrounding structures, and provides an outstanding novel diagnostic modality. PMID- 9868054 TI - [A preliminary evaluation on transcranial Dopplor ultrasonic scanning for differentiation of symptoms and signs in patients with ganyang shangkangzheng]. AB - Intracranial main artery of Ganyang Shangkangzheng and its related syndrome were determined by Transcranial Doppler (TCD) Ultrasonography. The results showed that the blood flow speed in the mid-cerebral artery of 32 cases with Ganyang Shangkangzheng was obviously higher than that of the healthy group, Ganhuo Shangyan group and Gan-shenyinxu group, similar change was found in three kinds of parameters in the blood flow speed. Therefore, the results suggest that TCD in determinating the blood flow speed of middle cerebral artery can be thought as one of the diagnostic assays in differentiation of symptom and sign of Ganyang Shangkangzheng. PMID- 9868055 TI - [The application of intracoronary stenting to type B and C lesions of coronary arteries]. AB - Intracoronary stenting was introduced to 4 lesions due to coronary arterial dissections and to 2 lesions owing to elastic recoil of coronary arteries and stent placement was directly performed in 6 complex coronary lesions after 12 target lesions from 7 patients with Types B and C lesions of coronary arteries had been pre-dilated with balloon. Coronary angiography showed that dissections and recoils had disappeared. Target lesions were well dilated and blood flow was well improved in all cases, one patient was complicated with hemorrhage which was cured quickly. During the follow-up period of 1 to 24 months, another case had recurrence of angina after 2 months of stenting which could be controlled with anti-angina agents. These indicate that intracoronary stenting had good therapeutic effect and is considered as a safe, fast and effective method in dealing with acute complications of coronary artery after routine PTCA. PMID- 9868056 TI - [A clinicopathological immunohistochemical and cytomorphometric study on midline T cell lymphoma]. AB - One hundred and twenty-two cases of midline malignant reticulosis (MMR) were studied. A series of antibodies including anti-LCA, UCHL-1, L26, CD45R, and anti lysozyme were used on paraffin sections by ABC and PAP methods. The results were as follows: 112 cases exhibiting T-cell origin, 4 cases showing B-cell origin, and 6 cases being of uncertain lineage. This result is in accordance with the point of view that most of MMR are T cell lymphoma. Two histological types were classified: sarcomatoid type and granulomatoid type. By using image analyzer, the sarcomatoid type was subdivided into small, medium and large cell types. Sixty two cases with follow-up data were collected for clinicopathologic analysis. One year and five-year survival rates in cases with different histologic types were compared and statistically analysed. The results showed that the prognosis was closely related to the histological type. PMID- 9868057 TI - [Evaluation of optimal measuring site and index by QDR4500A for postmenopausal bone loss]. AB - As the function of most bone mineral measuring machine is limited, either by single photon absorptiometry (SPA), dual photon absorptiometry (DPA), speed of sound (SOS), or dual energy x-ray absorptiometry (DEXA), it is hard to evaluate the best place and index of postmenopausal bone loss at many positions by one machine at the same time. So we used QDR4500A, one of the best advanced DEXA which has a good repeatability, to search the best place and index for postmenopausal bone loss. The results showed that (1) bone mineral density (BMD) was better than bone mineral content (BMC) and Area to reflect this kind of bone loss; (2) lumber BMD was a better place than the forearm BMD, total spine BMD and other places; and (3) midlateral L3, 4 BMD was a best index for postmenopausal bone loss. PMID- 9868058 TI - [A study on the infiltration of macrophages and lymphocytes in interrenal tissues in patients with acute renal tubulointerstitial disease]. AB - Renal tissues of 14 patients with acute renal tubulointerstitial disease were studied by using immunohistochemical method. Cells infiltrated in interrenal tissue were observed. The results show that the causes which result in acute renal tubulointerstitial disease are different; the renal tubular and interstitial damages are reversible or irreversible; the major inflammatory cells infiltrated in interrenal tissues are CD68 and CD45ro positive cells. The results suggest that the macrophages and T lymphocytes infiltrated in interrenal tissues play an important role in the progress of acute renal tubulointerstital disease. PMID- 9868059 TI - [Effects of smoking on oxygen free radicals and fibrinolytic system]. AB - The effects of smoking on SOD, MDA, t-PA, PAI-1 were studied in 20 healthy men (10 smokers and 10 nonsmokers). RESULTS: SOD decreased and MDA increased significantly in smokers. After acute smoking, SOD decreased and MDA increased significantly in all the subjects, t-PA and PAI-1 activities were increased immediately after acute smoking. There was a significant positive relationship between increased t-PA and SOD, whereas a significant negative relationship exists between increased t-PA and MDA. CONCLUSION: Smoking increases oxygen free radicals. The release of t-PA is decreased in smokers, that might be resulted from the increases of oxygen free radicals. PMID- 9868060 TI - [Electromyogram (EMG) and muscle biopsy in patients with chronic spinal muscular atrophy]. AB - The clinicopathological changes were investigated by electromyogram (EMG) and muscle biopsy in 30 cases of chronic spinal muscular atrophy. The results showed that the denervating changes were demonstrated in EMG in all patients but two. The light microscope discovered manifestation of neurogenic atrophy, and the electron microscope revealed a reduction of myofibrils, thickening and wavy appearance of Z-lines and dilation of mitochondria and endoplasmic reticula. PMID- 9868061 TI - [The value of electronystagmography in differential diagnosis of vertigo]. AB - Findings of electronystagmography (ENG) in 645 patients with vertigo were analysed. Two hundred and twelve cases showed vestibular peripheral vertigo, 394 cases vestibular central vestigo, and 39 cases other types of vertigo. The ENGs in patients with vestibular peripheral vertigo were charazterized mainly by the vestibular abnormal caloric responses, spontaneous and benign positional nystagmuses were also obsedrved at the same time. Different degrees and various combinations of the abnormal responses shown in optokinetic central test were the most characteristic of ENG in patients with vestibular central vertigo. The results suggest that ENG is particularly useful for differentiating peripheral and central vertigo. PMID- 9868063 TI - [Pre- and post-operative changes in serum levels of glycocholic acid and 3,5,3' triiodothyronine and their clinical significance in patients with cholelithiasis]. AB - Pre- and post-operative changes of serum glycine-conjugated cholic acid (CG) and 3,5,3'-triiodothyronine (T3) levels were observed in twenty-two patients with cholelithiasis. The increase of the levels of serum CG varied with the patients; the highest levels were seen in patients with cirrhosis. After operation, the levels of serum CG and T3 were decreased significantly in patients without cirrhosis. There was positive correlation between serum CG and T3 (r = 0.4667, P < 0.01). It is indicated that the lowering of serum CG after operation may be related to the lowering of serum T3; the significant increase of serum CG level is useful to the diagnosis of liver cirrhosis, which is more sensitive than Type B ultrasonography or routine examination of liver function. PMID- 9868062 TI - [Changes of early renal function before and after using mannitol in patients with cerebral apoplexy]. AB - The alteration of renal function after initiation of mannitol infusion were monitored in 20 patients with cerebrovascular accident to evaluate whether mannitol has nephrotoxicity effects. Serum creatinine and urea nitrogen were increased after initiation of mannitol infusion, but hadn't reached stagistical significance. Urinary and serum alpha 1-microglobulin, beta 2-microglobulin and urinary NAG, gamma-GT were significantly increased after 5-10 days successive infusion of mannitol (P < 0.05). The results suggest that the acute nephrotoxicity effect of mannitol should not be ignored. PMID- 9868064 TI - [Clinical analysis of 120 patients with fibromyalgia]. AB - The clinical characteristics of 120 patients with fibromyalgia is reported. Most of the patients were 21-50 years old. More than 56% patients had three chief symptoms: generalized myalgia, abnormal sensations and easily fatigue. 49.2% of the patients complained multiple arthralgia; 29% with sleep disorder and 23% with local myospasm of lower limbs. The trigger point was regarded as an important physical sign. The etiology and differential diagnosis for fibromyalgia were discussed. PMID- 9868065 TI - [Clinical and laboratory studies with typhoid fever 178 patients]. AB - One hundred and seventy-eight cases of typhoid patients were studied on clinical and bacteriological aspects. The main clinical findings were as follows: (1) Most of the cases had sustained fever (66.3%). (2) Gastroenterial symptoms developed as the disesase progressed. (3) Rose spots were found in 32.6% of them. (4) Liver and spleen were enlarged in 69.5% of the cases. (5) Blood eosinophil disappeared in most of the patients and leukopenia was noted in 94.3%. (6) There were toxic hepatitis (47.1%), toxic myocarditis (22.4%) and intestinal hemorrhage (19.7%) as complications. In the drug sensitivity test, the number of ampicillin-resistant and chloramphenicol-resistant strains of salmonella typhi was increased more than that seen 5 years ago (P < 0.05), however 100% of the strains were sensitive to amikacin, tobramycin, norflexacin, oflexacin and the third generation of the cephalosporin. For the time being, norflexacin and oflexacin were good and suitable drugs for the treatment of typhoid fever. PMID- 9868066 TI - [Electroencephalogram changes of meningioma]. AB - Electroencephalogram (EEG) was used in investigating 42 cases of meningiomas proved by operations and pathological examinations. The abnormality rate of EEG was 95.24%. Compared with the clinical findings, the accuracy rate of EEG lesion location was 64.29%; the accuracy rate of EEG lesion-side location was 26.19%; the total accuracy rate of lesion-location and lesion-side location was 90.48%. The EEG characteristics and the value of location diagnosis of meningioma were discussed. This indicates that EEG is still an important supplementary diagnostic method of intracranial meningioma. PMID- 9868067 TI - [Harmful, non-effective and benign dislodgements of electrodes in permanent right ventricular pacing]. PMID- 9868068 TI - [Significance of antinuclear antibodies for the diagnosis and activity of systemic lupus erythematosus]. PMID- 9868069 TI - [Establishment of hybridoma technique and study on fusion conditions of hybridoma cells]. PMID- 9868070 TI - [Effect of nitric oxide and tumor necrosis factor on pericardial effusion]. PMID- 9868071 TI - [One type of new glycophorin variant found in Hunan]. PMID- 9868072 TI - [Relation between oxygen saturation of right atrial blood and cardiac index]. PMID- 9868073 TI - [Diagnostic error of congenital diaphragmatic hernia in 11 children]. PMID- 9868074 TI - [A case of subacute bacterial endocarditis caused by anaerobic Peptostreptococcus]. PMID- 9868075 TI - Effects of losartan and PD123319 on antigensin II-induced proto-oncogene expression and protein synthesis in cultured neonatal rat cardiac myocytes. AB - To study the effects of specific angiotension II (Ang II) receptor, type-1 (AT1) antagonist (Losartan) and type-2 (AT2) antagonist (PD123319), on Ang II-induced proto-oncogene expression and synthesis of RNA and protein in neonatal rat cardiac myocytes, we used respectively [3H]-uridine and [3H]-leucine incorporation to measure the rate of RNA and protein synthesis, and analyzed the c-myc mRNA level by Northern blot. We found that an acceleration in the rate of RNA and protein synthesis was observed when exposed to 2.5 x 10(-6) mol.L-1 [Sar1] Ang II for 24 h (P < 0.01). Losartan inhibited the action in a dose dependent manner. The level of c-myc mRNA was up-regulated to 340% of control by 2.5 x 10(-6) mol.L-1 Ang II, and Losartan (10(-5) mol.L-1) suppressed the increase of c-myc mRNA stimulated by Ang II. PD123319 showed similar inhibition on Ang II-induced RNA and protein synthesis, but did not inhibit c-myc expression. Thus, Ang II-stimulated expression of c-myc is mainly mediated by AT1 receptors, and contributes to cardiac myocyte hypertrophy while AT2 receptors are involved in mediation of cellular growth without altering of c-myc expression. PMID- 9868076 TI - [Artificial biological skin for preventing adhesion of repaired tendon in white rats]. AB - To study the effect of tendon healing and adhesion after tendon repaired using the artificial biological skin (Biofill) produced in Brazil, we divided randomly 46 rats [correction of mice] into groups, compared the pelvic limbs with themselves, observed by eyes and light microscopy, and made biomechanical and histological detection in 1st, 2nd, 3rd and 4th weeks postsurgically. The results showed that the speed and strength of tendon healing had light affect in three weeks after operation, and had no difference between experimental groups and control groups after 4 weeks. It is indicated that artificial skin "Biofill" can effectively prevent tendon adhesion after tendon repaired and have not obviously influenced in the intrinsic self-healing process of tendon. PMID- 9868077 TI - [Preliminary evaluation on bone mass of tibia by ultrasonic measurement]. AB - The speed of ultrasound (SOS) in right middle tibia and bone mineral density (BMD) of left radius at the distal one-third were measured by ultrasonic measurement and single photoabsorptionmetry in 260 healthy individuals simultaneously. SOS of tibia was positively correlated with BMD of radius (r = 0.64). The peak bone mass of both measurements was at 33 to 36 years of age. Height was positively correlated with the SOS of tibia, while weight was positively correlated with SOS of female tibia, period of menopause negatively correlated with female's SOS of tibia. The concordance of SOS of tibia and BMD of radius in diagnosis of osteoporosis was 36.8%. The results suggest that ultrasonic measurement of tibia is an effective method of bone mass measurement. PMID- 9868078 TI - [Effects of sevoflurane on contractility of isolated thoracic aorta rings in rabbits]. AB - The objective of this paper was to observe the effects of sevoflurane on isolated rabbit aortic rings and the mechanisms of the effects. Twenty-four vascular rings collected from different New Zealand rabbits were randomly divided into 4 groups: Group 1 endothelium-inact rings; Group 2 endothelium-denuded rings; Group 3 endothelium-intact rings plus indomethacin; Group 4 endothelium-denuded rings plus indomethacin. When stable active tension produced by phenylephrine (1 x 10( 6) mol.L-1) was obtained, sevoflurane was added into the tissue bath. The anesthetic gas was increased in 0.5% increments every 10 min until the 4% of sevflurane after which 5% was administered. In rings of Group 3 and Group 4, indomethacin (2.8 x 10(-5) mol.L-1) was added to the bath 10 min before the addition of sevoflurane. The results showed that sevoflurane relaxed all groups of rings dose-dependently and this action was non-endothelium dependent and indomethacin had no effect on it. It suggests that sevoflurane relaxes peripheral vessels perhaps by affecting vascular muscle directly. PMID- 9868079 TI - [Effect of monoclonal antibody against human tissue factor pathway inhibitor on plasma coagulation time]. AB - The purpose of this study was to evaluate the influences of the monoclonal antibody against human tissue factor pathway inhibitor (TFPI), McAb4F8, on prothrombin time (PT) and activated partial thromboplatin time (APTT). The results showed that PT and APTT were prolonged by exogenous TFPI. Dilute thromboplastin coagulation time was shortened with a dose-dependent manner by McAb4f8. Low molecular weight heparin (LMWH) prolonged significantly PT and APTT, and this effect ws weakened by an antibody against human AT III. McAb4F8 was also proved to shorten dilute thromboplastin coagulation time of factor IX deficient plasma. These results indicated that the tissue factor pathway plays an important role not only in physiological coagulation but also in hemorrhage of hemophilia. PMID- 9868080 TI - [Delayed brain injury induced by Bordetella pertussis in rats]. AB - To induced new model of delayed infection brain injury, we divided randomly 40 adult male Sprague-Dawley rats into four groups: bordetella pertussis (BP) groups, and normal sailine (NS) groups in 4 h (n = 10) and 24 h (n = 10), respectively, injected BP at 0.2 ml.kg-1 which contained the bacilli in 10.8 x 10(9).ml-1 into the rat left internal carotid artery of BP groups, and injected at the equal volume as control in the same way into NS groups. The water content (WC), Evans blue content (EB), cations of sodium, potassium, calcium in the brain tissues were measured and the morphologic changes in the hippocampal CA1 were detected by electron microscopy. These results were shown as follows: the left hemisphere of BP group in 4 h was characterized by the brain edema which predominated in swollen, necrotic astroglial cells, endothelial cells, and insignificant increase of contents of sodium, calcium and decrease of potassium; BP group in 24 h was characterized by dark cells, the brain edema also existed in astroglial cells. These results indicate that a delayed infection brain injury model is performed successfully by injection with BP into the left internal carotid artery of rats. The model is characterized by the features of delayed neuronal death and mixed type brain edema. PMID- 9868081 TI - [Effects of aprotinin on heparinized whole blood activated clotting time and whole blood prothrombin time]. AB - When aprotinin is used during cardiopulmonary bypass, there is a prolongation of the activated clotting time (ACT), which is used to monitor heparinization. The aim of this study was to observe the effects of aprotinin and heparin on whole blood ACT and whole blood prothrombin time (BPT). The results showed that when kaolin was used as the contact activator, the intrinsic clotting system was also inhibited by aprotinin, the observed ACTs with various dose aprotinin and concomitant heparin were significantly prolonged (Q = 0.757, P < 0.01). There was a dose-dependent prolongation of BPT by heparin (r = 0.985, P < 0.01). However, the heparin-mediated prolongation of BPT was not enhanced by aprotinin. The authors conclude that aprotinin prolongs heparinized whole blood activated clotting time but was not whole blood prothrombin time. PMID- 9868082 TI - [Mitomycin C concentrations in rabbit ocular tissues after topical administration during glaucoma filtration surgery]. AB - Using high-performance liquid chromatography, we measured Mitomycin C (MMC) concentrations in conjunctiva, sclera and aqueous of 22 rabbit eyes after single topical administration of 0.2 mg.ml-1 MMC during glaucoma filtration surgery. The peaks of MMC concentrations in conjunctiva, sclera, aqueous were 2.01 micrograms.g-1, 2.95 micrograms.g-1 and 0.16 microgram.ml-1 with half life of 0.63, 0.35 and 0.84 hours respectively. Irrigating the ocular surface with 20 ml of normal saline after MMC application reduced the peak drug concentration to 1/8 in conjunctiva, to 1/5 in sclera and to 1/3 in aqueous. The results showed that the MMC concentrations in conjunctiva and sclera were well above the ID50 of rabbit subconjunctival fibroblast (0.1 microgram.ml-1), and the concentration in aqueous was well below the level known to cause endothelium toxicity (approximately 0.2 mg.ml-1) and retinal toxicity (> 1.3 micrograms.ml-1). Therefore, 0.2 mg.ml-1 MMC can inhibit subconjunctival fibroblast effectively and has no side effect on visual function. PMID- 9868083 TI - [Comparative study of expression of p21 and c-myc protein in hepatocellular carcinoma with pericarcinomatous liver tissue]. AB - Expression of p21 and c-myc protein in hepatocellular carcinomas and their surrounding liver tissue was detected on serial sections by immunohistochemical method. The results showed that the positive rates of p21 expression were 53.3% (16/30) and 96.7% (29/30) in hepatocellular carcinomas and pericarcinomatous liver tissue, and 40% (12/30) and 86.7% (26/30) and 86.7% (26/30) for c-myc protein expression respectively. Their incidences in pericarcinomatous liver tissue were higher than that in cancer tissue (P < 0.01). The patterns of p21 and c-myc protein in cells were cytoplasm, membrane and/or nuclear types. Their expression was more intensive in pericarcinomatous hepatocytes, especially in liver cirrhosis nodes. The results indicate that abnormal activiation and expression of oncogene ras and c-myc may be related to hepatocellular carcinogenesis. PMID- 9868084 TI - [A histopathological study on Hunan aqueous drainage implantation in rabbit's eye]. AB - Thirty eyes with 15 healthy white New Zealand rabbits subgrouped into two groups: right eye as an experimental group which was implanted Hunan aqueous drainage (HAD) and left eye served as self-control group in which a silicone band 7 mm in width and 7 to 10 mm in length was placed equatorially around the globe. Within 1 to 12 weeks postoperatively, we examined the anterior segment of the globe by the slit lamp microscopy (SLM) and made histopathological observation under light microscope after sacrifice and enucleation of eyeballs in five batches. SLM examination showed that 8 out of 15 eyes in experimental group showed mild or moderate ocular reaction which subsided within 2 to 14 days postoperatively. Histopathological observation revealed no obvious pathological lesions in the uveal tract tissues in the most globes. No foreign body macrophages were found. It indicates that HAD implant is nontoxic and nonallergenic, so it is adaptable for rabbit ocular tissue. The HAD implant can be used in human eyes, too. PMID- 9868085 TI - [The spontaneous otitis media in rickety rats]. AB - This paper is aimed to study the incidence and hearing impairment of spontaneous otitis media in rickety rats during a longterm vitamin D deficient breeding. Thirty Sprague-Dawley rats ranged from 20 to 25-day-old were used, and vitamin D deficient rickets models were established. Another thirty rats were used as the normal control. It was found that both normal and rickety rats had the possibilities of suffering from otitis media (mainly exudative) during the two month-breeding. The incidence of otitis media in normal rats was 5% (3 from 60 ears), while that in the rickety ones was 16.67% (10 from 60 ears). It is believed that a rickety sufferer may be in higher danger of having otitis media than the normal one. PMID- 9868086 TI - [Lipoprotein(A) distribution among adults and its relation to the incidence of coronary artery disease in Changsha City]. AB - The plasma lipoprotein (a) [Lp(a)] levels were determined among 593 adults in Changsha city, Hunan province. The result showed that the distribution of Lp(a) levels in all the individuals was a skewness which can be normalized by data transformation into logarithm. Plasma Lp (a) concentration > 300 mg.L-1 was 34.8% in 209 subjects with coronary artery disease (CAD) and 17.3% in 305 health controls (P < 0.01). The Lp(a) levels were significantly higher in subjects with CAD than in health controls. Multiple regression stepwise analysis found that Lp(a) levels were positively correlated with CAD. This study indicates that plasma levels of Lp(a) are strongly related to CAD and may be an important independent risk factor for CAD. PMID- 9868087 TI - [The changes of PRA, ATII, ald, ET and ANP in patients with left ventricular diastolic heart failure and intervention with enalapril]. AB - Fifty patients with left ventricular diastolic heart failure (LVDHF), and 35 patients with left ventricular systolic heart failure (LVSHF) diagnosed by clinical manifestation and radionuclide ventriculography were studied and 20 normal persons served as controls. Plasma renin activity (PRA), angiotensin I (AT I), aldosterone (ALD), endothelin (ET) and atrial natriuretic peptide (ANP) concentrations were measured by radioimmunoassay. Fifty patients with LVDHF were divided into treatment group and control group in a randomized, double blind, control method. Enalpril, CoQ10 and VitE were given in treatment group while only CoQ10 and VitE were given in control group. The therapeutic efficacy was evaluated after 8 weeks of treatment. The results showed that plasma concentration of PRA, AT I, ALD, ET and ANP were increased in LVDHF, but lower than those in LVSHF. After treatment with enalapril plasma PRA was increased while AT I, ALD and ET level were decreased significantly but ANP level had no change in treatment group. PMID- 9868088 TI - [Correlation between shape and direction of small articular surface in lower lumbar vertebrae and degeneration of intervertebral disc]. AB - To assess the possible correlation between the shape and the direction of the small articular surface in the lower lumbar vertebrae and the degeneration of the intervertebral disc, we investigated with computed tomography (CT) and evaluated with statistics the small articular surface and the transverse interface-joint angle (TIFA) of the L4-5 and the L5-S1 in 152 cases who had normal or degenerative discs verified through CT, MRI or operation. The small articular surface was found arc in 69.1% of the L4-5 and in 23.0% of the L5-S1. The TIFA of the L4-5 was less than that of the L5-S1. There was no correlation between the ratio of degeneration of the intervertebral disc at the L4-5 and the TIFA of the L4-5 and the L5-S1, but the ratio of degeneration of the intervertebral disc at the L5-S1 had postive correlation with the TIFA of the L4-5, negative correlation with the TIFA of the L5-S1, and particular correlation with the TIFA of the L5-S1 and L4-5. These results suggest that the shape and direction of the lower lumbar facet joint are related to the lumbar degeneration of intervertebral disc and the causes of degeneration at the L4-5 disc differ from those at the L5-S1 disc in biomechanics. PMID- 9868089 TI - [The mediopatellar plica syndrome of knee in 61 cases]. AB - From 1989 to 1996, 61 patients, including 66 knees, aged 8 to 62, were diagnosed as mediopatellar plica syndrome of knee after arthroscopic examination. Twenty one cases were misdiagnosed as medial meniscal lesions before arthroscopic examination. Fifty-nine patients (64 knees) underwent arthroscopic resection of the plicae and 2 patients underwent opened resection. Fifty-nine cases had an average follow-up of seven months. Fifty cases (54 knees) of them showed excellent and good results. PMID- 9868090 TI - [Change in nitric oxide contents of knee joint fluid in patients with degenerative osteoarthritis]. AB - The degenerative osteoarthritis (DOA) is a common disease of aged people. But its pathogenesis and mechanism are not very clear. We compared the Nitric Oxide content in knee fluid of DOA with the normal people as matched age. The results indicated that the Nitric oxide has a high concentration in DOA. This suggested that the Nitric Oxide may play an important role in the mechanism of this disease. PMID- 9868091 TI - [A study on the relationship between plasma level of ET-1 and cGRP and blood pressure in hemodialysis patients]. AB - Plasma endothelin-1 (ET-1) and calcitonin gene related peptides (cGRP) were studied by immunoradiological method in 60 hemodialysis (HD) patients. The results showed that: (1) Plasma level of ET-1 and cGRP were increased in the HD patients. (2) Plasma level of ET-1 and cGRP varied in different times of HD. (3) A significantly positive relation between plasma ET-1 and blood pressure (BP) existed in the HD patients, while a significantly negative relation between plasma cGRP and BP existed in the HD patients. The results suggest that ET-1 and cGRP are major factors which lead to BP changes in HD patients. PMID- 9868092 TI - [Detection of p53 gene mutations in human gestational trophoblastic neoplasia by polymerase chain reaction-single strand conformational polymorphism techniques]. AB - Using polymerase chain reaction single strand conformational polymorphism (PCR SSCP) and EB staining technique, paraffin-embeded sections of 20 hydatidiform mole and 4 choriocarcinoma were detected in the exons 5 and 8 of p53 gene. The results showed that mutations of p53 gene were 0/9 in the normal chorionic villi, 6/20 (30%) in hydatidiform mole and 3/4 in choriocarcinoma. This study suggests that mutations of p53 gene may be an important event in human gestational trophoblastic neoplastigenesis and its progression. PMID- 9868093 TI - [Radiofrequency catheter ablation of idiopathic ventricular tachycardia]. AB - Thirteen consecutive patients with idiopathic ventricular tachycardia underwent radiofrequency catheter ablation. This group included 9 idopathic left ventricular tachycardia (ILVT) and 4 idiopathic right ventricular tachycardia (IRVT). Five ILVT patients with left axis deviation and one with right axis deviation were ablated successfully. By pace mapping, two IRVT patients with ventricular tachycardia originating from right ventricular out-flow tract were ablated. No complications occured. By means of follow-up of 3-22 months one case showed recurrence with successful reablation. It indicates that radiofrequency catheter ablation therapy is an effective and safe procedure in patients with idiopathic ventricular tachycardia. PMID- 9868094 TI - [Expression level of MDR1, and MRP in patients with malignant blood disease and its clinical significance]. AB - RT-PCR method was used to detect the expression level of multi-drug resistance gene (MDR1) and multi-drug resistance associated protein (MRP) in patients with malignant blood disease, including 19 cases of acute leukemia and 6 cases of multiple myeloma. The results showed: expression level of MDR1 in patients with known clinical drug resistance elevated obviously and was significantly associated with clinical drug resistance (r = 0.612, P < 0.01), which indicated that expression of MDR1 was the main mechanism of clinical drug resistance. There is no obvious relationship between expression level of MDR1 and MRP (r = 0.035, P > 0.05). PMID- 9868095 TI - [Therapy for intracranial lesions by gamma knife]. AB - The authors summarized 281 patients with intracranial lesions treated by gamma knife (GK) during Sept. 1995-Sept. 1996. The patients included 170 males and 111 females with age ranging from 5 to 85 years (average of 49.9 years). Seventy seven patients received a follow-up 3.3-12.5 months. The average of Karnofsky Performance Status was 70.8 in pre-operation and 83.9 in post-operation; the average of tumor volume was 17.1 cm3 in pre-operation and 13.6 cm3 in post operation; the average of lesion diameter was 2.8 cm and 2.4 cm in pre- and post operation. respectively. GK radiosurgery has achieved the goal to improve the quality of life and to prolong the life of those patients with malignant intracranial tumors. PMID- 9868096 TI - [The analysis of Q-Tc interval changes in treadmill exercise-induced myocardial ischemia]. AB - The Q-Tc interval before and after the treadmill exercise test in 48 normal subjects and 59 patients with coronary heart disease were compared. The Q-Tc interval was shorter in normal group at immediate moment, 2, 4 and 6 minutes after the exercise than before. In the coronary heart disease group, the Q-Tc interval at the same time points as above was longer after exercise compared to pre-exercise. There was no significant difference of exercise-induced Q-Tc changes in silent and symptomatic myocardial ischemia. PMID- 9868097 TI - [Development of three dimensional bone-connecting plate and its clinical application]. AB - Biomechanical assessment showed that the three dimensional plate developed by the authors which had stereo-rigidity in the fixation of long bone fractures was superior to compressive plate and flanged plate in terms of anti-compressive and anti-torsional strength. The anti-bending strength was superior to the flanged plate but inferior to compressive plate. The comprehensive nature of the three dimensional plate remained superior. Clinical application in 112 cases with 85 of them being followed-up for 8-44 months showed that bone union occurred in 45-98 days. The average was 82 days. In 65 cases of them the plates had been removed. No delayed-union or non-union occurred. Preliminary experience showed that the three dimensional plate was a new design for the treatment of long bone fractures. It can be applied to various types of long bone fractures especially the severe comminuted fractures owing to its requirement of proper fixation. PMID- 9868098 TI - [The value of detecting CVB-IgM antibodies in diagnosing viral myocarditis of children]. AB - By using the indirect immunofluorescence assay and the tissue cells infected with coxackie virus Group B as antigen, the authors detected the antibodies type IgM to coxsackie virus Group B (CVB-IgM) of the sera of 105 children with viral myocarditis (VMC), 59 children with other diseases (COD) and 67 healthy children (HC). The results showed that in VMC, the genometric mean titre (GMT) of CVB-IgM in female (1:16.58) was higher than that in male (1:9.28, P < 0.05); not only in female, but also in male the GMT of VMC was higher than that of HC; also the GMT in female of VMC was higher than the GMT in female of COD, but no considerable difference existed between male of VMC and COD. Without sex distinction the GMT of HC was 0.289, its standard variance was 0.9335, the upper limit titre of normal range was 1:8.6. The authors took 1:10 as the positive criterion, the sensitivity of CVB-IgM in diagnosing VMC was 79.05%, the specialities of differentiating VMC from COD and HC were 42.3% and 91.04%, the consistancy rates of diagnosing VMC from COD and HC were 65.85% and 83.72% respectively. PMID- 9868099 TI - [Simultaneous determination of eight kinds of conjunct bile acids in human bile by R-HPLC]. AB - A method for the simultaneous determination of eight kinds of conjunct bile acids in human bile was developed by HPLC. They were separated on a YWG-C18 (3 microns) column at 30 degrees C, with methanol/water (65/35, V/V, pH3.0) as mobile phase, and detection wavelength at UV 210 nm. The linear ranges were 50-1,000 microns.ml 1, the recoveries were 91.2%-108.6%. The biles of 30 cases with cholelithiasis cholecystolithiasis and 20 cases without gallstone were detected by HPLC. The results showed that the constitution of bile acids was different between patients with cholelithiasis cholecystolithiasis and patients without gallstone. PMID- 9868100 TI - [Determination of the content for micronomicin sulfate by ultraviolet spectrophotometric derivatization]. AB - An ultraviolet spectrophotometric method for determination of micromonicin (MCR) was developed by the reaction of MCR with o-pathaldehyde (OPA) and obtained a derivative absorbing ultraviolet in special wave length. The comparison was made between this method and high performance liquid chromatography (HPLC). The results indicate that the method is simple, rapid, precise and reproducible. PMID- 9868101 TI - [Ultrasonography in the diagnosis of heart carcinoma metastasis from liver cancer. Report of 2 cases]. PMID- 9868102 TI - [Measurement of area values of cardiac impedance differential loop in different age groups]. PMID- 9868103 TI - [Short-term observation on the therapeutic effects of percutaneous balloon mitral valvuloplasty]. PMID- 9868104 TI - [Diagnosis and treatment of retroperitoneal chemodectoma. Report of 7 cases]. PMID- 9868105 TI - [Clinical significance of combined detection of 7 kinds of antinuclear antibodies with immunoblotting techniques]. PMID- 9868106 TI - [Successful replantation of whole scalp avulsion with micro-vascular anastomosis. A case report]. PMID- 9868107 TI - [Changes in serum cortisone levels in patients with non-insulin-dependent diabetes mellitus]. PMID- 9868108 TI - [Endocardial endothelium modulates lysophosphatidylcholine induced positive inotropism]. AB - In isolated guinea pig papillary muscles, the endocardial endothelium (EE) was selectively damaged by exposing the muscles to a flow of dry air for 30s for removal of function EE. The results showed that LPC induced positive inotropic effect on papillary muscles was observed in a concentration-and time-dependent manner [PT after 15 mins' exposure to 5, 10 and 20 mumol.L-1 lysophosphatidylcholine (LPC) was increased by 104 +/- 7%, 110 +/- 6% and 124 +/- 10%, respectively]. The positive inotropic effect of 10 mmol.L-1 LPC on papillary muscles denuded of EE was significantly enhanced (PT: 163 +/- 23% vs 130 +/- 17% after exposing to LPC for 30 min). Scanning and transmission electronic microscopic studies exhibited signs of EE damage of papillary muscles incubated with LPC 10 mumol.L-1 for 30 minutes, such as disruption of gap junction between overlapping EE cells, retraction of the cell borders, fenestration on the EE, and denudation of the subjacent basal lamina. The present study suggests that the EE may affect the LPC-induced contractibility of papillary muscle via its barrier function signs, in addition, the present data provides further evidence that LPC may be a trigger for some kind of biological active factor release form EE. PMID- 9868109 TI - [Activation of L-Arg: no pathway in canine brain by the damage from complete cerebral ischemia-reperfusion]. AB - To ascertain whether complete cerebral ischemia-reperfusion activate L-Arg: NO pathway in canine brain, we anestherized nine adult dogs with ketamine and fentayle and randomly divided into two groups. Four dogs were nonischemic control group. Five dogs were complete cerebral ischemia-reperfusion group, they underwent a 18-minute cardiac arrest, and were resusciatation by standard CPR, supported by intensive care for 8 hours. At the end of each experiment, the parietal cortex was assayed for content of Nitrite and NADPH-positive neurons. Compared with the control group, the contents of Nitrite and NADPH-positive neurons of coxtex in complete cerebral ischemia-reperfusion group increased significantly (P < 0.01). The results suggest that complete cerebral ischemia reperfusion activate the L-Arg: NO pathway in canine brain, and NO may play an important role in cerebral ischemia-reperfusion injury. PMID- 9868110 TI - [Effects of ischemic preconditioning on protection of canine donor lung]. AB - The authors examined the effects of ischemic preconditioning on lung preservation using a canine single left lung transplantation. Twelve adult mongrel dogs underwent left lung allotransplantation. Donor lungs were perfused and flushed with cold Euro-Collins solution (ECS) and stored at 4 degrees C ECS for 2 hours. Six donors were preconditioned by occuluding left lung hilum for 10 minutes and releasing for 15 minutes before flushing (Group PC); other six donors without ischemic preconditioning served as the controls (Group C). Left inferior pulmonary vein blood gas analysis, mean pulmonary artery pressure measurement and donor lung histology examination were made to evaluate the function of transplanted lung after transplantation. Oxygen tension at 2 hrs after reperfusion were significantly better in Group PC than in Group C (431 +/- 130 mmHg vs 246 +/- 66 mmHg, P < 0.05); mean pulmonary artery pressure was much lower in Group PC than in Group C after reperfusion (20.6 +/- 1.3 mmHg vs 36.9 +/- 3.1 mmHg, P < 0.01). Histological findings showed less injury in Group PC. These indicate that ischemic preconditioning combined with cold ECS perfusion is superior to cold ECS perfusion alone in canine lung preservation of 3 hours ischemia with 2 hours reperfusion. PMID- 9868111 TI - [Studies on purging leukemic cells by photosensitizer PSD-007 laser photoradiation in vitro]. AB - The sensitivity to photosensitization mediated by the hematoporphyrin photosensitizer PSD-007 of acute promyelocytic leukemic cell line (HL-60) was compared with normal human hemopoietic progenitor cells. The results showed that the leukemic cells were more sensitive. After being treated with 10 micrograms.ml 1 PSD-007 followed by 2J.cm-2 copper vapor laser light irradiation, the clonogenic leukemic (HL-60) cells were reduced 98%, but the survival rate of human granulocyte-macrophage colony-forming units (CFU-GM) was 40 +/- 8%. Mixing of normal human marrow cells with leukemic (HL-60) cells (ratio 100:1) did not interfere with elimination of tumor cells. The ultrastructure changes of HL-60 cells treated by laser photoradiation was observed under the trans-electronic microscope. The mitochondria, endoplasmic reticulum and cell membrane were involved. This means that the cell biomembrane is the main target to be attacked. It is considered that PSD-007-laser photoradiation therapy is efficient for killing leukemic cells. PMID- 9868112 TI - [The effects of deferoxamine on bovine pulmonary endothelial cell injury induced by hydrogen peroxide]. AB - The effect of deferoxamine (DFX), the ferric iron chelator, on bovine pulmonary endothelial cell (BPAEC) injury induced by hydrogen peroxide (H2O2) was examined in vitro. It was found that, when compared with unpretreated cells, H2O2-induced the release of lactate dehydrogenase and the production of thiobarbituric acid reactive substances were decreased, at the meantime the cellular activites of catalase and superoxide dismutase were maintained in DFX (2 mmol.L-1) pretreated BPAECs challenged by 1 mmol.L-1 H2O2. These data indicate that DFX provided almost complete protection against H2O2-mediated cytotoxicity. This suggests that intracellular iron may play an essential role in the endothelial cell injury mediated by H2O2. PMID- 9868113 TI - [Detection of human cytomegalovirus infection in blood donor population by the polymerase chain reaction]. AB - Samples of peripheral blood from 76 different donors were detected for the presence of human cytomegalovirus (HCMV) DNA by polymerase chain reaction (PCR). The results showed that HCMVs were present in 39 out of 76 blood donors which reached an infection positive rate of 51.3%. In addition, the sensitivity and specificity on PCR were tested, it also indicated that the sensitivity can reach the level of 5fg.microliter-1. The method is simple, rapid, highly sensitive and specificity. It can be considered as a diagnostic measure in clinical detection of HCMV. PMID- 9868114 TI - [A preliminary study on p53 gene expression and infection of human papilloma virus in laryngeal squamous cell carcinoma]. AB - Using immunohistochemical techniques with p53 monoclonal antibody DO-7 and polymerase chain reaction with type specific primers, we detected the expression of p53 of laryngeal and hypopharyngeal squamous cell carcinoma (L-HSCC) in 42 patients, tissues around tumor in 25 patients, human papilloma virus (HPV) 16/18 DNA in paraffinembedded carcinoma tissues from 13 patients with laryngeal squamous cell carcinoma (LSCC). The results showed that overexpression of p53 was detected in 54.8% (23/42) of L-GSCC and 20% (5/25) of hyperlasia epithelia, respectively. There was no correlation of p53 overexpression with clinical stages and histological grading of tumors (P > 0.05). HPV16 DNA encoding E6 protein was detected in 23.1% (3/13) LSCC tissues by PCR. The results suggest that overexpression of p53 and HPV infection are not only associated with pathogenesis of this kind of cancer but also cooperated during carcinogenesis. PMID- 9868116 TI - [Clinical study on plasma beta-endorphin levels in patients with wetness-heat zheng of the liver and gallbladder]. AB - The plasma concentrations of beta-endorphin was determinated by radioimmunoassay in 75 patients with Wetness-Heat Zheng of Liver and Gallbladder, 23 patients with Insufficient Liver Zheng, 13 patients with Insufficient Spleen Zheng and 30 healthy subjects (control group). Compared with the control group, the plasma beta-EP levels were increased in Wetness-Heat Zheng of Liver and Gallbladder and Insufficient Liver Zheng. The patients with different diseases that were diagnosed to be Wetness-Heat Zheng of Liver and Gallbladder presented high levels of plasma beta-EP. It is suggested that the plasma beta-EP is an important change in the syndrome of Wetness-Heat Zheng of Liver and Gallbladder, and may be used as a diagnostic index. PMID- 9868115 TI - [Analysis of the curative effect on ascites in liver cirrhosis by integrated traditional Chinese and Western medicine]. AB - Eighty eight patients with cirrhosis ascites were treated with regular medicine and peritoneal cavity administration or with traditional Chinese medicine and western medicine. The results were as follows: The urine volume per day was markedly increased in the group treated with traditional Chinese medicine and western medicine, the abdominal circuit decreased, and the time of ascites disappearonce significantly shortened. There was a significant difference betwee the regular group and the peritoneal cavity administration group (P < 0.01); the serum albumin/globulin ratio was remarkably increased (P < 0.05). The curative effect on ascites disappearonce in the traditional Chinese medicine and western medicine group prevailed over the other two groups (P < 0.01). PMID- 9868117 TI - [Clinical application of myocardial perfusion imaging with 99mTc-MIBI SPECT in the diagnosis of coronary artery disease]. AB - Myocardial perfusion imaging with 99mTc-MIBI was studied in 80 patients with coronary artery disease (CAD), 29 cases of normal controls and 37 cases of noncoronary heart diseases. The perfusion imaging was analysed both by qualitative and quantitative methods. The results revealed that the diagnostic sensitivity, specificity and accuracy for CAD were 91.5%, 83.8%, and 88.5% respectively by qualitativ method, and were 88.1%, 94.6% and 90.6% respectively by quantitative method. The diagnostic sensitivity of electrocardiogram (ECG) was 69.5%. Fourteen cases with the narrowing of coronary artery > 50% shown by coronary angiography also had abnormal myocardial perfusion imaging. It suggests that analysis of perfusion imaging by quantitative method in detecting CAD is superior than qualitative method and ECG. PMID- 9868118 TI - [Effects of inhalated isosorbide dinitrate on endothelin and nitric oxide and its therapeutic efficacy for experimental guinea-pigs with asthma]. AB - To probe into the correlation between the changes of endothelin and nitric oxide in bronchial alveolar lavage fluid (BALF) and the treatment of inhaling isosorbide dinitrate on experimental guinea pigs with asthma, we divided 25 guinea pigs randomly into three groups (Control group, Asthma group, Isosorbide dinitrate group), and induced some subjects by inhalation of aerosolized ovalbumin as the model of asthma to investigate the endothelin (ET) and nitric oxide (NO) level in BALF as well as the pathological change of the respiratory tract. We found that the level of endothelin and nitric oxide in BALF were all elevated in experimental guinea pigs with asthma, but the degree of increase of NO in BALF was lower than that of ET. The NO and the ET levels in BALF were negative correlation (r = -0.8050, P < 0.01). After having been treated with aerosolized isosorbide dinitrate, the NO level in BALF was significantly elevated and the ET level in BALF was decreased in comparison with the asthma group (P < 0.001, respectively). Moreover, the eosinophile infiltration, exoserosis, and epithalaxia of the respiratory tract of the guinea-pigs were all lightened. The results suggest that inhaled isosoratory dinitrate could be used for treating asthma, and its mechanism may be elevation of NO level and decrease of ET level in BALF. PMID- 9868119 TI - [Detection of the level of serum IgG antibody to hepatitis E virus]. AB - The levels of serum IgG antibody to hepatitis E virus (anti-HEV-IgG) was detected in serum specimens from 50 patients with acute sporadic hepatitis E and 20 healthy donors who were positive for anti-HEV-IgG by ELISA titration. The results showed that the level of anti-HEV-IgG in acute hepatitis E patients was significantly higher than that in healthy donors and the average titer is 1:121 and 1:22, respectively (P < 0.01). So acute viral hepatitis E should be diagnozed when the titer of anti-HEV-IgG was over 1:40. PMID- 9868120 TI - [Differentiation of benign and malignant palpable breast masses using 99mTc-MIBI imaging]. AB - Forty-two patients with palpable breast masses were examined by 99mTc-MIBI imaging. All the results were compared with ultra-red scanning and B ultrasongraphy. By pathological examination 26 out of 42 cases had breast carcinoma, 16 patients benign lesions. The sensitivity of 99mTc-MIBI imaging was 92.3%, the specificity was 96.0% and the accuracy 92.8%, which were higher than those of ultra-red scanning and B-ultrasound (P < 0.01). The authors consider that 99mTc-MIBI breast imaging is useful in distinguishing malignancies from benign breast masses. PMID- 9868121 TI - [A study of anaerobic infection in maxillofacial region]. AB - To analyse the anaerobic infection of maxillofacial surgery and estimate the efficacy of antianaerobic therapy, 45 patients were divided into two groups, tinidazole group and metronidazole group. Bacterial culture was positive before treating in all cases. There were Bacillus Melaniogenicus, Veillonella, Peptococcus and Peptostreptococus, etc. There was excellent efficacy in the treatment of maxillofacial anaerobic infection by tinidazole intravenously. After treatment, the result of bacterial examination was negative. The healing rate was 96.4% in 28 cases which used tinidazol, but 82.4% in control group which used metronidazole. The value of white blood cell and the function of liver and kidney pro- and post-treatment were not significantly different (P < 0.05) by comparision. PMID- 9868122 TI - [Effects of perindopril on hypertension, serum angiotensin-I-converting enzyme activity and urine level of N-acetyl-beta-D-glucosaminidase]. AB - Perindopril 4 mg once daily was given to 40 hypertensives for 4 weeks. The results showed that systolic and diastolic blood presure were decreased by 3.2 kPa (22.2 +/- 2.21-19.0 +/- 1.92 kPa) and 1.87 kPa (13.4 +/- 1.21-11.5 +/- 1.27 kPa), respectively. The total effective rate was 80%. Serum Angiotensin-I converting enzyme activity in 30 patients was significantly decreased from 58.5 +/- 29.5 U to 18.2 +/- 16.2 U (P < 0.01). The urine level of N-acetyl-beta-D glucosaminidase (NAG) in 27 patients significantly decreased from a prior level of 13.66 +/- 7.81 U.gCr-1 to 10.12 +/- 5.57 U.gCr-1 (P < 0.01). The side effects of perindopril were cough (7.5%), constipation (10%), dizziness (7.5%), flatulence (7.5%) and diarrhea (5%). We conclude that perindopril is a potent antihypertensive drug with significant prevention on hypertension-induced early renal damage. PMID- 9868123 TI - [Effects of danshen on solute transport by peritoneal dialysis]. AB - Danshen injection was added in peritoneal dialysate with concentration of 6/1000. The D/P value of BUN, Cr and total protein and the D/Do value of glucose and the drained volume were observed. The results showed that the D/P value of BUN and Cr increased from the 2nd day of Danshen addition; the D/Do value of glucose increased from the 1st day of Danshen addition and then came to the original value from the 2nd day of Danshen addition; the D/P value of protein increased continuously from the 1st day of Danshen addition; the drained volume had no change. PMID- 9868124 TI - [Image analysis of tumor cells from common intracranial neoplasms on frozen and paraffined slices]. AB - Image analysis technique was used to measure four parameters [the form factor (FF), area, perimeter and diameter] of nuclei in 76 cases of common intracranial tumors on forzen and paraffined slices. The results showed that the area, perimeter, diameter of nuclei on frozen slices were much larger than those on paraffined slices while the FF showed no significant difference between two kinds of slices (P > 0.05). The parameters of the area, perimeter, and diameter of nuclei on two kinds of slices became larger and the parameters of FF became smaller as the grade of astrocytoma ascended. It is considered that the results of present study will be helpful to the diagnosis of common intracranial tumors on frozen slices. PMID- 9868125 TI - [Clinical valuation color Doppler echocardiography for diagnosis of coronary artery fistula]. AB - The purpose of this study was to investigate the advantages of color Doppler echocardiography in the diagnosis of congenital coronary artery fistula. Twenty one patients with coronary artery fistula ranging in age from 3 to 28 years underwent color Doppler echocardiographic study 1991.4-1996.8. When a coronary artery fistula was found, the origin, the course and the drainage site were carefully searched for to make a precise diagnosis. Color Doppler echocardiographic diagnosis of all patients were confirmed by cardiac operation on cardioangiography. The results indicate that color Doppler echocardiography can provide a panoramic view of coronary artery and the fistulous vessel with precise definition of the origin, the course of draingae site of the fistula. Therefore, it is the noninvasive method of first choice to diagnose congenital coronary artery fistula. PMID- 9868126 TI - [The preventing function of garlic on experimental oral precancer and its effect on natural killer cells, T-lymphocytes and interleukin-2]. AB - In order to study the effect and mechanism of garlic on preventing oral precancer, we divided randomly 32 Wistar rats into two groups. The garlic group was painted with garlic solution on the hard palatal mucosae. The control group was applied with distilled water that is equal in quantity. Then, chemical carinogen 4-nitroquinoline 1-oxide (4NQO) was painted on the same sites for both groups, three times weekly. Eight rats were randomly killed in the 10th, 13th week. The hard palatal mucosae were examined with light microscope. Meanwhile, lymphocytes were isolated from the rat spleens. The activation of natural killer (NK) cells and T-lymphocytes, and level of interleukin-2 were determined by radioimmunoassay. The results revealed that garlic effectively prevented oral precancer induced by 4NQO. This effect may be related to the following factors that garlic can improve the activation of NK cells, the function of T lymphocytes, and the level of IL-2. PMID- 9868128 TI - [Color Doppler echocardiography in the diagnosis for ruptured aneurysm of sinus of Valsalva]. AB - Twenty-five cases with ruptured aneurysm of sinus of Valsava were detected by color Doppler sonography and twenty-four cases undenwent surgical repair. In twenty-three cases, the sonographic findings of the site of the aneurysm and the chambers into which aneurysm ruptured were coincident with the surgical findings. The accuracy was 95.8%. The size of the aneurysm and the diameter of the ruptured site detected by ultrasound were correlated well with surgical findings. The associated lesions obtained by ultrasound were completely identical with surgical findings. Color Doppler flowing imaging has special value in diagnosing the ruptured aneurysm of sinus of Valsalva. PMID- 9868127 TI - [The clinical effect after locally using mitomycin C during trabeculectomy in glaucoma]. AB - To improve the function of filtering blebs after trabeculectomy, we divided randomly 44 patients (60 eyes) with moderate or late glaucoma into mitomycin C and control groups (30 eyes per group). Two small surgical sponges were soaked with 0.2 mg.ml-1 mitomycin C solution. One sponge was put on the top of the superficial scleral flap, the other was placed underneath the scleral flap and contacted with the bed of the scleral flap. After 5 minutes the wound was then irrigated with normal saline. All patients were followed up from 1 to 12 months (7.9 +/- 4.0 months). We found that: (1) functional filtering blebs were 92.9% in the mitomycin C group and 55.2% in the control group (P < 0.01), (2) the decreasing percentage of intraocular pressure (IOP) in the mitomycin C group was much more than that in the control group (P < 0.05), (3) the successful rates of the operation in MMC group were higher than that of the control group, and the visual field was effectively maintained in MMC group. PMID- 9868129 TI - [Comparison between intraarterial pulsegraphy and thermodilution method for determination of cardiac output during anesthesia and operation in patients undergoing cardiac valve replacement]. AB - Comparison of intraarterial pulsegraphy and standard thermodilution methods for determination of cardiac output (CO) in 14 patients with poor cardiac function following valve replacement operation was performed. The CO, cardiac index (CI) were moderate to high correlated between the two methods before and after valve replacement respectively. The study indicates that intraarterial pulsegraphy provides a continous method of determining CO in patients with poor cardiac function during operation. PMID- 9868130 TI - [Relationship between urinary albumin excretion and blood vessel lesion in non insulin-dependent diabetics]. AB - To search the relationships between urinary albumin excretion and blood vessel lesion in non-insulin-dependent diabetics, we compared the differences of blood lipid, blood pressure, coronary heart disease and retina disease between 58 patients. The group with microalbuminuria had significantly lower concentration of apolipoprotein A, and apolipoprotein A1/apolipoprotein B100 than that of the group with normal albuminuria, but there was no significant change in concentrations of trigly cerides, total cholesterol, high density lipoprotein, and apolipoprotein B100. Coronary heart disease and retina disease were significantly increased in group with microalbuminuria. There was negative correlation with apolipoprotein A1 and value of apolipoprotein A1/apolipoprotein B100, but positive correlation with systolic blood pressure by linear regression analysis for urinary albumin excretion. PMID- 9868131 TI - [Clinical analysis of spontaneous perforation of abdominal viscera in patients with severe viral hepatitis]. AB - Eight out of 246 patients with severe viral hepatitis were complicated with spontaneous abdominal viscera perforation. The eight patients all had serious digestive symptoms, orange green colour in skin, swollen abdomen with ascites, abdominal pain. All cases were diagnosed by liver function examination, colour ultrasonography, x-ray examination, abdominal puncture and one case by operation. Among these 8 cases, 3 cases had perfornation of stomach, 3 cases had perforation of gall bladder and 2 cases had perforation of small intestine. Though emergancy treatment had been applied, only one case was saved. PMID- 9868132 TI - [Application of T1-weighted images by contrast-enhanced frequency-selective presaturation in magnetic resonance imaging]. AB - In this study contrast-enhanced frequency-selective presaturation T1-weighted images and contrast-enhanced spin echo T1-weighted images in 32 cases were detected and analysed comparatively regarding enhancing lesions and the signal intensity of enhancing lesions. The results showed that in the signal intensity of enhancing lesion, the contrast enhanced frequency-selective presaturation T1 weighted images showing high sign were greater than contrast-enhanced spin echo T1-weighted images. The former was better than the later in accurate evaluation of the margin, size and the differential diagnosis of enhancing lesion, especially in the regions containing more fat and bone marrow. PMID- 9868133 TI - [The effect of root canal irrigation and variance of apical foramen pressure by use of casing aspiration needle system in vitro]. AB - The authors introduced casing aspiration needle system (CANS) to irrigating root canal. The effect of root irrigation on removing canal debris and the variance of apical foramen pressure on extracted teeth were compared between the conventional irrigation and CANS irrigation. The results indicated that the effect of root canal irrigation by CANS was much better than the conventional method (P < 0.01). CANS irrigation didn't produce pressure to apical foramen during root canal irrigation. However, in the conventional method, with irrigation needle inserted deeper and deeper in root cancal the pressure of apical foramen gets higher and higher. PMID- 9868134 TI - [Blood levels of calcitonin gene related peptide, atrial natriuretic factor and aldosterone in liver-blood deficiency syndrome]. PMID- 9868135 TI - [Role of thiamin in the prevention of erythrocyte damage by lead]. PMID- 9868136 TI - [Scavenging effects of sodium selenite on hydroxyl radicals]. PMID- 9868137 TI - [Assay of serum beta 2-microglobulin in patients with malignant hematologic diseases and its clinical significance]. PMID- 9868138 TI - [Cell-mediated and humoral immunity in human rabies]. PMID- 9868140 TI - [A case of big pseudotumor of the biorbit]. PMID- 9868139 TI - [Demodex infection in university and middle school students]. PMID- 9868141 TI - [Delayed encephalopathy in childhood caused by anthelmintics]. PMID- 9868142 TI - Antiplatelet aggregation constituents from Annona purpurea. AB - Bioactivity-directed fractionation led to the isolation of 19 compounds, including three oxoaporphines, oxopurpureine (5), oxonuciferine (6), and oxoglaucine (7); three aporphines, (+)-predicentrine (8), (-)-glaucine (9), and thalbaicalidine (10); one aporphine sensu stricto, N-formyl-purpureine (11); one proaporphine, glaziovine; one phenanthrene, thalicpureine (12); two 6a,7 dehydroaporphines, dehydrolirinidine (13) and 7-hydroxy-dehydroglaucine (14); four flavonoids, quercetin-3-O-rhamnoside, kaempferol-3-O-rhamnoside, isorhamnetin-3-O-rhamnoside, and tanarixetin-3-O-rhamnoside; one purine, adenine; one lactam amide, squamolone; and two steroids, beta-sitosterol and beta sitosterol-beta-D-glucoside from the MeOH extract of the leaves of Formosan Annona purpurea. Among them, 11-14 were characterized as new compounds and alkaloids, 5-8, 10, and 12-14 exhibited significant antiplatelet aggregation activity. PMID- 9868143 TI - Minor cassane diterpenoids of caesalpinia bonduc AB - Seven cassane diterpenes, including the known caesaldekarin A, were isolated from the roots of Caesalpinia bonduc, collected in Barbados. The 1H and 13C NMR spectra of all seven compounds were completely assigned by using a combination of 2D NMR experiments, which included 1H-1H COSY, HSQC, HMQC, and HMBC sequences. PMID- 9868144 TI - Tetrahydropyran monoterpenes from plocamium cartilagineum and pantoneura plocamioides AB - Four new marine monoterpenes have been isolated from two species of marine red algae, Plocamium cartilagineum and Pantoneura plocamioides. The structures and relative stereochemistry of these compounds were determined on the basis of spectroscopic evidence and suggest a relationship between P. cartilagineum and P. plocamioides. PMID- 9868145 TI - Cycloartane triterpene glycosides from the roots of Astragalus brachypterus and Astragalus microcephalus. AB - Three new cycloartane-type triterpene glycosides, brachyosides A (1), B (3), and C (2), from the roots of Astragalus brachypterus and one new glycoside, cyclocephaloside II (4), from the roots of Astragalusmicrocephalus have been isolated together with five known saponins, astragalosides I, II, and IV, cyclocanthoside E, and cycloastragenol. The structures of the new compounds were established as 3-O-[beta-D-xylopyranosyl(1-->3)-beta-D-xylopyranosyl-6-O-beta-D gluc opyranosyl-3beta,6alpha,16beta,24(S),25-pentahydrox ycycloartane (1), 3-O beta-D-xylopyranosyl-6-O-beta-D-glucopyranosyl-24-O-beta-D-glucop yranosyl 3beta,6alpha,16beta,24(S),25-pentahydroxyc ycloartane (2), 20(R),24(S)-epoxy-6-O beta-D-glucopyranosyl-3beta,6alpha,16beta , 25-tetrahydroxycycloartane (3), and 20(R), 24(S)-epoxy-3-O-(4'-O-acetyl)-beta-D-xylopyranosyl-6-O-beta-D-glucopy ranosyl-3beta,6alpha,16beta,25-tetrahydroxycycloart ane (4). For the structure elucidations, 1D- and 2D-NMR experiments and FABMS were used. PMID- 9868146 TI - Grayanane diterpenoids from pieris formosa AB - Two new diterpenoids, pierisformoside A (1) and pierisformosin D (2), and two known diterpenoids, asebotoxins VIII (3) and V (4), were isolated from leaves of Pieris formosa. Their structures were elucidated on the basis of spectral analysis, including 1H-1H COSY, 13C-1H COSY, HMBC, and NOESY experiments. PMID- 9868147 TI - Isolation and characterization of novel endogenous digitalis-like factors in the ovary of the giant toad, Bufo marinus. AB - We have previously described the structures of four novel unconjugated bufadienolides in the ovary of the toad, Bufo marinus. In this study, we report the separation and characterization of three novel bufadienolide conjugates. These compounds were purified by HPLC, and their structures were determined to be 11alpha, 19-dihydroxytelocinobufagin-3-(12-hydroxydodecanoic acid) ester, 11alpha,19-dihydroxytelocinobufagin-3-(14-hydroxy-7-tetra decenoic acid) ester, and 11alpha, 19-dihydroxytelocinobufagin-3-(14-hydroxytetradecanoic acid) ester on the basis of NMR and MS data. Numerous dicarboxylic acid esters of bufadienolides have previously been described, but the three bufadienolide conjugates described in this report differ from previously described esters in that they contain hydroxylated monocarboxylic acids. The function of these three conjugates is not known but they are, like bufotoxins, potent inhibitors of Na+, K+-ATPase and may play a developmental role in the differentiation of toad oocytes. PMID- 9868148 TI - Bisamides, lignans, triterpenes, and insecticidal Cyclopenta[b]benzofurans from Aglaia species. AB - Twelve new derivatives of bisamides [piriferinol (5), edulimide (7)], lignans [lariciresinol acetat (10)], triterpenes [4-bis-norcycloartane-type triterpenes (11a, 12a+b, 13a+b)], cyclopenta[b]benzofurans [pannellin (15), pannellin 1-O acetate (16), 3'-methoxypannellin (17)], and an aromatic butyrolactone [aglalactone (18)] were isolated and identified from lipophilic leaf, stem, and root extracts of Aglaia spp. elaeagnoidea, edulis, grandis, silvestris, and tomentosa. Two cyclopenta[b]benzofurans (flavaglines), pannellin (15) and pannellin 1-O-acetate (16), displayed very strong insecticidal activity against Spodoptera littoralis and Lymantria dispar, comparable with the activity of azadirachtin. PMID- 9868149 TI - Phycomysterols and other sterols from the fungus Phycomyces blakesleeanus. AB - In the search for novel bioactive products from filamentous fungi, sterols and triterpenoids found in Phycomyces blakesleeanus were analyzed using semipreparative HPLC, GC-MS, and NMR techniques. Structures proposed for the three new compounds identified, phycomysterol A (1), phycomysterol B (2), and neoergosterol (3), were confirmed by chemical synthesis. Phycomysterols possess a new natural 19-norergostane skeleton with an aromatic B ring. Phycomysterol A showed anti-HIV activity. PMID- 9868150 TI - Use of on-flow LC/1H NMR for the study of an antioxidant fraction from Orophea enneandra and isolation of a polyacetylene, lignans, and a tocopherol derivative. AB - The CH2Cl2 extract of the leaves of Orophea enneandra displayed antifungal, antioxidant, and radical scavenging properties in bioautographic TLC assays. To obtain rapid information on the active compounds, on-flow LC/1H NMR and LC/UV/MS analyses of the antioxidant fraction were performed. The on-line information led rapidly to the partial identification of three closely related lignans, one tocopherol derivative, and one polyacetylene. This approach necessitated, however, large quantities to be injected to obtain satisfactory on-flow LC/1H NMR spectra, and isolation of the compounds was necessary to obtain complete NMR data. These compounds were isolated and identified as (-)-phylligenin (1), (-) eudesmin (2), (-)-epieudesmin (3), polycerasoidol (4), and oropheic acid (5), a new polyacetylene. Their activities against the 2, 2-diphenyl-1-picrylhydrazyl radical and the fungus Cladosporium cucumerinum were investigated. This paper indicates the possibilities and limits of on-flow LC/1H NMR in this type of study. PMID- 9868151 TI - Chemistry of puupehenone: 1,6-conjugate addition to its quinone-methide system. AB - The marine natural product puupehenone (1), isolated in good yields from sponges of the genus Hyrtios, has been shown to undergo stereospecific 1,6-conjugate addition to its quinone-methide system. Several nucleophilic agents such as hydrogen cyanide, Grignard reagents, and nitroalkanes were studied, producing structurally diverse compounds. This lead optimization study was initiated due to the bioactivity of puupehenone and its natural analogues, which includes numerous previous reports of potential anticancer and antiinfective activity. PMID- 9868153 TI - New xenia diterpenes isolated from the soft coral, xenia florida AB - Seven new xenia diterpenes have been isolated from the soft coral, Xenia florida. Four of them possess a bicyclic [4.3.1] ring system. The three others, which contained a cyclononane skeleton with an epoxide, seem to be precursors for diterpenes with a bicyclic [4.3. 1] ring system. PMID- 9868152 TI - Cytotoxic geranyl stilbenes from Macaranga schweinfurthii. AB - Three novel geranyl stilbenes, schweinfurthins A, B, and C (1, 2, and 3), were isolated from the Cameroonian plant Macaranga schweinfurthii (Euphorbiaceae) and their structures determined by NMR and mass spectral methods. The cytotoxicity profile of the schweinfurthins tested in the NCI 60-cell screen was similar to that of the stelletins and cephalostatins, suggesting that these structurally diverse natural products may share similar mechanisms of cytotoxicity. PMID- 9868154 TI - Phytoalexins from hairy roots of Hyoscyamus albus treated with methyl jasmonate. AB - The treatment of hairy roots of Hyoscyamus albus with copper sulfate (Cu2+) and methyl jasmonate (JAMe) produced several phytoalexins having the vetispyrane skeleton. Lubimin and solavetivone were isolated after treatment with Cu2+. Seven sesquiterpenoid phytoalexins were isolated from the culture medium after treatment with JAMe, including lubimin, solavetivone, 3-hydroxysolavetivone and four new compounds (1-4). Structures of the new compounds were elucidated to be (3R,4S,5R,7S,9R)-3-hydroxy-9-tigloyloxysolavetivone (1), (3R,4S,5R,7S,9R)-3 hydroxy-9-(3-methylbutenoyloxy)-solavetivone (2), (3R,4S,5R,7S,9R)-3-hydroxy-9 isobutanoyloxysolavetivone (3); and (3R,4S,5R,7S,9R)-3,9-dihydroxysolavetivone (4). The induction pattern of phytoalexins in hairy roots treated with JAMe was different in those treated with Cu2+, and co-treatment with JAMe and Cu2+ gave only solavetivone. PMID- 9868155 TI - New sesquiterpenes from Crossopetalum tonduzii. AB - Five new sesquiterpenes (1-5) with a dihydro-beta-agarofuran skeleton were isolated from Crossopetalum tonduzii. Their structures were elucidated by means of 1H and 13C NMR spectroscopic studies, including homonuclear and heteronuclear correlation experiments (COSY, ROESY, HMQC, and HMBC). The absolute configurations of 1 and 2 were determined by CD studies and chemical correlation. Compounds 1-3 were assayed against Spodoptera littoralis in an election test and showed low insect-antifeedant activity. PMID- 9868156 TI - New diterpene isonitriles from the sponge Phakellia pulcherrima. AB - Seven new diterpene isonitriles and isothiocyanates were isolated from the sponge Phakellia pulcherrima along with eight known ones. Six of the new compounds, 9 14, and the eight known ones, 1-8, belong to the kalihinol family of diterpenes. Structures were determined from spectroscopic data. This is the first report of diterpenes from sponges of this genus. All of the other kalihinol diterpenoids have been isolated from the sponge Acanthella cavernosa collected in diverse locations. PMID- 9868157 TI - Isolation and unambiguous synthesis of cryptolepinone: An oxidation artifact of cryptolepine AB - Cryptolepinone (3) was isolated as an artifact of extraction from Cryptolepis sanguinolenta. Previously, this compound had been identified as the natural products hydroxycryptolepine (2) and 3. Synthesis via an unambiguous pathway has confirmed the structure of cryptolepinone. Spectroscopic studies in various solvents have shown that the natural product artifact or its synthetic equivalent can exist in the keto (cryptolepinone) or enol (hydroxycryptolepine) form. PMID- 9868158 TI - Cytotoxicity of curcuminoids and some novel compounds from Curcuma zedoaria. AB - Bioassay-directed fractionation of an EtOH extract of Curcuma zedoaria led to isolation of an active curcuminoid, which was identified as demethoxycurcumin (2) by comparison of its 1H and 13C NMR spectra with literature data and by direct comparison with synthetic material. Curcumin (1) and bisdemethoxycurcumin (3) were also obtained. Curcuminoids (1-3) were synthesized and demonstrated to be cytotoxic against human ovarian cancer OVCAR-3 cells. The observed CD50 values of 1, 2, and 3 were 4.4, 3.8, and 3.1 microg/mL, respectively. Three additional novel compounds, 3, 7-dimethylindan-5-carboxylic acid (4), curcolonol (5), and guaidiol (6), were also isolated from the EtOH extract. The structures and relative stereochemistry of 4-6 were determined by spectroscopic methods and X ray crystallographic analysis. PMID- 9868159 TI - Novel cytotoxic acylated oligorhamnosides from Mezzettia leptopoda. AB - Activity-guided fractionation of a stem extract of Mezzettia leptopoda using human oral epidermoid carcinoma (KB) cells led to the isolation of seven highly acylated oligorhamnosides. Four of these constituents are novel, namely, n-octyl 2-O-acetyl-alpha-L-rhamnopyranosyl-(1-->3)-2, 4-di-O-acetyl-alpha-L rhamnopyranosyl-(1-->3)-4-O-hexanoyl-alpha-L-rh amnopyranoside (mezzettiaside 8) (1); n-octyl 2, 3-di-O-acetyl-alpha-L-rhamnopyranosyl-(1-->3)-4-O-hexanoyl-alpha L-rh amnopyranoside (mezzettiaside 9) (2); n-octyl 2, 4-di-O-acetyl-alpha-L rhamnopyranosyl-(1-->3)-4-O-hexanoyl-alpha-L-rh amnopyranoside (mezzettiaside 10) (3); and n-octyl 2,3, 4-tri-O-acetyl-alpha-L-rhamnopyranosyl-(1-->3)-4-O-hexanoyl alpha-L-r hamnopyranoside (mezzettiaside 11) (4). Three known compounds were identified as mezzettiasides 2 (5), 3 (6), and 4 (7), respectively, previously isolated from this same plant. The structures of novel compounds 1-4 were determined by spectroscopic methods. All the isolates were evaluated against a panel of human cancer cell lines in this study, and compounds 1-2 and 4-7 were found to be weakly cytotoxic toward KB and/or human colon and lung cancer cell lines. PMID- 9868160 TI - New cyclic peroxides from the Philippine sponge Plakinastrella sp. AB - Three new cyclic peroxides 5-7 and a new carboxylic acid ester 8 were isolated as minor metabolites from the hexane extract of a Plakinastrella species from the Philippines. The structures of compounds 5-8 were elucidated by interpretation of spectral data and by chemical interconversion, and the absolute stereochemistry of peroxide 6 was determined by application of Mosher's method to a derivative. Although the major compounds in the sponge showed activity against Candida albicans prior to decomposition, the minor metabolites 5-8 are essentially inactive. PMID- 9868161 TI - Facile total synthesis and antimicrobial activity of the marine fatty acids (Z)-2 methoxy-5-hexadecenoic acid and (Z)-2-methoxy-6-hexadecenoic acid. AB - The total synthesis of the naturally occurring (Z)-2-methoxy-5-hexadecenoic acid and (Z)-2-methoxy-6-hexadecenoic acid was accomplished using as a key step Mukaiyama's trimethylsilyl cyanide addition to 4- and 5-pentadecenal, respectively. These syntheses further confirm the structures of the natural marine fatty acids and corroborate their cis double-bond stereochemistry. The title compounds were antimicrobial against the Gram-positive bacteria Staphylococcus aureus (MIC 0.35 micromol/mL) and Streptococcus faecalis (MIC 0.35 micromol/mL). PMID- 9868162 TI - New cyclic peptides from the ascidian Lissoclinum patella. AB - Four new cyclic peptides, patellamide G (2) and ulithiacyclamides E-G (3-5), along with the known patellamides A-C (6-8) and ulithiacyclamide B (9), were isolated from the ascidian Lissoclinum patella collected in Pohnpei, Federated States of Micronesia. The planar structures of these peptides were determined from 1D and 2D 1H and 13C NMR spectra. The absolute stereochemistries of the amino acid units, except for cysteine, were assigned by chiral GC analysis of N(O)-trifluoroacetyl isopropyl ester derivatives of amino acids obtained by acid hydrolysis of the intact and ozonized peptides. The structures of ulithiacyclamides E-G (3-5) were confirmed by chemical conversion. Patellamides B (7) and C (8) exhibited in vitro modulation of multidrug resistance in CEM/VBL100 cells. PMID- 9868163 TI - A new prenylated flavonol from the roots of Sophora flavescens. AB - A new prenylated flavonol, sophoflavescenol (1), together with five known flavonoids, kurarinol, kushenol K, kushenol H, trifolirhizin, and kuraidin, were isolated from the roots of Sophora flavescens. The structure of 1 was determined by spectroscopic analysis. Among the five known flavonoids, kurarinol, kushenol K, and kushenol H showed weak antiviral activity against Herpes simplex virus types I and II. PMID- 9868164 TI - Beiwudine, a norditerpenoid alkaloid from aconitum kusnezoffii AB - A novel norditerpenoid alkaloid, beiwudine (1), was isolated from the roots of Aconitum kusnezoffii. Its structure was established on the basis of chemical and NMR spectral studies. PMID- 9868165 TI - Two new sulfated saponins from the roots of Gypsophila bermejoi. AB - Two new sulfated saponins (1 and 2) were isolated from a butanol-soluble extract of the roots of Gypsophila bermejoi and were identified by a combination of chemical degradation and spectral methods as the 3beta-sulfate ester of gypsogenin 28-O-beta-D-glucopyranosyl(1-->2)-[beta-D-glucopyranosyl(1-->6)]-beta D-glucopyranoside (1), and the 23-sulfate ester of hederagenin 28-O-beta-D glucopyranosyl(1-->2)-[beta-D-glucopyranosyl(1-->6)]-beta -D-glucopyranoside (2), respectively. Plants of the genus Gypsophila (Caryophyllaceae) are important industrially because of the capacity of their saponin constituents to behave like natural detergents.1 Saponins from this genus are based on oleanolic acid (3 hydroxyolean-12-en-28-oic acid), with gypsogenin being the main pentacyclic triterpenoidal aglycon found. In general, this aglycon is substituted at the C-3 hydroxyl and/or C-28 carboxylic acid groups by saccharide units.2 PMID- 9868166 TI - A new indole derivative from the red alga Chondria atropurpurea. Isolation, structure determination, and anthelmintic activity. AB - Chondriamide C (3), a new bis(indole) amide, was isolated from the red alga Chondria atropurpurea, and its structure was established from spectroscopic data and chemical transformations. A new natural product, 3-indoleacrylamide (4), and the previously described chondriamides A and B (1, 2) and 3-indoleacrylic acid (5) were also isolated. The anthelmintic activities of compounds 1, 3, 4, and 6 (the O,N1,N1'-trimethyl derivative of compound 2) against Nippostrongylus brasiliensis in vitro were evaluated. PMID- 9868167 TI - (+)-Bulbocapnine-beta-N-oxide from glaucium fimbrilligerum AB - A new aporphine alkaloid, (+)-bulbocapnine-beta-N-oxide (1), was isolated from Glaucium fimbrilligerum. Its structure and the stereochemistry at the N-oxide center were determined by spectroscopic methods and confirmed by synthesis. PMID- 9868168 TI - Arugosin F: a new antifungal metabolite from the coprophilous fungus Ascodesmis sphaerospora. AB - Chemical studies of the coprophilous fungus Ascodesmis sphaerospora (JS 247) have led to the isolation of arugosin F (1), a new antifungal and antibacterial metabolite. The structure was determined based on NMR and MS data and on comparison with data for known members of the arugosin class. A known xanthone (2) was also isolated. PMID- 9868169 TI - Clavaric acid: a triterpenoid inhibitor of farnesyl-protein transferase from Clavariadelphus truncatus. AB - Farnesyl-protein transferase (FPTase) catalyses the specific transfer of farnesyl to Ras-peptides that is essential for oncogenic activity in oncogene-mediated tumors. Specific inhibition of FPTase activity has been shown to reduce tumor development in nude mice challenged with oncogenic forms of ras, thereby establishing FPTase as a viable therapeutic target. Our continued efforts to discover inhibitors of FPTase has led to the discovery of a triterpenoidal inhibitor, clavaric acid (1). This compound inhibits rHFPTase with an IC50 value of 1.3 microM. Structure elucidation, structure modifications, and biological activity of clavaric acid are herein described. PMID- 9868170 TI - Deoxynortrichoharzin, a new polyketide from the saltwater culture of a sponge derived Paecilomyces fungus. AB - The saltwater culture of a Paecilomyces cf. javanica isolated from the marine sponge Jaspis cf. coriacea has yielded a new polyketide, deoxynortrichoharzin (1), and two known diketopiperazines (2 and 3). PMID- 9868171 TI - A new meroditerpenoid dimer from an undescribed philippine marine sponge of the genus strongylophora PMID- 9868172 TI - Iron regulatory proteins, iron responsive elements and iron homeostasis. AB - The discovery of iron regulatory proteins (IRPs) has provided a molecular framework from which to more fully understand the coordinate regulation of vertebrate iron metabolism. IRPs bind to iron responsive elements (IREs) in specific mRNAs and regulate their utilization. The targets of IRP action now appear to extend beyond proteins that function in the storage (ferritin) or cellular uptake (transferrin receptor) of iron to include those involved in other aspects of iron metabolism as well as in the tricarboxylic acid cycle. To date, it appears that IRPs modulate the utilization of six mammalian mRNAs. Current studies are aimed at defining the mechanisms responsible for the hierarchical regulation of these mRNAs by IRPs. In addition, much interest continues to focus on the signaling pathways through which IRP function is regulated. Multiple factors modulate the RNA binding activity of IRP1 and/or IRP2 including iron, nitric oxide, phosphorylation by protein kinase C, oxidative stress and hypoxia/reoxygenation. Because IRPs are key modulators of the uptake and metabolic fate of iron in cells, they are focal points for the modulation of cellular iron homeostasis in response to a variety of agents and circumstances. PMID- 9868174 TI - Grape juice but not orange or grapefruit juice inhibits platelet activity in dogs and monkeys. AB - Platelet aggregation (PA) contributes to both the development of atherosclerosis and acute platelet thrombus formation (APTF) followed by embolization producing cyclic flow reductions (CFR) in stenosed and damaged dog and human coronary arteries. In seven anesthetized dogs with coronary stenosis and medial damage, CFR occurred at 7 +/- 3/30 min and were abolished 127 +/- 18 min after gastric administration of 10 mL of purple grape juice/kg. Collagen-induced ex vivo whole blood PA decreased by 49 +/- 9% after the abolishment of CFR with grape juice. Ten mL of orange juice/kg (n = 5) and 10 mL of grapefruit juice/kg (n = 5) had no significant effect on the frequency of the CFR or on ex vivo PA. In vitro studies have suggested that flavonoids bind to platelet cell membranes and thus may have an accumulative or tissue-loading effect over time. To test this we fed 5 mL of grape juice/kg to 5 cynomologous monkeys for 7 d. Collagen-induced ex vivo PA decreased by 41 +/- 17% compared to control (pre-reatment) after 7 d of feeding. In the same 5 monkeys, neither 5 mL of orange juice/kg nor 5 mL of grapefruit juice/kg given orally for 7 d produced any significant change in PA. Grape juice contains the flavonoids quercetin, kaempferol and myricetin, which are known inhibitors of PA in vitro. Orange juice and grapefruit juice, while containing less quercetin than grape juice, primarily contain the flavonoids naringin, luteolin and apigenin glucoside. The flavonoids in grapes were shown in vitro to be good inhibitors of PA, whereas the flavonoids in oranges and grapefruit to be poor inhibitors of PA. The consumption of grape juice, containing these inhibitors of PA, may have some of the protection offered by red wine against the development of coronary artery disease (CAD) and acute occlusive thrombosis, whereas orange juice or grapefruit juice may be ineffective. Thus, grape juice may be a useful alternative dietary supplement to red wine without the concomitant alcohol intake. PMID- 9868173 TI - Phenotypic consequences of a nonsense mutation in the leptin receptor gene (fak) in obese spontaneously hypertensive Koletsky rats (SHROB). AB - The genetically obese Koletsky rat (SHROB, fak) has a novel point mutation of the leptin receptor at amino acid +763, resulting in a premature stop codon in the leptin receptor extracellular domain. This implies that all leptin receptor isoforms should be absent in this model. We examined the phenotypic consequences of this mutation on leptin and leptin receptor mRNA in hypothalamus and peripheral tissues from SHROB and their lean SHR littermates. Despite the mutation, mRNA for both the long (ObRa) and the short (ObRb) form were expressed at comparable levels in SHROB and SHR in brain and throughout peripheral tissues. Adipose tissue mRNA for leptin was two to three times greater in SHROB compared to SHR (P < 0.01), while circulating leptin concentration was 170 times greater than SHR littermates (P < 0.01), suggesting extreme leptin resistance in SHROB. Leptin was also detected in the cerebrospinal fluid (CSF) of SHR and SHROB (13.8 and 27.2 pmol/L, respectively); however, the CSF/plasma ratio for leptin was 32 fold greater in SHR than in SHROB. To assess the putative action of leptin and leptin receptors on insulin-mediated glucose transport, muscles from SHR and SHROB were incubated in vitro with recombinant human leptin. Leptin directly suppressed insulin-mediated glucose transport by 50% in skeletal muscle from SHR but not in obese SHROB rats lacking all forms of the leptin receptor. These results suggest that the natural leptin receptor knockout in the SHROB represents a unique rat model to define the functional role(s) of leptin in central and peripheral energy metabolism. PMID- 9868175 TI - Probiotics reinforce mucosal degradation of antigens in rats: implications for therapeutic use of probiotics. AB - The effects of probiotics, administered with different diets, i.e., unhydrolyzed or hydrolyzed dietary antigens, on macromolecular degradation in the gut mucosa were studied. Rat pups were divided into five feeding groups at the age of 14 d. In addition to maternal milk, the milk group was gavaged daily with cows' milk and the hydrolysate group with extensively hydrolyzed whey formula, while controls received sterile saline. In addition to these diets, the milk-GG group and the hydrolysate-GG group were given probiotic bacteria, Lactobacillus GG ATCC 53103 (10(10) colony-forming units per day). At 21 d, the absorption of macromolecules, horseradish peroxidase and beta-lactoglobulin across patch-free jejunal segments was studied in Ussing chambers. The degree of macromolecular degradation was studied by means of HPLC gel filtration. The absorption rate of intact horseradish peroxidase differed among the feeding groups (P = 0.038). This was due to the high median (interquartile range) absorption of intact horseradish peroxidase (ng x h-1 x cm-2) in the milk group [255 (14-1332)] and supplementation with L. GG in the milk-GG group [35 (8-233)] restoring the status to the control level [22 (0-116)]. A parallel effect was seen in the hydrolysate group [100 (9-236)] vs. the hydrolysate-GG group [1 (0- 13)]. A gel filtration study confirmed that larger molecules were absorbed across the mucosa in the milk group compared to the other groups. The absorption of degraded horseradish peroxidase differed between the feeding groups (P = 0. 005). L. GG had a distinct effect when administered with unhydrolyzed, native protein vs. hydrolyzed protein: it increased absorption of degraded horseradish peroxidase in the milk GG group [7310 (4763-8228)] vs. the milk group [3726 (2423-5915)], while reducing it in the hydrolysate-GG group [2051 (1463-2815)] vs. the hydrolysate group [4573 (3759-9620)]. Our results showed that probiotics not only restore aberrant macromolecular transport, but they also have a specific effect on mucosal degradation depending on dietary antigen: adjuvant-like properties (unhydrolyzed antigen) and immunosuppressive-like properties (hydrolyzed antigen). The antigenicity of the diet therefore should be taken into consideration, when introducing novel probiotic functional foods for the management of gastrointestinal disorders. PMID- 9868176 TI - Antihyperglycemic actions of Eucalyptus globulus (Eucalyptus) are associated with pancreatic and extra-pancreatic effects in mice. AB - Eucalyptus globulus (eucalyptus) is used as a traditional treatment for diabetes. In this study, incorporation of eucalyptus in the diet (62.5 g/kg) and drinking water (2.5 g/L) reduced the hyperglycemia and associated weight loss of streptozotocin-treated mice. An aqueous extract of eucalyptus (AEE) (0.5 g/L) enhanced 2-deoxy-glucose transport by 50%, glucose oxidation by 60% and incorporation of glucose into glycogen by 90% in mouse abdominal muscle. In acute, 20 min incubations, 0.25-0.5 g AEE/L evoked a stepwise 70-160% enhancement of insulin secretion from the clonal pancreatic beta-cell line (BRIN-BD11). The stimulatory effect of 0.5 g/L AEE was unaltered by the presence of 400 micromol diazoxide/L and prior exposure to AEE did not alter subsequent insulin secretory response to L-alanine, thereby negating adetrimental effect on cell viability. The effect of AEE was not potentiated by glucose or demonstrable in cells exposed to a depolarizing concentration of KCl. Further study of the insulin-releasing effects of AEE revealed the activity to be heat stable, acetone insoluble, stable to acid, but abolished by exposure to alkali. Sequential extraction with solvents revealed activity in both methanol and water fractions, indicating the presence of more than one biologically active extract constituent. These data indicate that Eucalyptus globulus represents an effective antihyperglycemic dietary adjunct for the treatment of diabetes and a potential source for discovery of new orally active agent(s) for future therapy. PMID- 9868177 TI - Natural dietary polyphenolic compounds cause endothelium-dependent vasorelaxation in rat thoracic aorta. AB - This study investigated the possible active principles which support the endothelial nitric oxide-dependent relaxation produced by red wine and other plant polyphenolic compounds in thoracic aorta from male Wistar rats (12-14 wk old). Relaxation experiments were recorded isometrically on vessels precontracted with norepinephrine. Ten different chromatographic fractions (3-18 mg) isolated from red wine polyphenolic compounds (RWPC) and some available defined polyphenols (10-15 mg) were tested. Fractions enriched into either anthocyanins or oligomeric condensed tannins exhibited endothelium-dependent vasorelaxant activity (maximal relaxation in the range of 59-77%) comparable to the original RWPC. However, polymeric condensed tannins elicited a weaker vasorelaxant activity than the original RWPC (maximal relaxation ranged between 20-47%, P < 0.01). Moreover, the representative of either phenolic acid derivatives (benzoic acid, vanillic acid, gallic acid), hydroxycinnamic acid (p-coumaric acid, caffeic acid) or the flavanol [(+)-epicatechin] classes failed to induce this type of response. Among the anthocyanins, delphinidin (maximal relaxation being 89%), but not malvidin or cyanidin, showed endothelium-dependent vasorelaxation. These results show that anthocyanins and oligomeric-condensed tannins exhibited a pharmacological profile comparable to the original RWPC. These compounds may be involved in the reduction of cardiovascular mortality related to the presence of wine, fruits and vegetables in the diet. PMID- 9868179 TI - Maternal and fetal insulin-like growth factor system and embryonic survival during pregnancy in rats: interaction between dietary chromium and diabetes. AB - Chromium (Cr) depletion may exacerbate hyperglycemia and negative outcomes of pregnancy in the streptozotocin (STZ) diabetic pregnant rat model through the regulation of the insulin-like growth factor (IGF) system. To test this hypothesis, 40 female rats, all fed a low Cr diet (i.e., 70 microgram Cr/kg diet ) from 21 d of age, were randomly assigned one of four treatments, applied on Day 1 of pregnancy, in a 2 x 2 factorial design: 1) very low Cr diet (40 microgram Cr/kg diet) + citrate buffer injection, 2) very low Cr diet + STZ injection (30 mg STZ/kg body wt in citrate buffer), 3) adequate Cr diet (2 mg Cr [from added CrK(SO4)2]/kg diet) + citrate buffer injectionand 4) adequate Cr diet + STZ injection. Blood and tissues were collected on Day 20 of pregnancy. Chromium depletion increased (P < 0.05) urinary hydroxyproline excretion, 22-kDa IGF binding protein (IGFBP) concentration and litter size but decreased (P < 0. 05) placental wt, percentage of protein per fetus, and fetal IGF-I and -II concentrations. Chromium had no effect (P > 0.10) on maternal hormones, 32-kDa IGFBP, glucose, or placental and fetal hydroxyproline concentrations. Diabetes decreased (P < 0.05) maternal wt gain, embryonic survival, litter size, mean pup wt and maternal insulin concentrations, increased (P < 0.05) maternal blood glucose, IGF-I concentrations and maternal hydroxyproline excretion but did not affect fetal concentrations of hormones, IGFBP, glucose or hydroxyproline. Interaction between chromium and diabetes tended (P < 0.10) to affect maternal IGF-II concentrations, but had no effect on other maternal or fetal variables. In conclusion, maternal chromium depletion did not exacerbate hyperglycemia or pregnancy outcome in STZ-induced diabetic rats, but may negatively affect fetal protein content by decreasing fetal IGF-II concentrations. Diabetes may negatively affect fetal growth through its effect on maternal glucose, insulin and IGF-I. PMID- 9868178 TI - Green tea polyphenols block endotoxin-induced tumor necrosis factor-production and lethality in a murine model. AB - Green tea polyphenols are potent antioxidants. They have both anti-cancer and anti-inflammatory effects. However, their mechanisms of actions remain unclear. In inflammation, tumor necrosis factor-alpha(TNFalpha) plays a pivotal role. NF KB, an oxidative stress -sensitive nuclear transcription factor, controls the expression of many genes including the TNFalpha gene. We postulated that green tea polyphenols regulate TNFalpha gene expression by modulating NF-KB activation through their antioxidant properties. In the macrophage cell line, RAW264.7, ( )epigallocatechin gallate (EGCG), the major green tea polyphenol, decreased lipopolysaccharide (LPS)-induced TNFalpha production in a dose-dependent fashion (50% inhibition at 100 mmol/L). EGCG also inhibited LPS-induced TNFalpha mRNA expression and nuclear NF-KB-binding activity in RAW264.7 cells (30-40% inhibition at 100 mmol/L). Similarly, EGCG inhibited LPS-induced TNFalpha production in elicited mouse peritoneal macrophages. In male BALB/c mice, green tea polyphenols (given by oral gavage 2 h prior to an i.p. injection of 40 mg LPS/kg body wt) decreased LPS-induced TNFalpha production in serum in a dose responsive fashion. At a dose of 0.5 g green tea polyphenols/kg body wt, serum TNFalpha was reduced by 80% of control. Moreover, 0.5 g green tea polyphenols/kg body wt completely inhibited LPS-induced lethality in male BALB/c mice. We conclude that the anti-inflammatory mechanism of green tea polyphenols is mediated at least in part through down-regulation of TNFalpha gene expression by blocking NF-KB activation. These findings suggest that green tea polyphenols may be effective therapy for a variety of inflammatory processes. PMID- 9868180 TI - Liver alpha-tocopherol transfer protein and its mRNA are differentially altered by dietary vitamin E deficiency and protein insufficiency in rats. AB - To study how the expression of alpha-tocopherol transfer protein (alpha-TTP) and its mRNA are affected by protein and vitamin E status, Long-Evans male weanling rats were fed a vitamin E-deficient (DE), high vitamin E (HE, 5 g/kg diet of all rac-alpha-tocopheryl acetate) or control (C) diet for 12 wk in Experiment 1; and fed a low-protein (LP) or control (C) diet for 6 wk in Experiment 2. The high and deficient vitamin E status of HE and DE groups in Experiment 1 were confirmed by changes in plasma pyruvate kinase activity as well as the concentrations of alpha tocopherol in plasma and liver. As shown by the Northern and Western Blot Analysis, the expression of alpha-TTP in the liver of the DE group was significantly lower than, while that of the HE group was not different from, that of the controls. In contrast, the alpha-TTP mRNA levels did not differ among the C, DE and HE groups. alpha-Tocopherol in most peripheral tissues of rats fed the LP diet in Experiment 2 was significantly lower than that of the C. Both the alpha-TTP and its mRNA were significantly lower in the LP group than in the C. The results suggested that dietary vitamin E does not affect alpha-TTP gene expression except that the protein levels in the liver were lowered by vitamin E deficiency. On the other hand, protein inadequacy appeared to down-regulate the expression of the alpha-TTP gene. PMID- 9868181 TI - Supplement users differ from nonusers in demographic, lifestyle, dietary and health characteristics. AB - This study delineates demographic, lifestyle, dietary and health factors associated with the use of supplements at varying levels. Data are from a population-based cohort of 2,152 middle- to older-age adults living in Beaver Dam, Wisconsin. Information was collected by in-person interviews between 1988 1990. Associations were adjusted for gender and age. Use of supplements was more prevalent among women, persons with more than 12 years of education, those with relatively low body mass indices, persons with active lifestyles, and persons who never smoked as compared to current smokers (P 0.10) although uptake varied 3-fold over 24 h. Mitochondrial lysine uptake was greater (P < 0.05) for rats fed diets containing 60% casein than for rats fed diets containing 5% casein. Diurnal variation of LKR was detected (P < 0. 05) in rats fed diets containing 20 and 60% casein. Diurnal variation of SacD was detected (P < 0.05) in rats fed diets containing 60% casein. Increased casein consumption resulted in increased LKR and SacD activities (4- to 5-fold; P < 0.05). Diurnal variation of LOX was detected in rats fed diets containing 20 and 60% casein (P < 0.05). Increasing the casein concentration in the diet from 5 to 60% resulted in a 7-fold increase in LOX (P < 0.05). To make rate comparisons, LKR and SacD activities and LOX were predicted from a range of substrate concentrations (0.1 to 5.0 mmol/L). Overall, LKR and SacD were 6-107 times that of LOX, suggesting that, in liver, mitochondrial lysine uptake limits LOX. PMID- 9868192 TI - Infusion of soy and casein protein meals affects interorgan amino acid metabolism and urea kinetics differently in pigs. AB - For routine evaluation of the quality of dietary protein, amino acid scoring patterns were used. Evaluation of this pattern for soy and casein revealed that these proteins are of almost equal quality. However, in vivo studies showed a large difference. To study the biological effects of meals with casein and soy protein, the contributions of individual amino acids to net protein retention and amino acid kinetics in gut, liver and muscle in healthy pigs were investigated. Isonitrogenous enteral nutrition, infused at a rate of 10 mL. kg body wt-1. h-1 and consisting of maltodextrin (137 g/L) with added casein (53 g/L) or soy protein (68 g/L), was given to conscious, healthy female multicathetized pigs (20 22 kg, n = 12). A primed-constant infusion protocol with L-[ring-2,6 3H]phenylalanine, L-[3,4-3H]valine and [15N-15N]urea was used to measure amino acid and urea kinetics in gut, liver and muscle. Measurements were done postabsorptively and 2-6 h after initiation of the enteral nutrition. During the meal, appearance of amino acids into the portal vein and the uptake by the liver was lower with casein infusion. Muscle uptake did not differ. Gut protein synthesis tended to be lower with soy infusion (P = 0.1). Liver protein synthesis and degradation were higher with casein infusion (P < 0.05), while in muscle, soy infusion stimulated protein turnover (P < 0.05). In comparison to the postabsorptive condition, liver urea production was unchanged after casein infusion, while it was significantly increased after soy infusion. These results suggest that the quality of soy protein is inferior to that of casein protein. PMID- 9868193 TI - Absorption of nutrients is only slightly reduced by supplementing enteral formulas with viscous fiber in miniature pigs. AB - Viscous polysaccharides reduce intestinal absorption of glucose and diminish postprandial hyperglycemia. However, it is unknown whether viscous fiber also inhibits absorption of nutrients under conditions of enteric feeding. Therefore, we measured the absorption rates of nutrients in miniature pigs by perfusing a 150-cm length of jejunum with 8.37 kJ/min of the three following enteral diets: an isoosmotic oligomeric diet (1670 kJ/L), a hyperosmotic oligomeric diet and an isoosmotic polymeric diet (both 3350 kJ/L). The diets were supplemented with guar gum from 0 to 4.4 g/L. With the three guar-free diets, the mean absorption rate of energy was 5.2 +/- 0.32 kJ/min, corresponding to 62% of the energy infused. Absorption rates of carbohydrate, protein, fat and energy linearly declined as concentrations of guar or the logarithm of chyme viscosity increased. Due to modulations in viscosity, the inhibitory effects of guar were significantly different among the three diets. With the isoosmotic and hyperosmotic oligomeric and the polymeric diets, the addition of 1 g guar/L diminished the absorption of energy by 9.7, 6. 6 and 3.7%, respectively. The strong inhibitory effect on nutrient absorption with the isoosmotic oligomeric diet was caused by an increase in chyme viscosity due to water absorption. With the hyperosmotic oligomeric and the polymeric diets, the chyme viscosity and thus inhibitory effects on absorption were diminished by water secretion and the concomitant infusion of pancreatic enzymes. Results indicate that the addition of small amounts of guar gum to enteral diets of high energy density exerts only small effects on absorption of nutrients. PMID- 9868194 TI - Maternal blood, egg and larval thiamin levels correlate with larval survival in landlocked Atlantic salmon. AB - A link was previously established between the Cayuga syndrome, a condition causing 100% mortality in larval landlocked Atlantic salmon, Salmo salar, in several of New York's Finger Lakes, and a maternal diet of alewife, Alosa pseudoharengus, a non-native thiaminase-rich Clupeid fish. We evaluated salmon larvae viability relative to maternal thiamin status, and investigated the putative link of the Cayuga syndrome to an alewife diet in fish from the geographic regions outside the Finger Lakes/lower Great Lakes watersheds. We identified Cayuga syndrome in Atlantic salmon from Otsego Lake in the Susquehanna River watershed and from Green Pond in New York's Adirondack Mountains. In both systems alewife represent the major component of the diet for the salmon. Thiamin levels in the maternal blood of Otsego salmon with syndrome-negative progeny were three- to four-fold greater than those Otsego females whose progeny exhibited 100% mortality. Thiamin levels in eggs and larvae were directly related to thiamin levels in maternal blood in both syndrome-positive and syndrome-negative stocks. Thiamin bath treatments of syndrome-afflicted larvae eliminated mortality regardless of their lake stock of origin. Maternal blood levels of approximately 0.31 nmol thiamin pyrophosphate/g or 0.44 nmol total thiamin/g appear necessary to achieve egg threshold levels of approximately 0.8 and 1.1 nmol/g unphosphorylated and total thiamin, respectively; these egg thiamin levels should prevent significant syndrome-related mortality in landlocked Atlantic salmon larvae. These results confirm the role of thiamin in the etiology of the Cayuga syndrome and support the dietary link of this naturally occurring thiamin deficiency to the thiaminase-rich alewife. PMID- 9868195 TI - In well-fed young rats, lactose-induced chronic diarrhea reduces the apparent absorption of vitamins A and E and affects preferentially vitamin E status. AB - To establish the effect of lactose-induced diarrhea on the apparent absorption and status of vitamins-A and E in well-fed young rats, we fed Sprague Dawley rats a balanced diet or a lactose diet (350 g/kg). A group of rats fed the control diet equal to the level measured in the lactose-fed rats (pair-fed) was also included. The experiment lasted 23 d and feces were collected on days 4-6, 10-12, 14-16 and 20-22. Samples of serum and tissues were taken on days 10 and 23. Lactose caused a significant reduction in food intake, had no effect on body weight and produced a diarrhea that persisted during the whole experiment. The severity of diarrhea decreased with time, indicating that the rats partially adapted to lactose feeding. At the onset of diarrhea, the apparent absorption of vitamins A and E in the rats with diarrhea was significantly lower than in the control or pair-fed rats, but the rats with diarrhea recovered gradually, and in the case of vitamin E was normalized by day 15. At day 10 the rats with lactose induced diarrhea had serum and liver concentrations of vitamins A and E that did not differ from the control or pair-fed rats. However, at day 23 the lactose-fed rats with diarrhea had significantly lower serum and liver concentrations of vitamin E than the control or pair-fed rats. Measured at that time, diarrhea had no effect on liver vitamin A, but lower serum concentrations of this vitamin were detected in both the lactose-fed rats and in the pair-fed rats. In general, in well-nourished rats, the chronic diarrhea associated with excessive dietary lactose reduced the apparent absorption of vitamin A and E and particularly compromised the nutritional status of vitamin E. PMID- 9868196 TI - Long-chain polyunsaturated fatty acid levels in formulae influence deposition of docosahexaenoic acid and arachidonic acid in brain and red blood cells of artificially reared neonatal rats. AB - We studied the effects of dietary long-chain polyunsaturated fatty acids (PUFA) on the fatty acid composition of the brain and red blood cells in gastrostomized rat pups reared artificially from postnatal Days 5-18. These pups were fed rat milk substitutes in which the fat comprised 10% linoleic acid and 1% alpha linolenic acid and, using a 3 x 3 factorial design, one of three levels of both arachidonic acid (AA) and docosahexaenoic acid (DHA) supplied as single cell microbial oils (0.0, 0.4 and 2.4% fatty acids). A tenth group was reared by nursing dams. The fatty acid composition of the phosphatidylethanolamine (PE) and phosphatidylserine/phosphatidylinositol (PS/PI) phospholipids in the brain and red blood cells on Day 18 reflected the dietary composition in that pups receiving long-chain supplementation of each had higher levels of the supplemented PUFA, but lower levels of the other, relative to unsupplemented groups. In contrast to these results, there were few changes in the brain in phosphatidylcholine (PC) phospholipids whereas, in the red blood cells, changes in PC were similar to those in PE and PS/PI. Regression analyses showed that DHA levels in the brain correlated more closely with those of the red blood cells than did AA levels. The results of this study indicate that, although supplementation of formula with AA or DHA during the period of rapid brain development in rats increases deposition of the long-chain PUFA in the developing tissues, each also affects the levels of the other. PMID- 9868197 TI - Boron stimulates embryonic trout growth. AB - Boron is present in our soil, water and air. Cyanobacteria require it for nitrogen fixation, and vascular plants require it for the formation of cell walls and membranes. I report here how boron affects the growth of embryonic rainbow trout (Oncorhynchus mykiss). Fertilized ovum from the Mt. Whitney rainbow trout strain were incubated at (12.5 degreesC) in Type 1 ASTM ultrapure grade water supplemented with boric acid (99.5% purity) during the 1995 and 1997 spawning seasons. Boron concentrations of the incubation solutions were determined by direct measurement using the curcumin procedure or inductively coupled plasma mass spectrometry. In the 1995 study boron ranged from 1 to 936 micromol/L. Ca, Na and Mg salts were included in the incubation solutions to approximate concentrations in natural water. In the 1997 study fertilized eggs were incubated in ultrapure water supplemented with boric acid alone over a range from 2.2 to 90.6 micromol/L. The 1995 study used 144 embryos per B concentration and the 1997 study used 96 embryos per B concentration. Growth and teratogenicity were evaluated at the eye, hatch and 2-wk posthatch developmental stages. Boron stimulated growth in a dose-dependent manner in both studies (P < 0.001), and exposure was associated with an increase in B body concentration (P < 0.05). No teratogenic or microbicidal effects were apparent. These results are consistent with those expected of an element essential for vertebrate development. J. Nutr. 2488-2493, 128: 1998 PMID- 9868198 TI - Dietary biotin requirement for maximum growth of juvenile grass shrimp, Penaeus monodon. AB - A feeding trial was conducted to estimate the minimal dietary biotin requirement for juvenile grass shrimp, Penaeus monodon. Purified diets with eight levels (0, 0.2, 0.5, 1.0, 3.0, 6.0, 10.0 and 20.0 mg/kg) of supplemental biotin were fed to P. monodon (mean weight 0. 26 +/- 0.01 g) for 8 wk. Each diet was fed to three replicate groups of shrimp. Shrimp fed diets supplemented with biotin (0.2-20.0 mg/kg) had significantly (P < 0.05) higher weight gain, feed efficiency and protein efficiency ratio than those fed the unsupplemented control diet. Weight gain was high in shrimp fed 3. 0-10.0 mg biotin/kg diet and lowest in shrimp fed 50% of energy) diets increase stamina and maximize energy production, and high protein (>30% of energy) diets prevent training-induced anemia. Nutrient requirements differ, however, for sprint racing dogs, such as greyhounds. Greyhounds run faster when fed moderately increased dietary fat but run more slowly when dietary protein is increased. Sled dogs have similar energy requirements to other breeds at rest in a thermoneutral environment ( approximately 550W0.75 kJ/d where W is body weight in kg) but may require as much as 4200W0.75 kJ/d during a race. The energy requirement of greyhounds in training, however, is only approximately 600W0.75 kJ/d. There is little information, however, concerning the vitamin, mineral or other nutrient requirements of athletic dogs; most sled dogs and greyhounds are fed "homemade" recipes. These recipes usually include raw meat and represent a health risk. More studies are required to improve the health and performance of working and racing dogs. PMID- 9868243 TI - Changes in plasma and muscle creatine concentration after increases in supplementary dietary creatine in dogs. PMID- 9868244 TI - Biological and nutritional consequences of work at high altitude in search and rescue dogs: the scientific expedition Chiens des Cimes-Licancabur 1996. PMID- 9868245 TI - Developments in equine nutrition: comparing the beginning and end of this century. AB - In the first part of this century, there was little advancement in horse nutrition and little research was undertaken. However, in the last few years, there has been a great increase in interest in this whole area. This review surveys some of the more recent developments and how they have influenced feeding practices and also compares these with those feeding practices found at the start of this century. The review concentrates on the nutrition of the adult horse in work, exploring in particular what they are fed and how the nutrient value of these feeds is evaluated. PMID- 9868246 TI - Exercise affects digestibility and rate of passage of all-forage and mixed diets in thoroughbred horses. PMID- 9868248 TI - Periodontal disease and diet in domestic pets. AB - Periodontal disease is the most common oral condition seen in domestic pets. In addition to the discomfort caused in the affected animal, there is strong circumstantial evidence to show that a focus of infection in the mouth may cause disease of distant organs. Consequently, prevention of periodontal disease is of paramount importance for the general health and well-being of pets. The presence of plaque on the tooth surfaces is the primary cause of periodontal disease. However, the mechanisms by which disease develops are by no means fully understood. Dietary texture does have an effect on the accumulation of dental deposits and consequently on disease development and progression. Daily toothbrushing remains the single most effective means of removing plaque, thus preventing periodontal disease. On the basis of current knowledge, the best way to maintain healthy periodontal tissues in our pets is frequent toothbrushing. The use of a dental hygiene chew and/or a diet designed to reduce dental deposits can be useful adjunctive measures and should be recommended. PMID- 9868247 TI - Dietary carbohydrates and fat influence milk composition and fatty acid profile of Mare's milk. PMID- 9868249 TI - Halitosis in dogs and the effect of periodontal therapy. PMID- 9868250 TI - Diet and large intestinal disease in dogs and cats. AB - Large intestinal disease, and more especially colitis, is a commonly seen problem in small animal practice. Although colitis is most frequently diagnosed in dogs, it is becoming increasing common in cats. The etiology of colitis is not known, but there is general agreement that an immune-mediated response to luminal antigen is involved. In particular, parasites, bacteria and dietary factors may be involved. In approximately 10% of dogs presented with typical signs of colitis, no pathologic lesion will be found on investigation. These dogs have a functional diarrhea associated with some stress factor and are thought to have irritable bowel syndrome (IBS). This condition is most frequently observed in working dogs, although highly nervous and excitable dogs may also exhibit similar clinical signs. Until the underlying etiology of colitis is determined, treatment regimens will remain symptomatic. Recent studies have placed considerable importance on the value of diet in the prevention, immediate and long-term therapy of colitis in dogs and cats. In particular the value of "novel" protein diets, fermentable fiber and polyunsaturated fatty acids is receiving the most attention. It is now possible to maintain patients in long-term remission and to modify the severity and chronicity of colitis by using diet alone. This paper will review the subject of dietary management of colitis and IBS and present results from the author's clinical research program. PMID- 9868251 TI - Intravenous glutamine or limited enteral feedings in piglets: amelioration of small intestinal disuse atrophy. PMID- 9868252 TI - Atrophied small intestinal responses of piglets to oral feedings of milk. PMID- 9868253 TI - The use of probiotics in the diet of dogs. PMID- 9868254 TI - Nutritional support for dogs and cats with hepatobiliary disease. AB - Nutritional intake in the patient with hepatobiliary disease provides the cornerstone of balanced medical care. Optimal recommendations require consideration of general nutritional principles, special species requirements and contemporary needs uniquely related to the patient's medical problem. Although general recommendations follow well-established guidelines developed to meet metabolic requirements for normal health, there is little information regarding altered requirements in animals that are ill. Consequently, recommendations for animals have been derived empirically from studies completed in humans, most work having been done in patients with end stage cirrhosis or liver failure complicated by hepatic encephalopathy. This is problematic because most veterinary patients with liver disease are not in hepatic failure and do not suffer from hepatic encephalopathy. Iatrogenic malnutrition can develop in patients when protein-restricted diets are inappropriately recommended. Insufficient energy intake and negative nitrogen balance can complicate a patient's condition, impairing tissue regeneration and recovery from disease. This paper reviews strategies that can be used to individualize nutritional management in small companion animals with hepatobiliary disease. Consideration is given to both the known and controversial issues regarding energy requirements, dietary energy distribution, vitamin and micronutrient supplementation, the special requirements of the cat with hepatic lipidosis, as well as strategies effective for palliation of hepatic encephalopathy. PMID- 9868256 TI - Tumor necrosis factor-alpha levels in adipose tissue of lean and obese cats. PMID- 9868255 TI - Characterization of serum lipoprotein profiles of healthy, adult cats and idiopathic feline hepatic lipidosis patients. PMID- 9868257 TI - The effect of diet on lower urinary tract diseases in cats. AB - Because dietary ingredients and feeding patterns influence the volume, pH and solute concentration of urine, diet can contribute to the etiology, management or prevention of recurrence of some causes of lower urinary tract disease. Most research assessing the effect of diet has focused on the latter two aspects, primarily because of interest in struvite urolithiasis. Manipulation of urine pH through dietary means has proven an effective tool for the management and prevention of struvite urolithiasis; acidification of urine, however, may be a risk factor for calcium oxalate urolithiasis, which now appears to occur with approximately equal frequency in cats. Prediction of urine pH from dietary analysis would thus be a valuable tool, but considerable further research is required before this can be achieved with commercial canned foods. With the growing importance of urolith types other than struvite, alternatives to the measurement of urine pH are required to assess critically the likely beneficial (or detrimental) effects of manipulation of nutrient profile. Measurement of urinary saturation may permit the development and fine tuning of nutrient profiles aimed at controlling lower urinary tract diseases in cats that are associated with a range of different mineral types. The majority of cats with signs of lower urinary tract disease do not, however, have urolithiasis; indeed, no specific cause can be established in most of these cats. Recent observations suggest that recurrence rates of signs in cats classified as having idiopathic lower urinary tract disease may be more than halved if affected animals are maintained on high, rather than low moisture content diets. J. Nutr. 2753S-2757S, 1998 PMID- 9868258 TI - Accuracy of quantitative collection of urine in carnivores. PMID- 9868259 TI - A system to monitor urinary tract health in dogs. PMID- 9868260 TI - Urinary relative supersaturations of calcium oxalate and struvite in cats are influenced by diet. PMID- 9868261 TI - Is there a role for dietary polyunsaturated fatty acid supplementation in canine renal disease? AB - Dogs with spontaneous renal diseases frequently develop progressive uremia. After partial nephrectomy, a similar pattern of progressively declining renal function develops. This pattern may be attributed in part to the development of glomerular hypertension in remnant canine nephrons. Changes in the composition of dietary polyunsaturated fatty acids (PUFA) modify glomerular hemodynamics in normal rats and affect the chronic course of renal disease in partially nephrectomized rats. Thus, dietary PUFA supplementation might alter progressive canine nephropathies. However, the response of dogs with renal insufficiency to dietary manipulations frequently differs substantially from that of laboratory rodents, and the effects of dietary PUFA composition have been poorly characterized in dogs with chronic renal disease. Here we address the hypothesis that dietary PUFA supplementation may delay the progression of chronic renal insufficiency in dogs. In particular, dogs ingesting diets supplemented with (n-6) PUFA exhibited severe glomerular hypertension associated with rapidly progressive renal failure. In contrast, dietary supplementation with (n-3) PUFA prevented deterioration of the glomerular filtration rate and preserved renal structure. The results of these model studies demonstrate that dietary PUFA supplementation may alter renal hemodynamics and the long-term course of renal injury in dogs. Clinical trials to address the potential benefits of dietary (n-3) PUFA supplementation in a variety of spontaneous renal diseases seem warranted. PMID- 9868262 TI - Antioxidant status in dogs with idiopathic dilated cardiomyopathy. PMID- 9868263 TI - Advances in dietary management of obesity in dogs and cats. AB - Recent evidence in humans has reemphasized the importance of specific lifestyle behaviors such as activity level on energy requirements. A recent survey of adult pet dogs has shown a clear association between the level of activity and energy requirement, and suggests that current feeding recommendations may overestimate the energy requirements of adult dogs. Although a reduction in feeding guides may help to reduce the risk of overfeeding and subsequent development of obesity in adult dogs, there is considerable individual variation in energy requirements, which emphasizes the importance of tailoring feeding practices to the individual. Diet clearly has a critical role to play in both the prevention and treatment of obesity. We have evaluated the effect of different dietary regimens on the treatment of obesity in companion animals. In cats, increased energy restriction results in more rapid weight loss. However, this is associated with less favorable changes in body composition. In dogs, we have evaluated the potential benefit of insoluble and soluble dietary fiber on satiety in dogs that have been restricted to an energy intake appropriate for weight reduction. Results of a series of studies have failed to show any benefit of either fiber type on satiety in energy-restricted dogs. PMID- 9868264 TI - Diet and feline obesity. PMID- 9868265 TI - A comparison of the feeding behavior and the human-animal relationship in owners of normal and obese dogs. PMID- 9868266 TI - Diet and skin disease in dogs and cats. AB - Dietary factors have a major role in the maintenance of healthy coat and skin, and are significant in the etiology and therapy of certain skin diseases. Nutritional deficiencies are now uncommon as a result of the widespread feeding of complete and balanced pet foods. Deficiencies of (n-6) polyunsaturated fatty acids, zinc and vitamins, however, do arise in certain animal- or product-related instances. Supraphysiologic doses of vitamin A have been used in the management of vitamin A-responsive dermatosis in Cocker spaniels; other keratinization defects and seborrheic conditions may respond to retinoid therapy. Much interest has been paid to the therapeutic value of polyunsaturated fatty acid supplements in the management of dermatologic conditions associated with hypersensitivity reactions or keratinization defects. These studies have generally yielded disappointing results, which may reflect shortcomings in the design of some trials. Nevertheless, a placebo-controlled, double-blind, cross-over study has demonstrated a clear benefit of high dose (n-3) fatty acids in the management of pruritic skin disease. There is also preliminary experimental evidence that specific dietary (n-6):(n-3) fatty ratios are useful in the dietary management of inflammatory diseases. Although results of controlled clinical trials are awaited, the argument exists that it is the absolute amount of (n-3) fatty acid intake rather than ratio that is responsible for potential health benefits. PMID- 9868267 TI - Prevalence and causes of food sensitivity in cats with chronic pruritus, vomiting or diarrhea. PMID- 9868268 TI - Possible food allergy in a colony of cats. PMID- 9868269 TI - Comparison of various methods of fiber analysis in pet foods. PMID- 9868270 TI - Fiber affects digestibility of organic matter and energy in pet foods. PMID- 9868271 TI - Apparent ileal nitrogen and amino acid digestibility of a moist cat food. PMID- 9868272 TI - Selected fructooligosaccharide composition of pet-food ingredients. PMID- 9868273 TI - An improved method for the estimation of energy in pet foods. PMID- 9868274 TI - A comparison of the bioavailability of three dietary zinc sources using four different physiologic parameters in dogs. PMID- 9868275 TI - Raw and rendered animal by-products as ingredients in dog diets. PMID- 9868276 TI - Bone and cytokines: beyond IL-1, IL-6 and TNF-alpha. PMID- 9868277 TI - The microscopic structure of bone in normal children and patients with osteogenesis imperfecta: a survey using backscattered electron imaging. AB - The microstructure of iliac crest biopsies from normal children or from those afflicted with osteogenesis imperfecta (OI) has not previously been studied to determine the tissue histology in the context of the degree of mineralization. The material in this study comprised 112 iliac crest biopsies from children aged 1.9-22.9 years. Fifty-eight were reference biopsies taken from children with no bone disease and the remainder were biopsies from children diagnosed as having OI (23 were Type I, 8 Type III, 18 Type IV, and 5 Type V). The specimens, which had been embedded in polymethylmethacrylate (PMMA), were micromilled and carbon coated to permit backscattered electron imaging. Reference biopsies from very young children often contained densely mineralized cartilage, and evidence of rapid cortical drift. Circumferential lamellae became a prominent feature after the toddler stage, and active remodeling and slower cortical drift continued through childhood. The biopsies from older teenagers and young adults were indistinguishable. Occasional mineralized osteocyte lacunae were detected in even the youngest children. Bone from children with OI Type I often appeared normal in microstructure and amount, but in some there was a dearth of bone and an abundance of osteocytes. Compared with age-matched controls, cortical and trabecular bone from children with OI Types III and IV were markedly sparse and very cellular, and primary osteonal systems continued to be formed later than expected. A distinguishing feature of the bone from OI Type V patients was the failure of patches of bone to mineralize, especially adjoining a reversal line. Packets of bone tissue exhibiting either considerably higher than normal or deficient mineralization would contribute to the characteristic trait of mechanical weakness. PMID- 9868279 TI - Evaluation of short-term precision for tibial ultrasonometry. AB - Tibial quantitative ultrasonometry is a relatively novel technique in the field of bone sonometry, an emerging alternative to bone densitometry. The implementation of this technique in a pediatric population could prove valuable from a clinical as well as a research viewpoint. In clinical practice it is necessary to know the precision of this technique and the possible influence on measurements before implementation. This study presents the precision in a Caucasian pediatric population and the influence of measurement site, dexterity, brand of coupling gel, and temperature of coupling gel. To assess intra- and interobserver variance duplicate measurements, with repositioning, ultrasonometry was performed in 10 children over a short period of time. The observers were blinded for the results of the other observer and after each measurement the skin markings were removed. Intraobserver variance for operator one (MHL) and for observer two (SFGR) was CV 0.43%. The interobserver variance was CV 0.61%. Left midtibial and right midtibial speed of sound (SOS) measurements showed no significant differences. There were, however, significant differences in both boys and girls between right proximal versus right midtibial, right midtibial versus right distal, and right proximal versus right distal (for all P < 0.001). One-way analysis of variance (ANOVA) showed that neither the use of different coupling gels nor an increase in gel temperature had a significant influence on measurements. The results of our study show that tibial quantitative ultrasonography (QUS) is a highly reproducible technique in a Caucasian pediatric population. PMID- 9868278 TI - Ultrasound velocity of trabecular cubes reflects mainly bone density and elasticity. AB - Studies have indicated that quantitative ultrasound (QUS) variables may be influenced by the mechanical properties of bone which in turn are determined by bone's material and structural properties. However, from these studies it is unclear what role density, elasticity, and structure play in determining velocity. Eighteen defatted, 12-mm cubic trabecular bone specimens were cut from cadaveric specimens. Amplitude-dependent speed of sound (SOS) using a single point QUS system was assessed in three orthogonal axes. Magnetic resonance images were obtained, from which measures of apparent trabeuclar structure were derived. The specimens were nondestructively tested in compression along three orthogonal axes defined by the sides of the cubes. The elastic modulus (in the three directions) and the strength (in one direction) were determined. Trabecular BMD was measured by quantitative computed tomography. SOS varied significantly with direction of measurement, with the highest value in the axial direction (axial:1715 m/s, sagittal: 1662 m/second, and coronal: 1676 m/s). SOS of each of the three axes was generally associated with the various mechanical (r = 0.30 0.87), density (r = 0.81-0.93), and bone structural variables (0.3-0.8). However, after adjusting the SOS correlations by density, only the correlation with elasticity remained significant in the coronal direction. BMD alone explained 88 93% of variance in SOS whereas in the multivariate model, BMD plus elasticity and/or anisotropic variables explained 96-98% of the variance in SOS. Variability of SOS is explained mostly by density and to a small extent by elasticity or anisotropy. Since only 2-6% of the variance of the QUS measurement is not explained by density and elasticity, one could conclude that the remaining variance reflects other properties of bone or perhaps simply measurement error. Evidence that these other properties may be structure related is only found in the anisotropy of QUS parameter. PMID- 9868280 TI - Ultrasound velocity through the phalanges in normal and osteoporotic patients. AB - Quantitative ultrasound (US) measurements have been shown to be a new technique assessing bone status. This study aimed to assess a new US instrument, the DBM Sonic 1200(R) (IGEA) which permits the measurement of the speed of sound in the proximal phalanges (SOSp) of the hand. The results obtained were compared with DXA (SOPHOS) and US measurements at the calcaneus (Achilles(R) LUNAR). The in vivo precision expressed by coefficient of variation was 0.91%. Ultrasound measurements of phalanges were significantly correlated with BMD in the entire group of 90 subjects: osteoporotic patients (n = 47) and controls (n = 43) (r = 0.44, femoral neck and 0.45, lumbar spine, P < 0.01). A significant correlation was also found in the control group (r = 0.33, lumbar spine and 0.38, femoral neck, P < 0.05) but not in the osteoporotic group (r = 0.3, lumbar spine and 0.17, femoral neck, P > 0.05). Mean values for 31 postmenopausal, osteoporotic women and age-matched controls showed a significant decrease in US measurements at the phalanges (P < 0.05) and the calcaneus (P < 0.01) as well as bone mineral density (BMD) at the spine and femoral neck (P < 0.01) in the osteoporotic group. A decision threshold for a sensitivity of 80% for osteoporotic fractures resulted in a specificity value of only 37% for SOSp, between 53 to 65% for calcaneus US measurements and 45 to 56% for BMD. The Z score, the odds ratio, the ROC curves, and areas under the curves plotted for the subgroup of 31 fractures and their healthy controls showed poorer values for SOSp than BMD and calcaneus US measurements. In conclusion, US measurements of phalanges seem to be less efficient than calcaneus US and BMD measurements to distinguish osteoporotic from healthy women. Other studies and also prospective studies are required to assess the interest in fracture risk assessment. PMID- 9868281 TI - Measurement of forearm bone in children by peripheral computed tomography. AB - By means of peripheral computed tomography (pQCT), adult cortical bone density and volume was shown to be under a fixed rectilinear relationship regardless of age, sex, and presence or absence of osteoporosis. In children, however, the density-volume regression line followed a clearly different slope from that for adults (P < 0. 001), indicating a difference in property and composition of the cortical bone between growing bone of children and grown-up bone of adults. Although both relative cortical volume and density increased with age in both boys and girls, no significant increase of trabecular bone was noted in either during the growth period. When the same technique was applied to the bone of rats known to continue growing with indefinite modeling without remodeling, the regression line between cortical bone density and volume was different from that of adult humans and similar to that of growing human children. Growing bone in a constant process of modeling and mineralization thus seems to have cortical bone possibly with less complete mineralization somewhat different from those of grown up bone only undergoing remodeling. Selective cortical bone measurement by pQCT appears to be useful in characterizing the unique properties of the cortex of the growing bone. PMID- 9868282 TI - Age-related changes in body composition, hydroxyproline, and creatinine excretion in normal women. AB - We have made a cross-sectional study of relationships among age, whole body bone mineral content (WBBMC), and non-bone lean body mass (NBLBM) measured by dual energy X-ray absorptiometry (DXA), and daily excretion of hydroxyproline (OHP) and creatinine (Cr) in a group of normal women. WBBMC fell with age from the 6th decade, whereas NBLBM was almost constant. Creatinine excretion fell with age from the 5th decade until the 9th, to a much greater degree than NBLBM, reaching a nadir in the 8th decade. Daily excretion of hydroxyproline showed a peak in the 6th decade and fell moderately thereafter. The greater fall of creatinine compared with hydroxyproline resulted in rising OHP/Cr ratios with advancing age, in contrast to the pattern of hydroxyproline excretion. The use of creatinine as a correction for urine dilution or for lean body mass (LBM) in assays for markers of bone turnover must therefore be viewed with caution. PMID- 9868283 TI - Efficient gene transfer into normal human skeletal cells using recombinant adenovirus and conjugated adenovirus-DNA complexes. AB - In order to assess efficient DNA gene transfer into human primary cell cultures derived from the skeleton we tested two viral-based procedures. First, replication-deficient recombinant adenoviruses (ADV) were used to infect post confluent human marrow stromal fibroblasts (HMSF) and human trabecular bone (HTB) cells. Both cell types were readily infected by modified adenoviral vectors carrying a reporter gene making this virus an attractive candidate to facilitate DNA gene transfer. In a second approach we coincubated DNA with ADV that had polylysine (PLL) covalently attached. With this ADV/PLL/DNA complex, very efficient gene transfer into multilayered HMSF and HTB cell cultures was observed, and DNA coincubated with unmodified ADV failed to be effectively transferred. These data imply that the covalently bound PLL more effectively binds exogenous DNA, resulting in a highly efficient internalization event in both cell types. Thus, this latter method has many advantages over conventional ADV gene transfer procedures. It is simple, rapid, and it does not require engineering of DNA into the viral genome, thereby allowing transfer of large fragments of DNA. PMID- 9868284 TI - TGFbeta1 regulates 25-hydroxyvitamin D3 1alpha- and 24-hydroxylase activity in cultured growth plate chondrocytes in a maturation-dependent manner. AB - Chondrocytes metabolize 25-(OH)D3 to the two active dihydroxylated forms of the secosteroid, 1,25-(OH)2D3 and 24,25-(OH)2D3. The aim of the present study was to examine the activity of the enzymes responsible for this metabolism, 1alpha hydroxylase and 24R-hydroxylase, and their regulation by TGFbeta1. Basal 1alpha- and 24R-hydroxylase activities were measured in homogenates of confluent, fourth passage rat costochondral resting zone and growth zone chondrocytes and mouse cortico-tubular cells (MCT) were used as a positive control. The cells were harvested and homogenized in buffer optimized to maintain the activity and stability of the hydroxylases. Homogenates were incubated for 90 minutes and 1alpha- and 24R-hydroxylase activities determined by measuring the conversion of [3H]-25-(OH)D3 to [3H]-1,25-(OH)2D3 and [3H]-24,25-(OH)2D3 using an HPLC with an inline radioisotope detector. Resting zone cells were also treated with various concentrations of recombinant human TGFbeta1 for 24 hours, and enzyme activity in total cell homogenates as well as 24-hydroxylase mRNA levels were determined. In addition, [3H]-1,25-(OH)2D3 and [3H]-24,25-(OH)2D3 released into the conditioned media by resting zone chondrocyte cultures in response to TGFbeta1 were measured. In culture, all three cell types were found to contain 1alpha- and 24R hydroxylase activities. Basal 1alpha-hydroxylase specific activity was significantly higher than 24R-hydroxylase specific activity in all cells. RT-PCR confirmed that resting zone and growth zone cells expressed mRNA for 24R hydroxylase. Treatment of resting zone cells with TGFbeta1 increased 24R hydroxylase mRNA levels in a dose-dependent manner. TGFbeta1 also increased 24R hydroxylase activity 2- to 5-fold and decreased 1alpha-hydroxylase activity by 20 30%. Similar changes were observed with MCT cells, but not growth zone cells. Production of [3H]-24,25-(OH)2D3 by resting zone cells increased with TGFbeta1 treatment, while [3H]-1,25-(OH)2D3 production decreased. The effect was time- and dose-dependent, correlating with hydroxylase activity and 24-hydroxylase gene expression. These results demonstrate that growth plate chondrocytes contain the necessary enzymes to produce 1, 25-(OH)2D3 and 24,25-(OH)2D3 from 25-(OH)D3. In addition, the activity of these enzymes in resting zone cells, but not growth zone cells, is regulated by TGFbeta1 by increasing gene transcription, indicating that cell maturation-dependent autocrine/paracrine pathways exist for regulating vitamin D metabolite production. PMID- 9868285 TI - Monocyte chemotactic protein-1 gene expression and translation in formed granulomatous calcified tissue in vivo. AB - Monocyte chemotactic protein-1 (MCP-1) and related molecules constitute the C-C class of the beta chemokine supergene family with inflammatory properties. However, the exact role, function, and implication in inflammatory diseases remain to be determined. Here we report that subcutaneous injections (0.2 ml) of a saturated water solution (1:40) of potassium permanganate crystals induces the generation of granuloma tissue at the site of injection in the rat, and reaches its peak of formation after 1 week. The size and weight of the granulomas were increased by i.p. lipopolysaccharide (LPS) (6 microgram/200 microliter) and inhibited by intraperitoneal (i.p.) dexamethasone (Dxs) 300 microgram/200 microliter) treatments in rats, injected 18 hours before sacrifice. Moreover, steady-state levels of MCP-1 mRNA in the granuloma tissue (control), were strongly generated. Rats treated i.p. with LPS produced an increase of MCP-1 mRNA in the granuloma tissue compared with controls (i.p. PBS-treated) whereas in animals treated with Dxs, there was a decrease in (P < 0.05) in formation of mRNA protein. When the granuloma tissues were homogenized the generation of MCP-1 was found in the supernatants. The level of MCP-1 was higher (P < 0.05) in the LPS treated animals and lower (P < 0.05) in the Dxs group compared with the controls (treated with PBS). Similar results were obtained in the serum and in minced granuloma tissue where samples were further incubated in vitro with LPS (100 ng/ml) overnight. A Strong increase (P < 0.01) in MCP-1 in all samples was detected, but not in the minced granuloma tissue from Dxs-treated animals. Our data demonstrate that calcified tissue from chronic inflammation induced by KMnO4 generates MCP-1 gene expression and translation, an effect increased by LPS and decreased by Dxs. PMID- 9868286 TI - Expression of BMP-2 by rat bone marrow stromal cells in culture. AB - To investigate the role of bone morphogenetic protein (BMP-2) in ossifying rat bone marrow stromal cell cultures, we determined the population of fibroblast like stromal cells that expressed BMP-2 immunocytochemically (anti-rhBMP-2 monoclonal antibody), and compared that to alkaline phosphatase (AP) and collagen synthesis formed in culture over a 4-week period in control and dexamethasone supplemented mineralizing media. In control media, the percentage of BMP-2 positive stromal cells (BMP-2(+)) increased from 12 to 25% within the first 4 days of culture. In mineralizing media, the level of BMP-2(+) cells was significantly increased (43-44%). The intensity of immunostaining gradually increased with time. The levels of AP were undetectable at 1 week in both control and mineralizing media, but increased gradually over the next 2 weeks and peaked at 3 weeks. ALP levels were significantly greater in cultures grown in mineralizing medium (P < 0.05 at 3 weeks, P < 0.01 at 4 weeks). Collagen synthesis peaked and was significantly greater at 3 weeks (P < 0.05) in cultures grown in mineralizing medium. The levels of AP and collagen synthesis most closely reflected the changes in the percentage of BMP-2(+) cells from 7 to 28 days. Though these changes may reflect a primary action of BMP-2 on marrow osteoprogenitor-like stromal cells, they do not exclude a mechanism that involves the induction of other members of the BMP family known to stimulate AP and collagen synthesis. We conclude that BMP-2 expression in cultures of fibroblast like marrow stromal cells is enhanced when those cells are induced to become osteoblasts by exposure to dexamethasone. PMID- 9868287 TI - Basic fibroblast growth factor in the presence of dexamethasone stimulates colony formation, expansion, and osteoblastic differentiation by rat bone marrow stromal cells. AB - Basic fibroblast growth factor (bFGF) is known to stimulate endosteal bone formation in vivo by a mechanism possibly mediated via osteoblast precursor cells present in the bone marrow. In high density cultures of primary bone marrow cells, and in the presence of glucocorticoids, bFGF stimulates the formation of a bone-like matrix; however, due to the dense nature of these cultures, the exact mechanism of action is unclear. In an adaptation of the fibroblastic colony formation unit assay, in which the bone marrow cells are grown in the presence of dexamethasone, beta-glycerophosphate, and ascorbate, mineralized colonies are formed which stem from single mesenchymal precursor cells and grow in isolation from each other. Using this system we have been able to investigate the mechanism by which bFGF stimulates the formation of bone like tissue in vitro. We have shown that bFGF increases the formation of a calcified collagenous matrix in vitro by (1) increasing the total number of fibroblastic colonies formed, (2) increasing the proportion of differentiated colonies that synthesize collagen and calcify, and (3) stimulating the proliferation and collagen accumulation of the individual colonies. A maximal increase in total and differentiated colony numbers was seen after only 5 days exposure to bFGF, however, continued exposure to bFGF continued to increase the size and collagen content of the individual colonies. Bearing in mind the endosteal location of newly formed bone seen after treatment with bFGF, these processes may well play an active role in this effect. PMID- 9868288 TI - Changes in the concentration of insulin-like growth factor I and transforming growth factor beta1 in rat femoral bone during growth. AB - Our knowledge of the concentration of growth factors in growing bone is limited. In the present study, we examined the developmental changes in the concentrations of insulin-like growth factor I (IGF-I) and transforming growth factor beta (TGF beta) in the rat femur between weanling and maturity. We show that during the rapid growth phase there is a continuous rise in bone matrix IGF-I and TGF-beta in all compartments of the femoral bone. The association between IGF-I and TGF beta is not only temporal, but with few exceptions is also observed within the animals of each age class. These data support the hypothesis that IGF-I and TGF beta play an important role in the growth-associated accumulation of bone mass. PMID- 9868289 TI - Reconstruction of extensive long-bone defects in sheep using porous hydroxyapatite sponges. AB - The capacity of hydroxyapatite (HA) implants to support large defect repair in weight-bearing long bones of large size animals was investigated. Diaphyseal resections 3.5 cm of the tibia were performed in five adult sheep. They were substituted with HA macroporous ceramic cylinders anatomically shaped, and an external fixator was assembled. The sheep were sacrificed at 20, 40, 60, 120, and 270 days after surgery, respectively. Histology and micro X-ray study of resected implants and adjacent tissues showed proper integration of ceramic with newly formed periosteal bone as early as 20 days after surgery. In one sheep, the external fixator was removed 5 months after surgery. The animal gained the ability to walk with no functional impairment until it was sacrificed 4 months later. At this time, extensive integration of ceramic with bone was detected radiographically and confirmed by a morphological study of the resected sample. Our data indicate that large defects in a weight-bearing long bone can be repaired to the extent necessary for full functional recovery in large animals. These data set the stage for further intervention on material properties as well as for preliminary attempts to use ceramic prostheses for reconstruction of large bone defects in humans. PMID- 9868290 TI - Isolation and unambiguous synthesis of cryptolepinone: An oxidation artifact of cryptolepine PMID- 9868291 TI - [Recent advance in the thyroid testing with special reference to the gene analysis]. AB - Thyroglobulin (Tg) is a large (660 kd) homodimeric glycoprotein molecule, encoded by a gene on chromosome 8, that is secreted uniquely by thyroid follicular cells. The steps of mature dimeric Tg synthesis include folding and assembly of nascent Tg with glycosylation, in the endoplasmic reticulum (ER), and dimerization and carbohydrate modification in the Golgi apparatus, followed by incorporation into exocytotic vesicles for export into the lumen of thyroid follicles, after which thyroid peroxidase catalyses iodination of tyrosyl residues and coupling of some of them within the Tg polypeptides to form thyroid hormones (thyroxine and triiodothyronine). Here, we reviewed recent progress in the study of Tg synthesis mechanisms, especially of the function of some molecular chaperones which possibly participate in the Tg synthesis. Our recent findings indicated that Tg mutations C1263R and C1995S caused a defect in intracellular transport of Tg. The thyroid disease caused by Tg gene mutations was considered as a model of the defect in the intracellular transport of de novo synthesized protein (the ER storage disease [ERSD]). ERSDs seen in organs other than the thyroid gland are also briefly reviewed. Gene abnormalities in the other proteins in the thyroid gland, such as thyroid peroxidase, Na/I symporter, TSH receptor, thyroid transcription factor (TTF) 1, TTF 2, and PAX 8, are also discussed. PMID- 9868292 TI - [JSCP Technical Committee on Laboratory Informatics; its 10th anniversary and future perspective]. AB - The Technical Committee on Laboratory Informatics of the Japan Society of Clinical Pathology (JSCP) was established in 1988 by the late Professor Takaoki Miyati, Department of Clinical Laboratory Medicine, Yamaguchi University School of Medicine at the 35th JSCP Annual Meeting. Since then, the chairpersons have been succeeded by Prof. Nobuyoshi Matsuda (Kawasaki Medical University) and currently Prof. Masayuki Kambe (Hiroshima University School of Medicine). Open committee meetings have been held twice a year; the one during the JSCP Annual Meeting and the other independently in Spring. There have been many important subjects discussed at the past 20 meetings, including optical card utilization, laboratory information system, quality assurance in clinical laboratory, laboratory data base, outputs and presentations of laboratory results, reference values and intervals, medical support and expert systems for laboratory results, practice guidelines in laboratory medicine, laboratory support systems for medical education and research, and medical consultation in clinical laboratory. Roles of laboratory informatics will increase steadily in the circumstances of computer-multimedia networks. PMID- 9868293 TI - [Our challenge in Kagoshima University Hospital--development of more useful clinical laboratory]. AB - In 1995, we newly developed a clinical laboratory system equipped with an expert system, which was named HIPOCLATES (Hospital Intelligent POwers of Clinical Laboratory Automation Technology with Expert System). Since this system includes an expert system, we have been able to support diagnostic and therapeutic procedure in clinics and begin developing a "zonal verification method" and "early-stage DIC diagnosis support system". The former is one of the individual quality control methods and we have attempted to develop a test assurance system in HIPOCLATES using this method. The latter system is to diagnosis DIC in the early phase. Here, we introduce the detail of our project using HIPOCLATES. Following the introduction HIPOCLATES, we attempted to develop a highly integrated system in physiological examinations, and succeeded in establishment PLATON (Physiological LAboratory TOtal Network system) in 1997. Thereafter, we planned a graphic reporting system named GALIREO (Graphic Assistant Laboratory Informational REport Operating system). In GALIREO, we attempted to connect the Department of Clinical Laboratory, Ultrasonic waves test room, Fiberscope test room, Department of Pathology and Surgical center by computer network. Developing these computer network system in our clinical laboratory we want to create electronic medical record linking with THINK (HIS in our hospital). We plan to make these expert system and electronic medical record available to clinical staff via a network to realize evidence-based medicine (EBM) and utilize these to support treatment and education. PMID- 9868294 TI - [Appropriate laboratory use of microbiological testing for quality control and nosocomial infection control]. AB - Useful information from clinical microbiological tests is required for clinical diagnosis and treatment of infectious disease, whereas concern about the quality of information is still low. To provide reliable information in clinical microbiological testing, it is necessary to control the quality of clinical specimens, because of its random character. Through effective use of laboratory testing database, it will be possible to shift away our vague management of pre analytic phase of quality control so far to its established system based on objective evaluation. During the past 4 years, after the introduction of microbiological tests and information system in our hospital, the characteristics of sputum have become worse contrary to our expectations. This suggests that quality control needs successive improvements, even in service departments such as clinical laboratories. Application of laboratory testing database to nosocomial infection control is considered to be a performance of post-analytic phase of quality control from the viewpoint of effective use of laboratory data. We presented in this paper our practical application of the database to the short term and long-term control of nosocomial infection. PMID- 9868295 TI - [Communications between hospital laboratory personnel and physicians]. AB - The availability of internet facilities have been spreading rapidly and widely over the past several years. We publish a laboratory data handbook every year and distribute this to physicians in our hospital. Recently we have moved the information to our web site of the Internet. Using the web site, physicians can refer to the information on a 24-hour basis. This information includes: reference ranges; correlation of data obtained by previous and new methods; procedures for ordering each type of laboratory test. Furthermore, anyone can send questions and opinions by E-mail through the web page. The graphs which show the average values of patient results for each day can be obtained by using our database. These graphs are useful for long-range quality management of precision. Another important application is displaying histograms showing the distribution of test results between patients with a specific disease and healthy subjects. The application is expected to facilitate progress in laboratory medicine. PMID- 9868296 TI - [Computer-assisted education of laboratory medicine based on common disease database and electric RCPC system using Internet technology]. AB - We are developing computer-assisted instruction systems (CAI) for efficient education of laboratory medicine. To practice evidence-based laboratory medicine and put the education along the way, we gathered a large number of clinical records and constructed a database of clinical symptoms and signs of major diseases. We also constructed a web-based RCPC system featuring interactive multimedia materials and Q/A sessions to guide the students along important points in laboratory interpretations. We have been using these systems for 5th grade medical students and evaluated the effectiveness of them for the students to obtain knowledge regarding laboratory diagnosis. Although we still have a lot of aspects to improve, students showed interests in them and used them willingly. We believe that these CAI systems contribute to raise students' ability of problem solving in laboratory medicine. PMID- 9868297 TI - [Effective use of a laboratory database: quality assurance and laboratory workflow applications]. AB - Recent laboratory information systems have usually adopted a client server system. Computing tools which can provide easy access to a database using simple language are now strongly needed. These functions are provided in an End User Computing (EUC) system. An EUC is defined as follows: 1) General end users can easily access the database of the laboraotry system and extract objective data stored in database. 2) The extracted data will be easily converted to files that can be processed by commercially available software. In this paper, we demonstrate the examples how to use the EUC for a quality assurance system and analyses of laboratory workflow. In the case of quality assurance, we demonstrate the setting of reference intervals from stored laboratory data concerning health care examination programs at our university. Secondly, we developed a system of monitoring quality control data, and set parameters for delta checking and actual zone QC method. We can estimate and design an outline of laboratory workflow from extraction of the time currently required for each task. We can measure the turn around time for laboratory testing and rate of requests for laboratory tests received from physicians via order entry system. Moreover, we can estimate and simulate the waiting time and time required for analyses by outpatient clinics. These time monitoring systems reflect the design of laboratory workflow such as the labor and equipment time required in laboratory work. These uses of laboratory data are currently expanding further and further in the fields of education and laboratory research. We believe that information technology will facilitate future advances of laboratory medicine. PMID- 9868298 TI - [Actual situation and problems in the information office of clinical laboratories; questionnaire surveys of central laboratories of university hospitals]. AB - The laboratory tests supplied by clinical laboratory comprise an increasing volume in most hospitals. Consultation and effective utilization of laboratory data are important aspects of evidence-based medicine. Effective utilization of laboratory data will also contribute to the efficiency of hospital practice. Questionnaire surveys were conducted to investigate the actual situation in the information office of clinical laboratories in national, public and private facilities of 80 medical universities in Japan. Few facilities demonstrated efficient functioning, although information offices had been opened in six national, one public, and four private universities. The office staff received many questions on specimen handling and analytical methods. In the future, the office will be expected to be actively involved in mutual communications with clinical physicians and an information system such as computerized web is anticipated. Furthermore, a full-time laboratory physician and technicians are expected to provide support as experts in patient diagnosis. PMID- 9868299 TI - [Vascular endothelial cells and renin-angiotensin system]. AB - The vascular renin-angiotensin system (RAS) is regulated independently from circulating RAS and plays a role in the local regulation of vascular tone, the modulation of sympathetic activity and vascular remodeling. Endothelial cells are a major source of angiotensin converting enzyme (ACE), which produces angiotensin II and degrades bradykinin, in normal arteries. Mechanical stress such as transmural pressure, stretch stress and shear stress appear to contribute to the regulation of endothelial ACE activity. In contrast, vessels with intimal proliferation such as atheromatous plaque and neointima following balloon injury show expression of ACE in smooth muscle cells and macrophages in the intimal lesions. Activation of ACE in intimal SMC may relate to a phenotypic change of SMC from the contracting type of the synthetic type. Activation of ACE in macrophages is also related to the transformation of macrophages from monocytes. Concerning the role of the activated RAS, elevated blood pressure and vascular tonus by angiotensin II are candidates of vascular injury and plaque rupture. Angiotensin II stimulates migration and proliferation of smooth muscle cells and production of extracellular matrix. Furthermore, angiotensin II increases oxidized-LDL which may be related to the forming of macrophages. These evidence suggest that activation of vascular RAS following endothelial dysfunction/injury play an important role in the pathogenesis of vascular remodeling and atherosclerosis. PMID- 9868300 TI - [Endothelial expression of adhesion molecules]. AB - The adhesion molecule in the vascular endothelial cell is outlined, we reports on our result about the relation etc. among the adhesion molecule, the acute myocardial infarction, and the insulin resistance, and utility as the clinical examination is described the time. This study measured the serum concentrations of circulating adhesion molecules by sandwich enzyme-linked immunosorbent assay with the acute myocardial infarction (AMI) and the insulin resistance, and the expression of adhesion molecules on lymphocyte were analyzed by flow-cytometry. Only ELAM level in AMI disease were significantly higher (p < 0.01) than controls. sELAM has the possibility of the foresight marker of the AMI diagnosis. The correlation of SSPG value of index of the insulin resistance and IL-6, sELAM 1, and sICAM-1 takes part in condition when the immune reaction through cytokine and the adhesion molecule administered insulin. The relation between insulin and the adhesion molecule was suggested from the result of using the vascular endothelial cell. Endothelial cells are stimulated dramatically and increase the appearance of adhesion molecules on surface of the cell by the interactions with leukocytes. In these interactions with AMI and the insulin resistance, ELAM, ICAM 1 and VCAM-1 are suggested to play the most important role. PMID- 9868301 TI - [Adhesion of leukocytes to endothelial cells in atherosclerosis]. AB - The adherence of blood monocytes to the endothelium, followed by transmigration beneath the endothelium, are initiating events in the formation of foam cells, promoting atherogenesis. We showed that adhesion molecules on leukocytes were up- or down-regulated in atherosclerosis, when binding of monoclonal antibodies was measured by indirect immunofluorescence with flow cytometry. Expression of PE-CAM 1 (CD31) on monocytes and LFA-1 (CD11a) on lymphocytes was increased with age. Expression of PECAM-1 in monocytes was also up-regulated in patients with coronary artery disease. Being unchanged on aging, expression of HAR (CD44) on polymorphonuclear leukocytes and monocytes was increased in patients with coronary artery disease. On the other hand, expression of L-selectin (CD62L) on polymorphonuclear leukocytes, and LFA-1, CR3 (CD11b) and VLA-4 (CD49d) on monocytes was decreased. These findings may show the mechanism of increased chemotaxis of monocytes beneath the endothelium during the incipient stage of atherosclerosis. PMID- 9868302 TI - [Neovascularization and HGF: neovascularization in proliferative diabetic retinopathy and intraocular HGF]. AB - Human hepatocyte growth factor (hHGF) has a strong angioneogenetic action. The present study was designed to investigate the possible involvement of hHGF in neovascularization in proliferative diabetic retinopathy by measuring vitreous hHGF concentration, and to examine the gene expression of hHGF in retinal Muller cells, which are presumed to play a role in proliferative diabetic retinopathy. Patients who had undergone pars plana vitrectomy were studied (33 diabetic patients with proliferative retinopathy and 20 nondiabetic subjects). The mean vitreous hHGF concentration was higher (p < 0.0001) in diabetic subjects with proliferative retinopathy (5.7 +/- 0.7 ng/ml) than in nondiabetic subjects (1.6 +/- 0.2 ng/ml). Furthermore, diabetic subjects with iris neovascularization, which is suggestive of advanced retinal ischemia, showed higher values of mean vitreous hHGF concentration than those without iris neovascularization (7.3 +/- 1.2 ng/ml [n = 14] vs. 4.5 +/- 0.7 ng/ml [n = 19], p < 0.01). Expression of hHGF gene was detected in cultured human Muller cells. Our results indicate that hHGF may be produced in the eye by retinal cells such as Muller cells and may play a role in neovascularization of proliferative diabetic retinopathy. PMID- 9868303 TI - [Multi-parametric analysis for leukemic immunophenotyping]. AB - We have investigated whether immunophenotyping measured by laser flow cytometry could be corresponding to FAB classification of acute leukemias, using multi parametric analysis. Sixty-one patients with acute leukemias have been evaluated, including 23 patients with acute lymphocytic leukemias (ALL) and 38 patients with acute myelogenous leukemias. In principal component analysis, positive cell surface antigens could be classified into groups along to the cell lineage and differentiation. In discriminant analysis, the sensitivity by immunophenotypic method to FAB subtypes was 75%, whereas the specificity was over 90%. In cluster analysis, patients have been classified into 4 groups, which essentially corresponded to ALL, M1/M2, M3 and M4/M5. Based on those multi-parametric analysis, a new flow chart has been established, resulting that the sensitivity and the specificity was improved to over 90% and 95% respectively. These results suggest that the classification of acute leukemia using the flow chart could be useful tools for diagnosis of subtypes of acute leukemias. PMID- 9868304 TI - [The relationship between subjective sleepiness and polysomnographic findings in sleep apnea syndrome]. AB - We investigated factors of daytime sleepiness in 22 middle-aged male patients with sleep apnea syndrome (SAS) using the Epworth sleepiness scale (ESS) and polysomnography. The subjects were classified into two groups according to ESS score as follows; low ESS group: ESS score < 10, and high ESS group; ESS score > or = 10. ESS score was significantly correlated with duration in which nocturnal oxygen saturation decreased below 90% (Time of SpO2 < 90%) (r = 0.54, p < 0.05). Time of SpO2 < 90% and percent of movement arousals at the termination of apnea/hypopnea (number of movement arousal/total number of apnea/hypopneas x 100) were significantly higher in high ESS group than in low ESS group. Our findings suggest that the severity of oxygen desaturation and sleep fragmentation caused by arousal response at the termination of apnea/hypopnea may be important factors of daytime sleepiness in patients with SAS. PMID- 9868305 TI - [A case of aortic coarctation presenting with Brown-Sequard syndrome due to radicular artery aneurysm]. AB - We report a case of aortic coarctation who showed a Brown-Sequard syndrome due to the aneurysm of the anterior radicular artery. A 71-year-old right-handed hypertensive woman was admitted to the Research Institute for Brain and Blood Vessels because of subarachnoid hemorrhage in 1984. No aneurysm was detected even on the four-vessel cerebral angiography. However, the left subclavian artery was occluded associated with the coarctation. In 1997, she complained of numbness of the right lower extremity, which gradually expanded to the back and the right upper extremity. On neurological examination, she exhibited a mild weakness of the right lower extremity and a dissociated sensory disturbance below the level of C5. The cervical MRI showed a space occupying lesion at the level of C4. The right vertebral angiography revealed an aneurysm of the left anterior radicular artery at the level of C4, which compressed the spinal cord. Previously only a few cases of an aortic coarctation associated with the radicular artery aneurysm were reported in our country. The age of onset of this case was older than in the previous cases, and the location of the aneurysm was higher than those of the previous cases. Probably due to the occlusion of the left subclavian artery, the transverse collateral circulation was produced from the right to the left anterior radicular arteries at the level between C2 and C5, causing a radicular artery aneurysm in this patient. PMID- 9868306 TI - [A case of Churg-Strauss syndrome overlapping with sarcoidosis]. AB - We report a case of a 55-year-old woman with Churg-Strauss syndrome (CSS) overlapping with sarcoidosis. She had suffered from bronchial asthma since she was 42, then was diagnosed with sarcoidosis at the age of 50, with typical bilateral hilar lymphadenopathy and noncaseating granulomas in the specimen from the scalene lymph node biopsy. Five years later she was admitted to our hospital in a state of mild confusion. Since she had mononeuropathy multiplex and positive serum pANCA, in addition to bronchial asthma and marked eosinophilia, she was diagnosed as having CSS too. To the best of our knowledge, no case reports on CSS overlapping with sarcoidosis have yet been published. Although CSS can occur with sarcoidosis merely by chance, both disorders cause systemic granulomatous vasculitis and share several clinical features. Therefore, we speculate that CSS and sarcoidosis share pathogenesis in part and may present a clinical condition in which CSS overlaps with sarcoidosis. The disturbed state of consciousness in this case may be caused by a diffuse cerebral dysfunction from microangiopathy in the brain. PMID- 9868307 TI - [A case of Satoyoshi syndrome with symptoms resembling neuroleptic malignant syndrome]. AB - Satoyoshi syndrome is a rare neurological disorder of unknown etiology characterized by progressive muscle spasms, alopecia, diarrhea and skeletal abnormalities. We here describe a 25-year-old man who developed symptoms similar to neuroleptic malignant syndrome (NMS). He began to have the clinical characteristics of Satoyoshi syndrome at the age of 12 years. He was admitted to hospitals many times with painful muscle spasms and pyrexia in the early stage of the disease. He received steroid pulse therapy and oral prednisone at the age of 19, the extent and frequency of the spells being reduced thereafter. He was admitted to our hospital due to recurrence of his usual muscle spasms. He was treated with midazolam intravenously to relieve severe muscle ache, pain in the left shoulder, and insomnia. About 90 minutes later, he became comatose, with the following manifestations: hyperthermia, low blood pressure, tachycardia, profuse perspiration, acute respiratory failure, and ensuing cardiac arrest. He developed rhabdomyolysis, acute renal failure, hepatic damage, and diffuse intravascular coagulation. Serum creatine kinase level was elevated to 306,910 IU. He died of multiple organ failure 13 days after admission. His symptoms resembled NMS and malignant hyperthermia (MH). None of patients with Satoyoshi syndrome accompanied by NMS or MH have been reported. It remains to be clarified whether midazolam administration induces NMS in Satoyoshi syndrome. Nevertheless, careful attention should be paid when one administers midazolam to patients with this syndrome. PMID- 9868310 TI - [A case of juvenile muscular atrophy of the upper limb with intraspinal cavity formation]. AB - The patient was a 23-year-old man. He had no history of trauma in the head and neck. When he was 20 years old (in 1992), he noticed muscle atrophy and weakness in the right hand. In a hospital, he had cervical MRI study and other examinations. The diagnosis was juvenile muscular atrophy of the upper limb (Hirayama's disease). After that the symptoms became worse in the bilateral forearms and hands. The neurological examination showed severe atrophy in bilateral C7 to Th1 innervated muscles, right pyramidal sign, mild superficial and deep sensory disturbance in the dermatome of C7-Th1, and right Horner's sign. Cervical MRI and myelography revealed the atrophy of cervical cord and intraspinal cavity formations from C5 to Th1. We concluded that chronic and intermittent compression to cervical cord with flexion position made the cavities during the clinical course because these cavities were not found in the MRI taken in 1992. PMID- 9868309 TI - [A case of HTLV-I associated polyradiculoneuropathy and cerebral white matter lesions with smouldering type adult T cell leukemia]. AB - A 45-year-old female was admitted to our hospital complaining of abnormal sensation in the upper and lower extremities. She was given a diagnosis of smouldering type adult T cell leukemia 9 months before admission. On admission, neurological examination revealed sensory disturbance of polyneuropathy type, and generalized hyporeflexia. Nerve conduction study demonstrated decreased F wave appearance rate. Motor unit potentials in electromyelography had long duration and high amplitude. MR imaging of the brain showed multiple abnormal lesions in the periventricular and subcortical white matter. CSF protein was mildly increased, and anti-HTLV-I antibody was positive. Treatment with corticosteroid was effective, resulting in disappearance of sensory symptoms and normalization of F wave appearance. The present case is different from HAM, and the above clinical features may fit to the disease entity of HTLV-I associated polyradiculoneuropathy and cerebral white matter lesions. PMID- 9868308 TI - [A case of MELAS showing CSF pleocytosis associated with stroke-like episodes]. AB - A 55-year-old woman, who had two episodes of difficulty in putting a key into a keyhole probably due to optic ataxia at age 52 and 54 years old, developed speaking errors and was admitted to our hospital. She was 152.5 cm in height and 52.5 kg in weight. Neurological examination revealed right homonymous hemianopsia and sensory aphasia. A CSF examination revealed lymphocytic pleocytosis of 88/microliter. Serum lactate and pyruvate were remarkably increased after an aerobic exercise test. A few ragged-red fibers were present in the biopsied brachial biceps muscle. Brain MRI by FLAIR method showed scattered high signal lesions in the left temporal lobe, bilateral parieto-occipital lobes, left insular cortex and left thalamus. The left superficial temporal lesion was enhanced by gadolinium-DTPA. The proton MRS demonstrated the lactic acid peak as well as the decrease of NAA/choline ratio (0.38) in the left parieto-occipital region. Thus, she was diagnosed as a case of MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes) and successfully treated with ubidecarenone (150 mg/day). Six months later, she again developed seizure, right hemiparesis and deterioration of aphasia and presented again CSF lymphocytic pleocytoses of 15/microliter. Brain MRI demonstrated new lesions in the left temporoparietal lobes, left insular cortex and left corona radiata. Therefore, CSF pleocytosis appeared to be associated with stroke-like episodes in this case. Although the mechanism of CSF pleocytosis remains to be elucidated, it may involve the breakdown of blood-brain barrier caused by mitochondrial dysfunction. Otherwise, an inflammatory process similar to that in cases of Leber disease, who developed multiple sclerosis-like additional lesions in the central nervous system, may also take place in MELAS. PMID- 9868311 TI - [A case of acquired immune deficiency syndrome presenting acute lumbosacral polyradiculopathy due to opportunistic infection of cytomegalovirus]. AB - We reported a case of acquired immune deficiency syndrome (AIDS) with acute lumbosacral polyradiculopathy (ALSP), resulting from the opportunistic infection of cytomegalovirus (CMV). A 22-year-old Thai woman noticed weakness of the both legs, and two weeks later, she became unable to walk and had the difficulty of voiding. Neurological examination revealed flaccid paraplegia, sensory disturbance of the both legs, areflexia of the patella and ankle, and urinary retention. She could not move the legs on either side except for ankle flexion or extension, and the passive elevation of the leg brought about severe sacral pain. Radiological examinations, including lumbar MRI, failed to reveal abnormal findings. The needle EMG showed an acute denervation of the lower leg muscles, and the lumbar puncture yielded a colorless fluid containing 2,097/cu mm WBC (polynuclear 88%), 412 mg/dl protein and 45 mg/dl glucose. The serum HIV-1 antibody was positive with a marked loss of CD4 lymphocytes (31/cu mm). In CSF, the DNA of CMV was detected in the polymerase chain reaction (PCR) method. In addition, large round cells with intranuclear or cytoplasmic inclusions showing immunopositivity for the CMV antibody were present. Ganciclovir (daily dose: 400 mg, every 12 h) was administered for two weeks, but the painful numbness gradually extended to the trunk. For AIDS patients, ALSP caused by the CMV infection is a rare neurological complication, and this is the first case report in Japan. Progressive flaccid paraplegia with sensory disturbance, radicular pain, or bladder dysfunction are characteristic symptoms, and CSF pleocytosis with elevated protein or hypoglycorrhachia provides a diagnostic clue for clinicians. In addition, the CMV-DNA amplification in the PCR method or immunohistochemical approach from CSF is a useful procedure. PMID- 9868312 TI - [A case of primary Sjogren's syndrome with CNS disease mimicking chronic progressive multiple sclerosis]. AB - We report a 40-year-old woman with primary Sjogren's syndrome (SjS) with slowly progressive CNS disease. At age 38, she noticed spasticity and very gradual onset of monoparesis in the left leg. She hardly walked by herself at age 40. On admission, neurological examination revealed mild slurred speech, vertical nystagmus, spasticity in the four extremities, spastic monoparesis of the left leg, exaggerated jaw jerk, hyperreflexia in all limbs except right biceps and brachioradialis reflex, and positive bilateral Hoffmann reflexes and Babinski signs. Laboratory examinations disclosed positive anti-nuclear antibody (speckled type) and anti-SS-A/Ro antibody (64x). CSF examination revealed cell count 8/mm3, protein 42 mg/dl, 4 bands of oligoclonal band and the elevation of IgG index. MR imaging presented multiple plaque-like lesions in white matter of cerebrum and brainstem, which did not show gadolinium enhancement. Additionally she complained of dry eyes. Lacrimal and salivary secretion tests showed hyposecretion of tears and hyposialosis. The biopsied specimen of labial minor salivary gland revealed the destruction of the ducts and periductal lymphocytic infiltrations. The diagnosis of primary SjS was confirmed. We herein report a rare case of primary SjS with CNS disease mimicking chronic progressive multiple sclerosis (MS), and discuss a difficulty in differentiating CNS disease of SjS from MS. PMID- 9868313 TI - [A severe case of acute autonomic and sensory neuropathy]. AB - Acute autonomic and sensory neuropathy (AASN) is a rare neuropathy characterized by acute autonomic dysfunction and objective sensory disturbances. A 26-year-old pregnant woman with severe autonomic and sensory dysfunction is reported. This patient suddenly developed marked nausea and vomitting in about 2 days after having a sore throat. She then developed signs of autonomic dysfunction including dilated non-reactive pupils, dryness of the eyes and oral mucous membranes, generalized anhidrosis, paralytic ileus, orthostatic hypotension, and continuous tachycardia. She also had severe generalized sensory impairments of all modalities, and all deep tendon reflexes were absent. Sensation was almost totally lost for all modalities below the neck. There was marked pseudoathetosis and sensory ataxia in all extremities. Motor examination was normal. She had inability to urinate. At this time she was 38 weeks pregnant, and when she showed signs of fetal distress, a Caesarean section was performed. Albumino-cytological dissociation was seen in the CSF. Serum noradrenaline was reduced, no sensory nerve action potentials could be elicited, and reduced coefficient of variation of the R-R interval on electrocardiography was observed. Plasma exchange was performed every other day for 3 days for about 3 weeks after the onset of the illness, but no favorable effects. Seven months after the onset, her autonomic dysfunction slightly improved, but there was no recovery from the sensory disturbances. Many symptoms and signs that characterize AASN occurred in this patient, and each was severe. The patient developed SIADH, sleep apnea, personality change, and amenorrhea in the course of the disease. We suggest that AASN patients might have both peripheral and central nervous system manifestations including seizures and personality changes. PMID- 9868314 TI - [Two cases of neuralgic amyotrophy]. AB - Case 1: A 27-year-old man had a fever of 38 degrees C, followed by acute onset of bilateral upper arm pain. Two days later severe muscle weakness in bilateral upper arms appeared and he was admitted to our hospital. On admission, severe atrophy of the left deltoid and mild atrophy of the right deltoid were observed, with severe muscle weakness in bilateral deltoid and mild weakness in other parts of upper extremities. Tendon reflexes were decreased in the upper extremities. Sensation was intact. CSF showed mild pleocytosis. Nerve conduction velocity was normal and electromyography showed mild NMU decrease in upper extremities. Muscle biopsy of the right deltoid one month after the onset was normal. Muscle weakness began to improve 3 months after the onset, with only mild weakness at 10 months. Case 2: A 60-year-old man had acute onset of left shoulder and upper arm pain, followed by muscle atrophy and weakness of the left upper arm. He showed marked atrophy of the left deltoid, moderate atrophy of the left biceps and left scapular region, and severe muscle weakness in the left upper arm. Deep tendon reflexes were absent in the left upper extremity. Sensation was intact. Nerve conduction velocity was normal and electromyography showed marked NMU decrease in the left upper arm. Muscle biopsy of the left biceps 4 months after the onset showed grouped atrophies on HE staining, type 2 fiber atrophies on routine ATPase staining, and many targetoid atrophic fibers on NADH-TR staining. Muscle weakness began to improve slowly 6 months after the onset, but considerable weakness persisted at 10 months. Detailed muscle biopsy findings in neuralgic amyotrophy have not been documented. Muscle biopsy of Case 2 showed marked neurogenic changes compared to Case 1, which may be associated with the difference in clinical course between the two cases. PMID- 9868315 TI - [An autopsied case of multiple system atrophy with remarkable cerebral atrophy]. AB - In this paper, we report the pathological findings of a multiple system atrophy (MSA) patient with remarkable frontal lobe atrophy. The patient was a 65-year-old woman with a 13-year history of untreatable parkinsonism, dysautonomia and progressive motor aphasia. The cranial imaging study disclosed progressive atrophy of the frontal lobe, striatum and pontocerebellar system. She died of pneumonia. The brain weight was 810 g, and there was remarkable atrophy of the cerebrum predominantly in the frontal lobe, striatum, pons and cerebellum. The microscopic examination revealed a preserved cortical structure with laminar gliosis in the sixth layer of the precentral and superior frontal gyri of the frontal lobe, and postcentral gyrus and inferior parietal lobule of the parietal lobe. The cortices of these regions also revealed to be in a spongy state in the second layer, and mild cell loss was seen in the fifth and six layers. The frontal lobe white matter showed a slight pallor, a mild loss of myelinated fiber and axon, and mild gliosis. Glial cytoplasmic inclusions (GCIs) were abundantly observed in the deep layer of the cortex in the regions mentioned above, and were more abundant in the white matter of the frontal and parietal lobes, callosal body, and internal, external and extreme capsules. There were severe degenerations in the olivopontocerebellar and striatonigral systems, and GCIs in widespread regions of the brain. We could not detect any Pick bodies, Lewy bodies, ballooned neurons, senile plaques, or any significant amounts of neurofibrillary tangles. There were no vascular changes. Thus, this was a verified MSA patient with progressive aphasia and remarkable frontal lobe atrophy. We indicate a possible involvement of the cerebral lobes in MSA. PMID- 9868316 TI - [A case report of steroid-responsive progressive encephalomyelitis with rigidity showing muscle stiffness limited to the upper body]. AB - A 37-year-old female presented with progressive encephalomyelitis with rigidity for a period of about three months. Spasms began with several minutes of paroxysmal painful muscle stiffness in the left upper limb, followed by pain and muscle spasms in the upper limbs, shoulders, neck and back. The paroxysmal painful spasms were easily evoked by light touches and startling. The patient had contracture in the upper limbs and difficulty in relaxing the muscles. She also developed brain stem signs such as left sixth nerve palsy. The serum antinuclear antibody was positive (1 : 160). The cerebrospinal fluid examination revealed mild pleocytosis and protein elevation. Brainstem MRI, cervical MRI, and electroencephalography were normal. Surface electromyography in the arm and neck muscles showed continuous motor unit discharge elicited by passive movement of the right arm. Diazepam and baclofen were minimally beneficial. After treatment with intravenous injection of high-dose methylprednisolone and sequential oral prednisolone administration, the patient showed substantial improvement. This disorder may be part of a clinical spectrum that has an underlying autoimmune basis. PMID- 9868317 TI - [Exercises on a bicycle ergometer in a family of diabetes mellitus associated with a mutation of mitochondrial DNA]. AB - The serum pyruvate and lactate levels were studied after exercise on a bicycle ergometer in a family of diabetes mellitus (DM) associated with a mutation at nucleotide 3243 in the mitochondrial gene. A 56-year-old Japanese woman with the mutation at a percentage of 5% in the blood had insulin-dependent DM and sensory hearing loss without muscle symptoms. Her serum lactate and pyruvate levels increased markedly during and after exercise on a bicycle ergometer. Two of her sons were found to have the same mutation at a percentage of 17% and 18%, respectively. Her 26-year-old son was found to have borderline DM after oral glucose loading, although he showed no abnormalities of the metabolism of pyruvate and lactate. Her 31-year-old son showed no abnormalities after oral glucose loading and after exercise on a bicycle ergometer. Although the same mutation causes more severe MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes), little is known about whether these diabetic patients are subclinically involved with myopathy. The noninvasive ergometer exercise with determination of serum pyruvate and lactate may be useful in evaluating the severity of myopathy in these patients. PMID- 9868319 TI - [Midbrain infarction presenting with unilateral blepharoptosis, trochlear nerve paresis and MLF syndrome]. AB - We report a 75-year-old hypertensive man presented with unilateral MLF syndrome combined with ipsilateral blepharoptosis and trochlear nerve paresis due to midbrain infarction. He was admitted to our hospital for sudden onset of diplopia. Neurological examination revealed left ptosis, infraduction disorder of the left eye and left MLF syndrome. Horner's syndrome or other focal neurological signs were not observed; convergence was preserved. Hess-charts confirmed left superior oblique paresis and left internuclear ophthalmoplegia. Laboratory data were all normal. T2-weighted images of brain MRIs demonstrated a high-signal intensity lesion in the paramedian dorsal midbrain at the inferior colliculus level, anterior to the cerebral aqueduct. The oculomotor dysfunction and diplopia had disappeared at discharge, although slight ptosis persisted. It is likely that the lesion in our case affected the left MLF, the right trochlear nucleus or its fascicles and the left partial oculomotor fascicles. Cases with MLF syndrome associated with trochlear nerve palsy have been rarely reported. Our case suggests that the fibers controlling for the musculus levator palpebrae superioris are located more caudally in the oculomotor fascicles. PMID- 9868318 TI - [Monoparesis of the left leg with bilateral pyramidal signs due to posterior spinal artery syndrome]. AB - A 63-year-old hypertensive and diabetic woman suddenly experienced severe pain in the back and abdomen of the left side. Sensory disturbance and muscle weakness in her left leg and urinary retention followed. Somatosensory evoked potential (SEP) indicated a thoracic cord lesion. Furthermore, MRI showed a focal lesion with a high signal intensity on T2-weighted images in the left posterolateral region of the cord Th4-9. She was diagnosed with posterior spinal artery syndrome (PSAS) due to spinal infarction by clinical presentation and MRI findings. Monoparesis can be caused by PSAS. PMID- 9868321 TI - [Beneficial effects of acetazolamide on paroxysmal attacks of girdle sensation in multiple sclerosis]. AB - A 56-year-old woman with a 40-year history of multiple sclerosis (MS) developed paroxysmal attacks of girdle sensation in the Th5-6 dermatomes. The attacks lasted 20-60 minutes and occured up to three times per week. T2-weighted MR imaging of the spinal cord showed high intensity area from Th5 to Th8. Electrocardiography, echocardiography and laboratory findings did not indicate ischemic heart disease; therefore, the paroxysms were attributed to the spinal cord lesions. Attacks were successfully suppressed by acetazolamide 250 mg/day. Although carbamazepine is frequently used to treat paroxysmal attacks in MS, we would like to suggest that acetazolamide may also be beneficial in some patients with paroxysmal symptoms. PMID- 9868322 TI - [Importance of the study of chromosomal alterations in acute leukemias]. PMID- 9868320 TI - [A case of thoracic extradural spinal cord teratoma with neurological sequelae more than 10 years after surgery]. AB - We describe a rare case of a 42-year-old man with extradural spinal cord teratoma who presented with left Brown-Sequard syndrome more than 10 years after surgery for a left posterior mediastinal tumor. When he was 29 years old, he underwent an operation for a posterior mediastinal tumor. Histological findings showed adenocarcinoma in teratoma. Ten years after the operation, the patient developed left leg weakness. Three years later, he developed superficial sensory disturbance in his right leg. MRI showed extradural tumor formation at the level of the Th 9 vertebra, which compressed the spinal cord from the left posterior side. At surgery, the tumor was well encapsulated and extended to the left posterior side of the spinal cord through the intervertebral foramen. The histological findings were almost identical to those from the mediastinal tumor. This dumbbell tumor apparently developed from the remainder of the posterior mediastinal tumor over a period of more than 10 years. PMID- 9868323 TI - Antithrombin, protein S and protein C and antiphospholipid antibodies in systemic lupus erythematosus. AB - Patients with systemic lupus erythematosus (SLE) have an increased risk of thrombosis, related to the lupus anticoagulant or anticardiolipin antibodies (ACL). Antithrombin (AT), protein C (PC) and protein S are natural anticoagulants with a very important role in the regulation of coagulation and fibrinolysis, and in the prevention of thrombosis. Total protein S (TPS), free protein S (FPS), AT and PC were measured in 53 unselected patients with SLE in order to verify their relation with previous history of thrombosis or the presence of anticardiolipin antibodies (ACL). The influence of prednisone use on the concentration of these natural anticoagulants was also analysed. The control group was formed by 20 blood donors. The PC concentration was increased in patients with ACL, when compared to controls (p < 0.01), or patients without ACL (p = 0.01). FPS levels were decreased in patients with SLE in relation to controls (p = 0.01), but not related to thrombosis or ACL. There was no difference in plasmatic concentration of AT and TPS, between patients and controls. PC, AT and TPS were increased in patients under prednisone use (p < 0.01). There was no association between concentration of natural anticoagulants and a history of thrombosis. Although decreased FPS was found, or results suggest that it seems not to be a risk factor for thrombosis in SLE. Since PC and AT were not decreased, they should not be involved with thromboembolic complications in these patients. The use of prednisone can influence concentration of natural anticoagulants in patients with SLE. PMID- 9868324 TI - [Assessment of blood volume in children by means of different prediction formulas]. AB - PURPOSE: To assess the blood volume (BV) of children by using different prediction formulas (PF) and to confirm the value of these in providing reference estimates. PATIENTS AND METHODS: The results of BV measurements in 19 children, performed at the Department of Physiology of the Haematology and Immunology Institute, were studied. The children had been sent to this centre to define the diagnosis of anaemia (11 cases) or polycythaemia (8 cases). Four adult and 2 children formulas based on body measurements were applied in order to estimate the reference BV. The relationship between the measured red cell volume (RCV) and plasma volume (PV) and the reference values was calculated for each formula, and the results were evaluated by establishing the normality criteria. RESULTS: The PF used provided reference estimates of the different BV for a given patient. It was found that most adult formulas gave RCV and PV estimates which differ from those of children. PV was commonly decreased with all the formulas. CONCLUSIONS: The different PF provide distinct BV estimates in most patients, and coincidentally the results differ in any given child. The formulas applied to adults, based on weight and height or body surface, should not be applied to estimate the BV in children. The use of equations based on height is recommended, correlating the BV assessment with the results of other studies. PMID- 9868325 TI - [Which are the true incidence rates of primary hematological disorders acquired in our population?]. AB - PURPOSE: To define demographic and epidemiological characteristics of primary haematological disorders (PMHD) in patients referred to a haematology department in 1,240 beds general hospital. PATIENTS AND METHODS: PERIOD OF STUDY: 01/94 12/94. We have performed a study in patients older than 14, to determine the age adjusted incidence rates of PHD in the assigned population: 439,279 inh (M: 210,139; F: 229,140), with a negative-vegetative growth (-1.39/10(3) inh/y). A total of 1,242 new cases was received, 302 of them were diagnosed of PHD. Epidemiological method: incidence rates (IR), age and sex adjusted incidence rates (AIR), truncated standardized incidence rates (TSIR) and confidence interval (CI) were calculated. STATISTICAL METHOD: Normal distribution, descriptive and frequency distribution analysis were performed along with chi 2 test. RESULTS: Demographic data: mean age (+/- SD): 63.54 y +/- 15.81; range 19 92. M/F: 177/125, males mean age 62.85 +/- 16.29, females: 64.52 +/- 15.11. The PHD distribution was: MGUS 84 cases; NHL 57; MDS 33; CLL 26; CMPD 26; MM 21; HD 14; AL 11; ITP 10; CML 9; AIHA 5; hypoplastic anaemia 3; and cryoglobulinaemia 3. AIR (cases/10(5) inhab/y): Consulting rate 261.79. The PHD incidence rate was 31.00 (M: 38.01; F: 25.51). In patients under 60 y the AIR of PMDH was 31.31 (M: 36.42, F: 26.25), and in those older than 60 y the AIR was increasing, with 178.86 (M: 247.21, F: 128.52). The AIR for subtypes was: MGUS, 8.01 (M: 3.56, F: 1.93); CLL 2.28 (M: 2.69, F: 1.95); CMPD 3.17 (M: 2.83, F: 3.51); MM 1.92 (M: 2.43, F: 1.49); HD 2.27 (M: 3.54, F: 1.04); AL 1.41 (M: 1.54, F: 1.30); ITP 1.15 (M: 1.20, F: 1.12); CML 1.09 (M: 1.19, F: 1.01); AIHA 0.61 (M: 0.55, F: 0.69); hypoplastic anaemia 0.24 (M: 0.19, F: 0.30); cryoglobulinaemia 0.34 (M: 0.55, F: 0.14). REMARKS: The elderly have increased incidence of PHD. The AIR is higher in males and in older than 60 y, unless for CMPD. Most frequent PHD were MGUS and NHL. PMID- 9868326 TI - [Complications of plateletpheresis procedures]. AB - PURPOSE: Thrombopheresis procedures have been recently expanded with the development or different programmes. Taking into account that this reasonably safe procedure is not devoid of complications, it would be desirable to select those individuals with lower risk of suffering adverse side effects as donors. The thrombopheresis procedures performed in our hospital between 1986 and 1997 were analysed in order to establish the useful guidelines for such selection. MATERIAL AND METHODS: All the thrombopheresis procedures performed in the Asturias Central Hospital blood bank in the 1986-1987 period were analysed. The first procedure per donor, along with all data referred to adverse effects appearing during thrombopheresis, were collected. Sex, age, body, weight, blood cells count (before and after thrombopheresis) and serum calcium levels (before and after thrombopheresis) were taken as variables with predictive value for adverse effects. With regard to the procedure, the model of cell separator, the duration of the procedure, the amount and type of anticoagulant solution and the prophylactic use of calcium ions were assessed. RESULTS: A total number of 1,024 thrombophereses were analysed. Some types of adverse effect were seen in 259 instances (25.3%). Of these, 70.3%, were mild, 29.3% moderate and 0.4% severe. The commonest adverse effect was perioral paraesthesia. Of the different variables studied, female sex and low weight acquired predictive value with respect to the occurrence of adverse effects. Prophylactic administration of calcium did not prevent the appearance of complications. CONCLUSIONS: The thrombopheresis procedures may present adverse side effects in a high percentage of cases, which, although mostly mild, require specialised personnel for identification and management. Males weighing over 70 kg are less prone to suffer such effects. Oral administration of calcium before the apheresis does not prevent the adverse reactions. PMID- 9868327 TI - [Detection of parvovirus B19 in patients of the Zulia State blood bank with different hematological alterations]. AB - PURPOSE: To assess the seroprevalence of IgG and IgM specific for parvovirus B19 (B19) and its association with aplastic crisis developing in patients with different haematological disorders. PATIENTS AND METHODS: Fifty-three serum samples were evaluated, 24 from patients with aplastic crisis and 29 from others without such crises, all of them suffering from different haematological diseases diagnosed at the University Hospital of Maracaibo and the Zulia State Blood Bank, in Venezuela; 15 samples from healthy blood donors were examined as well. Indirect immunofluorescence technique was used in the study. Lymphocyte subsets were quantified in 10 of the patients with aplastic crisis by means of cytofluorometry. Serum proteins were assessed by electrophoresis in all samples. The statistical analysis was performed according to Student's t test and chi square, by applying the statix 4.0 and SAS programmes. RESULTS: Positive IgG was found in 20 of the 24 patients with aplastic crisis (83.3%), 20 of the 29 without crisis (68.9%) and 7 of the 15 healthy controls (46.6%). Positive IgM was found only in 2 of the 24 patients with aplastic crisis (8.3%). Both the patients without aplastic crises and the control groups were negative for PB19 IgM. The average CD4 and CD8 T lymphocyte count and the CD4-CD8 index in the patients studied were 454 CD4 cells/microL, 1,006 CD8 cells/microL and 0.5%, significantly different from the control group, whose figures were 860 CD4 cells/microL, 546 CD8 cells/microL and 1.6%. The average B lymphocyte count of the patients (628 cells/microL) was higher than that of the control group (349 cells/microL). The average NK cell count in the patient was 174 cells/microL, slightly below the control group (221 cells/microL). Mild beta-globulin decrease was found in the electrophoretic study of the serum proteins of the patients, along with significant increase of the total protein and the gammaglobulin fraction with regard to the control group. CONCLUSIONS: Higher PB19 IgG seropositivity was seen in the patients with aplastic crisis with respect to the control group, suggesting wider exposition to the virus among them with regard to the healthy population. Specific PB19 IgM was detected in 2 patients with immunodeficiency and aplastic crisis. A significant decrease of the CD4 and CD8 T-lymphocyte subsets, along with decreased CD4-CD8 quotient, were found in the aplastic crisis group, and an impairment of the immune response to the viral challenge can be inferred form this. The alterations of the serum proteins, together with the increased B lymphocytes, might suggest a polyclonal activation of these cells. The absence of other known lymphotropic viruses in most of the patients studied (50) show that the alterations found here are related to recent or past B19 infection. PMID- 9868328 TI - [Hypercoagulability status previous to total hip and knee arthroplasty: the contribution of rheumatoid arthritis]. AB - PURPOSE: Starting from a status hypercoagulability previous to substitutive hip and knee surgery, the aim of this work was to investigate the influence of different osteoarthropatic pictures for which arthroplasty is indicated in the activation of the clotting cascade, rheumatoid arthritis (RA) being one of such pictures. PATIENTS AND METHODS: Of 79 patients suitable for prosthetic surgery of hip (53) and knee (26), the preoperative values of several markers, namely, D dimers (D-D), thrombin-antithrombin (TAT) complex, and F1 + 2 prothrombin fragment (F1 + F2) were assessed by enzymoimmunoasay. The mean age of the patients was 65.5 years, and their sex distribution was 50 women and 29 men. The indications for arthroplasty were as follows: osteoarthrosis (62), aseptic necrosis (11), RA (9), articular gout (2), previous fracture (2), more than one diagnosis overlapped in some cases. The results attained were compared with a control group comprised of 33 subjects (16 women and 17 men) with mean age similar to the patient's group (68.06 years). RESULTS: The D-D values in the patients suitable for hip arthroplasty and the TAT values in patients suitable for both types of surgery were significantly higher than those found in the control group (p = 0.012 and 0.01, respectively). The preoperative TAT levels of the RA patients were significantly higher (p = 0.025) than those found in the patients with the other surgical indications. CONCLUSIONS: Previously to the performance of arthroplasty, the patients show hypercoagulative marker values higher than those of age-matched controls. The significant rising of TAT found in RA patients is concordant with the literature, and this fact makes it advisable to include RA among the pathologic situations associated with hypercoagulability, as this is a common indication for substitutive hip and knee surgery with high risk of venous thromboembolic disease. PMID- 9868329 TI - [Lymphocyte subsets in blood donors]. AB - OBJECTIVES: To establish the normal values among the different lymphocyte subsets in peripheral blood, measured by surface antigen expression, in healthy population. To observe if there are potential differences in respect of age, gender or sample origin. To compare the absolute lymphocyte number obtained with flow cytometry and with a cellular counter. MATERIAL AND METHODS: Longitudinal and prospective study performed with 100 samples of blood donors. Direct immunofluorescence with triple color staining was made on whole blood, red blood cells were then lysed and samples were analysed with a flow cytometer. The lymphocyte subsets studied were the T lymphocytes and their subsets (CD4 and CD8 lymphocytes), the B lymphocytes and Natural Killer cells population. The absolute lymphocyte count was performed with an automatic cellular counter. Donor data such as age, gender and origin were recorded and were statistically analysed. RESULTS: Normal ranges from the studied parameters are similar to other series. Comparison with gender, origin or age groups gave no significant difference, although there seems to be a tendency to decrease with the ageing of the population. Total absolute lymphocyte number did not differ between the results from the cellular counter or from the flow cytometer. CONCLUSIONS: We found no differences in the absolute lymphocyte number nor in the lymphocyte subsets studied (T lymphocytes, CD4 lymphocytes, CD8 lymphocytes, B lymphocytes and NK cells) with respect to gender, age or sample source. We have established reference ranges for our laboratory. We have not found significant differences in the absolute lymphocyte number measured with cell counter or with flow cytometer. PMID- 9868330 TI - [Serum hepatitis B markers in blood donors in Venezuela. What do they mean?]. AB - PURPOSE: The prevalence of serum BH markers is high in our country. In order to assess the factors of impact in this fact a group of blood donors with positive BH markers attending our Service between 1-1-95 and 12-31-96 was evaluated, although the endemic character of the disease may be an obstacle for its recognition. The residual risk of HBV transmission by seronegative transfusions was also evaluated. MATERIAL AND METHODS: The evaluation of donors was performed by surveying identification data, type and frequency of donations and risk factors. The following reagents were used to carry out the bests: Biotest Elisa HbsAgand Heprofile anti HBc, ADI Diagnostics (1995), and Abbott Auzyme Monoclonal and Abbott Corzyme Diagnostic kit. Positive donors were re-evaluated to determine any progression of the disease and they were classified into false positive (FP), chronic carriers (CC), acute hepatitis (AH), naturally immunized (NI), or without evidence of immunity (WI). The residual transfusional risk was calculated by multiplying the adjusted prevalence of HbsAg positive donors by the window period expressed in fractions of a year. RESULTS: Out of 53,338 donations, 5.91% were discarded due to the presence of HBV, 7.5% of them showing significant association with other markers: HCV, Chagas' disease and syphilis. Of all donors 223 (7.54%) returned for re-evaluation, and only 139 (62.33% of them) could be fully evaluated as follows: FP 10.09%, AH 5.76%, CC 7.19%, NI 68.35%, WI 7.91%. Their mean age was 35.73 +/- 9.95 years, and there were 112 males and 12 females. The donations were patient-related in 93.5% of cases, voluntary in 3.22% and coming from mobile units in 3.22% of instances (p = 0.067). Fifth per cent donated for the first time, and the other 50% had donated previously although not in a consecutive way (p = 0.96): 46% were ignorant of their post-donation findings, 37.09% were seemingly negative and 16.12% had positive tests. Of these last, the reasons for donation included defective questioning and information (50%), results not verified (20%) and family/hospital pressure (30%). Homo/bisexuality and drug abuase (not intravenous) were found significant when associated to each other or to promiscuity. Promiscuity attained significant risk value, especially when previous venereal disease or close contact with BH patients were presented (p = 0.0015, p = 0.155, and p = 0.0036, respectively). The residual risk of transmitting HBV by seronegative transfusions was found to be: 1:437. CONCLUSIONS: 1. No risk factors were detected in a half of the cases. 2. The other half, comprising the known seropositive subjects and the promiscuous, homo/bisexual donors and those having close contact with HBV patients, can be discarded predonation. 3. Drug abuse (not intravenous), frequently associated with other risk factors, should discard a donor. 4. History of veneral disease should be considered a risk factor due to its common association to HBV. 5. The residual risk of acquiring HBV post-transfusion is still high. RECOMMENDATIONS: 1. To improve the information provided to donors regarding the risk factors. 2. The interrogation should be simple, direct and deep. 3. To insist in the importance of the donor returning for his/her credentials in the blood bank and to know about any altered results, and to seek medical advice when appropriate. 4. To change related donation into a voluntary and consecutive pattern. PMID- 9868331 TI - [Molecular characterization of thalassemias in the Valencia community and its relationship with the hematological phenotype]. AB - PURPOSE: The aim of the study is to present the first results of molecular characterization of thalassaemias in Valencian Community and their relationship with the haematological parameters. PATIENTS AND METHODS: The study includes 87 thalassemic patients: 30 alpha-thalassaemias, 40 beta-thalassaemias and 17 delta beta-thalassaemias. The molecular alterations were studied in white cell blood DNA, either following different PCR methods or by testing the digestion of the amplified PCR products with selective restriction enzymes. RESULTS: The molecular characterization of beta-thalassaemias was achieved in 94% of the subjects, being the transition C-->T in CD-39 the most frequent (44%) of the mutations studied. 94% of the delta beta-thalassaemias studied corresponded to the delta beta Spanish type. All the alpha thalassaemias characterized (64%) corresponded to the -alpha 3.7 deletion. The reamining 36% were negative for the alpha 0 deletions - MED, --20.5, or the non deletional mutations Hph I and NocI. DISCUSSION: In the Valencian Community, like what has been described for the beta-thalassaemias in other Mediterranean regions of Spain (Barcelona, Granada and Mallorca), a high incidence in C-->T transition in CD-39 was observed, in contrast with central and south-western regions of Spain, where the G-->A IVS-I-1 is the most frequent mutation. Our study supports that the IVS-I-6 mutations is the one with lower repercussions on the haematological parameters. Our study confirms the Spanish type of delta beta-thalassaemia as the most frequent in the Valencian Community, and that the 3.7 kb alpha deletion is the most frequent mutation for the alpha thalassaemia, although alpha thalassaemia is also the poorly characterized form of thalassaemia. PMID- 9868332 TI - [Obstetric treatment of hemophilia carriers]. PMID- 9868333 TI - [Mechanisms of resistance to cytotoxic therapy]. PMID- 9868334 TI - [Evaluation and usefulness of hypercoagulability markers in medical pathology]. PMID- 9868335 TI - [Application of modified comparative genomic hybridization in the genetic analysis of 5 lymphoid neoplasms]. AB - To analyze the utility of performing modified comparative genomic hybridization (mCGH) as a complementary technique to conventional cytogenetic techniques in the genetic analysis of lymphoid neoplasms. Modified comparative genomic hybridization and subsequent FISH techniques were performed in 5 lymphoid neoplasms cases diagnosed in Fundacion Jimenez Diaz. The latter was done in order to confirm the results obtained with mCGH. Gains of chromosomal regions not detected with conventional cytogenetic techniques were detected by mCGH. A good correlation in the results obtained between conventional cytogenetic and mCGH techniques was observed. Nevertheless, mCGH enables the detection and subsequent identification of gains of genetic sequences undetectable with cytogenetic techniques with possible diagnostic and prognostic value. PMID- 9868336 TI - [Chediak-Higashi syndrome. A laboratory finding]. AB - The Chediak Higashi Syndrome (CHS) is an inherited autosomic recessive immunodeficiency rarely reported. Two pediatric cases are presented, where the first approach to diagnosis was the laboratory report of giant granulation in granulocytes and lymphocytes, observed in peripheral blood smear. In order to confirm the diagnosis of CHS, immunologic tests, skin biopsy, bone marrow aspirate and microscopic hair examination were performed. It is remarked the importance of the careful examination of the blood smear in the detection of this cases, whereas clinic manifestations are not relevant until the hematologic suspicious of the syndrome is evident. The early detection of these patients can lead to bone marrow transplant, which is the only curative treatment to this disorder, lethal in the first decade of life. PMID- 9868337 TI - [Basophilic blast crisis in chronic myeloid leukemia]. AB - Acute basophilic leukaemia is usually characterized by a very rapid clinical course, hyperhistaminemia, resistance to antineoplastic therapy and early death due to complications related to disease. This entity is a rare condition, accounting for less than two percent of all haematopoietic malignancies. Most of the case reports are basophilic blast crisis in patients with a previous lympho or myeloproliferative disorder. A 62-year-old woman who was diagnosed as Philadelphia positive chronic myelogenous leukaemia after four years of evolution developed a basophilic blast crisis, whose characteristics are reported. Accompanying this transformation there was also a cytogenetic change. Despite chemotherapy the patient died of disease progression. PMID- 9868338 TI - [Interstitial 9q deletion: a primary change in acute non-lymphoblastic leukemia?]. AB - Two cases of acute non-lymphoblastic leukaemia (ANLL), subtypes M1 and M5b, are presented. Both of them showed the 9q-interstitial deletion as the only abnormality. From our observations and others found in the literature it can be inferred that such abnormality might be a primary finding in ANLL, related to an unfavourable prognosis, regardless the morphological subtype. PMID- 9868339 TI - [Drug-induced agranulocytosis: clinical study of 19 cases]. AB - Agranulocytosis is one of the most serious side effects to drugs. From January 1991 to June 1996 were diagnosed 19 cases of agranulocytosis associated with drugs at our hospital (incidence rate: 9.4 over million hab. per year). The average age was 62 and 11 cases were women. The drugs most commonly involved were metamizol and ticlopidine. In 15 of the patients fever blew up and 16 presented some infectious location. In 9 of the cases some positive microbiological culture was obtained, gram-negative bacilli being the commonest. G-CSF was used in 13 of the patients, observing a quicker haematological recovery (5.7 days vs 9.1, p = 0.07), though without any difference in mortality, which was of 0%. All this leads to the following conclusions: a high incidence of agranulocytosis in our environment and the important role of metamizol and ticlopidine in its origin. PMID- 9868340 TI - [Comparative study of the analyzers L-9100 (Merck-Hitachi) and HA-8121 (Menarini) for the determination of glycosylated hemoglobin]. AB - A comparison study about the determination of HbA1c by L-9100 (Merck Hitachi) and HA-8121 (Menarini) was made. HbA1c was measured by high pressure liquid chromatography (HPLC). The within-run imprecision was CV < 3% (for HA-8121) and CV < 4.9% (for L-9100). The between-run imprecision was CV < 1% (for L-9100) and CV < 2% (for HA-121). Analysis of means (Student test) showed that both instruments were statistically different for HbA1Ac measurements (p < 0.001). The results of regression analysis (Passsing-Bablock test) were as follows: y = 0.88x + 0.58 (y = L-9100, r = 0.982, p = 0.005). In conclusion, the results showed that there are statistical differences between both instruments. Regression and correlation was good, but the data also showed that there is a constant and proportional error. We must make a new reference range values. PMID- 9868341 TI - [Treatment with lymphoblastoid alpha interferon in patients with chronic myeloid leukemia refractory to recombinant interferon alpha 2]. AB - The results of the treatment with lymphoblastoid alpha interferon (IFN-alpha N1) in 10 patients with chronic myeloid leukaemia who had poor response to previous recombinant alpha interferon (rIFN-alpha) are presented. Eight of these patients had not developed anti-alpha 2 interferon antibodies, and 2 had non-neutralising anti-IFN antibodies. Three of the 10 patients received benefit from IFN-alpha N1 treatment. Two of them, with no response to rIFN alpha 2, attained complete haematologic response wit IFN-alpha N1. Cytogenetic responses although minimal, were achieved as well. The third patient, after receiving rIFN-alpha for 3 years with no response, had partial cytogenetic response after 4 months of treatment with IFN-alpha N1. These results suggest that IFN-alpha N1 when used in patients refractory to IFN-alpha N1. These results suggest that IFN-alpha N1 when used in patients refractory to IFN alpha 2 without anti-IFn alpha 2 neutralising antibodies may be useful in a minority of patients, although the frequency of cytogenetic responses is low. PMID- 9868342 TI - [Anatomy of the eukaryotic gene]. PMID- 9868343 TI - [Criteria and algorithms for the computerized validation of the hematology laboratory]. PMID- 9868344 TI - [Quarantine and plasmapheresis in a community hospital]. PMID- 9868345 TI - [Germicide metabolites derived from oxygen and ultraviolet radiation in the phagosome of polymorphonuclear leukocyte neutrophils]. PMID- 9868346 TI - [Benign medullar oligoblastosis secondary to infectious mononucleosis]. PMID- 9868347 TI - [Sideroblastic refractory anemia and acute leukemia in Waldenstrom's macroglobulinemia]. PMID- 9868348 TI - [Essential thrombocythemia and pregnancy]. PMID- 9868349 TI - [Severe peripheral blood erythroblastosis following a splenectomy in a patient with heterozygous beta thalassemia and triple alpha gene]. PMID- 9868350 TI - [Association of multiple myeloma and autoimmune hemolytic anemia]. PMID- 9868351 TI - [Hemolytic anemia due to prosthetic mechanical valves]. PMID- 9868352 TI - [Mechanisms of eosinophil development]. PMID- 9868353 TI - [Role of DARPP-32 in dopamine signal transduction in the striatum]. PMID- 9868354 TI - [The roles of chitin synthases and chitinases on the tip growth and the differentiation of filamentous fungi]. PMID- 9868355 TI - [Improvement of polyacrylamide gel media for electrophoresis: enhancement of elasticity by polymer blending]. PMID- 9868356 TI - [Two recent topics in ribosome research]. PMID- 9868357 TI - [Proteomics in pharmaceutical industry]. PMID- 9868358 TI - [Saturation mutagenesis in mice]. PMID- 9868359 TI - [Sedimentation equilibrium]. PMID- 9868360 TI - Kissing and RNA stability in antisense control of plasmid replication. PMID- 9868361 TI - Plant disease-resistance proteins and the gene-for-gene concept. PMID- 9868362 TI - Patch engineering: a general approach for creating proteins that have new binding activities. AB - Patch engineering is a technique for creating folded proteins that have new binding activities. Different protein scaffolds are used to present a patch of discontinuous residues on a folded-protein surface. By varying simultaneously the residues in these patches and displaying these mutant proteins on phage, one can select proteins that have new binding activities. Patch engineering is applicable to any protein fold. Novel proteins derived by this approach might replace antibodies in certain applications or provide lead molecules for the design of non-peptide analogues. PMID- 9868363 TI - How regulated protein translocation can produce switch-like responses. AB - It is widely appreciated that the regulated translocation of signaling proteins can increase the specificity and speed of signal transduction. It is less obvious that regulated translocation can also, in principle, turn a graded response into a more switch-like one. For example, if two or more signaling proteins are induced to translocate, the result can be a switch-like, ultrasensitive response. A switch-like response will also occur if translocation raises the local concentration of a signaling protein sufficiently to partially saturate the enzyme that inactivates it. These mechanisms are likely to make the mitotic activation of CDC2 (which is accompanied by the nuclear translocation of both CDC2-cyclin-B1 and its activator, CDC25C) and the growth-factor-induced activation of MAP kinase (which, upon sustained activation, concentrates in the nucleus and might thereby partially saturate the relevant MAP-kinase phosphatases) more switch-like. PMID- 9868364 TI - A third fibronectin-type-III domain in the insulin-family receptors. PMID- 9868365 TI - Glycolysis, turbo design and the endocrine pancreatic beta cell. PMID- 9868366 TI - Sequence does specify protein conformation. PMID- 9868367 TI - The HD domain defines a new superfamily of metal-dependent phosphohydrolases. PMID- 9868368 TI - Triple association of CDC25-, Dbl- and Sec7-related domains in mammalian guanine nucleotide-exchange factors. PMID- 9868369 TI - A novel repeat domain that is often associated with RING finger and B-box motifs. PMID- 9868370 TI - Sugar transporters from bacteria, parasites and mammals: structure-activity relationships. AB - Sugar transport across the plasma membrane is one of the most important transport processes. The cloning and expression of cDNAs from a superfamily of related sugar transporters that all adopt a 12-membrane-spanning-domain structure has opened new avenues of investigation, including presteady-state kinetic analysis. Structure-function analyses of mammalian and bacterial sugar transporters, and comparisons of these transporters with those of parasitic trypanosomatids, indicate that different environmental pressures have tailored the evolution of the various members of the sugar-transporter superfamily. Subtle distinctions in the function of these proteins can be related to particular amino acid residue substitutions. PMID- 9868371 TI - Structural organization of MAP-kinase signaling modules by scaffold proteins in yeast and mammals. AB - MAP-kinase signaling pathways are activated by multiple extracellular stimuli. The specificity of activation and function of MAP-kinase signaling modules is determined, in part, by scaffold proteins that create multienzyme complexes. In Saccharomyces cerevisiae, two MAP-kinase-scaffold proteins have been identified. Recent studies of mammalian cells have also led to the identification of putative scaffold proteins. These scaffold proteins appear to facilitate MAP-kinase activation, in response to specific physiological stimuli, and to insulate the bound MAP-kinase module against activation by irrelevant stimuli. Scaffold proteins are therefore critical components of MAP-kinase modules and ensure signaling specificity. PMID- 9868372 TI - Mobile DNA elements: controlling transposition with ATP-dependent molecular switches. AB - Nucleotide-binding proteins are often used as molecular switches to control the assembly or activity of macromolecular machines. Recent work has revealed that such molecular switches also regulate the spread of some mobile DNA elements. Bacteriophage Mu and the bacterial transposon Tn7 each use an ATP-dependent molecular switch to select a new site for insertion and to coordinate the assembly of the transposition machinery at that site. Strong parallels between these ATP-dependent transposition proteins and other well-characterized molecular switches, such as Ras and EF-Tu, have emerged. PMID- 9868373 TI - The origin of life--a review of facts and speculations. AB - Three popular hypotheses attempt to explain the origin of prebiotic molecules: synthesis in a reducing atmosphere, input in meteorites and synthesis on metal sulfides in deep-sea vents. It is not possible to decide which is correct. It is also unclear whether the RNA world was the first biological world or whether some simpler world preceded it. PMID- 9868374 TI - DOMO: a new database of aligned protein domains. PMID- 9868375 TI - The insulin connection: Dorothy Hodgkin and the Beijing Insulin Group. PMID- 9868376 TI - Assassins. PMID- 9868377 TI - Utilizing an epidemiological profile for HIV prevention community planning in West Virginia. AB - An epidemiologic profile revealed that approximately 1,000-1,600 persons in West Virginia are living with HIV, and that it is the 5th leading cause of death among persons ages 25-44. HIV is also the leading cause of death in black males ages 25 44 in WV, and blacks are disproportionately affected by HIV/AIDS (composing about 3.1% of the general population and 17% of the AIDS population). This is most marked in Kanawha, Raleigh, and McDowell counties. The predominant mode of exposure reported from 1984-1993, and more recently from 1991-1993, has been men having sex with men (MSM), accounting for 57% of all AIDS cases. A substantial proportion of all cases (13%) were attributed to injecting drug use (IDU), especially in Public Health District 1 where 63% of women infected with HIV reported IDU as a risk behavior. It is difficult to draw conclusions about populations at risk in WV because of the large proportion of HIV cases reported without risk behaviors (26.5%). However, among men, most cases initially reported without risk are eventually reclassified to the MSM and IDU categories. Among women, most cases first reported without risk are eventually changed to the IDU and heterosexual contact categories. Map analyses by Public Health District and county suggest some clustering of cases in the southern region of the state which may be secondary to racial/ethnic and sociodemographic factors. PMID- 9868378 TI - Schedules for Clinical Assessment in Neuropsychiatry (SCAN): introduction to WV's mental health community. AB - The Schedules for Clinical Assessment in Neuropsychiatry (SCAN) is a semi structured clinical interview used by trained clinicians to assess and diagnose psychiatric disorders among adults. The SCAN core is the Present State Examination (PSE) which has been developed and tested globally during the past four decades with good validity and reliability. The SCAN was developed within the framework of the World Health Organization (WHO) and the National Institute of Mental Health (NIMH) Joint Project on Diagnosis and Classification of Mental Disorders, Alcohol and Related Problems (1). The use of the SCAN gives the flexibility to diagnose mental disorders based on the current International Classification of Disease (ICD), Diagnostic and Statistical Manual (DSM) systems or other diagnostic systems that may develop in the future. A major purpose of the SCAN is to allow comparisons of psychiatric diagnoses to be made across the world. PMID- 9868379 TI - The treatment of breast cancer in the elderly: a community hospital experience. AB - The physician's dilemma in choosing the optimal management for breast cancer in the elderly arises from long-held beliefs that elderly patients 1) have a limited life expectancy from comorbid conditions other than breast cancer and 2) do not tolerate standard treatment. To determine if these beliefs were indeed reflective of the clinical behavior of breast cancer in the elderly, we reviewed the hospital and tumor registry records of 182 women age 70 years and older who received treatment for locoregional invasive breast cancer (Stage I and II) at CAMC between 1977 and 1987. The median time interval between diagnosis and last contact or death was 82 months. Actuarial survival was calculated on the basis of this interval Statistical Analysis System (SAS). Eighty-five percent of patients underwent mastectomy and 15% had breast conservation surgery. There was only one postoperative death. Forty-eight (26%) of 182 patients received systemic adjuvant therapy. The breast cancer-specific survival rate of all patients in this study was 84% at five years and 71% at 10 years. The 5-year and 10-year breast cancer specific survivals were 94% and 86% for stage I patients respectively and for stage II patients the five-year and 10-year survivals were 74% and 56% respectively. Our results indicate that the majority of elderly women with breast cancer can tolerate standard surgical therapy and survive disease-free for many years. PMID- 9868380 TI - [Peptide synthesis aiming at elucidation and creation of protein functions]. AB - The recent development of molecular biology has been elucidating outlines of the cross-talk of biomolecules. The understanding of the function of these biomolecules from the viewpoint of chemistry is now demanded not only for the understanding of biological systems but also for the creation of novel functional molecules. Here two topics are described about peptide synthesis aiming at the elucidation and the creation of protein functions. The first topic is the development of approaches for the synthesis of Tyr (SO3H)-containing peptides. Tyrosine sulfation is one of the most popular protein post-translational modifications. Synthetic peptides are of great help for the elucidation of the biological significance of tyrosine sulfation. We have developed two approaches for the efficient synthesis of tyrosine sulfate [Tyr (SO3H)]-containing peptides. The first approach employs a dimethylformamide-sulfur trioxide (DMF-SO3) complex as a sulfating agent and safety-catch protecting groups for the selective sulfation of tyrosine in the presence of serine. The second approach employs the direct introduction of Tyr(SO3H) into the peptide chain in the form of Fmoc Tyr(SO3Na) followed by deprotection at 4 degrees C in trifluoroacetic acid. These approaches were successfully applied for the synthesis of cholecystokinin (CCK) related peptides. The second topic deals with new approaches for the creation of artificial proteins through assembling alpha-helical peptides via selective disulfide or thioether formation. Approaches to assemble individual peptide segments on a peptide template were also developed. Four peptides corresponding to the transmembrane segments of the sodium channel (S4 in repeat I-IV) were assembled on a peptide template to give a protein having ion channel activity with rectification. PMID- 9868381 TI - [Effect of tableting conditions on the sticking of tablet using ibuprofen]. AB - Influence of tableting conditions on sticking was investigated using two formulations, the one formulation containing ibuprofen (formulation A) and the other being the standard formulation (formulation B). Sticking was observed on formulation A, and was not observed on formulation B. The degree of sticking was decreased with increase in compression pressure, and increased with compression speed. The shape of punch face was influenced the degree of sticking (sugar concave < bebel < flat). It was thought that sticking was observed when the adhesion between tablet and punch was larger than that among tableting granules. We investigated the pressure placed on a scraper (SCR), the adhesion between granules and metal, and boring hardness of tablet. The results showed that sticking was not observed when the SCR and/or the adhesion were small enough or boring hardness of tablet was high enough. Furthermore, it was suggested that boring hardness distribution would influence sticking. PMID- 9868382 TI - [Quantitative determination of constituents in various licorice roots by means of high performance liquid chromatography]. AB - During the course of our studies on the chemical constituents of botanically identified Chinese licorice roots of various origins, we have identified 15 kinds of saponins and 49 kinds of flavonoids (including flavonoid glycosides). With these chemical constituents at hand, we have carried out the chemical evaluation studies of licorice roots available in Japan by means of high performance liquid chromatography (HPLC). In the HPLC analysis using an ODS reversed-phase column, a combination of 1% aqueous acetic acid and acetonitrile was used as the mobile phase in a gradient solvent system, and the eluate was monitored with UV absorption at 254 nm (for saponins with a diene or a conjugated carbonyl moiety, liquiritin, and liquiritin apioside) or at 350 nm (for other flavonoid constituents). By our analytical method described here, quantitative analyses of saponin and flavonoid constituents were effected simultaneously. We have found an interesting piece of information concerning the origin of parent plants and their characteristic constituents such as saponins and flavonoids. PMID- 9868383 TI - [Chemotaxonomy of the Rabdosia in Japan by diterpenoids]. AB - Many diterpenoids containing mostly ent-kaurenoids have been isolated from the Rabdosia (Labiatae). The relation of the diterpenoids with its species in Japan has been studied. The ratio of diterpene types and the values of oxidation state in each plant were calculated. These species were found to be divided into four groups by the cluster analysis of the values of oxidation state of C-1 to C-20 in the diterpenoids of the Rabdosia. PMID- 9868384 TI - Human tetranectin: methodological and clinical studies. AB - From its discovery in 1986 tetranectin (TN) has been suggested to participate in proteolytic processes through its binding to plasminogen, which enhances the activation of plasminogen to plasmin. Because extracellular proteolysis is an important factor in the ability of malignant cells to infiltrate normal tissues and metastasize, TN was considered to be a potential marker for this proteolysis. We have studied the variations in blood and tissue levels of TN in clinical conditions such as cancer and infection, where increased proteolysis can be expected. Five monoclonal antibodies (MAbs) were produced against human TN, and our study is the first report of stable hybridomas producing MAbs against human TN. All the MAbs reacted with epitopes located within aa-residues 50-181 of the primary sequence. In relative epitope mapping with enzyme immuno assay and isotachophoresis the five MAbs defined two independent epitope groups. Different combinations of MAbs were suitable for enzyme immuno assays and two MAbs could be used for immunohistochemical detection of TN in both fresh frozen and paraffin embedded tissues. The MAbs will facilitate future studies on structure, function, clinical significance and immunolocalization of TN. In primary ovarian cancer neither s/p-TN nor CA 125 were found valuable for diagnosis of localized cancer versus benign tumors. However, s/p-TN combined with CA 125, increased both sensitivity and specificity. S/p-TN should therefore be considered one of the screening markers in conjunction with CA 125, or other comparable markers, in future ovarian cancer screening research studies. Preoperative s-TN was significantly correlated to residual tumor and survival in ovarian cancer patients undergoing second or third look surgery. In conjunction with CA 125 and CASA the predictive value of TN for residual tumor was greatly improved, as the markers were found to supplement each other. If the second look operation had been restricted to patients who had a marker negative test, up to 37% would have avoided surgery. We therefore recommend that these tests are included as potential selection parameters in other studies evaluating second-look surgery. Low p-TN concentration and heavy extracellular staining of TN in the tumors was significantly correlated with a poor prognosis for patients with localized stage I or II or advanced primary ovarian cancer. The prognostic information can be especially valuable for patients with a localized ovarian cancer stage I or II, because about 20% of these patients, believed to be radically operated later present with relapse. We found the p-TN level to be especially useful for patients with localized cancer, indicating that adjuvant chemotherapy may be considered if the p-TN level is low. For patients with advanced primary ovarian cancer and low p-TN the survival was significantly reduced compared to patients with a higher p-TN. The p-TN level was significantly negatively correlated to the extracellular (stromal) staining of TN in the tumors. A heavy stromal TN staining was correlated with a shortened survival and was an independent prognostic factor in the Cox analyses. Patients with advanced primary cancer and a low p-TN, possibly in combination with a heavy stromal staining of TN in the tumors, should tentatively be offered palliative treatment or experimental chemotherapy. Furthermore, patients receiving chemotherapy may be monitored by s/p-TN measurements, as a decrease in TN concentration indicated recurrence 3.6 months prior to clinical diagnosis. A decrease in TN concentration during chemotherapy may therefore indicate change of treatment. Serum TN was a very strong independent prognostic factor of poor treatment response and a shortened survival in patients with metastatic breast cancer. A low pre-chemotherapy s-TN value together with clinical signs of poor treatment response may be an ominous combination, which may suggest change of treatment. For patients with Dukes' stage PMID- 9868385 TI - A matter of biochemistry. A short history of scientific discovery. PMID- 9868386 TI - [Organ preservation in ORL oncology: myth or reality. The case for laryngeal preservation]. AB - The conservation of a functional larynx is an important goal in the various attempts to improve the treatment of locally-advanced pharyngo-laryngeal cancers. Several axes have been followed: conservative surgery (subtotal laryngectomy), exclusive irradiation (with conventional or non-conventional fractionation), initial chemotherapy (followed by surgery or irradiation) or concomitant chemotherapy-radiotherapy. The numerous studies, including some major randomised trials, dealing with the issue of larynx preservation are reviewed in the present article. A critical comparison of their results and its integration in a consistent approach are difficult, especially because the different options are aimed at different patients. Total laryngectomy and its psychological and social consequences can be avoided in a significant number of patients, although the long term functional outcome is poorly known and the choice of the optimal treatment modality is largely dependent on the expertise and experience of the medical team. PMID- 9868387 TI - [Influence of locoregional irradiation on local control and survival in breast cancer]. AB - Locoregional control is a crucial step in the achievement of breast cancer cure. In ductal carcinoma in situ, breast irradiation significantly reduces the rates of local recurrence whatever the histological subtypes, as demonstrated by the NSABP-B17 trial (25.8% of local recurrences without radiotherapy vs. 11.4% with radiotherapy). In infiltrating breast carcinomas, complementary breast irradiation has been shown to significantly improve the local control and slightly the overall survival in five randomized trials. Following mastectomy, locoregional irradiation clearly reduces the chest wall and nodal relapse rates, especially in case of lesions more than 5 cm or with nodal involvement and/or large lymphatic or vascular emboli. Two recent randomized trials confirmed the benefit of well-adapted locoregional irradiation in all subgroups, especially in patients with one to three axillary involved nodes. In the Danish trial (including premenopausal high-risk women), radiotherapy reduced locoregional relapses from 32 to 9% (p < 0.001) and increased the 10-year survival rate from 45 to 54% (p < 0.001). In the Canadian trial, locoregional relapse rate decreased from 25 to 13% and the 10-year survival rate increased from 56 to 65%. The meta analysis published in 1995 by the EBCTCG showed only a modest benefit due to locoregional irradiation in breast cancer. However, when small or old trials were excluded due to imperfect methodology or inadequate irradiation techniques, the benefit of modern radiotherapy became much more evident in a population of 7,840 patients. Locoregional irradiation appears to be able to reduce the risk of metastatic evolution occurring after local or nodal relapse and must be integrated in a multidisciplinary strategy. Treatment toxicity (especially toxicity due to irradiation of internal mammary nodes) is of special concern, as anthracycline-based chemotherapy is prescribed more often. The use of a direct field, with at least 60% of the dose delivered by electrons alternating with photons is recommended to protect the heart and lungs. PMID- 9868388 TI - [Local recurrences after conservative treatment of breast cancer: risk factors and influence on survival]. AB - AIMS OF THE STUDY: To determine the risk factors for local and distant failure in node-negative breast cancer treated with breast-conservative surgery and radiotherapy and to determine the relationship between these two events. MATERIAL AND METHODS: We retrospectively selected 908 patients who received conservative surgery and radiotherapy but no chemotherapy between 1980 and 1995, for a node negative breast cancer. Patients were divided in two groups according to the status of the margins of resection. All pathology specimens were reviewed. RESULTS: In case of negative margins, the risk factors for local recurrences picked up by the Cox model were histologic multifocality (P = 0.0076), peritumoral vessel invasion (P = 0.021) and age < or = 40 years (P = 0.024), and in case of involved margins, negative oestrogen receptors (P = 0.0012), histologic multifocality (P = 0.0028), and absence of hormonal therapy (P = 0.017). The 10-year local recurrence rate was 18% in case of negative margins and 29% in case of involved margins, although in the latter case patients received high-dose adjuvant radiotherapy. Accordingly, the 10-year distant failure rates were 16% and 27%, respectively. Many arguments suggest that local and distant failures are closely related. CONCLUSION: Patients with histologic multifocality or positive margins are at high risk of local failure and then of distant failure, and require a more aggressive initial treatment. PMID- 9868389 TI - [Radiation and concomitant chemotherapy after surgery for breast cancer: Arcosein study ]. AB - In breast cancer postoperative radiation therapy improves local control rate and survival. When chemotherapy and radiation are indicated, the sequencing of the two treatments is still controversial. Several studies have suggested that adjuvant radiotherapy could be safely delayed until adjuvant chemotherapy was completed. Other studies, most of them retrospective, pointed out that a delay in the initiation of radiotherapy in order to give chemotherapy first may result in an increased rate of local recurrence. Concomitant administration of the two treatments is an attractive alternative. Pilot studies have demonstrated its feasibility. A randomised phase III trial is presently in progress, comparing sequential treatment with chemotherapy first followed by irradiation and radiation versus concomitant treatment. Four-hundred and five patients have been included in 24 months. Toxicity and compliance to the treatment are reported in this evaluation. PMID- 9868390 TI - [Axillary lymph node surgery in 1998]. AB - The need for a systematic axillary clearance in breast cancer is presently under question. Alternative methods include the omission of node biopsy in very small tumours and lymphadenectomy limited to the sentinel node. This article discusses the current procedures in axillary surgery in 1998, with information concerning the relationships between the tumour characteristics and the probability of nodal involvement, the new surgical techniques aiming at reducing morbidity, with special emphasis on sentinel node biopsy, and the therapeutic protocols presently being used at the Institut Curie. PMID- 9868391 TI - [Prognostic factors of infiltrating bladder tumors]. AB - In France, invasive bladder cancer is the most frequent urologic malignancy after prostate carcinoma. The standard treatment of bladder cancer is radical cystectomy. New therapeutic approaches such as chemoradiation combination for a conservative procedure, neoadjuvant or adjuvant chemotherapy are still in development. In this prospect, a rigorous selection of patients is needed. This selection is based on prognostic criteria which could be divided into four groups: 1) the volume of the tumour including the tumour infiltration depth, the nodal status, the presence or not of hydronephrosis and the residual tumour burden after transuretral resection; 2) the histologic aspects including histologic grading, the presence or not of an epidermoid metaplasia, of in situ carcinoma or of thrombi; 3) the expression of circulating tumour cell biological markers; 4) the biologic characteristics of the tumour such as ploidy, presence of cytogenetic abnormalities, expression of Ki67, expression of oncogenes or tumour suppressor genes, expression of telomerase, expression of tumour antigens or growth factor receptors. This paper reviews the prognostic value of these different parameters. PMID- 9868392 TI - [Conservative treatment of bladder cancers with radiotherapy and concurrent chemotherapy: results and perspectives]. AB - Therapeutic strategies for muscle invasive bladder cancer are currently evolving. Transurethral surgery followed by concurrent chemotherapy and radiotherapy is the best treatment for patients not suitable for radical cystectomy. This conservative approach may be proposed for operable patients with similar survival than after cystectomy. Intermediate evaluation of response is necessary, as response is the more discriminant prognostic factor for local control. Non responders would be assigned for immediate cystectomy. Morbidity and results of salvage cystectomy are poorly evaluated and probably depending from time from initial treatment and dose of radiation. PMID- 9868394 TI - [Is there still a role for radical cystectomy in localized, infiltrating bladder cancers in 1998?]. AB - The role of radical cystectomy as the standard treatment of localised infiltrating bladder malignancy is challenged by the development radiotherapy and chemotherapy combinations. The published studies are difficult to compare because of large differences in the patients selection criteria and in the assessment of local involvement (only clinical in combined treatment, based on pathology and therefore unquestionable in surgical series). The advantages of radical surgery are its precedence and a well-established technique, simplified follow-up procedures and seemingly higher survival rates acquired after 3 years. Conversely, the combination of radiotherapy and chemotherapy allows bladder preservation in about 40% of the patients (at 5 years), but only in 23% without bladder relapses. A complete initial endoscopic resection is the best and only prognostic factor of these results which nevertheless needs a very carefully and endless follows. The comparison of the quality of life achieved by both treatment modalities remains insufficiently documented. PMID- 9868393 TI - [Advances in surgical techniques and results of radical cystectomies for bladder cancer. 106 patients]. AB - AIM OF THE STUDY: Retrospective analysis of result of radical cystectomy at Cochin Hospital. PATIENTS AND METHODS: We report the results of a 106 patients series treated by radical cystectomy for bladder carcinoma after a 5-year period follow-up. RESULTS: The extent of the tumour invasion according to pathological analysis was: pT1 or less: 26%, pT2 and pT3a: 33%, pT3b and over: 41%. Morbidity rate was 19% with a 7.5% reintervention rate. Long term complication rate was 31%, concerning essentially ureteral stenosis. A local recurrence or distant metastasis occurred in 35% of patients. Local recurrence rate was 10.7%. Cancer specific survival rates were 88%, 81% and 42% for pT1, pT2-pT3a, and pT3b patients respectively. CONCLUSION: The present results confirm that radical cystectomy is the most effective curative treatment for invasive bladder carcinoma. PMID- 9868395 TI - [Chemotherapy for urothelial infiltrating tumors]. AB - Since 1985, the gold standard regimen for metastatic tumours of the urothelial tract is a combination of methotrexate, vinblastine, doxorubicin and cisplatin (M VAC protocol). Randomised studies have shown the superiority of the M-VAC regimen over cisplatin alone or in combination with cyclophosphamide and doxorubicin. The limited gain in survival shows that, although chemosensitive, metastatic tumours cannot be cured by the available drug combinations. Amongst new cytotoxic drugs in development, paclitaxel and gemcitabine are likely to be involved in the standard regimen of the future. The studies published to date do not support the routine use of chemotherapy in the neo-adjuvant or adjuvant setting for the management of early stage urothelial tumours. PMID- 9868396 TI - [Tumor and individual radiosensitivity. An introduction]. AB - This text is intended to introduce the following articles, which correspond to the lectures given at the "Radiosensitivity" session of the 1998 SFRO Meeting. We first underline the somewhat disappointing results obtained so far for evaluating tumoral radiosensitivity. However, a few new tests could bring some hope in a not too distant future. We then focus on individual radiosensitivity. We first list the syndromes or diseases associated with a known hyperradiosensitivity. We then concentrate on the radiosensitivity tests that are available in 1998, some of them being already used in a few centers in specific situations. However, the "optimal" test is still to be identified. After a brief chapter on the possible modulations of radiosensitivity, we conclude with a few recommendations to the clinician. PMID- 9868397 TI - [Tumor radiosensitivity tests: state of assays in 1998]. AB - Various assays measuring tumor radiosensitivity were developed in the early eighties. These assays were based on cell cultures derived from tumor biopsies. In most studies there was no correlation between measured radiosensitivity and treatment outcome. New assays using new techniques (such as measurement of radiation-induced apoptosis, or fluorescence in situ hybridization) have been developed recently. Preliminary results are encouraging, but correlation with treatment outcome is expected to be difficult to demonstrate as multivarious clinical and biological parameters are involved in loco-regional control. PMID- 9868398 TI - [Individual radiosensitivity and DNA repair proficiency: the value of the comet assay]. AB - Some rare hereditary syndromes demonstrate high cancer risk and hypersensitivity in response to exposures to agents such as ultraviolet or ionising radiation, and are characterised by a defective processing of DNA damage. They highlight the importance of the individual capacity of restoring the genomic integrity in the individual risk associated to exposures. The comet assay, a simple technique that detects DNA strand breaks, requires few cells and allows examination of DNA repair capacities in established cell lines, in blood samples or biopsies. The assay has been validated on cellular systems with known repair defects such as xeroderma pigmentosum defective in nucleotide excision repair, on mutant rodent cell lines defective in DNA single strand break rejoining (XRCC1) (alkaline version) or DNA double strand breaks rejoining (XRCC5/Ku80 and XRCC7/DNAPKcs) (neutral conditions). This assay does not allow to distinguish a defective phenotype in ataxia telangiectasia cells. It shows in homozygous mouse embryo fibroblasts Brca2-/- an impaired DNA double strand break rejoining. Simplicity, rapidity and sensitivity of the alkaline comet assay allow to examine the response of lymphocytes. It has been applied to the analysis of the role of DNA repair in the pathogenesis of collagen diseases, and the involvement of individual DNA repair proficiency in the thyroid tumorigenesis induced in some patients after therapeutic irradiation at childhood has been questioned. Preliminary results of these studies suggest that this type of approach could help for adapting treatment modalities and surveillance in subgroups of patients defective in DNA repair process. It could also have some incidence in the radioprotection field. PMID- 9868399 TI - [Radiation-induced cancers]. AB - The induction of malignant diseases is one of the most concerning late effects of ionising radiation. A large amount of information has been collected form atomic bomb survivors, patients after therapeutic irradiation, occupational follow-up and accidentally exposed populations. Major uncertainties persist in the (very) low dose range i.e., population and workers radioprotection. A review of the biological mechanisms leading to cancer strongly suggests that the vast majority of radiation-induced malignancies arise as a consequence of recessive mutations of tumour-suppressor genes. These mutations can be unveiled by ageing, this process being possibly furthered by constitutional or acquired genomic instability. The individual risk is likely to be very low, probably because of the usual dose level. However, the magnitude of medical exposure and the reliance of our societies on nuclear industry are so high that irreproachable decision making processes and standards for practice are inescapable. PMID- 9868401 TI - [Non-small-cell bronchial cancer: update in 1998]. PMID- 9868400 TI - [Radiotherapy and skin tumors]. AB - Radiotherapy plays an important role in the treatment of skin tumours. For skin carcinomas, external irradiation (kilovoltage X-rays or electrons according to clinical characteristics) is more valuable than interstitial brachytherapy, which is recommended for tumours of the lip and of the nasal vestibule. In mycosis fungoides, total cutaneous electron beam radiation therapy is efficient for patients with limited superficial plaques. In the classical form of Kaposi's sarcoma, radiotherapy can achieve local control whereas it obtains good palliative results in the epidemic form. PMID- 9868402 TI - [Value and limits of surgery in stage IIIB non-small-cell bronchial cancers]. AB - According to the TNM staging system for lung cancers, stage III is divided into IIIA and IIIB. This division was based upon the principle that patients with IIIA disease could theoretically benefit from complete resection, contrasting with IIIB patients for whom surgery is not feasible. The poor prognosis of stage IIIB is largely due to its classical inoperability. From the surgical point of view, stage IIIB can be subdivided into four subgroups: 1) N3 where resection is possible in selected patients through median sternotomy; 2) T4 where extended surgery can be considered in selected patients; 3) N3 + T4; 4) malignant pleural or pericardial effusion contraindicating any radical surgery. Criteria for resectability could be defined to include some IIIB patients in multimodality protocols in which surgery would become possible after induction therapy: definitive inoperability excludes any possibility of surgery, even in cases in which radiotherapy alone or combined with chemotherapy leads to complete remission; immediate inoperability allows patients to be included in protocols evaluating induction treatments designed to render tumours resectable. PMID- 9868403 TI - [Which fractionation and radiation dose for non-small-cell bronchial cancers?]. AB - The optimal radiation schedule (dose, fractionation, volume) must be adapted to the goal of the treatment: cure with radiation alone or a radiochemotherapy approach or palliation. The former implies to deliver a dose of 60 Gy or more in 6 weeks or an equivalent biological doses. This approach should take into account the different prognostic factors (tumor size, extension...) and the technical possibilities (volume of normal tissue irradiated). Doses above 70 Gy should still be reserved for clinical investigation (conformal radiotherapy). PMID- 9868404 TI - [Radiotherapy for non-small-cell bronchial cancers: definitions of volumes, patient selection. Recommendations of the International Association for the Study of Lung Cancer (IASLC)]. AB - Chemoradiation is the standard treatment of unresectable, locally advanced non small cell lung cancer, with a mean dose of 60-66 Gy, excluding escalation dose schemes. The standard treated volume includes primary tumor, ipsilateral hilar and mediastinal nodes, supraclavicular and contralateral nodes as well, regardless of the node status. This work tries to answer the question of the optimal volume to be treated. Drainage routes analysis is in favor of large volumes, while toxicity analysis favors small volumes. Combined modality treatment may increase the observed toxicity. The optimal volume definition is difficult, and requires available conformal therapy tools. Patients selection is another important issue. A volume definition is then attempted, based on the IASLC (International Association for the Study of Lung Cancer) Annecy workshop experience, highlighting the interobservers discrepancies, and suggests basic recommendations to harmonize volume definition. PMID- 9868405 TI - [Radiochemotherapy and radiosensitization for locally advanced non-small-cell bronchial cancer]. AB - Combined chemotherapy and radiotherapy can improve the survival of patients with locally advanced non-small cell lung cancer, when compared to irradiation alone. This survival benefit is essentially due to an increased control of distant micrometastases, whereas local control remains poor. In order to improve local control, new radiotherapy modalities such as 3D conformal treatment, hyperfractionation or accelerated hyperfractionation, are under development. Cytotoxic drugs given at low doses concomitantly to radiotherapy may act as radiosensitizers on the primary tumor. Concomitant chemotherapy at cytotoxic doses and radiotherapy would also allow better control on micrometastases and better local control due to radiosensitization by chemotherapy. However, the concomitant use of chemotherapy and radiotherapy is limited by increased toxicity on normal tissues, more particularly on the esophagus. Randomized comparisons of these modalities versus induction chemotherapy followed by radiotherapy are needed to determine the optimal treatment sequence. PMID- 9868406 TI - [Control of smoking to reduce the incidence of bronchial cancer: application of Evin's law in France]. AB - In France, 30% of adults smoke, and males smoke a little bit more than females. Young people smoke more and more, and 50% of 18-24 years-old males and females smoke. Tobacco causes 60,000 deaths each year in France. Publicity is now forbidden (Evins's law). Price of cigarette has been multiplied by two. Interdiction of smoking in public and closed places is however insufficiently respected. Budget devoted to information, education, and prevention, is also dramatically insufficient. By contrast, due to vigilance of the Comite National Contre le Tabagisme, tobacco sales decreased from 1991 to 1997 of 11.1%, and cigarette sales of 14.5%. French situation must be replaced in European and international context. European regulation on smoking interdiction increases the impact of the French legislation. PMID- 9868407 TI - [Last ICRU recommendations for the prescription, recording and reporting of external bean therapy]. AB - Due to the evolution of radiotherapy techniques and equipment, in 1993 the ICRU decided to modify the guidelines that had been published in the ICRU report 29 (1978) in regard to photon beam therapy and to insert them in Report number 50 (1993). Changes that were introduced mainly concerned the prescription, recording and reporting of external beam therapy with photons. While awaiting for the upcoming publication of the report focusing on external beam therapy with electrons, the ICRU decided to publish a supplement to Report 50, as new insights had been gained after discussions with the Nordic Association of Clinical Physicists (NACP). This supplement includes the following: more accurate definitions of the margins allowing the planned target volume to be derived from the clinical target volume, the internal target volume, the expected volume of organs at-risk and the conformity index. All three points will be included in the report focusing on external beam therapy with electrons as well as extensive discussions relating to the implementation of these new recommendations. PMID- 9868408 TI - [Practical determination of volume and doses in conformal radiotherapy]. AB - The determination of the target volumes and the dose to be delivered are critical medical steps in order to locally control the disease and to limit the risks of normal tissues injury. While many clinical situations are compatible with standardised technique and beam arrangements, the tools provided by modern medical diagnostic imaging procedures and tridimensional dosimetry allow an improved conformation of the irradiated volume and the administered dose to the clinical target volume. However, this approach requires an increased precision in tumour and normal tissues delineation, based on patient immobilisation devices, standardisation of mobile organs repletion status and image acquisition procedures, explicit definition of the biological and physical safety margins, definition of acceptable levels of dose heterogeneity in the planning target volume and appropriate quality assurance procedures. PMID- 9868409 TI - [3D automatic expansion: clinical application]. AB - The determination of the various volumes (GTV: gross target volume, CTV: clinical target volume, PTV: planned target volume) recommended by the ICRU 50 report is a critical step in conformal treatment planning, since treatment optimisation procedures and documentation rely on accurate dose-volume histograms. The shape and the size of the CTV vary with the computer algorithm, the patient image acquisition parameters, the definition of the GTV and the margins surrounding it. The automatic expansion programs included in commercially available treatment planning system require careful validation and control before and during their routine use by the clinicians. Significant differences have been observed between 2D- and 3D-based expansions, with a usual underestimation of the PTV by 2D algorithms. PMID- 9868410 TI - Prevalence of coronary blood flow reserve abnormalities among patients with nonobstructive coronary artery disease and chest pain. AB - OBJECTIVE: To determine the prevalence of abnormalities in endothelium-dependent and endothelium-independent coronary flow reserve among patients with non obstructive coronary artery disease and chest pain. MATERIAL AND METHODS: We studied endothelium-dependent (after infusion of 10(-6) M to 10(-4) M of acetylcholine) and endothelium-independent (after administration of 18 to 36 micrograms of adenosine) coronary flow reserve among patients with nonobstructive coronary artery disease and chest pain who were undergoing assessment at Mayo Clinic Rochester. Coronary blood flow was derived from coronary artery diameter assessed by quantitative angiography and Doppler flow velocities. RESULTS: The cohort consisted of 203 patients (158 female and 45 male patients), who ranged in age from 17 to 78 years (mean, 51). Most patients (92%) had at least one risk factor for atherosclerosis; a substantial proportion had undergone extensive cardiac and noncardiac evaluation. Whereas 41.5% of the patients had normal coronary flow reserve, 58.5% had an abnormal response: 11.3% an impaired response to adenosine (flow velocity ratio of 2.5 or less), 29.2% an impaired response to acetylcholine (flow reserve ratio of 1.5 or less), and 18% a combined abnormality. No correlation (r2 = 0.03) was noted between endothelium-dependent and endothelium-independent flow reserve. CONCLUSION: Most study patients with chest pain and nonobstructive coronary artery disease undergoing coronary vasomotor evaluation had risk factors for coronary artery disease and diverse abnormalities in endothelium-dependent or endothelium-independent coronary flow reserve (or both). These findings underscore the need for a comprehensive assessment. PMID- 9868412 TI - Bone mass and muscle strength in female college athletes (runners and swimmers). AB - OBJECTIVE: To determine whether female college athletes had increased muscle strength and bone mass in comparison with age-matched nonathletic female subjects and, if so, whether participation in weight-bearing versus non-weight-bearing exercise made a difference. MATERIAL AND METHODS: We performed a comparative statistical analysis of the bone mineral density (BMD) of the total body, lumbar spine, and femoral neck, maximal oxygen uptake (VO2max), muscle strength, and level of physical activity in 21 runners, 22 swimmers, and 20 control subjects. The study participants were female college students, 18 to 24 years old, who had had more than 8 normal menstrual cycles during the past year. RESULTS: Statistical analyses showed significantly higher VO2max in the two athletic study groups than in the control subjects (P < 0.0001). No significant difference in BMD was noted among the three groups. Total body BMD (r = 0.30; P = 0.02) and femoral neck BMD (r = 0.39; P = 0.002) were positively correlated with weight bearing activity but not with non-weight-bearing activity. VO2Max (an index of physical fitness) was positively correlated with femoral neck BMD (r = 0.33; P = 0.009) and trochanteric BMD (r = 0.29; P = 0.021). Shoulder muscle strength (determined by isokinetic dynamometry) was positively correlated with total body BMD (r = 0.34; P = 0.007) and lumbar spine BMD (r = 0.28; P = 0.028). Swimmers had higher muscle strength in the back and upper extremities than did runners and control subjects. Hip girdle muscle strength was not significantly different among the three groups. Total body BMD had a positive correlation with percentage of body fat and height. Lumbar spine BMD was higher in subjects who had previously used oral contraceptives. The athletes had a lower percentage of body fat, were less likely to have used oral contraceptives, and had fewer years of normal menses than did the control subjects. CONCLUSION: Our study shows that (1) total body BMD and femoral neck BMD were significantly higher in the study group that performed weight-bearing exercises than in control subjects, (2) swimming exercise had no effect on BMD, and (3) although swimming is not a bone-building exercise, it can significantly improve shoulder, back, and grip muscle strength. PMID- 9868411 TI - Clinical and laboratory indices that enhance the diagnosis of postural tachycardia syndrome. AB - OBJECTIVE: To identify clinical and laboratory indices that improve the diagnosis of the postural tachycardia syndrome (POTS). DESIGN: We assessed associations of orthostatic intolerance by using multivariate regression analysis. MATERIAL AND METHODS: We evaluated autonomic symptoms and autonomic function in 30 patients with POTS, 30 patients with mild orthostatic intolerance, and 19 age- and gender matched control subjects. Indices of parasympathetic and sympathetic functions were analyzed on the basis of (1) autonomic function tests (head-up tilt), (2) oscillations at respiratory and nonrespiratory frequencies (0.01 to 0.09 Hz) in R R interval and blood pressure (Wigner distribution), and (3) deterministic component (rescaled range analysis). RESULTS: The four clinical and laboratory indices that independently supported the diagnosis of POTS are as follows: (1) orthostatic heart rate during the first minute of head-up tilt, (2) autonomic deficit (adrenergic autonomic score), (3) loss of spectral powers in R-R interval during head-up tilt at the fifth minute, and (4) severity of orthostatic dizziness, fatigue, palpitations, and shortness of breath. CONCLUSION: Enhancing the sensitivity and specificity of the diagnosis of POTS should be possible by using these four indices. A hyperadrenergic state and distal neuropathy, affecting adrenergic sympathetic cardiovagal fibers, seem to be involved in the pathophysiology of POTS. Certain features suggest brain-stem dysregulation. PMID- 9868413 TI - Radioimmunoassay for glutamic acid decarboxylase (GAD65) autoantibodies as a diagnostic aid for stiff-man syndrome and a correlate of susceptibility to type 1 diabetes mellitus. AB - OBJECTIVE: To establish and validate a double-antibody radioimmunoassay (RIA) for detecting serum auto-antibodies against glutamic acid decarboxylase (GAD65). This enzyme catalyzes synthesis of the neurotransmitter gamma-aminobutyric acid in neurons and pancreatic islet cells. MATERIAL AND METHODS: We compared the frequency of GAD65 and other "thyrogastric" autoantibodies in adult patients with stiff-man (Moersch-Woltman) syndrome, type 1 diabetes, or polyendocrine disorders and in healthy subjects. The frequency of pancreatic islet cell antibody (ICA) detection was also assessed. The GAD65 RIA was validated by testing blinded samples, by confirming the specificity of low-titered positive results by "cold" antigen inhibition, and by comparing the RIA results with results of a kit assay incorporating staphylococcal protein A as immunoprecipitant. Recombinant GAD65 protein labeled with 125I was used as antigen, and a combination of anti-human IgG and IgM was used as immunoprecipitant. Seropositivity was determined for ICA and gastric parietal cell antibodies by indirect immunofluorescence assays and for thyroid peroxidase (microsome) and thyroglobulin antibodies by agglutination assays. RESULTS: We detected GAD65-specific antibodies in all but 1 of 46 local patients with stiff-man syndrome (98%); 16 had evidence of diabetes. Positive values exceeded 20 nmol/L in 96%, and 89% were ICA-positive; 76% had additional thyrogastric antibodies. Of 41 patients with type 1 diabetes (17 local and 24 workshop serum specimens), 33 were GAD65 antibody-positive (80%); 85% of these positive values were 20 nmol/L or lower. Only 18% of sera from patients with type 1 diabetes were ICA-positive, but 59% had other thyrogastric autoantibodies. Of 20 patients with autoimmune endocrinopathies without diabetes or stiff-man syndrome, 35% were GAD65 antibody-positive, 5% were ICA-positive, and 90% were thyrogastric antibody-positive. Of 117 healthy control subjects, 8% were GAD65 antibody-positive, and a third of those had other thyrogastric antibodies (14% overall); none was ICA-positive. CONCLUSION: Seropositivity in the double antibody RIA for GAD65 autoantibody is a sensitive and specific marker of predisposition to type 1 diabetes and related organ-specific autoimmune disorders. As such, this RIA is complemented by assays for thyroid and gastric parietal cell autoantibodies. PMID- 9868415 TI - Reconstructing the human face by using the tissue-integrated prosthesis. AB - Reconstructing the human face with autogenous tissue is extremely difficult. A prosthetic device is often used, but retention is a problem. The tissue integrated prosthesis has resolved this issue, however, and the technique is currently available worldwide. The tissue-integrated prosthesis--that is, a prosthesis supported (retained) by osseointegrated implants in the underlying bone--originated during the 1950s, when Branemark began his laboratory studies of tissue healing in Sweden. The term "osseointegration" was introduced in 1977 and later was defined as "direct anchorage of an implant by the formation of bony tissue around the implant without the growth of fibrous tissue at the bone implant interface." Early studies during the 1980s and 1990s documented the success of reconstruction of the human face with use of the tissue-integrated prosthesis. Herein we describe three cases in which this reconstructive procedure was successful. PMID- 9868414 TI - Tactile illusion-produced number perception in blind and sighted persons. AB - OBJECTIVE: To determine whether tactile receptive communication is possible in both sighted and blind persons through the use of a mechanical device that creates the perception of continuous contact through sequential points of vibration on the skin and whether the ability of these two groups differs. MATERIAL AND METHODS: The cohort consisted of 20 sighted and 20 blind subjects who had no prior exposure to a tactile communication device that produces a tactile illusion. Each subject was presented number sequences created by the tactile illusion in the form of single-digit, three-digit, and four-digit sequences, and the number of trials needed for correct identification was recorded. RESULTS: On average, the sighted subjects tended to identify the digit sequences sooner than did the blind subjects by almost one trial; however, this difference was not statistically significant. Younger subjects and female subjects also tended to identify the digit sequences sooner. Nevertheless, the overall correlation between age and number of trials across all replications was relatively minor. CONCLUSION: Tactile receptive communication is possible through the use of a mechanical device used to create tactile illusions. The tactile illusion of numbers is equally perceived by blind and sighted persons. PMID- 9868416 TI - Jonas E. Salk--discoverer of a vaccine against poliomyelitis. PMID- 9868417 TI - The Philadelphia chromosome negative chronic myeloproliferative disorders: a practical overview. AB - The chronic myeloid disorders are collectively characterized by a stem cell origin of the clonal process and a variable tendency to undergo indigenous disease transformation and leukemic conversion. Classification of the chronic myeloid processes is based primarily on the presence or absence of the Philadelphia chromosome (bcr-abl translocation) and secondarily on the morphologic picture of the bone marrow in conjunction with the clinical manifestation. Essential thrombocythemia (ET), polycythemia vera (PV), and agnogenic myeloid metaplasia (AMM) constitute the classical group of bcr-abl negative chronic myeloproliferative disorders. PV is characterized by a clonal increase in red blood cell mass, AMM by bone marrow fibrosis, and ET by thrombocytosis. Most of these features, however, are not diagnostically specific, and secondary causes of erythrocytosis, thrombocytosis, and bone marrow fibrosis must be excluded. Treatment may be deferred or limited to phlebotomy alone in some patients with ET or PV, respectively. In contrast, thrombosisprone patients with PV or ET require drug therapy, and new platelet-lowering agents are increasingly being used. In this article, current diagnostic and therapeutic issues of ET, PV, and AMM are discussed. PMID- 9868418 TI - Benign superior vena cava syndrome due to suppurative mediastinal lymphadenitis: anterior mediastinoscopic management. AB - The benign superior vena cava syndrome is an uncommon medical emergency. We describe a case of the superior vena cava syndrome caused by suppurative mediastinal lymphadenitis. The organisms isolated from various cultures were group C beta-hemolytic Streptococcus, Fusobacterium species, Corynebacterium species, Eikenella corrodens, and Streptococcus milleri. These anaerobic bacteria are part of the normal flora of the upper respiratory tract and the oral cavity. Anterior mediastinoscopy through the right parasternal approach was used to drain the anterior mediastinal abscess and to establish the etiologic factor. PMID- 9868419 TI - Hepatic lymphangiomatosis mimicking polycystic liver disease. AB - Hepatic lymphangiomatosis is a rare disorder characterized by cystic dilatation of the lymphatic vessels in the hepatic parenchyma. It can occur in the liver alone, in the liver and spleen, or in multiple organs. Clinically, diagnosis can be difficult because of the rarity and protean manifestations of this disorder. We describe a 53-year-old woman with hepatic lymphangiomatosis in whom polycystic liver disease had been previously diagnosed. In addition, we review 12 cases of hepatic, splenic, and hepatosplenic lymphangiomatosis with or without systemic lymphangiomatosis and discuss the differential diagnosis. PMID- 9868420 TI - Acute disseminated encephalomyelitis after accidental injection of a hog vaccine: successful treatment with plasmapheresis. AB - Acute disseminated encephalomyelitis, an inflammatory demyelinating disease of the central nervous system, can occur after viral infections or vaccinations. We report the clinical and neuroimaging findings in a 52-year-old man in whom acute disseminated encephalomyelitis developed after accidental self-injection of an industrial hog vaccine. The protracted and progressive clinical course, despite high-dose parenteral corticosteroid therapy, was altered by aggressive plasmapheresis. PMID- 9868421 TI - Medically induced gingival hyperplasia. AB - Gingival hyperplasia or gingival overgrowth is a common occurrence in patients taking phenytoin, cyclosporine, or calcium channel blockers. Speech, mastication, tooth eruption, and aesthetics may be altered. Controlling the inflammatory component through an appropriate oral hygiene program may benefit the patient by limiting the severity of the gingival overgrowth. In patients in whom gingival overgrowth is present or may be anticipated, recognition of this condition and referral to a general dentist or periodontist are appropriate steps to management. The physician's awareness of the potential for development of overgrowth and the dental practitioner's role in attempting to prevent or minimize this problem are important aspects. In this article, we discuss the medications associated with gingival hyperplasia and describe appropriate recommendations. PMID- 9868422 TI - 69-year-old man with fever, diarrhea, and abdominal pain. PMID- 9868423 TI - Antifungal agents. Part I. Amphotericin B preparations and flucytosine. AB - Traditionally, amphotericin B has been the cornerstone of antifungal treatment. Toxicity, however, is a major dose-limiting factor of amphotericin B deoxycholate. Nevertheless, it continues to have a major role in the treatment of deep-seated mycotic infections. Recently, less nephrotic lipid formulations, including amphotericin B lipid complex, amphotericin B cholesteryl sulfate, and liposomal amphotericin B, have been introduced. The pharmacologic properties, main indications, recommended dosages, related costs, and adverse effects of these various preparations are summarized in this review. Orally administered flucytosine is useful in certain infections, particularly cryptococcal meningitis, but it should be used with caution in patients with renal insufficiency. PMID- 9868424 TI - Can measures of coronary dynamics explain chest pain without coronary artery disease? PMID- 9868488 TI - Hereditary palmoplantar keratoderma and dermatophytosis in the northernmost county of Sweden (Norrbotten). AB - Clinical reports of hereditary palmoplantar keratoderma are generally based on a limited number of patients. In 1967 the prevalence in the northernmost county of Sweden (Norrbotten) was shown to be 0.55%. In 1982 it was possible to trace half of the original propositi from that study. Among these families, a severe clinical form with a presumed recessive inheritance could be distinguished. The clinical pictures in relatives of the original propositi were described, and other diseases were listed together with those in patients from previously performed studies. The frequency of dermatophytosis was 36.2%, which was equal to a prevalence of 37.6%. T. mentagrophytes occurred significantly more often and immunological factors, such as increased presence of blood group A, specific dermatophyte IgG antibodies, precipitating antibodies and an immunological in vitro reaction to keratin, supported differences in the distribution of dermatophytes. However, the amount of keratin was considered the most important factor for the affinity of dermatophytes to the palms and soles. A vesicular eruption along the hyperkeratotic border and a mononuclear cell infiltrate were often reported. Such reactions were interpreted as immunological reactions to dermatophytosis. Scaling and fissuring were considered clinical signs of dermatophyte infections and not a part of the originally reported clinical picture. Results of the histopathological study corresponded to previously reported descriptions of the Unna-Thost variety. However, it has recently been reported that the histopathological picture of this variety was based on histopathological features of epidermolytic palmoplantar keratoderma. The existence on the Continent of the Unna-Thost variety was therefore questioned. Histopathological features of epidermolytic palmoplantar keratoderma were not found in the County of Norrbotten and the designation "Diffuse HPPK type Norrbotten" has therefore been proposed. The histopathological picture of the presumed recessive variety did not differ from that of the dominant variety but ultrastructural characteristics differentiated it from Mal de Meleda and the dominant variety. It was therefore concluded that a new variety with a presumed recessive inheritance was found. PMID- 9868489 TI - An epidemiological study of hand eczema. I. Prevalence and cumulative prevalence among hairdressers compared with a control group of teachers. AB - The aim of this cross-sectional study was to estimate the prevalence of dermatitis of the hands and/or forearms among hairdressers compared with a control group of elementary school teachers. Questionnaires were sent to 818 hairdressers and 816 teachers. The response rates were 83% and 65%, respectively. The mean age for those completing the questionnaire, was 28 years (range: 17-71) for the hairdressers and 45 years (range: 20-70) for the teachers. The mean number of months employed in these occupations was 105 (range: 1-672) and 186 (range: 0-516), respectively. Forty-two per cent of the hairdressers and 23% of the teachers suffered or had been suffering from exanthema of the hands and/or forearms. Sixty-one and 15%, respectively, related the skin disease to work. PMID- 9868490 TI - An epidemiological study of hand eczema. II. Prevalence of atopic diathesis in hairdressers, compared with a control group of teachers. AB - The aim of this cross-sectional study was to estimate the prevalence of atopic dermatitis in hairdressers compared with a control group of elementary school teachers. Both groups consisted of employees who reported exanthema on the hands and/or forearms at the time of completing a questionnaire. The mean age for the affected hairdressers was 26 years (range: 17-63) and for the teachers 44 years (range: 28-65). The mean duration of employment was 85 months for the hairdressers (range: 2-552) and 177 months for the teachers (range: 24-360). The frequency of atopic dermatitis was 12.7% (C.I.: 6.3-19.2) in the group of hairdressers and 25% (C.I.: 7.7-42.3) in the group of teachers. No difference was observed between the groups regarding atopic mucosal symptoms or familial atopy. PMID- 9868491 TI - An epidemiological study of hand eczema. III. Characterization of hairdressers with and without hand eczema, regarding demographic factors and medical histories. AB - The aim of this study was to compare, by stratification, subgroups of hairdressers. Three groups were considered: Group I (102 persons) consisted of hairdressers reporting skin affection at the time of completing an earlier mailed questionnaire (point prevalence). Group II reported exanthema prior to completing, while group III reported not having had exanthema at all (51 and 50 persons, respectively). No significant difference was found between these groups with regard to age and duration in work. The mean number of months since start of disease was significantly higher for group I (64 (range: 0.5-552)) than group II (7 (range: 0.6-38)). The frequency of atopic dermatitis was 12.7% (C.I: 6.3-19.2) in group I, compared with 3.9% (C.I: 0.0-9.2) in group II and none in group III. This difference between groups was significant. A significantly higher extent of sick leave was found in group I, compared to group II. PMID- 9868492 TI - An epidemiological study of hand eczema. IV. Degree and pattern of eczema in affected hairdressers, with and without atopic symptoms, compared with a control group of affected teachers. AB - The aim of this study was to show degree and pattern of hand eczema in affected hairdressers (n = 69), compared with a control group of teachers (n = 19). Both groups consisted of employees having eczema, confirmed by the visit of a doctor at the working place. The mean age for the affected hairdressers was 27 years (range: 17-63) and for the teachers 43 years (range: 27-57). The mean duration of employment was 96 months for the hairdressers (range: 2-552) and 182 months for the teachers (range: 25-336). The localization of eczematous lesions, presence of efflorescenses and extent of skin affection were recorded by constructing scores for these variables. The mean number of localizations was significantly higher in hairdressers than in teachers; this was caused by greater affection of fingers in hairdressers compared to teachers. Seventy-five percent of the hairdressers and 79% of the teachers had only modest or very moderate erythema. Fourteen percent and 5%, respectively, had vesicular dermatitis. The right upper limb was most affected in both hairdressers and teachers. Three subgroups of hairdressers were considered: hairdressers with atopic dermatitis, those with only atopic mucosal symptoms and those without atopic symptoms. No significant differences were found between these groups, except a significant higher finger involvement in atopics compared to non-atopics. PMID- 9868493 TI - An epidemiological study of hand eczema. V. Prevalence among hairdresser trainees, compared with a general population of hairdressers. AB - The aim of this study was to show degree and pattern of hand eczema in affected hairdresser trainees (n = 52), compared with a general population of affected hairdressers (n = 69). The mean ages were 18 years (range: 16-25) for the trainees and 27 years (range: 17-63) for the hairdressers. For the latter group, the mean number of months spent in profession was 96 (range: 2-552) and the number of working hours per week was 36 (range: 4-50). Twenty-seven per cent of all trainees (67/246) were reported to have had initial of eczematous lesions on the hands and/or forearms during their apprenticeship. The extent of skin affection, as assessed by scores for localization and distribution of eczema, was significantly lower in trainees, compared with the general population of hairdressers. Patch test revealed sensitization to nickel to be prominent in both groups; 34% among affected hairdressers and 26% among affected and non-affected trainees. Of the hairdressers, 3% had allergy to hair dye (2.5-toluenediamine), while 8% were sensitized to glyceryl monothioglycolate. None of the trainees were sensitized to those two chemicals. PMID- 9868494 TI - An epidemiological study of hand eczema. VI. A follow-up of hairdresser trainees, with the focus on various health complaints. AB - The aim of this follow up study was to show how career dropouts among young hairdressers are related to various health complaints. Questionnaires were sent to 286 hairdressers, who had been registered as trainees three to five years earlier. The forms were returned from 124 hairdressers, of whom 26 (21%) reported that they stopped working as hairdressers because of various health complaints. Eight (6.5%) left their job because of skin conditions. PMID- 9868507 TI - Moroxydine: the story of a mislaid antiviral. AB - A review is presented which began as a literature survey listing verbatim over fifty abstracts and references to the use of moroxydine in the treatment of a number of viral disorders throughout Europe and Scandinavia between 1960 and 1985. The list was first circulated in 1990 and was revised six times as new information became available. The sources are unusual in their diversity and originally appeared in several other languages besides English. Moroxydine may be effective against a number of DNA and RNA viruses, influenza being the original application of the substance when it emerged in the late 1950's. The drug received some attention around 1960, and the introduction of thalidomide at around the same time (1961) is one likely explanation for its current obscurity. Side-effects are reported to be mild and infrequent, and evidence exists that the substance possesses other characteristics which plead for further investigation. PMID- 9868506 TI - Allergenic components in modified and unmodified rosin. Chemical characterization and studies of allergenic activity. AB - Gafvert, E. 1994. Allergenic components in modified and unmodified rosin. Chemical characterization and studies of allergenic activity. Acta Dermato Venereologica. Suppl. 184. 36pp. Uppsala. Unmodified rosin (colophony) is a well known cause of contact allergy (delayed type hypersensitivity). Rosin is obtained from coniferous trees and consists mainly of diterpenoid resin acids. Most rosin used in technical products is chemically modified. In the common modification of rosin with maleic anhydride, the major product formed is maleopimaric acid (MPA). MPA was identified in experimental sensitization studies as a potent contact allergen. MPA is also formed when rosin is modified with fumaric acid at high temperature and with prolonged heating. The amounts of MPA in technical quality rosins modified with maleic anhydride or fumaric acid might be enough to sensitize individuals handling these rosins. The major product of the modification of rosin with fumaric acid, fumaropimaric acid (FPA), did not elicit any reactions in the animals tested. In another common rosin modification, glycerol esterification, the major product formed was identified as glyceryl triabietate (GTA). In an experimental sensitization study none of the animals reacted to GTA. However, a minor product formed, glyceryl 1-monoabietate (GMA) showed sensitizing capacity. The presence of new contact allergens due to the modification, together with remaining unmodified material, contributes to the risk of developing allergy from contact with these types of rosin. A new main contact allergen in unmodified rosin was identified; 13,14(beta)-epoxyabietic acid. The allergenicity of this epoxide was comparable to that of an earlier identified rosin allergen, 15-hydroperoxyabietic acid (15-HPA). The allergens were detected as their methyl esters. Experimental sensitization and cross reactivity of oxidation products of resin acids were studied. A pattern of cross reactivity was observed which indicates that the hydroperoxide of abietic acid (15-HPA) may react to form a complete antigen via two different routes. One route seems to be via the formation of epoxides which then react with skin protein to form the complete antigen, and the other, via radical formation due to cleavage of the peroxide bond. The radical formed may then react with skin protein, so producing the complete antigen. Few other studies have shown results indicating the formation of several antigens from one hapten. PMID- 9868508 TI - Germ-cell tumors of the mediastinum. AB - Germ-cell tumors occurring primarily in the mediastinum have been well recognized in the literature for many years. Unfortunately, most of what has been written on these tumors has been in the form of case reports and small series of cases, thus precluding a better understanding of their clinical and demographic features, as well as of their biologic behavior in this particular location. In addition, mediastinal germ-cell tumors not only have never had a standardized nomenclature, but they also have lacked the benefit of a clinical staging scheme for assessment of prognosis. Another important aspect of these tumors that has required attention is the frequency with which teratomatous lesions are associated with malignant components in the mediastinum. Comprehensive studies of mediastinal germ-cell tumors to determine whether these lesions recapitulate the distribution, histopathologic features, and biologic behavior of their counterparts in other areas, such as the gonads, have been lacking for many years. In this article, we review our experience with a large series of cases of primary mediastinal germ cell tumors and examine recent concepts and advances in our understanding of these unusual and diagnostically challenging neoplasms. PMID- 9868509 TI - How many types of biliary hamartomas and adenomas are there? PMID- 9868510 TI - Hepatosplenic gamma delta T-cell lymphoma--not just alphabet soup. PMID- 9868511 TI - Post-transplantation lymphoproliferative disorder versus acute liver rejection: do we need a scoring system? PMID- 9868512 TI - In search of a positive immunohistochemical marker for mesothelioma: an update. PMID- 9868513 TI - Calretinin: a novel tool for diagnostic immunohistochemistry. PMID- 9868514 TI - Application of immunocytochemistry in the diagnosis of soft tissue sarcomas: a review and update. AB - The diagnosis and classification of soft tissue sarcomas has been a source of difficulty for surgical pathologists. This has been largely attributable to a significant degree of overlap in histologic features among different types of soft tissue sarcomas. The immunohistochemical identification of various cell and tissue markers specific for certain avenues of cell differentiation has permitted more accurate diagnosis and classification of these tumors. PMID- 9868515 TI - Telomerase activity in cancer: a magic bullet or a mirage? AB - Telomerase is an important enzyme required for maintenance of chromosome ends (telomeres) during cell division. Since elevated activity of telomerase has been selectively demonstrated in a large number of human cancers, it has generated considerable excitement as a potential cancer marker, a prognostic factor, and a target of possible anticancer strategy. This article details diagnostic, prognostic, and therapeutic prospects linked to the measurement of telomerase activity and the growing complexity of its regulation and function. PMID- 9868516 TI - Juvenile polyp--a premalignant lesion? PMID- 9868517 TI - Liver cell dysplasia and other atypical lesions: new insights and applications. PMID- 9868518 TI - Transitional cell metaplasia and neoplasia in the female genital tract: an update. PMID- 9868519 TI - Effects of leuprolide acetate on treatment of leiomyomata--clues to mechanisms of action. PMID- 9868520 TI - Leukoplakia: still a gallimaufry or is progress being made?--A review. AB - Leukoplakia as a medical term has been encumbered by a hodgepodge of synonyms and interpretations. It also has been a term associated with strong emotions with respect to an implied malignant potential. It now is regarded as potentially malignant, with varying degrees of risk for transformation. Leukoplakia at its major anatomic site of occurrence, the oral cavity, recently has been redefined, classified, and staged as a preamble to a more rational approach to the clinicopathologic study of leukoplakia. These changes are presented, as are current assessments of proliferative verrucous leukoplakia, verrucous hyperplasia, palatal white lesions, effects of smokeless tobacco on the oral mucosa, hairy leukoplakia, and lichen planus. PMID- 9868521 TI - The importance of heterogeneity in tumor pathology. AB - Heterogeneity of cell, tissue, and tumor genotype, form, and function confounds clinicopathologic research and therapeutic strategies. Its significance and importance usually are underestimated and underreported, and there is no consistent statistical or biomathematical framework for its interpretation. New technologies for the quantitative study of pathology specimens and a growing awareness of the clinical importance of biodiversity within individual tumors and populations of tumors herald new approaches to the problems posed by dynamic structural and functional complexity in human tumors. PMID- 9868522 TI - Inclusion body myositis--a review. AB - Inclusion body myositis (IBM), a sporadic inflammatory myopathy, is the most frequently occurring progressive myopathy in adults older than 55 years. It more commonly affects men and usually is clinically and pathologically distinguishable from dermatomyositis or polymyositis. Muscle biopsy specimens will display inflammation in virtually all cases of sporadic IBM, along with rimmed vacuoles accompanied by the accumulation of beta-amyloid and other substances similar to those found in the degenerating neurons of Alzheimer's disease. The similarities between IBM and other inflammatory myopathies may contribute to its low level of diagnosis by pathologists. The proper recognition of IBM is important because, unlike other inflammatory myopathies, IBM is unresponsive to anti-inflammatory drugs. PMID- 9868523 TI - A new kind of Alzheimer's disease plaque. PMID- 9868524 TI - Molecular diagnosis of cancer and micrometastases. PMID- 9868526 TI - Transitional cell metaplasia. PMID- 9868525 TI - RET protooncogene analysis in the diagnosis of medullary thyroid carcinoma and multiple endocrine neoplasia type II. PMID- 9868527 TI - Macromolecular cytosolic delivery: cell membranes as the primary obstacle. AB - The "evolution" of a thing, a custom, an organ is thus by no means its progressus toward a goal, even less a logical progressus by the shortest route and with the least expenditure of force, but a succession of more or less profound, mutually independent processes of subduing, plus the resistances they encounter, the attempts at transformation for the purpose of defense and reaction, and the results of successful counteractions. The form is fluid, but the "meaning" is even more so. PMID- 9868528 TI - P62 and the sequestosome, a novel mechanism for protein metabolism. AB - In addition to selecting proteins for degradation by the 26S proteasome, ubiqitination appears to serve other regulatory functions, including for endosomal/lysosomal targeting, protein translocation, and enzyme modification. Currently, little is known how multiubiquitin chains are recognized by these cellular mechanisms. Within the 26S proteasome, one subunit (Mcb1/S5a) has been identified that has affinity for multiubiquitin chains and may function as a ubiquitin receptor. We recently found that a non-proteasomal protein p62 also preferentially binds multiubiquitin chains and forms a novel cytoplasmic structure "sequestosome" which serves as a storage place for ubiquitinated proteins. In the present manuscript, the role and regulation of p62 in relation to the sequestosomal function will be reviewed. PMID- 9868529 TI - Chemokine receptors in HIV-1 and SIV infection. AB - Seven transmembrane segment (7TMS) receptors for chemokines and related molecules have been demonstrated to be essential, in addition to CD4, for HIV and SIV infection. The beta-chemokine receptor CCR5 is the primary, perhaps sole, coreceptor for HIV-1 during the early and chronic phases of infection, and supports infection by most primary HIV-1 and many SIV isolates. Late-stage primary and laboratory-adapted HIV-1, HIV-2, and SIV isolates can use other 7TMS receptors. CXCR4 appears especially important in late-stage HIV infection; several related receptors can also be used. The specificity of SIV viruses is similar. Commonalities among these receptors, combined with analyses of mutated molecules, indicate that discrete, conformationally-dependent sites on the chemokine receptors determine their association with the third variable and conserved regions of viral envelope glycoproteins. These studies are useful for elucidating the mechanism and molecular determinants of HIV-1 entry, and of inhibitors to that entry. PMID- 9868530 TI - Overexpression of Rb and E2F-1 in ataxia-telangiectasia lymphocytes. AB - AT cells exhibit defective cell cycle regulation following DNA damage. Previous studies have shown that induction of p53 and p21 proteins are delayed in response to ionizing radiation, resulting in the failure of G1/S checkpoint in AT cells. In this study, further investigation of the molecular mechanisms underlying G1/S phase progression in AT cells was conducted. Exponentially growing normal and AT cells were exposed to 2 Gy of ionizing radiation and the expression levels and functional activities of Rb and E2F-1 proteins were determined. We observed overexpression of hyperphosphorylated Rb and E2F-1 proteins in AT cells, which was unaffected post-irradiation. Furthermore, gel shift assays showed that E2F-1 DNA binding was constitutive in AT cells, whereas it was inhibited in control cells following exposure to ionizing radiation. The data suggests that abnormalities in the function of Rb and E2F-1 proteins may also be responsible for the failure of AT cells to arrest in the G1/S checkpoint in response to DNA damage. PMID- 9868531 TI - Biphasic release characteristics of dual drug-loaded alginate beads. AB - The dual drug-loaded alginate beads simultaneously containing drug in inner and outer layers were prepared by dropping plain (single-layered) alginate beads into CaCl2 solution. The release characteristics were evaluated in simulated gastric fluid for 2 h followed by intestinal fluids thereafter for 12 h. The surface morphology and cross section of dual drug-loaded alginate beads was also investigated using scanning electron microscope (SEM). The poorly water-soluble ibuprofen was chosen as a model drug. The surface of single-layered and dual drug loaded alginate beads showed very crude and roughness, showing aggregated particles, surface cracks and rough crystals. The thickness of dual drug-loaded alginate beads surrounded by outer layer was ranged from about 57 to 329 microns. The distinct chasm between inner and outer layers was also observed. In case of single-layered alginate beads, the drug was not released in gastric fluid but was largely released in intestinal fluid. However, the release rate decreased as the reinforcing Eudragit polymer contents increased. When the plasticizers were added into polymer, the release rate largely decreased. The release rate of dual drug loaded alginate beads was stable in gastric fluid for 2 h but largely increased when switched in intestinal fluid. The drug linearly released for 4 h followed by another linear release thereafter, showing a distinct biphasic release characteristics. There was a difference in the release profiles between single layered and dual drug-loaded alginate beads due to their structural shape. However, this biphasic release profiles were modified by varying formulation compositions of inner and outer layer of alginate beads. The release rate of dual drug-loaded alginate beads slightly decreased when the outer layer was reinforced with Eudragit RS100 polymers. In case of dual drug-loaded alginate beads with polymer-reinforced outer layer only, the initial amount of drug released was low but the initial release rate (slope) was higher due to more swellable inner cores when compared to polymer-reinforced inner cores. The current dual drug-loaded alginate beads may be used to deliver the drugs in a time dependent manner. PMID- 9868532 TI - Determination of aloesin in plasma by high-performance liquid chromatography as fluorescent 9-anthroyl derivative. AB - A sensitive high-performance liquid chromatographic (HPLC) method for the determination of aloesin in plasma was developed. After solid-phase extraction from plasma and derivatization of aloesin and compound AD-1, which was prepared from aloesin as a internal standard, with 9-anthroylnitrile in the presence of quinuclidine, the derivatives were separated on a inertsil ODS-3 column using acetonitrile/methanol/water (3:1:6) as a mobile phase, and detected fluorimetrically at 460 nm with excitation at 360 nm. The detection limit of aloesin was 3.2 ng/ml in plasma (S/N = 3). PMID- 9868533 TI - NO/cGMP pathway is involved in exocrine secretion from rat pancreatic acinar cells. AB - The enzyme responsible for the synthesis of nitric oxide (NO) from L-arginine in mammalian tissues is known as nitric oxide synthase (NOS) (EC.1.14.13.39). In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat pancreatic acinar cells. Treatment of rat pancreatic acinar cells with cholecystokinin-octapeptide (CCK-OP) resulted in an increase in the arginine conversion to citrulline, the amount of NOx, the release of amylase, and the level of cGMP. Especially, CCK-OP-stimulated increase of arginine to citrulline transformation, the amount of NOx and cGMP level were completely counteracted by the inhibitor of NOS, NG-monomethyl-L-arginine (MMA), by contrast, that of amylase release was partially reduced. Furthermore, MMA-induced decrease of NOS activity and amylase release showed dose-dependent pattern. The data on the time course of CCK-OP-induced citrulline formation and cGMP rise indicate that NOS and guanylate cyclase were activated by treatment of CCK-OP. However, the mechanism of agonist-stimulated guanylate cyclase activation in acinar cells remains unknown. Therefore, activation of NOS is one of the early events in receptor-mediated cascade of reactions in pancreatic acinar cells and NO, not completely, but partially mediate pancreatic enzyme exocrine secretion. PMID- 9868534 TI - Intracellular Ca2+ mediates lipoxygenase-induced proliferation of U-373 MG human astrocytoma cells. AB - The role of intracellular Ca2+ in the regulation of tumor cell proliferation by products of arachidonic acid (AA) metabolism was investigated using U-373 MG human astrocytoma cells. Treatment with nordihydroguaiaretic acid (NDGA), a lipoxygenase (LOX) inhibitor, or caffeic acid (CA), a specific 5-LOX inhibitor, suppressed proliferation of the tumor cells in a dose-dependent manner. However, indomethacin (Indo), a cyclooxygenase (COX) inhibitor, did not significantly alter proliferation of the tumor cells. At anti-proliferative concentrations, NDGA and CA significantly inhibited intracellular Ca2+ release induced by carbachol, a known intracellular Ca2+ agonist in the tumor cells. Exogenous administration of leukotriene B4 (LTB4), an AA metabolite of LOX pathway, enhanced proliferation of the tumor cells in a concentration-dependent fashion. In addition, LTB4 induced intracellular Ca2+ release. Intracellular Ca2+ inhibitors, such as an intracellular Ca2+ chelator (BAPTA) and intracellular Ca(2+)-release inhibitors (dantrolene and TMB-8), significantly blocked the LTB4 induced enhancement of cell proliferation and intracellular Ca2+ release. These results suggest that LOX activity may be critical for cell proliferation of the human astrocytoma cells and that intracellular Ca2+ may play a major role in the mechanism of action of LOX. PMID- 9868535 TI - Ofloxacin resistance mechanism in PA150 and PA300-clinical isolates of Pseudomonas aeruginosa in Korea. AB - Five hundred and seventy clinical strains of Pseudomonas aeruginosa were isolated from August 1993 to August 1994 in Korea and screened for their resistance to ciprofloxacin, norfloxacin, and ofloxacin. Among these, two P. aeruginosa strains (PA150 and PA300) were selected based on their strong resistance (MICs > 50 micrograms/ml) to all three quinolones. The susceptible strain as well as two resistant strains had proton gradient-dependent efflux system. Efflux system in PA300 showed different specificities to ofloxacin and ciprofloxacin while PA 150 had less permeability for ofloxacin. Ofloxacin had a less inhibitory action on DNA synthesis in permeabilized cells of PA150 and PA300 than 1771M. When quinolone resistance determining region (QRDR) in gyrA was sequenced, PA300 had one missense mutation, Asn 116Tyr, which was newly reported in this work. The results showed that PA150 became ofloxacin resistant by reduced ofloxacin accumulation due to the existence of efflux system and low permeability, while resistance of PA300 was due to the efflux system and a mutation in QRDR of gyrA the target site of quinolone. PMID- 9868536 TI - Metabolism of an anionic fluorescent dye, 1-anilino-8-naphthalene sulfonate (ANS) by rat liver microsomes. AB - The present study was designed to examine the metabolism of 1-anilino-8 naphthalene sulfonate (ANS), an anionic compound which is transported into liver via "multispecific organic anion transporter", with rat hepatic microsomes. TLC analysis indicated that the fluorescent metabolites were not produced to a measurable extent, which made it possible to assess the ANS metabolism by measuring the fluorescence disappearance. The metabolism of ANS was remarkably inhibited by the presence of SKF-525A as well as by the substitution of O2 by CO gas. ANS metabolism by microsomes also required NADPH as a cofactor. These results indicated that the microsomal monooxygenase system might be mainly responsible for the ANS metabolism. The maximum velocity (Vmax) and Michaelis constant (K(m)) were calculated to be 4.3 +/- 0.2 nmol/min/mg protein and 42.1 +/ 2.0 microM, respectively. Assuming that 1 g of liver contains 32 mg of microsomal protein, the Vmax value was extrapolated to that per g of liver (Vmax'). The intrinsic metabolic clearance (CLint) under linear conditions calculated from this in vitro metabolic study was 3.3 ml/min/g liver, being comparable with that (3.0 ml/min/g liver) calculated by analyzing the in vivo plasma disappearance curve in a previous study. Furthermore, the effects of other organic anions on the metabolism of ANS were examined. Bromophenolblue (BPB) and rose bengal (RB) competitively inhibited the metabolism of ANS, while BSP inhibited it only slightly. The inhibition constant (Ki) of BPB (6 microM) was much smaller than that of RB (200 microM). In conclusion, the microsomal monooxygenase system plays a major role in the metabolism of ANS, and other unmetabolizable organic anions (BPB and RB) compete for this metabolism. PMID- 9868537 TI - Reduction of radiation-induced chromosome aberration and apoptosis by dithiothreitol. AB - We have examined in vitro and in vivo radioprotective effects of a well-known thiol-containing compound, dithiothreitol (DTT). The treatment of both 0.5 and 1 mM of DTT significantly increased clonogenic survival of gamma-ray irradiated Chinese hamster (V79-4) cells. In order to investigate the possible radioprotective mechanism of DTT, we measured gamma-ray induced chromosome aberration by micronucleus assay. In the presence of 0.5 mM or 1 mM DTT, the frequencies of micronuclei were greatly reduced in all dose range examined (1.5-8 Gy). Slightly higher reduction in micronucleus formation was observed in 1 mM DTT treated cells than in 0.5 mM DTT-treated cells. In addition, incubation with both 0.5 and 1 mM of DTT prior to gamma-ray irradiation reduced nucleosomal DNA fragmentation at about same extent, this result suggests that treatment of DTT at concentrations of 0.5 and 1 mM reduced radiation-induced apoptosis. In vivo experiments, we also observed that DTT treatment reduced the incidence of apoptotic cells in mouse small intestine crypts. In irradiated control group 4.4 +/- 0.5 apoptotic cells per crypt were observed. In DTT-administered and irradiated mice, only 2.1 +/- 0.4 apoptotic cells per crypt was observed. In vitro and in vivo data obtained in this study showed that DTT reduced radiation induced damages and it seems that the possible radioprotective mechanisms of action of DTT are prevention of chromosome aberration. PMID- 9868538 TI - Growth inhibitory activities of kalopanaxsaponins A and I against human pathogenic fungi. AB - Antifungal activities of the compounds isolated from Kalopanax pictus against representative fungi of dermatomycosis were investigated using paper disc diffusion method. It was found that kalopanaxsaponins A and I were effective in inhibiting the growth of Candida albicans KCTC 1940 and Cryptococcus neoformans KCTC 7224 with minimum inhibitory concentration (MIC) of 25 micrograms/ml. It showed that antifungal activity of both compounds have strong selectivity against the fungi of dermatomycosis. PMID- 9868539 TI - Purification and characterization of GTP cyclohydrolase I from Streptomyces tubercidicus, a producer of tubercidin. AB - GTP cyclohydrolase I catalyzing the first reaction in the biosynthesis of pterin moiety of folic acid in bacteria, was purified from Streptomyces tubercidicus by at least 203-fold with a yield of 32% to apparent homogeneity, using ammonium sulfate fractionation, DEAE-cellulose, Sepharose CL-6B, and hydroxylapatite column chromatography. The molecular weight of the native enzyme was estimated to be 230,000 daltons by gel permeation chromatography. The purified enzyme gave a single band on sodium dodesyl sulfate-polyacrylamide gel electrophoresis and its molecular weight was apparently 58,000 daltons. These results indicate that the enzyme consists of four subunits with the same molecular weight. The K(m) and Vmax values for GTP of the purified enzyme were determined to be 80 microM and 90 nmol/min (mg protein), respectively. The optimum pH and temperature for the enzyme reaction were pH 7.5-8.5 and 40-42 degrees C, respectively. Coenzyme or metal ion was not required for the enzyme activity. The enzyme activity was inhibited by most divalent cations, while it was slightly activated by potassium ion. In case of nucleotides, CTP, GMP, GDP, and UTP inhibited enzyme activity, among which GDP exhibited the strongest inhibitory effect. PMID- 9868540 TI - Pharmacokinetics of propentofylline and the quantitation of its metabolite hydroxypropentofylline in human volunteers. AB - Propentofylline (PPF, 3-methyl-1-(5-oxohexyl)-7-propylxanthine) has been reported to be effective for the treatment of both vascular dementia and dementia of the Alzheimer type. The pharmacological effects of PPF may be exerted via the stimulation of nerve growth factor, increased cerebral blood flow, and inhibition of adenosine uptake. The objectives of this experiment are to determine the kinetic behavior of PPF, to identify, and to quantify its metabolite in human. Blood samples were obtained from human volunteers following oral administration of 200 mg of PPF tablets. For the identification and quantification of the metabolite, 3-methyl-1-(5-hydroxyhexyl)-7-propylxanthine (PPFOH), PPFOH was synthesized and identified by gas chromatography/mass spectroscopy (GC/MS) and 1H nuclear magnetic resonance spectroscopy. The molecular weight of synthesized metabolite is 308 dalton. The PPF and PPFOH in plasma were extracted with diethyl ether and identified by electron impact GC/MS. The plasma concentrations of PPF and PPFOH were determined by gas chromatography/nitrogen phosphorus detector in plasma and their pharmacokinetic parameters were determined. The mean half-life of PPF was 0.74 hr. The areas under the curve (AUCs) of PPF and PPFOH were 508 and 460 ng.hr/ml, respectively. Cmax of PPF was about 828.4 ng/ml and the peak concentration was achieved at about 2.2 hr (Tmax). These results indicate that PPF is rapidly disappeared from blood due to extensive metabolism into PPFOH. PMID- 9868541 TI - Anticomplementary activity of stilbenes from medicinal plants. AB - The anticomplementary activity of stilbenes from medicinal plants in Korea was investigated in vitro. 3,5-Dihydroxy-4'-methoxystilbene (3) was most potent with IC50 value of 1.5 x 10(-4) M followed by rhapontigenin (4), oxyresverastrol (2), 2,3,4',5-tetrahydroxystilbene-2-O-beta-glucoside (9), rhaponticin (8), resverastrol (1), and piceid (7). The activity was found to be increased by a methylation on a hydroxy group of C-4' of 1, but decreased by further methylation on hydroxy groups of C-3 and C-5 and glucosylation on any hydroxy group of 1. Addition of hydroxy group on C-2' of 1 or C-3' of 3 was little affected on the anticomplementary activity but the activity was increased by O-glucosylation on C 2 of 1. PMID- 9868542 TI - Possible implication for an indirect interaction between basic fibroblast growth factor and (Na,K)ATPase. AB - The (Na,K)ATPase is responsible for generating the ionic gradients and membrane potentials by the exchange of intracellular Na+ for K+. It has been recently shown that (Na,K)ATPase is involved in the exocytic pathway of basic fibroblast growth factor (bFGF), although it is not known that bFGF is secreted to the outside of cell through direct interaction with (Na,K)ATPase. To understand the role for (Na,K)ATPase in the secretory pathway of bFGF, we have sought to identify the cytoplasmic domains of the alpha 1 isoform of (Na,K)ATPase interacting with bFGF by yeast two-hybrid system. We have also investigated the interaction between the alpha 2 isoform of (Na,K)ATPase and bFGF to find out whether the interaction is isoform-specific. We found that none of the cytoplasmic domains of (Na,K)ATPase isoforms interacted with bFGF. The result suggests that the interaction between bFGF and (Na,K)ATPase might be indirect, thus requiring other proteins which are involved in the formation of protein complexes for the interaction, although we cannot exclude the possibility that the interaction requires the element of the whole alpha subunit structure that was not present in the isolated alpha subunit cytoplasmic domains. PMID- 9868543 TI - Nucleotide and deduced amino acid sequences of rat myosin binding protein H (MyBP H). AB - The complete nucleotide sequence of the cDNA clone encoding rat skeletal muscle myosin-binding protein H (MyBP-H) was determined and amino acid sequence was deduced from the nucleotide sequence (GenBank accession number AF077338). The full-length cDNA of 1782 base pairs(bp) contains a single open reading frame of 1454 bp encoding a rat MyBP-H protein of the predicted molecular mass 52.7 kDa and includes the common consensus 'CA__TG' protein binding motif. The cDNA sequence of rat MyBP-H show 92%, 84% and 41% homology with those of mouse, human and chicken, respectively. The protein contains tandem internal motifs array (-FN III-Ig C2-FN III-Ig C2-) in the C-terminal region which resembles to the immunoglobulin superfamily C2 and fibronectin type III motifs. The amino acid sequence of the C-terminal Ig C2 was highly conserved among MyBPs family and other thick filament binding proteins, suggesting that the C-terminal Ig C2 might play an important role in its function. All proteins belonging to MyBP-H member contains 'RKPS' sequence which is assumed to be cAMP- and cGMP-dependent protein kinase A phosphorylation site. Computer analysis of the primary sequence of rat MyBP-H predicted 11 protein kinase C (PKC) phosphorylation site, 7 casein kinase II (CK2) phosphorylation site and 4 N-myristoylation site. PMID- 9868544 TI - Hepatoprotective activity of scopoletin, a constituent of Solanum lyratum. AB - Scopoletin (7-hydroxy-6-methoxycoumarin), a coumarin, was isolated from the aerial part of Solanum lyratum Thunb. by the activity-guided fractionation employing carbon tetrachloride-intoxicated primary cultured rat hepatocytes as a screening system. Its hepatoprotective activity was first evaluated by measuring the release of glutamic pyruvic transaminase and sorbitol dehydrogenase from carbon tetrachloride-intoxicated rat hepatocytes into the culture medium. Scopoletin significantly reduced the releases of glutamic pyruvic transaminase and sorbitol dehydrogenase from the carbon tetrachloride-intoxicated primary cultured rat hepatocytes by 53% and 58%, respectively, from the toxicity in a dose-dependent manner over concentration ranges of 1 microM to 50 microM. Further studies revealed that at the concentration of 10 microM, scopoletin significantly preserved glutathione content by 50% and the activity of superoxide dismutase by 36% and also inhibited the production of malondialdehyde to the degree as seen in the control. PMID- 9868545 TI - A novel synthesis of heterocyclic compounds containing coumarin moiety of potential antimicrobial activity. AB - The chemical behaviours of 4-methyl-2-oxo-2H-benzopyran-7-yl oxoacetyl hydrazine (2) towards some different reagents such as anhydride compounds, aromatic aldehydes, carbon disulphide, and nitrous acid yielded the corresponding pathalazine derivatives (3, 4, 5), hydrazone derivative (6), 1,3,4-oxadiazole derivative (7, 8, 9) and acid azide (10) respectively. Treatment of 10 with absolute alcohols, amines and ethyl amino acid ester gave the corresponding carbamate derivative (11), substituted urea derivative (12) and ethyl substituted alkyl acetate (13) respectively. The biological activity of some synthesized compounds was evaluated. PMID- 9868546 TI - Topoisomerase I inhibitors from the Streptomyces sp. strain KM86-9B isolated from a marine sponge. AB - The crude extract of Streptomyces sp. strain KM86-9B, isolated from a marine sponge, displayed significant inhibition on topoisomerase I activity. Investigation of the causative components by bioactivity-directed fractionation resulted in the isolation of a series of iso- and anteiso-fatty acids. PMID- 9868547 TI - Screening and isolation of antibiotic resistance inhibitors from herb materials IV-resistance inhibitors from Anetheum graveolens and Acorus gramineus. AB - The hexane fractions from methanolic extracts of Anetheum graveolens L. (Umbelliferae) and Acorus gramineus Soland. (Araceae), revealed potent inhibitory activities against the resistance of multi-drug resistant Staphylococcus aureus SA2 when combined with ampicillin (Am) or chloramphenicol (Cm). As active principles, carvone and the liquid mixture containing carvone from Anetheum graveolens L. and a liquid mixture mainly consisting of benzoic acid phenylmethyl ester (benzyl benzoate) from Acorus gramineus Soland, were identified. They showed resistance inhibition at the level of 20-50 micrograms/ml when combined with 100 or 50 micrograms/ml of Am or Cm, respectively. PMID- 9868548 TI - 6-(1-alkenoyloxyalkyl)-5,8-dimethoxy-1,4-naphthoquinone derivatives:synthesis and evaluation of antitumor activity. AB - Thirty six 5,8-dimethoxy-1,4-naphthoquinone derivatives, which bear unsaturated alkyl side chain with ester bond, were synthesized and tested cytotoxic activity on L1210 cells and antitumor activity against ICR mice bearing S-180 cells. It could be recognized that the cytotoxicities of naphthoquinones with R1 being methyl and propyl (IV1-24) were not enhanced by replacing the alkanoyls with alkenoyls. In contrast, the introduction of the alkenoyl moieties on the compounds with R1 = hexyl (IV25-36) resulted in the enhancement of their cytotoxicities. Replacement of alkanoyl group with an alkenoyl group generally increased the T/C value of the mice bearing S-180 cells. PMID- 9868549 TI - Synthesis of praziquantel derivatives and their in vitro activity against adult Clonorchis sinensis. AB - Several praziquantel derivatives have been prepared by the acylation of compound 5, and examined on their biological activity in vitro a against adult Clonorchis sinensis collected from rabbits infected with metacercariae which was isolated from Pseudorasbora parva, a second intermediate host, captured in Nakdong river in Korea. PMID- 9868550 TI - Synthesis and in vitro cytotoxicity of 3- or 4-dialkylaminomethyl-1 azaanthraquinones. AB - Six 3-dialkylaminomethyl-1-azaanthraquinones and five 4-dialkylaminomethyl-1 azaanthraquinones were synthesized and evaluated in vitro cytotoxicity against four human cancer cell lines. The compounds retained much of their cytotoxic activity against the multi-drug-resistant cell line (KB-V-1) as shown by resistance index. PMID- 9868551 TI - Synthesis and biological evaluation of novel 2-[substituted acetyl]-amino-5-alkyl 1,3,4-thiadiazoles [corrected]. AB - Sixteen novel 2-substituted acetyl amino-5-alkyl-1,3,4-thiadiazole were synthesized and screened for their pharmacological activities. A few of the compounds namely 11, 12 and 16 showed anti-inflammatory activities comparable to phenylbutazone. Compound 12 also showed significant non-specific spasmolytic activity. Diuretic activity of compound 15 at a dose level of 90 mg/kg p.o. was two fold higher compared to 50 mg/kg p.o. of furosemide. Comparable diuresis was also produced by compounds 9, 10 and 16. PMID- 9868552 TI - The effect of N-alkyloxycarbonyl group on the anticonvulsant activities of N alkyloxycarbonyl-alpha-amino-N-methylsuccinimides. AB - In connection with the development of new anticonvulsant agents with a broad spectrum, we found that N-Cbz-alpha-amino-N-alkylsuccinimides showed significant anticonvulsant activities, and the pharmacological activities of these compounds were dependent on their stereochemistry and N-substituted alkyl group. These results prompted us to define the effects of other functional group on the anticonvulsant activities of these compounds. Therefore a series of N alkoxycarbonyl-alpha-amino-N-methylsuccinimide were prepared from N-Cbz-aspartic acid and were evaluated with their anticonvulsant activities against the MES and PTZ tests, in order to define the effect of N-substituted alkoxy carbonyl group with the anticonvulsant activities. From these studies, it was found that all the tested N-alkoxycarbonyl-alpha-amino-N-methylsuccinimides exhibited significant anticonvulsant activities in the PTZ test and were not active in the MES test. The most active compound in the PTZ test was (S) N-ethoxycarbonyl-alpha-amino-N methyl-succinimide. We found that the pharmacological activities in the PTZ test were dependent on their N-alkoxycarbonyl groups. They follow as such; The order of anticonvulsant activities for (R) series as evaluated by ED50 was N phenoxycarbonyl = N-4-nitrobenzyloxycarbonyl > N-ethoxycarbonyl > N allyloxycarbonyl > N-tert. butoxycarbonyl compound; For the (S) series N ethoxycarbonyl > N-phenoxycarbonyl > N-allyloxycarbonyl compound. From the above results, it was conceivable that N-substituted alkoxycarbonyl group had certain effects on the anticonvulsant activities of N-alkoxycarbonyl-alpha-amino-N methylsuccinimides. PMID- 9868553 TI - The effect of N-alkyloxycarbonyl group on the anticonvulsant activities of N alkyloxycarbonyl-alpha-aminoglutarimides. AB - In connection with the development of new anticonvulsant agents with a broad spectrum, we reported that N-Cbz-alpha-aminoglutarimides, combining common structures of other anticonvulsants such as N-CO-C-N and cyclic imides in a single molecule, showed significant anticonvulsant activities in the MES (maximal electroshock seizure) and PTZ (pentylenetetrazole induced seizure) tests. In these studies, a series of (R) and (S) N-alkyloxycarbonyl-alpha-aminoglutarimides 7a-7e and 8a-8e, which were substituted with various alkyloxycarbonyl group instead of Cbz group, were prepared from the corresponding (R) and (S) N-Cbz glutamic acid 3 and 4, and were evaluated with their anticonvulsant activities against the MES and PTZ tests, including neurotoxicity, in order to define the effect of N-alkyloxycarbonyl group on the anticonvulsant activities of N alkyloxycarbonyl-alpha-aminoglutarimides. Among them, (S) N-4 nitrobenzyloxycarbonyl-alpha-amino-N-methylglutarimide 8e was the most active in MES (ED50 = 35.6 mg/kg, PI = 2.7) and PTZ tests (ED50 = 15.6, PI = 6.1). Interestingly, (R) and (S) N-4-nitrobenzyloxycarbonyl-alpha-amino-N methylglutarimide 7e and 8e and (R) N-phenoxycarbonyl-alpha-amino-N methylglutrimide 7d showed significant anticonvulsant activities in both the MES and PTZ tests and other compounds showed anticonvulsant activities in only the PTZ test. In addition, it was found that their anticonvulsant activities were dependent on their stereochemistries and N-substituted alkyloxycarbonyl groups. PMID- 9868554 TI - Effects of Brazilin on induction of immunological tolerance by sheep red blood cells in C57BL/6 female mice. AB - Brazilin was examined for its effects on the induction of immunological tolerance. Brazilin was administered to C57BL/6 female mice for 2 consecutive days before the immunization with high dose SRBC (10(9) cells) which can produce immunological tolerance. Delayed type hypersensitivity, IgM plaque forming cells, ConA induced IL-2 production and mitogen- or antigen-induced proliferation of lymphocytes were measured as evaluation parameters. Administration of brazilin prior to immunization could keep the DTH and IL-2 production almost optimally immunized levels. Brazilin also inhibited the elevation of non-specific suppressor cell activity. ConA induced proliferation of splenocytes in high dose SRBC immunized mice was significantly decreased by pretreatment of brazilin. And this might be one of the reason for augmentation of DTH by brazilin. However, IgM plaque forming cells were not affected by the treatment of brazilin. These results indicate that brazilin prevents the induction of immunological tolerance caused by high dose SRBC by suppressing the elevation of suppressor cell activity and by inhibiting the decrease in IL-2 production in C57BL/6 female mice. PMID- 9868555 TI - Effects of Brazilin on the phospholipase A2 activity and changes of intracellular free calcium concentration in rat platelets. AB - Brazilin (7,11b-dihydrobenz[b]indeno[1,2-d]pyran-3,6a,9,10 (6 H)-tetrol) inhibited thrombin-,collagen- and ADP-induced aggregation of washed rat platelets. Thrombin- and collagen-induced ATP release were also inhibited by brazilin in a concentration-dependent manner. Brazilin inhibited the formation of platelet thromboxane A2 caused by thrombin, whereas it had no effect on the prostaglandin D2 formation. Brazilin inhibited [3H]-arachidonic acid liberation from membrane phospholipids of thrombin-stimulated platelets. Brazilin inhibited the rise of intracellular free calcium caused by thrombin. These results indicate that the inhibition of phospholipase (PLA2) activity and [Ca2+]i elevation might be at least a part of antiplatelet mechanism of brazilin. PMID- 9868556 TI - Eurycoma longifolia increases sexual motivation in sexually naive male rats. AB - The aim of this study is to provide evidence on the aphrodisiac property of Eurycoma longifolia Jack. An electric grid was used as an obstruction in the electrical copulation cage in order to determine how much an aversive stimulus the sexually naive male rat for both the treated with E. longifolia Jack and control groups were willing to overcome to reach the estrous receptive female in the goal cage. The intensity of the grid current was maintained at 0.12 mA and this was the intensity in which the male rats in the control group failed to crossover to reach the goal cage. Results showed that E. longifolia Jack continued to enhance and also maintain a high level of both the total number of successful crossovers, mountings, intromissions and ejaculations during the 9 12th week observation period. In conclusion, these results further enhanced and strengthened the aphrodisiac property of E. longifolia Jack. PMID- 9868557 TI - Effect of ginseng total saponin on bovine adrenal tyrosine hydroxylase. AB - Ginseng total saponin (GTS) can modulate dopaminergic activity at both presynaptic and postsynaptic dopamine receptors (Kim et al., 1998). The present study investigated the effect of GTS on the bovine adrenal tyrosine hydroxylase (TH), which catalyze L/tyrosine to DOP. GTS inhibited the bovine adrenal TH by 42.4, 51.5 and 55.3% at concentrations of 40, 80 and 100 micrograms/ml, respectively. The IC50 value of GTS was 77.5 micrograms/ml. GTS exhibited noncompetitive inhibition with a substrate L-tyrosine. The Ki value was 155 micrograms/ml. PMID- 9868558 TI - Iridoids from Teucrium yemense. AB - The aerial parts of Teucrium yemense yielded two iridoid glycosides. Their structures were elucidated by spectral means as teucardosid and 8-O acetylharpagid. PMID- 9868559 TI - A furan derivative from Cornus officinalis. AB - Dimethyltetrahydrofuran cis-2,5-dicarboxylate a furan derivative has been isolated from the fruits of Cornus officinalis, and it was isolated for the first time in the nature. The structure was elucidated by NMR spectroscopic techniques. PMID- 9868560 TI - Effect of betaine administration on metabolism of hepatic glutathione in rats. PMID- 9868561 TI - Regioselective ring opening of steroidal epoxide with hydrides: formation of C(2) and C(3)-deoxyasiatic acid derivatives. PMID- 9868562 TI - Mental illness and violence: an epidemiological appraisal of the evidence. AB - OBJECTIVE: To conduct a critical review of the literature on the matter of mental illness and violence, to examine whether there is enough evidence to establish a causal link, and to provide epidemiological background to measure the risk to the population. METHOD: Based on a computerized search of the literature on mental illness and violence previously conducted for Health Canada, studies in the area were critically reviewed and divided into 3 main categories: 1) studies of criminal and violent behaviour among psychiatric patients, 2) studies of psychiatric illness among offenders (prevalence studies in institutions, analytical studies, and community follow-up of offenders), and 3) epidemiological community-based studies on the issue of mental illness and violence (police citizen encounters, representative samples, and other epidemiological studies). Causality rules and measures of risk were then applied to the evidence elicited. RESULTS: The review of the literature suggests that only a small minority of hospitalized patients, typically those suffering from acute psychotic symptoms, are involved in violent incidents. Formerly hospitalized patients are at a higher risk of committing violence if they are not properly treated and are experiencing threat/control-override psychotic symptoms. Substance abuse disorders significantly raise the risk for violence. Family members are the most at risk of being victimized. CONCLUSION: An association exists between mental illness and violence, but the many covariations that naturally affect the equation between them introduce uncertainties in establishing causality. PMID- 9868563 TI - Hope, pride, and leadership. PMID- 9868564 TI - Confidentiality in crisis: Part I--The duty to inform. AB - OBJECTIVE: To discuss the ethical and legal imperatives to protect third parties from harm in circumstances often believed to be protected by doctor-patient confidentiality. METHOD: A review of recent legislative changes and legal decisions pertaining to a psychiatrist's duty to warn. CONCLUSION: The current legislative and legal climate presents many risks to the concept of doctor patient confidentiality. Psychiatrists must carefully consider any guarantees of confidentiality made to patients and must discuss the limits of confidentiality when obtaining consent for treatment. PMID- 9868565 TI - Confidentiality in crisis: Part II--Confidentiality of treatment records. AB - OBJECTIVE: To discuss the implications of recent legislative changes and court decisions in Canada that have placed the privacy of psychiatric records information at risk. New areas of exposure include client access to clinical information provided by family members, parental access to children's records, and court access to clinical records in civil, criminal, and family law matters. METHOD: A review of recent legislative changes and court decisions pertaining to access to psychiatric records. CONCLUSION: At present, psychiatric records can no longer be regarded as confidential. Recent changes in the concept of privilege of treatment records necessitates several changes in psychiatric practice regarding informed consent to treatment, content of clinical records, and responses to demands for information. PMID- 9868566 TI - Psychiatrists' documentation of informed consent. AB - OBJECTIVE: The present study was undertaken to determine current attitudes and behaviour toward informed consent for antipsychotic medication and documentation of the informed consent process in patient charts. METHOD: Thirty psychiatrists treating a minimum of 10 patients on antipsychotic medication were selected from teaching and nonteaching hospitals. Clinicians completed questionnaires on their behaviour and attitudes regarding documentation of informed consent and antipsychotic medication. Physicians' charts were reviewed to ascertain documentation. RESULTS: Psychiatrists reported sometimes documenting the informed consent process. The chart review revealed that, on average, each psychiatrist had documentation in 23% of charts. Physicians who either were affiliated with a teaching hospital or spent more time reading medical journals were more likely to document the informed consent process. CONCLUSIONS: Physicians who use antipsychotic medication as a treatment in their practice are not routinely documenting the informed consent process in patient records. Physicians should pay more attention to this aspect of record keeping because it is their only record of the consent process. PMID- 9868567 TI - Phase-IV multicentre clinical study of risperidone in the treatment of outpatients with schizophrenia. The RIS-CAN-3 Study Group. AB - OBJECTIVE: Since most clinical trials of atypical antipsychotics have been conducted in hospitalized patients, a Phase-IV, multicentre, 8-week, open-label, flexible-dose study was performed to assess the efficacy and safety of risperidone in outpatients with schizophrenia. METHOD: Three hundred and thirty patients with a Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R) diagnosis of schizophrenia were enrolled at 61 Canadian sites. Upon trial entry, the patients had their neuroleptic and antiparkinsonian drugs discontinued, and treatment with risperidone was initiated at a dose of 2 mg daily, then increased by 2 mg daily on each of the 2 following days until the initial target dose of 6 mg daily was reached on day 3. No further titration was allowed until day 14, after which the dose could be increased or decreased. RESULTS: During the stabilization phase (days 14-56), the dose was unchanged in 44% of the patients, increased in 24%, decreased in 23%, and titrated both up and down in 9% of the patients. In the efficacy-evaluable population (n = 292), treatment with risperidone produced substantial (-26.4) and significant (P = 0.0001) improvement in the total Positive and Negative Syndrome Scale (PANSS) score. At the end of the study (week 8), 85% of patients were classified as clinically improved according to an a priori definition (that is, 20% or more decrease from baseline in total PANSS score). On their last study visit, 75% of patients reported their experience with risperidone as better than their previous neuroleptic therapy. Risperidone was generally well tolerated. The adverse events reported by more than 5% of the patients were insomnia, nausea, headache, somnolence, dizziness, fatigue, anxiety, vomiting, and ejaculation disorder. Seventy-four percent of the reported treatment-related adverse events were recorded during the first 2 weeks of the trial, possibly because of the discontinuation of prior neuroleptic and antiparkinsonian drugs followed by immediate upward titration of risperidone. However, only 8.5% of adverse events were reported to have occurred during week 3, and only 0.8% of adverse events were reported for week 8. Risperidone treatment produced significant improvements over baseline in the incidence and severity of extrapyramidal symptoms (EPS). A slight but statistically significant increase in body weight was observed. CONCLUSIONS: The results of this open label, Phase-IV trial in a large population of outpatients with schizophrenia found that risperidone was superior to the neuroleptics that patients had previously taken in terms of efficacy and severity of EPS. Our results suggest the use of risperidone at lower doses in outpatients with schizophrenia. PMID- 9868568 TI - The adequacy of reporting randomized, controlled trials in the evaluation of antidepressants. AB - OBJECTIVE: To evaluate the adequacy of the reports of methodological and statistical aspects of randomized, controlled trials that evaluate antidepressant medications and the degree to which their results can be used in subsequent metaanalyses. DATA SOURCES: Randomized, controlled trials published in English that compared 2 antidepressant drugs with a placebo were reviewed. Papers were located using Medline and reference lists. DATA EXTRACTION: Each paper was evaluated using a checklist, and 3 summary criteria scores were derived: minimal, ideal, and overall. RESULTS: Only 9 of the 69 papers met the minimal criteria to be included in a metaanalysis (which would report the final sample size and an estimate of the mean and of the standard deviation); and 0 met all of the ideal criteria for reporting clinical trials. Out of a possible 100 points, the mean score for the articles was 51, and the highest score was 80. Scores were not related to the citation impact factor of the journal in which they appeared. CONCLUSIONS: Researchers must become more aware of the criteria for reporting clinical trials, and editors must insist more strenuously that these criteria be satisfied. PMID- 9868569 TI - The relationship of catatonia symptoms to symptoms of schizophrenia. AB - OBJECTIVE: To evaluate the relationships of symptoms of catatonic schizophrenia to 77 symptoms relevant for diagnosing schizophrenia and to socioanamnestic variables. METHOD: Data from a sample of 112 Canadian patients diagnosed with schizophrenia according to the Diagnostic and Statistical Manual of Mental Disorders (DSM-III) were evaluated via phi correlation coefficients. RESULTS: Forty-five (40.2%) of our 112 patients had catatonic symptoms, either at the time of this study or in the past. However, only weak correlations (phi < 0.31) to other symptoms relevant for diagnosing schizophrenia were found, and no significant correlations to socioanamnestic variables were found. CONCLUSION: Symptoms of catatonia appear to be independent of the key symptoms of schizophrenia. PMID- 9868570 TI - [Abuse of psychoactive drugs and social adjustment of psychotic patients]. AB - OBJECTIVE: Is the abuse of psychoactive drugs in psychotic patients linked to social adjustment? METHOD: Fifty-five psychotic men from a detention centre or a psychiatric hospital were assessed with the Social Adjustment Scale (SAS-II) and a French version of the Phillips Rating Scale of Premorbid Adjustment in Schizophrenia. RESULTS: In psychotic patients, the abuse of psychoactive drugs is linked to some indicators of social adjustment and premorbid sexual adaptation. CONCLUSION: Differences were found in some aspects of social functioning, but it is difficult to establish an overall assessment of social adjustment. PMID- 9868571 TI - Adoptee overrepresentation among clinic-referred boys with gender identity disorder. AB - OBJECTIVES: To test the hypothesis that adoptees are overrepresented among a sample of clinic-referred boys with gender identity problems (N = 238). To compare the adoptees and nonadoptees on demographic, behaviour problem, and gender-typed measures. METHOD: The percentage of clinic-referred boys with gender identity problems adopted in the first 2 years of life ("early adoptees") was compared to the base rate of boys adopted in Ontario. Parent-report and behavioural measures were used to compare the early adoptees with "late adoptees" (adopted after the second year of life) and nonadoptees. RESULTS: The percentage of boys with gender identity problems who were early adoptees (7.6%) was significantly higher than the base rate of males adopted in Ontario in the first 2 years of life (1.5%). Both the early and late adoptees were significantly less intelligent than the nonadoptees. The early adoptees also had significantly higher externalizing T scores on the Child Behavior Checklist than did the late adoptees and the nonadoptees. The 3 groups did not differ in the percentage who met the complete Diagnostic and Statistical Manual of Mental Disorders (DSM) criteria for gender identity disorder and on 4 other measures of gender-typed behaviour. CONCLUSION: Adoptees are overrepresented among clinic-referred boys with gender identity problems. The reasons for this finding are not clear but may be accounted for by general risk factors that increase the likelihood of clinical referral or by psychosocial and biological factors associated with adoption. PMID- 9868572 TI - Risperidone for insomnia in PDDs. PMID- 9868573 TI - Fetal alcohol syndrome and seizure disorder. PMID- 9868574 TI - Urinary retention with sertraline, haloperidol, and clonazepam combination. PMID- 9868575 TI - Body image alterations in binge eating disorder. PMID- 9868576 TI - Acute dystonic reaction with trazodone. PMID- 9868577 TI - Aggression, agitation, and mania with olanzapine. PMID- 9868578 TI - Empowering the staff in long-stay wards. PMID- 9868579 TI - Quantifying the cleanliness of glass capillaries. AB - I used capillary rise methods to investigate the lumenal surface properties of quartz (fused silica, Amersil T-08), borosilicate (Corning 7800), and high-lead glass (Corning 0010) capillaries commonly used to make patch pipets. I calculated the capillary rise and contact angle for water and methanol from weight measurements. The capillary rise was compared with the theoretical maximum value calculated by assuming each fluid perfectly wetted the lumenal surface of the glass (i.e., zero contact angle, which reflects the absence of surface contamination). For borosilicate, high-lead, and quartz capillaries, the rise for water was substantially less than the theoretical maximum rise. Exposure of the borosilicate, lead, and quartz capillaries to several cleaning methods resulted in substantially better--but not perfect--agreement between the theoretical maximum rise and calculated capillary rise. By contrast, the capillary rise for methanol was almost identical in untreated and cleaned capillaries, but less than its theoretical maximum rise. The residual discrepancy between the observed and theoretical rise for water could not be improved on by trying a variety of cleaning procedures, but some cleaning methods were superior to others. The water solubility of the surface contaminants, deduced from the effectiveness of repeated rinsing, was different for each of the three types of capillaries examined: Corning 7800 > quartz > Corning 0010. A surface film was also detected in quatz tubing with an internal filament. I conclude that these borosilicate, quartz, and high-lead glass capillaries have a film on the lumenal surface, which can be removed using appropriate cleaning methods. The surface contaminants may be unique to each type of capillary and may also be hydrophobic. Two simple methods are presented to quantitate the cleanliness of glass capillary tubing commonly used to make pipets for studies of biological membranes. It is not known if the surface film is of importance in electrophysiological studies of biological membranes. PMID- 9868580 TI - Cell contraction caused by microtubule disruption is accompanied by shape changes and an increased elasticity measured by scanning acoustic microscopy. AB - The state of crosslinking of microfilaments and the state of myosin-driven contraction are the main determinants of the mechanical properties of the cell cortex underneath the membrane, which is significant for the mechanism of shaping cells. Therefore, any change in the contractile state of the actomyosin network would alter the mechanical properties and finally result in shape changes. The relationship of microtubules to the mechanical properties of cells is still obscure. The main problem arises because disruption of microtubules enhances acto myosin-driven contraction. This reaction and its impact on cell shape and elasticity have been investigated in single XTH-2 cells. Microtubule disruption was induced by colcemid, a polymerization inhibitor. The reaction was biphasic: a change in cell shape from a fried egg shape to a convex surface topography was accompanied by an increase in elastic stiffness of the cytoplasm, measured as longitudinal sound velocity revealed by scanning acoustic microscope. Elasticity increases in the cell periphery and reaches its peak after 30 min. Subsequently while the cytoplasm retracts from the periphery, longitudinal sound velocity (elasticity) decreases. Simultaneously, a two- to threefold increase of F-actin and alignment of stress fibers from the cell center to cell-cell junctions in dense cultures are induced, supposedly a consequence of the increased tension. PMID- 9868581 TI - Subcellular tension fields and mechanical resistance of the lamella front related to the direction of locomotion. AB - Keratocytes derived from the epidermis of aquatic vertebrates are now widely used for investigation of the mechanism of cell locomotion. One of the main topics under discussion is the question of driving force development and concomitantly subcellular force distribution. Do cells move by actin polymerization-driven extension of the lamella, or is the lamella edge extended at regions of weakness by a flow of cytoplasm generated by hydrostatic pressure? Thus, elasticity changes were followed and the stiffness of the leading front of the lamella was manipulated by local application of phalloidin and cytochalasin D (CD). In scanning acoustic microscopy (SAM), elasticity is revealed from the propagation velocity of longitudinal sound waves (1 GHz). The lateral resolution of SAM is in the micrometer range. Using this method, subcellular tension fields with different stiffnesses (elasticity) can be determined. A typical pattern of subcellular stiffness distribution is related to the direction of migration. Cells forced to change their direction of movement by exposure to DC electric fields of varying polarity alter their pattern of subcellular stiffness in relationship to the new direction. The cells spread into the direction of low stiffness and retract at zones of high stiffness. The pattern of subcellular stiffness distribution reveals force distribution in migrating cells; i.e., if a cell moves exactly in a direction perpendicular to its long axis, then the contractile forces are largest along the long axis and decrease toward the short axis. Locomotion in any angle oblique to this axis requires an asymmetric stiffness distribution. Inhibition of actomyosin contractions by La3+ (2 mM), which inhibits Ca2+ influx, reduces cytoplasmic stiffness accompanied by an immediate cessation of locomotion and a change of cell shape. Local release of CD in front of a progressing lamella activates a cell to follow the CD gradient: The lamella thickens locally and is extended toward the tip of the microcapillary. Release of phalloidin stops extension of the lamella, and the cell turns away from the releasing microcapillary. The response to CD is assumed to be the result of local weakening of the cytoplasm due to severing of the actin fibrils. Phalloidin is supposed to stabilize the leading front by inhibition of F-actin depolymerization. These observations are in favor of the assumption that migration is due to an extension of the cell into the direction of minimum stiffness, and they are consistent with the hypothesis that local release of hydrostatic pressure provides the driving force for the flux of cytoplasm. PMID- 9868582 TI - Ruptured fission yeast walls. Structural discontinuities related to the cell cycle. AB - Distributions of rupture sites of fission yeast cells ruptured by glass beads have been related to a new morphometric analysis. As shown previously (Johnson et al., Cell Biophysics, 1995), ruptures were not randomly distributed nor was their distribution dictated by geometry, rather, ruptures at the extensile end were related to cell length just as the rate of extension is related to cell length. The extension patterns of early log, mid-log, late log, and stationary phase cells from suspension cultures were found to approximate the linear growth patterns of Kubitschek and Clay (1986). The median length of cells was found to decline through the log phase in an unbalanced manner. PMID- 9868583 TI - Purinergic receptor-induced changes in paracellular resistance across cultures of human cervical cells are mediated by two distinct cytosolic calcium-related mechanisms. AB - In human cervical (CaSki) cells, extracellular adenosine triphosphate (ATP) induces an acute decrease in the resistance of the lateral intercellular space (RLIS), phase I response, followed by an increase in tight junctional resistance (RTJ), phase II response. ATP also stimulates release of calcium from intracellular stores, followed by augmented calcium influx, and both effects have similar sensitivities to ATP (EC50 of 6 microM). The objective of the study was to determine the degree to which the changes in [Ca2+]i mediate the responses to ATP. 1,2-bis (2-aminophenoxy) ethane-N,N,N1,N1-tetraacetic acid (BAPTA) abrogated calcium mobilization and phase I response; in contrast, nifedipine and verapamil inhibited calcium influx and attenuated phase II response. Barium, La3+, and Mn2+ attenuated phase I response and attenuated and shortened the ionomycin-induced phase I-like decrease in RLIS, suggesting that store depletion-activated calcium entry was inhibited. Barium and La3+ also inhibited the ATP-induced phase II response, but Mn2+ had no effect on phase II response, and in the presence of low extracellular calcium it partly restored the increase in RTJ. KCl-induced membrane depolarization stimulated an acute decrease in RLIS and a late increase in RTJ similar to ATP, but only the latter was inhibited by nifedipine. KCl also induced a nifedipine-sensitive calcium influx, suggesting that acute increases in [Ca2+]i, regardless of mobilization or influx, mediate phase I response. Phase II like increases in RTJ could be induced by treatment with diC8, and were not affected by nifedipine. Biphasic, ATP-like changes in RTE could be induced by treating the cells with ionomycin plus diC8. We conclude that calcium mobilization mediates the early decrease in RLIS, and calcium influx via calcium channels activates protein kinase C and mediates the late increase in RTJ. PMID- 9868585 TI - Therapy and clinical trials. PMID- 9868584 TI - A novel fluorescence chamber for the determination of volume changes in human CaSki cell cultures attached on filters. AB - The objective of the study was to test the hypothesis that, in the cultured human cervical epithelium, CaSki, the effect of calcium mobilizing agents on transepithelial electrical conductance (GTE), is the result of cell volume decrease. CaSki cells attached on filters were loaded with fura-2, and measurements of fluorescence at the isosbestic wavelength 360 nm (excitation/emission [F360/510]) were made in a newly designed fluorescence chamber; this design allowed us also to determine changes in cytosolic calcium ([Ca2+]i). The experimental conditions were similar to those used to measure changes in paracellular permeability in the Ussing chamber, and they enabled us to compare the time-course of changes in [Ca2+]i, in F360/510, and in GTE. Hypertonicity increased, and hypotonicity decreased F360/510 and GTE, without having an effect on [Ca2+]i, and the changes in F360/510 and in GTE correlated linearly. Metabolism, bleaching, and extrusion of intracellular fura-2 were minimal, indicating that the changes in F360/510 reflect changes in dye concentration. Hypertonicity decreased, and hypotonicity increased the size of dispersed CaSki cells, suggesting that osmolarity-induced changes in F360/510 reflect changes in size of the attached cells. Ionomycin increased [Ca2+]i, F360/510, and GTE, but the increases in [Ca2+]i preceded those in F360/510 and GTE. The calcium chelator BAPTA blocked the ionomycin-induced increase in [Ca2+]i, F360/510, and in GTE. Preincubation with 4-acetamido 4'isothiocyanatostilbene-2,2'disulfonic acid (SITS) augmented the ionomycin induced increase in [Ca2+]i, but blocked the increases in F360/510 and in GTE. Pretreatment of cells with hypertonic solution abrogated the increases in F360/510 and in GTE in response to ionomycin, but had little effect on the ionomycin-induced increase in [Ca2+]i. On the basis of these results we suggest that the ionomycin-induced increase in GTE is mediated by [Ca2+]i-dependent chloride secretion and osmotic water loss. PMID- 9868586 TI - Safety and effectiveness of long-term LDL-apheresis in patients at high risk. AB - Since its introduction more than 10 years ago, LDL-apheresis has gained much interest and has proven its clinical utility in patients who cannot be adequately treated by diet and drug therapy alone. A number of clinical studies have clearly demonstrated that regular LDL-apheresis not only favourably influences the progression of coronary artery disease, but also decreases the incidence of cardiovascular events and prolongs survival time of coronary patients at high risk. Both hypercholesterolemia and hyperfibrinogenemia show a high prevalence in heart transplant recipients and seem to cause direct effects on survival time. Heparin-mediated extracorporeal LDL-precipitation-LDL-apheresis has proven to be very successful in this group of patients, which may be caused by the simultaneous removal of LDL, lipoprotein (a) and fibrinogen, and also because LDL apheresis decreases the susceptibility of LDL to oxidation. In addition, there is clear clinical and experimental evidence that LDL-apheresis rapidly improves the endothelial-mediated vasomotion. Not unexpectedly, there are differences in specificity and side-effects between the systems used and these deserve more attention for future routine clinical use. PMID- 9868587 TI - Inhibition of cholesterol absorption by plant sterols for mass intervention. AB - Plant sterols and stanols lower serum cholesterol by inhibiting intestinal absorption of cholesterol. Because of their safety and efficacy, their application for mass intervention is promising. The use of fatty acid esters of stanols is particularly helpful because stanols readily mix with dietary fats in this form and their hypocholesterolemic efficacy is greater than in the free form. PMID- 9868589 TI - Antioxidants and coronary artery disease. AB - Data on the protective role of antioxidants in models of atherosclerosis are only partially confirmed in man. Observational and epidemiological data, as well as randomized trials, provide no clear cut indications, because of positive and disappointing results on the use of antioxidants in cardiovascular protection. Despite the lack of a general consensus, recent data reinforce the concept that the regular intake of antioxidants present in food limits the progression of atherosclerosis. When it is possible to monitor the efficacy of any antioxidant therapy with validated markers of oxidation, the potential influence of vitamins and antioxidants on coronary artery disease may eventually be resolved. PMID- 9868588 TI - Recent data are not in conflict with homocysteine as a cardiovascular risk factor. AB - An elevated level of plasma total homocysteine is an independent, graded and strong risk factor for cardiovascular disease, which shows a strong interactive effect with conventional risk factors. It is a predictor of cardiovascular disease events in high-risk populations, but is weakly associated with risk in healthy individuals. A common polymorphism in the MTHFR gene is a major determinant of the total homocysteine level, but is unrelated to cardiovascular disease risk in most studies. This observation is in accordance with the view that high total homocysteine itself may not be particularly deleterious, but provokes vascular occlusion under conditions predisposing to cardiovascular disease. PMID- 9868590 TI - Follow-up on primary prevention trials. AB - Recent primary prevention trials demonstrated that cardiovascular morbidity and mortality benefits are not accompanied by adverse effects on overall mortality and morbidity in cohorts representing plasma cholesterol concentrations observed in the bulk of coronary artery disease. During the past year, further analyses of the West of Scotland Coronary Prevention Study have indicated that benefit requires a 25% reduction of LDL cholesterol and that such treatment is not very expensive when focussed on selected high-risk individuals. The Air Force/Texas Coronary Artery Prevention Study indicated that benefit is seen in individuals with even lower plasma lipid concentration. Although current treatment with lifestyle and lipid modifying drug management is successful in primary prevention, the unpredictable nature of coronary artery disease and the cost of drugs mitigate against direct application of drug management in persons with relatively low risk, but selective treatment should be undertaken in very high risk settings. Future studies need to examine more specific at risk cohorts, test better targeted lipoprotein modification, test more risk factors and also examine whether changes in vascular function or markers of inflammation will predict a better outcome. PMID- 9868591 TI - Primary and secondary prevention of coronary artery disease: a follow-up on clinical controlled trials. AB - Most probably the decennia of the 1990s will be called the 'statin decennia' in the history of coronary heart disease prevention. Statins are effective, both in primary and secondary prevention of coronary heart disease, in middle-aged and older (< 76 years) men and women, in both diabetics and non-diabetics with coronary heart disease. Statins used in secondary prevention of coronary heart disease significantly reduce the risk of stroke. They also reduce daily attacks of myocardial ischemia. Pathogenetic pathways leading to 'biological plausibilities' of the statins favourable effects are multiple, which explains their rapid (less than 1 year) influence on coronary events. Until the results from new event trials become available, fibrates have very few indications as first line drug therapy in dyslipidemia. They should be considered in combined therapy with statins. The scientific evidence with statins is overwhelming and the question is no longer 'who should we treat?' but 'who can society afford to treat?'. Health economics are indeed pivotal in the use of statins and public health authorities have to find answers according to their resources or innovative strategies, including new aspects in dietary advice (the 'Mediterranean diet'?). PMID- 9868592 TI - Garlic preparations for prevention of atherosclerosis. AB - On the basis of a number of new rigorously designed controlled studies, there is increasingly less evidence for lipid lowering properties of garlic preparations. Many other aspects of garlic drugs, such as direct effects on vessel walls (aortic elasticity, effects of antioxidant properties on early steps in atherosclerosis formation) or anti-platelet aggregation effects, are still awaiting further elucidation in clinical studies. PMID- 9868593 TI - Relevance of the reduction of triglycerides in the prevention of coronary heart disease. AB - Triglyceride metabolism is complex and still poorly understood. Hypertriglyceridemia seems to enhance the risk for the development of coronary heart disease, but it cannot be regarded as a counterpart of hypercholesterolemia as a risk factor. The relevance of reduction of triglycerides in the prevention of coronary heart disease is still unclear. This is related to a number of problems, such as the lack of adequate clinical trials, heterogeneity in the abnormalities of triglyceride metabolism and unknown atherogenic threshold levels for serum triglycerides. PMID- 9868594 TI - Effects of oestrogens and selective oestrogen receptor modulators on serum lipoproteins and vascular function. AB - Epidemiological observations, clinical studies, and basic laboratory research suggest that oestrogen replacement therapy is associated with beneficial cardiovascular effects in postmenopausal women. Oestrogen has a multitude of biological effects that may account for this apparent benefit (which remain to be proven in randomized clinical trials), including favourable effects on the lipid profile, a direct effect on the vascular endothelium with increased nitric oxide bioactivity, and improved fibrinolysis. However, long-term oestrogen therapy increases the risk of breast and endometrial cancers. Raloxifene, a benzothiophene derivative that binds to the oestrogen receptor, is a selective oestrogen receptor modulator, producing oestrogen-agonistic effects in some tissues (liver, bone), and oestrogen-antagonistic effects in others (breast, uterus), and may prove to be an option for women with atherosclerosis and its associated risk factors who might benefit from oestrogen therapy. This review updates the current knowledge of the biological effects of oestrogen and selective oestrogen receptor modulators of potential cardiovascular importance in postmenopausal women. PMID- 9868595 TI - Therapy and clinical trials. PMID- 9868596 TI - Salivary nitrate, nitrite and nitrate reductase activity in relation to risk of oral cancer in Egypt. AB - It has been suggested that nitrate and nitrite may play a role in the etiology of human oral cancer. We investigated whether salivary nitrate and nitrite and the activity of nitrate reductase (NRase) may affect the risk of oral cancer in Egypt, an area with high levels of environmental nitrosating agents. Levels of salivary nitrite (8.3 +/- 1.0 micrograms/ml) and nitrate (44 +/- 3.7 micrograms/ml) and activity of NRase (74 +/- 10 nmol/ml/min) were significantly (P < 0.05) higher in oral cancer patients (n = 42) compared to control Egyptian healthy individuals (n = 40, nitrite = 5.3 +/- 0.3 micrograms/ml, nitrate = 27 +/ 1.2 micrograms/ml, and NRase activity = 46 +/- 4 nmol/ml/min). The adjusted odds ratio (OR) and the 95% confidence intervals (C.I.) for risk of oral cancer, categorized by the levels of salivary nitrate and nitrite and NRase activity, showed a higher cancer risk associated with nitrite > 7.5 micrograms/ml (OR: 3.0, C.I.: 1.0-9.3), nitrite > 40 micrograms/ml (OR: 4.3, C.I.: 1.4-13.3) and NRase activity > 50 nmol/ml/min (OR: 2.9, C.I.: 1.1-7.4). Our findings suggest that increased consumption of dietary nitrate and nitrite is associated with elevated levels of salivary nitrite. Together with the increased activity of salivary NRase, these observations may explain, at least in part, the role of nitrate and nitrite in the development of oral cancer in individuals from an area with a high burden of N-nitroso precursors. PMID- 9868597 TI - Monoclonal antibody recognizing a core epitope on mucin. AB - Monoclonal antibody TH1 (IgM) was prepared by immunizing mice with deglycosylated (TFMSA-treated) cystic fibrosis mucin. TH1 reacted strongly with TFMSA treated cystic fibrosis mucin but not with the fully glycosylated mucin, indicating reactivity with a core mucin epitope. TH1 showed no reactivity with ovine mucin (98% of glycans as sialyl-Tn) but reacted strongly with desialylated ovine mucin, indicating the epitope for this mab was the Tn-antigen (O-linked GalNAc). However, TH1 showed no reactivity with Tn-positive red blood cells, and the binding of TH1 was not inhibited by GalNAc at 2.5 mg/ml, illustrating the importance of the peptide sequence to which the GalNAc is attached. TH1 stained the majority of cancers of the colon, lung, stomach, ovary, breast, and cervix, and the cellular distribution of this antigen in normal tissue suggested reactivity with immature mucin. This antibody appears to be a useful reagent for the detection of immature mucin. PMID- 9868598 TI - Epstein-Barr virus, human papillomavirus, and flow cytometric cell cycle kinetics in nasopharyngeal carcinoma and inverted papilloma among Egyptian patients. AB - It is widely accepted that the Epstein-Barr virus is etiologically associated with the development of nasopharyngeal carcinoma. The human papillomavirus is also associated with inverted papilloma. We used the polymerase chain reaction technique to detect both viruses in both types of tumors. Flow cytometry was also used to study the DNA pattern and proliferative behavior of the tumors in relation to the viruses. EBV was detected in 13/20 (65%) of NPC specimens, and in none of IP (n = 10) or control specimens (n = 10). This indicates the contribution of EBV as an etiologic factor in NPC. Five cases of NPC (25%) were positive for HPV 16, two of them were EBV positive. Four HPV 16 positive cases were found among cases with inverted papilloma, but none among the control cases. Flow cytometry revealed that all NPC, IP, and control samples were diploid except one aneuploid NPC sample. Proliferative capacity (PC) of primary tumors was predictive of tumor recurrence in NPC. Using 13.6% as a cut-off point for PC, we were able to discriminate between high risk and low risk groups with 100% sensitivity and 86% specificity. PC can be used as a baseline prognostic parameter in NPC, making it possible to modify courses of treatment in an attempt to inhibit tumor recurrence. PMID- 9868599 TI - Phosphotyrosine-protein-phosphatase and diabetic disorders. Further studies on the relationship between low molecular weight acid phosphatase genotype and degree of glycemic control. AB - We have studied a new sample of 276 NIDDM patients from the population of Penne (Italy). Comparison of the new data with those of 214 diabetic pregnant women from the population of Rome reported in a previous paper has shown that the pattern of association between low molecular weight acid phosphatase genotype and degree of glycemic control is similar in the two classes of diabetic patients. Among nonobese subjects the proportion of ACP1*A (the allele showing the lowest enzymatic activity) is lower in diabetic patients with high glycemic levels (mean value greater than 8.9 mmol/l) than in diabetic patients with a low glycemic level (mean value less than 8.9 mmol/l). Among obese subjects no significant association is observed between glycemic levels and ACP1. Among nonobese subjects the concentration of f isoform of ACP1 is higher in patients showing a high glycemic level than in patients showing a low glycemic level. No significant difference is observed for s isoform. PMID- 9868600 TI - Disease and the deceased. PMID- 9868601 TI - Physicians' part in moral inquiry. PMID- 9868602 TI - The Capitol shooting. No reflex legislation, no long-term solution. PMID- 9868603 TI - The authority of the clinical ethicist. AB - Mediator? Moral Expert? Or both? "Discourse Ethics" suggests that consensus provides the foundation for defensible moral norms. Thus in building consensus on a moral problem, an ethicist is not just negotiating a compromise but is contributing to the construction of moral rules and principles that have a genuine claim on us. In this way, not only does expertise on a variety of moral positions facilitate mediation, but mediation opens the way to a kind of moral expertise. PMID- 9868604 TI - The nature of ethical expertise. AB - Critics of clinical ethicists sometimes claim that if there were expertise in ethics, then there would have to be objective moral knowledge. They also assume that there would be only one kind of ethics expertise, and that it would be a kind of professional specialization. All three assumptions are mistaken. PMID- 9868605 TI - Resuscitation in hospice. PMID- 9868606 TI - Time for new rules on human subjects research? PMID- 9868607 TI - A world of research subjects. PMID- 9868608 TI - AZT trials and tribulations. PMID- 9868609 TI - Science in the service of healing. PMID- 9868610 TI - Research in developing countries: taking "benefit" seriously. PMID- 9868611 TI - Placebos and HIV. Lessons learned. PMID- 9868613 TI - Improving health care, Part 5: Applying the Dartmouth clinical improvement model to community health. AB - BACKGROUND: Traditional approaches to community health initiatives provide guidance on community mobilization, health assessment, planning, and intervention. Yet direction in how to frame the action steps to implement and measure results is often missing. Many community health initiatives find implementation overwhelming and ineffectual. FRAMEWORK FOR COMMUNITY HEALTH-THE CLINICAL IMPROVEMENT MODEL: The process--outcome methodology of continuous quality improvement (CQI) can translate large community aims into manageable projects. The sequential application of the clinical improvement model and the Community Health Value Compass for measuring outcomes-in state of health, quality of life, satisfaction, and costs-provides a link between data and action, thereby producing accountability for the community health initiative. USING THE CLINICAL IMPROVEMENT MODEL IN TWIN FALLS: Healthy Magic Valley (Twin Falls, Idaho) is the vision for long-term improvement in health status and reduction of health risks for the Southcentral Idaho Health Network. Since 1996 the Twin Falls Community Health Collaborative and SAFE KIDS Coalition have used the Value Compass model and CQI methods to decrease the rate of motor vehicle collisions, serious injuries, and deaths involving teens, while reducing the health, educational, legal, and financial consequences associated with teen-involved motor vehicle collisions. In 1993 the Twin Falls collaborative convened to apply CQI methods to the health of the community. The team has since met periodically to address the issues of community health, using the Dartmouth value compass model since 1996. Each sequential application of the process-outcome CQI framework exposes a blueprint for action and the unfolding of a health improvement strategy. The interventions should affect one or more dimensions of the value compass for teenage driving and motor vehicle collisions. CASE STUDY OF THE CLINICAL IMPROVEMENT MODEL: The motor vehicle death in October 1997 of a high school football player, who was not wearing a seat belt, led to a call to action for injury prevention. Implementation of a local community health initiative on seat belt use started in 1998. A strategy was developed to address implementation of the project among high school teens (for immediate impact) and elementary school children (for long-term impact) and to promote collaboration between the school and the rest of the community. RESULTS: Observed use of seat belts increased from January to September 1998. Data on fatality rates; injury rates; percentages of teens in crashes, of teens injured, and of teen collisions involving use of alcohol; and comprehensive costs are also monitored. DISCUSSION: Once coalitions are built and priorities set, the Dartmouth clinical improvement model presents a method that emphasizes measuring the benefits to the individual members of the community. A portfolio composed of a value compass for each health improvement initiative provides ongoing feedback for guiding subsequent strategic planning by the governing community health network. PMID- 9868614 TI - Failure to immunize the elderly: a systems problem or a statement of personal values? AB - BACKGROUND: Despite the proven efficacy of the influenza vaccine in reducing the risk for pneumonia, hospitalization, and death and the potential savings in costs, most elderly persons do not receive annual immunizations. The study tested the influence of health care delivery system characteristics and individual personal values on the influenza immunization status. METHODS: The study involved a secondary data analysis based on the results of a mailed survey of 3,362 seniors 65 years of age and older enrolled in HealthPartners, a mixed-model health maintenance organization in Minnesota. The three care delivery systems in which respondents were enrolled varied in the intensity and consistency with which they addressed immunization. RESULTS: The immunization rate for this population (77.1%) was higher than the state rate (64%). After controlling for many variables historically known to influence the likelihood of immunization, both care delivery system characteristics and personal values remained significantly associated with immunization status. Elderly individuals getting care in delivery systems with well-developed immunization programs were more likely to be immunized. Those who avoided going to the physician and who practiced risky behaviors such as smoking were less likely to be immunized, regardless of the care delivery system they were enrolled in. DISCUSSION: Managed care can provide a number of system improvements to assist in meeting national health care objectives such as influenza immunization for the elderly. It has reduced some common barriers to immunization, such as cost and access. Yet to achieve the full benefits of a successful influenza immunization program, the role of individual values, as well as implementing systems solutions, needs to be addressed. PMID- 9868615 TI - Accountability for quality in managed care. AB - BACKGROUND: It is often difficult to understand where responsibility lies for monitoring and improving quality in managed care. From 1996 through 1998 a group of individuals convened by the Institute of Medicine's (Washington, DC) National Roundtable on Health Care Quality developed a model of accountability for the quality of care provided by managed care organizations (MCOs). Each of three overarching forms of accountability (professional, market, and regulatory) has a set of tools for imposing accountability and-because accountability relationships are not self-enforcing-sanctions for failures of accountability. PROFESSIONAL ACCOUNTABILITY: Fiduciary relationships in medicine are an essential part of any quality accountability mechanism, and it will be important to maintain the strength of the professional model in the changing health care system. Yet it is not easy to preserve the strength of the professional model in an MCO environment in which professionals are not dominant, and there is likely to be increasing pressure to weaken their autonomy. MARKET ACCOUNTABILITY: The primary assumption of market accountability is that consumers will select options based on perceived value to them and will make new choices based on their information and experience. Market accountability requires choice among competing providers and information to inform choice. In health care, however, individuals rarely have the information they need and often do not have choice. Accountability for quality generally has not been a major feature in contracts. REGULATORY ACCOUNTABILITY: There is a widespread perception of defects in a market-based health care system. Many believe there is a need for a regulatory structure to correct market failures. The use of regulation to impose accountability for quality requires that a regulatory framework, penalties for violations, and effective enforcement mechanisms are all established. PUBLIC GOODS: The model of accountability for quality in managed care does not promote public goods such as education, research, public health, or care for the uninsured. Indeed, the locus of responsibility to the community when markets fail to supply these public goods is controversial. Nevertheless, such responsibility should be considered by MCOs and policy makers. COLLABORATION TO IMPROVE QUALITY OF CARE: Given market-driven models of health care financing and delivery, it might be feasible and desirable to encourage collaboration among MCOs to improve quality, whether at the national or local market level. The health professions in general, and the medical profession in particular, are and must be accountable to society for providing leadership in the development of knowledge about effective medical care, in defining high-quality care, and in advocating for and improving the quality of care. CONCLUSION: Establishing effective accountability for quality involves multiple entities and many different kinds of accountability relationships. The three forms of accountability interact, and all operate at once. PMID- 9868616 TI - Efficacy of silver-coated medical devices. AB - Silver coating of medical devices is believed to prevent device-associated infection. Several in-vitro and in-vivo studies, as well as clinical observations on silver-nylon, silver-intramedullary pins, silver oxide Foley catheters and silver-coated vascular prostheses have been performed during the past 30 years. Nevertheless, randomized clinical studies showing efficacy of such coated medical devices in high-risk patient populations are rare, have dealt with very small numbers of patients or are controversial. Physico-chemical, pharmacological and microbiological data explaining the antimicrobial efficacy of silver in prophylaxis of implants are presented here, as well as the scientific background for the established clinical benefits of silver-preparations in burns. PMID- 9868617 TI - The diagnosis of large joint sepsis. AB - The microbial aetiology of large joint sepsis is changing now that joint replacement therapy is becoming common place. The clinical history and examination may give important clues about the likelihood of infection and the possible involvement of unusual organisms. Newly introduced technology to improve the sensitivity of tests for the presence of micro-organisms in synovial fluid has not yet made a significant contribution to routine management. New imaging techniques such as magnetic resonance imaging are likely to improve patient management, but their utility is still under assessment. Arthroscopic biopsy to obtain material for culture and histology is particularly important in suspected chronic septic arthritis. Prosthetic joint infections present a particular challenge to microbiologists since the organisms involved are often found in small numbers and are common skin contaminants. Optimal microbiological management involves the taking of multiple (> or = 5) samples, careful processing to resuscitate organisms whilst avoiding contamination, and careful interpretation of results in the light of the clinical and histological picture. PMID- 9868618 TI - Early onset pneumonia in neurosurgical intensive care unit patients. AB - To investigate early onset pneumonia in a neurosurgical intensive care unit, we studied a cohort of patients over a 13-month period and compared neurotrauma (T) with non-neurotrauma (NT) patients. Data were abstracted from the infection surveillance database. Five hundred and sixty-five adults were hospitalized in the neurosurgical intensive care unit. 57.9% had trauma and 129 patients developed 152 episodes of pneumonia. Incidence rates, restricted to the 129 first episodes of pneumonia, were 20.1 versus 15.7/1000 patient days and 34.2 versus 27.9/1000 ventilation days, in the T and NT groups respectively. In both groups, the distribution of risk stratified by hospital days was bimodal, being highest during the first three days. However, the risk was higher for T patients (at day 3, 20/1000 ventilation days versus 10.2/1000 ventilation days). The daily risk peaked again at days 5 and 6, and thereafter remained low. Pneumonia occurring within the first three days, or early onset pneumonia (EOP), was associated with trauma (P = 0.036) and, in the NT group only, with a Glasgow coma scale score lower than 9 (P = 0.062). EOP was caused by Staphylococcus aureus (33%), Haemophilus spp. (23%), other Gram-positive cocci (22%), and other Gram-negative bacilli (GNB) (19%); whereas after the third day GNB other than Haemophilus spp. accounted for 45.4% of isolates (P = 0.11). This large series confirms the high incidence of EOP in neurosurgical intensive care units, particularly among trauma patients, in relation to risk factors different from those seen in other intensive care patients. Further studies are needed to elaborate specific preventive measures during early care. PMID- 9868619 TI - Assessment of person-to-person transmission of hantavirus pulmonary syndrome in a Chilean hospital setting. AB - Person-to-person transmission of Andes hantavirus among healthcare workers was reported in Argentina for the first time in 1996. To determine whether such transmission of the virus occurred during a 1997 outbreak of hantavirus pulmonary syndrome (HPS) in southern Chile due to Andes virus, we conducted a serological and epidemiological study in the Coyhaique Regional Hospital, where the majority of HPS patients were admitted. Workers in every department of the hospital were evaluated for immunoglobulin G (IgG) and IgM hantavirus antibodies using the enzyme-linked immunosorbent assay (ELISA). A standardized questionnaire was used to determine the type and extent of exposure to HPS patients, as well as other potential risk factors and previous febrile respiratory illnesses requiring hospitalization. Less than half (44%) reported always using gloves when touching patients or their secretions; respiratory protection was not used. Of the 319 participants (87.9% of those eligible), 12 (3.7%) had IgG antibodies. This finding is consistent with the antibody prevalence in the community in which the participants lived. Of the 12 positive healthcare workers, six reported contact with HPS patients. A similar exposure was found in those who tested negative [6/140 (4%) compared to 6/179 (3%), P = 0.66]. There was no significant difference in the types of hospital activities performed or the number of hospitalizations for febrile respiratory illnesses between antibody-positive and antibody-negative individuals. These data suggest that there was no person-to person transmission among healthcare workers during a recent outbreak of HPS in Southern Chile in 1997, despite the inconsistent use of any precautions against transmission. PMID- 9868620 TI - Hepatitis G virus infection from needle-stick injuries in hospital employees. AB - A new RNA virus, hepatitis G virus (HGV) is known to be transmissible by blood transfusion. The aim of this study was to assess whether HGV is an occupational risk to hospital employees as a result of exposure to needle-stick injuries. Among 220 cases of needle-stick injuries, 21 employees were contaminated with HGV. Initially none of the 21 recipients were HGV positive. Fourteen of the 21 recipients were followed up and further tested for HGV RNA and serum anti envelope (E2) specific antibody. None of the 21 recipients exposed to HGV developed liver function abnormalities, but one of the 14 recipients became positive for HGV RNA after the injury. Anti-E2 was negative in all recipients tested. These findings suggests a low clinical risk of occupational exposure to HGV in hospital employees. Nevertheless HGV is transmissible by needle-stick injury. PMID- 9868621 TI - Now wash your hands? A survey of hospital handwashing facilities. AB - We studied the adequacy of handwashing facilities on elderly-care wards in seven UK hospitals. We found 10.9% of sinks were inaccessible, 12% of sinks did not have any cleansing agent, 93.2% of ward sinks and 79% of treatment room sinks had no antiseptic agent. We conclude that in our survey, handwashing facilities were inadequate and have suggested a standard checklist for hospital sinks. PMID- 9868622 TI - An outbreak of extended-spectrum, beta-lactamase-producing Salmonella senftenberg in a burns ward. AB - A strain of Salmonella senftenberg resistant to ceftazidime, gentamicin, chloramphenicol and ciprofloxacin was isolated from burn wounds of eight patients on a burns ward of a hospital in Delhi, India. The organism, which had probably been spread from patient to patient on staff hands, produced the extended spectrum beta-lactamase SHV-5 and the aminoglycoside-modifying enzymes AAC(3)II + AAC(6'). The strain was not isolated from stool cultures of any of the patients or staff, apart from the index patient who had a history of diarrhoea and fever before admission. The outbreak ended in three weeks, after the implementation of strict handwashing. This is the first report of SHV-5 beta-lactamase in Salmonella spp. and also the first report of SHV-5 in India. The extended spectrum beta-lactamases that have been reported in Salmonella spp. now include the Group 2 be enzymes SHV-2, SHV-5, TEM-3, TEM-25, TEM-27, CTX-M2, PER-1 and PER 2, and the Group 1 enzymes DHA-1 and CMY-2. The types of extended-spectrum beta lactamases produced by salmonellas, their association with aminoglycoside resistance and their geographical distribution are now similar to those seen in klebsiella. Increasing antibiotic resistance in these organisms is reducing therapeutic options for the treatment of invasive disease. PMID- 9868623 TI - Cross-contamination potential of saliva ejectors used in dentistry. AB - It has been postulated that evacuation systems used in dentistry could be a source of cross-contamination between patients through backflow of bacteria dislodged from the saliva ejector tubings. The bacterial microflora associated with these systems was characterized using transmission electron microscopy (TEM) and microbiological cultures. The potential for backflow was investigated by a study of pressure differentials in evacuation system tubing and by the presence of bacteria in backflow samples. Evacuation lines were coated with microbial biofilms in which microcolonies of Gram-positive cocci and Gram-negative bacilli predominated, embedded in an extensive polysaccharide matrix. Most bacteria were metabolically active. Occasionally, buccal material such as collagen, fibrin and eukaryotic cell debris was observed. In other experiments, flow reversal was detected several times during saliva ejector use though each of these events was brief (less than 0.1 s). Aspiration of saliva, or occlusion of the mouthpiece opening by the oral mucosa, were the major factors leading to backflow episodes. Bacteria associated with backflow were found in almost 25% assays, with counts ranging from 1-300 cfu/occurrence. The majority of the bacteria isolated from biofilm or backflow samples were staphylococci, micrococci and non-fermentive Gram-negative rods. Pathogens such as Pseudomonas aeruginosa and Staphylococcus aureus were also isolated from backflow fluids. No oral streptococci could be recovered from biofilms in the tubing beyond 15 min from the last saliva ejector use however, suggesting that these species did not survive in the biofilms. These data suggest, although without direct proof of cross-contamination, the possible existence of an infectious risk associated with oral evacuation systems, as potential pathogens may be shed from tubing biofilms following backflow. Even if the risk of cross-contamination between patients is considered to be low, the necessity for regular disinfection of these systems must be stressed, since biofilms can serve as a reservoir for pathogens or harbor potentially infectious material. PMID- 9868624 TI - Antimicrobial efficacy of biocides tested on skin using an ex-vivo test. AB - An ex-vivo test was used to evaluate the activity of antimicrobials against three microorganisms, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The ex-vivo test is a carrier test using freshly excised animal skin samples maintained in viable conditions for a short period of time. Skin samples came from a veterinary practice and were excised from either dogs or cats. The antimicrobial activity of povidone iodine, chlorhexidine diacetate, cetrimide and benzalkonium chloride was also evaluated with suspension and glass-carrier tests. Generally, the activity of the antimicrobials tested was reduced when applied to the skin surface. Apart from povidone iodine (2%) against S. aureus, the biocides investigated failed to achieve a 5 log10 reduction in bacterial titre when tested with the ex-vivo method. There was no significant difference in reduction of bacterial titres after treatment with antimicrobials between the glass-carrier and the suspension tests. Furthermore, the drying process of bacterial inoculum was less detrimental on skin than on glass surfaces. This study confirmed that the activity of a biocide tested in suspension or on an inanimate surface did not reflect its activity when tested on skin. Further development of the ex-vivo test may be useful, especially for testing the antimicrobial activity of formulations with antiseptic properties. PMID- 9868625 TI - Viral haemorrhagic fever guidelines. PMID- 9868626 TI - Importance of air quality and related factors in the prevention of infection in orthopaedic implant surgery. PMID- 9868627 TI - Urine antigen detection can be quicker than PCR in the diagnosis of meningococcal disease. PMID- 9868628 TI - The association of methicillin resistant Staphylococcus aureus and pericarditis. PMID- 9868629 TI - Reflex responses of bladder motility after stimulation of interspinous tissues in the anesthetized rat. AB - OBJECTIVES: To determine the effects of somatic stimulation, including noxious chemical stimulation of interspinous tissues, on bladder motility in the anesthetized rat. METHODS: Changes in pressure in the previously quiescent bladder were measured in anesthetized adult female Wistar rats after various forms of noxious and innocuous somatic stimulation, including injection of the thoracic and lumbar interspinous tissues with capsaicin. Measurements were taken in both central nervous system-intact and spinalized animals, as well as in animals in whom the pelvic nerves had been transected bilaterally. Changes in bladder pressure were also measured in response to electrical stimulation of the primary dorsal ramus of lumbar spinal nerves. RESULTS: Noxious and innocuous stimulation of the fore- and hindpaws and the skin overlying the sacrum generally failed to elicit discernible changes in bladder pressure. However, capsaicin injection of thoracic and lumbar interspinous tissues produced profound and long lasting increases in bladder pressure. There were no significant differences in the responses to thoracic, as opposed to lumbar, stimulation. Spinalization above the level of stimulation abolished the response to capsaicin injection, as did bilateral transection of the pelvic nerves. CONCLUSION: In general, pressure in the quiescent bladder was relatively insensitive to somatic stimulation. However, noxious chemical stimulation of the interspinous tissues produced a nonsegmentally organized, supraspinal, parasympathetically mediated reflex increase in bladder tone. PMID- 9868630 TI - Postural changes of the cervical spine in patients with nontoxic goiter. AB - PURPOSE: To investigate differences in cervical spine posture and range of motion and self-reported neck pain and headache between patients with nontoxic goiter compared with a matched control group. DESIGN: An observational, controlled, blinded study. SETTING: The ambulatory outpatient facility of a university hospital. PARTICIPANTS: Twenty-five nontoxic goiter patients and 25 matched nongoiterous control subjects from the Department of Endocrinology. INTERVENTION: Participants were X-rayed from a lateral position in neutral, full flexion and full extension, and the radiographs were evaluated by a blinded examiner for anterior head carriage, maximal flexion, maximal extension and the extent and severity of any degenerative changes in the cervical spine. The degree of postural neck muscle tenderness was evaluated by a blinded rheumatologist using a validated Total Tenderness Score system. In addition, the two groups were compared for their self-reported frequency of neck pain and headaches. RESULTS: A significant increase in anterior head carriage was found among the goiter patients (p = .01), together with a corresponding decrease in flexion (p = .01), whereas the corresponding increase in extension was not statistically significant (p = .16). A higher prevalence of headaches was found in the goiter group (p = .06), but there was no difference in neck muscle tenderness (p = .40) or frequency of neck problems (p = .40) between the groups. The severity of degenerative changes in the cervical spine (p = .22) and the number of vertebral levels with degenerative changes (p = .13) were similar in the two groups. CONCLUSIONS: Goiters of > 100 g seem to alter the posture of the cervical spine, possibly resulting in a tendency for more frequent headaches. The changes do not seem to cause more neck pain, muscle tenderness or degeneration of the cervical spine. PMID- 9868631 TI - Number of sampling movements and manual stiffness judgments. AB - OBJECTIVE: It has been suggested that when using posteroanterior pressure to assess the stiffness of the spine, no more than two or three oscillations should be used. This study sought to examine this clinical impression by investigating the effect of the number of sampling movements on stiffness perception. DESIGN: Perceptual study using a mechanical device to provide stiffness stimuli, with university staff and students as judges. SETTING: University psychophysics laboratory. INTERVENTIONS: In part 1, subjects were directed to sample the stimuli a prescribed number of times (either 1, 2, 3, 4 or 5 pushes), whereas in part 2, subjects were free to choose the number of sampling movements. MAIN OUTCOME MEASURE: Measures of interstimulus discriminability and bias. RESULTS: An inverted-U relationship between the number of directed sampling movements and stiffness discriminability was found, with three cycles providing best discriminability. When subjects were given a choice, most chose to use three cycles. The number of sampling movements had no effect on bias. CONCLUSION: This study confirms the hypothesis under investigation and suggests that therapists will be maximally sensitive when using three testing cycles of posteroanterior pressure to assess stiffness. PMID- 9868632 TI - The effect of upper cervical or sacroiliac manipulation on hip flexion range of motion. AB - OBJECTIVES: To compare the effectiveness of an upper cervical manipulation and a manipulation of the sacroiliac joint for increasing hip range of motion. DESIGN: Clinical cohort study. SETTING: Macquarie University Centre for Chiropractic Outpatient Clinic. SAMPLE: Fifty-two randomly chosen university students aged 18 to 34 yr. METHOD: A reliable hand-held dynamometer was used to determine the end point of range of motion before and after the application of a treatment. Three groups of subjects were created: cervical manipulation, sacroiliac manipulation and sham/placebo. Range of motion of the hip in flexion (SLR) was used as the independent variable. RESULTS: The two manipulative treatments resulted in increased flexion range of motion at the hip. Statistical analysis revealed that only the upper cervical manipulation procedure increased hip flexion range of motion significantly. CONCLUSION: The results suggest that manual therapy of the neck may affect hip range of motion in normal adults. Findings such as these may indicate the existence of a link between the cervical spine and the lower extremity. PMID- 9868633 TI - Diagnostic value on signs of subluxation of cervical vertebrae with radiological examination. AB - OBJECTIVE: To investigate the relationship between radiographic signs of subluxation in the cervical vertebrae and their clinical diagnostic value. DESIGN: Controlled, clinical study. SETTING: Institute of Clinical Anatomy and Biomechanics and NanFang Hospital of the First Military Medical University, Guangzhou, China. SUBJECTS: Eighty-seven subjects with cervicodynia and 21 asymptomatic volunteers. INTERVENTIONS: Radiological signs of subluxation from anteroposterior, lateral, open-mouth and dynamic radiographs of the cervical vertebrae of the subjects were measured and analyzed. MAIN OUTCOME MEASURES: The right and left odontoid lateral mass interspace, divergence of the spinous processes, sign of double contour and position of odontoid process were studied. RESULTS: The bilateral odontoid lateral mass interspaces were asymmetrical in most cases, and the divergence of spinous processes, sign of double contour and position of odontoid process were also common. Cervical vertebrae C5, C4 and C6 showed no special variations. CONCLUSION: There was little evidence to support the contention that signs of subluxation in the cervical vertebrae are of diagnostic significance. Subluxation should be defined in two ways: as a purely roentgenological diagnosis and as a combination of roentgenological signs with clinical signs. PMID- 9868634 TI - Influence of a Biokinergia session on cardiorespiratory and metabolic adaptations of trained subjects. AB - BACKGROUND AND PURPOSE: Biokinergia (BK) is a massage aimed at improving "biological" potential. An initial pilot study demonstrated that a BK session could alter blood lactate concentration during prolonged exercise. OBJECTIVE: To confirm the effect of BK on performance and physiological adaptations during maximal aerobic muscular exercise. METHODS: Two groups of 10 subjects each performed a maximal progressive exercise on a cycle ergometer before and 10 days after a BK session (B group) or a feigned one (C group) in a simple-blinded manner. RESULTS: No modifications were noticed in the C group. After BK in the B group, maximum oxygen consumption was slightly increased (7%) and submaximal heart rate slightly lowered (4.7%) in 65% of the subjects; tidal volume was higher and breathing frequency was lower for 45% of the subjects. CONCLUSION AND DISCUSSION: The slight modifications induced by BK suggest that BK altered the sympathetic nervous system, but this needs confirmation. PMID- 9868635 TI - Rear-end impacts: vehicle and occupant response. AB - BACKGROUND: There is a controversy regarding the likelihood of injuries sustained when one car strikes another at a relatively low speed with little or no vehicle damage. Plaintiffs often claim injuries whereas defendants counterclaim that injuries could not have occurred with such a relatively minor impact. OBJECTIVE: To review the dynamics of low-speed rear-end collisions resulting in little or no visible damage and to decide whether occupant injury can occur; also, to discuss diagnostic examination and treatment that may be helpful to the clinical practitioner. DATA SELECTION: A Medline search for articles discussing low-speed rear-end collisions was conducted. Other articles and studies were reviewed that discussed low-speed rear-end collisions and factors impacting the neuromusculoskeletal system relevant to clinical practitioners. Articles included were human low-speed rear-end tests, lab tests on cadavers, automotive engineering articles, and peer-reviewed journal articles on whiplash. A few live animal and simulation studies were considered for the background of possible injury mechanism and vehicular deformation. Excluded were non-rear-end collison and single case reports. DATA SYNTHESIS: The data were studied to find a relationship between the resultant vehicle dynamics and occupant movement, biological mechanisms of injury and the neurological mechanisms causing complaints. Data were also studied to investigate objective findings supporting subjective complaints. CONCLUSION: In low-impact collisions, there are usually no skid marks and minor or no visible damage to the vehicle. There is a lack of relationship between occupant injury, vehicle speed and/or damage. There does not seem to be an absolute speed or amount of damage a vehicle sustains for a person to experience injury. Crash tests indicate that a change of vehicle velocity of 4 km/hr (2.5 mph) may produce occupant symptoms. Vehicle damage may not occur until 14-15 km/hr (8.7 mph). Occupant soft tissue and joint injuries resulting from low speed vehicle collisions respond positively to afferent stimulation of mechanoreceptors. The diagnosis of the occupant injuries relies on standard orthopedic neurological testing, autonomic concomitant signs and qualitative and quantitative testing. PMID- 9868636 TI - Chondrosarcoma and myositis ossificans. AB - OBJECTIVE: To discuss the clinical radiographic findings in a 70-yr-old woman suffering from chondrosarcoma. CLINICAL FEATURES: The patient experienced right SI pain present initially only at night. She later developed morning numbness. An X-ray examination revealed a flocculent calcification in the right buttock region. Computed tomographic scans confirmed the diagnosis. INTERVENTION AND OUTCOME: Initial palliative care continued until surgery was performed to resect the area. CONCLUSION: Chondrosarcoma is a severe disease that must be differentiated from myositis ossificans. PMID- 9868637 TI - Torque misuse revisited. PMID- 9868638 TI - The effects of X-axis vertebral translation on projected Y-axis vertebral rotation. PMID- 9868639 TI - Reliability of spinal displacement analysis on plain X-rays: a review of commonly accepted facts and fallacies with implications for chiropractic education and technique. PMID- 9868640 TI - Rocky mountain spotted fever. PMID- 9868641 TI - Photo quiz. Infection due to Dracunculus medinensis. PMID- 9868642 TI - Infection due to Yersinia enterocolitica in a series of patients with beta thalassemia: incidence and predisposing factors. AB - Over 15 years, 14 patients with yersiniosis in two North American comprehensive thalassemia clinics (0.6 cases per 100 patient-years) presented with fever (100%), diarrhea (86%), right-lower-quadrant abdominal pain (71%), bacteremia (57%), a palpable abdominal mass (36%), and pharyngitis (28%). Clinically apparent infection occurred within 10 days of blood transfusion in 57% of patients. Nine patients (64%) had only a modest elevation in serum level of ferritin (< 2,000 micrograms/L). Patients with focal abdominal findings had a higher body iron burden, as estimated by the serum ferritin level, and significant intraabdominal suppurative complications. Two patients were not receiving iron-chelating therapy with deferoxamine; one patient was receiving the experimental chelator deferiprone (L1). Iron-loaded patients with beta thalassemia are at greatly increased risk for severe yersiniosis, even when their body iron burden (as indicated by the serum ferritin level) is only moderately elevated and they are not receiving iron-chelating therapy with deferoxamine. PMID- 9868643 TI - Iron and infection. PMID- 9868644 TI - Prophylaxis with weekly versus daily fluconazole for fungal infections in patients with AIDS. AB - We compared the efficacy of a 400-mg once-weekly dosage versus a 200-mg daily dosage of fluconazole for the prevention of deep fungal infections in a multicenter, randomized, double-blind trial of 636 human immunodeficiency virus infected patients to determine if a less intensive fluconazole regimen could prevent these serious but relatively infrequent complications of AIDS. In the intent-to-treat analysis, a deep fungal infection developed in 17 subjects (5.5%) randomly assigned to daily fluconazole treatment and in 24 (7.7%) given weekly fluconazole during 74 weeks of follow-up (risk difference, 2.2%; 95% confidence interval [CI], -1.7% to 6.1%). Thrush occurred twice as frequently in the weekly versus daily fluconazole recipients (hazard ratio, 0.59; 95% CI, 0.40-0.89), and in a subset of patients evaluated, fluconazole resistance was infrequent. Fluconazole administered once weekly is effective in reducing deep fungal infections in patients with AIDS, but this dosage is less effective than the 200 mg-daily dosage in preventing thrush. PMID- 9868645 TI - Antifungal prophylaxis with weekly fluconazole for patients with AIDS. PMID- 9868646 TI - The incidence and spectrum of AIDS-defining illnesses in persons treated with antiretroviral drugs. AB - The incidence and spectrum of primary AIDS-defining illnesses in human immunodeficiency virus-positive patients receiving antiretroviral drugs may have changed since the introduction of newer antiretroviral agents. We performed a retrospective analysis of patients enrolled in the British Columbia Drug Treatment Program who were ever prescribed antiretroviral drugs between 1 January 1994 and 31 December 1996. Rates were calculated on a 6-month basis. There were 344 AIDS cases diagnosed among 2,533 participants between 1994 and 1996. The incidence of primary AIDS diseases decreased from 1994 to 1996, with a sharp decline in 1995 and 1996. There was no statistically significant change in the incidence of primary AIDS diagnoses relative to one another, and Pneumocystis carinii pneumonia and Kaposi's sarcoma remain the most common AIDS index diagnoses. In patients receiving antiretroviral therapy in the modern era, the incidence of AIDS-defining illnesses has decreased substantially, but the spectrum of AIDS-defining illnesses remains unchanged. PMID- 9868647 TI - What is the relevance of antiretroviral therapy that does not include protease inhibitors? PMID- 9868648 TI - Successful medical therapy for deeply invasive facial infection due to Pythium insidiosum in a child. AB - Pythiosis occurs in animals and humans who encounter aquatic habitats that harbor Pythium insidiosum. Drug therapy for deeply invasive infections with this organism has been ineffective in humans and animals; patients have been cured only by radical surgical debridement. A 2-year-old boy developed periorbital cellulitis unresponsive to antibiotic and antifungal therapy. The cellulitis extended to the nasopharynx, compromising the airway and necessitating a gastrostomy for feeding. P. insidiosum was isolated from surgical biopsy specimens of the affected tissue. On the basis of in vitro susceptibility studies of the isolate, the patient was treated with a combination of terbinafine and itraconazole. The infection resolved over a period of a few months. The patient remained well 1.5 years after completing a 1-year course of therapy. Cure of deep P. insidiosum infection is feasible with drug therapy. PMID- 9868649 TI - Use of an immunotherapeutic vaccine to treat a life-threatening human arteritic infection caused by Pythium insidiosum. AB - A 14-year-old Thai boy presented because of a history of headache, mandibular swelling, and facial nerve palsy. A microorganism identified as Pythium insidiosum was cultured from the mandibular abscesses. Despite treatment with amphotericin B, iodides, ketoconazole, and surgery, the infection progressed. Magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) of the neck revealed an aneurysm in the external carotid artery. The aneurysm was removed. MRA performed later showed stenosis of the internal carotid artery. Immunotherapy was recommended as a last resort. One hundred microliters of the P. insidiosum vaccine was subcutaneously injected into the patient's left shoulder, and 14 days later a similar dose was administered. Four weeks following the first vaccination, the patient's headache had disappeared, the facial swellings had dramatically diminished, the cervical lymph node had shrunk, and the proximal left internal carotid artery stenosis had significantly improved. One year after the vaccinations, the boy was considered clinically cured. PMID- 9868650 TI - Does in vitro susceptibility to rifabutin and ethambutol predict the response to treatment of Mycobacterium avium complex bacteremia with rifabutin, ethambutol, and clarithromycin? Canadian HIV Trials Network Protocol 010 Study Group. AB - The in vitro susceptibilities of baseline Mycobacterium avium complex (MAC) blood isolates from 86 patients with AIDS who were treated with clarithromycin, ethambutol, and rifabutin were determined to examine whether these results predict bacteriologic response to treatment. No patient received prior prophylaxis with clarithromycin or azithromycin. Minimum inhibitory concentrations (MICs) of clarithromycin for all isolates were < or = 2 micrograms/mL. The median MIC of rifabutin was between 0.25 and 0.5 microgram/mL, and all isolates were susceptible to < or = 2 micrograms of rifabutin/mL. The median MIC of ethambutol was 4 micrograms/mL, and the MIC90 was 8 micrograms/mL. There was no correlation between ethambutol susceptibility and subsequent bacteriologic clearance. At all time points through week 12, bacteriologic clearance occurred more frequently in patients with isolates for which MICs of rifabutin were lower, but this difference was statistically significant only at week 2. Susceptibility testing for baseline MAC isolates from AIDS patients not previously treated with clarithromycin or azithromycin does not appear to be useful in guiding therapy. PMID- 9868652 TI - Randomized trial of lipid-based amphotericin B for invasive aspergillosis in neutropenic hosts is an important step forward. PMID- 9868651 TI - An EORTC international multicenter randomized trial (EORTC number 19923) comparing two dosages of liposomal amphotericin B for treatment of invasive aspergillosis. AB - This is the first completed prospective randomized clinical efficacy trial of antifungals in the treatment of invasive aspergillosis (IA) and the first to compare the clinical efficacy of two dosages of liposomal amphotericin B (L-AmB) for IA in neutropenic patients with cancer or those undergoing bone marrow transplantation. Eighty-seven of 120 patients were eligible and evaluable. Clinical responses were documented for 26 (64%) of 41 patients receiving 1 mg/(kg.d) (L-AmB-1) and 22 (48%) of 46 receiving 4 mg/(kg.d) (L-AmB-4). Radiologic response rates were similar: 24 (58%) of the L-AmB-1 recipients and 24(52%) of the L-AmB-4 recipients. The six-month survival rates were 43% (L-AmB 1) and 37% (L-AmB-4). These differences were not significant. The numbers of deaths directly due to IA at 6 months were similar: 9 (22%) of 41 L-AmB-1 recipients and 9 (20%) of 46 L-AmB-4 recipients. No other variable independently influenced survival, apart from central nervous system IA. L-AmB is effective in treating approximately 50%-60% of patients who have IA. A 1-mg/(kg.d) dosage is as effective as a 4-mg/(kg.d) dosage, and no advantages to use of the higher, more expensive, dosage has been observed. PMID- 9868653 TI - Medicare coverage of outpatient ambulatory intravenous antibiotic therapy: a program that pays for itself. AB - A number of studies have documented the safety, efficacy, and cost-effectiveness of outpatient intravenous (i.v.) antibiotic therapy for patients with infectious diseases. Nevertheless, Medicare policy prohibiting coverage of outpatient, self administered drugs has severely limited access of Medicare patients to ambulatory i.v. therapy, thus forcing them to rely on more costly, impatient hospital care. To test the hypothesis that a new Medicare benefit providing coverage for ambulatory i.v. antibiotic therapy could significantly reduce the program's expenditures for the treatment of infectious diseases (including pneumonia, osteomyelitis, cellulitis, and endocarditis), a cost model was constructed with use of patient care information from the clinical literature as well as clinical experts, Medicare data, and other medical claims databases. The model shows cumulative 5-year savings of nearly $1.5 billion associated with the new Medicare benefit. Policy makers should consider implementing such a benefit. PMID- 9868654 TI - Medicare and outpatient therapy for infectious diseases. PMID- 9868655 TI - Fatal pancarditis associated with human granulocytic Ehrlichiosis in a 44-year old man. AB - Human cases of infection with a granulocytotropic Ehrlichia species closely related to Ehrlichia equi are now being described with increasing frequency in the United States, especially in areas where Lyme disease is already endemic. We describe a case of fatal pancarditis during the course of human granulocytic ehrlichiosis (HGE) in a 44-year-old outdoor worker who was previously treated for presumptive Lyme disease. Serological and molecular diagnostic tests for Borrelia burgdorferi and Babesia microti infections were negative. Postmortem serum specimens were seroreactive for HGE, and molecular evidence of infection with the HGE agent was obtained. These findings suggest that carditis may be a manifestation of HGE, further complicating the differential diagnosis of tick borne illness. PMID- 9868656 TI - Epidemiological and clinical aspects of invasive group A streptococcal infections and the streptococcal toxic shock syndrome. AB - In a retrospective study of invasive infections due to group A Streptococcus (GAS) in Stockholm during 1987 to 1995, the average incidence per 100,000 residents per year was 2.3, varying between 3.7 per 100,000 (in 1988) and 1.3 per 100,000 (in 1993). Incidence was 1.8 in the age group of 0-4 years but otherwise increased by age, from 0.48 in the age group of 5-14 years to 6.1 among those over 65 years of age. A review of 151 invasive episodes occurring in 1983-1995 showed cyclic increases of infections due to T1M1-serotype strains during 1986 1990 and 1993-1995. The T1M1 serotype accounted for 27 (20%) of 135 available GAS strains. Streptococcal toxic shock syndrome (STSS) developed in 19 (13%) of the 151 episodes. The case fatality rate was 11% overall but 47% among patients with STSS. In a multivariate logistic regression model, STSS was associated with a history of alcohol abuse (odds ratio [OR], 6.3; P = .004) and infection with a T1M1 strain (OR, 6.7; P = .007). Case fatality was associated with age (OR, 14.5; P = .08), immunosuppression (OR, 4.7; P = .02), and STSS (OR, 21.5; P < .0001) but not with T1M1 infection. Hypotension was significantly associated with a fatal outcome, regardless of whether STSS developed (P < .0001). PMID- 9868657 TI - Invasive pulmonary infection due to Scedosporium apiospermum in two children with chronic granulomatous disease. AB - Scedosporium apiospermum is an opportunistic fungus in humans. The incidence of S. apiospermum infection in patients with acquired neutropenia (e.g., patients receiving chemotherapy and bone marrow transplant recipients) is steadily increasing. S. apiospermum has poor in vitro susceptibility to "conventional" antifungal agents, rendering the management of infections complex. Patients with chronic granulomatous disease (CGD) are highly susceptible to fungal infections, which are mostly due to Aspergillus species. We describe two children with CGD and invasive pulmonary infection due to S. apiospermum. Both patients were treated with antifungal therapy including azole derivatives (itraconazole or voriconazole) and surgical resection of infected tissues. These cases highlight that scedosporium infection can closely mimic aspergillus infection and should be considered in any case in which there is a failure to respond to appropriate "conventional" antifungal therapy. We also suggest that the emergence of this pathogen may have been favored by long-term use of amphotericin B in both patients. PMID- 9868658 TI - Serotype distribution and prevalence of resistance to benzylpenicillin in three representative populations of Streptococcus pneumoniae isolates from the coast of Kenya. AB - As surveillance data from sub-Saharan Africa are few, three representative populations of Streptococcus pneumoniae isolates were examined in Kenya for serotype distribution and Etest minimum inhibitory concentrations (MICs) of benzylpenicillin: (1) 75 lung aspirate or blood culture isolates from 301 consecutive adult patients with pneumonia, (2) 112 invasive isolates from continuous pediatric inpatient surveillance over 4 years, and (3) 97 nasopharyngeal isolates from systematically selected sick children. The proportions with benzylpenicillin MICs of > or = 0.1 microgram/mL were 0.27, 0.29, and 0.47, respectively. Vaccine-related serotypes accounted for 96% of invasive isolates from children and 90% of those from human immunodeficiency virus (HIV)-seropositive adults. Serotype 1 accounted for 44% of pneumococci from HIV-seronegative patients but only 5% of those from HIV-seropositive patients (P = .0002). Of serotype 1 isolates, 98% were susceptible to benzylpenicillin, but serogroups 13, 14, 19, and 23 were strongly associated with an MIC of > or = 0.1 microgram/mL. PMID- 9868659 TI - Are the Duke criteria superior to the Beth Israel criteria for the diagnosis of infective endocarditis in children? AB - Accurate diagnosis of infective endocarditis may be difficult. The Beth Israel criteria and the newer Duke criteria assign probability to the diagnosis of infective endocarditis on the basis of the presence of common features and manifestations. We reviewed 111 cases of pediatric infective endocarditis diagnosed and treated over 19 years. Each case was classified by the two criteria, and the results were compared. Of 111 cases, 73 (66%) and 18 (16%) were classified as definite by the Duke criteria and the Beth Israel criteria, respectively. No cases were rejected by the Duke criteria, while 21 (19%) of 111 were rejected by the Beth Israel criteria. In 18 pathologically proven cases, reanalysis without pathological data showed that the Duke criteria had significantly greater sensitivity (83%) than the Beth Israel criteria (67%) (P < .03). Echocardiographic evidence was required in 22 cases for definite classification by the Duke criteria; none were rejected, however, when echocardiographic findings were ignored. Our results suggest that the Duke criteria are superior to the Beth Israel criteria for the diagnosis of pediatric infective endocarditis. PMID- 9868660 TI - Safety and efficacy of intravenous sodium stibogluconate in the treatment of leishmaniasis: recent U.S. military experience. AB - The efficacy and toxicity of sodium stibogluconate (SSG) at a dosage of 20 mg/(kg.d) for either 20 days (for cutaneous disease) or 28 days (for visceral, mucosal, or viscerotropic disease) in the treatment of leishmaniasis is reported. Ninety-six U.S. Department of Defense health care beneficiaries with parasitologically confirmed leishmaniasis were prospectively followed for 1 year. One patient was infected with human immunodeficiency virus; otherwise, comorbidity was absent. Clinical cure occurred in 91% of 83 cases of cutaneous disease and 93% of 13 cases of visceral/viscerotropic disease. Adverse effects were common and necessitated interruption of treatment in 28% of cases, but they were generally reversible. These included arthralgias and myalgias (58%), pancreatitis (97%), transaminitis (67%), headache (22%), hematologic suppression (44%), and rash (9%). No subsequent mucosal leishmaniasis was identified, and there were no deaths attributable to SSG or leishmaniasis. PMID- 9868661 TI - Tropical rat mite dermatitis: case report and review. AB - Six medical students inhabiting a centuries-old, rat-infested house in Lubeck, in northern Germany, were suffering from itching papules and seropapules. Prior to these patients' visit to our institute, their conditions had been diagnosed as pediculosis, scabies, or pulicosis and treated unsuccessfully with the antiparasitic agent lindane (0.3%). The final diagnosis, tropical rat mite dermatitis, was based on the identification of the arthropod Ornithonyssus bacoti, which has an unsegmented body with eight legs. No treatment was recommended, and the dermatitis disappeared within 2 weeks. Measures taken to prevent reinfestation included extermination of the rats and treatment of the rooms of the house with the acaricide benzyl benzoate. Because the mite O. bacoti spends a relatively short time on a host and penetrates the skin for feeding only, the application of an antiparasitic agent is not necessary. If indicated, treatment should be symptomatic. PMID- 9868662 TI - Ceftriaxone once daily for four weeks compared with ceftriaxone plus gentamicin once daily for two weeks for treatment of endocarditis due to penicillin susceptible streptococci. Endocarditis Treatment Consortium Group. AB - This randomized, multicenter, open-label study compared the efficacy and safety of monotherapy with 2 g of intravenous ceftriaxone once daily for 4 weeks with those of combination therapy with 2 g of intravenous ceftriaxone and 3 mg of intravenous gentamicin/kg once daily for 2 weeks as therapy for endocarditis due to penicillin-susceptible streptococci. Sixty-one patients were enrolled in the study. Clinical cure was observed for 51 evaluable patients both at termination of therapy and at the 3-month follow-up: 25 (96.2%) of 26 monotherapy recipients and 24 (96%) of 25 combination therapy recipients. Of the 23 patients in each treatment group who were microbiologically evaluable, 22 (95.7%) in each group were considered cured. No patient had evidence of relapse. Fourteen patients (27.5%) required cardiac surgery after initiation of treatment, including five monotherapy recipients and nine combination therapy recipients. Adverse effects were minimal in both treatment groups. We conclude that 2 g of ceftriaxone once daily for 4 weeks and 2 g of ceftriaxone in combination with 3 mg of gentamicin/kg once daily for 2 weeks are both effective and safe for the treatment of streptococcal endocarditis. PMID- 9868663 TI - Detecting serum antibodies to a purified recombinant proline-rich antigen of Coccidioides immitis in patients with coccidioidomycosis. AB - In previous work, antibodies in serum samples from patients with coccidioidomycosis were found to react with a proline-rich antigen (PRA) isolated from spherules of Coccidioides immitis, and the gene encoding this antigen was cloned. We expressed and purified recombinant PRA (rPRA) by removing the majority of amino acids contributed by the vector from the fusion protein. Purified rPRA reacted with serum IgG antibodies in 37 of 42 patients with culture-proven progressive pulmonary or extrapulmonary coccidioidal disease; specific antibodies in dilutions ranging from 1:40 to 1:102,400 were demonstrated (sensitivity, 88%). In contrast, for > 95% of patients without coccidioidomycosis reactivity of < 1:40 was demonstrated (specificity, 97%). Of 18 patients with primary self limited coccidioidomycosis, none had detectable antibodies in serum samples collected up to 141 days after illness began. The association of antibodies to rPRA with progressive infection may have prognostic value. PMID- 9868664 TI - Epidemiology of legionella pneumonia and factors associated with legionella related mortality at a tertiary care center. AB - Legionella pneumophila is an important pathogen that may cause nosocomial and community-acquired pneumonia in patients with normal or altered immunity. The epidemiology of 40 cases of legionella pneumonia in patients hospitalized between 1986 and 1994 was studied. Fourteen patients (35%) were solid organ transplant recipients. The calculated annual incidence of L. pneumophila infection was highest among lung transplant recipients (2.07 cases per 1,000 transplant-years). There was a trend toward reduced mortality rates and less severe disease among transplant patients vs. nontransplant patients: mortality rate, 36% vs. 54%; incidence of intubation, 50% vs. 69%; rate of concurrent infections, 29% vs. 38%; and overall rate of complications, 86% vs. 96%; respectively. In a multivariate analysis, factors independently associated with an increased mortality rate were nosocomial acquisition, need for intubation, formation of lung abscess or cavitation, and presence of pleural effusion. Thus, despite differing host immune responses, the most important prognostic factors affecting the outcome of legionellosis are nosocomial acquisition and the development of pulmonary complications. PMID- 9868665 TI - IgG responses to protein-conjugated pneumococcal capsular polysaccharides in persons who are genetically incapable of responding to unconjugated polysaccharides. AB - We have previously shown that the capacity to make IgG to pneumococcal capsular polysaccharides (PCPs) is inherited as an autosomal, mixed codominant trait. The purpose of this study was to determine whether this genetically determined unresponsiveness could be overcome by injection of protein-conjugated pneumococcal vaccines. Seven healthy adults who had failed to produce IgG to five or more of 10 representative PCPs after receiving pneumococcal vaccine and whose parents, siblings, and/or offspring had a similar lack of responsiveness received a series of protein-conjugated polysaccharide vaccines. Excellent IgG responses to most of the PCPs tested were eventually observed in five of the seven subjects after they received octavalent diphtheria toxoid-conjugated vaccine. Administration of certain protein-conjugated PCPs leads to IgG responses in some persons who lack the capacity to respond to unconjugated PCPs. PMID- 9868666 TI - Seroprevalence of human granulocytic ehrlichiosis among permanent residents of northwestern Wisconsin. AB - Four-hundred seventy-five permanent residents of Wisconsin were tested for antibodies to the agent of human granulocytic ehrlichiosis (HGE) by indirect immunofluorescent antibody (IFA) testing with Ehrlichia equi as antigen marker. Each resident completed a standard survey questionnaire about outdoor activities, animal and tick exposure, and any febrile illness during the preceding 12 months. Seventy-one serum samples (14.9%) contained E. equi antibodies. The mean IFA titer for seropositive residents was 250 (range, 80-10,240). Seropositive residents were older than seronegative ones (62 vs. 56 years; P = .019). None of the seropositive residents had a history suggestive of ehrlichiosis. There was no association between the IFA test outcome and specific demographic variables or history of tick bites. HGE appears to be a common subclinical or mild infection among residents in northwestern Wisconsin. PMID- 9868667 TI - Interleukin-10: potential benefits and possible risks in clinical infectious diseases. AB - Human interleukin-10 (IL-10) is a potent anti-inflammatory cytokine that inhibits the synthesis of the major proinflammatory cytokines and chemokines. IL-10 is the principal TH2-type cytokine that upregulates humoral immune responses and attenuates cell-mediated immune reactions. This cytokine has a number of immunomodulatory properties that might be clinically useful in a variety of inflammatory and infectious disease states. Clinical trials with human recombinant IL-10 are already in progress. Carefully selected patients with inflammatory conditions may benefit from IL-10 therapy if concomitant infectious diseases are recognized and treated appropriately. PMID- 9868669 TI - Potent antiretroviral therapy does more than just decrease viral load. PMID- 9868668 TI - High incidence of herpes zoster in patients with AIDS soon after therapy with protease inhibitors. AB - A high incidence of herpes zoster was noticed among patients with AIDS, shortly after addition of a protease inhibitor to their baseline treatment with nucleoside analogue reverse-transcriptase inhibitors. Within a median follow-up of 64 weeks (range, 34-103 weeks), 14 patients (7%) had a first episode or a recurrence of herpes zoster (6.2 episodes per 100 patient-years). No episodes of zoster were diagnosed before week 4. Twelve episodes (86%) occurred between weeks 4 and 16. The risk of zoster was independent of age, sex, type of protease inhibitor, and CD4+ lymphocyte count and viral load at baseline and month 1. A CD8+ lymphocyte proportion at baseline of > 66% (hazard ratio [HR], 10.6; 95% confidence interval [CI], 3.4-33.1) and an increase in CD8+ lymphocyte proportion at month 1 of > 5% (HR, 32; 95% CI, 8.1-126.4) were independently associated with the risk of herpes zoster. These data might be clinically useful for determining transient prophylaxis for those patients at high risk. PMID- 9868670 TI - Cellular immune responses to mycobacteria in healthy and human immunodeficiency virus-positive subjects in the United States after a five-dose schedule of Mycobacterium vaccae vaccine. AB - The safety and immunogenicity of heat-killed Mycobacterium vaccae vaccine were investigated in a pilot study assessing the feasibility of immunization to prevent mycobacterial disease in patients with human immunodeficiency virus (HIV) infection. Fifteen (seven healthy and eight HIV-positive subjects) received five doses of M. vaccae vaccine. Lymphocyte proliferation assays (LPAs) were performed using Mycobacterium avium sensitin (MAS) and M. vaccae sonicate (MVS). Vaccine was well tolerated in all 15 subjects with minimal induration at the vaccine site. LPAs for four of seven healthy vaccines were positive for MAS after immunization. Median responses to MAS and MVS that were determined by LPAs were consistently higher for the eight HIV-positive vaccinees than for the seven healthy controls. A five-dose series of M. vaccae vaccine is safe for both healthy and HIV-positive subjects and deserves further evaluation as a vaccine to prevent HIV-associated mycobacterial disease. PMID- 9868671 TI - Dual microsporidial infection due to Vittaforma corneae and Encephalitozoon hellem in a patient with AIDS. AB - A 46-year-old human immunodeficiency virus-infected Swiss citizen living in Tanzania presented with respiratory, abdominal, and urogenital complaints. Microsporidial spores were isolated from urine and a sinunasal aspirate and were propagated in MRC-5 cell cultures. Western blot analysis and riboprinting identified the sinunasal isolate as Encephalitozoon hellem. Electron microscopic investigation of the urine isolate revealed spores with diplokaryotic nuclei and five to six isofilar coils of the polar tube and sporonts with two or three diplokarya. All stages were enveloped by two membranes, corresponding to a cisterna of host endoplasmic reticulum studded with ribosomes. These characteristics have been described for the genus Vittaforma. Western blot analysis of this isolate revealed a banding pattern identical to that of the Vittaforma corneae reference isolate. Part of the small subunit rRNA gene was amplified, sequenced (239 base pairs), and found to be identical to that of V. corneae. This is the second isolation of V. corneae and the first description of urinary tract infection due to V. corneae in a patient with AIDS. PMID- 9868672 TI - Acyclovir-resistant herpes zoster in human immunodeficiency virus-infected patients: results of foscarnet therapy. AB - We retrospectively studied 18 consecutive cases of acyclovir-resistant zoster. All the patients had chronic skin lesions that failed to heal despite treatment with intravenous acyclovir (30 mg/[kg.d]) in 15 cases and oral acyclovir (4 g/d) in three cases for > 10 days. The mean CD4+ cell count was 20 x 10(6)/L. The mean number of previous zoster episodes was 1.53. Fifteen of the 16 patients evaluable for previous acyclovir treatment had received the drug. Thirteen patients were treated with intravenous foscarnet (200 mg/[kg.d]) for a mean of 17.8 days. Complete healing was observed in 10 (77%) of the 13 treated patients. Zoster relapsed after cessation of foscarnet therapy in five of the 10 responding patients. The median time to relapse was 110 days. Four patients died of varicella-zoster virus-associated visceral complications. These results show that acyclovir-resistant zoster has a poor prognosis but responds well to foscarnet therapy. PMID- 9868673 TI - Coccidioidomycosis in Arizona: increase in incidence from 1990 to 1995. AB - The number of cases of coccidioidomycosis (incidence) reported to the Arizona Department of Health Services increased from 255 (7.0 per 100,000 population) in 1990 to 623 (14.9 per 100,000 population) in 1995 (P < .001). Four counties in the south central region of the state, which contained 80% of the state's population, had the largest increase and accounted for 95% of all cases in 1995. Cases in persons aged 65 years or older and men were reported more frequently (for both, P < .001). During 1995, 890 patients were discharged from Arizona hospitals with a diagnosis of coccidioidomycosis. Rates of hospitalization were greater among persons aged 55 years or older, men, and African-American (for all three, P < .01). Of the hospitalized patients, 48 died, and 12 (25%) of these patients had a concurrent diagnosis of human immunodeficiency virus infection. These data demonstrate that coccidioidomycosis is a growing health problem in Arizona. PMID- 9868674 TI - Genital infection due to Mycobacterium genavense in a patient with AIDS. PMID- 9868675 TI - Septic synovitis and arthritis due to Corynebacterium striatum following an accidental scalpel injury. PMID- 9868676 TI - Detection of specific cellular immune response to Bartonella henselae in a patient with cat scratch disease. PMID- 9868677 TI - Mycoplasma pneumoniae and second-degree heart block. PMID- 9868678 TI - Persistent nephrolithiasis after discontinuation of indinavir therapy. PMID- 9868679 TI - Acute human immunodeficiency virus infection with severe respiratory and renal failure. PMID- 9868680 TI - Azithromycin therapy for scrub typhus during pregnancy. PMID- 9868681 TI - Gynecomastia associated with indinavir therapy. PMID- 9868682 TI - Clostridium difficile diarrhea after use of tacrolimus following renal transplantation. PMID- 9868683 TI - Pacemaker-induced endocarditis due to Propionibacterium acnes. PMID- 9868684 TI - Isolated cavitary pulmonary Mycobacterium avium complex infection in a patient with AIDS. PMID- 9868685 TI - Pulmonary tuberculosis due to Mycobacterium microti in a human immunodeficiency virus-infected patient. PMID- 9868686 TI - Bacteremia due to Campylobacter sputorum Biovar sputorum. PMID- 9868687 TI - Mucormycosis of petrous bone in an allogeneic stem cell transplant recipient. PMID- 9868688 TI - Primary cutaneous acanthamoeba infection in a patient with AIDS. PMID- 9868689 TI - Guillain-Barre polyneuropathy associated with mediterranean spotted fever: case report. PMID- 9868690 TI - Coagulase-negative staphylococcus endocarditis restricted to the normal pulmonic valve in a patient with end-stage renal disease: case report and review. PMID- 9868691 TI - Fatal encephalitis caused by Mycoplasma pneumoniae diagnosed by the polymerase chain reaction. PMID- 9868692 TI - Cardioverter-lead electrode infection due to Corynebacterium amycolatum. PMID- 9868693 TI - Treatment of endocarditis due to vancomycin-resistant Enterococcus faecium with quinupristin/dalfopristin, doxycycline, and rifampin: a synergistic drug combination. PMID- 9868694 TI - Primary meningococcal conjunctivitis in a human immunodeficiency virus-infected adult. PMID- 9868695 TI - Successful treatment of Kaposi's sarcoma with a combination of antiviral drug therapy and chemotherapy: two case reports. PMID- 9868696 TI - Increased serum procalcitonin levels are not specific to sepsis in neonates. PMID- 9868697 TI - Necrotizing fasciitis with Klebsiella liver abscess. PMID- 9868698 TI - Abatement of cutaneous Kaposi's sarcoma associated with cidofovir treatment. PMID- 9868699 TI - Prevaccination screening for hepatitis B among sexually active adolescents and young adults. PMID- 9868700 TI - Motility of the canine pyloric ring following a pylorus-preserving pancreatoduodenectomy. AB - Recently, function-preserving operations have become popular, and pylorus preserving pancreatoduodenectomy (PPPD) is frequently performed for diseases of the head of the pancreas. However, there are only a few basic studies on the pyloric function after PPPD. Using strain gauge force transducers (SGTs), we studied the pyloric motility of normal and PPPD dogs. We prepared three normal and three PPPD dogs in which the SGTs were implanted onto the antrum and pyloric ring, etc. In conscious dogs, the spontaneous gastrointestinal motility was recorded, and the plasma motilin concentration was measured during the interdigestive state. Following the administration of exogenous Leu13-motilin, the motility was again recorded. The relaxation and opening of the pyloric ring was observed synchronously with intense contractions of the antrum during the phase III of normal dogs. Phase III-like motility was recorded in the PPPD dogs, which was not a typical periodic motility. The plasma motilin concentration of one PPPD dog could be measured, and the motilin levels during the phase III-like motility were higher than during phase I. The phase III-like motility was induced by Leu13-motilin in both normal and PPPD dogs. The phase III-like motility recorded in the PPPD dogs was not a typical periodic one, and this aberrant motility was considered to be one of the causes of delayed gastric emptying. Phase III-like motility was induced by the administration of Leu13-motilin; therefore, it is possible that Leu13-motilin improved the motility of the pyloric ring after PPPD. PMID- 9868701 TI - Effects of excess K+ on carbachol-induced contractions in the guinea-pig tracheal muscle. AB - 1. In smooth muscles isolated from the guinea-pig trachea, the effects of dihydropyridines, nifedipine and nicardipine on contractions produced by carbachol (Cch) were studied in normal (6 mM) and excess K+ concentration (60 mM). The tonic contraction produced by 1 microM Cch was highly dependent on the external Ca2+ concentration ([Ca2+]0) and was not significantly affected by cyclopiazonic acid or thapsigargin, Ca2+ uptake inhibitor. 2. [Ca2+]0-tension curves were steeper in the presence of 1 microM Cch (the Hill coefficient: 2.5) than in the presence of 60 mM K+ (Hill coefficient: 1.6) and their ED50 of Ca2+ was 0.16 and 0.39 mM, respectively. An increase of K+ to 60 mM in the presence of 1 microM Cch shifted the curve to the left roughly in parallel (ED50: 0.12 mM, Hill coefficient: 2.3). 3. [Ca2+]0-tension curve in the presence of 1 microM Cch was shifted to the right in parallel by nifedipine (1 microM). This was markedly potentiated by 60 mM K+ (the increase in ED50 of Ca2+ being 3 times at 6 mM and 15 times at 60 mM K+). No tension was evoked by Ca2+ up to 2.5 mM in 60 mM K+ solution containing 1 microM nifedipine but no Cch. 4. In the absence of nifedipine, Cch-induced contractions were potentiated by 60 mM K+, whereas in the presence of nifedipine, Cch-induced contractions were markedly inhibited by 60 mM K+. These mechanical changes were accompanied by an increase or a decrease in intracellular Ca2+. 5. A hypothesis is presented to explain the results which suggests that the kinetics of Ca2+ influx though a single type of pathway is modulated by membrane potential and receptor activation and that the susceptibility of the pathway to dihydropyridine blockade is closely related to the Ca2+ influx kinetics with receptor activation reducing and membrane depolarization increasing the susceptibility. PMID- 9868702 TI - Simultaneous activation of Ca(2+)-dependent K+ and Cl- currents by various forms of stimulation in the membrane of smooth muscle cells from the rabbit basilar artery. AB - In smooth muscle cells isolated from cerebral blood vessels, histamine activates Cl- channels through an elevation of intracellular Ca2+. We investigated whether Cl- currents were also evoked by adenosine triphosphate (ATP) or caffeine in isolated smooth muscle cells from the rabbit basilar artery using the perforated patch-clamp technique. Bath application of 10 microM ATP or 1 mM caffeine (holding potential -60 mV) activated transient inward currents. With prolonged bath application of 10 microM ATP or 1 mM caffeine, oscillatory inward current were sporadically generated. At a holding potential of -40 mV, transient Cl- currents were induced by 10 microM histamine, 10 microM ATP, and 1 mM caffeine, following activation of a K+ current. At -10 or -20 mV, histamine predominantly activated the K+ current. A repetitively activated outward current was induced by membrane depolarization. These results suggest that oscillations in intracellular Ca2+ induced by histamine, ATP, and caffeine caused Cl(-)-current activation at the resting membrane potential. This Cl- current may depolarize the membrane and, thus activate voltage-dependent currents, including a Ca(2+)-dependent K+ current. Both the Ca(2+)-dependent K+ and Cl- currents induced by various stimuli may contribute to the modulation of Ca2+ influx by reinforcing membrane depolarization. PMID- 9868703 TI - Potentiating actions of lanthanum on ACh-induced cation current in guinea-pig ileal smooth muscle cells. AB - Potentiating actions of external lanthanum (La3+) on muscarinic receptor activated nonselective cation current (Icat) were investigated in myocytes dissociated from the longitudinal muscle layer of guinea-pig ileum, with a whole cell variant of the patch clamp technique. Icat was dissected from other membrane currents by loading Cs-aspartate into the cell. Application of submilimolar concentrations of La3+ following 300 microM ACh into the bath caused a dose dependent increase in the amplitude of Icat. The apparent Kd value for this increase was 190 microM, with a cooperativity factor of 1.7. La(3+)-induced increase in Icat amplitude was not associated with either changes in the reversal potential of Icat or altered sensitivity of muscarinic receptor to ACh, and paralleled by the conductance increase of Icat, the maximum of which (Gmax) occurred at about 1 mM La3+. Voltage-jump experiments revealed that the rate of current relaxation at hyperpolarizing potentials was greatly reduced in the presence of La3+, and correspondingly the steady state activation curve shifted toward more negative potentials. Divalent cations such as Cd2+ or Ni2+, which have been known to block Icat, antagonized the augmentative effect of La3+ on Icat in a competitive fashion, suggesting that the site of their actions might be similar. Furthermore, single Icat activities induced by internal perfusion of GTP gamma S (100 microM) was also greatly enhanced by external addition of 1 mM La3+. Under current clamp conditions, 1 mM La3+ blocked spontaneous Ca2+ spike activities, but was almost without effect on the membrane depolarization induced by ACh. In contrast, milimolar concentrations of Cd2+ and Ni2+ abolished both Ca2+ spike activities and ACh-induced depolarization. Potential importance of La3+ as a tool to investigate the external Ca(2+)-dependence of Icat has been discussed. PMID- 9868704 TI - Gastric emptying of three different size of indigestible radiopaque markers in healthy subjects. AB - The gastric emptying of indigestible solids is affected by the size, specific gravity, shape, consistency and the compressibility of the indigestible solids and depends largely on the composition of the meal. The threshold size is about 3.0 mm in canine. But it is reported that the estimates made for dogs may not apply to humans. I evaluated whether the size of indigestible solids affects gastric emptying in healthy subjects with food which had the same specific gravity as the indigestible solids. We used three sets of 20 ring shaped radiopaque markers, which had the same specific gravity (1.2), shape, consistency, and compressibility, but different diameters (2.0 mm, 4.5 mm and 7.0 mm). After the subjects ingested the three sets of 20 markers with food, the number of markers evacuated from the stomach was counted every 15 min until all the markers were evacuated. The median numbers of discharged markers of 2.0 mm, 4.5 mm and 7.0 mm were 5 (0-5), 1 (0-3) and 3 (0-4) at 60 min, 15 (3-18), 10 (6 9) and 13 (11-16) at 120 min (median, 25th-75th percentile), respectively. There was no statistically significant difference between the numbers of the three types of discharged markers during observation. The lag time (the period from the time of ingestion of the markers to the time when the first marker left the stomach) of the 2.0 mm, 4.5 mm and 7.0 mm markers was 52.5 (45-105), 67.5 (60 105), and 52.5 (45-105) min, respectively, and there was no statistically significant difference. In conclusion, the radiopaque markers of different diameters (below 7.0 mm) left the stomach in the same pattern. PMID- 9868705 TI - [Safety of radiotherapy]. PMID- 9868706 TI - [O-15-butanol Pet activation study on the cerebral representation of declarative memory]. AB - AIM: In this study, neuroanatomical correlates of encoding and retrieval in paired associate learning were evaluated with positron emission tomography using auditorily presented highly imaginable words. METHODS: Six right-handed normal male volunteers took part in the study. Each subject underwent six O-15-butanol PET scans. On each of the six trials the memory task began with the injection of a bolus of O-15-butanol. The subjects had to learn and retrieve twelve word pairs (highly imaginable words, not semantically related). The presentation of nonsense words served as reference condition. RESULTS: Recall accuracy after 2-4 presentations was high during the PET measurement. In both encoding and retrieval we found anterior cingulate activation. We show bilateral dorsolateral prefrontal activation during the encoding of auditorily presented word pair associates, whereas retrieval led to left frontal activation. Furthermore, we demonstrate the importance of the precuneus in the retrieval of highly imaginable word-pair associates. CONCLUSION: Our results support the hypothesis of the presence of distributed widespread brain structures subserving episodic declarative memory. PMID- 9868707 TI - [Investigations on the effect of "ecstasy" on cerebral glucose metabolism: an 18 FDG PET study]. AB - PURPOSE: The aim of the present study was to determine the acute effects of the "Ecstasy" analogue MDE (3, 4-methylendioxyethamphetamine) on the cerebral glucose metabolism (rMRGlu) of healthy volunteers. METHOD: In a randomised double-blind trial, 16 healthy volunteers without a history of drug abuse were examined with 18-FDG PET 110-120 minutes after oral administration of 2 mg/kg MDE (n = 8) or placebo (n = 8). Beginning two minutes prior to radiotracer injection, a constant cognitive stimulation was maintained for 32 minutes using a word repetition paradigm in order to ensure constant and comparable mental conditions during cerebral 18-FDG uptake. Individual brain anatomy was represented using T1 weighted 3D flash MRI, followed by manual regionalisation into 108 regions-of interest and PFT/MRI overlay. Absolute quantification of rMRGlu and comparison of glucose metabolism under MDE versus placebo were performed using Mann-Whitney U test. RESULTS: Absolute global MRGlu was not significantly changed under MDE versus placebo (MDE: 41.8 +/- 11.1 mumol/min/100 g, placebo: 50.1 +/- 18.1 mumol/min/100 g, p = 0.298). The normalised regional metabolic data showed a significantly decreased rMRGlu in the bilateral frontal cortex: left frontal posterior (-7.1%, p < 0.05) and right prefrontal superior (-4.6%, p < 0.05). On the other hand, rMRGlu was significantly increased in the bilateral cerebellum (right: +10.1%, p < 0.05; left +7.6%, p < 0.05) and in the right putamen (+6.2%, p < 0.05). CONCLUSIONS: The present study revealed acute neurometabolic changes under the "Ecstasy" analogon MDE indicating a fronto-striato-cerebellar dysbalance with parallels to other psychotropic substances and various endogenous psychoses respectively. PMID- 9868708 TI - Paradoxical hypotension during dobutamine infusion for myocardial perfusion scintigraphy. AB - AIM: Dobutamine as a predominant beta-1 agonist increases heart rate and myocardial contractility and at sufficient high doses, it also increases systolic blood pressure. This study was undertaken to describe instances of paradoxical hypotension during dobutamine infusion for TI-201 myocardial perfusion SPECT study and the relationship between scintigraphic findings and hypotension occurred during dobutamine infusion. METHODS: In 201 consecutive patients unable to perform adequate exercise, dobutamine TI-201 myocardial SPECT was performed. Dobutamine was infused starting from 10 micrograms/kg/min increasing to 40 micrograms/kg/min. Paradoxical hypotension was defined as a decrease in systolic blood pressure > or = 20 mmHg compared with baseline study. RESULTS: Paradoxical hypotension was observed in 40 patients (Group A) out of 201 (19.9%) while no significant change in systolic blood pressure was detected in the remaining 161 patients (Group B). Mean maximum fall in systolic blood pressure was 39 +/- 18 mmHg (range: 20-90). In 33 of 40 patients (83%) with paradoxical hypotension, scintigraphy was normal compared to 131 (81%) of the remaining 161 patients. In patients of Group A, angiography, echocardiography and tilt table tests were performed in 13, 11 and 6 patients respectively. Nine of 13 angiographic evaluations (69%), 10 of 11 echocardiographic evaluations (91%), all of the tilt table tests were normal. Additionally, all of the patients of Group A were clinically followed up at least 6 months after the myocardial perfusion scintigraphy. None of the patients had a cardiac event except one patient during the follow-up period. CONCLUSION: Paradoxical hypotension during dobutamine infusion for myocardial scintigraphy is not an uncommon finding and up to 19.9% patients may develop such hypotension. To maximize test safety, precautions should be taken during dobutamine myocardial stress test, since remarkable decrease in systolic blood pressure may occur. Unlike hypotension occurring with exercise test, hypotension response to dobutamine is not always a marker for coronary artery disease. PMID- 9868709 TI - Arthroscintigraphy in suspected rotator cuff rupture. AB - AIM: In order to evaluate the diagnostic efficiency of arthroscintigraphy in suspected rotator cuff ruptures this new imaging procedure was performed 20 times in 17 patients with clinical signs of a rotator cuff lesion. The scintigraphic results were compared with sonography (n = 20), contrast arthrography (n = 20) and arthroscopy (n = 10) of the shoulder joint. METHODS: After performing a standard bone scintigraphy with intravenous application of 300 MBq 99m-Tc methylene diphosphonate (MDP) for landmarking of the shoulder region arthroscintigraphy was performed after an intraarticular injection of 99m-Tc microcolloid (ALBU-RES 400 MicroCi/5 ml). The application was performed either in direct combination with contrast arthrography (n = 10) or ultrasound conducted mixed with a local anesthetic (n = 10). Findings at arthroscopical surgery (n = 10) were used as the gold standard. RESULTS: In case of complete rotator cuff rupture (n = 5), arthroscintigraphy and radiographic arthrography were identical in 5/5. In one patient with advanced degenerative alterations of the shoulder joint radiographic arthrography incorrectly showed a complete rupture which was not seen by arthroscintigraphy and endoscopy. In 3 patients with incomplete rupture, 2/3 results were consistent. A difference was seen in one patient with a rotator cuff, that has been already revised in the past and that suffered of capsulitis and calcification. CONCLUSION: Arthroscintigraphy is a sensitive technique for detection of rotator cuff ruptures. Because of the lower viscosity of the active compound, small ruptures can be easily detected, offering additional value over radiographic arthrography and ultrasound, especially for evaluation of incomplete cuff ruptures. PMID- 9868710 TI - Limited value of fluorine-18-fluorodeoxyglucose PET for the differential diagnosis of focal liver lesions in patients with chronic hepatitis C virus infection. AB - AIM: The differentiation of HCC from liver metastasis or benign disorders by imaging studies based upon morphological aspects may be difficult. METHOD: In order to evaluate the role of tumour metabolism, we performed FDG-PET (whole-body PET and transmission-corrected regional scans of the liver as well as the SUV determined 60 min after injection of FDG) in ten consecutive patients with HCV associated focal liver lesions. Definite diagnosis was established after ultrasound-guided liver biopsy followed by histopathological examination. These results were compared with ultrasound, computed tomography, serum anti-p53, and p53 protein expression. RESULTS: The histologic examination revealed a HCC in five patients, regenerative nodules in three patients, and liver metastasis (primary malignancy: one adenocarcinoma and one neuroendocrine tumour) in the remaining two patients. Three of ten lesions were detectable by FDG-PET: two HCCs and one metastatic adenocarcinoma. Seven lesions were not distinguishable by FDG PET (three HCCs, three regeneration nodules and one metastatic neuroendocrine tumour). In each patient hepatic lesions were visible either by ultrasound or CT. Both tumours (metastatic adenocarcinoma, moderately well-differentiated HCC) with the strongest expression of p53 also presented with highly increased FDG uptake. CONCLUSIONS: FDG-PET is not superior to ultrasound or CT and therefore does not allow the non-invasive differentiation of HCV-associated focal liver lesions. Tissue-diagnosis by means of liver-biopsy followed by histopathological examination remains the gold-standard for the differentiation of HCV-related liver lesions. The finding of the relationship of p53 protein overexpression with the SUV needs further confirmation. PMID- 9868711 TI - [Influence of disease prognosis and age on the radiation risk from diagnostic nuclear medicine (bone scintigraphy as an example)]. AB - AIM: Of the study was to assess the influence of the patient age and the survival of patients undergoing bone scintigraphy on the calculations of the theoretical lifetime loss. PATIENTS AND METHODS: The evaluated data set included 216 patients undergoing a bone scan for the first time in 1980. From 182 patients a study end point (either the date of death, or existing registration in the German resident office on the 31.12.1995) was obtained. The theoretical life time loss was based on the formalism previously presented by Schicha und Wellner (13). RESULTS: In 74% of the examined patients a malignant disease was present. 28% of the patients had died within one year of the examination. At the end of the follow-up period, of at least 15 years, 58% of the patients were deceased. The theoretical lifetime loss due to radiation exposure within this specified patient group was estimated to be 0.152 d/mSv. Assuming an age distribution of the general population and statistically derived life expectancy, this value should be 0.437 d/mSv. CONCLUSION: The estimates regarding the risk of diagnostic procedures using ionizing radiation should take into account the limited life expectancy of patients in hospitals. PMID- 9868712 TI - Cold metastases detected by bone scintigraphy in aggressive lung cancer. AB - A case of a 55-year-old man was remitted to Traumatology Department to present back pain of two weeks of evolution. The results of bone scintigraphy and the patient's evolution allowed the diagnosis. This case report and a literature review showed the importance of using a routine bone scan in diagnosis of bone metastases. PMID- 9868713 TI - Human chorionic gonadotropin and CA 15-3 producing adenocarcinoma. AB - 50 years old man suffering from primary lung adenocarcinoma presented with high levels of both beta subunit human chorionic gonadotropin (beta HCG) and cancer antigen 15-3 (CA 15-3) in the absence of elevated carcinoembrionic antigen (CEA), alfa fetoprotein (AFP) and carbohydrate antigen 19-9 (CA 19-9). Although beta HCG or CA 15-3 high levels were reported in adenocarcinoma of lung, this is the first report of a patient with high levels of both markers. PMID- 9868714 TI - Formalin mishandling during wisdom tooth surgery. PMID- 9868715 TI - Infection control practices of oral specialists and general dental practitioners. PMID- 9868716 TI - Concurrent cervical and craniofacial pain. A review of empiric and basic science evidence. AB - Because many patients present themselves for treatment with both craniofacial and craniocervical pain, 2 questions arise: (1) What are the sensory and motor consequences of dysfunction in either of these areas on the other? (2) Do craniofacial and craniocervical pain have a similar cause? These questions formed the impetus for this review article. The phenomenon of concurrent pain in craniofacial and cervical structures is considered, and clinical reports and opinions are presented regarding theories of cervical-to-craniofacial and craniofacial-to-cervical pain referral. Because pain referral between these 2 areas requires anatomic and functional connectivity between trigeminally and cervically innervated structures, basic neurophysiologic and neuroanatomic literature is reviewed. The published data clearly demonstrate neurophysiologic and structural convergence of cervical sensory and muscle afferent inputs onto trigeminal subnucleus caudalis nociceptive and non-nociceptive neurons. Moreover, changes in metabolic activity and blood flow in the brainstem and cervical dorsal horn of the spinal cord in both monkeys and cats have been demonstrated after electric stimulation of the V1-innervated superior sagittal sinus. In conclusion, the animal experimental data support the findings of human empiric and experimental studies, which suggest that strong connectivity exists between trigeminal and cervical motor and sensory responses. PMID- 9868717 TI - Lingual ulceration. PMID- 9868718 TI - Edema in the upper airway in patients with obstructive sleep apnea syndrome. AB - OBJECTIVE: The purpose of this study was to investigate the histologic alterations in the upper airway in patients with obstructive sleep apnea syndrome. STUDY DESIGN: Surgical specimens were obtained during non-laser assisted uvulopalatopharyngoplasty from 5 adult patients (4 male and 1 female) with polysomnographically confirmed obstructive sleep apnea syndrome. RESULTS: In all of the specimens, interstitial edema was present with mild inflammatory reaction. Fat was not at all prominent. CONCLUSIONS: It is confirmed histologically that there is edema present in the upper airway in patients with obstructive sleep apnea syndrome. It is uncertain whether this edema is one of the causative mechanisms of the obstruction or a consequence of the syndrome. PMID- 9868719 TI - An in vitro evaluation of distractors used for osteogenesis. AB - OBJECTIVE: The objective of this investigation was to evaluate and compare the mechanical behaviors of distractors used for osteogenesis under various conditions by means of common engineering standards. STUDY DESIGN: Five groups of 5 synthetic mandibles were used in this study (N = 25). The first portion of the investigation compared mandibles without intervention (group A [controls]; n = 5), mandibles that had uniform osteotomies stabilized with an external distractor (group B; n = 5), and mandibles that had simulated sagittal osteotomies rigidly fixated with 3 positional screws (group C; n = 5). The second portion of the investigation compared uniform osteotomies (group B; n = 5) and uniform corticotomies (group D; n = 5) that were stabilized with the same external distractor. The last portion of the investigation compared osteotomies stabilized with an external distractor (group B; n = 5) and osteotomies stabilized with an internal distractor (group E; n = 5). Each construct was subjected to vertical loads on a mechanical testing unit. Common engineering standards, including yield load, yield displacement, maximum load, displacement at maximum load, and stiffness, were measured, recorded, and compared by means of a 1-way analysis of variance and a Scheffe multiple comparison test or independent-samples t test. The means between groups were considered significant for P < .05. A polynomial best-fit curve was calculated for the load/displacement data for each group. RESULTS: During the first portion of the investigation, no significant differences were noted between the control, rigidly fixated sagittal osteotomy, and external distractor with osteotomy groups for displacement at maximum load (P = .19). Significant differences were noted between groups for yield displacement (P = .009), yield load (P < .001), maximum load (P < .001), and stiffness (P < .007). Failures occurred in the control and rigidly fixated groups with fractures of the synthetic mandibles. Failures occurred in the external distractor group with permanent deformation or torsion of the pins. During the second portion of the experiment, no statistically significant differences were noted between the corticotomy and osteotomy groups in stiffness (P = .363), maximum load (P = .207), or yield displacement (P = .940). Statistically significant differences were noted between groups for yield load (P = .036) and displacement at maximum load (P = .010). Failures occurred in both groups with permanent deformation or torsion of the pins. During the last portion of the investigation, statistically significant differences were noted between the external distractor and internal distractor groups in yield load (P < .001), yield displacement (P < .001), maximum load (P = .001), and displacement at maximum load (P = .01); no significant differences were noted in stiffness (P = .71). Failures occurred in the external distractor group with permanent deformation or torsion of the pins. Failures occurred in the internal distractor group with fracture of the model or displacement beyond 30.0 mm. CONCLUSIONS: Different patterns of mechanical behavior were found between the control and rigidly fixated sagittal osteotomy groups and the external distractor group, between the corticotomy and osteotomy groups, and between the internal and external distractor groups. PMID- 9868720 TI - Cryosurgical treatment of melanin-pigmented gingiva. AB - OBJECTIVE: Abnormal deposition of melanin in gingiva can often occur. Complaints of "black gums" are common, and demand for depigmentation is usually made for esthetic reasons. A simple and effective cryosurgical technique to eliminate the pigmentation is the subject of this article. STUDY DESIGN: Twenty patients with dark gingiva were treated by direct application of liquid nitrogen with a cotton swab for 20 to 30 seconds. RESULTS: The treated gingiva appeared normal within 1 to 2 weeks after 1 or 2 cryosurgical treatments. The acceptance of the treatment was excellent. CONCLUSIONS: This was a simple, bloodless cryosurgery for the depigmentation of gingiva, requiring no local anesthesia or sophisticated equipment. PMID- 9868721 TI - Synovial chondromatosis of the temporomandibular joint with intracranial extension. AB - An unusual case of synovial chondromatosis of the temporomandibular joint with intracranial extension, which resulted in complete dehiscence of the floor of the middle cranial fossa, is reported. An overview of the current literature and a discussion of the diagnosis and surgical management of synovial chondromatosis are presented. PMID- 9868722 TI - Primary temporary AO plate reconstruction of the mandible. AB - OBJECTIVE: This study assessed the incidence of complications and revisions after primary temporary AO plate reconstructions of the mandible performed from 1971 through 1996. STUDY DESIGN: In a retrospective record review, the data of 51 patients undergoing primary temporary AO plate reconstructions after composite mandibular resection were analyzed according to age, gender, date of reconstruction, anatomic location of reconstruction, use of additional radiotherapy and/or flap surgery, and incidence of associated complications and revisions. RESULTS: Associated complications and revisions accompanying primary temporary AO plate reconstructions were evaluated through use of the incidence rates of associated complications (IACs) and revisions (IARs), which are reported as the numbers of associated complications and revisions, respectively, per 100 patients. Of the entire sample, the highest incidence rate of associated complications was associated with infection (33.5); this was followed by the rates for plate exposure (27.9) and plate fracture (10.7). With an overall incidence rate of 38.3 revisions per 100 patients, the corresponding site-related incidence rates of associated revisions were found to be 55.0 for reconstructions of the anterior mandible crossing the midline, 37.1 for those of the body segment of the mandible, and 31.1 for reconstructions involving the ramus and/or condyle area of the mandible. Revision rates were observed to be significantly higher in irradiated (53.6 vs 31.5) and non-flap-added reconstructions (43.2 vs 24.8). Analysis of treatment group-related incidences revealed irradiated non-flap-added reconstructions to be associated with the highest failure rates, whereas additional flap surgery resulted in a significant reduction in complications (50.0 vs 108.0) and revisions (20.0 vs 65.7). CONCLUSIONS: This study showed primary temporary AO plate reconstructions after composite mandibular resection to be associated with a high rate of complications and revisions. The results emphasize the need to relate outcome measures to site-related and treatment related parameters. PMID- 9868723 TI - Periodontal attachment loss in patients after head and neck radiation therapy. AB - OBJECTIVE: The purpose of this study was to determine the potential impact of head and neck radiation therapy on the progression of periodontal attachment loss. STUDY DESIGN: Ten patients who received unilateral radiation fields that included the dentition were assessed before radiation treatment and after irradiation at a mean age of 6.01 years. Complete oral, dental, and periodontal examinations were completed by one examiner. The results were assessed through use of paired t tests. RESULTS: More teeth were extracted because of periodontal disease in the field of radiation after irradiation. Remaining teeth in the radiated volume showed an increase in probing depth of 0.82 mm in comparison with 0.40 mm for teeth in the nonradiated region (P = .05). Recession on the facial aspects was 1.88 mm for teeth in the radiated volume and 1.16 mm for teeth in the nonradiated region (P = .001), and recession on the lingual aspects was 2.10 for teeth in the radiated volume and 0.91 for teeth in the nonradiated region (P = .05). Mean total attachment loss was 2.81 mm for teeth in the radiated sites; this compared with 1.43 mm for teeth in the nonradiated sites (P = .003). Increased mobility of teeth in the high-dose fields was seen (P = .02). CONCLUSIONS: This study showed that tooth loss and greater periodontal attachment loss occur in teeth that are included within high-dose radiated sites of patients treated with irradiation therapy for cancer. These findings should be considered in preradiation treatment planning. PMID- 9868724 TI - Methodologic considerations for research in traditional (alternative) medicine. AB - The widepsread use, in the United States and other western countries, of alternative medical practices often derived from traditional systems of health has intensified the need to evaluate their safety and effectiveness. These practices tend to generate polarized attitudes of either acceptance or rejections. Opponents reject them because, they claim, good scientific research has not supported their use. Proponents insist that current scientific methodologies are not appropriate to evaluate these practices. This article briefly examines these issue, suggesting that the answer lies between these 2 extreme positions. Various methodologic issues are considered, and suggestions are made to adapt scientific methodology, where appropriate, to respect the particularities of alternative medicine. PMID- 9868726 TI - Psychologic factors and oral lichen planus. A psychometric evaluation of 100 cases. AB - OBJECTIVE: The purpose of this study was to analyze the importance of psychologic factors in patients with oral lichen planus, and attempts were made to identify possible personality features characteristic of patients with oral lichen planus. STUDY DESIGN: The study involved 100 patients with oral lichen planus (group 1) and 50 control subjects (group 2). We applied the following psychometric tests to both groups: Spielberger State-Trait Anxiety Inventory, Cattell Personality Questionnaire 16PF, Hassanyeh Rating of Anxiety-Depression-Vulnerability, Beck Depression Inventory, Raskin Depression Screen, and Covi Anxiety Screen. RESULTS: The patients with oral lichen planus were found to exhibit greater anxiety, as reflected by statistically significant scores with the anxiety tests that were used (Spielberger State-Trait Anxiety Inventory, Cattell Personality Questionnaire 16PF, Hassanyeh Rating of Anxiety-Depression-Vulnerability, and Covi Anxiety Screen). The patients with oral lichen planus likewise exhibited greater depression than the controls in all 3 depression tests applied (Beck Depression Inventory, Hassanyeh Rating of Anxiety-Depression-Vulnerability, and Raskin Depression Screen) and were more vulnerable to psychic disorders on the basis of the PD subscales (vulnerability) of the Hassanyeh questionnaire. Three features (conformity to the group, astuteness, and rebelliousness) defined the personalities of our patients with oral lichen planus, according to the Cattell 16PF questionnaire. Finally, those patients with erosive lichen planus exhibited higher depression scores than patients with nonerosive lichen planus. CONCLUSIONS: Despite the higher anxiety scores observed in patients with oral lichen planus, it was not established that the observed psychologic alterations constitute a direct etiologic factor of oral lichen planus; nor was it established that such alterations are a consequence of oral lichen planus and its lesions. PMID- 9868725 TI - Effectiveness of kampo medicine (sai-boku-to) in treatment of patients with glossodynia. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of a Kampo medicine (Sai-boku-to) for treatment of patients with glossodynia. STUDY DESIGN: Sai-boku-to or an antianxiety drug (diazepam) with vitamin B complex was administered orally for 3 months to each of 200 patients with glossodynia. Clinical examination evaluated the following subjective symptoms: pain, burning sensation, and discomfort. Effectiveness was evaluated as follows: "markedly effective," all 3 symptoms disappeared; "effective," pain improved; "ineffective," no improvement in pain. RESULTS: The effective rates were 70% after 1 month, 85% after 2 months, and 92% after 3 months of administration of Sai-boku-to (the Kampo group) and 74% after 1 month, 71% after 2 months, and 69% after 3 months of administration of the antianxiety drug with vitamin B complex (the control group). No significant side effect was noted in the Kampo group, but sleepiness was recorded in 33 cases in the control group. CONCLUSIONS: The results indicate that Sai-boku-to may be a clinically useful medicine for the long-term treatment of patients with glossodynia. PMID- 9868727 TI - Graft-versus-host disease. Case report and discussion. AB - The graft-versus-host disease reaction is an immunologic consequence resulting from the grafting of immunocompetent cells from one individual to an immunocompromised host. The oral manifestations of this disease process include severe oral pain, xerostemia, ulcerative lesions, and mucositis. With appropriate history and diagnostic testing, these lesions can be differentiated from other oral diseases having similar clinical appearance. PMID- 9868728 TI - Oral cancer in a tobacco-chewing US population--no apparent increased incidence or mortality. AB - OBJECTIVE: Smokeless tobacco use is an accepted risk factor for oral cancer in the United States, but the major proof of this is based largely on a single epidemiologic (case-control) comparison of women, whereas the chewing of tobacco is predominantly a habit of men. The present investigation sought to compare gender-specific oral cancer mortality and incidence rates in West Virginia, the state with the highest per capita consumption of smokeless tobacco, with rates from other states and with the US average rates. It was hypothesized that the cancer rates for West Virginia males would be significantly greater than the US average and greater than the rates for states with less smokeless tobacco consumption. STUDY DESIGN: Data from the West Virginia Cancer Registry for the years 1993 through 1995 were compared with data from contemporary Surveillance and Epidemiology End Results for the US (with respect to incidence) and from the Center for Disease Control and Prevention's National Center for Health Statistics (with respect to mortality). RESULTS: The average annual incidence rates (per 100,000 population) for oral/pharyngeal cancer in West Virginia males and females were 13.4 and 5.1, respectively; these compared with rates of 15.4 and 5.7 for the US. The average annual mortality rates (per 100,000 population) for the disease in West Virginia males and females were 4.2 and 1.6, respectively; these compared with 4.4 and 1.5 for the US. West Virginia oral/pharyngeal mortality rates for both genders were statistically significantly lower than US rates throughout the years 1950 through 1980. Among other potential oral cancer etiologic factors, the very low prevalence of alcohol abuse in West Virginia seemed to be relevant. CONCLUSIONS: The hypothesis was not confirmed by data analysis. West Virginia is the state with the highest per capita consumption of smokeless tobacco, yet it has less oral/pharyngeal cancer than the US average. The authors strongly urge additional and improved epidemiologic evaluation of the oral cancer risk of smokeless tobacco use in US males. PMID- 9868729 TI - Odontogenic tumors. A demographic study of 759 cases in a Chinese population. AB - Seven hundred fifty-nine cases of odontogenic tumors retrieved from the files of College of Stomatology, West China University of Medical Sciences were classified according to the World Health Organization's Histological Classification of Odontogenic Tumors and compared with similar reports from other countries. Among these cases, 93.9% of the tumors were benign and 6.1% were malignant. Ameloblastomas predominated (58.6%) with a predilection for the mandible, while odontomas, generally regarded as the most frequent odontogenic tumor in North America, only accounted for 6.7%, the fourth most common tumor in this series. The mandible and the maxilla were divided into eight anatomic regions, and the distribution of each odontogenic tumor type amongst these regions was recorded. The relative incidence of each tumor type, patient age and gender were also compared with data from other selected large series. Geographic differences were noted in the relative incidence of ameloblastoma, odontoma and malignant odontogenic tumors among the Chinese/African, North American and Turkish/German groups. Ameloblastoma and malignant odontogenic tumors are not considered rare in a Chinese population. PMID- 9868730 TI - Melanoma arising from the mucosal surfaces of the head and neck. AB - Oral mucosal malignant melanoma is a rare disease. We reviewed 30 years of data from a tumor registry and identified 65 patients who had head and neck melanomas. Two thirds (43) of the 65 patients were identified as male, with the mean age in the sixth decade. Of the 65 patients, only 6 had melanoma that arose from the oropharyngeal mucosa. Of the lesions involving the oral mucosa, each lesion manifested itself as a mass or was associated with symptoms of discomfort; only one third (2) of the lesions were pigmented. The clinician must carefully examine the head, neck, and oral cavity, and any pigmented lesion that is not recognized as a specific entity, such as amalgam tattoo, should be biopsied. The more common presentation of amelanotic malignant melanoma requires a high index of suspicion for masses identified in the mouth and requires biopsy for definitive diagnosis. The prognosis for oral mucosal malignant melanoma is poor. PMID- 9868731 TI - Oncocytoid artifact of the parotid gland: a newly reported artifact. AB - Electrocautery can induce significant alterations in the connective tissues and epithelium of specimens removed for diagnostic or therapeutic purposes. When electrocautery is used during parotid surgery, it can cause an oncocytoid artifact. The alterations described in this article are enlarged, tightly packed serous acinar cells with coarse to granular eosinophilic cytoplasm, distinct cell borders, and round basal nuclei that on cursory microscopic examination resemble oncocytes with respect to morphology. These changes are seen in conjunction with other, more recognized changes secondary to electrocautery and are believed to occur as a consequence of the electrothermal discharge. On the basis of our findings, this artifact is common is parotid surgical specimens and was misdiagnosed as benign oncocytic lesions in 5 cases. PMID- 9868732 TI - Use of a hydroxylapatite-based material and calcium sulfate as artificial floors to repair furcal perforations. AB - OBJECTIVE: The purpose of this study was to evaluate a hydroxylapatite-based material and calcium sulfate when each was used under a resin-modified glass ionomer cement to repair furcation perforations. STUDY DESIGN: Perforations of pulp chamber floors were made in 72 teeth of 9 dogs. Perforations were divided into 3 equal-sized groups and repaired with resin-modified glass ionomer either alone or over an artificial floor. The artificial floor was either a hydroxylapatite-based material or calcium sulfate. Three dogs were killed at each of 3 intervals (1, 3, and 6 months). The tissue response to the tested materials was evaluated clinically, radiographically, and histologically. RESULTS: The hydroxylapatite-based material showed the highest radiographic success; this was followed by calcium sulfate and glass ionomer. From histologic evaluation, the average success rate was found to be 67% for calcium sulfate, 62% for the hydroxylapatite-based material, and 59% for glass ionomer. However, there was no statistical significant difference with the resin-modified glass ionomer when it was used alone and when it was used over a barrier. There was also no significant difference between the hydroxylapatite-based material and the calcium sulfate when they were used as artificial floors. CONCLUSION: The use of an artificial floor may not be necessary when flowable resin-modified glass ionomer cements are used. PMID- 9868733 TI - Prevalence of pulp stones in Jordanian adults. AB - OBJECTIVE: The purpose of this study was to describe the prevalence of pulp stones in a sample of Jordanian dental patients. The article also presents frequency distributions of this condition among different types of teeth. STUDY DESIGN: Data were collected through radiographic examinations of a random sample of periapical and bitewing films of 4573 teeth from 814 dental records from the Faculty of Dentistry's filing system. RESULTS: Pulp stones were identified in 22% of the teeth examined. First and second molars were the teeth most commonly affected. Pulp stone incidence was not significantly different among different age and gender groups. PMID- 9868734 TI - Dentinogenesis imperfecta: endodontic implications. Case report. AB - Dentinogenesis imperfecta is a hereditary disorder resulting in defective dentin in both the primary and secondary dentitions. The complications of dentinogenesis imperfecta are difficult to manage and provide a challenge to the dentist. This case report concerns treating an African American patient with dentinogenesis imperfecta who appeared for treatment with endodontic pathosis. It illustrates the need for appropriate and timely restorative treatment to prevent pulpal pathosis. Also demonstrated is the difficulty of endodontically treating dentinogenesis imperfecta teeth because of pulpal obliteration and abnormal dentin mineralization. Early and correct diagnosis of dentinogenesis imperfecta is imperative to enable appropriate preventive interventions and optimal dental treatment. Although pulpal pathosis is rarely reported with dentinogenesis imperfecta, endodontic treatment is occasionally necessary and has a guarded prognosis if initiated after pulp canal obliteration has occurred. PMID- 9868735 TI - Radiologic changes of the temporomandibular joint after condylar fractures in childhood. AB - OBJECTIVE: The purpose of this investigation was to study the radiologic outcomes of pediatric condylar fractures. STUDY DESIGN: Radiographs of 37 patients with 45 condylar fractures that had been diagnosed at the Department of Oral and Maxillofacial Surgery, Helsinki University Central Hospital were retrospectively reviewed. The frequency and characteristics of incomplete remodeling at an average of 4.1 years after the injury were studied. The influence of age and fracture type on remodeling were considered. RESULTS: Incomplete remodeling was observed after 56% of the fractures. The most frequent signs of incomplete remodeling were alteration in the configuration of the surface of the condylar head and deformation of the condylar neck. Incomplete remodeling was frequently (83%) related to fracture dislocation. For this fracture type the radiologic prognosis seems poor in all age groups. A difference in ramus height between the 2 sides was observed in 52% of the fractures, particularly frequently (80%) after fracture dislocation. CONCLUSIONS: Radiologic aberrations are frequent after pediatric condylar fractures. Dislocated fractures, in particular, need special attention and long-lasting clinical follow-up. PMID- 9868736 TI - Interobserver reliability in quantitative MRI assessment of temporomandibular joint disk status. AB - OBJECTIVE: The purpose of this study was to investigate interobserver reliability of a new technique for quantification of magnetic resonance images of temporomandibular joint disk status. STUDY DESIGN: Sixty magnetic resonance images of adolescent temporomandibular joints were randomly drawn for analysis. Four experienced observers traced the articular disk and osseous structures on sagittal magnetic resonance slice images. Quantitative measurements of disk length and disk displacement were recorded for each slice of 57 joints traced by each observer through use of a new quantification technique. Intraclass correlation coefficients were computed to assess interobserver agreement in the tracing of joint structures. RESULTS: The calculated intraclass correlation coefficient was 0.681 for disk length and 0.830 for disk displacement. In addition, the mean variability among observers was 1.041 mm for measurement of disk length and 0.972 mm for measurement of disk displacement. CONCLUSIONS: Interobserver agreement is high when the new quantification technique is used to interpret magnetic resonance images. PMID- 9868737 TI - Gas in the temporomandibular joint: computed tomography findings. Report of 3 cases. AB - In each of 3 patients referred for computed tomographic examinations of the temporomandibular joint, a gas collection was observed in the inferior space of the joint. The gas was observed on the posterosuperior surface of the mandibular condyle in the open mouth position in each patient. Each of 2 patients had saucer shaped bone resorption of the mandibular condyle adjacent to the gas; 1 patient had no remarkable bone abnormality. A history of temporomandibular joint puncture or facial trauma was denied by all 3 patients. PMID- 9868738 TI - Imaging findings of some buccomasseteric masses. AB - OBJECTIVE: The purpose of this study was to review and illustrate the appearances on diagnostic images of pathologic conditions of the buccomasseteric region. STUDY DESIGN: Computed tomographic, sonographic, and magnetic resonance images of the buccomasseteric masses were reviewed retrospectively. RESULTS: In our series of 66 cases, there were 60 benign lesions; of these, 21 were benign masses, 11 were inflammations, and 28 were myopathies. The most common pathologic condition of the buccomasseteric region was masseteric hypertrophy (n = 22). The most common lesion manifesting as a benign mass in this series was hemangioma (n = 10), which was followed by lipoma (n = 6). Six of the 66 patients had malignant tumors. CONCLUSIONS: Computed tomography and magnetic resonance imaging were useful in demonstrating the location and extent of the buccomasseteric masses. Sonography provided clear images of internal architectures of the mass in this region. PMID- 9868739 TI - Findings of clinical and radiographic caries among several adult age groups. AB - OBJECTIVE: The purpose of this study was to determine whether there are aged based variations in the association between clinically detected and radiographically detected caries and whether the prevalence of clinically undetected radiographic caries varies across adult age groups. STUDY DESIGN: The data for the analysis were from a clinical study that evaluated the efficacy of guidelines for prescribing dental radiographs. A total of 460 subjects had clinical examinations and interpretation made on full-mouth radiographs. Analysis was conducted to determine the tooth-specific and subject-specific prevalences of clinically undetected caries and to establish whether the association between clinical signs and radiographic signs varied by subject age. RESULTS: In total, approximately 5.8% of clinically sound teeth showed radiographic evidence of dentinal caries, and the prevalence increased with patient age. The prevalence of clinical signs of medium and large caries was 7.8% in 12,358 teeth in which caries was both clinically and radiographically present. However, for more than 80% of the teeth with clinically undetected caries, the lesions were evident on the interproximal radiographs. The associations between clinical and radiographic signs of dentinal caries were homogeneous across age groups. CONCLUSION: The findings demonstrate that adolescents and adults continue to have medium and large caries, although the location of the caries differs by age group, with higher proportions of gingival caries in older patients. PMID- 9868740 TI - Mechanism of early carbon tetrachloride toxicity in cultured rat hepatocytes. AB - CCl4 causes liver necrosis in a dose-dependent manner in vivo. However, we found that primary rat hepatocytes in culture were killed after a 2 hr incubation with carbon tetrachloride gas at CCl4 partial pressures above a threshold between 45 and 54 mmHg. Below this threshold concentration no increase in hepatocyte death was observed. We sought to explain the very abrupt CCl4 concentration threshold for hepatocyte death. Two inhibitors of cytochrome P450 2E1, cimetidine and diallyl sulfide, inhibited lipid peroxidation as measured by production of isoprostanes, but did not reduce hepatocyte death from CCl4. At 37 degrees, CCl4 accelerated the mitochondrial permeability transition in vitro, at a threshold CCl4 concentration similar to that which caused hepatocyte death. Phospholipase A2 inhibitors, mepacrine and 4-bromophenacyl bromide, inhibited the increase in mitochondrial permeability, but did not inhibit hepatocyte death caused by CCl4. Rat liver microsomal lipids were used to make liposomes loaded with Ponceau Red (FW 760.6). No leakage of Ponceau red was found at CCl4 concentrations greater than the threshold for cell death. However, CCl4 caused acceleration of liposome fusion, over the CCl4 concentration range spanning the threshold for hepatocyte death. Early hepatocyte death in cell culture is independent of metabolism of CCl4, and may be related to direct effects of CCl4 on intracellular membranes. PMID- 9868741 TI - Fluvoxamine is a potent inhibitor of the metabolism of caffeine in vitro. AB - The selective serotonin re-uptake inhibitor, fluvoxamine, is a very potent inhibitor of CYP1A2, and accordingly causes pharmacokinetic interactions with drugs metabolised by CYP1A2, such as caffeine, theophylline, imipramine, tacrine and clozapine. Interaction between caffeine and fluvoxamine has been described in vivo, leading to lowering of total clearance of caffeine by 80% during fluvoxamine intake. The main purpose of the present study was to evaluate this interaction in vitro in human liver microsomes. A high-performance liquid chromatography method was developed in order to assay 1,3-dimethylxanthine, 1,7 dimethylxanthine, 3,7-dimethylxanthine and 1,3,7-trimethyluric acid formed from caffeine by human liver microsomes. The limit of detection was 0.06 nmol.mg protein-1.hr-1. As expected, fluvoxamine was a very potent inhibitor of the formation of the N-demethylated caffeine metabolites, displaying Ki values of 0.08-0.28 microM. The formation of 1,7-dimethylxanthine was virtually abolished by 10 microM of fluvoxamine, indicating that the N3-demethylation of caffeine is almost exclusively catalysed by CYP1A2. The CYP3A4 inhibitors, ketoconazole and bromocriptine, inhibited 1,3,7-trimethyluric acid formation with Kis of 0.75 microM and 5 microM, respectively, thus further supporting the involvement of CYP3A4 in the 8-hydroxylation of caffeine. The study shows that fluvoxamine, as expected, is a potent inhibitor of the metabolism of caffeine in vitro. PMID- 9868742 TI - Effects of tamoxifen, toremifene and chloroquine on the lysosomal enzymes in cultured retinal pigment epithelial cells. AB - Retinal pigment epithelial cells carry out phagocytosis and digestion of material shed from the photoreceptor outer segments. In this process, the integrity of lysosomal enzymes is of major importance. In the present study the effects of tamoxifen, toremifene and chloroquine on the activity of two lysosomal enzymes (cathepsin D and N-acetyl-beta-D-glucosaminidase) in the retinal pigment epithelial cells were studied. Retinal pigment epithelial cells from pig eyes were cultured for two weeks in Dulbecco's Modified Eagle Medium, after which the cells were exposed to 1-40 microM concentrations of tamoxifen citrate, toremifene citrate and chloroquine diphosphate. To eliminate possible medium-borne oestrogenic mechanisms, the test was repeated using phenol red-free medium with charcoal-stripped fetal calf serum. The exposure time was one week, after which the lysosomal enzymes cathepsin D and N-acetyl-beta-glucosaminidase were determined. Cellular injuries were assessed by quantifying the leakage of lactate dehydrogenase into the culture medium. Cathepsin D and N-acetyl-beta-D glucosaminidase showed different sensitivities to tamoxifen, toremifene and chloroquine. The main lysosomal protease cathepsin D was more sensitive than N acetyl-beta-D-glucosaminidase to the effects of tamoxifen and toremifene, possibly due to their antioestrogenic properties. The phenol red-free medium with charcoal-stripped serum seemed to make the drugs more effective than the reference medium. Chloroquine had only a minor effect on the lysosomal protease cathepsin D, but a clearer effect could be seen on N-acetyl-beta-glucosaminidase. PMID- 9868743 TI - Aspects on tail-flick, hot-plate and electrical stimulation tests for morphine antinociception. AB - The objective of this study was to compare the results of three nociceptive tests, tail-flick, hot-plate and electrical stimulation vocalisation, reflecting the responses from different sites in the CNS. A subcutaneous morphine dose (5 mg/kg) was administered to three parallel groups of rats in which the nociceptive response was measured by one of the three methods. The baseline decreased during the period of measurement for the hot-plate test, but remained stable for the other methods. The spinally mediated tail-flick response was more sensitive to the morphine effects as compared to the supraspinally mediated hot-plate and electrical stimulation vocalisation responses. The electrical stimulation vocalisation-test demonstrated more even effect-time profiles and less variability among the rats than did the tail-flick and the hot-plate methods. In the tail-flick group, 59% of the observations attained the cut-off latency at this morphine dose, leading to underestimation of the peak effect, the area under the effect curve (AUEC), and the variability among the rats. In the hot-plate group, 13% of the observations were at the cut-off latency, and 2% in the electrical stimulation vocalisation group. Different ways of presenting the data are discussed. In conclusion, the test selected for measuring the nociceptive response will influence the effect-time profile and subsequently any pharmacodynamic parameters describing it. PMID- 9868744 TI - Pharmacokinetics and antidiuretic effect of intravenous administration of desmopressin in orally overhydrated male volunteers. AB - In order to study the antidiuretic activity of desmopressin in healthy human subjects, a study design has previously been used with subjects orally hydrated. The objective of the present study was to investigate the pharmacokinetics and the antidiuretic activity of desmopressin administered as an intravenous infusion to eight orally hydrated male volunteers. After initial ingestion of water corresponding to 15 ml.kg-1 body weight, the overhydration was maintained by replacing the urinary fluid loss by drinking-water. Desmopressin (4 micrograms) was administered intravenously over 10 min. 1.5 hr after the start of the hydration procedure. Blood was sampled and urine was collected at intervals throughout the study day (9.5 hr). The terminal half-life of elimination (t1/2,) of desmopressin was calculated to be 2.97 +/- 0.24 (S.E.M.) hr while the clearance was 1.77 +/- 0.10 ml.min.-1.kg-1 and the volume of distribution at steady state was 373 +/- 30 ml.kg-1. The infusion caused a marked and long lasting reduction of the urine flow rate. Four hr after the start of the infusion the mean urine osmolality was 909 +/- 46 mOsm.kg-1. PMID- 9868746 TI - A theoretical approach to the Ferguson principle and its use with non-reactive and reactive airborne chemicals. AB - The Ferguson principle has been widely used in toxicology to separate or indicate possible mechanisms for acute toxic effects of chemicals. However, this principle has never been adequately tested because of the lack of a database containing a sufficient number of both types of chemicals, non-reactive and reactive, that the Ferguson principle purports to separate. Such a database is now available. In this report a theoretical framework for the Ferguson principle is presented, regarding one of the acute toxicological effects of volatile airborne chemicals: sensory irritation. Previously obtained results on series of non-reactive and reactive chemicals are then used to demonstrate that the Ferguson principle can be extended to reactive chemicals by adding chemical reactivity descriptors to the physicochemical descriptors required by the Ferguson principle. This approach can be successful, provided that specific chemical reactivity mechanisms can be identified for the reactive chemicals of concern. The findings suggest that it is possible to replace the empirical Ferguson principle by formal mechanistic equations which will provide a better foundation for the understanding of the mechanisms by which airborne sensory irritants exert their action. PMID- 9868745 TI - Fluoroquinolone-induced motor changes in the guinea-pig isolated ileum. AB - The effects of norfloxacin and enoxacin were examined on spontaneous motor activity in the guinea-pig isolated ileum. Micromolar concentrations of both compounds caused a biphasic response consisting of relaxation followed by transient contraction. Relaxation to norfloxacin, which was unaffected by phentolamine, propranolol and hyoscine (each at 1 microM), was partially sensitive to tetrodotoxin (1 microM). This indicates that the response is partly mediated by non-adrenergic non-cholinergic (NANC) inhibitory nerves, and partly related to a direct action on the smooth muscle. Apamin (0.1 microM) and suramin (300 microM) inhibited norfloxacin-induced relaxations to an extent similar to that of tetrodotoxin. Conversely, NG-nitro-L-arginine (300 microM) was ineffective. In the presence of theophylline (100 microM) and 3-isobutyl-1 methylxanthine (10 microM), norfloxacin caused relaxation less effective than when added alone. Based on this observation, the NANC component of the relaxation apparently depends on ATP release, whereas the direct component might be due, at least in part, to phosphodiesterase inhibition. Norfloxacin-induced contractions were neurogenic and cholinergic in nature. They were reduced by indomethacin or S ketoprofen (both at 0.01-1 microM) and suramin (300 microM), suggesting involvement of local prostaglandin production probably induced by ATP release. Previous findings revealed that norfloxacin acted as a non-competitive antagonist at enteric GABAA receptors. In this study the same property was shared by enoxacin against the contractile response to 3-aminopropane sulphonic acid (3 APS), a GABAA receptor agonist. In conclusion, fluoroquinolones exert inhibitory and excitatory effects in the guinea-pig ileum. These are mediated by ATP, prostaglandin and acetylcholine release that might underlie, at least in part, the alterations of gastrointestinal motility observed after fluoroquinolone administration. Furthermore, isolated intestinal preparations might be useful to predict the GABAA-antagonist potential of this class of compounds. PMID- 9868747 TI - Sickness absence: a review of performed studies with focused on levels of exposures and theories utilized. AB - BACKGROUND: Despite the major impact sickness absence has on society, companies and individuals, surprisingly little scientific knowledge has been accumulated, and the studies that have been performed vary greatly. AIM AND METHOD: Examination of about 320 studies of sickness absence regarding structural levels of exposures studied and theories utilized. RESULTS: Theories concerning sickness absence were found to vary greatly. Along with exposures and factors explaining sickness absence these theories were categorized with respect to different structural levels, i.e. they were deemed national, occupational, or individual. A classification of different types of absences is presented. CONCLUSION: Although most of the reviewed studies were performed within the realm of medicine, only a few of the investigators used medical explanatory models or even considered the health status of individuals. Moreover, it is known that factors at "higher" structural levels have a substantial effect on sickness absence, but these were seldom taken into account, or even mentioned, in the reports scrutinized. PMID- 9868748 TI - Short form 36 (SF-36) health survey: normative data from the general Norwegian population. AB - Anchoring health-related quality of life (HRQOL) measures in population norms makes clinical interpretations more meaningful and is in accordance with practice in other fields of medicine. In this paper norms for the Short Form 36 (SF-36) are presented in a random sample, representative of the general Norwegian population. In addition, sociodemographic variables affecting the scale scores are explored and discussed. The response rate was 67%, being lowest among subjects aged 70 years or over. Data-completeness strongly declined with increasing age. Physical health scales were also strongly affected by age. In all scales, with the exception of general health perceptions, women reported having poorer health than men. Marital status affected the four mental health scales. Educational status affected all the scales, but the effect was smallest in the mental scales. These norms can be employed for comparison in case-control studies, or to interpret HRQOL changes in prospective studies. Differences in social status should be given special attention. Caution should be exercised when assessing subjective health or employing the norms among subjects aged 70 years or over. PMID- 9868749 TI - Equity in the delivery of health care in Sweden. AB - There is mutual agreement that health care should be delivered according to need. In this article, although we employ different specifications for need, we conclude that there is inequity in the delivery of health care in Sweden. Higher income groups visit doctors more often than lower income groups in relation to need, but lower income groups remain in hospital longer once they have been admitted. These findings may be interpreted to mean that lower income groups, for various reasons, wait too long before visiting a doctor for a specific disease, the consequence being that the disease is so serious by the time the doctor is contacted that inpatient care and a longer time in hospital are necessary. The policy implication of this situation is that lower patient fees and/or higher incomes may help to reduce the inequities in health care. PMID- 9868750 TI - Impact of user charges and socio-economic environment on visits to paediatric trauma unit in Finland. AB - In this report we (i) measure the strength of the association between paediatric trauma visit rates and regional socioeconomic and demographic variables, and (ii) quantify any selective impact of user charges on service demand in socioeconomically and demographically different areas. During the period 1989-94, a total of 30,362 home and leisure injury visits were made to the Aurora City Hospital. The visit rates are analysed using a random effects model. In addition, the areas are ranked into three groups in accordance with a socioeconomic index measure, and the annual visit rates of the three groups are calculated. We found fairly minor differences in children's visit rates between unequal socioeconomic areas, and it is apparent that socioeconomic status cannot explain the change in visit rates as a result of the introduction of user charges. We speculate that these changes in emergency visit rates will not lead to long-term health problems among any segment of the city population. PMID- 9868751 TI - Finnish health centre physicians' participation in family planning. PMID- 9868752 TI - Socioeconomic differences in health indices, social networks and mortality among Swedish men. A study of men born in 1933. AB - BACKGROUND: In previous survey we found large socioeconomic differences in mortality among urban Swedish men which remained unexplained after controlling for smoking and standard coronary risk factors. The present analysis was undertaken in order to investigate a broader set of possible explanatory factors in another cohort of Swedish men. STUDY POPULATION AND METHODS: Occupation was coded into five occupational classes for 717 of 776 participant men from a random population sample of 1016 men who were born in 1933. All were living in Goteborg and were 50 years old at the baseline examination. After 12 years' follow-up, 68 of the 717 men had died (9.5%). RESULTS: Low occupational class was associated with a higher prevalence of smoking at baseline, but no association was found with systolic blood pressure, body mass index, waist to hip ratio, serum triglycerides or serum cholesterol. Subjects from higher socioeconomic strata were taller, had higher maximum peak respiratory flow, lower plasma fibrinogen and lower body temperature. Low occupational class was associated with low social integration, low home activity levels, low levels of activity outside home and low social activity levels (p = 0.001 for all) and with low emotional support (p = 0.018). There were also associations between low occupational class and poor self-perceived health, as well as with several cardiovascular symptoms. During 12 years' follow-up, there was a graded and inverse relationship between occupational class and mortality from all causes. The highest mortality was found among the men who could not be classified (23 per 1,000 person years) Of the men in the lowest occupational class, 12 per 1,000 died, compared to 3 per 1,000 in the highest class (relative risk 3.7 (1.4-9.8)). After controlling for smoking, the relative risk decreased to 3.2 (1.2-8.6) and after further adjustment for emotional support, self-perceived health, activity level at home, and peak expiratory flow, the relative risk was still twofold but not significantly so (RR 2.1 (0.8-5.8)). CONCLUSION: We were able to confirm earlier results as to the wide mortality differentials in urban middle-aged men in Sweden. There were also large differences in several other factors, including constitutional factors, health variables, lifestyle and social support indices, which explained important parts of the social mortality gradient, the most prominent being smoking, respiratory function, social network factors and subjective health. PMID- 9868753 TI - Socioeconomic correlates of infant mortality in Hong Kong, 1979-93. AB - Although Hong Kong's infant mortality is among the lowest in the world, there may still be subgroups in the population with unusually high and possibly avoidable mortality rates. We conducted an ecological study on the relationship between socioeconomic deprivation and infant mortality in Hong Kong by using government data from three periods: 1979-83, 1984-88 and 1989-93. The study population comprised all infant births in 65 modified districts in Hong Kong in the period 1979-93. Infant, neonatal and post-neonatal mortality rates (IMRs, NMRs and PNMRs) were used as the health indicators. An F score was derived from highly correlated socioeconomic variables by factor analysis and used as a summary index of socioeconomic status. In 1979-83, socioeconomic deprivation was found to be significantly associated with high IMRs and high NMRs in both sexes, while in 1984-88 this association was observed only in baby girls. None of the observed associations were significant in 1989-93. Overall, the territory's infant mortality rates fell from 10.2 per thousand live births in 1979-83 to 5.6 per thousand live births in 1989-93. Individual-based studies are needed to ascertain whether this apparent disappearance of the socioeconomic relationship with infant and neonatal mortality is real. PMID- 9868754 TI - Social differences in health in an affluent Danish county. AB - The aim of the present study was to examine whether selected health indicators were related to the level of social conditions in the municipalities in an affluent Danish county with a tax system aiming at narrowing income differences and with the same free health facilities for all. The 18 municipalities were separated into three groups by a sum score calculated on the basis of 10 social status variables. For each of the three groups average mortality, hospital discharge rates, cancer incidence and health services use were calculated with data from National Registers. There was a distinct gradient in all-cause, infant and cause-specific mortality, cancer incidence and use of health services related to level of social conditions. Apart from the incidence of breast cancer and melanoma, and use of general practice, all the analysed health indicators were most frequent in the group of the least affluent municipalities. It is concluded that even in an affluent county, the variation in the socioeconomic characteristics of neighbouring municipalities could be related to differences in health. PMID- 9868755 TI - Trends in smoking prevalence in Danish adults, 1964-1994. The influence of gender, age, and education. AB - BACKGROUND: Studies of time trends in smoking prevalence provide a better understanding of the determinants of smoking. The present study analyses changes over time in the prevalence of smoking and heavy smoking in relation to sex, age, and education. METHODS: Data on smoking behaviour were collected by questionnaire in random samples of the general population in the area of Copenhagen. The database used included 71,842 measurements of smoking behaviour for 32,156 subjects aged 30 years or more, who had been examined at intervals between 1964 and 1994. In bi- and multivariate analyses the effects of sex, age, education, time period, and study group on the prevalence of smoking and of heavy smoking were assessed. RESULTS: Smoking was least prevalent in women, in the oldest age group (more than 70 years), and among those with 8 years or more of school education. During the study period (from 1964/74 to 1990/94), the prevalence of smoking decreased from 72% to 54% in men and from 52% to 46% in women. In both men and women this decrease was smallest in the least educated (less than 8 years of school education). Heavy smoking was also least prevalent in women, in the oldest age group, and among the well educated. During the study period, the unadjusted prevalence of heavy smoking decreased from 52% to 38% in men, while it increased from 17% to 21% in women. The multivariate analysis showed that the time trend for heavy smoking only depended on sex, while educational attainment and age had no impact on the trend. CONCLUSION: During the last 30 years the prevalence of smoking has decreased in Denmark. The decrease has been smallest in women, and among the least educated, and the increasing trend in the prevalence of heavy smoking in women is a cause for concern. PMID- 9868756 TI - Tobacco consumption and national rates of personal violence (suicide and homicide) PMID- 9868757 TI - Risk factors of cerebral palsy. PMID- 9868758 TI - Epidemiological observations in a Corriedale flock affected by Brucella ovis. AB - Brucellosis in sheep, caused by Brucella ovis, is primarily a chronic infectious disease of rams with epididymitis as its most characteristic lesion. Six hundred rams from an infected farm were clinically and serologically examined once a year, over a 3-year period. An increase from 2.1% to 6.3% in the prevalence of animals serologically positive to B. ovis occurred over the 3 years. However, the prevalence of rams with lesions in the reproductive tract declined from 14.2% to 6.5% in the third year following one year of strict culling of clinically affected and rams that were serologically positive for B. ovis. Clinical lesions found in the 179 affected rams fell into two main categories: rams with epididymitis and rams with affected lymph nodes. These results suggest that the prevalence of the disease relates mainly to the sexual activity of the animal and not to age in itself. A single cull based on the results of clinical examination and serological test results was unable to decrease the prevalence of B. ovis in an extensive Corriedale sheep flock. PMID- 9868759 TI - Adaptive acquisition of novobiocin resistance in Pasteurella multocida strains of avian origin. AB - Naturally occurring strains of Pasteurella multocida are atypically susceptible to hydrophobic antibiotics such as novobiocin, despite their Gram-negative cell envelope ultrastructure. Four strains adaptively resistant to 1000 micrograms/ml of novobiocin were obtained by sequentially subculturing cell surface hydrophobic variants of avian origin in the presence of increasing antibiotic concentrations. Adaptive novobiocin resistance was accompanied in all cases by the concomitant acquisition of resistance to coumermycin, a hydrophobic antibiotic possessing the same mechanism of action, but not to the functionally disparate hydrophobic antibiotic rifamycin. The acquisition of resistance was not accompanied by alterations in the lipid composition of the cell envelope. Subsequent growth of adaptively resistant strains in the absence of novobiocin did not result in the restoration of susceptibility to either novobiocin or coumermycin. Acquisition of adaptive resistance in encapsulated parental strains resulted in an inability to synthesize capsular material and enhanced cell surface hydrophobicity; however, parental encapsulation and decreased cell surface hydrophobicity were restored upon removal of novobiocin. These data suggest that acquisition of adaptive resistance to novobiocin conferred in this manner is the result of a stable genetic event affecting the mechanistic target of both novobiocin and coumermycin rather than a physiological adaptation involving outer membrane impermeability. PMID- 9868760 TI - A possible correlation between the blood leukocyte formula and the use of glucocorticoids as growth promoters in beef cattle. AB - Clinical analyses designed to set welfare parameters were performed on blood drawn from the caudal vein of 14 groups of cattle (young bulls and heifers) (n = 10) from 480 to 550 kg b.w., each group representative of a different farm. The leukocyte formula exhibited a lymphocytopenia in four groups compared with the values from a control group (n = 50). This finding was related to the possible illicit use of corticoids as growth promoters in meat production. The individual plasmas tested negative by two different ELISA kits for corticosteroids, but chemical analyses by LC-MS/APCI (detection limit 0.5 ng/ml) on the pooled plasma of each of the 14 groups revealed the presence of beclomethasone and fluocinolone acetonide in 3 of the 4 suspect farms. These corticosteroids are not always efficiently screened by commercially available immunoassays. The epidemiological reliability of blood analysis as a screening test for such drugs is discussed in the light of the need for quality certification of the whole meat production processes 'from farm to fork', and for enhanced animal welfare. PMID- 9868761 TI - The pharmacokinetics and efficacy of long-term low-level and split-dose administration of albendazole through in-feed formulations against ovine and caprine parasitic gastroenteritis. AB - Two trials were conducted against natural and experimentally induced parasitic gastroenteritis in sheep and goats using an in-feed formulation of albendazole to evaluate its therapeutic and prophylactic efficacy. In the first trial, albendazole was incorporated in feed pellets to deliver an average daily dose of 0.7 mg/kg body weight in order to evaluate its prophylactic efficacy. In the second trial, feed pellets were offered to deliver an average total dose of 8.0 mg/kg body weight in two equal split doses in order to evaluate its curative efficacy. Sustained plasma concentrations of the active compound, albendazole sulphoxide, and its metabolite albendazole sulphone, sufficient to prevent establishment of infection, were achieved when the animals were allowed to feed on medicated pellets for 10 consecutive days. The bioavailability of the metabolites of albendazole following the administration of a therapeutic dose in two split doses of the in-feed formulation was sufficient to remove established adult nematodes. The concentrate feed pellets could be used for self-medicating small ruminants for therapeutic use as well as for prophylaxis based on their strategic use appropriate to the epidemiology of the parasitic disease. PMID- 9868762 TI - Plasma, urinary and biliary residues in cattle following intramuscular injection of nortestosterone laurate. AB - The synthetic androgen 19-nortestosterone (beta-NT) has been used illegally as a growth promoter in cattle production in the European Union. Elimination of beta NT and its metabolites in plasma, urine and bile was studied in three cattle with cannulated gallbladders following intramuscular injection at a single site of 500 mg of the laurate ester (NTL) containing 300.5 mg beta-NT. Using enzyme immunoassay quantification, plasma Cmax of free beta-NT was 0.5 +/- 0.15 microgram/L (mean +/- SEM). Concentrations of free beta-NT in plasma were consistently greater than the assay limit of quantification (0.12 microgram/L) for 32.7 +/- 13.42 days. Mean residence time for the beta-NT in plasma was 68.5 +/- 20.75 days. Following sample preparation by immunoaffinity chromatography, high-resolution GC-MS was used to quantify beta-NT and alpha-NT in urine and bile. beta-NT was detected irregularly in urine from two of the three animals post injection. The principal metabolite present in the urine, alpha-NT, was detected for 160.3 +/- 22.67 days post injection. Cmax for alpha-NT in urine was 13.7 +/- 5.14 micrograms/L. Mean urinary AUC0-183 days for alpha-NT was 845.7 +/- 400.90 (microgram h)/L. In bile, alpha-NT was the only metabolite detected for 174.3 +/- 8.67 days post treatment. Cmax for alpha-NT in bile was 40.8 +/- 12.70 micrograms/L and mean biliary AUC0-183 days for alpha-NT was 1982.6 +/- 373.81 (microgram h)/L. Concentrations of alpha-NT in bile samples were greater than those in urine samples taken at the same time. The mean ratio of biliary:urinary AUC0-183 days was 3.0 +/- 0.72. It is concluded that bile is a superior fluid for detection of alpha-NT following injection of NTL, owing to the longer period during which residues may be detected after administration. PMID- 9868764 TI - Similarity in moth-fly specific larvicidal activity between two serologically unrelated Bacillus thuringiensis strains. AB - Parasporal inclusions of a Bacillus thuringiensis isolate designated 92-KU-105-9 (H14/19) exhibited unusual larvicidal activity, specific for the moth-fly, Telmatoscopus albipunctatus (Diptera: Psychodidae), similar to that of a previously reported B. thuringiensis serovar leesis (H33) strain. The LC50 value of the purified inclusions was 4.92 micrograms ml-1 for the moth-fly larvae, while no mortality was shown in the mosquitoes Culex pipiens molestus and Anopheles stephensi, at protein concentrations up to 10 mg ml-1. Morphologically, the inclusion was a homogeneous globular body surrounded by an electron-dense, thick envelope. Multilamellar inner structure was evident between envelope membrane and inclusion matrix. SDS-PAGE revealed that the inclusions consist of five proteins with molecular masses of 72, 70, 68, 56 and 30 kDa. These proteins cross-reacted with the antibodies against inclusion proteins of the serovar leesis strain. High homologies existed in N-terminal amino acid sequences between the three major proteins (72, 70 and 68 kDa) and the two established protein classes, Cry4A and Cry10A. PMID- 9868763 TI - Responses of guinea-pig lung parenchymal strips to tracheobronchial lavage fluid from horses affected with summer pasture-associated obstructive pulmonary disease. AB - The response of parenchymal strips from guinea-pig lungs to tracheobronchial lavage fluid (TBLF) collected from 8 normal horses and from 8 affected with summer pasture-associated obstructive pulmonary disease (SPAOPD) was determined. TBLF was collected during the summer (July) and winter (February) seasons. The serum/TBLF urea nitrogen ratio was used to standardize the mediator concentration in the TBLF. Four strips were used from each guinea-pig. The first strip did not receive any antagonist and served as the control. The second, third and fourth strips received antagonists of PGE2, LTD4 and PAF, respectively at 10(-6) mol/L for 30 min. The tissues were then precontracted with a dose of histamine (10(-5) mol/L) and their responses to 1 ml of TBLF were determined. The study showed that TBLF obtained in the summer from unaffected horses produced a significantly greater relaxation than that from the affected horses, whereas TBLF obtained in the winter from unaffected or affected horses did not cause a significantly different degree of relaxation. Among the antagonist-treated strips, only those exposed to the PGE2 blocker showed a significant reduction in the relaxation caused by TBLF obtained in the summer from SPAOPD horses. This suggests that PGE2 is an important mediator present in the summer in the TBLF from horses affected with SPAOPD. PMID- 9868765 TI - Isolation and characterization of toluene-sensitive mutants from Pseudomonas putida IH-2000. AB - Two toluene-sensitive mutants were generated from Pseudomonas putida IH-2000, the first known toluene-tolerant isolate, by Tn5 transposon mutagenesis. These mutants were unable to grow in the presence of toluene (log P(ow) 2.8) but they could grow in medium overlaid with organic solvents having a log P(ow) value higher than that of toluene such as p-xylene (log P(ow) 3.1), cyclohexane (log P(ow) 3.4) and n-hexane (log P(ow) 3.9). The Tn5 transposable element knocked out a cyoB-like gene in one mutant and a cyoC-like gene in the other mutant. Seven open reading frames were found in a 5.5-kb region containing the cyoB- and cyoC like genes of strain IH-2000. ORFs 3.7 showed significant identity to the cyoABCDE gene products of Escherichia coli, but ORFs 1 and 2 showed no significant homology to any protein reported so far. The growth patterns of the Tn5 mutants with the inactivated cyo-like gene were similar to that of the wild type strain in the absence of organic solvents, although the doubling times were slightly longer than that of the wild-type strain. Our findings indicate that cyo is an important gene for toluene tolerance, although its role is still unclear. PMID- 9868766 TI - Feasibility of using prokaryote biosensors to assess acute toxicity of polycyclic aromatic hydrocarbons. AB - The aim of this study was to assess the acute toxicity of polycyclic aromatic hydrocarbons using lux-marked bacterial biosensors. Standard solutions of phenanthrene, pyrene and benzo[a]pyrene were produced using 50 mM hydroxpropyl beta-cyclodextrin solution which contained each respective polycyclic aromatic hydrocarbon at 6.25 times the aqueous solubility limit of the compound. The polycyclic aromatic hydrocarbon solutions were incubated with each of the biosensors for 280 min and the bioluminescence monitored every 20 min. Over the incubation time period, there was no significant decrease in bioluminescence in any of the biosensors tested with the exception of Rhizobium leguminosarum biovar trifolii TA1 luxAB. In this series of incubations, there was a dramatic increase in bioluminescence in the presence of phenanthrene (2.5 times) and benzo[a]pyrene (3 times) above that of the background control (biosensor without polycyclic aromatic hydrocarbon) after 20 min. Over the next 3 h, bioluminescence decreased to that of the control. An ATP assay was carried out on the biosensors to assess if uncoupling of the oxidative phosphorylation mechanisms in the respiratory chain of the cells had occurred. However, it was found that the polycyclic aromatic hydrocarbons had no effect on the organisms indicating that there was no uncoupling. Additionally, mineralisation studies using 14C-labelled polycyclic aromatic hydrocarbons showed that the biosensors could not mineralise the compounds. This study has shown that the three polycyclic aromatic hydrocarbons tested are not acutely toxic to the prokaryotic biosensors tested, although acute toxicity has been shown in other bioassays. These results question the rationale for using prokaryote biosensors to assess the toxicity of hydrophobic chemicals, such as polycyclic aromatic hydrocarbons. PMID- 9868767 TI - Characterisation of CMY-4, an AmpC-type plasmid-mediated beta-lactamase in a Tunisian clinical isolate of Proteus mirabilis. AB - A strain of Proteus mirabilis resistant to beta-lactams, including cefoxitin, was isolated from the urine of a woman from Tunisia. Its antibiotic susceptibility pattern and that of the Escherichia coli transconjugant suggested the presence of an AmpC-type beta-lactamase. Two bands of beta-lactamase activity (pI 5.4 and 9.2) were detected by isoelectric focusing. The nucleotide sequence of the gene encoding the AmpC-type enzyme was determined. The deduced amino acid sequence was 98-99% identical to CMY-3 and to those of the plasmid-mediated AmpC-type beta lactamases originated from Citrobacter freundii and 97% identical to the chromosome-encoded beta-lactamase of a Tunisian clinical isolate of C. freundii. This enzyme differs from CMY-2 by one substitution (Arg for Trp at position 221) and from CMY-3 by two substitutions (Glu for Gly at position 42 and Ser for Asn at position 363) and we propose the denomination CMY-4. PMID- 9868768 TI - Mutational analysis of exopolysaccharide biosynthesis by Lactobacillus sakei 0-1. AB - Lactobacillus sakei strain 0-1 produces an exopolysaccharide (EPS) consisting of glucose and rhamnose in a ratio of 3:2. As part of a biochemical and molecular analysis of the EPS biosynthetic pathway in L. sakei strain 0-1, we have isolated a random set of EPS-negative mutants. Following treatment of cells with the mutagen ethylmethane sulfonic acid, a total of 10 mutants were identified that lacked the clear ropy appearance of wild-type colonies on agar plates. Their characterization revealed that eight mutants had completely lost the ability to synthesize the normal EPS. Six of these mutants lacked activities of enzymes involved in the biosynthesis of dTDP-rhamnose, required for EPS production. Only mutant strains 12 and 20 were directly affected in EPS synthesis. Strain 12 synthesized EPS with a different sugar composition, however. Interestingly, strain 12 showed temperature-dependent EPS synthesis, with the highest amounts synthesized at 12 degrees C, and low amounts at the optimal temperature for growth (30 degrees C). Two mutants were in fact EPS-positive, producing the normal EPS, but displayed a different cell morphology (elongated cells), indicating a modification in cell wall synthesis. PMID- 9868769 TI - Effect of temperature and viscosity on the motility of the spirochete Treponema denticola. AB - Treponema denticola is an oral spirochete associated with periodontal diseases. Because bacterial motility is likely to be a potential virulence factor, we investigated the effect of viscosity and temperature on cell speed. In agreement with the work of others, translational motility was a function of the macroscopic viscosity of the medium. In addition, we found that although the speed of spirochetes was slow at 25 degrees C (4 microns s-1), it increased quite markedly at 35 degrees C (19 microns s-1). The results indicate that both viscosity and temperature are critical factors in T. denticola translational motility. PMID- 9868770 TI - Construction and physiological analysis of a Xanthomonas oryzae pv. oryzae recA mutant. AB - A Xoo recA insertion inactivation mutant was constructed. The mutant, lacking RecA, showed increased sensitivity towards mutagen killing. This phenotype could be complemented by a cloned, functional recA. Unlike other bacteria, both the recA mutant and the parental strain had similar level of resistance to H2O2 killing and peroxide-induced mutagenesis. PMID- 9868771 TI - Hydration of linoleic acid by bacteria isolated from ruminants. AB - Two strains of Enterococcus faecalis isolated from the ovine rumen and known to hydrate oleic acid were shown to transform linoleic acid by hydration into two products. The products, identified as 10-hydroxy-12-octadecenoic acid and 13 hydroxy-9-octadecenoic acid, were formed during stationary phase in yields of 13% and 6% respectively. Yields increased to 22% and 14% when culture conditions were optimised. To our knowledge, this is the first report of 13-hydroxy-9 octadecenoic acid production by bacteria. During a search for further linoleic acid-hydrating bacteria, a strain of Streptococcus bovis isolated from bovine faeces and the ruminal strain S. bovis JB1 were found to hydrate linoleic acid. Both strains formed only one product and the most rapid appearance occurred during exponential growth. The S. bovis product, identified as 13-hydroxy-9 octadecenoic acid, formed in a yield of 28%. This study provides the first information on linoleic acid hydration by ruminal bacteria. PMID- 9868773 TI - Rhizobium leguminosarum bv. viciae hypA gene is specifically expressed in pea (Pisum sativum) bacteroids and required for hydrogenase activity and processing. AB - Rhizobium leguminosarum bv. viciae strain UPM791 induces in symbiosis with peas the synthesis of a nickel-containing hydrogenase which recycles the hydrogen evolved by nitrogenase. The genes required for synthesis of this hydrogenase, hupSLCDEFGHIJKhypABFCDEX, are clustered in the symbiotic plasmid. Analysis of a hypA-deficient mutant showed that HypA is essential for symbiotic hydrogenase activity and required for correct processing of the hydrogenase large subunit. Unlike other microoxically induced hyp genes, the hypA gene was only expressed in pea bacteroids from its own promoter. The hypA mRNA 5' end was mapped 109 bp upstream of the translational start codon. This distinct pattern of expression suggests a different role for HypA and the remaining Hyp proteins in hydrogenase synthesis. PMID- 9868772 TI - Effect of substitution of Asn for Arg-276 in the cefotaxime-hydrolyzing class A beta-lactamase CTX-M-4. AB - The effect of substitution of asparagine for arginine at position 276 (Ambler's numbering) on the properties of the extended-spectrum beta-lactamase CTX-M-4 was studied. Compared with CTX-M-4, the mutant beta-lactamase CTX-M-4(R276N) conferred lower levels of resistance to cefotaxime, ceftriaxone and aztreonam while the levels of resistance to penicillins and penicillin-inhibitor combinations were similar. Arg-276-->Asn substitution rendered CTX-M-4 slightly less susceptible to inhibition by clavulanate and tazobactam. It also caused a three-fold reduction in the relative rate of hydrolysis of cefotaxime. These results indicate that Arg-276 in CTX-M-type beta-lactamases may be implicated in hydrolysis of oxyimino-beta-lactams; they do not, however, support the hypothesis that Arg-276 is the functional equivalent of Arg-244 found in other class A beta lactamases. PMID- 9868774 TI - Transcript of a homolog of aflR, a regulatory gene for aflatoxin synthesis in Aspergillus parasiticus, was not detected in Aspergillus oryzae strains. AB - Some strains of Aspergillus oryzae were shown to have homologs of aflR, a regulatory gene for aflatoxin synthesis in Aspergillus parasiticus. Transcription of an aflR homolog was examined in six strains of A. oryzae having the homologs, using polymerase chain reaction (PCR) coupled with reverse transcription. No PCR product was obtained when the RNA prepared from the A. oryzae strains cultivated under aflatoxin-producing condition was used as template for amplification of the aflR cDNA. By contrast, a PCR product of the expected size was obtained with RNA from A. parasiticus NIAH-26 processed by the same procedure. From genomic DNA of these strains, PCR products of the same size as above were obtained. Possible degradation of the aflR mRNA in the RNA preparation of the A. oryzae strains was negligible, because the calmodulin transcript was detected by PCR from the same RNA samples. Thus, the aflR homologs in the non-aflatoxigenic A. oryzae strains examined are not expressed even under aflatoxin-producing condition. PMID- 9868776 TI - Isolation of myxobacteria from the marine environment. AB - In an attempt to isolate indigenous marine myxobacteria from coastal samples, we obtained two swarm forming bacteria. Both isolates formed cell aggregates which, at least in one isolate, developed to fruiting body-like structures consisting of a mass of myxospore-like cells. The optimum NaCl concentrations for their growth were between 2 and 3%, comparable to the NaCl concentration of seawater. This growth characteristic strongly suggests that the two isolates are specific marine bacteria. The 16S rDNA sequence studies indicated that the two isolates were related to the genus Nannocystis. Based on the phylogenetic distances between branches, we concluded that the isolates should be assigned to two new myxobacterial genera. PMID- 9868775 TI - Lactate causes changes in gonococci including increased lipopolysaccharide synthesis during short-term incubation in media containing glucose. AB - Gonococci (strain BS4(agar)), emerging from lag-phase during 1-1.5 h incubation in a medium containing glucose (28 mM) and either 5 microM or 50 microM sodium lactate, show enhanced capacity for their lipopolysaccharide (LPS) to be sialylated by cytidine 5'-monophospho-N-acetyl neuraminic acid. The sialyltransferase content of the lactate-treated gonococci was not greater than that of control organisms and showed no differences in LPS components. However, the total LPS content of the lactate-treated gonococci was 10-20% higher than that of control organisms, so lactate enhancement may be due to more sialyl receptors becoming available due to an overall stimulation of LPS synthesis. The protein and pentose contents of the lactate-treated gonococci were also higher than those of controls, indicating stimulation of protein synthesis and ribosome production. Electron microscopy showed hair-like external appendages on control but not on lactate-treated gonococci. The above growth conditions are unnatural. However, when concentrations of glucose and lactate were adjusted to values akin to those occurring in vivo (glucose 5 mM alone and with either 1 mM or 10 mM lactate), and gonococcal multiplication occurred during the short incubation period (1-1.5 h), lactate again induced greater contents of LPS, protein and pentose. A high content of LPS, which will contribute to pathogenicity, should be a constant feature of gonococci growing in human urogenital tissues, where lactate is ever present with glucose. PMID- 9868777 TI - Transcriptional analysis of the isiAB operon in salt-stressed cells of the cyanobacterium Synechocystis sp. PCC 6803. AB - Expression of the isiA and isiB genes was analysed in the cyanobacterium Synechocystis sp. PCC 6803 grown in high salt or in iron-deficient medium. The detection of a 2.3-knt transcript in Northern blot experiments indicated cotranscription of isiAB in an operon, which was confirmed by reverse transcriptase PCR. The abundance of a monocistronic 1.25-knt isiA-specific mRNA was about 10-fold higher than the dicistronic message. The isiAB-specific transcripts were most abundant in cells adapted to 342 mM NaCl and under iron deficiency. The promoter of the operon was mapped to 211 bp upstream of the translational start. A putative Fur binding site was detected immediately upstream of the GTG start codon. A preliminary transcript of about 0.2 knt was detected in cells grown in conditions in which the isiAB operon was not transcribed. This indicates that a repressor binds to the identified Fur binding site and thus inhibits isiAB transcription under low salt and iron replete conditions. PMID- 9868778 TI - Characterization of non-membrane-damaging cytotoxin of non-toxigenic Vibrio cholerae O1 and its relevance to disease. AB - The non-membrane-damaging cytotoxin which causes dramatic cell rounding of cultured HeLa cells was purified to homogeneity from a clinical strain (WO5) of non-toxigenic Vibrio cholerae O1 Inaba belonging to the E1 Tor biotype. The purified protein has a denatured molecular weight of 35 kDa and a native molecular weight of approximately 37 kDa indicating the monomeric nature of the protein. The 15 N-terminal amino acid sequence of non-membrane-damaging cytotoxin showed complete homology to the hemagglutinin protease previously purified and characterized from V. cholerae O1. Purified non-membrane-damaging cytotoxin from V. cholerae O1 was immunologically and biochemically identical to that previously purified from V. cholerae O26. Non-membrane-damaging cytotoxin was found to be enterotoxic in rabbit ileal loop assay inducing accumulation of non-hemorrhagic fluid at 100 micrograms and elicited a concentration dependent increase in short circuit current and tissue conductance of rabbit ileal mucosa mounted on Ussing chambers. A significant serum immunoglobulin G response against non-membrane damaging cytotoxin was elicited by patients infected with V. cholerae O139 but not with V. cholerae O1. These properties make non-membrane-damaging cytotoxin a potential virulence factor of V. cholerae which should be taken into consideration while making live, attenuated recombinant vaccine strains against cholera. PMID- 9868779 TI - Characterization of a large motility gene cluster containing the cheR, motAB genes of Listeria monocytogenes and evidence that PrfA downregulates motility genes. AB - Through the analysis of a non-motile mutant of Listeria monocytogenes, we identified and characterized a locus containing the cheR, motA and motB genes. These three genes are homologous to the cheR, and motA/B genes of Bacillus subtilis which in this organism are 954 kb apart. The gene organization in Listeria is also not similar either to that of Escherichia coli in which cheR and motAB are 5.9 kb apart. CheR and motA/B, as previously reported for flaA, the flagellin gene, are thermoregulated with a higher expression at 25 degrees C and low expression at 37 degrees C. In a delta prfA strain, motA expression was derepressed at 37 degrees C, suggesting that PrfA, the transcriptional activator of virulence genes, downregulates motility genes in Listeria at 37 degrees C. PMID- 9868780 TI - Internal size variations in Tn1546-like elements due to the presence of IS1216V. AB - In this study, internal size variations in the VanA gene cluster Tn1546, encoding resistance to glycopeptides, is described. Studies of previously uncharacterized size variations of an internal region, encoding the vanX and vanY genes of Tn1546, revealed that these variations were due to the presence of the IS sequence, IS1216V. This IS sequence has previously been found integrated in Tn1546. Integration of the IS1216V element created both deletions and a duplication in a non-essential region of Tn1546. In several isolates, the entire vanY gene was deleted, proving that this gene is non-essential for vancomycin resistance. PMID- 9868781 TI - Uptake and incorporation of choline and ethanolamine into lipoteichoic acid and teichoic acid by the choline-independent mutant JY2190 of Streptococcus pneumoniae. AB - Lipoteichoic acid- and teichoic acid-containing muropeptides were isolated from choline- or ethanolamine-grown cells of the choline-independent mutant JY2190 of Streptococcus pneumoniae. Choline was taken up and incorporated into lipoteichoic acid and teichoic acid with 81% efficiency, compared with the parent strain Rx1. With similar efficiency, ethanolamine was incorporated. Accordingly, the mutant is a valuable tool for identifying the individual genes encoding the enzymes of choline utilisation, because any of these genes can be deleted without affecting viability and growth rate. PMID- 9868782 TI - Lysine is synthesized through the alpha-aminoadipate pathway in Thermus thermophilus. AB - A 3.8-kb DNA fragment which was able to complement the mutation of a lysine auxotrophic Thermus thermophilus mutant was cloned from T. thermophilus HB27. Sequence analysis of the 3.8-kb fragment indicated the presence of three open reading frames including a truncated one. The predicted amino acid sequences of two of the three open reading frames showed 55.2% and 45.0% identity with homocitrate synthase and homoaconitate hydratase of Saccharomyces cerevisiae, respectively. These two enzymes act as lysine biosynthetic enzymes through the alpha-aminoadipate pathway which has been reported in S. cerevisiae and fungi. Each of the two open reading frames in T. thermophilus was disrupted by integration of the heat-stable kanamycin nucleotidyltransferase gene. The resulting mutants showed lysine auxotrophy, which could be complemented with alpha-aminoadipate but not with diaminopimelate. These results indicate that lysine was synthesized through the alpha-aminoadipate pathway and not through the diaminopimelate pathway in T. thermophilus. PMID- 9868783 TI - Expression, purification, reconstitution and inhibition of Ustilago maydis sterol 14 alpha-demethylase (CYP51; P450(14DM)). AB - Triadimenol and tebuconazole are potent inhibitors of the sterol 14 alpha demethylation reaction in fungi which is catalysed by CYP51, a haem-thiolate containing enzyme belonging to the cytochrome P450 monooxygenase superfamily. Using CYP51 from the phytopathogen Ustilago maydis, a comparison of the sensitivity of the fungal enzyme to triadimenol and tebuconazole has been carried out. U. maydis CYP51 was purified to homogeneity as determined by SDS-PAGE and specific haem content. Catalytic activity was investigated following reconstitution with its respective NADPH cytochrome P450 reductase and proposed endogenous substrate, 24-methylenedihydrolanosterol. Addition of the triadimenol and tebuconazole induced type II spectral changes in the enzyme, with saturation occurring at equimolar azole concentrations. Inhibition of reconstituted activities showed a one-to-one sensitivity of the fungal CYP51 as judged by IC50 values. The implications for fungicide mode of action and treatment are discussed. PMID- 9868784 TI - Small genes/gene-products in Escherichia coli K-12. AB - Forty-two protein spots of observed M(r) 6-15 kDa were resolved by two dimensional gel electrophoresis, stained by Coomassie blue and subjected to Edman microsequencing. All of the proteins could be related back to their encoding open reading frames, thereby vindicating the bioinformatic tools currently utilised in their identification. However, only 14/42 gene-products were expressed as annotated. Translation was confirmed for 14 open reading frames with no attributed function (EcoGene Y-entries), while N-terminal sequence allowed the start codon to be accurately annotated for the genes yigF, yccU, yqiC, ynfD, and yeeX. The methionine start codon was cleaved in 11 gene-products (AtpE, Hns, RpoZ, RplL, CspC, YccJ, YggX, YjgF, HimA, InfA, RpsQ) and a further five showed loss of a signal peptide (PspE, HdeB, HdeA, YnfD, YkfE). Internal (Tig, AtpA, TufA) and N-terminal fragmentation (CspD, RpsF, AtcU) of much larger proteins was also detected, which may have resulted from physiological or translational processes. M(r) and pI isoforms were detected respectively for PtsH and GatB, each being phosphoproteins, as well as RplY which manifested differences with respect to predicted M(r) and pI. In addition, YjgF was shown to belong to a small gene family of unknown function with ancient conserved regions across procaryotes and eucaryotes. YgiN was revealed to have a paralogue and orthologues in Bacillus subtilis, Synechocystis sp., Mycobacterium tuberculosis, Neisseria gonorrhoea, and Rhodococcus erythropolis. Orthologues are also reported for YihD, YccU and YeeX. Of the 14 Y-genes, only YkfE possessed no detectable orthologues. These results highlight the need to complement genomic analysis with detailed proteomics in order to gain a better understanding of cellular molecular biology, while the confirmation of the open reading frame start codon using Edman degradation protein microsequencing has yet to be superseded by recent advances in mass spectrometry. PMID- 9868785 TI - Electroporation-mediated transformation of the ovine footrot pathogen Dichelobacter nodosus. AB - Studies on the potential virulence genes of the ovine footrot pathogen Dichelobacter nodosus have been hindered by the lack of a genetic system for this organism. In an attempt to accomplish the transformation of D. nodosus cells, we constructed a plasmid that contained part of a native D. nodosus plasmid and carried a tetracycline resistance gene that was located between the D. nodosus rrnA promoter and terminator. This plasmid was used to transform several D. nodosus strains to tetracycline resistance. Analysis of two independent transformants from each parental strain showed that in nearly all of these derivatives, the plasmid was not replicating independently, but that the tetracycline resistance gene had inserted by homologous recombination into one of the three rrn operons located on the chromosome. In most of the transformants, double reciprocal crossover events had occurred. These results are highly significant for genetic studies in D. nodosus and for footrot pathogenesis studies, since by using reverse genetics it will now be possible to examine the role of putative D. nodosus-encoded virulence genes in the disease process. PMID- 9868786 TI - An integron of class 1 is present on the plasmid pCG4 from gram-positive bacterium Corynebacterium glutamicum. AB - The streptomycin/spectinomycin resistance determinant of the 29-kb plasmid pCG4 from Corynebacterium glutamicum was found to be a part of a typical class 1 integron. The sequence analysis revealed that the integron (designated InCg) identified in this Gram-positive bacterium is almost identical to the integron InC present on the plasmid pSA1700 from the Gram-negative bacterium Pseudomonas aeruginosa. Differences in only two base pairs were found in the 3.8-kb sequence. One base substitution (G-->C) is present in the streptomycin/spectinomycin resistance determinant which is thus identical to the aadA2a gene from the integron In6 of the broad-host-range plasmid pSa. The other one (C-->G) is present in the extended -10 region of the integron promoter involved in expression of the antibiotic resistance gene. It was shown that this novel version of the integron promoter displays five times higher activity in both C. glutamicum and Escherichia coli than the original one. PMID- 9868787 TI - Identification and sequencing of a cDNA encoding 6-phosphogluconate dehydrogenase from a fungus, Cunninghamella elegans and expression of the gene in Escherichia coli. AB - The fungus, Cunninghamella elegans has been widely used in bioremediation and microbial models of mammalian studies in many laboratories. Using the polymerase chain reaction to randomly amplify the insert directly from the single non-blue plaques of a C. elegans cDNA library, then partly sequencing and comparing with GenBank sequences, we have identified a clone which contains C. elegans 6 phosphogluconate dehydrogenase gene. The polymerase chain reaction product was cloned into a plasmid, pGEM-T Easy vector for full insert DNA sequencing. The 6 phosphogluconate dehydrogenase gene (1458 bases) and the deduced protein sequence were determined from the insert DNA sequence. The gene was found by open reading frame analysis and confirmed by the alignment of the deduced protein sequence with other published 6-phosphogluconate dehydrogenase sequences. Several highly conserved regions were found for the 6-phosphogluconate dehydrogenase sequences. The 6-phosphogluconate dehydrogenase gene was subcloned and over-expressed in a plasmid-E. coli system (pQE30). The cell lysate of this clone has a very high 6 phosphogluconate dehydrogenase enzyme activity. Most of the recombinant protein in this system was formed as insoluble inclusion bodies, but soluble in high concentration of urea-buffer. Ni-NTA resin was used to purify the recombinant protein which showed 6-phosphogluconate dehydrogenase enzyme activity. The recombinant protein has a predicted molecular size correlating with that revealed by sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis. The C. elegans 6-phosphogluconate dehydrogenase was in a cluster with yeast' 6 phosphogluconate dehydrogenase in the phylogenetic tree. Bacterial 6 phosphogluconate dehydrogenase and higher organisms' 6-phosphogluconate dehydrogenase were found in different clusters. PMID- 9868788 TI - Co-regulation of lipoamide dehydrogenase and 2-oxoglutarate dehydrogenase synthesis in Escherichia coli: characterisation of an ArcA binding site in the lpd promoter. AB - The lipoamide dehydrogenase gene (lpdA) encoding the E3 subunits of both the pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase complexes of Escherichia coli, is expressed from the upstream pdh and internal lpd promoters of the pdh operon (pdhR-aceEF-lpdA). Under aerobic conditions, the specific components of the 2-oxoglutarate dehydrogenase complex encoded by the sucAB genes in the sdhCDAB-sucABCD operon are expressed from the sdh promoter. The provision of lipoamide dehydrogenase subunits for assembly into the 2-oxoglutarate dehydrogenase complex could thus be controlled by co-regulation of the lpd promoter with the sdh promoter. Here, the transcription start point of the lpd promoter was defined by primer extension analysis, and an ArcA binding site, TGTTAACAAT, overlapping the lpd promoter and matching the consensus at 8 out of 10 positions, was identified by in vitro footprint analysis. PdhR was not bound to the lpd promoter nor was ArcA bound specifically to the pdh promoter. These results support the view that co-regulation of the lpd and sdh promoters is mediated primarily by ArcA. PMID- 9868789 TI - Sphingomyelin synthesis in helminths: a minireview. AB - Sphingolipids are a diverse and ubiquitous group of lipids. They are widely distributed in parasites and a number of novel forms have been described. Sphingolipid synthesis has been investigated in the malarial parasite, cestodes, digeneans and nematodes. Although there are differences in detail, the synthetic pathways involved are similar to those found in mammals. PMID- 9868790 TI - Taxonomic problems, seasonality and histopathology of Henneguya creplini (Myxosporea) infection of the pikeperch Stizostedion lucioperca in Lake Balaton. AB - Plasmodia of a Henneguya species measuring 70-900 microns and exhibiting season dependent stages of development were detected throughout a three-year study on gill myxosporosis of Lake Balaton pikeperch (Stizostedion lucioperca (L.)). Sixty five out of 160 fish (41%) examined in the period of study were infected by the parasite. Infection was the most prevalent (48%) among pikeperch specimens exceeding 40 cm in length. The highest prevalence of infection (58%) was recorded in 1995-1996 while the lowest (30%) in 1996-1997. The youngest plasmodia appeared in April, and started to develop within the capillaries of the secondary lamellae of the gill filaments. The round or ellipsoidal plasmodia which continued their gradual growth in the subsequent months of the year achieved a size of 800-900 microns by the late autumn months, but remained in intralamellar location throughout the developmental cycle. Mature spores developed in the plasmodia by the end of winter. On the basis of their shape and size, the spores were identified as Henneguya creplini (Gurley, 1894). However, because of the uncertain taxonomy of species assigned to the genus Henneguya the taxonomic position of the parasite requires further study. The host reaction consisting of epithelial proliferation and granulation tissue formation starts around the infected secondary lamella only after the maturation of spores and the disruption of plasmodia. PMID- 9868791 TI - Helminth parasites of the rare endemic catfish, Liobagrus reini, in Japan. AB - Examinations of 29 specimens of the catfish Liobagrus reini Hilgendorf (Siluriformes: Amblycipitidae), a species endemic to Japan, collected from the Sho River, Toyama Prefecture, in central Honshu, revealed the presence of four species of helminth parasites: Plagioporus honshuensis sp. n. (Trematoda), Rhabdochona coronacauda Belouss, 1965, R. japonica Moravec, 1975 and Mexiconema liobagri sp. n. (all Nematoda). The new opecoelid trematode P. honshuensis is characterized mainly by the extent of vitelline follicles, the size ratio of the suckers (1 : 1.7-2.1), the situation of testes near the middle of the body and by the structure, size and shape of the cirrus sac. Allopodocotyle Pritchard, 1966 is considered a junior synonym of Plagioporus Stafford, 1904. The males of R. japonica are described for the first time and conspecific females are redescribed; this species is a specific parasite of L. reini. The principle hosts of R. coronacauda are cyprinids and, therefore, its finding in the catfish may indicate an accidental infection; L. reini represents a new host record. M. liobagri (only females were found) from the host's abdominal cavity is the second known species of this dracunculoid genus, being characterized by the position of the excretory pore and the number (1) of caudal processes in the female. The recorded helminths are briefly described and illustrated and problems of their systematic status are discussed. PMID- 9868792 TI - Histochemical characteristics of Gyrodactylus derjavini parasitizing the fins of rainbow trout (Oncorhynchus mykiss). AB - Histochemical studies of enzyme activities and structural elements in Gyrodactylus derjavini Mikailov, 1975 parasitizing fins of Oncorhynchus mykiss Walbaum were conducted. Marked activities of non-specific esterase, acid phosphatase, alkaline phosphatase and amino-peptidase were found in the intestinal caeca of the parasite. A strong activity of acetylcholinesterase was seen in the nervous system. Extraintestinal non-specific and eserine-sulphate resistant esterase was localized in the distal part of the hamulus sheath. Activities of peroxidase and glucuronidase were not detected. In the embryo, developing hamuli were enclosed in a sheath rich in phospholipids. Deposits of neutral lipids were sparse. The fully developed ventral and dorsal hamulus bars stained strongly for calcium. Lectin binding assays showed a mannose rich region in the cephalic duct openings, strong reactions for galactose in the glycocalyx whereas reactions for lactose were weak. These findings are discussed with respect to the parasite-host relationship. PMID- 9868793 TI - Borrelia-infection rates in tick and insect vectors accompanying human risk of acquiring Lyme borreliosis in a highly endemic region in Central Europe. AB - The methods of spatial statistics were applied to assess the geographical pattern of risk of Lyme borreliosis in Central Bohemia, the Czech Republic, based on retrospective data on disease contractions. The statistical risk was then compared at 15 selected localities with the infection challenge presented by ticks and insects carrying borreliae. Over 5,000 Ixodes ricinus (L.) ticks and 390 haematophagous dipterans were screened by direct immunofluorescence method, and the spatial and seasonal variance of infection rates were studied. Infected ticks were found at each locality throughout the warm season; in nymphs, sample infection rates ranged from 4.9% to 23.1% with a mean of 14.5% in spring, from 7.7% to 28.7% with a mean of 16.1% in summer, and from 7% to 20.6% with a mean of 13.6% in autumn. The statistical risk was found to correlate well with an average nymphal infection challenge, i.e. I. ricinus nymphal abundance x infection rate, at a given locality. Statistically significant cumulation of insect-history recalling patients into several, generally wetland, areas was ascertained; borreliae were revealed in 0.5% of the dipterans examined. PMID- 9868794 TI - Carp coccidiosis: longevity and transmission of Goussia carpelli (Apicomplexa: Coccidia) in the pond environment. PMID- 9868795 TI - In vitro growth on different cell lines of four microsporidial species infecting humans. PMID- 9868796 TI - Echinococcus multilocularis (Cestoda: Taeniidae) in the Czech Republic: the first detection of metacestodes in a naturally infected rodent. PMID- 9868797 TI - A non-excision uvr-dependent DNA repair pathway of Escherichia coli (involvement of stress proteins). AB - In UV-irradiated excision-proficient (uvr+) Escherichia coli, pre-induced by simultaneous pre-starvation for thymine (T) and amino acids (AAs), and/or a low UV pre-dose applied after prestarvation for AAs, pyrimidine dimer excision (PDE) is reduced without an adequate increase of UV sensitivity and UV mutagenesis. The unexcised lesions are tolerated by a putative repair pathway that is uvr dependent but does not involve excision. The process consists of PDE inhibition, which requires outer membrane protease OmpT, and subsequent pyrimidine dimer (PD) toleration, which may be mediated by interaction with a sister duplex using a number of SOS and stress-inducible proteins. PMID- 9868798 TI - Ultraviolet-light-evoked phase shifts in the locomotor activity rhythm of the field mouse Mus booduga. AB - The circadian rhythm of locomotor activity in the nocturnal field mouse Mus booduga has been monitored after an exposure to ultraviolet-A (UV-A; wavelength between 350 and 400 nm) radiation at various hours of the subject's cycle. The magnitude of the phase shift is used to determine the sensitivity of the circadian pacemaker underlying locomotor activity rhythm. It is observed that UV A shifts the phase of the locomotor activity rhythm in a phase-dependent manner. A phase response curve (PRC) is constructed to depict the time course and waveform of the basic oscillator's sensitivity to UV-A radiation. The UV-A PRC qualitatively resembles the PRC constructed using diffused daylight as stimulus. Furthermore, the phase response characteristics of the circadian pacemaker for UV A suggest a periodically varying sensitivity to UV-A similar to that to daylight. PMID- 9868799 TI - Photocytotoxic effect of pseudohypericin versus hypericin. AB - Pseudohypericin and hypericin, the major photosensitizing constituents of Hypericum perforatum, are believed to cause hypericism. Since hypericin has been proposed as a photosensitizer for photodynamic cancer therapy, the photocytotoxicity of its congener pseudohypericin has been investigated. The presence of foetal calf serum (FCS) or albumin extensively inhibits the photocytotoxic effect of pseudohypericin against A431 tumour cells, and is associated with a large decrease in cellular uptake of the compound. These results suggest that pseudohypericin, in contrast to hypericin, interacts strongly with constituents of FCS, lowering its interaction with cells. Since pseudohypericin is two to three times more abundant in Hypericum than hypericin and the bioavailabilities of pseudohypericin and hypericin after oral administration are similar, these results suggest that hypericin, and not pseudohypericin, is likely to be the constituent responsible for hypericism. Moreover, the dramatic decrease of photosensitizing activity of pseudohypericin in the presence of serum may restrict its applicability in clinical situations. PMID- 9868800 TI - Effect of the microenvironment on the efficiency of singlet oxygen (O2(1 delta g)) production by photosensitizing anti-inflammatory drugs. AB - The influence of the medium on the quantum yields of singlet oxygen (O2(1 delta g)) production (phi delta) by a series of photosensitizing non-steroidal anti inflammatory drugs (NSAID) derived from 2-arylpropionic acid (APA) has been investigated. Four-component oil-in-water and water-in-oil microemulsions, based on anionic and cationic surfactants, have been employed as the simplest models to mimic more complex biological environments. phi delta values have been determined by monitoring the singlet oxygen (1O2) luminescence at 1270 nm upon continuous excitation of the drugs under air-equilibrated conditions. Results indicate that phi delta values are highly affected by the medium, being higher in microheterogeneous systems than in (homogeneous) solution. Some of the anti inflammatory derivatives are very efficient 1O2 sensitizers: e.g., values of apparent phi delta as high as 0.86 (+/- 0.04) and 0.70 (+/- 0.03) have been found for tiaprofenic acid and suprofen, respectively. The location of the drugs in the interfacial region of the microemulsions combined with their high phi delta values suggest that type II reactions may play a significant role in the overall photodynamic process in more complex organized media, such as biological membranes. PMID- 9868801 TI - Phytochrome-mediated light signals are transduced to nucleoside diphosphate kinase in Pisum sativum L. cv. Alaska. AB - To clarify the molecular mechanism for the transduction of light signals in plants, we have established an in vitro system that uses crude membrane and soluble fractions of stem sections of etiolated Pisum sativum L. cv. Alaska after irradiation by red light, or sequential application of red and far-red light to the stem section. In a previous report (T. Hamada et al., J. Photochem. Photobiol. B: Biol. 33 (1996) 143-151) the labelling of proteins in membrane fraction by [gamma-32P] ATP at 0 degree C for 15 s and subsequent separation of proteins by two-dimensional electrophoresis allowed unambiguous identification of a heavily phosphorylated protein spot at 18 kDa (p18). In the present study we have confirmed the former results in the membrane fraction, and obtained the result that an increase in the phosphorylation of p18 by red-light irradiation is observed in the soluble fraction. Further, we have provided evidence that the p18 in the soluble fraction is purified and identified as nucleoside diphosphate (NDP) kinase by Western blotting, immuno-precipitation, amino acid sequencing and cDNA analysis. Purified p18 shows autophosphorylation activity and strong phosphorylating activity against myelin basic protein (MBP), a substrate of MAP (mitogen activated protein) kinase. The results show that phytochrome-mediated light signals are transduced to NDP kinase, which may elicit signals by providing high concentrations of, for example, GTP from GDT and ATP, by the autophosphorylation and by the protein kinase activity similar to MAP kinase. PMID- 9868802 TI - Photoreactions of psoralens with lecithins. AB - The formation of cyclobutane (cb) photoadducts of psoralen with a model lecithin has been shown. The adducts are formed both in ethanol solution and in micellar suspension in water. In spite of their sensitivity to various factors such as light, temperature, air, etc., they are isolated by thin-layer chromatography (TLC) and high-pressure liquid chromatography (HPLC) and characterized by mass spectrometry, NMR and UV absorption spectroscopy. The NMR analysis indicates the similarity of isomeric forms of cb adducts in lecithin to those formed with free oleic acid. PMID- 9868803 TI - Expression kinetics of the (proto) oncogenes c-myc and bcl-2 following photodynamic treatment of normal and transformed human fibroblasts with 5 aminolaevulinic acid-stimulated endogenous protoporphyrin IX. AB - In the same way as common tumour therapies can cause secondary tumour induction, photodynamic tumour therapy also shows a moderate mutagenicity. The oncogenes responsible for it can be distinguished from their proto-oncogenic precursors by an irreversible increase in their constitutive expression. Transient changes of the expression level of (proto) oncogenes can indicate the beginning of disturbances in the cell homeostasis: many of these genes have a normal function in proliferation or play a role in apoptosis. In this study, therefore, quantitative determination of the expression of the (proto) oncogenes c-myc and bcl-2 in normal and transformed human fibroblasts at different times following photodynamic treatment with 5-aminolaevulinic acid-stimulated endogenous protoporphyrin IX and low-dose irradiation has been carried out by quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The aim is to investigate if irreversibly increased (proto) oncogene expression can be found, and if expression changes are involved in cell-cycle alterations (detected in a parallel study) and in initiation of apoptotic processes. The results show: (1) no mutagenic risk, since the over-expression of c-myc and bcl-2 is transient; (2) an interaction of bcl-2 and c-myc associated with an increase of the proliferative activity of the cell cycle of transformed cells; (3) a possible role of bcl-2 in counteracting processes that could be at least precursors for apoptosis induction; and (4) higher constitutive expression of both genes in transformed than in normal fibroblasts. PMID- 9868804 TI - Enhancement of the efficiency of photodynamic therapy of tumours by t-butyl-4 hydroxyanisole. AB - The effects of photodynamic therapy (PDT) alone and in combination with 3(2)-t butyl-4-hydroxyanisole (BHA) on Ehrlich ascites carcinoma (EAC) cells have been investigated. BHA, a widely used food antioxidant, administered to the cells prior to light exposure is found to cause concentration-dependent alterations of the haematoporphyrin derivative (HpD)-based PDT. BHA (0.15 mM) causes a small (about 10%) inhibition in the rate of HpD-photosensitized injury of EAC cells. In contrast, upon increasing the concentration of BHA from 0.15 to 0.5 mM, a 1.3 fold enhancement in HpD-PDT efficiency is achieved. The cytotoxic effect on the cells treated with HpD-PDT and a higher concentration of BHA (0.5 mM) is additive. When BHA (0.5 mM) is given immediately after HpD-PDT, the combination is found to be three to four times more effective than when BHA is added to EAC cells before phototherapy. In this treatment regimen BHA acts synergistically with HpD-PDT. Such a difference in the action of BHA on the efficiency of HpD-PDT might be explained by the ability of BHA to inhibit the HpD-photosensitized destruction of some biomolecules. An enhancing action of BHA on the intensity of HpD-photosensitized death of tumour cells is also observed in vivo. Even a single dose of BHA (0.6 mM kg-1, 15 min after irradiation) causes (in an additive manner) an approximately two-fold increase in the efficiency of HpD-PDT of mice bearing Ehrlich ascites tumour (intraperitoneal transplantation). The results obtained indicate that the potentiating effect of BHA on the HpD-PDT could be caused by the impairment of the mitochondrial respiration, since there is a good correspondence between the concentration of BHA that increases the efficiency of PDT and the concentration that inhibits the oxygen consumption and dehydrogenase activity of EAC cells. The influence of BHA on the efficiency of PDT does not depend on the nature of the photosensitizer used; the effects with chlorin-e6 trimethyl ester are similar to that seen for HpD. PMID- 9868805 TI - Liposome-bound Zn (II)-phthalocyanine. Mechanisms for cellular uptake and photosensitization. AB - In the present study, cellular uptake of a liposomal formulation of ZnPc (CGP 55847) has been studied in human cervix carcinoma cells of the line NHIK 3025. The cellular uptake of ZnPc is found to be completed after 4-8 h of incubation. The maximum level of ZnPc in the cells after incubation with 1 microgram/ml ZnPc in E2a medium containing 3% serum is 60 ng/mg protein. The cellular uptake is attenuated by the presence of serum and at low temperature of the incubation medium, but the activation energy (30 kJ/mol) and fluorescence microscopic analysis of cells incubated with ZnPc at 0 degree C indicate that ZnPc is taken up into cells by a diffusion-mediated pathway. Measurements of subcellular marker enzymes have been performed immediately after light exposure of ZnPc-treated cells. The mitochondrial marker enzyme (cytochrome c oxidase) and the marker enzyme for the Golgi apparatus (UDP galactosyl transferase), but not those for lysosomes (beta-N-acetyl-D-glucosaminidase) and endoplasmic reticulum (NADPH cytochrome c reductase), are inactivated upon photodynamic treatment. These results indicate that ZnPc is mainly located in the Golgi apparatus and the mitochondria of NHIK 3025 cells. In contrast, photoactivated Photofrin is found to reduce the activity of UDP galactosyl transferase, but not that of NADPH cytochrome c reductase. The tetraphenylporphine TPPS2a and light reduce the activity of NADPH cytochrome c reductase, without influencing the activity of UDP galactosyl transferase. TPPS4 and light do not attenuate the activities of UDP galactosyl transferase and NADPH cytochrome c reductase. PMID- 9868806 TI - In vitro and in vivo porphyrin accumulation by C6 glioma cells after exposure to 5-aminolevulinic acid. AB - Several malignant tissues synthesize endogenous porphyrins after exposure to 5 aminolevulinic acid (5-ALA). The present experiments have been designed to elucidate whether the C6 glioma cell, a model cell for human malignant glioma, similarly synthesizes porphyrins when exposed to 5-ALA, and whether specific synthesis occurs when C6 cells are inoculated into rat brains to form a tumor. In this situation the blood-brain barrier may interfere with 5-ALA availability, and spreading of porphyrins with edema outside the tumor may occur. Flow cytometry is used to determine the course of cell volume and porphyrin fluorescence intensities in cultured C6 cells which are incubated in 1 mM 5-ALA. For the induction of experimental brain tumors, 10(4) untreated C6 cells are inoculated into the brains of rats. After 9 days animals receive 100 mg 5-ALA/kg body weight. Brains are removed after 3, 6, or 9 h and frozen coronal sections obtained for H/E staining or fluorescence spectography. Cultured C6 cells show a linear increase of protoporphyrin IX fluorescence after exposure to 5-ALA, which begins to plateau after 85 min. Marked fluorescence is also observed in solid and infiltrating experimental tumor. However, faint fluorescence also occurs in normal tissue, basal pia, choroid plexus, and, more obviously, in white-matter tracts bordering the tumor (maximal distance: 1.5 +/- 0.7 mm). The observations demonstrate that C6 cells synthesize protoporphyrin IX after exposure to 5-ALA in vitro and in vivo. However, when utilizing 5-ALA for fluorescence detection or photodynamic therapy of brain tumors, attention should be paid to the possibility of protoporphyrin IX occurring outside the tumor. PMID- 9868807 TI - Childbirth preparation through hypnosis: the hypnoreflexogenous protocol. AB - A verbatim protocol for the "Hypnoreflexogenous" method of preparation for childbirth is presented wherein the patient is taught to enter a hypnotic state and then prepared for labor and delivery. The method provides a "conditioned reflex" effect conducive to a positive outcome for labor and delivery by enhancing the patient's sense of readiness and control. Previous applications of the method demonstrate patients have fewer complications, higher frequency of normal and full-term deliveries, and more positive postpartum adjustment. The benefit and ultimate cost effectiveness of the method are discussed. PMID- 9868808 TI - Hypnotic technique for sedation of patients during upper gastrointestinal endoscopy. AB - A method of sedation of patients undergoing upper gastrointestinal endoscopy is described. This technique employs a variety of elements, including "pacing and leading," metaphors, use of physiological reactions to deepen the relaxation, imagery, and post hypnotic suggestions. It is a simple and effective method which does not require any preparation. It spares the need for a pharmacological sedation and obviates the possible hazards of such a sedation. The author has successfully used this technique to sedate more than 200 patients undergoing upper gastrointestinal endoscopy. The duration of examinations performed this way compares with that using conventional pharmacological sedation. However, unlike the case of pharmacological sedation, no further monitoring is needed after the completion of the examination and the patients can leave the clinic immediately to resume their activities. PMID- 9868809 TI - Using hypnosis to help deaf children help themselves: report of two cases. AB - This is a report of deaf children who demonstrated the ability to quickly learn hypnotic skills and apply them effectively to the management of their problems. The children were taught hypnosis through American Sign Language, their preferred mode of communication. As with hypnosis with hearing children, we focused upon induction with fantasy and imaginative involvement, creation in imagination of a metaphor for, or imagery of, the desired outcome, and associated sense of pride (ego-strengthening), positive expectation, and teaching self-hypnosis to emphasize the importance of repeated, daily practice. Case examples presented are an 11-year-old deaf girl who used hypnosis to eliminate multiple warts, and a 9 year-old deaf boy with mild developmental disability whose self-hypnosis skills were applied to the management of myoclonus. In the former, the clinician is also the sign language communicator and in the latter, a professional sign language interpreter and parent are both intimately involved in the communication and hypnosis process. PMID- 9868810 TI - Hypnosis enhances recall memory: a test of forced and non-forced conditions. AB - Visual memory recall in hypnosis was investigated. To address criterion shift problems in previous studies, both forced and non-forced recall procedures were used. Previous methodological weaknesses with regard to hypnotizability and hypnotic depth were also addressed. Over 300 volunteers were screened for hypnotizability using the Harvard Group Scale of Hypnotic Susceptibility: Form A (Shor & Orne, 1962). Final high and low hypnotizability groups were selected using the Stanford Hypnotic Susceptibility Scale: Form C (Weitzenhoffer & Hilgard, 1962). Participants in each hypnotizability group were randomly assigned to either forced or non-forced recall conditions and to hypnosis or waking conditions. Participants were shown 60 slides of line drawings and then tested immediately in 3 recall periods. Analysis of variance results showed that those exposed to hypnosis and to a forced recall procedure were significantly more confident of their responses to correct items than those exposed to a non-forced recall procedure or a waking condition. Participants exposed to hypnosis and forced recall procedures recalled more correct items than those exposed to a waking condition. The findings support the hypermnesic effects of hypnosis when participants are required to provide a fixed number of responses. PMID- 9868811 TI - Metaphor dialog in psychotherapy. AB - I suggest a psychotherapeutic approach which involves extended allegorical conversations between therapist and patient, and present 2 case examples. The approach, Metaphor Dialog (MD), can be described as a 4-way conversation among the conscious and unconscious aspects of both persons' psyches. Although it typically begins with patient-generated material, MD often involves empathic, intuitive, and spontaneous interventions by the therapist which may depart significantly from the metaphoric material overtly presented by the patient. I argue that such an approach represents a beneficial balance between existing metaphor-based techniques which emphasize either patient-generated or therapist generated content. MD may offer important increases in therapeutic potency in contrast to techniques in which attention to the metaphors developed by 1 or the other party are assumed to be more effective or appropriate. PMID- 9868812 TI - The need for clinically relevant research. PMID- 9868813 TI - Training in clinical hypnosis, a historical perspective: the Montana experience. AB - When Dr. D. Corydon Hammond invited me to write a paper for this symposium on hypnosis training, it was obvious that its greatest contribution could be a description of the rather intensive program developed over 2 decades at the University of Montana. However, it became apparent that the Montana program was only part of some 50 years experience developing professional education in hypnosis, psychotherapy, and graduate training in general. At the risk of being accused of immodesty I must therefore write much of this contribution in the first person. PMID- 9868814 TI - Clinical training in Ericksonian hypnosis. AB - The authors place great value on the role of experiential learning in their clinical trainings on Ericksonian approaches to hypnosis. This article describes several of the principal skill-building exercises used in their training programs. These highly practical exercises are intended to facilitate: (a) developing sensory acuity, (b) developing flexibility in forming and delivering suggestions, (c) employing naturalistic methods, (d) employing reframings, and (e) embedding suggestions. A rationale and goal for each exercise are also presented in order to help create an appropriate context for its use. PMID- 9868815 TI - Professional training in the practice of hypnosis--the Australian experience. AB - Increasingly around the world as clinicians and researchers alike become more sophisticated in their understanding of the hypnotic process, there is developing a conviction that the hypnotic state or process itself poses no inherent dangers for patients but that its inexpert use may. The solution to prevent potential patient harm is to ensure that all clinicians of whatever discipline have adequate and appropriate clinical training prior to being allowed to practice. Since 1985, in all the Australian states, the appropriate disciplines, already licensed to practice their profession, have completed 2 years part-time academic and clinical training in the nature and nuances of hypnosis (30 hours), practical experience of direct and indirect approaches, and supervised clinical case management (50 hours). At the end of the training they complete a 3 part examination of competence (clinical cases reports, written examination, and oral examination) in order to be accepted as members of the Australian Society of Hypnosis. Only by completing this training and peer reviewed assessment are clinicians able to receive the backing of the Society and recommendation to patients of their competence as clinicians using hypnosis. Recently in the states of Victoria and South Australia (and soon in the state of Queensland), the training program of the Australian Society of Hypnosis has been incorporated into a university diploma course, giving formal academic recognition to the approach to training. The University of Melbourne diploma course will be discussed with a view to illustrating these recent developments. PMID- 9868816 TI - Future developments in professional hypnosis training: a review and critique of available training with consideration for development of a nationally coordinated program. AB - This article seeks to review current training models of hypnosis within a framework of basic distinctions between 2 primary goals of professional training programs and 3 primary aspects of professional training models. We focus on a description and critique of available training programs in the context of progress which has been achieved in the teaching of professional hypnosis. Programs within and the United States, Australia, and the Netherlands are reviewed, along with efforts to implement an "Americanized" version of training which incorporates desirable aspects of these latter programs. Suggestions are also made for what might be gained from adoption of a nationally coordinated program of training in professional hypnosis. PMID- 9868817 TI - Standards of training in clinical hypnosis: preparing professionals for the 21st century. AB - During the past 40 years, there has been increased interest in clinical hypnosis among qualified health care professionals. However, training programs in the past have been quite variable. This has led to the identification of a need for an accepted standard curriculum in clinical hypnosis. The present research surveyed professionals regarding qualifications and training in clinical hypnosis. The continued growth of clinical hypnosis among professionals may ultimately rest upon the development of an educational standard that integrates clinical skills and knowledge from research. PMID- 9868818 TI - A graduate school curriculum in clinical hypnosis. AB - The literature suggests a growing interest in hypnosis training. With this growing interest, there has been an increase in training opportunities for the interested clinician. However, much of that training is in the form of a single course or workshop, or a series of single workshops. Many programs only offer hypnosis courses when the appropriate faculty is interested in offering such a course. Other programs offer a minimal exposure to hypnosis, less than a minimal degree of training as suggested by ASCH. This paper describes a formal, integrated curriculum in clinical hypnosis that is offered through a psychology graduate school. An integrated program offers students and practicing professionals the opportunity to learn hypnosis in an environment that offers ongoing supervision and support of their long term development. PMID- 9868819 TI - Facilitating facilitators' facilitation: experience with a model for teaching leaders of hypnosis practice groups. AB - We developed a 6 hour training seminar for leaders of small group practice sessions integral to introductory hypnosis workshops. It has been offered annually for 3 years to prepare new faculty for our Introductory Workshop. Applicants met specific experience criteria. Participation was limited to 8 to match and model 8 learners/small group in workshops. Learners reviewed principles of adult education, group dynamics, and guidelines for leaders. Each had the opportunity to role-play small group participant and small group leader in two 3 hour evening periods a week apart. Twenty-four learners participated. Some had taught before. Most were new to teaching small groups and desired specific training. Extensive evaluation of the training was integral to the process. Training was evaluated by examining ratings of leaders by participants in our Annual Introductory Workshop. Ratings used the same parameters taught and evaluated during the training seminar. The training curriculum is presented with the results of evaluations of group leaders' performances in the Annual Workshop. Ratings of "experienced" leaders who did not participate in the training are compared to those of newly trained teachers. Leaders with training plus experience were more effective then leaders with many years of experience. PMID- 9868820 TI - The nurse-patient covenant and the imperative to care. PMID- 9868821 TI - A tale of two centuries: voices of the past and present. AB - This article compares the reported experiences of children and adolescents receiving inpatient treatment in contemporary psychiatric hospital settings with those of women institutionalized in mental hospitals from 1840 to 1945. Qualitative data include case study material from former psychiatrically hospitalized patients and archival narratives. Analysis includes matching themes from in-depth interviews with archival narratives. Nine representative experiences emerged and are presented in a typology under broader themes of structure, process, and outcome. Similarity of patient experiences over two centuries leads to questions about the meaning of advances in understanding mental illnesses in the context of attitudes, biases, and staff behavior toward patients that remains disturbingly unchanged. PMID- 9868822 TI - The psychiatric clinical nurse specialist/nurse practitioner: an example of a combined role. AB - The purpose of this article is to provide an historical overview of the clinical nurse specialist and nurse practitioner roles in psychiatric nursing. This article is written as a follow-up to that of Dyer, Hammill, Regan-Kubinski, Yurick, & Kobert, (1997), Archives of Psychiatric Nursing, 11, 2-11, in which a new paradigm for delivering comprehensive mental and general health services is conceptualized, i.e., the psychiatric-primary care nurse practitioner role. In this article an alternative model is presented: the psychiatric clinical nurse specialist/nurse practitioner role. Similarities and differences in the two roles are discussed. PMID- 9868823 TI - Psychiatric nursing education: doing the impossible? AB - Historically, psychiatric nursing has struggled to attract sufficient numbers of graduates of comprehensive nursing programs. Limited available research suggests that psychiatric-nursing education can produce more positive attitudes towards psychiatric-nursing practice and the care of the mentally ill. This article describes the results of a research project undertaken in Victoria, Australia, to extend existing knowledge by determining the impact of education in augmenting interest in psychiatric nursing as a future career option. The results of this quasi-experimental study suggest education can significantly increase the popularity of psychiatric nursing as a profession. PMID- 9868824 TI - Depression and codependency in women. AB - Seven million American women are depressed, and 40 million Americans, primarily women, have been labeled as codependent. This study aimed to identify the prevalence of codependency in women undergoing treatment for depression, examine the relationship between codependency and depression, and determine which of the symptoms of codependency are most highly predictive of depression scores. Depression and codependency were measured in a sample of 105 depressed women by using the Beck Depression Inventory and the Codependency Assessment Tool. Descriptive statistics, Pearson's Product Moment Correlation, and multiple regression were used for analysis. Of these depressed women, 36% were moderately to severely codependent. Depression and codependency were strongly related, with the significant gamma = .92 (P < .001). Of the codependency subscales, Low Self Worth and Hiding Self correlate most strongly with depression; Other Focus/Self Neglect added the least-independent--explanatory power. Thus, future research should be directed toward the relationship of codependency to power, alienation of self, and personality disorders. PMID- 9868825 TI - The relationship between self-induced water intoxication and severity of psychiatric symptoms. AB - The purpose of this study was to test a theoretical framework that proposed a relationship between severe psychiatric symptoms and self-induced water intoxication (SIWI) by using reliable and valid measures. Twenty of 28 community dwelling individuals with severe mental illness (SMI) in the sample exhibited excess fluid consumption as reflected by a mean urine specific gravity of 1.003 mEq/L. The psychometric measures included the Positive and Negative Syndrome Scale (PANSS), the Self Deficit Syndrome Scale (SDSS), and the Spielberger State trait Anxiety Scale (STAI From X-1). A significant relationship was found at p .01 between severity of psychotic symptoms and severity of SIWI and between severity of SIWI and psychopathology symptoms within the PANSS. Anxiety was higher in those with SIWI before fluid loading compared with those with less excess fluid intake (i.e., USG < 1.010), and anxiety decreased from morning to evening in those with SIWI compared with those who did not exhibit excess fluid intake. The findings revealed a strong relationship between SIWI and severe psychiatric symptoms, including psychosis and a broad range of psychiatric symptoms. The findings provided initial support for the proposed theory, and consideration needs to be given to the development of interventions to augment existing treatment of fluid control. PMID- 9868826 TI - Promoting enjoyment and self-belief through work rehabilitation. AB - There has been a resurgence of interest in work rehabilitation in recent years. A variety of service models are in operation in a number of countries, all, to varying degrees, assisting mental-health clients achieve an optimum level of employment participation. However, little research has been conducted to examine how (and if) such schemes are structured to promote psycosocial rehabilitation. To explore such issues, the author carried out a pilot case study of a work rehabilitation organization in Lincolnshire, United Kingdom. Two of the main themes, that emerged during the study, Hedonic Experiences and Self-Efficacy, are discussed in this article. PMID- 9868827 TI - Kinds of therapy for persons with psychiatric disorders and their families. PMID- 9868828 TI - [Total and selective deficiency of myeloperoxidase in peripheral monocytes lead to diagnosis of acute myelomonoblastic leukemia]. PMID- 9868829 TI - [Prostatic cancer in the Grand Duchy of Luxembourg. Role of PSA]. AB - We reviewed the trends in prostate cancer incidence and mortality in Luxembourg between 1983 and 1995 to discuss the importance of total and free PSA in early detection. The study was performed on all the new cases recorded by the National Cancer Registry (Registre Morphologique des Tumeurs). Total and free PSA were measured with the automated Immulite System (DPC, Los Angeles) using a chemoluminescent immunometric assay. The performance of free-to-total serum PSA was analysed by a hospital based study of 113 patients (55 PC, 58 BPH). The age standardized incidence rate increased from 29.3/100,000 in 1983 to 71.5/100,000 in 1995. Mortality rates only changed slightly. The widespread use of PSA testing from 1988 on is probably the main cause of this incidental increase; however no major changes in the age-specific-incidence have been found suggesting the absence of a systematic screening policy by the PSA. The superiority of free-to total serum PSA ratio in discriminating between cancer and benign condition was confirmed. Early health-conscious man over 50 should be proposed prostate cancer screening by digital rectal examination and PSA. However a systematic screening policy cannot been recommended since a benefit in survival after early treatment has not yet been proven. PMID- 9868830 TI - [Spongiform encephalopathies]. AB - Mad cow crisis (Bovine Spongiform Encephalopathy) had many consequences on the regulatory framework in several fields. It is now obvious that the agent of bovine spongiform encephalopathy is transmissible to man, provoking a new condition which differs from the sporadic from of Creutzfedt-Jakob Disease (CJD) called variant or new variant (vCJD). This paper tries to review the most important new findings in the scientific field of spongiform encephalopathies. PMID- 9868831 TI - [Spontaneous regression of pulmonary metastases in renal cancer]. AB - Spontaneous regression of lung-metastases in kidney cancer (a case report) A case is reported of a 56-year old woman with renal cell carcinoma operated and treated adjuvantly for a year with Interferon alpha. Ten months thereafter the patient had histologically proven a large pulmonary metastatic disease that regressed spontaneously. Duration of the spontaneous remission was 8 months. PMID- 9868832 TI - [Results of field studies using Stellamune mycoplasma vaccine in Luxembourg]. AB - Enzootic pneumonia caused by Mycoplasma hyopneumoniae is responsible for major economic losses in pig herds worldwide. M. hyopneumoniae can also act as a primary agent of swine respiratory disease followed by bacterial or viral infections. Vaccines have been developed to prevent enzootic pneumoniae. In this experiment, an inactivated vaccine against mycoplasma, also sold under the name Stellamune mycoplasma (Pfizer Animal Health) was used in 2 farms with a known history of M. hyopneumoniae infection. The objective of the study was to evaluate the effect of the product on lung lesions caused by M. hyopneumoniae and on economic performance of the pigs under field conditions. PMID- 9868833 TI - [The threat of premature labor: new aspects for management]. AB - Genital inflammation may play a major role in the pathogenesis of preterm labor. Screening and early treatment of subclinical genital tract infections (bacterial vaginosis, heavy group B streptococci colonization, primary genital HSV infection and other silent intra-uterine infections) seem to offer promise for the prevention of preterm labor. New factors have been studied in order to appreciate their benefit in the evaluation of the risk of preterm labor. None of these biologic markers (fetal-fibronectin, maternal interleukin-6, vaginal pH measuring) have enough sensitivity to permit efficient screening. Home uterine activity monitoring seems to be interesting for early identification of women with increased risk of preterm delivery, but can't be used on a large scale because of its costs. New tocolytic agents are investigated in order to protect from an adverse outcome. Atosiban exhibits more oxytocin selective and antagonistic activity without side-effects, and nimesulide seems to have a lack of effects on fetal functions. PMID- 9868834 TI - Sports and children: consensus statement on organized sports for children. FIMS/WHO ad Hoc Committee on Sports and Children. PMID- 9868835 TI - Tetanus toxoid immunization coverage among women in zone 3 of Dhaka city: the challenge of reaching all women of reproductive age in urban Bangladesh. AB - Neonatal tetanus is still an important public health problem in both urban and rural Bangladesh, with an estimated 41,000 cases occurring annually. This article analyses the coverage of tetanus toxoid (TT) immunizations among women of reproductive age in Zone 3 of Dhaka City in 1995. Although 85% of women with a child under 1 year of age had received two TT immunizations, only 11% of women of reproductive age had obtained the complete series of five TT immunizations and only 52% of women of reproductive age had received one or more TT immunizations. Access to TT immunization, as defined by having had at least one such immunization, was lower among women aged over 30 years and also among those aged under 20 years, especially those who were not yet married or who had not yet become pregnant. Characteristics associated with TT immunization status included the following: educational level of the woman, distance from the nearest immunization centre, and level of contact with family planning field workers. Additional characteristics that influenced women's TT immunization status included age, marital and working status, recency of migration from rural to urban area, and number of children. The relationships were complex and varied depending on the number of TT immunizations received (one or two) and on the type of analysis being carried out (bivariate or multivariate). The findings point to the need for a broad-based campaign to promote access to TT immunization as well as to promote the completion of all five TT doses in Bangladesh. Reducing missed opportunities for promotion of immunization as well as targeting home visitation of women in need of additional immunizations constitute further approaches to improving coverage. Although TT coverage rates were only marginally lower among women in slum households, such women were more likely than those in non-slum households to be pregnant and hence more likely to bear a baby at risk of neonatal tetanus. Furthermore, the environment of slum households, where deliveries normally take place, is more conducive to the development of neonatal tetanus among unprotected neonates; a strategy of focusing on slum households is therefore also needed. PMID- 9868836 TI - Estimating costs of programme services and products using information provided in standard financial statements. AB - The financial viability of programme services and product offerings requires that revenue exceeds expenses. Revenue includes payments for services and products as well as donor cash and in-kind contributions. Expenses reflect consumption of purchased or contributed time and materials and utilization (depreciation) of physical plant facilities and equipment. Standard financial reports contain this revenue and expense information, complemented when necessary by valuation and accounting of in-kind contributions. Since financial statements are prepared using consistent and accepted accounting practices, year-to-year and organization to-organization comparisons can be made. The use of such financial information is illustrated in this article by determining the unit cost of cataract surgery in two hospitals in Nepal. The proportion of unit cost attributed to personnel, medical supplies, administrative materials, and depreciation varied significantly by institution. These variations are accounted for by examining differences in operational structure and capacity utilization. PMID- 9868837 TI - Use of sentinel lambs to survey the effect of an education programme on control of transmission of Echinococcus granulosus in South Powys, Wales. AB - In this article the effects of an education programme (area II) on transmission of Echinococcus granulosus from dogs to sheep in Wales and of substitution of the education programme by a 6-weekly anthelmintic control programme (area I) are compared with the situation in an area where no control interventions had occurred (area III). The education programme failed to prevent transmission of E. granulosus to sentinel lambs examined at 15 months of age, 6%, 4%, and 10% of which were infected in areas I, II, and III, respectively. Educational efforts did, however, show some positive effects; for example, significantly more farmers (87-98%) in areas I, II, and III used praziquantel to treat their dogs compared with 39% of farmers in a lowland area in the east of England where E. granulosus is absent. In particular, the interval between treatments of dogs was significantly shorter in areas targeted with education programmes, and 38% of farmers in area I treated dogs at a 4-6 week interval, as did 17% in area II and 10% in area III, compared with only 3% in the lowland area. Also, more dogs in Wales (65-88%) were treated at an interval of < or = 3 months, whereas most of the dogs in the lowland area (64%) were treated at intervals of > or = 6 months. The shorter treatment intervals with praziquantel may account for the significantly fewer positive coproantigen tests among dogs in area I (6.3%) and area II (5.6%) compared with area III (23.9%). PMID- 9868838 TI - Cost-effectiveness analysis of tuberculosis control policies in Ivanovo Oblast, Russian Federation. Ivanovo Tuberculosis Project Study Group. AB - Many of the current tuberculosis control programmes in the Russian Federation are based on costly strategies which are underfunded and use long, individualized treatment regimens. This article compares, using a cost-effectiveness analysis, the new WHO strategy implemented in the Ivanovo Oblast (case-finding among symptomatic patients (SCF) and shorter regimens) and the old strategy (active screening of the asymptomatic population (ACF) and longer regimens). The cost per case cured was calculated at different levels of cure rate (45-95%) using three scenarios to describe the new WHO strategy (use of WHO-recommended regimens and three options at increasing rates of admission) and a fourth scenario to describe the old strategy (all patients admitted for the whole treatment and longer regimens). The cost per case detected was determined by calculating the following: yield of the new and old strategy (number of examinations necessary to diagnose one case); cost of the diagnostic process; multiplying yield per cost according to the three scenarios describing the new WHO strategy and a fourth scenario describing the old strategy. In the Ivanovo Oblast the cost per case cured, at 85% cure rate level, ranged from US$ 1197 (new strategy, scenario 1 without food) to US$ 6293 (old strategy, scenario 4) the cost per case detected ranged from US$ 1581 (new strategy, scenario 1) to US$ 4000 (old strategy, scenario 4). Significant savings can result from shifting towards the new WHO strategy. Decision-makers and health administrators should be responsible for re investing the financial and human resources mobilized by the adoption of cost effective strategies within the TB control programme. PMID- 9868839 TI - Knowledge, attitudes, and practices about kala-azar and its sandfly vector in rural communities of Nepal. AB - Reported are the results of a study of the knowledge, attitudes and practices (KAP) about kala-azar of the inhabitants of two villages (Titaria and Haraincha) situated in terai (plain) areas of Nepal. The villagers had poor knowledge about the transmission of kala-azar, with most villagers perceiving that mosquitos, instead of sandflies, were responsible for transmission of the infection. Most also failed to recognize the common symptoms of kala-azar. The majority of the respondents, 78.9% in Titaria and 48.4% in Haraincha, were aware that the condition can be treated, while fewer than 2% believed that it cannot be treated at all. More than 58% of villagers in Titaria and 36.8% in Haraincha used bednets. The residents of both villages were highly responsive to a programme to spray houses with insecticides. Fewer than 5% of respondents slept outdoors in farm outhouses and these individuals did not take any personal vector control measures. The results of this study show the importance of understanding the beliefs and practices of communities in the successful planning and implementation of kala-azar control activities in Nepal. PMID- 9868840 TI - Suppressant effect of human or equine rabies immunoglobulins on the immunogenicity of post-exposure rabies vaccination under the 2-1-1 regimen: a field trial in Indonesia. MAS054 Clinical Investigator Group. AB - WHO's reference protocol for post-exposure rabies vaccination advises five intramuscular injections on days 0, 3, 7, 14, and 30; in addition, rabies immunoglobulins (RIG) must be given to serious cases of exposure (grade III severity). Some studies indicate that these immunoglobulins suppress the immunogenicity of rabies vaccine when administered according to an alternative protocol of four injections (2-1-1) on days 0, 7, and 21, which was therefore not recommended for grade III exposures. To test this effect, we conducted a multicentre study in Indonesia using three groups of subjects. One group received only the Vero-cell rabies vaccine (PVRV, Verorab, usual commercial lot) according to the 2-1-1 schedule. The second and third groups received the same schedule of PVRV, plus either equine rabies immunoglobulins (ERIG, 40 IU/kg body weight) or human rabies immunoglobulins (HRIG, 20 IU/kg body weight). Our results confirmed the immunoglobulin suppressant effect, which was more pronounced with human than equine immunoglobulins. In both groups receiving immunoglobulins, the seroconversion rates did not reach 100% on day 28 and the geometric mean antibody titre was lower. Thus, WHO's recommendation in 1992 of the reference protocol plus immunoglobulins for severe cases is substantiated by these results in Indonesian subjects. If the 2-1-1 regimen is chosen by the treating physician and immunoglobulins are indicated, preference should be given to purified equine RIG, which also costs less than human RIG. PMID- 9868841 TI - Eradication of schistosomiasis in Guangxi, China. Part 2: Political economy, management strategy and costs, 1953-92. AB - Reported are the results of a study of the political economy, management, and costs of the successful Guangxi schistosomiasis eradication programme, spanning 40 years from 1953 to 1992. For this purpose we analysed all government data and memoranda on the policy, management, technical support, finance, and the control strategy of the programme. We also interviewed many local staff involved in the programme over the 40-year period and obtained cost data from annual county-level records on seven major categories of variable costs. Schistosomiasis control in Guangxi began with one of the first examples of community participation and rapid assessment in public health history--the use of pre-franked envelopes to return disease questionnaires and suspect snails from rural areas. This approach quickly and accurately delineated the endemic area. This was Mao Zedong's "mass line", incorporating ideas and knowledge from peasants directly into services run for and by them, here the schistosomiasis control programme. Recognition by China's leaders that schistosomiasis was a great economic burden, steadfast prioritizing of the programme over 40 years, local innovative scientific study, agricultural and environmental focus on eradicating the snail hosts and boosting rural production, and mass community education and support were all key factors in the final success. Local leaders motivated programme staff and everyone involved knew the objectives. The programme was always multisectoral, with policy developed centrally, and strategy and collaboration encouraged and rewarded at the grass roots. These features explain how a very poor autonomous region such as Guangxi finally eradicated schistosomiasis, spending less than US$ 0.50 per protected citizen per year; it is remarkable that the disease and snails were initially found across a large area of complex environments and modern drugs such as praziquantel were not available for most of the 40-year period. The lessons from Guangxi can be adapted elsewhere and should encourage other areas to control endemic schistosomiasis using methods devised to suit the local culture and geography. PMID- 9868842 TI - HIV epidemic in Punjab, India: time trends over a decade. AB - Although Bombay (Mumbai) appears to be the main focus for acquired immunodeficiency syndrome (AIDS) in India, rapid spread has occurred through other major cities as well. The first AIDS patient in the northern state of Punjab was reported in May 1987. The present study, spanning a decade, shows that the incidence in high-risk groups increased from 3 per 1000 in 1987 to 59 per 1000 in 1997, 73% of the cases being in the third and fourth decades of life, i.e. the most productive years. The male to female ratio was 3.1:1, and 29% of the patients had the full-blown disease. A total of 80.5% acquired the infection heterosexually and only 2% of the patients were intravenous drug users. The percentage of patients acquiring infection through blood, blood products, and haemodialysis dropped from 20% in 1987 to 5% in 1997, but the cumulative figure was still 12%. The intervention programme launched by the National AIDS Control Organization (NACO) appears to have had little impact on the epidemic. There is an urgent need therefore for more interactive programmes that include education concerning the modes of spread, course, financial implications and fatal outcome of the disease, instead of passive dissemination of information by posters and the media. PMID- 9868843 TI - Snake-bites: appraisal of the global situation. AB - The true global incidence of envenomations and their severity remain largely misunderstood, except for a few countries where these accidents are rare or are correctly reported. Nevertheless, this information is essential for drawing up guidelines for dealing with snake-bites, to plan drug supplies, particularly antivenin, and to train medical staff on snake-bite treatments. Since the comprehensive review by Swaroop & Grab in 1954 no global survey has been carried out on snake-bite epidemiology. The present article is an attempt to draw the attention of health authorities to snake envenomations and urges them to prepare therapeutic protocols adapted to their needs. PMID- 9868845 TI - Statement on oral cholera vaccination. PMID- 9868846 TI - Neighborhood deterioration, religious coping, and changes in health during late life. AB - Research indicates that older adults who reside in deteriorated neighborhoods experience more physical health problems than elderly people who dwell in more favorable living environments. The purpose of this study is to see whether the deleterious effects of run-down neighborhoods are reduced for older people who use religious coping responses. Data from a nationwide longitudinal survey of elderly people suggest that the noxious impact of living in a dilapidated neighborhood on changes in self-rated health over time is offset completely for older adults who rely heavily on religious coping strategies. In contrast, significant stress-buffering effects failed to emerge when functional disability served as the outcome measure. PMID- 9868847 TI - Facility-level outcome performance measures for nursing homes. AB - Risk-adjusted nursing home performance scores were developed for four health outcomes and five quality indicators from resident-level longitudinal case-mix reimbursement data for Medicaid residents of more than 500 nursing homes in Massachusetts. Facility performance was measured by comparing actual resident outcomes with expected outcomes derived from quarterly predictions of resident level econometric models over a 3-year period (1991-1994). Performance measures were tightly distributed among facilities in the state. The intercorrelations among the nine outcome performance measures were relatively low and not uniformly positive. Performance measures were not highly associated with various structural facility attributes. For most outcomes, longitudinal analyses revealed only modest correlations between a facility's performance score from one time period to the next. Relatively few facilities exhibited consistent superior or inferior performance over time. The findings have implications toward the practical use of facility outcome performance measures for quality assurance and reimbursement purposes in the near future. PMID- 9868844 TI - Epidemiology of rotavirus diarrhoea in Africa: a review to assess the need for rotavirus immunization. AB - Rapid progress towards the development of rotavirus vaccines has prompted a reassessment of the disease burden of rotavirus diarrhoea in developing countries and the possible impact of these vaccines in reducing diarrhoeal morbidity and mortality among infants and young children. We examined the epidemiology and disease burden of rotavirus diarrhoea among hospitalized and clinic patients in African countries through a review of 43 published studies of the etiology of diarrhoea. The studies were carried out from 1975 through 1992, and only those in which a sample of more than 100 patients with diarrhoea were specifically screened for rotavirus by using an established diagnostic test were included. Rotavirus was detected in a median of 24% of children hospitalized for diarrhoea and in 23% who were treated as outpatients; 38% of the hospitalized patients with rotavirus were < 6 months and 81% were < 1 year of age. Rotavirus was detected year-round in nearly every country and generally exhibited distinct seasonal peaks during the dry months. In 5 countries where rotavirus strains had been G typed, 74% of strains were of one of the four common serotypes (G1 to G4), G1 was the predominant serotype, and 26% were non-typeable. This cumulative experience from 15 African countries suggests that rotavirus is the most important cause of severe diarrhoea in African children and that most strains in circulation today belong to common G types that are included in reassortant vaccines. Wherever large numbers of cases of rotavirus diarrhoea occur early in infancy, immunization at birth may protect the children before their first symptomatic infection. PMID- 9868849 TI - The influence of risk adjustment methods on conclusions about quality of care in nursing homes based on outcome measures. AB - This study compares quality rankings of about 550 nursing homes in New York State, based on several outcome measures and differing risk adjustment methods. The outcomes were decline in functional status, worsening decubiti, and prevalence of physical restraints. Measures were constructed from PRI data, which are similar to MDS data. We found substantial disagreement on quality ranking across measures due to differences in the scope of risk adjustment. Insufficient risk adjustment of outcome measures may, therefore, lead to inappropriate classification of nursing homes as either poor-quality or high-quality homes. This has implications for state quality oversight, providers' reputations and patients' choice. PMID- 9868848 TI - Retiree health benefits: availability from employers and participation by employees. AB - Data from the September 1994 Current Population Survey are used to examine the factors associated with early retirees' access to offers of retiree health insurance (RHI) and their likelihood of accepting these offers. I find large differences in offer rates, with those with pensions and from large firms much more likely to receive an offer, and women and non-Whites less likely. Even among those with an offer of RHI, more than a quarter do not accept the offer. Early retirees who have lower incomes and lack pensions are significantly less likely to accept an offer of RHI, suggesting that policies that increase offers are not sufficient to increase coverage for early retirees. Barriers to accepting offers need to be considered. PMID- 9868851 TI - Intensive care, old age, and the problem of death in America. AB - This article, part of a larger anthropological investigation of how death occurs in the hospital, explores the relationship of elderly deaths in the intensive care unit to the cultural conversation about the desire for "death with dignity." Based on participant observation, it provides three case studies that focus on the unfolding of events surrounding patient treatment, decision making, and family involvement. The cases are interpreted in the context of four sources of the culturally defined "problem" of death: (a) how medicine operates as the dominant conceptual framework for understanding both old age and death; (b) the power of the technological imperative to determine events; (c) ambivalence regarding end-of-life goals; and (d) the incommensurability of lay and medical knowledge. PMID- 9868850 TI - Hourly care received by people with Alzheimer's disease: results from an urban, community survey. AB - People with Alzheimer's disease living in a defined community in New York City were identified and assessed on two occasions to determine the number of hours of activities of daily living (ADL) care they received. Nearly half received all care hours from informal sources; however, a quarter received all ADL care hours from formal sources. The mean number of daily ADL hours in the sample was 9.7, of which 4.2 came from formal sources. This reflects the extensive use of Medicaid based home care services in New York City. As dementia worsened, substitution of formal for informal care was rare, but formal care assumed a greater proportion of total care hours. PMID- 9868852 TI - A conceptual model for the mobility patterns of nursing home admissions. AB - This article draws from the literature in long-term care, demography, and geography to develop an initial conceptual framework to explain variations in the patterns of nursing home moves. The conceptual framework is bolstered by preliminary data from state censuses of nursing homes that asked about the origins of current nursing home residents. The data suggest that a relatively high proportion of nursing home residents move to a different county in the process of admission to a nursing home. The research and applied implications of the framework are discussed. PMID- 9868853 TI - A standardized menu for Goal Attainment Scaling in the care of frail elders. AB - Goal Attainment Scaling (GAS), an individualized measurement approach, is particularly attractive for the evaluation and care planning of frail elders, who often experience multiple, complex problems. Some service providers find GAS too unwieldy for routine use. A modified version of GAS that uses a standardized menu of goals and attainment levels has been developed by the Geriatric Assessment and Rehabilitation Unit of a regional referral hospital in Nova Scotia, Canada. This article reviews the development, implementation, benefits, and challenges of the standardized menu, as well as the results of a retrospective descriptive study of its measurement properties. The standardized menu appears to be a feasible, valid, and responsive alternative to traditional GAS, while retaining much of its individualized nature. PMID- 9868854 TI - Participatory design of an Internet-based information system for aging services professionals. AB - The Michigan Aging Services System, a World Wide Web-based information system, was developed for practicing gerontologists in the state of Michigan. A participatory design framework adapted from the software development literature guided the development process. Users from Michigan's aging network participated in the development process. Design data were collected using multiple techniques. The system's content was developed with extensive input from actual users. The content emerged into four main categories: News and Current Events, General Information, Policy and Program Information, and Population Information. The system content is briefly compared to that of World Wide Web sites published by other state units on aging. PMID- 9868855 TI - Is the world clinical congress still relevant in the new millennium? PMID- 9868856 TI - Papillary carcinoma in a thyroglossal duct remnant--a review of thyroglossal surgery in a regional centre in the west of Ireland. AB - A review of thyroglossal duct remnants presenting to a regional centre in the West of Ireland was undertaken. Over a 15 yr period, 25 patients were operated on for duct remnants. The mean age was 19.6 yr and ranged from 3 to 68 yr. There were 16 (64 per cent) males and 9 (36 per cent) females. Seventeen (68 per cent) patients were less than 20 years at the time of surgery. Four patients presented with a sinus and the remainder with a cystic lesion. Two patients experienced recurrent disease. One patient, a 41 yr old female, had a papillary carcinoma of a thyroglossal cyst. There were no clinical features distinguishing this patient from those with benign cystic remnants of the thyroglossal duct. The possibility of carcinoma in older patients, in particular females, presenting with thyroglossal cysts emphasises the importance of performing a formal Sistrunk's operation. It reduces the risk of recurrence of the cyst and may reduce the risk of recurrence of the tumour as the duct may provide a route for the spread of tumour. PMID- 9868857 TI - Bleeding times and the antithrombotic effects of high-dose aspirin, hirudin and heparins in the rat. AB - Bleeding can occur unexpectedly during antithrombotic therapy. Impaired haemostasis is commonly measured by the bleeding time. We measured it by 3 methods in controls and in anticoagulated animals and related it to their antithrombotic status. In 42 control rats template, tail-tip transection and needle occlusion bleeding times correlated poorly (r = 0.05-0.34). The template method had the best range (mean 126.97 +/- SEM secs) and consistency. In 10 control animals it correlated mildly (r = 0.55) with venous thrombus in the same animal. Thrombus was measured by its weight deposited on platinum wires (2 cm long, 0.4 mm diameter) set in vein and in artery for 1 h. In respective groups of 10 rats, a decrease of mean thrombogenesis was obtained using aspirin, heparin and low molecular weight heparin in 2 dosages and hirudin in 1 dosage. The drugs reduced mean venous thrombus by 13-86 per cent of the mean control thrombus, and prolonged the mean template bleeding time by 29-199 per cent. The ranking of the drugs according to their increase of template bleeding time was virtually the same as the ranking given by their reduction of thrombus weight (Spearman rank coefficient 0.81, sig 0.007). The transection test produced a similar ranking and similar correlation with thrombus (0.71, sig 0.049). Low molecular weight heparin induced the greatest thrombus reduction (39 per cent) for least prolongation of bleeding time (24 per cent). Arterial thrombus was more variable. The bleeding times and thrombus weight were measured in each animal of 2 groups given aspirin, the template method correlating mildly with venous thrombus reduction (r = 0.23, 0.58 respectively), the transection method with arterial (0.74, 0.45) and the occlusion test poorly with either (0.13, 0.22). Bleeding time lengthens with increasing antithrombotic effect of drugs, but not in direct proportion, nor similarly with each drug. PMID- 9868858 TI - Long term results of polytetrafluoroethylene in above knee femoropopliteal bypass for critical ischaemia. AB - Ninety-six consecutive above-knee femoropopliteal bypasses, using polytetrafluoroethylene (PTFE) preferentially, were performed for limb threatening ischaemia. Cumulative primary graft patency was 68 per cent, 49 per cent and 36 per cent and limb salvage 93 per cent, 85 per cent and 75 per cent at 1, 3 and 5 yr respectively. As a result of poor long term survival (51 per cent at 5 yr), and the healing of remedial lesions before graft occlusion, 68 patients (72 per cent) required no further intervention. Eighteen secondary bypasses were undertaken, 12 using ipsilateral saphenous vein. In this group of elderly patients with poor life expectancy, where a limited operation is desirable, the use of PTFE provided excellent limb salvage with low morbidity. Although we can no longer justify our continued use of PTFE in every case of critical ischaemia because of its inferior patency to autogenous vein, we continue to use it preferentially in patients whose lifespan is likely to be short and in cases where a remedial lesion is present. PMID- 9868859 TI - An unusual cause of otalgia--an ossified stylohyoid ligament. PMID- 9868861 TI - Hepatitis A in urban Ireland. AB - AIMS: To determine the prevalence of immunity to hepatitis A virus (HAV) infection in urban Ireland and to categorize the region into low, intermediate or high HAV endemicity, and to analyse the significance of certain commonly associated risk factors. METHODS: Two hundred and thirty three volunteers were recruited from 6 general practices in Dublin, Ireland. There were 44 volunteers in the 10 to 19 yr age group, 40 in the 20 to 29, 42 in the 30 to 39, 43 in the 40 to 49 and 64 in the over 50 age groups. Each participant completed a detailed questionnaire and was tested for anti-HAV total antibody (primarily IgG) using a competitive ELISA assay. Urban Ireland was classified into the appropriate area of HAV endemicity according to the prevalence of immunity by age group. Risk factor differences were analysed for significance using the chi square test and Fisher's exact test. RESULTS: One hundred and fifty seven (67 per cent) volunteers were immune, of whom 20 (45 per cent) were in the 10 to 19 yr age group, 17 (43 per cent) in the 20 to 29, 30 (71 per cent) in the 30 to 39, 34 (79 per cent) in the 40 to 49 and 59 (92 per cent) in the over 50 age groups. Fifty five per cent of the individuals studied below the age of 20 yr were non-immune. The immune rates over the age of 30 were significantly greater (p < 0.01) than those in the 10 to 29 age groups. Socioeconomic pattern in the total and 10 to 19 yr age group was a significant (p < 0.0002, p < 0.004 respectively) risk factor for infection. CONCLUSION: This study concludes that urban Ireland is an area of low HAV endemicity with age and socioeconomic status as the significant influences on seropositivity. This survey provides an insight into the changing epidemiology of HAV infection in Ireland and serves as a guide for immunisation of at risk population groups. PMID- 9868860 TI - Ret-proto-oncogene analysis in medullary thyroid carcinoma. AB - INTRODUCTION: Medullary carcinoma of the thyroid (MTC) is a rare tumour which occurs in both sporadic and hereditary forms. Mutations of the RET proto-oncogene have been identified in hereditary forms. The aim of our study was to confirm or exclude familial disease by examining for germline mutations in the RET proto oncogene in patients with medullary thyroid carcinoma. METHODS: Nine patients with medullary thyroid carcinoma and 4 of their children were studied. Peripheral blood was used to examine for mutations in the RET proto-oncogene. When this was not available, archival thyroid tissue was used. RESULTS: Seven patients had clinically sporadic tumours confirmed by mutational analysis of RET. Four children were at risk of being carriers of a mutated gene, as their fathers had histologically proven MTC and had tested positive for the mutation at codon 618 on exon 10 of the RET proto-oncogene. Three of these children carried the 618 mutation. To date, 2 have had a prophylactic thyroidectomy, the pathology of which revealed C-cell hyperplasia. One child had familial disease excluded by mutational analysis. One patient had a clinical diagnosis of MEN2B confirmed by detection of the 918 mutation on exon 16 of the RET proto-oncogene. CONCLUSIONS: RET proto-oncogene analysis is a reliable method of differentiating familial from sporadic MTC. Mutational information determines which family members of affected kindreds are at risk of developing the disease and can be used to affect clinical management. PMID- 9868862 TI - Young injectors: a comparative analysis of risk behaviour. AB - This study compares the injecting and sexual risk-behaviour of young injectors, with injectors over the age of 25. All respondents presented for the first time at the Merchants' Quay Health Promotion Unit between May 1st 1997 and February 28th 1998. Analysis revealed that the young injectors were significantly more likely to report recently borrowing and lending used injecting equipment, and injecting paraphernalia. Regarding sexual risk behaviour, younger respondents were proportionately more likely to report being sexually active, having multiple sexual partners, and having a regular partner who is an injecting drug user. However, they were significantly more likely than older clients to report condom use. The suggestion is that the harm minimisation message, which reached its zenith in the early 1990s, has now been somewhat de-emphasised in both policy and practice. Consequently, it is not reaching the young injectors who have recently initiated intravenous drug use. Additional strategies are needed this group of drug users, in order to promote positive behaviour changes. PMID- 9868863 TI - Cardiopulmonary exercise testing in the pre-operative assessment of patients for repair of abdominal aortic aneurysm. AB - We have investigated the value of cardiopulmonary exercise testing in the pre operative assessment to patients for abdominal aortic aneurysm repair. Thirty-six patients were entered into the study. All had a pre-operative clinical assessment and investigations including chest radiograph, electrocardiograph, spirometry and echocardiogram with measurement of left ventricular ejection fraction. Each patient performed a symptom limited treadmill exercise test using a STEEP protocol with on-line measurement of respiratory gas exchange. Patients were followed up for 12 months post-operatively by review of casenotes. Thirty out of 36 patients had surgical repair of abdominal aortic aneurysm. There was 1 death in the perioperative period and 2 deaths in the following 12 months. Seven other patients suffered post-operative complications. There were no significant differences in left ventricular ejection fraction, spirometry and peak achieved oxygen consumption (PVO2) between those patients who died or who had post operative complications and those who had not. However, PVO2 < 20 ml/min/kg was found in 70 per cent of patients who had complications compared with 50 per cent of those who had not. Also 4 patients considered medically unfit for surgery all had PVO2 < 20 ml/min/kg. Cardiopulmonary exercise testing with measurement of PVO2 may be helpful in identifying patients more at risk of post-operative complications but should not be used in isolation without through clinical assessment. PMID- 9868864 TI - Management of the collapsing spine for patients with Duchenne muscular dystrophy. AB - Instrumented fusion of the collapsing spine has gained widespread acceptance for patients with Duchenne Muscular Dystrophy but controversy still exists on the issue of extending the surgical fusion to sacrum in these patients. This retrospective study reviews the long-term outcome of a group of patients with spinal deformity associated with Duchenne Muscular Dystrophy who were managed with long spinal fusion to L5 and ongoing wheelchair seating attention. The clinical notes and radiographs of 19 consecutive patients were reviewed. Fifteen patients attended for clinical and radiological assessment at a mean of 28 months post operatively. The surgery for these patients involved a mean anaesthetic time of 3.5 h and a mean transfusion requirement of 5 units of red cell concentrate. At long-term follow-up 15 patients continued to sit in a well-balanced position. Surgical fusion of the spine to L5 combined with ongoing attention to seating is associated with good long-term functional results in these patients. PMID- 9868865 TI - Establishment and characterization of a hypocatalasemic mouse cell strain. AB - Contact-inhibited catalase-deficient fibroblast cell strain has been established from the homozygous hypocatalasemic C3H/Csb mutant mouse. This cell strain has low level of catalase enzyme activity and has normal level of enzyme activities of both glutathione peroxidase and superoxide dismutase. Catalase-deficient C3H/Csb mutant cell strain is markedly more sensitive to the toxicity of hydrogen peroxide compared to wild-type C3H/Csa cell strain. In addition, mutant cell strain is sensitive to X-rays and near-UV compared to wild-type cell strain, but shows the same sensitivities to topoisomerase II inhibitors, adriamycin and 4'-(9 acridinylamino) methanesulfon-m-anisidide (m-AMSA), and the DNA cross-linking agents, cisdiamminedichloroplatinum (II) (cis-Pt) and trans diamminedichloroplatinum (II) (trans-Pt). These cell strains will be of use in the study of the roles which catalase plays in the intracellular prevention of DNA damage induced by oxidative stress. PMID- 9868866 TI - Increased expression of ornithine decarboxylase by gamma-ray in mouse epidermal cells: relationship with protein kinase C signaling pathway. AB - The correlation between ornithine decarboxylase (ODC) protein induction and specific protein kinase C (PKC) isozyme expression by gamma-ray in 12-O tetradecanoylphorbol-13-acetate (TPA)-treated normal and v-rasHa transformed mouse keratinocytes was examined. TPA at 100 nM was treated in primary mouse keratinocytes immediately after 4 Gy, 8 Gy and 16 Gy gamma-ray irradiation. After 4 hrs, cells were harvested and the protein expression levels of PKC isozymes (PKC alpha, -delta, -epsilon, -eta and -zeta) and ODC were examined. For v-rasHa infection, primary keratinocytes were infected with a defected retrovirus containing the v-rasHa gene. After 3 hrs of irradiation, each PKC isozyme and ODC protein expression were tested. Gamma-ray increases ODC protein expression in both TPA-treated normal and v-rasHa transformed mouse keratinocytes and this phenomenon correlated to the increased induction of PKC alpha without altering other PKC isozymes. Tyrosine phosphorylation of epidermal growth factor receptor protein was also stimulated during gamma-ray induced cellular changes in TPA treated normal mouse keratinocytes. These results indicate that PKC alpha as an important regulator of mouse epidermal changes by gamma-radiation, contributes to the ODC expression occurring during exposure to tumor promoter, such as TPA, and epidermal neoplasia induced by ras activation. PMID- 9868867 TI - Contamination from possible solar light exposures in ESR dosimetry using human tooth enamel. AB - Electron spin resonance (ESR) measurement data of 98 teeth from atomic-bomb survivors who received various radiation doses were analyzed in terms of possible solar light exposure by tooth position. Each tooth was divided into buccal and lingual parts for independent ESR measurement. We found that average buccal doses were larger than their corresponding lingual doses by 0.48 +/- 0.30 Gy (first incisors), 0.33 +/- 0.38 Gy (second incisors), 0.20 +/- 0.23 Gy (canines), 0.24 +/- 0.26 Gy (first bicuspids), 0.17 +/- 0.51 Gy (second bicuspids) and 0.04 +/- 0.18 Gy (large molars and wisdom teeth). Such a clear declining trend following tooth position in the mouth is readily appreciated as due to solar light exposures. Comparisons of lingual doses of multiple teeth from the same five donors suggest that lingual doses of first incisors were also overestimated by 0.34 +/- 0.18 Gy. From the results presented, we deduce that the buccal doses of first incisors are, on the average, overestimated by nearly 0.8 Gy. Therefore, use of front teeth for biodosimetry requires special caution. How to estimate the contribution from exposure to solar light in ESR biodosimetry is discussed. PMID- 9868868 TI - Relative biological effectiveness of accelerated heavy ions for induction of morphological transformation in Syrian hamster embryo cells. AB - Syrian hamster embryo cells were used to study the morphological transformation induced by accelerated heavy ions with different linear energy transfer (LET) ranging from 13 to 400 keV/micron. Exponentially growing cells were irradiated with 12C or 28Si ion beams generated by the Heavy Ion Medical Accelerator in Chiba (HIMAC), then inoculated to culture dishes. Morphologically altered colonies were scored as transformants. Over the LET range examined, the frequency of transformation induced by the heavy ions increased sharply at very low doses no greater than 5 cGy. The relative biological effectiveness (RBE) of the heavy ions relative to X-rays first increased with LET, reached a maximum value of about 7 at 100 keV/micron, then decreased with the further increase of LET. Our findings confirmed that high LET heavy ions are much more effective than X-rays for the induction of in vitro cell transformation. PMID- 9868869 TI - Extension of in vitro life-span of gamma-irradiated human embryo cells accompanied by chromosome instability. AB - We studied the effect of repeated irradiation with a low dose rate (about 0.0012 cGy per min) of 60Co gamma-rays on the in vitro life-span of human embryo (HE) cells. HE cells were cultured in an incubator that was set in a 60Co gamma-ray irradiation room, and the irradiation was repeated throughout the life-span of the HE cells (for about 150 to 160 days) on every day base. During this period, the cells accumulated 106 to 123 cGy. The life-span of the irradiated cells prolonged 1.14 to 1.35 times when compared to that of non-irradiated cells. The incidence of cells with chromosome bridge and micronuclei significantly increased in the irradiated cells. Although the number of chromosomes gradually changed with repetition of culture in both non-irradiated and irradiated cells, the frequencies of aneuploid cells in irradiated cells were about two times higher than that in non-irradiated cells. These results indicate that repeated irradiation with a low dose of gamma-rays produces chromosome instability, and it may be a cause of numerical chromosome abnormalities and life-span extension of irradiated cells. PMID- 9868870 TI - A theoretical model for simultaneous mixed irradiation with multiple types of radiation. AB - Several models for the action of mixed irradiation with two types of radiation have been proposed in the last two decades, although the superiority or inferiority of these models is not completely elucidated. However, mixed irradiation is sometimes composed of more than two types of radiation, and for this type of mixed irradiation, no model has yet been proposed. Therefore, I have tried to make a model for this type of mixed irradiation. This paper presents a survival model for simultaneous mixed irradiation with multiple (n) types of radiation, and this model would be helpful for assessing the effects of simultaneous mixed irradiation with multiple types of radiation in nature, accidents, etc. PMID- 9868871 TI - Estimation of dose absorbed fraction for 131I-beta rays in rat thyroid. AB - The dose absorbed fraction of rat thyroid by internal deposit of 131I has been calculated as a function of effective diameter of thyroid. The calculations were done using two types of Monte Carlo simulations: one was by a simple energy-loss calculation in spherical volume according to the electron stopping power, and another by a more realistic simulation using Monte Carlo N-Particle Transport code system Version 4A (MCNP). These two calculations were consistent with each other within a deviation of 5%. The absorbed fractions in spherical thyroid were drastically changed up to 5 mm diameter, and then almost all energy was deposited within 10 mm diameter. For the practical application to the animal experiment, the absorbed fractions of ellipsoid-shaped thyroids were also calculated for 1-, 4- and 9-week-old rats, where the fractions were estimated to be 0.61, 0.67 and 0.68, respectively. It was also found that the absorbed fraction of the ellipsoid with various dimensions can be simulated by a calculation for spherical volume with a comparable effective diameter. PMID- 9868872 TI - The new focus on spirituality in medicine. PMID- 9868873 TI - At the intersection of clinical and business practices: a catholic perspective. PMID- 9868874 TI - Miracles: quirks of nature or the hand of God. PMID- 9868875 TI - Eckankar: personal experiences with The Holy Spirit. PMID- 9868877 TI - Taoism: the way. PMID- 9868876 TI - Religious congregations as factors in health outcomes. PMID- 9868878 TI - Mormons and health: impact of Latter-day Saint Scriptures on health and health practices. PMID- 9868880 TI - The observant Jewish physician. PMID- 9868879 TI - The evolution of an epidemiologist: a Unitarian-Universalist in public health. PMID- 9868881 TI - Hinduism and medical practice. PMID- 9868882 TI - Jehovah's witnesses and medicine: an overview of beliefs and issues in their care. PMID- 9868884 TI - Flu vaccination decreases relative morbidity risk for people with diabetes. PMID- 9868883 TI - It's about health, it's about time: information everyone should know. PMID- 9868885 TI - A mutual holding company act for Georgians. PMID- 9868886 TI - Surface ultrastructure of Metagonimus miyatai metacercariae and adults. AB - A scanning electron microscopic study was performed to observe surface ultrastructures of excysted metacercariae and adults of Metagonimus miyatai. Metacercariae were collected from the scale of the pale chub (Zacco platypus), and adult flukes were harvested 1-4 weeks after infection to rats. In excysted metacercariae, the oral sucker was devoid of tegumental spines and had type I and type II sensory papillae. Anteriorly to the ventral sucker, spines were dense and digitated into 5-7 points, whereas near the posterior end of the body spines were sparse and digitated into 2-3 points. In one-week adults, 7 type II sensory papillae were arranged around the lip of the oral sucker, and at inner side of the lip one pair of small and two pairs of large type 1 sensory papillae were seen on each side. The distribution of tegumental spines was similar to that of metacercariae, but they were more differentiated with 9-11 pointed tips. In two- to four-week old adults, the surface ultrastructure was nearly the same as in one week old adults, however, sperms were frequently seen entering into the Laurer's canal. Conclusively, the surface ultrastructure of M. miyatai was generally similar to that of M. yokogawai, however, differentiation of tegumental spines and distribution of sensory papillae around the oral sucker were different between the two species, which may be of taxonomic significance. PMID- 9868887 TI - Distribution of actin and tropomyosin in Cryptosporidium muris. AB - Actin and tropomyosin of Cryptosporidium muris were localized by immunogold labeling. Two kinds of antibodies for actin labeling were used. The polyclonal antibody to skeletal muscle (chicken back muscle) actin was labeled on the pellicle and cytoplasmic vacuoles of parasites. The feeder organelle has showed a small amount of polyclonal actin antibody labeling as well. Whereas the monoclonal antibody to smooth muscle (chicken gizzard muscle) actin was chiefly labeled on the filamentous cytoplasm of parasites. The apical portion of host gastric epithelial cell cytoplasm was also labeled by smooth muscle actin together. The polyclonal antibody to tropomyosin was much more labeled at C. muris than host cells, so it could be easily identified even with low magnification (x2,000). The tropomyosin was observed along the pellicle, cytoplasmic vacuoles, and around the nucleus also. The skeletal muscle type actin seems to play a role in various cellular functions with tropomyosin in C. muris; on the other hand, the smooth muscle type actin was located mainly on the filamentous cytoplasm and supported the parasites' firm attachment to host cells. Tropomyosin on the pellicle was thought to be able to stimulate the host as a major antigen through continuous shedding out by the escape of sporozoites or merozoites from their mother cells. PMID- 9868888 TI - Periodicity exhibited by Dirofilaria immitis microfilariae identified in dogs of Korea. AB - Microfilarial periodicity of Dirofilaria immitis (the dog heartworm) was determined at two hr intervals for 72 consecutive hrs in 10 naturally infected war dogs, 3-9 years old, in Korea to facilitate harvest of the microfilariae for possible use in laboratory works and to elucidate further the periodicity of the microfilaria depending on geographic location. Although the periodicity had been observed as being low-grade nocturnal, maximal microfilarial counts were found at 21:00 hr and minimal at 11:00 hr, giving rise to an evident peak in fluctuation of the larval counts. This is the first record of the periodicity of the microfilariae identified as D. immitis in Korea. PMID- 9868889 TI - Current status of vivax malaria among civilians in Korea. AB - A result of national malaria surveillance in Korean civilians was described. Since a case of indigenous vivax malaria was detected in 1993, a total of 2,198 cases was confirmed by blood smear up to 1997. Of them, 1,548 cases were soldiers serving in the demilitarized zone (DMZ), while 650 cases were civilians. Number of civilian cases was 3 in 1994, 19 in 1995, 71 in 1996, and 557 in 1997. Of them, 239 were ex-soldiers who discharged after military service in the prevalent areas such as Paju, Yonchon, Kimpo, Kangwha, Tongduchon in Kyonggi-do and Chorwon in Kangwon-do while 308 patients were civilian residents in the prevalent areas. Seventy-two patients, living nationwide, had a history of visiting the prevalent areas during transmission season. Only 32 civilian patients denied any relation with the prevalent areas. As a whole, a half of the civilian cases was diagnosed when living in non-prevalent areas. Male patients in their twenties was the highest in number. Annual parasite index is steadily elevated in residents living in the prevalent areas. Monthly incidence showed an unimodal distribution, forming a peak in August. Ex-soldiers exhibited a delayed incubation ranging from 153 to 452 days (279 +/- 41 days). The time required for diagnosis was shortened from 23.6 days in 1995 to 13.7 days in 1997. Although the current epidemic of vivax malaria started as a border malaria, it seems highly probable that vivax malaria is established in the local areas and responsible for at least a part of transmission. PMID- 9868890 TI - Control of clonorchiasis by repeated praziquantel treatment and low diagnostic efficacy of sonography. AB - In Korea, Clonorchis sinensis infection is still highly prevalent because case detection in the field is difficult and the detected cases used to be incompletely cured due to treatment failure. The present study tried to control clonorchiasis in an endemic village by repeated treatments with praziquantel every 6 months and to evaluate sonography as a diagnostic measure. By stool examinations, the egg positive rate in the endemic village was 22.7%, but it decreased to 19.6% at 6 months, 15.1% at 12 months. 12.2% at 18 months, 6.3% at 24 months, 11.4% at 30 months, and 6.3% at 42 months after the beginning of repeated praziquantel administration. The sonography showed 61 (49.6%) positive cases of 123 screened residents: among egg-positives the sonography positive rate was 52.2% and among egg-negatives it was still 49%. The rate among cured cases was 64.3% after 6 months, 50.0% after 12 months, 50.0% after 18 months, and 66.7% after 24 months. In a non-endemic village, 64 residents were found egg-negative by fecal examination, but 20 (31.3%) of them were positive by sonography. The present findings indicate that control of clonorchiasis in an endemic village by repeated praziquantel treatment for 42 months is still insufficient and sonography is of little value for diagnosis of clonorchiasis. PMID- 9868891 TI - Biological and biochemical modulation of Trichomonas vaginalis KT9 isolate after shifting of culture medium from TPS-1 into TYM. AB - To evaluate the biological and biochemical characteristics of Trichomonas vaginalis KT9 isolate, the growth and size of trichomonads, pathogenicity in mouse, protein profiles and proteinase activity were examined after shifting the medium from TPS-1 into TYM. Generation time of trichomonads in TYM medium was 4.5 hr in comparison to TPS-1 with 7.1 hr. Size of trichomonads cultured in TPS-1 medium (8.5 +/- 0.9 x 6.0 +/- 0.9 microns) was significantly smaller than those in TYM medium (10.9 +/- 1.4 x 8.2 +/- 0.9 microns). Trichomonads cultured in TYM medium produced subcutaneous abscess in 9 out of 10 mice, whereas those in TPS-1 medium produced abscesses in 2 out of 10 mice. In SDS-PAGE, trichomonad lysates from both media showed ten common bands. However, trichomonads in TYM medium showed additional bands of 136 kDa, 116 kDa and 40 kDa in comparison to those in TPS-1 with 100 kDa. By immunoblot with T. vaginalis-immunized rabbit sera, T. vaginalis cultivated in both TYM and TPS-1 media showed 5 common bands, and unique bands of 116 kDa, 105 kDa, and 86 kDa were observed in trichomonads in TYM while a 140 kDa band in those in TPS-1. In gelatin SDS-PAGE, trichomonads in TYM degraded gelatin stronger than those in TPS-1. Also protease activity of trichomonads in TYM was significantly higher than that of trichomonads in TPS-1 using Bz-Pro-Phe-Arg-Nan as a substrate. According to the results, it is assumed that the shift from TPS-1 into TYM medium for cultivation of T. vaginalis might modulate the biological and biochemical properties of T. vaginalis in vitro. PMID- 9868892 TI - Degradations of human immunoglobulins and hemoglobin by a 60 kDa cysteine proteinase of Trichomonas vaginalis. AB - The present study was undertaken to investigate the role of cysteine proteinase of Trichomonas vaginalis in escaping from host defense mechanism. A cysteine proteinase of T. vaginalis was purified by affinity chromatography and gel filtration. Optimum pH for the purified proteinase activity was 6.0. The proteinase was inhibited by cysteine and serine proteinase inhibitors such as E 64, NEM, IAA, leupeptin, TPCK and TLCK, and also by Hg2+, but not affected by serine-, metallo-, and aspartic proteinase inhibitors such as PMSF, EDTA and pepstatin A. However, it was activated by the cysteine proteinase activator, DTT. The molecular weight of a purified proteinase was 62 kDa on gel filtration and 60 kDa on SDS-PAGE. Interestingly, the purified proteinase was able to degrade serum IgA, secretory IgA, and serum IgG in time- and dose-dependent manners. In addition, the enzyme also degraded hemoglobin in a dose-dependent manner. These results suggest that the acidic cysteine proteinase of T. vaginalis may play a dual role for parasite survival in conferring escape from host humoral defense by degradation of immunoglobulins, and in supplying nutrients to parasites by degradation of hemoglobin. PMID- 9868893 TI - Antibody reaction of human anti-Toxoplasma gondii positive and negative sera with Neospora caninum antigens. AB - Anti-Neospora caninum antibody was detected in anti-Toxoplasma gondii positive and negative human sera by ELISA, western blot and immunofluorescence assay (IFA). Twelve cases out of 172 (6.7%) Toxoplasma-positive sera cross-reacted with both T. gondii and N. caninum antigens, and one out of 110 Toxoplasma-negative sera reacted with N. caninum antigen by ELISA. By western blot, all 12 sera reacted with T. gondii antigens with various banding patterns but specifically at 30 kDa (SAG1) and 22 kDa (SAG2) bands. With N. caninum antigen, the number of reactive bands was reduced, however a 43 kDa band reacted in three cases in Toxoplasma-positive sera in addition to one in Toxoplasma-negative control sera. All sera of the Toxoplasma-positive group labeled surface membrane of T. gondii, but reacted differently with N. caninum. Fluorescence was detected in surface membrane, subcellular organelles, or both in N. caninum. And one case in the Toxoplasma-negative group also reacted with N. caninum strongly in subcellular organelles. This suggested that the antibody against N. caninum may be present in human sera although the positive rate was very low in this study. The possibility of human infection with N. caninum remains to be evaluated further. PMID- 9868894 TI - Karyotype analysis of Neodiplostomum seoulense. AB - A karyotype analysis of the chromosome of Neodiplostomum seoulense, one of causative agents of human intestinal trematodiasis, was done from the gonad tissue by the squashing method. The chromosome number was n = 10 and 2n = 20. Chromosome length was 1.30-4.0 microns. Chromosome pairs in the complement consisted of two pairs of metacentric, five pairs of submetacentrics/subtelocentric and three pairs of telocentric chromosomes. These data were comparable with those of other intestinal trematodes. PMID- 9868895 TI - Verification of immunosuppression in chicks caused by Cryptosporidium baileyi infection using Brucella abortus strain 1119-3. AB - Humoral immune response of young chicks to Brucella abortus strain 1119-3 inoculation was monitored to verify the degree of immunosuppression caused by infection with Cryptosporidium baileyi. Young chicks (2-day-old) were orally inoculated each with 2 x 10(6) oocysts of C. baileyi, and then injected intramuscularly with 0.3 ml B. abortus strain 1119-3 containing 1 x 10(9) living organisms on day 14 postinoculation (PI). Serum samples were tested by plate agglutination test on day 17 PI onwards at an interval of 3-6 days over a period of 36 days. Infected chicks with the coccidium showed significantly lower antibody titers than those of uninfected controls (P < 0.05). These findings document that C. baileyi infection in early life stage may predispose chicks easily to other potential poultry diseases. PMID- 9868896 TI - [A grip grippe?]. PMID- 9868897 TI - [Transdermal therapeutic systems (TTS)]. PMID- 9868898 TI - [Drug allergies]. PMID- 9868899 TI - [How far should one lower LDL cholesterol]. PMID- 9868900 TI - [Changes in sedative therapy?]. PMID- 9868901 TI - [Changes in the concept of multiple sclerosis]. AB - The author summarizes the newest knowledge in the past 10 years that has changed our way of thinking as well as the treatment strategy of the still incurable Multiple Sclerosis. The sophisticated immunological methods have brought new insight into the inflammatory process of the central nervous system. The MR examinations and the newly developed techniques allow examining the dynamism of the disease and the underlying histological alterations. The testing of new drugs -which proved to be efficient--made it possible to follow more precisely not only the classification of the pathological process but the clinical history as well. The new drugs give the patients some hope and the doctors new tools. It enables of slow down the disease progression, so it can reduce the tissue damage caused by the inflammation and can improve the quality of life of the patient until new, more effective therapies are developed. PMID- 9868902 TI - [Multiple endocrine neoplasia-type medullary thyroid carcinoma in three generations of a family]. AB - The familial accumulation of a multiple endocrine neoplasia (MEN) type 2a medullary thyroid carcinoma, is described based on the retrospective analysis of a family history. The proband was characterized by medullary carcinoma (MC) combined with phaeochromocytoma, her child had been shown to suffer from MC. In the third generation of the family C-cell hyperplasia and bilateral adrenal hyperplasia occurred. It is shown that in order to arrive at a correct decision as regards the therapy to be used, modern laboratory tests (serum calcitonin, CEA analysis) and diagnostic imaging methods (ultrasound, computer tomography, magnetic resonance imaging, positron emission tomography, metaiodobenzylguanidine scintigraphy) should be used. It is emphasized that the available therapeutic means (surgery, radiotherapy, nuclear medicine) have to be carefully selected and, if necessary, combined. In medullary thyroid carcinoma associated tumours in other endocrine organs should be expected to occur. Family screening using blood chemical and genetic tests are recommended in asymptomatic cases, since their surgical treatment can in this way lead to complete recovery. PMID- 9868903 TI - [Plasma endotoxin level in healthy blood donors]. AB - The plasma level of endotoxin was determined in 116 healthy blood donors in this laboratory. After a routine physical and laboratory investigations the endotoxin level was determined with Limulus amebocyte lysate assay (LAL test) by the chromogenic kinetic method of Bio-Whittaker Co. (USA). Its sensitivity was 0.005 50 EU/ml. The plasma level of endotoxin in most of healthy donors was less than 1 EU/ml (in the range of 0.01-1.0 EU/ml), but always measurable. The average +/- S. D. was 0.128 +/- 0.215 EU/ml. Because of the high standard deviation and high range of values, the data were distributed to two groups with the means of 0.05 +/- 0.022 EU/ml and 0.294 +/- 0.186 EU/ml. The difference between the groups was significant found (p < 0.001). In conclusion, endotoxin can be measured in plasma of healthy individuals. PMID- 9868904 TI - [A new benzoquinone-containing antimetastatic product]. AB - An orally applicable fermentation product of wheat germ containing 0.04% substituted benzoquinone (MSC) was invented by Hungarian chemists under the trade -name of AVEMAR. The following biological effects of this product were observed. Oral administration (3 g/kg body weight) of MSC enhances blastic transformation of splenic lymphocytes of mice. The same treatment shortens the survival time of skin grafts in co-isogenic mouse skin transplantation model, which points to immune-reconstructive effect of MSC. Highly significant anti-metastatic effect of MSC was observed in three metastasis models (3LL-HH, B16, HCR-25). The antimetastatic activity of MSC--besides the immune reconstitution--may also due to the cell-adhesion inhibitory, cell proliferation inhibitory, apoptosis enhancing and antioxidant effects, which were also observed in our in vitro experiments. Based on the biological effects of MSC--which is non-toxic, according to subacute toxicology studies--this product may be used as an adjuvant in the therapy of malignant neoplasia and other diseases caused by or following immunedeprivation. PMID- 9868906 TI - [Serum alpha-glycoprotein level in gastrointestinal tumors]. PMID- 9868905 TI - [Intrapartum administration of antibiotics in the prevention of neonatal Streptococcus B infections]. AB - This study was performed prospectively. Between 1. January 1995 and 31 December 1997, a modification of the screening-based strategy protocol was implemented. Antenatal screening cultures for GBS were performed at approximately 30-32 weeks of gestation. The protocol recommends the use of antibiotic prophylaxis to GBS positive women with any of the obstetric risk factors for early-onset of GBS disease. Our regiment for prophylaxis for patients in labor was ampicillin 2 g. intravenously then 1 g. i.v. every 4 hours until delivery. Before this study had started (1984-1994), there were 149 serious neonatal GBS infection (149/15,040 pregnancy, among them were 97 premature infants. Thirty-one infants suffered from connatal sepsis. We observed 29 lethal infection. Between January 01, 1995, and December 31, 1997, 4150 women participated in this investigation. The incidence of positive group B Streptococcus cultures from the vaginal samples was 11.6% (481/4150). During the study period (3 years), serious GBS infection was detected in 46 infants (1.1%). There were 9 cases of neonatal sepsis (0.2%), two of them suffered lethal infections (0.05%). There was no late onset of GBS disease and lethal outcome in the last two years. Our investigations bears clinical importance because we confirmed that group B streptococcal colonization is an important risk factor for neonatal infection. The selective intrapartum chemoprophylaxis is a safe and effective intervention to prevent early-onset severe GBS disease. PMID- 9868907 TI - Measles. progress towards global control and regional elimination, 1990-1998. PMID- 9868908 TI - Safety of injections. WHO-UNICEF policy statement for mass immunization campaigns. PMID- 9868909 TI - Low-frequency modulation sensors using nanosecond fluorophores. AB - We describe a new approach to fluorescence sensing based on a mixture of fluorophores, one of which is sensitive to the desired analyte. If a long lifetime analyte-insensitive fluorophore is mixed with a short-lifetime analyte sensitive fluorophore, the modulation of the emission at conveniently low frequencies becomes equal to the fractional fluorescence intensity of the sensing fluorophore. Under these conditions, the modulation can be used to determine the analyte concentration. This can be used with any fluorophore that changes intensity in response to analyte and does not require the sensing fluorophore to display a change in lifetime. The feasibility of modulation-based sensing was demonstrated using mixtures of 6-carboxyfluorescein and [Ru 2,2'-(bipyridyl)3]2+ as a pH sensor and of the calcium probe Fluo-3 and [Ru 2,2'-(bipyridyl)3]2+ as a calcium sensor. PMID- 9868910 TI - Negative ion postsource decay time-of-flight mass spectrometry of peptides containing acidic amino acid residues. AB - Acidic peptides have been studied by negative ion postsource decay (PSD) matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The peptides contained from 5 to 16 residues and were chosen on the basis of their patterns of the acidic residues. Using typical MALDI sample preparation techniques employing an acidic matrix, gastrin I (1-14), and epidermal mitosis inhibiting pentapeptide yielded much larger deprotonated ion signals, [M - H]-, than protonated ions, [M + H]+. This may be due to their absence of basic residues, coupled with their arrays of acidic residues. The PSD fragmentation of the peptide negative ions showed that an array of acidic residues, as in gastrin I (1-14), yielded simple spectra containing mainly backbone cleavage ions from the C-terminus. Hirudin (54-65), which contains two sets of two consecutive Glu residues, and fibrinopeptide A and fibrinopeptide B, with isolated acidic residues, also showed backbone cleavages as common fragment ions. In addition, the two sets of isolated consecutive amino acid residues in Cys(Bzl)84-CD4 (81-92) and hirudin (54-56) yielded internal ions from the cleavages at the (O=C)-NH bond between the acidic residues. Also observed were ions with unique side chain losses, such as the loss of C6H4O from a tyrosine residue and SCH2C6H5 and CH2C6H5 from a benzylated cysteine residue. Compared to the positive mode, the negative-ion PSD yielded fewer fragments which usually involved only one type of backbone cleavage (e.g., [Yn - H2O]-). These simple spectra aided interpretation. Overall, the acidic peptides studied yielded negative ion PSD spectra that were useful for peptide sequencing. PMID- 9868911 TI - Characterization of hemoglobin variants by MALDI-TOF MS using a polyurethane membrane as the sample support. AB - A new method for the sampling and off-site analysis of hemoglobin variants by mass spectrometry is reported. This technique uses a nonporous polyurethane membrane as the collection device and transportation medium of a blood sample for analysis. The same membrane is then used as the MALDI-TOF MS sample support for mass spectrometric analysis. Minimal invasive sample collection is afforded by collecting less than 1 microL of blood using a common lancet device. MALDI-TOF MS is performed directly on the membrane, after washing off the interfering plasma components, followed by the addition of matrix. This reduces the time of analysis and prevents sample loss. Enzymatic digestion can be performed directly on the membrane, using in this case trypsin, allowing for further characterization of the sample. The method is much less invasive compared to drawing blood with a syringe. The sample may be transported to the laboratory by regular mail, and thus the method can serve remote locations. We demonstrate the procedure by characterizing the Hb Shepherds Bush hemoglobin variant, b74-(E18)Gly-->Asp. PMID- 9868912 TI - Modification of cysteine residues by alkylation. A tool in peptide mapping and protein identification. AB - Although mass spectrometric peptide mapping has become an established technique for the rapid identification of proteins isolated by polyacrylamide gel electrophoresis (PAGE), the results of the identification procedure can sometimes be ambiguous. Such ambiguities become increasingly prevalent for proteins isolated as mixtures or when only very small amounts of the proteins are isolated. The quality of the identification procedure can be improved by increasing the number of peptides that are extracted from the gel. Here we show that cysteine alkylation is required to ensure maximal coverage in matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) peptide mapping of proteins isolated by PAGE. In the described procedure, alkylation was performed prior to electrophoresis to avoid the adventitious formation of acrylamide adducts during electrophoresis. In this way, homogeneous alkylation was obtained with three different alkylating reagents (4 vinylpyridine, iodoacetamide, acrylamide). Cysteine alkylation was also used as a tool for the identification of cysteine-containing peptides. Using a 1:1 mixture of unlabeled acrylamide and deuterium-labeled acrylamide ([2,3,3'-D3]acrylamide), the proteins of interest were alkylated prior to electrophoretic separation. Peptide mixtures produced by trypsin digestion of the resulting protein bands were analyzed by MALDI-TOF MS, and the cysteine content of the peptides was inferred from the isotopic distributions. The cysteine content information was readily obtained and used to improve the protein identification process. PMID- 9868913 TI - C-terminal peptide sequencing via multistage mass spectrometry. AB - Results are presented showing the ability to obtain C-terminal sequence information from peptides by multiple stages of mass spectrometry. Under typical low-energy collision-induced dissociation conditions of quadrupole ion trap and ion cyclotron resonance mass spectrometers, lithium- and sodium-cationized peptides dissociate predominantly by reaction at the C-terminal peptide bond or an adjacent bond. For the majority of cases studied, the dominant reaction is a rearrangement process that results in the loss of the C-terminal residue and formation of a product ion that is one amino acid shorter than the original peptide ion. Using the multistage MS/MS capabilities of quadrupole ion trap and ion cyclotron resonance mass spectrometers, a subsequent stage of MS/MS can be performed to determine the identity of the new C-terminal residue. Up to eight stage of MS/MS have been performed with both quadrupole ion trap and ion cyclotron resonance mass spectrometers. In general, the same dissociation pathways are observed with both instruments, although occasionally there are significant differences in the branching ratios of competing pathways. PMID- 9868914 TI - Enantiomeric separations of terbutaline by CE with a sulfated beta-cyclodextrin chiral selector: a quantitative binding study. AB - Sulfated beta-cyclodextrin, a negatively charged chiral selector, was used for the enantiomeric separation of racemic terbutaline by capillary electrophoresis. Chiral separation was found to increase with decreasing cyclodextrin concentration. Host-guest complex binding constants for this system were determined by UV difference spectroscopy (Kav = 1490 M-1) and by CE under conditions of minimal EOF and reversed polarity (K1 = 1730 M-1, K2 = 1590 M-1, alpha = 1.09). The effect of organic modifiers, methanol, and acetonitrile was also studied over a wide range of modifier concentrations. Binding constants decreased while selectivity increased with increasing organic modifier concentration (10% MeOH: K1 = 1590 M-1, K2 = 1130 M-1, alpha = 1.41. 10% ACN: K1 = 1320 M-1, K2 = 870 M-1, alpha = 1.52). Experimental results are discussed in the context of existing separation models. PMID- 9868915 TI - Multiple sample PCR amplification and electrophoretic analysis on a microchip. AB - Polymerase chain reactions (PCRs) were carried out on as many as four DNA samples at a time on a microchip device. The PCR products were then analyzed, either individually or together on the same device, by microchip gel electrophoresis. A standard PCR protocol was used to amplify 199- and 500-base pair (bp) regions of bacteriophage lambda DNA and 346- and 410-bp regions of E. coli genomic and plasmid DNAs, respectively. Thermal lysis of the bacteria was integrated into the PCR cycle. A product sizing medium, poly(dimethylacrylamide), and an intercalating dye for fluorescence detection were used in the electrophoretic analysis of the products. PCR product sizes were determined by coelectrophoresis with marker DNA. PMID- 9868916 TI - Colloidal Au-enhanced surface plasmon resonance immunosensing. AB - Surface plasmon resonance (SPR) biosensing using colloidal Au enhancement is reported. Immobilization of approximately 11-nm-diameter colloidal Au to an evaporated Au film results in a large shift in plasmon angle, a broadened plasmon resonance, and an increase in minimum reflectance. The incorporation of colloidal Au into SPR biosensing results in increased SPR sensitivity to protein-protein interactions when a Au film-immobilized antibody and an antigen-colloidal Au conjugate comprise the binding pair. A highly specific particle-enhanced analogue of a sandwich immunoassay is also demonstrated by complexing the Au particle to a secondary antibody. A tremendous signal amplification is observed, as addition of the antibody-Au colloid conjugate results in a 25-fold larger signal than that due to addition of a free antibody solution that is 6 orders of magnitude more concentrated. Picomolar detection of human immunoglobulin G has been realized using particle enhancement, with the theoretical limits for the technique being much lower. Finally, a quasi-linear relationship between particle coverage and plasmon angle shift is presented, thereby providing for a direct correlation between plasmon shift and solution antigen concentration. Together, these results represent significant advances in the generality and sensitivity of SPR as it is applied to biosensing. PMID- 9868917 TI - Stable cationic capillary coating with successive multiple ionic polymer layers for capillary electrophoresis. AB - A coated capillary modified with a cationic polymer was developed by using a novel coating procedure, successive multiple ionic-polymer (SMIL) coating. The SMIL coating was achieved by first attaching the cationic polymer to the capillary inner wall, and then the anionic polymer to the cationic polymer layer, and finally the cationic polymer to the anionic polymer layer. The stability of Polybrene (PB)-modified capillary made by SMIL coating was remarkably improved in comparison with a conventional PB-modified capillary. It endured during 600 replicate analyses and also showed strong stability against 1 M NaOH and 0.1 M HCl. The relative standard deviation of the run-to-run, day-to-day, and capillary to-capillary coating was all below 1%, and good reproducibilities were obtained. The PB-modified capillary made by SMIL coating was applied to the basic protein analyses. It gave good performances for the protein analyses even when the pH of the electrolyte was near the isoelectric point (pI) of the protein. In addition, 0.1 M NaOH rinse prior to the sample injection allowed the reproducible analysis of a highly adsorptive sample such as plasma because the adsorbed sample could be flushed out of the capillary. Besides protein analyses, an efficient analysis of the cationic drugs by capillary electrophoresis/mass spectrometry (CE/MS) was also possible. PMID- 9868918 TI - On-line MALDI-TOF MS using a continuous vacuum deposition interface. AB - In this work, a new interface for continuous on-line MALDI-TOF MS is presented. The sample, mixed with a suitable matrix, was transported into the evacuated source chamber of the mass spectrometer at liquid flow rates of 100-400 nL/min. The liquid sample matrix was deposited on a rotating quartz wheel and transported to the repeller, where laser desorption took place. Rapid evaporation of the solvent (water or methanol) on the surface of the wheel resulted in formation of a thin, approximately 50-micron-wide, sample trace. Scanning electron microscopic photographs of the vacuum-dried trace revealed the deposited material to consist of an amorphous film. Furthermore, sample uniformity along the trace, in conjunction with its narrow width, resulted in excellent signal reproducibility, with detection limits in the attomole range. The interface permitted the on-line coupling of microcolumn separation techniques with MALDI MS, as demonstrated in the capillary electrophoresis MALDI-TOF MS analysis of a 12-peptide mixture. The approach offers the potential for rapid separation and trace analysis of complex mixtures. PMID- 9868919 TI - On-line cation exchange for suppression of adduct formation in negative-ion electrospray mass spectrometry of nucleic acids. AB - One major difficulty in the analysis of nucleic acids by electrospray mass spectrometry is represented by the affinity of the polyanionic sugar-phosphate backbone for nonvolatile cations, especially ubiquitous sodium and potassium ions. A simple on-line sample preparation system comprising a microflow pumping system and 45 x 0.8-mm-i.d. microcolumns packed with weak or strong cation exchange resins is described for the efficient removal of cations from nucleic acid samples. Samples were analyzed by flow injection analysis at a 3-5 microL/min flow of 10 mM triethylamine in 50% water-50% acetonitrile. After on line desalting, mass spectra of oligonucleotides revealed no significant sodium adduct peaks. Moreover, signal-to-noise ratios were greatly enhanced compared to direct injection of the samples. Electrospray mass spectrometry with on-line sample preparation allowed accurate molecular mass determinations of picomole amounts of crude oligonucleotide preparations ranging in size from 8 to 80 nucleotides within a few minutes. The good linearity of the calibration plot (R2 = 0.9988) over at least 2 orders of magnitude and a relative standard deviation in peak areas of less than 9% permitted the sensitive quantitative measurement of oligonucleotides in a concentration range of 0.2-20 microM with selected-ion monitoring. Finally, the on-line sample preparation system was evaluated for the mass spectrometric analysis of complex oligonucleotide mixtures. PMID- 9868920 TI - Estimation of bacterial contamination in ultrapure water: application of the anti DNA antibody. AB - We constructed and established a hybridoma cell line that produces immunoglobulin G-type anti-DNA antibody. By using this antibody, we could successfully detect a single bacterial cell in ultrapure water (UPW). The detection system is composed of a membrane-supported western blotting-type immunoassay and a two-dimensional photon analyzer with high resolution. It can detect and count every bacterial cell in a wide field of view on a trapping filter i.e., a circle with an 18-mm diameter. This means 10 fg (10(-14) g) of bacterial DNA can be detected in the field. This system could be a useful tool for evaluating the number of bacteria contained by UPW and water used for medical purposes. PMID- 9868921 TI - Limited-sample NMR using solenoidal microcoils, perfluorocarbon plugs, and capillary spinning. AB - This study demonstrates three improvements to mass-limited NMR using solenoidal microcoils as detectors: (1) sample confinement using liquid perfluorocarbon plugs to increase the observe factor, (2) design and incorporation of a capillary spinner to improve spectral line widths, and (3) facile sample changing via the use of a capillary insert. The probe is constructed to spin a fused silica capillary of 530 microns i.d., 700 microns o.d. inside a solenoidal coil wound around a 0.8 mm i.d., 1 mm o.d. glass capillary. The smaller capillary contains the sample, and capillaries with different samples can be exchanged easily. In high-resolution limited sample microcoil NMR studies published thus far, the length of the sample plug has been 7-10 times the length of the solenoid to avoid line broadening from volume magnetic susceptibility (chi v) mismatches at both ends of the sample. This arrangement is not efficient since it places most of the sample volume outside of the coil observe volume. It is shown here that the observe factor cannot exceed 23% if the sample plug is bracketed by air, without substantial line broadening occurring. However, if the sample is bracketed by two liquid perfluorocarbon plugs, the observe factor can be increased to 70% while maintaining high spectral resolution. This is equivalent to improving the limit of detection by a factor of 3, or reducing the total data acquisition time for a given signal-to-noise by a factor of 9. It is also shown that, for the 440-nL sample plug used in this study (bracketed by the perflurocarbon plugs), sample spinning can improve the spectral resolution from 1.5 (nonspinning) to 0.6 Hz (spinning). This corresponds to a further improvement in the limit of detection of 2.5, or just over a factor of 6 decrease in data acquisition time. PMID- 9868922 TI - Precolumn affinity capillary electrophoresis for the identification of clinically relevant proteins in human serum: application to human cardiac troponin I. AB - An approach has been developed to the on-line extraction and identification of clinical disease-state marker proteins in human serum. Fabrication of capillaries with integral packed beds for the online determination of human cardiac troponin I (cTnI), a diagnostic marker for myocardial infarction, at clinically relevant levels (2 nmol/L) in serum is demonstrated. The technique, termed precolumn affinity capillary electrophoresis (PA-CE), utilizes a short (approximately 5 mm) packed bed of porous silica containing covalently immobilized monoclonal anti cTnI antibodies directly integrated within a separation capillary for the selective retention of cTnI from a complex matrix. Following a rinsing step to eliminate nonspecifically bound serum proteins and other impurities from the column, desorption of the antigen into the separation region of the PA-CE capillary for subsequent measurement of femto-molar amounts of cTnI by CE is effected by the injection of an appropriate elution buffer. Advantages of this approach over previously reported affinity preconcentration techniques, related applications for PA-CE technology, and its potential for use in the development of a certified reference material for cTnI in serum are discussed. PMID- 9868923 TI - Nanoliter chemistry combined with mass spectrometry for peptide mapping of proteins from single mammalian cell lysates. AB - A nanoliter-chemistry station combined with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was developed to characterize proteins at the attomole level. Chemical reactions including protein digestion were carried out in nanoliter or subnanoliter volumes, followed by microspot sample deposition of the digest to a MALDI-TOF mass spectrometer. Accurate mass determination of the peptides from the enzyme digest, in conjunction with protein database searching, allowed the identification of the proteins in the protein database. This method is particularly useful for handling small-volume samples such as in single-cell analysis. The high sensitivity and specificity of this method were demonstrated by peptide mapping and identifying hemoglobin variants of sickle cell disease from a single red blood cell. The approach of combining nanoliter chemistry with highly sensitive mass spectrometric analysis should find general use in characterizing proteins from biological systems where only a limited amount of material is available for interrogation. PMID- 9868924 TI - Cytogenetic characterization of an extra structurally abnormal chromosome associated with severe mental retardation: inv dup (15) (q13). AB - We have studied an extra structually abnormal chromosome (ESAC) in a 13 years old boy with profound mental, psychomotor and speech retardation, behavioral problems, seizures and abnormal electroencephalogram. The examination of the bisatellited ESAC with chromosome banding demonstrated that the karyotype was: 47, XY, +inv dup (15) (pter-->q13::q13-->pter). The cytogenetic characterization of the inv dup (15) is reported with special emphasis on the usefulness of DA/DAPI staining when G-banding is sequentially performed to discard possible heteromorphisms in DA/DAPI positive chromosomes, and the importance of Ag-NOR heteromorphisms to ascertain the maternal origin of the inv dup (15). A U-type exchange between two non-sister chromatids is proposed as its mechanism of formation. The clinical features of the case were consistent with those previously reported in similar cases. PMID- 9868925 TI - Larval tolerance in the Drosophila melanogaster species complex toward the two toxic acids of the D. sechellia host plant. AB - The toxicity of hexanoic (C6) and octanoic (C8) acids, the two major components of the host plant of Drosophila sechellia, was investigated upon larvae of the four species included in the D. melanogaster complex and on interspecific hybrids between D. sechellia and D. simulans. Specific methods had to be devised for obtaining reproducible toxicity results. The three generalist species (D. melanogaster, D. mauritiana and D. simulans) were found to be very sensitive, as indicated by low lethal concentrations and an increase in development duration. By contrast D. sechellia was much more tolerant, especially toward C8 which is the most abundant product in the natural resource. Interspecific hybrids (F1 and backcrosses) exhibited intermediate characteristic, but a dominance of D. simulans sensitivity was observed for both acids and especially for C8. Data on larvae are quite different from those previously obtained on adults, and are more likely to reflect the natural selective pressures existing in the wild. PMID- 9868926 TI - Microsatellite variation in Scandinavian Cervidae using primers derived from Bovidae. AB - The possibility of using microsatellite primers developed in Bovidae to amplify microsatellite markers in Cervidae was surveyed by using 75 microsatellite primer sets of bovine, ovine or caprine origin to analyse DNA from moose, red deer, reindeer and roe deer from Scandinavia. On average for the four cervids, approximately 50% of the ovine/caprine primer pairs amplified a specific PCR product, compared to only 16% of the bovine primers. Approximately 50% of both ovine/caprine and bovine primers that amplified a specific product were polymorphic, giving 15 polymorphic microsatellite markers in moose, 11 in red deer, 21 in reindeer and 10 in roe deer. Reindeer had a higher proportion of polymorphic loci, more alleles per locus and higher mean heterozygosity than the other cervids. PMID- 9868927 TI - Mitochondrial cytochrome b DNA sequence variation of Atlantic cod, Gadus morhua, from the Baltic and the White Seas. AB - We studied sequence variation of a fragment of the mitochondrial cytochrome b gene using polymerase chain reaction (PCR) and direct sequencing of 109 Atlantic cod (Gadus morhua) from the Baltic and 19 cod from the White Sea. Eight nucleotide substitutions define 12 haplotypes differing from each other by one to six substitutions. All mutations are at silent sites and thus neutral to selection arising from protein action. Frequencies of four common haplotypes are 6%-57% and haplotype diversity is 0.64 +/- 0.031. Testing for neutral theory predictions reveals no departure from neutrality. The Baltic shares its haplotypes with distant localities in the Atlantic but differs in frequency. Within the Baltic none of the variation is explained by geographic location with an AMOVA. Temporal changes of haplotype frequencies of up to 50% are observed between sampling years and yearclasses. A potential confounding of temporal and microspatial variation cannot be excluded due to limited sample size. PMID- 9868928 TI - Nucleoli, rRNA genes and ITS region in Posidonia oceanica (L.) Delile. AB - The maximum number of nucleoli was counted in interphase nuclei of Posidonia oceanica, and a restriction pattern of nuclear rDNA was obtained after digestion with four restriction endonucleases and Southern hybridization. P. oceanica has only one type of ribosomal gene whose size was estimated to be 18.5 kbp long. The nucleotide sequence of the entire ITS region was also determined by direct sequencing of PCR amplified DNA fragments. The sequence of the ITS region was aligned with those of homologous regions of other monocots available in literature, and phylogenetic trees were obtained. PMID- 9868929 TI - Evaluation of genetic variability in introduced populations of red deer (Cervus elaphus) using DNA fingerprinting. PMID- 9868930 TI - Karyotype analysis of interspecific rat/mouse somatic cell hybrids by reverse chromosome painting. PMID- 9868931 TI - A maize MuDR-like element expressed in rice callus subcultured with proline. PMID- 9868932 TI - Sealing ability of composite resin placed over calcium hydroxide and calcium sulphate plugs in the repair of furcation perforations in mandibular molars: a study in vitro. AB - The purpose of this study was to evaluate in vitro the sealing ability of various materials in the repair of furcation perforations in mandibular molars by measuring coronal microleakage with Indian ink. Ninety extracted mandibular molars were embedded individually into a plaster of Paris block, with the roots surrounded by a simulated periodontal ligament of silicone. Subsequently, a standard coronal access opening was prepared, the root canal orifices were located and a perforation was made with a size 012 round bur in a water-cooled high-speed handpiece directly into the centre of the floor of the pulp chamber. The perforations were repaired with amalgam, composite resin, calcium sulphate under composite resin and calcium hydroxide under composite resin. The teeth were coated with two layers of nail polish, leaving the access opening area uncovered, and immersed in Indian ink for 4 days at 37 degrees C. The teeth were sectioned longitudinally and dye penetration measured from the coronal level of the repair material to the apical end of the perforation. All experimental groups revealed dye penetration in varying degrees, but there was no significant difference amongst them (Kruskal-Wallis test P < 0.05). Calcium sulphate and calcium hydroxide prevented overextrusion of composite resin when used under this repair material. PMID- 9868933 TI - Effect of different root canal sealers on periapical repair of teeth with chronic periradicular periodontitis. AB - Teeth with induced chronic periradicular periodontitis in dogs were root canal treated. After the biomechanical preparation, using K files and 5.25% sodium hypochlorite as the irrigant solution, all root canals were dressed with an antibacterial dressing based on calcium hydroxide, which was left in place for 7 days. After this time, the root canals were obturated with lateral condensation of cold gutta-percha with either a calcium hydroxide root canal filling material (Sealapex) or a zinc oxide-eugenol sealer (Fill Canal). After 270 days, histopathological analysis showed better apical and periapical repair in the teeth obturated with Sealapex (P < 0.05). PMID- 9868935 TI - Influence of file handle attachments on tactile discrimination: an evaluation by dental students. AB - This study used a simulator to determine the effectiveness of two file-handle sleeves that are claimed to improve tactile discrimination and the ease of manipulation of endodontic hand instruments. Thirty final-year dental undergraduates inserted files into instrument setting rods until a putty resistance was felt, following which depth of penetration into the material was measured with a microscope. Results were analysed statistically. Despite favourable comments from participants in the trial, the devices failed to deliver enhanced tactile discrimination. PMID- 9868934 TI - In vitro dentinal penetration by tracers used in microleakage studies. AB - Many microleakage studies, in their methodology, employ different tracers which may penetrate around restorative materials to varying extents due to their physical or chemical characteristics. The present series of in vitro studies were designed to compare the ability of freshly obtained microleakage tracers to enter cut dentinal tubules. Tracers selected were 5% eosin, 2% methylene blue, 50% silver nitrate and Indian ink all buffered, where necessary, to a range of pH 6.9 7.2. The solution of Indian ink was further investigated by particle size analysis to determine the range of particle sizes within the solution. Particles ranged from < 1 micron to 600 microns. Buffered and unbuffered solutions of the tracers were applied for 1 h to open cavities, devoid of a smear layer, prepared in 42 premolars. Specimens were sectioned longitudinally through the cavities either with or without water coolant. Penetration into dentinal tubules by the tracer resulted in an area of stained dentine and this area was subsequently measured using image analysis. The results demonstrated that there were few statistically significant differences between the areas of dentinal penetration associated with each tracer. Neither the tracer used, its pH nor sectioning technique had a predictably significant effect on dentinal penetration. Indian ink was observed to be capable of entering dentinal tubules. PMID- 9868936 TI - Preparation of the apical part of the root canal by the Lightspeed and step-back techniques. AB - This study measured in vitro the displacement of natural canal centres in 18 human teeth before and after shaping by the step-back or Lightspeed techniques. Experimental roots (n = 9 per group), embedded in clear plastic, were cross sectioned using a 0.1-mm-thick band saw at distances 1.25 mm, 3.25 mm and 5.25 mm from the apices. A stereo microscope was used to take 35 mm slides of the cut surfaces of the sectioned roots and canals. The slides of the uninstrumented canals were scanned into a computer and saved. Each sectioned root was then reassembled and the canals shaped by the step-back or Lightspeed technique. File size 40 and instrument size 50 were selected as the master apical file and master apical rotary for the step-back and Lightspeed groups, respectively. The 18 prepared canals were photographed, and the 35 mm slides scanned and computer stored as previously. This allowed the positions of the pre- and postinstrumented roots to be electronically superimposed for subsequent analyses. Displacements of the root canal centres before and after preparation were assessed in relation to the cross-sectional diameter of the files or instruments used. In addition, increases in cross-sectional area of the root canals after preparation were evaluated in relation to the cross-sectional area of the files or instruments used. Engine-driven nickel-titanium Lightspeed instruments caused significantly less (P < 0.001) displacement of the canal centres, so roots in the Lightspeed group remained better centred than those in the step-back group. The mean cross sectional area after preparation in the Lightspeed group was significantly less (P < 0.001) than that recorded in the step-back group. Clinically, this implies less apical transportation and less dentine destruction with the Lightspeed technique than with the step-back technique. PMID- 9868937 TI - Effect in vitro of Tifell (formocresol-eugenol) on macrophage adhesion. AB - The purpose of this study was to investigate the effect in vitro on macrophage adhesion of Tifell (formocresoleugenol), used as an intracanal medicament and in therapeutic pulpotomies in primary teeth, Macrophages were obtained from Wistar rats. As a test of macrophage phagocytic function the adherence capacity of macrophages to a plastic surface was determined. Assays were carried out in Eppendorf tubes after 15 min of incubation at 37 degrees C in a humidified atmosphere of 5% carbon dioxide. The adherence index (AI) was calculated. Results showed that Tifell decreased significantly (P < 0.05) the AI of macrophages. Half maximal inhibition of AI was obtained at 1:334.5 Tifell dilution (AI = 27.75; P < 0.05). Taking into account that substrate adherence is the first step in the phagocytic action of macrophages and in antigen presentation, Tifell could inhibit macrophage function and modulate immune and inflammatory responses in dental pulp and periapical tissues. PMID- 9868938 TI - Study of the number of roots and canals in Senegalese first permanent mandibular molars. AB - A study of 480 extracted first permanent mandibular molars of the Oulof people of Senegal, West Africa, revealed 15 teeth with three roots (3.12%), and 120 teeth with four root canals (25%). An extra root correlated with a sextum lingual tubercle in 20% of cases. Analysis of the pulp chambers revealed a variety of rectangular shapes. Comparison of these anomalies with their incidence in other ethnic groups extends the range of incidence of three-rooted first permanent mandibular molars in the Negroid race. This trait is described for only the second time in a Negroid population. The clinical consequences of root variation in root canal treatment are considered. PMID- 9868939 TI - A comparison of digitally scanned radiographs with conventional film for the detection of small endodontic instruments. AB - The use of computers in dentistry is becoming common as a practice tool for a diverse number of tasks, including the storage and enhancement of intra-oral radiographs. Several systems of digital radiography are available to produce a digital image including irradiation of a charged-couple device and scanning conventional radiographs. This study compared various digital images of scanned periapical radiographs with the original radiographs to determine whether the digitized images offered any advantage when viewing small files at the radiographic apex. Twenty extracted permanent molar teeth were prepared by gaining straight line access to the root canals and a ISO size 06 K-file was introduced into one of the canals until the tip was flush with the apical foramen. Using a standardized technique, radiographs were taken of the teeth using E-speed film. The radiographs were scanned and five digital images: original, enhanced, negative to positive conversion, zoom and zoom of negative to positive were produced. Three evaluators compared each of the images with the radiograph for clarity of the endodontic file in relation to the radiographic apex. Results were analysed using the Wilcoxon signed rank test and the Kappa (kappa) test was used to measure the level of agreement between the three evaluators. The results revealed that all the digital images produced by this scanner were inferior to the radiograph (P < 0.001) and that there was high agreement between evaluators. PMID- 9868940 TI - Vertical root fractures in adjacent maxillary premolars: an endodontic-prosthetic perplexity. AB - Vertical root fractures in endodontically treated teeth occur frequently in teeth or roots in which their mesiodistal dimension is narrow, such as the maxillary upper premolars. Two cases of vertical root fractures in two adjacent maxillary reconstructed teeth are presented. As in many cases of vertical root fractures, the primary diagnosis was of endodontic treatment failure. The final diagnosis of the fractures was made either by the radiograph showing separation of root segments in one case, or by surgical exploration in the other. In both cases, tooth extraction was unavoidable. PMID- 9868941 TI - A teaching model for endodontic surgery. AB - It is often difficult to achieve realistic simulation in teaching endodontic surgery. There is relatively little material available to students other than animal tissue or human cadavers. We propose a working model for the teaching of this discipline, constructed from casts of a natural skull which reproduce anatomical features such as gingiva, maxillary sinus, or mandibular canal, for example. This model, made of polyurethane resin containing mineral particles, which closely simulates the radiological and tactile differences of the relative densities of spongy bone, cortical bone and teeth, permits close simulation of the conditions under which endodontic surgery may be performed. PMID- 9868942 TI - The influence of storage conditions on the clonogenic capacity of periodontal ligament cells: implications for tooth replantation. AB - Viable periodontal ligament (PL) cells are required for PL healing of avulsed teeth following replantation. If immediate replantation cannot be accomplished, the ability of PL progenitor cells to reproduce (clonogenic capacity) and recolonize the wound may be extended by prevention of desiccation and storage in physiological media. This investigation examined the effects of storage in saliva, milk, Hank's balanced salt solution (HBSS) and Eagle's medium (alpha MEM) on the clonogenic capacity of human PL progenitor cells at 30 and 60 min extra alveolar time. Twenty erupted human premolar teeth extracted as atraumatically as possible for orthodontic purposes were used in the present study. Fifteen premolars were placed immediately in freshly collected autologous saliva at room temperature, (+ 23 degrees C) for 15 min. These 15 premolars were next divided into three groups of five and stored in either saliva, milk or HBSS at + 4 degrees C in plastic cups surrounded by ice. The remaining five teeth served as positive controls and were immediately placed in alpha MEM at + 4 degrees C. PL tissue was scraped from one-half of the root surface with a scalpel at 30 and 60 min total extra-alveolar duration. Cells were released from the tissue sample with a 30 min enzymatic digestion procedure and the cells from the tissue samples analyzed for clonogenic capacity. There was a reduction in clonogenic capacity with time for all protocols. Periodontal ligament cells stored in alpha MEM showed the least reduction between 30 and 60 min and the greatest reduction was observed for PL cells stored in saliva. The difference in clonogenic capacity following transfer from saliva to milk or HBSS was not significant at 30 min. At 60 min, cells transferred from saliva to HBSS had a statistically higher percentage of clonogenic cells than those transferred to milk (5.9% vs. 3.5%; P < 0.05). We conclude that immediate storage of avulsed teeth in autologous saliva, followed by transfer to chilled milk, preserves the presence of sufficient progenitor cells in the PL to warrant replantation and the possibility of PL healing at 60 min extra-alveolar duration. PMID- 9868943 TI - Degradation of a silver point in association with endodontic infection. AB - This paper describes an unusual clinical report of degradation of an endodontic silver point within the root canal of a lower right first premolar. Its apical portion was found to be degrading and at one stage a small portion became separated from its main body and was displaced apically. The tooth, despite past apical surgery, displayed signs of endodontic failure. Conventional retreatment and removal of the silver point improved the endodontic problems associated with this tooth. The silver point was analysed using both scanning electron microscopy and x-ray microanalyses. These tests revealed that the silver point had deteriorated significantly with a surface coating of a silver-chloride salt. Microleakage with infection and galvanic reaction are possible hypotheses as to the reason for the degradation of the silver point. Although other reports have demonstrated corrosion of silver points, this method of presentation appears unique. PMID- 9868944 TI - Normal chest radiograph and lung function do not necessarily mean normal lungs. PMID- 9868945 TI - Surgical treatment of Takayasu's Arteritis. PMID- 9868946 TI - Partial hypoxanthine phosphoribosyltransferase deficiency: unrecognized until adult ages. PMID- 9868947 TI - Hyponatremia: how to approach this confusing abnormality. PMID- 9868949 TI - Hypophosphatemia and encephalopathy in alcoholics. PMID- 9868948 TI - Many faces of mixed medullary-follicular and papillary carcinoma of the thyroid. PMID- 9868950 TI - Association of chronic infection of Chlamydia pneumoniae and coronary heart disease in the Japanese. AB - The association of Chlamydia pneumoniae with atherosclerosis of coronary and carotid arteries and the aorta has been demonstrated by seroepidemiology and by detection of the organism in atheromata. We investigated the frequency of C. pneumoniae seropositivity in patients with acute myocardial infarction (AMI). C. pneumoniae-specific antibodies were measured by the microimmunofluorescence test in 160 AMI patients and 160 control subjects matched for age and gender. The odds ratios (ORs) were 2.2 (95% confidence interval (CI), 1.2 to 3.9) for immunoglobulin (Ig)G and 2.7 (95% CI, 1.7 to 4.3) for IgA. After adjustment for other cardiovascular risk factors of age, gender, hypertension, diabetes, cigarette smoking and serum cholesterol, the ORs were essentially unchanged. This study confirmed that the observations of an association between antibody against C. pneumoniae and coronary heart disease in Western nations is also present in Japan. Our results are comparable to the previous seroepidemiological studies reporting ORs of 2.0 or greater. PMID- 9868951 TI - Mechanism of increased serum cytokeratin 19 fragment levels in patients with diabetic nephropathy as a model of chronic renal failure. AB - We examined serum and urinary cytokeratin 19 fragment (CYFRA 21-1) levels in patients with diabetic nephropathy as a model of chronic renal failure, to investigate the mechanism of increased serum CYFRA 21-1 levels in chronic renal failure. Serum and urinary CYFRA 21-1 levels in non-insulin-dependent diabetes mellitus (NIDDM) patients with abnormal urinary immunoglobulin G (IgG) levels (>1.1 mg/g x Cr, n=126) were higher than those with normal urinary IgG levels. In NIDDM patients with normal urinary IgG levels (n=81); the urinary albumin or transferrin levels were not related to serum or urinary CYFRA 21-1 levels. We speculate that the increased serum CYFRA 21-1 levels contribute to metabolic abnormality in the kidney itself rather than the decreased urinary excretion per se, and that increased urinary CYFRA 21-1 levels are found in advanced cases of diabetic nephropathy with destruction of the size barrier. PMID- 9868952 TI - Pulmonary manifestations in cerebrotendinous xanthomatosis. AB - We investigated five cases with cerebrotendinous xanthomatosis (CTX) with particular reference to biochemical and pathological pulmonary disorders. To date, few reports discuss the pathophysiology of pulmonary disorders of CTX patients. This study is the first investigation of such pulmonary disorders. All 5 patients had no pulmonary symptoms and no disturbances on radiological studies and pulmonary function tests. However, in bronchoalveolar lavage (BAL) fluids, many cells with cruciform reflexes, which is characteristic of intracellular sterol accumulation, were found under phase contrast microscopy. Biochemically, cholestanol was found to be increased in the BAL fluid as well as in serum. Pathological findings of transbronchial lung biopsy (TBLB) samples disclosed foamy macrophages and small granulomas in alveolar septa. In conclusion, the lung was apparently involved in CTX, and the lesions were characterized with the accumulation of foamy and giant cells with a high concentration of cholestanol, which likely results in the formation of foreign body granulomas. PMID- 9868953 TI - Acute liver damage with characteristic apoptotic hepatocytes by ingestion of Aplysia kurodai, a sea hare. AB - A case of acute liver damage by ingestion of Aplysia kurodai, a sea hare is reported. A 40-year-old man, complaining of vomiting and pyrexia after eating a sea hare, was admitted. Laboratory data showed mild liver damage with sustained elevations of aminotransferases. Microscopic findings in the liver biopsy specimen revealed characteristic apoptotic hepatocytes accompanied by mitotic hepatocytes. It is suggested that bioactive substances in the sea hare might induce such apoptosis of hepatocytes in the liver. PMID- 9868954 TI - Aneurysm of the gastroduodenal artery. AB - We report a case of gastroduodenal arterial aneurysm incidentally discovered by abdominal ultrasonogram, in which three-dimensional computed tomography (3D-CT) was useful for diagnosis. An 85-year-old man was hospitalized because of fever and liver dysfunction. Carcinoma of the papilla of Vater was diagnosed by endoscopic retrograde cholangio-pancreatography, while abdominal ultrasonogram and CT scan revealed a round mass measuring 6.2x4.1 cm on the ventral side of the pancreatic head. Three-dimensional CT demonstrated that the mass arose from a branch of the common hepatic artery and projected downward, indicating a gastroduodenal arterial aneurysm. During pancreatoduodenectomy and aneurysmectomy, it was confirmed that this aneurysm arose from the gastroduodenal artery. PMID- 9868956 TI - Cardiac malignant pheochromocytoma with bone metastases. AB - A patient with malignant cardiac pheochromocytoma with bone metastases is described. The primary tumor was located between the pulmonary trunk and the left atrium, while metastatic lesions were found in the iliac bones. Treatments with antihypertensive agents, alpha-methylparatyrosine, and combination chemotherapy with cyclophosphamide, vincristine, and dacarbazine partially improved the patient's symptoms, catecholamine levels, and the metastatic lesion of the iliac bones. However, the primary tumor in the heart progressively increased in size and the patient died of disseminated intravascular coagulation and other various complications about 4 years after the diagnosis of the disease. PMID- 9868955 TI - Long-term patency of an aorta-aortic graft bypass in a patient with Takayasu arteritis. AB - A 39-year-old woman was diagnosed by means of angiography as Takayasu arteritis complicated with severe systemic hypertension due to atypical coarctation of the aorta. Aorta-aortic bypass graft surgery was carried out successfully and hypertension remarkably improved. An evaluation of the graft 23 years later confirmed an almost perfect condition with a very satisfactory clinical status. Extensive long-term follow-up studies have been conducted among young people after surgical repair of aortic coarctation showing encouraging results, however the situation seems to be different for the atypical coarctation in Takayasu patients, since not only the age at the time of intervention affects the outcome, but the different circumstances mainly related to the natural history of the disease. We evaluated the long-term outcome based on similar cases with particular consideration to the extremely rare coexistence of familiar hypercholesterolemia. PMID- 9868957 TI - Partial deficiency of hypoxanthine-guanine phosphoribosyltransferase manifesting as acute renal damage. AB - A 32-year-old man who had had frequent gouty arthritis over the past 17 years, was admitted for acute renal failure. Acute renal failure was improved rapidly after medication was resumed and the patient was sufficiently hydrated. The hypoxanthine-guanine phosphoribosyltransferase (HPRT) activity in the patient had been reduced to about 30% of the normal control. Therefore we considered that this patient suffered from a partial deficiency of HPRT. A point mutation of HPRT gene 68G (guanine) to T (thymine) was detected. This is a mutation that has not been previously reported. Familial analysis indicated that his mother and sister were heterozygotes. PMID- 9868958 TI - Asymptomatic hyponatremia due to inappropriate secretion of antidiuretic hormone as the first sign of a small cell lung cancer in an elderly man. AB - A 72-year-old man was hospitalized with asymptomatic hyponatremia. Despite hyponatremia, urinary sodium excretion with urine osmolality exceeding plasma osmolality persisted. Plasma vasopressin levels were high and independent of plasma osmolality during hypertonic saline infusion. Computed tomography of the chest showed enlarged mediastinal and right hilar lymph nodes. Microscopically, a specimen of lymph nodes obtained by biopsy represented vasopressin-producing small cell lung carcinoma. Chemotherapy plus irradiation improved the hyponatremia. Thus, careful evaluation is necessary to determine the cause of hyponatremia disorders in elderly patients. PMID- 9868959 TI - Mixed medullary-follicular carcinoma and papillary carcinoma of the same thyroid. AB - We report a rare case of mixed medullary-follicular carcinoma and papillary carcinoma of the same thyroid. A 27-year-old Chinese female complained of a single thyroid nodule for 2-3 months. Needle aspiration revealed suspicious papillary carcinoma and thyroidectomy performed later showed mixed medullary follicular carcinoma and papillary carcinoma of the same thyroid which was extremely rare. Whether neoplastic transformation is due to tumorigenic stimulus or just due to the collision phenomenon is still controversial for its etiology. PMID- 9868960 TI - Disturbance of consciousness associated with hypophosphatemia in a chronically alcoholic patient. AB - A 69-year-old man with chronic alcoholism was admitted to our hospital due to disturbance of consciousness and oliguria. Emergency laboratory examination revealed metabolic acidosis, hypoglycemia, hyponatremia, mild liver dysfunction, acute renal failure and rhabdomyolysis. After administration of fluids and nutrients and continuous hemodiafiltration, he recovered from all signs and symptoms except for disturbance of consciousness after 7 days. Since severe hypophosphatemia persisted, we administered adequate phosphates, and then his level of consciousness normalized. We discuss the relationships among alcohol abuse, hypophosphatemia and disturbance of consciousness, and recommend that hypophosphatemia be considered a potential cause of disturbance of consciousness in alcoholic patients. PMID- 9868961 TI - Use of bronchopulmonary lavage for eliminating inhaled fume particles from a patient with arc welder's lung. AB - A 42-year-old man, who had worked as a welder for 20 years, was admitted to our hospital complaining of a dry cough. A chest radiograph showed diffuse small nodular shadows and chest computed tomography revealed small patchy opacities. A transbronchial lung biopsy specimen showed welding fume particles mainly located in alveolar space with mild fibrosis of alveolar septa. In order to prevent further fibrosis, bronchopulmonary lavage (BPL) was performed to eliminate the fume particles. The amount of iron particles derived from the total lavage fluid was 911.7 mg. PMID- 9868962 TI - Benign asbestos pleural effusion associated with pulmonary aspergilloma. AB - A rare case of benign asbestos pleural effusion associated with aspergilloma is reported. A chest radiograph of a 75-year-old Japanese man who was admitted with right chest pain showed a right pleural effusion and nodular shadows in the right apex and left middle lung field. Thoracocentesis revealed an exudate with atypical mesothelial cells. An open lung biopsy showed aspergilloma in the right S2 area and no evidence of malignancy. Many reactive mesothelial cells were found in the pleura. A quantitative asbestos digestion study of the lung tissue biopsy showed high-grade asbestos exposure. PMID- 9868963 TI - Acute pulmonary edema caused by accidental aspiration of sweetened water in two cases of diabetes mellitus. AB - We treated two cases of diabetes mellitus who developed acute pulmonary edema following accidental aspiration of sweetened water for emergency treatment, when they had fallen into hypoglycemic coma following an overdose of injectable insulin. Although they showed hypoxemia and radiological examinations revealed pulmonary edema, they improved by giving only oxygen and antibiotics in a few days. The osmotic pressure of the sweetened water in each case was approximately 2,600 mOsm and 1,900 mOsm. We suppose that the pathogenesis of the pulmonary edema was due to the sweetened water causing water within the pulmonary vessels to permeate into the alveoli. PMID- 9868964 TI - Two cases of myelodysplastic syndrome with extramedullary polyclonal plasma cell proliferation and autoantibody production: possible role of soluble Fas antigen for production of excessive self-reactive B cells. AB - Two cases of myelodysplastic syndrome (MDS) with extramedullary polyclonal plasma cell proliferation and autoantibody production are reported. These cases, which showed leukemic change of refractory anemia with excess of blast (RAEB), developed lymph node swelling and muscle abscess; both were infiltrated mainly with plasma cells, without preceding infection. The proliferation of plasma cells was polyclonal and was proven by negative rearrangement of immunoglobulin heavy chain gene or polyclonal staining of immunoglobulin light chains. These patients showed polyclonal gammopathy and autoantibody production such as positive antinuclear factor and direct antiglobulin test. As was observed in one of the present cases, and as we reported previously, the elevated level of soluble Fas antigen in MDS patients, and its inhibition of apoptotic signaling may be responsible for the excessive accumulation of self-reactive B cells, resulting in these clinical manifestations. PMID- 9868966 TI - Localizing value of bilateral cheiro-oral sensory impairment. AB - We present a case of bilateral cheiro-oral syndrome with paresthesia in the right toes following pontine infarction. The causative lesions were situated in the bilateral paramedian pontine tegmentum (PPM). As the lesion on the left side was larger than that on the right, the foot area in the left medial lemniscus was involved, causing paresthesia in the right toes. Bilateral cheiro-oral sensory impairment is rare, and there have been only 4 previous reports. The responsible lesion of complete bilateral cheiro-oral syndrome showing paresthesia in the hand and mouth region on both sides is in the bilateral PPM, while that of incomplete bilateral cheiro-oral syndrome showing paresthesia around both sides of the mouth and in the unilateral hand is in the PPM contralateral to the hand with paresthesia. Bilateral cheiro-oral sensory impairment can occur only in the brainstem. PMID- 9868965 TI - Atypical rigid form of Huntington's disease: a case with peripheral amyotrophy and congenital defects of a lower limb. AB - We describe a patient showing an atypical phenotype of Huntington's disease (HD), including prominent generalized dystonia, peripheral amyotrophy of the legs with an inverted champagne bottle configuration and pes equinus. The patient also had congenital defects of the lower left leg. Chorea and psychiatric symptoms were not prominent. Polymerase chain reaction assessment revealed 51 CAG repeats in gene IT 15. Magnetic resonance imaging of the brain demonstrated mild atrophy of the pons and cerebellum, and hyperintensity of the transverse pontine fibers and neostriatum on spin-echo images. Peripheral amyotrophy in this case might have resulted from axonal degeneration related to neuronal damage in the central nervous system, although at the present time we cannot confirm it as a new HD phenotype. PMID- 9868967 TI - Efficacy of early plasmapheresis in Bickerstaff's encephalitis. AB - Double filtration plasmapheresis (DFPP) was performed in a patient with Bickerstaff's brainstem encephalitis (BBE) in its early phase. He was a 27-year old male patient suffering from diplopia, facial palsy and drowsiness following upper respiratory tract infection, and had high titers of immunoglobulin G (IgG) antibodies against ganglioside NeuAc(alpha)2-8NeuAc(alpha)2-3Gal(beta)1 3GalNAc(beta)1-4(NeuAc(al pha)2-8NeuAc(alpha)2-3)Gal(beta)1-4Glc(beta)1-1'Cer (GQ1b) in the serum. DFPP effected immediate improvement of his drowsiness, supporting the diagnosis of BBE. Our observations suggest that DFPP during the early phase of BBE efficiently prevents the progression of consciousness disturbances. PMID- 9868968 TI - Adrenal hemorrhage associated with Klebsiella oxytoca bacteremia. AB - Septic adrenal hemorrhage is classically caused by meningococcemia. An autopsied case is presented of a 45-year-old man with adrenal hemorrhage due to Klebsiella oxytoca bacteremia following placement of a central venous catheter. He died 5 hours after developing disseminated intravascular coagulation (DIC). The bacterial entry site may have been the catheter. The cause of death was considered to be pulmonary edema due to bacteremia rather than adrenal insufficiency due to hemorrhage. Septic adrenal hemorrhage should be recognized as a subtype of sepsis rather than adrenal insufficiency, and may be caused in conditions of severe sepsis with DIC, independent of the microorganic variety. PMID- 9868969 TI - Increased plasma lipoprotein(a) level in cardioembolic stroke with non-valvular atrial fibrillation. PMID- 9868970 TI - Spontaneous hypoglycaemia after pancreas transplantation in Type 1 diabetes mellitus. AB - Hypoglycaemia is an important complication of insulin treatment in Type 1 diabetes mellitus (DM). Pancreas transplantation couples glucose sensing and insulin secretion, attaining a distinctive advantage over insulin treatment. We tested whether successful transplantation can avoid hypoglycaemia in Type 1 DM. Combined kidney and pancreas transplanted Type 1 DM who complied with good function criteria (KP-Tx, n = 55), and isolated kidney or liver transplanted non diabetic subjects on the same immunosuppressive regimen (CON-Tx, n = 14), underwent 1-day metabolic profiles in the first 3 years after transplantation, sampling plasma glucose (PG) and pancreatic hormones every 2 hours. KP-Tx had lower PG than CON-Tx in the night and in the morning and higher insulin concentrations throughout the day. KP-Tx had lower PG nadirs than CON-Tx (4.40+/ 0.05 vs 4.96+/-0.16 mmol l(-1), ANOVA p = 0.001). Nine per cent of KP-Tx had hypoglycaemic values (PG < or = 3.0 mmol l(-1)) in the profiles, both postprandial and postabsorptive, whereas none of CON-Tx did (p < 0.02). In conclusion, after pancreas transplantation, mild hypoglycaemia is frequent, although its clinical impact is limited. Compared to insulin treatment in Type 1 DM, pancreas transplantation improves but cannot eliminate hypoglycaemia. PMID- 9868971 TI - Adding metformin versus insulin dose increase in insulin-treated but poorly controlled Type 2 diabetes mellitus: an open-label randomized trial. AB - To compare the effect of adding metformin to insulin therapy with a moderate increase in insulin dose alone in insulin-treated, poorly controlled Type 2 diabetic patients, 47 consecutive such patients (baseline daily dose >0.5 IU kg( 1) and HbA1c >8%) were openly randomized either to a combination of their previous insulin schedule plus metformin (2.55 g daily in three divided doses, n = 24) or to a moderate insulin dose increase (20% of baseline, n = 23). The patient status/biochemical profile was assessed at entry and at 4 months. Among those assigned to insulin + metformin, 18 took the drug. Upon an intention-to treat basis, patients assigned to insulin dose increase had a statistically significant weight gain (1.16+/-1.9 vs 0.3+/-4.5 kg, p < 0.05). Patients assigned to the insulin + metformin regimen experienced a significantly greater fall in HbA1c (-1.87+/-2.16 vs 0.03+/-1.68%, p < 0.01), total cholesterol (-0.56+/-0.89 vs 0.14+/-0.72 mmol l(-1), p < 0.05) and LDL-cholesterol (-0.51+/-0.73 vs 0.19+/ 0.6 mmol l(-1), p < 0.01). These data suggest that adding metformin to insulin in poorly controlled Type 2 DM patients offers an advantage in terms of glycaemic control and lipid plasma profile. PMID- 9868972 TI - Reduced adipocyte insulin sensitivity in Caucasian and Asian subjects with coronary heart disease. AB - The association between insulin resistance and coronary heart disease (CHD) is strong in the British Indian-Asian population. Adipocyte metabolism may contribute to both insulin resistance and CHD. We examined insulin-stimulated glucose uptake in adipocytes and in vivo insulin sensitivity using the fasting insulin resistance index (FIRI) in 60 subjects (45 Caucasian and 15 Asian) with CHD and 30 Caucasian subjects without CHD. In 25 CHD subjects (18 Caucasian and 7 Asian), the relationship between adipocyte insulin sensitivity and non-esterified fatty acid (NEFA) suppression to oral glucose was examined. Compared with controls, the CHD subjects had higher values of fasting insulin [51 (46 to 54) pmol l(-1) vs 36 (31 to 41) pmol l(-1) p< 0.01] and FIRI [1.65 (1.5 to 1.79) vs 1.06 (0.89 to 1.23), p < 0.01]. Among the CHD subjects, the Asians had higher values than Caucasian [insulin 58 (48 to 67) pmol l(-1) vs 48 (44 to 53) pmol l( 1) p < 0.01, FIRI 1.89 (1.44 to 2.13) vs 1.62 (1.4 to 1.79), p< 0.01)]. Insulin stimulated glucose uptake in adipocytes was lower in the CHD than control subjects [56 (50 to 62) vs 115 (75 to 132) attomol min(-1).mm2, p < 0.05], being most reduced among the Asians. It was positively correlated with postprandial NEFA suppression and negatively with insulin release. In conclusion, abnormalities of adipocyte function and insulin sensitivity occur in CHD and may contribute to its aetiology. PMID- 9868973 TI - Long-term glycaemic control directly correlates with glomerular filtration rate in early Type 1 diabetes mellitus before the onset of microalbuminuria. AB - Hyperfiltration occurs early in diabetes mellitus and has been implicated in the development of microalbuminuria. Our aim was to re-examine the controversial relationship between glycaemic control and glomerular filtration (GFR) in normoalbuminuric, normotensive, non-obese patients with short duration Type 1 diabetes mellitus (DM). We studied 75 Type 1 DM patients, 35 male, aged 18-42 years, with a duration of diabetes of 4-8 years. GFR was determined by inulin clearance; hyperfiltration was defined as above 145 ml min(-1) 1.73 m(-2) (equivalent to 2 SD above mean for a control population). Analysis was by paired Student's t-testing and linear regression. GFR correlated significantly with HbA1c (r= 0.47, p < 0.0001) and fructosamine (r= 0.24, p = 0.035). Mean HbA1c and fructosamine in the 13 patients with hyperfiltration was significantly higher than in the rest of the group (HbA1c: 9.2% (95% C.I. 7.9-10.4%) vs 7.6 % (7.2 7.9), p= 0.002; fructosamine: 479 micromol l(-1) (450-507) vs 410 micromol l(-1) (388-432), p = 0.009. This significant difference persisted even when the two highest values of HbA1c or fructosamine were removed from analysis. Effective renal plasma flow, assessed by PAH clearance, also correlated in all patients with HbA1c (r=0.31, p=0.039). We conclude that poor glycaemic control directly correlates with hyperfiltration and renal hyperperfusion in early Type 1 DM. PMID- 9868974 TI - Evidence of an increasing prevalence of diagnosed diabetes mellitus in the Poole area from 1983 to 1996. AB - This study examined the prevalence of diagnosed diabetes mellitus in a defined population over 13 years by undertaking cross-sectional surveys on 3 occasions between 1983 and 1996. The study population consisted of all the people registered with 10 general (primary care) practices at the time of each survey; 90660 in 1983/4; 97122 in 1988/9; and 86287 in 1996. Ascertainment of cases was by a surveillance programme in general practice and the hospital diabetes department. The number of diabetic patients increased significantly over the study period: in 1983/4, there were 917 patients, crude prevalence 1.01% (95% CI 0.95-1.08%); in 1988/9, 1150 patients, crude prevalence 1.17% (1.12-1.25%); and in 1996, 1604 patients, crude prevalence 1.86% (1.77-1.95%). The prevalence adjusted to the age and sex distribution of the UK was 0.97% (95% CI 0.90-1.03%) in 1983/4, 1.05% (0.99-1.11%) in 1988/9 and 1.55% (1.48-1.63%) in 1996. The main increase in prevalence was due to Type 2 diabetes mellitus, crude prevalence 0.75% (95% CI 0.69-0.81%) in 1983/4, 0.92% (0.86-0.98%) in 1988/9 and 1.52% (1.44 1.60%) in 1996 rather than Type 1 diabetes mellitus, crude prevalence 0.25% (0.21 0.28%) in 1983/4, 0.25% (0.22-0.28%) in 1988/9 and 0.34% (0.30-0.38%) in 1996. During the study period, the crude prevalence of diagnosed diabetes was significantly greater in men than women; in 1983/4 men 1.1% (95% CI 1.00-1.20%) versus women 0.93% (0.84-1.02%); in 1988/9, men 1.31% (1.21-1.41%) versus women 1.07% (0.98-1.16%); and in 1996, men 2.13% (2.00-2.27%) versus women 1.60% (1.49 1.72%). This difference was statistically significant in the 1988/9 and 1996 surveys. In conclusion, over 13 years there was a significant increase of 83.6% in the prevalence of diagnosed diabetes mellitus in the Poole area, with the UK age and sex adjusted prevalence increasing by 60.7%. PMID- 9868976 TI - Withholding thrombolysis in patients with diabetes mellitus and acute myocardial infarction. AB - The benefits of thrombolytic therapy in a patient with diabetes having a myocardial infarction are now well accepted but this treatment may be withheld inappropriately because of concerns about retinal haemorrhage. We therefore examined whether junior doctors alter their use of thrombolysis for the treatment of acute myocardial infarctions according to the type of diabetic retinopathy present. A questionnaire asking whether thrombolysis would be given to a 50-year old male smoker with insulin-treated diabetes and an acute anterior MI was shown, with four unlabelled retinal photographs, to all doctors prescribing thrombolytic therapy in a south London teaching hospital and an affiliated district general hospital. In all, 24 medical SHOs, 16 medical registrars/specialist registrars, 3 medical senior registrars, and 23 casualty SHOs were interviewed. Of these 89% would thrombolyse such a patient with normal fundi, 55% with background diabetic retinopathy, 54 % if this also involved the macula, and 26% if they saw proliferative retinopathy. The more senior grades were more aggressive in their approach. As we believe that all patients with an acute anterior myocardial infarction and diabetes should be considered for thrombolysis irrespective of their retinal appearance these results suggest thrombolytic therapy is being withheld inappropriately. PMID- 9868975 TI - Rapid gastric emptying of a liquid meal in long-term Type 2 diabetes mellitus. AB - Both delayed and accelerated gastric emptying rate (GER) have been reported in patients with diabetes mellitus. Delayed GER has been attributed to autonomic neuropathy in established diabetes but rapid GER was demonstrated in early Type 2 diabetes. The aim of the study was to investigate rapid gastric emptying in a group of people with long-duration Type 2 diabetes. GER of a radiolabelled liquid meal was studied scintigraphically in 20 Type 2 patients with a mean (+/-SEM) duration of diabetes 13 (+/-1) years. The 50% emptying time (t50) for the liquid meal was shorter in diabetic patients (29.6+/-2.1 min) than in controls (39.2+/ 1.9 min; p<0.0005). Accelerated emptying (t50 value below the shortest t50 of controls) was evidenced in 14/20 patients and delayed emptying (t50 value exceeding the upper t50 of controls) in none. Patients with accelerated GER were comparable for BMI, diabetes duration, HbA1c and fasting glycaemia to those with normal GER. Rapid GER for liquids was found in the presence or absence of autonomic neuropathy. Seven of the patients with rapid emptying of the liquid meal were reassessed using a solid meal. Only one patient demonstrated rapid emptying of the solid meal, which was normal in 3 and delayed in 3 patients. In conclusion, accelerated GER can be found in long-term Type 2 diabetes but there is no concordance between GER of a liquid and a solid meal. PMID- 9868977 TI - Apolipoprotein E genotype is a determinant of low-density lipoprotein cholesterol and of its response to a low-cholesterol diet in Type 1 diabetic patients with elevated urinary albumin excretion. AB - The effect of the apolipoprotein (apo) E genotype on the lipoprotein response to a 1 year low cholesterol diet (200 mg cholesterol per day) was evaluated in 36 patients with Type 1 diabetes mellitus with albuminuria between 10 and 200 microg min(-1). Apo E genotype was characterized by polymerase chain reaction and restriction isotyping. In 11 IDDM patients with at least one epsilon4 allele (apo E4 group), baseline serum total and low density lipoprotein (LDL) cholesterol were higher (p < 0.05 for both) than in 25 patients without an epsilon4 allele and with at least one epsilon3 allele (apo E3 group). Dietary counselling resulted in a similar decrease in cholesterol intake in both groups, whereas linoleic acid did not change. In the apo E4 group, serum total and LDL cholesterol at follow-up fell (p < 0.01 for both) to levels that were not different from those in the apo E3 group, and the changes in these parameters were greater (p < 0.02) than those in the apo E3 group. We conclude that the apo E4 allele is associated with atherogenic lipoprotein abnormalities in Type 1 DM patients with minor elevations in albuminuria when they use their habitual diet. Apo E4 carrying patients respond better to a low cholesterol diet. PMID- 9868978 TI - Gestational diabetes mellitus in teenage pregnancy: a case-control study. AB - In a retrospective study, teenage Asian pregnancies with gestational diabetes managed over a 4-year period were compared with a group of age and parity matched controls (2 for each study case) to determine the incidence of gestational diabetes and its impact on the pregnancy outcome. The incidence of gestational diabetes in teenage pregnancy was 5.4% (33/611), and accounted for 1.4% of all the cases of gestational diabetes. There was no difference in the maternal anthropometric parameters or antenatal complications, but the study group had a higher incidence of postpartum haemorrhage (p = 0.010), greater amount of estimated blood loss at delivery (p = 0.016), a trend towards a higher incidence of large-for-gestational age infants, a higher incidence of admission to the neonatal unit (p = 0.024), mostly due to meconium-stained liquor for observation (p = 0.014), and a lower first minute Apgar score (p = 0.012). Our findings support the recommendation that in ethnic groups with a high prevalence of diabetes, universal as opposed to age-limited screening for gestational diabetes should be undertaken. PMID- 9868979 TI - No deterioration in insulin sensitivity, but impairment of both pancreatic beta cell function and glucose sensitivity, in Japanese women with former gestational diabetes mellitus. AB - To identify the primary pathogenic factors involved in the development of Type 2 diabetes mellitus (DM), we studied Japanese women with former gestational diabetes mellitus (GDM) who are at risk for the later development of Type 2 DM. We used the minimal model analysis derived from frequently sampled intravenous glucose tolerance test (FSIGT). The subjects consisted of eight non-obese women with a history of GDM and eight non-obese normal women as control subjects. The 75 g oral glucose tolerance test (75 g OGTT) performed within 6 months of delivery confirmed that all the subjects with former GDM had a normal glucose tolerance. Insulin sensitivity (SI) derived from the minimal model analysis was not different between the two groups. Glucose effectiveness at zero insulin (GEZI), reflecting tissue glucose sensitivity, was significantly lower in former GDM patients than in control subjects (1.18+/-0.34 vs 2.26+/-0.29 x 10(-2) min( 1), p < 0.05). The early phase insulin secretion found in FSIGT was markedly reduced to 56% of that observed in control subjects (1250+/-87.4 vs 2223+/-304.3 pmol l(-1) min, p < 0.01). Our results indicate that in former GDM patients, who are japanese and non-obese, impairment of the acute insulin response to glucose and a decrease in tissue glucose sensitivity rather than insulin sensitivity are the primary pathogenic factors involved. PMID- 9868980 TI - Diabetes mellitus in Egypt: glycaemic control and microvascular and neuropathic complications. AB - We performed a cross-sectional, population-based survey of persons 20 years of age and older living in Cairo and surrounding rural villages. The purpose was to describe glycaemic control and the prevalence of microvascular and neuropathic complications among Egyptians with diagnosed diabetes, previously undiagnosed diabetes, impaired glucose tolerance, and normal glucose tolerance. A total of 6052 households were surveyed. The response rate was 76% for the household survey and 72% for the medical examination. Among people with previously diagnosed diabetes, mean haemoglobin A1c, was 9.0%. Forty-two per cent had retinopathy, 21% albuminuria, and 22% neuropathy. Legal blindness was prevalent (5%) but clinical nephropathy (7%) and foot ulcers (1%) were uncommon in persons with diagnosed diabetes. Among people with diagnosed diabetes, microvascular and neuropathic complications were associated with hyperglycaemia. Retinopathy was also associated with duration of diabetes; albuminuria with hypertension and hypercholesterolaemia; and neuropathy with age, female sex, and hypercholesterolaemia. Albuminuria was as common in people with previously undiagnosed diabetes (22%) as those with diagnosed disease (21%). Mean haemoglobin A1c was lower (7.8%) and retinopathy (16%) and neuropathy (14%) were less prevalent in people with previously undiagnosed disease. Ocular conditions, blindness, and neuropathy were prevalent in the non-diabetic population. The microvascular and neuropathic complications of diabetes are a major clinical and public health problem in Egypt. PMID- 9868981 TI - Glucose intolerance and associated factors in the Fergana Valley, Uzbekistan. AB - Prevalence of glucose intolerance and other noncommunicable diseases has been examined in subjects aged 35 years and over in semirural and urban communities in the Fergana Valley in the eastern part of Uzbekistan, Central Asia. Diabetes and impaired glucose tolerance (IGT) were diagnosed according to the recommendations of the latest WHO Study Group on diabetes. Crude prevalence of diabetes was 9% and 5%, respectively, in semirural men and women, 13% and 9% in urban men and women. Crude prevalence of impaired glucose tolerance (IGT) was 6% and 9%, respectively, in semirural men and women, 9% and 8% in urban men and women. After adjustment for non-response, prevalence of diabetes was 5% and 4%, respectively, in semirural men and women and 8% in both urban men and women. Adjusted prevalence of IGT was 4% and 8%, respectively, in semirural men and women, 5% and 6% in urban men and women. The majority of subjects with a prior diagnosis of diabetes were being treated with oral hypoglycaemic agents. Almost one-half of subjects in both communities had body mass index of 25 kg m(-2) or greater. Central obesity (waist-hip ratio 0.95 or greater for men, 0.85 or greater for women) was observed in over one-quarter of subjects in both communities. Clinical hypertension was not frequent by international standards (9% in semirural subjects and 13% in urban subjects) but a number of subjects who were clinically normotensive claimed to be taking antihypertensive medication. It is concluded that glucose intolerance and central obesity are common in this region of Uzbekistan, about which there was previously little information. PMID- 9868982 TI - Resolution of lipohypertrophy following change of short-acting insulin to insulin lispro (Humalog). AB - Lipohypertrophy as a local complication of insulin therapy is well recognized. Despite improvements in insulin purity and the introduction of recombinant human insulin its prevalence has remained high. Rotation of injection sites can reduce the frequency of the problem but does not abolish it. The importance of this complication is not only cosmetic but also in its impact on insulin absorption, and hence glycaemic control. We report a patient who had intractable lipohypertrophy with human recombinant insulin but experienced no such problem when converted onto the insulin analogue lispro. We suggest that the faster speed of absorption of insulin lispro may lead to less hypertrophic stimulation of subcutaneous adipocytes. This difference may be clinically useful in susceptible individuals. PMID- 9868983 TI - A case of diabetic muscle infarction in Japan. AB - Diabetic muscle infarction (DMI) is a rare complication of diabetes mellitus. We report the first recorded case in Japan. A 45-year-old Japanese woman presented with severe pain in the left antero-medial thigh. She had a 14-year history of Type 2 diabetes mellitus (DM). She had first noticed pain in her left thigh after a walk 2 weeks prior to presentation. The pain worsened progressively. She noticed a firm mass in her left thigh. T2-weighted magnetic resonance imaging (MRI) demonstrated a high-intensity signal in the muscle bulk of the anterior component of the left thigh. A needle biopsy of the mass showed necrosis. She was treated with bedrest and an antiplatelet agent. The mass disappeared 8 weeks after admission. DMI is a rare complication of poorly controlled diabetes mellitus. Twenty-seven cases with DMI have been reported in the English literature but we believe this is the first Japanese case with DMI. PMID- 9868984 TI - Type 2 diabetes mellitus: the grand overview. AB - Type 2 diabetes currently accounts for over 100 billion dollars in annual healthcare expenditure in the United States and 28% of the national (Medicare) healthcare budget for elderly Americans. In our inner-city hospital, 20% of all 950 beds are occupied by patients with diabetes; and 28-38% of patients receiving cardiac care in Coronary Care Units, catheterization laboratories or cardiovascular surgery, have diabetes as an underlying disorder. Both computer modelling and controlled clinical trials suggest that intensive therapy of diabetes can reduce significantly the morbidity and costs associated with this increasingly common disorder. Early detection of carbohydrate intolerance holds great promise for preventing the onset, progression and complications of Type 2 diabetes. To date our efforts have been futile, with 20% of newly diagnosed Type 2 diabetic patients already complicated by retinopathy and 14% complicated by peripheral vascular disease. It is now clear that high-risk individuals can be identified, and intervention trials are underway to test the hypothesis that Type 2 diabetes (and its attendant cardiovascular risks) can be prevented. The Study to Prevent Non-Insulin-Dependent Diabetes Mellitus (STOP NIDDM) in Canada and Europe has randomized 1200 individuals with impaired glucose tolerance (IGT) into a three-year trial to prevent disease progression. The Diabetes Prevention Program (DPP) in the US has randomized almost 3000 individuals with IGT into a six-year, three-arm study testing the efficacy of intensive lifestyle and pharmacological therapy in disease progression. Together, these studies should provide a public health model for the recognition of high-risk individuals and interventions to stem the epidemic of Type 2 diabetes. For those patients suffering with Type 2 diabetes already, pancreas transplantation remains an extreme intervention with the potential for 'curing' diabetes. Although applied usually to patients with Type 1 diabetes, experience is accumulating of transplantation in Type 2 diabetic patients with end-stage renal disease. Outcomes for these individuals are as good as for Type 1 diabetes. Islet-cell transplants, in fact, have been more successful in Type 2 diabetes compared with Type 1. Improved islet-cell availability, better immunosuppression, and the possibility of antigen masking make this technology a major hope for the future. PMID- 9868985 TI - Managing diabetes: lessons from Type 1 diabetes mellitus. AB - Glycaemic control is a key component of the successful management of Type 1 and Type 2 diabetes mellitus. Hypoglycaemia is the limiting factor in the management of diabetes because current glucose-lowering regimens are imperfect, defences against decreasing glucose levels in Type 1 and probably Type 2 diabetes are compromised, and low glucose levels have a devastating effect on the brain. Usually, hypoglycaemia precludes the maintenance of normal glucose levels. However, attempts to circumvent the barrier of hypoglycaemia safely are worthwhile because shifting glucose levels towards the non-diabetic range reduces the long-term complications of diabetes. Patient education and empowerment, appropriate self-monitoring of blood glucose, flexible drug regimens, individualized and prudent glycaemic goals, and ongoing professional support are fundamental. Iatrogenic hypoglycaemia is the result of the interplay between excess insulin and compromised glucose counter-regulation in Type 1 and probably Type 2 diabetes. Conventional and newly recognized risk factors must be addressed. Relative or absolute excess insulin occurs when: insulin (or insulin secretagogue) doses are excessive, ill-timed or of the wrong type; the influx of exogenous glucose, endogenous glucose production or insulin clearance are decreased; and insulin-independent glucose utilization or insulin sensitivity are increased. The drug regimen, food ingestion, exercise and alcohol use are under the direct control of the patient and the healthcare provider, and regimen adjustments can be used to address insulin sensitivity and clearance. Unfortunately, these conventional risk factors explain only a minority of episodes of severe hypoglycaemia and therefore the issue of compromised glucose counter-regulation must also be addressed. It is imperative to investigate the patient history for hypoglycaemia unawareness because short-term (e.g. 2 weeks) scrupulous avoidance of hypoglycaemia can restore awareness and improve defective glucose counter-regulation. Until methods of perfect insulin replacement or release are developed, improved regimens and pharmacological methods to minimize hypoglycaemia particularly during the night can be used safely to improve overall glycaemic control. PMID- 9868986 TI - Improving glycaemic control with current therapies. AB - A considerable proportion of Type 2 diabetic patients suffer from hyperglycaemic symptoms and therefore experience reduced quality of life. Furthermore, increasing evidence suggests that poor glycaemic control is associated with a risk that late complications will develop. The traditional stepped approach to therapy often results in a reluctance to escalate therapy to keep up with the progression of the disease, and therefore new strategies are needed to improve the results. Type 2 diabetes is a heterogeneous disorder, and hyperglycaemia is the result of deficient insulin secretion and insulin resistance; and the natural course of the disease is progression of hyperglycaemia. The therapy should be tailored to match the different needs of individual patients. Diet and exercise are essential to support all other therapies, but are often overlooked and may not be effective alone. The effectiveness of oral hypoglycaemic agents (OHAs) depends on the patients having sufficient insulin secretory capacity. These agents are therefore of little benefit to patients with profound beta-cell failure. The combination of oral agents from two different pharmaceutical groups can be more effective than monotherapy, but in many patients insulin deficiency ensues and hyperglycaemia progresses. In principle, insulin therapy should always be able to lower glucose levels; improved glycaemic control is achieved in most patients, followed by amelioration of hyperglycaemic symptoms and improvements in quality of life. However, near-normoglycaemia may be difficult to achieve with the pharmacological limitations imposed by the preparations available, the methods of administration, and the ability and motivation of the patients. Importantly, insulin therapy should be tailored to meet the individual needs of the patients, and patients should be taught self-adjustment of doses based on self-monitoring of blood glucose levels. A considerable proportion of Type 2 diabetic patients (primarily the young and lean) require multiple-dose regimens. Combination therapy with OHAs and insulin might offer an advantage to some patients, and a recent study from Finland suggests that the combination of bedtime insulin and daytime metformin may be superior to other bedtime insulin regimens. There is still some way to go to devise an optimal therapy for Type 2 diabetes. PMID- 9868987 TI - What works? Success stories in Type 2 diabetes mellitus. AB - The increasing prevalence of Type 2 diabetes mellitus has made diabetes a major public health problem worldwide. Healthcare systems are faced with the major challenge of developing innovative approaches to improving the prevention of diabetes and associated complications, particularly in populations undergoing rapid cultural development. Examples of community-based primary prevention projects in two Native communities in Canada that have high incidences of diabetes are reviewed. These projects are notable for the active participation of the communities, the collaborative relationship that was established between the communities and the researchers, and the way that traditional native culture and beliefs were incorporated into the intervention design. This phenomenon has impacted on the success of the projects and has enhanced their long-term sustainability. PMID- 9868988 TI - What works? The Diabetes Care and Information Center. AB - This report focuses on the contribution of a referral practice that offers specialist diabetes care to a multi-ethnic population in New York, and describes the process and outcome of an intensive management policy that is particularly strong on communication. The Diabetes Care and Information Center (DCIC), which practices shared care with local primary care physicians, has developed a programme of intensive management for Type 1 and Type 2 diabetes based on a detailed initial patient assessment by a physician, nurse-educator and dietician team. The assessment is followed by an individualized programme of patient education that includes a combination of individual and group education, and counselling. Patients are followed up primarily by an educator or a physician, with frequent interdisciplinary communication to optimize self-care procedures. Medication programmes for patients with Type 2 diabetes include monotherapy and combination therapy with oral agents and single or multiple doses of insulin. Metabolic improvement has been demonstrated by significant changes in mean HbA1c. In addition to initial assessment and routine metabolic follow up, the DCIC provides routine screening and treatment for micro- and macrovascular complications of diabetes, which results in early referral to podiatrists, vascular surgeons and ophthalmologists for appropriate preventive care. PMID- 9868989 TI - Repaglinide--prandial glucose regulator: a new class of oral antidiabetic drugs. AB - The highest demand on insulin secretion occurs in connection with meals. In normal people, following a meal, the insulin secretion increases rapidly, reaching peak concentration in the blood within an hour. The mealtime insulin response in patients with Type 2 diabetes is blunted and delayed, whereas basal levels often remain within the normal range (albeit at elevated fasting glucose levels). Restoration of the insulin secretion pattern at mealtimes (prandial phase)--without stimulating insulin secretion in the 'postabsorptive' phase--is the rationale for the development of 'prandial glucose regulators', drugs that are characterized by a very rapid onset and short duration of action in stimulating insulin secretion. Repaglinide, a carbamoylmethyl benzoic acid (CMBA) derivative is the first such compound, which recently has become available for clinical use. Repaglinide is very rapidly absorbed (t(max) less than 1 hour) with a t1/2 of less than one hour. Furthermore, repaglinide is inactivated in the liver and more than 90% excreted via the bile. The implications of tailoring repaglinide treatment to meals were examined in a study where repaglinide was dosed either morning and evening, or with each main meal (i.e. breakfast, lunch, dinner), with the total daily dose of repaglinide being identical. The mealtime dosing caused a significant improvement in both fasting and 24-hour glucose profiles, as well as a significant decrease in HbA1c. In other studies, repaglinide caused a decrease of 5.8 mmol x l(-1) in peak postprandial glucose levels, and a decrease of 3.1 mmol x l(-1) in fasting levels with a reduction in HbA1c of 1.8% compared with placebo. In comparative studies with either sulphonylurea or metformin, repaglinide caused similar or improved control (i.e. HbA1c, mean glucose levels) and the drug was well tolerated (e.g. reported gastrointestinal side-effects were more than halved when patients were switched from metformin to repaglinide). A hallmark of repaglinide treatment is that this medication follows the eating pattern, and not vice versa. Hence the risk of developing severe hypoglycaemia (BG < or = 2.5 mmol x l(-1)) in connection with flexible lifestyles should be reduced. This concept was examined in a study in which patients well controlled on repaglinide skipped their lunch on one occasion. When a meal (i.e. lunch) was skipped--so was the repaglinide dose, whereas in the comparative group on glibenclamide the recommended morning and evening doses were taken. Twenty-four per cent of the patients in the glibenclamide group developed severe hypoglycaemia, whereas no hypoglycaemic events occurred in the group receiving repaglinide. However, in long-term studies the overall prevalence of hypoglycaemia was similar to that found with other insulin secretagogues. In summary, current evidence shows that the concept of prandial glucose regulation offers good long-term glycaemic control combined with a low risk of severe hypoglycaemia with missed meals. The concept should meet the needs of Type 2 diabetic patients, allowing flexibility in their lifestyle. PMID- 9868990 TI - Breaking the barriers for improved glycaemic control: primary care and secondary care interface. AB - The prevalence of diabetes is predicted to increase dramatically over the next 30 years, mostly as a result of Type 2 diabetes. To manage this increased burden, the following measures must be implemented: diabetes care must be delivered in the most suitable locations; people with diabetes must remain in contact with the appropriate healthcare professionals; and health professionals must use the most effective methods to treat their patients. The structure for diabetes care suggested by the World Health Organization uses terminology that is familiar universally. The reality, in terms of what can be delivered at primary- and secondary-care level will, however, differ greatly according to the extent of service development and the resources available. The optimum forum for diabetes care should have the advantages of the primary-care setting and, when required, the main advantages of the secondary-care setting, i.e. specialized knowledge and more sophisticated facilities. In most healthcare systems, particularly those without financial disincentives to the patient to be followed up in hospital, the tendency remains for secondary care to 'capture' and retain patients for longer than is required clinically. The end results are the overloading of hospital services, inappropriate use of resources and the tendency for patients to default from care because of the relative inaccessibility and remoteness of the hospital service. Issues with special reference to different models of diabetes care, use of patient registers and clinical guidelines, and the perceptions of patients regarding the successful use of services are presented. PMID- 9868991 TI - The elderly Type 2 diabetic patient: special considerations. AB - Diabetes mellitus is a common disease in older people, with almost 50% of Type 2 diabetic patients being over 60 years of age; despite this, half of older people with frank diabetes are not diagnosed. While insulin resistance is common in older people, large numbers also have impaired insulin secretion. Age, body habitus and physical activity all play a role in the pathogenesis of hyperglycaemia associated with diabetes mellitus. Leptin levels relate to insulin resistance in older people and amylin secretion is associated with delayed return of glucose levels to baseline. Depression, impaired cognitive function, and lack of recognition of thirst and subsequent dehydration are important factors to be taken into account in the management of older diabetic patients, who may also have impaired physical function, an increased rate of injurious falls, and increased prevalence of pressure ulcers, amputations and tuberculosis. Hyperglycaemia can result in a decreased pain threshold and incontinence. Dietary management plays less of a role in older diabetic patients but exercise, with a particular emphasis on balance and stability, is an important component of the management and treatment of older diabetic patients. The use of metformin as a treatment should be avoided in patients over 80 years of age because of declining kidney function. Insulin therapy is an option but as hypoglycaemia is related to advancing age, patients should be monitored carefully for the development of hypoglycaemia. Care providers also play an important role in the management of older people with diabetes mellitus. Glycaemic control can be obtained with minimal side-effects in most older diabetics including those patients in nursing homes. PMID- 9868992 TI - Lowering the risk of visual impairment and blindness. AB - Diabetic retinopathy remains the leading cause of visual disability and blindness among professionally active adults in economically developed societies, which is of particular concern because the prevalence and incidence of Type 2 diabetes mellitus is expected to increase sharply during the next decade. Retinopathy is fundamentally similar in Type 1 and Type 2 diabetes mellitus, and it is widely accepted that if detected and treated early, loss of vision and blindness from diabetic retinopathy may be prevented. Studies such as the Diabetes Control and Complications Trial (DCCT), the United Kingdom Prospective Diabetes Study Group (UKPDS), the Diabetic Retinopathy Study (DRS) and the Early Treatment Diabetic Retinopathy Study (ETDRS) have established accepted guidelines for the management of diabetic retinopathy. Good metabolic control is particularly important in the early phases of the disease, and will delay the onset of retinopathy and decrease the rate of progression. When advanced stages of retinopathy are reached, laser photocoagulation is effective in decreasing the development to blindness by over 50%. However, preventable blindness still occurs despite the tight control of blood glucose levels and the use of retinal photocoagulation. To reduce the risk of visual impairment and blindness caused by diabetes, diabetic patients must be taught how to control their blood glucose levels, regular eye examinations must be carried out and the conditions for timely laser photocoagulation must be created. The implementation of screening and treatment programmes for visual impairment in diabetes has proved to be worthwhile in terms of costs and health benefits. PMID- 9868993 TI - Preventing end-stage renal disease. AB - Interest in evidence-based medicine is increasing greatly, with the focus on treatment that prevents organ failure and that may prolong life. Type 1 and Type 2 diabetes are conditions associated with increased mortality, mainly as a result of renal and cardiovascular diseases, and blindness. All three complications usually occur together. In recent years, more focus has been placed on treating patients early to prevent future organ damage. Microalbuminuria is an important intermediary end-point that correlates strongly with future advanced renal disease, retinopathy and mortality. Several trials have studied patients with microalbuminuria and also patients in more advanced stages of the disease who have proteinuria (termed overt nephropathy). Recent evidence indicates that achieving optimal glycaemic control reduces the risk of an increase in urinary albumin excretion before the development of microalbuminuria. Angiotensin converting enzyme (ACE) inhibitors are effective in reducing microalbuminuria, partly independent of their blood pressure reducing effects. In Type 1 and Type 2 diabetic patients with microalbuminuria, long-term treatment with ACE inhibitors (7-8 years) prevents the predicted decrease in glomerular filtration rate (GFR); optimal glycaemic control is also important in preventing the decline in GFR. This is important because GFR is usually well preserved in Type 1 and Type 2 diabetic patients with microalbuminuria and a predicted decline in GFR can therefore be prevented. In overt renal disease, studies that focused mostly on Type 1 diabetic patients have shown that the rate of decline in GFR can be reduced. Long-term studies in Type 1 diabetic patients have also demonstrated that mortality caused by end-stage renal disease can be postponed. Mortality associated with cardiovascular diseases, e.g. myocardial infarction, is reduced more effectively in diabetic patients treated with ACE inhibitors and beta blockers than in non-diabetic patients treated with the same drugs. Screening for microalbuminuria, the attainment of optimal glycaemic control, and early treatment with ACE inhibitors and other antihypertensive drugs are necessary to prevent progression of diabetic complications, especially diabetic nephropathy. However, there is some controversy about the initial use of calcium channel blockers. In conclusion, early achievement of improved glycaemic control is the most important factor in the prevention of diabetic complications. Antihypertensive treatment is clearly also important. PMID- 9868994 TI - Lowering the risk of neuropathy, foot ulcers and amputations. AB - Peripheral neuropathy is one of the most common long-term complications of Type 2 diabetes. A population-based study in the north of England showed that 42% of Type 2 diabetic patients had clinical evidence of neuropathy. The Diabetes Control and Complications Trial (DCCT) has shown that the incidence of neuropathy in Type 1 diabetes can be reduced by over 50% with intensive therapy and optimal glycaemic control. Hyperglycaemia is believed to be a major aetiological factor in the development of neuropathy in Type 2 diabetes. Neuropathy cannot be diagnosed through history alone; therefore, careful examination of the feet for evidence of sensory loss and an assessment of the circulation must form part of the annual review of each patient. Peripheral somatic and autonomic neuropathy, together with peripheral vascular disease, are major contributing factors to the development of foot ulcers. In addition, abnormalities of foot shape (e.g. claw toes, prominent metatarsal heads) and the presence of plantar callus are signs of foot-ulcer risk. Effective patient education can reduce the incidence of foot ulceration and amputation by over 50%; therefore, all patients with a high risk of foot ulcers should be informed and, if indicated, referred for regular podiatry. The team approach to diabetic foot problems is an effective method of providing treatment for active ulcers. This should be followed by appropriate education, the provision of follow up and if indicated, suitable footwear and hosiery. Key members of the team are the podiatrist, the specialist nurse and the orthotist; medical staff may include the diabetologist and a vascular or orthopaedic surgeon. Thus, the risk of foot ulceration and amputation can be reduced by careful screening and patient education, without the need for expensive equipment. PMID- 9868995 TI - Accelerated complications in Type 2 diabetes mellitus: the need for greater awareness and earlier detection. AB - Persistent hyperglycaemia is the underlying pathogenic factor responsible for chronic diabetic complications in Type 1 and Type 2 diabetes mellitus. In Type 1 diabetes, diagnosis is made soon after the onset of hyperglycaemia and several years are required for the resultant complications to appear clinically. The onset of Type 2 diabetes is insidious and is usually recognized only 5-12 years after hyperglycaemia develops. During this period of undiagnosed diabetes, hyperglycaemia, in combination with lifestyle factors (physical inactivity, alcohol use, smoking), and other metabolic (dyslipidaemia, obesity, insulin resistance) and haemodynamic (hypertension) abnormalities frequently associated with Type 2 diabetes, promote the initiation and progression of micro- and macrovascular complications. Furthermore, when blood glucose levels are increased only slightly and no symptoms are apparent, the physician may be reluctant to diagnose Type 2 diabetes or start treatment. This delay in diagnosing the disease results in a high prevalence of chronic complications at the time of actual diagnosis. Indeed, when Type 2 diabetes is diagnosed, cardiovascular disease and neuropathy are found in approximately 10% of cases, and retinopathy and nephropathy in 15-20%. All healthcare providers should be aware of this phenomenon, which may be termed 'accelerated complications', and should plan thorough screening programmes for these conditions at diagnosis. All reversible risk factors associated with diabetes should be identified and treated. When acute metabolic derangements and infections are not the main causes of morbidity and mortality in diabetes, the costs of diabetes care are related mainly to chronic complications of the disease. Therefore, because of the high frequency of Type 2 diabetes, the most efficient method of reducing costs is to increase awareness and secure earlier detection that leads to fast and aggressive treatment of the accelerated chronic complications often seen in Type 2 diabetes. PMID- 9868996 TI - The postprandial state and risk of cardiovascular disease. AB - Metabolism in man is regulated by complex hormonal signals and substrate interactions, and for many years the clinical focus has centred on the metabolic and hormonal picture after an overnight fast. More recently, the postprandial state, i.e. 'the period that comprises and follows a meal', has received more attention. The oral glucose tolerance test (OGTT), although highly non physiological, has been used largely as a model of the postprandial state. Epidemiological studies have shown that, when 'impaired', oral glucose tolerance is associated with an increased risk of cardiovascular disease. Postprandial hyperlipidaemia has been investigated more recently in epidemiological, mechanistical and intervention studies, most of which indicate that high postprandial triglyceride levels, and particularly postprandial rich triglyceride remnants, constitute an increased risk for cardiovascular disease. Recent studies have shown that excessive postprandial glucose excursions are accompanied by oxidative stress and, less well known, activation of blood coagulation (increase in circulating D-dimers and prothrombin fragments). The mechanisms through which increased postprandial glucose levels and lipid concentrations may damage endothelial cells on blood vessel walls appear to be complex. These mechanisms include the activation of protein kinase C, increased expression of adhesion molecules, increased adhesion and uptake of leucocytes, increased production of proliferative substances such as endothelin, increased proliferation of endothelial cells, increased synthesis of collagen IV and fibronectin, and decreased production of nitric oxide (NO). In conclusion, the 'postprandial state' cumulatively covers almost half of the nycthemeral period, and its physiology involves numerous finely regulated motor, secretory, hormonal and metabolic events. Epidemiological and mechanistical studies have suggested that perturbations of the postprandial state are involved in cardiovascular disease. Correcting the abnormalities of the postprandial state must form part of the strategy for the prevention and management of cardiovascular diseases, particularly those that are associated with diabetes mellitus. PMID- 9868997 TI - Reducing the cardiovascular consequences of diabetes mellitus. AB - Type 2 diabetes is an important, independent risk factor for cardiovascular disease. As a result of the pandemic explosion of Type 2 diabetes, reducing the human and financial consequences of the disease is a major healthcare concern. In primary prevention, antihypertensive treatment, aspirin and lipid-lowering intervention are effective at reducing the incidence of coronary heart disease although the beneficial effect of optimal glycaemic control remains to be conclusively proven. Type 2 diabetic patients with acute myocardial infarction should receive a specific programme of intensive treatment, which includes optimal glycaemic control using intensive insulin treatment, thrombolysis, and early use of beta-blockers and angiotensin-converting enzyme inhibitors. Secondary prevention is also essential because beta-blockers and lipid interventions are more effective in Type 2 diabetic patients than in non-diabetic patients with coronary artery disease. Unfortunately, too many Type 2 diabetic patients do not receive adequate treatment or do not comply with their long-term medical recommendations. PMID- 9868998 TI - Can good surrogate end-points predict the prognosis of hypertensive patients? AB - SURROGATE END-POINTS FOR PROGNOSIS OF HYPERTENSION: The identification of surrogate measures of cardiovascular risk in patients with hypertension may allow clinicians to better estimate a patient's long-term prognosis and monitor the effects of antihypertensive therapy in reducing risk and thereby reducing the cardiovascular complications of hypertension. PROGNOSTIC LIMITATIONS OF OFFICE BLOOD PRESSURE: Previous studies have shown that office blood pressure may predict the incidence of fatal and nonfatal cardiovascular complications of hypertension. However, evidence also suggests that the predictive value of office blood pressure is limited and that it does not provide accurate estimates of the changes in the cardiovascular risk profile that can occur with antihypertensive treatment. PROGNOSTIC VALUE OF 24-H AMBULATORY BLOOD PRESSURE: Cross-sectional studies have shown that 24-h average blood pressure values are more closely correlated with hypertensive target-organ damage [e.g. left ventricular hypertrophy (LVH), retinopathy, increased serum creatinine, albuminuria, and microalbuminuria] than are office blood pressure values. Although longitudinal evidence of the clinical relevance of 24-h ambulatory blood pressure monitoring is limited, preliminary data from a recently completed trial, the Study on Ambulatory Pressure and Lisinopril Evaluation (SAMPLE), have clearly shown the superiority of 24-h blood pressure monitoring over office readings in predicting the regression of LVH in hypertensive patients following treatment to reduce blood pressure. PMID- 9868999 TI - Regression of cardiac hypertrophy by 1,4 dihydropyridines in hypertensive patients. AB - BACKGROUND: Left ventricular hypertrophy (LVH) represents an important intermediate end-point for, for example, the progression to heart failure. The persistence or progression of LVH despite antihypertensive therapy probably reflects the persistence or activation of mechanisms that negatively affect the cardiovascular system and, consequently, long-term outcome. EFFECT OF MODERN ANTIHYPERTENSIVE AGENTS: Long-term treatment with rapid-onset (and usually short acting) dihydropyridine calcium antagonists is significantly less effective than angiotensin converting enzyme (ACE) inhibition in reducing left ventricular mass (LVM) in hypertensive patients. Both intermittent, and probably only partial, blood pressure control and an increase in sympathetic activity resulting from rapid decreases in blood pressure following dosing with such calcium antagonists may contribute to this relative ineffectiveness. In contrast, more recent studies have demonstrated that the longer acting dihydropyridines can reduce LVM as effectively as ACE inhibitors. DISTINCTIONS AMONG DIHYDROPYRIDINE CALCIUM ANTAGONISTS: Among the 1,4-dihydropyridines, drugs such as amlodipine and nifedipine in the gastrointestinal therapeutic system (GITS) maintain good blood pressure control over the full 24-h dosing period and do not cause dose-related increases in sympathetic activity. In contrast, extended-release felodipine has been shown to provide only intermittent blood pressure control, still leading to sympathetic activation. Notably, during short periods of noncompliance, blood pressure control is maintained with intrinsically long-acting agents such as amlodipine but not with slow-release formulations of short-acting agents such as nifedipine GITS. CONCLUSIONS: It is possible that the rate of onset and duration of action of various dihydropyridines may be pivotal factors in determining their effects on cardiovascular outcomes. Thus, the least beneficial dihydropyridines may be rapid-onset, short-acting agents, such as nifedipine capsules, and the most beneficial may be the slow-onset, long-acting agents such as amlodipine. PMID- 9869000 TI - Modern antihypertensive treatment and the progression of renal disease. AB - BACKGROUND: In animal models of hypertension, the resistance state of the preglomerular (afferent) and postglomerular (efferent) capillary arterioles may determine whether a particular form of antihypertensive therapy will spare the kidney from hemodynamic-mediated glomerular injury. In experimental models of renal disease with impaired autoregulation, control of systemic blood pressure is a prerequisite for normalizing glomerular capillary hydraulic pressure. CLINICAL STUDIES: In humans, effective blood pressure control reverses renal hemodynamic abnormalities in hypertensive patients, reduces microalbuminuria in essential hypertensive, nondiabetic, and diabetic renal diseases, and attenuates but does not prevent the progression of nondiabetic and diabetic renal disease. Although some researchers have concluded that angiotensin converting enzyme inhibitors are the renal protective drugs of choice, these pronouncements are not based on clinical trials correlating specific drug-mediated changes in albumin or protein excretion with the longitudinal assessment of glomerular filtration rate (GFR), permitting derivation of a slope-defining change in GFR, and/or the longitudinal assessment of renal structure (i.e. renal biopsy). Definitive clinical trials have not been reported. It is important to recognize that an elevated serum creatinine is a powerful predictor of mortality and that, in most patients, death is caused by a cardiovascular or cerebrovascular event, rather than by renal failure. CONCLUSION: Because morbidity and mortality of essential hypertension and nondiabetic or diabetic renal disease is related primarily to cardiovascular or cerebrovascular events, the antihypertensive 'drugs of choice' should be those that reduce these risks, prevent or regress target-organ damage, and optimize treatment of concomitant diseases. PMID- 9869001 TI - Vascular protection with newer antihypertensive agents. AB - ROLE OF CALCIUM: Alterations in the structure and function of small arteries may contribute to elevated blood pressure in patients with essential hypertension and the cardiovascular complications of this disease. Angiotensin II, endothelin 1, and other peptides, as well as catecholamines, may contribute to the altered structure and function of small arteries, in part through their effects on intracellular calcium. Intracellular calcium is implicated in signal transduction pathways that result in smooth muscle cell growth and contraction and in the release of endothelium-dependent agents involved in the regulation of vascular function. VASCULAR EFFECTS OF NEWER ANTIHYPERTENSIVE AGENTS: The newer antihypertensive agents that affect intracellular calcium pathways either directly or indirectly, such as calcium antagonists or angiotensin converting enzyme (ACE) inhibitors, may exert vascular protective effects, partly by regressing vascular remodeling and by correcting endothelial dysfunction. A double-blind trial comparing the effects of the ACE inhibitor cilazapril and the beta-blocker atenolol on media: lumen ratio of small gluteal subcutaneous arteries, which was significantly higher in hypertensive patients before treatment than in normotensive subjects, was corrected after treatment with cilazapril. In contrast, no improvement was seen in the media: lumen ratio of small arteries in the patients receiving atenolol, even after 2 years of treatment. In another study, treatment with the intrinsically long-acting calcium antagonist amlodipine resulted in a reduction in forearm minimal vascular resistance. Patients treated with amlodipine also showed normalized responses of forearm blood flow to nitric oxide synthase inhibition. Furthermore, patients treated with a slow-release formulation of nifedipine exhibited normal media: lumen ratio of small arteries and normal endothelium-dependent relaxation. CONCLUSIONS: These findings appear very promising; however, it remains to be demonstrated whether the apparently beneficial effects of treatment with long acting calcium antagonists and ACE inhibitors will lead to reductions in the cardiovascular morbidity and mortality associated with hypertension. PMID- 9869002 TI - In search of hypertension genes in Dahl salt-sensitive rats. AB - Quantitative trait loci in Dahl rats Genetic and crude physical mapping have yielded chromosome regions containing quantitative trait loci for blood pressure in Dahl salt-sensitive rats. So far, the molecular identities of these loci are largely unknown. Intriguing still is how these quantitative trait loci would interact with each other to achieve an overall blood pressure effect Alleles of some loci previously identified as blood pressure quantitative trait loci in other rat strains appear to be the same between Dahl salt-sensitive and salt resistant rats. Why do Dahl salt-resistant rats have low blood pressure whereas Dahl salt-sensitive rats develop high blood pressure? Recent findings With the use of congenic strains and 'double' congenics, these issues have begun to unravel. Certain quantitative trait loci exert major blood pressure effects (>20 mmHg) and each of them can be dissected as a monogenic trait Some appear to be located close to each other in the same chromosome region. Different quantitative trait loci interact epistatically to produce their combined blood pressure effects. 'Low' blood pressure alleles of one quantitative trait locus can compensate for the 'high' blood pressure alleles of other quantitative trait loci in the Dahl salt-resistant rat By integrating fine mapping and positional cloning strategies, blood pressure quantitative trait loci are being elucidated. Work in the rat may also facilitate genetic mapping of quantitative trait loci in humans. PMID- 9869003 TI - Ambulatory blood pressure monitoring and the taxonomy of hypertension. PMID- 9869004 TI - Blood pressure in relation to obesity, insulin and the hypothalamic-pituitary adrenal axis in Swedish men. AB - OBJECTIVE: Some studies have shown a clustering of obesity, insulin and hypertension. The present study was performed to further characterize these associations. SUBJECTS AND METHODS: In a population of 51-year-old men (n=284), measurements of systolic and diastolic blood pressure were analyzed in relation to general obesity (body mass index) and central obesity (waist: hip circumference ratio and abdominal sagittal diameter), and to the fasting insulin and insulin: glucose ratio as an approximation of insulin sensitivity. The regulation of diurnal cortisol secretion was examined in repeated salivary samples. RESULTS: Linear regression analysis showed that all three parameters of obesity were significantly and strongly related to both systolic and diastolic blood pressure, more powerfully than insulin, glucose and insulin sensitivity (insulin: glucose ratio). Stepwise multiple regression showed that only central obesity, measured as the abdominal sagittal diameter, remained significantly (P< 0.001), and independently of insulin and insulin sensitivity, associated with both systolic and diastolic blood pressure (beta=7.5 and 4.2, respectively). A diurnal cortisol curve with normal rhythm was associated with lower than average blood pressures (P< 0.001) but not with insulin levels or the heart rate. In contrast, a flattened diurnal cortisol curve, indicating perturbations in the activity of the hypothalamic-pituitary-adrenal axis, was directly related to blood pressures, heart rate and insulin (P<0.001), and has previously been found to be strongly associated with abdominal obesity. CONCLUSIONS: These findings suggest that general and central obesity is independently related to blood pressure, and that insulin may account for only part of this association. The activity of the hypothalamic-pituitary-adrenal axis is apparently important for blood pressure regulation, suggesting that mechanisms of the central nervous system have an impact. PMID- 9869005 TI - Anxiety and cardiovascular reactivity in the Tecumseh population. AB - OBJECTIVE: Increased cardiovascular reactivity has been proposed to be a critical mediator in the development of hypertension and cardiovascular disease. The personality factors associated with cardiovascular reactivity are still subject to debate. The studies reported here were undertaken to examine the relationship between trait anxiety and cardiovascular stress reactivity in a community-based sample (Tecumseh). DESIGN AND METHODS: All studies were carried out in an outpatient setting. Cardiovascular reactivity to isometric handgrip and mental arithmetic was assessed and recorded by automatic blood pressure monitoring in 832 subjects aged 19-41 years. Spielberger trait and state anxiety measures were collected immediately before the stressors were applied. RESULTS: No differences in baseline heart rate, systolic or diastolic blood pressure were observed across anxiety categories. There was a clear negative correlation between trait anxiety and cardiovascular reactivity to mental arithmetic. The pattern was less clear in response to isometric handgrip. CONCLUSIONS: These results suggest that individuals with high trait anxiety demonstrate reduced cardiovascular reactivity while those with low trait anxiety demonstrate increased reactivity, whereas the opposite might have been expected. PMID- 9869006 TI - Ambulatory blood pressure in the hypertensive population: patterns and prevalence of hypertensive subforms. AB - BACKGROUND: A number of clinically identifiable patterns of blood pressure elevation are apparent using ambulatory measurement Their prevalence and age and sex distribution have not been described. The purpose of this study was to describe the epidemiology of patterns of high blood pressure in a large population. DESIGN: Retrospective database analysis of referral hypertensive population. PATIENTS: Clinic and ambulatory blood pressure measurements were performed in 2092 patients with essential hypertension while they were not taking antihypertensive medication. METHODS: The patients were classified into six groups on the basis of their ambulatory blood pressure monitoring profiles: white coat hypertensives, borderline hypertensives, isolated systolic hypertensives, isolated diastolic hypertensives, combined systolic and diastolic hypertensives and nocturnal hypertensives. The categories were examined for age and sex differences. RESULTS: All patients were categorized into one of the six groups. The majority (56.2%) were systolodiastolic hypertensives, 12.9% were borderline and 10.8% were white-coat hypertensives. Isolated systolic hypertensives comprised 6.2% of the population, isolated diastolic hypertensives 6.9% and nocturnal hypertensives 7.1%. The isolated systolic hypertensives showed the greatest change with age, with a prevalence of < 5% in patients below 40 years of age, rising to almost 20% in the 70 plus age group. White-coat, isolated diastolic and borderline hypertensives showed the opposite change in prevalence, falling from younger to older age groups. Apart from a greater prevalence of white-coat hypertension in females, the patterns were largely similar between sexes. CONCLUSIONS: Ambulatory blood pressure monitoring allows the blood pressure pattern to be defined in hypertensive patients. The patterns identified here in a referral hypertensive cohort show changes in prevalence with age, and further study is required to determine the prognostic significance of hypertensive sub-forms. PMID- 9869007 TI - Changes in the circadian rhythm of blood pressure in primary aldosteronism in response to dietary sodium restriction and adrenalectomy. AB - OBJECTIVE: Recently, we found that sodium restriction restored the circadian rhythm of blood pressure from non-dippers to dippers in patients with a sodium sensitive type of essential hypertension. In the present study, we investigated the effects of sodium restriction on the circadian blood pressure rhythm in patients with primary aldosteronism, a typical sodium-sensitive form of secondary hypertension. DESIGN AND METHODS: We performed 24 h blood pressure monitoring in eight patients with primary aldosteronism due to unilateral adenoma (Conn's syndrome) during normal-sodium (7-12 g/day of NaCl) and low-sodium (1-3 g/day) diets, and after adrenalectomy. RESULTS: Sodium restriction lowered the 24 h mean arterial pressure from 116+/-14 to 109+/-12 mmHg (P< 0.01). During a normal sodium diet, there was no change in systolic, diastolic and mean arterial pressures during the night-time compared with the daytime. In contrast, during a low-sodium diet, all night-time pressure values were significantly lower than those in the daytime. After adrenalectomy, the night-time pressures in patients on a normal-sodium diet were lower than those of the daytime. The nocturnal mean arterial pressure fall was increased by sodium restriction and adrenalectomy. CONCLUSIONS: These results indicate that the circadian rhythm of blood pressure was disturbed in patients with primary aldosteronism who maintained a relatively high sodium intake. Both adrenalectomy and sodium restriction restored a nocturnal dip in blood pressure in primary aldosteronism. Therefore, sodium restriction affects the circadian blood pressure rhythm in sodium-sensitive types of hypertension, not only in primary hypertension, but also in secondary hypertension. PMID- 9869008 TI - Erythropoietin upregulates angiotensin receptors in cultured rat vascular smooth muscle cells. AB - OBJECTIVE: Plasma renin is not elevated in recombinant human erythropoietin (rhEPO)-induced hypertension but angiotensin converting enzyme inhibitors reduce blood pressure in both human and animal studies. Since rhEPO elevates renin and angiotensinogen messenger RNAs in angiotensin II target tissues such as the aorta, we explored the actions of rhEPO on renin-angiotensin system-related gene transcription of cultured rat vascular smooth muscle cells. DESIGN AND METHODS: To separate direct actions of rhEPO from those mediated secondarily by potential activation of the renin-angiotensin system, vascular smooth muscle cells were cultured with rhEPO and enalapril to inhibit the angiotensin converting enzyme and losartan to inhibit angiotensin II type 1 receptors. RESULTS: Vascular smooth muscle cells cultured with rhEPO (6-8 units/ml) demonstrated elevations (40-120%) in messenger RNAs of the renin-angiotensin system (renin, angiotensinogen, angiotensin receptor types 1 and 2) and increased levels of several messenger RNAs known to respond to angiotensin II (transforming growth factor-beta, insulin like growth factor-II, epidermal growth factor, c-fos and platelet-derived growth factor). In contrast, cells cultured in the presence of rhEPO and enalapril or losartan showed elevations of messenger RNA for only the two types of angiotensin II receptor. This increase was higher than that obtained when cells were cultured with rhEPO or either antagonist alone. The increase in specific binding of angiotensin II to cells cultured in the presence of rhEPO and enalapril or rhEPO and losartan paralleled the changes in receptor messenger RNA. CONCLUSIONS: rhEPO exerts its primary action on vascular smooth muscle cells via an increase in angiotensin receptor messenger RNA, resulting in a parallel increase in angiotensin II receptor expression. We suggest that increased receptor expression secondarily mediates the expression of other renin-angiotensin system messenger RNAs, which leads to angiotensin II-responsive gene transcription. The elevation in angiotensin II receptors, as observed in response to rhEPO, may provide a mechanism by which other forms of renin-dependent hypertension are initiated. PMID- 9869009 TI - Dissociation of blood pressure reduction from end-organ damage in TGR(mREN2)27 transgenic hypertensive rats. AB - OBJECTIVE: Since the biochemical disturbance underlying hypertension may be an important determinant of patient outcome, we compared the effects of early treatment with different antihypertensive drugs on end-organ damage in the TGR(mREN2)27 transgenic rat (REN-2). In these REN-2 rats, hypertension is primarily caused by increased activity of the tissue renin-angiotensin system. DESIGN AND METHODS: Seven-week-old REN-2 rats were either untreated or treated orally with an optimal daily dose of carvedilol (30 mg/kg), hydralazine (30 mg/kg), losartan (10 mg/kg) or quinapril (15 mg/kg). Nontransgenic littermates served as normotensive controls. After 11 weeks of treatment, we determined plasma norepinephrine concentrations, left ventricular atrial natriuretic factor messenger RNA and cardiac and vascular function and hypertrophy. RESULTS: Chronic treatment with carvedilol and hydralazine significantly decreased blood pressure to a similar level but failed to normalize it, whereas both losartan and quinapril completely normalized blood pressure. Despite a blood pressure reduction in all treatment groups, only losartan, quinapril and hydralazine preserved endothelial function, while carvedilol did not. Furthermore, losartan and quinapril prevented cardiac and medial hypertrophy. The expression of atrial natriuretic factor messenger RNA paralleled the hemodynamic changes. Plasma norepinephrine levels were normalized by losartan or quinapril but remained increased after carvedilol and hydralazine treatment. CONCLUSIONS: In REN-2 hypertensive rats, end-organ damage can be prevented by both inhibition of the angiotensin converting enzyme and blockade of the angiotensin II type 1 receptor, but not by merely lowering blood pressure. When blood pressure is not fully normalized, the effects on end-organs are clearly dissociated from the antihypertensive effects. PMID- 9869010 TI - Role of angiotensin II in hyperinsulinemia-induced hypertension in rats. AB - OBJECTIVE: To investigate the role of angiotensin II in the pathogenesis of hyperinsulinemia-induced hypertension in rats. MATERIALS AND METHODS: Chronic hyperinsulinemia was achieved by infusing insulin (3 mU/kg per min) subcutaneously by an osmotic minipump for 6 weeks. An angiotensin converting enzyme inhibitor (fosinopril, 10 mg/kg per day) was added in drinking water and the angiotensin II subtype 1 receptor antagonist losartan (3.5 microg/kg per min) was co-infused via the minipump. Control rats were administered the vehicle only. The rats were housed in individual metabolic cages and fed a sodium-controlled diet. Food and water intake and urine output were measured daily. Systolic blood pressure and heart rate were measured by the tail-cuff method twice a week. RESULTS: By the end of weeks 4 and 6 of the sustained insulin infusion, systolic blood pressure had increased significantly (P < 0.05), from 134+/-1 to 157+/-2 and 158+/-2 mmHg, respectively, and the heart rate had increased significantly (P< 0.05), from 380+/-9 to 423+/-7 and 426+/-6 beats/min, respectively. The plasma insulin concentration increased by 2-2.5 times but no significant changes in plasma glucose and triglyceride levels were noted. Concomitant treatment with fosinopril prevented the rises in systolic blood pressure and heart rate in the insulin-infused rats. When the insulin-induced hypertension had become established (systolic blood pressure increased from 132+/-3 to 155+/-2 mmHg 4 weeks after the infusion, P< 0.05 ), subsequent fosinopril or losartan treatment for 2 weeks reversed the elevated systolic blood pressure and heart rate to the control levels. There were no significant differences in water intake, urine flow, sodium gain and body weight gain between the control and the insulin infused rats. CONCLUSIONS: Angiotensin converting enzyme inhibition or angiotensin II type 1 receptor antagonism can prevent and reverse insulin-induced hypertension in rats, suggesting that angiotensin II itself or an angiotensin II dependent mechanism has an etiological influence in the pathogenesis of this hypertension model. PMID- 9869012 TI - Slower activation of insulin action in hypertension associated with obesity. AB - OBJECTIVE: To determine whether kinetic abnormalities in the onset of insulin action contribute to the insulin resistance in obesity-associated hypertension. DESIGN: We monitored the rate of increase in glucose infusion during 6 h of hyperinsulinemic (40 mU/m2 per min) euglycemic clamps in hypertensive and normotensive obese subjects. The two groups of hypertensive (n=9) and normotensive (n=9) subjects were matched for age (48+/-2 versus 45+/-5 years), sex (five males and four females versus four males and five females) and body mass index (42+/-3 versus 40+/-2 kg/m2). RESULTS: In all subjects, the glucose infusion rate required to maintain euglycemia increased progressively during the clamp studies to achieve maximal, steady-state values within the fifth hour. During the first 2 h of the clamp, mean glucose infusion rate, the traditional approach to assessing insulin sensitivity, was lower in the hypertensive than in the normotensive obese patients (2.04+/-0.13 versus 3.29+/-0.41 mg/kg per min, respectively; P < 0.05). In contrast, the maximal steady-state glucose infusion rate, calculated as the mean value during the sixth hour of clamping, was similar in the hypertensive and in the normotensive obese patients (4.48+/-0.43 versus 4.81+/-0.45 mg/kg per min, respectively; NS). The time required to reach the half maximal glucose infusion rate was greater in the hypertensive than normotensive obese patients (91+/-12 versus 38+/-5 min, respectively; P< 0.05). CONCLUSION: In obesity, hypertension was associated with a slower rate of activation of the insulin effect on glucose metabolism, whereas the maximal steady-state insulin effects were not altered by elevated blood pressure. Thus, the link between obesity and hypertension may be associated with the kinetics of onset of insulin action. PMID- 9869011 TI - Overexpression of vascular endothelin-1 and endothelin-A receptors in a fructose induced hypertensive rat model. AB - OBJECTIVE: To examine the temporal relationship between hyperinsulinemia and hypertension in the fructose-hypertensive rat model and to study the function of endothelin-1 (ET-1) in fructose-induced hypertension. DESIGN: Since ET-1 induces insulin resistance in conscious rats, we tested the hypothesis that both hyperinsulinemia and hypertension developed in the fructose-hypertensive rat model might be the sequelae of an elevated tissue content of ET-1 and ET(A) receptors. MATERIALS AND METHODS: Systolic hypertension was induced within 3 weeks in male Sprague-Dawley rats fed on a fructose-rich diet. After continual monitoring of blood pressure and plasma insulin concentrations, the animals were killed at the end of experiment to determine plasma levels of ET-1, the contractile response of aortic rings to ET-1, and ET-1 and ET(A) receptor gene expressions. In a separate experiment, BQ-610 was administered to lower the effect of ET-1 in rats with fructose-induced hypertension. RESULTS: Compared with control rats given normal chow, the fructose-fed rats developed systolic hypertension after 3 weeks of the diet (127+/-3.7 versus 110+/-5.5 mmHg, P < 0.01) and hyperinsulinemia both before (1 07.1+/-32.5 versus 48.5+/-14.3 pmol/l, P < 0.005) and after (96.6+/-63.7 versus 50.4+/-5.6 pmol/l, P< 0.05) they became hypertensive. Although plasma ET-1 levels did not differ between the rat groups, aortic ring contraction-concentration curves, indicating vessel contractility in response to ET-1, were significantly greater in these rats than in controls (F1,72 = 12.34, P< 0.00077). Messenger RNA extracted from the tail arteries and blotted with both ET-1 and ET(A) probes showed that fructose-fed rats had greater ET-1 and ET(A)-receptor gene expression than control rats. Concomitant administration of BQ-610 to rats fed on a fructose diet significantly reduced the hypertension. Conclusions These findings suggest that elevated vascular expression of ET-1 and ET(A) receptor genes may mediate the development of hypertension and hyperinsulinemia in rats fed a fructose-rich diet PMID- 9869013 TI - Effect of chronic angiotensin converting enzyme inhibition on sympathetic nerve traffic and baroreflex control of the circulation in essential hypertension. AB - BACKGROUND: Human studies have shown that the blood pressure lowering effects of angiotensin converting enzyme inhibitors are accompanied by a reduction in plasma norepinephrine levels. Whether this is due to central or peripheral mechanisms is unknown, however. OBJECTIVE: To evaluate the effects of chronic interference with the renin-angiotensin system on sympathetic nerve traffic and baroreflex control of vagal and adrenergic cardiovascular drive. PATIENTS AND METHODS: In 18 untreated mild to moderate essential hypertensive patients aged 48.5+/-1.9 years (mean+/-SEM), we measured mean arterial pressure (Finapres), heart rate (electrocardiogram), plasma renin activity (radioimmunoassay), plasma norepinephrine (high-performance liquid chromatography) and postganglionic muscle sympathetic nerve activity (microneurography at a peroneal nerve). In nine patients, measurements were performed before and after 2 months of oral administration of lisinopril (10 mg/day), while in the remaining nine patients they were performed before and after a 2 month observation period, without the drug administration. Measurements were performed at rest and during baroreflex stimulation and deactivation elicited by stepwise intravenous infusions of phenylephrine and nitroprusside, respectively. RESULTS: Lisinopril induced a marked increase in plasma renin activity (from 1.1+/-0.2 to 6.4+/-1.3 ng/ml per h, P< 0.01) and a reduction in mean arterial pressure (from 109.6+/-3.1 to 98.7+/ 2.9 mmHg, P < 0.01) without affecting the heart rate. Plasma norepinephrine and muscle sympathetic nerve activity values were not significantly different before and after lisinopril treatment (plasma norepinephrine values changed from 290.4+/ 39.2 to 308.1+/-67.1 pg/ml; muscle sympathetic nerve activity changed from 56.4+/ 5.3 to 50.6+/-6.6 bursts/100 heart beats). Neither the sympathoinhibitory nor the sympathoexcitatory responses to phenylephrine and nitroprusside were affected by lisinopril, nor the concomitant bradycardia and tachycardia. The curves relating mean arterial pressure to heart rate and muscle sympathetic nerve activity values during baroreceptor manipulation were shifted to the left, indicating a resetting of the baroreflex to the lower blood pressure values achieved during treatment. CONCLUSIONS In essential hypertension, sympathetic nerve traffic is not affected by chronic angiotensin converting enzyme inhibitor treatment that effectively interferes with the renin-angiotensin system and lowers the elevated blood pressure. The baroreflex ability to modulate heart rate and central sympathetic outflow is also unaffected. These data argue against the existence of a central sympathoexcitatory effect of angiotensin II in this condition. They also indicate that antihypertensive treatment with an angiotensin converting enzyme inhibitor preserves autonomic reflex control, with favorable consequences for cardiovascular homeostasis. PMID- 9869014 TI - Interaction of bradykinin and angiotensin-(1-7) in the central modulation of the baroreflex control of the heart rate. AB - OBJECTIVE: Previous studies have shown that angiotensin-(1-7) potentiates the vascular actions of bradykinin. In the present study, we evaluated the interaction of bradykinin and angiotensin-(1-7) in the central modulation of baroreflex control of the heart rate. MATERIALS AND METHODS: Blood pressure and reflex bradycardia, elicited by intravenous injection of phenylephrine, were evaluated in conscious male Wistar rats before and at the end of 1 h of an intracerebroventricular infusion of angiotensin-(1-7) at 0.5 or 1.0 microg/h combined with bradykinin at 2.5 microg/h; or angiotensin-(1-7) at 2.0 microg/h combined with bradykinin at 4.0 microg/h; or angiotensin-(1-7) alone at 2.0 or 4.0 microg/h; or bradykinin alone at 4.0 or 8.0 microg/h; or saline at 8 microl/h. In addition, baroreflex bradycardia was evaluated before and at the end of 1 and 2 h of intracerebroventricular infusion of angiotensin-(1-7) at 4 microg/h for 2 h; or saline at 8 microl/h in the first hour followed by HOE 140 at 90 ng/h in the second hour; or angiotensin-(1-7) at 4 microg/h in the first hour followed by angiotensin-(1-7) at 4 microg combined with HOE 140 at 90 ng/h in the second hour; or HOE 140 at 90 ng/h in the first hour followed by HOE 140 at 90th ng/h combined with angiotensin-(1-7) at 4 microg/h in the second hour; or saline at 8 microl/h for 2 h. RESULTS: The intracerebroventricular infusion of angiotensin-(1-7) or bradykinin alone required a dose of 4.0 and 8.0 microg/h, respectively, to facilitate baroreflex control of the heart. However, a simultaneous infusion of these peptides at subeffective rates was able to produce a significant increase in baroreflex sensitivity. In addition, the facilitation of the baroreflex control of the heart rate induced by angiotensin-(1-7) at 4.0 microg/h was inhibited by HOE 140. CONCLUSIONS: These results suggest that centrally, bradykinin and angiotensin-(1-7) can interact in order to modulate baroreflex control of the heart rate. In addition, our data indicate that the central modulatory effect of angiotensin-(1-7) on the baroreflex is mediated, at least in part, by the release of kinins. PMID- 9869015 TI - Structural association between the carotid artery and the left ventricle in a general population in Northern Italy: the Vobarno study. AB - OBJECTIVE: To evaluate the interrelationships between structural alterations in the carotid arteries and left ventricular geometric patterns in a middle-aged general population. DESIGN AND METHODS: We studied 223 untreated subjects (106 females, 117 males; aged 48-64 years) living in a small town in Northern Italy (Vobarno, Brescia), using a cross-sectional design. The left ventricular mass index was assessed by echocardiography, while the intima-media thickness and the occurrence of plaque were evaluated by ultrasound. Blood pressure was determined by clinic measurements and by 24 h noninvasive ambulatory blood pressure monitoring. RESULTS: According to the presence of left ventricular hypertrophy (left ventricular mass index > 50 g/m2.7 in males and > 47 g/m2.7 in females) or concentric left ventricular remodelling (normal left ventricular mass index and relative wall thickness > 0.42), the subjects were divided into four groups: 124 subjects with normal left ventricular geometry, 73 subjects with left ventricular hypertrophy (55 with eccentric and 18 with concentric hypertrophy) and 26 subjects with concentric remodelling. The common carotid intima-media thickness and cross-sectional area were significantly greater in the subjects with concentric left ventricular hypertrophy (analysis of variance, P< 0.05) than in those with normal left ventricular geometry. A significantly higher number of plaques was observed in subjects with concentric and eccentric left ventricular hypertrophy. CONCLUSIONS: In a general population of unselected middle-aged subjects, the presence of concentric left ventricular hypertrophy was associated with an increase in intima-media thickness and with the presence of plaque in the carotid arteries, possibly contributing to the worse prognosis observed in this group of patients. PMID- 9869016 TI - Effects of chronic adenosine uptake blockade on adrenergic responsiveness and left ventricular chamber function in pressure overload hypertrophy in the rat. AB - BACKGROUND: Increased sympathetic activity contributes to the progression of heart failure. Adenosine counteracts sympathetic activity by inhibition of presynaptic norepinephrine release and attenuation of the metabolic and contractile responses to beta-adrenergic stimulation. In this study, we tested the hypothesis that the adenosinergic effects (uptake blockade) of dipyridamole may retard the progression of pressure overload hypertrophy in the rat. METHODS AND RESULTS: To verify that the administration of dipyridamole increases myocardial adenosine levels in the rat, epicardial adenosine concentrations were measured from 12 isolated, perfused rat hearts exposed to 10(-7) and 10(-6) mol/l dipyridamole. Adenosine concentrations were increased with both doses of dipyridamole. Also, 9 weeks of dipyridamole treatment resulted in decreased sensitivity to the adenosine A1-receptor agonist, 2-chloro-N6-cyclopentyl adenosine, suggesting that dipyridamole increases adenosine levels in the intact rat. In the second part of the study, rats were divided into either abdominal aortic-banded or sham-operated groups and were treated with either dipyridamole or saline. After 9 weeks of treatment, two-dimensional Doppler echocardiographic studies were performed and the adrenergic responsiveness to 10(-8) mol/l isoproterenol was assessed in vitro. The saline-treated banded group demonstrated concentric left ventricular hypertrophy, abnormal diastolic filling, increased wet lung weights and attenuation of adrenergic responsiveness. In contrast, the dipyridamole-treated banded rats exhibited more concentric geometry (higher relative wall thickness with similar left ventricular mass), normal left ventricular filling characteristics and preserved adrenergic responsiveness. Systolic left ventricular chamber and myocardial function, as assessed by stress endocardial and midwall shortening relationships, were not significantly altered by banding or dipyridamole treatment. CONCLUSIONS: Dipyridamole treatment prevented the development of abnormal left ventricular chamber filling, preserved adrenergic responsiveness and appeared to attenuate detrimental chamber remodeling in rats with pressure overload hypertrophy. PMID- 9869017 TI - Comparison of active treatment and placebo in older Chinese patients with isolated systolic hypertension. Systolic Hypertension in China (Syst-China) Collaborative Group. AB - BACKGROUND: Isolated systolic hypertension occurs in around 8% of Chinese people aged 60 years or older. In 1988, the Systolic Hypertension in China (Syst-China) Collaborative Group started to investigate whether active treatment could reduce the incidence of stroke and other cardiovascular complications in older patients with isolated systolic hypertension. METHODS: All patients were initially started on masked placebo. After stratification for centre, sex and previous cardiovascular complications, alternate patients (n=1253) were assigned nitrendipine at 10-40 mg daily, with the addition of captopril at 12.5-50.0 mg daily or hydrochlorothiazide at 12.5-50.0 mg daily, or both, if a sufficient blood pressure fall was not obtained. In the remaining 1141 control patients, matching placebos were administered similarly. RESULTS: At entry, sitting blood pressure averaged 170.5 mmHg systolic and 86.0 mmHg diastolic, age averaged 66.5 years and total serum cholesterol was 5.1 mmol/l. After 2 years of follow-up, sitting systolic and diastolic blood pressures had fallen by 10.9 mmHg and 1.9 mmHg in the placebo group and by 20.0 mmHg and 5.0 mmHg in the active treatment group. The intergroup differences were 9.1 mmHg systolic (95% confidence interval 7.6-10.7 mmHg ) and 3.2 mmHg diastolic (95% confidence interval 2.4-4.0). Active treatment reduced total strokes by 38% (from 20.8 to 13.0 endpoints per 1000 patient-years, P=0.01), all-cause mortality by 39% (from 28.4 to 17.4 endpoints per 1000 patient-years, P=0.003), cardiovascular mortality by 39% (from 15.2 to 9.4 endpoints per 1000 patient-years, P=0.03), stroke mortality by 58% (from 6.9 to 2.9 endpoints per 1000 patient-years, P=0.02), and ail fatal and nonfatal cardiovascular endpoints by 37% (from 33.3 to 21.4 endpoints per 1000 patient years, P=0.004). CONCLUSIONS: Antihypertensive treatment prevents stroke and other cardiovascular complications in older Chinese patients with isolated systolic hypertension. Treatment of 1000 Chinese patients for 5 years could prevent 55 deaths, 39 strokes or 59 major cardiovascular endpoints. PMID- 9869018 TI - Antihypertensive efficacy of lercanidipine at 2.5, 5 and 10 mg in mild to moderate essential hypertensives assessed by clinic and ambulatory blood pressure measurements. Multicenter Study Investigators. AB - OBJECTIVE: To evaluate the antihypertensive effect of lercanidipine once a day at three different doses (2.5, 5 and 10 mg) by clinic and ambulatory blood pressure. METHODS: After 3 weeks of a placebo run-in, 243 mild to moderate essential hypertensives (mean+/-SD age 51+/-8 years) were randomly allocated to lercanidipine at 2.5, 5 or 10 mg or a placebo for 4 weeks, in a double-blind parallel-group design. At the end of each period, supine clinic blood pressure (standard sphygmomanometry) and 24 h ambulatory blood pressure (Spacelabs 90207) were measured. The duration and homogeneity of the antihypertensive effect of the active drug compared with placebo over 24 h was evaluated by calculating the smoothness index, the ratio of the mean of the 24 hourly blood pressure changes to the corresponding SD. The higher the smoothness index, the greater and the smoother is the antihypertensive effect of a drug over the 24 h. RESULTS: In 211 patients with valid clinic blood pressure data at the end of treatment, larger systolic/diastolic blood pressure reductions were found in the 5 mg (10+/-12/8+/ 6 mmHg; P< 0.05 versus placebo, diastolic blood pressure only) and the 10 mg (12+/-11/9+/-7 mmHg; P < 0.05 versus placebo, both pressures) lercanidipine groups than in the placebo (5+/-11/4+/-8 mmHg) and 2.5 mg lercanidipine (7+/-12/6 +/-7 mmHg) groups. In 105 patients with complete 24 h ambulatory blood pressure recordings, there were significantly (P< 0.05 versus placebo) larger reductions in the 10 mg (9+/-7/7+/-5 mmHg) than the 2.5 mg (1+/-10/1+/-6 mmHg) and placebo (2+/-6/1+/-4 mmHg) groups. The reduction in 24 h blood pressure with 5 mg lercanidipine (6+/-7/4+/-5 mmHg) was significantly greater compared with placebo for diastolic pressure only, and when hourly average blood pressure changes were considered, this reduction did not extend to the final hours of the dosing interval. No significant changes in the clinic or 24 h heart rate were induced by placebo or lercanidipine. The smoothness index was significantly (P< 0.05) lower for 2.5 mg lercanidipine and placebo (0.2+/-0.5 and 0.3+/-0.7 for systolic and 0.1+/-0.4 and 0.2+/-0.7 for diastolic blood pressure) than for the 5 and 10 mg doses (0.7+/-1 and 1+/-0.7 for systolic and 0.7+/-1 and 1+/-0.9 for diastolic blood pressure). CONCLUSIONS: At a dose of 10 mg, lercanidipine had a significant and durable antihypertensive effect over 24 h, but at 5 mg, the effect was less consistent and did not last 24 h. There was no clinically relevant reduction in clinic or ambulatory blood pressure with 2.5 mg lercanidipine, and the effect was superimposable on that of placebo. PMID- 9869019 TI - Quality of life and calcium channel blockade with nifedipine GITS versus amlodipine in hypertensive patients in Spain. Gastrointestinal Therapeutic System. AB - OBJECTIVE: Compliance with hypertension treatment is affected by treatment related factors (complexity, side effects), efficacy and compound-specific effects that impact on quality of life. This study examined the differences in quality of life produced by two once-daily calcium channel blockers using different delivery systems: nifedipine gastrointestinal therapeutic system (GITS) and amlodipine. DESIGN: This was a double-blind, double-dummy, randomized clinical trial comparing nifedipine GITS (30 mg) and amlodipine (5 mg) for 24 weeks following a placebo run-in. Clinical, laboratory evaluations and quality-of life data were assessed at screening, baseline randomization and three times during active therapy. SETTING: The study was conducted in 13 medical clinics in Spain. PATIENTS: The sample comprised 430 screened and 356 randomized patients with mild to moderate hypertension (diastolic blood pressure 95-114 mmHg). MAIN OUTCOME MEASURES: Change in systolic and diastolic blood pressure and in health related quality of life were the main outcome measures. RESULTS: There were no significant differences between active treatment groups in the blood pressure changes (systolic blood pressure: nifedipine GITS -15.5 mmHg; amlodipine -15.7 mmHg). Spontaneous adverse events consistent with calcium channel blockage were not different. The nifedipine GITS group improved in all quality-of-life measures except Sexual Symptom Distress and showed a significantly greater improvement than amlodipine in overall Quality of Life (P< 0.05), General Perceived Health (P < 0.026) and its subscale Vitality (P < 0.019). The amlodipine group declined in overall Quality of Life, General Perceived Health, Vitality and Sleep Disturbance, and significantly in Sexual Symptom Distress (P < 0.045). However, this group improved in self-reported Cognitive Functioning (P=0.036), Mental Acuity (P < 0.005) and Detachment/disorientation (P=0.01). CONCLUSIONS: These results suggest compound-specific effects on quality of life that may be due to differences in the delivery system. Nifedipine GITS is short-acting (2 h half life) and is delivered continuously over a 24 h period, while amlodipine has a half-life of 40 h, which may produce more sustained low-level effects. While a more beneficial profile was observed for nifedipine, amlodipine demonstrated potential positive effects on cognitive functioning. PMID- 9869020 TI - Evaluation and management of the "new" lymphoma entities: mantle cell lymphoma, lymphoma of mucosa-associated lymphoid tissue, anaplastic large-cell lymphoma, and primary mediastinal B-cell lymphoma. AB - Non-Hodgkin's lymphomas (NHL) represent a major health problem worldwide, and incidence has been on the rise continuously for the last few decades. It is estimated that approximately 55,000 new cases of NHL will be diagnosed in the United States in 1998 and that slightly fewer than 25,000 patients will die of treatment failure or recurrent disease. The rising incidence of NHL is related not only to the acquired immunodeficiency syndrome epidemic but to also a steady increase in the number of cases diagnosed in older patients without immunosuppression. The new pathologic classification of NHL (revised European American lymphoma classification, REAL) developed by the International Lymphoma Study Group (ILSG) is already resulting in more accurate disease-specific epidemiologic and clinical investigations. These studies have brought a new awareness of the existence and the relative prevalence of discrete NHL subtypes that appear to predominate among patients in different populations according to age, sex, geographic distribution, and predisposing conditions. This developing database has also the potential to result in the discovery of specific environmental causes, predisposing genetic factors, and therapeutic approaches. Some of the entities defined in the REAL classification, such as follicular lymphomas, diffuse B large-cell lymphomas, and T-cell lymphoblastic lymphomas, were already well described in the older classification systems (Kiel and Working Formulation). Others, such as mantle cell lymphoma, (MCL) anaplastic large-cell lymphoma (ALCL), lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma), and primary mediastinal B-cell lymphoma (PMBCL) are relatively new members of the family, and accurate data on their clinicopathologic features and natural histories have only recently begun to emerge. This review presents in detail the most recent data on the clinical presentation of, diagnostic evaluation of, and treatment options for the most common of the new NHL entities: MCL, MALT lymphoma, CD30+ (Ki-1+) ALCL, and PMBCL. These four entities combined represent approximately 20% of all cases of NHL and exemplify well the broad clinicopathologic spectrum of NHL and the diagnostic and therapeutic challenges facing those who care for patients affected by these conditions. PMID- 9869021 TI - Commentary: exciting new developments in fast neutron cross sections and dosimetry. PMID- 9869022 TI - Experimental double-differential cross sections and derived kerma factors for oxygen at incident neutron energies from reaction thresholds to 65 MeV. AB - The double-differential cross sections (energy spectra) for the (n, px), (n, dx), (n, tx) and (n, rx) reactions on oxygen have been measured for nine incident neutron energies in the range 25 to 65 MeV at lab angles between 20 degrees and 160 degrees in steps of 10 degrees. From these measurements, the energy differential cross sections have been determined and consequently the partial and total kerma factors. Based on the obtained experimental partial kerma factors in the incident neutron energy range 25-65 MeV, a procedure is proposed for the extrapolation of these values to the reaction threshold energy of each measured reaction channel. Results of the experimental double-differential, energy differential and total cross sections are presented. The deduced partial and total kerma factors of the present work are compared with results of previous measurements and theoretical predictions. PMID- 9869023 TI - Direct determination of kerma for a d(48.5) + Be therapy beam. AB - An experimental determination of the neutron kerma ratio between muscle tissue and A-150 plastic was performed at the newly commissioned d(48.5)+ Be therapy facility in Detroit. Low-pressure proportional counters with separate walls made from A-150 plastic, graphite, zirconium oxide and zirconium served to measure ionization yield spectra. The absorbed dose in the wall of each counter was determined and rendered the A-150 and carbon kerma directly, whilst that for oxygen was deduced from differences between the matched metal oxide and metal pair. This enabled the evaluation of an effective kerma ratio as a function of radiation field size and hydrogenous filtration. Although filtration was observed to harden the beam, the application of a single kerma ratio for the various irradiation conditions investigated was found to be appropriate. A neutron kerma ratio of 0.90+/-0.03 was assessed for the Detroit facility, which is lower at the 1sigma level than the 0.95 currently recommended in the dosimetry protocol for high-energy neutron beams. PMID- 9869024 TI - Carbon beam dosimetry intercomparison at HIMAC. AB - To verify international uniformity in carbon beam dosimetry, an intercomparison programme was carried out at the heavy ion medical accelerator (HIMAC). Dose measurements with ionization chambers were performed for both unmodulated and 6 cm modulated 290 MeV/nucleon carbon beams. Although two different dosimetry procedures were employed, the evaluated values of absorbed dose were in good agreement. This comparison established a common framework for ionization chamber dosimetry between two different carbon beam therapy facilities. PMID- 9869025 TI - A simple method for matching electron beams in radiotherapy. AB - Many solutions have been proposed to solve the problem of 'hot' and 'cold' spots in the junction between abutting electron fields. Although some of these methods have proved satisfactory, the designs of the modifications to the applicators are generally applicator dependent and involve measuring data for individual fields. An idea which was originally proposed as a solution to a different problem is resurrected here because it happens to solve the beam-matching problem and because it is very simple to apply. The idea is to cover the end of the electron applicator completely with a slab of plastic, called a 'spoiler'. Formulae are proposed and tested for estimating the penumbra broadening and the extent of over or underdosing at field junctions. The technique is applicator independent, and no extra beam-data measurements need be made for computer treatment planning of matching fields. The price paid for such simplicity is that consideration needs to be given to leading-off the other borders of the treatment fields because of the increased penumbra widths. PMID- 9869026 TI - Monte Carlo calculations of electron beam output factors for a medical linear accelerator. AB - The purpose of this study was to investigate the application of the Monte Carlo technique to the calculation and analysis of output factors for electron beams used in radiotherapy. The code EGS4/BEAM was used to obtain phase-space files for 6, 12 and 20 MeV clinical electron beams from a scattering-foil linac (Varian Clinac 2100C) for a clinically representative range of applicator and square or rectangular insert combinations. The source-to-surface distance used was 100 cm. The field sizes ranged from 1 x 1 cm2 to 20 x 20 cm2. These phase-space files were analysed to study the intrinsic beam characteristics and used as source input for relative dose and output factor computations in homogeneous water phantoms using the code EGS4/DOSXYZ. The calculated relative central-axis depth dose and transverse dose profiles at various depths of clinical interest agreed with the corresponding measured dose profiles to within 2% of the maximum dose. Calculated output factors for the fields studied agreed with measured output factors to about 2%. This demonstrated that for the Varian Clinac 2100C linear accelerator, electron beam dose calculations in homogeneous water phantoms can be performed accurately at the 2% level using Monte Carlo simulations. PMID- 9869027 TI - Combined experimental and Monte Carlo verification of 137Cs brachytherapy plans for vaginal applicators. AB - Dose rates in a phantom around a shielded and an unshielded vaginal applicator containing Selectron low-dose-rate 137Cs sources were determined by experiment and Monte Carlo simulation. Measurements were performed with thermoluminescent dosimeters in a white polystyrene phantom using an experimental protocol geared for precision. Calculations for the same set-up were done using a version of the EGS4 Monte Carlo code system modified for brachytherapy applications into which a new combinatorial geometry package developed by Bielajew was recently incorporated. Measured dose rates agree with Monte Carlo estimates to within 5% (1 SD) for the unshielded applicator, while highlighting some experimental uncertainties for the shielded applicator. Monte Carlo calculations were also done to determine a value for the effective transmission of the shield required for clinical treatment planning, and to estimate the dose rate in water at points in axial and sagittal planes transecting the shielded applicator. Comparison with dose rates generated by the planning system indicates that agreement is better than 5% (1 SD) at most positions. The precision thermoluminescent dosimetry protocol and modified Monte Carlo code are effective complementary tools for brachytherapy applicator dosimetry. PMID- 9869028 TI - Dose perturbation caused by high-density inhomogeneities in small beams in stereotactic radiosurgery. AB - The influence of high-density tissue heterogeneities in small-diameter beams used in stereotactic radiosurgery has been investigated. Dose perturbation immediately behind aluminium sheets, used to simulate a high-density tissue inhomogeneity such as bone, was studied in a solid water phantom. Dose reduction factors (DRFs), which are the ratios of the dose in the presence of the inhomogeneity to dose in a uniform density solid water phantom, were measured with a diamond detector for three thicknesses of aluminium. DRFs exhibit dependence on both the inhomogeneity thickness and the beam diameter. The DRF decreases with inhomogeneity thickness. The DRF initially decreases with increase in the beam diameter from 12.5 to 25 mm. For fields greater than 25 mm, the DRFs are nearly constant. The commonly used algorithms such as the TAR ratio method underestimate the magnitude of the measured effect. A good agreement between these measurements and Monte Carlo calculations is obtained. The influence of the high-density inhomogeneity on the tissue maximum ratio (TMR) was also measured with the inhomogeneity at a fixed depth dmax from the entrance surface. The TMR is reduced for all detector-inhomogeneity distances investigated. The dose build-up phenomenon observed in the presence of low-density air inhomogeneity is absent in the presence of a high-density inhomogeneity. The beam width (defined by 50% dose points) immediately beyond the inhomogeneity is unaffected by the high-density inhomogeneity. However, the 90%-10% and 80%-20% dose penumbra widths and the dose outside the beam edge (beyond the 50% dose point) are reduced. This reduction in dose outside the beam edge is caused by the reduced range of the secondary radiation (photons and electrons) in the high-density medium. PMID- 9869029 TI - A model for the time-dependent thermal distribution within an iceball surrounding a cryoprobe. AB - The optimal cooling parameters to maximize cell necrosis in different types of tissue have yet to be determined. However, a critical isotherm is commonly adopted by cryosurgeons as a boundary of lethality for tissue. Locating this isotherm within an iceball is problematic due to the limitations of MRI, ultrasound and CT imaging modalities. This paper describes a time-dependent two dimensional axisymmetric model of iceball formation about a single cryoprobe and extensively compares it with experimental data. Thermal histories for several points around a CRYOprobe are predicted to high accuracy (5 degrees C maximum discrepancy). A realistic three-dimensional probe geometry is specified and cryoprobe temperature may be arbitrarily set as a function of time in the model. Three-dimensional temperature distributions within the iceball, predicted by the model at different times, are presented. Isotherm locations, as calculated with the infinite cylinder approximation, are compared with those of the model in the most appropriate region of the iceball. Infinite cylinder approximations are shown to be inaccurate when applied to this commercial probe. Adopting the infinite cylinder approximation to locate the critical isotherm is shown to lead the user to an overestimate of the volume of target tissue enclosed by this isotherm which may lead to incomplete tumour ablation. PMID- 9869030 TI - A theoretical comparison of energy sources--microwave, ultrasound and laser--for interstitial thermal therapy. AB - A number of heating sources are available for minimally invasive thermal therapy of tumours. The purpose of this work was to compare, theoretically, the heating characteristics of interstitial microwave, laser and ultrasound sources in three tissue sites: breast, brain and liver. Using a numerical method, the heating patterns, temperature profiles and expected volumes of thermal damage were calculated during standard treatment times with the condition that tissue temperatures were not permitted to rise above 100 degrees C (to ensure tissue vaporization did not occur). Ideal spherical and cylindrical applicators (200 microm and 800 microm radii respectively) were modelled for each energy source to demonstrate the relative importance of geometry and energy attenuation in determining heating and thermal damage profiles. The theoretical model included the effects of the collapse of perfusion due to heating. Heating patterns were less dependent on the energy source when small spherical applicators were modelled than for larger cylindrical applicators due to the very rapid geometrical decrease in energy with distance for the spherical applicators. For larger cylindrical applicators, the energy source was of greater importance. In this case, the energy source with the lowest attenuation coefficient was predicted to produce the largest volume of thermally coagulated tissue, in each tissue site. PMID- 9869031 TI - Initial recombination in a parallel-plate ionization chamber exposed to heavy ions. AB - For exact determination of absorbed dose in heavy-ion irradiation fields which are used in radiation therapy and biological experiments, ionization chambers have been characterized with defined heavy-ion beams and correction factors. The LET (linear energy transfer) dependence of columnar recombination in a parallel plate ionization chamber has been examined. Using 135 MeV/u carbon and neon beams, the ion collection efficiency was measured for several gases (air, carbon dioxide, argon and tissue-equivalent gas). 95 MeV/u argon beams and 90 MeV/u iron beams were also used for measurements of columnar recombination in air. As expected by Jaffe theory, the inverse of the ratio of the ionization charge to the saturated ionization charge had a linear relationship with the inverse of the electric field strength in the region below 0.002 V(-1) cm. The gradient of the line increases as the LET of the heavy ions increases. A strong LET dependence of the gradient was observed in air and carbon dioxide. The LET dependence was not observed in tissue-equivalent gas, nitrogen or argon. The exact depth-dose distribution of the heavy-ion beam was obtained by this correction of the initial recombination effect for the collected ionization charge. The columnar recombination in air was analysed using Jaffe theory; the obtained parameter b (a track radius) should be in the range between 0.001 cm and 0.005 cm, whereas the value obtained by Jaffe is 0.00179 cm. The value of the parameter b should increase as the LET of the heavy-ion beam increases in order to reproduce the experimental values of the initial recombination. PMID- 9869032 TI - Radiance modelling using the P3 approximation. AB - Light dosimetry is an essential component of effective photodynamic therapy (PDT) of tumours. Present PDT light dosimetry techniques rely on fluence-based models and measurements. However, in a previous paper by Barajas et al, radiance-based light dosimetry was explored as an alternative approach. Although successful in demonstrating the use of Monte Carlo (MC) simulations of radiance in tissue optical characterization, the MC proved time consuming and impractical for clinical applications. It was proposed that an analytical solution to the transport equation for radiance would be desirable as this would facilitate and increase the speed of tissue characterization. It has been found that the P3 approximation is one such potential solution. Radiance and fluence expressions based on the P3 approximation were used to optically characterize an Intralipid based tissue phantom of varying concentration of scatterer (Intralipid) and absorber (methylene blue) using a plane wave illuminated, semi-infinite medium geometry. The results obtained compare favourably with the Grosjean approximation of fluence (a modified diffusion theory) using the same optical parameters (mu(a), mu(s), g). The results illustrate that radiance-based light dosimetry is a viable alternative approach to tissue characterization and dosimetry. It is potentially useful for clinical applications because of the limited number of invasive measurements needed and the speed at which the tissue can be characterized. PMID- 9869033 TI - Applicability of the Kubelka-Munk theory for the evaluation of reflectance spectra demonstrated for haemoglobin-free perfused heart tissue. AB - Reflectance spectrometry is a useful tool for studying in vivo kinetic changes in the oxygen saturation of haemoglobin and myoglobin as well as the redox state of cytochromes. A method is given which allows the quantification of tissue reflectance spectra using multicomponent analysis. This method utilizes the Kubelka-Munk theory for modelling the measured tissue spectra. To test this approach, reflectance spectra of a haemoglobin-free perfused guinea pig heart were measured by a fast scanning spectrophotometer (100 spectra/s, spectral resolution 1.0 nm) and evaluated using the component absorbance spectra measured separately. A relative mean spectral residual error of 0.15% was achieved by least-squares fitting. Using statistical error propagation, oxygenation of myoglobin is obtained within a relative precision of 1%, and the redox state of cytochromes aa3 and c are determined simultaneously within a margin of 3%; the results for the redox-state of cytochrome b, however, are less precise. Special component error functions are presented to provide a reliability measure for the concentration prediction using this multicomponent assay. The consistency of the theory and the component absorptivity data is tested by regressing the actual concentrations obtained for each of the redox pair components during the various states of tissue oxygenation. A method is described for the recognition and reduction of systematic errors. PMID- 9869034 TI - Use of a priori information in estimating tissue resistivities--a simulation study. AB - Accurate estimation of tissue resistivities in vivo is needed to construct reliable human body volume conductor models in solving forward and inverse bioelectric field problems. The necessary data for the estimation can be obtained by using the four-electrode impedance measurement technique, usually employed in electrical impedance tomography. In this study, a priori geometrical information with statistical properties of regional resistivities and linearization error as well as instrumentation noise has been incorporated into a new resistivity estimation algorithm which is called a statistically constrained minimum mean squares error estimator (MiMSEE) to improve estimation accuracy. MiMSEE intakes geometrical information from the image which is obtained by using a high resolution imaging modality. This study is an extension of earlier work by Eyuboglu et al and obtains simulated measurements from two numerical models containing five and six regions on a background region. Also, estimations are repeated by using up to eight multiple current electrode pairs, in order to observe the effect of estimation performance while increasing the number of measurements up to 96. The results are compared with a conventional least squares error estimator (LSEE) which is used in one-pass algorithms. It is shown that the MiMSEE estimation error is up to 27 times smaller than the LSEE error which is realized for a small, high-contrast region, for example the aorta. In estimating the regional resistivities, the MiMSEE algorithm requires 25.8 (for the five region resistivity distribution) and 22.2 (for the six-region resistivity distribution) times more computational time than the LSEE. This gap between the computational times of the two algorithms decreases as the number of regions increases. PMID- 9869035 TI - Tomotherapeutic portal imaging for radiation treatment verification. AB - In their tomotherapy concept Mackie and co-workers proposed not only a new technique for IMRT but also an appropriate and satisfactory method of treatment verification. This method allows both monitoring of the portal dose distribution and imaging of the patient anatomy during treatment by means of online CT. This would enable the detection of inaccuracies in dose delivery and patient set-up errors. In this paper results are presented showing that a single electronic portal imaging device (EPID) could deliver all data necessary to establish such a complete verification system for tomotherapy and even other IMRT techniques. Consequently it has to be shown that it is able to record both the low-intensity photon fluences encountered in tomographic imaging and the intense photon transmission of each treatment field. The detector under investigation is a video based EPID, the BIS 710 (manufactured by Wellhofer Dosimetrie, Schwarzenbruck, Germany). To examine the suitability of the BIS for CT at 6 MV beam quality, different phantoms were scanned and reconstructed. The agreement between a diamond detector and BIS responses is quantitative. Tomographic reconstruction of a complete set of these transmission profiles resulted in images which resolve 3 cm large objects having a (theoretical) contrast to water of less than 9%. Three millimetre objects with a 100% contrast are clearly visible. The BIS signal was shown to measure photon fluence distributions. The reconstructed images possess a spatial and contrast resolution sufficient for accurate imaging of the patient anatomy, needed for treatment verification in many clinical cases. PMID- 9869036 TI - Fourier transform Raman spectroscopy of polyacrylamide gels (PAGs) for radiation dosimetry. AB - Polyacrylamide gels (PAGs) are used for magnetic resonance imaging radiation dosimetry. Fourier transform (FT) Raman spectroscopy studies were undertaken to investigate cross-linking changes during the copolymerization of polyacrylamide gels in the spectral range of 200-3500 cm(-1). Vibrational bands of 1285 cm(-1) and 1256 cm(-1) were assigned to acrylamide and bis-acrylamide single CH2 deltaCH2 binding modes. Bands were found to decrease in amplitude with increasing absorbed radiation dose as a result of copolymerization. Principal component regression was performed on FT-Raman spectra of PAG samples irradiated to 50 Gy. Two components were found to be sufficient to account for 98.7% of the variance in the data. Cross validation was used to establish the absorbed radiation dose of an unknown PAG sample from the FT-Raman spectra. The calculated correlation coefficient between measured and predictive samples was 0.997 with a standard error of estimate of 0.976 and a standard error of prediction of 1.140. Results demonstrate the potential of FT-Raman spectroscopy for ionizing radiation dosimetry using polyacrylamide gels. PMID- 9869037 TI - Quantitative autoradiography using a radioimager based on a multiwire proportional chamber. AB - Determination of the biodistribution of radiopharmaceuticals is an important issue for the evaluation of their performance in diagnosis and therapy. In this study, we evaluated a digital radioimager (RI) based on a multiwire proportional chamber for quantitative autoradiography (AR). The RI allows direct detection of electronic emissions of gamma emitters. Its qualitative and quantitative performances were tested on 99mTc and (111)In labelled sections and compared with conventional film AR. Linearity of count rate versus activity was verified over a 104 range of activity. As compared with film AR, a substantial improvement of the detection limit was obtained even for acquisition periods up to 20 times less than film exposure times. We provided the basis for quantitative analysis with tissue equivalent paste standards: the 99mTc and (111)In RI counting efficiencies were respectively 1.19% and 2.35%. We illustrated the respective values of RI and film AR in two rat studies: 99mTc-DMSA in kidney and dual-isotope 99mTc-MIBI and (111)In-antimyosin in heart. Calculated activity concentrations on sections of rat organs confirmed good correlation to gamma counting (deviation less than 12%). We suggest RI as a convenient technique for fast localization of single or dual-isotope tracers and determination of activity distribution. PMID- 9869038 TI - Computer-aided analysis of conditions for optimizing practical electrorotation. AB - Previous studies have indicated that the variations in torque induced in particles in electrorotation electrode arrays are sufficiently large to cause errors in electrorotation measurements. In order to avoid this, experimenters usually study particles bounded by an arbitrary region near the centre of the electrodes. By simulating the time-dependent electric field for polynomial electrodes, we have assessed the variation in torque across the centre of the array. By considering both the variation in applied torque and the dielectrophoretic force in the electrode chamber, the optimal conditions for electrorotation experiments have been determined. Further to this, by comparing the torque variation across the electrode chamber for a number of common electrode designs, a comparison of the suitability of each electrode design for multiparticle electrorotation analysis has been made. PMID- 9869039 TI - Application of a video-optical beam imaging system for quality assurance of medical accelerators. AB - Method validation techniques were developed and experiments were carried out using a beam imaging system (BIS, Wellhofer Dosimetrie, Schwarzenbruck, Germany) for routine quality assurance of medical accelerators. The routine quality assurance tasks include x-ray beam flatness and symmetry check, light/radiation field congruence test, beam energy constancy for electrons and mechanical checks for couch and collimator rotations. Comparisons were made between the BIS application and conventional quality assurance methods that use radiographic films or detector arrays. In this work, we have demonstrated efficiency and accuracy of the BIS to perform some of the routine quality assurance tasks for medical linear accelerators. PMID- 9869040 TI - Fetal carotid blood flow during videofetoscopy. AB - BACKGROUND/PURPOSE: Intracranial bleeding has been reported as one of the complications of both open and minimally invasive fetal surgery and putatively attributed to intraoperative fluctuations of carotid blood flow. The aim of this study was to look at fetal carotid blood flow and its relationship with umbilical blood flow, blood pressure, oxygen delivery, and acid-base status in the fetus at various intraamniotic pressures with both liquid and gas media during fetoscopic surgery. METHODS: Six 115- to 130-day-gestation ewes underwent continuous invasive systemic blood pressure monitoring in the descending aorta. A hysterotomy was performed. A 6-mm ultrasonic blood flow probe was placed around the common umbilical artery at its origin from the fetal aorta. This was followed by placement of a double-lumen, 4F catheter in the fetal descending aorta through a femoral artery. A 4-mm ultrasonic blood flow probe was then placed around the fetal left common carotid artery. A pressure-monitoring, multiperforated catheter was placed inside the amniotic cavity. The fetus was repositioned inside the uterus, which was then closed. The abdominal wall was closed loosely. No further manipulation was performed for 1 hour. Intraamniotic pressure was raised from 0 to 30 mm Hg at 5-mm Hg intervals by infusing either warmed saline or medical air. Common umbilical artery and left carotid artery blood flows, blood pressure, blood gases, bicarbonate, sodium, and hematocrit were recorded in all fetuses at each 5-mm Hg interval. Maternal systemic blood pressure, O2 saturation, and temperature were kept constant. RESULTS: Carotid blood flow remained stable within the intra-amniotic pressure range studied (0 to 30 mm Hg), despite the significant drop in common umbilical artery blood flow uniformly observed above 20 mm Hg when saline was infused and above 15 mm Hg when air was infused. There was fetal hypoxemia and hypercarbia concomitant with decreased common umbilical artery blood flow (however, without fetal acidosis, because of compensatory elevation of bicarbonate). Within the intraamniotic pressure range studied, fetal aortic blood pressure, sodium, and hematocrit did not vary significantly, even when there was decreased umbilical blood flow. CONCLUSIONS: Fetal carotid blood flow is protected, possibly autoregulated, remaining stable even after umbilical blood flow decreases as a consequence of elevated intrauterine pressures up to 30 mm Hg during videofetoscopy. These data suggests that perioperative intracranial bleeding related to videofetoscopic surgery is caused by factors other than fluctuations of cerebral blood flow. PMID- 9869041 TI - Tracheal occlusion in the fetal rat: a new experimental model for the study of accelerated lung growth. AB - BACKGROUND: Prenatal tracheal occlusion accelerates fetal lung growth, but the mechanism of this phenomenon is unknown. Previous animal models have been limited by expense, lack of species-specific molecular probes, or the stage of lung development when studies could be performed. To provide a model that is more amenable to systematic analysis, we have developed an in vivo rat model of prenatal tracheal occlusion. METHODS: Time-dated pregnant rats underwent laparotomy at 19 days' gestational age (term, 22 days). The fetal head and neck were exteriorized through a hysterotomy, and the trachea was ligated under a dissecting microscope. The fetus was returned to the amniotic cavity, and the uterine and maternal abdominal incisions were closed. The dam and the fetuses were killed at 21.5 days' gestational age, and the fetal lungs were assessed for lung growth and compared with nonoperated littermate controls. RESULTS: Thirty two of 50 manipulated fetuses survived. Of the 32 survivors, successful tracheal ligation was confirmed in 20, and these 20 fetuses were compared with 33 littermate controls. Fetal body weight (4.81+/-0.26 g v 4.87+/-0.41 g) and heart weight (0.05+/-0.01 g v 0.05+/-0.01 g) were not significantly different between ligated fetuses and littermate controls, whereas the wet lung weight (0.30+/-0.06 g v 0.13+/-0.02 g, P<.01), lung-to-body-weight ratio (6.34+/-1.16% v 2.64+/ 0.41%, P<.01), dry lung weight (17.4+/-2.45 mg v 12.1+/-1.87 mg, P<.01), total lung DNA (1210+/-371 microg v 828+/-208 microg, P<.01), and total lung protein (14.3+/-5.3 mg v 8.7+/-1.7 mg, P<.01) were increased significantly in the ligated fetuses. The enlarged lung demonstrated normal histology findings after inflation fixation. CONCLUSIONS: Prenatal tracheal occlusion during the canalicular stage of lung development accelerates lung growth in the rat. In comparison with other large animal models, this relatively inexpensive small animal model has the distinct advantages of a short gestation, a large number of fetuses per litter, the availability of a developmental model of congenital diaphragmatic hernia, and the availability of well-defined molecular probes to investigate the mechanism of tracheal occlusion-induced lung growth. PMID- 9869042 TI - Thoracoscopic treatment of mediastinal cysts in children. AB - BACKGROUND/PURPOSE: The development of thoracoscopic surgery has made many procedures possible, including the treatment of mediastinal cysts in children. The authors report their experience with this procedure between 1992 and 1997. METHODS: Surgery was performed on 22 children aged from 1 month to 9 years (median, 27 months), weighing 5 to 49 kg (median, 12.5 kg). Diagnosis was made by antenatal ultrasound scan in six cases (27%), with a chest x-ray performed for respiratory symptoms in 14 cases, and with a chest x-ray performed for positive tuberculin intradermoreaction in two cases. Decision to resect the cyst was determined by thoracoscopy in 21 of the 22 cases, and by open surgery in one case only (subcarinal compressive cyst with left lung distension and a mediastinal shift). RESULTS: Eighteen of the 21 (86%) cases were treated successfully by thoracoscopy. In three cases of bronchogenic cysts, we performed an associated thoracotomy because the dissection was too difficult and dangerous. In three cases, a small part of a common wall between the cyst and the bronchus was not removed. The pathological diagnosis was bronchogenic cysts in 15 cases (71%), pleuropericardiat cysts in three cases (14%), esophageal duplication in two cases (10%), and cystic hygroma in one case (5%). Two postoperative complications were observed: one esophageal wound and a case of recurrent pneumothorax after chest tube removal. Patients were discharged after 2 to 11 days (median, 3 days). Follow-up was uneventful. CONCLUSIONS: Treatment of mediastinal cyst by thoracoscopy is feasible in most cases. Compressive cysts with lung distension and mediastinal shift remain a contraindication. If the cysts have a common wall with the bronchus or esophagus, or if they are subcarinal, the dissection may be difficult and dangerous, and thoracotomy may be preferable. PMID- 9869043 TI - The Edmonton experience with venovenous extracorporeal membrane oxygenation. AB - BACKGROUND/PURPOSE: Despite the proven effectiveness of venovenous extracorporeal membrane oxygenation (VV ECMO) in the treatment of neonates with severe respiratory failure, this technique is not widely used. The purpose of this study was to assess the authors' policy of preferred use of VV ECMO with a cephalad catheter and to compare the results with those of the Extracorporeal Life Support Organization (ELSO) Registry. METHODS: Charts of neonatal ECMO candidates were reviewed retrospectively. Data were collected for gestational age, birth weight, and diagnosis. Severity of illness was assessed by oxygenation index, lactate levels, and inotropic requirements before cannulation. Patients were divided into three groups: venovenous (VV), venoarterial (VA), and VV to VA ECMO. A cephalad catheter was inserted in the distal part of the jugular vein. RESULTS: Sixty-five neonates were supported with ECMO. Cannulation with a double lumen venovenous (VVDL) catheter was attempted in 63 neonates and successfully accomplished in 57. A survival rate of 86% was observed in neonates initially placed on VV ECMO. Five neonates initially placed on VV ECMO underwent conversion to VA ECMO. CONCLUSIONS: This study showed that the authors' preferred policy of VV ECMO did not result in an increase in mortality rate based on a comparison with ELSO data. VV ECMO with a cephalad catheter provides adequate support for unstable neonates with respiratory failure. PMID- 9869044 TI - Treatment of ornithine transcarbamylase deficiency in girls by auxiliary liver transplantation: conceptual changes in a living-donor program. AB - BACKGROUND/PURPOSE: Ornithine transcarbamylase (OTC) deficiency is an X chromosome-linked genetic disorder resulting in hyperammonemia hepatic dysfunction, coma, and serious neurological sequelae. This report describes an experience in treating this condition with living-related liver transplantation. METHODS: Three children with OTC were treated with a living-related liver transplantation. Potential donors were evaluated with the allopurinol loading test to exclude heterozygotes. FK506 and low-dose steroids were used for immunosuppression. Auxiliary partial orthotopic transplantation (APOLT) was used in two of the cases. RESULTS: All three children survived and are doing well without protein restriction. CONCLUSIONS: These observations suggest that in OTC removal of the native liver is not necessary because (1) a partial liver segment with normal enzyme activity corrects the hyperammonemia, (2) in case of graft failure the native liver is an available backup, and (3) the native liver has the ability to recover function. As long as living-related donors serve as the main source of donor organs, APOLT is a safer approach to liver transplantation in OTC. PMID- 9869045 TI - Intestinal anastomosis in children: a comparative study between two different techniques. AB - PURPOSE: The aim of this study was to compare the incidence of surgical complications between two different surgical techniques for intestinal anastomosis in children. METHODS: This was a clinically controlled, randomized study with blind follow-up from 18 to 36 months performed at the Reference Government Hospital in Mexico City. Eighty-six children required intestinal anastomosis, ages ranged between 1 month and 16 years, with emergency or elective surgery. Anastomoses of duodenum, rectum, with enteroplasty or protected with a proximal stoma were excluded. Two randomized groups were formed: (1) anastomosis with one layer of suture (Gambee stitches) and (2) with two layers of suture (first with Connel-Mayo stitches then with Lembert). Both groups were controlled in the principal variables without differences, and the follow-up concerning postoperative recovery was blind for the surgical team. RESULTS: Forty-two cases in group 1 and 44 in group 2 were compared. Intestinal dehiscence was found in 5 of 86 (5.8%), two from group I and three from group II (P value, not significant). Surgical time for anastomosis with one layer was an average of 26 minutes versus 43 minutes with two layers (P<.001). There were no stenoses within the follow-up period. CONCLUSIONS: This study proves that intestinal anastomosis with one layer of suture is as safe as anastomosis with two layers in children, and the time spent for completion of the procedure is significantly less with one plane of suture. For those reasons, it is the method of choice for intestinal anastomosis in children. PMID- 9869046 TI - Continence in patients who undergo posterior rectal flap anorectoplasty. AB - PURPOSE: The authors report the postoperative follow-up results of patients who underwent posterior rectal flap anorectoplasty (PRFA), with emphasis in fecal continence. Variables considered on each patient were sex, presence of fistula, location of the fistula, severity of the defect during the correction, sacral condition, dysraphism, and age at surgery. METHODS: Continence was rated using the Pena method and defecogram. The authors studied 20 patients (17 boys and three girls). RESULTS: Postoperatively, normal continence was found in 18 patients. Of the two remaining (boys), one had grade II spotting (sacrum agenesia and uretrobulbar fistula) and the other had grade II staining; this patient had the most severe malformation requiring an abdominoperineal approach. All of the defecograms showed complete emptying of the rectum. No recurrent fistulas occurred. CONCLUSIONS: The posterior rectal flap anorectoplasty is a new technique that takes into consideration all known factors that contribute to continence. This initial report confirms that this technique results in excellent continence for patients with imperforate anus. PMID- 9869047 TI - The role of cellular motility in the invasion of human neuroblastoma cells with or without N-myc amplification and expression. AB - BACKGROUND/PURPOSE: Patients who have neuroblastomas with N-myc amplification that are extremely invasive and result in distant metastases tend to have a very poor prognosis. The authors reported previously that N-mycamplification and expression might be closely related to the invasiveness of human neuroblastoma cells. However, the role of cellular motility has not yet been clarified in the invasion of neuroblastoma cells. The aim of this study was, therefore, to elucidate the role of cellular motility in the invasion of neuroblastoma cells. METHODS: Six human neuroblastoma cell lines were used for an invasion assay in vitro using polycarbonate filters coated with basement membrane Matrigel. The amplification and expression of N-myc oncogene was examined by Southern and Northern blotting, respectively. The cellular motility was quantified by computerized image analysis on the morphology of cultured cells. RESULTS: IMR-32, GOTO, and DZ, all of which had N-myc amplification, showed a high degree of invasiveness and a high cellular motility, whereas NB-69 and SK-N-SH without N myc amplification showed an extremely low degree of invasiveness and cellular motility. ST without N-myc amplification, which was established from an aggressive tumor, showed an exceptionally high degree of motility and invasiveness. A transcriptional reduction of the N-myc gene by retinoic acid (RA) decreased the motility, which thus resulted in a marked decline of invasiveness in IMR-32 and GOTO. CONCLUSION: The cellular motility correlated with the invasive capacity of human neuroblastoma cells, which thus indicated that cellular motility may play an important role in invasion. PMID- 9869048 TI - Transcatheter arterial chemoembolization in the treatment of hepatoblastoma. AB - BACKGROUND: The prognosis of hepatoblastoma is poor unless the tumor is completely resected. Various types of chemotherapy have been developed to increase its resectability. Recently, transcatheter arterial chemoembolization (TACE) has been developed for the treatment of unresectable adult hepatoma with favorable results. The authors applied this procedure to hepatoblastoma in infants and children. METHODS: TACE was performed in eight hepatoblastoma cases. After an intraarterial catheter was inserted into the main feeding artery of the tumor, injection of adriamycin or THP-adriamycin (20 to 30 mg/m2) dispersed in lipiodol and cisplatin (40 to 60 mg/m2) followed by embolization using Gelfoam pieces was performed. Effects of TACE were evaluated according to shrinkage of tumor mass on imaging examinations, alpha-fetoprotein (AFP) levels, and pathological findings of the surgical specimens 4 weeks after TACE. RESULTS: A marked reduction in tumor size associated with a decrease in AFP level occurred 1 month after the treatment. Tumor shrinkage ranged from 0.9% to 45.0% with a mean value of 25.8%. AFP levels decreased by 0.2% to 11.9% with a mean level of 4.6% from initial levels. In addition, there was no marked chemotherapeutic agent induced toxicity noted during the observation period. Resection of the tumors was performed safely after TACE in all cases. Pathological examination showed massive necrosis in the surgical specimens, and the mean percentage of necrotic area within the tumor was 71.1%. Two patients died of extensive lung metastasis 2 months and 3 years after the operation, respectively. The remaining six were doing well and free of disease at a mean follow-up period of 50 months. CONCLUSION: TACE is an effective, safe, and useful method for the initial treatment of hepatoblastoma. PMID- 9869049 TI - The effects of prenatal intraamniotic surfactant or dexamethasone administration on lung development are comparable to changes induced by tracheal ligation in an animal model of congenital diaphragmatic hernia: studies of lung glycogen content, elastic fiber density, and collagen content. AB - BACKGROUND/PURPOSE: A new noninvasive therapeutic strategy, which consisted of prenatal intraamniotic administration of porcine surfactant or dexamethasone, was previously used to prevent the functional and structural immaturity of lungs associated with congenital diaphragmatic hernia (CDH), and its effects on lung development were comparable with the changes induced by tracheal ligation (TL). The purpose of this study is to verify if this novel therapeutic modality has any effect in the elevated concentration of lung glycogen and altered contents of lung elastic fiber and collagen promoted by CDH. METHODS: A pilot study was performed to investigate in the rabbit model if the infused drugs in the amniotic cavity were aspirated by the CDH and non-CDH fetuses, and if there was correspondence between lung immaturity and high glycogen concentration in lung tissue. Experimental groups consisted of 50 pregnant rabbits that underwent surgery on gestational day 24 or 25 to create left-sided diaphragmatic hernias in 56 fetuses, which were divided in groups according to the procedures: CDH (n = 12), CDH plus TL (n = 16), CDH plus intraamniotic administration of Curosurf (40 mg, n = 12), and CDH plus intraamniotic administration of dexamethasone (n = 16). On gestational day 30, the fetuses were delivered by cesarean section, and 28 normal unoperated fetuses served as controls. The lungs were weighed and submitted to biochemical determination of glycogen, morphometric evaluation of elastic fibers, and colorimetric analysis of collagen. RESULTS: In all CDH and non-CDH fetuses of the pilot study, the amniotic content was massively aspirated into the lungs and trachea. There was an increase in lung glycogen content of fetuses at 24 days' gestation in comparison with 20-day gestational age fetuses, followed by a decrease in the near full-term fetuses. In the fetuses of the experimental groups, CDH decreased the lung weight to body weight ratios of lungs ipsilateral to the hernia. These changes were reversed by TL but not by intraamniotic administration of surfactant or dexamethasone. Lung glycogen concentrations in the lungs of CDH fetuses were significantly higher than those in the control group. These changes were reversed by intraamniotic administration of surfactant but not by dexamethasone administration or TL. In the lungs ipsilateral to the hernia, surfactant administration promoted a significant decrease in glycogen content to levels lower than control lungs. CDH promoted a decrease in the linear density of elastic fibers in both lungs, ipsilateral and contralateral to the hernia. This alteration was partially corrected by TL and surfactant administration, although dexamethasone administration had no effect. The concentrations of collagen in both lungs were increased significantly by CDH, and these alterations could not be reversed by TL. In the lungs ipsilateral to the hernia, intraamniotic administration of surfactant or dexamethasone promoted a significant decrease in the lung concentration of collagen but not to control levels. CONCLUSIONS: The positive effects of intraamniotic surfactant or dexamethasone administration on lung maturity of fetuses with CDH were observed. This therapy may be a substitute for TL. PMID- 9869050 TI - Ultrasonography for inguinal hernias in boys. AB - BACKGROUND/PURPOSE: History taking and physical examination alone no longer meet the surgeon's need in the diagnosis of inguinal hernia. Ultrasonography (US) provides a good and safe diagnostic tool for inguinal hernias in boys. METHODS: From 1995 to 1997, 244 boys with inguinal hernias (41 bilateral and 203 unilateral), received preoperative US on both groins to confirm the diagnosis. Those with positive US findings, such as viscera or fluid in inguinal canal or widening of the internal inguinal ring, underwent surgery. RESULTS: The accuracy of diagnosis with US and clinical assessment were 97.9% and 84%, respectively. More than 95% of widening of internal inguinal rings (>4 mm) proved to be hernias. There were two direct inguinal hernias and two femoral hernias, which were misdiagnosed by clinical examination, but proved to be diagnosed correctly by US. CONCLUSIONS: US serves as a noninvasive and highly accurate diagnostic tool for inguinal hernias in boys. Using 4 mm as the upper limit of the normal diameter of the internal inguinal ring, an occult inguinal hernia can be easily detected before surgery. PMID- 9869051 TI - Pedicled external spermatic fascia flap for urethroplasty in hypospadias and closure of urethrocutaneous fistula. AB - PURPOSE: To prevent the development of urethrocutaneous fistula after urethroplasty for hypospadias or recurrence after closure of a urethrocutaneous fistula, the authors developed a new simple technique wherein the readily available external spermatic fascia (ESF) surrounding the testis and cord is used as a pedicled blanket flap to cover the neourethra or the site of closure of a urethrocutaneous fistula. RESULTS: In three patients who had urethroplasty for hypospadias incorporating our ESF flap procedure, no urethrocutaneous fistula developed. It was also effective for closure of urethrocutaneous fistula in five patients, some of whom had had recurrent fistula formation. PMID- 9869052 TI - Testicular suture: a significant risk factor for infertility among formerly cryptorchid men. AB - BACKGROUND/PURPOSE: Although fertility is decreased after cryptorchidism, the importance of risk factors, including parenchymal testicular suture, is unknown. The aim of this study was to examine the relationship between parenchymal testicular suture and failure to conceive a child for 1 year or longer among formerly cryptorchid men. METHODS: Men who underwent orchidopexy between 1955 and 1972 at the Children's Hospital of Pittsburgh (n = 619) were surveyed by questionnaire and their medical records reviewed. Only the men who attempted to conceive a child (n = 387) are included. RESULTS: Logistic regression analysis determined significant risk factors for infertility. Testicular suture was strongly related to infertility (RR, 7.56; 95% CI, 1.66, 34.39) as were bilateral cryptorchidism (RR, 5.51; 95% CI, 1.58, 19.24), varicocele (RR, 4.72; 95% CI, 1.42, 15.75), hormone treatment before surgery (RR, 3.69; 95% CI, 1.22, 11.11), and partner conception problem (RR, 3.32; 95% CI, 1.11, 9.90). CONCLUSIONS: Testicular suture was a potent independent determinant of infertility among formerly cryptorchid men who have orchidopexy. Bilateral cryptorchidism, hormone treatment, varicocele, and partner conception problems also were associated with increased infertility. PMID- 9869053 TI - Characteristics of infantile cases of congenital dilatation of the bile duct. AB - PURPOSE: This study was performed to clarify whether infants with congenital dilatation of the bile duct (CDBD) have any specific characteristics in comparison with older children. METHODS: Of 153 cases with CDBD treated at our institution between 1959 and 1997, 34 (22.2%) were infants less than 1 year of age. The control group consisted of 46 children between 1 and 4 years of age. Symptoms, laboratory data, and histological findings were compared between these two groups. RESULTS: As the initial symptoms, abdominal pain was not present in infantile cases, with a significant difference from the older children, and a significantly higher incidence of abdominal mass and acholic stools was noted in infantile cases. Laboratory data of serum aspartate transaminase and amylase level and amylase concentration in the bile of the common bile duct were significantly lower in the infant group than in the older children. The ratio of morphological type of cystic to cylindrical dilatation was 33 to 1 in infants with a significant difference from the control group. CONCLUSION: Infantile cases of CDBD, which mostly have cystic type dilatation, in contrast to older children, are likely to present with abdominal mass. Pancreatitis with abdominal pain related to reflux of pancreatic juice into the biliary duct appears to be uncommon in infantile cases. PMID- 9869054 TI - Aesthetics and lateral thoracotomy in the neonate. AB - BACKGROUND: Lateral thoracotomy is a relatively common procedure in the neonate. Early reports on the incidence of shoulder deformity, scoliosis, and winged scapula rapidly led to a muscle-sparing approach. Scar cosmesis, however, received less attention. Better education and increasing sophistication have led to a higher aesthetic expectation by the public. METHODS: Since 1988 the authors have combined both principles in the high axillary skin crease approach to lateral thoracotomy in 27 children with esophageal atresia and tracheoesophageal fistula and two children with a patent ductus arteriosus. RESULTS: Access was not restricted, and early healing was uncomplicated by wound breakdown or infection. Scar aesthetics is excellent and is enhanced by the natural tendency of the scar to migrate into the axilla. Long-term morbidity has been singularly absent. CONCLUSION: Parent appreciation and operator satisfaction have been noteworthy. PMID- 9869055 TI - Nissen fundoplication may induce gastric myoelectrical disturbance in children. AB - BACKGROUND: Recurrent vomiting with failure to thrive is a common problem in neurologically impaired children. Many undergo fundoplication to control the underlying gastroesophageal reflux, but the results of surgery are not always satisfactory, and postoperative retching may be a major problem. Retching is part of the emetic reflex and is associated with nausea, which is itself associated with disturbed gastric electrical control activity, resulting in a gastric dysrhythmia. METHODS: By recording gastric electrical control activity before and after Nissen fundoplication using the noninvasive technique of surface electrogastrography, the authors have shown that (1) Neurologically impaired children with gastroesophageal reflux more commonly have a preexisting gastric dysrhythmia (65% neurologically impaired v 20% neurologically normal children with gastroesophageal reflux, P<.05), (2) Children who retch preoperatively are three times more likely to retch postoperatively, and (3) 25% of neurologically impaired children may start to retch postoperatively for the first time. CONCLUSION: The authors propose that in neurologically impaired children, loss of central inhibitory mechanisms may result in inappropriate activation of the emetic reflex, which may be heightened by antireflux surgery. PMID- 9869056 TI - Intraabdominal pyloromyotomy through the umbilical route: a technical improvement. AB - BACKGROUND/PURPOSE: Performing a pyloromyotomy through a supraumbilical skin fold incision will leave an almost invisible scar and therefore has definitive cosmetic advantages. This alternative approach may be related to technical difficulties in delivering a large pyloric tumor when compared with the conventional pyloromyotomy through a right upper quadrant incision. However, in situ (intraabdominal) myotomy can help overcome this inconvenience. METHODS: Of 122 cases of infantile hypertrophic pyloric stenosis operated on between January 1990 and August 1996, 29 underwent a pyloromyotomy performed intraabdominally through the umbilical route. The medical records of these babies were reviewed. RESULTS: Twenty-three boys and six girls (median age, 30 days; range, 17 to 70 days) underwent surgery. The median hospital stay was 2.5 days. There were two intraoperative technical complications (small mucosal perforation) and one postoperative wound complication (abcess formation) requiring local drainage. CONCLUSIONS: In situ pyloromyotomy through the umbilical route is an elegant alternative in cases of a large pyloric tumor. PMID- 9869057 TI - Neurovascular morbidity from the lithotomy position. AB - The lithotomy position is used in pediatric surgery when it is necessary to have simultaneous access to the abdomen and perineum. At the authors' institution, after a prolonged period in the lithotomy position, significant lower limb complications developed in four patients. Two patients with Hirschsprung's disease underwent a redo Duhamel procedure, one had an anorectal leiomyoma excised, and one had an ileoanal anastomosis for ulcerative colitis. Sciatic nerve injury developed in two patients and deep venous thrombosis and bilateral compartment syndrome resulting in myonecrosis developed in one each. There is an association between operations that require prolonged lithotomy position and the development of postoperative neurovascular complications. By placing the child in the lithotomy position only when access to the perineum is required, these significant injuries may be avoided. PMID- 9869058 TI - Dermatofibrosarcoma protuberans and the Bednar tumor: treatment in the pediatric population. AB - Dermatofibrosarcoma protuberans (DFSP) is a low-intermediate-grade cutaneous sarcoma that has a marked propensity for local recurrence after excision. The Bednar variant of this tumor is even less common and is distinguished histologically by the dispersal of melanin containing cells in an otherwise typical DFSP. Both are considered to be tumors of the third and forth decades of life, but both DFSP and the Bednar variant have been described in children. Until this report of a congenital Bednar tumor, only the DFSP has also been described in the neonate. The histopathology and surgical management of DFSP and Bednar tumors are outlined with emphasis on reported experience in the pediatric population. The surgical management of these lesions in children is based on numerous series in adults and the limited pediatric experience. The recommended treatment is wide excision with 3-cm margins of visibly uninvolved tissue and inclusion of superficial fascia. PMID- 9869059 TI - Gastric duplication-colonic fistula with colonic ulceration and bleeding. AB - An 11-month-old girl had massive rectal bleeding. A midabdominal mass was palpated. Ultrasound scan showed a large cystic mass with diffusely echogenic contents; Tc-99M radionuclide scan confirmed a duplication. Exploratory surgery and pathology examination showed a gastric duplication with formation of a fistula into the transverse colon with a colonic ulcer and hemorrhage. This combination of findings is presented as a rare cause of lower gastrointestinal hemorrhage in infancy. PMID- 9869060 TI - Lung abscess: an unusual presentation of congenital tracheoesophageal fistula without atresia. AB - Tracheoesophageal fistula (TEF) without atresia is rare and usually presents with symptoms from birth. In this report, a 9-year-old boy presented with productive cough of 4 month's duration and was shown to have a right lung abscess seen on chest radiograph. His parents denied earlier respiratory symptoms or illnesses. Rigid bronchoscopy showed a fistulous opening of about 1 mm in diameter in the posterior wall of the trachea about 16 cm from the upper incisor teeth. Cannulation with a ureteral catheter demonstrated that the fistulous opening communicated with the esophageal lumen. The tracheoesophageal fistula was 1 cm long and was divided through a right supraclavicular incision. The postoperative period was uneventful, and the patient was discharged on the third postoperative day. This case demonstrated that TEF should be considered in any patient presenting with chronic respiratory problems even after a prolonged symptom-free period. PMID- 9869061 TI - Mesenteric venous thrombosis after appendicectomy in a child: clinical case and review of the literature. AB - Mesenteric venous thrombosis (MVT) after appendicectomy has not been reported in the pediatric literature. This complication is associated with significant morbidity and mortality rates in adults. The report of this case attempts to analyze the features found in an appendicectomized child documented by radiological examination and surgery before and after starting heparin therapy. Anticoagulation treatment should be started as soon as possible and continued for a long time to decrease the risk of relapse. PMID- 9869062 TI - Cardiac tamponade: a rare but life-threatening complication of central venous catheters in children. AB - The widespread use of central venous catheters in the treatment of pediatric patients has caused an increased incidence of complications. A rare, but potentially fatal complication occurs when the heart is perforated by the catheter tip causing a cardiac tamponade. This perforation of the heart generally is associated with the insertion procedure, but may also occur after some time because of displacement of the catheter tip. The authors present three cases in which the placement of a central venous catheter resulted in lethal cardiac tamponade. Proper positioning of the catheter tip in the superior vena cava and a high index of suspicion are essential in preventing this serious complication. Contrast-enhanced chest x-ray after insertion of the catheter must be performed to ascertain a correct position of the tip. PMID- 9869063 TI - Prune belly syndrome with penile and urethral agenesis: report of a case. AB - The authors report the case of an infant born with prune belly syndrome associated with penile and urethral agenesis. At 15 weeks' gestation, antenatal ultrasonography showed a fetal giant bladder, congenital hydronephrosis, and oligohydramnios, and at 17 weeks' gestation, a fetal vesicoamniotic shunt operation was performed. A boy was born at 33 weeks' gestation with prune belly syndrome, an anocutaneous fistula, and penile agenesis. A cystostomy and cut-back operation were performed immediately, showing urethral agenesis, no urethral opening, and left renal hypoplasia. Thereafter, his renal system began functioning normally, and a urinary tract infection resolved. The authors speculate that the prune belly syndrome in this patient was caused by penile and urethral agenesis. PMID- 9869064 TI - Azygous lobe presenting as a posterior mediastinal mass in a 2-year-old boy. AB - Posterior mediastinal masses encompass a varied group of surgical lesions that include malignant diseases as well as the anatomic malformations of four organ systems. The authors report on a 2-year-old child with a posterior mediastinal mass who was found at thoracotomy to have a chronically infected "hepatized" azygous lobe of the right lung. The authors review current literature on this topic as well as the embryogenesis of azygous lobes. PMID- 9869065 TI - Pseudoexstrophy associated with congenital pouch colon. AB - Pseudoexstrophy or covered exstrophy is a rare exstrophy variant. The authors report a case of covered exstrophy that presented as a newborn with widely separated pubic bones and rectus muscles, a low-set umbilicus, and a subcutaneous bladder. The anal opening was absent, and there was a complex malformation of the external genitalia consisting of a small, laterally displaced penis and a right sided ectopic hemiscrotum. Micturition and urinary continence were normal. The child also had a high anorectal malformation with a coexistent type IV congenital pouch colon (CPC) malformation. Both kidneys were normal. Preliminary surgery consisted of a divided sigmoid colostomy proximal to the colonic pouch. The literature is reviewed and the embryogenesis of pseudoexstrophy and its associated malformations are discussed. PMID- 9869066 TI - Acute scrotum due to edidymo-orchitis associated with vasal anomalies in children with anorectal malformations. AB - Epididymo-orchitis, an uncommon cause of acute scrotum in prepubertal boys, is infection or inflammation of epididymis and testis. Epididymo-orchitis may be associated with urinary tract infections or reflux of urine predisposed by an underlying vasal anomaly. Two infants with anorectal malformations who presented with acute scrotum are reported. The surgical exploration of the testes showed findings consistent with epididymo-orchitis. Further radiological investigations of urinary tract showed vasal anomalies in both patients. If a patient with anorectal malformation presents with acute scrotum, epididymo-orchitis should be suspected initially. Evaluations should be directed toward defining predisposing vasal anomaly, and appropriate therapeutic measures should be undertaken to prevent recurrences. PMID- 9869067 TI - Familial occurrence of intestinal obstruction in children with the syndrome of mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS). AB - The syndrome of mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) is an uncommon neuromuscular disorder caused by mitochondrial dysfunctions that result in headaches, seizures, and progressive dementia. The authors describe a clinical case study of gastrointestinal manifestations in a pedigree with MELAS, in which all three children, ages 11, 8, and 6, demonstrated acute onset of intestinal obstruction. They unexpectedly showed severe abdominal distension and vomiting. Their parents had no clinical manifestation. The first female sibling underwent an emergent laparotomy because she was diagnosed to have intestinal strangulation. She had postoperative complications caused by progressive lactic acidosis and died the next day. The second and third sisters had similar onsets of the disease and were treated with gastrointestinal decompression and intravenous administration of lactate-free fluid and coenzyme Q10. Genetic testing using blood samples showed an A-to-G point mutation at nucleotide position 3243 in the tRNALeu(UUR) region in the mitochondrial DNA. In MELAS children who demonstrate acute onset of gastrointestinal manifestations, a careful review of family history and an elevation of serum lactate and pyruvate levels may enable a differential diagnosis to be made of acute abdomen to avoid unnecessary surgical intervention. PMID- 9869068 TI - Experience with vascular control before excision of giant, highly vascular sacrococcygeal teratomas in neonates. AB - Sacrococcygeal teratomas (SCT) are the most common neoplasms in newborns with a reported occurrence of 1 in 35,000 live births. Highly vascularized tumors in which the ratio of tumor weight to patient weight approaches 1:1 are frequently associated with hyperdynamic states, prenatal hydrops, placentamegaly, postnatal high-output cardiac failure, and carry a high perinatal mortality rate. Operative management of giant, highly-vascular sacrococcygeal teratomas in neonates can be complicated by life-threatening hemorrhage. Laparotomy, control of the aorta, and the arterial blood supply to these tumors before resection has been advocated as a safer alternative. The authors report their experience with three infants successfully treated using this approach. Hemodynamic stability was maintained during the operations. All tumors were resected successfully. The patients are alive without evidence of recurrence, 8.5 months to 18 months (average, 14.3 months) after the operation. Alpha-fetoprotein levels dropped to normal range after the removal of SCT and have remained normal in follow-up. Vascular control before excision of giant, highly-vascular SCT in neonates is safe, decreases intraoperative blood loss, and postoperative morbidity. PMID- 9869069 TI - Inflammatory pseudotumor causing intestinal obstruction: diagnostic and therapeutic aspects. AB - The authors report an unusual presentation of inflammatory pseudotumor (IPT) that caused intestinal obstruction in a 9-year-old boy, and discuss the clinicopathologic features of this rare entity with emphasis on diagnosis and treatment. There are no specific presumptive clinical and laboratory findings, including tumor markers and imaging techniques, that distinguish mesenteric IPT from other abdominal mass lesions. The most important diagnostic aid is to bear this entity in mind when a child presents with intestinal obstruction associated with an abdominal mass. Radical unnecessary surgical procedures or potentially harmful therapy should be avoided, and appropriate treatment is achieved by total excision of the lesion in most of the cases. PMID- 9869070 TI - Perianal abscess and fistula-in-ano in infants. PMID- 9869071 TI - Left heart hypoplasia in neonates having congenital diaphragmatic hernia and treated with ECMO. PMID- 9869072 TI - Treatment of Hirschsprung's disease. PMID- 9869073 TI - Preterm C-section delivery for fetal gastroschisis. PMID- 9869074 TI - Hypothermia-related deaths--Georgia, January 1996-December 1997, and United States, 1979-1995. AB - Although hypothermia-related deaths are prevalent during the winter in states that have moderately cold (e.g., Illinois, New York, and Pennsylvania) to severely cold (e.g., Alaska and North Dakota) winters and in states with mountainous or desert terrain (e.g., Arizona, Montana, and New Mexico), hypothermia-related deaths also occur in states with milder climates (e.g., Georgia, Mississippi, and South Carolina), where weather systems can cause rapid changes in temperature. This report summarizes three hypothermia-related deaths in Fulton County, Georgia, representing persons in the highest risk groups for hypothermia; and summarizes hypothermia-related deaths in Georgia during January 1996-December 1997 and in the United States during 1979-1995. PMID- 9869075 TI - Lead poisoning associated with imported candy and powdered food coloring- California and Michigan. AB - Although the most common source of pediatric lead poisoning is dust within the home that contains deteriorated lead-based paint from walls and windowsills, other less common sources (1-3) can result in excess exposure among children (i.e., blood lead levels [BLLs] > or =10 microg/dL). This report describes two cases of pediatric lead poisoning associated with eating imported candy and food stuffs and underscores the importance of thorough history-taking to identify unusual sources of lead exposure. PMID- 9869076 TI - Update: respiratory syncytial virus activity--United States, 1997-98 season. AB - Respiratory syncytial virus (RSV) is the single most important cause of serious lower respiratory tract disease in infants and young children worldwide. In temperate climates, infections primarily occur during yearly outbreaks that usually peak during the winter months. RSV activity in the United States is monitored by the National Respiratory and Enteric Virus Surveillance System (NREVSS), a voluntary, laboratory-based system. This report summarizes trends for RSV reported to NREVSS from July 1997 to June 1998 and presents preliminary surveillance data from July 1 to November 18, 1998. PMID- 9869077 TI - Laboratory performance evaluation of N95 filtering facepiece respirators, 1996. AB - In 1995, CDC's National Institute for Occupational Safety and Health (NIOSH) introduced a new classification scheme for particulate air-purifying respirators. Most health-care workers use type N95 half-mask filtering facepiece respirators (i.e., N95 respirators) to prevent occupational transmission of tuberculosis. As a result, NIOSH received inquiries about how well N95 respirators fit, whether they need to be fit tested, and whether they can be quantitatively fit tested. In response to these inquiries, NIOSH evaluated the performance of 21 N95 respirator models on a 25-person panel. This report summarizes the results of this evaluation, which indicate that fit testing is needed to ensure at least the expected level of protection (i.e., the concentration of airborne contaminants inside the respirator is < or =10% of ambient levels). PMID- 9869078 TI - Progress toward global measles control and regional elimination, 1990-1997. AB - In 1989, the World Health Assembly resolved to reduce measles morbidity and mortality by 90% and 95%, respectively, by 1995, compared with disease burden during the prevaccine era. In 1990, the World Summit for Children adopted a goal of vaccinating 90% of children against measles by 2000. Regional measles elimination goals have been established in the American Region (AMR) by 2000, the European Region (EUR) by 2007, and the Eastern Mediterranean Region (EMR) by 2010. This report updates progress toward global measles control and regional elimination, and presents measles vaccination coverage and incidence for 1997 and WHO estimates of global measles morbidity and mortality in 1997 compared with the prevaccine era. PMID- 9869079 TI - Expression and functional analysis of glycosyl-phosphatidyl inositol-linked CD46 in transgenic mice. AB - BACKGROUND: Complement activation plays a pivotal role in hyperacute xenograft rejection. In humans, activation of complement is regulated by a number of cell surface regulatory proteins. Membrane cofactor protein (CD46) is one such regulator that protects cells by acting as a cofactor for the factor I-mediated cleavage of C3b and C4b. Transgenic animals expressing human CD46 may provide organs that are resistant to complement attack. However, attempts to generate mice expressing human CD46 using cDNA-based constructs have been largely unsuccessful. METHODS: Transgenic mice expressing a glycosylphosphatidyl inositol (GPI)-linked form of CD46 were generated by microinjection of a hybrid CD46/CD55 cDNA under the control of the human intercellular adhesion molecule-2 promoter. Expression of CD46-GPI on the vascular endothelium was determined by immunohistochemistry. The ability of CD46-GPI to protect mouse tissues from human complement attack was determined using an ex vivo isolated perfused heart model. RESULTS: Three founder animals expressing CD46-GPI were identified. Histological analysis showed strong and uniform expression of CD46-GPI on the vascular endothelium of all organs examined. Ex vivo perfusion of transgenic mouse hearts with human plasma showed a reduction in C3c deposition and a slightly prolonged function compared with controls. CONCLUSIONS: High-level expression of CD46-GPI was achieved in transgenic mice by using a modified cDNA-based construct. The CD46-GPI was functional, providing some protection from complement-mediated damage in the ex vivo model, and may be useful in xenotransplantation if expressed in combination with CD55 and CD59. PMID- 9869080 TI - Evidence that macrophages are required for T-cell infiltration and rejection of fetal pig pancreas xenografts in nonobese diabetic mice. AB - BACKGROUND: Host macrophages are abundant within fetal pig pancreas xenografts undergoing rejection, but their role is unknown. Therefore, we examined the effect of host macrophage depletion on xenograft rejection. METHODS: Nonobese diabetic (NOD) mice were given clodronate-loaded liposomes intravenously to deplete macrophages. Controls received phosphate-buffered saline (PBS) or PBS liposomes. General immune status was assessed after 2, 3, and 7 days by (1) fluorescence-activated cell sorter analysis of peripheral blood, spleen, and lymph node cells, (2) immunohistochemistry on spleens, and (3) mixed lymphocyte reaction. Organ-cultured fetal pig pancreas was transplanted under the kidney capsule of NOD mice 3 days after clodronate or PBS injection. Grafts were assessed histologically at 4, 5, 6, and 8 days after transplantation. RESULTS: Splenic macrophages and peripheral blood monocytes were depleted 2 days after clodronate treatment but had recovered within 11 days. T cell, B cell, and dendritic cell numbers were normal in spleen, peripheral blood, and lymph nodes of clodronate-treated mice, and T cells and antigen-presenting cells from these mice functioned normally in mixed lymphocyte reaction. Clodronate treatment markedly reduced graft infiltration by macrophages, T cells, and eosinophils at 4, 5, and 6 days after transplantation, and was associated with maintenance of endocrine cell viability and insulin expression. However, all grafts were rejected 8 days after transplantation, concordant with reappearance of splenic macrophages. CONCLUSIONS: Short-term, specific depletion of macrophages markedly delayed cellular infiltration and rejection of xenografts. The results provide the first evidence that macrophages promote T-cell infiltration and rejection of fetal pig pancreas xenografts in NOD mice. PMID- 9869081 TI - Prolonged survival of rat liver allografts transfected with Fas ligand-expressing plasmid. AB - BACKGROUND: Transplantation of Fas ligand (FasL) gene-transfected tissues can have opposite effects. For example, cotransplantation of pancreas islets with myoblasts transfected with FasL-expressing plasmid vector (pFasL) prevented graft rejection, whereas the expression of FasL directly within islets using adenovirus vector led to graft destruction. It was also reported that FasL expression on pancreas islets led to neutrophilic infiltration and rapid destruction of the islets. From these results, overexpression of FasL in transfected tissues may lead directly to self destruction through an autocrine Fas-FasL pathway or graft destruction through neutrophil recruitment. To date there have been no reports of successful transplantation of FasL gene-transfected solid organs. METHODS: Rat pFasL was transfected at a dose of 90, 180, 270, or 360 microg into rat liver with an inactivated hemagglutinating virus of Japan conjugated to liposome vesicles (HVJ-liposome), and the gene-transfected livers were transplanted to allogeneic rats. RESULTS: In 18 rats transfected with 180 microg of pFasL, 14 (78%) did not develop fulminant hepatitis. FasL-mRNA was detected in these livers at 3, 5, 7, and 14 days after transfection. The expression of FasL protein was also observed in the transfected liver, and the transfection rate by this method was 11.1+/-1.9%. The livers were then transplanted to allogeneic recipients, resulting in significant (P<0.01) prolonged recipient survival times. Histological observation showed that the pFasL-transfected liver allografts caused apoptotic cell death in infiltrating activated T cells. In contrast, transfection of pFasL higher than 180 microg resulted in lethal hepatitis in all rats, and its low dose (90 microg) did not induce the hepatitis or prolong recipient survival. CONCLUSION: Our results indicate that rat liver allografts can be protected to host immune responses by an adequate level (approximately 10%) of FasL expression in the livers using HVJ-liposome incorporating pFasL. PMID- 9869082 TI - Estradiol inhibits allograft-inducible major histocompatibility complex class II antigen expression and transplant arteriosclerosis in the absence of immunosuppression. AB - BACKGROUND: The etiology of transplant arteriosclerosis is unknown, but current data point to the alloimmune response. Previously, we found that estradiol-17beta (E2) with immunosuppressant cyclosporine abolishes major histocompatibility complex (MHC) class II expression in the allograft. This study determines the effect of E2 on MHC class II antigen expression in the allograft, in the absence of immunosuppression. METHODS: Lewis male rats received orthotopic abdominal aorta allografts from male Brown-Norway rats. The recipients were treated continuously subcutaneously with either 20 microg x kg(-1) x day1 of E2 (n=20) or placebo (n=20), from 2 days before transplantation until death on posttransplant days 1, 3, 7, and 14. The allografts were harvested and processed for morphometry and for immunohistochemical staining of MHC class II antigens, macrophages, CD4 and CD8 T lymphocytes, interferon-gamma (IFN-gamma), and IFN-gamma receptor. RESULTS: With E2 treatment, we observed that inducible MHC class II antigen expression is abolished in the media of the vascular allograft; the expression of IFN-gamma and IFN-gamma receptor is unaffected; and macrophage infiltration of the vascular allograft is inhibited significantly (P<0.01), whereas the CD4 and CD8 T lymphocytes are not significantly (P=0.07) suppressed. The myointimal hyperplasia in the allografts from E2-treated-recipients was 3-4-fold less than that from the placebo-treated recipients. CONCLUSIONS: Without immunosuppression, E2 inhibition of transplant arteriosclerosis is still associated with inhibition of inducible MHC class II antigen expression in the allografts. The estradiol 17beta abolition of inducible MHC class II antigen expression in the aorta allograft occurs in spite of up-regulation of IFN-gamma ligand and receptor protein. PMID- 9869083 TI - Ethanol, not fat accumulation per se, increases free radical production in a low flow, reflow liver perfusion model. AB - BACKGROUND: Ethanol increases primary graft failure after liver transplantation, yet whether it acts via mechanisms involving fat accumulation remains unclear. METHODS: Rats were pair-fed a modified Lieber-DeCarli liquid diet containing 35% (high-fat) or 12% (low-fat) of calories as fat combined with 36% of calories as ethanol or isocaloric maltose-dextrin for 4-5 weeks. Reperfusion injury to the liver was studied using a low-flow, reflow perfusion model and a liver transplantation model, and free radicals were detected using electron spin resonance and the spin trapping technique. RESULTS: As expected, basal hepatic triglycerides were similar in livers from rats fed low- and high-fat control diets. Ethanol did not alter triglyceride levels significantly in rats fed a low fat diet, but increased values about 2.4-fold in rats fed a high-fat diet. Ethanol increased lactate dehydrogenase release during reperfusion from 10 to 26 IU/g/h in rats fed a low-fat diet and from 17 to 34 IU/g/h in rats fed a high-fat diet, respectively. Portal pressure increased from about 3 to 10.5 cm H2O upon reperfusion in livers from high-fat, ethanol-fed rats, but only reached values of 9.1 in the low-fat, ethanol-fed group. A free radical adduct signal was detected in the bile of livers from ethanol-treated rats, and the magnitude of this signal was similar in livers of ethanol-treated rats fed high- or low-fat diets. However, radical adducts could not be detected in either group in the absence of dietary ethanol. Moreover, 67-77% rats given low-fat or high-fat control diets survived after liver transplantation, but only 11% survived if treated with ethanol. CONCLUSIONS: It is concluded that ethanol plays a major role in hepatic reperfusion injury, most likely via mechanisms involving free radicals. Increased hepatic fat content alone plays only a minor role, probably by causing slight disturbances in the hepatic microcirculation. PMID- 9869084 TI - Disseminated intravascular coagulation in association with the delayed rejection of pig-to-baboon renal xenografts. AB - BACKGROUND: Intravascular fibrin deposition and platelet sequestration occur with porcine xenograft rejection by baboons. Disseminated intravascular coagulopathy may arise either as a direct consequence of the failure to fully deplete xenoreactive natural antibodies and block complement, or because of putative cross-species molecular incompatibilities in this discordant species combination. METHODS: Three baboons were conditioned with retrovirally transduced autologous bone marrow to induce tolerance to swine antigens. Xenoreactive natural antibodies and complement were depleted by plasmapheresis and the use of Gal alpha1-3Gal column adsorptions; baboons were then splenectomized and underwent renal xenografting from inbred, miniature pigs. Soluble complement receptor type 1 with protocol immunosuppression (mycophenolate mofetil, 15-deoxyspergualin, steroids, and cyclosporine) was administered. RESULTS: A bleeding diathesis was clinically evident from days 5 to 12 after transplantation in two baboons. Low levels of circulating C3a, C3d, and iC3b were measured despite the absence of functional circulating complement components. Profound thrombocytopenia with abnormalities in keeping with disseminated intravascular coagulopathy were observed. Prolongation of prothrombin and partial thromboplastin times was accompanied by evidence for tissue factor-mediated coagulation pathways, high levels of thrombin generation (prothrombin fragment F(1+2) production and thrombin-antithrombin complex formation), fibrinogen depletion, and production of high levels of the fibrin degradation product D-dimer. Importantly, these disturbances resolved rapidly after the excision of the rejected xenografts in two surviving animals. Histopathological examination of the rejected xenografts confirmed vascular injury, fibrin deposition, platelet deposition, and localized complement activation. CONCLUSIONS: Systemic coagulation disturbances are associated with delayed xenograft rejection. PMID- 9869085 TI - Excellent recovery after prolonged heart storage by preservation with coronary oxygen persufflation: orthotopic pig heart transplantations after 14-hr storage. AB - BACKGROUND: Improvement of heart preservation is still the greatest challenge in preservation research. The unchanged severe restriction of acceptable storage periods of heart grafts since the beginning of clinical heart transplantation indicates that technical innovations are necessary if a substantial improvement is to be achieved. METHODS: Here, we present the results of hypothermic preservation using the innovative technique of coronary oxygen persufflation (COP). COP simply adds gaseous oxygen to hypothermic graft storage and requires only a "valve guard" for reversible closure of the aortic valve. Fourteen-hr preservation was followed by orthotopic transplantation and evaluations of functional as well as metabolic recovery. Mature pig hearts, a model with restricted preservation tolerance similar to the human heart, were used to guarantee the clinical relevance of this study. RESULTS: After 14-hr hypothermic storage, COP-preserved hearts were able to recover within 2 hr of cardiopulmonary bypass to a steady cardiovascular function without mechanical or pharmacologic support. The left ventricular pressure amplitude of mHTK-COP-preserved hearts as well as energy charge potential recovered to pregrafting values and the ventricular power output to 66%. Hearts simply stored in University of Wisconsin (UW), modified Bretschneider's histidine-tryptophan-ketoglurate (mHTK), or Euro Flush with glutathione (EFG) solution had only limited recovery, with significantly lower ventricular power output of 18%, 29% or 30% of pregrafting controls on average. CONCLUSIONS: Fourteen-hr oxygenated pig heart preservation using COP results in optimal recovery. Storage preservation in solutions containing hyaluronidase (mHTK and EFG) results in higher recoveries as compared to UW solution, an effect that may support the excellent recovery after mHTK-COP preservation. PMID- 9869086 TI - Plasma exchange and tacrolimus-mycophenolate rescue for acute humoral rejection in kidney transplantation. AB - BACKGROUND: Acute renal allograft rejection associated with the development of donor-specific alloantibody (acute humoral rejection, AHR) typically carries a poor prognosis. The best treatment of this condition remains undefined. METHODS: During a 14-month period, 73 renal transplants were performed. During the first postoperative month, five recipients (6.8%) with AHR were identified. The diagnosis was based on: (1) evidence of severe rejection, resistant to steroid and antilymphocyte therapy; (2) typical pathologic features; and (3) demonstration of donor-specific alloantibody (DSA) in recipient's serum at the time of rejection. Pretransplant donor-specific T- and B-cell cross-matches were negative. RESULTS: Plasma exchange (PE, four to seven treatments per patient) significantly decreased circulating DSA to almost pretransplant levels in four of five patients, and improvement in renal function occurred in all patients. One patient had recurrent renal dysfunction in the setting of an increase in circulating DSA. A second series of five PE treatments decreased DSA and reversed the rejection episode. Rescue therapy with tacrolimus (initial mean dose: 0.14+/ 0.32 mg/kg/day) and mycophenolate mofetil (2 g/day) was used in five of five and four of five patients, respectively. With a mean follow-up of 19.6+/-5.6 months, patient and allograft survival are 100%. Renal function remains excellent with a mean current serum creatinine of 1.2+/-0.3 mg/dl. (range: 0.9-1.8 mg/dl). CONCLUSIONS: Our findings suggest that a therapeutic approach combining PE and tacrolimus-mycophenolate mofetil rescue has the potential to improve the outcome of AHR. PMID- 9869087 TI - Macrophage migration inhibitory factor expression in human renal allograft rejection. AB - BACKGROUND: Macrophage migration inhibitory factor (MIF) plays a pivotal role in immune-mediated diseases. Despite the long-standing association of MIF with the delayed-type hypersensitivity response, the potential role of MIF in allograft rejection is unknown. METHODS: MIF expression was assessed by in situ hybridization and immunohistochemistry staining in 62 biopsies of human renal allograft rejection and in normal human kidney. RESULTS: MIF mRNA and protein is constitutively expressed in normal kidney, being largely restricted to tubular epithelial cells, some glomerular epithelial cells, and vascular smooth muscle cells. In both acute and chronic renal allograft rejection, there was marked up regulation of MIF mRNA and protein expression by intrinsic kidney cells such as tubular epithelial cells and vascular endothelial and smooth muscle cells. There was also MIF expression by infiltrating macrophages and T cells. Of note, macrophage and T cell infiltrates were largely restricted to areas with marked up regulation of MIF expression, potentially contributing to the development of severe tubulitis and intimal or transmural arteritis. Quantitative analysis found that increased MIF expression in allograft rejection gave a highly significant correlation with macrophage and T cell accumulation in both the glomerulus and interstitium (P<0.001). In addition, the number of MIF+ tubules and interstitial MIF+ cells correlated significantly with the severity of allograft rejection (P<0.01), and the loss of renal function (P<0.01). In contrast, no up-regulation of renal MIF expression and no leukocyte accumulation was seen in allograft biopsies without evidence of rejection. CONCLUSIONS: This is the first study to demonstrate that local MIF expression is up-regulated during allograft rejection. The association between up-regulation of MIF expression, macrophage and T cell infiltration and the severity of renal allograft rejection suggests that MIF may be an important mediator in the process of allograft rejection. PMID- 9869088 TI - In situ enzymatic oligonucleotide amplification of hepatitis C virus-RNA in liver biopsy specimens (reverse transcriptase in situ polymerase chain reaction) after orthotopic liver transplantation for hepatitis C-related liver disease. AB - BACKGROUND: Hepatitis C infection recurs after orthotopic liver transplantation for hepatitis C virus (HCV)-related end-stage liver disease. Overlapping histopathologic features may present difficulties in differentiating recurrent HCV in the allograft from other conditions, especially rejection. METHODS: In this study, we evaluated the presence of HCV-RNA by reverse transcriptase in situ polymerase chain reaction (RT in situ RCR) in hepatic tissue, after orthotopic liver transplantation for HCV-related liver disease. Further, detection of HCV RNA was correlated with the serum HCV-RNA levels, histopathology, and clinical outcome. RESULTS: Twenty-five patients were part of this study. Seventeen patients were transplanted for HCV cirrhosis and eight for an underlying disease other than HCV. None of the patients in the non-HCV group had in situ RT-PCR detection of HCV-RNA. Positive RT in situ PCR for HCV was found in 9 of 17 HCV patients, and the patients had a clinical course consistent with recurrent hepatitis C disease. Four of these nine patients had an initial histologic diagnosis of rejection. The other eight patients in the HCV group had negative RT in situ PCR, and none of them had a course compatible with recurrent HCV disease, although four patients were histologically diagnosed as having chronic C hepatitis. The mean HCV-RNA level (log/mL) in the patients who had in situ detection of HCV-RNA was 7.01+/-0.26. Although RT-PCR was negative in 8 of 17 HCV patients, the patients were serologically viremic and the mean HCV-RNA level was 6.05+/-0.33 (P=0.03). CONCLUSIONS: Our findings indicate that the HCV in situ RT PCR assay may be helpful in the differentiation of recurrent hepatitis C disease from rejection. This may further help in the adjustment of immunosuppression. PMID- 9869090 TI - Intact pancreatic islet function despite humoral xenorecognition in the pig-to monkey combination. AB - BACKGROUND: The aim of this study was to analyze humoral xenoreactivity of various Old World primate species sera against pig islets and the effects of these sera on pig islet viability and function after culture. METHODS: Freshly isolated or cultured adult pig islets were analyzed by immunohistology or by cytofluorimetry for Old World primate xenoreactive natural antibody (XNA) binding and complement deposition. Complement-mediated cytotoxicity was evaluated by 51Cr release assays. After 4 days of culture in 50% sera from Old World primates, the morphology and in vitro metabolic function of pig islets were also analyzed. RESULTS: Chimpanzee, Macaca mulatta (rhesus), or baboon XNA binding was detectable only on intra-islet endothelial cells (ECs). Incubation of pig islets with sera from all Old World primate species tested showed C3 and C4 deposition on ECs and on some surrounding endocrine cells. However, membrane attack complex (MAC) showed a pattern of positivity similar to XNA binding, i.e., restricted to ECs only. No deposition of factor B was detected. Although complement cascade was activated, no cytotoxicity was observed after incubation of islets with chimpanzee serum, whereas between 10% and 35% 51Cr specific release was obtained with rhesus, baboon, or Macaca fascicularis sera. Despite this cytotoxic effect, purified pig islets showed a normal morphology and a well-preserved insulin release in response to an acute glucose stimulus, after prolonged culture with 50% serum obtained from all primate species considered. CONCLUSIONS: Despite the fact that pig beta-cell function was not affected by the serum of any of the primate species tested, some of them yielded significant lysis of islet cells, presumably as a result of a cytotoxic effect on intra-islet ECs. These data show that Old World primate sera from different species do not have equivalent effect on pig islets; these differences should be taken into account in preclinical trials of pig islet xenotransplantation. PMID- 9869089 TI - Intracellular expression in pig cells of anti-alpha1,3galactosyltransferase single-chain FV antibodies reduces Gal alpha1,3Gal expression and inhibits cytotoxicity mediated by anti-Gal xenoantibodies. AB - BACKGROUND: The carbohydrate structure Gal alpha1,3Gal expressed on pig cells is the major antigen recognized by xenoreactive natural antibodies in the higher primates. In xenotransplantation, natural antibodies binding to that structure initiate hyperacute rejection, and the anti-Gal alpha1,3Gal antibodies that are elicited probably take part in later phases of vascularized graft rejection. This epitope also appears to be involved in innate cellular responses. Inactivation of alpha1,3 galactosyltransferase in transgenic pigs would certainly lead to the success of xenotransplantation, but gene knockout in pigs is not feasible yet. METHODS: As a novel strategy to inhibit alpha1,3 galactosylation, we generated recombinant single-chain Fv (ScFv) antibodies directed against pig alpha1,3 galactosyltransferase and evaluated the effect of their intracellular expression on enzyme activity and Gal alpha1,3Gal expression. RESULTS: After in vitro transfection in pig cells, the scFv antibody anti-pig alpha1,3 galactosyltransferase reduced the amount or function of enzyme by up to 70% as evidenced by immunofluorescence and measurement of cell-associated activity. Consequently, Gal alpha1,3Gal on cell membranes was reduced to the same extent. This led to a profound (more than 90%) reduction in the cytotoxicity involving anti-Gal antibodies and complement. CONCLUSION: Although not sufficient to knock out the overall human anti-pig natural xenoreactivity, intracellular expression of the scFv antibody anti-alpha1,3-galactosyltransferase in pig cells significantly decreases the amount of Gal alpha1,3Gal and could be important to protect cells from elicited antibodies as well as from innate effectors. PMID- 9869091 TI - Heterogeneous expression of Gal alpha1-3Gal xenoantigen in pig kidney: a lectin and immunogold electron microscopic study. AB - BACKGROUND: The Gal alpha1-3Gal antigen (Gal alpha) is the primary target for human natural anti-pig xenoantibodies. The presence of Gal alpha has been shown in porcine endothelial cells (ECs) using light microscopy, whereas the expression of Gal alpha in other cell structures in the porcine kidney is only partially characterized. METHODS: Immunogold electron microscopy of pig kidney cryosections was performed using Griffonia simplicifolia isolectin B4 and affinity isolated human anti-Gal alpha1-3Gal antibodies. RESULTS: The most intense expression of Gal alpha was found on the apical and basolateral portions of the plasma membrane of the proximal convoluted tubule segments 1 and 2 cells, whereas segment 3 and 4 cells were negative. A strong staining was found in peritubular capillary ECs and in the inner medullary and papillary collecting duct cells. Moderate labeling of ECs and subendothelium was observed in large blood vessels, whereas glomerular ECs reacted weakly. Additionally, glomerular parietal epithelial cells, connecting tubule cells, and some cortical collecting duct cells were labeled. Among interstitial cells, a part of type-1 cells and all type-2 cells were labeled, whereas others were negative. CONCLUSIONS: By immune electron microscopy, a detailed information of the Gal alpha antigen distribution in porcine nephrons and blood vessels has been revealed, which clarifies conflicting data obtained by light microscopy. In addition, expression of the Gal alpha antigen in the renal interstitial cells was documented for the first time. These data are of importance for the understanding of xenoantibody-mediated hyperacute rejection, for interpretation of pig kidney xenograft biopsies, and for generating transgenic pigs lacking the Gal alpha epitope. PMID- 9869092 TI - Cytokine profiles of aqueous humor and graft in orthotopic mouse corneal transplantation. AB - BACKGROUND: Cytokine profile is a key in understanding the mechanisms of allograft rejection. Cytokine expression in the aqueous humor and the correlation between the aqueous humor cells and corneal infiltrating cells are not fully understood in corneal transplantation. METHODS: Orthotopic mouse corneal transplantation was performed using BALB/c (H2d) mice as recipients, and C3H/He (H2k) and BALB/c mice as donors for allografts and isografts, respectively. Immunocytochemistry was performed on aqueous humor cells. Corneal graft was studied immunohistochemically. Cytokine gene expressions of the cells infiltrating the aqueous humor and corneal grafts were determined by the semiquantitative reverse transcription and polymerase chain reaction method. RESULTS: Interferon-gamma, interleukin (IL)-2, IL-4, and IL-10 were detected in the cells infiltrating the aqueous humor and corneal grafts at both the protein and gene expression levels. T helper 1 (Th1) cytokine expressions at the protein level, however, were consistently predominant in the rejected allografts compared to those of Th2 cytokines. The cytokine and surface marker profiles of the cells in the aqueous humor corresponded well to those of the cells infiltrating the corneal grafts. Cytokine protein and mRNA expression levels in the aqueous humor decreased rapidly. CONCLUSIONS: Allorejection in corneal transplantation is Th1 cytokine-predominant. Infiltrating cells do not express Th2 cytokine so much in allograft rejection, as compared with Th1 cytokine. The cell infiltration patterns of the aqueous humor were well correlated with those of the cornea. PMID- 9869093 TI - Redefining peripheral tolerance in the BALB/c to CBA mouse cardiac allograft model: vascular and cytokine analysis after transient CD4 T cell depletion. AB - BACKGROUND: To evaluate cardiac allografts from recipients that had achieved peripheral tolerance after transient CD4+ T cell depletion, we analyzed cellular infiltrate, cytokine expression, and vascular thickening. Long-surviving cardiac allografts from tolerant recipients were compared with acutely rejecting allografts and isografts. METHODS AND RESULTS: In CBA mice treated with anti-CD4 (GK1.5, 0.5 mg intraperitoneally on days 1-28), BALB/c cardiac allografts survived >100 days. These recipients were tested for tolerance at >70 days, by challenge with donor and third-party (C57BL/6) skin grafts. BALB/c skin grafts survived >30 days, although C57BL/6 skin was rejected in <12 days, reflecting alloantigen-specific peripheral tolerance. When vascular thickening in graft arteries was assessed and computerized measurements performed, heart allografts from tolerant recipients showed significantly increased percentage of luminal occlusion compared with isografts (47% compared with 1.2%). Semiquantitative reverse transcriptase-polymerase chain reaction was used to assess normalized intragraft mRNA transcripts for cytokines and T cell markers, with immunoperoxidase staining of frozen sections to confirmed the presence of protein. Compared with rejecting grafts, well-preserved hearts from tolerant mice had lower levels of macrophage and T cell infiltration and decreased transcription of interferon-gamma, interleukin (IL)-2, IL-10, and inducible nitric oxide synthase. IL-4 expression was similar in both groups. CONCLUSIONS: The degree of tolerance achieved allowed specific acceptance of donor skin grafts, preserved primary graft function, and reduced inflammatory activation. Tolerance did not, however, completely prevent macrophage and T cell infiltration of the graft or the development of vascular lesions typical of chronic rejection. PMID- 9869094 TI - Effect of a topically applied neutralizing antibody against vascular endothelial growth factor on corneal allograft rejection of rat. AB - BACKGROUND: Studies in corneal transplant rejection remain important because acute immunologic rejection continues to be the leading cause of human corneal transplant failure. As the permeability of vessels and the neovascularization induce cells infiltration into the graft, we considered the possibility that vascular endothelial growth factor (VEGF), a potent permeability-increasing factor and angiogenesis-mediating factor, could participate in the immune response. METHODS: As the established corneal transplant model for rejection, the corneal transplant between Lewis and Fisher rats has been reported. First, we evaluated VEGF production in the graft by immunohistochemical method in the animal model. Next, we tried to neutralize the effect of VEGF by topical administration of anti-VEGF antibody. We administered anti-VEGF antibody as eye drops for 10 days just after the transplantation of the established animal corneal transplant model. RESULTS: VEGF was strongly produced from the infiltrative cells into the graft. Anti-VEGF antibody significantly suppressed the acute rejection compared with saline or rabbit IgG. CONCLUSIONS: The inhibition of VEGF by topically applied neutralizing antibody is a new potential therapeutic strategy for the treatment of corneal transplantation. PMID- 9869095 TI - Specific expression of Epstein-Barr virus in cutaneous squamous cell carcinomas from heart transplant recipients. AB - BACKGROUND: We investigated a Swedish group of 114 immunosuppressed cardiac allograft patients for the occurrence of posttransplant cutaneous squamous cell carcinomas. A total of 15 tumors were detected in specimens from 5 patients. METHODS: The tumors were analyzed for the presence of Epstein-Barr virus (EBV) genomes as well as EBV-specific gene expression by using three different techniques; the polymerase chain reaction (PCR), in situ hybridization, and immunohistochemistry. The material was also tested by PCR for high-risk human papilloma virus genome. RESULTS: EBV DNA could be detected by PCR in 10 of the investigated tumors, 7 of which also expressed EBV latent membrane protein 1 and/or EBV-encoded RNAs. No EBV genomes or EBV gene products could be detected in normal skin/resection margins, available from three of the tumors investigated. All tumors were negative for high-risk human papilloma virus DNA analyzed by PCR. CONCLUSIONS: In this study, we have found a high incidence of EBV-specific expression in posttransplant cutaneous squamous cell carcinomas. These results suggest that at least some of the skin cancers developing in immunocompromised heart transplant recipients are associated with EBV. PMID- 9869096 TI - Apoptosis of T lymphocytes in liver and/or small bowel allografts during tolerance induction. AB - BACKGROUND: Apoptosis of parenchymal cells has been described during allograft rejection. Immunologically privileged tissue in the mouse has been found to prevent rejection by initiating apoptosis of infiltrating lymphocytes. The aim of this study was to investigate whether apoptosis may play a role in T-cell regulation during rejection and subsequent tolerance induction after liver transplantation (LTx) and combined liver/small bowel transplantation (LSBTx). METHODS: LTx and LSBTx (Brown Norway-->Lewis) were performed without immunosuppression. Cell migration, activation, and apoptosis were investigated by means of sequential histology, immunohistochemistry, and the terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling assay. Donor (Brown Norway) and third-party (Dark Agouti) cardiac allografts were transplanted into LSBTx recipients to determine specific tolerance. RESULTS: Transient acute cellular rejection occurred after LTx and LSBTx and was followed by specific tolerance. The kinetics of apoptosis were similar in liver allografts after LTx and LSBTx, but differed from the processes in small bowel allografts after LSBTx. Apoptosis of parenchymal cells in the grafted livers correlated directly with interleukin-2 receptor expression of the infiltrating T cells. During the late phase of rejection, a peak of apoptosis in the lymphocyte infiltrate was demonstrated, characterized as predominantly apoptotic CD8+ T lymphocytes. CONCLUSIONS: These results demonstrate that specific tolerance is achieved in both LTx and LSBTx after a transient rejection crisis. Apoptosis is involved in graft rejection and tolerance induction. Activation of T lymphocytes correlates with parenchymal cell apoptosis in the allograft. T-cell inactivation seems to result in apoptosis of cytotoxic T cells and tolerance, which appears to be unique to the liver allograft. PMID- 9869097 TI - Chronic rejection of major histocompatibility complex class II-disparate skin grafts after anti-CD3 therapy: a model of antibody-independent transplant vasculopathy. AB - BACKGROUND: Chronic rejection remains a leading cause of allograft loss. Histologically, it is characterized by arterial intimal thickening and parenchymal fibrosis. The immune mechanisms triggering chronic rejection are still uncompletely understood. METHODS: We performed major histocompatibility complex (MHC) class H-incompatible skin grafts from C-H2bm12 (bm12, H2bm12) into C57BL/6 (C57BL/6, H2b) recipients immunosuppressed with a short course of anti CD3 monoclonal antibodies to prevent acute rejection. RESULTS: More than 80% of grafts survived for prolonged periods, but eventually all displayed macroscopic and microscopic evidence of chronic rejection. At histology, there was a progressive arterial intimal thickening as well as intense dermal fibrosis. This was accompanied by an inflammatory infiltrate consisting of lymphocytes and macrophages, but also of a considerable number of eosinophils. Mice with chronic rejection were unable to generate anti-donor MHC class II cytotoxic T lymphocyte activity at either 20 or 60 days after transplant. Furthermore, transplantation of bm12 skins on C57BL/6-congenic, Ig knock-out mice was associated with the development of a chronic rejection that was identical to that occurring in wild type C57BL/6 animals, indicating that alloantibodies are not necessary in this model. CONCLUSIONS: (1) Skin grafts may undergo chronic rejection with the characteristic lesions of vasculopathy and fibrosis; (2) chronic rejection of MHC class II-disparate skins may occur in the absence of direct cytotoxic T lymphocyte activity or alloantibodies. PMID- 9869098 TI - Adenovirus-mediated gene transfer to liver grafts: an improved method to maximize infectivity. AB - BACKGROUND: Adenoviral gene therapy in liver transplantation has many potential applications, but current vector delivery methods to grafts lack efficiency and require high titers. In this study, we attempted to improve gene delivery efficacy using three different delivery methods to liver grafts with adenoviral vector encoding the LacZ marker gene (AdLacZ). METHODS: AdLacZ was delivered to cold preserved rat liver grafts by: (1) continuous perfusion via the portal vein (portal perfusion), (2) continuous perfusion via both the portal vein and hepatic artery (dual perfusion), and (3) trapping viral perfusate in the liver vasculature by clamping outflow (clamp technique). RESULTS: Using 1x10(9) plaque forming units of Ad-LacZ (multiplicity of infection of 0.4), transduction rate in 3-hr preserved liver grafts, determined by 5-bromo-4-chromo-3-indolyl-beta-D galactopyranoside staining and beta-galactosidase assay 48 hr after transplantation, was best with clamp technique (21.5+/-2.7% 5-bromo-4-chromo-3 indolyl-beta-D-galactopyranoside-positive cells and 81.1+/-3.6 U/g beta galactosidase), followed by dual perfusion (18.5+/-1.8%, 66.6+/-19.4 U/g) and portal perfusion (8.8+/-2.5%, 19.7+/-15.4 U/g). Further studies using clamp technique demonstrated a near-maximal gene transfer rate of 30% at multiplicity of infection of 0.4 with prolonged cold ischemia to 18 hr. Transgene expression was stable for 2 weeks and slowly declined to 7.8+/-12.1% at day 28. Lack of inflammatory response was confirmed by histopathological examination and liver enzymes. Transduction was selectively induced in hepatocytes with nearly no extrahepatic transgene expression in the lung and spleen. CONCLUSIONS: The clamp technique provides a highly efficient viral gene delivery method to cold preserved liver grafts. This method offers maximal infectivity of adenoviral vector with minimal technical manipulation. PMID- 9869099 TI - Chemokine receptor (CXCR4) mRNA-expressing leukocytes are increased in human renal allograft rejection. AB - BACKGROUND: Mononuclear cell infiltration is a common feature of cell-mediated renal transplant rejection. Chemokines and their corresponding receptors likely play a central role in directing specific classes of leukocytes to graft sites during rejection. Localization of chemokine receptors may help us understand how specificity in leukocyte trafficking is achieved in renal inflammatory processes. The localization of the chemokine receptor CXCR4 in human kidney and in renal transplant rejection is unknown. METHODS: We generated a riboprobe specific for the detection of CXCR4 mRNA by in situ hybridization to evaluate cellular sites of synthesis of this receptor in native human kidneys (n=11) and in human allograft nephrectomies with features of severe rejection (n=14). RESULTS: By in situ hybridization, CXCR4 mRNA expression is undetectable in intrinsic glomerular, tubular, and renovascular cells in native kidneys. When renal interstitial inflammation is present, CXCR4 mRNA expression is localized to a large fraction of infiltrating leukocytes. Large numbers of CXCR4-expressing cells are detected in cell-mediated renal allograft rejection. Double immunolabeling for CD3 antigen identified a large fraction of infiltrating CXCR4 mRNA-expressing cells as T lymphocytes. CXCR4 mRNA-expressing cells were frequently seen in neointimal lesions of vascular rejection in allograft nephrectomies. CXCR4 mRNA expression was identified in infiltrating neointimal T lymphocytes, but not smooth muscle cells by immunolabeling. CONCLUSIONS: We demonstrate the involvement of CXCR4 mRNA-expressing infiltrating cells in human renal interstitial and vascular allograft rejection. Signaling via the CXCR4 receptor may be one mechanism by which chemokines mediate leukocyte trafficking in renal allograft rejection. PMID- 9869100 TI - Prolongation of porcine islet xenograft survival in mice after therapy with immunosuppressive peptides. AB - BACKGROUND: Recently, peptides derived from the heavy chain of HLA-B2702 have been shown to modulate immune responses. In this study, we examined the use of these peptides for immunosuppression in a pig to mouse islet xenograft model. METHODS: Purified porcine islets were transplanted in autoimmune (non-obese diabetic) and non-autoimmune (streptozotocin-injected CBA or C57/Bl6) diabetic mice. Various dosing regimens of HLA-derived peptides with and without antilymphocyte therapy were administered to recipient mice. Graft rejection was determined by daily serum glucose determinations, and, at selected time points, grafts were removed to demonstrate function and provide immunohistochemical examination. RESULTS: HLA-derived peptides were demonstrated to prolong graft survival in both pretransplant and posttransplant treatment regimens. This effect was increased with concomitant antilymphocyte therapy. CONCLUSIONS: Further elucidation of the mechanism of action of these immunomodulatory peptides may help in the development of novel immunosuppressive protocols. PMID- 9869101 TI - Cytomegalovirus colitis mimicking a colonic neoplasm or ischemic colitis 4 years after heart transplantation. AB - BACKGROUND: Cytomegalovirus (CMV) colitis is a polymorphous disease presenting in immunodepressed patients in a variety of clinical forms that can delay diagnosis and therapy. We report the case of a patient who presented with abdominal pain 4 years after heart transplantation; clinical and x-ray findings were suggestive of a neoplastic or ischemic stenosis, and histopathological examination likewise initially suggested an ischemic etiology. METHODS: Tissue samples were fixed in 10% formaldehyde, embedded in paraffin, cut, and stained with hematoxylin/eosin and periodic acid-Schiff-Alcian Blue. Immunohistochemistry with monoclonal antibodies was performed using an indirect immunoperoxidase method. RESULTS: CMV colitis was eventually diagnosed and resolved with surgery and specific anti-CMV therapy. CONCLUSIONS: CMV colitis should be suspected in any heart transplant patient with signs or symptoms of abdominal pathology, even without classical signs or symptoms of CMV infection. If stenotic lesions are present, surgery may be required not only to remove the obstruction but also to rule out malignancy. PMID- 9869102 TI - Higher spontaneous apoptotic index in small cell compared with non-small cell lung carcinoma cell lines; lack of correlation with Bcl-2/Bax. AB - Spontaneous apoptosis was assessed in ten small-cell (SCLC) and five non-small cell (NSCLC) lung carcinoma cell lines by the TUNEL assay and chromatin cleavage. TUNEL staining showed significantly higher apoptotic index (AI) in SCLC (2-20%) compared with NSCLC lines (0.2-1%) in untreated exponentially growing cells. Six out of ten SCLC and none of the NSCLC showed DNA fragmentation when analysed by agarose gel electrophoresis. Field inversion pulse gel electrophoresis was used in a subset of cell lines and showed the presence of high molecular weight fragments in untreated SCLC lines U-1285 and U-1906 cells, but not in the NSCLC line U-1810. Important molecular determinants of apoptosis were studied by Western blot. Bcl-2 was detected at highest level in SCLC. There was no correlation between the ratio Bcl-2/Bax and AI in all tested cell lines. Neither p53 nor c-Myc protein status correlated to AI. Pro-caspase-3 was expressed in all cell lines without correlation to AI and no difference between the SCLC and NSCLC groups was found. In conclusion, this study shows a high degree of spontaneous apoptosis in SCLC lines compared to NSCLC lines unrelated to Bcl-2/Bax ratio. PMID- 9869103 TI - Effect of cigarette smoking on major histological types of lung cancer in men. AB - This study examined the effect of cigarette smoking parameters such as intensity, duration, age at initiation, and quitting on the development of different histological types of lung cancer in men. We used data from a case-control study conducted in Philadelphia between 1985 and 1987. Cases included 482 men with histologically confirmed lung cancer diagnosed in 15 selected hospitals in Philadelphia. Controls were selected from a neighborhood survey of men in Philadelphia conducted concurrently to the case recruitment. Most aspects of smoking were associated with all the major histological types of lung cancer. Number of cigarettes smoked per day was the strongest predictor of risk of developing lung cancer. Early age at initiation of smoking significantly increased the risk of small cell carcinoma (odds ratio = 3.0; 95% CI, 1.1-8.4). Quitting smoking reduced the risk of squamous cell and adenocarcinoma; however, it did not affect the risk of small cell lung cancer. The findings of this study suggest the need for greater emphasis on smoking prevention programs, especially in adolescents. PMID- 9869104 TI - Extent of mediastinal node metastasis in clinical stage I non-small-cell lung cancer: the role of systematic nodal dissection. AB - OBJECTIVE: To determine the extent of lymph node metastasis in clinical stage I non-small-cell lung cancer (NSCLC). METHODS: We performed a retrospective review of 524 patients with clinical stage I NSCLC who underwent lobectomy with systematic nodal dissection. RESULTS: The nodal status was N0 in 409 patients (78%), N1 in 44 (8%), N2 in 67 (13%), and N3 in four (0.8%). Thirty-six patients had single-level mediastinal nodal metastases and 35 had multi-level metastases. The incidence of N2/3 disease in patients with adenocarcinoma/squamous cell carcinoma/other histologic types according to tumor size was 0/0/0%, respectively, in tumors < or = 10 mm in diameter, 12/0/0% in tumors 11-20 mm in diameter, 14/4/23% in tumors 21-30 mm in diameter, and 26/14/20% in tumors >30 mm in diameter. Nodal metastases to the upper mediastinum from middle or lower lobe lesions were frequently observed in 51 N2 adenocarcinomas, whereas those to the lower mediastinum from upper lobe lesions were rare. Of 10 N2 squamous cell carcinomas, seven had regional and three had non-regional nodal metastases. The 5 year survival rate was 68, 43, and 30% in N0, N1, and N2, respectively (P<0.01, N0 versus N1, N0 and N2). CONCLUSIONS: Systematic mediastinal nodal dissection should be routinely performed for clinical stage I lung cancer to ensure the correct nodal status, but it might be dispensable in the patients with peripheral squamous cell carcinoma < or = 20 mm in diameter, with central squamous cell carcinoma < or = 30 mm, and with adenocarcinoma < or = 10 mm. When systematic nodal dissection cannot be performed, the incidence and extent of nodal metastases should be taken account with respect to histologic type, size, and location of the tumor. PMID- 9869105 TI - Multicenter randomized clinical trial on high-dose epirubicin plus cis-platinum versus vinorelbine plus cis-platinum in advanced non small cell lung cancer. AB - BACKGROUND: High dose Epirubicin (HD-EPI) (>90 mg/m2) and Vinorelbine (VNR) demonstrated antitumor activity as single agent (about 20%) in the treatment of advanced NSCLC. This trial compares these two agents combined with cisplatin (CP). PATIENTS AND METHODS: From August 1992 to February 1996, 228 patients with locally advanced or metastatic NSCLC were randomized to receive either EPI 120 mg/m2 as i.v. bolus plus Cisplatin (CP) 60 mg/m2 on day 1 (regimen A) or VNR 25 mg/m2 as i.v. bolus on day 1 and 8 plus CP 60 mg/m2 on day 1 (regimen B). Both treatments were recycled every 21 days up to a maximum cumulative dose of EPI of 840 mg/m2 or 12 cycles. Eligible patients were 212 and 198 patients were evaluable for objective response (95 in arm A and 103 in arm B). The main characteristics of eligible patients were: male/female 179/33; median age 61 (42 72); median Karnofsky PS 80 (70-100); stage IIIA 12%, stage IIIB 40%, stage IV 41%, recurrence 7%; histotype: epidermoid 48%, adenoca 36%, others 16%. RESULTS: The following response rates were observed in regimens A and B, respectively; CR, 1 and 2%, PR, 32 and 25% (P = 0.4567). Median CR + PR duration was 9 and 8 months, respectively. Median survival was 10.5 and 9.6 months, respectively. Grade III-IV leucopenia occurred in 38 and 21% in arm A and arm B, respectively(P = 0.01), thrombocytopenia in 6 and 0% (P = 0.02), anemia in 8 and 7% (n.s.). Non hematological toxicity was moderate and the only difference between the treatments was alopecia (88 vs. 33% in arm A and B, respectively). Supraventricular arrhythmia occurred in three patients on regimen A; a >15% LVEF absolute decrease was observed in 9 (22.5%) and three (14%) patients on arm A and arm B, respectively (n.s.). No congestive heart failure was observed. CONCLUSION: HD-EPI+CP and VNR+CP are both active combinations in advanced NSCLC with a similar response rate, response duration and survival but regimen A was significantly more toxic (myelosuppression and alopecia). PMID- 9869107 TI - Sequential treatment with vindesine-ifosfamide-platinum (VIP) chemotherapy followed by platinum sensitized radiotherapy in stage IIIB non-small cell lung cancer: a phase II trial. AB - PURPOSE: Daily administration of cisplatin concomitant with radiotherapy improved the overall survival in inoperable non-small cell lung cancer (NSCLC) in one EORTC study. In this study, we prospectively investigated the efficacy and toxicity of a sequential treatment with three cycles of vindesine-ifosfamide platinum (VIP) induction chemotherapy, followed by daily cisplatin-sensitized radiotherapy. METHODS: Between June 1993 and June 1995, 23 previously untreated patients with stage IIIB NSCLC with World Health Organization performance status 0 or 1 were included. Chemotherapy consisted of platinum 30 mg/m2 and ifosfamide 1200 mg/m2 i.v. on days 1, 2 and 3, and vindesine 3 mg/m2 i.v. on days 1 and 8, every 4 weeks. After three cycles and at least stable disease, radiotherapy was started (30 Gy in 10 fractions, followed by a boost of 22 Gy in 10 fractions). Each fraction was preceded by Platinum 6 mg/m2 i.v. RESULTS: Nineteen patients completed the sequential therapy. One patient died from neutropenic sepsis during the first cycle of chemotherapy, and three patients had progressive disease after chemotherapy. The overall response rate after sequential therapy was 47% (95% confidence interval 24-80), median survival was 10.6 months, 1- and 2-year survival rates were 47 and 16%, respectively. Major toxicity consisted of neurotoxicity grade III-IV in 18% and of leukopenia grade III-IV in 22% of the patients. Acute radiation pneumonitis grade III occurred in 11% of the patients. CONCLUSION: Three-drug VIP induction chemotherapy followed by cisplatin sensitized radiotherapy is feasible, with acceptable, albeit substantial, toxicity. In spite of the theoretically promising sequence of therapies, survival results remain disappointingly low. PMID- 9869106 TI - Phase II study of hyperfractionated radiotherapy and concurrent weekly alternating chemotherapy in limited-stage small cell lung cancer. AB - Despite recent advances in combined modality therapy, long-term survival remains elusive in most patients with limited-stage small cell lung cancer (SCLC). The present study was designed to evaluate the activity and toxicity of concurrent hyperfractionated radiotherapy and weekly, alternating-regimen chemotherapy. Twelve patients with limited-stage SCLC and performance status 0-1 were treated with cyclophosphamide 250 mg/m2, etoposide 100 mg/m2, and cisplatin 50 mg/m2 on day 1 every other week, and vincristine 1 mg/m2 on day 8, and ifosfamide 1.2 mg/m2 on days 8 and 9 every other week. Hyperfractionated thoracic radiotherapy, consisting of three daily doses of 1.1 Gy for 20 days to a total dose of 66 Gy, was started on day 1 of chemotherapy. Ten patients (83%) exhibited an objective response (9 CRs and 1 PR) with a median duration of response of 8.6 months. Two complete responders died at 50 and 53 months without evidence of progression and two remain alive and free of SCLC at 73 and 87 months. Median survival was 19.8 months with 2- and 5-year survival rates of 50 and 17%, respectively. Severe toxicity, including grade 3-4 esophagitis (67%) and granulocytopenia (83%), as well as debilitating fatigue and pneumonitis, prompted early termination of the trial. Hyperfractionated radiotherapy and concurrent weekly alternating-regimen chemotherapy resulted in promising response and survival rates, but induced excessive toxicity, in patients with limited-stage SCLC. PMID- 9869109 TI - Lung health and the International Union Against Tuberculosis and Lung Disease. PMID- 9869108 TI - Malignant neuroendocrine tumor presenting a huge mediastinal mass controlled with radiation therapy. AB - A case of malignant neuroendocrine tumor presenting a huge mediastinal mass controlled with radiation therapy is reported. A thoracotomy was performed on a 57-year-old male and a mass was found to invade the trachea, main bronchus, and left atrium. The tumor was unresectable. Subsequently, cisplatin was administered, but the tumor grew in size and radiation therapy was recommended. The tumor responded well to thoracic irradiation (56 Gy) and disappeared. Three months later, lymph nodes metastases were noted in the retrocrural and upper abdominal paraaortic regions, but these were controlled again with palliative irradiation. Without further treatment, he survived free of disease for over 5 years. Specimens obtained during the thoracotomy showed that the tumor consisted mainly of small round cells with a rosette formation. Immunohistochemically the tumor was positive to NSE and slightly positive to keratin, but negative to LCA, L26, UCLH-1, EMA, Leu7, and chromogranin, suggesting a malignant tumor derived from neuroendocrine tissue. PMID- 9869110 TI - The International Journal of Tuberculosis and Lung Disease--a home for respiratory epidemiology. PMID- 9869111 TI - DNA vaccines: application to tuberculosis. AB - Since the first observations in the early 1990s, the scientific literature on DNA vaccines has been growing exponentially. This article reviews the history and general principle, summarizes current knowledge on immune mechanisms, discusses safety considerations and highlights possible advantages of this technique as compared to the classic vaccines. Special emphasis is placed on the potential of DNA vaccines with respect to tuberculosis. PMID- 9869112 TI - Acceptance and safety of directly observed versus self-administered isoniazid preventive therapy in aboriginal peoples in British Columbia. AB - OBJECTIVE: To document experience with directly observed chemoprophylaxis (DOPT) compared to self-administered isoniazid (INH) among aboriginal persons in British Columbia. DESIGN: DOPT was compared to self-administered delivery (SAD) over a 3 year period. All aboriginal persons who received INH chemoprophylaxis in British Columbia between 1992 and 1994 were evaluated. Therapy completion rates and adverse outcomes associated with SAD were compared with DOPT. Treatment allocation was by patient choice. RESULTS: Of 608 people who received INH prophylaxis, 443 received SAD (mean age 31.6 years) and 165 received DOPT (mean age 23.9 years). Two hundred and seventy (60.9%) SAD compared to 124 (75.2%) in the DOPT group completed 6 months of INH (P = 0.0011). The 12-month completion rates were 162/443 (36.6%) for the SAD group and 84/165 (50.9%) for the DOPT group (P = 0.0014). Adverse reactions requiring discontinuation of medication occurred in 13.5% of the patients on SAD and 9.7% of those receiving DOPT (P = 0.202). The most common reason cited for failure to complete therapy was non cooperation in both groups. There were three deaths in the SAD group, one of which was due to suicide by self-ingestion of INH. CONCLUSIONS: These data demonstrate that in aboriginal people compliance with preventive therapy can be improved by DOPT. Non random allocation to treatment groups might have influenced our findings, and further prospective randomized trials and cost-effectiveness analyses are required. PMID- 9869113 TI - Preventive therapy for tuberculosis in Western Australia. AB - SETTING: State Tuberculosis Control Programme, Western Australia. OBJECTIVE: To ascertain baseline information, applicability and efficacy of preventive therapy for tuberculosis (TB) under indirectly supervised treatment in Western Australia. DESIGN: Retrospective analyses of records of persons with TB infection who were prescribed preventive therapy for the period 1993-1996 inclusive, using simple descriptive statistics. RESULTS: Preventive therapy was given to 411 persons after screening for TB due to on-arrival migrant surveillance (269), contact tracing (59), tuberculin surveys (59) and other reasons (24). Six-month isoniazid monotherapy (HMT) was prescribed for 403 and multidrug regimens for the remaining eight. Excepting 34 whose compliance was unknown, varying degrees of treatment non-adherence were found in 90 (24%) of the remaining 369 persons given HMT, including 36 (10%) with under five months of total medication. Minor adverse drug effects occurred in 32 (9%) subjects and contributed to the non-adherence in 23 of these. One person has since developed active TB. CONCLUSIONS: Preventive therapy in Western Australia conformed to generally accepted guidelines. Varying degrees of non-adherence to HMT occurred in 24% of persons, but 90% completed adequate therapy under indirect supervision. Non-adherence is significantly related to adverse drug effects. PMID- 9869114 TI - Risk of tuberculosis in dialysis patients: a population-based study. AB - SETTING: Provincial tuberculosis (TB) and dialysis registries. OBJECTIVE: To document the risk of TB among patients on dialysis and to describe the clinical characteristics of these cases. METHODS: All cases of TB occurring among dialysis patients in British Columbia between January 1990 and December 1994 were reviewed, as were the age-specific rates for TB among the general population during the same period. RESULTS: During that period, a total of seven cases of TB occurred among 560 patients on hemo-dialysis and two cases among 326 patients on peritoneal dialysis. On an annual basis the rate of TB in the dialysis population was 253 per 100,000, compared to an age-matched rate of 10.1 per 100,000 in the general population, giving a relative risk of 25.3 (95% confidence interval 22.86 31.49, P = 0.0000001). CONCLUSIONS: The risk of TB in dialysis patients is significantly higher than previously reported from non population-based studies, indicating that dialysis patients should be considered a possible target population for systematic evaluation for the presence of TB infection and consideration for chemoprophylaxis. PMID- 9869115 TI - Tuberculosis control in Bangladesh: success of the DOTS strategy. AB - SETTING: Tuberculosis (TB) has been a major public-health problem in Bangladesh for many decades. National control efforts in the past have not been successful, with less than half of detected cases being cured. In 1993, a project based on the DOTS (directly observed treatment, short-course) strategy was initiated for a population of approximately one million in a rural setting. Following a 78% cure rate in the initial cohort of new smear-positive patients, the project was expanded in phases to cover a rural population of 67 million in 1996. OBJECTIVES: Routine programme data on all new sputum smear-positive patients registered in the TB project since its inception until 1996 were analysed. Case finding results are presented until 1996, as are results of sputum smear conversion after 2 months of treatment in new smear-positive patients for the same cohort of patients. Final treatment outcome results were analysed for new smear-positive patients registered up to 1995. RESULTS: A total of 41,525 patients were registered in the project during the 3-year period. Two-thirds of these were new smear-positive cases and 27% were new smear-negative patients. Sputum smear conversion in 26,151 new smear-positive patients at 2 months was 85%; 5% remained smear-positive, 3% had died and the rest had no sputum examination. Final treatment outcome results in 10,142 new smear-positive patients registered during 1993-1995 showed that 75% were cured, 4% completed treatment but did not have a sputum smear result, 2% remained smear-positive, 6% died, 10% defaulted and 3% were transferred out. CONCLUSION: The DOTS strategy can be successfully implemented in phases in large countries with a high tuberculosis burden. This success is due to decentralizing sputum smear microscopy and treatment delivery services to peripheral health facilities, utilizing the existing primary health care network. High cure rates can be maintained despite rapid expansion of coverage, with proper implementation of the strategy and regular monitoring of reports on case finding, sputum smear conversion and treatment outcome. Case detection needs to be further increased by informing and involving the community in TB control efforts through social mobilization. PMID- 9869116 TI - Management of pulmonary tuberculosis suspects with negative sputum smears and normal or minimally abnormal chest radiographs in resource-poor settings. AB - SETTING: Queen Elizabeth Central Hospital, Blantyre, Malawi. OBJECTIVES: 1) To determine the proportion of pulmonary tuberculosis (PTB) suspects with negative sputum smears and a normal/minimally abnormal chest radiograph (CXR) who are culture-positive for Mycobacterium tuberculosis, and 2) to determine how many develop smear or radiographic evidence of PTB (TB CXR) during follow-up. METHODS: PTB suspects with negative sputum smears and a normal/minimally abnormal CXR were given a second course of antibiotics and followed up at 3-week intervals over 3 months with repeat sputum smears and chest radiography. RESULTS: Of 79 patients (38 men and 41 women, mean age 33 years) with negative smears and a normal/minimally abnormal CXR, 16 (21%) were culture-positive for M. tuberculosis. Of 15 culture-positive patients who were alive and attended follow up, seven (47%) developed a TB-CXR by 3 months. Of 41 culture-negative patients who were alive and attended follow-up, 13 (32%) developed a TB-CXR, including one patient who became sputum smear-positive. TB-CXRs were found only in patients with a cough. CONCLUSION: TB suspects with negative smears and normal/minimally abnormal CXRs in high human immunodeficiency virus (HIV) prevalent countries should be given a second course of antibiotics. If cough improves, patients can be advised not to return for further follow-up. If cough continues, patients should return for further follow-up with sputum smear examination and chest radiography. Approximately 50% of those who have culture-positive PTB will develop a TB-CXR by 3 months and can be identified if radiographic facilities are available. PMID- 9869117 TI - Awareness about tuberculosis among nurses working in a tuberculosis hospital and in a general hospital in Delhi, India. AB - SETTING: A tuberculosis institute and a general hospital in Delhi, India. OBJECTIVE: To investigate the awareness of nurses about tuberculosis and to evaluate the differences in awareness, if any, between nurses working in tuberculosis and those in a general hospital. DESIGN: A pretested questionnaire survey was performed on 213 nurses. RESULTS: The present study showed that a substantial number of nurses have inadequate knowledge regarding causative factors, the importance of sputum examination, correct doses of routinely used short-course chemotherapy drugs, the minimum duration of short-course chemotherapy, instructions at discharge, and health education for patients and family members. If responding correctly to 75% of the questions asked is taken as the criterion for satisfactory awareness, only 40.2% of tuberculosis nurses and 10.7% of general hospital nurses had a satisfactory level of awareness. There was no effect of increasing age or years of experience on the level of awareness. CONCLUSIONS: There is a general lack of knowledge regarding various aspects of tuberculosis among nurses. Active interventions are required to improve awareness for a better implementation of the revised national tuberculosis control programme in India. PMID- 9869118 TI - Evaluation of a colorimetric assay based on 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) for rapid detection of rifampicin resistance in Mycobacterium tuberculosis. AB - OBJECTIVES: To standardise the colorimetric assay based on 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) for the rapid detection of rifampicin-resistant Mycobacterium tuberculosis in clinical practice and to evaluate the assay on a collection of 92 clinical isolates. DESIGN: The Bactec method was used as the reference method. Rifampicin was used for the susceptibility testing in the Bactec method at a concentration of 2 microg/ml. The MTT assay was performed in tubes containing 3 ml Dubos broth; the assay is based on the principle that live cells convert the yellow tetrazolium salt into a blue formazan. A final concentration of 2 microg/ml rifampicin was used in the assay. Optical density (OD) values at 570 nm were recorded on the third and sixth day. A strain was defined as susceptible when the relative optical density unit (RODU) (i.e., OD of rifampicin containing tube/OD of undiluted control) was < or = 0.2, and when the OD value of the rifampicin-containing tube on the sixth day was lower than the OD value on the third day. A strain was defined as resistant when the RODU was more than 0.5, and when there was an increase in OD value in the rifampicin-containing tube on the sixth day. The tubes were also read visually. RESULTS AND CONCLUSION: The result obtained by the MTT assay perfectly matched the result obtained by the Bactec method. The MTT assay was also interpretable by the naked eye. This simple, inexpensive assay could be used as a rapid screening method for identification of rifampicin-resistant strains in low income countries. PMID- 9869119 TI - Environmentally-induced malignant pleural mesothelioma and HLA distribution in Turkey. AB - SETTING: A large university hospital in Ankara, Turkey. OBJECTIVE: To investigate the potential links, if any, between the occurrence of malignant pleural mesothelioma (MPM) and the presence and distribution of human leukocyte antigens (HLA) in patients environmentally exposed to asbestos and erionite in rural Anatolia, Turkey. DESIGN: A case-control study design was used to compare the relative frequency and distribution of HLA among 31 MPM patients originating from the fibrous zeolite (erionite) and asbestos villages in central Anatolia, and two sets of controls. The cases represented all of the MPM cases diagnosed between 1995 and 1997 in our clinic at the Hacettepe University Hospital. One control group of 119 healthy individuals was drawn from Tuzkoy, which has the largest population of three erionite villages, a very high prevalence of mesothelioma due to environmental exposure to erionite, and accounted for 16 of the MPM cases in the study. A second control group composed of 118 renal transplant donors was formed for external comparison. RESULTS: A significant relation was found with the HLA-B41 antigen in 19.4% of the patients compared to 0.8% of the Tuzkoy inhabitants (odds ratio [OR] 28.3; 95% confidence interval [CI] 3.1-652.5) and 1.7% of the referent renal donor population (OR 13.9; 95% CI 2.3-106.7). The frequency of the HLA-B58 and -DR16 antigens was also observed to be significantly higher in patients with MPM compared to the two control groups. The odds ratios of MPM in those with HLA-B58 were 8.6 (95% CI 1.2-72.4) and 8.5 (95% CI 1.2 71.8), respectively, compared to those of the Tuzkoy inhabitants and renal donors. CONCLUSION: The predictive role of the HLA antigens -B41, -B58 and -DR16 for MPM needs to be further investigated. This will help in screening the population at risk, and facilitate preventive measures such as family counselling and gene therapy. PMID- 9869120 TI - Symptoms related to the sick building syndrome in a general population sample: associations with atopy, bronchial hyper-responsiveness and anxiety. AB - OBJECTIVE: The sick building syndrome (SBS) is a term mainly applied to irritative symptoms in the eyes, skin and upper airways that are experienced in certain environments. The aim of this study was to investigate the characteristics of subjects from the general population that report SBS-related symptoms. METHODS: The study comprised a randomly selected population of 418 individuals aged 20-45 years. The subjects underwent a structured interview, skin prick tests, a methacholine provocation test and answered two questionnaires respectively assessing psychiatric status and the presence of SBS symptoms. RESULTS: Eighty-seven subjects (21%) reported one or more SBS symptom weekly. Most common were general symptoms reported by 10% of the population. Female sex (odds ratio [OR] = 2.1), atopy (OR = 2.2) and anxiety (OR = 2.6) were significant independent risk factors for reporting at least one SBS-related symptom. Anxiety (OR = 3.2) and depression (OR = 4.4) were significant predictors for general symptoms, while bronchial hyper-responsiveness was a significant predictor for pharyngeal symptoms (dryness in the throat, sore throat and irritating cough) (OR = 5.4). CONCLUSIONS: Sick building symptoms are common in the general population and among women, while atopy and anxiety increase the risk of reporting such symptoms. Psychological factors are mainly related to general symptoms, while bronchial hyperresponsiveness is connected with pharyngeal symptoms. PMID- 9869121 TI - The association between epidemiological measures of the occurrence of asthma. AB - SETTING: The prevalence of asthma is useful for studying the causes of asthma. OBJECTIVE: To ascertain whether there is a relationship between the prevalence and incidence of asthma. DESIGN: The association between age and the occurrence of asthma was analysed in an epidemiological study of 15,813 persons. RESULTS: Different conclusions were reached depending on whether the point prevalence, cumulative prevalence or the incidence rates were studied. The relation between the incidence and prevalence of asthma is described in two epidemiological models, and none of the models seem to fit empirical data. Furthermore, it is shown that estimating incidence rates by prospectively measuring the point prevalences may introduce a considerable bias if the reliability of the diagnosis of asthma is around or below an agreement of 99%, which is probably usually the case. Including asthmatic symptoms during the last year in the definition of point prevalence means that there is no simple relation between incidence rates and point prevalences. CONCLUSION: The point prevalence may be a biased measure in the study of the etiology of asthma, as there is no simple relationship between the incidence and prevalence of asthma. PMID- 9869122 TI - Pulmonary reactions after exposure to 3-methylfuran vapour, a fungal metabolite. AB - A case of obstructive pulmonary reaction with flu-like symptoms after exposure to 3-methylfuran is described. This compound is produced by fungi, and can be found in buildings with mould growth. Previous studies have shown that exposure to the substance might increase the prevalence of respiratory symptoms, and is pneumotoxic to animals at high concentrations. PMID- 9869123 TI - Tuberculosis meningitis in Hong Kong: experience in a regional hospital. AB - Tuberculous meningitis (TBM) remains common in Hong Kong. From January 1996 to June 1997, 11 adult patients with TBM presented to Queen Mary Hospital, a regional hospital in Hong Kong. The annual incidence of TBM was estimated at 1.8 per 100,000 population. Nine patients were local Chinese, and only one patient had the acquired immune-deficiency syndrome (AIDS). In contrast to the classical presentation as a chronic indolent disease, our patients presented acutely: the mean duration from onset of symptoms to presentation was 4.8 days (range 0-10). The most common presenting symptoms were headache (64%), fever (46%), or both (36%), with focal deficits occurring in 64% of patients. Cerebrospinal fluid (CSF) culture and polymerase chain reaction (PCR) were positive in 30% and 29% of cases. Mean CSF cell count, protein and glucose levels were 340 x 10(6)/L, 267 mg/dL, and 2.3 mmol/L, respectively. Extra-neural tuberculosis occurred in 46% of cases. All patients survived and responded to treatment. Drug-induced hepatotoxicity was common; 64% of patients developed biochemical hepatitis. PMID- 9869124 TI - The significance of health sector reform for lung health services. International Union Against Tuberculosis and Lung Disease (IUATLD), and the International Development Research Centre (IDRC). Paris. 7-8 December 1997. PMID- 9869125 TI - Priorities for research in lung health. International Union Against Tuberculosis and Lung Disease (IUATLD), and the International Development Research Centre (IDRC). Paris. 9-11 December 1997. PMID- 9869126 TI - Tuberculosis and Horner's syndrome. PMID- 9869127 TI - High death rates for tuberculosis patients in rural South Africa. PMID- 9869128 TI - Fluoroquinolones and tuberculosis. PMID- 9869129 TI - The life span of silicone gel breast implants and a comparison of mammography, ultrasonography, and magnetic resonance imaging in detecting implant rupture: a meta-analysis. AB - Because of the growing concern surrounding the integrity and life span of silicone gel breast implants and the reported variations in the diagnostic accuracy of various imaging techniques in identifying ruptured implants, the authors undertook a meta-analysis of articles in the scientific literature to examine these concerns. They were able to include reports from the literature that detailed the condition and removal of 1,099 breast implants during the past 7 years. The median life span of a silicone gel implant was estimated to be 16.4 years. Of the implants, 79.1% were intact at 10 years, falling to 48.7% by 15 years. The sensitivities and specificities of three imaging modalities used in the diagnosis of implant rupture (mammography, ultrasonography, and magnetic resonance imaging [MRI]) were also evaluated and compared statistically in an effort to discover which of the three techniques might serve as the most reliable screening tool in the diagnosis of gel implant rupture. The sensitivity of mammography for finding a ruptured implant is 28.4% with a specificity of 92.9%. Ultrasonography has a sensitivity and specificity of 59.0% and 76.8% respectively compared with MRI, which was 78.1% and 80.0% respectively. For implants in place for 10 years, one would need to image 3.3 implants by ultrasound to identify a single possible rupture. However, because of the 76.8% specificity, 8.1 implants would need to be imaged to find a confirmed intraoperative rupture. This was similar to MRI, in which 3.1 implants would need to be imaged to detect one suspected rupture, and 6.1 implants would need to be imaged to find one intraoperatively confirmed rupture. The authors do not recommend either ultrasound or MRI as a screening tool based on their meta-analysis. PMID- 9869130 TI - Repair of unilateral cleft lip: a comparison of five techniques. AB - Repair of unilateral cleft lip is a challenging procedure with no single technique satisfactory for all types of unilateral cleft deformity. This study compares retrospectively five techniques of unilateral cleft lip repair in 72 children (45 boys, 27 girls). Twenty-two children had a Millard rotation advancement repair, 5 children had a Davies Z-plasty, 22 had a modified Z-plasty, 10 had a Tennison-Randall triangular flap repair, and 13 had a Nakajima-Yoshimura straight-line repair. Assessment was performed clinically by a plastic surgeon not involved in the original surgery. Repairs were assessed objectively by measurement of the vertical length of both the repaired and normal sides of the lip with calipers. Subjective criteria used to evaluate the repair were the symmetry of Cupid's bow, the quality of scar, the alignment of white roll, the evenness of the vermilion, and the lip pout. The mean follow-up period was 5.4 years (range, 9 months-29 years). Sixty-five of the 72 repairs measured (90%) were within two standard deviations (SDs) of normal and thus were considered to be of "acceptable" length. Seven repairs were unacceptably short (>2 SDs) on measurement, six of which included a rotation-advancement repair for a complete cleft lip. Subjective results paralleled the objective results. The outcome following repair of unilateral cleft lip was similar for all five surgical methods assessed except for complete cleft lips repaired by the rotation advancement technique, which tended to result in an unacceptably short lip as measured on the repaired side. PMID- 9869131 TI - An endoscopic subcutaneous dissector for obtaining vein grafts. AB - Recent improvements in internal mechanical retractors have resulted in a commercially available endoscopic subcutaneous dissector that has an inverted pistol grip for ease of retraction, an end-mounted endoscope channel, and a distal, translucent spoon-shaped shield that maintains the optical cavity. This provides an effective reach that is valuable for the minimally invasive harvest of any long, narrow structure--the greater saphenous vein in particular--as shown in this preliminary study with 2 patients. Such extracted vein grafts taken grossly from above and below the knee appear normal. Long-term histological changes and rates of conduit patency are still unknown. The limited-access incisions necessary for this endoscopic vein harvest definitely are smaller, and can be oriented transversely to minimize further the disfigurement from any resulting scar when compared with traditional methods. PMID- 9869132 TI - Endoscopic-assisted temporoparietal fascial flap dissection and harvesting: a feasibility preliminary cadaveric study. AB - Endoscopic procedures in plastic surgery have been applied in various aesthetic and reconstructive surgical techniques. The authors describe, in this preliminary report, a new surgical technique of endoscopic dissection of the temporoparietal fascial flap. A series of 6 fresh human cadavers (12 flaps) were dissected endoscopically. The surgical incisions, flap anatomy applicable to endoscopy, endoscopic surgical technique, and type of endoscopic setup is standardized for all flaps, allowing direct identification of the temporoparietal fascial layers and the major vascular pedicle. This endoscopic manipulation of the flap, without the traditional large scalp incision, permits local and free transfer of the temporoparietal fascial flap. Exposure of the flap by means of the conventional T or Y temporal incisions has several possible disadvantages, including an increased risk of blood loss, alopecia, and facial nerve injury. Endoscopic dissection and mobilization of the temporoparietal fascial flap can obviate the direct flap incision and yield a flap accessible to dissection and mobilization for additional clinical applications. This new, innovative, and minimally invasive endoscopic procedure may prove particularly applicable to future clinical applications of this type of fascial flap. PMID- 9869133 TI - Abdominal wall closure after selective aponeurotic incision and undermining. AB - The tension required to pull the anterior and the posterior rectus sheaths toward the midline was studied in 20 fresh cadavers at two levels: 3 cm above and 2 cm below the umbilicus. The quotient of the force used to mobilize the aponeurotic site to the midline and its resulting displacement was called the traction index. These indices were compared in three situations: (1) prior to any aponeurotic undermining, (2) after the incision of the anterior rectus sheath and the undermining of the rectus muscle from its posterior sheath, and (3) after additionally releasing and undermining the external oblique muscle. A significant decrease in aponeurotic resistance was observed after each dissection. The anterior sheath showed higher resistance to traction compared with the posterior sheath on both levels. No statistical difference was noted in the comparison of the values of the aponeurosis above and below the umbilicus. These results suggest that these procedures are effective in assisting in the closure of abdominal wall defects because these maneuvers decrease substantially the tension required for advancement of the aponeurotic edges. PMID- 9869134 TI - Diagnosis and treatment of retrobulbar hematoma resulting from blunt periorbital trauma. AB - Retrobulbar hematoma is a rare complication of blunt periorbital trauma with the potential disastrous consequence of visual impairment and blindness. The preoperative assessment, diagnostic symptoms and signs, as well as the treatment of this condition are reviewed. The patients presented as well as a review of the literature confirm that although retrobulbar hematoma is a rare complication of blunt periorbital trauma, irreversible visual sequelae can be prevented by prompt diagnosis and immediate surgical and pharmacological therapy. PMID- 9869135 TI - Intraoperative vascular monitoring of ipsilateral vs. contralateral TRAM flaps. AB - Transverse rectus abdominis musculocutaneous (TRAM) flap breast reconstruction has become an increasingly common procedure in the 1990s. The original description of the procedure was that of an ipsilaterally based pedicled procedure. However, because of concerns about potential folding of the pedicle with possible compromise of the vascular supply, with full muscle harvest the contralateral flap soon became the flap of choice. Subsequently there have been several large clinical series of pedicled TRAM flaps reported showing a relatively high complication rate related to problems with flap vascularity. These findings resulted in many centers favoring free TRAM flap breast reconstruction, despite an increased resource utilization and negligible differences in complication rates. Ipsilateral pedicled TRAM flap breast reconstruction is not a commonly reported procedure, and is reserved for patients in whom scars preclude use of the contralateral pedicle. The ease of flap shaping and lack of tension on the pedicle is recognized by some, and the ipsilateral procedure has become the flap of choice for these surgeons. The authors hypothesized that ipsilateral TRAM flaps have less compromise of venous outflow as a result of the rotational tension effect of transposing the flap to the mastectomy site. To determine if the vascularity of the pedicled TRAM procedure is influenced by pedicle choice, an intraoperative clinical study was designed to compare intravascular pressures in the deep inferior epigastric artery and vein pedicle at various times from pedicle division to flap insetting. Ten consecutive ipsilateral TRAM flaps were compared with eight contralateral flaps. The deep inferior epigastric pedicle was cannulated with both an arterial and venous catheter, and pressures were monitored from the time of division of the inferior pedicle through to the time of flap insetting. The venous pressures of the contralateral flaps were significantly higher at flap rotation and flap insetting compared with the ipsilateral group (p = 0.014 and p = 0.022 respectively). Perfusion pressures showed a greater decrease in the contralateral group from the time of pedicle division to the point of flap rotation and flap insetting compared with the ipsilateral group (p = 0.081 and p = 0.055). The authors conclude that pedicle tension in contralateral TRAM flaps is more common and results in greater venous resistance and decreased perfusion pressures. Versatility in flap shaping, improved maintenance of the IMF, and lack of disruption of the natural xyphoid hollow gives ipsilateral TRAM flaps additional advantages. PMID- 9869136 TI - Improving the donor site cosmesis of the latissimus dorsi flap. AB - A modification in the design of the pedicled latissimus dorsi musculocutaneous flap is described that aims to minimize the cosmetic morbidity of its donor site. The implications of this variation are discussed with particular reference to use of the flap in postmastectomy reconstruction. PMID- 9869137 TI - Reconstruction of lateral skull base oncological defects: the role of free tissue transfer. AB - Surgical ablation for oncological disease of the lateral skull base can result in extensive defects, with exposed bone and dura. Inadequate coverage can result in meningitis, osteomyelitis, or delay in adjuvant therapy. Successful reconstruction requires well-vascularized soft tissue and often a large cutaneous component. This study evaluates the role of free tissue transfer in reconstruction of lateral skull base defects. This study is a retrospective review of all patients undergoing lateral skull base resection for oncological disease and immediate reconstruction from 1993 through 1997. There were 18 patients with a mean age of 57 years. The temporal bone was resected in 50% of patients. All defects were reconstructed with free tissue transfers from the following donor sites: rectus abdominis (N = 14), latissimus dorsi (N = 2), anterolateral thigh (N = 1), and lateral arm (N = 1). A cutaneous skin island was employed in all patients. Free flap survival was 100%. Flap-related complications occurred in 33% of patients but did not delay the onset of adjuvant therapy. Vein grafts were not required to lengthen the vascular pedicle. Two patients required split-thickness skin grafts because of inadequate size of the skin island. Four patients underwent flap revision for contour deformity a mean of 4 months postoperatively. Free tissue transfer is a highly reliable method of reconstructing lateral skull base defects in a single stage. Careful flap selection and design can minimize the need for skin and vein grafts. The rectus abdominis donor site is preferred because of its location, large skin island, and excellent vascular pedicle. PMID- 9869138 TI - Muscle transposition flap for salvage of exposed peripheral prosthetic vascular grafts. AB - Infection of prosthetic vascular grafts, although infrequent, is a dreadful complication in vascular surgery. It is a challenge for the surgeon and is life- and limb-threatening to the patient. Conventionally, infection involving a foreign body is eradicated by removing the foreign body. Despite its theoretical appeal, this solution is fraught with danger--a high mortality rate that ranges between 10% and 30%, a high rate of protracted postoperative morbidity, and a limb amputation rate as high as 70%. The salvage of infected prosthetic vascular grafts by prompt soft-tissue coverage with transposition muscle flaps in 3 consecutive patients treated during a period of 12 years is the subject of this report. PMID- 9869139 TI - Hemoglobin oxygenation of venous-perfused forearm flaps. AB - To understand how venous flaps function we investigated whether blood flowing via the venous network reaches the capillaries of the skin. While measuring spectrophotometrically intracapillary hemoglobin oxygenation of fasciocutaneous forearm flaps in 12 patients, flap perfusion was changed by manipulating nutrient vessels. Conventionally perfused radial forearm flaps had an intracapillary hemoglobin oxygenation of 51% to 74% but decreased to 6.9% to 12.2% within 90 to 120 minutes after arterial occlusion and perfusion only from the cephalic vein entering the flap cranially (type I venous flap). Radial forearm flaps without any vascular connection showed no oxygenated hemoglobin after 180 to 240 minutes in the capillary network. After microsurgical vein anastomosis and release of the blood flow only via the cephalic or accompanying veins, hemoglobin oxygenation returned immediately to about 10%. We conclude from our results that there is actual capillary perfusion, albeit very slight, in type I venous forearm island flaps. PMID- 9869140 TI - Long-term follow-up of a patient with Romberg's disease reconstructed with free groin flaps. AB - Sixteen years ago the result of an operation that was performed on a 5-year-old boy with Romberg's disease was described in this journal. Nineteen years have passed since the first operation, and the authors now report the long-term follow up. The volume of the transferred free groin flaps was maintained. Although these flaps covered the patient's atrophic tissues as vascularized tissues, these flaps could not normalize the atrophy of the surrounding tissues, especially the bone. As for the orbital content, bony atrophy was more remarkable than the other tissues. PMID- 9869141 TI - Transconjunctival herniation of orbital fat. AB - The authors present 3 patients with subconjunctival fat prolapse treated at their oculoplastic unit. Albeit rare, orbital fat is a well-recognized entity, and is described in the literature as being associated with trauma and surgery. The 3 patients reported herein, however, presented with no history of trauma or surgery. This condition is produced by herniation of the intraconal fat between the conjunctiva and the sclera, presumably due to dehiscence of the Tenon's capsule. Differential diagnosis should be made with lacrimal gland ptosis, lacrimal gland tumors, and lymphoid tumors. PMID- 9869142 TI - Successful replantation of an avulsed middle finger. AB - In avulsion amputations of the digits, soft-tissue injuries are extensive and often require tendon, nerve, and vessel transfers or grafts. The functional results of such digital replantations are frequently less than ideal. Therefore, avulsion amputation of a single digit proximal to the insertion of the flexor digitorum superficialis has been a contraindication to replantation, because the anticipated poor functional result may interfere with overall hand function, and is not worth the sacrifice of a tendon, nerve, or vessel from another digit or transfer. The authors report a patient with avulsion amputation of the middle finger at the proximal interphalangeal joint. The digit was replanted successfully without any tissue transfers other than a radial digital artery from the ring finger. The functional results were good, and the authors believe that good functional and cosmetic results can be achieved in select patients with isolated digital avulsions, provided that an experienced hand microsurgeon and a skillful hand therapist are available for a compliant patient. PMID- 9869143 TI - Postoperative ramifications of total ear replantation. AB - Complete avulsion of the external ear, much less common than other minor ear trauma, presents a complex and difficult problem. The authors present a 35-year old male cigarette smoker who suffered traumatic avulsion of the right ear, and describe the replantation procedure and the patient's postoperative course. Aesthetic positioning of the replanted ear, treating the external auditory meatus, protecting the ear against exposure to the sun, and counseling the patient about issues related to sensation are important factors in the care of patients who have undergone such a procedure. PMID- 9869144 TI - Benign symmetric lipomatosis (Madelung's disease). AB - Benign symmetric lipomatosis, also known as Madelung's disease, is a rare condition characterized by massive fatty deposits arranged symmetrically around the neck, shoulders, and arms. These patients might present for liposuction and body contouring. Although infrequently encountered in the average plastic surgeon's practice, this condition should be considered when evaluating candidates for these procedures. The deformity is associated with chronic alcohol use and also with malignant tumors of the upper airways. The deformity is prone to recurrence and its surgical treatment often results in less than optimal aesthetic outcomes. Despite this fact, surgical removal via either direct excision or suction-assisted lipectomy provides the only real hope of palliation. This report describes a patient with this deformity and a review of the literature. PMID- 9869145 TI - Malignant blue nevus: three new cases and a review of the literature. AB - Three patients with malignant blue nevus are reported-one on the right side of the body, one on the right arm, and one on the face. The criteria and difficulty of histopathological diagnosis are considered as well as the differential diagnoses for this tumor. The therapy is described, and the possible relations between malignant blue nevus and certain other tumors (e.g., atypical or locally aggressive cellular blue nevus) are explored. A review of the literature reveals that there is current disagreement about the exact prognosis for these tumors and indicates the need to collect data for all patients observed. PMID- 9869146 TI - The present status of endoscopy. AB - Introduced to the field of plastic surgery in the early 1990s, surgical endoscopy has expanded rapidly with applications across the broad range of plastic surgery. Although basic principles and concepts have remained the same, there have been advances in both instrumentation and surgical ingenuity, allowing these broader applications. The numerous applications of the endoscope were reviewed and evaluated. Although some applications such as the endoscopic forehead lift and transaxillary endoscopic augmentation seem to have received general acceptance and are used across a broad spectrum of plastic surgery practices, others such as endoscopic muscle harvest have not gained such widespread acceptance. Also, there are areas such as endoscopic microvascular surgery that await advances in technology and instrumentation. PMID- 9869147 TI - It's the economy, stupid. PMID- 9869148 TI - Re: Treatment of scars: a review. PMID- 9869149 TI - Re: Aesthetic labia minora reduction. PMID- 9869150 TI - The use of a subatmospheric pressure dressing to salvage a Gustilo grade IIIB open tibial fracture with concomitant osteomyelitis to avert a free flap. PMID- 9869151 TI - Deletion polymorphism in the angiotensin-converting enzyme gene as a thrombophilic risk factor after hip arthroplasty. AB - Despite thromboprophylaxis, deep vein thrombosis is a common complication of major orthopedic surgery. Predisposing genetic risk factors are unknown. In this case-control study, we investigated the association of the insertion (I)/deletion (D) angiotensin converting enzyme (ACE) gene polymorphism, Factor V Leiden (R506Q) mutation, and 5,10 methylenetetrahydrofolate reductase (MTHFR) gene polymorphism with post-operative venous thrombosis in 85 patients who underwent elective total hip arthroplasty. The odds of a thrombotic event following hip surgery among subjects with the DD genotype of the ACE gene was increased more than 10-fold compared to subjects with the II genotype (odds ratio 11.7 [95% confidence interval 2.3-84.5]); it was increased 5-fold in subjects with the ID genotype compared to the II genotype (odds ratio 5.0 [95% confidence interval 1.1 34.9]). Mean plasma ACE level in control subjects not on ACE inhibitors at the time of study (n=43) was lowest in persons homozygous for the I allele (18.9+/ 7.95 U/l), intermediate in patients with the ID genotype (31.6+/-10.8 U/l) and highest in subjects homozygous for the D allele (44.0+/-7.14 U/l). Mean plasma ACE level among cases was higher (33.0 U/l, n=25) than among controls (29.4 U/l, n=43) but this difference was not statistically significant. Neither the Factor V Leiden mutation nor MTHFR gene polymorphism increased the risk of thrombosis following hip replacement. These results demonstrate that the I/D ACE gene polymorphism is a potent risk factor for thrombosis in subjects undergoing total hip arthroplasty. PMID- 9869152 TI - Hyperhomocysteinemia and venous thrombosis: a meta-analysis. AB - Hyperhomocysteinemia is an established risk factor for atherosclerosis and vascular disease. Until the early nineties the relationship with venous thrombosis was controversial. At this moment ten case-control studies on venous thrombosis are published. We performed a metaanalysis of these reports. We performed a MEDLINE-search from 1984 through June 1997 on the keywords "homocysteine" or "hyperhomocysteinemia" and "venous thrombosis", which yielded ten eligible case-control studies. We found a pooled estimate of the odds ratio of 2.5 (95% CI 1.8-3.5) for a fasting plasma homocysteine concentration above the 95th percentile or mean plus two standard deviations calculated from the distribution of the respective control groups. For the post-methionine increase in homocysteine concentration we found a pooled estimate of 2.6 (95% CI 1.6-4.4). These data from case-control studies support hyperhomocysteinemia as a risk factor for venous thrombosis. Further research should focus on the pathophysiology of this relationship and on the clinical effects of reducing homocysteine levels by vitamin supplementation. PMID- 9869153 TI - No association between the 20210 G/A prothrombin gene mutation and premature coronary artery disease. AB - The 20210 G/A prothrombin gene mutation is associated with an increased risk of venous thrombosis but whether there is an association of the mutation with premature coronary artery disease and acute myocardial infarction remains unclear. To further assess the role of the G/A genotype as a risk factor for arterial vascular disease, we performed a case-control study of 644 patients aged less than 50 years with angiographically proven coronary artery disease, 402 of whom had myocardial infarction, and 679 unrelated healthy control subjects aged less than 50 years, randomly selected from the electoral roll. The prevalence of the G/A genotype was 2.5% in patients with coronary artery disease, and 3.2% in control subjects (odds ratio 0.8; 95% confidence interval 0.35 to 1.83). The mutation was not more frequent among patients with a history of myocardial infarction (2.2%, odds ratio 0.7; 95% confidence interval 0.27 to 2.05), and there was no evidence of an interaction between the prothrombin mutation and conventional cardiovascular disease risk factors. There was no association between genotype and extent of angiographic coronary artery disease (p=0.73). We conclude that the 20210 G/A prothrombin gene mutation is not a major risk factor for premature coronary artery disease in our predominantly Caucasian Australian population. PMID- 9869154 TI - Augmented platelet aggregation as predictor of reocclusion after thrombolysis in acute myocardial infarction. AB - RATIONALE: Reocclusion after thrombolysis diminishes the benefits of early reperfusion after acute myocardial infarction (AMI). No clinical or laboratory variables have been identified as predictors for reocclusion yet. METHODS AND RESULTS: To evaluate hemostatic variables as potential risk determinants platelet aggregation (PA, representing platelet activity), thrombin/antithrombin complexes (TAT, representing thrombin generation), and plasminogen activator inhibitor type 1 (PAI-1, representing endogenous fibrinolysis) were determined in 31 patients with AMI at 0, 1, 2. and 12 h after the start of thrombolysis as well as at hospital discharge. Reocclusion (defined as reinfarction or angiographically confirmed, clinically silent coronary reocclusion) occurred in 5 patients within 5-14 days and in 8 patients within 1 year. TAT plasma concentrations were lower in patients with reocclusion than in those without (9.9+/-5.7 vs. 22.9+/-22.2 ng/ml at 2 h, 6.5+/-3.1 vs. 1 1.2+/-6.4 ng/ml at 12 h, means+/-SD, p <0.05 each). Neither concentration nor activity of PAI-1 in plasma differed between both patient groups. However, both slope and maximum of PA (induced by 2 micromol/l ADP) were augmented in patients with reocclusion (slope: 39.4+/-1.7 vs. 32.5+/ 7.4 at 2 h, p <0.001; 42.6+/-2.6 vs. 36.6+/-8.9 at 12 h, p <0.01). Results were independent of the thrombolytic agent used (alteplase or reteplase). A PA slope at 2 h higher than the average slope before thrombolysis (37.2+/-5.7) could be identified as best predictor for early (within 5-14 d, p=0.017, sensitivity 1.00, specificity 0.69) and late reocclusion (within 1 y, p=0.009, 0.88 and 0.74, respectively). CONCLUSIONS: Increased PA following coronary thrombolysis appears to be associated with early and late reocclusion. This marker could be useful in identifying patients who may benefit from more aggressive antiplatelet (such as GP IIb/IIIa receptor antagonists), interventional, or both strategies. PMID- 9869155 TI - A cost-effectiveness analysis of aspirin versus oral anticoagulants after acute myocardial infarction in Italy -- equivalence of costs as a possible case for oral anticoagulants. AB - AIMS: The recent publication of two large trials of secondary prevention of coronary artery disease with oral anticoagulants (WARIS and ASPECT) has caused a revival of the interest for this antithrombotic therapy in a clinical setting where the use of aspirin is common medical practice. Despite this, the preferential use of aspirin has been supported by an American cost-effectiveness analysis (JAMA 1995; 273: 965). METHODS AND RESULTS: Using the same parameters used in that analysis and incidence of events from the Antiplatelet Trialists Collaboration and the ASPECT study, we re-evaluated the economic odds in favor of aspirin or oral anticoagulants in the Italian Health System, which differs significantly in cost allocation from the United States system and is, conversely, similar to other European settings. Recalculated costs associated with each therapy were 2,150 ECU/ patient/year for oral anticoagulants and 2,187 ECU/patient/year for aspirin. In our analysis, the higher costs of oral anticoagulants versus aspirin due to a moderate excess of bleeding (about 10 ECU/ patient/year) and the monitoring of therapy (168 ECU/ patient/year) are more than offset by an alleged savings for recurrent ischemic syndromes and interventional procedures (249 ECU/ patient/year). CONCLUSIONS: Preference of aspirin vs. oral anticoagulants in a pharmaco-economical perspective is highly dependent on the geographical situation whereupon calculations are based. On a pure cost effectiveness basis, and in the absence of data of direct comparisons between aspirin alone versus I.N.R.-adjusted oral anticoagulants, the latter are not more expensive than aspirin in Italy and, by cost comparisons, in other European countries in the setting of post-myocardial infarction. PMID- 9869156 TI - Tissue factor expression and metastatic potential of colorectal cancer. AB - Several studies have previously demonstrated tissue factor (TF) expression in solid tumors. In our study, we evaluated by immunohistochemical staining TF expression in 79 cases of colorectal cancer and 17 cases of metastatic cancer of the liver from colorectal cancer, and investigated the relationship between the clinicopathological features and TF expression. TF was detected in the tumor of 57% of colorectal cancer patients, and its expression was significantly increased (p=0.01) in metastatic tumors (88%). TF expression was more commonly observed in metastatic tumors than in any Dukes' stage of primary cancer. In primary cancer, the detection of TF was more frequent in cases with lymph node metastasis (Dukes' C, 63%) or with hematogenous metastasis (Dukes' D, 82%) than in tumors without lymph node or hematogenous metastasis (Dukes' A and B, 46%, p=0.03). These results suggest that the expression of TF is related with the metastatic potential of colorectal cancer. PMID- 9869157 TI - Warfarin or acenocoumarol: which is better in the management of oral anticoagulants? AB - Warfarin is employed more frequently than acenocoumarol because of its longer half-life (36 h), theoretically providing more stable anticoagulation, and avoiding factor VII fluctuations that potentially occur during acenocoumarol treatment (half-life 10 h). The aim of our study was to compare acenocoumarol with warfarin in the same group of 103 patients who started oral anticoagulation with acenocoumarol and then changed to warfarin. In these patients we compared the previous period of six months on acenocoumarol treatment (July-December 1996) with a new six-month period on warfarin (July-December 1997). We wished to know whether warfarin could improve the quality and the stability of oral anticoagulation of our patients and whether there was a difference between the two drugs in the weekly mean dose per patient. Moreover in order to detect the possible daily fluctuation of factor VII, we evaluated a further group of 54 patients. A subgroup of these patients was treated with warfarin while another received acenocoumarol. In the first group of patients, 1,158 and 1,064 PTs were carried out with acenocoumarol and warfarin, respectively. The percentage of PTs in the therapeutic range was 59% with acenocoumarol and 62% with warfarin (p=0.4). The mean number of visits per patient was 12 and 11, and the mean number of visits in the therapeutic range was 7 and 7, respectively. The last check in file method did not show any difference between the two drugs. Overdose states were 51 (4.4%) with acenocoumarol and 30 (2.8%) with warfarin (p=0.4). A good correlation (r=0.92) was found between the acenocoumarol and the warfarin weekly mean dose. The mean warfarin/acenocoumarol weekly dose ratio was 2.08 (range: 1.25-3.30; CI 95%: 1.99-2.16). In the second group of patients, factor VII levels with both drugs were higher 24 h after administration than 16 h after, showing that their daily fluctuation was independent of the drug's half-life, since factor VII levels in patients with a low vitamin K intake were not increased. Our results showed that warfarin did not appear to be better than acenocoumarol in the performance of an Anticoagulation Clinic in terms of PTs within the therapeutic range per patient. It seems that the behaviour of factor VII was affected by the intake of vitamin K rather than by the short half-life of acenocoumarol. PMID- 9869158 TI - Contribution of acute-phase proteins and cardiovascular risk factors to erythrocyte aggregation in normolipidemic and hyperlipidemic individuals. AB - BACKGROUND: Numerous studies have demonstrated that elevated concentrations of acute-phase proteins affect red blood cell (RBC) aggregation. Plasma lipids and lipoproteins were also shown to be correlated with RBC aggregation in hypercholesterolemia. However, whether acute-phase proteins promote RBC hyperaggregation in hyperlipidemic patients is unknown. The main objective of the study was to identify the impact of acute-phase proteins such as fibrinogen (Fib), haptoglobin (Hp), ceruloplasmin (Cp), alpha1-acid glycoprotein (AGP), alpha1-antitrypsin (AT), immunoglobulin G (IgG), and albumin (Alb) on RBC aggregation in 35 hyperlipidemic patients. The influence of these proteins in 32 normolipidemic subjects was also determined. METHODS AND RESULTS: RBC aggregation parameters reflecting the kinetics of rouleau formation and the adhesive strength between RBCs were measured by laser reflectometry. Multivariate forward stepwise linear regression analyses were performed to study the relationship between RBC aggregation and these acute-phase proteins, total cholesterol (TC), triglycerides (TG), high (HDL-C) and low (LDL-C) density lipoprotein cholesterol, age, gender, body mass index (BMI), mean blood pressure (Mpresure), and smoking habit. The kinetics of rouleau formation was positively correlated with the linear combination of IgG and Hp (r=0.76, p <0.0001) in hyperlipidemic patients, whereas IgG, smoking, AGP and gender were significant independent predictors in healthy subjects (r=0.79, p <0.0001). The correlations obtained for the models predicting the adhesive strength between RBCs were 0.69 in patients (Alb, HDL-C, IgG, p <0.002) and 0.71 in healthy individuals (AGP, BMI, p <0.0001). CONCLUSION: This study suggests that acute-phase proteins such as IgG, Hp, AGP and Alb influence significantly and in an independent way the level of RBC aggregation. The close association between RBC aggregation and cardiovascular risk factors further strengthens its clinical importance. PMID- 9869159 TI - Retrospective neuropathological review of prion disease in UK haemophilic patients. AB - In 1996, the CJD surveillance unit in Edinburgh, UK described nvCJD which was thought to be the human equivalent of bovine spongiform encephalopathy (BSE). The identification of prion protein in the tonsil of an affected individual has raised the question of transmission of nvCJD via blood products. This study examines the post mortem brains of 33 patients who were treated with clotting factor concentrate of predominately UK donor source during the years 1962-1995. The brains were examined by conventional histological methods and also for the prion protein using monoclonal antibodies KG9 and 3F4. No evidence of spongiform encephalopathy was found and the immunocytochemistry was negative for PrP in all cases. It is concluded that, at present, there is no evidence for the transmission of nvCJD via clotting factor concentrate to patients with haemophilia. PMID- 9869160 TI - Home treatment of mild to moderate bleeding episodes using recombinant factor VIIa (Novoseven) in haemophiliacs with inhibitors. AB - OBJECTIVE: To assess the safety and efficacy of a fixed dose of recombinant activated factor VII (rFVIIa; NovoSeven) in the home setting for mild to moderately severe joint, muscle; and mucocutaneous bleeding episodes in patients with haemophilia A or B with inhibitors. DESIGN: Multicentre, open-label, single arm, phase III study of one year duration. METHODS; Patients or their caregivers administered up to three doses of rFVIIa (90 microg/kg i.v.) at 3 h intervals within 8 h of the onset of a mild to moderate bleeding episode. Once the subject considered that rFVIIa had been "effective" with regard to haemostasis (after 1-3 injections), one further (maintenance) dose of rFVIIa was administered. RESULTS: Of 60 patients enrolled, 56 experienced at least one bleed, and 46 completed the one year study. 614 of 877 bleeds (70%) were evaluable according to protocol definitions. Haemostasis was rated as "effective" in 92% (566/614) of evaluable bleeds after a mean of 2.2 injections. For successfully treated episodes, the time from onset of bleeding until administration of the first injection was 1.1+/ 2.0 h (mean+/-SD). Twenty-four hours after initial successful response, haemostasis was reported as having been maintained in 95% of cases. Efficacy was comparable for muscle, joint and target joint, and mucocutaneous bleeding episodes. In an intent-to-treat analysis of all 877 bleeding events, efficacy outcomes were equivalent to the evaluable bleeds, with an effective response in 88% of treated episodes. Treatment-related adverse events occurred in 32 (3% of all) bleeding episodes and consisted of re-bleeds/new bleeds in more than 50% (18/32) of these events. A single episode of superficial thrombophlebitis was the only thrombotic complication encountered, and there were no patient withdrawals due to adverse events. Development of FVII(a) antibodies could not be detected, and hypersensitivity reactions to rFVIIa were not reported. CONCLUSION: rFVIIa is effective and well tolerated when used in the home setting to treat mild to moderate bleeding episodes in patients with haemophilia A or B with inhibitors. PMID- 9869161 TI - A cross-over pharmacokinetic study of a double viral inactivated factor IX concentrate (15 nm filtration and SD) compared to a SD factor IX concentrate. AB - A double blind randomized cross-over multi-center study has been conducted to compare the pharmacokinetic and coagulation activation markers of high-purity factor IX concentrate subjected to both solvent/ detergent (SD) treatment and 15 nm-filtration (FIX-SD-15) with the licensed product subjected only to solvent detergent (FIX-SD). This filtration process allows the elimination of small particles, such as non-enveloped viruses (i.e., hepatitis A and parvovirus B19). Eleven severe hemophilia B patients (FIX coagulant activity <2 IU/dl) received one infusion of 60 IU/kg of FIX-SD and one infusion of 60 IU/kg of FIX-SD-15 at least at 10 days interval. Blood samples were obtained before and at various time up to 72 h after infusion. The decay curves of factor IX (FIX:C and FIX:Ag) were evaluated by a model independent method. Bioequivalence was found between the two concentrates using the Schuirmann test. The mean FIX:C and FIX:Ag recovery of FIX SD-15 was 1.08 and 0.89 IU/dl/IU/kg respectively with a mean half-life of 33.3 h for FIX:C and 25.6 h for FIX:Ag. Six months after initial enrollment, pharmacokinetic parameters were similar in the 7 patients tested. There was no significant variation of prothrombin fragment 1+2 and thrombin-antithrombin complexes measured up to 6 h after infusion, indicating that there was no activation process after administration of FIX. In conclusion, these data demonstrate that the introduction of a 15 nm filtration does not alter the pharmacokinetic profile of a well characterized SD FIX concentrate while providing additional viral safety. PMID- 9869163 TI - A plasma coagulation assay for an activated protein C-independent anticoagulant activity of protein S. AB - To study the physiological importance of the activated protein C (APC) independent anticoagulant activity of protein S, we developed an assay specific for this activity. The ability of protein S to prolong the clotting time in an APC-independent way was expressed as the ratio of the clotting time in a plasma sample divided by the clotting time in the presence of a polyclonal antibody against human protein S (both after 1:1 dilution in protein S-C4BP deficient plasma). The mean protein S-dependent anticoagulant ratio (PSdAR) was 1.53+/-0.18 in 34 healthy controls, and was significantly lower in 16 heterozygous protein S deficient patients (PSdAR=1.15+/-0.09). In plasmas from patients under oral anticoagulant therapy the mean PSdAR was within the range of the control population (1.50+/-0.18). The mean total protein S antigen level in these plasmas was 58%, suggesting a higher specific APC-independent anticoagulant activity of protein S in these patients than in normals. This functional protein S assay can be used for the laboratory diagnosis of protein S deficiency, and to study the mechanism of the APC-independent anticoagulant activity in plasma. PMID- 9869162 TI - Effects of recombinant human soluble thrombomodulin (rhs-TM) on clot-induced coagulation in human plasma. AB - Recent studies have suggested that clot-bound thrombin plays an important role in thrombus growth. In this study, we examined the effects of recombinant human soluble thrombomodulin (rhsTM) on clot-induced coagulation. rhsTM enhanced the activation of protein C by clots, and attenuated clot-induced thrombin generation and fibrinopeptide A (FPA) production in a dose-dependent manner. The inhibitory effect of rhsTM was abolished by anti-protein C antibody. The inhibitory effect of rhsTM on clot-induced thrombin generation continued for over 60 min after the addition of the clot, while an active site-directed thrombin inhibitor, argatroban, produced a more transient inhibition. rhsTM also inhibited the regrowth of the clot in (125)I-fibrinogen-supplemented plasma. We also examined the effect of rhsTM by thromboelastography, rhsTM reduced the growth of the clot but had little effect on the time to begin clotting, while heparin and Fragmin (low molecular weight heparin) had effects opposite to those of rhsTM. These findings suggest that rhs-TM attenuates the growth of the clot by activating protein C and inhibiting further thrombin generation in the clot. PMID- 9869164 TI - Inhibition of lupus anticoagulant activity by hexagonal phase phosphatidylethanolamine in the presence of prothrombin. AB - We have previously demonstrated that lupus anticoagulant antibodies from patients with systemic lupus erythematosus (SLE) specifically recognize hexagonal (II) phase phosphatidylethanolamine (PE), but not bilayer PE (Thromb Haemost 1989; 62: 892). In those studies, the involvement of proteins in this recognition was not evaluated. To address this issue, we have isolated IgG lupus anticoagulant antibodies from the plasma of SLE patients and evaluated the inhibition of lupus anticoagulant activity by hexagonal (II) phase PE in the presence and absence of purified plasma proteins. All six of the IgG lupus anticoagulant antibodies tested were inhibited by hexagonal (II) phase PE in the presence, but not the absence, of human prothrombin. In contrast, little or no inhibition was observed with prothrombin alone or with PE in combination with either beta2-glycoprotein I or annexin V. These data indicate that, for certain lupus anticoagulant antibodies, inhibition by hexagonal (II) phase PE is dependent on prothrombin, suggesting that these antibodies recognize a complex of PE and prothrombin. PMID- 9869165 TI - Fibrate-modulated expression of fibrinogen, plasminogen activator inhibitor-1 and apolipoprotein A-I in cultured cynomolgus monkey hepatocytes -- role of the peroxisome proliferator-activated receptor-alpha. AB - Fibrates are used to lower plasma triglycerides and cholesterol levels in hyperlipidemic patients. In addition, fibrates have been found to alter the plasma concentrations of fibrinogen, plasminogen activator inhibitor-1 (PAI-1) and apolipoprotein A-I (apo A-I). We have investigated the in vitro effects of fibrates on fibrinogen, PAI-1 and apo A-I synthesis and the underlying regulatory mechanisms in primary monkey hepatocytes. We show that fibrates time- and dose dependently increase fibrinogen and apo A-I expression and decrease PAI-1 expression in cultured cynomolgus monkey hepatocytes, the effects demonstrating different potency for different fibrates. After three consecutive periods of 24 h the most effective fibrate. ciprofibrate (at 1 mmol/l), increased fibrinogen and apo A-I synthesis to 356% and 322% of control levels, respectively. Maximum inhibition of PAI-1 synthesis was about 50% of control levels and was reached by 1 mmol/l gemfibrozil or ciprofibrate after 48 h. A ligand for the retinoid-X receptor (RXR), 9-cis retinoic acid, and specific activators of the peroxisome proliferator-activated receptor-alpha (PPARalpha), Wy14,643 and ETYA, influenced fibrinogen, PAI-1 and apo A-I expression in a similar fashion, suggesting a role for the PPARalpha/RXRalpha heterodimer in the regulation of these genes. When comparing the effects of the various compounds on PPARalpha transactivation activity as determined in a PPARalpha-sensitive reporter gene system and the ability of the compounds to affect fibrinogen, PAI-1 and apo A-I antigen production, a good correlation (r=0.80; p <0.01) between PPARalpha transactivation and fibrinogen expression was found. Apo A-I expression correlated only weakly with PPARalpha transactivation activity (r=0.47; p=0.24), whereas such a correlation was absent for PAI-1 (r=0.03; p=0.95). These results strongly suggest an involvement of PPARalpha in the regulation of fibrinogen gene expression. PMID- 9869166 TI - Identification and characterization of two thrombin-activatable fibrinolysis inhibitor isoforms. AB - Thrombin-activatable fibrinolysis inhibitor (TAFI) is synthesized by the liver and is thought to circulate in plasma as a plasminogen-bound zymogen. When it is activated by the thrombin/thrombomodulin complex, activated TAFI exhibits carboxypeptidase B-like activity. To study the structure-function relationship of TAFI, we expressed recombinant human TAFI in insect cells. During the cloning of TAFI cDNA from several human liver cDNA libraries, we identified a second TAFI cDNA which differed from the published sequence at 2 positions. One of these sequences resulted in a substitution of alanine for threonine at residue 147, the other was a silent mutation. These substitutions were found in several cDNA libraries from different sources. Using Southern blot analysis, we confirmed the existence of this TAFI polymorphism in the population. In order to compare the activation and activity of TAFI isoforms, we expressed both isoforms in the baculovirus expression system, and compared the enzyme kinetics of the purified proteins. The molecular weight of recombinant TAFI is lower than plasma TAFI due to differences in glycosylation. The two recombinant TAFI isoforms had similar activation kinetics and the activated enzymes had similar carboxypeptidase B-like activity towards small molecule substrates. Their ability to retard clot lysis was found to be similar in a plate clot lysis assay. PMID- 9869167 TI - 4G/5G polymorphism of PAI-1 gene promoter and fibrinolytic capacity in patients with deep vein thrombosis. AB - A deletion/insertion polymorphism (4G or 5G) in the promoter of the plasminogen activator inhibitor type 1 gene has been suggested to be involved in regulation of the synthesis of the inhibitor, the 4G allele being associated with enhanced gene expression. A relationship between 4G/5G polymorphism and PAI-1 levels was found in patients with cardiovascular and metabolic diseases, but not in healthy subjects. In the present work we studied the distribution of PAI-1 4G/5G genotype and its relation to fibrinolytic capacity in 70 unrelated patients with deep vein thrombosis. Each patient was assayed before and after 20 min. Venous occlusion for euglobulin lysis time, t-PA antigen and activity, and PAI-1 antigen and activity. The prevalence of 5G homozygous carriers was significantly lower in patients than in controls (10% vs. 26%, p=0.009). The 5G allele frequency was reduced, even though not significantly, in DVT patients compared to healthy subjects (0.40 vs. 0.51, respectively). In the patient group, the mean PAI-1 antigen and activity levels were significantly higher than among controls and related to the 4G/5G polymorphism. In patients with 4G/5G and 4G/4G genotype a significant correlation was found between PAI-1 levels and the global fibrinolytic activity as evaluated by euglobulin lysis time. The prevalence of a reduced fibrinolytic potential due to PAI-1 excess was 45.7% among DVT patients. Moreover, the prevalence of PAI-1 induced hypofibrinolysis was strongly related to PAI-1 polymorphism, since it was significantly lower in 5G homozygous patients (28.6%) than in both 4G/5G carriers (55.3%, p <0.001) and 4G homozygous patients (57.9%, p <0.001). In conclusion, in patients with deep vein thrombosis the 4G polymorphism of PAI-1 gene promoter may influence the expression of PAI-1 and it should be taken into consideration as a facilitating condition for pathological fibrinolysis together with other environmental and genetic factors. Whether this has any significance in regard to the pathogenesis of venous thrombosis remains to be proven. PMID- 9869168 TI - Evidence for the expression of urokinase-type plasminogen activator by human venous endothelial cells in vivo. AB - Endothelial cells (ECs) in culture synthesize and secrete urokinase-type plasminogen activator (u-PA), but the normal vascular endothelium is believed to synthesize only tissue plasminogen activator (t-PA), which is thought to be responsible for intravascular fibrinolysis. More recently, animal studies have shown that the biological role of u-PA in fibrinolysis has been underestimated, prompting a re-examination of its synthesis by the endothelium. In this study, we investigated whether u-PA was synthesized by non-atherosclerotic endothelial cells in vivo by testing ECs dislodged by venipuncture from 12 normal volunteers and 17 patients admitted for plasmapheresis. The ECs were isolated with an anti endothelial monoclonal antibody coupled to immunomagnetic beads and characterized by morphology and by labelling for vWF, CD31, and UEA-1 binding. U-PA antigen was found in 50% of the ECs from the normal subjects and in 60% of those from patients. U-PA enzymatic activity on zymograms was detected in 50% of the normal samples and 60% of the patient samples, with the latter being more frequently and more strongly positive. U-PA mRNA was found in all the normal and patient samples tested. The results indicate that u-PA is synthesized by the venous endothelium in vivo but that its expression is highly variable. PMID- 9869169 TI - The effects of Mpl-ligand, interleukin-6 and interleukin-11 on megakaryocyte and platelet alpha-granule proteins. AB - Thrombopoietin (Mpl ligand), interleukin-6 (IL-6), and interleukin-11 (IL-11) differ in their effects on megakaryocyte maturation and development. In the present study, the impact of these thrombocytopoietic cytokines on biochemical and structural granule and membrane components was examined. Western blotting was performed on equivalent amounts of isolated megakaryocyte or platelet protein and the relative intensities of the enhanced chemiluminescent-visualized bands were quantitated by densitometry. Megakaryocyte growth and development factor (MGDF), a recombinant thrombopoietin-related molecule, significantly increased megakaryocyte fibronectin (72%), thrombospondin (55%), von Willebrand factor (28%) and p-selectin (CD62p) (37%) when compared to equivalent amounts of protein from saline-treated controls (p<0.01). Megakaryocyte fibrinogen was not increased. Ultrastructurally, there was a marked increase in ribosomes and rough endoplasmic reticulum even in mature-appearing megakaryocytes. Platelets from MGDF-treated mice showed small increases in fibronectin (8%), and CD62p (18%), but did not show increases in other measured alpha-granule proteins. Neither IL-6 nor IL-11 increased megakaryocyte or platelet alpha-granule proteins over levels observed in saline controls. IL-11 treated megakaryocytes, while also exhibiting an increase in ribosomes, were characterized by prominent cytoplasmic fragmentation. The study demonstrates the impact of Mpl ligand in increasing synthesized megakaryocyte alpha-granule proteins and of IL-11 in promoting megakaryocyte fragmentation. PMID- 9869170 TI - Secreted ADP plays a central role in thrombin-induced phospholipase D activation in human platelets. AB - Thrombin and other agonists that induce secretion and aggregation in human platelets also activate phospholipase D (PLD), but the signaling cascade leading to activation of PLD in human platelets is not yet clear. We have determined that apyrase, which scavenges ADP secreted during platelet activation, is able to block or reduce the PLD activation stimulated by low (0.1 U/ml or less) or high (0.3- 1.0 U/ml) concentrations of thrombin, respectively. Neither ADP (up to 100 microM) nor its more potent analogue 2-methylthio-ADP (up to 100 microM), however, are able to stimulate PLD alone, and even the addition of fibrinogen, which results in platelet aggregation, is not sufficient for PLD activation. In contrast, ADP is able to stimulate PLD in the presence of low concentrations of thrombin that alone have little or no effect, suggesting ADP may play an amplifying role in platelet PLD activation. This hypothesis is supported by the finding that the purinergic receptor antagonist ARL 66096, an ATP analogue, reduces in a concentration-dependent fashion the PLD response to thrombin (IC50=28 nM with 0.1 U/ml thrombin). ARL 66096 also abolishes the PLD activation by ADP observed in the presence of low concentrations of thrombin, confirming that the antagonist inhibits an ADP-dependent component of the response. In addition, the thromboxane A2 receptor agonist U46619 activates PLD, and this response is markedly reduced by ARL 66096. Concomitantly, phosphorylation of the protein kinase C substrate pleckstrin in response to thrombin or U46619 is partially or totally inhibited by ARL 66096, respectively, consistent with ADP stimulation of protein kinase C being involved in the PLD response to these agonists. Based on these findings, we conclude that ADP secretion and activation of purinergic ADP receptors is an important amplification mechanism in the signal transduction pathways leading to PLD activation in human platelets. PMID- 9869171 TI - Interaction of leukocytes with platelet microparticles derived from outdated platelet concentrates. AB - Platelet microparticles (PMP) were isolated from outdated platelets by a combination of differential centrifugation and gel filtration, and the concentration of PMP was expressed in the equivalent of GPIIb/IIIa complex measured by captured ELISA. PMP bound to isolated neutrophils and macrophages in a dose-dependent manner, but they did not bind to lymphocytes. Incubation of PMP with neutrophils did not activate these cells as measured by up-regulation of Mac 1, release of human granulocyte elastase, and calcium mobilization. Incubation of PMP with macrophages did not enhance IL-8 production and the oxygen burst but slightly and significantly increased production of MCP-1. After 10 min incubation of PMP with macrophages, an increase of GPIIb/IIIa antigen was observed suggesting that PMP may be endocytosed by macrophages. In conclusion, PMP bind to leukocytes, but, in contrast to activated platelets, do not play a significant role in leukocyte activation. PMID- 9869172 TI - In vitro efficacy of platelet glycoprotein IIb/IIIa antagonist in blocking platelet function in plasma of patients with heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is an important complication following administration of heparin. Platelet activation and aggregation induced by heparin/platelet factor 4/immunoglobulin complexes are thought to be the underlying mechanism for this condition, so it was hypothesized that abciximab (a humanized murine monoclonal antibody directed against the glycoprotein IIb/IIIa receptor) would prevent heparin-induced platelet aggregation and activation in plasma from patients with HIT. Platelet aggregation was tested in vitro with platelet-poor plasma (obtained from 23 patients with HIT), platelet-rich plasma (from normal donors with known reactivity), heparin (0.5 U/ml), and ascending doses of abciximab (0.07-0.56 microg/ml). The ability of abciximab to prevent platelet activation was also evaluated using flow cytometry (P selectin expression, mepacrine release, microparticle formation) and platelet factor 4 immunoassay. In vitro, abciximab inhibited heparin-induced platelet aggregation in a dose-dependent fashion (IC50 0.103 microg/ml) and inhibited microparticle formation, the expression of P-selectin, release of mepacrine and platelet factor 4. These findings suggest that abciximab may be useful in treatment of patients with HIT and warrants further clinical evaluation. PMID- 9869173 TI - Effect of glycoprotein IIb-IIIa receptor antagonism on platelet membrane glycoproteins after coronary stent placement. AB - Platelet membrane glycoproteins play a crucial role in ischemic complications after coronary stenting. Glycoprotein IIb-IIIa blockade reduces adverse clinical events after angioplasty but is associated with rare but profound thrombocytopenia that might increase hemorrhagic complications. Changes in platelet membrane glycoproteins of patients with angina who underwent coronary stenting and were treated with the GPIIb-IIIa antagonist abciximab (n=20) or with heparin (n=23) were studied. GPIb-IIIa receptor blockade and membrane glycoproteins were evaluated with immunological markers in venous blood samples taken before. 10, 24, 48, 72, and 96 h after initial treatment with either abciximab or heparin. Patients receiving abciximab therapy showed a rapid inhibition of binding of fluorochrome-conjugated mAb CD41 and c7E3 concomitant with a reduction in platelet aggregation which was restored in part in the days after termination of abciximab infusion. Induction of ligand-induced binding sites on GPIIb-IIIa was increased in patients receiving abciximab. The expression of ligand-induced binding sites correlated inversely with platelet count. No significant change in platelet membrane markers were found in the heparin group. In vitro studies showed that abciximab induces ligand-induced binding sites on isolated platelets and on nuclear cells bearing recombinant GPIIb-IIIa. Abciximab rapidly achieves GPIIb-IIIa receptor blockade after coronary stent placement that might be beneficial in high-risk settings to bridge the delayed action of ticlopidine. Significant alterations of platelet membrane glycoproteins during GPIIb-IIIa antagonism might contribute to development of acute profound thrombocytopenia. PMID- 9869174 TI - von Willebrand factor (vWf) as a plasma marker of endothelial activation in diabetes: improved reliability with parallel determination of the vWf propeptide (vWf:AgII). AB - Elevated plasma von Willebrand factor (vWf) levels are found in diabetes and other vasculopathies, and predict cardiovascular mortality. vWf is stored and released from endothelial cell secretory granules, along with equimolar amounts of its propeptide (vWf:AgII). In the present study, we examined plasma propeptide levels as a marker of endothelial secretion in vivo, using an ELISA based on monoclonal antibodies. vWf but not propeptide levels are influenced by blood groups, explaining in part the smaller variation in plasma propeptide levels among normal individuals. In both controls and insulin-dependent diabetic patients, we found a close correlation between propeptide and immunoreactive vWf levels (r2=0.54, p <0.0001). vWf and propeptide were elevated in patient subgroups with microalbuminuria or overt diabetic nephropathy, whereas only the propeptide was significantly elevated in the normoalbuminuric subgroup. This observation suggests that in conjunction with vWf, propeptide measurements may improve the identification of endothelial activation, which occurs frequently even without increased urinary albumin excretion. In 12 NIDDM patients, a 3-week diet enriched in monounsaturated fat (MUFA) resulted in parallel decreases in vWf (-22%, p <0.05) and propeptide (-17%, p <0.05) levels, indicating that the experimental diet affected endothelial secretion rather than vWf catabolism. A carbohydrate-enriched control diet did not significantly influence either marker. Our results suggest that concomitant determinations of plasma vWf and propeptide are useful tools to assess endothelial activation in vivo, and reinforce our previous conclusion that a diet rich in MUFA can improve endothelial function in NIDDM. PMID- 9869175 TI - Activated platelets induce tissue factor expression on human umbilical vein endothelial cells by ligation of CD40. AB - CD40 is a type I member of the tumour necrosis factor (TNF) receptor superfamily of proteins, and is present on a wide variety of cells including vascular endothelial cells. Ligation of this receptor on endothelial cells is known to increase expression of inflammatory adhesion molecules. We have recently demonstrated that platelets express the ligand of CD40 (CD154) within seconds of exposure to agonist, and interact with endothelial cells to participate directly in the induction of an inflammatory response. Here we show that activated platelets induce tissue factor (TF) expression on endothelial cells in a CD40/CD154-dependent manner, and that the magnitude of this response can equal that induced by TNFe. Moreover, CD40 ligation on endothelial cells downregulates the expression of thrombomodulin. We also show that CD40-mediated TF expression is less sensitive to inhibition with the oxidative radical scavenger pyrrolidine dithiocarbamate than is that mediated by TNFalpha, indicating that CD40 has a distinct signalling pathway. Tissue factor is a cell membrane protein which functions as the main trigger of the extrinsic pathway of blood coagulation, and its expression on endothelial cells is implicated in wound healing and angiogenesis. Since platelets are among the first cells involved in haemostasis following tissue injury, our data showing that ligation of CD40 by CD154 induces a procoagulant phenotype on vascular endothelial cells suggests that platelets may play an important role in the induction of wound healing. PMID- 9869176 TI - Effect of the antioxidants selenium and beta-carotene on HIV-related endothelium dysfunction. AB - Patients infected with HIV are at increased risk of atherosclerosis, and have evidence of endothelium dysfunction. The hypothesis was tested that HIV-related endothelium dysfunction is related to loss of antioxidants. This was done by the supplementation of the antioxidants selenium and beta-carotene. We supplemented the diet of 10 HIV-seropositive subjects with 100 microg selenium daily, 11 subjects with 30 mg beta-carotene twice daily while 15 subjects were not supplemented. Plasma was obtained at outset and after a year, and tested by ELISA for endothelial cell, platelet and inflammatory markers. The non-supplemented patients experienced increases in von Willebrand factor and soluble thrombomodulin (both p <0.01). There were no changes in any of the indices in the patients taking selenium or beta-carotene. Increased von Willebrand factor and soluble thrombomodulin in the non-supplemented patients imply increased damage to the endothelium over the year of the study. Therefore we interpret the lack of increase in the patients taking antioxidants as evidence of the protection of the endothelium by these agents. PMID- 9869177 TI - Intermediates of prothrombin activation induce intracellular calcium mobilization in rat aortic smooth muscle cells. AB - While effects of alpha-thrombin have been well characterized in different cell types, the biological function of intermediates of prothrombin activation is still undefined. Meizothrombin could be shown to be a potent agonist for vascular contraction which seems to be mediated by an interaction with the vascular smooth muscle. To explore this effect at intracellular signaling level, we used rat aortic smooth muscle cells and investigated the effect of the intermediates formed by cleavage of human prothrombin with ecarin, the prothrombin activator from Echis carinatus venom, on mobilization of free intracellular calcium. The ecarin-activated prothrombin induced very rapidly transient calcium mobilization in SMC's comparable to that observed with alpha-thrombin. We conclude that meizothrombin/meizothrombin desF1 (MT/MT desF1) are the most likely candidates for this effect. Furthermore, our results suggest the involvement of PAR-1-type thrombin receptors in MT/MT desF1-induced calcium signaling in rat aortic smooth muscle cells. PMID- 9869178 TI - Melagatran, a low molecular weight thrombin inhibitor, counteracts endotoxin induced haemodynamic and renal dysfunctions in the pig. AB - Coagulation and fibrinolysis are crucial in septic shock and inhibition of thrombin may be beneficial in this circumstance. Since porcine endotoxaemia has been found to replicate severe septic shock, a low molecular weight thrombin inhibitor, melagatran, was infused during the first 3 out of 6 h of endotoxaemia in pigs. Plasma creatinine (p <0.01) and urinary output (p <0.05) were less affected in the melagtran + endotoxin group (n=6) as compared to endotoxaemic controls (n=9). The left ventricular stroke work index, systemic vascular resistance index and oxygen extraction were all less affected (p <0.05) by endotoxin during the infusion of melagatran. The plasma concentration of melagatran declined with an apparent plasma half-life of 5 h as soon as the infusion was stopped. APTT, however, continued to increase after the infusion of melagatran had stopped and reached a maximum of 113 s at 5 h (baseline 17 s). APTT in endotoxaemic control pigs reached a maximum of 22 s. Thus, melagatran may counteract some consequences of endotoxaemia. PMID- 9869180 TI - Homozygous G20210A prothrombin gene mutation without thromboembolic events: a case report. PMID- 9869179 TI - Homozygous G20210A transition in the prothrombin gene associated with severe venous thrombotic disease: two cases in a French family. PMID- 9869181 TI - The 4G/5G polymorphism of PAI-1 promoter gene and the risk of myocardial infarction: a meta-analysis. PMID- 9869182 TI - Increased soluble P-selectin in peripheral artery disease: a new marker for the progression of atherosclerosis. PMID- 9869183 TI - Steroid hormone-responsive secondary factor X deficiency. PMID- 9869184 TI - Recurrent adenocarcinoma of prostate presenting as acquired haemophilia A. PMID- 9869185 TI - Reduction of acquired high titer factor VIII antibodies by extracorporeal antibody-based immunoadsorption without additional immunosuppressive therapy. PMID- 9869186 TI - Is a change of factor VIII product a risk factor for the development of a factor VIII inhibitor? PMID- 9869187 TI - The Second British Standard for batroxobin (moojeni) PMID- 9869188 TI - Monoclonal antibodies in the treatment of human B-cell malignancies. AB - Monoclonal antibody technology emerged in the 1970's and was greeted by a wave of optimism. Many believed this new form of therapy would be effective in the treatment of human cancers. Early clinical trials in B-cell lymphomas demonstrated both the potential and limitations of unlabeled murine monoclonal antibody therapy, and taught us valuable lessons regarding the importance of the antibody structure, and nature of the targeted antigen. Since that time modifications in antibody structure and careful selection of target antigen have improved the clinical efficacy of these agents. Clinical trials using humanized antibodies have demonstrated that human/mouse chimeric antibodies and humanized antibodies have enhanced anti-tumor activity, decreased immunogenicity, and a very favorable toxicity profile. Radiolabeled monoclonal antibodies can induce durable remissions in lymphoma with toxicity limited largely to bone marrow suppression. Clinical trials with immunotoxins have demonstrated anti-tumor activity but also have been associated with significant toxicity. Standard treatment options for B-cell lymphoma will soon include antibody-based therapies. Further basic and clinical research is needed so we can understand more thoroughly the mechanisms responsible for the observed anti-tumor effects, and explore more extensively the best approach to their clinical use. PMID- 9869189 TI - Human herpesvirus 6 infection and associated pathogenesis following bone marrow transplantation. AB - Human herpesvirus 6 (HHV-6) infections following bone marrow transplantation (BMT) have been shown to be associated with fever, skin rash, graft versus host disease, encephalitis, delay in engraftment, marrow suppression, and pneumonia. Unfortunately several of these studies were case reports and although the results were suggestive they prompted us to study these pathological events systematically. These associations were primarily based on either HHV-6 isolation, HHV-6 DNA detection, antigen detection or increases in HHV-6 specific antibodies. HHV-6 activity was more frequent during the post- rather than the pre transplantation period. All HHV-6 isolates from BMT patients have been shown to be variant B. A better understanding of HHV-6 associated pathogenesis gained by larger prospective trials is needed to facilitate proper treatment of cases of idiopathic illnesses or those associated with symptoms (fever, skin rash) similar to those caused by HHV-6. PMID- 9869190 TI - Prevention and treatment of hepatic venocclusive disease after high-dose cytoreductive therapy. AB - Venocclusive disease of the liver (VOD) is one of the most common and serious complications following stem cell transplantation. High-dose chemotherapy or chemoradiotherapy injures the structures of Zone 3 of the liver acinus and produces the clinical syndrome of hepatomegaly or right upper quadrant pain, jaundice, and fluid retention. VOD occurs in up to 54% of stem cell transplant recipients and is fatal in 25-50% of them. While the clinical signs of VOD usually manifest during the first post-transplant week, late presentation can occur. The purpose of this review is to discuss the manifestations and pathophysiology of VOD and the options for prevention and treatment. PMID- 9869191 TI - Vitamin D analogs, leukemia and WAF1. AB - Vitamin D compounds induce differentiation of human leukemic cells and have potential for the treatment of leukemia. In this review we summarize some of the basic mechanisms underlying the action of vitamin D compounds. A variety of vitamin D analogues were synthesized until now, some of which have enhanced antileukemic activity and a decreased propensity to cause hypercalcemia. Most actions of vitamin D compounds are mediated by nuclear receptors. In vivo, vitamin D binding protein interacts with free vitamin D compounds. Both in normal and leukemic cells, vitamin D compounds cause a differentiation to monocytes and macrophages. A variety of genes are regulated by vitamin D compounds. Recently, the cell cycle inhibitory gene p21/WAF-1/CIP-1 was characterized. The expression de novo of WAF-1 in blasts of acute myelogenous leukemia is an independent factor of unfavorable prognosis. In HL-60 leukemic cells treated with vitamin D analogs, WAF-1 can be induced by nano- or picomolar concentrations of vitamin D analogs and correlates with the induction of a differentiated phenotype. When vitamin D analogs are combined in-vitro with retinoids, an irreversible differentiation is observed. Clinical trials of vitamin D analogs are indicated in the situation of minimal residual disease and in combination with standard chemotherapy. PMID- 9869192 TI - Biology of CD34+CD38- cells in lymphohematopoiesis. AB - Since the discovery of the CD34 stem/progenitor cell antigen, considerable progress has been made in further purifying human lymphohematopoietic stem cells (HSC). These studies have identified a number of antigens which can be targeted to subfractionate the CD34+ cell population. In particular, several lines of evidence suggest that the rare CD38(-)subpopulation of CD34+ cells may be enriched in HSC. This review briefly summarizes relevant knowledge concerning the CD38 molecule and the results of in vitro and in vivo studies of CD34+38(-)cells. Possible clinical uses for purified CD34+38(-)cells are outlined. PMID- 9869193 TI - Involvement of granulocyte colony-stimulating factor in proliferation of adult T cell leukemia cells. AB - Granulocyte-colony stimulating factor (G-CSF) was originally found to induce proliferation and differentiation of normal granulocyte progenitors. Recent studies demonstrated that G-CSF induces growth of some malignant cells, including lymphoid cells. G-CSF is now widely and successfully used to treat neutropenia induced by intensive chemotherapy, and the responsive growth of malignant cells becomes a major clinical issue. Adult T-cell leukemia (ATL) is a malignant lymphoid disease of T cells, etiologically associated with human T cell lymphotropic virus type I (HTLV-I). We demonstrated that primary ATL cells in about 80% of patients expressed cell surface G-CSF receptor (G-CSFR). Our recent data also show that ATL cells from a third of the patients show responsive growth to G-CSF ex vivo. Several patients whose ATL cells proliferated in response to G CSF showed a significant increase of the ATL cell count after administration of G CSF in vivo. These observations suggest caution for it's routine clinical use in ATL. The molecular mechanism of G-CSF responsive growth of ATL cells is obscure, however the population of G-CSFR expressing cells is larger in responsive cases than in nonresponsive cases. Expression of G-CSFR on ATL cells may relate to the expression of Tax protein encoded by HTLV-I. Precise studies on G-CSFR signaling in ATL cells are necessary for the safe use of G-CSF routinely for ATL patients. PMID- 9869194 TI - History and classification of human leukemia-lymphoma cell lines. AB - Continuous human leukemia-lymphoma cell lines have become indispensable tools in hematological research since the establishment of the first human lymphoma cell line Raji in 1963. We summarize here historical landmarks in the establishment of unique leukemia-lymphoma-derived cell lines from the various cell lineages; their special importance in hematopoietic research is emphasized. The first cell lines were derived from African Burkitt lymphomas and were found to integrate the Epstein-Barr virus in their genome leading to the discovery and isolation of this virus. However, it was later recognized that not every cell line derived from a patient with leukemia-lymphoma represents a malignant cell line as residual normal B-lymphocytes can also be immortalized by EBV infection. During the following 20-30 years many other types of hematopoietic cell lines, commonly derived from hematopoietic neoplasms, were established. These panels of cell lines now span almost the whole spectrum of hematopoietic cell lineages (except for dendritric cells) and the various distinct stages of differentiation along the respective cell axes. From early on, cell lines became important tools for basic and clinical hematological research, initially mainly in the field of immunology, but later expanding to other areas also. It became apparent that leukemia-lymphoma cell lines are of monoclonal origin, are arrested at a discrete maturational stage during differentiation in each lineage, and show sustained and growth factor-independent or -dependent unlimited proliferation. Categorization of cell lines might best be based on the physiological stages of hematopoietic differentiation in the various cell lineages. For an adequate classification, detailed characterizations of both the cell lines and the primary cells from which the cell lines originated are absolutely mandatory. In summary, the availability of large numbers of continuous leukemia-lymphoma cell lines has greatly facilitated clinical and immunobiological studies of normal and malignant hematopoiesis. Human leukemia-lymphoma cell lines will continue to provide exquisite model systems for many biomedical disciplines. PMID- 9869195 TI - The use of the polymerase chain reaction to predict for subsequent relapse in unrelated marrow transplantation for chronic myelogenous leukemia. AB - Disease relapse is one of the causes of treatment failure in patients who undergo allogeneic bone marrow transplantation (BMT) for chronic myelogenous leukemia (CML). Emerging data suggest that accepted therapies for patients who relapse such as interferon and donor lymphocyte infusions may be more efficacious and potentially less toxic if given to patients with minimal disease burdens. The effective application of this strategy may therefore be dependent on the early detection of relapse posttransplant using sensitive assays such as the polymerase chain reaction (PCR) for the presence of the bcr/abl transcript. To that end, we have been interested in determining the predictive value of PCR for subsequent relapse in unrelated marrow transplants for CML. In this review, we update our experience in a cohort of 57 patients transplanted with T cell depleted unrelated marrow grafts who were subsequently followed for relapse by PCR. PMID- 9869196 TI - 5q(-)survival: importance of gender and deleted 5q bands and survival analysis based on 324 published cases. AB - Survival of 5q(-)patients is known to depend on patient age, diagnosis, and acquisition of additional chromosome aberrations. The possible importance of the deleted 5q bands is unknown; nor is it known if the same or different factors are of prognostic importance in 5q(-)MDS and 5q(-)AML. In order to obtain material of sufficient size to answer these and other questions the literature was searched for 5q(-)cases with data on survival. Only MDS and AML cases with interstitial deletions were included. The results confirmed the prognostic importance of the factors mentioned above, but produced the following new data: In MDS, but not in AML, loss of the band 5q12 is an independent adverse prognostic factor; female patients survive longer in MDS while in AML men are the longer survivors; del(5)(q13q31) was found to be associated with MDS, 5q(-)as the sole aberration, and a long survival; on the other hand many del(5)(q12q32) cases showed AML, presence of additional aberrations, and short survivals. On the basis of these results the following hypotheses are proposed: 1) the female preponderance in 5q( )MDS is due to longer female survival, and not to increased female incidence, and 2) some 5q deletions have a high diagnostic specificity, affect the rate of development of additional chromosome aberrations and thus in fact are major prognostic factors. PMID- 9869197 TI - Potential of autologous immunologic effector cells for prediction of progression of disease in patients with chronic myelogenous leukemia. AB - In autologous bone marrow transplantation, immunologic effector cells such as lymphokine activated killer (LAK) cells may be useful for purging of bone marrow since these cells might have an additional in vivo effect on tumor cells in contrast to other purging protocols. Recently, immunologic effector cells termed cytokine-induced killer (CIK) cells have been shown to be more useful than LAK cells for purging of autologous BM in the context of autologous BMT. Here, we show that the expression of bcr/abl in CIK cells generated from patients with CML correlates with progression of disease in individual patients. In addition, progression of disease from chronic phase to accelerated phase could be predicted in two patients by studying the expression of bcr/abl in CIK cells generated from CML patients. Thus, it might be possible to use CIK cell generation for the prediction of progression of disease in CML patients. PMID- 9869199 TI - C-reactive protein serum level is a valuable and simple prognostic marker in non Hodgkin's lymphoma. AB - Interleukin-6 plays a central role in normal B-cell maturation and in proliferation of some B-cell malignancies including multiple myeloma and some non Hodgkin's lymphomas (NHL). Furthermore, this cytokine also plays a major role in acute phase response by mediating synthesis of acute phase proteins such as C reactive protein (CRP). In order to evaluate the exact role of CRP serum level as a simple prognostic factor, we analyzed CRP and IL-6 serum levels in 39 patients with NHL. Eleven patients had low grade NHL, 15 intermediate grade NHL, and 13 high grade NHL. Thirty percent of the patients presented detectable IL-6 serum levels (mean+/-SD: 33.6+/-95.2 U/ml, range: 0 to 500). Increased serum CRP levels were found in 42% of the patients with a mean of 29.2+/-41.97 mg/l] (range: 0 to 129). Thirty seven patients were studied for both markers. Three groups of patients were determined. One with low IL-6 and CRP serum levels (N=21), a second with high level of both markers (N=10), and the third with high serum CRP levels alone (N = 5). Only one patient had high level of serum IL-6 with no detectable CRP. The correlation of serum IL-6 and CRP levels with patient survival was investigated. Median survival in the group with low IL-6 level was not reached. 67% of patients of this group were still alive at 32 months from diagnosis. The group of patients with detectable IL-6 had a median of survival of 12 months (p<0.025). The survival of patients with a CRP<10 mg/l was not reached. 75% of patients survive at 32 months from diagnosis, whereas the group with higher CRP level reached a median survival at 8.5 months (p<0.009). As expected, on univariate analysis, there is a significant relationship between CRP and IL-6 levels (p<0.00017), and CRP levels and B symptoms (p<0.001). Furthermore there is a significant relationship between CRP and LDH levels (p<0.042).These results indicated that CRP may be considered as a valuable and easy prognostic biomarker of NHL. PMID- 9869198 TI - A phase II study of interleukin-2 in 49 patients with relapsed or refractory acute leukemia. AB - In this report we present the results of a multicenter phase II study of high dose recombinant Interleukin-2 (rIL-2) in patients with refractory or relapsed acute leukemia. Forty-nine patients with acute myeloid leukemia (AML: 30 patients) or acute lymphoblastic leukemia (ALL: 19 patients) were included. Median age was 30 years (range: 4-71). Four patients were treated for primary refractory disease and 45 for relapsed disease (16 patients > 2nd relapse). Twenty-four patients (49%) had previously received bone marrow transplantation (allogeneic: 5, autologous: 19). Patients were scheduled to receive three 5-day cycles of rIL-2 given every other week. rIL-2 was administered as bolus I.V. infusion of 8 x 10(6) UI/m2 every 8 hours during cycle I and every 12 hours during cycles 2 and 3. Patients received a mean of 76% of rIL-2 planned dose. Main toxicity was hematologic (grade IV thrombopenia: 84%). Hemodynamic and metabolic toxicities lead to treatment discontinuation in 10 patients (20%). Strong immune activation was achieved including a significant increase in activated T-cells and Lymphokine-Activating-Killer cell (LAK) activity. Twenty seven out of 30 AML patients could be evaluated for response: 2(7%) achieved complete remission (CR) which lasted 3 and 4 months. No response was observed in the 18 assessable ALL patients, most of whom (77 %) presented absolute drug resistance. These results show that this high dose rIL-2 regimen induces significant biological effects and provides some anti-leukemic activity in patients with advanced leukemia. Considering the severe toxicity observed and the limited remission rate achieved here, rIL-2 does not appear to be a valuable therapeutic option for such patients. However, the undoubted anti-leukemic activity of this cytokine invites further investigation especially in the minimal residual disease situation. PMID- 9869200 TI - DIZE (dexamethasone, idarubicin, and continuous infusion of ifosfamide and etoposide): an effective and well-tolerated new regimen for patients with relapsed lymphoma. AB - We performed a phase II study of dexamethasone, ifosfamide, idarubicin and etoposide (DIZE) in patients with relapsed or refractory Hodgkin's (HL) and non Hodgkin's lymphoma (NHL). The regimen consisted of dexamethasone (20 mg i.v. days 1-4), idarubicin (8 mg/m2 i.v. days 1+2), continuous infusion (c.i.) of ifosfamide (1,000 mg/m2 days 1-4), and c.i. etoposide (60 mg/m2 days 1-4). G-CSF (5 microg/kg) was used to support neutrophil recovery from day 5. In older patients (> 60 years) the dosage of idarubicin and ifosfamide was reduced to 75% in the initial cycle. Fourty six patients (pts) were treated with a total of 131 cycles. Sixteen pts were primary resistant and 30 were relapsed. Median age was 54.3 years (range 22-75). The median number of different prior chemotherapies was 1.7 (range 1 to 5). 31/46 (67.4%) pts had advanced disease (stage III or IV); 19/46 had B symptoms. Of 43 evaluable pts the response rate was 58.1% including 11 complete remissions (CR) and 14 partial remissions (PR). Mean duration of response was 8 months (1-30+). DIZE was more effective in relapsed than in refractory high-grade NHL (74 % vs 16.6%; p < 0.001). Of four heavily pretreated pts with HL, one obtained CR and two PR (response rate 75%). Myelosuppression was generally moderate with a mean duration of leukocytopenia < 1,000/microl of 2.5 days (range 0-18) and of thrombocytopenia < 25,000/microl 1.5 days (range 0-17). One patient died of uncontrollable infection in treatment related neutropenia. No other serious toxicities apart from alopecia were observed. We conclude that DIZE is safe and effective in heavily pretreated pts with relapsed lymphoma. The continuous infusion of cytostatic drugs such as that used in the new DIZE protocol might reduce hematotoxicity. PMID- 9869201 TI - Incidence and time trends in Hodgkin's disease: from parts of the United Kingdom (1984-1993). AB - Over 3,000 cases of Hodgkin's Disease diagnosed between 1984-93 were used to examine incidence and time trends. These data are part of the Leukaemia Research Fund's specialist Data Collection Study, which is the only large, population based data set of its type in Europe. The age specific incidence curves showed different patterns for nodular sclerosis contrasted with all other subtypes combined (non-nodular sclerosis). For nodular sclerosis, there was a female excess for young adults, while for non-nodular sclerosis a gradual rise in incidence with age in both sexes was observed. Incidence varied over time, showing a complex pattern with a decreasing trend in males in all Rye-subtypes and no significant change among females diagnosed with nodular sclerosis. These complex patterns of change are different from those seen in other countries. It is concluded that the results provide clear evidence of the heterogeneity of Hodgkin's disease between the sexes and between subtypes, which should be taken into account in future studies. PMID- 9869202 TI - The expression of T cell related costimulatory molecules in multiple myeloma. AB - Presentation of tumour antigen by malignant cells not expressing costimulatory molecules is considered to be a major cause of the failure of the host's immune response against tumours. This study has determined the expression of the B7 family of costimulatory molecules on malignant plasma cells and the expression of the counter receptor molecules, CD28 and CD152 (CTLA-4), on T cells of patients with multiple myeloma. CD28 expression was present on most CD4 cells but was lower on CD8 cells especially from those patients who also showed evidence of expanded T cell clones (median 40%. z=2.4; p<0.02). CD152 expression was increased in 50% (9/18) of patients with myeloma. CD80 (B7-1) expression was present on the plasma cells of only 1 of 27 samples but CD86 (B7-2) expression within the normal range was present on the plasma cells of 14 of 27 samples. Primitive plasma cells (CD38++ CD45++) had a higher expression of CD86 (median 78%) than mature plasma cells (CD38++ CD45-) (median 19%, z=3.7; p<0.01). Thus patients with expanded T cell clones have a downregulated T cell CD28 expression and lack B7-1 expression on their malignant plasma cells. These results are consistent with the concept that engagement of the T cell receptor by tumour antigen on B7-1 deficient malignant plasma cells would result in T cell anergy rather than productive immunity. PMID- 9869203 TI - Anti-CD16/CD30 bispecific antibodies as possible treatment for refractory Hodgkin's disease. AB - Fifteen patients with refractory Hodgkin's disease were treated in a phase I/II dose escalation trial with the NK-cell activating bispecific monoclonal antibody HRS-3/A9 which is directed against the Fcgamma-receptor III (CD16 antigen) and the Hodgkin's associated CD30 antigen, respectively. HRS-3/A9 was given four times every 3-4 days starting with 1 mg/m2. The treatment was well tolerated and the maximum tolerated dose was not reached at 64 mg/m2, the highest dose given due to limited amounts of HRS-3/A9 available. Mild to moderate side effects occured in six patients and consisted of fever, pain in involved lymph nodes, and a maculopapulous rash. Median counts of NK-cells and of all lymphocyte subsets were considerably decreased in the patients before therapy and showed no consistent changes under therapy. Eight patients developed human anti-mouse immunoglobulin antibodies, and five patients showed an allergic reaction after attempted retreatment. One complete and one partial remission (lasting 6 and 3 months, respectively), three minor responses (lasting 1 to 15 months), two disease stabilizations (for 2 and 17 months, respectively), and one mixed response were achieved. There was no clearcut dose-side effect or dose-response correlation. Our results encourage further clinical trials with this novel immunotherapeutic approach and emphasize the necessity to reduce the immunogenicity of the murine bispecific antibodies. PMID- 9869204 TI - Reassessment of non-hodgkins's lymphoma with a "nodular" growth variant: a clinicopathologic study of follicular, mantle cell and marginal zone lymphomas prospectively diagnosed with multiparameter analyses. AB - Although three subtypes of non-Hodgkin's lymphoma (NHL), follicular lymphoma (FL), mantle cell lymphoma (MCL) and marginal zone lymphoma (MZL), are now well recognized as independent categories, their biological behavior has not been fully compared. One of the reasons for this may be that subclassification by histological examination alone is often difficult since they all have a common variant of a "nodular" growth pattern and occasionally show similar cytological morphology. Recently, we reviewed patients with FL, MCL and MZL, who were prospectively diagnosed, using multiparameter analyses with unfixed fresh biopsy materials. Of 407 NHL patients, 101 (24.8%) belonged to these three categories and 80 could be followed; FL (n=27), MCL (n=27) and MZL (n=26). Twenty eight cases with diffuse large B-cell (DL-B) lineage lymphoma were selected as control at random. The frequency of the MCL patients with performance status (PS) 2 to 4 (41%) was significantly higher than MZL patients (4%) [P< 0.001]. The 3 year survival rate with FL, MCL, MZL and DL-B was 71.5%, 57.4%, 93.3% and 53.1%, respectively. The survival rate for MZL was significantly better than both FL (p = 0.048) and MCL (p = 0.0085). Significant differences were also found in the overall survival rates among the four risk groups as defined by the International Index [I2](low, low-intermediate, high-intermediate and high; 97.4%, 79.6%, 39.4% and 18.2%, respectively). A multivariate analysis revealed that the International Index may be a significant predictor for short survival (p=0.0001) in the patients with FL, MCL or MZL. These results suggest that MZL shows an apparently better prognosis than FL and MCL and is found to be a prognostically independent category. In contrast, the clinical outcome in MCL is the worst among the three subtypes and was closer to that of DL-B. The International Index can be applied to a wide spectrum of NHL, including MCL, MZL and FL, to and can predict prognosis in these cases. PMID- 9869205 TI - Expression of deoxycytidine kinase (dCK) gene in leukemic cells in childhood: decreased expression of dCK gene in relapsed leukemia. AB - Competitive RT-PCR was used to determine the quantitative variation in the expression of deoxycytidine kinase (dCK) gene in childhood leukemic cells. The degree of dCK gene expression varied over a 50-fold range. In two cases in which both primary and relapsed leukemic cells were analysed, decreased expression of dCK gene was found in relapsed leukemic cells. The sequence variation analysis using bisbenzimide/polyethylene glycol electrophoresis demonstrated no sequence alteration of dCK cDNA in all cases. These results indicate that the expression of dCK gene varies in patients and suggests decreased expression of the dCK gene as one of the mechanisms responsible for clinical resistance to ara-C. PMID- 9869206 TI - HTLV-I infection among relatives of patients with adult T-cell leukemia/lymphoma in Brazil: analysis of infection transmission. AB - We examined the presence of HTLV-I infection among 66 family members of 13 patients with well documented ATL to investigate the routes of HLTV-I transmission in a Southeast region of Brazil. HTLV-I infection was screened by an enzyme immunossay (ELISA) test and all repeatedly positive or indeterminate ELISA samples were further tested by a Western-Blot (WB) technique. Indeterminate and inconclusive WB samples were confirmed by a polymerase chain reaction (PCR). ELISA results showed that 40 (60.6%) individuals were not infected; 16 (24.2%) were positive; and 10 (15.2%) were undetermined. Among 16 ELISA positive subjects, 14 (87.5%) were confirmed to be positive by WB while 2 (12.5%) showed inconclusive results. Based on the laboratory data, questionnaire analysis, and family/epidemiological studies, we concluded that HTLV-I vertical transmission occurred in 6 of the 13 families. In 3 of these 6 families, the horizontal transmission also could be demonstrated. An isolated horizontal transmission was detected in one family, and in 6 families we did not find any infected family member. All HTLV-I-infected persons were clinically asymptomatic. The occurrence of an effective HTLV-I vertical transmission detected by the present study suggest that HTLV-I infection is endemic in the Southeast region of Brazil. Consistent with the modes of transmission, the HTLV-I antibody seroprevalence was greater in relatives of ATL patients than in the general blood donor Brazilian population (0.4%). In addition, the present data suggest that HTLV-I carries a high infectivity rate but a low virulence. PMID- 9869207 TI - Aggressive NK cell lymphoma preceded by a ten year history of neutropenia associated with large granular lymphocyte lymphocytosis. AB - We report a case of aggressive natural killer (NK) cell lymphoma in an 82 year old man who first presented 10 years earlier with neutropenia in association with a large granular lymphocyte (LGL) lymphocytosis. The diagnosis of NK cell lymphoma was made on the basis of morphological and immunological characteristics (CD3-CD56+) found on skin biopsy of one of multiple skin nodules which subsequently developed in association with splenomegaly, thrombocytopenia and continuing neutropenia. In addition there was BM infiltration and a cytogenetic abnormality [add(6)(p25)] was detected. Combination chemotherapy led to an initial clinical response but a relapse occurred shortly afterwards and the patient died 8 months later from infection whilst neutropenic following re introduction of chemotherapy. Previously reported cases of aggressive NK cell lymphoma have shown a young male predominance with a rapidly progressive clinical course and without evidence of a preceding chronic phase of LGL lymphocytosis and neutropenia. PMID- 9869208 TI - Primary cardiac lymphoma in an immunocompetent woman. AB - We report a fatal primary cardiac non-Hodgkin's lymphoma in a 62 years old immunocompetent woman presenting with tamponade and complete atrioventricular block. CT-scan, echocardiography and autopsy examination showed a tumor largely infiltrating the heart without extracardiac involvement. A surgical biopsy revealed high grade B-cell non-Hodgkin's lymphoma with a misleading myelomonocytic CD68 (KPI) expression. Polymerase Chain Reaction analysis revealed a clonal rearrangement of the immunoglobulin heavy chain gene and confirmed the B cell origin of the lymphoma. Our report also emphasizes the role of immunohistochemical and molecular techniques in the diagnosis. PMID- 9869210 TI - Teaching cases from the Royal Marsden and St Mary's Hospitals. Case 18: Severe anaemia and thrombocytopenia with red cell fragmentation. PMID- 9869209 TI - Myelodysplastic syndrome with myelofibrosis and basophilia: detection of trisomy 8 in basophils by fluorescence in-situ hybridization. AB - We report a case of myelodysplastic syndrome (MDS) with myelofibrosis and pulmonary tuberculosis who had marked basophilia in the peripheral blood. A clonal karyotypic abnormality characterized by trisomy 8 was demonstrated by cytogenetic analysis. By correlation of cell morphology with results of fluorescence in situ hybridization using a chromosome 8 probe, we demonstrated that the basophils were not reactive but belonged to the neoplastic MDS clone. PMID- 9869211 TI - Mini-ICE protocol is better than high-dose hydroxyurea to mobilize Ph-negative cells in earlier phases of chronic myelogenous leukemia. PMID- 9869212 TI - Randomized clinical trial on the combination of preoperative irradiation and surgery in the treatment of adenocarcinoma of gastric cardia (AGC)--report on 370 patients. AB - PURPOSE: An attempt was made to define the role of radiotherapy before operation for AGC. METHODS AND MATERIALS: From January 1978 to May 1989, a prospective randomized trial on preoperative radiotherapy (R+S) vs. surgery alone (S) for AGC was carried out in 370 patients. Patients were randomized into a combined group (R+S, 171 patients) or a surgery alone group (S, 199 patients) by the envelope method. 8-MV photon or telecobalt was used for the preoperative radiation therapy, using anterior-posterior opposing parallel fields to deliver 40 Gy to the cardia, lower segment of the esophagus, fundus, lesser curvature, and hepatogastric ligament. Surgery was performed after 2 to 4 weeks rest. RESULTS: The 5- and 10-year survival rates of the R+S Group and the S Alone Group were 30.10% and 19.75%, 20.26% and 13.30%, respectively. The survival curves of these two groups diverged right from the beginning after the operation over the ninth year. Statistics by Kaplan-Meier log rank test proves that the difference is significant (chi2 = 6.74, p = 0.0094). The immediate results were: resection rate 89.5% and 79.4% (p < 0.01); pathologic stage after resection T2 12.9% and 4.5% (p < 0.01), T4 40.3% and 51.3% (p < 0.05), lymph node metastasis rates 64.3% and 84.9% (p < 0.001); operative mortality rates 0.6% and 2.5%; intrathoracic leak rates 1.8% and 4.0%, respectively. The causes of failure were: local uncontrol and recurrence 38.6% vs. 51.7% (p < 0.025), regional lymph node metastasis 38.6% vs. 54.6% (p < 0.005), distant metastasis 24.3% vs. 24.7%. CONCLUSION: Preoperative radiation therapy is able to improve the results of surgery for adenocarcinoma of the gastric cardia. PMID- 9869213 TI - Prognostic implications of downstaging following preoperative radiation therapy for operable T3-T4 rectal cancer. AB - PURPOSE: To evaluate the prognostic value of tumor downstaging after preoperative radiation for resectable rectal cancer. METHODS AND MATERIALS: Eighty-eight patients with non-metastatic resectable rectal cancers (76 T3 and 12 T4) were treated with preoperative irradiation. Median dose was 40 Gy (30-46 Gy) delivered over 32 days (range 11-40). Seventeen patients received preoperative chemotherapy, two courses of 5-fluorouracil (5FU) 350 mg/m2/day and folinic acid 20 mg/m2/day; 5 days per week during the first and fifth weeks of radiotherapy. Surgery was performed with a mean delay of 46 days after completion of irradiation and included 66 abdominoperineal resections and 22 anal sphincter preserving procedures. Postoperative chemotherapy was administered in 44 patients. RESULTS: Histological tumor stages were: complete histological response in 7%, pT2N0 in 19%, pT3N0 in 46%, and pT2-3N1 in 28%. Tumor downstaging occurred in 26%. No predictive factor of downstaging was statistically significant. The median follow-up was 33 months. The 3- and 5-year cancer-specific survival rates were 100% for the pT0N0 and pT2N0, respectively, 89% and 68% for pT3N0, and 64% and 0% for pT2T3N1. After preoperative irradiation, the pathological tumor stages remained a prognostic factor. Patients with downstaging (pT0T2N0) had significantly higher cancer-specific survival rates than the group without downstaging: 100% and 80% at 3 years, and 100% and 45% at 5 years; respectively (p = 0.011). The 3- and 5-year recurrence free-survival rates were 94% for the group with downstaging and 56% and 50%, respectively, for the group without downstaging (p = 0.002). CONCLUSION: Downstaging after preoperative irradiation in this retrospective study results in an improvement in local control and survival. PMID- 9869214 TI - Rapid growth of microscopic rectal cancer as a determinant of response to preoperative radiation therapy. AB - PURPOSE: To quantify the dose-time fractionation factors in preoperative radiation therapy for microscopic pelvic deposits of rectal cancer. This provides a biologic basis for understanding and improving the results of adjuvant therapies for this disease. METHODS: The reduction in incidence of pelvic relapses as a function of radiation dose and overall treatment time was determined from the literature. The displacement of dose-response curves to higher doses reflects the growth during radiation treatment of subclinical pelvic deposits which are beyond the future surgical margins. RESULTS: Dose-response curves are steep if the effect of overall duration of radiation therapy is accounted for. The time-related displacement of these steep dose-response curves is consistent with a median doubling time for malignant clonogenic cells of about 4 or 5 days, much faster than the growth rate of the average primary tumor at diagnosis. This rapid growth is evident within the first few days of irradiation, implying that the natural growth rate of these microscopic deposits if fast, and/or that an acceleration of growth follows initiation of radiation injury with a very short lag time. CONCLUSION: Subclinical pelvic deposits of rectal cancer grow rapidly during preoperative radiation therapy with an adverse influence on the rate of pelvic tumor control from protracting the duration of adjuvant treatment. Low doses only offer clinically relevant reduction in risk of pelvic relapses if the overall radiation treatment time is short. For a given overall treatment duration there is a relatively steep dose-response curve, predicting that significant improvements in tumor control are possible. PMID- 9869215 TI - The results of radiotherapy for ependymomas: the Mayo Clinic experience. AB - PURPOSE: This analysis was performed to examine the outcome of patients with histologically confirmed ependymomas of the brain or spinal cord who received postoperative radiotherapy. METHODS AND MATERIALS: Eighty patients with histologically confirmed ependymomas were evaluated retrospectively. All were treated with various combinations of surgery, radiotherapy (RT), and chemotherapy. Follow-up ranged from 5 to 30 years (median 10.4 years). RESULTS: The 5- and 10-year survival rates for the entire study group were 79% and 73%, respectively. Patients with low-grade (1 and 2 of 4) tumors had a 5-year survival rate of 87% as compared to 27% for those with high-grade (3 and 4 of 4) tumors (p < 0.0001). Patients with tumors of the spine had a 5-year survival rate of 97% as compared to 68% for those with infratentorial tumors, and 62% for those with supratentorial tumors (p = 0.03). Patients with myxopapillary ependymomas of the spine had a 5-year survival rate of 100% as compared with 76% for patients with other histological subtypes of ependymoma (p = 0.02). Multivariate analysis revealed that the survival rate was independently associated with tumor grade (p = 0.0007) and histological subtype (p = 0.02). Twenty-eight patients (35%) experienced local failure and 10 patients (13%) developed leptomeningeal seeding. The 5-year leptomeningeal failure rate was 10% in patients with low-grade tumors as compared to 41% for patients with high grade tumors (p = 0.01). CONCLUSION: Patients with low-grade tumors, especially those with myxopapillary subtypes, have high 5-year survival rates when treated with post-operative radiotherapy. High grade ependymomas are associated with a much poorer outcome. New forms of therapy are required to improve the outcome of patients with high-grade ependymomas. PMID- 9869216 TI - Survival of very young children with medulloblastoma (primitive neuroectodermal tumor of the posterior fossa) treated with craniospinal irradiation. AB - PURPOSE: Very young children with medulloblastoma are considered to have a worse prognosis than older children. As radiotherapy remains an important part of the treatment, the adverse prognosis could be due to inadequate radiation treatment rather than biological factors. We analyzed the published literature to examine the impact of radiotherapy on survival in this group. METHODS AND MATERIALS: A Medline search was performed and we reviewed studies of treatment of medulloblastoma where radiotherapy was delivered using megavoltage equipment and the minimum follow-up allowed the calculation of 5-year survival rates. RESULTS: Thirty-nine studies were published between 1979 and 1996 with a treatment including craniospinal irradiation and boost to the posterior fossa. Eleven studies comprising 1366 patients analyzed survival by age at diagnosis. Eight of 11 studies showed a worse 5-year survival for the younger patient group which reached statistical significance in two. There is also a suggestion of a higher proportion of children with metastatic disease at presentation in the very young age group. The usual policy in younger children was to give a lower dose of radiotherapy to the craniospinal axis (CSA) and posterior fossa (PF) with reduction of dose in the range of 15 to 25% compared to standard treatment. As dose reduction to the posterior fossa is associated with worse survival and local recurrence is the predominant site of failure, the major determinant of worse survival in very young children with medulloblastoma may be suboptimal radiotherapy. Protocols including postoperative chemotherapy with delayed, omitted, or only local tumor irradiation do not reach survival rates of protocols with standard radiotherapy, also suggesting a continued importance for irradiation. CONCLUSION: Very young children with medulloblastoma have a worse prognosis than older children. Inadequate radiation dose and technique to the primary tumor region may be a major contributing factor. Current chemotherapeutic regimes alone are not sufficient to compensate for reduced radiation doses and volumes. PMID- 9869217 TI - Phase I and pharmacokinetic study of preirradiation chemotherapy with BCNU, cisplatin, etoposide, and accelerated radiation therapy in patients with high grade glioma. AB - PURPOSE: We conducted a Phase I study of bischloroethylnitrosourea (BCNU), cisplatin, and oral etoposide administered prior to and during accelerated hyperfractionated radiation therapy in newly diagnosed high-grade glioma. Pharmacokinetic studies of oral etoposide were also done. METHODS AND MATERIALS: Patients started chemotherapy after surgery but prior to definitive radiation therapy (160 cGy twice daily x 15 days; 4800 cGy total). Initial chemotherapy consisted of BCNU 40 mg/m2 days 1-3, cisplatin 30 mg/m2 days 1-3 and 29-31, and etoposide 50 mg orally days 1-14 and 29-42, repeated in 8 weeks concurrent with radiation therapy. BCNU 200 mg/m2 every 8 weeks x 4 cycles was given after radiation therapy. RESULTS: Sixteen patients, 5 with grade 3 anaplastic astrocytoma and 11 with glioblastoma were studied. Grade 3-4 leukopenia (38%) and thrombocytopenia (31%) were dose-limiting. Other toxicities were anorexia (81%), nausea (94%), emesis (56%), alopecia (88%), and ototoxicity (38%). The maximum tolerated dose was BCNU 40 mg/m2 days 1-3, cisplatin 20 mg/m2 days 1-3 and 29-31, and oral etoposide 50 mg days 1-21 and 29-49 prior to radiation therapy and repeated in 8 weeks with the start of radiation therapy followed by BCNU 200 mg/m2 every 8 weeks for 4 cycles. Median time to progression and survival were 13 and 14 months respectively. Responses occurred in 2 of 9 (22%) patients with evaluable disease. In pharmacokinetic studies, all patients achieved plasma concentrations of >0.1 microg/ml etoposide (the in vitro radiosensitizing threshold), following a 50 mg oral dose. The mean +/- SD 2 hr and 6 hr plasma concentrations were 0.92 +/- 0.43 microg/ml and 0.36 +/- 0.12 microg/ml, respectively. Estimated duration of exposure to >0.1 microg/ml etoposide was 10 17 hr. CONCLUSIONS: Preirradiation chemotherapy with BCNU, cisplatin, and oral etoposide with accelerated hyperfractionated radiation therapy in high-grade gliomas is feasible and merits further investigation. Sustained radiosensitizing concentrations can be achieved with low oral doses of etoposide. PMID- 9869218 TI - Efficacy of radiotherapy for malignant gliomas in elderly patients. AB - PURPOSE: Age above 65 years is a strong negative prognostic factor for survival in patients with malignant gliomas (MG) treated with radiotherapy (RT) and its value has been questioned. We analyzed the effect of RT on the survival of elderly patients with malignant gliomas. METHODS AND MATERIALS: We examined 85 consecutive elderly patients with a histological diagnosis of MG. Age ranged from 65 to 81 years (median 70 years). Glioblastoma multiforme (GBM) was diagnosed in 64 patients (75.3%). Surgical treatment included needle biopsy in 32 patients (37.6%). Median postoperative Karnofsky Performance Status (KPS) was 60 (range: 30-100). Survival probability was estimated using Kaplan-Meier method and compared with the log-rank test. Crude and adjusted hazard ratios (HR) were calculated using Cox's regression models. RESULTS: Median survival time for all patients was 18.1 weeks. In multivariate analysis, RT was the only independent prognostic variable for survival (HR: 9.1 [95% CI: 4.5-18.7]). Forty-two patients did not start RT mostly due to low KPS (<50). The median survival of the 43 patients who started RT was 45 weeks. In these patients, Cox multivariate analysis indicated that age was independently associated with prolonged survival (HR: 2.85 [95% CI 1.31-6.19]). Median survival of patients age 70 years and younger was 55 weeks compared with 34 weeks for patients older than 70 years. CONCLUSIONS: The overall survival for elderly patients with MG is poor. RT seems to improve survival in patients up to 70 years, but in older patients treated with RT the survival is significantly shorter. PMID- 9869219 TI - Outcome in elderly patients undergoing definitive surgery and radiation therapy for supratentorial glioblastoma multiforme at a tertiary care institution. AB - PURPOSE: To determine the efficacy of definitive surgery and radiation in patients aged 70 years and older with supratentorial glioblastoma multiforme. METHODS AND MATERIALS: We selected elderly patients (> or = 70 years) who had primary treatment for glioblastoma multiforme at our tertiary care institution from 1977 through 1996. The study group (n = 102) included 58 patients treated with definitive radiation, 19 treated with palliative radiation, and 25 who received no radiation. To compare our results with published findings, we grouped our patients according to the applicable prognostic categories developed by the Radiation Therapy Oncology Group (RTOG): RTOG group IV (n = 6), V (n = 70), and VI (n = 26). Patients were retrospectively assigned to prognostic group IV, V, or VI based on age, performance status, extent of surgery, mental status, neurologic function, and radiation dose. Treatment included surgical resection and radiation (n = 49), biopsy alone (n = 25), and biopsy followed by radiation (n = 28). Patients were also stratified according to whether they were optimally treated (gross total or subtotal resection with postoperative definitive radiation) or suboptimally treated (biopsy, biopsy + radiation, surgery alone, or surgery + palliative radiation). Patients were considered to have a favorable prognosis (n = 39) if they were optimally treated and had a Karnofsky Performance Status (KPS) score of at least 70. RESULTS: The median survival for patients according to RTOG groups IV, V, and VI was 9.2, 6.6, and 3.1 months, respectively (log-rank, p < 0.0004). The median overall survival was 5.3 months. The definitive radiation group (n = 58) had a median survival of 7.3 months compared to 4.5 months in the palliative radiation group (n = 19) and 1.2 months in the biopsy-alone group (p < 0.0001). Optimally treated patients had a median survival of 7.4 months compared to 2.4 months in those suboptimally treated (p < 0.0001). The favorable prognosis group had an 8.4-month median survival compared to 2.4 months in the unfavorable group (p < 0.0001). On multivariate analysis, the KPS, RTOG group, favorable/unfavorable prognosis, and optimal treatment/suboptimal treatment were significant predictors of survival. CONCLUSION: Elderly patients with good performance status (> or = 70 KPS) when treated aggressively with maximal resection and definitive radiation had longer survival than those treated with palliative radiation and biopsy. Aggressive treatment in such patients should be considered. PMID- 9869220 TI - Fractionated stereotactic radiotherapy of small intracranial malignancies. AB - PURPOSE: To retrospectively evaluate the effectiveness of fractionated stereotactic radiotherapy (FSRT) in patients with small intracranial malignancies. METHODS AND MATERIALS: From July 1991 to March 1997, 80 patients with a total of 121 brain or skull-base tumors were treated with FSRT alone, and were followed for periods ranging from 3 to 62 months (median 9.8). The majority of patients received 42 Gy in 7 fractions over 2.3 weeks, but in July 1993, protocols using smaller fraction doses were introduced for patients whose radiation-field diameters were larger than 3 cm or whose tumors were close to critical normal tissues. RESULTS: For 64 patients with metastatic brain tumors the overall median survival was 8.3 months and 1-year actuarial survival rate was 33%. Significant prognostic factors were: the presence of extracranial tumors, pre-treatment performance status, and the lung as a primary site. Patients without extracranial tumors prior to FSRT had a median survival of 21.2 months. For seven patients with high-grade glioma, 1-year actuarial local control rate was 75%, with a median survival of 10.3 months. For patients with skull-base tumors the local control was achieved in 6 of 6 patients (100%), with a median survival of 30.7 months. No one suffered from acute complications, but three patients, two of whom had undergone FSRT as the third course of radiotherapy, developed late radiation injuries. CONCLUSION: Overall high local control and low morbidity rates suggest that FSRT is an effective and safe modality, even for those with a history of prior irradiation. However, patients with risk factors should be treated with smaller fraction doses. PMID- 9869221 TI - Assessment of neuropsychological changes in patients with arteriovenous malformation (AVM) after radiosurgery. AB - PURPOSE: The purpose of this study was to investigate neuropsychological effects of radiosurgery in patients with cerebral arteriovenous malformation (AVM), with special focus on attention and memory. This report describes the study setup and presents the first results during a follow-up of up to 1 year. MATERIALS AND METHODS: Seventy-nine patients were studied before, acutely after radiosurgery, and during the regular follow-up (subacute phase: Weeks 6-12, chronic phase: Months 6-12). Radiosurgery was performed using a modified linear accelerator (minimum doses to the target volume: 15-22 Gy, median 20 Gy). Estimated whole brain dose was 0.5 to 2 Gy. Neuropsychological testing included assessment of general intelligence (Wechsler Adult Intelligence Scale), attention (modified Trail-Making Test A, Digit Symbol Test, D2 Test, Wiener Determination Machine) and memory (Rey Auditory Verbal Learning Test, Benton Visual Retention Test). During follow-up, alternate test versions were used. Neuropsychological deficits were defined as a test score of at least one standard deviation (SD) below the mean of the normal distribution. RESULTS: The pretherapeutic evaluation revealed marked deviations from the normal population; 24% had deficits in intelligence (range 23-31% in different subtests), attention (35%, 23-59%) and memory (48%, 31 61%). The overall percentage of aberrant results was reduced by 12% (memory) to 14% (attention) in the chronic phase up to 12 months after therapy. The improvement in test scores was significant (p < 0.05) in 3 of 4 subtests of attention functions. CONCLUSIONS: The acute tolerance of radiosurgery seems to be very good in these patients, showing no relevant increase in number of patients with neuropsychological deficits. Although the long-term follow-up needs to be further increased, our data indicate a tendency to slight improvement in the overall neuropsychological performance of AVM patients in the chronic phase after radiosurgery. PMID- 9869222 TI - Radiotherapy in Ewing's sarcoma and PNET of the chest wall: results of the trials CESS 81, CESS 86 and EICESS 92. AB - PURPOSE: Treatment results and the pattern of relapse were evaluated in the multimodal treatment of Ewing's sarcomas of the chest wall. METHODS AND MATERIALS: In a retrospective analysis, 114 patients with non-metastatic Ewing's sarcoma of the chest wall were evaluated. They were treated in the CESS 81, CESS 86, or EICESS 92 studies between January 1981 and December 1993. The treatment consisted of polychemotherapy (VACA, VAIA, or EVAIA) and local therapy, either surgery alone (14 patients), radiotherapy alone (28 patients) or a combination of both (71 patients). The median follow-up was 46.6 months (range 5-170). A relapse analysis for all patients with local or combined relapses was performed. RESULTS: Overall survival was 60% after 5 years, event-free survival was 50%. Thirty-seven patients had a systemic relapse (32.4%), 11 patients had a local relapse alone (9.6%), and 3 patients had a combined local and systemic relapse (2.6%). The risk to relapse locally after 5 years was 0% after surgery alone, 19% after radiation alone, and 19% after postoperative irradiation. None of the 8 patients with preoperative irradiation have failed locally so far. With the introduction of central radiotherapy planning in CESS 86, local control of irradiated patients improved. Ten of 14 patients with local failure could be evaluated in the relapse analysis: 3 patients had an in-field relapse, 4 patients had a marginal relapse, 2 patients had a relapse outside the radiation fields, and 1 patient failed with pleural dissemination. Six treatment deviations were observed. CONCLUSION: Local control was best after surgery alone in a positively selected group of patients. Local control after radiation or combined radiation and surgery was good. With diligent performance of radiotherapy, it will be possible to further improve the results in the radiotherapy group. PMID- 9869223 TI - Radiation therapy in the management of desmoid tumors. AB - PURPOSE: To evaluate the outcome of patients with extra-mesenteric desmoid tumors treated with radiation therapy, with or without surgery. METHODS AND MATERIALS: The outcome for 75 patients receiving radiation for desmoid tumor with or without complete gross resection between 1965 and 1994 was retrospectively reviewed utilizing univariate and multivariate statistical methods. RESULTS: With a median follow-up of 7.5 years, the overall freedom from relapse was 78% and 75% at 5 and 10 years, respectively. Of the total, 23 patients received radiation for gross disease because it was not resectable. Of these 23 patients, 7 sustained local recurrence, yielding a 31% actuarial relapse rate at 5 years. Radiation dose was the only significant determinant of disease control in this group. A dose of 50 Gy was associated with a 60% relapse rate, whereas higher doses yielded a 23% relapse rate (p < 0.05). The other 52 patients received radiation in conjunction with gross total resection of tumor. The 5- and 10-year relapse rates were 18% and 23%, respectively. No factor correlated significantly with disease outcome. There was no evidence that radiation doses exceeding 50 Gy improved outcome. Positive resection margins were not significantly deleterious in this group of irradiated patients. For all 75 patients, there was no evidence that radiation margins exceeding 5 cm beyond the tumor or surgical field improved local-regional control. Ultimately, 72 of the 75 patients were rendered disease-free, but 3 required extensive surgery (amputation, hemipelvectomy) to achieve this status. Significant radiation complications were seen in 13 patients. Radiation dose correlated with the incidence of complications. Doses of 56 Gy or less produced a 5% 15-year complication rate, compared to a 30% incidence with higher doses (p < 0.05). CONCLUSIONS: Radiation is an effective modality for desmoid tumors, either alone or as an adjuvant to resection. For patients with negative resection margins, postoperative radiation is not recommended. Patients with positive margins should almost always receive 50 Gy of postoperative radiation. Unresectable tumors should be irradiated to a dose of approximately 56 Gy, with a 75% expectation of local control. PMID- 9869224 TI - Cervical carcinoma metastatic to para-aortic nodes: extended field radiation therapy with concomitant 5-fluorouracil and cisplatin chemotherapy: a Gynecologic Oncology Group study. AB - PURPOSE: A multicenter trial of chemoradiation therapy to evaluate the feasibility of extended field radiation therapy (ERT) with 5-fluorouracil (5-FU) and cisplatin, and to determine the progression-free interval (PFI), overall survival (OS), and recurrence sites in patients with biopsy-confirmed para-aortic node metastases (PAN) from cervical carcinoma. METHODS AND MATERIALS: Ninety-five patients with cervical carcinoma and PAN metastases were entered and 86 were evaluable: Stage I--14, Stage II--40, Stage III--27, Stage IVA--5. Seventy-nine percent of the patients were followed for 5 or more years or died. ERT doses were 4500 cGy (PAN), 3960 cGy to the pelvis (Stages IB/IIB), and 4860 cGy to the pelvis (Stages IIIB/IVA). Point A intracavitary (IC) doses were 4000 cGy (Stages IB/IIB), and 3000 cGy (Stages IIIB/IVA). Point B doses were raised to 6000 cGy (ERT + IC) with parametrial boost. Concomitant chemotherapy consisted of 5-FU 1000 mg/m2/day for 96 hours and cisplatin 50 mg/m2 in weeks 1 and 5. RESULTS: Eighty-five of 86 patients completed radiation therapy and 90% of patients completed both courses of chemotherapy. Gynecologic Oncology Group (GOG) grade 3 4 acute toxicity were gastrointestinal (18.6%) and hematologic (15.1%). Late morbidity actuarial risk of 14% at 4 years primarily involved the rectum. Initial sites of recurrence were pelvis alone, 20.9%; distant metastases only, 31.4%; and pelvic plus distant metastases, 10.5%. The 3-year OS and PFI rate were 39% and 34%, respectively, for the entire group. OS was Stage I--50%, Stage II--39%, and Stage III/IVA--38%. CONCLUSIONS: Extended field radiation therapy with 5-FU and cisplatin chemotherapy was feasible in a multicenter clinical trial. PFI of 33% at 3 years suggests that a proportion of patients achieve control of advanced pelvic disease and that not all patients with PAN metastases have systemic disease. This points to the importance of assessment and treatment of PAN metastases. PMID- 9869225 TI - Perioperative morbidity and mortality of high-dose-rate gynecologic brachytherapy. AB - PURPOSE: To determine the 30-day morbidity and mortality rates for patients with an intact uterus undergoing high-dose-rate (HDR) brachytherapy, and to assess risk factors which may predict for these potentially life-threatening complications. METHODS AND MATERIALS: From August 1989 to December 1994, 128 cervical and 41 medically inoperable endometrial cancer patients were treated with 5 outpatient weekly HDR brachytherapy insertions. Patients with cervical cancer also were treated with external beam radiotherapy. Acute events that resulted in either hospitalization (morbidity) or death (mortality) within 30 days of the implant were analyzed. Univariate and multivariate analyses were performed to identify risk factors. RESULTS: Overall there were 16 acute events in 169 patients (9.5%). The overall morbidity and mortality rates for the cervical and endometrial patients were 5.5%, 1.6%, 7.3%, and 9.8%, respectively. The following factors were significant by univariate analysis: age per decade, American Society of Anesthesiologists (ASA) score, Karnofsky Performance Status (KPS), significant medical history, diagnosis of cervical vs. endometrial cancer, and mean time exceeding 160 minutes for the procedure. Since age was the most significant predictive factor (p = 0.0003), bivariate analyses were performed by adjusting for age. In these analyses only ASA and KPS maintained significance, while a positive medical history was of borderline significance (p = 0.07). CONCLUSION: The morbidity and mortality rates observed in gynecologic patients selected for HDR brachytherapy were similar to low-dose-rate, but higher than other HDR reports. Reasons for this include a higher risk population, especially those with medically inoperable endometrial cancer. In the cervical cancer patients, some of the complications may have also been a result of the external beam portion of the radiation. In order to minimize the acute complications observed in the present HDR brachytherapy system, the following changes have been implemented: appropriate patient selection, anesthesiology involvement to monitor conscious sedation for high-risk patients, external beam radiotherapy alone in patients at extremely high risk, deep vein thrombosis (DVT) prophylaxis, use of intraoperative ultrasound, shorter duration in the brachytherapy suite, and preradiation treatment plans (plans executed prior to the insertion) if applicable. Finally, this analysis suggests that these procedures should be performed in a hospital-based setting where appropriate support is available. PMID- 9869226 TI - High-dose-rate brachytherapy alone post-hysterectomy for endometrial cancer. AB - PURPOSE: To evaluate the outcome of post-hysterectomy adjuvant vaginal high-dose rate (HDR) brachytherapy. METHODS AND MATERIALS: A retrospective analysis was performed on a series of 143 patients with endometrial cancer treated with HDR brachytherapy alone post-hysterectomy from 1985 to June 1993. Of these patients, 141 received 34 Gy in four fractions prescribed to the vaginal mucosa in a 2-week period. The median follow-up was 6.9 years. Patients were analyzed for treatment parameters, survival, local recurrence, distant relapse, and toxicity. RESULTS: Five-year relapse free survival and overall survival was 100% and 88% for Stage 1A, 98% and 94% for Stage IB, 100% and 86% for Stage IC, and 92% and 92% for Stage IIA. The overall vaginal recurrence rate was 1.4%. The overall late toxicity rate was low, and no RTOG grade 3, 4, or 5 complications were recorded. CONCLUSION: These results are similar to reported international series that have used either low-dose-rate or HDR brachytherapy. The biological effective dose was low for both acute and late responding tissues compared with some of the HDR brachytherapy series, and supports using this lower dose and possibly decreasing late side-effects with no apparent increased risk of vaginal recurrence. PMID- 9869227 TI - Pulsed dose rate (PDR) brachytherapy as salvage treatment of locally advanced or recurrent gynecologic cancer. AB - PURPOSE: Pulsed dose rate (PDR) brachytherapy is a new treatment option permitting dose distribution optimization in interstitial implants. It possesses the advantage of equipment simplification and radiation protection to the staff, compared to the manually afterloading technique. This study presents the first clinical results from The Finsen Center with PDR-brachytherapy in patients with locally advanced or recurrent gynecologic cancer. METHODS AND MATERIALS: Between June 1993 and August 1996, 34 patients with gynecologic malignancies (22 pelvic recurrences, 12 primary locally advanced) have been treated with external irradiation, four-field box technique, to 46 Gy/23 fractions, 5 F/week and 192Ir interstitial PDR-brachytherapy in pulses of 0.6 Gy, one pulse per hour to a total of 30 Gy. The Martinez Universal Perineal Interstitial Template applicator was used for all implantations. RESULTS: The overall complete response rate was 74%. At median 14 months follow-up (range 3-40) 15 patients were alive with no evidence of disease. Seven of 14 patients with a second recurrence or progressive disease were still alive. The overall 1- and 2-year survival was 71% and 63%, respectively. There was no difference in survival probability when stratifying the patients by primary diagnosis (recurrent vs. primary advanced), relapse locations (central vs. central + pelvic wall mass) or treatment volume. Seventeen chronic grade III complications were observed in 10 patients. Large treatment volumes significantly correlated to severe gastrointestinal complications. Fifteen of 17 chronic grade III complications were observed in patients treated for recurrent disease. CONCLUSION: PDR-brachytherapy in combination with external irradiation is an effective treatment option for patients with locally advanced or recurrent gynecologic cancer, although substantial toxicity is observed in patients with large treatment volumes and recurrent disease. PMID- 9869228 TI - The use of laminarias for osmotic dilation of the cervix in gynecological brachytherapy applications. AB - PURPOSE: Osmotic dilators (laminarias) have been used for gradual nontraumatic dilation of the cervical canal for various intrauterine procedures; however, this technique has not been well accepted in gynecological brachytherapy. The purpose of this study was to evaluate the efficacy of osmotic cervical dilation for brachytherapy in gynecologic cancer patients, without the use of general/regional anesthesia, and to assess patient tolerance, complications, and outcome. METHODS AND MATERIALS: Thirteen brachytherapy procedures were performed in 6 patients with clinical Stages I and II endometrial (5) and Stage IB cervical cancer (1), who were unable to tolerate general/regional anesthesia because of severe medical problems. An osmotic dilator (synthetic laminaria) was inserted into the cervical os 10-12 h before each brachytherapy procedure and removed just before the procedure. Standard Fletcher-Suit-Delclos tandem insertions with vaginal colpostats or cylinders were then performed. Degree of cervical dilation, patient discomfort, procedure time, intra- and postoperative complications were recorded, and local control and survival were assessed. Median follow-up was 31 months (range: 8-35 months). RESULTS: The diameter of the dilated cervical os after laminaria removal was adequate (> or = 5 mm) for tandem insertion, and no additional mechanical dilation was required in all but one procedure (1 of 13). All procedures were performed without general/regional anesthesia. The mean duration of the procedures was 44 min (range, 20-60 min). Discomfort was minimal in all cases. There were no intra- or postoperative complications. All patients maintained local control until death (1 of metastatic disease, 2 of intercurrent disease) or last follow-up (2 with no evidence of disease, 1 alive with metastatic disease). CONCLUSION: This preliminary study suggests that osmotic cervical dilation with a synthetic laminaria is a useful technique to facilitate intrauterine tandem insertion in patients who cannot tolerate general/regional anesthesia. This technique may reduce treatment-associated morbidity, shorten procedure time, and allow the delivery of adequate radiation therapy in this uncommon but challenging patient population. PMID- 9869230 TI - Reduction of radioactive seed embolization to the lung following prostate brachytherapy. AB - PURPOSE: Ultrasound-guided interstitial implantation of radioactive seeds is a common treatment for early stage prostate cancer. One of the risks associated with this therapy is seed embolization to the lung. This paper reports on the incidence and possible adverse effects of seed migration. METHODS AND MATERIALS: Two hundred ninety consecutive patients were treated with permanent radioactive seed brachytherapy for prostate cancer between January 1 and December 31, 1995. One hundred fifty-four patients were treated with iodine-125 (I-125), and 136 patients were treated with palladium-103 (Pd-103). All but one patient had a routine post implant chest radiograph (CXR), leaving 289 evaluable patients. RESULTS: Twenty radioactive seed pulmonary emboli were identified in 17 patients; 3 patients had two emboli each. The radioactive seed pulmonary embolism rate for the entire group of patients was 5.9%. Acute pulmonary symptoms were not reported by any patient in this series. One hundred forty-six study patients were implanted with free seeds alone (136 Pd-103 and 11 I-125), and 143 were implanted with linked seed embedded in a vicryl suture for the peripheral portions of their implants. The radioactive seed embolization rate by patient was 11% (16/146) versus 0.7% (1/143) for free seed implants and implants utilizing linked seeds, respectively. The difference was statistically significant, p = 0.0002. No patient had detectable morbidity as a consequence of seed emboli. CONCLUSION: The use of linked seeds embedded in vicryl sutures for the peripheral portion of permanent radioactive seed prostate implants significantly reduced the incidence of pulmonary seed embolization in patients treated with the Seattle technique. PMID- 9869229 TI - Improved freedom from PSA failure with whole pelvic irradiation for high-risk prostate cancer. AB - PURPOSE: To determine the impact of whole pelvic irradiation on the risk of PSA failure in prostate cancer patients, at high predicted risk for lymph node involvement, receiving definitive radiotherapy. MATERIALS AND METHODS: Between October 1987 and December 1995, 506 patients with clinically localized prostate cancer were treated with definitive radiotherapy at UCSF and affiliated institutions. Treatment consisted of 4-field whole pelvic irradiation followed by a prostate-only boost, or prostate-only treatment (median follow-up was 35 months and 30 months, respectively). PSA failure was defined as: 1. a PSA value > or = 1 ng/ml; or 2. a PSA value that rose > or = 0.5 ng/ml in < or = 1 year posttreatment on two consecutive measurements, with the first rise defined as the time of failure. The calculated risk of lymph node positivity (%rLN+) was defined as 2/3(iPSA) + 10(GS-6), and high risk was defined as %rLN+ > or = 15%. Univariate and multivariate analyses were performed. RESULTS: A total of 201 high risk patients were identified. High-risk patients who received whole pelvic irradiation had significantly improved freedom from PSA failure compared to those who received prostate-only treatment (median PFS = 34.3 months vs. 21.0 months; p = 0.0001). Potential confounding variables, including initial PSA, Gleason score, T stage, radiation dose, year of treatment, use of three-dimensional (3D) conformal techniques, and use of hormone therapy, did not account for the observed difference in time to PSA failure. Multivariate analysis revealed type of radiation treatment to be the most significant independent predictor of outcome. CONCLUSION: Whole pelvic radiotherapy significantly improves the PSA failure-free survival in patients with a high calculated risk of lymph node positivity. PMID- 9869231 TI - Anemia is associated with decreased survival and increased locoregional failure in patients with locally advanced head and neck carcinoma: a secondary analysis of RTOG 85-27. AB - PURPOSE: The purpose of the present study is to investigate the strength of association between anemia and overall survival, locoregional failure, and late radiation therapy (RT) complications in a large prospective study of patients with advanced head and neck cancer treated with conventional radiotherapy with or without a hypoxic cell sensitizer. METHODS AND MATERIALS: Between March 1988 and September 1991, 521 patients with Stage III or IV squamous cell carcinoma of the head and neck were entered into a randomized trial examining the addition of etanidazole (SR 2508) to conventional radiation therapy (RT) (66-74 Gy in 33-37 fractions, 5 days a week). Patients with hemoglobin (Hgb) levels measured and recorded prior to the second week of RT were included in this secondary analysis. Hemoglobin levels were stratified as normal (> or = 14.5 gm% for men, > or = 13 gm% for women) or anemic (< 14.5 gm% for men, < 13 gm% for women). Locoregional failure rates were calculated using the cumulative incidence approach. Overall survival was estimated according to the Kaplan-Meier method. Late RT toxicity was scored according to the RTOG morbidity scale. Differences in rates of overall survival, locoregional failure, and late complications were tested by the Cox proportional hazard model. RESULTS: Of 504 eligible patients, 451 had a Hgb level measured and recorded prior to the second week of RT. One hundred sixty-two patients (35.9%) were considered to have a normal Hgb level and 289 patients (64.1%) were considered to be anemic. The estimated survival rate is 35.7% at 5 years in patients with a normal Hgb, versus 21.7% in anemic patients (p = 0.0016). The estimated locoregional failure rate is 51.6% at 5 years in patients with a normal Hgb, versus 67.8% in anemic patients (p = 0.00028). The estimated rate of grade 3 or greater toxicity is 19.8% at 5 years in patients with a normal Hgb, versus 12.7% in anemic patients (p = 0.063). On multivariate analysis, several variables were found to be independent predictors of survival including: T stage, Karnofsky performance status, N stage, age, total radiation dose to the primary, and Hgb level. Independent predictors of locoregional control included T stage, Karnofsky performance status, N stage, radiation dose, and Hgb level. The only variables which predicted for the development of late RT complications were gender (p = 0.0109) and age (p = 0.0167). These findings were consistent regardless of whether Hgb level was considered a dichotomous or continuous variable. CONCLUSION: Low Hgb levels are associated with a statistically significant reduction in survival and an increase in locoregional failure in this large prospective study of patients with advanced head and neck cancer. Hgb level should be considered as a stratification variable in subsequent studies of head and neck cancer. Strategies to increase Hgb prior to RT in patients with head and neck cancer may lead to improved survival and loco-regional control. PMID- 9869232 TI - Carcinoma of the tonsillar fossa: prognostic factors and long-term therapy outcome. AB - PURPOSE: To identify prognostic parameters and evaluate the therapeutic outcomes for patients with carcinoma of the tonsillar fossa treated with three treatment modalities. METHODS AND MATERIALS: The results of therapy are reported in 384 patients with histologically proven epidermoid carcinoma of the tonsillar fossa; 154 were treated with irradiation alone (55-70 Gy), 144 with preoperative radiation therapy (20-40 Gy), and 86 with postoperative irradiation (50-60 Gy). The operation in all but four patients in the last two groups consisted of an en bloc radical tonsillectomy with ipsilateral lymph node dissection. RESULTS: Treatment modality and total irradiation doses had no impact on survival. Actuarial 10-year disease-free survival rates were 65% for patients with T1 tumors, 60% for T2, 60% for T3, and 30% for T4 disease. Patients with no cervical lymphadenopathy or with a small metastatic lymph node (N1) had better disease free survival (60% and 70%, respectively) at 5 years than those with large or fixed lymph nodes (30%). Primary tumor recurrence (local, marginal) rates in the T1, T2, and T3 groups were 20-25% in patients treated with irradiation and surgery and 31% for those treated with irradiation alone (difference not statistically significant). In patients with T4 disease treated with surgery and postoperative irradiation, the local failure rate was 32% compared with 86% with low-dose preoperative irradiation and 47% with irradiation alone (p = 0.03). The overall recurrence rates in the neck were 10% for N0 patients, 25% for N1 and N2, and 35-40% for patients with N3 cervical lymph nodes, without significant differences among the various treatment groups. The incidence of contralateral neck recurrences was 8% with the various treatment modalities. On multivariate analysis the only significant factors for local tumor control and disease-free survival were T and N stage (p = 0.04-0.001). Fatal complications were noted in 7 of 144 (5%) patients treated with preoperative irradiation and surgery, 2 of 86 (2%) of those receiving postoperative irradiation, and 2 of 154 (1.3%) patients treated with radiation therapy alone. Other moderate or severe nonfatal sequelae were noted in 30% of the patients treated with preoperative irradiation and surgery, in 53% treated with postoperative irradiation, and in 19% receiving radiation therapy alone. CONCLUSION: Primary tumor and neck node stage are the only significant prognostic factors influencing locoregional tumor control and disease-free survival. Treatment modality had no significant impact on outcome. Radiation therapy remains the treatment of choice for patients with stage T1-T2 carcinoma of the tonsillar fossa. In patients with T3-T4 tumors and good general condition, combination surgery and postoperative irradiation offers better tumor control than single-modality and preoperative irradiation procedures, but with greater morbidity. PMID- 9869233 TI - Intraoperative electron beam radiotherapy for previously irradiated advanced head and neck malignancies. AB - PURPOSE: This is a retrospective review to evaluate the role of surgery and intraoperative electron beam radiotherapy (IOERT) in the treatment of patients with previously irradiated advanced head and neck cancers. METHODS AND MATERIALS: Between January 1992 and March 1997, 38 patients (31 males, 7 females; median age of 62 years) with recurrent head and neck cancer were treated with maximal resection and IOERT at the Ohio State University (OSU). All had been previously treated with full-course radiotherapy (median 65.1 Gy, range 50-74.4 Gy). Twenty nine patients (76%) had previously undergone one or more surgical procedures. After maximal surgery the tumor bed was treated with IOERT (single field in 36 patients and 2 fields in 2 patients), most commonly with 6 MeV electrons (87%). The dose administered (at 90% isodose line) was 15 Gy for close or microscopically positive margins in 34 patients and 20 Gy for gross disease in 1 patient. Further external beam radiation therapy (EBRT) was not given. RESULTS: After a median follow-up of 30 months (range 8-39 months), 24 of the 38 patients (66%) recurred within the IOERT field. Median time to IOERT failure was 6 months (95% CI: 4.3-7.7). The 6-month, 1-, and 2-year control rates within the IOERT volume were 41%, 19%, and 13%, respectively. Thirty of the 38 patients (79%) recurred in locoregional areas. Median time to locoregional failure was 4 months (95% CI: 3.3-4.7). The 6-month, 1-, and 2-year locoregional control rates were 33%, 11%, and 4%, respectively. Distant metastases occurred in 7 patients, 5 in association with IOERT failure and 2 with locoregional failure. Median overall survival was 7 months (95% CI: 4.7-9.3). The 6-month, 1-, 2-, and 3-year actuarial survival rates were 51%, 21%, 21%, and 8%, respectively. Major treatment-related complications occurred in 6 patients (16%). CONCLUSION: IOERT alone, at the dose used, is not sufficient for control of recurrent, previously irradiated head and neck cancers. Since higher IOERT doses are associated with high morbidity, we are currently evaluating the addition of limited EBRT dose and/or brachytherapy to improve the local control of these poor prognostic recurrent tumors, with acceptable morbidity. PMID- 9869234 TI - Concurrent radiochemotherapy for patients with stage III non-small-cell lung cancer (NSCLC): long-term results of a phase II study. AB - PURPOSE: To investigate the feasibility and activity of concurrent radiochemotherapy in patients with Stage III nonsmall-cell lung cancer (NSCLC). MATERIALS AND METHODS: Forty-one patients were treated with hyperfractionated radiation therapy (HfxRT) using 1.2 Gy bid, to a total of 69.6 Gy and concurrent low-dose daily chemotherapy (CHT) consisting of 30 mg of carboplatin (CBDCA) and 30 mg of etoposide (VP-16) given Mondays to Fridays during the RT course. On Saturdays and Sundays during the RT course, CBDCA and VP-16 were both given in a daily dose of 100 mg each. RESULTS: Median survival time was 25 months, and 3- and 5-year survival rates were 34% and 29%, respectively. Median relapse-free survival time was 22 months, and 3- and 5-year relapse-free survival rates were 32%, and 29%, respectively. Median time to local recurrence was 24 months and 3- and 5-year local recurrence-free survival rates were 41% and 38%, respectively. Median time to distant metastasis was 28 months, and 3- and 5-year distant metastasis-free survival rates were 44% and 44%, respectively. Acute high-grade (> or = 3) toxicity was mostly hematological (30%), esophageal (15%), and bronchopulmonary (12%). Late high-grade toxicity was infrequent. CONCLUSION: This combined radiochemotherapy regimen produced promising results and warrants further studies with more patients before testing it in a prospective randomized fashion. PMID- 9869235 TI - A subgroup analysis of the Scripps Coronary Radiation to Inhibit Proliferation Poststenting Trial. AB - INTRODUCTION: In the Scripps Coronary Radiation to Inhibit Proliferation Poststenting (SCRIPPS) Trial, 192Ir significantly reduced angiographic, ultrasonographic, and clinical endpoints of restenosis. The objective of this analysis was to quantitate the impact of patient, lesion and technical characteristics on late angiographic outcome. METHODS: Patients with restenotic, stented coronary lesions were randomized to receive either 192Ir or placebo sources. Late luminal loss and loss index were calculated for several patient subgroups, including patients with diabetes, in-stent restenosis, multiple previous percutaneous transluminal coronary angioplasty (PTCA) procedures, longer lesion lengths, saphenous vein grafts, small vessel diameters, and minimum dose exposures < 8.00 Gy. Two-factor analysis of variance was used to test for an interaction between patient characteristics and treatment effect. RESULTS: In the treated group, late loss was particularly low in patients with diabetes (0.19 mm), in-stent restenosis (0.17 mm), reference vessel diameters < 3.0 mm (0.07 mm), and patients who received a minimum radiation dose to the entire adventitial border of at least 8.00 Gy. The loss index in each of these subgroups was similarly low at -0.02, 0.03, -0.02, and 0.03, respectively. By 2-factor analysis of variance, a significant interaction between subgroup characteristic and treatment effect (late loss) was found in patients with in-stent restenosis (p = 0.035), and patients receiving a minimum dose of 8.00 Gy to the adventitial border (p = 0.009). CONCLUSION: In this pilot study, patient characteristics associated with a more aggressive proliferative response to injury appeared to confer an enhanced response to radiotherapy. Furthermore, a dose threshold response to 192Ir was found with an enhanced response occurring when the entire circumference of the adventitial border was exposed to at least 8.00 Gy. PMID- 9869236 TI - Prevalence of mood disorders and utility of the PRIME-MD in patients undergoing radiation therapy. AB - PURPOSE: To validate a short, structured interview procedure that allows practicing oncologists to quickly and reliably identify mood disorders in their patients, and to estimate the prevalence and types of mood disorders in a radiation therapy patient setting, noting relationships between mood disorders and patient characteristics. METHODS: Consecutive, eligible adult patients from the practices of two radiation oncologists were administered the Primary Care Evaluation of Mental Disorders (PRIME-MD) by the treating physician. A subset of these patients was also evaluated with the SCID, administered by trained mental health care personnel. Agreement between the two instruments was examined using the kappa statistic. Prevalence of mood disorders was determined from the PRIME MD. The significance of relationships between patient characteristics and mood disorders was examined by chi-square and ANOVA analysis, and subsequently by multivariate logistic regression analysis. RESULTS: One hundred twenty-two patients were studied. Fifty-three of these were administered the SCID. Agreement between the two instruments was very good (kappa = 0.70). A diagnosis of a depressive or anxiety disorder by the PRIME-MD was made in 59 of the 122 patients (48%, 95% confidence interval = 39%, 58%). Multivariate analysis showed that a diagnosis of a depressive mood disorder was significantly related to pain intensity and prior history of depression. CONCLUSION: We have demonstrated the validity and feasibility of the PRIME-MD administered by oncologists in making diagnoses of mood disorders. The prevalence of mood disorders in our set of patients undergoing a course of RT was nearly 50%. Future studies should describe the natural history of these disorders, and determine optimal intervention strategies. PMID- 9869237 TI - Low-dose total body irradiation and G-CSF without hematopoietic stem cell support in the treatment of relapsed or refractory acute myelogenous leukemia (AML), or AML in second or subsequent remission. AB - PURPOSE: Patients with relapsed acute myelogenous leukemia (AML), who are not eligible for bone marrow transplantation, have a poor prognosis when treated with chemotherapy alone. Total body irradiation (TBI) is an effective modality against AML when used in doses of 1000-1400 cGy with hematopoietic stem cell support. We undertook a phase I study of TBI with granulocyte-colony-stimulating factor (G CSF) support, without stem cell support in patients with AML either in relapse or second or subsequent remission. METHODS AND MATERIALS: Patients with relapsed AML, or AML in second or subsequent remission were treated in a phase I study of TBI followed by G-CSF. The first dose level was 200 cGy. After the initial cohort of patients it was clear that patients with overt leukemia did not benefit from this treatment, and subsequent patients were required to be in remission at the time of TBI. RESULTS: Eleven patients were treated, 4 in overt relapse, and 7 in remission. 200 cGy was used in all, and dose escalation was not possible due to prolonged thrombocytopenia in all patients but one. Neutrophil recovery was adequate in those patients who remained in remission after TBI. Patients with overt leukemia had transient reduction in blast counts, but rapid recurrence of their leukemia. Patients treated in remission had short remissions, with the exception of one patient who is in remission 32 months after treatment. CONCLUSION: There is some antileukemic effect of TBI even at 200 cGy, though this dose appears to be too low to help a significant number of patients. If TBI is to be escalated without stem cell support, then a thrombopoietic agent will need to be used. PMID- 9869238 TI - Radiation therapy in the management of symptomatic bone metastases: the effect of total dose and histology on pain relief and response duration. AB - PURPOSE: In order to better define variables and factors that may influence the pain response to radiation, and to look for a radiation regimen that can assure the highest percentage and the longest duration of pain relief, we performed a prospective, although not randomized, study on patients with bone metastases from various primary sites. METHODS AND MATERIALS: From December 1988 to March 1994, 205 patients with a total of 255 solitary or multiple bone metastases from several primary tumors were treated in our radiotherapy center with palliative intent. Irradiation fields were treated with three main fractionation schedules: (1) Conventional fractionation: 40-46 Gy/20-23 fractions in 5-5.5 weeks; (2) Short course: 30-36 Gy/10-12 fractions in 2-2.3 weeks; (3) Fast course: 8-28 Gy/1 4 consecutive fractions. Pain intensity was self-assessed by patients using a visual analogic scale graduated from 0 (no pain) to 10 (the strongest pain one can experience). Analgesic requirement was assessed by using a five-point scale, scoring both analgesic strength and frequency (0 = no drug or occasional nonopioids; 1 = Nonopioids once daily; 2 = Nonopioids more than once daily; 3 = Mild opioids (oral codeine, pentazocine, etc.), once daily; 4 = Mild opioids more than once daily; 5 = Strong opioids (morphine, meperidine, etc.). Complete pain relief meant the achievement of a score < or = 2 in the pain scale or 0 in the analgesic requirement scale. Partial pain relief indicated a score of 3 to 4 or of 1 to 2 on the former and latter scale, respectively. RESULTS: Total pain relief (complete + partial) was observed in 195 (76%) sites, in 158 of which (62%) a complete response was obtained. Metastases from NSC lung tumors appeared to be the least responsive among all primary tumors, with 46% complete pain relief in comparison to 65% and 83% complete relief in breast (p = 0.04) and in prostate metastases (p = 0.002), respectively. A significant difference in pain relief was detected among the several ranges of total dose delivered to the painful metastases, with 81%, 65%, and 46% complete relief rates in the 40-46 Gy, 30-36 Gy (p = 0.03), and 8-28 Gy (p = 0.0001) dose ranges respectively. A straight correlation between total dose and complete pain relief was confirmed by the curve calculated by the logistic model which shows that doses of 30 Gy or more are necessary to achieve complete pain relief in 70% or more of bone metastases. This correlation holds also for the duration of pain control, as shown by the actuarial analysis of time to pain progression. Multivariate analyses, with complete pain relief and time to pain progression as endpoints show a highly significant effect of radiation dose (p = 0.0007) and performance status (p = 0.003), with lower rates of complete pain relief and shorter time to pain progression observed after smaller radiation total doses or higher Eastern Cooperative Oncology Group (ECOG) scores. CONCLUSION: Although single-dose or short course irradiation is an attractive treatment in reducing the number of multiple visits to radiotherapy departments for patients with painful bone metastases, it is nevertheless clear that aggressive protracted treatments seem to offer significant advantages especially for patients in whom the expected life span is not short. PMID- 9869239 TI - Clinical results of nonsurgical treatment for spinal metastases. AB - PURPOSE: In contrast with many analyses of surgical treatment for spinal metastases, there have been only a few recent well-documented publications assessing nonsurgical treatment. This paper is a study of the outcome of nonsurgical therapy for metastatic tumors of the spine. METHODS AND MATERIALS: One hundred and one patients with spinal metastases were treated with radiation therapy and/or chemotherapy without surgical intervention between 1990 and 1995, in prospective analysis, and had follow-up for more than 24 months. This study included 59 men and 42 women with a mean age of 61 years (range: 14 to 81). Mean follow-up periods were 11 months for patients dying of the disease and 53 months for the survivors. Neurological status, pain relief, functional improvement, and cumulative survival rate were assessed. RESULTS: Of the total treated, 67 patients (66%) were evaluated as being neurologically stable or improved after treatment. Pain relief was achieved in 67%, and 64% showed functional improvement. Primary lesion responsiveness to nonsurgical therapy influenced the survival, neurological recovery, pain control, and function. Neurological findings before therapy were useful in predicting ambulatory status after treatment. CONCLUSION: Nonsurgical treatment was often successful when primary tumors had responsiveness to radiation therapy and/or chemotherapy. We found this to be evident even when neurological deficits were found, particularly in lumbar spines. Spinal metastases of tumors with less responsiveness, unless patients were neurologically intact, responded poorly to therapy. Most of the patients who were successfully treated enjoyed relief lasting nearly until death. Their functional ability was limited by general debility, rather than by local tumor regeneration. PMID- 9869240 TI - Radiosensitivity of normal tissues in ataxia-telangiectasia heterozygotes. AB - PURPOSE: Approximately 5% of cancer patients given radiation therapy exhibit severe injuries to the noncancerous tissue in the radiation field. Striking clinical sensitivity to ionizing radiation has been observed frequently in ataxia telangiectasia (A-T) homozygotes. This study was undertaken to test the hypothesis that heterozygous carriers of a mutated gene for A-T may represent a substantial proportion of all patients who suffer severe radiation toxicity. METHODS: The medical records of all A-T heterozygotes treated with radiation therapy for breast or prostate cancer were compiled from an ongoing study of mortality and cancer incidence in A-T families. Diagnostic, treatment, and follow up records were reviewed. Acute and long-term radiation complications were scored according to Radiation Therapy and Oncology Group criteria. RESULTS: There were no instances of soft tissue necrosis or other apparent serious injuries to normal tissues of two A-T heterozygotes with prostate carcinoma and 11 with breast carcinoma who received moderate-to-high doses of conventionally fractionated radiation therapy by megavoltage techniques. CONCLUSION: There is no evidence that abnormal clinical radiosensitivity occurs in A-T heterozygotes receiving conventionally fractionated radiation therapy for breast or prostate cancer. PMID- 9869242 TI - Discrimination of human tumor radioresponsiveness using low-dose rate irradiation. AB - PURPOSE: Evaluation of the theoretical and practical value of using low-dose rate (LDR) irradiation to increase the resolution of radiosensitivity testing of primary human tumors using clonogenic assays. METHODS AND MATERIALS: Fourteen human tumor cell lines were assessed for surviving fraction at 2-8 Gy (SF2-SF8) using low-dose rate irradiation and a clonogenic assay. Further data were collected from the literature for 64 low-dose rate irradiation survival curves from human tumor cell lines. The data were grouped into five different radioresponsiveness categories (A-E). An analysis was made of the ability of the graded survival levels to discriminate between the different radioresponse groups and compared with previous analyses for high-dose rate SF2. Fifteen human cervical carcinoma specimens were analysed for SF2 and SF3.5 following high- and low-dose rate irradiation. RESULTS: Low-dose rate irradiation increased the spread of tumor cell line radiosensitivity data and the ability to discriminate between radioresponse groups was greater at low than at high-dose rates. Using low-dose rate irradiation on primary tumor specimens and a soft agar clonogenic assay decreased the success rate in obtaining data. The latter dropped from 70% for high-dose rate SF2 to 51% for low-dose rate SF3.5. CONCLUSIONS: The work on cell lines illustrates that low-dose rate irradiation does improve the ability of clonogenic radiosensitivity measurements to discriminate between tumors of different radioresponsiveness groups. However, using low-dose rate irradiation on primary human tumors with a soft agar clonogenic assay was not practical because of reducing the success rate for obtaining data for radiosensitivity measurements. PMID- 9869241 TI - Induction of transforming growth factor alpha in irradiated mouse jejunum. AB - PURPOSE: To determine the involvement of the mitogenic growth factors transforming growth factor alpha (TGF alpha), epidermal growth factor (EGF), and the EGF receptor (EGF-R) in the proliferative response after irradiation of the mouse jejunum. METHODS AND MATERIALS: C3Hf/Kam mice were whole-body irradiated with 5 and 11 Gy 250 kV X rays. Mice were killed 1-10 days after irradiation, and immunohistochemistry, in situ hybridization (ISH), and RNase protection assays were performed. RESULTS: Damage to the jejunal crypts caused by irradiation resulted in a strong proliferative response 1-5 days after 5 Gy and 3-6 days after 11 Gy. Expression of TGF alpha, EGF, and EGF-R increased at 1-2 days and decreased at 4-8 days after 5- or 11-Gy irradiation. Also, TGF alpha mRNA increased during the early phase of the proliferative response (1-2 days after 5 or 11 Gy) followed by a decrease at 4 days after 5 Gy and 8 days after 11 Gy. CONCLUSION: These data indicate that, at the beginning of the proliferative response after irradiation, the transcription of TGF alpha mRNA is increased, and that it is inhibited just before compensatory proliferation decreases. Thus, active regulation of TGF alpha expression takes place at least at the transcriptional level, resulting in upregulation of TGF alpha production and increased TGF alpha levels in the crypts during the proliferative response. PMID- 9869243 TI - Target position variability throughout prostate radiotherapy. AB - PURPOSE: To quantify the variability in prostate and seminal vesicle position during a course of external beam radiotherapy, and to measure the proportion of target variability due to setup error. METHODS AND MATERIALS: Forty-four weekly planning computerized tomography (CT) studies were obtained on six patients undergoing radiotherapy for prostate cancer. All patients were scanned in the radiotherapy treatment position, supine with an empty bladder, with no immobilization device. All organs were outlined on 3-mm-thick axial CT images. Anterior and lateral beam's eye view digitally reconstructed radiographs and multiplanar reformatted images were generated. The position of the prostate and seminal vesicles relative to the isocenter location as set that day was recorded for each CT study. Target position relative to a bony landmark was measured to determine the relative contribution of setup error to the target position variability. RESULTS: The seminal vesicle and prostate position variability was most significant in the anterior-posterior (AP) direction, followed by cranial caudal (CC) and mediolateral (ML) directions. Setup error contributed significantly to the total target position variability. Rectal filling was associated with a trend to anterior movement of the prostate, whereas bladder filling was not associated with any trends. Although most deviations from the target position determined at the initial planning CT scan were within 10 mm, deviations as large as 15 mm and 19 mm were seen in the prostate and seminal vesicles respectively. Target position variations were evenly distributed around the initial target position for some patient studies, but unpredictable patterns were also seen. From a simulation based on the observed variability in target position, the AP, CC, and ML planning target volume (PTV) borders around the clinical target volume (CTV) required for target coverage with 95% certainty are 12.4 mm, 10.3 mm, and 5.6 mm respectively for the prostate and 13.8 mm, 8.6 mm, and 3.9 mm respectively for the seminal vesicles. CONCLUSION: Target position variability is significant during prostate radiotherapy, requiring large PTV borders around the CTV. This target position variability may be potentially reduced by improving the setup accuracy. PMID- 9869244 TI - A change in treatment process with a modern record and verify system. AB - PURPOSE: With the introduction of new treatment devices, such as a multileaf collimator (MLC) and dynamic wedge (DW), therapists have an increased responsibility to ensure correct treatment. Simultaneously, three-dimensional treatment planning (3DTP) has led to an increased number of portals and table movements. To counteract this challenge and maintain efficiency, a comprehensive record and verify (R&V) system is mandatory. We evaluated a commercial system (Varis) for reliability, ease of use, efficiency, and integration with our planning systems. METHODS AND MATERIALS: Some key elements of the Varis system are: integration of MLC and DW; auto setup for MLC, jaw, collimator, gantry, and limited table parameters; direct download of simulation beam data; and a regimented field scheduling system that prescribes all beam data for particular fractions. Evaluation of the system was driven by treatment time analysis, error rates, and an increased workload. These issues were governed by how we disseminated duties and how the system accommodated or changed our processes. RESULTS: Most data entry is performed by our dosimetry staff. Data can be downloaded from the simulator, but more patients now move from CT simulation and/or 3DTP to the treatment machine. Varis does not link to these systems. The physics staff confirms all entries to correct data entry errors. The workload for dosimetrists increased by an average of 8 minutes/patient entry; physics time increased by 7 minutes/patient entry; the weekly electronic chart check takes approximately 3 minutes/patient. Therapists who used Varis efficiently showed a slight decrease in treatment times, attributed to MLC integration and auto-setup. Some therapists experienced a decrease in efficiency, because of unfamiliarity and excess intervention. On a positive note, notable events have decreased by a factor of 10 since full initiation. Unfortunately, the remaining errors are often the result of a therapist relying on incorrect electronic information. CONCLUSION: The Varis R&V system has had an impact on our clinic's process and efficiency. Checking of all beam data and related field scheduling have helped reduce errors and misconceptions. We feel a dual-energy machine can be operated with two experienced therapists and an up-to-date R&V system more accurately and efficiently than with three therapists working without an integrated R&V. We anticipate future Varis releases will further promote efficiency and accuracy. PMID- 9869245 TI - Volumetric and dosimetric evaluation of radiation treatment plans: radiation conformity index. AB - PURPOSE: The use of conformal radiation therapy has grown substantially during the last years since three-dimensional (3D) treatment planning systems with beams eye-view planning has become commercially available. We studied the degree of conformity reached in clinical routines for some common diagnoses treated at our department by calculating a radiation conformity index (RCI). METHODS AND MATERIALS: The radiation conformity index, determined as the ratio between the target volume (PTV) and the irradiated volume, has been evaluated for 57 patients treated with 3D treatment plans. RESULTS AND CONCLUSION: The RCI was found to vary from 0.3 to 0.6 (average 0.4), a surprisingly low figure. The higher RCI is typical for pelvic treatments (e.g., prostate) and stereotactic treatments. The lower RCI is found for extended tumors, such as mammary carcinomas where the adjacent nodes are included. The latter is also valid for most lung cancer patients studied. The RCI gives a consistent method for quantifying the degree of conformity based on isodose surfaces and volumes. Care during interpretation of RCI must always be taken, since small changes in the minimum dose can dramatically change the treated volume. PMID- 9869246 TI - Endowed professorships in radiation oncology. PMID- 9869247 TI - Pentoxifylline treatment of radiation mastitis. PMID- 9869248 TI - Re: Hsu et al. 1998; 40(2):405-410. PMID- 9869249 TI - Techniques for measurement and assessment of mobility in rehabilitation: a theoretical approach. AB - OBJECTIVE: Mobility is an important construct in rehabilitation; many instruments have emerged which measure or assess (aspects of) mobility. In the selection or development of an appropriate technique, knowledge about the fundamentals of rehabilitation medicine is needed, as well as about essential characteristics of techniques and fundamental differences between them. The aim of this paper is to classify, assess and discuss current techniques which are or can be used to measure aspects of mobility. DESIGN: Eight techniques (physical science techniques, clinimetry, observation, diaries, questionnaires, actigraphy, physiological techniques and activity monitors) are classified, assessed and discussed, based on the level of outcome measures, the aspect of mobility they measure, and methodological and practical criteria. RESULTS AND CONCLUSIONS: Rehabilitation medicine has a particular need for instruments that enable measurement of outcome measures on the level of activity and role fulfillment. Techniques differ in the type and number of mobility aspects they measure. Furthermore, important differences exist based on methodological and practical criteria. One optimal technique does not exist: the choice of a technique always depends on a complexity of factors, such as clinical problem, research questions, the mobility aspect of interest, required methodological strength, costs and availability. PMID- 9869250 TI - The impact of an information pack on patients with stroke and their carers: a randomized controlled trial. AB - OBJECTIVES: To assess the impact of information packs on patients with stroke and their carers, and to pilot some of the methodology for a trial of a Family Support Organiser (FSO). SUBJECTS: Seventy-one patients admitted to Oxford hospitals with acute stroke during February-July 1995, and 49 informal carers of these patients. DESIGN: Randomized controlled trial. Intervention group received an Information pack containing various Stroke Association publications one month after their stroke, or at discharge from hospital, whichever was sooner. Control group received nothing. Follow-up was by interview at the place of residence of the patients six months after their stroke. MEASURES: Outcome measures assessed knowledge about stroke; satisfaction with information received; patient behaviour in terms of access to community services and benefits; and health status and quality of life. RESULTS: Patients and carers in the intervention group tended to know more about stroke, but these differences were not significant once adjusted for age. Patients in the intervention group (but not carers) tended to be more satisfied with the information that they had received, but the differences were not significant. There were no differences with regard to any aspects of quality of life in patients in the intervention group, though carers in the intervention group were found to have significantly better mental health (p = 0.04). CONCLUSIONS: While the study was too small to generate firm conclusions, information leaflets may lead to improved knowledge about stroke several months after they have been distributed. This finding is worth following up with larger studies. The stroke knowledge questionnaire that was piloted in this trial seems to be able to detect differences between groups. PMID- 9869251 TI - A comparison of two physiotherapy treatment approaches to improve walking in multiple sclerosis: a pilot randomized controlled study. AB - OBJECTIVE: To use a pilot study to compare two physiotherapy approaches to improve walking in patients with gait disturbance due to multiple sclerosis (MS). DESIGN: Patients were assessed and then randomly assigned to one of two groups using a block randomization technique. They were treated by the research physiotherapist for a minimum of 15 treatments over a 5-7-week period and then reassessed by an independent therapist one week after treatment. SETTING: Both assessment and treatment were undertaken at a specialist rehabilitation centre. SUBJECTS: Outpatients with clinically stable MS (chronic progressive or relapsing remitting types) who were referred for physiotherapy to improve their mobility. INTERVENTION: Comparison was between a facilitation (impairment-based) approach and a task-oriented (disability-focused) approach. OUTCOME: Mobility was assessed using four measures: the 10-metre timed walk, the Rivermead Mobility Index, stride length and the Rivermead Visual Gait Assessment. Balance was assessed using the Berg Balance Test. RESULTS: Twenty-three patients were entered, and 10 in each group completed the study. The groups were similar on all measured items both before and after treatment. There was no significant difference in improvement between the two approaches. Following treatment, patients in both groups showed a significant overall improvement (p < 0.05) in both impairment and disability measures. CONCLUSIONS: No significant differences in effectiveness between the two methods were demonstrated. Both a task-oriented approach and a facilitation approach to the treatment of MS outpatients were associated with improved mobility. PMID- 9869252 TI - Predictive factors for the outcome of a multidisciplinary pain rehabilitation programme on sick-leave and life satisfaction in patients with whiplash trauma and other myofascial pain: a follow-up study. AB - OBJECTIVE: To evaluate the effects of a multidisciplinary rehabilitation programme on sick-leave, coping resources and life satisfaction in whiplash patients and other pain patients. SUBJECTS: Forty patients suffering from symptoms after whiplash trauma and 33 patients with musculoskeletal pain in the neck or back were recruited for this study. Ninety-seven consecutive patients admitted to the Department of Neurosurgery with cervical pain, cervical disc herniation or symptomatic spondylosis served as a control group. RESULTS: Decreased coping resources and poorer life satisfaction were observed for whiplash subjects at the beginning of the rehabilitation programme compared to the control group from the Department of Neurosurgery. After the rehabilitation period 49% of the patients had improved their coping resources totalling to 63% after 2 years. At that follow-up 46% of patients had increased their life satisfaction. Furthermore, the group with whiplash injury showed a significant increase in sick absenteeism whereas the group without whiplash trauma had decreased their sick-leave. Eighty-eight per cent of the subjects could be correctly classified according to their vocational outcome by means of discriminant function. The elapse of time since working, low life satisfaction, lack of increase in coping resources during the rehabilitation programme, ethnic origin of the patient and living in the countryside predicted poor vocational outcome. CONCLUSION: Our results suggest variables from the social environment and coping resources as useful predictors for treatment outcome. PMID- 9869253 TI - Comparison between two devices for measuring hip joint motions. AB - OBJECTIVE: To compare the reliability of measurements of hip motions obtained with two instruments, an electronic inclinometer and a two-arm goniometer, and to investigate whether the two instruments, and different body positions, produce the same measurement data. METHODS: Maximal active and passive hip movements were measured simultaneously with both instruments, in nine subjects during 10 consecutive measurements at short intervals. RESULTS: Intra-observer variability was lower with the inclinometer in measurements of passive hip rotations. The two instruments showed equal intra-observer variability for hip movements in general. The inclinometer showed lower inter-observer variability in the measurements of active internal rotation. More rotational movement was measured with the two-arm goniometer; more extension and flexion with the inclinometer. Also, more rotational movement was found in the prone position compared to sitting and supine positions. CONCLUSIONS: The inclinometer is more reliable in measurements of hip rotation. For hip movements in general the two-arm goniometer is just as accurate when used by only one observer. The two instruments, and some positions, are not interchangeable during consecutive measurements. PMID- 9869254 TI - The assessment of depression in aphasic stroke patients: the development of the Stroke Aphasic Depression Questionnaire. AB - OBJECTIVE: The Stroke Aphasic Depression Questionnaire (SADQ) was developed to detect depressed mood in aphasic patients in the community. DESIGN: Correlation analysis between new questionnaire and established measures. SETTING: Patients at home. METHODS: Seventy stroke patients who had been discharged from hospital were assessed on the SADQ, the Hospital Anxiety and Depression Scale and the Wakefield Depression Inventory. The SADQ was also administered to 17 aphasic patients on two occasions at a four-week interval. RESULTS: Results indicated that scores on the SADQ were significantly related to other measures of depression (r(s) = 0.22 0.52, p <0.05). A shortened 10-item version showed higher validity (r(s) = 0.32 0.67, p <0.01). Test-retest analysis indicated the SADQ is reliable over a four week interval (r(s) = 0.72, p <0.001). CONCLUSIONS: The SADQ may be used for assessing depressed mood in aphasic patients in the community though further validation is required. PMID- 9869256 TI - Measuring outcome in community-based rehabilitation services for people who have suffered traumatic brain injury: the Community Outcome Scale. AB - OBJECTIVE: To develop a way of measuring long-term outcomes after traumatic brain injury (TBI) that takes account of individual circumstances. DESIGN: Reports by head-injured people and their families about problems and coping strategies were elicited via semi-structured interviews. Specially designed computer software was used to record problems and strengths and to measure their impact for the individual on four handicap dimensions. SETTING: Interviews took place in the respondents' homes. SUBJECTS: Between 1992 and 1997 the services at 10 rehabilitation centres in England were evaluated. Subjects were men and women aged between 16 and 65 who had suffered TBI and who had been referred for treatment to one of the centres under study. INTERVENTIONS: Head-injured people and their families were interviewed three months, 18 months and in some cases 36 months after recruitment. MAIN OUTCOME MEASURES: The Hospital Anxiety and Depression Scale (HAD) and the Functional Independence/Assessment Measure (FIM/FAM) were used to validate relevant dimensions of the new scale. RESULTS: A measure of long-term outcome was developed and called the Community Outcome Scale. Relevant dimensions of the scale showed significant correlation with the HAD and with items of the FIM/FAM. CONCLUSION: The scale captures information which no other instrument does, and merits further validation. PMID- 9869255 TI - Cognitive deficits following stroke in the basal ganglia. AB - OBJECTIVE: To examine the effect of a stroke in the basal ganglia on cognitive functioning. DESIGN: As part of a larger prospective study on the neuropsychological and psychosocial consequences of stroke, 12 patients with a stroke confined to the basal ganglia were examined. SETTING: The patients were assessed in one of the three participating hospitals. SUBJECTS: The results of 12 patients with a stroke in the basal ganglia (mean age 55 years), were compared to 24 controls (mean age 54.3 years). MAIN OUTCOME MEASURES: Different aspects of memory, attention and concentration, visuospatial and visuoconstructive functions, language and arithmetic were assessed using neuropsychological tests. RESULTS: The data show a significant abnormality in cognitive functioning on all cognitive domains. CONCLUSIONS: The results stress the notion that subcortical damage may lead to substantial neuropsychological disorders and have clear implications for neurological rehabilitation. PMID- 9869257 TI - Memory facilitation and stimulation of endogenous nerve growth factor synthesis by the acetylcholine releaser PG-9. AB - The effects of PG-9 (3alpha-tropyl 2-(p-bromophenyl)propionate), the acetylcholine releaser, on memory processes and nerve growth factor (NGF) synthesis were evaluated. In the mouse passive-avoidance test, PG-9 (10-30 mg/kg, i.p.), administered 20 min before the training session, prevented amnesia induced by both the non selective antimuscarinic drug scopolamine and the M1-selective antagonist S-(-)-ET-126. In the same experimental conditions, PG-9 (5-20 microg per mouse, i.c.v.) was also able to prevent antimuscarine-induced amnesia, demonstrating a central localization of the activity. At the highest effective doses, PG-9 did not produce any collateral symptoms as revealed by the Irwin test, and it did not modify spontaneous motility and inspection activity, as revealed by the hole-board test. PG-9 was also able to increase the amount of NGF secreted in vitro by astrocytes in a dose-dependent manner. The maximal NGF contents obtained by PG-9 were 17.6-fold of the control value. During culture, no morphological changes were found at effective concentrations of PG-9. The current work indicates the ability of PG-9 to induce beneficial effects on cognitive processes and stimulate activity of NGF synthesis in astroglial cells. Therefore, PG-9 could represent a potential useful drug able to improve the function of impaired cognitive processes. PMID- 9869258 TI - Serotonin-independent model of cisplatin-induced emesis in the ferret. AB - para-Chlorophenylalanine (PCPA, 100-200 mg/kg) was used as a pharmacological tool to characterize the 5-hydroxytryptamine (5-HT) involvement in the emesis occurring 24 hr after the administration of cisplatin (10 mg/kg) in the ferret. PCPA was effective to antagonize the initial 8 hr period of retching and vomiting, but potentiated the emesis that occurred during the remaining 8- to 24 hr observation period. Tissue samples removed from the brainstem at 24 hr post injection of cisplatin alone revealed an elevation of 5-HT, dopamine and homovanillic acid that was antagonized by the injection of PCPA. Cisplatin also induced increases in the urinary levels of 5-hydroxyindoleacetic acid that was similarly antagonized by PCPA. Results are discussed in terms of the relevance of 5-HT to the model of cisplatin (10 mg/kg)-induced emesis in the ferret compared to the problem of acute and delayed emesis in man. The residual or delayed phase of cisplatin-induced emesis may involve a 5-HT-independent mechanism. PMID- 9869259 TI - Effects of KRN4884 (a novel K+ channel opener), levcromakalim, nilvadipine and propranolol on endothelin-1-induced heart disorders in anesthetized rats. AB - The effects of KRN4884 (5-amino-N-[2-(2-chrolophenyl)ethyl]-N'-cyano-3 pyridinecarboxa midine), a novel K+ channel opener, on the electrocardiogram changes caused by the intracoronary administration of endothelin-1 (ET-1) were studied in anesthetized rats and compared with the effects of levcromakalim, a K+ channel opener; nilvadipine, a Ca2+ antagonist; and propranolol, a beta adrenoceptor antagonist. KRN4884 (50 microg/kg, i.v.) and levcromakalim (300 microg/kg, i.v.) inhibited the ST segment elevation and the development of arrhythmias induced by ET-1 (5 microg, i.c.) and decreased the incidence of death. Nilvadipine (300 microg/kg, i.v.) and propranolol (1000 and 3000 microg/kg, i.v.) each prevented the ST segment elevation, but the suppressions of the occurrence of arrhythmias produced by nilvadipine and propranolol were less than that shown by KRN4884. KRN4884 (30 and 50 microg/kg, i.v.), levcromakalim (100 and 300 microg/kg, i.v.) and nilvadipine (100 and 300 microg/kg, i.v.) significantly decreased the mean blood pressure in a dose-dependent manner, but propranolol did not. The heart rate was decreased by nilvadipine (100 and 300 microg/kg, i.v.) and propranolol (1000 and 3000 microg/kg, i.v.), but was not affected by KRN4884 (30 and 50 microg/kg, i.v.) or levcromakalim (100 and 300 microg/kg, i.v.). These results suggest that pretreatments with KRN4884 and levcromakalim are more effective on ET-1-induced electrocardiogram changes than those with nilvadipine and propranolol. PMID- 9869260 TI - Effects of YT-146 [2-(1-octynyl) adenosine], an adenosine A2A receptor agonist, on cAMP production and noradrenaline release in PC12 cells. AB - Effects of YT-146 [2-(1-octynyl) adenosine], an adenosine A2 receptor agonist, on cAMP production and noradrenaline (NA) release were investigated in PC12 cells. YT-146 caused a concentration-dependent cAMP accumulation (EC50: 1.2+/-0.9 nM). In [3H]NA-prelabeled cells, YT-146 increased the basal NA release and enhanced ATP-evoked NA release in a concentration-dependent manner (EC50: 0.23+/-0.15 nM). YT-146 augmented the maximal response to ATP without affecting the EC50 value of ATP. These effects of YT-146 were inhibited by several adenosine receptor antagonists with a characteristic of adenosine A2A receptor subtype. The effects of YT-146 were mimicked by forskolin, dibutylyl cAMP and Sp-cAMPS, and inhibited by H-89, a cAMP-dependent protein kinase inhibitor. YT-146 had little effect on ATP-induced increase in intracellular Ca2+ concentration. YT-146 enhanced the NA release induced by several different stimuli including Ca2+ ionophore A23187. The present results suggest that YT-146 is a potent agonist on adenosine A2A receptors in PC12 cells and causes a cAMP-dependent enhancement of NA release by affecting the exocytosis process at a point downstream of the intracellular Ca2+ increase. PMID- 9869261 TI - Salivary amylase activity of rats fed a low calcium diet. AB - Wistar strain rats fed low calcium diets (1, 2, 3, 4, 6 and 8 weeks) exhibited changes in secretory function of whole saliva. In particular, there were changes in salivary flow rate, total salivary protein, amylase enzyme activity, salivary amylase content and the level of cyclic AMP in the parotid gland acinar cell. Although there were no changes for the first 3 weeks, the levels increased at week 4 and decreased at week 6. The wet weight of the parotid gland started to decrease at week 4. These results suggest that when fed low calcium diets for long periods of time, rats develop defective salivary secretion. PMID- 9869262 TI - Role of glutathione in the antiulcer effect of hot water extract of black tea (Camellia sinensis). AB - The role of a hot water extract of black tea (Camellia sinensis (L). O. Kuntze Theaceae) in the gastric cytoprotective mechanisms was studied using gastric mucosal lesions produced by various ulcerogens in rats as an experimental model. Prior oral administration of black tea extract (BTE) at 20 ml/kg, i.g. once a day for 7 days significantly reduced the incidence of gastric erosions and severity induced by ethanol, diethyldithiocarbamate (DDC) and diethylmaleate (DEM). This treatment also favorably altered the changes in acid and peptic activity of gastric juice in these ulcerogen-treated animals. Singular administration of succimer (60 mg/kg, i.g.), the standard sulfhydryl containing antiulcer drug used as a reference drug, was also effective. The levels of glutathione and glutathione peroxidase were significantly decreased after treatment with ethanol, DDC and DEM, and this decrease was prevented by BTE pretreatment in the aforesaid manner. Other major features of BTE-induced reversal of ulcerogenic agents include a significant decrease in the protein content and a marked increase in hexosamine and sialic acid content. These results suggest a major role for glutathione, an endogenous antioxidant, in the cytoprotection against ulceration afforded by BTE. PMID- 9869264 TI - Effects of monatepil maleate, a new Ca2+ channel antagonist with alpha1 adrenoceptor antagonistic activity, on cholesterol absorption and catabolism in high cholesterol diet-fed rabbits. AB - The mechanism of the prophylactic effect against hyperlipidemia by monatepil maleate was investigated in animal models. Monatepil maleate is an antihypertensive agent with Ca2+-channel antagonistic, alpha1-adrenergic receptor blocking, and lipid peroxidation inhibitory activity. In high cholesterol diet fed rabbits, monatepil maleate (30 mg/kg, p.o., once daily for 9 weeks) showed a prophylactic effect against increases in total cholesterol and beta-lipoprotein. Monatepil maleate significantly accelerated the clearance of radioactivity from the blood after intravenous injection of low-density lipoprotein (LDL) labeled with [1alpha,2alpha (n)-3H]cholesterol, increasing biliary excretion of [3H]-bile acids without modifying bile acid composition. Furthermore, monatepil maleate tended to inhibit the absorption of orally administered [1alpha,2alpha (n) 3H]cholesterol from the gastrointestinal tract in these rabbits. In Watanabe heritable hyperlipidemic (WHHL) rabbits, an animal model of hepatic LDL receptor deficiency, monatepil maleate (30 mg/kg, p.o., once daily for 6 months) did not suppress the increase in plasma lipids. These results suggest that the plasma lipid lowering effect of monatepil maleate requires the presence of hepatic LDL receptors. It is also suggested that monatepil maleate improves plasma lipid metabolism through two mechanisms: enhancement of clearance of plasma LDL, which may be mediated by up-regulation of hepatic LDL receptors, and acceleration of conversion of free cholesterol to bile acids in the liver. PMID- 9869263 TI - Decrease in participation of nitric oxide in nonadrenergic, noncholinergic relaxation of rat intestine with age. AB - Participation of nitric oxide in the electrical field stimulation-induced nonadrenergic, noncholinergic (NANC) relaxation in various intestinal regions was studied in 2- to 50-week-old Wistar rats. In the jejunum of 2-week-old rats, the extent of the nitric oxide-mediated component of the relaxation of longitudinal muscle was approximately 60-70%, whereas the component was 40-50% in 4-week-old rats and was absent in 8- and 50-week-old rats. Thus, nitric oxide seems to be the most important mediator at young ages but its significance is lost with age. The same tendency as that in the jejunum was also shown in longitudinal muscle of the ileum, proximal and distal colon, and rectum. The tendency was also shown in the circular muscle of the rectum. Sensitivity of the longitudinal muscle of the jejunum and proximal colon to exogenously added nitric oxide was high in younger rats. Immunoreactive structures for nitric oxide synthase were observed in the circular muscle layer of the rectum. The population of the structures was denser in 4-week-old than that in 50-week-old. The results suggest that NANC relaxation in every region of the intestine at 2-week-old is almost solely mediated by nitric oxide, and its significance as an inhibitory mediator gradually or rapidly decreases with age. PMID- 9869265 TI - Histamine H2-receptor antagonism of T-593, an anti-ulcer agent: studies on aminopyrine accumulation in isolated canine gastric mucosal cells. AB - Histamine H2-receptor antagonistic properties of the anti-ulcer agent T-593, (+/ )-(E)-1-[2-hydroxy-2-(4-hydroxyphenyl)ethyl] -3-[2[[[5-(methylamino)methyl-2 furyl]methyl]thio] ethyl] -2-(methylsulfonyl)guanidine, were investigated on [14C]aminopyrine accumulation in isolated canine gastric mucosal cells and compared with those of ranitidine and famotidine. The potency of T-593-inhibition of [14C]aminopyrine accumulation stimulated by 10(-4) M histamine, with an IC50 value of 1.85 x 10(-6) M, was approximately 5 times greater than that of ranitidine, but half that of famotidine. T-593 did not affect [14C]aminopyrine accumulation stimulated by carbachol or dibutyryl-cAMP. T-593 depressed the maximal response of the histamine concentration-response curve with a dose related displacement to the right, indicating that the nature of the H2-receptor antagonism of T-593 was insurmountable and included non-competitive inhibition. The inhibitory efficacy of T-593 was time-dependent and was retained after the cells were washed. The inhibitory potency of (-)-S-T-593, one of the enantiomers, on the [14C]aminopyrine accumulation stimulated by histamine was approximately twice that of racemic T-593 and it also behaved as an insurmountable H2-receptor antagonist. However, the potency of (+)-R-T-593 was markedly weak. These results suggest that T-593 has H2-receptor antagonism that is insurmountable and this agent slowly associates and dissociates with the receptor in isolated canine gastric mucosal cells and that the specific substance causing H2-receptor antagonism is (-)-S-T-593. PMID- 9869266 TI - Norepinephrine- and K+-induced Mn2+-dependent contractions and the dynamics of intracellular Mn2+ changes in dispersed smooth muscle cells from Ca2+-depleted Mn2+-loaded vas deferens of the guinea pig. AB - The cellular mechanisms of norepinephrine (NE)-induced Mn2+-dependent contractions were investigated in dispersed smooth muscle cells from guinea pig vas deferens by characterizing the effects of NE and K+ on cell length observed by videotape microscopy and on intracellular Mn2+ and Ca2+ concentration ([Mn2+]i and [Ca2+]i) observed by confocal microscopy. Both stimulants induced slow sustained contraction in Ca2+-depleted, Mn2+-accumulated cells (Mn2+-loaded cells), whereas they induced biphasic contractions in normal cells that were neither Ca2+-depleted nor Mn2+-loaded. In both conditions, the number of cells responding to NE as well as the magnitude of NE-induced contractions increased in a dose-dependent manner. Contractions induced by K+ in Mn2+-loaded strip preparations were markedly smaller than those induced by NE. Although individual K+-induced contractions in responsive Mn2+-loaded cells were as large as those induced by NE, a much smaller percentage of Mn2+-loaded cells was responsive to K+ than to NE. These results are consistent with the idea that the contractions of strip preparations may reflect the magnitude of the contractions of individual cells as well as the percentage of responsive cells in the preparations. Inconsistent with the contractions, the [Mn2+]i rise induced by K+ was larger than that induced by NE, and the percentage of cells responsive to K+ was larger than that responsive to NE. These results suggest that NE may increase the Mn2+ sensitivity of contractile elements. PMID- 9869267 TI - Tamsulosin: alpha1-adrenoceptor subtype-selectivity and comparison with terazosin. AB - Selectivity of tamsulosin and terazosin to functional alpha1-adrenoceptors was examined. Both drugs competitively inhibited the contractile responses to noradrenaline in different tissues where the responses were mediated through the alpha1D-, alpha1B- or alpha1L-subtype. Together with the affinities obtained in the binding study with cloned (alpha1a, alpha1b, alpha1d) and native (alpha1A and alpha1B) subtypes, the selectivity of tamsulosin was alpha1A>alpha1L, alpha1D>alpha1B. Terazosin had lower affinity at various subtypes than tamsulosin, but showed relatively high selectivity to the alpha1D-subtype. In the human prostate, tamsulosin was more than 30-fold higher in affinity than terazosin in functional and binding studies. PMID- 9869268 TI - Itch-associated response and antinociception induced by intracisternal endomorphins in mice. AB - Endomorphin-1 and endomorphin-2 are newly identified endogenous peptides and have high affinity and selectivity for mu-opioid receptors. The present experiments were conducted to determine whether intracisternal injection of these peptides would produce an itch-associated response and antinociception and to compare their effects to that of morphine. Endomorphin-1 and endomorphin-2 (0.3-3 nmol/mouse) elicited facial scratching characterized by bell-shaped dose-response curves with a peak effect at endomorphin-1 at 0.3 nmol/mouse and endomorphin-2 at 1 nmol/mouse. Their peak effects were inhibited by subcutaneous pretreatment with naloxone (1 mg/kg). Morphine (0.3-30 nmol/mouse) produced facial scratching, and its dose-response curve was also bell-shaped. Scratching of the body trunk, head and ears were not elicited by these doses of endomorphins and morphine. Endomorphin-1 and -2 at doses of 0.3-3 nmol/mouse produced dose-dependent antinociception, as measured with the tail-pressure test. The potency and duration of actions of these peptides were comparable to those of morphine. The results suggest that endomorphin-1 and endomorphin-2 are involved in itch signaling and pain-inhibiting functions of the brain. PMID- 9869269 TI - Effects of adrenergic and nitrergic blockade on theophylline-induced increase in peripheral blood flow in rat ear. AB - A bolus injection of theophylline produced a significant increase in peripheral blood flow in anesthetized rat ear, monitored by laser-Doppler flowmetry, with increases in arterial blood pressure and heart rate. These effects were attenuated by previous treatment with reserpine, but reserpine had no effect on the blood flow increase produced by acetylcholine. A dose of propranolol, which caused attenuation of the theophylline-induced increase in heart rate, did not change the peripheral blood flow. The higher dose of propranolol, which nearly canceled the increases in blood pressure and heart rate, caused attenuation of the blood flow increase but did not cancel it. However, the theophylline-induced flow increase was completely reversed by a nitric oxide synthase inhibitor, NG nitro-L-arginine methyl ester, which alone had no effect, without any change in arterial blood pressure and heart rate. Treatment of the rats with the dose of inhibitor slightly and significantly reduced the response of peripheral blood flow to acetylcholine. The other isomer, NG-nitro-D-arginine methyl ester, and the other inhibitor, NG-monomethyl-L-arginine, did not have such an effect. These results suggest that the flow increase is due to an independent effect on the heart with modification by autonomic reflexes and involves the adrenergic and nitrergic pathways. PMID- 9869270 TI - Cholesterol-lowering effects of NTE-122, a novel acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor, on cholesterol diet-fed rats and rabbits. AB - Pharmacological characterization of NTE-122 (trans-1,4-bis[[1-cyclohexyl-3-(4 dimethylamino phenyl)ureido]methyl]cyclohexane), a novel acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor, was performed with both in vitro and in vivo assay systems. NTE-122 inhibited microsomal ACAT activities of various tissues (liver of rabbit and rat, small intestine of rabbit and rat, and aorta of rabbit) and cultured cells (HepG2 and CaCo-2), with IC50 values from 1.2 to 9.6 nM. The inhibition mode of NTE-122 was competitive for HepG2 ACAT. NTE-122 had no effect on other lipid metabolizing enzymes, such as 3-hydroxy-3-methylglutaryl-CoA reductase, acyl-CoA synthetase, cholesterol esterase, lecithin:cholesterol acyltransferase, acyl-CoA:sn-glycerol-3-phosphate acyltransferase and cholesterol 7alpha-hydroxylase up to 10 microM. When NTE-122 was administered to the cholesterol diet-fed rats, serum and liver cholesterol levels were markedly reduced with an ED50 of 0.12 and 0.44 mg/kg/day, respectively. In the cholesterol diet-fed rabbits, NTE-122 significantly lowered plasma and liver cholesterol levels at more than 2 mg/kg/day. These results indicate that NTE-122 is a potent, selective and competitive inhibitor of ACAT, making it a worth while therapeutic agent for hypercholesterolemia and atherosclerosis. PMID- 9869271 TI - Profile of JTE-522 as a human cyclooxygenase-2 inhibitor. AB - Inhibitory activity and the mechanism of action of JTE-522 (4-(4-cyclohexyl-2 methyloxazol-5-yl)-2-fluorobenzenesulfonamid e), a novel selective cyclooxygenase (COX)-2 inhibitor, on human COX-1 and COX-2 were investigated and compared with those of reference compounds. In an enzyme assay, JTE-522 inhibited yeast expressed human recombinant COX-2 with an IC50 value of 0.085 microM. In contrast, JTE-522 did not inhibit human COX-1 prepared from human platelets at concentrations up to 100 microM. In a cell-based assay, JTE-522 diminished lipopolysaccharide-induced prostaglandin E2 production in human peripheral blood mononuclear cells (COX-2) (IC50 value = 15.1 nM). On the other hand, JTE-522 was less potent at inhibiting calcium ionophore-induced thromboxane B2 production in washed human platelets (COX-1) (IC50 value = 6210 nM). JTE-522 showed highly selective inhibition of human COX-2, and its activity was more selective than that of other COX-2 inhibitors (NS-398 and SC-58635). Human recombinant COX-2 activity was attenuated by JTE-522 in a dose-dependent and time-dependent manner. In contrast, the inhibitory activity of JTE-522 on human COX-1 was not affected by preincubation time. COX-2 inhibition by JTE-522 could not be recovered by gel filtration. These results indicate that JTE-522 is a highly selective, time dependent and irreversible inhibitor of human COX-2. PMID- 9869272 TI - (+/-)-cis-2-methylspiro[1,3-oxathiolane-5,3'-quinuclidine] hydrochloride, hemihydrate (SNI-2011, cevimeline hydrochloride) induces saliva and tear secretions in rats and mice: the role of muscarinic acetylcholine receptors. AB - We investigated effects of (+/-)-cis-2-methylspiro[1,3-oxathiolane-5,3' quinuclidine] hydrochloride, hemihydrate (SNI-2011, cevimeline hydrochloride), a rigid analogue of acetylcholine, on saliva and tear secretions in rats and mice to evaluate its therapeutical efficacy for xerostomia and xerophthalmia in patients with Sjogren's syndrome and X-ray exposure in the head and neck. Intraduodenal administrations of SNI-2011 increased saliva secretion in a dose dependent manner at doses ranging from 3 to 30 mg/kg in normal rats and mice, two strains of autoimmune disease mice and X-irradiated saliva secretion defective rats. The salivation elicited by SNI-2011 was completely inhibited by atropine. A similar atropine-sensitive response was observed in tear secretion. In rat submandibular/sublingual gland membranes, [3H]quinuclidinyl benzilate (QNB) binding was saturable, and Scatchard plot analysis revealed a single population of binding sites with a Kd of 22 pM and a maximal binding capacity of 60 fmol/mg protein. The competitive inhibition curve of the [3H]QNB binding by SNI-2011 was obtained, and its dissociation constant value calculated from IC50 was 1-2 microM. These results suggest that SNI-2011 increases saliva and tear secretions through a direct stimulation to muscarinic receptors in salivary and lacrimal glands, and they suggest that SNI-2011 should be beneficial to patients with Sjogren's syndrome and X-ray exposure in the head and neck. PMID- 9869273 TI - Comparative study of [3H]ramosetron and [3H]granisetron binding in the cloned human 5-hydroxytryptamine3 receptors. AB - Characteristics of the binding of [3H]ramosetron to cloned human 5 hydroxytryptamine3 (5-HT3) receptors were investigated and directly compared to those of [3H]granisetron binding. Saturation studies revealed that [3H]ramosetron labeled more sites with high affinity (Kd=0.15+/-0.01 nM, Bmax =653 +/- 30 fmol/mg protein) than [3H]granisetron (Kd=1.17+/-0.25 nM, Bmax=427+/-43 fmol/mg protein). Kinetic studies revealed that dissociation of [3H]ramosetron was slower than that of [3H]granisetron. These results suggest that ramosetron is a highly potent 5-HT3-receptor antagonist. PMID- 9869274 TI - A comparative study on the effects of nicotine and GTS-21, a new nicotinic agonist, on the locomotor activity and brain monoamine level. AB - Effects of GTS-21 [3-(2,4-dimethoxybenzylidene)-anabaseine dihydrochloride], a selective nicotinic agonist, on locomotor activity and dopamine turnover were examined and compared to those of nicotine to test if GTS-21 exhibits side effects similar to those of nicotine. GTS-21 had no effect on locomotor activity in mice or dopamine turnover in rats. In contrast, nicotine produced a biphasic effect on locomotor activity. It also enhanced dopamine turnover rates in the striatum and cerebral cortex, suggesting the involvement of dopaminergic systems in the nicotine-induced changes in locomotor activity. GTS-21 exhibits fewer adverse effects, suggesting that it has therapeutic potential for cognitive disorders related to central cholinergic dysfunction. PMID- 9869275 TI - In vitro effects of calcium dobesilate on the responsiveness of spontaneously diabetic rat aorta. AB - We tested the effect of calcium dobesilate (DOBE) in aorta from spontaneously diabetic (BB/wor) rats. The contraction induced by 10(-6) M noradrenaline (NA) in BB/wor rats was smaller than that induced in control rats (1.21+/-0.11 vs 0.82+/ 0.02 g, P<0.01, n=8, respectively) in arteries with intact endothelium. Incubation with DOBE (10(-4) M) impaired the contractions induced by NA in BB/wor rats (1.21+/-0.11 vs 0.67+/-0.01 g, P<0.01, n=8). The effect of DOBE was reversed by 10(-6) M propranolol (0.67+/-0.01 vs 1.20+/-0.60g, P<0.001, n=8, with 10(-4) M DOBE and 10(-4)M DOBE plus 10(-6) M propranolol, respectively). DOBE increased the endothelium-dependent relaxation in arteries from diabetic rats. These findings suggest that DOBE might improve vascular reactivity in BB/wor rats. PMID- 9869276 TI - Possible mechanism of hypoglycemic effect of 4-hydroxybenzoic acid, a constituent of Pandanus odorus root. AB - We studied the hypoglycemic effect of 4-hydroxybenzoic acid, a constituent of the root of Pandanus odorus Ridl. (Pandanaceae, Thai name: Toei-hom), in streptozotocin-diabetic rats. Oral administration of 4-hydroxybenzoic acid caused a decrease in plasma glucose levels dose-dependently in the diabetic rat. The constituent did not affect serum insulin level and liver glycogen content in the diabetic model, but increased glucose consumption in normal and diabetic rat diaphragms. These results suggest that 4-hydroxybenzoic acid produces a hypoglycemic effect mediated by an increase in the peripheral glucose consumption. PMID- 9869277 TI - Distraction from the ground rules? PMID- 9869278 TI - Comparison of different tests assessing neurosensory disturbances after bilateral sagittal split osteotomy. AB - Different methods of assessing functional impairment of the inferior alveolar nerve after sagittal split osteotomy were compared with subjective reports. Thirty patients were tested preoperatively as well as after four days, three weeks, three months, six months, and one year of follow up. All the patients underwent bilateral mandibular ramus sagittal split osteotomy. The sensation of the mandibular teeth was assessed by an electric vitality scanner. Other neurosensory assessments included light touch, two-point discrimination, tactile discrimination, and thermal discrimination. It was shown that electric sensibility-testing of mandibular teeth is a useful method for evaluating neurosensory disturbance after sagittal split osteotomy, and it correlated even better with the patient's subjective evaluation than the other tests used in this study. PMID- 9869279 TI - Soft and hard tissue correction of facial deformity associated with bilateral temporomandibular joint ankylosis. AB - Bilateral temporomandibular joint ankylosis in childhood can result in severe facial deformity characterized by both hard and soft tissue changes. The soft tissue deformity manifests as a submental hump of redundant soft tissue which has been displaced inferiorly due to deficiency of skeletal attachments. Improved results can be produced in these cases if the orthognathic correction is complemented by correction of the soft tissues. The presence of microgenia makes it possible to reach the subplatysmal plane through an intraoral incision allowing for the plication of platysma and relocation of the displaced soft tissues from the cervical to the mental region. PMID- 9869280 TI - Changes of arterial oxygen saturation (SpO2) following push-back operation. AB - This study showed the influence of the push-back operation on the occurrence of sleep-related apnea in cleft-palate patients with an analysis of arterial oxygen saturation (SpO2) during sleep, polygraphic analysis of nasal air flow, and chest wall movements. The postoperative SpO2 was lower than that of the presurgical period in all cases, requiring from five to nine days to recover to presurgical levels. According to polygraphic analysis this depression of SpO2 was caused by peripheral obstructive apnea, while, in spite of the cessation of nasal airflow, chest wall movement continued. PMID- 9869282 TI - Mandibular overdentures supported by two or four endosteal implants. A prospective, comparative study. AB - The aim of this prospective study was to evaluate the effect of the number of implants supporting a mandibular overdenture on the condition of the peri-implant tissues. Sixty edentulous patients (Cawood class V VI) participated in this study. After randomization, thirty patients were treated with an overdenture supported by two IMZ implants (group A) and thirty patients with an overdenture on four IMZ implants (group B). The implants were inserted in the anterior region of the mandible. After three months overdentures were constructed, supported by round bar and clip attachments. A standardized clinical and radiographic evaluation was performed 0, 6 and 12 months after insertion of the denture. One implant was lost (group A) during the healing period. There were no significant differences with regard to any of the studied clinical or radiographic parameters of the peri-implant tissues, neither were significant differences found between the lateral and central implants in group B. None of the patients reported a sensory change in lip or chin region. From this study it is concluded that there seems to be no need to insert more than two endosteal implants to support an overdenture, however, long-term prospective studies are needed to support this notion. PMID- 9869283 TI - Placement of endosteal implants combined with chin bone onlay graft for dental reconstruction in patients with grafted alveolar clefts. AB - Endosteal implants were inserted into grafted alveoli after particulate cancellous bone and marrow grafting in seven patients with cleft lip or palate in conjunction with simultaneous chin bone onlay grafting. In these patients, the alveolar bone height of the bony bridge was insufficient when evaluated by both computed tomographic and periapical radiographic images. The age at first implant surgery ranged from 14 to 28 years. Although four of the seven patients had an uneventful course, three had wound dehiscence, and in all but one of them the exposed chin bone underwent partial or total necrosis. Ultimately all seven implants integrated into the bone, and the alveolar bone height was increased in all but one patient. The results indicate that chin bone onlay grafting with simultaneous implant insertion is useful in patients with cleft lip or palate with insufficient alveolar bone height. PMID- 9869281 TI - Treatment of patients with extreme maxillary atrophy using sinus floor augmentation and implants: preliminary results. AB - Twenty consecutive patients with extreme maxillary atrophy underwent bilateral sinus floor augmentation, either with autogenous bone from the iliac crest or with a combination of autogenous bone and hydroxyapatite. One patient was treated using autogenous bone from the chin region. After a period of three to eight months, three to four implants were placed in each posterior maxilla. Only 10 out of 155 inserted implants were located in the anterior non-augmented maxilla. During the observation period of one to six years, four implants (one of them located in the anterior maxilla) had to be removed prior to prosthetic treatment. Another three implants were lost during the follow up period. This corresponds to a Kaplan-Meier survival probability of 95.4% after 70 months. No statistically significant difference in implant success was observed between women and men (P=0.16). All prosthetic suprastructures are still in function despite these implant losses. Mean peri-implant bone resorption was 1.34 mm with no statistically significant difference between implants placed more mesially and those placed more distally in the augmented area, though a trend could be observed (P=0.058) for a more pronounced bone resorption around implants placed in the premolar region. When a mean mesial and distal bone resorption of >2 mm was considered in the calculation of the success prognosis, the survival probability dropped to 74.7% after 70 months. PMID- 9869284 TI - Alveolar bone growth in response to endosteal implants in two patients with ectodermal dysplasia. AB - The occurrence of alveolar bone growth after placement of endosteal mandibular implants in two edentulous children is reported. The fact that alveolar bone growth occurred in the absence of natural teeth suggests that its growth and preservation is dependent upon biomechanical factors rather than the presence of teeth, as is traditionally thought. PMID- 9869285 TI - Central acinic cell carcinoma of the mandible. Report of a case. AB - Central acinic cell carcinoma is extremely rare; only six cases have been reported in the literature. An unusual case of central acinic cell carcinoma of the mandible in an 84-year-old Japanese woman is presented. This is thought to be the seventh case of central acinic cell carcinoma described in the English literature. PMID- 9869286 TI - Achondroplasia, hypochondroplasia and thanatophoric dysplasia: clinically related skeletal dysplasias that are also related at the molecular level. AB - This is the second of three articles on modern genetic concepts of a number of syndromes and disorders. Three short limb skeletal dysplasias with additional abnormalities of the skull are discussed. All are caused by mutations on fibroblast growth factor receptor 3 (FGFR3). A pathogenetic hypothesis is proposed to explain differences in the severity of short stature, midface deficiency, and craniosynostosis. PMID- 9869287 TI - Recurrent ameloblastoma of the jaws. A follow-up study. AB - Twenty-six cases of recurrent ameloblastoma of the jaws within a 15-year period are presented. They represent 8.9% of all cases of ameloblastoma seen in this period. The highest incidence was found in the third decade of life. There is a higher incidence of recurrences in the mandible (16/26) as compared to the maxilla (6/26). Over 80% of the recurrences presented within five years of primary surgery, emphasising the need for adequate and intensive follow-up during this critical period. Three patients whose lesions were operated on in the mandible showed similar lesions in the maxilla after a period of time. Conservative procedures at primary surgery produced the most recurrences. The cure rate for surgery of maxillary ameloblastoma was comparatively poor. The need for good visibility, adequate access and radical surgery in maxillary ameloblastoma is stressed. PMID- 9869288 TI - Dermoid cyst of the parotid gland. AB - A dermoid cyst occurring within the parotid gland of a 37-year-old Chinese man is reported. The diagnostic difficulties, histopathological features and pathogenesis are considered. PMID- 9869289 TI - Necrotizing sialometaplasia in two patients with bulimia and chronic vomiting. AB - The purpose of this report was to present two unusual cases of necrotizing sialometaplasia of the hard palate, in which the patients were suffering from bulimia and chronic self-induced vomiting. It emphasizes the importance of confirming the diagnosis by incisional biopsy and discusses local mucosal trauma as a possible etiologic factor. PMID- 9869290 TI - Treatment of oral leukoplakia by topical application of 5-aminolevulinic acid. AB - A new therapy for the treatment of oral leukoplakia by 5-aminolevulinic acid (ALA) and photodynamic therapy (PDT) is presented. ALA, a precursor in the biosynthesis of haeme, induces the production of the endogenous photosensitizer protoporphyrin IX which can be used for PDT. Twelve patients, who had been suffering from leukoplakia of the oral mucosa for several years, were treated by ALA-mediated PDT. ALA was used as a topical photosensitizer and 20% ALA cream was applied to the leukoplakia lesion of the oral mucosa for two hours and then light activated at 630 nm, 100 mW/cm2 and 100 J/cm2. Five patients showed complete response to the treatment, four patients showed a partial response and in three patients treatment was unsuccessful. One patient with partial response was retreated, after which the lesion disappeared. PMID- 9869291 TI - Development and histologic characteristics of synovitis induced by trauma in the rat temporomandibular joint. AB - Histopathological changes caused by trauma to the rat temporomandibular joint synovium were examined by light microscopy. The trauma was induced by forced hypermobility of the condyle. Pathological changes of the synovium were found primarily in the anterior pouch of the upper joint compartment. The main findings included surface cell proliferation, increased capillary hyperemia, fibrin deposits on the surfaces of inflamed synovial membranes, and fibrinous adhesion between closely opposed synovial membranes. A gradual change from fibrinous adhesion to fibrous adhesion was also seen. PMID- 9869292 TI - Preliminary study of microsurgical repairs of the inferior alveolar nerve in rats using primary suturing and laser weld techniques. AB - A comparison of primary suturing and a new laser weld technique is described for the microsurgical repair of the inferior alveolar nerve in Wistar rats. A reliable method of exposure of the inferior alveolar nerve has been developed in order to allow intraosseous repairs of the nerve involving suturing with 10:0 nylon and a laser weld technique using an albumin-based solder, containing indocynine cardiogreen, plus an infrared (810 nm wavelength) diode laser. Seven cases of microsuture and laser weld repairs were performed with a 29.4% reduction in total operating time in the laser weld group. Histochemical analysis showed comparable mean neuron counts and mean tracer uptake by neurons for the microsuture and laser weld groups. Giant cell reactions were identified in two of the primary suture cases and axon deflection in three cases demonstrating possible advantages of the laser weld technique which showed no adverse reactions by axons or epineurium to the coagulative repair with the solder. The technique of laser weld repair, on initial analysis, therefore appears comparable with traditional suture repairs and indeed may possess several advantages. Further studies are recommended. PMID- 9869293 TI - Application of combined three-dimensional bone and soft tissue model. A case report. AB - This article describes a new application in three-dimensional modelling. By adding the contours of important soft tissue structures to the bone contours, it is possible to produce a combined bone and soft tissue model. The advantages of this technique are shown in a patient suffering from a parapharyngeal tumour. The model not only enabled the precise localisation of the tumour in relation to the surrounding bone, but also identified important structures like the internal carotid artery in relation to the tumour. PMID- 9869294 TI - Retroperitoneal haematoma and small bowel intramural haematoma caused by warfarin and miconazole interaction. PMID- 9869295 TI - Experience in teaching of cleft lip and palate surgery in Asia. PMID- 9869296 TI - The mammalian Rad24 homologous to yeast Saccharomyces cerevisiae Rad24 and Schizosaccharomyces pombe Rad17 is involved in DNA damage checkpoint. AB - Cell cycle checkpoint proteins play critical roles in maintaining genomic stability and integrity to prevent the development of cancer and hereditary diseases. Here we report the isolation of a novel mouse gene encoding the protein MmRad24 [MmRad24 is the mouse homologue of HRad17, which was described recently by A. E. Parker et al. (J. Biol. Chem., 273: 18340-18346, 1998)], which shares significant sequence and structural homology with the budding yeast Rad24 and its fission yeast counterpart Rad17, both of which are required for DNA damage checkpoints. Confocal microscopy revealed that the green fluorescent protein tagged MmRad24 protein is localized to the nucleus in living cells. Fluorescence activated cell-sorting analysis showed that overexpression of the wild-type MmRad24 in diploid fibroblast WI-38 cells caused a significant G2 arrest of the cell cycle, whereas overexpression of a mutant MmRad24 (mutated on the nucleotide binding site) that likely functions as a dominant-negative protein resulted in a defect in cell cycle arrest after DNA damage treatment as measured by bromodeoxyuridine pulse-chase labeling experiments. Taken together, these results suggest that the mammalian Rad24 protein may function as a critical gatekeeper in DNA damage checkpoint control. PMID- 9869297 TI - The murine Hoxb1 3' RAIDR5 enhancer contains multiple regulatory elements. AB - Homeobox genes play key roles in specifying body part identity during vertebrate embryonic development. Retinoids are signaling molecules involved in the regulation of expression of homeobox genes. We have previously identified an retinoic acid (RA)-inducible enhancer (RAIDR5) located approximately 6.5 kb 3' of the coding region of the murine Hoxb1 gene. This 3' enhancer contains three sequences that are highly conserved in similar RA-inducible enhancers identified in the murine and human Hoxa1 genes and in the chicken Hoxb1 gene. One element, a DR5 RA response element, contributes to the RA inducibility of a Hoxb1 reporter gene construct in F9 cells. In this report, further analysis of the other two elements of the Hoxb1 3' enhancer is reported. The two other sequences, conserved element (CE) 1 and CE2, act as negative elements in cultured F9 cells; when either is mutated, an increase in the beta-galactosidase activity of a Hoxb1 reporter gene construct results. A single Hoxb1 CE2 DNA element:protein binding complex was detected in F9 stem cells, and experiments suggest that this is the same binding protein that recognizes the CE2 element of Hoxa1. In a variant F9 cell line in which both allelic copies of the RA receptor gamma (RARgamma) gene are disrupted, the CE2 binding complex is absent, and this absence correlates with the inability of the CE2 element to function as a repressor of Hoxb1 reporter gene expression in these cells. A single Hoxb1 CE1 binding complex is also detected by gel shift assays in nuclear extracts prepared from both stem and RA-treated F9 cells. This complex contains an Mr approximately 200,000 protein as shown by UV cross-linking. Although the sequences of the CE1 elements of Hoxb1 and Hoxa1 are highly conserved, they differ by two nucleotides. Gel shift analysis shows that either of these nucleotide changes prevents binding of F9 cell protein extracts. When gel shift assays were performed using nuclear extracts prepared from mouse embryos at a time when Hoxb1 mRNA is expressed, i.e., day 9.0, CE1 and CE2 binding complexes identical in mobility to those detected in F9 cells were observed. This suggests roles for both the CE1 and CE2 elements in regulating Hoxb1 gene expression during development. PMID- 9869298 TI - Inhibition of G1 cyclin-dependent kinase activity in cell density-dependent growth arrest in human fibroblasts. AB - The growth of normal fibroblasts in culture ceases as the cells reach saturation density. Although cells in dense cultures express functionally active growth factor receptors, they are essentially refractory to the mitogenic activity of growth factors. Northern blot analysis revealed that immediate early genes, c myc, c-fos and c-jun are induced by mitogen in dense cultures. However, these cells fail to express the late G1 genes as E2F-1, cdc25A, and cyclin A in response to mitogen stimulation. Furthermore, because pRb-phosphorylation is a key event in G1 progression, here we show that in dense cultures, pRb remains in its active (hypophosphorylated) form after stimulation by mitogens. We also show that the kinase activity of cyclin-dependent kinases that are indispensable for the phosphorylation of pRb in late G1 phase was decreased on increasing cell density. The reduced kinase activity may be caused by the observed increase in cyclin-dependent kinase inhibitors and the reduction of cdc25A expression in dense cells. PMID- 9869299 TI - Accumulation of p300 mediates transcriptional repression of simian virus 40 enhancer in undifferentiated F9 embryonal carcinoma cells. AB - The SV40 enhancer is repressed in embryonal carcinoma (EC) cells, and it is also repressed by the adenovirus E1A oncoprotein. Repression by E1A is mediated by the binding of E1A to the p300 transcriptional coactivator. Thus, we examined the role of p300 in the repression of the SV40 enhancer activity in EC cells. We demonstrated that high levels of p300 protein are accumulated in undifferentiated EC cells and that the levels decline dramatically upon differentiation because of the changes of protein stability. Furthermore, we showed that overexpression of p300 does not stimulate the SV40 enhancer activity in undifferentiated F9 cells. However, the activity of a p300 mutant deficient for E1A binding can be restored by the presence of excess E1A. In addition, low-level expression of E1A causes derepression of the enhancer activity in F9 cells. These results indicate that the accumulation of p300 protein participates in repression of the SV40 enhancer activity in undifferentiated F9 cells. PMID- 9869300 TI - The COOH-terminal domain of the focal adhesion kinase induces loss of adhesion and cell death in human tumor cells. AB - Focal adhesion kinase (FAK) is a tyrosine kinase that is linked to signaling pathways between cells and their extracellular matrix. An alternate transcript of the COOH-terminal region of the FAK gene, called FAK-related nonkinase, has been shown to act as an inhibitor of FAK in chicken embryo fibroblasts. We have designed an analogous segment of human FAK, FAK COOH-terminal domain (FAK-CD), and transfected this construct into human tumor cells. Expression of FAK-CD inhibited cell growth in BT474 human breast cancer cells and C8161 human melanoma cells. To characterize the nature of growth inhibition, we developed an inducible system of FAK-CD expression and demonstrated that the induced FAK-CD protein localized to focal adhesions, causing cellular rounding, an irreversible loss of adhesion, and subsequent cell death. In addition, expression of FAK-CD reduced tyrosine phosphorylation of FAK, suggesting that FAK-CD may be a potent inhibitor of FAK in human tumor cells. PMID- 9869301 TI - Activation of the osteopontin promoter by the orphan nuclear receptor estrogen receptor related alpha. AB - Estrogen receptor related alpha (ERRalpha) is an orphan nuclear receptor that is closely related to the estrogen receptor. It is expressed in a variety of adult and embryonic tissues (in particular, at the onset of ossification), as well as in several osteoblastic cell lines. ERRalpha acts as a site-specific, cell specific transcriptional activator. Here, we show that ERRalpha transactivates the promoter of the mouse osteopontin (OPN) gene, the product of which is a marker of the late stages of osteoblastic differentiation. This effect is cell specific and is exerted through derivatives of the ERRalpha response element. Overexpression of ERRalpha in three different osteoblast-like cell lines results in an elevation of the amount of OPN-corresponding mRNA. Therefore, OPN is a target gene for ERRalpha, pointing to the role of the latter in osteoblast differentiation. PMID- 9869303 TI - Psychosocial aspects of stress, health and safety on North Sea installations. AB - In recent years, the North Sea oil and gas industry has been exposed to widespread change, including organizational restructuring, "down-sizing", and increased safety regulation. This article reviews the literature on psychosocial stress, health, and safety among offshore personnel in the light of these developments. Few studies directly compare onshore and offshore populations; the available data suggest that, relative to their onshore counterparts, offshore personnel experience greater anxiety, more sleep problems, and higher work load. Within the offshore population, objective factors (eg, size, age, type of installation, work patterns, and occupational differences), subjective work perceptions, individual differences (age and personality), and health behavior, all play significant roles in relation to health and safety outcomes. However, much of the research in stress and health offshore has methodological limitations and does not allow causal interpretation. The need for prospective studies of the long-term mental and physical health of offshore personnel, and for research in several specific areas, is noted. PMID- 9869302 TI - Stage- and subcellular-specific expression of Id proteins in male germ and Sertoli cells implicates distinctive regulatory roles for Id proteins during meiosis, spermatogenesis, and Sertoli cell function. AB - Immunohistological detection of each of the four Id proteins (Id1-Id4) in sections of mouse testis revealed a unique temporal and spatial expression pattern for each Id protein during spermatogenesis. Furthermore, each Id protein displayed a distinctive, dynamic pattern of subcellular distribution. Id1 was uniquely expressed in MI/MII spermatocytes undergoing meiotic division. Id4 protein was detectable in the cytoplasm of type A1 spermatogonia, as well as in late pachytene and in diplotene spermatocytes. Id2 protein, which was most abundant in Sertoli cell nuclei, was also detectable in pachytene and diplotene spermatocytes, but as with Id4, it was absent from MI/MII cells. In postmeiotic spermatids, Id1, Id2, and Id4 proteins were expressed in a stage- and subcellular specific manner. Expression of Id3 was restricted to Sertoli cell cytoplasm. In malignant seminoma cells, all four Id proteins were abundantly expressed with accompanying changes in their subcellular distribution. The observed expression of Id proteins in postproliferative Sertoli cells and spermatids and during specific stages of meiosis implies novel functional roles for this class of transcriptional regulator during spermatogenesis. PMID- 9869304 TI - Job strain and ambulatory blood pressure among female white-collar workers. AB - OBJECTIVES: The association between job strain and ambulatory blood pressure was studied among female white-collar workers. METHODS: This cross-sectional investigation studied 210 women in high- or low-strain jobs randomly selected from 3183 women of all ages, employed as white-collar workers. The women wore an ambulatory blood pressure monitor for 24 hours during a workday. Mean blood pressures were calculated. Psychological demands and decisional latitude were measured twice (14 months before and 7 days before the blood pressure measurement) with 2 scales recommended by Karasek. RESULTS: Significant differences in blood pressure were found according to current job strain among the women holding a university degree. Their mean blood pressures during work were significantly higher [8.0 mm Hg (1.1 kPa) systolic and 6.4 mm Hg (0.8 kPa) diastolic blood pressure] in the high-strain group than in the low-strain group. Statistically significant elevations in blood pressure over the 24-hour period were also found for women with a university degree. Cumulative exposure to high strain over 14 months was also significantly associated with high systolic blood pressure at work, in the evening, and over a 24-hour period irrespective of other factors related to blood pressure. Among the women without a university degree, the blood pressure differences observed between the job strain groups were less than 1 mm Hg (0.1 kPa) and not statistically significant. CONCLUSIONS: These results provide support for the effect of job strain on ambulatory blood pressure only among female white-collar workers holding a university degree. PMID- 9869305 TI - Job strain and time to pregnancy. AB - OBJECTIVES: The association between fertility and job strain defined as high job demands and low job control has not previously been studied. A follow-up study was conducted with prospective collection of information on job strain among women, achievement of pregnancy, and potential confounding variables. METHODS: A total of 297 Danish couples without previous reproductive experience was followed for a maximum of 6 menstrual cycles from termination of birth control until pregnancy. Job demand and job control were measured by a questionnaire developed by Karasek and his co-workers. RESULTS: The odds ratio and 95% confidence interval (95% CI) for conception per menstrual cycle for women with high job strain was 0.9 (95% CI 0.5-1.5) when compared with that of women in low-strain jobs. Only in secondary analyses restricted to couples with no suspected competitive causes of reduced fertility was a statistically significant reduced odds found for women with high-strain jobs compared with all other jobs. CONCLUSIONS: The main finding of this study did not corroborate a hypothesis of a substantial detrimental effect of job strain on fecundability. PMID- 9869307 TI - Lumbar disc degeneration in relation to occupation. AB - OBJECTIVES: The aim was to study risk factors of lumbar disc degeneration demonstrable with magnetic resonance imaging (MRI) with special emphasis on occupational load and back accidents. METHODS: The subjects in this cross sectional study were 53 machine drivers, 51 construction carpenters, and 60 municipal office workers aged 40-45 years. Data on possible risk factors were available from current structured questionnaires and for 4 and 7 years in retrospect. The prevalence of lumbar disc degeneration L2/L3-L5/S1 was determined with MRI. RESULTS: An increased risk was found for posterior disc bulges among the carpenters and for anterior disc bulges among the machine drivers, but decreased signal intensity was not related to occupation. Car driving was also associated with anterior disc bulges. All signs of disc degeneration were related to a history of back accidents. Disc degeneration was not related to body height, overweight, smoking, or the frequency of physical exercise. CONCLUSIONS: Occupational load affects the risk of disc degeneration of the lumbar spine. Accidental back injuries and motor vehicle driving are associated with an increased risk of disc degeneration. Anterior and posterior disc bulges seem to be related to different types of physical loads. PMID- 9869306 TI - Joint effect of shift work and adverse life-style factors on the risk of coronary heart disease. AB - OBJECTIVES: The joint effect of shift work and certain adverse life-style factors on coronary heart disease (CHD) was studied. METHODS: Base-line measurements were obtained for a 6-year follow-up of an industrially employed cohort (N= 1806), whose shiftwork status was recorded from a questionnaire filled out by a sample of the cohort. The CHD end points (codes 410-414 of the 9th revision of the International Classification of Diseases) were obtained from official Finnish registers. In order that the joint effects of shift work and life-style factors on the risk of CHD could be studied, dichotomized variables and their combinations as a dummy variable system in Cox's proportional hazards models were used. RESULTS: The relative risks were 1, 1.6[95% confidence interval (95% CI) 1.1-2.5], 1.3(95% CI 0.9-2.1), and 2.7(95% CI 1.8-4.1) for the following combinations of shift work (SW) and smoking (SM): SW-&SM-, SW-&SM+, SW+&SM-, and SW+&SM+, respectively; and the corresponding figures for shift work and obesity (BMI > or =28 kg/m2) were 1, 1.2(95% CI0.8-1.9), 1.3(95% CI0.9-1.9), and 2.3(95% CI1.5-3.6), respectively. In both cases the effect was at least multiplicative. For the shift workers the relative risk for CHD rose gradually with increasing numbers of adverse life-style factors, but for the day workers there was no clear dose-response pattern. CONCLUSIONS: Shift work seems to trigger the effect of other, lifestyle-related risk factors of CHD and therefore calls for active prevention among shift workers. PMID- 9869308 TI - The accident process preceding overexertion back injuries in nursing personnel. PROSA study group. AB - OBJECTIVES: This prospective dynamic-population-based study investigated factors involved in the accident process preceding overexertion back injuries among nursing personnel. METHODS: The study covered all reported occupational overexertion back injuries due to accidents among of the approximately 24 500 nurses in the Stockholm County hospitals during 1 year. It was assumed that several factors interact in the accident process. Detailed information was obtained for each injury by interviews with the injured nurse and head nurse. Risks in the physical environment were identified using an ergonomic checklist. RESULTS: During the study 136 overexertion back injuries were reported. Of the 130 nurses participating in the study, 125 had been injured in connection with patient work. Cluster analysis yielded 6 clusters and their pattern of contributing factors. The most frequent injury occurred during patient transfer in the bed or to or from the bed, without the use of transfer devices, when the patient suddenly lost his or her balance or resisted during the transfer and the nurse had to make a sudden movement. However, there were physical conditions, such as shortcomings in the physical work environment or a lack of a transfer device, that compelled the nurses to perform the tasks under unsafe conditions. CONCLUSIONS: The clusters showed a complexity of different kinds of accidents and indicated that the measures for preventing accidents, or for blocking an accident process once started, have to be of different kinds and placed at several different levels in the organization of a workplace. PMID- 9869309 TI - Validity of a self-completed questionnaire measuring the physical demands of work. AB - OBJECTIVES: This study determined the accuracy of workers in quantifying occupational physical demands on a self-administered questionnaire. METHODS: First, a self-administered questionnaire on work postures, manual materials handling, and repetitive upper-limb movements was validated using direct simultaneous observations for 123 randomly selected employees from 6 occupational settings. Second, weight estimation accuracy was assessed on visual analogue scales for 6 manual materials-handling activities using 20 randomly selected employees from 1 occupational setting. RESULTS: At a dichotomous level (ever never), the accuracy of most of the self-reported physical demands was good (sensitivity 60-100%; specificity 56-100%). A more-detailed analysis of the dimensions studied (frequency, duration and amplitude) also showed that the accuracy of the self-reported estimates was satisfactory. Full agreement between the estimated and observed frequency was >60% for most of the manual materials handling activities. In addition the average difference between the estimated and observed duration of the physical demands was found to be small. Finally the average difference between the self-reported and actual weights of various loads was found to be modest. CONCLUSIONS: The self-reported questionnaire used in this study would provide a useful instrument for estimating occupational physical demands and the frequency, duration, and amplitude of these demands in future epidemiologic studies associated with musculoskeletal pain. PMID- 9869310 TI - Cancer mortality of art glass workers in Tuscany, Italy. AB - OBJECTIVES: This study considers the cause-specific mortality from cancer among art glass workers employed in 17 industrial facilities in Tuscany, Italy. METHODS: A cohort of 3390 workers, 3180 men and 210 women, employed at least 1 year, was taken from company payrolls. It was followed between the year each factory started operation, mostly the mid-1950s, and 31 December 1993. The cause specific expected mortality from cancer was computed for men relative to Tuscany rates, specified for gender, 5-year age groups, and calendar year. Separate analyses were carried out for the job title of maker and former and for batch mixers. RESULTS: For 3180 men, the observed mortality was above the expected for larynx [standardized mortality ratio (SMR) 166, 90% confidence interval (90% CI) 90-282], lung (SMR 123, 90% CI 100-151), stomach (SMR 105, 90% CI76-142), and brain (SMR 150, 90%CI 71-282) cancer. Increases for these causes were also found for the makers and formers. Mortality from larynx and lung cancer increased with latency, and significantly increased SMR values were observed for > or =21 years since first exposure. The increasing pattern was also present after adjustment for smoking. CONCLUSIONS: The results showed consistently increased mortality from larynx and lung cancer in the overall cohort and among makers and formers. Stomach and brain cancer was also increased in the overall cohort and among the makers and formers. PMID- 9869311 TI - Anthophyllite exposure and endemic pleural plaques in Kumamoto, Japan. AB - OBJECTIVES: This study explored the high prevalence of pleural plaques in the town of Matsubase in Kumamoto, Japan. METHODS: Small-size chest X-ray film was used for screening, and all persons with pleural plaques were confirmed by computed tomography (CT). The prevalence rate of pleural plaques in the 4 districts of Matsubase and its surrounding towns and cities were also examined. The age-adjusted mortality rate for lung cancer in this town was compared with that of its surrounding towns and cities. RESULTS: Pleural plaques were found in 1357 persons (724 men and 633 women) among the inhabitants who were more than 20 years of age in Matsubase between 1988 and 1993. CT scans ascertained 938 cases with pleural plaques among the 11 14 persons who participated. Thus at least 9.5% of the inhabitants over 20 years of age in this town had pleural plaques. The neighboring towns had a higher rate than the more distant towns. A large-scale open-cast asbestos mine and mill had been in operation in Matsubase between 1883 and 1970. Mineral analysis revealed anthophyllite fibers. Most of the plaques were found in persons who had never worked in the mine or mill. CONCLUSIONS: The high prevalence of pleural plaques in Matsubase was due to anthophyllite exposure, mainly environmental. No mesotheliomas were found, however. These findings agree with those from an earlier study from Finland. PMID- 9869312 TI - Occupational skin and respiratory diseases among hairdressers. AB - OBJECTIVES: The occurrence and causes of hairdressers' occupational skin and respiratory diseases were studied. METHODS: Of a random sample of 500 female hairdressers aged 15-54 years, 355 were available for study. Of the 189 reporting work-related skin and respiratory symptoms in a computer-aided telephone interview on exposure and health, 130 underwent a physical examination, lung function tests, prick and patch testing, and nasal and lung provocation tests. An occupational disease was diagnosed when the causality between exposure and disease was probable and the clinical tests supported the diagnosis. RESULTS: The telephone interview revealed a life-time prevalence of 16.9% for hand dermatoses, 16.9% for allergic rhinitis, and 4.5% for asthma among the hairdressers. In the clinical investigations, the prevalence was 2.8% for occupational dermatoses, 1.7% for occupational rhinitis, and 0.8% for occupational asthma. Ammonium persulfate caused 90% of the respiratory diseases and 27% of the hand dermatoses. Paraphenylenediamine, natural rubber latex, and skin irritation were also causes of hand dermatitis. Allergy to human dandruff (8.6%) and Pityrosporum ovale (12.1%) was common. Previously diagnosed atopic diseases increased the risk for occupational skin or respiratory disease 3-fold (odds ratio 2.9, 95% confidence interval 1.1-7.9). Of the cases, 37.5% (6 of 16 persons) had to change occupations during a 3-year follow-up. CONCLUSIONS: Work-related skin and respiratory symptoms are common among hairdressers. Often a specific cause (eg, ammonium persulfate) can be found if occupational diseases are suspected and diagnosed. Hairdressers with atopic diseases are at risk of developing occupational skin and respiratory diseases. PMID- 9869313 TI - Year of birth and sperm count in 10 Danish occupational studies. AB - OBJECTIVES: Several reports indicate a secular decline of human sperm counts. It is still not known if these findings are artifacts related to shortcomings in the data and applied methodologies. Even less is known about possible mechanisms, but it has been proposed that potential changes may be related to disruption of the hormonal regulation of testicular development in prenatal life. The objective of this study was to examine whether sperm count was related to year of birth. METHODS: An analysis was made of the sperm count of 1196 men participating in 10 cross-sectional occupational sperm studies in 3 regions of Denmark from 1986 through 1995. RESULTS: The median sperm concentration was 63 million per milliliter for men born in 1937-1949 and 52 million per milliliter for men born in 1970 or later, and the median total sperm was 206 million and 117 million, respectively. The inverse relationship between sperm concentration and year of birth was statistically significant even after adjustment for duration of sexual abstinence, season of the year, and study population. However, bias because of differential participation related to age and fertility or lack of comparability across the populations cannot be ruled out. CONCLUSIONS: The apparent decline of sperm count with increasing year of birth is compatible with the hypothesis of a common risk factor for male reproductive health operating in prenatal life or early childhood, but the evidence is circumstantial. Age-related selection bias is an alternative and perhaps not a less likely explanation. PMID- 9869314 TI - Job demands, muscle activity and musculoskeletal symptoms in relation to work with the computer mouse. AB - OBJECTIVES: This study assessed work postures, movements, psychosocial job demands, and shoulder and wrist extensor muscle activity and registered the prevalence of musculoskeletal symptoms of computer-aided design (CAD) operators. METHODS: A questionnaire survey was used to study the use of the computer mouse, psychosocial work factors, and musculoskeletal symptoms among 149 CAD operators. A workplace study was performed using observations, electrogoniometers on the wrists, and electromyography to measure exposures and physiological responses during CAD work among a subgroup of the CAD operators. RESULTS: Musculoskeletal symptoms were far more prevalent for the arm or hand operating the mouse than for the other arm or hand, and women were more affected than men. The symptoms may be related to such risk factors as repetitive movements, static postures (eg, ulnar deviated and extended wrist on the mouse side), and static muscular activation patterns. The risk factors were present due to continuous mouse use and possibly also due to high demands for mental attentiveness, precision, and information processing. CONCLUSIONS: Exposure during work with a computer mouse may present a risk for developing musculoskeletal symptoms. Improvements should focus on introducing more variation. PMID- 9869315 TI - Validity and reliability of self-reported retrospectively collected data on sick leave related to musculoskeletal diseases. AB - OBJECTIVES: The aim was to study the reliability and validity of retrospective data, collected by self-report, on sick leave related to musculoskeletal diseases. METHODS: The study groups consisted of 66 and 306 subjects, for the reliability and validity studies, respectively. They were all part of a wider study of risk factors for musculoskeletal disorders, the REBUS study, conducted in Stockholm in 1993. Reliability was tested using a test-retest design regarding self-reported sick leave related to musculoskeletal diseases in 1970-1993. The validity study comprised the period 1990-1994. Self-reported and registered sick leave data related to musculoskeletal diseases were collected and analyzed regarding concordance. Data about current musculoskeletal disorders and different work-related conditions were collected and analyzed regarding possible effect- and exposure-dependent misclassification. RESULTS: The test-retest reliability study showed the percentage of agreement to be between 0.88 and 0.97, and the kappa values were between 0.73 and 0.93. The validity study of the concordance between the self-reported and registered data showed high agreement and specificity, but the sensitivity was sometimes lower. All the kappa values exceeded 0.50. No effect- or exposure-dependent misclassification was found. CONCLUSIONS: The validity of retrospectively collected self-reported sick-leave data was sufficient for use as a measure of musculoskeletal morbidity in the analyses of associations with work-related conditions. Because of the relatively low sensitivity, such data will underestimate the prevalence of sick leave and should not be used for surveys of morbidity. PMID- 9869316 TI - Suprathreshold intensity and annoyance reactions in experimental challenge to toluene and n-butyl acetate among subjects with long-term solvent exposure. AB - OBJECTIVES: This study explores reactions to low-level chemical challenge, aiming at the development of test procedures for assessing individual sensitivity to smells and chemicals. METHODS: Subjects with symptoms and neuropsychological test results compatible with toxic encephalopathy type 2A (TE-2A) and 2B (TE-2B) and unexposed referents (N=12 in each group) were challenged in an exposure chamber. Toluene exposure was started at 11 mg/m3, and it followed a geometric progression scale with a ratio of 2, until reaching 180 mg/m3. In a counterbalanced design, the subjects were similarly exposed to n-butyl acetate starting at a concentration of 14 mg/m3 and increasing to 228 mg/m3. At each exposure level, smell intensity was measured on a 7-step category scale. Mucous membrane irritation and annoyance reactions were rated on visual analogue scales. RESULTS: Both TE groups showed high sensitivity to the low-level solvent challenge, which provoked immediate annoyance and fatigue reactions. In particular the TE-2B group related smell intensity to various annoyance dimensions during exposure to n butyl acetate, a pattern not observed during toluene exposure. The reference group clearly separated smell intensity and annoyance reactions in both exposure conditions. CONCLUSIONS: The reaction of the TE cases suggests that chemical sensitivity can be distinguished from normal annoyance reactions by the inability to differentiate between smell intensity and an experience of irritation from mucous membranes in air concentrations well below the trigeminal irritation threshold level. Fatigue coreactivity in challenges to single substances below the neurotoxic level may also be important. PMID- 9869317 TI - Audit matrix for evaluating Finnish occupational health units. AB - OBJECTIVES: The aim of this article is to describe how an audit matrix for occupational health units was formulated, what the preliminary results tell about its applicability, and how the matrix can help to implement the principles of "good practice in occupational health services". METHODS: Twelve areas eligible for auditing were selected for the matrix. According to the recently issued principles of good practice, the principles of total quality management and continuous quality improvement, each area was divided into descriptions of 4 quality levels, the lowest being given 0 points and the highest 5-6 points. The maximum total score was 72 points. RESULTS: Of the 30 external audits, no unit had less than 18 points. Nine units had 19-36 points, 17 units had 37-54 points, and 4 units had over 54 points. "Maintenance of work ability" received the highest mean of the given points (4.1), followed by "curative care"(4.0) and "multidisciplinarity"(3.9). The lowest means were for "planning of activities" (3.1), "customer orientation"(3.2), and "quality improvement"(3.2). CONCLUSIONS: The audit matrix proved to be a practical method for auditing occupational health units, revealing both the most common flaws and strengths in the quality of Finnish occupational health services. Because the matrix has been accepted well by occupational health professionals and it manages to classify the units, it is probably a useful instrument for evaluating the performance of occupational health units and promoting good practice. PMID- 9869318 TI - Early diagnosis of cognitive impairment in the elderly with the focus on Alzheimer's disease. AB - In dementia disorders, it can be assumed that the pathological process in the brain has been present for a long time. It is therefore of importance to have a preclinical or an early clinical diagnosis. Obviously, vulnerability genes, such as ApoE-4, can be diagnosed preclinically. As we have no treatment to offer patients with genetic risk factors, genotyping for ApoE-4 is at present of no clinical use. Trained neuropsychologists have today access to sensitive tests which reveal cognitive impairment before the disturbances reach the level of dementia. Laboratory investigations of cerebrospinal fluid have so far yielded no great results. Tau protein appears to be the most sensitive marker, but it is unspecific. Chromogranin A separates early onset from late onset Alzheimer's disease and seems to be a marker for synaptic degeneration. Synaptotagmin was also found to be reduced in patients with early onset Alzheimer's disease. Still we do not know, however, whether these proteins are early markers for degenerative processes in the brain. Laboratory investigations of blood have not yielded markers of use in early or differential diagnosis of dementia disorders. In a study at our own institute, however, we found serum-homocysteine (S-HCY) to be an early and sensitive marker for cognitive impairment. In patients with dysmentia (mild cognitive impairment), no less than 39% had pathological S-HCY levels, indicating insufficient 1-carbon metabolism. PMID- 9869319 TI - Alzheimer-type lesions in Huntington's disease. AB - Cognitive changes in Huntington's disease (HD) are variously related to diffuse cortical atrophy with neuron loss and dystrophic neurites leading to disruption of striato-frontal or limbic circuitries, while recent studies suggest an increasing prevalence of Alzheimer-like lesions in HD brain. A comparative morphological study of 27 autopsy cases of HD (age 34 to 75 years) and of 26 age- and sex-matched non-demented controls was performed. Absence of Alzheimer-type lesions was seen in 33% of HD brains (mean age 49 years); 48% showed early non neuritic tau pathology in limbic areas (Braak stages I and II) without amyloid deposits occurring as early as age 34 years (mean age 54 years), while Braak stages II and III with amyloid plaques were present in 19%, the youngest such HD patient being 42 years (mean age 54 years). In controls, similar tau pathology changes with later onset (age 45 years) and occurrence of amyloid plaques in 26%- all aged over 60 years--were observed. No probable or definite cases of Alzheimer disease (AD) according to CERAD criteria were seen in both cohorts. Those data confirm previous studies on the rare coexistence of HD and AD, although initial stages of Alzheimer-like lesions develop rather early in HD patients, but obviously show less rapid progress even in advanced age. The reasons for the early onset but mild progress of Alzheimer-like lesions in HD and their contribution to cognitive decline await further elucidation. PMID- 9869320 TI - Argyrophilic grain disease: frequency of occurrence in different age categories and neuropathological diagnostic criteria. AB - Argyrophilic grain disease is a progressive degenerative disorder of the human brain which becomes increasingly prevalent with advancing age. The disease entails multiple neuronal systems and results from cytoskeletal degeneration in only a few neuronal types and in oligodendrocytes. Immunoreactions for abnormally phosphorylated tau protein permit identification of the changes. Only a fraction of the emerging abnormal fibrillary material shows a pronounced argyrophilia. Essential for neuropathological diagnosis is assessment of the presence of small spindle-shaped argyrophilic grains in neuronal processes. The anteromedial portion of the temporal lobe bears the brunt of the lesions. Grains generally can be found in abundance in the entorhinal region, the first Ammon's horn sector, the subcortical nuclear complex of the amygdala, and the hypothalamic lateral tuberal nucleus. Frequently, the lesions co-exist with those typically found in Alzheimer's disease or other tauopathies. Owing to the characteristic grains, the disorder easily can be differentiated from other tauopathies. 2661 non-selected brains obtained at autopsy included 125 cases of argyrophilic grain disease (5%) from individuals between 51 and 96 years of age (mean 79 years) . The fact that the same material contained 146 cases of fully developed Alzheimer's disease (6%) supports the view that argyrophilic grain disease is not a rare disorder. Its prevalence with and without concomitant neurofibrillary changes of the Alzheimer type grows with increasing age. Argyrophilic grain disease merits attention because of its frequent occurrence and its potential to cause severe brain dysfunction. PMID- 9869321 TI - Normal and pathological aging--findings of positron-emission-tomography. AB - Normal aging of the brain is predominantly characterized by metabolic changes in the prefrontal cortex. While in middle age there is a trend to hyperfrontality, PET demonstrates in old age a decline of regional cerebral glucose metabolism in frontal areas. In progeric diseases, clinically apparent as premature aging, the metabolic pattern is similar like in normal aging but qualitatively more severe. In patients with the diagnosis of probable Alzheimer's disease (AD) hypometabolism in early dementia is typically present in heteromodal association areas. Hypometabolism then spreads to other cortical and subcortical regions suggesting a characteristic pattern of degeneration that reflects selective vulnerability within limbic-cortical networks. Synaptic plasticity, clinically apparent as cognitive reserve capacity, can be assessed by PET under specific cognitive activation. In AD it is reduced in comparison to age-matched normals and may be influenced by drugs giving trophic support to neurochemical systems. PMID- 9869322 TI - The effects of perturbed energy metabolism on the processing of amyloid precursor protein in PC12 cells. AB - The mismetabolism of amyloid precursor protein (APP), favouring the production of A beta, is considered to be central to the pathogenesis of Alzheimer's disease (AD). However it remains to be established whether the causative factor is the reported toxicity of A beta or reduced production of secretory derivatives of APP which may have trophic or neuroprotective properties. One possible contributory factor to an imbalance in APP metabolism is the impaired cellular energy availability described in AD. The aim of this study was to investigate processing of APP-like proteins following inhibition of oxidative energy metabolism in PC12 cells. Under these conditions, intracellular and secreted APP-like proteins were significantly reduced. Treatment of energy perturbed cells with the lysosomotropic agent chloroquine restored intracellular concentrations of APP like proteins to the control range, while the secretion was completely restored by activation of protein kinase C. These findings raise the possibility that energy related metabolic stress may lead to altered metabolism of APP-like proteins favouring a potentially amyloidogenic pathway. Furthermore, the observation that activation of PKC is able to overcome this potentially pathogenic process has important implications for treatment of AD with the current generation of cholinomimetic drugs, suggesting that such drugs may slow disease progression as well as improve cognitive dysfunction. PMID- 9869323 TI - Abnormalities of mitochondrial enzymes in Alzheimer disease. AB - Abundant evidence, including critical information gathered by Prof. Siegfried Hoyer and his colleagues, indicates that abnormalities of cerebral metabolism are common in neurodegenerative diseases, including Alzheimer's Disease (AD). Alterations in mitochondrial enzymes likely underlie these deficits. Replicable reductions in AD brain occur in the pyruvate dehydrogenase complex (the link of glycolysis to the Kreb's cycle), the alpha-ketoglutarate dehydrogenase complex (KGDHC; the link of Kreb's cycle to glutamate metabolism) and cytochrome oxidase (the link of the Kreb's cycle to oxygen utilization). Available evidence suggests that deficiencies in KGDHC may be genetic in some cases, whereas evidence that the other two enzyme systems have a genetic component is lacking. Additional results indicate that the reductions can also be secondary to other causes including oxidative stress. A variety of data suggest that the mitochondrial insufficiencies contribute significantly to the pathophysiology of AD. PMID- 9869324 TI - The time course of nerve growth factor content in different neuropsychiatric diseases--a unifying hypothesis. AB - Nerve growth factor (NGF) is the prototype of related neurotrophic proteins, the so-called neurotrophins. NGF is essential for proper development of sympathetic and neural crest-derived sensory neurons of the peripheral nervous system (PNS) as well as of the neurons in the cholinergic basal forebrain (CBF). In the mature peripheral and central nervous system (CNS) NGF is also biologically active; NGF facilitates neuronal plasticity and regulates synaptic transmission and connectivity. Besides this well established neurotrophic function, recent findings suggest a role of NGF in neuroimmune and stress-associated processes, which NGF imparts not only as the classical "target-derived messenger", that is retrogradely transported within NGF-sensitive neurons, but also as para- and autocrine cytokine modulating the function of non-neuronal cells. Since neurotrophins are produced in very small amounts in vivo, NGF-sensitive cells have to compete for the limited NGF even under physiological conditions, so that normally only less than 10% of NGF receptors (NGFR) are saturated with their endogenous ligand. Consequently, it is feasable that minute changes in NGF concentrations can influence neuronal function in an extensive way. Hence, one plausible pathomechanism of disease(s) may be that a deficiency in NGF leads to malfunction of NGF-sensitive neurons. The change in NGF concentrations in the course of several diseases, namely during alcoholic and diabetic neuropathy as well as in Alzheimer's disease (AD) and several lesion-models of the CBF, indicates that fluctuations of endogenous NGF concentrations in PNS and CNS follow a distinctive pattern. An initial deficit of NGF at the onset of pathological processes is typically followed by its temporary elevation, during which some neuronal deficits may be partially ameliorated. However, if the disease progresses a decrease of NGF is typically observed, which appears to be a "normalization" of formerly elevated NGF concentrations. In our hypothesis we postulate that after acute or chronic injuries NGF is up-regulated as an intrinsic attempt to regenerate NGF-sensitive neurons. After long-term exposure to noxious processes, however, this compensatory increase of NGF cannot be maintained and eventually breaks down. The extent of such a compensatory up regulation may depend on age and condition of NGF-sensitive neurons as well as on the type of lesion (acute or chronic). Furthermore, we also postulate that an exceeding level of NGF or its chronic elevation could even be detrimental to neuronal functioning under certain conditions. Thus, endogenous NGF has the capacity to modulate and even to compensate different kinds of harmful processes and in this way it may reinstate the homeostatic equilibrium. In our view, it seems to be a more appropriate approach to regard NGF changes as independent of classical constructs of neuropsychiatric diseases. Perhaps our understanding of NGF may even model for a new approach to the aetiology of multifactorial neuropsychiatric disorders. PMID- 9869326 TI - Clinical neurochemistry: developments in dementia research based on brain bank material. AB - Brain tissue obtained at autopsy continues to provide unique opportunities in current dementia research. Not only is tissue analysis still essential for diagnosis, but investigation of neurochemical pathology, at a level of resolution beyond current in vivo imaging, continues to provide new insights into the involvement of neurotransmitter signalling systems. These are relevant to therapy which, with respect to symptoms such as cognitive impairment, psychosis and depression, is currently targeted to specific transmitter (cholinergic, dopaminergic and serotonergic) systems. This paper focuses on dopaminergic, cholinergic and histaminergic parameters in Alzheimer's disease (AD), Dementia with Lewy bodies (DLB) and Parkinson's disease (PD). In the normal striatum the dopamine transporter and D2 receptor exhibit distinct rostral-caudal distributions and D2 binding is affected by genetic polymorphism at the Taq 1A locus. The transporter is reduced in both DLB and PD but not AD, correlating with severity of extrapyramidal dysfunction, and receptor abnormalities are apparent in DLB patients responding adversely to neuroleptics. Striatal nicotine receptors are lost in all 3 disorders, further reduced as a result of neuroleptic medication, and elevated as a result of tobacco use. In the thalamus there are selective reductions in presynaptic cholinergic activity in DLB in the reticular nucleus which relate to symptoms of hallucinations and fluctuating consciousness prevalent in this disorder. In the hippocampus coupling of muscarinic M1 receptors, relevant to response to cholinergic therapy, is impaired in areas most affected by beta-amyloid plaques and intact in less affected areas. Analysis of histamine H2 receptors indicates that, despite presynaptic histamine abnormalities in AD, receptor numbers are normal. Such clinically and therapeutically relevant observations on human brain neurochemistry provide a basis for improving therapeutic strategies and prospects of diagnostic in vivo chemical imaging. PMID- 9869325 TI - Brain-derived neurotrophic factor (BDNF) protects cultured rat cerebellar granule neurons against glucose deprivation-induced apoptosis. AB - In the present study, cell death induced by glucose deprivation in primary cultures of cerebellar granule neurons was examined. Glucose deprivation-induced apoptotic cell death was demonstrated using the terminal transferase-mediated (TdT) deoxyuridine triphosphate (d-UTP)-biotin nick end labeling (TUNEL) method and DNA fragmentation assays. When the effects of different neurotrophins on the survival of cerebellar granule neurons after glucose deprivation were assessed, BDNF, but not NT-3 or NGF, was found to protect cerebellar granule neurons against glucose deprivation-induced cell death. In addition, BDNF treatment increased c-Fos immunoreactivity in the cerebellar granule neurons. These results are consistent with the hypothesis that neuronal death due to glucose deprivation has a significant apoptotic component and that neurotrophins can protect against hypoglycemic damage. PMID- 9869327 TI - C-fos expression in the rat nucleus basalis upon excitotoxic lesion with quisqualic acid: a study in adult and aged animals. AB - A unilateral quisqualic acid lesion was placed in the nucleus basalis magnocellularis of 3- and 24-month-old rats, and the animals were sacrificed at different times post-surgery. The morphology and the number of the cholinergic neurons of the nucleus basalis were analyzed by means of immunohistochemistry for cholineacetyltransferase, in order to evaluate the size and severity of the lesion. Immunohistochemistry for the immediate early gene c-fos was also performed in order to clarify its role in the process of neurodegeneration following the excitotoxin injection. The DNA laddering and TUNEL techniques were used to define the type of cell death involved. At short times (4 hr) the lesion induced alterations in the morphology of cholinergic neurons of the nucleus basalis. Subsequently, a significant decrease in the number of neurons was found in comparison to the contralateral unlesioned side. In the older animals the loss of cholineacetyltransferase immunoreactivity had an earlier onset (4 hr) than in the young (24 hr). C-fos expression was induced by the lesion and not by saline injection in the nucleus basalis and in neighbouring areas of the brain as early as 4 hr after surgery. The c-fos protein was no longer present by 24 hr. Furthermore, the c-fos gene product was consistently absent from the nuclei of cholinergic cells. The aged animals exhibited a slower and smaller increase in c fos as measured by counting the labelled nuclei in the injected area. Analysis of DNA fragmentation did not provide any evidence for apoptosis as the type of cell death involved in the cholinergic degeneration. These results indicate that the c fos protein might have a protective role in the response to excitotoxic lesions. Furthermore, we have shown that the aged brain displays a reduced ability to produce a c-fos-mediated plastic response to the lesion. PMID- 9869328 TI - Neuronal expression of cycline dependent kinase inhibitors of the INK4 family in Alzheimer's disease. AB - Neurodegeneration and cell death in Alzheimer's disease might be associated with aberrant proliferative mechanisms and activation of cell-cycle related events. We reported previously on the elevated expression of the cyclin dependent kinase inhibitor p16INK4a in Alzheimer's disease closely associated with neurofibrillary degeneration. In the present study, we demonstrate that other members of the INK4 family of cyclin dependent kinase inhibitors such as p15INK4b, p18INK4c and p19INK4d that bind directly to cdk4/6 or to complexes of cdk4/6 with D-type cyclins are all elevated. In contrast, no indication of altered expression of the cyclin dependent kinase inhibitors p21Cip1 and p27Kip1 were observed. Inhibitors of the INK4-family were strongly expressed in tangle-bearing neurones and neuritic components of plaques. A much lower expression was also seen in astrocytes. These findings add further evidence to the suggestion that a dysfunction of cell cycle regulation is of critical importance in the pathomechanism of Alzheimer's disease. PMID- 9869329 TI - Acetylcholine in human CSF: methodological considerations and levels in dementia of Alzheimer type. AB - Four different methods to measure acetylcholine (ACh) and choline (Ch) concentration, i.e. thermospray/mass spectroscopy (TS/MS), high pressure liquid chromatography/mass spectroscopy (HPLC/MS), high pressure liquid chromatography/electrochemical detection (HPLC/ECD) and gas chromatography/mass spectroscopy (GC/MS), both latter methods coupled to a solid phase extraction system were compared for their applicability to human lumbar cerebrospinal fluid (CSF). Furthermore, samples from 15 control persons and 11 patients with dementia of Alzheimer-type (DAT) were compared to search for an ACh deficit in the CSF in DAT. GC/MS was the most sensitive, but most laborious method, and HPLC/ECD was acceptably sensitive, reliable and more specific. TS/MS was not specific enough for CSF extracts and HPLC/MS was more specific, but far less sensitive. Thus, only GC/MS and HPLC/ECD were qualified to detect ACh in human CSF extracts. Comparison of GC/MS and HPLC/ECD revealed highly correlated levels of ACh (r = 0.999). Using HPLC/ECD, ACh concentrations were greatly reduced in the DAT group (3.75 +/- 1.40 pmol/ml CSF) as compared to the controls (6.14 +/- 1.39 pmol/ml CSF), but the difference between controls and DAT patients was not statistically significant due to the number of samples below detection limit (8 out of 11 samples in DAT, 7 out of 15 in controls). Ch concentrations were not statistically significant between the two groups. The data show that methodological limitations preclude a widespread clinical application of determining ACh in the human CSF. Despite of reductions of ACh in the CSF in DAT, the determination of Ach in the CSF is not suitable for diagnostic purposes. PMID- 9869330 TI - Role of tryptophan in the elevated serotonin-turnover in hepatic encephalopathy. AB - The increase of the brain levels of 5-hydroxyindoleacetic acid (5-HIAA) in hepatic encephalopathy (HE) suggests an increased turnover of serotonin (5-HT). To study the role of tryptophan on the increased brain 5-HT metabolism in HE, we attempted to monitor brain levels of tryptophan in rats with thioacetamide induced acute liver failure by intravenous infusion of branched-chain amino acids (BCAA). The effect of this treatment on 5-HT synthesis and metabolism was investigated in five brain areas. BCAA-infusions (1 and 2 gm/kg/24 h) increased the ratio BCAA/aromatic amino acids in plasma two- and fourfold, respectively, and lowered both plasma and brain levels of tryptophan. At the higher BCAA-dose all parameters suggesting an altered brain 5-HT metabolism (increased brain levels of 5-HT and 5-HIAA, increased 5-HIAA/5-HT ratio) were almost completely normalized. These results provide further evidence for the role of tryptophan in the elevation of brain 5-HT metabolism and for a potential role of BCAA in the treatment of HE. PMID- 9869331 TI - Sub-regional hippocampal vulnerability in various animal models leading to cognitive dysfunction. AB - Various animal models, involving different brain insults, lead to memory deficits, which can be measured using behavioral tests. In numerous studies, using five different experimental models in rats, we have found that cognitive dysfunction is invariably accompanied by hippocampal CA1 and CA3 pyramidal cells degeneration. However, of these two, the most affected area changes from one model to the other. The present manuscript describes and compares the morphological alterations within the hippocampus in the following experimental models: normal aging, hypoxia, prolonged corticosterone administration, brain ischemia and cholinesterase (ChE) inhibition. In all the above, many hippocampal neurons were severely damaged, however, CA3 pyramidal cells were mostly affected in normal aging and following hypobaric hypoxia, whereas CA1 cells were especially affected following corticosterone administration, global ischemia and ChE inhibition. Several mechanisms, which might be involved in the diverse courses of the lesions are being considered: cerebral oxygen and glucose, glutamate neurotoxicity and calcium involvement. It is anticipated that elucidation of the specific role of CA1 and CA3 hippocampal sub-fields in the various experimental models might help in understanding processes such as age related neuronal degeneration and assist in their prevention. PMID- 9869332 TI - Cytotoxicity of advanced glycation endproducts is mediated by oxidative stress. AB - Non-enzymatic glycation of proteins with reducing sugars and subsequent transition metal catalysed oxidations leads to the formation of protein bound "advanced glycation endproducts" (AGEs). They accumulate on long-lived proteins and are for example structural components of the beta-amyloid plaques in Alzheimer's disease. Since the oxidation of glycated proteins as well as the interaction of AGEs with cell surface receptors produces superoxide radicals, it was tested in BHK 21 hamster fibroblast cells and SH-SY5Y human neuroblastoma cells if AGEs can exert cytotoxic effects on cells. Cell viability was assessed with three independent tests: MTT-assay (activity of the mitochondrial respiratory chain), lactate dehydrogenase assay (release of cytoplasmatic enzymes, membrane integrity) and Neutral Red assay (active uptake of a hydrophilic dye). Two model AGEs, chicken egg albumin-AGE and BSA-AGE, both caused significant cell death in a dose-dependent manner. The cytotoxic effects of AGEs could be attenuated by alpha-ketoglutarate and pyruvate, by antioxidants such as thioctic acid and N-acetylcysteine, and by aminoguanidine, an inhibitor of nitric oxide synthase. This suggests that reactive oxygen species as well as reactive nitrogen species contribute to AGE mediated cytotoxicity. Since AGEs accumulate on beta-amyloid plaques in AD over time, they may additionally contribute to oxidative stress, cell damage, functional loss and even neuronal cell death in the Alzheimer's disease brain. PMID- 9869333 TI - Local transport kinetics of glucose during acute and chronic nicotine infusion in rat brains. AB - Acute and chronic infusion of nicotine is known to result in a distinct increase in local cerebral glucose utilization (LCGU) in several brain structures. The present study addresses the question whether this increase in LCGU is paralleled by a local change in glucose transport in rat brain. Nicotine was infused either acutely for 3 hours or chronically by osmotic minipumps for one week. Local rate constants for glucose transport were measured in brain cryosections using the 3-O [14C]methylglucose method. Local rate constants K1 and k2 were lower in part of the brain structures during acute (-10% to -20%) and in nearly all structures during chronic (-39% to -41%) nicotine. The finding of a decreased glucose transport during chronic nicotine was confirmed by additional experiments of 3-O [14C]methylglucose transfer in an epithelial cell culture. It is concluded that acute and chronic nicotine infusion results in decreased glucose transport although LCGU is either unchanged or increased. PMID- 9869334 TI - Lu 25-109, a combined m1 agonist and m2 antagonist, modulates regulated processing of the amyloid precursor protein of Alzheimer's disease. AB - To examine the effects of the combined muscarinic ml-agonist/m2-antagonist Lu 25 109 on regulated processing of the amyloid protein precursor (APP), we used both transfected cells expressing human muscarinic m1 or m2 acetylcholine receptors, and fresh rat hippocampal slices. Lu 25-109 readily stimulated APPs secretion from HEK 293 cells overexpressing m1, but not m2, receptors, as well as from the hippocampal brain slices. Time-course analyses revealed a rapid (5-35 minutes), and a delayed (55-75 minutes) secretory response to Lu 25-109 with distinct concentration profiles suggesting two distinct cell biological mechanisms. Both responses appeared to reflect post-translational mechanisms because levels of APP message were unchanged after 60 minutes of stimulation with Lu 25-109. In comparison to carbachol, Lu 25-109 had a significantly lower intrinsic activity at muscarinic m1 receptors, compatible with a pharmacological profile as a partial agonist at recombinantly expressed m1 receptors. In as much as stimulation of APPs secretion is associated with reduced formation of A beta peptides, Lu 25-109 may be useful to reduce A beta generation, and thus, slow amyloid plaque formation. Moreover, Lu 25-109 may be useful in promoting the known neurotrophic and neuroprotective biological functions of secreted APPs. PMID- 9869335 TI - Effects of Estradiol (-17beta) on learning, memory and cerebral energy metabolism in male rats after intracerebroventricular administration of streptozotocin. AB - Treatment of adult rats with intracerebroventricularly (i.c.v.) injected streptozotocin (STZ) may provide a relevant animal model of chronic neuronal dysfunction that is characterized by a decrease in both the neuronal metabolism of glucose and the formation of energy. The present study was designed to evaluate whether or not rats treated with triplicate i.c.v. STZ injection would show long-term effects in learning and memory behavior as measured by means of a holeboard test, closed field activity, and passive avoidance behavior over a period of 6 weeks. For this purpose, animals with good performance were discriminated from those with poor performance by the holeboard test before i.c.v. administration of STZ. After a 1-week training period with the holeboard all animals improved their abilities in learning and memory, and the improvement was maintained over the investigation period of 6 weeks in the control group. After i.c.v. STZ working memory (WM), reference memory (RM), as well passive avoidance (PA) behavior decreased, deteriorating progressively during the investigation period. The latter were accompanied by a permanent deficit in cerebral energy metabolism. The ongoing deterioration in behavior and the sustained deficit in cerebral energy metabolism occurring after a triplicate i.c.v. STZ administration lead us to assume that this animal model may be appropriate for the investigation of mechanisms characteristic for sporadic Alzheimer disease. In this context, the effect of Estradiol-17beta (E2) on behavior and energy metabolism was studied. We found that E2 slowed down the i.c.v. STZ-induced deterioration in memory functions, partially compensated the learning deficit, and improved the disturbances in cerebral energy metabolism to the extent that it was almost completely normal again. These findings underscore the beneficial effect of E2 in dementia disorders. PMID- 9869336 TI - Effect of propentofylline on hippocampal brain energy state and amyloid precursor protein concentration in a rat model of cerebral hypoperfusion. AB - An animal model of graded cerebral hypoperfusion achieved by way of 2- and 4 brain vessel occlusion (vo) was used to test the effect of chronic propentofylline (PPF) administration on hippocampal energy state and amyloid precursor protein (APP) concentration. For this purpose, forty adult rats were subjected to stepwise and permanent 2 vo and 4 vo and PPF in a dose of 25 mg/day per kilogram body weight was continuously administered intraperitoneally for 1 week or 3 weeks, respectively. During the final steady-state experiment arterial blood parameters and blood gases (mean arterial blood pressure, PO2, pCO2, pH, hematocrit, hemoglobin, body temperature) were measured. Brain tissue concentrations of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine were measured in rat hippocampus by high-pressure liquid chromatography (HPLC) analysis; APP concentration was determined using Western blot techniques. In adult rat brain, long-term PPF treatment induced a striking increase in hippocampal ATP concentration which was paralleled by an enhancement of the ATP/ADP-ratio. Specifically, after 2 vo with 1 week of PPF treatment the ATP concentration in hippocampus was enhanced by 18% when compared with 2 vo without PPF treatment, whereas after 3 weeks PPF administration with 4 vo the concentration of ATP was enhanced by 31%. Furthermore, PPF reduced the tissue concentration of adenosine from 17.75 +/- 2.79 to 8.83 +/- 3.68 pmol/mg wet weight during 4 vo if compared with animals without PPF. In addition, the APP concentration was enhanced by 35% after 1 week PPF administration when compared with non PPF treated animals. In summary, the results demonstrated that chronic administration of PPF induced an enhancement in ATP concentration in adult rat hippocampus under conditions of long-term brain vessel occlusion. Interestingly, 1 week PPF seems to have a stimulating effect on APP which may act neuroprotectively. Thus, PPF may protect hippocampal neurons from chronic ischemic damage. In conclusion, PPF may have some beneficial effects and could be used in the treatment of patients with chronic cerebrovascular disorders or neurodegenerative diseases such as vascular dementia. PMID- 9869337 TI - Automated static perimetry as a screening method for evaluation of retinal perfusion in diabetic retinopathy. AB - The aim of our study was firstly to assess whether areas of capillary non perfusion in diabetic retinopathy are associated with reduction of retinal light sensitivity and secondly to assess whether automated perimetry can serve as a screening method for evaluation of retinal perfusion. 32 eyes with diabetic retinopathy and 30 eyes of controls underwent visual field testing on the 30-2 program of the Humphrey field analyser and fluorescein angiography. The results are compared with the 'superimposition technique'. In 93.4% of our diabetic eyes (30 from 32) areas of capillary non-perfusion demonstrated by fluorescein angiography were associated with areas of reduced retinal sensitivity. The correlation between visual field defects and areas of reduced retinal perfusion was significant with probability more than 99% (P<0.01). In view of our findings we recommend automated static perimetry as a very sensitive method for evaluation of retinal perfusion in diabetic patients. PMID- 9869338 TI - The use of the G1 and Octosmart programs in detecting temporal changes in the visual field. AB - PURPOSE: The Octopus program Octosmart is able to classify visual fields into six classes. In the program a horizontal bar indicates these classes, and an indicator points to the most probable position, related to the measured pathology. The width of this dashed indicator shows the range of possible fluctuations in the measurement and, therefore, its precision. This study sets out to analyse the suitability of this display mode using other visual-field index data. METHODS: The visual fields of 83 glaucomatous eyes of 61 patients of various etiological groups and glaucoma suspects were studied for periods varying from 1 to 5 years in a retrospective study. All examinations were performed with the G1 Octopus program and analyzed with the Octosmart program. The statistical significance of linear trends of the visual-field indices, mean defect (MD) and corrected loss variance (CLV), and the class shown by the indicator (POI = position of indicator) were determined, and their regression coefficients were analyzed by means of a linear trend test as a function of time. RESULTS: Of the sample of 83 tested eyes, a total of 18 significant trends were recorded after five examinations. All visual-field indices showed a trend towards amelioration. CONCLUSIONS: The 18 significant trends observed must be attributed to perturbing long-term fluctuations and, despite their statistical significance, are of little clinical value. It is questionable whether an increased number of examinations per eye would have attenuated the threshold fluctuations sufficiently to make the change in field class more reliable. PMID- 9869340 TI - Acute onset transient ectropion associated with corneal ulcer. AB - We report a case of transient ectropion associated with corneal ulcer. The corneal ulcer responded well to treatment with fortified gentamicin and cefazolin. As the corneal ulcer healed, the eyelid simultaneously returned to its original position. We suggest that in the presence of involutional eyelid changes, even a minimal additional factor, such as conjunctival edema and blepharospasm, may cause transient ectropion. PMID- 9869339 TI - Retinal microvasculopathy in human immunodeficiency type 1 (HIV)-infected children. AB - PURPOSE: To report two cases of retinal peripheral lesions in vertically HIV infected children. METHODS: Complete ophthalmological examinations were performed in both cases over a period of four years. RESULTS: We found retinal peripheral lesions in four eyes of two patients who were referred to us for routine examination. Both cases showed focal, segmental and white patches around the vessel wall in the far periphery and at the equator. These findings hardly changed after 3.5 and 2.5 years of follow-up, respectively. We suppose the cause of these lesions is an immune-mediated reaction. CONCLUSIONS: Even although there is a low incidence of retinal alterations in pediatric patients infected by HIV, we believe that complete and systematic ophthalmic examinations should be performed in these children. PMID- 9869341 TI - Ocular hypotony after laser suture lysis following trabeculectomy with mitomycin C. AB - BACKGROUND: To report our experience with laser suture lysis (LSL) following trabeculectomy with mitomycin C, its timing, effectiveness and related complications. METHODS: We retrospectively examined 38 consecutive eyes of 36 patients that underwent LSL following trabeculectomy with mitomycin C. RESULTS: The mean preLSL intraocular pressure (IOP) was 27.0 mm Hg (SD 6.3, range 16-39 mm Hg) and the postLSL IOP (IOP 1 h after the last session of LSL) was 16.0 mm Hg (SD 7.2, range 3-31 mm Hg). Following the LSL in 7 eyes (7 of 38) hypotony (IOP<6 mm Hg, lasting more than 24 h) developed. Two groups were defined. In group I no hypotony was found after LSL and group II went through a period of hypotony. The time interval between surgery and LSL was significantly shorter in group II (mean 5.7, SD 7.5, range 1-19 days), compared to group I ( mean 14.7, SD 13.0, range 1 44 days) (p=0.041). The mean final IOP (IOP measured at the last visit) was 13.3 mm Hg (SD 3.4, range 6-20 mm Hg) and mean follow-up was 6.1 months (SD 3.9, range 2.0-15.2 months). No hypotony was found at final examination. PostLSL IOP and final IOP were significantly lower in group II (p=0.002 and p=0.024 respectively). IOP reduction by LSL was significantly greater in group II (p=0.046). CONCLUSION: LSL is an effective and safe procedure to lower the IOP following trabeculectomy with mitomycin C. Early application of LSL results in lower final IOPs, but has a higher risk of hypotony. PMID- 9869342 TI - Outcome of late probing for congenital nasolacrimal duct obstruction in Singapore children. AB - OBJECTIVE: To evaluate the outcome of late probing in a mixed Asian children population with congenital nasolacrimal duct obstruction (CNLDO) and whether probing was associated with an increased risk of infection. METHOD: A retrospective clinical study was conducted. A clinical diagnosis of CNLDO was defined in an infant who presented with a history of tearing and/or eye discharge up to 1 year of age with no other accompanying ocular pathology. All patients with a presumed diagnosis of CNLDO who had probing after 12 months of age were included in our study. Prior to probing, most patients had a trial of conservative treatment with massage and/or topical antibiotics. Intra-operative patency of probing was determined when metal to metal contact of the probe and forceps was achieved and/or when fluorescein dye was recovered from the nose after syringing. Successful probing was defined as a resolution of symptoms within 1 month after probing. RESULTS: Seventeen patients involving 19 eyes fulfilled our inclusion criteria for this study. Fourteen (82%) infants were Chinese, two Malays (12%) and the remaining one Indian (6%). There were 10 males (59%) and 7 females (41%) and the mean age at probing was 2.2 years (range, 14 months to 5 years 6 months). The subjects were followed-up post-operatively for a mean duration of 21.2 months (range, 2 months to 8 years). Successful probing in our series was 89.5% (17/19 eyes). Of the two eyes with persistent tearing, one resolved with massage about 9 months after the unsuccessful probing; the other was referred to the oculoplastic service for further management. CONCLUSION: Late probing after 13 months old appears to be an effective approach in the management of CNLDO in Asian infants. PMID- 9869343 TI - Pterygium excision with free conjunctival autograft (FCG) versus postoperative strontium 90 (90Sr) beta-irradiation. A prospective study. AB - In a prospective study the results of a pterygium excision in 54 patients (57 eyes) who underwent a superficial free conjunctival autograft (FCG) were compared to those of patients who were treated with postoperative 90Sr-irradiation. In 51 cases the minimum follow-up was six months, the maximum follow-up seven years. We divided the study up into a randomized part and an open part. In the randomized part, surgery of a primary pterygium was performed in 25 eyes, of which 16 were treated with a FCG and compared with 9 eyes with primary pterygium surgery and postoperative 90Sr beta-irradiation. In the same period 16 eyes were treated because of a recurrent pterygium: 8 with FCG and 8 with 90Sr-irradiation. In the open part of the study 16 eyes with primary pterygium were successively treated with FCG alone. The results showed in the randomized, as well as in the open study on primary surgery with a minimum follow-up of six month, one recurrence in each of the FCG-groups (2 out of 31 eyes = 6.4%), and no recurrences in the 90Sr group (0%). In the randomized group of patients treated for a recurrent pterygium one recurrence developed in the FCG group (1 out of 8 eyes = 12.5%) and one in the 90Sr-group (1 out of 7 = 14.6%). Analysis of other clinical parameters showed that postoperative treatment with corticosteroids, nonsteroidal anti-inflammatory drugs, and artificial tears was necessary for a longer period in the FCG group than in the 90Sr-group. PMID- 9869344 TI - Fibrinolytic activity in retinal vein occlusion. AB - PURPOSE: Impaired fibrinolytic function is a common finding in patients with thrombotic disease. The present study was initiated to evaluate the fibrinolytic response to a venous occlusion test (VOT) in patients with retinal vein occlusion. METHODS: Euglobulin clot lysis time (ECLT), tissue plasminogen activator (t-PA) activity, and plasminogen activator inhibitor (PAI) activity were measured before and after VOT in a group of 26 consecutive patients presenting with retinal vein occlusion and in 15 healthy age- and sex-matched controls. RESULTS: Before VOT (baseline), a higher proportion of patients (54%) had an ECLT of more than 5 h compared with controls (6.7%) (p = 0.0027) indicating decreased overall fibrinolytic activity. Patients had non significantly lower t-PA and higher PAI activities compared with controls. After VOT, a higher proportion of patients (34.6%) had an ECLT of more than 5 h compared with controls (6.7%) (p = 0.05). Patients had significantly lower t-PA activity (p = 0.0232) and significantly higher PAI activity (p = 0.0292). Subgroup analysis revealed that patients with an ECLT of more than 5 h had significantly higher levels of PAI activity at baseline (p = 0.0326) and after VOT (p = 0.0184) compared with patients with an ECLT of less than 5 h. However, t PA activity was significantly higher (p = 0.0153) at baseline, and non significantly higher after VOT in patients with an ECLT of more than 5 h when compared with patients with an ECLT of less than 5 h. CONCLUSIONS: These findings suggest that impaired fibrinolysis due to increased PAI activity may play a role in the pathogenesis of retinal vein occlusion. PMID- 9869346 TI - Isolated lesion of the medial orbital wall following endonasal surgery. Isolated fractures of the medial orbital wall. AB - BACKGROUND: Isolated fractures of the medial orbital wall are infrequent. The diagnostic triad includes: adduction block, exotropia with diplopia in all directions of gaze, positive passive duction in abduction. Sometimes a slight enophthalmos is present. Computed tomography shows the extension and the seat of the fracture. CASE REPORT: The authors illustrate the case of a 60 year old male who presented with a breach of the medial orbital wall following endonasal surgery. RESULTS: The patient was successfully operated using an iliac bone graft inserted via an eyebrow-nasal cutaneous approach, after a previous attempt with a transconjunctival approach performed in another hospital had failed. A good functional and aesthetic result was observed within the first year after surgery. After almost 11 years a full adduction is still present and diplopia is absent. CONCLUSION: The authors underline the importance of an early diagnosis and prompt surgical treatment. The fat-muscle entrapment should be removed and the bone defect closed. A close cooperation between ophthalmologist and plastic surgeon is suggested. PMID- 9869345 TI - Primary use of silicone oil tamponade in the management of perforating globe injury secondary to inadvertent local anaesthesia injection for ophthalmic surgery. AB - Perforating and penetrating globe injuries secondary to peribulbar and retrobulbar anaesthesia are often complicated by vitreous haemorrhage and retinal detachment. We describe the effectiveness of primary silicone oil tamponade in the repair of three perforated globes secondary to local anaesthesia for ophthalmic surgery. Three patients with axial myopia had peribulbar and retrobulbar anaesthesia for extracapsular cataract extraction (two patients) and cryotherapy (one patient). All eyes sustained a vitreous haemorrhage obscuring the view to the fundus. Retinal detachments were detected by B-scan ultrasound. In all eyes, scleral buckling, pars plana vitrectomy and silicone oil tamponade were performed as a primary surgical procedure. All the patients had complete anatomic reposition. In two patients, after two years follow-up, visual acuity was between 6/12 to 6/36 with the retina attached and no proliferative vitreoretinopathy (PVR). The third patient had blind painful eye and enucleation was performed. Primary use of silicone oil tamponade, in the management of perforated globe with retinal detachment due to local anaesthesia injection, is recommended. PMID- 9869347 TI - Treatment of deep and shallow intrabony defects. A multicenter randomized controlled clinical trial. AB - This prospective multicenter intra-individual randomized controlled clinical trial was designed to compare the efficacy of guided tissue regeneration (GTR) with bioresorbable barrier membranes versus access flap surgery, in intrabony defects. 2 similar defects were selected in each of 23 patients and randomly assigned to 1 of the 2 treatments. Surgery consisted of an identical procedure except for the omission of the barrier membrane in the flap control sites. At 1 year, probing pocket depth reductions were 4.3+/-2.3 mm in GTR treated sites and 3.0+/-1.5 mm in the flap control sites (p=0.02, paired t-test). Clinical attachment level (CAL) gains were 3.0+/-1.7 mm in the GTR sites and 1.6+/-1.8 mm in the control sites (p=0.009, paired t-test). A subset analysis, performed according to the initial depth of the intrabony component of the defects (INFRA), indicated that in shallow defects (INFRA < or =3 mm) treated with the access flap alone, CAL gains were 1+/-1.5 mm, while in deep ones (INFRA > or =4 mm) they were consistently greater (1.9+/-1.9 mm). The % CAL gains, calculated as the % of the baseline intrabony component depth, however, were almost identical in the 2 subpopulations (45.8+/-64.7% in shallow and 43.8+/-37.6% in deep defects). Similarly, in the GTR sites, linear CAL gains were greater in deep (3.7+/-1.7 mm) than in shallow defects (2.2+/-1.3 mm), but no differences were observed in terms of % CAL gains (76.7+/-27.7% and 75.8+/-45%, respectively). The frequency distribution of CAL changes expressed as %s of the baseline INFRA indicates that most of the sites treated with GTR (73% in shallow and 92% in deep defects) gained 50% or more CAL. Furthermore, many defects (64% of shallow and 33% of deep defects) reached 100% of CAL gain. The present study demonstrated that: (i) GTR with bioresorbable barrier membranes resulted in a significant added benefit in comparison with access flap alone; (ii) the linear amounts of CAL gains were greater in deep than in shallow defects; (iii) CAL gains expressed as %s of the baseline depths of the intrabony component, were similar in shallow and deep defects; (iii) the regenerative procedure tested in the present study resulted in CAL gains equal to the depth of the intrabony component of the defect in some, but not in most of the instances. PMID- 9869348 TI - Sonic and mechanical toothbrushes. An in vitro study showing altered microbial surface structures but lack of effect on viability. AB - The purpose of the present study was to compare the in vitro effects of a mechanical and a sonic toothbrush on the viability of Actinomyces viscosus, the rationale being that induction of irreparable microbial damage resulting from aggressive mechanical action or sonic energy, may inhibit or disrupt the process of successional colonization. Cultures of A. viscosus were grown to a standardized optical density and subdivided into 3 treatment groups of 20 specimens each. Treatment groups consisted of an untreated control and exposure to a mechanical or sonic toothbrush for 15, 30, 45, and 60 s. Subsequent to the prescribed treatment, samples were taken from each specimen dish, subcultured, and the number of CFUs determined. Additional samples were obtained for negative staining and examination by electron microscopy. The mean number of CFUs for each treatment group at each treatment interval were statistically analyzed by ANOVA and multiple pairwise comparisons. Results showed a significant main effect for toothbrushes (p<0.0001) and exposure time (p<0.01), but only marginal significance for the interaction of toothbrush with exposure time (p<0.055). Post hoc tests showed a significantly greater number of CFUs for the sonic toothbrush compared to both the untreated control and mechanical toothbrush groups. Electron microscopic examination revealed a decrease in aggregation tendency and loss of fimbriae in the sonic toothbrush group. Based on the lack of morphologic evidence that would indicate cell damage and the increase in CFUs over that of the control group, it appeared that neither the mechanical or sonic toothbrushes affected cell viability. PMID- 9869349 TI - Analysis of serum antibody responses to periodontopathogens in early-onset periodontitis patients from different geographical locations. AB - Serum antibody specificity to oral micro-organisms was used to delineate the pathogens associated with early-onset periodontal diseases in a Turkish population. Additionally, comparison of the findings to those derived from a clinically similar US patient population described differences in bacterial specific antibody between these 2 geographic regions. Serum from 89 (LJP), 86 (RPP) and 94 (normal) subjects was analyzed (ELISA) to determine IgG antibody to 14 oral micro-organisms. All LJP patients from Turkey exhibited elevated antibody levels to A. actinomycetemcomitans (serotypes c and a significantly increased), while antibody levels to A. actinomycetemcomitans Y4 and JP2 (serotype b) were significantly higher in US LJP patients. 50% of the Turkish RPP patients also showed elevated anti-A. actinomycetemcomitans antibody, although the US RPP patients exhibited significantly higher antibody levels and frequency of elevated antibody to the A. actinomycetemcomitans serotypes. Healthy subjects and LJP and RPP patients from the US exhibited higher antibody levels to all 3 P. gingivalis serogroups compared to those from Turkey, although, the frequency of elevated antibody to the P. gingivalis serogroups was significantly higher in LJP and RPP patients from Turkey than from the US. Interestingly, 87% and 77% of the LJP patients in the Turkish population had elevated antibody responses to P. gingivalis and E. corrodens, respectively, which was not observed in the US LJP patients. These data suggested that considerable variation exists in the systemic antibody levels to periodontopathogens between these 2 countries. This supports potential differences in subgingival colonization or antigenic composition of these pathogens between patient populations from different geographical regions. PMID- 9869350 TI - Crevicular fluid prostaglandin E2 levels in periodontitis-resistant and periodontitis-susceptible adults. AB - The aim of this cross-sectional study was to determine concentrations of prostaglandin E2 (PGE2) in gingival crevicular fluid (GCF) in a cohort of periodontal disease-resistant (PDR) adults with chronic gingivitis but with minimal evidence of bone loss, and to compare these data with GCF-PGE2 levels in patients with untreated chronic adult periodontal disease (CAPD). 20 PDR and 35 CAPD subjects with mean (+/-se) ages 52.4 (+/-2.9) and 43.7 (+/-1.2) years respectively, were recruited. GCF was sampled from 6 sites in each PDR subject and 4 sites in each CAPD subject. The GCF-PGE2 concentrations were determined by enzyme immunoassay (Assay Designs). Whole mouth medians of site-specific GCF-PGE2 concentrations were calculated for each subject. The means of the median GCF-PGE2 concentrations were: PDR 54.94+/-4.06 ng/ml; CAPD 41.57+/-2.91 ng/ml (p=0.009). We hypothesise that the higher concentrations of PGE2 in the PDR group may be associated with the proliferating pocket epithelia of the chronic gingivitis. In the CAPD cohort, there were no differences in GCF-PGE2 concentrations between subgroups of smokers (n=13), ex-smokers (n=11) and non smokers (n=11). In the PDR cohort, 19/20 subjects were non-smokers. PMID- 9869351 TI - Changes in the prevalence of residual pockets and tooth loss in treated periodontal patients during a supportive maintenance care program. AB - Bleeding on probing and the presence of deep periodontal pockets are considered to be the best site-specific indicators for periodontal disease progression during the maintenance phase of periodontal therapy. A major emphasis of supportive periodontal care (SPC) programs, therefore, has been the control of bleeding pockets. This investigation retrospectively evaluated the changes in the prevalence of bleeding on probing, periodontal pockets, bleeding periodontal pockets and the prevalence of tooth loss in a random sample of 273 periodontal patients participating in a supportive maintenance care program at a University Clinic. During an observation period of 67+/-46 months (range 5 months to 23 years), the overall incidence of all causes of tooth mortality was 0.23+/-0.49 teeth per patient per year of observation. 56% of subjects, however, did not experience any tooth loss, while less than 10% of patients lost more than 3 teeth. Thus, participation in the SPC program was effective in preventing tooth loss in the majority of patients. During the SPC period, however, a significant increase in the prevalence of periodontal pockets, and of bleeding on probing positive periodontal pockets, in particular, was observed. At completion of active periodontal therapy, 56.4% of patients were free from bleeding pockets. This decreased to a mere 13.6% at the latest SPC evaluation. The observed increases in the number of bleeding pockets was significantly associated with: longer times since completion of active periodontal therapy, more advanced periodontal diagnosis, higher %s of bleeding sites in the dentition, cigarette smoking, lack of inclusion of periodontal surgery in the active treatment phase, tooth loss, and the response to the active phase of periodontal treatment. The data presented in the paper indicate that the observed increase in the prevalence of bleeding pockets and tooth loss was not homogeneously distributed in the studied SPC population. Rather, high risk groups of individuals could be identified. It is suggested that better knowledge of risk indicators may lead to improved and more efficient risk management efforts during periodontal maintenance care. PMID- 9869352 TI - An in vivo study of the chlorhexidine release profile of the PerioChip in the gingival crevicular fluid, plasma and urine. AB - The release profile of chlorhexidine from the PerioChip (Chip), a biodegradable local delivery system that contains 2.5 mg of chlorhexidine gluconate (CHX) in a cross-linked hydrolyzed gelatin matrix, into the gingival crevice, was evaluated in an in vivo, open label, single-center, 10-day pharmacokinetic study conducted on 19 volunteers with chronic adult periodontitis. Each volunteer had a single chip inserted into each of 4 selected pockets, with probing pocket depths of between 5-8 mm, at time 0. Gingival crevicular fluid (GCF) samples were collected using filter paper strips prior to Chip placement and at 2 h, 4 h, 24 h and 2, 3, 4, 5, 6, 8, and 9 days post-Chip placement. The GCF volume was measured using a calibrated Periotron 6000. Blood samples were collected at times 0, 1, 4, 8, 12 h and 5 days post-dosing. Urine was collected as a total 24-h specimen immediately post-dosing and 2 single samples at time 0, prior to dosing, and 5 days. The CHX was eluted from the paper strips and the CHX levels in GCF, blood and urine quantified using HPLC. The results indicate an initial peak concentration of CHX in the GCF at 2 h post-Chip insertion (2007 microg/ml) with slightly lower concentrations of between 1300-1900 microg/ml being maintained over the next 96 h. The CHX concentration then progressively decreased until study conclusion with significant CHX concentrations (mean=57 microg/ml) still being detectable at study termination. CHX was not detectable in any of the plasma or urine samples at any time point during the study. These results indicate that the PerioChip can maintain clinically effective levels of CHX in the GCF of periodontal pockets for over 1 week with no detectable systemic absorption. PMID- 9869353 TI - Risk of severe periodontal disease in a Swedish adult population. A cross sectional study. AB - In this study, potential risk factors for severe periodontal disease were identified in a cross-sectional sample from the county of Jonkoping, Sweden. 547 adults 20-70 years of age were categorised clinically and radiographically by level of periodontal disease experience. These levels were used to divide the sample into groups--individuals without any reduction in periodontal bone level (60%) and those with severe periodontal bone loss (13%)--which were then used in univariate and multivariate logistic regression analyses as dependent variable. Demographic, socio-economic, general health, smoking habits, clinical, and dental care variables were used in the different regression analyses. In the univariate model, age (20-70 years) was found to be correlated with more severe periodontal disease experience (odds ratio: 1.13; 95% CI: 1.10-1.17). The association with periodontal disease was more pronounced for the older age groups (50, 60, and 70 years). A negative financial situation was also related to severe periodontal bone loss when regressed univariately (odds ratio 2.20 [95%: 1.04-4.68]). Moderate-heavy smoking (> or =10 cigarettes/day) appeared to be associated with severe periodontal destruction with an odds ratio of 9.78 (95% CI: 3.62-36.42). Of the clinical variables in the univariate model, higher mean levels of supragingival dental plaque and the presence of subgingival calculus were related to more severe periodontal disease with odds ratios of 1.02 (95%: 1.01-1.03) and 2.96 (95%: 1.50-5.88), respectively. When the same variables were regressed multivariately, age (continuous) (odds ratio 1.17 [95% CI: 1.12-1.22]), moderate heavy smoking (odds ratio 11.84 [95% CI: 4.19-33.50]), and higher mean levels of plaque (odds ratio 1.02 [95% CI: 1.00-1.03]) remained significant. Light smoking (1-9 cigarettes/day) was not significantly associated with severe periodontal disease in the 2 regression models. The present study demonstrated that smoking, greater age, and higher mean levels of plaque are potential risk factors for severe periodontal disease in this specific population. PMID- 9869354 TI - Gingival fluid IL-1beta in postmenopausal females on supportive periodontal therapy. A longitudinal 2-year study. AB - Posterior interproximal alveolar bone in 59 women, within 5 years after menopause, was assessed at baseline and after 2 years of supportive periodontal therapy (history of moderate/advanced periodontitis) using digitized image analysis. Baseline lumbar spine bone mineral density, smoking status, and yearly serum estradiol (E2) levels also were obtained to group subjects. An additional 16 non-periodontitis postmenopausal women were followed 2 years for clinical and estrogen status. 2-min GCF IL-1beta levels averaged from 2 baseline periodontal pockets (in periodontitis subjects) and 2 non-periodontitis sites (in non periodontitis and periodontitis subjects) were determined with an enzyme immunoassay. A progressive and stable site were also monitored every 6 months for GCF IL-1beta in 15 patients. Results after 2 years indicated that 17 subjects had no posterior interproximal sites losing > or =0.4 mm of alveolar crest bone height, while 13 subjects had > or =3 such sites. Using analysis of variance, none of the above clinical groupings resulted in a significant difference in mean baseline or longitudinal GCF IL-1beta levels. However, when subjects who lost alveolar crest bone height were considered, E2-sufficient subjects had significantly depressed baseline GCF IL-1beta (in past-periodontitis sites) compared to E2-deficient patients (9.1+/-2.1 versus 31.7+/-10.2 pg/2-min sample, p<0.05), suggesting E2 influences gingival IL-1beta production in progressive periodontitis patients. PMID- 9869355 TI - A 5-year follow-up of 16 patients treated with coralline calcium carbonate (BIOCORAL) bone replacement grafts in infrabony defects. AB - A resorbable coralline calcium carbonate graft material (BIOCORAL) (CalCarb) was evaluated as a bone replacement graft in human periodontal osseous defects. Following initial preparation and re-evaluation, flap surgery was carried out. Bone defects were curetted and root surfaces subjected to mechanical debridement and conditioning with tetracycline paste. The bone defects were grafted with CalCarb, and the host flaps replaced or slightly coronally positioned. Weekly, then monthly deplaquing was performed until surgical reentry at 6-12 months. Patients were then followed on approximate 3 month recalls for > or =5 years. Significant clinical changes included improvement in mean vertical clinical probing attachment level from 5.7 mm at surgery to 4.2 mm at re-entry to 4.0 mm at 5 years, decrease in mean probing pocket depth from 6.1 mm at surgery to 3.0 mm at re-entry to 3.3 mm at 5 years, and mean gingival recession from +0.4 mm at surgery to 1.0 mm at re-entry to 0.7 mm at 5 years (all at least p<0.05 from surgery to re-entry and surgery to 5 years, N.S. from reentry to 5 years via ANOVA). These favorable long-term results with CalCarb suggest that CalCarb may have a beneficial effect in the long-term clinical management of infrabony defects. PMID- 9869356 TI - Long-term stability of the mucogingival complex following guided tissue regeneration in gingival recession defects. AB - The purpose of the present study was to evaluate the stability of soft tissue conditions in gingival recession defects treated with guided tissue regeneration (GTR). The study population was selected among those patients who had been treated with GTR procedures for Miller's class I or II, deep (> or =3 mm), buccal gingival recession defects. Defects were included only when they had revealed recession depth reduction > or =2 mm and root coverage > or =60% at 6 months following GTR treatment. These defects were regarded as successfully treated and scheduled for further monitoring. 20 patients, 11 male and 9 female, aged 23 to 57 years (mean age: 33.2 years), each contributing 1 defect, were selected. 9 patients were smokers (> or =10 cigarette per day). Recession depth (RD), probing depth (PD), clinical attachment level (CAL), and width of keratinized gingiva (KG) were assessed immediately before surgery, at 6 months post-surgery (baseline examination), and at 4 years post-surgery (4-year examination). At baseline examination, RD reduction was 3.6+/-0.9 mm (mean root coverage: 80%). CAL gain amounted to 4.2+/-1.3 mm, 60% of the defects showing CAL gain > or =4 mm. KG increased from 1.9+/-1.2 mm at presurgery examination to 3.1+/-0.9 mm at baseline examination. At 4-year examination, no significant changes from baseline RD, CAL and KG recordings were observed. Differences in baseline-4 year changes between smokers and non-smokers were not statistically significant. The results of the present study demonstrate that clinical outcome achieved following GTR procedure in gingival recession defects can be maintained over periods up to 4 years. PMID- 9869357 TI - Gingival lesions in a patient with dermochondrocorneal dystrophy (Francois syndrome). A case report. AB - This report is concerned with gingival manifestations associated with a case of dermochondrocorneal dystrophy (DCCD) or Francois syndrome occurring in a 42-year old woman. Our Department treated this patient for 15 years. Oral examination of this case revealed a diffuse enlargement and severe inflammation of the attached gingiva. Systemic findings were similar to those reported in the literature for patients with DCCD. Firm papules 3 mm wide, localized on the face and on the dorsal surface of the hands, were associated with corneal involvement and progressive and severe articular disorder. Because they recurred after surgical ablation, the gingival lesions became an important problem in the management of the patient. After 10 years of unsuccessful treatment limited to scaling, oral hygiene control and mouth rinses with 0.2% chlorexidine solution, the patient was submitted to extraction of the remaining teeth, remodelling osteoplasty and cutaneous graft. An acrylic full denture was inserted. In a follow-up of 7 years, good results for the oral health of the patient were seen. PMID- 9869358 TI - Gas chromatographic-mass spectrometric determination of urinary oxoacids using O (2,3,4,5,6-pentafluorobenzyl)oxime-trimethylsilyl ester derivatization and cation exchange chromatography. AB - We introduced a new combined method to isolate, purify and quantify oxoacids in human urine. Preparation of O-(2,3,4,5,6-pentafluorobenzyl) oximes of oxoacids at pH 2 to 3 was followed by cation-exchange column chromatography for removing the biological interferences. The effluent with water was extracted with ethyl acetate and the oxoacids were quantitatively converted into their trimethylsilyl derivatives for detection by gas chromatography-mass spectrometry. Good quality control data were obtained through precision and accuracy tests. Analytical recoveries (53.5-99.8%) were quantitative for a wide variety of oxoacids. This method was used for the measurement of 18 oxoacids in the urine of healthy volunteers. PMID- 9869359 TI - Rapid quantitation of free fatty acids in human plasma by high-performance liquid chromatography. AB - We report a rapid and sensitive method for separation and quantitation of free fatty acids (FFAs) in human plasma using high-performance liquid chromatography (HPLC). Two established techniques of lipid extraction were investigated and modified to achieve maximal FFA recovery in a reasonably short time period. A modified Dole extraction method exhibited greater recovery (approximately 90%) and short processing times (30 min) compared to the method of Miles et al. Reversed-phase HPLC using UV detection was used for plasma FFA separation and quantitation. Two phenacyl ester derivatives, phenacyl bromide and p bromophenacyl bromide, were investigated in order to achieve optimal separation of individual plasma FFAs (saturated and unsaturated) with desirable detection limits. Different chromatographic parameters including column temperature, column type and elution profiles (isocratic and gradient) were tested to achieve optimal separation and recovery of fatty acids. Phenacyl bromide esters of plasma fatty acids were best resolved using an octadecylsilyl column with endcapped silanol groups. An isocratic elution method using acetonitrile-water (83:17) at 2 ml/min with UV detection at 242 nm and a column temperature of 45 degrees C was found to optimally resolve the six major free fatty acids present in human plasma (myristic [14:0], palmitic [16:0], palmitoleic [16:1], stearic [18:0], oleic [18:1] and linoleic [18:2]), with a run time of less than 35 min and detection limits in the nmol range. The entire process including plasma extraction, pre column derivatization, and HPLC quantitation can be completed in approximately 90 min with plasma samples as small as 50 microl. Over a wide physiological range, plasma FFA concentrations determined using our HPLC method agree closely with measurements using established TLC-GC methods (r2 < or = 0.95). In addition, by measuring [14C] or [3H] radioactivity in eluent fractions following HPLC separation of plasma FFA, this method can also quantitate rates of FFA turnover in vivo in human metabolic studies employing isotopic tracers of one or more fatty acids. PMID- 9869360 TI - Determination of androstenone in pig fat using packed column supercritical fluid chromatography-mass spectrometry. AB - Packed column supercritical fluid chromatography (SFC) in combination with atmospheric pressure chemical ionisation mass spectrometry was applied to the analysis of androstenone in pig fat samples. Liquefied fat samples were dissolved in dichloromethane and analysed directly by SFC without any sample purification. Chromatographic separation was achieved with a density/pressure gradient using pure carbon dioxide as the mobile phase and the analysis resulted in a quantitation limit of 0.25 microg/g with 1 microl injection volume. Good agreement was found between the SFC method and time-resolved fluoroimmunoassay by the analysis of 15 boar back fat samples. PMID- 9869361 TI - High-performance liquid chromatographic method for measuring total plasma homocysteine levels. AB - We have modified a high-performance liquid chromatographic (HPLC) procedure based on SBD-F (ammonium-7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate) pre-column derivatization to obtain an assay that is useful for routine clinical total plasma homocysteine (tHcy) analysis. The introduction of easily handled sodium borohydride instead of the traditional tri-n-butylphosphine in dimethylformamide as a reductant and a 14-min run-time using basic isocratic HPLC equipment are the more notable advantages. The addition of mercaptopropionylglycine as an internal standard contributed to improvements in the reproducibility of the assay, yielding within- and between-run precisions of 1.9 and 4% (C.V.), respectively. Reference values for fasting tHcy were 7.65+/-2.3 and 8.9+/-2.4 micromol/l, while post-methionine load gave tHcy levels of 19.9+/-5.5 and 26.8+/-5.5 micromol/l, for women and men, respectively (n=40). PMID- 9869362 TI - Determination of cellular thiols and glutathione-related enzyme activities: versatility of high-performance liquid chromatography-spectrofluorimetric detection. AB - A high-performance liquid chromatography (HPLC) method to determine the most important cellular thiols [reduced glutathione (GSH), cysteine, gamma glutamylcysteine and cysteinylglycine] is described. Separation relies upon isocratic ion-pairing reversed-phase chromatography and detection is operated by spectrofluorimetry coupled with post-column derivatization reactions using either N-(1-pyrenyl)maleimide (NPM) or ortho-phthalaldehyde (OPA). When OPA is used without co-reagent, only GSH and gamma-glutamylcysteine are detected (heterobifunctional reaction). However, either the OPA reaction in the presence of glycine in the mobile phase (thiol-selective reaction) or NPM allows the detection of all the cited thiols. The HPLC system has been validated as concerning linearity, accuracy and precision. The low detection limits reached (in the pmol range for each thiol injected) allow the screening and the quantification of thiols (as NPM derivatives) in V79cl and V79HGGT cells as well as the measurement of two cytosolic enzymes related to the glutathione synthesis, using the heterobifunctional OPA reaction. PMID- 9869363 TI - Separation of human globin chains by micellar electrokinetic capillary chromatography. AB - A new separation method of human globin chains by micellar electrokinetic capillary chromatography (MECC) is described. In this method, a 25 mM phosphate buffer (pH 2.5) containing 7 M urea and 1% (w/v) reduced Triton X-100 buffer system was used. All experiments were performed in a 47 cmx50 microm I.D. uncoated fused-silica capillary. The separation voltage was set at 19 kV. Normal globin chains derived from normal adults and newborns, alpha, beta, delta, Ggamma and Agamma globin chains as well as common variant globin chains were successfully separated within 20 min. High reproducible migration times of globin chains (CVs of intra- and inter-assay were less than 1% and 2% respectively), and quantification of Ggamma and Agamma chains (CVs for intra- and inter-assay were less than 5% and 10%, respectively) were obtained. This new MECC method provides primary information on structural modification of globin chains. It can be an important diagnostic tool in clinical laboratory practice in the field of hemoglobinopathies. PMID- 9869364 TI - Determination of adenosine and deoxyadenosine in urine by high-performance liquid chromatography with column switching. AB - The means of measurement of adenosine and deoxyadenosine in urine was developed by separating adenosine and deoxyadenosine from other compounds using high performance liquid chromatography with column switchings. This method is simple and convenient since no pretreatment of the urine is needed. Using this method, it could be demonstrated that urinary adenosine was higher in an adenosine deaminase (ADA) deficient patient who had a bone marrow transplant treatment (1.97 micromol/mmol creatinine) and in a heterozygote who had a markedly low erythrocyte ADA activity (1% of control ADA activity) (1.33 micromol/mmol creatinine) as compared to normal subjects (0.22+/-0.09 micromol/mmol creatinine, n=11). It was also noted that urinary deoxyadenosine was below the detection limits in the ADA-deficient bone marrow transplant patient, but it was detected in the heterozygote (3.7 micromol/mmol creatinine). Furthermore, it was also demonstrated that a fructose infusion increased the urinary concentration of adenosine from 0.21+/-0.03 to 2.66+/-1.21 micromol/mmol creatinine in five normal subjects. PMID- 9869365 TI - Purification of chemically synthesised dinucleoside(5',5') polyphosphates by displacement chromatography. AB - Dinucleoside(5',5') polyphosphates (ApnA, ApnG, GpnG, n=3-6) are new group of hormones controlling important biological processes. Because some of the dinucleoside(5',5') polyphosphates are commercially not available purification of chemical synthesised dinucleoside(5',5') polyphosphates became necessary in order to test their physiological and pharmacological properties. It was the aim of this study to find a method which allows purification of 0.1-0.2 g quantities of dinucleoside polyphosphates by analytical HPLC columns yielding products with impurities lower than 1.0%. Adenosine(5')-polyphospho-(5')guanosines were synthesised by mixing the corresponding mononucleotides. The reaction results in a complex mixture of ApnA, ApnG and GpnG (with n=3-6 in all cases). The reaction mixture was concentrated on a preparative C18 reversed-phase column. The concentrate was displaced on a reversed-phase stationary. As a result of displacement chromatography, anion-exchange chromatography in gradient modus yielded baseline separated dinucleoside polyphosphates (homogeneity of the fractions>99%). The identity of the substances were determined by matrix assisted laser desorption ionisation mass spectrometry. PMID- 9869366 TI - Determination of endosulfan and its metabolites in human urine using gas chromatography-tandem mass spectrometry. AB - A method was developed for determining the occupational exposure to endosulfan and its main metabolites (endosulfanether, -lactone and -sulfate) in human urine using dieldrin as an internal standard. Solid phase extraction (SPE) and gas chromatography-tandem mass spectrometry (GC-MS-MS) have been used due to their high sensitivity and selectivity in avoiding most matrix interferences. The recovery efficiencies of the tested compounds yielded more than 89.2% at the fortification level of 10 ng ml(-1) in urine and their relative standard deviations were between 9.1 and 12.8%. The detection limit of each compound ranged between 6 and 18 pg ml(-1). Urine samples from nine pest control operators were analysed and total endosulfan concentrations between 94 and 2038 pg ml(-1) were found. PMID- 9869367 TI - Quantitation of N-ethyl-3,4-methylenedioxyamphetamine and its major metabolites in human plasma by high-performance liquid chromatography and fluorescence detection. AB - A HPLC method has been developed for the analogue of Ecstasy MDE and its major metabolites N-ethyl-4-hydroxy-3-methoxyamphetamine (HME) and 3,4 methylenedioxyamphetamine (MDA) in human plasma. In the course of our investigations we found that the methylenedioxyamphetamines and HME exhibit fluorescence at 322 nm. Therefore the detection could be carried out with a fluorescence (FL) detector. Solid-phase extraction was used for sample preparation and yielded high recovery rates greater than 95%. The limit of quantitation for MDE and its metabolites in the extracts was between 1.5 and 8.9 ng/ml and the method standard deviations were less than 5%. This sensitive, rapid and reliable analytical method has been used successfully in the quantitation of the substances in plasma samples obtained from 14 volunteers in two clinical studies after p.o. administration of 100 to 140 mg MDE*HCI. The maximum plasma concentrations were 235-465 ng/ml (MDE), 67-673 ng/ml (HME) and 7-33 ng/ml (MDA), respectively. Pharmacokinetic parameters have been investigated using the plasma concentration curves. PMID- 9869368 TI - Quantification of flunitrazepam's oxidative metabolites, 3-hydroxyflunitrazepam and desmethylflunitrazepam, in hepatic microsomal incubations by high-performance liquid chromatography. AB - A high-performance liquid chromatographic assay for the quantification of the oxidative metabolites of flunitrazepam, 3-hydroxyflunitrazepam and desmethylflunitrazepam, in human liver microsomal incubations was developed. Both metabolites were quantifiable in a single assay following a solvent extraction and reversed-phase high-performance liquid chromatography with UV detection. Standard curve concentrations for both metabolites ranged from 0.2 to 10 microM. Assay performance was determined using quality control samples and the intra- and inter-day accuracy and precision as determined by the coefficient of variations which were less than 15% (0.5-6 microM) for both metabolites. This method provides good precision and accuracy for use in kinetic studies of the oxidative metabolism of flunitrazepam in human liver microsomes. PMID- 9869369 TI - Determination of pilocarpic acid in human plasma by capillary gas chromatography with mass-selective detection. AB - A novel, highly sensitive method for the determination of pilocarpic acid (PA) in human plasma is described. In addition, the method provides for the conversion of the lactone, pilocarpine (P), to PA so that a total drug presence can be determined. Using novel high-performance liquid chromatographic conditions capable of separating P, isopilocarpine (I-P), PA and isopilocarpic acid (I-PA) from each other and from endogenous plasma impurities, it was confirmed that P exclusively and quantitatively converts to PA in heparinized human plasma during storage. For the determination of PA, the selective extraction of PA from protein free plasma was accomplished using two different solid-phase extraction (SPE) cartridges in two consecutive SPE steps. After extraction, PA was lactonized with trifluoroacetic acid back to P, and both P and an internal standard were acylated using heptafluorobutyric anhydride (HFBA). The trifluoroacetylated derivatives were monitored using gas chromatography (GC) with mass spectrometric (MS) detection. This procedure allowed the sensitive and reliable determination of PA with a limit of quantification (LOQ) of 1 ng/ml, which could not be achieved using previously described methods. The assay was validated in the concentration range of 1 to 10 ng/ml with an intra-day precision (expressed as the coefficient of variation, C.V.) ranging from 9.9 to 0.5%. Inter-day precision for the quality control standard at 2.5 ng/ml showed a C.V. of 10.2%. Accuracy ranged from 94 to 102%. The assay was used to monitor the maximum systemic exposure to P, administered by the ocular route, in terms of total plasma PA (P and PA). PMID- 9869370 TI - Determination of N,N',N"-triethylenethiophosphoramide and its active metabolite N,N',N"-triethylenephosphoramide in plasma and urine using capillary gas chromatography. AB - A sensitive assay for the determination of N,N',N"-triethylenethiophosphoramide (thioTEPA) and its metabolite N,N',N"-triethylenephosphoramide (TEPA) in micro volumes human plasma and urine has been developed. ThioTEPA and TEPA were analysed using gas chromatography with selective nitrogen-phosphorus detection or mass spectrometry after extraction with a mixture of 1-propanol-chloroform from the biological matrix. Diphenylamine was used as internal standard. The limit of detection was 1.5 ng/ml for thioTEPA and 2.5 ng/ml for TEPA, using 100 microl of biological sample; recoveries ranged between 70 and 90% and both accuracy and precision were less than 10%. Linearity was accomplished in the range of 10-1000 ng/ml for plasma and 100-10000 ng/ml for urine using thermionic nitrogen phosphorus detection. With mass spectrometry a linear range of 100-25000 ng/ml TEPA in plasma or urine was obtained. For thioTEPA a second-order polynomial function describes the relationship between the analyte concentration in the range of 500-25000 ng/ml and detection response. TEPA proved to be stable in plasma and urine for at least 10 weeks at -80 degrees C. ThioTEPA and TEPA plasma concentrations of two patients treated with thioTEPA are presented demonstrating the applicability of the assay for clinical samples. PMID- 9869371 TI - Sensitive and specific liquid chromatographic-tandem mass spectrometric assay for barnidipine in human plasma. AB - A sensitive and specific LC-MS-MS assay has been developed and validated for barnidipine (1-benzyl-3-pyrrolidinyl)methyl-2,6-dimethyl-4(m-nitrophenyl)-1,4 dihydr opyridine-3,5-dicarboxylate). The assay involves a simple and rapid solid phase extraction procedure. Sample analysis was on a Spherisorb S3ODS2 100 mmX2 mm I.D. column, with a Finnigan TSQ 7000 mass spectrometer, using an electrospray interface and selective reaction monitoring (SRM). The intra- and inter-day precision and accuracy, determined as the coefficient of variation and relative error, respectively, were 11.8% or less. The limit of quantitation was 0.03 ng/ml, and the calibration was linear between 0.03 and 3.0 ng/ml. The method has been used successfully for the measurement of over two thousand human plasma samples from pharmacokinetic clinical trials. PMID- 9869372 TI - Determination of ivabradine and its N-demethylated metabolite in human plasma and urine, and in rat and dog plasma by a validated high-performance liquid chromatographic method with fluorescence detection. AB - A sensitive and selective high-performance liquid chromatographic method with native detection of fluorescence was developed and validated for the quantitation of ivabradine and its N-demethylated metabolite in plasma (rat, dog, human) and human urine. The procedure involves the use of an analogue as internal standard, solid-phase extraction on cyano cartridges, separation on a Nova-Pak C8 column and fluorescence detection. Calibration curves are linear in the concentration ranges from 0.5 to 100 ng/ml in plasma and 2.0 to 500 ng/ml in urine with a limit of quantitation set at 0.5 and 2.0 ng/ml in plasma and urine, respectively. The analysis of plasma and urine samples (spiked with the analytes at low, medium and high concentrations of the calibration range) demonstrates that both analytes can be measured with precision and accuracy within acceptable limits. Quality controls spiked with analyte concentrations up to 10000 ng/ml can also be analysed with excellent precision and accuracy after dilution of the samples. The parent drug and its metabolite are stable in plasma and urine after short-term storage (24 h at room temperature and after three freeze-thaw cycles) as well as after long-term storage at -20 degrees C (at least 6 months in animal plasma and 12 months in human plasma and urine). The method has been used to quantify both compounds in plasma and urine samples from clinical and non-clinical studies with ivabradine. PMID- 9869373 TI - High-performance liquid chromatographic separation and nanogram quantitation of bupivacaine enantiomers in blood. AB - Chiral separation of rac-bupivacaine extracted from blood was achieved with similar limits of detection but using a much simpler sample preparation than reported previously. The simple one-step sample preparation devised was highly robust and efficient and allowed a very high throughput of samples. The high performance liquid chromatography (HPLC) conditions used gave baseline separation of the enantiomers with high sensitivity. R-(+)-bupivacaine and S-(-)-bupivacaine blood concentrations were determined using a chiral stationary phase (AGP, ChromTech) with diode array detection at 220 nm; this wavelength produced a stable baseline allowing semi-automated analysis. Sample preparation involved addition of internal standard (diphenhydramine), basification of blood, extraction with n-hexane, concentration of the extract to dryness and reconstitution in 0.002 M phosphoric acid. At rac-bupivacaine concentrations of 0.5, 5 and 50 microg/ml in blood, assay accuracy as estimated by coefficients of variation (C.V.s), were 3.3, 1.4, and 1.6%, respectively, for R-(+)-bupivacaine and 3.7, 2.0 and 1.5%, respectively, for S-(-)-bupivacaine. Using 0.6-ml samples, the estimated limits of detection for R-(+)-bupivacaine and S-(-)-bupivacaine were both 15 ng/ml of blood. Calibration curves (n=188) were linear from 0.1 to 50 microg/ml with all correlation coefficients being greater than 0.99. This semi automated method was applied to studies involving whole body pharmacokinetics with intravenous doses ranging from 12.5 to 350 mg and regional myocardial pharmacokinetics with coronary arterial doses ranging from 2.5 to 12.5 mg. These studies generated approximately 12000 blood samples. PMID- 9869374 TI - Improved method for the simultaneous determination of proguanil and its metabolites by high-performance liquid chromatography and solid-phase extraction of 100-microl capillary blood samples dried on sampling paper. AB - An improved method is presented for the determination of proguanil, cycloguanil and 4-chlorophenylbiguanide in 100-microl capillary blood samples applied to sampling paper. This method also utilises a solid-phase extraction technique and high-performance liquid chromatography. Different kinds of sampling paper, such as ion-exchange and cellulose sampling paper were tested. The best elution recovery (70-80%) was obtained after treatment of cellulose sampling paper with a quaternary ammonium compound. The limit of determination was 50 nmol/l for cycloguanil and 4-chlorophenylbiguanide and 125 nmol/l for proguanil using 100 microl capillary blood. The stability of the analytes and elution performance from sampling paper was validated at different temperature and storage time. Venous blood and capillary blood concentrations of proguanil and metabolites were found to be similar. PMID- 9869375 TI - Determination of ceftazidime in plasma using high-performance liquid chromatography and electrochemical detection. Application for individualizing dosage regimens in elderly patients. AB - This study describes a sensitive HPLC-electrochemical detection method for the analysis of ceftazidime, a third-generation cephalosporin, in human plasma. The extraction procedure involved protein precipitation with 30% trichloroacetic acid. The separation was achieved on a reversed-phase column (250X4.6 mm I.D., 5 microm) packed with C18 Kromasil with isocratic elution and a mobile phase consisting of acetonitrile-25 mM KH2PO4-Na2HPO4 buffer, pH 7.4 (10:90, v/v). The proposed analytical method is selective, reproducible and reliable. The assay has a precision of 0.2-15.1% (C.V.) in the range of 5-200 microg mil(-1). (corresponding to 0.5 to 20 ng of ceftazidime injected onto the column), and is optimised for assaying 50 microl of plasma. The extraction recovery from plasma was approximately 100%. The method was highly specific for ceftazidime and there was no interference from either commonly administered drugs or endogenous compounds. This assay was used to measure ceftazidime in elderly patients for therapeutic drug monitoring. PMID- 9869376 TI - Simultaneous quantitative determination of the HIV protease inhibitors amprenavir, indinavir, nelfinavir, ritonavir and saquinavir in human plasma by ion-pair high-performance liquid chromatography with ultraviolet detection. AB - A reversed-phase high-performance liquid chromatographic assay for the simultaneous quantitative determination of five HIV protease inhibitors (i.e. amprenavir, indinavir, nelfinavir, ritonavir, and saquinavir) in human plasma is described. Sample pretreatment consisted of solid-phase extraction prior to ion pair, reversed-phase high-performance liquid chromatography with ultraviolet detection at 210 nm (amprenavir, indinavir and nelfinavir) and 239 nm (saquinavir and ritonavir). For amprenavir, indinavir and saquinavir the method has been validated over the range of 25 ng/ml to 25 microg/ml using a 0.6 ml sample volume. For nelfinavir and ritonavir the method has been validated over the range of 50 ng/ml to 25 microg/ml. The method proved to be accurate, with an average accuracy at four concentrations ranging from 90.6 to 109.2%, and precise, with the within-day and between-day precision ranging from 1.8 to 6.7%, and 0.7 to 7.6%, respectively. The protease inhibitors which can be quantified by using this assay proved to be stable under various conditions. This assay can readily be used in a hospital laboratory for the routine monitoring of plasma concentrations of these protease inhibitors. PMID- 9869377 TI - Determination of the antitumor agent depsipeptide in plasma by liquid chromatography on serial octadecyl stationary phases. AB - A high-performance liquid chromatographic assay has been developed and validated for the determination of the antitumor agent depsipeptide (FR-901228) in plasma samples from patients with advanced cancer. After the plasma proteins were precipitated with acetonitrile, the supernatant was extracted with ethylacetate. Depsipeptide was chromatographed on two serial octadecylsilica stationary phases using a mobile phase consisting of acetonitrile-potassium phosphate buffer (0.03 M, pH 3) (27:73, v/v), at a flow-rate of 2.0 ml/min and at ambient temperature. The method was linear over a 50 to 2000 ng/ml range and the intra- and inter-day coefficients of variations were less than 8%. The method was applied to the determination of the plasma concentration-time profile for 14 patients with advanced cancer receiving from 1 to 7.5 mg/m2 of depsipeptide per day as a continuous 4-h infusion. PMID- 9869378 TI - High-performance liquid chromatographic determination of erdosteine and its optical active metabolite utilizing a fluorescent chiral tagging reagent, R-(-)-4 (N,N-dimethylamiosulfonyl)-7-(3-aminopyrrolidin-1-yl)-2 ,1,3-benzoxadiazole. AB - Chiral separation of racemic M1 metabolized from erdosteine was investigated by reversed-phase chromatography. The sensitive determination of M1 and erdosteine with UV detection was difficult because of their low absorptivity in the effective wavelength region. To improve the sensitivity and separatability, one thiol and two carboxyl groups in the M1 structure were labelled with DBD-F and R (-)-DBD-APy, respectively. Non-fluorescent DBD-F quantitatively reacted with thiol in M1 at room temperature for 30 min in borate buffer (pH 9.3) to produce the fluorescent derivative. On the other hand, the labelling of two carboxyls was carried out with a chiral fluorescent reagent, R-(-)-DBD-APy, in acetonitrile containing DPPA. The derivatives corresponding to a pair of the enantiomers were completely separated with water-acetonitrile containing 0.1% TFA as the mobile phase by an ODS column. Erdosteine with a carboxyl group was also labelled with R (-)-DBD-APy and separated together with M1 derivatives. The detection limits (S/N=3) of erdosteine and M1 were 0.37 and 0.22 pmol, respectively. The proposed derivatization and separation methods were applied to simultaneous determination of racemic M1 and erdosteine in rat plasma after administration of erdosteine. The amounts of both enantiomers of M1 were essentially the same in oral and intravenous administrations. In contrast, total amounts (reduced-form and oxidized-form) of S-(-)-M1 in rat plasma were higher than those of R(+)-M1 in both administrations. PMID- 9869379 TI - Determination of PNU 157706, a new dual inhibitor of 5alpha-reductase, in rat plasma by high-performance liquid chromatography with ultraviolet detection. AB - A HPLC procedure was developed and validated for determining nanogram per milliliter concentrations of the dual 5alpha-reductase inhibitor PNU 157706 in rat plasma. The compound was extracted from plasma with diethyl ether followed by purification using a CN cartridge. The chromatographic separation was performed with a C18 column using a water-acetonitrile-methanol mixture as eluent. UV detection at 210 nm was used for the quantification of the compound over the concentration range 5-500 ng/ml plasma. The method has a lower limit of quantification of 5 ng/ml and good precision and accuracy. This method has performed well during analysis of several toxicokinetic and pharmacokinetic studies in the rat. PMID- 9869380 TI - At-line solid-phase extraction for capillary electrophoresis: application to negatively charged solutes. AB - The analysis of complex biological samples with capillary electrophoresis (CE) requires proper sample pretreatment. In this paper the applicability of solid phase extraction (SPE) coupled at-line with CE is studied, by using a laboratory made interface. A fresh (disposable) SPE cartridge is used for each sample to prevent carry-over effects. The sample handling procedure is performed parallel with the analysis of the previous sample, to improve sample throughput. Using this set-up, negatively charged test compounds (some non-steroid anti inflammatory drugs) can be determined in serum and urine. The method is linear over at least two decades and detection limits are around 40 microg/l. A single capillary, flushed only once a week with a sodium hydroxide solution, was used without problems for the analysis of ca. 900 samples during 1 year. The robustness of the system was very good: no blocking of loop, interface or capillary was found during this period. Furthermore, the system was successfully used for overnight runs. PMID- 9869381 TI - Determination of acrolein in human urine by headspace gas chromatography and mass spectrometry. AB - A rapid and sensitive headspace gas chromatographic and mass spectrometric (GC MS) method was developed for the determination of acrolein in human urine. A 0.5 ml urine sample in a glass vial containing propionaldehyde as an internal standard was heated at 80 degrees C for 5 min. A 0.1-ml volume of headspace vapor was injected into a GC-MS instrument. Acrolein and propionaldehyde were coeluted at 3.1 min using a DB-1 capillary column, and well separated by selective ion monitoring (SIM) mode using ions m/z 56.05 and m/z 58.05. The interassay and intraassay coefficient of variation were 0.99% and 3.3%. The calibration curve demonstrated a good linearity throughout concentrations ranging from 1 to 1000 nM. However, due to a wide variation of acrolein evaporation rates from human urine, a calibration curve must be established for each urine specimen using a standard addition method and detection limit varied from 1 to 5 nM. The total analysis time for two samples from one urine specimen required about 15 min. Therefore, this method is convenient for the urgent monitoring of urinary acrolein in patients to whom alkylating agents are administered. PMID- 9869382 TI - Quantitative analysis of acetaldehyde in whole blood from human and various animals by gas chromatography. AB - Acetaldehyde present in the blood of bull, chicken, hamster, horse, human, monkey, pig, rabbit, rat and sheep, was quantitatively analyzed by a newly developed gas chromatographic method. Acetaldehyde in a blood sample was reacted with cysteamine to give 2-methylthiazolidine, which was extracted with dichloromethane and subsequently analyzed by gas chromatography with a fused silica capillary column and a nitrogen-phosphorus detector. The quantities of acetaldehyde found in blood ranged from 2.04 micromol/ml (hamster) to 14.8 micromol/ml (pig). The quantity of acetaldehyde recovered from human blood was 6.17 micromol/ml. PMID- 9869383 TI - Determination of free acetaldehyde in total blood for investigating the effect of aspartate on metabolism of alcohol in mice. AB - To explore the effect of sodium L-aspartate monohydrate (aspartate) as a NAD+ regenerating agent for acetaldehyde in alcohol metabolism, a simple HPLC method has been developed for the measurement of free acetaldehyde in total mice blood digested with alcohol and aspartate. The blood samples were collected in EDTA Vacutainer tubes, and treated with 2,4-dinitrophenylhydrazine (DNP hydrazine) reagent in total blood. Acetaldehyde DNP hydrazone was extracted from total blood and analyzed by HPLC using an Ultrasphere ODS column. The compounds were separated using acetonitrile-water (50:50, v/v) as mobile phase and detected at 356 nm. The detection limit for acetaldehyde DNP hydrazone was 0.1 ppm. A blank determination was carried out for each analysis and subtracted from the results. The amount of acetaldehyde in blood has been determined as a function of time lapse after sole alcohol administration and aspartate ingestion followed by alcohol administration, respectively. This comparative analysis demonstrates that the ingestion of aspartate before the administration of alcohol dramatically decreases the aldehyde level in blood, and aspartate may be utilized as a prospective antagonist for acceleration of ethanol metabolism and prevention of acetaldehyde toxicity. PMID- 9869384 TI - Validated high-performance liquid chromatographic assay for the determination of promazine in human plasma. Application to pharmacokinetic studies. AB - A high-performance liquid chromatographic method for the determination of promazine in human plasma is described. The assay involves a single-step liquid liquid extraction using pentane-2-propanol (98:2, v/v). The analyte of interest and the internal standard chlorpromazine were separated on a Spherisorb CN column using a mobile phase of acetonitrile-50 mM ammonium acetate (9:1, v/v). Electrochemical detection was achieved using an applied potential of +750 mV. The assay was validated according to international requirements prior to application to a pharmacokinetic study and was found to be specific, accurate and precise with a linear range of 0.25-25 ng ml(-1). PMID- 9869385 TI - Improved selectivity for high-performance liquid chromatographic determination of clonazepam in plasma of epileptic patients. AB - We report a high-performance liquid chromatography method for clonazepam determination in plasma. The use of a synthetic silica-based stationary phase markedly improved clonazepam resolution compared to standard reversed-phase columns. A liquid-liquid extraction was used, associated with reversed-phase chromatography, gradient elution and ultraviolet detection. Accuracy and precision were satisfactory at therapeutic concentrations. Selectivity was studied for benzodiazepines or other antiepileptic drugs, with particular attention to newly marketed drugs i.e., gabapentine and vigabatrin. No interfering substance was evidenced. Under the conditions described, it was possible to quantify clonazepam at nanogram level even when carbamazepine was present at therapeutic concentrations. PMID- 9869386 TI - Analysis of the enantiomers of citalopram and its demethylated metabolites using chiral liquid chromatography. AB - A procedure using a chirobiotic V column is presented which allows separation of the enantiomers of citalopram and its two N-demethylated metabolites, and of the internal standard, alprenolol, in human plasma. Citalopram, demethylcitalopram and didemethylcitalopram, as well as the internal standard, were recovered from plasma by liquid-liquid extraction. The limits of quantification were found to be 5 ng/ml for each enantiomer of citalopram and demethylcitalopram, and 7.5 ng/ml for each enantiomer of didemethylcitalopram. Inter- and intra-day coefficients of variation varied from 2.4% to 8.6% for S- and R-citalopram, from 2.9% to 7.4% for S- and R-demethylcitalopram, and from 5.6% to 12.4% for S- and R- didemethylcitalopram. No interference was observed from endogenous compounds following the extraction of plasma samples from 10 different patients treated with citalopram. This method allows accurate quantification for each enantiomer and is, therefore, well suited for pharmacokinetic and drug interaction investigations. The presented method replaces a previously described highly sensitive and selective high-performance liquid chromatography procedure using an acetylated 3-cyclobond column which, because of manufactural problems, is no longer usable for the separation of the enantiomers of citalopram and its demethylated metabolites. PMID- 9869387 TI - High-performance liquid chromatographic determination of ropivacaine, 3-hydroxy ropivacaine, 4-hydroxy-ropivacaine and 2',6'-pipecoloxylidide in plasma. AB - A sensitive HPLC method has been developed for the determination of ropivacaine, 3-hydroxy-ropivacaine, 4-hydroxy-ropivacaine and 2',6'-pipecoloxylidide in plasma. The procedure involved extraction from plasma with a mixture of n-heptane ethyl acetate and a back-extraction into an acidified aqueous solution. The chromatography was achieved using a LiChrospher RPB C8 column with a mobile phase consisting of a mixture of acetonitrile and pH 2.1, 0.01 M potassium dihydrogenphosphate, the latter phase containing 0.005 M 1-heptanesulfonic acid for ropivacaine metabolites analysis. The extraction yields of ropivacaine, 3 hydroxy-ropivacaine, 4-hydroxy-ropivacaine and 2',6'-pipecoloxylidide were 94.7%, 79.4%, 79.4% and 77.7%, respectively. The limits of detection of ropivacaine, 3 hydroxy-ropivacaine, 4-hydroxy-ropivacaine and 2',6'-pipecoloxylidide in plasma samples were 0.9 ng/ml, 3 ng/ml, 5 ng/ml and 1 ng/ml, respectively. PMID- 9869388 TI - Alternative method for determination of ceftazidime in plasma by high-performance liquid chromatography. AB - A high-performance liquid chromatography procedure was developed to analyze ceftazidime concentrations in plasma. The procedure consisted of solid phase extraction followed by ion-pairing reverse-phase chromatography. An excellent linear relationship between ceftazidime peak height measurements and concentrations was demonstrated over the concentration range of 1-200 microg ml( 1). The advantage of this assay is the elimination of interference at the ceftazidime elution time that has been noted in previous studies and in our experience. Thus, this study describes an alternative, simple methodology that is clinically useful for analyzing ceftazidime in the research setting. PMID- 9869389 TI - High-performance liquid chromatographic bio-analysis of PSC 833 in human and murine plasma. AB - We have developed a rapid, sensitive and selective method for the determination of the cyclosporin analog PSC 833 in human and mouse plasma using cyclosporin A as internal standard. The assay uses liquid-liquid extraction with diethyl ether for sample clean-up followed by reversed-phase high-performance liquid chromatography with UV detection at 210 nm. Good peak shapes were obtained using a NovaPak Phenyl column operating at 72 degrees C. Good selectivity from endogenous compounds was achieved using a mobile phase composed of methanol acetonitrile-water (34:34:32). The retention times of cyclosporin A and PSC 833 were approximately 7.8 and 11.7 min, respectively, with two major endogenous peaks at 9.2 and 16.7 min. Selective decreasing of the retention times of cyclosporin A and PSC 833 relative to these interferences occurring upon aging of the column was balanced by increasing the percentage of methanol relative to acetonitrile. No other late eluting peaks were present, resulting in a total analysis time of 20 min per sample. The assay performance in human plasma was good. The absolute recovery of PSC 833 after the sample clean-up step was 48+/ 6%. The lower limit of quantitation was 0.05 microM using 500 microl of sample. Within the linear dynamic range of the assay (0.10-5.0 microM) the accuracy was close to 100% and within-day and between-day variation less than 7%. Because of the limited availability of blank mouse plasma, the concentration in samples from mice were determined using calibration curves constructed in human plasma. The lower limit of quantitation in mouse was 0.25 microM using 200 microl of sample. Overall, the performance of the assay in mouse plasma was somewhat less than in human plasma but accuracy and precision were within the ranges that are considered acceptable for bio-analytical assays. PMID- 9869390 TI - Plasma-kallikrein clearance during liver regeneration after partial hepatectomy in the rat. AB - AIMS/BACKGROUND: The liver clears circulating plasma-kallikrein through a receptor-mediated endocytosis process: an initial fast phase is followed by a slow exponential phase. METHODS: To determine whether the clearance rate of plasma-kallikrein is affected during liver regeneration, we perfused isolated rat livers with rat plasma-kallikrein (rPK) at 0, 1, 2, 3 and 7 days after partial hepatectomy or sham operation. RESULTS: Liver regeneration was followed by the expression of the proliferating-cell nuclear antigen (PCNA) labeling index. The serum concentration of alpha2-macroglobulin, an acute phase protein in rats, was measured. At day 1, the fast phase of rPK clearance rate increased in hepatectomized rats when compared with day 0 (4.9+/-0.4 and 3.7+/-0.4 mU/g liver min, p<0.05). However, at day 2, the rPK fast phase clearance rate dropped significantly (2.6+/-0.2, p<0.05), when compared with day 1. No difference was found among the sham groups at different days of hepatectomy. These changes seem to be independent of the acute phase reaction. The regenerative liver weight increased continuously during the observation period. PCNA expression increased significantly after hepatectomy, with maximal PCNA-labeling indices at days 1 and 2, declining thereafter. CONCLUSION: The rPK fast phase clearance rate changes during liver regeneration, with a zenith occurring when PCNA labeling index is maximal (day 1) and a nadir occurring at the mitotic phase (day 2). PMID- 9869391 TI - Comparative study of 5' UTR and NS3R primers for the detection of GB virus C/hepatitis G virus RNA in Japanese. AB - AIMS/BACKGROUND: Many epidemiological studies of new hepatitis viruses, including GB virus C (GBV-C) and hepatitis G virus (HGV), have used polymerase chain reaction (PCR) primers designed for the third nonstructural region (NS3R). However, a homology study of GBV-C and HGV genomes revealed that the 5' untranslated region (5'UTR) was more conserved than NS3R. METHODS: We attempted to detect GBV-C/HGV using PCR primers corresponding to the 5' UTR, and compared its incidence to that derived from NS3R primers. Furthermore, PCR products amplified using the 5' UTR primers were sequenced and subjected to phylogenetic analysis. RESULTS: In patients with chronic hepatitis C, the prevalence of GBV C/HGV by PCR with the NS3R and 5' UTR primers was 5.1% (4/78) and 17.9% (14/78), respectively, and in patients on hemodialysis, it was 0% (0/81) and 5.9% (5/85), respectively. We could not detect GBV-C/HGV in patients with non-A-C liver disease. The incidence of GBV-C/HGV by 5' UTR primers was higher than by NS3R primers. After DNA sequencing at 5' UTR, phylogenetic analysis showed two types of GBV-C/HGV, Jap and HGV types. CONCLUSION: 5' UTR primers proved highly sensitive for detection of GBV-C/HGV and were superior to the NS3R primers. PMID- 9869392 TI - Areas of sinusoidal surface hepatocyte nuclear predominance in type C chronic hepatitis. AB - AIMS/BACKGROUND: Thick hepatic plates have been considered one of the morphological characteristics of hepatocyte regeneration in cirrhotic nodules. They can be recognized by the sinusoidal surface predominance of their nuclei. We have investigated the prevalence of this in HBV and HCV infections. METHODS AND RESULTS: This feature was more frequently present in type C chronic hepatitis with low activity of inflammation and low grade of fibrosis, than with type B chronic hepatitis. Additionally, this area of sinusoidal surface hepatocyte nuclear predominance (ASSHNP) was seen in zone II, rather than in periportal zones, in type C chronic hepatitis. Clinical data were analyzed statistically. Immunohistochemical reactivity of type IV collagen, laminin, Ulex europaeus agglutinin 1 lectin (UEA-1), and factor VIII-related antigen were increased in ASSHNP. Immunohistochemical staining of Ki-67 antigen was performed in order to assess the regenerative capacity of this area and showed a low level of regeneration. Ultrastructure of this area in type C chronic hepatitis showed a decrease in the number of mitochondria and an increase of nuclear pleomorphism together with basement membrane formation in the space of Disse. CONCLUSION: Although the cause of these abnormalities was not clarified in this study, it is suggested that they are related to chronic hepatitis C virus (HCV) infection per se, rather than regeneration or inflammatory activity. These changes may be significant in HCV-associated hepatocarcinogenesis. PMID- 9869393 TI - Perforin and granzyme B lytic protein expression during chronic viral and autoimmune hepatitis. AB - BACKGROUND: Cytotoxic T lymphocytes and Kupffer cells are essential components of the immune response during liver diseases. Recent studies have highlighted the role of cytotoxic T lymphocytes using Fas and its ligand in induced hepatocyte death during acute and chronic hepatitis. METHODS: In the present work, the main purpose was to investigate perforin and granzyme B expression in liver biopsies of patients with chronic hepatitis (10 HBV, 14 HCV and 10 autoimmune hepatitis) using immunohistochemistry. The liver biopsies of two normal individuals were also studied in the same conditions. RESULTS: Few intrahepatic T lymphocytes expressed perforin and granzyme B, while a large number of Kupffer cells were positive for both proteins in all the patients tested. The co-localization of perforin and granzyme B, and CD3 or CD68 antigens was visualized, respectively, in T cells and Kupffer cells, using confocal microscopy. In situ hybridization assays confirmed that perforin and granzyme B mRNAs were present in the liver during chronic hepatitis. The results were similar among the three groups of patients and whatever the activity of the disease. Perforin and granzyme B expression was lacking in liver samples from normal individuals. CONCLUSIONS: These data suggest a minor role for the T cell-mediated perforin/granzyme B death pathway, and a putative role for Kuppfer cells via lytic protein release, during chronic hepatitis. PMID- 9869394 TI - Mature hepatocytes actively divide and express gamma-glutamyl transpeptidase after D-galactosamine liver injury. AB - AIMS/BACKGROUND: We studied the fate of hepatocytes in the rat liver after D galactosamine injury by genetic labeling using recombinant retroviruses carrying the Escherichia coli lacZ gene coupled to a nuclear localization signal. METHODS: Hepatocytes were either labeled by direct injection of 2.5 ml high-titer retrovirus-containing medium in the regenerating liver parenchyma after administration of a single dose of D-galactosamine. Alternatively hepatocytes were pre-labeled, 24 h after a two-thirds hepatectomy, by injecting the same volume of retroviral solution in the portal vein and D-galactosamine was administered 15 days later. Gamma-glutamyl transpeptidase and beta-galactosidase activities were assessed on cryostat sections, along with localization of the hepatocyte-specific HES6 antigen. RESULTS: Morphological observations, as well as beta-galactosidase activity detection, showed that hepatocytes actively divide as early as 1 day after D-galactosamine injection. Gamma-glutamyl transpeptidase activity was detected in biliary cells, but also in mature hepatocytes, pre labeled with beta-galactosidase before D-galactosamine administration. CONCLUSIONS: These experiments demonstrate that hepatocytes can divide to restore the liver mass after D-galactosamine liver injury. Furthermore, we also show that gamma-glutamyl transpeptidase, which has been reported to be expressed only by fetal or preneoplastic hepatocytes, can be re-expressed by mature hepatocytes during the recovery process. PMID- 9869395 TI - Quantitative and functional analysis of core-specific T-helper cell and CTL activities in acute and chronic hepatitis B. AB - AIMS/BACKGROUND: CD4+ T-helper cell (Th) responses to hepatitis B virus (HBV) core antigen (HBc) are increased during exacerbations in acute and chronic hepatitis B (AHB, CHB) and might influence the induction of CD8+ cytotoxic T lymphocytes (CTL) that are important for viral clearance. METHODS: HBc-specific proliferative responses and cytokine release of blood mononuclear cells (PBMC) were studied in patients with AHB or CHB, as well as responders and non responders to interferon-alpha treatment (IFN-R, IFN-NR), by [3H]-thymidine uptake, enzyme-linked immunosorbent assay (ELISA) and Elispot assay and were compared to peptide HBc18 27-specific CTL precursor frequencies among CD8+ T cells derived from HLA-A2+ patients. RESULTS: HBc-specific proliferative PBMC responses and Th frequencies were significantly increased in AHB patients compared with untreated CHB patients. PBMC derived from IFN-R showed stronger cellular responses than IFN-NR. Stimulated PBMC from all patient groups secreted significantly more IFN-gamma than IL-4 indicating Th1/Th0 cell responses. Furthermore, in AHB and IFN-R patients, high peptide HBc18-27-specific CTL precursor frequencies closely correlated with strong HBc-specific Th responses, whereas in untreated CHB and IFN-NR patients lower CTL frequencies were observed without correlation to Th activities. CONCLUSIONS: HBV core-specific Th-cell responses appeared to support efficient CTL induction in patients with viral clearance, whereas in chronic HBV carriers quantitatively insufficient Th and CTL responses were observed. This observation could be important for future therapeutic strategies. PMID- 9869396 TI - Stromal expression of urokinase-type plasminogen activator receptor (uPAR) is associated with invasive growth in primary liver cancer. AB - AIMS/BACKGROUND: Expression of urokinase-type plasminogen activator receptor (uPAR) was studied in 25 hepatocellular carcinomas (HCCs) and seven cholangiocellular carcinomas (CCCs) by immunohistochemistry. METHODS AND RESULTS: uPAR was expressed mostly by host cells distributed along the tumour-host interface in all cases of HCC and CCC, and its expression was higher in CCC. These uPAR-positive cells were identified as macrophages by observation of serial sections stained for CD68, a marker for macrophages. Cancer cells were positive for uPAR in only one case of poorly differentiated HCC with sarcomatous changes and in three cases of CCC. Hepatocellular carcinomas were classified into two types: those with a fibrous capsule (expansive type) and those without a fibrous capsule (invasive type). Invasive-type HCCs showed more prominent expression of uPAR by macrophages than expansive HCCs (p<0.001), to approximately the same degree as that of CCC. Extrahepatic metastasis was observed in two of 16 expansive HCCs, five of nine invasive HCCs and six of seven CCCs. CONCLUSIONS: Our findings suggest that uPAR expression mainly by macrophages is associated with invasive growth of cancer cells into the surrounding tissue in primary carcinoma of the liver. PMID- 9869397 TI - Noninvasive assessment of spontaneous baroreflex sensitivity in patients with liver cirrhosis. AB - AIMS/BACKGROUND: An impairment of baroreceptor sensitivity has been found in liver cirrhosis. Noninvasive and spontaneous estimates of baroreflex sensitivity are obtained from beat-to-beat blood pressure and heart rate recordings by means of cross-spectrum analysis and calculation of alpha-index (as a measure of baroreflex gain). The aim of the present study was to investigate the function of the spontaneous baroreflex sensitivity related to clinical Child score in liver cirrhosis. METHODS: The alpha-index was evaluated in 40 cirrhotic patients (18 with and 22 without ascites) and 17 healthy subjects by analysing finger arterial pressure recorded noninvasively with the Portapres device. RESULTS: Baroreflex sensitivity was significantly lower in cirrhotic patients with and without ascites compared with healthy subjects (p<0.01). Furthermore, in patients with ascites the baroreflex gain was significantly related to plasma sodium (p<0.01). A significant inverse relationship was present between baroreflex gain and grade of Child score and the severity of ascites (p<0.01). There were no significant relationships between hormonal parameters (catecholamines, renin, aldosterone, arginine-vasopressin, atrial natriuretic peptide and nitric oxide) and baroreflex gain. No significant differences were found between healthy subjects and cirrhotic patients with respect to systolic and diastolic blood pressure total variability in a supine position, whilst it was lower in cirrhotic patients with ascites in a tilted position (p<0.05). CONCLUSION: Our findings showed that baroreflex sensitivity was significantly impaired in cirrhotic patients when compared with healthy subjects. In addition, there was a significant trend toward lower baroreflex sensitivity values with the grade score of Child class (p<0.01). Spectral analysis of the alpha-index provides viable alternatives to the pharmacological approach for estimation of baroreflex sensitivity and may represent a prognostic tool to identify cirrhotic patients at increased risk of adverse events. PMID- 9869398 TI - Power Doppler sonography for hepatocellular carcinoma: factors affecting the power Doppler signals of the tumors. AB - AIMS/BACKGROUND: A recent advancement in Doppler ultrasonography (US) is power Doppler for detecting low-velocity blood flow at the microvascular level with angle independence. The present study was performed to characterize the factors contributing to the power Doppler signals of hepatocellular carcinoma (HCC). METHOD: Correlation of Doppler signals of HCC in 114 patients with 178 HCC nodules was analyzed in relation to the findings of CT and angiography, tumor characteristics (size, echo pattern, and histological differentiation of tumor), viral markers and severity of liver disease. RESULTS: The sensitivity of power Doppler US was superior to that of CT and angiography (each p<0.05; McNemar's test). The detection rate of power Doppler signal was significantly higher in tumors with diameter > or =2 cm (vs <2 cm in diameter), and with low/mixed echo pattern (vs high echo appearance), and with moderately/poorly differentiated HCC (vs well-differentiated HCC). Univariate analysis revealed that echo pattern, tumor size and histological differentiation of HCC in addition to CT and angiographic findings were significant. Multivariate analysis showed that tumor size and differentiation were significant. CONCLUSION: These results indicate that tumor characteristics play an important role in power Doppler signals and that these could be assessed by the presence or absence of power Doppler signals. PMID- 9869399 TI - Protein phosphatase 2C (PP2C) function in higher plants. AB - In the past few years, molecular cloning studies have revealed the primary structure of plant protein serine/threonine phosphatases. Two structurally distinct families, the PP1/PP2A family and the PP2C family, are present in plants as well as in animals. This review will focus on the plant PP2C family of protein phosphatases. Biochemical and molecular genetic studies in Arabidopsis have identified PP2C enzymes as key players in plant signal transduction processes. For instance, the ABI1/ABI2 PP2Cs are central components in abscisic acid (ABA) signal transduction. Arabidopsis mutants containing a single amino acid exchange in ABI1 or ABI2 show a reduced response to ABA. Another member of the PP2C family, kinase-associated protein phosphatase (KAPP), appears to be an important element in some receptor-like kinase (RLK) signalling pathways. Finally, an alfalfa PP2C acts as a negative regulator of a plant mitogen-activated protein kinase (MAPK) pathway. Thus, the plant PP2Cs function as regulators of various signal transduction pathways. PMID- 9869400 TI - Arabidopsis thaliana contains a large family of germin-like proteins: characterization of cDNA and genomic sequences encoding 12 unique family members. AB - We have identified 39 Arabidopsis thaliana ESTs encoding germin-like proteins (GLPs) and have completely sequenced 25 of these cDNAs. Our analysis demonstrates that the Arabidopsis genome contains a gene family with at least 12 GLP genes. Comparisons with other known germins and germin-like proteins indicate that these Arabidopsis GLP subfamilies are unique from wheat germin. All other known GLPs fall into one of these subfamilies. The translated GLPs show approximately 35% amino acid identity with other GLPs outside of their subfamily and significantly higher levels of identity within their respective subfamily. The 3' ends of many of the GLP cDNAs are heterogeneous and several sites of polyadenylation are used. Ten of the GLPs have N-terminal signal sequences and most appear to be exported from the cell. Structurally, the GLPs are predicted to have a high content of beta-pleated sheet. Seven conserved regions of beta-sheet were found in each of the GLP proteins along with alpha-helices located at both N- and C-termini. These same structural elements are also conserved in wheat germin. With one exception, all GLP family members contain at least one N-glycosylation site. All of these sites are conserved in an unstructured loop between beta-1 and beta-2. Genes for two of these GLPs were identified in genomic sequences previously deposited in the GenBank. The GLP3b gene is physically linked to the polyubiquitin 4 gene. The 3' end of the GLP3b mRNA is only 0.5 kb from the ubq4 start of transcription. Analysis of the GLP3b promoter shows the presence of a single putative auxin response sequence located at -124 to -111 upstream from the 5' end of the GLP3b mRNA. The GLP9 gene was identified in an Arabidopsis contig from Chromosome 4. PMID- 9869401 TI - Molecular cloning and characterization of the Amylose-Extender gene encoding starch branching enzyme IIB in maize. AB - The amylose-extender (Ae) gene encoding starch-branching enzyme IIb (SBEIIb) in maize is predominantly expressed in endosperm and embryos during kernel development. A maize genomic DNA fragment (-2964 to +20,485) containing the Ae gene was isolated and sequenced. The maize Ae mRNA is derived from 22 exons distributed over 16,914 bp. Twenty-one introns, differing in length from 76 bp to 4020 bp, all have conserved junction sequences (GT..AG). Sequence analysis of the 5'- and 3'-flanking regions revealed a consensus TATA-box sequence located 28 bp upstream of the transcription initiation site as determined by primer extension analysis, and a putative polyadenylation signal observed 29 bp upstream of the polyadenylation site based on cDNA sequence. Genomic Southern blot analysis suggests that a single Ae gene is present in the maize genome. Promoter activity was confirmed by testing a transcriptional fusion of the Ae 5'-flanking region between -2964 and +100 to a luciferase reporter gene in a transient expression assay using maize endosperm suspension cultured cells. 5' deletion analysis revealed that the 111 bp region from -160 to -50 is essential for high-level promoter activity. PMID- 9869402 TI - Reduced toxicity and broad spectrum resistance to viral and fungal infection in transgenic plants expressing pokeweed antiviral protein II. AB - Pokeweed antiviral protein II (PAPII), a 30 kDa protein isolated from leaves of Phytolacca americana, inhibits translation by catalytically removing a specific adenine residue from the large rRNA of the 60S subunit of eukaryotic ribosomes. The protein sequence of PAPII shows only 41% identity to PAP and PAP-S, two other antiviral proteins isolated from pokeweed. We isolated a cDNA corresponding to PAPII and introduced it into tobacco plants. PAPII expressed in transgenic tobacco was correctly processed to the mature form as in pokeweed and accumulated to at least 10-fold higher levels than wild-type PAP. We had previously observed a significant decrease in transformation frequency with PAP and recovered only two transgenic lines expressing 1-2 ng per mg protein. In contrast, eight different transgenic lines expressing up to 250 ng/mg PAPII were recovered, indicating that PAPII is less toxic than PAP. Two symptomless transgenic lines expressing PAPII were resistant to tobacco mosaic virus, potato virus X and the fungal pathogen Rhizoctonia solani. The level of viral and fungal resistance observed correlated well with the amount of PAPII protein accumulated. Pathogenesis-related protein PR1 was constitutively expressed in transgenic lines expressing PAPII. Although PR1 was constitutively expressed, no increase in salicylic acid levels was detected, indicating that PAPII may elicit a salicylic acid-independent signal transduction pathway. PMID- 9869403 TI - The ECS1 gene of Arabidopsis encodes a plant cell wall-associated protein and is potentially linked to a locus influencing resistance to Xanthomonas campestris. AB - The interaction between Arabidopsis and Xanthomonas campestris pv. campestris (Xcc) provides a useful model system to identify components that are involved in incompatible interactions of this phytopathogen and cruciferous plants. We have previously described a new gene from Arabidopsis thaliana, ECS1 (formerly CXC750), which showed an ecotype-specific expression pattern, although homologous sequences are present in the genomes of all ecotypes tested. Interestingly, ECS1 mRNA transcripts were only detected in ecotypes which showed the resistant phenotype against Xcc race 750. Subsequent genetic studies with F3 progeny of a cross between a Xcc750 resistant ecotype (Col-0) and a Xcc750 sensitive ecotype (Oy-0) revealed segregation of the ECS1RNA-phenotype away from the Xcc750 susceptible phenotype in one out of 16 F3 families. In addition, transformation of Xcc750 sensitive plants that did not express ECS1, with a constitutively transcribed ECS1 gene, did not lead to the resistant phenotype in the transgenic plants. From these results we conclude that ECS1 is not a Xcc750 resistance gene, but the genetic data indicate that ECS1 is linked to a locus influencing resistance to Xcc750. ECS1 was localized to YACs 3H12 and 12F3 of the CIC library, which map to chromosome 1. Subcellular localization of the gene product indicated that the ECS1 protein is associated with the plant cell wall. Further molecular investigation of 10 ecotypes revealed the utility of ECS1 as a new marker for chromosome 1, which is detectable by RFLP, PCR or expression analysis. PMID- 9869404 TI - Differential regulation of genes encoding 1-aminocyclopropane-1-carboxylate (ACC) synthase in etiolated pea seedlings: effects of indole-3-acetic acid, wounding, and ethylene. AB - Treatment of 5- to 6-day-old etiolated pea (Pisum sativum L.) seedlings with indole-3-acetic acid (IAA) induced within 15 min an increase in the transcript levels of two genes encoding 1-aminocyclopropane-1-carboxylate (ACC) synthase, Ps ACS1 and Ps-ACS2. Simultaneous treatment with ethylene inhibited this increase and also caused a decrease in ACC synthase enzyme activity as compared to that of seedlings treated with IAA alone. These results indicate that ethylene inhibits its own biosynthesis by decreasing ACC synthase transcript levels via a negative feedback loop. Wounding of pea stems had no effect on the expression of Ps-ACS1, but led within 10 min to an increase in the mRNA levels of Ps-ACS2. This increase was also inhibited by ethylene. The wound signal was transmitted over a distance of at least 4 cm through the stem with no delay in induction or response intensity. The rapid transmission of the wound response is consistent with the possibility that a hydraulic or electric signal is responsible for the spread of the wound response. PMID- 9869405 TI - Isolation and characterization of a rice homebox gene, OSH15. AB - In many eukaryotic organisms including plants, homeobox genes are thought to be master regulators that establish the cellular or regional identities and specify the fundamental body plan. We isolated and characterized a cDNA designated OSH15 (Oryza sativa homeobox 15) that encodes a KNOTTED-type homeodomain protein. Transgenic tobacco plants overexpressing the OSH15 cDNA showed a dramatically altered morphological phenotype caused by disturbance of specific aspects of tobacco development, thereby indicating the involvement of OSH15 in plant development. We analyzed the in situ mRNA localization of OSH15 through the whole plant life cycle, comparing the expression pattern with that of another rice homeobox gene, OSH1. In early embryogenesis, both genes were expressed as the same pattern at a region where the shoot apical meristem would develop later. In late embryogenesis, the expression pattern of the two genes became different. Whereas the expression of OSH1 continued within the shoot apical meristem, OSH15 expression within the shoot apical meristem ceased but became observable in a ring shaped pattern at the boundaries of some embryonic organs. This pattern of expression was similar to that observed around vegetative or reproductive shoots, or the floral meristem in mature plants. RNA in situ localization data suggest that OSH15 may play roles in the shoot organization during early embryogenesis and thereafter, OSH15 may be involved in morphogenetic events around the shoot apical meristem. PMID- 9869406 TI - Alfalfa (Medicago sativa L.) resistance to the root-lesion nematode, Pratylenchus penetrans: defense-response gene mRNA and isoflavonoid phytoalexin levels in roots. AB - Alfalfa (Medicago sativa) varieties with antibiosis-based resistance to the root lesion nematode (Pratylenchus penetrans), a migratory endoparasite of many crops, have been developed by recurrent selection. Individual plants from these varieties that support significantly lower nematode reproduction were identified for molecular and biochemical characterization of defense responses. Before nematode infection, RNA blot analysis revealed 1.3-1.8-fold higher phenylpropanoid pathway mRNA levels in roots of three resistant plants as compared to three susceptible alfalfa plants. The mRNAs encoded the first enzyme in the pathway (phenylalanine ammonia-lyase), the first in the pathway branch for flavonoid biosynthesis (chalcone synthase), a key enzyme in medicarpin biosynthesis (isoflavone reductase) and a key enzyme in the pathway branch for biosynthesis of lignin cell wall precursors (caffeic acid O-methyltransferase). After nematode infection, the mRNAs declined over 48 h in resistant roots but rose in susceptible plants during the first 12 h after-infection and then declined. Acidic beta-1,3-glucanase mRNA levels were initially similar in both root types but accumulated more rapidly in resistant than in susceptible roots after nematode infection. Levels of a class I chitinase mRNA were similar in both root types. Histone H3.2 mRNA levels, initially 1.3-fold higher in resistant roots, declined over 6-12 h to levels found in susceptible roots and remained stable in both root types thereafter. Defense-response gene transcripts in roots of nematode-resistant and susceptible alfalfa plants thus differed both constitutively and in inductive responses to nematode infection. HPLC analysis of isoflavonoid-derived metabolites of the phenylpropanoid pathway revealed similar total constitutive levels, but varying relative proportions and types, in roots of the resistant and susceptible plants. Nematode infection had no effect on isoflavonoid levels. Constitutive levels of the phytoalexin medicarpin were highest in roots of the two most resistant plants. Medicarpin inhibited motility of P. penetrans in vitro. PMID- 9869407 TI - Metabolic engineering of rice leading to biosynthesis of glycinebetaine and tolerance to salt and cold. AB - Genetically engineered rice (Oryza sativa L.) with the ability to synthesize glycinebetaine was established by introducing the codA gene for choline oxidase from the soil bacterium Arthrobacter globiformis. Levels of glycinebetaine were as high as 1 and 5 micromol per gram fresh weight of leaves in two types of transgenic plant in which choline oxidase was targeted to the chloroplasts (ChlCOD plants) and to the cytosol (CytCOD plants), respectively. Although treatment with 0.15 M NaCl [corrected] inhibited the growth of both wild-type and transgenic plants, the transgenic plants began to grow again at the normal rate after a significantly less time than the wild-type plants after elimination of the salt stress. Inactivation of photosynthesis, used as a measure of cellular damage, indicated that ChlCOD plants were more tolerant than CytCOD plants to photoinhibition under salt stress and low-temperature stress. These results indicated that the subcellular compartmentalization of the biosynthesis of glycinebetaine was a critical element in the efficient enhancement of tolerance to stress in the engineered plants. PMID- 9869408 TI - Identification of class B and class C floral organ identity genes from rice plants. AB - The functions of two rice MADS-box genes were studied by the loss-of-function approach. The first gene, OsMADS4, shows a significant homology to members in the PISTILLATA (PI) family, which is required to specify petal and stamen identity. The second gene, OsMADS3, is highly homologous to the members in the AGAMOUS (AG) family that is essential for the normal development of the internal two whorls, the stamen and carpel, of the flower. These two rice MADS box cDNA clones were connected to the maize ubiquitin promoter in an antisense orientation and the fusion molecules were introduced to rice plants by the Agrobacterium-mediated transformation method. Transgenic plants expressing antisense OsMADS4 displayed alterations of the second and third whorls. The second-whorl lodicules, which are equivalent to the petals of dicot plants in grasses, were altered into palea/lemma-like organs, and the third whorl stamens were changed to carpel-like organs. Loss-of-function analysis of OsMADS3 showed alterations in the third and fourth whorls. In the third whorl, the filaments of the transgenic plants were changed into thick and fleshy bodies, similar to lodicules. Rather than making a carpel, the fourth whorl produced several abnormal flowers. These phenotypes are similar to those of the agamous and plena mutants in Arabidopsis and Antirrhinum, respectively. These results suggest that OsMADS4 belongs to the class B gene family and OsMADS3 belongs to the class C gene family of floral organ identity determination. PMID- 9869409 TI - Macromolecular organization and genetic mapping of a rapidly evolving chromosome specific tandem repeat family (B77) in cotton (Gossypium). AB - Isolation and characterization of the most prominent repetitive element families in the genome of tetraploid cotton (Gossypium barbadense L; [39]) revealed a small subset of families that showed very different properties in tetraploids than in their diploid progenitors, separated by 1-2 million years. One element, B77, was characterized in detail, and compared to the well-conserved 5S and 45S rRNA genes. The 572 bp B77 repeat was found to be concentrated in several discontinuous tandem arrays confined to a single 550 kb SalI fragment in tetraploid cotton. Genetic mapping based on the absence of the pentameric 'rung' in the G. barbadense 'ladder' showed that B77 maps to a D-subgenome chromosome. In situ hybridization supports the contention that the array is confined largely to a single chromosomal site in the D-subgenome. The B77 repeat has undergone a substantial increase in copy number since formation of tetraploid cotton from its diploid relatives. RFLPs observed among tetraploid cotton species suggest that amplification and/or rearrangement of the repeat may have continued after divergence of the five tetraploid cotton species. B77 contains many short direct repeats and shares significant DNA sequence homology with a Nicotiana alata retrotransposon Tna1-2 integrase motif. The recent amplification of B77 on linkage group D04 suggests that the D-subgenome of tetraploid cotton may be subject to different evolutionary constraints than the D-genome diploid chromosomes, which exhibit few genome-specific elements. Further, the abundance of B77 in G. gossypioides supports independent evidence that it may be the closest extant relative of the D-genome ancestor of cotton. PMID- 9869410 TI - Physical mapping of unique nucleotide sequences on identified rice chromosomes. AB - A physical mapping method for unique nucleotide sequences on specific chromosomal regions was developed combining objective chromosome identification and highly sensitive fluorescence in situ hybridisation (FISH). Four unique nucleotide sequences cloned from rice genomic DNAs, varying in size from 1.3 to 400 kb, were mapped on a rice chromosome map. A yeast artificial chromosome (YAC) clone with a 399 kb insert of rice genomic DNA was localised at the distal end of the long arm of rice chromosome (1q2.1) and a bacterial artificial chromosome (BAC) clone (180 kb) containing the rice leaf blast-resistant gene (Pi-b) was shown to occur at the distal end of the long arm of chromosome 2 (2q2.1). A cosmid (35 kb) with the resistance gene (Xa-21) against bacterial leaf blight was mapped on the interstitial region of the long arm on chromosome 11 (11q1.3). Furthermore a single RFLP marker, 1.29 kb in size, was mapped successfully to the distal region of the long arm of rice chromosome 4 (4q2.1). For precise localisation of the nucleotide sequences within the chromosome region, image analyses were effective. The BAC clone was localised to the specific region, 2q2.1:96.16, by image analysis. The result was compared with the known location of the BAC clone on the genetic map and the consistency was confirmed. The effectiveness and reliability in physically mapping nucleotide sequences on small plant chromosomes achieved by the FISH method using a variety of probes was unequivocally demonstrated. PMID- 9869411 TI - Gibberellin and abscisic acid regulate GAST1 expression at the level of transcription. AB - Both gibberellic acid (GA3) and abscisic acid (ABA) regulate the expression of the GAST1 gene of tomato. Treatment with GA3 increases the abundance of GAST1 RNA while treatment with ABA blocks this effect. In this study, the effects of GA3 and ABA on the rate of transcription of the GAST1 gene and the stability of GAST1 RNA were examined. Nuclear run-on analyses detected an increase in transcription of the GAST1 gene 1 h after GA3 treatment with transcription increasing to a maximum at 9 h after treatment. The half-life of GAST1 RNA in GA3-treated leaves was similar to that in control leaves. In addition, the extent of overexpression of GAST1 RNA in transgenic tomato plants containing the CaMV 35S promoter driving the expression of the GAST1 transcribed region was largely unaffected by GA3. These results suggest that GA3 stimulates the expression of the GAST1 gene by acting only at the level of transcription. ABA treatment dramatically reduced the abundance of GAST1 RNA in gib1 shoots through an effect at the level of transcription and did not appear to affect the stability of this RNA. Midcourse ABA addition to the GA3-incubated shoots reversed the GA3-mediated increase in the transcription of GAST1 gene within 15 min. Transgenic plants that either overexpressed or underexpressed GAST1 RNA exhibited no phenotypic differences from wild type. PMID- 9869412 TI - Wild and cultivated barleys show differences in the expression pattern of a cold regulated gene family under different light and temperature conditions. AB - Cold acclimation in plants involves the expression of many genes and gene families. The present study reports the expression analysis of three members of the blt14 gene family in barley. Gene-specific antisense oligonucleotides were used as probes in northern experiments so as to follow independently the expression of individual members of the gene family. Each clone revealed different accumulation kinetics when a spring and a winter cultivar were compared, suggesting that the different regulatory mechanisms leading to mRNA accumulation of an individual member of the blt14 gene family are genotype dependent. In a collection of Hordeum spontaneum genotypes both qualitative and quantitative polymorphisms were found for the accumulation of blt14-related mRNAs, although no clear relationships were found between blt14 expression and frost resistance. The accumulation of the blt14-related mRNAs was also modulated by light and by the albino mutation a(n). The effects of light on the accumulation of the transcripts corresponding to the blt14 gene family were evaluated by comparing etiolated and green plants. Etiolated plants accumulate the blt14-related mRNAs at a detectable level already at 22 degrees C. When the same plants are exposed to cold in absence of light an increased mRNA accumulation above the level present in green cold-treated plants can be detected. On the contrary, etiolated plants showed a reduced blt14 accumulation when exposed to cold in the presence of light. Cold-induced expression of the blt14 gene family was strongly reduced in plants carrying the albino mutation a(n). This mutant showed a defective molecular response to cold even when probed with a cDNA coding for LEA type protein (paf93). The albino mutant a(n) was not able to harden when exposed to low temperature providing a direct evidence of the relationship between expression of cold-regulated (COR) genes and the development of cold hardening. Failure of cold acclimation in the mutant cannot be merely ascribed to the absence of photosynthetic activity, since etiolated wild-type plants accumulated COR mRNAs and improved frost resistance when exposed to cold. PMID- 9869413 TI - The promoter of the plant defensin gene PDF1.2 from Arabidopsis is systemically activated by fungal pathogens and responds to methyl jasmonate but not to salicylic acid. AB - The plant defensin PDF1.2 has previously been shown to accumulate systemically via a salicylic acid-independent pathway in leaves of Arabidopsis upon challenge by fungal pathogens. To further investigate the signalling and transcriptional processes underlying plant defensin induction, a DNA fragment containing 1184 bp and 1232 bp upstream of the transcriptional and translational start sites, respectively, was cloned by inverse PCR. To test for promoter activity this DNA fragment was linked to the beta-glucuronidase (GUS)-encoding region of the UidA gene as a translational fusion and introduced into Arabidopsis ecotype C-24. Challenge of the transgenic plants with the fungal pathogens Alternaria brassicicola and Botrytis cinerea resulted in both local and systemic induction of the reporter gene. Wounding of the transgenic plants had no effect on GUS activity. Treatment of the transgenic plants with either jasmonates or the active oxygen generating compound paraquat strongly induced the reporter gene. In contrast, neither salicylate nor its functional analogues 2,6 dichloroisonicotinic acid and 1,2,3-benzothiodiazole-7-carbothioic acid S-methyl ester resulted in reporter gene induction. These results are consistent with the existence of a salicylic acid-independent signalling pathway, possibly involving jasmonates as regulators, that is triggered by pathogen challenge but not by wounding. The transgenic plants containing the PDF1.2-based promoter-reporter construct will provide useful tools for future genetic dissection of this novel systemic signalling pathway. PMID- 9869414 TI - Visualization of the Brassica self-incompatibility S-locus on identified oilseed rape chromosomes. AB - Seventy years after Karpechenko [15] first reported the accurate chromosome number of oilseed rape (Brassica napus L., 2n=38), we have developed a quantitative chromosome map of rape using computer imaging technology. The capacity to identify individual rape chromosomes will facilitate a wide range of genetic studies. Here we demonstrate the use of imaging methods in combination with fluorescence in situ hybridization to localize, on identified chromosomes, the single copy S-locus glycoprotein and S-locus-related genes involved in the self-incompatibility system of Brassica. These techniques have a broader application in plant genome research involving the mapping of single-copy genes and markers, irrespective of the plant species. PMID- 9869415 TI - Desiccation- and abscisic acid-responsive genes encoding major intrinsic proteins (MIPs) from the resurrection plant Craterostigma plantagineum. AB - Major intrinsic proteins (MIPs) are a family of channel proteins that are mainly represented by aquaporins in plants. These are divided into TIPs (tonoplast intrinsic proteins) and PIPs (plasma membrane intrinsic proteins) according to their subcellular localization. Homologues to PIPs and TIPs were isolated from the desiccation-tolerant resurrection plant Craterostigma plantagineum by two approaches: firstly, a cDNA library constructed from RNA of dehydrated C. plantagineum leaves was screened with an Arabidopsis thaliana Ath-PIP1b cDNA probe and, secondly, a cDNA library was screened differentially to isolate early drought-induced transcripts. According to sequence homologies the isolated cDNA clones were grouped as follows: Cp-PIPa, Cp-PIPb, Cp-PIPc and Cp-TIP. Cp-PIPa, Cp PIPc and Cp-TIP transcript accumulation was regulated by dehydration and abscisic acid (ABA). Within the Cp-PIPa group transcripts were regulated either by drought only or by drought and ABA, indicating that ABA-dependent and -independent signal transduction pathways lead to Cp-PIPa expression. Comparison of Cp-PIPa expression in detached leaves and in whole plants suggested the involvement of a signal transmitted in the whole plant in response to drought. Cp-PIPb transcript levels were constitutive in all organs tested. Antibodies raised against a Cp PIPA protein recognized a polypeptide with an apparent molecular mass of 28 kDa. Using these antibodies it was shown that both Cp-PIPA and Cp-PIPB proteins were localized to the plasma membrane. The role of different members of the MIP group in the dehydration response is discussed. PMID- 9869416 TI - Differential induction by methyl jasmonate of genes encoding ornithine decarboxylase and other enzymes involved in nicotine biosynthesis in tobacco cell cultures. AB - A cDNA of tobacco BY-2 cells corresponding to an mRNA species which was rapidly induced by methyl jasmonate (MeJA) in the presence of cycloheximide (CHX) was found to encode ornithine decarboxylase (ODC). Another cDNA from a MeJA-inducible mRNA encoded S-adenosylmethionine synthase (SAMS). Although these enzymes could be involved in the biosynthesis of polyamines, the level of putrescine, a reaction product of ODC, increased slowly and while the levels of spermidine and spermine did not change following treatment of cells with MeJA. However, N methylputrescine, which is a precursor of pyrrolidine ring of nicotine, started to increase shortly after MeJA-treatment of cells and the production of nicotine occured thereafter. The levels of mRNA for arginine decarboxylase (ADC), an alternative enzyme for putrescine synthesis, and that for S-adenosylmethionine decarboxylase (SAMDC), required for polyamine synthesis, were not affected by MeJA. In addition to mRNAs for ODC and SAMS, mRNA for putrescine N methyltransferase (PMT) was also induced by MeJA. Unlike the MeJA-induction of ODC mRNA, MeJA-induction of SAMS and PMT mRNAs were blocked by CHX. The level of ODC mRNA declined after 1 to 4 h following MeJA treatment, while the levels of mRNAs for SAMS and PMT continued to increase. Auxin significantly reduced the MeJA-inducible accumulation of mRNAs for ODC, SAMS and PMT. These results indicate that MeJA sequentially induces expression of a series of genes involved in nicotine biosynthesis by multiple regulatory mechanisms. PMID- 9869417 TI - Longevity of 5-azacytidine-mediated gene expression and re-establishment of silencing in transgenic rice. AB - Epigenetic silencing of a bialaphos resistance (bar) gene in R1 progeny of a transgenic rice line was found to be meiotically stable since selfed (R2) progeny were also susceptible and the bar locus highly methylated. A high proportion of R2 seedlings germinated in the presence of 5-azacytidine (AzaC) were herbicide resistant and also contained at least one unmethylated copy of the bar gene, further establishing the relationship between silencing and methylation. Restored bar gene expression was typically maintained for 20-50 days, but eventual methylation and silencing of the bar locus underscores the ability of the recipient genome to recognize and inactivate intrusive DNA. PMID- 9869418 TI - Alfin1, a novel zinc-finger protein in alfalfa roots that binds to promoter elements in the salt-inducible MsPRP2 gene. AB - Alfin1 cDNA, obtained by differential screening of a poly(A)+ library from salt tolerant alfalfa cells, encodes a novel protein with a Cys4 and His/Cys3 putative zinc-binding domain that suggests a possible role for this protein in transcriptional regulation. We have expressed the cDNA in Escherichia coli and show that the recombinant Alfin1 protein binds DNA in a sequence-specific manner. The DNA recognition sequence was determined from individual clones isolated after four rounds of random oligonucleotide selection in gel retardation assays, coupled with PCR amplification of the selected sequences. The consensus binding site for Alfin1 is shown to contain two to five G-rich triplets with the conserved core of GNGGTG or GTGGNG in clones showing high-efficiency binding. DNA binding of the recombinant Alfin1 was inhibited by EDTA. Alfin1 mRNA was found predominantly in alfalfa roots. Growth of salt-sensitive Medicago sativa L on 171 mM NaCl led to a slight decrease in Alfin1 mRNA, while the salt-tolerant plants showed no decrease in Alfin1 mRNA levels. Interestingly, recombinant Alfin1 binds efficiently to three fragments of the MsPRP2 promoter, each containing consensus sequences identified by the random oligonucleotide selection. Since MsPRP2 transcripts were shown to be root-specific and accumulated in alfalfa roots in a salt-inducible manner, Alfin1 may play a role in the regulated expression of MsPRP2 in alfalfa roots and contribute to salt tolerance in these plants. PMID- 9869419 TI - Characterization of tomato PHYB1 and identification of molecular defects in four mutant alleles. AB - The structure of the gene encoding the apoprotein of phytochrome B (PHYB1) in tomato has been determined from genomic and cDNA sequences. In contrast to PHYA, PHYB1 lacks an intron upstream of the first ATG. A single transcription start site was found by 5' RACE at -116. Tomato PHYB1 spans 7 kb starting from the first ATG. The coding region is organized into four exons as for other angiosperm PHY. The deduced apoprotein consists of 1131 amino acids, with a molecular mass of 125.4 kDa. Tomato phytochrome B1 shares 78% and 74% identity with Arabidopsis phytochromes B and D, respectively. Along with the normally spliced full-length transcripts, sequences of reverse transcriptase-PCR clones revealed five types of alternative transcripts. Each type of alternative transcript was missing a considerable part of the coding region, including the chromophore-binding site. The four putative PHYB1 mutants in tomato, which are temporarily red-light insensitive (tri), were each confirmed to have a mutation in PHYB1. Each mutation arose from a different, single-base substitution. Allele tri1 is presumably a null because the mutation introduces a stop at codon 92. In tri3, val-238 is replaced by Phe. The importance of this valine residue is evidenced by the fact that the tri3 phenotype is as strong as that of tri1. Alleles tri2 and tri4 encode proteins truncated at their C-termini. The former lacks either 170 or 438 amino acids, depending upon which of two types of splicing occurs during transcript maturation, while the latter lacks 225. PMID- 9869421 TI - Jasmonic acid stimulates the expression of nod genes in Rhizobium. AB - Jasmonates and salicylic acid are considered to be signal molecules that induce a variety of plant genes involved in wound or defence response, as well as affecting nos promoter activity. In this paper we examined whether these chemicals could also affect nod genes from isogenic rhizobia strains. Isogenic strains contain the Rhizobium leguminosarum nodA promoter fused to the lacZ gene of Escherichia coli and differ only in the source of the regulatory nodD gene. Naringenin, jasmonic acid and methyl jasmonate induced expression of nod genes in strain RBL1284 and salicylic acid showed no activity alone or when used in combination with other compounds; addition of naringenin + jasmonic acid produced a synergistic effect. Results obtained with strain RBL5284 were similar to those for RBL1284 albeit the combination of naringenin with the other compounds markedly inhibited nod gene expression. Whereas RBL5283 responded to naringenin with a strong induction, jasmonic acid, methyl jasmonate or salicylic acid showed no significant responses. The inhibitory effect of salicylic acid on nod gene expression indicates that the induction mechanism of jasmonic acid, methyl jasmonate, N-propyldihydrojasmonate and naringenin is probably different from that of salicylic acid. PMID- 9869420 TI - CaMADS1, a MADS box gene expressed in the carpel of hazelnut. AB - Hazelnut (Corylus avellana L.) is a species of economic interest that shows a peculiar floral biology. Unlike most of the angiosperms, which produce ovules during floral development such that they are ready for pollen at anthesis, hazelnut ovary development is delayed and triggered by compatible pollination. In order to elucidate the mechanisms regulating this unusual process and the role of the MADS box genes in ovary development, a cDNA library from pollinated styles of hazelnut was screened with a mixture of MADS box genes from different plant species. CaMADS1 (Corylus avellana MADS box), a floral-specific MADS box gene, was isolated, and characterized as belonging to the sub-family of the AGAMOUS genes. Northern blot, RT-PCR analyses and in situ hybridization experiments show a precise correlation between ovary development and CaMADS1 expression, indicating a role of this MADS box gene in the processes of floral organogenesis. PMID- 9869422 TI - Expression of an anther-specific chalcone synthase-like gene is correlated with uninucleate microspore development in Nicotiana sylvestris. AB - Two cDNA clones, specifically expressed in Nicotiana sylvestris anthers during uninucleate microspore development, were isolated using a subtractive hybridization approach. Sequence analysis showed that one of them, NSCHSLK, displayed a high level of similarity to several anther-specific chalcone synthase like (CHSLK) proteins and an ORF from chromosome I of Arabidopsis thaliana. A lower, but significant, similarity to chalcone synthases and closely related enzymes (CHSRE) was also detected. The structure of the nschslk gene was found to be typical of the chalcone (chs)/stilbene (sts) synthase family. Expression of NSCHSLK mRNA was confined to microspores and tapetal cells. UV-irradiation or infection with Phytophthora parasitica var. nicotianae of transgenic Nicotiana benthamiana plants carrying a chimeric nschslk/GUS gene indicated that the nschslk promoter exhibits the same anther-specific, developmentally regulated expression pattern. Comparison of CHSRE and CHSLK polypeptide sequences revealed some important similarities and differences between the two groups. The data presented in this study, suggest that the anther-specific chslk genes represent a separate sub-family of plant polyketide synthases related to chs/sts in terms of gene structure, polypeptide sequence and the possible catalytic mechanism, but differing in substrate/product specificity. The putative role of CHSLK enzymes in anther development and particularly in exine synthesis is discussed. PMID- 9869423 TI - Characterization of a germin-like protein gene expressed in somatic and zygotic embryos of pine (Pinus caribaea Morelet). AB - Germin-like proteins (GLPs) ionically bound to the walls of preglobular somatic embryos of Pinus caribaea Morelet are markers of this early developmental stage. In order to reveal the physiological implications of such markers during early embryo development, we isolated a cDNA clone from somatic embryos predicted to encode a protein with sequence similarity to GLPs. PcGER1 has an open reading frame corresponding to a 220 amino acid polypeptide with a putative N glycosylation site on Asn-69. The presence of a 24 amino acid putative signal peptide supports the hypothesis of an apoplastic location. The N-terminal 20 amino acid sequence of the predicted mature protein is identical to the amino terminal sequence of GP111, one of the extracellular pine GLPs previously identified. Southern blot hybridizations indicate that PcGER1 is probably unique in the pine genome. Transcripts homologous to PcGER1 are abundant in all embryogenic lines, absent from nonembryogenic lines, and present in quiescent zygotic embryos but not in the female gametophyte, the haploid storage tissue of conifers. Their abundance sharply decreases during germination. Isolation of gf 0.8, a genomic fragment identical to PcGER1 cDNA sequence, confirms that no introns disrupt the coding region as it has been already described for wheat gf 2.8 and gf-3.8 genomic clones. Recombinant PcGER1, produced in Escherichia coli, is recognized by antibodies raised against the GP111 N-terminal nonapeptide and the unglycosylated wheat germin monomer. The implications of GLPs in pine embryogenesis are discussed. PMID- 9869424 TI - Substitution of Ala-251 of the D1 reaction centre polypeptide with a charged residue results in impaired function of photosystem II. AB - Ala-251 in the membrane-parallel helix in the D-E loop of the D1 polypeptide close to the Q(B) pocket of photosystem II (PS II), was mutated to aspartate (D), lysine (K), leucine (L) or serine (S) in Synechocystis 6803. O2 evolution rates (H2O-->DCBQ; 2,6-dichloro-p-benzoquinone) of A251D, A251L and A251S were lower, being 38, 16, 62 and 70%, respectively, of that of the control, and there was an even more drastic impairment of O2 evolution when measured from H2O to DMBQ (2,5 dimethyl-p-benzoquinone), demonstrating modifications in the Q(B) pocket. However, in all other mutants but A251K, the Q(B) function could sustain O2 evolution at a level high enough to support photosynthetic growth. The mutant A251S, carrying a substitution of alanine for a chemically quite similar residue serine, was less severely affected. Substitution by a positively charged residue drastically delayed chlorophyll a fluorescence relaxation in the non photosynthetic strain A251K, implying strong impairment of Q(A)-to-Q(B) electron transfer. Delay of fluorescence relaxation was clear in A251D as well, carrying a substitution of alanine for a negatively charged residue. The effects of the substitutions of A251 demonstrate the importance of this residue of the D1 polypeptide in the conformation of the acceptor side of PS II and, accordingly, the effect on the acceptor-side function of PS II was very clear. Nevertheless, the tolerance of PS II activity to high-light-induced photoinhibition in vivo and the subsequent D1 degradation were not much impaired in any of the photosynthetic mutant strains as compared to the control. PMID- 9869425 TI - Anthranilate N-hydroxycinnamoyl/benzoyltransferase gene from carnation: rapid elicitation of transcription and promoter analysis. AB - Hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT) catalyses the committed reaction of phytoalexin biosynthesis in carnation (Dianthus caryophyllus L.). Three HCBT cDNAs were cloned previously from suspension-cultured carnation cells that had been induced with fungal elicitor. A rapid, transient induction of hcbt transcripts upon elicitation, reaching maximal abundances within about 0.5 h and returning to basal levels within 4 h, suggested the involvement of unusual cis elements. A DNA fragment of 3.8 kb, spanning the hcbt2 gene with the 5'-flanking region of roughly 1.8 kb, was cloned from carnation plants. The gene encodes one long open reading frame lacking introns. The DNA sequence revealed a conserved TATA box, three elicitor response elements (EREs) and a 9 bp direct repeat as well as an interrupted direct repeat of 11 bp in the TATA distal region. EMSA revealed the binding of elicitor-inducible nuclear factors to the promoter region from -377 to -326 spanning two of the EREs, and their functional relevance was confirmed by transient expression assays of hcbt2 promoter-GUS reporter gene constructs in parsley protoplasts. Furthermore, an oligo(A) segment was present immediately preceding the start of translation (+140 to +150). Transient expression analysis demonstrated that the sequence upstream to -1157 at least is required in context with the 5'-UTR, particularly including the poly(A) segment, for strong expression and full elicitor induction of the hcbt2 gene. The results suggested that several sequence motifs scattered over a wide range of the 5'-flanking region and into the exonic sequence are responsible for the full elicitor regulation of the hcbt2 gene. PMID- 9869426 TI - Identification of phosphoenolpyruvate carboxylase isoforms in leaf, stem and roots of the obligate CAM plant Vanilla planifolia Salib. (Orchidaceae): a physiological and molecular approach. AB - This study provides the first comparative analysis of phosphoenolpyruvate carboxylase isoforms (PEPc; EC 4.1.1.31) in an obligate crassulacean acid metabolism (CAM) plant, Vanilla planifolia Salisb. (Orchidaceae). Nocturnal CO2 fixation and malate accumulation by the leaves and the green stem show that these organs perform CAM. The chloroplast-containing aerial roots, however, exhibit C3 photosynthesis. The catalytic activity of PEPc was highest in the leaves compared with the stem and aerial roots. The Km (PEP) and Ki (malate) were similar in the PEPc extracted from leaf and aerial roots, and significant higher in stem. cDNA was obtained from those tissues and also from the soil-grown roots, and various cDNA clones were detected and amplified by means of RT-PCR and RACE-PCR. The amino-acid sequences of the PEPc isoforms deduced from the cDNA showed a great degree of homology, and Southern blot analysis suggests that the encoding genes form a small multigene family of at least two members. One PEPc isoform (PpcV1) is assumed to be related to CAM because, as shown by northern blot analysis, it is mainly expressed in the CAM-performing organs, i.e. in the leaves and the stem. A further isoform (PpcV2) was identified in the soil-grown roots and aerial roots, but northern blots show that to some extent PpcV2 is also expressed in the leaf and the stem tissues. Thus, it is assumed that PpcV2 encodes the housekeeping isoform of PEPc. Altogether, the present study provides support in favour of the view that isoforms of PEPc are related to specific functions. PMID- 9869427 TI - Cloning of genes by mRNA differential display induced during the hypersensitive reaction of soybean after inoculation with Pseudomonas syringae pv. glycinea. AB - Soybean (Glycine max [L.] Merr.) cell suspension cultures (cv. Williams 82) inoculated with the pathogenic bacteria Pseudomonas syringae pv. glycinea respond with a hypersensitive reaction (HR) when the bacteria express the avirulence gene avrA. A mRNA differential display was established for this system to allow the identification of genes induced during the HR. Six PCR-fragments (DD1-DD6) from the differential display analysis were identified, which are induced during the HR. Database searches revealed that the fragment DD1 encodes chalcone isomerase and DD2 was identified as ubiquitin. The fragment DD3 shares significant homology to the signalling molecule 14-3-3. The partial DD4 product is homologous to the enhancer of rudimentary from Drosophila and an uncharacterized homologue of it from Arabidopsis. The fragment DD5 is similar to glucose-6-phosphate dehydrogenase which provides NADPH to the cell. The PCR-product DD6 seems to be a new leucine-rich-repeat disease resistance gene from soybean, which is significantly induced during the HR. All of the identified genes are clearly induced during a HR in infected plants of the same cultivar, indicating that results from the cell culture model system can be transferred to intact plants. These studies show that complex mRNA differential display is a powerful tool to identify new induced gene in plant-pathogen interactions. PMID- 9869428 TI - A chestnut seed cystatin differentially effective against cysteine proteinases from closely related pests. AB - Cystatin CsC, a cysteine proteinase inhibitor from chestnut (Castanea sativa) seeds, has been purified and characterized. Its full-length cDNA clone was isolated from an immature chestnut cotyledon library. The inhibitor was expressed in Escherichia coli and purified from bacterial extracts. Identity of both seed and recombinant cystatin was confirmed by matrix-assisted laser desorption/ionization mass spectrometry analysis, two-dimensional electrophoresis and N-terminal sequencing. CsC has a molecular mass of 11,275 Da and pI of 6.9. Its amino acid sequence includes all three motifs that are thought to be essential for inhibitory activity, and shows significant identity to other phytocystatins, especially that of cowpea (70%). Recombinant CsC inhibited papain (Ki 29 nM), ficin (Ki 65 nM), chymopapain (Ki 366 nM), and cathepsin B (Ki 473 nM). By contrast with most cystatins, it was also effective towards trypsin (Ki 3489 nM). CsC is active against digestive proteinases from the insect Tribolium castaneum and the mite Dermatophagoides farinae, two important agricultural pests. Its effects on the cysteine proteinase activity of two closely related mite species revealed the high specificity of the chestnut cystatin. PMID- 9869430 TI - TATA sequence requirements for the initiation of transcription for an RNA polymerase II in vitro transcription system from Nicotiana tabacum. AB - This is the first report to describe the detailed analysis of TATA sequence requirements for the initiation of transcription for RNA polymerase II in a plant in vitro transcription system. We examined the transcriptional activities associated with 24 promoters, including wild-type (TATATATA), and the 21 point and two multiple mutations in tobacco, Drosophila, and HeLa in vitro systems. Although the relative transcriptional activities varied among these promoters, all three systems gave similar results. These results suggest the functional conservation of TATA sequence requirements for in vitro transcription systems among plants, insects, and human. PMID- 9869429 TI - Sequential and structural homology between intracellular pathogenesis-related proteins and a group of latex proteins. AB - The intracellular pathogenesis-related proteins have been identified in a broad range of flowering plants. Some display quite different patterns of expression, in many cases unrelated to the pathogenic response. Nevertheless, these proteins are all very similar and in most cases share more than 35% sequence identity. In this report we investigate the significance of a rather weak similarity between the intracellular pathogenesis-related (IPR or PR-10) proteins and a group of proteins identified in the latex of opium poppy and in Arabidopsis, among others. A sequence analysis held together with the recently published three-dimensional structure of Bet v 1, an IPR protein from birch pollen, strongly suggests sequential and structural homology between the two protein families. PMID- 9869431 TI - A MADS box transcription factor of the AP1/AGL9 subfamily is also expressed in the seed coat of pea (Pisum sativum) during development. AB - As part of a programme to identify seed coat-specific transcripts in pea (Pisum sativum cv. Finale) using the differential display method, we identified and isolated a cDNA encoding a MADS box transcription factor. The predicted peptide contains 247 amino acids with the conserved N-terminal MADS box and is closely related to the AP1/AGL9 subfamily of the MADS box proteins. In addition to weak expression in petals, stamens and carpels, this MADS box mRNA is specifically expressed in the seed coat during seed development. PMID- 9869432 TI - Intergenic sequences of clustered tRNA genes: new type of genetic marker for phylogenetic studies, with application to the taxonomy of liverworts. AB - We have established a new type of genetic marker in Pellia liverworts that can be used to differentiate between closely related species within this genus. The Pellia intergenic sequences spanning the tandemly repeated nuclear tRNA(Leu) genes were amplified using the PCR technique and, in the case of Pellia neesiana, the tRNA(Leu) tandem region was cloned and sequenced. The length comparison of the intergenic regions allowed for the identification of a number of Pellia species. Additional experiments suggest that this marker can be used to differentiate other species of liverworts and mosses. PMID- 9869433 TI - Arterial grafting: techniques and conduits. AB - The superior long-term patency and survival of the internal thoracic artery in coronary artery bypass grafting, compared with saphenous vein, established the internal thoracic artery as the conduit of choice for myocardial revascularization. Use of the internal thoracic artery has expanded, and the possibility of similar performance by other arteries has motivated surgeons to investigate alternative arterial conduits (eg, the gastroepiploic artery, inferior epigastric artery, and radial artery). Although these grafts have become more technically feasible and have shown benefits, more follow-up data are needed to determine the long-term patency, freedom from arteriosclerosis, and efficacy of alternative conduits. PMID- 9869434 TI - The efficacy and safety of aprotinin use in cardiac surgery. AB - BACKGROUND: The serine protease inhibitor aprotinin has received much attention in cardiac surgical practice as a pharmacologic intervention to improve the hemostatic derangement associated with cardiopulmonary bypass. This review highlights the major studies undertaken to investigate the efficacy and safety of aprotinin use in both primary and repeat coronary artery bypass graft surgical procedures. METHODS: There have been at least 45 controlled studies in more than 7,000 patients in a variety of patient populations. These have ranged from primary coronary artery bypass graft and valve operations to complex reoperation procedures, including aortic arch reconstructions and thoracic organ transplantation. The recently completed International Multicenter Graft Patency Experience trial, the largest study to date, involved 870 patients at 13 international sites. The study examined the effects of aprotinin on graft patency, incidence of myocardial infarction, and blood loss in patients undergoing primary coronary artery bypass graft operations with cardiopulmonary bypass. RESULTS: Twenty-one studies in approximately 5,000 patients undergoing primary coronary artery bypass graft or valve operations reported 33% to 66% reduction in blood loss with full-dose aprotinin therapy; 15 of the same studies reported significant reductions in transfusion requirements, ranging from 31% to 85%. The recently completed International Multicenter Graft Patency Experience study observed a significant reduction in thoracic-drainage volume of 43% (p < 0.0001) and a 49% (p < 0.001) reduction in the requirement for allogeneic blood transfusions. Aprotinin did not affect the occurrence of definite myocardial infarction (aprotinin, 2.9% versus placebo, 3.8%) or mortality (aprotinin, 1.4% versus placebo, 1.6%). There was no observed difference in the patency of internal mammary artery bypass grafts from all study sites in aprotinin- versus placebo-treated patients (aprotinin, 98.2% versus placebo, 98.0%). CONCLUSIONS: Given the risks and costs associated with excessive bleeding and transfusions and the limited supply of banked blood, aprotinin represents an important and safe approach to blood conservation. PMID- 9869435 TI - Inflammatory response to cardiopulmonary bypass. AB - This article reviews the roles of the contact and complement systems and of neutrophils and monocytes in the inflammatory response to cardiopulmonary bypass and open heart operation. These blood proteins and cells, together with other blood elements, produce the vasoactive and cytotoxic substances and microemboli that cause the morbidity associated with cardiopulmonary bypass and open heart operation. PMID- 9869436 TI - Endothelial response to cardiopulmonary bypass surgery. AB - BACKGROUND: The vascular endothelium has been shown to actively participate in maintaining normal cardiovascular homeostasis by influencing the regulation of membrane permeability, lipid transport, vasomotor tone, coagulation, fibrinolysis, and inflammation. Endothelial cells are very responsive to a wide range of local and systemic stimuli that occur during cardiopulmonary bypass (CPB) operation. Major pathologic conditions result from impaired vascular function secondary to CPB, including vasospasm, coagulopathy, and widespread neutrophil adhesion secondary to a systemic inflammatory response. Additionally, more chronic responses to endothelial cell injury include the development of intimal hyperplasia and arteriosclerosis, both of which limit the long-term success of coronary artery bypass grafting. METHODS: Because of the increasingly recognized role of the endothelium in the maintenance of normal cardiovascular function, this article will review the normal structure and function of the endothelium, as well as the major pathologic conditions that result in response to CPB. RESULTS: Potential treatments to counteract endothelial cell dysfunction secondary to CPB are under active investigation. Strategies may be directed toward blocking single cytokines, integrins, or adhesion molecules involved in endothelial dysfunction or, alternatively, toward targeting a molecular event that governs the expression of these proinflammatory, procoagulant, and vasoactive genes. In our laboratory, we have used both strategies to study the pathologic response to CPB. We blocked neutrophil adhesion in subhuman primates with a monoclonal antibody. Alternatively, we targeted the transcriptional activation of multiple genes involved in the endothelial cell's response to CPB. CONCLUSIONS: Although both therapies help elucidate the multiple, redundant pathways involved in the pathologic response to CPB, it is through molecular biology that we are beginning to understand the mechanics of transcriptional control and translational expression that occurs in the endothelial cell in response to CPB. This knowledge will allow the development of therapies that inhibit not a single cytokine or adhesion molecule, but rather an array of substances that result in the endothelial cell's pathologic response to CPB. PMID- 9869437 TI - Central nervous system effects of cardiopulmonary bypass. AB - BACKGROUND: The spectrum of approaches to the issue of brain injury in cardiac surgical practice ranges from refusal to acknowledge that the problem exists to an overemphasis on cerebral risks that can unduly frighten patients. An appropriate approach to therapeutic and preventive strategies requires a fitting sense of proportion and an understanding of the mechanisms of cerebral injury. METHODS: This article reviews the incidence and severity of cerebral injury during cardiopulmonary bypass, the identification of high-risk patients, and the mechanisms of injury, including hypoperfusion, microemboli, and inflammatory response. It discusses the influences of alpha-stat and pH-stat strategies on cerebral blood flow during cardiopulmonary bypass; the use of retinal angiography to image the retinal circulation, thus providing a window on the cerebral microcirculation during bypass; magnetic resonance imaging evidence of an inflammatory response in the brain during bypass; and current efforts to gain better understanding of the molecular mechanisms involved in the inflammatory response. RESULTS: The current incidence of stroke during cardiopulmonary bypass is somewhat lower than in the 1980s but still remains a significant problem. Levels of cognitive impairment also are unacceptably high. Recognized predictors enable us to identify patients at particularly high risk of stroke. Hypertensive patients are particularly susceptible to ischemic injury during bypass and should be perfused at mean perfusion pressures higher than those for normotensive patients. Under conditions of hypothermia, a pH-stat strategy causes loss of cerebral blood flow autoregulation, and the cerebral blood flow becomes pressure passive. With both the pH-stat and alpha-stat strategies, cooling of the patient greatly increases the flow to metabolism ratio of the cerebral blood flow; however, this luxury perfusion brings to the brain not just an excess supply of oxygen but also an increased quantity of microemboli. Current investigative efforts are focused on the endothelial cell-leukocyte adhesion cascade, attempting to characterize beta2 and beta1 adhesion molecule expression in patients undergoing cardiac surgery. Hammersmith Hospital is about to complete a study of the effects of high-dose aprotinin on the inflammatory response pattern and on cerebral infarction. CONCLUSIONS: Further progress in the development of therapeutic and preventive strategies with respect to cerebral injury during cardiac bypass depends on an increase in the understanding of the mechanisms involved. Current strategies should include optimizing cerebral perfusion and minimizing macroembolic and microembolic damage. The possibility of modifying the systemic inflammatory response is the most interesting challenge of the next few years. PMID- 9869438 TI - Perceived parental/peer attitudes and alcohol-related behaviors: an analysis of the impact of the drinking age law. AB - This study examined the perceived change in parental and peer attitudes toward underage drinking associated with the raising of the legal drinking age and its effect on youthful alcohol use and drinking driving. The analysis was based on the New York State Youth Alcohol Survey, a series of four surveys conducted in 1982, before the enactment of the 19 drinking age law; 1983, after the enactment of the 19 drinking age law, 1985, before the 21 drinking law; and 1986, after the 21 drinking age law. Findings indicate that the effect of perceived parental attitudes is specific to underage alcohol use, but the effect of perceived peer attitudes is general to both underage and legal drinkers. Results from this study suggest that parental supervision is a key factor in enforcing the drinking age law and reducing youthful alcohol use. Parents' participation should be included in all enforcement and prevention/intervention efforts. PMID- 9869439 TI - Reasons for quitting and smoking temptation among adolescent smokers: gender differences. AB - Adolescent cigarette smokers from randomly selected classrooms from 24 California and Illinois high schools were assessed regarding their interest in cessation, reasons for quitting, and smoking temptation circumstances. These data were analyzed by gender. Males and females were not found to differ in quit stage or perceived likelihood of ever quitting smoking, although males reported being somewhat more likely to have ever tried to quit in the past. The associations of reasons for quitting were not found to vary by gender in most comparisons. On the other hand, the associations of smoking temptation circumstances with gender showed that a greater percentage of females than males reported more circumstances that would make them tempted to smoke. Smoking cigarettes to regulate one's affective states, and to avoid nicotine withdrawal, may be functions of smoking that impede efforts at quitting, particularly among adolescent females. PMID- 9869440 TI - Drug identity change processes, race, and gender. III. Macrolevel opportunity concepts. AB - Throughout the 20th century, scholars studying drug misuse have pinpointed various macrolevel (broad cultural and social structural) influences on the problem. The drug identity change model includes two such macrolevel factors. It maintains that social climates conducive to drug use (availability and popularity of drugs over the years) and social appraisal sources provide important opportunities for drug-related identity change. Social climates conducive to drug use help foster the formation of small groups centered on drug-related activity and meaning systems, i.e., social appraisal sources. Identification with these groups helps reduce one's ego identity discomfort or temporarily solve one's identity problems. It is through this process, the model contends, that young people change from nondrug users to drug misusers. Part 3 discusses these two concepts and how they work in the lives of the four race and gender groups studied here. PMID- 9869441 TI - Traditionalism in the prevention of substance misuse among Puerto Ricans. AB - Data are presented from a multicluster stratified sample of 1,084 Puerto Rican households in the New York City area. Researchers examined the impact of loss of traditional Hispanic family-oriented gender role values and degree of acculturation upon drug use. Sociodemographic variables are presented. Indices of "traditionalism" and "acculturation" were created, validated, and correlated with drug use. Traditionalism was found to covary negatively with drug use, but more so for women. The positive relationship between acculturation and drug use was found to be influenced by different levels of traditionalism. Implications for treatment and prevention are discussed. PMID- 9869442 TI - Impact of the 1970 legal BAC 0.05 mg% limit legislation on drunk-driver-involved traffic fatalities, accidents, and DWI in Japan. AB - Official statistics of motor vehicle fatality data have indicated that alcohol involvement in fatal crashes has declined substantially in Japan since 1970. The national campaign against drunken driving in Japan provides a natural experiment in which to test the predictions of deterrence theory. Utilizing official data over the 1960-1995 period, we report conclusive evidence that 1970s legislation is having a measurable and long-term effect on alcohol-related motor vehicle fatalities. Enactment of the lower legal blood alcohol limit with a combination of other severe sanctions has been more desirable in the prevention of alcohol related fatalities in the long term as shown in this study. Further extensive research is necessary to investigate performance at the lower legal limit in other societies in order to inspire a strong "international lobby" to support the reduced legal blood alcohol limit for drivers all over the world, as there is no doubt that a reduction in alcohol-impaired driving will result in a substantial savings of human lives and resource worldwide. PMID- 9869443 TI - Understanding the quality and outcome of treatment. PMID- 9869444 TI - The ordinary and the extraordinary: a note for an interpretative hypothesis. AB - The problem of the meaning to be given to the large changes that occur over the weekend in the way young people, and not so young people, live their lives is discussed. People who lead lives which can be defined as generically regular (as regards work, study, etc.) from Monday to Friday completely change their ways of being and living on Friday night. In order to emphasize, face, or sustain this change, drugs, especially designer drugs, are frequently used. The hypothesis is advanced that this may mean a rejection of the existential continuum that characterizes linear time and be a return to the phasic cyclical time attitude typical of ancient ages. Methods and attitudes for dealing with this problem are proposed. PMID- 9869445 TI - Changes in mortality, arrests, and hospitalizations in nonvoluntarily treated heroin addicts in relation to methadone treatment. AB - This study analyzes the mortality, hospitalizations, and arrests in a cohort of severe intravenous heroin users divided into three groups: those in methadone treatment, those discharged from treatment, and those who never received treatment. The study population consists of 101 heroin users, of whom 56 were HIV seropositive. Because of intensive drug misuse, they underwent coercive residential treatment in Stockholm during the 3-year period 1986-1988. The mortality was lower in the methadone group, and all seven deaths were related to HIV-infection. Outside the program, 24 of 29 persons died from external violence and poisoning. PMID- 9869446 TI - The impact of methadone on consumption of inpatient care and mortality, with special reference to HIV status. AB - This study investigates the impact of methadone treatment on inpatient care admissions and mortality among 331 methadone-maintained patients compared with 1,483 similar untreated opiate misusers. The methadone patients manifested a lower mortality during treatment than the comparison group and those patients who had left their treatment. The annual incidence rate decreased from 1.4 inpatient care admissions per year for those who had stayed 0-1 year, less than 1 for those who stayed more than 2 years, and 0.3 for those who had stayed longer (>4 years) in methadone treatment. A similar decrease occurred in both sexes. The incidence rate decreased more among the HIV-negative than the HIV-positive patients. The low incidence of inpatient care during treatment may be due to treatment but may also be partly due to selection factors. PMID- 9869447 TI - Identical variant TSG101 transcripts in soft tissue sarcomas and various non neoplastic tissues. AB - Inactivation of the TSG101 gene was recently shown to induce malignant transformation of NIH/3T3 fibroblasts. Abnormal TSG101 transcription profiles were observed in various human cancers, and large intragenic deletions of the TSG101 gene were reported for a series of human breast cancer specimens, pointing to a potential tumor-suppressive activity of TSG101. However, subsequent more detailed studies on a large panel of breast carcinoma samples did not confirm the tumor-associated genomic deletions. Here we analyzed the transcription patterns of the TSG101 gene in soft-tissue sarcomas and non-neoplastic human tissues. Forty-five of 71 soft tissue sarcoma samples (63%) displayed variant transcripts; however, identical aberrant transcripts were also detected in seven of 15 non neoplastic control tissues. Restriction fragment length polymorphism analysis of the TSG101 gene excluded major genomic rearrangements in the soft tissue sarcoma samples. Northern blot analysis revealed a very low abundance of variant transcripts as compared with the wild-type TSG101 transcript. These data point to aberrant splicing of the TSG101 mRNA in normal and transformed human mesenchymal tissues rather than tumor specific alterations of the TSG101 gene. In summary, this analyses does not support a pathogenic role for altered TSG101 expression in human soft tissue sarcomas. PMID- 9869448 TI - Arrest of replication by mammalian DNA polymerases alpha and beta caused by chromium-DNA lesions. AB - We have previously shown that trivalent chromium, and hexavalent chromium in the presence of one of its primary in vivo reductants, ascorbate, can bind to DNA and form interstrand crosslinks capable of obstructing replication. This effect was demonstrated in vitro by using Sequenase Version 2.0 T7 DNA polymerase; its parent enzyme, the unmodified T7 DNA polymerase; and Escherichia coli polymerase I large (Klenow) fragment; and it was demonstrated ex vivo by using Taq polymerase and DNA from chromium-treated human lung cells as template. This study was performed to determine whether DNA-bound chromium affects mammalian DNA polymerases in the same manner. Two mammalian enzymes, DNA polymerase alpha and DNA polymerase beta, were used. DNA polymerase alpha is a processive enzyme believed to be the primary lagging-stand synthetase, whereas DNA polymerase beta is a non-processive enzyme believed to function in DNA repair by filling single stranded gaps one base at a time. DNA polymerase arrest assays were performed with each of these enzymes to replicate DNA with toxicologically relevant levels of chromium adducts produced by either trivalent chromium or hexavalent chromium and ascorbate. Both enzymes responded to chromium-DNA damage by arresting replication, and the arrests increased in a dose-dependent manner. Furthermore, the guanine-specific pattern of arrests produced when an exonuclease-free preparation of DNA polymerase beta was used corresponded exactly to the arrest patterns produced in vitro by the exonuclease-free enzyme Sequenase and ex vivo by Taq polymerase. These results suggest that replication arrest may be a common response of polymerases to DNA-chromium lesions and provide a plausible mechanism for the inhibition of DNA synthesis and S-phase cell-cycle delay that occurs in mammalian cells treated with genotoxic chromium compounds. PMID- 9869449 TI - Complete cDNA sequence of the Ki-ras proto-oncogene in the liver of wild English sole (Pleuronectes vetulus) and mutation analysis of hepatic neoplasms and other toxicopathic liver lesions. AB - A complete copy of Ki-ras b cDNA from English sole (Pleuronectes vetulus), a benthic marine flatfish, was cloned and sequenced. The percent identity between the predicted amino acid sequence of English sole and human Ki-ras b was 97%, whereas the percent identity between the English sole gene and rainbow trout or Rivulus Ki-ras b was 98%. Areas of amino-acid sequence conservation included codons 12, 13, and 61, the positions in which mutations are observed in ras cellular oncogenes in other species. The 5' untranslated region (UTR), consisting of 217 nt, was not highly GC rich but contained four ATG start codons upstream of the major open reading frame. The 3' UTR, containing 26 nt, was AU rich. Analysis of Ki-ras mutations was performed on a variety of necrotic, preneoplastic, and neoplastic lesions in livers from 13 English sole collected from contaminated waterways in Puget Sound, WA. Despite reports of Ki-ras mutations in hepatic tumors from other fish, no mutations in codons 12, 13, or 61 were found in hepatic lesions from English sole by direct DNA sequencing of polymerase chain reaction-amplified genomic DNA. Although mutations could exist at levels below the detection limits of this analysis, the results suggest that Ki-ras has a role in liver carcinogenesis that varies according to the fish species or carcinogen. Furthermore, future studies of the etiology of chemically induced cancer in feral English sole should consider mutations in other cancer-related genes, such a5p53, Ha-ras, and N-ras. PMID- 9869450 TI - Regulation of keratinocyte growth factor expression in human endometrium: implications for hormonal carcinogenesis. AB - Estrogen is thought to be an important etiologic agent in endometrial and breast cancers. However, the mechanism or mechanisms by which estrogen acts as a hormonal carcinogen are not well understood. We hypothesize that in response to chronic exposure to estrogens, human endometrial stromal fibroblasts (ESF) produce factors that facilitate neoplastic transformation in epithelial cells. To test this hypothesis, we assessed the regulation of keratinocyte growth factor (KGF) mRNA and protein in ESF by interleukin-1 (IL-1) and diethylstilbestrol (DES). Short-term treatments with IL-1 but not with DES increased the abundance of KGF mRNA in ESF. However, chronic treatment with DES significantly increased KGF mRNA levels and protein production. KGF protein in medium conditioned by ESF chronically treated with 1 nM DES reached concentrations of approximately 100 ng/mL. At this concentration, KGF increased endometrial epithelial cell numbers fourfold and enhanced anchorage independence tenfold. These results suggest that KGF may play a role in hormonal carcinogenesis by mediating estrogen-induced changes in the interactions between stromal and epithelial cells. To address the potential role of nuclear transcription factor kappa B (NF-kappaB) in regulating KGF expression, we determined the effect of increased expression of its inhibitor, IkappaBalpha, on KGF mRNA and protein levels. Transfection with IkappaBalpha blocked induction of KGF expression by IL-1 but had no effect on the increase in KGF mRNA caused by chronic treatment with DES. These results suggest that IL-1 exerts its effects on KGF by an NF-kappaB-mediated pathway but that chronic treatment with DES stimulates KGF expression by some other mechanism. PMID- 9869451 TI - Alterations in cyclin-dependent kinase 2 function during differentiation of primary human keratinocytes. AB - Terminal differentiation of epithelial cells is intimately linked to cell-cycle withdrawal. The tight coupling of these two processes is critical to maintenance of epidermal tissue homeostasis and is frequently disrupted in squamous cell carcinoma. To identify possible molecular targets of epithelial carcinogenesis, we investigated the regulatory pathways that couple cellular differentiation and proliferation in primary cultures of human keratinocytes and found that the cyclin-dependent kinase inhibitors (CKIs) p21cip1/waf1 and p27kip1 were induced early during differentiation of human keratinocytes, whereas p15ink4B was induced later in differentiation. The induction of p21c1/waf1 was mediated by both transcriptional and non-transcriptional mechanisms, and the activities of cyclin A/cyclin-dependent kinase (cdk) 2 and cyclin E/cdk2 complexes were specifically inhibited during keratinocyte differentiation. In contrast, p21cip1/wafl did not associate with cdk4, and the activities of cdk4 complexes remained unchanged. Hence, our results support the model that multiple CKIs participate in linking cellular proliferation and differentiation in human keratinocytes by specific modulation of cdk2 activity. PMID- 9869453 TI - Allelotype analysis of uterine leiomyoma: localization of a potential tumor suppressor gene to a 4-cM region of chromosome 7q. AB - Uterine leiomyoma is a benign smooth muscle tumor of the myometrium and is the most commonly encountered neoplasm in women of reproductive age. As for most benign tumors, the pathogenesis of leiomyoma remains obscure, especially at the molecular genetic level. The purpose of this study was to perform a genome-wide allelotype analysis to identify potential sites of tumor suppressor gene inactivation. Fifty-two cases of uterine leiomyoma were subjected to allelotype analysis by using matched pairs of tumor and blood DNA. Loss of heterozygosity (LOH) was assessed at 61 microsatellite markers distributed throughout the genome and representing all 41 chromosome arms. In general, LOH was very rare except on chromosome 7q, where LOH was observed in 34% of all informative tumors. Fine deletion mapping with 25 microsatellite markers from the 7q22 region revealed a minimal deletion unit of approximately 4 cM, bounded by the markers D7S2453 proximally and D7S496 distally, that probably harbors a novel tumor suppressor gene involved in the etiology of this tumor. PMID- 9869452 TI - Unique expression patterns and alterations in the intestinal protein villin in primary and metastatic pulmonary adenocarcinomas. AB - The identification of markers that distinguish primary pulmonary adenocarcinomas from pulmonary adenocarcinomas secondary to the digestive tract would be clinically important. Villin, a specific marker in digestive-tract malignancies, was evaluated in 57 pulmonary adenocarcinomas, six samples of proximal bronchial tissue, and five metastatic pulmonary adenocarcinomas (three colon and two esophageal adenocarcinomas) by using immunohistochemical and molecular analyses. Villin was expressed in 31.6% (18 of 57) of the pulmonary adenocarcinomas and showed either a diffuse cytoplasmic pattern (10.5%) or a primary cytoplasmic pattern with minor brush-border staining (21.1%). However, none of those samples demonstrated the primary brush-border staining pattern that was characteristic of all five of the metastatic digestive-tract adenocarcinomas. There was a significant difference in the positive brush-border staining pattern between the primary and metastatic pulmonary adenocarcinomas (P < 0.002). Villin protein was expressed in bronchial epithelial cells, and villin mRNA was detected by reverse transcription-polymerase chain reaction. Northern analysis demonstrated 3.5- and 2.7-kb villin mRNAs in villin protein-positive tumors, but villin mRNA was not detected in non-tumorous lung tissue, indicating the transcriptional upregulation of villin in lung tumors. An additional smaller-sized mRNA (1.8 kb) was observed in six of 10 pulmonary adenocarcinomas and in the bronchoalveolar carcinoma cell line A549. Two small villin mRNAs were cloned from the cell line A549 and were found to represent an alternatively spliced (exon 8-exon 14) 1.85-kb mRNA and a 1.8-kb mRNA that was missing a portion of the 5' region (exon 1-exon 9) of the native villin mRNA. These studies demonstrated that the pattern of villin expression and the presence of altered villin mRNAs may be useful markers for pulmonary adenocarcinomas as well as provide support for the potential origin of villin-expressing tumors from bronchial epithelial cells. PMID- 9869454 TI - Loss of imprinting of the insulin-like growth factor II gene in mouse hepatocellular carcinoma cell lines. AB - We investigated expression of insulin-like growth factor II (Igf2) in primary cultured hepatocytes, liver epithelial (LE) cell lines derived from normal hepatocytes, and hepatocellular carcinoma (HCC) cell lines from crosses between C3H/HeJ (C3H) and Mus musculus molossinus mice (MSM). Igf2 mRNA was detected by reverse transcriptase-polymerase chain reaction in primary cultured hepatocytes from 5 d after the start of cultivation and in all 12 LE and 16 HCC cell lines. Analysis of the untranslated region of Igf2 exon 6, which contains polymorphic CA repeats, revealed that 13 of the 16 HCC cell lines had biallelic expression, whereas monoallelic expression was retained in the primary cultured hepatocytes and all 12 LE cell lines. The Igf2 transcripts contained exons 1-3 in all the HCC cell lines but only exons 2 and 3 in cultures of hepatocytes and LE cell lines, indicating difference in promoter use. However, the biallelic HCC cell lines did not have larger amounts of Igf2 mRNA and protein than did the monoallelic lines, suggesting that loss of imprinting may not be directly related to the level of Igf2 expression. PMID- 9869455 TI - Inhibition of intrinsic gap-junction intercellular communication and enhancement of tumorigenicity of the rat bladder carcinoma cell line BC31 by a dominant negative connexin 43 mutant. AB - The tumor-suppressive property of the connexin gap-junction proteins was postulated from the fact that their function of cell coupling is impaired in most cancer cells. However, in conflict with this notion, certain cancer cells are able to communicate through gap junctions despite their malignancy. To explain this phenomenon, we studied by using a dominant-negative strategy the effect on tumorigenicity of loss of intrinsic gap-junction intercellular communication (GJIC) in the rat bladder carcinoma cell line BC31, which shows both expression of connexin 43 (Cx43) and intercellular communication. In cells transfected with a mutant Cx43 with seven residues deleted from the internal loop at positions 130 136 (Cx43delta), transport of the resulting connexin protein to the plasma membrane occurred normally, but the GJIC of the cells was effectively abolished at the level of permeability of established gap junctions. Dominant-negative inhibition of GJIC by Cx43delta accelerated growth of BC31 cells in nude mice. In contrast, when GJIC in BC31 cells was artificially enforced by transfection of wild-type Cx43, the cells lost the capacity to grow in vivo. Decreased phosphorylation of Cx43delta suggested close interaction of the internal loop of connexin with its commonly phosphorylated domains in the C-terminal tail and involvement of this interaction in gap-junction permeability. Therefore, we conclude that the intrinsic GJIC observed in cancer cells should be considered a tumor-suppressor factor and that its level may influence malignant growth capacity. PMID- 9869456 TI - Long-term results after scleral buckling surgery in uncomplicated juvenile retinal detachment without proliferative vitreoretinopathy. AB - PURPOSE: To evaluate long-term anatomic and functional results of scleral buckling surgery in retinal detachment without proliferative vitreoretinopathy in juvenile patients. METHODS: A total of 33 eyes of 31 patients were operated on. Surgical techniques included segmental buckle (23 eyes), radial buckle (5 eyes), encircling band (4 eyes), and temporary balloon buckle (1 eye). The average age at time of surgery was 15.3 years and the average follow-up time was 8.9 years. Retinal detachment was associated with the following risk factors: myopia (14 eyes), blunt trauma (8 eyes), and aphakia (3 eyes). In 9 eyes, no risk factors were present. Fifteen eyes had a detached macula. In addition to anterior and posterior segment examination, binocular functions and ocular motility were evaluated. RESULTS: Retinal reattachment required one operation in 29 eyes, two operations in three eyes, and three operations in one eye. In one eye, pars plana vitrectomy had to be performed to reattach the retina. At the end of follow-up, retinal reattachment was achieved in all eyes. Visual acuity of 20/40 or better was achieved by 39.4% preoperatively and 60.6% postoperatively; in 52.4%, binocular functions were normal. CONCLUSION: Scleral buckling procedures allow favorable anatomic and functional results in juvenile retinal detachment without proliferative vitreoretinopathy. PMID- 9869457 TI - Retinal detachments due to traumatic tears in the pars plana ciliaris. AB - PURPOSE: To describe the clinical features of retinal detachments caused by traumatic tears in the pars plana ciliaris. METHODS: This study is a retrospective review of five cases of retinal detachments secondary to traumatic breaks limited to the nonpigmented epithelium of the pars plana ciliaris. Four patients had a history of recent closed-globe trauma due to a fist blow and the remaining patient had a history of remote closed-globe trauma. RESULTS: In all cases, the tearing of the pars plana developed in the superonasal quadrant. The low-lying configuration of the detachment and the absence of a retinal break, oral tear, or avulsed vitreous base challenged the ability of the initial examiners to establish the correct diagnosis. Each patient underwent successful surgical reattachment of the retina. CONCLUSIONS: Retinal detachments may result from traumatic tears in the nonpigmented epithelium of the pars plana ciliaris. These detachments often do not become clinically manifest for several months or longer following ocular contusion. They tend to be shallow with a smooth and flat surface, which makes diagnosis difficult during routine ophthalmoscopy. Scleral depression is almost always necessary to identify the pars plana tear(s). PMID- 9869459 TI - Scleral fixated intraocular lenses: an angiographic study. AB - PURPOSE: We reviewed the results, complications, and fluorescein angiographic (FA) findings in eyes that had undergone transscleral fixation of posterior chamber intraocular lenses (IOLs). METHODS: Posterior chamber IOL implantation with scleral fixation was performed on 18 patients. Three patients were aphakic, 14 had an intraoperative posterior capsule rupture during cataract surgery, and one was operated on for a retained lens nucleus and dislocated IOL in the vitreous. Follow-up examinations measured visual acuity, intraocular pressure, and IOL decentration and tilting. In 14 patients, iris and retinal FA were performed. RESULTS: No major complications were noted during the procedure. Mean best-corrected visual acuity was 20/70 preoperatively and 20/30 postoperatively after a mean follow-up of 9.8 months. Fourteen patients achieved a visual acuity of 20/40 or better. Macular epiretinal membranes were diagnosed after surgery in five eyes but only one eye showed significant distortion of the fovea (macular pucker). Iris FA revealed no major vascular abnormalities. Fluorescein angiography showed cystoid macular edema in six cases. Light-induced retinal lesions occurred in six eyes. CONCLUSIONS: Transscleral fixation of posterior chamber IOLs provided adequate visual acuity in most patients. Incomplete visual recovery after surgery may be related to the occurrence of macular edema and epiretinal membranes. Light-induced retinal injury was the major irreversible intraoperative complication. PMID- 9869458 TI - Cystoid macular edema following combined phacoemulsification and vitrectomy for macular hole. AB - PURPOSE: To evaluate the incidence of cystoid macular edema (CME) in patients undergoing either combined or consecutive pars plana vitrectomy (PPV) and phacoemulsification (PE) for the treatment of macular hole and cataract. METHODS: Retrospective review of a consecutive surgical series of 22 patients with idiopathic macular holes. Seven patients underwent combined PPV with air-fluid C3F8 gas exchange and PE with intraocular lens insertion; 15 patients underwent PE subsequent to PPV. RESULTS: Visual acuity improved by 2 or more lines in 3/7 patients in the combined group and in 15/15 in the consecutive group. Of seven patients in the combined group, two required a second PPV for treatment of the macular hole and one declined further treatment. Of importance, 3/7 in the combined group and 1/15 in the consecutive group developed clinical and angiographic CME, all treated successfully with sub-Tenon's triamcinolone injections. CONCLUSIONS: Both combined PPV/PE and consecutive PPV/PE offer excellent potential therapeutic benefits for the treatment of idiopathic macular holes. The incidence and pathogenesis of CME must be further investigated. PMID- 9869460 TI - Penetration of ofloxacin in human aqueous and vitreous humors following oral and topical administration. AB - PURPOSE: To determine aqueous and vitreous humor ofloxacin levels following oral and topical application of ofloxacin in patients with noninflamed cornea and intact crystalline lens, and to compare the drug levels provided by each route. MATERIALS AND METHODS: Twenty-six patients undergoing pars plana vitrectomy for various ocular pathologies were divided into two groups. Fourteen patients received two drops of 0.3% ophthalmic solution of ofloxacin every 30 minutes for 3 hours and every 60 minutes for the next 3 hours, and 12 patients received a single oral dose of 400 mg ofloxacin 8 hours before surgery. The aqueous and vitreous humor samples were simultaneously collected after oral or topical administration during pars plana vitrectomy to assess penetration of the drug. Samples were assayed for ofloxacin concentrations by a previously described method using high-performance liquid chromatography. RESULTS: The aqueous and vitreous humor levels of ofloxacin were 1.54 +/- 0.27 microg/mL (mean +/- standard error) and 1.77 +/- 0.24 microg/mL after oral and 1.44 +/- 0.24 microg/mL and 0.37 +/- 0.05 microg/mL after topical ofloxacin administration, respectively. Aqueous humor levels were not statistically different following oral or topical administration (P > 0.8). However, vitreous level of the drug after oral administration was significantly higher than that after topical administration (P < 0.001). CONCLUSION: Ocular bioavailability of ofloxacin in aqueous humor after oral and topical administration is similar when the drug is applied as described. Penetration of ofloxacin into vitreous humor is less than that into aqueous humor following topical application. The aqueous humor levels of ofloxacin via both routes and the vitreous level of the drug after oral route exceed the minimum inhibitory concentrations for certain bacterial species that frequently cause intraocular infection. PMID- 9869461 TI - Penetration of piperacillin into the vitreous after intravenous administration. AB - PURPOSE: Penetration of piperacillin into the vitreous cavity after intravenous administration was investigated in humans. METHODS: Forty-five eyes undergoing vitrectomy between November 1993 and December 1994 were included. Each patient received a single intravenous dose of piperacillin 4 g 2 hours before ocular incision. A 0.2-mL vitreous sample was aspired at the beginning of vitrectomy, a blood sample was obtained, and piperacillin level was assessed. RESULTS: There were no detectable drug concentrations in 25 eyes. Mean vitreous drug concentration in the remaining evaluable eyes (n = 14) was 2.33 microg/mL (+/ 0.76). We divided samples into two groups: 23 uninflamed (Group 1) and 16 inflamed (Group 2) eyes. Mean vitreous drug concentration was 0.42 microg/mL in Group 1 and 4.95 microg/mL in Group 2 (P < 0.2). Piperacillin concentration was higher than the minimum inhibitory concentration for gram-positive bacteria in 13% of uninflamed and 69% of inflamed eyes (P < 0.001). CONCLUSION: Studies show that intravenously administered piperacillin can penetrate the vitreous cavity in rabbits. Our study suggests that a single dose of piperacillin can produce intravitreal concentrations sufficient to kill gram-positive bacteria in inflamed eyes. The poor intraocular penetration of systemically administered piperacillin in uninflamed eyes raises questions about its usefulness as a prophylactic agent in ophthalmic surgery. PMID- 9869462 TI - ON-pathway dysfunction in a patient with acquired unilateral night blindness. AB - PURPOSE: To investigate possible functional correlates of an apparent ON-pathway defect observed in the cone electroretinogram of a patient with acquired unilateral night blindness. METHOD: Visual evoked potentials were recorded to the onset of a grid pattern consisting of either incremental or decremental squares. Saccadic eye movements were measured to luminance increments and decrements presented 5 degrees from fixation. The patient's results were compared with normative data. RESULTS: Visual evoked potential latencies were prolonged to incremental stimulation of the patient's affected left eye but were within normal limits for the other three conditions (increments and decrements, right eye; decrements, left eye). A similar pattern of asymmetry between latencies to incremental and decremental stimulation of the affected eye was observed for saccadic eye movements. CONCLUSIONS: The observed predominant delay in response to luminance increments supports the hypothesis of an ON-pathway dysfunction in this patient with acquired unilateral night blindness. PMID- 9869463 TI - Ocular and orbital blood flow in patients with essential hypertension treated with trandolapril. AB - PURPOSE: To study the effect of essential hypertension on flow velocity in the central retinal (CRA) and posterior ciliary arteries (PCA). Flow velocity was also evaluated in these arteries in patients with hypertension treated with trandolapril, an oral angiotensin-converting enzyme inhibitor. METHODS: Using the duplex scanner, flow velocity of the CRA and PCA was measured in 12 medication free patients with hypertension and 10 normal controls. The hypertensive patients were then treated with oral trandolapril, 1 mg/day for 1 week. After 1 week of treatment, flow velocity was again measured in the arteries of the patients with hypertension. RESULTS: There was a significant reduction in systolic and diastolic flow velocity of the vessels tested in the medication-free hypertensive patients when compared with those in the normal controls. In controls, the CRA had a peak systolic flow velocity (PSFV) of 34 cm/sec and an end diastolic flow velocity (EDFV) of 14 cm/sec; the PCA had a PSFV of 38 cm/sec and an EDFV of 16 cm/sec. In the hypertensive patients off medication, the CRA had a PSFV of 16 cm/sec and an EDFV of 6 cm/sec; the PCA had a PSFV of 17 cm/sec and an EDFV of 5 cm/sec. The diastolic component also was significantly decreased in the patients with hypertension. Flow velocity significantly increased in the hypertensive patients treated with trandolapril for 1 week, but did not reach the level of flow measured in normal controls. CONCLUSION: The decreased flow velocity in hypertensive patients may result from a peripheral vasospasm in the vessels of the eye and orbit. This decreased flow velocity may be important in eyes that already have ocular disease. Improvement in flow velocity was noted with oral trandolapril but it did not reach the levels seen in normal controls. PMID- 9869465 TI - Diagnostic and therapeutic challenges. PMID- 9869464 TI - C-C chemokines in the vitreous of patients with proliferative vitreoretinopathy and proliferative diabetic retinopathy. AB - PURPOSE: To investigate the presence of three C-C chemokines (monocyte chemotactic protein-1 [MCP-1], macrophage inflammatory protein-1alpha [MIP 1alpha], and MIP-1beta) in vitreous samples from eyes with proliferative vitreoretinopathy (PVR), proliferative diabetic retinopathy (PDR), or retinal detachment (RD). SUBJECTS AND METHODS: Vitreous samples were obtained and assayed by solid phase enzyme-linked immunosorbent assay for the chemokines MCP-1, MIP 1alpha, and MIP-1beta in a prospective study of 43 consecutive patients. Eighteen samples from cadaveric control eyes were also assayed. RESULTS: Monocyte chemotactic protein-1 was detected in all samples. The vitreous of eyes with vitreoretinal disorders showed significantly higher levels than the vitreous of cadaveric control eyes (P < or = 0.05). Median level (5th-95th percentile) in the PVR cases (n = 20) was 890.18 pg/mL (286.04-1806.20); in RD (n = 8), 296.69 pg/mL (171.44-1310.02); and in PDR (n = 15), 434.60 pg/mL (124.56-1092.94). In the cadaveric control eyes (n = 18), median level was 83.97 pg/mL (26.09-208.38). Macrophage inflammatory protein-1alpha and MIP-1beta were not detected in any samples. CONCLUSION: Monocyte chemotactic protein-1 might be involved in the recruitment of macrophages and monocytes into the vitreous of eyes with proliferative vitreoretinal disorders. PMID- 9869466 TI - Displacement of corneal cap during vitrectomy in a post-LASIK eye. PMID- 9869467 TI - Drainage of subretinal fluid with a radiosurgical instrument: a preliminary report. PMID- 9869468 TI - Combined hamartoma of the retina and retinal pigment epithelium: a bilateral presentation. PMID- 9869469 TI - Echographic characteristics of retinal arterial macroaneurysm. PMID- 9869470 TI - Successful surgical removal of encapsulated subretinal cysticercus. PMID- 9869471 TI - Combined tractional rhegmatogenous retinal detachment in familial exudative vitreoretinopathy associated with posterior retinal holes: surgical therapy. PMID- 9869472 TI - Cytomegalovirus retinitis and the lupus anticoagulant syndrome. PMID- 9869473 TI - A new way to visualize the macula during laser photocoagulation. PMID- 9869474 TI - External beam radiotherapy and choroidal neovascularization. PMID- 9869475 TI - Two news items. PMID- 9869476 TI - Noninvasive localization of human atherosclerotic lesions with indium 111-labeled monoclonal Z2D3 antibody specific for proliferating smooth muscle cells. AB - BACKGROUND: Targeting exclusive antigens in atherosclerotic plaques with antibodies may provide a noninvasive means to detect rapidly proliferative atherosclerotic lesions. 111In-labeled negative charge-modified Z2D3 F(ab')2 (Z2D3) specific for an antigen expressed exclusively by proliferating smooth muscle cells has been shown to accumulate in rabbit atherosclerotic plaques. METHODS: The safety, biodistribution, accumulation, and elimination of Z2D3 were assessed in 11 patients who were candidates for carotid endarterectomy. The presence of atheromas in these patients was confirmed by angiography and Doppler ultrasound. Z2D3 (250 microg) labeled with 5 mCi of 111In was administered by slow intravenous injection. Planar and single photon emission computed tomography (SPECT) images were obtained 4, 24, 48, and 72 hours later. Carotid endarterectomy was performed and the surgical specimens were imaged, weighed, gamma-counted, and analyzed by immunostaining. RESULTS: Uptake of Z2D3 at the site of the carotid plaques was observed in the planar and SPECT views at 4 hours in all subjects. In addition, antibody uptake was noted in the contralateral vessel in 5 subjects. SPECT images identified the atherosclerotic plaques with focal uptake. The antibody uptake corresponded with the angiographic location of the disease. Immunohistochemical studies of the endarterectomy specimens confirmed the localization of Z2D3 into the plaque areas containing smooth muscle cells. Adverse drug reactions were not observed. CONCLUSION: This study demonstrates the feasibility of targeting atherosclerotic lesions with negative charge-modified antibody. It also proposes the possibility of selective identification of various components of atherosclerotic plaque, which may contribute to determining strategies of intervention in future. PMID- 9869477 TI - Relation of myocardial perfusion at rest and during pharmacologic stress to the PET patterns of tissue viability in patients with severe left ventricular dysfunction. AB - BACKGROUND: Stress perfusion imaging can assess effectively the amount of jeopardized myocardium, but its use for identifying underperfused but viable myocardium has yielded variable results. We evaluated the relation between measurements of myocardial perfusion at rest and during pharmacologic stress and the patterns of tissue viability as determined by positron emission tomographic (PET) imaging. METHODS AND RESULTS: We studied 33 patients with coronary artery disease and left ventricular (LV) dysfunction (LV ejection fraction, 30%+/-8%). PET imaging was used to evaluate regional myocardial perfusion at rest and during pharmacologic stress with [13N]-ammonia as a flow tracer, and to delineate patterns of tissue viability (i.e., perfusion-metabolism mismatch or match) using [18F]-deoxyglucose (FDG). We analyzed 429 myocardial regions, of which 229 were dysfunctional at rest. Of these, 30 had normal perfusion and 199 were hypoperfused. A severe resting defect (deficit >40% below normal) predicted lack of significant tissue viability; 31 of 35 regions (89%) had a PET match pattern denoting transmural fibrosis. Although regions with mild or moderate resting defects (deficit <40% below normal) showed evidence of metabolic activity, perfusion measurements alone failed to identify regions with PET mismatch (reflecting hibernating myocardium). Reversible stress defects were observed with slightly higher frequency in regions with a PET mismatch (10 of 37) than in those with a PET match (36 of 162) pattern of viability. A reversible stress defect was a specific (78%) marker, but was a relatively insensitive marker (27%) of viable myocardium as defined by the PET mismatch pattern. CONCLUSIONS: In patients with LV dysfunction, the severity of regional contractile abnormalities correlates with the severity of flow deficit at rest. Severe reductions in resting blood flow in these dysfunctional regions identify predominantly nonviable myocardium that is unlikely to have improved function after revascularization. Although dysfunctional myocardium with mild to moderate flow reductions contains variable amounts of viable tissue (as assessed by FDG uptake), flow measurements alone do not distinguish between regions with PET mismatch (potentially reversible dysfunction) and PET match (irreversible dysfunction). The presence of an irreversible defect on stress imaging is a relatively specific (78%) marker of PET match, whereas a reversible stress defect is a rather insensitive (27%) marker of viability, as defined by the PET mismatch pattern. PMID- 9869478 TI - Quantitative gated blood pool SPECT for the assessment of coronary artery disease at rest. AB - BACKGROUND: Planar gated blood pool imaging (GBPI) has long proven to be useful for the noninvasive assessment of ventricular function. From a practical viewpoint, gated blood pool single photon emission computed tomography (GBPS) acquisition can be accomplished in the same time as a three-view planar series, with the benefit of a tomographic perspective that avoids chamber overlap. METHODS AND RESULTS: Quantitative gated blood pool SPECT was applied to 10 patients who underwent coronary arteriography, contrast ventriculography, and planar gated blood pool imaging. For each patient, the mid-short axis oblique slice was divided into 4 discrete segments using 4 different reference models and 2 forms of segmentation. A center of mass (counts) fixed in the end-diastolic frame and segmentation that bisected the ventricular septum proved to have the highest sensitivity and specificity for determining regional wall motion abnormalities at rest in myocardium supplied by severely diseased coronary arteries (>75 %). GBPS correctly identified 19 of 21 abnormal segments (90%), with good specificity (95%), whereas ventriculography identified 12 (57%) and planar GBPI identified 9 (43%) of the segments supplied by diseased coronaries. CONCLUSION: Quantitative GBPS appears to be a sensitive method for assessing coronary artery disease at rest in myocardium perfused by severely diseased coronary arteries. PMID- 9869479 TI - Validation of left ventricular volume measurements by gated SPECT 99mTc-labeled sestamibi imaging. AB - BACKGROUND: Previous studies have shown that gated single photon emission computed tomography (SPECT) technetium 99-labeled sestamibi imaging provides accurate and reproducible measurement of left ventricular (LV) ejection fraction (EF), wall motion, and thickening. This study examined the reliability of gated SPECT sestamibi imaging in measuring LV end-diastolic volume (EDV), end-systolic volume (ESV), and stroke volume (SV). METHODS AND RESULTS: Gated SPECT measurements were compared with an independent nongeometric method based on thermodilution SV and first-pass radionuclide angiographic EF (using a multicrystal gamma camera). Twenty-four patients aged 58+/-11 years underwent cardiac catheterization and coronary angiography for evaluation of chest pain syndromes. None had primary valvular disease, intracardiac shunts, or atrial fibrillation. RESULTS: The correlation between the two methods were as follows: EDV: r = 0.89, P<.001; ESV: r = .938, P<.001; SV: r = 0.577, P<.001. Bland-Altman plots showed mean differences (+/-standard deviation [SD]) for EDV of -14.3+/ 33.3 mL, for ESV of -0.4+/-23.7 mL, and for SV of -13.9+/-15.2 mL. The reproducibility of measuring EDV and ESV by gated SPECT was very high (r = 0.99 each). CONCLUSION: Gated 99mTc-labeled sestamibi SPECT provides reproducible LV volume measurements. With validation of volume measurement, gated SPECT provides comprehensive assessment of regional and global LV function. This information is important in many patient groups such as those with ischemic cardiomyopathy, concomitant coronary and valve disease, and those who have had myocardial infarction. It will also be useful to assess the incremental value of LV volumes in risk assessment. PMID- 9869480 TI - Cardiac death prediction and impaired cardiac sympathetic innervation assessed by MIBG in patients with failing and nonfailing hearts. AB - BACKGROUND: Although cardiac sympathetic nerve dysfunction is related to poor clinical outcome, a critical sympathetic dysfunction level for predicting cardiac death is still unclear. The current study was designed to investigate which indices derived from metaiodobenzylguanidine (MIBG) imaging have prognostic value compared with clinical and cardiac function variables, and to determine the threshold of cardiac MIBG activity for identifying patients likely to suffer cardiac death in both failing and nonfailing hearts. METHODS AND RESULTS: Myocardial I-123-MIBG activity was quantified as a heart-to-mediastinum ratio in 414 consecutive patients, 173 (42%) of whom had symptomatic heart failure. After cardiac function measurements, patients were followed up with an end-point of cardiac or noncardiac death. During a mean follow-up period of 22 months, 37 cardiac deaths occurred: 23 resulted from heart failure, 9 were sudden cardiac deaths, and 5 were fatal myocardial infarctions. Multivariate analysis using the Wald chi2 and the Cox proportional hazard model revealed that late heart-to mediastinum ratio, the use of nitrates, early heart-to-mediastinum ratio, and left ventricular ejection fraction were independent predictors of cardiac death; late heart-to-mediastinum ratio, New York Heart Association (NYHA) class, the presence of previous myocardial infarction, and age were independent predictors of heart failure and sudden cardiac death. Late heart-to-mediastinum ratio was the most powerful predictor of overall cardiac death among the variables. The Kaplan-Meier analysis showed that a late heart-to-mediastinum ratio of 1.74 or less, age greater than 60 years, the presence of myocardial infarction, and NYHA functional class 3 or 4 strongly indicated poor clinical outcomes. Furthermore, the more powerful incremental prognostic values were obtained by using MIBG imaging in combination with conventional clinical variables. CONCLUSIONS: Impaired cardiac sympathetic innervation assessed by MIBG activity has the greatest potential for predicting cardiac death and may be useful for identifying a threshold level for selecting patients at risk for death by heart failure, sudden cardiac death, and fatal myocardial infarction. PMID- 9869481 TI - 123I-MIBG myocardial scintigraphy as a noninvasive screen for the diagnosis of coronary artery spasm. AB - BACKGROUND: It has been suggested that the sympathetic nervous system might play an important role in the development of coronary artery spasm. However, no cardiac imaging modality has been able to demonstrate abnormal sympathetic innervation in patients with coronary artery spasm. The purpose of this study was to assess the presence and location of abnormal sympathetic innervation using iodine 123-metaiodobenzylguanidine (123I-MIBG) single photon emission computed tomography (SPECT) and to evaluate the clinical efficacy of 123I-MIBG SPECT as a noninvasive screening test in patients with coronary artery spasm. METHODS AND RESULTS: Coronary arteriography and a provocative test with intravenous administration of ergonovine maleate were performed in 26 patients (20 men, 6 women, mean age 48.2+/-12.0 years, range 20 to 67 years) who were suspected of having a coronary artery spasm. The subjects were divided into 2 groups: group 1 (n = 18) comprised subjects with a positive provocative test result, and group 2 (n = 8) comprised subjects with negative provocative test results. Ten healthy subjects served as controls. No abnormal MIBG uptake was observed in the control subjects. Abnormal sympathetic nervous innervation using 123I-MIBG SPECT was observed either as a reduced uptake or a defective pattern in the perfused areas in 13 of the 18 regions supplied by vessels of ergonovine-induced vasospasm. Normal sympathetic innervation, as evidenced by normal 123I-MIBG uptake, was noted in all of the 60 segments of normal vessel territories. Reduced uptake of 123I-MIBG was not detected in the perfused areas of 5 vasospasm-induced vessels (perfusion territory of left anterior descending coronary artery [LAD] and the right coronary artery [RCA] in 2 and 3 patients, respectively). The sensitivity and specificity of 123I-MIBG for detection of coronary artery spasm were 72.2% (95% confidence interval [CI] 55% to 89%) and 100%, respectively. The positive predictive and negative predictive values were 100% and 92.3% (95% CI 91% to 93%), respectively. CONCLUSION: 123I-MIBG SPECT is a feasible method to evaluate noninvasively and localize the territories of coronary arteries with spasm. Invasive diagnostic coronary arteriography with ergonovine provocation test may be unnecessary for diagnosis of coronary artery spasm in patients with typical resting pain, negative exercise test or normal thallium perfusion scan results, but showing abnormalities in 123I-MIBG SPECT. PMID- 9869482 TI - Evolving therapeutic concepts and imaging in ischemic cardiomyopathy. PMID- 9869484 TI - Acute myocardial perfusion imaging for the technologist. PMID- 9869483 TI - Regulation of gene expression: how can molecules make the right ventricle distinct from the left? PMID- 9869485 TI - Z2D3 imaging: fringe or frontier? PMID- 9869486 TI - Myocardial viability: seeking relevance and redefinition. PMID- 9869487 TI - The VANQWISH Trial: support for the noninvasive strategy for risk stratification after acute myocardial infarction. PMID- 9869490 TI - Na-H exchange inhibition with cariporide limits functional impairment caused by repetitive ischemia. AB - Intracellular calcium ([Ca]i) overload on reperfusion may be one of the mechanisms responsible for ischemia-induced regional myocardial dysfunction. Because inhibiting the Na-H exchanger (NHE) limits intracellular sodium ([Na]i) and subsequent [Ca]i accumulation, we hypothesized that NHE inhibition would attenuate regional dysfunction in response to 25 cycles of ischemia (I, 2-min) and reperfusion (R, 8-min) of the left circumflex coronary artery (LCx) in conscious swine. Six animals were instrumented to measure arterial pressure, regional myocardial blood flow (colored microspheres), systolic wall thickening (WTh) in the normally perfused (left anterior descending, LAD) and LCx regions (sonomicrometry), LCx blood flow velocity (Doppler), and to reversibly occlude the LCx (hydraulic occluder). Each animal completed three protocols separated by 7 days: ISC, 25 I/R cycles; CAR, 25 I/R cycles + NHE inhibition (cariporide); and VEH, vehicle administration for 4.2 h. Regional myocardial blood flow was measured during LCx occlusion in the first protocol and 10 min after I/R 25 in all protocols. Systemic hemodynamics were similar among and within each protocol. Blood flow measured during LCx occlusion confirmed that perfusion was reduced (p < 0.05) to this compared with the LAD region. During ISC, LCx WTh was reduced (p < 0.05) after five IR cycles, and a stable reduction (approximately 55% of baseline; p < 0.05) was present after 20 I/R cycles. During CAR, LCx systolic WTh was reduced (p < 0.05) only after 15 and 25 I/R cycles (approximately 80 and 72%, respectively). The decrease in LCx WTh was greater in ISC than in CAR (p < 0.05). LCx WTh was not altered during VEH, while LAD WTh was similar within and among all protocols. Regional blood flow measured after 25 I/R cycles was not different among protocols. Our results indicate that NHE inhibition delays the onset and limits the degree of regional dysfunction in response to repeated bouts of ischemia and reperfusion. PMID- 9869488 TI - Abnormal thallium-201 chloride myocardial SPECT findings with noncritical coronary angiography in a patient after coronary artery bypass graft. PMID- 9869491 TI - Sodium channel-blocking properties of spiradoline, a kappa receptor agonist, are responsible for its antiarrhythmic action in the rat. AB - Spiradoline (U-62,066E), a selective kappa (kappa) receptor agonist, was examined for actions on the cardiovascular system and on myocardial ionic currents in rats. We initially characterized cardiac, hemodynamic, and antiarrhythmic actions of spiradoline in isolated perfused rat hearts and pentobarbital-anesthetized rats. Electrophysiologic studies in isolated myocytes were used to elucidate the mechanism for changes observed in vivo in the ECG, as well as for antiarrhythmic actions against electrical and ischemia-induced arrhythmias. In isolated rat hearts, spiradoline reduced heart rate and cardiac contractility and increased the PR interval and QRS width of the ECG in a concentration-dependent manner. In anesthetized rats, spiradoline dose-dependently reduced blood pressure and heart rate and prolonged the PR interval and QRS width. At slightly higher doses, it increased the QaT interval of the ECG. RSh, an index of sodium channel blockade in the rat, also was dose-dependently increased. Electrical stimulation of the left ventricle suggested that spiradoline may exert its antiarrhythmic action by blockade of myocardial sodium currents. The electrophysiologic actions of spiradoline on sodium currents, the transient outward (i(to)) and sustained plateau potassium (ik(sus)) currents were studied in isolated cardiac rat myocytes by whole-cell patch-clamp techniques. Spiradoline (15-500 microM) reduced peak sodium current in a rapid, reversible, and concentration-dependent manner; it also increased the rate of decay of I(to) and reduced the amplitude of Ik(sus). At a concentration of 150 microM, spiradoline produced a 24 +/- 2 mV hyperpolarizing shift in sodium current inactivation kinetics but did not alter activation processes. Spiradoline showed both tonic and frequency-dependent components of sodium current block. Thus spiradoline produced its antiarrhythmic actions via sodium channel blockade in myocardial tissue, although higher doses also block potassium currents. This combined ion channel-blocking property may be of added clinical benefit in the setting of myocardial ischemia. PMID- 9869492 TI - Electrophysiologic effects of civamide (zucapsaicin) on canine cardiac tissue in vivo and in vitro. AB - The cardiac electrophysiologic effects of civamide (zucapsaicin), the cis-isomer of the alkyl vanillylamide, capsaicin, were evaluated in intact dogs and isolated Purkinje fibers. In anesthetized dogs, the mechanism of ventricular tachycardia inducible from 1 to 3 h after coronary artery occlusion was determined by activation mapping. Of 16 dogs studied, nine had ventricular tachycardia of focal endocardial origin; four, a reentrant mechanism; and three had no inducible arrhythmia. Civamide (50 microg/kg) was administered to 10 of 13 dogs that were inducible, but three dogs were used as time controls. Transmural activation times were unaltered by civamide, but mean arterial pressure decreased from 76 +/- 10 to 66 +/- 10 mm Hg (p < 0.05), and muscle refractory periods shortened from 138 +/- 3 to 132 +/- 4 ms (p < 0.05). Civamide altered inducibility in five of six dogs with ventricular tachycardia of focal endocardial origin, but those with epicardial reentrant mechanisms were not affected in three of four dogs. With microelectrode techniques in vitro, civamide (10(-5) M) shortened the action potential duration at 50% repolarization (APD50) from 193 +/- 13 to 177 +/- 12 ms (p < 0.01) and APD90 from 260 +/- 15 to 248 +/- 13 ms (p < 0.01) in isolated Purkinje fibers (n = 10). Nifedipine prevented the effects of civamide in vitro. These results show that civamide may alter inducibility of ventricular tachycardia with focal endocardial origin and shorten APD of Purkinje fibers in vitro. The effects of civamide in vitro are prevented by preexposure of the Purkinje fibers to nifedipine, suggesting that the electrophysiologic effects of civamide may be mediated through blockade of calcium channels. PMID- 9869493 TI - Reduced periinfarction mortality as a result of long-term therapy with captopril but not hydralazine or propranolol in spontaneously hypertensive rats. AB - Hypertension and left ventricular hypertrophy (LVH) are known to increase susceptibility to ventricular arrhythmias during and before myocardial ischemia and to increase the risk of periinfarction mortality. Although regression of LVH has been advocated as a therapeutic goal, little evidence exists to suggest that it can reduce periinfarction mortality, and if it does, by which mechanisms it may do this. In this study, we evaluated the effects of control of systemic arterial blood pressure, of regression of myocardial hypertrophy, and of cardiac fibrosis on the susceptibility to ventricular arrhythmias and periinfarction mortality in the spontaneously hypertensive rat (SHR) model of hypertension and LVH. After 12 weeks of treatment, captopril and hydralazine reduced systolic blood pressure to 93 +/- 14 and 126 +/- 13 mm Hg, respectively, as compared with 193 +/- 12 mm Hg, p < 0.05, in the untreated control SHR group. The decrease with propranolol (to 185 +/- 12 mm Hg) was of borderline significance. There was a significant decrease in inducibility of ventricular arrhythmias by programmed electrical stimulation with captopril (5%; p < 0.05). One hour after infarction, there was a trend toward reduced mortality in the rats treated with hydralazine, 9.5% (p = 0.20 vs. control; p = 0.10 vs. propranolol), and captopril, 5% (p = 0.08 vs. control; p = 0.010 vs. propranolol). However, only captopril reduced 3-h postinfarction mortality (40%; p = 0.022) compared with 72% in the control group. The results showed a significant decrease of the left ventricular weight/body weight ratio in the rats treated with hydralazine (2.6 +/- 0.2 mg/g; p < 0.05) and captopril (2.2 +/- 0.2 mg/g; p < 0.05) compared with the control group (2.8 +/- 0.2 mg/g). An assessment of cardiac fibrosis indicated that captopril decreased the volume percentage of collagen the most (2.01 +/- 0.53; p < 0.05), followed by propranolol (2.29 +/- 0.64; p < 0.05) and hydralazine (2.92 +/- 0.58; p < 0.05) versus controls (3.23 +/- 0.61). This study suggests that regression of myocardial hypertrophy or long-term normalization of arterial systolic blood pressure or both are the major determinants of very early mortality (within 1 h after infarction) and that later mortality (3 h after infarction) may be the result of a more complex interplay of regression of myocardial hypertrophy and fibrosis and of control of blood pressure. PMID- 9869494 TI - Influence of age and dietary restriction on norepinephrine uptake into cardiac synaptosomes. AB - The purpose of this study was to determine whether aging alters neuronal uptake of norepinephrine (NE) in the rat heart and if dietary restriction influenced the effect of age on this system. Cardiac synaptosomes were prepared from 6-, 12- and 24-month-old male F344 rats fed ad libitum (AL) or a diet restricted (DR) to 60% of AL intake. Cardiac synaptosomes were incubated with 50, 100, 200, or 400 nM [3H]NE for 10 min at 37 degrees C with and without desmethylimipramine (DMI), a selective neuronal-uptake blocker. DMI-sensitive [3H]NE uptake was calculated as the difference between samples with and without DMI. NE uptake was adjusted for the number of cardiac synaptosomes in each sample by dividing by the endogenous NE content in each sample. The Vmax for uptake ([3H]NE/min/ng NE) declined significantly between 6 and 12 months in AL rats and between 12 and 24 months in DR rats. Km was not significantly different between age or diet groups. The change in Vmax with age suggests that the number of NE transporters per synaptosome may decline with age and that DR delays this effect of age. There were no differences in the sensitivity to DMI between age or diet groups. PMID- 9869495 TI - Chronic dual inhibition of angiotensin-converting enzyme and neutral endopeptidase during the development of left ventricular dysfunction in dogs. AB - Angiotensin-converting enzyme (ACE) inhibition as well as neutral endopeptidase (NEP) inhibition was demonstrated to influence hemodynamics in various cardiac disease states. However, specific effects of chronic combined ACE and NEP inhibition on left ventricular (LV) and myocyte geometry and function remain unclear. In this study, a dual-acting metalloprotease inhibitor (DMPI), which possesses both ACE and NEP inhibitory activity, was used in a rapid-pacing model of LV dysfunction. LV and myocyte geometry and function were examined in control dogs (n = 6), in dogs with pacing-induced LV dysfunction (216 +/- 2 beats/min, 28 days, n = 7), and in dogs with DMPI treatment during rapid pacing (10 mg/kg p.o., b.i.d., n = 6). With chronic rapid pacing, LV end-diastolic volume increased (84 +/- 4 vs. 49 +/- 3 ml), and LV ejection fraction decreased (38 +/- 3% vs. 68 +/- 3%) compared with control (p < 0.05). DMPI concomitantly administered during long term rapid pacing did not change LV ejection fraction (35 +/- 3%), but LV end diastolic volume was reduced (70 +/- 5 vs. 84 +/- 4 ml; p < 0.05) when compared with rapid pacing only. With long-term rapid pacing, myocyte cross-sectional area was decreased (278 +/- 5 vs. 325 +/- 5 microm2), and resting length increased (178 +/- 2 vs. 152 +/- 1 microm) when compared with control (p < 0.05). With DMPI concomitantly administered during rapid pacing, myocyte cross-sectional area (251 +/- 5 microm2) and resting length (159 +/- 4 microm) were reduced when compared with rapid pacing only (p < 0.05). Myocyte velocity of shortening decreased from control values with long-term rapid pacing (39.3 +/- 3.9 vs. 73.2 +/- 5.9 microm/s; p < 0.05) but improved with DMPI treatment during rapid pacing when compared with rapid pacing only (58.9 +/- 6.7 microm/s; p < 0.05). Myocyte velocity of shortening with beta-adrenergic-receptor stimulation (25 nM isoproterenol) was reduced from controls with rapid pacing (125 +/- 12 vs. 214 +/ 30 microm/s; p < 0.05) but was improved with DMPI treatment during rapid pacing when compared with rapid pacing only (178 +/- 12 microm/s; p < 0.05). In a model of rapid pacing-induced LV failure, concomitant DMPI treatment significantly reduced the degree of LV dilation with no apparent effect on LV pump function. At the level of the LV myocyte, long-term DMPI treatment with rapid pacing improved myocyte performance and beta-adrenergic response. Thus the improvement in isolated myocyte contractile function was not translated into improved global LV pump performance. The mechanisms by which improved myocyte contractility was not translated into a beneficial effect on LV-pump function with DMPI treatment during rapid pacing remain speculative, but likely include significant changes in LV remodeling and loading conditions. PMID- 9869496 TI - Long-term alpha1-adrenergic blockade attenuates diet-induced dyslipidemia and hyperinsulinemia in the rat. AB - This study evaluated the ability of alpha1-adrenergic blockade to interfere with the development of diet-induced hyperlipidemia and deterioration of insulin action. Diets having extremely divergent effects on glucose and lipid metabolism were contrasted. Rats were fed for 4 weeks either a nonpurified diet (chow) or a hyperlipidemic (HL) purified diet containing 40% energy as sucrose, 40% as fat, and 20% as casein. Half of each dietary cohort was given the alpha1-adrenergic antagonist prazosin (3 mg/kg/day in the food). Blood was collected in the fasted state (10 h after food removal) and 2 h after the intake of a meal. In the fasted state, plasma triacylglycerols (TGs) were higher in rats fed the HL diet than in those given chow and were not affected by long-term treatment with prazosin. Postprandially, plasma TG increased twofold in the chow-fed group, with or without long-term prazosin. In contrast, prazosin reduced by more than half the eightfold increase in TG that followed intake of the high-fat meal (Diet x Blocker interaction; p < 0.002) in the HL cohort. The HL-fed animals also displayed fasting hypercholesterolemia (+30%; p < 0.0001), which was prevented by long-term treatment with prazosin. Likewise, the 50% increase in plasma cholesterol that followed meal ingestion only in the HL cohort was blunted by the alpha1-blocker (Diet x Blocker interaction; p < 0.001). Long-term prazosin also abolished fasting hyperinsulinemia in the HL cohort, whereas it did not alter fasting insulin in chow-fed animals (Diet x Blocker interaction; p < 0.005). Measurement of postprandial lipoprotein lipase activity in several tissues did not suggest the involvement of changes in the absolute availability of the enzyme as a determinant of the hypotriacylglycerolemic action of the alpha1-blocker. Thus long-term alpha1-adrenergic blockade, with minimal effects in rats fed a hypolipidemic diet, strongly attenuates several of the fasting and postprandial alterations in plasma variables of lipid and glucose metabolism induced by an extremely lipogenic diet. PMID- 9869497 TI - Usefulness of transdermal clonidine in hypertensive patients undergoing minor surgical operations. AB - Transdermal clonidine (TTSC) treatment was evaluated in 29 patients with mild to moderate hypertension scheduled for minor surgery. Two weeks before the scheduled operation, patients underwent 24-h ambulatory blood-pressure monitoring (ABPM) to evaluate the efficacy of previous oral antihypertensive treatment, which was then substituted with TTSC, 0.1 mg/day. After 1 week, the efficacy of TTSC was clinically assessed, and the dose increased to 0.2 mg/day if needed. ABPM was repeated 2 days before the scheduled operation and 2 days after surgery. The 24-h blood pressure (BP) and heart rate (HR) profiles were smoothed by Fourier analysis. Three patients withdrew for adverse events and one for inefficacy after dose adjustment, TTSC being effective in the remaining 25 patients. Two patients who completed treatment lacked postsurgical ABPM recording. In the 23 patients with all ABPM recordings, average 24-h BP and HR obtained preoperatively during TTSC treatment were slightly reduced compared with values recorded during previous oral therapy. BP changes after surgery were negligible, whereas HR showed a moderate increase. Minor adverse events occurred in four (14%) of 29 patients. Our results demonstrate that TTSC provides adequate BP control in patients with mild to moderate hypertension undergoing minor surgery. PMID- 9869498 TI - In vivo left ventricular function and collagen expression in aldosterone/salt induced hypertension. AB - Cardiac fibrosis is linked to aldosterone-induced hypertension, but the effects on in vivo left ventricular (LV) function are not established. We studied the relations between in vivo LV function and aldosterone/salt cardiac fibrosis. Adult guinea pigs (GPs) were treated for 3 months with an aldosterone infusion and high-salt diet. This treatment induced arterial hypertension (+35%) and moderate LV hypertrophy (LVH; +60%) without right ventricular (RV) hypertrophy. Echo-Doppler LV assessment demonstrated unaltered cardiac output, stroke volume, or LV relaxation. Type I collagen messenger RNA (mRNA) was significantly increased in both ventricles (LV, +48%; RV, +77%) and accompanied by a significant increase in total collagen deposition (LV, from 0.52% in controls to 4.4% in treated GPs; RV, from 0.82 to 5.5% in treated GPs). Plasma norepinephrine levels increased 2.6-fold (p < 0.01) and correlated with the increase in collagen deposition in both ventricles. Collagen content was not correlated with hypertension or LVH. We conclude that aldosterone administration induces cardiac collagen accumulation and a sympathetic stimulation, which might preserve systolic and diastolic function. PMID- 9869499 TI - PD 142893, SB 209670, and BQ 788 selectively antagonize vascular endothelial versus vascular smooth muscle ET(B)-receptor activity in the rat. AB - The purpose of this study was to determine whether vascular endothelial and vascular smooth-muscle endothelin ET(B) receptors could be quantitatively differentiated by PD 142893 (PD), SB 209670 (SB), and BQ 788 (BQ) in the same species by using closely matched experimental conditions. The isolated perfused rat kidney (vascular smooth muscle) and isolated perfused rat mesentery (vascular endothelium) were challenged with increasing bolus doses of sarafotoxin S6c in the absence and presence of antagonist. PD, SB, and BQ produced parallel concentration-dependent rightward shifts in the S6c dose-response curve in the kidney. PD and SB also produced parallel concentration-dependent rightward shifts in the S6c dose-response curve in the mesentery. In contrast, BQ produced an insurmountable antagonism. Schild-derived pA2 values for PD and SB were significantly greater for inhibiting endothelial versus smooth-muscle ET(B) receptors. Furthermore, PD and SB differed in their relative potency between the two assays. Because BQ produced an insurmountable antagonism in the mesentery, it was not possible quantitatively to compare the antagonist activity in the two assays. These results indicate that PD, SB, and BQ selectively antagonize endothelial ET(B)-receptor activity over smooth-muscle ET(B)-receptor activity. PMID- 9869500 TI - Endothelium-dependent hyperpolarization in isolated arteries taken from animals treated with NO-synthase inhibitors. AB - To study the effects of chronic in vivo inhibition of NO synthase on endothelium dependent hyperpolarization, cell-membrane potential (in individual vascular smooth-muscle cells) and changes in tension (in isolated rings) were recorded from isolated canine coronary arteries and guinea-pig carotid arteries and aortas. In coronary arteries taken from control dogs and contracted with U46619, acetylcholine- and bradykinin-induced endothelium-dependent relaxations, which were unaffected by short-term in vitro exposure to indomethacin but were inhibited partially by L-nitro-arginine (LNA). In coronary arteries taken from dogs treated over the long term in vivo with LNA (30 mg/kg on the first day and 20 mg/kg the 7 following days, i.v.), the response to acetylcholine and bradykinin was inhibited when compared with arteries from control dogs. Short term in vitro exposure to LNA or indomethacin or both did not influence the effects of either agonist. In these arteries, the hyperpolarizing response to acetylcholine, observed in the presence of LNA and indomethacin, was enhanced, whereas that to bradykinin was partially inhibited. In the guinea pig isolated aorta, the relaxation to bradykinin was abolished by long-term in vivo treatment with L-nitro-arginine-methyl-ester (L-NAME; 1.5 mg/ml, in the drinking water for > or =4 days). In the isolated guinea pig carotid artery studied in the presence of LNA and indomethacin, acetylcholine induced a hyperpolarization that was not significantly affected by long-term in vivo treatment with L-NAME. These findings indicate that endothelium-dependent hyperpolarizations are maintained during long term inhibition of NO synthase and probably act as a back-up mechanism to elicit endothelium-dependent relaxations. PMID- 9869501 TI - Endogenous angiotensin II and bradykinin delay and attenuate the hypotension after N-type calcium channel blockade in conscious rabbits. AB - The effects of N-type calcium channel inhibition with omega-conotoxin GVIA (omega CTX) on cardiovascular parameters and vagally mediated autonomic reflexes and the role of the renin-angiotensin system were assessed in conscious rabbits. Omega CTX (10 microg/kg, i.v.) resulted in hypotension, tachycardia, and attenuation of the sympathetic and vagal components of the baroreceptor-heart rate reflex (baroreflex). In the control group (no pretreatment), the peak decrease in mean arterial pressure (MAP) of 13 +/- 3 mm Hg from 72 +/- 2 mm Hg occurred after 33 +/- 3 min, with a corresponding tachycardia of 80 +/- 20 beats/min (n = 6). The tachycardia was due to vagal withdrawal, as a similar increase in heart rate (84 +/- 8 beats/min) after omega-CTX was observed after pretreatment with the beta adrenoceptor antagonist, propranolol (n = 6). Angiotensin-converting enzyme (ACE) inhibition with enalaprilat revealed a larger, more rapid decrease in MAP in response to omega-CTX (-19 +/- 4 mm Hg from 65 +/- 1 mm Hg after 18 +/- 2 min; n = 6) compared with the control group. Similar larger decreases in MAP were also observed in the presence of the AT1-receptor antagonist, losartan, or the bradykinin B2 receptor antagonist, HOE-140 (n = 5-6). Pretreatment with enalaprilat, losartan, or HOE-140 caused a 50% decrease in the reflex tachycardia after omega-CTX compared with that observed in the control group, and omega-CTX caused a greater attenuation of the vagal component of the baroreflex and a decrease in the bradycardia evoked by the Bezold-Jarisch-like reflex. Also, there was a significant decrease in the bradycardia induced by the nasopharyngeal reflex after omega-CTX in the presence of ACE inhibition and HOE-140. Thus in the conscious rabbit, angiotensin II and bradykinin have a role in attenuating and slowing the hypotensive effect of N-type calcium channel inhibition. Vagolytic effects of omega-CTX on the baroreflex are augmented, and on other vagal reflexes are unmasked, via inhibition of the renin-angiotensin system. The complexity and mechanism of the interaction between N-type calcium channels and the renin angiotensin system remain to be elucidated. PMID- 9869502 TI - Pharmacologic preconditioning induced by beta-adrenergic stimulation is mediated by activation of protein kinase C. AB - Ischemic preconditioning (I-PC) occurs via activation of protein kinase C (PKC). This study was undertaken to determine whether pharmacologic preconditioning by beta-adrenergic stimulation (beta-PC) is mediated by PKC activation. Isolated rat hearts were subjected to 40-min ischemia and 30-min reperfusion. Beta-PC was induced by 0.25 microM isoproterenol pretreatment for 2 min followed by 10-min normoxic perfusion. Beta-PC enhanced the recovery of rate-pressure product of the ischemic/reperfused heart (79.1 +/- 8.4% vs. 12.4 +/- 1.6% of initial for Non-PC group, n = 6) and attenuated the release of creatine kinase during 30-min reperfusion (30.2 +/- 2.2 vs. 59.8 +/- 6.1 nmol/min/g wet wt for Non-PC group, n = 6), similar to an I-PC stimulus of 5-min ischemia and 5-min reperfusion. Treatment with 50 microM polymyxin B, a PKC inhibitor, abolished the cardioprotection of both beta-PC and I-PC. Furthermore, similar changes in subcellular distribution of PKC were induced by both beta-PC and I-PC. The changes in subcellular distribution of PKC-delta suggested its translocation from cytosol to membrane fraction, a marker of PKC activation. These results suggest that the cardioprotection induced by beta-PC, like I-PC, is mediated by PKC activation. PMID- 9869504 TI - Bunazosin hydrochloride inhibits exaggerated growth of vascular smooth muscle cells from spontaneously hypertensive rats by suppressing the response to growth factors. AB - Selective alpha1-adrenoreceptor blockers were recently reported to have an in vivo antiproliferative effect on hypertensive cardiovascular organs. Cultured vascular smooth-muscle cells (VSMCs) from spontaneously hypertensive rats (SHRs) show exaggerated growth compared with cells from Wistar-Kyoto (WKY) rats. We investigated the effects of an alpha1-adrenoreceptor blocker, bunazosin hydrochloride (HCl), on the growth of VSMCs from SHRs. In the absence of serum, bunazosin HCl significantly inhibited basal DNA synthesis by VSMCs from SHRs, but not by cells from WKY rats. In the presence of serum, bunazosin HCl significantly inhibited DNA synthesis by VSMCs from both rat strains. Angiotensin (Ang) II, platelet-derived growth factor (PDGF)-AA, and epidermal growth factor (EGF) dose dependently increased DNA synthesis by VSMCs from SHRs, but not by VSMCs from WKY rats. Bunazosin HCl significantly suppressed the response of DNA synthesis to PDGF-AA and EGF, but not to Ang II, in VSMCs from SHRs. Expression of basic fibroblast growth factor (bFGF), transforming growth factor-beta1 (TGFbeta1), and PDGF messenger RNA (mRNA) was markedly greater in VSMCs from SHRs than in cells from WKY rats. Bunazosin HCl significantly inhibited the expression of bFGF and TGFbeta1 mRNA in VSMCs from SHRs, but not in cells from WKY rats. These findings suggest that the inhibition of growth factor hyperresponsiveness and inhibition of the expression of growth factors in VSMCs from SHRs are associated with the antiproliferative effect of bunazosin. PMID- 9869503 TI - Cardiac and hemodynamic effects of the sinus node inhibitor tedisamil dihydrochloride in patients with congestive heart failure due to dilated cardiomyopathy. AB - Clinical and experimental investigations have demonstrated an inverse relation between heart rate and myocardial performance in patients with congestive heart failure. Accordingly, this study was designed to investigate the hemodynamic effect of the novel bradycardic compound tedisamil in patients with heart failure. We hypothesized that tedisamil would reduce heart rate and thereby improve hemodynamic parameters of failing hearts with an inverse force-frequency relation. Tedisamil was administered intravenously in nine patients with dilated cardiomyopathy (NYHA II-III). Hemodynamic measurements by right heart catheterization were carried out at time points -30, 10, 20 min, 1, 2, 4, and 6 h. Tedisamil decreased heart rate significantly from 84 +/- 6 beats/min to 73 +/- 4 beats/min (at 10 min; p < 0.05). Stroke volume index remained unchanged, and cardiac index tended to decrease transiently. Mean blood pressure increased from 98 +/- 5 to 104 +/- 6 mm Hg (p < 0.05) because of an increase in systemic vascular resistance from 1,619 +/- 145 to 2,079 +/- 198 dyn x s x cm(-5) (at 20 min; p < 0.05). Diastolic pulmonary pressure and pulmonary vascular resistance showed similar changes. Pulmonary capillary wedge pressure increased from 12 +/- 3 to 16 +/- 4 mm Hg (at 20 min; p < 0.05). Although tedisamil resulted in a significant heart-rate reduction, this was not associated with an improvement of hemodynamics. This may be due to increased afterload of the left and right ventricle. In these patients, tedisamil increased vascular resistance, which is unwanted in the treatment of congestive heart failure. PMID- 9869505 TI - Effect of ketanserin on central haemodynamics and coronary circulation. AB - Ketanserin is a selective 5-hydroxytryptamine (5-HT2) antagonist with vasodilator properties in the systemic and pulmonary circulation. Ketanserin also can inhibit serotonin-induced coronary artery vasoconstriction during percutaneous transluminal coronary angioplasty (PTCA). The in vivo effect of ketanserin on the coronary arteries of patients with stable angina has not previously been reported. The effects of intravenous ketanserin on cardiac haemodynamics and coronary artery diameter were measured in 10 patients with stable angina undergoing diagnostic cardiac catheterisation. Ketanserin (10 mg, i.v.) was associated with significant reductions in systemic and pulmonary arterial pressure (p < 0.05) and total systemic (SVR) and pulmonary (PVR) vascular resistance (p < 0.05). No significant change in mean coronary artery diameter or coronary artery stenotic index was evident after ketanserin. Vasodepressor responses in the systemic and pulmonary arterial circulation were observed after ketanserin injection. We assume these responses to be a direct effect of ketanserin, although non-drug-induced changes over time cannot be excluded. No significant effect on coronary artery diameter was observed, presumably because circulating serotonin levels are low in patients with stable anginal symptoms. PMID- 9869506 TI - Effect of the insertion/deletion polymorphism of the angiotensin-converting enzyme gene on response to angiotensin-converting enzyme inhibitors in patients with heart failure. AB - There is marked interindividual variation in serum and tissue angiotensin converting enzyme (ACE) levels for which the insertion (I)/deletion (D) polymorphism in intron 16 of the ACE gene is a marker. ACE inhibitors have important effects on morbidity and mortality in heart failure. The influence of this polymorphism on the response to ACE inhibitors in patients with heart failure is not known. We studied response by ACE genotype of 34 subjects in a randomised, double-blind, crossover study comparing 6 weeks of lisinopril (10 mg, o.d.) or captopril (25 mg, t.d.s.) on 24-h blood pressure (BP) profile and on renal function in patients with symptomatic heart failure [mean left ventricular ejection fraction (LVEF), 24%]. Glomerular filtration rate (GFR), 99mTc diethylenetriaminepentaacetic acid (DTPA), and ambulatory 24-h mean arterial pressure (MAP; Spacelabs 90207) were assessed at the beginning and end of treatment periods. There was a significant relation between ACE genotype and change in MAP with captopril (mm Hg; DD group, -0.5; ID, -4.7; II, -7.4; p = 0.02) but not to lisinopril (mm Hg DD, -6.0; ID, -6.6; II, -7.4; p = 0.89) in these patients. There was no significant relation between genotype and change in GFR with captopril (percentage change from baseline: DD, +7.9; ID, +13.1; II, 0.6; p = 0.45) or lisinopril (percentage change from baseline: DD, -0.1; ID, 3.0; II, -13.3; p = 0.39), but the decline in renal function tended to be greatest in II subjects. Whereas the results are not conclusive, there may be a significant interaction between ACE genotype and response to ACE inhibitors in patients with heart failure. PMID- 9869507 TI - Effects of the novel antimigraine agent, frovatriptan, on coronary and cardiac function in dogs. AB - The effects of frovatriptan (VML 251/SB-209509) on coronary artery function were investigated in isolated coronary arteries from beagle dogs. Low concentrations of frovatriptan produced contraction with -logEC50 7.55 +/- 0.08 (n = 11). The maximal observed contraction attained was 56 +/- 7% of the control 5 hydroxytryptamine (5-HT; 10 microM) response. At high concentrations of frovatriptan (>6 microM), reversal of sumatriptan (10 microM)-induced contractions was noted. In arteries precontracted with the thromboxane mimetic, U46619, frovatriptan produced a bell-shaped concentration-response relation with a maximal response at 600 nM. Concentrations of frovatriptan >2 microM produced marked reversal of tone, with full relaxation of precontracted tissues at 200 microM. In anesthetized, open-chest mongrel dogs, intravenous (n = 5) or intracoronary (n = 5) artery administration of frovatriptan (0.0001-1 mg/kg) had no consistent effect on left ventricular end-diastolic pressure, left end systolic pressure, cardiac contractility, aortic blood flow, systemic peripheral resistance, coronary blood flow, coronary vascular resistance, mean arterial blood pressure, or heart rate when compared with vehicle (n = 3). Intravenous sumatriptan produced minor effects on blood pressure and heart rate. Intracoronary artery administration of sumatriptan (0.0003 mg/kg) produced an increase in systemic peripheral resistance to 120.5 +/- 8.2% compared with vehicle (97.8 +/- 5.4%; p < 0.05). This dose of sumatriptan also produced a significant increase in coronary blood flow and decrease in coronary vascular resistance. Intravenous administration of sumatriptan produced a dose-related reduction in left ventricular diastolic pressure with a reduction to 58.3 +/- 8.3% and 41.7 +/- 25% of control values observed at 0.3 and 1 mg/kg, respectively; however, administration of sumatriptan by an intracoronary route had no effect. In a model of myocardial infarction, comparable doses of sumatriptan (1.0 mg/kg) or frovatriptan (0.1 mg/kg), in terms of their effect on carotid vascular resistance, had no significant effect on infarct size. Frovatriptan had no effect on coronary blood flow after reperfusion; however, sumatriptan produced a significant reduction in coronary blood flow for < or =3 h. These studies show that frovatriptan has the capability of relaxing coronary arteries in vitro, has no overall effect on cardiac function at rest with no effect on coronary hemodynamics after myocardial infarction, and has a profile superior to that of sumatriptan. PMID- 9869508 TI - Regulation of aortic atrial natriuretic factor and angiotensinogen in experimental hypertension. AB - We investigated the relation between atrial natriuretic factor (ANF) gene expression and the status of the renin-angiotensin system (RAS) in aortic tissue in rats made hypertensive by either aortic banding or by deoxycorticosterone acetate (DOCA)-salt administration. These experimental models of hypertension are known to have differences in terms of the status of RAS. ANF messenger RNA (mRNA) levels were measured in aortic tissue by using a newly developed quantitative competitive reverse transcription polymerase chain reaction (QC-RT-PCR) technique. Changes in the proportions of alpha1 and alpha2 isoforms of Na+K+ adenosine triphosphatase (ATPase) mRNA levels were used as indicators of aortic hypertrophy. Treatment with DOCA alone, salt alone, or DOCA-salt for 5 weeks increased aortic-weight/body-weight ratio and aortic angiotensinogen mRNA levels, but did not change alpha1 or alpha2 Na+K+-ATPase mRNA levels. Aortic ANF mRNA levels had a tendency to increase after treatment with DOCA, salt, or DOCA-salt, but this change did not reach statistical significance. Suprarenal aortic banding for 6 weeks or 12 weeks increased aortic-weight/body-weight ratio (12 weeks), decreased alpha2 Na+K+-ATPase and angiotensinogen mRNA levels, but did not affect alpha1 Na+K+-ATPase mRNA levels or ANF mRNA levels. Treatment with ramipril, an angiotensin-converting enzyme (ACE) inhibitor was carried out for 6 weeks just after aortic banding (prevention experiment) or after 6 weeks in rats that were banded for the previous 6 weeks (regression experiment). High-dose ramipril (1 mg/kg)--a treatment known to inhibit both tissue and circulating RAS--normalized aortic-weight/body-weight ratio, and also normalized alpha2 Na+K+-ATPase mRNA levels. Aortic angiotensinogen mRNA levels of banded rats treated with high-dose ramipril was higher than those of the normal control, sham operated, and banded rats. Treatment with high-dose ramipril did not affect alpha1 Na+K+-ATPase mRNA levels or ANF mRNA levels. Low-dose ramipril (10 microg/kg)--a treatment that selectively inhibits tissue RAS--normalized aortic-weight/body-weight ratio but did not normalize alpha2 Na+K+-ATPase mRNA levels (regression experiment) or angiotensinogen mRNA levels (prevention experiment) and did not change either alpha1 Na+K+-ATPase mRNA levels or ANF mRNA levels. The results suggest that, in contrast to previous findings in heart and kidney, the regulation of ANF mRNA levels in aortic tissue is largely independent of pressure load, volume load, and plasma or tissue RAS. It is suggested that any antihypertrophic actions of ANF may be mediated by the increased circulating ANF levels and its interaction with its receptor or through CNP. PMID- 9869509 TI - Role of axillary lymph node dissection after tumor downstaging. PMID- 9869510 TI - Pancreatic cancer, gene delivery, and targets: the search for specificity. PMID- 9869511 TI - Intraoperative pathologic examination of the sentinel lymph node. PMID- 9869512 TI - Role of axillary lymph node dissection after tumor downstaging with induction chemotherapy for locally advanced breast cancer. AB - BACKGROUND: Induction chemotherapy has become the standard of care for patients with locally advanced breast cancer (LABC) and currently is being evaluated in prospective clinical trials in patients with earlier-stage disease. To better gauge the role of axillary lymph node dissection in patients with LABC this study was performed to assess initial axillary status on physical and ultrasound examination, axillary tumor downstaging following induction chemotherapy, and the accuracy of physical examination compared with axillary sonography in predicting which patients will have axillary lymph node metastases found on pathologic examination. METHODS: Between 1992 and 1996, 147 consecutive patients with LABC were registered in a prospective trial of induction chemotherapy using 5 fluorouracil, doxorubicin, and cyclophosphamide. Physical and ultrasound examinations of the axilla were performed at diagnosis and after induction chemotherapy. Segmental resection with axillary lymph node dissection or modified radical mastectomy was performed, followed by postoperative chemotherapy and irradiation of the breast or chest wall and regional lymphatics. RESULTS: Following induction chemotherapy, 43 (32%) of the 133 patients with clinically positive lymph nodes on initial examination had axillary tumor downstaging as assessed by physical and ultrasound examination. The sensitivity of axillary sonography in identifying axillary metastases was significantly higher than that of physical examination (62% vs. 45%, P=.012). The specificity of physical examination (84%) was higher than that of sonography (70%), but the difference did not reach statistical significance. Among the 55 patients in whom the findings of both physical and ultrasound examination of the axilla were negative following induction chemotherapy, 29 patients (53%) were found to have axillary lymph node metastases on pathologic examination of the axillary contents. However, 28 (97%) of these patients had either 1 to 3 positive lymph nodes or only micrometastases 2 to 5 mm in diameter. CONCLUSIONS: Preoperative clinical assessment of the axilla by physical examination combined with ultrasound examination is not completely accurate in predicting metastases in patients with LABC following tumor downstaging. However, patients with negative findings on both physical and ultrasound examinations of the axilla may be potential candidates for omission of axillary dissection if the axilla will be irradiated because minimal axillary disease remains. Patients who have positive findings on preoperative physical or ultrasound examinations should receive axillary dissection to ensure local control. A prospective randomized trial of axillary dissection versus axillary radiotherapy in patients with a clinically negative axilla following induction chemotherapy is currently underway. PMID- 9869513 TI - Adenovirus-mediated wild-type p53 tumor suppressor gene therapy induces apoptosis and suppresses growth of human pancreatic cancer [seecomments]. AB - BACKGROUND: The p53 tumor suppressor gene is mutated in up to 70% of pancreatic adenocarcinomas. We determined the effect of reintroduction of the wild-type p53 gene on proliferation and apoptosis in human pancreatic cancer cells using an adenoviral vector containing the wild-type p53 tumor suppressor gene. METHODS: Transduction efficiencies of six p53-mutant pancreatic cancer cell lines (AsPC-1, BxPC-3, Capan-1, CFPAC-1, MIA PaCa-2, and PANC-1) were determined using the reporter gene construct Ad5/CMV/beta-gal. Cell proliferation was monitored using a 3H-thymidine incorporation assay, Western blot analysis for p53 expression was performed, and DNA laddering and fluorescence-activated cell sorter analysis were used to assess apoptosis. p53 gene therapy was tested in vivo in a subcutaneous tumor model. RESULTS: The cell lines varied in transduction efficiency. The MIA PaCa-2 cells had the highest transduction efficiency, with 65% of pancreatic tumor cells staining positive for beta-galactosidase (beta-gal) at a multiplicity of infection (MOI) of 50. At the same MOI, only 15% of the CFPAC-1 cells expressed the beta-gal gene. Adenovirus-mediated p53 gene transfer suppressed growth of all human pancreatic cancer cell lines in a dose-dependent manner. Western blot analysis confirmed the presence of the p53 protein product at 48 hours after infection. DNA ladders demonstrated increased chromatin degradation, and fluorescence-activated cell sorter analysis demonstrated a four-fold increase in apoptotic cells at 48 and 72 hours following infection with Ad5/CMV/p53 in the MIA PaCa-2 and PANC-1 cells. Suppression of tumor growth mediated by induction of apoptosis was observed in vivo in an established nude mouse subcutaneous tumor model following intratumoral injections of Ad5/CMV/p53. CONCLUSIONS: Introduction of the wild-type p53 gene using an adenoviral vector in pancreatic cancer with p53 mutations induces apoptosis and inhibits cell growth. These data provide preliminary support for adenoviral mediated p53 tumor suppressor gene therapy of human pancreatic cancer. PMID- 9869514 TI - Use of touch preps for intraoperative diagnosis of sentinel lymph node metastases in breast cancer. AB - BACKGROUND: Intraoperative touch prep (TP) is highly accurate for determining positive breast cancer margins and thereby reducing the need for second surgeries. It also may be useful in determining the status of the sentinel lymph node (SLN) during the initial surgical resection. The objective of this study was to test the ability of intraoperative TP to predict metastatic disease and, thus, the necessity for axillary lymph node dissection (ALND) at the time of SLN biopsy. METHODS: Fifty-five patients with invasive breast cancer were entered in the SLN biopsy protocol. The SLN was identified by gamma probe, dissected, and sent to pathology for TP and permanent sections. Level I and II ALND was then performed. Any radiolabeled LN in the lymphadenectomy specimen also was sent for TP and permanent sections. RESULTS: A total of 124 radiolabeled lymph nodes (LNs) were submitted for TP; of these, 93 (75%) were SLNs. Pathologic diagnosis by TP was correct compared with permanent sections for 99.2% of the nodes. There were no false positives. There was one (0.8%) false negative. The positive predictive value was 100% and the negative predictive value was 99%. Sensitivity was 95.7% and specificity was 100%. CONCLUSIONS: TP is a simple, quick, and accurate method for detecting metastatic disease in the SLN and, when used intraoperatively, enables the surgeon to determine whether or not an ALND is necessary at the time of the initial surgery. PMID- 9869516 TI - Prognostic value of cell proliferation (Ki-67 antigen) and nuclear DNA content in clinically resectable, distal bile duct carcinoma. AB - BACKGROUND: The aim of this study was to investigate the prognostic value of cell proliferation (Ki-67 antigen) and DNA content in patients resected for distal bile duct carcinoma (DBDC). METHODS: Formalin-fixed tumor specimens of 35 patients with resected DBDC and a long-term clinical follow-up were analyzed. MIB 1 antibody was used for Ki-67 antigen detection to determine the proportion of proliferating cells. DNA content was measured using flow cytometry. RESULTS: A significant correlation was found between a low MIB-1 index (<20%) and survival (P <.05). Of the 35 tumor specimens, 34 specimens were evaluable by flow cytometry: 22 carcinomas were diploid (65%), and 12 were aneuploid (35%). The median DNA index of aneuploid tumors was 1.36 (range, 1.09 to 1.76). No correlation of DNA-ploidy with survival time was found. CONCLUSION: In contrast to DNA-ploidy pattern, Ki-67 antigen expression showed prognostic significance in resectable DBDC. A Ki-67 positive ratio of > or =20% was associated with decreased survival time. PMID- 9869515 TI - Long-term survival in patients with ovarian metastases from colorectal carcinoma. AB - BACKGROUND: Ovarian metastases (OM) are a relatively uncommon consequence of primary colorectal carcinoma (CRC). The authors present a retrospective review of the impact of elective and therapeutic oophorectomy on the natural history of CRC. METHODS: Patients with primary CRC from January 1964 through March 1996 were reviewed. Survival from the time of OM diagnosis was estimated by the Kaplan Meier method; differences between groups were based on the log-rank test. RESULTS: A total of 155 patients were studied. Synchronous OM occurred in 90 patients (58.1%); metachronous OM occurred in 65 patients (41.9%). Estimated 5 year survival for patients with synchronous OM was 9%, versus 20% for metachronous OM (P < .0001). Resection of metastatic disease was associated with an improved 5-year survival for synchronous OM (15% vs. 0%, P=.0001) and metachronous OM (24% vs. 0%, P < .0001) if patients were disease-free postoperatively. Other clinical characteristics, including age, menopausal status, stage, and location of primary tumor, had no significant impact on survival. CONCLUSIONS: Ovarian metastases from colorectal carcinoma are associated with a poor outcome. Although there is no survival advantage associated with resection of occult microscopic disease, long-term survival is possible if patients are rendered surgically disease-free. PMID- 9869517 TI - Effects of systemic and regional chemotherapy after hepatic resection for colorectal metastases. AB - BACKGROUND: Although the survival benefit of hepatic resection for colorectal metastasis has been established, some controversy remains regarding the significance of adjuvant chemotherapy after hepatic resection. METHODS: One hundred thirty-two consecutive patients who had liver resection for colorectal metastasis at our hospital between 1980 and 1997 were studied. After curative hepatic resection, 37 patients underwent systemic chemotherapy, administered orally or intravenously, and 38 patients underwent regional chemotherapy, given intra-arterially or intraportally. Forty patients had no adjuvant chemotherapy. The chemotherapeutic agents used for oral administration were uracil and Tegafur or Tegafur alone. Mitomycin C (MMC) or 5-FU was used for IV chemotherapy. Combinations of 5-FU/leucovorin or MMC/5-FU (doxorubicin) were used for regional chemotherapy. Univariate and multivariate analyses were applied to test the significance of adjuvant chemotherapy for patient survival or disease-free survival. RESULTS: Overall 5-year survival was 42.2% (95% CL: 31.2%, 53.2%). Among the possible prognostic factors studied, univariate analysis showed a significant difference in survival based on the number of tumors and lymph node metastases in the hepatic hilum. There was a significant difference in disease free survival based on adjuvant chemotherapy and lymph node metastasis. The multivariate analysis for patient survival selected four prognostic factors (P < .05), including adjuvant chemotherapy, lymph node metastasis, disease-free interval, and tumor size. The multivariate analysis for disease-free survival selected adjuvant chemotherapy, lymph node metastasis, and disease-free interval as significant factors. The most common recurrence site was remnant liver, regardless of adjuvant chemotherapy. CONCLUSIONS: Adjuvant chemotherapy significantly improved survival and disease-free survival after hepatic resection for colorectal metastases. It did not decrease recurrence rate in the remnant liver. PMID- 9869518 TI - Clinicopathologic effects of cryotherapy on hepatic vessels and bile ducts in a porcine model. AB - BACKGROUND: The proximity of a hepatic tumor to major vessels and bile ducts limits the use of cryotherapy because of the potential damage to these structures. However, the effects of cryotherapy on major hepatic vessels and bile ducts are not well understood. METHODS: Nine pigs underwent laparotomy and intraoperative ultrasound to identify hepatic vessels larger than 5.0 mm. Cryotherapy consisting of two freeze-thaw cycles was performed, incorporating the identified vessel. In four pigs the Pringle maneuver was performed to determine the effects of partial vascular occlusion on the hepatic parenchyma and structures undergoing cryotherapy. The animals were sacrificed 30 days postoperatively, and the livers were processed for histologic examination. RESULTS: Eight of the nine livers had vessels larger than 5.0 mm incorporated into the iceball, with all vessels having evidence of infarction but remaining patent. All the livers had major bile ducts incorporated in the iceball, with eight having evidence of infarction. The Pringle maneuver had no real effect on the degree of vessel and bile duct infarction. There was no incidence of hepatic bleeding, liver fracture, bile leak, or hemobilia. CONCLUSIONS: Cryotherapy results in the infarction of major hepatic vessels and bile ducts but can be safely performed in the porcine model. Proximity of tumors to major vascular and biliary structures may not be a contraindication to the use of cryotherapy. Further studies are necessary to determine whether cryotherapy can be used in humans. PMID- 9869519 TI - Accuracy of AJCC staging for breast cancer patients undergoing re-excision for positive margins. American Joint Committee on Cancer. AB - BACKGROUND: The current AJCC protocol for breast cancer staging does not include additional tumor found at the time of re-excision in the calculation of tumor size. We hypothesize that the AJCC protocol may result in understaging and undertreatment of breast cancer patients who have additional tumor found at re excision. METHODS: In a retrospective chart review of breast cancer patients, patients with tumor present at re-excision for positive margins were placed in group 1 (n=72); patients with no tumor present at re-excision, or who underwent a single, negative margin procedure were placed in group 2 (n=147). RESULTS: Patients in group 1 had a higher risk of nodal metastases when compared to patients in group 2. Mean tumor size did not differ significantly between the subgroups. Positive re-excision was strongly associated with lymph node metastases on multivariate analysis after correction for age, grade, stage, and lymphatic invasion (odds ratio=3.13, 95% CI=1.58 6.18, P=.0011). CONCLUSIONS: Current AJCC guidelines may result in undertreatment of breast cancer patients with positive re-excisions. The presence of additional tumor at the time of re excision should be considered when determining the need for systemic therapy, and may be relevant in determining T stage. PMID- 9869520 TI - Assessment of treatment for patients with primary ductal carcinoma in situ in the breast. AB - BACKGROUND: Current mammographic technology has resulted in increased detection of ductal carcinoma in situ (DCIS). It is necessary to assess which patients presenting with DCIS are good candidates for breast conservation and which of these patients should receive adjuvant radiation. METHODS: We accrued clinical data for 124 patients with a primary diagnosis of DCIS from 1979 through 1994. Primary therapy was a mastectomy for 18 patients, and a lumpectomy for 106 patients. Only 18 of the latter group of patients received adjuvant radiotherapy. For the 88 lumpectomy-alone patients (median follow-up, 5.2 years), we evaluated the effects of clinical (age and initial presentation) and pathologic (nuclear grade, architecture, parenchymal involvement, calcifications, and measured margins) factors on recurrence of DCIS or the development of invasive breast cancer. RESULTS: Patients who underwent lumpectomy with or without adjuvant radiotherapy (median follow-up, 5.0 years) were significantly more likely to have recurrence of DCIS (P=.05) than those who underwent mastectomy (median follow-up, 6.7 years): 18% (19/106) versus 0% (0/18), respectively; lumpectomy-alone patients experienced a 19% (17/88) rate of DCIS recurrence. All recurrent DCIS was ipsilateral. For lumpectomy-alone patients, the factors associated with ipsilateral recurrence of DCIS were extent of involvement of the parenchyma (P=.01, for univariate; P=.07, for multivariate) and initial presentation (P=.05, for univariate; P=.07, for multivariate). Eleven lumpectomy-alone patients developed invasive breast cancer (6 ipsilateral, 5 contralateral); none of the 18 lumpectomy patients who received adjuvant radiation developed invasive disease. None of the factors investigated, including primary surgery and adjuvant radiotherapy, were associated with a significant effect on the development of invasive disease. CONCLUSIONS: Longer follow-up is required to determine if the benefits of either mastectomy or radiotherapy following lumpectomy persist. There is a suggestion that patients under 40 years of age or women who present with nipple discharge might be considered for either adjuvant radiotherapy following lumpectomy or a simple mastectomy. PMID- 9869521 TI - Malignant melanoma of the mucous membranes: a review of 119 cases. AB - BACKGROUND: Melanomas arising from the mucous membranes lining the respiratory, digestive, and genitourinary tracts are rare. Women are more commonly affected than are men, mainly because there is no male counterpart for vulvovaginal lesions. The mainstay of therapy is surgery, with little current use of adjuvant modalities in primary therapy. These lesions usually are advanced at initial presentation; consequently, the prognosis is poor, with 5-year survivals well below 50% in most series. METHODS: One hundred and nineteen patients with primary mucosal melanoma were reviewed. They represented 1.1% of the 10,393 melanoma patients seen at Duke University between 1970 and 1995. All data were obtained from the patients' clinic charts and computerized databases. RESULTS: There were 43 tumors arising from the head and neck region, 46 from the urogenital tract, and 30 from the anorectum. A female predominance was observed, with a female-to male ratio of 2.7:1. All but five of the patients underwent resection with curative intent. Regional or distant metastases, or both, were encountered in 36 patients at the time of presentation. In patients with head and neck and urogenital tumors, local recurrences accounted for most of the treatment failures, whereas systemic recurrences were more common with tumors arising in the anorectum. The age and gender of the patient, anatomic site of origin of the tumor, clinical stage at initial presentation, and ulceration of the primary all clearly affected prognosis. Overall, the probabilities of being alive 1, 5, and 10 years after diagnosis were 80%, 29%, and 15%, respectively. CONCLUSIONS: Widely accepted classification systems are needed so that results from separate institutions can be compared adequately. Multi-institutional trials could help in delineating standardized therapeutic protocols and in establishing the potential roles of emerging modalities in the treatment of this subtype of melanoma. PMID- 9869522 TI - Ovarian cancer-associated lymphocyte recognition of folate binding protein peptides. AB - BACKGROUND: Tumor-associated lymphocytes (TAL) isolated from ovarian cancer patients contain cytotoxic T lymphocytes (CTL) capable of recognizing specific HLA/peptide complexes on tumor cells leading to tumor cell lysis. Currently, HER2/neu, overexpressed in only 30% of breast and ovarian cancers, is the only known source of CTL-recognized peptides in epithelial cancers. Therefore, we have investigated peptides derived from folate binding protein (FBP), which is over expressed in more than 90% of ovarian cancers and in the majority of other epithelial tumors. METHODS: TAL were isolated from the malignant ascites of four consecutive HLA-A2+ ovarian cancer patients and incubated in IL-2. Initial chromium-release assays were performed within 1 week. T2 cells, incubated with peptide, were used to reconstitute T cell epitopes. The FBP sequence was interrogated for HLA-A2 binding peptides, and five were synthesized (E37-41). RESULTS: Freshly cultured, unstimulated ovarian TAL recognize peptides derived from FBP. These peptides are presented in the context of HLA-A2, and are specifically recognized in a HLA class I-restricted fashion. TAL recognition of these reconstituted T cell epitopes is concentration dependent. Furthermore, the FBP peptides are shown by cold target inhibition studies to be naturally processed and presented antigens. CONCLUSIONS: FBP peptides are recognized by freshly isolated TAL from ovarian cancer patients, suggesting in vivo expression and sensitization. Because FBP is over-expressed 20-fold in most adenocarcinomas, these peptides may be used in a widely applicable peptide-based vaccine for epithelial tumors. PMID- 9869523 TI - The risk of gastrointestinal carcinoma in familial juvenile polyposis. AB - BACKGROUND: Familial juvenile polyposis (JP) is an autosomal dominant condition in which affected individuals develop upper or lower gastrointestinal (GI) juvenile polyps, or both, and have a predisposition to cancer of the gastrointestinal tract. The risk of GI cancer has not been well defined because of the small number of these families and the lack of follow-up. The objective of this study was to determine the prevalence and age at diagnosis of GI polyposis and cancer in a large JP kindred. METHODS: Medical records were reviewed, patients were interviewed, and histories were taken. Pathology reports and slides were reviewed by our pathologists. A database was created for analysis of clinical and pathologic factors. RESULTS: This kindred contains 117 members, 29 of whom have had upper or lower GI polyps or cancer, or both. All those affected have had colonic juvenile polyps or cancer, except for two who died of advanced gastric cancer and never had colonic evaluation. Nine individuals have had both upper and lower GI polyps or cancer. Sixteen of 29 (55%) affected patients have developed gastrointestinal cancer. Eleven (38%) have had colon cancer, and six (21%) have had upper GI cancers. CONCLUSIONS: The risk of gastrointestinal malignancy in affected members of this JP kindred exceeds 50%. The high risk of GI cancer warrants frequent endoscopic screening of both affected and at-risk family members. Screening will soon be facilitated by presymptomatic genetic testing for the identification of gene carriers. PMID- 9869524 TI - Volume of resection in patients treated with breast conservation for ductal carcinoma in situ. AB - BACKGROUND: The optimal treatment of ductal carcinoma in situ (DCIS) is one of the most controversial issues in the management of breast cancer. Identification of factors that affect the risk of local recurrence is very important as the incidence of DCIS increases and the use of breast conservation becomes more widespread. Because the extent of resection may affect the relapse rate, we hypothesized that larger volumes of resection (VR) may account for the lower local recurrence rates we have previously found in elderly patients. METHODS: Between 1978 and 1990, 173 cases of histologically confirmed DCIS were treated at MSKCC with breast conservation therapy. Of these, complete VR data were available for 126 cases. The VRs thus obtained were divided into two groups, <60 cm3 and > or =60 cm3, and were evaluated for correlating factors. The patients were divided into three groups by age at diagnosis: younger than 40 years, 40 to 69 years, and 70 years or older. RESULTS: The eldest group had a significantly greater proportion of large VRs (30%) as compared to the middle group (11%) and the youngest group (9%) (P=.03, chi2). Although not statistically significant, the large VR group had a lower 6-year actuarial local recurrence rate (5.6%) than did the small VR group (21.3%) (P=.16, log-rank test). This trend was observed even though adjuvant radiotherapy was used less often in patients who had large VRs. CONCLUSION: Breast conservation surgery for DCIS in elderly patients is more likely to employ a large VR. This may explain, at least in part, the observation that elderly patients have a lower local recurrence rate. PMID- 9869525 TI - T wave alternans as a predictor of recurrent ventricular tachyarrhythmias in ICD recipients: prospective comparison with conventional risk markers. AB - INTRODUCTION: The current standard for arrhythmic risk stratification is electrophysiologic (EP) testing, which, due to its invasive nature, is limited to patients already known to be at high risk. A number of noninvasive tests, such as determination of left ventricular ejection fraction (LVEF) or heart rate variability, have been evaluated as additional risk stratifiers. Microvolt T wave alternans (TWA) is a promising new risk marker. Prospective evaluation of noninvasive risk markers in low- or moderate-risk populations requires studies involving very large numbers of patients, and in such studies, documentation of the occurrence of ventricular tachyarrhythmias is difficult. In the present study, we identified a high-risk population, recipients of an implantable cardioverter defibrillator (ICD), and prospectively compared microvolt TWA with invasive EP testing and other risk markers with respect to their ability to predict recurrence of ventricular tachyarrhythmias as documented by ICD electrograms. METHODS AND RESULTS: Ninety-five patients with a history of ventricular tachyarrhythmias undergoing implantation of an ICD underwent EP testing, assessment of TWA, as well as determination of LVEF, baroreflex sensitivity, signal-averaged ECG, analysis of 24-hour Holter monitoring, and QT dispersion from the 12-lead surface ECG. The endpoint of the study was first appropriate ICD therapy for electrogram-documented ventricular fibrillation or tachycardia during follow-up. Kaplan-Meier survival analysis revealed that TWA (P < 0.006) and LVEF (P < 0.04) were the only significant univariate risk stratifiers. EP testing was not statistically significant (P < 0.2). Multivariate Cox regression analysis revealed that TWA was the only statistically significant independent risk factor. CONCLUSIONS: Measurement of microvolt TWA compared favorably with both invasive EP testing and other currently used noninvasive risk assessment methods in predicting recurrence of ventricular tachyarrhythmias in ICD recipients. This study suggests that TWA might also be a powerful tool for risk stratification in low- or moderate-risk patients, and needs to be prospectively evaluated in such populations. PMID- 9869526 TI - Verapamil-sensitive left anterior fascicular ventricular tachycardia: results of radiofrequency ablation in six patients. AB - INTRODUCTION: Verapamil-sensitive left ventricular tachycardia (VT) with a right bundle branch block (RBBB) configuration and left-axis deviation has been demonstrated to arise from the left posterior fascicle, and can be cured by catheter ablation guided by Purkinje potentials. Verapamil-sensitive VT with an RBBB configuration and right-axis deviation is rare, and may originate in the left anterior fascicle. METHODS AND RESULTS: Six patients (five men and one woman, mean age 54+/-15 years) with a history of sustained VT with an RBBB configuration and right-axis deviation underwent electrophysiologic study and radiofrequency (RF) ablation. VT was slowed and terminated by intravenous administration of verapamil in all six patients. Left ventricular endocardial mapping during VT identified the earliest ventricular activation in the anterolateral wall of the left ventricle in all patients. RF current delivered to this site suppressed the VT in three patients (ablation at the VT exit). The fused Purkinje potential was recorded at that site, and preceded the QRS complex by 35, 30, and 20 msec, with pace mapping showing an optimal match between the paced rhythm and the clinical VT. In the remaining three patients, RF catheter ablation at the site of the earliest ventricular activation was unsuccessful. In these three patients, Purkinje potential was recorded in the diastolic phase during VT at the mid-anterior left ventricular septum. The Purkinje potential preceded the QRS during VT by 66, 56, and 63 msec, and catheter ablation at these sites was successful (ablation at the zone of slow conduction). During 19 to 46 months of follow-up (mean 32+/-9 months), one patient in the group of ablation at the VT exit had sustained VT with a left bundle branch block configuration and an inferior axis, and one patient in the group of ablation at the zone of slow conduction experienced typical idiopathic VT with an RBBB configuration and left axis deviation. CONCLUSION: Verapamil-sensitive VT with an RBBB configuration and right-axis deviation originates close to the anterior fascicle. RF catheter ablation can be performed successfully from the VT exit site or the zone of slow conduction where the Purkinje potential was recorded in the diastolic phase. PMID- 9869527 TI - Comparison of ECG variables of dispersion of ventricular repolarization with direct myocardial repolarization measurements in the human heart. AB - INTRODUCTION: QT dispersion (QTD) from the 12-lead ECG has been widely adopted as a noninvasive index of dispersion of ventricular repolarization (DVR). QTD, however, has never been validated by direct comparison with myocardial DVR in the human heart. METHODS AND RESULTS: Monophasic action potential (MAP) recordings obtained in an earlier study were retrospectively matched with 12-lead ECGs available from within 24 hours of the invasive procedure. MAPs were available from an average of 8+/-3 left endocardial sites in 4 patients with left ventricular hypertrophy (LVH) and 7 patients with normal ECGs, and 6+/-2 epicardial sites in 3 patients of each group during normal ventricular activation. Local repolarization time (RT) was determined as MAP duration at 90% repolarization plus the local activation time. Dispersion of RT was calculated as the difference between the earliest and latest RT. ECGs were digitized and analyzed with recently described interactive QTD analysis software. In addition to standard QTD (defined as QTmax-QTmin), all currently proposed ECG dispersion variables were compared and correlated with the invasive measurements of DVR. QTD exhibited a reasonable correlation with dispersion of RT (R = 0.67; P < 0.01). Several other variables designed to measure DVR exhibited a similar, but not better, correlation. Among them, the QT peak/QT end ratio in V3 (R = -0.72; P < 0.01) and averaged over all analyzable leads (R = -0.59; P < 0.01) exhibited a good correlation with dispersion of RT, which was further improved when endocardial measurements were considered alone. T area measures did not correlate with dispersion of RT, but discriminated LVH. CONCLUSION: DVR can be assessed by means of a 12-lead surface ECG. Several of the variables under study exhibit a similar accuracy in determination of true myocardial dispersion of repolarization. Variables involving the terminal part of repolarization, such as the QT peak/QT ratio, even from a single lead, may add to the determination of DVR from the human heart. PMID- 9869528 TI - Lead fixation in dogs achieved with RF energy. AB - INTRODUCTION: The purpose of this study was to test whether radiofrequency (RF) energy could be used to fixate leads to the endocardium. METHODS AND RESULTS: In six dogs we measured the dislodgment force and pacing threshold before and after RF fixation in the coronary sinus (CS) and right ventricle (RV). RF fixation was achieved with a CardioRhythm Atakr ablation unit. The dislodgment force of CS leads fixed with RF energy was 1.63+/-0.65 oz, compared with < 0.1 oz for similar leads placed in the CS of six separate dogs. In the RV, leads fixed with RF energy had a dislodgment force of 1.29+/-0.27 oz, compared with 0.48+/-0.28 oz. for urethane (P < 0.01) and 1.01+/-0.21 oz for silicone (P = 0.41) tined leads. In the CS, the pacing threshold for RF fixed leads increased significantly from 2.2+/-1.1 V (0.5 msec) before fixation to 4.2+/-1.3 V after fixation (P < 0.01), while in the RV, the pacing threshold increased from 0.41+/-0.05 V (0.5 msec) before fixation to a mean of 2.03+/-0.44 V after fixation (P < 0.01). In another group of six dogs studied for 12 weeks, 5 of 6 RF fixed CS leads remained attached, as did 8 of 10 RF fixed RV leads. For the RV leads, the mean pacing threshold was 0.90+/-0.35 V, compared with 0.53+/-0.18 V (0.5 msec) for similar tined leads (P = 0.02) and 1.2+/-0.30 V (0.5 msec) for screw leads (P = 0.18) in the RV. CONCLUSION: We conclude that RF energy can be used to attach leads to the RV and CS endocardium. While the RV pacing thresholds increased acutely, the mean chronic thresholds were not significantly different for RF fixed leads and standard tined or screw leads. PMID- 9869529 TI - Evolution of the organization of epicardial activation patterns during ventricular fibrillation. AB - INTRODUCTION: This study quantified how the organization of epicardial activation changes during the first 40 seconds of ventricular fibrillation (VF). METHODS AND RESULTS: Unipolar potentials were mapped from a 504 (24 x 21) electrode array (2 mm interelectrode spacing) on the anterior right ventricle (RV) and left ventricle (LV) epicardium. The array covered approximately 20% of the epicardial surface. In each of seven pigs, six episodes of VF were induced by premature stimulation. One-half second epochs of VF were analyzed, starting 0, 10, 20, 30, and 40 seconds post induction and using novel pattern analysis algorithms. Eight parameters were quantified: (1) the number of wavefronts; (2) the epicardial area activated by wavefronts; (3) the fraction of wavefronts arising from epicardial breakthrough or from a focus; (4) the fraction of wavefronts terminated by conduction block; (5) the multiplicity index (number of distinct activation pathways in the rhythm); (6) the repeatability index (number of times activation pathways are traversed); (7) the activation rate; and (8) the wavefront propagation velocity. The results showed that VF patterns were less organized at 10 than at 0 seconds, with more, smaller wavefronts traversing a larger variety of pathways for fewer repetitions. VF activation patterns then gradually reorganized up to 40 seconds, but by a different mechanism: the spatial size of subpatterns grew, but the dynamics otherwise appeared unchanged. During both transitions, both activation rate and propagation velocity slowed monotonically. CONCLUSION: Thus, changes in organization during VF can occur by multiple mechanisms. PMID- 9869530 TI - Radiofrequency current application on immature porcine atrial myocardium: no evidence of areas of slow conduction after 12-month follow-up. AB - INTRODUCTION: Radiofrequency current (RFC) application is a widely used procedure for treatment of supraventricular arrhythmias. The purpose of this study was to investigate late electrophysiologic sequelae of RFC lesions at immature atrial myocardium in pigs, as they have not yet been systematically investigated in vitro. METHODS AND RESULTS: RFC application (temperature guided) was performed in seven piglets (mean age 6 weeks) by a steerable 6-French electrode catheter positioned at the lateral aspect of the tricuspid valve annulus. After 12 months, hearts were removed, and lesions with surrounding tissue were isolated. The viable tissue at the border of the specimen was paced with a cycle length of 500 and 600 msec. One hundred fifty impalements were performed on each specimen using capillary microelectrodes to record action potential characteristics from the lesion's surface and the surrounding tissue. In all seven specimens, no transmembrane action potentials from the fibrotic surface of each of the lesions could be recorded. The surrounding viable tissue was sharply demarcated electrically. No areas of slow conduction were detected. Action potential characteristics as mean maximum diastolic transmembrane potential, mean action potential duration at 90% repolarization, and upstroke velocity of phase 0 of the action potential were all normal. CONCLUSION: No evidence of areas of slow conduction 12 months after RFC application at immature atrial myocardium suggests that this technique is safe regarding occurrence of late atrial tachyarrhythmias after the procedure. PMID- 9869531 TI - Spatiotemporal effects of syncytial heterogeneities on cardiac far-field excitations during monophasic and biphasic shocks. AB - INTRODUCTION: It has recently been postulated that syncytial (anatomic) heterogeneities inherent within cardiac tissue might represent a significant mechanism underlying field-induced polarization of the bulk myocardium. This simulation study examines and characterizes the spatiotemporal excitatory dynamics associated with this newly hypothesized mechanism. METHODS AND RESULTS: Two-dimensional regions of syncytially heterogeneous cardiac tissue were simulated with active membrane kinetics. Heterogeneities were manifested via random spatial variations of intracellular volume fractions over multiple length scales. Excitation thresholds were determined for uniform rectangular monophasic (M) and symmetric biphasic (B) far-field stimuli, from which strength-duration and strength-interval relationships were constructed. For regions measuring 5.4 x 5.4 mm, baseline diastolic thresholds for longitudinal (L) and transverse (T) shocks of 5-msec total duration averaged (in V/cm, n = 10) M-L = 2.87+/-0.26, M-T = 6.71+/-0.83, B-L = 3.22+/-0.25, and B-T = 7.93+/-0.51. These thresholds decreased by 15% to 25% when the region sizes were increased to 10.8 x 10.8 mm. Strength-duration relationships correlated strongly with the Weiss-Lapicque hyperbolic relationship, with rheobases and chronaxies of 2.33 V/cm and 1.15 msec for M-L stimuli, and 2.28 V/cm and 2.04 msec for B-L stimuli. Strength-interval relationships for M-L and B-L stimuli decreased monotonically with increasing coupling intervals, with similar minimum coupling intervals at absolute refractoriness. However, the B-L thresholds were substantially less sensitive to changes in coupling intervals than their M-L counterparts. CONCLUSION: This study provides strong additional support for and understanding of the syncytial heterogeneity hypothesis and its manifested properties. Furthermore, these results predict that syncytial heterogeneities of even modest proportions could represent a significant mechanism contributing to the far-field excitation process. PMID- 9869532 TI - Effect of skin electrode location on radiofrequency ablation lesions: an in vivo and a three-dimensional finite element study. AB - INTRODUCTION: OBJECTIVES: To assess the effect of skin electrode location on radiofrequency (RF) ablation lesion dimensions and energy requirements. BACKGROUND: Little is known about the effects of skin electrode location on RF ablation lesion dimensions and efficiency. METHODS AND RESULTS: Temperature controlled ablation at 60 degrees C for 60 seconds was performed in six sheep. Paired lesions were created in the lateral, anterior, posterior, and septal walls of both the ventricles. For group 1 lesions, the skin electrode was positioned directly opposite the catheter tip (optimal). For group 2 lesions, we used either the standard posterior location or an anterior location if the posterior skin electrode location was used for group 1. Group 1 lesions were 5.8+/-0.8 mm deep and 9.3+/-1.9 mm wide, compared with 4.6+/-1.0 mm deep and 7.7+/-1.9 mm wide group 2 lesions (P < or = 0.001). Group 1 lesion dimensions also had less variability. A finite element model was used to simulate temperature-controlled ablation and to study the effect of skin electrode locations on lesion dimensions, ablation efficiency, and blood heating. The optimal location was 1.6 times more efficient, and the volume of blood heated to > or = 90 degrees C was 0.005 mm3 for optimal versus 2.2 mm3 for the nonoptimal location. CONCLUSION: Optimal skin electrode placement: (1) creates deeper and larger lesions; (2) reduces lesion size variability; and (3) decreases blood heating. PMID- 9869533 TI - Cytochalasin D as excitation-contraction uncoupler for optically mapping action potentials in wedges of ventricular myocardium. AB - INTRODUCTION: Cytochalasin D in tissue bath superfusate inhibits the contraction of isolated thin trabeculae from canine right ventricle without affecting the intracellular action potential recorded with glass microelectrode. The purpose of this study was to test whether cytochalasin D could also be used to immobilize perfused wedges of ventricular muscle without affecting the action potential duration or propagation, and also to determine the optimal concentration and time duration of drug in the perfusate. METHODS AND RESULTS: Using a membrane potential sensitive dye, di-4-ANEPPS, and a high-resolution photodiode optical mapping system at a rate of 1,000 frames/sec, we recorded action potentials on the transmural surface of arterially perfused wedges of muscle from the canine left ventricular free wall. We also recorded arterial pulse pressure as a surrogate for tissue contraction. Cytochalasin D at > or = 20 micromol/L in the perfusate for > or = 6 minutes reduced the arterial pulse pressure to approximately one tenth of its initial value and significantly reduced or eliminated motion artifacts in the action potentials. A sustained concentration of 10 micromol/L cytochalasin D in the perfusate prevented contraction from recurring after the tissue was immobilized with an initial concentration of 25 micromol/L. Cytochalasin D had little effect on the action potential duration and on its transmural gradient, and did not slow the transmural velocity of excitation propagation. CONCLUSION: Cytochalasin D can be used to uncouple excitation and contraction in perfused canine cardiac muscle for the fluorescent optical mapping of action potentials without affecting action potential duration or slowing transmural propagation. PMID- 9869534 TI - Differential effects of cytochalasin D and 2,3 butanedione monoxime on isometric twitch force and transmembrane action potential in isolated ventricular muscle: implications for optical measurements of cardiac repolarization. AB - INTRODUCTION: 2,3-Butanedione monoxime (BDM) has been widely used to inhibit contraction during optical recordings of cardiac membrane voltage changes, even though it markedly abbreviates cardiac action potentials. METHODS AND RESULTS: We compared the effects of BDM and of the F-actin disrupter cytochalasin D (cyto D) on isometric twitch force and transmembrane action potentials in isolated canine right ventricular trabeculae superfused with Tyrode's solution (2 mmol/L CaCl2, 37 degrees C) and stimulated at 0.5 Hz. BDM at 10 mmol/L and cyto D at 80 micromol/L were equally effective in reducing peak isometric force to 10%+/-3% (n = 6; mean+/-SEM) and 8%+/-1% (n = 8), respectively. Neither agent significantly altered resting tension. While 10 mmol/L BDM markedly shortened the action potential duration at 90% repolarization (APD90) from 198+/-7 msec to 146+/-9 msec (P < 0.001), 80 micromol/L cyto D had no significant effects on APD90 or on any other action potential parameter. The effects of BDM on peak isometric force and APD were completely reversible after 15 minutes of washout, whereas in the cyto D group contractile force continued to be reduced (13%+/-3%) and action potential characteristics did not show significant changes from control values after a 60-minute period of superfusion with cyto D-free Tyrode's solution. CONCLUSION: We conclude that cyto D should be considered an alternative excitation-contraction uncoupler for optical mapping studies of cardiac repolarization. PMID- 9869535 TI - A fungal metabolite that eliminates motion artifacts. PMID- 9869536 TI - New diagnostic finding to assess para-Hisian pacing observed in a patient with a permanent form of junctional reciprocating tachycardia. AB - Para-Hisian pacing, a useful method to differentiate conduction over an accessory pathway from conduction over the AV node, is assessed essentially by comparing the timing of local atrial electrograms between His-bundle captured beats and His bundle noncaptured beats. We describe the case of a patient with a permanent form of junctional reciprocating tachycardia, in whom an atrial double potential was recorded only during the tachycardia at the right posterior septum. During para Hisian pacing, a morphologic change in the atrial electrogram at the posterior septum was also identified, as well as a change in the retrograde atrial sequence. Since the morphologic change of atrial electrograms during para-Hisian pacing cannot be demonstrated in a patient without an accessory pathway, this new finding could be considered a new additional diagnostic criterion suggesting the presence of an accessory pathway. PMID- 9869537 TI - The Pediatric Radiofrequency Ablation Registry's experience with Ebstein's anomaly. Pediatric Electrophysiology Society. AB - INTRODUCTION: Abnormal anatomy and complex electrophysiology in patients with Ebstein's anomaly of the tricuspid valve may confound attempts at radiofrequency ablation (RFA). METHODS AND RESULTS: Data for 65 pediatric Ebstein's patients (9.8+/-5.4 years, 4 months to 20 years; 39+/-25 kg, 5.1 to 108 kg) were obtained from the Pediatric Radiofrequency Ablation Registry. The degree of tricuspid regurgitation (DOTR) and the degree of Ebstein's anomaly were assessed with echocardiography/Doppler. Leading indications were drug refractoriness (24 [37%] of 65 patients) and life-threatening arrhythmia (14 [22%] of 65 patients). For the 65 patients, 82 typical (nondecremental) accessory pathways (APs) (62% right free wall, 34% right septal, and 4% left sided), 17 other supraventricular tachycardias (1 ectopic atrial, 7 AV reentry, 5 Mahaim, and 4 intra-atrial reentry tachycardias), and 1 ventricular mechanism were mapped. Thirty-four (52%) of 65 patients had a single AP (21 right free wall, 10 septal, and 3 left); 19 (29%) of 65 patients multiple APs; 6 (9%) of 65 patients a single AP plus a non AP mechanism; and 6 (9%) of 65 patients non-AP mechanism(s) only. RFA acute success rates and recurrence rates for right free wall, right septal, and other mechanisms were 79%/32%, 89%/29%, and 75%/27%. Mild DOTR and a body surface area (BSA) < or = 1.7 m2 independently predicted a better acute success rate. BSA < or = 1.7 m2 also predicted long-term success. CONCLUSION: In this patient subset, life-threatening arrhythmias and multiple electrophysiologic mechanisms are commonly encountered during RFA. Mild DOTR and a BSA < or = 1.7 m2 predict a higher acute success rate. While acute success rates are relatively high, recurrence is frequent. PMID- 9869538 TI - Atrial electrophysiologic remodeling: another vicious circle? AB - INTRODUCTION: With few exceptions, acquired heart disease is the result of gradual changes in the heart, progressing during several months or years. This also includes certain cardiac arrhythmias, as for instance atrial fibrillation (AF). In spite of the important role of slowly progressing pathologic processes, most of our knowledge about mechanisms of cardiac arrhythmias is based on acute experiments. Only recently, the attention also is more focused on long-term adaptation processes like cardiac memory, electrical remodeling, and tachycardia induced cardiomyopathy. In experimental animal models, it has been shown that AF induces a vicious circle of electrophysiologic and structural changes that inevitably leads to "domestication" of the arrhythmia ("AF begets AF"). In this article, the studies on AF-induced electrophysiologic and cellular remodeling are discussed. PMID- 9869539 TI - Intermittent anterograde conduction in an accessory pathway during atrial pacing: what is the mechanism? PMID- 9869540 TI - It is bundle branch reentry linked to any kind of muscular dystrophy? PMID- 9869541 TI - Adjudicating ventilator-associated pneumonia in a randomized trial of critically ill patients. AB - PURPOSE: The purpose of this study was to evaluate an adjudication strategy for diagnosing ventilator-associated pneumonia (VAP) in a randomized trial. MATERIALS AND METHODS: In a double-blind trial of sucralfate versus ranitidine, one of four pairs of adjudicators examined each case of clinically suspected VAP. Nurse and physician notes and all relevant laboratory data were allocated to each adjudication pair in groups of five patients. Each reader in the pair decided whether the patient had VAP; differences were resolved by consensus discussion. RESULTS: The overall unadjusted study odds ratio for VAP was 0.82 (P = .21) representing a trend toward less pneumonia with sucralfate compared with ranitidine. The odds ratio adjusted for adjudication pair was 0.85 (P = .27). The proportion of charts adjudicated as VAP positive among pairs ranged from 50% to 92%; crude agreement between readers in each pair varied from 50% to 82%. When adjudicators disagreed, the final consensus was split evenly between the two adjudicators' initial opinions in two pairs; in the other two pairs, the final decision reflected one dominant initial opinion. Personnel time to adjudicate all patients with a suspicion of VAP was 74 days. CONCLUSIONS: Though adjudication of outcomes such as VAP is time-consuming, consistent decision-making requires strict criteria, training, and calibration. Patients should be assigned to adjudication teams through random allocation. PMID- 9869543 TI - Role of neurosympathetic pathways in the vascular response to sepsis. AB - PURPOSE: The aim of this study was to determine the role of sympathetic neural activity in the hemodynamic adaptations to sepsis in pigs with an emphasis on circuit adaptations. A fall in resistance to venous return (RVR) was predicted in contrast to what was previously observed in sympathetically intact animals that had no change in RVR. MATERIALS AND METHODS: We anesthetized and ventilated 13 pigs and gave 5 mg/kg of indomethacin. We measured cardiac output (CO) by thermodilution and measured pulmonary arterial (PAP), pulmonary capillary wedge (Pcw), right atrial pressure (Pra), and arterial pressure (MAP). Intermittent inflation of a 50-mL balloon in the right atrium was used to transiently arrest the circulation for the measurement of mean circulatory filling pressure (MCFP). RVR was calculated from (MCFP - Pra)/CO. Animals were divided into two groups; 6 received 10 mg/kg of the ganglionic blocker, hexamethonium and norepinephrine to maintain MAP; 7 had their spinal cords cut at C-2. After baseline measurements, all animals received 10 microg/kg/h of endotoxin for 2 hours, and hemodynamic measurements were repeated. Plasma samples were obtained for measurements of immunoreactive endothelin-1 (ET-1), which was assayed by a radioimmunoassay. RESULTS: Hexamethonium had no significant effect on hemodynamics except for an increase in heart rate. After endotoxin, MAP and SVR fell, PAP rose, and CO and RVR did not change. Spinal section resulted in an increase in heart rate and small increase in PAP and MCFR After endotoxin, there was a further increase in heart rate, PAP, and MCFP with a marked fall in MAP and CO. RVR increased from 2.1 +/- 0.46 after spinal section to 3.6 +/- 54 mm x min/L (P < .05). ET-1 in the hexamethonium group (n = 2) rose from 2.21 +/- .14 to 11.5 +/- 2.1 pg/ml at 2 hours, and in the spinal group (n = 7) from 2.04 +/- 0.77 to 6.85 +/- 3.9 pg/mL at 45 minutes. CONCLUSION: Spinal section resulted in a more profound fall in blood pressure and less increase in MCFP than in previously studied animals with sympathetic nervous system intact, but there was still an increase in RVR and PAP ET-1 is a possible mediator of the increase in RVR and PAP. PMID- 9869542 TI - Use of methylene blue in patients with refractory septic shock: impact on hemodynamics and gas exchange. AB - PURPOSE: The purpose of this study was to assess the acute effects of methylene blue, an inhibitor of nitric oxide synthesis, on hemodynamics and gas exchange in patients with refractory septic shock in a prospective clinical trial at medical and surgical intensive care units in a tertiary university hospital. PATIENTS AND METHODS: Prospective, sequential study of 10 consecutive patients admitted with severe septic shock of diverse causes and unable to achieve an adequate arterial pressure despite the use of at least two vasoactive drugs. Six of them also developed acute lung injury. All received 1 mg/kg intravenous bolus of methylene blue. Hemodynamic and respiratory parameters were measured at baseline and at 30, 60, 120, and 180 minutes after the bolus injection. RESULTS: Systolic, diastolic, mean arterial blood pressure, and systemic vascular resistance increased significantly in all patients, whereas no significant changes were observed in cardiac output, oxygen consumption, or oxygen extraction ratio. Gas exchange remained unaffected in patients with acute lung injury. CONCLUSIONS: Methylene blue had an acute vasopressor effect in patients with refractory septic shock, and it was not deleterious on respiratory function. PMID- 9869544 TI - Continuous noninvasive measurement of aortic blood flow in critically ill patients with a new esophageal echo-Doppler system. AB - PURPOSE: Determination of aortic blood flow (ABF) using esophageal Doppler has been proposed as a low invasive hemodynamic monitoring method. The esophageal echo-Doppler Dynemo 3000 (Sometec Inc., Paris, France) system, recently available on the market, is an original device measuring simultaneously, and at the same anatomic level, aortic diameter, and blood flow velocity. Until now, this material has been used exclusively for peroperative monitoring. The objectives of the study were to assess the feasibility and reliability of use for continuous measurements of ABF in hemodynamically compromised intensive care unit patients; and to compare ABF values and its change induced by preload manipulation with the cardiac output (CO) values measured simultaneously by the standard thermodilution method. MATERIALS AND METHODS: Sixty simultaneous measurements of ABF and CO were performed in 22 intensive care unit patients. In 16 hypovolemic patients, Doppler and thermodilution measurements were repeated after fluid replacement. RESULTS: Applicability of the method was 84.6% (failure of the echo-Doppler method in 4 of 26 eligible patients). Coefficient of variation of echo-Doppler-derived ABF was 3.25 +/- 2.26%. Interobserver variability was 3.3 +/- 1.6%. Close linear relationship was found between ABF and CO (r = 0.92). Average ABF/CO ratio was 73 +/- 10%, but significant variation was observed after fluid replacement. CONCLUSIONS: The echo-Doppler Dynemo 3000 system allows reliable continuous measurements of ABF in intensive care unit patients, both easily and safely. PMID- 9869546 TI - Clinical trial of an ex vivo arterial blood gas monitor. AB - PURPOSE: The purpose of this study was to test the performance of a patient attached, on demand ex vivo arterial blood gas (ABG) monitor, and to compare the frequency of ABG analysis using the monitor, where the monitor was operated by intensive care unit (ICU) staff on shock trauma and neurosurgical intensive care patients for < or = 6 days, with standard clinical laboratory analysis. MATERIALS AND METHODS: The ABG monitor (SensiCath; Optical Sensors Inc., Minneapolis, MN) incorporates fiber optic pH, PCO2, PO2, and thermistor temperature sensors in a 0.3-mL sensor chamber that attaches in line with the patient's arterial pressure tubing and connects via a fiberoptic cable to a bedside instrument. The monitor and standard clinical laboratory performance were compared following an institutionally approved protocol. Adult ICU patients (n = 30) were studied for whom an arterial cannula was required, the expected ICU stay was > 72 hours, > or = 2 ABG analyses/day were anticipated, and informed consent had been obtained. Paired comparison ABG analyses and quality assurance checks were performed daily. The frequency of ABG analyses in this study, for which monitor values were used for clinical decision making, was compared with the frequency previously reported for the same ICUs, for which the monitor and laboratory results were compared but only the latter were used for clinical decision making. RESULTS: Five hundred ABG analyses, 436 over the first 72 hours, were obtained using the monitor for patient management over 3,248 patient hours (85 +/- 47 hours/patient). Monitor laboratory comparison ABG analyses (n = 258) indicated stable performance over 6 days: For pH, the range of laboratory measurements was 7.200 to 7.540, accuracy (mean difference between monitor and laboratory measurement) was +0.013, and precision (standard deviation of difference between monitor and laboratory measurements) was +/-0.031. For PCO2, range: 18 to 78.5, accuracy: -0.8, precision: +/-3.4 mm Hg. For PO2, range: 41.0 to 344.0, accuracy: +2.3, precision: +/-12.8 mm Hg. The frequency of ABG analyses obtained using the monitor (ie, 15.0 +/- 11.6 ABGs/patient/72 hours) was significantly greater than that using the clinical laboratory (ie, 8.8 +/- 4.2 ABGs/patient/72 hours) (P = .01). CONCLUSION: The ABG monitor provides performance comparable to standard clinical laboratory analysis for < or = 6 days (< or = 144 hours), consistent with ICU arterial cannula changeout schedules. More frequent ABG analyses are obtained by critical care practitioners using the monitor compared with the clinical laboratory system, suggesting that clinical decision making based on ABG data may be limited by the frequency of ABG analysis. PMID- 9869545 TI - Validation of continuous thermodilution cardiac output in critically ill patients with analysis of systematic errors. AB - PURPOSE: Bolus thermodilution cardiac output (BCO) measurements are affected by variations in injectate volume, rate, and temperature. These variations are eliminated when CO is measured by a continuous automated thermal technique (CCO). Further, CCO eliminates the need for fluid boluses, reduces contamination risk, requires no operator, and provides a continuous CO trend. We prospectively evaluated CCO versus BCO in a population of critically ill adults with low, normal, and high CO states. We sought to discern any systematic effects of temperature fluctuations or signal-to-noise-ratios (SNR) on disparities between BCO and CCO measurements and also sought to assess the relative cost effectiveness of the CCO system. MATERIALS AND METHODS: Pulmonary artery catheterizations were performed in a convenience sample of 20 patients over 6 months. BCO data were obtained using a standardized protocol. Three bolus injections of 5% dextrose were given when each CO was within 10% of the median before averaging; otherwise five boluses were given, with the high and low values eliminated before averaging. Injectates were administered randomly through the respiratory cycle and at 1-minute intervals. CCO measurements were recorded from a Vigilance monitor pre and post BCO measurements, yielding an average CCO value. Also recorded were pre- and post-core temperatures and SNR during the first CCO measurement. Cost data included estimates of operator time for BCO determinations as well as costs of Intellicath (Baxter-Edwards, Irvine, CA) pulmonary artery catheters, Vigilance (Baxter-Edwards, Irvine, CA) monitors, conventional catheters, and injectates. RESULTS: Of the 20 patients, 15 were mechanically ventilated. A total of 306 paired CO values were obtained for analysis. CCO ranged from 2.5 to 14.4 L/min and BCO from 2.4 to 13.3 L/min. Absolute differences between CCO and BCO measurements increased with increasing CO, but percentage differences did not. Of the paired values, 77% were within 1 L/min of one another. Temperature instability and SNR independently had weak correlations with CCO/BCO disparities. The Vigilance system had a slightly higher net cost than conventional BCO, although no economical value was assigned to the clinical usefulness of continuous, as opposed to intermittent, CO monitoring. CONCLUSIONS: Continuous CO is a reliable and cost-effective alternative to bolus thermodilution CO for critically ill patients in low, normal, and high CO states. PMID- 9869547 TI - Continuous cardiac output and mixed venous oxygen saturation monitoring. AB - Continuous assessment of cardiac output and SVO2 in the critically ill may be helpful in both the monitoring variations in the patient's cardiovascular state and in determining the efficacy of therapy. Commercially available continuous cardiac output (CCO) monitoring systems are based on the pulsed warm thermodilution technique. In vitro validation studies have demonstrated that this method provides higher accuracy and greater resistance to thermal noise than standard bolus thermodilution techniques. Numerous clinical studies comparing bolus with continuous thermodilution techniques have shown this technique similarly accurate to track each other and to have negligible bias between them. The comparison between continuous thermal and other cardiac output methods also demonstrates good precision of the continuous thermal technique. Accuracy of continuous oximetry monitoring using reflectance oximetry via fiberoptics has been assessed both in vitro and in vivo. Most of the studies testing agreement between continuous SVO2 measurements and pulmonary arterial blood samples measured by standard oximetry have shown good correlation. Continuous SVO2 monitoring is often used in the management of critically ill patients. The most recently designed pulmonary artery catheters are now able to simultaneously measure either SVO2 and CCO or SVO2 and right ventricular ejection fraction. This ability to view simultaneous trends of SVO2 and right ventricular performance parameters will probably allow the clinician to graphically see the impact of volume loading or inotropic therapy over time, as well as the influence of multiple factors, including right ventricular dysfunction, on SVO2. However, the cost-effectiveness of new pulmonary artery catheters application remains still questionable because no established utility or therapeutic guidelines are available. PMID- 9869548 TI - Detrimental effects of high dobutamine doses on gastric pHi determinations in a critically ill septic patient. PMID- 9869549 TI - Recollections of Professor Henry Barcroft, F.R.S. AB - Professor Henry Barcroft, MD, FRS, Emeritus Professor of Physiology in St. Thomas' Hospital Medical School in London died on 11 January 1998, aged 93. He was born in Cambridge on 18 October 1904 where his father, Joseph Barcroft, a famous physiologist, worked with Foster and Langley and subsequently was appointed to the Chair of Physiology. Henry Barcroft followed in his father's footsteps. During his career as Professor of Physiology firstly at The Queen's University of Belfast and subsequently at St. Thomas's Hospital London, he made significant studies on the nervous and humoral control of human blood vessels. His success as a research scientist stemmed partly from his ability to simplify complex phenomena in a way that permitted them to be broken into component parts and tested and partly from his technical ingenuity that permitted simple, inexpensive measurements of difficult physiological variables. Perhaps the most important factor, however, was his ability to bring out and stimulate aptitudes and enthusiasms in others. Completely unselfish himself, he gave individuals every opportunity to develop their talents, and so make themselves known to a wide circle of interested medical scientists. In many ways, his life was a guidebook for young scientists on how to make the most of their opportunities. PMID- 9869550 TI - Heart rate variability is altered following spinal cord injury. AB - Spinal cord injury (SCI) patients are known to suffer from autonomic failure as a result of their injury. The magnitude of the dysautonomia resulting from such an injury is difficult to predict or characterize and, in varying degree, it impedes the recovery of physiological homeostasis. This study is intended to investigate the effectiveness of heart rate variability (HRV) analysis as a method of quantifying and characterizing autonomic function in patients with traumatic spinal myelopathy. HRV analysis was carried out in 13 male SCI patients (six tetraplegic, seven paraplegic) and 13 age-matched, able-bodied controls. Twenty four hour ambulatory and sleep ECG tracings were obtained. Time domain, amplitude, and power spectral analyses were used to study HRV and autonomic function. Both tetraplegic (20+/-12 ms, mean+/-SD) and paraplegic (22+/-8 ms) subjects demonstrated significant loss of low frequency 24-hour HRV compared to able-bodied controls (36+/-14 ms, p < 0.05) and during sleep. This was interpreted as being consistent with predominantly sympathetic denervation uninfluenced by degree of physical activity. There were no significant differences between groups in parasympathetically mediated high frequency HRV. We conclude that HRV analysis is capable of distinguishing between SCI or able bodied humans and among tetraplegic and paraplegic patients. Patterns of altered HRV may be useful in more completely characterizing or stratifying changes in physiology associated with injury level and may have diagnostic, prognostic, or therapeutic significance. PMID- 9869551 TI - Autonomic dysfunction in chronic intestinal pseudo-obstruction. AB - Fifteen tests were used to assess adrenergic, non-vagal cholinergic, and cardiovagal functions in 11 patients with chronic intestinal pseudo-obstruction (CIP). The three aims of this study were: 1) to ascertain the presence of and spectrum of autonomic involvement; 2) to assess the level of autonomic dysfunction; and 3) to compare the results of autonomic function tests with gastrointestinal motility patterns. Gastrointestinal motility displayed a neuropathic pattern in 10 patients. Adrenergic functions were abnormal in nine patients and non-vagal cholinergic functions in 10 patients. Cardiovagal functions were abnormal in only seven patients. The autonomic dysfunction was localized mostly to the postganglionic pathways. One patient, who had a myopathic pattern and muscle degeneration on small bowel biopsy, demonstrated normal responses to autonomic function tests. The patients with neuropathic CIP demonstrated widespread, mostly postganglionic autonomic dysfunction. Neuropathic CIP can occur with or without cardiovagal involvement. PMID- 9869552 TI - Idiopathic hypovolemia: a self-perpetuating autonomic dysfunction? AB - Unexplained episodic hypertension, hypotension, or orthostatic intolerance, tachycardia, anxiety, and flushing in 21 patients were investigated for the possibility of hypovolemia by blood volume and individual plasma catecholamines (including autocrine paracrine-born dopamine), determinations baseline, in response to upright posture and catecholamines only during the episodic blood pressure swings. Blood volume was determined by Cr51 fixed to patients' hemoglobin, free norepinephrine, epinephrine, and dopamine with dopamine sulfate following sulfatase hydrolysis, radioenzymatically. The recumbent mean 27.4+/-3% (SE) blood volume decrease from predicted values accentuating to 33.5+/-4% upright was associated with normal baseline plasma free norepinephrine, epinephrine, dopamine, dopamine sulfate, plasma renin activity, and aldosterone with normal mean postural responses from all patients except a hyperresponsive compared to controls (p < 0.04), plasma renin activity increase from 0.657+/-0.1 to 4.47+/-1.8 ng/mL/hr. During the hypertensive, hypotensive, or tachycardic episodes the moderate increase of free norepinephrine and epinephrine (p < 0.04) (but not free dopamine) contrasted with an increase of dopamine sulfate from 2.5+/-0.9 to clearly pathological values of 16.8+/-8.3 ng/mL (p < 0.0003 on % increase of individual values). We conclude that the normal (but to the degree of hypovolemia inappropriately low orthostatism- and episodes-associated sympathetic arousal) is outpaced by considerable episodic dopamine sulfate surges, reflecting extraneuronal dopamine discharge. Whether this increase contributes to the increased natriuresis directly or by inhibiting aldosterone response to renin angiotensin, perpetuating hypovolemia, remains to be established. PMID- 9869553 TI - Clinical and electrophysiologic attributes as predictors of results of autonomic function tests. AB - Autonomic dysfunction is a feature of some neuropathies and not others. It has been suggested that some clinical and electrophysiologic attributes are predictable of autonomic impairment detected using laboratory testing; however, dear guidelines are unavailable. We evaluated 138 relatively unselected patients with peripheral neuropathy who underwent neurologic evaluation, electromyography (EMG), nerve conduction studies, and autonomic function tests to determine which variables were predictive of laboratory findings of autonomic failure. The variables evaluated were 1) clinical somatic neuropathic findings, 2) clinical autonomic symptoms, and 3) electrophysiologic findings. Autonomic symptoms were strongly predictive (Rs = 0.40, p < 0.001) of autonomic failure. Among the non autonomic indices, absent ankle reflexes were mildly predictive (Rs = 0.19, p = 0.022) of autonomic impairment, but all others were not (duration, clinical pattern, severity, weakness, sensory loss). Electrophysiologic changes of an axonal neuropathy predicted autonomic impairment while demyelinating neuropathy did not. We conclude that autonomic studies will most likely be abnormal in patients who have symptoms of autonomic involvement and those who have an axonal neuropathy. PMID- 9869554 TI - Neurophysiological evaluation of sexual dysfunction in patients operated for colorectal cancer. AB - Sexual dysfunction after colorectal cancer surgery may be severe and occurs in 25% to 100% of cases. Thirty-eight patients underwent colorectal resection; eight (21%) who were totally impotent and two (5%) who had ejaculatory failure were therefore studied to better understand the neurophysiological alterations related to this type of surgery. The patients were evaluated after surgery with electrophysiological testing, including examination of the sacral reflex (SR), pudendal somatosensory evoked potential (PEP), and motor evoked potential (MEP) responses. Sudomotor skin response (SSR) was also studied in a group of patients. Of the 38 patients studied, 29 showed abnormalities: six of SR, three of PEP, six of MEP, and fourteen of SSR. Only a combination of all these tests permits correct evaluation of the sexual dysfunction. PMID- 9869555 TI - Consensus statement on the diagnosis of multiple system atrophy. American Autonomic Society and American Academy of Neurology. AB - We report the results of a consensus conference on the diagnosis of multiple system atrophy (MSA). We describe the clinical features of the disease, which include four domains: autonomic failure/urinary dysfunction, parkinsonism and cerebellar ataxia, and corticospinal dysfunction. We set criteria to define the relative importance of these features. The diagnosis of possible MSA requires one criterion plus two features from separate domains. The diagnosis of probable MSA requires the criterion for autonomic failure/urinary dysfunction plus poor levodopa responsive parkinsonism or cerebellar ataxia. The diagnosis of definite MSA requires pathological confirmation. PMID- 9869557 TI - Uptake and intracellular activity of moxifloxacin in human neutrophils and tissue cultured epithelial cells. AB - The penetration by moxifloxacin of human neutrophils (polymorphonuclear leukocytes [PMN]) and tissue-cultured epithelial cells (McCoy cells) was evaluated by a fluorometric assay. At extracellular concentrations of 5 mg/liter, the cellular-to-extracellular concentration ratios (C/E) of moxifloxacin in PMN and McCoy cells were 10.9 +/- 1.0 and 8.7 +/- 1.0, respectively (20 min; 37 degrees C). The uptake of moxifloxacin by PMN was rapid, reversible, nonsaturable (at extracellular concentrations ranging from 1 to 50 microg/ml), and not affected by cell viability. The uptake of moxifloxacin was affected by external pH and the environmental temperature. The incubation of PMN in the presence of sodium fluoride, sodium cyanide, and carbonyl cyanide m-chlorophenylhydrazone significantly decreased the C/E of this agent. Neither PMN stimulation nor phagocytosis of opsonized Staphylococcus aureus significantly affected the uptake of moxifloxacin by human PMN. This agent, at concentrations of 0.5, 1, and 5 mg/liter, induced a significant reduction in the survival of intracellular S. aureus in human PMN. In summary, moxifloxacin reaches much higher intracellular concentrations within phagocytic and nonphagocytic cells than extracellular ones, remaining active inside the neutrophils. PMID- 9869558 TI - Eradication by ceftriaxone of Streptococcus pneumoniae isolates with increased resistance to penicillin in cases of acute otitis media. AB - This multicenter, noncomparative, nonrandomized study evaluated the clinical efficacy and safety of ceftriaxone for treating acute otitis media in children following clinical failure of oral antibiotic therapy. Middle-ear fluid samples were collected on day 0 and on day 3, 4, or 5 (day 3 to 5) and were used to test whether ceftriaxone therapy can eradicate Streptococcus pneumoniae isolates with increased resistance to penicillin (MIC >/= 1 mg/liter). At the first visit, on day 0, middle-ear fluid was sampled for bacteriological testing by tympanocentesis or otorrhea pus suction. Patients were administered 50 mg of ceftriaxone/kg of body weight/day, injected intramuscularly once daily, for 3 days. A second sample was collected by tympanocentesis if a pneumococcus isolate for which the MIC of penicillin was >/=1 mg/liter was detected in the day-0 sample and if the middle-ear effusion persisted on day 3 to 5. This second sample was tested for bacterial eradication. One hundred eighty-six children aged 5 months to 5 years, 10 months, with acute otitis media clinical failure were enrolled and treated in this trial. On day 10 to 12, 145 (83.8%) of the 173 patients evaluable for clinical efficacy were clinically cured. Of the 59 patients infected by pneumococci, 36 had isolates for which the MICs of penicillin were >/=1 mg/liter. Of those patients, on day 10 to 12, 32 (88.9%) were clinically cured. Middle-ear fluid samples collected by day 3 to 5 following the onset of treatment with ceftriaxone were sterile for 24 of the 27 (88.9%) patients who were infected as of day 0 by pneumococci for which the MICs of penicillin were >/=1 mg/liter and who were evaluable for bacteriological eradication. On day 10 to 12, 81.4% of S. pneumoniae-infected children and 87.5% of Haemophilus influenzae-infected children were clinically cured. No discontinuation of treatment due to adverse events, particularly due to local reactions at the injection site, were reported. Only 11 adverse events which had doubtful, probable, or possible links with the study treatment were recorded. Both the bacteriologically assessed eradication of pneumococci for which the MICs of penicillin were >/=1 mg/liter and the clinical cure rates demonstrate that ceftriaxone is of value in the management of acute otitis media unresponsive to previous oral antibiotic therapy. PMID- 9869556 TI - Antifungal peptides: novel therapeutic compounds against emerging pathogens. PMID- 9869560 TI - Development of rifapentine susceptibility tests for Mycobacterium tuberculosis. AB - Two methods for testing the susceptibility of Mycobacterium tuberculosis to rifapentine have been developed: the agar proportion method and the radiometric BACTEC technique. A critical concentration of 0.5 microg of rifapentine per ml is proposed for both methods since it provides a reliable means of distinguishing between susceptible and resistant M. tuberculosis isolates. It is recommended that two quality control M. tuberculosis strains be used at the introduction of these tests in a clinical laboratory: one that is pansusceptible (H37Rv) and one that is resistant to rifapentine. The resistant strain can be obtained from the American Type Culture Collection, where it is deposited under the number ATCC 700457. PMID- 9869559 TI - Close association between clearance of recombinant human granulocyte colony stimulating factor (G-CSF) and G-CSF receptor on neutrophils in cancer patients. AB - Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is used to counter chemotherapy-induced neutropenia. Our previous study showed an inverse correlation between serum rhG-CSF levels and the number of circulating neutrophils in cancer patients (H. Takatani, H. Soda, M. Fukuda, M. Watanabe, A. Kinoshita, T. Nakamura, and M. Oka, Antimicrob. Agents Chemother. 40:988-991, 1996). The aim of this study was to clarify the relationship between rhG-CSF clearance and G-CSF receptors on circulating neutrophils. In five cancer patients receiving chemotherapy, a bolus dose of rhG-CSF (5 microg/kg) was injected intravenously during defined phases of posttreatment neutropenia and neutrophilia. Serum rhG-CSF levels were measured by a chemiluminescence enzyme immunoassay and analyzed by moment analysis. G-CSF receptors on neutrophils were detected by flow cytometry with biotinylated rhG-CSF. rhG-CSF clearance was significantly higher at neutrophilia than at neutropenia (1,497 +/- 132 versus 995 +/- 266 ml/h; P < 0.01). The percentage of G-CSF receptor-positive neutrophils, reflecting the number of G-CSF receptors per cell, was low at neutropenia without rhG-CSF therapy (44.5% +/- 22.1%) and high at neutrophilia with rhG-CSF therapy (73. 0% +/- 11.4%; P < 0.01). rhG-CSF clearance closely correlated with the percentage of G-CSF receptor-positive neutrophils (r2 = 0.91; P < 0.0001) and neutrophil count (r2 = 0.72; P < 0.005). Our results indicate that, in cancer patients receiving chemotherapy, rhG-CSF increases the number of G-CSF receptors per cell as well as circulating neutrophil counts, resulting in modulation of its own clearance. PMID- 9869561 TI - Two pharmacodynamic models for assessing the efficacy of amoxicillin-clavulanate against experimental respiratory tract infections caused by strains of Streptococcus pneumoniae. AB - Two models of respiratory tract infection were used to investigate the pharmacodynamics of amoxicillin-clavulanate against Streptococcus pneumoniae. Eight strains of S. pneumoniae were used in a mouse model in which the animals were infected intranasally and were then treated with a range of doses and dose intervals. The time that the plasma amoxicillin concentration remained above the MIC (T>MIC) correlated well with bacterial killing, such that if T>MIC was below 20% there was no effect on bacterial numbers in the lungs. As T>MIC increased, the response, in terms of decreased bacterial load, improved and at T>MICs of greater than 35 to 40% of the dosing interval, bacteriological cure was maximal. On the basis of equivalent T>MICs, these data would suggest that in humans a dosage of 500 mg three times daily (t.i.d.) should have efficacy equal to that of a dosage of 875 mg twice daily (b.i.d.). This hypothesis was evaluated in a rat model in which amoxicillin-clavulanate was given by computer-controlled intravenous infusion to achieve concentrations that approximate the concentrations achieved in the plasma of humans following oral administration of 500/125 mg t.i.d. or 875/125 mg b.i.d. Infusions continued for 3 days and bacterial numbers in the lungs 2 h after the cessation of the infusion were significantly reduced (P < 0.01) by both treatments in strains of S. pneumoniae for which amoxicillin MICs were below 2 microg/ml. When tested against a strain of S. pneumoniae for which the amoxicillin MIC was 4 microg/ml, the simulated 500/125-mg dose was ineffective but the 875/125-mg dose demonstrated a small but significant (P < 0. 01) reduction in bacterial numbers. These data confirm the findings in the mouse and indicate that amoxicillin-clavulanate administered at 875/125 mg b.i.d. would be as effective clinically as amoxicillin-clavulanate administered at 500/125 mg t.i.d. PMID- 9869562 TI - Efficacy of high-dose amoxicillin-clavulanate against experimental respiratory tract infections caused by strains of Streptococcus pneumoniae. AB - The purpose of the present investigation was to determine if the efficacy of amoxicillin-clavulanate against penicillin-resistant Streptococcus pneumoniae could be improved by increasing the pediatric amoxicillin unit dose (90 versus 45 mg/kg of body weight/day) while maintaining the clavulanate unit dose at 6.4 mg/kg/day. A rat pneumonia model was used. In that model approximately 6 log10 CFU of one of four strains of S. pneumoniae (amoxicillin MICs, 2 microg/ml [one strain], 4 microg/ml [two strains], and 8 microg/ml [one strain]) were instilled into the bronchi of rats. Amoxicillin-clavulanate was given by computer controlled intravenous infusion to approximate the concentrations achieved in the plasma of children following the administration of oral doses of 45/6.4 mg/kg/day or 90/6.4 mg/kg/g/day divided every 12 h or saline as a control for a total of 3 days. Infusions continued for 3 days, and 2 h after the cessation of infusion, bacterial numbers in the lungs were significantly reduced by the 90/6.4-mg/kg/day equivalent dosage for strains for which amoxicillin MICs were 2 or 4 microg/ml. The 45/6.4-mg/kg/day equivalent dosage was fully effective only against the strain for which the amoxicillin MIC was 2 microg/ml and had marginal efficacy against one of the two strains for which amoxicillin MICs were 4 microg/ml. The bacterial load for the strain for which the amoxicillin MIC was 8 microg/ml was not reduced with either dosage. These data demonstrate that regimens which achieved concentrations in plasma above the MIC for at least 34% of a 24-h dosing period resulted in significant reductions in the number of viable bacteria, indicating that the efficacy of amoxicillin-clavulanate can be extended to include efficacy against less susceptible strains of S. pneumoniae by increasing the amoxicillin dose. PMID- 9869563 TI - Vancomycin-dependent Enterococcus faecalis clinical isolates and revertant mutants. AB - Three vancomycin-dependent clinical isolates of Enterococcus faecalis of the VanB type were studied by determining (i) the sequence of the ddl gene encoding the host D-Ala:D-Ala ligase and the vanSB-vanRB genes specifying the two-component regulatory system that activates transcription of the vanB operon, (ii) the level of expression of resistance genes by using DD-dipeptidase activity as a reporter, and (iii) the proportions of the peptidoglycan precursors synthesized. Each strain had a mutation in ddl leading to an amino acid substitution (D295 to V; T316 to I) or deletion (DAK251-253 to E) at invariant positions in D-Ala:D-Ala, D Ala:D-Lac, and D-Ala:D-Ser ligases. These mutations resulted in impaired host D Ala:D-Ala ligases since only precursors terminating in D-Ala-D-Lac were synthesized under vancomycin-inducing conditions. Two types of vancomycin independent revertants of one isolate were obtained in vitro after growth in the absence of vancomycin: (i) vancomycin-resistant, teicoplanin-susceptible mutants had a 6-bp insertion in the host ddl gene, causing the E251-to-EYK change that restored D-Ala:D-Ala ligase activity, (ii) constitutive vancomycin-resistant, teicoplanin-resistant mutants had substitutions (S232 to F or E247 to K) in the vicinity of the autophosphorylation site of the VanSB sensor and produced exclusively precursors ending in D-Ala-D-Lac. Vancomycin- and teicoplanin dependent mutants obtained by growth in the presence of teicoplanin had an 18-bp deletion in VanSB, affecting residues 402 to 407 and overlapping the G2 ATP binding domain. The rapid emergence of vancomycin-independent revertants in vitro suggests that interruption of vancomycin therapy may not be sufficient to cure patients infected with vancomycin-dependent enterococci. PMID- 9869564 TI - Erythromycin resistance genes in group A streptococci in Finland. The Finnish Study Group for Antimicrobial Resistance. AB - Streptococcus pyogenes isolates (group A streptococcus) of different erythromycin resistance phenotypes were collected from all over Finland in 1994 and 1995 and studied; they were evaluated for their susceptibilities to 14 antimicrobial agents (396 isolates) and the presence of different erythromycin resistance genes (45 isolates). The erythromycin-resistant isolates with the macrolide-resistant but lincosamide- and streptogramin B-susceptible phenotype (M phenotype) were further studied for their plasmid contents and the transferability of resistance genes. Resistance to antimicrobial agents other than macrolides, clindamycin, tetracycline, and chloramphenicol was not found. When compared to our previous study performed in 1990, the rate of resistance to tetracycline increased from 10 to 93% among isolates with the inducible resistance (IR) phenotype of macrolide, lincosamide, and streptogramin B (MLSB) resistance. Tetracycline resistance was also found among 75% of the MLSB-resistant isolates with the constitutive resistance (CR) phenotype. Resistance to chloramphenicol was found for the first time in S. pyogenes in Finland; 3% of the isolates with the IR phenotype were resistant. All the chloramphenicol-resistant isolates were also resistant to tetracycline. Detection of erythromycin resistance genes by PCR indicated that, with the exception of one isolate with the CR phenotype, all M-phenotype isolates had the macrolide efflux (mefA) gene and all the MLSB-resistant isolates had the erythromycin resistance methylase (ermTR) gene; the isolate with the CR phenotype contained the ermB gene. No plasmid DNA could be isolated from the M-phenotype isolates, but the mefA gene was transferred by conjugation. PMID- 9869565 TI - Evaluation of a novel, anti-herpes simplex virus compound, acyclovir elaidate (P 4010), in the female guinea pig model of genital herpes. AB - The antiviral effect of acyclovir elaidate in the female guinea pig model of genital herpes was investigated in a series of experiments. The antiherpesvirus effects of this novel compound, 9-(2'-[trans-9" octadecenoyloxyl]ethoxymethyl)guanine (code no. P-4010), were studied in both primary and recurrent genital herpes in the female guinea pig, following oral gavage or intraperitoneal injection, with different formulations of the compound, and in comparison with acyclovir (ACV) or penciclovir (PCV). The results indicate that compound P-4010 has a greater capability than either ACV or PCV in reducing the clinical symptoms of primary genital herpes induced following the inoculation of herpes simplex virus type 2 (HSV-2) intravaginally into guinea pigs. In addition, the administration of P-4010 twice daily over a 10-day period by the intraperitoneal route (15 to 40 mg/kg of body weight/day) or by oral gavage (50 to 200 mg/kg/day), commencing 4 h subsequent to intravaginal HSV-2 infection, resulted in a degree of reduction in the incidence and severity of spontaneous, recurrent genital herpes in these animals. The findings are discussed in the light of the value and relevance of the female guinea pig model of genital herpes for the assessment of anti-herpes simplex virus compounds. PMID- 9869566 TI - Type II topoisomerase mutations in ciprofloxacin-resistant strains of Pseudomonas aeruginosa. AB - We determined the sequences of the quinolone resistance-determining regions of gyrA, gyrB, and parC genes for 30 clinical strains of Pseudomonas aeruginosa resistant to ciprofloxacin that were previously complemented by wild-type gyrA and gyrB plasmid-borne alleles and studied for their coresistance to imipenem (E. Cambau, E. Perani, C. Dib, C. Petinon, J. Trias, and V. Jarlier, Antimicrob. Agents Chemother. 39:2248-2252, 1995). In the present study, we found mutations in type II topoisomerase genes for all strains. Twenty-eight strains had a missense mutation in gyrA (codon 83 or 87). Ten of them had an additional mutation in parC (codon 80 or 84), including a novel mutation of Ser-80 to Trp, but all were fully complemented by a plasmid-borne wild-type gyrA allele. The remaining two strains harbored the first gyrB mutation described in P. aeruginosa, leading to the substitution of phenylalanine for serine 464. The strains which had two mutations in type II topoisomerase genes (i.e., gyrA and parC) were significantly more resistant to fluoroquinolones than those with a single mutation in gyrA or gyrB (geometric mean MICs of ciprofloxacin, 39.4 versus 10.9 microg/ml, P < 0.01; geometric mean MICs of sparfloxacin, 64.0 versus 22.6, P < 0. 01). No mutant with a parC mutation alone was observed, which favors DNA gyrase being the primary target for fluoroquinolones. These results demonstrate that gyrA mutations are the major mechanism of resistance to fluoroquinolones for clinical strains of P. aeruginosa and that additional mutations in parC lead to a higher level of quinolone resistance. PMID- 9869567 TI - Therapeutic effect of clarithromycin on a transplanted tumor in rats. AB - The therapeutic antitumor effect of clarithromycin (CAM) was examined with the 13762NF mammary adenocarcinoma and F-344 rat system. When CAM treatment at a dosage of 2 mg/kg of body weight orally for 21 days was commenced after inoculation of the tumor, no significant decrease in death rate was observed, although the loss in body weight was less than that in the untreated group. When tumor-bearing (TB) rats were treated with CAM in combination with carboplatin or cyclophosphamide, a significant decrease in the death rate was obtained, although neither treatment alone proved to be effective. A beneficial effect was also observed when CAM treatment was combined with surgical treatment. CAM showed no direct cytotoxicity to this tumor in vitro according to the MTT (3-[4, 5 dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Spleen cells obtained from TB rats receiving CAM treatment showed a stronger tumor neutralizing activity than those from rats which had not received CAM treatment (Winn assay). Enhanced induction of cytotoxic cells to allogeneic tumor was also observed in rats immunized with allogeneic tumor cells together with CAM treatment (51Cr release assay). The 13762NF tumor produces transforming growth factor-beta (TGF-beta), tumor necrosis factor alpha, and matrix metalloproteinase 9, and treatment of tumor cells with CAM in vitro for 24 h significantly inhibited the expression of the genes coding for these proteins (reverse transcription-PCR). Levels of expression of the TGF-beta and interleukin-6 genes of spleen cells obtained from CAM-treated TB rats were both significantly lower than those of spleen cells from CAM-untreated TB rats. This study suggests that CAM has biological response modifier activities resulting in a beneficial therapeutic antitumor effect and might be useful for the treatment of human cancers. PMID- 9869568 TI - Efficacy of new 5-nitroimidazoles against metronidazole-susceptible and resistant Giardia, Trichomonas, and Entamoeba spp. AB - The efficacies of 12 5-nitroimidazole compounds and 1 previously described lactam substituted nitroimidazole with antiparasitic activity, synthesized via SRN1 and subsequent reactions, were assayed against the protozoan parasites Giardia duodenalis, Trichomonas vaginalis, and Entamoeba histolytica. Two metronidazole sensitive lines and two metronidazole-resistant lines of Giardia and one line each of metronidazole-sensitive and -resistant Trichomonas were tested. All except one of the compounds were as effective or more effective than metronidazole against Giardia and Trichomonas, but none was as effective overall as the previously described 2-lactam-substituted 5-nitroimidazole. None of the compounds was markedly more effective than metronidazole against Entamoeba. Significant cross-resistance between most of the drugs tested and metronidazole was evident among metronidazole-resistant lines of Giardia and Trichomonas. However, some drugs were lethal to metronidazole-resistant Giardia and had minimum lethal concentrations similar to that of metronidazole for drug susceptible parasites. This study emphasizes the potential in developing new nitroimidazole drugs which are more effective than metronidazole and which may prove to be useful clinical alternatives to metronidazole. PMID- 9869569 TI - Efficacy of trovafloxacin in treatment of experimental staphylococcal or streptococcal endocarditis. AB - The efficacy of trovafloxacin against Staphylococcus aureus and viridans group streptococci was investigated in vitro and in an experimental model of endocarditis. The MICs at which trovafloxacin and ciprofloxacin inhibited 90% of clinical isolates of such bacteria (MIC90s) were (i) 0.03 and 2 mg/liter, respectively, for 30 ciprofloxacin-susceptible S. aureus isolates, (ii) 32 and 128 mg/liter, respectively, for 20 ciprofloxacin-resistant S. aureus isolates, and (iii) 0.25 and 8 mg/liter, respectively, for 28 viridans group streptococci. Rats with aortic vegetations were infected with either of two ciprofloxacin susceptible but methicillin-resistant S. aureus strains (strains COL and P8), one penicillin-susceptible Streptococcus sanguis strain, or one penicillin-resistant Streptococcus mitis strain. Rats were treated for 3 or 5 days with doses that resulted in kinetics that simulated those achieved in humans with trovafloxacin (200 mg orally once a day), ciprofloxacin (750 mg orally twice a day), vancomycin (1 g intravenously twice a day), or ceftriaxone (2 g intravenously once a day). Against the staphylococci, the activities of both trovafloxacin and ciprofloxacin were equivalent to that of vancomycin, and treatment of endocarditis with these drugs was successful (P < 0.05). However, ciprofloxacin selected for resistant derivatives in vitro and in vivo, whereas trovafloxacin was 10 to 100 times less prone than ciprofloxacin to select for resistance in vitro and did not select for resistance in vivo. Against the two streptococcal isolates, trovafloxacin significantly (P < 0.05) decreased bacterial counts in the vegetations but was less effective than the control drug, ceftriaxone. Thus, a simulated oral dose of trovafloxacin (200 mg per day) was effective against ciprofloxacin-susceptible staphylococci and was less likely than ciprofloxacin to select for resistance. The simulated oral dose of trovafloxacin also had some activity against streptococcal endocarditis, but optimal treatment of infections caused by such organisms might require higher doses of the drug. PMID- 9869570 TI - Moxifloxacin (BAY12-8039), a new 8-methoxyquinolone, is active in a mouse model of tuberculosis. AB - Moxifloxacin (BAY12-8039) is a new 8-methoxyquinolone shown to be active against Mycobacterium tuberculosis in vitro. We tested moxifloxacin for activity in mice against M. tuberculosis CSU93, a highly virulent, recently isolated clinical strain. The MIC of moxifloxacin for the CSU93 strain was 0.25 microg/ml. The serum moxifloxacin concentration after oral administration in mice peaked within 0.25 h, reaching 7.8 microg/ml with doses of 100 mg/kg of body weight; the maximum concentration and the analysis of the area under the concentration-time curve revealed dose dependency. When mice were infected with a sublethal inoculum of mycobacteria and then treated with moxifloxacin at 100 mg/kg per day for 8 weeks, the log10 CFU counts in the organs of treated mice were significantly lower than those for the control group (0.6 +/- 0.2 versus 5.6 +/- 0. 3 in the lungs and 1.5 +/- 0.7 versus 4.9 +/- 0.5 in the spleens, respectively; P < 0.001 in both organs). The effectiveness of moxifloxacin monotherapy was comparable to that seen in mice receiving isoniazid alone. Combination therapy with moxifloxacin plus isoniazid was superior to that with moxifloxacin or with isoniazid alone in reducing bacillary counts in the organs studied. Using a sensitive broth-passage subculture method, we demonstrated that 8 weeks of treatment with moxifloxacin (100 mg/kg per day) or with moxifloxacin plus isoniazid (100 mg/kg and 25 mg/kg, respectively, per day) sterilized the lungs in seven of eight and in eight of eight mice, respectively. Among surviving bacilli isolated from animals infected with a high-titer inoculum and treated for 7 weeks with low-dose moxifloxacin (20 mg/kg per day), breakthrough resistance to moxifloxacin was not observed. These results indicate that moxifloxacin is highly effective in reducing M. tuberculosis infection in mice and has activity comparable to that of isoniazid. Combination therapy with moxifloxacin and isoniazid was highly effective, suggesting that moxifloxacin may be useful in multiple-drug regimens for human tuberculosis. PMID- 9869571 TI - In vitro activity of the ketolide HMR 3647 (RU 6647) for Legionella spp., its pharmacokinetics in guinea pigs, and use of the drug to treat guinea pigs with Legionella pneumophila pneumonia. AB - The activities of HMR 3647, HMR 3004, erythromycin, clarithromycin, and levofloxacin for 97 Legionella spp. isolates were determined by microbroth dilution susceptibility testing. Growth inhibition of two Legionella pneumophila strains grown in guinea pig alveolar macrophages was also determined. The concentrations required to inhibit 50% of strains tested were 0.06, 0.02, 0.25, 0.03, and 0.02 microg/ml for HMR 3647, HMR 3004, erythromycin, clarithromycin, and levofloxacin, respectively. BYEalpha broth did not significantly inhibit the activities of the drugs tested, as judged by the susceptibility of the control Staphylococcus aureus strain; however, when Escherichia coli was used as the test strain, levofloxacin activity tested in BYEalpha broth was fourfold lower. HMR 3647, HMR 3004, erythromycin, and clarithromycin (0.25 and 1 microg/ml) reduced bacterial counts of two L. pneumophila strains grown in guinea pig alveolar macrophages by 0.5 to 1 log10, but regrowth occurred over a 2-day period. HMR 3647, erythromycin, and clarithromycin appeared to have equivalent intracellular activities which were solely static in nature. HMR 3004 was more active than all drugs tested except levofloxacin. In contrast, levofloxacin (1 microg/ml) was bactericidal against intracellular L. pneumophila and significantly more active than the other drugs tested. Therapy studies with HMR 3647 and erythromycin were performed in guinea pigs with L. pneumophila pneumonia. When HMR 3647 was given (10 mg/kg of body weight) by the intraperitoneal route to infected guinea pigs, mean peak plasma levels were 1.4 microg/ml at 0.5 h and 1.0 microg/ml at 1 h postinjection. The terminal half-life phase of elimination from plasma was 1.4 h. All 16 L. pneumophila-infected guinea pigs treated with HMR 3647 (10 mg/kg/dose given intraperitoneally once daily) for 5 days survived for 9 days after antimicrobial therapy, as did all 16 guinea pigs treated with the same dose of HMR 3647 given twice daily. Fourteen of 16 erythromycin-treated (30 mg/kg/dose given intraperitoneally twice daily) animals survived, whereas 0 of 12 animals treated with saline survived. HMR 3647 is effective against L. pneumophila in vitro, in infected macrophages, and in a guinea pig model of Legionnaires' disease. HMR 3647 given once daily should be evaluated as a treatment for Legionnaires' disease in humans. PMID- 9869572 TI - Effect of probenecid on the renal excretion mechanism of a new carbapenem, DA 1131, in rats and rabbits. AB - The effects of probenecid, an anion transport inhibitor, on the renal excretion mechanism of a new anionic carbapenem, DA-1131, were investigated after a 1-min intravenous infusion of DA-1131 at 100 mg/kg of body weight to rabbits and 50 mg/kg to rats with or without probenecid at 50 mg/kg for both species. In control rabbits, the renal clearance (CLR) of DA-1131 and the glomerular filtration rate based on creatinine clearance (CLCR) were 6.14 +/- 2.09 and 2.26 +/- 0.589 ml/min/kg, respectively. When considering the less than 10% plasma protein binding of DA-1131 in rabbits, renal tubular secretion of DA-1131 was observed in rabbits. The CLR of DA-1131 (3. 87 +/- 0.543 ml/min/kg) decreased significantly with treatment with probenecid in rabbits, indicating that the renal tubular secretion of DA-1131 was inhibited by probenecid. However, in control rats, the CLR of DA-1131 (5.80 +/- 1.94 ml/min/kg) was comparable to the CLCR (4.29 +/- 1.64 ml/min/kg), indicating that DA-1131 was mainly excreted by glomerular filtration in rats. Therefore, it could be expected that the CLR of DA-1131 could not be affected by treatment with probenecid in rats; this was proved by a similar CLR of DA-1131 with treatment with (6.93 +/- 0.675 ml/min/kg) or without (5.80 +/- 1.94 ml/min/kg) probenecid. Therefore, the renal secretion of DA-1131 is a factor in rabbits but is not a factor in rats. PMID- 9869573 TI - Acetate-mediated growth inhibition in sterol 14alpha-demethylation-deficient cells of Candida albicans. AB - Candida albicans is a fungus thought to be viable in the presence of a deficiency in sterol 14alpha-demethylation. We showed in a strain of this species that the deficiency, caused either by a mutation or by an azole antifungal agent, made the cells susceptible to growth inhibition by acetate included in the culture medium. Studies with a mutant demonstrated that the inhibition was complete at a sodium acetate concentration of 0.24 M (20 g/liter) and was evident even at a pH of 8, the latter result indicating the involvement of acetate ions rather than the undissociated form of acetic acid. In fluconazole-treated cells, sterol profiles determined by thin-layer chromatography revealed that the minimum sterol 14alpha demethylation-inhibitory concentrations (MDICs) of the drug, thought to be the most important parameter for clinical purposes, were practically identical in the media with and without 0.24 M acetate and were equivalent to the MIC in the acetate-supplemented medium. The acetate-mediated growth inhibition of azole treated cells was confirmed with two additional strains of C. albicans and four different agents, suggesting the possibility of generalization. From these results, it was surmised that the acetate-containing medium may find use in azole susceptibility testing, for which there is currently no method capable of measuring MDICs directly for those fungi whose viability is not lost as a result of sterol 14alpha-demethylation deficiency. Additionally, the acetate supplemented agar medium was found to be useful in detecting reversions from sterol 14alpha-demethylation deficiency to proficiency. PMID- 9869574 TI - Mixture-based heterocyclic combinatorial positional scanning libraries: discovery of bicyclic guanidines having potent antifungal activities against Candida albicans and Cryptococcus neoformans. AB - A mixture-based synthetic combinatorial library of more than 100,000 bicyclic guanidines was generated in a positional scanning format and assayed for activity against Candida albicans. Potent individual bicyclic guanidines were directly identified following the screening of the library. Time-kill curve studies indicated bactericidal activities for the individual bicyclic guanidines. These compounds also showed potent activity against Cryptococcus neoformans. These studies demonstrate the value of using mixture-based combinatorial positional scanning libraries made up of heterocyclic compounds for the rapid identification of novel classes of antifungal compounds. PMID- 9869575 TI - Activity and diffusion of LY333328 in experimental endocarditis due to vancomycin resistant Enterococcus faecalis. AB - The activity of LY333328 against Enterococcus faecalis JH2-2, which is susceptible to glycopeptides, and against its transconjugants E. faecalis BM4281 and BM4316, with VanB and VanA phenotypes, respectively, was investigated. LY333328 was active in vitro against the three strains, for which MICs were 2 microg/ml on agar and 0.25 microg/ml in broth. LY333328 was bactericidal in broth against E. faecalis JH2-2 and BM4281 at a concentration of 8 microg/ml and against BM4316 at a concentration of 30 microg/ml. The protein binding of LY333328 to rabbit serum was >99%, and the bactericidal activity of LY333328 in broth was reduced when it was tested in the presence of 90% rabbit serum. Autoradiographic studies performed in rabbits with enterococcal endocarditis showed that 14[C]LY333328 was distributed heterogeneously throughout cardiac vegetations. In rabbits with aortic endocarditis, a regimen of 20 mg of LY333328 per kg of body weight administered intramuscularly twice a day for 5 days after a loading dose of 40 mg/kg was active against the three strains in vivo (P < 0.01), whereas vancomycin was not active against the VanB-type strain and teicoplanin was not active against the VanA-type strain. We conclude that the activity of LY333328 is not significantly modified by acquired resistance to glycopeptides in E. faecalis either in vitro or in experimental endocarditis. PMID- 9869576 TI - Population pharmacokinetics of nevirapine, zidovudine, and didanosine in human immunodeficiency virus-infected patients. The National Institute of Allergy and Infectious Diseases AIDS Clinical Trials Group Protocol 241 Investigators. AB - The population pharmacokinetics of nevirapine (NVP), zidovudine (ZDV), and didanosine (ddI) were evaluated in a total of 175 patients infected with human immunodeficiency virus randomized to receive either a double combination of ZDV plus ddI or a triple combination of NVP plus ZDV plus ddI as a substudy of the AIDS Clinical Trials Group Protocol 241. Levels (approximating 3.5 determinations/patient) of the three drugs in plasma were measured during 44 of a total 48 weeks of study treatment, and a set of potential covariates was available for nonlinear mixed-effect modeling analysis. A one-compartment model with zero-order input and first-order elimination was fitted to the NVP data. Individual oral clearance (CL) and volume of distribution (V) averaged 0.0533 liters/h/kg of body weight and 1.17 liters/kg, respectively. Gender was the only covariate which significantly correlated with the CL of NVP. ZDV and ddI data were described by a two-compartment model with zero-order input and first-order elimination. Individual mean oral CL, VSS (volume of distribution at steady state), and V of ZDV were 1.84 liters/h/kg and 6.68 and 2.67 liters/kg, respectively, with body weight and age as correlates of CL and body weight as a correlate of VSS. The average individual oral CL, VSS, and V of ddI were 1.64 liters/h/kg and 3.56 and 2.74 liters/kg, respectively, with body weight as a significant correlate of both CL and VSS. The relative bioavailability (F) of ZDV and ddI in the triple combination compared to that in the double combination was also evaluated. No significant effects of the combination regimens on the F of ddI were detected (FTRIPLE = 1.05 and FDOUBLE = 1 by definition), but the F of ZDV was markedly reduced by the triple combination, being only 67.7% of that of the double combination. Large (>50%) intraindividual variability was associated with both ZDV and ddI pharmacokinetics. Individual cumulative area under the plasma drug level-time curve of the three drugs was calculated for the entire study period as a measure of drug exposure based on the individual data and the final-model estimates of structural and statistical parameters. PMID- 9869577 TI - Evaluation of several dosing regimens of cefepime, with various simulations of renal function, against clinical isolates of Pseudomonas aeruginosa in a pharmacodynamic infection model. AB - The objectives of this study were as follows: (i) to examine the killing activity of 2-g doses of cefepime against two clinical isolates (mucoid and nonmucoid) of Pseudomonas aeruginosa in a pharmacodynamic in vitro infection model, (ii) to compare the percentage of time above the MIC (T > MIC) for each of the regimens against P. aeruginosa, and (iii) to evaluate the area under the bactericidal curve for each regimen. Cefepime was administered at intervals of 8, 12, and 24 h with and without tobramycin, and two different levels of renal function were simulated: normal (creatinine clearance [CLCR] = 90 ml/min) and decreased (CRCL = 60 ml/min). Also, the killing activity of cefepime with and without tobramycin was compared to the killing activity of ceftazidime (2 g every 8 h) with and without tobramycin. The T > MIC was 100% in the central chamber except for the regimen in which cefepime was administered every 12 h and the CLCR was 90 ml/min, which provided concentrations above the MIC for 92% of the dosing interval against the C31 (mucoid; MIC of cefepime, 4 microg/ml) isolate and for 75% of the interval against the C34 (nonmucoid; MIC of cefepime, 8 microg/ml) isolate. All cefepime and ceftazidime monotherapy simulations resulted in 99.9% killing of the nonmucoid isolate within 4 to 8 h and within 4 to 6 h, respectively. Against the mucoid isolate, 99.9% killing was achieved only with combination therapy. The results of this study indicate that cefepime dosed at 2 g every 12 h under conditions of normal renal function and every 24 h with decreased creatinine clearance (60 ml/min) is effective both as monotherapy and in combination therapy against a nonmucoid strain of P. aeruginosa. With cefepime MICs of 4 and 8 microg/ml, the single-agent regimens provided T > MIC values in the central chamber for 92 and >/=75% of the dosing interval against the mucoid and nonmucoid isolates, respectively. Cefepime dosed at 2 g every 12 h, with a creatinine clearance of 90 ml/min, and every 24 h, with a creatinine clearance of 60 ml/min, resulted in killing activity equivalent to that of ceftazidime dosed at 2 g every 8 h. None of the monotherapies provided adequate killing of the mucoid strain of P. aeruginosa despite drug concentrations being above the MIC for >/=92% of all dosing intervals. Finally, combination therapy with tobramycin and either cefepime or ceftazidime enhanced the killing of both the mucoid and nonmucoid P. aeruginosa isolates. PMID- 9869578 TI - Limited-sampling strategy models for itraconazole and hydroxy-itraconazole based on data from a bioequivalence study. AB - The extensive interindividual variability in oral bioavailability of itraconazole prompted an assessment of the bioequivalence of two formulations marketed in Brazil, namely, Sporanox (reference) and Traconal (test). Eighteen healthy volunteers received single 200-mg oral doses of each formulation at 2-week intervals in a randomized, crossover protocol. The concentrations of itraconazole and hydroxy-itraconazole in plasma were measured by high-performance liquid chromatography, and the datum points (n = 396) were subsequently used to develop limited-sampling strategy models for estimation of the areas under the curve (AUCs) for both compounds. The 90% confidence intervals for individual percent ratios (test/reference formulations) of the maximum concentration of drug in serum, the AUC from 0 to 48 h and the AUC from time zero to infinity (AUC0 infinity) for itraconazole and hydoxy-itraconazole were below the range of 80 to 125%, suggesting that these formulations are not bioequivalent. Linear regression analysis of the AUC0-infinity against time and a "jackknife" validation procedure revealed that models based on three sampling times accurately predict (R2, >0.98; bias, <3%; precision, 3 to 7%) the AUC0-infinity for each of the four formulation compound pairs tested. Increasing the number of sampling points to more than three adds little to the accuracy of the estimates of AUC0-infinity. The three point models developed for the reference formulation were validated retrospectively and were found to predict within 2% the AUC0-infinity reported in previous studies performed under similar protocols. In conclusion, the data in this study indicate (i) that the tested formulations are not bioequivalent when single doses are compared and (ii) that limited-sampling strategy models based on three points predict accurately the AUC0-infinitys for itraconazole and hydroxy itraconazole and could be a valuable tool in pharmacokinetic and bioequivalence studies of single oral doses of itraconazole. PMID- 9869579 TI - Characterization of dihydrofolate reductase genes from trimethoprim-susceptible and trimethoprim-resistant strains of Enterococcus faecalis. AB - Enterococci are usually susceptible in vitro to trimethoprim; however, high-level resistance (HLR) (MICs, >1,024 microg/ml) has been reported. We studied Enterococcus faecalis DEL, for which the trimethoprim MIC was >1,024 microg/ml. No transfer of resistance was achieved by broth or filter matings. Two different genes that conferred trimethoprim resistance when they were cloned in Escherichia coli (MICs, 128 and >1,024 microg/ml) were studied. One gene that coded for a polypeptide of 165 amino acids (MIC, 128 microg/ml for E. coli) was identical to dfr homologs that we cloned from a trimethoprim-susceptible E. faecalis strain, and it is presumed to be the intrinsic E. faecalis dfr gene (which causes resistance in E. coli when cloned in multiple copies); this gene was designated dfrE. The nucleotide sequence 5' to this dfr gene showed similarity to thymidylate synthetase genes, suggesting that the dfr and thy genes from E. faecalis are located in tandem. The E. faecalis gene that conferred HLR to trimethoprim in E. coli, designated dfrF, codes for a predicted polypeptide of 165 amino acids with 38 to 64% similarity with other dihydrofolate reductases from gram-positive and gram-negative organisms. The nucleotide sequence 5' to dfrF did not show similarity to the thy sequences. A DNA probe for dfrF hybridized under high-stringency conditions only to colony lysates of enterococci for which the trimethoprim MIC was >1,024 microg/ml; there was no hybridization to plasmid DNA from the strain of origin. To confirm that this gene causes trimethoprim resistance in enterococci, we cloned it into the integrative vector pAT113 and electroporated it into RH110 (E. faecalis OG1RF::Tn916DeltaEm) (trimethoprim MIC, 0.5 microg/ml), which resulted in RH110 derivatives for which the trimethoprim MIC was >1, 024 microg/ml. These results indicate that dfrF is an acquired but probably chromosomally located gene which is responsible for in vitro HLR to trimethoprim in E. faecalis. PMID- 9869580 TI - Penetration of oral fluconazole into gynecological tissues. AB - Penetration of fluconazole into female genital tissues was examined. Fluconazole was administered orally at a dose of 150 mg to patients undergoing total abdominal hysterectomy 1 to 151 h prior to surgery. During surgery, blood, uterus, ovary, and oviduct were sampled. Fluconazole concentrations in each tissue were determined by high-performance liquid chromatography. The peak concentrations in serum reached approximately 6.1 microg/ml 1.0 h after a drip infusion was begun. At each time after the infusion, the concentrations in portio vaginalis, cervix uteri, myometrium, endometrium, ovary, and oviduct were higher than those in the serum: the peaks in the tissues ranged from 6.4 to 9.5 microg/g around 1.0 h after the drip infusion was begun. Thus, the levels of penentration of fluconazole into gynecological tissues appeared to be similar to or slightly above those in serum samples. Fluconazole can rapidly penetrate from plasma into the female genital organs, supporting high efficacy of fluconazole against fungal infections in the field of gynecology. PMID- 9869581 TI - Pharmacokinetics of quinupristin-dalfopristin in continuous ambulatory peritoneal dialysis patients. AB - Quinupristin-dalfopristin may be useful for treatment of organisms causing peritoneal dialysis-related peritonitis, including methicillin-resistant coagulase-negative staphylococci, methicillin-resistant Staphylococcus aureus, and vancomycin-resistant enterococci. The pharmacokinetic profiles of single intravenous doses of this combination streptogramin antibiotic of 7.5 mg/kg of body weight were characterized for eight noninfected patients receiving continuous ambulatory peritoneal dialysis. Comparison was made to pharmacokinetic profiles determined for eight healthy volunteers matched by age, sex, and race. Drug was measured in dialysate up to 6 h following the dose. Plasma and dialysate were assayed for parent compounds and metabolites. Mean pharmacokinetic parameters were compared between groups. No statistically significant differences were observed between groups for maximal concentrations in plasma, times to maximal concentration, areas under the curve, distribution volumes, rates of total body clearance, or half-lives in plasma for quinupristin and dalfopristin. No statistically significant differences were observed in maximal concentrations in plasma, times to maximal concentration, areas under the curve, or half-lives for cysteine, the glutathione conjugates of quinupristin, or the pristinamycin IIA metabolite of dalfopristin. The measurements in dialysate of the parent and most metabolites were below the expected MICs. Dialysis clearance was insignificant. Quinupristin-dalfopristin was well tolerated in both groups, causing only mild adverse events that resolved prior to discharge from the study. The disposition of quinupristin, dalfopristin, or their primary metabolites following a single dose was unaltered in patients receiving peritoneal dialysis. Intravenous dosing of this antibiotic combination is unlikely to be adequate for the treatment of peritonitis associated with peritoneal dialysis. PMID- 9869582 TI - Detection of a streptomycin/spectinomycin adenylyltransferase gene (aadA) in Enterococcus faecalis. AB - Genes encoding streptomycin/spectinomycin adenylyltransferases [ANT(3")(9)] have been reported to exist in gram-negative organisms and Staphylococcus aureus. During a study of high-level aminoglycoside resistance in enterococci, we encountered an isolate of Enterococcus faecalis that was streptomycin resistant but did not appear to contain the 6'-adenylyltransferase gene (aadE) when examined by PCR with specific primers. Phosphocellulose paper binding assays indicated the presence of an ANT(3")(9) enzyme. Streptomycin and spectinomycin MICs of 4,000 and 8,000 microg/ml, respectively, were observed for the isolate. PCR primers corresponding to a highly conserved region of the aadA gene were used to amplify a specific 284-bp product. The product hybridized with a digoxigenin labeled PCR product from E. coli C600(pHP45Omega) known to contain the aadA gene. The aadA gene was transferred via filter matings from the E. faecalis donor to E. faecalis JH2-2. PCR primers designed for analysis of integrons were used to amplify a 1-kb product containing the aadA gene, which was cloned into the vector pCRII and transformed into Escherichia coli DH5-alpha competent cells. D Rhodamine dye terminator cycle sequencing was used to determine the gene sequence, which was compared to previously reported sequences of aadA genes. We found the aadA gene in E. faecalis to be identical to the aadA genes reported by Sundstr om et al. for E. coli plasmid R6-5 (L. Sundstrom, P. Radstrom, G. Swedberg, and O. Skold, Mol. Gen. Genet. 213:191-201, 1988), by Fling et al. for the aadA within transposon Tn7 (M. E. Fling, J. Kopf, and C. Richards, Nucleic Acids Res. 13:7095-7106, 1985), and by Hollingshead and Vapnek for E. coli R538-1 (S. Hollingshead and D. Vapnek, Plasmid 13:17-30, 1985). Previous reports of the presence of the aadA gene in enterococci appear to be erroneous and probably describe an aadE gene, since the isolates were reported to be susceptible to spectinomycin. PMID- 9869583 TI - Increase in quinolone resistance in a Haemophilus influenzae strain isolated from a patient with recurrent respiratory infections treated with ofloxacin. AB - The increase in the level of quinolone resistance of Haemophilus influenzae clinical isolates during ofloxacin therapy of a patient with recurrent respiratory infections was investigated. The first isolate (MIC of ciprofloxacin of 2 microg/ml) and the second isolate (MIC of 32 microg/ml) belonged to the same clone, as shown by pulsed-field gel electrophoresis, and the increase in the resistance level was associated with a substitution in Ser-84 to Arg in the ParC protein. These results emphasize the potential risk of development of quinolone resistant H. influenzae during fluoroquinolone therapy in patients with recurrent respiratory infection. PMID- 9869584 TI - Pharmacokinetics in serum and leukocyte exposures of oral azithromycin, 1,500 milligrams, given over a 3- or 5-day period in healthy subjects. AB - The pharmacokinetics in serum and leukocyte (WBC) exposures of 1,500 mg of oral azithromycin administered as 3-day (500 mg/day, days 1 to 3) and 5-day (500 mg on day 1 and 250 mg/day on days 2 to 5) regimens were compared in 12 healthy volunteers. Serum, polymorphonuclear leukocytes, and mononuclear leukocytes were collected over a 12-day period from the start of each regimen. Results of the study indicate that the exposures of serum and both types of WBCs were similar with both regimens. Drug concentrations in day 12 WBCs were well above the MICs for all relevant community-acquired respiratory tract pathogens. Terminal half lives in serum obtained by both regimens were essentially equal at 66 h and consistent with past reports. These results indicate that the standard 1,500-mg dose of oral azithromycin can be administered over either 5 or 3 days. PMID- 9869585 TI - Activity of HMR 3647 compared to those of six compounds against 235 strains of Enterococcus faecalis. AB - Agar dilution was used to test the activities of HMR 3647, erythromycin A, azithromycin, clarithromycin, roxithromycin, clindamycin, and quinupristin dalfopristin against 235 strains of Enterococcus faecalis. HMR 3647 was the most active compound (MICs at which 50 and 90% of the isolates are inhibited [MIC50 and MIC90, respectively] of 0.06 and 4.0 microg/ml, respectively). The MIC50 and MIC90 (with the MIC50 given first and the MIC90 given second; both in micrograms per milliliter) for other compounds were as follows: 4.0 and >32.0 for erythromycin A, 16.0 and >32.0 for azithromycin, 2.0 and >32 for clarithromycin, 32.0 and >32.0 for roxithromycin, 32.0 and >32.0 for clindamycin, and 8.0 and 16.0 for quinupristin-dalfopristin. All compounds were only bacteriostatic. PMID- 9869586 TI - In vitro activities of voriconazole, fluconazole, and itraconazole against 566 clinical isolates of Cryptococcus neoformans from the United States and Africa. AB - We investigated the in vitro activity of voriconazole compared to those of fluconazole and itraconazole against 566 clinical isolates of Cryptococcus neoformans from Africa (164) and the United States (402). Isolates were obtained from cerebrospinal fluid (362), blood (139), and miscellaneous sites (65). Voriconazole (MIC at which 90% of the isolates are inhibited [MIC90], 0.12 to 0.25 microg/ml) was more active than either itraconazole (MIC90, 0.5 microg/ml) or fluconazole (MIC90, 8.0 to 16 microg/ml) against both African and U. S. isolates. Isolates inhibited by >/=16 microg of fluconazole per ml were almost all (99%) inhibited by 500 microg of antibiotic per ml. Cell homogenates generated a novel derivative designated RIP-TAs; in this study, we determined that RIP-TAs is 23-(O-ADP-ribosyl)rifampin. Our results indicated that RIP-TAs is an intermediate in the pathway leading to ribosylated rifampin and that the previously characterized gene encodes a mono(ADP-ribosyl)transferase which, however, shows no sequence similarity to other enzymes of this class. PMID- 9869591 TI - A 5-year surveillance study of 44,691 isolates of Haemophilus influenzae project Beta-Alert 1993-1997. AB - Beta-Alert is a surveillance program developed in 1993 to monitor the percentage of beta-lactamase-producing Haemophilus influenzae isolates obtained from specimens submitted to regional commercial laboratories. The results of this study demonstrate that levels of beta-lactamase producers have remained between 31 and 38% in the United States over the past 5 years. PMID- 9869592 TI - Identification of an efflux pump gene, pmrA, associated with fluoroquinolone resistance in Streptococcus pneumoniae. AB - An open reading frame (ORF) homologous to norA was insertionally inactivated with cat in a fluoroquinolone-resistant pneumococcus with an efflux phenotype; this inactivation caused reversion to drug sensitivity. The ORF product has 24% amino acid sequence identity each to NorA and Bmr, which suggests that it is a major facilitator system pump of the 12-transmembrane-segment class. PMID- 9869593 TI - Metabolic studies on BMS-200475, a new antiviral compound active against hepatitis B virus. AB - BMS-200475 was recently shown to have potent antiviral activity against hepatitis B virus (50% effective concentration = 3.7 nM; 50% cytotoxic concentration = 30 microM). In metabolic studies in both HepG2 and hepatitis B virus-transfected 2.2.15 human hepatoma cell lines, the metabolism was similar, the primary products being the di- and triphosphates. The accumulation of triphosphate was rapid and detectable down to a 5 nM concentration of added drug. When cells were labeled at 25 microM, the intracellular triphosphate concentration attained 30 pmol/10(6) cells ( approximately 30 microM). The intracellular half-life of the triphosphate was about 15 h. Compared with five other nucleoside analogs of medical interest (lamivudine, penciclovir, ganciclovir, acyclovir, and lobucavir), BMS-200475 was most efficiently phosphorylated to the triphosphate in HepG2 cells. PMID- 9869594 TI - In vitro activities of six new fluoroquinolones against Brucella melitensis. AB - We have tested the in vitro activities of eight fluoroquinolones against 160 Brucella melitensis strains. The most active was sitafloxacin (MIC at which 90% of the isolates are inhibited [MIC90], 0.12 microg/ml). In decreasing order, the activities (MIC90s) of the rest of the tested fluoroquinolones were as follows: levofloxacin, 0.5 microg/ml; ciprofloxacin, trovafloxacin, and moxifloxacin, 1 microg/ml; and ofloxacin, grepafloxacin, and gatifloxacin, 2 microg/ml. PMID- 9869596 TI - Clonal analysis of mouse intestinal epithelial progenitors. AB - BACKGROUND & AIMS: Little is known about the cell lineages leading from stem cells to the various terminally differentiated cell types of the intestinal epithelium. In particular, the existence and characterization of intermediate progenitor types remain open issues, which are discussed in this study. METHODS: Chemical mutagenesis was used to genetically mark random intestinal epithelial cells by somatic mutation of the Dlb-1 locus. Intact epithelium was isolated at various times thereafter, and the composition, size, and location of mutant clones were scored. RESULTS & CONCLUSIONS: Analysis of clone dynamics showed short-lived (days) progenitors (C1, M1, and Mix) yielding one or two cell types and long-lived (months) mucous cell progenitors (M0), columnar cell progenitors (C0), and pluripotential stem cells (S) capable of giving rise to all epithelial cell types. Furthermore, study of clonal dispersion, during crypt branching morphogenesis or cell migration, shows that mutant progenitors usually partition into only one of the two daughter crypts and that cells are often widely dispersed in spite of the extensive intercellular junctions in the epithelium. PMID- 9869597 TI - Conjugated bile acid replacement therapy for short-bowel syndrome. AB - BACKGROUND & AIMS: Although fat malabsorption in the short-bowel syndrome is caused in part by decreased bile acid secretion, bile acid replacement therapy is not used because of the belief that ingested bile acids would worsen diarrhea, outweighing the benefits of improved fat absorption. This study compared the effect of a natural conjugated bile acid mixture from ox bile with that of cholylsarcosine, a synthetic conjugated bile acid, on fat absorption and diarrhea in a patient with the short-bowel syndrome. Cholylsarcosine is resistant to bacterial metabolism and has no cathartic activity. METHODS: Metabolic balance studies and a clinical trial were performed in an emaciated patient with the short-bowel syndrome and ileostomy in whom parenteral nutrition could not be used. RESULTS: In balance studies, conjugated bile acid replacement therapy with either preparation caused fat absorption to increase by approximately 40 g/day. Calcium absorption also increased. Neither bile acid product caused a clinically significant increase in ileostomy water output. During a 4-month outpatient trial, while the patient ingested 2 g/meal natural bile acids, her weight increased from 80 to 98 lb, without side effects. CONCLUSIONS: Conjugated bile acid replacement therapy should be part of the armamentarium for the treatment of selected patients with the short-bowel syndrome. PMID- 9869598 TI - Tumor necrosis factor alpha antibody (infliximab) therapy profoundly down regulates the inflammation in Crohn's ileocolitis. AB - BACKGROUND & AIMS: Anti-tumor necrosis factor alpha monoclonal antibody treatment (infliximab) reduces clinical signs and symptoms in patients with Crohn's disease. The effects of infliximab on mucosal histopathologic abnormalities in Crohn's ileocolitis were studied. METHODS: Thirteen patients with steroid refractory Crohn's disease were treated with a single infusion of infliximab (5 20 mg/kg), and 5 were treated with placebo. Ileal and colonic biopsy specimens of all patients were collected before and 4 weeks after therapy. Severity of inflammation was assessed by a histological score. Immunohistochemical stainings with antibodies against HLA-DR, CD68, tumor necrosis factor alpha, intercellular adhesion molecule 1, lymphocyte function-associated antigen, CD4, CD8, and interleukin 4 were performed. RESULTS: Total histological activity score was reduced significantly in both ileitis and colitis after infliximab. This is caused by a virtual disappearance of the neutrophils and a reduction of mononuclear cells. Mucosal architecture returned to normal in 4 patients at 4 weeks. The number of lamina propria mononuclear cells decreased because of a global reduction of CD4(+) and CD8(+) T lymphocytes and CD68(+) monocytes. Aberrant colonic epithelial HLA-DR expression completely disappeared. The percentage of intercellular adhesion molecule 1 and lymphocyte function associated antigen 1-expressing and interleukin 4- and tumor necrosis factor positive lamina propria mononuclear cells sharply decreased. CONCLUSIONS: Infliximab dramatically decreases histological disease activity in Crohn's ileocolitis. Signs of active inflammation nearly disappear accompanied by a profound down-regulation of mucosal inflammatory mediators. PMID- 9869599 TI - Sustained esophageal contraction: a marker of esophageal chest pain identified by intraluminal ultrasonography. AB - BACKGROUND & AIMS: Intraluminal pressure recording systems have not demonstrated predictable esophageal motor correlates of unexplained chest pain. This study used continuous high-frequency intraluminal ultrasonography to characterize esophageal contraction at the time of spontaneous and provoked chest pain. METHODS: Intraluminal pressure, pH, and ultrasound images of the esophagus were recorded for a maximum of 24 hours in 10 subjects with unexplained chest pain. Changes in esophageal muscle thickness were measured as a marker of muscle contraction. Ten additional subjects with suspected esophageal chest pain were studied after edrophonium chloride injection to provoke symptoms. Ten healthy subjects were studied as controls. RESULTS: Eighteen of 24 spontaneous chest pain episodes were preceded by a sustained esophageal contraction (SEC) detected on ultrasonography (mean duration, 68.0 seconds). This motor pattern was not accompanied by changes in intraluminal pressure. Four of 24 asymptomatic control periods were accompanied by SEC, although these contractions were of shorter mean duration (29.0 seconds; P < 0.001). SEC was observed in 5 subjects with a positive chest pain response to edrophonium and in none of the 5 subjects with a negative response. SEC was not detected in normal subjects. CONCLUSIONS: There is a strong temporal correlation between a previously unrecognized esophageal motor event, SEC, and both spontaneous and provoked esophageal chest pain. PMID- 9869600 TI - The kappa agonist fedotozine relieves hypersensitivity to colonic distention in patients with irritable bowel syndrome. AB - BACKGROUND & AIMS: Visceral hypersensitivity plays a major role in the pathophysiology of inflammatory bowel syndrome (IBS). Opioid kappa receptors on afferent nerves may modulate it and may be the target of new IBS treatments. The aim of this study was to evaluate the effects of fedotozine, a potent and selective kappa agonist, on responses to colonic distention and colonic compliance in patients with IBS. METHODS: Fourteen patients with IBS (Rome criteria; 50 +/- 12 years; 6 men and 8 women) were included in a randomized double-blind, crossover trial comparing the effect of an intravenous infusion of 100 mg fedotozine or saline on sensory thresholds elicited by left colon phasic distention (4-mm Hg steps for 5 minutes) up to a sensation of abdominal pain. Colonic compliance was compared by the slope of the pressure-volume curves built on placebo and on fedotozine. RESULTS: In the fedotozine group, thresholds of first perception (28.7 +/- 5.9 mm Hg) and pain (34.7 +/- 5.5 mm Hg) were significantly greater than with placebo (23.3 +/- 4.5 and 29.0 +/- 3.5 mm Hg, respectively; P = 0.0078). Colonic compliance was 9. 20 +/- 3.87 mL. mm Hg-1 with placebo and 8.73 +/- 3.18 mL. mm Hg-1 with fedotozine (not significant). CONCLUSIONS: Fedotozine increases thresholds of perception of colonic distention in patients with IBS without modifying colonic compliance. Fedotozine seems capable of reversing visceral hypersensitivity observed in these patients and could have some beneficial action on their symptoms. PMID- 9869601 TI - Fatty acid chain length determines cholecystokinin secretion and effect on human gastric motility. AB - BACKGROUND & AIMS: Fatty acids induce cholecystokinin (CCK) secretion and modify gastric motility, but the chain length requirements for these effects are not known. Nor is it clear whether the effects of fatty acids on gastric motility in humans are CCK mediated or directly exerted. The aim of this study was to determine the role of fatty acyl chain length in CCK secretion and in influencing gastric motility. METHODS: Fatty acids were infused into the upper gut in healthy volunteers; plasma CCK was determined by radioimmunoassay. Effects of fatty acids on antral contractility were determined by percutaneous ultrasonography; effects on proximal gastric tone were studied during fundal distention. RESULTS: Plasma CCK concentration was consistently and similarly elevated by fatty acids with a chain of 12 carbon atoms or longer, whereas those of 11 or fewer carbon atoms failed to increase plasma CCK. A 12-carbon but not a 10-carbon-long chain fatty acid reduced antral contractile amplitude, an effect that was abolished by loxiglumide (a specific CCK-A receptor antagonist). The 12-carbon fatty acid also reduced proximal gastric tone more than the 10-carbon fatty acid. CONCLUSIONS: A highly specific, chain length-sensitive fatty acid recognition system exists in the proximal gut mediating CCK secretion and gastric motility. An additional, probably CCK-independent, effect of fatty acid also regulates proximal gastric tone. PMID- 9869602 TI - Prevalence of the I1307K APC gene variant in Israeli Jews of differing ethnic origin and risk for colorectal cancer. AB - BACKGROUND & AIMS: Israeli Jews of European birth, i.e., Ashkenazim, have the highest colorectal cancer incidence of any Israeli ethnic group. The I1307K APC gene variant was found in 6.1% of American Jews, 28% of their familial colorectal cancer cases, but not in non-Jews. We assessed the I1307K prevalence in Israeli Jews of differing ethnic origin and risk for colorectal cancer. METHODS: DNA samples from 500 unrelated Jews of European or non-European origin, with or without a personal and/or family history of neoplasia, were examined for the I1307K variant by the allele-specific oligonucleotide (ASO) method. RESULTS: In persons at average risk for colorectal cancer, I1307K was found in 5.0% of 120 European and 1.6% of 188 non-European Jews (P = 0.08). It occurred in 15.4% of 52 Ashkenazi Israelis with familial cancer (P = 0.02) and was not detected in 51 non European Jews at increased cancer risk. Colorectal neoplasia occurred personally or in the families of 13 of 20 Ashkenazi I1307K carriers, 8 of whom also had a personal or family history of noncolonic neoplasia. CONCLUSIONS: The I1307K APC variant may represent a susceptibility gene for colorectal, or other, cancers in Ashkenazi Jews, and partially explains the higher incidence of colorectal cancer in European Israelis. PMID- 9869603 TI - The I1307K polymorphism of the APC gene in colorectal cancer. AB - BACKGROUND & AIMS: Colorectal cancer is one of the most frequent cancers in humans. Recently, a germline missense mutation, I1307K, was identified in the adenomatous polyposis coli (APC) gene that was suggested to increase cancer predisposition in Ashkenazi Jews. However, a second study indicated that the I1307K mutation did not contribute greatly to the risk of colon cancer in Ashkenazi breast-ovarian cancer families, and a role of mismatch repair deficiency was suggested. This study investigated the frequency of the I1307K mutation in several non-Ashkenazi Jewish populations. We also compared the distribution and frequency of APC mutations from colon tumors that were positive and negative for the I1307K mutation. Finally, the association between the presence of mutations in the I1307K region and mismatch repair deficiency was studied. METHODS: We tested for I1307K in 345 patients who were not Ashkenazi Jews using a heteroduplex screen. We also performed an extensive mutational analysis in this region of the APC gene on DNA extracted from 240 Italian, Finnish, and Hawaiian-Japanese colon tumors and determined replication error status. RESULTS: The I1307K mutation was not found among 345 non-Ashkenazis. Somatic mutations occurred at a lower frequency and were more randomly distributed when the I1307K allele was not present. The most common characteristic somatic mutation occurring around codon 1307 in I1307K-positive patients did not occur in tumors negative for the I1307K mutation. An association between mutations in the region around APC codon 1307 and mismatch repair deficiency was not found. CONCLUSIONS: Our findings support the hypothesis that the I1307K mutation is unique to the Ashkenazi Jews, contributes to tumor predisposition in colorectal cancer, and is unrelated to mismatch repair deficiency. PMID- 9869604 TI - Protein kinases C-gamma and -delta are involved in insulin-like growth factor I induced migration of colonic epithelial cells. AB - BACKGROUND & AIMS: The mechanisms by which epithelial cells migrate during the repair of damaged colonic mucosa are poorly understood. This study investigated the insulin-like growth factor I (IGF-I) signaling pathway leading to HT29-D4 human colonic epithelial cell line migration. METHODS: IGF-stimulated cell migration was determined using a wound model in the presence or absence of kinase inhibitors. Activation of protein kinase C (PKC) was determined by immunodetection. RESULTS: IGF-I and insulin induce cell migration without affecting cell proliferation through their cognate receptors. Des(1-3)-IGF-I, a truncated analogue of IGF-I, was more potent than IGF-I, suggesting that IGF binding proteins secreted in the medium modulated IGF-I-induced cell migration. Inhibition of phosphatidylinositol 3-kinase, PKC, and mitogen-activated protein kinases eliminated cell restitution. Long-term exposure of cells to phorbol myristate acetate caused the depletion of PKC-delta and -gamma and prevented also IGF-I-induced cell motility. IGF-I also induced activation of PKC-delta and gamma only. CONCLUSIONS: IGF-I stimulates colonic restitution through the activation of multiple signaling pathways including activation of phosphatidylinositol 3-kinase, PKC-delta and -gamma, and mitogen-activated protein kinases. PMID- 9869605 TI - Gastrin induces heparin-binding epidermal growth factor-like growth factor in rat gastric epithelial cells transfected with gastrin receptor. AB - BACKGROUND & AIMS: Parietal cells express heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF). However, it is unknown whether HB-EGF mediates the trophic action of gastrin. The purpose of this study was to determine whether gastrin modulates the expression of HB-EGF, which mediates the proliferative effects of gastrin on gastric epithelial cells. METHODS: RGM1 cells, a rat gastric epithelial cell line, were transfected with a human gastrin receptor complementary DNA. Gastrin induction of messenger RNAs (mRNAs) for EGF-related polypeptides was assayed by Northern blotting. Processing of cell surface associated proHB-EGF and secretion of HB-EGF were determined by flow cytometry and Western blotting, respectively. Tyrosine phosphorylation of the EGF receptor was assayed by immunoprecipitation and Western blotting with an antiphosphotyrosine antibody. Cell growth was evaluated by [3H]thymidine incorporation. RESULTS: Gastrin induced expression of HB-EGF mRNA, processing of proHB-EGF, release of HB-EGF into the medium, and tyrosine phosphorylation of the EGF receptor. The growth-stimulatory effects of gastrin were partly inhibited by anti-rat HB-EGF serum and completely blocked by AG1478, an EGF receptor-specific tyrphostin. CONCLUSIONS: The findings suggest that HB-EGF at least partially mediates the proliferative effects of gastrin on gastric epithelial cells. PMID- 9869606 TI - Host specificity of Helicobacter pylori strains and host responses in experimentally challenged nonhuman primates. AB - BACKGROUND & AIMS: The specificity of colonization by Helicobacter pylori and complex host-bacterium interactions cannot be readily examined in humans. The aim of this study was to perform such analyses in rhesus monkeys. METHODS: Four animals that had been cured of natural H. pylori colonization were challenged with a mixture of 7 strains of human origin, and bacteria recovered during periodic videogastroscopy were DNA fingerprinted. RESULTS: Three animals carried mixtures of several strains for 4 months, after which strain J166 predominated. In the fourth animal, only strain J238 was isolated from the earliest phase of colonization through 7 months, but strain J166 again became predominant by 10 months after the challenge. Gastritis scores and plasma gastrin and anti-H. pylori immunoglobulin G titers reached levels observed in naturally colonized animals by 4 months after the challenge; however, no plasma immunoglobulin A response was observed up to 10 months. CONCLUSIONS: These results show that (1) natural colonization does not elicit protective immunity against subsequent H. pylori challenge; (2) individuals differ in susceptibility to different H. pylori strains during initial stages of colonization; and (3) certain strains are better suited than others for long-term survival in different hosts. These observations show the complexity of H. pylori-host interactions. PMID- 9869607 TI - Experimental esophagitis induced by acid and pepsin in rabbits mimicking human reflux esophagitis. AB - BACKGROUND & AIMS: The lack of appropriate animal models might explain the paucity of information on the mechanisms of mucosal damage and defense in reflux esophagitis. The aim of this study was to develop a model of esophagitis in rabbits mimicking human reflux esophagitis. METHODS: New Zealand white rabbits underwent surgery for placement of a plastic tube into the cervical esophagus. Acidified pepsin (AP) was intermittently perfused for different periods. Esophageal injury was assessed by macroscopic and microscopic examination, including the cell proliferation immunohistochemical parameter mib1. RESULTS: Rabbit losses (20%) were attributable mostly to postsurgical mortality and tube displacement. Perfusion of AP for 60 min/12 h or 45 min/12 h induced high-grade esophagitis by days 3 and 5, respectively, characterized by diffuse erosion/ulceration, inflammation, bleeding, and reactive epithelial changes. Perfusion of acidified pepsin for 60 min/day, especially at 30 min/12 h, induced low-grade esophagitis characterized by superficial epithelial loss, mild/absent inflammation, and epithelial reactive changes including increased cell proliferation, basal hyperplasia, and papillomatosis, which reached maximal expression by day 7. This perfusion regimen induced mucosal adaptation to damage. CONCLUSIONS: Different and highly reproducible esophageal mucosal lesions mimicking human reflux esophagitis can be induced in rabbits with repetitive acid and pepsin exposure. PMID- 9869608 TI - Functional significance of a newly discovered neuropeptide, orphanin FQ, in rat gastrointestinal motility. AB - BACKGROUND & AIMS: Orphanin FQ (OFQ) is a recently discovered neuropeptide that structurally resembles an opioid peptide. However, the functional role of OFQ in rat gastrointestinal tract remains unknown. METHODS: We investigated the effects of OFQ on contractions of muscle strips obtained from different regions of the gastrointestinal tract. Immunohistochemical studies were performed on rat colonic tissue using OFQ antibody. RESULTS: OFQ (10(-9) to 10(-7) mol/L) caused significant contractions in the rat colon but not in the stomach or small intestine. Tetrodotoxin, veratridine, and long-term serosal application of benzalkonium chloride completely abolished OFQ-induced colonic contractions without affecting myogenic contractions in response to carbachol. OFQ-induced contractions were not affected by naloxone, atropine, phentolamine, propranolol, methysergide, substance P antagonist, vasoactive intestinal polypeptide antagonist, apamin, and NG-nitro-L-arginine methyl ester. OFQ (10(-9) to 10(-7) mol/L) significantly reduced muscle contractions and 3H-acetylcholine release in response to electrical field stimulation in both the stomach and small intestine but not in the colon. OFQ-immunopositive neuronal fibers were found in the colonic myenteric plexus. CONCLUSIONS: These studies indicate that the mechanisms and sites of action of OFQ are region specific. OFQ inhibits cholinergic transmission in the stomach and small intestine, whereas OFQ stimulates colonic contraction possibly by inhibiting an inhibitory neural pathway within the myenteric plexus. PMID- 9869609 TI - Expression of an extracellular calcium-sensing receptor in rat stomach. AB - BACKGROUND & AIMS: Circulating levels of Ca2+ can influence secretory functions and myoelectrical properties of the stomach. A Ca2+-sensing receptor (CaR) has recently been identified in tissues that regulate systemic Ca2+ homeostasis. The aim of this study was to evaluate expression of CaR in the stomach of the rat. METHODS: In forestomach and glandular stomach, reverse-transcription polymerase chain reaction was used to amplify a 380-base pair product, which is 99% homologous with transcripts obtained in parathyroid and kidney. RESULTS: Northern analysis of gastric mucosal polyA+ RNA revealed 7. 5- and 4.1-kilobase transcripts, similar to those obtained in rat parathyroid and kidney. Immunohistochemistry revealed CaR expression in regions of the submucosal plexus and myenteric neurons. In sections of intact tissue, preparations of primary culture surface cells and surgically dissected gastric glands, staining was observed consistently in epithelial cells of the gastric glands and in gastric surface cells. In parietal cells in isolated gastric glands, intracellular levels of Ca2+ responded to conditions that are known to activate CaR. CONCLUSIONS: These are the first reported observations that CaR is expressed in different epithelial cells of mammalian gastric mucosa and its enteric nerve regions. The effects of extracellular Ca2+ on gastric function may be attributable to activation of CaR. PMID- 9869610 TI - The relation of iron status and hemochromatosis gene mutations in patients with chronic hepatitis C. AB - BACKGROUND & AIMS: Elevated hepatic iron concentration may affect the response to antiviral therapy in chronic hepatitis C. This study explored the contribution of genetic hemochromatosis to iron accumulation in chronic hepatitis C. METHODS: HFE mutations (C282Y and H63D) were assessed in 184 patients with chronic hepatitis C virus and 487 controls. Liver biopsy specimens were available in 149 patients. Hepatic iron content was measured in 114 patients by atom-absorption spectrophotometry. RESULTS: The C282Y and H63D allele frequencies were 7.06 and 11.6 in patients and 4.83 and 11.09 in controls, respectively. Eight patients were homozygotes (5 C282Y [2.7%] and 3 H63D [1.6%]), 2 compound heterozygotes (1%), and 49 heterozygotes (14 C282Y [7.6%] and 35 H63D [19%]). Biochemical evidence of iron overload was more common in patients with HFE mutations (28 of 47) than in those without (34 of 102; P = 0.0045). Histological iron grading and hepatic iron content overlapped among patients with or without mutations. A hepatic iron index of >1.9 was observed only in 1 of the 4 C282Y homozygotes and 1 of the 3 H63D homozygotes. CONCLUSIONS: HFE mutations contribute to but do not fully explain hepatic iron accumulation in chronic hepatitis C. Furthermore, C282Y or H63D homozygosity in chronic hepatitis C is not necessarily associated with a high hepatic iron content. PMID- 9869611 TI - Limited humoral immunity in hepatitis C virus infection. AB - BACKGROUND & AIMS: The extremely high rate of chronicity to hepatitis C virus (HVC) infection suggests an inefficient immune response. The humoral immune response to HCV was evaluated in 60 patients with chronic HCV infection and in 12 patients acutely infected with HCV. METHODS: A number of recombinant HCV antigens including the core, envelope 2 (E2), nonstructural (NS) 3, NS4, and NS5 proteins, and NS4a and E2-HVR-1 peptides were used in enzyme-linked immunoassays. RESULTS: Immunoglobulin (Ig) G antibody responses to these viral antigens, except for the HCV core, were highly restricted to the IgG1 isotype. The prevalence of antibodies of the IgG1 isotype specific for the HCV core, E2, E2-HVR1, NS3 (helicase domain), NS4, and NS5 antigens was 97%, 98%, 28%, 88%, 33%, and 68%, respectively. Antibodies of the IgG3 isotype specific for E2, E2-HVR-1, NS3, NS4, and NS5 were detected in a minority of serum samples. The IgG2 and IgG4 isotypes were rarely if ever detected. Furthermore, antibody responses to HCV viral antigens were of relatively low titer and, with the exception of anti-HCV core, were delayed in appearance until the chronic phase of infection. CONCLUSIONS: The IgG1 restriction, low titer, and delayed appearance of antibody responses elicited during HCV infection suggest that the immunogenicity of HCV proteins is limited in the context of natural infection. Inasmuch as recombinant HCV viral antigens perform as relatively normal immunogens in small animals, we suggest that the defective humoral immune responses during HCV infection may be attributable to an "immune avoidance" strategy. PMID- 9869612 TI - Frequent factor II G20210A mutation in idiopathic portal vein thrombosis. AB - BACKGROUND & AIMS: Despite extensive investigations of portal vein thrombosis, no underlying cause is identifiable in up to 30% of patients. A recently described mutation of the prothrombin gene at nucleotide position 20210 is associated with history of venous thrombosis and was assessed in this study. METHODS: We compared the frequency of factor II G20210A and factor V G1691A (factor V Leiden) mutations in 10 patients with idiopathic portal vein thrombosis, 10 patients with nonidiopathic portal vein thrombosis, 60 patients with deep vein thrombosis of the legs, and 42 control subjects. RESULTS: The frequency of factor II G20210A mutation was increased in patients with idiopathic portal vein thrombosis (40.0%; confidence interval, 3.1%-76.9%) compared with controls (4.8%; confidence interval, 0%-11.5%) or patients with nonidiopathic portal vein thrombosis or deep vein thrombosis (P = 0.0001). In contrast, the frequency of the factor V G1691A mutation was similar in subjects with portal vein thrombosis and in controls but was increased in patients with deep vein thrombosis (P = 0.0001). CONCLUSIONS: The factor II G20210A mutation is frequent in patients with idiopathic portal vein thrombosis and should therefore be assessed under this circumstance. PMID- 9869613 TI - UDP-glucuronosyltransferase activity in human liver and colon. AB - BACKGROUND & AIMS: The contribution of glucuronidation toward human drug metabolism is carried out by the Super gene family of UDP glucuronosyltransferases (UGTs). Regulation of the human UGT1A locus is tissue specific, resulting in the unique expression of multiple hepatic and extrahepatic gene products. Studies were undertaken to examine UGT1A expression in human hepatic and colonic tissues. METHODS: UGT1A messenger RNA, protein, catalytic activity, and substrate kinetics were studied in 5 samples of normal hepatic and sigmoid colon tissue using duplex reverse-transcription polymerase chain reaction (RT-PCR), enzymatic and Western blot analysis, and indirect immunofluorescence analysis. RESULTS: Specific patterns of UGT1A gene expression occur in the liver and colon, which were consistent with different banding patterns as detected by Western blot analysis using a UGT1A-specific antibody. However, microsomal UGT activities in colon were up to 96-fold lower for many phenolic substrates, a finding that was not concordant with RT-PCR and Western blot analysis. Interestingly, UGT activity toward tertiary amines and some steroid hormones was equal. CONCLUSIONS: Differences of glucuronidation activity between human liver and colon suggest that UGT1A activity may be regulated as a result of the relative presence of individual isoforms with differing catalytic activities or by tissue-specific modulators after gene expression. PMID- 9869614 TI - Chronic blockade of endothelin receptors in cirrhotic rats: hepatic and hemodynamic effects. AB - BACKGROUND & AIMS: Because controversial roles have been attributed to activation of the hepatic paracrine endothelin (ET) system in cirrhosis, this study assessed whether chronic ET receptor blockade modifies the development of portal hypertension in cirrhotic rats. METHODS: Cirrhotic and control rats received the mixed ET receptor antagonist RO 48-5695 or vehicle daily for 10 weeks. At the end of the treatment period, portal pressure, systemic hemodynamics, standard renal and liver function tests, hepatic concentration of hydroxyproline, and liver messenger RNA (mRNA) expression of procollagen type I were measured. RESULTS: Cirrhotic rats had portal hypertension and hyperdynamic circulation with no differences between animals treated or not treated with the ET-receptor antagonist. However, cirrhotic rats receiving ET-receptor blockade long-term showed a higher hepatic hydroxyproline content and procollagen type I mRNA expression than cirrhotic animals receiving vehicle. CONCLUSIONS: The results indicate that the liver paracrine ET system does not play a major role in the pathogenesis of portal hypertension and support the concept that this system takes part in an autocrine loop that counteracts the development of liver fibrogenesis. PMID- 9869615 TI - Calpain mediates ischemic injury of the liver through modulation of apoptosis and necrosis. AB - BACKGROUND & AIMS: Calpain proteases have been implicated in cell death by necrosis and more recently by apoptosis. Experiments were designed to determine the role of calpain proteases in ischemic rat liver injury by measurement of cytosolic calpain activity after different periods of ischemia-reperfusion and by evaluation of the effects of calpain inhibition on tissue injury and animal survival. METHODS: Calpain activity was measured in the cytosol using Suc-Leu-Leu Val-Try-7 amino-4 methyl coumarin, a specific fluorogenic substrate, and Cbz-Leu Leu-Tyr-CHN2, a specific inhibitor. RESULTS: Calpain activity increased significantly with the duration of ischemia-reperfusion and was inhibited more than 80% by the inhibitor. Calpain inhibition resulted in a significant decrease in transaminase release and tissue necrosis and converted nonsurvival ischemic conditions to survival conditions. When the in situ terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick-end labeling assay for apoptosis was used, 35% +/- 6% of nonparenchymal cells and 16% +/- 3% of hepatocytes stained positively after 60 minutes of ischemia and 6 hours of reperfusion. In contrast, animals pretreated with the calpain inhibitor showed minimal evidence of apoptosis. This was further substantiated by gel electrophoresis assay for DNA fragmentation and by electron-microscopic evaluation. CONCLUSIONS: These data suggest that calpain proteases play a pivotal role in warm ischemia-reperfusion injury of the rat liver through modulation of apoptosis and necrosis. PMID- 9869616 TI - Bile acid feeding induces cholangiocyte proliferation and secretion: evidence for bile acid-regulated ductal secretion. AB - BACKGROUND & AIMS: We have shown that taurocholate (TC) and taurolithocholate (TLC) interact in vitro with normal cholangiocytes, increasing DNA synthesis, secretin receptor (SR) gene expression, and adenosine 3',5'-cyclic monophosphate (cAMP) synthesis. To further extend these in vitro studies, we tested the hypothesis that bile acids (BAs) directly stimulate cholangiocyte proliferation and secretion in vivo. METHODS: After feeding with TC or TLC (1% for 1-4 weeks), we assessed the following in vivo: (1) ductal proliferation by both morphometry and immunohistochemistry for proliferating cell nuclear antigen (PCNA) and measurement of [3H]thymidine incorporation; and (2) the effect of secretin on bile secretion and bicarbonate secretion in vivo. Genetic expression of H3 histone and SR and intracellular cAMP levels were measured in isolated cholangiocytes. RESULTS: After BA feeding, there was an increased number of PCNA positive cholangiocytes and an increased number of ducts compared with control rats. [3H]Thymidine incorporation, absent in control cholangiocytes, was increased in cholangiocytes from BA-fed rats. In BA-fed rats, there was increased SR gene expression (approximately 2.5-fold) and secretin-induced cAMP levels (approximately 3.0-fold) in cholangiocytes, which was associated with de novo secretin-stimulated bile flow and bicarbonate secretion. CONCLUSIONS: These data indicate that elevated BA levels stimulate ductal secretion and cholangiocyte proliferation. PMID- 9869617 TI - Liver transplantation resolves the hyperdynamic circulation in hereditary hemorrhagic telangiectasia with hepatic involvement. AB - BACKGROUND & AIMS: Hepatic involvement in hereditary hemorrhagic telangiectasia is common but often asymptomatic. However, in some cases, the vascular lesions that involve the liver may lead to high-output cardiac failure and pulmonary hypertension that is predominant over hepatobiliary manifestations. Liver transplantation and treatment of these complications are described and discussed in this article. METHODS: Three patients with hereditary hemorrhagic telangiectasia and hepatic involvement received transplants. They had pulmonary hypertension and chronic right-sided heart failure caused by disseminated intrahepatic telangiectasias with shunts between the hepatic artery and hepatic veins or portal vein. Left-to-right intrahepatic shunt output was estimated to range between 51% and 57.5% of cardiac output. RESULTS: Hyperdynamic circulation disappeared after liver transplantation in all patients. Results of computed tomography and right-sided heart catheterization performed 6 months later were normal. Follow-up periods currently are 65, 53, and 29 months, and each patient continues to be asymptomatic. CONCLUSIONS: This report suggests that liver transplantation can be considered as an alternative and successful curative treatment that may prevent the irreversible evolution of cardiopulmonary disease. PMID- 9869618 TI - Molecular medicine and hemochromatosis: at the crossroads. PMID- 9869619 TI - Living and commuting in intestinal crypts. PMID- 9869620 TI - Cancer, crash sites, and the new genetics of neoplasia. PMID- 9869621 TI - The prothrombin G20210A mutation: a new high-prevalence congenital risk factor for thrombosis. PMID- 9869622 TI - The potassium channel and how it works. PMID- 9869623 TI - Dialysis-related amyloidosis: an important cause of gastrointestinal symptoms in patients with end-stage renal disease. PMID- 9869624 TI - Surgeon-related factors and outcome in rectal cancer. PMID- 9869625 TI - The expanding spectrum of clinical uses for botulinum toxin: healing of chronic anal fissures. PMID- 9869627 TI - Posttraumatic stress disorder. PMID- 9869626 TI - High-dose omeprazole in patients with noncardiac chest pain. PMID- 9869628 TI - Gastroenterology fellowship training: a generation lost. PMID- 9869629 TI - Inflammation of the gastric cardia. PMID- 9869630 TI - Polo-like kinases: a team that plays throughout mitosis. PMID- 9869631 TI - Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators. PMID- 9869632 TI - c-myc null cells misregulate cad and gadd45 but not other proposed c-Myc targets. AB - We report here that the expression of virtually all proposed c-Myc target genes is unchanged in cells containing a homozygous null deletion of c-myc. Two noteworthy exceptions are the gene cad, which has reduced log phase expression and serum induction in c-myc null cells, and the growth arrest gene gadd45, which is derepressed by c-myc knockout. Thus, cad and gadd45 are the only proposed targets of c-Myc that may contribute to the dramatic slow growth phenotype of c myc null cells. Our results demonstrate that a loss-of-function approach is critical for the evaluation of potential c-Myc target genes. PMID- 9869633 TI - Transactivation-defective c-MycS retains the ability to regulate proliferation and apoptosis. AB - Transcriptional activation by c-Myc through specific E box elements is thought to be essential for its biological role. However, c-MycS is unable to activate transcription through these elements and yet retains the ability to stimulate proliferation, induce anchorage-independent growth, and induce apoptosis. In addition, c-MycS retains the ability to repress transcription of several specific promoters. Furthermore, c-MycS can rescue the c-myc null phenotype in fibroblasts with homozygous deletion of c-myc. Taken together, our data argue against the paradigm that all of the biological functions of c-Myc are mediated by transcriptional activation of specific target genes through E box elements. PMID- 9869634 TI - Reprogramming of early embryonic blastomeres into endodermal progenitors by a Caenorhabditis elegans GATA factor. AB - The END-1 GATA factor has been implicated in specifying endoderm in Caenorhabditis elegans and is the earliest known zygotic protein expressed in the lineage of E, the clonal endoderm progenitor. We report that ubiquitous end-1 expression during a critical period in embryogenesis causes all non-endodermal lineages to produce endoderm instead of ectoderm and/or mesoderm. END-1 expression bypasses the requirement for maternal SKN-1 and the maternal Wnt signaling pathway in endoderm formation. This suggests that a primary function of these maternal factors is to regulate zygotic end-1 expression, which is then sufficient to initiate the entire program for endoderm development. PMID- 9869635 TI - Molecular interactions between Vestigial and Scalloped promote wing formation in Drosophila. AB - Scalloped (Sd) and Vestigial (Vg) are each needed for Drosophila wing development. We show that Sd is required for Vg function and that altering their relative cellular levels inhibits wing formation. In vitro, Vg binds directly to both Sd and its human homolog, Transcription Enhancer Factor-1. The interaction domains map to a small region of Vg that is essential for Vg-mediated gene activation and to the carboxy-terminal half of Sd. Our observations indicate that Vg and Sd function coordinately to control the expression of genes required for wing development, which implies that Vg is a tissue-specific transcriptional intermediary factor of Sd. PMID- 9869636 TI - Expansion and contraction of ribosomal DNA repeats in Saccharomyces cerevisiae: requirement of replication fork blocking (Fob1) protein and the role of RNA polymerase I. AB - Saccharomyces cerevisiae carries approximately 150 copies of rDNA in tandem repeats. It was found that the absence of an essential subunit of RNA polymerase I (Pol I) in rpa135 deletion mutants triggers a gradual decrease in rDNA repeat number to about one-half the normal level. Reintroduction of the missing RPA135 gene induced a gradual increase in repeat number back to the normal level. Gene FOB1 was shown to be essential for both the decrease and increase of rDNA repeats. FOB1 was shown previously to be required for replication fork blocking (RFB) activity at RFB site in rDNA and for recombination hot-spot (HOT1) activity. Thus, DNA replication fork blockage appears to stimulate recombination and play an essential role in rDNA expansion/contraction and sequence homogenization, and possibly, in the instability of repeated sequences in general. RNA Pol I, on the other hand, appears to control repeat numbers, perhaps by stabilizing rDNA with the normal repeat numbers as a stable nucleolar structure. PMID- 9869637 TI - Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations. AB - To directly determine whether recombinational repair of double-strand breaks (DSBs) can occur between heterologous chromosomes and lead to chromosomal rearrangements in mammalian cells, we employed an ES cell system to analyze recombination between repeats on heterologous chromosomes. We found that recombination is induced at least 1000-fold following the introduction of a DSB in one repeat. Most (98%) recombinants repaired the DSB by gene conversion in which a small amount of sequence information was transferred from the unbroken chromosome onto the broken chromosome. The remaining recombinants transferred a larger amount of information, but still no chromosomal aberrations were apparent. Thus, mammalian cells are capable of searching genome-wide for sequences that are suitable for DSB repair. The lack of crossover events that would have led to translocations supports a model in which recombination is coupled to replication. PMID- 9869638 TI - Maintenance of sister-chromatid cohesion at the centromere by the Drosophila MEI S332 protein. AB - Sister-chromatid cohesion is essential for the faithful segregation of chromosomes during cell division. Recently biochemical analysis with Xenopus extracts suggests that cohesion is established during S phase by a cohesion complex but that other proteins must maintain it in mitosis. The Drosophila melanogaster MEI-S332 protein is present on centromeres in mitosis and meiosis and is essential for cohesion at the centromeres in meiosis II. Here, we analyze the timing of MEI-S332 assembly onto centromeres and the functional domains of the MEI-S332 protein. We find that MEI-S332 is first detectable on chromosomes during prometaphase, and this localization is independent of microtubules. MEI S332 contains two separable functional domains, as mutations within these domains show intragenic complementation. The carboxy-terminal basic region is required for chromosomal localization. The amino-terminal coiled-coil domain may facilitate protein-protein interactions between MEI-S332 and male meiotic proteins. MEI-S332 interacts with itself in the yeast two-hybrid assay and in immunoprecipitates from Drosophila oocyte and embryo extracts. Thus it appears that MEI-S332 assembles into a multimeric protein complex that localizes to centromeric regions during prometaphase and is required for the maintenance of sister-chromatid cohesion until anaphase, rather than its establishment in S phase. PMID- 9869640 TI - The Elongin BC complex interacts with the conserved SOCS-box motif present in members of the SOCS, ras, WD-40 repeat, and ankyrin repeat families. AB - The Elongin BC complex was identified initially as a positive regulator of RNA polymerase II (Pol II) elongation factor Elongin A and subsequently as a component of the multiprotein von Hippel-Lindau (VHL) tumor suppressor complex, in which it participates in both tumor suppression and negative regulation of hypoxia-inducible genes. Elongin B is a ubiquitin-like protein, and Elongin C is a Skp1-like protein that binds to a BC-box motif that is present in both Elongin A and VHL and is distinct from the conserved F-box motif recognized by Skp1. In this report, we demonstrate that the Elongin BC complex also binds to a functional BC box present in the SOCS box, a sequence motif identified recently in the suppressor of cytokine signaling-1 (SOCS-1) protein, as well as in a collection of additional proteins belonging to the SOCS, ras, WD-40 repeat, SPRY domain, and ankyrin repeat families. In addition, we present evidence (1) that the Elongin BC complex is a component of a multiprotein SOCS-1 complex that attenuates Jak/STAT signaling by binding to Jak2 and inhibiting Jak2 kinase, and (2) that by interacting with the SOCS box, the Elongin BC complex can increase expression of the SOCS-1 protein by inhibiting its degradation. These results suggest that Elongin BC is a multifunctional regulatory complex capable of controlling multiple pathways in the cell through interaction with a short degenerate sequence motif found in many different proteins. PMID- 9869639 TI - The RNA cleavage activity of RNA polymerase III is mediated by an essential TFIIS like subunit and is important for transcription termination. AB - Budding yeast RNA polymerase III (Pol III) contains a small, essential subunit, named C11, that is conserved in humans and shows a strong homology to TFIIS. A mutant Pol III, heterocomplemented with Schizosaccharomyces pombe C11, was affected in transcription termination in vivo. A purified form of the enzyme (Pol III Delta), deprived of C11 subunit, initiated properly but ignored pause sites and was defective in termination. Remarkably, Pol III Delta lacked the intrinsic RNA cleavage activity of complete Pol III. In vitro reconstitution experiments demonstrated that Pol III RNA cleavage activity is mediated by C11. Mutagenesis in C11 of two conserved residues, which are critical for the TFIIS-dependent cleavage activity of Pol II, is lethal. Immunoelectron microscopy data suggested that C11 is localized on the mobile thumb-like stalk of the polymerase. We propose that C11 allows the enzyme to switch between an RNA elongation and RNA cleavage mode and that the essential role of the Pol III RNA cleavage activity is to remove the kinetic barriers to the termination process. The integration of TFIIS function into a specific Pol III subunit may stem from the opposite requirements of Pol III and Pol II in terms of transcript length and termination efficiency. PMID- 9869641 TI - Complete inhibition of Cdk/cyclin by one molecule of p21(Cip1). AB - Cell-cycle phase transitions are controlled by cyclin-dependent kinases (Cdks). Key to the regulation of these kinase activities are Cdk inhibitors, proteins that are induced in response to various antiproliferative signals but that can also oscillate during cell-cycle progression, leading to Cdk inactivation. A current dogma is that kinase complexes containing the prototype Cdk inhibitor p21 transit between active and inactive states, in that Cdk complexes associated with one p21 molecule remain active until they associate with additional p21 molecules. However, using a number of different techniques including analytical ultracentrifugation of purified p21/cyclin A/Cdk2 complexes we demonstrate unambiguously that a single p21 molecule is sufficient for kinase inhibition and that p21-saturated complexes contain only one stably bound inhibitor molecule. Even phosphorylated forms of p21 remain efficient inhibitors of Cdk activities. Therefore the level of Cdk inactivation by p21 is determined by the fraction of kinase complexed with the inhibitor and not by the stoichiometry of inhibitor bound to the kinase or the phosphorylation state of the Cdk inhibitor. PMID- 9869642 TI - Lon-mediated proteolysis of the Escherichia coli UmuD mutagenesis protein: in vitro degradation and identification of residues required for proteolysis. AB - Most SOS mutagenesis in Escherichia coli is dependent on the UmuD and UmuC proteins. Perhaps as a consequence, the activity of these proteins is exquisitely regulated. The intracellular level of UmuD and UmuC is normally quite low but increases dramatically in lon- strains, suggesting that both proteins are substrates of the Lon protease. We report here that the highly purified UmuD protein is specifically degraded in vitro by Lon in an ATP-dependent manner. To identify the regions of UmuD necessary for Lon-mediated proteolysis, we performed 'alanine-stretch' mutagenesis on umuD and followed the stability of the mutant protein in vivo. Such an approach allowed us to localize the site(s) within UmuD responsible for Lon-mediated proteolysis. The primary signal is located between residues 15 and 18 (FPLF), with an auxiliary site between residues 26 and 29 (FPSP), of the amino terminus of UmuD. Transfer of the amino terminus of UmuD (residues 1-40) to an otherwise stable protein imparts Lon-mediated proteolysis, thereby indicating that the amino terminus of UmuD is sufficient for Lon recognition and the ensuing degradation of the protein. PMID- 9869643 TI - The Vestigial and Scalloped proteins act together to directly regulate wing specific gene expression in Drosophila. AB - A small number of major regulatory (selector) genes have been identified in animals that control the development of particular organs or complex structures. In Drosophila, the vestigial gene is required for wing formation and is able to induce wing-like outgrowths on other structures. However, the molecular function of the nuclear Vestigial protein, which bears no informative similarities to other proteins, was unknown. Here, we show that Vestigial requires the function of the Scalloped protein, a member of the TEA family of transcriptional regulators, to directly activate the expression of genes involved in wing morphogenesis. Genetic and molecular analyses reveal that Vestigial regulates wing identity by forming a complex with the Scalloped protein that binds sequence specifically to essential sites in wing-specific enhancers. These enhancers also require the direct inputs of signaling pathways, and the response of an enhancer can be switched to another pathway through changes in signal-transducer binding sites. Combinatorial regulation by selector proteins and signal transducers is likely to be a general feature of the tissue-specific control of gene expression during organogenesis. PMID- 9869645 TI - Remnant lipoprotein metabolism: key pathways involving cell-surface heparan sulfate proteoglycans and apolipoprotein E. AB - The plasma clearance of intestinally derived remnant lipoproteins by the liver is a process that likely involves three steps. Our model suggests that the initial rapid clearance by the liver begins with sequestration of the remnants within the space of Disse, where apolipoprotein E secreted by hepatocytes enhances remnant binding and uptake. Heparan sulfate proteoglycans (HSPG), which are also abundant in the space of Disse, mediate this enhanced binding. Next, the remnants undergo further processing in the space of Disse by hepatic and lipoprotein lipases, which may also serve as ligands mediating remnant uptake. The final step, endocytosis by hepatocytes, appears to be mediated, at least in part, by the low density lipoprotein (LDL) receptor and by the LDL receptor-related protein (LRP). Cell-surface HSPG play a critical role in remnant uptake, not only in the important initial sequestration or capture step in the space of Disse, but also as an essential or integral component of the HSPG-LRP pathway. In addition, HSPG appear to function alone as a receptor and display unique handling properties for specific isoforms of apolipoprotein E. PMID- 9869644 TI - Combinatorial signaling codes for the progressive determination of cell fates in the Drosophila embryonic mesoderm. AB - Mesodermal progenitors arise in the Drosophila embryo from discrete clusters of lethal of scute (l'sc)-expressing cells. Using both genetic loss-of-function and targeted ectopic expression approaches, we demonstrate here that individual progenitors are specified by the sequential deployment of unique combinations of intercellular signals. Initially, the intersection between the Wingless (Wg) and Decapentaplegic (Dpp) expression domains demarcate an ectodermal prepattern that is imprinted on the adjacent mesoderm in the form of a L'sc precluster. All mesodermal cells within this precluster are competent to respond to a subsequent instructive signal mediated by two receptor tyrosine kinases (RTKs), the Drosophila epidermal growth factor receptor (DER) and the Heartless (Htl) fibroblast growth factor receptor. By monitoring the expression of the diphosphorylated form of mitogen-associated protein kinase (MAPK), we found that these RTKs are activated in small clusters of cells within the original competence domain. Each cluster represents an equivalence group because all members initially resemble progenitors in their expression of both L'sc and mesodermal identity genes. Thus, localized RTK activity induces the formation of mesodermal equivalence groups. The RTKs remain active in the single progenitor that emerges from each cluster under the subsequent inhibitory influence of the neurogenic genes. Moreover, DER and Htl are differentially involved in the specification of particular progenitors. We conclude that distinct cellular identity codes are generated by the combinatorial activities of Wg, Dpp, EGF, and FGF signals in the progressive determination of embryonic mesodermal cells. PMID- 9869646 TI - The bile acid-inducible baiF gene from Eubacterium sp. strain VPI 12708 encodes a bile acid-coenzyme A hydrolase. AB - The human intestinal Eubacterium sp. strain VPI 12708 has been shown to have a multistep biochemical pathway for bile acid 7alpha-dehydroxylation. A bile acid inducible operon encoding 9 open reading frames has been cloned and sequenced from this organism. Several of the genes in this operon have been shown to catalyze specific reactions in the 7alpha-dehydroxylation pathway. The baiF gene from this operon was cloned, expressed in Escherichia coli, and found to encode a novel bile acid-coenzyme A (CoA) hydrolase. The subunit molecular mass of the purified bile acid-CoA hydrolase was calculated to be 47,466 daltons and the native enzyme had a relative molecular weight of 72,000. The K m and Vmax for cholyl-coenzyme A (CoA) hydrolysis was approximately 175 microm and 374 micromol/min per mg protein, respectively. The enzyme used cholyl-CoA, 3 dehydrocholyl-CoA, and chenodeoxycholyl-CoA as substrates. No hydrolytic activity was detected using acetyl-CoA, isovaleryl-CoA, palmitoyl-CoA, or phenylacetyl-CoA as substrates. Amino acid sequence database searches showed no significant similarity of bile acid-CoA hydrolase to other thioesterases, but significant amino acid sequence identity was found with Escherichia coli carnitine dehydratase. The characteristic thioesterase active site Gly-X-Ser-X-Gly motif was not found in the amino acid sequence of this enzyme. Bile acid-CoA hydrolase from Eubacterium sp. strain VPI 12708 may represent a new family of thioesterases. PMID- 9869648 TI - Regulated turnover of a cell surface-associated pool of newly synthesized apolipoprotein E in HepG2 cells. AB - Newly synthesized apolipoprotein E (apoE) is incompletely secreted and partially degraded by hepatocytes. To evaluate current models concerning apoE's role in remnant lipoprotein clearance by the liver, we performed a detailed and quantitative pulse-chase analysis of apoE secretion, subcellular distribution, and proteolytic degradation by the human hepatoma cell line HepG2. Only about 35% of newly synthesized apoE were found to be secreted to the culture medium. As determined by a protease-protection assay, a substantial amount of newly synthesized apoE was transported to the cell surface, constituting more than half of the cellular apoE under steady-state conditions. A subpopulation representing almost 40% of newly secreted apoE was rapidly rebound to the cell surface, indicating a dynamic equilibrium between cell surface and secreted apoE. These pools of newly synthesized apoE were subject to proteolytic turnover that occurred in lysosomes, presumably after re-endocytosis. We found that the proteolytic turnover of cell surface and secreted apoE was regulated by the availability of apoE ligands, being almost completely suppressed by the presence of lipoprotein-containing human serum or isolated lipoproteins, namely LDL and HDL. The characteristics of regulated degradation of cell surface apoE shed new light on potential physiological functions of this pool of apoE. Our results provide evidence that hepatocytes are capable of forming a large pool of cell surface-associated apoE, thereby supporting the previously proposed secretion recapture model for apoE. PMID- 9869647 TI - HMG-CoA reductase regulation: use of structurally diverse first half-reaction squalene synthetase inhibitors to characterize the site of mevalonate-derived nonsterol regulator production in cultured IM-9 cells. AB - The activity of HMG-CoA reductase (HMGR) is tightly regulated, in part through post-transcriptional mechanisms that are mediated by nonsterol products of mevalonate metabolism. Previous reports have suggested that these mediators are derived from farnesyl pyrophosphate (FPP). Recent studies have implicated FPP hydrolysis products (e.g., farnesol), the squalene synthetase (SQS) reaction products presqualene pyrophosphate (PSQPP) and squalene, or their metabolites. To distinguish among these possible mediators, we evaluated the ability of HMGR and SQS inhibitors to induce compensatory increases in HMGR activity in cultured IM-9 cells. Mevinolin (HMGR inhibitor) produced predicted increases in HMGR activity that were related to the degree of cholesterolgenesis inhibition (e.g., 4-fold, 9 fold, and 17-fold increases relative to 50%, 76%, and 90% inhibition, respectively). By contrast, a variety of structurally distinct reversible, competitive, first half-reaction SQS inhibitors all reduced cholesterolgenesis by up to 90% with no appreciable increases in HMGR activity. These observations strongly suggest that nonsterol-mediated post-transcriptional mechanisms regulating HMGR activity remain intact after SQS first half-reaction inhibition, indicating that nonsterol regulator production is independent of SQS action and ruling out PSQPP, squalene and their metabolites as possible mediators. Unexpectedly, the SQS mechanism-based irreversible inactivator, zaragozic acid A (ZGA) exhibited the greatest degree of HMGR modulation, producing 5-fold, 11 fold, and 40-fold increases in HMGR activity at concentrations that produced 25%, 50%, and 75% cholesterolgenesis inhibition, respectively. The markedly greater magnitude of HMGR stimulation by ZGA versus mevinolin at similar levels of cholesterolgenesis inhibition suggests that ZGA may directly interfere with the production or action of the nonsterol regulator. PMID- 9869649 TI - A common Hpa I RFLP of apolipoprotein C-I increases gene transcription and exhibits an ethnically distinct pattern of linkage disequilibrium with the alleles of apolipoprotein E. AB - Apolipoprotein (apo) C-I is a constituent of triglyceride-rich lipoproteins (TGRL) that interferes with their hepatic clearance. Functional polymorphism in the apoC-I gene has not been established. We determined that an Hpa I site variably present at -317 relative to the apoC-I gene is produced by a 4-bp CGTT insertion. The apoC-I Hpa I alleles showed an ethnically distinct pattern of linkage disequilibrium with the alleles of the adjacent apoE gene. The frequency of apoC-I Hpa I-positive (H2) with apoE varepsilon2 was 0. 98, without significant ethnic difference. In contrast, the frequency of H2 with apoE epsilon4 was 0.85 in European-Americans but only 0.55 in African-Americans (P < 0.001). The frequency of H2 with apoE epsilon3 was 0.02 in European-Americans and 0.08 in African-Americans (P < 0.001). African-American apoE epsilon3/epsilon3 carriers of apoC-I H2 had 19% lower fasting triglyceride levels than H1 homozygotes (P = 0.03) along with 18% higher HDL-cholesterol levels (P = 0.02). ApoB levels were 21% lower (P = 0.002). H2-allelic reporter-gene constructions showed 50% higher expression in transient transfection studies. We localized the source of this difference in expression to the CGTT insertion itself. Deletion studies of the H1 allele showed a negative transcriptional effect of the polymorphic region. An H1 oligodeoxynucleotide showed specific binding of a hepatoma-cell nuclear protein not evident with an H2 oligodeoxynucleotide. The H2 sequence may decrease the binding of a negatively acting transcription factor, leading to overexpression of apoC-I. This may produce a functional effect on lipoprotein levels but confirmation is needed in other populations. PMID- 9869650 TI - Effects of natural mutations in lecithin:cholesterol acyltransferase on the enzyme structure and activity. AB - A molecular model was built for human lecithin:cholesterol acyltransferase (LCAT) based upon the structural homology between this enzyme and lipases (Peelman et al. 1998. Prot. Sci. 7: 585-597). We proposed that LCAT belongs to the alpha/beta hydrolase fold family, and that the central domain of LCAT consists of a mixed seven-stranded beta-pleated sheet with four alpha-helices and loops linking the beta-strands. The catalytic triad of LCAT was identified as Asp345 and His377, as well as Ser181. This model is used here for the interpretation of the structural defects linked to the point mutations identified in LCAT, which cause either familial LCAT deficiency (FLD) or fish-eye disease (FED). We show that these mutations occur in separate domains of the 3D structure of the enzyme. Most mutations causing familial LCAT deficiency are either clustered in the vicinity of the catalytic triad or affect conserved structural elements in LCAT. Most mutations causing fish-eye disease are localized on the outer hydrophilic surface of the amphipathic helical segments. These mutations affect only minimally the overall structure of the enzyme, but are likely to impair the interaction of the enzyme with its co-factor and/or substrate. PMID- 9869651 TI - 3-Hydroxy-3-methylglutaryl coenzyme A lyase: targeting and processing in peroxisomes and mitochondria. AB - 3-Hydroxy-3-methylglutaryl coenzyme A lyase (HL, E.C. 4.1.3.4) has a unique dual localization in both mitochondria and peroxisomes. Mitochondrial HL ( approximately 31.0 kDa) catalyzes the last step of ketogenesis; the function of peroxisomal HL ( approximately 33.5 kDa) is unknown. On density gradient fractionation, normal human lymphoblasts contain both peroxisomal and mitochondrial HL whereas in lymphoblasts from a patient with Zellweger syndrome, in which functional peroxisomes are absent, only the mitochondrial HL isoform was present. To study the kinetics of the dual targeting of HL, we performed pulse chase experiments in normal and Zellweger cells. Pulse-chase studies revealed a biphasic curve for processing of the HL precursor. The first phase, with a calculated half-life of approximately 3 h in both normal and Zellweger fibroblasts and lymphoblasts and in HepG2 cells, presumably reflects mitochondrial import and processing of the precursor; the second (t1/2, 12-19 h) is present only in normal cells and presumably represents the half-life of peroxisomal HL. The half-life of mature mitochondrial HL was 14 to 19 h in both normal and Zellweger cells. Studies of the HMG-CoA lyase precursor in isolated rat mitochondria showed a rate of processing approximately 2.6-fold lower than that of the ornithine transcarbamylase precursor. PMID- 9869652 TI - Generation of monoclonal antibodies specific for mouse apolipoprotein B-100 in apolipoprotein B-48-only mice. AB - Over the past 10 years, many laboratories have investigated lipid metabolism and atherogenesis with a variety of transgenic and gene knockout mouse models. Although many of these studies have yielded valuable insights, some have been hampered by a paucity of useful antibodies against mouse proteins. For example, many laboratories have analyzed genetic and dietary interventions affecting lipoprotein metabolism without useful antibodies against mouse apolipoprotein (apo) B. In this study, we sought to develop highly specific monoclonal antibodies against mouse apoB-100. To achieve this goal, gene-targeted mice that synthesize exclusively apoB-48 (apoB-48-only mice) were immunized with mouse apoB 100. The immune response against apoB-100 was robust, as judged by high titers of antibodies against mouse apoB-100. After fusing the splenic lymphocytes of the apoB-48-only mice with a myeloma cell line, we identified and cloned hybridomas that produced mouse apoB-100-specific monoclonal antibodies. Those antibodies were useful for developing sensitive and specific immunoassays for mouse apoB 100. This study illustrates the feasibility and utility of using gene-targeted mice to develop monoclonal antibodies against mouse proteins. PMID- 9869653 TI - Apolipoprotein A-I promotes cholesterol release and apolipoprotein E recruitment from THP-1 macrophage-like foam cells. AB - Apolipoprotein E (apoE) is synthesized and secreted by arterial macrophages while apolipoprotein A-I (apoA-I) is present in surrounding interstitial fluids. Both apolipoproteins play important roles in macrophage cholesterol homeostasis by forming lipid complexes (nascent-HDL) with cellular phospholipids (PL) and cholesterol (UC) thereby promoting cholesterol efflux. In this study, we evaluated the relative contributions of apoA-I and endogenously produced apoE in mediating the recruitment of cellular cholesterol. THP-1 human monocytes were differentiated (300 nm phorbol dibutyrate) into macrophages and macrophage-foam cells were generated by cholesterol loading with acetylated LDL (50 microg protein/ml). ApoA-I (10 microg/ml) depleted macrophage-foam cell cholesteryl esters by 50% in 24 h. This reduction was accompanied by a significant increase in the UC/PL mole ratio of nascent HDL (UC/PL = 0.80 +/- 0.15) in the medium compared to complexes isolated from macrophages (UC/PL = 0.59 +/- 0.08). Significantly more (70%) nascent-HDL were formed in incubations of apoA-I with macrophage-foam cells than with macrophages. Medium apoE accumulation paralleled the assembly of apoA-I containing nascent HDL where 2- and 4-fold increases were observed with macrophages and macrophage-foam cells, respectively, compared to incubations in the absence of apoA-I. Despite the increase in medium apoE accumulation, a majority (85%) of particles (11, 9, and 7.4 nm in diameter) from macrophages and macrophage-foam cells possessed apoA-I without apoE. ApoA-I plus apoE particles (13-16 nm) were also formed along with a small quantity of apoE only particles (19-20 nm). The predominance of apoA-I only particles indicates, however, that the assembly of apoA-I-containing nascent-HDL represents a major metabolic pathway of cellular cholesterol recruitment compared to the endogenous production of apoE. PMID- 9869654 TI - Lipid binding-induced conformational changes in the N-terminal domain of human apolipoprotein E. AB - The N-terminal domain of human apolipoprotein E3 (apoE3) adopts an elongated, globular four helix bundle conformation in the lipid-free state. Upon lipid binding, the protein is thought to undergo a significant conformational change that is essential for manifestation of its low density lipoprotein receptor recognition properties. We have used fluorescence resonance energy transfer (FRET) to characterize helix repositioning which accompanies lipid interaction of this protein. ApoE3(1-183) possesses a single cysteine at position 112 and four tryptophan residues (positions 20, 26, 34, and 39). Modification of Cys112 with the chromophore, N-iodoacetyl-N'-(5-sulfo-1-naphthyl)etheylenediamine (AEDANS) was specific and did not alter the secondary structure content of the protein. The efficiency of energy transfer from donor Trp residues to the AEDANS moiety was 49% in buffer, consistent with close proximity of the chromophores. Guanidine HCl titration experiments induced characteristic changes in the efficiency of energy transfer, indicating that FRET data faithfully reports on the conformational status of the protein. Interaction of AEDANS-apoE3(1-183) with dimyristoylphosphatidylcholine to form disk particles, or with detergent micelles, resulted in large decreases in the efficiency of energy transfer. Distance calculations based on the FRET measurements revealed that lipid binding increases the average distance between the four Trp donors and the AEDANS acceptor from 23 A to 44 A. The results obtained demonstrate the utility of FRET to investigate conformational adaptations of exchangeable apolipoproteins and are consistent with the hypothesis that, upon lipid binding, apoE3(1-183) undergoes conformational opening, repositioning helix 1 and 3 to adopt a receptor-active conformation. PMID- 9869655 TI - Differential effects of 17alpha-ethinylestradiol on the neutral and acidic pathways of bile salt synthesis in the rat. AB - Effects of 17alpha-ethinylestradiol (EE) on the neutral and acidic biosynthetic pathways of bile salt (BS) synthesis were evaluated in rats with an intact enterohepatic circulation and in rats with long-term bile diversion to induce BS synthesis. For this purpose, bile salt pool composition, synthesis of individual BS in vivo, hepatic activities, and expression levels of cholesterol 7alpha hydroxylase (CYP7A), and sterol 27-hydroxylase (CYP27), as well as of other enzymes involved in BS synthesis, were analyzed in rats treated with EE (5 mg/kg, 3 days) or its vehicle. BS pool size was decreased by 27% but total BS synthesis was not affected by EE in intact rats. Synthesis of cholate was reduced by 68% in EE-treated rats, while that of chenodeoxycholate was increased by 60%. The recently identified Delta22-isomer of beta-muricholate contributed for 5.4% and 18.3 % (P < 0.01) to the pool in control and EE-treated rats, respectively, but could not be detected in bile after exhaustion of the pool. A clear reduction of BS synthesis was found in bile-diverted rats treated with EE, yet biliary BS composition was only minimally affected. Activity of CYP7A was decreased by EE in both intact and bile-diverted rats, whereas the activity of the CYP27 was not affected. Hepatic mRNA levels of CYP7A were significantly reduced by EE in bile diverted rats only; CYP27 mRNA levels were not affected by EE. In addition, mRNA levels of sterol 12alpha-hydroxylase and lithocholate 6beta-hydroxylase were increased by bile diversion and suppressed by EE. This study shows that 17alpha ethinylestradiol (EE)-induced intrahepatic cholestasis in rats is associated with selective inhibition of the neutral pathway of bile salt (BS) synthesis. Simultaneous impairment of other enzymes in the BS biosynthetic pathways may contribute to overall effects of EE on BS synthesis. PMID- 9869656 TI - Chinese hamster ovary cells overexpressing the oxysterol binding protein (OSBP) display enhanced synthesis of sphingomyelin in response to 25-hydroxycholesterol. AB - 25-Hydroxycholesterol negatively regulates cholesterol synthesis and activates cholesterol esterification in a variety of cultured cells. Concurrent with these effects, 25-hydroxycholesterol also stimulates the synthesis of sphingomyelin in Chinese hamster ovary (CHO)-K1 cells. The role of oxysterol binding protein (OSBP), a high affinity receptor for 25-hydroxycholesterol, in activation of SM synthesis was assessed by overexpression in CHO-K1 cells. When compared to mock transfected controls, three CHO-K1 clones overexpressing OSBP by 10- to 15-fold displayed a 2- to 3-fold enhancement of [3H]serine incorporation into sphingomyelin when treated with 25-hydroxycholesterol. Closer examination of one of these clones (CHO-OSBP cells) revealed a >8.5-fold stimulation of sphingomyelin synthesis after a 6-h treatment with 25-hydroxycholesterol compared to 3.5-fold in controls, slightly higher basal levels of sphingomyelin synthesis, and a more rapid response to 25-hydroxycholesterol. [3H]serine incorporation into phosphatidylserine, phosphatidylethanolamine, ceramide, or glucosylceramide was affected by <15%. Synthesis of sphingomyelin from exogenous [3H]sphinganine labeled ceramide was enhanced in overexpressing cells treated with 25 hydroxycholesterol. However, in vitro activities of sphinganine N acyltransferase, sphingomyelin synthase, and serine palmitoyltransferase were not affected by OSBP overexpression or 25-hydroxycholesterol. Overexpression of OSBP or 25-hydroxycholesterol did not significantly affect the ceramide content of Golgi-enriched fractions from control or overexpressing cells. However, diglyceride mass was reduced in Golgi-enriched fractions from overexpressing cells and by treatment with 25-hydroxycholesterol. Results from overexpressing cells show that OSBP potentiates the stimulatory effects of 25-hydroxycholesterol on sphingomyelin synthesis. 25-Hydroxycholesterol promotes translocation of OSBP to the Golgi apparatus where it appears to stimulate conversion of ceramide to sphingomyelin. PMID- 9869657 TI - Chemical synthesis of D-ribo-phytosphingosine-1-phosphate, a potential modulator of cellular processes. AB - d-erythro -Sphingosine-1-phosphate (2), an intermediate in sphingosine metabolism, shows a diversity of biological activities. Comparable roles might be anticipated for d-ribo -phytosphingosine-1-phosphate (1). We describe an efficient three-step chemical synthesis of 1 from d-ribo -phytosphingosine. Our approach is based on standard phosphoramidite methodology and on the finding of Boumendjel and Miller ( J. Lipid Res. 1994. 35: 2305-2311) that sphingosine can be monophosphorylated at the 1-hydroxyl without protection of the 3-hydroxyl. However, we were unable to duplicate their reported synthesis of 2 without important modifications in reagents and reaction conditions. Under the reported conditions for preparing 2, we obtained a cyclic carbamate (14), which we have isolated and identified. The structures of 1 and the cyclic carbamate 14 were elucidated by a combination of mass spectrometry and 1D and 2D nuclear magnetic resonance spectroscopy. PMID- 9869658 TI - Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry. AB - A mass spectrometric method for the quantification of free cholesterol in cells and subcellular membranes is presented. The method is based on a simple one-step chemical derivatization of cholesterol to cholesterol-3-sulfate by a sulfur trioxide-pyridine complex. Quantification is performed by nano-electrospray ionization tandem mass spectrometry (nanoESI-MS/MS) using a stable isotope labeled internal standard. The determination of free cholesterol is demonstrated in about 250 cells of a Chinese hamster ovary (CHO) cell line. With this method a molar ratio of free cholesterol to total phospholipids of 0.34 mol/mol in CHO cells was determined. In a subcellular membrane fraction enriched in Golgi membranes, a molar ratio of free cholesterol to total phospholipids of 0.57 mol/mol was determined. The method should be of value for quantification of other sterols as demonstrated for ergosterol and stigmasterol. PMID- 9869659 TI - Paraoxonase protection of LDL against peroxidation is independent of its esterase activity towards paraoxon and is unaffected by the Q-->R genetic polymorphism. AB - High density lipoprotein (HDL)-associated paraoxonase (PON) seems to play a major role in the protection of low density lipoprotein (LDL) against peroxidation by HDL, and the partly purified enzyme exerts a dose-dependent protective effect. A common polymorphism of the human gene (192 Q-->R) modulates paraoxonase activity but purified enzyme from either genotype is equally effective against LDL peroxidation. The inhibition of Cu2+-induced LDL peroxidation by HDL was monitored by lipid peroxide assay and change in LDL electrophoretic mobility. We show that HDL from type 2 diabetic patients with the QQ or RR genotype (n = 12 for each) reduce, to the same extent, both peroxide production (by 60.6 +/- 20.0 and 63.9 +/- 23.5%) and relative change in mobility (61.3 +/- 21.8 and 61.4 +/- 26.5%) despite a 6-fold difference in paraoxonase activity (47.4 +/- 4.4 vs. 299.7 +/- 23.7 U/l, P < 0.0001). Protection was, however, related to paraoxonase activity, but with a different efficiency in each group corresponding to a better protection per unit of enzyme in the QQ genotype group. Inactivation of PON activity by heating (56 degreesC, 10 min) or by EDTA was totally without effect on protection, which remained correlated with the paraoxonase activity measured prior to inactivation. In summary, these results suggest that the protein bearing both paraoxonase and arylesterase activities also possesses a third thermostable property, closely associated with paraoxon hydrolysis activity and unaffected by PON genetic variability. PMID- 9869660 TI - Structural analyses of polymorphic transitions of sn-1, 3-distearoyl-2 oleoylglycerol (SOS) and sn-1, 3-dioleoyl-2-stearoylglycerol (OSO): assessment on steric hindrance of unsaturated and saturated acyl chain interactions. AB - Polymorphic transformations in two saturated-unsaturated mixed acid triacylglycerols, SOS (sn -1,3-distearoyl-2-oleoylglycerol) and OSO (sn -1,3 dioleoyl-2-stearoylglycerol), have been studied by FT-IR spectroscopy using deuterated specimens in which stearoyl chains are fully deuterated. A reversible phase transition between sub alpha and alpha and a series of irreversible transitions (alpha-->gamma-->beta'-->beta (beta2, beta1) for SOS and alpha- >beta'-->beta for OSO) were studied with an emphasis on the conformational ordering process of stearoyl and oleoyl chains. The alpha-->sub alpha reversible transition was due to the orientational change of stearoyl chains in the lateral directions from the hexagonal subcell to a perpendicularly packed one. As the first stage of the series of irreversible transitions from alpha to beta, the conformational ordering of saturated chains took place in the alpha-->gamma transition of SOS and in the alpha-->beta' transition of OSO; one stearoyl chain in SOS and OSO takes the all-trans conformation and the second stearoyl chain in SOS takes the bent conformation like those observed in the most stable beta-type. As the final stage, the ordering of unsaturated chains occurred in the beta'- >beta transition both for SOS and OSO. A conversion in the layered structure from bilayer to trilayer was also accompanied by the conformational ordering in the alpha-->gamma transition of SOS and in the beta'-->beta transition of OSO. PMID- 9869661 TI - In vivo involvement of cytochrome P450 4A family in the oxidative metabolism of the lipid peroxidation product trans-4-hydroxy-2-nonenal, using PPARalpha deficient mice. AB - Trans-4-hydroxy-2-nonenal (HNE) is a potent cytotoxic and genotoxic compound originating from the peroxidation of n-6 polyunsaturated fatty acids. Its metabolism has been previously studied in the rat (Alary et al. 1995. Chem. Res. Toxicol., 8: 35-39). In addition to major urinary mercapturic derivatives, some polar urinary metabolites were isolated and could correspond to hydroxylated compounds. 4-Hydroxynonenoic acid (HNA), resulting from the oxidation of the HNE carbonyl group, is a medium chain fatty acid and its omega-hydroxylation might be hypothesized. Therefore, the involvement of the CYP 4A family isoenzymes in the metabolism of [3H]HNE has been investigated in vivo using inducer treatments (fibrates) in wild-type or in peroxisome proliferator-activated receptor alpha (PPARalpha)-deficient mice. In wild-type mice, but not in PPARalpha (-/-) mice, fibrate treatments resulted in an increase of two urinary metabolites characterized, after HPLC purifications and mass spectrometry analyses, as the omega-hydroxylated metabolite of HNA, i.e., 4,9-dihydroxy-2-nonenoic acid, and its oxidized form, 4-hydroxy-2-nonene-1,9-dicarboxylic acid. The formation of the latter is correlated accurately to laurate hydroxylase activity studied concurrently in microsomes prepared from the liver of these animals. Basal levels of these two metabolites were measured in urine of normal and PPARalpha-deficient mice. These results are in accord with an implication of the P450 4A family in the extended oxidative metabolism of 4-HNE. PMID- 9869662 TI - A simple, nonenzymatic method for desialylating polysialylated ganglio-N-tetraose series gangliosides to produce GM1. AB - Dowex-50W-H+ was used to catalyze the highly selective desialylation of polysialylated ganglio-N-tetraose series gangliosides to yield primarily GM1. High performance thin-layer chromatographic analysis of recovered lipid indicated that 60-70% of the recovered ganglioside was GM1. Identification of the major product as GM1 was confirmed by proton NMR spectra and lack of sialic acid release by Vibrio cholerae sialidase. PMID- 9869663 TI - Rapid separation and identification of phosphatidylethanolamine molecular species. AB - A novel high performance liquid chromatography method is presented for the separation and identification of intact molecular species of phosphatidylethanolamine (PE). After isocratic separation, detection of species can be achieved by measurement of UV absorbance as well as by the quantitative method of light scattering detection. A mathematical relationship exists between i) the relative retention time of a PE molecular species and ii) the number of carbon atoms and double bonds in the aliphatic groups of the species. This relationship can aid in the identification of the species. Furthermore, the absence of non-volatile components in the solvent allows the use of electrospray mass spectrometry to identify the eluting components and to establish the position of the individual radyl groups at the glycerol backbone. Using this method, samples of bovine heart PE (rich in plasmalogens) and rat liver PE (rich in diacyl species) have been analyzed. PMID- 9869664 TI - Analytical capillary isotachophoresis of total plasma lipoproteins: a new tool to identify atherogenic low density lipoproteins. AB - Plasma low density lipoproteins from 20 patients were separated by capillary isotachophoresis (ITP). In each patient the apparent diameter of the predominant LDL peak on whole plasma was also determined by nondenaturing gradient gel electrophoresis. Furthermore the concentration of the more electronegatively charged in vivo oxidized LDL- was accomplished using anion exchange high pressure liquid chromatography. By analytical capillary ITP of whole plasma lipoproteins, prestained with a lipophilic dye, LDL were separated into four subfractions. Usually, the predominant subfraction was the slow migrating LDL4, followed by LDL3, and then by the faster LDL2 and LDL1. Slow migrating LDL4 correlated negatively with plasma triglycerides and LDL- and positively with plasma high density lipoprotein (HDL) cholesterol and with the LDL diameter, while the faster LDL1 showed an inverse behavior. The LDL1 + LDL2 to LDL3 + LDL4 ratio showed a strong positive correlation with LDL- concentration (r = 0.87; P < 0.001) and a highly significant inverse correlation with the LDL particle diameter (r = -0.74; P < 0.001). At least three highly atherogenic LDL that could be found in human plasma, namely oxidized, glycated and small-dense, are characterized by a greater electric charge. The LDL profile from capillary ITP and the relative prevalence of faster or slower migrating LDL fractions could indicate the presence of more atherogenic LDL. PMID- 9869665 TI - Cloning and mutational analysis of human malonyl-coenzyme A decarboxylase. AB - Malonyl coenzyme A (CoA) decarboxylase (E.C.4. 1.1.9) catalyzes the conversion of malonyl CoA to acetyl CoA. The metabolic role of malonyl CoA decarboxylase has not been fully defined, but deficiency of the enzyme has been associated with mild mental retardation, seizures, hypotonia, cardiomyopathy, vomiting, hypoglycemia, metabolic acidosis, and malonic aciduria. Here we report the isolation and sequencing of the human gene encoding malonyl CoA decarboxylase, and the identification of a mutation causing malonyl CoA decarboxylase deficiency. Human malonyl CoA decarboxylase cDNA sequences were identified by homology to the goose gene, and the intron/exon boundaries were determined by direct sequencing of a PAC clone containing the entire human gene. The 1479 basepair human cDNA is 70 percent identical to the goose sequence, and the intron/exon boundaries are completely conserved between the two species. The genetic mutation underlying malonyl CoA decarboxylase deficiency was determined in a patient with clinical features of this defect, malonic aciduria, and markedly reduced malonyl CoA decarboxylase activity. PMID- 9869666 TI - Inadequate management of blood pressure in a hypertensive population. AB - BACKGROUND: Many patients with hypertension have inadequate control of their blood pressure. Improving the treatment of hypertension requires an understanding of the ways in which physicians manage this condition and a means of assessing the efficacy of this care. METHODS: We examined the care of 800 hypertensive men at five Department of Veterans Affairs sites in New England over a two-year period. Their mean (+/-SD) age was 65.5+/-9.1 years, and the average duration of hypertension was 12.6+/-5.3 years. We used recursive partitioning to assess the probability that antihypertensive therapy would be increased at a given clinic visit using several variables. We then used these predictions to define the intensity of treatment for each patient during the study period, and we examined the associations between the intensity of treatment and the degree of control of blood pressure. RESULTS: Approximately 40 percent of the patients had a blood pressure of > or =160/90 mm Hg despite an average of more than six hypertension related visits per year. Increases in therapy occurred during 6.7 percent of visits. Characteristics associated with an increase in antihypertensive therapy included increased levels of both systolic and diastolic blood pressure at that visit (but not previous visits), a previous change in therapy, the presence of coronary artery disease, and a scheduled visit. Patients who had more intensive therapy had significantly (P<0.01) better control of blood pressure. During the two-year period, systolic blood pressure declined by 6.3 mm Hg among patients with the most intensive treatment, but increased by 4.8 mm Hg among the patients with the least intensive treatment. CONCLUSIONS: In a selected population of older men, blood pressure was poorly controlled in many. Those who received more intensive medical therapy had better control. Many physicians are not aggressive enough in their approach to hypertension. PMID- 9869667 TI - Value of electron-beam computed tomography for the noninvasive detection of high grade coronary-artery stenoses and occlusions. AB - BACKGROUND: Reliable noninvasive assessment of coronary-artery stenoses and occlusions would constitute an advantage in the care of patients with known or suspected coronary artery disease. We investigated the accuracy of contrast enhanced electron-beam computed tomography (CT) for the detection of high-grade coronary-artery stenoses and occlusions. METHODS: Electron-beam CT was performed in 125 patients. After intravenous injection of a contrast agent, 40 cross sectional images of the heart were acquired during inspiration, triggered by the electrocardiogram in diastole. Three-dimensional reconstructions of the heart and coronary arteries were rendered to facilitate evaluation of the images. The proximal and middle segments of the major coronary arteries were evaluated for the presence or absence of high-grade stenoses and occlusions. The results were compared with those of invasive coronary angiography in a blinded fashion. RESULTS: Because of technical problems that impaired the quality of the images, 124 (25 percent) of the 500 coronary arteries studied (left main, left anterior descending, left circumflex, and right coronary) in a total of 125 patients were excluded from evaluation. No vessels could be evaluated in 19 patients (15 percent), and another 28 patients (22 percent) had one, two, or three vessels that could not be evaluated. In the remaining coronary arteries with adequate image quality, electron-beam CT permitted visualization of 69 of 75 high-grade stenoses and occlusions (sensitivity, 92 percent), whereas in 282 of 301 arteries, the absence of high-grade stenoses and occlusions was correctly detected (specificity, 94 percent). CONCLUSIONS: When image quality is adequate, electron-beam CT may be useful to detect or rule out high-grade coronary-artery stenoses and occlusions. PMID- 9869668 TI - Effect of HMG-CoA reductase inhibitors on coronary artery disease as assessed by electron-beam computed tomography. AB - BACKGROUND: Angiographic studies of the regression of coronary artery disease are invasive and costly, and they permit only limited assessment of changes in the extent of atherosclerotic disease. Electron-beam computed tomography (CT) is noninvasive and inexpensive. The entire coronary-artery tree can be studied during a single imaging session, and the volume of coronary calcification as quantified with this technique correlates closely with the total burden of atherosclerotic plaque. METHODS: We conducted a retrospective study of 149 patients (61 percent men and 39 percent women; age range, 32 to 75 years) with no history of coronary artery disease who were referred by their primary care physicians for screening electron-beam CT. All patients underwent base-line scanning and follow-up assessment after a minimum of 12 months (range, 12 to 15), and a volumetric calcium score was calculated as an estimate of the total burden of plaque. Treatment with 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors was begun at the discretion of the referring physician. Serial measurements of low-density lipoprotein (LDL) cholesterol were obtained, and the change in the calcium-volume score was correlated with average LDL cholesterol levels. RESULTS: One hundred five patients (70 percent) received treatment with HMG-CoA reductase inhibitors, and 44 patients (30 percent) did not. At follow-up, a net reduction in the calcium-volume score was observed only in the 65 treated patients whose final LDL cholesterol levels were less than 120 mg per deciliter (3.10 mmol per liter) (mean [+/-SD] change in the score, -7+/-23 percent; P=0.01). Untreated patients had an average LDL cholesterol level of at least 120 mg per deciliter and at the time of follow-up had a significant net increase in mean calcium-volume score (mean change, +52+/-36 percent; P<0.001). The 40 treated patients who had average LDL cholesterol levels of at least 120 mg per deciliter had a measurable increase in mean calcium-volume score (25+/-22 percent, P<0.001), although it was smaller than the increase in the untreated patients. CONCLUSIONS: The extent to which the volume of atherosclerotic plaque decreased, stabilized, or increased was directly related to treatment with HMG CoA reductase inhibitors and the resulting serum LDL cholesterol levels. These changes can be determined noninvasively by electron-beam CT and quantified with use of a calcium-volume score. PMID- 9869669 TI - Chemotherapy followed by surgery compared with surgery alone for localized esophageal cancer. AB - BACKGROUND: We performed a multi-institutional randomized trial comparing preoperative chemotherapy followed by surgery with surgery alone for patients with local and operable esophageal cancer. METHODS: Preoperative chemotherapy for patients randomly assigned to the chemotherapy group included three cycles of cisplatin and fluorouracil. Surgery was performed two to four weeks after the completion of the third cycle; patients also received two additional cycles of chemotherapy after the operation. Patients randomly assigned to the immediate surgery group underwent the same surgical procedure. The main end point was overall survival. RESULTS: Of the 440 eligible patients with adequate data , 213 were assigned to receive preoperative chemotherapy and 227 to undergo immediate surgery. After a median possible study time of 55.4 months, there were no significant differences between the two groups in median survival: 14.9 months for the patients who received preoperative chemotherapy and 16.1 months for those who underwent immediate surgery (P=0.53). At one year, the survival rate was 59 percent for those who received chemotherapy and 60 percent for those who had surgery alone; at two years, survival was 35 percent and 37 percent, respectively. The toxic effects of chemotherapy were tolerable, and the addition of chemotherapy did not appear to increase the morbidity or mortality associated with surgery. There were no differences in survival between patients with squamous-cell carcinoma and those with adenocarcinoma. Weight loss was a significant predictor of poor outcome (P=0.03). With the addition of chemotherapy, there was no change in the rate of recurrence at locoregional or distant sites. CONCLUSIONS: Preoperative chemotherapy with a combination of cisplatin and fluorouracil did not improve overall survival among patients with epidermoid cancer or adenocarcinoma of the esophagus. PMID- 9869670 TI - Images in clinical medicine. Malignant hypertension. PMID- 9869671 TI - Attributes of excellent attending-physician role models. AB - BACKGROUND: Although effective role models are important in medical education, little is known about the characteristics of physicians who serve as excellent clinical role models. We therefore conducted a case-control study to identify attributes that distinguish such physicians from their colleagues. METHODS: We asked members of the internal-medicine house staff at four teaching hospitals to name physicians whom they considered to be excellent role models. A total of 165 physicians named by one or more house-staff members were classified as excellent role models (these served as the case physicians in our study). A questionnaire was sent to them as well as to 246 physicians who had residency-level teaching responsibilities but who were not named (controls). Of these 411 physicians, 341 (83 percent) completed questionnaires while unaware of their case-control status. RESULTS: Of the 341 attending physicians who responded, 144 (42 percent) had been identified as excellent role models. Having greater assigned teaching responsibilities was strongly associated with being identified as an excellent role model. In the multivariate analysis, five attributes were independently associated with being named as an excellent role model: spending more than 25 percent of one's time teaching (odds ratio, 5.12; 95 percent confidence interval, 1.81 to 14.47), spending 25 or more hours per week teaching and conducting rounds when serving as an attending physician (odds ratio, 2.48; 95 percent confidence interval, 1.15 to 5.37), stressing the importance of the doctor-patient relationship in one's teaching (odds ratio, 2.58; 95 percent confidence interval, 1.03 to 6.43), teaching the psychosocial aspects of medicine (odds ratio, 2.31; 95 percent confidence interval, 1.23 to 4.35), and having served as a chief resident (odds ratio, 2.07; 95 percent confidence interval, 1.07 to 3.98). CONCLUSIONS: These data suggest that many of the attributes associated with being an excellent role model are related to skills that can be acquired and to modifiable behavior. PMID- 9869672 TI - Creutzfeldt-Jakob disease and related transmissible spongiform encephalopathies. PMID- 9869674 TI - Noninvasive detection of atherosclerosis. PMID- 9869675 TI - Role models--guiding the future of medicine. PMID- 9869676 TI - Self-referral of patients for electron-beam computed tomography to screen for coronary artery disease. AB - As the availability of electron-beam CT increases, it is appropriate to question the balance among medical science, patient care, and profits. Broadening patients' sense of empowerment and promoting their autonomy are worthy goals within medicine. Breast-cancer screening with mammography is an example of a radiographic test used successfully in a diagnostic program based on self referral. But the lessons of such a program, in which the distinction between the disease and the disease-free state is more easily recognized than is the case for age-dependent calcific arterial changes, are not easily extrapolated to screening for coronary disease. Currently, we are facing the possibility that market forces may increase interest in electron-beam CT beyond what is justified by its potential medical benefit. Well-designed clinical trials are required to define fully the appropriate indications for and limitations of electron-beam CT. Such trials will eventually clarify the medical applications of the technique and determine its suitability as a screening procedure for cardiovascular disease. Until then, the use of electron-beam CT, like that of all tests in medicine, should be based on a clearly defined rationale and should be coupled with a medical evaluation by a physician. PMID- 9869677 TI - Change is inevitable PMID- 9869678 TI - Analysis of thoracoscopy in trauma. AB - BACKGROUND: The role of video-assisted thoracic surgery (VATS) in trauma has yet to be established. Up to the time of this writing, reviews of thoracoscopy in trauma have been primarily descriptive rather than analytic. This article analyzes the results of thoracoscopy (nonvideo and VATS) in trauma. METHODS: Analysis was done by reviewing 28 nonoverlapping studies since the introduction of thoracoscopy in 1910, with a combined total of more than 500 patients. RESULTS: Diagnostically, thoracoscopy has been used primarily to evaluate diaphragmatic injury, continued chest tube bleeding, and suspected cardiac injury. Thoracoscopy has a 98% (188/191 patients) accuracy rate in diagnosing diaphragmatic injuries. Therapeutically, thoracoscopy has been used primarily to control chest tube bleeding, evacuate retained hemothoraces, and evacuate empyemas. Thoracoscopy is 90% (89/99 patients) effective in evacuating retained hemothoraces, 86% (19/22 patients) effective in evacuating empyemas, and 82% (33/40 patients) effective in controlling chest tube bleeding. Thoracoscopy benefits include preventing 62% (323/514) of trauma patients from having a thoracotomy or laparotomy. Risks include a 2% (11/534 patients) procedure-related complication rate and a 0.8% (4/471 patients) missed injury rate. Technical failure rates are 10% (10/99 patients) and 4% (7/199 patients) in evacuation of retained hemothoraces and evaluation of diaphragmatic injuries, respectively. CONCLUSIONS: Analysis suggests that thoracoscopy (nonvideo and VATS) can be applied safely and effectively in the care of the injured patient. PMID- 9869679 TI - Adhesion formation is reduced after laparoscopic surgery. AB - BACKGROUND: Adhesion formation after abdominal operations causes significant morbidity. METHODS: Adhesion formation in pigs was compared after placement of prosthetic mesh during celiotomy (group 1), laparoscopy with large incision (group 2), and laparoscopy (group 3). After peritoneum was excised, polypropylene mesh was fixed to the abdominal wall, then to the opposite abdominal wall in the preperitoneal space followed by peritoneal closure. Adhesion area, grade, and vascularity were measured. RESULTS: More adhesions (p < 0.02) covered intraperitoneal mesh (7.57 +/- 1.89 cm2) than covered reperitonealized mesh (2.16 +/- 1.13 cm2), and adhesion grade was significantly greater (p < 0.02). Adhesion areas were significantly greater in groups 1 and 2 than in group 3 (p = 0.001 and 0.03, respectively). Adhesion grade was significantly greater in groups 1 and 2 than in group 3 (p = 0.02 and p = 0.04, respectively). Groups 1 and 2 had more vascular adhesions than group 3 (p < 0.01 and p = 0.02, respectively) CONCLUSIONS: A foreign body within the peritoneum stimulates more numerous and denser adhesions. Tissue trauma distant from the site of adhesions increases their formation. A major advantage of laparoscopic surgery is decreased adhesion formation. PMID- 9869680 TI - Intraabdominal adhesion formation after preperitoneal dissection in the murine model. AB - BACKGROUND: The laparoscopic approach to hernia repair has been advocated by many as a potentially superior method of herniorraphy. Several techniques have been described, each with its own proposed advantages. These techniques involve different anatomic approaches, the most recent of which is the totally extraperitoneal approach (TEPA). One presumed advantage of the extraperitoneal approach is the avoidance of adhesion formation because the peritoneum is not entered and mesh is not placed in direct contact with intra-abdominal structures. We hypothesize, however, that when the peritoneum is dissected from the abdominal wall, it is partially devascularized, leading to scar formation and potential adhesion formation. This would suggest that the TEPA method of herniorraphy may not completely avoid the risks of intra-abdominal adhesion formation. METHODS: After appropriate approval was obtained, 88 male Sprague-Dawley rats were divided into two equal groups. One group underwent laparotomy followed by careful blunt dissection of the peritoneum from the left abdominal wall. The control group underwent laparotomy without manipulation of the peritoneum. All animals were re explored 14 days later, and the abdominal cavity was examined for adhesions. The type and location of any adhesion was recorded. RESULTS: Adhesion formation occurred in 10 of 44 (23%) subjects in the peritoneal dissection group, compared with 3 of 44 (7%) in the nondissection group (p < 0.05). CONCLUSIONS: Dissection of the peritoneum from the overlying abdominal wall in the murine model leads to intra-abdominal adhesion formation. This suggests that peritoneal dissection in the TEPA method of herniorraphy may lead to intra-abdominal adhesion formation. PMID- 9869681 TI - Splenectomy for idiopathic thrombocytopenic purpura: comparison of laparoscopic and conventional surgery. AB - BACKGROUND: This study aimed to compare the safety, efficacy, and clinical benefits of laparoscopic splenectomy (LS) to open splenectomy (OS) in patients with idiopathic thrombocytopenic purpura (ITP). METHODS: The results from 14 consecutive patients who underwent LS for ITP were reviewed and compared with the results from patients who underwent OS for the same disease. Demographics, concomitant disease on admission, and platelet counts were evaluated, as were details of the surgical procedure, postoperative physiologic status, and hospital stay. RESULTS: Mean operative time was 88.3 min for OS and 146.4 min in LS group (p < 0.05). The conversion rate to open splenectomy was 7.1. Therapeutic response to splenectomy was 92.8% in the LS group and 86.6% in the OS group. Bowel canalization, return to liquid diet, and length of hospital stay were all significantly delayed in the OS group as compared with those who underwent LS (p = 0.01, p = 0.02, p = 0.005, respectively). In the OS group the morbidity rate was 13.3%, whereas in the LS group it was 7.1%. CONCLUSIONS: Laparoscopic splenectomy represents a valid alternative to conventional splenectomy in the treatment of ITP. PMID- 9869682 TI - Laparoscopic splenectomy: optimal vascular control using the lateral approach and ultrasonic dissection. AB - BACKGROUND: Intraoperative bleeding is the main complication and main cause of conversion to open surgery during laparoscopic splenectomy (LS). We present the advantages of the lateral approach and the use of the ultrasonic shears (US) for achieving a safer vascular control. METHODS: We have performed a total of 48 LS using several approaches. In our initial experience 10 patients underwent surgery with an anterior approach. In the second series of 18 patients, we used a manually assisted procedure. In the last 20 patients, we employed a technique with a full lateral position. Only three ports have been used. The major part of the dissection was conducted from behind, thus allowing a safer vascular control. The division of short gastric vessels and lower pole vessels was performed using US. The main vascular pedicle was stapled. The spleen was removed through a short Pfannenstiel incision. There were 9 males and 11 females with a mean age of 36 years (range, 18-71 years). Fourteen had immune thrombocytopenic purpura (ITP); two had an HIV-infection-related purpura; two had an autoimmune hemolytic anemia; and two had a spherocytosis. RESULTS: All the 20 LS were completed. The average splenic weight was 274 g (range, 162-1,400 g). In all but one patient, the intraoperative blood loss was less than 60 ml, and was none in six patients (average, 51 ml). In our initial series of 10 patients surgically treated with an anterior approach, the average blood loss was 180 ml, and it was 230 ml in the series of hand-assisted procedures. The average operative time was 127 min (range, 70-220 min), including the time required by the change of position and the Pfannenstiel incision. There was no mortality. All but one patient had an uneventful postoperative course. The HIV-infected patient had a severe postoperative pancreatitis. In those patients with an uncomplicated course, the average postoperative stay was 4.3 days (range, 2-8 days). CONCLUSIONS: The lateral position with a posterior approach to splenic vessels allows for safe vascular control. The use of US results in a gain of time and safety. The average intraoperative bleeding of this series is much lower than that observed in our previous experience and in other published series. PMID- 9869683 TI - Laparoscopic surgery combined with preservation of the spleen for distal pancreatic tumors. AB - BACKGROUND: Laparoscopic distal pancreatectomy combined with spleen salvage by preservation of the splenic vessels has been described in selected patients with islet cell tumors. METHODS: Laparoscopic resection of the left side of the pancreas with spleen preservation on the vasa brevia was attempted in six consecutive patients. RESULTS: Four distal pancreatectomies with spleen preservation were completed laparoscopically. There were two conversions to laparotomy. The median operating time was 300 min (range, 240-360). There was no mortality, but two patients developed a pancreatic fistula. The median postoperative hospital stay was 34.5 days (range, 5-60). All the patients remain well at a median follow-up of 30 months (range, 22-41). CONCLUSIONS: Minimally invasive surgery for distal pancreatic tumors is feasible and appropriate for most benign tumors. The spleen can be safely preserved laparoscopically on its blood supply from the short gastric vessels. The operative technique and especially the closure of the pancreatic stump need further study. PMID- 9869684 TI - Laparoscopic adrenalectomy for nonmalignant disease: improved safety, morbidity, and cost-effectiveness. AB - BACKGROUND: Laparoscopic adrenalectomy has rapidly gained widespread acceptance for treatment of benign adrenal neoplasms. A number of authors have compared various anatomic approaches to laparoscopic adrenalectomy, comparing length of inpatient stay, transfusion requirements, and perioperative complications. Separate studies have found inpatient stay reduced 40-60% with the use of laparoscopic adrenalectomy vs. an open procedure. METHODS: There have been no studies designed specifically to examine and compare perioperative morbidity, length of stay, and patient charges in patients undergoing laparoscopic adrenalectomy. This report examines the Johns Hopkins Hospital experience with laparoscopic adrenalectomy in 22 patients, comparing length of stay, perioperative morbidity, and patient charges. These data are compared with those seen in 17 patients undergoing open adrenalectomy within our institution and 70 patients at all other nonfederal hospitals in the state of Maryland. RESULTS: Outcomes after laparoscopic versus open adrenalectomy were compared. Resumption of diet (1.6 vs. 6.1 days), independent activity (1.6 vs. 7.9 days), inpatient length of stay (1.7 vs. 7.8 days), and total hospital patient charges ($8,698 vs. $12,610) were all significantly reduced in patients undergoing laparoscopic adrenalectomy at our institution. Similar findings were obtained when our data were compared against adrenalectomy performed at other hospitals within the state of Maryland. Length of stay (1.7 vs. 8.9 days) and total hospital patient charges ($8,698 vs. $13,867) were both significantly reduced compared to state-wide data in patients treated with laparoscopic adrenalectomy. CONCLUSIONS: Although a technically challenging procedure, laparoscopic adrenalectomy provides clear advantages over open procedures for the vast majority of adrenal neoplasms. Our data support the conclusion that laparoscopic adrenalectomy should be considered for all patients with benign adrenal neoplasms. PMID- 9869685 TI - Surgical treatment of pheochromocytomas: laparoscopic or conventional? AB - BACKGROUND: The use of minimally invasive techniques in the surgical treatment of pheochromocytoma is controversial because of possible intraoperative excessive hormone release resulting in cardiovascular instabilities. METHODS: Laparoscopic adrenalectomy was performed in nine patients with a total of 10 pheochromocytomas. Conversion was required in two cases. The relevant data were prospectively documented and compared with a historical group of nine patients who had undergone conventional transabdominal adrenalectomy for unilateral pheochromocytoma. RESULTS: The laparoscopic operations lasted significantly longer than the conventional procedures (median 243 min vs. 100 min, p < 0.01). Intraoperative cardiovascular instabilities (tachycardia, hypertension) occurred in seven laparoscopically and eight conventionally treated patients. All were easily controlled. Blood transfusions were necessary in four patients in the conventional and one patient in the laparoscopic group. Postoperative hospital stay and duration of analgetic treatment were significantly shorter after laparoscopic adrenalectomy. CONCLUSIONS: Laparoscopic adrenalectomy is a safe procedure for patients with pheochromocytoma. PMID- 9869686 TI - Laparoscopic adrenalectomy for benign adrenal neoplasms. AB - BACKGROUND: Since first reported in 1992, laparoscopic adrenalectomy has been used to remove a wide variety of adrenal neoplasms. Indications for use of this technique have not been clearly defined, nor has it been demonstrated to be more cost effective than open adrenalectomy. METHODS: A retrospective comparison was made of 19 consecutive laparoscopic and open adrenalectomies performed in patients with benign adrenal neoplasms in a tertiary-care university teaching hospital over a 3-year period. RESULTS: The two groups were well matched for side of tumor and age. Laparoscopic adrenalectomy was completed in 11 of 12 patients in whom it was attempted. The laparoscopic group had significantly smaller tumor size; shorter operative time, postoperative ileus, and postoperative stay; and decreased operative blood loss and postoperative narcotic requirement. There were no significant differences between groups for operating room or hospital charges. CONCLUSIONS: Laparoscopic adrenalectomy is cost effective and should be the preferred treatment for patients with small benign adrenal neoplasms. PMID- 9869687 TI - Endoscopic axillary exploration and sentinel lymphadenectomy. AB - BACKGROUND: Minimally invasive approaches have changed the practice of surgery in several specialties. The purpose of this study was to develop a reproducible endoscopic technique for the evaluation of the axilla in breast cancer patients. METHODS: A total of 23 patients with biopsy-proven breast carcinoma were enrolled. Patients were positioned in the supine position with the ipsilateral arm abducted at 90 degrees. A 1-cm skin incision was made at the superior aspect of the axilla. Dissection was carried bluntly to the lateral border of the pectoralis major. A balloon distention device was inserted into the tract and distended under endoscopic vision to create a working space. Insufflation was initiated up to a pressure of 8 mmHg. A 30 degrees laparoscope was introduced for visualization of axillary contents. One or two additional 5-mm cannulas were placed as needed under direct visualization. Manipulation of axillary contents was performed, and in 19 patients a sentinel node identification technique was applied. RESULTS: In all patients, using insufflation and minimal instrument dissection, the axillary vein, long thoracic, and thoracodorsal nerves were found in their usual anatomical locations. Utilizing blunt and sharp dissection, the axilla was thoroughly inspected, and individual lymph nodes were easily identified and extracted. In 11 of 19 patients, a sentinel node or blue dye was identified using isosulfan blue. There was a procedure concordance of 84%, and there were no complications. CONCLUSIONS: We describe a novel endoscopic technique for the evaluation of the axilla in breast cancer patients. This technique allows (a) creation of a minimally invasive working space within the axilla, (b) recognition of key axillary anatomic landmarks, and (c) instrument manipulation within the axilla to identify and extract lymph nodes, and apply the sentinel node technique. This is the first report of a minimally invasive approach to axillary exploration to employ sentinel lymph node mapping. PMID- 9869688 TI - Percutaneous cholecystostomy and endoscopic cholecystolithotripsy in the management of acute cholecystitis. AB - BACKGROUND: Percutaneous cholecystostomy is a valuable alternative temporary measure for acute cholecystitis in elderly patients with severe underlying cardiopulmonary disease, but the subsequent management of gallbladder calculi is still controversial. METHODS: Eleven patients treated with percutaneous endoscopic cholecystolithotripsy after percutaneous cholecystostomy were evaluated retrospectively. RESULTS: All patients showed clinical improvement after percutaneous cholecystostomy. Tract dilation succeeded in 9 patients. Complete stone clearance was achieved in seven patients over one to four sessions (average, two sessions). Stone extraction could not be completed in two patients because gallbladder access was lost in one patient, and the other refused further procedure. There were three complications, with two biliary fistulas and one major bile leakage leading to emergency cholecystectomy. The duration of the entire procedure ranged from 30 to 126 days (mean, 58 days). During the follow-up (mean 17.2 months), one patient had recurrent cholangitis and the others remained asymptomatic. CONCLUSIONS: Percutaneous cholecystolithotripsy after percutaneous cholecystostomy is a safe alternative in the management of high-risk elderly patients with acute cholecystitis. PMID- 9869689 TI - The outcome of major biliary tract injury with leakage in laparoscopic cholecystectomy. AB - BACKGROUND: Concern has been expressed regarding the increased rates of biliary tract injury (BTI) at laparoscopic cholecystectomy. The aim of the present investigation was to analyze the outcome of laparoscopic biliary tract injury with leakage. METHODS: Sixteen patients having major laparoscopic BTI with leakage were treated. Thirteen of them were referred to our institution for further treatment. The follow-up was complete and focused on clinical outcome and biochemical analysis. RESULTS: Eight BTI were identified at the time of laparoscopic cholecystectomy, and the procedure was converted to a laparotomy. In eight additional patients, BTI was recognized postoperatively. In this group one patient died because of lately diagnosed biliary peritonitis, whereas in the seven surviving patients nine attempts to repair the BTI and eight other interventions were performed. In the conversion group 14 attempts to repair the BTI and 11 other interventions were needed to completely solve the problems. Final restoration of the BTI was done by Roux-en-Y hepaticojejunostomy in 11 patients and suture repair with T-tube drainage of the bile duct in 4. During a median follow-up time of 63 months, three patients suffered from recurrent segmental cholangitis. In the other patients, neither clinical nor biochemical evidence of biliary disease has been found up to this writing. CONCLUSIONS: Laparoscopic BTI has a high morbidity and mortality rate that seems comparable to BTI at open cholecystectomy. The number of attempts to repair the BTI as well as additional interventions is too high, but in this patient series the final outcome seemed to be similar after BTI recognized during and after laparoscopic cholecystectomy. PMID- 9869690 TI - Morbidity in laparoscopic gynecological surgery: results of a prospective single center study. AB - BACKGROUND: We set out to investigate prospectively the morbidity rate for gynecological laparoscopy patients at a tertiary care center. METHODS: We prospectively recorded data on 743 laparoscopic procedures performed between January 1, 1992 and December 31, 1996. The procedures included 36 diagnostic laparoscopies (4.8%), 115 laparoscopies carried out for minor surgical acts (15.4%), 523 for major surgical acts (70.4%), and 69 for advanced surgical acts (9. 4%). A total of 127 patients had a history of prior laparotomy (17%). All those procedures were performed by young senior surgeons. We defined a complication as an event that had modified the usual course of the procedure or of the postoperative period. For statistical analysis, we used the chi-squared test or Fisher's exact test. RESULTS: Complications occurred in 22 cases; the overall complication rate was 2.9% when all events were considered. One complication (injury of the left primitive iliac artery) was related to insertion of the Veress needle (0.13%). A total of 2,578 trocars were inserted, giving rise to 10 complications (1.3%). Three unintended laparotomies were required for bowel or bladder injuries (0.4%). Finally, the introduction of the laparoscope was responsible for 11 complications (1.4%); this figure represents 50% of all the complications of this series. Eight intraoperative complications (1%) occurred during the laparoscopic surgery (seven severe bleedings and one ureter injury, but no intestinal lesions); laparotomy was required in six of these cases. Three complications occurred during the postoperative stage: one granulomatous peritonitis after intraabdominal rupture of a dermoid cyst, one incisional hernia, and a fast-resolving cardiac arrhythmia. CONCLUSIONS: In our experience, operative gynecological laparoscopy is associated with an acceptable morbidity rate. Moreover, about half of the complications occur during the installation of the laparoscopic procedure, underscoring the usefulness of safety rules. PMID- 9869691 TI - Gastroscopic-assisted laparoscopic wedge resection of B-cell gastric mucosa associated lymphoid tissue (MALT) lymphoma. AB - Helicobacter pylori has been implicated as an etiologic agent for B-cell gastric mucosa-associated lymphoid tissue (MALT) lymphoma. This tumor is considered a low grade entity with an associated indolent course. Some instances of tumor regression with adequate antibiotic treatment and irradication of H. pylori have been reported. Gastric malignancy and more specifically, lymphoma, should be considered in all patients with recurrent or nonhealing ulcers. Diagnostic and treatment strategies for these patients remain controversial. Two patients were evaluated, one for recurrent disease, another for a 3-cm nonhealing ulcer. Both patients refused total gastrectomy. Both patients underwent gastroscopy with simultaneous laparoscopic visualization. The mucosal lesions were identified and removed via laparoscopic stapled wedge resection. A combined endoscopic and laparoscopic approach can be considered as an alternative to radical surgery in the treatment strategy of these tumors. Wedge resection allows for staging and thorough pathologic classification. Further studies are necessary to define the natural history and the role of laparo-endoscopy in the diagnosis, staging, and treatment of B-cell gastric MALT lymphoma. PMID- 9869692 TI - Laparoscopic repositioning of an adjustable silicone gastric band for pouch dilatation and stoma obstruction. AB - Pouch dilatation with stoma obstruction is a well-known late complication after adjustable gastric banding operations for morbid obesity. Surgical treatment of this problem usually results in removal of the band, with or without replacement by another, or in repositioning of the band via laparotomy. We present the case of a patient with late pouch dilatation and stoma obstruction after placement of a Laparoscopic Adjustable Gastric Banding system (LAGB-Bioenterics) and in whom the adjustable band was laparoscopically opened, disconnected from the access port, and repositioned more proximally on the stomach. The postoperative course was uneventful. A postoperative radiographic contrast examination showed a correct repositioning of the band. The case demonstrates that the LAGB can be successfully opened and repositioned by a minimal invasive procedure. This is the first time to our knowledge that such a procedure has been reported. PMID- 9869693 TI - The fate of the dropped gallstones during laparoscopic cholecystectomy. AB - Stones can be spilled from the gallbladder during laparoscopic cholecystectomy. These stones can be left in the peritoneal cavity or trapped at the trocar site. The potential late sequel and associated morbidity are not well documented. We reviewed the records of four patients who underwent laparoscopic cholecystectomy at Mount Sinai Medical Center in New York City who suffered from late complications attributed to gallstones left in the peritoneal cavity or abdominal wall. Four patients presented 1-14 months after laparoscopic cholecystectomy with intraabdominal and abdominal wall abscesses. The spillage of gallstones was noticed during the initial operation only in one of the patients. Three patients required laparotomy and open drainage of intraabdominal abscesses with drainage of pus and gallstones after failed attempts at percutaneous drainage. Two patients underwent local exploration of an abdominal wall abscess containing stones. Stones left in the abdominal cavity or trapped in trocar sites after laparoscopic cholecystectomy can cause serious late complications requiring repeated surgical interventions. Every effort should be made in order to avoid spillage of stones during dissection of the gallbladder and cystic duct and during retrieval of the gallbladder through the abdominal wall. PMID- 9869695 TI - Emergency laparoscopic repair of a paraesophageal hernia. AB - A 63-year-old male with a previously documented paraesophageal hernia presented with acute severe epigastric pain and bloating. He was taken urgently to the operating room for laparoscopic exploration. The hernia sac was reduced with difficulty owing to extensive adhesions and the incarcerated portion of the stomach was mottled and blue. After 10 min of observation the stomach began to resume a normal appearance. The anterior crura were approximated and an anterior gastropexy was performed. The patient was discharged on the 3rd postoperative day and has been asymptomatic since. Paraesophageal hernias with evidence of impending gastric necrosis can be approached laparoscopically as long as basic principles are observed. PMID- 9869694 TI - A new technique for laparoscopic resection of a submucosal tumor on the posterior wall of the gastric fundus. AB - Several reports have been published which describe the technique of using an Endo GIA to resect submucosal tumors on the anterior wall of the stomach. Lesions on the posterior wall, however, especially near the esophagocardiac junction (ECJ), are difficult to resect using these reported techniques. This is because the surgeon must divide the omentum and enter the omental bursa in order to use a similar extraluminal technique. Furthermore, special care must be taken to ensure that resections do not involve the ECJ and narrow the esophagus. In order to overcome these difficulties, we have proposed a new technique for the laparoscopic excision of a submucosal tumor located on the posterior wall of the gastric fundus. The principle of this procedure involves the intraluminal resection of the submucosal tumor, including the surrounding stomach wall, using the Endo GIA. This technique is safe, simple, and effective. We believe that we are the first to address the excision of a submucosal lesion by resecting the full thickness of the posterior gastric wall lesion intraluminally. PMID- 9869696 TI - Totally laparoscopic abdominal aortic aneurysm repair. AB - On the basis of our previous animal and clinical experience with laparoscopic intra-abdominal vascular reconstructions, and due to the prevalence of abdominal aortic aneurysms (AAA), we have recently broadened our scope to tackle more difficult aortic surgery laparoscopically. We present a case report of our first clinical experience with laparoscopic AAA repair using specialized laparoscopic vascular instrumentation. The patient was an 84-year-old hypertensive male with a 7-cm asymptomatic infrarenal abdominal aortic aneurysm that was discovered incidentally. He presented with postcoronary artery bypass grafting and had moderate chronic obstructive pulmonary disease (COPD). A spiral computed tomograph (CT) angiogram revealed an adequate infrarenal neck and aneurysmal involvement of the proximal iliac arteries. An eight-port transabdominal technique was used with the patient in the supine position. Proximal and distal control was achieved without difficulty. The aneurysm was excluded using endoscopic stapling devices, and an aortobiiliac reconstruction was performed with a 16 x 9-mm bifurcated dacron graft. Estimated blood loss was 1000 ml, and the operative time was approximately 7 hours. The patient was ambulating without assistance on postoperative day 3. Total hospitalization was 7 days (delayed secondarily to postoperative ileus). Minimal quantities of narcotics were required for analgesia. At 6-months follow-up, the patient has palpable peripheral pulses and no complications related to surgery. This case report shows that a completely laparoscopic approach to the abdominal aortic aneurysm is possible using instrumentation specifically designed for laparoscopic vascular surgery. The exact role that laparoscopic techniques will hold in vascular surgery remains to be determined because these procedures are time consuming and technically difficult. PMID- 9869697 TI - Endoscopic use of fibrin adhesives: problems when injecting through long catheters. AB - When used endoscopically fibrin adhesives are injected through twin-lumen catheters, which can reach up to 180 cm in length. Because fibrinogen solutions have a high viscosity, significant delivery forces are necessary, resulting in discomfort for the operator. Therefore, the two predominant fibrin sealants were characterized with respect to their viscosity and the force needed for their injection. Viscosity was determined at 18 degrees C, 25 degrees C, and 37 degrees C in a micro-Ostwald viscosimeter. Additionally, the maximum forces needed for injection through a 27-cm and a 160-cm catheter were determined at 25 degrees C in an Instron materials testing machine. Compared with preparation A the viscosity of preparation B was 8.0-34.5 times higher at 18 degrees C, 4.6-13.8 times higher at 25 degrees C and 3.1-6.4 times higher at 37 degrees C. In consequence, the delivery forces were 1.5-2.5 times (27 cm probe) and 3.4-4.5 times (160 cm probe) the values determined for preparation A. For preparation B a maximum load of 3.8 kg was necessary for injection. Assuming that different adhesive preparations have the same effect, a preparation of lower viscosity seems to be more suitable for use via long catheters than a preparation of high viscosity. PMID- 9869698 TI - Umbilical one-puncture laparoscopic-assisted appendectomy in children. AB - To perform a laparoscopic appendectomy, three trocars are usually needed. In order to reduce abdominal wall trauma, we have adopted an umbilical one-puncture laparoscopic-assisted appendectomy (UOPLAA). We did a retrospective study of UOPLAA performed during last 2 years on 200 children aged from 5 to 18 years (median, 9 years). The patients were selected after clinical examination. No child with advanced generalized peritonitis or an abscess with a palpable mass was a candidate for this technique. UOPLAA was successful in 184 patients (92%). In 16 cases (8%), an additional trocar was required to manage perforated or retrocecal appendicitis. The mean operative time was 15 min, and the mean hospital stay was 2 days. There were no intraoperative complications. There were 10 (5%) postoperative complications (three parietal and seven intraabdominal). Four patients (2%) needed reoperation under general anesthesia. The UOPLAA is our preference in cases of acute nonperforated appendicitis because it is simple and fast, with good cosmetic results; but in 8% of our cases, an intraoperative difficulty (retrocecal location, abnormal adhesive band, peritonitis, etc.) arose that required the introduction of additional devices to ensure the safety of the laparoscopic procedure. PMID- 9869699 TI - Retroperitoneal approach in laparoscopic adrenalectomy: is it advantageous? AB - Recently, the retroperitoneal laparoscopic approach has been described as advantageous in avoiding the respiratory and hemodynamic effects of CO2 pneumoperitoneum and giving direct access without the need to move abdominal organs. Forty-two laparoscopic adrenalectomies (LpA) were performed in 36 patients with a variety of adrenal disorders, including 9 patients with nonfunctioning tumors, 11 patients with aldosterone adenoma, 10 patients with Cushing's adenoma, and 6 patients with Cushing's disease. Twenty-two adrenalectomies were performed using the transperitoneal approach (TLpA), and 20 via the retroperitoneoscopic approach (RLpA). Arterial blood samples, mean arterial pressure, heart rate, and clinical parameters were evaluated. At the end of the operation, the PaCO2, PetCO2, and base deficit all increased significantly in both retroperitoneal and transperitoneal CO2 insufflation compared with basal values. Arterial pH decreased significantly in both TLpA and RLpA groups. All clinical parameters evaluated (operation time, analgesic dosing requirements, hospital stay, and the days until return to normal activity) were similar in the TLpA and RLpA approaches. Two patients in the TLpA (10.5%) group and two patients in the RLpA (10%) group needed conversion to open surgery. This study shows the safety and efficacy of laparoscopic adrenalectomy via the transperitoneal or retroperitoneal route in patients with a variety of adrenal disorders. The retroperitoneoscopic approach could be the primary choice in patients with previous abdominal surgery. PMID- 9869700 TI - Usefulness of a visceral mini-retractor accessible without trocar port during laparoscopic surgery. PMID- 9869701 TI - Tailored augmentation of the lower esophageal sphincter in experimental antireflux operations. PMID- 9869703 TI - Gasless videoendoscopic implantation of aortobifemoral vascular prostheses via transperitoneal or extraperitoneal approach in an animal model. PMID- 9869704 TI - News and notices PMID- 9869705 TI - Neuroanatomical correlates of human reasoning. AB - One of the important questions cognitive theories of reasoning must address is whether logical reasoning is inherently sentential or spatial. A sentential model would exploit nonspatial (linguistic) properties of representations whereas a spatial model would exploit spatial properties of representations. In general terms, the linguistic hypothesis predicts that the language processing regions underwrite human reasoning processes, and the spatial hypothesis suggests that the neural structures for perception and motor control contribute the basic representational building blocks used for high-level logical and linguistic reasoning. We carried out a [(15)O] H(2)O PET imaging study to address this issue. Twelve normal volunteers performed three types of deductive reasoning tasks (categorical syllogisms, three-term spatial relational items, and three term nonspatial relational items) while their regional cerebral blood flow pattern was recorded using [(15)O] H(2)O PET imaging. In the control condition subjects semantically comprehended sets of three sentences. In the deductive reasoning conditions subjects determined whether the third sentence was entailed by the first two sentences. The areas of activation in each reasoning condition were confined to the left hemisphere and were similar to each other and to activation reported in previous studies. They included the left inferior frontal gyrus (Brodmann area 45, 47), a portion of the left middle frontal gyrus (Brodmann area 46), the left middle temporal gyrus (Brodmann areas 21, 22), a region of the left lateral inferior temporal gyrus and superior temporal gyrus (Brodmann areas 22, 37), and a portion of the left cingulate gyrus (Brodmann areas 32, 24). There was no significant right-hemisphere or parietal activation. These results are consistent with previous neuroimaging studies and raise questions about the level of involvement of classic spatial regions in reasoning about linguistically presented spatial relations. PMID- 9869706 TI - A functional neuroimaging description of two deep dyslexic patients. AB - Deep dyslexia is a striking reading disorder that results from left-hemisphere brain damage and is characterized by semantic errors in reading single words aloud (e.g., reading 'spirit' as 'whisky'). Two types of explanation for this syndrome have been advanced. One is that deep dyslexia results from a residual left-hemisphere reading system that has lost the ability to pronounce a printed word without reference to meaning. The second is that deep dyslexia reflects right-hemisphere word processing. Although previous attempts to adjudicate between these hypotheses have been inconclusive, the controversy can now be addressed by mapping functional anatomy. In this study, we demonstrate that reading by two deep dyslexic patients (CJ and JG) involves normal or enhanced activity in spared left-hemisphere regions associated with naming (Broca's area and the left posterior inferior temporal cortex) and with the meanings of words (the left posterior temporo-parietal cortex and the left anterior temporal cortex). In the right-hemisphere homologues of these regions, there was inconsistent activation within the normal group and between the deep dyslexic patients. One (CJ) showed enhanced activity (relative to the normals) in the right anterior inferior temporal cortex, the other (JG) in the right Broca's area, and both in the right frontal operculum. Although these differential right hemisphere activations may have influenced the reading behavior of the patients, their activation patterns primarily reflect semantic and phonological systems in spared regions of the left hemisphere. These results preclude an explanation of deep dyslexia in terms of purely right-hemisphere word processing. PMID- 9869707 TI - Coupled temporal memories in Parkinson's disease: a dopamine-related dysfunction. AB - Dysfunction of the basal ganglia and the brain nuclei interconnected with them leads to disturbances of movement and cognition, including disordered timing of movement and perceptual timing deficits. Patients with Parkinson's disease (PD) were studied in temporal reproduction tasks. We examined PD patients when brain dopamine (DA) transmission was impaired (OFF state) and when DA transmission was reestablished, at the time of maximal clinical benefit following administration of levodopa + apomorphine (ON state). Patients reproduced target times of 8 and 21 sec trained in blocked trials with the peak interval procedure, which were veridical in the ON state, comparable to normative performance by healthy young and aged controls (Experiment 1). In the OFF state, temporal reproduction was impaired in both accuracy and precision (variance). The 8-sec signal was reproduced as longer and the 21-sec signal was reproduced as shorter than they actually were (Experiment 1). This "migration" effect was dependent upon training of two different durations. When PD patients were trained on 21 sec only (Experiment 2), they showed a reproduction error in the long direction, opposite to the error produced under the dual training condition of Experiment 1. The results are discussed as a mutual attraction between temporal processing systems, in memory and clock stages, when dopaminergic regulation in the striatum is dysfunctional. PMID- 9869708 TI - Perseveration and strategy in a novel spatial self-ordered sequencing task for nonhuman primates: effects of excitotoxic lesions and dopamine depletions of the prefrontal cortex. AB - Damage to the prefrontal cortex disrupts the performance of self-ordered sequencing tasks, although the precise mechanisms by which this effect occurs is unclear. Active working memory, inhibitory control, and the ability to generate and perform a sequence of responses are all putative cognitive abilities that may be responsible for the impaired performance that results from disruption of prefrontal processing. In addition, the neurochemical substrates underlying prefrontal cognitive function are not well understood, although active working memory appears to depend upon an intact mesocortical dopamine system. The present experiments were therefore designed to evaluate explicitly the contribution of each of these abilities to successful performance of a novel spatial self-ordered sequencing task and to examine the contribution of the prefrontal cortex and its dopamine innervation to each ability in turn. Excitotoxic lesions of the prefrontal cortex of the common marmoset profoundly impaired the performance of the self-ordered sequencing task and induced robust perseverative responding. Task manipulations that precluded perseveration ameliorated the effect of this lesion and revealed that the ability to generate and perform sequences of responses was unaffected by excitotoxic damage to prefrontal cortex. In contrast, large dopamine and noradrenaline depletions within the same areas of prefrontal cortex had no effect on any aspect of the self-ordered task but did impair the acquisition of an active working memory task, spatial delayed response, to the same degree as the excitotoxic lesion. These results demonstrate that a lesion of the ascending monoamine projections to the prefrontal cortex is not always synonymous with a lesion of the prefrontal cortex itself and thereby challenge existing concepts concerning the neuromodulation of prefrontal cognitive function. PMID- 9869709 TI - Is the category-specific deficit for living things spurious? AB - Some neurological patients make more errors naming living than nonliving things. This is true even when the difficulty of naming living and nonliving things, as measured by the performance of normal subjects, is taken into account. Gaffan and Keywood (1993) argued that this apparently selective deficit for living things is spurious and offered an alternative explanation of it in terms of the effect of noise on regression analyses. We report Monte Carlo simulations showing that, in two patients, the living things deficit cannot be accounted for this in this year. These simulations are more consistent with the claim that the recognition of living things depends on specialized mechanisms. PMID- 9869710 TI - Semantic and visual determinants of face recognition in a prosopagnosic patient. AB - Prosopagnosia is the neuropathological inability to recognize familiar people by their faces. It can occur in isolation or can coincide with recognition deficits for other nonface objects. Often, patients whose prosopagnosia is accompanied by object recognition difficulties have more trouble identifying certain categories of objects relative to others. In previous research, we demonstrated that objects that shared multiple visual features and were semantically close posed severe recognition difficulties for a patient with temporal lobe damage. We now demonstrate that this patient's face recognition is constrained by these same parameters. The prosopagnosic patient ELM had difficulties pairing faces to names when the faces shared visual features and the names were semantically related (e.g., Tonya Harding, Nancy Kerrigan, and Josee Chouinard -three ice skaters). He made tenfold fewer errors when the exact same faces were associated with semantically unrelated people (e.g., singer Celine Dion, actress Betty Grable, and First Lady Hillary Clinton). We conclude that prosopagnosia and co-occurring category-specific recognition problems both stem from difficulties disambiguating the stored representations of objects that share multiple visual features and refer to semantically close identities or concepts. PMID- 9869711 TI - H.M. revisited: relations between language comprehension, memory, and the hippocampal system. AB - Three studies tested the claim that H.M. exhibits a "pure memory deficit" that has left his ability to comprehend language unimpaired relative to memory-normal controls. In Study 1, H.M. and memory-normal controls of comparable intelligence, education, and age indicated whether sentences were ambiguous or unambiguous, and H. M. detected ambiguities significantly less often than controls. In Study 2, participants identified the two meanings of visually presented sentences that they knew were ambiguous, and relative to controls, H.M. rarely discovered the ambiguities without help and had difficulty understanding the first meanings, experimenter requests, and his own output. Study 3 replicated these results and showed that they were not due to brain damage per se or to cohort effects: Unlike H.M., a patient with bilateral frontal lobe damage detected the ambiguities as readily as young and same-cohort older controls. These results bear on two general classes of theories in use within a wide range of neurosciences and cognitive sciences: The data favor "distributed-memory theories" that ascribe H.M.'s deficit to semantic-level binding processes that are inherent to both language comprehension and memory, over "stages-of-processing theories," where H.M.'s defective storage processes have no effect on language comprehension. PMID- 9869712 TI - Deficits in complex visual perception following unilateral temporal lobectomy. AB - Although human temporal cortex is known to be important for short- and long-term memory, its role in visual perception is not well understood. In this study, we compared the performance of three patients with unilateral temporal lobectomies to that of normal controls on both "simple" and "complex" visual discriminations that did not involve explicit memory components. Two types of complex tasks were tested that involved discriminations secondary to texture segmentation. These were contrasted with simple discriminations using luminance-defined stimuli. Patients showed impaired thresholds only on tasks involving texture segmentation, performing as well as controls when the targets were defined by luminance rather than texture. The minimum stimulus presentation times for threshold performance were also measured for all tasks and found to be elevated in temporal lobectomy patients relative to controls. Although the magnitude of the deficits observed was substantial, loss was equivalent in ipsi- and contra-lesional regions of the visual field. Additional control experiments showed that the patients' perceptual deficits were not due, even in part, to disturbances of basic visual capacities such as acuity and contrast sensitivity. Our results indicate that temporal lobe damage disrupts complex, but not simple, visual discriminations throughout the visual field. PMID- 9869713 TI - Semantic processing of spoken words in Alzheimer's disease: an electrophysiological study. AB - Patients suffering from Alzheimer's disease (AD) have severe difficulties in tasks requiring the use of semantic knowledge. The semantic deficits associated with AD have been extensively studied by using behavioral methods. Many of these studies indicate that AD patients have a general deficit in voluntary access to semantic representations but that the structure of the representations themselves might be preserved. However, several studies also provide evidence that to some extent semantic representations in AD may in fact be degraded. Recently, a few studies have utilized event-related brain potentials (ERPs) that are sensitive to semantic factors in order to investigate the electrophysiological correlates of the semantic impairment in AD. Interest has focused on the N400 component, which is known to reflect the on-line semantic processing of linguistic and pictorial stimuli. The results from studies of N400 changes in AD remain somewhat controversial: Some studies report normal or enlarged N400 components in AD, whereas others report diminished ones. One issue not reported in previous studies is whether word-elicited ERPs other than N400 remain normal in AD. In the present study our aim was to find out whether the ERP waveforms N1, P2, N400, and Late Positive Component (LPC) to semantically congruous and incongruous spoken words are abnormal in AD and whether such abnormalities specifically reflect deficiencies in semantic activation in AD. Auditory ERPs from 20 scalp sites to semantically congruous and incongruous final words in spoken sentences were recorded from 17 healthy elderly adults and 9 AD patients. The early ERP waveforms N1 and P2 were relatively normal for the AD patients, but the N400 and LPC effects (amplitude difference between congruous and incongruous conditions) were significantly reduced. We interpret the present results as showing that semantic-conceptual activation and other high-level integration processes are defective in AD. However, a word congruity effect earlier than N400 (phonological mismatch negativity), reflecting lexical selection processes, is at least to some extent preserved in AD. PMID- 9869714 TI - Auxiliary selection in Italian: a comment on Miozzo and Caramazza's "On knowing the auxiliary of a verb that cannot be named: evidence for the independence of grammatical and phonological aspects of lexical knowledge". PMID- 9869715 TI - Photodynamic therapy for Barrett's esophagus: follow-up in 100 patients. AB - BACKGROUND: This report presents clinical results of photodynamic therapy in patients with Barrett's esophagus and dysplasia or superficial esophageal cancer. METHODS: One hundred patients including 13 with superficial cancers were treated. Light (630 nm) was endoscopically delivered to the esophageal mucosa by a diffuser or a windowed esophageal centering balloon. Nd:YAG laser was required to ablate small residual areas of Barrett's mucosa during-long-term follow-up. Patients were maintained on omeprazole and were followed for 4 to 84 months (mean 19 months). RESULTS: Conversion of approximately 75% to 80% of treated Barrett's mucosa to normal squamous epithelium was found in all patients; complete elimination of Barrett's mucosa was noted in 43 patients. Dysplasia was eliminated in 78 patients. Dysplasia developed during follow-up in 11 of 48 patients in untreated Barrett's mucosa requiring additional therapy. Ten of the 13 malignancies were ablated. Esophageal strictures occurred in 34%. Use of longer centering balloons reduced the incidence of strictures. CONCLUSION: Photodynamic therapy alone or with Nd:YAG laser thermal ablation combined with long-term acid inhibition provides an effective endoscopic therapy to (1) eliminate Barrett's mucosal dysplasia and superficial esophageal cancer and (2) reduce the extent of and, in some cases, eliminate Barrett's mucosa. PMID- 9869716 TI - KTP laser destruction of dysplasia and early cancer in columnar-lined Barrett's esophagus. AB - BACKGROUND: The rising incidence of esophageal adenocarcinoma in western countries requires a new strategy in the management of dysplasia in Barrett's esophagus. Esophagectomy, which has high morbidity and mortality rates, has been recommended to treat patients with severe dysplasia. Strictly superficial laser coagulation with tissue ablation therefore is a desirable option for the management of dysplasia in Barrett's esophagus because the tissue to be ablated is only about 2 mm thick. Potassium-titanyl-phosphate (KTP) laser light with a wavelength of 532 nm is preferentially absorbed by hemoglobin and therefore combines excellent coagulation with limited tissue penetration. We report first clinical results with KTP laser superficial vaporization of dysplasia and early cancer in Barrett's esophagus. METHODS: Eight men and 2 women 43 to 84 years of age with short segments of Barrett's esophagus or traditional Barrett's esophagus and histologically proved low-grade (n = 4) and high-grade (n = 4) dysplasia or early adenocarcinoma (n = 2) were selected for this pilot study. For all patients thermal endoscopic destruction was conducted with a frequency-doubled neodymium:yttrium-aluminum-garnet (Nd:YAG) KTP laser system. Laser therapy was performed by means of the free-beam method with coaxial insufflation of gas. An average of 2.4 sessions per patient were required for ablation of the Barrett's mucosa. RESULTS: Two to three days after laser treatment the response of the ablated mucosa was assessed with endoscopy and biopsy. Samples taken showed fibrinoid necrosis of the mucosal layer. A complete response was obtained for all 10 patients. Replacement by normal squamous cell epithelium was induced in combination with acid suppression therapy of up to 80 mg omeprazole daily. No complications occurred. In two patients biopsy showed specialized mucosa beneath the restored squamous cell epithelial layer. Follow-up times were as long as 15 months (mean value 10.6 months). CONCLUSIONS: KTP laser destruction of Barrett's esophagus induced mucosal regeneration with normal squamous cell epithelium in combination with acid suppression. Limitation of the depth of thermal destruction in Barrett's esophagus minimizes risk for perforation or stricture formation. KTP laser ablation of Barrett's esophagus seems to be feasible and safe in short segments of Barrett's esophagus with dysplasia or early cancer. PMID- 9869717 TI - Comparison of the hemostatic efficacy of the endoscopic hemoclip method with hypertonic saline-epinephrine injection and a combination of the two for the management of bleeding peptic ulcers. AB - BACKGROUND: The endoscopic hemoclip method is a safe and effective hemostatic method for managing bleeding peptic ulcers. We compared the hemostatic efficacy of the endoscopic hemoclip method with that of hypertonic saline-epinephrine (HSE) injection and a combined method in the management of bleeding peptic ulcers. METHODS: From July 1994 to July 1997, we conducted a randomized clinical trial of endoscopic hemostasis involving 124 patients with actively bleeding or visible vessels at endoscopic inspection. RESULTS: Patients were randomly assigned to hemoclip (41 patients), HSE (41 patients), and combined treatment groups (42 patients). Initial hemostasis was achieved in 97.6%, 95.1%, and 97.6% of cases, respectively. Recurrent bleeding developed in 2.4%, 14.6%, and 9.5% of cases. Emergency operations were performed in 4.9%, 14.6%, and 2.3% of cases. The hemostasis rate was 71.4%, 50%, and 66.7% for spurting hemorrhage in each group. Permanent hemostasis was achieved in 95.1%, 85.4%, and 95.2% of cases. Three patients had complications, all in the HSE group. CONCLUSIONS: The hemoclip method is an effective hemostatic procedure and is safer than HSE injection. The combined method does not provide substantial advantage over use of the hemoclip method alone in the hemostatic management of bleeding peptic ulcers. PMID- 9869718 TI - Cost effectiveness of EUS for preoperative localization of pancreatic endocrine tumors. AB - BACKGROUND: Endoscopic ultrasonography (EUS) is highly accurate in the localization of small pancreatic tumors. We determined the cost effectiveness of EUS used early in the preoperative evaluation of pancreatic endocrine tumors. METHODS: In a study with a case-control design, 36 patients (19 men, 17 women) who underwent preoperative EUS were matched retrospectively with 36 patients who underwent surgical exploration immediately before the introduction of EUS. The number, cost, and effectiveness of preoperative localization studies, days of hospitalization, and surgical and anesthesia times were assessed. RESULTS: The EUS group had reduced charges for preoperative localization studies: $2620 versus $4846 per patient (p < 0.05), largely because of reductions in the number of diagnostic angiograms and venous sampling procedures performed. Surgical and total anesthesia times were decreased, as were the number of preoperative admissions for angiographic procedures. The cost-effectiveness ratio for the EUS group was $3144 per tumor localized compared with $5628 per tumor localized for the group treated before EUS became available (p < 0.05). CONCLUSION: EUS is highly accurate in the localization of pancreatic neuroendocrine tumors and is cost effective when used early in the preoperative localization strategy. EUS decreased the need for additional invasive tests and avoided unnecessary morbidity and resource consumption. EUS should play a primary role in preoperative localization of pancreatic neuroendocrine tumors. PMID- 9869719 TI - Diagnosis of choledocholithiasis: EUS or magnetic resonance cholangiography? A prospective controlled study. AB - BACKGROUND: Endoscopic ultrasonography (EUS) appears to be the best imaging method for the diagnosis of choledocholithiasis. The aim of this preliminary, prospective, controlled study was to assess the accuracy of EUS and magnetic resonance cholangiopancreatography (MRCP) in the diagnosis of common bile duct stones. METHODS: From December 1995 through April 1997, all patients referred because of suspicion of the presence of common bile duct stones were included in the study. EUS and MRCP were performed. Each examination was performed by a different operator unaware of the result of the other procedure. The definitive diagnosis was established by means of endoscopic retrograde cholangiography with sphincterotomy or a surgical procedure. RESULTS: Forty-three patients (18 men, 25 women) with a mean age of 60.9 +/- 14.5 years (range 25 to 81 years) were included in the study. Eleven patients were excluded because of unavailability of magnetic resonance imaging(n = 5) or EUS (n = 6). Ten patients (31.2%) had choledocholithiasis. For this diagnosis, the sensitivity of EUS was 100%, the specificity was 95.4%, the positive predictive value was 90.9%, and the negative predictive value was 100%. The corresponding values for MRCP were 100%, 72.7%, 62.5%, and 100%, not significantly different from EUS results. The accuracy of EUS was 96.9%, and that of MRCP was 82.2%. CONCLUSION: This preliminary study confirmed EUS as an accurate and noninvasive procedure for the diagnosis of common bile duct stones. MRCP, which had a high sensitivity and high negative predictive value, might be an accurate technique for patients with a contraindication to EUS. PMID- 9869720 TI - Endoscopic urease sensor system for detecting Helicobacter pylori on gastric mucosa. AB - BACKGROUND: It is now accepted that the curing of Helicobacter pylori infection will result in healing of chronic active gastritis and will change the natural history of gastroduodenal ulcer disease. Both endoscopic observation and evaluation of H. pylori status of the stomach are necessary for diagnosis and treatment of such patients. We carried out a clinical evaluation of an endoscopic tube type urease sensor system for the detection of H. pylori on the gastric mucosa. The differential output of two pH-sensitive field effect transistors at the tip of the endoscopic tube reflects the pH change in a urea solution depending on the existence of urease. METHODS: In vitro experiments and clinical evaluation of the system were performed. Fifty-one patients who were suspected to have a gastroduodenal disorder were examined for H. pylori infection with this system, using the combination of histologic and bacteriologic examinations and rapid urease test as the references. RESULTS: Clinical sensitivity and specificity of this system were 26 of 28 (92.9%) and 22 of 23 (95.7%), respectively. A measurement at 1 site is completed in about 1 minute. Repetition of the procedure provides multi-site measurements. CONCLUSIONS: The present system makes possible quick on-site detection of H. pylori under endoscopic observation, with satisfactory sensitivity and specificity. PMID- 9869721 TI - Reuse of disposable sphincterotomes for diagnostic and therapeutic ERCP: a one year prospective study. AB - BACKGROUND: Although previous studies have documented that single-channel sphincterotomes can be sterilized and used up to 10 times, no studies have prospectively evaluated the reuse of disposable sphincterotomes. METHODS: Five- and six-French double-channel sphincterotomes marketed as one-time-use only were manually cleaned, sterilized with ethylene oxide, and used up to 10 times. Over a 1-year period, total number of uses as a cannulation or cutting device, reason for sphincterotome dysfunction, possible infectious complications were evaluated, and cost-savings associated with reuse were evaluated. RESULTS: The mean number of times sphincterotomes could be used in our institution was 3.4; a broken or stiff cutting wire was the main reason for discard. There were two infectious complications during the study period. Both were experienced by patients with unrelieved obstruction treated with new sphincterotomes. Contingent on the method used, total savings related to sphincterotome reuse was approximately $66,000. CONCLUSIONS: Double-channel sphincterotomes marketed as one-time-use items can be reused safely when properly cleaned. Reuse was associated with significant cost savings in our institution. PMID- 9869722 TI - Electrophysical factors influencing endoscopic sphincterotomy. AB - BACKGROUND: Analog computer techniques were used to measure electrosurgical power during sphincterotomy in experimental models and patients. METHODS: Total energy and transient changes in power were measured during sphincterotomy of bile ducts in the livers of pigs, ampullae of humans post mortem, and during clinical sphincterotomy. The effect of waveform on hemostasis was studied in experiments on canine mesenteric arteries. RESULTS: Electrosurgical waveforms (CUT, COAG, BLEND) were measured. Halving wire contact length halved energy needed to initiate cutting. The CUT waveform rarely initiated cutting at lower power settings than the BLEND waveform. With CUT, BLEND, and COAG waveforms, approximately the same energy initiated cutting. Efficiency of cutting increased linearly with power. The COAG waveform required higher power settings than BLEND or CUT to initiate cutting (p < 0.05). Force and wire diameter influenced cutting. BLEND was more effectively hemostatic than CUT (p < 0.05). COAG was significantly more hemostatic than BLEND and CUT. Cutting efficiency during clinical sphincterotomy was poor. CONCLUSIONS: This work has practical implications. Shortening wire contact length was effective in starting a cut at suboptimal settings, whereas changing from BLEND to CUT made little difference. Increasing power setting may help if cutting does not start. BLEND stops bleeding better than CUT. COAG stops bleeding better than BLEND but cuts poorly. Cutting during clinical sphincterotomy is inefficient and can be improved. PMID- 9869723 TI - Clinical presentation and short-term outcome of endoscopic therapy of patients with symptomatic incomplete pancreas divisum. AB - BACKGROUND: The clinical significance of incomplete pancreas divisum (IPD) has not been fully described. In this study we report the clinical presentation and results of endoscopic treatment of the 32 (0.6%) patients with IPD seen at our center over a 10-year period. METHODS: The study population consisted of 24 women and 8 men (mean age 42 years, range 13 to 82 years). Ten (31%) patients presented with acute recurrent pancreatitis, 5 (16%) with chronic pancreatitis, and 3 (9%) with pancreatic type pain. Detailed history, laboratory tests, US, CT, and ERCP excluded other etiologies for their symptoms. The remaining 14 (44%) presented with biliary problems. The 18 symptomatic patients with IPD were treated as follows: 8 received dorsal duct stents, 3 underwent minor papilla endoscopic sphincterotomy and dorsal duct stent placement, 4 had minor papilla dilatation only, and 3 had ventral duct stents placed. RESULTS: Patients were then followed for recurrence of pancreatitis and pancreatic-type pain. Mean follow-up was 15.5 months (range 3 to 30 months). Six (60%) of the patients with acute recurrent pancreatitis and 4 (80%) with chronic pancreatitis benefitted from the endoscopic therapy. However, only 1 (33%) of the patients with pancreatic-type pain benefitted. CONCLUSION: The clinical presentation and response to endoscopic therapy of patients with ICP appeared to be similar to that of patients with complete pancreas divisum. PMID- 9869724 TI - Success of repeat ERCP by the same endoscopist. AB - BACKGROUND: The success rate of repeat endoscopic retrograde cholangiopancreatography (ERCP) by the same endoscopist after a failed initial attempt is unknown. It has been suggested that a repeat examination at a tertiary referral center be performed after an unsuccessful attempt. Our aim was to determine the success rate of repeat ERCP at a different endoscopic session by the same endoscopist and the outcomes among patients with a failed index procedure. METHODS: A review of 500 consecutive ERCP procedures was performed at a teaching institution. RESULTS: The overall initial success rate for cannulation of the duct of interest was 90.8% at index endoscopy. Endoscopy was repeated after 51% unsuccessful procedures, and access to the desired duct was achieved in 87.5% of repeat attempts. A needle knife was used in 21% instances, and its use facilitated cannulation of the duct of interest in 80%. No complications occurred with repeat ERCP. Of the 3 patients who underwent failed repeated ERCP, 1 was not available for the follow-up study, 1 had metastatic cancer, and the other had pancreas divisum. The outcomes among patients who did not undergo repeat ERCP included malignant disease diagnosed with other imaging techniques (35% of patients) and no further admissions or emergency room visits for suspected pancreaticobiliary symptoms (39% of patients). CONCLUSIONS: Repeat ERCP by the same endoscopist yields an 87.5% success rate. This leads to an overall success rate of 95%. PMID- 9869725 TI - Videoendoscopic diagnosis of esophageal motility disorders. AB - BACKGROUND: Esophageal motility disorders are usually diagnosed by manometry. We evaluated videoendoscopy as a diagnostic test. METHODS: In this study, 20 patients with achalasia, 13 with scleroderma, and 33 control subjects had a standard endoscopic examination followed by protocol videotaping of swallows to observe contractions in the esophagus and in the lower esophageal sphincter. Tapes were later reviewed by 2 blinded observers who recorded their motility findings and diagnoses. RESULTS: In the mid esophagus at 25 cm, lumen-occluding peristaltic contractions were identified in 26 of 33 control subjects versus 1 of 20 achalasia (p < 0.001) and 3 of 13 scleroderma patients (p < 0.005). As viewed in the lower esophagus, the lower esophageal sphincter opened normally in 31 of 33 control subjects versus 1 of 20 achalasia (p < 0.001). In scleroderma, the sphincter never closed in 12 of 13 patients (p < 0. 001 versus control subjects). A diagnostic sequence of sphincter opening followed by contraction in the esophageal body and subsequent sphincter closing was seen in 33 of 33 control subjects, 2 of 20 achalasia, and 1 of 13 scleroderma patients (both, p < 0. 001). The observers made the correct diagnosis in 96% of cases. CONCLUSIONS: Achalasia and esophageal scleroderma can be identified by endoscopic observation of motility. This procedure may represent an adjunctive diagnostic test to manometry. PMID- 9869726 TI - Esophageal fistula sealing: choice of stent, practical management, and cost. AB - BACKGROUND: Three models of covered metal stents are available to seal esophageal fistulas. METHODS: Stainless steel covered stents were inserted in 5 patients (group I); nitinol covered stents were inserted in 12 patients (group II) with malignant (n = 14) or benign (n = 3) esophageal fistulas. RESULTS: Stent positioning was satisfactory in all cases. Fistula sealing was complete in 1 of 5 (20%) and 12 of 12 (100%) patients of groups I and II, respectively (p < 0.005). Continued esophageal leakage was initially related to the passage of fluids alongside the stent covering (n = 3) and to early stent migration (n = 1). Complications related to stent placement were observed in 2 of 17 (12%) patients and were fatal. During follow-up (mean 153 +/- 143 days), esophageal fistulas relapsed after initial sealing in 5 of 13 (38%) patients. Further treatment (glue or fibrin sealant injection, additional stent insertion) was attempted in 7 cases of persistent or relapsing esophageal fistula, with sealing obtained in 5 of them. The costs per patient and per day free from symptoms due to the esophageal fistula were $106 and $57 in groups I and II, respectively. CONCLUSION: Nitinol covered stents more frequently provided complete esophageal fistula sealing, as compared with stainless steel covered stents. Further treatments tailored to the mechanisms of fistula persistence or relapse often provided sealing. PMID- 9869727 TI - Ability of naloxone to enhance the colonoscopic appearance of normal colon vasculature and colon vascular ectasias. AB - BACKGROUND: Colon vascular ectasias are a common cause of lower intestinal bleeding among the elderly. The lesions may be difficult to diagnose at colonoscopy because they are small and their appearance may be influenced by the patient's blood pressure, blood volume, and narcotic sedation during the procedure. The purpose of this study was to determine whether naloxone influenced the appearance of colon vascular ectasias at colonoscopy. METHODS: One hundred forty-four patients older than 60 years undergoing complete colonoscopy participated in the study. Medications were given in the usual doses. After a 2 minute inspection of the cecum and ascending colon, naloxone was given, followed by another 2-minute observation period. Photographic documentation of areas of interest was obtained before and after administration of naloxone. RESULTS: One hundred fourteen patients (79%) had no ectasias before or after administration of naloxone. Fourteen (9.7%) initially had normal vessels, and the vessels became more prominent; 4 (2.7%) initially had no ectasias, but ectasias later developed. Four patients (2.7%) had ectasias before administration of naloxone that did not change; 8 (5.4%) had ectasias before administration of naloxone that increased in size (3 patients), number (7 patients), or both (2 patients). CONCLUSIONS: Naloxone can enhance the appearance of normal colonic vasculature and ectasias. Naloxone is an important adjunctive medication for patients undergoing examinations for lower intestinal bleeding. PMID- 9869728 TI - The role of intra-abdominal pressure on splanchnic and pulmonary hemodynamic and metabolic changes during carbon dioxide pneumoperitoneum. AB - BACKGROUND AND METHODS: To find an intra-abdominal pressure (IAP) range for laparoscopic procedures that elicits only moderate splanchnic and pulmonary hemodynamic and metabolic changes, including hepatic and intestinal tissue pH and superficial hepatic blood flow, we installed an IAP of 7 and 14 mm Hg each for 30 minutes in 10 healthy pigs (30 +/- 4 kg). RESULTS: In parallel with the increase of IAP, the mean transmural pulmonary artery pressure increased (from 25 +/- 3 to 27 +/- 4 at 7 mm Hg IAP and 30 +/- 6 mm Hg at 14 mm Hg IAP, p < 0.05); the pulmonary artery-to-pulmonary capillary wedge pressure gradient also increased (from 17 +/- 2.7 to 21 +/- 3 mm Hg at 7 mm Hg IAP and 24 +/- 4.2 mm Hg at 14 mm Hg IAP, p < 0.01), and the arterial oxygenation decreased (p < 0.005). Relevant changes at an IAP of 14 mm Hg were observed in right atrial pressure during inspiration (from 7 +/- 2 to 12 +/- 3 mm Hg, p < 0. 0001) and in abdominal aortic flow (from 1.43 +/- 0.4 to 1.19 +/- 0. 3 L/min, p < 0.01). However, transmural right atrial pressure and cardiac output remained essentially unchanged. Portal and hepatic venous pressure increased in parallel with the IAP (portal: from 12 +/- 3 to 17 +/- 3 at 7 mm Hg IAP and 22 +/- 3 mm Hg at 14 mm Hg IAP, p < 0.01; hepatic venous: from 8 +/- 3 to 14 +/- 6 at 7 mm Hg IAP and 19 +/- 6 mm Hg at 14 mm Hg IAP, p < 0.005), but the transmural portal and hepatic venous pressures decreased (p < 0.01), indicating decreased venous filling. Portal flow was maintained at 7 mm Hg but decreased at 14 mm Hg from 474 +/- 199 to 395 +/- 175 mL/min (p < 0. 01), whereas hepatic arterial flow remained stable. Hepatic superficial blood flow decreased during insufflation and increased after desufflation. Tissue pH fell together with portal and hepatic venous pH (intestinal: from 7.323 +/- 0.05 to 7.217 +/- 0.04; hepatic: from 7.259 +/- 0.04 to 7.125 +/- 0.06, both p < 0.01) at 14 mm Hg. CONCLUSION: The hemodynamic and metabolic derangement in the pulmonary and splanchnic compartments are dependent on the extent of carbon dioxide pneumoperitoneum. The effect of low IAP (7 mm Hg) on splanchnic perfusion is minimal. However, higher IAPs (14 mm Hg) decrease portal and superficial hepatic blood flow and hepatic and intestinal tissue pH. PMID- 9869729 TI - Gastrointestinal ulceration and hemorrhage from hepatic arterial infusion chemotherapy. PMID- 9869730 TI - Endoscopic treatment of Zenker's diverticulum. AB - BACKGROUND: The most important aspect of the surgical management of Zenker's diverticulum is probably the cricopharyngeal myotomy. Endoscopic diverticulotomy can be performed with a needle-knife papillotome, which allows simultaneous myotomy of the upper esophageal sphincter. METHODS: Since 1978, 47 patients (28 men and 19 women 51 to 81 years of age) underwent endoscopic diverticulotomy. Most patients underwent more than one treatment session (mean value 2.2). The procedure was performed with sedation. Tubes were not used, and oral intake of food was begun the first day after the operation. RESULTS: Forty-five (95.74%) patients had no dysphagia or only occasional, mild dysphagia during the postoperative course. No fistula, no recurrent laryngeal paralysis, and no deaths occurred. CONCLUSION: Endoscopic diverticulotomy seems to be a good choice of therapy at least for patients with associated diseases that increase surgical risk. PMID- 9869731 TI - Endoscopic fluorescence detection of dysplasia in patients with Barrett's esophagus, ulcerative colitis, or adenomatous polyps after 5-aminolevulinic acid induced protoporphyrin IX sensitization. AB - BACKGROUND: Surveillance of patients with Barrett's esophagus or ulcerative colitis for dysplasia is confined to biopsy specimens taken randomly during endoscopy because dysplasia remains undetectable by visual inspection. We attempted to visualize dysplastic tissue during endoscopy after sensitization with 5-aminolevulinic acid (5-ALA) leading to accumulation and formation of protoporphyrin IX and induction of characteristic red fluorescence of the latter substance using blue light illumination. METHODS: Six patients with histologically proven low- or high-grade dysplasia (Barrett's esophagus 2, ulcerative colitis 1, Billroth-II stomach 1, rectal polyps 2) were treated with oral administration of different concentrations of 5-ALA (10 to 20 mg/kg) or by local instillation of 3 gm 5-ALA in the rectum. Endoscopic fluorescence detection was performed 1 to 6 hours after sensitization using a blue light source and compared with conventional white light endoscopy. Biopsies of fluorescent and nonfluorescent areas were compared with histologic findings. RESULTS: Normal duodenal mucosa and squamous epithelium showed more intense 5-ALA-induced background red fluorescence compared with normal mucosa in the stomach or Barrett's mucosa. Histologically, dysplasia was exclusively found in areas with red fluorescence. False-positive fluorescence was associated with microscopic inflammation of the mucosa or feces in the colon. CONCLUSIONS: 5-ALA-induced protoporphyrin IX fluorescence may be useful in the detection of dysplasia in the gastrointestinal tract by enhancement of endoscopic surveillance of patients at a high risk for dysplasia. PMID- 9869732 TI - Washout of small stones in the bile duct by saline infusion using a side-holed balloon catheter in patients undergoing endoscopic papillary balloon dilation. AB - BACKGROUND: Complete bile duct clearance of stones should be achieved in patients managed with endoscopic papillary balloon dilation. However, complete retrieval of small stones or tiny fragments sometimes proves difficult using conventional devices. METHODS: We attempted the removal of fine stone fragments by saline infusion using a specially designed retrieval balloon catheter with a blind tip and a side hole located proximal to the balloon in 14 patients. RESULTS: Using this technique, bile duct clearance was achieved in a single attempt in 13 of 14 patients; 2 attempts were required in 1 patient. Cholecystitis occurred in 2 patients with gallstones in situ after the procedure. CONCLUSIONS: Saline washout using a side-holed retrieval balloon catheter effectively cleans stones and stone fragments from the bile duct in patients treated with endoscopic papillary balloon dilation. PMID- 9869733 TI - Black esophagus associated with herpes esophagitis. PMID- 9869734 TI - Pericecal appendiceal abscess with drainage during colonoscopy. PMID- 9869735 TI - Occurrence of pyogenic meningitis during the course of endoscopic variceal ligation therapy. PMID- 9869736 TI - Endoscopic ligation by means of clip and detachable snare for management of colonic postpolypectomy hemorrhage. PMID- 9869737 TI - Treatment of gastric fundal varices by balloon endoscopic sclerotherapy. PMID- 9869738 TI - Lipomatosis of the small intestine and colon associated with intussusception in the ileocecal region. PMID- 9869739 TI - Ablation therapy for Barrett's esophagus: is it time to choose our weapons? PMID- 9869740 TI - Endoscopic treatment of Zenker's diverticulum. PMID- 9869741 TI - Introduction PMID- 9869743 TI - Comment PMID- 9869742 TI - Gastrointestinal endoscopy and the patient with a risk of bleeding disorder. PMID- 9869744 TI - Editor's comment PMID- 9869745 TI - Airway complication of expandable stents. PMID- 9869747 TI - Response PMID- 9869746 TI - Risk of self-expanding metal stents in patients with prior radiation or chemotherapy. PMID- 9869748 TI - Response PMID- 9869749 TI - Sclerotherapy in acute variceal bleeding. PMID- 9869750 TI - Masking of colon vascular ectasias by cold water lavage. PMID- 9869751 TI - Transgene expression after adenovirus-mediated retransfection of rat lungs is increased and prolonged by transplant immunosuppression. AB - OBJECTIVES: Adenovirus-mediated gene therapy has been proposed as a potential treatment modality in lung transplantation. However, to date its utility has been limited by an inflammatory host immune response that not only limits the amount and duration of transgene expression but also obviates successful retransfection. Having previously shown that by administering triple-immunosuppression, as is routine in lung transplantation, we could increase and prolong transgene expression after initial transfection, we hypothesized that transgene expression after retransfection could also be increased and prolonged. METHODS: Lewis rats underwent intratracheal adenovirus-mediated transfection with the beta galactosidase gene and were randomized to either the immunosuppression group, receiving daily cyclosporine (INN: ciclosporin), azathioprine, and methylprednisolone, or the control group (no immunosuppression). Five weeks later, rats were similarly retransfected and transgene expression and post transfection inflammation were evaluated 1, 7, and 14 days after retransfection. RESULTS: After retransfection, immunosuppressed rats had significantly higher levels of transgene expression (P <.001), whereas control rats had virtually no detectable levels. On histologic sections of the lungs, immunosuppressed rats had overall lesser grades of post-transfection inflammation. CONCLUSIONS: Transplant immunosuppression attenuates the severe immune response to gene transfer and permits increased, prolonged, and repeated transfection. Retransfection is now achievable in the immunosuppressed lung transplant setting to allow for chronic, repeated administration of gene therapy. PMID- 9869753 TI - Prevalence and location of nodal metastases in distal esophageal adenocarcinoma confined to the wall: implications for therapy. AB - OBJECTIVE: The purpose of this study was to characterize the prevalence and location of regional lymph node metastases in adenocarcinoma confined to the esophagal wall, to determine the extent of dissection required, and to investigate the applicability of nonoperative therapy. METHODS: Histologic evaluation of the resected specimens after en bloc esophagogastrectomy with mediastinal and abdominal lymphadenectomy was performed on 37 patients with adenocarcinoma confined to the esophageal wall. Follow-up was complete in all patients (median 24 months). RESULTS: Fifteen patients (41%) had intramucosal tumors. Twelve (32%) had submucosal tumors and 10 (27%) had muscular invasion. The prevalence of regional lymph node metastases (15/37 patients, 41%) increased progressively with depth of tumor invasion, with involved nodes identified in 80% of patients with muscular invasion. Lymph node metastases were also more common at distant node stations in intramuscular tumors (5/10, 50%). Actuarial survival for the entire group was 63% at 5 years. Recurrence was identified in 6 of the 37 patients (16%), with the risk of recurrence correlating with tumor depth. CONCLUSIONS: Tumor depth is a strong predictor of the probabilities of regional lymph node metastases, the likelihood of involvement of distant node groups, and the risk of recurrence. Patients with invasion of the muscular wall are at particularly high risk. En bloc esophagectomy with mediastinal and abdominal lymphadenectomy has the highest likelihood of achieving an R0 resection. The long term survival and low recurrence rate achieved with an en bloc esophagectomy emphasizes the importance of an aggressive lymph node dissection to remove all potentially involved nodes. PMID- 9869754 TI - Commentary: Esophageal carcinoma confined to the wall--the need for immediate definitive therapy. PMID- 9869752 TI - Liposome-mediated gene transfer in rat lung transplantation: A comparison between the in vivo and ex vivo approaches. AB - OBJECTIVE: We compared the efficacy of in vivo and ex vivo liposome transfection in rat lung transplantation. METHODS: (1) Chloramphenicol acetyltransferase group: Fischer rats underwent isogeneic transplantation (n = 4 per group). Recipients were put to death on postoperative day 2 for chloramphenicol acetyltransferase activity. Ex vivo setting: Grafts received cDNA complexed or not with liposomes and were transplanted after 1.5 or 10 hours at 10 degreesC. In vivo setting: Donors were intravenously injected with cDNA complexed or not with liposomes. Lungs were harvested after 1.5 or 10 hours, preserved at 10 degreesC, and transplanted. (2) Transforming growth factor-beta1 group: Brown-Norway rats served as donors and Fischer rats as recipients. All grafts were preserved for 3 hours at 10 degreesC. On postoperative day 5, arterial oxygenation and histologic rejection scores were assessed. Ex vivo setting: Grafts received transforming growth factor-beta1 sense (n = 8) or antisense (n = 7) complexed with liposomes or cDNA alone (n = 5). In vivo setting: Donors were intravenously injected with liposome:transforming growth factor-beta1 sense cDNA (n = 7). Exposure time was 3 hours. RESULTS: (1) Chloramphenicol acetyltransferase-transfection was superior in the ex vivo group but was not statistically different for longer exposure times. (2) Transforming growth factor-beta1-arterial oxygenation was superior in the ex vivo liposome:sense group. cDNA alone was inefficient. Rejection scores were not statistically different between ex vivo and in vivo liposome:sense groups but were better when the ex vivo liposome:sense group was compared with the cDNA alone or the antisense groups. CONCLUSIONS: (1) With current liposome technology, the ex vivo route is superior to the in vivo approach; (2) cDNA alone does not provide transgene expression at levels to produce a functional effect. PMID- 9869755 TI - Results of reoperation on the upper esophageal sphincter. AB - OBJECTIVE: Reoperation on the upper esophageal sphincter is infrequent. We reviewed our experience in patients who underwent reoperation on the upper esophageal sphincter. METHODS: This is a retrospective report of accumulative series from 2 separate institutions. RESULTS: From September 1, 1976, to February 28, 1997, 37 patients underwent reoperation on the upper esophageal sphincter for recurrent or persistent obstructive symptoms. There were 29 men and 8 women. The median age was 69 years (range, 38-87 years). The original indication for the operation was a pharyngoesophageal (Zenker's) diverticulum in 33 patients (89.2%), oculopharyngeal dystrophy in 3 patients (8.1%), and muscular dystrophy in 1 patient (2.7%). One prior upper esophageal sphincter operation had been performed in 26 patients (70.3%), two operations in 9 patients (24. 3%), and three operations in 2 patients (5.4%). All patients were symptomatic; 35 patients (94.6%) had dysphagia; 23 patients (62.2%) had regurgitation; and 12 patients (32.4%) had episodes of aspiration. Thirty of the patients (91.0%) with Zenker's diverticulum were found to have a recurrent or persistent diverticulum at reoperation. A diverticulectomy and cricopharyngeal myotomy were performed in 23 patients (62.2%); cricopharyngeal myotomy alone, in 7 patients (18.9%); diverticulopexy and cricopharyngeal myotomy, in 6 patients (16.2%); and diverticulectomy alone, in 1 patient (2.7%). There were no operative deaths. Complications developed in 10 patients (27.0%). Follow-up was complete in 34 patients (91.9%) and ranged from 2 to 149 months (median, 39 mo). Thirty-two patients (94.1%) were improved. Functional results were classified as excellent in 26 patients (76. 5%), good in 2 patients (5.9%), fair in 4 patients (11.7%), and poor in 2 patients (5.9%). CONCLUSIONS: Reoperation for patients who have persistent or recurrent symptoms after an operation on the upper esophageal sphincter is associated with acceptable morbidity and mortality rates. Resolution of symptoms occurs in most patients. PMID- 9869756 TI - Factors influencing ten-year survival in resected stages I to IIIa non-small cell lung cancer. AB - OBJECTIVE: The purpose of this study was to determine (in survivors of 5 years after resection of their lung cancer) whether age, sex, histologic condition, and age have any influence on furthering survival beyond 5 years. METHODS: From 1973 to 1989, 686 patients were alive and well 5 years after complete resection of their lung cancers. Survival analysis was carried out with only deaths from lung cancer treated as deaths. Deaths from other causes were treated as withdrawals. Multivariate Cox regression was used to test the relationship of survival to age, sex, histologic condition, and stage. RESULTS: The population in this study had the following characteristics at the time of operation: The male/female ratio was 1.38:1, and the median age was 61 years. The histologic condition of their lung cancer was adenocarcinoma in 412 patients, squamous cell in 244 patients, large cell carcinoma in 29 patients, and small cell carcinoma in 1 patient. The stage of the disease was stage IA in 263 patients, IB in 261 patients, IIA in 12 patients, IIB in 68 patients, and IIIA in 82 patients. The extent of resection was a lobectomy or bilobectomy in 579 patients, pneumonectomy in 55 patients, and wedge resection or segmentectomy in 52 patients. A recurrence or a new lung primary occurrence was considered as failure to remain free of lung cancer. The median follow-up on all patients was 122 months from initial treatment. Of the 686 patients, 26 patients experienced the development of late recurrence and 36 new cancers, beyond 5 years. Overall survival for 5 additional years after a 5 year check point was 92.4%. Likewise, survival by nodal status was 93% for N0 tumors, 95% for N1 tumors, and 90% for N2 tumors. Survival by stage was 93% for stage I tumors and 91% for stage II or IIIA tumors. CONCLUSIONS: In patients with surgically treated lung cancer, neither age, sex, histologic condition, nor stage is a predictor of the risk of late recurrence or new lung cancer. The only prognostic factor appears to be the survival of the patient free of lung cancer for 5 years from the initial treatment, with a resultant favorable outlook to remain well for 10 or more years. PMID- 9869757 TI - Clinical experience with carinal resection. AB - OBJECTIVE: Pathologic processes that involve the carina pose a tremendous challenge to thoracic surgeons. Although techniques have been developed to allow primary resection and reconstruction, few institutions have accumulated sufficient experience to allow meaningful conclusions about the indications and the morbidity and mortality rates for this type of surgery. METHODS: Since 1962, 135 patients have undergone 143 carinal resections (134 primary resection, 9 re resection) at our institution. Indications for carinal resection included bronchogenic cancer (58 patients), other airway neoplasms (60 patients), and benign or inflammatory strictures (16 patients). Thirty-seven patients (28%) had a history of prior lung or airway surgery not involving the carina. Carinal resection without pulmonary resection was accomplished in 52 patients; 57 patients had carinal pneumonectomy (44 right, 13 left); 14 patients had carinal plus lobar resection, and 11 patients had carinal resection after pneumonectomy (9 left, 2 right). There were 15 different modes of reconstruction, based on the type and extent of resection. Techniques were used to reduce anastomotic tension. RESULTS: The operative mortality rate in the 134 patients after primary carinal resection was 12.7%. Adult respiratory distress syndrome was responsible for 9 early deaths. Predominant predictors of operative death included postoperative mechanical ventilation (P =.001), length of resected airway (P =.03), and development of anastomotic complications (P =.04). Mortality rates varied by the type of procedure and the indication for resection. Left carinal pneumonectomy was associated with a high operative mortality rate (31%). Complications were noted in 52 patients (39%), including atrial arrhythmias (20 patients) and pneumonia (11 patients). Anastomotic complications, both early and late, were seen in a total of 23 patients (17%) and resulted in death or surgical reintervention in 21 patients (91%). The operative mortality rate for carinal re resection was 11.1%. CONCLUSIONS: Carinal resection with primary reconstruction may be accomplished with acceptable mortality rates, but the underlying pathologic process and chance for long-term survival must be carefully considered before the operation is recommended, especially in the case of left carinal pneumonectomy. Anastomotic complications exact a heavy toll on involved patients. Careful patient selection and meticulous anesthetic and surgical technique remain the key to minimizing morbidity and mortality rates. PMID- 9869758 TI - Resection margins, extrapleural nodal status, and cell type determine postoperative long-term survival in trimodality therapy of malignant pleural mesothelioma: results in 183 patients. AB - OBJECTIVES: Our aim was to identify prognostic variables for long-term postoperative survival in trimodality management of malignant pleural mesothelioma. METHODS: From 1980 to 1997, 183 patients underwent extrapleural pneumonectomy followed by adjuvant chemotherapy and radiotherapy. RESULTS: Forty three women and 140 men (age range 31-76 years) had a median follow-up of 13 months. The perioperative mortality rate was 3.8% (7 deaths) and the morbidity, 50%. Survival in the 176 remaining patients was 38% at 2 years and 15% at 5 years (median 19 months). Univariate analysis identified 3 prognostic variables associated with improved survival: epithelial cell type (52% 2-year survival, 21% 5-year survival, 26-month median survival; P =.0001), negative resection margins (44% at 2 years, 25% at 5 years, median 23 months; P =.02), and extrapleural nodes without metastases (42% at 2 years, 17% at 5 years, median 21 months; P =.004). Using the Cox proportional hazards, the relative risk of death was calculated for nonepithelial cell type (OR 3.0, CI 2.0-4.5; P <.0001), positive resection margins (OR 1.7, CI 1.2-2.6; P =.0082), and metastatic extrapleural nodes (OR 2.0, CI 1.3-3.2; P =.0026). Thirty-one patients with 3 positive variables had the best survival (68% 2-year survival, 46% 5-year survival, median 51 months; P =.013). A previously published staging system using these variables stratified survival (P <.05). CONCLUSIONS: (1) Multimodality therapy including extrapleural pneumonectomy is feasible in selected patients with malignant pleural mesotheliomas, (2) pre-resectional evaluation of extrapleural nodes may select patients for radical therapy, (3) microscopic resection margins affect long-term survival, highlighting the need for further investigation of locoregional control, and (4) patients with epithelial, margin-negative, extrapleural node-negative resection had extended survival. PMID- 9869759 TI - Is resection of pulmonary and hepatic metastases warranted in patients with colorectal cancer? AB - BACKGROUND: Conventional management of stage IV colorectal carcinoma is palliative. The value of resecting both liver and lung colorectal metastases that occur in isolation of other sites of metastasis is undetermined. OBJECTIVES: Our objectives were to (1) assess the efficacy of resecting both hepatic and pulmonary metastases, (2) investigate the influence of the sequence and timing of metastases, and (3) identify the profile of patients likely to benefit from both hepatic and pulmonary metastasectomy. PATIENTS AND METHODS: Of 48 patients identified with resection of colorectal cancer and, at some point in time, both liver and lung metastases, 25 patients underwent metastasectomy (resection group). The remaining 23 patients comprised the nonresection group. Risk factors for death were identified by multivariable analyses. RESULTS: Median survival was longer after the last metastatic appearance in the resection group (16 months) than in the nonresection group (6 months; P <.001). The pattern of risk also differed; it peaked at 2 years and then declined in the resection group but was constant in the nonresection group. In the resection group, patients with metachronous resections survived longer after colorectal resection (median, 70 months) than patients with synchronous (median, 22 months) or mixed resections (median, 31 months; P <.001). Risk factors for death included older age, multiple liver metastases, and a short disease-free interval. CONCLUSIONS: Younger patients with solitary metachronous metastases to the liver, then the lung, and long disease-free intervals are more likely to benefit from resection of both liver and lung metastases. Patients with risk factors also had better survival with resection than without resection. PMID- 9869760 TI - Primary aortic valve replacement with allografts over twenty-five years: valve related and procedure-related determinants of outcome. AB - OBJECTIVES: Allografts offer many advantages over prosthetic valves, but allograft durability varies considerably. METHODS: From 1969 through 1993, 618 patients aged 15 to 84 years underwent their first aortic valve replacement with an aortic allograft. Concomitant surgery included aortic root tailoring (n = 58), replacement or tailoring of the ascending aorta (n = 56), and coronary artery bypass grafting (n = 87). Allograft implantation was done by means of a "freehand" subcoronary technique (n = 551) or total root replacement (n = 67). The allografts were antibiotic sterilized (n = 479), cryopreserved (n = 12), or viable (unprocessed, harvested from brain-dead multiorgan donors or heart transplant recipients, n = 127). Maximum follow-up was 27.1 years. RESULTS: Thirty-day mortality was 5.0%, and crude survival was 67% and 35% at 10 and 20 years. Ten- and 20-year rates of freedom from complications were as follows: endocarditis, 93% and 89%; primary tissue failure, 62% and 18%; and redo aortic valve replacement, 81% and 35%. Multivariable Cox analyses identified several valve- and procedure-related determinants: rising allograft donor age and antibiotic-sterilized allograft for mortality; donor more than 10 years older than patient for endocarditis; rising donor age minus patient age, rising implantation time (from harvest to aortic valve replacement), and donor age more than 65 years for tissue failure; and rising donor age minus patient age, young patient age, rising implantation time, and subcoronary implantation preceded by aortic root tailoring for redo aortic valve replacement. Estimated 10- and 20 year rates of freedom from tissue failure for a 70-year-old patient with a viable valve from a 30-year-old donor and no other risk factors were 91% and 64%; the figures were 71% and 20% if the donor age was 65 years. The rates of freedom from tissue failure for a 30-year-old patient with a 30-year-old donor were 82% and 39%; the figures were 49% and 3% with a 65-year-old donor. Beneficial influences of a viable valve were largely covered by short harvest time (no delay for allografts from brain dead organ donors or heart transplant recipients) and short implantation time. CONCLUSIONS: Primary allograft aortic valve replacement can give acceptable results for up to 25 years. The late results can be improved by the use of a viable allograft, by matching patient and donor age, and by more liberal use of free root replacement with re-implantation of the coronary arteries rather than tailoring the root to accommodate a subcoronary implantation. PMID- 9869761 TI - The effect of sizing on the in vitro hydrodynamic characteristics and leaflet motion of the Toronto SPV stentless valve. AB - OBJECTIVES: We established an in vitro model to investigate the effects of valve sizing on the hemodynamic characteristics and leaflet motion of the Toronto SPV valve (St Jude Medical, Inc, St Paul, Minn). METHODS: Nine valves were first implanted in fresh porcine aortic roots and then retested in glutaraldehyde treated porcine aortic roots. Three valves were 1- to 2-mm oversized, 3 were 1- to 2-mm undersized, and there were 3 size-for-size implantations. The elasticities of the aortic roots and the composite roots were measured in the pressure range between 0 and 120 mm Hg, and the composite roots were then tested in a pulsatile flow simulator. The transvalvular gradient and regurgitation were measured and the effective orifice area and performance index were calculated for each root. Leaflet motion was recorded on videotape. RESULTS: The external diameter of the fresh root increased by 35% as the hydrostatic pressure rose from 0 to 120 mm Hg, as compared with 11% for the glutaraldehyde-treated root. Valve implantation in the fresh root reduced the distensibility to 22% but did not change distensibility in the glutaraldehyde-treated root. The effective orifice area was dependent on the valve size, with the transvalvular gradient decreasing as the valve size increased. For the same size of valve the hydrodynamic parameters were slightly better if the valve was undersized by 1 mm. A significant difference in favor of the undersized valves was found in open leaflet bending deformation. CONCLUSION: Leaflet motion of the stentless porcine aortic valve in vitro is improved if the valve is slightly undersized, and this may be beneficial to the long-term durability of the prosthesis. PMID- 9869762 TI - Single-stage extensive replacement of the thoracic aorta: the arch-first technique. AB - BACKGROUND: Single-stage extensive replacement of the thoracic aorta usually involves a period of circulatory arrest with performance of the graft-to-lower descending thoracic aorta anastomosis before performing the anastomosis to the arch vessels. To minimize the period of brain ischemia and reduce the potential for neurologic injury, we developed an alternative technique. METHODS: In 6 patients with extensive aneurysms involving the entire thoracic aorta, exposure was obtained via a bilateral thoracotomy in the anterior fourth intercostal space with transverse sternotomy. A 10-mm graft was anastomosed to the aortic graft, opposite the site of the planned anastomosis to the arch vessels. During a single period of circulatory arrest (34-46 minutes), the aortic graft was attached to a cuff of aorta containing the arch vessels. The graft was then clamped on either side, and the arch was perfused with cold blood for 20 to 36 minutes. After the distal aortic anastomosis was completed, antegrade perfusion was established via the 10-mm graft. The proximal aortic anastomosis was performed last. RESULTS: No patient sustained a permanent neurologic deficit. All 6 patients were discharged from the hospital. CONCLUSIONS: The "arch-first" technique, combined with a bilateral transverse thoracotomy, allows expeditious replacement of the thoracic aorta with an acceptable interval of hypothermic circulatory arrest and minimizes the risk of retrograde atheroembolism by establishing antegrade perfusion. PMID- 9869763 TI - Cardiac valve papillary fibroelastoma: surgical excision for revealed or potential embolization. AB - OBJECTIVE: We have reviewed the case histories of 4 patients who underwent operations between September 1994 and November 1997 at Broussais Hospital for cardiac valvular papillary fibroelastoma. METHODS: Diagnosis was strongly suggested by echocardiography. Tumor locations were mitral (1), tricuspid (1), and aortic (2). Indications for operation were previous stroke for the mitral tumor, prophylaxis for the tricuspid tumor, syncopal episodes for the first aortic tumor, and transient ischemic attack and mesenteric ischemia for the second aortic tumor. RESULTS: Surgical excision with a conservative, valve sparing approach was performed in all cases. For the first aortic tumor, aortic valve reconstruction was achieved with part of a cryopreserved aortic homograft cusp. Intraoperative transesophageal echocardiography showed no evidence of valvular regurgitation after excision in all cases. All patients had uneventful postoperative recoveries. No evidence of regurgitation or recurrence was seen on echocardiography at follow-up. CONCLUSIONS: Despite their histologically benign aspect, cardiac papillary fibroelastomas should be excised because of potential embolic complications. A conservative, valve-sparing approach is recommended, however, because of the absence of recurrence after total excision. PMID- 9869764 TI - Atherosclerosis of the ascending aorta is a predictor of renal dysfunction after cardiac operations. AB - OBJECTIVES: Renal dysfunction occurring after cardiac operations has been attributed to various factors, but the importance of an atherosclerotic thoracic aorta has not been previously evaluated. The purpose of this study was to identify predictors of postoperative renal dysfunction (50% or more increase from preoperative values) and to evaluate the importance of atherosclerosis of the ascending aorta as a predictor of this complication. METHODS: Nine hundred seventy-eight consecutive patients, 50 years of age and older with normal preoperative renal function (serum creatinine level of 1.5 mg/dL or less), who were scheduled to undergo cardiac surgery were prospectively evaluated. Atherosclerosis of the ascending aorta was assessed during the operation (with epiaortic ultrasound), and patients were divided into 3 groups according to its severity (normal-to-mild, moderate, and severe). RESULTS: Univariate predictors of renal dysfunction at postoperative day 1 were atherosclerosis of the ascending aorta (P <. 045) and postoperative low cardiac output (P =.05); at postoperative day 6 they were atherosclerosis of the ascending aorta (P <.0001), postoperative low cardiac output (P <.0001), advanced age (P =.001), decreased preoperative left ventricular function (P =.01), and female gender (P =.03). Multivariate analysis showed that atherosclerosis of the ascending aorta (odds ratio, 3.06; P =.04) was the only independent predictor of postoperative renal dysfunction at day 1 and that postoperative low cardiac output (odds ratio, 4.83; P <.0001), atherosclerosis of the ascending aorta (odds ratio, 2.13; P =.0006), and preoperative left ventricular dysfunction (odds ratio, 1.48; P =.028) were independent predictors of postoperative renal dysfunction at day 6. CONCLUSIONS: An atherosclerotic ascending aorta is an important predictor of postoperative renal dysfunction, possibly because atheroembolism to the kidneys occurs in the perioperative period (ie, during surgical manipulation of an atherosclerotic aorta) or because the diseased aorta may be a marker of widespread atherosclerotic disease that may predispose to perioperative renal dysfunction. PMID- 9869765 TI - A novel one-shot anastomotic stapler prototype for coronary bypass grafting on the beating heart: feasibility in the pig. AB - OBJECTIVE: The nonpenetrating, arcuate-legged clip has proved its ability to provide a high-quality microvascular anastomosis. This study assessed the feasibility of constructing a coronary end-to-side anastomosis on the beating heart with a novel mechanical, sutureless anastomotic device that applies 12 circumferential clips simultaneously. METHODS: In 14 consecutive pigs (70-90 kg), the left internal thoracic artery (diameter, 3 mm) was grafted to the left anterior descending coronary artery (diameter, 3 mm) by means of a one-shot anastomotic stapler prototype. Endothelial denudation, medial necrosis, and intimal hyperplasia were analyzed quantitatively and compared with those seen in conventionally sutured anastomoses (n = 4). RESULTS: In 8 of 14 anastomoses, the one-shot anastomotic stapler successfully applied all 12 clips circumferentially across the everted arteriotomy edges. In the remaining, either 1 (n = 4) or 3 and 4 adjoining malaligned clips had to be replaced manually with a single-clip applicator. Coronary occlusion was limited to approximately 3 minutes. At follow up, all anastomoses were patent angiographically. At 2 days, in 2 of 7 cases, a local coronary dissection was observed, and there was a considerable loss of endothelial cells and medial damage. At 28 days, however, minimal intimal hyperplasia was seen at the anastomotic lining, although more pronounced when compared with conventionally sutured anastomoses. CONCLUSIONS: The one-shot anastomotic stapler prototype enabled short-occlusive (3 minutes), sutureless end to-side grafting on the beating porcine heart. In spite of early endothelial and medial damage and 2 local dissections, all anastomoses remained patent with minimal intimal hyperplasia at 4 weeks. PMID- 9869766 TI - Mobilization of the left and right fibrous trigones for relief of severe left ventricular outflow obstruction. AB - BACKGROUND: There is still no agreement about the optimal method of surgical relief of fixed subaortic stenosis, particularly the severe forms. OBJECTIVES: The purpose of this study was to describe a new technique for the relief of subaortic stenosis based on analysis of the functional anatomy of the left ventricular outflow tract and pathophysiologic features of subaortic stenosis. METHODS AND PATIENTS: We propose that one of the basic abnormalities in subaortic stenosis is interference with the hinge mechanism provided by the 2 fibrous trigones with progressive deposition of fibrous tissue in these angles. The technique described in this paper consists of excision of all components of the fibrous "ring," with mobilization of the left and right fibrous trigones. This results in the restoration of the normal dynamic behavior of the left ventricular outflow tract with maximal widening of the outflow tract as the result of backward displacement of the subaortic curtain and anterior leaflet of the mitral valve. This technique has been used in 57 consecutive patients who ranged in age between 5 months and 56 years (mean, 15.5 +/- 10.6 years). Gradients across the left ventricular outflow tract were between 45 and 200 mm Hg (mean, 86.7 mm Hg). Additional lesions were present in 10 patients, and 7 patients had had 8 previous operations on the left ventricular outflow tract. At operation, in addition to resection of subaortic stenosis, 3 patients had aortic valvotomy, 2 patients had homograft replacement of the aortic valve, 7 patients had patch closure of a ventricular septal defect, and 1 patient had open mitral valvotomy. RESULTS: There were 2 early deaths and 1 late sudden death during the follow-up period that ranged from 1 month to 25 years (mean, 15. 2 years). One patient experienced the development of endocarditis on the aortic valve 7 years after operation, which was successfully treated by homograft replacement. Postoperative gradients across the left ventricular outflow tract varied from no gradient to 30 mm Hg (mean, 8 mm Hg). There were no instances of recurrence of a gradient across the left ventricular outflow tract. CONCLUSION: It is concluded that mobilization of the left and right fibrous trigones results in durable relief of subaortic stenosis. PMID- 9869767 TI - Multiple ventricular septal defects: how and when should they be repaired? AB - BACKGROUND: Congenital heart lesions with multiple ventricular septal defects remain a surgical challenge. Traditional approaches often rely on either ventriculotomy for exposure or palliation with pulmonary artery banding. However, indications for repair versus palliation and for various approaches to surgical exposure are not clearly defined. METHODS: From July 1992 to January 1998, 45 patients with multiple (>/=2) ventricular septal defects (37 with associated lesions) underwent surgery. Median age was 86 days; all but 4 patients were infants. The mean number of defects was 3.7, and almost half of the patients had more than 3 defects. Apical muscular defects were present in 62% of patients. Thirty-one patients underwent primary complete repair through a right atriotomy or trans-semilunar valve approach (group 1), 8 had palliation (group 2), and 6 underwent complete repair after prior palliation elsewhere (group 3). No patient had a ventriculotomy. RESULTS: One early death occurred in a group 1 patient. Four patients who had had palliation (50%) underwent early reoperation for pulmonary artery band revision because of failure to thrive or band removal after spontaneous closure of the defects. At follow-up (median 22 months), there was 1 death in a group 2 patient (palliation) and 1 other group 2 patient required cardiac transplantation. The only late reoperation was for removal of the pulmonary artery band and closure of multiple apical defects in a group 2 (palliation) patient. No patients who underwent repair have hemodynamically significant residual defects. CONCLUSIONS: In our experience, palliation of multiple ventricular septal defects is associated with greater morbidity than primary repair. Multiple defects can almost always be repaired adequately in early infancy without ventriculotomy, although "Swiss-cheese" septum may be an indication for palliation. PMID- 9869768 TI - Cryopreserved homograft valves in the pulmonary position: risk analysis for intermediate-term failure. AB - OBJECTIVE: The purpose of this study was to examine the durability of cryopreserved homografts used to replace the "pulmonary" valve and to identify factors associated with their late deterioration. METHODS: We reviewed our entire experience (1985-1997) with 331 survivors in whom cryopreserved homograft valves (pulmonary, n = 304; aortic, n = 27) were used to reconstruct the pulmonary outflow tract. Median age was 14 years (range, 2 days-62 years). Operations included Ross operation (n = 259), tetralogy of Fallot (n = 41), truncus arteriosus (n = 14), Rastelli operation (n = 11), and others (n = 6). Median follow-up was 3.8 years (range, 0.2-11.2 years); late echographic follow-up was complete for 97% of patients. Homograft failure was defined as the need for explantation and valve-related death; homograft dysfunction was defined as a pulmonary insufficiency grade 3/4 or greater and a transvalvular gradient of 40 mm Hg or greater. RESULTS: Homograft failure occurred in 9% (30 of 331 patients; Kaplan-Meier); freedom from failure was 82% +/- 4% at 8 years. Homograft dysfunction occurred in 12% (39 of 331 patients), although freedom from dysfunction was 76% +/- 4% at 8 years. For aortic homografts, this was 56% +/- 11%, compared to 80% +/- 4% for pulmonary homografts (P =.003). For patients aged less than 3 years (n = 38), this was 51% +/- 12%, compared with 87% +/- 4% for older patients (P =.0001). By multivariable analysis, younger age of homograft donors, non-Ross operation, and later year of operation were associated with homograft failure; younger age of homograft donors, later year of operation, and use of an aortic homograft were associated with homograft dysfunction. CONCLUSIONS: Homograft valves function satisfactorily in the pulmonary position at mid-term follow-up. The pulmonary homograft valve appears to be more durable than the aortic homograft valve in the pulmonary position. PMID- 9869769 TI - Morphologic spectrum of Ebstein's malformation: revisitation relative to surgical repair. AB - OBJECTIVE: Our aim was to elucidate the morphologic spectrum of Ebstein's malformation of the tricuspid valve with regard to diagnosis and the feasibility of surgical repair, in the light of the currently favored reconstructive techniques. METHODS: We examined 23 autopsied hearts. Taking the displacement of the septal and mural leaflets of the abnormal tricuspid valve as our diagnostic criterion, we focused subsequently on the location of the distorted valvular orifice and the attachment and formation of the anterosuperior leaflet. We also assessed the dimensions of the components of the right ventricle relative to the plane of the displaced valvular orifice. RESULTS: In all hearts, the septal and mural leaflets were hinged at various points within the inlet of the right ventricle. In many cases, however, these leaflets were virtually absent. The plane of the effective tricuspid valvular orifice was displaced anterosuperiorly to varying degrees. In the most severe forms, the valvular mechanism took the form of a 1-leaflet valve. The length of the functional right ventricle when compared with the left ventricle ranged proportionally from 0.6 to 1. 1 (mean, 0.9). CONCLUSIONS: Ebstein's malformation is much more than simple "downward displacement" of the leaflets. In essence, the valvular orifice is formed within the ventricular cavity at the junction of the atrialized inlet and functional ventricular components. When surgical intervention becomes necessary, it is essential to make a detailed assessment of both valvular and ventricular abnormalities. PMID- 9869770 TI - Neuropsychologic outcome after deep hypothermic circulatory arrest in adults. AB - INTRODUCTION: Pediatric patients undergoing prolonged periods of deep hypothermic circulatory arrest have been found to experience long-term deficits in cognitive function. However, there is limited information of this type in adult patients who are undergoing deep hypothermic circulatory arrest for thoracic aortic repairs. METHODS: One hundred forty-nine patients undergoing elective cardiac or thoracic aortic operations were evaluated preoperatively; 106 patients were evaluated early in the postoperative period (EARLY), and 77 patients were evaluated late in the postoperative period (LATE) with a battery of neuropsychologic tests. Seventy-three patients had routine cardiac operations without deep hypothermic circulatory arrest, and 76 patients with deep hypothermic circulatory arrest were divided into 2 subgroups: those with 1 to 24 minutes of deep hypothermic circulatory arrest (n = 36 patients) and those with 25 minutes or more of deep hypothermic circulatory arrest (n = 40 patients). The neuropsychologic test battery consisted of 8 tests encompassing 5 domains: attention, processing speed, memory, executive function, and fine motor function. Data were normalized to baseline values, and changes from baseline were analyzed by analysis of covariance, multivariate logistic regression, and survival functions. RESULTS: In all domains, poor performance or inability to be tested EARLY were significant predictors of poor performance LATE (odds ratio, 5.27; P <.01). Deep hypothermic circulatory arrest of 25 minutes or more and advanced age were significant predictors of poor performance LATE for the memory and fine motor domains. Deep hypothermic circulatory arrest of 25 minutes or more (odds ratio, 4. 0; P =.02) was a determinant of prolonged hospital stay (>21 days). CONCLUSION: Deep hypothermic circulatory arrest of 25 minutes or more and advanced age were associated with memory and fine motor deficits and with prolonged hospital stay. PMID- 9869771 TI - Ventriculoarterial coupling with intra-aortic balloon pump in acute ischemic heart failure. AB - PURPOSE: We analyzed the mechanism of effects of intra-aortic balloon pumping using the pressure-volume relationship and ventriculoarterial coupling in the normal and failing hearts. MATERIALS: In 12 anesthetized Holstein calves (weight, 94 +/- 8 kg), the ventricular end-systolic and arterial elastances, pressure volume area, and external work were analyzed during steady-state contractions with traditional hemodynamic parameters with intra-aortic balloon pumping-off and -on (1:1 synchronous ratio). An acute ischemic heart failure was induced by injecting 10 microm microspheres (4.2 +/- 1.8 x 10(7). 100g left ventricular weight-1) into the left main coronary artery; all measurements were repeated. RESULTS: Intra-aortic balloon pumping did not change hemodynamic parameters in the control. However, during heart failure, intra-aortic balloon pumping decreased the arterial elastance from 3.6 +/- 1.3 mm Hg to 2.9 +/- 1.2 mm Hg. mL 1 while not affecting the ventricular end-systolic elastance, this resulted in an improvement of the ventriculoarterial coupling ratio from 3.1 +/- 0.8 to 2.3 +/- 0.8. Intra-aortic balloon pumping decreased not only end-systolic pressure (from 69 +/- 16 mm Hg to 64 +/- 19 mm Hg) but end-diastolic volume and pressure (from 139 +/- 38 mL to 137 +/- 37 mL and from 13. 9 mm Hg to 12.8 mm Hg, respectively) with the leftward shift of the pressure-volume loop. Pressure-volume area decreased (from 914 +/- 284 mm Hg to 849 +/- 278 mm Hg. mL) although stroke volume increased (from 21 +/- 6 mL to 24 +/- 6 mL). CONCLUSION: Reduction of the arterial elastance with intra-aortic balloon pumping improved the ventriculoarterial coupling ratio and increased stroke volume. Leftward shift of the pressure-volume loop resulted in the reduction of pressure-volume area, which suggests the conservation of the myocardial oxygen consumption. PMID- 9869773 TI - Tolerance of a resorbable collagen-elastin membrane as a pericardial substitute in adult cardiac operations. PMID- 9869772 TI - Comparison of the effects of transfusions of cryopreserved and liquid-preserved platelets on hemostasis and blood loss after cardiopulmonary bypass. AB - OBJECTIVE: The aim of the study was to compare the clinical effects and hemostatic efficiency of transfusions of platelets preserved in the frozen state for as long as 2 years with transfusions of platelets preserved in the conventional manner for as long as 5 days in patients undergoing cardiopulmonary bypass. METHODS: Seventy-three patients were prospectively randomly assigned to receive transfusions of cryopreserved or liquid-preserved platelets. Nonsurgical blood loss was measured during and after the operation. Bleeding time, hematologic variables, and the bleeding time site shed blood were assayed before cardiopulmonary bypass and at 30 minutes and 2, 4, and 24 hours after transfusion. In vitro platelet function tests were conducted on platelets obtained from healthy volunteers. RESULTS: No adverse sequelae of the transfusions were observed. Blood loss and the need for postoperative blood product transfusions were lower in the group receiving cryopreserved platelets. Lower posttransfusion platelet increments and a tendency toward decreased platelet survival were observed in patients receiving cryopreserved platelets. Hematocrit and plasma fibrinogen were significantly higher in this group, and the duration of intubation was shorter. In vitro, cryopreserved platelets demonstrated less aggregation, lower pH, and decreased response to hypotonic stress but generated more procoagulant activity and thromboxane. CONCLUSIONS: (1) Cryopreserved platelet transfusions are superior to liquid-preserved platelets in reducing blood loss and the need for blood product transfusions after cardiopulmonary bypass. (2) The reduction in blood loss in the patients receiving cryopreserved platelet transfusions after cardiopulmonary bypass probably reflects improved in vivo hemostatic function of cryopreserved platelets. (3) Some in vitro measures of platelet quality (aggregation, pH, hypotonic stress) may not reflect in vivo quality of platelet transfusions after cardiopulmonary bypass, whereas other in vitro measures (platelet procoagulant activity and thromboxane) do. PMID- 9869774 TI - Transapical aortic occlusion for cardioplegic delivery during reconstruction of thoracoabdominal aortic aneurysm with deep hypothermic circulatory arrest. PMID- 9869775 TI - Modified hemi-Fontan procedure on the beating heart. PMID- 9869776 TI - In situ implantation of a prosthetic systemic atrioventricular valve in corrected transposition. PMID- 9869777 TI - Tetralogy of Fallot with absent pulmonary valve: a surgical technique for complete repair. PMID- 9869778 TI - Effect of inhaled nitric oxide on left ventricular and pulmonary vascular function. PMID- 9869779 TI - Deep hypothermic circulatory arrest and retrograde cerebral perfusion. PMID- 9869780 TI - Reconstruction of the aortic valve with autologous pericardium: an experimental study. PMID- 9869781 TI - A stable model of chronic bilateral ventricular insufficiency (dilated cardiomyopathy) induced by arteriovenous anastomosis and doxorubicin administration in sheep. PMID- 9869782 TI - High hematocrit value is a risk factor for myocardial infarction. PMID- 9869783 TI - Anterior segment implantation cysts. Ultrasound biomicroscopy with histopathologic correlation. AB - OBJECTIVE: To correlate the clinical, histopathologic, and ultrasound biomicroscopic characteristics of anterior segment implantation cysts. METHODS: We performed a retrospective review of 7 cases of secondary anterior segment implantation cysts. We reviewed the clinical history, visual acuity, clinical findings, and ultrasound biomicroscopic characteristics in all cases. Histopathologic correlation was possible in 4 cases. RESULTS: Six eyes had been subjected to major trauma prior to cyst formation. Trauma was noted as blunt in 3 eyes and surgical in 3 eyes. The diagnosis was confirmed in 1 eye when conjunctival cells were aspirated on fine needle biopsy. Ultrasound biomicroscopy revealed large (mean +/- SD greatest diameter, 4.7 +/- 0.9 mm) cystic tumors. In 1 patient, a cyst-related indentation of the anterior lens surface was seen. Ultrasonographic evaluations of internal reflectivity revealed thick, moderately reflective cyst walls encapsulating a relatively hypoechoic core. In 3 cases, the cyst contents consisted of variably reflective material. The other 4 were completely sonolucent. Histopathologic correlation showed that the cyst walls were lined with stratified squamous epithelium. The moderately reflective cyst contents were found to be degenerated conjunctival cells with inflammatory foci and cholesterol crystals. The sonolucent regions correlated with inflammatory cells and fluid. CONCLUSIONS: This study demonstrates that implantation cysts are unilateral, large, and thick walled. They may be sonolucent or exhibit variable internal reflectivity. These findings as well as the extent of anterior segment involvement (particularly posterior extension) could be evaluated by ultrasound biomicroscopy prior to surgery. PMID- 9869784 TI - Quantitative comparison of posterior capsule opacification after polymethylmethacrylate, silicone, and soft acrylic intraocular lens implantation. AB - OBJECTIVE: To quantitatively compare the extent of posterior capsule opacification (PCO) after polymethylmethacrylate (PMMA), silicone, and soft acrylic intraocular lens implantation. PATIENTS AND METHODS: A total of 240 eyes from 240 patients undergoing implant surgery were randomized into 3 groups based on the type of lens implanted: PMMA, silicone, and soft acrylic. The density value of PCO in 185 eyes was quantitated approximately 2 years after surgery by a new measurement method using the Scheimpflug videophotography system. RESULTS: Twenty-one eyes (30.4%) in the PMMA group, 4 (5.7%) in the silicone group, and 2 (2.7%) in the acrylic group had already undergone Nd:YAG laser posterior capsulotomy. The mean +/- SD PCO values were 26.3 +/- 12.2 computer-compatible tape steps (CCT) in the PMMA group, 12.0 +/- 8.3 CCT in the silicone group, and 16.0 +/- 10.3 CCT in the acrylic group. The PCO value in the PMMA group was significantly greater than that in the silicone or acrylic group (P < .001). The visual acuity loss in the PMMA group was also greater than that in the silicone or acrylic group (P < .001). CONCLUSION: Based on the PCO value and capsulotomy rate, the PCO was more extensive with the PMMA lens than with either the silicone or soft acrylic lens, which led to visual acuity loss. PMID- 9869785 TI - Detection of glaucoma with scanning laser polarimetry. AB - OBJECTIVE: To determine which retinal nerve fiber layer (RNFL) measures obtained with scanning laser polarimetry are most useful in detecting early to moderate glaucomatous visual field loss. SUBJECTS AND METHODS: One eye from 84 healthy individuals and 83 patients with early to moderate glaucomatous visual field loss (167 eyes) was assessed with a scanning laser polarimeter (Laser Diagnostic Technologies, San Diego, Calif). Three separate scans were obtained, and a baseline scan was created and used in the analyses. Integrated software (program GDx, version 1.0.02; Laser Diagnostic Technologies) was evaluated by assessing its sensitivity and specificity for detecting early and moderate glaucomatous visual field loss. Fisher linear discriminant functions also were developed in this population to assess sensitivity and specificity and were compared with the GDx analyses. RESULTS: There were statistically significant differences between the healthy and glaucomatous eyes for 14 of the 15 RNFL measures (P = .001). However, considerable overlap in measurements between groups was found. With the GDx number, the area under the receiver operator characteristic (ROC) curve was 0.78, and the sensitivity and specificity were 82% and 62%, respectively. Applying the best discriminant function using 3 variables (average thickness, ellipse modulation, and average ellipse thickness) to our study population resulted in an area under the ROC curve of 0.89 and a sensitivity and specificity of 74% and 92%, respectively. CONCLUSIONS: A combination of RNFL measures obtained using the scanning laser polarimeter improved the ability to differentiate between healthy eyes and eyes with early and moderate glaucomatous visual field loss. Analyses using GDx software did not differentiate between healthy and glaucomatous eyes as well as the discriminant analysis function did. PMID- 9869787 TI - Interferon-induced protein 10 and interleukin 8. C-X-C chemokines present in proliferative diabetic retinopathy. AB - OBJECTIVE: To determine vitreous levels of interleukin 8 (IL-8) and interferon induced protein 10 (IP-10), which are members of the C-X-C chemokine family that promote and inhibit neovascularization, respectively. METHODS: We measured the levels of IL-8 and IP-10 by specific enzyme-linked immunosorbent assays in the vitreous from 30 patients with proliferative diabetic retinopathy (PDR) and 10 control patients undergoing vitrectomy for idiopathic macular holes or idiopathic macular puckers. RESULTS: Detectable levels of IL-8 were found in 23 of 24 patients with active PDR, 4 of 6 patients with inactive PDR, and 6 of 10 controls. Levels of IL-8 were significantly increased in vitreous samples from the patients with active PDR (P = .02) when compared with vitreous samples from the controls. The IL-8 levels detected in vitreous samples from patients with inactive PDR were not significantly elevated over those found in the control samples. Interferon-induced protein 10 was detected in the vitreous samples from 23 of 24 patients with active PDR, all patients with inactive PDR, and 9 of 10 controls. Significant elevations of IP-10 were measured in samples from patients with active PDR (P = .004) and in those with inactive PDR (P = .00) over those from controls. In addition, levels of IP-10 were significantly elevated in vitreous samples from patients with inactive PDR compared with vitreous samples from patients with active PDR (P = .02). CONCLUSION: Both IL-8 and IP-10 participate in the pathogenesis of PDR. PMID- 9869786 TI - Effect of brimonidine tartrate on ocular hemodynamic measurements. AB - OBJECTIVE: To study the effect of a selective alpha 2-adrenoreceptor agonist, brimonidine tartrate, on ocular hemodynamics. SUBJECTS AND METHODS: Eighteen patients with ocular hypertension were enrolled in a prospective, randomized, double-masked study in which 0.2% brimonidine tartrate, administered twice daily, was compared with its vehicle in a crossover fashion. The effect on the ocular circulation was assessed by color Doppler ultrasound, which measured blood flow velocities (peak systolic and end diastolic velocities) in the central retinal, ophthalmic, nasal, and temporal ciliary arteries. The following tests were performed at 2 weekly intervals on both treatments, 0.2% brimonidine tartrate and the placebo: intraocular pressure, heart rate, blood pressure, and color Doppler ultrasound. RESULTS: Velocities and resistivity indices measured by color Doppler ultrasound in the ophthalmic artery, central retinal artery, nasal artery, and temporal ciliary arteries showed no statistically significant differences between the placebo and 0.2% brimonidine tartrate when compared with baseline values and between the groups. Intraocular pressure was decreased by 17.7% +/- 9.5% with 0.2% brimonidine tartrate (vs 9% +/- 8% with placebo). CONCLUSION: The hemodynamics of the posterior segment of the eye as measured by color Doppler ultrasound do not appear to be altered by 2% brimonidine tartrate. PMID- 9869788 TI - Does a visible retinal embolus increase the likelihood of hemodynamically significant carotid artery stenosis in patients with acute retinal arterial occlusion? AB - OBJECTIVE: To determine the value of visible retinal emboli as a diagnostic "test" for the detection of hemodynamically significant carotid artery stenosis in the setting of acute retinal artery occlusion. METHODS: A cross-sectional diagnostic accuracy study was performed in a tertiary North American center, with the results of the dichotomous diagnostic test (the presence or absence of visible retinal emboli) being placed against the dichotomous outcome of the presence or absence of hemodynamically significant carotid artery stenosis (defined as > or = 60%, or < 60%, carotid artery stenosis on either side). RESULTS: Forty-eight (18.7%) of our 256 patients had hemodynamically significant carotid artery stenosis. The sensitivity and specificity of retinal emboli for the detection of hemodynamically significant carotid artery stenosis were 39% and 68%, respectively. The presence of a visible retinal embolus generated a likelihood ratio of 1.24 (95% confidence interval, 0.84-1.86). This value corresponds to a patient with a pretest probability of 50% having a posttest probability of 55.3%. The absence of a visible retinal embolus generated a likelihood ratio of 0.88 (95% confidence interval, 0.68-1.15). CONCLUSIONS: The presence of a visible retinal embolus is a poor diagnostic test for the detection of hemodynamically significant carotid artery stenosis in the setting of acute retinal artery occlusion. Accordingly, the presence of an embolus should not influence the decision to perform carotid Doppler ultrasonography in patients with acute retinal arterial occlusion. PMID- 9869789 TI - Availability of color fundus photographs from previous visit affects practice patterns for patients with diabetes mellitus. AB - OBJECTIVE: To determine whether access to color fundus photographs from a patient's previous visit would alter the recommendations rendered to a cohort with diabetic retinopathy. PATIENTS AND METHODS: One hundred sixty patients with diabetic retinopathy returning for a follow-up visit and who had color fundus photographs obtained at a previous visit were evaluated by trained retina specialists. Their clinical impression and recommendations regarding management of diabetic retinopathy were recorded without reference to previous photographs. Color fundus photographs from the patient's most recent visit were then reviewed and new recommendations with regard to appropriate treatment and follow-up were recorded. RESULTS: In 21% of cases, after reviewing the patient's most recent color fundus photographs, the clinical recommendation changed. In 14% of cases, photographs clearly demonstrated that the patient's condition was stable or improved, resulting in a change from recommending treatment to recommending deferral of treatment. In 4% of cases, photographs clearly demonstrated clinical worsening and the recommendation was changed from observation to treatment. In 3% of cases, review of photographs prompted a change in the recommended follow-up interval. CONCLUSION: Access to color fundus photographs from a patient's previous visit frequently changed the clinical recommendations made to patients with diabetic retinopathy regarding appropriate treatment and follow-up. Availability of color fundus photographs therefore has implications about quality of care and may affect the cost of care. PMID- 9869790 TI - The outcome of chemoreduction treatment in patients with Reese-Ellsworth group V retinoblastoma. AB - OBJECTIVE: To determine the outcome of chemoreduction treatment in patients with Reese-Ellsworth group V retinoblastoma. METHODS: Prospective analysis of 27 eyes in 22 patients with group V retinoblastoma treated with either 2- or 6-cycle chemoreduction and focal treatment methods (argon laser photocoagulation, transpupillary thermotherapy, cryotherapy, and plaque radiotherapy). The need for external beam irradiation and the eventual globe salvage rate were assessed. Median follow-up was 28 months. RESULTS: There were 16 eyes in the 2-cycle chemoreduction treatment group and 11 eyes in the 6-cycle chemoreduction treatment group. No significant difference was noted between the 2 groups with respect to baseline patient and eye findings. After chemoreduction treatment, external beam irradiation was necessary in 12 (75%) of 16 eyes in the 2-cycle chemoreduction treatment group and in 4 (36%) of 11 eyes in the 6-cycle chemoreduction treatment group. There was no statistical difference between the 2 and 6-cycle chemoreduction treatment groups with respect to necessity for external beam irradiation (logistic regression analysis). All 4 eyes in the 2 cycle chemoreduction treatment group and 3 of 12 eyes in the 2-cycle chemoreduction treatment and irradiation group were eventually enucleated, the globe salvage rates being 0% and 75%, respectively. Two of 7 eyes in the 6-cycle chemoreduction treatment group and 1 of 4 eyes in the 6-cycle chemoreduction treatment and irradiation group were enucleated, the globe salvage rates being 71% and 75%, respectively. Except for the 2-cycle chemoreduction treatment group, in which the globe salvage rate was significantly lower (P = .03), there was no difference among the other 3 groups (2-cycle chemoreduction treatment and irradiation, 6-cycle chemoreduction treatment, and 6-cycle chemoreduction treatment and irradiation) with respect to globe salvage (logistic regression analysis). CONCLUSIONS: Local tumor control of group V retinoblastoma is possible with 6-cycle chemoreduction and focal therapy when external beam irradiation is not used. A larger sample size is necessary to determine how often external beam irradiation can be avoided. PMID- 9869791 TI - The lacrimal keyhole, orbital door jamb, and basin of the inferior orbital fissure. Three areas of deep bone in the lateral orbit. AB - OBJECTIVES: To calculate the volume of bone in 3 areas of the deep lateral orbit that are available for removal in decompression surgery and to demonstrate these 3 areas within a 3-dimensional computed tomographic reconstruction of the orbit. DESIGN: The 3 areas of bone in the deep lateral orbit were designated the lacrimal keyhole, the sphenoid door jamb, and the basin of the inferior orbital fissure. By means of digitized computed tomographic scans, these 3 areas of bone were analyzed by measuring preoperative and postoperative orbital volumes and predicted bony expansion volumes in 9 patients (17 orbits) who underwent deep lateral orbital decompression surgery. We also calculated the volume of bone that could be removed from 11 normal orbits. A 3-dimensional computer reconstruction of an orbital computed tomographic scan was created, and the 3 areas of potential bone were delineated within it. RESULTS: The average volumes of the basin of the inferior orbital fissure, the sphenoid door jamb, the lacrimal keyhole, and the total of the 3 regions were 1.2, 2.9, 1.5, and 5.6 cm3, respectively. The 3 areas of bone contributed variably to the total, with the door jamb contributing the most volume of the 3, nearly twice the value of the other 2. There was, however, a significant amount of interpatient variability, especially for the door jamb region. CONCLUSION: Orbital decompression surgery of the deep lateral wall can provide adequate volume expansion because of the amount and location of potential space that exists in the 3 areas of deep bone. PMID- 9869792 TI - Ocular levels of azithromycin. AB - OBJECTIVE: To assess azithromycin levels in human serum, aqueous humor, tear fluid, and conjunctival tissue specimens after administration of a single 1-g oral dose of azithromycin. METHODS: Sixty patients undergoing cataract surgery were included in this analysis. Serum, aqueous, and tear specimens were collected 3, 6, and 12 hours and 1, 2, 3, and 4 days after azithromycin administration. Conjunctival tissue biopsy specimens were collected 1, 2, 3, 4, 6, 8, 10, 12, and 14 days after azithromycin administration. All specimens were subjected to analysis by high-performance liquid chromatography-mass spectrometry. RESULTS: Azithromycin concentration ranges during the specified sampling times were as follows: serum, 21 to 974 ng/mL; tear, 82 to 2892 ng/mL; aqueous, 10 to 69 ng/mL; and conjunctival, 0.7 to 32 micrograms/g. Levels above the 90% minimal inhibitory concentration (MIC90) for Chlamydia trachomatis were detected after 4 days in all tear samples and after 14 days in all conjunctival tissue specimens following oral azithromycin administration. CONCLUSION: We demonstrated prolonged high levels of azithromycin in drug-targeted ocular tissue. Prolonged high concentrations of azithromycin in conjunctival tissue make this drug suitable for treatment of conjunctivitis caused by chlamydiae and other susceptible organisms. PMID- 9869793 TI - Advanced glycation end products in age-related macular degeneration. AB - OBJECTIVE: To investigate the localization of N epsilon-(carboxymethyl)lysine (CML), a component and major immunologic epitope of advanced glycation end products, in aged eyes and choroidal neovascular membranes (CNVMs) surgically excised from eyes with age-related macular degeneration. METHODS: Immunohistochemistry for CML was performed using 8 snap-frozen, surgically excised CNVMs. Twelve eyes from patients aged 69 to 82 years and 2 donor eyes, 1 each from a 23-week-old fetus and 21-year-old patient, without age-related macular degeneration or diabetic retinopathy were also examined. To determine if retinal pigment epithelial cells in CNVMs accumulate advanced glycation end products, cytokeratin and CML were stained in paired serial sections. RESULTS: Soft, macular drusen and/or basal laminar and basal linear deposits were observed in 8 of 12 aged eyes. Each case showed CML accumulation, while overlying retinal pigment epithelial cells showed no accumulation in all 12 eyes. In CNVMs, however, retinal pigment epithelial cells showed CML accumulation in their cytoplasm. CONCLUSION: The additional accumulation of advanced glycation end products in soft, macular drusen and/or retinal pigment epithelial cells may play a role in the pathogenesis of CNVM formation in age-related macular degeneration. CLINICAL RELEVANCE: Recently, advanced glycation end products have been found to play a role both in aging changes and neovascularization. Localization of advanced glycation end products in the above-mentioned tissue may lead to a better understanding of the pathogenesis of age-related macular degeneration. PMID- 9869794 TI - Tissue effects of subclinical diode laser treatment of the retina. AB - OBJECTIVE: To determine whether consistent tissue effects are obtained when laser lesions are produced with a commercially available diode laser that are near the limit of clinical detection at the time of treatment. METHODS: Continuous-wave or micropulse diode laser was used to produce clinically undetectable (subthreshold) or barely detectable (threshold) retinal lesions in pigmented rabbits. Tissue effects at intervals after treatment were determined in retinal pigment epithelial (RPE) whole mounts by fluorescence microscopy, and in sections of retina and RPE by light and electron microscopy. RESULTS: Continuous-wave and micropulse laser lesions that were originally clinically undetectable were detectable as zones of pigment mottling after 5 days. By microscopy, compaction and/or swelling was seen in the outer retina, and cells in the RPE monolayer became heterogeneous in size, shape, and pigmentation, but the tissue responses in the outer retina and RPE were variable even within and among lesions in the same eye. CONCLUSIONS: Subthreshold energies used to create both continuous-wave and micropulse laser lesions produced variable effects on the RPE and the overlying neurosensory retina. It appears that, near the minimum effective dose of laser irradiation, individual RPE cell heterogeneity becomes detectable as variability in sensitivity to laser injury. CLINICAL RELEVANCE: As laser energy is reduced to limit collateral tissue damage in clinical applications, it may be difficult to generate reproducible lesions because of heterogeneity among individual cells. PMID- 9869795 TI - Genetic risk of primary open-angle glaucoma. Population-based familial aggregation study. AB - OBJECTIVES: To study familial aggregation of primary open-angle glaucoma in a general population and to determine the absolute and relative risks for first degree relatives. METHODS: First-degree relatives of patients with glaucoma (n = 48) and control subjects (n = 155) from the population-based Rotterdam Study underwent a standardized examination, including perimetry. MAIN OUTCOME MEASURES: Intraocular pressure, vertical cup-disc ratio; and the presence of glaucoma, defined as a visual field defect with a cup-disc ratio of 0.7 or higher or asymmetry of 0.3 or higher between both eyes. RESULTS: The prevalence of glaucoma was 10.4% in siblings of patients, 1.1% in offspring of patients, 0.7% in siblings of controls, and 0% in offspring of controls. Life-time risk of elevated intraocular pressure in relatives of patients vs relatives of controls was 42.5% vs 6.7%, of enlarged cup-disc ratio was 62.2% vs 16.6%, and of glaucoma was 22.0% vs 2.3%, yielding a risk ratio for glaucoma of 9.2 (95% confidence interval = 1.2 73.9). The population-attributable risk of glaucoma was 16.4%. CONCLUSIONS: In a general population, relatives of patients with glaucoma have a strongly increased risk of glaucoma. Enlarged cup-disc ratio, not intraocular pressure, was the earliest and most prominent feature of familial aggregation. Further studies are needed to disentangle the genetic components of the increased familial risk. PMID- 9869796 TI - Genetic risk of age-related maculopathy. Population-based familial aggregation study. AB - OBJECTIVE: To investigate to what extent age-related maculopathy (ARM) is genetically determined. DESIGN AND SETTING: Familial aggregation study based on probands derived from the population-based Rotterdam Study. PARTICIPANTS: First degree relatives of 87 patients with late ARM, i.e., atrophic or neovascular macular degeneration, were compared with first-degree relatives of 135 control subjects without ARM. MAIN OUTCOME MEASURES: Presence and stage of ARM as diagnosed on fundus transparencies, odds ratio, lifetime risk, risk ratio, and population-attributable risk. RESULTS: Independent of other risk factors, the prevalence of early (odds ratio = 4.8, 95% confidence interval [CI] = 1.8-12.2) and late (odds ratio = 19.8, 95% CI = 3.1-126.0) ARM was significantly higher in relatives of patients with late ARM. The lifetime risk estimate of late ARM was 50% (95% CI = 26%-73%) for relatives of patients vs 12% (95% CI = 2%-16%) for relatives of controls (P < .001), yielding a risk ratio of 4.2 (95% CI = 2.6 6.8). Relatives of patients expressed the various features of ARM at a younger age. The population-attributable risk related to genetic factors was 23%. CONCLUSIONS: First-degree relatives of patients with late ARM developed ARM at an increased rate at a relatively young age. Our findings indicate that approximately one fourth of all late ARM is genetically determined and suggest that genetic susceptibility may play an important role in determining the onset of disease. PMID- 9869797 TI - Exposure to allopurinol and the risk of cataract extraction in elderly patients. AB - OBJECTIVE: To determine whether exposure to allopurinol is associated with an increased risk of cataract extraction in elderly patients. METHODS: We conducted a case-control study using data from the Quebec universal health insurance program for all elderly patients. The 3677 cases were patients with a cataract extraction between 1992 and 1994. The 21,868 controls were randomly selected among patients not diagnosed with cataract and matched to cases on the date of the extraction. We determined the odds ratio of cataract extraction according to the cumulative dose and duration of allopurinol use relative to nonusers, using conditional logistic regression analysis. The analysis was adjusted for the effects of age, sex, diabetes mellitus, hypertension, glaucoma, and ophthalmic and oral corticosteroid exposure. RESULTS: A cumulative dose of allopurinol of more than 400 g or a duration of use of longer than 3 years were associated with an increased risk of cataract extraction, with odds ratios of 1.82 (95% confidence interval [CI], 1.18-2.80) and 1.53 (95% CI, 1.12-2.08), respectively. No increase in risk was observed for lower cumulative doses or shorter exposure periods. CONCLUSION: Long-term administration of allopurinol increases the risk of cataract extraction in elderly patients. PMID- 9869798 TI - Laser mania in medicine. PMID- 9869799 TI - Who should perform face-lifts? PMID- 9869800 TI - The quality of medical information on the Internet. A new public health concern. PMID- 9869801 TI - Sunlight exposure and risk of lens opacities in a population-based study. PMID- 9869802 TI - Primary intraocular posttransplantation lymphoproliferative disorder. AB - We report a case of posttransplantation lymphoproliferative disorder manifesting as an isolated, unilateral iris tumor. A 2-year-old boy who had undergone liver transplantation for biliary atresia at age 4 months was seen with a 2-month history of an enlarging iris nodule. Histopathologic examination of the iris lesion demonstrated a mixed population of lymphoid cells. To our knowledge, this is the youngest patient with posttransplantation lymphoproliferative disorder isolated to the eye. PMID- 9869803 TI - Four-eyelid sebaceous cell carcinoma following irradiation. AB - BACKGROUND: Sebaceous cell carcinoma is a distinctive tumor of the eyelid associated with a high rate of metastasis and mortality. Involvement of both upper and lower lids has been described; however, involvement of all 4 eyelids is extremely rare. OBJECTIVE: To describe the evaluation and diagnosis of the clinicopathologic features in a 74-year-old patient with 4-eyelid sebaceous carcinoma and a history of whole face irradiation for eczema. METHODS: Bilateral eyelid, conjunctiva, map, and fine needle biopsy of enlarged, submandibular lymph node specimens were stained with hematoxylin-eosin and oil-red-O for light microscopy and fresh-frozen sections were evaluated by electron microscopy. RESULTS: The biopsy specimens from the eyelids and conjunctiva showed sebaceous carcinoma cells in the meibomian glands and scattered within the conjunctival epithelium. The cells were moderately well differentiated. Similar cells stained with oil-red-O were found in the submandibular lymph nodes. CONCLUSIONS: The appearance of bilateral sebaceous carcinoma supports the concept of simultaneous occurrence of multiple primaries in specific patients, especially after radiation therapy. Patients with a history of facial irradiation and atypical eyelid lesions should be evaluated for malignant tumors such as sebaceous carcinoma. PMID- 9869804 TI - Adenoid cystic carcinoma of the lacrimal gland simulating a dermoid cyst in a 9 year-old boy. AB - Adenoid cystic carcinoma of the lacrimal gland is a malignant neoplasm that is generally found in adults and is usually managed by orbital exenteration and supplemental external beam irradiation or chemotherapy. A recent report has suggested that the tumor may have a less malignant course in children. We describe a case of adenoid cystic carcinoma of the lacrimal gland that simulated a dermoid cyst clinically and radiographically in a 9-year-old boy. The patient was treated with local surgical resection of the mass, followed by orbital plaque brachytherapy. Based on a review of the literature and our recent experience, the advisability of a more conservative approach to this tumor in selected cases is discussed. Although no prognostic conclusions can be drawn on the basis of a single case report with short follow-up, the relatively earlier detection of this tumor made possible by modern orbital imaging studies may allow total removal at an earlier stage and prevent orbital exenteration in a patient with normal vision. Recent developments suggest that there may be a basis for reassessing the advisability of a radical approach to the management of adenoid cystic carcinoma of the lacrimal gland in selected cases. PMID- 9869805 TI - Hemangiopericytoma of the lacrimal sac. AB - Hemangiopericytoma (HPCT) rarely originates in the lacrimal sac; 7 cases have been reported previously and only 1 contained an ultrastructural study. In this article we report an additional case and review the previous reports. While the initial biopsy specimen showed nonspecific cytologic abnormalities, light and electron microscopic studies on the subsequently excised tumor demonstrated that it had a structure characteristic of HPCT. The onset of lacrimal sac HPCT occurs in a younger age group than that of HPCT of other orbital locations. The tumor may recur locally but, to our knowledge, never has been reported to metastasize from a sac location. The treatment goal is complete surgical excision. PMID- 9869806 TI - Lens holder artifact simulating glaucomatous defect in automated perimetry. PMID- 9869807 TI - Increased iris pigment in a child due to latanoprost. PMID- 9869809 TI - Toxocara cati-induced ocular Toxocariasis. PMID- 9869808 TI - New retinoblastoma tumors in children undergoing systemic chemotherapy. PMID- 9869810 TI - A microendoscopic test card for ophthalmic endoscopes. AB - Ophthalmic endoscopes allow examination of intraocular structures when traditional surgical microscopic visualization is limited or impossible. Periodic evaluation of the optical precision of these instruments is essential to ensure optimal intraoperative performance. A test card has been developed to aid in the calibration of ophthalmic endoscopes. Its 5 test targets assess white balance and fiberoptic bundle integrity, focus and resolution, orientation, color fidelity, and angle of view. PMID- 9869811 TI - Ischemic scalp necrosis preceding loss of visual acuity in giant cell arteritis. PMID- 9869812 TI - Iris crystals in Fuch heterochromic iridocyclitis. PMID- 9869813 TI - Neovascularization of the optic disc. What is the origin of the blood flow? PMID- 9869814 TI - Congenital cysts of the iris stroma. PMID- 9869815 TI - Transpupillary thermotherapy in 50 patients with choroidal melanoma. PMID- 9869816 TI - Lesson learned from cataract surgery. PMID- 9869818 TI - 1998 Index to volume 58. PMID- 9869817 TI - Inappropriate use of the term "adenocystic" to refer to "adenoid cystic" carcinoma of the lacrimal gland. PMID- 9869819 TI - The 26th meeting of the International Society for Oncodevelopmental Biology and Medicine, ISOBM 1998. Umea, Sweden, August 30-September 4, 1998. Abstracts. PMID- 9869820 TI - Parachordoma: a clinicopathologic and immunohistochemical study of four cases of an unusual soft tissue neoplasm. AB - Four soft tissue tumors corresponding with the previously reported parachordoma are described. Three of the patients were men, and one was a woman, and their ages ranged from 14 to 53 years (mean age, 29). The tumors were located either superficially or within muscle, and, in one case, involved a tendon. Histologically, the tumors displayed whorls, nests, and pseudoglandular cords of uniform polygonal cells with eosinophilic, vacuolated cytoplasm, in a focally myxoid stroma. Transitions were seen between fascicles of ovoid-spindled cells, with scanty cytoplasm in a fibrous stroma, and, in one case, whorls of bland spindly cells were also present. Electron microscopy of one case showed cells with short interdigitating microvilli and ill-defined junctions. The principal cells in all cases were positive for S100 protein, Leu-7, keratin (CAM5.2), and epithelial membrane antigen (EMA). All tumors were negative with antibody AE1 and with antibodies to cytokeratins CK7 and CK19. No tumor displayed immunoreactivity for carcinoembryonic antigen (CEA), muscle specific actin (MSA), smooth muscle actin (SMA), desmin, glial fibrillary acid protein (GFAP), CD31, or CD34. Parachordoma appears to be an entity with clinical and pathological differences from chordoma, which has a different cytokeratin profile, behaves in a more aggressive fashion, and can dedifferentiate. The differential diagnosis includes myxoid chondrosarcoma, myoepithelial cell tumor, ossifying fibromyxoid tumor, and chondroid lipoma. PMID- 9869821 TI - Nonneoplastic pineal cysts: a clinicopathologic study of twenty-one cases. AB - Twenty-one cases of nonneoplastic pineal cyst are presented. The patients were 13 women and 8 men, with a median age of 33 years. Sixteen patients were symptomatic. Symptomatic cysts had an average size of 16.5 mm. In most cases, symptoms and signs were related to increased intracranial pressure, cerebrospinal fluid obstruction, neuroophthalmologic dysfunction, brainstem and cerebellar compression, and mental status changes. Uncommon clinical presentations in three cases were related to increased cyst size caused by hemorrhage, sudden death, and postural syncope and loss of consciousness. Imaging studies showed a uniform hypodense or hypointense, nonenhancing pineal mass with occasional peripheral calcification and associated with hydrocephalus, aqueductal compression, tectal deformity, and hemorrhage within the cavity, in decreasing order of frequency. Fourteen patients underwent open cyst resection. Histologically, the intact lesions show a unilocular or multilocular cavity, surrounded by a wall comprised of variable amounts of glial tissue, remnants of pineal gland, and an external fibrous capsule. Follow-up information showed 12 patients alive and well without recurrence between 26 and 144 postoperative months. One patient who underwent stereotactic drainage had a recurrence. One symptomatic patient who did not have surgery died suddenly of causes related to the cyst. The present study supports the role of surgical excision for the treatment of symptomatic pineal cysts to obtain adequate tissue for diagnosis and relief of symptoms. The use of histochemical and immunohistochemical studies may prove useful in the distinction of these lesions with astrocytomas and cystic pineal parenchymal tumors. PMID- 9869822 TI - Anaplastic carcinoma of the colon: clinicopathologic study of eight cases of a poorly recognized lesion. AB - Eight cases of a distinctive histological variant of bowel cancer characterized by an anaplastic morphology were identified from 2,650 colonic malignancies (0.3%). The tumors were histologically composed of sheets of anaplastic tumor cells with frequent atypical mitoses, absence of gland formation, and mucicarmine and periodic acid-Schiff (PAS) negativity. Positive immunostaining for cytokeratin and vimentin was observed in eight cases and for epithelial membrane antigen in three; whereas carcinoembryonic antigen, alpha-fetoprotein, S-100 protein, HMB-45 antimelanoma antigen, leukocyte common antigen, and neuroendocrine markers were uniformly negative. Ultrastructural examination demonstrated intercellular tight junctions, focal surface microvilli, and apical terminal webs or long rootlets of microfilaments supporting a colonic derivation. At the time of diagnosis, metastases to regional lymph nodes were found in seven cases and to the liver in six. All patients in this study died of tumor within 9 months. This report emphasizes a poorly recognized variant of colonic carcinoma, characterized by a high degree of anaplasia and malignant behavior. The differential diagnosis for these lesions is discussed. PMID- 9869823 TI - Fibrous cortical defect with bizarre nuclear features. AB - A femoral neck fibrous cortical defect with typical radiographic features had bizarre nuclear features on histological examination. Recognition of the benign radiographic appearance, similar bizarre features of accompanying macrophages, and paucity of mitoses led us to consider this a pseudoanaplastic ischemic or degenerative change in an old fibrous cortical defect. Because the biological behavior was not known with certainty, we closely followed this case. After 25 months, there has been no recurrence or regrowth. The close cooperation between the pathologist and other disciplines facilitates recognition of these pseudoanaplastic lesions and development of treatment strategies for lesions of unknown or uncertain biological behavior. PMID- 9869824 TI - Pathological and radiological correlation of endobronchial neoplasms: Part I, Benign tumors. AB - Benign endobronchial lung neoplasms are extremely rare. Most such neoplasms are of mesenchymal origin; however, neoplasms of submucosal gland origin and surface epithelial origin also occur. The symptomatology and radiographic features of these tumors are often indistinguishable from those of malignant lung tumors. However, recognition and early diagnosis of these lesions may allow for conservative treatment and excellent patient outcome. The clinicopathologic and radiological features of these pulmonary neoplasms are discussed. PMID- 9869825 TI - Amiodarone pulmonary toxicity: cytopathology, ultrastructure, and immunocytochemistry. AB - One hundred ninety cardiac patients were prospectively enrolled in an amiodarone protocol. Over a 10-year period, 16 patients developed new or progressive respiratory symptoms while taking amiodarone. These symptoms included dyspnea associated with abnormal chest radiographs or new or worsening abnormalities on pulmonary function testing. Specimens for microscopic examination were obtained by fiberoptic bronchoscopy with transbronchial lung biopsy (TBB), bronchoalveolar lavage (BAL), open lung biopsy (OLB), or autopsy. Large foamy macrophages with characteristic lamellated cytoplasmic inclusions were noted in all specimens, regardless of other evidence of pulmonary toxicity, suggesting that foamy macrophages represent a routine drug effect. Foamy macrophages were not present in BAL specimens from 53 normal controls and were rarely seen in specimens from 27 patients who had other interstitial lung diseases. When present, the foamy macrophages were less prominent than those seen in specimens from patients receiving amiodarone. Fibrosis was noted in 11 of 16 histological specimens, whereas type II-cell-hyperplasia was observed in 7 of the 16 specimens. Four of the 16 patients with respiratory symptoms died, and their autopsy revealed a combination of foamy macrophages with fibrosis and type II cell hyperplasia reflective of amiodarone pulmonary toxicity. Hyperplastic type II cells were not found in the absence of fibrosis. Immunocytochemistry allowed differentiation between foamy macrophages and type II cells and represents a useful tool for future investigations of the pathogenesis of amiodarone-induced pulmonary disease. PMID- 9869826 TI - Pathology of human immunodeficiency virus infection: noninfectious conditions. AB - Diagnostic anatomic pathologists play a crucial role in the battle against acquired immunodeficiency syndrome (AIDS). Not only are they intimately involved in the treatment of individual patients with human immunodeficiency virus (HIV) infection, but also they make important observations that result in the expansion of the scientific understanding of its pathogenesis. Pathologists studying tissue from patients with HIV infection should be familiar with the conditions to which these patients are susceptible. Although opportunistic infections are important causes of morbidity and mortality, noninfectious conditions frequently make substantial contributions to the disease course. Patients with HIV infection may be at increased risk for neoplastic disease. They do not, however, have an increased incidence of the most common tumors affecting the general population, such as breast, colon, and prostate carcinoma. Immunodeficiency results in increased susceptibility to malignant neoplasms, both by decreased immunologic response to abnormal cells and increased susceptibility to infection by viruses. All of the malignant neoplastic diseases that are Centers for Disease Control and Prevention (CDC) AIDS indicator conditions have been shown to have an association with a virus: Kaposi sarcoma (KS) with herpes hominis virus 8 (HHV-8), malignant lymphoma with Epstein-Barr virus (EBV), and cervical carcinoma with human papilloma virus (HPV). Patients with HIV infection also can develop reactive processes that are attributable to direct effects of HIV or immune system alterations. Such conditions include salivary gland cystic lymphoepithelial lesion, lymphadenopathy, lymphocytic interstitial pneumonitis, encephalopathy, enteropathy, nephropathy, hepatic conditions, dermatologic conditions and anemia. PMID- 9869827 TI - Rudolf Virchow, not just a pathologist: a re-examination of the report on the typhus epidemic in Upper Silesia. AB - Rudolf Virchow (1821-1902) is mainly remembered as the "father" of cellular pathology; however, he was not just a pathologist. His contributions to anthropology, archeology, ethnography, and history, as well as his involvement in epidemiology, public health, and politics, portray a man with multiple interests, deeply engaged in the controversies of his time. In his Report on the Typhus Epidemics of Upper Silesia of 1848, the young Virchow reveals himself to be a self-assured pathologist, although his postmortem examinations failed to shed much light on typhus. Despite of his shortcomings and biases, Virchow's genius is revealed in his deep appreciation of the importance of the total physical, socio cultural, economic, and political background of epidemic diseases. One can discern in the Report the making of Virchow, the politician and statesman who will contribute to the modernization of Germany's public health, and of the physician-scholar and physician-citizen who, despite of his shortcomings and militancy, continues to inspire and challenge us today. PMID- 9869828 TI - Ossifying fibromyxoid tumor of soft parts: a report of 17 cases with emphasis on unusual histological features. AB - Ossifying fibromyxoid tumor of soft parts is an unusual benign neoplasm, with a tendency for local recurrence. Its typical microscopic appearance is that of a multinodular proliferation of round to spindle shaped cells separated by fibrous bands in which bone formation is often seen. Herein, we present the clinicopathologic features of 17 examples of this tumor with particular emphasis on some unusual histopathologic features that may place pitfalls in the diagnosis of this tumor, including satellite micronodules, mucinous microcysts, absence of myxoid areas, crush artifact, multiple microcalcifications, epidermoid cysts, atypical chondroid differentiation with binucleate lacunar cells, pericytic growth pattern, and malignant change. Awareness of these unusual morphologic features should lead to a search for areas displaying the more typical features of ossifying fibromyxoid tumor to arrive at a correct diagnosis. PMID- 9869829 TI - Postoperative granulomas of the endometrium: histological features after endometrial ablation. AB - Postoperative granulomas of the male urogenital tract are a well-recognized phenomenon. Similar granulomas have also been described in the uterine cervix, fallopian tube, and other sites after various procedures, as well as in the endometrium after endometrial ablation procedures. Endometrial ablation is a procedure increasingly used by gynecologists to relieve symptoms associated with dysfunctional uterine bleeding. Occasionally, patients will not have a satisfactory result, and some will require subsequent hysterectomy. We describe the pathological findings in the hysterectomy specimens from 15 patients who had previously undergone endometrial ablation. Indications for subsequent hysterectomy included dysmenorrhea (7 patients), menorrhagia (7 patients), dysfunctional uterine bleeding (5 patients), and pelvic pain (4 patients). All patients had varying degrees of fibrosis of the endometrial cavity, with some endometrial cavities completely obliterated by fibrous tissue. Histological examination revealed fibrosis with varying degrees of granulomatous inflammation. The majority of the granulomas were associated with refractile brown hematoidin like pigment, and most were also associated with uniform black pigment. In 8 cases, areas of faintly eosinophilic, homogenous, hyalinized material were present within the endometrium. Comparison is made to granulomas due to other causes, because the postoperative granulomas of the endometrium differ morphologically from granulomatous inflammation caused by other etiologies. As endometrial ablation gains popularity among gynecologists and their patients, it is likely that the practicing pathologist may encounter these sequelae with increasing frequency. PMID- 9869830 TI - Collision tumor: a peripheral neuroepithelioma and a transitional-cell carcinoma occurring simultaneously in the renal pelvis. AB - We report a unique case of a collision tumor of the kidney composed of a peripheral neuroepithelioma (PNE) and a transitional cell carcinoma (TCC) that occurred in a 67-year-old man with a long history of superficial TCC of both the bladder and bilateral upper urinary tracts. The PNE was composed of hyperchromatic, small, round cells, arranged in clusters and poorly cohesive sheets. Immunohistochemical studies revealed the small cells to be strongly positive for O13 (ie, a MIC2 gene product stain [CD99]) and vimentin. Electron microscopic examination found numerous interdigitating cell processes, multiple poorly formed cell-cell junctions, and dense-core granules. The latter were most conspicuous at the tips of the cell precesses. The PNE cells were clearly negative for cytokeratin, and they showed no transitional or intermediate features that would suggest evolution of the PNE from the adjacent TCC. The proximity of these two tumors appears to be serendipitous; thus, it can be considered a true collision tumor of the kidney. PMID- 9869831 TI - The Hermansky-Pudlak syndrome. AB - Hermansky-Pudlak Syndrome (HPS) is a rare, inheritable disorder characterized by the classic triad of oculo-cutaneous albinism, platelet dysfunction, and ceroid deposition. An associated complication is pulmonary fibrosis with progressive restrictive lung disease. This report discusses the lung involvement often seen in this condition correlated with radiography, computed tomography, high resolution computed tomography, and the underlying pathology, by means of two such afflicted siblings. The elder died of respiratory failure while awaiting lung transplantation. The younger sibling is currently undergoing evaluation for transplantation. PMID- 9869832 TI - Minimal criteria for the diagnosis of prostate cancer on needle biopsy. AB - Increased clinical screening of men at risk for prostate cancer, and the realization of the benefits of performing multiple biopsies per prostate, have facilitated early detection of malignancy, while presenting the pathologist with a growing array of diagnostic findings. Interpretation of these findings requires discussion of the minimal criteria required for the diagnosis of cancer on needle biopsy within a wide spectrum of related histologic findings. This spectrum includes small acinar proliferations suspicious for but not diagnostic of cancer, benign mimics of cancer, the preinvasive entity of high-grade prostatic intraepithelial neoplasia, and various treatment effects. Clinical implications of these findings and other prognostic factors are detailed. PMID- 9869833 TI - On writing medical history. AB - Writing medical history papers can be professionally and personally rewarding, but a little overwhelming for novices. This paper offers step-by-step guidelines for the entire process, including developing appropriate topics, finding and using appropriate sources, and writing the paper. Specific reference sources are recommended. PMID- 9869834 TI - Immunologic barriers in pig-to-primate cardiac xenotransplantation. PMID- 9869835 TI - Musings on intrathoracic transplantation past and future. PMID- 9869836 TI - Lysis of pig endothelium by IL-2 activated human natural killer cells is inhibited by swine and human major histocompatibility complex (MHC) class I gene products. AB - We have previously described a form of xenograft rejection, mediated by natural killer (NK) cells, occurring in pig-to-primate organ transplants beyond the period of antibody-mediated hyperacute rejection. In this study, two distinct NK activation pathways were identified as mechanisms of pig aortic endotheliual cell (PAEC) lysis by human NK cells. Using an antibody-dependent cellular cytotoxicity (ADCC) assay, a progressive increase in human NK lysis of PAEC was observed following incubation with human IgG at increasing serum titer. In the absence of IgG, a second mechanism of PAEC lysis by human NK cells was observed following activation with IL-2. IL-2 activation of human NK cells increased lysis of PAEC by over 3-fold compared with ADCC. These results indicate that IL-2 activation of human NK cells induces significantly higher levels of lytic activity than does conventional ADCC involving IgG and FcRIII. We next investigated the role of MHC class I molecules in the regulation of NK lysis following IL-2 activation. PAEC expression of SLA class I molecules was increased by up to 75% by treatment with human TNFa. Following treatment with TNFa at 1 u/ml, IL-2 activated human NK lysis of PAEC was inhibited at every effector:target (E:T) ratio tested. Maximal effect occurred at an E:T ratio of 10:1, with TNFa inhibiting specific lysis by 59% (p < 0.01). Incubation with an anti-SLA class I Mab, but not IgG isotype control, abrogated the protective effects of TNFa on NK lysis of PAEC, suggesting direct inhibitory effects of SLA class I molecules on human NK function. To investigate whether human MHC class I molecules might have similar effects on human NK lysis of PAEC, further experiments were performed using a soluble peptide derived from the alpha-helical region of HLA-B7. Incubation with the HLA B7 derived peptide significantly reduced the IL-2 activated NK lytic activity against PAEC in a dose-dependent fashion. Maximal effect occurred at a concentration of 10 mg/ml, where an 8-fold reduction in IL-2 augmented NK lysis was observed (p < 0.01). These results suggest that IL-2 activated human NK lysis of porcine xenografts may be inhibited by strategies which increase PAEC expression of SLA class I molecules, introduce HLA class I genes into PAEC, or use soluble HLA class I peptides. PMID- 9869837 TI - Human NK cells demonstrate increased adherence and lysis of porcine endothelium following activation with IL-2. AB - BACKGROUND: For clinical xenotransplantation of pig organs to be successful, the significant barriers of host cellular immunity against the pig will need to be overcome. Natural killer (NK) cells appear to play an important role in rejection of discordant xenografts, and IL-2 activated human NK cells are cytotoxic to human endothelium. We addressed the role of IL-2 activated human and baboon NK cells in the primate immune response to pig xenoantigens in vivo and in vitro. METHODS: Human natural killer (NK) cells were isolated from pooled human peripheral mononuclear cells (PBMC) using density gradient centrifugation method. Phenotype of isolated cells was confirmed by flow cytometry. Purified NK cells were than tasted for spontaneous and IL-2 augmented adhesion and lysis of cultured pig aortic endothelial cells (PAEC) monolayers in a short-term 51 Cr release assay. The effects of human macrophage derived cytokines TNF alpha and IL 1 on adhesion of IL-2 activated NK cells were studied. RESULTS: Human NK cells demonstrate prominent adherent and lytic activity against pig endothelium, which can be significantly augmented by IL-2. Macrophage derived cytokines can further augment the adherent properties of these NK cells. CONCLUSIONS: These result suggest that human NK cells are likely to play a significant role in rejection of pig-to-human xenografts. PMID- 9869838 TI - Demand, supply and allocation in Eurotransplant. AB - The main problem in organ transplantation is the continuing shortage of organ donors. Despite all efforts no major significant increases in organ availability are observed during the year 1996 in the participating Eurotransplant counties, while the demand i.e. the waiting lists are still increasing. Shortage of organs will also have its effects and constraints on the distribution i.e. the allocation of scarce organs. To meet the demand of the renal transplantation programs a special kidney allocation system was designed based upon many simulation studies. Already a few months after implementation of the new system very promising results were observed i.e. the discrepancies between the different countries in terms of kidney procurement and transplantation frequencies disappeared. Furthermore, twice as much long waiting kidney patients have been transplanted as previously and the percentage of well matched HLA donor-recipient combinations remained surprisingly high, nearly 24%. PMID- 9869840 TI - Comparison of serological and molecular (PCR-SSP) techniques of HLA-DR typing in clinical laboratory routine. AB - Advances in molecular biology techniques allowed for introduction of PCR-based methods for HLA typing. In routine HLA typing for organ transplantation serological method is still being used as a standard, although molecular techniques are applied more and more often. The aim of our study was to compare HLA-DR typing using traditional serological method and PCR-SSP methodology in routine clinical laboratory. HLA-DR typing was performed using standard microcytotoxicity assay and PCR-SSP method in 28 patients referred to our Transplantation Immunology Unit for HLA typing. Comparison of results obtained by both methods revealed no discrepancies in 5 patients, in 12 patients the PCR-SSP typing showed additional DR antigens or splits of antigens. In 11 patients serological typing turned out to be impossible because of technical problems. Molecular PCR typing allowed for precise antigen determination in all the patients. Comparing both methods we found PCR-SSP HLA typing method very useful in routine HLA-DR determination, especially valuable in patients, in whom some problems in serological testing are expected. PMID- 9869839 TI - The effect of calcium in the UW solution on preservation of the rat liver. AB - In this study we investigated the effect of calcium addition to the UW solution on the quality of the preserved rat liver as judged by normothermic isolated perfusion. Rat livers were cold stored in UW solution containing varying concentrations of calcium chloride (0, 0.5, 1.5, 5.0 mM) for periods of 0, 24 and 48 hours. At the end of the preservation period the livers were reperfused for 90 minutes at 37 degrees C with Krebs Henseleit Buffer. The quality of preservation was assessed by quantification of enzyme release, bile production and protein synthesis. The addition of 1.5 mM calcium to the UW solution suppressed the incidence of damage in the 24 hour cold stored liver similar to control livers (0 hours preserved). LDH release were significantly reduced from 22.1 +/- 7.3 (units/hr/g) in regular UW to 9.4 +/- 0.8 (units/hr/g) in UW plus 1.5 mM calcium. AST release also was suppressed by the addition of calcium to the UW. Bile production was enhanced by the addition of calcium; from 21.3 +/- 0.6 (mg/hr/g) in regular UW to 46.3 +/- 5.9 (mg/hr/g) in UW plus 1.5 mM calcium. Protein synthesis was reduced to 38% of control after 24 hr cold storage and was unchanged by the addition of calcium to the preservation solution. Although the addition of calcium to the UW solution improved the preservation of the 24 hour cold stored liver it did not offer the same degree of protection to the 48 hour preserved liver. Therefore, calcium addition may be one agent for improving preservation for short term cold storage of the liver but longer term storage will require other modifications as well. PMID- 9869841 TI - Biological skin covers in treatment of two cases of the Lyell's syndrome. AB - The treatment of two cases of toxic epidermal necrolysis (Lyell's syndrome) is described. Although some features were common for both ones (young men practically of the same age, reaction after using the same drug) the clinical course of illness was very different. Spontaneous epithelisation of partial thickness lesions and definitive healing under the xenografts in one patient and full-thickness skin-loss on 12% of body surface with severe septic complications requiring application of cultured keratinocytes and/or skin autografting in the other patient were the main differences. The interdisciplinary approach using a burns treatment protocol in non-burned patient including the close co-operation with the tissue bank in preparing different types of biological covers has been applied. PMID- 9869842 TI - Allogenic preserved costal cartilage in reconstructive surgery. AB - Allogenic, preserved cartilage is often used for reconstruction of face. This study was undertaken to analyze the effects of cartilage transplantation. In analyzed group of 437 patients after cartilage transplantation, 42.2% were operated because of posttraumatical changes, 29.0% because of congenital malformations. In 16.7% nonspecific inflammations were the cause of reconstructive operations. Malformations were mainly localised in nose 59%, ear concha 16.5% and mandible 10.9%. Human costal cartilage, preserved in 0.9% NaCl and radiation-sterilized was used for reconstruction. 24-190 months after surgery (in several clinical units) patients were examined and results were collected in special questionnaire by the team who performed surgery. The results of treatment were compared with age, diagnosis and localisation of changes. It was found that very good result of treatment was achieved in 33.5% of patients, in 41.8% result was satisfactory. In 19.9% of operated patients result of treatment was unsatisfactory. Correlation of some clinical and biological characteristics with the result of treatment is discussed. PMID- 9869843 TI - Electron microscopic study on the effects of different preservation methods for meniscal cartilage. AB - OBJECTIVES: The study was conducted to evaluate different methods for preservation of meniscal cartilage. During the last decade, the use of allografts containing cartilage in various forms, such as: small osteochondral grafts, osteoarticular grafts, or whole joints, has gained popularity. Meniscal allografts in particular, can be used in cases of post-menisectomy syndrome, or in uni-compartmental osteoarthritis of the knee. The preservation methods used for the cartilaginous components of allografts would appear to influence the success rates, which vary between 30% and 85% according to different reports. Some of the methods of preservation currently in use include: deep-freezing in liquid nitrogen, freezing to -80 degrees C, gamma irradiation, and immersion in ethylene oxide. METHODS: In an attempt to find the best technique for cartilage preservation (in terms of preserving the ultra-structural properties prior to implantation), bovine meniscal cartilage was examined after preservation by: a) freezing to-80 degrees C; b) exposure to 2.5 Mrad gamma irradiation; or c) a combination of both. RESULTS: Electron microscopic studies were performed, with fresh menisci serving as controls. Our tests demonstrated that in all techniques, severe cellular damage was caused, whereas the collagen network retained its properties. CONCLUSION: The optimal method for the preservation of cartilage remains to be established. PMID- 9869844 TI - Human immunodeficiency virus (HIV) inactivation of banked bone by gamma irradiation. AB - OBJECTIVE: The increasing number of patients with acquired immunodeficiency syndrome (AIDS) and human immunodeficiency virus(HIV)-positive carriers, poses difficulties when musculo-skeletal tissues are considered for banking in readiness for future clinical application. This study was conducted to test the actual yield of gamma irradiation on HIV infectivity, within HIV-infected bones. METHODS: The effect of gamma irradiation on bones containing T-cells chronically infected with HIV type I (HIV-I) was studied, in respect to inactivation of the virus. RESULTS: After exposure of the cell-free virus or infected T-cells to 2.5 megarads of gamma irradiation, the authors were able to demonstrate complete inactivation of the virus. CONCLUSIONS: It would appear from this study that gamma irradiation at this dose is sufficient to achieve clinical sterilisation of bones and facilitate their use for reconstructive procedures by eliminating the risk of HIV transmission to the recipient. Furthermore, when preparing bones for banking, this would also seem to be the method of choice in preventing the transmission of various strains of bacteria, fungi and other viruses. PMID- 9869845 TI - Bone-implant interface, objectivization of X-ray microanalysis. AB - Several techniques of the bone-implant interface uncovering in the proper way for X-ray microanalysis, were tested. The obtained results confirmed that the method of sample preparation has significantly influenced the results of investigation. The mechanical polishing, which is routinely used for preparation of surfaces to the X-ray microanalysis, introduces artifacts. Cutting by means of microtome, in the case of brittle implants, and sawing by utilizing diamond saw, in the case of metallic ones, were indicated as the optimal methods. Cleaning of the samples in the ultrasonic cleaner before covering them with conductive layer is required. PMID- 9869846 TI - Long-term follow up after transplantation of frozen and radiation sterilized bone grafts. AB - The application of frozen and radiation sterilized allogenic bone grafts for reconstructions in orthopaedic operations is described. Analysis of results of treatment of 1125 patients was performed. It was found that use of preserved bone allows to reduce the extend and duration of surgery. Nearly total substitution of grafts may be seen in 3 to 8 months after surgery. PMID- 9869847 TI - An optimization study on unprogrammed cryopreservation of human CD34+ bone marrow cells and their subsequent storage in an -80 degrees C mechanical freezer. AB - Among the factors which enable successful bone marrow transplantation, the ability to store and subsequently recover sufficient viable marrow cells to reestablish hematopoiesis is crucial. In most centers, marrow is typically prepared for long term storage by programmed, controlled rate freezing with subsequent placement into the vapor phase of liquid nitrogen. Though clearly effective, this procedure is time consuming and equipment intensive. Since cost containment is an important issue for all medical procedures, we evaluated the utility of a faster and less costly method to accomplish the same purpose. We found that when CD34+ bone marrow cells are immediately frozen and stored at -80 degrees C, the number and quality of stem/progenitor cells recovered after thawing was comparable to those reported recovered after conventional processing. Herein we report optimized conditions for cryopreserving and storing CD34+ bone marrow cells at -80 degrees C and for subsequent thawing of the stored product. With these methods post-thaw recovery of CFU-Mix, for example, equaled or exceeded 80% of predicted numbers, even after six months of storage. Further, though progenitor cell recovery does not necessarily correlate with speed of engraftment, these results nonetheless suggest that storage of human CD34+ cells at -80 degrees C in a mechanical freezer is a convenient, inexpensive, and reliable method for storing marrow for transplantation. PMID- 9869848 TI - Higher exposure to cyclosporine A with unchanged tolerability in patients converted from Sandimmun to Sandimmun Neoral. AB - Neoral (NEO) is claimed to have better pharmacokinetics than standard preparation of cyclosporine (SIM) thus providing more reliable immunosuppression. We estimated safety and tolerability of NEO and compared pharmacokinetic parameters in 20 stable renal allograft recipients (RARs) converted from SIM to NEO treatment. Another 20 stable RARs continuously treated with SIM created a control group. Whole blood through CsA level (C0) did not differ after conversion (SIM: 136.2 +/- 33 ng/ml and NEO: 142.6 +/- 34 ng/ml). During therapy with NEO peak blood concentration (Cmax) was significantly higher (935.6 +/- 368 ng/ml) and occurred earlier (Tmax 1 hr. 36 min. +/- 30 min) as compared to the period on SIM (Cmax 598 +/- 309 ng/ml, p = 0.01), Tmax = 3 hr. +/- 1 h 36 min., (p = 0.01) respectively. AUC increased from 2975.4 +/- 1020 ngxhr/ml to 4236.1 +/- 1188 ngxhr/ml (p < 0.0001). Correlation coefficient between AUC and C0 was higher during NEO (r = 0.52) than SIM therapy (r = 0.32). The only noticeable change in laboratory tests after switch to NEO was slight increase of serum triglyceride concentration (119.5 +/- 44.7 mg/dL vs. 148.4 +/- 67.0 mg/dl). The mean serum creatinine concentration did not change significantly (1.42 +/- 0.32 mg/dL and 1.46 +/- 0.31 mg/dL). Tolerance of NEO was good and 1:1 switch from SIM to NEO is clinically safe. Higher bioavailability of NEO was not associated with decreased tolerability or increased nephrotoxicity. Better correlation between C0 and AUC during NEO administration makes CsA treatment monitoring more reliable. PMID- 9869849 TI - Cellular adhesion molecules changes in myocardium during first year post heart transplant. AB - The increased presence of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1) in the myocardium after orthotropic heart transplantation (OHT) has been implicated in early and late organ rejection. The aim of this study was to investigate the changes in ICAM-1 and VCAM-1 during the first years post-OHT. Accordingly, we studied multiple endomyocardial biopsy specimens collected from 11 randomly selected patients (all males, mean age 46 +/ 11 years). Qualification criteria for OHT included: ischemic cardiomyopathy in 6 pts (55%) and idiopathic dilated cardiomyopathy in 5 pts (45%). All patients were receiving triple immunosuppressive regimen. The therapy consisted of cyclosporine, azathioprine, and prednisolone. Multiple endomyocardial biopsy specimens were collected at 7, 30, 90 and 360 days post OHT from all patients (pts), and examined routinely for histologic signs of organ rejection. To assess levels of adhesion molecules we used monoclonal antibodies (murine anti-human ICAM-1 and VCAM-1) on frozen sections. Immunoreactivity (IR) was detected using a commercially available kit. Intensity of IR was assessed based on a semiquantitative scoring system. In this study, IR scores > or = 2 + were considered positive for ICAM-1, and scores > or = 1 + were considered positive for VCAM-1. IR scores in specimens obtained from consecutive biopsies were compared with the initial biopsy collected at the day 7. The results were analyzed using nonparametric statistics. The routine evaluation revealed histological signs of organ rejection (> or = 2) in 2 pts at 7 days, in 5 pts at 30 days, in 3 pts at 90 days, and in 1 patient at 360 days. On the other hand, ICAM-1 and VCAM-1 expression were absent in the majority of patients at 7, 30 and 90 days, but their presence was significantly increased at 360 days (p < 0.05). Absence of the early expression of ICAM-1 and VCAM-1 may be related to the protective effect of triple immunosuppressive therapy in these patients. The expression of ICAM-1 and VCAM-1 strongly emerging at 1 year post OHT may reflect chronic rejection in myocardium. In conclusion, the immunohistological monitoring of the adhesion molecules in biopsy specimens during routine biopsy schedule may be helpful for the discovery of chronic rejection. PMID- 9869850 TI - Reversal of hyperglycemia in streptozotocin diabetic mice by xenotransplantation of microencapsulated rat islets. AB - Rat pancreatic islets were immunoisolated within alginate capsules with additional polyethyleneimine-protamine-heparin highly biocompatible membrane. Perifusion study in vitro demonstrated satisfactory similarities between the insulin release profiles of encapsulated and free islets. Concordant xenotransplantation of microencapsulated rat islets significantly prolonged mean time of restored normoglycemia (46 +/- 15 days) in streptozotocin-diabetic BALB/c mice recipients comparing to uncoated grafts (7 +/- 2 days). PMID- 9869852 TI - Antibodies and chronic organ graft rejection. PMID- 9869853 TI - Chronic rejection from bedside to bench: role of T cells. PMID- 9869854 TI - Allograft coronary disease: the role of antigen independent mechanisms in pathogenesis. PMID- 9869855 TI - Non-immune factors in chronic rejection. Is there a role for hypolipemic drugs? PMID- 9869856 TI - Liver graft bile duct necrosis--indication for retransplantation. AB - The authors describe the first in Poland case of liver retransplantation in an adult recipient, in whom the indication for the procedure was bile duct necrosis following liver transplantation due to liver cirrhosis of an uncertain origin. PMID- 9869857 TI - Beginning of pancreatic islet isolation by collagenase digestion (personal reminiscences). AB - The beginning of pancreatic islet isolation by collagenase digestion is described in the form of personal account. PMID- 9869858 TI - Monitoring human islet allografts using a forearm biopsy site. AB - OBJECTIVES: Despite the increasing success of whole-organ pancreas transplantation, the success of clinical islet allografts has remained limited. One of the factors limiting the success is the difficulty in monitoring islet allografts after transplantation. The aim of these studies is to develop a method of "biopsying" human islet allografts using a forearm islet implantation site. METHODS: A subtherapeutic number of isolated human islets were placed in the forearm under the muscle fascia in three human recipients with Type I insulin dependent diabetes. All of the recipients had undergone successful cadaveric renal transplantation at least one year prior and were maintained on their baseline immunosuppression. Aliquots of the islet grafts were removed 7 and 14 days to assess engraftment and graft infiltrate. To verify that the islets were viable, 400 were handpicked and transplanted into B6-scid mice made diabetic with streptozotocin. RESULTS: The biopsy site was found in all three cases. In one patient, no islets were recovered. In two other patients, viable islet tissue was recovered 7 days after transplantation. Immunohistology at 7 days showed the presence of both insulin and glucagon-staining cells in the islets. At 14 days in these two patients, a mononuclear cell infiltrate was observed in the explanted islet biopsies. Immunohistology showed the relative absence of insulin-staining cells with intact glucagon-staining cells. This finding is consistent with recurrent autoimmunity in the islet grafts. DISCUSSION: This preliminary study shows that the forearm biopsy site is a useful method to retrieve human islet grafts after transplantation. The islets engraft and are easily found in the first weeks after transplantation. These data suggest that recurrent autoimmunity may affect islet allografts. Further studies will be needed to determine if the histology in the forearm will correlate with the fate of intraportal or intraperitoneal islet allografts. Although they were shown to reduce the incidence of early allograft failure, their influence on the long-term graft survival remains to be proven. PMID- 9869859 TI - Immunophenotyping fish-to-mouse islet xenograft rejection: a time course study. AB - Tilapia islets, Brockmann bodies (BBs), transplanted under the kidney capsule (KC) of diabetic nude mice provide long-term normoglycemia, but, when transplanted into euthymic mice, reject in about one week. OBJECTIVES: The present study characterizes the cellular infiltrates at several time points during the xenograft rejection process. METHODS: Tilapia BBs were harvested, fragmented, cultured overnight, and then transplanted under the KC of streptozotocin-diabetic Balb/c mice. Glucose levels were measured daily until the mice were killed at 1 (n = 2), 2 (n = 2), 3 (n = 3), and 5 days (n = 3) post transplantation and at the time of BB graft rejection (n = 6). Serial frozen sections of graft-bearing kidneys were stained for murine macrophages (MOMA-2, F4/80, M170), CD4+ (L3T4) T-cells (YTS 191.1), and CD8+ (Ly-2) T-cells (YTS 169.4) by indirect immunoperoxidase; the presence of granulocytes and plasma cells was assessed with H&E stained sections. RESULTS: At 1 day, the grafts have undergone some central necrosis with macrophage infiltration. By 2 days, these changes are very well-developed and granulocytes, almost exclusively eosinophils, begin to surround the graft. At 3 days, rare CD4+ and CD8+ T-cells are seen at the graft kidney interface. Macrophages massively infiltrate the necrotic foci and pepper the graft. At 5 days and at rejection, macrophages and eosinophils predominated in the center of rejecting grafts while CD8+ T-cells and CD4+ T cells were present at the periphery. Plasma cells were rare. CONCLUSION: We conclude that cell-mediated processes and eosinophils play roles in the rejection of cellular xenografts across this very wide phylogenetic barrier. PMID- 9869860 TI - Expression pattern of extracellular matrix proteins in the pancreas of various domestic pig breeds, the Goettingen Minipig and the Wild Boar. AB - In spite of progress in biotechnology, isolation of porcine pancreatic islets remains a difficult task with unpredictable results. One reason could be the lack of knowledge as to the expression of extracellular matrix proteins in porcine exocrine and endocrine tissues, particularly in "islet capsules". Such proteins are subject to digestion by proteases, yet they might have a protective function for the fragile islets. OBJECTIVE: Of our study was a detailed histological analysis of the extracellular matrix proteins in various pig breeds. A broad panel of commercial, human-specific antibodies were used, since antibodies against porcine tissue were not available. METHODS: Frozen pancreatic tissue section of 7 domestic pig breeds, the Goettingen Minipig and the Wild Boar were stained with antibodies against collagen types I, II, III, IV, V, VI, VII, IX, laminin, fibronectin, vitronectin and elastin. Binding of antibodies was detected by immunoperoxidase and evaluated microscopically. Human and rat tissue was treated in the same way. RESULTS: (1) With the exception of anti-collagen type II, type VII and vitronectin, all antibodies revealed distinct binding patterns in the pancreas of the different pig breeds. However these antibodies bound on human cartilage and skin. (2) Collagen types I, III, IV, laminin and fibronectin are expressed on porcine pancreatic "islet capsules". (3) Expression levels of these proteins on "islet capsules" vary in the different pig breeds. However, no significant differences could be found in the expression pattern of collagen types I, III, IV, laminin and fibronectin, comparing domestic, experimental and wild type pigs. (4) Older individuals (Goettingen Minipig) appear to express higher levels of proteins on "islet capsules" than younger ones. CONCLUSIONS: Antibodies with specificity for human extracellular matrix proteins can be used successfully in the porcine pancreas. Thus, analysis of the structure and composition of porcine pancreatic tissue can be performed even without pig specific antibodies. Particularly, the effects of various proteases and collagenases on the pancreatic tissue can now be monitored by immunohistochemical analysis allowing a rational design of protease mixtures for the isolation of pancreatic islets. PMID- 9869861 TI - Evaluation of immunoisolated insulin-secreting beta TC6-F7 cells as a bioartificial pancreas. AB - OBJECTIVE: To evaluate the growth and insulin secretion from microencapsulated beta TC6-F7 cells in vitro and to assess the in vivo function of microencapsulated cells transplanted in rats with steptozotocin (STZ)-induced diabetes. METHOD: Alginate-poly-L-lysine encapsulated beta TC6-F7 cells were exposed to glucose, isobutylmethylxanthine (IBMX) and glucagon-like peptide I (7 36 amide) in a static in vitro challenge. In vivo, 2.5-3.5 x 10(7) encapsulated cells were implanted into diabetic rats. Graft function was evaluated by monitoring blood glucose concentrations and by an intraperitoneal glucose tolerance test. RESULTS: The cell density (number of cells per capsule) of cultured microencapsulated beta TC6-F7 cells increased almost 35-fold over a 55 day observation period to reach a plateau of approximately 3500 cells/capsule. While insulin secretion per capsule remained unchanged over the first 21 days of culture, a 7-fold increase was observed during the last 14 days of the 55 day observation period. Intraperitoneal transplantation of 3.5 x 10(7) encapsulated cells into diabetic rats resulted, within 24 hours, in reversal of hyperglycemia for up to 60 days. Post-transplantation blood glucose concentrations varied between 2 and 4 mM. Glucose clearance rates evaluated by an intraperitoneal glucose tolerance test at 30 days post-transplantation resulted in a markedly flat glucose clearance curve with blood glucose never rising above 4 mM. The glucose challenge of microencapsulated cells recovered 30 days post transplantation resulted in a 2-fold increase in insulin response at glucose concentrations greater than 5.5 mM as compared to glucose-free media. In addition, immunostaining of recovered grafted tissue for insulin, reveals a strong presence of the peptide within the cell population. CONCLUSIONS: These data demonstrate the potential use of an immunoisolated beta-cell line for the treatment of diabetes. PMID- 9869862 TI - Characterization and transplantation of agarose microencapsulated canine islets of Langerhans. AB - The bioartificial pancreas was designed to incorporate islet tissues and a selectively permiable membrane that isolates islets from the immune system of the recipient. The efficacy of agarose, a nontoxic polysaccharide, has been evaluated as a material of microcapsules to prevent allo- and xenograft rejection in rodents. The aim of this study is to demonstrate the possibility of the agarose microcapsule containing allo-islets as a bioartificial pancreas in canine model. In vitro viability of islets was determined by glucose challenge during perifusion experiments (n = 4). Insulin secretion from both encapsulated (enc.) and non-encapsulated (non-enc.) canine islets rose from initial basal levels of 0.09 (encap.), 0.07 (non-encap.) to the peak of 0.2 (encap.), 0.1 (non-encap.) in microU/islet/min after 5 minutes, then decreased to the basal level when the glucose challenge was discontinued. Auto-transplantation was performed in two dogs to evaluated in vivo viability and biocompatibility of encapsulated islets implanted into the splenic sinus by venouse reflux. Two weeks after auto transplantation, the plasma insulin levels in the splenic vein and artery of two dogs were assayed. In the first dog, serum insulin level was 1 microU/ml both in the vein and the artery and increased, after glucagon (1.0 mg) injection, to levels of 9 microU/ml in the vein, but still kept 1 microU/ml in artery, as well as in the second one. Histological and electron-microscopical examination of the spleen revealed that encapsulated islets remained morphologically intact and the surface of agarose capsules showed no significant adherence of fibroblasts and inflammatory cells. Functional efficacy of the microencapsulated islets was determined using five totally-pancreatectomized diabetic dogs as recipients without immunosuppression. Defined quantity of microencapsulated islets from outbred mongrel donors were grafted through the catheter into omental tissue of the pancreatectomized recipients. All dogs had various degrees of reduced insulin requirements. In three of five recipients, the average fasting glucose values were controlled under 120 mg/dl for 28, 42, 49 days without exogenous insulin, which received totally 4.3 x 10(3), 7.3 x 10(3) and 1.0 x 10(4) (IE/kg) of microencapsulated islets, respectively. In conclusion, the present study indicates that the agarose-based microencapsulated islets can function in large diabetic animals, resulting in the independence of exogenous insulin therapy for prolonged periods without the need for immunosuppression. PMID- 9869851 TI - Chronic rejection. A general overview of histopathology and pathophysiology with emphasis on liver, heart and intestinal allografts. PMID- 9869864 TI - Processing, storage and experimental transplantation of human fetal pancreatic cells. PMID- 9869863 TI - A two-step digestion process and LAP-I cold preservation solution for human islet isolation. AB - Pancreatic islet transplantation has a high potential for treating diabetes mellitus, but long-lasting insulin independence has not been achieved in type I diabetic patients. In order to obtain better results, improvement is needed in many areas. The first area is the islet isolation process. The requirements for an islet isolation method are: 1) to produce a maximum number of healthy islets without demanding a high quality of donor pancreas; 2) to be able to perform the procedure with fewer numbers of personnel who may be without extensive skills and expertise; and 3) to lower isolation costs. In order to achieve these objectives, we have made two important modifications to the isolation process. One is the development of a new preservation solution, LAP-I and the other is the use of a two-step process for pancreas digestion, involving a short warm collagenase digestion, followed by cold mechanical digestion without collagenase. We also use a clear plastic digestion chamber in order to visualize the process. The chamber cover is designed to facilitate frequent removal of digested tissue fragments. The overall procedure is simple and straightforward, requires less manpower and is cost effective. Our procedure is described in detail, and its advantages are discussed. PMID- 9869865 TI - Advantages of microencapsulation as an immunoprotection method in the transplantation of pancreatic islets. PMID- 9869866 TI - Porcine neonatal pancreatic cell clusters (NPCCs): a potential source of tissue for islet transplantation. AB - This is a short review of porcine neonatal pancreatic cell clusters (NPCCs) which might eventually be useful for beta cell replacement therapy in people with diabetes. The current success with islet allograft transplantation is reviewed and is problematic because only partial success has been obtained and the shortage of human islet tissue means that only a small fraction of people with diabetes would be able to benefit. For these reasons there is considerable interest in xenotransplantation, with pigs being a particularly attractive source. The relative merits of early fetal, late fetal, neonatal and adult porcine tissue are discussed. Neonatal tissue has several attractive features, with their hardiness and potential for growth being especially noteworthy. NPCCs are harvested after digested and dispersed clumps of cells are kept in culture for 7 days. The NPCCs consist mainly of duct cells, protodifferentiated cells and mature endocrine cells. The protodifferentiated cells are either double or triple stained for insulin, cytokeratin 7, glucagon, pancreatic polypeptide, or somatostatin. When transplanted into diabetic nude mice it usually takes weeks before glucose levels are normalized, and during that time differentiation and growth of the graft can be observed. Potential strategies for controlling xenograft rejection are mentioned, with these being immunosuppression, induction of tolerance, immunobarrier devices, and gene transfer approaches. PMID- 9869867 TI - Porcine islet xenotransplantation utilizing a vascularized bioartificial pancreas. AB - Successful pancreas/islet transplantation restores normal glucose metabolism in patients with insulin dependent diabetes mellitus (IDDM) but requires chronic immunosuppression which is associated with morbidity and mortality. Immune exclusion devices containing pancreatic islets (bioartificial pancreas) are designed to provide glycemic control without immunosuppression. The immune exclusion is achieved by separating islets from the host by semipermeable membranes. Small molecules such as glucose, insulin and nutrients pass through, whereas immune lymphocytes and immunoglobulins are excluded by the membrane and unable to cause destruction of the islets. Use of xenogeneic islets (i.e., porcine islets) in the device also circumvents the shortage of human donor organs. This report provides a brief summary of our experience with vascularized bioartificial pancreas (VBAP) containing allogeneic and xenogeneic islets for treatment of experimental diabetes in dogs and describes our plans for a clinical trial of the VBAP in patients with IDDM. PMID- 9869868 TI - Tilapia Brockmann Bodies: an inexpensive, simple model for discordant islet xenotransplantation. AB - Brockmann bodies (BBs), large anatomically discrete islet organs found in some teleost fish, are much more easily harvested than pancreatic islets from mammalian donor sources. Tilapia islets, when transplanted into streptozotocin diabetic athymic nude mice, will produce long-term normoglycemia and mammalian like glucose tolerance profiles. Our laboratory has used tilapia BBs as an inexpensive model for studying islet xenograft rejection between discordant species. When transplanted into immunocompetent diabetic mice, tilapia BBs reject in roughly 7-8 days. Results to date suggest that tilapia islets are very immunogenic and that encapsulation is necessary to achieve long-term function in euthymic recipients. Continuous, high-dose immunosuppression using 15-DSG is also effective but causes lethal bone marrow suppression. Less aggressive immunosuppression regimens have achieved only modest prolongation of mean graft survival time. Thus far, immunomodulation techniques and transplantation into immune-privileged sites have been ineffective at prolonging graft survival. Tilapia islets currently represent an excellent, inexpensive donor source for discordant islet xenotransplantation studies. In the not distant future, encapsulated islets harvested from transgenic tilapia bearing humanized tilapia insulin genes may also play a role in establishing clinical islet xenotransplantation as a useful treatment modality for type I diabetes mellitus. PMID- 9869869 TI - Tolerance induction for islet transplantation. AB - Type I diabetes is a systemic autoimmune disease. Although transplantation of pancreatic tissues restores glucose homeostasis, grafts are affected by acute and chronic rejection as well as re-occurrence of autoimmune destruction. One newly recognized promising strategy to interrupt these detrimental processes is hematopoietic chimerism induced by bone marrow transplantation (BMT). The application of hematopoietic chimerism has three domains in the treatment of Type I diabetes mellitus: (1) tolerance induction to pancreas or pancreatic islet grafts; (2) prevention of the re-occurrence of autoimmune processes in the graft; (3) prevention of the onset of overt diabetes once the pre-diabetic state is clearly identified. Unfortunately, conventional BMT is associated with significant morbidity and mortality due to graft-versus-host disease (GVHD), failure of engraftment and lethal conditioning. The risk of these complications cannot be justified in the treatment of non-malignant diseases including Type I diabetes. This chapter will outline potential strategies to achieve hematopoietic chimerism without the risk of deadly complications. With these strategies, it may be possible to apply hematopoietic chimerism in the treatment of Type I diabetes, both to induce tolerance to islet allografts as well as to intervene and interrupt the autoimmune process in its early stages. PMID- 9869870 TI - Acquired systemic tolerance to islet allografts induced by intrathymic inoculation of alloantigens--a brief review. PMID- 9869871 TI - Overview of the problem of cancer in organ transplant recipients. PMID- 9869872 TI - Transmission of cancer from organ donors. AB - One hundred and seventeen of 270 (43%) recipients of organs obtained from donors with malignancies had evidence of transmitted cancers. In 9 instances these were removed from renal allografts immediately prior to transplantation. Including these cases there were 45 recipients of organs in which a neoplasm involved the allograft, 6 others in whom adjacent structures were invaded, and another 66 patients who had distant metastases. Precautions to prevent cancer transmission include meticulous preoperative screening of donors, careful examination of all organs at the time of harvesting, biopsy of any suspicious lesions, and routine donor autopsy, if possible. PMID- 9869873 TI - Evaluation of transplant candidates with pre-existing malignancies. AB - A retrospective study was done of the recurrence rates of 1297 preexisting tumors in renal transplant recipients. Of 1137 neoplasms that were treated prior to transplantation, the recurrence rate was 21%, and it was 33% in 99 cancers treated after transplantation. Fifty-four percent of recurrences in the pretransplant-treated group occurred among malignancies treated within 2 years of transplantation, 33% in those treated 2-5 years before transplantation, and 13% among those treated more than 5 years pretransplantation. Among the 31 neoplasms in the last group 52% of recurrences occurred within 2 years after transplantation. Among those cancers treated pretransplantation the highest recurrence rates occurred with breast carcinomas (23%), symptomatic renal carcinomas (27%), sarcomas (29%), bladder carcinomas (29%), nonmelanoma skin cancers (53%) and multiple myeloma (67%). In the tumors treated posttransplantation 39% of recurrences were from nonmelanoma skin cancers. The bulk of evidence suggests that immunosuppressive therapy facilitates the growth of residual cancers. With some exceptions a minimum waiting period of 2 years between treatment of a neoplasm, with a favorable prognosis, and undertaking renal transplantation is desirable. A waiting period of approximately 5 years is desirable for lymphomas, most carcinomas of the breast, prostate or colon, or for large (> 5 cm) symptomatic renal carcinomas. No waiting period is necessary for incidentally discovered renal carcinomas, in situ carcinomas, and possibly tiny focal neoplasms. As it is highly unlikely that most candidates for nonrenal transplantation can be kept alive for a two year waiting period nonrenal transplantation can be undertaken in patients who have been treated for major cancers, provided that the disease appears to have been adequately controlled, and that the stage of the malignancy does not have a poor prognosis. PMID- 9869874 TI - Liver transplantation for primary and metastatic liver cancers. PMID- 9869875 TI - De novo cancers complicating renal transplantation: experience in the Nordic countries. AB - OBJECTIVES: This report summarizes the experience in the Nordic countries with de novo cancer complicating renal transplantation and the prospects of a new Nordic EU-SPONSORED study (EURO-CAT) covering all kinds of organ transplantations. METHODS: Epidemiological studies are performed using standard methods of cohort analysis. The expected numbers of cancer cases in each stratum are calculated by applying the stratum- and site-specific cancer incidence rate of the national population to the appropriate number of person-years at risk. The observed/expected ratio is taken as the relative risk (standardized incidence ratio, SIR). Differences in SIRs between subcohorts are tested by evaluating the rate ratio. Laboratory methods include immunopathological, virus serological cytokine and tissue type analyses. Treatment includes discontinuation of immunosuppression and long-term high dose aciclovir. RESULTS: In our most recent analyses an incidence 3-5 times higher than that of the general population has been found, including most types of tumors, however with an uneven pattern with emphasis on lip-cervical-, vulvar-, urological-, skin cancers and lymphoprolipherative disorders (PTLD). Most tumors occur early (incl. PTLD) after transplantation, tumors of the skin however later and with an increase with time. In our present analysis we follow all types of organ transplantation, up to the most recent date, to consider the many new immunosuppressive agents for their risk of inducing cancer. Laboratory analyses have pointed to IL-10 as a possible cofactor in the development of EBV-induced PTLD. Treatment with aciclovir and discontinuation of immunosuppression may in some cases lead to recovery from the malignancy, sometimes even without harming the graft function. PMID- 9869876 TI - Skin cancers in organ transplant recipients. AB - Organ transplant recipients on immunosuppressive therapy are prone to skin cancers, especially squamous cell carcinomas developing on sun-exposed areas. Their frequency increases with time after transplantation reaching 40-70% of the patients after 20 years. Squamous cell carcinomas tend to be multiple and may have a life-threatening course. Most studies concern kidney transplant recipients but new data are now available on recipients of other organs. Carcinogenic factors include mainly immunosuppressive treatments, UV light and human papillomaviruses; the role of genetic factors is more equivocal. Melanomas and other rare tumors such as Merkel cell tumors or sarcomas are also increased. Surgical excision with histological examination represents the treatment of choice. When lesions become multiple and/or aggressive, additional therapeutic methods are necessary, such as topical or oral retinoids and in some cases, reduction of the immunosuppressive treatment. Radiotherapy should be reserved to limited cases. Prevention must be undertaken by a regular dermatological examination and sun protection. PMID- 9869877 TI - Clinicopathologic features of posttransplant lymphoproliferative disorders. AB - Posttransplant lymphoproliferative disorders (PTLD) are primarily B lymphocyte tumors which are related to the Epstein-Barr virus. Recent studies have more clearly delineated neoplastic from hyperplastic forms of this disease. Factors associated with increased PTLD risk include recipient EBV seronegative status and heavy immunosuppression. The clinical presentation of PTLD is reviewed and lesser known features such as respiratory compromise or localization of tumors to skin are highlighted. The pathologic classifications of PTLD are surveyed and related to one another. Newer approaches to therapy, including the use of monoclonal antibodies and adoptive cellular immunotherapy are discussed. PMID- 9869878 TI - Treatment and prognosis of post-transplant lymphoproliferative disease. AB - Post-transplant lymphoproliferative diseases (PTLDs) are a clinically and morphologically heterogeneous group of lymphoid proliferations. They represent a life-threatening complication of solid organ transplantation. The mechanisms of their pathogenesis are not yet fully understood. A combination of impaired immunity, oncogenic consequences of immunosuppressive therapy and EBV infection may play a role. Studies on incidence, treatment and prognosis are difficult because of the small number of cases occurring at each transplant center and the lack of reliable classification. Overall mortality remains high even though 25% of patients require no other measure than reduction in immunosuppression which must be the first step of treatment. Several treatments are currently used but more adequate classification as well as multicenter studies are urgently needed because many questions remain with regard to therapeutic strategy. Late-onset monoclonal tumors may be treated by conventional chemotherapy, while EBV-positive PTLDs may benefit from other approaches such as antiviral therapies or immunologic modulation of tumor functions. PMID- 9869879 TI - Kaposi's sarcoma in renal transplant recipients. AB - One hundred and seventeen of 270 (43%) recipients of organs obtained from donors with malignancies had evidence of transmitted cancers. In 9 instances these were removed from renal allografts immediately prior to transplantation. Including these cases there were 45 recipients of organs in which a neoplasm involved the allograft, 6 others in whom adjacent structures were invaded, and another 66 patients who had distant metastases. Precautions to prevent cancer transmission include meticulous preoperative screening of donors, careful examination of all organs at the time of harvesting, biopsy of any suspicious lesions, and routine donor autopsy, if possible. PMID- 9869880 TI - Ano-genital neoplasia in renal transplant patients. AB - Ano-genital neoplasia is about 20 x more common in renal transplant patients than the general population. Neoplasms in the immunosuppressed are more morbid and mortal because: patients are younger; tumors are more undifferentiated; they have more and larger foci; more sites are involved; neoplasms tend to persist, recur and progress; and there are more complications from treatments. Intraepithelial neoplasia engenders some morbidity. Invasion is rarer, but when it occurs, it is always morbid, and all too often mortal. Invasive ano-genital cancer is primarily preventable because the lower genital and anal tracts are accessible to inspection, cytologic screening, endoscopy and biopsy. Prime prevention is avoiding infection with the Human Papilloma Virus (HPV). Next best is detecting HPV/intraepithelial neoplasia early with frequent inspection, cytology and liberal biopsies; and then removing any condylomas or intraepithelial neoplasia that develop. PMID- 9869881 TI - Etiology and pathogenesis of posttransplant tumors: new insights into viral oncogenesis. PMID- 9869882 TI - Societal issues in organ transplantation. PMID- 9869884 TI - Two initiatives designed to maximize the potential for organ donation from intensive care units. PMID- 9869883 TI - Cultural and psychological dimensions of human organ transplantation. AB - Human organ transplantation is practiced in local cultural worlds that shape beliefs about appropriate conduct for its development and application. The psychological response of individuals to the transplant experience mediate and condition its life-changing force in the context of family and community. In this paper, three cases are examined to illustrate the impact of cultural and psychological influences on human organ replacement therapies. First, we explore brain death and its implications for the definition of death and the procurement of organs. A case example from Japan provides the framework for addressing the cultural foundations that contribute to perceptions of personhood and the treatment of the body. Second, we examine marketing incentives for organ donation using a case from India where, until recently, explicit forms of financial incentives have played a role in the development of renal transplantation involving non-related living donors. Third, we focus on the psychological remifications of organ transplantation using a case that demonstrates the profound experience of being the recipient of the "gift of life". Resolution of scientific and ethical challenges in the field of organ transplantation must consider the complex and significant impact of cultural and psychological factors on organ replacement therapies. PMID- 9869885 TI - Guidelines to assist national authorities with regard to the formulation of transplant policies and laws. PMID- 9869886 TI - Organ and tissue donation: are minorities willing to donate? AB - The Task Force on Organ Transplantation (DHHS,1986) addressed the issue of increasing organ donation. The Report of the Task Force recommended that "educational efforts aimed at increasing organ donation among minority populations be developed and implemented, so that the donor population will more closely reflect the ethnicity of potential transplant recipients, in order to gain the advantage of improved donor and recipient immunologic matching (DHHS,1986). Donor rates for minorities has increased as follows: 16% in 1988 to 23% in 1995 among cadaveric donors and 24% in 1988 to 28% in 1995 among living donors. The improvement in donor rates among minorities may positively affect the transplantation success rate experienced by organ recipients of the same race. Strategies must be implemented that will increase the effectiveness and frequency of communication between minority patients and the medical community. An increase in the effectiveness of communication between potential minority donor families and the health care community will contribute to the process of Consciousness Raising as discussed by Prochasksa. The result of increased awareness of the organ donation and transplantation process may have a favorable impact on organ donation. The media has, through public service announcements, paid advertising and entertainment programming, attempted to promote discussion of organ donation in the community and within families. Johnson et al. discussed Mexican-American and Anglo-American Attitudes Toward Organ Donation. The primary impediment contributing to the disparity of consent rates between Mexican-American and Anglo American population occurs with regard to the donation of organs of relatives. Johnson stated that this impediment to organ donation can be effectively addressed by promoting family discussion. Communication within families will inform surviving next of kin of the desire of the deceased to be an organ donor and hence improve the likelihood of the donor family consenting to organ donation (Johnson et al., 1988). The promotion of communication within families must continue to be a goal of the transplant community regardless of race/ethnicity. Despite the efforts of the government, the transplant community, the media and the corporate sector to address the critical shortage of donors in the United States, the reality is that no community has the supply of donor organs suitable to meet the need. PMID- 9869887 TI - The Indian transplantation law: promises and effects. PMID- 9869888 TI - Societal challenges in Latin-America. PMID- 9869889 TI - The Eurotold Project. AB - OBJECTIVES: The European Commission funded EUROTOLD Project sought to examine the legal and ethical implications of living donor organ transplantation within Europe, facilitated by a multi-centre study acquiring data on practices, laws, policies, attitudes and decision-making processes. METHODS: Methods involving primary sources included interviews with clinical staff and past and present organ donors and recipients, and questionnaire surveys of transplant centres, individual clinical staff and legal experts. These strategies were supplemented by an examination of secondary sources such as official reports, transplantation literature, etc. RESULTS: The surveys generated substantial new evidence relating to transplant centre policies and practices, and the attitudes of physicians toward living donation generally and the use of certain specific classes of donor. The latter was facilitated by the use of case scenarios, providing a window upon factors influencing judgments in this sphere. The interview data confirmed earlier findings about donor decision-making. CONCLUSIONS: Responding centres and clinicians displayed a fairly liberal attitude toward living donation but substantial diversity nonetheless exists with regard to living donor transplant volumes between centres and surrounding acceptable waiting times for transplant. Further research is required to identify precisely the reasons underpinning such disparity. PMID- 9869890 TI - Attitudes to xenotransplantation: scientific enthusiasm, assumptions and evidence. AB - The use of xenografts could relieve the chronically inadequate supply of human organs for transplantation, but doubts have been expressed about the general acceptability of transplanting animal organs into human. Some researchers and clinicians have chosen to ignore negative attitudes towards clinical xenotransplantation, assuming that people will automatically embrace this new technology when it becomes available. A review of eight studies of attitudes to xenotransplantation did not reveal overwhelming support for it. Particularly negative views were expressed by acute care nurses. Primates have been the donors of choice in clinical xenotransplantation to date, but their continued use is a highly contentious option; the preferred animal donor is now clearly the pig. Animal farming for xenotransplantation is generally regarded as acceptable since animals provide food for man and are an accepted source of items for human use such as heart valves and insulin. Open debate about xenotransplantation must now take place, and present attitudes may change as a result of this. However, it remains to be seen whether xenografts will be widely accepted and used, and the extent to which the chronic shortage of organs for transplantation will thereby be alleviated. PMID- 9869891 TI - Kaposi's sarcoma. AB - Kaposi's sarcoma is an unusual neoplasm that is seen with increased incidence in transplant recipients. Its occurrence in this group as a tumor is noteworthy in that it may regress spontaneously if immunosuppression is reduced or discontinued. Thus, its response in this setting raises the question of whether it might be best classified as a reversible hyperplasia rather than a true malignancy. In this work we will review recent advances in our understanding of Kaposi's sarcoma, while retaining an historical perspective. PMID- 9869893 TI - Body distribution of syngeneic and allogeneic cells from vascularized bone marrow graft. AB - In the previous studies we showed that vascularized bone marrow graft (VBMtx) in transplanted rat hind limb brings about complete repopulation of syngeneic recipient BM cavities and lymphoid organs within 10 days. Transplantation of an equivalent number of bone marrow cells (BMC) in suspension did not produce repopulation until day 30. In this study we present data on transplantation of allogeneic VBM and compare them with those obtained in a syngeneic combination. In the LEW or BN to LEW combination BM cells were labelled with 51Cr, injected i.v., 24 h later the hind limb was amputated and transplanted to a LEW rat. BM cells emigrated from the transplanted limb to the recipient and distributed in BM cavities and lymphoid tissues. In the LEW to LEW combination the level of radioactivity in recipient tibia was after 24 h 0.85, in spleen 2.43, in mesenteric lymph node 0.52%/g of tissue, whereas in the BN to LEW model it was 0.11, 1.83 and 0.15%, respectively. The calculated numbers of BM cells which populated recipient tissues were 8-10-times lower in the allogeneic compared with syngeneic combination. This was probably due to the nonspecific elimination of some subsets of BM cells (allogeneic BMC cytotoxicity). Administration of anti asialo GMI antiserum to the recipient abrogated the cytotoxic effect. Taken together, major differences in kinetics of seeding and repopulation of BMC from VBMTx were found. Elimination of recipient NK cells with AAGMI antiserum attenuated the nonspecific cytotoxic effect. This protocol allows protection of the grafted BMC and increases the efficacy of the transplant. PMID- 9869892 TI - The effect of cyclosporine on regulators of fibrinolysis in plasma from renal allograft recipients. AB - Fibrinolytic disturbances are thought to play an important role in processes leading to deterioration of renal allograft function. We investigated the effect of CsA therapy on the regulation of fibrinolysis in kidney graft recipients by measuring plasma concentration and activity of plasminogen activators (tPA, uPA) and their inhibitors (PAI-1, 2). We found an increase in tPA activity and in PAI 1 concentration as well as a decrease in PAI-1 activity in renal allograft recipients as compared to healthy controls, but did not confirm a correlation between these observations and CsA administration. tPA and PAI-2 concentrations as well as uPA activity did not significantly differ between the studied groups. We showed a significant decrease in uPA plasma concentration in patients treated with azathioprine. The significance of this finding is unknown. PMID- 9869894 TI - Microchimerism following allogeneic vascularized bone marrow transplantation--its possible role in induction of posttransplantation tolerance. AB - We have noticed that bone marrow transplanted in a vascularized limb graft providing a continuous supply of donor BMC may prolong the survival time of skin graft from the same donor. The question arises whether the raised microchimerism plays a role in the prolonged survival of skin allograft. The aim of the study was to follow the development of microchimerism after allogeneic vascularized bone marrow transplantation (VBMTx) concomitantly with the rejection processes of transplanted skin. The BN rats served as donors and LEW rats as recipients of VBMTx and free skin flap allograft. Hind limb was transplanted followed by a full thickness skin graft on the dorsum. Cellular microchimerism was investigated in recipients of VBMTx and skin grafts in blood, spleen, mesenteric lymph node and bone marrow with monoclonal antibody OX27 directed against MHC class I polymorthic RTI on BN cells and quantitatively analysed in FACStar. In VBMTx group free skin flap survived 70 days after weaning of CsA. Intravenous infusion of BMC in suspension equivalent to that grafted in hind limb did not prolong skin graft survival after cessation of CsA therapy. Donor-derived cells could be detected in VBMTx recipients as long 70 days after wearing of CsA but not in recipients of i.v. suspension BMC grafting. PMID- 9869895 TI - Protection of heart and rejection of lymphocyte allografts from the same donor in recipients of donor-specific transfusions. AB - Organ allografts survive in hosts treated with immunosuppressive drugs. The question arises as to whether cells isolated from organ or tissue of an allogeneic donor and transplanted to a genetically disparate recipient can also benefit from the immunosuppressive regimen. We reported previously that the DST (donor specific transfusion) recipients accept heart allografts but reject hyperacutely i.v. infused lymphocytes from the same as DST donor. The present study was devoted to elucidation of the mechanism of these divergent processes. Syngeneic BN hearts and mesenteric lymph node lymphocytes were transplanted to LEW rats pretreated one week previously with donor specific blood transfusions. The allogeneic BN lymphocytes transplanted i.v. to LEW rats receiving one BN DST were rejected hyperacutely within 6 hrs, whereas BN heart grafts transplanted to the BN DST-treated LEW recipients survived 14 +/- 2 days. Adoptive transfer of LEW anti-BN DST sera to naive LEW rats caused destruction of the transplanted BN lymphocytes. The LEW BN DST recipients possessed IgG and IgM class alloantibodies binding to BN lymphocytes and heart endothelial cells. mAbs against MHC class I (OX18) and class II (OX6) antigens neither blocked binding of antibodies of DST recipient sera to BN lymphocytes nor protected the preincubated BN lymphocytes against destruction after transplantation. Western blot analysis revealed that alloantibodies from DST-recipient sera bound strongly to BN lymphocyte membrane proteins of 60 kd m.w. but not to 45 kd and 30 kd MHC class I and II proteins. Taken together, DST has no protective effect on intravenously transplanted cells. In contrast, it accelerates the rejection. Alloantibodies present in DST recipient sera "shield" antigens on the surface of organ allograft endothelial cells, thereby protecting them from recognition and cytotoxic effect. Simultaneously, these alloantibodies "opsonize" the intravenously transplanted lymphocytes and facilitate their halting in lymphoid organs and subsequent lysis. Antibodies other than those directed against MHC seem to mediate both these processes. The results of these studies provide also evidence that the effector mechanism of rejection may be different depending on location of the graft, in the lymphoid as in case of transplanted lymphocytes or in non-lymphoid tissues as heart grafts. PMID- 9869896 TI - Cyclosporin A decreases lymphocyte migration to the heart allograft through suppression of their L-selectin expression. AB - Cyclosporin A (CsA) changes the distribution of the circulating pool of lymphocytes and decreases their traffic to organ allograft. The mechanism of this process is complex and includes, among others, inhibition of induction of nuclear factor of activated T cells and suppression of GM CSF and E-selectin expression. We studied the expression adhesion molecules CD11a, CD18, CD44, CD54 and CD62L on the thoracic duct lymphocytes of rats treated with CsA. The 7-day administration of CsA evidently decreased the expression of CD62L but did not affect the other adhesion molecules. Lower concentration of CD62L molecules on the surface of circulating lymphocytes may influence their migration to allograft and distribution in host lymphoid tissues. PMID- 9869898 TI - Psychological rules of communication with relatives of a potential organ donor- the Polish experience. AB - Increase of number of organs for transplantation depends on the positive attitude of the general public toward cadaveric organ donation and transplantation. This attitude is shaped as the result of education performed by mass media and as a result of individual experience of various people with the health-care service. Whenever a decision on post-mortem donation of a deceased is unknown, there are his close relations who might express his will. The aim of this article is to draw attention to several problems of psychological nature, which refer to proceeding with relatives of organ donors. A course and result of talks with relatives of a potential organ donor depend on a number of factors, which are pointed out below. Within the frames of this interaction, basic rules of effective communication are presented, including message on death of a close person in case of brain stem death diagnosis. Furthermore, examples are presented of the most frequent errors of those who talk with relatives, scope of appearing difficulties and several methods to avoid and/or overcome occurring problems. PMID- 9869897 TI - Mitochondrial activity after cold preservation of pancreatic islet cells treated with pefloxacin (PFX). AB - Mitochondrial energetic and oxidative dysfunctions caused by free radical production trigger release of proinflammatory cytokines involved in organ rejection. The aim of this study was to investigate the role of a fluoroquinolone drug, pefloxacin (PFX) and those of various cold preservation solutions on pancreatic beta cell viability. Our data clearly demonstrate that islet cell viability, as determined by glucose-stimulated insulin secretion, is directly correlated with reduced expression of microsomal cytochrome P-450IIIA. Moreover, IL-2, a known mediator of apoptosis was found to be downregulated, whereas TNF alpha had been upregulated for the first 18 hours after pefloxacin administration. These results demonstrate that pefloxacin downregulates the expression of cytochrome P-450IIIA isozyme and regulates the production of TNF alpha and IL-2. Thus, we postulate that the presence of pefloxacin in the pancreatic islet cells before organ preservation facilitates increased cell viability. PMID- 9869899 TI - Acute failure of the transplanted kidney--pathophysiology, diagnosis and prevention. AB - Acute failure of the transplanted kidney is a major problem in the early posttransplant phase and is recognized as a major cause of graft loss. Early renal transplant dysfunction is mainly due to ischemic damage (acute tubular necrosis), rejection, infection, and cyclosporin A toxicity. Less common causes include bleeding, ureteral obstruction, urinary leak, venous thrombosis, and stenosis or occlusion of the renal transplant artery. Recent advances in both invasive (renal biopsy) and non-invasive (imaging and biochemical) techniques have improved specificity and sensitivity of the diagnosis of the acute renal failure. Several procedures which aim to prevent the kidney transplant failure have recently been introduced. Although they were shown to reduce the incidence of early allograft failure, their influence on the long-term graft survival remains to be proven. PMID- 9869900 TI - Technical aspects of orthotopic liver transplantation. PMID- 9869901 TI - Kinetics of in vitro immune responses of T and B cells during tolerance induction by sirolimus. AB - OBJECTIVES: The purpose of the study presented herein was to examine immune performances of rat heart allograft recipients immunosuppressed with sirolimus (SRL, rapamycin; Rapamune, Wyeth-Ayerst, Princeton, NJ). METHODS: The immune performances of lymphocytes harvested from SRL-treated Wistar Furth (WF; RT1u) recipients of Buffalo (BUF; RT1b) heart allografts were examined on days 7, 14, and 90 postgrafting. RESULTS: Whether derived from normal WF rats, SRL-treated WF heart recipients, or SRL-untreated WF heart recipients, pan-T cell population purified from the lymph nodes or spleens on day 7 or 14 displayed similar responses to phytohemaglutinin, anti-T cell receptor R73 monoclonal antibody, donor-type BUF, or third-party Brown Norway alloantigenic stimulators. There was no in vitro evidence of suppressor T cells in SRL-treated recipients. The frequencies of anti-BUF-specific cytotoxic T cells, as shown by limiting dilution analysis, were similar in the short- (days 7 or 14) and in the long- (day 90) term surviving recipients. SRL treatment did not affect the expression of interleukin-2 (IL-2) messenger RNA (mRNA) by T helper 1 (Th1) or of IL-4 and IL 10 mRNA by Th2 cells on days 7 and 14 postgrafting, but did induce selective activation of Th2 cells on day 60 postgrafting. Administration of SRL induced the production of non-complement (C')-fixing IgG2c BUF-specific alloantibodies that appeared in the sera of unresponsive recipients on day 14 postgrafting and reached a peak concentration on day 120 postgrafting. In contrast to untreated recipients that rejected BUF heart allografts, all SRL-treated WF recipients failed to produce C'-fixing BUF-specific alloantibodies. CONCLUSIONS: SRL promotes long-term selective activation of Th2 cells and the production of non-C' fixing IgG2c blocking antibodies. PMID- 9869902 TI - Predictor of transplanted kidney deterioration following pregnancy--daily urine protein loss or serum creatinine concentration? AB - Increasing number of female kidney recipients deciding to born a baby makes us to investigate the impact of pregnancy on graft survival. The aim of study was to find the parameter that would have the predictive value for graft function following delivery. Scr and P in 33 pregnant renal allograft recipients (mean age 27.1 +/- 6.1 yrs) treated with pred + aza + CsA were studied for 6 mo before, during and 6 mo following delivery. As measured only by Scr graft function was stable in all pts (1.4 +/- 0.05 mg/dl). Significant rise in Scr following pregnancy was found in 6 of 33 pts. This "unstable" group was compared with 27 patients with "stable" despite pregnancy graft function. Proteinuria, but not Scr differentiated groups prior to pregnancy. The estimation of P prior to conception seems to be more potent parameter to predict kidney graft deterioration following pregnancy than Scr alone. Increase in P during and following pregnancy in "unstable" pts may reflect the acceleration of subclinical (not yet manifested with rise of Scr) chronic graft rejection due to pregnancy-induced hiperfiltration. PMID- 9869903 TI - Measurement of radial bone mineral density in patients after heart transplantation. AB - Limited physical activity, steroidotherapy and immunosuppression are known risk factors for the development of osteoporosis. The purpose of our current work was to investigate whether patients after heart transplantation (Htx) have an increased incidence of osteoporosis. We compared bone mineral density (BMD) in 32 post-transplant patients with a reference group of 1548 healthy age-matched males. Measurement of BMD was carried out with a Dtx 100 Osteometer on the distal and ultradistal segment of the non-dominant radius. Our results revealed a decreased BMD in HTx patients ranging from 6.9 to 10% in the ultradistal (p = 0.0446) and from 0.4 to 3.5% in the distal segment (p = 0.0593). PMID- 9869904 TI - Comparison of the different strategies for cryopreserving and storage of the bone marrow CD34+ cells. Possibility of unprogrammed rate freezing and storage at -80 degrees C mechanical freezer. AB - We have compared the efficiency of the different strategies allowing for a long term storage of a human CD34+ bone marrow cells. Accordingly, the aliquots of CD34+ cells isolated from bone marrow were frozen using: controlled rate freezing equipment, or freezer unprogrammed in a -80 degrees C mechanical freezer. After freezing, CD34+ cells were subsequently stored for one month in a liquid nitrogen tank at -196 degrees C or in mechanical freezer at -80 degrees C. We have found that both the viability and the recovery of clonogeneic progenitors of CD34+ cells samples stored at different temperature were similar. Therefore, regarding the costs and simplicity, we recommend the unprogrammed freezing and storage of human CD34+ cells at -80 degrees C in a mechanical freezer as a convenient, inexpensive, and reliable method for storing marrow for transplantation. This data also indirectly indicate that the aliquots of the CD34+ cells can be shipped frozen on dry ice (-80 degrees C), and that these cells will maintain viability under this conditions. Furthermore, in this study we have confirmed validity of our earlier observation that human CFU-Meg progenitors are more sensitive to cryopreservation. PMID- 9869905 TI - Legislative aspects of organ and tissue donation in Belgium. PMID- 9869907 TI - The revival of transplantation. PMID- 9869906 TI - Organ transplantation in Hungary. PMID- 9869908 TI - New transplant legislation in Sweden. The Transplantation Act and its introduction. PMID- 9869909 TI - Organizational aspects of organ procurement. PMID- 9869911 TI - Pregnancy and delivery after liver transplantation. AB - The first case of pregnancy in a patient after an orthotopic liver transplantation (OLT) in Poland is presented. A 21-year-old woman was liver grafted 3 years prior to the pregnancy. Before having conceived the patient's graft function was stable. The woman was on immunosuppressive therapy with cyclosporine A and prednisolone. During pregnancy no significant changes in biochemical tests of liver function and liver blood flow were noted. Starting at the second trimester, a slight anemia occurred and the quantity of blood platelets continued to decrease, the latter having been observed immediately after the transplantation. The intrauterine growth of the fetus was monitored by ultrasound and the assessment of blood flow to the placenta was made. No abnormality was observed. In the second and third trimester the presence of HCV RNA in the serum was found. In the 41-st week of pregnancy labor commenced. The threat of intrauterine infection indicated a cesarean delivery. The newborn weighed 4180 g and had an Apgar score of 10. The cesarean section, as well as puerperium, was normal. The immunosuppressive therapy was continued, and antibiotics were administered for prophylactic reasons. During the first month the infant was treated with antibiotics because of pneumonia and the suspicion of meningitis. Nine months after the delivery, the patient's health is satisfactory and the baby is making normal progress. PMID- 9869912 TI - Organ transplantation in Austria. PMID- 9869910 TI - Successful full-term pregnancy in a patient three and a half years after a heart transplant. AB - The patient is a 28 year old woman who received a heart transplant in 1992 secondary to hypertrophic cardiomyopathy with unremarkable post-operative course. In the period immediately post transplantation the patient was on a four-drug immunosuppressive regimen which was subsequently changed to standard three-agent therapy. This therapy was continued until the patient became pregnant. In the first trimester only Cyclosporine (CsA) was used, and thereafter, the patient was continued on the previous three agent regimen. Toward the end of pregnancy a rise in systolic pressure was observed, but the child was delivered by spontaneous vaginal delivery without complications in the 38th week of pregnancy. The newborn weighed 3320 g and was in good health. A sharp fall in the newborn CsA blood levels was observed post delivery reaching zero level on the third day of life. At the present time, both mother and baby are in good health, 6 weeks after delivery. PMID- 9869913 TI - Inflammatory mediators, cells and their products in acute host alloresponsiveness. PMID- 9869914 TI - Extracellular matrix proteins: bystanders or active participants in the allograft rejection cascade? AB - The role extracellular matrix (ECM) proteins may play in the host immune cascade triggered by organ transplantation is largely unknown. We have employed well defined rat cardiac allograft model to test the hypothesis that the expression of ECM components, such as fibronectin (FN) or laminin (LN), represents an integral part of the host rejection response in transplant recipients. Indeed, treatment of rats with anti-LN Ab profoundly affected lymphocyte recirculation pathways, and selectively decreased deposition of alloreactive lymphocytes at the graft site and in host peripheral lymph nodes. Moreover, cardiac rejection was accompanied by markedly increased intragraft expression of both LN and FN. The infiltrating mononuclear cells, however, accumulated selectively in FN-rather than LN-rich cardiac areas. We also observed distinct temporal expression and spatial distribution patterns of FN variants in rat cardiac allografts and isografts. The local synthesis of FN, in both allografts and isografts was increased as early as at 3 h post-transplant. The expression of FN in cardiac allografts but not in isografts, remained high during later intervals to the point of ultimate rejection. The newly synthesized FNs derived from graft infiltrating macrophages and arterioles, and included EIIIA +, EIIIB +, and CS-I + splicing variants. The EIIIA was preferentially enriched in the earlier, whereas EIIIB and CS-I were selectively enhanced in the later, rejection phases. Finally, treatment of rat recipients with synthetic FN-CS-I peptides prevented the incidence of early (acute) and late (chronic) rejection, supporting the notion that distinct FNs may play a fundamental role in the host immune cascade triggered by organ transplantation. These data offer potential novel sites for intervention in the control of transplant rejection, and contribute to the development of refined therapeutic strategies based upon new concepts of host immunosuppression. PMID- 9869915 TI - Sirolimus, a potent new immunosuppressive drug for organ transplantation. PMID- 9869917 TI - Renal transplantation--the Norwegian model. PMID- 9869916 TI - Nucleotide sequences of three H-2K and three H-2D complementary DNA clones coding mouse class I MHC heavy chain proteins. AB - OBJECTIVES: The polymerase chain reaction (PCR)-based method was used to obtain and sequence three H-2K and three H-2D mouse complementary DNAs (cDNA) of class I major histocompatibility complex (MHC) molecules. METHODS: Messenger RNA was isolated from Conconavalin A-activated splenocytes of C57BL/10 (H-2b), C3H (H 2k), and Balb/c (H-2d) mice. We designed H-2K- and H-2D-specific primers as well as a common downstream primer based on previously published mouse class I MHC sequences. Using the PCR method and selective primers we isolated and sequenced H 2Kb and H-2Db cDNAs of C57BL/10, H-2Kk and H-2k cDNAs of C3H, as well as H-2Kd and H-2Dd cDNAs of Balb/c strains. RESULTS: Analysis of the nucleotide sequences documented similarity between our three H-2K cDNA sequences and all mouse MHC class I sequences available in the GenBank. Similarly, our three H-2D sequences were homologous with all mouse class I MHC sequences deposited in the GenBank. Our H-2K and H-2D sequences were also identical to numerous published sequences. CONCLUSIONS: Using these mouse cDNAs, we plan to determine the localization of polymorphic in vivo immunogenic amino acids in class I MHC H-2K and H-2D alloantigens. PMID- 9869918 TI - Social, legal and medical limitations of organ transplantation in Poland. PMID- 9869919 TI - Organ transplantation in Austria. PMID- 9869920 TI - The National Transplant Organization donation evolution and transplant activity in Spain. PMID- 9869921 TI - Syngeneic and allogeneic blood transfusions produce hematopoietic and immune effects. AB - Blood transfusions (BT) should be considered as transplantation of blood elements. They evoke various alterations in the immune responsiveness of blood recipients, contributing to an increased risk of infection and cancer recurrence. The exact mechanism by which blood transfusions induce a state of reduced immune responsiveness remains unclear. Relatively little is also known about immune changes occurring in lymphoid compartments other than blood following blood transfusions. In the present study, the effect of syngeneic and allogeneic blood transfusions on hemopoiesis and immune responsiveness was examined in a rat model. Transfusions of both syngeneic and allogeneic blood caused an increase in the bone marrow myeloid and lymphoid lineage cell compartments as well as a rise in the percentage of OX7+ stem cells in bone marrow. It was followed by a release of OX7+ stem cells into blood circulation, higher in the case of rats transfused with syngeneic blood. The changes in hemopoiesis were accompanied by a diminished responsiveness of blood, spleen and bone marrow lymphocytes to mitogens in both groups of rats. Data point to early changes in distribution and reactivity of bone marrow and lymphoid cells following blood transfusion. PMID- 9869922 TI - Clinical course of concomitant Hbv and Hcv infection in renal allograft recipients. AB - We evaluated the impact of concomitant infection with Hepatitis B virus (HBV) and Hepatitis C virus (HCV) on the clinical course after renal transplantation (Tx). In 335 patients (pts) transplanted between 1991 and 1993 we found 30 (9%) recipients who were positive for Hepatitis B surface antigen (HBsAg) (ELISA, Organon) and anti-HCV antibodies (immunoblot assay Lia Tek) preTx. Chronic liver disease (CLD) (two-fold or greater increase in serum ALT and AST levels for at least six months) developed in 40.7% coinfected pts as compared to 24.4% and 25.7% pts infected only with HCV or HBV, respectively. Maintenance immunosuppression consisted of P + Aza + CsA, mean follow-up time was 28 +/- 15 months. The mean time of the onset of CLD was 3.0 months (range: 1-18 months) after Tx. Percutaneous liver biopsy performed in 5 CLD pts revealed chronic active hepatitis (CAH) in 4 and chronic persistent hepatitis (CPH) in 1 pt. Four pts who had CAH and were positive for HCV RNA (RT PCR) in serum and for HBcAg in liver tissue, received interferon-alpha therapy for 6 months. Clinical improvement of liver function was observed in all of them, but none cleared HBsAg or HCV RNA. One pt lost his graft due to acute rejection. Concomitant infection with HBV and HCV is associated with the high risk of development of CLD early after Tx. We recommend that pretransplant evaluation of both anti-HCV and HBsAg positive pts should include liver biopsy to exclude potential recipients with CAH. PMID- 9869923 TI - Diagnosis and monitoring of cytomegalovirus infection after liver transplantation in children. AB - Cytomegalovirus infection (CMV) complicated the posttransplant course in 9 of 24 children after liver transplantation. We found specific antibodies (IgG and IgM) to be of very low value in diagnosis and monitoring of CMV infection after liver transplantation. Detection of CMV-DNA by PCR method in the blood or urine was very useful for diagnosis, but less for monitoring of the course of disease and its treatment. Measurements of early immediate CMV antigen (IEA), in peripheral blood leucocytes allowed for very early diagnosis of CMV infection and correlated well with the course of disease and response to treatment of the patient. PMID- 9869924 TI - The effect of nephrectomy on the outcome of renal transplantation in patients with polycystic kidney disease. AB - Although various forms of polycystic kidney disease (PKD) account for up to 10% of patients requiring renal replacement therapy and severe complications may arise from these kidneys, no clear indications for pretransplant nephrectomy have been defined so far. A total of 104 renal transplants in three pediatric and 96 adult patients suffering from PKD were analysed retrospectively with regard to patient and graft survival in relation to pretransplant or posttransplant nephrectomy and no nephrectomy. Of these 99 patients, 25 had had either unilateral (19) or bilateral (6) nephrectomy sometime before transplantation and 10 patients between 3 and 81 months after transplantation. All patients received Cyclosporine-based immunosuppression. One-year patient and graft survivals for recipients of a first cadaveric renal graft (n = 91) were 94% and 92%, for recipients of second or third graft (n = 13) 89% and 78%. One- and five-year patient survival rates for patients with or without pretransplant nephrectomy were 100% and 100% vs 92% and 84%, respectively. One- and five-year graft survival rates were 100% and 93% for pretransplant nephrectomy patients vs 89% and 74% for the non-nephrectomy group (p < 0.05). Patients not undergoing nephrectomy sometime after transplantation had the same patient but better five year graft survival when compared to the posttransplant nephrectomy group (89% vs 52%). In patients with early posttransplant urinary tract infection, which is considered in this analysis as a cyst-related complication, graft survival at one year was 77% but 97% in patients without this complication. From these data it is recommended that polycystic kidneys should be removed before transplantation if cyst-related complications occur repeatedly. Posttransplant nephrectomy can be performed with no mortality and should be carried out whenever clinically indicated. PMID- 9869925 TI - Acute rejection and delayed graft function--risk factors of graft loss. AB - Acute graft rejection and delayed function are considered to be the major risk factors of short-term as well as long-term graft survival. We studied the impact of these factors on graft outcome among 109 renal transplant recipients. All recipients were treated with triple drug protocol. The recipients were divided into two groups: I group included 57 patients with delayed graft function (DGF), II group included 52 patients with immediate graft function (IGF). We studied graft survival, incidence of acute rejection, serum creatinine levels and the cause of graft loss for patients in both groups. Acute rejection episodes occurred in 49% of patients from DGF group and 45% of patients from IGF group. Graft survival in IGF group was better than in DGF group. Actuarial graft survival at 1, 2, 3 and 4 years in examined groups was 84%, 82%, 72%, 65% vs. 92%, 86%, 84%, 84%, respectively. One-year graft survival in patients with acute rejection from DGF group and IGF group was significantly lower than in patients who remained rejection free (69%, 74% vs. 94%, 96%). We concluded that delayed graft function decreases long-term graft survival, while immediate graft function has an excellent impact on graft outcome. Acute graft rejection is the strongest risk factor of graft loss. PMID- 9869926 TI - Effect of Sandimmun Neoral on the level of bivalent cations in erythrocytes of kidney transplant recipients. AB - In a 4 month study, a group of 16 patients with stable renal graft function receiving triple immunosuppressive therapy including cyclosporin A (Cy A) were investigated for the levels of calcium, magnesium and zinc in erythrocytes. The patients were randomized to be converted to the new microemulsion formulation (Sandimmun Neoral) in a 1:1 fashion (n = 8) or to continue with the classical formulation (Sandimmun) (n = 8). The concentrations of creatinine, phosphate, alkaline phosphatase activity, calcium, magnesium and zinc were measured twice a month in blood plasma. The concentration of calcium, magnesium and zinc in erythrocytes was also measured. The concentration of magnesium in blood plasma and erythrocytes during the study showed no deviation from normal values. The level of zinc in erythrocytes was almost twice as high as in normal healthy controls and was not dependent on Cy A formulation. Calcium content in erythrocytes of patients receiving Sandimmun was 27.6% higher than in healthy persons. Conversion of the patients to Sandimmun Neoral normalized the calcium concentration in erythrocytes and caused a transient increase of calcium levels in blood plasma. PMID- 9869927 TI - The effect of endocrine disturbances on hemodynamic stability of brain dead organ donors. I. Thyroid function. PMID- 9869928 TI - Comparison of intravascular ultrasound and quantitative angiography for evaluation of coronary artery disease in the transplanted heart. PMID- 9869929 TI - History of organ transplantation in Warsaw. A personal perspective. PMID- 9869930 TI - Organ transplantation in Poland. A registry report. AB - Solid organ transplantation program in Poland is very limited. The main reason of this is organ shortage. All the organs procured are transplanted. It is a general feeling that recently issued legal regulations will favorably influence organ donation, however the improvement may be expected within a few years and will require very active educational campaign among society and medical staff. National Transplant Council prepared the plans of organ transplantation for the nearest five years. These plans were met with full support of the Ministry of Health and Welfare and Health Committee of the Polish Parlament. PMID- 9869931 TI - History of bone marrow transplantation in Poland. PMID- 9869932 TI - Factors limiting renal transplantation program in Poland. AB - The shortage of donor organs has been the main obstacle to the expansion of transplantation programs. Recent public opinion survey documented acceptance of the cadaveric kidney procurement in our country but some reluctance to brain death and presumed consent concepts. Recently, the survey was carried out within the medical community to find out whether the level of knowledge and the attitude toward donation has an influence on the slow development of the transplantation program. A questionnaire was addressed to: 1010 general practitioners, neurosurgeons and anesthesiologists; 926 ICU and neurosurgical nurses and 1760 students of 12 medical schools of the country. Vast majority of doctors, nurses and last year medical students accept retrieval and transplantation of kidneys and the heart, but not of the liver. Acceptance of this procedure among junior medical students and university students was lower (78% vs 98%). Most of the respondents would agree to donate their kidneys and other organs, but 20% would protest against harvesting of the organs from their relatives. 100% of the physicians and 80% of medical students and nurses accept the brain death concept(which is accepted only by 60% of non medical university students) but only 44% of the doctors are prepared to switch off the respirator after diagnosis of brain death if harvesting is not taking place. Only half of the physicians would notify the transplantation unit about the possibility of organ retrieval. The reasons mentioned for such decision included fear of negative judgment of the local community and problems with deceased relatives. 60% of physicians talking to the family about retrieval would ask for the relatives' consent despite the fact, that the transplantation law in Poland is based on the presumed consent of each individual. The results of the knowledge survey among medical students documented inadequate medical education concerning problems of transplantation. Educational campaign is needed to promote and extend the cadaveric organ transplantation in our country. PMID- 9869933 TI - Function of endocrine organs in kidney transplant patients. AB - Several factors are involved in the persistence of endocrine alterations after renal transplantation, among which the following are to be mentioned: 1) duration of chronic uraemia before renal transplantation; 2) residual function of the patients' native kidneys; 3) quality of function of the renal graft; 4) modulation of secretion, transport, and degradation of hormones, and/or 5) altered target organ responsiveness to hormones induced by immunosuppressive drugs (glucocorticoids, azathioprine, cyclosporin A) or altered internal environment. In kidney transplant patients the following endocrine abnormalities are to be mentioned: dissociation of the physiological relationship between aldosterone synthesis and function of the renin-angiotensin system, abnormal volumetric regulation of arginine vasopressin secretion, suppressed responsiveness of cortisol secretion to stimulatory manoeuvres, persistent secondary hyperparathyroidism, relative deficiency of insulin (induced by glucocorticoid therapy), with consequent carbohydrate intolerance or even diabetes mellitus, suppressed response of gastrin and pancreatic hormone secretion to a test meal, and reduced responsiveness of atrial natriuretic peptide secretion to central hypervolaemia. Episodes of acute graft rejection are characterized by endocrine alterations similar to those seen in patients with acute or chronic renal failure. PMID- 9869934 TI - Erythropoietin production after kidney transplantation. AB - We studied production of erythropoietin (EPO) in long-term renal allograft recipients with posttransplant erythrocytosis (PTE). Among 951 recipients we found 74 patients with persistent elevation of hematocrit (Htc) value (female > 50%, male > 55%). However, only 63.5% of them had increased red-cell mass ( = "true" erythrocytosis). In all recipients with PTE known causes of secondary erythrocytosis were not found. EPO titer in peripheral blood was significantly higher in recipients with PTE (median 13.5 mIU/mL, range: 0.1-71.5 mIU/mL) as compared to healthy blood donors (median 5.75 mIU/mL, range: 0.1-19.5 mIU/mL) but not different from the group of renal allograft recipients without PTE (median 13.0, range 0.1-71.7 mIU/mL). However, EPO level measured in pretransplant sera was significantly higher in patients who developed PTE (median 16.4 mIU/mL, range: 1.0-281.2 mIU/mL) than in recipients without PTE (median 8.3, range: 1.0 50.3 mIU/mL). A significant difference in EPO level between systemic and effluent blood from native kidneys was found in 6 out of 14 recipients with PTE who underwent catheterization. After phlebotomy patients with PTE responded with higher increase in peak EPO titer than healthy blood donors (527 +/- 473% versus 194.5 +/- 44.2%, p < 0.05). Our results suggest that PTE develops spontaneously due to increased EPO production. Despite elevated EPO levels, regulation of EPO release remains preserved. PMID- 9869935 TI - Graft rejection in sensitized recipients. AB - Recipient sensitization to MHC antigens is among the most critical of problems currently facing clinical transplantation in terms of magnitude and impact. Approximately 20% of all patients awaiting kidney transplantation have elevated levels of broadly reactive alloantibodies, resulting from multiple transfusions, prior failed allografts, or pregnancy. These highly sensitized patients experience an increased rate of graft rejection, compared to unsensitized individuals, which is often irreversible and difficult to control by currently used immunosuppressive agents. The evidence suggests that the common denominator in the pathogenesis of allograft rejection in presensitized recipients is the combined cellular and humoral immune alloreactivity. Although the relative significance of these responses varies, all successful immunosuppressive therapies have common three elements of diminished or abrogated mononuclear cell activation, cytokine expression, and endothelial cell activation. The main target for the antibody-mediated damage, the vascular endothelium, may also be affected by nonspecific effector mechanisms, complement, coagulation, phagocytic cells, and their products. The dissection and better appreciation of the complexity of the rejection cascade may be important for the much needed progress in the management of allograft rejection in sensitized hosts, and may provide insights relevant to the development of xenotransplantation. PMID- 9869936 TI - Intraglomerular fibronectin and laminin turn-over in chronically rejected kidney allografts in humans. AB - Chronic rejection is primarily responsible for the late loss of allografted organs and remains an important clinical problem. Chronic rejection in the kidney is characterised by arteriolosclerosis and nephrosclerosis, glomerulonephritis and interstitial fibrosis. Recently, a large number of studies have indicated that proteolytic enzymes play important roles as mediators of glomerular injury. The aim of the study was to assess intraglomerular fibronectin and laminin contents as well as cysteine proteinases in activity chronically rejected human kidneys. We investigated kidney tissue from graftectomy specimens obtained from 11 patients with end-stage renal disease following chronic rejection. A group of 9 patients undergoing nephrectomy because of cancer served as a control group, but only not involved parts of the kidneys were used. When intraglomerular laminin contents were related to DNA content, significant accumulation in chronically rejected allografts was found in comparison to controls (382 +/- 171 micrograms per microgram DNA and 190 +/- 82 micrograms per microgram DNA, respectively, p < 0.01. The accumulation of fibronectin was higher than in controls, however the difference was not significant. When proteinase activity was related to intraglomerular DNA content, significantly enhanced cathepsin B and L activity was found in rejected kidney allografts (57 +/- 16 nmol AMC/min per mg DNA) in comparison to controls (15 +/- 2 nmol AMC/min per mg DNA). Summarizing, we observed accumulation of fibronectin and laminin in glomeruli and simultaneously an excess of proteolytic activity in human chronically rejected kidneys. The above phenomenon indicates that a very active metabolic process takes place in glomeruli during rejection. PMID- 9869937 TI - Extended donor criteria--a possible way to enhance the number of potential heart donors. PMID- 9869938 TI - Cardiac transplantation in Krakow. PMID- 9869939 TI - Cultured parathyroid cells allotransplantation without immunosuppression for treatment of intractable hypoparathyroidism. AB - Cultured, viable and functioning ABO compatible parathyroid cells were allografted in 18 nonimmunosuppressed patients with a postoperative hypoparathyroidism. Variable, but promising biochemical and clinical results were obtained. Clinical and biochemical observations have documented graft function up to 14 months. The mechanism of cessation of function of implanted cultured cell suspension remains unknown. Some parameters, suggest a rejection mechanism but other mechanisms can not be excluded. This suggests that some form of immune modulation may be necessary to improve further PT allograft survival in recipients off immunosuppressive therapy. PMID- 9869941 TI - The effect of clinical and biochemical donor parameters on pancreatic islet isolation yield from cadaveric organ donors. AB - The recovery of pancreatic islet cells from cadaveric donors for allotransplantation may depend on the functional condition of the pancreas of the donor prior to organ harvesting. We examined donor hemodynamic and biochemical parameters and their effects on the subsequent yield of islet cells after harvesting. All pancreata were flushed and preserved in University of Wisconsin (UW) solution and digested using automated method within 8 hours. In the first analysis, digestions were divided into high-yield (> 2000 IEQ/g pancreas), and low-yield (< 2000 IEQ/g) groups and donor variables were averaged for each group. Donors whose pancreata yielded > 2000 IEQ/g received significantly greater amounts of Dopamine (14.43 micrograms/kg/min vs 9.35 mg/kg/min, p = 0.05). The daily urine output between groups was also significantly different. Maximum systolic blood pressure (SBP), minimum systolic blood pressure, use of vasopressin, length of hospitalization, and maximum base deficit were compared between the two groups. Less severe hypoglycemia (lowest blood glucose 143 mg/dL vs 107 mg/dL, p = 0.02) and lower amylase levels (36.2 U vs 80.7, p = 0.07 were noted in the high-yield group. A trend towards higher islet yields was associated with lowest hourly urine output > 60 (2040 IEQ/g vs 1649 IEQ/g p = 0.09), maximum SBP > 200 (2097 vs 1673, p = 0.07, and surprisingly, lowest SBP below 80 (2013 vs 1742, p < 0.1). Amount of fluids administered prior to procurement had no influence on islet yield. In conclusion, hemodynamic variables such as urine output, systolic blood pressure, and degree of pressor support were modestly associated with successful islet isolation. The preliminary data suggest that better multifactor donor analysis is imperative for standardization and monitoring of multiorgan donors. The association of higher blood glucose levels with successful isolation may also be related to resuscitation with dextrose containing fluids. PMID- 9869940 TI - Experience with pancreas islets separation, immunoisolation and cryopreservation. AB - Experience of Warsaw Pancreas Laboratory is presented. Some improvement in the methods of rat, human and pig pancreases digestion, and in identification of Langerhans islets by means of intravenous injection of I-DTZ was achieved. For immunoisolation of islets, 2 methods were elaborated: capsules containing alginate/polyethyleneimine/protamine/heparin membrane prepared by modified Sun method, and microcapsules based on Zekorn method. Biocompability of hollow fibers, prepared with polypropylene (PP), surface modified PP (PPS) and polysulphone (PS) was assessed in vitro. Only PS fibers were fully compatible. It was shown, that the mixture of exocrine tissue did not influence in vitro insulin secretion, providing that alginate in which islets are embedded remain gelled. The efficacy of 3 methods of islets cryopreservation was compared: freezing in "semicontrolable" conditions, in programmable Kriomedpol machine, and vitrification. The highest percentage of frozen/thawed living cells, and the most reliable results were obtained with Kriomedpol method. PMID- 9869942 TI - Hematopoetic progenitor cells transplantation in Poland: structure and clinical practice. AB - The present activity of BM/PBPC transplantations is based on government funded transplantation centers. About 50 alloBMT/PBSC or 1.3 transplant per million per year were performed in the last two years with a tendency to further increase. An increase in the number of autoBMT/PBSC is also seen with about 75 autotransplantations or 2 transplantations per million of people per year. Indications and standard laboratory and clinical practices in Poland are similar to those in other European countries. However, chemotherapy prevails over total body irradiation (TBI) in conditioning regiments. An average long term overall survival of patients with hematological malignancies after alloBMT is higher in paediatric transplantations than in adults (0.56 vs 0.34, p < 0.03) and in patients with a standard risk than in cases with advanced disease (0.48 vs 0.36). PMID- 9869943 TI - Reconstitution of lymphoid tissue after vascularized bone marrow transplantation. AB - We reported previously that vascularized bone marrow transplantation (VBMT) in an orthotopic hind limb graft brings about complete repopulation of bone marrow cavities in lethally irradiated syngeneic recipients within 10 days. Intravenous infusion of an equivalent volume of bone marrow cell suspension was evidently less effective. The purpose of this study was to investigate the reconstitution of immunocompetent compartments of lethally irradiated syngeneic rats after VBMT. Lewis rat hind limbs were transplanted orthotopically into irradiated recipients. Ten days after irradiation and bone marrow transplantation, bone marrow, mesenteric lymph nodes and sera from rats were harvested. Responsiveness of mesenteric lymph node lymphocytes (MLNL) to mitogens and lymphocyte proliferation in the presence of sera and bone marrow cell (BMC) culture supernatants was measured. Our studies have shown that vascularized bone marrow transplantation brings about rapid replenishment of lymphoid organs of lethally irradiated syngeneic recipients. The repopulating subsets were fully responsive to mitogens. Sera from reconstituting rats had no evident effect on proliferation of mature lymphocytes. Intravenous infusion of BMC in suspension, in a number equivalent to that grafted in hind limb transplant, was less efficient in reconstituting lymphoid tissue. PMID- 9869944 TI - Bone banking for transplantation in orthopaedic reconstructions. AB - Results of orthopaedic reconstructions in 1014 patients with the use of preserved allogenic bone grafts are presented. The method of grafts preservation seems to be of significance in surgical applications; deep-freezing is more favorable in more comparison with lyophilization. The result of treatment after bone transplantation is dependent on numerous biological and clinical variables. PMID- 9869945 TI - Intracoronary arterial occlusion: a novel technique potentially useful for ablation of cardiac arrhythmias. AB - To develop a new technique for ablating arrhythmias by interrupting coronary perfusion of the myocardium, we studied six mongrel dogs, weighing 20-35 kg. Under angiographic guidance a microcatheter (1.0 mm diameter) was introduced into a branch of the left anterior descending or posterior descending coronary artery. A detachable platinum coil (0.0254 cm diameter, 3 cm length) soldered onto a stainless-steel delivery wire (Guglielmi) was inserted through the microcatheter and advanced to occlude the arterial branch. A 0.5-mA electric current applied to the proximal end of the delivery wire resulted in intravascular thrombosis due to attraction of the negatively charged blood cells, platelets, and fibrinogen to the positively charged platinum coil. In approximately 4.5 minutes, as the thrombus was formed, electric current dissolved the soldering and detached the platinum coil from the delivery wire. Electrocardiograms showed focal ST-T changes but no ventricular tachyarrhythmias. Pathologic studies revealed thrombosis around the platinum coil and well-demarcated focal ischemia/infarction that was correlated with elevation of cardiac enzymes. We conclude that intracoronary arterial embolization and electrothrombosis using an electrolytic platinum coil can be selectively performed in a very small coronary arterial branch, resulting in a limited area of myocardial damage. This technique is potentially useful for ablating arrhythmias and may be safer and more controllable than intracoronary alcohol infusion. PMID- 9869946 TI - Ultrarapid train stimulation versus conventional programmed electrical stimulation for induction of ventricular arrhythmias in patients with coronary artery disease. AB - Conventional programmed electrical stimulation (PES) of the ventricle is useful for establishing inducibility or noninducibility of clinical ventricular arrhythmias (VA) but is complex and time consuming. The present study was designed to compare a standard PES protocol with an alternative method using ultrarapid train stimulation in patients with VA and coronary artery disease (CAD). A prospective, randomized, crossover design was used. During each session in the electrophysiology laboratory, patients were studied using both the trains and PES protocols in randomized order. In 82 matched pairs of comparisons in 50 patients, results were concordant in 85% (p < 0.0001). There were no differences related to type of clinical arrhythmia or to the presence of antiarrhythmic drugs. There were no significant differences in the induction of nonclinical arrhythmias with the two methods (p < 0.0001 for concordance). There were no significant differences related to the cycle length of the trains (10, 20, or 30 ms, equivalent to 100, 50, or 33 Hz). The number of drive-extrastimuli sequences and the time required to complete the trains protocol was significantly shorter (p < 0.0001) using trains versus PES. Ultrarapid train stimulation provides results in CAD patients that are comparable with those of conventional PES protocols. There is a significant savings in time, adding practical value to intrinsic electrophysiologic interest. Trains may be useful when multiple inductions are desirable, for example, in the setting of antitachycardia pacing parameters in an implantable defibrillator (ICD), during ICD implantation, or in other circumstances where the main question is inducibility of ventricular arrhythmias. PMID- 9869947 TI - Divergence of endocardial QT interval components during programmed electrical stimulation including observations during induction of sustained ventricular tachyarrhythmias. AB - Measurements were made in 12 normal subjects and during induction of sustained ventricular tachyarrhythmias in 31 patients with remote myocardial infarction. QT interval measurements were made semiautomatically with computer assistance and the total QT interval was divided into early (QT1) and late (QT2) components. QT intervals and QT interval dispersion between two right ventricular endocardial sites were plotted against the degree of prematurity of the last extrastimulus (S2, S3, or S4). In the control group, total QT and QT1 intervals shortened with increasing prematurity of the last extrastimulus (p < 0.001). Slopes (positive) were steeper with faster pacing rates (600, 500, or 400 ms) and more extrastimuli (1 to 3). The relationship between QT2 intervals and prematurity of the last extrastimulus was flat, but the slope was slightly negative (p = 0.05 to < 0.001) and did not vary with changes in pacing cycle length or number of extrastimuli. QT interval dispersion in the control group was minor (95% CI 0-40 ms). During induction of sustained ventricular tachyarrhythmias, total QT and QT1 intervals were longer (y intercepts) than in the control group (p < 0.05 at 400-ms pacing cycle length) and their dispersion was increased (p < 0.05). Generally, QT2 intervals were shorter (p < 0.05 at 600-ms pacing cycle length) during induction of ventricular arrhythmias in comparison with the control group but dispersion was increased (p < 0.05 at 400-ms pacing cycle length). QT intervals and QT interval dispersion show an orderly and predictable relationship with prematurity of the last extrastimulus in normal subjects. These patterns differ during induction of sustained ventricular tachyarrhythmias. Such differences may be exploited to derive clinically predictive and useful measurements. PMID- 9869949 TI - Adenosine and atrial fibrillation. PMID- 9869948 TI - Dose and rate-dependent effects of adenosine on atrial action potential duration in humans. AB - Adenosine provokes atrial fibrillation (AF) in some patients with paroxysmal supraventricular tachycardia (PSVT). Which patients are more susceptible to develop atrial fibrillation after the administration of adenosine to terminate PSVT is unknown. We prospectively measured atrial action potential duration (APD) at incremental doses of 3, 6, and 12 mg of adenosine at paced cycle lengths (CLs) of 600, 500, and 400 ms in 25 patients. Bolus injection of adenosine decreased APD at 90% repolarization in a dose- and rate-dependent manner. During paced CLs of 600, 500, and 400 ms, decreases of 8%, 13%, and 19% (p < 0.05), respectively, were found after bolus administration of 3 mg of adenosine. After 6 mg of adenosine, the APD shortened by 12%, 19%, (p < 0.05), and 27% (p < 0.01), respectively. After 12 mg of adenosine, the APD shortened by 15%, 27% (p < 0.05), and 38% (p < 0.01), respectively. Transient AF occurred in 4 of 25 (16%) patients, all during paced CLs of 400 ms, and after adenosine 6 mg in one patient and 12 mg in three patients. Adenosine shortens atrial action potential duration in a dose- and rate-dependent manner. Whether patients with faster rates during PSVT and those given higher doses of adenosine are more prone to develop atrial fibrillation remains to be determined. PMID- 9869950 TI - Arrhythmogenic right ventricular dysplasia: clinical results with implantable cardioverter defibrillators. AB - Arrhythmogenic right ventricular dysplasia is a clinical entity characterized by fatty infiltration of the right ventricle and left bundle morphology ventricular tachycardia occurring in young patients. The most common cause of death is tachyarrhythmic. Pharmacological and nonpharmacological therapies, including implantable cardioverter defibrillators, have been used to treat the arrhythmias. However, right ventricular endocardial leads in this population may be associated with an increased risk of perforation and suboptimal sensing and defibrillation efficacy due to the diseased right ventricle. We report on 12 patients with arrhythmogenic right ventricular dysplasia who were treated with implantable cardioverter defibrillators. The mean age was 31 +/- 9 years (range 15-48). Patients presented with presyncope (5), syncope (4), or cardiac arrest (3). All patients had electrocardiographic abnormalities characteristic of the condition. During programmed electrical stimulation nine patients had sustained ventricular tachycardia, while three patients had no inducible arrhythmia. Transvenous leads were placed in nine patients. In these patients pacing thresholds were significantly higher, R-wave amplitudes were significantly lower and defibrillation thresholds were not significantly different than in a cohort of patients without right ventricular dysplasia. There were no acute or chronic complications of right ventricular lead placement. Follow-up averaged 22 +/- 13 months (range 1-45). There was one sudden death at 1 month of follow-up. Of the 12 patients, 8 have had appropriate therapy delivered by the implantable defibrillator. Six patients are currently on sotalol to reduce the frequency of implantable defibrillator discharges. In conclusion, implantable cardioverter defibrillators with nonthoracotomy leads are feasible and safe in patients with arrhythmogenic right ventricular dysplasia. The frequency of appropriate therapy is high, supporting the use of implantable cardioverter defibrillators in this population. PMID- 9869951 TI - Evaluation of a programming algorithm for the third tachycardia zone in a fourth generation implantable cardioverter-defibrillator. AB - The clinical efficacy of electrical algorithms for termination of slow ventricular tachycardia (VT) and ventricular fibrillation (VF) in implantable cardioverter-defibrillators (ICDs) is well established. Such algorithms have not been equally well defined for fast VT reversion. We report the testing of a prospectively designed algorithm for ICDs to treat fast VT that is inherently less responsive to antitachycardia pacing than slow VT. Fourth-generation ICD devices were programmed to three prospectively defined tachycardia detection zones as follows: cycle lengths < or = 260 ms for VF, 270-330 ms for fast VT, and > 330 ms for slow VT. The initial selected therapy for the VF zone was a high energy biphasic shock (> 15 J), while a 3- or 5-J biphasic shock was usually administered for fast VT, and antitachycardia pacing was initially attempted for slow VT. Initial therapy was followed by backup therapy with high-energy shocks. Twenty-eight patients, 24 of whom were males, all with organic heart disease, with a mean age of 65 +/- 9 years, received either a Medtronic 7219D (23 patients), 7219C (2 patients), 7218SP1 (2 patients), or 7218C (1 patient) ICD with a nonthoracotomy lead system. The defibrillation threshold was 10 +/- 5 J. At predischarge electrophysiologic testing, a single 3- or 5-J shock terminated all episodes of fast VT tested. During a follow-up of 18 +/- 9 months, there were four nonarrhythmic deaths. Fourteen patients (50%) had a total of 21 VF, 44 fast VT, and 202 slow VT episodes. Twenty-three of 24 (96%) VF, 33 of 39 (84%) fast VT, and 193 of 202 (95.5%) slow VT episodes were terminated with the first delivered therapy in each therapy algorithm (p = NS). The overall efficacy of the entire electrical therapy algorithm was 100% for VF, 100% for fast VT, and 98% for slow VT episodes (p = NS). No patient experienced syncope or presyncope during fast VT or VF in this study. We conclude that a third detection and therapy zone can be successfully programmed in ICDs to define a range of fast VT episodes that can be effectively terminated with lower energy cardioversion shocks with comparable success and freedom from arrhythmic symptoms to electrical therapies used for slow VT and VF. PMID- 9869952 TI - Pulsing microwave energy: a method to create more uniform myocardial temperature gradients. AB - Microwave energy has been proposed as a possible technique to create large myocardial lesions. Achieving a uniform myocardial temperature gradient during microwave ablation may prevent excessive endocardial temperatures while maintaining temperatures at depth. The goal of the current study was to examine the ability of microwave (MW) pulsing to achieve a more uniform myocardial temperature gradient. Using an in-vitro ovine endocardial model, we measured tissue temperature at 0.5-mm, 2.0-mm, and 3.5-mm depths in a circulating saline bath. MW energy was delivered at 20 W at 915 MHz for 30 seconds. Pulse configurations of 1 second on-1 second off, 3 seconds on-3 seconds off, and 5 seconds on-5 seconds off, with 30 seconds of total MW time were compared with 30 seconds continuous. Maximum temperatures at 0.5 mm were significantly lower at 63.2 +/- 5.89 degrees C for the 1-second pulse compared with 83.5 +/- 7.31 degrees C for the continuous-energy delivery. Pulse configurations 3 seconds on-3 seconds off and 5 seconds on-5 seconds off also resulted in a significantly lower surface temperature than continuous-energy delivery. However, temperature at the 2.0-mm and 3.5-mm depth created by the pulsing delivery were similar to those achieved during continuous-energy delivery. Thus, microwave pulsing achieves a lower endocardial temperature and results in a more uniform temperature gradient. These techniques may prevent the excessive endocardial damage that may result in an increased risk of thrombus formation and embolization. PMID- 9869953 TI - Catheter ablation of ventricular tachycardia associated with remote myocardial infarction: utility of the atrial transseptal approach. PMID- 9869954 TI - Sustained bundle branch reentry in a patient with hypertrophic cardiomyopathy and nondilated left ventricle. AB - Ventricular tachycardia is a well-known complication in patients with hypertrophic cardiomyopathy. We report the case of a patient with hypertrophic cardiomyopathy with easily inducible monomorphic ventricular tachycardia. Electrophysiology study demonstrated that bundle branch reentry was the mechanism of the tachycardia. The tachycardia was rendered non-inducible by radiofrequency ablation of the right bundle branch. PMID- 9869955 TI - History. The specific conduction system in the left ventricle of the human heart. PMID- 9869956 TI - On MADIT and future clinical trials for prevention of sudden cardiac death. Multicenter Automatic Defibrillator Implantation Trial. PMID- 9869957 TI - Defibrillation efficacy with endocardial electrodes is influenced by reductions in cardiac preload. AB - Little is known about the effects of cardiac preload and cardiac geometry on defibrillation efficacy with endocardial electrodes. We studied nine pigs implanted with an endocardial lead system in the normal and reduced preload state. In the reduced preload state, a balloon catheter was inflated in the inferior vena cava (IVC) for 20 seconds prior to the induction of ventricular fibrillation (VF). Complete occlusion of the IVC and reductions in preload were confirmed by observing deformation of the contrast-filled balloon, a reduction in cardiac size by fluoroscopy, and reductions in ventricular pressures. Biphasic shocks were delivered after 10 seconds of VF using a recursive up-down protocol. VF was induced 20 times for each preload state, and the 50% effective doses (ED50) for energy, current, and voltage were estimated by averaging all shocks for that state. At reduced preloads, energy decreased from 12.1 +/- 3.0 J (+/- SD) to 10.5 +/- 2.9 J (p < 0.01), voltage decreased from 415 +/- 51 V to 390 +/- 51 V (p < 0.05), and current decreased from 8.6 +/- 1.5 A to 7.6 +/- 1.5 A (p < 0.01), while impedance rose from 49.2 +/- 3.8 omega to 52.8 +/- 4.4 omega (p < 0.001). We conclude that reducing cardiac preload and cardiac size significantly lowers ED50 defibrillation energy, current, and voltage. This outcome may be caused directly by the decrease in blood volume as evidenced by increased impedance and/or may be due to changes in heart geometry and stretch. PMID- 9869959 TI - Effects of high-frequency atrial pacing in atypical atrial flutter and atrial fibrillation. AB - Atypical atrial flutter has, hitherto, been relatively refractory to termination by rapid atrial pacing. High-frequency pacing (HFP) in the atrium, for termination of atrial flutter or atrial fibrillation (AF), and the electrophysiologic effects related to it have not been examined. We examined the clinical efficacy, safety, and electrophysiologic mechanisms of HFP using 50-Hz bursts at 10 mA applied at the high right atrium in patients with atypical atrial flutter (group 1) or AF (group 2), using a prospective randomized study protocol. Four burst durations (500, 1000, 2000, and 4000 ms) were applied at the high right atrium repetitively in random sequence in 22 patients with spontaneous atrial flutter or AF. Local and distant right and left atrial electrogram recordings were analyzed during and after HFP. HFP resulted in local and distant right and left atrial electrogram acceleration in 8 of 10 patients (80%) in group 1 but caused less frequent local atrial electrogram acceleration (6 of 12 patients) and no distant atrial electrogram effects in group 2 (p < .05 versus group 1). The HFP protocol was effective in arrhythmia termination in 6 of 10 patients in group 1 but in no patient in group 2 (p < .05 versus group 1). Standard HFP protocol applied at the high right atrium can frequently alter atrial activation in both atria and can terminate atypical atrial flutter. Efficacy in AF is limited, probably due to limited electrophysiologic actions beyond the local pacing site. PMID- 9869958 TI - Cardiac pacing in patients with dilated cardiomyopathy. AB - Studies of both the acute and chronic effects of permanent cardiac pacing in patients with dilated cardiomyopathy have produced contradictory results. Similarly, theoretically promising novel pacing techniques such as multisite, His bundle, and right ventricular outflow tract pacing have also yielded mixed results in preliminary studies. In general, at the present time, pacing therapy should be considered investigational for patients with dilated cardiomyopathy who do not have traditional bradycardia indications; nevertheless, pacing therapy may be useful in a small population of patients with dilated cardiomyopathy who have inappropriate timing of left ventricular and left atrial contraction. The beneficial effects of cardiac pacing in some patients with DCM cannot be dismissed out of hand; they must be explained. What is needed is clarification of the best methods to measure any benefits of this therapy: among the options are echocardiographically determined parameters of forward flow and AV valve function, exercise test parameters such as duration and O2 consumption, global and regional ejection fraction, functional classification, quality of life questionnaires, neurohumoral parameters such as plasma atrial natriuretic peptide and catecholamines, and, finally, disease progression and survival. PMID- 9869960 TI - Transvenous cardioversion of atrial fibrillation using low-energy shocks. AB - Recent reports have suggested that transvenous cardioversion of atrial fibrillation is feasible using low-energy shocks and a right atrium coronary sinus electrode configuration. We evaluated in a prospective study the efficacy and safety of low-energy internal cardioversion of atrial fibrillation in 104 consecutive patients. Sixty-two patients presented with chronic atrial fibrillation (group I), 16 had paroxysmal atrial fibrillation (group II), and 26 had an induced atrial fibrillation episode (group III). The mean duration of the presenting episode of atrial fibrillation was 9 +/- 19 months for group I, 4 +/- 2 days for group II, and 18 +/- 7 minutes for group III. Atrial defibrillation was performed using two intracardiac catheters: one was placed in the right atrium (cathode) and the other in the coronary sinus or in the left branch of the pulmonary artery (anode). The catheters were connected to a customized external defibrillator capable of delivering 3/3-ms biphasic waveform shocks with a voltage programmable between 10 and 400 volts. The shocks were synchronized to the R wave. Sinus rhythm was restored in 44 of the 62 patients in group I (70%), in 12 of the 16 patients in group II (75%), and in 20 of 26 patients in group III (77%). The mean voltage and energy required for cardioversion were respectively 300 +/- 68 V and 3.5 +/- 1.5 J, for group I, 245 +/- 72 V and 2.0 +/- 0.9 J for group II, and 270 +/- 67 V and 2.6 +/- 1.2 J for group III. The leading-edge voltage required for sinus rhythm restoration was significantly higher (p < 0.05) in the chronic atrial fibrillation group than in the paroxysmal or induced groups. No proarrhythmic effects occurred for the delivered 686 R-wave synchronized shocks. This study of a large group of patients confirms and extends the results of previous reports. Such findings may have clinical implications for elective cardioversion of atrial fibrillation and the development of an implantable atrial defibrillator. PMID- 9869961 TI - Pacing threshold increase in nonthoracotomy implantable defibrillator leads: implications for battery longevity and margin of safety. AB - Just as a stable defibrillation threshold is required for implantable defibrillators to maintain efficacy and a margin of safety for the conversion of life-threatening ventricular arrhythmias, a stable pacing threshold is also required to provide bradycardia support and pacing to terminate ventricular tachycardias. This article reports the temporal course of pacing thresholds in patients treated with a tripolar, tined endocardial defibrillator lead capable of bipolar sensing and pacing, and defibrillation. Seventeen patients who underwent implantation of an implantable defibrillator system using an integrated bipolar pacing/sensing system were prospectively studied over 18 months. There were 16 males and one female, with a mean age of 69 +/- 5 years (range 61-75 years). At implantation, predischarge, and every 2 months thereafter, the pacing pulse-width threshold was tested at both 2.5 and 5.0 V stimulus amplitudes. After a mean follow-up of 363 +/- 173 days (range 34-597 days), the pacing threshold increased from 0.08 +/- 0.08 ms to 0.5 +/- 0.3 ms at the 2.5 V amplitude (p < or = 0.01, CI 0.57 to -0.27) and from 0.04 +/- 0.02 ms to 0.25 +/- 0.14 ms at the 5.0 V amplitude (p < or = 0.01, CI -0.28 to -0.14). Eight of the 17 patients (47%) received spontaneous implantable defibrillator shocks for clinically detected arrhythmias, and the total number of joules delivered via the leads did not correlate with the pacing threshold changes. We conclude that the pacing threshold for the nonthoracotomy implantable defibrillator lead system studied is not stable and increases with time. This finding has implications for defibrillator battery life in patients who use implantable defibrillators for bradycardia pacing. PMID- 9869962 TI - Early postoperative rise in defibrillation threshold associated with hematoma formation with unipolar defibrillation system. PMID- 9869963 TI - Combined radiofrequency ablation-cooling catheter for reversible cryothermal mapping and ablation. AB - Reversible cryothermal mapping of cardiac arrhythmias has been performed intraoperatively. However, a steerable cooling catheter for reversible mapping has not yet been developed. We therefore developed and tested a cooling system consisting of a -15 degrees C hypertonic saline reservoir and a 7F steerable catheter also capable of radiofrequency (RF) ablation. Using excised ovine hearts placed in a 37 degrees C circulating saline bath, we measured the temperatures at depths of 0 mm, 1 mm, and 2 mm. The temperature after 90 seconds of cooling was 16.5 +/- 2.1 degrees C at 0 mm compared to 23.9 +/- 4.1 degrees C at 1 mm and 31.1 +/- 3.9 degrees C at 2 mm depth (p < 0.01). These data suggest that a 7F steerable combined RF ablation-cooling catheter may achieve temperatures suitable for mapping arrhythmias such as atrial tachycardias and right ventricular outflow tract tachycardias. Further enhancements to achieve lower temperatures at depth may be needed to reversibly map other arrhythmias such as left ventricular tachycardias. PMID- 9869964 TI - Alterations in heart rate following radiofrequency ablation in the treatment of reentrant supraventricular arrhythmias: relation to alterations in autonomic tone. AB - To determine the relation between the creation of endocardial lesions and alterations in autonomic tone, we analyzed heart rate variability in patients undergoing radiofrequency catheter ablation for symptomatic supraventricular tachycardia. Elevated heart rates are frequently noted after radiofrequency catheter ablation for supraventricular arrhythmias. It has been postulated that this elevation may be secondary to alterations in cardiac autonomic tone. Since heart rate variability is a measure of autonomic nervous system activity, we used this technique to examine the heart rate elevation and to characterize postablation autonomic changes. Thirty-eight patients undergoing 44 radiofrequency catheter ablation procedures were included in the study. Total arrhythmic substrates treated included 34 accessory pathways and 13 AV nodes with dual physiology. Twenty-four hour ambulatory electrocardiographic recordings were obtained in a drug-free state prior to, ablation early postablation, and late postablation. Spectral and nonspectral analyses of heart rate variability were performed. Subgroup analyses were also done on specific cohorts. Subgroups included patients undergoing accessory pathway ablations, AV node modifications, and ablation of septal and nonseptal targets. To determine whether the amount of tissue damage was related to changes in heart rate variability, we analyzed the relation between the total energy delivered to the endocardium and the peak change in creatine kinase and heart rate variability. In this population, a significant transient increase in heart rate was noted following radiofrequency ablation. All time and frequency domain parameters of heart rate variability showed significant reversible decreases. These changes were independent of target site and arrhythmia substrate. There was no correlation noted between the changes in heart rate variability and either the total amount of energy applied to the endocardium or the change in creatine kinase. Increased heart rates and decreased heart rate variability occur following radiofrequency catheter ablation for supraventricular tachycardia. Clinically, the predominant effect is that of decreased parasympathetic tone. Since these transient changes are independent of arrhythmic substrate or ablation site in the atria, a rich parasympathetic innervation of the heart is proposed. PMID- 9869965 TI - Bradycardia-induced polymorphic ventricular tachycardia after radiofrequency catheter modification of atrioventricular junction. AB - We report the case of a 59-year-old man with a dilated nonischemic cardiomyopathy who had chronic rapid atrial fibrillation despite several therapies. Radiofrequency modification of the atrioventricular (AV) junction slowed the mean ventricular rate from 120 beats per minute (bpm) to 60 bpm. Five hours after the procedure and during the following 1 week, despite ventricular pacing at 90 bpm, the patient developed nonsustained or sustained polymorphic ventricular tachycardias. Finally, pacing at 90 bpm was successfully used in this patient. In conclusion, patients who have undergone modification of AV conduction may be at high risk of life-threatening ventricular arrhythmias in the first week following the procedure. PMID- 9869966 TI - Polymorphic ventricular tachycardia complicating radiofrequency catheter modification of the AV node for control of rapid ventricular response in atrial fibrillation. PMID- 9869967 TI - Successful ablation of a nonreentrant dual atrioventricular nodal tachycardia. AB - Selective radiofrequency catheter ablation of the slow pathway of the AV node has become the treatment of choice for AV nodal reentrant tachycardia. We describe a case of a nonreentrant AV nodal tachycardia and its successful treatment by slow pathway ablation. PMID- 9869968 TI - Definitions and endpoints for device clinical trials in atrial fibrillation--a pressing need. PMID- 9869970 TI - Transesophageal echocardiograpic imaging of de novo thrombus formation after successful external electrical cardioversion of atrial fibrillation. PMID- 9869969 TI - Complete AV block following mediastinal radiation therapy: electrocardiographic and pathologic correlation and review of the world literature. AB - The clinical, features, serial electrocardiograms, and autopsy findings of a patient with symptomatic complete AV block, who had received mediastinal radiation therapy 8 1/2 years previously, are presented. The cardiac histopathology disclosed immense fibrosis of the conduction system and of the atria and ventricles. The enormous amount of fibrosis was similar in location and intensity to that observed in our previously reported patient (Cohen et al., Arch Intern Med 1981; 141:676-679) who had undergone mediastinal radiation. We conclude that the severe fibrosis was primarily due to radiation, rather than secondary to atherosclerotic coronary artery disease, which also has been described as a consequence of mediastinal radiotherapy. This patient's serial electrocardiograms disclosed evidence of complete block both in the AV nodal area and infra His system, which correlated well with the histopathology. The characteristic clinical features of patients with symptomatic complete AV block post mediastinal radiation therapy are presented, along with a review of the world literature. PMID- 9869971 TI - Pacing mode and long-term survival in elderly patients with congestive heart failure: 1980-1985. AB - Dual-chamber pacing may improve short-term hemodynamics and functional class in some patients with congestive heart failure, even in the absence of conventional indications for pacemaker implantation. However, the impact of different pacing modes on survival of patients with congestive heart failure is controversial. In this retrospective study we analyzed survival data from 546 elderly patients, aged 70 years and older, who underwent implantation of a permanent dual-chamber (DDD, n = 62, DVI, n = 102) or single-chamber (VVI) pacemaker (n = 382) between 1980 and 1985. Survival was further analyzed according to the presence of absence of congestive heart failure, and pacemaker mode (DDD vs. DVI vs. VVI). Overall, dual-chamber pacing (DDD and DVI) was associated with a more favorable long-term outcome when compared with single-chamber ventricular pacing, although differences were only significant for DDD pacing (P = 0.002). When patients with and without preexisting congestive heart failure were analyzed separately, survival following dual-chamber pacing (DDD and DVI) was significantly better than survival following single-chamber pacing in patients without congestive heart failure (P = 0.03), but not in patients with preexisting heart failure (P = 0.139). When patients were analyzed according to the electrophysiological indication for pacemaker implantation, overall survival of patients with AV block (P = 0.0025) but not sinus node dysfunction (P = 0.346) was improved with dual chamber pacing. This survival advantage in patients with AV block following dual chamber pacing was lost in the presence of heart failure (P = 0.11). These findings suggest that dual-chamber pacing, in particular DDD pacing, improves the survival in elderly patients without preexisting congestive heart failure. In contrast to the short-term hemodynamic improvement observed in selected patients with congestive heart failure, dual-chamber pacing in elderly patients with congestive heart failure, paced for conventional indications, is not associated with improved survival when compared with single-chamber ventricular pacing. PMID- 9869972 TI - Early out-of-hospital experience with an impedance-compensating low-energy biphasic waveform automatic external defibrillator. AB - Impedance-compensating low-energy biphasic truncated exponential (BTE) waveforms are effective in transthoracic defibrillation of short-duration ventricular fibrillation (VF). However, the BTE waveform has not been examined in out-of hospital cardiac arrest (OHCA) with patients in prolonged VF often associated with myocardial ischemia. The objective of this study was to evaluate the BTE waveform automatic external defibrillator (AED) in the out-of-hospital setting with long-duration VF. AEDs incorporating a 150-J BTE waveform were placed in 12 police squad cars and 4 paramedic-staffed advanced life support ambulances. AEDs were applied to arrested patients by first-arriving personnel, whether police or paramedics. Data were obtained from PC Data Cards within the AED. Defibrillation was defined as at least transient termination of VF. Ten patients, 64 +/- 14 years, were treated for VF with BTE shocks. Another 8 patients were in nonshockable rhythms and the AEDs, appropriately, did not advise a shock. Five of the 10 VF arrests were witnessed with a 911 call-to-shock time of 6.6 +/- 1.7 minutes. VF detection and defibrillation occurred in all 10 patients. Spontaneous circulation was restored in 3 of 5 witnessed arrest patients and 1 survived to discharge home. Fifty-one VF episodes were converted with 62 shocks. Presenting VF amplitude and rate were 0.43 +/- 0.22 (0.13-0.86) mV and 232 +/- 62 (122-353) beats/min, respectively, and defibrillation was achieved with the first shock in 7 of 10 patients. Including transient conversions, defibrillation occurred in 42 of 51 VF episodes (82%) with one BTE shock. Shock impedance was 85 +/- 10 (39 138) ohms. Delivered energy and peak voltage were 152 +/- 2 J and 1754 +/- 4 V, respectively. The average number of shocks per VF episode was 1.2 +/- 0.5 (1-3). More than one shock was needed in only 9 episodes; none required > 3 shocks to defibrillate. Impedance-compensating low-energy BTE waveforms terminated VF in OHCA patients with a conversion rate exceeding that of higher energy monophasic waveforms. VF was terminated in all patients, including those with high impedance. PMID- 9869974 TI - Implantable cardioverter-defibrillator therapy: influence of left ventricular function on long-term results. AB - The degree of left ventricular impairment in an acknowledged important prognostic marker of long-term outcome for patients being evaluated for implantation of cardioverter-defibrillators. Just how left ventricular function impacts freedom from all-cause mortality, as well as from sudden death and cardiac death, is a subject of current major debate, and is analyzed hereunder from a large, recent multicenter ICD patient cohort. The multicenter database consists of data from 361 patients receiving implantable cardioverter-defibrillators for standard indications, that is, documented episodes of ventricular fibrillation or sustained ventricular tachycardias with poor hemodynamic toleration. Data were collected from 1988 to 1995 at three centers in Germany. Two-hundred and three patients (56%) had a left ventricular ejection fraction (LVEF) > 0.30 (group I), and 158 patients (44%) had a LVEF < or = 0.30 respectively (group II). The mean follow-up was 23.9 months (range 3-98 months). Overall survival at 5 years for group II patients was lower, as expected, at 74.1% versus 94.2%, respectively (P < 0.0001). Mortality was higher for each different cause of death in group II patients than in Group I: sudden arrhythmic deaths, 5 versus 1 (P < 0.048); nonsudden cardiac deaths, 16 versus 5 (P < 0.002); noncardiac deaths, 7 versus 2 (P < 0.03). Group II patients received a higher rate of at least one presumably appropriate shock at 86 (54.4%) versus 89 (43.8%) in group I (P < 0.05). However (and somewhat surprisingly), neither the time from ICD implantation to death, comparing only the patients who died, nor the event-free probability of appropriate shocks due to very rapid, sustained ventricular arrhythmias (> 230 beats/min), including a presumed risk of sudden arrhythmogenic death, differed between groups I and II. Sudden cardiac death was only marginally affected by LVEF (group I, 1.5% actuarial, 5-year survival 99.5%; group II, 3.1% and 95.8%, respectively). Therefore, the lower overall survival in ICD patients with LVEF < or = 0.30 resulted mainly from causes of death that cannot be directly influenced by cardioverter-defibrillator therapy. However, because group II patients had a far higher incidence of at least one ventricular tachyarrhythmia terminated by ICD shocks than group I patients, they also probably derived benefit from ICD therapy. PMID- 9869975 TI - Comparison of three different automatic defibrillator implantation approaches: pectoral implantation using conscious sedation reduces procedure times and cost. AB - Recent technological advances in implantable defibrillator systems (ICD) have changed implantation approaches. The aim of this study was to investigate the influence of these improvements on procedure times, implant-related charges, patient recovery, and morbidity. Ninety-six consecutive patients undergoing implantation of a nonthoracotomy ICD were studied. Implantation was performed under general anesthesia with the generator placed abdominally in 22 patients (group I) and pectorally in 40 patients (group II). Thirty-four patients underwent pectoral implantation using conscious sedation (group III). Groups were comparable with respect to clinical variables. Implantation duration and total procedure duration were shorter in group III (67 +/- 21 minutes and 117 +/- 30 minutes) when compared with group I (100 +/- 25 minutes and 157 +/- 39 minutes) and group II (86 +/- 24 minutes and 153 +/- 34 minutes, P < 0.05). Patients in group III did not require admission to the Post-Anesthesia Care Unit. In contrast, patients in groups I and II spent 92 +/- 28 minutes and 91 +/- 31 minutes in the Post-Anesthesia Care Unit. Implantation-related charges were reduced in patients having pectoral implantation using conscious sedation ($1451 +/- 217 vs. $2354 +/- 550 and $2796 +/- 384, P < 0.05). Patients in group III had a lower frequency of postoperative oral analgesic use (3.2 +/- 2.7 doses, P < 0.05) and a shortened post-operative length of stay (1.9 +/- 1.6 days, P < 0.05) when compared with groups I (5.7 +/- 4.0 doses and 3.3 +/- 1.4 days) and II (5.2 +/- 3.5 doses and 2.6 +/- 1.1 days). The overall complication rate was low (6.3%), with no differences between groups. Advances in ICD technology have simplified implantation, leading to shorter, less painful, and less expensive procedures. PMID- 9869976 TI - Effects of radiofrequency catheter ablation on patients with permanent pacemakers. AB - The objective of this study was to assess the effects of radiofrequency energy application on implanted pacemaker functions. Radiofrequency (RF) catheter ablation may cause pacemaker dysfunction due to electromagnetic interferences. The effects of RF on pacemaker behavior were studied in a series of 38 pacemakers, implanted 18 +/- 26 months prior to a RF procedure using either a right ventricular approach (AV node ablation, n = 35) or a left ventricular approach (left concealed accessory pathway ablation, n = 1; VT ablation, n = 2). The 38 patients (mean age 65 +/- 9 years) included 20 men and 18 women. Before energy applications, the 23 different pacemaker models were programmed to the VVI mode at the lowest available rate. The continuous surface ECG was recorded throughout the procedure. Thorough testing of the devices was performed before and after each RF delivery. Unusual pacemaker responses occurred in 20 of the 38 cases studied (53%). The impact of RF delivery was unpredictable, and variable dysfunctions were observed at different times for a given patient or could vary for a given model. Unusual pacemaker responses included pacemaker inhibition (n = 8), untoggled backup mode (n = 3), electromagnetic interference noise mode (n = 3), temporary RF-induced pacemaker tachycardia (n = 2), erratic behavior (n = 1), oversensing of RF onset and offset (n = 8), and transient loss of ventricular capture, (n = 1). Postablation, most devices automatically toggled back to full functionality. The three devices in the untoggled backup mode had to be reprogrammed to obtain normal operations. At the end of the procedure, pacing thresholds remained unchanged in all but one patient, in whom the increase in ventricular threshold was due to a nicked lead. In conclusion, implanted pacemakers frequently exhibit transient, unpredictable responses to RF energy application. Although all pacemaker functions were restored postablation, some devices had to be reset manually. The anomalies observed during the RF application argue for the simultaneous use of an external pacemaker in pacing dependent patients. PMID- 9869977 TI - Identification of fiber orientation in left free-wall accessory pathways: implication for radiofrequency ablation. AB - Previous reports on the anatomic discordance between atrial and ventricular insertion sites of left free-wall accessory pathways were limited and their findings were controversial. The purpose of this study was to explore the fiber orientation and related electrophysiologic characteristics of left free-wall accessory pathways. The study population comprised 96 consecutive patients with a single left free-wall accessory pathway (33 manifest and 63 concealed pathways), who underwent electrophysiologic study and radiofrequency catheter ablation using the retrograde ventricular approach. The atrial insertion site of the accessory pathway was defined from the cinefilms as the site with the earliest retrograde atrial activation bracketed on the coronary sinus catheter during tachycardia, and the ventricular insertion site was defined as the site where successful ablation of the pathway was achieved. Forty-two patients (44%) had their atrial insertion sites 5-20 mm (10 +/- 3 mm) distal to the ventricular insertion sites (proximal excursion), 30 (31%) patients had their atrial insertion sites 5-20 mm (12 +/- 3 mm) proximal to the ventricular insertion sites (distal excursion), and 24 (25%) patients had directly aligned atrial and ventricular insertion sites. Retrograde conduction properties, including 1:1 VA conduction and effective refractory period, were significantly poorer in the pathways with proximal excursion (302 +/- 67, 285 +/- 61 ms respectively) than in those with distal excursion (264 +/- 56, 250 +/- 48 ms respectively) or direct alignment (272 +/- 61, 258 +/- 73 ms respectively). Accessory pathways at the more posterior location had a significantly higher incidence of proximal excursion (P = 0.006), and those at the more anterior location had a higher incidence of distal excursion (P = 0.012). In conclusion, a wide variation in fiber orientations and related electrophysiologic characteristics was found in left free-wall accessory pathways. This may have important clinical implications for radiofrequency ablation. PMID- 9869979 TI - Clinical research in atrial fibrillation: meeting the need with a multipronged strategy. PMID- 9869978 TI - Clinical and interventional electrophysiology: a personal historical perspective. PMID- 9869980 TI - Extraction and replacement of permanent pacemaker leads through occluded vessels: use of extraction sheaths as conduits--balloon venoplasty as an adjunct. AB - Patients (pts) may present for lead extraction with symptomatic or asymptomatic subclavian vein or superior vena cava thrombosis. Replacement of permanent pacemaker leads (PPLs) in these pts may be difficult and may require accessing a new site. We examined the utility of replacing PPLs through completely occluded vessels using extraction sheaths as conduits through the total occlusion. Over six years, a total of 210 atrial and/or ventricular PPLs were extracted from 137 pts. Two pts presented with angiographically documented thrombotic occlusion of the subclavian vein. One additional pt. who had presented with a superior vena cava (SVC) syndrome, had a totally occluded innominate vein and SVC occlusion. Balloon venoplasty was used as an adjunct to dilate the SVC. In all pts, after PPLs were removed via a subclavian extraction sheath through the occluded vessel, the retained sheath was used to place a guide wire, then a peel away dilating sheath, to insert new PPLs, in each case on the side of total venous occlusion. Seven PPLs and two lead fragments were extracted, and five new PPLs replaced, ipsilateral to the venous occlusion. These data show that extraction of PPLs through thrombosed veins may be performed successfully and may not require replacing the leads through a new site. This technique spares the pt the need to access the opposite subclavian vein, and it avoids an excessive number of PPLs in the subclavian vein and SVC. The procedure illustrates an efficient means to reintroduce new PPLs with the potential to reduce associated morbidity, since repeat puncture of the subclavian vein is not required. Safety of the procedure as a whole must be considered with regard to the known risks of lead extraction, some complications of which may be substantial using current techniques. PMID- 9869981 TI - Programmed stimulation of the ventricles--back to the future. PMID- 9869982 TI - Atrioventricular nodal reentrant tachycardia with atrioventricular block. AB - Atrioventricular block (AVB) during atrioventricular nodal reentrant tachycardia (AVNRT) has been well documented, although it is not a common phenomenon. The mechanism for the initiation and resolution of AVB during AVNRT have been postulated. However, the site of AVB and its implication on the reentrant circuit in AVNRT is not clear. We illustrate two examples of AVNRT with AVB and offer further clarification on the site and mechanism of AVB. PMID- 9869983 TI - Pharmacologic enhancement of atrial electrical defibrillation efficacy: role of ibutilide. AB - The study was designed to identify the influence of ibutilide on activation during atrial fibrillation (AF) and determine its role in enhancing the efficacy of electrical conversion of AF. Vagally-mediated acute AF was induced in 12 anesthetized dogs. In 5 dogs, a cross-shaped epicardial patch containing 16 recording electrodes was placed on the right atrium. In 7 dogs, defibrillation patch electrodes overlying both atria were used to deliver biphasic shocks (50% tilt; 90 microF). Measurements were made at baseline and following ibutilide (0.075 mg/kg bolus followed by 0.075 mg/kg infusion over 10 minutes). Right atrial multisite electrogram recordings revealed significant prolongation in AF cycle length at all sites following ibutilide. In all dogs, AF cycle length increased with ibutilide from 90 +/- 23 to 130 +/- 49 ms (p < 0.005). Whereas, atrial defibrillation threshold decreased from 0.83 +/- 0.5 to 0.53 +/- 0.29 J (p = 0.020). In conclusion, ibutilide prolongs cycle length of canine vagally mediated acute AF. Furthermore, ibutilide facilitates electrical conversion of AF by lowering energy requirement. Thus, controlled drug infusion in conjunction with electrical defibrillation may be useful for managing AF. PMID- 9869984 TI - A randomized comparison of fixed power and temperature monitoring during slow pathway ablation in patients with atrioventricular nodal reentrant tachycardia. AB - Temperature monitoring may be helpful for ablation of accessory pathways, however its role in ablation of atrioventricular nodal reentrant tachycardia (AVNRT) using the slow pathway approach is unclear. Therefore, the purpose of this study was to prospectively compare slow pathway ablation for AVNRT using fixed power or temperature monitoring. The study included 120 patients undergoing ablation for AVNRT. Patients were randomly assigned to receive either fixed power at 32 watts, or to temperature monitoring with a target temperature of 60 degrees C. The primary success rate was 72% in the fixed power group and 95% in the temperature monitoring group (p = 0.001). The ablation procedure duration (35 +/- 29 min vs 35 +/- 30 min; p = 0.9), fluoroscopic time (32 +/- 17 vs 35 +/- 19 min; p = 0.4), mean number of applications (10.2 +/- 8.1 vs 8.4 +/- 7.9; p = 0.2), and coagulum formation per application (0.2% vs 0.5%; p = 0.6) were statistically similar in the fixed power and temperature monitoring groups, respectively. The mean temperature (47.3 +/- 4.8 degrees C vs 48.6 +/- 3.8 degrees C; p < 0.01), and the temperature associated with junctional ectopy (48.2 +/- 3.8 degrees C vs 49.3 +/- 3.6 degrees C, p < 0.01) were less for the fixed power than the temperature monitoring group. In the temperature monitoring group, only 31% of applications achieved an electrode temperature of 60 degrees C. During follow up of 6.6 +/- 3.6 months there were two recurrences in the fixed power group and one in the temperature monitoring group (p = 1.0). In summary, power titration directed by temperature monitoring was associated with an improved primary procedural success rate. Applications of energy were associated with a temperature of approximately 50 degrees C with both techniques, suggesting that there is a low efficiency of heating in the posterior septum. PMID- 9869985 TI - Atrioventricular node reentrant tachycardia in patients with a prolonged AH interval during sinus rhythm: clinical features, electrophysiologic characteristics and results of radiofrequency ablation. AB - Among a consecutive series of 600 patients who underwent radiofrequency catheter ablation for AV node reentrant tachycardia, 14 patients (age 29-76 years) had a prolonged AH interval during sinus rhythm (172 +/- 18 ms, range 140 to 200). Seven of them had unsuccessful ablation during the previous ablation sessions. Eight patients with anterograde dual AV node pathway physiology received anterograde slow pathway ablation, and the other 6 patients without dual-pathway physiology received retrograde fast pathway ablation. All patients had successful elimination of AV nodal reentrant tachycardia after a mean of 4 +/- 4 radiofrequency applications, power level 36 +/- 6 watts and a pulse duration of 42 +/- 4 seconds. The postablation AH interval remained unchanged. During a follow-up period of 25 +/- 13 months, one patient who received slow pathway ablation developed 2:1 AV block with syncope. As compared with the other 586 patients without a prolonged AH interval, these 14 patients had significantly poorer anterograde AV nodal function and lower incidence of anterograde dual AV node physiology (P < 0.01). We concluded that slow pathway ablation in patients with dual pathway physiology, and retrograde fast pathway ablation in patients without dual pathway physiology were effective and safe in patients with a prolonged AH interval. However, delayed onset of symptomatic AV block is possible and careful follow-up is necessary. PMID- 9869986 TI - Recurrence of accessory pathway conduction after successful radiofrequency ablation: histological findings. AB - The histological findings from a patient with Wolff-Parkinson-White syndrome (right superior paraseptal accessory pathway) who underwent successful radiofrequency ablation and had recurrence of tachycardia one month later in the absence of overt pre-excitation are reported. Histology revealed three small, oval to circular shaped, whitish, smooth areas on the right endocardial surface, the one being situated at the atrial free wall, and the other two being at the ventricular aspect. A very small hole was present in the interventricular component of the membranus septum. The accessory pathway band passed to either side of the small hole albeit disrupted by fibrous tissue in places. These findings indicate that multiple applications may cause penetration of the myocardium, and to achieve success, complete disruption of the pathway at some point along its course is required. Recurrence of retrograde accessory pathway conduction can be explained with the impedance mismatch hypothesis. PMID- 9869988 TI - Torsade-de-pointes. PMID- 9869989 TI - The use of ICD's for the treatment of patients with Arrhythmogenic Right Ventricular Dysplasia (ARVD) PMID- 9869987 TI - Advances in cardiology: the complementary roles of concept and technology. PMID- 9869990 TI - Evaluating antiarrhythmic drugs in implantable device. PMID- 9869991 TI - Familial hypertrophic cardiomyopathy mice display gender differences in electrophysiological abnormalities. AB - Genetically-manipulated mice harboring an alpha-myosin heavy chain Arg403Gln missense mutation (alpha-MHC403/+) display a phenotype characteristic of familial hypertrophic cardiomyopathy (FHC). Male and female (30 +/- 8 week old) heterozygous alpha-MHC403/+ mice and litter-mate controls were evaluated using a surface electrocardiogram (ECG) and an in vivo cardiac electrophysiology study (EPS). Wild type animals had normal intracardiac electrophysiology, with no significant differences between male and female control mice during EPS. The female wild-type mice did have slower heart rates and longer ECG intervals than their male wild-type counterparts. The female alpha-MHC403/+ mice had similar ECG's, cardiac conduction times, and refractory periods compared with female wild type mice. In contrast, male FHC mice had distinctive ECG and electrophysiologic abnormalities including right axis deviation, prolonged ventricular repolarization and prolonged sinus node recovery times. During programmed ventricular stimulation, 62% of male alpha-MHC403/+ mice and 28% of female alpha MHC403/+ mice had inducible ventricular tachycardia. These studies identify gender-specific electrophysiologic abnormalities in alpha-MHC403/+ FHC mice, concordant with the histological and hemodynamic derangements previously reported. PMID- 9869992 TI - Atrial electrical remodeling by rapid pacing in the isolated rabbit heart: effects of Ca++ and K+ channel blockade. AB - INTRODUCTION: Electrical remodeling describes atrial electrophysiologic changes that occur following atrial fibrillation. The mechanism(s) responsible for this phenomenon is not well understood. The purpose of this study was to examine the effects of rapid atrial pacing on atrial action potential duration, conduction time and refractoriness in the isolated rabbit heart. The effects of Ca++ and K+ blockade in this model were also studied. METHODS AND RESULTS: Monophasic action potential recordings were made from 12 epicardial atrial sites in 50 isolated perfused rabbit heart preparations. These recordings were analyzed for activation time (AT), 90% action potential duration (APD) and conduction times (CT) measured at a 250 msec cycle length. Atrial effective refractory periods (ERP) were determined at a 200 msec cycle length. All measurements were made at baseline and repeated after 2 hours of biatrial pacing at 250 msec (control group, n = 10) or 2 hours of rapid biatrial pacing (approximately 80 msec) in 4 groups: rapid pacing alone (rapid pacing group); rapid pacing in the presence of 0.1 mM verapamil (verapamil group) for L-type Ca++ channel blockade; rapid pacing with 1 mM 4-aminopyridine (4-AP group) for K+ channel blockade; and rapid pacing with 50 microM nickel chloride (Ni++ group) for T-type Ca++ channel blockade (n = 10 each group). All baseline and post pacing measurements were taken in the presence of Ca++ or K+ blockers for the respective groups. After rapid atrial pacing alone the average APD shortened by 8.2 +/- 10.4 msec compared to 3.6 +/- 12.5 msec shortening for control group (p = 0.002). The shortening of APD was uniform at all recording sites. For the rapid pacing group, CT was unchanged for right to left atrial conduction but shortened significantly for left to right atrial conduction (26.8 +/- 1.9 msec at baseline to 22.3 +/- 4.1 msec post pacing, p = 0.005). Conduction times were unchanged in the control group. The dispersion of repolarization was unchanged by rapid pacing alone. The decrease in APD from baseline to post rapid pacing was similar to the control group for those hearts treated with verapamil and 4-AP (1.5 +/- 12.3 and 4.7 +/- 10.4 msec, respectively, both p > or = 0.18 vs control group). The decrease in APD was significantly greater for the Ni++ group (11.8 +/- 14.3 msec) than for either the control group or rapid pacing group (both p < or = 0.023). The dispersion of repolarization was increased only in the 4-AP group post rapid pacing (41.7 +/- 6.2 msec at baseline to 53.5 +/- 9.6 msec post pacing, p = 0.01). ERPs were unchanged in any of the 5 groups except for a decrease in left atrial ERP in the Ni++ group after rapid pacing (98 +/- 14 msec at baseline to 88 +/- 8 msec post rapid pacing, p = 0.005). CONCLUSIONS: In the isolated rabbit heart model: 1) atrial APD is shortened after rapid pacing; 2) the shortening of APD is attenuated by verapamil and 4-AP but exaggerated by Ni++; 3) atrial conduction times are shortened in a direction specific manner after rapid pacing; and 4) shortening of ERP in this model is measured only in the presence of Ni++. These findings suggest that both L-type Ca++ and 4-AP sensitive channels may participate in atrial electrical remodeling. PMID- 9869993 TI - Review article: heart rate and blood pressure control in vasovagal syncope. AB - Vasovagal syncope is characterized by transient failure of usually reliable physiologic mechanisms responsible for maintaining both systemic arterial pressure and cerebral blood flow. Two circulatory phenomena are almost universally present: systemic arterial vasodilation and bradycardia. A third phenomenon, cerebrovascular constriction, has also been described but its contribution to the faint is less well established. The neural reflex pathways responsible for triggering the circulatory changes in the vasovagal faint are incompletely understood, but have recently been the subject of renewed interest. In part, this interest probably stems from the frequency with which vasovagal symptoms are now recognized to be the cause of fainting spells. Additionally, however, there is an increasingly recognized need to develop treatment strategies for those affected patients in whom recurrent vasovagal symptoms are particularly troublesome. It is the goal of this discussion to focus on those aspects of circulatory control, and in particular on potential interactions among certain neural and humoral systems, which may contribute to the inappropriate physiologic responses associated with the vasovagal faint. PMID- 9869994 TI - Unipolar sensing in contemporary pacemakers: using myopotential testing to define optimal sensitivity settings. AB - Bipolar lead use has increased due to oversensing concerns with older unipolar systems. Data on contemporary unipolar devices with improved hardware design and greater programming flexibility is lacking. Using a randomized crossover design, unipolar and bipolar sensing characteristics of 22 atrial and 16 ventricular leads were compared in 34 patients who had pulse generators of programmable polarity. Unipolar and bipolar intracardiac electrogram amplitudes, pacing and sensing thresholds at rest were similar. Provocative maneuvers were used to assess for myopotential inhibition. At atrial sensitivities of 0.625-1.50 mV, myopotential inhibition occurred in 11 (50%) atrial leads in the unipolar mode compared to 1 (5%) in the bipolar mode (p < 0.001). At sensitivities of > 1.50 mV myopotential inhibition occurred in only 1 ventricular (unipolar) lead. An optimal sensitivity setting for each polarity was derived using clinic test results and assessed by ambulatory ECG (AECG). At these optimal settings, oversensing occurred in 1 (6%) atrial and 1 (8%) ventricular unipolar lead during AECG monitoring, whereas oversensing was not seen in any leads programmed to the bipolar mode. Undersensing occurred in 5 (29%) atrial unipolar versus 1 (6%) bipolar lead (p = 0.08). Undersensing was not observed in any of the ventricular leads. Myopotential inhibition may be frequently provoked by provocative maneuvers at higher sensitivity settings in atrial unipolar leads. The frequency of oversensing can be significantly reduced by defining an optimal sensitivity setting using simple isometric maneuvers. Given present day concerns over bipolar lead longevity, increased utilization of unipolar ventricular leads should be considered. PMID- 9869995 TI - Randomized comparison of a 90 uF capacitor three-electrode defibrillation system with a 125 uF two-electrode defibrillation system. AB - INTRODUCTION: A variety of factors, including the number of defibrillation electrodes and shocking capacitance, may influence the defibrillation efficacy of an implantable defibrillator system. Therefore, the purpose of this study was to compare the defibrillation energy requirement using a 125 uF two-electrode defibrillation system and a 90 uF three-electrode defibrillation system. METHODS AND RESULTS: The defibrillation energy requirements measured with both systems were compared in 26 consecutive patients. The two-electrode system used a single transvenous lead with two defibrillation coils in conjunction with a biphasic waveform from a 125 uF capacitor. The three-electrode system used the same transvenous lead, utilized a pectoral implantable defibrillator generator shell as a third electrode, and delivered the identical biphasic waveform from a 90 uF capacitor. The two-electrode system was associated with a higher defibrillation energy requirement (10.8 +/- 5.5 J) than was the three-electrode system (8.9 +/- 6.7 J, p < 0.05), however, the leading edge voltage was not significantly different between systems (361 +/- 103 V vs. 397 +/- 123 V, P = 0.07). The two electrode system also had a higher shocking resistance (49.0 +/- 9.0 ohms vs. 41.4 +/- 7.3 ohms, p < 0.001) and a lower peak current (7.7 +/- 2.6 A vs. 10.1 +/ 3.7 A, p < 0.001) than the three-electrode system. CONCLUSIONS: A three electrode defibrillation system that utilizes a dual coil transvenous lead and a subcutaneous pectoral electrode with lower capacitance is associated with a lower defibrillation energy requirement than is a dual coil defibrillation system with higher capacitance. This finding suggests that the utilization of a pectoral generator as a defibrillation electrode in conjunction with smaller capacitors is a more effective defibrillation system and may allow for additional miniaturization of implantable defibrillators. PMID- 9869996 TI - Submuscular versus subcutaneous pectoral implantation of cardioverter defibrillators: effect on high voltage pathway impedance and defibrillation efficacy. AB - Implantable cardioverter-defibrillator (ICD) pulse generators are now routinely positioned in a pectoral location, either submuscularly (under the pectoralis muscles) or subcutaneously (over the pectoralis muscles). Furthermore, in current ICDs, the generator shield usually participates in the defibrillation energy pathway ("hot can"). Consequently, the precise generator location could affect defibrillation system efficacy. To assess this issue, we compared high voltage pathway impedance and defibrillation threshold (DFT) in 20 patients undergoing submuscular and 46 patients undergoing subcutaneous pectoral implantation of an Angeion Sentinel ICD and an AngeFlex dual-coil defibrillation lead. Measurements were performed at time of ICD implant, pre-hospital discharge, and 1, 3 and/or 6 months later. Following induction of ventricular fibrillation, 569 biphasic waveform shocks were delivered between the generator shield and either the distal defibrillation coil (RV/can configuration) or both proximal and distal coils (RV/SVC/can configuration). Impedance differences between submuscular and subcutaneous implants were approximately 3-4 Ohms (p value of 0.132 to < 0.001 depending on time of follow-up and lead configuration). A significant increase in impedance over time was noted independent of implant location and lead configuration. The DFT at implant or pre-discharge was assessed in 27 individuals, and was 9.9 +/- 3.8 J in 8 patients in the submuscular group, and 7.4 +/- 3.3 J in 19 patients in the subcutaneous group (p = 0.057). In conclusion, anatomic location of a "hot can" ICD generator (submuscular versus subcutaneous) influences impedance to defibrillation current, but the impact is of small magnitude and does not appear to result in clinically important differences in DFT. PMID- 9869997 TI - Case report: inappropriate discharge of an implantable cardioverter-defibrillator caused by the combined count criterion. AB - We describe a rare case of an inappropriate defibrillator discharge during a sinus tachycardia with single fast sensed beats fulfilling the ventricular fibrillation rate criterion, activating the "combined count criterion," although the prevalent sinus tachycardia was inside a ventricular tachycardia rate zone, programmed to monitor only mode. The "combined count criterion" is a safety feature that ensures defibrillation therapy in case of a tachyarrhythmia inside the ventricular tachycardia detection zone, that is temporarily crossing the ventricular fibrillation detection rate. This case reports shows that there is a low but unpredictable risk of device therapy even in a zone programmed to monitor only mode. This phenomenon should be considered when a monitor only zone is installed for revelation of slower tachyarrhythmias. PMID- 9869998 TI - Inferior vena cava-tricuspid annulus isthmus is a critical site of unidirectional block during the induction of common atrial flutter. AB - BACKGROUND: Until recently no clinical studies had reported precise right atrium (RA) mapping when performing induction of atrial flutter (AFl). We studied the mode of tachycardia initiation in 16 patients (pts) referred for radiofrequency (RF) AFl ablation. AFl induction was performed at the beginning of the procedure (n = 10), or after previous AFl termination during RF delivery (n = 6). Detailed analysis of AFl initiation was provided by duodecapolar (Halo) and multipolar catheters positioned in the peritricuspidian region at the lateral right atrial wall (LRA), the inferior vena cavatricuspid annulus (IVC-TA) isthmus and the interatrial septum. Induction was obtained during incremental pacing (IAP) (15 pts) or programmed stimulation (1 pt) from the proximal coronary sinus (PCS). RESULTS: Atrial flutter with counterclockwise (CCW) RA rotation was induced in all pts by PCS pacing. During PCS IAP, at long pacing cycle lengths, impulse propagated in a clockwise (CW) direction through the IVC-TA isthmus and then upward at low (L) LRA. This led to a collision at the mid LRA with another wave front propagating in a CCW direction at the septum. IAP from PCS induced a progressive delay of propagation at the IVC-TA isthmus resulting in a prolongation of the PCS-Mid Isthmus interval from 85 +/- 29 to 151 +/- 42 msec. At same pacing cycle lengths (CL), the PCS-HLRA interval was comparatively less prolonged, from 75 +/- 12 to 105 +/- 18 msec, p = 0.0007. This preferential slowing of conduction between PCS and mid isthmus, during IAP from PCS, was associated with a displacement of the zone of collision to the Low LRA. Finally a CW functional block occurred at the IVC-TA isthmus and CCW AFl was induced through a period of transient concealed entrainment. The paced CL required to initiate flutter ranged from 290 to 180 msec and the mean CL of induced atrial flutter was 254 +/- 27 msec. CONCLUSIONS: The IVC-TA isthmus has decremental properties and exhibits wenckebach phenomenon during incremental PCS pacing. Initiation of a counterclockwise flutter by PCS pacing is associated with appearance of a functional unidirectional block at the IVC-TA isthmus. PMID- 9869999 TI - Ventricular performance and quality of life in patients who underwent radiofrequency AV junction ablation and permanent pacemaker implantation due to medically refractory atrial tachyarrhythmias. AB - In patients with drug refractory atrial tachyarrhythmias and previous failed attempts of ablation of the arrhythmia substrate, radiofrequency (RF) modulation or ablation of the atrioventricular (AV) junction is an alternative procedure. AIM: Of this study was to assess the efficacy and long term results of RF AV junction ablation in conjunction with permanent pacemaker implantation, in the management of patients with drug resistant atrial tachyarrhythmias. METHODS: Between 4/92 and 1/97, 46 patients (30 male, 16 female, 67 +/- 12 years) underwent RF AV junction ablation because of paroxysmal atrial fibrillation (24 patients), chronic atrial fibrillation (13 patients), atrial flutter (5 patients) and atrial tachycardia (4 patients). The underlying heart disease was dilated cardiomyopathy (16), ischemic heart disease (9), hypertensive heart disease (6), hypertrophic cardiomyopathy (3), atrial septal defect (2) and non structural heart disease (10). The duration of symptoms was 6.4 +/- 3.5 years at a maximal heart rate 169 +/- 24 bpm. The hospital admissions in the last 12 months were 8.2 +/- 3 per patient. The failed antiarrhythmic drugs were 3.5 +/- 2.1. The functional NYHA class was 2.7 +/- 0.6. Patients with atrial flutter and atrial tachycardia had previous failed attempts of RF ablation of the arrhythmia substrate. Thirty patients had a compromised left ventricular systolic function with LVEF below 50% (mean 34 +/- 9%). AV junction ablation was achieved in all patients after 4 +/- 2.5 RF applications. Post ablation, the selected pacing mode was DDD-R for the 33 patients with paroxysmal atrial tachyarrhythmias and VVI-R for the 13 pts with chronic atrial fibrillation. The dual chamber pacemakers implanted had the option of automatic mode switch. RESULTS: During the follow-up period of 28 +/- 13 months (6-47), AV conduction recovered in 1 patient. Antiarrhythmic treatment was necessary in only 7 patients. Post ablation the new functional NYHA class was 1.4 +/- 0.8 (p < 0.001). Post ablation hospital admissions, including ordinary pacemaker follow-up visits, were 4 +/- 1 per patient per year (p < 0.001). Six months after the procedure the LVEF of the study population was increased from 42 +/- 16% to 50 +/- 14% (p = NS). In the 30 patients with heart failure the LVEF was significantly increased to 46 +/- 8% (p < 0.05). Symptomatic relief or significant improvement was observed in all patients as showed by the answers given in a customized questionnaire before and after the procedure. CONCLUSIONS: In patients with drug refractory atrial tachyarrhythmias, RF AV junction ablation and permanent pacemaker implantation is an alternative therapy with excellent long term results in terms of arrhythmia control, ventricular performance and quality of life. PMID- 9870001 TI - Case report: radiofrequency catheter ablation of atrioventricular nodal reentrant tachycardia in a patient with heart transplantation. AB - Ten years after orthotopic cardiac transplantation, a 56-year-old man developed recurrent presyncope and syncope. A 24-hour ambulatory electrocardiographic recording did not document significant arrhythmic events. A head-up tilt table test was negative. An electrophysiologic study revealed dual atrioventricular (AV) nodal physiology and inducible typical atrioventricular nodal reentrant tachycardia (AVNRT). The patient became hypotensive and presyncopal during AVNRT. Radiofrequency (RF) catheter ablation successfully eliminated AVNRT without complications. The patient remained free of symptoms at nine months follow-up. PMID- 9870000 TI - Single radiofrequency application to cure atrioventricular nodal reentry: arguments for the slow pathway origin of the high-low frequency slow potentials. AB - BACKGROUND: High-low frequency slow potentials are thought to be related to the slow AV pathway conduction. Their use was proposed to guide radiofrequency (RF) ablation of atrioventricular nodal reentrant tachycardia (AVNRT). The present study was designed to determine the prospective value of these high-low frequency slow potentials to guide AVNRT ablation using a single RF application. Single RF application could indeed reduce the size of the lesion created in the viciny of the specialized AV conduction system and shorten the radiation exposure and the overall duration of the procedure. RESULTS: Forty-one patients (14 men, 27 women, 45 +/- 16 years old) with AVNRT underwent slow pathway RF ablation guided by high low frequency slow potentials. High-low frequency slow potentials were found in all patients along the tricuspid annulus and above the coronary sinus. Ablation was always performed in the posterior part of Koch's triangle. The mean A/V amplitude ratio of the successful site was 0.43 +/- 0.59. In 32 patients (78%) AVNRT was no longer inducible after a single RF application. Procedure and radiation times were 35 +/- 31 and 13 +/- 12 min respectively. Five patients required 2, 3 patients 3, and 1 patient 6 RF applications. The mean number of RF applications was 1.4 +/- 0.9 (median = 1). In the 32 patients who required only one RF application, 24 (75%) had an obvious dual AV nodal pathways with a jump before ablation, which completely disappeared in 18 of them (75%) after ablation. In the 6 remaining patients, who still had a jump after 1 RF application, there was no significant change in either conduction times or refractory periods concerning both the anterograde and retrograde AV conduction. No patient had PR interval purlongation. After a mean follow up of 11 +/- 5 months, recurrence was observed in a single patient who received 2 discontinued RF applications. CONCLUSION: Catheter-mediated ablation of AVNRT using high-low frequency slow potentials to localize the slow AV pathway is feasible and safe. Using this technique, a single RF application was successfull in 78% of patients, and slow pathway characteristics were completely eliminated in 75% of patients. The radiation time and the procedure duration were short. This suggest that, in patients with AVNRT, the choice of an appropriate RF target can reduce procedural duration. PMID- 9870002 TI - Neonatal radiofrequency catheter ablation of junctional tachycardias. AB - Junctional tachycardias comprise several arrhythmia types with differing mechanisms, principally involving the region of the atrioventricular (A-V) junction. Neonatal radiofrequency catheter ablation has typically been reserved for life-threatening, drug-refractory cases due to the unique concerns regarding patient size and development. We performed radiofrequency catheter ablation on two neonates with incessant, rapid junctional tachycardias and hemodynamic compromise after failing conventional medical therapy. This report describes 2 neonates who underwent emergent radiofrequency catheter ablation, and compares these two patients to a larger pediatric catheter ablation patient cohort. Both neonates had an acutely successful outcome and were able to be discharged within a week of the ablation procedure. Fluoroscopy time and total procedure time were shorter in these two patients than in the course of the average pediatric catheter ablation. Though long-term developmental consequences of neonatal catheter ablation are yet unknown, in unique extreme situations, radiofrequency catheter ablation can be performed in neonates, as in older children and adults, without excessive acute morbidity. PMID- 9870003 TI - Teaching interventional electrophysiology: is revision of core curricula in order? PMID- 9870005 TI - Be still my beating heart--the Ablate and Pace Study. PMID- 9870004 TI - The Ablate and Pace Trial: a prospective study of catheter ablation of the AV conduction system and permanent pacemaker implantation for treatment of atrial fibrillation. APT Investigators. AB - BACKGROUND: The Ablate and Pace Trial (APT) prospectively assessed the effects of catheter ablation of the AV conduction system and permanent pacemaker implantation on health-related quality of life, survival, exercise capacity, and ventricular function in 156 patients with symptomatic atrial fibrillation. METHODS: All patients referred for catheter ablation and permanent pacemaker implantation because of medically-refractory atrial fibrillation at 16 centers were screened for enrollment in a prospective registry. Baseline assessment prior to ablation included measurement of quality of life, including the Health Status Questionnaire, the Quality of Life Index and the Symptom Checklist: Frequency and Severity. Exercise capacity was assessed with metabolic treadmill exercise testing and ventricular function was quantitated with echocardiography. The quality of life instruments, exercise capacity, and echocardiography were repeated at 3 and 12 months after catheter ablation. RESULTS: The APT population included 90 men and 66 women (66.1 +/- 11.5 years of age) with either chronic (n = 70), recurrent (n = 31), or paroxysmal atrial fibrillation (n = 55). Structural heart disease was present in 78.2% of patients. Successful ablation of AV conduction was achieved in 155 of 156 patients (99.4%). Survival at 1 year was 85.3%, with 5 of 23 deaths being sudden cardiac deaths. Survival over the first year of follow-up was significantly lower for patients with a baseline left ventricular ejection fraction (LVEF) < 0.45 (0.73) than for patients with a LVEF > or = 0.45 (0.88, p = 0.03). The NYHA functional class improved from 2.1 at baseline to 1.8 at 3 months and 1.9 at 12 months of followup (p = 0.0001). Significant improvement in quality of life scores were noted for all 8 subscales of the Health Status Questionnaire, for the overall rating of the Quality of Life Index, the Health and Function subscales; Arrhythmia-related symptoms were markedly reduced as measured by the Symptom Checklist: Frequency and Severity scale. The mean LVEF improved from 0.50 +/- 0.20 at baseline to 0.54 +/- 0.20 at 3 months (p = 0.03). The LVEF 12 months after ablation was 0.52 +/- 0.20, not statistically different from baseline. Individuals with reduced systolic function at baseline had the greatest improvement, from LVEF 0.31 +/- 0.20 at baseline to 0.41 +/- 0.20 at 3 months and 0.41 +/- 0.30 at 12 months (p = 0.0001). There were no significant changes in treadmill exercise duration (10.0 +/- 4.3 min at baseline and 11.6 +/- 3.6 min at 12 months) or VO2max (1467 +/- 681 ml O2 min baseline and 1629 +/- 739 ml O2 min at 12 months). CONCLUSIONS: Catheter ablation of the AV conduction system and permanent pacemaker implantation were associated with improved quality of life and left ventricular function in this population of highly symptomatic patients with atrial fibrillation refractory to medical therapy. PMID- 9870007 TI - Phase 1B ventricular arrhythmia in the dog: localized reentry within the mid myocardium. AB - Intramural and epicardial composite electrograms, signal-averaged orthogonal bipolar electrograms across the ischemic zone, and closely-spaced bipolar electrograms from subendocardium, mid-myocardium, and subepicardium were utilized to determine if phase 1B reentry resulted from localized reentry within ischemic mid-myocardium. During the first 10 minutes following coronary ligation, activation delays were largest in ischemic subepicardium, with continuous electrical activity in ischemic epicardium linking a ventricular extrasystole to the preceding beat. During the 15-30 minute period, activation delay observed in ischemic mid-myocardium exceeded activation delay on the epicardial surface. Ventricular extrasystoles were associated with mid-myocardial delays > 130 msec. With short-coupled extrasystoles (< 300 msec), electrical activity in ischemic mid-myocardium linked an extrasystole with the preceding beat. Although single extrasystoles with coupling intervals > 300 msec were also associated with mid myocardial delays > 130 msec, most extrasystoles (68%) demonstrated an isoelectric gap > 20 msec. The data demonstrate an association between delayed activation within ischemic mid-myocardium and phase 1B arrhythmia resulting from (1) localized reentry in ischemic mid-myocardium and (2) a delay-dependent "non reentrant" mechanism. PMID- 9870006 TI - Design and implementation of the Dual Site Atrial Pacing to Prevent Atrial Fibrillation (DAPPAF) clinical trial. DAPPAF Phase 1 Investigators. AB - The Dual Site Atrial Pacing to Prevent Atrial Fibrillation (DAPPAF) study compares dual site, single site and support pacing modalities in the prevention of atrial fibrillation (AF) in patients with a history of paroxysmal AF (PAF) and a bradyarrhythmic indication for pacing. The trial is a randomized crossover comparison of dual site atrial pacing, single site atrial pacing, and a support pacing control period (DDI at 50 ppm or VDI) done in six month intervals. Patient inclusion requires at least 2 documented AF episodes in the three months prior to enrollment. The patients can be on concurrent antiarrhythmic drug regimens but this regimen must remain constant throughout the protocol. Patients with AV nodal ablation are excluded from this study. The primary endpoints of the study compare the time to first recurrence of clinically significant symptomatic AF with ECG verification, and quality of life among the three treatment modes. Secondary endpoints include time to first recurrence of all AF episodes as monitored by the pacemaker, the measurement of echocardiographic parameters, and symptoms logged by the patients. This trial was designed after pilot studies showed dual site pacing to be safe, feasible and preliminary results suggested increased maintenance of sinus rhythm with atrial pacing. PMID- 9870008 TI - Coronary blood flow changes during atrioventricular sequential pacing with different atrioventricular delays in normal individuals. AB - This study examined the effects of different atrioventricular (AV) intervals, during AV sequential pacing, on hemodynamics and coronary blood flow in individuals with normal hearts. Left anterior descending artery blood flow velocity was measured, using intracoronary Doppler, in 17 normal individuals. Five pacing tests were applied in random order for 5 min, at 15 beats/min above the sinus rate. Four tests using AV sequential pacing with AV intervals of 175, 150, 100, and 50 ms, and one using atrial pacing were applied. Mean flow velocity was 21 +/- 9 cm/s, 20 +/- 9 cm/s, 17 +/- 7 cm/s, 17 +/- 7 cm/s, and 22 +/- 10 cm/s, respectively (F = 8.87, p = .00001). The hemodynamic effects of these 5 pacing tests were assessed in 8 different normal subjects. Isovolumic relaxation time constant and left ventricular systolic pressure decreased, whereas right atrial pressure increased during AV sequential pacing with short AV intervals. Thus, during short-term AV sequential pacing at rest, coronary blood flow in a normal left anterior descending artery decreases with short AV intervals. PMID- 9870009 TI - Acute evaluation of a preformed single-pass VDD/DDD pacing lead. AB - The clinical utility of single lead, atrial synchronous, ventricular pacing (VDD), for patients with normal sinus function and heart block is well established. Atrial stimulation, unavailable in VDD systems presents a significant disadvantage. DDD pacing systems however, require the introduction and positioning of two separate leads. The acute human study discussed evaluated a modified version of a commercially available VDD lead with a preshaped lobe, capable of both sensing and pacing the right atrium. "P" waves and atrial stimulation thresholds were determined in five patients. The mean P-wave was 2.5 +/- 1 millivolts. Atrial stimulation in the unipolar configuration was 1.6 +/- 0.5 volts and 1.7 +/- 0.9 volts in the bipolar configuration. These acute stimulation thresholds and sensing amplitudes were comparable to conventional DDD pacing systems. Therefore, single-lead DDD pacing may be feasible and warrants further study. PMID- 9870010 TI - Complications of dual chamber pacemaker implantation in the elderly. Pacemaker Selection in the Elderly (PASE) Investigators. AB - Pacemakers are frequently implanted, yet accurate prospective data on implant complications are limited. Elderly patients may be at increased risk of implant complications and are increasingly being referred for pacemaker implantation. The purpose of the present analysis was to define the incidence and possible predictors of serious complications of dual chamber permanent pacemaker implantation in the elderly. Therefore, we sought to prospectively identify the incidence and predictors of pacemaker implant complications in a large multicenter trial involving patients receiving a dual chamber pacemaker. The Pacemaker Selection in the Elderly (PASE) study was a prospective trial designed to evaluate quality of life in dual chamber pacemaker recipients age 65 years or older randomized to DDDR versus VVIR programming. In addition to being age 65 years or older, patients enrolled in this study were in normal sinus rhythm, and had standard indications for permanent pacemaker implantation. All patients received dual chamber pacemakers and were randomized to DDDR versus VVIR pacing. Pacemaker implant complications were collected on standardized forms which were completed at pacemaker implantation and during follow-up appointments. In this study of 407 patients, there were 26 complications occurring in 25 patients (6.1%). The most frequent complication was lead dislodgment which occurred in 9 patients. This was followed by pneumothorax (8 patients) and cardiac perforations (4 patients). In 18 patients (4.4%) repeat surgical procedures (including chest tubes) were required. Complications were noted prior to discharge in only 18 patients. There were no significant predictors of overall complications. Pneumothorax was more frequent in patients > or = 75 years old, and was observed only in patients with subclavian venous access. In conclusion, complications from pacemaker implantation in the elderly are seen in 6.1% of patients and 4.4% of patients require a repeat surgical procedure. Other than advanced age and lower weight predicting for pneumothorax, there are no significant clinical predictors of complications. PMID- 9870011 TI - Electrophysiologic characteristics of a dilated atrium in patients with paroxysmal atrial fibrillation and atrial flutter. AB - This study investigated the difference of atrial electrophysiologic characteristics between a normal and dilated atrium and compared them among patients with paroxysmal atrial fibrillation and flutter. Twenty-seven patients with paroxysmal atrial fibrillation and 28 patients with paroxysmal atrial flutter were divided into four subgroups, according to the presence of a normal atrium or bilateral atrial enlargement. Thirty patients without atrial arrhythmia (20 patients with normal atrium and 10 patients with bilateral atrial enlargement) were included in control group. The atrial refractoriness in patients with a dilated atrium was longer than those with normal atrial size. In patients with paroxysmal atrial fibrillation and patients of control group, the P wave duration and interatrial conduction velocity with or without atrial enlargement were similar. However, in patients with paroxysmal atrial flutter, P APCS (86 +/- 10 ms vs. 73 +/- 9 ms, p < 0.05) and P-ADCS (109 +/- 9 ms vs. 95 +/- 9 ms, p < 0.05) in patients with a dilated atrium were longer than in patients with a normal atrium. The patients with paroxysmal atrial fibrillation or atrial flutter all demonstrated longer P-wave duration and interatrial conduction time than control group. Among the groups with a normal atrium or a dilated atrium, atrial refractoriness in patients with paroxysmal atrial flutter was shorter than that in control group. Moreover, in the patients with a normal atrium, the potential minimal wavelength in control group (6.6 +/- 1.7) was longer than that of paroxysmal atrial fibrillation (5.3 +/- 1.1), or atrial flutter (5.0 +/- 1.2). These findings suggest that atrial electrophysiologic characteristics of a dilated atrium were different from those of normal atrium, and these changes were different between paroxysmal atrial fibrillation and flutter. Multiple factors are considered to be related to the genesis of atrial tachyarrhythmias. PMID- 9870012 TI - Idiopathic left ventricular aneurysm: an unusual substrate of ventricular fibrillation and ventricular tachycardia. AB - Idiopathic left ventricular aneurysm (LVA) is a very rare clinical condition. This article describes a patient with idiopathic LVA associated with episodes of ventricular tachycardia and ventricular fibrillation. Clinical and instrumental examinations did not reveal the pathogenesis of the aneurysm. The malignant clinical course suggests that an aggressive antiarrhythmic treatment, including ICD implantation, may be warranted. PMID- 9870013 TI - Should patients with implantable cardioverter-defibrillators be allowed to drive? Observations in 291 patients from a single center over an 11-year period. AB - Motor-vehicle driving restrictions for patients with implantable cardioverter defibrillators (ICDs) vary widely throughout the world because safety concerns have never been adequately resolved in this patient population. To address this issue, we examined the driving behavior of 291 ICD patients to correlate the frequency of device therapy during driving, the occurrence of syncopal symptoms, and the incidence of traffic accidents. Fifty of the 291 patients had never driven. Of the remaining 241 patients, 171 (59%) continued driving postimplant and 70 (24%) elected to stop prior to (n = 30) or at the time of ICD implantation (n = 40). Patients were followed for a mean of 38 +/- 26 months (range < 1-124). During this period, no patients died while driving. Of 11 accidents involving 11 driving patients (6%), only 1 was caused by the driver, and none was related to syncopal symptoms or ICD therapy. Although 2 accidents (8%) occurred within 12 months postimplant, the majority (50%) took place after more than 36 months. ICD therapy was delivered in 8 patients (5%) while driving: 13% (1 episode) of the discharges occurred within the first year postimplant, 13% (1 episode) occurred between 1-2 years, and 74% (6 episodes) occurred > 2 years. None of these patients experienced syncope before or during these episodes. A multivariate analysis was unable to identify any variables that might predict increased risk of ICD therapy (with or without sudden death) while driving and consequent motor vehicle accidents. Our data suggest that such events occur only rarely. PMID- 9870015 TI - Interrelation of tissue temperature versus flow velocity in two different kinds of temperature controlled catheter radiofrequency energy applications. AB - The influence of blood flow cooling down the energy delivering electrode during temperature controlled radiofrequency energy application is an important factor for ablation success. In this experimental in-vitro study, using tempered saline as blood equivalent, we observed a highly significant increase in tissue temperature, lesion depth and required energy amount with increasing flow velocity. Second, we found significant deeper lesions with use of pulsed radiofrequency energy application compared to continuous application. We conclude that, even with lower electrode temperatures, success can be achieved dependent on the local blood flow velocity, and deeper lesions can be created with the use of pulsed radiofrequency energy application. BACKGROUND: Success in temperature controlled radiofrequency (RF) catheter ablation of arrhythmogenic areas in human hearts depend largely (among others) on the size of the electrode, developed pressure of electrode against tissue, as well as on the localization of the thermistor sensor within the electrode. In addition, the blood flow velocity at various sites of ablation is an important factor for the calculation of heat transport from the electrode, which obviously has not been given much consideration of in the past. The aim of the present in-vitro study, therefore, was to evaluate this important factor's influence on the temperature developed at the electrode and within the myocardial tissue. METHODS AND RESULTS: All experiments were carried out in a bath containing NaCl solution at 37 degrees C. Four different flow velocities were applied (0, 110, 180, 320 ml/cm2 *min). During and after temperature-controlled unipolar radiofrequency energy delivery (60 degrees C, 40 sec) the electrode temperature, the tissue temperature 5 mm in depth, and the total energy delivered were measured, as well as the actual depth of the lesion. The amount of energy applied to the electrode was regulated by the thermosensor in the electrode to obtain a maximum temperature of 60 degrees C. Two different kinds of radiofrequency energy delivery have been used: (1) continuous radiofrequency energy delivery as usual regarding clinical use, (2) pulsed radiofrequency energy delivery with a duty cycle length of 10 ms and a pause of at least the same duration during two consecutive duty cycles. At pulsed radiofrequency energy application, the energy for each duty cycle was held constant during delivery. The amount of pulses delivered to the electrode was regulated by the electrode's thermosensor. With both modes of radiofrequency energy delivery a uniform observation could be made. The more the flow velocity applied accelerated, the more the tissue temperature rose (R = 0.85; p < 0.00000001), and the lesion depth increased in spite of electrode temperature being held constant. The amount of the total energy delivered rose in proportion to the cooling down of the electrode dependent on the flow velocity (R = 0.69, p < 0.0000004). Steady-state temperatures had not been accomplished after 40 sec time. When energy was delivered at the pulsed mode, intramyocardial temperatures proved higher compared to the continuous mode with significant differences (p < 0.05) at comparable flow velocities applied between 180 and 320 ml/cm2*min and at same electrode temperatures. This resulted in significantly (p < 0.05) larger lesion depths in pulsed radiofrequency energy delivery. We suppose that this significant difference can be explained by a higher amount of total energy delivered at comparable electrode temperature in the pulsed mode as compared to the continuous mode. PMID- 9870014 TI - Electrophysiologic characteristics of different ectopic rhythms during slow pathway ablation in patients with atrioventricular nodal reentrant tachycardia. AB - The presence of ectopic rhythm has been considered to be the most important marker for successful slow pathway ablation, but the details of different ectopic rhythms have not been well described. This study included 83 consecutive patients with typical AV node reentrant tachycardia who underwent slow pathway ablation. The interval between the atrial signals of the His bundle electrogram and the distal ablation catheter [A(H)-A(Ab)], and the interval between the atrial components of the distal ablation catheter and the ostium of coronary sinus catheter [A(Ab)-A(CSos)] were measured. One hundred episodes of ectopic rhythm occurred with 81 (81%) successful applications. There are two different origins and three activation sequences of ectopic rhythms, including HIS rhythm (78 applications, the earliest atrial activation in the His bundle electrogram), CSos rhythm (6 applications, the earliest atrial signal in the coronary sinus ostium electrogram) and CSos preceding HIS (CSos-->HIS) rhythm (16 applications, the atrial activation sequences changing from CSos to HIS rhythm). The CSos rhythm had a shorter mean cycle length (445 +/- 81 vs. 511 +/- 132 vs. 579 +/- 140 ms, p < 0.05), a shorter [A(Ab)-A(CSos)] interval (-2.5 +/- 9.8 vs. 14.1 +/- 11.2 vs. 12.8 +/- 8.4 ms, p < 0.05) and a lower success rate (33% vs. 84% vs. 94% p < 0.05) than HIS rhythm and CSos-->HIS rhythm. Otherwise, the mean cycle length of ectopic rhythm was significant shorter in successful than in failed ablation (506 +/- 135 vs. 559 +/- 118 ms, p = 0.04). In conclusion, we found two different origins and three activation sequences of ectopic rhythms. CSos rhythm had a lower success rate in ablation of slow pathway, thus it was a poor marker for successful ablation. PMID- 9870016 TI - On the continuing relevance of prospective non-randomized clinical data... PMID- 9870018 TI - Intracardiac echocardiographic imaging of right atrial appendage: mass vs. pectinate muscle. PMID- 9870017 TI - Characteristics of local electrograms with diastolic potentials: identification of different components of return pathways in ventricular tachycardia. AB - BACKGROUND: Diastolic potentials are often sought as a possible site for catheter ablation in post-infarct ventricular tachycardia. However, delivery of energy at such sites is often unsuccessful. The purpose of this study was to determine the characteristics of local electrograms with diastolic potentials and to identify activation pattern which might indicate the critical portion of the return path of the ventricular tachycardia reentry circuit. METHODS: In 17 patients with post myocardial infarction ventricular tachycardia, 30 ventricular tachycardias were mapped with an 112 bipolar endocardial balloon at the time of surgery. Diastolic mapping of the return tract in ventricular tachycardia was performed. Four activation patterns were observed (15 figure 8 patterns, 2 circular patterns, 2 biregional patterns and 11 monoregional patterns). Of 3,360 local electrograms, 207 (6.2%) demonstrated a diastolic potential in ventricular tachycardia. They were classified into following four categories, based on the appearance and timing of the systolic component. Type A-1 electrogram: systolic activation was of low amplitude (< 2 mV) and was prolonged (> or = 100 msec), but preceded the onset of the surface QRS in ventricular tachycardia. Type A-2 electrogram: systolic activation was of low amplitude, was prolonged, but followed the onset of the surface QRS. Type B electrogram: systolic electrogram was fractionated, but relatively normal amplitude (2.0-3.6 mV). Type C electrogram: systolic electrogram was almost normal. RESULTS: Of all electrograms with diastolic potentials, three type A-1 electrograms (1.4%) were located at the exit of the return pathway, 11 type A-1 electrograms (5.3%) were located at the pre-exit site. No type A-1 was found at an entrance/bystander area. 21 type A-2 electrograms (10.1%) were at the pre-exit and 83 type A-2 electrograms (40.2%) were located at the entrance/bystander area, but such electrograms were never found at the exit site. 71 type B electrograms (34.3%) and 18 type C electrograms (8.7%) were located at the entrance/bystander area. To distinguish the type A-2 electrograms at the pre-exit site from those at the entrance/bystander area, the diastolic potential to QRS interval was measured. This interval at the pre-exit was significantly shorter than that at the entrance/bystander area (-47.2 +/- 10.7 vs -96.3 +/- 31.3 msec, p = 0.0001). CONCLUSION: Type A-1 electrograms indicated the exit or pre-exit site of return pathway. Type A-2 electrograms with diastolic potential to QRS interval < -50 msec indicated the pre-exit site. However, the other types of local electrograms with diastolic potential did not indicate the critical portion of the ventricular tachycardia circuit. These observations may be helpful during catheter mapping and ablation of patients with post-infarct ventricular tachycardia. CONDENSED ABSTRACT: Diastolic potentials are often sought to direct catheter ablation in post-infarct ventricular tachycardia. We investigated the characteristics of local electrograms showing diastolic activity in an attempt to determine whether critical portions of the ventricular tachycardia circuit could be identified by a typical "signature." In 17 patients with a remote myocardial infarction, 30 ventricular tachycardias were mapped with 112 bipolar endocardial balloon at the time of surgery. Diastolic potentials in association with low amplitude (< 2 mV) and prolonged (> or = 100 msec) systolic electrograms preceding the onset of QRS were found at the exit site and pre-exit site of return pathway. A similar systolic electrogram occurring after QRS onset with a diastolic potential to QRS interval of < -50 msec was found at the pre-exit site. However, other local electrograms with diastolic activity were at sites remote from the exit or pre-exit of the return pathway. These observations may be helpful during catheter mapping and ablation in patients with ventricular tachycardia. PMID- 9870019 TI - Prospective randomized trial of external versus internal transcatheter cardioversion in patients with chronic atrial fibrillation. AB - To evaluate the safety and long-term efficacy of internal transcatheter cardioversion, forty patients with chronic, lone atrial fibrillation were studied. The patients were randomised to internal transcatheter cardioversion or to conventional external cardioversion. In cases where the procedure was unsuccessful, cross-over to the alternate method was performed. Oral anticoagulation therapy was started three weeks prior to the procedure and was maintained for another three weeks following successful cardioversion. Sinus rhythm was restored in 16/18 patients (88%) in the internal cardioversion group, versus 9/22 patients (40%) in the external cardioversion group (p < 0.01). In addition, 8/13 (61%) patients who were crossed-over to internal cardioversion were successfully cardioverted to sinus rhythm. In contrast, both patients who were crossed-over to external cardioversion remained in atrial fibrillation. During a mean follow-up period of 23 months, 13 (39.3%) patients maintained sinus rhythm. Using the intention to treat principle, the recurrence rate was not statistically different between the two methods. It is concluded that internal cardioversion is more effective in acutely restoring sinus rhythm compared to external cardioversion. However, both methods have similar long-term recurrence rates. PMID- 9870021 TI - Recording of pacing stimulus artifacts by endovascular defibrillation lead systems: comparison of true and integrated bipolar circuits. AB - BACKGROUND: The occurrence ICD undersensing of ventricular fibrillation due to the presence of a pacing stimulus artifact (PSA) is in part related to the amplitude of the artifact recorded on the ICD rate sensing circuit. There is little comparative data regarding PSA amplitude recorded by commercial ICD rate sensing circuits. PURPOSE: To compare PSA amplitude recorded by commercial endovascular defibrillation leads utilizing integrated or true bipolar sensing circuits. METHODS: Nineteen large (60-120 kg) pigs were utilized. Two different commercial endovascular defibrillation leads were evaluated, each with its distal tip located at the right ventricular apex: (1) Medtronic Transvene; and (2) CPI Endotak. Three different rate-sensing circuits were evaluated: (1) Transvene true bipolar (tip-ring); (2) Transvene integrated bipolar (tip-coil); and (3) Endotak integrated bipolar (tip-coil). Using a separate pacing lead located at the left ventricular apex (n = 19 animals) or right ventricular outflow tract (n = 10 animals), pacing was performed at a pulse width of 0.5 milliseconds at outputs of 1.5, 5 and 10 volts. PSA amplitude was recorded at each output by each circuit. RESULTS: During pacing from the left ventricular apex, at each pacing output voltage the PSA amplitude recorded by the true bipolar circuit (0.6 +/- 0.1 mV at 1.5 volts, 2.0 +/- 0.5 mV at 5 volts, 3.7 +/- 0.8 mV at 10 volts) was significantly smaller than recorded by the Transvene integrated circuit (1.4 +/- 0.3 mV at 1.5 volts, 3.8 +/- 0.7 mV at 5 volts, 4.1 +/- 0.8 mV at 10 volts) or the Endotak integrated circuit (1.8 +/- 0.4 mV at 1.5 volts, 4.2 +/- 1.0 mV at 5 volts, 6.3 +/- 1.8 mV at 10 volts). During pacing from the right ventricular outflow tract, at each pacing output voltage the PSA amplitude recorded by the true bipolar circuit (0.7 +/- 0.1 mV at 1.5 volts, 1.7 +/- 0.4 mV at 5 volts, 4.0 +/- 0.7 mV at 10 volts) was significantly smaller than recorded by the Transvene integrated circuit (1.1 +/- 0.4 mV at 1.5 volts, 3.9 +/- 1.2 mV at 5 volts, 7.5 +/- 1.8 mV at 10 volts) or the Endotak integrated circuit (1.6 +/- 0.7 mV at 1.5 volts, 4.3 +/- 1.7 mV at 5 volts, 7.5 +/- 2.6 mV at 10 volts). For both pacing sites, the PSA amplitude recorded by the two integrated circuits was not significantly different. CONCLUSIONS: For a given pacing output voltage, PSA amplitude recorded by commercial endovascular rate sensing/defibrillation leads is greater when the sensing circuit is integrated than when it is true bipolar. These data may be helpful in planning ICD implantation in patients with previously implanted permanent pacemakers. PMID- 9870020 TI - The current status of single lead dual chamber sensing and pacing. AB - The use of single lead, atrial synchronous ventricular (VDD) pacing in patients with high grade atrioventricular (AV) block and normal sinus node function is an acceptable alternative to dual chamber (DDD) pacing. Implantation and follow up procedures are simplified, and cost is usually reduced by more than the cost of an additional atrial lead. With the use of either diagonally arranged dipole or closely spaced ring electrodes, reliable atrial sensing can be achieved using differential atrial amplifier and high atrial sensitivity. Also oversensing is infrequently observed using provocation tests and dynamic recordings, clinical undersensing is unusual and minimized by programming to the highest atrial sensitivity. However, as atrial pacing is not possible, loss of AV synchrony and rate response may occur for unrecognized or progressive sinus node disease and lower rate limit. The development of single lead dual chamber pacing system may overcome this limitation. Recent studies have demonstrated that atrial pacing can be effective either with the use of a special pacing lead configuration or via floating atrial electrode with a novel stimulation method. Overlapping Biphasic Impulse (OLBI) can reduce atrial pacing threshold. Early clinical experience suggested that this new pacing method can provide effective and reliable atrial pacing with a relatively low incidence of diaphragmatic pacing. Thus the problem of atrial sensing is solved with a single pass lead but further long term evaluation is required to assess the efficacy and feasibility of new instrumentation for single lead dual chamber pacing. PMID- 9870022 TI - The effect of delivered energy on defibrillation shock impedance. AB - The impedance of internal defibrillator shocks is an important determinant of defibrillation efficacy. To assess the effect of delivered energy on impedance, we studied 97 patients with 4 different lead systems. The lead systems evaluated were two epicardial patches, a hybrid system of a patch and right atrial coil, a dual coil transvenous lead and a transvenous lead with a subcutaneous patch. Impedances were measured for 6 shock energies between 0.1 and 30 J. Shock impedance increased at low energies for all lead systems (p < 0.001), although the rate of increase varied markedly between systems. The energy factor (FE), which is the ratio of impedances for the 0.1 and 10 J shocks, was least for the platinum transvenous lead (1.2 +/- 0.02) and greatest for the titanium hybrid lead (4.2 +/- 0.2). Reversing the polarity of the hybrid lead markedly attenuated the impedance rise. These findings indicate that there is at least a modest rise (20%) of shock impedance at very low delivered energies. The largest increases noted with titanium lead systems are primarily due to polarization. Titanium transvenous leads should be avoided when low energy shocks are utilized such as for the cardioversion of ventricular tachycardia or atrial fibrillation. PMID- 9870023 TI - Effect of catheter tip length and position on lesion volume in temperature controlled RF ablation in canine tricuspid valve annulus. AB - BACKGROUND: Radiofrequency (RF) energy has been delivered to the tricuspid valve annulus (TVA) in humans with both 4 mm and 8 mm long catheter tip electrodes to treat atrial flutter. However, lesion volume with temperature controlled RF delivery systems has not been previously characterized. METHODS: In 10 anesthetized canines, a single pulse of temperature controlled RF energy at a 70 degrees C set point, 30 second duration was delivered with either a 7 Fr/4 mm tip or a 7 Fr/8 mm tip electrode in a position both anterolateral and posteroseptal to the tricuspid valve annulus (TVA). Surface echocardiogram was obtained prior and after ablation. The animals were sacrificed after ablation and the lesions underwent gross and histological examination. RESULTS: Lesion size, tip temperature and power were related to tip electrode surface area (SA). Eight mm tips (SA = 59 mm2) tended to create significantly larger lesions than 4 mm tips (SA = 29 mm2). Median lesion volume was 22 vs. 1.5 mm3, respectively. Eight mm tips were also associated with higher power requirements and lower temperatures than 4 mm tips. Posteroseptal TVA lesions tended to be larger than anterolateral lesions. No significant complications were noted. CONCLUSIONS: Using temperature controlled RF ablation, large lesions may be safely created on the canine TVA using 7 Fr catheters with 8 mm long tips. PMID- 9870025 TI - Radiofrequency catheter ablation of two right Mahaim-like accessory pathways in a patient with Ebstein's anomaly. AB - A 17-year-old woman with Ebstein's anomaly and recurrent episodes of antidromic tachycardia with two distinct morphologies is described. The tachycardias were produced by two separate Mahaim-like accessory pathways. These were localized by their activation potentials at the anterolateral ventricular margin of the tricuspid annulus and ablated in a single session using radiofrequency current. PMID- 9870024 TI - Feasibility of cardiac cryoablation using a transvenous steerable electrode catheter. AB - We investigated the feasibility of using cryogenic technology in an electrode catheter for percutaneous ablation of cardiac tissue. Despite its high success rate, radiofrequency catheter ablation has important limitations especially with regards to the treatment of ventricular arrhythmias associated with a chronic scar. Arrhythmia surgery experience has shown that freezing with a hand held probe can permanently ablate the arrhythmogenic substrate of ventricular tachycardia associated with an old scar. Moreover, cryosurgery also allows for reversible "ice mapping," in which the area likely responsible for the arrhythmia can be evaluated by suppressing its electrophysiologic properties prior to the creation of an irreversible state. A new steerable cryoablation catheter using Halocarbon 502 as a refrigerant was utilized in six dogs. Serial cryoapplications were performed in the right and left ventricles. In two dogs, we attempted reversible ice mapping of the AV node. Pathological evaluation of the lesions was done acutely in all the animals. Forty-two cryoapplications were delivered at a mean temperature of -45 +/- 9.8 degrees C. No lesion was found at pathological evaluation for 16 cryoapplications which did not achieve a temperature of less (colder) than -30 degrees C. The remaining applications resulted in 26 lesions which were hemorrhagic and sharply demarcated from normal myocardium. Histological evaluation revealed contraction band necrosis. Reversible ice mapping of the AV node was successfully achieved in two animals. Cryoablation is feasible using an electrode catheter with multiple electrodes. This technology has the potential to allow for reversible ice mapping to confirm a successful ablation target before definitive ablation. PMID- 9870026 TI - Catheter ablation of Wolff-Parkinson-White syndrome associated with congenital absence of inferior vena cava. AB - In the present report we describe a patient (a 36-year-old woman with 15 year history of supraventricular tachyarrhythmias) with congenital absence of inferior vena cava (IVC) revealed during radiofrequency (RF) catheter ablation procedure for right postero-septal Wolff-Parkinson-White syndrome (WPW). For the absence of IVC, the ablation procedure was more difficult, because we had to perform the ablation with the catheters (the ablator catheter and the coronary sinus catheter) introduced both through the superior vena cava. The application of RF energy (35 Watt for 60 seconds) at successful site abolished accessory pathway conduction. The following day was performed the venous angiography, showing the absence of the IVC and a venous return via paravertebral venous plexus to the azygous vein and superior vena cava into the right atrium. Computer tomography confirmed the absence of the IVC with azygous continuation. The drainage via the azygous system modified the radiological image on chest roentgenogram of the right mediastinal silhouette. During cardiogenesis fusion of the IVC and organisation of the heart occur between the 33rd to 40th embryonic days. It is therefore possible that some unknown teratogenic mechanism at this critical period might have caused, in the patient, both the developmental arrest of IVC and failure of regression of atrio-ventricular anatomical and electrical continuity in the right postero-septal region. PMID- 9870027 TI - Transient complete atrioventricular block following radiofrequency ablation of left free wall accessory pathway. AB - We report the case of a forty-six-year-old female with symptomatic WPW syndrome. The accessory pathway was located on the left free wall, for which ablation was attempted from the retrograde aortic approach. The ablation catheter was positioned at the appropriate site on the mitral anulus. A single radiofrequency energy application resulted in complete AV block with no escape rhythm, necessitating ventricular pacing. The AV conduction soon resumed with no evidence of pre-excitation. This phenomenon was thought to be related to trauma to the AV node during catheter entry in to the left ventricle. PMID- 9870028 TI - Introduction: progress and enigmas in cancer epidemiology. PMID- 9870029 TI - The global burden of cancer. AB - The number of new cancer cases, and the annual incidence rates, of 25 different cancers have been estimated for the year 1990 for every country of the world. The distributions of the most common cancers in men and women are presented for 23 broad 'Areas'. The total number (excluding non-melanoma skin cancer) was 8.1 million, just over half of which occur in the developing countries. The most common cancer is lung cancer, which accounts for 18% of cancers of men worldwide. Stomach cancer is second in frequency (almost 10% of all new cancers) and breast cancer--by far the most important cancer of women (21% of the total) is third. There are very large differences in the relative importance of the different cancers by world area; some of the factors, environmental and genetic, underlying the geographic distributions, are discussed. PMID- 9870030 TI - Growing evidence that several human cancers may originate in utero. AB - Hormone-related cancers may originate in utero. Accumulating evidence is given by animal studies, descriptive epidemiologic data and analytic etiologic studies. Indicators of high levels of endogenous pregnancy hormones, like high birth weight and jaundice in the offspring, are associated with increased risk for breast, prostate, and non-seminoma testicular cancer. Indicators of low levels, like pre-eclampsia, are associated with decreased risk. These results support the hypothesis that the pre- and perinatal periods constitute a 'time window' that is of major importance for future cancer risk. PMID- 9870031 TI - Dietary fat intake and cancer risk: a controversial and instructive story. AB - Strong correlations among countries between per capita dietary fat consumption and rates of cancers of the breast, colon, and rectum have suggested possible causal relationships. However, in large prospective studies, total fat intake has not been associated with risk of breast cancer, suggesting that the international correlations are seriously confounded by differences in other variables related to affluence, including reproductive variables, physical activity, and food availability. For colon and prostate cancer, the data from prospective studies are more limited, but also suggest that fat per se is not a risk factor. High red meat consumption may be related to risk of colon cancer independent of total fat intake, and some factor associated with consumption of animal fat, but not vegetable fat, appears to be related to risk of prostate cancer. Excess body fat, which is determined by the balance between physical activity and energy intake from all sources, is an important cause of post-menopausal breast cancer and probably colon cancer. These developments suggest that international comparisons of diets and disease rates should be interpreted cautiously and may be highly misleading. PMID- 9870032 TI - Towards an understanding of breast cancer etiology. AB - We present an etiological model for breast cancer in humans, and we examine whether it accommodates the patterns of occurrence of this disease and the associated risk factors. The model has four components: (1) the likelihood of breast cancer occurrence depends on the number of cells at risk; (2) the number of target cells is partially determined early in life, perhaps even in utero; (3) while a pregnancy stimulates the replication of already initiated cells, it conveys long-term protection through structural changes, terminal cellular differentiation, and perhaps other mechanisms; and (4) in adult life, mammotropic hormones, in conjunction with their receptors, affect the number of target cells, the likelihood of retention of spontaneous somatic mutations, and the rate of expansion of initiated clones. The model accommodates several hypotheses but also allows new insights. PMID- 9870033 TI - What causes prostate cancer? A brief summary of the epidemiology. AB - In recent decades, prostate cancer has emerged as one of the most common diseases among older men, particularly in Western society. Several years ago, only age, race, and family history were known risk factors for this disease. However, today, much progress has been made towards discovering nutritional and hormonal risk factors for prostate cancer. Biomarkers, including testosterone and insulin like growth factor, and nutritional factors, especially meat, fat, and dairy intake, have been linked to greater risk of disease. Higher consumption of selenium and vitamin E, fructose/fruits, and tomatoes all have been associated with reduced occurrence of prostate cancer, but as yet their efficacy for prevention remains unproven. The challenge of understanding the enigmas of this disease will continue into future decades, as we convert current knowledge into preventive and therapeutic recommendations. PMID- 9870034 TI - Is Helicobacter pylori really the cause of gastric cancer? AB - Since 1994, when the International Agency for Research on Cancer (IARC) designated Helicobacter pylori a human carcinogen, a considerable number of new studies have examined the association of this microorganism with risk of gastric cancer. The aim of this review was to critically evaluate current evidence of a causal relationship between the infection and gastric cancer in humans, taking these new studies into account. A MEDLINE search for the years 1982 through March 1998 yielded eight cohort studies, 40 case-control studies and eight ecologic studies. They were divided into four categories according to the weight of the evidence produced. In the category producing the strongest evidence, the cohort studies, five out of eight studies (including the three that formed the basis for the IARC decision) found a statistically significant excess risk of gastric cancer among the infected with odds ratios ranging between 2.8 and 6.0. Two cohort studies showed a non-significant tendency towards a positive association, but with point estimates indicating no more than a 50-60% excess risk among the H. pylori seropositive, and one apparently well-performed cohort study was completely negative. The two population-based case-control studies published to date found statistically non-significant odds ratios no higher than 1.3 and 1.4. Thus, the evidence of a strong causal link between H. pylori infection and gastric cancer has, if anything, become less convincing. Recent studies seem to indicate that the strength of the association may vary with type of H. pylori strain, and possibly presence of effect-modifying co-factors in the host and/or the environment. Future studies need to identify high-risk constellations of carcinogenic strains and unfavorable co-factors to make targeted prophylaxis cost effective. PMID- 9870035 TI - Genes and environment in the etiology of colorectal cancer. AB - Environmental factors, including diet, are important in the etiology of colorectal cancer. Genetic pathways of hereditary and sporadic bowel cancer are being elucidated. Highly penetrant mutations in certain genes, such as mismatch repair genes, play a major role in development of hereditary colorectal cancer, while interactions between low penetrance polymorphisms in carcinogen metabolizing enzymes and environmental factors may identify susceptible subgroups and shed light on the etiology of sporadic colorectal cancers. In this review we summarize the current state of knowledge regarding the role of gene-environment interactions in development of colorectal cancer and discuss areas of particular promise, as well the limitations of these studies. PMID- 9870036 TI - Towards global control of liver cancer? AB - On a global scale, liver cancer is one of the leading causes of cancer morbidity and mortality. However, liver cancer rates vary substantially by country, with more than 80% of liver cancer cases occurring in the developing world. The major risk factors for hepatocellular carcinoma (HCC), the predominant histologic sub type, are considered to be chronic infection with hepatitis B virus (HBV), hepatitis C virus (HCV) infection, exposure to aflatoxins, and excessive alcohol consumption; tobacco smoking and oral contraceptive use also may be associated with increased risk of HCC. This paper focuses on those risk factors that represent new targets for intervention, namely HBV and HCV infections and aflatoxin exposure. Childhood vaccination against HBV represents the greatest potential for reducing the liver cancer burden and could lead to the elimination of as much as 60% of all cases. PMID- 9870037 TI - The role of human papillomaviruses in anogenital cancers. AB - There is substantial evidence to suggest that common risk factors exist for cancer of the cervix and other anogenital cancers. Cervical cancer has been etiologically linked with venereal types of human papillomavirus (HPV) and the same types of HPV are being found in other anogenital tissues. The question is to what extent these HPV types may also contribute to carcinogenesis in anogenital cancer other than cervical cancer. In this review some general aspects of the natural history of HPV are presented followed by a description of the incidence and risk factors for anal, vulvar, and penile cancer. Main focus is given to the evidence for an association between these cancers and HPV. PMID- 9870039 TI - Wandering behavior and attempted exits among residents diagnosed with dementia related illnesses: a qualitative approach. AB - Exiting attempts and wandering behavior among residents diagnosed with dementia related illnesses were investigated. Residents were exposed to three test conditions: a mini-blind that concealed the view from the door, a cloth panel that concealed the panic bar of the door, and both the mini-blind and the cloth panel. During these test conditions, qualitative observations were conducted in addition to recording the attempted exits. The purpose of this paper is to present the qualitative findings in relationship to a model developed. The findings suggests that residents seemed to exit due to goal-orientation, exit seeking behaviors, and attraction to the panic bar. PMID- 9870038 TI - The contribution of small life events to the psychological distress of married and widowed older women. AB - The contribution of small life events to the prediction of general psychological distress was examined for 50 married and 21 recently widowed older women. These two groups were contrasted as having or not having experienced an uncontrollable major life stressor (i.e., the recent death of a spouse). Negative small life events (i.e., daily hassles) contributed above and beyond general demographic factors; conjugal bereavement status; social support; other, non-conjugal bereavement, major life events; and the interaction of these life events and social support in the prediction of general psychological distress. Results support assessing negative small life events as well as major life events for both married and recently widowed older women. PMID- 9870040 TI - The relationship of reciprocity to self health care in older women. AB - According to Social Exchange Theory, reciprocity in relationships is essential to functioning within the social context and is an important component in well-being as people age. Because of a potential relationship to meaning in life and hence to self health behaviors this study addressed: (1) Does reciprocity relate to self health care and (2) does reciprocity contribute to the explanation of self health care within a model including functional status, satisfaction with social support, and sense of meaning in life? Data were collected from 69 community dwelling women aged 65 to 99. Analysis indicated general health perception, support provided to others and sense of meaning in life were significant predictors of self health care. PMID- 9870041 TI - Daughters' caregiving decisions: from an impulse to a balancing point of care. AB - Daughters comprise 29% of all caregivers of the dependent elderly and make decisions about parent care in a context of ambiguous and changing role expectations. This qualitative study explored the decision process of ten caregiving daughters. The women responded to parent dependency with an impulse to care and pursued a balancing point of care in the face of limiting decision conditions. Each daughter constructed a sense of "enough": an equation of her multiply-determined impulse to care and her personal threshold of support. These constructions help explain the variability of levels, lengths, and forms of parent care. PMID- 9870042 TI - Women's narratives: evolving symptoms of myocardial infarction. AB - Diagnosing acute myocardial infarction (AMI) in women is difficult since chest pain may not be a hallmark symptom. Research is needed to identify symptoms experienced by women with AMI to facilitate timely diagnosis. The purpose of this study was to identify new symptoms and their evolution experienced by women prior to diagnosis of AMI. Non-probability sampling was used to select 20 diverse women. Intensive home interviews were conducted and transcribed. Content analysis and constant comparison were used to develop nine data clusters: Location of Pain; Intensity of Pain/Sensations; Cardiovascular/Temperature Changes; Respiratory Sensations; Gastrointestinal Symptoms; Emotions; Hand and Arm Sensations; Neurological/Vision Changes; and Fatigue. Some women progressed to AMI in minutes while others had symptoms for weeks. Findings should increase awareness of women's symptoms of AMI. Further research is needed with a larger sample. PMID- 9870043 TI - Spirituality in the lives of older women. AB - Little is known about the importance of spirituality in the older women. This study explores the role of spiritual in the lives of women 70 years of age and older. A convenience sample of 56 respondents responded to the JAREL Spiritual Well-Being Scale and a demographic data questionnaire. Using an analysis of variance, significance was found for those who did not attend school beyond high school and those who did. There were difference between Catholics and Protestants on the Faith Belief Dimension factor and the total spirituality score. PMID- 9870044 TI - Rural-urban women's experience of symptoms of depression related to economic hardship. AB - This study compares rural and urban women's experiences of depressive symptoms. Cross-sectional data were used from 623 women 55 years of age and older who were part of a randomly selected community based sample. Theoretical perspectives included individual stress and community context as explanatory factors contributing to differences in the proportion of women who identify having had the experience of symptoms of depression. Results of the analysis using correlation and multiple regression supported the importance of perceptions of financial status. Community type and age were not related to the level of depressive symptoms reported by the women who were interviewed. PMID- 9870046 TI - The impact of employment and serious illness on grandmothers who are raising their grandchildren. AB - Increasing numbers of children are living in families consisting of grandparents and grandchildren. This paper presents findings of a study in which 123 caregiving grandmothers were interviewed. Fifty-eight percent of these grandmothers were "career caregivers," whose homes were always filled with a child or grandchild. Most of them were in good to excellent physical health and their mental health was at least as good as that of a national sample. Regression analyses found that having a life-threatening physical condition, being younger and white explained psychological anxiety; and having a life-threatening physical condition and not being employed explained psychophysiological mental health symptoms. These findings suggest that some grandmothers are at risk for mental health symptoms and raise questions about the implications of caregiving of grandchildren for women. PMID- 9870045 TI - Social physique anxiety in postmenopausal women. AB - Social physique anxiety (SPA), the degree to which individuals become anxious when others observe their bodies, is an important concept for postmenopausal women because it may be directly related to their physical activity behavior. Women with high levels of social physique anxiety may not participate in health enhancing physical activity. Therefore, the main purpose of this study was to examine SPA in postmenopausal women relative to leisure time physical activity (LTPA, activities such as exercise, recreation, and sports). Other factors, such as percent body fat, body fat distribution, age, and hormone replacement therapy status were also examined for their relationship to SPA. Women who expended < or = 500 kcal.wk-1 in LTPA had significantly higher SPA than women who expended > or = 2,000 kcal.wk-1, independent of percent body fat. Women with more than 37.5% body fat had significantly higher SPA than those with less than 37.5% body fat. Women with upper BFD (waist-to-hip ratio or WHR > 0.85) had higher SPA than women with lower BFD (WHR < 0.75). There were no differences in SPA relative to age or HRT status. It appears that a sedentary lifestyle, high percent body fat, and upper body fat distribution are associated with increased SPA in postmenopausal women, thus health promotion professionals should be aware of these concerns when developing physical activity interventions for postmenopausal women. PMID- 9870047 TI - Sex comparisons in life satisfaction and psychosocial adjustment scores with an older adult sample: examining the effect of sex role differences in older cohorts. AB - The present study examined how sex and other individual-difference factors (i.e., age level, locus of control orientation, and self-actualization subscale scores) relate to older adults' scores on life satisfaction and psychosocial adjustment. Seventy-eight older adults (n = 39 females) were recruited from independent living retirement communities located in Pennsylvania. Results indicated that females in the sample were not significantly different in mean life satisfaction scores but were significantly lower in mean psychosocial adjustment scores than males in the sample. There were no significant age-level differences in mean scores. Qualitative data from unstructured post-testing interviews revealed that women were more likely to express regret and sometimes frustration toward perceived "missed opportunities" in life (e.g., career) due to expected social roles of being a wife and mother in the decades ranging from the 1920s through the 1960s; these feelings of regret or frustration were not expressed by any of the males in the study. PMID- 9870048 TI - Predictors for osteoporosis diagnosis among postmenopausal women: results from a national survey. AB - Osteoporosis is a serious health problem in the U.S. today resulting in premature mortality, deformity, pain, loss of function, and disability. Diagnosis of osteoporosis is key to obtaining treatment. The purpose of this study was to examine risk factors that are correlated with osteoporosis diagnosis among a national sample of postmenopausal women. Factors associated with osteoporosis diagnosis included age, race, and family history. Health care providers, including physicians, dietitians, nurses, physical therapists and social workers, can play a key role in improving diagnoses among at-risk women. They can work together to increase awareness and improve knowledge regarding the risk factors for osteoporosis including body mass index, physical activity, smoking and nutritional status. PMID- 9870049 TI - Gender role orientation as a correlate of perceived health, health behavior, and qualitative well-being in older women. AB - This study investigated the relationships of gender role orientations (Feminine, Masculine, & Androgynous) with self-perceived health status, health behavior, and qualitative well-being experience in a simple random sample of community-dwelling older women (N = 70). Instruments used were The Bem Sex Role Inventory (Bem, 1974); The Seniors' Lifestyle Inventory (Schwirian, 1991); and The Integration Inventory (Ruffing-Rahal, 1991). Findings revealed the greatest number of significant positive correlations with the Feminine orientation; Androgyny was significantly linked with qualitative well-being and with a positive self comparison in relation to peers and Masculinity was positively linked with health behavior. Findings are interpreted in light of successful aging issues for older women. PMID- 9870050 TI - Gender differences in coping and control with memory aging. AB - This secondary analysis of data from a large study of memory perceptions among the elderly examined gender differences in control, coping, health, and metamemory and explored the influence of these factors on anxiety and depression in the elderly. Adults 55 years of age and older, 128 female and 41 males, were recruited from continuing education programs in two Southern states. Females were older than males and reported that their memories were better overall than males. There were no differences between the groups in depression, health, or memory control variables. Females had significantly greater state anxiety than males but no differences were seen in domain-specific memory anxiety or other metamemory domains. Females scored higher than males on help-seeking, existential growth, religiosity, and total coping strategies. In the two regression models the set of study variables predicted 79% of the variance in depression and 15% of the variance in memory anxiety. The addition of perceived health status to each model substantially changed each of their predictive values. PMID- 9870051 TI - Women, marital status, and symptoms of depression in a midlife national sample. AB - Previous studies of the correlates of depression among women have not generally been based on adequate midlife samples or precision in the specification of marital status categories. The present analysis is designed to address these deficiencies and is based on data from the Health and Retirement Survey-Wave 1 (i.e., respondents 51 to 61 years old). Results indicate that married women are less likely to report symptoms of depression than their unmarried counterparts. The mental health benefits of marriage are greater for men than for women. Moreover, other variables, such as marital satisfaction, self-rated health, and employment status are more powerful predictors of emotional well-being in midlife than marital status per se. The quality of marriage affects depressive symptoms more strongly for women than men. PMID- 9870052 TI - Longitudinal employment decisions of daughters and daughters-in-law after assuming parent care. AB - Employment decisions of full-time, part-time and not employed daughters and daughters-in-law (n = 157) are described over the first 18 months of parent care at two points in time: three months after assuming care and 15 months later. Employment decisions were also examined when caregiving continued (n = 55), when it was discontinued (n = 55) and when a parent died (n = 47). Findings indicate that part-time employed daughters make more changes and adjustments in their employment than do full-time or not employed daughters. When a parent died, more changes in employment occurred prior to the death than when a parent remained in care. PMID- 9870053 TI - Thriving older African American women: aging after Jim Crow. AB - This paper is based on the findings of small group discussions with self-defined successful African American women age 60+ in Charlotte, North Carolina. These women, who lived through the Jim Crow era and thrived in spite of the obstacles, continue to seek meaning in their lives through the roles they play in their families, churches, and communities. They feel strongly that there is a core of key values that continue to hold meaning and struggle to impart these values to those whose lives they touch. The key values identified include education, religion, work, and giving back to the community, and illustrate the integration of both traditional and nontraditional definitions of success. Mentoring is proposed as an important concept for understanding the lives of African American women in later life. PMID- 9870054 TI - The costs of radiosurgical treatment: comparison between gamma knife and linear accelerator. AB - Radiosurgical treatment can be carried out by means of a Gamma Knife or a Linear Accelerator. The Linear Accelerator may be either a single-purpose appliance, exclusively employed in radiosurgery, or an adapted appliance, which is used primarily for fractioned radiotherapy, and only additionally for radiosurgical purposes. The first alternative will be referred to briefly as a "dedicated Linac", the latter as an "adapted Linac". Cost accounting data for these alternatives will be discussed under three main categories: investment costs, operating costs, and finally staffing costs. Costs are only considered to the extent that this is necessary to facilitate a comprehensive cost comparison. Factors for which the costs remain the same or at least broadly the same will from the outset not be taken into consideration. These include, for instance, the costs of general or special administration, diagnosis, and image processing. The results and conclusions of this study therefore cannot be employed immediately in the evaluation of cost reimbursement schemes of the type carried out by agencies responsible for social insurance. Here, appropriate complete cost analyses especially for this purpose are required. The final comprehensive cost comparison reveals that the adapted Linac is the most favourable alternative with small annual quantities of patients. With larger numbers of patients the Gamma Knife represents the most favourable from a cost accounting angle. The dedicated Linac accordingly does not have a cost advantage for any of the examined numbers of patients. Clearly the lowest treatment costs per patient can be achieved by employing a Gamma Knife and using it to capacity. PMID- 9870055 TI - The Camino intracranial pressure device in clinical practice: reliability, handling characteristics and complications. AB - Intracranial pressure monitoring has a key role in the management of patients developing increased intracranial pressure (ICP). We adopted the Camino fiberoptic system for intracranial pressure measurement in 1993 in our neurosurgical department. The aim of this study was to investigate reliability, handling characteristics and complication rate of the Camino intracranial pressure device. In an eighteen month period, we prospectively investigated 118 patients with intracranial pathology undergoing Camino fiberoptic intraparenchymal or intraventricular ICP monitoring. The assessment of reliability of ICP monitoring according to patients clinical condition, to cranial computed tomography (CCT) findings and ICP waveform was carried out. Position of the probe and intracranial bleeding complications related to probe insertion were confirmed by CCT. Technical complications, as well as infections due to the device, were documented. In vivo recalibration was performed in 22 patients. At the end of the measuring period the drift of the probe was evaluated and the accuracy of the fiberoptic device was measured by performing a two point calibration. Recordings of intracranial pressure were carried out with 136 Camino devices (104 parenchymal, 32 ventricular) in 118 patients with an average measuring time of 94.1 +/- 79.1 hrs. One hundred and fifteen Camino intracranial pressure devices (85.2%) demonstrated reliability according to the predetermined clinical parameters. The actual mean drift after removal of the devices was 3.4 mmHg +/- 3.2 with an actual daily drift of 3.2 +/- 17.2 mmHg. Recorded complications included infection (0.7%), intraparenchymal haematoma (5.1%), and a high complication rate (23.5%) with regard to technical aspects. The Camino intracranial pressure system offers reliable ICP measurements in an acceptable percentage of devices, and the advantage of in vivo recalibration. The high incidence of technical complications identifies a need for improvement in the fiberoptic cable and the fixation system. PMID- 9870056 TI - Factors affecting the outcome of foetal hydrocephaly. AB - In this study, the authors attempt to provide an account of the factors that affect the outcome of hydrocephaly in 26 foetuses. The hydrocephalus was related to a myelomeningocele in 35% of patients. Sixty-two percent of foetuses showed intra-uterine progression of their hydrocephalus and 50% were shunted postnatally. At a mean follow up of 2 years, the outcome was considered "fair" in 54% of patients. Our findings demonstrate that in foetal hydrocephaly a more favourable outcome is expected in patients with hydrocephalus which does not progress in utero, in whom the labour is not induced before 36 weeks of gestation, who are delivered vaginally weighing more than 2.5 kg and have a head circumference below the 95th centile and a CT cortical mantle thickness of 2 cm and more and who are treated by CSF shunting. The diagnosis of the foetal hydrocephaly at or before 28 weeks of gestation and the presence of a myelomeningocele did not affect the outcome significantly. Neurosurgeons are reminded to keep an open mind for infants with foetal hydrocephaly and to offer active treatment to patients with a potentially favourable outcome. PMID- 9870057 TI - Four-year experience with the routine use of the programmable Hakim valve in the management of children with hydrocephalus. AB - OBJECTIVE: Cerebrospinal fluid (CSF) over- and underdrainage symptoms are frequent sequelae of shunt placement in patients with hydrocephalus, sometimes requiring repeated operations. To achieve more adequate CSF drainage, the non invasively programmable Hakim valve has been developed. Because the clinical experiences with this valve so far are confined to adults, we describe our experiences with the routine use of the programmable Hakim valve in childhood hydrocephalus. METHOD: Sixty children (mean age of 3.4 years) with hydrocephalus of various aetiologies have been shunted with the programmable Hakim valve. In the majority of cases, initial opening pressures of between 100 and 120 mm H2O were selected. The mean follow-up period was 2.1 years. RESULTS: Thirty-three readjustment of the pressure setting of the valve were performed in 20 children because of CSF overdrainage (low intracranial pressure syndrome n = 13, slit ventricle syndrome n = 2, hygroma n = 1), CSF underdrainage (n = 3) and CSF leakage through the operation wound (n = 1). The symptoms of inadequate CSF drainage were cured in 18 of the 20 children. The necessity for valve readjustments was independent of the aetiology of the hydrocephalus. Thirty-one complications requiring repeated operation occurred during the follow-up period, accounting for an annual complication rate of 24.6%. Three complications were valve-related. CONCLUSION: In the majority of cases, the programmable Hakim valve allows the successful management of symptoms related to CSF over- and underdrainage by non-invasive change of the initial pressure setting of the valve. Therefore, the programmable Hakim valve should be considered as an alternative to non-programmable valves of advanced design. PMID- 9870058 TI - Effect of hyperosmotic solutions on human brain tumour vasculature. AB - Reversible opening of the blood-brain barrier (BBB) has been used to increase delivery of chemotherapeutic agents into brain tumours, but it is complicated and requires general anaesthesia. Without affecting the normal BBB, and avoiding the complications of BBB modification by hyperosmotic solution, we tried an adequate minimal BBB disruption in brain tumours. Although the effect of BBB disruption on normal brain has been described, there are no reports of the effect of an impaired BBB on microcirculation. In this study, four patients underwent surgical resection of a glioblastoma multiforme (GM; n = 1), astrocytoma (n = 2), or metastatic brain tumour (n = 1). Epicerebral microcirculation was observed in the operative field. Serial fluorescein microangiograms of the tumour and peritumoural area were obtained before and after BBB disruption was introduced intra-operatively by retrograde infusion of mannitol introducing a catheter via the temporal superficial artery back to the carotid bifurcation. On the initial microangiogram, staining by the fluorescein dye was observed in the GM and metastatic tumour but not in the astrocytoma; no extravasation of fluorescein dye was observed in the peritumoural areas. After BBB disruption, fluorescein perfusion increased and extravasation of fluorescein dye from the venules was observed in the GM and the metastatic tumour and in the peritumoural area of both lesions; BBB disruption started from venules in the peritumoural area without affecting the normal brain. However, such effects were not observed in the astrocytomas after BBB disruption nor in normal brain tissue in any patient. It appears that the integrity of the BBB is less stable in the peritumoural area of GM and metastatic brain tumours than it is in astrocytomas or normal brain. Osmotic BBB disruption may offer a method for achieving global delivery of therapeutic agents to brain tumours and peritumoural areas. PMID- 9870059 TI - Prediction of facial nerve displacement in extra large vestibular schwannoma. AB - The primary objective in the surgery of extra large vestibular schwannoma is the total removal of the tumour mass while preserving the facial nerve. Preservation of the facial nerve in extra large tumours is reported as being notoriously difficult in the majority of cases. This study was undertaken to evaluate the accuracy in predicting displacement of the facial nerve by preoperative radiological imaging studies in 19 cases of extra large vestibular schwannoma. The direction of displacement of the facial nerve was predicted with preoperative axial and coronal MRI scans and verified intraoperatively. We achieved total removal of tumours in 84.2%, facial nerve displacement was predicted in 80% and we accomplished anatomical preservation in 80%. Prediction of displacement was difficult in tumours with little or no intracanalicular components or with severe bony destruction of the internal acoustic meatus. The preoperative prediction of facial nerve displacement in extra large tumours allows safe internal decompression of the tumour and careful dissection near the predicted area of the facial nerve during the operation. Consequently, a high rate of anatomical preservation of the facial nerve can be achieved. PMID- 9870060 TI - Oculomotor nerve neurinoma: report of two cases. AB - Being motor nerves neurinomas originating from ocular nerves are very rare, unless associated with neurofibromatosis. Authors describe two cases of oculomotor nerve i.e. third nerve, Schwann cell tumours. One of them presented as a cavernous sinus mass in a middle aged lady while the other was a middle aged man with a large cisterno-cavernous tumour. Surgical approach is discussed and the relevant literature reviewed. PMID- 9870062 TI - The value of intraoperative angiography for surgical treatment of cerebral arteriovenous malformations in eloquent brain areas. AB - Intraoperative digital subtraction angiography (DSA) allows intraoperative assessment of outcome of cerebral arteriovenous malformations (AVM). This study reports on 21 patients with AVMs in eloquent areas of the brain extirpated between July 1995 to March 1998. Extirpation was always followed by intraoperative DSA. Intraoperative angiography disclosed an occult residual nidus in 4 cases (19%). Complete extirpation of the AVM was achieved in all cases. Following surgery the neurological condition improved in 15 cases (71%), remained unchanged in 5 (24%), and worsened in 1. There were no secondary postoperative haemorrhages, nor complications related to the angiography. These results indicate that intraoperative DSA should be considered in the course of surgical treatment of cerebral AVMs in eloquent areas of the brain. PMID- 9870061 TI - Aneurysmal remnants after microsurgical clipping: classification and results from a prospective angiographic study (in a consecutive series of 305 operated intracranial aneurysms). AB - The aim of this prospective study, carried out in a consecutive series of 305 microsurgically clipped aneurysms, was to check the absence of an aneurysmal remnant on post-operative angiography, and if a remnant was found to quantify its size in order to consider additional clipping to avoid the risk of rebleeding. Out of the 305 aneurysms, 292 (96%) were located in the anterior and 13 (4%) in the posterior circulation. Post-operative angiography was performed on average two weeks after surgery. Determination of the presence or not of an aneurysmal remnant and its quantification was done by an independent observer (JCA). Aneurysmal remnants were classified into 5 grades: grade I: less than 50% of neck size, grade II: more than 50% of neck size, grade III: residual lobe of a multilobulated sac, grade IV: residual sac of less than 75% of aneurysmal size and grade V: residual sac of more than 75% of aneurysmal size. Correlations between presence (and size) of the remnant and anatomical-surgical data obtained from the operative report were studied. Clipping was considered incomplete in 18 of the 305 aneurysms (5.9%). The group with residual neck only (grade I = 8 cases, Grade II = 4 cases) amounted to 4% of the whole series, whereas the group with residual neck + sac (grade III = 4, grade IV = 1, Grade V = 1) to 1.9%. Only this latter group was amenable to re-operation for complementary clipping without creating a stenosis of the parent artery. Our results are in the range of those of other published series. Anatomical-surgical factors for predisposition to incomplete clipping are discussed. The rates of sac obliteration using microsurgical clipping are to be compared with those recently achieved by electrically detachable coiling. The classification which we have developed is proposed for future comparison with endovascular results. PMID- 9870063 TI - Wrapping of solitary ruptured intracranial aneurysms, outcome at five years. AB - Previous follow up studies of patients who have had ruptured intracranial aneurysms treated by a wrapping technique have largely been in the pre microsurgical era. Our objective was to ascertain whether wrapping aneurysms with the aid of the operating microscope provides protection against rebleeding in the short and long term. The study involved retrospective analysis of patients with ruptured aneurysms treated by wrapping over a six-year period. Twenty-nine patients were identified with wrapped aneurysms, of these 15 had wrapping of a solitary ruptured aneurysm, the remainder were excluded because of clip/wrap combination, multiple aneurysms, and unruptured aneurysms. Of the 15 patients who had a solitary ruptured aneurysm wrapped, none had rebled at one year follow up. One patient died of a myocardial infarction at 4 years, the remaining 14 patients had no episodes of rebleeding with all patients achieving Glasgow outcome scores of four or five at 5 years follow-up. Although surgical clipping of intracranial aneurysms is the definitive method of treatment, our findings suggest that wrapping of ruptured intracranial aneurysms with the aid of the operating microscope confers good protection against both early and late rebleeding in those cases considered 'unclippable'. PMID- 9870064 TI - Atypical hemifacial spasm. AB - Among 155 cases of hemifacial spasm (HFS), the authors found two cases of atypical HFS (1.3%) in which spasm started with the orbicularis oris and buccinator muscles, and gradually spread upward to involve the orbicularis occuli muscle, whereas the reverse process is usually seen in cases of typical HFS. The compression site in cases of atypical HFS is the posterior/rostral aspect of the facial nerve (FN), whereas it was the anterior/caudal aspect of the FN in all cases of typical HFS except for one. The meatal loop of the anterior inferior cerebellar artery (AICA) compressed the FN when the vessel passed between the FN and the eighth cranial nerve (8th N). These findings suggest that the topographical organization in the FN in the cerebellopontine cistern may be reversed to a peripheral distribution: the fibres on the posterior/rostral side of the FN innervate the lower part of the facial muscles, and those in the anterior/caudal side of the nerve innervate the upper part of the facial muscles. When examining patients with HFS, we must very carefully determine whether patients have typical or atypical HFS, to determine whether blood vessels (usually the meatal loop of the AICA) between the FN and the 8th N as well as at the root exit zone of the FN are to be decompressed. PMID- 9870065 TI - Results of cable graft technique in repair of large nerve trunk lesions. AB - Cable grafting was reintroduced in the beginning of the 1980's as a modified method for repair of large polyfascicular nerves without group arrangement such as trunks and cords of the brachial plexus, sciatic nerve and its divisions, or the other main nerve trunks. We used this method in 25 patients with brachial plexus injury and 29 patients with injuries to the sciatic nerve. Results were analyzed according to the individual nerve elements and were available for 32 elements of the brachial plexus and for 19 tibial and 19 peroneal divisions of the sciatic nerve. We defined useful functional recovery according to the priorities in repair of the brachial plexus and sciatic nerve with lower limits M3 for relevant muscles and functions and S2 for sensory function. Results were especially favourable for the brachial plexus with a total rate of recovery 84% and for tibial division with the same total rate of recovery. On the basis of the results obtained we were able to conclude that the results of the modified cable grafting were at least similar to those of interfascicular nerve grafting and that this method could be useful for repair of large polyfascicular nerve trunks. However, the main advantage of this technique is a considerable gain in operative time. PMID- 9870066 TI - Cisternography in combination with single photon emission tomography for the detection of the leakage site in patients with cerebrospinal fluid rhinorrhea: preliminary report. AB - The site of leakage in a patients with rhinorrhea of various origin may be difficult to identify. The aim of our paper is to evaluate the contribution of cisternography in combination with single photon emission tomography (SPECT) to identify the fistulous track. From 1/1/1992 to 30/11/1997 we studied 20 patients with rhinorrhea posing a challenging diagnostic problem as to identification of the leakage site. Two mls of Indium DTPA (In 111) were injected into the subarachnoid space by the lumbar route. The tracer was followed by planar scintigraphy until it reached the cranial base and subsequently the SPECT acquisition started. A fistula was demonstrated in all of our cases including patients with no active leakage at the time of examination, patients with no bone defects on thin sliced CT scanning or patients with a normal MRI. At surgery the fistulous track was confirmed in all but two cases when a bilateral fistula was operatively identified only on one side. In conclusion whenever a CT scanning fails to demonstrate significant bone defects and MRI does not localize a fistulous track, SPECT cisternography via the lumbar route proved in our experience to be a reliable examination for a precise diagnosis. PMID- 9870067 TI - Primary meningeal malignant fibrous histiocytoma with cerebrospinal dissemination and pulmonary metastasis. AB - Primary intracranial malignant fibrous histiocytoma (MFH) is very rare, and not much is known about its clinical features. The authors report a case of left temporal leptomeningeal MFH, with consequent cerebrospinal fluid (CSF) dissemination and pulmonary metastasis. The clinical features of this case and the therapeutic prognosis of 17 cases reported previously in the literature were reviewed. PMID- 9870068 TI - 5 years' experience with a structured operative training programme for neurosurgical residents. AB - The operative neurosurgical training programme in our clinic was restructured from 1991-93 with the concept of having a frame for the categories and the volume of operations for training year 1 to 6 and to continuously escalate the complexity of the interventions. In the present report the experiences gained so far as well as the deficiencies are described. Between 1991 and 1995 the number of major neurosurgical operations was in the range of 2100 per year, and about 41 48% of these operations were done--under supervision--by residents. By slowly reducing the number of residents from 13 to 9, the trainees started to gain surgical experience earlier, and the average number of operations performed per year increased markedly (from 82 to 122), approximating more to our preplanned figures, also in the various categories. An important aspect is therefore to adapt the number of trainees relative to the available operative case material. According to our preliminary data, about 250-300 operations per year are needed to train adequately one resident. The evaluation also showed deficiencies in some categories, e.g., in pain treatment and peripheral nerve surgery, where care must be taken to better fulfil the official requirements. The object of a 6-year education is to offer a well balanced training programme with systematic escalation of surgical responsibility until full competency is reached. However, this goal needs to be defined more precisely. The plan presented recently by the Committee for Graduate and Postgraduate Education of the German Society of Neurosurgery [1] may serve as a proposal. A personal surgical log-book would allow a much better record and evaluation the progress of the individual trainee as well as the engagement of the teachers. PMID- 9870069 TI - Cranial base perforation caused by a ventriculosubgaleal shunt. PMID- 9870070 TI - Metastatic brain adenocarcinoma masquerading as recurrent haemangioblastoma. PMID- 9870071 TI - Dumb-bell hypoglossal neurinoma with intra- and extracranial paravertebral expansion. PMID- 9870072 TI - Identification of the source of inhibins at the time of conception provides a diagnostic role for them in very early pregnancy. AB - PROBLEM: Early diagnosis of a complicated or poor pregnancy outcome in patients undergoing assisted reproductive technique (ART) fertility treatment could aid their counseling and management. A possible role for the inhibin superfamily as markers of early pregnancy viability was investigated. METHOD OF STUDY: To determine the source of the dimeric glycoproteins inhibin A (alpha-beta A) in early pregnancy, serial blood samples from women who became pregnant following in vitro fertilization (IVF) with fresh embryo transfer (n = 50), from women who achieved pregnancy with frozen-thawed embryos (n = 8), and from a control group of women with spontaneous conceptions (n = 7) were analyzed using a two-site enzyme-linked immunosorbent assay (ELISA). Gonadotropin-releasing hormone (GnRH) analogues are routinely used in ART treatment cycles and are recognized to be luteolytic, and hence, periconceptual administration may be deleterious to pregnancy outcome. Serum samples were obtained from 8 IVF patients who conceived during the cycle in which they had inadvertent luteal phase exposure to GnRH analogues. RESULTS: Elevated serum levels of inhibin A were detected in ongoing pregnancies from 4 weeks' gestation and increased to an initial peak at 9-10 weeks' gestation. Significantly higher levels (P < 0.05) were found in the multiple pregnancies, and nonviable clinical pregnancies had very low levels of inhibin A. Inhibin pro-alpha C was detectable at levels above normal late luteal values in singleton and multiple pregnancies arising from IVF with fresh embryo transfer. In pregnancies established without corpus luteum activity, frozen-thaw embryo replacement, the levels of pro-alpha C containing inhibins were extremely low, suggesting that the corpus luteum is the major source of the alpha monomer. In pregnancies following inadvertent periconceptual exposure to GnRH analogue, the levels of pro-alpha C were statistically significantly higher in successful pregnancies than in early pregnancy failures. CONCLUSIONS: The feto-placental unit is confirmed as the major source of inhibin A in early pregnancy, and the initially low levels and very rapid decline in inhibin A in pregnancies with embryonic failure suggest a role for this glycoprotein as a monitor of early pregnancy viability. The corpus luteum is demonstrated to be the major source of inhibin pro-alpha C in early pregnancy, and very low levels in patients with peri implantational exposure are indicative of lytic damage and herald pregnancy failure despite luteal supplementation with progesterone. PMID- 9870073 TI - Cytokine expression by first-trimester human chorionic villi. AB - PROBLEM: Communication at the human maternal-fetal interface occurs by an intricate cytokine network. This study examines cytokine expression by normal first-trimester human chorionic villi. METHOD OF STUDY: Tissues were obtained at elective pregnancy terminations (7-9 weeks). Total RNA was isolated from chorionic villi by guanidinium isothiocynate-acid phenol extraction. A reverse transcriptase-polymerase chain reaction technique was used to examine cytokine expression. beta-Actin was used as the housekeeping gene, and mitogen-stimulated lymphocytes served as positive controls. RESULTS: beta-Actin was uniformly expressed by all chorionic villous samples. Interferon (IFN)-alpha and -beta also were highly expressed. Moderate expression was noted for interleukin (IL)-10, IL 6, tumor necrosis factor (TNF)-alpha, and IL-1 beta. In contrast, transforming growth factor-beta 1, IFN-gamma, IL-2, and IL-1 alpha were either weakly expressed or absent in first-trimester villi. CONCLUSIONS: Cytokines may contribute to pregnancy immunotolerance (IFN-alpha, IFN-beta, and IL-10), viral resistance (IFNs), hormone secretion (IL-1 and IL-6), and cellular remodeling (IFN-gamma and TNF-alpha) within the chorionic villous. PMID- 9870074 TI - Anti-inflammatory effects of interleukin-4, interleukin-10, and transforming growth factor-beta on human placental cells in vitro. AB - PROBLEM: To determine the effects of anti-inflammatory cytokines on the production of inflammatory mediators by placental cells. METHOD OF STUDY: Cells from term human placentas were isolated and cultured in vitro in the presence of anti-inflammatory cytokines interleukin (IL)-4, IL-10, and transforming growth factor (TGF)-beta. Their effects on the production of IL-8 and prostaglandin E2 (PGE2) were investigated under basal conditions and after stimulation with IL-1 beta and tumor necrosis factor (TNF)-alpha. RESULTS: Both IL-10 and IL-4 inhibited IL-1 beta- and TNF-alpha-induced PGE2 production but had no significant effects on the production of PGE2 under basal conditions. TGF-beta 1 was without effect in stimulated cells, whereas under basal conditions TGF-beta 1 stimulated PGE2 production. Similar trends were seen for IL-8 production, with the exceptions that TGF-beta 1 decreased the TNF-alpha-induced production and IL-4 decreased basal IL-8 production. CONCLUSIONS: The anti-inflammatory effects shown by IL-4, IL-10, and (to lesser extent) TGF-beta may play a role in ameliorating the potentially harmful effects of pro-inflammatory mediators in the feto placental unit. PMID- 9870075 TI - Invasion of cytotrophoblastic (JEG-3) cells is up-regulated by interleukin-15 in vitro. AB - PROBLEM: Trophoblast invasion into the uterus is controlled by many factors. Some cytokines (interleukin [IL]-1, IL-6, and IL-10) have been shown previously to play an important role in placentation. The human placenta is an important source of IL-15, although the cellular source of IL-15 in the placenta has not yet been specified. IL-15 influences cell adhesion and migration by redistributing adhesion molecules in lymphocytes and has been shown to have effects on endothelial cells and in some human tumors. METHOD OF STUDY: To study the role of IL-15 in trophoblast invasion, we investigated the effect of IL-15 (concentrations, 1-10 ng/ml) in a trophoblast invasion model (JEG-3 with matrigel coated filters). Cell invasion was assessed using matrigel-coated filters and was expressed as the quotient of invading cells in comparison with the number of cells that had passed the control membrane. Cell migration was studied by examining the number of cells that had passed the filters without matrigel. Cell proliferation was quantified by a tetrazolium salt WST-1 cleavage assay. Matrix metalloproteinase (MMP)-1, MMP-2, and MMP-9 activities were measured by specific enzyme assays. RESULTS: IL-15 significantly (P < 0.05) increased the in vitro invasion of cytotrophoblastic (JEG-3) cells in a dose-dependent manner. There was a fourfold increase in the invasion at a concentration of 10 ng/ml of IL-15. Migration also was increased by a factor of 2.3 (P < 0.05). Cell proliferation, however, remained unchanged. The collagenolytic activity of cytotrophoblastic (JEG-3) cells was increased by IL-15 stimulation. A significant increase in MMP-1 concentration occurred after the incubation of JEG-3 cells with IL-15. No changes appeared in MMP-2, MMP-9, and tissue inhibitor of metalloproteinase-1 concentrations. CONCLUSIONS: Trophoblast invasion and migration, but not proliferation, are enhanced by IL-15. Our results suggest a role for IL-15 in the modulation of MMP-1 secretion by JEG-3 cells. Furthermore, we speculate, that IL 15 might be related to the changes of cell adhesion molecule phenotype during the process of invasion. PMID- 9870076 TI - Comparative studies on the effects of interleukin-4 and interleukin-13 on cytokine and prostaglandin E2 production by amnion-derived WISH cells. AB - PROBLEM: In hematopoietic cells, interleukin (IL)-13 shares many actions with IL 4. The effects of IL-13 in gestational tissues have yet to be reported, however. We compared the effects of IL-4 and IL-13 on the production of cytokines and prostaglandin E2 (PGE2) in epithelial amnion-derived WISH cells. METHOD OF STUDY: WISH cells were treated with IL-4 or IL-13 (0.08-10 ng/ml) with/without cotreatment with IL-1 beta (0.2 ng/ml), tumor necrosis factor-alpha (10 ng/ml) or epidermal growth factor (5 ng/ml). The production of IL-6, IL-8, and PGE2 was measured by immunoassay after 16 hr. RESULTS: Both IL-4 and IL-13 inhibited PGE2 production with indistinguishable concentration-response curves, under basal or stimulated conditions. The maximal inhibition of IL-1 beta-stimulated PGE2 production (to 28% +/- 10% of control) was seen at 10 ng/ml of IL-4 or IL-13. Basal IL-6 production was stimulated approximately twofold by IL-4 and IL-13, whereas IL-4 and IL-13 both inhibited cytokine-stimulated (but not basal) IL-8 production by approximately 50%. In the presence of 1 ng/ml of IL-4, IL-13 was unable to further inhibit PGE2 production. CONCLUSIONS: The inhibition of PGE2 and IL-8 production by IL-4 in WISH cells is mimicked by IL-13. Both cytokines, probably through binding to a common receptor complex, may share a role in suppressing inflammatory reactions within intrauterine tissues. PMID- 9870077 TI - Gestational age-dependent extravillous cytotrophoblast osteopontin immunolocalization differentiates between normal and preeclamptic pregnancies. AB - PROBLEM: Normal placentation requires modulation of proliferative cytotrophoblast to an invasive phenotype. Preeclampsia is characterized by failed cytotrophoblast invasion and arterial remodeling. Osteopontin (OPN) is an extracellular matrix protein implicated in cell adhesion, spreading, and invasion. METHOD OF STUDY: To investigate gestational age-related OPN expression, placental immunostaining was performed. To investigate the role of OPN in uteroplacental vascular pathology, placental immunostaining from pregnancies with preeclampsia (n = 12), fetal growth retardation (FGR) (n = 8), or both (n = 4) was compared with gestational age-matched controls (n = 24). RESULTS: In non-preeclamptic pregnancies, OPN immunolocalized to basal plate and intervillous cytotrophoblasts from 24-30 weeks (n = 13). In preeclampsia, OPN immunoreactivity was detected from 24-40 weeks. Cytotrophoblasts from FGR placentas were OPN-positive until 30 weeks, unless preeclampsia accompanied the FGR. In this case, cytotrophoblasts were OPN positive from 24-40 weeks. CONCLUSIONS: The data suggest a role for OPN in cytotrophoblast invasion of the maternal vasculature/extracellular matrix during non-preeclamptic placentation, and OPN may serve as a marker for placental bed remodeling. PMID- 9870078 TI - Transport of proteins across the human placenta. AB - PROBLEM: The transport of various proteins across the human placenta was investigated by comparing maternal and fetal concentrations of tetanus antigen (TT-AG), anti-tetanus (TT)-immunoglobulin G (IgG) (following maternal vaccination), IgA, human chorionic gonadotropin (hCG), human placental lactogen (hPL), and alpha-fetoprotein (AFP) at term. METHOD OF STUDY: The concentrations of the six proteins were determined using enzyme-linked immunosorbent assay in serum of maternal venous and umbilical (fetal) vein samples obtained at delivery from uncomplicated term pregnancies (n = 16). RESULTS: The ratios (mean +/- standard deviation) of fetal (umbilical) to maternal level were 1.41 +/- 0.33 (anti-TT-IgG), 0.91 +/- 0.37 (TT-AG), 0.002 +/- 0.001 (IgA), 0.003 +/- 0.001 (hCG), and 0.008 +/- 0.004 (hPL), while the maternal:fetal concentration ratio of AFP was 0.002 +/- 0.002. IgA, hCG, hPL, and AFP showed a close correlation between maternal and fetal levels varying between r2 = 0.47 to 0.73 (P < 0.004 0.0001). Because AFP is produced by the fetus while IgA originates in the mother, the appearance of small amounts of these two proteins in the maternal or fetal compartment, respectively, suggests a slow rate of diffusion following a high concentration gradient. The detection of hCG and hPL in fetal serum is also interpreted as diffusion from the maternal into the fetal blood. Anti-TT-IgG has a significantly higher concentration in the fetal as compared with the maternal serum, which is in line with the well-documented active transfer of IgG. Fetal TT antigen levels were similar to maternal concentrations, showing a close correlation (r2 = 0.74, P < 0.0001) between the two proteins. CONCLUSIONS: The correlation between maternal and fetal concentrations of various proteins like IgA (150,000 Da), hCG (42,000 Da), and hPL (21,000 Da) suggests passive diffusion of these macromolecules across the placenta from the maternal to the fetal side, albeit at a slow rate. A similar process is postulated for AFP (70,000 Da) diffusing in the opposite direction from the fetus to the mother. There was no significant difference between the transplacental fetomaternal gradient of IgA and hCG and the maternal-fetal gradient of AFP. In view of the substantially larger volume of circulating maternal as compared with fetal blood, a significantly higher rate of crossing of AFP as compared with the other proteins must be assumed. It is uncertain whether a difference in the rate of transplacental transfer in the two directions or an additional source of AFP production in the maternal compartment explains the high maternal level. Anti-TT IgG concentration is significantly higher in fetal than in maternal serum suggesting active transfer from the mother to the fetus. Furthermore, there is considerable transfer of TT-AG and a close correlation of fetal:maternal ratios of anti-TT-IgG (150,000 Da) and TT-AG (150,000 Da) could be an indication for a specific transfer of the antigen antibody complex. PMID- 9870079 TI - Natural killer cell cytotoxicity and paternal lymphocyte immunization in women with recurrent spontaneous abortions. AB - PROBLEM: Natural killer (NK)-cell cytotoxicity in women undergoing lymphocyte immunization prior to and following treatment was investigated. METHOD OF STUDY: A cohort of 33 women with a history of two or more recurrent spontaneous abortions was prospectively studied. NK-cell cytotoxicity was determined at effector-to-target ratios of 50:1 and 25:1. Peripheral blood CD56+ NK-cell, CD19+ B-cell, CD19+/5+ B-1-cell, and CD3+ pan T-cell levels were studied by flow cytometry before and after lymphocyte immunization treatment. Maternal antipaternal T- and B-cell antibody levels were measured before and after lymphocyte immunization by flow cytometric analysis. Paternal lymphocyte immunizations were given two times with a 4-week interval. Post-lymphocyte immunization testing was done 4 weeks after the second lymphocyte immunization. The controls were 8 normal healthy women. NK assays were done twice with an interval of 8 weeks. RESULTS: NK-cell activity at effector-to-target ratios of 50:1 (P = 0.005) and 25:1 (P = 0.001) were significantly suppressed after lymphocyte immunization. CD3+ pan T-cell levels after lymphocyte immunization were significantly increased compared with levels before lymphocyte immunization (P = 0.008). CD56+ NK-cell levels were significantly suppressed after lymphocyte immunization (P = 0.016). There was no correlation between changes in NK cytotoxicity and differences in antipaternal lymphocyte antibody levels before or after lymphocyte immunization. CONCLUSION: Lymphocyte immunization suppresses NK cell cytotoxicity and CD56+ NK-cell levels and increases the peripheral blood CD3+ T-cell population in women with recurrent spontaneous abortions. PMID- 9870080 TI - Gonadotropins do not induce antiphospholipid antibodies. AB - PROBLEM: To determine whether the increased incidence of antiphospholipid antibodies (APAs) in women undergoing assisted reproduction might be secondary to superovulation with gonadotropins, predisposing women to an abnormal immune response and thus inducing APAs. METHOD OF STUDY: Women undergoing assisted reproduction with gonadotropins for the first time were selected and tested before the initiation of the stimulation cycle, during the cycle, and at the end of the cycle (group 1). Women who had undergone gonadotropin stimulation at least 60 days earlier (group 2) and normal, nonpregnant, fertile women (group 3) also were evaluated. Serum samples were assayed by the enzyme-linked immunosorbent assay method. RESULTS: Ten (20%) of 50 women in group 1 were positive for APAs. The 10 women who were positive for APAs remained positive throughout the treatment cycle. Positive antibodies were identified in 12 (24%) of 50 women in group 2, not significantly different from group 1 (P = 0.81). Antibodies were present in 2 of 50 normal fertile control subjects, significantly less frequently than in group 1 (P < 0.03) and in group 2 (P < 0.01). CONCLUSIONS: These data suggest that gonadotropin administration and/or the ovarian response to stimulation does not predispose women to the induction of APAs. Moreover, the incidence of APAs in this population, which is higher than that found in normal fertile women, cannot be explained by cycle-induced events. PMID- 9870081 TI - Novel implications in the development of endometriosis: biphasic effect of macrophage activation on peritoneal tissue expression of tissue inhibitor of metalloproteinase-1. AB - PROBLEM: Elevated levels of activated macrophages are associated with endometriosis, but their role in the etiology of the disease is uncertain. The current study was undertaken to examine whether activated macrophages could modulate peritoneal tissue expression of the tissue inhibitor of metalloproteinase-1 (TIMP-1), which may play a role in the development of endometriosis. METHOD OF STUDY: Female mice were treated with the macrophage activator lipopolysaccharide (LPS), and peritoneum TIMP-1 mRNA was examined by Northern blot analysis. RESULTS: LPS induced a dose-dependent increase (P < 0.05) in TIMP-1 mRNA expression at levels of 1 microgram (70.0% +/- 5.8% greater than the control), 10 micrograms (83.0% +/- 12.0% greater than the control), and 25 micrograms (100.0% +/- 10.0% greater than the control). In contrast, the administration of 50 micrograms of LPS resulted in a decrease in TIMP-1 mRNA expression below baseline levels (18.0% +/- 6.0% less than the control values). CONCLUSIONS: Activated macrophages and/or their products modulate peritoneum TIMP 1 expression. These data suggest that, in addition to their phagocytotic role in the peritoneal cavity, these immune cells also may play a novel role in influencing the ability of the peritoneum to regulate tissue/cell invasion and in the development of endometriosis through TIMP-1 expression. PMID- 9870083 TI - Granulocyte macrophage-colony stimulating factor production by autologous endometrial co-culture is associated with outcome for in vitro fertilization patients with a history of multiple implantation failures. AB - PROBLEM: To determine whether granulocyte macrophage (GM)-colony stimulating factor (CSF) produced by autologous endometrial co-culture was associated with outcome in 53 patients with a history of multiple in vitro fertilization failures. METHOD OF STUDY: The conditioned media from endometrial co-culture cells exposed or non-exposed to human embryos was analyzed for GM-CSF. RESULTS: Exposure or non-exposure to an embryo did not result in an enhancement of GM-CSF production. Insignificant levels of GM-CSF were determined from media alone. ROC analysis revealed that levels of GM-CSF from supernatants of endometrial co culture exposed to embryos that measured below 130 pg/ml reflected a diminished prognosis (5/17 had a positive pregnancy vs. 21/36 with GM-CSF levels greater than 130 pg/ml; P < 0.05). CONCLUSIONS: The improved outcome associated with GM CSF values greater than 130 pg/ml may reflect: 1) a direct positive effect of GM CSF; 2) an embryotrophic factor upregulated by GM-CSF; or, 3) that GM-CSF functions as a marker for the importance of the glandular component in endometrial co-culture systems. PMID- 9870082 TI - Endothelin-1: expression and role in human corpus luteum. AB - PROBLEM: Several recent data suggest an involvement of endothelin (ET)-1, a powerful vasoconstrictor peptide, in reproductive function. This study was designed to investigate the presence and role of ET-1 in human corpus luteum. METHOD OF STUDY: Purified luteal cells were incubated for different times with ET 1 or ET-3 alone or associated with human chorionic gonadotropin. In another set of experiments cells were treated with ET-1 and BQ485, an ET-A receptor antagonist, or with phorbol 12-myristate-13 acetate (PMA), an activator of protein kinase C. RESULTS: ET-1 reduced both basal and human chorionic gonadotropin-induced progesterone production at all examined times, similarly PMA inhibited basal progesterone synthesis. BQ485 prevented the inhibitory effect of ET-1, while no effect was observed with ET-3. Finally, ET-1 mRNA was detected in the luteal cells. CONCLUSION: ET-1 is expressed by human luteal cells and reduces basal and human chorionic gonadotropin-induced progesterone synthesis through the ET-A receptors and the protein kinase C pathway. Conversely, ET-3 does not affect luteal steroidogenesis. PMID- 9870084 TI - [Echocardiography. Its role in preoperative diagnosis of cardiac risk patients before non-cardiac surgical interventions]. PMID- 9870085 TI - [The role of echocardiography in preoperative diagnosis of cardiac risk in patients before non-cardiac surgical interventions]. AB - Echocardiography is a noninvasive method for cardiac evaluation. A review of the current literature shows that the routine use of echocardiography for assessing perioperative cardiac risk in patients undergoing noncardiac surgery can not be supported. Only patients with suspected relevant heart valve diseases, acute heart failure, cardiomyopathy or condition after heart or heart-lung transplantation may benefit from preoperative echocardiography. In patients with suspected or proven coronary artery disease stress echocardiography offers the most relevant additional information for the anaesthesiologist. However, because of the high financial and personal implications it should be reserved to those patients who are not able to perform a normal stress test. Besides in patients in whom transthoracic echocardiography doesn't offer sufficient information or is not possible transesophageal echocardiography plays only a minor role in preoperative cardiac evaluation. PMID- 9870086 TI - [Retrospective analysis of transpulmonary and pulmonary arterial measurement of cardiac output in ARDS patients]. AB - OBJECTIVES: To investigate the agreement (and its potential dependency on extra vascular lung water) between transpulmonary (TPID) and standard pulmonary artery (PAID) thermodilution cardiac output measurements. METHODS: One hundred and sixty simultaneous cardiac output measurements using transpulmonary and pulmonary artery thermodilution techniques were retrospectively compared in 18 patients with acute respiratory distress syndrome. In addition, extravascular lung water was determined using a double indicator technique (temperature and indocyanine green). RESULTS: Mean (+/- SD) difference ("bias") was 0.03 L/min (+/- 1.04 L/min), linear regression analysis resulted in TPID = 0.87 PAID + 1.16 (r = 0.91). Mean extra vascular lung water was 1625 mL (minimum-maximum: 403-3266 mL) and therefore markedly elevated as could have been expected in patients with ARDS. Bias (PAID-TPID) was not dependent on extravascular lung water. CONCLUSIONS: Transpulmonary and pulmonary artery thermodilution methods can be used interchangeably. The results demonstrate for the first time in humans that transpulmonary thermodilution provides valid cardiac output values in patients with markedly increased fluid content of the lungs. PMID- 9870087 TI - [Antiemetic effect of propofol]. AB - Propofol is known to possess direct antiemetic effects. Its use for induction and maintenance of anaesthesia has been shown to be associated with a lower incidence of postoperative nausea and vomiting (PONV) when compared to any other anaesthetic drug or technique. However, its mechanism of action in this context is still not well understood. In this article, the best ways to take advantage of propofol's antiemetic properties are emphasized. The possible effects of propofol on the cerebral cortex, its interactions with the dopaminergic and the serotoninergic systems are evaluated by the known clinical and basic science results. Finally, the advantages and disadvantages of using propofol to decrease the incidence of PONV in clinical practice are discussed. PMID- 9870088 TI - [The official American guidelines for prevention of nosocomial pneumonia. U. S. Centers for Disease Control]. AB - The official German guidelines for prevention of nosocomial pneumonia were published by the Bundesgesundheitsamt, now called Robert-Koch-Institut, twelve years ago. The recently published official "guidelines for prevention of nosocomial pneumonia" of the Centers for Disease Control and Prevention (CDC) are categorized according to scientific evidence. The American guidelines are very detailed and differ in some aspects from the official German guidelines. The purpose of the present paper is to inform the German anaesthesiologist about the official CDC guidelines and to provide a renewed background for the prevention of nosocomial pneumonia. PMID- 9870089 TI - [Reduced neuromuscular blocking potency of atracurium in patients with purulent intrathoracic diseases]. AB - OBJECTIVE: Based on personal observations the neuromuscular blocking potency of atracurium was supposed to be diminished in purulent intrathoracic diseases. This hypothesis was tested in a prospective clinical trial. METHODS: 52 adult patients undergoing general anaesthesia (methohexitone, sufentanil, flunitrazepam, N2O, enflurane) for elective thoracic surgery were investigated. After the intubation dose of 0.6 mg/kg atracurium was applied continuously to maintain a 90% suppression of the evoked compound electromyogram. According to the intraoperatively established diagnosis patients were allocated to three categories: 1) non-malignant tumor as the control group (n = 15), 2) lung cancer (n = 22), 3) purulent intrathoracic process without tumor (n = 15). The groups were compared regarding onset time, DUR 10% and maintenance dose of atracurium. RESULTS: Patients with lung cancer did not differ significantly from the controls regarding efficiency of atracurium. In contrast, patients with a purulent intrathoracic process showed a significantly longer onset time (6.3 +/- 2.5 vs. 2.9 +/- 0.8 min, p < 0.001), and a significantly shorter DUR 10% (23 +/- 6 vs. 36 +/- 10 min, p < 0.001) compared to the control group. Mean infusion rate of atracurium to maintain a 90% suppression of the evoked compound electromyogram was significantly higher in patients with a purulent process compared to the controls (10.5 +/- 3.2 vs. 6.0 +/- 1.2 micrograms/kg.min, p < 0.001). CONCLUSION: Our results support the hypothesis that patients with a purulent intrathoracic disease show a clear reduction in neuromuscular blocking potency of atracurium. PMID- 9870090 TI - [Anesthesia and perioperative immune function. Remarks on the work of M. Bauer, H. Rensing and T. Ziegenfuss (Anaesthesist 1998,47:538-556)]. PMID- 9870091 TI - [Interscalene block and infections of the shoulder]. PMID- 9870093 TI - [Adjuvant therapy for sepsis and ARDS]. PMID- 9870092 TI - [Peridural administration of lipophilic opioids]. PMID- 9870094 TI - [Documentation of opiate consumption]. PMID- 9870096 TI - [What is the "ideal colloid"? New aspects of volume therapy]. PMID- 9870095 TI - [The drug emergency]. PMID- 9870098 TI - Laser scanning cytometry quantification of estrogen receptors in breast cancer. AB - OBJECTIVE: To describe the laser scanning cytometry (LSC) processing and analysis developed for the quantitative analysis of estrogen receptor (ER) content in routine paraffin sections of breast carcinomas. STUDY DESIGN: Histologic sections of archival, paraffin-embedded tissues from 30 breast carcinomas were labeled for ER with fluoresceinated monoclonal antibody. ER expression was quantified by LSC and expressed as percent positive tumor cells and as histogram distributions of receptor expression per cell. Duplicate sections of the same tumors were stained for ER by a conventional immunoperoxidase reaction and percent positive tumor cells counted visually. RESULTS: Percent ER-positive tumor cells by LSC of immunofluorescence-stained sections correlated well with conventional (visual) counts of immunoperoxidase-stained duplicate sections when the latter were categorized as low, intermediate or high percent of positive cells. In addition, the marked variation in relative number of ER binding sites per cell could be quantified by LSC and displayed in histogram distribution. CONCLUSION: LSC measurements are fast and objective and can be carried out on sections of paraffin-embedded tissue after routine processing in the pathology laboratory. In addition, LSC data provide the relative number of ER binding sites per unit of DNA; that may reveal clinically significant skewed distributions or subpopulations of tumor cells. PMID- 9870097 TI - Transforming growth factor-beta expression in human testicular neoplasms. AB - OBJECTIVE: To investigate the localization of transforming growth factors (TGF beta 1, -beta 2 and -beta 3) and their receptors (TGF-beta RI and RII). STUDY DESIGN: The study included 26 paraffin-embedded tissues from human testicular neoplasms: 15 seminomas, 2 embryonal carcinomas, 1 immature teratoma, 4 immature teratomas with embryonal carcinoma, 1 immature teratoma with seminoma, 1 seminoma with embryonal carcinoma and 2 gonadal stromal tumors (Leydig cell tumors). RESULTS: TGF-beta 1 immunoreactivity was cytoplasmic and was expressed in 22 (84.6%), TGF-beta 2 in 20 (77%), TGF-beta 3 in 11 (42.3%), TGF-beta-RI in 21 (80.8%) and TGF-beta-RII in 18 (69.2%) of the 26 neoplasms. The percentage of positive immunostained cells and the intensity of staining were significantly higher in tumor than in peritumor nonneoplastic testis. In the peritumor nonneoplastic testis, Leydig, Sertoli and germ cells coexpressed both the three TGF-beta isoforms and TGF-beta-RI and RII. The myoepithelial cells of the seminiferous tubules showed immunoreactivity for TGF-beta RI and RII but not for TGF-beta s. In tumor testis areas the pattern of TGF-beta and TGF-beta receptor expression and distribution varied according to the histologic type of testicular tumor. Seminomas showed a diffuse pattern of TGF-beta immunoreactivity, whereas immature teratomas had focal and patchy distribution. In teratomas, differentiated structures contained more TGF-beta s than undifferentiated structures. PMID- 9870099 TI - Characterization of cosmids and telomeres in cytogenetic preparations by 3D confocal fluorescence. AB - OBJECTIVE: To analyze, with fluorescent probes, by three-dimensional (3D) emission patterns, fluorescence in situ hybridization (FISH) preparations (cosmids, telomeres) and to perform factor analysis of medical image sequences (FAMIS); to use FISH to track relevant DNA sequences in cell nuclei during interphase and in mitotic chromosomes; and to use cytogenetic techniques, resulting in flat preparations of whole cells that are assumed to preserve probe access to their targets. STUDY DESIGN: The study design entailed labeling targets by probes (sequences labeled by fluorescein isothiocyanate) in nuclei and/or chromosomes stained by propidium iodide. Visualization of targets was improved when 3D sequences of images obtained on a single photomultiplier detector of the confocal microscope by z stepping were investigated by FAMIS. RESULTS: Factors and factor images showed that targets could be detected and differentiated in focal planes inside nuclei or chromosomes. CONCLUSION: It is possible to localize cosmids in cell nuclei at interphase and telomeres in mitotic chromosomes by means of 3D sequences of images. PMID- 9870100 TI - Morphometric analysis of AgNORs in uveal malignant melanoma. AB - OBJECTIVE: To define the value of silver-stained nucleolar organizer regions (AgNOR) in the evaluation of uveal malignant melanomas (MM). STUDY DESIGN: Morphometric analysis of AgNORs was performed on 30 uveal MM of spindle A, B, epithelioid and mixed cell types. Findings were compared with cytotype, largest tumor dimension (LTD) and clinical outcome. RESULTS: AgNOR mean perimeter (MP) was 3.76 microns (SD 1.21, range 1.79-8.51 microns) in A, 5.05 microns (SD 1.96, range 1.79-16.41 microns) in B and 6.15 microns (SD 3.86, range 1.79-33.80 microns) in epithelioid MM; mean area (MA) was 0.86 micron 2 (SD 0.60, range 0.20 2.78 microns 2) in A, 1.58 microns 2 (SD 1.11, range 0.20-9.63 microns 2) in B and 2.34 microns 2 (SD 2.10, range 0.20-15.27 microns 2) in epithelioid MM; the aspect ratio (AR) was 3.10 (SD 0.19, range 2.90-3.35) in A, 5.80 (SD 2.07, range 3.77-9.32) in B and 12.22 (SD 2.84, range 8.11-15.81) in epithelioid MM. From comparing MA and RA with follow-up, it seemed that MM with a good prognosis (spindle A and a subgroup of B tumors, B1) exhibited the lowest MA and AR values, while MM with poorer clinical behavior (epithelioid and a second subgroup of spindle B MM, B2) showed the highest. No correlation was found between AgNORs and LTD. CONCLUSION: AgNORs may contribute to the prognostic evaluation of uveal MM. PMID- 9870101 TI - Image analysis of dermal collagen changes during skin aging. AB - OBJECTIVE: To determine progressive quantitative, directional and textural changes in dermal collagen as a function of age and sex and to estimate their evolutive trend with appropriate regression models. STUDY DESIGN: Ninety-six samples of abdominal skin from autopsy cases were analyzed. The ages ranged from 3.5 months to 86 years. Picro-Sirius-stained slides were examined by polarizing microscopy, and spatial density, directional features and texture of collagen were measured by computerized image analysis. Nonlinear regression models were built to estimate evolutive changes with respect to age. The relationship between spatial orientation of collagen bundles and age was best modeled by linear regression. RESULTS: The evolutive patterns of dermal thickness and spatial density of collagen bundles correspond to a second-order polynomial model with a progressive increase from childhood to middle age and a relatively sharp decrease after the seventh decade. The evolution of textural pattern of dermal collagen, defined by gradient analysis, depicts a sort of inverted U. Its complexity is maximum in the first year of life, decreases until the period 25-50 years and increases progressively after the sixth decade. The horizontal orientation of collagen bundles with intermingled fascicles oriented in other directions, shown by young individuals, is progressively simplified with aging. In elderly subjects, collagen bundles have a horizontal orientation. No significant sex related differences were found. CONCLUSION: Dermal collagen changes related to aging are apparently independent of sex, at least in abdominal skin, and show characteristic curvilinear evolutive trends defined by decreased dermal thickness in the elderly, decrease in the spatial density of collagen bundles and increase in textural heterogeneity of the dermis. Progressive simplification in the orientation of collagen bundles leading to a predominant horizontal disposition followed a linear trend. These changes could contribute to providing a substantial morphologic basis to age-associated biomechanical alterations in the skin. PMID- 9870102 TI - Discrimination of esophageal dysplasia with progression and nonprogression. High resolution image analysis for surrogate end point biomarkers. AB - OBJECTIVE: To detect textural nuclear features correlated with nonprogression and progression in esophageal dysplasia. STUDY DESIGN: Asymptomatic adults from Heshun Commune, Linxian County, China were examined with a balloon sampler in 1983 fifty cases of moderate esophageal dysplasia and 68 cases of mild were selected for study. By means of an Axiomat microscope equipped with a TV camera, 100 visually normal intermediate squamous cell nuclei per specimen were randomly measured from routinely Papanicolaou-stained slides. RESULTS: Of 50 esophageal moderate dysplasia cases, 24 and 7 progressed to carcinoma within three and nine years, respectively. The other 19 cases remained stable or regressed to normal and were used as the control group. By means of chromatin features, correct specimen classification rates of 79.2% (19/24), 73.7% (14/19), 85.5% (6/7) and 84.2% (16/19) were achieved, respectively (P < .001). Of 68 cases classified as mild dysplasia, 16, 13 and 12 progressed to carcinoma within three, five and nine years, respectively. The other 27 cases remained stable or regressed to normal and were used as the control group. The correct specimen classification rates were 93.8% (15/16), 88.9% (24/27), 69.2% (9/13), 74.1% (20/27), 83.3% (10/12) and 77.8% (21/27), respectively, using chromatin features of the nuclei (P < .001). CONCLUSION: In this study, nuclear chromatin features measured by high-resolution image analysis could sufficiently well forecast the outcome of precancerous lesions and discriminate precancerous lesions with progression and nonprogression. It also can be employed as surrogate end point biomarkers in clinical chemoprevention trials. Stoichiometric staining and standard preparations should increase the correct classification rates in further studies. PMID- 9870103 TI - Use of image cytometry to classify biliary and ampullary adenocarcinomas. AB - OBJECTIVE: To create an objective classification system to perform TNM classification of ampullary adenocarcinoma and cholangiocarcinoma using image cytometric data derived from Feulgen-stained tumor nuclei. STUDY DESIGN: Surgically resected cases of ampullary adenocarcinoma and cholangiocarcinoma with established TNM classifications were selected on the basis of available formalin fixed, paraffin-embedded tissue. Fifteen numerical variables related to morphometric, densitometric and textural features of each tumor nucleus were recorded. We employed a methodology based on multivariate statistical tools to characterize the association of morphonuclear variables with TNM classification. The first step consisted of identifying and selecting representative nuclei of each T class. From this "purified" data set an objective classification system was created. The classification system was assessed using internal and external validation. RESULTS: Employing ANOVA, all 15 variables were significantly associated with T classification, 11 of 15 with N and 4 with M. Multivariate analysis was employed to distinguish between T1, T2 and T3 lesions. Our methodology correctly classified 76% of T1 nuclei, 47% of T2 nuclei and 84% of T3 nuclei. Heterogeneity within an individual tumor was defined in 61% of cases included in the training set. Complete concordance between pathologic classification and the classification system was observed in 71% of an independent validation. PMID- 9870104 TI - Effect of age on synaptic size in human brain tissue proximal to tumor masses. AB - OBJECTIVE: To measure the effect of age on synaptic morphologic rearrangements in human brain tissue peripheral to space-occupying lesions. STUDY DESIGN: Synaptic length (L) was measured interactively by computer-assisted image analysis in brain tissue samples from adult (mean age, 39 years) and old (mean age, 67.2 years) patients. Each group consisted of five subjects. RESULTS: L was reduced by 9% in the old vs. adult group of patients. A percent distribution of the data showed that junctional areas smaller than 0.25 micron accounted for 47.2% in old patients and 31% in the adults. CONCLUSION: The present findings are in contrast with current literature data documenting a significant increase in the percent of enlarged contact zones in the senile brain. We interpret these results in terms of synaptic dynamic morphology and suggest that they may reflect alterations of neuronal adaptive capabilities due both to the pathologic condition and age of the patients. PMID- 9870105 TI - Quantitative cytochemistry of succinic dehydrogenase activity in rat mitochondria. AB - OBJECTIVE: To set up a computer-assisted morphometric procedure to cytochemically measure the activity of succinic dehydrogenase (SDH). STUDY DESIGN: In rat mitochondria from Purkinje cell perikarya and frozen muscle samples, SDH activity was selectively evidenced by the copper ferrocyanide method. On the SDH-positive organelles we measured the following parameters: number of mitochondria per cubic micrometer (numeric density [Nv]), volume fraction of organelles per cubic micrometer (volume density [Vv]), average mitochondrial volume (V) and intramitochondrial area density of the SDH reaction (area of the precipitates/total mitochondrial area [R]). RESULTS: In both fresh Purkinje cells and frozen muscle cells the positive organelles were sharply evident. By considering Vv, Nv and V altogether in a given experimental group, a reliable evaluation of the morphologic rearrangements of the cellular metabolic hardware can be obtained. Measurements of R provides information on the functional efficiency of each organelle. CONCLUSION: Quantitation of SDH-positive mitochondria is closely associated with the amount of enzyme molecules present and active within the organelles. Thus, quantitative assessment of the copper ferrocyanide reaction contributes to the evaluation of mitochondrial metabolic competence. PMID- 9870106 TI - 2-D index as a measure for estimating the complexity degree of 3-D sinusoidal structures in the normal and cirrhotic liver and in hepatocellular carcinoma. AB - OBJECTIVE: To evaluate the usefulness of a two-dimensional (2-D) index from a single tissue section as a measure for estimating the complexity degree of three dimensional (3-D) sinusoidal structure in normal and cirrhotic liver and in hepatocellular carcinoma (HCC). STUDY DESIGN: The number of sinusoidal profiles, or 2-D index, in a given square area 400 x 400 microns 2 on a single tissue section was counted. The number was compared with the first Betti number of 3-D sinusoidal structures, obtained from previous reconstruction studies. RESULTS: The mean number of sinusoidal profiles (+/- SD) was 104.2 +/- 13.1 in 5 normal livers, 77.0 +/- 15.8 in 10 cirrhotic livers and 34.0 +/- 10.7 in 10 HCCs. Each difference between the three groups was statistically significant (P < .01). The number of sinusoidal profiles was approximately linearly related to the first Betti number of the sinusoids. The correlation coefficient = .77. CONCLUSION: The 2-D index, or number of sinusoidal profiles in a given area on a single tissue section, was useful for estimating the degree of complexity of the 3-D sinusoidal network. PMID- 9870107 TI - Confocal microscopy. Applications in research and practice of pathology. AB - OBJECTIVE: To discuss recent advances in the use of confocal microscopy to perform quantitative analysis of physiologic and pathologic intracellular events in the living organ. STUDY DESIGN: We developed a new confocal microscope system equipped with a microlens-arrayed multipinhole scanning disc and water-immersion objective lenses. The confocal microscope enables us to quantitate nonhomogeneous changes in the three-dimensional intracellular calcium ion concentration in a living, whole heart. RESULTS: The new system allows in situ quantitative imaging of the calcium wave in the living heart at a full video rate. Calcium waves propagating from cell to cell were frequently interrupted by calcium transients from spontaneous sinus rhythm, but low transparency of the living tissues and cell injury with laser light still remained unsolved. CONCLUSION: The evolution of the confocal microscope with high spatial and temporal resolution renders the investigation of biologic phenomena even in the living organ. PMID- 9870109 TI - Two solitudes of complementary and conventional medicine. More on roles for family physicians. PMID- 9870108 TI - Current issues in cardiac care. PMID- 9870110 TI - Hazing at the college. PMID- 9870111 TI - Proper technique for Pap smears. PMID- 9870112 TI - [Vas deferens occlusion: preference or science?]. PMID- 9870113 TI - Do silicone breast implants affect breastfeeding? AB - QUESTION: One of my patients has silicone breast implants. She gave birth to a healthy baby boy but is afraid to breastfeed because she read in a magazine that the baby might be affected by silicone secreted in the milk. How should I advise her? ANSWER: Neither silicone nor its components could be measured in the breast milk of women with silicone implants. Your patient should be able to breastfeed safely. PMID- 9870114 TI - Ophthaproblem. Valsalva retinopathy. PMID- 9870115 TI - Dermacase. Ichthyosis vulgaris. PMID- 9870116 TI - Medicolegal file. Spousal abuse. PMID- 9870117 TI - Practice tips. Medication flow sheet. Useful tool for keeping track of patients' prescriptions. PMID- 9870119 TI - Are the new selective alpha-blockers better than non-selective alpha-blockers for benign prostatic hyperplasia? PMID- 9870118 TI - Do intensive blood pressure lowering and low-dose ASA help our hypertensive patients? PMID- 9870120 TI - Where do family practice patients go in case of emergency? AB - OBJECTIVE: To determine the annual incidence of patient-defined emergencies and patients' use of emergency services at a family medicine teaching unit. DESIGN: Cross-sectional survey. SETTING: Hospital-based family medicine teaching unit in Montreal. PARTICIPANTS: Registered patients attending a family medicine teaching unit during 11 consecutive weekdays. MAIN OUTCOME MEASURES: Annual incidence of patient-defined medical emergencies and use of emergency services at the unit. RESULTS: Eligible patients made 815 visits during the study period; 584 usable questionnaires were returned for a response rate of 71.7%. In the previous 12 months, 37% of patients reported at least one medical emergency. For their last emergency, 42% reported using at least one of the emergency services offered by the clinic. Only 19% of patients with after-hours emergencies reported using our on-call system. Although socioeconomic and clinical variables did not predict the incidence of patient-defined emergencies, multivariate analysis revealed three significant predictors for use: patients with the practice for 5 or more years were more likely to use our services, while patients 75 and older and those with emergencies after hours were less likely to use our services. CONCLUSIONS: In an urban group family practice, annual incidence of medical emergencies among registered patients was 37%. Those whose most recent emergency occurred after hours used the clinics' emergency on-call services disappointingly little. PMID- 9870121 TI - Prospective care of elderly patients in family practice. Is screening effective? AB - OBJECTIVE: To evaluate cost and benefits of screening for and treating health and lifestyle risks among community-dwelling elderly. DESIGN: Randomized controlled trial. SETTING: Primary care. PARTICIPANTS: An opportunistic and prompted sample of 619 rostered elderly patients presenting for treatment who screened positive. INTERVENTIONS: One third (209) of experimental subjects had screening questionnaires placed in their charts with concerns highlighted for referrals. Two control groups received usual care. MAIN OUTCOME MEASURES: Yearly assessments of health service use and multidimensional functional capacity. RESULTS: Overall, screening and treatment of functionally active, elderly, middle-class people had no significant beneficial effect. Almost half of the experimental sample was ineligible because of treatment noncompliance. Generally ineligible subjects were older and more severely impaired. Subjects 75 years and older with risk factors showed improvement in daily living activities, and those living alone were found to have improved mental health and social functions (11% and 22%, respectively). CONCLUSIONS: Screening and treatment was ineffective in improving total functional capacity of all seniors 65 years and older. Elderly people 75 years and older, however, who were living alone or lonely did benefit from screening and treatment showing an improvement in daily activities, mental health scores, and social functions. This finding has implications for selective preventive health care spending for the elderly. A 2-year follow-up period could be too brief to detect long-term effects of early intervention with younger, middle class seniors, especially those who are already functionally active. PMID- 9870122 TI - Community management of heart failure. AB - OBJECTIVE: To review therapies for treating patients with heart failure (HF). QUALITY OF EVIDENCE: Recommendations in this paper are mainly based on the results of randomized controlled trials. To a lesser extent, data from smaller, more physiologic studies are included. Where appropriate, recommendations are based on the results of a consensus conference. MAIN MESSAGE: Although pharmacologic therapy is the main strategy for treating HF patients, general measures, such as counseling and advice about regular physical activity, are an important component of management. Use of angiotensin-converting enzyme inhibitors (ACE-I) is central to treating HF patients, because these agents decrease mortality and morbidity significantly. Digoxin does not reduce mortality but does reduce morbidity. Angiotensin II antagonists, although found to provide clinical benefit equal to ACE-I, have not been found as yet to have similar effects on mortality and morbidity. Diuretics and nitrates are useful for treating these patients' symptoms. Calcium channel blockers should generally be avoided. CONCLUSIONS: Angiotensin-converting enzyme inhibitors are the therapy of choice for HF patients and should be used in all cases unless there are contraindications or clear evidence of intolerance. All other therapies are used mainly for symptom relief. PMID- 9870123 TI - Echocardiography for primary care evaluation of hypertension. AB - OBJECTIVE: To reexamine the changing role of echocardiography for primary care evaluation of essential hypertension since the 1993 Canadian Hypertension Society Consensus Conference. QUALITY OF EVIDENCE: New recommendations from the Canadian Cardiovascular Society and the Canadian Hypertension Society. Recommendations from the American College of Cardiology based on a prospective, randomized controlled trial of limited echocardiograms. Further results from the Framingham Heart Study and the Treatment of Mild Hypertension Study. MAIN MESSAGE: For patients with essential hypertension, left ventricular hypertrophy (LVH) continues to be a strong predictor of many cardiovascular end points, and echocardiography is a much more sensitive tool for detecting LVH than electrocardiogram (ECG). Although limited echocardiograms are not usually used, they are a less costly method of evaluating LVH than standard echocardiograms and have recently been recommended in the United States to assist in the decision to initiate pharmacotherapy for patients with stage 1 hypertension and no other evidence of target organ damage. Problems with intrareader and interreader variability of results are being addressed by technical and mathematical standards established by the Canadian Cardiovascular Society and the Canadian Hypertension Society. CONCLUSIONS: Using limited echocardiograms for assessing LVH coupled with implementing Canadian technical standards might lead to a greater role for echocardiography in evaluating essential hypertension in certain patients. PMID- 9870125 TI - Defibrillation helps save lives. Heart and Stroke Foundation of Canada. PMID- 9870126 TI - Neoplasms. PMID- 9870127 TI - Biology and genetics of malignant brain tumours. AB - Conventional therapies such as surgery, radiotherapy and, to a lesser extent, chemotherapy have produced significant increases in survival in patients with some types of brain tumours such as medulloblastoma. However, in many other types of brain tumour in both adults and children, the effect of these modalities has been more modest. A thorough understanding of the biology of malignant brain tumours is likely to provide the background for the development of new leads that might be amenable to therapeutic exploitation. This review examines some aspects of glioma biology that have been reported in the past 12 months, and which might be translated into clinical application. PMID- 9870128 TI - Radiation myelopathy. AB - Radiation myelitis has long been recognised as a sinister consequence of spinal irradiation and has limited the acceptable dose of therapeutic radiation to the cord. Over the past 10 years, the pathogenesis has been increasingly understood through the use of animal models. The importance of 'dose per fraction' and 'inter-fraction interval' have been incorporated into new mathematical models which suggest that, for small fractions, the cord may tolerate higher doses of radiation than was previously thought. Clinical recognition of the condition has improved through the description of characteristic magnetic resonance imaging changes. However little advance has been made in its treatment. PMID- 9870124 TI - Coronary artery disease in women. AB - OBJECTIVE: To review and recognize how presentation, investigation, risk factor modification, and treatment of coronary artery disease (CAD) is different for women than for men. QUALITY OF EVIDENCE: Epidemiologic data are from well recognized, peer-reviewed medical journals. Most data on treatment are from randomized controlled trials. MAIN FINDINGS: Coronary artery disease is the leading cause of mortality in women, with incidence after menopause equal to that of men. Diabetes and postmenopausal status without hormone replacement therapy are the strongest risk factors. Women with CAD are more likely to have atypical symptoms, including nonexertional chest pain; pain in other locations, such as jaw, arms, shoulder, back, and epigastrium; and angina-equivalents, such as dyspnea, palpitations, and presyncope. Because women have atypical symptoms, physicians should maintain a high level of suspicion. Although newer nonivasive stress imaging modalities provide greater diagnostic accuracy than traditional exercise stress testing, the tests are still less accurate for women. A safe and cost-effective approach to investigation can be guided by clinical likelihood for CAD based on patients' age, chest pain quality, and risk factors. Treatment and preventive strategies are generally similar for women and men. CONCLUSION: Coronary artery disease is a serious cause of morbidity and mortality in women and will continue to gain importance as women's life expectancy increases. Important differences in presentation, risk factors, investigation, and treatment of women exist and should be recognized. PMID- 9870129 TI - Paraneoplastic syndromes. AB - Paraneoplastic neurological syndromes cause severe neurological dysfunction and may lead to the diagnosis of a potentially curable cancer. The range of clinical presentations of these syndromes continues to grow and new antibody associations are constantly being described. Conventional immunosuppressive therapy is rarely helpful and the prognosis of these conditions remains poor. PMID- 9870130 TI - Driving cognitive and motor gains with rehabilitation after brain and spinal cord injury. PMID- 9870131 TI - Cognitive function outcomes after traumatic brain injury. AB - Investigations of cognitive recovery from traumatic brain injury have utilized experimental cognitive tasks, including measures of executive function and discourse processing, to elucidate specific sequelae. The scope of research, which includes brain imaging and pharmacologic intervention, could potentially enhance rehabilitation of patients with brain injury. PMID- 9870132 TI - Biological interventions for spinal cord injury. AB - Spinal cord injury is frequently followed by the loss of supraspinal control of sensory, autonomus and motor functions at sublesional level. To enhance recovery in patients with spinal cord injuries, three fundamental strategies have been developed in experimental models. These strategies involve three different time points for postlesional intervention in the spinal cord. Neuroprotection soon after injury uses pharmacological tools to reduce the progressive secondary injury processes that follow during the first week after the initial lesion occurs, in order to limit tissue damage. A second strategy, which is initiated shortly after the lesion occurs, aims at promoting axonal regeneration by acting pharmacologically on inhibitors or barriers of regeneration, or by the application of cell or gene therapy as a source of neurotrophic factors or as a bridge or support to enhance the regeneration of lesioned axons. Finally, a mid term substitutive strategy is the management of the sublesional spinal cord by sensorimotor stimulation or the supply of missing key afferents, such as monoaminergic systems. These three strategies are reviewed. Only a combination of these different approaches can provide an optimal basis for potential therapeutic interventions aimed at functional recovery after spinal cord injury. PMID- 9870134 TI - Early detection and treatment of hepatic encephalopathy. PMID- 9870133 TI - Lesion-induced plasticity as a potential mechanism for recovery and rehabilitative training. AB - Brain lesions not only cause a functional deficit in the lesion area, but also affect the structurally intact brain network connected to the lesion. In brain areas surrounding the lesion, as well as those remote from it, the structural and functional plasticity of the brain is increased because of an alteration of transmitter receptor expression and membrane properties of neurones. Within the penumbra of brain ischaemia, as well as after trauma, an additional perilesional dysfunctional zone is found that contributes to the neurological deficit. The lesion-induced plasticity can be used for adaptation, which also may restore function in the perilesional zone, if adequate rehabilitative training is performed. PMID- 9870135 TI - Sex-steroid actions on neurotransmission. AB - Although it is well recognized that sex-steroids exert both developmental and activational influences on the brain, the cellular and molecular mechanisms underlying their actions are less well understood. Progress is rapid, however, and this paper reviews recent insights gained through manipulation of sex-steroid receptor genes, identification of phenotypes expressing these receptors and how these proteins may also be activated to regulate transcription by ligand independent pathways. Advances in our understanding of more rapid actions of sex steroids and the molecular targets involved are also reviewed, as are new studies describing effects on synaptic plasticity and the recent excitement regarding the neuroprotective effects of oestrogen replacement therapy in Alzheimer's disease. PMID- 9870136 TI - Glucose intolerance, cognitive impairment and Alzheimer's disease. AB - The literature on cognitive function and Alzheimer's disease in non-insulin dependent diabetes, impaired glucose tolerance and hyperinsulinaemia are briefly summarized. Neuropsychological, epidemilogical and endocrine contributions are reviewed, with an emphasis on the insulin hypothesis as a potential pathophysiological mechanism for Alzheimer's disease. PMID- 9870137 TI - Neurological aspects of adult phenylketonuria. AB - Phenylketonuria, an autosomal recessively transmitted disorder of amino acid metabolism, is caused by a deficiency of hepatic phenylalanine hydroxylase converting phenylalanine to tyrosine. Thus, phenylalanine accumulates to plasma levels exceeding 1200 mumol/l. Untreated phenylketonuria is characterized by microcephaly, epilepsy, severe mental retardation and, in some cases, progressive supranuclear motor disturbances. These symptoms can largely be prevented by the early start of a phenylalanine-restricted diet. Neurological investigations of treated patients reveal only minor neurological signs, such as tremor or brisk deep tendon reflexes. Magnetic resonance imaging shows white matter abnormalities. However, in single patients, progressive neurological symptoms occurred. Thus, the long-term prognosis of treated phenylketonuria is still under discussion. PMID- 9870139 TI - Neoplasms. PMID- 9870138 TI - Selective vulnerability of the developing brain to lead. AB - Environmental lead exposure in young children who ingest household paint dust or other sources impairs their potential intelligence in a linear, dose-dependent fashion in contrast to its far more subtle effects on other neurologic functions. Basic investigations have identified three interrelated steps in synaptic neurotransmission at which low levels of lead can disrupt signal processing. Lead enhances background transmitter release, but impairs stimulated release, inhibits function at the N-methyl-D-aspartate-type glutamate receptor and stimulates background levels of the intracellular messenger protein kinase C. Taken together these effects have the effect of diminishing the synaptic signal to noise ratio. The ability of lead to enhance 'synaptic noise' during a critical early period of postnatal development may permanently disrupt the architecture of cortical processing units by depriving them of high resolution environmental signals needed to refine synaptic connections. PMID- 9870140 TI - Trauma and rehabilitation. PMID- 9870141 TI - Metabolic disorders and neurotoxicology. PMID- 9870142 TI - Coronary artery remodelling and the assessment of stenosis by pathologists. AB - When atherosclerotic plaques develop, the cross-sectional area of the artery at that point often increases to accommodate the plaque without any reduction in lumen size. In consequence the angiogram does not detect a high proportion of atherosclerotic plaques. The increase in size of the artery (compensatory dilatation-arterial remodelling) varies widely in degree between different plaques even in the same artery. Dilatation of a degree to prevent any loss of lumen size is regarded as adequate compensatory dilatation. In contrast, other plaques are associated with no or minimal increase in the vessel cross-sectional area and a reduction in lumen size in present (inadequate compensation). High grade stenosis is in particular associated with a total failure of remodelling. Such plaques may have had a rapid growth phase, out-pacing the ability of the medial smooth muscle cells to undergo a rearrangement. The phenomenon of remodelling has important consequences for pathologists who use the traditional method of comparing the lumen size relative to the cross-sectional area of the vessel at the site of a plaque to measure stenosis. The area of the vessel at this point may be anything up to 60% above its size before the plaque developed. An error is introduced which on average overestimates diameter stenosis by 30% when compared to an angiographic equivalent method in which the lumen size at the lesion is compared to the lumen size at an adjacent segment of artery without a plaque. PMID- 9870143 TI - Utility of cytokeratin subsets for distinguishing poorly differentiated synovial sarcoma from peripheral primitive neuroectodermal tumour. AB - AIMS: Poorly differentiated synovial sarcoma (PDSS) is a round cell sarcoma that may be difficult to distinguish from other round cell sarcomas, such as peripheral primitive neuroectodermal tumour (pPNET), on histological examination alone. Furthermore, these two tumours may show overlapping immunophenotypes, as some cases of PDSS express CD99, and, on the other hand, pPNET may express epithelial markers. The goal of this study was to determine the utility of cytokeratin (CK) subsets in distinguishing between these two lesions. METHODS AND RESULTS: We evaluated 13 cases of pPNET with RT-PCR detected EWS/FLI-1 fusion transcript and 21 cases of PDSS for the immunohistochemical expression of CK7 and CK19. All cases of PDSS had areas of recognizable monophasic or biphasic synovial sarcoma that expressed at least one epithelial marker. Thirteen of 21 (62%) PDSS stained with AE1/AE3. CK7 and 19 were expressed in 11 (52%) and nine (43%) cases, respectively. Although six of 19 (32%) PDSS demonstrated cytoplasmic staining for CD99, none showed a membranous pattern of immunoreactivity. In contrast, 12 of 13 (92%) pPNET showed strong membranous immunoreactivity for CD99. Four tumours (31%) showed focal staining with AE1/AE3, two of which (15%) stained for CK19. CK7 was not detected in any of the pPNETs. CONCLUSIONS: Although AE1/AE3 may be found in up to 31% of cases of pPNET, the expression of CK7 makes this diagnosis less likely. PMID- 9870144 TI - Synovial sarcoma of the pleura and its differentiation from other primary pleural tumours: a clinicopathological and immunohistochemical review of three cases. AB - AIMS: Synovial sarcomas are rare tumours occasionally arising in the pleural cavity, a site where their histological characteristics may be mistaken for those of malignant mesothelioma. We examined three cases of primary pleural synovial sarcoma in order to look for clinicopathological features that may help in distinguishing them from both mesotheliomas and other sarcomas that may arise in the pleura. METHODS AND RESULTS: All three patients were male, aged 42, 28 and 42, respectively, and had no known exposure to asbestos. One biphasic tumour contained neutral mucin in focal epithelial elements that also stained positively for BerEP4 and AUA1. All three tumours showed focal positivity for either keratin or EMA in the sarcomatous elements, and they also stained positively for bcl-2 protein and MIC2 gene product (CD99). CONCLUSIONS: Our results emphasize the importance of being aware of synovial sarcoma as a possible primary pleural malignancy, especially in a young patient with a localized mass. In addition, the presence of bcl-2 protein perhaps represents a useful marker in distinguishing synovial sarcoma, especially monophasic variants, from mesothelioma within a panel of antibodies. PMID- 9870145 TI - Type 1 protein tyrosine kinases in benign and malignant breast lesions. AB - AIMS: To determine their significance, we examined the expression pattern of the four epidermal growth factor receptor (EGFR) family members as well as the phosphotyrosine kinase activity in breast tumour tissues. METHODS AND RESULTS: Fifty-three malignant breast tumours, four breast cancer cell lines, and 10 benign breast tumours were investigated. Fifty-three per cent (28/53) of the malignant tumours expressed EGFR protein, and the majority of these positive tumours were strongly positive. Eighty per cent (8/10) of the benign tumours also expressed EGFR protein, but all in a lower or moderate level. An association between EGFR expression and increasing malignancy grade was found in the group of infiltrating ductal carcinomas. Of the malignant tumours, 35.8% (19/53) expressed c-erbB-2 protein and 17% (9/53) c-erbB-3 protein, while no expression of c-erbB-2 and c-erbB-3 proteins was found in the benign tumours. Contrary to previous reports, we observed c-erbB-4 receptor protein to be less expressed in the malignant breast tumours. The 'normal' breast epithelial cells adjacent to the malignant tumours and the benign tumours demonstrated intensified membrane staining for c-erbB-4, while a number of the malignant tumours demonstrated a weak cytoplasmic staining or were negative. However, several malignant tumours with strong membrane staining for the c-erbB-4 protein were also found. No simple association between the expression of the four receptors and phosphotyrosine kinase activity was found. CONCLUSION: Our study has revealed a complex expression pattern of the EGFR family members in breast tumour cells. While the data about EGFR, c-erbB-2, c-erbB-3 and phosphotyrosine are largely in line with what has been reported, we found the c-erbB-4 protein expression to be decreased in the malignant tumours. PMID- 9870146 TI - p34cdc2 in invasive breast cancer: relationship to DNA content, Ki67 index and c erbB-2 expression. AB - AIMS: One-hundred and eighty-eight cases of human mammary carcinoma were examined immunohistochemically for their expression of Ki67, p34cdc2 and c-erbB-2. DNA image cytometry was performed to evaluate DNA ploidy, Auer type, S-phase fraction (SPF), 5c exceeding rate (5cER) and 2c deviation index (2cDI). METHODS AND RESULTS: One-hundred and sixty-eight cases were invasive ductal carcinomas, 20 were of invasive lobular type. Routinely assessed oestrogen and progesterone receptor scores were available. The results were analysed statistically in comparison to tumour type, histopathological grade, lymph node status, menopausal status, patient age and overall survival. Ki67 (P < 0.002) and c-erbB-2 (P < 0.0001) correlated well with overall survival (P < 0.0008) and grade (P < 0.038) but not with lymph node status and tumour type. p34cdc2 showed a trend towards a positive correlation with Ki67 (P < 0.058) and a significant negative correlation with receptor status (P < 0.008) but with none of the other parameters examined. CONCLUSIONS: No association between the DNA measured parameters (Auer type, SPF, 5cER and 2cDI) and survival was found. Our results suggest that c-erbB-2 and Ki67 are parameters which might, in combination with receptor status, help to define subgroups with different outcomes. PMID- 9870147 TI - Loss of bcl-2 expression in ductal carcinoma in situ of the breast relates to poor histological differentiation and to expression of p53 and c-erbB-2 proteins. AB - AIM: This study (1) investigates the incidence of bcl-2 protein expression in a series of 108 cases of ductal carcinoma in situ (DCIS), including 25 with early invasive carcinoma, and (2) evaluates the relationship of bcl-2 expression to the histological grade of DCIS and to the expression of oestrogen receptor (ER), c erbB-2 and p53 proteins. METHODS AND RESULTS: The expression of bcl-2, oestrogen receptor (ER), c-erbB-2 and p53 proteins was determined immunohistochemically. Cases were regarded as positive for individual antibodies when at least 10% of the DCIS cells showed positive staining. DCIS was graded histologically as well (n = 9), intermediately (n = 24), or poorly differentiated (n = 75). bcl-2 expression was documented in 57 cases (53%) and was strongly associated with the histological grade of DCIS (P < 0.0001). All cases of well-differentiated DCIS were bcl-2 positive and loss of bcl-2 expression was almost exclusively confined to poorly differentiated DCIS lesions. bcl-2 expression was also closely associated with positive ER status (P < 0.0001). Forty-seven of 57 (82%) bcl-2 positive cases were ER positive while 49/51 (96%) bcl-2 negative cases were ER negative. There was a significant inverse correlation between bcl-2 expression and both p53 protein expression (P = 0.0004) and c-erbB-2 expression (P < 0.0001). Nineteen of 24 (79%) p53 positive cases and 38/45 (84%) c-erbB-2 positive cases showed loss of bcl-2. CONCLUSIONS: Loss of bcl-2 expression occurs in poorly differentiated DCIS and is related to negative ER status and to positive p53 and c-erbB-2 status. This pattern of bcl-2 expression and its association with other biological markers in DCIS is similar to that reported in invasive breast carcinoma. PMID- 9870148 TI - Pigmented atypical fibroxanthoma. AB - AIM: The purpose of this report is to call attention to a pigmented variant of atypical fibroxanthoma that resembles malignant melanoma, both clinically and histopathologically. METHODS AND RESULTS: Thirty-eight cases of atypical fibroxanthoma were examined for the presence of pigmented areas. Four such cases were found. Neoplastic cells showed erythrophagocytosis and accumulation of haemosiderin pigment in the cytoplasm. In three cases, immunohistochemical studies using a battery of antibodies were performed. Neoplastic cells were strongly positive for vimentin and weakly positive for CD68, whereas they were negative for melanocytic markers, including S100 protein, HMB45, and the monoclonal antibody NK1-C3 to melanoma-associated antigen. CONCLUSIONS: Pigmented atypical fibroxanthoma represents a poorly recognized variant of the neoplasm that may be easily misinterpreted as malignant melanoma. To the best of our knowledge, this is the first report of erythrophagocytosis in atypical fibroxanthoma. PMID- 9870149 TI - Malignant proliferating trichilemmal tumours: an histopathological and immunohistochemical study of three cases with DNA ploidy and morphometric evaluation. AB - AIMS: Malignant proliferating trichilemmal tumours (MPTT) are rare neoplasms capable of tissue invasion and metastasis, the diagnosis of which is based essentially on histological features. In difficult cases, however, evaluation of additional parameters may be needed to diagnose malignancy. METHODS AND RESULTS: We report three cases of MPTT in which, in addition to the histological features, we have determined the DNA ploidy, nuclear area and proliferative fraction. CD34 immunoreactivity has also been tested. Two cases were aneuploid, and one diploid with increased proliferating index. PCNA immunostaining labelled 40% and 80% of tumour cells in aneuploid tumours and 30% of the diploid neoplasm. In all cases, nuclear area was consistent with large pleomorphic tumour cells. No CD34 immunostaining was detected. CONCLUSIONS: Aneuploidy is common in MPTT, particularly in tumours with a high proliferative fraction. Loss of CD34 immunoreactivity is an additional feature of potential, though limited, value. Therefore, evaluation of the DNA content, proliferation markers and CD34 immunostaining may be helpful in the diagnosis of MPTT. PMID- 9870150 TI - Blastic NK-cell lymphoma expressing terminal deoxynucleotidyl transferase with Homer-Wright type pseudorosettes formation. AB - AIMS: Rosette-forming malignant lymphoma is very rare. We report a blastic NK cell lymphoma expressing terminal deoxynucleotidyl transferase (TdT) with formation of Homer-Wright type pseudorosettes. METHODS AND RESULTS: An 18-year old boy presented with an enlarged inguinal lymph node. Histologically, the nodal architecture of the lymph node was diffusely effaced by small to medium sized monomorphic blastoid lymphoid cells which frequently formed Homer-Wright type pseudorosettes. Immunophenotyping of the tumour using immunohistochemistry and flow cytometry revealed LCA+, CD4+, CD56+, CD43+, TdT+, CD2-, cCD3-, CD8-, CD7-, CD34- and TIA-1-. DNA analysis revealed no gene rearrangement of TCR beta and gamma genes. In situ hybridization for EBER 1 & 2 was negative. No azurophilic granules were found in the Wright stain. Complete remission was achieved with six cycles of chemotherapy with the CHOP regimen. The disease recurred in the paranasal sinuses and bone marrow 2 years later. CONCLUSIONS: Immunophenotypic and genotypic similarities of the present case to those of TdT-negative blastic NK-cell lymphoma suggest that these diseases might be categorized as one entity irrespective of expression of TdT. PMID- 9870151 TI - Altered expression of membrane inhibitors of complement in human gastric epithelium during Helicobacter-associated gastritis. AB - AIMS: Membrane inhibitors of complement are thought to protect bystander cells from complement mediated damage. Expression of these proteins is enhanced in the colonic mucosa of patients with ulcerative colitis. Our aim was to investigate the regulation of complement activity in Helicobacter-associated chronic gastritis. METHODS AND RESULTS: We immunohistochemically assayed expression of three membrane inhibitors of complement, decay accelerating factor (DAF; CD55), 20 kDa homologous restriction factor (HRF20; CD59) and membrane cofactor protein (MCP; CD46), in 55 biopsy specimens of the human gastric mucosa. DAF, expressed in 33 (60%) of biopsy specimens, and HRF20, expressed in 45 (82%) of the samples, were located mainly on the apical surface of the epithelial cells, whereas MCP, expressed in 48 (87%) of the biopsies, was found on the basolateral surface. We found strong correlation between expression of DAF on gastric mucosal epithelium and the severity of mucosal infiltration of neutrophils (rs = 0.875, P < 0.0001) and mononuclear cells (rs = 0.773, P < 0.0001). No significant correlation was observed between HRF20 expression and neutrophil or mononuclear cell infiltration, while there was a weak negative correlation between MCP expression and these cellular infiltrations. When we assayed immunostaining of Helicobacter pylori (H. pylori) in these biopsy specimens, strong correlation with the degree of neutrophil (P < 0.0001) and mononuclear cell (P < 0.0001) infiltration were observed. The expression of DAF and H. pylori infection in these biopsies were also significantly (P < 0.0001) correlated. No correlation between HRF20 expression and H. pylori infection was observed, but we did find a significant negative correlation (P < 0.005) between the expression of MCP and H. pylori infection. CONCLUSIONS: Our results, demonstrating altered expression of membrane inhibitors of complement in gastric mucosa during inflammation and/or H. pylori infection, suggest that complement may significantly participate in the pathology of gastric inflammation. Moreover, DAF and MCP may play an important role in the regulation of complement activation in the alimentary tract. PMID- 9870152 TI - Expression and subcellular localization of human AP endonuclease 1 (HAP1/Ref-1) protein: a basis for its role in human disease. AB - AIMS: Human AP endonuclease 1 (HAP1) plays a major role in the repair of apurinic/apyrimidinic (AP) sites in cellular DNA by catalysing hydrolytic cleavage of the phosphodiester backbone 5' to the site. HAP1 is also known to be a potent reduction-oxidation (redox) factor, regulating the binding activity of a number of transcription factors. The purpose of the present study was to examine the expression of HAP-1 in a wide range of human tissues. METHODS AND RESULTS: Using a recently developed specific rabbit polyclonal antibody, we performed immunohistochemistry on paraffin-embedded tissue material. Nuclear staining was detected in crypt cells of the small and large intestine, epithelial cells of breast ducts, basal cells of the skin, alveolar cells of the lung, lymphocytes of the marginal zone of the spleen, in the surface epithelium and stromal cells of the ovary and the transitional epithelium of the bladder. Unexpectedly for a presumed nuclear protein, the staining pattern in some cell populations was mainly cytoplasmic (e.g. superficial cells of gastrointestinal tract, Langerhans cells, Leydig cells and spermatocytes, epithelium of the prostate glands), or both cytoplasmic and nuclear (e.g. epithelial cells of thymus, follicular thyroid cells, parietal cells of the stomach, glandular epithelial cells of the cervix, epithelial cells of exocrine pancreas). CONCLUSION: This differential expression in a wide spectrum of cells is indicative of a potential multifunctional action of HAP1, not necessarily restricted to a role in the nucleus. PMID- 9870153 TI - Embolization of mesothelial cells in lymphatics: the route to mesothelial inclusions in lymph nodes? AB - AIMS: To provide evidence that lymphatic embolization is the mechanism for mesothelial inclusions in lymph nodes. METHODS AND RESULTS: A 60-year-old man with alcoholic cirrhosis and ascites had an umbilical hernia resected. The herniorrhaphy specimen contained numerous dermal and submesothelial lymphatic vessels filled by cells similar to the cells that lined the hernia sac. Most of the cells in lymphatics were submesothelial reactive cells, whose cytoplasm stained with antibodies against cytokeratins (AE1-AE3; 8, 18), smooth muscle actin, vimentin, desmin and tissue polypeptide antigen (TPA). Some cells seemed to be superficial mesothelial cells, being positive with high molecular weight anticytokeratin antibody 34 beta E12. On ultrastructural study submesothelial cells with intermediate cytoplasmic filaments, rough endoplasmic reticulum and primitive cell junctions, and scanty superficial mesothelial cells with microvilli, tonofilaments and desmosomes were found in the lymphatics. CONCLUSIONS: Lymphatic dissemination of mesothelial and submesothelial cells is an uncommon and not well known phenomenon. Lymphatic dissemination is probably the route by which the mesothelial cells reach the lymphatic nodes. These cells may be mistaken for malignant cells. PMID- 9870154 TI - Histological demarcation of lateral borders: an unsupportable criterion for distinguishing malignant melanoma from Spitz naevus and compound naevus. PMID- 9870155 TI - Undifferentiated carcinoma of nasopharyngeal type: investigation of Regaud's first case. PMID- 9870156 TI - Myofibroblastoma of the male breast: a comment on differentiation and terminology. PMID- 9870157 TI - Microsatellite instability is not associated with degree of malignancy and p53 expression of gastrointestinal stromal tumours. PMID- 9870158 TI - Hepatic angiomyolipoma with 'oncocyte-like' features. PMID- 9870159 TI - Inflammatory fibroid polyp of the gallbladder. PMID- 9870160 TI - Nodal marginal zone B-cell lymphoma with peculiar follicular colonization involved by malignant lymphoma of thyroid gland. PMID- 9870161 TI - Radiation-associated angiosarcoma involving the parotid gland. PMID- 9870162 TI - Pouchitis: pathophysiology and treatment. AB - Pouchitis is a potential complication after proctocolectomy and restorative ileoanal anastomosis. It is more frequent in UC than in familial polyposis. Little is known about the etiopathology of pouchitis. Risk factors include the presence of extraintestinal manifestations, primary sclerosing cholangitis, cessation of smoking, and previous course of disease. A host of pathophysiological pathways have been identified as potential mechanisms of pouchitis, which include inflammatory mediators, adhesion molecules, oxygen radical species, p-ANCA, and short-chain fatty acids. The microflora in the pouch may also be an important factor in causing inflammation. The risk of developing cancer in cases of pouchitis has not been established as clearly as in those of UC. Particular attention should be paid to patients who have remaining anorectal mucosa after pouch construction. Experience in the treatment of chronic relapsing and chronic refractory pouchitis is limited. The continuation of conventional anti-inflammatory treatment is successful only in a small percentage of patients. New biological response-modifying therapies which target novel immunoregulatory molecules in IBD will also have impact on the systemic and topical treatment of pouchitis. PMID- 9870163 TI - Abnormalities of nerve fibers in the circular muscle of patients with slow transit constipation. AB - Abnormalities of the enteric nervous system are thought to explain the pathophysiology of motility disorders. Our aim was to determine if particular classes of enteric neurons are affected in slow transit constipation (STC). Specimens were taken from the terminal ileum and ascending, transverse and descending colon of patients undergoing subtotal colectomy for STC. Immunohistochemistry was performed using antisera to neuron-specific enolase, tachykinin, leu-enkephalin, choline acetyltransferase, vasoactive intestinal peptide, nitric oxide synthase, tyrosine hydroxylase and neuropeptide Y. The density of nerve fibres labelled with these antibodies in each layer was compared with age-matched controls. The density of nerve fibres with tachykinin and enkephalin immunoreactivity was reduced in the colonic circular muscle of the 15 patients with STC, whereas innervation of all other layers was normal. This reduction of tachykinin-immunoreactive nerve fibres also occurred in nine of the 12 specimens of terminal ileum examined. No difference was detected in the density or distribution of nerve fibres using the other antisera. Excitatory nerve fibres are present in the circular muscle in STC but they are deficient in tachykinins and enkephalin. PMID- 9870164 TI - Simple clinical assessment of colonic response to food. AB - The colonic response to food (CRF) is an integrated function of the colon that has been poorly studied in clinical practice. This study describes a new method to measure it, based on the progress of radio-opaque markers, and shows the results in healthy subjects and in patients with irritable bowel syndrome (IBS). Thirty healthy subjects and 43 patients suffering from IBS were studied. Two studies of colonic transit time (CTT), at rest and after eating a standard test meal, were performed. CRF was quantified by calculating the variation in number of markers in each zone of interest of the large bowel using X-ray films of the abdomen taken before and after eating. The results were as follows: (1) CRF is characterised by caudal propulsion of the colonic contents in the two groups. In controls, there is emptying of the cecum-ascending colon region and filling of the distal large bowel. In IBS patients, only emptying of the left transverse colon and the splenic flexure is found. (2) IBS patients have a delayed CTT because of slowing in the right and left colon, and this is both before and after a meal. The determination of the movements of markers after eating is a simple method that is useful in clinical practice to evaluate CRF. PMID- 9870165 TI - Delorme's operation and sphincteroplasty for rectal prolapse and fecal incontinence. AB - Clinical and manometric results of Delorme's operation and sphincteroplasty were assessed retrospectively in patients undergoing this procedure for fecal incontinence and rectal prolapse. A series of 33 patients (11 males, 22 females; aged 18-83 years, mean 59) with external rectal prolapse were treated by Delorme's operation between 1989 and 1996. Mean follow-up was 39 months (range 7 84). Sphincteroplasty was associated in 12 cases with severe fecal incontinence due to striated muscle defects. Good results were achieved in 27 patients (79%); prolapse recurrence was observed in 6 (21%), the mean recurrence time being 9 months (range 1-24 months). There were no postoperative deaths. Minor complications occurred in 15 patients. Changes in preoperative and postoperative manometric patterns were as follows (mean +/- SEM): voluntary contraction from 59 +/- 6.9 to 66 +/- 7.1 mmHg (P = 0.05), resting tone from 33 +/- 5 to 32 +/- 4.3 mmHg, rectal sensation from 59 +/- 5 to 61 +/- 5.2 ml of air (n.s.). A solitary rectal ulcer syndrome was detected in five patients. The histological pattern demonstrated pathological changes in 40% of cases. Fecal incontinence was resolved in 6 of 20 cases (30%) and chronic constipation in 4 of 9 (44%). Failure (n = 3) was related primarily to postoperative sepsis. The incontinence score showed a mean improvement of 35% decreasing, from 4.5 +/- 0.39 to 2.9 +/- 0.44 after surgery (P < 0.01). In conclusion, Delorme's procedure did not lead to constipation and improved anal continence when associated with sphincteroplasty. PMID- 9870166 TI - Patient satisfaction and symptom relief after anal dilatation. AB - Anal dilatation is used as a simple method of treatment and has been used for both anal fissure and haemorrhoids. This study examined longer-term results among a cohort of 162 patients, 132 of whom responded to a detailed questionnaire, an 82% response (66 patients were male; age range 17-75 years, median 42 years). Follow-up ranged from 16 months to 36 months (median 27 months) after anal dilatation (68 patients for fissure, 32 for haemorrhoids, and 32 for both). In the early months after dilatation, 83% had symptomatic improvement and 76% remained improved. Five (7%) patients with fissure and 11 with haemorrhoids (17%) required further hospital treatment, while 10% and 17%, respectively, had received further treatment from their general practitioners (GPs). Seventy-one percent said they would have a further anal dilation if symptoms recurred. There was no difference in results obtained by surgeons of different seniority. Complications--bleeding (29%) and difficulty controlling flatus (15%) or faeces (8%)--resolved in all cases. The results of anal dilatation for fissure are generally satisfactory in the longer term, with a trend toward better symptom relief in patients with fissure compared with those with haemorrhoids. We do not recommend anal dilatation as the sole treatment of patients with haemorrhoids, but it may be a useful adjunct to other treatments such as banding or sclerotherapy. Morbidity was generally acceptable and the majority of our patients would be prepared to have this procedure again if their symptoms were to return. PMID- 9870167 TI - Intraoperative bowel irrigation improves anastomotic collagen metabolism in the left-sided colonic obstruction but not covering colostomy. AB - This study investigated the effects of intraoperative colonic irrigation and proximal diverting end colostomy after segmental bowel resection in experimental left-colonic obstruction on anastomotic healing. Simple obstruction of descending colon was performed in male Sprague-Dawley rats. After 24 h we performed segmental colonic resection and anastomosis in the control group (n = 15); resection, anastomosis, and covering colostomy in the colostomy group (n = 14); resection and anastomosis after antegrade colonic lavage through cecum by using isotonic saline solution in the irrigation group (n = 13). In rats that were killed 7 days later anastomotic dehiscence and bursting pressure and tissue hydroxyproline concentration at the anastomosis were measured. No significant differences were observed between groups in terms of anastomotic dehiscence, bursting site, or pressure. The hydroxyproline concentration was significantly higher in the irrigation group than the control group (P = 0.025) and the colostomy group (P = 0.029), but no difference was noted between the control group and the colostomy group. These findings suggest that intraoperative antegrade colonic irrigation in the acute left-sided colonic obstruction positively affects collagen metabolism at the anastomotic site; if the anastomosis is performed without bowel cleansing, covering colostomy does not improve collagen metabolism. PMID- 9870168 TI - Histological evaluation of colonic anastomotic healing in the rat following preoperative 5-fluorouracil, fractionated irradiation, and combined treatment. AB - There is a growing interest in neoadjuvant chemo- and radiotherapy as a treatment modality for colorectal cancer which could affect mechanical and biochemical parameters of anastomotic healing. This study investigated the effect of such protocols on colonic anastomotic healing by evaluating the histopathological parameters. One hundred and sixty male Wistar rats were divided into six groups: a control group (I, n = 20), a saline group (II, n = 30) which received 1 ml NaC1 intraperitoneally, a sham-irradiated group (III, n = 20), a 5-fluorouracil (5-FU) group (IV, n = 30), which received 5-FU (20 mg/kg) intraperitoneally for 5 consecutive days, an irradiated group (V, n = 40) which received fractionated irradiation to the whole pelvis to a total dose of 22 Gy, 5.5 Gy per fraction on 4 consecutive days, and a concomitant 5-FU + irradiation group (VI, n = 20) which received 5-FU as in group IV and irradiated as in group V. All groups underwent left colonic resection with primary anastomosis, and the last fraction of irradiation and the last injection were given 4 and 3 days before the operation, respectively. Within each group one half of the animals were killed on the third postoperative day and the other half on the seventh postoperative day. After the resection of the anastomotic segments, histopathological examination was evaluated. Apposition of the wound edges of the mucosa and the muscularis were not affected by the therapy. The level of granulocytes was high, inflammatory exudate and necrosis persisted, granulation tissue formation was delayed, and the levels of macrophages and fibroblasts were low. We conclude that colonic anastomotic healing can be affected by the administration of preoperative chemotherapy, irradiation, and chemoirradiation. PMID- 9870169 TI - Serial evaluation of anorectal function following low anterior resection of the rectum. AB - PURPOSE: This prospective study was performed to serially assess the changes in anorectal function after low anterior resection of the rectum, and to elucidate the mechanisms of functional impairment and the recovery process. MATERIALS AND METHODS: Thirty-two patients undergoing low anterior resection for rectal cancer were evaluated prospectively. Standardized interviews concerning anorectal function and physiologic studies consisting of manometry and balloon proctometry were performed preoperatively, then at 1, 3, and 6 months, and 1 year after the operation. Depending on the length of the residual rectum, patients were divided into two groups: (1) shorter than 4 cm (the short group, n = 18), and (2) longer than or equal to 4 cm (the long group, n = 14). RESULTS: Postoperatively, stool frequency increased and urgency to defecate occurred, which continued until 3-6 months had passed and was more remarkable in the short group. Overall incontinence score increased, which was more remarkable in the short group. Anal resting pressure showed a moderate reduction after 3 months, whereas squeeze pressure did not decrease significantly. Rectoanal inhibitory reflex was postoperatively abolished in almost all patients in the short group, which showed nearly no recovery for 1 year. In the long group, it persisted postoperatively in half the cases, and the reflex returned in a few cases within 1 year. Balloon proctometry revealed overall reduction in rectal capacity and compliance. Although the values tended to recover steadily, they did not reach the preoperative level for 1 year. Urgent volume and maximal tolerable volume remarkably declined, which continued for 1 year and for 6 months, respectively. Rectal compliance also decreased considerably, which continued for 6 months. Most values of rectal capacity tended to be smaller in the short group. CONCLUSION: Impairment of continence after low anterior resection seemed multifactorial, including diminished rectal capacity and compliance, impaired internal anal sphincter tone, and loss of rectoanal inhibitory reflex. Clinical outcome was better and reduction in rectal capacity was less in patients whose rectum remained more than 4 cm. Most of the functional impairments clinically recovered by 6 months postoperation. In the process of clinical recovery of continence, restoration of rectal capacity and compliance and internal anal sphincter tone seemed to contribute a significant degree, while the rectoanal inhibitory reflex did not contribute as much. PMID- 9870170 TI - Radical resection in obstructing colorectal carcinomas. AB - Emergency resections of obstructing colorectal carcinomas usually involve only limited rather than radical lymphadenectomy, which may contribute to the poor long-term survival of these patients. Thirty patients with ileus due to colorectal cancer have been included in a prospective follow-up study since January 1995. Seventeen of these underwent potentially curative resections with radical locoregional lymphadenectomy according to current standards of elective oncological surgery; 2 had radical right and 15 had radical left hemicolectomies. Postoperative morbidity was 18%. An 89-year-old patient died following postoperative bleeding from the colostomy site. During the same period, 13 patients with a metastasizing colorectal carcinoma underwent palliative emergency surgery with a resection rate of only 38%. Morbidity and mortality were 69% and 46%, respectively. These results suggest that emergency radical resections can be safely performed in the majority of patients with obstructing colorectal cancer without increasing the complication rate. PMID- 9870171 TI - Stoma creation for fecal diversion: is the laparoscopic technique appropriate? AB - This study assessed the results of laparoscopic stoma creation for fecal diversion. All patients who underwent elective laparoscopic stoma creation as the sole procedure were evaluated prospectively. Univariate analysis was performed to determine whether previous abdominal surgery, age, gender, body mass index, indication, or surgeons' experience has an effect on the outcome. Between November 1992 and May 1998, 42 patients (17 males, 25 females) with a mean age of 62.1 years (range 17-91) underwent laparoscopic stoma formation. Fecal diversion included loop ileostomy (n = 7), loop sigmoid colostomy (n = 32), and end-sigmoid colostomy (n = 3). Most common indications were unresectable rectal cancer, rectal obstruction caused by advanced pelvic tumors and benign disorders such as perianal Crohn's disease, and fecal incontinence. Of the 42 procedures 41 were completed laparoscopically (97.6%). Complications occurred in four patients (9.5%) requiring reoperation in two (4.8%). Mean length of surgery was 74.4 min (range 30-200). First bowel movements resumed on the 3rd day (range 2nd-7th) after surgery and patients were discharged from hospital after 13 days (range 6 47). Short-term results (> 30 days-1 year) indicated that no further stoma related complications occurred. Analyzing factors potentially predictive of outcome, no statistically significant differences were documented in relation to previous abdominal surgery, age, gender, body mass index, indication, or surgeons' experience (P > 0.05). Laparoscopic stoma creation is appropriate to achieve fecal diversion because it is technically feasible and can be performed with low morbidity. In addition to the benefits of the minimally invasive technique for the patients, laparoscopic stoma formation can be ideal for the surgeon as basic and initial step to perform laparoscopic colorectal procedures. PMID- 9870172 TI - Crohn's colitis: the fate of the rectum. AB - Previous reports suggest that up to 70% of patients undergoing surgery for Crohn's disease of the large bowel do not have gastrointestinal continuity restored and require a permanent ileostomy. In this study the experience with patients requiring surgical treatment of large bowel Crohn's disease is reviewed with particular reference to the management of the rectum. The records of 19 elective and 25 urgent colonic resections performed for large bowel Crohn's disease in 44 patients (16 males, 28 females; mean age 41 years, range 17-76) between 1983 and 1995 were reviewed. Staged proctectomy was performed in 5 of 12 patients who had colectomy for acute colitis and in one patient who had had an elective colectomy. Permanent ileostomy was required in 72% of patients with acute Crohn's colitis and 84% of patients who had elective surgery for large bowel Crohn's. Over 70% of patients having surgical treatment of Crohn's disease of the large bowel required permanent ileostomy. No cases of cancer developed in patients with retained rectal stumps. PMID- 9870173 TI - The interferon-gamma gene as a positional and functional candidate gene for inflammatory bowel disease. AB - Epidemiological and genome-wide linkage analyses have provided firm evidence for a genetic component in the pathogenesis of inflammatory bowel disease. The linkage regions on chromosomes 12 and 16 have been replicated in several independent samples. These represent the best positional evidence in the search for inflammatory bowel disease susceptibility genes. While systematic association and physical mapping studies in these regions are under way, the direct analysis of immunologically relevant genes as positional and functional candidates may provide a shortcut in this process. The interferon-gamma gene resides in the chromosome 12 linkage region near the marker D12S83. Interferon-gamma is an important proinflammatory cytokine in the interleukin-12 cascade and has been implicated in the pathogenesis of mucosal inflammation. We tested this gene for evidence of linkage and association in 133 German multiplex families and 506 single patients with their parents. An intragenic, highly informative CA-repeat marker in intron 1 of the gene was typed using fluorescence-labeled polymerase chain reaction and analysis on an automated sequencer. In the nonparametric linkage analysis using GENEHUNTER, a nonsignificant maximum LOD score of 0.67 was obtained. The transmission disequilibrium test for association was negative (P > or = 0.22) for Crohn's disease, ulcerative colitis, and the combined inflammatory bowel disease phenotype. In summary, the findings make interferon-gamma a very unlikely candidate for the major susceptibility gene in the chromosome 12 linkage interval. Future efforts can concentrate on other transcripts in the region. PMID- 9870174 TI - The penal conceptions of the emperor Marcus Aurelius in respect of lunatics. Reflections on D. 1, 18, 14. PMID- 9870175 TI - Kant on criminal law and psychiatry. PMID- 9870176 TI - Emil Kraepelin and forensic psychiatry. AB - Kraepelin's main positions in forensic psychiatry, that are understandable only in the context of his underlying psychiatric and, especially, nosological theory, are the following: (1) Criminal behavior, especially if repeatedly shown by the same individual, should be regarded as (or, in the strongest version possible, is) mental illness; (2) above all, this viewpoint is due to Kraepelin's broad acceptance of degeneration theory in general, whereas he rejected simplifying concepts like Lombroso's early hypothesis of clinically observable "stigmata degenerations"; (3) Kraepelin voted for the acceptance of diminished responsibility to provide a more differentiated spectrum for the psychiatric expert and the judge; (4) because delinquency, in his view, was closely linked with mental illness, which should be treated instead of (only) being punished, he strictly objected to the death penalty; and (5) he suggested significantly increasing psychiatry's influence on decisions regarding the kind and length of imprisonment. PMID- 9870177 TI - Madness in Florence in the 14th-18th centuries. Judicial inquiry and medical diagnosis, care, and custody. PMID- 9870178 TI - "Dangerous girls," family secrets, and incest law in Italy, 1861-1930. PMID- 9870179 TI - Professional knowledge and professional self-interest. The rise and fall of monomania in 19th-century France. PMID- 9870180 TI - Hysteria, hypnosis, and moral sense in French 19th-century forensic psychiatry. The Eyraud-Bompard case. PMID- 9870181 TI - Criminal lunacy in early modern England. Did gender make a difference? PMID- 9870182 TI - The asylum, the workhouse, and the voice of the insane poor in 19th-century England. PMID- 9870183 TI - Religious mania and criminal nonculpability. Religious and psychiatric reactions to a case of manslaughter in The Netherlands (1900). PMID- 9870184 TI - On-line polymerase chain reaction (PCR) monitoring. AB - In this short note, we present the results of a case study for monitoring the whole polymerase chain reaction (PCR) process (all steps) with a glass fiber fluorometer that was described in a former publication. To utilize this fluorometer, which was originally constructed for a PCR machine with three thermostating devices, a new thermostating device has been developed: the glass fiber matrix is integrated into the thermostating device, while the PCR samples are heated and cooled. The device is able to monitor all samples throughout all stages of PCR with the help of an intercalating dye. This approach also permits one to choose arbitrarily different cooling and heating rates. PMID- 9870185 TI - Increased expression of annexin I and thioredoxin detected by two-dimensional gel electrophoresis of drug resistant human stomach cancer cells. AB - The therapy of advanced cancer using chemotherapy alone or in combination with radiation or hyperthermia yields an overall response rate of about 20-50%. This success is often marred by the development of resistance to cytostatic drugs. Our aim was to study the global analysis of protein expression in the development of chemoresistance in vitro. We therefore used a cell culture model derived from the gastric carcinoma cell line EPG 85-257P. A classical multidrug-resistant subline EPG85-257RDB selected to daunorubicin and an atypical multidrug-resistant cell variant EPG85-257RNOV selected to mitoxantrone, were analysed using two dimensional electrophoresis in immobilized pH-gradients (pH 4.0-8.0) in the first dimension and linear polyacrylamide gels (12%) in the second dimension. After staining with coomassie brilliant blue, image analysis was performed using the PDQuest system. Spots of interest were isolated using preparative two-dimensional electrophoresis and subjected to microsequencing. A total of 241 spots from the EPG85-257RDB-standard and 289 spots from the EPG85-257RNOV-standard could be matched to the EPG85-257P-standard. Microsequencing after enzymatic hydrolysis in gel, mass spectrometric data and sequencing of the peptides after their fractionation using microbore HPLC identified that two proteins annexin I and thioredoxin were overexpressed in chemoresistant cell lines. Annexin I was present in both the classical and the atypical multidrug-resistant cells. Thioredoxin was found to be overexpressed only in the atypical multidrug resistant cell line. PMID- 9870186 TI - New zwitterionic butanesulfonic acids that extend the alkaline range of four families of Good buffers: evaluation for use in biological systems. AB - Four new zwitterionic butanesulfonic acid buffers that are structurally related to four families of Good buffers were evaluated for use in biological systems. These buffers, with pKa values from 7.6 to 10.7, were compared with a variety of other buffers from the same family and with unrelated buffers to determine their effect on enzyme activity and on microbial growth. The activity of four enzymes with optimum pH values in the alkaline range were tested: beta-galactosidase, esterase, phosphodiesterase and alkaline phosphatase. In general, all the Good buffers, including the new butanesulfonic acid buffers, gave good activity; however, there was variation in activity of certain enzymes with certain buffers. Tris, glycine, and phosphate buffers typically showed variation in activity compared to the family of Good buffers. beta-Galactosidase, in particular, showed greater activity with Good buffers than with phosphate or Tris buffers. Similarly, growth of seven bacterial strains was consistent, with a few exceptions, for all the Good family of buffers with Tris often inhibiting growth. Quantitation of alkaline phosphatase conjugated to antibodies is an important tool in many applications in molecular biology. Several Good buffers gave good signals when compared with Tris at pH 9.5 for detection of proteins using alkaline phosphatase-conjugated antibodies. PMID- 9870187 TI - RNA isolation from human skin tissues for colorimetric differential display. AB - High-quality RNA is essential when analyzing expression patterns of tissues by the differential display technique. However, the isolation of intact RNA can be very difficult when tissues are used that contain many RNAses or that are hard to homogenize (e.g. skin samples). We describe an improved protocol for the extraction of high-quality RNA of snap-frozen biopsies from skin tissues that works in a reproducible and reliable manner. In addition, we developed a simplified non-radioactive differential display technique using RNA from small amounts of skin samples. PMID- 9870188 TI - Re-evaluation of the ferrous oxidation in xylenol orange assay for the measurement of plasma lipid hydroperoxides. AB - The ferrous oxidation in xylenol orange version 2 (FOX2) assay coupled with triphenylphosphine has recently been employed for the measurement of total plasma hydroperoxides (ROOHs). In this study, we have evaluated sample handling and the effect of storage conditions on ROOH levels in human plasma (n = 32). Mean level of ROOHs in fresh plasma was 8.35 +/- 3.09 mumol/l (range 4.03-19.5 mumol/l). Addition of butylated hydroxytoluene (BHT) immediately after sample collection had no effect on the concentration of ROOHs. Storage of samples at -70 degrees C for 6 weeks was associated with a variable degree of loss of detectable ROOHs. A mean ROOH level of 6.00 +/- 2.23 mumol/l (range 2.88-13.5 mumol/l) was recorded after 6 weeks of storage at -70 degrees C. There was no difference in the mean level of ROOHs between samples stored for 6 and 60 weeks at -70 degrees C. Inclusion of BHT had no effect on the stability of plasma ROOHs during prolonged storage. Intra-assay coefficients of variation were < 12%, with the lowest variation in fresh samples (7.6%). In conclusion, these results suggest that the FOX2 assay should be a useful tool for measurement of ROOHs in fresh plasma samples but not in stored samples. PMID- 9870189 TI - Polarographic investigation of beta 2-microglobulin and ferritin thermal denaturation. AB - A temperature dependence of the corresponding signals, obtained by differential pulse (d.p.) and alternating current (a.c.) polarography, from a buffered aqueous solution of ferritin and beta 2-microglobulin is used for the characterization of a protein thermal denaturation process. The method is based on the significant differences in the interaction of folded and unfolded protein forms with a dropping mercury electrode due to a different accessibility, for the redox process, of protein electroactive groups. From the analysis of the resulting current, or capacitance, signals in function of temperature the thermal transition reversibility of different protein forms in the solution, protein melting points, and the apparent activation energies of the corresponding processes were determined. PMID- 9870190 TI - Use of Bombyx mori silk fibroin as a substratum for cultivation of animal cells. AB - The growth of animal cells on silk fibroin-coated plates was examined. The anchorage-dependent cells showed almost the same growth on both fibroin- and collagen-coated plates, and it was 30-50% higher than that on polystyrene plates coated with hydrophilic groups. On the other hand, the growth of the anchorage independent hybridomas on the three different plates did not show a significant difference. The cells grown on the respective plates produced their products with the same efficiency despite the difference in the chemical properties of the plates. In this paper, it is demonstrated that silk fibroin can be used as the substratum for the culture of animal cells in place of collagen. PMID- 9870191 TI - Thromboembolic complications following heart valve replacement: the role of patient-related coagulability. PMID- 9870192 TI - High prevalence of hypercoagulable states in patients with recurrent thrombosis of mechanical heart valves. AB - BACKGROUND AND AIMS OF THE STUDY: Thrombosis is one of the most feared and life threatening complications of mechanical heart valves (MHV), with an incidence of 1-3 per 100 patient-years. Hypercoagulable states are highly prevalent in the general population and can predispose MHV to thrombus formation. Thus, we conducted a study to investigate the frequency of hypercoagulable states in patients with MHV who had recurrent thrombosis at least twice after valve implantation. METHODS: Fifteen patients (mean age 42 +/- 11 years; range: 18 to 55 years) with recurrent thrombosis of MHV (2.4 +/- 0.8 recurrences/patient) (group 1) and 15 matched patients (mean age 40 +/- 12 years; range: 18 to 55 years) with MHV without thrombosis (group 2) were followed up with transthoracic and transesophageal echocardiography. Patients' sera were monitored for antibodies to cardiolipin (ACLA-IgG and ACLA-IgM), phosphatidylserine (APSA), lupus-type anticoagulant (LA) and lipoprotein(a) (LP(a)). RESULTS: Average values for group 1 versus group 2 were: ACLA-IgG (normal range < 15 GPLU/ml) 24.7 +/- 14.6 versus 6.2 +/- 2.7 (p < 0.001); ACLA-IgM (< 12.5 MPLU/ml) 7.9 +/- 5.0 versus 3.3 +/- 1.7 (u = 185; p < 0.001); APSA (< 12 RLU/ml) 4.8 +/- 5.7 and 2.9 +/- 1.2 (p = 0.56); and LP(a) (< 30 mg/dl) 36.5 +/- 26.5 and 13.4 +/- 7.1 (p < 0.001). The frequency of LA-positive cases was 4/15 in group 1 and 0/15 in group 2 (p > 0.05). The frequency of abnormally high levels of ACLA-IgG was 9/15 in group 1 and 0/15 in group 2 (p < 0.001); of ACLA-IgM, 2/15 in group 1 and 0/15 in group 2 (p > 0.05); of APSA, 1/15 in group 1 and 0/15 in group 2 (p > 0.05); and of LP(a), 5/15 in group 1 and 0/15 in group 2 (p < 0.05). At least one of the factors included in this study was abnormal in 14 of 15 (93%) patients (p < 0.0001). CONCLUSIONS: Hypercoagulable states are highly prevalent in patients with recurrent thrombosis of MHV. All patients evaluated for therapy of obstructive thrombosis of MHV should be investigated for hypercoagulable state. Moreover, in high-risk patients, surgical replacement of the MHV with a bioprosthesis should be considered. PMID- 9870194 TI - Short-term follow up of the Ross operation in children. AB - BACKGROUND AND AIMS OF THE STUDY: Aortic valve disease in the pediatric population poses special problems to surgeons and cardiologists. The pulmonary autograft has proven to be a good alternative for aortic valve replacement and left ventricular outflow tract (LVOT) reconstruction in this special group. METHODS: Forty-one children (mean age 10.0 +/- 4.8 (SD) years; range: 35 days to 18.8 years) underwent aortic root replacement with a pulmonary autograft between February 1994 and April 1998. Twenty-one patients (51%) had previous cardiac surgery; seven (17%) had balloon valvulotomy. Aortic root replacement was combined with other techniques for various disorders, including tunnel LVOT obstruction, ventricular septal defect (VSD)-aortic insufficiency complex, neoaortic insufficiency following arterial switch procedure, and subvalvular stenosis following correction of type B interruption of the aortic arch (IAA) with VSD (IAA-B/VSD). RESULTS: The mean follow up was 1.7 +/- 1.0 years (range 44 days to 4.1 years). Total follow up time was 67.8 patient-years. Two patients, both after repair of interrupted aortic arch, died intraoperatively (4.9%). There was no late mortality. Two patients were reoperated on (5.1%), one for autograft insufficiency due to cuspal perforation and one for right ventricular outflow tract stenosis at the distal anastomosis. Thirty-eight patients (97%) are currently in NYHA class I; one child with a preoperatively poor left ventricular function did not improve and is in class II. At the latest echocardiographic follow up, neoaortic regurgitation was absent in 19% of patients, trivial in 69% and mild in 11%. Homograft insufficiency was absent in 64%, trivial in 31% and mild in 6%. All mean gradients for both autograft and homograft were < 15 mmHg. CONCLUSIONS: The Ross operation can be performed with good results in infants and children with different forms of LVOT obstruction and aortic insufficiency, though aortic stenosis following IAA-B/VSD repair poses a surgically difficult problem. PMID- 9870193 TI - Prevention of thromboembolism with ticlopidine shortly after valve repair or replacement with a bioprosthesis. AB - BACKGROUND AND AIMS OF THE STUDY: This study investigated the efficacy of postoperative ticlopidine as antiplatelet therapy in patients shortly after heart valve repair or replacement. METHODS: Between 1990 and 1995, 235 consecutive patients underwent either valve repair (n = 67) or replacement with a bioprosthesis (n = 168). The bioprostheses used were Carpentier-Edwards porcine or pericardial (n = 158) valves, Prima stentless valves (n = 3) and cryopreserved homografts (n = 7). Types of repair were aortic (one), mitral annuloplasty with Carpentier ring (65) and tricuspid repair (one). Mean patient age was 67 (range: 16 to 83) years for valve replacement and 57 (range: 32 to 74) years for repair (p < 0.01). Atrial fibrillation occurred in 34% of patients. The hospital mortality rate was 11% (26 patients). Of the 209 survivors, 137 were assigned to antiplatelet treatment with ticlopidine for the first three months of follow up. The other 72 received either oral anticoagulation (coumadin; n = 40), aspirin (n = 14) or no medication (n = 18). In 15 patients, ticlopidine treatment was interrupted due to diarrhea (13 cases), mild allergic reaction (one) or anemia (one). The mean follow up was 3.2 years (range: 1 month to 6 years); cumulative follow up was 684 patient-years (pt-yr) and was complete in 96% of cases. RESULTS: There were two episodes of thromboembolism in the ticlopidine group at 1 month and 6 months respectively, with a linearized incidence of 0.5% pt-yr. In the coumadin group there were four episodes of thromboembolism, three within the first three months of follow up. The linearized incidence was 3% pt-yr (p < 0.01). There were three episodes of hemorrhage in the ticlopidine group in the first three months of follow up and one in the coumadin group. The linearized incidence was 0.75% pt-yr. CONCLUSIONS: Following heart valve repair or replacement with a bioprosthesis, the first three months is a high-risk period for thromboembolism. Ticlopidine seems to prevent this complication better than conventional therapy with oral anticoagulants. Nevertheless, hemorrhage continues to be a problem with ticlopidine therapy. PMID- 9870195 TI - Clinical performance of the native pulmonary valve in the systemic circulation. AB - BACKGROUND AND AIM OF THE STUDY: In a number of corrective and palliative procedures the autologous pulmonary valve is used as the systemic semilunar valve. This study reviews the surgical results and function of the native pulmonary valve in the systemic position after various surgical procedures. METHODS: Between January 1994 and December 1997, the autologous pulmonary valve was transferred functionally or anatomically into the systemic circulation in 89 patients. Follow up echocardiograms and cardiac angiograms were reviewed for 51 neonates with transposition of the great arteries after an arterial switch operation (ASO), in 21 patients after first-stage palliation of hypoplastic left heart syndrome (HLHS), in eight children and adults with pulmonary autograft aortic valve replacement (Ross procedure), and in nine patients with a pulmonary artery-to-aortic anastomosis (Damus-Kaye-Stansel (DKS) procedure) in complex heart defects with outflow obstruction. RESULTS: Nine patients (five with HLHS) died; thus, overall mortality rate was 10.2%. There was no evidence of valve related mortality. Trivial insufficiency following ASO was noted in 11 patients, with no progression of incompetence over time. None of the HLHS patients had pulmonary insufficiency preoperatively, but all showed mild regurgitation on postoperative echocardiography. There was a moderate increase in insufficiency which was attenuated after an early second-stage palliation. Three of nine patients undergoing a DKS anastomosis demonstrated a hemodynamically insignificant insufficiency. Modification of the surgical technique avoided postoperative regurgitation. Four of seven patients having a Ross procedure showed trivial but non-progressive neoaortic regurgitation. CONCLUSIONS: Based on this experience, the autologous pulmonary valve performs adequately at intermediate term follow up. Postoperatively, trivial regurgitation was a frequent finding but was hemodynamically insignificant. Progression or late development of insufficiency as well as stenosis were rare problems. PMID- 9870196 TI - Pure pyrolytic carbon: preparation and properties of a new material, On-X carbon for mechanical heart valve prostheses. AB - BACKGROUND AND AIM OF THE STUDY: Historically, the pyrolytic carbon used in mechanical prosthetic heart valves contained small amounts of silicon, this being a necessary additive to achieve consistently the hardness required for wear resistance. New processing technology has allowed the deposition of pyrolytic carbon without silicon, while maintaining adequate hardness to ensure wear resistance. METHODS: A parametric study of coating parameters identified the conditions necessary to produce the optimal pure carbon material. RESULTS: In comparison with silicon-alloyed carbon, the pure carbon was found to be about 20% stronger, have a strain-to-failure about 25% higher and have a greater toughness. CONCLUSIONS: The enhanced strength, deformability and toughness of the new carbon permits designers to utilize component shapes and dimensions that could not be manufactured using the silicon-alloyed carbons. Such design features have hemodynamic benefits resulting in valve performance improvements. PMID- 9870197 TI - Evaluation of a novel bioprosthetic heart valve incorporating anticalcification and antimicrobial technology in a sheep model. AB - BACKGROUND AND AIM OF THE STUDY: The St. Jude Medical (SJM) Epic valve has been designed to diminish the risk of prosthetic valve endocarditis by the use of silver-coated polyester fabric, and to inhibit dystrophic calcification by the use of ethanol pretreatment. METHODS: A 20-week juvenile sheep mitral valve implant model was used to determine safety and efficacy of the device, as well as the rate of silver release and degree of dystrophic calcification. The SJM Epic valves were compared with SJM Biocor porcine valves (not ethanol-pretreated, not silver-modified polyester fabric) and Baxter Carpentier-EdwardsR standard valves. RESULTS: Blood concentrations of silver reached a maximum of 40 p.p.b. within 10 days of SJM Epic valve implantation, and were well below toxic levels (300 p.p.b.). Blood silver concentrations returned to baseline within 30 days after surgery. Maximal silver accumulation occurred in the liver (16.75 mg/g dry weight); concentrations in the brain, spleen, kidney and lung were similar to those reported for other silver-modified prosthetic valves. No statistically significant difference was found in calcium content between SJM Epic and Biocor valves. The fibrous response to the sewing cuff was similar among the three valve types. CONCLUSIONS: At all times tested, silver release from the SJM Epic valve led to blood concentrations well below toxic levels. Although calcification in the two SJM valve groups was extremely low, the 20-week sheep model may be insufficiently sensitive to detect differences in calcium accumulation in modern bioprosthetic valves. PMID- 9870199 TI - The Ultracor tilting disc heart valve prosthesis: a seven-year study. AB - BACKGROUND AND AIM OF THE STUDY: The Ultracor heart valve is a recent entry in the evolution of the tilting disc valve. This report summarizes the experience with the Ultracor valve from three European centers. METHODS: Between 1990 and 1996, 446 patients received 499 Ultracor heart valve prostheses in 450 procedures, including 225 (50.0%) aortic, 172 (38.2%) mitral and 49 (10.9%) double valve replacements. An additional four (1.0%) patients had mitral valve replacement (MVR) added to a previous aortic valve replacement (AVR) and were considered double valve replacement (DVR) patients. The total follow up was 751 patient-years (pt-yr) for AVR (mean 3.4), 440 pt-yr for MVR (mean 2.6) and 125 pt yr for DVR (mean 2.4). Nine patients (one AVR, eight MVR) were lost to follow up, which was 98% complete. RESULTS: The actuarial survival rate, including operative mortality rate, at five years was 90% for AVR, 77% for MVR and 82% for DVR. The linearized complication rates (%/year) for AVR, MVR and DVR were: 2.1, 4.0 and 4.0 for late mortality; 0.1, 3.0 and 0.8 for thromboembolism; 0, 0.2 and 0 for thrombosis; 2.0, 1.6 and 1.6 for anticoagulant-related hemorrhage (ACH); 0.3, 0.5 and 1.6 for prosthetic valve endocarditis (PVE); and 0.5, 0.9 and 3.2 for reoperation, respectively. The actuarial rates of freedom from complications at five years were: thromboembolism, 99% for AVR and 88% for MVR; thrombosis, 100% for AVR and 99% for MVR; ACH, 91% for AVR and 94% for MVR; PVE, 99% for AVR and 97% for MVR; reoperation, 98% for AVR and 98% for MVR. No structural failure was observed. CONCLUSIONS: Seven years' experience showed the Ultracor heart valve prosthesis to be comparable with other currently used mechanical heart valves. Continued evaluation of this prosthesis is warranted in order to obtain a more extended clinical follow up. PMID- 9870198 TI - In vivo efficacy of antimicrobial-coated fabric from prosthetic heart valve sewing rings. AB - BACKGROUND AND AIMS OF THE STUDY: Antimicrobial coating of medical devices has recently emerged as a potentially effective method for preventing device-related infections. The objective of this animal study was to examine in vivo the antimicrobial efficacy of prosthetic heart valve sewing ring fabric coated with: (i) silver; (ii) combined minocycline and rifampin (M/R); or (iii) combined chlorhexidine and chloroxylenol (CH/CX). METHODS: A rabbit model of Staphylococcus aureus colonization and infection of subcutaneously implanted fabric of prosthetic heart valve sewing rings was used. Following administration of anesthesia and preoperative antibiotic prophylaxis, 0.5 x 0.5 cm samples of fabric were placed subcutaneously into the back of rabbits. Each rabbit received a total of eight samples: (i) two uncoated; (ii) two silver-coated; (iii) two M/R coated; and (iv) two CH/CX-coated. After injecting a bacterial inoculum of 2 x 10(5) c.f.u. of S. aureus onto each implanted sample, the wounds were sutured. Rabbits were monitored daily for one week, killed and the test fabrics removed and cultured. RESULTS: Rates of device colonization, device-related infection and device-related abscess were similar between the uncoated and silver-coated devices. Devices coated with M/R were less likely to be colonized or cause device related infection when compared with uncoated devices, and less likely to be associated with abscess formation than silver-coated devices. There was a tendency for CH/CX-coated devices to be less colonized than uncoated devices. Only M/R-coated and CH/CX-coated devices produced zones of inhibition in vitro. Implantation of M/R-coated and CH/CX-coated devices in rabbits did not result in detectable systemic concentrations of the antimicrobial coating agents. Colonization of antimicrobial-coated devices was not associated with resistant S. aureus isolates. CONCLUSIONS: These results suggest that silver-coated sewing rings may not prove to be clinically anti-infective. In contrast, antimicrobial coated sewing rings that produce effective zones of inhibition, particularly those coated with M/R, are likely to be clinically protective. PMID- 9870200 TI - A comparison of the cavitation potential of prosthetic heart valves based on valve closing dynamics. AB - BACKGROUND AND AIMS OF THE STUDY: This study compares the cavitation potential of prosthetic heart valves based on valve closing dynamics. METHODS: A laser sweeping technique measured valve closing dynamics (average closing velocity and deceleration) immediately before valve closure. A high-fidelity, piezoelectric pressure transducer was mounted proximal to the mitral valve and measured the high-frequency pressure fluctuations caused by cavitation bubble formation and collapse after valve closure. The band-pass filtered root mean squared (RMS) value of the mitral pressure signal was used as a measure of cavitation intensity. The combination of these two techniques allowed the direct correlation of valve dynamics and cavitation intensity for each valve closure. The effects of three parameters on prosthetic heart valve dynamics and cavitation were examined: valve geometry (Medtronic Hall and Bjork-Shiley Monostrut), occluder material (pyrolytic carbon and Delrin), and gap width between the occluder and housing. A dimensional analysis was also performed to investigate the general form of the relationship between valve dynamics and cavitation intensity. RESULTS: For all of the valves investigated in this study, the RMS pressure increased (signifying an increase in cavitation) as the average closing velocity and deceleration increased. In order to compare the cavitation potential of the valves, the RMS pressure was estimated at specific closing velocities using the linear regression of RMS pressure versus average closing velocity for each valve. The effects of valve geometry, occluder material and gap width were then examined at high valve loading conditions (closing velocity of 4.0 m/s). For both pyrolytic carbon and Delrin, the Medtronic Hall valves had significantly higher RMS pressures than did the Bjork-Shiley Monostrut valves. For a given valve geometry, the pyrolytic carbon occluder had a significantly higher RMS pressure than the Delrin occluder. The valve gap width did not have a significant effect on RMS pressure. The dimensional analysis revealed the general relationship among average closing velocity, occluder material properties and cavitation intensity. CONCLUSIONS: The results presented here contribute to our fundamental understanding of cavitation on mechanical heart valves. PMID- 9870201 TI - Loeffler endomyocarditis: a clinical presentation as right ventricular tumor. AB - A four-year-old girl presented with an apical mass of the right ventricle, pulmonary embolism and hypereosinophilia, and was shown to be infected with Toxocara canis. Cardiac involvement was represented by endomyocardial fibrosis, mainly at the apex of the right ventricle, mural thrombi and tricuspid regurgitation. Hypereosinophilia appears to be the head point at the basis of multiple organ damage in Loeffler endomyocarditis. The disease mechanism proposed is the eosinophilic production of peroxidases, which are cytotoxic when released in large amounts into the blood flow. Treatment is based on etiologic and corticosteroid therapy (prednisolone 1-2 mg/kg/day); surgical therapy is indicated in the end stages of the pathologic process. PMID- 9870202 TI - ACC/AHA Guidelines for the Management of Patients With Valvular Heart Disease. Executive Summary. A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee on Management of Patients With Valvular Heart Disease). PMID- 9870203 TI - Benefits of normalizing plasma phenylalanine: impact on behaviour and health. A case report. AB - An elderly man with mental retardation who had never received dietary treatment for his phenylketonuria was placed on a phenylalanine-restricted diet. Social skills and walking gait improved and a new interest in the objects in his environment developed spontaneously. A 2-year analysis of diet, blood plasma phenylalanine levels and behavioural state indicated that small differences in phenylalanine intake impacted his well-being. Of significant note, leg tremor and spasm that precipitated severe self-injury were only reversible when plasma blood phenylalanine concentrations were titrated to near normal ranges and daily phenylalanine intake was strictly controlled. This case may offer a potential explanation for some of the late treatment failures that have been reported and suggest new avenues to explore in the late treatment of PKU. PMID- 9870204 TI - The success of dietary protein restriction in alkaptonuria patients is age dependent. AB - Alkaptonuria is characterized by an increased urinary excretion of homogentisic acid, pigmentation of cartilage and connective tissues, and ultimately the development of inflammatory arthropathy. Various diets low in protein have been designed to decrease homogentisic acid excretion and to prevent the ochronotic pigmentation and arthritic lesions. However, limited information is available on the long-term beneficial effects of these diets. We reviewed the medical records of 16 patients aged 3-27 years (4 > 18 years) to ascertain the age of diagnosis, growth, development, social behaviour, signs of complications and longitudinal dietary compliance. The diagnosis of alkaptonuria was made at an average age of 1.4 years (2 months-4 years); following the diagnosis all patients were prescribed a diet with a protein content of 1.5 g/kg per day. All patients showed normal growth and development, and no major complications of the disease. Behavioural problems associated with poor dietary compliance emerged as the main problem. Dietary compliance decreased progressively with age. The effect of dietary protein restriction in homogentisic acid excretion was studied by fixing the amounts of protein in the diet at 1 g/kg per day and 3.5-5 g/kg per day during 8 days. Twelve patients, aged 4-27 years, participated in the investigation. Protein restriction resulted in a significantly lower excretion of homogentisic acid in the urine of children younger than 12 years (p < 0.01), whereas this effect was less obvious for adolescent and adult patients. The results suggest that restriction of protein intake may have a beneficial effect on alkaptonuric children; but continuation of this regimen to older age seems questionable and not practical. PMID- 9870205 TI - One-methyl group metabolism in non-ketotic hyperglycinaemia: mildly elevated cerebrospinal fluid homocysteine levels. AB - Non-ketotic hyperglycinaemia (NKH) is a rare, severe brain disease caused by deficient glycine cleavage enzyme complex activity resulting in elevated glycine concentrations. Recent experience suggests that factors in addition to glycine kinetics are involved in its pathogenesis. The glycine cleavage reaction through the formation of methylenetetrahydrofolate is an important one-methyl group donor. A deficiency in one-methyl group metabolites, in particular of choline, has been hypothesized in NKH. We investigated metabolites involved in one-methyl group metabolism in plasma and CSF of 8 patients with NKH, and monitored the effect of treatment with choline in one patient. Plasma and CSF choline and phosphatidylcholine concentrations were normal, except for a low plasma choline in the single neonate studied. Choline treatment did not change brain choline content, and was not associated with clinical or radiological improvement. Methionine concentrations and, in one-patient, S-adenosylmethionine and 5 methyltetrahydrofolate concentrations were normal in CSF. Homocysteine concentrations in CSF, however, were slightly but consistently elevated in all four patients examined, but cysteine, cysteinylglycine and glutathione were normal. Serine is important in the transfer of one-methyl groups from mitochondria to cytosol. Serine concentrations were normal in plasma and CSF, but dropped to below normal in CSF in three patients on benzoate treatment. These observations add to our understanding of the complex metabolic disturbances in NKH. PMID- 9870206 TI - Does the polymorphism 677C-T of the 5,10-methylenetetrahydrofolate reductase gene contribute to homocysteine-related vascular disease? AB - Whether the 677C-T polymorphism of the methylene tetrahydrofolate reductase (MTHFR) gene acts as a risk factor for homocysteine-related vascular disease remains a matter of debate. Testing for the 677C-T nucleotide substitution and assay of plasma homocysteine were carried out simultaneously in 69 controls and 113 vascular disease patients from the Paris area. The variant gene frequency as well as the variant homozygous genotype frequency were very similar in controls and patients. Conversely, plasma homocysteine levels were substantially higher in patients than in controls. A slight interaction between the 677C-T MTHFR polymorphism and homocysteinaemia was observed in the patient group only, while a negative correlation between fasting homocysteine and plasma folate levels was found in all individuals homozygous for the 677C-T MTHFR genotype, irrespective of vascular disease. These data suggest that the 677C-T MTHFR polymorphism is not a major determinant of the vascular disease but contributes to increased plasma homocysteine concentration in conjunction with low plasma folate levels. PMID- 9870207 TI - Four novel mutations at the cystathionine beta-synthase locus causing homocystinuria. AB - We describe four new mutations in the cystathionine beta-synthase gene: three point mutations localized in exons 3, 9 and 10 and one mutation in exon 12 which results in stop codon. PMID- 9870208 TI - Comparative biochemical studies of ATPases in cells from patients with the T8993G or T8993C mitochondrial DNA mutations. AB - We performed comparative biochemical studies in cultured fibroblast mitochondria from patients with the T8993G or the T8993C point mutations in the ATPase 6 gene of mitochondrial DNA. We found that ATP production was much more severely decreased in cells from patients with the T8993G mutation than in those from patients with the T8993C mutation. Kinetic studies suggest that both mutations affect only the F0 sector of the mitochondrial ATPase complex. We conclude that these two mutations, which result in the substitution of different amino acids at the same site of the ATPase, result in an enzyme with different biochemical characteristics. PMID- 9870210 TI - Mutational analyses in four Japanese families with X-linked liver phosphorylase kinase deficiency type 1. AB - We analysed the gene of the human alpha-subunit of liver phosphorylase kinase (PHKA2) in four Japanese families with X-linked liver phosphorylase kinase deficiency type 1 by RT-PCR followed by PCR-single-strand conformation polymorphism and direct DNA sequencing. In this study, two novel mutations (Y116D and 2675A-->G) and one mutation previously reported (P1205L) were identified, revealing molecular heterogeneity in Japanese patients. Considering the dissimilarity in phenotype among our patients even with an identical mutation in the PHKA2 gene, it seems that each genetic deficiency in this gene may not be the only factor to determine the clinical heterogeneity in this disease. PMID- 9870209 TI - Biochemical monitoring of the treatment in paediatric patients with mitochondrial disease. AB - Treatment strategies in mitochondrial diseases consist of several drugs that diminish the deleterious effects of the abnormal respiratory chain function, reduce the presence of toxic agents or correct deficiencies in essential cofactors. In this study we evaluated the monitoring of tocopherol, carnitine and ubiquinone concentrations in a group of paediatric patients during a follow-up period of 18 months and the response to treatment of these patients by means of the determination of blood lactate, plasma alanine and oxygen consumption by lymphocytes in relation to the clinical status of the patients. Tocopherol, carnitine and ubiquinone concentrations were easily corrected with therapy. Blood lactate proved the best biochemical tool to assess the response to treatment in paediatric patients. According to our results, improvement or stabilization of the clinical course seems to be more related to the biochemical or molecular defect than to the effectiveness of the treatment. PMID- 9870212 TI - Profound neurological phenotype in a patient presenting with disordered isoleucine and energy metabolism. PMID- 9870213 TI - N-acetylglutamate synthetase deficiency: favourable experience with carbamylglutamate. PMID- 9870211 TI - Cholesterol and oxygenated cholesterol concentrations are markedly elevated in peripheral tissue but not in brain from mice with the Niemann-Pick type C phenotype. AB - Niemann-Pick disease type C (NP-C) is a rare genetic disorder characterized by progressive neurodegeneration, frequent developmental delay and early death. Tissues of affected individuals accumulate large quantities of free cholesterol in lysosomes. Because cytotoxic oxygenated derivatives of cholesterol are known to form readily when cholesterol concentrations are elevated, we searched for these compounds in liver, kidney, spleen and brain from mice with the NP-C phenotype. In order of abundance, we identified 7 alpha- and 7 beta hydroxycholesterol, 5 alpha, 6 alpha-epoxycholestan-3 beta-ol, 4 beta hydroxycholesterol, cholest-4-en-3 beta, 7 alpha-diol and cholest-4-en-3 beta, 6 beta-diol in most tissue samples. Cholesterol concentrations in affected mice were increased 3-fold in kidney and 7- to 8-fold in spleen and liver compared to controls (all p < 0.001) but were unchanged in brain. Although oxysterol levels were markedly elevated in nonbrain tissue, the oxysterol and cholesterol concentrations increased proportionally so that oxysterols expressed as percentage of total sterols were the same for all animals (0.34 +/- 0.19% averaged over all organs in affected animals vs 0.40 +/- 0.42% in control mice). In contrast to peripheral tissue, we could not detect any increase in either absolute or relative oxysterol levels in the brains of affected and control mice (49 +/- 61 vs 53 +/- 43 micrograms/g wet weight and 0.45 +/- 0.52 vs 0.47 +/- 0.37%, respectively). Thus, brain sterols are normal in NP-C mice and it is unlikely that an accumulation of cytotoxic oxygenated derivatives of cholesterol could account for the progressive neuropathology seen in the disease. PMID- 9870214 TI - Unusual pure motor axonal neuropathy in a Burmese family with galactosialidosis. PMID- 9870215 TI - Novel nucleotide substitutions in mitochondrial DNA. PMID- 9870216 TI - Molecular analysis of new Japanese patients with holocarboxylase synthetase deficiency. PMID- 9870217 TI - Psychiatric symptomatology associated with contemporary peacekeeping: an examination of post-mission functioning among peacekeepers in Somalia. AB - Contemporary peacekeepers frequently confront complex stressors including the need to directly enforce peace between warring factions, to deliver humanitarian aid in the midst of political-social devastation, and to balance shifting rules of engagement. As such, it is proposed that participants may be at increased risk for the development of psychiatric distress. The present study examined the types of stressors encountered by 3,461 peacekeepers in Somalia, their current psychiatric functioning as measured by the Brief Symptom Inventory, and the relationship between exposure to various stressors and adjustment. Over one third of participants met criteria for psychiatric caseness. The most commonly reported symptoms included hostility, psychoticism, depression, and paranoid ideation. The best predictors of current functioning were found to be exposure to traditional war-zone-related stressors and general military pride and cohesion. These findings highlight the mental health consequences that service in a peacekeeping mission may have for United States solidiers. Further research is needed to investigate potential mechanisms that could serve as buffers to the stress associated with peacekeeping service. PMID- 9870218 TI - Cognitive coping: the psychological significance of knowing what happened in the traumatic event. AB - The longitudinal relationships between aspects of cognitive coping and psychological symptoms in a group of 36 adults were examined 1, 3, and 5 years after a fatal school bus accident in Norway in 1988. The need for information about what happened in the accident was unrelated to symptoms during the period when the parents' search for facts was dominant, indicating that this is part of a normal adjustment process. However, persistent need for information after factual information had been provided was associated with poorer adjustment. People who felt they were adequately informed and who viewed the accident as a random incident showed better psychological adjustment. PMID- 9870219 TI - Prevalence of traumatic events and peritraumatic predictors of posttraumatic stress symptoms in a nonclinical sample of college students. AB - This study investigated lifetime prevalence of traumatic events and posttraumatic stress disorder (PTSD) symptoms among 937 college students. Participants rated their lifetime experiences of traumatic events and, in response to their "most stressful" event, completed measures of objective stressor dimensions, PTSD, and peritraumatic reactions. Approximately 67% of respondents reported at least one traumatic event. An estimated 4% of the full sample (12% of traumatized individuals) met PTSD criteria within the past week. After controlling for vulnerability factors and objective characteristics, peritraumatic reactions remained strongly predictive of PTSD symptoms. Results are discussed with respect to immediate reactions to traumatic events as potential precursors of PTSD symptomatology. PMID- 9870220 TI - Criminal victimization, posttraumatic stress disorder, and comorbid psychopathology among a community sample of women. AB - This paper provides information on the relation between victimization status, crime factors, posttraumatic stress disorder (PTSD), and several other psychological disorders among a community sample of women. Results indicated that victims of crime were more likely than nonvictims to suffer from PTSD, major depressive episode, agoraphobia, obsessive-compulsive disorder, social phobia, and simple phobia. Furthermore, life threat was associated with increased risk of major depression, agoraphobia, obsessive-compulsive disorder, and social phobia. Completed rape was strongly related to almost every disorder assessed, while robbery and burglary were not related to any disorder. When demographics, victimization status, and crime factors were entered hierarchically into multivariate logistic regressions with PTSD in the final step, associations between victimization status, other crime characteristics (e.g., life threat, injury), and non-PTSD Axis I disorders were greatly reduced. This suggests that PTSD may be an important mediating factor in the victimization-psychopathology relation for many disorders. PMID- 9870221 TI - The function of image control in the psychophysiology of posttraumatic stress disorder. AB - The physiological response to trauma-related stimuli of up to one third of participants with posttraumatic stress disorder (PTSD) cannot be discriminated from that of controls. Psychophysiological measures (heart rate and blood pressure) of 22 PTSD and 23 control civilian participants, all exposed to missile attacks during the Gulf War, were recorded while listening to five scripts. The physiological response of PTSD subjects with high image control (IC) was lower than that of PTSD participants with low IC and similar to that of non-PTSD subjects. The physiological response poorly discriminated high IC PTSD participants from controls, but was successful in discriminating low IC PTSD subjects from controls with 91% specificity and 92% sensitivity. Image control is proposed as a function influencing physiological response in PTSD. PMID- 9870222 TI - Brief early psychological interventions following trauma: a systematic review of the literature. AB - A systematic literature search/review was undertaken of brief early psychological interventions following trauma. Only six randomized controlled trials were found, and none of these included group interventions. Of the six trials, two studies associated the intervention with a positive outcome, two demonstrated no difference on outcome between intervention and non-intervention groups, and two showed some negative outcomes in the intervention group. This review suggests that early optimism for brief early psychological interventions including debriefing was misplaced and that there is an urgent need for randomized controlled trials of group debriefing and other early interventions. PMID- 9870223 TI - Degree of somatoform and psychological dissociation in dissociative disorder is correlated with reported trauma. AB - In this study, the prevalence and severity of traumatic experiences as reported by patients with dissociative disorders and with other DSM-IV psychiatric diagnoses were compared. Furthermore, the predictive value of emotional, physical, and sexual trauma with respect to somatoform and psychological dissociation was analyzed. In contrast with comparison patients, dissociative disorder patients reported severe and multifaceted traumatization. Physical and sexual trauma predicted somatoform dissociation, sexual trauma predicted psychological dissociation as well. According to the memories of the dissociative disorder patients, this abuse occurred in an emotionally neglectful and abusive social context. Pathological dissociation was best predicted by early onset of reported intense, chronic and multiple traumatization. Methodological limitations restricting causal inferences between reported trauma and dissociation are discussed. PMID- 9870224 TI - Transgenerational effects of abusive violence on the children of Vietnam combat veterans. AB - This study examined the relationship between participation in abusive violence in Vietnam and behavioral disturbances among children aged 6-16 in the next generation. As part of the National Vietnam Veterans Readjustment Study (NVVRS) detailed data were obtained on a national sample of male veterans who were living in households with children aged 6-16 (N = 257). Interviews with spouses/partners were used to evaluate current family relationships and child behavior. Children of veterans who participated in abusive violence showed more behavioral disturbance than children of other Vietnam veterans even after multivariate analysis was used to adjust for other factors such as PTSD symptoms, combat exposure, and postmilitary family relationships. Participation in abusive violence appears to affect parent-child relationships in a way that adversely influences children living at home. PMID- 9870225 TI - Early childhood trauma and disorders of extreme stress as predictors of treatment outcome with chronic posttraumatic stress disorder. AB - History of early childhood trauma was prevalent and highly correlated with Disorders of Extreme Stress Not Otherwise Specified (DESNOS) in a sample of veterans in inpatient treatment for chronic posttraumatic stress disorder (PTSD). DESNOS predicted reliable change on a variety of measures of psychiatric symptomatology (including PTSD) and psychosocial functioning independently of the effects of PTSD diagnosis and early childhood trauma history. DESNOS also predicted treatment outcome on PTSD and quality of life measures after controlling for the effects of ethnicity, war zone trauma exposure severity, initial level of symptomatic severity or quality of life, Axis I (PTSD and major depression) and Axis II (personality disorder) diagnostic status, and early childhood trauma history. Early childhood trauma was not predictive of outcome. DESNOS appears to play an important role in assessment and treatment planning for psychotherapeutic rehabilitation of chronic PTSD. PMID- 9870227 TI - An alternative interpretation of "intensive" PTSD treatment failures. AB - An evaluation of program failures in the treatment of combat-related posttraumatic stress disorder has led some reviewers to conclude that the focus of treatment should be shifted away from combat trauma and directed toward other problems. A more detailed examination of these programs reveals that they rarely involve the systematic use of the most soundly-validated PTSD treatment, trauma focused therapy. PMID- 9870226 TI - Persistence of combat-related posttraumatic stress symptoms for 75 years. AB - Investigations of the duration of combat-related posttraumatic stress symptoms have focused mainly on survivors of World War II and the Vietnam War, with little attention to surviving veterans of World War I. The authors describe a case in which posttraumatic stress symptoms persisted for 75 years in a World War I combat veteran and increased in frequency toward the end of his life accompanied by advancing dementia and hospitalization. The case illustrates that posttraumatic stress symptoms may be lifelong and exacerbated by various consequences of aging, even if they are not disabling. PMID- 9870228 TI - Atrocities exposure in Vietnam combat veterans with chronic posttraumatic stress disorder: relationship to combat exposure, symptom severity, guilt, and interpersonal violence. AB - Vietnam combat veterans (N = 151) with chronic posttraumatic stress disorder (PTSD) completed measures of atrocities exposure, combat exposure, PTSD symptom severity, guilt and interpersonal violence. PTSD symptom severity, guilt and interpersonal violence rates were similar to previously reported studies that examined treatment seeking combat veterans with PTSD. Controlling for combat exposure, endorsement of atrocities exposure was related to PTSD symptom severity, PTSD B (reexperiencing) symptoms, Global Guilt, Guilt Cognitions, and cognitive subscales of Hindsight-Bias/Responsibility and Wrongdoing. These results are discussed in the context of previous research conducted regarding atrocities exposure and PTSD. PMID- 9870229 TI - Correlates of psychological distress following armed robbery. AB - Although mental health professionals have long been aware of the impact of traumatic events, it was not until 1980 that the term posttraumatic stress disorder (PTSD) was introduced into the DSM-III. Since then, one major goal of research has been to identify factors associated with distress following trauma; as yet, few reliable indicators have emerged. Within the population of armed robbery victims, this is particularly true. The purpose of this study was to investigate possible correlates of posttrauma distress in armed robbery victims, and to assess the overall level of distress within this group. A questionnaire was mailed out of 57 robbery victims, aged 15 to 65, who were recruited as study volunteers via community outreach. Severity of the trauma, vulnerability attributions, and avoidant coping were significantly related to distress level, and victims exhibited a high level of distress. PMID- 9870230 TI - Adaptation and psychometric properties of the German version of the Dissociative Experience Scale. AB - We introduce the 'Fragebogen zu Dissoziativen Symptomen' (FDS), a German adaptation of the Dissociative Experience Scale (DES) which was developed to screen for dissociation within an ICD-10 framework. In addition to the original 28 DES items, the FDS contains 16 items covering dissociative phenomena included in the ICD-10, particularly pseudoneurological conversion symptoms. The psychometric properties of the FDS were studied in 927 clinical and nonclinical subjects from different diagnostic groups and compared to results of American studies. The scale had good test-retest reliability of .88, high internal consistency (split-half = .90, Cronbach's alpha = .94) and good construct validity. These results indicate that the FDS may be a valuable screen for dissociative psychopathology in German-speaking countries. PMID- 9870231 TI - Trail making test performance in Vietnam combat veterans with and without posttraumatic stress disorder. AB - The present study investigated variables associated with performance on the Trail Making Test from the Halstead-Reitan neuropsychological test battery in Vietnam combat veterans. There was a significant difference in performance between veterans with and without PTSD on both parts of the Trail Making Test. In subgroup analyses excluding participants on medications (antianxiety, antidepressant, and cardiac), comorbid diagnoses (history of alcohol or substance abuse, history of major depression and comorbid anxiety disorder) and compensation-seeking status, the group difference on Trails B remained significant. However, subgroup analyses suggested that poorer performance on Trails A was influenced by antianxiety and cardiac medications, as well as heavy combat exposure status. PMID- 9870232 TI - Exposure to duty-related incident stressors in urban firefighters and paramedics. AB - Little is known about the variables that might be associated with posttraumatic stress symptomatology in high-risk occupational groups such as professional firefighters and paramedics. A sample of 173 urban professional firefighter/EMT's and firefighter/paramedics rated and ranked the stressfulness of 33 actual and/or potential duty-related incident stressors. They also reported whether they had experienced each of these incident stressors within the past 6 months and, if they had, to recall on how many occasions within the past 6 months. A principal components analysis of their rescaled incident stressor ratings yielded five components: Catastrophic Injury to Self or Co-worker, Gruesome Victim Incidents, Render Aid to Seriously Injured, Vulnerable Victims, Minor Injury to Self and Death & Dying Exposure. PMID- 9870233 TI - [Analysis of physiological function by antisense oligonucleotide system]. PMID- 9870234 TI - Selective serotonin reuptake inhibitors in pregnancy and lactation. AB - Selective serotonin reuptake inhibitors (SSRIs) have become the agents of first choice in the treatment of depression because of their safe side effect profile. This paper reviews the current literature on the use of SSRIs in pregnancy and lactation concerning their safety. There are human studies that only used fluoxetine in pregnancy, which established its safety. SSRIs are excreted in breast milk, and their long-term effects on the newborn are unknown at this time. The decision to use SSRIs in pregnancy should be made on a case by case basis with active involvement of the patient in the informed consent process during which the risks and benefits are discussed and documented. PMID- 9870235 TI - Listeriosis during pregnancy. AB - Listeriosis is an uncommon infection that has a unique predilection for pregnant women and may result in pregnancy loss. Contaminated food is the usual source of infection, and increased federal surveillance of foodstuffs is the most effective strategy for prevention of disease. Although dramatic epidemics have received the most publicity, more cases of perinatal listeriosis are isolated. If Listeria chorioamnionitis is diagnosed preterm, in contrast to other types of chorioamnionitis, in utero therapy with high-dose penicillin or trimethoprim sulfamethoxazole is possible, and preterm delivery may be avoided. The clinical characteristics of neonatal listeriosis are similar to neonatal Group B Streptococcus sepsis, with early and late onset forms of disease. The epidemiology, diagnosis, and management of Listeria infection in pregnancy are reviewed. PMID- 9870236 TI - Fecal incontinence in women. AB - Pelvic floor disorders are common in women. Most gynecologists are well versed in the management of urinary incontinence and uterovaginal prolapse; however, knowledge of disorders involving the anorectum is often lacking. This review will discuss the issue of anorectal incontinence in women, attempt to provide a logical outline for evaluation and treatment, and offer potential methods of prevention. PMID- 9870237 TI - Selective photocoagulation of placental vessels in twin-twin transfusion syndrome: evolution of a surgical technique. PMID- 9870238 TI - Recollections of aged Armenian survivors of the Ottoman Turkish genocide: resilience through endurance, coping, and life accomplishments. PMID- 9870240 TI - Resilience in child survivors of the Holocaust: toward the concept of resilience. PMID- 9870239 TI - Reflections on the denial of the Armenian Genocide. PMID- 9870241 TI - Group belonging and mourning as factors in resilience in second generation of Holocaust survivors. PMID- 9870242 TI - Trauma and identity through two generations of the Holocaust. AB - In summary, these four second generation women were/are in search of an equilibrium which includes integrating approach-avoidance feelings about their upbringing. They have struggled in various ways, through their own personal styles, with issues of enmeshment with their parents' ordeals, and this has helped to shape their identities. The approach-avoidance dilemma around the suffering of their parents involves finding a way to separate from, yet to include in the meaning of their lives, the suffering of their parents. In these women, empathy preserves the "good" aspect of the parent and in the end allows for separateness to be achieved. An evolution of their approach to the Holocaust occurs throughout their lives and is enhanced by their having children. Creativity and knowledge play an important role in the expression of tormented feelings in both second and third generations. The issues grappled with by the second generation reverberate in their children. This is a group of resolute, serious people who believe in the preciousness of life. They are also thoughtful, empathic youngsters, aware of social and political inequities. These third generation members feel somewhat burdened by the legacy of the Holocaust, inasmuch as they feel obligated to stand up for Jewish identity and be successful in their own lives. Placing the suffering in a larger group context helps the second generation confront the suffering of their parents. This diminishes individual liability so that the suffering does not have to be taken on personally. The cause goes beyond the self and the family. When, as in Sylvia's case, this outlook is not achieved, the struggle against family enmeshment continues. Seeing the suffering in a group context creates a different set of responsibilities, that of allegiance and closeness to the group. It promotes a need to find meaning in the suffering, a need to cope with the sense of identification with group loss. This urge for empathy is accompanied by its opposite, a wish to dissociate, and the need consequently to negotiate boundaries. The struggle for integration of various feelings becomes a part of their identity in both generations. This paper points to the possibility that, as the Holocaust becomes part of a story, a myth, it becomes a guide (albeit perhaps of a demanding one) by which to live life rather than simply a recall of death. The work of memory has been completed, at times embellished. It also includes, however, a continuous wake up call to vulnerability, a sense of burden, a "chronic" sense of the seriousness and preciousness of life. This paper therefore reflects the fact that suffering can be channeled into identity formation, integrated into an articulation of the meaning of life and a philosophy of life. As the third generation's identity becomes intimately intertwined with its origins, a feeling of continuity is developed which provides a sense of affirmation of the group and of the self. It does, however, also include an awareness of the suffering of that group and in the world at large. PMID- 9870243 TI - Long-term effects of the Holocaust: empirical evidence for resilience in the first, second, and third generation. PMID- 9870244 TI - Coping and resilience across generations: Japanese Americans and the World War II internment. PMID- 9870245 TI - Resilience in unaccompanied minors from the north of Somalia. PMID- 9870246 TI - Trauma and resilience during war: a look at the children and humanitarian aid workers of Bosnia. AB - In summary, resilience in both the children and the workers had key aspects which were definable and similar. Both groups required support and connectedness to others, as well as a need to distance themselves from the impact of the suffering. Both groups required the use of similar mechanisms, such as cognitive restructuring, relaxation, and thought-stopping. The activities of "healthy" individuals in both populations were parallel. In both populations, some actions occurred spontaneously and could be bolstered through circumscribed interventions. In other cases, actions and activities to promote resilience of both populations needed to be created. These actions would affect not only the stress experienced by the workers in the field, improving their quality of life and longevity of service, and reducing costs for the agency involved, but also the children who were being serviced by those workers. Truly working on these strategies seemed to make the difference between short-term coping and longer term resilience. Those individuals who were not able to incorporate the resilience-promoting factors may have been able to cope in the short term with the immediate threat, but then had difficulties following the cessation of the armed conflict. Over the course of my subsequent visits to Bosnia I found a higher rate of alcoholism and suicide among teenagers than anticipated. They had apparently survived the most imminent threat and then succumbed to the long-term stress. Some of these problems were due to demobilization from the army of those teenagers who had been in the military. They were now left with no defined role after having had an important one, which had given them considerable power. The demobilization of their fathers caused problems as well. Friction between the parents arose as the father took authority back from the mother who had been in charge of the family in the father's absence. I also found considerable turnover among the humanitarian aid workers. Those who had been able to stay on the job and be effective in the long run were those who had truly internalized resilience strategies. PMID- 9870247 TI - Some concluding thoughts. PMID- 9870248 TI - Post-transplantation lymphoproliferative disease. Introduction. PMID- 9870249 TI - Clinical and pathological features of post-transplant lymphoproliferative disorders (PTLD). PMID- 9870250 TI - The role of immunosuppression in lymphoma formation. PMID- 9870251 TI - The molecular genetics of post-transplantation lymphoproliferative disorders. AB - The post-transplantation lymphoproliferative disorders represent a significant clinical and diagnostic problem. However, these disorders also represent an important biological model for studying the development and progression of lymphoid neoplasia in immune deficiency. Accurate diagnosis and classification of these disorders requires correlative multiparametric analysis of the clinical behavior of the patient with the histopathological features, immunophenotype, clonal composition, and genetic alterations of the lymphoproliferative disorder. Such analyses should also assist in furthering our understanding of the pathogenesis of these disorders. PMID- 9870252 TI - Role of donor versus recipient type Epstein-Barr virus in post-transplant lymphoproliferative disorders. PMID- 9870253 TI - Epstein-Barr virus gene expression in post-transplant lymphoproliferative disorders. PMID- 9870254 TI - Post-transplant lymphoproliferative disease (PTLD): lymphokine production and PTLD. PMID- 9870255 TI - Interferon-alpha and its effects on post-transplant lymphoproliferative disorders. AB - EBV-transformation induces B lymphocytes to secrete high levels of human IL-10. Additionally, EBV contains a gene, BCRF1, that encodes for a protein that shares activity with human IL-10 in vitro. Thus, infection by EBV seems to promote a Th2 environment in the infected host. One may even hypothesize that EBV-derived IL-10 initiates a cascade of events that promotes a Th2 response and suppresses Th1 activity. This is further confirmed by data that suggest elevated concentrations of IL-4, IL-10, and IgE in patients with PTLD. This implies an association between PTLD and an imbalance in the immunoregulatory system with either an excess suppression of Th1 cells and/or an up-regulation of Th2 cells. One could speculate that if the imbalance in the immunoregulatory system is corrected, the patient's own immune system could potentially defend itself against the virus. Clearly, this is the case in those immunocompromised patients with PTLD who respond to just a reduction in their immunosuppression. Unfortunately, this is only beneficial in approximately half of patients with PTLD. Perhaps this is because patients often do not become entirely immunocompetent, either because all of their immunosuppression cannot be discontinued for fear of rejection or because once the above cascade is established the immune system is not capable of easily switching to the Th1 response necessary for combating the virus. Theoretically, IFN-alpha, because of its anti-viral effect, its anti-neoplastic effect and/or possibly by its ability to promote a Th1 response, should be useful in the treatment of PTLD. IFN-alpha modulates the immune system by several mechanisms including: preventing B cells from producing immunoglobulins, reducing IL-6 receptor density, and augmenting the inhibition of IL-4 by IL-12. In vitro studies document its effectiveness against EBV. Unfortunately, the available evidence as to its efficacy in vivo in patients with PTLD is very limited. At present, there are only 16 reported cases in the literature. There are also three cases of BLPD in immunocompromised patients that were all successfully treated with IFN-alpha and the two cases alluded to earlier from Children's Hospital of Pittsburgh (personal communication). Although the numbers are small, the results are promising. Of the 21 patients with BLPD who received IFN-alpha, 15 achieved complete remission. Four others improved and 2 died from BLPD. One of the 4 that improved died 3 months later from a relapse. Thus, there was an overall mortality of 14% (3 of 21) in those who received therapy with IFN-alpha. This is a very heterogeneous group of patients, several of whom had also received additional therapies. Thus, it is impossible to draw definitive conclusions. However, the mortality rate in this group of patients, who had already failed therapy with a reduction in their immunosuppression, compares very favorably to the reported mortality rate of approximately 23-81% in patients with PTLD. This data suggest that a large multi-centered prospective trial comparing IFN-alpha with and without IVIg to other treatment options (i.e., LAK cells) is warranted in those patients with EBV-positive PTLD who fail to respond to a reduction in their immunosuppression. PMID- 9870256 TI - The antiviral prophylaxis of post-transplant lymphoproliferative disorder. PMID- 9870259 TI - Practical synthesis of androgen: the efficient transformation of 17-oxo group to 17 alpha-hydroxy group. AB - The present report describes the improved transformation of the 17-oxo group in 3 beta-acetoxy-5 alpha-androstan-17-one to a 17 alpha-hydroxy group. A mixture of 17 alpha-acetoxy and 16-ene compounds, which are usually produced by the standard synthetic route, were treated with peracetic acid (epoxidation of the 16-ene compound) and then sodium borohydride-sodium hydroxide (reduction-hydrolysis) to give the desired 17 alpha-hydroxy compound in much better yield than that in previous reports. Recrystallization of the crude product with cyclohexane methanol gave the pure compound in 54% yield (total yield from starting ketone). PMID- 9870258 TI - Regulation of the preovulatory gonadotropin surge by endogenous steroids. AB - Estradiol secreted by growing ovarian follicle(s) has been considered classically to be the neural trigger for the preovulatory surge of gonadotropins. The observation that the estradiol-induced gonadotropin surge in ovariectomized rats is of lesser magnitude and duration than that found in the cycling rat at proestrus has resulted in a search for other steroid regulators. Progesterone is a major regulator of the preovulatory gonadotropin surge. It can only act in the presence of an estrogen background, which is necessary for the synthesis of progesterone receptors. In the estrogen-primed ovariectomized rat, progesterone is able to initiate and enhance the gonadotropin surge to the magnitude observed on the day of proestrus and limit it to 1 day. The physiological role of progresterone in the induction of the preovulatory gonadotropin surge has been demonstrated by the attenuation of the progesterone-induced surge and the endogenous proestrus surge by progesterone receptor antagonist RU486 and the progesterone synthesis inhibitor trilostane. The promoter region of the follicle stimulating hormone (FHS)-beta gene contains multiple progesterone response elements and progesterone brings about FSH release as well. The reduction of progesterone in the 5 alpha-position appears to be important for the regulation of progesterone secretion. Corticosteroids appear to play a significant role in the secondary FSH surge on late proestrus and early estrus. PMID- 9870260 TI - Synthesis and biological activities of 2 beta-chloro-, 2 beta-fluoro-, and 2 beta methoxy-1 alpha,25-dihydroxyvitamin D3. AB - Using 1 alpha,2 alpha-oxido-cholesta-5,7-diene-3 beta,25-diol (2) as a starting material, the provitamins of calcitriol with an additional 2 beta-chloro-, 2 beta fluoro-, and 2 beta-methoxy-substituent (3,4,5) are obtained by transdiaxial opening of the oxirane ring with nucleophiles. An efficient irradiation process is described and used for the synthesis of the 2 beta-substituted calcitriols NS2 (2 beta-Cl), NS6 (2 beta-F), and NS7 (2 beta-OCH3). The affinity of these three vitamin D3 derivatives to the vitamin D receptor (VDR) and was determined. These three A-ring derivatives of 1,25(OH)2D3 were further tested for their proliferation-inhibitory and anti-adipogenic activity and gene regulatoric activity in the vitamin D3-sensitive, murine, mesenchymal cell line C3H10T1/2. The VDR-affinity of the 2 beta-chloro derivative, NS2 (2 beta-Cl), was identical to 1,25(OH)2D3 and its vitamin D binding protein (DBP)-affinity was in the range of 1,25(OH)2D3. NS2 inhibited the proliferation of C3H10T1/2(BMP-4)-cells in the presence of fetal calf serum (FCS) 9 times, and, in the absence of FCS, 111 times lower, as compared with 1,25(OH)2D3. The ID50 dose of adipogenesis-inhibition of NS2 was 13 times higher than the ID50 dose of 1,25(OH)2D3. NS6 (2 beta-F) displayed a slightly higher affinity than 1,25(OH)2D3 to the VDR and DBP affinity. The proliferation-inhibitory activity in the presence of FCS was 90 times higher, as compared with 1,25(OH)2D3. In the FCS-free proliferation assay NS6 displayed an inhibitory activity in the range of 1,25(OH)2D3. NS6 showed an 5 times higher potency to inhibit (pre)adipocyte-differentiation in C3H10T1/2(BMP 4)-cells than 1,25(OH)2D3. NS7 (2 beta-OCH3) showed the lowest VDR-affinity and the highest DBP-affinity of the tested substances, as compared with 1,25(OH)2D3 (11 times lower and 35 times higher respectively). Its proliferation-inhibitory activity in the FCS-free medium was 9 times and in the FCS-containing assay 67 times lower in comparison with 1,25(OH)2D3. A 1250 times higher NS7-dose was needed to reach the anti-adipogenic potency of 1,25(OH)2D3. All tested substances displayed a similar ability to activate a vitamin D responsive element-regulated reporter gene compared to 1,25(OH)2D3 (NS2 and NS6: 1.3 times higher activity; NS7: 1,4 times lower activity). PMID- 9870261 TI - New steroidal anti-inflammatory antedrugs bind to macrophage glucocorticoid receptors and inhibit nitric oxide generation. AB - In continuing efforts to synthesize potent, anti-inflammatory steroids devoid of systemic side effects, methyl 9 alpha-fluoro-11 beta,17 alpha,21-trihydroxy-3,20 dioxo-pregna-1,4-diene-16 alpha-carboxylate (FP16CM) and its 21-acetate derivative (FP16CMAc) were recently synthesized and screened in animal models of inflammation. The compounds have now been assessed for high-affinity glucocorticoid receptor binding and glucocorticoid-mediated inhibition of nitric oxide (NO) generation in an in vitro RAW 264.7 macrophage cell culture system. Relative potencies for glucocorticoid receptor binding were 1, 1.7, and 2.4 for prednisone (P) (IC50 = 287 nM), FP16CM, and FP16CMAc, respectively. Concomitant relative potencies for inhibition of NO generation by macrophages stimulated with lipopolysaccharide were 1, 0.92 and 1.9 for P (IC50 = 126 nM), FP16CM, and FP16CMAc, respectively. Collectively, results suggest that the novel antedrugs are active anti-inflammatory agents. The 9 alpha-fluoro and 21-acetate substituent may contribute to enhanced topical potency, increased receptor binding affinity and inhibitory effects on NO generation. Inhibition of vasoactive NO may be one anti-inflammatory action of the steroidal antedrugs in vivo. Collectively, results suggest that these agents may be useful for topical application in allergic/inflammatory diseases. PMID- 9870262 TI - The scope and limitations of the reaction of delta 5-steroids with mercury(II) trifluoroacetate. AB - The effect of the C-3 substituent on the reaction of androst-5-enes with mercury(II) trifluoroacetate in dichloromethane (modified Treibs oxidation) was investigated. 3 beta-Acyloxyandrost-5-en-17-ones gave 3 beta-acyloxy-6 beta hydroxyandrost-4-en-17-ones accompanied by 3 beta-acyloxy-6-chloromercuriandrost 5-en-17-ones. 3 beta-Acetoxy-6 beta-trifluoroacetoxyandrost-4-en-17-one and 3 beta-acetoxy-4 beta-trifluoroacetoxyandrost-5-en-17-one were revealed to be intermediates in the reaction. The formation of the chloromercury steroids indicated participation in the reaction by the solvent. With 3 alpha acetoxyandrost-5-en-17-one as substrate, a complete reversal in the product distribution was observed. 3 beta-Haloandrost-5-en-17-ones gave mainly products that reflected SN1 substitution of the halide. 3 beta-Hydroxy- and 3 beta trifluoroacetoxyandrost-5-en-17-ones were formed. 3 beta-Methoxyandrost-5-en-17 one afforded in nearly identical yields androst-4-ene-3,17-dione, 3 beta-methoxy 6 beta-hydroxyandrost-4-en-17-one, 3 beta-methoxy-6-chloromercuriandrost-5-en-17 one and 6 beta-hydroxyandrost-4-ene-3,17-dione while androst-5-en-17-one yielded 3 beta,6 beta-dihydroxyandrost-4-en-17-one, androst-5-ene-7,17-dione and androst 4-ene-3,17-dione. The effects of solvent and other mercury salts on the reaction were also studied. Treibs oxidation was successful in chloroform, carbon tetrachloride, and dibromomethane, but not in other solvents tested. 3 beta Acetoxy-6-bromomercuriandrost-5-en-17-one was obtained in dibromomethane. Replacement of the reagent by mercury(II) trichloroacetate altered the intermediates formed but not the products. Mercury(II) tribromoacetate was unreactive, however. PMID- 9870257 TI - Adoptive immunotherapy for Epstein-Barr virus-associated lymphoproliferative disorders complicating marrow allografts. PMID- 9870263 TI - Synthesis and binding affinities of new 17 alpha-substituted estradiol-rhenium "n + 1" mixed-ligand and thioether-carbonyl complexes. AB - The development of technetium and rhenium-based radiotracers for the steroid receptor system requires the use of suitable donor groups on the steroid to provide stable binding sites for the metal. Previous approaches have mainly exploited methods involving various N- and S-coordinating chelate systems or organometallic complexes. In this work, we have prepared several novel chelate systems attached to a series of 17 alpha-substituted estradiol derivatives and examined their binding to the estrogen receptor (ER). The neutral "n + 1" mixed ligand and dithioether-carbonyl complexes that we prepared contain the metal in three oxidation states, +5, +3 or +1, attached to a 17 alpha-substituted estradiol derivative through a thiol group, an isocyanide group, or a dithioether unit, respectively. In our preliminary investigations, we used rhenium as a nonradioactive analog of the radionuclide technetium. All complexes synthesized were evaluated in a competitive radiometric receptor binding assay at 0 degree C and 25 degrees C to determine their relative binding affinities (RBA) to the ER (relative to 3,17 beta-estradiol, RBA = 100%). The complexes show binding affinities up to 23.4% at 0 degree C and 14.1% at 25 degrees C. PMID- 9870264 TI - Enzymic and chemical O-methylation of a 4-hydroxyestrone N-acetylcysteine conjugate. AB - 4-Hydroxyestrone N-acetylcysteine conjugate (4-OHE1-2SR) is considered to be an important compound for monitoring the in vivo formation of catechol estrogen quinones, an intermediary in estrogen carcinogenicity. This article describes the selective synthesis of isomeric monomethyl ethers of 4-OHE1-2SR utilizing the formation of a seven-membered ring lactone by dehydration with acetic anhydride. Using these authentic specimens, enzymic and chemical O-methylation were examined. Enzymic O-methylation, using a rat liver cytosolic fraction, of 4-OHE1 2SR gave its 3-methyl ether as the sole product, while preferential O-methylation of 4-hydroxyestrone (4-OHE1) at the C-4 position was confirmed under the same conditions. Methylation of 4-OHE1-2SR with diazomethane gave initially carboxylate methylation, then the corresponding 3-methyl ether almost exclusively, while methylation of 4-OHE1 also gave its 3-methyl ether preferentially. However, much more rapid formation of the 3-methyl ether was observed with 4-OHE1-2SR than with 4-OHE1 itself. These results show that the hydroxy group at the C-3 position of 4-OHE1-2SR is more reactive than that at the C-4 position, both chemically and enzymatically. PMID- 9870265 TI - C19 steroids estrogenic activity in human breast cancer cell lines: importance of dehydroepiandrosterone sulfate at physiological plasma concentration. AB - The estrogenic action of C19 steroids on breast cancer cells was measured by bioluminescence in stably transfected human breast cancer MCF-7 and T47D cell lines with a reporter gene that allows expression of the firefly luciferase enzyme under control of an estrogen regulatory element. The "estrogenic activity" of C19 steroids, such as dehydroepiandrosterone (DHEA) and its sulfate (DHEAS), androst-5-en-3 beta,17 beta-diol, androst-4-en-3,17-dione, dihydrotestosterone, testosterone, and 5 alpha-androstan-3 beta,17 beta-diol was studied. This showed that DHEAS, at concentration observed in physiological conditions (10(-6) M), had a high "estrogen-like effect" in MCF-7 and T47D cell lines. Other C19 steroids, at physiological plasma concentration, alone or together did not have any significant effect on the luciferase activity. Moreover aminoglutethimide, an inhibitor of the aromatase enzyme, in the presence of C19 steroids, partially decreased the luciferase activity. These results suggest that MCF-7 and T47D cell lines could convert DHEAS to estrogen-like compounds by different enzymatic systems. PMID- 9870266 TI - Lessons from transplantation and future perspectives. AB - Organ transplantation is now a routine treatment for a number of chronic kidney, heart, lung, and liver diseases. We have accumulated much knowledge about these treatments in the respective disciplines, and it seems appropriate to reflect on some general "across-the-border" lessons that may be important for medicine as a whole. The natural history of several diseases has been extended; however, we have also learned much about temporary organ replacement, with the possibility of treating and, perhaps, also preventing some diseases in ways that were not possible in the past. This study explores the phenomenon of temporary organ replacement, whereby organs that are in danger in losing their function may recover in quiescence. It raises the question of whether there might be a common, underlying mechanism-such as apoptosis-for some very different diseases. Pharmacological interventions designed to modulate apoptosis are being developed that will hopefully reduce the amount of time needed for organs to recover their function. We have learned some lessons, but are there more possibilities that need to be explored? PMID- 9870267 TI - Intramucosal pH and intestinal mucosal damage in ischemia-reperfusion injury. AB - Small bowel transplantation (SBT) has become an increasingly promising treatment for short bowel syndrome. The evaluation of graft viability after SBT, however, has not been established, except by mucosal biopsy. We monitored intestinal mucosal acidity in order to detect small intestinal ischemia-reperfusion injury. Mongrel dogs were used in this study. After laparotomy, the small bowel was isolated with a vascular pedicle. A tonometer to measure intramucosal pH (pHi) was then positioned in the terminal ileum. The superior mesenteric artery was occluded with or without concomitant superior mesenteric vein occlusion for 60 or 120 min. The value of pHi was determined from laparotomy (baseline) to 12 h after reperfusion. Whole-thickness specimens of the ileum were taken before ischemia, just before reperfusion, and 1 h afterward. Mucosal injury was graded histopathologically. pHi decreased from baseline in relation to the degree of histopathological mucosal injury. There was a significant correlation between histological findings and the change in pHi. We conclude that monitoring intestinal mucosal acidity is a reliable way of determining graft viability after SBT. PMID- 9870268 TI - Combination splitting using both in situ and ex situ techniques in triple split liver transplantation in pigs. AB - In situ splitting of cadaver livers has been reported to reduce cold ischemic damage, to avoid biliary complications, and to result in improved graft survival. In this study, which involved a wider application of split liver transplantation (SLT), we examined the effects of a technique combining both ex situ and in situ splittings in triple SLT in pigs and compared it to ex situ splitting alone. In the combination splitting group, the splitting between the right and left lobes was done in situ with perfusion of the left lobe with cold, lactated Ringer's solution; that between the lateral and medical right lobes was done ex situ in backtable surgery. The time required for in situ splitting was 28 +/- 5 min. The time for backtable surgery and the total ischemia time were significantly shorter in the combination splitting group than that in the ex situ splitting group (P < 0.05). One day after triple SLT, the elevations in both serum AST and LDH in the ex situ splitting group were significantly greater than those in the combination splitting group (P < 0.05). We conclude that combination splitting may provide a technical improvement and have a beneficial effect on the clinical application of triple SLT. PMID- 9870269 TI - Mild chronic anemia following heart transplantation: a syndrome with prognostic relevance? AB - The clinical relevance of mild chronic anemia in patients after heart transplantation (HTX) has not yet been demonstrated. Forty-five outpatients who had undergone HTX 2-99 months prior to investigation and who had not received blood transfusions or erythropoietin (EPO) before data acquisition were observed over a period of 37 months. Anemia was found in 36 of the 45 patients and was normocytic, normochromic, and slightly anisocytotic (coefficient of variation = 16 +/- 2, normal 11.5-14.5). Anemic patients showed elevated EPO levels, whereas in nonanemic patients EPO levels were normal. Survival after HTX differed significantly in anemic and nonanemic patients (P < 0.02), with 100% survival in the nonanemic and 85% in the anemic group. Chronic anemia in patients after HTX shows a typical pattern. Even when mild, anemia in patients after HTX seems to be of prognostic value and thus might be an indicator of chronic disorders. PMID- 9870270 TI - Attitudes towards organ donation and transplantation--a study involving Baltic physicians. AB - The aim of this study was to identify and describe attitudes towards organ donation and transplantation among a group of Baltic physicians who are involved in this aspect of medical care. A total of 151 neurosurgeons, anesthesiologists, and neurologists anonymously answered a questionnaire between February and March 1995. The majority of physicians said they would be willing to donate their own organs after their death but disagreed with the idea of using organs from a dead person who had had a negative opinion towards organ donation. Given a patient who fulfilled the criteria for brain death, this group of physicians found it widely acceptable to keep the patient on a ventilator until organ donation could take place. We conclude that the participating physicians generally have similar attitudes towards organ donation and transplantation. This study is valuable in that it shows the interest in, and the need for, clinical and research collaboration, including a discussion of ethics, within the area of organ donation and transplantation. This is essential for future collaboration with Western countries. PMID- 9870271 TI - Liver conditioning after cardiac arrest: the use of normothermic recirculation in an experimental animal model. AB - The aim of this study was to compare the possible role of normothermic recirculation with the role of liver transplants from non-heart-beating donor pigs after 20 min of cardiac arrest. Three groups were studied, of which two were control groups: group 1, in which the liver was harvested from a heart-beating donor; group 2, in which the liver was harvested after a period of cardiac arrest followed by total body cooling; and group 3, in which the liver was procured as in group 2, but including a period of 30 min of cardiopulmonary bypass and tissue oxygenation at 37 degrees C before total body cooling. Survival at 5 days; endothelial (hyaluronic acid) and hepatocellular damage (AST, ALT, and alpha GST); adenine nucleotides (energy charge), and histological changes were evaluated. Normothermic recirculation during 30 min showed a significant effect on survival (p = .03), endothelial damage (p < .05), and histological changes after reperfusion (p = .04). Cardiopulmonary bypass significantly increased the energy charge during the normothermic recirculation period (p = .001). Moreover, this study shows that a significant survival (100%) can be achieved with a liver allograft after 20 min of cardiac arrest. Although the liver suffers a major insult in terms of endothelial damage and hepatocellular damage, lesions caused by the ischemic injury are reversible. Histological changes also indicate lesion reversibility, since they almost disappear after 5 days. PMID- 9870272 TI - Laryngectomees' views on laryngeal transplantation. AB - Loss of a functional larynx has marked implications for quality of life that remain after both laryngectomy and its alternatives. One solution is laryngeal transplantation. We hypothesised that laryngeal transplantation would be unacceptable to a population of laryngectomees, and that such a lack of acceptability would not be affected by age, sex or time elapsed since operation. In addition, we sought the views of laryngectomees on priorities for research. A questionnaire was developed and mailed to 1000 members of laryngectomee clubs. A total of 372 of 404 responses were suitable for analysis. Seventy-five percent of the respondents said they would accept a transplant under ideal conditions; the number dropped to 58.9% when a stoma was to be retained. Fifty percent would accept a graft even if it did not result in a normal voice. A positive response was more likely in younger respondents (P < 0.001 all questions; linear regression). Some 47.3% of respondents thought research money could be better spent on other projects, and this response was commoner in older respondents (P = 0.0001). Highest priorities for research into laryngeal cancer were development of new treatments (63.2%), prevention (60.2%) and optimisation of quality of life (57%). In short, there appears to be a surprisingly high level of support for laryngeal transplantation amongst those who have had a laryngectomy. PMID- 9870273 TI - Campylobacter jejuni bacteremia and Guillain-Barre syndrome in a renal transplant recipient. AB - In patients who have not undergone transplantation, Guillain-Barre syndrome (GBS) is typically preceded by an acute infection often sustained by Campylobacter jejuni. Thus far, in renal transplant recipients, only eight cases of GBS have been reported. In seven patients GBS was attributed to cytomegalovirus infection and in the eighth patient to cyclosporin A neurotoxicity. We report here the case of a GBS in a renal transplant recipient following C. jejuni bacteremia. The infection quickly disappeared after erythromycin and methronidazole therapy. GBS progressively evolved into a paraparesis within 1 week. After reaching a plateau phase, the clinical status improved and the patient was able to walk unassisted after 3 weeks. At this last check-up, 54 months later, the patient was doing well with a functioning graft and only minimal weakness of the lower limbs. PMID- 9870274 TI - Circulating interleukin-1 receptor antagonist (IL-1RA) serum levels in patients undergoing orthotopic heart transplantation. AB - In a pilot study we determined the serum levels of circulating interleukin-1 receptor antagonist (IL-1ra) in patients undergoing orthotopic heart transplantation and in control patients scheduled for open heart surgery without allograft transplantation. Blood samples were obtained from 12 transplant recipients and 7 controls prior to the operative procedures to determine baseline values. Serum levels of IL-1ra were measured within 12 h of decrossclamping of the aorta and every 24 h for the following 14 days. Endomyocardial biopsies were obtained weekly for the 1st month after transplantation. Compared to baseline values, IL-1ra serum levels 12 h after decrossclamping of the aorta were significantly higher both in the control group (507 +/- 165 vs 3980 +/- 452 pg/ml, P < 0.01) and among the transplant recipients (413 +/- 180 vs 4117 +/- 459 pg/ml, P < 0.01) IL-1ra levels remained significantly elevated for 2 and 5 days, respectively. There were no significant differences in the IL-1ra serum levels between the two groups throughout the observation period. Endomyocardial biopsies of two patients showed acute allograft rejection, Billingham grade IIIa and IIIb, respectively. In both cases, the rejection episodes were accompanied by a renewed and more pronounced elevation in the IL-1ra serum levels beyond 4000 pg/ml for at least 2 days. These preliminary results indicate that IL-1ra may be a nonspecific immune marker during the first few days after orthotopic heart transplantation and cardiopulmonary bypass. Moreover, renewed, prolonged increases in IL-1ra appear to be associated with rejection. Further studies are needed to confirm the predictive value of IL-1ra in the detection of acute allograft rejection. PMID- 9870275 TI - Laparoscopic cholecystectomy injury: an unusual indication for liver transplantation. AB - The introduction of laparoscopic cholecystectomy has been associated with a rise in the number of reported bile duct injuries (0.3%-0.8%). Significant vascular injuries are rare (0.16%), but may lead to life-threatening complications. We present a case report of a patient undergoing transplantation for a laparoscopic cholecystectomy injury. PMID- 9870276 TI - Rehabilitation of long-term defunctionalized bladder for renal transplantation. AB - Some kidney transplant candidates have urinary diversion due to lower urinary tract dysfunction. Careful assessment of the native defunctionalized bladder before kidney transplantation may reveal a usable bladder in a substantial number of these patients. However, most of these bladders need some kind of rehabilitation. An assessment of the continence mechanism is also crucial. We report on successful kidney transplantation into a native bladder, defunctionalized for 9 years, after it had been carefully evaluated and rehabilitated. PMID- 9870277 TI - Firstborn right OR a son with more than one father? PMID- 9870278 TI - Expanded criteria donors and dual kidney transplantation. PMID- 9870279 TI - Minimally invasive approaches in urology. PMID- 9870280 TI - A novel method of surgical instruction: international telementoring. AB - Telemedicine is the use of communication technologies to deliver health care. Telesurgical telementoring represents an advanced form of telemedicine, whereby an experienced surgeon can guide and teach practicing surgeons new operative techniques utilizing current video technology, medical robots, and high-band width telecommunications. This technology can potentially enhance surgeons' education, increase patients' access to experienced surgeons, and decrease the likelihood of complications due to inexperience with new techniques. A personal computer-based system was developed to immerse a surgical specialist into a distant operating room utilizing public telephone lines. Telesurgical laparoscopic telementoring has successfully been implemented in 27 prior operations between the Johns Hopkins Bayview Medical Center and the Johns Hopkins Hospital, whereby the two institutions were separated by 3.5 miles. We report our experience in performing three successful international surgical telementoring operations, one in Innsbruck, Austria, and two in Bangkok, Thailand. PMID- 9870281 TI - The "mini-perc" technique: a less invasive alternative to percutaneous nephrolithotomy. AB - The disadvantages of standard percutaneous nephrolithotomy (PCNL) as compared with ureteroscopy or extracorporeal shock-wave lithotripsy include increased blood loss, greater pain, and longer hospital stay. A 13-Fr "mini-perc" technique using a ureteroscopy sheath for PCNL was developed in an attempt to address these drawbacks. Nine "mini-percs" have been performed in patients aged 40-73 years with stone burdens of < or = 2 cm2. On average, patients had 1.4 stones with a cross-sectional area of 1.5 cm2. The mean total procedure time, estimated blood loss, and hematocrit decrease were 176 min, 83 ml, and 6.6%, respectively. On average, patients used 14 mg of parenteral morphine and stayed 1.7 days in the hospital. There was no procedure-related complication or transfusion. Eight of nine kidneys (89%) were stone-free on early follow-up at a mean of 3.8 weeks. As compared with standard PCNL, the "mini-perc" technique has similar early success rates in selected patients and may offer advantages with respect to hemorrhage, postoperative pain, and shortened hospital stays. PMID- 9870282 TI - Safety and efficacy of tubeless percutaneous nephrostolithotomy. AB - The experience at this institution with tubeless percutaneous nephrostolithotomy was reviewed to determine its safety and efficacy. Between April 1997 and June 1998, 33 "tubeless" percutaneous nephrostolithotomies were performed. All procedures were performed by a single surgeon (R.W.W). All patients had an internal ureteral stent placed at the time of surgery, and a Foley catheter remained in place overnight. All patients had their nephrostomy tube removed in the OR at the end of their surgery. The length of hospitalization, operative time, patients' comorbid conditions, pre- and postoperative hematocrits, transfusion requirements, reasons for a stay of longer than 24 h, complications, stone burdens treated, residual stone disease, any additional procedures required, and postoperative analgesia requirements were reviewed. All 33 percutaneous procedures were performed without significant complication. No transfusion was required. The average length of hospital stay was 1.5 days, with two-thirds of patients staying less than 24 h. A 94% stone-free rate was achieved, and patients had minimal analgesia requirements. This experience with the "tubeless" percutaneous nephrostolithotomy indicates that it is a safe and effective means of stone management. It will likely have an expanding role in the treatment of stone disease and other urologic problems of the upper urinary tract. PMID- 9870283 TI - The prostate anesthetic block for outpatient prostate surgery. AB - With the emergence of minimally invasive techniques for the treatment of benign prostate hyperplasia and the need to reduce health care costs, a simple and effective prostate anesthetic block may allow more procedures to be done in an outpatient or office setting. As based on neuroanatomy studies of the prostate, the perineal approach appeared to be the best way to anesthetize the prostate. This technique was used in 43 patients who underwent interstitial laser coagulation an outpatient or office setting. The perineal prostatic block was effective for pain control whether lidocaine or lidocaine/bupivacaine was used as determined by visual analog scale, linear pain scale, or global pain questionnaire. The use of intravenous (i.v.) sedation did not influence the efficacy of the prostate anesthetic block. Thus, the perineal prostate anesthetic block may be employed in patients undergoing minimally invasive surgery for benign prostatic hyperplasia in the outpatient/office location. PMID- 9870284 TI - Office-based surgery and anesthesia: where are we now? AB - At the time of publication of this journal, the Center for Urinary Control in Memphis, Tennessee, has performed eight office-based surgical procedures using anesthesia. Patients were aged between 56 and 80 years, and all were ASA I-III class. The procedures involved either cystoscopy with collagen injection into the bladder neck or cystoscopy with indigo laser treatment of the prostate. All patients were prepared by the anesthesiologist prior to the day of surgery, and appropriate laboratory tests were ordered and evaluated. All patients did very well, the surgical field was quiet, and each procedure went very smoothly. Each patient tolerated anesthesia without any adverse effect, was recovered in an adjacent examination room, and was extremely satisfied upon discharge from the office. Anesthesia was induced with Propofol (Diprivan) and sublimaze, maintained with a Propofol infusion and oxygen. The surgeon performed a periprostatic block on the patient having the Indigo laser treatment. PMID- 9870285 TI - The University of Tennessee experience with the Indigo 830e laser device for the minimally invasive treatment of benign prostatic hyperplasia: interim analysis. AB - Benign prostatic hyperplasia (BPH) is the most common benign tumor in men and is responsible for urinary symptoms in the majority of men older than 50 years of age. Although transurethral resection of the prostate (TURP) is the gold standard, its complications have impacted upon its utility. As a consequence, new pharmacologic and minimally invasive approaches to the management of BPH have been developed. One minimally invasive approach that employs interstitial laser coagulation by the Indigo 830e LaserOptic system heats the prostate to the point of irreversible necrosis while preserving the urethral lining, potentially resulting in fewer complications. To test the efficacy of this device we evaluated the interim results obtained in 25 patients treated for BPH. Parameters evaluated included the AUA symptom score, uroflowometry, post-void residual, and prostate size. Following treatment, patients were discharged home and the catheter was removed within 3-7 days. Patients were assessed at 1 month and at subsequent 3-month intervals following the procedure using a questionnaire, AUA symptom score, and uroflowometry. The results of the paired t-tests demonstrated a significant increase in the maximal and average flow rates from baseline. The mean baseline maximal flow rate was 8.3 ml/s and increased to 10, 12.7, 14.1, and 12.0 ml/s at 1, 3, 6, and 9 months, respectively, and the mean baseline average flow rate was 4.4 ml/s and increased to 5.3, 6.0, 6.6, and 6.2 ml/s at 1, 3, 6, and 9 months, respectively. The AUA symptom scores decreased from 20.2 to 9.8 at 9 months. There was no intraoperative complication. Six patients developed transient retention. No patient developed bladder neck contractures, urinary incontinence, impotence, or urinary tract infections. One patient developed retrograde ejaculation and one patient required retreatment by TURP. Hence, improvements in symptom scores and voiding parameters suggest that the laser interstitial coagulation prostatectomy is safe and effective for the treatment of BPH. PMID- 9870286 TI - Interstitial laser coagulation of the prostate: introduction of a volume-based treatment formula with 12-month follow-up. AB - The objective of the present study was to assess the efficacy of interstitial laser coagulation (ILC) of the prostate using a low-volume treatment formula as a minimally invasive therapy for symptomatic benign prostatic hyperplasia (BPH). A total of 25 men underwent ILC of the prostate for symptomatic BPH between February 1997 and June 1997. The Indigo 830e laser system from Indigo Medical Inc. and the factory preset therapy regimen was used for ILC of the prostate. The number of treatments or punctures was determined by the formula 0.5 x total prostate volume (ml)/8 ml, rounded to the closest even whole number. The treatment outcome was evaluated at 3-, 6-, and 12-month intervals using the American Urological Association's (AUA) BPH symptom score, maximal urinary flow, prostate size, and postvoid residual urine volume. The AUA symptom score decreased from 23.2 (range 17-28) prior to treatment to 9.4 (range 4-14) at 3 months, 6.6 (range 5-12) at 6 months, and 7.2 (range 4-11) at 12 months. The maximal flow rate improved from 8.4 (range 5-10) ml/s pretreatment to 14.1 (range 10-20) ml/s at 3 months, 14.8 (range 10-18) ml/s at 6 months, and 16.8 (range 12 25) ml/s at 12 months after treatment. There was no significant postprocedure complication. The 1-year clinical results suggest that the ILC procedure using the Indigo 830e device in conjunction with a low-volume treatment formula has outcomes similar to those obtained following ILC based on high-volume coagulation. PMID- 9870287 TI - Minilaparotomy staging pelvic lymphadenectomy follow-up: a safe alternative to standard and laparoscopic pelvic lymphadenectomy. AB - Minilaparotomy pelvic lymphadenectomy (MINILAP), which utilizes a small 6-cm lower midline incision with the aid of standard retractors and instruments, was introduced in 1992 as an alternative to standard and laparoscopic pelvic lymphadenectomy. Since that time, MINILAP has been popularized by the urology community to become the procedure of choice for pelvic lymphadenectomy in patients who are at high risk for lymph-node prostate-cancer metastasis. A total of 24 patients who underwent MINILAP were followed prospectively for the assessment of perioperative morbidity. For the 15 patients who had MINILAP alone the length of hospitalization was < 1 day and the postoperative analgesic requirement was < 10 tablets of oral pain medication. All patients resumed their normal activity within 3-5 days. No acute or late complication was observed. Hence, MINILAP appears to be a safe alternative to either laparoscopic or standard pelvic lymphadenectomy. PMID- 9870288 TI - Continence mechanism of the ileal neobladder in women: a urodynamics study. AB - We undertook this study to evaluate the mechanism of continence in women who underwent modified radical cystectomy and creation of an ileal neobladder. Our surgical technique was modified in accordance with detailed anatomic dissection of female pelvises with attention to the innervation of the pelvic musculature and urethral sphincter. Ten women aged 41-71 years (mean 64.3 years) underwent nerve-sparing radical cystectomy and creation of an orthotopic neobladder with detubularized ileum. Videourodynamic evaluation was performed 6 months postoperatively to evaluate sphincteric and reservoir function. Seven of the ten patients were totally continent after the procedure, requiring no protective pad. Of these, one requires intermittent self-catheterization. Videourodynamic evaluation revealed a low-pressure reservoir with a mean capacity of 467 ml, and leakage did not occur during Valsalva maneuver. Three patients reported significant incontinence (more than one pad per day) after orthotopic reconstruction. These patients demonstrated intrinsic sphincteric deficiency with a low mean abdominal leak-point pressure of 48.3 cmH2O. Two of these women had stress incontinence preoperatively. In conclusion, continence can be preserved in most women after modified radical cystectomy and orthotopic bladder replacement. Success results from preservation of the intrinsic sphincteric mechanism and the creation of a low-pressure, compliant reservoir. A history of stress incontinence preoperatively appears to predispose to sphincteric weakness postoperatively. PMID- 9870289 TI - Peptidyl membrane-interactive molecules are cytotoxic to prostatic cancer cells in vitro. AB - Cytotoxic membrane disruption via lytic peptides is a well-recognized mechanism of immune surveillance for antifungal and antibacterial host protection. Naturally occurring lytic peptides were shown to exhibit antitumor activity as well. Peptidyl membrane-interactive molecules (MIMs) are synthetic lytic peptides specifically designed to maximize antitumor activity. We tested nine novel Peptidyl MIMs for activity against four androgen-insensitive prostate-cancer cell lines using a standard microculture tetrazolium (MTT) assay. Five Peptidyl MIMs known to form alpha-helical secondary structures were active against prostate carcinoma and were chosen for further study. Three peptides configured in beta pleated sheets were noticeably less effective. Concentrations lethal to 50% of the prostate-cancer cell lines treated (D50 values) with the five chosen Peptidyl MIMs ranged from 0.6 to 1.8 microM. For comparison, two alpha-helically structured peptides, D2A21 and DP1E, were tested on several other cancer types: breast (n = 2), colon (n = 2). bladder, cervical and lung carcinomas (n = 1 each). Resulting LD50 values obtained in breast carcinoma cells were significantly higher (P < 0.05) than those observed in prostate cancer cells. LD50 values recorded for D2A21 and DP1E in cervical, colon, bladder, and lung cancer lines were similar to those obtained in prostate cancer cells. As compared with cisplatin, a standard chemotherapeutic drug, the LD50 values recorded for D2A21 were significantly lower (P < 0.04) in prostate-cancer cell lines, suggesting the therapeutic efficacy of Peptidyl MIMs. These data demonstrate for the first time the cytotoxic potential of Peptidyl MIMs against prostate cancer cells and suggest a dependence on a specific secondary alpha-helical structure of the peptide. PMID- 9870290 TI - An alternative procedure for the creation of an ileal conduit in patients undergoing pelvic exenteration: dextrotransmesenteric location. AB - Ileal conduits have long been accepted as a standard method for urinary diversion, but conventional ileal conduits are not always suitable for patients whose ureters are for the greater part compromised by neoplasm or preoperative irradiation, resulting in a shortening of the ureters. Bowel migration into the large cavity, which develops after pelvic organ removal, appears to cause tension on the ureteroileal anastomotic site. Dextrotransmesenteric placement of an ileal conduit may provide easy access to the shortened ureters without exerting pressure on the anastomosis. We employed this procedure in seven patients undergoing pelvic exenteration with relatively minimal morbidity. PMID- 9870292 TI - Central autonomic innervation of the lower urinary tract--a neuroanatomy study. AB - Knowledge about the central innervation of the lower urinary tract is limited. The spinal cord and the pontine micturition center have been investigated most thoroughly, whereas high centers have received little attention. Pseudorabies virus (PRV), a self-amplifying and transneuronal tracer was injected into the bladder trigone of 21 Sprague-Dawley rats. The animals were killed after 72, 96, and 120 h. The whole CNS was sectioned and immunostained for PRV. CNS centers directly connected to the bladder include the intermedio lateral cell column, the central autonomic nucleus, and the nucleus intercalatus at the spinal cord levels T12-L2 and L6-S2. The raphe pallidus et magnus, the A5 nor-adrenergic area, the pontine micturition center, the locus coeruleus, the periaquaductal gray, the nucleus para- et periventricularis of the hypothalamus, the red nucleus, the medial preoptic area, and the cortex are supraspinal centers connected to the bladder. Lower urinary tract function is a complex multilevel and multineuronal interaction. It involves facilitation and inhibition at many levels of the CNS. PMID- 9870291 TI - N-acetyl-beta-D-glucosaminidase excretion in healthy children and in pediatric patients with urolithiasis. AB - Hyperoxaluria was reported to induce renal damage, probably due to toxic effects on renal tubules. Such tubular damage might be expressed by an increase in urinary excretion of marker enzymes such as N-acetyl-beta-D-glucosaminidase (NAG). We set out to examine a possible relationship between the excretion of NAG and that of urinary lithogenic and stone-inhibitory substances by analyzing 24-h urine specimens from 56 children with urolithiasis and 25 healthy children with normal renal function and without a history of urolithiasis. The NAG excretion was higher in patients with urolithiasis (3.5 +/- 0.51 U/g creatinine) as compared with healthy subjects (1.33 +/- 0.14 U/g creatinine, P < 0.05). A positive correlation between NAG and oxalate excretion was observed in female patients (r = 0.56: P < 0.01). In conclusion, the increase in urinary NAG in children with urolithiasis might express renal tubular damage. It seemed, however, not to be specifically related to the excretion of a single lithogenic substance. PMID- 9870294 TI - Interference from short-term memory processing on encoding and reproducing brief durations. AB - A temporal reproduction task is composed of two temporal estimation phases: encoding of the interval to be reproduced, followed by its reproduction. The effect of short-term memory processing on each of these phases was tested in two experiments. In Exp. 1, a memory set was presented, followed by two successive tones bounding the target interval to be reproduced. During the reproduction of the target interval, a probe was presented, and the subject ended the reproduction by pressing one of two keys, depending on the presence or absence of the probe in the memory set. In Exp. 2, probe recognition was required during the encoding of the interval to be reproduced. Whereas in Exp. 1 reproductions lengthened as a function of memory-set size, in Exp. 2 temporal reproductions decreased with set size. These results support attentional models of time estimation and suggest that short-term memory processing interrupts concurrent accumulation of temporal information. PMID- 9870293 TI - Sensory effects on judgments of short time-intervals. AB - Two experiments were conducted to test the effect of nontemporal factors on duration discrimination. In Exp. 1, a forced-choice adaptive procedure with a standard duration of 400 or 800 ms was employed. It was shown that, for both auditory and visual modes, the discrimination is better with empty intervals (a silent period between two brief signals) than with filled intervals (a continuous signal), but only with shorter durations. In a second experiment, where intervals of the same duration range were employed but were presented with a single stimulus method, discrimination was better with empty than with filled intervals, and this effect applied to both ranges of duration and both sensory modes. In both experiments, discrimination was better in the auditory than in the visual mode. These data are discussed in the context of current models of timing mechanisms. PMID- 9870295 TI - Spatial information with pictures and words in visual short-term memory. AB - In a delayed matching task, the influence of spatial congruence between study and test on visual short-term memory for geometric figures and words was investigated. Subjects processed series of pictures which showed three words or three geometric figures arranged as rows or as triangular configurations. At test, the elements were presented in the identical or in the alternative configuration as at study. In the non-matching case, one of the studied elements was exchanged. The delay was 5 s. Subjects judged whether the elements were the same as during study, independent of their configuration. In Exp. 1, pictures of figures and words were mixed within one list. For both modalities, the response times were longer if the configuration at test was incongruent to the one at study. This contradicts the results of Santa, who observed effects of spatial congruency for figures, but not for words. In Exp. 2 we therefore presented the same material as in Exp. 1, but now the lists were modality-pure, as in the experiment of Santa--i.e., words and figures were shown in different lists. This time, spatial incongruency impaired recognition of the figures, but not recognition of the words. These results show that in a non-verbal context, isolated visually presented words are spatially encoded as non-verbal stimuli (figures) are. However, the word stimuli are encoded differently if the task is a pure verbal one. In the latter case, spatial information is discarded. PMID- 9870296 TI - Effects of color on perceptual and conceptual tests of implicit memory. AB - Two experiments are reported that explore why recent investigations of implicit memory failed to find any effects of color information on test performance. In the first experiment, participants studied colored pictures as well as words printed in colored ink without any memory instructions. During the test phase, a verbal and a pictorial version of a color-choice task (a conceptual priming test) were compared to two perceptual tests (word-stem completion and picture-fragment identification). Similar and significant amounts of priming to color occurred in both color-choice tasks. The perceptual tests were found to be sensitive to changes in the stimulus-presentation mode from study to test, but stimuli remaining the same color and those changed to black-and-white did not differ in the priming scores. In the second experiment, a mild division of attention was introduced in the study phase. Once again, priming to color was observed only in the verbal version of a color-choice test and not in the word-stem completion test. Dividing attention did not decrease performance on both implicit tests, whereas an explicit test of color recall for studied pictures suffered from dividing attention at encoding. It is concluded that a perceptual attribute such as color may be represented and coded by conceptual processing. Furthermore, automatic (or not attention-demanding) encoding processes may suffice for later conceptual tests of implicit memory. Previous failures to find any effects of color information on implicit performance are attributed to the use of perceptual priming tests. PMID- 9870298 TI - The Society for Adolescent Medicine: the first thirty years. Introduction. PMID- 9870297 TI - The subjective organization of input and output events in memory. AB - In order to study the organization of memory for self-performed actions, 80 participants were presented with 20 action phrases for ten consecutive study-test cycles. Enactment was manipulated both in the input phase and in the output phase by having participants say or enact the phrases during encoding and/or during testing. Enactment at input or output generally enhanced both the quantity and the accuracy of recall and also improved output monitoring. More important, subjective organization, as indexed by the tendency to recall the same two phrases successively across repeated recall tests, was significant for all conditions, even on the first pair of trials, and increased systematically with repeated study-test cycles. Enactment neither impaired nor enhanced the amount of organization, and in all conditions a positive correlation was obtained between recall and subjective organization. Some commonalities in the nature of memory organization were found across all conditions. The results suggest that enactment may lead to more differentiated memory traces, resulting in more accurate recall. Although subjective organization was clearly observed when enactment was involved, its contribution to the enhancement of recall deserves further examination. PMID- 9870299 TI - The Society for Adolescent Medicine's role in promoting education. PMID- 9870300 TI - Role of SAM in the promotion of research in adolescent health. Society for Adolescent Medicine. PMID- 9870301 TI - SAM's role in the dissemination of information. Society for Adolescent Medicine. PMID- 9870302 TI - Regionalization of SAM and evolution of the international movement in adolescent health. Society for Adolescent Medicine. PMID- 9870303 TI - History of advocacy and social policy within the Society for Adolescent Medicine. PMID- 9870304 TI - The composition of SAM: development of diversity. Society for Adolescent Medicine. PMID- 9870305 TI - Society for Adolescent Medicine: a capsule history. PMID- 9870308 TI - Determination of volatile components in fresh, frozen, and freeze-dried Padron type peppers by gas chromatography-mass spectrometry using dynamic headspace sampling and microwave desorption. AB - "Padron-type" peppers are a small variety of Capsicum annuum cultivated mainly in Galicia, Spain. To compare the effects of freezing and freeze-drying on the volatile components of Padron-type peppers, preserved samples are analyzed by means of dynamic headspace sampling on an adsorbent followed by microwave desorption into a gas chromatograph equipped with a mass spectrometric detector. Sixty-five compounds are identified, including hydrocarbons, terpenes, alcohols, phenols, ethers, aldehydes, ketones, esters, pyrroles, pyrazines, and sulfurous compounds. Fresh whole, homogenized, and freeze-dried peppers have characteristic volatile-component profiles, whereas frozen peppers have a highly variable volatile-component profile. PMID- 9870309 TI - Enantiomer analysis of chiral lactones in foods by on-line coupled reversed-phase liquid chromatography-gas chromatography. AB - A new application is proposed for the on-line coupling of reversed-phase liquid chromatography to gas chromatography (RPLC-GC) that allows the GC chirospecific analysis of gamma-lactones in fruits and commercially available fruit-containing products. The use of a programmed temperature vaporizer as an interface with the system makes the transfer of large volume fractions (i.e., 2520 microL) of aqueous eluents from LC to GC possible (speed of sample transfer, 1800 microL/min). Relative standard deviations obtained for the investigated lactones under the experimental conditions vary from 7 to 14%. The described system enlarges the LC-GC application field and overcomes the limitations reported thus far concerning the use of typical normal-phase eluents (i.e., the transfer of rather small volume fractions at low speeds of sample introduction). PMID- 9870313 TI - Interaction of human immunodeficiency virus type 1 envelope glycoprotein V3 loop with CCR5 and CD4 at the membrane of human primary macrophages. AB - We show that infection of primary monocyte-derived macrophages (MDMs) and blood lymphocytes (PBLs) by human immunodeficiency virus type 1 (HIV-1) R5 strains, but not that of PBLs by X4 strain HIV-1LAI, is inhibited by beta-chemokines RANTES and MIP-1alpha. A biotinylated disulfide-bridged peptide mimicking the complete loop of clade B consensus V3 domain of gp120 (V3Cs), but not a biotinylated V3LAI peptide or a control beta-endorphin peptide of approximately the same molecular weight (MW), was found to bind specifically to MDM membrane proteins, in particular two proteins of 42 and 62 kDa migrating as sharp bands after electroblotting onto Immobilon, and this was specifically inhibited by anti-V3 antibodies. When biotinylated V3Cs was incubated with intact MDMs, which were then washed and lysed, and the resulting material was incubated with streptavidin agarose beads and electroblotted onto Immobilon, fresh V3Cs also bound to proteins of the same molecular weight recovered in the V3Cs-interacting material. This binding was inhibited by anti-V3 antibodies, and no binding occurred with the control peptides. V3Cs also bound to soluble recombinant CD4, and CD4 monoclonal antibody Q4120 specifically recognized the V3Cs-interacting 62-kDa protein, which should thus correspond to CD4. Recombinant radiolabeled RANTES, MIP-1alpha, and MIP-1beta, but not IL-8, also bound to a 42-kDa protein on the membrane of MDMs as well as to the V3Cs-interacting 42-kDa protein, and excess unlabeled V3Cs inhibited such binding. This protein was also recognized by antibodies to CCR5, the RANTES/MIP-1alpha/MIP-1beta receptor. These data show that V3Cs binds to MDM membrane proteins that comprise CD4 and CCR5, and that multimolecular complexes involving at least gp120 V3, CD4, and CCR5 are formed on the surface of MDMs as part of V3-mediated postbinding events occurring during HIV-1 infection. PMID- 9870314 TI - Inhibition of HIV type 1 infection with a RANTES-IgG3 fusion protein. AB - The natural ligands for the chemokine receptors CCR5 (RANTES, MIP-1alpha, and MIP 1beta) and CXCR4 (SDF-1) can act as potent inhibitors of infection by the human immunodeficiency virus type 1 (HIV-1) at the level of viral entry. Unlike antibody-mediated inhibition, chemokine-mediated inhibition is broadly effective. Different HIV-1 strains can utilize the same coreceptor(s) for viral entry and, therefore, can be blocked by the same chemokine(s). HIV-1 strains that are highly resistant to neutralization by V3-specific antibodies are sensitive to inhibition by chemokines. Therefore, the use of chemokine-derived molecules constitutes a potential therapeutic approach to prevent infection by HIV-1. We have generated a fusion protein between RANTES and human IgG3 (RANTES-IgG3). The effectiveness of RANTES-IgG3 inhibition of infection by HIV-1 was similar to that of rRANTES. Inhibition of HIV-1 by RANTES-IgG3 was specific for CCR5-dependent but not CXCR4 dependent HIV-1 isolates. Fusion of a chemokine to an IgG moiety offers two desirable properties with respect to the recombinant chemokine alone. First, IgG fusion proteins have extended half-lives in vivo. Second, molecules with IgG heavy chain moieties may be able to cross the placenta and potentially induce fetal protection. PMID- 9870315 TI - A cocktail of Mycobacterium bovis BCG recombinants expressing the SIV Nef, Env, and Gag antigens induces antibody and cytotoxic responses in mice vaccinated by different mucosal routes. AB - Recombinant live Mycobacterium bovis BCG strains (rBCG) expressing different human immunodeficiency virus (HIV) or simian immunodeficiency (SIV) antigens could be good candidates for the development of vaccines against AIDS. To develop effective HIV/SIV vaccines, humoral and cellular immune responses directed against multiple antigens may be essential for the control of the infection. In this study we immunized BALB/c mice via different mucosal routes (oral, aerogenic, nasal, and rectal) with a mixture of three rBCG strains expressing, respectively, the entire SIVmac251 Nef protein, and large fragments of the Env and Gag proteins. All routes of immunization studied induced immunoglobulin A (IgA) antibodies against mycobacterial PPD, SIV Env, and SIV Gag antigens in feces and bronchial lavages as well as specific immunoglobulin G (IgG) in serum. Strong, specific cytotoxic responses of splenocytes against Nef, Env, and Gag was observed whatever the mucosal route of immunization. Therefore, mucosal vaccination with a cocktail of rBCG strains induces local, specific IgA, systemic IgG, and systemic CTLs against the three SIV antigens expressed. Rectal and oral routes seemed the most appropriate route of vaccination to be used to protect against SIV infection. PMID- 9870316 TI - Correlates of apoptosis of CD4+ and CD8+ T cells in tonsillar tissue in HIV type 1 infection. AB - The immunopathogenesis of human immunodeficiency virus type 1 (HIV-1) infection has been associated with increased death by apoptosis of T cell subsets. In the present study, we have examined correlates of apoptosis of CD4+, CD8S+CD28+, and CD8+CD28- T cells in tonsillar lymphoid tissue in persons with HIV-1. Single-cell suspensions of tonsillar lymphocytes were analyzed by flow cytometry to determine the fraction of cells showing typical characteristics of apoptosis as well as the expression of activation markers within the live and the apoptotic cell populations. The proportion of cells carrying infectious provirus was quantified by limiting dilution analysis. Compared with uninfected controls, apoptosis of both CD4+ and CD8+ T cells was enhanced in HIV-1 infection and was higher among CD8+ than among CD4+ T cells. Apoptosis of CD28-cells was more prevalent than apoptosis of CD28+ cells for both CD4+ and CD8+ T cells. Occurrence of apoptosis of CD4+ T cells correlated with provirus levels and proportional expression of the activation marker HLA-DR. Apoptosis of CD8+CD28+ cells correlated with expression of the activation markers CD69 and HLA-DR while apoptosis within CD8+CD28- cells did not correlate with any of the studied parameters. Although apoptosis was much more prevalent among CD8+ than CD4+ T cells, CD8+ T cells still accumulated in tonsillar lymphoid tissue in persons with HIV-1. Our data may be interpreted to suggest that apoptosis of CD4+, CD8+CD28+, and CD8+CD28- cells in tonsillar tissue is regulated by different mechanisms and the results are of importance to our understanding of the immunopathogenesis of HIV-1 infection. PMID- 9870317 TI - The HTLV type I sequence from an asymptomatic German blood donor is related to sequences from South America. AB - In most parts of Europe only a limited number of sporadic cases of HTLV-I infections have been identified. So far, the few cases found in Germany were individuals from endemic areas or with relations to endemic areas. Here we report an HTLV-I infection from an asymptomatic female German blood donor whose only known potential risk was a former partner from South America, where HTLV-I is known to be endemic. The DNA sequence of the LTR region was determined and a phylogenetic analysis indeed suggested homologies with HTLV-I sequences from South America. PMID- 9870318 TI - Complete sequence of an infectious molecular clone derived from a Spanish HIV type I isolate. PMID- 9870319 TI - Zidovudine therapy and HIV type 1 mutations in children with symptomatic HIV type 1 infection: effect of switching to didanosine or zidovudine plus didanosine therapy. Italian Multicenter Study Group on HIV Mutations in Children. AB - Type and prevalence of zidovudine (ZDV) resistance mutations in HIV-1-infected children in clinically stable condition and on ZDV monotherapy were analyzed to evaluate the effect of switching to didanosine (ddI) monotherapy or to ZDV plus ddI on the pattern of mutations and on the clinical outcome. Monthly clinical and laboratory controls for HIV-1 infection status were performed; at enrollment and every 4 to 6 months after treatment randomization mutant proviral sequences were evaluated in all the children, whereas viral burden was performed only in a small subgroup of patients randomly selected in each of the three treatment groups. ZDV resistance-associated proviral DNA mutations were defined as low-level resistance (LLR) mutations or medium/high-level resistance (MHLR) mutations; clinical outcome was considered as stable or deteriorating. Results showed that at entry into the study the duration of ZDV therapy was significantly correlated with the presence of mutations, and that the level of resistance given by mutations was associated with the severity both of symptoms and immunodeficiency. After randomization to treatment, in patients with mutations that confer LLR a better clinical outcome with ddI monotherapy than with ZDV plus ddI and ZDV alone was observed in the subsequent 6 months, whereas in patients with mutations that confer MHLR no significant difference among the three treatment groups was found. Data showed also that levels of viral burden at the time of changing therapy are related to clinical outcome if measured by plasma viral load. These results suggest that genotypic resistance assays, together with viral load, may prove useful for rational treatment decisions both at the start of therapy and with failure. PMID- 9870320 TI - Effect of ribavirin on zidovudine efficacy and toxicity in vitro: a concentration dependent interaction. AB - Zidovudine (ZDV) is converted to its active triphosphate (ZDVTP) by intracellular kinases. The intermediate ZDV monophosphate (ZDVMP) is believed to play a major role in ZDV toxicity. Manipulation of ZDV phosphorylation is a possible therapeutic strategy for altering the risk-benefit ratio. Here we investigate whether combining RBV with ZDV is able to modulate efficacy and toxicity of ZDV. We have measured the intracellular activation of ZDV (0.3 microM) in the absence and presence of ribavirin (RBV; 2 and 20 microM) in Molt 4 and U937 cells. MTT cytotoxicity of ZDV (10-1000 microM) was also measured with and without RBV (2 microM) in Molt 4 and U937 cells. Measurement of endogenous deoxythymidine triphosphate (dTTP) allowed investigation of the dTTP/ZDVTP ratio. The antiviral efficacy of ZDV in combination with RBV (2 microM) was assessed by HIV p24 antigen measurements. In the presence of RBV (2 and 20 microM) a decrease in total ZDV phosphates was observed, owing mainly to an effect primarily on ZDVMP rather than the active ZDVTP. RBV also increased endogenous dTTP pools in both cell types, resulting in an increase in the dTTP/ZDVTP ratio. ZDV alone significantly reduced p24 antigen production, with an IC50 of 0.34 microM. Addition of RBV increased the IC50 approximately fivefold (1.52 microM). However, at higher concentrations of ZDV (10 and 100 microM) the antagonistic effect of RBV (2 microM) on ZDV was lost. The RBV-mediated decrease in ZDVMP may explain the reduction in ZDV toxicity when combined with RBV (2 microM). Cytotoxicity of ZDV was reduced in the presence of RBV (2 microM) at all concentrations in both cell lines, probably owing to saturation of ZDVTP formation. The interaction of ZDV and RBV is concentration dependent. PMID- 9870321 TI - Role of preimmunization virus sequences in cellular immunity in HIV-infected patients during HIV type 1 MN recombinant gp160 immunization. AB - The effect of patient preimmunization virus sequences on CTL responses during gp160 immunization were studied. Ten HLA-A2+, HIV+ asymptomatic patients with CD4+ T cells >500/mm3 were given two courses of HIV-1 MN rgp160 vaccine over a 2 year period. Envelope epitope-specific CTL responses, using PBMCs, were measured against peptide-coated autologous B lymphoblastoid cell lines. Optimum CTL epitopes were determined by HLA-A2-binding affinity of 9- to 10-mer peptides containing the HLA-A2.1-binding motif. Ten of the high- or intermediate-binding peptides were conserved among >50% of reported clade B HIV strains. These peptide specific CTL activities and the patient virus sequences in peptide-coding regions were monitored. Six patients showed envelope peptide-specific CTL responses, which correlated with the presence of whole envelope antigen-specific CTL responses. Five of these patients, who showed responses to epitopes in the gp41 region (aa 814-824), had preimmunization virus similar to the vaccine sequence in this region. Three patients who did not show these epitope-specific responses had initially different sequences in the HIV gene encoding that region. The epitope specific CTL responses appear to reflect recall responses, as only patients infected with virus containing the vaccine sequence developed them and they could be recalled with a second set of vaccine injections. This appears to be reminiscent of the concept of T cell "original antigenic sin." This vaccine was also immunogenic as measured by gp160-specific lymphocyte-proliferative responses. However, increased immune responses did not impact the HIV load or CTL epitope sequences during therapy. PMID- 9870322 TI - Horizontal transmission of human immunodeficiency virus type 1 from father to child. AB - An unusual case of human immunodeficiency virus type 1 (HIV-1) infection in a child was studied. The child, identified as HIV-1 infected at 5 years of age, lived with his parents and a 3-year-old sister. HIV-1 infection was excluded in the mother and sister, but confirmed in the father, who was unaware of his infection and was in good health, apart from an atopic dermatitis on the face and limbs. A portion of the HIV-1 proviral envelope gene was amplified from the father's and child's peripheral blood cells, and the amplified products were cloned and sequenced. Phylogenetic analysis disclosed that the father's and child's viral sequences clustered together, and were clearly distinct from the sequence sets obtained from six epidemiologically unlinked mother-child HIV-1 infected pairs included in the analysis. HIV-1 variability was lower in the child's sequence set than in the father's, and the variability between father's and child's sequences was significantly lower than that found between epidemiologically unlinked cases (p < 0.001). An uncommon APGR motif on the tip of the V3 domain was found in both the father's and child's viral clones. These data, together with the epidemiological investigations, strongly suggest that the child acquired the infection from his father, possibly by exposure to bleeding skin lesions. PMID- 9870324 TI - Graffiti by any other name? PMID- 9870325 TI - The certification process in adolescent medicine. AB - The first and second certifying examinations in adolescent medicine were administered jointly by the American Board of Pediatrics (ABP) and the American Board of Internal Medicine (ABIM) on November 15, 1994, to 295 candidates, and on November 18, 1997, to 197 candidates including 170 who were taking the examination for the first time. This report describes the certification process and the characteristics of those taking the first and second examinations in adolescent medicine. The purposes of certification in adolescent medicine, consistent with the purposes of specialty boards, are to improve the quality of patient care during the second decade of life, expand and improve training to include a comprehensive and scientifically based approach to the care of young people as they enter adulthood, and stimulate faculty development and research for the problems of the adolescent population (1). Certification addresses these goals by establishing rigorous standards for training, verifying the knowledge base of candidates through a written examination, and requiring periodic recertification. This certification program recognizes a growing number of physicians with expertise in the medical problems of adolescents and the emergence in the past 30 years of a body of knowledge about the health problems, the effects of rapid biologic and psychologic change, and the interaction of biologic, psychologic, social, and environmental forces that characterize this unique developmental period (2). PMID- 9870323 TI - HIV type 1 subtypes in Malaysia, determined with serologic assays: 1992-1996. AB - We investigated the molecular epidemiology of HIV-1 subtypes in Malaysia among injecting drug users (IDUs) and sexual transmission risk groups, using serologic and genetic techniques. Frozen sera collected at a general hospital, a blood bank, several drug treatment centers, and an STD clinic in Kuala Lumpur, between 1992 and 1996, were investigated retrospectively. V3 peptide serotyping and monomeric gp120 capture serotyping were used to study 89 known HIV-1-infected subjects. The methods differentiate subtypes B, E, and C. V3 peptide and gp120 capture results were comparable. No subtype C-specific reactive sera were found; one specimen was dually reactive for subtypes C and B, using the V3 peptide ELISA; and four were durally reactive for subtypes E and C using this assay. Genotypic analysis of HIV-1 gag RNA in serum was done on a subset of subjects and confirmed serologic findings. HIV-1 subtypes differed significantly by risk category: of 53 IDUs, 29 (55%) were infected with subtype B and 19 (36%) were infected with subtype E, 3 (6%) were dually reactive, and 2 (4%) were not typable. Of 36 persons with heterosexual risks, 29 (81%) were infected with subtype E, 5 (14%) were infected with subtype B, and 2 (5%) were not typable. Persons with IDU risks were significantly more likely to be infected with subtype B than were those with sexual risks (OR 5.89; 95% CI, 1.94-18.54; p < 0.001). Subtypes B and E of HIV-1 appear to predominate in Malaysia; subtype B was more prevalent among IDUs; subtype E was more prevalent among all other groups. These results may have important HIV-1 vaccine implications. PMID- 9870326 TI - Scientific abstracts at the Society for Adolescent Medicine meetings: a 3-decade comparison. AB - PURPOSE: The purpose of this study was to review abstracts accepted for presentation at the annual national meeting of the Society for Adolescent Medicine (SAM) over the past 3 decades for subject content and research design. METHODS: Retrospective review was conducted of research abstracts for 3 years, selected at random, for the 1990s and then matched at 10-year intervals for the previous 2 decades. The major categories for subject content were: medical, psychosocial, health services, and miscellaneous; for research design categories were: retrospective, observational, experimental, and miscellaneous. Subsequent publication was also assessed. RESULTS: The most notable change over the 3 decades in subject content was a shift from medical topics (58% to 19%) to psychosocial topics [20% to 52% (p < .001)] of abstracts, the latter largely accounted for by increases in topics pertaining to high-risk behaviors. There was a substantial decrease in abstracts concerning medical aspects of chronic illness (p < .001), well-adolescent care, growth and development, and psychosocial aspects of chronic illness (p < .02). Regarding research design, the proportion of retrospective studies decreased over the 3 decades (p < .02); the most prevalent design was observational, with an increase in studies using established databases (p < .02). Less than 15% represented an experimental design. Studies using qualitative analysis appeared for the first time in the 1990s. The percentage of presentations which eventuated in published reports increased from 35% to 63% over the 3 decades (p < .02). CONCLUSION: A significant shift occurred in subject content of scientific abstracts from a medical to a psychosocial emphasis. The proportion of retrospective designs decreased, and qualitative analyses emerged. The likelihood of later publication of the studies increased. PMID- 9870327 TI - Gallagher lecture. Thirty years in pursuit of the magic bullet. PMID- 9870328 TI - Health counseling in adolescent preventive visits: effectiveness, current practices, and quality measurement. AB - Most morbidity and mortality among adolescents results from their participation in health-compromising behaviors. Recent guidelines for clinical adolescent preventive services recommend that primary care clinicians routinely screen for and counsel adolescents about these behaviors, identify and address related social, psychological, and biologic factors. Office-based counseling can influence adult health behaviors, but little is known about the effectiveness of office-based counseling for adolescents. In this review we: (a) evaluate available information about the effectiveness of office-based health counseling to improve outcomes; (b) report what is known about the health counseling adolescents receive from primary care clinicians; and (c) critically review different approaches that have been, or might be, used to measure the content and quality of health counseling provided during adolescent medical visits. With the emphasis on accountability in the current health care environment, evidence supporting the effectiveness of counseling is needed to justify investment in this aspect of clinical adolescent preventive services. Challenges to studying the effectiveness of health counseling include the lack of well-defined theory based models for adolescent office-based counseling, the complexity of measuring counseling quality, and the many factors that influence adolescent outcomes. Reliable and valid measures of counseling quality are needed both to study and to ensure the quality of counseling received by adolescents. No single measure can be expected to fully capture counseling quality, although patient self-report deserves further development. PMID- 9870329 TI - Review of options for primary prevention of eating disturbances among adolescents. PMID- 9870330 TI - Adolescent males involved in pregnancy: associations of forced sexual contact and risk behaviors. AB - PURPOSE: To determine whether sexually active adolescent males who report being the victim of forced sexual contact and engaging in health risk and problem behaviors are more likely to report getting someone pregnant. METHODS: In 1995, 4159 students in Grades 9-12 in 59 randomly selected public high schools in Massachusetts were anonymously surveyed using the Youth Risk Behavior Survey (YRBS). Data were analyzed for 824 sexually active males. Demographic variables and indicators of sexual behavior, pregnancy, violence, and suicide were assessed. Data were analyzed with multiple logistic regression. RESULTS: A total of 12.0% of sexually active males reported having been involved in a pregnancy. The proportion of males who reported getting a partner pregnant increased with age. Of the sample, 8.1% gave a history of having had sexual contact against their will. Of those who reported forced sexual contact, 36.4% reported having been involved in a pregnancy; of the males who did not report a history of forced sexual contact, 9.4% were involved in a pregnancy (CV = 0.23; p < 0.00001). Based on multiple logistic regression, forced sexual contact [odds ratio (OR) 3.56; 95% confidence interval (CI) 1.79-7.09], frequency of weapon carrying on school property (OR 1.39; 95% CI 1.18-1.64), number of cigarettes smoked per day (OR 1.22; 95% CI 1.08-1.38), number of sexual partners in the previous 3 months (OR 1.43; 95% CI 1.25-1.65), and condom nonuse at last intercourse (OR 1.80; 95% CI 1.06-3.02) correctly classified 89.9% of the males who were involved in a pregnancy. CONCLUSION: This study highlights the association between health-risk and problem behaviors, forced sexual contact, and involvement in pregnancy among sexually active male high school students. In our analysis, a history of forced sexual contact was associated with a higher risk of high school males' involvement in pregnancy. These results strongly suggest the importance of screening sexually active males for a history of forced sexual intercourse and health risk and problem behaviors in the effort to prevent teenage pregnancy and childbearing. PMID- 9870331 TI - Pregnancy among three national samples of runaway and homeless youth. AB - PURPOSE: To compare estimates of the prevalence of pregnancy among runaway and homeless youth between the ages of 14 and 17 years in various settings with each other and with youth in the general population. METHODS: Comparisons used three surveys of youth: (a) the first nationally representative survey of runaway and homeless youth residing in federally and nonfederally funded shelters, (b) a multicity survey of street youth, and (c) a nationally representative household survey of youth with and without recent runaway and homeless experiences. RESULTS: Youth living on the streets had the highest lifetime rates of pregnancy (48%), followed by youth residing in shelters (33%) and household youth (<10%). CONCLUSIONS: Shelter and street youth were at much greater risk of having ever been pregnant than were youth in households, regardless of whether they had recent runaway or homeless experiences. Such youth need comprehensive services, including pregnancy prevention, family planning, and prenatal and parenting services. PMID- 9870333 TI - The weight-conscious adolescent: body image, food intake, and weight-related behavior. AB - PURPOSE: To explore how adolescents who try to lose weight differ from those who do not in relation to body image, food intake, knowledge about food, and sources of information about food, nutrition, and weight loss. METHODS: Data were collected from 12-15-year-old school students in North Queensland, Australia by questionnaire. RESULTS: Boys who attempted weight loss reduced sweet foods and snacks, while concurrently increasing healthy foods such as fruit, yogurt, and low-fat milk. Girls who attempted weight loss ate breakfast, lunch, snacks, milk, bread, meat, and many sweet and fatty foods less often than other girls. Some reduction in meals, snacks, core foods, and sweet fatty foods, but not savory fatty foods, remained after the weight loss attempt. Boys attempting weight loss had better knowledge about high-fat and high-sugar foods than other boys, but there was no difference for girls. Girls attempting weight loss were more likely than other girls to think that core foods such as bread, potatoes, and dairy products should be reduced, whereas boys were more likely to think that sweet foods should be reduced. The majority of students who had attempted weight loss during the previous year considered themselves overweight, while few weight conscious students were satisfied with their bodies. CONCLUSIONS: The weight conscious adolescents in this study, especially the girls, exhibited restrictive eating practices and a preoccupation with a slim image. Adolescents need a food culture based on foods to eat rather than foods to avoid, and an understanding of suitable weight-control measures. PMID- 9870334 TI - Cannabinoid modulation of wide dynamic range neurons in the lumbar dorsal horn of the rat by spinally administered WIN55,212-2. AB - The effects of spinally administered cannabinoids on nociceptive responses of wide dynamic range (WDR) neurons in the lumbar spinal cord were investigated in urethane-anesthetized rats. Noxious thermal stimulation was applied with a Peltier device to regions of the ipsilateral hindpaw corresponding to the receptive fields of isolated neurons. WIN55,212-2 (100 microg, i.t.), applied topically on the dorsal spinal surface, suppressed noxious heat-evoked activity in spinal WDR neurons. By contrast, responsiveness was unchanged following administration of either vehicle or WIN55,212-3, the receptor-inactive enantiomer. WIN55,212-2, administered intrathecally to separate rats, produced antinociceptive effects in the tail-flick test with a time course and efficacy that paralleled the suppression of noxious heat-evoked activity. These results suggest that cannabinoid modulation of spinal nociceptive processing involves direct actions in the spinal dorsal horn and is related to the antinociceptive effects of intrathecally administered cannabinoids. PMID- 9870332 TI - Substance use and risky sexual behavior among homeless and runaway youth. AB - PURPOSE: To (a) characterize human immunodeficiency virus (HIV)-related risk behaviors of homeless youth; (b) determine whether substance use is associated with risky sexual behavior in this population; and, if so, (c) explore explanations for this relationship. METHODS: A purposive sample of 327 homeless youth (ages 14-21 years) in Washington, DC, were surveyed in 1995 and 1996. Survey items were adapted from items used in a national study of adult substance use and sexual behavior and measured global (lifetime) and event-specific (most recent sexual encounter) behaviors. RESULTS: Sexual activity with many partners, "survival" sex, and substance use were common. However, needle use was rare, and consistent condom use was evident in half the sample. Nearly all correlations between global measures of substance use and risky sex were statistically significant, but only a few of the event-specific correlations were significant. Marijuana use during the most recent sexual encounter was associated with nonuse of condoms, but this relationship disappeared in the multivariate model. However, crack use during the last encounter was associated with condom use; this relationship remained significant in the multivariate model. Lack of motivation to use condoms, longer histories of sexual activity and homelessness, symptoms of drug dependency, not discussing HIV risks with partner, and being female were also associated with nonuse of condoms. CONCLUSIONS: Homeless youth do use condoms, even within the context of substance use and casual sex. Results suggest that prevention and targeted intervention efforts have had some positive effect on this population, but young homeless women are in need of targeted prevention. Finally, additional research is needed to investigate the observed relationship between crack use and condom use in this sample. PMID- 9870335 TI - Inhibition of prostanoid synthesis protects against neuronal damage induced by focal ischemia in rat brain. AB - Changes in prostanoids concentration and effects of the non-specific COX inhibitor indomethacin on prostanoids levels and extension of tissue damage were studied following focal ischemia induction in the fronto-parietal region of rat brain. Ischemia was induced in animals bearing a transcerebral microdialysis probe by injection of Rose Bengal, a photosensitive dye, followed by light activation. Prostanoid levels were determined in the dialysate using immunoenzymatic techniques. PGD2 levels rose significantly up to 237+/-22 pg/ml compared to a basal level measured before ischemia induction which was below the detection limit. TXB2 changes were smaller and had a different time course. Treatment with indomethacin abolished the ischemia-induced PGD2 release and reduced the extent of injury to the area by 43+/-3.7%. These results suggest that prostanoid release may play an important role in neurodegenerative processes and that cyclooxygenase inhibitors may contribute to protect against cerebral tissue damage. PMID- 9870336 TI - Immunocytochemical evidence for a distinct GFAP-negative subpopulation of astrocytes in the adult rat hippocampus. AB - In order to establish the relative distribution of a GFAP-negative population of astrocytes, and its change in gliotic tissue, sections of the stratum radiatum of the CA1 hippocampal layer of male, adult, Wistar rats were analyzed by immunocytochemical methods. Ten micrometer-thick sections were triple-stained to detect nuclei, glial fibrillary acidic protein (GFAP) and glutamine synthetase (GS). In another set of experiments, the rats received a one-time intraperitoneal injection of kainic acid that caused epileptic seizures. With the use of a behavioral protocol, animals with substantial neuronal loss in the pyramidal layer were selected. Five days after the injection these rats were analyzed similarly to control rats. We find that GFAP-positive cells are a subpopulation of GS-positive cells and that the GFAP-negative subpopulation is quite large (40%). After gliosis the density of GFAP-negative, GS-positive cells stays stable, whereas the GFAP-positive population triples. These experiments confirm electrophysiological experiments showing a distinct, GFAP-negative subset of astrocytes that remains consistent even after injury-induced gliosis and accompanying up-regulation of GFAP. PMID- 9870337 TI - Concurrent activation of hippocampal glycine and polyamine sites of the N-methyl D-aspartate receptor synergistically reverses working memory deficits in rats. AB - Intrahippocampal administration of the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (0.18 microg/side) significantly increased the number of errors in the working memory task with a three-panel runway setup. The increase in working memory errors by intrahippocampal MK-801 was significantly attenuated by concurrent infusion of D-cycloserine (1.0 microg/side) or spermidine (10 microg/side), agonists of the glycine and polyamine modulatory sites on the NMDA receptor/channel complex, respectively. Combined injection of the behaviorally ineffective doses of 0.1 microg/side D-cycloserine and 0.32 microg/side spermidine synergistically reduced intrahippocampal MK-801-induced increase in working memory errors. The combination of D-cycloserine and spermidine also synergistically attenuated the increase in working memory errors resulting from intrahippocampal injection of the muscarinic acetylcholine receptor antagonist scopolamine (3.2 microg/side). These results suggest that positive modulation of the NMDA receptor/channel through activation of the glycine and polyamine sites can synergistically compensate deficiency of hippocampal NMDA and muscarinic receptor-mediated neurotransmission involved in working memory function. PMID- 9870338 TI - Effect of infusion of vasoactive intestinal peptide (VIP)-antisense oligodeoxynucleotide into the third cerebral ventricle above the hypothalamic suprachiasmatic nucleus on the hyperglycemia caused by intracranial injection of 2-deoxy-D-glucose in rats. AB - We examined the effect of the infusion of a vasoactive intestinal peptide (VIP) antisense oligodeoxynucleotide into the third cerebral ventricle above the hypothalamic suprachiasmatic nucleus (SCN) using osmotic minipump for 3 days (0.2 nmol/ml per h) on the hyperglycemic response to intracerebroventricular injection of 2-deoxy-D-glucose (2DG) (80 micromol) in rats. After the infusion of the VIP antisense the inhibition of VIP expression in the SCN was observed in association with suppressions of the hyperglycemia, hyperglucagonemia and relative hypoinsulinemia due to the 2DG injection. Furthermore, additional intracranial injection of VIP (4 nmol) restored these responses to the 2DG injection in rats treated with the VIP antisense. These findings suggest that VIP neurons in the SCN are involved in the regulation of glucose metabolism. PMID- 9870339 TI - Evaluation of anesthetic effects on parameters for the in situ rat brain perfusion technique. AB - Studies of drug distribution to brain should be controlled for the experimental method used. Numerous methods have been employed to ascertain brain distribution and many of these approaches use anesthetic agents. The in situ rat brain perfusion method is one of the most sensitive and widely used methods for evaluating brain distribution profiles. There has been no evaluation of the effects of anesthetic agents on parameters associated with this method (i.e. cerebral perfusion fluid flow, brain vascular volume and blood-brain barrier permeability). We evaluated the effects of the anesthetic agents pentobarbital and ketamine combinations on these baseline parameters. The results suggest that the anesthetic agent has no effect on these parameters and anesthetic selection is open to the choice of the investigator when using the perfusion method. PMID- 9870340 TI - Different systems in human brain are involved in presemantic discrimination of pictures as revealed by event-related potentials. AB - It is uncertain if the several brain areas in response to presemantic picture processing are functionally homogeneous. Subjects were asked to discriminate visual stimulus pairs and event-related potentials (ERPs) were recorded from the scalp. A line-drawing picture (S1) was followed by another stimulus (S2) 600 ms later. S2 was the same as S1 in condition 1, a reversed picture of S1 in condition 2, the corresponding Chinese word in condition 3 and the syllabic alphabet of S1 in condition 4. Amplitudes of ERPs over right posterior scalp with time window from 102-180 ms after the onset of S2 in condition 1 and 2 were significantly larger than condition 3 and 4. The N2 component on the right anterolateral scalp with time window from 182-240 ms was significantly larger in condition 1 than other conditions, whereas the N2 on the left anterolateral scalp with time window from 262-350 ms in condition 2, 3 and 4 was significantly larger than condition 1. It is suggested that pictures and words are processed in different systems at the presemantic stage: repetitive and physically deviant stimuli evoke different responses in different brain regions. PMID- 9870341 TI - Localization of nitric oxide-synthesizing neurons sending projections to the dorsal raphe nucleus of the rat. AB - The origin of the nerve fibers immunoreactive for neuronal nitric oxide synthase (nNOS) in the rat dorsal raphe nucleus (DRN) was determined by combining the use of cholera toxin subunit b (CTb) as a retrograde tracer and nNOS immunohistochemistry with a monoclonal anti-nNOS antibody. Double labeled CTb nNOS cell bodies were distributed from the rostral diencephalon to the caudal medulla oblongata, in about 20 areas of the brain. Several of the areas displaying double labeled cells are known for their involvement in the control of the sleep-wake cycle and/or transmission of nociception. PMID- 9870342 TI - Both medial prefrontal and amygdala central nucleus lesions abolish heart rate classical conditioning, but only prefrontal lesions impair reversal of eyeblink differential conditioning. AB - Rabbits with lesions of either medial prefrontal cortex (mPFC) or amygdala central nucleus (ACN) were compared with sham-lesioned animals during differential and reversal classical conditioning of the eyeblink (EB) and heart rate (HR) response. Lesions of the mPFC, but not ACN, produced a severe impairment in EB reversal conditioning, but neither lesion affected original discrimination. However, both mPFC and ACN lesions produced a severe attenuation of accompanying HR decelerations during both initial differentiation and reversal. These results suggest that mPFC processing of Pavlovian conditioning contingencies affects not only the autonomic component of learning but preservative somatomotor conditioning as well, whereas ACN processing affects only the autonomic component. PMID- 9870343 TI - Brief exposure of mice to 60 Hz magnetic fields reduces the analgesic effects of the neuroactive steroid, 3alpha-hydroxy-4-pregnen-20-one. AB - Relatively weak, extremely low frequency (ELF), magnetic fields have been shown to exert a variety of biological effects, although the modes of action remain to be established. Neuroactive steroids and neurosteroids have been shown to produce a diverse range of rapid centrally mediated behavioral and physiological effects that are reported to be sensitive to magnetic fields. Here we show that brief exposure of male mice to an ELF magnetic field (30 min, 60 Hz, 141 microT peak) significantly reduces the analgesic effects arising from intracerebroventricular (i.c.v.) administration of the centrally produced allylic neuroactive steroid, 3alpha-hydroxy-4-pregnen-20-one (3alphaHP) and that the dihydropyridine (DHP) calcium channel antagonists, diltiazem and nifedipine, block the inhibitory effects of the 60 Hz ELF on 3alphaHP-induced analgesia. These results indicate that exposure to 60 Hz ELF affects the analgesic effects of neuroactive steroids such as 3alphaHP through alterations in calcium channel function. These findings raise the possibility that ELF magnetic fields may, in part, exert their actions through effects on diverse neuroactive steroid modulated processes. PMID- 9870344 TI - The non-competitive N-methyl-D-aspartate-antagonist memantine does not affect segmental mono- and polysynaptic reflexes in man. AB - Studies in rats have shown that the polysynaptic flexor reflex (FR) but not the monosynaptic reflexes are affected by N-methyl-D-aspartate (NMDA) receptor antagonists. Theoretically, the suppression of FR might be caused by an alteration of the spinal nociceptive neurons. To investigate, whether the non competitive NMDA receptor antagonist memantine interferes with nociception in man, we studied both its effect on pain perception and on FR. In a double-blind study 14 male subjects were randomly assigned to either placebo or memantine (30 mg p.o.) treatment. H-reflex (HR) and FR as well as pain and tolerance threshold were determined prior to and 6 h after drug intake. Contrary to expectations, there were no differential treatment effects either on FR threshold and magnitude or on pain and tolerance thresholds or the HR amplitude. PMID- 9870346 TI - Up-regulation of glial cell line-derived neurotrophic factor (GDNF) expression in regenerating muscle fibers in neuromuscular diseases. AB - Glial cell line-derived neurotrophic factor (GDNF) has been shown to exert a target-derived trophic factor for motor neurons. Immunohistochemical analyses revealed that expression of GDNF in regeneration muscle fibers was up-regulated in polymyositis (PM) and Duchenne type muscular dystrophy (DMD). Reverse transcriptase polymerase chain reaction (RT-PCR) analyses showed that the full length GDNF was up-regulated in PM and DMD muscle; normal muscle exhibited mostly truncated GDNF. The results indicate that the GDNF expression is regulated in regeneration of human skeletal muscle. PMID- 9870345 TI - D-2-amino-5-phosphonovalerate, and NMDA receptor antagonist, blocks induction of associative long-term potentiation of the flexion reflex in spinal cat. AB - The effect of the NMDA receptor antagonist D-2-amino-5-phosphonovalerate (APV) on classical conditioning of the flexion reflex in spinal cat was examined. Animals perfused intrathecally with artificial cerebral spinal fluid (CSF) containing APV exhibited flexion reflex potentiation during the conditioning period that was not significantly different from cats receiving artificial CSF alone. However, the APV group exhibited no signs of reflex potentiation during the 2.5 h retention period, in contrast to the CSF alone group. The results suggest that NMDA receptor activation plays a critical role in the induction of associative long term potentiation of flexion reflexes in spinal cat. PMID- 9870347 TI - Insulin-like growth factor system in serum and cerebrospinal fluid in patients with multiple sclerosis. AB - The insulin-like growth factor (IGF) system influences oligodendrocyte survival, myelination, and immune functions. We examined whether alterations in the circulating IGF system occur in multiple sclerosis (MS), a chronic inflammatory demyelinating disease of the central nervous system. We measured concentrations of IGF-I, IGF-II, and insulin-like growth factor binding proteins -1, -2, and -3 in both serum and cerebrospinal fluid from MS patients and age- and sex-matched controls. IGFBP-1 was not detectable in cerebrospinal fluid. We found no significant differences in any of the other components between patients with MS and controls. PMID- 9870348 TI - 1998: the centenary of the discovery of the Golgi apparatus. AB - 1998 is the year of the centenary of the discovery of the Golgi apparatus. This event is considered in its historical context: the first cell theory of 1838 1839, the first polemics in cytology and the research on the cell organelles at the turn of the century. The first approaches to clarify the physiological significance of the apparatus is traced from Golgi (1909) to Bowen (1929). PMID- 9870349 TI - A general approach to desalting oligosaccharides released from glycoproteins. AB - Desalting of sugar samples is essential for the success of many techniques of carbohydrate analysis such as mass spectrometry, capillary electrophoresis, anion exchange chromatography, enzyme degradation and chemical derivatization. All desalting methods which are currently used have limitations: for example, mixed bed ion-exchange columns risk the loss of charged sugars, precipitation of salt by a non-aqueous solvent can result in co-precipitation of oligosaccharides, and gel chromatography uses highly crosslinked packings in which separation of small oligosaccharides is difficult to achieve. We demonstrate that graphitized carbon as a solid phase extraction cartridge can be used for the purification of oligosaccharides (or their derivatives) from solutions containing one or more of the following contaminants: salts (including salts of hydroxide, acetate, phosphate), monosaccharides, detergents (sodium dodecyl sulfate and Triton X 100), protein (including enzymes) and reagents for the release of oligosaccharides from glycoconjugates (such as hydrazine and sodium borohydride). There is complete recovery of the oligosaccharides from the adsorbent which can also be used to fractionate acidic and neutral glycans. Specific applications such as clean-up of N-linked oligosaccharides after removal by PNGase F and hydrazine, desalting of O-linked glycans after removal by alkali, on-line desalting of HPAEC-separated oligosaccharides and beta-eliminated alditols prior to electrospray mass spectrometry, and purification of oligosaccharides from urine are described. PMID- 9870351 TI - Hydrolase-catalyzed beta-mannosylations. AB - Transmannosylations catalysed by beta-mannosidase from snail viscera or beta galactosidase from Aspergillus oryzae were accomplished with 4-nitrophenyl beta-D mannopyranoside as donor substrate. With suitable hydrophobic acceptor molecules preferentially beta1-4-linked disaccharides were obtained. The activities of both glycosidases in buffer cosolvent mixtures were determined, and conditions for their immobilization were elaborated and optimized. A model of the enzymic transfer mechanism is suggested. PMID- 9870350 TI - Deglycosylation by gaseous hydrogen fluoride of mucus glycoproteins immobilized on nylon membranes and in microtiter wells. AB - Strongly reacting antibodies specific for defined mucin gene products are often directed against the mucin protein backbone of the heavily glycosylated serine/threonine rich regions. A prerequisite for the use of such antibodies is often the complete removal of the oligosaccharides from the protein. This paper describes an efficient one-step deglycosylation method using gaseous hydrogen fluoride on nylon blotting membranes and microtiter wells. PMID- 9870352 TI - Effect of cysteine modifications on the activity of the 'small' Clostridium perfringens sialidase. AB - The 'small' (43 kDa) sialidase of Clostridium perfringens is inhibited by low concentrations of mercury ions. For the investigation of possible functional roles of the enzyme's four cysteine residues at the amino acid positions 2, 282, 333 and 349, they were separately altered to serine by site-directed mutagenesis. The four mutant sialidases expressed in E. coli and purified by metal chelate chromatography were markedly reduced in specific activity when compared to the wild-type enzyme but with the exception of C282S exhibited similar K(M)-values indicating an unchanged mode of substrate binding. The substrate specificity was also conserved for C2S, C282S, and C333S. Only the C349S sialidase exhibited a higher relative activity with colominic acid and the alpha2,6-linked sialic acid of sialyllactose compared to the alpha2,3-linked isomer than the other mutants. Chemical modifications with the thiol-blocking reagents N-ethylmaleimide (NEM), p chloromercuribenzoate (pCMB) and HgCl2 had little effect on the C282S sialidase, e.g., 6% inhibition by 5 mM NEM compared to reductions in activity between 65 and 90% for the wild-type and other mutant enzymes, supporting the idea that among the enzyme's cysteines, Cys-282 has the highest structural or functional significance. The results also explain the higher mercury tolerance of Salmonella typhimurium and Clostridium tertium sialidases, which have the positions equivalent to Cys-282 altered to Val and Thr, respectively, indicating that the thiol group of Cys-282, despite being situated near the active site, is not involved in catalysis. PMID- 9870353 TI - Effect of tunicamycin on the activity and immunoreactivity of ascorbate oxidase (Cucurbita pepo medullosa) expressed in cultured green zucchini cells. AB - Ascorbate oxidase activity and immunoreactivity were evaluated in crude tissue extracts obtained from callus cell cultures induced by green zucchini sarcocarp and grown in the presence of tunicamycin, a powerful N-glycosylation inhibitor. Tunicamycin at 2 or 4 microg ml(-1) blocked cell growth within a couple of weeks, although a sustained cell viability was observed in the same period. A significant inhibition of total protein synthesis was observed at 10 and 15 days of culture time, with a decrease of 30% and 43% respectively when cells were grown in the presence of 2 microg ml(-1) tunicamycin, and of 48% and 57% respectively when the tunicamycin concentration was 4 microg ml(-1). After the same culture times ascorbate oxidase specific activity assayed in crude tissue extracts showed increases of about 1.9-fold and 3.5-fold (10 days) and 1.7-fold and 3.1-fold (15 days) at 2 and 4 microg ml(-1) tunicamycin, respectively. Ascorbate oxidase mRNA levels, however, did not appreciably differ between control and treated samples, measured at the same growing times. Lectin-blot, based on the use of concanavalin A, indicated a marked decrease of glycosylated proteins in tunicamycin-treated cultures. As judged by immunoblot, anti-native ascorbate oxidase antibodies scarcely recognized the enzyme expressed in tunicamycin-treated cells; on the contrary, anti-deglycosylated ascorbate oxidase antibodies were more reactive to the enzyme expressed in tunicamycin-treated cultures. PMID- 9870354 TI - Purification and characterization of GDP-L-Fuc: N-acetyl beta-D-glucosaminide alpha1-->6fucosyltransferase from human blood platelets. AB - c-6-L-Fucosyltransferase (alpha1,6FucT; EC 2.4.1.68) from human platelets, the enzyme that is released into serum during coagulation of blood, was purified 100,000-fold. The purification required three sequential chromatographic steps: chromatofocusing, affinity column chromatography on GnGn-Gp(asialo-aglacto transferrin glycopeptide)-CH-Sepharose, and gel filtration of Sephadex G-200. The final preparation contained a protein that migrated as a single discrete band Mr of 58,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under non-reducing conditions, and as a single enzymatically active peak Mr of 58,000 in gel filtration. Although the purified enzyme utilized the biantennary GnGn-Gp as substrate, it was twice as active with the triantennary oligosaccharide when the Man alpha1,3 antenna was substituted with GlcNacbeta1,4. On the other hand the tetraantennary oligosaccharide was not a preferred substrate. The Km values for the substrate asialo-agalactotransferrin glycopeptide, and GDP-L-fucose were 29 and 28 microM, respectively. The optimum pH of the enzyme was 6.0. The activity of alpha1,6FucT was abolished in the presence of beta-mercaptoethanol. Divalent cations such as Mg2+ and Ca2+ activated, but Cu2+, Zn2+ and Ni2+ strongly inhibited the activity. PMID- 9870355 TI - The ruminant parasite Haemonchus contortus expresses an alpha1,3 fucosyltransferase capable of synthesizing the Lewis x and sialyl Lewis x antigens. AB - Glycoproteins from the ruminant helminthic parasite Haemonchus contortus react with Lotus tetragonolobus agglutinin and Wisteria floribunda agglutinin, which are plant lectins that recognize alpha1,3-fucosylated GlcNAc and terminal beta GalNAc residues, respectively. However, parasite glycoconjugates are not reactive with Ricinus communis agglutinin, which binds to terminal beta-Gal, and the glycoconjugates lack the Lewis x (Le(x)) antigen or other related fucose containing antigens, such as sialylated Le(x), Le(a), Le(b) Le(y), or H-type 1. Direct assays of parasite extracts demonstrate the presence of an alpha1,3 fucosyltransferase (alpha1,3FT) and beta1,4-N-acetylgalactosaminyltransferase (beta1,4GalNAcT), but not beta1,4-galactosyltransferase. The H. contortus alpha1,3FT can fucosylate GlcNAc residues in both lacto-N-neotetraose (LNnT) Galalpha1-->4GlcNAcbeta1-->3Galbeta1-->4Glc to form lacto-N-fucopentaose III Galbeta1-->4[Fuca1-->3]GlcNAc beta1-->3Galbeta1-4GIc, which contains the Le(x) antigen, and the acceptor lacdiNAc (LDN) GalNAcbeta1-->4GlcNAc to form GalNAc beta1-->4[Fualpha1-->3]GlcNAc. The alpha1,3FT activity towards LNnT is dependent on time, protein, and GDP-Fuc concentration with a Km 50 microM and a Vmax of 10.8 nmol-mg(-1) h(-1). The enzyme is unusually resistant to inhibition by the sulfhydryl-modifying reagent N-ethylmaleimide. The alpha1,3FT acts best with type 2 glycan acceptors (Galbeta1-->4GlcNAcbeta1-R) and can use both sialylated and non-sialylated acceptors. Thus, although in vitro the H. contortus alpha1,3FT can synthesize the Le(x) antigen, in vivo the enzyme may instead participate in synthesis of fucosylated LDN or related structures, as found in other helminths. PMID- 9870356 TI - The aberrant expression of Lewis a antigen in intestinal metaplastic cells of gastric mucosa is caused by augmentation of Lewis enzyme expression. AB - Immunohistochemical staining showed an aberrant expression of Le(a) antigen in the intestinal metaplastic glands of the gastric mucosa of secretors, as reported by others. In this study, we have demonstrated for the first time that the Lewis enzyme is well colocalized with Le(a) antigen, indicating that the Lewis enzyme is responsible for Le(a) antigen synthesis in the gastric mucosa. The staining intensity of the Lewis enzyme was much stronger in the cells with intestinal metaplasia than the cells without metaplasia, regardless of the secretor status. The amount of transcript of the Lewis gene was related to the degree of metaplasia; i.e., the more severe the metaplastic change was, the more abundantly the transcripts of the Lewis gene were expressed. This augmentation of the Lewis enzyme in metaplastic tissues was also confirmed by Western blotting analysis using a specific antibody against the Lewis enzyme. We conclude that intestinal metaplastic change of gastric mucosa is usually accompanied by a marked augmentation of the Lewis enzyme expression, which results in the enhanced expression of Le(a) antigens, particularly in secretors. PMID- 9870357 TI - Isolation of the receptor for Amaranthus leucocarpus lectin from murine peritoneal macrophages. AB - The receptor for Amaranthus leucocarpus lectin from CD-1 resident macrophages was purified with affinity chromatography with biotin labeled A. leucocarpus lectin and using avidin-agarose as affinity matrix. The receptor is a glycoprotein of 70 kDa that contains 18% of sugar by weight; it is mainly composed of galactose and N-acetyl-D-galactosamine in its saccharidic portion, and lacks sialic acid; the protein is rich in glycine, serine and alanine and lacks cysteine residues. The amino terminus of the receptor is blocked. By ionic strength chromatography on a mono P column in anionic form we purified three isoforms from the affinity purified receptor, each showing quantitative differences in glycosylation. The A. leucocarpus lectin receptor is identified only in resting, not activated, macrophages suggesting that it plays a role in activation mechanisms of macrophages. PMID- 9870358 TI - Characterization of oligosaccharides from an antigenic mannan of Saccharomyces cerevisiae. AB - Mannans of the yeast Saccharomyces cerevisiae have been implicated as containing the allergens to which bakers and brewers are sensitive and also the antigen recognized by patients with Crohn's disease. A fraction of S. cerevisiae mannan, Sc500, having high affinity for antibodies in Crohn's patients has been characterized by NMR spectroscopy followed by fragmentation using alkaline elimination, partial acid hydrolysis and acetolysis. The released oligosaccharides were separated by gel filtration on a Biogel P4 column and analyzed by fluorescence labeling, HPLC and methylation analysis. The relationship between structure and antigen activity was measured by competitive ELISA. The antigenic activity of the original high molecular weight mannan could be ascribed to terminal Manalpha1-->3Manalpha1-->2 sequences which are rarely found in human glycoproteins but were over-represented in Sc500 compared to other yeast mannans. PMID- 9870359 TI - Different O-glycosylation of respiratory mucin glycopeptides from a patient with cystic fibrosis. AB - The O-linked oligosaccharides from three fractions of highly glycosylated mucin glycopeptides obtained from sputum of a patient with cystic fibrosis were characterized and compared regarding size, composition, sequence and when possible linkage positions. Neutral and sialic acid-containing glycans were permethylated and analyzed by high-temperature GC-MS and MALDI-MS, showing more than 60 different oligosaccharides with a size of up to 15 monosaccharide units. Some of the observed oligosaccharides are novel for respiratory secretions, one being a trifucosylated heptasaccharide with the proposed structure: Fuc-Gal-4(Fuc 3)GlcNAc-(Fuc-)Gal-3GalNAcol. The glycosylation of two of the glycopeptide fractions was similar with regard to the neutral and sialylated oligosaccharides despite their different origins from the sol or gel phase. Analysis of the sulfated oligosaccharides by FAB-MS/MS indicated that the gel fraction contained C-6 linked sulfate groups while the two sol fractions also contained C-3 linked sulfate. The results suggest the presence of different glycosylated mucin domains, probably originating from different mucin glycoforms and/or apoproteins in the airway of cystic fibrosis patients. PMID- 9870361 TI - The efficiency of vertical transmission of Neospora caninum in dairy cattle analysed by serological techniques. AB - The association of Neospora caninum infections with cattle families was examined in a dairy cattle herd with sporadic abortions using three different serological tests. Cattle seropositive for N. caninum clustered in six families, three of which encountered abortions. In absence of age-related differences in the N. caninum seroprevalence, the family association of N. caninum infection indicated that congenital infection represented the predominant route of transmission in this herd. Fourteen (93%) out of 15 descendants of 10 seropositive cows were seropositive themselves. Only one female calf of a seropositive cow remained seronegative and gave birth to a calf which was tested seronegative again. Only one seronegative cow that had two seronegative descendants also gave birth to one seropositive calf. This was the only indication for potential postnatal transmission that occurred in the herd. The results of this study suggest that the N. caninum-infection can be maintained over several generations at a nearly constant prevalence level, apparently without a need for dispersion by an definitive host. PMID- 9870360 TI - Glycation changes the charge distribution of type I collagen fibrils. AB - In aging and diabetes, glycation of collagen molecules leads to the formation of cross-links that could alter the surface charge on collagen fibrils, and hence affect the properties and correct functioning of a number of tissues. The electron-optical stain phosphotungstic acid (PTA) binds to positively charged amino acid side-chains and leads to the characteristic banding pattern of collagen seen in the electron microscope; any change in the charge on these side chains brought about by glycation will affect the uptake of PTA. We found that, upon glycation, a decrease in stain uptake was observed at up to five regions along the collagen D-period; the greatest decrease in stain uptake was apparent at the c1 band. This reduction in PTA uptake indicates that the binding of fructose leads to an alteration in the surface charge at several sites along the D-period. Not all lysine and arginine residues are involved; there appear to be specific residues that suffer a loss of positive charge. PMID- 9870362 TI - Haemoparasite infections in newly introduced dairy cattle in Loei Province, Thailand: Trypanosoma evansi antigen levels by ELISA referring to abortion. AB - An antigen-detection enzyme-linked immunosorbent assay (Ag-ELISA) based on affinity-purified polyclonal antibody was utilised as an immunodiagnosis for Trypanosoma evansi infections in cattle. Five hundred pregnant heifers were introduced in Loei province, Thailand and a total of 271 samples were collected at 37 farms during four visits over a year commencing a month after the introduction. Each time the haematocrit centrifuge technique (HCT) was carried out as a field diagnosis for T. evansi, and sera were examined for trypanosomal antigen levels by Ag-ELISA. At the first sampling over 80% of the cattle were positive for trypanosome antigens by Ag-ELISA although the titres were low. Soon after, aborted cases at the late stage of pregnancy were reported and at the second sampling in the rainy season, 25.5% of the cattle sampled were found to harbour T. evansi by HCT. This time the infection rate by Ag-ELISA was 52.9% with high antigen levels. Between the first and second sampling nine cattle out of 51 aborted, which was suspected to be due to T. evansi. As soon as treatment with a trypanocidal drug was started, abortion cases decreased. However, the infection rate remained high during the rainy season when Tabanus flies were active. As the climate became cool and dry, the antigen levels in the area lowered and the positive rate by Ag-ELISA dropped to 32.3%. PMID- 9870363 TI - PCR primer evaluation for the detection of trypanosome DNA in naturally infected goats. AB - The buffy coat of 76 roaming goats from the Bansang and Missira regions in Gambia, was examined for the presence of trypanosomes. From these animals, extractions from dry blood samples on filter paper were subjected to PCR using three different primer sets, ORPHON5J, GOL and TVW, specific for Trypanosoma brucei/Trypanosoma evansi, Trypanosoma congolense and Trypanosoma vivax, respectively. PCR results for T. congolense were 100% concordant with buffy coat examination. Besides the three T. vivax buffy coat-positive samples, another 15 yielded positive with the TVW primers. The ORPHON5J primers yielded no positive results. Analyses with the GOL primers of putatively negative samples, yielded aberrant band patterns whose diagnostic significance still remains to be determined. PMID- 9870364 TI - Alterations of serum lipid, lipoprotein and inflammatory cytokine profiles of rabbits infected with Trypanosoma brucei brucei. AB - In the study to analyze the pathophysiology of chronic salivarian trypanosomosis, the infection of rabbits with Trypanosoma brucei brucei produced a three-phase alteration of serum lipid and lipoprotein profiles. In the earliest stages, during the first 2 weeks post-infection (p.i.), serum triglycerides and very low density lipoproteins (VLDL) began to increase, high density lipoproteins (HDL) began to decrease, while a more rapid increase was observed in low density lipoproteins (LDL). Serum phospholipids increased moderately at 2 weeks p.i. preceded by a transient decrease at 1 week p.i. In the second phase, from 2 weeks p.i., the infected animals developed a condition consisting of a gradual accumulation of VLDL together with high triglycerides, high LDL and low HDL. Serum cholesterols increased moderately from 2 weeks p.i. The third phase was a convalescent stage between 7 and 10 weeks p.i. The alterations of triglyceride, VLDL and LDL levels suggested that suppression of lipolytic enzymes in charge of hydrolysis of serum triglycerides was initiated by 2 weeks p.i., and then became seriously impaired corresponding with the progression of disease. Serum tumor necrosis factor alpha (TNFalpha) was continuously detected from 1 week p.i., and for at least 5 additional weeks in all the infected animals. Serum interleukin (IL)-1-like activity was occasionally detected, and IL-6-like activity increased in the middle stages of infection in some of the infected animals. The derangement of lipid metabolism appeared to be associated with the period of TNFalpha induction following infection. PMID- 9870365 TI - Stage-specific activity in vitro on the Theileria infection process of serum from calves treated prophylactically with buparvaquone. AB - An in vitro method for testing activity of buparvaquone in serum on the infection and development of Theileria in its bovine host mononuclear cells is described and results compared with the effect exhibited in vivo. Serum samples were collected over a time course from calves in a clinical trial of 5 mg kg(-1) buparvaquone prophylaxis on Theileria annulata or T. parva experimental infection. To evaluate drug levels and persistence in each animal for a period of 14 days and its effect on the early infection stages, the sera were tested on established macroschizont infected cell lines and against the in vitro infection and development process of the sporozoite and trophozoite stages of the two Theileria species. Results from the in vitro assays show that buparvaquone in serum can completely prevent the establishment of Theileria infection during the first 48 h after administration at 5 mg kg(-1). After seven days, levels are sufficient to delay the establishment of infection. The drug is more effective in the prevention of the de novo development of the parasite in cells than against established macroschizont infected cell culture. At low concentrations, it is more effective against T. parva than against T. annulata. Drug effect peaks during the first 24 h but residual effect persists for 14 days, particularly against T. parva infection. These novel findings demonstrate how high doses of buparvaquone could over-protect calves if used in the 'infection-and-treatment' method of immunisation when drug is administered prophylactically at the same time as infection with live sporozoites. It is suggested that in certain high Theileria risk situations there may be potential for the immunoprophylactic use of buparvaquone without simultaneous infection. The in vitro assay itself has been shown to be of value as a model for Theileria establishment in cattle. PMID- 9870367 TI - Enzyme-linked immunosorbent assay (ELISA) for detection of anti-Trypanosoma evansi equine antibodies. AB - The standardization of ELISA for the detection of anti-Trypanosoma evansi antibodies in naturally and experimentally infected horses is described. Bayesian analysis was used to establish the cutoff between positive and negative sera. In order to determine the assessment of the ELISA test, the results obtained were compared with those from an IFA. A relative sensibility of 98.39%, a specificity of 95.12% and a predictive value of 96.83% were determined. The standardized technique was used to evaluate the antibody production against trypanosome in an experimentally infected equine, in which the sera converted 15 days after infection. The test was also used for a study of sera prevalence in a non-random sample from two different populations. A prevalence of 81.7% in workhorse and 57.14% in stable horses was found. PMID- 9870366 TI - Immunohistochemical detection of parasite antigens in Theileria parva-infected bovine lymphocytes. AB - Theileria parva is the causal agent of East Coast fever (ECF), a fatal disease of cattle characterized by pyrexia, transient lymphadenopathy and panleukopenia. We have evaluated monoclonal antibodies (mAb) against three distinct antigens (p67, PIM and p32) of the parasite as immunohistological reagents for monitoring the kinetics of infection in cattle. Bovine lymphocytes were stained with the mAb at various intervals after infection in vitro and in vivo. The p67 sporozoite surface antigen was detected in only a small percentage of both, in vitro and in vivo infected cells. In contrast, expression of the polymorphic immunodominant molecule (PIM) of the parasite proved a useful indicator of infection and staining was correlated with the results of Giemsa analysis. PIM was detected from day 3 in in vitro-infected cells, but was not detected until day 5 in vivo after challenge with a 70% lethal dose of stabilized sporozoite. The p32 antigen was expressed only late in infection in vivo and its expression was associated with the development of merozoites. Less than 20% of in vitro-infected cells expressed p32. The immunohistochemical staining with anti-PIM mAb was found to be a useful tool for analysis of T. parva infection kinetics in cattle. PMID- 9870368 TI - Glucose uptake in Oesophagostomum dentatum and the effect of oxfendazole. AB - The uptake of 14C-glucose by adult Oesophagostomum dentatum was characterised. The uptake was a non-linear function of external glucose concentration. The maximum velocity of uptake (Vmax) was 0.964 nmol/100 mg dry weight (dw)/5 min, and the transport constant (Kt) was 10.02 microM. When phlorizin, phloretin and 3 O-methylglucose were tested for their effects on the uptake of 14C-glucose, phloretin and 3-O-methylglucose produced significant inhibitions, indicating that the uptake was mediated and occurred by facilitated diffusion. Exposure of the worms to oxfendazole prior to incubation with 14C-glucose did not affect the uptake of glucose. In another experiment worms were incubated with unlabelled glucose and the external glucose concentration was measured enzymatically. During a 7 h incubation period, the quantity of glucose remaining in the incubation medium of oxfendazole exposed worms was significantly greater than in the control group. It was concluded that oxfendazole did not influence the process of 14C glucose uptake, but might induce changes in the parasite leading to a reduced ability to deplete the incubation medium of glucose. PMID- 9870369 TI - The effect of previous cold storage on the subsequent recovery of infective third stage nematode larvae from sheep faeces. AB - An investigation undertaken to determine the effect of previous cold storage on the recovery of third stage larvae of gastrointestinal nematodes of sheep, showed that increasing periods of exposure of faeces to 4 degrees C resulted in decreasing numbers of larvae subsequently recovered from them. Differences in the abilities of the eggs of the various genera, to survive such treatment, were found to lead to significant changes in the percentage generic compositions of their third stage larvae--in some cases following the prior refrigeration of faecal samples for as little as 24 h. These results suggest that where larval cultures are intended to provide estimates of the proportions of the various worm eggs in the faeces of sheep harboring mixed gastrointestinal nematode burdens, they should be performed only on freshly collected samples. PMID- 9870371 TI - Capillary electrophoresis in drug analysis. AB - Capillary electrophoresis has become one of the advanced analytical methods for drugs in pharmaceutical, therapeutic, diagnostic and forensic applications. This review discusses key issues and provides key references to the topic of drug analysis using capillary electrophoresis. It gives readers a brief summary of the current status of the technology and serves as an editorial for the paper symposium "Capillary electrophoresis in drug analysis". PMID- 9870370 TI - Prevalence of IgG and IgM antibodies specific to Toxoplasma gondii in cats. AB - IgG and IgM antibodies specific to Toxoplasma gondii were determined simultaneously by IFAT in cats. Group 1 included 357 cats with various diagnoses, but negative for FeLV and FIV. Specific IgG antibodies in titres ranging from 10 to 2560 (mean 247) were detected in 61.3% animals, specific IgM antibodies were found only in one cat (0.28%) in titre 1:40 showing an IgG titre of 160. Oocyst shedding was demonstrated in one cat (0.28%) showing an IgG titre of 1:40. Group 2 included 33 cats positive for FeLV or FIV. IgG antibodies in titres ranging from 10 to 640 (mean 101) were found in 63.6% cats. Neither IgM antibodies nor shedding of T. gondii oocysts were detectable. Clinically unapparent toxoplasmosis, manifested by the presence of IgG antibodies is frequent in cats; the opportunistic character of the infection is important. PMID- 9870372 TI - Strategies for capillary electrophoresis: method development and validation for pharmaceutical and biological applications. AB - This review is in support of the development of selective, reproducible and validated capillary electrophoretis (CE) methods. Focusing on pharmaceutical and biological applications, the successful use of CE is demonstrated by more than 800 references, mainly from 1994 until 1998. Approximately 80 recent reviews have been catalogued. These articles sum up the existing strategies for method development in CE, especially in the search for generally accepted concepts, but also looking for new, promising reagents and ideas. General strategies for method development were derived not only with regard to selectivity and efficiency, but also with regard to precision, short analysis time, limit of detection, sample pretreatment requirements and validation. Standard buffer recipes, surfactants used in micellar electrokinetic capillary chromatography (MEKC), chiral selectors, useful buffer additives, polymeric separation media, electroosmotic flow (EOF) modifiers, dynamic and permanent coatings, actions to deal with complex matrices and aspects of validation are collected in 20 tables. Detailed schemes for the development of MEKC methods and chiral separations, for optimizing separation efficiency, means of troubleshooting, and other important information for key decisions during method development are given in 19 diagrams. Method development for peptide and protein separations, possibilities to influence the EOF and how to stabilize it, as well as indirect detection are considered in special sections. PMID- 9870373 TI - Analyte identification in capillary electrophoretic separation techniques. AB - A review on applications of on-line hyphenation in capillary electrophoresis and capillary electrochromatography for the identification of migrating analytes is presented. There is an urgent need for unambiguous analyte identification by combining spectral information and observed migration times, because the parameters influencing the migration times and separation efficiencies in these separation techniques are not easily controlled, especially when real samples containing unknown interferences have to be analyzed. The spectrometric techniques covered here are ultraviolet and visible radiation (UV/Vis) absorption, fluorescence including fluorescence line-narrowing spectroscopy, Raman spectroscopy, nuclear magnetic resonance and mass spectrometry. Attention is essentially confined to literature reports in which the extra information provided by the detector is really used for identification purposes, especially in real-life samples, while the interfacing as such and analyte detectabilities in standard solutions are only briefly discussed. This article covers an extensive fraction of the literature published on this topic until the beginning of 1998. PMID- 9870374 TI - Drug analysis by capillary electrophoresis and laser-induced fluorescence. AB - This review briefly presents the different laser-induced fluorescence detectors, outlines the different dyes used to derivatize molecules which are used with capillary electrophoresis/laser-induced fluorescence (CE-LIF), and provides an overview and current status of CE-LIF in drug analysis. PMID- 9870375 TI - A literature assessment of sample pretreatments and limits of detection for capillary electrophoresis of drugs in biological fluids and practical investigation with some antimalarials in plasma. AB - A literature survey on published reports of the determination of drugs in biological fluids shows that all methods of sample pretreatment have been used and that the limits of detection achieved vary widely, ranging from low ngcm(-3) to microgcm(-3). The most widely used injection method was hydrodynamic and, in the majority of cases, whenever low detection limits were achieved, this was a result of preconcentration during the sample pretreatment. Only a small proportion of the reported methods employed electrokinetic injection and utilised the field amplified sample injection (FASI) techniques. An experimental investigation of the alternative hydrodynamic and electrokinetic injection methods for a small set of antimalarial drugs is reported. It was found that electrokinetic injection with FASI from an acetonitrile-water matrix produced dramatic improvements in detection limits. This improvement could not, however, be achieved when the drugs were in plasma using protein precipitation, liquid liquid extraction or solid phase extraction pretreatment methods. This highlights the importance of sample pretreatment in utilising the potential sensitivity of capillary electrophoresis with electrokinetic injection. PMID- 9870376 TI - Separation of chiral compounds by capillary electrophoresis. AB - This review presents the different chiral selectors used in capillary electrophoresis (CE) for the separation of enantiomers. The use of charged cyclodextrins, crown ethers, polysaccharides, proteins, natural and synthetic micelles, macrocyclic antibiotics and ergot alkaloids is discussed in detail. Neutral native and derivatized cyclodextrins are not treated because several review articles have already been published on this topic. Recent developments like the application of two chiral selectors in the same background electrolyte are highlighted. PMID- 9870377 TI - Method development strategies for the enantioseparation of drugs by capillary electrophoresis using cyclodextrins as chiral additives. AB - General strategies for the development of capillary electrophoretic methods for the enantiomeric separation of basic, acidic or neutral drugs were developed. For all kinds of compounds, the use of a buffer made of 100 mM phosphoric acid adjusted to pH 3 with triethanolamine and containing anionic and/or uncharged cyclodextrin (CD) derivatives as chiral selectors was recommended. Two different optimization schemes depending on the acidic or basic character of the analytes, were elaborated. For most basic compounds present in cationic form at pH 3, enantiomeric separation could be achieved in the normal polarity mode. Different beta-cyclodextrin derivatives were first tested at a given concentration. Five derivatives were found to be particularly useful for enantioseparations in capillary electrophoresis (CE): the anionic carboxymethyl-beta-CD (CMCD) and sulfobutyl-beta-CD (SBCD) and the neutral dimethyl-beta-CD (DMCD), trimethyl-beta CD (TMCD) and hydroxypropyl-beta-CD (HPCD). After selection of the most suitable CD, its concentration was optimized with respect to chiral resolution. If necessary, a further improvement in resolution could often be obtained for the enantiomers of cationic solutes by increasing the buffer pH from 3 to 5 using CMCD as chiral additive. Another possible alternative for enhancement in chiral resolution was the addition of metharlol or cyclohexanol to the buffer. For acidic drugs, essentially present in uncharged form at pH 3, and for neutral solutes, anionic CD derivatives such as SBCD or CMCD were first tested at a given concentration in the reversed polarity mode. Dual systems, based on the simultaneous addition of a charged CD (SBCD or CMCD) and a neutral CD (TMCD or DMCD), could then be investigated for resolution improvement. After optimization of the CD concentrations, the use of dual systems with CMCD at pH 5 could also be tested if necessary, especially for very weak acidic and neutral drugs. By applying these optimization strategies, 48 of the 50 drugs examined as model compounds could be fully enantioseparated by CE in short analysis times (usually less than 10 min). PMID- 9870378 TI - Assessment of pharmacodynamic and pharmacokinetic characteristics of drugs using microdialysis sampling and capillary electrophoresis. AB - Microdialysis sampling combined with capillary electrophoresis is emerging as a new approach in drug studies. It allows the continuous monitoring, in vivo or in vitro, of changes in free endogenous compounds as well as in drug substances, following the administration of pharmacological agents. The low volume requirement of capillary electrophoresis for injection allows the collection of dialysates during short sampling times, leading to a precise temporal description of drug-induced biochemical changes or pharmacokinetics. Various protocols can be used for analyzing endogenous compounds and drug substances in microdialysis samples. Capillary electrophoresis with laser-induced fluorescence detection often affords the high sensitivity level which is needed in most studies. Furthermore, the direct on-line coupling of microdialysis, derivatization of samples, and electrophoretic analysis now brings a separation-based biosensor, allowing a real-time description of chemical events with a high molecular specificity. Microdialysis sampling combined with capillary electrophoresis has recently been used to assess pharmacodynamic and pharmacokinetic characteristics of various drugs in animal studies; it may also represent a new approach in clinical pharmacology in the near future. PMID- 9870379 TI - Capillary electrophoresis for therapeutic drug monitoring. AB - Therapeutic drug monitoring is commonly used in both the ambulatory and hospital patient care settings. Routine measurement of concentrations of therapeutic agents in biological fluids is critical for certain drugs to maintain therapeutic benefit with minimizing drug-associated toxicities. Many analytical laboratory techniques are currently available to measure drug concentrations in biological samples. Recently there has been an increased interest in the use of capillary electrophoresis (CE) for measuring concentrations of therapeutic drugs in patient samples. However, while there are numerous reports of CE being used to measure drug concentrations in solution and pharmaceutical dosage forms, there are relatively few reports of the use of CE for measuring therapeutic agents in patient samples. The purpose of this paper is to provide an overview of methods currently used to measure therapeutic drugs in patient samples along with possible future trends for the use of CE in therapeutic drug monitoring. PMID- 9870380 TI - Capillary electrophoresis for therapeutic drug monitoring of antiepileptics. AB - We examined the use of capillary electrophoresis for therapeutic drug monitoring of antiepileptic drugs. Micellar electrokinetic capillary chromatography (MEKC) with a diode array detector simultaneously determined concentrations of zonisamide, a new type of antiepileptic drug, and phenobarbital, phenytoin and carbamazepine, typical antiepileptic drugs, in human serum. Zonisamide levels in human serum obtained by MEKC correlated well with levels obtained by high performance liquid chromatography. The serum levels of phenobarbital, phenytoin and carbamazepine determined by MEKC were almost equal to those obtained by fluorescence polarization immunoassay. The reproducibility of separation and quantification with MEKC for intra- and inter-day assays were appropriate. This MEKC method could provide a simple and efficient therapeutic drug monitoring method for antiepileptic drugs, especially in patients treated with a combination of zonisamide and other antiepileptic drugs. MEKC may be an attractive method for therapeutic drug monitoring, because of its specificity of separation, automation of procedure, ease of method development, low cost, small aqueous buffer amounts, speed of analysis, small injection volume and high environment-directed performance. PMID- 9870381 TI - Use of capillary electrophoresis methods to characterize the pharmacokinetics of antisense drugs. AB - As antisense drugs become mature for clinical trials, analytical techniques to analyze antisense DNA in biological media for characterization of their pharmacokinetics will be in demand. Due to the superior resolving power of capillary gel electrophoresis (CGE), CGE will likely be a preferred method in quantifying intact oligonucleotides as well as the putative metabolic products. Nonetheless, biological mediums can influence the stability of the gel column, making a CGE assay time-consuming. In one approach, high-performance liquid chromatography (HPLC) was used to quantify the total amount of antisense compounds to increase the sample throughput and CGE was used to determine the relative percentage of the intact and metabolic species on specific samples. Alternatively, extensive sample pretreatment procedures were performed and the samples were quantified and characterized directly by CGE alone with the use of an internal standard. Both methods have been used to characterize the pharmacokinetics of antisense compounds. This review focuses on the instrumental and technical aspects of analyzing antisense DNA in biological mediums using CGE either as a single or a combined method towards better understanding of the pharmacokinetics of antisense DNA. Moreover, the newer analytical technologies of capillary electrophoresis (CE), which hold great potential to be used for pharmacokinetic applications, such as the replenishable sieving matrix combined with an innovative coupling approach and microchip CE, will also be explored. PMID- 9870382 TI - A critical evaluation of the application of capillary electrophoresis to the detection and determination of 1,4-benzodiazepine tranquilizers in formulations and body materials. AB - Studies of the capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MEKC) behaviour of 1,4-benzodiazepines have seen application in subject areas such as the development of pharmaceuticals, therapeutic drug monitoring and forensic toxicology. In the development of pharmaceuticals, pKa determinations by CZE can be used in preclinical studies whereas analytical data on the detection and determination of 1,4-benzodiazepines is of value primarily in raw material/formulation assay and in the analysis of body fluids in clinical studies. The capillary electrophoresis (CE) techniques, which generally have inferior limits of detection (LOD) to rival techniques such as gas chromatography (GC) and high-performance liquid chromatography (HPLC), are particularly applicable in forensic toxicology where reasonably high concentrations of these drugs can be encountered. It is anticipated that, with the interfacing of CZE and capillary electrochromatography (CEC) with mass spectrometry (MS) techniques, the excellent selectivity of CZE and particularly CEC will be effectively combined with the sensitivity of MS and the identification capabilities of tandem mass spectrometry (MS/MS) and MS hyphenated (MSn) techniques. PMID- 9870383 TI - Use of cyclodextrins in capillary electrophoresis: resolution of tramadol enantiomers. AB - Capillary zone electrophoresis was successfully applied to the enantiomeric resolution of racemic tramadol. Both uncoated and polyacrylamide-coated capillaries were tested for method optimization using either negatively charged or native cyclodextrins (CD) added to the background electrolyte (BGE). The resolution was strongly influenced by the CD type and concentration as well as by the pH and the concentration of the BGE. Among the CDs tested, carboxymethylated beta-cyclodextrin allowed the baseline separation of tramadol enantiomers. After the method was optimized, it was validated in a coated capillary for enantiomeric analysis of tramadol enantiomers in pharmaceutical formulation, including specificity and elution order, linearity, accuracy and precision, determination of limit of detection (LOD) and quantification (LOQ), enantiomeric purity linearity, freedom from interference, and stability of sample solutions. Precision at the target concentration was less than 2%, with an accuracy higher than 99%. Furthermore, the method was able to detect 0.3% and to quantify 1% of the minor enantiomer in the presence of the major one at the target value. PMID- 9870384 TI - Analysis of cefadroxil by micellar electrokinetic capillary chromatography: development and validation. AB - A method is developed and validated for analysis of the antibiotic cefadroxil using micellar electrokinetic capillary chromatography. It permits cefadroxil to be completely separated from ten of its known related substances within 15 min (including the washing procedure). The separation is performed in an acetate buffer (50 mM, pH 5.25) containing sodium dodecyl sulfate (SDS; 110 mM). The fused-silica capillary was 44 cm long (36 cm effective length), 50 microm ID; the voltage, 18 kV; temperature, 15 degrees C; and the detection wavelength; 254 nm. The influence of the type of buffer, buffer pH and concentration, and of the SDS concentration was investigated. The robustness of the method was examined by means of a full-fraction factorial design. The parameters for validation such as linearity, precision, limit of detection and limit of quantitation are also reported. PMID- 9870385 TI - Characterization of partitioning behavior of cephalosporins using microemulsion and micellar electrokinetic chromatography. AB - Electrokinetic chromatography (EKC) was introduced to determine the partitioning behavior of various cephalosporins (cefpim, cefpirom, cefaloridin, cefaclor, cephalexin, cefuroxim, cefotaxim) in microemulsions (ME) and micellar (MC) systems. The partitioning behavior of cephalosporins in microemulsions was characterized calculating the capacity factor. The required parameters for the determination of the capacity factor (micro(aq) and micro-me are the electrophoretic mobilities of the solutes in the aqueous phase and the microemulsion phase, micro(eff) is the effective mobility in the microemulsion solution) were measured by EKC using cationic and anionic microemulsion systems consisting of the surfactants/n-heptane/1-butanol/10 mM phosphate buffer solution, pH 7.0. Electrokinetic chromatography was shown to be a useful method to quantify the partitioning behavior of drugs in oil/water microemulsion. The logarithm of the capacity factor was correlated with the logarithm of the 1 octanol/water partitioning coefficients. PMID- 9870386 TI - Selectivity manipulation using nonaqueous capillary electrophoresis. Application to tropane alkaloids and amphetamine derivatives. AB - Non-aqueous capillary electrophoresis was investigated for its potential in the analysis of pharmaceutical compounds, namely tropane alkaloids and amphetamine derivatives. The separation of these drugs was compared in aqueous and organic media such as methanol and/or acetonitrile. Selectivity, migration times and efficiency were critically affected by the composition of the methanol/acetonitrile mixture, as well as by the nature and the concentration of the electrolyte. In particular, the migration orders of two positional isomers, littorine and hyoscyamine, were inverted in the presence of trifluoroacetic acid in the nonaqueous medium. The same behavior was observed for amphetamine methamphetamine and for two methylenedioxyamphetamine derivatives. PMID- 9870387 TI - Two examples of rapid and simple drug analysis in pharmaceutical formulations using capillary electrophoresis: naphazoline, dexamethasone and benzalkonium in nose drops and nystatin in an oily suspension. AB - Capillary electrophoresis is a versatile tool in pharmaceutical analysis. In the course of a revision of the "Standardrezepturen", a German formula of standard dispensings for preparation in pharmacies, this technique has been applied to drug analysis in pharmaceutical formulations. The present paper deals with two different examples. First, naphazoline, dexamethasone and the preservative benzalkonium are quantified in nose drops without any sample preparation. Second, the antifungal antibiotic nystatin is quantified using nonaqueous capillary electrophoresis in methanol after sample preparation from an oily suspension. PMID- 9870388 TI - Analysis of vitamin formulations by electrokinetic chromatography utilizing tetradecylammonium ions as the pseudostationary phase. AB - A recently proposed method for the separation of fat-soluble vitamins by electrokinetic chromatography was further developed and investigated in the present study. The separation medium consisted of acetonitrile-water (80:20 v/v) and contained 80 mM tetradecylammonium bromide (TDA+); the content of acetonitrile served to maintain the hydrophobic vitamins dissolved during electrophoresis, while the TDA+ ions served as the pseudostationary phase. With the cathode placed at the outlet of the capillary, the fat-soluble vitamins were separated based on different hydrophobic interactions to the TDA+ ions and migrated in order of decreasing hydrophobicity prior to the electroosmotic flow. Migration time stability was significantly enhanced by the addition of 4 mM borate to the separation medium. The separation system was validated for the determination of vitamin E acetate in commercial tablets; quantitative results deviated by less than 3.5% from specified values, varying by less than 2.5% relative standard deviation (RSD) for within-day experiments, and by less than 6.5% RSD during between-day experiments. The separation system was compatible with injection solvents ranging in polarity from water to tetrahydrofuran, and was even capable of separating the water-soluble vitamins B1, B2, B12, and nicotinamide. PMID- 9870389 TI - Effects of various anionic chiral selectors on the capillary electrophoresis separation of chiral phenethylamines and achiral neutral impurities present in illicit methamphetamine. AB - In this study, various anionic chiral selectors were investigated for the capillary electrophoresis (CE) separation of six chiral phenethylamines and three achiral neutral impurities which are commonly identified in illicit methamphetamine. Analyses were carried out at pH 8 (high osmotic flow) with untreated capillaries using 25 mM chiral surfactant or 10 mM charged cyclodextrin. The chiral selectors included the micelle (R)-N dodecoxycarbonylvaline (EnantioSelect (R)-Val-1) (ES) and the cyclodextrins sulfobutyl(IV)-ether-beta-cyclodextrin (SBE(IV)-beta-CD) (BSB4), SBE(VII)-beta-CD (BSB7), SBE(XII)-beta-CD (BSB 12), SBE(IV)-gamma-CD (GSB-4), SBE(VII)-gamma-CD (GSB-7), sulfated(XI)-alpha-cyclodextrin (SU(XI)-alpha-CD (AS11), SU(VII)-beta-CD (BS7), SU(XII)-beta-CD (BS12) and SU(XIII)-beta-CD (GS13). Enantiomeric and achiral selectivity strongly depends on the size of the CD, the average degree of substitution, and the type of substitution. ES exhibits good performance for the neutral solutes, but exhibits enantiomeric selectivity only for the alpha hydroxyphenethylamines. GS13 provides the best overall enantiomeric selectivity. All fifteen solutes related to methamphetamine are simultaneously separated using BSB7. PMID- 9870390 TI - Capillary electrophoresis separation of the new anti-AIDS agents 9-(2 phosphonylmethoxyethyl)adenine and 9-(2-phosphonylmethoxyethyl)-2,6-diaminopurine in mixtures with some monoribonucleotides or the most common deoxynucleotides. AB - The present work describes an electrophoretic method for the separation and determination of the new antivirals, 9-(2-phosphonylmethoxyethyl)adenine (PMEA) and 9-(2-phosphonylmethoxyethyl)-2,9-diaminopurine (PMEDAP) in model mixtures with some monoribonucleotide isomers (3'-AMP, 2'-CMP, 3'-CMP, 3'-GMP, 2'-GMP, 3' UMP, 5'-GMP, and 5'-UMP) or with the most common deoxynucleotides (dCMP, dCDP, dCTP, dTMP, dTDP, dTTP, dGMP, dGDP, dGTP, dAMP, dADP, dATP). A fused-silica capillary tube, 75 microm ID, 67.8 cm total length (60.3 cm length to the detector), with detection at 210 nm was employed. A hydrodynamic injection for 10 s (1.5 psi vacuum) was utilized to introduce the sample, and 30 kV voltage was applied for the separation. The complete separation of PMEA and PMEDAP from the mononucleotide isomers and deoxynucleotide mixtures is possible in less that 10 min and 25 min, respectively, using 20 mM borate buffer, pH 9.9, with the addition of 10 mM beta-cyclodextrin. Efficiencies of more than 120 000 and resolution higher than 1.9 were reached for each of the compounds studied. This capillary electrophoretic procedure opens the possibility for future determination of PMEA and PMEDAP in cell pool samples. PMID- 9870392 TI - Analysis of an oligonucleotide N3'-->P5' phosphoramidate/phosphorothioate chimera with capillary gel electrophoresis. AB - N3'-->P5' phosphoramidate/phosphorothioate chimeric oligonucleotides (ODNs) are presently under investigation as potential antisense drugs. Within the field of antisense research, "second generation" chimeric ODNs have exhibited improved characteristics relative to oligonucleotides with uniformly modified backbones. The ODN of interest for this study consisted of a chemically synthesized 18-mer of mixed nucleotide base sequence with a backbone consisting of eight central phosphorothioate linkages flanked by four N3'-->P5' phosphoramidate (amidate) linkages on the 5'-end and five amidate linkages ont he 3'-end. This chimera presents analytical challenges due to the central phosphorothioate region. Here, we present a capillary gel electrophoresis (CGE) method for the analysis of the above N3'-->P5' phosphoramidate/phosphorothioate chimera. CGE was used to analyze the product prior to its purification by reversed phase - high performance liquid chromatography (RP-HPLC), and each fraction collected from the purification was similarly analyzed. An internal standard was utilized to determine the relative mobility of our product, and polyacrylamide gel electrophoresis (PAGE) analysis was used to verify CGE results. PMID- 9870391 TI - Quality evaluation by capillary electrophoresis of amphotericin B injection after filtration through various membrane filters. AB - The determination of amphotericin B, an antifungal agent, was developed using micellar electrokinetic capillary chromatography (MEKC) with a diode array detector. Repeatability and intermediate precision of MEKC analysis were acceptable. A high correlation was found between amphotericin B levels in pharmaceutical solutions obtained by MEKC and those by high-performance liquid chromatography (HPLC) (r = 0.994). This MEKC method is therefore useful for the determination of amphotericin B. The concentration of amphotericin B did not significantly change after filtration through polyethersulfone (PES, 0.2 microm) and polyvinylidene difluoride (PVDF, 0.45 microm) membrane filters. When the Fungizone injection was filtered through PES (0.2 microm) and added to 5% dextrose for injection (500 mL), particulate matters larger than 10 microm decreased by 64% to a level under the standard defined by United States Pharmacopoeia (USP XXIII). PVDF filtration (0.45 microm) did not have this effect. Our results suggest that filtration of Fungizone injection through PES (0.2 microm) membrane filters is recommended for the preparation of intravenous amphotericin B fluid. PMID- 9870393 TI - Therapeutic drug monitoring of doxorubicin in paediatric oncology using capillary electrophoresis. AB - A method for the determination of doxorubicin and its main metabolite doxorubicinol in human plasma is described. Two different sample preparation procedures are applied depending on the expected concentration: To monitor the peak plasma levels, 10 microL of plasma are deproteinated with acetonitrile. After centrifugation, the supernatant is directly applied to the capillary by hydrodynamic injection. For the determination of lower amounts of doxorubicin and its main metabolite doxorubicinol 100 microL of plasma is extracted by liquid /liquid extraction with chloroform. After evaporation of the organic phase, the sample is reconstituted in acetonitrile/water (95/5 v/v) and injected into the capillary by electrokinetic injection. Idarubicin serves as the internal standard. Laser-induced fluorescence detection with an Ar-ion laser emitting at 488 nm and a 520 nm cut-off filter is used for detection. The accuracy of the method was calculated to be 3.0% at higher concentrations and 15.0% at the limit of quantification. Reproducibility data are in accordance to the generally accepted criteria for bioanalytical methods. The limit of quantification is 2 microg/L, enabling us to monitor doxorubicin plasma levels for several days after application. Noninvasive blood sampling (from the fingertip) using heparinized capillaries was found to be a simple and convenient procedure and provides reproducible data. Initial results show high interindividual variability in doxorubicin peak plasma levels. PMID- 9870394 TI - On-line dialysis solid-phase extraction coupled to capillary electrophoresis. AB - A fully automated dialysis solid-phase extraction (SPE) sample preparation procedure is coupled on-line to capillary electrophoresis (CE) for the first time. The system is used to determine sulfonamides in serum and urine. The dialysis unit serves to remove proteins and particulate matter. Reconcentration of the analytes is performed with a small SPE column while (in)organic salts and other interferences are removed simultaneously. Finally, the analytes are desorbed and injected, via a homemade interface, into the CE system. Limits of detection (LOD) of 0.05-0.1 and 0.05-0.3 microg/mL are obtained in urine and serum, respectively. The within-day and between-day precisions are in the range of 2-6% and 3-8%, respectively, for a concentration of five times the LOD. The dialysis SPE-CE system was used over a period of six months for the analysis of over 500 serum and urine samples without problems such as clogging of the CE capillary or SPE column. PMID- 9870395 TI - Comparison of capillary electrophoresis-based immunoassay with fluorescence polarization immunoassay for the immunodetermination of methamphetamine using various methamphetamine antibodies. AB - An accurate and simple immunoassay using capillary electrophoresis (CE) with laser-induced fluorescence (LIF) was performed for the detection of methamphetamine (MA) in urine. The CE-LIF was conducted with an untreated fused silica column using antiserum and a tracer of fluorescein isothiocyanate (FITC) labeled MA. This CE-LIF system was compared with fluorescence polarization immunoassay (FPIA) in a TDx analyzer in the photo-check mode using the same FITC labeled tracer and the same antiserum. Various antibodies, not only those prepared by our own immunogens but also those from commercial sources, were screened and characterized in both assay systems with regard to sensitivity, precision, and cross-reactivity. Both systems satisfied analytical precision and gave similar cross-reactivity patterns. However, the CE-LIF-based immunoassay was approximately one order superior to FPIA in sensitivity, requiring less volume of sample, antiserum, and tracer for the assay. Considering that the FPIA system is well known to be a useful tool for screening antibodies and detecting drugs, the CE-LIF-based immunoassay system, which is seemingly more advantageous than the FPIA system, appears to have great power for the characterization of antibodies and for the detection of MA in urine. PMID- 9870396 TI - Screening for urinary amphetamine and analogs by capillary electrophoretic immunoassays and confirmation by capillary electrophoresis with on-column multiwavelength absorbance detection. AB - This paper characterizes competitive binding, electrokinetic capillary-based immunoassays for screening of urinary amphetamine (A) and analogs using reagents which were commercialized for a fluorescence polarization immunoassay (FPIA). After incubation of 25 microL urine with the reactants, a small aliquot of the mixture is applied onto a fused-silica capillary and unbound fluorescein-labeled tracer compounds are monitored by capillary electrophoresis with on-column laser induced fluorescence detection. Configurations in presence and absence of micelles were investigated and found to be capable of recognizing urinary D-(+) amphetamine at concentrations > about 80 ng/mL. Similar responses were obtained for racemic methamphetamine (MA) and 3,4-methylenedioxymethamphetamine (MDMA). The electrokinetic immunoassay data suggest that the FPIA reagent kit includes two immunoassay systems (two antibodies and two tracer molecules), one that recognizes MA and MDMA, and one that is geared towards monitoring of A. For confirmation analysis of urinary amphetamines and ephedrines, capillary electrophoresis in a pH 9.2 buffer and multiwavelength UV detection was employed. The suitability of the electrokinetic methods for screening and confirmation is demonstrated via analysis of patient and external quality control urines. PMID- 9870397 TI - Analysis of fluorescein isothiocyanate derivatized amphetamine and analogs in human urine by capillary electrophoresis in chip-based and fused-silica capillary instrumentation. AB - Amines can easily be derivatized with fluorescein isothiocyanate isomer I (FITC) and analyzed by capillary electrophoresis (CE) using alkaline buffers with or without dodecyl sulfate micelles. This paper reports the CE analysis of FITC derivatized amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine and beta-phenylethylamine in human urine using chip-based and fused-silica capillary instrumentation with laser-induced fluorescence detection. Data obtained via direct labeling of fortified urine are compared to those generated after FITC labeling of urinary extracts that were prepared by solid-phase extraction using a copolymer phase. For a urine volume of 5 mL with a "spiked amine": FITC ratio of 1:250, the latter approach was found to provide a sensitivity that is relevant for toxicological drug screening and confirmation (about 200 ng/mL urine). With direct labeling of 10 microL urine that was alkalinized and diluted for derivatization, the limit of identification was determined to be about 10 microg/mL, a value that is too high for practical purposes. Compared to fused silica capillaries, electrophoresis in microstructures is shown to provide faster separations and higher efficiencies without loss of accuracy and precision. PMID- 9870398 TI - In vivo monitoring of gabapentin in rats: a microdialysis study coupled to capillary electrophoresis and laser-induced fluorescence detection. AB - Gabapentin (GP) is a new anticonvulsant used in refractory epilepsy. Few studies have monitored the drug in vivo. We report the combination of capillary electrophoresis and laser-induced fluorescence detection (CZE-LIFD) with brain microdialysis and plasma ultrafiltration in an attempt to measure GP and offer an alternative technique for pharmacokinetic studies. We found that CZE-LIFD is capable of linearly measuring 10(-7)-10(-9) M GP in a 1 nL volume. It was also demonstrated that it is possible to monitor GP in prefrontal cortex dialysates and plasma in rats. It is concluded that the method permits in vivo monitoring of the drug in pharmacological as well as in clinical studies. PMID- 9870399 TI - Analysis of microdialysates from cancer patients by capillary electrophoresis. AB - Microdialysis (MD) is an innovative clinical technique for measuring interstitial tissue pharmacokinetics and plasma-to-tissue transfer rates of drugs in humans. However, microdialysis requires the availability of specialized analytical techniques. Capillary electrophoresis (CE), which enables concentration measurements of small volume samples, theoretically constitutes an ideal analytical technique for measuring drug concentrations in microdialysates. In the present experiments, we aimed at assessing the potential utility and limitations of CE for analysis of microdialysates in a clinical situation. Microdialysates were obtained from primary breast cancer patients who received chemotherapy including 5-fluorouracil (5-FU; 600 mg/m2). Subsequently, 5-FU concentrations were measured in tumor - and subcutaneous adipose tissue - microdialysates by CE. By combining MD and CE, complete time versus concentrations profiles could be obtained for 5-FU in the interstitial tumor space and important clinical questions could be addressed. We conclude that the combination of MD and CE leads to important and previously inaccessible information about the drug distribution process in a clinical setting. PMID- 9870400 TI - Development of a simple high-performance capillary electrophoretic method with on line mode in capillary derivatization for the determination of spermidine. AB - A new high-performance capillary electrophoretic (HPCE) method with an on-line mode in-capillary derivatization (ICD) procedure for determinations of some amines using 20 mmol/L sodium dodecyl sulfate (SDS) - 2 mmol/L o-phthalaldehyde (OPA) - 2 mmol/L N-acetylcysteine (NAC) - 20 mmol/L phosphate-borate buffer [9] has previously been shown. Although this technique offers direct fluorescence detection of free amines without any derivatization procedures before or after HPCE separation, the presence of spermidine (Spd) is difficult to detect due to low fluorescence intensity. The purpose of this study is to improve the detection sensitivity of Spd by reoptimizing this method with regard to the run buffer; the reoptimized method was applied to the determination of Spd in human plasma. To enhance the fluorescence intensity of the Spd signal, it is effective to use the run buffer in the presence of both beta-cyclodextrin (beta-CD: 8.8 mmol/L) and NAC at high concentration (16 mmol/L). By contrast, the intensity was remarkably decreased when SDS was used in the presence of beta-CD. After ultrafiltrating (UF) spiked human plasma with Spd, UF plasma was directly analyzed using the reoptimized method. Spd peak was detected and separated from the other peaks of blank plasma. The present method gave good linearity (r = 0.999), reproducibility (3.85% coefficient of variation at 5 micromol/L level; n = 10) and specificity. The detection limit and lower limit of quantitation is for 0.2 micromol/L and 1 micromol/L, respectively. PMID- 9870401 TI - Analysis of recombinant cytokines in human body fluids by immunoaffinity capillary electrophoresis. AB - An immunoaffinity capillary electrophoresis (ICE) system for rapidly quantifying recombinant cytokines in human body fluids has been developed. Cytokines within biological fluids were labeled with a red light emitting fluorochrome and injected into the capillary. Selected cytokines were captured by immobilized antibodies on the internal surface of the capillary, and held while unbound materials were purged. The cytokines were then eluted electrophoretically in acidic buffer. Individual cytokine peaks were detected by on-line laser-induced fluorescence detection coupled to a computerized fiber-optic spectrometer, and analyzed by integration of the eluted peaks. The comparison of the results of ICE to routine assays used for these cytokines demonstrates that ICE provides a fast and accurate procedure for defining these cytokines in complex biological samples. Immunoaffinity separations can be used for any material to which a specific antibody can be raised, making this procedure applicable to a wide range of molecules of biomedical interest. PMID- 9870402 TI - Determination of bupivacaine and three of its metabolites in rat urine by capillary electrophoresis. AB - A capillary electrophoretic (CE) method for the analysis of urinary extracts of the local anesthetic, bupivacaine, and its three main metabolites, desbutylbupivacaine, 3'-hydroxybupivacaine, and 4'-hydroxybupivacaine, in rat urine has been developed. The limits of detection were 0.22 microM for desbutylbupivacaine and bupivacaine, 0.15 microM for 3'-hydroxybupivacaine, and 0.16 microM for 4'-hydroxybupivacaine. The linear range was from 0.7 microM to 16.8 microM for all four compounds. Migration time and peak height reproducibilities, and extraction efficiencies were determined for all four compounds. Peak height reproducibilities (n = 5) for the overall method were improved through the use of prilocaine as an internal standard. Peak height reproducibilities were 5.6% RSD for desbutylbupivacaine and bupivacaine, and 9.9% RSD for 3'-hydroxybupivacaine and 4'-hydroxybupivacaine. Migration time reproducibilities (n = 5) were 2.4% for all compounds. Urine samples were collected from rats administered therapeutic doses of bupivacaine and extracted using a solid-phase extraction method (SPE). Separation of bupivacaine and its metabolites was achieved in 15 min. PMID- 9870403 TI - Determination of atropine in biological fluids by micellar electrokinetic capillary chromatography in the presence of strychnine and tetracaine. AB - The identification and quantitation of atropine, in whole blood and gastric contents in the presence of strychnine and tetracaine is described. This method uses liquid-liquid extraction and micellar electrokinetic chromatography (MECC). Separations are made using a 50 cm long capillary and a borate/phosphate buffer at pH 9.2 with 50 mM sodium dodecyl sulfate (SDS). Linearity was established for the three compounds between 1.0 and 100 microg/mL, using scopolamine as internal standard. The limit of detection for atropine was estimated at 0.06 microg/mL and the limit of quantitation at 0.2 microg/mL. The run time is less than 30 min. Alternate parameters are proposed to reduce the run time to under 10 min. The method was applied to a forensic post-mortem case. PMID- 9870404 TI - Serum procainamide analysis based on acetonitrile stacking by capillary electrophoresis. AB - Stacking methods are important in capillary electrophoresis (CE) to overcome the poor detection limits. Cationic drugs are difficult to stack because they tend to interact with the capillary wall. As an example of the stacking of the cationic compounds, procainamide, an anti-arrhythmic drug, is analyzed in serum by CE using an acetonitrile treatment. Serum was deproteinized with acetonitrile containing quinine as an internal standard. About 12% of the capillary volume was filled with sample and separated using an electrophoresis buffer composed of triethanolamine, 2-(N-cyclohexylamino)ethanesulfonic acid (CHES) and 20% isopropanol, pH 8.2. Both the triethanolamine and the CHES were critical for the stacking. The addition of isopropanol improved the plate number for the procainamide and decreased the interfering compounds. Procainamide, its metabolite N-acetyl procainamide, and quinine were separated in about 7 min. The CE compared well with an immunoassay method. PMID- 9870405 TI - Determination of titanocene, a new drug with anticancer potential, and its metabolism in solution by capillary electrophoresis. AB - Titanocene dichloride is one of the most promising cancerostatica of the future: nevertheless, its high activity against several tumor cells was discovered 20 years ago. Detailed knowledge of the mechanism of hydrolysis of titanocene dichloride and its stability in the infusion liquid is a prerequisite for clinical tests and for a successful application for permission as medication. Capillary electrophoresis (CE) was used to observe the hydrolysis behavior of titanocene dichloride in aqueous solutions. The hydrolysis products were separated in a 20 mM phosphate buffer, pH 6, and in a 20 mM malic acid buffer, pH 3. Up to five hydrolysis products were obtained. A significant influence of the sample preparation (pH, isoionic additives) on the hydrolysis rate was observed. The hydrolysis products were characterized by the UV scan and the element selective particle-induced X-ray emission (PIXE) detection technique. The results obtained correspond with the hydrolysis mechanism described in the literature. The determination of free titanocene dichloride in human plasma failed due to the high affinity of the plasma proteins for this compound. PMID- 9870406 TI - Capillary ion analysis of lithium concentrations in biological fluids and tissues of Poecilia (teleost). AB - Capillary ion analysis (CIA) is a form of capillary electrophoresis that uses the differential electrophoretic mobility of ions to perform a separation of an ionic mixture. Application of this technique for detection of lithium concentrations in plasma and tissues of Poecilia was the purpose of this investigation. CIA was performed using a 75 microm ID x 60 cm length fused-silica capillary and a run electrolyte of 67.7 mg hydroxyisobutyric acid (HIBA), 52.8 mg 18-crown-6-ether and 64 microL UV-CAT-1 reagent (4-methylbenzylamine) in a volume of 100 mL water (18 (M)omega) with a voltage of 20 kV using ultraviolet absorption detection at 214 nm. Migration times were: potassium, 2.98 min; calcium, 3.48 min; sodium, 3.60 min; barium (internal standard), 4.15 min and lithium, 4.26 min. Lithium and barium migration times were stable and reproducible. Correlation coefficients (r) between peak area ratios of lithium/barium for concentrations ranging from 0.1 to 2.0 mM were from 0.976 to 0.996. Coefficients of variation (CV) for lithium concentrations ranged from 4.07 to 15.71% between days and 4.38 to 7.76% within day. Application of this methodology for determination of lithium concentrations in the plasma, brains and livers of fish dosed with lithium for 23 days are presented. CIA is applicable to analysis of lithium concentrations in biological fluids and tissues of fish. PMID- 9870407 TI - Separation of etoposide phosphate and methotrexate by capillary zone electrophoresis using UV detection with a high sensitivity cell. AB - Etoposide phosphate and methotrexate are important anti-tumor chemotherapeutic agents. Our previously presented capillary zone electrophoresis (CZE) method using a high sensitivity cell (Z-cell) for quantitative analysis in biological media (urine, plasma) showed good precision and accuracy. The present results show that the investigation using a capillary with high sensitivity cell led to an approximately 10-fold improvement of the detection limit compared to standard capillaries. Plasma and urine samples were analyzed by using a calibration curve for drug concentrations between 0.1 and 100 microg/mL. Good detection limits and good relative standard deviations of the migration times and of the peak areas were observed in these experiments. PMID- 9870408 TI - Analysis of isoquinoline alkaloids in medicinal plants by capillary electrophoresis-mass spectrometry. AB - The technique of capillary electrophoresis - mass spectrometry (CE-MS) was applied for determination of isoquinoline alkaloids in crude methanolic extracts of medicinal plants. For the CE separations ammonium formate buffer solutions (70 or 100 mM, pH 3.0 or 4.0) containing 10% methanol or 20-60% acetonitrile as additives were used. The applied voltage was 25 kV, the thermostating temperature was kept constant at 25 degrees C. Coupling with the mass spectrometer was performed via an atmospherical pressure ionization (API) interface and the electrospray ionization technique (ESI). As sheath liquid 5 mM formic acid in acetonitrile at a flow rate of 3 microL/min was used. The spray voltage was 4.5 kV and the temperature of the heated capillary was chosen to be 200 degrees C. Detection in the positive ionization mode resulted in mass spectra showing either the molecular ions [M]+ or the protonated molecular ions [M+H]+. The presented method allows detection and identification of isoquinoline alkaloids in crude methanolic extracts of medicinal plants as Eschscholzia californica CHAM. (Papaveraceae), Hydrastis canadensis L. (Ranunculaceae), Berberis vulgaris L. (Berberidaceae), Jateorhiza palmata (LAM.) MIERS (Menispermaceae) and Chelidonium majus L. (Papaveraceae). PMID- 9870409 TI - Optimized determination of carbohydrate-deficient transferrin isoforms in serum by capillary zone electrophoresis. AB - Carbohydrate deficient transferrin (CDT) is one of the most reliable markers of chronic alcohol abuse. It consists of a group of minor isoforms of human transferrin (the main iron transport serum protein) deficient in sialic acid groups (asialo, monosialo and disialo) with a pI > 5.7, while the main isotransferrin (tetrasialo) has a pI of 5.4. The aim of the present work was to develop a capillary electrophoretic method to determine CDT in serum, suitable for routine use as a confirmatory technique of the current screening methods based on immunoassays. Serum samples (0.5 mL) were saturated with iron by incubation with 10 mM FeCl3 (9 microL) and 500 mM NaHCO3 (12 microL) for 30 min, then diluted 1/10 in water and injected by positive pressure (0.5 psi for 10 s). Separation was performed with a capillary zone electrophoretic method using bare fused-silica capillaries (20 microm ID, 37 cm in length) and a buffer composed of 100 mM sodium tetraborate adjusted with boric acid to pH 8.3. Applied voltage was 10 kV and temperature 25 degrees C. Detection was by UV absorption at 200 nm wavelength. Under the described conditions, asialo-, monosialo-, disialo-, trisialo- and tetrasialo-transferrin were separated in human serum. The limit of detection (signal-to-noise ratio of 2) was about 0.3% for disialo-transferrin, and 0.4% of trisialo-transferrin, expressed as percentages of the terasialo transferrin peak area. Relative standard deviations (RSD) of absolute migration times were < 1%, while RSD of relative migration times (on the basis of tetrasialo-transferrin) were < 0.1%. Intra-day and day-to-day peak quantitation precision studies showed RDS ranging from 4 to 9% and from 13 to 24% for disialo- and trisialo-transferrin, respectively. The results from 30 control subjects, including social drinkers, and 13 alcoholics showed disialo- and trisialo transferrin significantly increased in patients by a factor of about 4.5 (P < 0.0001). PMID- 9870410 TI - Monoclonal antibody binding affinity determined by microchip-based capillary electrophoresis. AB - The affinity constant of a monoclonal antibody to fluorescently labeled bovine serum albumin (BSA) was measured in diluted mouse ascites fluid using a microfluidic chip to perform affinity capillary electrophoresis. Borofloat glass based devices could be used repeatedly with samples for many months. On-chip separations were performed in less than 60 s, and 30-60 s was required for manual sample exchange. The change in peak height for BSA with increasing BSA/anti-BSA concentration ratio was used to determine concentration changes in bound and free BSA. A Scatchard plot analysis gave an affinity constant (more exactly the intrinsic association constant) of 3.5+/-0.6 x 10(7) M(-1) for a 1:1 stoichiometric ratio. Two affinity complexes were separated. One complex was identified by the Scatchard method as having a 1:1 stoichiometric ratio. The other complex is proposed to have a stoichiometry with an excess of anti-BSA to BSA, most likely (anti-BSA)2-BSA, on the basis of a faster migration time than the 1:1 complex, a decrease in the amount of this complex with increasing [BSA], and predictions of theoretical models for multi-valent antigens. Potential applications of microchip-based devices in affinity measurements are discussed. PMID- 9870411 TI - Charged chelate-capillary electrophoresis of endogenous corticosteroids. AB - Separation of endogenous 17- or 18-hydroxylated corticosteroids (of the 21 hydroxylated 4-pregnen series) as charged chelates in capillary electrophoresis with borate as the ligand is demonstrated. Aldosterone, 18-hydroxycorticosterone, 18-hydroxy-11-deoxycorticosterone, cortisone, cortisol, and 11-deoxycortisol are separated and resolved by 400 mM borate buffer at pH 9.0. Separation characteristics of the corticosteroid charged chelates were examined by varying the separation buffer borate concentration, pH, ionic strength, and addition of organic modifiers. The borate ion [B(OH)4]- is identified as the critical buffer component. Corticosteroids chelate borate with proximal hydroxyls composed of either the 17- or 18-hydroxyl in combination with the 21-position hydroxyl. Corticosteroid/borate chelation as indicated by CE results is corroborated with 11B-nuclear magnetic resonance (11B-NMR) spectra. Chelation is a readily reversible process, with the strength of the resultant chelate, as opposed to the charge-to-mass ratio, predominantly determining analyte mobility in charged chelate - capillary electrophoresis (CC-CE). PMID- 9870412 TI - FDG-PET predictors of response to behavioral therapy and pharmacotherapy in obsessive compulsive disorder. AB - In subjects with obsessive-compulsive disorder (OCD), lower pre-treatment metabolism in the right orbitofrontal cortex (OFC) and anterior cingulate gyrus (AC) has been associated with a better response to clomipramine. We sought to determine pre-treatment metabolic predictors of response to behavioral therapy (BT) vs. pharmacotherapy in subjects with OCD. To do this, [18F]fluorodeoxyglucose positron emission tomography scans of the brain were obtained in subjects with OCD before treatment with either BT or fluoxetine. A Step-Wise Variable Selection was applied to normalized pre-treatment glucose metabolic rates in the OFC, AC, and caudate by treatment response (change in Yale Brown Obsessive-Compulsive Scale) in the larger BT group. Left OFC metabolism (normalized to the ipsilateral hemisphere) alone was selected as predicting treatment response in the BT-treated group (F = 6.07, d.f. = 1,17, P = 0.025). Correlations between normalized left OFC metabolism and treatment response revealed that higher normalized metabolism in this region was associated with greater improvement in the BT-treated group (tau = 0.35, P = 0.04), but worse outcome (tau = -0.57, P = 0.03) in the fluoxetine-treated group. These results suggest that subjects with differing patterns of metabolism preferentially respond to BT vs. medication. PMID- 9870413 TI - Magnetic-resonance morphometry in patients with major depression. AB - Magnetic-resonance morphometry performed on 72 patients with major depression compared with 38 control subjects replicated previously reported, statistically significant reductions in the volumes of the caudate (P < 0.03) and putamen (P < 0.05) in depressed patients. Borderline statistical significance was observed for whole-brain (P < 0.07) and frontal volume (P < 0.10) in a subsample of 32 patients matched on age and sex with 32 control subjects, whereas statistical significance was observed for the full sample (P < 0.007 and P < 0.03, respectively). Chronological age was related to volume of the frontal lobes (P < 0.0002), caudate (P < 0.0001), putamen (P < 0.008), thalamus (P < 0.002), cerebellum (P < 0.007), lateral ventricles (P < 0.0001), and ratios of [whole brain]/[whole brain + cerebrospinal fluid (CSF)] (P < 0.0001) and [frontal]/[frontal + CSF] (P < 0.0001). Age of first depressive episode was related to putamen volume after accounting for chronological age (R2= 0.16, P < 0.005), and a correlation of 0.26 (P < 0.04) was observed between caudate volume and global mental status. Results are in accord with previous reports of basal ganglia abnormalities in depressed patients and support the role of subcortical structures in mediating affective disorder. PMID- 9870414 TI - Single-voxel proton magnetic resonance spectroscopy of the pons and cerebellum in patients with schizophrenia: a preliminary study. AB - Magnetic resonance spectroscopy (MRS) is a non-invasive functional imaging technique that can measure various brain tissue metabolites such as N acetylaspartate (NAA), choline (Cho), creatine-phosphocreatine (Cr), myo-inositol (mI) and other metabolites. Morphological studies have indicated the pons and cerebellum as possible sites of abnormal functioning in schizophrenic patients. This study examines schizophrenic patients for the presence of abnormalities in proton MRS (1H-MRS) measured metabolites in two regions of the posterior fossa. Twelve schizophrenic patients and eight non-schizophrenic control subjects were studied by measuring the ratios of NAA/Cr, Cho/Cr and mI/Cr from 1H-spectra obtained from the pons and right or left cerebellum using an integrated MRI/MRS protocol. Spectra were obtained from a voxel in the pons and voxels from the left and/or right lateral cerebellum. Data were analyzed in the absorption mode and fitted to Lorentzian lineshapes using a Marquart algorithm. Significantly lower NAA/Cr ratios were found in the pons of schizophrenic patients than in the control subjects, but not in the cerebellum. This study is the first to measure brain tissue metabolites using 1H-MRS in the pons and cerebellum of schizophrenic patients. Significant alterations of 1H-MRS metabolites may suggest the involvement of the posterior fossa as a part of the pathological substrate underlying schizophrenia. PMID- 9870415 TI - Multimodal imaging of residual function and compensatory resource allocation in cortical atrophy: a case study of parietal lobe function in a patient with Huntington's disease. AB - In a case of Huntington's disease (HD) with dementia and pronounced parieto frontal atrophy, the functional state of the affected regions was investigated using functional magnetic resonance imaging (fMRI) and fluorodeoxyglucose positron emission tomography (FDG-PET). It was observed that although parietal areas showed extensive atrophy and reduced resting glucose metabolism, the patient performed with similar accuracy but with longer response time in a visuospatial task compared with healthy control subjects. At the same time, the blood oxygen level-dependent (BOLD) fMRI signal in these areas, which are involved in visuospatial processing, showed a similar task-dependent modulation as in control subjects. The signal amplitude (signal percent change) of the task dependent activation was even higher for the HD patient than in the control group. This residual functionality of parietal areas involved in visuospatial processing could account for the patient's performance in the task concerned, which contrasted with his poor performance in other cognitive tasks. The increased percent-signal change suggests that a higher neuronal effort was necessary to reach a similar degree of accuracy as in control subjects, fitting well with the longer reaction time. We propose that fMRI should be considered as a tool for the assessment of functionality of morphologically abnormal cortex and for the investigation of compensatory resource allocation in neurodegenerative disorders. PMID- 9870416 TI - Decreased duration and altered topography of electroencephalographic microstates in patients with panic disorder. AB - The topography and temporal sequence of scalp electrical fields were analyzed by adaptive segmentation of the continuous electroencephalogram (EEG) in 27 patients with panic disorder and 28 control subjects during rest phases and during the viewing of a neutral (mushroom) or an emotionally relevant (casualty) picture. The results indicate decreased duration of brain microstates in panic patients during all conditions. Comparison of the resting phases with the viewing conditions revealed a significant acceleration of EEG microstates in both the patients and the control subjects. Patients and control subjects differed in the topography of the fields during rest: control subjects showed a left anterior/right-posterior orientation, while panic patients showed a predominantly right-anterior/left-posterior orientation. Neither group displayed any topographic changes when viewing the mushroom picture. However, when viewing the anxiety-specific casualty picture, panic patients shifted fields in a different way than did control subjects. Centroid topography does not permit clear localization of the cortical generators. It is concluded that panic patients show a generally increased cortical activation compared with healthy control subjects, and activate different neuronal arrays when viewing an anxiety-specific stimulus. PMID- 9870417 TI - The epidemiology of diabetes mellitus. AB - Insulin-dependent diabetes mellitus (IDDM) develops predominantly in children and young adults, but may appear in all age groups. The incidence of IDDM differs greatly among populations, with Finland and Sardinia showing the greatest incidence rates (approximately 30-35% of cases annually per 100000 children up to age 14 years) and oriental populations showing the lowest rates. IDDM is diagnosed more frequently in the winter months. The major genetic susceptibility to IDDM is linked to the HLA complex on chromosome 6. These genetic backgrounds interact with environmental factors (possibly certain viruses, foods and climate) to initiate the immune-mediated process that leads to beta-cell destruction. Non insulin dependent diabetes (NIDDM) is the most common form of diabetes. The prevalence of NIDDM varies enormously from population to population. The greatest rates have been found in Pima Indians. The major environmental factors identified as contributing to this form of diabetes are obesity and reduced physical activity. NIDDM shows strong familial aggregation in all populations and is clearly the result of an interaction between genetic susceptibility and environmental factors. Before NIDDM develops, insulin concentrations are high for the degree of glycaemia and of obesity, reflecting the presence of insulin resistance. As insulin resistance worsens, glucose levels increase, with the appearance of glucose intolerance and, finally, of NIDDM, when insulin response cannot compensate for insulin resistance. PMID- 9870418 TI - The drama of the continuous increase in end-stage renal failure in patients with type II diabetes mellitus. AB - Type II diabetes mellitus has become the leading cause of end-stage renal failure in many countries of Western Europe. In all European countries, even in those with a relatively low prevalence of diabetic nephropathy, the number of patients with type II diabetes mellitus admitted for renal replacement therapy has recently increased continuously. Survival and medical rehabilitation of patients with type II diabetes on renal replacement therapy is significantly worse than in non-diabetic patients. It is obvious that in order to stem the tide, intense efforts are necessary (i) to inform the medical community about the renal risk of type II diabetes and the striking effectiveness of preventive measures, (ii) to provide better care for diabetic patients, and (iii) to reduce the high prevalence of diabetes in the population by modification of the Western life style. PMID- 9870419 TI - Diabetic nephropathy: from micro- to macroalbuminuria. AB - This brief review will focus on the major factors leading to incipient diabetic nephropathy (i.e. microalbuminuria) progressing to overt nephropathy (i.e. macroalbuminuria) and particularly on the role of glycaemic control and hypertension. Both experimental and cohort studies support the role of hyperglycaemia in the development of diabetic nephropathy. Some recent long-term interventional studies in microalbuminuric patients show conflicting results regarding the role played by good metabolic control in reducing the incidence of overt nephropathy. However, strict metabolic control, which is fundamental in normoalbuminuric patients, is of little use even in microalbuminuric patients. In general, levels of glycosylated haemoglobin less than two standard deviations above the upper normal range, commonly <7.5-8%, seem to protect patients from developing nephropathy. The results of many cross-sectional studies have shown that the progression of renal damage regularly is accompanied by arterial hypertension both in insulin-dependent diabetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM). Many long-term interventional studies have been performed in order to understand the effect of antihypertensive treatment on the incidence of proteinuria in both normotensive and hypertensive patients with IDDM or NIDDM. These data show a marked effect of antihypertensive therapy in preventing the onset of overt nephropathy, and suggest the superiority of angiotensin-converting enzyme (ACE) inhibitors. We believe that optimal blood pressure values are approximately 120/70-75 mmHg in younger patients and 125 130/80-85 mmHg in older patients. In conclusion, antihypertensive treatment, ACE inhibitors per se and possibly strict metabolic control reduce the development of nephropathy, thus playing a striking role in the secondary prevention of renal failure. PMID- 9870420 TI - Progression of renal failure in diabetic nephropathy. AB - The onset of renal damage in diabetes mellitus may be influenced by several factors which largely result from genetic predisposition, hereditary factors and the early appearance of microalbuminuria and/or systemic hypertension. Most of these factors are also implicated in the progression of nephropathy from microalbuminuria to overt proteinuria and to end-stage renal failure (ESRF). Over the last few years, the role of hyperglycaemia has emerged as critical in mediating the progressive renal damage in diabetes. However, hyperglycaemia leads to increased formation of glycated proteins which may act as promoters of progression by localizing in renal tissue. In addition, hyperglycaemia may have a synergistic effect with some other risk factors, such as growth factors and the renin angiotensin system, in accelerating renal deterioration. PMID- 9870421 TI - Molecular biology of diabetic glomerulosclerosis. AB - Diabetic nephropathy is one of the leading causes of renal failure in Western countries, where diabetic patients account for nearly half of all patients on haemodialysis. Progressive expansion of the mesangial matrix, and thickening of the glomerular and tubular basement membranes without signs of major cell proliferation are hallmarks of human and experimental diabetic nephropathy. These lesions eventually lead to glomerular fibrosis, a central pathological feature in many human acute and chronic kidney diseases, which progressively destroys the renal filtration unit, and may finally cause renal failure. Indeed, structure function relationship studies have shown that mesangial matrix expansion is strongly related to the clinical manifestation of diabetic nephropathy. PMID- 9870422 TI - Renovascular disease in diabetes mellitus: treatment by percutaneous transluminal renal angioplasty. AB - BACKGROUND: Diabetes mellitus is an important cause of end-stage renal failure (ESRF). Although classic diabetic nephropathy accounts for the majority of patients reaching ESRF, renovascular disease, which is frequent in such patients, plays an increasingly important role. Percutaneous transluminal renal angioplasty (PTRA) has been proven to be an efficacious measure for renal revascularization. METHODS: Ninety-nine patients with diabetes mellitus and renal artery stenosis, corresponding to 16.6% of the entire population of diabetic patients, were treated by PTRA or with the Palmaz-Schatz stent in our clinic. Technical success was achieved by PTRA in 92/99 patients, in 10 patients a Palmaz-Schatz stent was implanted. RESULTS: Hypertension was cured in eight and improved in 44 patients. In 47 patients, there was no impact on blood pressure. An improvement in renal function was evident 1 month after PTRA in 8/27 patients. A further improvement occurred in another four patients after 6 months. The re-stenosis rate was 22% after 5 years. Serious complications occurred in seven patients (one patient required surgery and two patients had regular dialysis treatment). CONCLUSIONS: Renovascular disease is an important cause of ESRF in diabetic patients. PTRA is a valid tool to revascularize renal artery stenosis and improve blood pressure control and renal function both in diabetic and non-diabetic patients. PMID- 9870423 TI - Short-term outcome of diabetic patients in renal replacement therapy. AB - BACKGROUND: Diabetic nephropathy or diabetes-related nephropathies represents one of the most relevant causes of renal failure in recent years. This complex pathological picture becomes particularly severe as time elapses and after starting renal replacement therapy (RRT). METHODS: In an attempt to investigate the evolution of the major clinical features, a retrospective study was carried out on a cohort of 76 diabetic patients on RRT. Sixty-five have been treated by haemodialysis (HD) and 11 by peritoneal dialysis (CAPD), for at least 1 year. In these patients change in modality of treatment, metabolic control, cardiovascular, and ophthalmological complications, peripheral neuropathy, state of vascular access, and intradialytic complications were surveyed at initiation and after 1 year of treatment. A modified Karnofski's score was utilized, to evaluate the degree of rehabilitation. The comparison of prevalence was evaluated, using Student's t-test for paired samples. RESULTS: After 1 year, 11 patients on CAPD remained on the same type of treatment. Out of 65 patients on standard bicarbonate HD, 11 were moved to acetate free biofiltration, two to paired filtration dialysis and one to haemofiltration. A worsening in arrhythmias was recorded with an increased prevalence from 25.0 to 35.0% (n.s.), and one more patient (15 vs 16 and 19 vs 20 respectively) experienced ischaemic cardiomyopathy and cerebrovascular insufficiency. Hypertension showed a significant improvement (72 vs 42, P<0.01). Nausea and vomiting, hypotensive episodes, and muscular cramps were more frequently observed. A worsening in patient's welfare was also recorded but without statistical significance. CONCLUSIONS: This clinical evaluation even if retrospective and lasting 1 year, may suggest that RRT does not per se represent a cause of the development and progression of the major complications related to diabetic disease. PMID- 9870424 TI - The effect of metabolic control on development and progression of diabetic nephropathy. AB - The progressively growing number of patients with end-stage renal failure (ESRF) associated with diabetes mellitus and requiring renal replacement therapy (RRT) stimulated both nephrologists and diabetologists to investigate the mechanisms linking hyperglycaemia to diabetic renal failure and to set up measures to prevent the onset and slow the progression of diabetic nephropathy. Over the last few decades, a large number of studies have investigated both the incidence of diabetic nephropathy and the relationship between metabolic control and the development of diabetic nephropathy. Chronologically, the first type of diabetes and diabetic nephropathy to be studied was type I, and it is only in recent years that metabolic control has been proven to be a contributor to the development of nephropathy in such patients. Recently, the DCCT demonstrated that metabolic control in the prealbuminuric phase was effective in reducing the incidence of microalbuminuria, even if it was unable to reduce the incidence of overt proteinuria in patients with type I diabetes and established proteinuria. On the other hand, in type II diabetes, the number of studies demonstrating a favourable effect of metabolic control on onset and progression of diabetic nephropathy is only slightly greater than those that failed to show a favourable effect. This feature may suggest that in type II patients, genetic and ethnic differences, nutritional aspects, lifestyle and other confounding factors may play a relevant role in the course of the disease. However, the trials performed and the retrospective analyses generally agree that glycated haemoglobin two standard deviations greater than the mean is related to a worsening in progression of diabetic nephropathy and to an enhanced risk of other complications. In general, a glycated haemoglobin < or =8% seems advisable. Moreover, in both type I and type II, greater emphasis should be placed on the major risk factors such as hypertension, smoking habits and hyperlipidaemia. PMID- 9870425 TI - Efficacy of antihypertensive therapy in decreasing renal and cardiovascular complications in diabetes mellitus. AB - The mechanism underlying the pathogenesis of microangiopathy and macroangiopathy in diabetes mellitus is hypothesized to be chronic hyperglycaemia. However, the values of blood glucose at which chronic diabetic complications develop at the renal and cardiac level are quite different. It is not clear whether this is due to different responses of kidney and heart tissues to the metabolic challenge of diabetes, or to the simultaneous role of other mechanisms contributing differently to the pathogenesis of chronic diabetic complications in renal and cardiac tissues. One of these mechanisms could be the simultaneous occurrence of arterial hypertension along with hyperglycaemia in diabetic patients. We reviewed the available evidence in the recent medical literature and provide information on the relationships between hyperglycaemia and cardiovascular and renal complications in insulin-dependent diabetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM). The majority of reports indicate that the values of blood glucose appearing to be at threshold level for the development of cardiovascular complications are significantly lower than those determining renal complications (5.4 vs 10.0 mmol/l blood glucose concentrations 2 h after an oral glucose tolerance test). This was the case both in cross-sectional and prospective studies and also in intervention studies aimed at decreasing blood glucose concentrations by using strict metabolic control methods (The Diabetes Control and Complications Trial Research Group), which succeeded in delaying the rate of occurrence of microangiopathic complications at the kidney and retinal level but not so effectively at the cardiac level. Therefore, alternative therapeutic or supplementary strategies to blood glucose control should be adopted in diabetes to prevent diabetic complications. To date, the most effective approach, from our point of view, is antihypertensive therapy in order to prevent end-stage renal disease. We extensively reviewed the available literature which reported comparisons between angiotensin-converting enzyme inhibitors (ACE inhibitors) and calcium channel blockers (CCBs) in the treatment of arterial hypertension in diabetes. Intensified antihypertensive therapy achieving a blood pressure level below 130/85 mmHg has been shown to be useful in decreasing the rate of occurrence of chronic diabetic complications in diabetes mellitus. Both ACE inhibitors and CCBs have been shown to significantly improve the course of renal function in diabetic patients with incipient and overt nephropathy. PMID- 9870426 TI - Dietary treatment of diabetic nephropathy with chronic renal failure. AB - Thirty-two patients with diabetes mellitus (22 IDDM and 10 NIDDM, 21 males and 11 females, age 44+/-11.8 years) were followed for 5.2+/-3.8 years after the onset of chronic renal failure, with the aim of evaluating the effect of low protein diets on the rate of decline of the residual renal function. During the 1.8+/-1.6 year follow-up period on free or uncontrolled low protein diet the mean rate of decline of creatinine clearance was 0.9+/-0.6 ml/min/month, significantly greater than that observed during 3.7+/-3.1 years on low or very low protein diets. The reduction of protein intake was followed by a significant decrease in daily urinary protein loss. A better glycaemic control was obtained on the low protein diet, and the daily insulin requirement decreased. The anthropometry, as well as the serum concentrations of rapid turnover proteins, did not change, in spite of the low or very low protein dietary supply for a long duration. The values of mean arterial pressure were quite similar during the follow-up period on free or uncontrolled low protein diet and during the study period on the low protein diet. A good compliance with reduced dietary intake (as demonstrated by the measurement of the daily urea excretion) was obtained in a large number of patients. In conclusion, our study confirms the protective effect on the residual renal function of low protein diets in IDDM and NIDDM patients with chronic renal failure due to diabetic nephropathy, in the absence of any sign of protein malnutrition. PMID- 9870427 TI - The treatment of diabetic end-stage renal disease with peritoneal dialysis. PMID- 9870428 TI - Complications of pancreas transplantation in an initial experience of a transplant programme. PMID- 9870429 TI - Cardiovascular risk factors, smoking and kidney function. PMID- 9870430 TI - Cardiovascular abnormalities in ageing and in uraemia--only analogy or shared pathomechanisms? AB - The analogies between the effects of ageing and of uraemia on the function and the structure of central elastic arteries and of the heart are striking. Qualitatively similar changes are seen in pulse contour, pulse wave velocity, and impedance and also similar structural abnormalities with wall thickening, diminished elastin, and increased collagen content. The altered 'Windkessel' function of central arteries in age and uraemia is one factor contributing to left ventricular hypertrophy. PMID- 9870431 TI - Acute interstitial nephritis in the elderly: a report from the UK MRC Glomerulonephritis Register and a review of the literature. PMID- 9870432 TI - Vasomotor nephropathy in the elderly. PMID- 9870433 TI - Acute renal failure of medical type in an elderly population. AB - One hundred and nine unselected patients with Acute Renal Failure (ARF) of medical aetiology were hospitalized at the Nephrological Unit of Policlinico University Hospital (Modena) during a 30-month period. ARF was considered as a rapid increase of serum creatinine > 2mg/dl over the baseline level or the doubling of pre-existing value in chronic renal failure. Mean age of patients was 67+/-17 years and median age was 72; 64.2% needing dialytic treatment. Four main causes of ARF were identified: 33 patients had reduced renal perfusion by dehydration, hypotension etc.; 20 multifactorial aetiology; 14 biopsy investigated renal parenchymal diseases and 39 had drug-related acute renal failure (D-ARF). The clinical outcome was significantly worse in elderly patients as regard mortality (P < 0.02), chronic dialytic treatment (P < 0.04) and complete recovery (P < 0.004). The mean age of D-ARF patients was significantly greater than remaining ARF patients (72.6+/-12.8 vs 63.2+/-18.5: P < 0.004. Nonsteroidal antiinflammatory drugs (NSAIDs) and ACE-inhibitors (Ace-i) caused ARF in 24 and 8 patients respectively. Elderly age, vascular disease and monoclonal gammopathy represented the main risk factors and were significantly more frequent in D-ARF patients (P<001, <0.01, <0.04 respectively). Our data confirm the high susceptibility of ageing kidneys to nephrotoxic damage caused by drugs affecting glomerular autoregulation by microvascular mechanisms. Greater attention to renal changes in ageing and an increased dissemination of preventative measures among nephrologists, could reduce the incidence of these serious and potentially lethal diseases. PMID- 9870434 TI - Renal impairment in myeloma: time for a reappraisal? AB - It is generally considered that a patient with myeloma who also has established renal impairment is unlikely to do well. While this is sometimes the case, analysis of recent data shows: (a) of 2768 patients in the MRC database for the fourth to the sixth trials, 10/163 with serum creatinine 300-600 microm/l and 20/89 with serum creatinine 600 micro/l at presentation had renal failure as a recorded presenting feature, whatever the renal function, the most common presenting feature was bone pain; (b) that many patients have persisting evidence of reduced renal function yet survive for more than the median time of 36 months which applies to myeloma generally. Patients with renal impairment, especially those whose myeloma is brought to plateau by chemotherapy, should be assessed thoroughly for evidence of other than myeloma induced renal damage as both renovascular disease and prostatic obstruction in males are common in the elderly population at risk. PMID- 9870435 TI - Human brain amyloidoses. PMID- 9870436 TI - Neuromuscular complications of kidney diseases. PMID- 9870437 TI - Genetic kidney diseases in the elderly. PMID- 9870438 TI - Clinical biology of artificial organ substitution. PMID- 9870439 TI - Which level of cytokine production is critical in haemodialysis? AB - Monocyte activation with cytokine production is a well known event in the course of dialysis treatment but its relation to symptoms of haemodialysis or long-term pathological changes in chronic dialysis patients is still under discussion. Cytokine production depends on the balance between inducers and inhibitors while effects rely on the peculiar uraemic environment and cell metabolism. 'Foot prints' for monocyte activation have been found, but no marker for clinical symptoms has been demonstrated clearly. In this scenario it is almost impossible to link a specific symptom to a definite stimulus such as dialysate microbial contamination or membrane complement generation. The topics discussed in this paper include cytokines synthesis modulation factors, levels in haemodialysis patients, and results of finding markers of clinical relevance. Special attention is paid to microbial contamination of dialysis fluid with analysis of cytokine inducing substances in commercial sterile solutions. Data on cytokine synthesis and activity in the aged are also discussed, with special regard to the haemodialysis setting. PMID- 9870440 TI - Dialysis-related amyloidosis: importance of biocompatibility and age. AB - The histological prevalence of dialysis-related amyloidosis (DRA) is much greater than suspected on clinical grounds: one-third of patients are affected after less than 4 years on haemodialysis (HD) and over 90% after more than 7 years HD. Risk factors include the time on dialysis, the type of HD membrane, and the age of the patient at onset of dialysis. The protective effect of high-flux membranes such as AN69 probably results mainly from the greater clearance of beta2 microglobulin. Other potential but more controversial explanations include a protective influence on residual renal function, a lower stimulation of beta2 microglobulin synthesis or release, or a beneficial influence on advanced glycosylation end (AGE) products. The higher risk of DRA in older patients has recently been suggested to result from an age-related AGE-modification of osteoarticular collagen. The best prevention and treatment of DRA is successful renal transplantation. In patients unsuitable for transplantation, high flux membranes such as AN69 should be used from the start of dialysis. Palliative treatment includes analgesics, low dose prednisone in severe cases, and surgical treatment of complications. PMID- 9870441 TI - Epidemiology and demography of treated end-stage renal failure in the elderly: from the European Renal Association (ERA-EDTA) Registry. PMID- 9870442 TI - The elderly dialysis patient: management of the hospital stay. AB - Our data show that survival times in elderly patients can be achieved, which justify the strain of the therapy on the patient, the medical effort and financial expense. It is therefore not justifiable to withhold dialysis from a person who requires it on the basis of age. It is also wrong to reduce the therapeutic endeavours to a minimum and describe these then as 'kinder' to the patient. It is to be feared that such a treatment regimen leads to a shorter survival time and simultaneously to a decreased quality of life. Particularly in view of the person's advanced age, it would seem that we are obligated to treat them with the highest quality care so that the remaining time is not only extended, but is filled with life. PMID- 9870443 TI - Lessons from 494 permanent accesses in 348 haemodialysis patients older than 65 years of age: 29 years of experience. AB - Currently, patients older than 65 years of age constitute more than 42% of all new enrolments for dialytic treatment in the USA and Italy. Most of these patients are treated by in-centre haemodialysis (HD), with problems connected to vascular access. Personal experience of 494 new vascular accesses in 348 'difficult' HD-patients older than 65 years over 29 years showed the best results from 221 elbow fistulas in comparison with 32 forearm fistulas (78% vs 57.2% at 3 years; P < 0.05). Among various vascular substitutes, the homologous saphenous vein (HSV) graft, alone or mixed (MX) gave the best secondary patency in comparison with other organic-semiorganic (OSO) or synthetic graft (SYN) angioaccesses with values of 59.4% for HSV, 66.3% for MX, 21.9% for OSO, and 38.6% for SYN grafts, respectively at 3 years. PMID- 9870444 TI - Vascular access for haemodialysis: from surgical procedure to an integrated therapeutic approach. AB - During the past 10 years the type of vascular access for haemodialysis procedures have changed markedly in our centre: more elbow AV fistulae and more central venous catheters are now used. Nevertheless, early referral to nephrologists and availability of central venous catheters and peritoneal dialysis allow elderly people to be admitted for dialysis treatment. Since vascular access for haemodialysis plays a key role in patient well-being, it is mandatory to apply quality assurance criteria to vascular access for haemodialysis surgery. Based on the results of a national survey, in Italy this policy is still in its early stages: monitoring of vascular access differs amongst centres, interventional radiology is used in a differing way, planning of vascular access for haemodialysis in pre-dialysis patients often remains an unsolved problem. According to our initial experience, we propose the use and validation of a quality-index [(minimum success rate) in elective vascular access for haemodialysis surgery], allowing accreditation of a department and a single surgeon for access management. Prevalence of central venous catheters at first dialysis of chronic renal failure patients is also proposed to evaluate the efficiency in access planning. Better knowledge of vascular access management by different teams could eventually lead to definition of guidelines for this 'Cinderella of dialysis'. PMID- 9870445 TI - Permanent twin catheter: a vascular access option of choice for haemodialysis in elderly patients. PMID- 9870446 TI - Psychological and social problems of dialysis. PMID- 9870447 TI - Angiotensin II-complexities beyond AT1 and AT2 receptors. PMID- 9870448 TI - ACE knockout mice--lessons for adult nephrology. PMID- 9870449 TI - Sodium proton exchanger (NHE) isoforms--potential relevance to hypertension and its complications. PMID- 9870450 TI - Glucose production by the human kidney--its importance has been underestimated. PMID- 9870451 TI - Methicillin-resistant Staphylococcus aureus (MRSA) infection in glomerulonephritis--a novel hazard emerging on the horizon. PMID- 9870452 TI - Management of the patient with Behcet's disease. AB - Behcet's disease is a vasculitic disorder in which the aetiopathogenetic pathway has not yet been clarified. Many organs and systems may be affected in BD. The causes of the wide-ranged clinical spectrum and the variable severity of involvement, including the kidneys remain to be defined. Behcet's disease should also be considered in the differential diagnosis of AA amyloidosis as well as vasculitic connective-tissue disorders. Specific laboratory tests for the diagnosis and follow up are needed. Only clarification of the aetiopathogenesis of BD can lead to better treatment options. PMID- 9870453 TI - Pregnancy and end-stage renal disease--past experience and new insights. PMID- 9870454 TI - Is renal failure caused by primary hypertension? Why does the controversy continue? PMID- 9870455 TI - Renaissance of the batch method? PMID- 9870456 TI - Prophylaxis of cytomegalovirus infection in renal transplantation. PMID- 9870457 TI - The global burden of disease and the challenge of invisibility. PMID- 9870458 TI - Inward rectifier renal potassium channel (ROMK), the low-conductance channels for potassium secretion. PMID- 9870459 TI - Dialysis hypotension: do we see light at the end of the tunnel? PMID- 9870460 TI - The Normal Haematocrit Trial in dialysis patients with cardiac disease: are we any the less confused about target haemoglobin? PMID- 9870461 TI - The first clinical and epidemiological programme on renal disease in Bolivia: a model for prevention and early diagnosis of renal diseases in the developing countries. AB - BACKGROUND: The prevalence and incidence of renal diseases in developing countries are not known. This lack of knowledge is an obstacle to the adoption of preventive measures which may be of great value in a social and economic environment where treatment options for end-stage renal failure are simply not available to the vast majority of the population. Urinalysis, a simple and inexpensive test, remains a cornerstone in the evaluation of the kidney and may also be easily employed in mass screening for renal abnormalities in a developing country. METHODS: An educational campaign on renal diseases was conducted in three selected areas of Bolivia. Urine samples were collected and sent to one of 21 participating clinical centers. Fresh urine specimens were screened using a dipstick for chemical analysis and by microscopic urinalysis after centrifugation. In those patients in whom urinary abnormalities were found, further investigations were carried out in order to define the diagnosis; these patients were enrolled in a 3-year follow-up program. RESULTS: Apparently healthy subjects (n = 14,082) were referred to the First Clinical and Epidemiological Program of Renal Diseases from rural and metropolitan areas in Bolivia. Urinary abnormalities were detected in 4261 subjects at first screening. The most common form of urinary abnormality was hematuria, which was found in 2010 (47% of positively screened subjects). Other renal abnormalities were leukocyturia (41%) and proteinuria (11%). Confirmatory tests and further clinical studies were then carried out in 1019 people. On a second screening 35% of the subjects had no urinary abnormalities; in the remaining people the following diagnosis were made: asymptomatic urinary-tract infection (48.4%), isolated benign hematuria (43.9%), chronic renal failure (1.6%), renal tuberculosis (1.6%). Other diagnosis were: renal stones 1.3%, diabetic nephropathy 1% and polycystic kidney diseases 1.9%. CONCLUSIONS: This study helped define for the first time the frequency of asymptomatic renal diseases in Bolivia. It shows that it is possible to screen a large population of patients at relatively low cost, providing the framework for further action that may help in the prevention and timely diagnosis of renal diseases. PMID- 9870462 TI - Infection control of hepatitis C in Dutch dialysis centres. AB - BACKGROUND: In dialysis patients, blood transfusions and long-term dialysis are well-known risk factors for transmission of hepatitis C virus (HCV). Transmission of HCV by transfusions has become extremely rare since the introduction of antibody screening. However, nosocomial transmission of HCV within dialysis units still occurs. We performed a survey of current infection control measures against HCV in Dutch dialysis centres that had participated in a national HCV prevalence study. METHODS: All twenty-seven Dutch dialysis centres where HCV-positive patients had been identified (HCV prevalence 1-8%), participated. With the use of a questionnaire we evaluated screening procedures for resident patients and guest patients, routine hygienic measures in HCV-positive and -negative patients, and cleaning procedures of dialysis equipment. RESULTS: All centres except one screened new patients for HCV antibodies, but the frequency of periodic follow-up screening varied. Most centres requested HCV antibody screening of guest patients in advance, but in daily practice 55% of the centres dialysed guest patients even when HCV antibody status was not available. The majority of centres had not implemented special precautions for patients with unknown HCV antibody status. In most centres the use of protective glasses, masks and aprons depended on the HCV antibody status of the patients. Surprisingly, 85% of the centres allowed their nurses to operate dialysis machines with gloves possibly blood contaminated. All centres sterilized their machines at the end of the day, but only 77% sterilized their machines between all dialysis sessions. Traces of blood were removed with alcohol in 63% of the centres. CONCLUSION: Dutch dialysis centres have not yet implemented an optimal policy for prevention of HCV. Especially, operating dialysis machines with gloves might be a potential source for nosocomial transmission of HCV, not yet covered by the issued guidelines. Because dialysis patients probably have a prolonged serological window phase after a recent HCV infection, it does not suffice to implement a preventive strategy against nosocomial transmission based on the results of HCV antibody screening. Universal, rigorous implementation of adequate infection control measures irrespective of HCV antibody status should be the cornerstone for prevention of nosocomial transmission of HCV and other blood borne pathogens. PMID- 9870464 TI - Leukotrienes and lipoxins: lipoxygenase-derived modulators of leukocyte recruitment and vascular tone in glomerulonephritis. AB - With the gradual elucidation of the cellular and molecular events that underpin the inflammatory process, the pathogenetic complexities of glomerulonephritis are slowly being unravelled. Lipoxygenase-derived eicosanoids play important counter regulatory roles within inflamed glomeruli. Leukotrienes, derived from the 5 lipoxygenase pathway, are potent stimuli for leukocyte infiltration, intrarenal vasoconstriction, and mesangial cell contraction in many forms of experimental glomerulonephritis and probably in human disease. The recruitment of 12- and 15 lipoxygenase pathways, particularly during cell-cell interactions, promotes the formation of lipoxins. The latter compounds antagonize many leukotriene effects, attenuate neutrophil recruitment, and are potential 'braking signals' within the inflammatory cascade that promote resolution of inflammation. The generation and metabolism of leukotrienes and lipoxins is regulated independently, and each family of eicosanoids mediates its biological activities through distinct cell surface receptors and signal transduction pathways. Leukotriene biosynthesis inhibitors and leukotriene receptor antagonists are protective in several experimental models of glomerulonephritis. Initial studies with lipoxins and synthetic lipoxin stable analogues suggest that it may be possible to harness this and other putative anti-inflammatory system for therapeutic gain [3,22,92]. PMID- 9870463 TI - CAPD to improve quality of life in patients with refractory heart failure. PMID- 9870465 TI - Effect of heparin on mesangial cell growth and gene expression of matrix proteins. AB - BACKGROUND: Mesangial cell (MC) proliferation and matrix expansion are characteristics of many glomerulopathies. Heparin has been shown to inhibit MC proliferation in vitro and mitigate cell proliferation, matrix expansion, proteinuria, renal insufficiency, and hypertension in experimental glomerulonephritis and subtotal renal ablation. We examined the effect of standard heparin on MC proliferation and matrix protein expression in vitro which necessarily excludes the confounding influences of haemodynamic, inflammatory, haemostatic, and various other processes that are present in vivo. METHODS: Gene expression and release of fibronectin (FN), collagen IV and laminin by cultured rat MC were tested in the presence and absence of heparin. In addition the effect of transforming growth factor-beta1 (TGF-beta1) on the gene expression of those matrix proteins was assessed. RESULTS: Within a 3-1000 microg/ml concentration range, heparin inhibited gene expression and release of FN by 10% fetal calf serum (FCS)-stimulated MC in a concentration-dependent manner. At concentrations of 300 and 1000 microg/ml, heparin inhibited fibronectin mRNA levels in TGF-beta1 (6 ng/ml) stimulated cells. However, heparin had no effect on gene expression or release of collagen IV or laminin under these conditions. Heparin markedly inhibited 10% FCS-stimulated MC proliferation in a concentration-dependent manner. CONCLUSIONS: Heparin inhibited MC growth and fibronectin production. These effects may, in part, account for the reported beneficial effects of heparin on the course of renal disease in experimental animals. PMID- 9870467 TI - Long-term reduction of renal interstitial hydrostatic pressure after neonatal renin-angiotensin system inhibition in the rat. AB - BACKGROUND: Neonatal inhibition of the renin angiotensin system (RAS) causes a decreased urinary concentrating ability, papillary atrophy, and tubulointerstitial inflammation long term. As a consequence of these morphological changes, we surmised that renal blood flow and renal interstitial hydrostatic pressure (RIHP) may be altered during and shortly after cessation of neonatal angiotensin-converting enzyme (ACE) inhibition, and that tentative changes of these variables would persist long after treatment withdrawal. METHODS: Rats were given daily intraperitoneal injections of the ACE inhibitor, enalapril (10 mg/kg) or saline from days 3 to 23 postpartum, and the relationship between renal perfusion pressure (PP) and RIHP was investigated in 6- and 13-week old anaesthetized rats. RESULTS: Neonatal ACE inhibition did not affect baseline RIHP short term, whereas RIHP was reduced at 13 weeks of age versus controls (11.6+/-1.6 vs 18.5+/-1.0 mmHg, P<0.05). Changes in RIHP correlated positively to changes in renal PP, independent of treatment and age (slope averaged 0.11+/ 0.03). Ongoing ACE inhibition until 6 weeks of age neither affected baseline RIHP nor altered the reactivity to changes in perfusion pressure. Mild renal histopathological abnormalities were present already 3 weeks after cessation of treatment and were aggravated significantly in the 13-week-old rats, showing a complete loss of the papillary parenchyma. CONCLUSION: The reduced baseline RIHP in adult rats seemed to constitute a functional correlate to the major papillary atrophy. However, RIHP responses to changes in renal perfusion pressure was maintained, possibly indicating a compensatory effect of the remaining vasa recta and/or peritubular capillary network. Taken together, lack of neonatal angiotensin II type-1 (AT1) receptor stimulation induces not only irreversible abnormalities of the renal architecture, but causes alteration of intrarenal haemodynamics, such as a reduced RIHP, which may have implications for the regulation of pressure-natriuresis. PMID- 9870468 TI - The inheritance of glomerulosclerosis in mice is controlled by multiple quantitative trait loci. AB - BACKGROUND: Glomerulosclerosis, the common terminal event in chronic glomerular diseases such as diabetic nephropathy or IgA nephropathy, leads to end-stage renal disease. The considerable variation in both the risk of developing glomerulosclerosis and the rate of progression in individual patients suggest a role for genetic factors which have not been identified so far. In this study we sought to examine the mode of inheritance of glomerulosclerosis in mice. METHODS: F1 animals of a mating between glomerulosclerosis-prone ROP-Os/+ male and non sclerotic C3H female mice were backcrossed to the ROP strain. We took advantage of the radiation-induced mutation oligosyndactylism (Os) to identify glomerulosclerosis at the age of 3 months. Kidneys were perfused in situ with PBS/Formalin 10%. The extent of glomerulosclerotic lesions was evaluated on PAS stained paraffin sections using computer-aided morphometry. RESULTS: F1 mice did not show any glomerulosclerosis. In the backcross offspring, we found a wide distribution of glomerular lesions between individual animals, ranging from normal to very severe. We calculated that at least 8-10 loci determine the severity of glomerulosclerosis in mice. CONCLUSIONS: Our data show that glomerulosclerosis is inherited in a recessive fashion involving at least 8-10 loci. PMID- 9870466 TI - Reduced binding of immunoglobulin A (IgA) from patients with primary IgA nephropathy to the myeloid IgA Fc-receptor, CD89. AB - BACKGROUND: Primary IgA nephropathy (IgAN) is associated with elevated levels of circulating IgA and is characterized by deposition of primarily IgA1 in the renal mesangium. It has not yet been clarified which mechanisms govern the deposition of IgA1 in the mesangium. One of the factors which may play a role in trapping of IgA in the mesangial area is the interaction of IgA with specific IgA receptors (Fc alphaR, CD89) on the mesangial cells. METHODS: In the present study IgA derived from patients with IgAN and controls was investigated for its interaction with human CD89, expressed on the surface of the murine B cell line IIA1.6. RESULTS: IgA binding to CD89 expressing cells was specific, concentration dependent and binding of dIgA and pIgA occurred in a more efficient fashion than that of mIgA. IgA binding to CD89 directly from serum of patients compared to controls showed no significant difference. However these experiments are affected by differences in IgA concentration and combinations of different sizes of IgA. Using purified fractions of mIgA, dIgA, and pIgA isolated from serum, a significantly reduced binding of mIgA to CD89 from patients compared to controls was observed. Finally, the binding of aIgA2 to CD89 was less inhibited using mIgA from patients with IgAN compared to controls. CONCLUSIONS: The reduced binding of mIgA to CD89 seems to contradict a direct role for CD89 in deposition of IgA. However reduced binding of mIgA to CD89 may affect IgA clearance, leading to higher serum IgA. Furthermore, since it has been demonstrated that mIgA can interfere with binding of di- and pIgA, CD89 could still contribute to pIgA deposition in the mesangial area. PMID- 9870469 TI - Insulin resistance precedes microalbuminuria in patients with insulin-dependent diabetes mellitus. AB - BACKGROUND: Insulin resistance has been associated with hypertension and with renal complications in patients with type 1 diabetes mellitus. Causal relationships have not been fully explained. METHODS: We investigated whether insulin resistance precedes microalbuminuria by measuring insulin resistance with a euglycaemic clamp in combination with indirect calorimetry in 16 uncomplicated type 1 diabetic patients and in six healthy control subjects. The patients had over 10 year duration of diabetes, and were expected to experience either a complication-free or complicated disease course within the next few years. They have thereafter been followed for the development of microalbuminuria for 3 years. RESULTS: In a euglycaemic insulin clamp glucose disposal was lower in diabetic patients compared with control subjects (7.5 +/- 2.9 and 12.6 +/- 2.0 mg/kg LBM/min; P<0.002), mainly due to impaired glucose storage (4.3 +/- 2.3 vs 8.6 +/- 1.6 mg/kg LBM/min; P<0.001). Three years later seven IDDM patients had albumin excretion rate over 30 mg/24 h; glucose disposal (5.5 +/- 2.1 vs 9.0 +/- 2.2 mg/kg LBM/min; P<0.01) had been lower in patients who developed microalbuminuria compared with those who remained normoalbuminuric. CONCLUSIONS: Insulin resistance predicts the increment in urinary albumin excretion. Insulin resistance depends mainly on impaired glucose storage in uncomplicated IDDM. PMID- 9870470 TI - Does treated primary hypertension lead to end-stage renal disease? A 20-year follow-up of the Primary Prevention Study in Goteborg, Sweden. AB - BACKGROUND: Hypertension is reported to be one of the most common causes of end stage renal disease (ESRD) in Europe and in the United States. However, the frequency with which treated primary hypertension leads to renal failure is not known. The majority of patients with ESRD have hypertension. Whether this is the cause or the consequence of the impaired renal function is often not possible to establish. METHODS: To determine if treated primary hypertension can lead to ESRD, we studied the development of serum creatinine levels in 686 white hypertensive men, recruited from a random third of the male population aged 47-55 years living in Goteborg, Sweden (n=9998; 7495 participants). At entry and during 20 years follow-up, all signs of kidney disease, secondary hypertension, or increase in blood pressure were investigated. Records of patients with a serum creatinine value > or = 130 micromol/l at any time during the observation period were thoroughly studied to ascertain the cause of the impaired renal function. RESULTS: A serum creatinine level above 130 micromol was seen in 8.9% (61/686) of the treated hypertensives during the 20 years of follow-up. An underlying renal disorder was found in 7.2% (49/686) of the patients; renoparenchymal disease (2.2%), renovascular disease (1.5%), diabetic nephropathy (1.2%) or a urological disease (1.6%). Only 1.7% (12/686) of the hypertensives showed a moderate progressive increase in serum creatinine of unknown cause. The serum creatinine in this group was 133+/-8 micromol/l (mean+/-SD; range 124-144) after 15 years and 139+/-7 micromol/l (range 132-151) after 20 years. Thus, none of these patients aged 66-71 years had developed ESRD or a clinically important reduction in renal function. CONCLUSION: The main finding in this population-based study of white middle-aged men with primary nonmalignant hypertension was that long-term antihypertensive treatment was not associated with development of end-stage renal disease or even an abnormal progressive decline in kidney function. PMID- 9870471 TI - Audit of antihypertensive treatment in patients with renal failure. AB - BACKGROUND: Recently, intensified antihypertensive treatment has been recommended for patients with proteinuric renal disease, and a target blood pressure of 125/75 mmHg has been proposed. METHODS: In a retrospective crossectional study, all consecutive patients with renal disease and reduced GFR seen in the renal outpatient clinic Heidelberg during a specified time period were analysed. Physicians administered antihypertensive agents according to their clinical judgment aware of recent recommendations, but without structured guidelines. The aims of the study were: (i) to monitor achieved blood pressure (clinic measurement, self measurement, ambulatory BP measurement); (ii) to monitor the number of antihypertensive medications required; and (iii) to analyse the factors which determine achieved blood pressure and the number of antihypertensive agents required. RESULTS: There were 201 non-transplanted patients, median age 60 years (range 20-86), 131 male, 70 female, median S-creatinine 2.33 mg/dl (1.4-10.9). Fifty-two of the patients had diabetes, 41 GN, 18 ADPKD, 17 vasculitis, seven reflux nephropathy, three analgesic nephropathy and 63 other or unknown renal diseases. When the patients had originally been referred to the renal clinic, median clinic BP had been 160 mmHg systolic (104-260) and 95 mmHg diastolic (65 160). Under nephrological care, achieved clinic systolic (140; 90-190) and diastolic BP values (85; 45-130) were significantly (P<0.0001) lower. Achieved BP was slightly (r=0.23, P<0.05) correlated with age, was not significantly different between genders, and was not correlated to proteinuria, serum creatinine, renal disease or BMI. Average self measured BP values were not different from systolic and diastolic clinic BP values. Ambulatory BP values, however, were significantly (P<0.0001) lower, e.g. daytime systolic BP 133 mmHg (107-171), diastolic 82 mmHg (65-97). A night-time decrease of BP > or = 15% was seen in 18/25 patients only. The median number of antihypertensive classes required was 3 (range 1-7) in non-diabetic and 3.5 (1-7) in diabetic patients, respectively. The number of antihypertensive classes was independent of serum creatinine, BMI or proteinuria, but was significantly (P<0.01) higher in males (4; 1-7) than in females (3; 1-6) and correlated to age (r=27, P<0.01). CONCLUSIONS: The study (i) illustrates the difficulty to achieve recommended target BP in patients with renal failure, (ii) shows remarkably little white coat effect on clinic blood pressure, (iii) illustrates the value of ambulatory blood pressure measurement and (iv) documents the importance of multidrug antihypertensive treatment in patients with renal failure. PMID- 9870472 TI - Renal effects of losartan and amlodipine in hypertensive patients with non diabetic nephropathy. AB - BACKGROUND: The objective of this study was to examine the effects of angiotensin II receptor blocker losartan versus the calcium channel blocker amlodipine on proteinuria, renal haemodynamics, glomerular sieving and tubular function in hypertensive patients with non-diabetic nephropathy. METHODS: The study design was a prospective, double blind, placebo controlled, randomized crossover trial with amlodipine and losartan. Renal parameters were measured at baseline and at the end of each 4-week active treatment period. Fifteen patients with a diagnosis of non-diabetic renal disease and hypertension were included. RESULTS: Mean arterial blood pressure decreased from 123+/-13 mmHg at baseline to 113+/-10 mmHg (P<0.01) on losartan and to 114+/-10 mmHg on amlodipine (P<0.01). Urinary albumin excretion significantly decreased from 3510+/-2586 mg/24 h at baseline to 2684+/ 2051 mg/24 h (P<0.01) on losartan and increased non-significantly to 3748+/-3355 mg/24 h on amlodipine. Filtration fraction significantly decreased from a baseline value of 22.8+/-9.3% to 21.2+/-10.2% (P<0.05) on losartan and increased to 23.6+/-8.9% (ns) on amlodipine. Either drug did not significantly alter glomerular sieving of neutral dextrans. CONCLUSION: Our results demonstrate that losartan, but not amlodipine, decreased albumin excretion in hypertensive patients with non-diabetic nephropathy. PMID- 9870473 TI - Clinicopathological features of rapidly progressive hepatitis C virus infection in HCV antibody negative renal transplant recipients. AB - BACKGROUND: Hepatitis C virus (HCV) infection acquired during dialysis treatment generally shows a relatively benign course after renal transplantation (RTx). However, less is known about the course of HCV infection acquired during or after RTx. METHODS: Clinical and histopathological assessment of 15 renal transplant recipients who acquired HCV infection during or after RTx. RESULTS: Alanine aminotransferase levels rose for the first time 1-19 weeks after RTx. HCV RNA was found positive in all patients, but anti-HCV became positive in only nine of them. During a mean follow-up of 21 +/- 12 months, jaundice appeared in 12 patients while ascites and/or hepatic encephalopathy occurred in six. Azathioprine was stopped in all patients. Cyclosporin was also stopped in four patients and in two of them prednisolone was also interrupted for a period of 3-7 weeks. Following this, ascites, hepatic encephalopathy and biochemical disturbances improved, while no deterioration was seen in graft function. Nine of the 15 patients had undergone two consecutive liver biopsies (LB). The first LB revealed cirrhosis in three and chronic hepatitis in six patients; the second LB showed cirrhosis in seven patients. The histological activity index (Knodell's score) progressed from 11.8 +/- 3.5 to 13.8 +/- 3.8. CONCLUSIONS: The results suggest that HCV infection acquired during or after RTx may run an unusual and rapidly progressive clinical and histopathological course at least in some of these patients. Decrease or withdrawal of immunosuppressive drugs may improve early hepatic failure without detrimental effect on graft function during that period. PMID- 9870474 TI - Characterizing flow distributions in AV fistulae for haemodialysis access. AB - BACKGROUND: The arteriovenous fistula is the most widely used means of enabling vascular access for chronic haemodialysis. When interest in vascular access was at its height, in the 1970s, a substantial amount of work was carried out on the physiology of arteriovenous fistulae and on the relationship between fistula flow and patency. One omission in the catalogue of studies was research into the distribution of flow in the fistula and the effects on surgical outcome. METHOD: To address this issue, 30 patients with end-to-side radiocephalic fistulae were studied. Flow measurements were made intraoperatively, and at various intervals postoperatively using colour-flow ultrasonography. RESULTS: In 22 cases, the development of the fistula was monitored at regular intervals for periods of up to 27 months. In seven of the 30 fistulae, flow in the distal artery was antegrade, while in the remaining 23, distal artery flow was retrograde. The proportion of fistula flow supplied by the proximal and distal arteries varied considerably. CONCLUSION: Despite the wide range of flow distributions in the fistulae studied, flow distribution did not appear to affect fistula maturation or long-term function. PMID- 9870475 TI - Assessment of ultrasound guided percutaneous ethanol injection and parathyroidectomy in patients with tertiary hyperparathyroidism. AB - BACKGROUND: Tertiary hyperparathyroidism continues to cause significant morbidity in patients with chronic renal failure. This is frequently resistant to medical management and may ultimately require a surgical parathyroidectomy. Recent studies have reported upon the technique of percutaneous ethanol ablation for both primary and tertiary hyperparathyroidism. In this study we report on a 5 year experience using ethanol injection and compare the results with surgical parathyroidectomy. METHODS: A prospective study in 39 patients with tertiary hyperparathyroidism, 25 were dialysis dependent and 14 had a functioning renal allograft. Twenty-two patients underwent percutaneous fine needle ethanol injection (PFNEI) and 17 underwent surgical parathyroidectomy. RESULTS: A > 30% reduction in intact parathyroid hormone (iPTH) was achieved in 11 of 22 patients undergoing PFNEI after a mean of 1.8 +/- 1.4 injections per gland. In four patients, symptomatic hyperparathyroidism recurred and they required further PFNEI or surgical parathyroidectomy at 17, 28, 46, and 48 months later. There was no significant reduction in iPTH in 11 patients following PFNEI after a mean of 2.5 +/- 1.3 injections per gland. They all required a subsequent surgical parathyroidectomy for symptomatic hyperparathyroidism. Four patients developed a laryngeal nerve palsy following PFNEI, two of which were permanent. Seventeen patients underwent successful surgical parathyroidectomy as a primary procedure. CONCLUSION: Whilst PFNEI is successful in primary hyperparathyroidism, when typically only one adenoma is present, the effectiveness of PFNEI is unpredictable and the long term results are poor compared with those of surgical parathyroidectomy in tertiary hyperparathyroidism. The procedure is not without complications and makes subsequent surgery more difficult. Therefore it can only be recommended for patients with a known single parathyroid gland such as patients in whom hyperparathyroidism has recurred following a previous surgical subtotal parathyroidectomy and who are unsuitable for further surgery. PMID- 9870476 TI - Long-term experience with an ultrapure individual dialysis fluid with a batch type machine. AB - BACKGROUND: This paper discusses long-term experience with a specific type of dialysis equipment which has been used more than 15 years without variation. The system was designed to allow easy individualization of dialysis fluid composition and to deliver dialysate of the highest hygienic quality. METHODS: Data from 399 patients covering the period from 1971 onwards were analysed retrospectively. Survival probabilities were estimated by the Kaplan-Meier method and the median number of days in hospital was calculated. Additional data collected from patient subgroups included serum albumin level, erythropoietin requirement and antihypertensive treatments. Kt/V and PCR from one subgroup were computed using the formulae of Daugirdas and Depner. RESULTS: The estimated survival probability after 5 years for all patients was 59.1% (95% CI: 52.6-65.6%). The main risk factors from the available covariables were age and IDDM. The cumulative incidence of carpal tunnel syndrome after 10 years of dialysis was estimated as 7% (95% CI: 0-14%). Data from the subgroups revealed that 82% of the patients had serum albumin levels >4.0 g/dl, 65% of the patients received no antihypertensive drugs and 39% received erythropoietin (37 +/- 28 units/kg bw/week) to correct dialysis anaemia (haemoglobin level = 98 +/- 8 g/l). Average Kt/V was 1.21 +/- 0.17, PCR was 1.10 +/- 0.22 g/kg/day. CONCLUSIONS: The setup described permits individualized therapy of high quality. The high serum albumin values and our very low incidence of carpal tunnel syndrome underline the importance of water and dialysate quality. PMID- 9870477 TI - Expression of VLA-4 on peripheral mononuclear cells in patients on chronic haemodialysis with carpal tunnel syndrome. AB - BACKGROUND: Carpal tunnel syndrome (CTS) is a major complication in long-term haemodialysis patients and is thought to be a form of haemodialysis-associated amyloidosis. CTS may be due to the deposition of amyloid as well as local inflammatory process in the carpal tunnel. Although macrophage-like cells infiltrating the tenosynovium of carpal tunnel are known to express mRNA for adhesion molecules, the level of expression of adhesion molecules on peripheral blood mononuclear cells (PBMC) in patients with CTS is unknown. METHODS: We compared the expression of very late activation antigen (VLA-4) on PBMC by flow cytometry in 14 patients on haemodialysis (6-21 years) with CTS, nine on haemodialysis (1-6 years) but without CTS and six patients on chronic ambulatory peritoneal dialysis (CAPD, 1-5 years) without CTS. RESULTS: The expression of VLA 4 on peripheral lymphocytes was not different among the groups. However, the FACScan fluorescence histogram of VLA-4 expression on peripheral monocytes showed a bimodal pattern in the CTS group, and the rate of the high intensity portion of the expressed VLA-4 on monocytes was significantly higher in CTS than other groups (P<0.05). The observed differences were not based on differences in the type of the dialysis membrane. We also examined the adhesion of PBMC to fibronectin-coated plates. The number of adherent PBMC was significantly higher in patients with CTS than in other groups (P<0.05). CONCLUSION: Our results suggest that increased expression of VLA-4 on peripheral monocytes and augmented adhesion capacity of PBMC to fibronectin may be involved in the development of CTS in haemodialysis patients. PMID- 9870478 TI - Echinocytosis induced by haemodialysis. AB - BACKGROUND: Haemodialysis, widely used to treat patients with renal failure, is not always well tolerated. Different mechanisms have been postulated for this. We analyzed the influence of haemodialysis on erythrocyte morphology and blood rheology. METHODS: Twenty-two haemodialysed patients were studied immediately before haemodialysis, after 30 min, and at the end of haemodialysis with biocompatible membranes. Haematological routine was measured, the erythrocyte morphology was assessed on glutaraldehyde-fixed cells and blood viscosity was determined. RESULTS: Erythrocytes underwent various degrees of echinocytic shape transformation after 30 min of haemodialysis, which was completely reversible at the end. In a repetition of the investigations during a subsequent haemodialysis other patients were affected. A plasmatic factor caused echinocytosis since the incubation of control erythrocytes in patients plasma induced a similar, even more marked shape transformation and, vice versa, patient echinocytes regained a discocytic shape when incubated in buffer. The degree of echinocytosis was related to an increased blood viscosity at high shear rates (r=0.800, P<0.01). Echinocytosis was not accompanied by obvious clinical reactions. CONCLUSIONS: Reversible echinocytosis and an increase in blood viscosity is often seen during haemodialysis, which may affect the circulation in patients at risk. PMID- 9870479 TI - Urea kinetic modelling--are any of the 'bedside' Kt/V formulae reliable enough? AB - BACKGROUND: Longevity on dialysis is determined by many factors. One of these has increasingly been seen to be 'dialysis dose'. There are several methods for calculating dialysis dose. We wanted prospectively to test 'gold-standard' UKM Kt/V with various shortcut bedside formulae, to see whether reliance on the latter approach was likely to lead to errors in over- or underprescribing dialysis regimens. METHODS: Ten bedside formulae for the calculation of Kt/V (urea) were compared with UKM Kt/V values, in a month-long study involving 507 dialysis sessions in 50 patients in a single-centre in-patient haemodialysis unit. RESULTS: For patients with UKM Kt/V<0.8 (median 0.69, n=140), simplified formulae had a difference (delta) of 0.094-0.396 from the calculated UKM resulting in an inter-method variability ranging from 13 to 57%. The least difference was seen with the Calzavara formula (P=NS), maximum difference with the Barth formulae (P<0.05). No statistically significant differences were seen when comparing Daugirdas 1 and 2 and Keshaviah formulae with UKM, for patients with UKM Kt/V<0.8. For patients with UKM Kt/V in the range 0.8-1.4 (median 1.06, n=285) the extreme recorded values from simplified formulae were 0.012 (least different) and 0.245 (most different) from the UKM mean, with an inter-method variability ranging between 1.1% (Basile method) to 23.1% (Calzavara). No statistically significant difference were seen when comparing Daugirdas 1 and 2, Keshaviah, and Lowrie formulae with UKM, for patients with UKM Kt/V 0.8-1.4. For patients with the highest UKM Kt/V values (>1.4; median 1.58, n=72), all simplified formulae gave Kt/V values lower than UKM Kt/V: the minimum difference was 0.070 using Jindal (P=NS, intermethod variability of 4.4%), while the maximum was seen when using Calzavara (P<0.05; difference = 0.69; intermethod variability of 43.7%). There was also no statistically significant difference for Basile and Kerr methods. For the group as a whole the biggest difference from UKM mean values was obtained using Barth's and Calzavara's formulae (delta of 0.171 and 0.140 respectively (P<0.05)). CONCLUSIONS: The best correlations were seen with the Daugirdas 2 formula (r2=0.953). Also, comparing grouped formulae containing ln(Co/Ct) terms with those incorporating the (Co-Ct)/Co ratio (i.e. the urea reduction) there was a better correlation for all formulae employing the logarithmic transformation (r2=0.951-0.953 cf. r2=0.939-0.940). Nevertheless no bedside formula had the accuracy of UKM-Kt/V. PMID- 9870480 TI - 31P-magnetic resonance spectroscopy (31P-MRS) of human allografts after renal transplantation. AB - BACKGROUND: 31P-Magnetic resonance spectroscopy (31P-MRS) can be used as a non invasive tool for measuring the relative intracellular concentrations of several phosphorus metabolites in different organs. Various pathological conditions are characterized by different metabolic patterns. We studied the value of 31P-MRS after renal transplantation with both an uneventful and a clinically complicated course. METHODS: We determined the relative concentrations of phosphate containing metabolites in renal allografts of humans with 31P-MRS (1.5 Tesla) in the first few weeks after transplantation; 18 patients with an uneventful clinical course and 10 patients who required dialysis after transplantation were examined. Six patients with a stable allograft function 2-3 months after transplantation served as controls. RESULTS: In patients with primary allograft function, we found a significant correlation between the phosphomonoester/phosphodiester-ratio (PME/PDE) (r = 0.66, r < 0.01) and the time after transplantation, but no correlation between the nucleoside triphosphate (beta-NTP)-concentration (r = -0.11) and the time course. In the patients with primary or early allograft dysfunction caused by histologically proven rejection (n=5), we found a low beta-NTP compared to patients with an uncomplicated clinical course (0.09+/-0.01 vs 0.15+/-0.03), but no differences in the PME/PDE ratio (0.73+/-0.21 vs 0.80+/-0.21). In contrast, the PME/PDE ratio was lowered in three patients with delayed graft function caused by acute tubular necrosis (0.45+/-0.07 vs 0.80+/-0.21), but the beta-NTP concentration was not reduced (0.15+/-0.003 vs 0.15+/-0.03). The 31P-MR spectrum of two patients with cyclosporin A damage was not altered compared to the controls. CONCLUSIONS: 31P MRS can be used in patients in the early period after renal transplantation. A significant correlation between the PME/PDE ratio and the time course but no change in the beta-NTP concentration was found in patients with primary allograft function in the first 4 weeks after renal transplantation. Different patterns of 31P-MR spectra were observed depending on the different causes of primary and early transplant dysfunction. PMID- 9870481 TI - Enhancement of human platelet aggregation and secretion induced by rapamycin. AB - BACKGROUND: Rapamycin is a new immunosuppressive drug of the macrolide type. Despite binding to one of the FK-binding proteins as the initial step in intracellular action, further effects differ from those of the other fungally derived macrolides, cyclosporine and tacrolimus. We have previously demonstrated an enhancement of agonist-mediated platelet activation by cyclosporine and tacrolimus which was associated with increased phosphorylation of two intracellular platelet proteins, p20 and p40. Because rapamycin utilizes the same class of binding proteins as tacrolimus, but its action is not associated with the inhibition of calcineurin, we postulated that if the stimulatory effect of cyclosporine or tacrolimus was due to calcineurin inhibition, rapamycin should not affect platelets in a similar fashion. METHODS: Normal, washed human platelets were treated with various concentrations of rapamycin (from ng to microg/ml), and pre-incubated at 37 degrees C with rapamycin for various periods (1-30 min). Several platelet functional parameters were measured in samples treated with rapamycin and these parameters were compared with control platelet samples treated with the vehicle for the same period. Platelet aggregations following exposure to ADP or to the thrombin equivalent, TRAP-6, were measured as changes in optical transmission in a Chronolog lumi-aggregometer. Each experiment was repeated at three or more times and the mean results were used for statistical comparison. RESULTS: Rapamycin-treated platelets demonstrated an increase in their dose- and time-dependent sensitivity to ADP, resulting in a significantly enhanced primary wave of ADP-induced platelet aggregation followed by a secondary wave of aggregation, indicative of granule secretion. Furthermore, rapamycin-treated platelets showed significantly enhanced sensitivity to TRAP-6 as demonstrated by an increase in the initial velocity of aggregation, an increase in their maximal extent of aggregation and an enhancement of granular ATP secretion. Concentrations of rapamycin in the ng range, as well as short pre incubation times (within min), were sufficient to cause significant enhancement of agonist-induced platelet aggregation and secretion (P < 0.001) as compared with their vehicle controls. CONCLUSIONS: Rapamycin significantly potentiates agonist-induced platelet aggregation in a time- and dose-dependent manner. As these findings are similar to those observed with the other fungal macrolides, we hypothesize that inhibition of calcineurin may not be necessary for the increase in intracellular protein phosphorylation observed following exposure of platelets to cyclosporine or tacrolimus. Whether the rapamycin-induced enhancement of sensitivity to agonists and platelet hyperaggregability explains the thrombocytopenia observed in patients when high doses of rapamycin are administered in the clinical setting, and whether these effects are synergistic with cyclosporine, are questions which remain to be investigated. PMID- 9870482 TI - Adherence of Staphylococcus aureus isolated in peritoneal dialysis-related exit site infections to HEp-2 cells and silicone peritoneal catheter materials. AB - BACKGROUND: Peritoneal catheter exit-site infections cause a relevant morbidity in peritoneal dialysis patients and are frequently caused by Staphylococcus aureus. We tested the hypothesis that adherence of exit-site-derived S. aureus to epithelial cells and peritoneal catheter silicone tubes discriminates virulent and less virulent strains. METHODS: The binding of isolated S. aureus to an epithelial cell line (HEp-2) and to silicone tubes was analyzed using light microscopy or radioactive labeling of bacteria. RESULTS: Of 378 exit-site swabs, 99 (26%) were positive for microbial growth. S. aureus was cultured in 25 of 99 positive swabs; three of 13 swabs taken in exit-site infections grade 3 and 4 that had tested positive for S. aureus. Adherence of S. aureus from exit-site infections grade 2, 3 and 4 to Hep-2 cells did not differ from adherence of bacteria isolated from asymptomatic or moderately inflamed catheter exit sites (grade 0-2). However, binding of S. aureus to silicone tubes was enhanced in grade 0/1 compared with grade 2-4 exit-site isolates. CONCLUSIONS: Staphylococcus aureus is an important pathogen in CAPD-related exit-site infection being isolated in about 6.6% of all exit-site swabs (and in 25% of all positive swabs). Silicone-adhesive strains may be of more clinical significance in peritoneal dialysis patients since adhesion to silicone was increased in S. aureus strains isolated in more severe exit-site infections. PMID- 9870484 TI - Sonographic determination of the thickness of the peritoneum in healthy children and paediatric patients on CAPD. AB - BACKGROUND: Prolonged peritoneal dialysis and frequent episodes of peritonitis lead to structural changes and thickening of the peritoneum. Ultrasonography investigations may provide the opportunity to detect morphological changes early, but no systematic investigations have been performed yet. METHODS: Normal values of peritoneal thickness were obtained by systematically examining 131 healthy children (0-15 years) by ultrasound. Parietal peritoneal thickness was best measured at the sternal-umbical line distal from the xiphoid. Growth charts with 95% intervals were prepared. The data of 26 patients with end-stage renal failure (5-18 years) were compared to those of the normal children. RESULTS: The variation coefficient for the consecutive measurements was only 5%, interobserver error was approximately 7%. Whereas gender did not have any influence, peritoneal thickness was significantly correlated to age, weight and most obviously to height (r = 0.81; P<0.001). Children treated only by haemodialysis had normal values, while an increased thickness, loss of movement, and adhesion of the two peritoneal layers were found in children on CAPD. These changes were only noted in patients who had a history of peritonitis. CONCLUSION: Ultrasound examination is a simple, noninvasive and precise method to measure the peritoneal changes in children on CAPD. PMID- 9870483 TI - A controlled trial of two bicarbonate-containing dialysis fluids for CAPD--final report. AB - BACKGROUND: The combination of a low pH and a high concentration of lactate which is present in most dialysis fluids is found to be cytotoxic in vitro. For these reasons it would seem logical to use a bicarbonate-containing solution and thus automatically provide a solution with a neutral pH. METHODS: A parallel, randomized, open-label, prospective 2-month trial with an optional 4 month extension was undertaken to compare two novel bicarbonate-based solutions; one containing 38 mmol/l of bicarbonate (B), and one containing a mixture of 25 mmol/l bicarbonate and 15 mmol/l of lactate (B/L), with a control solution (C) containing 40 mmol/l lactate. RESULTS: Three groups of 19 (C), 20 (B), and 20 (B/L) patients were recruited and data from approximately 55 patient months were accumulated in each group. The data show that both bicarbonate-based solutions maintain acid-base levels within the normal range, that there were no changes in any of the other blood biochemistry parameters measured in the peritoneal equilibration test or with regard to adequacy of dialysis, and that furthermore, both solutions were well tolerated. CONCLUSIONS: This study showed that either the bicarbonate or bicarbonate/lactate solutions could be utilized efficaciously in patients undergoing CAPD. PMID- 9870485 TI - Peritonitis in peritoneal dialysis patients after renal transplantation. AB - BACKGROUND: The occurrence of peritonitis in peritoneal dialysis patients after renal transplantation during immunosuppression might increase morbidity and mortality. Hence the timing of catheter removal is still controversial. The associated risk factors of this complication have not been analyzed. METHODS: We analyzed, retrospectively, the incidence of peritonitis within 90 days after transplantation, its associated morbidity and mortality, as well as risk factors. From 1980 until March 1995, 238 consecutive kidney transplants in peritoneal dialysis patients were performed. Univariate and multivariated logistic regression analysis were used to identify risk factors for the development of peritonitis. RESULTS: 232 cases (141 men, 91 women) were available for analysis. In 191 patients, the catheter was removed with a mean interval after transplantation of 122 days (range 0-573). Thirty peritonitis episodes with predominantly Staphylococcus aureus (10/30) or gram-negative bacteria (12/30) were observed. Independent risk factors before transplantation were the total number of peritonitis episodes (P<10(-5)), previous peritonitis with S. aureus bacteria (P<10(-5)), and male sex (P<0.004). Risk factors after transplantation were technical surgical problems (P<10(-5)), more than two rejection episodes (P<0.02), permanent graft nonfunction (P<0.026), and urinary leakage (P<0.035). CONCLUSIONS: Transplantation without simultaneous peritoneal catheter removal is feasible. However, this increases the risk of peritonitis after transplantation. Early catheter removal should be considered seriously in those patients at risk. When peritonitis develops, antibiotic treatment should be directed against gram positive as well as gram-negative bacteria until culture results are available. PMID- 9870486 TI - Polymorphisms in angiotensin-converting enzyme gene and severity of renal disease in Henoch-Schoenlein patients. Italian Group of Renal Immunopathology. AB - BACKGROUND: The influence of angiotensin converting enzyme (ACE) gene polymorphism on the progression of primary IgA nephropathy (pIgAN) is still debated. Even though the allele frequency was reported to be similar to controls, in some studies D/D patients had a faster decline of renal function and need of dialysis. Since Henoch-Schoenlein purpura (HSP) nephritis is considered a systemic vasculitis with renal lesions indistinguishable from pIgAN, we investigated the effect of the ACE polymorphism on presentation and progression of HSP IgAN. METHODS: We examined the insertion (I) and deletion (D) polymorphism in intron 16 of ACE gene by PCR amplification of genomic DNA of 82 patients (37 children), with biopsy-proven IgAN associated with HSP enrolled in a collaborative study. RESULTS: No significant association with clinical presentation at onset or with final outcome was found (functional impairment at outcome in 31.8%, D/D, 27.4%, I/D and 44% I/I, heavy proteinuria in 36.3% D/D, 21.6% I/D, and 11.1% I/I). Patients homozygous for the D allele had a greater number of extrarenal relapses (P=0.0028). No association was found between the ACE genotype and the presence of hypertension at onset and at the end of the follow-up. No difference was found between adults and children. CONCLUSIONS: In this cohort of HSP IgAN, no ACE I/D polymorphisms were found to be associated with progressive deterioration of renal function. Different genes possibly involved in vasculitis might more strictly modulate expression and evolution of HSP IgAN. PMID- 9870487 TI - Automated peritoneal dialysis with 'on-line'-prepared bicarbonate-buffered dialysate: technique and first clinical experiences. AB - BACKGROUND: Automated peritoneal dialysis (APD) has the possibility of increasing the dialysis efficacy by using higher fill volumes, frequent dialysate exchanges, and tidal techniques. It is then possible to treat patients adequately without residual renal function. The drawbacks of the required high amounts of dialysis solution of up to 30 litres per session are the high costs of lactate-based dialysate bags and difficulties for the patients in handling these bags. So far, bicarbonate-based peritoneal dialysate, which may be more biocompatible, is only available for CAPD in double-chamber bags. In APD this could be overcome by 'on line' preparation of bicarbonate-buffered dialysate using advanced technologies originally designed for on-line preparation of substitution fluid for haemofiltration. METHODS: Four patients without residual renal function were treated with APD five times weekly in a crossover study design. Patients received standard lactate-based (35 mmol/l) treatment (25 litres per session each) in weeks 1 and 3. In week 2 on-line-produced bicarbonate-buffered (37 mmol/l) dialysate was used. This dialysate was prepared by an AK 100 Ultra haemodialysis machine. The machine was modified for adding glucose from a 50% concentrate to the desired concentration of 1.7%. Electrolytes, pH, pCO2, and dialysis efficacy parameters were measured. Microbiological testing was carefully performed. RESULTS: Creatinine clearances, Kt/V, and pCO2 did not vary between the different treatment phases, whereas the pH showed a distinct increase during the bicarbonate phase. Repeated determinations of endotoxins and culturing showed no contamination of the dialysate. The composition of the produced dialysate was reproducible with respect to pH, pCO2, sodium, calcium and bicarbonate, whereas the glucose concentration varied by +/- 20%. CONCLUSIONS: On-line preparation of PD fluid with the AK 100 Ultra is easy and safe to handle. APD with dialysate containing 37 mmol/l bicarbonate provides improved acid base balance and possibly improved biocompatibility, and may lead to a significant cost reduction. Further development in order to provide smaller machines and more precise ways of achieving a desired dialysate glucose concentration is necessary. PMID- 9870488 TI - Amelogenesis imperfecta, nephrocalcinosis, and hypocalciuria syndrome in two siblings from a large family with consanguineous parents. PMID- 9870489 TI - Haemolytic-uraemic syndrome in a heroin addict. PMID- 9870490 TI - Cutaneous botryomycosis in a patient with lupus nephritis. PMID- 9870491 TI - Papillary muscle rupture complicating inferior myocardial infarction in a young woman with systemic lupus erythematosus and antiphospholipid syndrome. PMID- 9870492 TI - Selective thrombolysis in a newborn with bilateral renal venous and cerebral thrombosis and heterozygous APC resistance. PMID- 9870493 TI - Pneumonia and glomerulonephritis caused by Mycoplasma pneumoniae. PMID- 9870495 TI - Emphysematous pyelonephritis in two diabetic patients with complete uterine prolapse and cystocele. PMID- 9870494 TI - Mycophenolate mofetil in cyclosporin-associated thrombotic microangiopathy. PMID- 9870496 TI - Nephropathy and neuropathy induced by a germanium-containing compound. PMID- 9870497 TI - Recurrent infections in haemodialysis patients--do not forget selective immunoglobulin A deficiency. PMID- 9870498 TI - Early destructive spondyloarthropathy from combined beta2-microglobulin and transthyretin Met30 amyloidosis in a dialysed patient. PMID- 9870499 TI - Heparin-induced thrombocytopenia (HIT) and thrombosis in a haemodialysis dependent patient with systemic vasculitis. PMID- 9870500 TI - Aortic regurgitation caused by complex outflow calcification in a patient with chronic renal failure. PMID- 9870501 TI - Penile necrosis in a haemodialysis patient: a rare manifestation of cholesterol crystal embolism. PMID- 9870502 TI - Local recurrence of renal cell carcinoma and acquired cysts 10 years after tumour nephrectomy. PMID- 9870504 TI - Diffuse T-cell lymphoma in a kidney graft recipient 17 years after transplantation. PMID- 9870503 TI - Transitional-cell carcinoma in a 25-year-old renal allograft. PMID- 9870505 TI - Low-back pain caused by spinal tophus--a complication of gout in a kidney transplant recipient. PMID- 9870506 TI - Positive captopril renography in a transplanted kidney with arteriovenous fistula. PMID- 9870507 TI - Renal transplantation for end-stage renal disease due to paroxysmal nocturnal haemoglobinuria. PMID- 9870508 TI - Neurovascular decompression of the left ventrolateral medulla as a treatment of hypertension in a patient with renal artery stenosis. PMID- 9870509 TI - Pulmonary mucormycosis presenting as fatal massive haemoptysis in a renal transplant recipient. PMID- 9870510 TI - The patient developing Wallenberg's syndrome during plasma separation. PMID- 9870511 TI - A winking warning. PMID- 9870512 TI - LE cell in tissue biopsy. PMID- 9870513 TI - Pregnancy during dialysis: case report and management guidelines. PMID- 9870514 TI - Professor Mollie McGeown. PMID- 9870515 TI - Minimal change nephrosis and antiphospholipid antibodies: coincidental or associated? PMID- 9870516 TI - Response of mean reticulocyte haemoglobin content to intravenous iron therapy in haemodialysis patients. PMID- 9870517 TI - Presentation of tumoral calcinosis with systemic signs pointing to inflammatory disease. PMID- 9870518 TI - Kaposi's sarcoma after renal transplantation--disappearance after reduction of immunosuppression and reappearance 7 years later after start of mycophenolate mofetil treatment. PMID- 9870519 TI - Reactivation of multisystemic sarcoidosis after immunosuppression withdrawal in a transplanted patient returning to chronic dialysis. PMID- 9870521 TI - Alpha-lactalbumin as a modulator of mammary cellular activity. PMID- 9870520 TI - Immuno-epithelial interactions: cytokine modulation of normal rabbit colonocyte function. PMID- 9870522 TI - Cyclophosphamide-induced chromosomal aberrations and associated congenital malformations in rats. PMID- 9870523 TI - Further steps in the initiation of cell cultures from embryos and adult sponge colonies. PMID- 9870524 TI - Development of a serum-free medium for the production of humanized antibody from Chinese hamster ovary cells using a statistical design. AB - To develop serum-free (SF) media for the production of humanized antibody from recombinant Chinese hamster ovary (rCHO) cells, a statistical optimization approach based on a Plackett-Burman design was adopted. A basal medium was prepared by supplementing alpha-minimal essential medium (alpha-MEM) with Fe(NO3)3.9H2O, CuCl2, ZnSO4.7H2O, and Na2SeO3 which are generally contained in SF medium formulations. Insulin, transferrin, and ethanolamine were also supplemented to the basal medium to determine their optimal concentrations. From this statistical analysis, serine, phenylalanine, and tyrosine were identified as important determinants for cell growth. Also, putrescine, linoleic acid, and hydrocortisone were shown to be important for both cell growth and antibody production. The SF medium was formulated by supplementing the basal medium with components showing positive effects on cell growth and/or antibody production. Cell growth and antibody production in this SF medium were comparable to those in alpha-MEM supplemented with 5% dialyzed fetal bovine serum. Taken together, the results obtained here show that a Plackett-Burman design facilitates the development of SF media for rCHO cells aimed at producing a humanized antibody. PMID- 9870525 TI - Viability and differential function of rainbow trout liver cells in primary culture: coculture with two permanent fish cells. AB - The study investigates the influence of different culture conditions on attachment, viability and functional status of rainbow trout (Oncorhynchus mykiss) liver cells in primary culture. Cells were isolated by a two-step collagenase perfusion and incubated in serum-free, chemically defined minimal essential medium (MEM), (a) as a monolayer on uncoated PRIMARIA dishes, (b) as a monolayer on culture dishes coated with calf collagen type 1, and (c) in coculture with the established fish cell lines RTH-149 or RTG-2. Cell attachment was assessed from DNA and protein concentrations per dish, viability was estimated from cellular lactate dehydrogenase release, and the metabolic status was investigated by measuring activities of the phosphoenolpyruvate carboxykinase and biotransformation enzymes as well as the total cytochrome P450 contents. Seeding of hepatocytes on collagen-coated dishes did not alter cell attachment or detachment from the (culture substrate, but had a small, but not significant effect on cell viability and metabolic parameters. Coculture of liver cells and RTG-2 cells reduced hepatocyte detachment from the culture substrate, and it was associated with a significant elevation of 7-ethoxyresorufin-O-deethylase activities in the hepatic cells. Cytochrome P450 contents, however, were not altered. The coculture effect on liver cell physiology clearly depended on the type of cell line, because coculture with RTH-149 cells led to similar, but much weaker effects than obtained in cocultures with RTG-2 cells. Electron microscopical observations revealed the existence of gap junctions and possible exocytosis-like transport between cell lines and hepatocytes. The results point to the potential of coculture systems to improve physiological parameters of trout liver cells in primary culture. PMID- 9870526 TI - A fluorometric assay for the measurement of endothelial cell density in vitro. AB - A fluorometric assay for determining endothelial cell numbers based on the endogenous enzyme acid phosphatase is described. In preliminary studies, three substrates--p-nitrophenyl phosphate, 4-methylumbelliferyl phosphate, and 2'-[2 benzthiazoyl]-6'-hydroxy-benthiazole phosphate (AttoPhos)--were compared with respect to their kinetic, optimum assay conditions, sensitivity, and detection limits. Only AttoPhos was found to have a high degree of sensitivity, reliability, and reproducibility for measuring both high and low cell numbers in the same plate. In subsequent experiments, assay conditions were validated for measuring endothelial cell density in response to basic fibroblast growth factor and fumagillin. Furthermore, the AttoPhos assay revealed a linear correlation between acid phosphatase activity and cell number in many cell types, including BALB/3T3, CHO-K1, A431, MCF7, 2008, SK-OV-3, T47-D, and OVCAR-3. This assay is potentially valuable for use in many in vitro systems in which the quantitation of cell density and proliferation is necessary. The practical advantages of AttoPhos assay for measuring endothelial cell numbers include (1) nonradioactivity, (2) simplicity, (3) economy, (4) speed of assessment of proliferation of large number of samples, and (5) amenability to high-throughput drug screening. PMID- 9870527 TI - Immortalized mouse brain endothelial cells are ultrastructurally similar to endothelial cells and respond to astrocyte-conditioned medium. AB - Studies of brain microvessel endothelial cell physiology and blood-brain barrier properties are often hampered by the requirement of repeatedly producing and characterizing primary endothelial cell cultures. The use of viral oncogenes to produce several immortalized brain microvessel cell lines has been reported. The resulting cell lines express many properties of the blood-brain barrier phenotype but do not completely mimic primary endothelial cells in culture. As immortalized brain microvessel endothelial cell lines have not yet been produced from mice, we transformed mouse brain endothelial cells with the adenovirus E1A gene using a retroviral vector (DOL). Eight of 11 clones produced exhibited an endothelial like cobblestone morphology and were characterized as endothelial with a panel of antibodies, lectins, and ultrastructural criteria. These cells are endothelial in origin and share ultrastructural features with primary cultures of endothelial cells. Examination of freeze fracture and transmission electron micrographs show adherens junctions exist between the transformed cells, and culture in astrocyte conditioned medium induces the formation of gap junctions. This is one indication that responses to astrocyte-derived factors are retained by the transformed cell lines. PMID- 9870529 TI - Morphological and molecular characterization of new Drosophila cell lines established from a strain permissive for gypsy transposition. AB - The gypsy element of Drosophila melanogaster is the first retrovirus identified in invertebrates. Its transposition is controlled by a host gene called flamenco (flam): restrictive alleles of this gene maintain the retrovirus in a repressed state while permissive alleles allow high levels of transposition. To develop a cell system to study the gypsy element, we established four independent cell lines derived from the Drosophila strain SS, which contains a permissive allele of flamenco, and which is devoid of transposing copies of gypsy. The ultrastructural analysis of three SS cell lines revealed some remarkable characteristics, such as many nuclear virus-like particles, cytoplasmic dense particles, and massive cisternae filled with a fibrous material of unknown origin. Gypsy intragenomic distribution has been compared between the three cell lines and the original SS fly strain, and revealed in two of the cell lines an increase in copy number of a restriction fragment usually present in active gypsy elements. This multiplication seems to have occurred during the passage to the cell culture. Availability of SS cell lines should assist studies of gypsy transposition and infectivity and might be useful to produce high amounts of gypsy viral particles. These new lines already allowed us to show that the Envelope-like products of gypsy can be expressed as membrane proteins. PMID- 9870528 TI - Selective and organotypic culture of intrahepatic bile duct cells from adult pig liver. AB - Secondary culture of nontransformed bile duct epithelium has been difficult to achieve. STO feeder cell-dependent secondary cultures of adult pig bile duct cells were established from primary cultures of adult pig liver cells. Adult pig hepatocytes exhibited limited or no replication and were lost from the secondary culture at Passage 3 or 4. In contrast, adult pig bile duct cells replicated and were carried for 4-8 passages in secondary culture. A simple method to produce nearly pure pig intrahepatic bile duct cultures was first to freeze a relatively crude liver cell preparation. Upon subsequent thawing, all hepatocytes and most macrophages were lysed. Bile duct cells composed 95% of the surviving cells after the freeze/thaw, and they grew out rapidly. The bile duct cells grew on top of the STO feeder cells as closely knit epithelial, colonial outgrowths. Histocytochemical and biochemical analyses demonstrated high levels of gamma glutamyltranspeptidase activity and low levels of P450 activity in the bile duct cultures. The bile duct cells spontaneously adopted a multicellular ductal morphology after 7-10 d in static culture which was similar to that found in in vivo pig liver. Transmission electron microscopic examination revealed complex junctions and desmosomes typical of epithelium, and lumenally projecting cilia typical of in vivo intrahepatic bile ductules. This simple method for the coculture of pig intrahepatic bile duct cells which adopt in vivo-like structure may facilitate biological studies of this important, but difficult to culture, cell type. PMID- 9870530 TI - Enhanced neointimal growth in cultured rabbit aorta following in vivo balloon angioplasty. AB - We have used in vivo balloon catheterization in combination with in vitro organ culture to develop a model system for vascular neointima formation. A Fogarty balloon catheter was used to deendothelialize and rupture the internal elastic lamina of aortae in adult rabbits. After three d of recovery, aortae were harvested, divided into segments, and placed into organ culture. We obtained a daily index of cell proliferation in cultured vessels using [3H]thymidine incorporation into DNA. Also, segments were collected and processed for routine histology or immunohistochemistry. Aortic segments that had undergone ballooning 3 d before harvest and then cultured exhibited diffuse neointimal growth after several d in vitro, whereas those from sham-operated (nonballooned) rabbits showed generally only a single endothelial cell layer that is characteristic of normal intima. Aortae that were harvested, balloon-damaged in vitro, and then cultured exhibited no neointimal growth. The neointima that developed in cultured segments from in vivo ballooned rabbits was primarily of smooth muscle cell origin as determined by positive immunostaining for alpha-smooth muscle actin. The intima:media thickness ratios were significantly higher in aortic segments from ballooned rabbits at harvest and after 4 or 7 d in culture compared with those from nonballooned rabbits. Also, the [3H]thymidine index was higher in the in vivo ballooned aorta compared to non-ballooned or in vitro ballooned vessel. We conclude that ballooning in vivo followed by exposure to blood-borne elements produces an enhanced proliferative response in cultured vessels that is distinct from other in vitro models of neointimal growth. PMID- 9870531 TI - Ultrastructural changes of Drosophila larval and prepupal salivary glands cultured in vitro with ecdysone. AB - Alterations in the ultrastructure of in vitro cultured larval salivary glands of Drosophila melanogaster in response to the steroid hormone ecdysone were studied in relation to complex changes in puffing patterns. We found that the changes in the fine structure of cultured glands reflected progression of the puffing pattern, and they paralleled those seen in vivo. We observed that glue secretion by exocytosis, the main function of salivary glands, took place between puff stage 5 (PS5) and PS7. Glue could not be expectorated under culture conditions but was slowly released from the lumen through a duct into the medium. After the cultured glands reached PS13/PS14, further progress of puffing and fine structural alterations required that the ecdysteroid titer be transiently extremely low or absent. Under in vitro conditions we did not observe the putative new secretory program(s) described for glands in vivo after PS12. However, ultrastructural changes which unambiguously indicated that an autohistolytic process had begun in vitro started to appear after PS17. Many salivary gland cells developed numerous features of progressive self-degradation between PS18 and PS21. Actual degradation of salivary glands in vivo seemed to be rapid, but in vitro degradation was never completed, probably due to a lack of exogenous factors from the hemolymph. Manipulations of ecdysone titer in vitro in the culture medium, known during the larval puffing cycle to cause premature induction of developmentally specific puffing patterns, did not affect the normal development of ultrastructural features of the cytoplasm and nucleus. PMID- 9870532 TI - Epidermal growth factor accelerates recovery of LLC-PK1 cells following oxidant injury. AB - In renal tubular epithelial cells, oxidant injury results in several metabolic alterations including ATP depletion, decreased Na+K+ATPase activity, and altered intracellular sodium and potassium content. To investigate the recovery of LLC PK1 cells following oxidant injury and to determine if recovery can be accelerated, we induced oxidant stress in LLC-PK1 cells with 500 microM hydrogen peroxide for 60 min. Identical cohorts of oxidant-stressed cells were incubated in recovery medium without epidermal growth factor (EGF) or recovery medium containing 25 ng EGF per ml. ATP levels, Na+K+ATPase activity in whole cells, Na+K+ATPase activity in disrupted cells, and intracellular sodium and potassium ion content were determined at 0, 5, 24, 48, and 72 h following oxidant injury in each cohort of cells. In oxidant-stressed cells recovering in medium without EGF, ATP levels, Na+K+ATPase activity, and intracellular ion content improved but continued to remain substantially lower than control values at all time points following oxidant stress. In cells recovering in medium with EGF, ATP levels, Na+K+ATPase activity, and the intracellular potassium-to-sodium ratio were significantly higher at nearly all time points than values in cells recovering in medium alone. In cells recovering with added EGF, Na+K+ATPase activity had improved to control levels, whereas ATP levels and intracellular ion content approached control values by 72 h following oxidant stress. We conclude that oxidant-mediated ATP depletion, altered Na+K+ATPase activity, and intracellular ion content remain depressed for several d following oxidant stress and that EGF accelerated recovery of LLC-PK1 cells from oxidant injury. PMID- 9870533 TI - MSX-1 gene expression and regulation in embryonic palatal tissue. AB - The palatal cleft seen in Msx-1 knock-out mice suggests a role for this gene in normal palate development. The cleft is presumed secondary to tooth and jaw malformations, since in situ hybridization suggests that Msx-1 mRNA is not highly expressed in developing palatal tissue. In this study we demonstrate, by Northern blot analysis, the expression of Msx-1, but not Msx-2, in the developing palate and in primary cultures of murine embryonic palate mesenchymal cells. Furthermore, we propose a role for Msx-1 in retinoic acid-induced cleft palate, since retinoic acid inhibits Msx-1 mRNA expression in palate mesenchymal cells. We also demonstrate that transforming growth factor beta inhibits Msx-1 mRNA expression in palate mesenchymal cells, with retinoic acid and transforming growth factor beta acting synergistically when added simultaneously to these cells. These data suggest a mechanistic interaction between retinoic acid, transforming growth factor beta, and Msx-1 in the etiology of retinoic acid induced cleft palate. PMID- 9870535 TI - Caries prediction: a review of the literature. AB - For over a decade researchers have been looking for the formula that will allow caries prediction. The purpose of this summary is to review recent multifactorial prediction models for adults and children with the aim of identifying the most successful and consistent methods. Conclusions from this review include the following: Clinical variables, especially past caries experience, are confirmed as the most significant predictors of future caries development. The status of the most recently erupted/exposed surface is the most successful measure of past caries experience. Bacterial levels are included in the most accurate prediction models. Sociodemographic variables are most important to caries prediction models for young children and older adults. PMID- 9870534 TI - Responsiveness to hormone, growth factor and drug treatment of a human breast cancer cell line: comparison between early and late cultures. AB - Growth rate, morphology, and responsiveness to mitogenic stimuli and pharmacological treatments were evaluated in early and late cell passages derived from the same clone of the widely used MCF-7 human breast adenocarcinoma cell line. Our results indicate dissimilarities between early (E) and late (L) passages for some of the parameters analyzed. The cells that underwent many subcultivations grew faster than the others; both appeared homogeneous in size and shape. The E cells, subcultured for almost 1 yr, displayed higher sensitivity to the mitogenic action of both estradiol, according to the level of estrogen receptor, and insulin-like growth factor-I than did the L cells, kept in culture for more than 10 yr. Cell responsiveness to two drugs, a novel steroid antiestrogen and a polysulfonated distamycin A derivative, was more pronounced in the early cultures only at the longer time of exposure to the higher concentration of the estrogen antagonist. In addition, a drug-induced inhibition of insulin-like growth factor-I binding to its receptor was shown in both E and L cells, the latter being less sensitive than the former when exposed to the antiestrogen. Finally, MCF-7 E and L cells showed similar behavior when drug induced apoptosis was tested. PMID- 9870536 TI - ART restorations and glass ionomer sealants in Zimbabwe: survival after 3 years. AB - Atraumatic restorative treatment (ART) consists of removing demineralised tooth tissues with hand instruments only, restoring the prepared cavity and sealing the adjacent pits and fissures with an adhesive filling material. This relatively painless, no-handpiece, minimal intervention approach to controlling dental caries is described. ART was applied in an oral health care programme in Zimbabwe that was carried out amongst secondary school students from 1994 to 1997. A new glass ionomer (Fuji IX) was used as the restorative and sealant material. Sealants were placed in high caries risk students using the 'press-finger' technique. A total of 297 one-surface ART restorations and 95 glass ionomer sealants were placed in 142 and 66 students, respectively. After 3 years, the lost-to-follow-up percentages for one-surface ART restorations and glass ionomer sealants were 30.6% and 30.5%, respectively. Actuarial (life table) analysis resulted in 3-year survival rates of one-surface ART restorations of 88.3% (95% CI: 92.4%-84.2%), ranging from 94.3% to 65.4% per operator. A total of 28 ART restorations placed in 25 students failed. Reasons for failure related to the material and the operator (11 restorations or 5.3% each), and to caries adjacent to the restoration (one restoration or 0.5%). Reasons for failure were not recorded for five restorations (2.3%). Sealants were placed on surfaces diagnosed as early enamel lesions. After 3 years, 71.4% (95% CI: 81.7%-61.1%) of the fully and partially retained sealants survived with a range of 100% to 55.6% per operator. Of the sealed surfaces 96.3% (95% CI: 100%-92.2%) survived 3 years without developing caries. Experienced operators placed better ART restorations than inexperienced operators. This study has demonstrated that ART with a glass ionomer restorative material and sealants provided high quality preventive and restorative dental care to this student population. ART has become one of the treatment modalities available to oral health workers in managing dental caries. PMID- 9870537 TI - Severe dental fluorosis in a Tanzanian population consuming water with negligible fluoride concentration. AB - OBJECTIVES: To identify risk factors for dental fluorosis that cannot be explained by drinking water fluoride concentration alone. METHODS: Two hundred eighty-four Tanzanian children ages 9 to 19 (mean 14.0+/-SD 1.69), who were lifetime residents at differing altitudes (Chanika, 100 m; Rundugai, 840 m; and Kibosho, 1,463 m; Sites 1, 2, and 3 respectively) were examined for dental fluorosis and caries. They were interviewed about their food habits, environmental characteristics and use of a fluoride-containing food tenderizer known locally as magadi. Meal, urine, water and magadi samples supplied by the participants were analyzed for fluoride content. Urine samples were also analyzed for creatinine concentration. Four magadi samples from Sites 1 and 3 were analyzed for complete element composition. RESULTS: Of the 13 water samples from Site 2, 10 contained > or =4 mg/L F, ranging from 1.26 to 12.36 mg/L with a mean+/-SD of 5.72+/-4.71 mg/L. Sites 1 and 3 had negligible water fluoride of 0.05+/-0.05 and 0.18+/-0.32 mg/L respectively. Mean TFI fluorosis scores (range 0 9) for Site 2 were high: 4.44+/-1.68. In Sites 1 and 3, which both had negligible water fluoride, fluorosis scores varied dramatically: Site 1 mean maximum TFI was 0.01+/-0.07 and Site 3 TFI was 4.39+/-1.52. Mean DMFS was 1.39+/-2.45, 0.15+/ 0.73 and 0.19+/-0.61 at Sites 1, 2, and 3, respectively. There were no restorations present. Urinary fluoride values were 0.52+/-0.70, 4.34+/-7.62, and 1.43+/-1.80 mg/L F at Sites 1, 2, and 3, respectively. Mean urinary fluoride values at Site 3 were within the normal urinary fluoride reference value range in spite of pervasive severe pitting fluorosis. Meal and magadi analyses revealed widely varied fluoride concentrations. Concentrations ranged from 0.01 to 22.04 mg/L F for meals and from 189 to 83211 mg/L F for magadi. Complete element analysis revealed the presence of aluminum, iron, magnesium, manganese, strontium and titanium in four magadi samples. There were much higher concentrations of these elements in samples from Site 3, which was at the highest altitude and had severe enamel disturbances in spite of negligible water fluoride concentration. An analysis of covariance model supported the research hypothesis that the three communities differed significantly in mean fluorosis scores (P<0.0001). Controlling for urinary fluoride concentration and urinary fluoride:urinary creatinine ratio, location appeared to significantly affect fluorosis severity. Urinary fluoride:urinary creatinine ratio had a stronger correlation than urinary fluoride concentration with mean TFI fluorosis scores (r=0.43 vs r= 0.25). CONCLUSIONS: The severity of enamel disturbances at Site 3 (1463 m) was not consistent with the low fluoride concentration in drinking water, and was more severe than would be expected from the subjects' normal urinary fluoride values. Location, fluoride in magadi, other elements found in magadi, and malnutrition are variables which may be contributing to the severity of dental enamel disturbances occurring in Site 3. Altitude was a variable which differentiated the locations. PMID- 9870538 TI - Compliance in dentistry: general adherence, specific adherence and perceived dental health. AB - In this study it was attempted to replicate findings on the relationship between general and specific adherence and perceived health in the context of dental treatment. Furthermore the psychometric characteristics of a modified adherence scale for use in dental populations were investigated. A sample of 167 freshmen completed a questionnaire containing a general adherence scale (GAS) developed by DiMatteo et al. (Oncology 1992;6:50-7), a perceived health scale, a perceived dental health scale, and two items about adherence to recommendations on dental behaviour. The GAS showed adequate internal consistency. Positive correlations were found for general adherence on the one hand and for specific adherence, perceived health, and perceived dental health on the other. The results concerning the GAS and specific adherence do not correspond with the findings of DiMatteo et al. It is suggested that this discrepancy is due to the situation specificity of adherence in combination with differences in the samples and the specific adherence measures used in both studies. PMID- 9870539 TI - Measuring patient satisfaction with mandibular prostheses. AB - OBJECTIVES: Previous research has shown that patients' evaluations of their prostheses correlate poorly with the clinicians' assessments, as well as with intraoral anatomic factors. It has been recommended that researchers conduct more studies that use patient satisfaction as the primary outcome measure in treatment evaluation and that more attention be paid to understanding exactly what measures of patient satisfaction represent. In this study, the relationship between patients' ratings of general satisfaction and their perceptions of different aspects of mandibular prostheses is investigated. METHODS: One hundred and twenty subjects applied to participate in a randomized controlled clinical trial comparing two types of mandibular prostheses: conventional dentures and implant prostheses. At baseline, they were asked to rate on 100 mm visual analog scales (VAS) factors that edentulous patients indicated were important to them. These included comfort, ability to chew, stability, esthetics, ability to speak and ease of cleaning of their conventional dentures. Subjects were also asked to rate their general satisfaction with their dentures. In addition, they selected the one quality of their denture that they considered to be most important. RESULTS: Multiple regression methods revealed that gender, as well as patients' ratings of comfort, stability, esthetics, ability to chew and ability to speak with their prostheses contributed significantly to general satisfaction (F<0.0001). Furthermore, 89% of the variation in ratings of general satisfaction was explained by these factors. In addition, patients who considered ability to chew as the most important factor associated with their dentures rated their general satisfaction significantly higher than the other subjects (P=0.0003). CONCLUSION: Patient satisfaction with conventional dental prostheses is highly dependent on gender, and the appearance and functionality of the appliance. The combined effect of these factors explained most of the variation in the satisfaction ratings. PMID- 9870540 TI - The influence of toothbrushing frequency and post-brushing rinsing on caries experience in a caries clinical trial. AB - OBJECTIVE: To examine the effect of reported toothbrushing frequency and method of rinsing after brushing on caries experience and increment. METHODS: Data are presented from 2621 adolescents (mean age 12.5 years at outset) participating in a 3-year double-blind caries clinical trial. At baseline, examiners questioned each participant about their toothbrushing habits, and at subsequent examinations, this information was obtained using a self-administered computer based questionnaire. Participants used a fluoride-containing dentifrice throughout and clinical examinations were conducted using a mirror, CPITN probe and fibre-optic transillumination. RESULTS: The reported brushing frequency increased throughout the trial. Caries experience at baseline was inversely related to toothbrushing frequency with mean DMFS=9.66, 8.12 and 7.63 respectively for <1/day, 1/day and >1/day brushers (P<0.001). Mean 3-year DMFS increments of 8.90, 6.63 and 5.48 (P<0.01) were observed in those reporting to brush <1/day, 1/day or >1/day, on not less than two of the three clinical examinations during the trial. Caries increment was also significantly related to the claimed method used to rinse post-brushing. Overall frequency of brushing and rinsing method accounted for over 50% of the explained variance in the ANOVA model used to analyse the DMFS increments. CONCLUSIONS: Stated toothbrushing frequency and rinsing method after brushing were found to be strongly correlated with caries experience and caries increment. These factors should be reflected in the design of oral health education material and taken into account in the design and analysis of caries clinical trials. PMID- 9870541 TI - Association of salivary mutans streptococci with discoloured pits and fissures. AB - Experimental studies have demonstrated that mutans streptococci play a major role in caries etiology. Several previous epidemiologic studies found a positive association of salivary mutans streptococci with caries prevalence. The present epidemiologic study aimed at detecting a possible association of salivary mutans streptococci with brown discoloured pits and fissures, supposing that discolouration indicates caries. In the Canton of Zurich 1035 schoolchildren, aged 6.5-12.5, were examined with regard to caries prevalence and presence of discolourations in pits and fissures. A commercially available, semi-quantitative test was used to estimate the salivary level of mutans streptococci in each child. The salivary level (low/high) of mutans streptococci was significantly associated with the presence of slightly brown discoloured (C1), clearly brown discoloured (C2) and cavitated (C3) pits and fissures. The odds ratios were 1.5 (P<0.01) for C1, 2.5 (P<0.001) for C2 and 5.0 (P<0.001) for C3 pits and fissures. The findings are consistent with the hypothesis that brown discolouration indicates caries. Furthermore, the findings suggest that this type of discolouration at elementary school age indicates increased caries activity. PMID- 9870542 TI - Perceptions of dental aesthetics in the United States and Nigeria. AB - The standard dental aesthetic index (DAI) is an orthodontic treatment need index based on perceptions of dental aesthetics in the United States. It is a regression equation that links perceptions of the social acceptability of dental aesthetics with the objective intraoral measurements of ten occlusal traits. For the index to be universally acceptable, the perceptions of dental aesthetics in other countries must be similar to those of the United States. This study was designed to determine whether the perceptions of dental aesthetics of Nigerian students are similar to those of different groups of the US population. The same 25 stimuli (photographs of dental configurations, a subset of the 200 stimuli used in deriving the standard DAI equation) were rated for dental aesthetics by Nigerian students. Their ratings were compared with ratings of the same 25 stimuli by US parents, students and orthodontists. Spearman rank-order correlations ranged from 0.75 to 0.84. These correlations were highly significant (P<0.0001), and showed that the perceptions of dental aesthetics of Nigerian students were very similar to those of the US groups. The study therefore concluded that standard DAI could be used as a screening tool and without modification in epidemiological surveys among Nigerians. PMID- 9870543 TI - Orthodontic treatment need prior to treatment and 5 years postretention. AB - OBJECTIVES: To assess orthodontic treatment need prior to treatment and 5 years postretention in a large university sample. METHODS: A random sample of 920 patients (400 male and 520 females) from the Department of Orthodontics and Oral Biology, University of Nijmegen, the Netherlands, were included in this study. Dental casts were evaluated using the Index of Orthodontic Treatment Need (IOTN) at the pretreatment (TP) and 5-year postretention stages (T5). The mean age at TP was 12.2+/-3.0 years and at T5 21.6+/-3.1 years. At TP and T5 the aesthetic component (AC) and the dental health component (DHC) were assessed. The difference between TP and T5 was compared for males and females and tested by the t-test. The changes in AC, DHC and treatment need categories were described at TP and at T5. RESULTS: On the basis of combined AC and DHC treatment need categories, 83% of the patients fell into the "Definite need" category prior to treatment and 10% of the treated patients were categorised as having "Definite need" 5 years postretention. No significant differences were found between males and females for the change in AC and DHC between TP and T5. CONCLUSIONS: The results indicate that the policy used in the department for patient selection gives priority to patients with a high treatment need. The results also provide a general impression of treatment outcome utilising the IOTN to analyse the change in the treatment need categories. PMID- 9870544 TI - A big gene linked to small eyes encodes multiple Mitf isoforms: many promoters make light work. AB - Among more than 80 different loci related to mouse coat color, microphthalmia associated transcription factor (Mitf) encoded at the mouse microphthalmia locus is one of the most exciting molecules that regulates the development and survival of many cell types, including melanocyte, retinal pigment epithelium (RPE), and mast cells. Mitf and its human homolog MITF consist of at least three isoforms, referred to as Mitf-A/MITF-A, the heart-type Mitf-H/MITF-H, and the melanocyte lineage-specific Mitf-M/MITF-M, respectively. These isoforms differ in the amino terminal domains but share a transactivation domain and a basic helix-loop-helix and leucine-zipper structure that is required for DNA binding and dimerization. MITF-M is exclusively expressed in melanocytes and melanoma cells, but not in other cell types, including RPE cells. In contrast, MITF-A mRNA is widely expressed in many cell types. These three isoform mRNAs are possibly generated by differential usage of the gene promoters and by alternative splicing. We predict that the entire MITF gene spans about 200 kb of DNA. Like MITF-M, MITF-A is able to activate the two melanogenesis gene promoters, tyrosinase and tyrosinase related protein 1. These results suggest that melanogenesis may be regulated by different MITF isoforms in melanocyte and RPE. Possible implications of the multiplicity in Mitf/MITF isoforms are discussed. PMID- 9870545 TI - Stereological studies of the effects of alpha-MSH and cAMP on melanosomes in melanoma cells. AB - The effects of alpha-MSH and cAMP on melanosomes in Cloudman S91 melanoma cells were investigated by modern stereological techniques. Cells were cultured for 4 days in medium containing alpha-MSH or cAMP, harvested at 24 hour intervals; some were frozen for melanin assay and the reminder embedded in Epon for light and electron microscopy. Cellular and melanosomal parameters were estimated by new stereological probes. We found that both stimulators induced increases in nuclear volume, cell volume, and the volume fractions and volumes of premelanosomes (V(Vpm,cell), Vpm) and mature melanosomes (V(Vmm,cell), Vmm), and the number of mature melanosomes (Nmm). Both stimulators also caused declines in the volume of individual mature melanosomes (V(Vimm)), the melanin content per mature melanosome unit volume and the melanin content per individual mature melanosome. The increases in the volume of individual premelanosomes and the number of premelanosomes were only induced by cAMP. The effect cAMP on some parameters occurred 24 hours prior to alpha-MSH and was more marked. The response of premelanosomes to the stimulators was more sensitive than mature melanosomes. These results suggest that both stimulators enhanced melanogenesis by increasing the V(Vpm,cell), V(Vmm,cell), Vpm, Vmm and Nmm. The melanogenic level did not depend on the V(Vimm) and melanin concentration in melanosomes. The maturation of premelanosomes was involved in melanogenesis induced by both stimulators, but, de novo synthesis and enlargement of premelanosomes were only stimulated by cAMP. It imply that exogenous cAMP may affect melanosomes, and hence melanogenesis in quantitatively or qualitatively different ways to alpha-MSH. PMID- 9870546 TI - The erythrophore in the larval and adult dorsal skin of the brown frog, Rana ornativentris: its differentiation, migration, and pigmentary organelle formation. AB - To determine whether or not the erythrophore originates from xanthophores in the dorsal skin of the brown frog, Rana ornativentris, we morphologically examined the differentiation and migration of the two chromatophore types and their pigmentary organelle formation. At an early tadpole stage, three kinds of chromatophores, xanthophores, iridophores, and melanophores, appeared in the subdermis, whereas the erythrophore did so just before the foreleg protrusion stage. By the middle of metamorphosis, most chromatophores other than erythrophores had migrated to the subepidermal space. Erythrophores, which appeared late in the subdermis, proliferated actively there during metamorphosis and finished moving into the subepidermal space by the completion of metamorphosis. Carotenoid vesicles and pterinosomes within the erythrophores and xanthophores showed several significant differences in structure. In xanthophores, carotenoid vesicles were abundant throughout life, whereas those in erythrophores decreased in number with the growth of the frogs. The fibrous materials contained in the pterinosomes were initially scattered but soon formed a concentric lamellar structure. In erythrophores, the lamellar structure began to form at the periphery of the organelles but at the center in xanthophores. In addition, the pterinosomes of erythrophores were uniform in size throughout development, while those of xanthophores showed a tendency to become smaller after metamorphosis. The pterinosomes of xanthophores were significantly larger than those of erythrophores. These findings suggest that an erythrophore is not a transformed xanthophore, although they resemble each other closely in many respects. PMID- 9870547 TI - The inhibitory effect of glabridin from licorice extracts on melanogenesis and inflammation. AB - Glabridin is the main ingredient in hydrophobic fraction of licorice extract affecting on skins. In this study, we investigated inhibitory effects of glabridin on melanogenesis and inflammation using cultured B16 murine melanoma cells and guinea pig skins. The results indicated that glabridin inhibits tyrosinase activity of these cells at concentrations of 0.1 to 1.0 microg/ml and had no detectable effect on their DNA synthesis. Combined analysis of SDS polyacrylamide gel electrophoresis and DOPA staining on the large granule fraction of these cells disclosed that glabridin decreased specifically the activities of T1 and T3 tyrosinase isozymes. It was also shown that UVB-induced pigmentation and erythema in the skins of guinea pigs were inhibited by topical applications of 0.5% glabridin. Anti-inflammatory effects of glabridin in vitro were also shown by its inhibition of superoxide anion productions and cyclooxygenase activities. These data indicated that glabridin is a unique compound possessing more than one function; not only the inhibition of melanogenesis but also the inhibition of inflammation in the skins. By replacing each of hydroxyl groups of glabridin with others, it was revealed that the inhibitory effect of 2'-O-ethyl glabridin was significantly stronger than that of 4'-O-ethyl-glabridin on melanin synthesis in cultured B16 cells at the concentration of 1.0 mg/ml. With replacement of both of two hydroxyl groups, the inhibitory effect was totally lost. Based on these data, we concluded that two hydroxyl groups of glabridin are important for the inhibition of melanin synthesis and that the hydroxyl group at the 4' position of this compound is more closely related to melanin synthesis. PMID- 9870548 TI - Light-induced pigment aggregation in xanthophores of the medaka, Oryzias latipes. AB - The response mechanism of medaka xanthophores to light was examined at the cellular level. Innervated and denervated xanthophores of adult medakas responded to light (9,000 lux) within 30 sec by pigment aggregation, and this aggregation was not mediated through alpha-adrenoceptors on the cell membrane. Maximum sensitivity to light was at wavelengths of 410-420 nm, and the direct effect of light was reversible. Xanthophore responsiveness to light in summer was higher than that in winter. Ca2+ and calmodulin were not involved in the response, but rather, an important role for cAMP and phosphodiesterase (PDE) was suggested. It seems likely that photoreception by visual pigment which is sensitive to light at wavelengths of 410-420 nm increases PDE activity, probably via a G-protein, such as occurs with visual cells in the retina, which causes a decrease in levels of cytosolic cAMP, in turn leading to pigment aggregation within medaka xanthophores. PMID- 9870549 TI - Involvement of pteridines in the body coloration of the isopod Armadillidium vulgare. AB - The pteridine content was measured as a function of age in Armadillidium vulgare, and the fine structure of the pteridine-containing granules in the integument was examined in relation to pteridine content. Yellow chromatophores are an essential component of the cream-markings, which are a defining feature of the female A. vulgare. Four kinds of pteridines in the integument including a yellow pigment (sepiapterin) were determined by HPLC. The body color of the red phenotype of A. vulgare varies from dark red to yellowish red and was formerly thought to be due to the quality and quantity of ommochrome pigment. Our analysis of the pteridine content in the integument of this phenotype revealed a significant change in sepiapterin content per body weight with age. Sepiapterin content per body weight decreased gradually with age, while that of biopterin tended to increase with age. Ultrastructural observations of the pigment granules in the yellow chromatophores revealed a corresponding change in the fine structure of pigment granules. In the older adults, some of the electron-dense fibrous materials in the pteridine-containing granules was concentrically arranged, and in the younger adults, most of pteridine-containing granules were electron-lucent. The role of pteridine quality in determining the structure of pteridine-containing granules is discussed. PMID- 9870550 TI - Transient expression of high molecular weight, heat sensitive, trypsin-resistant form of tyrosinase in B-16 melanoma cells. AB - High molecular weight forms of tyrosinase have been found to be expressed during spontaneous remelanization of the amelanotic B-16 melanoma cells in culture as well as in melanotic tumors formed from amelanotic melanoma cells grown in C57BL/6J mice. Overnight extraction of the crude melanosomal fractions from such tumors and cultured melanoma cells reveal the presence of an additional DOPA-MBTH positive band well below the stacking gel. This band has been found to be alpha PEP7 (antibody specific for tyrosinase) positive and alpha-PEP1 (antibody specific for TRP-1) negative on Western blot analysis. Heat treatment at 60 degrees C for 60 min results in the loss of this band and considerable loss of activity of the melanosomal extract. Trypsin treatment of these melanosomal extracts resulted in a minor change in the mobility of the high molecular weight band. SDS-PAGE under reduced conditions followed by Western blotting revealed that the high molecular weight band was lost and not detected by alpha-PEP7 or alpha-PEP1. These findings indicate that high molecular weight, heat sensitive and trypsin resistant forms of tyrosinase are transiently expressed in B-16 melanoma cells and tumors that are initiating remelanization following phenotypic drift towards the amelanotic state. PMID- 9870551 TI - Serum malondialdehyde: possible use for the clinical management of chronic hepatitis C patients. AB - Serum lipid peroxidation products are increased in inflammatory liver disease and, as we previously reported, also in chronic hepatitis C. We have performed a specific assay of malondialdehyde, the reported most abundant product of lipid peroxidation, in serum of twenty four chronic hepatitis C patients, before, during, and after interferon treatment. Liver biopsies were performed in each patient before and after interferon treatment. The results show higher serum malondialdehyde values in chronic hepatitis C patients than healthy subjects (n = 68) before interferon treatment (p < .001). Mean value of serum malondialdehyde levels after interferon treatment was significantly lower than before it (p < .002). Associating the histopathological findings in each of the 48 biopsies performed, with serum malondialdehyde and alanine aminotransferase activity levels, of the sample obtained the same day of biopsy, a much better correspondence with the histopathological severity was observed for malondialdehyde concentration than for alanine aminotransferase activity. These levels decreased significantly after interferon treatment. However, when the patients were grouped in responding (group I; n = 9) and non-responding (group II; n = 15) to interferon treatment, according to the histopathological findings before and after interferon, the values of group I before interferon treatment were significantly higher than group II (p < .03). Thus, a potential predictive value could be ascribed to the serum malondialdehyde levels before interferon treatment in these patients. We propose the utility of the specific assay of malondialdehyde for the clinical management of chronic hepatitis C patients. PMID- 9870552 TI - Redox regulation of wound healing? NF-kappaB activation in cultured human keratinocytes upon wounding and the effect of low energy HeNe irradiation. AB - The complex process of wound healing as well as the signaling systems orchestrating this intricate process remain incompletely defined. Using human keratinocytes in primary culture, we sought to characterize their NF-kappaB responses to wounding alone or in combination with other treatments. We initially characterized these cultured human keratinocytes responses to known NF-kappaB activators (PMA, TNF-alpha and IL-1) using two different assays, immunohistochemistry and electrophoretic mobility shift (EMSA). After eliciting the expected NF-kappaB responses, we applied these same assays to assess responses to either wounding or HeNe irradiation alone. The results obtained indicated that only a modest/sporadic activation of NF-kappaB was elicited by these which was only detectable using immunohistochemistry. When the combination of wounding and HeNe irradiation on NF-kappaB status was assessed, a marked, localized activation of NF-kappaB in keratinocytes along the wound edge was found. Treatment induced NF-kappaB activation (e.g., wounding, HeNe irradiation and combined wounding and HeNe irradiation) was abrogated by pyrrolidine dithiocarbamate (PDTC) which inhibits NF-kappaB activation through an as yet incompletely understood (antioxidant?) mechanism. These data therefore suggest that NF-kappaB and oxidation mediated changes in its activation state likely play important roles in normal cutaneous wound healing. PMID- 9870553 TI - Modulation of UV-light-induced skin inflammation by D-alpha-tocopherol and L ascorbic acid: a clinical study using solar simulated radiation. AB - OBJECTIVE: In this clinical trial we studied whether oral supplementation with D alpha-tocopherol (alpha-Toc), L-ascorbic acid (Asc), or alpha-Toc combined with Asc influenced the solar simulated radiation (SSR) induced skin inflammation in healthy volunteers. METHODS: We investigated the following groups in a prospective, randomized and placebo controlled study: Group (1) alpha-Toc 2 g/day, group (2) Asc 3 g/day, group (3) alpha-Toc 2 g/day combined with Asc 3 g/day, and group (4) placebo. Before and 50 days after supplementation we analyzed alpha-Toc and Asc concentrations in keratinocytes. The dose response curve of UV erythema was determined by reflectance spectrophotometry and the minimal erythema dose (MED) by visual grading before and after supplementation. RESULTS: 50 days after supplementation alpha-Toc keratinocyte levels were increased in groups (1) and (3), Asc concentrations were elevated in groups (2) and (3), and the a/gamma-Toc ratio increased in groups (1) and (3). The dose response curve of UVR induced erythema showed a significant flattening and the MED increased from 103 +/- 29 mJ/cm2 (before supplementation) to 183 +/- 35 mJ/cm2 (after supplementation) in group (3), while there were no significant changes in groups (1) and (2) after vitamin supplementation. CONCLUSION: Alpha Toc and Asc act synergistically in suppression of the sunburn reaction. PMID- 9870554 TI - Interaction between sodium ascorbate and dopamine. AB - The interaction between sodium ascorbate and dopamine was investigated by three different parameters: radical intensity, prooxidant action, and cytotoxicity induction. Sodium ascorbate and dopamine produced the doublet and quartet ESR signals under alkaline conditions (pH 8.0-9.5), respectively. Addition of increasing concentrations of sodium ascorbate completely scavenged the dopamine radical and replaced the latter with its own radical. Similarly, dopamine slightly, but significantly reduced the radical intensity of sodium ascorbate. These two compounds stimulated the methionine oxidation and hydrogen peroxide generation in culture medium, but in combination, their stimulation activities were weakened. Both of these two compounds dose-dependently reduced the viable cell number of human oral squamous carcinoma HSC-4 cells, and their cytotoxic activity was significantly reduced by catalase. When these two compounds were mixed together before adding to HSC-4 cells, both of their cytotoxic activities were diminished. The present study demonstrates the interaction between sodium ascorbate and dopamine, which might modify their biological activities and generation of nerve disorders such as Parkinson's disease. PMID- 9870555 TI - Reactive oxygen species activate focal adhesion kinase, paxillin and p130cas tyrosine phosphorylation in endothelial cells. AB - Reactive oxygen species (ROS), particularly hydroxyl radical (HO*), increase neutrophil adherence to hypoxanthine-xanthine oxidase (HX-XO)-treated human umbilical vein endothelial cells (HUVEC) in culture. This adherence is inhibited by the tyrosine kinase inhibitors genistein (30 microM) and herbimycin A (0.9 microM), suggesting the involvement of tyrosine kinase. Phosphorylation of several HUVEC proteins in the range of 120-130 and 70 kDa was found to depend on the XO concentration and stimulation time. This phosphorylation was inhibited by the antioxidants dimethylthiourea (DMTU, 0.75 to 7.5 mM) and pentoxifylline (Ptx, 0.1 mM), and by the iron chelators desferrioxamine (DF, 1 mM) and hydroxybenzyl ethylene diamine (HBED, 0.5 mM), suggesting the involvement of HO*. Three tyrosine-phosphorylated proteins, focal adhesion kinase (p125FAK), paxillin (PAX) and p130cas were isolated and characterized by immunoprecipitation and western blotting. Antioxidants and iron chelators reduced their phosphorylation. HUVEC treated with ROS for 15 min showed actin stress fiber formation. Cytochalasin D (5 microM) inhibited tyrosine phosphorylation and PMN-HUVEC adherence, showing the importance of cytoskeleton integrity in these two functions. In conclusion, HO*, which is involved in increased PMN-HUVEC adhesion, also increases tyrosine phosphorylation on three major cytoskeleton proteins which seem to play a role in this adhesion. PMID- 9870556 TI - Effects of oxygen on 3-hydroxyanthranilate oxidase of the kynurenine pathway. AB - Iron containing 3-Hydroxyanthranilate oxidase (3HAO) converts 3 hydroxyanthranilate (3HAA) and dioxygen into a precursor which spontaneously converts to quinolinic acid (QA). 3HAO participates in de novo biosynthesis of NAD in mammalian kidney and liver, and it is present in low concentrations in brain where its function is controversial. However, QA increases in spinal fluid and is associated with convulsions in AIDS dementia, Huntington's disease, and CNS inflammation. QA is a known N-methyl, D-aspartate receptor agonist and excitotoxin that causes convulsions when injected into the brain. Hyperbaric oxygen (HBO) also causes convulsions and we investigated the interrelationships among the stimulating and toxic effects of oxygen and the role of iron in vitro using rat liver enzyme which is reported to be identical to brain enzyme and is more abundant. 3HAO requires dioxygen as a substrate but it was inactivated approximately 40% by 5.2 atm HBO in vitro in 15 min. The apparent Km was 2.6 x 10(-4) M for oxygen and 5 x 10(-5) M for 3HAA, and these values did not change for enzyme that was half-inactivated by HBO oxygen. Thus, oxygen-inactivation appears to be all-or-none for individual enzyme molecules. Freshly prepared enzyme was activated about 3-fold by incubation with acidic iron. Iron-staining of 3HAO, separated by gel electrophoresis after partial purification by FPLC, showed that loss of iron and loss of enzyme activity during HBO exposure were correlated. The apparent oxygen Km of 3HAO is far higher than the oxygen concentration in brain cells. Thus, 3HAO is capable of being stimulated initially in animals breathing HBO, and subsequently of being inactivated with potential significance for brain QA and convulsions. PMID- 9870557 TI - Biomarkers of oxidative stress are significantly elevated in Down syndrome. AB - There is convincing epidemiological and in vitro evidence of chronic oxidative stress in individuals with Down syndrome (DS). These individuals develop Alzheimer like changes in the brain in their 30s and 40s. The incidence of autoimmune diseases and cataracts is significantly increased, and the overall ageing process is accelerated. In vitro studies show that impaired viability of DS neurons may be amended by simple chemical antioxidants, such as vitamin E, BHT and propyl gallate, clearly indicative of oxyl radical involvement. However, because of the lack of in vivo experiments, the role of oxidative stress in DS remains controversial. We report here on the results of the chemical analyses of urine samples of 166 individuals, where DS subjects were matched by their siblings. The levels of 8-hydroxy-2'-deoxyguanosine (2.35 +/- 1.69 in DS vs. 1.35 +/- 1.04 in controls, P = 0.00011), a biomarker of oxidative damage to DNA, and malondialdehyde (0.255 +/- 0.158 in DS vs. 0.204 +/- 0.128 in controls, P = 0.033), a biomarker of lipid peroxidation, are significantly elevated in individuals with DS. Dietary influences failed to show any significant correlation with the oxidative stress biomarkers. These results provide direct evidence for increased oxidative stress in individuals with DS. PMID- 9870558 TI - Covalent modifications of aminophospholipids by 4-hydroxynonenal. AB - Lipid oxidation is implicated in a wide range of pathophysiological disorders, which leads to reactive compounds such as aldehydes. Among them 4-hydroxynonenal (4-HNE) reacts strongly with the NH2 groups of amino acids and forms mainly Michael adducts and minor Schiff-base adducts. Such reactions occur also with compounds containing thiol groups. No data are available describing 4-HNE interactions with amino-phospholipids. To investigate such a possibility, 4-HNE was incubated with either phosphatidylethanolamine (PE) or phosphatidylserine (PS) in an aqueous-organic biphasic system and the resulting products were identified by liquid chromatography-mass spectrometry (LC-MS). Our study points out the potential capacity of 4-HNE to react with phospholipids containing amino groups and particularly PE. The main resulting compounds found were a Michael adduct plus a minor Schiff base adduct, which was partly cyclized as a pyrrole derivative via a loss of water. Its stabilization as a pyrrole derivative allows to differentiate 4-HNE from the other aldehydes generated via lipid oxidation (e.g., malondialdehyde, 2-nonenal) that lack the 4-hydroxyl group. Their formation seems not to be affected when the pH varies from 6.5 to 8.5. Surprisingly, PS reacted poorly producing only a small amount of Michael adduct, the Schiff-base adduct being nondetectable. We conclude that such adducts, if they are formed in cell membranes, could alter the phospholipase-dependent cell signaling. PMID- 9870559 TI - Antioxidant and pro-oxidant actions of flavonoids: effects on DNA damage induced by nitric oxide, peroxynitrite and nitroxyl anion. AB - Antioxidant and pro-oxidant activities of flavonoids have been reported. We have studied the effects of 18 flavonoids and related phenolic compounds on DNA damage induced by nitric oxide (NO), peroxynitrite, and nitroxyl anion (NO-). Similarly to our previous findings with catecholamines and catechol-estrogens, DNA single strand breakage was induced synergistically when pBR322 plasmid was incubated in the presence of an NO-releasing compound (diethylamine NONOate) and a flavonoid having an ortho-trihydroxyl group in either the B ring (e.g., epigallocatechin gallate) or the A ring (e.g., quercetagetin). Either NO or any of the above flavonoids alone did not induce strand breakage significantly. However, most of the tested flavonoids inhibited the peroxynitrite-mediated formation of 8 nitroguanine in calf-thymus DNA, measured by a new HPLC-electrochemical detection method, as well as the peroxynitrite-induced strand breakage. NO- generated from Angeli's salt caused DNA strand breakage, which was also inhibited by flavonoids but at only high concentrations. On the basis of these findings, we propose that NO- and/or peroxynitrite could be responsible for DNA strand breakage induced by NO and a flavonoid having an ortho-trihydroxyl group. Our results indicate that flavonoids have antioxidant properties, but some act as pro-oxidants in the presence of NO. PMID- 9870560 TI - Generation of superoxide anion by mitochondria and impairment of their functions during anoxia and reoxygenation in vitro. AB - A small portion of the oxygen consumed by aerobic cells is converted to superoxide anion at the level of the mitochondrial respiratory chain. If produced in excess, this harmful radical is considered to impair cellular structures and functions. Damage at the level of mitochondria have been reported after ischemia and reperfusion of organs. However, the complexity of the in vivo system prevents from understanding and describing precise mechanisms and locations of mitochondrial impairment. An in vitro model of isolated-mitochondria anoxia reoxygenation is used to investigate superoxide anion generation together with specific damage at the level of mitochondrial oxidative phosphorylation. Superoxide anion is detected by electron paramagnetic resonance spin trapping with POBN-ethanol. Mitochondrial respiratory parameters are calculated from oxygen consumption traces recorded with a Clark electrode. Respiring mitochondria produce superoxide anion in unstressed conditions, however, the production is raised during postanoxic reoxygenation. Several respiratory parameters are impaired after reoxygenation, as shown by decreases of phosphorylating and uncoupled respiration rates and of ADP/O ratio and by increase of resting respiration. Partial protection of mitochondrial function by POBN suggests that functional damage is related and secondary to superoxide anion production by the mitochondria in vitro. PMID- 9870562 TI - Ketosis (acetoacetate) can generate oxygen radicals and cause increased lipid peroxidation and growth inhibition in human endothelial cells. AB - Elevated level of cellular lipid peroxidation can increase the incidence of vascular disease. The mechanism by which ketosis causes accelerated cellular damage and vascular disease in diabetes is not known. This study was undertaken to test the hypothesis that elevated levels of ketone bodies increase lipid peroxidation in endothelial cells. Human umbilical venous endothelial cells (HUVEC) were cultured for 24 h at 37 degrees C with ketone bodies (acetoacetate, beta-hydroxybutyrate). Acetoacetate, but not beta-hydroxybutyrate, caused an increase in lipid peroxidation and growth inhibition in cultured HUVEC. To determine whether ketone bodies generate oxygen radicals, studies using cell-free buffered solution were performed. They showed a significant superoxide dismutase (SOD) inhibitable reduction of cytochrome C by acetoacetate, but not by beta hydroxybutyrate, suggesting the generation of superoxide anion radicals by acetoacetate. Additional studies show that Fe2+ potentiates oxygen radical generation by acetoacetate. Thus, elevated levels of ketone body acetoacetate can generate oxygen radicals and cause lipid peroxidation in endothelial cells, providing a possible mechanism for the increased incidence of vascular disease in diabetes. PMID- 9870561 TI - Crucial role of apopain in the peroxynitrite-induced apoptotic DNA fragmentation. AB - Peroxynitrite, a cytotoxic oxidant formed in the reaction of superoxide and nitric oxide is known to cause programmed cell death. However, the mechanisms of peroxynitrite-induced apoptosis are poorly defined. The present study was designed to characterize the molecular mechanisms by which peroxynitrite induces apoptosis in HL-60 cells, with special emphasis on the role of caspases. Peroxynitrite induced the activation of apopain/caspase-3, but not ICE/caspase-1 as measured by the cleavage of fluorogenic peptides. Considering the short half life of peroxynitrite and the kinetics of caspase-3 activation (starting 3-4 h after peroxynitrite treatment), the enzyme is not likely to become activated directly by the oxidant. Caspase-3 activation proved to be essential for DNA fragmentation, because pretreatment of the cells with the specific tetrapeptide inhibitor DEVD-fmk completely blocked peroxynitrite-induced DNA fragmentation. Peroxynitrite-induced cytotoxicity was also significantly altered by the inhibition of caspase-3, whereas phosphatidylserine exposure was unaffected by DEVD-fmk treatment. Because many of the effects of peroxynitrite are mediated by poly(ADP-ribose) synthetase (PARS) activation, we have also investigated the effect of PARS-inhibition on peroxynitrite-induced apoptosis. We have found that PARS-inhibition modulates peroxynitrite-induced apoptotic DNA fragmentation in the HL-60 cells. The effect of the PARS inhibitors, 3-aminobenzamide and 5-iodo-6 amino-1,2-benzopyrone were dependent on the concentration of peroxynitrite used. While PARS-inhibition resulted in increased DNA-fragmentation at low doses (15 microM) of peroxynitrite, a decreased DNA-fragmentation was found at high doses (60 microM) of peroxynitrite. PARS inhibition negatively affected viability as determined by flow cytometry. These data demonstrate the crucial role of caspase 3 in mediating apoptotic DNA fragmentation in HL-60 cells exposed to peroxynitrite. PMID- 9870563 TI - Caloric restriction prevents age-associated accrual of oxidative damage to mouse skeletal muscle mitochondria. AB - The purpose of this study was to understand the nature of the causes underlying the senescence-related decline in skeletal muscle mass and performance. Protein and lipid oxidative damage to upper hindlimb skeletal muscle mitochondria was compared between mice fed ad libitum and those restricted to 40% fewer calories- a regimen that increases life span by approximately 30-40% and attenuates the senescence-associated decrement in skeletal muscle mass and function. Oxidative damage to mitochondrial proteins, measured as amounts of protein carbonyls and loss of protein sulfhydryl content, and to mitochondrial lipids, determined as concentration of thiobarbituric acid reactive substances, significantly increased with age in the ad libitum-fed (AL) C57BL/6 mice. The rate of superoxide anion radical generation by submitochondrial particles increased whereas the activities of antioxidative enzymes superoxide dismutase, catalase, and glutathione peroxidase in muscle homogenates remained unaltered with age in the AL group. In calorically-restricted (CR) mice there was no age-associated increase in mitochondrial protein or lipid oxidative damage, or in superoxide anion radical generation. Crossover studies, involving the transfer of 18- to 22-month-old mice fed on the AL regimen to the CR regimen, and vice versa, indicated that the mitochondrial oxidative damage could not be reversed by CR or induced by AL feeding within a time frame of 6 weeks. Results of this study indicate that mitochondria in skeletal muscles accumulate significant amounts of oxidative damage during aging. Although such damage is largely irreversible, it can be prevented by restriction of caloric intake. PMID- 9870564 TI - Effect of caffeic acid on tert-butyl hydroperoxide-induced oxidative stress in U937. AB - Nonvitamin phenolic compounds are ubiquitous in food plants and therefore potentially present in human plasma in a diet-dependent concentration. The aim of this study was to evaluate the ability of caffeic acid, a phenolic acid with antioxidant activity, to affect cellular response in U937 human monocytic cells to t-butyl hydroperoxide-induced oxidative stress. In our experimental conditions caffeic acid was incorporated into cells without any cytotoxic effect. Caffeic acid-treated cells showed an increased resistance to oxidative challenge, as revealed by an higher percent of survival and the maintenance of an higher proliferative capacity in respect to control cells. This effect seems to be due to the ability of caffeic acid to reduce glutathione depletion and to inhibit lipid peroxidation during tBOOH treatment. It can be concluded that caffeic acid exerts an antioxidant action inside the cell, responsible for the observed modulation of the cellular response to oxidative challenge. Due to its presence in the diet, therefore, caffeic acid may play a role in the modulation of oxidative processes in vivo. PMID- 9870565 TI - Oxidative stress causes a general, calcium-dependent degradation of mitochondrial polynucleotides. AB - Oxidative stress has many effects on biological cells, including the modulation of gene expression. Reactive oxygen species are known to up-regulate and down regulate RNA expression in prokaryotic and eukaryotic cells. We have previously reported that a preferential and calcium-dependent down-regulation of mitochondrial RNAs occurs when HA-1 hamster fibroblasts are exposed to hydrogen peroxide. Here we extend these studies to determine whether this down-regulation is specific to mitochondria RNA or involves general polynucleotide degradation. Degradation and associated decreases in the levels of 16S mitochondrial rRNA following exposure of cells to 400 microM hydrogen peroxide were found to be dependent on calcium at 2 and 5 h. Degradation of mitochondrial genomic DNA was also observed following peroxide exposure, and occurred at similar time points as for mitochondrial RNA degradation. As with mitochondrial RNA degradation, this mitochondrial genomic DNA degradation was dependent on calcium. These results indicate that there is a general, calcium-dependent degradation of mitochondrial polynucleotides following exposure of HA-1 fibroblasts to oxidative stress, and suggest that a dramatic shut-down in mitochondrial biosynthesis is an early-stage response to oxidative stress. PMID- 9870566 TI - Laparoscopic gastric bypass. PMID- 9870567 TI - Combined low-pressure laparoscopic gastric banding. PMID- 9870568 TI - Biliointestinal bypass: another surgical option. PMID- 9870569 TI - The role of bisphosphonates in the treatment of painful metastatic bone disease: a review of phase III trials. AB - Metastatic bone disease is a frequent cause of morbidity in advanced cancer patients with a subsequent high incidence of skeletal complications (fractures, hypercalcemia, spinal cord compression) and severe pain. The osteolytic process is mainly characterized by an osteoclastic activity of bone resorption and inflammatory activity provoked by various cytokines and prostaglandins. Bisphosphonates represent a new class of drugs with inhibitory activity on bone resorption and on inflammatory processes which revealed themselves to be efficacious in a series of clinical conditions such as tumour-induced hypercalcemia, Paget's disease, osteoporosis and metastatic bone disease. The aim of this review of the literature is to show the analgesic efficacy of the different bisphosphonates in phase III studies carried out on patients with metastatic bone disease. Medline and Cancerlit database from January 1984 to February 1998 have been considered. From the analysis of the published studies it appears that bisphosphonates and, in particular, intravenous Disodium Pamidronate, are not only able to slow down the progression of the disease and to reduce the onset of skeletal complications but also have an analgesic effect and the possibility of improving the quality of life, above all in patients with osteolytic metastases due to breast cancer and multiple myeloma. Bisphosphonates represent a further valid therapy to add to an already consolidated list of therapies such as radio, chemo and endocrine therapy, analgesic drugs, orthopaedic and physiatric in the pain management of patients with bone metastases. These drugs meet with the patients' compliance, are well-tolerated as well as having a good cost/efficacy profile. It still remains to be seen if the newer and more potent bisphosphonates such as Ibandronate and Zoledronate can be administered differently from the intravenous route such as by mouth or by patch which are readily accepted by the patient and, moreover, if these more potent drugs are able to prevent or delay the onset and/or the progression of bone metastases. PMID- 9870570 TI - Effect of peripheral nerve injury on cGMP and nitric oxide synthase levels in rat dorsal root ganglia: time course and coexistence. AB - Using the indirect immunofluorescence method, the distribution of cyclic GMP (cGMP) and nitric oxide synthase (NOS) was investigated in lumbar 5 dorsal root ganglia (DRGs) of untreated rats 1, 3 and 7 days following sciatic nerve section (axotomy). Untreated and axotomized (7 days) rats were also studied after perfusion with the NO donor sodium nitroprusside (SNP). Moreover, rats were injected with carrageenan lambda into the unilateral hindpaw and studied after 6 h, 1 day or 2 days. An increase in the number of cGMP-positive satellite cell profiles was found in axotomized DRGs at 3 days with lower numbers after 7 days. In contrast, no change in cGMP-like immunoreactivity (LI) in satellite cell profiles was detected 1 day after axotomy or 6h, 1 day or 2 days after inflammation, as compared to controls. Axotomy induced a marked increase in the percentage of NOS-immunoreactive (IR) neuron profiles in the ipsilateral DRGs as follows: 3.0% at 1 day, 15% at 3 days and 25% at 7 days, whereas no significant change was found in the expression of NOS-LI in the inflamed DRGs as compared to untreated DRGs. Between 15 and 20% of all NOS-positive neuron profiles were surrounded by, or in partial contact with, cGMP-IR satellite cells in controls 1 and 3 days after axotomy, whereas the corresponding figure was around 5% after 7 days. After SNP perfusion 60-70% of all DRG neuron profiles were partly or totally associated with cGMP-positive satellite cell profiles, with no significant difference between untreated and axotomized ganglia. The nerve injury induced, parallel upregulation of NOS in DRG neurons and cGMP in satellite cells in the initial phase after axotomy suggests an involvement of NO as a signalling molecule between neurons and satellite cells in DRGs, especially after peripheral nerve injury, perhaps exerting a survival effect as recently proposed by Thippeswamy and Morris (1997). PMID- 9870571 TI - Lipopolysaccharide-induced hyperalgesia of intracranial capsaicin sensitive afferents in conscious rats. AB - Migraineous and non-migraineous headache is reported to be at highest intensity after an infection. This study investigated whether activation of the immune system can induce hyperalgesia in intracranial capsaicin sensitive afferents. The effects of intraperitoneal injected lipopolysaccharides (LPS) on behavior and c fos expression in the trigeminal nucleus caudalis layer I, II (TNC I,II) elicited by intracisternally applied capsaicin were studied. Low concentrations of LPS potentiated capsaicin-induced immobilization behavior without affecting c-fos expression in the TNC I,II. Large amounts of LPS however increased the number of capsaicin-induced c-fos positive cells in the TNC I,II. These effects of LPS on capsaicin sensitive afferents are probably mediated by cytokines that act at peripheral vagal nerves, central brain regions or via direct actions of cytokines on capsaicin sensitive afferent nerve terminals. The hyperalgesic action of LPS on intracranial trigeminal and possibly other capsaicin sensitive afferents of the head may explain why different types of infections are accompanied by headache and why migraineous and non-migraineous headache is of highest intensity after an infection. PMID- 9870572 TI - Pathological mechanisms implicated in localized female trapezius myalgia. AB - Myalgia localized to the neck and shoulder in women is a growing problem both in the general population and in the industrial world. The aim of this study was to investigate the mechanisms involved in work-related myalgia. In 21 women (age, 38.7+/-5.5 years), muscle biopsies were obtained from the upper part of the trapezius and the morphologic and metabolic characteristics of muscle fibres were analyzed. The patients indicated the number of painful areas on a pain drawing and the intensity of pain was assessed using a visual analogue scale (VAS). Two groups were formed on the basis of the median values: lower pain level and higher pain level. Trapezius muscles were characterized by the large size of type I fibres and the low capillary to fibre area ratio for both type I and type IIA fibres. Patients with the highest pain scores had the lowest capillary to fibre area ratio for type I fibres (coefficient correlation r = -0.45 and P < 0.05). Moreover, the proportion of cytochrome c oxidase (COX)-negative fibres seen in the cross-sections was significantly higher in the group of patients which had the higher pain and more painful areas than in the group of patients with lower pain level and painful areas (P < 0.05). The significant increase (P < 0.05) of the size of the type I fibres in trapezius myalgia point to the special strain imposed upon type I muscle fibres during work tasks. Cytochrome oxidase c deficiency which is indicative of an energy crisis within muscle cells and the low capillary to fibre area ratio which might impair oxygen delivery and removal of metabolites in the working muscles are both associated with pain in the trapezius muscle. PMID- 9870574 TI - Size is everything--large amounts of information are needed to overcome random effects in estimating direction and magnitude of treatment effects. AB - Variability in patients' response to interventions in pain and other clinical settings is large. Many explanations such as trial methods, environment or culture have been proposed, but this paper sets out to show that the main cause of the variability may be random chance, and that if trials are small their estimate of magnitude of effect may be incorrect, simply because of the random play of chance. This is highly relevant to the questions of 'How large do trials have to be for statistical accuracy?' and 'How large do trials have to be for their results to be clinically valid?' The true underlying control event rate (CER) and experimental event rate (EER) were determined from single-dose acute pain analgesic trials in over 5000 patients. Trial group size required to obtain statistically significant and clinically relevant (0.95 probability of number needed-to-treat within -/+0.5 of its true value) results were computed using these values. Ten thousand trials using these CER and EER values were simulated using varying group sizes to investigate the variation due to random chance alone. Most common analgesics have EERs in the range 0.4-0.6 and CER of about 0.19. With such efficacy, to have a 90% chance of obtaining a statistically significant result in the correct direction requires group sizes in the range 30 60. For clinical relevance nearly 500 patients are required in each group. Only with an extremely effective drug (EER > 0.8) will we be reasonably sure of obtaining a clinically relevant NNT with commonly used group sizes of around 40 patients per treatment arm. The simulated trials showed substantial variation in CER and EER, with the probability of obtaining the correct values improving as group size increased. We contend that much of the variability in control and experimental event rates is due to random chance alone. Single small trials are unlikely to be correct. If we want to be sure of getting correct (clinically relevant) results in clinical trials we must study more patients. Credible estimates of clinical efficacy are only likely to come from large trials or from pooling multiple trials of conventional (small) size. PMID- 9870573 TI - The involvement of bulbospinal pathways in fentanyl-induced inhibition of spinal withdrawal reflexes in the decerebrated rabbit. AB - The selective opioid OP3(mu)-receptor agonist fentanyl was administered via the intravenous, intrathecal and intraventricular routes to decerebrated rabbits in doses from 1-30 microg/kg. Reflexes evoked in medial gastrocnemius motoneurones by electrical stimulation of the sural nerve were depressed by fentanyl given by all three routes. The opioid was most potent when given intrathecally and least potent when given into the fourth ventricle. Blockade of spinal alpha2 adrenoceptors by intrathecal RX 821002 (100 microg) reduced the effectiveness of intrathecal and low (<3 microg/kg) intravenous doses of fentanyl, but did not affect or enhanced responses to high intraventricular and intravenous doses. Spinalization reduced the effectiveness of intrathecal and intravenous fentanyl and abolished inhibition from intraventricular dosing. These data show that fentanyl acts in the spinal cord and in the brain stem to suppress spinal reflexes, although very high doses were required for effects from the latter site. It appears that low intravenous doses of fentanyl act mainly in the spinal cord and that increasing the dosage recruits descending inhibition. The results of alpha2-adrenoceptor blockade indicate that the spinal inhibitory effects of opioids are enhanced by an interaction with endogenous noradrenaline in the spinal cord. Thus, the full expression of the spinal inhibitory effects of fentanyl is dependent on brain stem cell groups, either as a source of noradrenaline input to the spinal cord, or as a site from which opioids can activate descending inhibitory systems. PMID- 9870575 TI - The control of severe cancer pain by continuous intrathecal infusion and patient controlled intrathecal analgesia with morphine, bupivacaine and clonidine. AB - The management of severe cancer pain may be problematic in spite of recent advances in pain management. A small percentage of patients with severe intractable pain and/or intractable side effects may require more aggressive interventional pain management strategies including the administration of medications continuously by the intrathecal route. A variety of medications, including morphine, bupivacaine, and clonidine, may be used intrathecally for the control of cancer pain. Optimal use of these medications requires individual titration to the patient's needs. We describe a case of severe cancer pain where these medications were used successfully by continuous intrathecal infusion and patient controlled intrathecal analgesia. PMID- 9870576 TI - Comments on Eisenberg and Pud, PAIN, 74 (1998) 337-339. PMID- 9870577 TI - Comments on Pfingsten et al., PAIN, 73 (1997) 77-85. PMID- 9870578 TI - Reply to Wallis et al., PAIN 73 (1997) 15-22. PMID- 9870579 TI - Comments on Gould, PAIN, 74 (1998) 341-343. PMID- 9870580 TI - Gene expression regulated by gene VI of caulimovirus: transactivation of downstream genes of transcripts by gene VI of peanut chlorotic streak virus in transgenic tobacco. AB - Here we document that the gene VI product of peanut chlorotic streak virus (PClSV), a newly characterized member of the group, transactivates the translation of dicistronic transcripts. Dicistronic expression units have been analyzed both in protoplast transient expression experiments and in transgenic tobacco plants. Transgenic plants containing a dicistronic transcription unit (PClSV-gene VII-GUS) under the control of PClSV full-length transcript promoter with its long leader sequence show a relatively high abundance of the expected transcript but very little, or no, GUS activity. However, high GUS activity is found when gene VI protein is then provided by subsequent infection with PClSV. The efficient translation of polycistronic mRNAs mediated by gene VI of caulimovirus has potential value in product engineering of plants. PMID- 9870581 TI - The position of the ZEBRA activation domain does not influence its biological activity. AB - Epstein-Barr virus (EBV) is a human herpesvirus which latently infects B lymphocytes. EBV encodes a unique transcriptional activator, known as ZEBRA, which can disrupt viral latency in B cells and induce lytic viral replication. Furthermore, ZEBRA has been shown to bind at the EBV origin of lytic replication, and is necessary for viral DNA replication to occur. Previously we demonstrated that heterologous activation domains can fully substitute for the ZEBRA activation domain. Here we extend those results by showing that the position of the ZEBRA activation domain or a heterologous replacement domain does not influence its ability to function in the disruption of EBV latency. In this study three novel clones were constructed in which the ZEBRA activation region was repositioned to the carboxy terminus of the protein. These mutants were used to demonstrate that the ability of ZEBRA's wild type domain to function in the complex biological process of virus activation is not compromised by altering its position within the protein. PMID- 9870582 TI - The muskrat (Ondatra zibethicus) as a new reservoir for puumala-like hantavirus strains in Europe. AB - We have used an indirect immunofluorescent assay (IFA) and reverse transcription PCR (RT-PCR) to screen the sera and tissues of muskrats (Ondatra zibethica) caught in the northwest of Brandenburg and in the northeast of Saxony-Anhalt, Germany, for hantavirus infection. Kidney and/or lung tissue from 6 (3.1%, CI = 1.1-6.5%) out of 197 muskrats were found to be positive for genomic sequences of hantavirus by RT-PCR. We could also demonstrate that 14 (5%, CI = 2.9-8.7%) out of 266 muskrat's sera available for testing contained hantavirus-specific antibodies in IFA. Thus, a total of 8% of the investigated muskrat population was found to be positive for hantavirus infection by RT-PCR and IFA. None of the animals was found positive in both tests. Further analysis of the RT-PCR amplified fragments by genomic sequencing revealed sequences mostly related to the puumala (PUU) S segment sequence of the Hallnas B1 hantavirus strain (97-99% similarity). Our data therefore demonstrate that Ondatra zibethicus serves as an additional reservoir for puumala-like hantavirus strains in Europe. The epidemiological implications of this finding for hantavirus infection in Europe and elsewhere are discussed. PMID- 9870583 TI - Primary and challenge infection of mice with equine herpesvirus 1, strain HSV25A. AB - Clinical signs, haematology, lymphocyte subset analysis, viral clearance, lung histopathology and humoral and cell-mediated (CMI) immune responses were monitored throughout the acute and convalescent phases of infection in groups of BALB/c mice infected intranasally with equine herpesvirus 1 (EHV-1), strain HSV25A. Primary infection caused a leucocytosis due to a neutrophilia during days 1 and 2 post-infection (pi) and a B lymphocytosis at day 1 pi. Serum ELISA antibodies were detected by 7 days pi and neutralising antibodies by 2 weeks pi. Mice infected with EHV-1 were not protected against disease when challenged with EHV-1 12 weeks later. However, viral clearance from lungs was significantly faster and the antibody response was markedly enhanced within the first few days of challenge infection. A CMI response was detected by 5 days after primary infection, but the level of responsiveness was not increased by challenge infection, although the lungs of challenged mice had markedly increased numbers of mononuclear cells around blood vessels and bronchioles. Specific antibodies to glycoprotein (g) B were detected by 2 weeks pi, 4 weeks earlier than the detection of antibodies to gC and 10 weeks before those to gD. The primary response was relatively short-lived with neither ELISA antibody nor lymphocyte proliferation was evident by 6 months pi. PMID- 9870585 TI - Monoclonal antibodies to the E2 protein of a new genotype (type 2) of bovine viral diarrhea virus define three antigenic domains involved in neutralization. AB - Bovine viral diarrhea virus (BVDV) has recently been segregated into two genotypes, namely, BVDV 1 and BVDV 2. Viruses of the BVDV 2 genotype are a cause of hemorrhagic and acute fatal disease in cattle in the US and Canada. In this study, monoclonal antibodies (mAbs) to the newly described BVDV 2 were produced after immunization with virus or a combination of virus and E2 peptide. From an original panel of 17 mAbs, 13 mAbs were identified as E2-specific by reactivity with a BVDV 2 recombinant E2 protein expressed in insect cells. Nine E2 mAbs were observed to be virus-neutralizing. The E2 epitopes represented by the mAbs were found to be highly conserved among BVDV 2 isolates associated with hemorrhagic or severe disease in cattle. Except for one virus-neutralizing E2 mAb, the mAbs showed few or relatively weak cross-reactions with BVDV 1. Two non-neutralizing E2 mAbs were BVDV 2-specific. In contrast to BVDV 1 for which conserved neutralizing epitopes have been mapped in one immunodominant domain, the virus neutralizing E2 mAbs produced to BVDV 2 were found to bind to highly conserved epitopes in three antigenic domains. PMID- 9870584 TI - Molecular cloning, expression and immunological analysis of the capsid precursor polypeptide (P1) from swine vesicular disease virus. AB - Swine vesicular disease virus (SVDV) is the aetiological agent of a highly contagious viral disease of pigs, whose symptoms are indistinguishable from those caused by foot-and-mouth disease virus (FMDV). The gene coding for the capsid protein precursor of SVDV (P1) from a recent spanish isolate (SPA/1/'93) was cloned and expressed in bacteria, and the antigenicity and immunogenicity of the recombinant product were evaluated. The recombinant P1 was recognised by antibodies against SVDV induced in pigs infected experimentally with different SVDV strains. Immunisation of swine with recombinant P1-induced SVDV-specific cellular and humoral immune responses. The implications of these results in SVD diagnostic as well as in vaccine development are discussed. PMID- 9870586 TI - Susceptibility to recombination rearrangements of a chimeric plum pox potyvirus genome after insertion of a foreign gene. AB - Infectious RNA transcripts were generated from a chimeric cDNA clone of the plum pox potyvirus (PPV) genome containing the bacterial beta-glucuronidase (GUS) gene inserted between the sequences coding for the P1 and HC proteins. An artificial cleavage site specific for the NIa viral proteinase was engineered between the GUS and HC sequences to produce free GUS and HC proteins. The resulting virus PPVGus/ was stably maintained during the first round of infection, although plants remained symptomless and virus accumulation was delayed with respect to wild-type infection. PPVGus/ deleted variants, missing between 645 and 1779 nt, were detected in a subsequent plant passage. PPVGus/ deletions were confined inside the GUS gene, never affecting the P1 and HC coding regions, in contrast with previous reports of deletions in other potyvirus-based vector, in which deletions frequently reached the HC gene. These results suggest that the N terminus of the PPV HC protein may be essential for virus viability. Analysis of the deletion endpoints showed short stretches of similarity in donor and acceptor RNAs, as well as oligo A tracts conserved in most junction sites, suggesting that deletions in PPVGus/ might take place by similarity-assisted recombination events. PMID- 9870587 TI - Evidence against KSHV infection in the pathogenesis of multiple myeloma. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is likely to play a pathogenic role in Kaposi's sarcoma, body cavity-based primary effusion lymphoma and a subset of Castleman's disease. A recent polymerase chain reaction (PCR)-based study reported an association between KSHV and multiple myeloma (MM). We searched for KSHV infection in MM patients by serology, PCR and immunohistochemistry. In addition, we cultured dendritic and stromal cells from MM patients. KSHV antibodies were universally absent from MM patients (0/25) whereas EBV antibodies were nearly ubiquitous (24/25). All of the bone marrow biopsies (0/16) and negative controls (0/4) were vIL-6 negative. None of the bone marrow aspirates (0/6) or biopsies (0/3), peripheral blood mononuclear cells (0/8), mononuclear apheresis cells (0/5) or dendritic cell cultures (0/5) were positive by PCR. One of the MM stromal cell cultures (1/7) was positive for KSHV DNA by PCR and weakly positive on direct southern hybridization using a probe to the terminal repeat region. However, this same patient was PCR negative using another primer set, KSHV seronegative, and negative for vIL-6 immunostaining. Our results suggest that the KSHV DNA positivity rate among MM patients is much lower than previously reported. PMID- 9870588 TI - Quantification of human T-cell lymphotropic virus type 1 proviral load by quantitative competitive polymerase chain reaction. AB - The polymerase chain reaction (PCR) has been established as a highly sensitive technique for detection of viral DNA or RNA. However, due to inherent limitations of PCR the amount of amplified product often does not correlate with the initial amount of template DNA. This is particularly true for PCR detection of viral infections that are characterized by low in vivo viral copy numbers in certain stages of the infection, such as human T-cell lymphotropic virus type 1 (HTLV-1) and simian T-cell lymphotropic virus type 1 (STLV-1). Therefore, we developed a quantitative competitive polymerase chain reaction (qcPCR) for detection of HTLV 1 and STLV-1 proviral DNA. The assay was optimized using an infectious HTLV-1 clone, ACH, HTLV-1 infected cell lines, MT-2.6 and HUT-102 and STLV-1 infected lines Kia and Matsu. Applicability of this system was demonstrated by determining HTLV-1 proviral load in peripheral blood mononuclear cells (PBMC) of human subjects with HTLV-1 associated diseases and an asymptomatic carrier as well as rabbits infected experimentally. This qcPCR method, the first designed specifically for HTLV-1 and STLV-1, will provide an important tool for pathogenesis studies of HTLV-1 and for evaluating the efficacy of antiviral drugs and vaccines against the viral infection using animal models. PMID- 9870589 TI - Application of a computer program for genetic typing of classical swine fever virus isolates from Germany. AB - The commercial software program HLA SequiTyper (Amersham Pharmacia Biotech), designed originally for human leukocyte antigen typing, was adapted for rapid typing of classical swine fever (CSF) virus isolates. The program compares new sequence data with those stored in a database file and calculates the most probable assignment. For generating the CSF virus sequence database, 150 bp of the 5' nontranslated genomic region (5'-NTR) from 88 German classical swine fever virus isolates from outbreaks between 1984 and 1997 were solid-phase sequenced directly after RT-PCR amplification. Sequence alignments showed that they all belonged to the previously defined genetic group 2. Within this group, six different subgroups could be distinguished, and were designated according to the geographic location where they are either still endemic or where they appeared most commonly. The advantage of using the HLA SequiTyper program is that it reads directly the sequence files as generated by the ALF sequencer (Amersham Pharmacia Biotech), making any manipulations unnecessary. In addition, a constant quality control of the raw sequence data can be achieved, as more than one sequence from the same isolate can be evaluated at once. Using this approach, new CSF isolates can be typed within 2 days. PMID- 9870590 TI - Expression of the enhanced green fluorescent protein by herpes simplex virus type 1 (HSV-1) as an in vitro or in vivo marker for virus entry and replication. AB - Glycoprotein K (gK) is involved in membrane fusion phenomena during infectious virus production and egress and is an important determinant for neurovirulence. To assess better the in vitro and in vivo roles of gK in virus replication, a recombinant virus was constructed expressing an engineered enhanced green fluorescent protein (EGFP) under the control of the human cytomegalovirus immediate early gene promoter (HCMV-IEP) inserted in place of the gK gene. The EGFP gene insertion was confirmed by diagnostic polymerase chain reaction (PCR), and the presence of the EGFP protein was detected by western immunoblot analysis using anti-GFP monoclonal antibody. Fluorescence microscopy revealed that virus infected cells emitted bright fluorescence when examined using filters for fluorescein. Fluorescence emission was detected as early as 4 h post-infection. Fluorescence intensity increased over time and was stable at late times after infection at which point viral plaques continued to emit bright green fluorescence. The amount of fluorescence emitted by virus infected Vero cells was monitored by fluorescence cytometry using a FACS cytometer. At an MOI of 3, all infected cells emitted strong green fluorescence as quantified by cytometry at 48 h post-infection. The deltagK-EGFP expressing recombinant virus will enable the determination of the role of gK in virus entry and egress as well as the role of gK in the molecular pathogenesis of herpes simplex virus type 1 (HSV-1). PMID- 9870591 TI - High-titer MSCV-based retrovirus generated in the pCL acute virus packaging system confers sustained gene expression in vivo. AB - Retroviral gene transfer using vectors encoding tumor suppressor genes has been tested repeatedly as a potential anti-tumor therapy. However, most attempts have been hindered by the inability to deliver genes efficiently and to obtain sustained expression in cells growing in vivo. In this paper we describe a method for producing high-titer MSCV virus using the pCL acute retroviral packaging system. This method facilitates the generation of MSCV virus encoding genes that convey the cytostatic or cytocidal phenotypes of benefit in the treatment of cancer. Amphotropic MSCV virus with an average titer of 6 x 10(6) CFU/ml has been routinely produced in this system. We demonstrate that, unlike the pCL retroviral vectors, the MSCV vector is capable of directing sustained in vivo expression of the green fluorescent protein in infected glioma cells following implantation and tumor growth in nude mice. PMID- 9870592 TI - Sensitivity of the Western blot detection of prion protein PrPres in natural sheep scrapie. AB - The sensitivity of Western blot detection of PrPres using two different extraction procedures on brain material of 30 scrapie-affected sheep was compared. Whereas PrPres could be detected in all sheep after extraction with the first method, 30% did not give any signal after extraction with the second method. However, the second method, when positive, permitted the detection of PrPres from smaller amounts of infected brain tissue. When used with the ruminant specific monoclonal antibody p4, the second method gave positive signals corresponding to less than 12.5 microg of scrapie-infected brain, that, up to now, is the highest sensitivity described for PrPres detection from naturally infected ruminant brains. The overall results showed highly variable levels of PrPres between sheep and are presented in relation to breed, survival time and animal genotype data. Further progress can thus be expected for PrPres detection in prion diseases, if more efficient extraction procedures and more sensitive immunological reagents are used. Such technical improvements could contribute to more accurate diagnosis in animals affected naturally. PMID- 9870593 TI - PCR sequencing of the spike genes of geographically and chronologically distinct human coronaviruses 229E. AB - A reverse transcription nested PCR (RT-PCR) sequencing methodology was developed and used to generate sequence data from the spike genes of three geographically and chronologically distinct human coronaviruses 229E. These three coronaviruses were isolated originally from the USA in the 1960s (human coronavirus 229E strain ATCC VR-74), the UK in the 1990s (human coronavirus 229E LRI 281) and Ghana (human coronavirus 229E A162). Upon translation and alignment with the published spike protein sequence of human coronavirus 229E 'LP' (isolated in the UK in the 1970s), it was found that variation within the translated protein sequences was rather limited. In particular, minimal variation was observed between the translated spike protein sequence of human coronaviruses 229E LP and ATCC VR-74 (1/1012 amino acid differences), whilst most variation was observed between the translated spike protein sequence of human coronaviruses 229E LP and A162 (47/1012 amino acid changes). Further, the translated spike protein sequence of human coronavirus 229E A162 showed three clusters of amino acid changes, situated within the 5' half of the translated spike protein sequence. PMID- 9870594 TI - Improvement of the print-capture polymerase chain reaction procedure for efficient amplification of DNA virus genomes from plants and insect vectors. AB - A rapid and simple procedure is described to amplify efficiently geminivirus DNA genomes by improving the print-capture polymerase chain reaction (PCR) procedure reported recently for RNA viruses. This method, termed print-PCR (P-PCR), allows direct amplification of DNA from infected plant or whitefly tissues printed directly on Whatman 3MM paper, without the need of any grinding, incubation, or washing steps previous to the amplification reaction. P-PCR reduces sample manipulation and avoids previous extraction of nucleic acids, thereby diminishing the possibilities of cross-contamination between samples. P-PCR has been successfully applied to whiteflies and various plant species infected by two different tomato yellow leaf curl viruses, TYLCV-Sr and TYLCV-Is, and for the amplification of the full-length genome of TYLCV-Is from infected plants. PMID- 9870595 TI - Electrophoretic analysis of the ribonucleoproteins of hepatitis delta virus. AB - Replication of hepatitis delta virus (HDV) is dependent on delta antigen (deltaAg), an HDV-encoded protein, which binds to HDV RNA and is capable of multimerization. To characterize HDV-specific ribonucleoprotein complexes (RNP) we used electrophoresis into non-denaturing agarose gels followed by northern analysis, to detect HDV RNA, and immunoblot, to detect deltaAg. We studied RNP from three sources: (i) vRNP, disrupted virions obtained from infected woodchuck serum; (ii) sRNP, disrupted particles secreted from transfected cultured cells; and (iii) cRNP, isolated from cells in which HDV genome replication was occurring. sRNP were approximately 28% smaller than vRNP. Treatment of vRNP with aurin tricarboxylic acid disrupted both deltaAg-deltaAg and deltaAg-RNA interactions while vanadyl ribonucleosides released the RNA without causing detectable disruption of the multimeric deltaAg complex. cRNP were smaller and more heterogeneous than vRNP and sRNP, and probably contained host components. The application of these electrophoretic procedures, and especially the use of prior treatments with vanadyl ribonucleoside complexes have provided valuable information on the RNP of HDV, and we expect they should find applicability in RNP studies of other RNA viruses. PMID- 9870596 TI - Prediction and a stationary, structured visual background influence the dynamics of the smooth-pursuit offset in humans. AB - It is still not clear whether the transition from pursuit eye movements to fixation is mediated by the same system that initiates pursuit, or whether another system, a specialized fixation system, is responsible. To investigate this question we measured smooth-pursuit eye movements and smooth-pursuit termination in five normal subjects using both predictable and unpredictable step ramp stimuli (velocities 10 degrees and 20 degrees/s) in front of a homogeneous and a structured visual background in order to compare the profile of eye velocity under these different conditions. With the predictable and/or structured visual background there was a gradual transition of eye velocity toward zero. In contrast, with the unpredictable stimulus in front of a homogeneous background, eye velocity during the offset was characterized by an overshoot (on the average, 2.2+/-1.0 degrees/s for 10 degrees/s ramps) before eye velocity settled at zero. Under this condition, steady-state velocity gain and the deceleration of the offset were significantly higher than during the other paradigm with the same target velocity. The latency of the pursuit offset was significantly shorter when a predictable stimulus was used. The duration of the offset did not depend on the experimental condition used. These findings imply that the pursuit onset and offset have some similarities and may be mediated by the same oculomotor system. PMID- 9870598 TI - Anteroposterior dynamic balance reactions to circular movement of the visual field without ocular pursuit. AB - It has been hypothesised that during natural vision, retinal image motion and eye motion information exert antagonistic effects on balance that cancel each other out. In a previous study, we found that pursuit movements of the eye unaccompanied by background slip on the retina induced strong, stereotyped anteroposterior sway in dynamic balance. In the present work, we investigated the influence of background retinal image slip alone on anteroposterior dynamic balance. We expected that retinal slip without ocular pursuit would induce large anteroposterior sway. Circular slip without rotation of the whole visual field image on the retina was obtained using a rotating prism. In this study, carried out on monocular vision, eye movement was prevented by fixating a stationary target seen through a small aperture in the prism. Anteroposterior dynamic balance conditions were elicited by a rocking platform. Strong, stereotyped anteroposterior sway accompanied by a visual illusion of target motion was observed. The induced sway was synchronised to the stimulus and led to a significant disturbance of balance. The similarity in magnitude of the effects of both retinal image motion studied here and eye motion information obtained previously, and their opposing phases, strongly support the initial hypothesis that the antagonistic effects of the two sources of information provided by normal ocular pursuit could interact in an additive algebraic mode. PMID- 9870597 TI - The distribution of primary afferent terminals from the eyelids of macaque monkeys. AB - Trigeminal sensory afferents from the eyelids convey two types of information that are important for the blink reflex. Movement of the lashes activates low threshold mechanoreceptors which evoke protective blinks. Information about eyelid position is also transmitted centrally and is used to adapt the metrics of the blink reflex to changing conditions over time. This study employed transganglionic transport of horseradish peroxidase conjugated to choleratoxin-B subunit or wheat-germ agglutinin to investigate trigeminal afferents supplying the eyelids of macaque monkeys. Ganglion cells labeled from upper- and lower-lid injections were located in the ophthalmic and maxillary portions of the trigeminal ganglion, respectively. In both cases, labeled terminals were observed ipsilateral to the injected eyelid in the principal and spinal trigeminal nuclei. However, only a few labeled terminals were present in the principal nucleus, and very sparse terminal labeling was confined to a few locations along the ventral border of the pars oralis and interpolaris of the spinal trigeminal nucleus. The main concentration of label was found in the pars caudalis at and immediately below the spinomedullary junction. The terminal field from the upper eyelid was located ventrally in the pars caudalis, and that from the lower eyelid was located more dorsally. In both cases, the labeled terminal field was densest within lamina II of the spinal trigeminal nucleus. The heavy concentration of eyelid central terminals at the spinomedullary junction is surprising in light of physiological studies indicating representation of all parts of the face throughout the trigeminal nucleus. The distribution of eyelid afferent terminals in the macaque is caudal to the main concentration of corneal afferent terminals at the pars interpolaris/caudalis border. This may be a basis for differences seen in blinks produced by corneal as opposed to supraorbital stimulation. The presence of a single major site of eyelid primary afferent terminals suggests that sensory input for both eyelid proprioception and blink-reflex activation passes through this segment of the spinal trigeminal nucleus. These results provide a basis for investigation of the central connections of pars caudalis neurons in order to better establish the pathways producing trigeminally evoked blinks and blink adaptation. PMID- 9870599 TI - Microstimulation of movements from cerebellar-receiving, but not pallidal receiving areas of the macaque thalamus under ketamine anaesthesia. AB - The motor thalamic areas receiving input from the globus pallidus (VA) and the cerebellar nuclei (VL) appear to have different roles in the generation and guidance of movements. In order to further test these differences, we used electrical stimulation to map the ventro-anterior and ventro-lateral nuclei of the thalamus in three ketamine anaesthetised monkeys. Movements were readily evoked from VL at currents of down to 10 microA. The movements were typically multijoint, and stimulation could evoke arm and trunk or arm and facial movement at the same current threshold. Evoked arm movements often involved multiple joints, with or without finger movements. Facial movements included the lips, tongue, jaw, eyebrows and, occasionally, the eyes. The thalamic map was topographic, but complex with at least two separate regions related to arm movement. Very few sites within the VA could stimulate movement, even at high currents. We therefore suggest that the cerebellar projections to motor regions of the cortex, which pass through the VL thalamic nuclei, have a different relationship and are closer to movement execution than the projections from basal ganglia via the ventro-anterior nucleus. PMID- 9870600 TI - An investigation of the intrinsic circuitry of the motor cortex of the monkey using intra-cortical microstimulation. AB - The motor cortex contains a distributed map of muscles, with a single muscle represented over a wide cortical area. We have searched for inter-connections between distant sites projecting to common muscles by delivering pairs of 20 microA single-pulse intracortical microstimuli (ICMS) to sites separated by 1.5-2 mm in the hand-area primary motor cortex of two macaque monkeys performing a precision grip task. The facilitation of hand- and forearm-muscle rectified EMG was measured. When stimuli were delivered simultaneously, responses were quantified using a technique to correct for non-linearities inherent in the use of averaged, rectified EMG. A spatial facilitation was seen for such simultaneous stimuli; however, it was of the same magnitude as that occurring when ICMS was paired with stimulation of corticospinal axons in the pyramidal tract (PT), so that it was likely to be spinal in origin. When two such distant sites were stimulated separated by a 10- or 20-ms delay, the second response scaled with the level of background EMG in the same way as a response to the PT stimulus. By contrast, when the same site was stimulated twice with these delays, the second response showed a facilitation compared with a similarly timed PT response. There would therefore appear to be a local facilitation of the cortical output at these intervals, which is not seen between distant sites. Antidromically identified pyramidal-tract neurones (PTNs) were recorded whilst stimuli were delivered to a cortical site, with a distance between stimulating and recording electrodes of also 1.5-2 mm. The most common response was a facilitation followed by a suppression. Six of eleven PTNs showed a facilitation in their discharge following this stimulation (maximum connection strength s=0.19), 8/11 showed a suppression (maximum s=0.16). It is concluded that powerful inter-connections do exist between distributed parts of the motor output and that there is widespread cortical activation after even a single ICMS pulse. However, these inter connections do not lead to interactions between cortical outputs following stimulation, as assessed from the EMG. It is proposed that this is likely to reflect differences in the summation of output cells to local versus remote stimulation. PMID- 9870601 TI - Dynamics of intergestural timing: a perturbation study of lip-larynx coordination. AB - In this study, downward-directed mechanical perturbations were applied to the lower lip during both repetitive (/...paepaepae.../) and discrete (/pe'saepaepl/) utterances in order to examine the perturbation-induced changes of intergestural timing between syllables (i.e., between the bilabial and laryngeal gestures for successive /p/'s) and within phonemes (i.e., between the bilabial and laryngeal gestures within single /p/'s). Our findings led us to several conclusions. First, steady-state (phase-resetting) analyses of the repetitive utterances indicated both that "permanent" phase shifts existed for both the lips and the larynx after the system returned to its pre-perturbation rhythm and that smaller steady-state shifts occurred in the relative phasing of these gestures. These results support the hypothesis that central intergestural dynamics can be reset by peripheral articulatory events. Such resetting was strongest when the perturbation was delivered within a "sensitive phase" of the cycle, during which the downwardly directed lower-lip perturbation opposed the just-initiated, actively controlled bilabial closing gesture for /p/. Although changes in syllable duration were found for other perturbed phases, these changes were simply transient effects and did not indicate a resetting of the central "clock." Second, analyses of the transient portions of the perturbed cycles of the repetitive utterances indicated that the perturbation-induced steady-state phase shifts are almost totally attributable to changes occurring during the first two perturbed cycles. Finally, the transient changes in speech timing induced by perturbations in the discrete sequences appeared to share a common dynamical basis with the changes to the repetitive sequences. We conclude by speculating on the type of dynamical system that could generate these temporal patterns. PMID- 9870602 TI - Off-center yaw rotation: effect of naso-occipital linear acceleration on the nystagmus response of normal human subjects and patients after unilateral vestibular loss. AB - Dual search coils were used to record horizontal, vertical and torsional eye movement components of one eye during nystagmus caused by off-center yaw rotation (yaw centrifugation). Both normal healthy human subjects (n=7) and patients with only one functioning labyrinth (n=12) were studied in order to clarify how the concomitant linear acceleration affected the nystagmus response. Each subject was seated with head erect on the arm of a fixed-chair human centrifuge, 1 m away from the center of the rotation, and positioned to be facing along a radius; either towards (facing-in) or away from (facing-out) the center of rotation. Both yaw right and yaw left angular accelerations of 10 degrees s(-2) from 0 to 200 degrees/s were studied. During rotation a centripetal linear acceleration (increasing from 0 to 1.24xg units) was directed along the subject's naso occipital axis resulting in a shift of the resultant angle of the gravitoinertial acceleration (GIA) of 51 degrees in the subject's pitch plane and an increase in the total GIA magnitude from 1.0 to 1.59xg. In normal subjects during the angular acceleration off-center there were, in addition to the horizontal eye velocity components, torsional and vertical eye velocities present. The magnitude of these additional components, although small, was larger than observed during similar experiments with on-center angular acceleration (Haslwanter et al. 1996), and the change in these components is attributed to the additional effect of the linear acceleration stimulation. In the pitch plane the average size of the shift of the axis of eye velocity (AEV) during the acceleration was about 8 degrees for a 51 degrees shift of the GIA (around 16% of the GIA shift) so that the AEV-GIA alignment was inadequate. There was a very marked difference in the size of the AEV shift depending on whether the person was facing-in [AEV shift forward (i.e. non-compensatory) of about 4 degrees] or facing-out [AEV shift forward (i.e. compensatory) of around 12 degrees]. The linear acceleration decreased the time constant of decay of the horizontal component of the post-rotatory nystagmus: from an average of 24.8 degrees/s facing-in to an average of 11.3 degrees/s facing-out. The linear acceleration dumps torsional eye velocity in an manner analogous to, but independent of, the dumping of horizontal eye velocity. Patients with UVD had dramatically reduced torsional eye velocities for both facing-in and facing-out headings, and there was little if any shift of the AEV in UVD patients. The relatively small effects of linear acceleration on human canal-induced nystagmus found here confirms other recent studies in humans (Fetter et al. 1996) in contrast to evidence from monkeys and emphasizes the large and important differences between humans and monkeys in otolith-canal interaction. Our results confirm the vestibular control of the axis of eye velocity of humans is essentially head-referenced whereas in monkeys that control is essentially space-referenced. PMID- 9870603 TI - Effects of excitotoxic lesions of the rat ventral striatum on the perception of reward cost. AB - This study examined the possibility that lesions of the nucleus accumbens in rats impair the perception of the "cost of reward", as defined by the number of operant responses needed to obtain a food pellet. In a first experiment, visual cues indicated the cost of reward under a multiple-ratio schedule of reinforcement. In a second experiment, the number of lever presses required for each reward incremented with each trial in a progressive-ratio schedule of reinforcement. Lesions of the nucleus accumbens altered the behavioral response to the increasing cost of reward when there was an absence of external cues. There was no change in the ability of the lesioned rats to respond to visual cues that indicated reward availability. The results are considered in terms of the traditional idea of the nucleus accumbens as a limbic-motor interface: it is suggested that, if the nucleus accumbens serves such a function, it is limited to only some contexts. PMID- 9870604 TI - Plasticity of tonotopic maps in auditory midbrain following partial cochlear damage in the developing chinchilla. AB - There is substantial reorganization of the midbrain (inferior colliculus) tonotopic map following neonatally induced partial cochlear lesions in the chinchilla. The most obvious feature of this remapping is a large "iso-frequency" region in the ventral sector of the central nucleus of inferior colliculus (ICC). Neurons in this region exhibit similar threshold and tuning properties, with a common characteristic frequency which corresponds to the high-frequency audiometric cutoff. This overrepresented frequency range also corresponds to the high-frequency border of the cochlear lesion. Alterations to the tonotopic map corresponding to lower frequencies, in more dorsal regions of ICC, depend on the extent and degree of the cochlear lesion. When there is minimal damage to apical (low-frequency) cochlear areas, the dorsal ICC has relatively normal frequency representations. With more extensive apical cochlear lesions there is a corresponding disruption of ICC tonotopic representation of the low frequencies. We conclude that the tonotopic map within the ICC can become (re)organized postnatally according to the abnormal pattern of neural activity from the auditory periphery. Similar reorganization can be expected to occur in human infants with a partial cochlear hearing loss from birth. PMID- 9870605 TI - Movement-related potentials with reference to isometric force output in discrete and repetitive tasks. AB - This study investigates whether different speed and accuracy constraints in discrete and repetitive index finger isometric force-production tasks influence the characteristics of the movement-related potentials (MRP) preceding and accompanying these tasks. Three components of MRP (Bereitschaftspotential, BP, motor potential, MP, and movement-monitoring potential, MMP) associated with isometric force output were identified and examined. Our principal finding for the MRP amplitude showed that only MMP, not BP and MP, was enhanced at higher rates of force development for both speed and accuracy tasks. That is to say, there was a high correlation between MMP peak amplitude and the rate of force development for both repetitive and discrete force-production tasks. Additionally, the amplitude of MMP was consistently higher for fast, rather than accurate, force outputs. Moreover, the results from analysis of MRP onset times suggest that, in general, the MRP begin earlier for the fast force output than for the accurate force output. PMID- 9870606 TI - Simple artificial neural network models can generate basic muscle activity patterns for human locomotion at different speeds. AB - A neural network model has been developed to represent the shaping function of a central pattern generator (CPG) for human locomotion. The model was based on cadence and electromyographic data obtained from a single human subject who walked on a treadmill. The only input to the model was the fundamental timing of the gait cycle (stride rate) in the form of sine and cosine waveforms whose period was equal to the stride duration. These simple signals were then shaped into the respective muscle activation patterns of eight muscles of the lower limb and trunk. A network with a relatively small number of hidden units trained with back-propagation was able to produce an excellent representation of both the amplitude and timing characteristics of the EMGs over a range of walking speeds. The results are further discussed with respect to the dependence of some muscles upon sensory feedback and other inputs not explicitly presented to the model. PMID- 9870607 TI - Electrophysiological localization of brain regions involved in perceptual memory. AB - Event-related potentials (ERP) were recorded during perceptual discrimination and short-term memory, varying the interstimulus interval (1-10 s) in delayed spatial frequency discrimination. Accuracy of discrimination remained unimpaired across this time interval, but choice reaction times increased. A brain source localization (BESA) model showed that the activity of the parietal and right temporal sources increased with long retention intervals in a sequential activation pattern where a long-latency component of the parietal source specific to the memory condition was observed, the latency of which matched a memory related increase in choice reaction times in the cognitive task. It is suggested that the temporal sources are involved in encoding and storage of visual information, and the parietal source is involved in memory retrieval. PMID- 9870608 TI - Cell surface GPI-anchoring of CD45 isoforms. AB - We have designed a new cell surface expression plasmid to study the structural and membrane-topological requirements for functioning of different isoforms of CD45, a leucocyte specific member of the protein tyrosine phosphatase (PTPase) family of proteins. Use of this vector in cell transfection experiments enabled us to produce multiple CD45 isoforms (ABC, B, Null), with their extracellular segment intact, and the entire membrane spanning and intracellular C-terminal domain replaced by a GPI-membrane-anchor and VSV-tag. Our strategy facilitated the identification and analysis of chimeric proteins and selection of cell clones from low transfection efficiency experiments. We demonstrate here that simple expression of GPI-anchored CD45 isoforms on transfected Cos-1 cells does not facilitate binding to CD22+ lymphoid cells. This suggests that not only the mere presence of CD45 extracellular domains but also their assembly into higher order structures at the cell surface, is necessary in order to engage in the recognition and/or signalling processes normally used by B- and T-cells. PMID- 9870609 TI - A rearrangement of the CDD gene at the 5' UTR produces two types of transcripts that contain a natural antisense region. AB - In order to study the regulatory region for transcription, the genomic DNA of the human CDD gene was cloned and analyzed. In contrast to previously reported CDD cDNA sequence, the sequence of the isolated genomic clone was rearranged at the 5' untranslated region (UTR) by a 30 bp inversion and a 57 bp insertion. Polymerase chain reaction (PCR) of chromosomal DNA and mRNA, and sequencing analysis of the PCR products revealed that sequences corresponding to both the genomic clone and the cDNA clone were present in the chromosomal DNA and were also transcribed into mRNA. Because of the 30 base inversion, the two types of CDD transcripts contain antisense sequences in their 5' UTR. Their role in the regulation of CDD expression is discussed. PMID- 9870610 TI - The dual identities of mammalian tRNA(Sec) for SerRS and selenocysteine synthase. AB - Se is an essential trace element and is found as a selenocysteine in the active site of Se-enzymes, such as glutathione peroxidase. tRNASec is first aminoacylated with serine by Ser RS and further is converted to selenocysteyl tRNA by selenocysteine synthase. Mammalian selenocysteine tRNA has dual identities with Ser RS and selenocysteine synthase. Key identity elements for selenocysteine synthase are the long 9 bp AA- and long 6 bp D-stems. Major serine tRNA was converted to a mutant with a 9 bp AA-stem and 6 bp D-stem, instead of a 7 bp AA-stem and 3 bp D-stem. This mutant was active for selenylation as well as serylation. The relative kinetic parameter (Vmax/Km) of the mutant was 0.052 of the value (1.00) of wild-type Sec tRNA. This low value suggests that there is an unknown fine base specific for selenocysteine synthase. For serylation, mutant having 12 bp and wild type tRNASec having 13 bp of the total length of AA- + T stems were active but the mutants having 11 or 14 bp were inactive. This shows that SerRS measures the distance between the discrimination base and long extra arm for recognition of tRNASer. PMID- 9870611 TI - Detection of TNF alpha and Fas ligand mRNA within synovial mononuclear cells by fluorescence in-cell labeling PCR (FICL-PCR). AB - T cells that infiltrate the synovial lesions of rheumatoid arthritis may play a key role in its pathogenesis. To learn more about their functional nature, we determined the frequency of synovial T cells that harbored the TNF alpha and Fas ligand transcript by a technique, called Fluorescence In-Cell Labeling Polymerase Chain Reaction (FICL-PCR). The mRNA of interest was detected in fixed cells by the incorporation during PCR of a fluorescein-12-dUTP label following an initial reverse transcription PCR step. Using this technique the CD3 transcript was detected in the T leukemic cell line, MOLT-4, with calculated sensitivity and specificity values of 91% and 100%, respectively. The percentage mean (+/-S.D.) of TNF alpha mRNA positive cells and Fas ligand mRNA positive cells in peripheral blood mononuclear cells from 12 rheumatoid arthritis patients were 5.1+/-2.3% and 4.8+/-3.1%, respectively. The percentage mean (+/-S.D.) of TNF alpha mRNA positive cells and Fas ligand mRNA positive cells among synovial mononuclear cells from six rheumatoid arthritis patients was 16.8+/-8.3% and 10.8+/-1.8%, respectively. This result indicates that the cytotoxic T cells expressing TNF alpha accumulate in rheumatoid arthritic lesions where they may play a pathogenic role. PMID- 9870612 TI - The rodent alphaA-crystallin gene: mutagenesis of a non-consensus 5'-splice site to study alternative splicing in vivo. AB - alphaA-Crystallin is a member of the small heat shock protein family that is abundantly expressed as a structural component in the vertebrate eye lens. In lenses of rodents and some other mammals, there occurs a minor variant of alphaA crystallin, which has an insertion of 23 amino acid residues. This variant, alphaA(ins)-crystallin, results from differential integration of an optional exon into a small fraction of the mRNA. We have studied whether this alternative splicing is caused by a non-consensus cytosine in the 5' splice site adjacent to the optional exon. After replacement of the aberrant cytosine in the hamster alphaA-crystallin gene by a consensus thymine, and transient transfection of this gene in Chinese Hamster Ovary cells, the optional exon is indeed almost completely spliced into the mature mRNA. In contrast, replacement of the cytosine by adenine or guanine completely abolishes the splicing of the optional exon. Our results confirm that alternative splicing of the alphaA-crystallin primary transcript is mainly due to a non-consensus 5' splice site nucleotide. However, we conclude that the small size of the optional exon is probably an additional contributing factor and therefore it seems that the splicing mechanism is based on recognition of exons rather than introns. PMID- 9870613 TI - Transactivation by expression of the hepatitis B virus X protein with an inducible system. AB - We describe here the conditional expression of the hepatitis B virus X protein using the inducible system controlled by a tet-responsive promoter. Induction of the X protein in Rat-2 fibroblasts activated transcription from a heterologous gene promoter and stimulated the DNA-binding activity of NFkB. The ability to produce this biologically active X protein in a stable cell line will accelerate the elucidation of the function and mechanisms of X and eventually help us understand the role of X in natural viral infection and carcinogenesis. PMID- 9870614 TI - Effects of polyamines on higher-order folding of in situ chromatin. AB - Modifications of the higher-order chromatin structure induced by polyamines have been quantitatively investigated in situ through a non-invasive biophysical approach using Differential Scanning Calorimetry and Quantitative Fluorescence Microscopy. Calorimetric and intensitometric profiles have been acquired for samples of native thymocytes, alternatively suspended in buffers, with or without natural polyamines (spermine and spermidine). The results here reported show that the structure and distribution of nuclear chromatin in situ considerably change upon the ionic composition of the environment. A quantitative analysis of this data and a comparison with previous results obtained from isolated chromatin fibers was carried out. Finally, an inverse relationship between chromatin condensation and nuclear volume was observed. PMID- 9870615 TI - Activation of vitellogenin II gene expression by steroid hormones in the old Japanese quail. AB - Alterations in the basal transcription rates of eukaryotic genes are believed to involve the binding of trans-acting factor(s) with specific DNA sequences in the promoter. We show here two interrelated events for the VTGII gene of the old, non egg laying Japanese quail: alterations in the structure of the chromatin encompassing the gene, and binding of trans-acting factors to the promoter of the gene. Estradiol/progesterone alone or together cause alterations in the conformation of the chromatin of the promoter region of the gene. This may allow free access of nuclear protein(s) to the cis-acting elements, ERE, PRE and NF1, in the promoter of the gene and cause activation of transcription. PMID- 9870616 TI - The different alternatively spliced isoforms of the Oct-2 transcription factor repress the involucrin promoter in a cell type-specific manner. AB - It has previously been reported that several octamer binding transcription factors including the Oct-2 factor can repress the involucrin gene promoter in keratinocyte cells. As the Oct-2 factor exists in several different cell type specific isoforms with distinct activating or inhibiting effects on gene expression, we have tested the effect of these forms on the involucrin promoter. We report here that at high concentrations each of these isoforms can inhibit the involucrin promoter in keratinocytes. In other cell types however, all three isoforms activate the involucrin promoter and this effect is also observed at low concentrations of Oct-2 in keratinocytes. The mechanisms responsible for these effects are discussed. PMID- 9870617 TI - The non-specific effect of endolymphatic sac surgery in treatment of Meniere's disease: a prospective, randomized controlled study comparing "classic" endolymphatic sac surgery with the insertion of a ventilating tube in the tympanic membrane. AB - A prospective, randomized study was carried out comparing the effect of two surgical modalities in the treatment of patients with Meniere's disease: insertion of an endolymphatic sac shunt and insertion of a ventilating tube in the tympanic membrane. A total of 29 patients, 12 males and 17 females, age 27-71 years, were operated on in two ear, nose and throat (ENT) departments. Of these patients, 15 had an endolymphatic shunt inserted and 14 had a ventilating tube inserted in the tympanic membrane. Postoperative follow-up was carried out in the department in which the patients had not been operated. The severity of the disease was scored pre- and postoperatively, and the results evaluated under the guidelines of the Committee on Hearing and Equilibrium (1995) for the diagnosis and evaluation of therapy in Meniere's disease. The patients in both groups had a statistically significant reduction in dizzy spells, measured 6 and 12 months postoperatively, and there was no difference between the groups. The pathophysiological explanation for the reduction in dizzy spells in each of the treatment modalities is debatable and the effect is non-specific. The patients' hearing and tinnitus were statistically unaffected by the treatment in both groups, though 2 patients in the shunt group developed severe hearing loss (anacusis/70 dB). PMID- 9870618 TI - The familial incidence of benign paroxysmal positional vertigo. AB - The goal of this investigation was to determine whether there is a familial tendency in the development of benign paroxysmal positional vertigo (BPPV). We hypothesized an increased frequency of BPPV among relatives of patients with the same diagnosis. BPPV is caused by dislodged otoconia from the utricular macula floating in the semicircular canals. At least half of BPPV cases are idiopathic and most pathological associations provide no clue as to the reason otoconia become dislodged. We have noted a number of BPPV patients with family histories of BPPV, suggesting a genetic predisposition to the condition. We surveyed 120 successive BPPV patients and 120 successive dizzy patients without BPPV regarding the frequency of dizziness and BPPV (diagnosed by a physician) among family members. Patients in our group with BPPV were 5 times as likely to have relatives with BPPV compared to the dizzy control group (chi2=5.95, DF=1, p=0.015). We have demonstrated that there is a familial tendency for the occurrence of BPPV. There is nothing in our data that would distinguish between a hereditary or environmental influence in the development of the disease, however. PMID- 9870619 TI - Passive sustained turning of the head induces asymmetric gain of the vestibulo ocular reflex in healthy subjects. AB - In order to test the hypothesis of an interaction between neck proprioception and the vestibulo-ocular reflex (VOR), we rotated 16 healthy subjects both facing forward and with their heads passively turned 70 degrees to either side. We found that gain tended to be lower when the subjects were rotated with their heads turned opposite to the direction of rotation compared to when they were rotated in the same direction, but facing forward. Although our findings were not statistically significant, they suggest that there is a measurable interaction between neck proprioception and the VOR in subjects with normal vestibular function. Asymmetric neck muscle proprioceptive signals seem to give rise to asymmetric functioning of the VOR, which, at least in part, could be the pathogenesis of cervical dizziness. If so, this could lead to misinterpretation of vestibular assessments in patients with neck pain who also complain of dizziness. PMID- 9870620 TI - House-Brackmann and Yanagihara grading scores in relation to electroneurographic results in the time course of Bell's palsy. AB - The results of House Brackmann and Yanagihara grading were compared with electroneurographic (ENoG) data in 30 consecutive patients with Bell's palsy. The examinations were made on mean days 11, 36 and 99. Twenty-four patients had a favourable outcome (Yanagihara > or = 36 at three months). Based on our observations, 23 (96%) of these could have been predicted by ENoG, 18 (75%) by Yanagihara grading and 6 (25%) by House Brackmann grading. Initially, the relative House Brackmann scores showed a slightly milder palsy than the Yanagihara scores, but in the follow-up period the gradings were almost identical. The mild palsies, defined on the initial ENoG results, initially demonstrated relatively less nerve dysfunction on ENoG than the clinical grading; by the first follow-up, the ENoG and clinical grading had both returned to normal. The intermediate palsies had almost the same initial relative clinical and ENoG values, but at the first follow-up (mean day 36), the facial function had returned to normal despite abnormally reduced, but improved, ENoG values. In the severely affected patients, the follow-up studies showed an improved clinical function but ENoG values still demonstrated a high degree of degeneration (slightly improved at second follow-up). In this study, patients with a favourable outcome were best predicted with ENoG. Clinical identification of these patients was more accurate with Yanagihara than with House Brackmann. Furthermore, in all three groups a clinical improvement, due to the release of neurapraxia, was noted at the first follow-up. The slow ENoG improvement noted at follow-up was probably due to nerve regeneration by collateral sprouting. Based on the time course of our ENoG findings, it appears that patients with a high degree of degeneration at both the initial examination and first follow-up have a poorer prognosis. PMID- 9870621 TI - Significance of trigeminal sensory input on regrowth of hypoglossal and facial motoneurons after hypoglossal facial anastomosis in rats. AB - Hypoglossal facial anastomosis (HFA) is a standard surgical technique for restoration of facial movements in cases of intratemporal lesions of the facial nerve. Case reports provide evidence that an affected trigeminal system reduces functional outcome. In order to detect morphological changes in the hypoglossal nucleus responsible for this phenomenon, we used 18 Wistar rats and performed three different surgical combinations. In group 1, six animals received HFA only. In group 2, HFA was combined with resection of the contralateral infraorbital nerve. In group 3, HFA was combined with resection of the ipsilateral infraorbital nerve. Fifty-six days after the operation, horseradish peroxidase (HRP) was injected into the whisker pad. As shown in previous studies using HRP, retrograde-labelled motoneurons occurred in the hypoglossal and facial nuclei. Counts of the labelled motoneurons showed no change in the number of projecting hypoglossal motoneurons in group 2 when compared to HFA only, but a significantly smaller number in group 3 (-35%). Furthermore, the number of projecting facial motoneurons was significantly reduced in group 2 (-85%) and group 3 (-45%). These morphological findings indicate an absent or insufficient functional connection between the contralateral infraorbital nerve and the hypoglossal nucleus, and a strong influence of the infraorbital nerve to the ipsi- and contralateral facial nuclei. Additionally, our study provides morphological evidence that the integrity of the sensory trigeminal system is very important in reconstructive facial nerve surgery. PMID- 9870622 TI - Influence of speech-coding strategy on cortical activity in cochlear implant users: a positron emission tomographic study. AB - The effects of a speech-coding strategy of cochlear implant (CI) on cortical activity were evaluated using positron emission tomography. The CIs used in the present study were those of a 22-channel system using the Multipeak speech-coding strategy (MPEAK) and the spectral peak strategy (SPEAK). On comparing the 2 groups, it was found that the speech-tracking performance was significantly higher in the SPEAK group than in the MPEAK group. Regional cerebral blood flow (rCBF) was measured during the silent resting, noise stimulus and speech stimulus periods. The increase in rCBF was localized mainly in the primary auditory area during the noise stimulus period. The increase in rCBF in the auditory association area during the speech stimulus period was stronger in the SPEAK group than in the MPEAK group. This finding suggests that the SPEAK strategy activates more speech processing neuronal networks in the auditory association area than the MPEAK strategy. PMID- 9870623 TI - The influence of an arterial and venous air embolism on the hearing level in laboratory mini pigs. AB - We examined the influence of an experimental venous and arterial air embolism on the hearing level in laboratory mini pigs. Before and after the injection of air a threshold ABR was measured in anaesthetized mini pigs (n=15). A venous air embolism was performed in 6 animals: no changes in the hearing level or in the interpeak latencies Jewett wave V-I were observed in any of the animals before, during or after the application of air. In 9 animals the arterial embolism was done, 2 animals died and had to be excluded. In 6 animals out of 7 a hearing loss was observed starting after the injection of 40 ml air and finally ending in deafness for these 6 animals. The detected hearing loss is probably of cochlear origin, as no prolongations of the interpeak latencies of Jewett wave V-I at 90 dB HL were observed prior to the deafness. Our results show that air bubbles in the arterial circulation lead to cochlear damage ending in deafness. Our research indicates that decompression sickness, which is comparable to the arterial air embolism, is more often the cause of a sensorineural hearing loss after diving than previously believed. PMID- 9870624 TI - Investigation of topical ciprofloxacin ototoxicity in guinea pigs. AB - Antibiotic eardrops mostly contain potentially ototoxic aminoglycosides. Ciprofloxacin is an alternative, and there is limited experience in its topical use. To investigate the topical ototoxicity of ciprofloxacin, 11 guinea pigs have been operated on. Transbullae silicone drug delivery tubes were placed to both ears of the animals. After the operation the guinea pigs were divided into two groups. The first group of animals received 0.2 ml of 4% gentamicin in one ear and 0.2 ml of 0.9% sodium chloride solution in the other. The second group received 0.2 ml of 0.2% ciprofloxacin in the test ear and 0.2 ml of 0.9% sodium chloride solution in the control ear. All drugs were given once a day on 7 consecutive days. Auditory brainstem response thresholds were recorded using click, 4 and 8 kHz logon stimuli before and after the operation, and after topical drug application. Results were statistically compared using Wilcoxon matched pairs signed-ranks test. Comparison of the thresholds before and after the operation, physiological saline application, as well as ciprofloxacin application yielded no statistically significant differences, whereas application of gentamicin resulted in total hearing loss. The results indicate that topical use of 0.2% ciprofloxacin is not ototoxic in guinea pigs. PMID- 9870625 TI - Effects of impulse noise on cortical response threshold and inner ear activity of succinic dehydrogenase and acetylcholinesterase in guinea pigs. AB - The effects of impulse noise (firecrackers at 170 dB SPL, 1, 10, 20 rounds) on auditory cortical response threshold (CRT) and activity of succinic dehydrogenase (SDH) and acetylcholinesterase (AchE) in the inner ear were studied in 37 guinea pigs. The results showed that extent of damage in the cochlea was related to amount of exposure to the noise. Exposure to 10 rounds resulted in temporal threshold shift (TTS); to 20 rounds the result was permanent threshold shift (PTS). For the period when TTS existed, inverse correlation was noticed between enzyme activity change and CRT shift. The correlation could not be established when PTS was induced. The results suggest that the pathomechanism of PTS was more complex than that of TTS. The significance of the results is discussed. PMID- 9870626 TI - Effect of norepinephrine on ouabain-sensitive, K+-dependent p nitrophenylphosphatase activity in strial marginal cells of the cochlea in normal and reserpinized guinea pigs. AB - On the basolateral infoldings of the strial marginal cells in the cochlea, Na K ATPase activity is abundant. To clarify the humoral control by norepinephrine, K NPPase activity of strial marginal cells in the cochlea was investigated in normal, reserpine, norepinephrine (NE), reserpine plus NE-treated guinea pigs using a cerium-based method. K-NPPase activity was almost completely decreased 3 20 days after reserpine administration. At 10 days after reserpinization and following NE repeated treatment, enzyme activity was detectable. These results suggested that norepinephrine might restore and regulate strial K-NPPase activity. PMID- 9870627 TI - Gap junction change in supporting cells of the organ of Corti with ryanodine and caffeine. AB - It has been demonstrated that the gap junctions of the supporting cells of the organ of Corti are controlled by H+ and Ca2+. Inside these cells there is a tubular structure. It is supposed that this network is endoplasmic reticulum. Calcium release from inside the cells, and the effect of calcium on the gap junctions of these cells, were investigated under whole cell clamping application of ryanodine and caffeine. Membrane capacitance and membrane resistance were calculated, with corrections for changes in whole cell parameters. Ryanodine treated cells (1 microM-10 mM), caffeine-treated cells (5 mM 500 nM) and A23187 treated cells were uncoupled at their gap junctions. Therefore, Ca2+ plays a role in the uncoupling of the gap junctions in supporting cells of the organ of Corti from inside the cells. PMID- 9870628 TI - Serum antibodies specific to CD outer membrane protein of Moraxella catarrhalis, P6 outer membrane protein of non-typeable Haemophilus influenzae and capsular polysaccharides of Streptococcus pneumoniae in children with otitis media with effusion. AB - We measured the levels of serum IgG antibodies to CD outer membrane protein of Moraxella catarrhalis, P6 outer membrane protein of non-typeable Haemophilus influenzae and capsular polysaccharides of Streptococcus pneumoniae in 168 children with otitis media with effusion (OME) who were followed prospectively, using ELISA. Serum IgG antibodies to CD, P6 and pneumococcal capsular polysaccharides were detected in all samples. The anti-pneumococcal polysaccharides antibody level was highest, followed by the anti-P6 antibody level and anti-CD antibody was lowest (median:interquartile ranges were 45.9:19.1 100 microg/ml, 15.6:9.70-23.2 microg/ml and 1.06:0.73-1.87 microg/ml, respectively). In children aged 0-6 years, there were positive correlations among the antibody levels (anti-CD vs anti-P6, r=0.325, p <0.001; anti-CD vs anti polysaccharide, r=0.397, p <0.0001; anti-P6 vs anti-polysaccharide, r=0.175, p=0.057). However, no relationship was seen in children aged 7-15 years. Children were classified according to severity of OME during the 1-year follow-up. In children aged 0-6 years, the severity of OME correlated inversely with the levels of anti-CD antibody (r=-.23, p=0.012), of anti-P6 antibody (r=-0.292, p=0.0015), and of anti-pneumococcal polysaccharides antibody (r=-0.25, p=0.0064). However, no correlation was found between antibody levels and severity of OME in children aged 7-15 years. These data suggest that persistence and/or recurrence of OME may be due to an insufficient serum antibody response to middle ear pathogens in young children. PMID- 9870629 TI - Congenital bony fixation of the malleus. AB - Bony fixation of the malleus seems to be one of the most controversial clinical entities among various congenital disorders of the auditory ossicles. According to various theories, it could be caused by trauma, chronic infection of the ear, otosclerosis or even development failures in the tympanic cavity itself. Histological analysis of 1,108 temporal bones showing a normal middle ear structure was performed. All bones were cut in serial sections of 20 microns. Audiograms and the data on ear trauma or chronic infection were analysed from case histories belonging to each bony specimen. Bony fixation of the malleus was found in 14 cases. It was almost always unilateral. The most frequent site of the fixation was the lateral epitympanal wall. The forms of fixation differed from a thin bony lamella to a solid bony bridge. The mallear ligaments were not involved in any case. No relationship to chronic ear infection, ear trauma or otosclerosis was found. Only a slight conductive hearing impairment up to 20 dB was recorded in 6 out of 14 cases. The most probable predisposing anatomical factors for the onset of the bony fixation of the malleus and the most recent preoperative diagnostic possibilities are discussed. PMID- 9870630 TI - Postnatal development of Eustachian tube: a computer-aided 3-D reconstruction and measurement study. AB - The postnatal development of the Eustachian tube (ET) and its surrounding structures was investigated by means of computer-aided three-dimensional (3-D) reconstruction methods in 13 normal human temporal bones, obtained from individuals 3 months to 71 years old. The cross-sectional area, width and height of the lumen in most of the cartilaginous portion of the ET were significantly smaller in children than in adults. In particular, there was a marked, age associated difference in the shape of the lumen in the cartilaginous portion of the ET. In adults, the cross-sectional area of the lumen declined monotonically between a large opening at the pharyngeal orifice and the narrowest portion of the ET (near the border of the cartilaginous and junctional regions). In children, by contrast, the ET lumen was uniformly smaller over the first 80% of its length from the pharyngeal orifice. It is suggested that this immature morphology of the ET lumen may confer increased risk of developing otitis media during childhood. PMID- 9870633 TI - Changes in nasal airway dimensions in infancy. AB - Thirty-nine infants, previously examined as neonates, were re-examined at 1 year of age with continuous wide-band noise acoustic rhinometry using a specific probe optimized for infants, to determine the dimensional growth and maturation of nasal airway geometry in otherwise healthy infants. During the first year of life, the acoustically determined dimensions of the nasal airways increased significantly. The total minimal cross-sectional area increased by 67% (0.21 cm2- >0.35 cm2), the volume of the anterior 4 cm of the nasal airway by 36% (1.80 cm3- >2.44 cm3) and the distance to the minimum cross-sectional area by 19% (0.78 cm- >0.93 cm). The rhinometric values of male infants were significantly larger than those of females. However, after adjusting for the significantly larger anthropometric values of males, the difference disappeared, indicating that it was mainly due to body size and not directly to gender. A highly significant correlation (r=0.44, p < 0.006) was observed between the minimum cross-sectional area and head circumference, which anatomically are the most closely related rhinometric and anthropometric values. Furthermore, when differentiating between infants with or without signs of nasal congestion during the fortnight preceding the rhinometric evaluation, a reduction in the total volume of the anterior 4 cm (17% p <0.02) and minimum cross-sectional area (12%, ns) was observed after adjustment. We conclude that the optimized acoustic rhinometric probe is a useful investigative modality, permitting studies of upper airway physiology of healthy and diseased infants. PMID- 9870632 TI - Evaluation of nasal patency: comparison of patient and clinician assessments with rhinomanometry. AB - The correlation between the objective measurement of nasal resistance and nasal airflow sensation is usually regarded as poor. The aim of the study was to assess the relation between objective indices of nasal patency, as assessed by the occlusion method (RN) and the Youlten peak nasal inspiratory flow meter (PNIF), with subjective sensations of nasal blockade by either the patient or the clinician in groups of patients with rhinitis, asthma, rhinitis and asthma, nasal septal deformity and ill normal controls. We studied nasal airway patency in 254 subjects (37 women, 217 men), mean age 21 years (range 14-78) by RN and PNIF. Nasal resistance was also measured by the application of Ohm's law for parallel resistors (NRO) by estimating the unilateral resistance separately. Subjective sensation of nasal blockade was assessed either by the patient on a 10-point Borg scale (SUB), or the clinician (CLN) on a 6-point scale (3 for each side of the nose). The latter was done in a controlled fashion with the aid of reference sensations. Adjusting for age, height, smoking status and airway calibre, we found good correlation between RN and CLN (r=0.57, p=10(-4)), whereas the association between RN and SUB was moderate and of borderline significance (r=0.42. p=0.05). By logistic regression analysis, we found that the only independent predictors of abnormal nasal resistance at a cut-off value of 0.30 kPa/l/s were the nasal scores as assessed by the clinician (r-=0.26, odds=2.45). We conclude that PNIF measurement and SUB scores are of limited use as indices of nasal patency, although the latter showed an improved association in comparison to older studies. As there is a necessity for some sort of objective measurement to assess nasal patency, the described clinician evaluation may be clinically useful in place of PNIF, but due to its somewhat subjective nature and its inability to detect milder levels of nasal obstruction it cannot be recommended as an alternative to rhinomanometry. PMID- 9870631 TI - Treatment of oroantral communications after tooth extraction. Is drainage into the nose necessary or not? AB - Seventy-six patients with oroantral communications after tooth extraction and chronic maxillary sinusitis were treated as follows: bacterial cultures were taken in all of them and maxillary sinuses were irrigated with an antibiotic from the cephalosporin group. Then, in 36 patients, drainage using the Caldwell Luc procedure was performed, including a naso-antral window. In all patients operations were completed by closing oroantral communications with flaps of the mucosa of the alveolar process close to the fistula. Antibiotics according to antibiogram were administered to all patients at least 10 days after surgery. Retrospective comparison between the results obtained in the first group and those in the second group 1, 3 and 6 months after operation was based on objective findings (condition of the oroantral communication, maxillary sinusitis), side effects (pain, numbness of the operated area, headache) and control radiographs (clear maxillary sinus or with mucosal thickening). The study suggests that transnasal drainage is not required in maxillary sinus surgery and in the closure of oroantral communications. Equally good results are achieved by treating with antibiotics and without drainage of the maxillary sinus into the nose. PMID- 9870634 TI - Antigen-specific IgA response of NALT and cervical lymph node cells in antigen primed rats. AB - Using antigen-primed rats, we examined the occurrence of antigen-specific antibody forming cells in mononuclear cells (MNC) of nasopharynx associated lymphoid tissues (NALT) and cervical lymph nodes (CLN) following in vitro stimulation with lipopolysaccharide (LPS), cytokines and antigen using the ELISPOT assay method. The rats were immunized with ovalubumin (OVA) by intraperitoneal injections, then by daily intranasal boostering for 2 weeks. In the nasal lavage fluid, the level of OVA-specific IgA antibody was higher and that of OVA-specific IgG antibody was lower than those antibodies in the serum. Following incubations. most of the IgA spot forming cells (SFC), but only 5 10% of IgG SFC were proved to be antigen-specific in the NALT and CLN MNCs. Following LPS stimulation, both the antigen-specific IgA and IgG SFCs increased 1.5- to 2 fold in the NALT MNC. However, following OVA stimulation, the antigen-specific IgA SFC increased 6- to 8-fold, which was remarkably higher than the antigen specific IgG SFC. The changes of the antigen-specific immunoglobulin SFCs in the CLN MNC following the antigen stimulation were similar to those in the NALT MNC. These findings suggested that NALT might be one of the induction sites for antigen-specific IgA responses in rats. PMID- 9870635 TI - Enhancement of nasal clearance of nontypeable Haemophilus influenzae by oral immunization with outer membrane proteins. AB - BALB/c mice were orally immunized with liposomes containing outer membrane proteins (OMPs) isolated from nontypeable Haemophilus influenzae (NTHi) and GM-53 as an adjuvant daily for 14 days. Anti-OMP IgA antibody titers in nasal wash, saliva, and fecal extract were significantly increased after the immunization. Although anti-OMP IgM and IgG antibodies were detected in serum, serum IgA antibodies specific to OMPs were not found. Enzyme-linked immunospot assay showed an increased number of OMP-specific IgA-secreting cells in nasal passages, intestinal lamina propria lymphocytes, and the spleen. Following oral immunization, a suspension of live NTHi was injected into the nose; nasal washes were collected 12 h after the inoculation. The number of NTHi in nasal washes was significantly reduced in mice immunized with liposomes containing OMPs and GM-53 compared to that in mice immunized with liposomes containing GM-53 alone. There was a significant negative correlation between the number of NTHi and anti-OMP IgA antibody titers in nasal washes. These findings suggest that antigen-specific IgA responses in the nose can be induced by oral immunization with OMPs and might be associated with the ability to clear NTHi from the nose. PMID- 9870636 TI - Persistent dysphagia after laser uvulopalatoplasty: a videoradiographic study of pharyngeal function. AB - In a follow-up study of 79 patients two years after laser uvulopalatoplasty 21 (27%) reported persistent postoperative dysphagia, with aspiration symptoms in 22%. None of the patients had suffered from recurrent pneumonia. A total of 4% of the patients regretted the treatment because of their dysphagia problems. The objective of this study was to examine oral and pharyngeal function videoradiographically during swallowing in the patients with persistent dysphagia, to determine whether the subjective symptoms of dysphagia correlated with objective signs of pharyngeal dysfunction. Pharyngeal function during swallowing was deviant in 76% of the dysphagic patients. In 52% of the dysphagic patients premature leakage of bolus down to different levels of the pharynx, from the tongue base to sinus piriformis, was observed before the swallowing reflex was elicited. In the dysphagic patients substantial bolus retention was observed on the epiglottis or in the valleculae alter the propagation wave had passed (43%) as well as epiglottal dysmotility (24%). Of the dysphagic patients, 10% could not avoid aspiration during the examination. These findings could explain the symptoms reported by the patients. PMID- 9870637 TI - Parvalbumin distribution in the musculature of the pharyngo-oesophageal segment. AB - The composition and size of muscle fibre types in the hyopharyngeus (HP), thyropharyngeus (TP), cricopharyngeus (CP) and cervical oesophageal muscle (CE) from 6 normal adult cats were investigated using parvalbumin (PA) immunohistochemistry. Fibre types I, IIA and IIB were identified in all muscles. HP and TP revealed predominance of type II fibres (74.8% and 75.2%, respectively), whilst CP and CE showed predominance of type I fibres (72.6% and 82.2%, respectively). The mean diameter of narrow fibres was greater in type II (23.9 microm) than in type I fibres (21.7 microm). The results seem to reflect the physiological features of each muscle, i.e. short rapid contractions of HP and TP, sustained contraction of CP and slow peristaltic movements of CE. PMID- 9870638 TI - Asymmetry of the laryngeal reflex responses to superior laryngeal nerve stimulation unrelated to the length of the recurrent nerves in the porcine model. AB - Electrical stimulation of the superior laryngeal nerve (SLN) can elicit reflex responses in the cricothyroid (CT) and thyroarytenoid (TA) muscles. We made bilateral recordings of the responses evoked in these muscles in piglets by the stimulation of either the right or the left superior laryngeal nerve (SLN). The stimulus intensity was gradually increased to study the "persistence" of the responses. We observed a direct, ipsilateral response in the CT muscle, and reflex, ipsilateral and crossed responses in both CT and TA muscles. The ipsilateral or contralateral responses obtained in TA muscles, following stimulation of the left SLN, were significantly delayed in comparison with those evoked by stimulation of the right SLN. This delay cannot be explained by the difference in length between the right and the left recurrent laryngeal nerves, but rather by an asymmetry in the sensory afferent pathway. The functional significance of this observation remains to be determined. PMID- 9870639 TI - Disease stage related in vitro responsiveness of peripheral blood T-lymphocytes in patients with head and neck carcinoma. AB - The in vitro responsiveness of peripheral blood mononuclear cells (PBMC) T lymphocytes was studied in 81 patients with limited or extended head and neck squamous cell carcinoma (HNSCC), as judged by T, N and T + N stages. Patients included in the study were males below 80 years of age, without auto-immune disease or cachexia, who were not taking any immuno-active medication at the time of diagnosis. The patients were divided into groups according to TNM stage T0-2 vs T3-4, N0-1 vs N2-3 or T + N0-3 vs T + N4-7. When cells from patients with early and late stage, according to T, N or T + N stage, were compared, we found a decreased level of mitogen stimulated T-cells and decreased spontaneous proliferation with increasing disease stage. The same was true if the in vitro mitogenesis of T-cells was analysed separately, depending on the laryngeal or oral cavity/pharyngeal origin of the patients' tumours. If the patients were divided into two groups based on N stage, decreased gamma-interferon, and to some extent interleukin (IL-2), but not IL-4 levels, were found to be related to the disease stage. PMID- 9870641 TI - Effect of unilateral section of cervical afferent nerve upon optokinetic response and vestibular nystagmus induced by sinusoidal rotation in guinea pigs. AB - An experiment was carried out in guinea pigs to clarify the effect of unilateral section of cervical afferent nerve (C1-C3) upon optokinetic nystagmus and vestibular nystagmus induced by sinusoidal rotation. To produce optokinetic nystagmus and optokinetic after-nystagmus, a random dot pattern was utilized as visual stimulation at a speed of 30 degrees/s. As for vestibular nystagmus, sinusoidal rotation at a frequency of 0.1 Hz with an amplitude of 120 degrees was used. Results showed that for about a week after the surgical section of the C1 C3 nerves, directional preponderance of the vestibular nystagmus was found toward the lesion side, whereas no significant change was obtained in optokinetic nystagmus and optokinetic after nystagmus. This asymmetric change of the vestibular nystagmus was compensated for within a week or two. Thus. unilateral section of the cervical afferent nerve produced only a temporary effect on the vestibulo-ocular reflex but it had no significant effect on the optokinetic response. PMID- 9870640 TI - Patterns of framework invasion in patients with laryngeal cancer: correlation of in vitro magnetic resonance imaging and pathological findings. AB - Total laryngectomy for laryngeal squamous cell carcinoma means a drastic change in the way of living for a patient. New surgical techniques such as laser surgery attempt to save the voice. To be oncologically correct, an accurate assessment of the tumor extent is necessary. Imaging is especially important in determining tumor extent in the regions where conventional and endoscopic ENT examinations are less accurate. Correlations of CT, in vivo MRI and pathological findings after surgery have demonstrated that MRI is more sensitive than CT, but that it overestimates the degree of cartilage invasion. Cartilage invasion is believed to be a contraindication to radiation therapy and voice-sparing surgery. In a prospective study, Gd-enhanced in vitro MRI of 10 total laryngectomy specimens was correlated with subsequent pathological examination. Good correlation of the anatomical relationships of the tumor between the in vitro images and gross pathology were found. Important is the absence of false negatives in our study, indicating that cartilage invasion can be ruled out when a normal signal intensity on in vitro MRI of the cartilage is seen. This has important oncological implications for partial voice-sparing laryngeal surgery. PMID- 9870643 TI - Significance of hearing preservation in acoustic neuroma surgery. AB - In the past 10 years, 43 patients with acoustic neuroma have been operated on by the middle cranial fossa approach. In all cases, meticulous care was taken to preserve the cochlear function regardless of the degree of preoperative hearing disorder. Thirty-nine of 43 patients had various degrees of residual hearing, hearing was preserved in 16 of these (41%). The best result was obtained in patients with a tumor located less than 1 cm from the porus. The hearing preservation rate was 64% (11/17). The patients recovered hearing acuity quite well, especially at low frequencies in 2 cases who had a pure tone hearing level of more than 50 dB and less than 50% of speech discrimination. These 2 cases had a history of sudden deafness that was intractable 5 months preoperatively. Thus it is not correct to set certain criteria for hearing preservation for patients whose preoperative hearing may not be serviceable. PMID- 9870642 TI - p53 tumor suppressor gene in acoustic neuromas. AB - Alterations of the p53 tumor suppressor gene in 21 cases with acoustic neuroma were investigated by polymerase chain reaction-restriction fragment polymorphism (PCR-RFLP) and single strand conformation polymorphism (PCR-SSCP). Neither mutation nor deletion was found. In 13 informative cases, no loss of heterozygosity (LOH) was confirmed. Thus our results further substantiate the scant contribution of p53 gene tumorigenesis and cell proliferation to acoustic neuromas. PMID- 9870644 TI - An electrophysiological study of pterygopalatine ganglion neurons in the rabbit. AB - A preparation was developed to investigate the synaptic connections of the pterygopalatine ganglion (P.P.G.) neurons of the rabbit. Many neurons received synaptic inputs from more than one preganglionic fiber in the vidian nerve (preganglionic nerve) with a wide range of conduction velocities. It is assumed that P.P.G. neurons integrate synaptic inputs from the higher centers. In some neurons. nicotinic fast excitatory postsynaptic potentials (e.p.s.p.s) were evoked in response to stimulation of one posterior nasal nerve, and also an antidromic action potential occurred in response to stimulation of the other posterior nasal nerve. Fast e.p.s.p.s were recorded from a ganglion neuron in response to the cooling stimulation of the nasal mucosa. These results revealed that the ganglionic reflex are mediated through the nasal afferent fibers exists in the P.P.G. Moreover, the appearance of slow inhibitory postsynaptic potentials (slow i.p.s.p.s) and slow excitatory postsynaptic potentials (slow e.p.s.p.s) in response to repetitive stimuli of the vidian nerve may influence the synaptic transmissions of P.P.G. neurons. The P.P.G. plays a significant role as a complicated key point of signal transmissions from both the periphery and higher centers. PMID- 9870645 TI - Effect of female hormones on the production of IL-4 and IL-13 from peripheral blood mononuclear cells. AB - In this study we compared the concentrations of IL-4, IL-13, and IFN-gamma, which were produced by human peripheral blood mononuclear cells (PBMC) in the presence or absence of preincubation with beta-estradiol or progesterone both after a specific antigen challenge and without a specific antigen challenge. The concentrations of IL-4 and IL-13 from PBMC which had been preincubated with progesterone or gamma-estradiol for 18-24 h were significantly greater than those of IL-4 and IL-13 from PBMC which had been preincubated with PBS, the control. On the other hand, the concentration of IFN-gamma from PBMC was unchanged. We were able to confirm that the female hormones beta-estradiol and progesterone, at levels similar to those occurring during pregnancy, have the ability to induce production of IL-4 and IL-13 in human mononuclear cells. These results suggest that female hormones may aggravate nasal allergy symptoms during pregnancy by increasing IgE synthesis and inducing selective eosinophil infiltration. PMID- 9870646 TI - The effect of ramatroban (BAY u 3405), a thromboxane A2 receptor antagonist, on nasal cavity volume and minimum cross-sectional area and nasal mucosal hemodynamics after nasal mucosal allergen challenge in patients with perennial allergic rhinitis. AB - The thromboxane A2 receptor antagonist, ramatroban (BAY u 3405), was orally administered for 4 weeks at a daily dose of 150 mg (b.i.d.) to 10 patients with perennial allergic rhinitis who had a positive reaction to house dust challenge on nasal mucosa. Nasal cavity volume and minimum cross-sectional area were measured, and changes in nasal mucosal swelling were determined following allergen challenge with house dust. The influence on nasal mucosal hemodynamics was also investigated. Nasal cavity volume and minimum cross-sectional area were measured by acoustic rhinometry, and blood flow in the nasal mucosa was measured by laser Doppler flowmetry. Percent changes in values from baseline nasal cavity volume were significantly decreased by allergen challenge before ramatroban administration, but no significant decrease was noted after ramatroban administration. Similarly, percent changes in values from baseline nasal cavity minimum cross-sectional area were significantly decreased by allergen challenge before administration of ramatroban, but not after administration. Percent changes in values from baseline nasal mucosal hemodynamics were significantly increased by allergen challenge both before and after ramatroban administration, which thus had no effect on mucosal hemodynamics. These findings suggest that ramatroban might inhibit the increase in nasal mucosal swelling but has no effect on nasal mucosal hemodynamics. PMID- 9870647 TI - Changes in nasal responsiveness to histamine and to specific antigen after laser surgery. AB - An initial treatment with several kinds of anti-allergic medicines is useful for reducing nasal allergy symptoms in patients suffering from Japanese cedar pollinosis during the pollen season. Since laser surgery before the pollen season seems to have a preventive effect as well, it would be of interest to know the time course of changes in the nasal reactivity to specific and non-specific stimuli after laser surgery. In this study, we investigated the changes in the nasal reactivities to specific antigen and histamine after CO2 laser surgery. The nasal reactivities to both specific antigen and histamine were enhanced 2 weeks after the laser surgery. On the other hand, they were significantly reduced after 4 weeks. Our data strongly suggest. therefore. that laser surgery must be done more than 4 weeks before the start of the pollen season to avoid temporary enhancement of nasal allergy symptoms. PMID- 9870648 TI - Influence of nasal obstruction on obstructive sleep apnea. AB - Nasal obstruction, either partial or complete, may cause snoring and/or obstructive sleep apnea (OSA). However, there is no clear answer to the question, in how many cases nasal airway is a main or combined site of obstruction. Fifty four OSA cases (47 men and 7 women) were examined by measuring the upper airway pressure gradient while they were asleep, using a multi-sensor pressure transducer. Their average apnea + hypopnea index was 35.6/h and body mass index was 26.5 kg/m2. Our hypothesis was that elevation of the inspiratory negative airway pressure at the epipharynx could indicate the influence of nasal obstruction. Seven cases out of 54 proved to have inspiratory negative pressure more than -3 cmH2O at the epipharynx during sleep. Four cases indicated elevated negative pressure in the lateral position, and 3 cases out of 19 CPAP candidates showed the same elevation in the supine position. In all 7 cases, the main obstructive site was either at the soft palate or the base of the tongue. Although nasal airway is the preferred route of respiration during sleep, nasal obstruction itself cannot be a main factor causing OSA, but a co-factor of OSA during sleep. PMID- 9870649 TI - Acoustic analysis of snoring and the site of airway obstruction in sleep related respiratory disorders. AB - Seventy-five adult patients with sleep related respiratory disorders were examined by polysomnography with simultaneous recordings of the intraluminal pressure of the upper airway and snoring sound. Obstructed sites in the upper airway during sleep were determined by comparing the amplitude of respiratory fluctuation of the pressures in the epipharynx, mesopharynx, hypopharynx and esophagus. A definite correlation existed between the intensity of snoring sound and the amplitude of respiratory fluctuation of the intraesophageal pressure. Based on the results of the intraluminal pressure partitioning, the subjects were divided into the soft palate type (28), the tonsil/tongue base type (14), the combined type (27) and the larynx type (6). The average value of fundamental frequency (ff) was 102.8+/-34.9 Hz in the soft palate type, 331.7+/-144.8 Hz in the tonsil tongue base type, 115.7+/-58.9 Hz in the combined type and around 250 Hz in the larynx type. PMID- 9870650 TI - Evaluation of uvulopalatopharyngoplasty in treatment of obstructive sleep apnea syndrome. AB - Many methods have been reported to select the proper candidates for uvulopalatopharyngoplasty (UPPP) among obstructive sleep apnea patients. Polysomnography with intraluminal pressure recordings in the upper airway was found useful to predict obstructive site(s) and hence the success of surgery. According to the pressure analysis, 11 cases with high negative pressure at the mesopharynx were selected for UPPP, while UPPP combined with midline partial glossectomy (UPPP-MLG) was done in 18 cases proved to have high negative pressure difference between the mesopharynx and esophagus. A reduction of the apnea hypopnea index (AHI) of more than 50% after UPPP was found in 8 cases (72.7%) and in 11 cases (61.l%) after UPPP-MLG. It was found that the response rate was lower among the second group if compared to that of the first because of other aggravating factors rather than simple obstruction at the level of the soft palate and or tonsils-tongue base level. PMID- 9870651 TI - Analysis of long-term results of our combination therapy for squamous cell cancer of the maxillary sinus. AB - Seventy-four patients between 1971 and 1982 in Period I and 32 patients between 1982 and 1987 in Period II with maxillary sinus squamous cell cancer were treated by combination therapy consisting of preoperative LINAC X-ray irradiation with 5 flourouracil intra-arterial chemotherapy followed by maxillectomy and primary reconstruction. In Period II, 21 patients received preoperative cisplatinum (CPPP) microcapsule chemoembolization and pepleomycin (PEP) i.m. and or postoperative CDDP i.v. with PEP i.m. in addition to the combination therapy administered in Period I depending on systemic conditions, tumor stage and histopathological type. Five and 10-year crude survival rates were 68.9% and 55.4%, respectively, for Period I and 71.9% and 56.3% for Period II, with no significant difference between the two trials. In 21 selected patients in Period II, who had additional chemotherapy with preoperative CDDP chemoembolization and/or postoperative i.v. infusion of CDDP with pepleomycin i.m., 5 and 10-year survival rates were 85.7% and 61.9%, respectively. PMID- 9870652 TI - Analysis of factors affecting long-term treatment results of adenoid cystic carcinoma of the nose and paranasal sinuses. AB - At Akita University Hospital (from 1971 to 1987) and at Chiba University Hospital (from 1983 to 1988), 17 patients with adenoid cystic carcinoma of the nose and paranasal sinuses were treated by en bloc tumor resection followed by primary reconstruction of the maxilla. Pre- and postoperative radiation was combined in 5 and 6 patients, respectively. Ten-year cancer-free survival rates were 59.3% in the 12 patients with maxillary sinus tumors and 50% in the 4 patients with nasal tumors. One patient with a sphenoid sinus tumor died within 5 years. Ten-year cancer-free survival was affected markedly by grade of tumor extension. Among T1N0M0 and T2N0M0 patients (of which there were 1 and 7, respectively), only 1 died of unrelated causes, and 6 patients survived cancer-free for more than 10 years. However, 4 of the 6 T3N0M0 patients died, and the cause of death was distant metastasis in 2, intracranial tumor extension in 1, and unrelated causes in 1. All 3 T4N0M0 patients died, 2 due to intracranial tumor extension and 1 of unrelated causes. The cause of death was distant metastasis in 3, local recurrence in 3, 2 of whom had intracranial tumor extension, and unrelated causes in 2. Prevention of distant metastasis and intracranial tumor extension was considered to be crucial for improving treatment results after en bloc tumor resection. Preoperative radiation was thought to decrease incidence of distant metastasis. In 5 patients who had preoperative radiation. 4 survived cancer-free for more than 10 years, and only 1 patient died of unrelated cause. However. of the 6 patients who had postoperative radiation, 2 died of distant metastasis and another 2 died of intracranial tumor extension. Of the 6 patients who did not have radiation therapy, the causes of death were local recurrence in 1, distant metastasis in 1 and unrelated causes in 1. Preoperative radiation in 5 patients showed histopathologically moderate or marked degeneration and necrosis of tumors in all patients. Although the number of patients in this study is too small to allow statistical analysis of the data, our present modality of treatment for adenoid cystic carcinoma of the nose and paranasal sinuses is routine combination of preoperative full dose radiation, en bloc tumor resection and primary reconstruction, including en bloc resection of the cranial base in selected T4 patients. PMID- 9870653 TI - Surgical techniques for benign parotid tumors: segmental resection vs extracapsular lumpectomy. AB - In 80 patients surgical techniques for benign parotid gland tumor have been reviewed and evaluated. Segmental resection was introduced during partial superficial parotidectomy. The superficial lobe of the parotid gland was divided into four sections according to the distribution of the tumors. Total parotidectomy was applied to remove deep lobe and recurrent tumors, together with the superficial lobe, followed by retrograde dissection of the facial nerve. This technique was termed the hamburger technique. Furthermore, with a margin of normal tissue, extracapsular lumpectomy was used. Segmental resection is at method of extirpation by dissecting the facial nerve, whereas extracapsular lumpectomy is a method of extirpation by dissecting the tumor itself with a safety margin. PMID- 9870654 TI - Metastasis of renal cell cancer to the ethmoid sinus. AB - The origin of metastatic tumors in the nasal or paranasal sinuses is often renal cancer, and metastasis to the nasal or paranasal sinuses sometimes takes a long time after nephrectomy. The present paper deals with one patient with metastasis of renal cancer to the ethmoid sinus 2 years after nephrectomy. Even though many metastatic tumors originating from renal cancer develop in multiples, most metastatic tumors in the nasal or paranasal sinuses are single and treated surgically. However, even if multiple tumors are found in the nasal and paranasal region and other organs, surgery will be effective in preventing epistaxis and subsequent anemia. Also, when removing a tumor it will be effective to identify the feeding arteries of the tumor, perform embolization therapy, and clip the necessary arteries. PMID- 9870655 TI - Studies on binding of HIV-1 p24gag peptide to HLA-Cw3+ cells. AB - Human major histocompatibility complex class I antigens, HLA-C, are expressed on the cell surface at approximately a tenfold lower level than HLA-A and -B. We hypothesized that the expression of HLA-C is limited by the quantity of high affinity peptides which bind to these molecules, thus allowing only a small fraction of HLA-C molecules to be transported and/or to remain stable on the cell surface. If this assumption is correct, then the addition of exogenous peptide should increase cell surface HLA-C expression. To verify the hypothesis, we pulsed lymphoblastoid cell line PAJ (HLA-Cw3+) with synthetic HIV-1 p24gag 145 152 peptide, known to be presented to T-lymphocytes by HLA-Cw3 molecule. PAJ (HLA Cw3+) cells bound approximately two times more of the peptide than HAJ (HLA-Cw3 ), and four times more than 500/C9 (HLA-Cw3-) cells. Accordingly, overnight pulsing of PAJ cells with the p24gag 145-152 peptide caused an increase in class I HLA expression detected on the cell surface by flow cytofluorimetric analysis with anti-HLA-B,C monoclonal antibodies but not by anti-HLA-A antibody. In contrast, HLA-Cw3- cells treated in the same manner did not show any increase of HLA class I expression. Our data suggest that low concentration of high affinity peptides within the cell may be one of the factors limiting cell surface expression of HLA-C molecules. PMID- 9870657 TI - Lysis of CD4+ T cells expressing HIV-1 gag peptides by gag-specific CD8+ cytotoxic T cells. AB - The vast majority of in vitro experiments testing the cytotoxic T lymphocytes (CTL) activity in HIV infection has been performed with target cells consisting of autologous EBV-transformed B lymphoblastoid cell lines (B-LCLs) expressing Human immunodeficiency virus type I (HIV-1) proteins. However data concerning the lysis of primary CD4+ T lymphocytes expressing HIV-1 antigens by CTLs is still lacking. To study the CTL activity against such primary targets, we used a system involving PBMCs of an HIV+ asymptomatic patient (PT) as effector cells and the CD4+ lymphocytes or B-LCLs of his healthy HLA-identical twin brother (HTW) as target cells. These syngeneic targets were either infected with recombinant vaccinia virus containing HIV-1 gag gene (gag-vac), or coated with HIV-1 gag peptides. We demonstrate in this study that PT CTLs (which were CD3+, CD4-, CD8+, TCRalphabeta+, TCRgammadelta-, CD56-) specifically lysed both types of syngeneic target cells expressing gag-vac; however, CD4+ T cells expressing HIV gag proteins were lysed less efficiently than B-LCLs expressing the same HIV epitopes. On the other hand, no specific lysis was detected when the target cells were uninfected or infected by wild-type vaccinia virus. PMID- 9870656 TI - Cytomegalovirus-specific IFNgamma and IL-4 are produced by antigen expanded human blood lymphocytes from seropositive volunteers. AB - The cytokine responses to cytomegalovirus (CMV) antigen in seropositive and seronegative individuals were measured using a combination of antigenic expansion and intracellular staining. Intracellular IFNgamma and IL-4 were produced in a dose-dependent manner by T cells in response to CMV only in the seropositive population. The potential for individual cells to produce both Th1 and Th2 cytokines simultaneously was clear, as IL-4 was most often produced in those cells with the highest IFNgamma production. The cytokine-specific nature of this response was demonstrated by fluorescence microscopy, which showed granular cytoplasmic staining, and at the mRNA level by ribonuclease protection assays. These methods expand our ability to evaluate the immune response to CMV, and can now be correlated to a number of clinical conditions. PMID- 9870658 TI - MPK38 expression is upregulated in immature T cells activated by concanavalin A. AB - We have previously identified a cDNA clone from a murine teratocarcinoma PCC4 cDNA library, which we have termed MPK38 for murine protein kinase 38. MPK38 is a new member of the SNF1 serine/threonine kinase family. To understand the role of the MPK38 in thymocytes, we have investigated the level of MPK38 expression by Northern blot analysis. Interestingly, incubation of thymocytes at 37 degrees C resulted in the loss of the MPK38 transcript, however the transcript could be reinduced by treatment with Con A or PHA, but not with PMA and growth factors such as IL-2, IL-7, TGF-beta, TNF-alpha, and EGF. In addition, stimulation of mature T cells from the spleen with Con A failed to reinduce the MPK38 transcript, indicating a developmental regulation of MPK38 expression, whereas non-T cell populations significantly reinduced the transcript. These results suggest that MPK38 may play a functional role as one of the signal regulators in early T cell activation and, potentially, certain lineages of hematopoietic cell activation induced by Con A. PMID- 9870659 TI - TCR diversity in gammadeltaTCR+ hybridomas derived from mice given portal vein donor-specific pre-immunization and skin allografts. AB - Portal venous (p.v.) immunization with multiple minor histoincompatible cells leads to antigen-specific increased skin allograft survival. GammadeltaTCR+ hybridoma cells, prepared from mesenteric lymphocytes of p.v. immunized animals, can adoptively transfer this increased graft survival to naive animals. We have analyzed VgammaVdelta gene usage, and TCR gamma-chain junctional diversity in gammadeltaTCR+ hybridomas from mice immunized with different antigen combinations by p.v. or conventional lateral tail vein (i.v.) immunization. Following p.v. immunization two independent sets of hybridoma cells were derived, one expressing a common gamma-chain junctional sequence which was also found in > 85% of the hybridomas derived following i.v. immunization, while the other set showed remarkable gamma-chain junctional sequence diversity. The diversity seen in these latter hybridomas was associated with the antigen specificity of the hybridoma cells. Cells expressing these 'unique' TCR junctional sequences were stimulated to produce cytokines both by hsp and by minor-histocompatibility-specific irradiated peritoneal cells. Cells expressing TCR with a common gamma-chain junctional sequence were stimulated to cytokine production by MHC-matched but minor-histocompatibility mismatched (as well as matched) peritoneal cells, but not by hsp. We suggest that p.v. immunization results in stimulation of both antigen-specific and non-specific regulatory gammadeltaTCR+ cells, which can be distinguished by gamma-chain TCR sequence diversity. PMID- 9870660 TI - Role of transmembrane domains in the functions of B- and T-cell receptors. AB - The antigen receptors on the surface of B- and T-lymphocytes are complexes of several integral membrane proteins, essential for their proper expression and function. Recent studies demonstrated that transmembrane (TM) domains of the components of these receptors play a critical role in their association and function. It was specifically demonstrated that in many cases point mutations in the TM domains can partially or completely disrupt the receptor surface expression and function. Here we review studies of the TM domains of B- and T cell receptors. Furthermore, we use a novel method, PHDtopology, to provide estimates of the exact locations and lengths of the TM domains of the subunit components of these receptors. Most previous studies used single residue hydrophobicity as a criterion for determining the position and length of the TM domains. In contrast, PHDtopology utilizes a system of neural networks and the evolutionary information contained in multiple alignments of related sequences to predict the location, length, and orientation of transmembrane helices. Present results significantly differ from most published estimates of the TM domains of the B- and T-cell receptor components, primarily in the length of the TM domains. These results may lead to modification of putative TM motifs and re interpretation of the results of studies using mutated TM domains. The availability of PHDtopology on the Internet would make it a valuable tool in the future studies of the TM domains of integral membrane proteins. PMID- 9870661 TI - Defective degranulation and calcium mobilization of bone-marrow derived mast cells from Xid and Btk-deficient mice. AB - Regulation of adhesion and degranulation of mast cells plays an important role in allergy and inflammation. We investigated a possible role of Bruton's tyrosine kinase (Btk) in the regulation of adhesion and degranulation by using bone marrow derived mast cells from X-linked immunodeficiency (Xid) and Btk-deficient mice. Cross-linking of the high affinity IgE receptor (Fc epsilonRI) and steel factor (SLF) induced indistinguishable adhesive responses of mast cells to fibronectin in kinetics, and these adhesive responses were comparable among wild type, Xid, and Btk-deficient mast cells. Cross-linking of Fc epsilonRI, but not SLF triggered degranulation of bone marrow-derived mast cells. However, Fc epsilonRI induced degranulation was impaired in Xid and Btk-deficient mast cells. Calcium influx induced by Fc epsilonRI cross-linking and SLF were also reduced in Xid and Btk-deficient mast cells. Degranulation and calcium influx were reduced more severely in Btk-deficient than in Xid mast cells. Consistently, cross-linking Fc epsilonRI and SLF augmented Btk kinase activities transiently. Inositol triphosphate (IP3) production was also severely reduced in Btk-deficient mast cells, indicating Btk play a critical role of Fc epsilonRI-induced IP3 production. The differential sensitivity of wortmannin on calcium influx in wild type and Xid mast cells suggested that the activation of phosphatidylinositol 3 kinase (PI 3-kinase) was required in calcium influx. Furthermore, abnormal secretory granules with translucent contents and variable in size were observed both in Xid and Btk-deficient mast cells. Our study demonstrated a critical role of Btk in regulating intracellular calcium and granule exocytosis. PMID- 9870662 TI - Ragweed allergy: correlation between skin reactivity and in vitro complement activation. AB - Recently we have reported on several observations which indicate that allergen induced complement activation contributes to the development of the symptoms of ragweed allergy. In the present paper a new finding that supports this assumption is summarized. In 48 ragweed-allergic patients individual skin reactivity to ragweed allergen extract (RWA) was assessed using dilution skin prick testing. Sera of these patients were incubated with 20, 100, and 400 U/ml RWA and generation of two complement activation products, alternative pathway C3 convertase (C3bBbP) and terminal pathway activation complex (C5b-9) was measured by ELISA methods. A strong positive correlation (Spearman correlation coefficient r = 0.495, P = 0.0004, and r = 0.454, P = 0.0012, respectively) was found between individual skin reactivity to RWA and C3bBbP generation induced by 20 and 100 A allergological units/ml (U/ml) RWA. This finding further supports the role of complement activation products in the aggravation of the basic IgE-mediated immunopathology of ragweed allergy. PMID- 9870663 TI - The effects of interleukin-15 on human gammadelta T cell responses to Plasmodium falciparum in vitro. AB - We observed that the gammadelta T cell subset expands when human peripheral blood mononuclear cells (PBMC) from malaria-naive donors are cultured with Plasmodium falciparum lysate in the presence of IL-2 or IL-15, cytokines that utilize two common IL-2 receptor subunits. IL-15 induced the expansion of the gammadelta T cell subset at all levels tested, whereas IL-2 was not stimulatory at high levels. Flow cytometric analysis of apoptosis using the TUNEL assay indicated that the percentage and absolute number of gammadelta T cells undergoing apoptosis were greater in cultures stimulated with antigen and IL-2 than in cultures stimulated with either antigen and IL-15 or control erythrocyte lysate and IL-2. The ability of IL-15 to enhance gammadelta T cell function was also assessed; the results suggest that IL-15 can function with IL-2 to enhance the capacity of gammadelta T cells to inhibit parasite replication. Together these data indicate that IL-2 and IL-15, which both bind to IL-2Rbeta and IL 2R(gamma)c, enhance gammadelta T cell function, but they appear to have different effects on proliferation and survival. PMID- 9870664 TI - Beta-selection of immature thymocytes is less dependent on CD45 tyrosinephosphatase. AB - Tyrosine kinase p56lck plays a pivotal role in beta-selection from CD4-8- (DN) to CD4+8+ (DP) developing pathway, but it is unclear how CD45 transmembrane tyrosinephosphatase is involved in this process although CD45 activates p56lck by dephosphorylating its tyrosine-505. To analyze this issue, we produced double mutant mice of T-cell receptor transgenic mice (TCR-Tg) or RAG-2 knock out mice backcrossed with either p56lck or CD45 knock out mice. In TCR-Tg, CD25+DN thymocytes almost disappeared and CD25-44-DN cells of further developing stage increased, implying that all DN thymocytes can undergo beta-selection due to the expression of functionally rearranged TCR-beta on CD25+ DN thymocytes. However, CD25+ thymocytes increased in DN stage when TCR-Tg were backcrossed with p56lck deficient mice but not with CD45 deficient mice. Similarly, DP thymocyte induction with CD25+ cell reduction in RAG-2 knock out mice by injection of anti CD3 mAb was inhibited in p56lck deficient but not in CD45 deficient mice. This suggests that CD45 is dispensable for beta-selection though p56lck is required. PMID- 9870665 TI - Construction and expression of a mouse-human chimeric antibody against human tumor necrosis factor-alpha. AB - A mouse anti-human tumor necrosis factor-alpha (TNF-alpha) monoclonal antibody (MoAb), designated as 3B10, has previously been produced and characterized by our laboratory. We report here the construction and the expression of mouse-human chimeric antibody derived from the MoAb. cDNAs encoding variable regions of heavy and light chains were prepared from 3B10 cells by polymerase chain reaction, and introduced to mammalian expression vectors containing cDNA for human gamma1 and kappa constant regions, respectively. Cotransfection of the vectors into CHO cells resulted in production of antibody reacting with human TNF-alpha. In SDS PAGE analysis, the chimeric antibody, c3B10, migrated at 170 kDa under a nonreducing condition, whereas two bands with 58 and 28 kDa appeared following treatment with 2-mercaptoethanol. Both c3B10 and mouse 3B10 neutralized the cytotoxic activity of human TNF-alpha to the same level, indicating that c3B10 holds the binding activity of its original MoAb. These findings suggest that the introduced genes for chimeric heavy and light chains are transcribed and translated to produce the chimeric heavy and light chain peptides, and that the peptides are assembled to form native IgG molecule. The chimeric anti-TNF-alpha antibody described in this study is expected to be less immunogenic and thus more suitable for possible clinical use. PMID- 9870666 TI - The role of polymorphonuclear leukocytes in the resistance to cutaneous Leishmaniasis. AB - The massive infiltration by polymorphonuclear leukocytes (PMN) soon after skin infection with Leishmania major suggests that PMN could participate in reducing parasite load and controlling the spreading of leishmanial infection. Yet, direct evidence for the participation of PMN in host defense against L. major was lacking. We investigated L. major infection in susceptible and resistant mice treated with the monoclonal (mAb) antibody RB6-8C5 that depletes the population of mature neutrophils and eosinophils. Both BALB/c and C57BL/6 mice depleted of PMN show accelerated parasite spreading and more severe footpad swelling than similarly infected untreated mice. In addition, significant higher parasite numbers were found in the lesion draining lymph nodes from PMN-depleted C57BL/6 mice. Histopathological analysis of the paw confirmed neutrophils containing ingested parasites as the dominant cell type in the infiltrate of the first days after infection and the nearly absolute neutrophil depletion in mAb-treated mice. Our data show the importance of PMN in early control of parasite load and parasitism spreading in cutaneous leishmaniasis. PMID- 9870667 TI - Tumor surveillance: expression of the transporter associated with antigen processing (TAP-1) in ex-vivo human tumor samples and its elevation by in vitro treatment with IFN-gamma and TNF-alpha. AB - We have analyzed 30 human tumor specimens (two breast, six lung, and 21 ovarian carcinomas and one malignant melanoma) for the expression of the transporter associated with antigen processing, TAP-1. Cell suspensions judged to contain negligible contamination with non-tumor cells were tested for reactivity with the antibody directed to TAP-1 in Western blot. According to these results all lysates prepared from the tumor samples contained the protein, but with considerable quantitative variations. Tumors exposed in vitro for a short time to IFN-gamma and TNF-alpha had elevated TAP-1 levels in 12 out of 30 experiments. In accordance with previous results, the tumor cell populations were heterogeneous with regard to MHC class I expression, as analyzed by indirect immunofluorescence on viable cell suspensions. Short term in vitro treatment with IFN-gamma and TNF alpha elevated MHC class I expression in several tumors. In several mixed cultures, cytokine treated tumor cells induced cytotoxic activity in autologous or allogeneic lymphocyte populations. In vitro treatment with IFN-gamma and TNF alpha upregulated thus the expression of MHC class I and TAP-1 in a fraction of the tumor samples. However the potentiation of the capacity to interact with T cells induced by the cytokines was not always parallel with the changes in these two parameters. It is therefore likely that the cytokine treatment induced additional changes in the tumors which contributed to their capacity to elicit the T cell response. PMID- 9870669 TI - Tumor necrosis factor alpha (TNF-alpha) modulates rat mast cell reactivity. AB - Nowadays there is growing evidence that some cytokines regulate biological functions of the mature mast cells. Therefore, we have studied whether TNF-alpha, the cytokine of multifunctional activities, could directly stimulate rat peritoneal mast cells to histamine secretion and whether it could modulate rat mast cell reactivity in anaphylactic (with ConA) and anaphylactoid (with compound 48/80) reactions. We have established that rat recombinant TNF-alpha does not activate rat mast cells to histamine release. However, TNF-alpha-treatment causes the decrease of spontaneous histamine release up to 85% (TNF-alpha concentration: 2 x 10(-9) M). Pretreatment of mast cells with TNF-alpha inhibits ConA-stimulate release of histamine with the percent release decreasing up to 33.7% of the control value (TNF-alpha concentration: 5 x 10(-9) M) and this decrease is statistically significant. Pretreatment of mast cells with TNF-alpha reduces compound 48/80-dependent histamine release as well and the percent release of histamine fell to 64.7% of the control value. We have concluded that TNF-alpha may play a significant role in regulation of mast cell secretory activity. PMID- 9870668 TI - Effect of Bacillus firmus on antibody formation after mucosal and parenteral immunization in mice. AB - Immunostimulatory properties of B. firmus, a nontoxic, nonpathogenic G + bacterium of external environment, were described previously. Antiinfectious and antitumor activity, macrophage activation and strong polyclonal stimulation of B lymphocytes were proved in human, mice and rats. The adjuvant effect of B. firmus on specific antibody response to ovalbumin in BALB/c mice is the topic of the present study. Against our expectation, B. firmus exerts more suppressive than stimulatory effect on specific antibody response. Formolized B. firmus decreased anti-ovalbumin response after subcutaneous immunization and only slightly increased serum antibodies after intraperitoneal immunization. After mucosal immunization, both oral and rectal, ovalbumin itself did not cause a significant systemic response but induced IgA anti-ovalbumin response in the intestine. B. firmus applied together with ovalbumin increased systemic serum response but absolutely eliminated intestinal response. The rectal route of antigen administration has been found less convenient because of less precise dosing of antigen in this mode of immunization. PMID- 9870670 TI - The S100 family of multipurpose calcium-binding proteins. AB - Antibodies reactive with S100 protein are useful markers in a diagnostic immunohistochemistry laboratory dealing with cutaneous tumors. However, S100 protein is not a single protein but instead a group of S100 proteins with diverse functions. S100 proteins constitute a family of acidic calcium-binding proteins that are important in intracellular calcium metabolism. Recent evidence that some S100 proteins are secreted makes it likely that they are also involved in cell cell interactions. The exploration of the status of the different members of the S100 family may yield not only diagnostic clues but also relevant functional information about the cells. Considerable recent progress has been made in our understanding of S100 proteins. This review surveys some of these findings that may be either directly or indirectly relevant to cutaneous pathology. PMID- 9870671 TI - Multibacillary leprosy: lesions with macrophages positive for S100 protein and dendritic cells positive for Factor 13a. AB - In the defense against Mycobacterium leprae, macrophages play an essential part in the mechanism of bacterial lysis but require the presence of cytokines such as interleukin 2 and gamma interferon from lymphocytes in order to effectively kill the organisms in any number. While there have been many studies of the lymphocytes in lesions of leprosy, less attention has been given to the immunohistochemical characterization of the macrophage populations. In this study, the cutaneous lesions of 69 patients with leprosy (42 lepromatous, 5 mid borderline, and 22 tuberculoid) were evaluated by immunohistochemistry for the expression of S100 protein, CD1a, CD68, muramidase, HLA-DR, and Factor 13a. The macrophages from lesions of polar, subpolar, and borderline lepromatous leprosy patients expressed S100 protein intensely and constantly. In contrast, the lesions of polar and subpolar tuberculoid leprosy had very few cells that were immunoreactive for S100 protein ('S100+') in the granulomas in the dermis. The macrophages in all lesions were reactive for CD68 and muramidase. In paraffin sections, macrophages of lepromatous lesions failed to stain for HLA-DR, whereas in tuberculoid lesions, they were strongly positive for HLA-DR. Three patients with histoid leprosy (relapse lesions) had lesions that were strongly positive for Factor 13a and were negative for S100 protein ('S100-'). Given the possible chemotactic and migration inhibition effects of the calcium-binding proteins of the S100 family, these data suggest a possibly important role for S100 protein in the accumulation of macrophages in lepromatous leprosy, and also reveal infection of Factor 13a + dermal dendritic cells in histoid leprosy. PMID- 9870672 TI - A distinctive cutaneous reaction pattern indicative of infection by reactive arthropathy-associated microbial pathogens: the superantigen ID reaction. AB - The two major cutaneous expressions of infective states are infections of the skin by viable organisms and immunological responses to nonviable microbial antigens or, in the case of molecular mimickry, their human analogues. These immunological responses are designated as cutaneous id reactions, and manifest a histomorphology similar to that seen at the primary infective site. This study presents the clinical and histological findings in 16 patients who developed skin eruptions associated with extracutaneous or systemic infections. There was a striking female predominance; patients ranged in age from 10 to 78 years. The majority of cases manifested skin lesions which clinically resembled Sweet's syndrome, erythema multiforme and/or erythema nodosum. Fever, arthralgia, oligoarthritis, mucosal ulcers of the mouth and/ or genital tract and uveitis were additional features in some cases. Isolated clinical presentations included a petechial rash in a stocking and glove distribution, papular dermatitis, a morbilliform eruption and annular erythema. Among the medical and family histories were atopy and stigmata associated with connective tissue disease (CTD). Two patients were ingesting drugs with known immune dysregulating properties. Skin biopsies showed focal lymphocytic interface dermatitis, a diffuse interstitial histiocytic infiltrate, and a mononuclear cell predominant vascular reaction which in some cases represented vasculitis by virtue of manifesting concomitant luminal or mural fibrin deposition. Eosinophils, eczematous alterations, and papillary dermal edema were identified in a minority of cases. All patients had evidence of a prior or concurrent infection, based on either positive IgM serology for specific microbes or cultures. Among the implicated pathogens were cytomegalovirus, parvovirus B19, streptococcus, mycoplasma, klebsiella, and Borrelia burgdorferi. All of these organisms are among those associated with reactive arthritis, a phenomenon that was seen in some cases. The histology suggested florid cell mediated immunity (CMI), which the authors attributed to the superantigen properties held by the aforesaid pathogens. Skin lesions and constitutional symptoms resolved quickly with antimicrobial therapy in 7 of 9 cases causally linked to bacteria. Spontaneous resolution occurred in 5 of 6 virally mediated eruptions. The other 4 patients were given topical steroids or prednisone; these included 1 patient with Borrelia burgdorferi infection and 1 patient with radiographic evidence of pneumonia who was never cultured, 1 patient with parvovirus B19 infection, and 1 patient with pneumococcal pneumonia and concomitant sarcoidosis. It is the authors' belief that the eruptions seen in these patients may in part reflect a genetic or iatrogenic predisposition to respond excessively to certain infectious triggers. PMID- 9870673 TI - Grover's disease (transient acantholytic dermatosis): relationship of acantholysis to acrosyringia. AB - Transient acantholytic dermatosis is often associated with excessive sweating, fever, and bed confinement. The pathogenesis of this disease has been postulated to be poral occlusion of damaged eccrine intraepidermal ducts. Histological and immunohistochemical and ultrastructural studies were performed on 10 biopsies from 10 patients with transient acantholytic dermatosis. Immunoreactions for carcinoembryonic antigen and cytokeratin-7 to identify eccrine duct epithelium were performed on all 11 biopsies. In addition, 5 of the biopsies were immunoreacted for cytokeratin 8. All immunoreactions were reviewed independently by two observers to determine extent of reactivity and whether it correlated with areas of epidermal acantholysis. Among the 11 biopsies, 8 showed acantholysis not associated with eccrine duct outflow tracts. In 2 biopsies the acantholysis was consistently associated with acrosyringea; in one case acantholysis was inconsistently associated with eccrine outflow tracts. Epidermal acantholysis in patients with Grover's disease is associated with the outflow tracts of eccrine ducts in a subgroup of patients. Although leakage of sweat from occluded sweat ducts in acrosyringia may be the mechanism operating in a subgroup of patients with Grover's disease, this does not appear to be the subgroup of patients in whom Grover's disease develops in the setting of being bedridden and/or sweating. PMID- 9870674 TI - Demodex mites in acne rosacea. AB - The hair follicle mites Demodex folliculorum and Demodex brevis and their role in the pathogenesis of rosacea have been the subject of much debate in the past. We studied the prevalence of Demodex mites in facial skin biopsies obtained from 80 patients with rosacea, 40 with facial eczematous eruption and 40 with lupus erythematosus discoides. The mite prevalence in the rosacea group (51%) was significantly higher than in the rest of the study population (eczema 28% and lupus discoides 31%). Demodex mites were found on all facial sites. The most infested areas in the whole study group were the forehead (49%) and the cheeks (44%). Males were more frequently infested (59%) than females (30%). We did not find any significant difference in mite counts of infested follicles between rosacea and the control group. A lympho-histiocytic cell infiltration was seen around the infested hair follicles. Our results suggest that Demodex mites may play a role in the inflammatory reaction in acne rosacea. PMID- 9870675 TI - Factor XIIIa-positive dermal dendritic cells and HLA-DR expression in radial versus vertical growth-phase melanomas. AB - Fifty nevomelanocytic lesions, including 10 typical compound nevi and 20 radial and 20 vertical growth-phase melanomas, were evaluated for factor XIIIa and HLA DR (LN3) expression within dermal dendritic cells (DDCs) or dermal dendrocytes to determine if DDCs proliferate and/or participate as possible antigen-presenting cells in the local tissue response to benign and malignant nevomelanocytic lesions. There was no statistical difference in factor XIIIa staining of DDCs between nevi and radial or vertical growth-phase melanomas, suggesting that DDCs do not significantly proliferate in nevomelanocytic lesions. However, studies to determine proliferation rate, apoptosis, and influences of local mediators on cell growth and/or recruitment were not done. HLA-DR staining by DDCs was significantly increased (p<0.001) in both melanoma groups when compared to compound nevi, but did not significantly differ between radial and vertical growth-phase melanomas. The intensity of HLA-DR expression appeared to correlate with the presence or absence of lymphocytic inflammation; HLA-DR intensity was judged greater in melanomas characterized by a brisk and infiltrative lymphocytic host response. We propose that DDCs may participate in the dermal immune response to invasive melanomas, probably as antigen-presenting cells to skin-associated lymphocytes. PMID- 9870676 TI - Histopathology of cutaneous reaction to granulocyte colony-stimulating factor: another pseudomalignancy. AB - Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth factor (HGF) with many applications in cancer therapy. The most important applications are reduction in the incidence of febrile neutropenia, acceleration of neutrophil recovery after chemotherapy or bone marrow transplantation, and mobilization of progenitor cells. Many cutaneous adverse reactions associated with HGF have been reported in recent years, including injection site reactions, pyoderma gangrenosum, Sweet's syndrome, cutaneous leucocytoclastic vasculitis, and widespread folliculitis. The presence of large histiocytes on the dermis between collagen bundles has been proposed as a characteristic histopathologic finding in cutaneous eruptions secondary to granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. We report on a patient with a high-risk ductal infiltrating carcinoma of the breast who received high-dose chemotherapy (HDC) with peripheral blood progenitor cell (PBPC) rescue. The patient received G-CSF after PBPC for a faster granulocyte recovery. She developed a cutaneous eruption located on back, buttocks, axillae, groin and sites where electrocardiography electrodes had been placed. From the histopathological point of view, the eruption was characterized by the presence of numerous large, atypical histiocytes in the dermis with several mitotic figures, mimicking involvement of the dermis by a malignant process. PMID- 9870677 TI - Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease): case report with nodal and diffuse muco-cutaneous involvement. AB - Sinus histiocytosis with massive lymphadenopathy (SHML) (Rosai-Dorfman disease) is a rare self-limited histiocytic disorder of unknown origin, usually present with cervical lymphadenopathy, fever, elevated ESR and haematologic abnormalities. Extranodal lesions may occur as initial manifestations of the disease. In most cases skin lesions are associated with nodal or extranodal localization. A 54-year-old male is reported who, for a period of ten months, presented numerous slowly enlarging, rounded papular lesions on the face, neck, trunk as well as severe cervical lymphadenopathy without general and haematological disturbances. Thereafter a fungating mass developed in the nasal cavity. Histological, histochemical and immunohistochemical tests of skin, mucosal and lymph-node lesions were consistent with a diagnosis of SHML. The immunohistochemical characterization of the cytophagocytic S-100+ macrophages revealed that they were highly positive for the adhesion molecules Vitronectin receptor av/b3, which is used by tissue macrophages for removal of effete cells through phagocytosis. PMID- 9870678 TI - Genital Paget's disease with clear cells in the epidermis of the axilla. AB - We report an 84-year-old man with extramammary Paget's disease (EMPD) involving the genital region. Microscopic examination revealed very few clear cells appearing pagetoid in the lower portion of prickle cell layer of the right axilla epidermis, with no clinically detectable eruption. Their histochemical, immunohistochemical and lectin-binding reactions were almost identical to those in the genital lesion. However, although the axillary lesion was diagnosed as subclinical Paget's condition, the clinical course showed no aggressive or destructive nature. Our case suggests that not all subclinical Paget's conditions become malignant, and that in some cases the clear cells may be precursors of Paget's cells developing multifocally. PMID- 9870679 TI - Pagetoid Merkel cell carcinoma: epidermal origin of the tumor. AB - We report a case of intraepidermal Merkel cell carcinoma which occurred on the face of a 76-year-old white male. This slow-growing tumor was mostly confined in the epidermis and pilosebaceous apparatus where tumor cells spread in a pagetoid fashion forming tumor cell nests. Histologically it resembled a superficial spreading melanoma. A heavy lymphocytic infiltration was seen beneath the epidermal lesion as is often seen in pagetoid melanomas. Histochemical and ultrastructural features such as the presence of cytokeratin 20, synaptophysin, neuron specific enolase, desmosomes, and dense cored granules confirmed the diagnosis of Merkel cell carcinoma. Occasional mitotic cells and many apoptotic cells were found in the tumor. Dylon positive, amyloid depositions were seen in the lower epidermis and papillary dermis; they were probably derived from apoptotic tumor cells. It was thought that apoptosis limited the speed of growth of this tumor. We believe that this is probably the most convincing case of intraepidermal Merkel cell carcinoma originating from epidermal Merkel cells or its precursors (stem cells). PMID- 9870680 TI - Acrokeratoelastoidosis of Costa: a primary disease of the elastic tissue? AB - A 24-year-old black woman from Uganda was seen for treatment of multiple papules on her hands and feet. The lesions corresponded microscopically to foci of hyperkeratosis and acanthosis. Acid orcein stain revealed marked elastic fiber fragmentation. Acrokeratoelastoidosis of Costa (AKE) was diagnosed. The same damage to the elastic fibers was also present in an additional specimen from grossly uninvolved skin. On electron microscopy there were pronounced changes of the elastic fibers with elastolysis in both specimens. This case with generalized damage of the dermal elastic tissue supports the view that elastorrhexis is the key feature of AKE. Accordingly, the disease could be regarded as a primary elastic tissue disorder. The marginal acral keratoderma that is seen in AKE patients could represent epidermal changes secondary to chronic trauma. PMID- 9870681 TI - Functioning human insulinomas. An immunohistochemical analysis of intracellular insulin processing. AB - Sixty-seven insulinomas were investigated by immunohistochemistry using site directed antibodies against insulin, proinsulin, chromogranin A, HISL-19, and four proteins directly or indirectly involved in the proteolytic processing of proinsulin: the prohormone convertases PC2 and PC3, carboxypeptidase H (CPH) and 7B2. Results were expressed in a six-grade score according to the frequency of immunoreactive tumour cells. Insulin was expressed by all tumours, appearing in either a diffuse or a polarized pattern and being detected in more than 30% of tumour cells in all cases but three. Proinsulin was also expressed in all tumours, with more than 50% of tumour cells immunoreactive in all cases but 5. It was consistently localized in the Golgi apparatus. In about half the cases, moreover, it also showed diffuse cytoplasmic staining, usually with a very sparse distribution. Trabecular and solid insulinomas did not present specific, homogeneous patterns of insulin immunostaining. However, insulin immunoreactivity was much more abundant in trabecular than in solid neoplasms, being present in virtually all tumour cells (score 6) in 50% and 8% of cases, respectively. Virtually all insulinomas expressed PC2, PC3, CPH and 7B2, usually in 30-100% of tumour cells, with a frequency significantly related to that of insulin. However, detection of PC2 and 7B2 was slightly less frequent than that of PC3 and CPH. In consecutive sections these proteins were found to be mostly co-localized with insulin and chromogranin A but not with proinsulin. They were heavily expressed in all 10 tumours with more than 10% of cells showing cytoplasmic proinsulin immunoreactivity, indicating that the leakage of proinsulin from the Golgi compartment is not associated with faulty expression of converting enzymes and possibly reflects a saturated processing capacity. HISL-19 immunoreactivity was found in both Golgi apparatus and insulin stores, indicating that the relevant antigen is different from all other proteins investigated. These results do not support a defect in expression or localization of proinsulin-processing enzymes in most insulinomas. PMID- 9870682 TI - Bilateral apocrine carcinoma of the breast. Molecular and immunocytochemical evidence for two independent primary tumours. AB - Apocrine carcinoma is an uncommon variant of breast cancer. The frequency of bilaterality in patients who have apocrine carcinoma in one breast is not significantly different from that for bilateral mammary carcinomas in general, but bilateral apocrine carcinomas are very uncommon. We report on a bilateral apocrine carcinoma of the breast in a 74-year-old woman. The apocrine differentiation in both tumours was confirmed by the positivity of the cytoplasmic granules for PAS after diastase digestion and immunoreactivity for GCDFP-15 and sialyl-Tn. The tumour in the right breast showed immunohistochemical expression of p53, and a mutation was demonstrated by PCR-SSCP; the tumour in the left breast was negative for p53 on immunohistochemistry, and no mutation was found at the molecular level. c-erbB2 expression was not detected in the right tumour but there was overexpression (at the cell membrane) in the left tumour. Both tumours were aneuploid: the right tumour displayed multiple stemlines, whereas the left tumour had a triploid profile. Using the fluorescence in situ hybridization technique we demonstrated that both tumours displayed chromosome 17 polysomy and numerical abnormalities of chromosome 1, polysomy in the right and monosomy in the left tumour. We conclude that the two tumours are probably independent, as are most bilateral carcinomas of the breast. PMID- 9870683 TI - Overexpression of c-met proto-oncogene associated with chromophilic renal cell carcinoma with papillary growth. AB - Various genetic changes are involved in human renal cell carcinomas (RCCs). However, the molecular events related to other cytomorphological subtypes of RCC are not well known, apart from the relationship between the von Hippel-Lindau tumour suppressor gene and clear cell subtype RCC. We examined the overexpression of several growth factor receptors immunohistochemically and analyzed their relationship to the cytomorphological characters in 120 cases of RCCs. These receptors included c-met proto-oncogene product (c-MET), epidermal growth factor receptor (EGFR) and transforming growth factor beta receptor II (TGFbetaR). The overexpression of c-MET was detected in all cases (20/20) of the tubulo-papillary growth type and 78.3% (18/23) of chromophilic cell subtype, resulting in a very significant associations between c-MET overexpression and tubulo-papillary growth RCCs (P<0.0001), c-MET and chromophilic subtype RCCs (P<0.0001), and c-MET and EGFR (P<0.0001). EGFR overexpression was significantly associated with the compact growth RCCs (49/89, P<0.0001), clear cell subtype RCCs (P<0.005) and the overexpression of TGFbetaR (P<0.0001). These results strongly suggest a close correlation between the overexpression of c-MET and development of the chromophilic subtype of RCC with papillary growth pattern. EGFR expression is closely related to the pathogenesis of the clear cell subtype of RCC with compact growth pattern. The overexpression of c-MET, EGFR, and TGFbetaR may have roles that are individually significant in the morphogenesis of RCC. PMID- 9870684 TI - The fibrotic focus in advanced colorectal carcinoma: a hitherto unrecognized histological predictor for liver metastasis. AB - A fibrotic focus (FF) is a clearly defined area consisting of fibroblasts and/or collagen fibres arranged in irregular or storiform patterns within tumours. We looked to see whether FF in advanced colorectal carcinoma was associated with distant organ metastasis especially to the liver. The correlation between FF and the presence of synchronous or total (synchronous and/or metachronous) liver metastasis and tumour recurrence was assessed in 77 patients with Dukes B and C advanced colorectal carcinoma treated by resection. The median follow-up period was 21 months. In multivariate analysis, FF significantly increased the relative risk (RR) of synchronous liver metastasis (RR=4.9, P<0.05) and total liver metastasis (RR=4.6, P<0.05). FF also increased the RR of tumour recurrence (RR=2.4), but the increase was not statistically significant. FF is a newly recognized histological indicator of liver metastasis in advanced colorectal carcinoma. PMID- 9870685 TI - Imbalance between proliferation and apoptosis in the development of colorectal carcinoma. AB - To evaluate the relationship between cell proliferation and apoptosis in sporadic colorectal carcinogenesis, immunohistochemistry for proliferation-associated antigen Ki-67 and in situ end labelling for identifying apoptotic bodies were performed on paraffin sections from 59 adenomas and 22 carcinomas. These results were correlated with the expression of the proliferation and apoptosis modulators Bcl-2 and p53. Carcinomas showed increased proliferation and apoptosis compared with adenomas (P<0.0001, P<0.001, respectively). There were positive linear correlations between proliferation and apoptosis in adenomas and carcinomas (P<0.02, P<0.05, respectively). The proliferative rate increased significantly from mild to moderate, and from moderate to severe dysplasia (P<0.002, P<0.001, respectively). Apoptotic rate also increased in this sequence, but the increases did not reach statistical significance (both P>0.05). Expression of Bcl-2 was associated with lower apoptotic rate in adenomas (P<0.025) but not in carcinomas (P>0.25), whereas p53 expression was correlated with higher proliferative rate in both adenomas and carcinomas (P<0.01, P<0.05, respectively). An inverse relationship between Bcl-2 and p53 expression was seen in both adenomas and carcinomas (P<0.05, P<0.005, respectively). These data suggest that the normal balance between proliferation and apoptosis is disturbed in colorectal carcinogenesis, both being increased, but proliferation occurs in excess. Bcl-2 and p53 may each play a role in modulating cell apoptosis or proliferation during the development of colorectal carcinoma. PMID- 9870686 TI - Immunohistochemical detection of human mtDNA polymerase gamma and of human mitochondrial transcription factor A in cytochrome-c-oxidase-deficient oxyphil cells of hyperfunctional parathyroids. AB - Immunohistochemical studies were performed in 18 hyperfunctional parathyroids with oxyphil cell aggregates for the detection of cytochrome-c-oxidase (complex IV of the respiratory chain), mitochondrial DNA polymerase gamma and human mitochondrial transcription factor A (h-mtTFA). Seventy-three oxyphil areas exhibiting a defect of cytochrome-c-oxidase were found. The defect involved both the mitochondrially coded subunits II/III and the nuclear derived subunits Vab. There was no loss of mtDNA polymerase gamma or of h-mtTFA in these foci, corresponding to a high content of mtDNA revealed by in situ hybridization. Isolated defects of h-mtTFA were also not found. In contrast, isolated defects of mtDNA polymerase gamma were present in 22 oxyphil foci. These results show that defects of cytochrome-c-oxidase in oxyphil cells are not due to altered expression of h-mtTFA or DNA polymerase gamma, indicating that other nuclear factors involved in the generation of the respiratory chain may be impaired. The low incidence of defects of mtDNA polymerase gamma and the absence of alterations of h-mtTFA and cytochrome-c-oxidase in these foci suggest that defects of mtDNA polymerase gamma are of minor pathogenetic significance. PMID- 9870688 TI - Intranuclear iron deposition in hepatocytes and renal tubular cells in mice treated with ferric nitrilotriacetate. AB - Cytoplasmic and intranuclear iron depositions were observed in the livers and kidneys of male and female ddY mice treated for 4-12 weeks with ferric nitrilotriacetate (Fe-NTA), a known renal carcinogen that acts through the production of free radicals. The intranuclear iron deposition consisted of a spherical aggregation of ferritin particles of approximately 10 nm diameter, as revealed by immunohistochemical staining and electron microscopy. Although the incidence of renal tumours was greater in the males than in the females, the incidence of iron depositions did not differ with gender. The most abundant intranuclear iron depositions were observed in the animals treated with Fe-NTA for the longest duration (12 weeks). These findings suggest that the intranuclear production and propagation of free radical reactions are prevented by the trapping of iron in a chemically inert iron form of ferritin. PMID- 9870687 TI - Albumin gene expression in adenocarcinomas with hepatoid differentiation. AB - Two cases of hepatoid adenocarcinomas were studied with an in situ hybridization technique (ISH) using a RNA probe for human albumin mRNA. In case 1 the urinary bladder of a 67-year-old woman was affected; in case 2 the tumour was located in the gastric antrum of an 80-year-old woman. In neither case had alpha fetoprotein (AFP) been determined preoperatively. Histologically these cases showed adenocarcinomatous features intermingled with hepatoid areas. These latter areas were characterized by cords of polygonal cells, each with an oval nucleus and prominent nucleoli, separated by a fine network of sinusoids. In the hepatoid areas the immunohistochemical profile was similar to that observed in hepatocellular carcinomas, in that the tumour cells were positive with AFP, alpha 1-antitrypsin (A1AAT) and albumin antisera and there was a canalicular type of reactivity with polyclonal anti-CEA (pCEA) antibody. ISH revealed albumin mRNA in virtually all hepatoid cells in case 1, and in about 50% of those in case 2. In addition, in case 2 occasional cells in the adenocarcinomatous areas showed albumin transcripts of ISH. Our findings confirm that ISH for albumin mRNA probe is a valuable method of establishing hepatocellular differentiation, and that hepatoid adenocarcinomas are tumours with true extrahepatic hepatocellular differentiation. PMID- 9870689 TI - The contribution of nitric oxide to renal vascular wall thickening in rats with L NAME-induced hypertension. AB - We investigated the mechanisms of renal vascular wall thickening in a rat model of N-nitro L-arginine methyl ester (L-NAME)-induced hypertension. To separate the effects of L-NAME-induced hypertension from other effects of nitric oxide (NO) inhibition, we created two models of L-NAME-induced hypertension: both had the same blood pressure level but NO inhibition was moderate in one group (group M) and severe in the other (group S). Urinary excretion of nitrates and nitrites was lower in group S than in group M. Wall thickening and lipid deposition in renal vessels were significantly greater in group S than in groups M. Simple and multiple regression analyses indicated that renal vascular wall thickening was more strongly correlated with lipid deposition than with blood pressure. The number of vessels positive for staining with Sudan black B was negatively correlated with urinary NO excretion. Expression of fibronectin and transforming growth factor-beta was greater in the Sudan black B-positive than in the Sudan black B-negative vessels, suggesting that extracellular matrix production was increased in vessels with lipid deposition. Lipid deposition and increased production of extracellular matrix may contribute to renal vascular wall thickening in L-NAME-induced hypertension. Some mechanisms independent of hypertension play important roles in vascular wall thickening induced by NO inhibition. PMID- 9870690 TI - Observations on bone formation and remodelling in advanced atherosclerotic lesions of human carotid arteries. AB - Immunolocalisation and histochemical techniques were used to examine mineralised bone deposits within late stage atherosclerotic plaques of human carotid arteries. These specimens showed characteristic features of osteogenesis. Large calcifications were often observed in close association with or integrated within mineralised bone. Smooth muscle cells (alpha-actin positive) were often located around osteoid-like matrix, together with focal accumulations of macrophages (CD68 and HAM56 positive). Local accumulations of mast cells (tryptase positive) were consistently observed in close association with the bone. Multinucleated giant cells in close apposition with mineralised bone demonstrated typical osteoclastic morphology, and were positively stained for acid phosphatase and the macrophage marker CD68. Thus, all the normal features of bone formation and resorption were observed in this microcosm of osteogenesis within atherosclerotic plaque; the term 'osteosome' seems appropriate for the structure. These osteosomes have numerous advantages for experimental studies of the various osteogenic factors responsible for bone metabolism, especially following short term tissue culture. This ex vivo technique was used to demonstrate the distribution and the multiple cellular sources of bone morphogenetic protein 6. PMID- 9870691 TI - Localization of vascular endothelial growth factor in synovial membrane mast cells: examination with "multi-labelling subtraction immunostaining". AB - Mast cells are believed to play a novel part in the development of destructive synovial pannus in rheumatoid arthritis (RA). This study was undertaken to investigate the localization of vascular endothelial growth factor (VEGF) in the synovial membrane using a unique immunostaining technique. Synovial specimens of RA patients were examined immunohistochemically and were compared with specimens from non-RA controls. Multi-labelling subtraction immunostaining, a modification of double- and triple-labelling immunostaining, revealed that the VEGF-positive cells were identical to tryptase-positive cells (mast cells). No other cell types were found to be positive for VEGF. The synovium of RA patients showed a larger number of VEGF-positive mast cells than that of non-RA controls (P<0.001). The study suggests that mast cell-derived VEGF may contribute to the development of synovial pannus in RA. PMID- 9870692 TI - Intracranial germ cell tumour (embryonal carcinoma with teratoma) with complex karyotype including isochromosome 12p. AB - We report the chromosomal characteristics of a malignant teratoma with embryonal carcinoma component located in the pineal region of a 15-year-old boy. Chromosome analysis showed a near-triploid complex karyotype (62 chromosomes), including two copies of an isochromosome 12p, confirmed by fluorescence in situ hybridization analysis. The present findings indicate that isochromosome 12p formation is probably associated with the development of malignant germ cell tumours. PMID- 9870693 TI - Mucolipidosis type II with evidence of a novel storage site. AB - In a case of infantile mucolipidosis type II (I-cell disease), storage was identified at autopsy in serous-type secretory cells in exocrine pancreas, in the tracheal and sublingual salivary glands and in the chief (zymogenic) cells of the gastric oxyntic glands, suggesting a systemic involvement of this type of secretory cells. The content of specific secretory granules was inversely proportional to the intensity of the storage process. The mucus-producing cells were not affected. The serous glandular system is a novel storage site in I-cell disease. Review of archival material in three further cases confirmed the findings. PMID- 9870694 TI - A child with a t(11;19)(q14-21;p12) in a pulmonary mucoepidermoid carcinoma. AB - We report on a mucoepidermoid carcinoma (MEC) of the lung in a 6-year-old girl with a t(11;19)(q14-21;p12) as the sole karyotypic abnormality. An apparently identical t(11;19) has been reported previously in a MEC originating from the major and minor salivary glands. Our findings indicate that the t(11;19) is intimately associated with the mucoepidermoid phenotype and may be used as a diagnostic marker for this tumour type. PMID- 9870695 TI - Gastric carcinoma risk index in patients infected with Helicobacter pylori. PMID- 9870696 TI - Instability at the k2 Mdh1-n y20 chromosomal region in soybean. AB - Ten mutants have been reported at the k2 (tan saddle seed coat) Mdh1-n (mitochondrial malate dehydrogenase 1 null) y20 (yellow foliage) chromosomal region in soybean [Glycine max (L.) Merr.]. The precise genetic mechanism(s) responsible for generating these mutants is (are) not known. The objective of this study was to determine whether chromosomal instability exists at this region. We introduced the w4-m and Y18-m mutable systems into the three independent sources of tan saddle seed coat mutants, T239 (k2), T261 (k2 Mdh1-n), and L67-3483 (k2). A total of 12 bright yellow mutants were isolated with tan saddle seed coat, malate dehydrogenase 1 null phenotypes. Of these, 11 were found in 11 F2 mutant families out of a total of 977 derived by crossing T239 (k2), T261 (k2 Mdh1-n), and L67-3483 (k2) with six lines suspected to contain active transposable elements. One was found in the F3 generation derived from the cross A1937 x T239 (k2). Of the 11 F2 mutant families, 10 (out of a total of 381 F2 families) were associated with the T239 (k2) genetic background, and one out of 323 was associated with the T261 (k2 Mdh1-n) genetic background. But no mutation events were found among the 273 families with the L67-3483 (k2) genetic background. Allelism and inheritance studies indicated that these 12 bright yellow mutants were new mutants in the k2 Mdh1-n y20 chromosomal region. Thus, on introducing the w4-m and Y18-m mutable systems into T239 (k2) and T261 (k2 Mdh1 n) genetic backgrounds, chromosomal instability was induced in this region. In addition, 21 greenish yellow mutants were identified in the total of 977 F2 families. All 21 greenish yellow mutants were associated with the T239 (k2) genetic background. The mutations for greenish yellow foliage affected foliage color only at the seedling stage. Cosegregation of the tan saddle seed coat character with greenish yellow foliage were observed for these 21 greenish yellow mutants, suggesting that the greenish yellow phenotype may be due to a pleiotropic effect of the k2 allele in T239 or to chromosomal rearrangements at or near the k2 allele in T239. Finally, we believe that the genetic mechanism responsible for this high frequency of instability at the k2 Mdh1-n y20 chromosomal region involves receptor element activities present at this chromosomal region, which may contain complex chromosomal rearrangements in T239 and T261. PMID- 9870697 TI - Differential effects of caffeine on DNA damage and replication cell cycle checkpoints in the fission yeast Schizosaccharomyces pombe. AB - Caffeine potentiates the lethal effects of ultraviolet and ionising radiation on wild-type Schizosaccharomyces pombe cells. In previous studies this was attributed to the inhibition by caffeine of a novel DNA repair pathway in S. pombe that was absent in the budding yeast Saccharomyces cerevisiae. Studies with radiation-sensitive S. pombe mutants suggested that this caffeine-sensitive pathway could repair ultraviolet radiation damage in the absence of nucleotide excision repair. The alternative pathway was thought to be recombinational and to operate in the G2 phase of the cell cycle. However, in this study we show that cells held in G1 of the cell cycle can remove ultraviolet-induced lesions in the absence of nucleotide excision repair. We also show that recombination-defective mutants, and those now known to define the alternative repair pathway, still exhibit the caffeine effect. Our observations suggest that the basis of the caffeine effect is not due to direct inhibition of recombinational repair. The mutants originally thought to be involved in a caffeine-sensitive recombinational repair process are now known to be defective in arresting the cell cycle in S and/or G2 following DNA damage or incomplete replication. The gene products may also have an additional role in a DNA repair or damage tolerance pathway. The effect of caffeine could, therefore, be due to interference with DNA damage checkpoints, or inhibition of the DNA damage repair/tolerance pathway. Using a combination of flow cytometric analysis, mitotic index analysis and fluorescence microscopy we show that caffeine interferes with intra-S phase and G2 DNA damage checkpoints, overcoming cell cycle delays associated with damaged DNA. In contrast, caffeine has no effect on the DNA replication S phase checkpoint in response to inhibition of DNA synthesis by hydroxyurea. PMID- 9870698 TI - The genes encoding light-harvesting subunits of Cyclotella cryptica (Bacillariophyceae) constitute a complex and heterogeneous family. AB - Total RNA was isolated from the diatom Cyclotella cryptica and separated into poly(A)+ and poly(A)- fractions. These fractions were subjected to in vitro translation/immunoprecipitation experiments using an antiserum directed against the predominant light-harvesting complex of Cy. cryptica (ccry antiserum) and a heterologous antiserum raised against the light-harvesting complex of the cryptophyte Cryptomonas maculata (cmac antiserum). From translation reactions programmed with poly(A)+ RNA the ccry-antiserum immunoprecipitated polypeptides with relative molecular weights (Mr) of 27000, 25000, 23000 and 21000, while the cmac-antiserum precipitated proteins with Mrs of 32500 and 27000, respectively. Subsequent cDNA synthesis and immunological screening of the cDNA library with both antisera resulted in the isolation of six cDNA clones encoding light harvesting subunits. Full-length precursors were 199-210 amino acids in length and had Mrs of 20000-23000. The lengths of the putative signal peptides were 29 or 30 amino acids. Pairwise comparison revealed that the similarity between the clones ranged from 54-99% on the nucleotide level and from 36-99% at the amino acid level. In agreement with the data from the screens with the two antisera, the genes clustered into two groups. The data provide evidence that the genes constitute a heterogeneous multigene family and that the light-harvesting system of Cy. cryptica might be as complex as that of higher plants and green algae. PMID- 9870699 TI - A second gene for type I signal peptidase in Bradyrhizobium japonicum, sipF, is located near genes involved in RNA processing and cell division. AB - The TnphoA-induced Bradyrhizobium japonicum mutant 184 shows slow growth and aberrant colonization of soybean nodules. Using a DNA fragment adjacent to the transposon insertion site as a probe, a 3.4-kb BglII fragment of B. japonicum 110spc4 DNA was identified and cloned. Sequence analysis indicated that two truncated ORFs and three complete ORFs were encoded on this fragment. A database search revealed homologies to several other prokaryotic proteins: PdxJ (an enzyme involved in vitamin B6 biosynthesis), AcpS (acyl carrier protein synthase), Lep or Sip (prokaryotic type I signal peptidase), RNase III (an endoribonuclease which processes double-stranded rRNA precursors and mRNA) and Era (a GTP-binding protein required for cell division). The mutation in strain 184 was found to lie within the signal peptidase gene, which was designated sipF. Therefore, sipF is located in a region that encodes gene products involved in posttranscriptional and posttranslational processing processes. By complementation of the lep(ts) E. coli mutant strain IT41 it was demonstrated that sipF indeed encodes a functional signal peptidase, and genetic complementation of B. japonicum mutant 184 by a 2.8 kb SalI fragment indicated that sipF is expressed from a promoter located directly upstream of sipF. Using a non-polar kanamycin resistance cassette, a specific sipF mutant was constructed which exhibited defects in symbiosis similar to those of the original mutant 184. PMID- 9870700 TI - Recombination between chloroplast genomes of Trachystoma ballii and Brassica juncea following protoplast fusion. AB - We document here the presence of a recombinant plastome in a cytoplasmic male sterile (CMS) line of Brassica juncea developed from the somatic hybrid Trachystoma ballii + B. juncea. Restriction endonuclease digestion of the chloroplast (cp) DNA has revealed that the recombinant plastome gives rise to novel fragments in addition to the parent-specific fragments. Analysis of the 16S rRNA region by Southern hybridization shows no variation between B. juncea, T. ballii and the CMS line. The rbcL gene region of the recombinant plastome is identical to that in T. ballii. Analysis with probes for psbA and psbD using single and double DNA digests indicates that the hybridization patterns of the recombinant plastome are identical to those of the parents in digests obtained with some restriction enzymes, while novel bands hybridize to probes in other digests. In the psbA region, a B. juncea-specific PstI site and a T. ballii specific EcoRI site are found in the recombinant plastome. The pshD region of the recombinant plastome contains a B. juncea-specific HindIII site and T. ballii specific BamHI and HpaII sites. These results indicate the occurrence of intergenomic recombination between the chloroplasts of T. ballii and B. juncea in the somatic hybrid from which the CMS line was developed. The recombined plastome appears to be a mosaic of fragments specific to both parents and the recombination event has occurred in the single-copy regions. These recombinational events have not caused any imbalance in the recombinant plastome in terms of chloroplast-related functions, which have remained stable over generations. PMID- 9870701 TI - Effect of ploidy and homozygosity on transgene expression in primary tobacco transformants and their androgenetic progenies. AB - Expression of a transgene is rarely analysed in the androgenetic progenies of the transgenic plants. Here, we report differential transgene expression in androgenetic haploid and doubled haploid (DH) tobacco plants as compared to the diploid parental lines, thus demonstrating a gene dosage effect. Using Agrobacterium-mediated transformation, and bacterial reporter genes encoding neomycin phosphotransferase (nptII) and beta-glucuronidase (uidA/ GUS), driven respectively by the mas 1' and mas 2' promoters, we have generated more than 150 independent transgenic (R0) Nicotiana tabacum plants containing one or more T-DNA copies. Transgene analyses of these R0, their selfed R1 lines and their corresponding haploid progenies showed an obvious position effect (site of T-DNA insertion on chromosome) on uidA expression. However, transgene (GUS) expression levels were not proportional to transgene copy number. More than 150 haploids and doubled haploids, induced by treatment with colchicine, were produced from 20 independent transgenic R0 plants containing single and multiple copies of the uidA gene. We observed that homozygous DH plants expressed GUS at approximately 2.9-fold the level of the corresponding parental haploid plants. This increase in transgene expression may be attributed mainly to the increase (2-fold) in chromosome number. Based on this observation, we suggest a strong link between chromosome number (ploidy dosage effect) and transgene expression. In particular, we demonstrate the effect on its expression level of converting the transgene from the heterozygous (in R0 plants) to the homozygous (DH) state: e.g. an increase of 50% was observed in the homozygous DH as compared to the original heterozygous diploid plants. We propose that ploidy coupled with homozygosity can result in a new type of gene activation, creating differences in gene expression patterns. PMID- 9870702 TI - A maize Myb homolog is encoded by a multicopy gene complex. AB - The maize P gene encodes a Myb-homologous transcriptional regulator of flavonoid pigmentation in floral organs, and different P gene alleles condition precise tissue- and organ-specific pigmentation patterns. To determine the molecular basis for allele-specific expression patterns, we have isolated and compared two natural alleles of the P gene which differ in expression, structure and copy number. The P-rr allele is associated with pigmentation of most floral tissues and contains a single copy of the P gene. In contrast, the P-wr allele restricts pigmentation to a subset of floral tissues, and is composed of six gene copies arranged in a tandem head-to-tail array. Each of the six repeats contains a single P gene, including regulatory and coding sequences. Despite the six-fold tandem repetition of P-wr gene copies, P-wr mRNA levels in kernel pericarp are much reduced compared to mRNA levels from the single-copy P-rr gene. Moreover, the P-wr multicopy complex is hypermethylated relative to P-rr. Thus, maize P gene alleles may represent a natural system for studying the effects of methylation and gene copy number on tissue-specific gene expression. We discuss the possibility that somatic pairing of repeated gene copies may be involved in regulating gene expression. PMID- 9870703 TI - Insertion specificity and trans-activation of IS801. AB - The transposable element IS801, isolated from plasmid pMMC7105 of Pseudomonas syringae pv. phaseolicola, transposes in Escherichia coli to plasmid targets, expressing a relatively relaxed target specificity. The target sequences are tetramers with homology with the left terminus (GAAC) of the transposing unit, the alternative targets being GAAC, GGAC, CAAG, and CGAC. In the areas flanking IS801 in 13 different locations, no similarities other than the target tetramer were observed. The transposase is physically and functionally separable from the transposing unit since transposition of constructs carrying marker genes occurs with the transposase expressed in trans. The IS801 transposase shows amino acid sequence homology to the transposases of the E. coli elements IS91 and IS1294. These transposases contain conserved amino acid motifs found in the replicases of certain plasmids that replicate as rolling circles. PMID- 9870704 TI - Rice has two distinct classes of protein kinase genes related to SNF1 of Saccharomyces cerevisiae, which are differently regulated in early seed development. AB - We have isolated five cDNA clones (osk1-5) for protein kinases from rice which are related to SNF1 protein kinase of Saccharomyces cerevisiae. Based on the sequence homology, these cDNAs can be classified into two groups, group 1 (osk1) and group 2 (osk2-5). The products of these genes were demonstrated to be functional SNF1-related protein kinases by in vitro and in vivo experiments. Recombinant proteins expressed from both groups of genes were fully active as protein kinases and could phosphorylate SAMS peptide, a substrate specific for the SNF1/AMPK family, as well as themselves (autophosphorylation). Moreover, expression of osk3 cDNA in yeast snf1 mutants restored SNF1 function. Northern blot analyses showed differential expression of these two gene groups; group 1 is expressed uniformly in growing tissues (young roots, young shoots, flowers, and immature seeds), whereas group 2 is strongly expressed in immature seeds. SNF1 related protein kinases have been reported from different plant species, such as rye, barley, Arabidopsis, tobacco, and potato, while the type of gene strongly expressed in immature seeds is known only in cereals such as rye, barley, and, from our findings, in rice. Expression levels of the group 2 genes were further analyzed in seeds during seed maturation. Expression is transiently increased in the early stages of seed maturation and then decreases. The expression peak precedes those of the sbe1 and waxy genes, which are involved in starch synthesis in rice. Taken together, these findings suggest that group 2 OSK genes play important roles in the early stages of endosperm development in rice seeds. PMID- 9870705 TI - Replication-dependent and selection-induced mutations in respiration-competent and respiration-deficient strains of Saccharomyces cerevisiae. AB - Adaptive or selection-induced mutations are defined as mutations that occur in non-dividing cells as a response to prolonged non-lethal selective pressure such as starvation for an essential amino acid. In the absence of DNA replication, the processing of endogenous DNA lesions by repair enzymes probably acts as a source of mutations. We are studying selection-induced reversions of frameshift alleles in the eukaryote Saccharomyces cerevisiae. Here we show that respiration deficient strains, totally devoid of mitochondrial DNA, yield selection-induced mutants at slightly elevated frequencies compared to isonucleic respiration competent strains. Therefore factors of mitochondrial origin such as reactive oxygen species or hypothetical recombinogenic DNA fragments are unlikely to be mediators of selection-induced nuclear frameshift mutation in yeast. Furthermore we compared sequence spectra of reversions of the +1 hom3-10 frameshift allele and found a strong preference for -1 deletions in mononucleotide repeats in selection-induced and replication-dependent revertants, indicating slippage errors during DNA repair synthesis as well as during DNA replication. Remarkably, a higher degree of variation in the site of the reverting frameshift and accompanying base substitutions was found among selection-induced revertants. PMID- 9870706 TI - Cell survival effect of activin against heat shock stress on OVCAR3. AB - Activin has been known as the hormone protein which regulates either cell proliferation or cell differentiation. Recently, it has also been reported that activin may have cell survival function. In this study, we have investigated, 1) the expression of inhibin subunits and activin receptors (ActRs) in ovarian carcinoma cell line (OVCAR3), 2) the binding property between activin and its receptors under the exposure to stress, and 3) the effect of activin on cell proliferation. All of inhibin subunits and ActR Ia, IIa and IIb mRNA were amplified by RT-PCR in OVCAR3. By Western blot analysis, ActR IIa and IIb proteins were detected. The binding property between activin and ActRs was analyzed with the fixed complex, using chemical cross linker. The bigger molecular weight signals, which had been shown to form the heterotrimeric complex among activin, ActR type I and ActR type II were detected after cross linking. These upper signals were apparently increased by rh-Activin and decreased by rh Follistatin. Therefore, it was suggested that they were resultant from activin and Act-R complex. OVCAR3 was exposed to the stress (42C, 1 hour heat shock), the protein level of ActR IIa increased and ActR IIb decreased from about 3 h to 24 h after the exposure to the heat stress (HS). On the other hand, the complex between activin and ActR IIa and IIb increased from 3 h after the exposure to HS. To investigate the effect of activin and follistatin on OVCAR3 proliferation after the exposure to HS, we counted the cell number at 96 h after the treatment with activin or follistatin in the condition either with or without HS. Proliferation of the cell in the presence of HS was stimulated by rh-Activin and inhibited by rh-Follistatin. These data suggest that activin might have the function to survive and to proliferate OVCAR3, due to, at least in part the increase in its binding capacity to ActRs through either autocrine or paracrine manner. PMID- 9870707 TI - Effect of sodium thiopentone anesthesia on the phagocytic activity of rat peritoneal macrophages. AB - To elucidate the effect of sodium thiopentone anesthesia on the function of phagocytic cells, albino rats were anesthetized with 60 mg/kg. of sodium thiopentone. After 90 min., peritoneal macrophages were harvested and their capacity for superoxide anion generation was detected. Following anesthesia for 90 min. latex particles were injected intraperitoneally, and after additional 30 min. the macrophages were derived, embedded in agar and the number of cells engaged in phagocytosis, as well as the number of latex particles engulfed by each individual cell were counted in semi-thick sections. Macrophages of anesthetized animals showed a statistically significant decrease of both superoxide anion generation and mean number of phagocytic cells, and engulfed fewer particles than those of the controls. Similar results were obtained following incubation of the cells with sodium thiopentone in vitro. The serum corticosterone level in anesthetized rats was significantly higher than that of the control animals. The results indicate that impaired phagocytosis following anesthesia induced by sodium thiopentone, in addition to alterations of the immune system caused by surgical trauma, may be one of the reasons for increased susceptibility to infections of surgical patients during the postoperative period. PMID- 9870708 TI - The anxiolytic effect on the fear-potentiated startle is not due to a non specific disruption. AB - In the present study, the effects of alprazolam, chlordiazepoxide, ethanol and haloperidol in the strychnine-potentiated startle response paradigm were investigated. Because strychnine increases control startle levels without fear conditioning, no central state of fear exists. When anxiolytic drugs reduce the fear-induced potentiation of the startle response without reducing the strychnine induced startle potentiation, their attenuating effect on startle potentiation in the fear-potentiated startle response paradigm can more likely be attributed to their anxiolytic properties. The selected drugs, which effectively reduced the fear-induced startle potentiation in an earlier study, indeed did not selectively affect the strychnine-induced startle potentiation, supporting the notion that the dose-dependent reduction of startle potentiation due to anxiolytic drugs in the fear-potentiated startle response paradigm does not reflect a "non-specific" disruption of startle behavior. PMID- 9870709 TI - Interaction between arginine vasopressin- and raised extracellular potassium stimulated pathways in adrenocorticotropin secretion. AB - The intracellular regulation of adrenocorticotropin (ACTH) secretion from pituitary corticotroph cells was investigated by simultaneously exposing cultured ovine corticotrophs to arginine vasopressin (AVP) and raised extracellular K+ concentration ([K+]e). Both of these secretagogues activate L-type voltage sensitive calcium channels (L-VSCC) as part of their respective ACTH secretory responses. When given together at high concentrations, AVP and raised [K+]e caused ACTH responses that were smaller in magnitude than the sum of the individual responses. However, at low agonist concentrations the simultaneous responses were greater in magnitude (i.e., synergistic). Further investigation suggested that activation of protein kinase C (PKC), which is part of the AVP induced intracellular signalling pathway, is necessary and sufficient for the generation of the synergistic response, although it is not obligatory for AVP induced ACTH release. PMID- 9870710 TI - Changes in the liver protein pattern of female Wistar rats treated with the hypoglycemic agent SDZ PGU 693. AB - SDZ PGU 693 acts as a hypoglycemic agent by stimulating glucose utilisation in insulin-sensitive peripheral tissues, such as skeletal muscle and fat. In a 28 day toxicity study the compound was found to induce hepatocellular hypertrophy in Wistar rats treated with 300 mg/kg/day. To gain insights into the pathomechanism of these alterations, aliquots of liver samples from control and treated female Wistar rats were separated by two-dimensional protein gel electrophoresis and the digitized images of the protein patterns were searched for protein abundance changes. Significant treatment-related quantitative changes (P < 0.001) were found in 29 liver proteins. Major increases were observed in several microsomal proteins, including NADPH cytochrome P-450 reductase, cytochrome b5 and serine protease inhibitor. The changes in the cytochrome related enzymes, both known co factors of the P-450 enzyme system, strongly suggest that SDZ PGU 693 induces microsomal proliferation and induction of the P-450 enzyme system. Decreases were observed in a series of mitochondrial proteins, such as F1ATPase-delta subunit and ornithine aminotransferase precursor as well as in several cytosolic proteins such as the liver fatty acid binding protein, arylsulfotransferase and the senescence marker protein-30. The changes in F1ATPase-delta subunit and liver fatty acid binding protein together suggest a down-regulation of the mitochondrial liver fatty acid metabolism, likely reflecting the pharmacological action of the compound. These results show that SDZ PGU 693 produces a complex pattern of gene expression changes which give insights into the molecular mechanisms of both its pharmacological action and a toxic response. PMID- 9870711 TI - Expression of prolactin messenger ribonucleic acid in the mouse gonads during sexual maturation. AB - The pituitary hormone prolactin is known to be produced in various extrapituitary tissues as well. We examined the presence of prolactin mRNA in the mouse gonads by reverse transcription (RT)-PCR and nucleotide sequence analysis. Both ovary and testis were found to express small amounts of mRNA coding prolactin, identical to that in the pituitary gland. Next, we established a sensitive competitive RT-PCR method to estimate the amount of prolactin mRNA and then measured its expression in the gonads during sexual maturation (10-80 days of age). In the ovary, the amount of prolactin mRNA (copies/microg of total RNA) gradually decreased from day 10 to 40, and the lower level was maintained until day 80. Little difference in the amount was observed between the estrous and diestrous groups on day 80. Conversely, the amount of testicular prolactin mRNA gradually increased from day 10 to 80. These results suggest that prolactin is produced in the mouse gonads and that it acts as an autocrine/paracrine factor modulating gonadal functions. PMID- 9870712 TI - Ursodeoxycholate protects against ethanol-induced liver mitochondrial injury. AB - The purpose of this work was to examine whether ursodeoxycholate (UDC), a hydrophilic bile salt, could reduce mitochondrial liver injury from chronic ethanol consumption in rats. Animals were pair-fed liquid diets containing 36% of calories as ethanol or isocaloric carbohydrates. They were randomly assigned into 4 groups of 7 rats each and received a specific treatment for 5 weeks: control diet, ethanol diet, control diet + UDC, and ethanol diet + UDC. Respiratory rates of isolated liver mitochondria were measured using a Clark oxygen electrode with sodium succinate as substrate. Mitochondria from rats chronically fed ethanol demonstrated an impaired ability to produce energy. At the fatty liver stage, the ADP-stimulated respiration (V3) was depressed by 33%, the respiratory control ratio (RC) by 25% and the P/O ratio by 15%. In ethanol-fed rats supplemented with UDC, both the rate and efficiency of ATP synthesis via the oxidative phosphorylation were improved: V3 was increased by 35%, P/O by 8%. All the respiratory parameters were similar in control group and control + UDC group. On the other hand, the number and size of mitochondria were assessed by electron microscopy and computer-assisted quantitative analysis. The number of mitochondria from ethanol-treated rats was decreased by 29%, and they were enlarged by 74%. Both parameters were normalized to control values by UDC treatment. These studies demonstrate that UDC has a protective effect against ethanol-induced mitochondrial injury by improving ATP synthesis and preserving liver mitochondrial morphology. These UDC positive effects may contribute to the observed decrease in fat accumulation and may delay the progression of alcoholic injury to more advanced stages. PMID- 9870713 TI - Iron deposits in the central nervous system of SJL mice with experimental allergic encephalomyelitis. AB - Iron has been proposed to promote oxidative tissue damage in multiple sclerosis (MS). In order to gain insights about how iron gets processed during MS, the deposition of iron was investigated in the CNS of mice with experimental allergic encephalomyelitis (EAE), which is a commonly used animal model of MS. Control mice (adjuvant only) and EAE mice (myelin basic protein plus adjuvant), were sacrificed at 4-8 days (preclinical phase), 10-13 days (clinical phase), or 18 days (recovery phase) post injection. Sections from the cerebrum, hindbrain, and cervical, thoracic and lumbar spinal cord were stained as previously described (J. Neurosci. Res. 29:413, 1991), and scored blindly for histopathological staining. There was minimal histopathological staining at any age in control animals or during the preclinical stage in EAE animals. At the clinical stage of EAE, stained pathological features (macrophages, extravasated RBC and granular staining) were significantly increased compared to the preclinical stage. In the recovery phase, macrophage and granular staining persisted but there was loss of extravasated RBC. Dual labeling studies revealed that granular deposits were present in astrocytes and in locations that appeared to be extracellular. In order to gain insights about the origin of iron deposits in EAE mice, additional studies were performed on brains of mice with extravasated blood lesions. These brains had granular, macrophage and RBC staining. Thus, each of the stained features in EAE animals could be due to the extravasation of blood which occurs in the SJL model of EAE, although some of the iron could have originated from myelin and oligodendrocytes damaged during EAE. PMID- 9870714 TI - Hypoxic and hyperoxic effect on blood phosphodiesterase activity in young and old rats. AB - In the present work we describe phosphodiesterase (PDEs) activity in the blood of young and old rats kept under hypoxic or hyperoxic normobaric conditions in order to correlate the age-change response with PDE levels and oxygen supply. PDE is important in the process of energy supply and as a modulator and mediator of several cellular functions. Three groups of Wistar rats were kept in room air, 10 12% oxygen for 12 days and 98-100% oxygen for 60 hrs respectively. Each group was composed of young rats (2 months of age) and old rats (25 months of age). After the exposure the rats were anaesthetized and blood samples were collected using an intracardiac catheter. The results show: a) in the control group, no significant difference between the PDE activities of old and young rats; b) a significant increase in PDE occurred after hypoxic and hyperoxic treatment in both young and aged rats; c) the increase in PDE activity was more evident in the young rather than the old rats; and d) the aged rats are less responsive to oxygen variation. The results demonstrate that young and aged rats respond to variations in the oxygen supply. Hypoxia and hyperoxia show different age-related intensity level response. We conclude that the alteration in PDE expression occurring in the blood as a consequence of hypoxic or hyperoxic treatment is probably a necessary protective response for the body against alteration from oxidative metabolism and to maintain the body in homeostatic ranges for energy requirements. PMID- 9870715 TI - In vivo imaging of brain nicotinic acetylcholine receptors with 5-[123I]iodo-A 85380 using single photon emission computed tomography. AB - The distribution and kinetics of 5-[123I]iodo-A-85380, a novel ligand for brain nicotinic acetylcholine receptors (nAChRs), were evaluated in the Rhesus monkey using single photon emission computed tomography (SPECT). Peak levels of radioactivity were measured in brain at 90 min after injection of the tracer. Accumulation of radioactivity was highest in the thalamus, intermediate in the frontal cortex and basal ganglia, and lowest in the cerebellum. The ratio of specific to nonspecific binding (V3") in the thalamus, estimated from the (thalamic-cerebellar)/cerebellar radioactivity ratio, reached a value of 6 at 4 h post-injection. Specific binding was reduced by subcutaneous injection of 1 mg/kg cytisine at 2.25 h after injection of radiotracer. At 2.5 h after cytisine administration, radioactivity in the thalamus was reduced by 84%, in the frontal cortex, by 76%, and in the basal ganglia, by 57% of the level measured at the time of cytisine administration, demonstrating that the binding was reversible. On the basis of these findings, together with other data indicating high affinity, receptor subtype selectivity, low nonspecific binding and lack of toxicity in animals, 5-[123I]iodo-A-85380 appears to be a promising ligand for SPECT imaging of nAChRs in the human brain. PMID- 9870716 TI - Calphostin C induces expression of amphiregulin mRNA via reactive oxygen species in IEC-6 cells. AB - Calphostin C, a secondary metabolite of the fungus Cladosporium cladosporioides, is generally used as a specific inhibitor of protein kinase C. It is known that 12-O-tetradecanoyl-13-phorbol acetate (TPA), a protein kinase C activator, induces expression of mRNA for amphiregulin (AR), a member of EGF-related polypeptides, in mammalian epithelial cells. In this work, we determined the effect of calphostin C on AR mRNA expression in IEC-6 cells, a rat intestinal epithelial cell line, and unexpectedly found that this compound enhanced the TPA induced expression of AR mRNA. Moreover, calphostin C alone induced expression of AR mRNA in a light-dependent manner, and this effect was abrogated by pretreatment with N-acetylcysteine. These results suggest that calphostin C can upregulate expression of AR mRNA via reactive oxygen species. PMID- 9870717 TI - Reduction of thyroid hormone may participate in the modulation of cytochromes P450 2C11 and 3A2 by retinol. AB - Both hypothyroidism and retinol supplementation in rats induce CYP 3A2 and suppress CYP 2C11. Therefore studies were performed to evaluate the role of thyroid hormones in the modulation of P450 expression by retinol. Adult male Sprague-Dawley rats were given retinol as a single oral dose of 75 mg/kg. Rats were killed and hepatic microsomes prepared at 24, 48, 72, and 96 hr following treatment. The catalytic activity of 2C11 was reduced maximally by retinol at 48 hr (by 30%) whereas 3A2 activity was elevated maximally at 24 hr (by 30%). The serum concentration of testosterone was not altered at any time point. However, retinol produced a decline in the concentration of thyroxine by 35% and 43% at 24 and 48 hr, respectively. These data suggest that administration of large doses of retinol may alter hepatic microsomal enzyme expression by perturbation of plasma thyroid hormone levels. PMID- 9870718 TI - The British Association of Surgical Oncology, 1973-1998. PMID- 9870719 TI - The British Association of Surgical Oncology Guidelines for surgeons in the management of symptomatic breast disease in the UK (1998 revision). BASO Breast Specialty Group. PMID- 9870720 TI - Screening for colorectal cancer: present, past and future. AB - Colorectal cancer results in 18,000 deaths annually in England and Wales, with 24,000 new cases diagnosed each year. Despite a better understanding of the genetics, and advancement in surgical and anaesthetic techniques, there has been little reduction in mortality and morbidity from this disease over the past 25 years. Colorectal cancer fits recognized criteria for a disease that should be screened in asymptomatic individuals. The putative duration of the adenoma to carcinoma sequence gives an ample window of opportunity to detect and treat colorectal cancer. In this article we have reviewed the strategies involved in screening for colorectal cancer in an asymptomatic population. We have presented trials and arguments for and against the different screening methods and discussed cost effectiveness of screening. In the USA and Canada, major professional organizations and societies now endorse screening; in the UK it is still far from being accepted. We feel that the available evidence shows that colorectal cancer screening has the potential to reduce the morbidity and mortality from this disease and that funding for a mass screening and public education programme should be sought. PMID- 9870721 TI - Gall bladder cancer: proposal for a modification of the TNM classification. AB - Gall bladder cancer is the commonest biliary tract malignancy. The TNM classification of AJCC-UICC is the most widely accepted and most commonly used system for staging. We propose some modifications in the existing classification and recommend guidelines for management based on the stage of the disease. PMID- 9870722 TI - An audit of patient acceptance of one-stop diagnosis for symptomatic breast disease. AB - AIMS: The impetus for optimizing outpatient provision of breast-care services has come both from the patient and management in order to reduce anxiety and make full use of scarce resources. The one-stop diagnostic clinic for the investigation of symptomatic breast lesions is a relatively recent concept with well-known service benefits. However, acceptance to the patient has not been previously investigated. RESULTS: The results of this prospective audit demonstrate a high level of patient satisfaction with the multi-disciplinary, one stop breast clinic. PMID- 9870723 TI - Interactive multimedia information program for use by breast-care nurses--a patient acceptability study. AB - AIMS: To design an interactive multimedia program for use by breast-care nurses and evaluate the acceptability of this technology to patients in the clinical setting. METHODS: In order to ensure that the clarity of the information was maintained the multimedia program was developed by a multidisciplinary team, including non-medical personnel and patients. A prospective analysis of the subjective impressions of patients with symptomatic breast disorders and breast care nurses to a multimedia patient information system was then performed using a standard questionnaire and semi-structured interviews. RESULTS: Fifty women were recruited for the study. Thirty-six (72%) considered the multimedia counselling to be superior to the traditional modalities. Forty-nine (98%) graded the system as good or better. No patient regarded the technology as anxiety-provoking or inferior to the traditional leaflet-based approach. Women over 55 years old found the system as acceptable and easy to use as the younger women. CONCLUSIONS: The multimedia breast counselling programme was acceptable to patients and was considered superior to the traditional leaflet-based approach by the majority. The inherent advantages of this technology will lead to its increasing utilization in the clinical setting. PMID- 9870724 TI - The importance of surgery and accurate axillary staging for survival in breast cancer. AB - AIMS: The purpose of this study was to investigate, within the context of the Danish Breast Cancer Cooperative Group (DBCG) programmes, whether a dedicated surgical approach had a significant bearing on the outcome of breast cancer treatment. METHODS: From 1 January 1980 to 31 December 1990, patients below 70 years of age with operable breast cancer from Odense University Hospital (n=743) were compared with those from the rest of Denmark (denoted rest-DK) (n=15,419). All patients were treated according to nationwide DBCG guidelines and reported to the DBCG Data Centre. The potential median observation time was 11.2 years (range 6.0-16.9). Patients underwent mastectomy or breast conserving therapy, and high risk lymph-node positive patients had adjuvant systemic therapy with or without radiotherapy. RESULTS: Comparing total patients series, overall survival (OS) was significantly superior in patients from Odense compared with rest-DK (P=0.02), with 10-year OSs of 62% (95% CI: 58-65%) and 56% (55-57%), respectively. In subgroups, the OS of low-risk node negative patients (protocol A) in Odense compared with rest-DK was significantly better (P=0.02); 10-year OS was 78% (73 84%) versus 72% (70-73%). Among the high-risk pre-menopausal patients (protocol B), the OS was significantly better in Odense (P=0.009); 10-year OS was 67% (60 75%) versus 53% (51-55%) in rest-DK. Post-menopausal high-risk patients (protocol C) did not differ significantly in OS between Odense and rest-DK (P=0.61). Locoregional control in the Odense series was superior compared with rest-DK. More lymph nodes were recovered and examined from the axilla in the Odense series than in rest-DK, a median of 10 vs. 6 nodes. In the Odense series, a significantly higher proportion of pre-menopausal patients had positive lymph nodes, predominantly one to three positive nodes, and subsequently a lower proportion of pre-menopausal patients had negative lymph nodes compared with rest DK (P=0.02), indicating a more accurate staging in Odense vs. rest-DK. The survival benefit among the patients from Odense cannot be explained by stage migration alone, but seems to represent a true survival advantage. Overall mortality was significantly lower in the Odense series compared with rest-DK. Whether or not this difference could be explained by lower background mortality in the Odense series or was caused by superior treatment is discussed. CONCLUSIONS: The extent of surgery seems important for locoregional tumour control and accurate axillary lymph-node staging. In combination, these might lead to superior recurrence-free and overall survival, although differences in background mortality cannot be ignored. Surgery, therefore, might represent a risk factor by itself. PMID- 9870725 TI - Conservative treatment of lower pole breast cancers by bilateral mammoplasty and radiotherapy. AB - AIMS: This series analyses the results of conservative surgery for large lower pole breast cancers by lumpectomy associated with a bilateral remodelling mammoplasty, in order to avoid residual deformities. METHODS: This retrospective study concerns 50 patients with a lower pole breast cancer treated between 1986 and 1996 by lumpectomy, mammoplasty and irradiation. The contralateral breast was immediately made symmetrical in all cases. The mean tumour size was 32.5 mm. RESULTS: The mean weight of the lumpectomy specimen was 270 g. Resection margins were tumour-free in 90% of cases. The main complication observed was delayed healing, thus postponing post-operative treatment in 6.5% of cases. The median follow-up was 48 months. The 5-year actuarial ipsilateral local recurrence rate was 7% and 5-year actuarial metastasis-free and overall survival rates were 81 and 97%, respectively. Cosmesis was satisfactory in 85% of patients. We observed better results when radiotherapy was performed after rather than prior to surgery (92 vs. 67%: NS). CONCLUSIONS: Performing a bilateral mammoplasty at the time of initial surgery for large breast cancers situated in the lower quadrants of the breast facilitates larger lumpectomies with good cosmetic results. PMID- 9870726 TI - Gamma probe-guided sentinel node biopsy--optimal timing for injection. AB - AIMS: We initiated a Phase I feasibility study using a gamma-detecting probe (GDP) and radiolabelled colloid to localize the sentinel lymph node (SLN) in breast cancer. The aim of the study was to establish the ideal timing for injection and examine any possible exclusion criteria for this method. METHODS: Thirty breast cancer patients diagnosed by fine needle aspiration (FNA) were included in this study. All were injected with 60 MBq rhenium colloid labelled with 99mTc (Tck-17). Scintigraphy was done 20 min, 2, 6 and 25 hours post injection. Patients were then taken to surgery where they were injected with patent blue dye. During surgery, the SLN was located with a GDP (Neoprobe Model 1000). In 28 patients, the SLN was identified by scintigraphy 2 hours after injection, identical to the images seen after 24 hours. RESULTS: In all 28 patients, the SLN was found by the GDP during surgery. In 26 patients the SLN was dyed blue. The two patients with no SLN localization had received prior radiation. Pathology disclosed SLNs with metastases in seven patients. Two patients had a negative SLN but had an axillary lymph node replaced by tumour. CONCLUSIONS: Two to 24 hours prior to surgery is suitable timing for injection. Previous radiotherapy predicts failure for this procedure. Further studies are needed to find the exact false-negative rate of this method for breast cancer. PMID- 9870727 TI - Technique of lymphatic mapping and sentinel node biopsy for melanoma. AB - AIMS: An increasing number of surgeons perform sentinel node biopsy to identify melanoma patients with early lymphatic dissemination who may benefit from regional node dissection or adjuvant therapy. The addition of lymphoscintigraphy and intraoperative gamma-ray detection with a hand-held probe increases the sensitivity of the surgical technique substantially. METHODS: The value of lymphoscintigraphy is discussed. The operative technique of lymphatic mapping and sentinel node biopsy is described, including the use of a vital dye and a gamma ray probe. CONCLUSIONS: Close to 100% of first-tier lymph nodes can be identified with this combined approach without the unnecessary removal of too many higher echelon nodes. PMID- 9870728 TI - The incidence and Dukes' staging of colorectal cancer over 3 decades. AB - AIMS: Two previous studies (1966-1971 and 1979-1983) of patients with colorectal cancer (CRC) have been reported from our hospital. A large increase in the incidence of CRC was noted, and an improvement in Dukes' staging of tumours at treatment. We report a series of patients admitted with newly diagnosed CRC to evaluate this trend further. METHODS: A prospective study was made of all patients with newly diagnosed CRC admitted to the John Radcliffe Hospital, Oxford in 1995. Means of diagnosis and Dukes' staging were recorded. RESULTS: In 1 year 177 patients were admitted with newly diagnosed CRC. Previous studies had shown an increase from 52.8 to 103.4 patients per year. The number of patients diagnosed by colonoscopy doubled from 19.4% in 1979-1983 to 41% in 1995. No significant change in the proportion of patients with Dukes' A or B tumours was found. CONCLUSION: The number of patients treated annually with CRC in a stable population has more than trebled in the last 3 decades. A real increase in the incidence of CRC is likely as this rise cannot be explained solely by changing referral patterns or an ageing community. There are no significant changes in presentation patterns despite the availability of colonoscopy since 1975. PMID- 9870729 TI - Local recurrence in patients with rectal cancer diagnosed between 1988 and 1992: a population-based study in the west Netherlands. AB - AIMS: We carried out a population-based study of local recurrence rates in curatively resected patients with rectal cancer, diagnosed between 1988 and 1992. The first objective was to make an inventory of the overall local recurrence rate after non-standardized conventional surgery, inter-institutional recurrence rate variability, and correlations between patient- and tumour-related factors and recurrence rate. A second objective was to investigate the compliance to guidelines for post-operative radiotherapy. METHODS: Data were obtained from the Comprehensive Cancer Centre West. The study comprised 1105 patients from 12 hospitals. Of these patients, 437 were ineligible because of missing medical records, no carcinoma, incorrect registration, no laparotomy, non-curative resection, or loss to follow-up. RESULTS: The overall local recurrence rate was 22.5% with a range of 9-36% between the hospitals. These differences were not significant. Dukes' Astler-Coller stage, tumour location, and residual tumour were significant independent prognostic factors for the risk of local recurrence. Indications for post-operative radiotherapy were Dukes' Astler-Coller B2 and C tumours, positive surgical margins, and tumour spill. Compliance to the guidelines for radiotherapy was only 50%. However, no significant difference in recurrence rate was found between patients treated according to the guidelines and those not treated according to the guidelines. CONCLUSION: This study shows a large variability in local recurrence rate between the participating hospitals and confirms that the risk of recurrence in primary rectal cancer is dependent on Dukes' Astler-Coller stage, tumour location and residual tumour. Furthermore, this study contributes to the discussion about the feasibility of guidelines for post-operative radiotherapy. PMID- 9870730 TI - Improvements in staging of gastric carcinoma from using the new edition of TNM classification. AB - AIMS: To recognize possible advantages of the 5th edition of TNM of gastric carcinoma in comparison to the former edition. METHODS: Data from Erlangen Cancer Center for 898 patients with gastric carcinoma treated surgically by total or subtotal gastrectomy with en bloc lymphadenectomy were analysed. RESULTS: The prognostic significance of TNM has been improved by the new edition as demonstrated by the likelihood ratio test. In addition, uncertainties of method in the pathological classification of regional lymph-node metastasis inherent in the 4th edition can now be avoided. CONCLUSION: The changes in the TNM classification introduced by the 5th edition are justified by data from the Erlangen Cancer Center, have methodic advantages in determining the N classification, and lead to an improvement in estimation of outcome. PMID- 9870732 TI - Male breast cancer: a 22-year experience. AB - AIMS: To carry out a retrospective study of male breast cancer over a 22-year experience. METHODS: Data from 121 male patients with breast cancer treated between the years 1972 and 1994 at the Surgical Clinic of Ankara Oncology Hospital were reviewed. Distribution of cases according to stage was: 2.5% stage I, 28.9% stage II, 55.4% stage III and 13.2% stage IV (AJCC staging method). The surgical treatment for 23 of the patients (19%) was Halsted's radical mastectomy or modified radical mastectomy. Seventy-three cases (60.3%) had total mastectomy without axillary node dissection and 25 (20.7%) had local tumour excision only. Seventy-two of 121 patients had adjuvant treatment. RESULTS: In general the prognosis of men with breast cancer was worse than for women. In the analysis of patients in stages I, II and III-A (operable disease group), the 5-year survival rates were 73% in axillary node-negative patients and 77% in those with tumours sized under 5 cm (P<0.001). In these patients, univariate analysis demonstrated that axillary status (relative risk of death in positive status vs. negative=3.6), tumour size (relative risk in T3 vs. T1-2=2), surgical treatment type (relative risk in simple mastectomy vs. radical mastectomy=1.9) and adjuvant chemotherapy (relative risk if no chemotherapy=1.4) were statistically significant factors associated with survival. CONCLUSIONS: Cox's regression model revealed that axillary status, tumour size and type of surgical treatment were the most important independent prognostic factors (P<0.001). PMID- 9870731 TI - Isolated hypoxic perfusion with mitomycin C in patients with advanced pancreatic cancer. AB - AIMS: Chemotherapy as a palliative therapy option for patients with advanced pancreatic cancer remains disappointing. Some authors have recently claimed high response rates and a prolongation of median survival after regional chemotherapy. Isolated perfusion may result in the highest drug concentrations within the target tissue without causing systemic side-effects. An established, commercially available system of isolated hypoxic perfusion (IHP) was therefore evaluated in patients with unresectable or recurrent pancreatic cancer. PATIENTS AND METHODS: IHP was performed in 17 patients with unresectable pancreatic cancer. Five women and 12 men with a median age of 61 years were treated. A 20-min isolated hypoxic perfusion was performed after 40 mg of mitomycin C (MMC) had been instilled into the running perfusion system over 5 min. Tumour response was evaluated by CT-scan 6 weeks after IHP. RESULTS: Twenty perfusions were carried out in 17 patients. Within 10 min of perfusion, the perfusate's PO2 decreased to 13% of the baseline value. Five minutes after the infusion of MMC a local concentration of 15.2 mg/litre was observed. Toxicity-related deaths did not occur. Nausea and vomiting (NCI> or =II: 12 episodes) were the most frequently observed toxicities after IHP. In five patients (29%) a deep vein thrombosis occurred. None of the treated patients responded to the regimen used in this trial. The median survival time after IHP was 4.2 months (range 1.3-21). CONCLUSIONS: The overall rate of side effects and complications after IHP was high. In spite of some hopeful reports on this treatment in recent years, IHP did not show any benefit in terms of tumour response or median survival. On the basis of these experiences, this procedure should no longer be used as treatment for patients with unresectable or recurrent pancreatic cancer. PMID- 9870733 TI - Role of serum thyroglobulin in the pre-operative evaluation of follicular thyroid tumours. AB - AIMS: There is no reliable diagnostic test for pre-operative differentiation between benign and malignant follicular and Hurthle cell neoplasms. Measurements of serum thyroglobulin (Tg) are currently only used post-operatively as a marker of recurrent disease or distant metastases in the follow-up of patient with differentiated thyroid cancer. In this study pre-operative serum Tg measurements were performed with the aim of investigating whether Tg levels differ in benign and malignant follicular and Hurthle cell neoplasms. METHODS: In 516 patients who underwent thyroid surgery at the Institute of Oncology in Ljubljana, Slovenia, from 1990 to 1996, serum Tg concentration was measured in addition to the standard pre-operative tests (fine-needle aspiration biopsy, ultrasonography, 99mTc scanning and hormonal profile). After the operation, patients were divided into 11 groups based on their histological diagnosis (papillary cancer--classic, papillary cancer--follicular variant, papillary cancer oncocytic variant, occult papillary cancer, follicular adenoma, follicular cancer, Hurthle cell adenoma, Hurthle cell cancer, anaplastic cancer, medullary cancer, nodular goiter) and the serum Tg values of the different groups were compared. RESULTS: In groups of patients with follicular and Hurthle cell cancer, median Tg values were higher (2895 and 638.5 ng/ml) and, statistically, differed significantly from the serum Tg values in all other groups (P<0.01). Sensitivities and specificities of the tests were 71.8% and 80.4% for follicular cancer and 55.6% and 83.8% for Hurthle cell cancer, while positive and negative predictive values were 75.6% and 77.1% for follicular cancer and 75% and 68.4% for Hurthle cell cancer. CONCLUSIONS: These results indicate that pre-operative serum Tg measurements might be an important additional diagnostic tool in the pre-operative work-up of patients with thyroid tumours. PMID- 9870734 TI - Lipiodol avidity of neuroendocrine liver metastases. AB - AIMS: Lipiodol has been shown to concentrate in most hepatocellular carcinomas as well as in some liver metastases, including those of neuroendocrine origin. Our aim was to determine the proportion of neuroendocrine liver metastases that take up lipiodol and to identify tumour characteristics that predict avidity. METHODS: Avidity was assessed in 12 patients with neuroendocrine liver metastases by performing an abdominal CT scan immediately after selective hepatic arterial injection of 5 ml of unlabelled lipiodol and this was correlated with number and size of lesions as well as angiographic and plain CT scan features. RESULTS: In seven patients the tumours displayed lipiodol avidity (four solitary, three multiple); five patients had non-avid lesions (all multiple). A large dominant liver tumour was the only predictor of avidity (mean diameter of largest lesion 9 cm vs. 3 cm for patients with non-avid tumours: P=0.01). Avidity was not related to vascularity or CT density of lesions. CONCLUSIONS: Although this is a small study, it would appear that approximately 50% of neuroendocrine liver metastases selectively concentrate lipiodol, which could have implications for targeted cancer therapy. PMID- 9870736 TI - The evolutionary biology of death and human malignancy. PMID- 9870735 TI - Necrolytic migratory erythema, first symptom of a malignant glucagonoma: treatment by long-acting somatostatin and surgical resection. Report of three cases. AB - We report three cases of malignant glucagonoma with necrolytic migratory erythema as the first clinical symptom. Long-acting somatostatin analogue was the first step of a multimodal therapeutic strategy which included surgical resection of the primary tumour in every case. Liver metastases which were present in two patients were treated by hepatic arterial chemoembolization and systemic chemotherapy in one case and by liver resection for cytoreduction and hepatic arterial chemoembolization in another case. Skin lesions resolved in all three patients. PMID- 9870737 TI - Breast cancer imaging--what are the optimal modalities? PMID- 9870738 TI - Historical, current and future aspects of osteosarcoma treatment. PMID- 9870739 TI - Carcinoid somatostatinoma of the duodenum. AB - Carcinoid somatostatinoma is a rare neuroendocrine malignant tumour and the duodenal location is an atypical site of presentation of which only few cases have been reported in the literature. A case of duodenal carcinoid somatostatinoma metastatic to lymph nodes in a 66-year-old patient is presented with an update of the literature. No relevant signs or symptoms were associated to the retrogastric lymph-node mass, which deformed but did not infiltrate the stomach wall. At the first and third portion of the duodenum, two polipoid endoluminal nodules (size 1 cm) were found with adjacent adenopathy partially adherent to the head of the pancreas and with thickening of the antropyloric wall. The patient underwent antrectomy duodenum mobilization and lymphadenectomy in the hepatic artery region. The treatment was successful and, over 3 years after diagnosis, there has been no clinical or radiological evidence of relapse. Duodenal somatostatinoma is rare and its diagnosis is often incidental. Surgery would be the appropriate treatment in the early stage of the disease with good chances of cure. PMID- 9870740 TI - Carcinoma of the gallbladder presenting as scalp tumour. AB - Carcinoma of the gallbladder is characterized by rapid tumour growth associated with lymphatic and local tumour invasion. The peritoneum, GIT and lungs are common sites of seeding. Distant metastasis to bone rarely occurs. Here we document a case of silent gallbladder carcinoma presenting as scalp tumour with improved survival. PMID- 9870741 TI - Long-term survival of a female patient with primary malignant melanoma of the urethra. AB - Primary malignant melanoma of the female urethra comprises 0.2% of all melanomas and has poor prognosis. In the period 1972-1992, 75 cases of primary urethra carcinomas were treated at the Roswell Park Cancer Institute. Among them, only an 80-year-old woman was diagnosed with primary malignant melanoma. Despite conservative treatment, she lived for 7 years. We believe that local surgical excision can be an option for treatment in selected patients as it retains quality of life. PMID- 9870742 TI - Palisaded myofibroblastoma--an uncommon tumour of lymph nodes. AB - We report a case of palisaded myofibroblastoma of the groin in a woman aged 20 years who presented with a large groin swelling. It is an uncommon benign tumour mainly arising in a lymph node. PMID- 9870743 TI - Transosseous transposition of a pedicled rectus abdominis flap to cover hip wounds. AB - Bone-marrow transplantation may be complicated by aseptic necrosis of the femoral head. Prolonged immunosuppression places the patient at increased risk of infection and is of particular concern if replacement arthroplasty is contemplated. Infection of a prosthesis usually requires thorough debridement and removal of the device. However, attention must also be paid to the resultant cavity and soft-tissue cover. Several flaps have been reported for this problem, though limitations of flap size may make it difficult to achieve both objectives. This report describes the use a pedicled rectus abdominis flap that was tunnelled through the acetabulum in order to gain direct access to the hip and allow adequate length both to fill the dead-space and to provide cover. PMID- 9870744 TI - Primary angiosarcoma of bone in Paget's disease. AB - We report here a case of a primitive angiosarcoma of the bone occurring in Paget's disease. Only one case has been previously described in the literature. We review other types of tumours occurring in Pagetic bone and discuss medical and surgical management. PMID- 9870745 TI - Dendritic release of vasopressin and oxytocin. AB - In addition to the release of neurotransmitters from their axon terminals, several neuronal populations are able to release their products from their dendrites. The cell bodies and dendrites of vasopressin- and oxytocin-producing neurones are mainly located within the hypothalamic supraoptic and paraventricular nuclei and neuropeptide release within the magnocellular nuclei has been shown in vitro and in vivo. Local release is induced by a range of physiological and pharmacological stimuli, and is regulated by a number of brain areas; locally released peptides are mainly involved in pre- and postsynaptic modulation of the electrical activity of magnocellular neurones. Spatial and temporal differences between peptide release within the nuclei and that from the distant axonal varicosities indicate that the release mechanisms are at least partially independent, supporting the hypothesis of locally regulated dendritic release of vasopressin and oxytocin. In this respect, magnocellular neurones show similarities to other neuronal populations and thus autoregulation of neuronal activity by dendritic neuromodulator release may be a general phenomenon within the brain. PMID- 9870746 TI - Role of non-NMDA receptors in osmotic and glutamate stimulation of vasopressin release: effect of rapid receptor desensitization. AB - Previous studies demonstrated that the increase in vasopressin (VP) release and induction of VPmRNA content by osmotic stimulation was blocked by kynurenic acid, a non-specific antagonist of excitatory amino acid (EAA) receptors. In order to identify the type of EAA receptor involved, perifused explants of the hypothalamo neurohypophyseal system (HNS) were exposed to a ramp increase in osmolality (40 mOsm over 6 h achieved by increasing NaCl) in the presence and absence of 10 microM 6,7-dinitroquinoxaline-2,3-dione (DNQX), an antagonist of non-n-methyl-d aspartate (NMDA) excitatory amino acid receptors. Vasopressin release and VP mRNA content were significantly increased by exposure to the osmotic stimulus. 6,7 dinitroquinoxaline-2,3-dione inhibited osmotically stimulated VP release (F=16.65, P=0.0008) without significantly reducing basal release. It also prevented the osmotically stimulated increase in VP mRNA content (P <0.05). Although these results implicated glutamate, the primary endogenous ligand for EAA receptors, in the regulation of VP, exogenous glutamate was ineffective in stimulating VP release from HNS explants in either low-Mg2+ or Mg2+-replete medium. However, blockade of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor desensitization with cyclothiazide (100 microM) caused a marked increase in VP release in response to 100 microM glutamate, and blockade of kainate receptor desensitization with concanavalin A resulted in a small, but significant increase in VP release in response to 1 mM glutamate. These results support a role for non-NMDA receptor activation in osmotic regulation of VP release. PMID- 9870747 TI - Role of adrenoceptors in vasopressin, oxytocin and prolactin responses to conditioned fear stimuli in the rat. AB - Conditioned fear or novel environmental stimuli suppress vasopressin (VP) and augment oxytocin (OT) and prolactin (PRL) release in rats. We examined the effects of intracerebroventricular (i.c.v.) injections of adrenoceptor antagonists on these neuroendocrine responses to conditioned fear or novel environmental stimuli in male rats. A beta1 antagonist, metoprolol, blocked the VP but not the OT or PRL response to conditioned fear stimuli, but did not abolish neuroendocrine responses to novel environmental stimuli. A beta2 antagonist, ICI118551, impaired the PRL but not the VP or OT response to fear or novel environmental stimuli. In rats injected with a alpha1 adrenoceptor antagonist, benoxathian, conditioned fear stimuli did not significantly induce the VP, OT or PRL responses. The effects of benoxathian were not due to a general reduction of arousal, since benoxathian did not prevent the VP, OT or PRL response to novel environmental stimuli. These data suggest that beta1 adrenoceptors play a selective role in the VP response to conditioned fear stimuli, as do beta2 adrenoceptors in the prolactin response to conditioned fear and novel environmental stimuli. We conclude that alpha1 adrenoceptors play a facilitative role in VP, OT, PRL responses to conditioned fear stimuli. PMID- 9870748 TI - Noradrenergic neurotoxin suppresses gonadotropin-releasing hormone (GnRH) and GnRH receptor gene expression in ovariectomized and steroid-treated rats. AB - The present study was designed to investigate whether noradrenergic neurotransmission regulates the gene expression of gonadotropin-releasing hormone (GnRH) in the preoptic area and GnRH receptor in the pituitary. To this end, N-(2 chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4, 50 mg/kg), an intraperitoneal (i.p.) injection of selective noradrenergic neurotoxin, was administered 1 h before progesterone (1 mg) treatment in ovariectomized and estradiol-treated prepubertal rats. Treatment with DSP4 effectively blocked the progesterone induced increase in hypothalamic noradrenaline content, but not dopamine content, indicating that DSP4 selectively inhibits noradrenergic neurotransmission. DSP4 significantly blocked progesterone-induced increase in serum luteinizing hormone (LH) concentrations as well as GnRH release from hypothalamic fragments incubated in vitro. DSP4 concomitantly down-regulated GnRH mRNA levels in the preoptic area, as determined by competitive reverse transcription-polymerase chain reaction. DSP4 also clearly down-regulated progesterone-induced GnRH receptor mRNA levels in the pituitary, whereas it failed to alter LHbeta mRNA levels. In summary, blockade of noradrenergic neurotransmission with DSP4 resulted in profound reductions of hypothalamic GnRH and pituitary GnRH receptor gene expression. PMID- 9870749 TI - The effects of oestrogen and progesterone on serotonin and its metabolite in the lateral septum, medial preoptic area and ventromedial hypothalamic nucleus of female rats. AB - The effects of sex steroid hormones on serotonin and its metabolite, 5 hydroxyindole-3-acetic acid (5-HIAA) in the lateral septal nucleus (LS), the medial preoptic area (MPA) and the ventromedial nucleus of the hypothalamus (VMH) of female rats were investigated, using immunohistochemistry and high-performance liquid chromatography (HPLC). Female rats were divided into three groups: ovariectomized rats (OVX group); OVX-rats treated with estradiol benzoate alone (E2 group); and OVX-rats treated with E2 plus progesterone (E2 + P group). We analysed the density of serotonin-immunoreactive fibres with a computer-assisted image analysis system, and measured the tissue concentrations of serotonin and 5 HIAA. Many serotonin-immunoreactive fibres were observed in the LS, MPA and VMH in all three groups. The density of serotonin-immunoreactive fibres in the MPA and VMH was significantly lower in the E2 and E2+P groups compared to the OVX group, whereas the LS showed no detectable differences among the three groups. In the HPLC study, the concentrations of serotonin in the MPA and VMH of the E2 and E2+P groups were significantly lower than that in the OVX group. There was no significant difference in the concentration of serotonin in the LS. The concentration of 5-HIAA and the ratio of 5-HIAA/serotonin in the LS, MPA and VMH showed no significant differences among the OVX, E2 and E2+P groups. The present results suggest that E2 priming for sexual behaviour can affect the serotonergic system by decreasing serotonin content, but not the turnover rate, in the MPA and VMH of female rats. PMID- 9870750 TI - Actions of prostaglandin E2 on rat supraoptic neurones. AB - Prostaglandins (PGs) have been implicated in the regulation of vasopressin (VP) and oxytocin (OT) release in response to various stimuli. To examine the site and mechanism of actions of PGs, we studied effects of PGE2 and PG-receptor agonists on supraoptic nucleus (SON) neurones of rat hypothalamic slice preparations using extracellular recording and whole-cell patch-clamp techniques. PGE2 modulated the electrical activity of more than 80% of the neurones studied. The effects of PGE2 on both phasic and non-phasic neurones were mostly excitatory, and dose dependent. The effects of PGE2 were mimicked by PGF2alpha or the FP agonist, fluprostenol, whereas PGD2 or the selective EP, IP or TP agonist was less effective or had no effect. The effects of PGE2 were unaffected by the EP1 antagonist, SC-51322, but reduced to 80% of control by the EP1/FP/TP antagonist, ONO-NT-012, which reduced the effects of fluprostenol to 32% of control. Moreover, some neurones responsive to PGE2 did not respond to fluprostenol. Patch clamp analysis in SON slice preparations revealed that PGE2 at 10(-6) M depolarized the membrane potential by 3.9+/-0.3 mV from the resting membrane potential of -58.4+/-2.2 mV in the current-clamp mode. In the voltage-clamp mode, PGE2 induced inward currents at a holding potential of -70 or -80 mV, while it did not affect spontaneous excitatory postsynaptic currents. PGE2 induced currents also in dissociated SON neurones and the reversal potential of the currents was -35.5+/-0.9 mV, which was similar to that of currents induced by fluprostenol. These results suggest that SON neurones possess at least two types of PG receptors, FP receptors and EP receptors of a subclass different from EP1, EP2, or EP3, and that activation of these receptors leads to the opening of nonselective cation channels, membrane depolarization and increase of the action potential discharge. PMID- 9870751 TI - Diurnal changes in the expression of genes encoding for arginine vasotocin and pituitary pro-opiomelanocortin in the rainbow trout (Oncorhynchus mykiss): correlation with changes in plasma hormones. AB - Using quantitative in-situ hybridization, this study monitored diurnal changes in the abundance of the gene transcripts of two corticotropin-releasing peptides, arginine vasotocin (AVT) and isotocin in hypothalamic neurones, and of pro opiomelanocortin (POMC) mRNA in the pituitary of the rainbow trout (Oncorhynchus mykiss). A significant diurnal pattern of gene expression was only displayed in the hypothalamus by the parvocellular AVT neurones of the preoptic nucleus. Abundance of AVT mRNA in these neurones was low at lights on (06.00 h), increased during the morning to reach a plateau of peak values between 14.00 h and 22.00 h, and then declined during the dark phase. This pattern was the inverse of that shown by plasma cortisol values. Changes in AVT transcript abundance are also considered in terms of the reported diurnal change in circulating AVT concentration. Pituitary and hypothalamic AVT peptide content did not change. Transcripts of both POMC genes (POMC-A and POMC-B) were monitored in pituitary corticotropes and melanotropes. Only POMC-A mRNA was detected in corticotropes where it showed no diurnal change in abundance. Transcripts of both POMC genes were found in the melanotropes, although, judging from autoradiographic intensity, POMC-A mRNA predominated. Both genes showed diurnal differences in their transcription with POMC-A mRNA showing peak values at 10.00 h and a nadir at 02.00 h, while POMC-B mRNA showed an inverse pattern. The results indicate that the two POMC genes can be independently regulated. PMID- 9870752 TI - Relationship between composite matrix molecular structure and properties. AB - The synthesis of three novel dimethacrylate esters for use as the monomer phase in dental composites was studied. The monomers were prepared by the reaction of glycidyl methacrylate with phthalic, isophthalic and terephthalic acids, respectively, and the reaction products were found to be mixtures of isomers. The monomers obtained from the reaction of phthalic and isophthalic acids were low viscosity liquids and the relationship between viscosity and molecular structure was studied. The low viscosity of these di-hydroxyl containing monomers was shown to be due to intramolecular hydrogen bonding. This reduced monomer viscosity compared with BisGMA allowed a reduction in the quantity of diluent monomer triethyleneglycoldimethacrylate (TEGDMA) required to give standard viscosities of 1 and 2 Pas resulting in a modest decrease (ca. 10%) in polymerization shrinkage compared with that exhibited by BisGMA/TEGDMA solutions of equivalent viscosities. A number of properties relevant to the use of these as potential core monomers in dental composites were determined for both monomer and polymer systems. PMID- 9870753 TI - Plasma sprayed hydroxyapatite coatings on titanium substrates. Part 1: Mechanical properties and residual stress levels. AB - Hydroxyapatite (HA) coatings have been sprayed on to substrates of Ti-6Al-4V, using a range of input power levels and plasma gas mixtures. Coatings have also been produced on substrates of mild steel and tungsten, in order to explore certain aspects of the mechanical behaviour of HA without the complication of yielding or creep in the substrate. Studies have been made of the phase constitution, porosity, degree of crystallinity, OH ion content, microstructure and surface roughness of the HA coatings. The Young's moduli in tension and in compression were evaluated by the cantilever beam bend test using a tungsten/HA composite beam. The flexural Young's modulus was determined using a free-standing deposit under the same test. Adhesion was characterised using the single-edge notch-bend test; this is considered superior to the tensile bond strength test in common use. Measured interfacial fracture energies were of the order 1-10 J m( 2). Stress levels were investigated using specimen curvature measurements in conjunction with a numerical process model. The quenching stress for HA was measured to be about 10-25 MPa and the residual stress level in HA coatings at room temperature are predicted to lie in the approximate range of 20-40 MPa (tensile). These residual stresses could be reduced in magnitude by maintaining the substrate at a low temperature (possibly below room temperature) during spraying and it may be worthwhile to explore this. Ideally, the HA coating should have low porosity, high cohesive strength, good adhesion to the substrate, a high degree of crystallinity and high chemical purity and phase stability. In practice, such combinations are rather difficult to achieve by just varying the spraying parameters. PMID- 9870754 TI - Plasma sprayed hydroxyapatite coatings on titanium substrates. Part 2: optimisation of coating properties. AB - Heat treatment and the introduction of a Ti bond coat have been applied to hydroxyapatite (HA) coatings sprayed using different plasma powers and gas mixtures. Attempts were made in this way to achieve optimal coating properties for orthopaedic implants. In particular, the effects on the degree of crystallinity, the adhesion, the OH ion content and the purity were evaluated. Heat treatment at 700 C for 1 h in air proved to be effective in increasing the crystallinity, regaining the OH- ion and removing other non-HA compounds, although it caused a significant decrease in the degree of adhesion (interfacial fracture toughness) for those specimens sprayed at high powers. This heat treatment was found to induce significant transformation of amorphous HA to the crystalline form, while not detrimentally changing the properties of the underlying Ti-6Al-4V substrates. Precoating with a 100 microm Ti layer increased the adhesion of the HA coatings on Ti-6Al-4V substrates, primarily by providing a rougher surface and promoting better mechanical interlocking. Changes in coating properties during immersion in biological fluids were also studied and were found to depend critically on the chemical composition of the fluids. Small precipitates formed on the coating surfaces when immersed in Ringers solution. These might account for the apparent drop in the degree of crystallinity when measured using X-ray diffraction. A significant drop in the interfacial adhesion was found for those coatings sprayed at high powers. This could be offset by prior precoating with a titanium bond coat and suitable heat treatment. In summary, the following processing sequence is suggested in order to achieve optimum coating properties: precoating the substrate with a layer of Ti (approximately 100 microm), spraying HA at a sufficiently high-power level (depending on particle size and gas mixture) and heat treatment at 700 degrees C for 1 h in air. PMID- 9870755 TI - Chemical changes during in vivo degradation of poly(anhydride-imide) matrices. AB - The in vivo degradation characteristics of a novel class of biodegradable polymers, poly(anhydride-imides), were investigated. The poly(anhydride-imides) examined were poly[trimellitylimidoglycine-co-1,6-bis(p-carboxyphenoxy)hex ane] (TMA-gly:CPH) in 10:90, 30:70 and 50:50 molar ratios and poly[pyromellitylimidoalanine-co-1,6-bis(p-carboxyphenoxy)he xane] (PMA-ala:CPH) in 10:90 and 30:70 molar ratios. The polymer matrices were compression-molded into circular discs, then implanted in rat subcutaneous tissues for nearly two months. At defined time intervals, the animals were sacrificed and explants analyzed. Proton NMR spectroscopic analysis revealed a complete absence of imide monomer units in PMA-ala: CPH compositions after 28 d and complete removal of imide units at 56 d from TMA-gly matrices. Gross observation of the implants closely correlated to the imide content: with decreasing imide content, the explants darkened and fragmented at a faster rate. The chemical compositions of the poly(anhydride imide) explants were also monitored using IR spectroscopy. The residual amount of anhydride bonds in the polymer backbone following implantation were calculated from peaks specific to the anhydride bonds relative to the total amount of carbonyl bonds present. Initially, the imide (TMA-gly or PMA-ala) anhydride bonds were rapidly hydrolyzed then solubilized, followed by the slower hydrolysis of the CPH monomer anhydride bonds. PMID- 9870756 TI - Accelerated study on lysozyme deposition on poly(HEMA) contact lenses. AB - A technique was developed to accelerate lysozyme deposition on poly(HEMA) contact lenses and measure the amounts of the deposited lysozyme. This technique was for evaluation of bendazac lysine solution, a contact lens cleaning and wetting solution. Effect of temperature on lysozyme deposition on poly(HEMA) contact lenses was examined. Five temperatures ranging from 25 degrees C to 90 degrees C were chosen to examine the temperature effect. The amounts of lysozyme deposited on poly(HEMA) contact lenses at 25 C and 60 C were 0.27 microg/lens and 0.61 microg/lens, respectively. The amount increased sharply to 23 microg/lens at 70 degrees C with the maximum of 31 microg/lens at 90 degrees C. Kinetics of lysozyme deposition on poly(HEMA) contact lenses was examined at 80 degrees C. Lysozyme deposition increased sharply during the first 2 h and reached a plateau after 2 h. Effectiveness of various cleaning procedures was examined using bendazac lysine solution. When the contact lenses were washed without rubbing with fingers, the bendazac lysine reduced the amount of deposited lysozyme by more than 40% from 18.3 microg/lens to 10.6 microg/lens. The effect of bendazac lysine was most prominent when the contact lenses were shaken during storage in the presence of lysozyme in solution. If the contact lenses were cleaned by rubbing with fingers, the effect of bendazac lysine solution on the prevention of lysozyme deposition was negligible. PMID- 9870757 TI - Initial histological evaluation of anti-washout type fast-setting calcium phosphate cement following subcutaneous implantation. AB - Anti-washout-type fast-setting calcium phosphate cement using chitosan (aw FSCPC(chi)), conventional CPC (c-CPC), CPC mixed with citric acid (CPC(citric)) and CPC mixed with polyacrylic acid (CPC(acrylic)) were implanted subcutaneously in rats immediately after mixing to shed some light on the understanding of the appearance of excellent tissue response to CPC. CPC(citric) and CPC(acrylic) set quickly, similar to aw-FSCPC(chi), but the former two stopped their transformation to apatitic minerals. The c-CPC, which required a long setting time, was found to be crumbled, but the other CPCs maintained the shape at implantation. The aw-FSCPC(chi) and CPC(citric) showed no inflammatory response whereas c-CPC and CPC(acrylic) showed an inflammatory response one week after implantation. A component of the aw-FSCPC(chi) and c-CPC was an apatitic mineral whereas CPC(citric) and CPC(acrylic) showed no transformation to apatite. We concluded that the non-crumbling property plays a more dominant role in the appearance of excellent tissue response to CPC than the transformation to apatite. Also, a non-crumbling property is not a sufficient condition, but a necessary condition for the appearance of the excellent tissue response to CPC. PMID- 9870758 TI - Investigation of a rifampin, fusidic-acid and mupirocin releasing silicone catheter. AB - After strict hygienical measures have been exhausted the use of plastic materials with antibacterial activity may reduce catheter related-bacterial colonization. An antimicrobial silicone catheter was investigated by HPLC-measurement, SEM, antimicrobial assays and standard biocompatibility tests. The modified catheter was highly biocompatible and the antimicrobial leaching non-toxic. The initially release rate was governed by the drug solubility in the 'sink' and surface loading ('burst effect'). The second continuous period depended on the drug velocity in the silicone matrix and was extended up to 100 days with a proportionality to square root of t for each drug. Diffusion exponents were in range of 2 x 10(-8) to 1 x 10(-9) (cm2 sec(-1)). The lower diffusion exponent of mupirocin was explained by its higher cohesion energy and lower physico-chemical compatibility with the embedding silicone. The antimicrobial drugs were in a molecular-dispersed state with the silicone-matrix, whereas superficially located crystals of the antibiotics covering the catheter surface could be demonstrated by SEM. PMID- 9870760 TI - Microdiffraction studies of bone tissues using synchrotron radiation. AB - This is the first report on a series of studies of the crystallinity of bone tissues. The measurements were done at the microfocused diffraction beamline at ERSF (European Synchrotron Radiation Facility) as a feasibility test on various aspects on microdiffraction analysis. Beside the crystal structure, crystallite size distribution and preferential orientation were also studied, with a spatial resolution of 7 microm. The experiments were performed at the microfocus beam line 1 at European Synchrotron Radiation Facility (ESRF). The samples were longitudinal and transversal cuts from human femoral shafts. Over 65 diffraction lines from apatite structure could be identified. The results show that the bone crystallites seem to be oriented in two orthogonal directions, one parallel with the Haversian system and the other perpendicular following the lamella's curvature. Peak width analysis shows that the crystallites are about 25-40 nm along the c-axis with a width of about 10 nm. The result also shows that the peak width is somewhat larger close to the haversian canal compared with the outer region of the osteon. PMID- 9870759 TI - In vivo and in vitro responses to poly(ethylene terephthalate-co-diethylene glycol terephthalate) and polyethylene oxide blends. AB - Although biocompatible polymeric compounds are generally nontoxic, nonimmunogenic, and chemically inert, implants made from these materials may trigger acute and chronic inflammatory responses. These inflammatory reactions may induce degeneration of implanted biopolymer. Interactions between implanted biomaterial and inflammatory cells are mediated by many cellular events involving cellular adhesion and activation. We studied the inflammatory responses in vivo and in vitro to samples of biopolymers composed of poly(ethylene terephthalate-co diethylene glycol terephthalate) plus 0, 5, 25% of polyethylene oxide. We observed that these biopolymers did not induce inflammatory responses when implanted in the peritoneal cavity of mice for 28 days. However we observed deposition of hyaluronic acid at the surface of implanted biomaterial, suggesting that tolerance to biomaterial occurred after surgical implantation. No significant adhesion of inflammatory cells such as mononuclear phagocytes and peripheral leukocytes were observed in vitro, when poly(ethylene terephthalate-co diethylene glycol terephthalate) blends were used as substratum to cellular adhesion. These results suggest that blends composed of poly(ethylene terephthalate-co-diethylene glycol terephthalate) induce low inflammatory cell adhesion, since no rejection of biopolymer was observed when implanted in experimental animal models. PMID- 9870761 TI - Hemostatic capability of rapidly curable glues from gelatin, poly(L-glutamic acid), and carbodiimide. AB - The hemostatic capability of rapidly curable glues composed of gelatin and poly(L glutamic acid) (PLGA) was compared with that of the conventional fibrin glue. The hydrogels produced from mixed gelatin and PLGA aqueous solution within several seconds by addition of water-soluble carbodiimide (WSC) was applied to the dog spleen injured by needle pricking. The WSC-catalyzed gelatin-PLGA glues exhibited higher hemostatic capability than the fibrin glue. The total amount of bleeding from the injured spleen until hemostasis when the gelatin-PLGA hydrogel glues were applied was significantly smaller than that of the fibrin glue application. The gelatin-PLGA glue application enhanced the success rate of complete hemostasis to a significantly greater extent than the fibrin glue, while the frequency of glue applications until achieving complete hemostasis decreased. The gelatin PLGA hydrogels strongly adhered to the surface of dog spleen, whereas the fibrin hydrogel was easily detached from the spleen surface. It was concluded that this strong adhesion mechanically suppressed the bleeding, leading to enhanced hemostasis by the rapidly curable gelatin-PLGA glues. PMID- 9870762 TI - The 2G2 antibody recognizes an acidic 110-kDa human mitochondrial protein. AB - Using fluorescence microscopy, the mouse monoclonal antibody 2G2 was found to label mitochondria in human cells, as assessed by double staining with either Rhodamine 123 or a polyclonal antibody to mitochondrial matrix HSP-60 proteins. No reactivity to the 2G2 antibody was detected in cells from mouse, rat and chicken. Immunoblotting analysis demonstrated that the 2G2 antigen corresponds to a human protein with a relative mobility of 110kDa and an approximate isoelectric point of 6.5 that co-partitions with HSP-60 proteins during isolation of mitochondria from HeLa cells. Close examination of the 2G2 staining pattern in HeLa and Fanconi's anaemia cells revealed differences in the morphology and organization of mitochondria in these two cell types. In HeLa cells, mitochondria appear as individual tubular compartments of variable length and are closely associated with vimentin filaments, particularly at the periphery of the nucleus. In Fanconi's anaemia cells, mitochondria have a filamentous shape and form an interconnected cytoplasmic reticulum running in parallel with both vimentin filaments and microtubules. After stabilization with aldehyde- or alcohol-based fixation protocols that optimize the preservation of cytoskeletal components, the epitope targeted by the 2G2 antibody may serve as a valuable marker in the investigation of relationships between mitochondria and other cellular structures in human cells. PMID- 9870763 TI - Immunolocalization of the vascular endothelial growth factor receptor-2 in the subepicardial mesenchyme of hamster embryos: identification of the coronary vessel precursors. AB - The earliest evidence of the development of the cardiac vessels in mammals is the emergence of subepicardial blood islands, which are thought to originate from mesenchymal progenitors. In order to identify these progenitor cells, we have studied the immunohistochemical localization in the heart of Syrian hamster embryos of the type 2 vascular endothelial growth factor receptor, the earliest molecule known to be expressed in the vasculogenic cell lineage. Only a few immunoreactive subepicardial mesenchymal cells were present by 10 days post coitum. By 11 days post coitum, the subepicardial mesenchymal cells became abundant at the dorsal part of the ventricle, the atrioventricular and the conoventricular grooves. About 20% of cells were labelled with the antibody. Immunoreactive cells were isolated or formed pairs, short cords, rounded clusters or ring-like structures at the subepicardium or, occasionally, within the ventricular myocardium. Other labelled cells were simultaneously cytokeratin immunoreactive. By 12 days post coitum, most immunoreactive mesenchymal cells have been replaced by a capillary network. We propose that an active process of vascular differentiation occurs between 10 and 12 days post coitum in the subepicardium of this species, and it might be a suitable model for the study of vasculogenetic mechanisms. PMID- 9870764 TI - Silver development in microscopy and bioanalysis: a new versatile formulation for modern needs. AB - Potentially, silver development could unify most modern demands for clean, accurately localized marker amplification in microscopy and bioanalysis. However, the existing technology leaves room for improvement in developer design. A new formulation has been devised which, by using principles of silver chelation, avoids problems of self-nucleation and catalysis by light. It is made, just before use, by mixing together equal amounts of stock solutions containing high molarity, Tris-buffered silver nitrate and alcoholic, buffered pyrogallol. The two stocks are easily prepared and have very long shelf-lives. The developer is light insensitive for up to an hour at room temperature, so that development can proceed under ambient light conditions and at the neutral pH most suited to biological systems. The powerful reducer in the suggested formulation should allow the detection of low concentrations of marker signal in a wide range of applications. PMID- 9870766 TI - Transient lectin binding by white matter tract border zone microglia in the foetal rabbit brain. AB - Axonal growth cones of developing white matter tracts are guided through the cerebrum by interactions with cell surface and extracellular matrix molecules expressed by glial cells that mediate cell adhesion and contact-dependent inhibition. Specific carbohydrates are considered essential for the proper functioning of these molecular complexes. We studied developmental aspects of complex carbohydrate expression by white matter glia in the foetal rabbit brain using the tomato lectin Lycopersicon esculentum, which has affinity for components of the extracellular matrix proteins and cell surface proteins (N acetylglucosamine) and activated lysosomal membrane glycoproteins (N acetyllactosamine). Concentrations of the lectin-positive glia were transiently found immediately adjacent to developing white matter tracts of the foetal rabbit brain from 22 to 32 days' gestation. The number of positive cells markedly diminished by the fourth post-natal day and in the adult brain. The lectin positive glia did not react with antibody to glial fibrillary acidic protein. However, they did express the macrophage surface antigen, Mac-1, indicating that the lectin binding reflected the presence of microglial activated lysosomal membranes. These data suggest that, in addition to their role as central nervous system scavengers, microglia are involved in a specifically timed function in the neurodevelopmental programme of white matter tract formation. PMID- 9870765 TI - The expression of transforming growth factor alpha receptor protein and its activation in chicken ovarian granulosa cells of maturing follicles. AB - Epidermal growth factor (EGF) and transforming growth factor alpha (TGF-alpha) are structurally related growth factors that exert their biological actions by binding to the same cell-surface receptor, EGF receptor. However, in chicken cells, human EGF binds with approximately 100-fold lower affinity than human TGF alpha. In a previous study, we localized EGF/TGF-alpha receptor immunohistochemically in the granulosa and theca of the developing follicles of laying hens. We have also shown that TGF-alpha binds to cell-surface receptors of the granulosa cells. The present study characterizes the nature of the EGF/TGF alpha receptor. Immunoprecipitation of receptor proteins from cultured granulosa cells with an anti-EGF receptor antibody (12E) shows the expression of a 170-kDa receptor protein. The expression of the receptor protein decreases with follicular enlargement between the F3 and F1. Incubation of the cells with [125I]TGF-alpha followed by cross-linking with bis(sulphosuccinimidyl)suberate showed that TGF-alpha binds a similar (170 kDa) receptor protein immunoprecipitated with the 12E anti-EGF receptor antibody. The binding of TGF alpha to granulosa cells caused receptor protein oligomerization, yielding the monomeric (170 kDa) and dimeric (340 kDa) protein forms. Oligomerization seemed to favour the formation of the dimeric rather than the monomeric form. Culturing granulosa cells with luteinizing hormone or follicle-stimulating hormone increased the expression of both monomer and dimer forms of the receptor proteins compared with the control. Western blotting analysis with anti-phosphotyrosine antibody revealed that the lysates of TGF-alpha-stimulated cells express phosphotyrosine-containing receptor proteins of 170 kDa and 340 kDa. The results show that chicken granulosa cells express the 170-kDa EGF/TGF-alpha receptor protein, which dimerizes on binding to TGF-alpha, suggesting that the receptor protein may be involved in the signal transduction of TGF-alpha actions in the chicken granulosa cells. PMID- 9870767 TI - Histochemical studies on sialic acids in the epididymis during post-natal development of the rat. AB - A variety of sialic acids contained in the rat epididymis during post-natal development were examined by means of lectin and carbohydrate histochemistry. Epididymides from male Sprague-Dawley rats on post-natal days 14, 21, 30, 39, 49, 56 and 70 were fixed in Bouin's fluid and embedded routinely in paraffin wax. Hydrated sections were subjected either to the lectin methods using biotinylated Sambucus sieboldiana lectin or Maackia amurensis lectin or to the selective periodate oxidation-phenylhydrazine-thiocarbohydrazide-silver protein-physical development technique with or without saponification. The results revealed that sialic acids appeared in the epididymal epithelium at day 14, followed by particular distribution patterns corresponding to cell differentiation during days 21-39. High-level O-acetylation of sialic acids was observed in the principal cells of the initial segment and proximal caput after day 39. These results suggest that sialic acids with different linkages and O-acetylation become adult in distribution at the 'differentiation' period under the influence of androgen, before spermatozoa reach the epididymal lumen. Such carbohydrates may be correlated, at least in part, with sperm-binding sialoproteins, which increase dramatically during the window between days 21 and 39. PMID- 9870768 TI - Evidence for phosphoprotein microspheres in bone. AB - Bone sialoprotein and osteopontin are 'bone-specific' phosphoproteins, but their function is uncertain and their ultrastructural associations remain unclear. Insight into their role was sought by special attention to their general distribution and specific morphology under the high-power optical microscope. Their extracellular staining characteristics were examined in cryosections of adult rat skeletal tissues using two immunohistochemical methods. The two proteins were clearly evident in immature woven bone of endochondral and intramembranous origin (although cartilage was negative, even when calcified). In mature lamellar bone, bone sialoprotein remained ubiquitous, while osteopontin was confined to cement lines and other relatively discrete sites of past and present resorption activity, particularly near blood vessels. In neither case was the distribution of the stain structureless and diffuse. Invariably (except when non-specific), it was sharply defined and had the form of microspheres measuring approximately 1 microm in diameter. In both immature and mature regions, these objects appeared in sheets, chains or groups in a pattern that was evidently coincident with a similar structural arrangement found within the inorganic phase of bone. It was concluded that phosphoproteins are not randomly located throughout the collagenous matrix but are apparently integral to calcified microsphere populations, and it is suggested that these structures are well placed to control the chemical state of the mineral over their surfaces and influence remodelling. PMID- 9870769 TI - Sulphated glycosaminoglycans in guinea pig eosinophils studied by means of cationic colloidal gold. AB - Using bone marrow embedded in hydrophilic resin Lowicryl K4M and cationic colloidal gold pH 1.0 labelling, we studied sites of sulphation and sulphated glycosaminoglycans ultrastructurally in various maturational stages of both eosinophil granulocytes and eosinophil granules of guinea pig. Eosinophil granules reacted positively to cationic gold, the pattern of labelling varying according to the degree of cell maturation. The formation of eosinophil granules takes place throughout the myelocyte stage. Early eosinophil myelocytes contain a large Golgi apparatus with active granulogenesis, while late ones contain a small and less active Golgi apparatus. All the immature granules were labelled positively. However, mature granules with a central crystal bar lost their affinity towards colloidal gold. Interestingly, strong colloidal gold labelling was also observed in the trans to transmost Golgi apparatus, especially in immature eosinophil granulocytes. This indicates that sulphation of glycosaminoglycans occurs in the trans to transmost Golgi apparatus of eosinophil granulocytes. Prior absorption with poly-L-lysine prevented colloidal gold labelling of tissue sections. Methylation of sections at 37 degrees C did not alter the gold labelling, whereas the labelling disappeared after methylation at 60 degrees C. Prior treatment with chondroitinase ABC or heparinase I abolished the majority of colloidal gold labelling in immature eosinophil granules. Taking these results together, we conclude that immature eosinophil granules contain sulphated glycosaminoglycans including chondroitin sulphate or heparan sulphate or both. PMID- 9870770 TI - Thoracoscopy in the evaluation and management of thoracic trauma. AB - Video-thoracoscopy was used to evaluate and manage patients after thoracic trauma. It was used in 29 patients. Indications included retained hemothorax in 16 patients, empyema in 11, evaluation for the source of thoracic bleeding in 1, and an airleak in 1. The mechanism of injury was blunt trauma in 8 cases, 10 with stab wounds, and 11 with gunshot wounds. In blunt trauma, thoracoscopy was carried out an average of 11.7 days post injury, chest tubes were removed after an average of 7 days post thoracoscopy, and discharge averaged 10.7 days after thoracoscopy. The failure rate was 12.5% with no mortality. In stab wounds, it was carried out an average of 8.8 days post injury, chest tube removal occurred after 6.1 days, and discharge averaged 7.8 days after thoracoscopy. The failure rate was 20% with no mortality. In gunshot wounds, it was carried out an average of 7.5 days after injury, chest tubes were removed after 9.9 days, and discharge averaged 16 days post thoracoscopy. The failure rate was 9% with a mortality of 9%. Overall, the failure rate for thoracoscopy was 13.8% (4/29). The mortality rate was 3.5% (1/29). It was successfully performed up to 30 days post injury. It proved to be effective in the management of empyema, evacuation of clotted hemothorax, and diagnosis of ongoing thoracic bleeding. PMID- 9870771 TI - Coronary artery bypass grafting (CABG) after successful percutaneous transluminal coronary angioplasty (PTCA): is PTCA a risk for CABG? AB - We studied patients who underwent a coronary artery bypass grafting (CABG) procedure with previous percutaneous transluminal coronary angioplasty (PTCA). Forty patients had undergone successful PTCA, and required subsequent CABG, between January 1993 and June 1996 (Group I). These patients were matched with 40 patients surgically revascularized without previous PTCA at the same term (Group II). There were no statistical differences among sex, diabetes mellitus, hypertension, family history, smoking, hypercholesterolemia and prior myocardial infarction within the groups. The mean ages were 50.7+/-9.4 and 54.7+/-7.7 years, respectively, in Group I and Group II (P = 0.02). Preoperative mean ejection fraction values were 59+/-5% in Group I and 56+/-7% in Group II (P = 0.01). The mean follow-up period was 21.0+/-9.8 months (1-38 months) for both groups. CABG operations were performed 11.4+/-6.0 months after PTCA. Number of grafts were 2.1+/-0.7 and 2.3+/-0.8 per patient in Group I and Group II, respectively. Mean aortic cross-clamping times were 18+/-3 and 17+/-4 min/graft (P = 0.01) and cardiopulmonary bypass (CPB) times were 34+/-7 and 29+/-7 min for Group I and Group II, respectively, (P = 0.0001). The duration of hospital stay were 9.1+/ 2.5 days for Group I and 8.0+/-1.1 days for Group II (P = 0.008). Freedom from angina at the end of 3 years was 82.5% and 87.5% for Group I and Group II, respectively. One early and two late deaths occured in Group I. One early death and one late death occured in the other group. Survival rates for three years were 92.5% and 95% in Group I and in Group II, respectively. In conclusion, the method of initial revascularization procedure should be considered carefully, as markers of more severe disease may indicate primary CABG and avoidance of an initial PTCA. The initial PTCA may complicate the operation and may increase postoperative morbidity and mortality. PMID- 9870772 TI - Left cerebral hemi-neglect following carotid TEA: diagnostic, pathogenetic and therapeutic considerations. AB - OBJECTIVE: This paper describes the first case report in the literature of a hemi neglect following carotid desobliteration. The disorder is explained in detail and diagnostic as well as pathogenetic aspects are discussed in depth. SUBJECT: Hemi-neglect syndrome is thought to be a rare complication after carotid desobliteration. It describes a neuropsychological disorder which can affect motor, sensory and visual modalities, either singly or in combination. The diagnostic approach is rather simple but requires thorough clinical assessment. It can be caused by embolic or hemodynamic reasons; the clinical findings in either case are highlighted and discussed in detail. CONCLUSIONS: Pre-operative diagnostic evaluation and perioperative management of patients with ICA-stenosis should be undertaken in close co-operation between surgeons and neurologists. This way, the risk of developing neurological complications (i.e. neglect) after surgery can be estimated in the individual case. In high risk cases, indication for surgery should be seen critically. PMID- 9870773 TI - Percutaneous treatment of rare latrogenic arteriovenous fistulas of the lower limbs. AB - Postoperative iliac and popliteal arteriovenous fistulas are extremely rare. This article describes the percutaneous endovascular treatment of iatrogenically induced arteriovenous fistula in two patients with simultaneous use of intravascular ultrasound. In a 61-year-old woman, a fistula between popliteal artery and vein, inadvertently created during implantation of a prosthetic knee joint, was treated with a novel polytetrafluorethylene (PTFE) stent graft. A 48 year-old man underwent endovascular treatment of a large fistula between the right common iliac artery and the left common iliac vein, inadvertently induced during surgery on an intervertebral disc. Congestive heart failure induced by the arteriovenous communication was reduced rapidly. Our results indicate that percutaneous treatment in conjunction with intravascular ultrasound is a useful therapeutic option for vascular lesions. It is less invasive than open vascular reconstruction and has a comparably high success rate. PMID- 9870774 TI - Total versus subtotal thyroidectomy in the management of multinodular goiter. AB - BACKGROUND: The choice between subtotal and total thyroidectomy for multinodular goiter is controversial. MATERIAL AND METHODS: Subtotal and total thyroidectomy have been respectively performed in 108 and 451 euthyroid patients with multinodular goiter. RESULTS: After subtotal and total thyroidectomy, transient recurrent laryngeal nerve (RLN) palsy occurred in 1 patient (0.9%) and 3 (0.6%), transient asymptomatic hypoparathyroidism in 27 (25%) and 131 (29%), and transient symptomatic hypoparathyroidism in 2 (1.8%) and 13 (2.9%), respectively (P = NS). After subtotal thyroidectomy, recurrence occurred in 27 patients (26%) Re-operation in 14 patients resulted in transient RLN palsy in 2 patients (14%), transient asymptomatic hypoparathyroidism in 6 (43 %) and transient symptomatic hypoparathyroidism in 2 (14%). CONCLUSIONS: The failure to demonstrate any 'hormonal advantage' in preserving thyroid tissue by subtotal thyroidectomy, and the low morbidity rate and no need for re-operation after primary total thyroidectomy, make the latter the procedure of choice for the management of non toxic multinodular goiter. PMID- 9870775 TI - Lymph node metastasis and lymphadenectomy for carcinoma in the gastric cardia: clinical experience. AB - METHOD AND MATERIALS: To suggest improved guidelines for treating lymphatic spread and to understand the results of extended lymphadenectomy. A total of 141 cases of cardia carcinoma admitted to our hospital during 1980 to 1993 were analyzed. RESULTS: 123 of 141 cases underwent resection. The incidence of mediastinal lymph node involvement rose from 8.7% in cases with 1-2 cm invasion to 52.9% in cases with more than 4 cm invasion into the esophagus. The incidence of abdominal lymph node involvement also rose with increasing esophageal invasion. Paraaortic lymph node metastasis was frequently observed in cases of more than 1 cm esophageal invasion. D4 lymphadenectomy (extended lymphadenectomy) revealed better results for the cases with lymph node involvement (P<0.05). There was no difference in the incidence of postoperative complication or in operative death, between the abdominal approach and the thoracoabdominal approach, or between the D4 lymphadenectomy and D2 lymphadenectomy. CONCLUSION: Sufficient mediastinal and abdominal lymphadenectomy under a clear wide surgical field achieved through the thoracoabdominal approach appeared to bring better results. PMID- 9870776 TI - Release of immunosuppressive substances after gastric resection is more prolonged than after mastectomy in humans. AB - The effects of gastric and breast cancer surgery on the systemic immune response, and potential correlations with cytokines in peritoneal fluid (PF) and wound fluid (WF) after surgery were investigated in humans. Twenty patients with gastric cancer and 13 patients with breast cancer were studied. Blood natural killer (NK) cell activity and serum soluble interleukin-2 receptor (sIL-2R) were assayed, and prostaglandin E2 and transforming growth factor-beta1 were measured in PF and WF. A prolonged decrease in NK cell activity was observed after gastrectomy compared with mastectomy. Serum sIL-2R levels were increased after gastrectomy, whereas no difference was observed after mastectomy. Cytokine levels in PF persisted on day 1 through 5 after gastrectomy, whereas in WF they dropped sharply on postoperative day 3. The systemic immune response was decreased longer after gastrectomy than mastectomy, and this immune modulation may be partly attributable to the presence of cytokines in local exudates. PMID- 9870777 TI - Laparoscopic cholecystectomy: an analysis on 114,005 cases of United States series. AB - 114,005 cases from 40 United States series of laparoscopic cholecystectomies (LC) were reviewed. Indications, conversion rates, rates of intra-operative cholangiography (IOC), and incidence of bile duct stone and iatrogenic bile duct injuries were assessed. Indications included acute cholecystitis in 11.6% and gallstone pancreatitis in 2.1% of reported cases. Conversion rate was to be primarily related to inflammation. Unsuspected bile duct stones were detected intra-operatively in 7.8% of cases. 561 major bile duct injuries (BDI) and 401 bile leaks (BL) were recorded and acute or chronic inflammation was their most important potential predisposing factor. In series with a high rate of IOC performed during LC, BDJ and BL were slightly lower and lesions recognized intra operatively were much higher than in series with low rate of IOC. BDJ occurred in the first 50 patients of the surgeon's experience in about 91% of the cases. PMID- 9870779 TI - Methotrexate-induced hepatic necrosis requiring liver transplantation in a patient with rheumatoid arthritis. AB - MTX-induced hepatic injury and liver enzyme elevations have been demonstrated after treatment of leukemia, gestational disease and during treatment of psoriasis and rheumatoid arthritis. A 40-year-old man with a long standing history of rheumatoid arthritis was treated with MTX over a 6 month period and developed an overwhelming hepatic necrosis. He was successfully transplanted. PMID- 9870778 TI - The effect of portal hypertension on transforming growth factor-alpha and epidermal growth factor receptor in the gastric mucosa of rats. AB - The aim of this study was to evaluate the expression of transforming growth factor-alpha (TGF-alpha) and epidermal growth factor receptor (EGFR) immunohistochemically in gastric mucosa of portal hypertensive (PHT) and sham operated rats. PHT was induced experimentally by partial ligation of the portal vein (PVL) in 12 male Wistar rats; another group of 12 rats (control) received sham operations. The severity of macroscopic gastric mucosal lesions and immunoreactivity of TGF-alpha and EGFR were evaluated 14 days following the above procedure. The damage to gross gastric mucosa was found to be significantly greater in the PVL group than the control group (p<0.01). Additionally, the TGF alpha and EGFR immunoreactivities were significantly more intense in the PVL groups compared to the control (P<0.05). The TGF-alpha and EGFR immunoreactive cells were more prominent around injured mucosa. From these findings, we suggest that locally produced TGF-alpha and EGFR may play an important role in the gastric mucosal repair following PHT to rats. PMID- 9870781 TI - Primary lymphoma of the liver: a case report and review of the literature. AB - Primary lymphoma of the liver (PLL) is a rare disease and estimation of the real number of cases may be difficult because strict diagnostic criteria for the definition of a primary lesion are quite often not followed. We report here on a case of a patient affected by PLL who underwent successful surgical resection of the lesion followed by chemotherapy and autologous bone marrow transplantation. The patient is alive and disease free 62 months after resection. PMID- 9870780 TI - Protecting the viability of hepatic allografts procured from non-heart-beating donors by blockade of endothelin and platelet activating factor in porcine liver transplantation. AB - PROBLEM: The function of hepatic allograft from non-heartbeating donors (NHBD) is significantly affected by warm ischemic injury before harvesting. MATERIALS AND METHODS: The effects of the endothelin (ET) antagonist TAK-044 and the platelet activating factor (PAF) antagonist E5880 on the function of grafts from NHBD were evaluated in a porcine orthotopic liver transplantation. The liver grafts were subjected to 90 min of warm ischemia and 4 hours cold preservation. Group 1 (n = 4; n is the number of donor/recipient pairs) was used as a control (untreated). In group 2 (n = 4), donors and recipients were treated with TAK-044 (3 mg/kg). In group 3 (n = 4), pigs were treated with E5880 (0.3 mg/kg). In group 4 (n = 4), pigs were treated with TAK-044 and E5880. RESULTS: The 7 day survival rate of the recipients in groups 1, 2, 3 and 4 was 0, 25, 25 and 100%, respectively (p<0.05, group 1 versus 4). The increases of the serum concentrations of aspartate transaminase, lactate dehydrogenase and arterial lactate 1-4 hours after transplantation were significantly inhibited in the treated groups. CONCLUSION: The blockade of ET and PAF has protective effects on the function of hepatic grafts from NHBD. PMID- 9870782 TI - Long-term results of corticosteroid administration via appendicostomy in patients with ulcerative colitis involving the entire colon. AB - BACKGROUND: Ulcerative colitis (UC) of an unknown etiology frequently demonstrates repeated active and inactive stages. As a result, it is difficult to sustain long-term remission under conservative therapy. METHODS: Ten patients who presented suffering from UC involving the entire colon were treated. All patients had been previously treated at other hospitals usually with sulphasalazine and either corticosteroids or steroid enemas for the primary complaints of muco bloody stool or frequent diarrhea. All patients underwent either an appendicostomy or cecostomy, and were injected with dexamethasone via an artificial fistula twice a day. RESULTS: This treatment led to remission, and, as a result, the symptoms of anal bleeding or muco-bloody stools disappeared in all patients. Radiological, endoscopic, and pathological studies revealed a dramatic response to steroid injection from the fistula. The mean follow-up period was 44.7 months (3-122 months). Eight patients remained free from any symptoms of UC. One underwent a proctocolectomy because of side effects due to steroid treatment. The other patient died suddenly of unknown causes. The mean symptom-free period after cecal injection was 39.5 months (1-119 months). The mean proportion of disease free period from UC, compared with the total follow-up period after surgery, was 88%. After remission, eight patients were able to return to a normal lifestyle at home using a peritoneal button. They could also take a bath and continue their school or social lives in almost the same way as healthy persons. CONCLUSIONS: We thus recommend this new, minimally-invasive therapy for patients with UC involving the entire colon who demonstrate resistance to conventional conservative therapy. As a result of such treatment, all patients were able to achieve a comfortable lifestyle after undergoing minimally-invasive surgery. PMID- 9870783 TI - Outcome after emergency subtotal/total colectomy compared to elective resection in patients with left-sided colorectal carcinoma. AB - Long term survival of 72 patients who underwent an emergency one-stage subtotal/total colectomy with primary anastomosis for obstructing carcinoma of the left colon was compared with 141 patients who underwent an elective procedure for non-obstructing carcinoma of the same location. Patients who presented with intestinal obstruction were older than electively treated patients (68.5 versus 62.1 years), postoperative mortality was higher (13.8% versus 7.8%). 5-year survival rate of patients treated by R0 emergency subtotal/total colectomy was comparable to electively R0 resected patients (69% versus 61%). There was no significant difference in survival comparing the same Dukes' stage between the two groups. One-stage procedure could be regarded as a preferred method of treatment in patients with obstructing carcinoma of the left colon, either as a subtotal/total colectomy in cases when the colonic wall is endangered or as a resection with primary colo-colonic anastomosis after on-table lavage. PMID- 9870785 TI - Hydroxyethyl starch solution: is it a new alternative way of treatment in bacterial translocation? AB - In this experimental study, the effects of mannitol and hydroxyethyl starch solution (HES) on bacterial translocation were evaluated in a rat model of simple mechanical bowel obstruction. The mechanical bowel obstruction was created by ligation of the ileum 1 cm proximal to the ileocecal valve. Excluding the control group (n = 7), the rats were given 2 ml of mannitol or 2 ml of 6% hydroxyethyl starch solution intraperitoneally during the procedure. Relaparotomy was made and segmental ileal resection, total mesenteric lymph node excision and removal of tissue sample of liver were performed 24 h after the mechanical bowel obstruction, both for histopathological and microbiological examination. The rates of colony formation in the mesenteric lymph nodes after the mechanical bowel obstruction were 71% (5/7) in control animals, 43% (3/7) in animals given mannitol and only 14% (1/7) in animals given HES. In the liver samples observed for the bacterial translocation, the corresponding figures were 14% (1/7), 28% (2/7) and 0% (0/7) respectively. The difference between hydroxyethyl starch solution and control groups were statistically significant (P<0.04). The bacterial population/g tissue obtained from intraluminal bacterial cultures of the ileum was also depressed in hydroxyethyl starch solution group when compared to the control (P<0.03). As a result, we confirmed that hydroxyethyl starch solution, a plasma volume expander, depressed the bacterial translocation to the mesenteric lymph nodes and liver and also reduced intraluminal bacterial overgrowth in the simple mechanical bowel obstruction model of rats. We believe that hydroxyethyl starch solution may be an alternative therapeutic agent for the pre-operative fluid resuscitation in simple mechanical bowel obstruction. PMID- 9870784 TI - Immobilized proteinases in the treatment of diffuse purulent peritonitis. AB - In vitro experiments on fibrin films using purulent exudate from the abdominal cavity of rats with experimental peritonitis demonstrate the fibrinolytic effect of bacterial proteinases immobilized on a polymeric matrix. The application of Imozimaza in the complex treatment of experimental peritonitis by the way of intraperitoneal lavage resulted in reliable lowering of mortality, due to the lysis of fibrinopurulent abdominal contents and better contact between antibacterial agents and peritonitis pathogens. In the clinic, prolonged abdominal proteolysis was applied to 44 patients with postoperative diffuse purulent peritonitis of >24 h duration. Under the conditions of programmable relaparotomy, intraperitoneal Imozimaza infusion led (as in in vitro tests) to the lysis of fibrinopurulent masses, which contained micro-organisms of an order higher than exudate. It was accompanied by increase in the drainage efficacy, absence of fragmentation of abdominal contents and absence of secondary abscesses. The use of Imozimaza on the background of complex antibacterial treatment and combined homeostatic therapy resulted in lowering of mortality from 65.8% to 27.3%. Complications and contra-indications for Imozimaza use in diffuse purulent peritonitis were not registered. PMID- 9870786 TI - Laparoscopic inguinal hernia repair. AB - Open hernia repair is associated with significant postoperative pain and disability resulting in delayed return to full activity. Laparoscopic hernia repair has been advocated as the procedure that combines the benefit of tension free repair with the preservation of the basic anatomy of the inguinal area. We present our experience with 803 laparoscopic hernia repairs in 517 patients over a period of 66 months (August 92 to February 98). The effects of the learning curve and the refinement of the technique had their impact on earlier results and complications. However, with more experience we found that the laparoscopic preperitoneal approach is safe and efficacious. There was no mortality. Most patients (85%) were discharged home within 24 h of the procedure and returned to full activity within 10 days. Patient satisfaction was excellent. The complication rate decreased and operative time was reduced with experience. This procedure is clearly indicated in patients who have recurrent or bilateral hernias. It is associated with shorter convalescence and a quick return to work. PMID- 9870787 TI - Treatment of urinary incontinence with AMS 800 artificial urinary sphincter. AB - BACKGROUND: Urinary incontinence has a significant impact on the quality of life for patients. The introduction of the AMS 800 artificial urinary sphincter by American Medical Systems (Minnetonka, MN, USA) has revolutionized the treatment of urinary incontinence due to pure sphincteric incompetence. PATIENTS AND METHODS: From April 1989 to October 1997, a total of 52 patients (42 men and 3 women) underwent implantation of the AMS 800 artificial sphincter. Patient age ranged from 31 to 87 years (mean 69.5 years). They all were suffering from urinary incontinence caused by sphincteric damage, always of iatrogenic origin. Follow-up was 5-110 months, with a mean of 35 months. RESULTS: The restoration of continence was complete for 35 patients and allowed them all to return to their normal life. 7 of the patients who had a successful implant died because of other problems. For 2 patients follow-up was not available. Explantation was performed 8 times: in 5 male patients the device was removed because of periprosthetic infections; in 1 female patient with ileal neobladder, because of serious urethral necrosis; and in 2 male patients because of an urethral erosion. CONCLUSIONS: The AMS 800 artificial urinary sphincter is the most efficacious treatment currently available for urinary incontinence due to pure sphincteric incompetence. Implanted patients achieved acceptable urinary control. The implantation is associated with 2 distinct complications: surgical and mechanical. PMID- 9870788 TI - Extraperitoneoscopic colposuspension using CO2 distension method. AB - PURPOSE: To validate our experience in extraperitoneoscopic colposuspension for genuine stress incontinence. MATERIALS AND METHODS: Between March 1995 and July 1996, 48 women who had genuine stress incontinence underwent extraperitoneoscopic colposuspension in our institute. After standard laparascopic surgery preparation, a 10 mm puncture site was made midline just in the cm above the pubic hair line, and the extraperitoneal space was developed with the higher pressure of insufflating CO2. A pair of sutures was inserted at the level of the midurethral and unrethrovesical junction with Cooper's ligament. RESULTS: All of these patients underwent the same procedures. The average blood loss was less than 50 ml, with a range from 10 to 200 ml. The operative time was from 20 to 90 min, with a mean time of 32 min. There was one bladder injury, 2 cases of voiding difficulties and 2 of detrussor instability in our series and the overall complication rate was 10.4%. So far, 45 of the 48 patients are satisfied with the surgery. CONCLUSIONS: Laparoscopic Burch colposuspension is a practicable surgical procedure for managing stress incontinence. Extraperitoneal space was created easily with the higher pressure of insufflating CO2. An extraperitoneoscopic approach can reduce the necessity of laparscopic suturing. Moreover, it avoids violating the peritonel cavity and reduces the potential risk of postoperative adhesion formation and the discomfort resulting from pneumoperitonium. Thus, extraperitoneal colposuspension affords an alternative to laparoscopic or abdominal retropubic colposuspension in well-selected patients. PMID- 9870789 TI - Prognostic factors in patients with with locally advanced cervical cancer treated with radical hysterectomy and adjuvant radiotherapy. AB - The purpose of this study was to investigate the presence of risk factors for node metastases and to estimate survival in patients with cervical cancer, stages IB and IIA. In a retrospective study of 103 patients with cervical cancer stages IB and IIA, all treated with radical hysterectomy and adjuvant radiotherapy, we estimated survival curves according to different prognostic parameters. Mean follow-up time was 97 months. A significant difference between clinical pre operative assessment and histological determination of real extent of the disease was evidenced. Pelvic lymph node metastases (P = 0.0005) significantly correlated with survival. This study shows that only lymph node involvement is an independent prognostic factor. Stage acts through nodal status in its impact on survival. A surgical-pathological staging in early stage cervical cancers is found to be more appropriate to correctly estimate patients' survival and prognosis. PMID- 9870790 TI - Evaluation of the stability of pin configuration in K-wire fixation of displaced supracondylar fractures in children. AB - Precutaneous Kirschner wire fixation of displaced Gartlands types II and III supracondylar fractures with image guidance remains a preferred procedure for most orthopaedic units. Various K-wire configurations have been in use, but recent objective evaluation of pin stability has favoured two techniques; the 2 crossed pins placed from the medial and lateral epicondyles, and 2 parallel lateral pins when the former technique is impracticable because of swelling. We reviewed patients who had K-wire fixation precutaneously over a 2-year period (Feb 1996 to Feb 1998). There were 44 children, 30 males, 14 females, ages between 1-15 years. 19 patients had (medial and lateral pins) 15, (2 parallel lateral) and 10 patients (2 lateral crossed) precutaneous pin placement respectively. Their postoperative course indicated that maximum stability was obtained with two opposite crossed pins, followed by the 2 paralleled pin method. These observation determined the choice of technique by the surgeons who operated on these patients. PMID- 9870791 TI - Mast cells and gastric mucosal damage: yet another mechanism of injury? PMID- 9870792 TI - Mucobilia: the clinical spectrum, diagnosis and treatment. PMID- 9870793 TI - Treatment of Helicobacter pylori infection to reduce gastric cancer incidence: uncertain benefits of a community based programme in Australia. AB - Helicobacter pylori is a cause of gastric adenocarcinoma, but the role of H. pylori eradication in reducing cancer risk is unknown. We sought to estimate the benefits of a screening and treatment programme for H. pylori infection, aimed at reducing the incidence of gastric cancer in Australia. The impact of this programme on cancer incidence was evaluated in sensitivity analyses utilizing Western Australian Cancer Registry data and published data on the epidemiology of H. pylori and gastric cancer. The impact of variation in parameters used in the sensitivity analyses was substantial, ranging from a 38% reduction in lifetime risk of gastric cancer in a best-case to 3% in a worst-case scenario. In an intermediate-case scenario there is a 23% reduction in lifetime risk, but in real terms this reflects a fall in cumulative incidence from 0.7 to 0.5% for males or 0.3 to 0.2% for females. The projected cumulative lifetime incidence of gastric cancer in H. pylori-infected males is 2.2% and 0.9% for females; this contrasts with 0.4 and 0.2%, respectively, for those never infected. According to an intermediate-case scenario, to prevent one gastric cancer, screening with or without subsequent treatment would be required in 617 men or 1639 women. Furthermore, this programme may be less effective in reducing cancer incidence than would be achieved naturally over the next 15 years, providing the current annual decline in gastric cancer incidence continues. In conclusion, the benefits of a community based programme of H. pylori eradication in terms of cancer risk reduction remain unclear, related largely to uncertainties in the parameters used to calculate these benefits. In Australia, any benefits obtained are likely to be, at best, modest. PMID- 9870794 TI - Survival of Helicobacter pylori in milk and tap water. AB - The survival capacity of Helicobacter pylori in artificially contaminated milk and tap water was investigated in the study. Helicobacter pylori could survive for up to 10 days in milk at 4 degrees C storage but only 4 days in tap water with a steady decrease of colony forming units. However, electron microscopy clearly showed that the non-culturable coccoid form was present in tap water which had been kept at 4 degrees C for 7 days. It is concluded that H. pylori may survive in tap water as well as in milk, with the implication that they may, thereby, act as a vehicle of transmission. PMID- 9870795 TI - Gastric mucosal damage induced by compound 48/80: roles of serotonin and nitric oxide. AB - The roles of nitric oxide (NO) and serotonin (5-HT) in the development of gastric mucosal lesions induced by compound 48/80 (48/80) were investigated in rats. Repeated i.p. administration of 48/80 (1 mg/kg) produced damage in the stomach with severe oedema in the submucosa. The lesions induced by 48/80 were prevented by FPL-52694 (a mast cell stabilizer) and methysergide but not tripelennamine. The lesions were also inhibited by simultaneous administration of N(G)-monomethyl L-arginine (L-NMMA), and this effect was mimicked by inducible NO synthase (iNOS) inhibitors, such as aminoguanidine or dexamethasone and significantly antagonized by coadministration of L-arginine. The mucosal myeloperoxidase activity, thiobarbituric acid reactants and vascular permeability in the stomach were all increased after 48/80 treatment and the changes were also attenuated by cotreatment with L-NMMA. Repeated s.c. treatment with 5-HT (20 mg/kg) provoked similar gastric lesions, which were also prevented by methysergide and iNOS inhibitors, as well as antioxidative drugs, such as allopurinol (a xanthine oxidase inhibitor) and hydroxyurea (a neutrophil-reducing agent). The Ca2 independent NO synthase (NOS) activity was increased in the gastric mucosa after administration of 48/80 or 5-HT and this change was inhibited by dexamethasone. These results suggest that: (i) the repeated administration of 48/80 induced inflammatory gastric lesions in the rat stomach, mediated by endogenous 5-HT; (ii) NO/iNOS is involved in the pathogenic mechanism of 48/80-induced gastric lesions, in addition to oxyradical formation; and (iii) the deleterious role of NO in this lesion model can be accounted for by a cytotoxic action of peroxynitrite that is formed in the presence of superoxide radicals. PMID- 9870796 TI - The recto-anal inhibitory reflex: abnormal response in diabetics suggests an intrinsic neuroenteropathy. AB - As electrical stimulation of the rectum has been shown to result in reflex internal sphincter inhibition mediated by intrinsic nerves, we aimed to evaluate the integrity of these nerves in the rectum of diabetic patients. Anal canal pressure, recto-anal inhibitory reflex (RAIR) and continence were evaluated in 30 diabetic patients (male:female 13:17, median age 57 years, range 37-70) and these data were compared with similar data obtained from 22 age- and sex-matched healthy controls (male:female 9:13, median age 51 years, range 19-65 years). Median duration of diabetes was 8 years (range 3-30). Twelve (40%) of the 30 diabetics had impaired continence for gas (n = 12) and liquid faeces (n = 3). None of the controls had incontinence. Median maximum resting anal canal pressure (MRP) was: patients 30 mmHg (range 20-75 mmHg) versus controls 40 mmHg (range 20 105 mmHg, P = 0.61). Median maximum squeeze pressure (MSP) was 65 mmHg (range 30 150 mmHg) in patients versus 84mmHg (range 35-230 mm Hg) in controls (P = 0.59). Median threshold rectal mucosal electrosensation (RMES-T) was 27mA (5-40 mA) in patients versus 13 mA (5-28 mA) in controls (P = 0.03). Maximum tolerable rectal mucosal electrosensation was 40 mA (20-60) in patients versus 20 mA (10-30), in controls (P = 0.042, all comparisons using the Wilcoxon rank test). Recto-anal inhibitory reflex was present in eight, abnormal in five (one incontinent) and absent in 17 (11 incontinent) diabetics, while it was present in 18 and abnormal in four controls (test of proportion, P = 0.031). Blood glucose in diabetics on the day of the procedure was 98 mg/dL (70-165 mg/dL). Rectal mucosal electrosensitivity and RAIR were impaired in significantly more patients with diabetes than controls, implying impairment of intrinsic neuronal function. The recto-anal inhibitory reflex was either impaired or absent in all diabetic patients with incontinence. PMID- 9870797 TI - Multiple truncated transcripts of TSG101 in gastrointestinal cancers. AB - The candidate tumour suppressor gene TSG101, located on chromosome 11p15, has been associated with frequent intragenic deletion in uncultured primary human breast cancers. Using paired tumour and normal tissues from surgical specimens, we performed nested reverse transcriptase-polymerase chain reaction and direct sequencing to analyse TSG101 exons 1-6 from 32 gastric, 30 colorectal and 16 oesophageal cancers. Truncated transcripts, were found in both tumour and normal tissues from the stomach (15.6 and 12.5%), colon (13.3 and 3.3%) and oesophagus (25 and 25%). Multiple truncated transcripts in individual specimens were also observed. Two types of splicing patterns, one with three to six bases homology at the deletion junction (25.9%), the other with donor site 5' GT and acceptor site 3' AG (55.6%), were the most common patterns. We conclude that in gastrointestinal cancers, truncated transcripts of TSG101 gene occur not uncommonly and do so with a specific splicing pattern. PMID- 9870798 TI - Antibiotic-associated haemorrhagic colitis. AB - Antibiotic-associated haemorrhagic colitis is an uncommon cause of bloody diarrhoea in patients taking penicillin or penicillin-related antibiotics. Symptoms of abdominal pain and bloody diarrhoea occur within 1 week of antibiotic use and resolve without specific therapy within days of discontinuing the offending antibiotic. There is an apparent increased incidence of the disease in patients of Oriental ethnicity. The pathogenesis is unknown. We present two cases of haemorrhagic colitis in patients taking penicillin-related antibiotics who presented within 4 months of each other. One of the patients was being treated for Helicobacter pylori infection. The published literature is reviewed with particular emphasis on the histology and pathogenesis of the condition. PMID- 9870799 TI - Inflammatory bowel disease in Saudi Arabia: presentation and initial management. AB - Non-specific Inflammatory Bowel disease (IBD) is infrequently seen in the Arabs of the Arabian Peninsula. This paper documents the presentation and initial management of 101 such Arabs treated for inflammatory bowel disease between 1976 and 1994. Medical records were examined and patients were classified according to the Organisation Mondiale de Gastroenterologie diagnostic scoring system. Sixty seven patients had mucosal ulcerative colitis, 28 Crohn's disease (CD) and six indeterminate colitis; age range 2-71 years. Three patients had a family history of (IBD). The diagnosis was made by a combination of the patients' history, physical, radiological, endoscopic and histological examination; however, eight patients were not examined endoscopically and one in four patients was not biopsied at presentation. Thirty-nine patients did not receive any treatment prior to referral. One in three patients was first treated as inpatients. Giardia, Amoeba, Salmonella, Shigella or Schistosoma were detected in the stools of 36 patients at presentation. Following appropriate treatment, these infections were eliminated but the patients' symptoms persisted. Six patients who were thought to have gastrointestinal tuberculosis were subsequently diagnosed with CD. Ten patients had extraintestinal manifestations of their IBD and 11 had side effects (osteoporosis, Cushingoid features and growth retardation) from steroid treatment at other hospitals. Diagnosis was often delayed because infectious diarrhoea was common and awareness was low. Patients were referred late and some had developed complications of therapy. PMID- 9870800 TI - Association of mutations in the core promoter and precore region of hepatitis virus with fulminant and severe acute hepatitis in Japan. AB - It was recently reported that mutations in the precore and core promoter region of hepatitis B virus (HBV) are associated with fulminant hepatitis. The aim of this study was to investigate the association of mutations in the precore and core promoter region of HBV with fulminant and severe acute hepatitis. We studied Japanese patients with acute HBV infection, including seven patients with fulminant hepatitis, 12 with severe acute hepatitis and 41 with acute self limited hepatitis. The presence of HBV mutants was examined by using a point mutation assay to detect a G to A transition at position 1896 in the precore region and an A to T transition at position 1762 and a G to A transition at position 1764 in the core promoter region. Significant differences in the proportion of mutations in the precore or core promoter region were present between patients with fulminant hepatitis and self-limited acute hepatitis (7/7 (100%) vs 4/41 (9.8%), P<0.01) and between severe acute hepatitis and self limited acute hepatitis (6/12 (50.0%) vs 4/41 (9.8%), P<0.01). The frequency of mutation increased proportionately with the severity of disease in patients with acute HBV infection. Fulminant hepatitis B in Japan is closely associated with mutations in the core promoter and precore gene of HBV. Point mutation assays for HBV precore and core promoter analysis may be useful to predict the outcome of liver disease in patients with acute HBV infection. PMID- 9870801 TI - Fibrosing cholestatic hepatitis after living related-donor renal transplantation. AB - A 43-year-old man underwent living related-donor renal transplantation because of chronic renal failure in 1991. During the transplant period, both donor and recipient were seronegative for hepatitis B surface antigen (HBsAg). The donor was seropositive for antibody to hepatitis B surface antigen (anti-HBs) due to hepatitis B virus (HBV) vaccination. After transplantation, FK506 and methylprednisolone had been administered to the patient as immunosuppressants. In 1993, HBsAg appeared in his serum. His alanine aminotransferase level elevated gradually during 1995 and then in 1996, general fatigue, ascites and jaundice developed. At this time his serum was positive for hepatitis B e antibody, contained more than 100000 Meq/mL HBV-DNA and 100% precore mutant. Despite subsequent intensive therapy, liver dysfunction progressed and this patient died of hepatic failure 2 months following admission. At autopsy, the liver exhibited cholestasis, fibrosis extending from the portal tracts, mild inflammation and hepatocytes with a ground-glass appearance. In addition, HBsAg and hepatitis B core antigens had accumulated in the hepatocytes. Consequently, the final diagnosis was fibrosing cholestatic hepatitis (FCH) due to precore mutant HBV infection contracted after renal transplantation. It is unclear when and where the recipient liver became HBV infected. Nevertheless, after renal transplantation, while receiving immunosuppressive drugs, HBV appeared to have the potential to cause hepatic failure and FCH may have been a fatal complication for the recipient. PMID- 9870802 TI - Evidence of transmission of hepatitis B virus to spouses from sequence analysis of the viral genome. AB - Heterosexual contact is one of the common routes of transmission for hepatitis B virus (HBV) among adults in Taiwan, but only a few studies have provided direct evidence at the level of the HBV genome of infected couples with acute non fulminant hepatitis to document a common source. By cloning and sequencing polymerase chain reaction-amplified HBV-DNA, we analysed the sequences of the conserved region of the surface gene (nucleotide (n.t.) 305-513, representing 6.5% of the viral genome) of HBV in five HBV-infected index patients, their spouses and four randomly selected HBV carriers as controls. Risk factors associated with acute HBV infection in index cases were sexual contact with their spouses within 3 months before the onset of hepatitis. For all five couples, the HBV-infected index patient and the spouse shared a 100% sequence homology of HBV DNA. In contrast, there was significantly more variation (mean heterogeneity 6.1%, range 1-13.9%) in the amplified region between the five couples and between each couple and the controls (P<0.001). This study demonstrated that sequence analyses can correlate well with epidemiological findings and confirm the value of the molecular approach for linked infections of HBV through heterosexual contact between spouses. Susceptible adults should receive vaccination. PMID- 9870803 TI - Radiotherapy in the treatment of duodenal bleeding due to hepatocellular carcinoma invasion. AB - Haemorrhage from an hepatocellular carcinoma (HCC) directly invading the gastrointestinal (GI) tract is uncommon. A 58-year-old man was admitted with upper gastrointestinal (UGI) bleeding and panendoscopy on examination revealed a large duodenal ulcerative bleeding mass. The mass was eventually diagnosed as HCC by pathological examination. The bleeding failed to respond to conventional management of haemostasis, but resolved with an external beam of radiotherapy with a total dose of 6000 cGy over a 5 week period. This unusual presentation of UGI bleeding, due to HCC invading the duodenum and treated by radiotherapy, has not been previously reported. PMID- 9870804 TI - Imaging of follicular dendritic cell tumours of the liver. AB - Follicular dendritic cell tumour of the liver is a recently recognized entity. To date, only two cases have been described, both in the pathology literature. Histologically, it resembles an inflammatory pseudotumour and immunohistochemical and ultrastructural studies are required for its diagnosis. The ultrasound, computed tomography and angiographic features of two cases of follicular dendritic cell tumour of the liver are described in detail. One of the patients had multiple recurrences of this tumour. The imaging features are very similar to those of hepatocellular carcinoma. As follicular dendritic cell tumour is considered to be of low-grade malignant potential, in contrast to the dismal prognosis for hepatocellular carcinoma, it is important to be able to accurately distinguish between the two types of tumour prior to initiating definitive therapy. PMID- 9870805 TI - Power Doppler imaging of hepatic tumours: differential diagnosis between hepatocellular carcinoma and metastatic adenocarcinoma. AB - We evaluated the usefulness of power Doppler imaging in the differential diagnosis between hepatocellular carcinoma and metastatic adenocarcinoma. Forty seven patients with hepatocellular carcinoma and 18 patients with metastatic adenocarcinoma were evaluated using power Doppler imaging. The colour signals of hepatic tumours were graded as follows: 1, colour signals only in the marginal area; 2, small dot or dotted line colour signals within the tumours; 3, continuous solid line colour signals within the tumours. The grade 3 colour signals were classified in the following three patterns; winding line pattern, stretched line pattern and mixed pattern. The colour signals of hepatocellular carcinoma were grade 1 in seven patients, grade 2 in 11 and grade 3 in 29. The colour signals of metastatic adenocarcinoma were grade 1 in three patients and grade 3 in 15. Of the 29 hepatocellular carcinoma patients with a grade 3 signal, 26 patients had winding line patterns and three had mixed patterns. Of the 15 metastatic adenocarcinoma patients with a grade 3 signal, 12 patients had stretched line patterns and three had mixed patterns. In conclusion, power Doppler imaging is useful in the differential diagnosis between hepatocellular carcinoma and metastatic adenocarcinoma to evaluate the colour signal pattern within the tumour. PMID- 9870807 TI - Hepatobiliary and pancreatic imaging: abdominal pain and a pancreatic lesion. PMID- 9870806 TI - The comparative potency of cholesterol crystallization-effector proteins in supersaturated model bile systems: association with vesicle transformation. AB - Various proteins which affect cholesterol crystallization are known to be present in bile, although the relative potency of their action is yet to be established. In this study, we evaluated the comparative potency of nucleating-effector proteins using a recently developed method for quantitative assessment of vesicle transformation in supersaturated model bile systems, to partially characterize mechanisms of their action. Concanavalin A-bound glycoproteins isolated from human gall-bladder bile shortened cholesterol crystallization time by 40% and increased cholesterol growth rate and final crystal mass by 161 and 19%, respectively, when compared to the control. In addition, immunoglobulins isolated from human gall-bladder bile increased cholesterol growth rate by 9%, but showed no significant effect on cholesterol crystallization time and final crystal mass. In contrast, human serum apolipoproteins A-I and B reduced cholesterol growth rate by 26 and 31% and reduced final crystal mass by 12 and 21%, but did not affect cholesterol crystallization time. Gel permeation chromatography revealed that proteins were distributed to both vesicles and bile salt micelles, but that no marked redistribution of lipids was caused by addition of these proteins. Furthermore, no significant difference in crystal structure was observed by video enhanced contrast microscopy. These results indicate that nucleating-effector substances tested in this study may modulate vesicular cholesterol-holding capacity, thus affecting cholesterol crystallization. Such modulation is based upon the protein-vesicle association which defines the physico-chemical metastability of vesicular cholesterol. PMID- 9870808 TI - Gastrointestinal: Cronkhite-Canada syndrome. PMID- 9870809 TI - Who is a physician? 1964. PMID- 9870810 TI - A combined microsurgical skull-base and endovascular approach to giant and large paraclinoid aneurysms. AB - BACKGROUND: The treatment of giant and large paraclinoid aneurysms remains challenging. To improve exposure, facilitate the dissection of aneurysms, assure vascular control, reduce brain retraction and temporary occlusion time, enable simultaneous treatment of associated lesions, and achieve more successful treatment of "difficult" (atherosclerotic and calcified) aneurysms, we combined the skull-base approach with endovascular balloon occlusion of the internal carotid artery (ICA) and suction decompression of the aneurysm. METHODS: Sixteen female patients were treated, eight with giant aneurysms and eight with large aneurysms. Eight aneurysms occurred on the right side and eight on the left. Eight patients had an additional aneurysm; five were clipped during the same procedure. Three patients had infundibular arterial dilation. One patient had an associated hemangioma of the ipsilateral cavernous sinus. The cranio-orbital zygomatic approach was used for all patients. The anterior clinoid was drilled, and the optic nerve was decompressed, dissected, and mobilized. Transfemoral temporary balloon occlusion of the ICA in the neck was followed by placement of a temporary clip proximal to the posterior communicating artery. Suction decompression was then applied. All aneurysms were then successfully clipped, except one that had a calcified neck and wall that could not be collapsed. Intraoperative angiography performed in 13 of 15 patients with clipped aneurysms confirmed obliteration of the aneurysm and patency of the blood vessels. RESULTS: Postoperative results were good in 14 patients. One patient had right-sided hemiplegia and expressive aphasia, which improved after rehabilitation. One patient with an additional basilar tip aneurysm clipped simultaneously died on the fifth postoperative day because of intraventricular hemorrhage. The origin of bleeding could not be determined on autopsy. Surgical difficulties and morbidity stemmed mainly from a severely calcified or atherosclerotic aneurysmal neck. CONCLUSION: The combination of skull-base approaches and endovascular balloon occlusion coupled with suction decompression is a successful option for the treatment of these challenging aneurysms. PMID- 9870811 TI - Complications of subcutaneous low-dose heparin therapy in neurosurgical patients. AB - BACKGROUND: Venous thromboembolism is a major cause of postoperative morbidity and mortality in neurosurgery. The use of low-dose unfractionated heparin therapy perioperatively for prophylaxis against deep vein thromboses and pulmonary embolism has been well demonstrated in many other surgical specialties but is less commonly used in neurosurgery because of fears of devastating postoperative hematomas. METHODS: The safety of such therapy has been analyzed in 950 patients undergoing an inpatient neurosurgical procedure. 872 patients (152 cranial procedures) completed treatment with 5000 U sodium heparin subcutaneously twice a day, commencing before surgery and continuing till patients were ambulatory. RESULTS: There were three minor hemorrhagic complications-two superficial wound hematomas (one requiring treatment) and one gastrointestinal hemorrhage identified. Three clinically significant major complications developed, two epidural hematomas after spinal surgery requiring evacuation and one intraventricular hemorrhage after brain biopsy. CONCLUSION: This report, along with an analysis of previously published reports of low-dose perioperative heparin therapy in neurosurgical patients, suggests that such therapy is unlikely to be associated with increased morbidity. Given the known efficacy of low-dose heparin in reducing venous thromboembolism in other surgical patients, such therapy may reduce mortality and morbidity from thromboembolic complications in neurosurgical patients with minimal risk. PMID- 9870813 TI - Location of the pain: lumbar spine. PMID- 9870812 TI - Recombinant tissue plasminogen activator for the treatment of spontaneous adult intraventricular hemorrhage. AB - BACKGROUND: Intraventricular hemorrhage (IVH) has a poor prognosis with mortality rates of between 80 and 100% when all four ventricles are involved. Fibrinolytic therapy has been reported to improve overall outcome. METHODS: Patients with severe primary IVH were treated by direct intraventricular injection of recombinant tissue plasminogen activator (rt-PA) into the lateral ventricles, followed by cerebrospinal fluid (CSF) drainage if the intracranial pressure rose above 20 mm Hg. RESULTS: Over a 15-month period from 1995 through 1996, 10 patients were treated, (4 male and 6 female, mean age 35 years; range, 21-55 years). The mean Glasgow Coma Scale score on admission was 6 (range, 4-8) and the mean Graeb score for severity of IVH on the first CT scan was 10 (range, 8-12). Angiography was negative in five cases but identified arteriovenous malformations in three, a post-traumatic pseudoaneurysm in one, and Moya-moya disease in one. The mean total dose requirement of rt-PA was 8.25 mg (range, 6-12 mg) with a significant reduction in the mean Graeb score after 7 days to 3.9 (range, 2-7, p<0.0001). Outcome at 3 months was death in one case (mortality 10%), severe disability in two (20%), moderate disability in three (30%), and good result in four (40%). Four patients (40%) required subsequent CSF shunting. No complications of rehemorrhage, infection, or catheter obstruction were encountered. CONCLUSION: Intraventricular fibrinolysis with rt-PA seems to be safe and effective for the treatment of severe IVH. PMID- 9870814 TI - Vertebral hemangiomas: diagnosis, management, natural history and clinicopathological correlates in 86 patients. AB - BACKGROUND: Vertebral hemangiomas are characterized by diverse clinical histories, radiological features, and results of surgical treatment. It still remains unclear how these differences in clinical behavior relate to pathological type. METHODS: A retrospective diagnostic, surgical, and histopathological study of 86 consecutive patients with various pathological types of vertebral hemangioma was performed to establish clinicopathological correlates. RESULTS: The study confirmed that differences exist in clinical course, appearance on imaging, and outcomes in pathological types of hemangiomas. Based on these findings the authors attempted to identify signs characterizing each group. CONCLUSIONS: Differences in clinical history and radiological features exist among pathological types of vertebral hemangiomas. These differences cannot precisely predict the type pathology before histologic examination, but do help us to understand the natural history of such lesions more fully. PMID- 9870816 TI - The jugular foramen: a comparative radioanatomic study. AB - BACKGROUND: Advances in microsurgical techniques made possible the removal of advanced jugular foramen (JF) lesions, which once had been accepted as unoperable. However, successful surgery requires detailed knowledge of the JF anatomy. METHODS: Sixteen jugular foramina in eight formalin-preserved adult cadavers were scanned with axial and coronal high resolution computed tomography (HRCT) prior to dissection. After craniectomy and removal of brain tissue, the relationships of the neurovascular structures in the JF were determined by drilling the temporal bones from superior to inferior on planes parallel to the skull base. RESULTS: No bony partition of the JF was observed. A dural band consistently divided the JF into two parts. Anterior to it was the glossopharyngeal nerve (IX) while the vagus (X) and accessory (XI) nerves were located posteriorly. There was a notch in which the IX nerve entered the JF. It was also identified on the CT scans and defined as the glossopharyngeal recess. The IX nerve made a genu within the JF in all specimens. Then, it ran inferiorly through a bony canal in three specimens (18.75%), and through an incomplete bony canal in two (12.5%), which were also defined on the CT images. The inferior petrosal sinus ran through a sulcus anteromedial to the glossopharyngeal recess. The posterior meningeal artery was found to be located between the X and XI nerves within the JF. CONCLUSIONS: This study revealed a complex and highly variable pattern of the relationships of the neurovascular structures in the JF, and their HRCT images correlated well with the anatomic microdissections. PMID- 9870815 TI - "En-bloc" vertebrectomy in the mobile lumbar spine. AB - BACKGROUND: Primary tumors of the vertebral bodies have previously been treated with total or subtotal excision in a piecemeal fashion (intralesional excision). Radiation therapy has been used to help control tumor growth. Recurrence rates with an intralesional, piecemeal removal of vertebral tumors have been unacceptably high. This study describes a method to excise a lumbar vertebra "en bloc," and in the process, to perform a marginal (extralesional) resection of a primary tumor of the mobile lumbar spine that allows for a potential surgical cure. METHODS: A combined posterior-anterior procedure allows for an extralesional, marginal resection of the tumor and the involved vertebra. All posterior bony elements, including the pedicles and the adjacent intervertebral discs, are removed via a posterior approach. An anterior, retroperitoneal approach is then used to remove the vertebral body/tumor as a single specimen. The nerve roots at the involved levels are spared and the spine is instrumented and fused both posteriorly and anteriorly. RESULTS: Three patients successfully had combined posterior-anterior resections of lumbar vertebral chordomas. No permanent neurological complications occurred. Overall morbidity of the procedure was acceptable. At 31-month follow-up, no tumor recurrence has been detected. CONCLUSIONS: "En-bloc" resection of a primary vertebral tumor of the lumbar spine is technically demanding, but potentially curative. The alternative approaches intralesional excision, radiation therapy, or a combination-are unable to cure these tumors. Long-term, 10-year follow-up will be necessary to confirm whether this en-bloc approach provides a surgical cure. PMID- 9870817 TI - The supracondylar approach to the jugular tubercle and hypoglossal canal. AB - BACKGROUND: Circumscribed lesions of the hypoglossal canal and of the jugular tubercle still remain a surgical challenge. So far, transpetrosal, transcondylar suboccipital, and extreme lateral approaches have been used to access this region. These surgical procedures bear a high risk for neurological deficits. Therefore, we introduce a new minimally invasive extradural approach to the hypoglossal canal that also allows access to the lateral aspects of the jugular tubercle. METHODS: After a paramedian retromastoid skin incision, a basal suboccipital craniectomy lateral to the foramen magnum toward the jugular tubercle is performed. With this approach the occipital condyle and the lateral osseous circumference of the foramen magnum are preserved. Drilling extradurally, the dorsal parts of the jugular tubercle are removed. The exposure is extended downward to the posterior margins of the hypoglossal canal and laterally to the jugular bulb, enabling a minimally invasive exposure of the hypoglossal canal, the lateral aspects of the jugular tubercle, and medial aspects of the jugular bulb. RESULTS: Using this supracondylar approach, surgical interventions were performed in three patients suffering from a hypoglossal neurinoma, a cholesterol granuloma extending into the jugular tubercle, and a cyst of the hypoglossal canal, respectively. No additional postoperative neurological deficits were seen. CONCLUSIONS: The supracondylar approach seems to be useful to gain access to benign lesions of the hypoglossal canal and of the jugular tubercle to decompress tumors or cysts. In contrast to previously reported techniques this approach has a low risk of morbidity. The surgical field, however, is restricted laterally by the jugular bulb, medially and basally by the residual occipital condyle and dorsally by the dura. Therefore, this approach is useful to remove small lesions or to perform extended biopsies. Radical removal of large tumors seems to be problematic using this approach. PMID- 9870818 TI - Hormone-like effects of fibroblast growth factors (FGFs) in the nervous system. PMID- 9870819 TI - Current surgical treatment for sphenoorbital meningiomas. AB - BACKGROUND: Originally, the authors used a combined transcranial-transmalar approach for removal of sphenoorbital tumors. METHODS: More recently, when computed tomography (CT) scanning became available, surgical management of sphenoorbital meningiomas included resection of the hyperostosis in the pterional region, orbital roof and lateral wall, and middle fossa floor. Thereafter, intradural and intraorbital components are removed, as well as infiltrated dura mater. Finally, a periosteal flap is placed over the dural defect and bone reconstructed with methylmethacrylate. RESULTS: Of 25 patients, there was only one death and all but one showed very good to moderate response to treatment. Four recurrences were observed. CONCLUSIONS: Surgical management cannot be uniform in all cases, but must be adapted to each form of presentation. On the basis of the satisfactory results achieved, we feel justified in advising the technique described above with its current modifications. PMID- 9870820 TI - Ki-67 (clone MIB-1) proliferation index in recurrent glial neoplasms: no prognostic significance. AB - BACKGROUND: To determine if the Ki-67 (MIB-1 clone) proliferative index (PI) has prognostic potential in patients with recurrent astroglial neoplasms. METHODS: We conducted a retrospective review of 27 patients whose initial and recurrent specimens were available. Histopathology was determined according to the World Health Organization classification. Proliferation index was calculated on formalin-fixed tissue using the Ki-67 (MIB-1 clone) antibody. Morphometric data were analyzed in conjunction with clinical data and Cox Proportionate Hazards Analysis, Spearman's correlation coefficient and Mann-Whitney Test. RESULTS: Initial histopathology included 14 glioblastoma multiforme, 7 anaplastic astrocytoma, 3 oligoastrocytoma, and 3 astrocytoma. Recurrent specimens showed changes consistent with treatment. While univariate analysis shows initial histology correlated with survival (p<0.036), PI did not correlate with survival after either initial (p = 0.86) or recurrent (p = 0.46) surgery for any tumor type. PI difference between specimens also did not correlate with survival (p = 0.91). Initial PI did not correlate with recurrent PI either (p = 0.43). CONCLUSIONS: Ki-67 PI does not confer additional prognostic information for patients with recurrent astroglial neoplasms. One possible explanation for this observation is that treatment may alter the PI independent of its effect on tumor growth. PMID- 9870822 TI - Cerebral aneurysm associated with von Recklinghausen's neurofibromatosis: a case report. AB - BACKGROUND: von Recklinghausen's neurofibromatosis is a hereditary disease that may affect any organ or system of the body primarily or secondarily, including the vascular system. Among the rare cerebrovascular abnormalities, the most common is stenosis or occlusion of the cerebral artery. Intracranial aneurysms are uncommon. CASE DESCRIPTION: A case of an intracranial cerebral aneurysm associated with von Recklinghausen's neurofibromatosis is reported. A 55-year-old woman presented with a history of intermittent headache for 2 months and right oculomotor nerve palsy for 1 month. Widespread cutaneous neurofibromas and angiomas were found over her trunk and limbs with prominent cafe-au-lait spots. X ray showed that her left lung was compressed by a large mass in the left chest with rib defects and lateral spinal curvature. Right internal carotid angiography revealed a saccular aneurysm between C1 and C2. Craniotomy to clip the aneurysm could not be performed because the mass in her chest made intubation for general anaesthesia almost impossible. CONCLUSION: The clinical features of this case are discussed together with a review of 15 similar cases in the literature. There are different theories about this disorder. We agree that the malformations are derived not only from ectodermal, but also from mesodermal pathology. In terms of our case, we consider the progression of this disease to be slow. PMID- 9870821 TI - Recurrence in meningeal hemangiopericytomas. AB - BACKGROUND: Meningeal hemangiopericytomas are more aggressive than typical meningiomas, with a high rate of recurrence and distant metastases. The question of whether a correlation exists between prognosis and histologic features remains controversial. CASE DESCRIPTION: We report two cases of recurrent meningeal hemangiopericytomas. Although local growth control of the tumor was obtained by tumor removal and irradiation in a 38-year-old male patient (Case 1) with a recurrent tentorial tumor, the tumor disseminated and metastasized extracranially within a short period after treatment, leading to rapid deterioration. Another 38 year-old female patient (Case 2) with a recurrent orbital tumor had a favorable outcome after tumor removal. The Ki-67 proliferative index using the MIB-1 monoclonal antibody increased as the tumor recurred in Case 1 (2.5%, 7.9%, and 15.7%), but did not change between primary and recurrent tumors of Case 2 (4.2%, 3.1%). Immunostaining for p53 protein in Case 1 was negative at the first resection, and became positive at the second and third resections, whereas in Case 2, it was negative in both the primary and recurrent tumors. CONCLUSIONS: Our results suggest that p53 protein accumulation with a high proliferative potential is a useful marker to estimate malignant progression in meningeal hemangiopericytomas. PMID- 9870823 TI - A simple method of scalp localization using multiplanar reconstruction of MR images. AB - BACKGROUND: Image-based scalp localization methods currently used are complex and not standardized. The authors have developed a simple yet accurate method for craniotomy localization using multiplanar reconstruction (MPR) algorithms. METHODS: In this method, the goal is to localize a projected point (defined as T) of the center of the lesion on the scalp. An oblique coronal plane is reformatted using a patient's magnetic resonance (MR) images with MPR algorithms to include both the center of the lesion and bilateral external auditory meati. Then the distance between T and the ipsilateral external auditory meatus or sagittal suture (defined as S) is measured along the scalp contour in the plane. The distance between the bregma and S is also measured. These distances are used for scalp localization, using a tape measure in the operating room. RESULTS: We have had successful scalp localization in six clinical cases. It took about 3 min to measure each distance on the MR console. CONCLUSION: This method for craniotomy planning using MPR algorithms is simple and sufficiently accurate. PMID- 9870825 TI - 1998: a year of global changes with profound effects on neurosurgeons: what you can do in this changing world. PMID- 9870824 TI - Slowly progressive bilateral enophthalmos from metastatic breast carcinoma. PMID- 9870826 TI - Pediatric cranial golf injuries--an emerging contemporary phenomenon? PMID- 9870827 TI - Influencing politicians. PMID- 9870828 TI - Methylene blue. PMID- 9870829 TI - Increasing academic internal medicine's investment in female faculty. PMID- 9870830 TI - Effects of multidisciplinary case management in patients with chronic renal insufficiency. AB - PURPOSE: Though case management has been recommended to improve the outcomes of patients with costly or morbid conditions, it has seldom been studied in controlled trials. We performed a randomized, controlled clinical trial of an intensive, multidisciplinary case management program for patients with chronic renal insufficiency and followed patients for 5 years. PATIENTS AND METHODS: We enrolled 437 primary-care patients (73% of those eligible) with chronic renal insufficiency (estimated creatinine clearance consistently < 50 mL/min with the last serum creatinine level > 1.4 mg/dL) who were attending an urban academic general internal medicine practice. The intensive case management, administered during the first 2 years after enrollment, consisted of mandatory repeated consultations in a nephrology case management clinic staffed by two nephrologists, a renal nurse, a renal dietitian, and a social worker. Control patients received usual care. Primary outcome measurements included serum creatinine level, estimated creatinine clearance, health services use, and mortality in the 5 years after enrollment. Secondary measures included use of renal sparing and potentially nephrotoxic drugs. RESULTS: There were no differences in renal function, health services use, or mortality in the first, second, or third through fifth years after enrollment. There were significantly more outpatient visits among intervention patients, mainly because of the added visits to the nephrology case management clinic. There were also no significant differences in the use of renal sparing or selected potentially nephrotoxic drugs. The annual direct costs of the intervention were $89,355 ($484 per intervention patient). CONCLUSION: This intensive, multidisciplinary case management intervention had no effect on the outcomes of care among primary-care patients with established chronic renal insufficiency. Such expensive and intrusive interventions, despite representing state-of-the-art care, should be tested prospectively before being widely introduced into practice. PMID- 9870831 TI - Incidence and predictors of major hemorrhagic complications from thrombolytic therapy in patients with massive pulmonary embolism. AB - PURPOSE: The risk factors for bleeding in patients receiving recombinant tissue type plasminogen activator for massive pulmonary embolism are not known. PATIENTS AND METHODS: The hospital records of 132 consecutive patients who received recombinant tissue-type plasminogen activator for massive pulmonary embolism were retrospectively reviewed. Bleeding was estimated by using the bleeding severity index, a method previously validated in patients receiving anticoagulants. Multivariate stepwise logistic regression was used to identify independent risk factors for bleeding. Four other definitions of bleeding in large pulmonary embolism thrombolytic trials were also used, and the agreement among these criteria was assessed. RESULTS: According to the bleeding severity index, 33 patients (25%) had one or more major bleeding complications. Hemorrhage at the venous puncture site for angiography was the most frequent complication (15 patients, 11%). Major bleeding at the catheterization site was more common at the femoral site (14 of 63 patients = 22%) than at the brachial site (1 of 63 patients = 2%; P = 0.0004). The use of the five different bleeding definitions resulted in a variation in the major bleeding rate from 3% to 43%. The kappa coefficient varied from 0.07 to 0.84, indicating poor agreement between most of the classifications. CONCLUSION: The use of the femoral vein for pulmonary angiography was the only variable significantly associated with major bleeding. Most of the differences observed in the pulmonary embolism thrombolytic trials are likely related to the differences in the definition of bleeding rather than to the thrombolytic regimen. PMID- 9870832 TI - A randomized comparison of fluconazole with amphotericin B as empiric anti-fungal agents in cancer patients with prolonged fever and neutropenia. AB - PURPOSE: Several studies have documented the efficacy of amphotericin B as empiric antifungal therapy in cancer patients with prolonged fever and neutropenia. Amphotericin, however, is a toxic drug. Fluconazole has broad spectrum antifungal activity with an excellent safety profile. Although prophylactic use of fluconazole is widespread, its efficacy as an empiric antifungal agent has not been extensively investigated. PATIENTS AND METHODS: We randomly assigned 106 patients with absolute neutropenia (< or = 500 cells microL) and persistent fever of undetermined origin (> 38 degrees C) despite 1 week of broad-spectrum antibiotic therapy to receive either fluconazole 400 mg orally daily or amphotericin B 0.5 mg/kg/day. Patients with obvious invasive fungal infections were excluded, as were those with abnormal renal or hepatic function. Success was defined as defervescence with the initially assigned antifungal regimen without development of clinically evident invasive fungal infection. RESULTS: Six patients were excluded from the analysis, mostly because they did not have severe neutropenia. Forty-eight patients received amphotericin B, and 52 received fluconazole. Baseline clinical characteristics and laboratory parameters as well as duration of neutropenia (7.7 versus 6.9 days), duration of fever (7.8 versus 8.1 days), and duration of hospitalization (10.4 versus 8.3 days) were similar between those receiving amphotericin and fluconazole. Treatment success rates and mortality rates were similar in the two groups: 22 (46%) patients in the amphotericin group and 29 (56%) patients in the fluconazole group responded successfully to therapy (P = 0.3), whereas 16 (33%) patients in the amphotericin group and 14 (27%) patients in the fluconazole group died during hospitalization (P = 0.5). Adverse events such as chills and fever (4 versus 1), bronchospasm (2 versus none), severe hypokalemia (25 versus 12) and nephrotoxicity (9 versus 3) were more frequently observed in patients receiving amphotericin. Adverse prognostic factors included prolonged duration of neutropenia and pneumonia. CONCLUSIONS: These results suggest that fluconazole is an equally effective but less toxic alternative to amphotericin B as empiric antifungal therapy in cancer patients with prolonged fever and neutropenia. PMID- 9870833 TI - Clinical and immunological study of 7 patients with minocycline-induced autoimmune phenomena. AB - PURPOSE: Prolonged treatment with minocycline for acne vulgaris has been associated with the development of arthralgia, arthritis, and other autoimmune phenomena. We characterized the clinical, laboratory, and immunological profiles of seven patients with this syndrome. SUBJECTS AND METHODS: Clinically the patients were studied with special emphasis on prior minocycline treatment, presenting symptoms, physical findings, course, and outcome. Laboratory tests included fluorescent antinuclear and antineutrophil cytoplasmic (ANCA) antibodies, as well as antibodies to myeloperoxidase, bactericidal permeability increasing protein, elastase, cathepsin G, lactoferrin, cardiolipin, and histone. RESULTS: All 7 patients presented with polyarthritis or arthralgia, morning stiffness, and fever after 6 to 36 months of minocycline treatment. The skin was involved in five patients (three with livedo reticularis and two with subcutaneous nodules). Two patients had chronic active hepatitis. Increased titers of perinuclear ANCA (p-ANCA) were detected in all seven patients; five patients had fluorescent antinuclear antibodies, two had antihistone autoantibodies and one had anticardiolipin antibodies. Antigenic characterization of p-ANCA disclosed antibodies to bactericidal permeability increasing protein in one patient, to elastase in three patients, and to cathepsin G in five patients. Symptoms resolved in five patients upon discontinuation of minocycline; the other two patients were treated with corticosteroids and also achieved remissions. CONCLUSION: Minocycline-induced autoimmune syndrome is characterized by reversible polyarthralgia or arthritis, morning stiffness, fever, frequent skin involvement, occasional chronic active hepatitis, and increased titers of p-ANCA with various minor p-ANCA-related antigens. PMID- 9870834 TI - Serum fructosamine level and the risk of hip fracture in elderly women: a case cohort study within the study of osteoporotic fractures. AB - PURPOSE: While a high serum fructosamine level may be an indicator of undiagnosed diabetes, a low level may be indicative of poor nutrition or frailty. As malnutrition is a risk factor for osteoporosis, low serum fructosamine levels may be associated with an increased risk of osteoporotic fracture. We examined the association between serum fructosamine levels and the risk of subsequent hip and vertebral fracture. SUBJECTS AND METHODS: We performed a case-cohort study within the Study of Osteoporotic Fractures. Subjects were elderly, ambulatory, community dwelling, Caucasian, women. Fructosamine levels were measured in baseline serum. Incident vertebral fractures were identified by comparing baseline spinal radiographs to those obtained an average of 3.5 years later. Hip fractures were confirmed by radiograph. We randomly selected 101 women who suffered a hip fracture, 100 women who developed a vertebral fracture, and 276 controls. We compared fructosamine levels in women with subsequent osteoporotic fractures to controls. All analyses were adjusted for age, weight, and use of estrogens. RESULTS: Women with fructosamine levels in the lowest decile (< or = 223 micromol/L) had a three-fold increase in the risk of hip fracture (95% confidence interval 1.4-6.4), compared with all other women. Adjustment for markers of frailty, including smoking, functional status, and serum albumin levels, reduced the strength of this association. No clear association was observed between serum fructosamine level and the risk of vertebral fracture. CONCLUSION: Low serum fructosamine levels, which likely reflect frailty or malnutrition, may be a useful clinical tool to identify women at risk for hip fracture. PMID- 9870835 TI - Effects of aspirin treatment on survival in non-insulin-dependent diabetic patients with coronary artery disease. Israeli Bezafibrate Infarction Prevention Study Group. AB - PURPOSE: The benefit of aspirin treatment among diabetic patients with chronic coronary artery disease is not well established. The purpose of this study was to assess the effect of aspirin on cardiac and total mortality in a large cohort of diabetic patients with established coronary artery disease and to compare it with the effect of aspirin in nondiabetic counterparts. PATIENTS AND METHODS: In this observational study among patients screened for participation in the Bezafibrate Infarction Prevention Study, the effects of aspirin treatment in 2,368 non insulin-dependent diabetic patients with coronary artery disease were compared to those in 8,586 nondiabetic patients. Hazard ratios (HR) and 95% confidence intervals (CI) were estimated with proportional hazards models. RESULTS: Fifty two percent of diabetic patients and 56% of nondiabetic patients reported aspirin therapy. After 5.1 +/- 1.3 (mean +/- SD) years of follow-up, the absolute benefit per 100 patients treated with aspirin was greater in diabetic patients than in nondiabetic patients (cardiac mortality benefit: 5.0 versus 2.1, and all-cause mortality benefit: 7.8 versus 4.1). Overall cardiac mortality among diabetic patients treated with aspirin was 10.9% versus 15.9% in the nonaspirin group (P < 0.001), and all-cause mortality was 18.4% and 26.2% (P < 0.001). After adjustment for possible confounders, treatment with aspirin was an independent predictor of reduced overall cardiac (HR = 0.8; 95% CI: 0.6-1.0) and all-cause mortality (HR = 0.8; 95% CI: 0.7-0.9) among diabetic patients, similar to those in nondiabetic patients. CONCLUSION: Treatment with aspirin was associated with a significant reduction in cardiac and total mortality among non-insulin-dependent diabetic patients with coronary artery disease. The absolute benefit of aspirin was greater in diabetic patients than in those without diabetes. PMID- 9870836 TI - Effect of diabetes mellitus on the presentation and triage of patients with acute chest pain without known coronary artery disease. AB - PURPOSE: Patients with diabetes and acute chest pain may be admitted to hospitals more frequently than patients without diabetes because physicians suspect atypical presentations for ischemic heart disease. This study aimed to determine whether the presentation of acute myocardial infarction and risk for major cardiac complications differs among patients without known coronary artery disease who do or do not have diabetes. PATIENTS AND METHODS: Data from an emergency department of an urban teaching hospital on the medical histories, physical examinations, and electrocardiograms of 2,694 subjects with acute chest pain and without known coronary artery disease were prospectively recorded. RESULTS: Diabetes was present in 301 (11%) patients. Compared with patients without diabetes, patients with diabetes were more likely to be < or = 60 years old (51% versus 20%) and to have a history of hypertension (70% versus 35%) or high blood cholesterol (35% versus 19%). A discharge diagnosis of acute myocardial infarction was made in 25 diabetic (8%) and in 148 nondiabetic (6%; P = 0.16) patients. A major cardiac complication occurred in two patients with diabetes (0.7%) and in 20 patients without diabetes (0.8%; P = 1.0). Patients with and without diabetes who had atypical chest pain complaints had similar rates of myocardial infarction (3% and 4%, respectively; P = 0.6). Patients with diabetes were more likely to be hospitalized (67% versus 47%; P = 0.001) both before and after adjusting for clinical and electrocardiographic data. CONCLUSIONS: For patients with acute chest pain without a prior history of coronary artery disease, diabetes was not associated with a higher rate of acute myocardial infarction or complications. However, diabetes was associated with a higher rate of hospitalization in this population, suggesting that physicians have a lower threshold for admission to the hospital of patients with diabetes. PMID- 9870837 TI - Hospital admissions through the emergency department: does insurance status matter? AB - PURPOSE: To assess the effect of insurance status on the probability of admission and subsequent health status of patients presenting to emergency departments. SUBJECTS AND METHODS: We performed a prospective cohort study of patients with common medical problems at five urban, academic hospital emergency departments in Boston and Cambridge, Massachusetts. The outcome measure for the study was admission to the hospital from the emergency department and functional health status at baseline and follow-up. RESULTS: During a 1-month period, 2,562 patients younger than 65 years of age presented with either abdominal pain (52%), chest pain (19%) or shortness of breath (29%). Of the 1,368 patients eligible for questionnaire, 1,162 (85%) completed baseline questionnaires, and of these, 964 (83%) completed telephone follow-up interviews 10 days later. Fifteen percent of patients were uninsured and 34% were admitted to the hospital from the emergency department. Uninsured patients were significantly less likely than insured patients to be admitted, both when adjusting for urgency, chief complaint, age, gender and hospital (odds ratio = 0.5, 95% confidence interval 0.3 to 0.7), and when additionally adjusting for comorbid conditions, lack of a regular physician, income, employment status, education and race (odds ratio = 0.4, 95% confidence interval 0.2 to 0.8). However, there were no differences in adjusted functional health status between admitted and nonadmitted patients by insurance status, either at baseline or at 10-day follow-up. CONCLUSIONS: Uninsured patients with one of three common chief complaints appear to be less frequently admitted to the hospital than are insured patients, although health status does not appear to be affected. Whether these results reflect underutilization among uninsured patients or overutilization among insured patients remains to be determined. PMID- 9870838 TI - Effective management of gout: an analogy. PMID- 9870839 TI - Risk reduction in the intensive care unit. AB - Many potentially preventable complications occur in patients who receive intensive care. We have reviewed the epidemiology of three important complications (venous thromboembolism, stress-related upper gastrointestinal bleeding, and vascular catheter-related infection) and evaluated common preventive treatments to provide evidence-based recommendations for prevention. We used English language articles located by MEDLINE or cross-citation, giving preference to articles published in the last 10 years, meta-analyses, and clinical trials that were randomized, double-blinded, and used intention-to-treat analysis. We recommend prophylaxis against venous thromboembolism in most patients, whereas those without respiratory failure or coagulopathy may not require prophylaxis against stress-related upper gastrointestinal hemorrhage. Chlorhexidine gluconate is the preferred antiseptic for disinfecting the skin prior to and during intravascular catheterization. Central venous catheters impregnated with antibacterial or antiseptic agents should be considered in patients at high risk for vascular catheter-related infection. Finally, central venous, pulmonary arterial, and systemic arterial catheters should be changed only when clinically indicated. PMID- 9870840 TI - Thyroid disease mediated by molecular defects in cell surface and nuclear receptors. AB - The proposed mechanisms of RTH are not mutually exclusive. In fact, there is considerable experimental evidence that many if not all of these complex receptor interactions with elements of the transcriptional unit are involved in RTH. Several aspects of RTH remain unclear, in particular on a clinical level. We still do not completely understand the seeming paradox of a tight distribution of receptor mutations and wide variability in phenotypic presentation. The discovery that many of the RTH receptors have defects in corepressor interaction makes it tempting to speculate that the variability in RTH phenotype within kindreds is secondary to differences in corepressor expression. These issues may be better understood as research further proceeds into cofactors and their control of transcription. We also need better tools to determine thyroid status at a peripheral level. Basal metabolic rate, serum measurement of thyroid-responsive gene products, echocardiographic techniques, and other clinical measures have for the most part been unhelpful in determining thyroid status of specific organ systems. Consequently, therapeutic interventions for RTH are directed toward normalizing biochemical indices of thyroid homeostasis, without really knowing whether these efforts correct imbalances within crucial tissues. These studies, and the more widespread investigation of hormone receptor action in general, are moving at a breathtaking pace, and there is a keen interest in applying these principals to understanding the pathophysiologic mechanism of a variety of diseases. PMID- 9870841 TI - Reciprocal changes in progression of gastric mucosa-associated lymphoid tissue lymphoma and pancreatic somatostatinoma. PMID- 9870842 TI - Enhancing progressive multifocal leukoencephalopathy: an indicator of improved immune status? PMID- 9870843 TI - Evaluating the consequences of multidisciplinary case management for patients with chronic renal failure. PMID- 9870844 TI - Living with gout: preventing the fire. PMID- 9870845 TI - Systematic reviews and evidence-based critical care medicine: a step in the right direction. PMID- 9870846 TI - Systemic capillary leak syndrome. PMID- 9870847 TI - Effects of gonadotropin-releasing hormone agonists on uterine leiomyomas. PMID- 9870848 TI - Testing for resistance to activated protein C factor V R506Q. PMID- 9870849 TI - Rhabdomyosarcoma mimicking acute leukemia. PMID- 9870850 TI - Benign breast diseases and premalignant breast disease. PMID- 9870851 TI - Extension of useful reagent shelf life beyond manufacturers' recommendations. Cell Markers Committee of the College of American Pathologists. AB - OBJECTIVE: The College of American Pathologists Cell Markers Committee designed a study to evaluate the use of immunohistochemistry primary antibodies beyond manufacturers' recommended dates. METHODS: Pathologists were asked to save aliquots of primary antibodies during mid-1997 so that by spring 1998 the reagents would be "outdated" according to manufacturers' recommendations. Three tumors were immunostained both in mid-1997 and early 1998 (using outdated reagents in 1998). Two hundred twenty-one laboratories participated. PATIENT SAMPLES: Immunostained materials consisted of an angiomyolipoma immunostained for muscle-specific actin and HMB-45, a melanoma immunostained for S100 protein and HMB-45, and a large cell lymphoma immunostained for common leukocyte antigen and HMB-45. Blocks from the same tumor were used in each instance. MAIN OUTCOME MEASURE: We compared the immunostaining results as a percentage of laboratories indicating a positive or negative immunohistochemical result between the 1997 and 1998 time points. RESULTS: Only minor differences were identified for the 221 reporting laboratories in 1998 as compared with those in 1997. CONCLUSIONS: The data suggest review of the Health Care Financing Administration's ruling on extending the useful reagent shelf life beyond manufacturers recommendations. Similar studies using more inherently quantitative methodology are suggested. PMID- 9870852 TI - Benign breast changes and the risk for subsequent breast cancer: an update of the 1985 consensus statement. Cancer Committee of the College of American Pathologists. AB - The Cancer Committee of the College of American Pathologists has prepared an update of the consensus statement on premalignant breast lesions and breast cancer risk that was originally published in the Archives of Pathology & Laboratory Medicine in 1986. The objective of this publication is to better define the relative breast cancer risk associated with specific histologic abnormalities by incorporating data derived from recent case-control studies. Explanatory notes are used to document and explain specific risk classifications. In addition to refining the degree of risk associated with individual lesions, such as fibroadenoma and atypical hyperplasia, this update includes a discussion of age-specific breast cancer risk and provides examples that can be used when counseling patients. PMID- 9870853 TI - Practice guideline for forensic pathology. Members of the Forensic Pathology Committee, College of American Pathologists. AB - The guideline for forensic pathology was prepared by the Forensic Pathology Committee of the College of American Pathologists. The definitional criteria for forensic pathology included in this guideline have been approved both by the House of Delegates and the Board of Governors of the College of American Pathologists. The guideline presents an overview of forensic pathology and an approach to the forensic autopsy and medicolegal death investigation. Emphasis is placed on the role of forensic pathology in maintaining public health, welfare, and safety. The guideline is intended to serve as an educational tool, and its use should be determined by the individual circumstances and settings of specific cases. PMID- 9870854 TI - Providing human tissues for research: how to establish a program. AB - The Cooperative Human Tissue Network is a group composed of cooperating academic institutions that supply human tissues to researchers studying a wide range of neoplastic and other diseases. The experience of the Cooperative Human Tissue Network in establishing methods of prospective tissue collection and in developing tumor banks is discussed to aid institutions in establishing tissue resources for their local investigators, who may wish to use human tissues in current or future research projects. The advantages to pathology departments and to associated medical institutions of establishing an organized tissue resource include ensuring proper institutional review board approval of research projects using human tissues, protecting diagnostic specimens, creating new opportunities for extramural research, increasing the speed of diagnostic specimen transport to surgical pathology, and providing educational and research opportunities for pathologists and pathology residents. Methods of tissue collection, processing, storage, data collection, and supply are outlined. Also, resources necessary to begin organized tissue collection, including personnel, space, equipment, and supplies, are discussed. PMID- 9870855 TI - Perianal Paget's disease: distinguishing primary and secondary lesions using immunohistochemical studies including gross cystic disease fluid protein-15 and cytokeratin 20 expression. AB - BACKGROUND: Extramammary Paget's disease most commonly occurs on the female external genitalia and rarely occurs in the perianal region and male external genitalia. We present the clinical and pathologic features of 5 cases of perianal Paget's disease and review the literature. METHODS: Clinical and pathologic data were recorded for 5 cases of perianal Paget's disease. Cases were studied retrospectively with special stains, including periodic acid-Schiff, mucicarmine, Alcian blue, carcinoembryonic antigen, S100 protein, pan-keratin, gross cystic disease fluid protein-15 (GCDFP-15), lysozyme, CD15 (Leu-M1), cytokeratin 7 (CK7), and cytokeratin 20 (CK20). RESULTS: Three (60%) of 5 patients had concurrent rectal adenocarcinomas. All cases reacted positively for pankeratin, although the intensity and distribution of staining varied. Both cases not associated with an underlying carcinoma showed strong GCDFP-15 and CK7 expression and an absence of CK20 expression. The 3 cases associated with an underlying malignancy demonstrated CK7 and CK20 expression and an absence of GCDFP-15 expression. All cases were negative for lysozyme and CD15 (Leu-M1). CONCLUSIONS: The 5 cases reported herein demonstrate that perianal Paget's disease is a heterogeneous entity. The high frequency of associated underlying malignancies and resultant poor clinical outcomes highlight the importance of an aggressive search for a second malignancy. In some cases, perianal Paget's disease merely represents a cutaneous manifestation of an underlying rectal adenocarcinoma and demonstrates a CK7+/CK20+/GCDFP-15-/lysozyme-/Leu-M1- immunophenotype and signet ring Paget's cells. Other cases represent primary adenocarcinomas of the skin, which are associated with a CK7+/CK20-/GCDFP-15+/lysozyme /Leu-M1- immunophenotype and an excellent prognosis if adequately resected. Immunohistochemical studies, particularly CK20 and GCDFP-15, are useful adjuncts in distinguishing primary and secondary perianal Paget's disease. PMID- 9870856 TI - Anaplastic pleomorphic xanthoastrocytoma. AB - Well-documented cases of malignant degeneration in pleomorphic xanthoastrocytoma and of anaplastic pleomorphic xanthoastrocytoma are rare in the literature. We report 2 cases of pleomorphic xanthoastrocytoma, 1 of which demonstrated clear evidence of malignant degeneration in the absence of prior radiation therapy over an 18-year period. Both anaplastic tumors were characterized by foci of necrosis and increased mitotic activity (3 and 2 mitotic figures/10 high-power fields). Both tumors demonstrated focal positive staining for glial fibrillary acidic protein and showed marked reticulin deposition. An MIB-1 labeling index (marker of cell proliferation) in the initial low-grade-appearing tumor in case 1 was 0.1%. The recurrent tumor in case 1 had an MIB-1 labeling index of 4.9%, and the anaplastic tumor in case 2 had an MIB-1 labeling index of 5.4%. Significant cyclin D1 immunoreactivity was not observed in either anaplastic tumor. Two percent to 3% of tumor cells stained positive with p53 protein antibody in the recurrent anaplastic tumor in case 1. Although histology may not reliably predict aggressive behavior in pleomorphic xanthoastrocytomas, the presence of increased mitosis, necrosis, and increased cell proliferation labeling indices may be indicative of a higher grade tumor. PMID- 9870857 TI - Possible relation of osteopontin to development of psammoma bodies in human papillary thyroid cancer. AB - BACKGROUND: Psammoma bodies are often observed in papillary carcinomas of the thyroid. Recently we reported that osteopontin (OPN) appears to play an important role in the development of psammoma bodies in meningiomas. Because the morphology and components of psammoma bodies in papillary carcinoma of the thyroid are similar to those of psammoma bodies in meningioma, we examined whether OPN was related to the psammoma bodies in papillary carcinoma of the thyroid. METHODS: Expression of OPN mRNA was examined by Northern blotting and in situ hybridization. Immunohistochemistry was performed to localize the deposition of OPN protein and to identify the type of OPN mRNA-expressing cells. RESULTS: The expression of OPN mRNA was detected in papillary thyroid carcinomas but not in normal thyroid tissue. Osteopontin mRNA-expressing cells were present around the psammoma bodies, and the localization of OPN protein was consistent with that of psammoma bodies. The OPN mRNA-expressing cells were identified as CD68-positive macrophages. CONCLUSION: Osteopontin produced by macrophages may play a significant role in the development of psammoma bodies in papillary carcinoma of the thyroid. PMID- 9870858 TI - Placental lesions associated with neurologic impairment and cerebral palsy in very low-birth-weight infants. AB - OBJECTIVE: Systematic placental examination has the potential to shed light on poorly understood antenatal processes that may increase the risk of neurologic impairment and cerebral palsy. DESIGN: Using data from a retrospective case control study, we analyzed placentas from 60 inborn, singleton, very low-birth weight (<1.5 kg) infants delivered between 1983 and 1991 who had subsequent neurologic impairment at 20 months corrected age (42 with cerebral palsy and 18 with other neurologic abnormalities) and 59 control infants of comparable gestational age, birth weight, sex, and race. Three a priori hypotheses based on previous studies were that neurologic impairment would be increased with fetal vascular lesions with or without coexisting chorioamnionitis, decreased with chronic maternal vascular underperfusion, and increased when multiple placental abnormalities were seen in the same case. RESULTS AND CONCLUSIONS: We found 2 types of fetal placental vascular lesions to be associated with neurologic impairment, namely, recent nonocclusive thrombi of chorionic plate vessels (P < .04) and severe villous edema (P < .01). Chorionic plate thrombi were seen only with chorioamnionitis and accounted for the increased risk of neurologic impairment seen with chorioamnionitis. Maternal vascular lesions showed a biphasic relation to neurologic impairment in the subgroup of patients without chorioamnionitis. Mild lesions were increased in controls (inadequate vascular remodeling, P=.03, and accelerated maturation, P=.004). A more severe lesion, multiple villous infarcts, although not reaching significance, was increased in the neurologically impaired cases. Finally, in a test of 9 selected placental lesions, cases with cerebral palsy were more likely to have 2 or more lesions (P < .0001) and were less likely to have no lesions (P < .04) than control infants. PMID- 9870859 TI - Congenital malignant rhabdoid tumor presenting as a cutaneous nodule: report of 2 cases with review of the literature. AB - OBJECTIVE: The congenital occurrence of malignant rhabdoid tumor (MRT) is rare. Initial presentation of congenital MRT as a skin tumor is even more rare. We report 2 cases in which a cutaneous nodule appeared prior to the discovery of the primary tumor. METHODS: Histologic, ultrastructural, and immunohistochemical studies were performed on skin nodules from 2 neonates. Other congenital examples of MRT in the literature were reviewed. RESULTS: Both neonates were male and both were fullterm. Skin nodules were noted at birth. The tumors were located on the right neck in case 1 and on the right back in case 2. The primary tumors were later found in the kidney and the right chest wall, respectively. The skin biopsy of the first case had been interpreted as undifferentiated sarcoma, and MRT was diagnosed after the renal tumor was examined. In the second case, a correct diagnosis was promptly made based on the skin biopsy owing to experience with the first case. CONCLUSIONS: Congenital MRT is very uncommon and may present as a skin tumor at birth. Since MRT exhibits various histologic patterns, the diagnosis should be made by a complete pathologic study with proper clinical correlation. To our knowledge, the unique mode of manifestation in these 2 cases is described for the first time. We suggest that MRT should be considered when making the differential diagnosis in such a clinical setting. PMID- 9870860 TI - Lack of asbestos contamination of paraffin. AB - OBJECTIVE: To investigate possible asbestos contamination of paraffin and migration by asbestos fibers during the tissue-embedding process. DESIGN: Three sample categories were included in the study: (1) commercially available paraffin samples; (2) procedural control samples, which were prepared by processing the paraffin through the use of standard solvents and instruments; and (3) samples taken from areas adjacent to embedded tissue and evaluated for migration of asbestos from the tissue into the surrounding paraffin. The analysis of collected material from all samples was performed with analytical transmission electron microscopy. RESULTS: Only one extremely small tremolite fiber was found in any of the commercially available samples of paraffin. No asbestos fibers were found either in the procedural control samples or in the samples taken adjacent to the embedded lung tissue. CONCLUSIONS: First, it was extremely unlikely that any of the commercial paraffin samples would have skewed data due to embedded tissue. Second, the processing and instrumentation was not found to contribute asbestos material to the paraffin during the preparations. Finally, embedded tissue that contained high numbers of fibers, both uncoated fibers and asbestos bodies, did not contribute asbestos to the adjacent paraffin. PMID- 9870861 TI - Fontana-Masson--stained tissue from culture-proven mycoses. AB - BACKGROUND: The Fontana-Masson stain can be used to detect melanin of dematiaceous fungi such as Curvularia lunata and Bipolaris hawaiiensis in tissue. This stain is often used to identify fungi that may not have an evident brown color in hematoxylin-eosin-stained sections. However, it is not clear whether hyphae of nondematiaceous fungi can be stained with the Fontana-Masson procedure. METHODS: Paraffin sections from culture-proven mycoses caused by species of Aspergillus, Candida, Fusarium, Pseudallescheria, Trichosporon, zygomycetes, and dematiaceous fungi were stained by the Fontana-Masson procedure. Extent of stain intensity and its distribution in fungal elements in tissue were evaluated by means of an intensity scale based on Cryptococcus neoformans as a positive control. RESULTS: Many Aspergillus spp, some zygomycetes, and all of the dematiaceous fungi showed strong intensity, whereas other fungi showed either weak intensity or no staining. CONCLUSION: Fungal hyphae other than dematiaceous fungi occasionally can be stained with the Fontana-Masson procedure. The morphology of positively stained hyphae must be carefully evaluated, especially when they are hyaline in hematoxylin-eosin-stained tissue sections. PMID- 9870862 TI - Primary myxoid leiomyoma of the liver. AB - We describe a case of hepatic leiomyoma in a 41-year-old woman who complained of abdominal discomfort due to a right upper quadrant abdominal mass. A computed tomographic scan revealed a large hepatic mass with unusual cystic change. Light and electron microscopic findings and an immunohistochemical study demonstrated diffuse and scattered proliferation of smooth muscle cells in a myxoid matrix. In contrast to previously reported cases of primary leiomyoma of the liver, the present tumor developed in a patient without an immunosuppressive condition. To the best of our knowledge, this is the first reported case of primary "myxoid" leiomyoma of the liver. PMID- 9870863 TI - Neuron-specific enolase-producing epithelioid leiomyosarcoma with gross vascular invasion and intracardiac involvement. AB - We present an unusual case of epithelioid leiomyosarcoma in the right shoulder. A 10-cm tumor eroding the right scapula occurred in a 59-year-old man and grew into the subclavian vein with extension to the right atrium. An elevated serum level of neuron-specific enolase was also detected. Clinical presentation suggested small cell carcinoma of the lung (Pancoast tumor) with intravenous extension. Eventually, the patient died of respiratory insufficiency. Autopsy revealed a primary tumor in the right shoulder, numerous tumor emboli in the pulmonary arteries, and associated hemorrhagic infarction. Histopathologic features revealed an epithelioid leiomyosarcoma. Furthermore, immunohistochemical and immunoblotting analyses of the tumor demonstrated positive reactions for neuron specific enolase. We interpreted this peculiar case to be a neuron-specific enolase-producing epithelioid leiomyosarcoma of the soft tissue associated with gross vascular invasion. PMID- 9870864 TI - Chlamydoconidia formation in zygomycosis due to Rhizopus species. AB - An unusual case of endobronchial zygomycosis, which was caused by Rhizopus species and which disseminated to one kidney, occurred in a 36-year-old, diabetic man. The patient recovered fully following lobectomy, nephrectomy, amphotericin B therapy, and control of diabetes mellitus. An interesting histologic finding was the presence of chlamydoconidia formation within the resected lung lesion. To our knowledge, only one previous culture-proven case of zygomycosis has described chlamydoconidia formation in tissue. PMID- 9870865 TI - The epidemiology of systemic lupus erythematosus in populations of African ancestry: a critical review of the "prevalence gradient hypothesis". PMID- 9870866 TI - Treatment of polyarteritis nodosa and microscopic polyangiitis. PMID- 9870867 TI - Functional subsets of CD4 T cells in rheumatoid synovitis. AB - OBJECTIVE: To identify the functional properties of CD4+ CD28- T cells, which accumulate and clonally expand in patients with rheumatoid arthritis (RA). METHODS: The gene expression of molecules involved in T cell effector functions was compared in CD4+ CD28- and CD4+ CD28+ T cell clones. The expression of differentially up-regulated genes was confirmed by flow cytometry of T cells and by 2-color immunohistochemistry of rheumatoid synovial tissue. Cytotoxicity of CD4+ CD28- T cells was tested by anti-CD3 redirected lysis of Fc receptor positive target cells. RESULTS: CD4+ CD28- T cell clones lacked messenger RNA for the CD40 ligand (CD40L) but transcribed the perforin gene. Perforin was also found in freshly isolated CD4+ CD28- peripheral blood lymphocytes from RA patients. CD4+ CD28-, but not CD4+ CD28+, T cell clones lysed Fc receptor-bearing target cells. CD4+ perforin-positive T cells were present in the synovial tissue, where their frequency correlated with the expansion of the CD4+ CD28- compartment in the periphery. Among tissue-infiltrating CD4+ T cells, only the CD40L-negative subset expressed perforin transcripts. CONCLUSION: Clonally expanded CD4+ CD28- T cells are functionally specialized for killing, while they lack the ability to provide B cell help. Tissue-infiltrating CD4+ T cells can be subdivided phenotypically and functionally into at least 2 distinct subsets based on their expression of perforin and CD40L. Because the expansion of CD4+ CD28- T cells is associated with extraarticular RA, T cell-mediated cytotoxicity may be particularly important in these most severe complications of RA. PMID- 9870868 TI - Blockage of interleukin-6 receptor ameliorates joint disease in murine collagen induced arthritis. AB - OBJECTIVE: To clarify the role of interleukin-6 (IL-6) in the pathogenesis of collagen-induced arthritis (CIA). METHODS: CIA was induced by immunizing twice at a 3-week interval with bovine type II collagen (CII) emulsified with complete adjuvant. Rat anti-mouse IL-6 receptor (anti-IL-6R) monoclonal antibody MR16-1 or isotype-matched control antibody KH-5 was then injected once intraperitoneally. Symptoms of arthritis were evaluated with a visual scoring system, and serum anti CII antibody and IL-6 levels were measured by enzyme-linked immunosorbent assay. In addition, the CII responsiveness of splenic lymphocytes from mice with CIA was examined. RESULTS: In mice with CIA, excess production of IL-6 in sera was observed within 24 hours after the first CII immunization, and then rapidly decreased. Serum IL-6 increased again beginning 14 days after immunization, in conjunction with the onset of arthritis. When MR16-1 was injected immediately after immunization with CII, it inhibited the development of arthritis in a dose dependent manner. Furthermore, MR16-1-treated mice exhibited lower serum levels of IgG anti-CII antibody and reduced responsiveness of lymphocytes to CII. This suppressive effect was observed when MR16-1 was injected on day 0 or 3, but not when injected on day 7 or 14. CONCLUSION: IL-6 produced after CII immunization appears to play an essential role in the immunity to CII, and anti-IL-6R antibody reduces the development of CIA by suppressing IL-6 signal transduction. PMID- 9870869 TI - Identification of a new non-major histocompatibility complex genetic locus on chromosome 2 that controls disease severity in collagen-induced arthritis in rats. AB - OBJECTIVE: To identify novel non-major histocompatibility complex (non-MHC) genetic loci controlling the severity of homologous rat type II collagen-induced arthritis (CIA). METHODS: We conducted a genome-wide scan to identify CIA regulatory quantitative trait loci (QTL) in an F2 cross between DA (CIA highly susceptible) and ACI (CIA resistant) inbred rats immunized with homologous rat type II collagen (RII). These strains share the MHC/RT1av1 haplotype required for susceptibility to RII-induced CIA. RESULTS: F2 females had higher median arthritis scores than did males. Relative resistance in the males was determined by inheriting either a DA or an ACI Y chromosome and was independent of the source of the X chromosome. In addition, a major QTL was localized on chromosome 2 (Cia7, logarithm of odds score 4.6). Cia7 is in a region that shows linkage conservation with chromosomal regions that regulate autoimmune diabetes and experimental autoimmune encephalomyelitis in mice and multiple sclerosis in humans. CONCLUSION: Sex chromosomes and Cia7 play an important role in regulating CIA in response to RII. This rat model should facilitate positional cloning and functional characterization of regulatory genes that may play a role in several forms of autoimmune disease, including rheumatoid arthritis. PMID- 9870870 TI - Transcriptional activation of the alpha1(I) procollagen gene and up-regulation of alpha1(I) and alpha1(III) procollagen messenger RNA in dermal fibroblasts from tight skin 2 mice. AB - OBJECTIVE: To investigate the levels of expression of type I and type III collagen genes in dermal fibroblasts from tight skin 2 (Tsk2) and normal mice and to examine the transcriptional regulation of the alpha1(I) procollagen gene (COL1A1) in these cells. METHODS: Dermal fibroblasts from Tsk2 mice and from normal age- and sex-matched control mice were studied. Steady-state levels of alpha1(I) and alpha1(III) procollagen messenger RNA (mRNA) were evaluated by Northern and dot-blot hybridization analyses. The transcriptional regulation of COL1A1 was examined by transient transfection experiments with deletion constructs containing portions of the COL1A1 promoter ligated to the chloramphenicol acetyltransferase reporter gene. To identify DNA binding proteins that interact with regulatory elements within the COL1A1 promoter, gel mobility shift assays were performed with nuclear extracts prepared from normal and Tsk2 dermal fibroblasts. RESULTS: Synthesis of collagen was almost 100% higher in Tsk2 dermal fibroblasts than in control fibroblasts. Up-regulation of mRNA for 2 extracellular matrix proteins was observed in the Tsk2 dermal fibroblasts compared with the normal cells: the alpha1(I) procollagen mRNA steady-state levels were 50% higher, and those of the alpha1(III) procollagen mRNA 100% higher, in Tsk2 cells. The results of transient transfection experiments with COL1A1 promoter constructs demonstrated that the elevated levels of alpha1(I) collagen mRNA in Tsk2 cells were largely due to increased transcriptional activity of the corresponding gene. Electrophoretic mobility shift assays performed with a probe encompassing a relevant COL1A1 promoter region revealed increased DNA-protein binding activities in nuclear extracts prepared from Tsk2 fibroblasts compared with normal fibroblasts. Competition experiments using consensus Spl and nuclear factor 1 (NF-1) oligonucleotides and supershift experiments using anti-Sp1 and anti-NF-1 antibodies indicated that at least 2 transcription factors, Sp1 and NF-1, or their homologs are involved in the up regulated transcriptional activity of the COL1A1 promoter in Tsk2 fibroblasts. CONCLUSION: Dermal fibroblasts from Tsk2 mice display increased collagen synthesis and up-regulation of alpha1(I) and alpha1(III) procollagen mRNA in vitro. The data also directly demonstrate the transcriptional activation of COL1A1 in dermal fibroblasts from Tsk2 mice and suggest that the transcription factors Sp1 and NF-1 or their homologs play an important role in the upregulated expression of this gene in Tsk2 fibroblasts. These findings are similar to those described for fibroblasts from humans with systemic sclerosis and validate the use of Tsk2 as a model for the study of the connective tissue alterations in this disease. PMID- 9870871 TI - Inhibition of interleukin-1alpha-induced cartilage oligomeric matrix protein degradation in bovine articular cartilage by matrix metalloproteinase inhibitors: potential role for matrix metalloproteinases in the generation of cartilage oligomeric matrix protein fragments in arthritic synovial fluid. AB - OBJECTIVE: To determine whether matrix metalloproteinases (MMPs) degrade cartilage oligomeric matrix protein (COMP) to produce fragments similar to those found in synovial fluid (SF) from patients with arthritis. METHODS: COMP fragments were generated in vitro by treating (a) bovine articular cartilage with interleukin-1alpha (IL-1alpha), (b) purified bovine COMP with MMPs, and (c) articular cartilage with MMPs. The fragments generated in each case were analyzed by Western blot, using an antibody to the C-terminal heptadecapeptide of COMP. RESULTS: IL-1alpha stimulation of cartilage resulted in a fragmentation of COMP, which was inhibited by MMP inhibitors CGS 27023A and BB-94. Isolated, recombinant MMPs rapidly degraded purified COMP, as well as COMP residing in cartilage. Several COMP fragments produced in vitro had similar electrophoretic mobility to those in SF of patients with arthritis. CONCLUSION: MMPs may contribute to the COMP fragments found in vivo. Quantitation of MMP-specific fragments may be useful in the evaluation of MMP inhibitors in patients with arthritis. PMID- 9870872 TI - Parathyroid hormone-related proteins is abundant in osteoarthritic cartilage, and the parathyroid hormone-related protein 1-173 isoform is selectively induced by transforming growth factor beta in articular chondrocytes and suppresses generation of extracellular inorganic pyrophosphate. AB - OBJECTIVE: Parathyroid hormone-related protein (PTHrP) is a major, locally expressed regulator of growth cartilage chondrocyte proliferation, differentiation, synthetic function, and mineralization. Because mechanisms that limit cartilage chondrocytes from maturing and mineralizing are diminished in osteoarthritis (OA), we studied PTHrP expression by articular chondrocytes. METHODS: PTHrP was studied in normal knee cartilage samples and cultured articular chondrocytes, and in cartilage specimens from knees with advanced OA, obtained at the time of joint replacement. RESULTS: PTHrP was more abundant in OA than in normal human knee articular cartilage. Both demonstrated PTH/PTHrP receptor expression. PTHrP 1-173, one of three alternatively spliced PTHrP isoforms, was exclusively expressed and induced by transforming growth factor beta in cultured chondrocytes. Chondrocytes mainly used the GC-rich P2 alternative promoter to express PTHrP messenger RNA. Inhibition by PTHrP 1-173, but not by PTHrP 1-146 or PTHrP 1-87, of inorganic pyrophosphate (PPi) elaboration suggested selective functional properties of the 1-173 isoform. Exposure to a neutralizing antibody to PTHrP increased PPi elaboration by articular chondrocytes. CONCLUSION: Increased expression of PTHrP, including the 1-173 isoform, has the potential to contribute to the pathologic differentiated functions of chondrocytes, including mineralization, in OA. PMID- 9870873 TI - Enhanced and coordinated in vivo expression of inflammatory cytokines and nitric oxide synthase by chondrocytes from patients with osteoarthritis. AB - OBJECTIVE: To evaluate the sites of expression of interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNFalpha), and inducible nitric oxide synthase (iNOS) in patients with inflammatory and degenerative joint diseases. METHODS: Cytokines and iNOS were detected by immunohistochemistry analysis of synovial and cartilage biopsy specimens obtained at knee arthroscopy in patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), osteoarthritis (OA), and traumatic knee arthritis. Cytokine and iNOS expression was quantified using computerized image analysis. RESULTS: IL-1beta, TNFalpha, and iNOS were highly expressed by synovial cells (lining layer cells, infiltrating leukocytes, endothelial cells) from patients with inflammatory arthritides and significantly less by synovial cells from patients with OA and traumatic arthritis. In contrast, the latter patients showed high chondrocyte expression of cytokines and iNOS while RA and PsA patients had only minor chondrocyte positivity. In both joint compartments, IL-1beta expression, TNFalpha expression, and iNOS expression were strongly correlated. CONCLUSION: The enhanced and coordinated expression of IL-1beta, TNFalpha, and iNOS by chondrocytes strongly supports the hypothesis that chondrocytes are the major site of production of mediators of inflammation in human OA, thus playing a primary role in the pathogenesis of this disease. PMID- 9870874 TI - Murine cytomegalovirus induces a Sjogren's syndrome-like disease in C57Bl/6 lpr/lpr mice. AB - OBJECTIVE: To analyze Fas and tumor necrosis factor receptor I (TNFRI) apoptosis pathways in salivary gland inflammatory disease induced by murine cytomegalovirus (MCMV) infection. METHODS: Four different strains of mice (C57BI/6 [B6]-+/+, Fas deficient B6-lpr/lpr, TNFRI-deficient B6-tnfr1(0/0), and B6-tnfr1(0/0)-lpr/lpr mice) were infected intraperitoneally with the Smith strain of MCMV (1 x 10(5) plaque-forming units). Viral load was determined by a plaque assay, inflammation and apoptosis by immunohistochemistry and staining with terminal dUTP nickend labeling, and autoantibodies by enzyme-linked immunosorbent assay. RESULTS: Infectious MCMV was not detectable by day 100. Although all MCMV-infected mice developed acute sialadenitis by day 28, a chronic (>100 days), severe salivary gland inflammation and anti-Ro and anti-La antibodies developed only in the B6 lpr/lpr mice. Apoptotic cells were detected during the acute, but not the chronic, phase of inflammation. CONCLUSION: Both Fas- and TNFRI-mediated apoptosis contribute to the clearance of MCMV-infected cells in the salivary glands. However, because Fas-mediated apoptosis is necessary for the down modulation of the immune response, a defect in this process can lead to a postinfection, chronic inflammatory response that resembles Sjogren's syndrome. PMID- 9870875 TI - Gelatin/chondroitin 6-sulfate microspheres for the delivery of therapeutic proteins to the joint. AB - OBJECTIVE: To develop a biodegradable, inflammation-responsive microsphere system for the intraarticular delivery of therapeutic proteins. METHODS: Microspheres were synthesized by complex coacervation. Radiolabeled protein release and microsphere degradation were assessed by exposing the microspheres to human synovial fluids (SF) and recombinant gelatinase. Microsphere degradation was confirmed by scanning electron microscopy (SEM). Microsphere biocompatibility was evaluated in vitro by incubating the microspheres with human synoviocytes, and in vivo by injection into mouse joints. RESULTS: Optimal microsphere formulation was developed. Significant (up to 100%) release of encapsulated protein occurred in SF samples with measurable metalloprotease activity, while release was minimal in SF with negligible activity. Dissolution of microspheres exposed to gelatinase was confirmed by SEM. Microspheres were found to be noncytotoxic in vitro, and noninflammatory in vivo. CONCLUSION: Microsphere encapsulation is an inflammation responsive and biocompatible system of protein delivery that holds promise for use in the delivery of therapeutic proteins to the joint. PMID- 9870876 TI - Treatment of rheumatoid arthritis with recombinant human interleukin-1 receptor antagonist. AB - OBJECTIVE: To evaluate the efficacy and safety of interleukin-1 receptor antagonist (IL-1Ra) in patients with rheumatoid arthritis (RA). METHODS: Patients with active and severe RA (disease duration <8 years) were recruited into a 24 week, double-blind, randomized, placebo-controlled, multicenter study. Doses of nonsteroidal antiinflammatory drugs and/or oral corticosteroids (< or =10 mg prednisolone daily) remained constant throughout the study. Any disease-modifying antirheumatic drugs that were being administered were discontinued at least 6 weeks prior to enrollment. Patients were randomized to 1 of 4 treatment groups: placebo or a single, self-administered subcutaneous injection of IL-1Ra at a daily dose of 30 mg, 75 mg, or 150 mg. RESULTS: A total of 472 patients were recruited. At enrollment, the mean age, sex ratio, disease duration, and percentage of patients with rheumatoid factor and erosions were similar in the 4 treatment groups. The clinical parameters of disease activity were similar in each treatment group and were consistent with active and severe RA. At 24 weeks, of the patients who received 150 mg/day IL-1Ra, 43% met the American College of Rheumatology criteria for response (the primary efficacy measure), 44% met the Paulus criteria, and statistically significant improvements were seen in the number of swollen joints, number of tender joints, investigator's assessment of disease activity, patient's assessment of disease activity, pain score on a visual analog scale, duration of morning stiffness, Health Assessment Questionnaire score, C-reactive protein level, and erythrocyte sedimentation rate. In addition, the rate of radiologic progression in the patients receiving IL-1Ra was significantly less than in the placebo group at 24 weeks, as evidenced by the Larsen score and the erosive joint count. IL-1Ra was well tolerated and no serious adverse events were observed. An injection-site reaction was the most frequently observed adverse event, and this resulted in a 5% rate of withdrawal from the study among those receiving IL-1Ra at 150 mg/day. CONCLUSION: This study confirmed both the efficacy and the safety of IL-1Ra in a large cohort of patients with active and severe RA. IL-1Ra is the first biologic agent to demonstrate a beneficial effect on the rate of joint erosion. PMID- 9870877 TI - Reduction of NOS2 overexpression in rheumatoid arthritis patients treated with anti-tumor necrosis factor alpha monoclonal antibody (cA2). AB - OBJECTIVE: Peripheral blood mononuclear cells (PBMC) from patients with rheumatoid arthritis (RA) have increased expression of nitric oxide synthase type 2 (NOS2) protein and enhanced formation of nitric oxide (NO) that correlate with disease activity. NO may play a role in the inflammation of RA. Treatment of RA patients with a chimeric monoclonal antibody against tumor necrosis factor alpha (TNFalpha; cA2) results in clinical improvement in the majority of patients. The present study was designed to determine if cA2 therapy decreases PBMC NOS2 protein expression and NOS enzyme activity in RA patients. METHODS: RA patients receiving background oral methotrexate participated in a double-blind, placebo controlled clinical trial in which they were randomly assigned to receive a single infusion of either placebo or cA2 at 5, 10, or 20 mg/kg. NOS2 protein and NOS enzyme activity were measured in PBMC at baseline and 4 weeks following cA2 therapy. These results were compared with the degree of clinical change in disease activity. RESULTS: At baseline, elevated levels of NOS2 protein and NOS enzyme activity were more frequently detected in PBMC from RA patients than in those from healthy controls. Treatment of the RA patients with cA2 significantly reduced NOS2 protein expression and NOS enzyme activity. Changes in NOS activity following treatment correlated significantly with changes in the number of tender joints. CONCLUSION: These results indicate that TNFalpha likely plays an important role in enhancing NOS2 expression in RA, and that the antiinflammatory effects of cA2 treatment may be mediated by a reduction of NO overproduction. PMID- 9870878 TI - Functional analysis of rheumatoid factor-producing B cells from the synovial fluid of rheumatoid arthritis patients. AB - OBJECTIVE: To understand the regulation of rheumatoid factor (RF) production in rheumatoid arthritis (RA), we studied IgM-RF production by B cells isolated from the synovial fluid (SF). METHODS: Highly purified SF and peripheral blood (PB) B cells were isolated by negative selection in a fluorescence-activated cell sorter (FACS) and then cultured with either L cells, CD40 ligand (CD40L)-transfected L cells, or type B synoviocytes in the presence or absence of interleukin-2 (IL-2), IL-4, or IL-10. Total IgM and IgM-RF were detected after 14 days by enzyme-linked immunosorbent assay. Enzyme-linked immunospot assays were performed to detect cells that spontaneously produced immunoglobulin. SF B cells were also phenotypically characterized by FACS analysis. RESULTS: Terminally differentiated CD20-,CD38+ synovial plasma cells (PC) present in the SF of RA patients secreted IgM-RF in the absence of a stimulus. IgM-RF production markedly increased when SF B cells were cultured in the presence of type B RA synoviocytes together with IL 10, but independently of CD40-CD40L interaction. Although CD20-,CD38+ PC could also be demonstrated in SF B cells from patients with other forms of arthritis, IgM-RF production was restricted to the SF B cell cultures of patients with seropositive RA. The frequency of IgM-RF-producing cells among IgM-producing PC in patients with seropositive RA was estimated to be as much as 50%. CONCLUSION: These data demonstrate that terminally differentiated CD20-,CD38+ IgM-RF producing B cells are specifically present in the inflamed joints of patients with seropositive RA. There is evidence that the local environment in the rheumatoid joint favors RF production. The relatively high frequency of IgM-RF PC in the SF B cell population provides evidence of a dominant RA-specific antigen driven response in the development of the synovial PC repertoire. PMID- 9870879 TI - Cytokine-regulated expression of activated leukocyte cell adhesion molecule (CD166) on monocyte-lineage cells and in rheumatoid arthritis synovium. AB - OBJECTIVE: To determine whether monocyte/macrophage expression of the CD6 ligand, activated leukocyte cell adhesion molecule (ALCAM) (CD166), is regulated by cytokines during inflammation in rheumatoid arthritis (RA). METHODS: We used flow cytometry to test whether cytokines present in rheumatoid synovium could regulate ALCAM cell surface expression on peripheral blood (PB) monocytes and RA synovial fluid (SF) macrophages, and we examined ALCAM expression in situ in RA synovium by immunofluorescence. RESULTS: The monocyte differentiation factors interleukin 3, macrophage colony-stimulating factor (M-CSF), and granulocyte-macrophage colony-stimulating factor augmented ALCAM expression on PB monocytes. ALCAM was expressed on monocyte-lineage cells in situ in inflamed synovium from patients with RA (9 of 9), but not in uninflamed synovium from patients with joint trauma (0 of 3). Furthermore, in vitro culture-induced ALCAM expression on PB monocytes and CD14+ RA SF cells was inhibited by an M-CSF neutralizing antibody. CONCLUSION: ALCAM expression on PB and SF monocytes/macrophages is enhanced by M CSF. PMID- 9870880 TI - Predictors of subjective fatigue among individuals with rheumatoid arthritis. AB - OBJECTIVE: To examine a range of variables potentially associated with rheumatoid arthritis (RA)-related fatigue and to determine which variables best predict subjective fatigue. METHODS: Measures of fatigue, disease activity, pain, and various psychosocial factors were gathered from 73 individuals with RA. Correlations between fatigue and other variables were examined, and the "best" predictors of fatigue were determined with multiple regression analyses. RESULTS: Many of the variables that were significantly correlated with fatigue had a psychosocial character. Moreover, the "best" predictors of increased fatigue were higher levels of pain, more depressive symptoms, and female sex. Longer symptom duration, less perceived adequacy of social support, and less disease activity were significant predictors of fatigue, over and above the "best" model. CONCLUSION: RA-related fatigue appears to be strongly associated with psychosocial variables, apart from disease activity per se. Correspondingly, treatment of fatigue may be enhanced by interventions that address relevant cognitive and behavioral dimensions. PMID- 9870881 TI - Detection of anti-Ro/SSA and anti-La/SSB autoantibody-producing cells in salivary glands from patients with Sjogren's syndrome. AB - OBJECTIVE: To investigate and identify the presence of cells producing anti Ro/SSA and anti-La/SSB autoantibodies in salivary glands from patients with Sjogren's syndrome (SS). METHODS: Submucosal salivary gland biopsy samples from 10 SS patients (8 with and 2 without circulating Ro and La autoantibodies) and 14 control subjects were evaluated. Frozen tissue sections were immunostained by an avidin-biotin complex technique, using biotinylated recombinant Ro and La proteins as detection reagents. Autoantibody levels in SS patient sera were analyzed by enzyme-linked immunosorbent assay. RESULTS: Cells producing autoantibodies to the Ro 52-kd, Ro 60-kd, and La proteins were recorded in 8, 6, and 7 of the 10 SS patient biopsy samples, respectively. Samples from the 2 SS patients without circulating Ro and La autoantibodies were negative for these autoantibody-producing cells, as were all control biopsy samples. A strong positive correlation between the presence of autoantibodies in sera and the presence of autoantibody-producing cells in glandular biopsy tissues was evident. The number of autoantibody-producing cells and the serum autoantibody levels were also correlated (r(s)=0.94, P < 0.0001). CONCLUSION: Using a novel technique, we have demonstrated the presence of Ro and La autoantibody-producing cells in salivary gland biopsy tissues from patients with SS. These findings indicate that anti-Ro/ SSA and anti-La/SSB autoantibodies are produced and are present at sites of inflammation and indicate their potential involvement in the autoimmune exocrinopathy of this disease. PMID- 9870882 TI - The impact of hemiparalysis on the expression of osteoarthritis. AB - OBJECTIVE: Primary generalized osteoarthritis (OA), the most prevalent joint disease, is usually symmetric. Sporadic case reports mention decreased OA manifestations in limbs in which there are neurologic deficits, but no systematic research has been published. The aim of the present study was to examine these observations in a planned and controlled survey in a group of patients with OA. METHODS: Seventy-five geriatric patients with a history of stroke and hemiparalysis were studied clinically and radiographically (hand radiographs; graded according to a modified Altman method) for the presence and the degree of OA in the hands. Detailed clinical and radiologic scores were calculated for each hand. Demographic, occupational, and neurologic data were collected. Patients with other joint or neurologic conditions were excluded. A group of 55 elderly patients without stroke were similarly studied (controls). Scores in the paralyzed hand were compared with those in the nonparalyzed hand in the stroke patients and subgroups (by Student's paired t-test and Wilcoxon test). Scores in the dominant hands were compared with those in the nondominant hands in stroke patients and control subjects (by Student's paired t-test and Mann-Whitney test). Correlation between the degree of neurologic damage and OA asymmetry (Pearson's correlation coefficient) was also sought. RESULTS: Paralyzed hands showed significantly fewer OA changes than nonparalyzed hands, both clinically and radiologically. This trend, accentuated in patients with more severe paralysis, disappeared in those with mild residual paresis. Asymmetry of OA was more pronounced in patients with flaccid, compared with spastic, paralysis. The degree of paralysis and loss of muscle strength correlated with the degree of OA asymmetry. Women had significantly higher OA scores than men. In the control group, dominant hands had higher OA scores, but this finding was concealed among hemiparalyzed patients. Lifetime gross occupational load and present grip strength did not correlate with the degree of OA. CONCLUSION: In elderly patients, hemiparalysis reduces ipsilateral hand expression of OA, while OA is accentuated (or increased) in the dominant hand of patients without paralysis. This first systematic study confirms the findings of previous case reports and lends support to the role of biomechanical factors in the development of OA. PMID- 9870883 TI - Wegener's granulomatosis: patient-reported effects of disease on health, function, and income. AB - OBJECTIVE: To evaluate the patient-perceived effects of Wegener's granulomatosis (WG) on health, function, income, and interpersonal relationships. METHODS: A self-administered questionnaire, originally designed by the authors and subsequently revised with the aid of a patient focus group, was completed by 60 patients with well-defined features of WG. Patients had WG for a median period of 5 years. RESULTS: Patients with chronic WG experienced substantial medical and functional morbidity and incurred significant socioeconomic losses. A prolonged delay in diagnosis (mean 16.8 months) and the need for multiple consultations prior to initiation of therapy may have contributed to medical morbidity. Although 73% of patients perceived their disease to be in remission following therapy, 78% of these patients required continuing immunosuppressive treatment many years after diagnosis. Eighty percent of patients reported that their normal activities of daily living were compromised. Half of those who were employed prior to diagnosis were required to modify their job or accept total disability (31%). A 26% (median) reduction in income within 1 year after diagnosis was reported. The effects of the disease on interpersonal relationships with a patient's spouse, family, and friends varied considerably. CONCLUSION: Advances in medical care have, for most patients, transformed WG from being a disease with a high potential for short-term mortality to being a chronic illness. This is the first study that has evaluated patients' assessments of the medical, socioeconomic, and quality of life effects of WG and its treatment. The effects of mortality, disability, and outpatient medical expenses indicate that the financial impact alone substantially exceeds prior estimates of $30 million per year in charges for hospitalizations in the US. PMID- 9870884 TI - The Bath Ankylosing Spondylitis Radiology Index (BASRI): a new, validated approach to disease assessment. AB - OBJECTIVE: To develop a reproducible and simple radiologic scoring system for the spine in patients with ankylosing spondylitis (AS): the Bath Ankylosing Spondylitis Radiology Index for the spine (BASRI-s). METHODS: Radiographs of 470 patients with AS were scored using the New York criteria for the sacroiliac joints and, similarly, grading the lumbar and cervical spine on a scale of 0-4 (for normal, suspicious, mild, moderate, and severe). These 3 scores were added together to produce the BASRI-s score (scored 2-12). Radiographs of 188 patients were used to test reproducibility. Blinded radiographs of 89 non-AS patients were included, randomly, to assess disease specificity. Sensitivity to change was assessed using 177 radiographs from 58 AS patients. RESULTS: Intra- and interobserver variation showed 75-86% and 73-79% complete agreement at all sites, respectively. Specificities of 0.83-0.89 suggested that the lumbar and cervical spine BASRI scores were disease specific. Sensitivity to change became apparent at 2 years (P < 0.001). Using a lateral view and an anteroposterior view of the lumbar spine was more sensitive than using a lateral view alone. Grading a set of radiographs (sacroiliac joints, lumbar spine, and cervical spine) took 30 seconds. CONCLUSION: BASRI is a reliable method for grading radiographic changes in patients with AS. It is disease specific, sensitive to change, valid, simple, and rapid to perform. PMID- 9870885 TI - Malignancy-associated remission of systemic lupus erythematosus maintained by autologous peripheral blood stem cell transplantation. AB - Many investigators worldwide are currently exploring the role of peripheral blood stem cell transplantation (PBSCT) in managing autoimmune diseases. We report the case of a woman with systemic lupus erythematosus (SLE) with mucocutaneous and renal involvement, who underwent PBSCT for stage IVB Hodgkin's disease. Following the development of the lymphoma, she has had a prolonged clinical and serologic remission of the SLE. The potential effects of lymphoproliferative disorders and PBSCT on the course of SLE are considered. PMID- 9870886 TI - Thalidomide induces amenorrhea in patients with lupus disease. PMID- 9870887 TI - Osteolytic lesions in T cell lymphoma. PMID- 9870888 TI - Assessing lupus. PMID- 9870889 TI - Broad-range bacterial polymerase chain reaction for identification of bacteria in inflamed joints: comment on the article by Wilbrink et al. PMID- 9870890 TI - "Disablement" framework variables in the assessment of juvenile arthritis outcomes: comment on the article by Giannini et al. PMID- 9870891 TI - New DNA diagnostic system for detection of factor V Leiden. AB - Use was made of allele-specific PCR to develop a highly effective DNA diagnostic system for detection of the factor V Leiden mutation in exon 10 of human factor V gene. The allele-specific primers contain a 3'-OH end nucleotide, which matches a mutant or wild-type nucleotide of the template DNA (A-allele and G-allele, respectively) and also one mismatched nucleotide near the 3'-end. The universal primers have an internal mismatch with the mutant nucleotide of the template DNA and another mismatched nucleotide at 3'-OH end. The A-allele-specific primer enables the preferential amplification of both the homozygous and heterozygous mutant alleles. The extension of the G-allele-specific primer or the universal one is inhibited in the presence of the homozygous factor V Leiden. The developed assay system allowed us to detect five patients, who are heterozygous for factor V Leiden among the 48 patients with deep venous thrombosis and pulmonary thromboembolism. PMID- 9870892 TI - Synergistic antithrombotic effects of argatroban and ticlopidine in the rat venous thrombosis model. AB - Argatroban, a synthetic thrombin inhibitor, and ticlopidine, an anti-platelet agent, are major antithrombotic agents. We investigated the antithrombotic effects of a combination of argatroban and ticlopidine in the rat venous thrombosis model. Argatroban or ticlopidine inhibited thrombus formation in a dose-dependent manner; 50% inhibition (ED50) is obtained with 1.0 mg/kg/h (infusion) argatroban or 30 mg/kg (p.o.) ticlopidine. The combination of argatroban and ticlopidine inhibited thrombus formation in a dose-dependent manner; ED50 is obtained with 0.25 mg/kg/h argatroban plus 10 mg/kg ticlopidine and 0.5 mg/kg/h argatroban plus 3 mg/kg ticlopidine, whereas 0.5 mg/kg/h argatroban alone or 10 mg/kg ticlopidine alone had negligible effect (<20% inhibition). Isobole analysis showed that the antithrombotic effects of the combination of argatroban and ticlopidine involved synergism with potentiation. In contrast, the combination of argatroban and ticlopidine did not prolong the bleeding time synergistically. These data showed that the combination therapy of argatroban and ticlopidine should be clinically beneficial, but the different administration route may restrict the clinical usage. PMID- 9870893 TI - Activation of the haemostatic system in children with juvenile rheumatoid arthritis correlates with disease activity. AB - Twenty-four children with juvenile rheumatoid arthritis (JRA) and 10 children with postinfectious arthropathies were investigated for markers of blood coagulation and fibrinolytic activity: Prothrombin fragment 1+2 (F1+2), thrombin antithrombin complex (TAT), and D-Dimer were measured using solid phase enzyme linked immunosorbent assays (ELISA). Results were compared with clinical and conventional laboratory signs of disease activity. F1+2, TAT, D-Dimer, and fibrinogen were significantly elevated in children with JRA as compared with healthy children and children with postinfectious arthropathies. F1+2, TAT, and D Dimer correlated significantly with disease activity, assessed by determination of the joint index score and C-reactive protein (CRP). The study demonstrates a subclinical activation of the haemostatic system in children with JRA correlating with disease activity, which might be caused by the action of several immunomediators on cells (monocytes, endothelial cells) playing a role in the regulation of blood coagulation activity. PMID- 9870895 TI - Oxygen free radical-dependent increased platelet function in beta-thalassemia major patients. PMID- 9870894 TI - Chemical sulfonation and anticoagulant activity of acharan sulfate. AB - Acharan sulfate is a glycosaminoglycan prepared from the giant African snail, Achatina fulica. This polysaccharide has a repeating disaccharide structure of - >4)-2-deoxy-2-acetamido-alpha-D-glucopyranose (1-->4)-2-sulfo-alpha-L idopyranosyluronic acid (1-->). Its structure is related to heparin and heparan sulfate but is distinctly different from all known members of these classes of glycosaminoglycans. Because of its structural similarities to heparin, chemically modified acharan sulfate was studied to understand the chemical structure effected its anticoagulant activity. After de-N-acetylation, acharan sulfate was N-sulfonated using either chlorosulfonic acid-pyridine or sulfur trioxide trimethylamine complex. The sulfate level in these products ranged from 22 to 24%(w/w), significantly less than that of heparin at 36%. The molecular weight of both N-sulfoacharan sulfates were comparable with that of heparin. In vitro anticoagulant activity assays showed that N-sulfoacharan sulfate derivatives were moderately active for the inhibition of thrombin and neither product showed any measurable anti-factor Xa activity. The differences in the activities of N sulfoacharan sulfates produced by these two methods are probably ascribable to a small level of concomitant O-sulfonation obtained when using chlorosulfonic acid pyridine. PMID- 9870896 TI - Precision of patients' measurements of the international normalized ratio (INR) using a patient operated whole blood home coagulometer. PMID- 9870897 TI - The pharmacokinetics of the kininogens. AB - Recent evidence has shown that plasma high molecular weight kininogen and both kininogens have the ability to modulate prekallikrein activation and thrombin induced platelet activation, respectively. However, nothing is known about the plasma clearance and tissue distribution of these proteins. We examined the in vivo pharmacokinetics of high (HK) and low (LK) molecular weight kininogens in rats. 125I-HK and -LK molecular weight kininogens' clearance in rats best-fitted a biexponential model. For HK, the t1/2alpha and t1/2beta were 0.6 and 9.5 h and for LK, 0.78 and 7.4 h, respectively. 125I-kinin-free HK (cleaved HK) was cleared with a t1/2alpha and t1/2beta of 0.45 and 9.9 h, respectively. 125I-Domain 3 of kininogens was cleared with a t1/2beta and t1/2c of 0.99 and 13.3 h, respectively. HK was mostly concentrated in lung; LK, domain 3, and cleaved HK were mostly concentrated in kidney. The kininogens were also concentrated in liver, spleen, and skin. These studies indicate that protein size rather than form is the major determinant of its clearance. Furthermore, the distribution of the kininogens is where bradykinin metabolism and activity are well described. PMID- 9870898 TI - Activated protein C (APC) resistance in indian juvenile deep vein thrombosis (DVT) PMID- 9870901 TI - Acid-catalyzed isomerization of fucosterol and delta5-avenasterol. AB - This work shows that fucosterol, delta5-avenasterol, and similar ethylidene-side chain sterols can undergo acid-catalyzed isomerization to give a mixture of five isomers. Four isomers formed from fucosterol were analyzed, using gas chromatography-mass spectrometry, and were characterized as delta5-avenasterol, two delta5,23-stigmastadienols, and delta5,24(25)-stigmastadienol. When the unsaponifiables fraction from oat oil was subjected to acid hydrolysis, the two delta5,23-stigmastadienol isomers and delta5,24(25)-stigmastadienol were detected while fucosterol coeluted with sitosterol. Interisomerization of ethylidene-side chain sterols represents a limitation to the use of the acid hydrolysis method in the determination of sterols in food and other plant materials rich in these sterols, e.g., oat lipids. PMID- 9870900 TI - A pathway for biosynthesis of divinyl ether fatty acids in green leaves. AB - [1-14C]alpha-Linolenic acid was incubated with a particulate fraction of homogenate of leaves of the meadow buttercup (Ranunculus acris L.). The main product was a divinyl ether fatty acid, which was identified as 12-[1'(Z),3'(Z) hexadienyloxy]-9(Z),11(E)-dodecadienoic acid. Addition of glutathione peroxidase and reduced glutathione to incubations of alpha-linolenic acid almost completely suppressed formation of the divinyl ether acid and resulted in the appearance of 13(S)-hydroxy-9(Z), 11(E),15(Z)-octadecatrienoic acid as the main product. This result, together with the finding that 13(S)-hydroperoxy-9(Z), 11(E),15(Z) octadecatrienoic acid served as an efficient precursor of the divinyl ether fatty acid, indicated that divinyl ether biosynthesis in leaves of R. acris occurred by a two-step pathway involving an omega6-lipoxygenase and a divinyl ether synthase. Incubations of isomeric hydroperoxides derived from alpha-linolenic and linoleic acids with the enzyme preparation from R. acris showed that 13(S)-hydroperoxy 9(Z),11(E)-octadecadienoic acid was transformed into the divinyl ether 12-[1'(Z) hexenyloxy]-9(Z), 11(E)-dodecadienoic acid. In contrast, neither the 9(S) hydroperoxides of linoleic or alpha-linolenic acids nor the 13(R)-hydroperoxide of alpha-linolenic acid served as precursors of divinyl ethers. PMID- 9870899 TI - Obesity as a risk factor for certain types of cancer. AB - In conclusion, obesity has been associated with increased risk for a number of different types of cancer. The evidence has been most consistent for endometrial cancer, breast cancer in postmenopausal women, and renal cell cancer. More variable results have been reported for colorectal, prostate and pancreatic cancer. Possible mechanisms by which obesity may influence cancer risk include alteration in hormonal patterns, including sex hormones and insulin, and factors such as the distribution of body fat and changes in adiposity at different ages. The increasing prevalence of obesity in many parts of the world emphasizes the importance of learning more about the relationship between obesity and cancer and the mechanisms involved in their interaction. PMID- 9870902 TI - Surface pressure-dependent cross-modulation of sphingomyelinase and phospholipase A2 in monolayers. AB - We investigated the ways in which phospholipase A2 and sphingomyelinase are mutually modulated at lipid interfaces. The activity of one enzyme is affected by its own reaction products and by substrates and products of the other enzyme; all this depends differently on the lateral surface pressure. Ceramide inhibits both the sphingomyelinase activity rate and the extent of degradation, and decreases the lag time at all surface pressures. Dilauroyl- and dipalmitoylphosphatidylcholine, the substrates of phospholipase A2 (PLA2), do not affect sphingomyelinase activity. The products of PLA2, palmitic acid and lysopalmitoylphosphatidylcholine, strongly enhance and shift to high surface pressures the activity optimum and the cutoff point of sphingomyelinase. Palmitic acid also shifts to high surface pressures the cut-off point of PLA2 activity. Sphingomyelin strongly inhibits PLA2 at surface pressures above 5 mN/m, while ceramide shifts the cut-off point and the activity optimum to high surface pressures. The sphingolipids increase the lag time of PLA2 at low surface pressures. Both phosphohydrolytic pathways involve different levels of control on precatalytic steps and on the rate of activity that appear independent on specific alterations of molecular packing and surface potential. The mutual lipid mediated interfacial modulation between both phosphohydrolytic pathways indicates that phospholipid degradation may be self-amplified or dampened depending on subtle changes of surface pressure and composition. PMID- 9870903 TI - GD3 and GM2 synthase activities in rat testes during the period of sexual development. AB - Activities of two key enzymes of gangliosides biosynthesis were determined in rat testes during development. GD3 synthase activity was low and showed small variations with age. GM2 synthase activity increased 10-fold in testes from 10- to 30-d-old animals, showing a maximum activity at 30 d, followed by a small decrease until 45 d and then a constant activity up to adulthood. These developmental changes in the activity of both glycosyltransferases were related to the increasing complexity in the ganglioside pattern observed in rats testes during the period of sexual development. PMID- 9870904 TI - Lipid and fatty acid composition of brush border membrane of rat intestine during starvation. AB - Alterations in the lipid and fatty acid composition of brush border membrane (BBM) of small intestine were studied in well-fed, starved, and refed rats. The ratios of cholesterol/phospholipid (mol/mol), sphingomyelin/phosphatidylcholine (mol/mol), protein/lipid (w/w), and free fatty acids (w/w) decreased whereas the total phospholipid (w/w) ratio and the double-bond index increased in BBM of the intestine of the starved rat compared to that of the well-fed rat. Analyses of fatty acids showed higher percentage of stearic and arachidonic acids whereas oleic and linoleic acids decreased under starvation. The acyl chain of starved rat BBM was less ordered compared with that of well-fed rat BBM. On refeeding, these changes were restored to well-fed levels. The change in membrane state under starvation is associated with alterations in the lipid and fatty acid composition of BBM and may be responsible for functional changes that occur under nutritional stress. PMID- 9870905 TI - Metabolism of dietary alpha-linolenic acid vs. eicosapentaenoic acid in rat immune cell phospholipids during endotoxemia. AB - Short-term (i.e., 3 d) continuous enteral feeding of diets containing eicosapentaenoic (EPA) and gamma-linolenic (GLA) polyunsaturated fatty acids (PUFA) to endotoxemic rats reduces the levels of arachidonic acid (AA) and linoleic acid (LA) in alveolar macrophage (AM) and liver Kupffer and endothelial (K&E) cell phospholipids with attendant decreases in prostaglandin formation by these cells in vitro. Diets that contain alpha-linolenic acid (LNA) as a substrate for endogenous formation of EPA may not be as effective in facilitating these immune cell modifications given the limited activity of delta6 desaturase. In the present study we compared the effectiveness of an LNA-enriched diet vs. an (EPA + GLA)-enriched diet to displace phospholipid AA from AM and liver K&E cells in vivo in endotoxemic rats fed enterally for 3 or 6 d. We determined the fatty acid composition of AM and K&E cell phospholipids by gas chromatography. We found that AM and K&E cells from rats that had received the EPA + GLA diet for 3 d had significantly (P < 0.001) higher mole percentage of EPA and the GLA metabolite, dihomoGLA, than corresponding cells from rats given the LNA diet or a control diet enriched with LA. Rats given the LNA diet had relatively low levels of stearidonic acid, EPA and other n-3 PUFA, while rats given the LA diet had low levels of GLA and dihomoGLA. We conclude that diets enriched with LNA or LA may not be as effective as those enriched with EPA + GLA for purposes of fostering incorporation of EPA or dihomoGLA into and displacement of AA from macrophage phospholipids under pathophysiologic conditions commonly found in acutely septic patients. PMID- 9870907 TI - Low density lipoprotein of synovial fluid in inflammatory joint disease is mildly oxidized. AB - Oxidatively modified low density lipoprotein (LDL) has many biological activities which could contribute to the pathology of the atherosclerotic lesion. Because atherosclerosis has an inflammatory component, there has been much interest in the extent to which LDL could be oxidatively modified in vivo by inflammation. The present study examined LDL present in an accessible inflammatory site, the inflamed synovial joint, for evidence of compositional change and oxidative modification. LDL was isolated from knee joint synovial fluid (SF) from subjects with inflammatory arthropathies and also from matched plasma samples. SF and plasma LDL had similar free cholesterol and alpha-tocopherol content, but SF LDL had a lower content of esterified cholesterol. On electrophoresis, SF LDL was slightly more electronegative than LDL from matched plasma samples, but the changes were much less than those resulting from Cu2+-treatment of LDL. Oxidized cholesterol was not detected in any samples, but cholesterol ester hydroperoxide levels were greater in SF than in plasma LDL. When samples from three subjects were incubated with macrophages, the SF LDL did not cause significant loading of the cells with cholesterol or cholesterol esters, in contrast to the situation with acetylated LDL. Overall, the SF LDL displayed evidence of slightly increased oxidation by comparison with matched plasma samples. Despite their isolation from an environment with active inflammation, changes were modest compared with those resulting from Cu2+ treatment. Thus, extensive LDL oxidation is not a necessary correlate of location in a chronic inflammatory site, even though it is characteristic of atherosclerotic lesions. PMID- 9870906 TI - Effect of tetracosahexaenoic acid on the content and release of histamine, and eicosanoid production in MC/9 mouse mast cell. AB - 6,9,12,15,18,21-Tetracosahexaenoic acid (24:6n-3) was isolated from a brittle star, Ophiura sarsi Lutken, at >95% purity to evaluate its physiological functions. The effects of 24:6n-3 on the production of leukotriene (LT)-related compounds such as LTB4, LTC4 and 5-hydroxyeicosatetraenoic acid, and the accumulation and release of histamine in an MC/9 mouse mast cell line were studied. We found that 24:6n-3 could inhibit the antigen-stimulated production of LT-related compounds as well as other n-3 polyunsaturated fatty acids (PUFA) such as eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3), which are major n-3 PUFA in fish oils; 24:6n-3 was also shown to reduce the histamine content in MC/9 cells at 25 microM (27% reduction from the control), and the effect was diminished with increase of the fatty acid concentration (up to 100 microM). These two n-3 PUFA, 20:5n-3 and 22:6n-3, also reduced the histamine content (16 and 20% reduction at 25 microM, respectively), whereas arachidonic acid (20:4n-6) increased it (18% increase at 25 microM). Spontaneous- and antigen induced release of histamine was not influenced with these PUFA (at 25 microM). Ionophore-stimulated release of histamine was suppressed by the PUFA (13, 9, 15, and 11% reduction with 20:4n-6, 20:5n-3, 22:6n-3, and 24:6n-3, respectively). The patterns of the effects of 24:6n-3 on the synthesis of eicosanoids and histamine content were more similar to those of 22:6n-3 than 20:5n-3. From these results, 24:6n-3 can be expected to have anti-inflammatory activity and antiallergic activities similar to those of 22:6n-3. PMID- 9870909 TI - Low doses of eicosapentaenoic acid, docosahexaenoic acid, and hypolipidemic eicosapentaenoic acid derivatives have no effect on lipid peroxidation in plasma. AB - It was of interest to investigate the influence of both high doses of eicosapentaenoic acid (EPA) and low doses of 2- or 3-methylated EPA on the antioxidant status, as they all cause hypolipidemia, but the dose required is quite different. We fed low doses (250 mg/d/kg body wt) of different EPA derivatives or high doses (1500 mg/d/kg body wt) of EPA and DHA to rats for 5 and 7 d, respectively. The most potent hypolipidemic EPA derivative, 2,2-dimethyl EPA, did not change the malondialdehyde content in liver or plasma. Plasma vitamin E decreased only after supplementation of those EPA derivatives that caused the greatest increase in the fatty acyl-CoA oxidase activity. Fatty acyl CoA oxidase activity increased after administration of both EPA and DHA at high doses. High doses of EPA and DHA decreased plasma vitamin E content, whereas only DHA elevated lipid peroxidation. In liver, however, both EPA and DHA increased lipid peroxidation, but the hepatic level of vitamin E was unchanged. The glutathione-requiring enzymes and the glutathione level were unaffected, and no significant changes in the activities of xanthine oxidase and superoxide dismutase were observed in either low- or high-dose experiments. In conclusion, increased peroxisomal beta-oxidation in combination with high amounts of polyunsaturated fatty acids caused elevated lipid peroxidation. At low doses of polyunsaturated fatty acids, lipid peroxidation was unchanged, in spite of increased peroxisomal beta-oxidation, indicating that polyunsaturation is the most important factor for lipid peroxidation. PMID- 9870908 TI - Lipid peroxidation of isolated chylomicrons and oxidative status in plasma after intake of highly purified eicosapentaenoic or docosahexaenoic acids. AB - Fourteen healthy male volunteers were given two separate high-saturated-fat meals with and without the addition of 4 g highly purified ethyl esters of either eicosapentaenoic acid (EPA) (95% pure, n = 7) or docosahexaenoic acid (DHA) (90% pure, n = 7) supplied as 1-g capsules each containing 3.4 mg vitamin E. The chylomicrons were isolated 6 h after the meals, at peak concentrations of n-3 fatty acids (FA). Addition of n-3 FA with the meal caused a 10.4-fold increase in the concentration of n-3 FA in chylomicrons compared to the saturated fat meal without addition of n-3 FA. After the saturated-fat meal, the concentration of thiobarbituric acid-reactive substances (TBARS) was 327.6 +/- 34.6 nmol/mmol triacylglycerol (TAG), which increased to 1015.8 +/- 212.0 nmol/mmol TAG (P < 0.0001, n = 14) after EPA and DHA were added to the meal. There was no significant correlation between the concentrations of TBARS and vitamin E in the chylomicrons collected 6 h after the test meal. The present findings demonstrate an immediate increase in chylomicron peroxidation ex vivo provided by intake of highly purified n-3 FA. The capsular content of vitamin E was absorbed into chylomicrons, but the amount of vitamin E was apparently not sufficient to protect chylomicrons against lipid peroxidation ex vivo. Daily intake of 4 g n-3 FA either as EPA or DHA for 5 wk did not change the plasma concentration of TBARS. Although not significantly different between groups, DHA supplementation decreased total glutathione in plasma (P < 0.05) and EPA supplementation increased plasma concentration of vitamin E (P < 0.05). The other lipid-soluble and polar antioxidants in plasma remained unchanged during 5 wk of intervention with highly purified n-3 FA. PMID- 9870910 TI - Quantitative determination of butylated hydroxyanisole, butylated hydroxytoluene, and tert-butyl hydroquinone in oils, foods, and biological fluids by high performance liquid chromatography with fluorometric detection. AB - Concentrations of synthetic antioxidants butylated hydroxyanisole, butylated hydroxytoluene, and tert-butyl hydroquinone were quantified using a high performance liquid chromatograph with spectrofluorometric detector. The antioxidants were separated and eluted on a reversed-phase column by gradient of a mixture of H2O/acetonitrile/acetic acid (66.5: 28.5:5, by vol) and a mixture of acetonitrile/acetic acid (95:5, vol/vol). The eluants were monitored at emission and excitation wavelengths of 310 and 280 nm, respectively. Calibration curves obtained using peak areas against concentration showed high coefficients of multiple determination (R2 > 0.99) for all antioxidants. Known concentrations of added antioxidant standards were recoverable within 98-99% from oils and over 93% from mouse blood. This method requires minimum sample extraction and purification before analysis and provides a relatively high percentage recovery. The method has been applied successfully for the measurement of antioxidant concentrations in oils, dried foods, and biological fluids. PMID- 9870911 TI - CD23 and allergic pulmonary inflammation: potential role as an inhibitor. AB - CD23, a receptor for immunoglobulin E, is expressed at increased levels in asthmatic and atopic individuals and has been associated with disorders characterized by chronic inflammation. Using an established murine model, we employed several complementary strategies to investigate the role of CD23 in allergic pulmonary inflammation and airway hyperresponsiveness (AHR). Specifically, these approaches included the modulation of CD23 function in vivo by administration of anti-CD23 monoclonal antibody (mAb) or Fab fragments to wild type mice and the analysis of CD23-deficient mice. Administration of anti-CD23 mAb, but not anti-CD23 Fab fragments, produced attenuation of pulmonary inflammation, AHR, and CD8(+) T-cell activation. On the basis of a model that the anti-CD23 mAb transduces, whereas the Fab fragment inhibits, CD23 signaling, these results suggest that CD23 negatively regulates pulmonary inflammation and AHR. This hypothesis is supported by our observation that CD23-deficient mice developed increased inflammation and AHR after sensitization and challenge with allergen. Together, these results indicate that CD23 negatively regulates pulmonary inflammation and airway hyperreactivity. PMID- 9870912 TI - CD34(+)/interleukin-5Ralpha messenger RNA+ cells in the bronchial mucosa in asthma: potential airway eosinophil progenitors. AB - Eosinophil differentiation is thought to occur by the action of interleukin (IL) 5 on CD34(+) progenitor cells. The allergen-induced increase in eosinophil numbers in isolated airway preparations in vitro, and detection of increased numbers of circulating CD34(+) cells in atopic subjects, led us to the hypothesis that the eosinophil infiltration of the airway in asthma may result from local mucosal differentiation, in addition to recruitment from the bone marrow. We examined CD34(+) cell numbers by immunohistochemistry and IL-5 receptor alpha (IL 5Ralpha) messenger RNA (mRNA) expression by in situ hybridization in bronchial biopsies from atopic asthmatic patients, and from atopic and nonatopic control subjects. CD34(+) cell numbers were increased in the airway in atopic asthmatic and atopic nonasthmatic subjects. In contrast, CD34(+)/ IL-5Ralpha mRNA+ cells were increased in asthmatic subjects when compared with both atopic and nonatopic control subjects. Airway numbers of CD34(+)/IL-5Ralpha mRNA+ cells were correlated to airway caliber in asthmatic subjects and to eosinophil numbers. These findings support the concept that eosinophils may differentiate locally in the airway in asthma. PMID- 9870913 TI - Vascular endothelial growth factor gene expression in human fetal lung in vitro. AB - Neonatal respiratory function depends on the development of a well-formed pulmonary capillary bed. Vascular endothelial growth factor (VEGF) is a potent inducer of endothelial cell growth and angiogenesis. High levels of VEGF protein and messenger RNA (mRNA) have been detected in the developing lung, suggesting that VEGF plays a role in the development of the pulmonary capillary bed. To begin to understand the role of VEGF in human lung development, we explored the regulation of VEGF gene expression and the localization of VEGF protein and mRNA in a model of the developing human lung. VEGF protein and mRNA were detected in midtrimester human fetal lung tissue, and their levels increased with time in explant culture. VEGF protein and mRNA were increased by the maintenance of human fetal lung explants in 2% O2 environments compared with 20% O2 environments. VEGF mRNA levels were found to be increased by cyclic adenosine monophosphate (cAMP) in explants that were incubated in 20% O2, but not in those incubated in 2% O2. Immunostaining for VEGF protein demonstrated localization primarily in airway epithelial cells in midtrimester human fetal lung tissue. Immunostaining for VEGF increased with incubation of human fetal lung explants in 2% and 20% O2. Interestingly, VEGF protein was localized primarily in the basement membrane subjacent to airway epithelial cells after 4 d of incubation in 20% O2. Incubation of tissues in the presence of dibutyryl cAMP resulted in an increase in immunostaining for VEGF, primarily in the basement membranes of prealveolar ducts in 20% O2-treated tissues. In situ hybridization studies indicated that VEGF mRNA was present in both mesenchymal cells and airway epithelial cells. These data suggest that VEGF gene expression is regulated by both oxygen and cAMP in the developing human lung. The detection of VEGF mRNA and protein in distal airway epithelial cells and the detection of VEGF protein in the basement membrane subjacent to the airway epithelial cells suggest that translocation of VEGF protein occurs after its synthesis in the epithelium. Localization of VEGF to the basement membrane of airway epithelial cells may be important for directing capillary development in the human lung. PMID- 9870914 TI - Relationship between perlecan and tropoelastin gene expression and cell replication in the developing rat pulmonary vasculature. AB - Smooth-muscle-cell (SMC) replication and extracellular matrix protein expression are two vital and interrelated processes necessary for normal development of the vasculature. To understand better the nature of this relationship in the developing rat lung, we investigated the relationship between SMC proliferation and the expression of perlecan, a basement membrane (BM) heparan sulfate proteoglycan implicated in the control of SMC growth and differentiation, and tropoelastin (TE), a structural matrix protein not known to influence directly the replicative state of SMCs. Using bromodeoxyuridine (BrdU) incorporation to assess DNA synthesis, we first established the time course of SMC proliferation in the hilar pulmonary artery (PA) from embryonic to adult life. We found a labeling index of > 80% during the embryonic period (embryonic Day 13 [e13] to fetal Day 18 [f18]), a dramatic decline to approximately 40% during the fetal period of development, and a steady decrease in proliferation rates following birth such that, by 30 d of age, a labeling index of < 2% was noted. Using in situ hybridization, we found that although peak expression of both perlecan and TE messenger RNA (mRNA) occurred in the fetal and early postnatal periods following the major decrease in cell replication, TE mRNA expression was clearly observed in the PA as early as embryonic Day 14, whereas perlecan transcripts were virtually undetectable until fetal Day 19. Therefore, to evaluate further the relationship between cell replication and perlecan and/or TE gene expression, we used a combined in situ hybridization/BrdU immunohistochemistry technique and demonstrated that, on an individual cell basis, perlecan message was predominantly expressed by nonreplicating (BrdU-negative) PA, whereas TE mRNA was equally expressed in replicating and nonreplicating PA SMCs. Interestingly, a very similar pattern of replication and relationship to perlecan and TE mRNA expression was noted in airway SMCs and epithelial cells. Thus, in the lung as a whole, maximal expression of both the BM protein perlecan and the interstitial matrix protein TE occurs coordinately and follows the period of maximal SMC proliferation. However, in individual SMCs, perlecan mRNA expression varies inversely with DNA synthesis, whereas TE mRNA expression appears independent of the proliferative state of the cell. PMID- 9870915 TI - Regulation of the G protein Galphai2 by growth and development in fetal airway epithelium. AB - Heterotrimeric guanine nucleotide-binding (G) proteins transduce a wide variety of receptor-mediated signals to effectors that are involved in numerous cellular functions, including cell proliferation and differentiation. Thrombin and bombesin/gastrin-releasing peptide mediate their effects via G protein-coupled receptors to regulate lung growth and development. The growth responses of these ligands are likely to be mediated via the Gi subfamily of G proteins, specifically via Galphai2. We hypothesized that Galphai2 is expressed in the lung during ontogeny in a growth-dependent manner, and that Galphai2 regulates cell growth. We demonstrate that Galphai2 is present in the developing lung of Sprague Dawley rats, and that its expression is enhanced between embryonic Day 19 and postnatal Day 2. The strongest expression occurs in the fetal airway epithelium, and this expression in fetal airway cells is growth-dependent. Galphai2 is localized to the plasma membrane, a location consistent with interaction with growth factor receptors. Inhibition of Gi-family signal transduction by pertussis toxin (10 ng/ml) inhibits DNA synthesis in embryonic Day 19 in fetal airway epithelium. Galphai2 is likely to be a key mediator of growth signals in the developing lung. PMID- 9870916 TI - Restoration of the mucous phenotype by retinoic acid in retinoid-deficient human bronchial cell cultures: changes in mucin gene expression. AB - Retinoid-deficient cultures of airway epithelial cells undergo squamous differentiation. Treatment of such cultures with retinoic acid (RA) leads to restoration of the mucous phenotype. The purpose of our study was to characterize the cellular and molecular changes following RA treatment of retinoid-deficient human tracheobronchial epithelial cell cultures. Of particular interest was to determine when during the conversion of the squamous to the mucous phenotype the mucin genes MUC2, MUC5AC, and MUC5B were expressed. We used cornifin alpha and secreted mucin as markers to monitor the squamous and mucous phenotypes, respectively. Our studies showed that the RA responsiveness of the cultures progressively decreased with protracted retinoid deficiency, requiring higher RA concentrations to restore the mucous phenotype. Within 12 h after the start of RA treatment, cornifin alpha expression decreased, signaling the beginning of a change in cellular phenotype. At 24 h after addition of RA to the cultures, a significant number of mucous cells appeared, and at 72 h mucin was secreted in measurable amounts. Induction of mucin gene expression occurred sequentially: MUC2, MUC5AC, and MUC5B mRNAs were upregulated at 24, 48, and 72 h, respectively. When cultures maintained in 10(-8) M RA were treated with 10(-6) M RA, MUC2 but not MUC5AC and MUC5B mRNA levels were upregulated within 6 h. Our study indicates that MUC2 mRNA is an early marker of mucous differentiation, whereas MUC5AC and MUC5B mRNAs are expressed during more advanced stages of mucous differentiation. Our studies further suggest that each of the mucin genes is regulated by distinct mechanisms. PMID- 9870917 TI - The involvement of Fas-Fas ligand pathway in fibrosing lung diseases. AB - Pulmonary fibrosis begins with alveolitis, which progresses to destruction of lung tissue and excess collagen deposition. This process could be the result of DNA damage and a form of apoptosis. Therefore, we hypothesized that Fas ligand (FasL), which induces apoptosis in cells expressing Fas antigen (Fas), is associated with pulmonary fibrosis. We examined frozen lung tissues from seven patients with idiopathic pulmonary fibrosis (IPF), and bronchoalveolar lavage fluid (BALF) cells from 19 patients with IPF and from 17 patients with interstitial pneumonia associated with collagen vascular diseases (CVD-IP). We used five frozen lungs with normal lung parenchyma and BALF cells from 10 patients with solitary pulmonary nodule as controls. Reverse transcription polymerase chain reaction (RT-PCR) showed that FasL messenger RNA (mRNA) was expressed in BALF cells from all patients with IPF and from 15 of 16 patients with CVD-IP. FasL mRNA was not detected in BALF cells except in one of 10 controls. RT in situ PCR detected FasL mRNA in inflammatory cells in BALF from patients with IPF. Immunohistochemistry detected FasL protein in infiltrating lymphocytes and granulocytes in all of seven frozen lung tissues of IPF, but in none of five control lung tissues. Additionally, the expression of Fas appeared to be upregulated in bronchiolar and alveolar epithelial cells in IPF compared with normal lung parenchyma by immunohistochemistry. We conclude that Fas and FasL were upregulated in fibrosing lung diseases and may associate with DNA damage or apoptosis of bronchiolar and alveolar epithelial cells in this disorder. PMID- 9870918 TI - Interleukin-4 enhances 15-lipoxygenase activity and incorporation of 15(S)-HETE into cellular phospholipids in cultured pulmonary epithelial cells. AB - 15(S)-Hydroxyeicosatetraenoic acid (15[S]-HETE) is a 15-lipoxygenase (15-LO) metabolite that may play an important role in different pulmonary diseases. 15 HETE is synthesized by different epithelial cells and may be subsequently incorporated into cellular phospholipids. We studied the role of interleukin-4 (IL-4) on 15-LO activity and on 15(S)-HETE incorporation into cellular phospholipids by WI-26 pulmonary epithelial cells. 15-LO activity was evaluated by measuring 15(S)-HETE production, through combined reverse-phase-high-pressure liquid chromatography (RP-HPLC) separation and specific radioimmunoassay (RIA), after incubation with arachidonic acid (AA). We also studied 15-LO messenger RNA (mRNA) expression, using primed in situ (PRINS) labeling. IL-4 (10 ng/ml) markedly increased the percentage of 15-LO mRNA-bearing cells as well as 15-LO activity after 24, 48, and 72 h, with a maximal response at 48 h. Uptake and incorporation into cellular phospholipid was studied with [3H]15(S)-HETE, which showed that IL-4 was able to increase significantly 15(S)-HETE incorporation into WI-26 cells, with a maximal effect observed at 72 h. Cellular-lipid-associated [3H]15(S)-HETE, evaluated with RP-HPLC after base-catalyzed hydrolysis, increased concomitantly with disappearance of the radiolabel from the supernatant. Class separation of cellular lipids with normal-phase HPLC (NP-HPLC) showed that IL-4 increased [3H]15(S)- HETE incorporation mainly in the phosphatidylinositol (PI) fraction. The ability of IL-4 to promote 15-LO activity and incorporation into cellular phospholipids of human lung epithelial cells may be important in airway inflammation and in modulation of the potential autocrine function of 15(S)-HETE. PMID- 9870919 TI - Recombinant human Monocyte/Neutrophil elastase inhibitor protects rat lungs against injury from cystic fibrosis airway secretions. AB - Human monocyte/neutrophil elastase inhibitor (M/NEI) is a fast-acting stoichiometric inhibitor of neutrophil elastase (NE), cathepsin-G, and proteinase 3. Recombinant M/NEI (rM/NEI) was evaluated with a rat model of NE-induced lung damage. rM/NEI was found to protect against pulmonary injury caused by instilled human NE or by a preparation from airway secretions (sputum) of cystic fibrosis patients (CF sol). Human NE instilled into rat lungs produced dose-dependent hemorrhage and increased epithelial permeability, whereas NE incubated in vitro with rM/NEI did neither. Similarly, hemorrhage was induced by CF sol, but not by CF sol incubated in vitro with rM/NEI. To examine its distribution and survival time in airways, rM/NEI was labeled with the fluorochrome Texas Red (rM/NEI-TR) and instilled into rat lungs. Confocal microscopy showed that rM/NEI-TR could be detected on large airways (300 microm) at 5 min, 1 h, 4 h, and 24 h after instillation. Pretreating rats with rM/NEI was found to provide extended protection upon subsequent NE challenge, reducing hemorrhage by 98, 96, and 73%, respectively, at 1, 4, and 24 h after rM/NEI pretreatment. Pretreating rats with rM/NEI similarly conferred protection against subsequent exposure to CF sol, reducing hemorrhage by 95, 86, and 87%, respectively, at 1, 4, and 24 h after pretreatment. The findings that rM/NEI (1) mitigates protease-induced lung injury and (2) remains present and active in the lungs for 24 h after instillation strongly support its potential for treating patients with neutrophil protease induced inflammatory lung damage, such as occurs in CF and other diseases. PMID- 9870920 TI - Rab3D, a small GTPase, is localized on mast cell secretory granules and translocates to the plasma membrane upon exocytosis. AB - Although mast cell secretion has been intensively studied because of its pivotal role in allergic reactions and its advantages as a physiologic model, the molecular composition of the secretory machine is virtually unknown. In view of the guanine-nucleotide dependency of mast cell exocytosis and the participation of Rab3 proteins in synaptic vesicle release, we hypothesized that a Rab3 isoform regulates mast cell secretion. Fragments of Rab3A, 3B, and 3D were cloned from RBL-2H3 mast cells by reverse transcription- polymerase chain reaction (RT-PCR). Northern blot analysis revealed Rab3D transcripts to be relatively abundant, Rab3B substantially less so, and Rab3A and 3C undetectable. By ribonuclease (RNase) protection assay, Rab3D transcripts were at least 10-fold more abundant than those of other isoforms, and by immunoblot analysis, Rab3D protein was at least 60-fold more abundant than that of Rab3B. Rab3D was more abundant in RBL cells than in brain, but the total mass of Rab3 proteins in RBL cells was 10-fold less than in brain. Rab3D only partly colocalized with secretory granules in RBL cells, but fully colocalized in mature peritoneal mast cells. There was a descending concentration gradient of Rab3D from peripheral to central granules, and no cytoplasmic pool was detectable in resting mast cells. Following exocytotic degranulation, Rab3D translocated to the plasma membrane and remained there for at least 15 min. These studies suggest that Rab3D is a component of the regulated exocytotic machine of mast cells, and identify differences between mast cells and neurons in Rab3 expression and trafficking. PMID- 9870921 TI - Deficient hydrophilic lung surfactant proteins A and D with normal surfactant phospholipid molecular species in cystic fibrosis. AB - Chronic bacterial colonization of the lungs, with an excessive inflammatory response, is the major cause of morbidity and mortality in cystic fibrosis. Lung surfactant exhibits a spectrum of potential immunomodulatory properties: phospholipid components inhibit cellular inflammatory responses, whereas the hydrophilic surfactant proteins A (SP-A) and D (SP-D) are integral components of the innate host defense response of the lungs against bacterial infection. Consequently, alteration to the relative proportions of lung surfactant components may alter the susceptibility of the lungs to bacterial colonization. In this study, bronchoalveolar lavage (BAL) samples were collected at diagnostic fiberoptic bronchoscopy from 11 control children, 13 children with cystic fibrosis, and 11 children with acute lung infection. Electrospray ionization mass spectrometry analysis demonstrated negligible changes to the molecular species or total BAL concentrations of phosphatidylcholine, phosphatidylglycerol, or phosphatidylinositol among the three subject groups. In contrast, median SP-A concentration was decreased (P < 0.001) in the cystic fibrosis group (2.65 microg/ml) compared with control (12.35 microg/ml) and infection (9.76 microg/ml) groups. Median SP-D was also decreased (P < 0.05) in the infection (12.17 ng/ml) compared with the control group (641 ng/ml), and was below assay limits for the majority of cystic fibrosis children (P < 0. 001). This dramatic decrease of hydrophilic surfactant proteins in the presence of normal surfactant phospholipid may be one mechanism underlying the relative ineffectiveness of the cellular inflammatory response in killing invading bacteria in the lungs of patients with cystic fibrosis. PMID- 9870923 TI - Fibronectin gene polymorphisms associated with fibrosing alveolitis in systemic sclerosis. AB - Systemic sclerosis (SSc), a multisystem immunologic disease of unknown etiology, is commonly manifested in the lung as fibrosing alveolitis (FASSc). There is evidence to support the role of genetic factors in the predisposition to pulmonary fibrosis in SSc (HLA DR3/DR52a). This association is not complete and other candidate genes are likely involved. Of these, fibronectin is a growth factor known to play a crucial role in lung fibrosis. Our study investigated whether polymorphisms of the fibronectin gene are associated with lung fibrosis in SSc. Using the polymerase chain reaction and the restriction enzymes HaeIII, MspI, HindIII, and TaqI, we assessed the restriction fragment length polymorphisms (RFLPs) in 161 patients with SSc and 253 healthy control subjects from the United Kingdom. For each restriction enzyme, three genotypes were possible corresponding to the presence of the cutting site on neither, one, or both chromosomes (HaeIII AA, AB, BB; MspI CC, CD, DD; HindIII EE, EF, FF; TaqI GG, GH, HH). There was a significant decrease of genotype BB (FASSc: 17%, control: 34%; Pcorr = 0.006) with a reciprocal increase of genotype AB (FASSc: 62%, control: 46%; Pcorr = 0.022) in FASSc with the HaeIII RFLP. A significant decrease of genotype DD was observed in FASSc (FASSc: 28%, control: 41%; Pcorr = 0.038) with the MspI RFLP. The coassociation of genotypes AB (HaeIII RFLP) and CD (MspI RFLP) was present in 45% of the FASSc group (P = 0.0059), with an increased relative risk of developing fibrosing alveolitis of 1.988. We conclude that genotypes of the fibronectin gene are useful prognostic factors in SSc, helping to predict individuals likely to develop pulmonary fibrosis. PMID- 9870922 TI - Endothelin-stimulated ERK activation in airway smooth-muscle cells requires calcium influx and Raf activation. AB - Endothelin (ET)-1 is a 21-amino-acid peptide that is a potent vasoconstrictor and mitogen. By binding to its G-protein coupled receptor, ET-1 stimulates the proliferation of airway smooth-muscle (ASM) cells, which may be involved in the pathogenesis of asthma. The ETB receptor stimulates activation of the extracellular regulated kinase 2 (ERK2), which is thought to be required for proliferation of ASM cells. Our findings reveal that ET rapidly activates Raf, and that dominant-negative Raf interferes with ET-induced ERK activation in ASM cells. Expression of the amino-terminal Ras-binding domain of Raf inhibited ET induced ERK activation, suggesting that ET-stimulated Raf activation is a Ras dependent process. Furthermore, ET-stimulated ERK and Raf activation in ASM cells require calcium influx; chelating extracellular calcium or preventing calcium influx through calcium channels inhibited ET-stimulated, but not phorbol ester stimulated, ERK and Raf activation. PMID- 9870924 TI - Developmental changes in prostacyclin synthesis are conserved in cultured pulmonary endothelium and vascular smooth muscle. AB - Prostacyclin (PGI2) is a key mediator of pulmonary vascular and parenchymal function during late fetal and early postnatal life, and its synthesis in intrapulmonary arteries increases markedly during that period. The rate-limiting enzyme in PGI2 synthesis in the developing lung is cyclooxygenase (COX). To understand better the mechanisms underlying the developmental increase in PGI2 synthesis, we evaluated PGI2 production in early-passage, cultured pulmonary artery endothelial cells (PAEC) and pulmonary vascular smooth-muscle cells (VSM) from fetal and newborn lambs. In arterial segments, PGI2 synthesis was sevenfold greater in intact arteries from newborn than from fetal lambs, and it was 12-fold greater in endothelium-denuded newborn than in fetal arteries, indicating that the developmental increase occurs in both the endothelium and medial layer. Similarly, basal PGI2 production was three-fold greater in newborn than in fetal PAEC, and 2.5-fold greater in newborn than in fetal pulmonary VSM cells. Calcium ionophore (A23187)-stimulated and arachidonic acid-stimulated PGI2 synthesis were also greater in newborn than in fetal PAEC and VSM, revealing a developmental upregulation in COX enzymatic activity in both cell types. Immunoblot analysis showed that this is due to greater COX-1 protein expression in newborn than in fetal vascular cells; COX-2 protein expression was not detected. In addition, COX 1 messenger RNA (mRNA) abundance was greater in newborn than in fetal PAEC, and this was not due to a difference in COX-1 mRNA stability. Thus, the developmental upregulation of PGI2 synthesis is conserved in early-passage PAEC and pulmonary VSM, and is related to a maturational increase in COX-1 gene expression. Further studies with the cultured cell model will enable determination of the factors that directly regulate COX-1 expression in the developing pulmonary vasculature. PMID- 9870926 TI - The in vivo effects of milrinone on the airways of cystic fibrosis mice and human subjects. AB - Previous studies have indicated that milrinone, a specific type III phosphodiesterase inhibitor, may be able to induce chloride secretion in cystic fibrosis (CF) tissues. We have now assessed the effect of this agent in vivo on the nasal epithelium of CF mutant mice and also in the nose and lungs of human subjects with CF. Wild-type mice showed a small hyperpolarization of the nasal potential difference (PD) in response to milrinone (100 microM, 1.6 +/- 0.6 mV, n = 8, P < 0.05). In contrast, CF mice carrying either the most common human mutation of the gene for the CF transmembrane regulator (CFTR), DeltaF508 (protein mislocalized), or the G551D mutation (protein normally localized) failed to demonstrate this response. Milrinone perfused alone had no significant effect on the baseline nasal PD of human subjects without CF (14.7 +/- 4.0 mV preperfusion; 15.3 +/- 4.6 mV postperfusion), but significantly (P < 0.05) augmented the hyperpolarization induced by a subsequently perfused low-chloride solution (with milrinone, 36.8 +/- 3.0 mV, n = 6; without milrinone, 18.1 +/- 2.2 mV, n = 19). In contrast, in human subjects with CF (n = 6), milrinone alone significantly (P < 0. 05) altered the nasal baseline PD (52.2 +/- 3.3 mV preperfusion; 57. 4 +/- 4.2 mV, postperfusion) but not the subsequent responses to the low-chloride solution (with milrinone, 1.1 +/- 2.2 mV, n = 4; without milrinone, 0.6 +/- 0.5 mV, n = 28) or to isoproterenol (100 microM). In a separate study in subjects (n = 6) with the DeltaF508 mutation, nasal coadministration of milrinone with isoproterenol produced no effect in the presence of amiloride and a low-chloride solution (-0.8 +/- 0.5 mV). This was also the case in the nasal epithelium of CF subjects (n = 4) carrying at least one G551D allele (-0.3 +/- 0.8 mV). Similarly, milrinone did not hyperpolarize the PD of either the tracheal (n = 6) or segmental (n = 6) airways of CF subjects (DeltaF508) when applied topically in vivo in the presence of amiloride, isoproterenol, or adenosine triphosphate (all 100 microM) in a low-chloride solution. These data do not support the use of milrinone to induce chloride secretion in CF airways in vivo. PMID- 9870927 TI - Phenotypic and functional characterization of normal rat pleural macrophages in comparison with autologous peritoneal and alveolar macrophages. AB - Pleural mononuclear phagocytes (PleMP) were isolated from normal rats by pleural lavage and compared with autologous peritoneal (PerMP) and bronchoalveolar mononuclear phagocytes (BAMP) isolated by peritoneal and bronchoalveolar lavage, respectively. The phagocytic activity of PleMP, PerMP, and BAMP, evaluated by testing their ability to ingest latex beads, was lower for PleMP and PerMP than for BAMP. The phenotype of PleMP, PerMP, and BAMP was characterized by immunocytochemical staining with a panel of monoclonal antibodies (mAbs). As expected, PleMP, PerMP, and BAMP did not react with OX19, OX33, ED5, MOM/3F12/F2, and anticytokeratin mAbs, specific for T lymphocytes, B lymphocytes, dendritic cells, granulocytes, and epithelial/mesothelial cells, respectively. Moreover, PleMP and PerMP populations were highly enriched with OX6-, OX42-, ED7-, and ED8 positive MP, whereas BAMP population was enriched with ED1- and ED9-positive cells. To test the ability of PleMP, PerMP, and BAMP to function as accessory cells (AC), mitomycin C-treated MP were used as stimulatory cells in mixed leukocyte reaction experiments, using allogeneic T cells as responders. 3HdTR incorporation by T cells was assessed as an index of AC function. PleMP and PerMP were more potent AC than BAMP. Moreover, when cultured together with autologous pulmonary interstitial dendritic cells, PleMP and PerMP exerted a more potent ability to stimulate T-cell proliferation than did BAMP. To investigate the capacity of MP to function as bactericidal and fungicidal cells, we tested their ability to kill Escherichia coli and Cryptococcus neoformans, respectively. PleMP and PerMP were less potent bactericidal and fungicidal cells than BAMP. The results of this study demonstrate that PleMP isolated from normal rat pleural space are functionally and phenotypically different from BAMP but similar to PerMP, and suggest that these cells might play an important role in cell-mediated immune reactions in the pleural space. PMID- 9870925 TI - Hydrogen peroxide enhances shedding of type I soluble tumor necrosis factor receptor from pulmonary epithelial cells. AB - Reactive oxygen intermediates (ROIs) are among the important mediators in the pathogenesis of lung diseases in which tumor necrosis factor (TNF) plays a pivotal role. However, the effects of ROIs on the TNF- TNF receptor system remain unclear. Effects of hydrogen peroxide on the shedding of soluble tumor necrosis factor receptor (sTNF-R) were investigated in a pulmonary epithelial cell line (A549) using enzyme-linked immunoassay. A549 cells spontaneously released type I sTNF-R (sTNF-RI) into the culture medium. Hydrogen peroxide accelerated the release of sTNF-RI from the A549 cells time- and dose- dependently. Stimulated release of sTNF-RI by hydrogen peroxide or phorbol myristate acetate (PMA) was inhibited by pretreatment with the intracellular hydroxyl radical scavengers dimethyl sulfoxide and dimethyl thiourea. A synthetic metalloproteinase inhibitor (KB-R8301) inhibited not only spontaneous release of sTNF-RI but also shedding enhanced by hydrogen peroxide and PMA. Preincubation with a protein kinase C inhibitor, calphostin C, downregulated the hydrogen peroxide- or PMA-induced shedding of sTNF-RI. Neither genistein, a tyrosine kinase inhibitor, nor H-89, a protein kinase A inhibitor, inhibited shedding of sTNF-RI by hydrogen peroxide and PMA. Although the surface expression of TNF-R assessed by 125I-TNF specific binding was decreased in the presence of hydrogen peroxide or PMA, TNF-RI mRNA transcript levels remained unchanged. These results show that hydrogen peroxide is involved in the activation of metalloproteinase and protein kinase C responsible for the shedding of sTNF-RI. Accordingly, ROIs may alter TNF action by enhanced shedding of sTNF-RI and reducing its surface receptor expression. PMID- 9870928 TI - Expression and inducibility of alpha, pi, and Mu glutathione S-transferase protein and mRNA in murine lung. AB - This investigation sought to establish the cellular expression and distribution of the alpha, pi, and mu classes of glutathione S-transferase (GST) enzymes in murine lung under control conditions and after treatment with tert-butyl-4 hydroxyanisole (BHA). Immunohistochemical and in situ hybridization studies were used to identify lung cells that were labeled for the GST subunits Yp, Ya, and Yb1. Immunoblotting of cytosolic proteins produced single bands of 28, 29, and 31 kD for Ya, Yp, and Yb1, respectively, in samples from untreated and BHA-treated mice. Treatment with BHA increased Ya and Yp reactivity, but this was not as marked for Yb1. Immunohistochemical staining for the Yp, Ya, and Yb1 subunits was localized in bronchioles and parenchyma of untreated and BHA-treated mice. Bronchiolar Clara and alveolar type II cells were stained to the greatest extent for all of the GST subunits. BHA treatment produced increased staining that was most pronounced in the bronchiolar epithelium. Ya and Yp were localized in the cytoplasm and nucleus, whereas Yb1 was found mainly in the cytoplasm. Immunoblots of extracted nuclear proteins revealed a band of 29 kD for Ya, with increased immunoreactivity in BHA-treated mice. In situ hybridization done with oligonucleotide probes showed abundant silver grains representing Ya, Yp, and Yb1 messenger RNA (mRNA) transcripts in the bronchioles. Grains were also localized in alveolar septa, and were most numerous in type II cells. Quantitative image analysis confirmed good agreement between relative levels of protein and mRNA transcripts. Quantities of mRNA transcripts for all subunits were increased in the parenchyma by BHA treatment, but the magnitudes of induction were most striking for Ya and Yp in the bronchioles. These results demonstrated that Ya, Yp, and Yb1 reside in specific lung areas and cells, and that in induced states, their increased expression is accompanied by increased mRNA. PMID- 9870929 TI - B7 costimulation is required for IL-5 and IL-13 secretion by bronchial biopsy tissue of atopic asthmatic subjects in response to allergen stimulation. AB - Asthma is a complex disorder characterized by airway hyperreactivity and inflammation. To analyze cellular interactions required for the secretion of cytokines by the bronchial mucosa, we have evaluated the ex vivo response of tissue explants to allergen. Endobronchial mucosal biopsy tissue from mild atopic asthmatic subjects and normal control subjects were maintained in culture for 24 h. To detect reactivity to allergen, the explants were stimulated with dust mite extract Dermatophagoides pteronyssinus (Der p). Our analysis revealed that without any overt stimulation, mRNA transcripts for interleukin (IL)-5 and IL-13 were expressed by asthmatic but not normal bronchial tissue. In contrast, the expression of interferon-gamma was observed in a higher proportion of cultured bronchial biopsies from the normal control subjects than in those from asthmatic subjects. Addition of Der p allergen did not change the cytokine profile of the explants from control volunteers but augmented the expression of IL-5 mRNA and induced secretion of the protein by the asthmatic bronchial tissue. In most cases, allergen also increased the production of IL-13 by bronchial tissue from asthmatic subjects. The allergen-induced secretion of IL-5 and IL-13 was inhibited by the fusion protein CTLA-4Ig, reflecting a requirement for CD80 (B7 1) and/or CD86 (B7-2) costimulation for the expression of the Th2 cytokines. This requirement for B7/CD28 costimulation is consistent with the hypothesis that IL-5 and IL-13 are produced by allergen-specific T cells resident in the asthmatic bronchial mucosa. PMID- 9870930 TI - 5-oxo-6,8,11,14-eicosatetraenoic acid is a potent stimulator of L-selectin shedding, surface expression of CD11b, actin polymerization, and calcium mobilization in human eosinophils. AB - 5-oxo-6,8,11,14-eicosatetraenoic acid (5-oxo-ETE) is a metabolite of arachidonic acid formed by the oxidation of 5-hydroxy-6,8,11, 14-eicosatetraenoic acid by a highly specific dehydrogenase. 5-oxo-ETE is a chemoattractant for both neutrophils and eosinophils. Although it is not as effective as leukotriene B4 (LTB4) and platelet-activating factor (PAF) in stimulating neutrophil migration, we found that it is considerably more active than these and a variety of other lipid mediators as an eosinophil chemoattractant. Moreover, low concentrations of 5-oxo-ETE appear to enhance the responsiveness of these cells to PAF. The objectives of the current investigation were to identify rapid responses induced in eosinophils by 5-oxo-ETE that might be related to the infiltration of these cells into tissues. We found that 5-oxo-ETE is more effective than PAF and LTB4 in inducing both L-selectin shedding and actin polymerization in human eosinophils, whereas PAF is the most active of these mediators in stimulating calcium mobilization. The complementary effects of 5-oxo-ETE and PAF on actin polymerization and calcium mobilization may explain their synergistic effect on eosinophil migration. 5-oxo-ETE and PAF were equipotent in stimulating the surface expression of the beta2-integrin CD11b, but were slightly less potent than LTB4. 5-oxo-ETE- induced actin polymerization was subject to homologous but not heterologous desensitization. It was not prevented by incubation of eosinophils with inhibitors of protein kinase C (staurosporine), mitogen activated protein kinase kinase (PD98059), or phosphatidylinositol-3-kinase (wortmannin). In conclusion, 5-oxo-ETE is a potent activator of human eosinophils and may be an important regulator of tissue infiltration of these cells. PMID- 9870931 TI - Cigarette smoking causes sequestration of polymorphonuclear leukocytes released from the bone marrow in lung microvessels. AB - Studies from our laboratory have shown that chronic cigarette smoke exposure causes a neutrophilia associated with a shortening of the mean transit time of polymorphonuclear leukocytes (PMN) though the postmitotic pool of the marrow. The present study was designed to test the hypothesis that PMN newly released from bone marrow by smoke exposure preferentially sequestered in pulmonary microvessels. The thymidine analogue 5'-bromo-2'-deoxyuridine (BrdU) was used to label dividing PMN in the marrow of rabbits; their appearance in the circulation was measured using immunocytochemistry, and their sequestration in lung tissue was determined using standard morphometric techniques. Animals exposed to 11 d of cigarette smoke (n = 6) compared with sham-exposed control animals (n = 4) showed no increase in circulating PMN counts but showed an increase in both the percentage of band cells (smoking, 9.8 +/- 1.1% versus control, 5.5 +/- 0.9%; P < 0.05) and BrdU-labeled PMN (PMNBrdU) in the circulation (smoking, 10.8 +/- 0.6% versus control, 7.5 +/- 0.3%; P < 0.05). There were more PMN sequestered in the lungs of smoke-exposed animals (51.7 +/- 3.4 x 10(7)/ml tissue) than in those of control animals (25.1 +/- 1.8 x 10(7)/ ml tissue) (P < 0.05) and a higher percentage of these cells were PMNBrdU (smoking, 16.9 +/- 2. 3% versus control, 9.6 +/- 0.4%; P < 0.05). The percentage of PMNBrdU in the gravity-independent regions (11.7 +/- 1.9%) of the lung was higher than gravity-dependent regions (7.8 +/- 1.8%) in the smoke-exposure group (P < 0.05). Transmission electron microscopy showed pulmonary capillary endothelial damage with adherent PMN in the smoke-exposure group. We conclude that younger PMN released from the bone marrow by cigarette smoking preferentially sequestered in pulmonary microvessels and speculate that these PMN may contribute to the alveolar wall damage associated with smoke-induced lung emphysema. PMID- 9870932 TI - Synthesis of beta-tubulin, actin, and other proteins in axons of sympathetic neurons in compartmented cultures. AB - The proteins needed for growth and maintenance of the axon are generally believed to be synthesized in the cell bodies and delivered to the axons by anterograde transport. However, recent reports suggest that some proteins can also be synthesized within axons. We used [35S]methionine metabolic labeling to investigate axonal protein synthesis in compartmented cultures of sympathetic neurons from newborn rats. Incubation of distal axons for 4 hr with [35S]methionine resulted in a highly specific pattern of labeled axonal proteins on SDS-PAGE, with 4 prominent bands in the 43-55 kDa range. The labeled proteins in axons were not synthesized in the cell bodies, because they were also produced by axons after the cell bodies had been removed. Two of the proteins were identified by immunoprecipitation as actin and beta-tubulin. Axons synthesized <1% of the actin and tubulin synthesized in the cell bodies and transported into the axons, and 75-85% inhibition of axonal protein synthesis by cycloheximide and puromycin failed to inhibit axonal elongation. Nonetheless, the specific production by axons of the major proteins of the axonal cytoskeleton suggests that axonal protein synthesis arises from specific mechanisms and likely has biological significance. One hypothetical scenario involves neurons with long axons in vivo in which losses from turnover during axonal transport may limit the availability of cell body synthesized proteins to the distal axons. In this case, a significant fraction of axonal proteins might be supplied by axonal synthesis, which could, therefore, play important roles in axonal maintenance, regeneration, and sprouting. PMID- 9870934 TI - The mammalian brain high-affinity L-proline transporter is enriched preferentially in synaptic vesicles in a subpopulation of excitatory nerve terminals in rat forebrain. AB - The expression of a brain-specific high-affinity Na+-dependent (and Cl- dependent) L-proline transporter (PROT) in subpopulations of putative glutamatergic neurons in mammalian brain suggests a physiological role for this carrier in excitatory neurotransmission (). To gain insights into potential sites where PROT may function, we used a C-terminal domain antipeptide antibody to determine the regional distribution and subcellular localization of PROT in rat forebrain. PROT immunoreactivity was seen in processes having a regional light microscopic distribution comparable to that of known glutamatergic projections within the cortex, caudate putamen nucleus (CPN), hippocampal formation, and other forebrain regions. In all regions examined by electron microscopy (cortex, CPN, and the stratum oriens of CA1), PROT labeling was observed primarily within subpopulations of axon terminals forming asymmetric excitatory-type synapses. Immunogold labeling for PROT was detected in close contact with membranes of small synaptic vesicles (SSVs) and more rarely with the plasma membrane in these axon terminals. Subcellular fractionation studies confirmed the preferential distribution of PROT to synaptic vesicles. The topology of PROT in synaptic vesicles was found to be inverted with respect to the plasma membrane, suggesting that PROT-containing vesicles are generated by a process involving endocytosis from the plasma membrane. Because PROT lacks any of the known characteristics of other vesicular transporters, these results suggest that certain excitatory terminals have a reserve pool of PROT associated with SSVs. The delivery of PROT to the plasma membrane by exocytosis could play a critical role in the plasticity of certain glutamatergic pathways. PMID- 9870933 TI - Neuroprotection and neuronal differentiation studies using substantia nigra dopaminergic cells derived from transgenic mouse embryos. AB - The major pathological lesion of Parkinson's disease (PD) is the selective cell death of dopaminergic (DA) neurons in substantia nigra (SN). Although the initial cause and subsequent molecular signaling mechanisms leading to DA cell death underlying the PD process remain elusive, brain-derived neurotrophic factor (BDNF) is thought to exert neuroprotective as well as neurotrophic roles for the survival and differentiation of DA neurons in SN. Addressing molecular mechanisms of BDNF action in both primary embryonic mesencephalic cultures and in vivo animal models has been technically difficult because DA neurons in SN are relatively rare and present with many heterogeneous cell populations in midbrain. We have developed and characterized a DA neuronal cell line of embryonic SN origin that is more accessible to molecular analysis and can be used as an in vitro model system for studying SN DA neurons. A clonal SN DA neuronal progenitor cell line SN4741, arrested at an early DA developmental stage, was established from transgenic mouse embryos containing the targeted expression of the thermolabile SV40Tag in SN DA neurons. The phenotypic and morphological differentiation of the SN4741 cells could be manipulated by environmental cues in vitro. Exogenous BDNF treatment produced significant neuroprotection against 1 methyl-4-phenylpyridinium, glutamate, and nitric oxide-induced neurotoxicity in the SN4741 cells. Simultaneous phosphorylation of receptor tyrosine kinase B accompanied the neuroprotection. This SN DA neuronal cell line provides a unique model system to circumvent the limitations associated with primary mesencephalic cultures for the elucidation of molecular mechanisms of BDNF action on DA neurons of the SN. PMID- 9870935 TI - An increase in lactate output by brain tissue serves to meet the energy needs of glutamate-activated neurons. AB - Aerobic energy metabolism uses glucose and oxygen to produce all the energy needs of the brain. Several studies published over the last 13 years challenged the assumption that the activated brain increases its oxidative glucose metabolism to meet the increased energy demands. Neuronal function in rat hippocampal slices supplied with 4 mM glucose could tolerate a 15 min activation by a 5 mM concentration of the excitatory neurotransmitter glutamate (Glu), whereas slices supplied with 10 mM glucose could tolerate a 15 min activation by 20 mM Glu. However, in slices in which neuronal lactate use was inhibited by the lactate transporter inhibitor a-cyano-4-hydroxycinnamate (4-CIN), activation by Glu elicited a permanent loss of neuronal function, with a twofold to threefold increase in tissue lactate content. Inhibition of glycolysis with the glucose analog 2-deoxy-D-glucose (2DG) during the period of exposure to Glu diminished normal neuronal function in the majority of slices and significantly reduced the number of slices that exhibited neuronal function after activation. However, when lactate was added with 2DG, the majority of the slices were neuronally functional after activation by Glu. NMDA, a nontransportable Glu analog by the glial glutamate transporter, could not induce a significant increase in slice lactate level when administered in the presence of 4-CIN. It is suggested that the heightened energy demands of activated neurons are met through increased glial glycolytic flux. The lactate thus formed is a crucial aerobic energy substrate that enables neurons to endure activation. PMID- 9870936 TI - A mathematical model for the intracellular circadian rhythm generator. AB - A mathematical model for the intracellular circadian rhythm generator has been studied, based on a negative feedback of protein products on the transcription rate of their genes. The study is an attempt at examining minimal but biologically realistic requirements for a negative molecular feedback loop involving considerably faster reactions, to produce (slow) circadian oscillations. The model included mRNA and protein production and degradation, along with a negative feedback of the proteins upon mRNA production. The protein production process was described solely by its total duration and a nonlinear term, whereas also the feedback included nonlinear interactions among protein molecules. This system was found to produce robust oscillations in protein and mRNA levels over a wide range of parameter values. Oscillations were slow, with periods much longer than the time constants of any of the individual system parameters. Circadian oscillations were obtained for realistic values of the parameters. The system was readily entrainable to external periodic perturbations. Two distinct classes of phase response curves were found, viz. with or without a time domain within the circadian cycle in which external perturbations fail to induce a phase shift ("dead zone"). The delay and nonlinearity in the protein production and the cooperativity in the negative feedback (Hill coefficient) were for this model found to be necessary and sufficient to generate robust circadian oscillations. The similarities between model outcomes and empirical findings establish that circadian rhythmicity at the cellular level can plausibly emerge from interactions among molecular systems which are not in themselves rhythmic. PMID- 9870937 TI - Long-term potentiation in the hippocampal CA1 region of mice lacking cGMP dependent kinases is normal and susceptible to inhibition of nitric oxide synthase. AB - Long-term potentiation (LTP) is a potential cellular mechanism for learning and memory. The retrograde messenger nitric oxide (NO) is thought to induce LTP in the CA1 region of the hippocampus via activation of soluble guanylyl cyclase (sGC) and, ultimately, cGMP-dependent protein kinase (cGK). Two genes code for the isozymes cGKI and cGKII in vertebrates. The functional role of cGKs in LTP was analyzed using mice lacking the gene(s) for cGKI, cGKII, or both. LTP was not altered in the mutant mice lineages. However, LTP was reduced by inhibition of NO synthase and NMDA receptor antagonists, respectively. The reduced LTP was not recovered by the cGK-activator 8-(4 chlorophenylthio)-cGMP. Moreover, LTP was not affected by the sGC inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]-quiloxalin-1-one. In contrast, it was effectively suppressed by nicotinamide, a blocker of the ADP ribosyltransferase. These results show that cGKs are not involved in LTP in mice and that NO induces LTP through an alternative cGMP-independent pathway, possibly ADP-ribosylation. PMID- 9870938 TI - Requirement of receptor internalization for opioid stimulation of mitogen activated protein kinase: biochemical and immunofluorescence confocal microscopic evidence. AB - Previously, we implicated the opioid receptor (OR), Gbetagamma subunits, and Ras in the opioid activation of extracellular signal-regulated protein kinase (ERK), a member of the mitogen-activated protein (MAP) kinase family involved in mitogenic signaling. We now report that OR endocytosis also plays a role in the opioid stimulation of ERK activity. COS-7 and HEK-293 cells were cotransfected with the cDNA of delta-, mu;-, or kappa-OR, dynamin wild-type (DWT), or the dominant suppressor mutant dynamin K44A, which blocks receptor endocytosis. The activation of ERK by opioid agonists in the presence of DWT was detected. In contrast, parallel ectopic coexpression of the K44A mutant with OR, followed by agonist treatment, resulted in a time-dependent attenuation of ERK activation. Immunofluorescence confocal microscopy of delta-OR and DWT-cotransfected COS-7 cells revealed that agonist exposure for 10 min resulted in an ablation of cell surface delta-OR immunoreactivity (IR) and an intensification of cytoplasmic (presumably endosomal) staining as seen in the absence of overexpressed DWT. After 1 hr of delta-agonist exposure the cells displayed substantial internalization of delta-OR IR. If the cells were cotransfected with delta-OR and dynamin mutant K44A, OR IR was retained on the cell surface even after 1 hr of delta-agonist treatment. Parallel immunofluorescence confocal microscopy, using an anti-ERK antibody, showed that agonist-induced time-dependent ERK IR trafficking into perinuclear and nuclear loci was impaired in the internalization defective cells. Thus, both biochemical and immunofluorescence confocal microscopic evidence supports the hypothesis that the opioid activation of ERK requires receptor internalization in transfected mammalian cells. PMID- 9870939 TI - Mechanisms and structural determinants of HIV-1 coat protein, gp41-induced neurotoxicity. AB - Of the individuals with human immunodeficiency virus type 1 (HIV-1) infection, 20 30% will develop the neurological complication of HIV-associated dementia (HAD). The mechanisms underlying HAD are unknown; however, indirect immunologically mediated mechanisms are theorized to play a role. Recently, the HIV-1 coat protein gp41 has been implicated as a major mediator of HAD through induction of neurocytokines and subsequent neuronal cell death. Using primary mixed cortical cultures from neuronal nitric oxide synthase (NOS) null (nNOS-/-) mice and immunological NOS null (iNOS-/-) mice, we establish iNOS-derived NO as a major mediator of gp41 neurotoxicity. Neurotoxicity elicited by gp41 is markedly attenuated in iNOS-/- cultures compared with wild-type and nNOS-/- cultures. The NOS inhibitor L-nitroarginine methyl ester is neuroprotective in wild-type and nNOS-/- cultures, confirming the role of iNOS-derived NO in gp41 neurotoxicity. Confirming that iNOS-/- cultures lack iNOS, gp41 did not induce iNOS in iNOS-/- cultures, but it markedly induced iNOS in wild-type and nNOS-/- cultures. We elucidate the region of gp41 that is critical for iNOS induction and neuronal cell death by monitoring iNOS induction with overlapping peptides spanning gp41. We show that the N-terminal region of gp41, which we designate as the neurotoxic domain, induces iNOS protein activity and iNOS-dependent neurotoxicity at picomolar concentrations in a manner similar to recombinant gp41 protein. Our experiments suggest that gp41 is eliciting the induction of iNOS through potential cell surface receptors or binding sites because the induction of iNOS is dose dependent and saturable and occurs at physiologically relevant concentrations. These data confirm that the induction of iNOS by gp41 and the production of NO are primary mediators of neuronal damage and identify a neurotoxic domain of gp41 that may play an important role in HAD. PMID- 9870940 TI - The cat-1 gene of Caenorhabditis elegans encodes a vesicular monoamine transporter required for specific monoamine-dependent behaviors. AB - We have identified the Caenorhabditis elegans homolog of the mammalian vesicular monoamine transporters (VMATs); it is 47% identical to human VMAT1 and 49% identical to human VMAT2. C. elegans VMAT is associated with synaptic vesicles in approximately 25 neurons, including all of the cells reported to contain dopamine and serotonin, plus a few others. When C. elegans VMAT is expressed in mammalian cells, it has serotonin and dopamine transport activity; norepinephrine, tyramine, octopamine, and histamine also have high affinity for the transporter. The pharmacological profile of C. elegans VMAT is closer to mammalian VMAT2 than VMAT1. The C. elegans VMAT gene is cat-1; cat-1 knock-outs are totally deficient for VMAT immunostaining and for dopamine-mediated sensory behaviors, yet they are viable and grow relatively well. The cat-1 mutant phenotypes can be rescued by C. elegans VMAT constructs and also (at least partially) by human VMAT1 or VMAT2 transgenes. It therefore appears that the function of amine neurotransmitters can be completely dependent on their loading into synaptic vesicles. PMID- 9870941 TI - Opioid receptor subtype expression defines morphologically distinct classes of hippocampal interneurons. AB - The inhibition of hippocampal pyramidal cells occurs via inhibitory interneurons making GABAergic synapses on distinct segments of the postsynaptic membrane. In area CA1 of the hippocampus, the activation of mu- and delta-opioid receptors inhibits these interneurons, thereby increasing the excitability of the pyramidal cells. Through the use of selective opioid agonists and biocytin-filled whole cell electrodes, interneurons possessing somata located within stratum oriens of hippocampal slices were classified according to the location of their primary axon termination and the expression of mu- or delta-opioid receptors. Activation of these opioid receptor subtypes resulted in outward currents in the majority of interneurons, which is consistent with their inhibition. Post hoc morphological analysis revealed that those interneurons heavily innervating the pyramidal cell body layer were much more likely to express mu-opioid receptors, whereas cells with axons ramifying in the pyramidal neuron dendritic layers were more likely to express delta-opioid receptors, as defined by the generation of outward currents. This morphological segregation of interneuron projections suggests that mu receptor activation would diminish GABA release onto pyramidal neuron somata, thereby increasing their excitability and output. Conversely, inhibition of interneurons via delta receptor activation would amplify afferent signaling to pyramidal neuron dendrites by reducing GABAergic inhibition of these structures. PMID- 9870942 TI - Citron binds to PSD-95 at glutamatergic synapses on inhibitory neurons in the hippocampus. AB - Synaptic NMDA-type glutamate receptors are anchored to the second of three PDZ (PSD-95/Discs large/ZO-1) domains in the postsynaptic density (PSD) protein PSD 95. Here, we report that citron, a protein target for the activated form of the small GTP-binding protein Rho, preferentially binds the third PDZ domain of PSD 95. In GABAergic neurons from the hippocampus, citron forms a complex with PSD-95 and is concentrated at the postsynaptic side of glutamatergic synapses. Citron is expressed only at low levels in glutamatergic neurons in the hippocampus and is not detectable at synapses onto these neurons. In contrast to citron, p135 SynGAP, an abundant synaptic Ras GTPase-activating protein that can bind to all three PDZ domains of PSD-95, and Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) are concentrated postsynaptically at glutamatergic synapses on glutamatergic neurons. CaM kinase II is not expressed and p135 SynGAP is expressed in less than half of hippocampal GABAergic neurons. Segregation of citron into inhibitory neurons does not occur in other brain regions. For example, citron is expressed at high levels in most thalamic neurons, which are primarily glutamatergic and contain CaM kinase II. In several other brain regions, citron is present in a subset of neurons that can be either GABAergic or glutamatergic and can sometimes express CaM kinase II. Thus, in the hippocampus, signal transduction complexes associated with postsynaptic NMDA receptors are different in glutamatergic and GABAergic neurons and are specialized in a way that is specific to the hippocampus. PMID- 9870943 TI - Citron, a Rho-target, interacts with PSD-95/SAP-90 at glutamatergic synapses in the thalamus. AB - Proteins of the membrane-associated guanylate kinase family play an important role in the anchoring and clustering of neurotransmitter receptors in the postsynaptic density (PSD) at many central synapses. However, relatively little is known about how these multifunctional scaffold proteins might provide a privileged site for activity- and cell type-dependent specification of the postsynaptic signaling machinery. Rho signaling pathway has classically been implicated in mechanisms of axonal outgrowth, dendrogenesis, and cell migration during neural development, but its contribution remains unclear at the synapses in the mature CNS. Here, we present evidence that Citron, a Rho-effector in the brain, is enriched in the PSD fraction and interacts with PSD-95/synapse associated protein (SAP)-90 both in vivo and in vitro. Citron colocalization with PSD-95 occurred, not exclusively but certainly, at glutamatergic synapses in a limited set of neurons, such as the thalamic excitatory neurons; Citron expression, however, could not be detected in the principal neurons of the hippocampus and the cerebellum in the adult mouse brain. In a heterologous system, Citron was shown to form a heteromeric complex not only with PSD-95 but also with NMDA receptors. Thus, Citron-PSD-95/SAP-90 interaction may provide a region- and cell type-specific link between the Rho signaling cascade and the synaptic NMDA receptor complex. PMID- 9870944 TI - Synaptic vesicle populations in saccular hair cells reconstructed by electron tomography. AB - We used electron tomography to map the three-dimensional architecture of the ribbon-class afferent synapses in frog saccular hair cells. The synaptic body (SB) at each synapse was nearly spherical (468 +/- 65 nm diameter; mean +/- SD) and was covered by a monolayer of synaptic vesicles (34.3 nm diameter; 8.8% coefficient of variation), many of them tethered to it by approximately 20-nm long filaments, at an average density of 55% of close-packed (376 +/- 133 vesicles per SB). These vesicles could support approximately 900 msec of exocytosis at the reported maximal rate, which the cells can sustain for at least 2 sec, suggesting that replenishment of vesicles on the SB is not rate limiting. Consistent with this interpretation, prolonged K+ depolarization did not deplete vesicles on the SB. The monolayer of SB-associated vesicles remained after cell lysis in the presence of 4 mM Ca2+, indicating that the association is tight and Ca2+-resistant. The space between the SB and the plasma membrane contained numerous vesicles, many of which ( approximately 32 per synapse) were in contact with the plasma membrane. This number of docked vesicles could support maximal exocytosis for at most approximately 70 msec. Additional docked vesicles were seen within a few hundred nanometers of the synapse and occasionally at greater distances. The presence of omega profiles on the plasma membrane around active zones, in the same locations as coated pits and coated vesicles labeled with an extracellular marker, suggests that local membrane recycling may contribute to the three- to 14-fold greater abundance of vesicles in the cytoplasm (not associated with the SB) near synapses than in nonsynaptic regions. PMID- 9870945 TI - Heparin-binding epidermal growth factor-like growth factor in hippocampus: modulation of expression by seizures and anti-excitotoxic action. AB - The expression of heparin-binding epidermal growth factor-like growth factor (HB EGF), an EGF receptor ligand, was investigated in rat forebrain under basal conditions and after kainate-induced excitotoxic seizures. In addition, a potential neuroprotective role for HB-EGF was assessed in hippocampal cultures. In situ hybridization analysis of HB-EGF mRNA in developing rat hippocampus revealed its expression in all principle cell layers of hippocampus from birth to postnatal day (P) 7, whereas from P14 through adulthood, expression decreased in the pyramidal cell layer versus the dentate gyrus granule cells. After kainate induced excitotoxic seizures, levels of HB-EGF mRNA increased markedly in the hippocampus, as well as in several other cortical and limbic forebrain regions. In the hippocampus, HB-EGF mRNA expression increased within 3 hr after kainate treatment, continued to increase until 24 hr, and then decreased; increases occurred in the dentate gyrus granule cells, in the molecular layer of the dentate gyrus, and in and around hippocampal pyramidal CA3 and CA1 neurons. At 48 hr after kainate treatment, HB-EGF mRNA remained elevated in vulnerable brain regions of the hippocampus and amygdaloid complex. Western blot analysis revealed increased levels of HB-EGF protein in the hippocampus after kainate administration, with a peak at 24 hr. Pretreatment of embryonic hippocampal cell cultures with HB-EGF protected neurons against kainate toxicity. The kainate induced elevation of [Ca2+]i in hippocampal neurons was not altered in cultures pretreated with HB-EGF, suggesting an excitoprotective mechanism different from that of previously characterized excitoprotective growth factors. Taken together, these results suggest that HB-EGF may function as an endogenous neuroprotective agent after seizure-induced neural activity/injury. PMID- 9870946 TI - Nitric oxide acutely inhibits neuronal energy production. The Committees on Neurobiology and Cell Physiology. AB - Disruption of mitochondrial respiration has been proposed as an action of nitric oxide (NO) responsible for its toxicity, but the effects of NO on the energetics of intact central neurons have not been reported. We examined the effects of NO on mitochondrial function and energy metabolism in cultured hippocampal neurons. The application of NO from NO donors or from dissolved gas produced a rapid, reversible depolarization of mitochondrial membrane potential, as detected by rhodamine-123 fluorescence. NO also produced a progressive concentration dependent depletion of cellular ATP over 20 min exposures. The energy depletion produced by higher levels of NO (2 microM or more) was profound and irreversible and proceeded to subsequent neuronal death. In contrast to the effects of NO, mitochondrial protonophores produced complete depolarizations of mitochondrial membrane potential but depleted the neuronal ATP stores only partially. Inhibitors of mitochondrial oxidative phosphorylation (rotenone or 3 nitropropionic acid) or of glycolysis (iodoacetate plus pyruvate) also produced only partial ATP depletion, suggesting that either process alone could partially maintain ATP stores. Only by combining the inhibition of glycolytic energy production with the inhibition of mitochondria could the effects of NO in depleting energy and inducing delayed toxicity be duplicated. These results show that NO has rapid inhibitory actions on mitochondrial metabolism in living neurons. However, the severe ATP-depleting effects of high concentrations of NO are not fully explained by the direct effects on mitochondrial activity alone but must involve the inhibition of glycolysis as well. These inhibitory effects on energy production may contribute to the delayed toxicity of NO in vitro and in ischemic stroke. PMID- 9870947 TI - EAT-4, a homolog of a mammalian sodium-dependent inorganic phosphate cotransporter, is necessary for glutamatergic neurotransmission in caenorhabditis elegans. AB - The Caenorhabditis elegans gene eat-4 affects multiple glutamatergic neurotransmission pathways. We find that eat-4 encodes a protein similar in sequence to a mammalian brain-specific sodium-dependent inorganic phosphate cotransporter I (BNPI). Like BNPI in the rat CNS, eat-4 is expressed predominantly in a specific subset of neurons, including several proposed to be glutamatergic. Loss-of-function mutations in eat-4 cause defective glutamatergic chemical transmission but appear to have little effect on other functions of neurons. Our data suggest that phosphate ions imported into glutamatergic neurons through transporters such as EAT-4 and BNPI are required specifically for glutamatergic neurotransmission. PMID- 9870948 TI - Dendritic and postsynaptic protein synthetic machinery. AB - There is a growing body of evidence that local protein synthesis beneath synapses may provide a novel mechanism underlying plastic phenomena. In vivo and in vitro biochemical data show that dendrites can perform translation and glycosylation. Using antibodies directed against the eukaryotic protein synthetic machinery, we sought to identify the structures implicated in nonperinuclear translation, namely dendritic and postsynaptic protein synthesis. We performed a morphological and immunocytochemical analysis of ventromedial horn rat spinal cord neurons using both light and electron microscopy. We show at the cellular level that, in vivo, protein synthesis macrocomplexes (ribosomes and eIF-2) as well as the endomembranous system implicated in cotranslational and posttranslational modifications (endoplasmic reticulum and Golgi cisternae) penetrated some dendrites. Membrane-limited organelles of different shape and size are present close to the postsynaptic differentiations of most synapses, independently of their localization on the neuronal surface. We demonstrate (1) that some cisternae are immunoreactive for antibodies against ribosomal proteins and eIF-2, and (2) that markers of endoplasmic reticulum (BiP), intermediate compartment, and Golgi complex (rab1, CTR433, TGN38) label subsets of these subsynaptic organelles. Therefore, these findings indicate that synapses are equipped with the essential elements required for the synthesis and insertion of a well folded and glycosylated transmembrane protein. PMID- 9870949 TI - Regulation and immunohistochemical localization of betagamma-stimulated adenylyl cyclases in mouse hippocampus. AB - Specific forms of synaptic plasticity such as long-term potentiation (LTP) are modulated by or require increases in cAMP. The various adenylyl cyclase isoforms possess unique regulatory properties, and thus cAMP increases in a given cell type or tissue in response to converging signals are subject to the properties of the adenylyl cyclase isoforms expressed. In most tissues, adenylyl cyclase activity is stimulated by neurotransmitters or hormones via stimulatory G-protein (Gs)-coupled receptors and is inhibited via inhibitory G-protein (Gi)-linked receptors. However, in the hippocampus, stimulation of Gi-coupled receptors potentiates Gs-stimulated cAMP levels. This effect may be associated with the regulatory properties of adenylyl cyclase types 2 and 4 (AC2 and AC4), isoforms that are potentiated by the betagamma subunit of Gi in vitro. Although AC2 has been shown to be stimulated by betagamma in whole cells, reports describing the sensitivity of AC4 to betagamma in vivo have yet to emerge. Our results demonstrate that Gs-mediated stimulation of AC4 is potentiated by betagamma released from activated Gi-coupled receptors in intact human embryonic kidney (HEK) 293 cells. Furthermore, we show that the AC2 and AC4 proteins are expressed in the mouse hippocampal formation and that they colocalize with MAP2, a dendritic and/or postsynaptic marker. The presence of AC2 and AC4 in the hippocampus and the ability of each of these enzymes to detect coincident activation of Gs- and Gi-coupled receptors suggest that they may play a crucial role in certain forms of synaptic plasticity by coordinating such overlapping synaptic inputs. PMID- 9870950 TI - NMDA receptor-mediated control of presynaptic calcium and neurotransmitter release. AB - Before action potential-evoked Ca2+ transients, basal presynaptic Ca2+ concentration may profoundly affect the amplitude of subsequent neurotransmitter release. Reticulospinal axons of the lamprey spinal cord receive glutamatergic synaptic input. We have investigated the effect of this input on presynaptic Ca2+ concentrations and evoked release of neurotransmitter. Paired recordings were made between reticulospinal axons and the neurons that make axo-axonic synapses onto those axons. Both excitatory and inhibitory paired-cell responses were recorded in the axons. Excitatory synaptic inputs were blocked by the AMPA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 microM) and by the NMDA receptor antagonist 2-amino-5-phosphonopentanoate (AP-5; 50 microM). Application of NMDA evoked an increase in presynaptic Ca2+ in reticulospinal axons. Extracellular stimulation evoked Ca2+ transients in axons when applied either directly over the axon or lateral to the axons. Transients evoked by the two types of stimulation differed in magnitude and sensitivity to AP-5. Simultaneous microelectrode recordings from the axons during Ca2+ imaging revealed that stimulation of synaptic inputs directed to the axons evoked Ca2+ entry. By the use of paired-cell recordings between reticulospinal axons and their postsynaptic targets, NMDA receptor activation was shown to enhance evoked release of transmitter from the axons that received axoaxonic inputs. When the synaptic input to the axon was stimulated before eliciting an action potential in the axon, transmitter release from the axon was enhanced. We conclude that NMDA receptor-mediated input to reticulospinal axons increases basal Ca2+ within the axons and that this Ca2+ is sufficient to enhance release from the axons. PMID- 9870951 TI - Pituitary adenylate cyclase-activating polypeptide and melatonin in the suprachiasmatic nucleus: effects on the calcium signal transduction cascade. AB - The suprachiasmatic nucleus (SCN) harbors an endogenous oscillator generating circadian rhythms that are synchronized to the external light/dark cycle by photic information transmitted via the retinohypothalamic tract (RHT). The RHT has recently been shown to contain pituitary adenylate cyclase-activating polypeptide (PACAP) as neurotransmitter/neuromodulator. PACAPergic effects on cAMP-mediated signaling events in the SCN are restricted to distinct time windows and sensitive to melatonin. In neurons isolated from the SCN of neonatal rats we investigated by means of the fura-2 technique whether PACAP and melatonin also influence the intracellular calcium concentration ([Ca2+]i). PACAP elicited increases of [Ca2+]i in 27% of the analyzed neurons, many of which were also responsive to the RHT neurotransmitters glutamate and/or substance P. PACAP induced changes of [Ca2+]i were independent of cAMP, because they were not mimicked by forskolin or 8-bromo-cAMP. PACAP caused G-protein- and phospholipase C-mediated calcium release from inositol-trisphosphate-sensitive stores and subsequent protein kinase C-mediated calcium influx, demonstrated by treatment with GDP-beta-S, neomycin, U-73122, calcium-free saline, thapsigargin, bisindolylmaleimide, and chelerythrine. The calcium influx was insensitive to antagonists of voltage-gated calcium channels of the L-, N-, P-, Q- and T-type (diltiazem, nifedipine, verapamil, omega-conotoxin, omega-agatoxin, amiloride). Immunocytochemical characterization of the analyzed cells revealed that >50% of the PACAP-sensitive neurons were GABA-immunopositive. Our data demonstrate that in the SCN PACAP affects the [Ca2+]i, suggesting that different signaling pathways (calcium as well as cAMP) are involved in PACAPergic neurotransmission or neuromodulation. Melatonin did not interfere with calcium signaling, indicating that in SCN neurons the hormone primarily affects the cAMP signaling pathway. PMID- 9870952 TI - Prenatal development of retinogeniculate axons in the macaque monkey during segregation of binocular inputs. AB - In the fetal monkey the projections from the two eyes are initially completely intermingled within the dorsal lateral geniculate nucleus (DLGN) before separating into eye-specific layers (). To assess the cellular basis of this developmental process, we examined the morphological properties of individual retinogeniculate axons in prenatal monkeys of known gestational ages. The period studied spanned the time from when binocular overlap has been reported to be maximum, circa embryonic (E) day 77 through E112, when the segregation process is already largely completed in the caudal portion of the nucleus. Retinogeniculate fibers were labeled by making small deposits of DiI crystals into the fixed optic tract. After adequate time was allowed for diffusion of the tracer, fibers were visualized by confocal microscopy, and morphometric measures were made from photomontages. This revealed that retinogeniculate fibers in the embryonic monkey undergo continuous growth and elaboration during binocular overlap and subsequent segregation. Importantly, very few side-branches were found along the preterminal axon throughout the developmental period studied. Thus, restructuring of retinogeniculate fibers does not underlie the formation of eye-restricted projections in the primate. Rather, the results support the hypothesis that binocular segregation in the embryonic monkey is caused by the loss of retinal fibers that initially innervate inappropriate territories (). PMID- 9870953 TI - NMDA receptor-mediated refinement of a transient retinotectal projection during development requires nitric oxide. AB - A transient ipsilateral retinotectal projection is normally eliminated during embryonic development of the chick visual system. Administration of the NMDA receptor antagonist 5-methyl-10, 11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801) during the developmental period in which this projection normally disappears prevented its complete elimination. Previous studies showed that tectal cells express nitric oxide synthase during development, and blocking synthesis of nitric oxide also prevented elimination of the ipsilateral retinotectal projection. The effect of NMDA receptor blockade on nitric oxide synthase activity in tectal cells was assessed biochemically in chick embryos. Increasing concentrations of MK-801 resulted in a dose-dependent decrease in nitric oxide synthase activity. This result suggests that NMDA receptor activation can regulate nitric oxide synthase activity in the tectum. The degree of rescue of the ipsilateral retinotectal projection was compared in embryos treated either with MK-801 or with an inhibitor of nitric oxide synthesis, Nomega nitro-L-arginine (L-NoArg). At comparable levels of inhibition of nitric oxide synthesis, no significant difference was observed in the degree of rescue mediated by NMDA receptor blockade or nitric oxide synthesis blockade. These results suggest that NMDA receptor-mediated elimination of the ipsilateral retinotectal projection is completely mediated via nitric oxide. PMID- 9870954 TI - Stable properties of spontaneous EPSCs and miniature retinal EPSCs during the development of ON/OFF sublamination in the ferret lateral geniculate nucleus. AB - Retinal projections to the lateral geniculate nucleus (LGN) in ferrets progressively segregate into eye-specific laminae and subsequently into sublaminae that receive inputs from either ON-center or OFF-center afferents. To study the development of synaptic efficacy during a period of activity-dependent growth and reorganization in the CNS, we recorded spontaneous EPSCs (sEPSCs) from cells of the LGN during ON/OFF sublamination. We also examined retinal inputs specifically by stimulating the optic tract in the presence of strontium and recording evoked miniature EPSCs (emEPSCs). The rise times, areas, half-widths, and decay times of sEPSCs and emEPSCs and interevent intervals of sEPSCs recorded at the beginning of ON/OFF sublamination were not different from those recorded after its completion. Typically EPSC areas were small (10-20 fC) but varied greatly both within and between neurons. The frequency of sEPSCs was also quite variable, ranging from 0.2 to 5 Hz. sEPSCs were equivalent to miniature EPSCs recorded in the presence of tetrodotoxin, and both sEPSCs and emEPSCs were CNQX sensitive. No difference was observed between sEPSCs recorded at room temperature and those recorded at 34 degreesC, and strontium could be substituted for calcium with no effect on sEPSC shape. These data argue for a remarkable stability in the components of at least AMPA-mediated synaptic transmission during a period of major synaptic rearrangement in the LGN. PMID- 9870955 TI - Neuronal degeneration in canine narcolepsy. AB - Narcolepsy is a lifelong illness characterized by persistent sleepiness, hypnagogic hallucinations, and episodes of motor paralysis called cataplexy. We have tested the hypothesis that a transient neurodegenerative process is linked to symptom onset. Using the amino-cupric silver stain on brain sections from canine narcoleptics, we found elevated levels of axonal degeneration in the amygdala, basal forebrain (including the nucleus of the diagonal band, substantia innominata, and preoptic region), entopeduncular nucleus, and medial septal region. Reactive neuronal somata, an indicator of neuronal pathology, were found in the ventral amygdala. Axonal degeneration was maximal at 2-4 months of age. The number of reactive cells was maximal at 1 month of age. These degenerative changes precede or coincide with symptom onset. The forebrain degeneration that we have observed can explain the major symptoms of narcolepsy. PMID- 9870956 TI - Absence of the p75 neurotrophin receptor alters the pattern of sympathosensory sprouting in the trigeminal ganglia of mice overexpressing nerve growth factor. AB - Sympathetic axons invade the trigeminal ganglia of mice overexpressing nerve growth factor (NGF) (NGF/p75(+/+) mice) and surround sensory neurons having intense NGF immunolabeling; the growth of these axons appears to be directional and specific (). In this investigation, we provide new insight into the neurochemical features and receptor requirements of this sympathosensory sprouting. Using double-antigen immunohistochemistry, we demonstrate that virtually all (98%) trigeminal neurons that exhibit a sympathetic plexus are trk tyrosine kinase receptor (trkA)-positive. In addition, the majority (86%) of those neurons enveloped by sympathetic fibers is also calcitonin gene-related peptide (CGRP)-positive; a smaller number of plexuses (14%) surrounded other somata lacking this neuropeptide. Our results show that sympathosensory interactions form primarily between noradrenergic sympathetic efferents and the trkA/CGRP-expressing sensory somata. To assess the contribution of the p75 neurotrophin receptor (p75(NTR)) in sympathosensory sprouting, a hybrid strain of mice was used that overexpresses NGF but lacks p75(NTR) expression (NGF/p75(-/-) mice). The trigeminal ganglia of NGF/p75(-/-) mice, like those of NGF/p75(+/+) mice, have increased levels of NGF protein and display a concomitant ingrowth of sympathetic axons. In contrast to the precise pattern of sprouting seen in the ganglia of NGF/p75(+/+) mice, sympathetic axons course randomly throughout the ganglionic neuropil of NGF/p75(-/-) mice, forming few perineuronal plexuses. Our results indicate that p75(NTR) is not required to initiate or sustain the growth of sympathetic axons into the NGF-rich trigeminal ganglia but rather plays a role in regulating the directional patterns of axon growth. PMID- 9870957 TI - Oscillatory coupling of hippocampal pyramidal cells and interneurons in the behaving Rat. AB - We examined whether excitation and inhibition are balanced in hippocampal cortical networks. Extracellular field and single-unit activity were recorded by multiple tetrodes and multisite silicon probes to reveal the timing of the activity of hippocampal CA1 pyramidal cells and classes of interneurons during theta waves and sharp wave burst (SPW)-associated field ripples. The somatic and dendritic inhibition of pyramidal cells was deduced from the activity of interneurons in the pyramidal layer [int(p)] and in the alveus and st. oriens [int(a/o)], respectively. Int(p) and int(a/o) discharged an average of 60 and 20 degrees before the population discharge of pyramidal cells during the theta cycle, respectively. SPW ripples were associated with a 2.5-fold net increase of excitation. The discharge frequency of int(a/o) increased, decreased ("anti-SPW" cells), or did not change ("SPW-independent" cells) during SPW, suggesting that not all interneurons are innervated by pyramidal cells. Int(p) either fired together with (unimodal cells) or both before and after (bimodal cells) the pyramidal cell burst. During fast-ripple oscillation, the activity of interneurons in both the int(p) and int(a/o) groups lagged the maximum discharge probability of pyramidal neurons by 1-2 msec. Network state changes, as reflected by field activity, covaried with changes in the spike train dynamics of single cells and their interactions. Summed activity of parallel-recorded interneurons, but not of pyramidal cells, reliably predicted theta cycles, whereas the reverse was true for the ripple cycles of SPWs. We suggest that network-driven excitability changes provide temporal windows of opportunity for single pyramidal cells to suppress, enable, or facilitate selective synaptic inputs. PMID- 9870958 TI - The mammalian staufen protein localizes to the somatodendritic domain of cultured hippocampal neurons: implications for its involvement in mRNA transport. AB - In hippocampal neurons, certain mRNAs have been found in dendrites (), and their localization and translation have been implicated in synaptic plasticity (). One attractive candidate to achieve transport of mRNAs into dendrites is Staufen (Stau), a double-stranded RNA-binding protein, which plays a pivotal role in mRNA transport, localization, and translation in Drosophila (). Using antibodies raised against a peptide located in the RNA-binding domain IIa and a polyclonal antibody raised against a recently cloned human Staufen homolog, we identify a 65 kDa rat homolog in cultured rat hippocampal neurons. In agreement with the exclusive somatodendritic localization of mRNAs in these cells, we find that Staufen is restricted to the same domain. By immunoelectron microscopy, we show enrichment of the mammalian homolog of Stau (mStau) in the vicinity of smooth endoplasmic reticulum and microtubules near synaptic contacts. Finally, the association of the mStau with neuronal mRNAs is suggested by the colocalization with ribonucleoprotein particles specifically in distal dendrites known to contain mRNA, ribosomes, and translation factors (). These results suggest a role for mStau in the polarized transport and localization of mRNAs in mammalian neurons. PMID- 9870959 TI - Peripheral synapses at identified mechanosensory neurons in spiders: three dimensional reconstruction and GABA immunocytochemistry. AB - The mechanosensory organs of arachnids receive diverse peripheral inputs. Little is known about the origin, distribution, and function of these chemical synapses, which we examined in lyriform slit sense organ VS-3 of the spider Cupiennius salei. The cuticular slits of this organ are each associated with two large bipolar mechanosensory neurons with different adaptation rates. With intracellular recording, we have now been able to correlate directly the staining intensity of a neuron for acetylcholinesterase with its adaptation rate, thus allowing us simply to stain a neuron to identify its functional type. All rapidly adapting neurons stain more heavily than slowly adapting neurons. Immunostaining of whole-mount preparations reveals GABA-like immunoreactive fibers forming numerous varicosities at the surface of all sensory neurons in VS-3; peripheral GABA-like immunoreactive somata are lacking. Sectioning the leg nerve procures rapid degeneration of most fiber profiles, confirming that the fibers are efferent. Punctate synapsin-like immunoreactivity colocalizes to these varicosities, although some synapsin-like immunoreactive puncta are GABA immunonegative. Fibers with similar immunoreactivities are also associated with trichobothria, tactile hairs, internal joint receptors, i.e. other types of spider mechanosensory organs. In organ VS-3, immunoreactivity is most dense across the initial axon segment. The exact distribution of peripheral synapses was reconstructed from a 10-microm-long electron micrograph series of the dendritic, somatic, and initial axon regions of acetylcholinesterase-stained VS-3 neurons. These reveal a pattern similar to that of the synapsin-like immunoreactivity. Two different types of synapse were distinguished on the basis of their presynaptic vesicle populations. Many peripheral synapses thus appear to derive from efferent GABA-like immunoreactive fibers and probably provide centrifugal inhibitory control of primary mechanosensory activities. PMID- 9870960 TI - Homing in pigeons: the role of the hippocampal formation in the representation of landmarks used for navigation. AB - When given repeated training from a location, homing pigeons acquire the ability to use familiar landmarks to navigate home. Both control and hippocampal-lesioned pigeons succeed in learning to use familiar landmarks for homing. However, the landmark representations that guide navigation are strikingly different. Control and hippocampal-lesioned pigeons were initially given repeated training flights from two locations. On subsequent test days from the two training locations, all pigeons were rendered anosmic to eliminate use of their navigational map and were phase- or clock-shifted to examine the extent to which their learned landmark representations were dependent on the use of the sun as a compass. We show that control pigeons acquire a landmark representation that allows them to directly use landmarks without reference to the sun to guide their flight home, called "pilotage". Hippocampal-lesioned birds only learn to use familiar landmarks at the training location to recall the compass direction home, based on the sun, flown during training, called "site-specific compass orientation." The results demonstrate that for navigation of 20 km or more in a natural field setting, the hippocampal formation is necessary if homing pigeons are to learn a spatial representation based on numerous independent landmark elements that can be used to directly guide their return home. PMID- 9870961 TI - Computation of inertial motion: neural strategies to resolve ambiguous otolith information. AB - According to Einstein's equivalence principle, inertial accelerations during translational motion are physically indistinguishable from gravitational accelerations experienced during tilting movements. Nevertheless, despite ambiguous sensory representation of motion in primary otolith afferents, primate oculomotor responses are appropriately compensatory for the correct translational component of the head movement. The neural computational strategies used by the brain to discriminate the two and to reliably detect translational motion were investigated in the primate vestibulo-ocular system. The experimental protocols consisted of either lateral translations, roll tilts, or combined translation tilt paradigms. Results using both steady-state sinusoidal and transient motion profiles in darkness or near target viewing demonstrated that semicircular canal signals are necessary sensory cues for the discrimination between different sources of linear acceleration. When the semicircular canals were inactivated, horizontal eye movements (appropriate for translational motion) could no longer be correlated with head translation. Instead, translational eye movements totally reflected the erroneous primary otolith afferent signals and were correlated with the resultant acceleration, regardless of whether it resulted from translation or tilt. Therefore, at least for frequencies in which the vestibulo-ocular reflex is important for gaze stabilization (>0.1 Hz), the oculomotor system discriminates between head translation and tilt primarily by sensory integration mechanisms rather than frequency segregation of otolith afferent information. Nonlinear neural computational schemes are proposed in which not only linear acceleration information from the otolith receptors but also angular velocity signals from the semicircular canals are simultaneously used by the brain to correctly estimate the source of linear acceleration and to elicit appropriate oculomotor responses. PMID- 9870962 TI - A nonphotic stimulus inverts the diurnal-nocturnal phase preference in Octodon degus. AB - Mechanisms differentiating diurnal from nocturnal species are thought to be innate components of the circadian timekeeping system and may be located downstream from the circadian pacemaker within the suprachiasmatic nucleus (SCN) of the hypothalamus. In the present study, we found that the dominant phase of behavioral activity and body temperature (Tb) is susceptible to modification by a specific modality of behavioral activity (wheel-running activity) in Octodon degus, a mammal that exhibits multiple chronotypes. Seven Octodon degus exhibited diurnal Tb and locomotor activity (LMA) circadian rhythms while entrained to a 24 h light/dark cycle (LD 12:12). When the diurnal animals were provided unrestricted access to a running wheel, the overt daily rhythms in these animals inverted to nocturnal. This nocturnal pattern was sustained in constant darkness and returned to diurnal after removal of the running wheel. Six additional animals exhibited nocturnal chronotypes in LD 12:12 regardless of access to running wheels. Wheel-running activity inverted the phase preference in the diurnal animals without changing the 24 hr mean LMA or Tb levels. Because wheel running did not increase the amplitude of the pre-existing diurnal pattern, simple masking effects on LMA and Tb cannot explain the rhythm inversion. The diurnal-nocturnal inversion occurred without reversing crepuscular-timed episodes of activity, suggesting that diurnal or nocturnal phase preference is controlled separately from the intrinsic timing mechanisms within the SCN and can be dependent on behavioral or environmental factors. PMID- 9870963 TI - Developmental dissociation of serotonin-induced spike broadening and synaptic facilitation in Aplysia sensory neurons. AB - In sensory neurons (SNs) of adult Aplysia, serotonin (5-HT)-induced spike broadening has long been implicated as important for synaptic facilitation [spike duration-dependent (SDD) facilitation], particularly at nondepressed synapses. At depressed synapses, spike broadening has less impact on synaptic facilitation; under these conditions, 5-HT induces a spike duration-independent (SDI) form of facilitation (). It has been difficult to dissociate clearly the cellular mechanisms underlying these two forms of facilitation. However, the observation that a major form of spike broadening emerges late in juvenile development () provides a unique opportunity to examine the relationship between spike broadening and synaptic facilitation in juvenile Aplysia. We have identified three forms of synaptic plasticity in juvenile Aplysia: homosynaptic depression, SDD facilitation, and SDI facilitation. We show that homosynaptic depression is fully developed in the juvenile and that 5-HT reliably induces synaptic facilitation at depressed synapses. However, in nondepressed synapses, 5-HT induced facilitation is not reliable. Further analysis revealed that the relationship between spike broadening and synaptic facilitation for nondepressed synapses is the inverse of that in adults. Surprisingly, in juveniles, minor spike broadening induced by 5-HT results in significant synaptic facilitation, whereas major spike broadening, when it occurs, does not. These results suggest a model in which juvenile synapses predominantly use SDI facilitation, and with the emergence of major spike broadening, a developmentally transient inhibitory process emerges. This inhibitory process seems to be independent of major spike broadening induced by 5-HT because directly broadening the spike with 4 aminopyridine induces adult-like SDD synaptic facilitation. Finally, in the adult, the inhibitory process is either lost or masked, and SDD facilitation predominates at nondepressed synapses. PMID- 9870964 TI - Cellular traces of behavioral classical conditioning can be recorded at several specific sites in a simple nervous system. AB - We used a behavioral learning paradigm followed by electrophysiological analysis to find sites in the Lymnaea feeding network in which electrical changes could be recorded after appetitive conditioning. Specifically, we analyzed conditioning induced changes in cellular responses in the mechanosensory conditioned stimulus (CS) pathway, in the central pattern generator (CPG) network, and in feeding motoneurons. During training, experimental animals received 15 pairings of lip touch (the CS) with sucrose (the unconditioned stimulus, US). Control animals received 15 random CS and US presentations. Electrophysiological tests on semi intact preparations made from conditioned animals demonstrated a network correlate of the overall feeding conditioned response, a touch-evoked CPG-driven fictive feeding rhythm. At the motoneuronal level, we found significant conditioning-induced increases in the amplitude of an early touch-evoked EPSP and spike activity, recorded from the B3 feeding motoneuron. Increases in EPSP amplitude and motoneuronal spike activity could occur independently of conditioned fictive feeding. These changes in response recorded at the level of CPG interneurons, and motoneurons were preceded by changes recorded in the CS pathway. This was demonstrated by recording a conditioning-induced increase in the number of touch-evoked spikes in the cerebrobuccal connective, which forms part of the CS pathway. The finding that electrophysiological changes after conditioning can be recorded at multiple sites in this simple system provided an important intermediate level of analysis between whole animal behavior and cellular studies on the synaptic sites of plasticity. PMID- 9870965 TI - High-frequency auditory feedback is not required for adult song maintenance in Bengalese finches. AB - Male Bengalese finches do not normally change their vocal patterns in adulthood; song is stereotyped and stable over time. Adult song maintenance requires auditory feedback. If adults are deafened, song will degrade within 1 week. We tested whether feedback of all sound frequencies is required for song maintenance. The avian basilar papilla is tonotopically organized; hair cells in the basal region encode high frequencies, and low frequencies are encoded in progressively apical regions. We restricted the spectral range of feedback available to a bird by killing either auditory hair cells encoding higher frequencies or those encoding both high and low frequencies and documented resultant changes in song. Birds were treated with either Amikacin alone to kill high-frequency hair cells or Amikacin and sound exposure to target hair cells across the entire papilla. During treatment, song was recorded from all birds weekly. After treatment and song recording, evoked-potential audiograms were evaluated on each bird, and papillas were evaluated by scanning electron microscopy. Results showed that hair cell damage over 46-63% of the basal papilla and the corresponding high-frequency hearing loss had no effect on song structure. In birds with hair cell damage extending further into the apical region of the papilla and corresponding low-frequency and high-frequency hearing loss, song degradation occurred within 1 week of beginning treatment and was comparable with degradation after surgical deafening. We conclude that either low frequency spectral cues or temporal cues via feedback of the song amplitude envelope are sufficient for song maintenance in adult Bengalese finches. PMID- 9870966 TI - The role of the intergeniculate leaflet in entrainment of circadian rhythms to a skeleton photoperiod. AB - Mammalian circadian rhythms are synchronized to environmental light/dark (LD) cycles via daily phase resetting of the circadian clock in the suprachiasmatic nucleus (SCN). Photic information is transmitted to the SCN directly from the retina via the retinohypothalamic tract (RHT) and indirectly from the retinorecipient intergeniculate leaflet (IGL) via the geniculohypothalamic tract (GHT). The RHT is thought to be both necessary and sufficient for photic entrainment to standard laboratory light/dark cycles. An obligatory role for the IGL-GHT in photic entrainment has not been demonstrated. Here we show that the IGL is necessary for entrainment of circadian rhythms to a skeleton photoperiod (SPP), an ecologically relevant lighting schedule congruous with light sampling behavior in nocturnal rodents. Rats with bilateral electrolytic IGL lesions entrained normally to lighting cycles consisting of 12 hr of light followed by 12 hr of darkness, but exhibited free-running rhythms when housed under an SPP consisting of two 1 hr light pulses given at times corresponding to dusk and dawn. Despite IGL lesions and other damage to the visual system, the SCN displayed normal sensitivity to the entraining light, as assessed by light induced Fos immunoreactivity. In addition, all IGL-lesioned, free-running rats showed masking of the body temperature rhythm during the SPP light pulses. These results show that the integrity of the IGL is necessary for entrainment of circadian rhythms to a lighting schedule like that experienced by nocturnal rodents in the natural environment. PMID- 9870967 TI - Relationship between afferent and central temporal patterns in the locust olfactory system. AB - Odors evoke synchronized oscillations and slow temporal patterns in antennal lobe neurons and fast oscillations in the mushroom body local field potential (LFP) of the locust. What is the contribution of primary afferents in the generation of these dynamics? We addressed this question in two ways. First, we recorded odor evoked afferent activity in both isolated antennae and intact preparations. Odor evoked population activity in the antenna and the antennal nerve consisted of a slow potential deflection, similar for many odors. This deflection contained neither oscillatory nor odor-specific slow temporal patterns, whereas simultaneously recorded mushroom body LFPs exhibited clear 20-30 Hz oscillations. This suggests that the temporal patterning of antennal lobe and mushroom body neurons is generated downstream of the olfactory receptor axons. Second, we electrically stimulated arrays of primary afferents in vivo. A brief shock to the antennal nerve produced compound PSPs in antennal lobe projection neurons, with two peaks at an approximately 50 msec interval. Prolonged afferent stimulation with step, ramp, or slow sine-shaped voltage waveforms evoked sustained 20-30 Hz oscillations in projection neuron membrane potential and in the mushroom body LFP. Projection neuron and mushroom body oscillations were phase-locked and reliable across trials. Synchronization of projection neurons was seen directly in paired intracellular recordings. Pressure injection of picrotoxin into the antennal lobe eliminated the oscillations evoked by electrical stimulation. Different projection neurons could express different temporal patterns in response to the same electrical stimulus, as seen for odor-evoked responses. Conversely, individual projection neurons could express different temporal patterns of activity in response to step stimulation of different spatial arrays of olfactory afferents. These patterns were reliable and remained distinct across different stimulus intensities. We conclude that oscillatory synchronization of olfactory neurons originates in the antennal lobe and that slow temporal patterns in projection neurons can arise in the absence of temporal patterning of the afferent input. PMID- 9870968 TI - The modulation of presynaptic inhibition in single muscle primary afferents during fictive locomotion in the cat. AB - The aim of this study is to understand the functional organization of presynaptic inhibition in muscle primary afferents during locomotion. Primary afferent depolarization (PAD) associated with presynaptic inhibition was recorded intra axonally in identified afferents from various hindlimb muscles in L6-L7 spinal segments during fictive locomotion in the decerebrate cat. PADs were evoked by the stimulation of peripheral muscle nerves and were averaged in the different epochs of the fictive step cycle. Fifty-three trials recorded from 39 muscle axons (37 from group I and two from group II) were retained for analysis. The results showed that there was a significant phase-dependent modulation of PAD amplitude (p < 0.05) in a majority of muscle afferents (30 of 39, 77%). However, not all stimulated nerves led to significantly modulated PADs in a given axon (36 of 53 trials, 68%). We also observed that the pattern of modulation (phase for maximum and minimum PAD amplitude and the depth of modulation) varied with each recorded afferent, as well as with each stimulated nerve. We further evaluated the effect of PAD modulation on the phasic transmission of the monosynaptic reflex (MSR) and found that PADs decreased the MSR amplitude in all phases of the fictive step cycle, independent of the PAD pattern in individual group I fibers. We conclude that (1) PAD modulation patterns of all group I fibers contacting motoneurons led to an overall reduction in monosynaptic transmission, and (2) individual PAD patterns could participate in the control of transmission in specific reflex pathways during locomotion. PMID- 9870969 TI - Velocity invariance of receptive field structure in somatosensory cortical area 3b of the alert monkey. AB - This is the second in a series of studies of the neural representation of tactile spatial form in cortical area 3b of the alert monkey. We previously studied the spatial structure of 330 area 3b neuronal receptive fields (RFs) on the fingerpad with random dot patterns scanned at one velocity (40 mm/sec; ). Here, we analyze the temporal structure of 84 neuronal RFs by studying their spatial structure at three scanning velocities (20, 40, and 80 mm/sec). As in the previous study, most RFs contained a single, central, excitatory region and one or more surrounding or flanking inhibitory regions. The mean time delay between skin stimulation and its excitatory effect was 15.5 msec. Except for differences in mean rate, each neuron's response and the spatial structure of its RF were essentially unaffected by scanning velocity. This is the expected outcome when excitatory and inhibitory effects are brief and synchronous. However, that interpretation is consistent neither with the reported timing of excitation and inhibition in somatosensory cortex nor with the third study in this series, which investigates the effect of scanning direction and shows that one component of inhibition lags behind excitation. We reconcile these observations by showing that overlapping (in field) inhibition delayed relative to excitation can produce RF spatial structure that is unaffected by changes in scanning velocity. Regardless of the mechanisms, the velocity invariance of area 3b RF structure is consistent with the velocity invariance of tactile spatial perception (e.g., roughness estimation and form recognition). PMID- 9870970 TI - Pain pathways involved in fear conditioning measured with fear-potentiated startle: lesion studies. AB - It is well established that the basolateral amygdala is critically involved in the association between an unconditioned stimulus (US), such as a foot shock, and a conditioned stimulus (CS), such as a light, during classic fear conditioning. However, little is known about how the US (pain) inputs are relayed to the basolateral amygdala. The present studies were designed to define potential US pathways to the amygdala using lesion methods. Electrolytic lesions before or after training were placed in caudal granular/dysgranular insular cortex (IC) alone or in conjunction with the posterior intralaminar nuclei of the thalamus (PoT/PIL), and the effects on fear conditioning were examined. Pretraining lesions of both IC and PoT/PIL, but not lesions of IC alone, blocked the acquisition of fear-potentiated startle. However, post-training combined lesions of IC and PoT/PIL did not prevent expression of conditioned fear. Given that previous studies have shown that lesions of PoT/PIL alone had no effect on acquisition of conditioned fear, these results suggest that two parallel cortical (insula-amygdala) and subcortical (PoT/PIL-amygdala) pathways are involved in relaying shock information to the basolateral amygdala during fear conditioning. PMID- 9870971 TI - Effects of attention on orientation-tuning functions of single neurons in macaque cortical area V4. AB - We examined how attention affected the orientation tuning of 262 isolated neurons in extrastriate area V4 and 135 neurons in area V1 of two rhesus monkeys. The animals were trained to perform a delayed match-to-sample task in which oriented stimuli were presented in the receptive field of the neuron being recorded. On some trials the animals were instructed to pay attention to those stimuli, and on other trials they were instructed to pay attention to other stimuli outside the receptive field. In this way, orientation-tuning curves could be constructed from neuronal responses collected in two behavioral states: one in which those stimuli were attended by the animal and one in which those stimuli were ignored by the animal. We fit Gaussians to the neuronal responses to twelve different orientations for each behavioral state. Although attention enhanced the responses of V4 neurons (median 26% increase) and V1 neurons (median 8% increase), selectivity, as measured by the width of its orientation-tuning curve, was not systematically altered by attention. The effects of attention were consistent with a multiplicative scaling of the driven response to all orientations. We also found that attention did not cause systematic changes in the undriven activity of the neurons. PMID- 9870972 TI - UV- and midwave-sensitive cone-driven retinal responses of the mouse: a possible phenotype for coexpression of cone photopigments. AB - Molecular biological, histological and flicker electroretinographic results have established that mice have two cone photopigments, one peaking near 350 nm (UV cone pigment) and a second near 510 nm [midwave (M)-cone pigment]. The goal of this investigation was to measure the action spectra and absolute sensitivities of the UV-cone- and M-cone-driven b-wave responses of C57BL/6 mice. To achieve this goal, we suppressed rod-driven signals with steady or flashed backgrounds and obtained intensity-response relations for cone-driven b-waves elicited by narrowband flashes between 340 and 600 nm. The derived cone action spectra can be described as retinal1 pigments with peaks at 355 and 508 nm. The UV peak had an absolute sensitivity of approximately 8 nV/(photon microm2) at the cornea, approximately fourfold higher than the M peak. In an attempt to isolate UV-cone driven responses, it was discovered that an orange conditioning flash (lambda > 530 nm) completely suppressed ERG signals driven by both M pigment- and UV pigment-containing cones. Analysis showed that the orange flash could not have produced a detectable response in the UV-cone pathway were their no linkage between M pigment- and UV pigment-generated signals. Because cones containing predominantly the UV and M pigments have been shown to be located largely in separate parts of the mouse retina (), the most probable linkage is coexpression of M pigment in cones primarily expressing UV pigment. New histological evidence supports this interpretation (). Our data are consistent with an upper bound of approximately 3% coexpression of M pigment in the cones that express mostly the UV pigment. PMID- 9870973 TI - Facilitation of sexual behavior and enhanced dopamine efflux in the nucleus accumbens of male rats after D-amphetamine-induced behavioral sensitization. AB - Behavioral sensitization caused by repeated and intermittent administration of psychostimulants, such as cocaine and D-amphetamine, is accompanied by enhanced function in limbic-motor circuitry that is involved in the generation of motivated behavior. The present microdialysis study investigated the effect of D amphetamine-induced sensitization on dopamine (DA) efflux in the nucleus accumbens (NAC) of male rats during sexual behavior. Male rats were given one injection of D-amphetamine (1.5 mg/kg, i.p.) or saline every other day for a total of 10 injections. Three weeks after discontinuation of drug treatment, rats were tested for sexual behavior during a test in which microdialysis was performed. There was an augmented efflux of DA in the NAC of D-amphetamine sensitized rats compared with nonsensitized control rats when a receptive female was present behind a screen (35 vs 17%). Sensitized rats exhibited facilitated sexual behavior when the screen was removed, as indicated by a significantly shorter latency to mount and an overall increase in the amount of copulatory behavior. Although there was a significant increase in NAC DA concentrations from baseline in both sensitized and nonsensitized rats during copulation, there was a greater increase in DA efflux in the NAC of sensitized rats during the first 10 min copulatory sample (60 vs 37%). These results demonstrate that behavioral sensitization caused by repeated psychostimulant administration can "cross sensitize" to a natural behavior, such as sex, and that increased NAC DA release may contribute to the facilitation of appetitive and consummatory aspects of this behavior. PMID- 9870974 TI - 2,3-Dihydroxy-6-nitro-7-sulfamoyl-benzo(f)quinoxaline reduces glial loss and acute white matter pathology after experimental spinal cord contusion. AB - Focal microinjection of 2, 3-dihyro-6-nitro-7-sulfamoyl-benzo(f)quinoxaline (NBQX), an antagonist of the AMPA/kainate subclass of glutamate receptors, reduces neurological deficits and tissue loss after spinal cord injury. Dose dependent sparing of white matter is seen at 1 month after injury that is correlated to the dose-related reduction in chronic functional deficits. To determine whether NBQX exerts an acute effect on white matter pathology, female, adult Spague Dawley rats were subjected to a standardized weight drop contusion at T-8 (10 gm x 2.5 cm) and NBQX (15 nmol) or vehicle (VEH) solution focally injected into the injury site 15 min later. At 4 and 24 hr, tissue from the injury epicenter was processed for light and electron microscopy, and the histopathology of ventromedial white matter was compared. The axonal injury index, a quantitative representation of axoplasmic and myelinic pathologies, was significantly lower in the NBQX group at 4 hr (2.7 +/- 0.24, mean +/- SE) and 24 hr (1.4 +/- 0.19) than in VEH controls (3.8 +/- 0.33 and 2.1 +/- 0.20, respectively). Counts of glial cell nuclei indicated a loss of at least 60% at 4 and 24 hr after injury in the VEH group compared with uninjured controls. NBQX treatment reduced this glial loss by half. Immunohistochemistry revealed that the spared glia were primarily oligodendrocytes. Thus, the chronic effects of NBQX in reducing white matter loss after spinal cord injury appear to be attributable to the reduction of acute pathology and may be mediated through the protection of glia, particularly oligodendrocytes. PMID- 9870975 TI - Contributions of social cues and photoperiod to seasonal plasticity in the adult avian song control system. AB - In seasonally breeding birds, the vernal growth of the song system is thought to result primarily from increased daylength and the associated increase in circulating testosterone. Other environmental factors such as social cues between mates influence the timing of reproduction, but less is known about how social cues might affect the song system and song behavior. We used white-crowned sparrows (Zonotrichia leucophrys gambelii) to test the hypothesis that the presence of a female in breeding condition influences song nuclei and song behavior of adult males. There were four treatment groups: (1) eight males housed individually in the same room on long days and paired with estradiol-implanted females; (2) eight males housed similarly on long days but without females; (3) four males isolated on long days; and (4) four males isolated on short days. The volumes of two song nuclei, HVc and RA, were significantly larger in males housed with females than in any other treatment group. Males isolated on short days had smaller HVc, RA, and area X volumes than all other groups. The volumes of Rt (a thalamic nucleus not involved in song) and the telencephalon did not differ among groups. Plasma androgen levels did not differ among the three long-day, social treatment groups at the times sampled, but were lower in the short-day isolates. Males paired with females sang at a higher maximum rate than males housed together, who sang at a higher rate than long-day isolates. These results suggest that seasonal plasticity in the adult song system is influenced by social cues. PMID- 9870976 TI - Neuropharmacological dissection of placebo analgesia: expectation-activated opioid systems versus conditioning-activated specific subsystems. AB - We investigated the mechanisms underlying the activation of endogenous opioids in placebo analgesia by using the model of human experimental ischemic arm pain. Different types of placebo analgesic responses were evoked by means of cognitive expectation cues, drug conditioning, or a combination of both. Drug conditioning was performed by means of either the opioid agonist morphine hydrochloride or the nonopioid ketorolac tromethamine. Expectation cues produced placebo responses that were completely blocked by the opioid antagonist naloxone. Expectation cues together with morphine conditioning produced placebo responses that were completely antagonized by naloxone. Morphine conditioning alone (without expectation cues) induced a naloxone-reversible placebo effect. By contrast, ketorolac conditioning together with expectation cues elicited a placebo effect that was blocked by naloxone only partially. Ketorolac conditioning alone produced placebo responses that were naloxone-insensitive. Therefore, we evoked different types of placebo responses that were either naloxone-reversible or partially naloxone-reversible or, otherwise, naloxone-insensitive, depending on the procedure used to evoke the placebo response. These findings show that cognitive factors and conditioning are balanced in different ways in placebo analgesia, and this balance is crucial for the activation of opioid or nonopioid systems. Expectation triggers endogenous opioids, whereas conditioning activates specific subsystems. In fact, if conditioning is performed with opioids, placebo analgesia is mediated via opioid receptors, if conditioning is performed with nonopioid drugs, other nonopioid mechanisms result to be involved. PMID- 9870977 TI - Functionally dissociating aspects of event memory: the effects of combined perirhinal and postrhinal cortex lesions on object and place memory in the rat. AB - Reciprocal interactions between the hippocampus and the perirhinal and parahippocampal cortices form core components of a proposed temporal lobe memory system. For this reason, the involvement of the hippocampus in event memory is thought to depend on its connections with these cortical areas. Contrary to these predictions, we found that NMDA-induced lesions of the putative rat homologs of these cortical areas (perirhinal plus postrhinal cortices) did not impair performance on two allocentric spatial tasks highly sensitive to hippocampal dysfunction. Remarkably, for one of the tasks there was evidence of a facilitation of performance. The same cortical lesions did, however, disrupt spontaneous object recognition and object discrimination reversal learning but spared initial acquisition of the discrimination. This pattern of results reveals important dissociations between different aspects of memory within the temporal lobe. Furthermore, it shows that the perirhinal-postrhinal cortex is not a necessary route for spatial information reaching the hippocampus and that object familiarity-novelty detection depends on different neural substrates than do other aspects of event memory. PMID- 9870979 TI - Passing the torch PMID- 9870980 TI - Responsible conduct in publishing PMID- 9870981 TI - Responsible conduct regarding scientific communication PMID- 9870978 TI - Identification of the receptor subtype involved in the analgesic effect of neurotensin. AB - The neuropeptide neurotensin (NT) elicits hypothermic and naloxone-insensitive analgesic responses after brain injection. Recent pharmacological evidence obtained with NT agonists and antagonists suggests that these effects are mediated by a receptor distinct from the initially cloned high-affinity NT receptor (NTR1). The recent cloning of a second NT receptor (NTR2) prompted us to evaluate its role in NT-induced analgesia. Intracerebroventricular injections in mice of two different antisense oligodeoxynucleotides from the NTR2 markedly decreased NTR2 mRNA and protein and reduced NT-induced analgesia. This effect was specific, because NTR1 levels were unaffected, and sense or scramble oligodeoxynucleotides had no effect. Structure-activity studies revealed a close correlation between the analgesic potency of NT analogs and their affinity for the NTR2 and disclosed potent and selective agonists of this receptor. These data confirm that NTR1 is involved in the NT-elicited turning behavior and demonstrate that the NTR2 mediates NT-induced analgesia. PMID- 9870982 TI - Pentachlorophenol and pentachloroanisole in oil samples associated with the toxic oil syndrome. PMID- 9870983 TI - Method for the determination of PCB congeners and chlorinated pesticides in human blood serum. PMID- 9870984 TI - Cytotoxicity of lindane and paraquat to human hepatoma cell lines. PMID- 9870985 TI - Interlaboratory comparison of endotoxin analyses in occupational exposure. PMID- 9870986 TI - Lead and cadmium in Ethiopian vegetables. PMID- 9870987 TI - Degradation of sodium monofluoroacetate (1080) and fluorocitrate in water. PMID- 9870988 TI - Metals in surficial sediments of the north-west Irish Sea. PMID- 9870989 TI - Bacterial degradation of chlorpyrifos in pure cultures and in soil. PMID- 9870990 TI - Agricultural inputs of pesticide residues to stream and pond sediments in a small catchment in southern Sweden. PMID- 9870992 TI - Nitrate, ammonium, and phosphate uptake by the immobilized cells of Dunaliella salina. PMID- 9870991 TI - Effects of zinc exposure on oxygen consumption and gill Na+, K+-ATPase of the estuarine crab Chasmagnathus granulata Dana, 1851 (Decapoda-grapsidae). PMID- 9870993 TI - Toxicity assays and bioconcentration of mercury in bacteria selected from marine environments. PMID- 9870994 TI - Acclimation of algal species following exposure to phenol. PMID- 9870995 TI - Human placental microsomal activation and DNA adduction by air pollutants. PMID- 9870996 TI - Methyl bromide and bromide ion in soil after experimental greenhouse fumigation. PMID- 9870997 TI - Degradation of explosive 2,4,6-trinitrotoluene by s-triazine degrading bacterium isolated from contaminated soil. PMID- 9870998 TI - Persistence of phorate and carbofuran in relation to their effect on the mineralization of C, N, and P in alluvial soil. PMID- 9870999 TI - Pesticide residues in marketed sesame. PMID- 9871000 TI - Naturally occurring radium-226 and radium-228 in water supplies of Michigan. PMID- 9871001 TI - Lead and cadmium in red deer and wild boar from Sierra Morena Mountains (Andalusia, Spain). PMID- 9871002 TI - Ecotoxicity of pulp mill effluents from different prebleaching processes. PMID- 9871004 TI - Ecotoxicity of river and spring sediment along the Hanford Reach. PMID- 9871003 TI - Responses of circulating fish phagocytes to paper mill effluent exposure. PMID- 9871005 TI - Root growth inhibition, photosynthetic pigments production, and metal accumulation in Sinapis alba as the parameters for trace metals effect determination. PMID- 9871007 TI - Acute lethal toxicity of the organophosphorus pesticide chlorpyrifos to different species and strains of Artemia. PMID- 9871006 TI - Influence of petroleum at a refinery terminal on feral winter flounder, Pleuronectes americanus. PMID- 9871008 TI - Use of Mfu-galactoside enzymatic activity as ecotoxicological endpoint on rainbow trout red blood cells. PMID- 9871009 TI - Cadmium accumulation and metallothionein biosynthesis in Cyprinus carpio tissues. PMID- 9871010 TI - Nitrite-induced cell proliferation and polyamine synthesis in the small intestine of mice. PMID- 9871012 TI - The anaerobic degradation of L-serine and L-threonine in enterobacteria: networks of pathways and regulatory signals. AB - The mechanisms controlling the biosynthesis and degradation of l-serine and l threonine are remarkably complex. Their metabolism forms a network of pathways linking several amino acids, central primary metabolites such as pyruvate, oxaloacetate and 3-phosphoglycerate, and C1 metabolism. Studies on the degradation of these amino acids in Escherichia coli have revealed the involvement of fascinating enzymes that utilise quite diverse catalytic mechanisms. Moreover, it is emerging that both environmental and metabolic signals have a major impact in controlling enzyme synthesis This is exemplified by the anaerobically regulated tdc operon, which encodes a metabolic pathway for the degradation of serine and threonine. Studies on this pathway are beginning to provide insights into how an organism adapts its genetic makeup to meet the physiological demands of the cell. PMID- 9871013 TI - Activation of the H+-ATPase in the plasma membrane of cells of Saccharomyces cerevisiae grown under mild copper stress. AB - Cells of Saccharomyces cerevisiae exhibited a more active plasma membrane H+ ATPase during growth in media supplemented with CuSO4 concentrations equal to or below 1 mM than did cells cultivated in the absence of copper stress. Maximal specific activities were found with 0.5 mM CuSO4. ATPase activity declined when cells were grown with higher concentrations up to 1.5 mM (the maximal concentration that allowed growth), probably due to severe disorganization of plasma membrane. Cu2+-induced maximal activation was reflected in an increase of Vmax (approximately threefold) and in the slight decrease of the Km for MgATP (from 0.93 +/- 0.13 to 0.65 +/- 0.16 mM). The expression of the gene encoding the essential plasma membrane ATPase (PMA1) was reduced with a dose-dependent pattern in cells grown with Inhibitory concentrations of copper, while the weakly expressed PMA2 gene promoter was moderately more efficient in cells cultivated under mild copper stress (1.5-fold maximal activation). ATPase was activated by copper despite the slightly lower content of ATPase protein in the plasma membrane of Cu2+-grown cells and the powerful inhibitory effect of Cu2+ in vitro. PMID- 9871016 TI - Studies of the production and characterization of laccase activity in the basidiomycete coriolopsis gallica, an efficient decolorizer of alkaline effluents PMID- 9871014 TI - Effect of high sugar concentration on nitrogenase activity of Acetobacter diazotrophicus. AB - Acetobacter diazotrophicus is a nitrogen-fixing bacterium that growth inside sugar cane plant tissue where the sucrose concentration is approximately 10%. The influence of high sugar content on nitrogenase was measured in the presence of oxygen and of nitrogen added in the form of ammonium and amino acids. In all parameters analyzed, 10% sucrose protected nitrogenase against inhibition by oxygen, ammonium, some amino acids, and also to some extent by salt stress. The oxygen concentration at which inhibition occurred increased from 2 kPa in 1% glucose or gluconic acid, to 4 kPa (0.4 atm) in 10% sucrose. Nitrogenase activity was partially inhibited by increased ammonium levels (2.0, 5.0, and 10.0 mM) in the presence of 1% sucrose, but the cells maintained their nitrogenase activity at 10% sucrose. This could be explained by the slow ammonium assimilation by the cells in the presence of high sucrose concentrations, i.e., independent of its concentration between 2 and 10 mM, the assimilation of ammonium was reduced to one-third in cells grown with 10% sucrose. Some amino acids were also tested in the presence of 1 and 10% sucrose. Cells grown in 1% sucrose had their nitrogenase activity reduced by 50-98% in the presence of glutamic acid, glutamine, alanine, asparagine, or threonine, whereas with 10% sucrose, nitrogenase activity was increased by glutamic acid and was reduced by only 61 73% by the other amino acids. The effect of NaCl concentrations (0.0, 0.25, 0.5, 0.75, or 1.0%) was also studied at the two concentrations of sucrose. Nitrogenase activity and growth of A. diazotrophicus, which was visualized by the pellicle formation in semi-solid medium, showed sensitivity even to low NaCl concentrations, which was somewhat relieved at the higher sucrose level. These observations indicate different osmotolerance mechanisms for sucrose and salt. PMID- 9871017 TI - Purification and characterization of eugenol dehydrogenase from pseudomonas fluorescens E118 PMID- 9871015 TI - Bacillus arsenicoselenatis, sp. nov., and Bacillus selenitireducens, sp. nov.: two haloalkaliphiles from Mono Lake, California that respire oxyanions of selenium and arsenic. AB - Two gram-positive anaerobic bacteria (strains E1H and MLS10) were isolated from the anoxic muds of Mono Lake, California, an alkaline, hypersaline, arsenic-rich water body. Both grew by dissimilatory reduction of As(V) to As(III) with the concomitant oxidation of lactate to acetate plus CO2. Bacillus arsenicoselenatis (strain E1H) is a spore-forming rod that also grew by dissimilatory reduction of Se(VI) to Se(IV). Bacillus selenitireducens (strain MLS10) is a short, non-spore forming rod that grew by dissimilatory reduction of Se(IV) to Se(0). When the two isolates were cocultured, a complete reduction of Se(VI) to Se(0) was achieved. Both isolates are alkaliphiles and had optimal specific growth rates in the pH range of 8.5-10. Strain E1H had a salinity optimum at 60 g l-1 NaCl, while strain MLS10 had optimal growth at lower salinities (24-60 g l-1 NaCl). Both strains have limited abilities to grow with electron donors and acceptors other than those given above. Strain MLS10 demonstrated weak growth as a microaerophile and was also capable of fermentative growth on glucose, while strain E1H is a strict anaerobe. Comparative 16S rRNA gene sequence analysis placed the two isolates with other Bacillus spp. in the low G+C gram-positive group of bacteria. PMID- 9871018 TI - Oxidation of 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT) by Alcaligenes eutrophus A5. AB - Previous studies demonstrated that Alcaligenes eutrophus A5 transforms 1,1,1 trichloro-2,2-bis(4-chlorophenyl)ethane (DDT) to 4-chlorobenzoate via a meta-ring fission product. The initial reactions could be catalyzed by either monooxygenase or dioxygenase enzymes. In the present study, a transient intermediate that accumulated during the transformation of DDT by the biphenyl-grown cells was identified as 1,1,1-trichloro-2-(4-chlorophenyl-2,3-dihydro-4,6-cyclohexadiene)-2 (4'- chlorophenyl)ethane (DDT-2,3-dihydrodiol) on the basis of mass spectral analysis after n-butylboronic acid derivatization. The dihydrodiol undergoes a characteristic acid-catalyzed dehydration to produce phenols. 1H-NMR indicated a cis-relative stereochemistry. The results indicate that the biphenyl dioxygenase from A. eutrophus A5 catalyzes the dihydroxylation of DDT at the unsubstituted carbons on the aromatic ring to produce DDT-2,3-dihydrodiol. PMID- 9871019 TI - Inhibition of colicin synthesis by the antibiotic globomycin. AB - The antibiotic globomycin, an inhibitor of LspA (the lipoprotein signal peptidase), inhibited synthesis of colicin by Escherichia coli cells grown in rich medium. This inhibition was stronger in cells with mutation(s) within either the colicin operon, which is located on a plasmid, or the host chromosome. This phenotype was called Gbc (globomycin blocks colicin synthesis). The Gbc phenotype was affected by growth conditions since it was partially or totally suppressed in cells subjected to high temperatures, treated with sodium azide, or grown in minimal medium. The Gbc phenotype observed with colicin-A-producing cells was more severe in strains carrying plasmids with a deletion within caa (the first gene of the colicin A operon), which encodes colicin A, than in cells with the wild-type caa gene. The Gbc phenotype was alleviated by a null mutation in the degP gene encoding the DegP/HtrA protease, abolished by a null mutation in the lpp gene encoding the murein-lipoprotein, and enhanced by a mutation in the pldA gene encoding the outer membrane phospholipase A. Transcription of the colicin A operon was blocked in cells exhibiting the Gbc phenotype as evidenced by rifampicin treatment of induced cells. This phenotype suggests that either a lipoprotein or a protein involved in lipoprotein metabolism might be involved in the regulation of the expression of the colicin operons and that the colicin A structural gene might play a role in the regulation of transcription of the colicin A operon. PMID- 9871020 TI - Isolation and characterization of enterocin EJ97, a bacteriocin produced by Enterococcus faecalis EJ97. AB - The bacteriocinogenic strain of Enterococcus faecalis EJ97 has been isolated from municipal waste water. It produces a cationic bacteriocin (enterocin EJ97) of low molecular mass (5,340 Da) that is very stable under mild heat conditions and is sensitive to proteolytic enzymes. The amino acid sequence of the first 18 N terminal residues of enterocin EJ97 indicates that it is different from other known protein sequences. Enterocin EJ97 is active on several gram-positive bacteria including enterococci, several species of Bacillus, Listeria, and Staphylococcus aureus. The producer strain is immune to bacteriocin. Enterocin EJ97 has a concentration-dependent bactericidal and bacteriolytic effect on E. faecalis S-47. PMID- 9871021 TI - Biosynthesis of cobalamin in Salmonella typhimurium: transformation of riboflavin into the 5,6-dimethylbenzimidazole moiety. AB - In order to elucidate the biosynthesis of the base moiety of cobalamin in Salmonella typhimurium LT2, this organism was grown in the presence of [1' 14C]riboflavin. The vitamin B12 isolated was 14C-labeled. It was shown by chemical degradation that the 14C-label was exclusively localized in carbon atom 2 of the 5,6-dimethylbenzimidazole moiety. This demonstrated the precursor function of riboflavin in the biosynthesis of 5,6-dimethylbenzimidazole in S. typhimurium. PMID- 9871022 TI - Effect of oxidative stress on the biosynthesis of 2-C-methyl-D-erythritol-2,4 cyclopyrophosphate and isoprenoids by several bacterial strains. AB - In this study, the gram-negative bacteria Xanthomonas campestris, Xanthomonas maltophilia, and Pseudomonas putida, facultative parasites of plants and animals, were shown to accumulate 2-C-methyl-d-erythritol-2,4-cyclopyrophosphate (MEC) in response to benzyl-viologen-induced oxidative stress. Corynebacterium ammoniagenes mutants capable of accumulating MEC in the absence of an exogenous oxidative stress inducer were obtained. Isoprenoid synthesis and MEC synthesis in these and other bacteria were shown to be alternative processes, while biosynthesis of brominated polyene xanthomonadin (an antioxidant pigment of X. campestris) increased concomitantly with the accumulation of MEC. PMID- 9871092 TI - 123I-5-I-R91150, a new single-photon emission tomography ligand for 5-HT2A receptors: influence of age and gender in healthy subjects. AB - 5-HT2A receptors have been implicated in the pathophysiology of mood disorders and in the therapeutic effect of the so-called atypical antipsychotics. Recently, a new radioiodinated ligand with high affinity and selectivity for serotonin 5 HT2A receptors, 123iodinated 4-amino-N-1-[3-(4-fluorophenoxy)propyl]-4-methyl-4 piperidinyl] 5-iodo-2-methoxybenzamide (123I-5-I-R91150), has been developed and has been shown to be suitable for single-photon emission tomography (SPET) imaging. In this study the influence of age and gender on the ligand binding was investigated in normal volunteers. One hundred and fifty MBq of 123I-5-I-R91150 was administered to 26 normal volunteers (13 females and 13 males) with an age range of 23-60 years. SPET imaging was performed with a triple-headed gamma camera. For semi-quantitative analysis, ratios of ligand binding in different regions of interest to the binding in the cerebellum were calculated. Mean ratios of 1.7 were obtained. No gender difference was demonstrated. 5-HT2A binding was shown to decline with age. Over an age range of 40 years a reduction in ligand binding of 42% +/- 7% was found. These results are in agreement w in vitro and positron emission tomography findings of a decline in 5-HT2A receptor binding with age. The findings confirm the suitability of 123I-5-I-R91150 for SPET imaging of 5-HT2A receptors, and highlight the necessity for age-matched controls in clinical studies. PMID- 9871093 TI - Estimation of regional lung function in interstitial pulmonary disease using 99mTc-technegas and 99mTc-macroaggregated albumin single-photon emission tomography. AB - For quantitative evaluation of the regional lung function in patients with interstitial pulmonary disease (IP) in the sitting position, 99mTc-Technegas and 99mTc-macroaggregated albumin (MAA) single-photon emission tomography (SPET) studies were performed in 12 healthy controls (HC) and 42 IP patients. Four transverse images were prepared from the data obtained and designated as slices no. 1-4 from the top downward. Regions of interest (ROIs) were determined in the anterior and posterior parts of the lung in each slice, and the ratio of the count per voxel in the ROIs to the count in the entire lung was calculated as the regional Technegas index (T). The regional perfusion index (Q) was calculated by a similar procedure using the data of 99mTc-MAA SPET. The ratios between T and Q (T/Q) in the anterior and posterior regions of the lung, and the ratios of T and Q between the anterior and posterior regions of the lung (Tp/Ta and Qp/Qa) were examined. In the HC group, T/Q decreased but Tp/Ta and Qp/Qa increased from the upper to the lower lung fields. When IP patients were classified into (I) those in whom T/Q decreased from the upper to the lower lung fields, (II) those in whom it was similar in all slices, (III) those in whom it increased from slice 3 to slice 4, and (IV) those in whom it increased from slice 2 to slices 3 and 4, this classification was more closely correlated with %VC than with %DLCO or PaO2. When the patients were classified according to Tp/Ta and Qp/Qa into (A) those in whom the values were greater in the lower than the upper lung field, (B) those in whom the values were similar in all slices, and (C) those in whom the values were smaller in lower than in upper lung fields, categories B and C were observed frequently even in patients whose %VC was in the normal range. This method is considered to be an effective means to evaluate the progression and pathology of IP and to detect early impairment of lung function. PMID- 9871094 TI - Evaluation of renal function in low-dose cyclosporine-treated patients using technetium-99m diaminocyclohexane: a cationic tubular excretion agent. AB - Technetium-99m diaminocyclohexane (DACH) is a new tubular agent excreted via a cationic transport mechanism, like cyclosporine-A (CsA). It is expected that 99mTc-DACH will permit effective assessment of tubular function in CsA-treated patients. To establish the pharmacokinetic characteristics of 99mTc-DACH and to ascertain whether this new agent is useful in CsA-treated patients, 11 healthy volunteers and 15 CsA-treated patients underwent renal imaging and clearance studies using 99mTc-DACH and chromium-51 ethylene diamine tetra-acetic acid (EDTA). 99mTc-DACH yielded satisfactory dynamic renal images in all participants. The mean plasma clearance of 99mTc-DACH was significantly greater than that of 51Cr-EDTA in volunteers (109.4?19.7 ml/min versus 86.6 +/- 13.7 ml/min, P<0.05). However, the urinary excretion of 99mTc-DACH at 90 min was significantly lower than that of 51Cr-EDTA (46.1% +/- 9.3% versus 53.1% +/- 8.6%, P<0.05), most probably due to its partial parenchymal retention. The elimination half-life of 99mTc-DACH was significantly increased in CsA-treated patients in comparison to volunteers, and consequently the plasma clearance values were significantly suppressed in these patients, in contrast to 51Cr-EDTA and endogenous creatinine clearance values. In conclusion, our findings indicate that 99mTc-DACH, as a sensitive marker of cationic tubular function, could be used to monitor renal haemodynamics in patients receiving CsA treatment. PMID- 9871095 TI - Use of 123I-IMP brain SPET to predict outcome following STA-MCA bypass surgery: cerebral blood flow but not vasoreactivity is a predictive parameter. AB - Superficial temporal artery - middle cerebral artery (STA-MCA) bypass surgery might improve the cerebral blood flow (CBF) but fail to reduce the risk of post surgical events such as ischaemic stroke. In this study, we studied retrospectively whether the risk of post-surgical events corresponded to the change in resting CBF and/or the change in vasoreactivity observed before and after STA-MCA surgery using N-isopropyl-I-123-p-iodoamphetamine (123I-IMP) brain single-photon emission tomography (SPET). 123I-IMP brain SPET images obtained at rest and following acetazolamide challenge both before and after STA-MCA surgery were studied in 30 patients. Resting CBF was estimated semiquantitatively using the resting count ratios of the middle cerebral artery (MCA) area to the cerebellum. Acetazolamide c hallenge was assumed to result in an average increase in flow of 40% in the cerebellum. The vasoreactive response was then estimated as the ratio of the change in counts (acetazolamide - rest) to the resting cerebellar counts multiplied by 1.4. We classified 14 patients (50.5 +/- 19.3 years) into group I, who h a change in their resting CBF from before to after surgery of >10%, and 16 patients (54.0 +/- 18.8 years) into group II, who had a change in their resting CBF from before to after surgery of ?leq10%. Oxygen-15 positron emission tomography (PET) studies were performed in five patients from group I and five patients from group II. One post-surgical event occurred in group I while there were eight post-surgical events in group II (P<0.05). Resting CBF improved by 20.4% +/- 7.5% in group I and by 0.9% +/- 6.9% in group II patients after surgery (P<0.001). No significant difference in the improvement in vasoreactivity was observed between group I patients (32.6% +/- 17.7%) and group II patients (24.6% +/- 15.6%) following surgery. Patients in the group I PET subset showed normalization of oxygen extraction fraction (OEF) from 0.59 +/- 0.09 before surgery to 0.46 +/- 0.06 after surgery (P<0.05), while patients in the group II PET subset showed no statistical difference in OEF before surgery (0.58 +/- 0.14) and after surgery (0.54 +/- 0.05). We conclude that the outcome of STA-MCA bypass surgery can be predicted by the improvement in resting CBF but not by the improvement in vasoreactivity. PMID- 9871096 TI - Effects of aldose reductase inhibitor and vitamin B12 on myocardial uptake of iodine-123 metaiodobenzylguanidine in patients with non-insulin-dependent diabetes mellitus. AB - This study was undertaken to examine the effects of aldose reductase inhibitor (ARI) and vitamin B12 (VB12) on myocardial uptake of iodine-123 metaiodobenzylguanidine (MIBG) in patients with diabetic autonomic disorder. Myocardial scintigraphy using 123I-MIBG was performed on 20 healthy volunteers (controls) and 56 patients with non-insulin-dependent diabetes mellitus (NIDDM), in order to obtain the heart/mediastinum ratio in the initial (HMi) and the delayed images (HMd), and the washout rate (%WR). Thirty-four of the 56 NIDDM patients could be diagnosed as having diabetic autonomic disorder by evaluating their scintigraphic findings in comparison with the controls. Seventeen of these 34 patients received 150 mg/day of doses before meals, and the other 17 received 1.5 mg/day of mecobalamin (VB12 group) in three divided doses after meals, for 3 5 months. According to the presence or absence of clinical symptoms of autonomic or peripheral somatic nerve disorder, the patients were subclassified into four groups. group 1=patients, with autonomic symptoms or somatosensory disorder in the ARI group; group 2=patients without autonomic symptoms or somatosensory disorder in the ARI group; group 3=patients with autonomic symptoms or somatosensory disorder in the VB12 group; and group 4=patients without autonomic symptoms or somatosensory disorder in the VB12 group. After completion of the treatment, myocardial scintigraphy was performed again. Comparing the results obtained before and after the treatment, it was seen that ARI improved only the HMi in group 1 (P=0.046), whereas VB12 significantly improved HMi in the group 3 (P=0.018) and HMi, HMd and %WR in group 4 (P=0.043, P=0.018 and P=0.043, respectively). We conclude that VB12 is more efficacious than ARI in the treatment of diabetic cardiovascular autonomic disorder. PMID- 9871098 TI - Efficient scatter modelling for incorporation in maximum likelihood reconstruction. AB - Definition of a simplified model of scatter which can be incorporated in maximum likelihood reconstruction for single-photon emission tomography (SPET) continues to be appealing; however, implementation must be efficient for it to be clinically applicable. In this paper an efficient algorithm for scatter estimation is described in which the spatial scatter distribution is implemented as a spatially invariant convolution for points of constant depth in tissue. The scatter estimate is weighted by a space-dependent build-up factor based on the measured attenuation in tissue. Monte Carlo simulation of a realistic thorax phantom was used to validate this approach. Further efficiency was introduced by estimating scatter once after a small number of iterations using the ordered subsets expectation maximisation (OSEM) reconstruction algorithm. The scatter estimate was incorporated as a constant term in subsequent iterations rather than modifying the scatter estimate each iteration. Monte Carlo simulation was used to demonstrate that the scatter estimate does not change significantly provided at least two iterations OSEM reconstruction, subset size 8, is used. Complete scatter-corrected reconstruction of 64 projections of 40?128 pixels was achieved in 38 min using a Sun Sparc20 computer. PMID- 9871097 TI - Technetium-99m sestamibi brain single-photon emission tomography for detection of recurrent gliomas after radiation therapy. AB - Technetium-99m sestamibi (MIBI), an alternative radiopharmaceutical for myocardial perfusion imaging, has also been proposed for use as an imaging agent for various tumours, including breast cancer, lung cancer, lymphomas, melanomas and brain tumours. After routine radiation therapy, deteriorating clinical status or treatment failure may be due to either radiation-induced changes or recurrent tumour. Computed tomography and magnetic resonance imaging offer imperfect discrimination of tumour viability and radionecrosis. Against this background we undertook a retrospective study of 35 malignant glioma patients in whom clinical deterioration had occurred, in order to clarify the value of 99mTc-MIBI SPET in identifying tumour recurrence. SPET was performed 15 min after intravenous injection of 1110 MBq with a dual-headed gamma camera using a fan-beam collimator. Transverse, coronal and sagittal views were reconstructed. Intense MIBI uptake was found in 31 patients. This uptake was correlated with tumour recurrence as proved by histology and/or rapid, fatal evolution of these cases. The statistical analysis performed on this population of patients with MIBI uptake revealed a group of patients with a long mean survival and a group with a short mean survival. Two subgroups were found within each of these groups, according to the functional index ratio (tumour uptake/pituitary gland uptake ratio). No MIBI uptake was found in four patients who are still alive and can be considered to be disease-free. In those cases showing MIBI uptake, death occurred an average of 6.69 months following brain SPET. According to our results, the specificity and sensitivity of 99mTc-MIBI brain SPET seem to be high. Moreover, this technique is more accurate than computed tomography or magnetic resonance imaging for discriminating between tumour recurrence and radionecrosis. PMID- 9871100 TI - Highlights lecture of the European Association of Nuclear Medicine and the World Federation of Nuclear Medicine and Biology Congress, Berlin 1998. Where next and how? AB - Nuclear medicine benefits not only the people of developed countries but also those who are in developing or deteriorating countries. The combination of diagnostic imaging, tissue characterisation, function measurement and targeted therapy is powerful and cost-effective. This congress provides a sample of nuclear medicine's contribution to the world. PMID- 9871099 TI - Iodine-123 labelled nor-beta-CIT binds to the serotonin transporter in vivo as assessed by biodistribution studies in rats. AB - Iodine-123 labelled 2beta-carbomethoxy-3beta-4-iodophenylnortropane (nor-beta CIT), a radioiodinated cocaine analogue, was evaluated as an agent for the in vivo labelling of serotonin transporters by biodistribution studies in rats. Intravenous injection of [123I]nor-beta-CIT resulted in high accumulation of radioactivity in brain areas with high densities of serotonin (hypothalamus) and dopamine transporters (striatum), although the binding was less pronounced in the hypothalamus. While binding of [123I]nor-beta-CIT in the hypothalamus was blocked significantly by fluvoxamine (a selective serotonin transporter blocker) but not by GBR12,909 (a selective dopamine transporter blocker), the opposite was observed in the striatum. The results of this study indicate that [123I]nor-beta CIT, although not being a selective radioligand, binds specifically to serotonin transporters in the hypothalamus in vivo and thus suggest that [123I]nor-beta-CIT promises to be a suitable radioligand for single-photon emission tomography imaging of serotonin transporters in humans. PMID- 9871103 TI - Characterization of porins isolated from the outer membrane of Serratia liquefaciens. AB - A major outer membrane protein with an apparent molecular weight of 42 kDa was purified from Serratia liquefaciens grown on Brain Heart Infusion medium. The same protein was obtained when the cells were grown on a synthetic medium supplemented with 2% glucose. The amino acid composition of this protein revealed it to be hydrophilic. The pore-forming ability of the 42-kDa protein was determined by the liposome swelling assay. This assay demonstrated that the protein forms nonspecific channels with a diameter between 1.16 and 1.6 nm. An additional protein with a molecular weight of 47 kDa was obtained on synthetic medium supplemented with maltose. This protein exhibited specific pore-forming ability to maltose and maltodextrins, but was also permeable to other compounds, according to their size. When bacteria were grown on Nutrient Broth medium, two outer membrane proteins with molecular weights of 41 kDa and 42 kDa were produced by the bacteria. All three types of proteins represent monomers of respective oligomers. The monomers did not exhibit pore-forming ability when incorporated into liposomes. We, therefore, propose that the oligomer is the functional unit of a porin capable of forming permeability channels in the outer membrane of Serratia liquefaciens. These results indicate that S. liquefaciens contains several porins exhibiting specific osmoregulation or that are induced by a specific nutrient, where the 42-kDa outer membrane protein of this bacterium is certainly a major porin. PMID- 9871104 TI - The integration host factor (IHF) affects the expression of the phosphate-binding protein and of alkaline phosphatase in Escherichia coli. AB - The genes encoding alkaline phosphatase (phoA) and the inducible inorganic phosphate transport system Pst (pstS,C,A,B,U) belong to the PHO regulon. Mutants of Escherichia coli lacking the global regulatory protein integration host factor (IHF) show an increased level of alkaline phosphatase and a decreased level of Pst. IHF binds weakly but specifically to a DNA fragment containing the promoter region of the pst operon but does not bind to a fragment that includes the promoter region of phoA. It is proposed that IHF is a positive regulator of the pst operon and as such controls indirectly the expression of phoA. PMID- 9871105 TI - In vivo and In vitro function of the intracellular proteolytic apparatus in nongrowing bacillus megaterium under heat stress AB - In Bacillus megaterium sporulating at 35 degreesC, up to 90% of 10-min pulse labeled proteins were degraded. Degradation proceeded in two waves. Short-lived proteins, i.e., intrinsically labile proteins and proteins made short-lived because of starvation, were mostly degraded during the reversible sporulation phase. Their amount corresponded to 20% or slightly more during 2 h. The second wave of protein degradation, which followed during the irreversible sporulation phase at 35 degreesC, increased the amount of total degradable pulse-labeled proteins to about 90%. This wave was absent in the isogenic asporogenic mutant 27 36 or in the wild strain, whose sporulation was inhibited by increased temperature. The proportion of degradable proteins was thus reduced to less than 40% in the asporogenic mutant incubated at 35 degreesC and to 46% in the wild strain whose sporulation was suppressed by the temperature of 47 degreesC. Unlike sporulating cells, these cells were thus capable of degrading short-lived and denatured proteins, but were not able to degrade most of other proteins. The in vitro protein degradation was substantially enhanced by increasing the Ca2+ concentration, suggesting a role of Ca2+-dependent proteinase(s) in the process. PMID- 9871106 TI - Regulation of lipA gene expression by cell surface proteins in arthrobacter photogonimos AB - Expression of the light-inducible lipA gene in Arthrobacter photogonimos by photodynamic compounds and visible light was inhibited by washing cells with 1 M KCl. Addition of cell surface extract to KCl-washed cells restored the induction. Washing cells with 1 M MgCl2 removed a 14-kDa polypeptide and concomitantly caused expression of lipA gene without photodynamic treatment. The purified 14 kDa polypeptide inhibited photodynamic induction of lipA gene. These results indicated that regulation of lipA gene expression occurred at the cell surface and involved positive and negative factors, probably through a signal transduction system. PMID- 9871107 TI - Evidence for Clostridium perfringens enterotoxin (CPE) inducing a mitogenic and cytokine response in vitro and a cytokine response in vivo. AB - We investigated some immunogenic properties of Clostridium perfringens enterotoxin (CPE) in vitro using murine J774A macrophages (MPhi) and in vivo using Swiss Webster (SW) mice. CPE was a potent mitogen in vitro, where cell proliferation increased with CPE concentration. CPE was nonmitogenic when MPhi were concurrently incubated with CPE and interferon gamma (IFN-gamma). MPhi incubated in the presence of CPE induced the synthesis of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma), but not interleukin-2 (IL-2). In vivo, CPE induced a pro inflammatory cytokine response with striking production of IFN-gamma, IL-1, and IL-6. Regardless of route of CPE entry, serum cytokine levels generally peaked within 1 h of administration and were maintained for 4-8 h. Although CPE engenders an intense immune response during toxicosis, the toxin does not appear to be a superantigen. Death from CPE-induced shock appears to result from various interrelating immunological mechanisms. PMID- 9871108 TI - Alpha-galactosidase of Bifidobacterium adolescentis DSM 20083. AB - Bifidobacterium adolescentis was grown anaerobically in medium enriched with alpha-D-galactosides. alpha-Galactosidase (EC 3.2.1. 22) was released from the cells by ultrasonic treatment and purified 36-fold by ultrafiltration, ammonium sulphate precipitation, anion-exchange chromatography, and size-exclusion chromatography. Two protein bands were consistantly observed after sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). Electrophoretically homogeneous alpha-galactosidase was only obtained by electroelution. The enzyme had an apparent molecular mass of 344 kDa and 79 kDa as judged by size-exclusion chromatography and SDS-PAGE, respectively. Activity-staining after nondenaturing SDS-PAGE indicated an apparent molecular mass of 145 kDa. Thus, a tetrameric structure of the protein is suggested. The alpha-galactosidase showed optimal activity at pH 5.5 and 55 degrees C. Lower pH values and higher temperatures rapidly inactivated alpha-galactosidase. The enzyme hydrolyzed specifically alpha galactosidic linkages, and alpha-(1-3)-linkages were hydrolyzed at a higher rate compared to alpha-(1-6)-linkages. Hydrolysis of galactosides followed normal saturation kinetics; KM-values for p-nitrophenyl-alpha-galactopyranoside (p-NPG) and raffinose were calculated with 0.957 mM and 4.12 mM, respectively. PMID- 9871109 TI - Sequence analysis of small cryptic plasmids isolated from Selenomonas ruminantium S20. AB - Two small cryptic plasmids designated pONE429 and pONE430 were isolated from a rumen bacterium, Selenomonas ruminantium S20. The complete sequence of pONE429 was 2100 bp and contained one open reading frame (ORF) of 201 amino acids. The sequence of pONE430 had 1527 bp and one ORF of 171 amino acids with the similarity of replication protein (Rep protein) of pOM1, pSN2, and pIM13 isolated from Butyrivibrio fibrisolvens, Staphylococcus aureus, and Bacillus subtilis, respectively. In these plasmids, the upstream nucleotide sequence of Rep protein had the conserved nucleotides which could be double-strand origin (DSO) of rolling circle replication (RCR) mechanism. The plasmids of pONE429, pONE430, pJJMI, pJDB21, and pS23 were isolated from S. ruminantium strains and had similar regions that were located within a <450-bp nucleotide. These similar regions may be the location that was recognized by the host strain, S. ruminantium. PMID- 9871110 TI - Regulation of the Escherichia coli secA gene is mediated by two distinct RNA structural conformations. AB - Expression from the secA gene, encoding a key component of the general secretory pathway of Escherichia coli, is influenced by the secretion status of the cell, autogenous translational repression, and translational coupling to the upstream gene, X. SecA binds to its mRNA in a region overlapping its ribosome binding site, thus competing with ribosomes that would initiate secA translation. Mapping of the geneX-secA mRNA secondary structure has demonstrated that the RNA can adopt two distinct conformations in solution. The first conformation arises from the base-pairing of the secA Shine-Dalgarno (SD) sequence with the geneX terminus. The second conformation, in which the secA SD sequence is no longer paired with the geneX terminus, contains a GC-rich stem upstream of the secA SD sequence. The presence of this GC-rich stem is supported by structure mapping of a mutant RNA containing a deletion in the geneX terminus. The former structure appears to be involved in translational coupling by directly linking the geneX and secA sequences, where geneX translation activates secA translational initiation through the unpairing and unmasking of the secA SD sequence. As indicated by SecA-RNA binding assays, the latter structure is probably involved in SecA binding and translational repression of the secA gene. The stabilizing effect of magnesium ions toward occlusion of the secA SD sequence supports the presence of RNA tertiary structure in this regulatory domain. PMID- 9871111 TI - The autonomously replicating sequence (ARS) of the yeast Saccharomyces exiguus Yp74L-3. AB - Fragments containing ARSes were cloned from the genomic DNA of the yeast Saccharomyces exiguus Yp74L-3, and the essential regions for ARSes were restricted for these fragments. Mapping studies of ARS-acting sequences in one of these fragments suggested that S. exiguus recognizes a sequence as an ARS that is different from that recognized by Saccharomyces cerevisiae. Two ARS essential regions of S. exiguus were sequenced, and an ARS core consensus sequence of S. exiguus was deduced to be MATTAMWAWWTK. This sequence differs significantly from that of S. cerevisiae in two positions, suggesting that these nucleotide substitutions cause the difference in the ARS-recognition modes between S. exiguus and S. cerevisiae. PMID- 9871112 TI - Identification of the high-virulence clone of group B streptococci in Mexican isolates by growth characteristics at 40 degrees C. AB - Group B streptococci (GBS) colonizing the vagina and rectum of pregnant women cause invasive disease of the offspring in a small number of cases. The immune status of the host and differences in virulence among strains appear to be the main determinants for neonatal infection. A high-virulence clone (HVC) was proposed to cause much of the morbidity and mortality when a collection of GBS isolates was examined by multilocus enzyme electrophoresis. HVC isolates could be further distinguished by their inability to grow at 40 degrees C. This characteristic was used in the present study to examine a collection of 57 GBS isolates from Mexico City for the HVC. Three serotype III invasive strains were classified in the HVC. The other eleven invasive strains and all carrier isolates had growth curves unaffected at 40 degrees C. These results demonstrate the presence of the HVC in Mexico. Such a low prevalence could explain in part the low rate of GBS invasive neonatal disease in Mexico. PMID- 9871113 TI - Comparison of methods to enumerate bacteria in dental unit water lines. AB - Millipore HPC samplers are simple, self-contained test devices that can be used by personnel in dental offices who do not have microbiologic training to easily and economically monitor dental unit water quality without laboratory support. This study evaluated the correlation of HPC samplers to R2A agar for enumerating planktonic bacteria in dental unit treatment water. Eight different dental units were sampled. Five replicates were performed for each media at each dilution. The Pearson correlation coefficient between the R2A agar and HPC sampler is 0.89. These data suggest HPC samplers correlate with conventional laboratory-based R2A culture techniques for determining dental unit water line contamination. PMID- 9871114 TI - Detection of messenger RNA transcribed from genes encoding enzymes of amino acid biosynthesis in Buchnera aphidicola (endosymbiont of aphids). AB - The aphid Schizaphis graminum is dependent on Buchnera aphidicola, a prokaryotic endosymbiont. One of the functions of the endosymbiont is the synthesis of essential amino acids for the aphid host. Previously we have found that B. aphidicola has many of the genes that encode enzymes of amino acid biosynthesis. Using reverse transcriptase and the polymerase chain reaction, we have detected messenger RNA corresponding to genes involved in the synthesis of tryptophan, isoleucine, valine, leucine, and histidine. PMID- 9871115 TI - Schizosaccharomyces pombe exo1 is involved in the same mismatch repair pathway as msh2 and pms1. AB - Besides the MutLS-like system, Schizosaccharomyces pombe has an additional pathway of mismatch repair. This minor pathway, producing short excision tracts, repairs C/C and, with lower efficiency, other mismatches also. We investigated the involvement of the exo1+, msh2+ and pms1+ genes in the two pathways. The exo1+ gene encodes a 5' to 3' exonuclease, while msh2+ and pms1+ are homologs of Escherichia coli mutS and mutL, respectively. Intragenic two-factor crosses showed that exo1+, msh2+ and pms1+ are involved in the major, but not in the C/C correcting, pathway. Post-meiotic segregation frequencies and mitotic mutation rates in single and double mutants supported this finding. Furthermore, msh2 delta was epistatic over exo1 delta, and the ExoI enzyme is likely to be redundant with other exonucleases. PMID- 9871116 TI - Glucose repression on RIM1, a gene encoding a mitochondrial single-stranded DNA binding protein, in Saccharomyces cerevisiae: a possible regulation at pre-mRNA splicing. AB - The mitochondrial single-stranded DNA-binding protein (SSB) encoded by anuclear gene, RIM1, is a homolog of Escherichia coli SSB. The addition of glucose decreased the amount of RIM1-mRNA in cells growing in a glycerol medium, but increased the amount of the immature RIM1-mRNA. The changes in the amounts of both mature and immature RIM1-mRNAs were dependent on SRN1/REG1/HEX2, a gene relating to pre-mRNA-splicing and glucose repression. These observations suggest that the expression of the mitochondrial SSB is regulated, at least in part, by pre-mRNA splicing under the control of glucose repression. PMID- 9871117 TI - Genetic analysis of the yeast NUD1 endo-exonuclease: a role in the repair of DNA double-strand breaks. AB - The deoxyribonucleases (DNases) have been shown genetically to be important in the vital processes of DNA repair and recombination. The NUD1 gene, which codes for an endo-exonuclease of Saccharomyces cerevisiae, was analyzed for its role in the DNA double-strand break (DSB) repair processes. While the nud1 strain is only slightly sensitive to ionizing radiation, expression of the HO-endonuclease to introduce a DSB at the MAT locus in that strain results in cell death. Cell survival is inversely proportional to the duration of HO-endonuclease expression. Analysis of the surviving colonies from the nud1 strain indicated that many of the survivors are sterile and that the proportion of these sterile survivors increases with the time of HO-endonuclease expression. On the other hand, the surviving colonies from the isogenic NUD1 strain are mating-proficient. Interestingly, double mutants of nud1 rad52 are more resistant to ionizing irradiation than the rad52 strain and have a cell-survival fraction of 32% for rad52-1 nud1 and 9% for rad52::URA3 nud1 following prolonged HO-endonuclease expression, indicating that nud1 has a suppressor effect on the DSB-induced lethality in rad52. Polymerase chain reaction analysis showed that many of the nud1 survivors contained small alterations within theMAT locus, suggesting that the survivors arose through the process of non-homologous end-joining. These results suggest that the endo-exonuclease acts at a DSB to promote DNA repair via the homologous recombination pathway. PMID- 9871118 TI - Molecular cloning and genetic characterization of the Saccharomyces cerevisiae NGS1/MRE11 gene. AB - The Saccharomyces cerevisiae ngs1-1 mutant was previously identified by its enhanced sensitivity to simple DNA-alkylating agents such as methyl methanesulfonate but not to UV. Molecular cloning and sequencing of NGS1 as a putative DNA-alkylation repair gene revealed that it isidentical to MRE11, a gene that is involved in DNA recombinational repair. In order to investigate functional domains of the Mre11 protein, nucleotide-sequence alterations of a number of mre11 mutant alleles, including ngs1-1, mre11-1 (ts), mre11-2, mre11-3 and mre11-58, were determined. Most of these mutations map to the N-terminus ofMre11, emphasizing the importance of this highly conserved domain. The ngs1-1 and mre11-3 mutants carry nonsense mutations resulting in truncated proteins. Missense mutations were found in mre11-1 (ts), mre11-2 and mre11-58, of which mre11-2 and mre11-58 mapped to the conserved phosphoesterase domains, indicating the involvement of these motifs in the formation and/or processing of DNA double strand breaks. Finally, mitotic-recombination assays show that the mre11 delta mutation enhances inter-chromosomal recombination but decreases the intra chromosomal deletion frequency. In addition, MRE11 appears to play different roles during spontaneous and alkylation-induced homologous mitotic recombination. PMID- 9871119 TI - Identification and functional analysis of a Kluyveromyces lactis homologue of the SPT4 gene of Saccharomyces cerevisiae. AB - Sequence analysis of a DNA fragment containing the KlCOX18 gene originating from chromosome II of the yeast Kluyveromyces lactis revealed the presence of an adjacent open reading frame (ORF) for a protein exhibiting 78.4% identity with the Saccharomyces cerevisiae Spt4p. Based on the identical length (102 aa) and the conservation of the zinc-finger motif found in Spt4p we named this ORF KlSPT4. When expressed in S. cerevisiae the KlSPT4 gene complemented all spt4 mutant phenotypes. It is proposed that KlSpt4p, like its S. cerevisiae counterpart is a protein involved in the establishment or maintenance of the chromatin structure that influences the expression of many yeast genes. PMID- 9871120 TI - Isolation and characterisation of the acetyl-CoA carboxylase gene from Aspergillus nidulans. AB - The putative gene encoding acetyl-CoA carboxylase, accA, has been isolated from Aspergillus nidulans. This single-copy gene has an open reading frame (ORF) of 6864 bp and contains two small introns near the 5'-end. A short ORF upstream of the ATG start codon has been identified in this gene by RT-PCR. Based on sequence homology to acetyl-CoA carboxylases from other organisms, putative biotin-, ATP-, HCO3-- and acetyl-CoA- binding sites have been assigned. Northern data and ACC enzyme-activity measurements from A. nidulans suggested that expression of accA was higher in media containing nitrate than ammonia as a sole nitrogen source. Deletion of accA in A. nidulans was unsuccessful. The failure of A. nidulans to grow in the presence of the ACC-specific inhibitor, soraphen A, supplemented with C16-18 fatty acids suggested that ACC is an essential enzyme. PMID- 9871121 TI - Isolation of the 3-phosphoglycerate kinase gene of the arbuscular mycorrhizal fungus Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe. AB - The 3-phosphoglycerate kinase (PGK) gene of the arbuscular mycorrhizal fungus Glomus mosseae has been isolated by differential RNA display (DD). Experimentally, the technique of DD was utilized to simultaneously compare the mRNA transcript populations from Lycopersicon esculentum root systems colonized by the arbuscular mycorrhizal fungus G. mosseae and non-mycorrhizal plants. A differentially expressed band was isolated and cloned from mycorrhizal tissue. The isolated DD fragment was screened and sequenced. A reverse transcription-PCR (RT-PCR) technique detected transcripts in germinated spores and G. mosseae colonized root systems but not in uncolonised root systems. The full-length cDNA was isolated by RACE and corresponded to the 3-phosphoglycerate kinase gene of G. mosseae. The cDNA encodes a polypeptide of 416 amino acids, with a predicted molecular weight of 44 764 Da. The PGK proteins shows high homology to those PGKs of other fungi and the phylogenetic relationship among 14 isolated fungal PGK sequences is illustrated. PMID- 9871122 TI - A single amino-acid substitution in the iron-sulphur protein subunit of succinate dehydrogenase determines resistance to carboxin in Mycosphaerella graminicola. AB - A gene encoding the iron-sulphur protein (Ip) subunit of succinate dehydrogenase (Sdh, EC 1.3.99.1) from Mycosphaerella graminicola (Septoria tritici) has been cloned andsequenced. The deduced amino-acid sequence exhibited a high degree of homology to Ip subunits of Sdh from other organisms; three cysteine-rich clusters associated with the iron-sulphur centres involved in electron transport were particularly conserved. Expression studies using a synthetic green fluorescent protein (SGFP) expression vector demonstrated that the cloned DNA also contained a functional promoter region and confirmed that the deduced initiation codon could act as a translational start site. Mutants resistant to the fungicide carboxin (Cbx), a known inhibitor of Sdh, were found to contain a single amino acid substitution in the third cysteine-rich domain of the Ip protein. These mutations resulted in the conversion of a highly conserved His residue, located in a region of the protein associated with the [3Fe-4 S] high-potential non-heme iron sulphur-redox (S3) centre, to either Tyr or Leu. AnIp gene containing the His -> Tyr mutation was constructed and shown to confer Cbx resistance following co-transformation into the Cbx-sensitive wild-type strain. This confirmed that the mutation identified by sequence analysis was responsible for determining Cbx resistance. PMID- 9871123 TI - PCR cloning and detection of point mutations in the eburicol 14alpha-demethylase (CYP51) gene from Erysiphe graminis f. sp. hordei, a "recalcitrant" fungus. AB - Molecular studies of some micro-organisms are hampered by the difficulty of obtaining sufficient amounts of nucleic acids. A cloning strategy based on PCR has therefore been used to clone the eburicol 14alpha-demethylase (CYP51) gene of the obligate fungus Erysiphe graminis f. sp. hordei (Egh) using minute amounts of genomic DNA. The CYP51 gene encodes the enzymatic target of a major group of fungicides. Sequencing CYP51 from different Egh isolates revealed the occurrence of two alleles for this gene. An allele-specific PCR assay was developed to detect each CYP51 allele. PMID- 9871124 TI - Induction of a polyubiquitin gene (ubi1) by potato phytoalexins and heat shock in Gibberella pulicaris. AB - Gibberella pulicaris, a causal agent of potato dry rot, infects potato tubers via wounds, where it is exposed to the phytoalexins rishitin and lubimin. Incubation of mycelium on agar supplemented with phytoalexins transiently induced the transcription of a polyubiquitin gene consisting of four ubiquitin units arranged head to tail; the fourth unit contains a 54-bp intron and an additional glutamine at the C-terminus of the encoded protein. Southern analysis of the G. pulicaris genome revealed one copy of the isolated polyubiquitin gene and one or two copies of other ubiquitin genes. Increased transcription of the gene was detectable above a threshold of 100 microg/ml of rishitin and at elevated temperatures, which indicates that exposure to phytoalexins causes a stress reaction of hyphal cells similar to that after heat shock. PMID- 9871126 TI - Management of intractable intracranial hypertension in severely head-injured patients: second-tier therapy. PMID- 9871127 TI - Lumbar spinal stenosis. PMID- 9871125 TI - Cloning, sequencing and developmental expression of the genes encoding S4 and S10 ribosomal proteins in the cellular slime mould Dictyostelium discoideum. AB - The sequences of the rps4 and rps10 genes encoding the Dictyostelium discoideum homologues of the basic ribosomal proteins S4 and S10 were determined from cDNA and genomic DNA clones. They are expressed respectively as 266 and 153 amino-acid long proteins. In both cases, the N-terminal methionine is cleaved in the mature proteins. S4 contains two putative nuclear targeting signals and displays a strong overall identity (around 60%) to eukaryotic S4 homologues. The rps10 gene harbours a 314-bp intron located close to its 5'-coding end. The overall identity between D. discoideum S10 and eukaryotic homologues is around 38% and rises to 53% in the N-terminal domain. Southern blots suggest that both S4 and S10 are encoded by single genes that are regulated during development. The corresponding mRNAs decrease sharply after 8 h of differentiation. PMID- 9871128 TI - Management of mass lesions of the brain stem. PMID- 9871129 TI - Hydrocephalus chronology in adults: confused state of the terminology. PMID- 9871130 TI - Chemoradiotherapy for pediatric brain tumors. PMID- 9871131 TI - Papers reviewed in this issue. PMID- 9871132 TI - Publications scanned for pertinent articles. PMID- 9871311 TI - Efficacy and mechanisms of action of sodium hypochlorite on Pseudomonas aeruginosa PAO1 phage F116. AB - The Pseudomonas aeruginosa PAO1 phage F116 was used to investigate the viricidal activity and the mechanism of action of sodium hypochlorite. The bacteriophage was inactivated with a low concentration (0.0005% available chlorine) of the biocide prepared in tap water but it was less sensitive to a sodium hypochlorite solution prepared in ultra-pure water (0.0075% available chlorine). For all the effective concentrations of sodium hypochlorite (i.e. producing at least 4 log reduction in phage titre), F116 was readily inactivated within 30 s. Electron microscopical investigations of the phage particles challenged with sodium hypochlorite showed a wide variety of deleterious effects, some of which have not been previously observed with other biocides. The wide range of structural alterations observed suggested that sodium hypochlorite has multiple target sites against F116 bacteriophage. A 30 s exposure to sodium hypochlorite (0.001% available chlorine) produced severe damage, the number and severity of which increased with a higher concentration (0.0075% available chlorine) and with a longer contact time. These observations suggested that sodium hypochlorite inactivated F116 bacteriophage by causing structural alterations to the phage head, tail and overall structure, hence possibly releasing the viral genome from damaged capsids in the surrounding media. PMID- 9871312 TI - Comparison of ribotyping and arbitrarily primed PCR for molecular typing of Salmonella enterica and relationships between strains on the basis of these molecular markers. AB - Arbitrarily primed PCR (AP-PCR) using a discriminatory 10-mer primer and an automated EcoRI ribotyping technique (Riboprinter) were compared for their ability to discriminate between 100 serovars of Salmonella, including multiple isolates representing Salm. Enteritidis PT4 and Salm. Typhimurium DT104. Profiles generated by each method were subjected to numerical analysis using GelCompar software, resulting in the construction of phylogenetic trees and calculation of Simpson's numerical index of diversity (DI). Both methods were highly discriminatory for isolates of Salmonella (Ribotype DI = 0.990, AP-PCR DI = 0.997) with EcoRI ribotyping proving more discriminatory than AP-PCR for isolates of Typhimurium DT104. The population structure was found to be clonal by numerical analysis of markers generated by both methods with serovars being polyphyletic in some cases and grouped in a single cluster in others. No absolute correlation was observed in the relationships between strains formed on the basis of ribo- and AP-PCR markers and serological characteristics. PMID- 9871313 TI - Specific PCR primers from the 16S-23S rRNA spacer region for the rapid detection and identification of Actinobacillus seminis. AB - Actinobacillus seminis is a common cause of ovine epididymitis and ram infertility. The ability to detect and identify this organism promptly is important commercially for the quality control of ram semen samples. Actinobacillus seminis is a fastidious and slow-growing bacterium and primary isolation and presumptive identification can be difficult and time-consuming. In this study, two ribosomal operons, termed rrnA and rrnB, have been characterized in the A. seminis genome, and these contain one and two tRNAs, respectively, in the spacer region between the 16S and 23S rRNA genes. Species-specific primers for A. seminis were developed from the sequence of the spacer region of rrnB for the identification and detection of A. seminis by PCR. The PCR assay was specific for A. seminis and gave no amplification products with phenotypically similar organisms such as Histophilus ovis. Storage solution used to preserve semen for long-term storage was found to inhibit the PCR. Therefore, for diagnostic purposes, the assay would best be performed after primary isolation or perhaps on fresh semen prior to storage if obvious contamination is indicated. PMID- 9871314 TI - Ribotyping and plasmid profiling of Yersinia ruckeri. AB - A total of 183 strains of Yersinia ruckeri, isolated from a wide variety of sources, were studied with respect to their plasmid profile and ribotype patterns. Eight plasmid profiles and 11 ribotypes were demonstrated, with one profile being predominant by both typing methods. The results suggest a clonal structure for this species, with a predominant clone being responsible for most of the outbreaks worldwide. The results of a long-time survey in several fish farms in Spain and Denmark seem to support this idea. PMID- 9871315 TI - Identification and clustering of dairy propionibacteria by RAPD-PCR and CGE-REA methods. AB - A total of 67 classical propionic acid bacteria (PAB) strains, 10 of which were from type culture collections and 57 from milk, typical Italian cheeses, acid whey and feed flour of different regions, were analysed by Randomly Amplified Polymorphic DNA (RAPD-PCR) and by Conventional Gel Electrophoresis Restriction Endonuclease Analysis (CGE-REA). The genotypic traits achieved using RAPD-PCR with three primers (OPL-01, OPL-02 and OPL-05) and SmaI CGE-REA patterns were compared by numerical analysis and allowed a clear distinction of four clusters corresponding to the currently described species of classical propionibacteria according to type and reference strains positions. No discrepancies exist in species recognition between the two methods; 36 isolates were identified as Propionibacterium freudenreichii, 15 as P. jensenii, four as P. acidipropionici and two as P. thoenii. Many differences, however, were observed in intraspecific clustering. Numerical comparison of RAPD-PCR profiles appeared to be a suitable method for highlighting the presence of particular phenotypic characters, while intraspecific differentiation obtained by CGE-REA analysis allowed association of strains at high similarity levels on the basis of their geographical origin. PMID- 9871316 TI - Performance evaluation of disinfectant formulations using poloxamer-hydrogel biofilm-constructs. AB - Poloxamer F127 is a di-block co-polymer of polyoxyethylene and polyoxypropylene. Aqueous solutions show thermo-reversible gelation, being liquid at temperatures < 15 degrees C and robust gels at temperatures > 15 degrees C. Chilled poloxamer solutions (30% w/v) were inoculated with approximately 10(4-5) cfu ml-1 of stationary phase cultures of Pseudomonas aeruginosa, Ps. fluorescens, Pantoea agglomerans, Micrococcus luteus, Staphylococcus epidermidis, Bacillus subtilis or Listeria innocua. Drops (200 microliters) of the inoculated poloxamers were placed on stainless steel coupons held in Petri dishes containing moistened cotton wool and incubated at 30 degrees C for 5 h. All strains grew well giving between 10(6-7) cfu ml-1 at 5-6 h. The cultured gels were readily applied to tests of biocide effectiveness as the stainless steel coupons could be removed and flooded with biocide solution for fixed exposure times. Provided that the temperature of the biocide solutions was > 15 degrees C, the integrity of the gels could be maintained during exposure. After exposure, the gels and their supports were removed to separate tubes containing neutralizer solution (< 15 degrees C). The gels rapidly dispersed within 5 min to ensure a complete recovery of the sample population. Biofilm-constructs and cell suspensions (10(7) cfu ml 1) were exposed to four commercial disinfectant formulations, based on hypochlorite, alcohol, hydrogen peroxide and a tenside, at recommended use levels. Cell suspensions, in the presence of bovine serum albumen (BSA; 0.03% w/v), were subject to a > 5-log kill within 5 min while the killing effected against the biofilm-constructs varied between 0.4 and 2-log reductions. The results indicate a high degree of reproducibility between replicate samples, with patterns of susceptibility varying both as a function of organism, biocide type and concentration. The experiments strongly support the view that poloxamer constructs are suitable for application in trials and testing of disinfectant formulations. PMID- 9871317 TI - Genetic analysis of the bacteriocin-encoding plasmids pRJ6 and pRJ9 of Staphylococcus aureus by transposon mutagenesis and cloning of genes involved in bacteriocin production. AB - pRJ6 and pRJ9, small Staphylococcus aureus plasmids which code for bacteriocins, exhibited a bactericidal activity against several lactic acid bacteria and strains of Listeria monocytogenes, an important food-borne pathogen. Filter mating experiments using plasmid derivatives tagged with either Tn551 or Tn917 lac showed that pRJ6, but not pRJ9, could be mobilized by staphylococcal conjugative plasmids. Transposon mutagenesis of both plasmids was also performed. The bacteriocin and immunity structural genes of pRJ6 are part of the same operon, which is located around co-ordinate 4.0, being transcribed from right to left. However, gene cloning experiments using a staphylococcal vector showed some evidence for the involvement of additional functions of pRJ6 in bacteriocin expression. One function involved in pRJ6 mobilization mapped around co-ordinate 5.2, and it appears to be transcribed from left to right. The bactericidal action exerted by strains harbouring pRJ9 appears to reflect the activity of at least two bacteriocins, whose combined action results in a broader spectrum of activity and in a higher antagonistic activity. Gene cloning experiments also supported these assumptions. PMID- 9871318 TI - The use of poloxamer hydrogels for the assessment of biofilm susceptibility towards biocide treatments. AB - Poloxamer F127 is a non-toxic, di-block copolymer of polyoxyethylene and polyoxypropylene. Aqueous solutions (30% w/v) show thermoreversible gelation, being liquid at temperatures < 15 degrees C and robust gels at temperatures > 15 degrees C. Chilled poloxamer (30% in tryptone soya broth) was mixed with an inoculum of Pseudomonas aeruginosa (10(4) cfu ml-1) and placed as 100 microliters drops onto separate glass cover-slips. These were placed into sealed Petri dishes containing moistened cotton wool and incubated at 35 degrees C. Viable counts could be performed on the poloxamer gels by transfer of the coverslips to diluents at < 15 degrees C. Growth curves in the gels and in liquid batch cultures were indistinguishable from one another with stationary phase cell densities, being approximately 5 x 10(10) cfu ml-1 in each at 16 h. SDS-PAGE of cell envelope preparations showed the poloxamer-grown cells to exhibit a biofilm rather than planktonic phenotype. Susceptibility towards various concentrations of chlorhexidine, iodine and hydrogen peroxide was assessed for 10 min at 35 degrees C for suspensions of broth-grown cells and for incubated poloxamer-gels (1 and 16 h). The gels were immersed in biocide, on their glass supports, before transfer to neutralizer at 10 degrees C where dissolution was complete within 5 min. Further serial dilutions and plate counts were made. While modest decreases in susceptibility towards all biocides were associated with incorporation of the inoculum with the gel (1 h incubation), substantial changes were noted after prolonged incubation and adaptation to a biofilm phenotype (16 h incubation). The gel populations mimic the localized high cell densities observed in biofilms and will also be subject to the same nutrient and chemical gradients as found within natural biofilms. Thermoreversible gelation enables complete recovery of the test inoculum without further trauma. They therefore provide an effective model for assessing biofilm susceptibility towards biocides and would be suitable for screening programmes. PMID- 9871319 TI - Development of a sterile cold-smoked fish model. AB - It is difficult to create a successful sterile cold-smoked fish model, particularly when very fresh fish are not readily available. This study tested a low-dose ionization technique (1.5 and 3.0 kGy) as a means of eliminating residual flora and ensuring sterility without altering sensory qualities. Ionized cold-smoked salmon displayed no bacterial contamination during an 11 week storage period and was not considered as spoiled by a sensory panel. In comparison, non ionized smoked salmon, although aseptically processed, contained a bacterial flora responsible for its spoilage. This model allows assessment of the spoilage potential or activity of isolated bacterial strains as well as independent studies of bacterial and non-bacterial reactions in cold-smoked fish. PMID- 9871320 TI - Characterization of lactococci isolated from milk produced in the Camembert region of Normandy. AB - Thirty-eight Lactococcus strains, isolated from raw milk produced in two dairy areas in Normandy, were identified at the phenotypic level. Only Lactococcus lactis strains with the lactis phenotype were found in the milk samples. Most strains fermented lactose (97%) and showed proteinase activity (76%). Isolates were characterized by RAPD technique and rRNA gene restriction analysis. More L. lactis strains with the lactis genotype were found in the first area, while L. lactis strains with the cremoris genotype predominated in the second area. RAPD was more efficient than rRNA gene restriction analysis in differentiating between strains with the subsp. lactis genotype. For L. lactis with the subsp. cremoris genotype, the second method gave a better result but there was poor discrimination between strains. Plasmid profiles were determined. Patterns ranged in size from 1.3 to 16.5 kbp, and 29 different profiles were found. Six groups of strains were determined, five of which were specific for the area of origin. It is suggested that the region of manufacture could influence organoleptic properties of cheeses because of different Lactococcus strains in the raw milk used for cheese making. PMID- 9871321 TI - Factors influencing a suspension test method for antimicrobial activity of disinfectants. AB - Factors influencing the numbers of Escherichia coli DSM 682 and Staphylococcus aureus ATCC 6538 surviving exposure to disinfectants were evaluated by factorial design. Aerobic conditions during pre-cultivation rendered E. coli more resistant to the lethal activity of benzalkonium chloride (BC) and a disinfectant containing grape fruit extract (GSE), whereas Staph. aureus became more sensitive. The degree of shaking and the pre-growth medium (tryptone soy broth or Mueller-Hinton broth) did not influence the result of the bactericidal test. The number of E. coli surviving BC treatment was significantly lower if the neutralizing broth contained thiosulphate, plate pouring was used instead of plate spreading, or the plates were incubated at 37 instead of 30 degrees C. The negative effect of plate pouring was also found with Staph. aureus. The use of filtration without prior neutralization of the disinfectant decreased the numbers of chlorine-treated, but not BC-treated, E. coli. The results showed that rigorous standardization is necessary to obtain good reproducibility of bactericidal suspension tests. PMID- 9871322 TI - Synergist effect of sucrose fatty acid esters on nisin inhibition of gram positive bacteria. AB - Nisin in combination with the sucrose fatty acid esters, sucrose palmitate (P 1570 and P-1670) or sucrose stearate (S-1570 and S-1670) was tested against a range of Gram-negative and Gram-positive bacteria. Initial liquid culture investigation showed that the sugar ester P-1670 resulted in a synergist enhancement of the bacteriostatic activity of nisin against Gram-positive bacteria and not Gram-negative bacteria. Some enhancement of the bactericidal activity of nisin against Listeria monocytogenes was also observed. This increased nisin inhibitory effect was confirmed on solid media using plates with gradients of pH and NaCl. Synergism was observed with all four sucrose fatty acid esters, which enhanced the antimicrobial activity of nisin against several strains of L. monocytogenes, Bacillus cereus (both cells and spores), Lactobacillus plantarum and Staphylococcus aureus. The combination of nisin and the sucrose fatty acid esters showed no inhibition of Gram-negative bacteria (Salmonella enteritidis, Salm. typhimurium and Pseudomonas aeruginosa). PMID- 9871323 TI - Factors affecting biosynthesis by Xanthomonas albilineans of albicidin antibiotics and phytotoxins. AB - Albicidins are important factors in systemic pathogenesis by Xanthomonas albilineans, which causes the devastating leaf scald disease of sugar cane. They are also of substantial interest as antibiotics that selectively block prokaryote DNA replication. Albicidin biosynthesis is highly sensitive to medium composition. An optimized, chemically defined medium (SMG3) yielded 30-fold more albicidin from half the accumulated biomass, relative to sucrose peptone (SP) medium. Phosphate starvation stimulated albicidin production in SMG3 and SP media. Addition of other amino acids, ammonium ions or peptones to the defined medium increased the growth rate of X. albilineans XA3, but differentially inhibited albicidin biosynthesis. Knowledge of these factors indicates new approaches to understanding mechanisms of pathogenesis and resistance to sugar cane leaf scald disease, and to strain improvement for production of albicidin antibiotics. PMID- 9871324 TI - Phospholipid molecular species distribution of Porphyromonas asaccharolytica ATCC 25260T: effects of temperature, culture age and pH. AB - The aim of this study was to determine the extent to which phospholipid molecular species profiles are affected by different environmental factors in Porphyromonas asaccharolytica ATCC 25260T. Phospholipids were analysed by Fast Atom Bombardment Mass Spectrometry (FAB-MS) in negative-ion mode. Under standard growth conditions (37 degrees C, pH 7.0, 48 h), the most intense high mass anions were m/z 653 and 662. The latter is consistent with the expected presence of PE (30:0). The only changes in profiles were quantitative. These were compared using the Pearson Coefficient of Linear Correlation. The r-values for initial pH comparisons ranged from 0.82 (pH 7.0 vs pH 6.0) to 1.00 (pH 5.0 vs pH 8.0), for incubation period, from 0.86 (48 vs 72 h) to 0.97 (96 vs 168 h), and for temperature, from 0.57 (40 vs 37 degrees C) to 0.96 (37 vs 36 degrees C). Differences were also seen when plates were incubated in anaerobe jars as opposed to an anaerobic work station (r = 0.75). It is concluded that it is essential to standardize growth parameters, and to use an anaerobe jar or an anaerobe work station, but not both. PMID- 9871325 TI - A possible role of ProP, ProU and CaiT in osmoprotection of Escherichia coli by carnitine. AB - Exogenously provided carnitine (beta-hydroxy-L-tau-N-trimethyl aminobutyrate) was found to stimulate aerobic growth of enterohaemorrhagic Escherichia coli O157:H7 in a medium of inhibitory osmotic strength. Its osmoprotective ability is comparable with that of betaine. As carnitine is an important compound in mammalian tissues, it is suggested that it might play a role in the growth of the pathogen on low water activity (aw) meat products. Using specific uptake mutants of E. coli K-12, it was established that, under osmotic stress, carnitine accumulates in the cytoplasm following import through the ProP and ProU transport systems. Betaine and carnitine also protect E. coli cells while growing anaerobically at inhibitory osmolarity. Under these conditions, an E. coli K-12 strain with lesions in both proP and proU accumulates low levels of L-carnitine but fails to accumulate betaine when these compounds are supplied in the external medium. This is probably a result of uptake of L-carnitine by the secondary transporter CaiT. The caiT gene forms part of the caiTABCDE operon which encodes the carnitine pathway, and is transcribed during anaerobic growth in the presence of carnitine. However, further experiments revealed that the carnitine pathway, including CaiT, does not play a significant role in osmoregulation of E. coli during anaerobiosis. Together, the results indicate that ProP and ProU are the sole transport systems involved in carnitine influx, both in aerobically and anaerobically osmotically stressed E. coli cells. PMID- 9871326 TI - Comparison of the inhibitory effect of sorbic acid and amphotericin B on Saccharomyces cerevisiae: is growth inhibition dependent on reduced intracellular pH? AB - The effects of sorbic acid and amphotericin B on the growth and intracellular pH (pHi) of Saccharomyces cerevisiae were studied and compared. Past evidence has suggested that the inhibitory action of sorbic acid on yeast is due to reduction of pHi per se. However, using a novel method to measure pHi in growing cells, little correlation was found between reduced growth rate on exposure to sorbic acid and reduction of pHi. In fact, growth inhibition correlated with an increase in the intracellular ADP/ATP ratio due to increased ATP consumption by the cells. This was partly attributed to the activation of protective mechanisms, such as increased proton pumping by the membrane H(+)-ATPase, which ensured that pHi did not decline when cells were exposed to sorbic acid. Therefore, the available evidence suggested that the inhibitory action of sorbic acid was due to the induction of an energetically expensive protective mechanism that compensated for any disruption of pHi homeostasis but resulted in less available energy for normal growth. In contrast to sorbic acid, with amphotericin B there was a direct correlation between growth inhibition and reduction of pHi due to the uncoupling effect of this compound on the plasma membrane. The inhibitory effect of amphotericin B was consistent with membrane disruption, or 'proton-uncoupling' leading to growth inhibition due to proton influx, decline in pHi and partial dissipation of the proton gradient. PMID- 9871327 TI - Note: characterization of Vibrio cholerae O139 Bengal isolated from water in Malaysia. AB - Four Vibrio cholerae O139 Bengal strains isolated from surface water were characterized by antibiotic resistance, plasmid profile, presence of cholera toxin gene and random amplification of polymorphic DNA (RAPD) analysis. All four strains exhibit multiple resistance towards the antibiotics tested with a multiple antibiotic resistance index of 0.5-0.66, and harboured a 2.0 MDa non conjugative plasmid. The Vibrio cholerae O139 Bengal were positive for the cholera toxin gene. Antibiotyping and random amplification of polymorphic DNA analysis with four primers proved to be useful in discriminating the isolates. RAPD proved to be more sensitive. These results reveal that there is significant genetic diversity among the Vibrio cholerae O139 Bengal strains studied. PMID- 9871328 TI - The occurrence of fungi along the Red Sea coast and variability among isolates of Fusarium as revealed by isozyme analysis. AB - Nineteen identified species which belong to nine fungal genera were recovered from 14 samples collected from different sites of the Red Sea governorate. The aquatic fungal genera were Allomyces, Dictyuchus, Saprolegnia and Pythium while, the terrestrial fungal genera were Aspergillus, Penicillium, Fusarium, Neurospora and Rhizopus. Aspergillus was the most frequent genus, represented by seven species, of which A. niger, A. flavus and A. ustus were the most common. Penicillium was of occurred less frequently and was represented by two species, while Fusarium was isolated unfrequently and contributed four species. The remaining genera were unfrequent or rare and were each represented by one species. In addition, two electrophoretic isozyme patterns, esterase and glutamate oxalate transaminase (GOT), were determined to measure variability among 10 isolates of Fusarium. The results revealed that the tested fungi differed from each other in one or more esterase bands, except that F. moniliforme isolated from Safaga and from 40 Kilometers south of El-Kaussier yielded similar banding pattern. The activity of GOT was observed in the samples of F. solani and F. oxysporum and not detected in other isolates of Fusarium. The results indicated that F. solani differed from F. oxysporum in the isozymes of GOT, while no differences were observed between the isolated of the same species. PMID- 9871329 TI - Search for bacteriophages spontaneously occurring in cultures of haemolytic intestinal spirochaetes of human and animal origin. AB - An electron microscopic survey of the occurrence of bacteriophages which appear spontaneously in cultures of haemolytic intestinal spirochaetes of human and animal origin was made. Excluding one isometric tailed phage particle which was observed in the form of free particle in proximity to a spirochaete of the w beta HIS strain HRM18, bacteriophages were never observed while examining cells of 21 weakly beta-haemolytic human intestinal spirochaetes (w beta HIS), swine Serpulina pilosicoli strain P43/6/78, and the avian strain 1380, although 50-100 cells of each spirochaetal strain were analysed. Isometric tailed bacteriophages were found associated with only three out of the 100 cells of strongly beta haemolytic swine Serpulina hyodysenteriae strain P18A comparatively analysed. According to our results and previous published reports, the occurrence of bacteriophages which appear spontaneously in cultures of intestinal spirochaetes is a rare event. PMID- 9871330 TI - Bacteriophages induced from weakly beta-haemolytic human intestinal spirochaetes by mitomycin C. AB - A comparative electron microscopic analysis of weakly beta-haemolytic spirochaetes related to human and animal intestinal spirochaetosis was done in order to search for the presence of inducible bacteriophages associated with these spirochaetes. Bacteriophages were detected at the electron microscope after experimental induction with mitomycin C in 4 strains of weakly beta-haemolytic spirochaetes related to human intestinal spirochaetosis, in Serpulina pilosicoli strain P43/6/78, the causative agent of swine intestinal spirochaetosis, in a spirochaetal strain related to avian intestinal spirochaetosis, and in Serpulina hyodysenteriae, strain P18A, the causative agent of swine dysentery, which was comparatively analysed as control. All phage-particles observed in both human and animal intestinal spirochaetes were morphologically similar with an isometric head of 45 nm diameter and a tail 63-70 nm long and 7-12 nm width. The presence of morphologically similar phages in all the haemolytic intestinal spirochaetes of human and animal origin analysed in this study opens some important questions, about the genetic relationship of phages present in pathogenic intestinal spirochaetes, their host range, and the possibility of natural gene transfer among pathogenic haemolytic intestinal spirochaetes of human and animal origin. PMID- 9871331 TI - Pathogenic action of Penicillium species on mosquito vectors of human tropical diseases. AB - In vitro bioassays were performed in order to assess the pathogenicity of 13 Penicillium strains in 2nd stage larvae of Aedes aegypti, Aedes fluviatilis, Anopheles aquasalis and Culex quinquefasciatus. Mortality rates began in the first 24 hours, ranging from 0 to 100%. P. corylophilum, P. fellutanum, P. implicatum, P. janthinellum, P. viridicatum and P. waksmanii species tested on Aedes aegypti larvae and at different concentrations presented mortality rates from 0 to 6.6%. These species, when tested on Aedes fluviatilis, Anopheles aquasalis and Culex quinquefasciatus larvae, presented a mortality rate of 3.33% to 100%. Susceptibility of Aedes fluviatilis, Aedes aegypti, Anopheles aquasalis and Culex quinquefasciatus mosquitoes to the tested agents, turn P. corylophilum and P. janthinellum into candidates for potential use in biological control of vectors. PMID- 9871332 TI - Penicillium species in mosquitoes from two Brazilian regions. AB - Penicillium species were isolated from 1073 adult mosquitoes and larvae of Aedes spp., Anopheles spp., Culex spp. and Mansonia spp. captured in the northern and southeastern regions of Brazil. In a total of 24 collections, 198 Penicillium strains were identified in 13 species: Penicillium canescens, P. chrysogenum, P. citrinum, P. corylophilum, P. decumbens, P. expansum, P. fellutanum, P. implicatum, P. janthinellum, P. oxalicum, P. purpurogenum, P. viridicatum and P. waksmanii. The total isolation frequency of the Penicillium species in the investigated mosquitoes was: Anopheles spp. (51.5%), Aedes spp. (15.1%), Culex spp. (23.7%) and Mansonia spp. (10.1%). The Penicillium species with the highest incidence in the mosquito genera were: P. corylophilum, P. janthinellum, P. fellutanum and P. waksmanii. The highest number of mosquitoes were captured in the state of Rio de Janeiro, followed by Rondonia and Minas Gerais. PMID- 9871333 TI - Degradation of 2-chlorophenol and formation of 2-chloro-1,4-benzoquinone by mycelia and cell-free crude culture liquids of Trametes versicolor in relation to extracellular laccase activity. AB - The white-rot fungus Trametes versicolor was used to study the influence of extracellular laccase activity on the degradation of 2-chlorophenol (2-CP) and the formation of metabolites under conditions, characterized by the absence of other phenol-oxidizing enzymes. 2-CP enhanced the production of extracellular laccase by fungal mycelia. The formation of the metabolite 2-chloro-1,4 benzoquinone (2-CIBQ) was found to be correlated with extracellular laccase activity. In cell-free crude culture liquids laccase was responsible for the oxidation of 2-CP. In contrast to this, the disappearance of 2-CP caused by the entire organism did not correlate with extracellular laccase activity. The primary oxidative step during the degradation of this compound can thus only partially be attributed to extracellular laccase; indicating the involvement of cell-bound processes. PMID- 9871334 TI - Serum sensitivity and cell surface hydrophobicity of Klebsiella pneumoniae treated with gentamicin, tobramycin and amikacin. AB - A suppression of bacterial growth of Klebsiella pneumoniae after 30 min treatment with gentamicin, tobramycin and amikacin at suprainhibitory concentrations was found (postantibiotic effect, PAE). The antibiotics at a concentration of 2 x MIC induced PAEs in the range of 0.5-1.7 h, PAEs manifested by aminoglycosides at a concentration of 4 x MIC varied between 0.8-3.3 h. Susceptibility of exposed bacteria to serum bactericidal activity was efficiently enhanced compared with controls (without antibiotics). A survival of treated bacteria was between 12.7 36.6% (2 x MIC) or 5.0-8.3% (4 x MIC). The percentage of viable nontreated bacteria moved between 70.2-83.0% at these conditions. Surface hydrophobicity of bacteria exposed to aminoglycosides was only slightly increased. The results indicate that exposure of K. pneumoniae to a suprainhibitory concentrations of aminoglycosides in vitro enhanced the susceptibility of this strain to human serum bactericidal activity. Whether this phenomenon occurs in vivo, remains to be determined. PMID- 9871335 TI - The gluconate high affinity transport of GntI in Escherichia coli involves a multicomponent complex system. AB - Within the main system for gluconate utilization in E. coli, the gntT gene (located at the minute 76.4) that encodes a permease, is currently the only element involved in the high affinity transport. In this paper, the nucleotide sequence of the upstream region of this locus was determined. Two open reading frames of 729 bp (gntX) and 573 bp (gntY) were identified as additional gnt genes by complementation studies. Our observations suggest that these loci might conform an operon distinct of gntT under the control of the gntR gene product. Such operon encodes a gluconate periplasmic binding protein (GntX) and a putative membrane-bound protein (GntY). These products and the permease encoded by the gntT gene seem to conform a high-affinity complex transport system for gluconate. We suggest that this novel system could belong to the TRAP transporters. PMID- 9871336 TI - N'-methylniphimycin, a novel minor congener of niphimycin from Streptomyces spec. 57-13. AB - A novel natural niphimycin analog, N'-methylniphimycin was isolated from the culture broth of the Streptomyces spec. 57-13. The chemical constitution was elucidated from the physico-chemical properties, NMR techniques and mass spectrometry to be a 36-membered macrolide related to azalomycin F5a, shurimycin B and RS-22C. N'-methylniphimycin displayed moderate activity against some yeasts and filamentous fungi. PMID- 9871337 TI - Examination of micro-tip reversed-phase liquid chromatographic extraction of peptide pools for mass spectrometric analysis. AB - Mass spectrometry occupies a central position in most current protein identification schemes. So-called 'mass fingerprinting' techniques rely on composite mass patterns of proteolytic fragments, or dissociation products thereof, to query databases. Keys to successful analysis of ever smaller amounts are sensitivity and complete spectral information, both of which depend for a large part on proper sample preparation. Clean-up and concentration of peptide mixtures over eppendorf gel loading tips filled with chromatographic media (i.e. 'micro-tips') are believed to be quite useful in this regard. We have studied quantitative and qualitative aspects of polypeptide extraction using these small manual devices. Optimization of sample volume and additives, micro-tip bed volume, and eluent composition and volume, all contribute to effective recovery (approximately 65-70%, on average). Improper digest conditions can, in fact, lead to far bigger losses, suggesting the need for at least trace amounts of Zwittergent 3-16. Of particular interest is our finding that partial fractionation, obtained by two-step micro-tip elution, generally results in more and better signals during subsequent mass analysis. Thus, by using optimized micro-tips, in combination with adequate sample handling and instrumentation, direct mass spectrometric identification can be routinely and successfully done in any resource facility type setting. PMID- 9871338 TI - Column liquid chromatography determination of vitamins A and E in powdered milk and local flour: a validation procedure. AB - A high-performance liquid chromatography method was developed for the simultaneous routine determination of vitamins A and E in powdered milk and flour made from local plants and purchased from open markets in the Ivory Coast. The method involves saponification followed by extraction with a mixture of organic solvents. The vitamins were resolved by reversed-phase HPLC and detection at a single wavelength. The main tests of method validation were applied to the procedure. The results show the reliability of the analytical method for the intended application. PMID- 9871339 TI - Determination of benzene, toluene, ethylbenzene and xylenes in indoor air at environmental levels using diffusive samplers in combination with headspace solid phase microextraction and high-resolution gas chromatography-flame ionization detection. AB - An improved analytical method for passive air sampling is presented based on a combination of commercially available diffusive samplers with headspace solid phase microextraction and high-resolution gas chromatography with flame ionization detection (HRGC-FID). This procedure is targeted for short-term BTEX (benzene, toluene, ethylbenzene and o-, m- and p-xylenes) determinations at environmental concentrations and can be applied for sampling intervals between 30 min and 24 h. The analytes are adsorbed onto the charcoal pad of a passive sampler and then extracted with carbon disulphide-methanol. After removal of the carbon disulphide by xanthation, the BTEXs are enriched on a Carboxen SPME fiber, thermally desorbed and analysed by HRGC-FID. Detection limits for a sampling interval of 2 h are between 0.4 and 2 micrograms/m3, within-series precision ranges between 6.6 and 12.8%, day-to-day precision is between 11.1 and 15.2%. The results obtained with this procedure are validated by comparison with active sampling. Detection limits and a further reduction of the sampling time are limited by blanks of the chemicals and the diffusive samplers. Procedures to eliminate these blanks are described in detail. Applications such as the determination of BTEXs in indoor air inside buildings, inside a train and a car are presented, indicating the usefulness of the described procedure for short term measurements of environmental BTEX concentrations. An advantage of passive samplers is the storage stability for at least six months, which is essential for its use in large epidemiological studies. PMID- 9871340 TI - Liquid chromatographic method for determining added folic acid in fortified cereal products. AB - Reversed-phase ion-pair high-performance liquid chromatography (HPLC) was coupled with detection by UV absorption (280 nm) for separation and quantitation of added folic acid (FA) in fortified cereal based foods. A simple and rapid liquid-solid extraction method, combined with enzymatic digestion, to recover FA from the sample matrices is also presented. The quantitation of added FA was achieved in products including corn (maize), wheat-, rice- and oat-based cereal breakfast foods fortified at 25% and 100% of the reference daily intake (RDI). The retention time for FA was ca. 15 min, and the detection limit was 2 ng/20 microliters injection for standard FA. When FA was added to unfortified samples of wheat flour at concentrations of 3.08 or 20.0 micrograms/g, recoveries were 93% and 96%, respectively. Comparison of HPLC results with those of a standard microbiological assay has shown quite good agreement (r = 0.998). A solid-phase extraction clean-up procedure has also been developed for use with samples fortified with low levels of FA, where interferences may otherwise hinder quantitation. PMID- 9871341 TI - Effects of hypothermic hypoxia on anaerobic energy metabolism in isolated anuran livers. AB - Many lower vertebrates (reptilian and amphibian species) are capable of surviving natural episodes of hypoxia and hypothermia. It is by specific metabolic adaptations that anurans are able to tolerate prolonged exposure to harsh environmental stresses. In this study, it was hypothesized that livers from an aquatic frog would possess an inherent metabolic ability to sustain high levels of ATP in an isolated organ system, providing insight into a metabolic system that is well-adapted for low temperature in vitro organ storage. Frogs of the species, R. pipiens were acclimated at 20 degrees C and at 5 degrees C. Livers were preserved using a clinical preservation solution after flushing. Livers from 20 degrees C-acclimated frogs were stored at 20 degrees C and 5 degrees C and livers from 5 degrees C-acclimated frogs were stored at 5 degrees C. The results indicated that hepatic adenylate status was maintained for 96 h during 5 degrees C storage, but not longer than 4-10 h during 20 degrees C storage. In livers from 5 degrees C-acclimated animals subjected to 5 degrees C storage, ATP was maintained at 100% throughout the 96-h period. Warm acclimation (20 degrees C) and 20 degrees C storage resulted in poorer maintenance of ATP; energy charge values dropped to 0.50 within 2 h and by 24 h, only 24% of control ATP remained. Lactate levels remained less than 25 mumol/g dry weight in all 5 degrees C-stored livers; 20 degrees C-stored livers exhibited greater accumulation of this anaerobic endproduct (lactate reached 45-50 mumol/g by 10 h). The data imply that hepatic adenylate status is largely dependent on exposure to hypothermic hypoxia and although small amounts of ATP were accounted for by anaerobic glycolysis, there must have been either a substantial reduction in cellular energy utilization or an efficient use of low oxygen tensions. PMID- 9871342 TI - Polyunsaturated fatty acids in serum lipids of reindeer during the close postnatal period. AB - We examined serum fatty acid composition in reindeer during the close postnatal period (from < 8 h to 3 weeks) by using maternal serum as a reference point and focusing on the proportions of polyunsaturated fatty acids (PUFAs) in serum lipids. A striking dissimilarity was found in the serum PUFAs between the neonatals and their mothers. In particular, the proportions of linoleic acid (18:2) and alpha-linolenic acid in serum cholesteryl esters and phospholipids of the newborn reindeer were significantly lower than those of the mothers. Furthermore, serum phospholipids of the newborns had lower arachidonic acid and docosapentaenoic acid but higher docosahexaenoic acid proportions than the maternal phospholipids. Although the proportions of the principal C18 PUFAs were low in reindeer milk, they increased sharply in serum cholesteryl esters and phospholipids of the calves during the first few days after birth. In particular, there were significant positive correlations in the proportions of 18:2 between serum and milk lipids. We conclude that the proportions of the serum C18 PUFAs are low in the newborn reindeer, but they are increased during the close perinatal period by a rate which suggests an efficient mechanism for selective retention of these fatty acids from milk lipids. PMID- 9871343 TI - Copper exposure impairs intra- and extracellular acid-base regulation during hypercapnia in the fresh water rainbow trout (Oncorhynchus mykiss). AB - In order to evaluate the impact of water-borne copper on acid-base regulation in fresh water rainbow trout, chronically cannulated fish were exposed to copper (0.6 mg 1(-1)), hypercapnia (water PCO2 of 6 mmHg) or a combination of copper and hypercapnia, while a fourth untreated group served as the control. Blood samples obtained at 0 h, 4 h and 24 h were analysed for acid-base status, ion concentrations and respiratory parameters. Tissue samples from caudal skeletal muscle, liver and gill filaments were examined for intracellular acid-base status, ion- and water contents, and copper concentration. Exposure to copper alone elicited a small extracellular metabolic alkalosis, no changes in arterial PO2, and a minor decrease in plasma ion concentrations. Hypercapnia alone increased arterial PCO2 from approximately 2 mmHg to 7.2 mmHg, but the extracellular respiratory acidosis present at 4 h was almost completely compensated at 24 h due to an increase in plasma bicarbonate concentration [HCO3 ] from 8.1 mM to 24.4 mM. Combined exposure to hypercapnia and copper resulted in a slightly larger acidosis at 4 h, and the fish failed to restore extracellular pH at 24 h, because plasma [HCO3-] only increased to 16.3 mM. Fish exposed to hypercapnia and copper also showed a delayed recovery of intracellular pH in skeletal muscle, compared to fish exposure to hypercapnia only. Thus, copper exposure impaired both extracellular and intracellular acid-base regulation during hypercapnia. When seen in connection with only minor effects of copper on osmoregulatory and respiratory parameters, the reduced ability to regulate acid base suggests that acid-base regulation may be one of the most copper-sensitive branchial functions. PMID- 9871344 TI - Opposite effects of cooling on twitch contractions of skeletal muscle isolated from tropical toads (Leptodactylidae) and northern frogs (Ranidae). AB - Cooling increases the twitch force of frog skeletal muscle (Rana temporaria; Rana pipiens), but decreases the twitch force of tropical toad muscle (Leptodactylus insularis). Action potentials and intramembranous charge movement in frog and toad fibers were slowed identically by cooling. Cooling increased the integral of twitch Ca2+ detected by aequorin in frog fibers (1.4-fold), while also decreasing the peak and slowing the rate of decay. Conversely, cooling decreased the integral (0.6-fold) and the peak of twitch Ca2+ in toad fibers, without affecting the rate of decay. The difference in entire Ca2+ transients may account for cold induced twitch potentiation in frogs and twitch paralysis in toads. In sustained contractions of toad fibers, cooling markedly decreased maximum force caused by: (i) tetanic stimulation, (ii) two-microelectrode voltage clamp steps, (iii) high [K+], or (iv) caffeine. Maximum force in sustained contractions was decreased moderately by cooling frog fibers. Rapid rewarming and simultaneous removal of high [K+] or caffeine during a sustained contraction, caused toad muscle force to rise towards the value corresponding to the warm temperature. This did not occur after removing high [K+] or caffeine from toad fibers kept in the cold. Transmission electron micrographs showed no relevant structural differences. Parvalbumins are thought to promote relaxation of frog muscle in the cold. The unique parvalbumin isoforms in toad muscle apparently lack this property. PMID- 9871345 TI - Administration of a 5HT3 receptor antagonist increases the intake of diets containing Eucalyptus secondary metabolites by marsupials. AB - The effect of a naturally occurring plant phenolic constituent (the acylphloroglucinol derivative, jensenone, derived from Eucalyptus jensenii) on the food intake of two folivorous marsupials, the common ringtail (Pseudocheirus peregrinus) and the common brushtail possum (Trichosurus vulpecula) was studied. When fed diets containing varying concentrations of jensenone, both species regulated their intake of jensenone so as not to exceed a ceiling intake. This ceiling was about twice as high for common ringtails as for common brushtails from northern Australia. Southern populations of common ringtails showed greatly reduced capacities to tolerate jensenone. When common brushtails were injected (0.5 mg.kg-0.75 body mass) with ondansetron (a selective antagonist of serotonin 5HT3 receptors), they ate significantly more jensenone than animals injected with physiological saline. The same pattern was observed when common ringtails were fed diets containing both jensenone and ondansetron (0.0035 mg.g-1 wet mass of diet). Ondansetron injection had no effect on food intake when the food did not contain jensenone while the addition of higher doses of ondansetron to diets of common ringtails very slightly reduced food intakes of a non-jensenone diet. When common brushtails were given 50 mg of jensenone by gastric lavage, their average subsequent intake of dietary jensenone matched the difference between the daily threshold and the dose given, although the response of individuals was highly variable. Lavage with water alone had no effect on subsequent jensenone intake compared with the pre-dose period. We interpret these results as evidence that the antifeedant effects of jensenone and related compounds are partly mediated by serotonin action on 5HT3 receptors most likely via "nausea" to condition a food aversion. PMID- 9871346 TI - Rhythmicity of torpor in a marsupial hibernator, the mountain pygmy-possum (Burramys parvus), under natural and laboratory conditions. AB - Circadian rhythms have been observed in most mammals, but their importance and function remain controversial with respect to daily cycles during hibernation. We investigated the timing of arousals from and entries into hibernation for both free-living and captive mountain pygmy-possums (Burramys parvus). Under both natural and laboratory conditions most arousals and entries were entrained with the light-dark cycle. Entries occurred mainly during the night and arousals preferably around dusk, which coincides with the onset of the normal activity phase for the nocturnal pygmy-possums. This entrainment prevailed throughout the hibernation season although only the laboratory animals were constantly subjected to photoperiodic stimuli, whereas under natural conditions hibernacula are shielded from photic cues and diurnal temperature fluctuations. Nevertheless, possums left their hibernacula frequently throughout winter and were occasionally trapped close to the snow surface suggesting that during the periods of post arousal normothermia they can be exposed to environmental stimuli. It thus appears that the synchronisation with the photocycle was governed by a temperature-compensated circadian clock which was reset periodically during short activity periods. For the mountain pygmy-possum, entrainment with the photocycle probably has two functions: 1. Entrainment ensures that foraging bouts during the hibernation season remain synchronised with the dark phase. 2. Information about the prevailing climatic conditions sampled during short activity periods enables them to time final spring emergence from hibernation when snow melt begins and ensures that the breeding season can commence as early as possible. PMID- 9871347 TI - Redox potential affects the measured heat resistance of Escherichia coli O157:H7 independently of oxygen concentration. AB - Cells of Escherichia coli O157:H7 were heat-treated at 59 degrees C and enumerated in (i) anaerobic medium with a low redox potential, (ii) anaerobic media with the oxidizing agents potassium ferricyanide or 2,6-dichloroindophenol (DPIP) added to raise the redox potential, (iii) aerobic medium with a high redox potential and (iv) aerobic medium with the reducing agent dithiothreitol added to lower the redox potential. The measured heat-resistance was greatest when the enumeration medium was highly anaerobic due to the absence of oxygen and the presence of hydrogen and cysteine HCl. Measured heat resistance was influenced by the redox potential of the enumeration medium independently of the chemical used to adjust it and therefore, independently of the presence of oxygen. Sub-lethally heat-damaged cells regained their ability to grow in media of high redox potential at a similar rate whether the redox potential was increased by the addition of potassium ferricyanide, DPIP or oxygen. PMID- 9871348 TI - Recovery of Cryptosporidium oocysts from small and large volume water samples using a compressed foam filter system. AB - A novel filter system comprising open cell reticulated foam rings compressed between retaining plates and fitted into a filtration housing was evaluated for the recovery of oocysts of Cryptosporidium from water. Mean recoveries of 90.2% from seeded small and large volume (100-2000 l) tap water samples, and 88.8% from 10-20 l river water samples, were achieved. Following a simple potassium citrate flotation concentrate clean-up procedure, mean recoveries were 56.7% for the tap water samples and 60.9% for river water samples. This represents a marked improvement in capture and recovery of Cryptosporidium oocysts from water compared with conventional polypropylene wound cartridge filters and membrane filters. PMID- 9871349 TI - Evaluation of two media for the membrane filtration enumeration of Clostridium perfringens from water. AB - Two media (mCP medium and Tryptose Sulphite Cycloserine (TSC) agar) were evaluated for recovery of Clostridium perfringens in environmental and part treated drinking water. For laboratory strains of Clostridium, mCP was more selective and specific for Cl. perfringens than TSC, but was markedly less efficient for the enumeration of both vegetative cells and spores. For samples of river water and part-treated drinking water, TSC recovered significantly greater numbers of Cl. perfringens than mCP. In contrast to previous reports, there was a significant number of false presumptive positive and negative isolates on mCP. TSC is a more suitable medium for the routine monitoring of water supplies for the presence of Cl. perfringens. PMID- 9871350 TI - Bactericidal activity of catechin-copper (II) complexes on Escherichia coli ATCC11775 in the absence of hydrogen peroxide. AB - Washed Escherichia coli ATCC11775 cells were killed by (-)-epigallocatechin (EGC) in the presence of a non-lethal concentration of Cu2+ (1 mumol l-1) without additional H2O2, but not by (-)-epicatechin (EC). EGC alone (< 0.1 mmol l-1) did not reduce the viability of the cells. The survival curve obtained in the presence of EGC and Cu2+ was similar to that obtained in the presence of (-) adrenaline (EN) and Cu2+. PMID- 9871351 TI - Aerobic growth and survival of Campylobacter jejuni in food and stream water. AB - When 40 Campylobacter jejuni isolates from human clinical cases, raw chicken and water were tested, 29 (72.5%) could be adapted to grow on nutrient agar under aerobic conditions. Once adapted, these isolates could grow on repeated aerobic subculture. An aerobically-grown Camp. jejuni isolate survived almost as well as the same isolate grown microaerophilically in sterile chicken mince at 5 degrees C, and survival of a cocktail of Camp. jejuni isolates under both atmospheres was comparable at 25 degrees C. However, at 37 degrees C, the decline in numbers of the aerobically-grown cells was greater. Survival of cells on chicken nuggets was poorer than in chicken mince. In filter-sterilized stream water incubated aerobically at 5 degrees C, survival of inocula grown under different atmospheres was again similar, but slightly better with the microaerophically-grown cells. Adaptation to aerobic growth was not found to enhance survival under aerobic conditions. PMID- 9871352 TI - Synergistic antimicrobial effect of nisin whey permeate and lactic acid on microbes isolated from fish. AB - The antimicrobial activity of a combination of lactic acid and whey permeate fermented by a nisin-producing Lactococcus lactis strain was tested by the agar diffusion method using bacteria isolated from fish as test organisms. Lactic acid inhibited all bacterial strains studied, but nisin whey permeate inhibited Gram positive bacteria only. The combination was more effective than lactic acid alone against Pseudomonas fluorescens and Staphylococcus hominis isolated from fish, and Pseudomonas aeruginosa ATCC9721 and Micrococcus luteus ATCC9341. PMID- 9871353 TI - Isolation of Aeromonas salmonicida in association with purple-pigmented bacteria in sediment from a Scottish loch. AB - Aeromonas salmonicida was recovered in close association with an unidentified purple-pigmented organism, which was isolated from sediment in a Scottish loch during November (1997) and February (1998). However, there has not been any evidence of A. salmonicida infections, specifically furunculosis, associated with the fish in this loch. PMID- 9871354 TI - Evaluation of fluorochromes for flow cytometric detection of Cryptosporidium parvum oocysts labelled by fluorescent in situ hybridization. AB - Oligonucleotide probes specific to Cryptosporidium parvum (CRY1) were conjugated with a range of fluorochromes. The fluorescence after in situ hybridization (FISH) labelling of oocysts and controls was assessed. The objective was to determine the most suitable conjugate for FISH labelling, followed by analysis with a 488 nm laser flow cytometer. The most promising candidate was fluorescein isothiocyanate but only when linked to the CRY1 probe via an 18-carbon spacer arm consisting of six ethylene glycol moieties. The use of the spacer increased fluorescent signals fivefold compared with an equivalent probe in which the FITC was linked directly to the 5'-amino group of the DNA. PMID- 9871355 TI - Natural milk fatty acids affect survival and invasiveness of Listeria monocytogenes. AB - The effects of 12 fatty acids, naturally occurring in milk from several mammalian species, on the survival and invasion ability of Listeria monocytogenes, a food borne pathogen, were determined. The survival was tested in the presence of 200 micrograms ml-1 fatty acids suspended in brain hearth infusion broth or in storage conditioning solution (NaCl 1%) of Mozzarella cheese, an Italian soft unripened cheese, at pH 7.0 or 5.0. Lauric (C12:0), linoleic (C18:2) and linolenic (C18:3) acids exerted the strongest bactericidal activity. The invasive efficiency of L. monocytogenes, determined in the Caco-2 enterocyte-like cell line, was strongly decreased in the presence of the fatty acids tested (from about 20 to 500-fold). This research suggests that naturally occurring fatty acids of milk, supplemented in milk derivatives, could affect both bacterial growth and invasiveness and consequently, could serve as barriers towards L. monocytogenes infection. PMID- 9871356 TI - PCR differentiation of Escherichia coli from other gram-negative bacteria using primers derived from the nucleotide sequences flanking the gene encoding the universal stress protein. AB - A PCR-based test was developed to differentiate Escherichia coli from other Gram negative bacteria. The assay employed primers derived from the nucleotide sequences flanking the gene encoding the universal stress protein (uspA) and was found to be highly specific for E. coli. An 884 base pair (bp) specific product was amplified from all of the E. coli tested (n = 45), whereas no amplification was associated with non-E. coli Gram-negative bacteria (n = 11). PMID- 9871357 TI - Effect of oscillatory high hydrostatic pressure treatments on Byssochlamys nivea ascospores suspended in fruit juice concentrates. AB - The effect of continuous (689 MPa with holding times of 5, 15 or 25 min) and oscillatory (one, three or five cycles at 689 MPa with holding times of 1 s) high hydrostatic pressure treatments on the viability of Byssochlamys nivea ascospores suspended in apple and cranberry juice concentrates adjusted by dilution to water activities (aw) of 0.98 and 0.94 was evaluated at 21 and 60 degrees C. Inactivation of the initial spore inocula was achieved after three or five cycles of oscillatory pressurization at 60 degrees C when the aw was 0.98 in both fruit juices. With aw 0.94, the initial inocula were reduced by less than 1 log-cycle after five pressure cycles. Inactivation was not observed within 25 min with continuous pressurization at 60 degrees C. In treatments at 21 degrees C, no effect on spore viability was observed with continuous or oscillatory treatments. PMID- 9871358 TI - Isolation of Salmonella resistant to hypochlorous acid from a poultry abattoir. AB - Salmonella is a Gram-negative bacterium that has been implicated in several food borne poisoning outbreaks. Poultry products are considered to be the major vehicles of food-borne poisoning caused by Salmonella. The resistance of Salmonella, isolated from different stages in a local poultry abattoir, to hypochlorous acid (HOCl) was studied. Isolates cultured in quarter strength Tryptone Soya Broth were treated with 72 ppm HOCl, incubated at 30 degrees C with shaking, and absorbance was measured at 660 nm every 20 min. A number of resistant isolates, which carried on growing following HOCl addition, were obtained. All the isolates from the bleeding stage were sensitive to HOCl, whereas those from the scalding stage were resistant to HOCl. Other stages had a population of resistant and sensitive isolates. It is evident that some Salmonella will grow in the presence of concentrations of HOCl deemed to be inhibitory. Hence, an effective concentration of HOCl must be applied in poultry abattoirs to ensure that all the Salmonella are eradicated. PMID- 9871359 TI - Of bourne, badgers and bovine TB. PMID- 9871360 TI - Rapid and definitive detection of Salmonella in foods by PCR. PMID- 9871361 TI - Reduction of caecal Listeria monocytogenes in Leghorn chicks following treatment with a competitive exclusion culture (PREEMPT) PMID- 9871363 TI - Characterization of a maize Ca(2+)-dependent protein kinase phosphorylating phosphoenolpyruvate carboxylase. AB - Phosphoenolpyruvate carboxylase (PEPC) [EC 4.1.1.31] of plants undergoes regulatory phosphorylation in response to light or nutritional conditions. However, the nature of protein kinase(s) for this phosphorylation has not yet been fully elucidated. We separated a Ca(2+)-requiring protein kinase from Ca(2+) independent one, both of which can phosphorylate maize leaf PEPC and characterized the former kinase after partial purification. Several lines of evidence indicated that the kinase is one of the characteristic Ca(2+)-dependent but calmodulin-independent protein kinase (CDPK). Although the M(r) of native CDPK was estimated to be about 100 kDa by gel permeation chromatography, in situ phosphorylation assay of CDPK in a SDS-polyacrylamide gel revealed that the subunit has an M(r) of about 50 kDa suggesting dimer formation or association with other protein(s). Several kinetic parameters were also obtained using PEPC as a substrate. Although the CDPK showed an ability of regulatory phosphorylation (Ser-15 in maize PEPC), no significant desensitization to feedback inhibitor, malate, could be observed presumably due to low extent of phosphorylation. The kinase was not specific to PEPC but phosphorylated a variety of synthetic peptides. The possible physiological role of this kinase was discussed. PMID- 9871362 TI - Molecular cloning and expression of the early auxin-responsive Aux/IAA gene family in Nicotiana tabacum. AB - Early auxin-regulated tobacco cDNAs, belonging to the Aux/IAA gene family have been isolated by screening of a cDNA library prepared from auxin-treated suspension-grown etiolated seedlings of Nicotiana tabacum. The probes used were either RT-PCR fragments or an insert resulting from mRNA differential display selection. All of them possessed the structural features which characterize the Aux/IAA gene products. The auxin response of three distinct Nt-iaa subclasses has been characterized in terms of kinetics, dose-response and specificity as several plant hormones and chemicals have been tested for their ability to alter Nt-iaa mRNA accumulation. Differences of auxin responses have been observed between the Nt-iaa analysed, revealing significant differences of regulation. The effect of the protein synthesis inhibitor cycloheximide suggested that Nt-iaa2.3, Nt-iaa4.3 and strictly related genes can be classified as primary auxin-responsive genes and Nt-iaa28 as a late one. The steady-state mRNA level of these Nt-iaa has also been determined in organs of tobacco plants. PMID- 9871364 TI - Structural studies of the vacuolar H(+)-pyrophosphatase: sequence analysis and identification of the residues modified by fluorescent cyclohexylcarbodiimide and maleimide. AB - We determined the amino acid residues of the H(+)-translocating inorganic pyrophosphatase (H(+)-PPase) of pumpkin which are covalently labeled by two fluorescent labeling reagents; N-cyclohexyl-N'-[4-(dimethyl amino)-alpha naphthyl] carbodiimide (NCD) and N-pyrenylmaleimide (NPM). NCD and NPM are fluorescent analogues of N,N-dicycrohexylcarbodiimide and N-ethylmaleimide, respectively, and inactivate H(+)-PPase activity. Excess Mg2+ protected the H(+) PPase from the inactivation by these reagents. Furthermore, we identified the cDNA clone encoding the pumpkin H(+)-PPase in order to determine the position of labeled residues. The nucleotide sequence of the cDNA clone contains a 2,304 bp open reading frame encoding a polypeptide with 768 amino acids. Chemical sequence analysis of fluorescent peptide fragments revealed that Glu749 located in the C terminal putative transmembrane alpha-helix was a NCD-labeled residue, and Cys632 was a NPM-labeled residue located in a putative cytosolic domain. The amino acid sequence of the region that includes Glu749 is highly conserved in H(+)-PPases from other plants and it also shows some sequence similarity with the region of the carbodiimide-reactive Glu (or Asp) of F0F1-ATPase c-subunit. The reactive glutamic acids in these proteins are located at the last C-terminal transmembrane alpha-helix. We also found that the H(+)-PPase shows significant amino acid sequence similarity to Kdp-ATPase A chain of E. coli. This similarity between the two different proteins suggest that they have evolved from a common ancestor and may utilize a common basic mechanism for ion transport. PMID- 9871365 TI - Characterization and expression of a new class of zinc finger protein that binds to silencer region of ascorbate oxidase gene. AB - A unique A/T-rich sequence (5'-AAAAAGTAAAAA-GTAAAAAAGTAAAAAG-3), referred to as the AGTA repeat, is found in the silencer region of the pumpkin ascorbate oxidase gene. A cDNA for protein (AOBP) that binds to the AGTA repeat was isolated from pumpkin by the southwestern method. The AOBP protein has a new class of zinc/DNA binding domain named Dof/MOA domain that is highly conserved in many plant proteins and is significantly related to those of steroid hormone receptors and GATA1. Gel retardation analysis indicated that AOBP bound to the AGTA repeat through the Dof/MOA domain. Metal chelators, 1,10-phenanthroline and EDTA, specifically inhibited the DNA binding of AOBP, indicating that metal coordination plays an important role in DNA binding of AOBP. Thus, the Dof/MOA domain acts as a zinc/DNA-binding domain in AOBP. Gel retardation analysis with mutated oligonucleotides suggested that the Dof/MOA domain recognized the AGTA core sequence. AOBP mRNA was expressed in mature tissues of pumpkin, but was expressed only in small amounts or was not expressed in growing tissues. Furthermore, the expression was auxin-independent. The expression pattern of AOBP and that of ascorbate oxidase did not show a positive correlation. PMID- 9871366 TI - Biological significance of divalent metal ion binding to 14-3-3 proteins in relationship to nitrate reductase inactivation. AB - In this report we address two questions regarding the regulation of phosphorylated nitrate reductase (pNR; EC 1.6.6.1) by 14-3-3 proteins. The first concerns the requirement for millimolar concentrations of a divalent cation in order to form the inactive pNR:14-3-3 complex at pH 7.5. The second concerns the reduced requirement for divalent cations at pH 6.5. In answering these questions we highlight a possible general mechanism involved in the regulation of 14-3-3 binding to target proteins. We show that divalent cations (e.g. Ca2+, Mg2+ and Mn2+) bind directly to 14-3-3s, and as a result cause a conformational change, manifested as an increase in surface hydrophobicity. A similar change is also obtained by decreasing the pH from pH 7.5 to pH 6.5, in the absence of divalent cations, and we propose that protonation of amino acid residues brings about a similar effect to metal ion binding. A possible regulatory mechanism, where the 14-3-3 protein has to be "primed" prior to binding a target protein, is discussed. PMID- 9871367 TI - Changes in protein interactions of cell cycle-related genes during the dormancy to-growth transition in pea axillary buds. AB - Apical dominance is a phenomenon in which a terminal bud grows predominantly and the growth of the axillary buds is suppressed. Here, we investigated the molecular mechanisms associated with cell cycle control that occur in pea axillary buds as a result of decapitation. Proliferating cell nuclear antigen (PCNA) protein was detected in both dormant and growing buds, while PCNA mRNA was absent in dormant buds. Pissa; CycB1;2 and Cdc2 proteins were undetectable during dormancy. To analyze an interaction between PCNA and Pissa;CycD3;1, we performed anti-PCNA immunoaffinity column chromatography. Pissa;CycD3;1 protein was detected in the eluate prepared from the dormant buds, but not in the eluate prepared from the growing buds. Furthermore, we performed anti-Pissa;CycD3;1 immunoaffinity column chromatography. PCNA protein was detected in the eluate prepared from the dormant buds, but not in the eluate prepared from the growing buds. These results indicated that PCNA associated with Pissa;CycD3;1 only during dormancy. In addition, the interaction between PCNA and Pissa;CycD3;1 was confirmed by a yeast two-hybrid system. PMID- 9871368 TI - Somatic and zygotic embryos of Daucus carota L. display different protein patterns until conversion to plants. AB - Total protein patterns of different developmental stages of carrot zygotic and somatic embryos revealed by one- and two-dimensional gel electrophoresis were compared using statistical dissimilarity index matrix, and some major polypeptides were partially sequenced. In spite of similar morphology, the protein patterns of somatic embryos at the torpedo stage were clearly different from those of zygotic embryos. In particular, none of the proteins specific of zygotic embryos required for maturation, previously identified, were accumulated in somatic embryos, namely the daucin (a globulin-type storage protein), the RAB25 protein (a late embryogenesis abundant protein) (Dodeman et al. 1998), as well as a novel globulin of M(r) 30,000, that we proposed to name apiacin. Somatic plantlets and seedlings also showed different patterns. This discrepancy likely reflects culture conditions, since somatic embryos recover a protein pattern close to that of seedlings after conversion to plant and growth on a carbon-free medium. PMID- 9871370 TI - Infants selectively encode the goal object of an actor's reach. AB - Research with young children has shown that, like adults, they focus selectively on the aspects of an actor's behavior that are relevant to his or her underlying intentions. The current studies used the visual habituation paradigm to ask whether infants would similarly attend to those aspects of an action that are related to the actor's goals. Infants saw an actor reach for and grasp one of two toys sitting side by side on a curtained stage. After habituation, the positions of the toys were switched and babies saw test events in which there was a change in either the path of motion taken by the actor's arm or the object that was grasped by the actor. In the first study, 9-month-old infants looked longer when the actor grasped a new toy than when she moved through a new path. Nine-month olds who saw an inanimate object of approximately the same dimensions as the actor's arm touch the toy did not show this pattern in test. In the second study, 5-month-old infants showed similar, though weaker, patterns. A third study provided evidence that the findings for the events involving a person were not due to perceptual changes in the objects caused by occlusion by the hand. A fourth study replicated the 9 month results for a human grasp at 6 months, and revealed that these effects did not emerge when infants saw an inanimate object with digits that moved to grasp the toy. Taken together, these findings indicate that young infants distinguish in their reasoning about human action and object motion, and that by 6 months infants encode the actions of other people in ways that are consistent with more mature understandings of goal-directed action. PMID- 9871369 TI - Agr, an Agravitropic locus of Arabidopsis thaliana, encodes a novel membrane protein family member. AB - Mutations in the Agr locus of Arabidopsis thaliana impair the root gravitropic response. Root growth of agr mutants is moderately resistant to ethylene and to an auxin transport inhibitor. Vertically placed agr roots grow into agar medium containing IAA or naphthalene-1-acetic acid, but not into medium containing 2,4 D. Positional cloning showed that AGR encodes a root-specific member of a novel membrane-protein family with limited homology to bacterial transporters. PMID- 9871371 TI - Subjectless sentences in child Danish. AB - Three alternative accounts of subject omission, pragmatic, processing and grammatical, are considered from the perspective of child Danish. Longitudinal data for two Danish children are analyzed for subject omission, finite and infinitival verb usage and discourse anchorage of sentence subjects (overt and missing). The data exhibit a well-defined phase of subject omission which coincides with a well-defined phase of infinitival verbal utterances. No evidence is found for input driven accounts of subject omission. Danish adults rarely omit subjects from utterance initial position. Neither is there any evidence to support the claim that omitted subjects are anchored in previous discourse. Evidence supporting a processing constraint explanation of missing subjects is equivocal. The pattern of subject omission, infinitival usage and third person pronoun and past tense usage points to a grammatical explanation of the phenomenon. However, current grammatical accounts have difficulty accommodating several aspects of the data reported. Contrary to structure building theories, the Danish children do not exhibit a phase of development where only uninflected verb forms are used. Danish children also omit subjects from finite utterances. Furthermore, the decline of subject omissions in finite utterances coincides with decline in usage of infinitival utterances. These findings challenge tense-based accounts of children's subject omission. Finally, Danish children exhibit an asymmetry in subject omission according to verb type; subjects are omitted from main verb utterances more frequently than from copula utterances. Given the language typology associated with Danish, this asymmetry is difficult to accommodate within truncation and tense or number-based accounts of subject omission. We suggest that a proper treatment of child subject omission will involve an integration of grammatical and discourse-based approaches. PMID- 9871372 TI - Childhood memory and self-description in young Chinese adults: the impact of growing up an only child. AB - This study examined the relationship between self-description and childhood memory in 255 Chinese young adults. Ninety-nine participants were from only child families and 156 had siblings. All participants completed two questionnaires: a version of the Twenty Statements Test of Kuhn and McPartland (Kuhn, M.H., McPartland, T.S., 1954. An empirical investigation of self-attitudes. American Sociological Review 19, 68-76) eliciting self-descriptions, and an instrument asking for earliest and other childhood memories. Based on theories positing a relationship between autobiography and the organization of the self, we predicted differences on both measures between only- and sibling-child participants. Findings indicated that compared with sibling children, only children had more private and fewer collective self-descriptions, earlier first memories, more specific and more self-focused memories. In addition, autobiographical measures were influenced by cohort, gender, preschool attendance, and urban/rural family effects. Findings are discussed in terms of literature on autobiography, the self and childhood in China. PMID- 9871373 TI - Effects of physical connectivity on the representational unity of multi-part configurations. AB - While physical connection strongly supports perceptual integration of visual elements or parts into single objects, concavities along an object's contour strongly induce segmentation of objects into different parts. By means of a matching task, we examine whether the segmentation of connected parts and the grouping of disconnected parts imply differences in representational unity. All match trials consisted of either two identical configurations or two 180 degrees rotated versions of an otherwise identical configuration, whereas mismatch trials had one misoriented part. Results show that the 180 degrees matches with the physically disconnected parts were relatively slower than similar matches with physically connected parts, although the connections could be minimal (deep concavities, corner to edge connection). This suggests that the mental operation required for the 180 degrees match has more difficulty with the disconnected parts, indicating a measurably lower degree of representational unity. PMID- 9871374 TI - Conditional reasoning with inducements and advice. AB - In an earlier study of conditional reasoning, Newstead et al. [Newstead, S.E., Ellis, C.E., Evans, J.St.B.T., Dennis, I., (1997). Conditional reasoning with realistic material. Thinking and Reasoning 3, 49-96] found that people drew more inferences from conditionals framed as inducements (threats and promises) than from conditionals phrased as advice (tips and warnings). The present study was designed to test the hypothesis that this difference arose from the fact that the speaker of an inducement is normally seen to have control over the consequent event whereas the giver of advice does not. In the experiment reported here, inducement and advice conditionals were constructed in brief contexts such that in either case the speaker could be seen to have high or low control. Participants drew many more conditional inferences of all kinds for high control than for low control conditionals in either context. A second finding of interest was that participants drew many more forward (antecedent to consequent) inferences than backward inferences with these kinds of realistic conditionals. PMID- 9871375 TI - Reproductive health care. A case for uniform prescriptive privileges. PMID- 9871376 TI - Breast health care. A review. AB - Breast health care was rarely acknowledged in the health and science fields prior to this century and has only begun, in recent years, to receive attention outside of pregnancy/lactation or cancer screening and treatment. Yet much health care is involved with regard to this reproductive and sexual organ. With any group of clients, practitioners of women's health care must address an assortment of breast health matters. This article offers an overview of the history of breast health care, the epidemiology of benign breast conditions and cancer, the anatomy and physiology of the breast, and breast development over the lifespan. Also presented are a review of breast assessment and examination, suggestions for routine care of the breast, and an overview of major health issues related to this reproductive organ. Health issues addressed include, among others, nodular and cystic changes, nipple discharge, breast/nipple pain, mastitis, elective alterations, and cancer. Discussion is focused on the latest approaches to optimal breast health care. PMID- 9871377 TI - Sexually transmitted diseases and reproductive health in women. AB - The effect of sexually transmitted diseases (STDs) on the reproductive health of women, the reasons why women bear a disproportionate percent of STD morbidity and mortality, and the factors contributing to an increased risk for STDs and their complications in women are enumerated. Conclusions and recommendations from the 1997 Institute of Medicine report, The Hidden Epidemic: Confronting Sexually Transmitted Diseases, are categorized and addressed. The categories are education, population groups, and clinical practice. The contributions of midwives to implementing pertinent Institute of Medicine recommendations in basic and continuing education programs, individual practice, and in influencing policy are detailed. Factual information about STDs, risk assessment history, essential STD-related services, and treatment guidelines are provided in tables. PMID- 9871378 TI - Preconception care. An integral part of primary care for women. AB - Preconception health care should be an integral part of primary health care services for all women with the potential for childbearing. This article delineates those aspects of preconception care that should be included in a well woman visit. Specific interventions for risks identified in the preconception health care visit are reviewed. Finally, the importance of preconception health care in reducing poor perinatal outcome and promoting overall good health is discussed. PMID- 9871379 TI - Nutrition in reproductive health. AB - This article reviews nutrition-related issues affecting women and their reproductive health. Health care providers must be able to perform a basic nutritional assessment to identify risk factors and develop a plan of care to reduce those risk factors and improve health. Guidelines are provided to assist in performing a nutritional status assessment. Nutritional assessment of women of reproductive age should identify factors that may affect fertility, periconceptional health, and pregnancy outcome. Recommendations are provided to assist the health care provider in counseling women regarding the relationship of food choices and exercise to health, fitness, and optimal bodily function. Controversies surrounding the effect of micronutrient deficits and excesses on reproduction and correction for these imbalances are discussed. Women should be encouraged to initiate dietary and other lifestyle changes to allow for optimal reproductive outcomes. PMID- 9871380 TI - Contraceptive choice and patient compliance. The health care provider's challenge. AB - Contraceptive compliance is a multifaceted issue that is influenced by many factors. These factors can directly affect the level of patient compliance, thereby affecting contraceptive method efficacy rates. A review of the literature reveals many studies about contraceptive compliance but a dearth of studies addressing how to change noncompliant behaviors. This article describes the contraceptive methods currently available and their efficacy rates. Patient characteristics and the components of compliance are described as they affect contraceptive efficacy and patient care. Suggestions are made for the use of alternative terminology to include adherence to or continuance of a contraceptive method. Health care providers should realize the impact they can have on a patient's education, decision-making process, and ultimate compliance with a contraceptive method. It is the patient, however, who ultimately makes the decision, either actively or passively, to comply or not and whether to have an unplanned pregnancy. PMID- 9871381 TI - Unintended pregnancy. Consequences and solutions for a worldwide problem. AB - Unintended pregnancy is a worldwide problem that affects women, their families, and society. Unintended pregnancy can result from contraceptive failure, non-use of contraceptive services, and, less commonly, rape. Abortion is a frequent consequence of unintended pregnancy and, in the developing world, can result in serious, long-term negative health effects including infertility and maternal death. In many developing countries, poverty, malnutrition, and lack of sanitation and education contribute to serious health consequences for women and their families experiencing an unintended pregnancy. Regardless of the cause, unintended pregnancy and its negative consequences can be prevented by access to contraceptive services including emergency contraception, safe and legal abortion services, and a society that allows women to determine their own reproductive choices. Addressing unintended pregnancy and its substantial human and dollar costs should be a priority in every country. The availability of reliable contraception for all, regardless of age or ability to pay, is an essential first step. Women and adolescents require access to age-appropriate and culturally sensitive reproductive health care services, including emergency contraception. Access to safe, legal abortion services is necessary to impact the staggering maternal mortality rates worldwide. Midwives throughout the world provide the majority of care for women of reproductive age. It is essential to identify those at risk for unintended pregnancy, provide the services they require, and remain diligent to ensure that those women and their families have safe options to consider when faced with an unintended pregnancy. In 1920, Magaret Sanger said, "No women can call herself free who does not control her own body." Although great strides have been made to improve the health and status of women since Ms. Sanger spoke those words, there remains much work to be done. PMID- 9871382 TI - Early abortion. Update and implications for midwifery practice. AB - Medical abortion using methotrexate and misoprostol and manual vacuum aspiration are two new methods for pregnancy termination during the first 8 weeks of gestation. Compared to the regimen of mifepristone (RU 486) and misoprostol, both methods offer high rates of complete abortion and acceptability to users. Limitations of the new two-drug regimen compared with mifepristone include a longer time to effect abortion, transient gastrointestinal side effects, and risk of potential teratogenicity from methotrexate's cytotoxicity. Compared to standard surgical abortion, both methods allow women to avoid surgery, are more privately performed, and may be more easily accessible. The safety of first trimester abortion provided by nurse practitioners and physician assistants has been established. Whether midwives and either new method to their practices depends on several factors. These include obtaining appropriate training, overcoming legal restrictions, and meeting professional and personal challenges inherent in providing early abortion care. PMID- 9871383 TI - Current issues in the midwifery management of women living with HIV/AIDS. AB - Changes in the clinical management of women living with human immunodeficiency virus (HIV) have occurred as a result of significant gains in the scientific knowledge of this retrovirus. As the incidence of HIV/AIDS continues to escalate among female adults and adolescents in the United States, all primary health care providers must anticipate the likelihood of encountering clients with HIV infection. Midwives must be adequately prepared to meet the challenges of managing women with HIV in the early stages of the disease. This article presents a comprehensive review of current demographic trends related to the HIV/AIDS epidemic among U.S. women and a brief overview of the essential immunopathogenesis of HIV. Contemporary issues related to universal counseling, updated testing procedures, and reproductive decision-making are covered. Initial primary care concerns and the management of newly infected seropositive women are included, with a focus on gynecologic issues. Guidance in the current management of HIV-positive pregnant women is offered. Updated antiretroviral prophylaxis recommendations are presented, to prevent perinatal transmission and to delay maternal immunosuppression with subsequent opportunistic infections. The article concludes with implications for health care professionals who provide care for this unique cohort of women. PMID- 9871384 TI - History of nurse-midwifery in reproductive health care. AB - From the maternity cycle to the primary health care of women, this article traces the expansion of the role of midwives in reproductive health care over the past 3 decades. Included are the recollections of Shirley Okrent about the early development of this role expansion and a few of the author's personal memories. PMID- 9871385 TI - Opportunities and burdens for nurse-midwives working in primary health care. An example from population-based cervical cancer screening in urban Sweden. AB - In Sweden, nurse-midwives in primary health care are responsible for taking Papanicolaou smears in population-based cervical cancer screening programs. This article discusses the manner in which a group of nurse-midwives, working with the cervical cancer screening program, view both opportunities and burdens inherent in their work. Semistructured interviews were conducted with 21 nurse-midwives engaged in screening at seven antenatal health care centers in demographically different parts of Stockholm, using a team approach to collect and inductively analyze the interviews. Results indicate discrepancies between ideals guiding the midwives and their practice. Positive aspects described by the midwives appear to be related to an ideology of care, whereas perceived burdens include the midwives' sense of powerlessness, lack of congruence between midwives' ideology and screening organization, and lack of professional familiarity with cancer. Several of these features can be related to issues that are challenges for many health care practitioners today, even in other settings. Our conclusion is that the screening program has latent potential for further development of the role of the nurse-midwife as an advocate for women throughout the lifespan and in a variety of situations. PMID- 9871386 TI - Emergency contraception (ECP) PMID- 9871387 TI - The only answer to the dilemma of the health care industry: total emphasis on rural health care in America. PMID- 9871388 TI - Medicare launches pilot of new beneficiary complaint alternatives--testing to take place in six states. PMID- 9871389 TI - Community solutions for access to medical care. PMID- 9871390 TI - Arkansas Health Care Access Foundation begins its tenth year of helping Arkansas in need. PMID- 9871391 TI - Measuring the success of the Arkansas Health Care Access Foundation. AB - This report is an attempt to measure the effectiveness of the Health Care Access Program and to study the demographic elements of the patient population served. The study involved a two-pronged approach. By polling both physicians and patients regarding their experiences with AHCAF, we can form conclusions about its effectiveness. In addition, demographic information regarding patient make-up and physician participation in regional areas can serve as a resource for strategic management of the program. PMID- 9871392 TI - Trauma deaths in a rural area. AB - This study was undertaken in order to clarify the causes of death amongst rural trauma victims in Union County, Arkansas. Over a five-year time interval, seventy two percent (n = 60) of the trauma deaths occurred at the geographical site of injury before the arrival of any emergency medical services. Thus, prolonged time to discovery of trauma victims accounts for the most significant factor leading to trauma deaths in Union County. Education regarding injury prevention must continue to be emphasized to the public. PMID- 9871393 TI - Community Health Centers: part of the health care safety net. PMID- 9871394 TI - Latex allergies. PMID- 9871395 TI - The dentition of Abraham's people. Why Abraham left Mesopotamia. AB - Ancient Mesopotamia (southern Iraq) in 2000 B.C. was the home of the biblical figure Abraham and his people. An analysis of skeletal material from cemeteries in the ancient cities of Ur (birthplace of Abraham) and Kish shows a homogeneous, short-lived population that suffered severe dental attrition (95 percent of inhabitants), periodontal disease (42 percent) and caries (two percent). Many oral congenital and neoplastic lesions were noted Skeletal (dental) evidence indicates the population suffered from extensive disease and nutritional deficiency. Of special note was a possible case of endodontic treatment. The people of Abraham had modern dentition but relatively poor dental health. It is probable that Abraham, if in truth he existed, left Mesopotamia because of the threat of famine, as evidenced by biblical record, geologic strata and dental analysis. PMID- 9871396 TI - Outpatient anesthesia in oral and maxillofacial surgery. AB - An integral part of the specialty of oral and maxillofacial surgery is intra operative anxiety and pain control with the use of inhalation and intravenous agents. The safety of these agents when used by trained practitioners and staff in properly equipped facilities is well-documented. The purpose of this article is to review the most common sedative agents currently used in the oral and maxillofacial surgery outpatient setting. PMID- 9871397 TI - Plexiform ameloblastoma involving the maxillary antrum. A case report. AB - There have been only seven cases of plexiform ameloblastoma involving the maxillary sinus reported in the English literature. The eighth case is reported here. It involves the maxillary antrum of an 80-year-old female. PMID- 9871398 TI - ADA reacts to laser approval. PMID- 9871399 TI - Surgical landmarks during mastoidal and petrosal operations. AB - This study was made on twenty-one formalin fixed, adult skull left-half specimens. Each of the measurements was made using callipers accurate to 0.1 mm. The dissection of the temporal bone was begun with simple mastoidectomy and completed when the endosteum of the semicircular canals were opened. During the temporal bone dissection step by step, eleven different measurements have been made with small size callipers, and mean, median, range and standard deviations have been determined. The following set of correlations was found to be significant; I--The correlation between the orbitomeatal length and the distance from the most lateral point of the tympanomastoid suture to the facial canal (r = 0.69, p < 0.001). II--The correlation between the orbitomeatal length and the distance from the tympanic ring to the facial canal (r = 0.49, p = 0.011). III- The correlation between the distance from the most lateral point of the tympanomastoid suture to the facial canal and the distance from the tympanic ring to the facial canal (r = 0.71, p < 0.001). PMID- 9871400 TI - Occlusion of the femoral artery induced fos-like immunoreactive neurons in the lateral habenular nucleus projecting to the midbrain periaqueductal gray in the rat. AB - Wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) injection into the midbrain periaqueductal gray (PAG) resulted in heavy accumulation of retrogradely labeled neurons in the lateral habenular nucleus (LHb) bilaterally. When the left femoral artery was persistently ligated for 2 h, the expression of Fos-like immunoreactivity (FOS-LI) was also found bilaterally in the LHb. In the present study, by combining the retrograde labeling method of injecting WGA-HRP into the PAG and the immunohistochemical staining of the FOS-LI neurons in the LHb induced by occlusion of the femoral artery, it was demonstrated that there are neurons containing both HRP labeling and FOS-LI. These neurons appeared to be located mainly in the medial part and posterior half of the LHb, and constitute about 24% of the total number of all labeled cells. PMID- 9871401 TI - What is anatomy for? Considerations on the body as a whole and whole anatomy. AB - In anatomy today, construction of the body is devoid of the water element, and the framework of the discipline is sectionalized (segmented, specialized, and divided), so that the body is not described in an integrated, harmonious manner. In this paper, the present authors selected the body, body water, and skin as objects of contemplation and deliberated on the history, significance, and resuscitation of anatomy. (1) Living things are dependent on water, and water is a vital component of the body. Therefore, greater consideration should be given to the presence of body water in morphological description of the body. (2) Since anatomy is sectionalized, understanding of the body is dwarfed and distorted. Whole anatomy, which described the body as a whole is the ideal style. (3) The body as a whole should always be placed in the center of anatomy. This should be the ultimate form of teaching anatomy. PMID- 9871402 TI - Expressions of a 68kDa-glycoprotein (GP68) and laminin in the mesodermal tissue of the developing mouse embryo. AB - Immunohistochemistry revealed initial expression of the stage-specific glycoprotein, GP68, in various mesenchymal tissue substructures of mouse embryos. During the 11-15th days of gestation, GP68 was localized in the primitive meninges, chondroblasts and perichondrium of pre-cartilaginous vertebral bodies and ribs, connective tissue cells of the dermis, the epicardium and endocardium of the heart, the epimysium and perimysium of skeleton musclature, and the basement membranes of splanchnic organs. Double staining for laminin expression indicated coincidental expression in identical tissue substructures. However, laminin was expressed in days 10-18 embryos and the neonate. Therefore, GP68 is coincidentally expressed with laminin in mesenchymal tissues between the 11th and 15th day of gestation, and may play a role as a laminin-associated protein. In the light of these results, a hypothesis concerning the relationship between these two proteins and the mechanisms of non-integrin laminin-associated proteins during normal embryogenesis is discussed further. PMID- 9871403 TI - The endomysium of human embryonic hyoid muscle during development. AB - A computer analysis was performed on the endomysium of the hyoid muscle in human fetuses, which ranged in gestational week from 12 to 32 weeks. During development of the human hyoid muscle a level of intramuscular connective tissue as reticular fibers displays a specific change in morphology. We examined quantitates and distributions of endomysium in human hyoid muscles from 12 to 32 weeks. The endomysium of the hyoid muscles formed complex structures, and volume rates increased from 20 weeks gestation. On the other hand, the cross sectional area (CSA) of muscle fibers had almost the same profile. The reticular fibers increased during development from 16 to 28 weeks gestation. PMID- 9871404 TI - Immunocytochemical study of the maxilla and maxillary sinus during human fetal development. AB - We quantitatively examined the distribution of the proliferating cell nuclear antigen (PCNA), which is associated with cell division, and of components of the ECMs (collagen types I and III, tenascin and osteonectin) in the immature zones at three sites: palatine, inferior and vertical surface region of the fetal human maxilla as well as the maxillary sinus (MS) at 12, 16, 20, 24, and 28 weeks gestation. The percentage of PCNA-positive cells was the highest at 16 weeks in the immature zones of maxilla. Tenascin and fibrillar collagens (collagen types I and III) were especially present in the cellular zone linked to the bone and cartilage matrices of the immature zones of the maxillary bone at 20-24 weeks gestation. The osteonectin was detected on the maxillary bone from 24 weeks. These distributions of ECMs revealed the specific and contrasting profiles of development in the human maxillary bone and might reflect the formation of paranasal sinus as MS. PMID- 9871405 TI - Is the free androgen index a useful clinical marker in male patients? AB - The clinical relevance of the free androgen index (FAI), a ratio of total testosterone (T) to sex-hormone binding globulin (SHBG), was investigated in a regional population of men (n = 40) and women (n = 30). The FAI correlated well with free testosterone (T) in both men (r = 0.551, p < 0.001) and women (r = 0.454, p < 0.01). However, there was considerable variability among individual patients. Moreover, the FAI showed no association with sperm parameters in male patients, although total T and free T showed weak associations. The FAI may be a cost-effective alternative to free T measurement in the diagnosis of oligomenorrhea and hirsutism in women as previously shown, but may have little relevance in men. PMID- 9871406 TI - ATP-sensitive K+ channels come of age. PMID- 9871407 TI - Modulation of intracellular Ca2+ levels in the heart by sorcin and FKBP12, two accessory proteins of ryanodine receptors. PMID- 9871408 TI - Isoforms of endothelin-converting enzyme: why and where? PMID- 9871409 TI - The use of nonhomologous scatchard analysis in the evaluation of ligand-protein interactions. AB - Scatchard plots are widely used for the graphical presentation of receptor-ligand binding data. When a combination of labelled and unlabelled ligand molecules is used in a binding assay, equations for Scatchard plots are readily available if the labelled and unlabelled ligands have similar binding affinities. In this article, Everardus van Zoelen, Roel Kramer, Herman van Moerkerk and Jacques Veerkamp present mathematical equations to obtain the binding characteristics of an unlabelled ligand in a Scatchard plot, which has a dissociation equilibrium constant different from that of the labelled ligand used. PMID- 9871410 TI - Ethanol tolerance and synaptic plasticity. AB - Current concepts of the mechanisms underlying many of the pharmacological effects of ethanol on the CNS involve disruption of ion channel function via the interaction of ethanol with specific hydrophobic sites on channel subunit proteins. Of particular clinical importance is the development of tolerance and dependence to ethanol, and it is likely that adaptive changes in synaptic function in response to ethanol's actions on ion channels play a role in this process. In this article, Judson Chandler, Adron Harris and Fulton Crews discuss potential mechanisms of ethanol-induced changes in synaptic function that might provide a cellular basis for ethanol tolerance and dependence. It is proposed that multiple mechanisms are involved that include both transcriptional and post translational modifications in NMDA and GABAA receptors. PMID- 9871411 TI - Glucose action 'beyond ionic events' in the pancreatic beta cell. AB - For normal glucose homeostasis, insulin release by the pancreatic beta cell is vital. Until recently, it was thought that glucose-induced ionic events, such as closure of the ATP-sensitive K+ (KATP) channels, membrane depolarization, activation of the L-type voltage-dependent Ca2+ channels, Ca2+ influx and elevation of cytosolic free Ca2+, constitute the main signalling pathway in beta cell stimulus-secretion coupling. However, since the discovery of 'non-ionic' glucose actions in the beta cell by the Aizawa and Henquin laboratories in 1991, data have accumulated that strongly indicate the physiological relevance of this signalling pathway. In this review, Toru Aizawa and colleagues discuss how the KATP channel-Ca2+ hypothesis was formulated, what was overlooked in the hypothesis, and then provide a comprehensive view of stimulus-secretion coupling in the beta cell, with an emphasis on non-ionic glucose actions. PMID- 9871412 TI - Two isoforms of glutamate decarboxylase: why? AB - Adults express two isoforms of glutamate decarboxylase (GAD), GAD67 and GAD65, which are encoded by different independently regulated genes, a situation that differs from that of other neurotransmitters. In this article, J-J. Soghomonian and David Martin review current knowledge on the differences between these two isoforms. Both isoforms are present in most GABA-containing neurones in the CNS, but GAD65 appears to be targeted to membranes and nerve endings, whereas GAD67 is more widely distributed in cells. Both forms can synthesize transmitter GABA, but GAD67 might preferentially synthesize cytoplasmic GABA and GAD65 might preferentially synthesize GABA for vesicular release. Several lines of evidence suggest that the two forms have different roles in the coding of information by GABA-containing neurones. PMID- 9871413 TI - Metabotropic receptors for ATP and UTP: exploring the correspondence between native and recombinant nucleotide receptors. AB - In the past five years, an extended series (P2Y1-n) of metabotropic nucleotide (P2) receptors has been cloned from vertebrate tissues; these receptors are activated by either ATP or UTP, or both nucleotides. While certain cloned P2Y receptors appear to correspond functionally to particular native P2 receptor phenotypes, such pharmacological phenotypes could be explained by either a combination of several members of the P2Y1-n series being coexpressed in the same tissue or the existence of novel, uncloned P2Y subtypes. Here, Brian King, Andrea Townsend-Nicholson and Geoffrey Burnstock review recent findings on the matter of pharmacological relationships between native P2 and cloned P2Y receptors. PMID- 9871414 TI - A calling to care. Parish nurses in Richmond, Va., place a lot of faith in their healing mission. PMID- 9871415 TI - The year that was--what it means for 1999. PMID- 9871416 TI - It serves you right. Hassle-free health care builds loyalty, volume--and the bottom line. AB - Zero barriers to care, zero waiting times: Tough as those targets sound, today's stars of customer service are scoring direct hits. "It's not a change in the services that are delivered," says one patient satisfaction pro. "It's a change in the way they're being delivered." PMID- 9871417 TI - Yes. No. Maybe. That's about as firm as the guidance gets over launching a provider-sponsored health plan. Strategy roundtable. AB - Few participants in our roundtable on provider-sponsored health plans see them the same way. Some are gearing up new products, others are scaling back or getting out. Yet they agreed on one thing: The financial risks are tremendous. PMID- 9871418 TI - Direct contracts. Running for coverage. PMID- 9871419 TI - Treatment turf. A piece of pain relief. PMID- 9871420 TI - Customer service. Worthier waits. PMID- 9871421 TI - Fringe benefits. More seance than science? PMID- 9871422 TI - Quackery. Muddled miracles. PMID- 9871423 TI - The effectiveness of antihistamines in reducing the severity of runny nose and sneezing: a meta-analysis. PMID- 9871424 TI - Thalidomide does not alter the pharmacokinetics of ethinyl estradiol and norethindrone. AB - OBJECTIVE: To evaluate the effect of thalidomide on the plasma pharmacokinetics of ethinyl estradiol (INN, ethinylestradiol) and norethindrone (INN, norethisterone). METHODS: Ten women who had undergone surgical sterilization were enrolled in an open-label crossover study conducted in the Georgetown University Clinical Research Center. The pharmacokinetics of single doses of 0.07 mg ethinyl estradiol and 2 mg norethindrone were measured at baseline and after 3 weeks of 200 mg thalidomide. Compliance with the thalidomide regimen was assessed with use of Medication Event Monitoring System (MEMS) caps. RESULTS: No changes were observed in the pharmacokinetics of ethinyl estradiol or norethindrone with thalidomide therapy. The mean +/- SD area under the plasma concentration-time curve (AUC0-infinity) for ethinyl estradiol was 6580 +/- 1100 ng.h/L at baseline and 5970 +/- 1560 ng.h/L after the thalidomide regimen (paired t test, P > .05). The values for norethindrone were 103 +/- 54 micrograms.h/L and 107 +/- 58 micrograms.h/L (paired t test, P > .05). No changes were observed for other pharmacokinetic parameters assessed for either ethinyl estradiol or norethindrone. No accumulation of thalidomide was seen after 21 days of therapy: day 1 AUC0-infinity 41.1 +/- 13.9 micrograms.h/mL; day 21 AUC0-infinity 59.6 +/- 27.3 micrograms.h/mL (paired t test, P > .05). No changes were observed for other pharmacokinetic parameters assessed for thalidomide between days 1 and 21. Thalidomide was well tolerated but caused variable degrees of sedation. The average thalidomide compliance rate was 97%. CONCLUSIONS: The pharmacokinetics of thalidomide do not change with 3 weeks of daily dosing. Thalidomide does not alter the pharmacokinetics of ethinyl estradiol or norethindrone. Therefore there is no drug interaction between thalidomide and these 2 drugs. The efficacy of oral contraceptives containing ethinyl estradiol and norethindrone should not be affected by concomitant thalidomide therapy. PMID- 9871425 TI - Effects of blocking CYP2D6 on the pharmacokinetics and pharmacodynamics of oxycodone. AB - BACKGROUND: Oxycodone is metabolized in the liver by means of O-demethylation to form oxymorphone in a reaction catalyzed by the enzyme cytochrome P450 2D6 (CYP2D6). This enzyme is expressed as 2 phenotypes (extensive and poor metabolizers). Several drugs are metabolized by CYP2D6, and clinically relevant drug interactions may occur. The aim of this study was to evaluate the role of oxymorphone in mediating the opioid effects of oxycodone by means of blocking CYP2D6 with quinidine. METHODS: Ten healthy extensive metabolizers were administered 20 mg controlled-release oxycodone after premedication with placebo or 200 mg quinidine in this randomized, double-blind crossover study. A dose of 100 mg quinidine was administered 6 hours later. Plasma opioid concentrations, subjective pharmacodynamic ratings, and psychomotor function were assessed for 24 hours after drug administration. RESULTS: No oxymorphone was detected at any time after quinidine premedication in 8 of 10 subjects. Plasma oxycodone (difference not significant) and noroxycodone (P < .01) concentrations were greater after quinidine pretreatment. Prevention of the production of oxymorphone by quinidine did not affect the psychomotor or subjective drug effects of oxycodone. No difference in number of adverse effects was observed after the 2 pretreatments. CONCLUSIONS: A significant reduction in plasma oxymorphone levels did not substantially alter the pharmacodynamic effects of oxycodone. Analgesia was not evaluated because pain was not present. PMID- 9871426 TI - Pharmacokinetics, pharmacodynamics, and tolerance of single- and multiple-dose fexofenadine hydrochloride in healthy male volunteers. AB - BACKGROUND: Fexofenadine is a selective, nonsedating H1-receptor antagonist that relieves symptoms of allergic conditions. METHODS: Two randomized, double-blind, parallel-group, placebo-controlled dose-escalation studies were performed in healthy men to determine the maximum tolerated oral dose, pharmacokinetics, pharmacodynamics, and safety of fexofenadine hydrochloride. In the first study, 87 subjects (6 in the active drug group and 2 in the placebo group) received single oral doses of fexofenadine hydrochloride ranging from 10 to 800 mg or placebo. In the second study, 32 subjects (3 in the active drug group and 1 in the placebo group) received multiple fexofenadine hydrochloride doses ranging from 20 to 690 mg or placebo twice daily for 28 1/2 days. Serial plasma and urine samples were collected. Fexofenadine concentrations were determined by HPLC and fluorescence. Wheal and flare response to intradermal histamine was used to evaluate antihistaminic activity. RESULTS: Fexofenadine hydrochloride was rapidly absorbed, reaching peak concentrations in 0.83 to 1.33 hours. Single-dose mean concentration ranged from 46 to 6383 ng/mL, and steady-state maximum plasma concentration ranged from 58 to 4677 ng/mL. Mean area under the plasma concentration-time curve was approximately proportional to dose. Oral clearance, renal clearance, and cumulative percent of drug excreted in urine were similar after single and multiple doses and were generally constant over the dose range studied. Inhibition of skin wheal and flare was shown for single doses of 40 mg and higher and for all multiple doses. No fexofenadine dose-related trends or apparent differences from placebo were found for any safety parameter. CONCLUSIONS: Fexofenadine hydrochloride was well tolerated at oral doses up to 11 times the recommended therapeutic dose. In addition, fexofenadine hydrochloride showed significant antihistaminic activity and dose-proportional pharmacokinetics over a wide dosing range. PMID- 9871427 TI - Population pharmacokinetic-pharmacodynamic modeling of moxonidine using 24-hour ambulatory blood pressure measurements. AB - OBJECTIVES: To develop a model for 24-hour ambulatory blood pressure measurements (ABPM) that can be applied in a pharmacokinetic-pharmacodynamic model. METHODS: Four different data sets were prepared from 2 studies to accommodate different modeling strategies. In study A, a double-blind placebo-controlled study in 47 patients, 24-hour ABPM profiles (74 to 99 measurements per profile) were obtained during the placebo run-in phase and after 3, 5, and 11 weeks during the treatment. Three to 5 plasma samples were taken. Cosine and polynomial models were evaluated to describe the circadian rhythm in blood pressure based on 3 data sets (1: only run-in data; 2: only placebo data; 3: all data). In study B, a double-blind placebo-controlled study in 94 patients, two 24-hour ABPM profiles per patient (during placebo run-in and after 8 weeks) were recorded and randomly reduced to 15 measurements per profile to evaluate the robustness of the baseline model. RESULTS: The mean moxonidine clearance was 35 L/h, and the volume of distribution was 132 L. The final baseline model consisted of 2 cosine terms with fixed-effect parameters for rhythm-adjusted 24-hour mean blood pressure, amplitude, phase, and period; random-effect parameters for interindividual variability in rhythm-adjusted 24-hour mean, amplitude, and clock time; and interoccasion variability in rhythm-adjusted 24-hour mean and clock time. The final baseline model was combined with an Emax model for the drug effect. An effect compartment was used (kco = 0.198 h-1). The maximum decrease in diastolic blood pressure (Emax) was 16.7%, and EC50 was 0.945 microgram/L. CONCLUSION: The pharmacokinetic-pharmacodynamic model for 24-hour ABPM can be used to estimate the concentration-effect relationship of antihypertensive drugs. PMID- 9871428 TI - A model for the turnover of dihydrotestosterone in the presence of the irreversible 5 alpha-reductase inhibitors GI198745 and finasteride. AB - OBJECTIVE: To develop a pharmacokinetic-pharmacodynamic model that characterizes the conversion of testosterone to dihydrotestosterone (DHT) by 5 alpha-reductase types 1 and 2 and the irreversible inhibition of 5 alpha-reductase by finasteride, a 5 alpha-reductase type 2 inhibitor and by GI198745 (dutasteride), a potent and specific dual 5 alpha-reductase inhibitor. METHODS: Healthy men (n = 48) received doses of 0.1 to 40 mg GI198745 (n = 4 subjects per dose), 5 mg finasteride (n = 8), or placebo (n = 8) in a parallel-group study. Plasma concentrations of GI198745, finasteride, and DHT were measured frequently up to 8 weeks after dosing. Models were fitted with mixed-effects modeling with the NONMEM program. RESULTS: The pharmacodynamics were well described with a model that accounted for the rates of DHT formation and elimination, 5 alpha-reductase turnover, relative capacity of the 2 5 alpha-reductase isozymes, and the rates of irreversible inhibition of one (finasteride) or both (GI198745) types of 5 alpha reductase. The model indicated that type 2 5 alpha-reductase contributed approximately 80% of plasma DHT. GI198745 was about 3-fold more potent than finasteride on 5 alpha-reductase type 2. Nearly full blockade of both isozymes was achieved at doses of 10 mg or more GI198745, although the potency of this agent on 5 alpha-reductase type 1 was less than on type 2. CONCLUSIONS: A physiologically based model for the turnover and irreversible inhibition of 5 alpha-reductase and for formation and elimination of DHT described the data well. This model helps explain differences in the rates of onset and offset of effect and offers a way to determine the relative potency of the irreversible 5 alpha reductase inhibitors. PMID- 9871429 TI - Tamoxifen and toremifene concentrations in plasma are greatly decreased by rifampin. AB - BACKGROUND: Rifampin (INN, rifampicin) is a potent inducer of cytochrome P450 (CYP) enzymes involved in drug metabolism and therefore causes many drug interactions. METHODS: The effects of rifampin on the pharmacokinetics of tamoxifen (study I) and toremifene (study II) were examined in 2 randomized, placebo-controlled crossover studies. Ten (study I) or 9 (study II) healthy male volunteers took either 600 mg rifampin or placebo orally once a day for 5 days. On the sixth day, 80 mg tamoxifen or 120 mg toremifene was administered orally. Blood samples were collected up to 336 hours after drug administration. RESULTS: Rifampin reduced the area under the plasma concentration-time curve (AUC) of tamoxifen by 86% (P < .001), peak plasma concentration (Cmax) by 55% (P < .001), and elimination half-life (t1/2) by 44% (P < .001). The AUC of toremifene was reduced by 87% (P < .001), Cmax by 55% (P < .001), and t1/2 by 44% (P < .01) with rifampin. During the rifampin phase, the AUC of N-demethyltamoxifen was 38% (P < .001) and the AUC of N-demethyltoremifene was 20% (P < .01) of that during the placebo phase. CONCLUSIONS: Rifampin markedly reduces the plasma concentrations of tamoxifen and toremifene by inducing their CYP3A4-mediated metabolism. Concomitant use of rifampin or other potent inducers of CYP3A4 with tamoxifen and toremifene may reduce the efficacy of these antiestrogens. PMID- 9871430 TI - Grapefruit juice substantially increases plasma concentrations of buspirone. AB - BACKGROUND: Buspirone has a low oral bioavailability because of extensive first pass metabolism. The effect of grapefruit juice on the pharmacokinetics and pharmacodynamics of orally administered buspirone is not known. METHODS: In a randomized, 2-phase crossover study, 10 healthy volunteers took either 200 mL double-strength grapefruit juice or water 3 times a day for 2 days. On day 3, each subject ingested 10 mg buspirone with either 200 mL grapefruit juice or water, and an additional 200 mL was ingested 1/2 hour and 1 1/2 hours after buspirone administration. Timed blood samples were collected up to 12 hours after ingestion, and the effects of buspirone were measured with 6 psychomotor tests up to 8 hours after ingestion. RESULTS: Grapefruit juice increased the mean peak plasma concentration of buspirone 4.3-fold (range, 2-fold to 15.6-fold; P < .01) and the mean area under the plasma buspirone concentration-time curve 9.2-fold (range, 3-fold to 20.4-fold; P < .01). The time of the peak concentration (tmax) of buspirone increased from 0.75 to 3 hours (P < .01), and the elimination half life (t1/2) was slightly increased (P < .01) by grapefruit juice. A significant increase in the pharmacodynamic effects of buspirone by grapefruit juice was seen only in subjective overall drug effect (P < .01). CONCLUSIONS: Grapefruit juice considerably increased plasma buspirone concentrations. The probable mechanism of this interaction is delayed gastric emptying and inhibition of the cytochrome P450 3A4-mediated first-pass metabolism of buspirone caused by grapefruit juice. Concomitant use of buspirone and at least large amounts of grapefruit juice should be avoided. PMID- 9871432 TI - The pharmacokinetic-pharmacodynamic relationship for mycophenolate mofetil in renal transplantation. AB - BACKGROUND: Mycophenolate mofetil, a pro-drug for mycophenolic acid, reduces the likelihood of allograft rejection after renal transplantation. We studied the relationship between mycophenolic acid pharmacokinetics and the likelihood of rejection in a randomized concentration-controlled trial. METHODS: Under double blind conditions, recipients of kidney transplants were followed for evidence of allograft rejection for 6 months. In addition to mycophenolate mofetil, patients received usual doses of cyclosporine (INN, ciclosporin) and corticosteroids. The dose of mycophenolate mofetil (given twice daily) was controlled by feedback, with mycophenolic acid area under the concentration-time curve (AUC) as the controlled variable. Patients were randomly assigned to 1 of 3 target AUC groups. RESULTS: Logistic regression analysis showed a significant (P < .0001) relationship between mycophenolic acid AUC and the likelihood of rejection. High mycophenolic acid values were associated with a very low probability of rejection. An AUC of 15 micrograms.h/mL yielded 50% of maximal achievable efficacy with a 4% change of efficacy for a 1 microgram.h/mL change in AUC at the midpoint of the logistic curve. Exploratory analyses showed other variables (e.g., the maximum observed plasma concentration, predose plasma concentration, and drug dose) had poorer predictive power for the rejection outcome. Bivariate regression confirmed the importance of AUC as a highly predictive variable and showed low predictive value of other variables, once the contribution of AUC had been considered. The characteristic side effects of mycophenolate mofetil therapy appeared related to drug dose but not to mycophenolic acid concentration. CONCLUSIONS: The AUC of mycophenolic acid is predictive of the likelihood of allograft rejection after renal transplantation in patients receiving mycophenolate mofetil. PMID- 9871431 TI - Kinetic and dynamic interaction study of zolpidem with ketoconazole, itraconazole, and fluconazole. AB - BACKGROUND: Azole antifungal agents may impair hepatic clearance of drugs metabolized by cytochrome P450-3A isoforms. The imidazopyridine hypnotic agent zolpidem is metabolized in humans in part by P450-3A, as well as by a number of other cytochromes. Potential interactions of zolpidem with 3 commonly prescribed azole derivatives were evaluated in a controlled clinical study. METHODS: In a randomized, double-blind, 5-way, crossover, clinical pharmacokinetic pharmacodynamic study, 12 volunteers received (A) zolpidem placebo plus azole placebo, (B) 5 mg zolpidem plus azole placebo (C) zolpidem plus ketoconazole, (D) zolpidem plus itraconazole, and (E) zolpidem plus fluconazole. RESULTS: Mean apparent oral clearance of zolpidem when given with placebo was 422 mL/min, and elimination half-life was 1.9 hours. Clearance was significantly reduced to 250 mL/min when zolpidem was given with ketoconazole, and half-life was prolonged to 2.4 hours. Coadministration of zolpidem with itraconazole or fluconazole also reduced clearance (320 and 338 mL/min), but differences compared to the zolpidem plus placebo treatment did not reach significance. Zolpidem-induced benzodiazepine agonist effects (increased electrocardiographic beta activity, digit-symbol substitution test impairment, and delayed recall) during the first 4 hours after dosage were enhanced by ketoconazole but not by itraconazole or fluconazole. CONCLUSION: Coadministration of zolpidem with ketoconazole impairs zolpidem clearance and enhances its benzodiazepine-like agonist pharmacodynamic effects. Itraconazole and fluconazole had a small influence on zolpidem kinetics and dynamics. The findings are consistent with in vitro studies of differentially impaired zolpidem metabolism by azole derivatives. PMID- 9871433 TI - Benzodiazepine use and cognitive function among community-dwelling elderly. AB - OBJECTIVE: To evaluate the relation between benzodiazepine use and cognitive function among community-dwelling elderly. METHODS: This prospective cohort study included 2765 self-reporting subjects from the Duke Established Populations for Epidemiologic Studies of the Elderly. The subjects were cognitively intact at baseline (1986-1987) and alive at follow-up data collection 3 years later. Cognitive function was assessed with the Short Portable Mental Status Questionnaire (unimpaired versus impaired and change in score) and on the basis of the number of errors on the individual domains of the Orientation-Memory Concentration Test. Benzodiazepine use was determined during in-home interviews and classified by dose, half-life, and duration. Covariates included demographic characteristics, health status, and health behaviors. RESULTS: After control for covariates, current users of benzodiazepine made more errors on the memory test (beta coefficient, 0.35; 95% confidence interval [CI], 0.10 to 0.61) than nonusers. Further assessment of the negative effects on memory among current users suggested a dose response in which users taking the recommended or higher dose made more errors (beta coefficient, 0.57; 95% CI, 0.26 to 0.88) and a duration response in which long-term users made more errors (beta coefficient, 0.39; 95% CI, 0.05 to 0.73) than nonusers. Users of agents with long half-lives and users of agents with short half-lives both had increased memory impairment (beta coefficient, 0.32; 95% CI, 0.01 to 0.64 and beta coefficient, 0.38; 95% CI, 0.02 to 0.75, respectively) relative to nonusers. Previous benzodiazepine use was unrelated to memory problems, and current and previous benzodiazepine use was unrelated to level of cognitive functioning as measured with the other 4 tests. CONCLUSIONS: The results suggested that current benzodiazepine use, especially in recommended or higher doses, is associated with worse memory among community dwelling elderly. PMID- 9871434 TI - Dopamine-glutamate interactions in the basal ganglia. AB - In an attempt to formulate a working hypothesis of basal-ganglia functions, arguments are considered suggesting that the basal ganglia are involved in a process of response selection i.e. in the facilitation of "wanted" and in the suppression of "unwanted" behaviour. The meso-accumbal dopamine-system is considered to mediate natural and drug-induced reward and sensitization. The meso striatal dopamine-system seems to fulfill similar functions: It may mediate reinforcement which strengthens a given behaviour when elicited subsequently, but which is not experienced as reward or hedonia. Glutamate as the transmitter of the corticofugal projections to the basal ganglia nuclei and of the subthalamic neurons is critically involved in basal ganglia functions and dysfunctions; for example Parkinson's disease can be considered to be a secondary hyperglutamatergic disease. Additionally, glutamate is an essential factor in the plasticity response of the basal-ganglia. However, opposite to previous suggestions, the NMDA-receptor blocker MK-801 does not prevent psychostimulant- nor morphine-induced day to day increase (sensitization) of locomotion. Also the day to day increase of haloperidol-induced catalepsy was not prevented by MK-801. PMID- 9871436 TI - The role of NMDA receptors in the slow neuronal degeneration of Parkinson's disease. AB - Parkinson's disease is a disorder, in which neurons of various neuronal systems degenerate. Furthermore, in such degenerating neurons, the cytoskeleton seems to be affected. In this respect, Parkinson's disease resembles Alzheimer's disease. Since it has been shown, that elevated levels of intracellular calcium can disrupt the cytoskeleton and that the stimulation of glutamate (NMDA) receptors can cause high intracellular concentrations of calcium, it has been suggested, that the stimulation of glutamate receptors plays a role in the slow degeneration in Alzheimer's and Parkinson's disease. In case of the degeneration of the dopaminergic nigrostriatal system in Parkinson's disease, neurons that contain calcium binding protein appear to be less vulnerable than the neurons that lack it, suggesting that calcium binding protein might protect these neurons from degeneration by preventing that cytosolic calcium concentrations increase excessively. And, since there is in the nigrostriatal system a glutamatergic afferent pathway (the prefrontonigral projection) and since dopaminergic nigrostriatal neurons contain postsynaptic NMDA receptors, glutamatergic excitation may play a role in the degeneration of the nigrostriatal system in Parkinson's disease. If so, it may be possible to protect the neurodegeneration of these dopaminergic neurons by NMDA receptor antagonists. PMID- 9871435 TI - The role of striatal glutamate receptors in models of Parkinson's disease. AB - The aim of the study was to examine the effect of antagonists of the NMDA receptor on the parkinsonian-like muscle rigidity in rats. Reserpine and haloperidol increased the muscle resistance of the hind foot to passive movements, as well as the reflex electromyographic (EMG) activity in the gastroenemius and tibialis anterior muscles. MK-801 (0.32-1.28 mg/kg s.c.), an uncompetitive antagonist of the NMDA receptor, and L-701,324 (5-40 mg/kg i.p.), an antagonist of the glycine site, reduced the muscle tone and the reflex EMG activity enhanced by reserpine or haloperidol. AP-5 (2 and 5 micrograms/0.5 microliter), a competitive antagonist of the NMDA receptor, and 5,7 dichlorokynurenic acid (1.0-4.5 micrograms/0.5 microliter), the glycine site antagonist injected bilaterally into the rostral striatum, inhibited the muscle rigidity induced by haloperidol. In contrast, AP-5, injected alone bilaterally into the intermediate-caudal striatum induced muscle rigidity. The present results suggest that: (1) the inhibitory effect of the NMDA receptor antagonists on the parkinsonian-like muscle rigidity depends, at least partly, on their action on the rostral striatum; (2) the blockade of NMDA receptors in the intermediate-caudal striatum may reduce the beneficial impact of these compounds. PMID- 9871437 TI - Modulation of motor behaviour by NMDA- and cholecystokinin-antagonism. AB - Motor behaviour relies on complex neurochemical interactions in the basal ganglia, in particular the striatum. Antagonistic influences in this region are exerted by afferent projections from, on the one hand, the ventral mesencephalon, utilizing dopamine as a transmitter, and, on the other hand, from the cerebral cortex, signalling by the excitatory amino acid glutamate. The activity in both these neuronal populations appears to be regulated by the neuropeptide cholecystokinin. This article concentrates on interactions between cholecystokinin and glutamate, summarizing some recent morphological, biochemical and behavioural findings. It is suggested that cholecystokinin, acting via the cholecystokininB receptor, potentiates the glutamatergic excitatory input to the striatum. PMID- 9871438 TI - Adenosine A2A receptors inhibit the conductance of NMDA receptor channels in rat neostriatal neurons. AB - Whole-cell patch clamp experiments were carried out in rat striatal brain slices. In a subset of striatal neurons (70-80%), NMDA-induced inward currents were inhibited by the adenosine A2A receptor selective agonist CGS 21680. The non selective adenosine receptor antagonist 8-(p-sulphophenyl)-theophylline and the A2A receptor selective antagonist 8-(3-chlorostyryl)caffeine abolished the inhibitory action of CGS 21680. Intracellular GDP-beta-S, which is known to prevent G protein-mediated reactions, also eliminated the effect of CGS 21680. Extracellular dibutyryl cAMP, a membrane permeable analogue of cAMP, and intracellular Sp-cAMPS, an activator of cAMP-dependent protein kinases (PKA), both abolished the CGS 21680-induced inhibition. By contrast, Rp-cAMPS and PKI 14 24 amide, two inhibitors of PKA had no effect. Intracellular U-73122 (a phospholipase C inhibitor) and heparin (an inositoltriphosphate antagonist) prevented the effect of CGS 21680. Finally, a more efficient buffering of intracellular Ca2+ by a substitution of EGTA (11 mM) by BAPTA (5.5 mM) acted like U-73122 or heparin. Hence, A2A receptors appear to negatively modulate NMDA receptor channel conductance via the phospholipase C/inositoltriphosphate/Ca2+ pathway rather than the adenylate cyclase/PKA pathway. PMID- 9871439 TI - Antagonists of glutamate in the treatment of Parkinson's disease: from laboratory to the clinic. PMID- 9871440 TI - Effects of glutamate antagonists on the activity of aromatic L-amino acid decarboxylase. AB - This study examines the hypothesis that glutamate tonically suppresses the activity of the enzyme aromatic L-amino acid decarboxylase (AADC), and hence the biosynthesis of dopamine, to explain how antagonists of glutamate receptors might potentiale the motor actions of L-DOPA in animal models of Parkinson's disease. A variety of glutamate antagonists were therefore administered acutely to normal rats, which were sacrificed 30-60 min later and AADC activity assayed in the substantia nigra pars reticulata (SNr) and corpus striatum (CS). The NMDA receptor-ion channel antagonists MK 801, budipine, amantadine, memantine and dextromethorphan all caused a pronounced in creased in AADC activity, more especially in the SNr than CS. The NMDA glycine site antagonist (R)-HA 966 produced a modest increase in AADC activity in the CS but not SNr, whilst the NMDA polyamine site antagonist eliprodil, the NMDA competitive antagonist CGP 40116 and the AMPA antagonist NBQX were without effect. The results suggest that an increase in dopamine synthesis might contribute to the L-DOPA-facilitating actions of some glutamate antagonists. PMID- 9871441 TI - Role of excitatory amino acids in the ventral tegmental area for central actions of non-competitive NMDA-receptor antagonists and nicotine. AB - The putative role of non-NMDA excitatory amino acid (EAA) receptors in the ventral tegmental area (VTA) for the increase in dopamine (DA) release in the nucleus acumbens (NAC) and the behavioural stimulation induced by systemically administered dizocilpine (MK-801) was investigated. Microdialysis was utilized in rats with probes in the VTA and NAC. The VTA was perfused with the AMPA and kainate receptor antagonist CNQX (0.3 or 1.0 mM) or vehicle and dialysates from the NAC were analyzed with high-performance liquid chromatography for DA. Forty min after onset of CNQX or vehicle perfusion of the VTA MK-801 (0.1 mg/kg) was injected subcutaneously (s.c.). Subsequently, typical MK-801 induced behaviours were assessed. The MK-801 induced hyperlocomotion was associated with a 50% increase of DA levels in NAC dialysates. Both the MK-801 evoked hyperlocomotion and DA release in the NAC were effectively antagonized by CNQX perfusion of the VTA. However, by itself the CNQX or vehicle perfusion of the VTA did not affect DA levels in NAC or the rated behaviours. The results indicate that MK-801 induced hyperlocomotion and increased DA release in the NAC are largely elicited within the VTA via activation of non-NMDA EAA receptors, tentatively caused by locally increased EAA release. In contrast, the enhanced DA output in the NAC induced by systemic nicotine (0.5 mg/kg s.c.) was not antagonized by intra VTA infusion of CNQX (0.3 or 1.0 mM), but instead by infusion of the NMDA receptor antagonist AP-5 (0.3 or 1.0 mM) into the VTA, which by itself did not alter DA levels in the NAC. Thus, the probably indirect, EAA mediated activation of the mesolimbic DA neurons in the VTA by MK-801 and nicotine, respectively, seems to be mediated via different glutamate receptor subtypes. PMID- 9871442 TI - Role of excitatory amino acids in the regulation of dopamine synthesis and release in the neostriatum. AB - We have explored the role of excitatory amino acids in the increased dopamine (DA) release that occurs in the neostriatum during stress-induced behavioral activation. Studies were performed in awake, freely moving rats, using in vivo microdialysis. Extracellular DA was used as a measure of DA release; extracellular 3,4-dihydroxyphenylalanine (DOPA) after inhibition of DOPA decarboxylase provided a measure of apparent DA synthesis. Mild stress increased the synthesis and release of DA in striatum. DA synthesis and release also were enhanced by the intra-striatal infusion of N-methyl-D-aspartate (NMDA), an agonist at NMDA receptors, and kainic acid, an agonist at the DL-alpha-amino-3 hydroxy-5-methyl-4-isoxazole-4-propionate (AMPA)/kainate site. Stress-induced increase in DA synthesis was attenuated by co-infusion of 2-amino-5 phosphonovalerate (APV) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), antagonists of NMDA and AMPA/kainate receptors, respectively. In contrast, intrastriatal APV, CNQX, or kynurenic acid (a non-selective ionotropic glutamate receptor antagonist) did not block the stress-induced increase in DA release. Stress-induced increase in DA release was, however, blocked by administration of tetrodotoxin along the nigrostriatal DA projection. It also was attenuated when APV was infused into substantia nigra. Thus, glutamate may act via ionotropic receptors within striatum to regulate DA synthesis, whereas glutamate may influence DA release via an action on receptors in substantia nigra. However, our method for monitoring DA synthesis lowers extracellular DA and this may permit the appearance of an intra-striatal glutamatergic influence by reducing a local inhibitory influence of DA. If so, under conditions of low extracellular DA glutamate may influence DA release, as well as DA synthesis, by an intrastriatal action. Such conditions might occur during prolonged severe stress and/or DA neuron degeneration. These results may have implications for the impact of glutamate antagonists on the ability of patients with Parkinson's disease to tolerate stress. PMID- 9871443 TI - Direct and indirect presynaptic control of dopamine release by excitatory amino acids. AB - Dopamine (DA) release from nerve terminals of the nigrostriatal DA neurons not only depends on the activity of nigral DA cells but also on presynaptic regulation. Glutamatergic neurons of cortical origin play a prominent role in these presynaptic regulations. The direct glutamatergic presynaptic control of DA release is mediated by N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5 methyl-4-isoxazole-4-propionate (AMPA) receptors, located on DA nerve terminals. In addition, by acting on striatal target cells, these glutamatergic neurons contribute also to indirect regulations of DA release involving several transmitters such as GABA, acetylcholine and neuropeptides. Diffusible messengers such as nitric oxide (NO) or arachidonic acid (AA) which are particularly formed under the stimulation of NMDA receptors may also participate to the regulation of DA release. In the present study, it will be shown that the co-application of NMDA and carbachol synergistically increases the release of [3H]-DA and that this effect is reduced by mepacrine or 4-bromophenacylbromide (10(-7) M), two inhibitors of PLA2. Therefore endogenously released AA induced by the co stimulation of NMDA and cholinergic receptors seems to be involved, at least partly, in the release of DA. PMID- 9871444 TI - Role of glutamate in neurodegeneration of dopamine neurons in several animal models of parkinsonism. AB - Although controversial, studies with methamphetamine and MPTP suggest a link between glutamate-mediated excitotoxicity and degeneration of dopamine cells. Both compounds are thought to create a metabolic stress. To further explore glutamate actions in DA degeneration, we investigated the effects of other metabolic inhibitors. In mesencephalic cultures, DA cell loss produced by 3-NPA or malonate was potentiated by NMDA and prevented by MK-801. In vivo, striatal DA loss produced by intranigral infusions of malonate was also potentiated by intranigral NMDA and prevented by systemic MK-801. In contrast, systemic MK-801 did not prevent DA loss produced by intrastriatal malonate. Intrastriatal MK-801 or CGS 19755 did attenuate DA loss in METH-treated mice, but was confounded by the findings that METH-induced hyperthermia, an important component in toxicity, was also attenuated. Taken together, the data support the hypothesis of NMDA receptor involvement in degeneration of DA neurons. Furthermore, the data also suggest that this interaction is likely to occur in the substantia nigra rather than in the striatum. PMID- 9871446 TI - Neurobiology of the nitric oxide in the nervous system. Basic and clinical perspectives. PMID- 9871445 TI - Blockade of glutamatergic transmission as treatment for dyskinesias and motor fluctuations in Parkinson's disease. AB - In animal models of Parkinson's disease (PD), glutamate antagonists diminish levodopa (LD)-associated motor fluctuations and dyskinesias. We sought to investigate if these preclinical observations can be extended to the human disease, by evaluating the effects of three non-competitive NMDA antagonists (dextrorphan, dextromethorphan and amantadine) on the motor response to LD in patients with advanced PD. In four separate trials, adjuvant therapy with these drugs reduced LD-induced dyskinesias and motor fluctuations. These findings support the view that drugs acting to inhibit glutamatergic transmission at the NMDA receptor can ameliorate LD associated motor response complications. PMID- 9871447 TI - Nitric oxide as a peripheral and central mediator in temperature regulation. AB - In animals including humans nitric oxide (NO) serves as a biological messenger both peripherally at neuroeffector junctions and in the central nervous system where it modulates neuronal activity. Evidence for the involvement of NO in homeostatic control is accumulating also for temperature regulation in homeotherms. In the periphery an auxiliary role in the vasomotor control of convective heat transfer to heat dissipating surfaces and modulation of thermoregulatory heat generation, especially in brown adipose tissue as the site of nonshivering thermogenesis, are discussed as NO actions. At the central level a thermolytic role of NO in thermoregulation as well as in fever is assumed, however, experimental data opposing this view suggest that topical specificity may be important. At the level of single neurons, the observed interrelationships between thermosensitivity and responsiveness to NO are still not consistent enough to reconcile these data with the effects of NO-donors and inhibitors of NO synthase on temperature regulation. PMID- 9871448 TI - Nitric oxide in the pathophysiology of hyperthermic brain injury. Influence of a new anti-oxidant compound H-290/51. A pharmacological study using immunohistochemistry in the rat. AB - The possibility that nitric oxide (NO) is involved in the pathophysiology of brain injury caused by heat stress (HS) was examined using neuronal nitric oxide synthase (NOS) immunohistochemistry in a rat model. In addition, to find out a role of oxidative stress in NOS upregulation and cell injury, the effect of a new antioxidant compound H-290/51 (Astra Hassle, Molndal, Sweden) was examined in this model. Subjection of conscious young rats to 4 h HS in a biological oxygen demand (BOD) incubator at 38 degrees C resulted in a marked upregulation of NOS in many brain regions compared to control rats kept at room temperature (21 +/- 1 degree C). This NOS immunoreactivity was found mainly in distorted neurons located in the edematous regions not normally showing NOS activity. Breakdown of the blood-brain barrier (BBB) permeability, increase in brain water content and marked neuronal, glial and myelin reaction were common findings in several brain regions exhibiting upregulation of NOS activity. Pretreatment with H-290/51 significantly attenuated the upregulation of NOS in rats subjected to HS. In these animals breakdown of the BBB permeability, edema and cell changes were considerably reduced. Our results suggest that hyperthermic brain injury is associated with a marked upregulation of NOS activity in the CNS and this upregulation of NOS and concomitant cell injury can be reduced by prior treatment with an antioxidant compound H 290/51. These observations indicate that oxidative stress seems to be an important endogenous signals for NOS upregulation and cell reaction in hyperthermic brain injury. PMID- 9871449 TI - Spinal nerve lesion induces upregulation of neuronal nitric oxide synthase in the spinal cord. An immunohistochemical investigation in the rat. AB - The possibility nitric oxide (NO) is involved the neurodegenrative mechanisms in the spinal cord following a chronic peripheral nerve lesion was examined using NOS immunohistochemistry. Spinal nerve lesion at L-5 and L-6 level was produced according to the Chung model, a model of neuropathic pain and rats were allowed to survive for 8 weeks. In one group of animals L-NAME was given intraperitoneally (1-2 mg/kg, i.p. daily) for 6 weeks. Sham operated rats, in which the spinal nerve was exposed but not ligated, served as controls. Ligation of spinal nerves in rats resulted in an upregulation of NOS which was most pronounced in the ipsilateral gray matter of the spinal cord compared to the contralateral side. In these rats, ultrastructural investigations showed distorted neurons, membrane disruption and myelin vesiculation. Sham operated rats did not show either NOS upregulation or structural changes in the spinal cord. Pretreatment with L-NAME significantly reduced NOS upregulation and the structural changes in the spinal cord were less pronounced. These observations strongly indicate a putative role of NOS in the pathophysiology of chronic nerve lesion. Our results may provide a new strategy to treat chronic neuropathic pain or to minimise neurodegeneration in the patients suffering from such diseases of the nervous system. PMID- 9871450 TI - Effect of glutamate and angiotensin II on whole cell currents and release of nitric oxide in the rat subfornical organ. AB - Blood-borne angiotensin II (AngII) is known to mediate water-intake by its excitatory effect on neurons in the subfornical organ (SFO). Conversely, nitric oxide (NO) has exclusively inhibitory effects on rat SFO-neurons and on SFO mediated water-intake. Extracellular and patch-clamp recordings from freshly dissociated rat SFO-neurons showed that glutamate activates AngII-sensitive SFO neurons by opening ligand-gated cation channels. An immunocytochemical study showed that activation of glutamate receptors increased the concentration of the inhibitory second messenger cGMP in the SFO. A model is proposed suggesting that NO protects SFO-neurons from overexcitability by excitatory neurotransmitters. PMID- 9871451 TI - Brain derived neurotrophic factor and insulin like growth factor-1 attenuate upregulation of nitric oxide synthase and cell injury following trauma to the spinal cord. An immunohistochemical study in the rat. AB - The possibility that brain derived neurotrophic factor (BDNF) and insulin like growth factor-1 (IGF) induced neuroprotection is influenced by mechanisms involving nitric oxide was examined in a rat model of focal spinal cord injury. BDNF or IGF-I (0.1 microgram/10 microliters in phosphate buffer saline) was applied topically 30 min before injury on the exposed spinal cord followed by repeated doses of growth factors immediately before and 30 min after injury. Thereafter application of BDNF or IGF was carried out at every 1 h interval until sacrifice. Five hours after injury, the tissue pieces from the T9 segment were processed for nNOS immunostaining, edema and cell injury. Untreated injured rats showed a profound upregulation of nNOS which was most pronounced in the nerve cells of the ipsilateral side. A marked increase in the blood-spinal cord barrier (BSCB) permeability to 125I-albumin, water content and cell injury in these perifocal segments was also found. Pretreatment with BDNF and IGF significantly reduced the upregulation of nNOS in the spinal cord. This effect of the growth factors was most pronounced in the contralateral side. Rats treated with these neurotrophic factors showed much less signs of BSCB damage, edema and cell injury. These results suggest that BDNF and IGF pretreatment is neuroprotective in spinal cord injury and that these neurotrophic factors have the capacity to down regulate nNOS expression following trauma to the spinal cord. Our data provide new experimental evidences which suggest that BDNF and IGF may exert their potential neuroprotective effects probably via regulation of NOS activity. PMID- 9871452 TI - Spinal cord evoked potentials and edema in the pathophysiology of rat spinal cord injury. Involvement of nitric oxide. AB - The possibility that nitric oxide is somehow involved in the early bioelectrical disturbances following spinal cord injury in relation to the later pathophysiology of the spinal cord was examined in a rat model of spinal cord trauma. A focal trauma to the rat spinal cord was produced by an incision of the right dorsal horn of the T 10-11 segments under urethane anaesthesia. The spinal cord evoked potentials (SCEP) were recorded using epidural electrodes placed over the T9 and T12 segments of the cord following supramaximal stimulation of the right tibial and sural nerves in the hind leg. Trauma to the spinal cord significantly attenuated the SCEP amplitude (about 60%) immediately after injury which persisted up to 1 h. However, a significant increase in SCEP latency was seen at the end of 5 h after trauma. These spinal cord segments exhibited profound upregulation of neuronal nitric oxide synthase (NOS) immunoreactivity, and the development of edema and cell injury. Pretreatment with a serotonin synthesis inhibitor drug p-chlorophenylalanine (p-CPA) or an anxiolytic drug diazepam significantly attenuated the decrease in SCEP amplitude, upregulation of NOS, edema and cell injury. On the other hand, no significant reduction in SCEP amplitude, NOS immunolabelling, edema or cell changes were seen after injury in rats pretreated with L-NAME. These observations suggest that nitric oxide is somehow involved in the early disturbances of SCEP and contribute to the later pathophysiology of spinal cord injury. PMID- 9871453 TI - Voltage-dependent sodium channels and excitatory amino acids. PMID- 9871454 TI - Glutamate release inhibitors: a critical assessment of their action mechanism. AB - A number of important experimental data do not support the widespread hypothesis that Na(+)-channels block is cerebroprotective, essentially because it reduces presynaptic glutamate release: (i) the inhibition of exocytosis by these compounds is not specific to glutamate; (ii) aspartate efflux produced by various stimuli was also reduced, but aspartate cannot be released by exocytosis because it is not concentrated within presynaptic vesicles; and (iii) glutamate accumulated extracellularly during ischaemic or traumatic insult to the CNS is mainly of cytosolic origin. As an alternative, we propose that use-dependent Na(+)-channel blockers enhance the resistance of nerve cells to insults, primarily by decreasing their energy demand, and that reduced efflux of glutamate and other compounds is a consequence of attenuated cellular stress. PMID- 9871455 TI - Neuroprotection--rationale for pharmacological modulation of Na(+)-channels. AB - The primary factor detrimental to neurons in neurological disorders associated with deficient oxygen supply or mitochondrial dysfunction is insufficient ATP production relative to their requirement. As a large part of the energy consumed by brain cells is used for maintenance of the Na+ gradient across the cellular membrane, reduction of energy demand by down-modulation of voltage-gated Na(+) channels is a rational strategy for neuroprotection. In addition, preservation of the inward Na+ gradient may be beneficial because it is an essential driving force for vital ion exchanges and transport mechanisms such as Ca2+ homeostasis and neurotransmitter uptake. PMID- 9871456 TI - The importance of voltage-dependent sodium channels in cerebral ischaemia. AB - Strategies for the treatment of thromboembolic stroke are based on restoring the blood flow as soon as possible and protecting the neurons from the deleterious consequences of cerebral ischaemia. Interest has focused on blockers of voltage dependent Na+ channels as potential neuroprotective agents because they prevent neuronal death in various experimental models of cerebral ischaemia and act cytoprotectively in models of white matter damage. Although several Na+ blockers are currently being tested in various phases of clinical development, most of these agents are relatively weak and unspecific. I therefore consider it worthwhile to search for molecules which specifically block voltage-dependent Na+ channels for the treatment of cerebral ischaemia. PMID- 9871457 TI - Observations relative to the neurotoxicity and neurotoxic potential of amino acids. PMID- 9871458 TI - Destruction of catecholamine-containing neurons by 6-hydroxydopa, an endogenous amine oxidase cofactor. AB - The amino acid, 6-hydroxydopa (6-OHDOPA), found at the active site of amine oxidases, exists as a keto-enol. Exogenously administered 6-OHDOPA is an excitotoxin like beta-N-oxalylamino-L-alanine (BOAA) and beta-N-methylamino-L alanine (BMAA), acting at the non-N-methyl-D-aspartate (non-NMDA) alpha-amino-3 hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptor. BMAA and BOAA are causal factors of neurolathyrism in humans. Much exogenously administered 6 OHDOPA is biotransformed by aminoacid decarboxylase (AADC) to the highly potent and catecholamine-(CA) selective neurotoxin, 6-hydroxydopamine (6-OHDA). 6-OHDOPA destroys locus coeruleus noradrenergic perikarya and produces associated denervation of brain by norepinephrine-(NE) containing fibers. Opiopeptides and opioids enhance neurotoxic effects of 6-OHDOPA on noradrenergic nerves, by a naloxone-reversible process. An understanding of mechanisms underlying neurotoxic effects of 6-OHDOPA can be helpful in defining actions of known and newfound amino acids and for investigating their potential neurotoxic properties. PMID- 9871459 TI - Molecular mechanisms of cellular injury produced by neurotoxic amino acids that generate reactive oxygen species. AB - There is now strong experimental evidence that the basic precursors for the synthesis of catechol(amine) and indolamine neurotransmitters, tyrosine and tryptophan can act as generators of ROS (reactive oxygen species): peroxides, superoxide and peroxyradicals. The consequences of free radicals formation from precursors during oxidative degradation process, their possible participation in electron transfer/addition reactions and chain processes involving cell antioxidant defense system were presented and discussed. Although the generation of neurotoxic ROS by tyrosine and tryptophan is accepted to occur in the presented model systems, doubts can exist as to the situation in vivo, which may be completely different and remain to be explored. The relevance of the present findings with regard to a variety of neurological diseases cannot be ignored. PMID- 9871460 TI - Pharmacological approaches to counter the toxicity of Dopa. AB - Dopa and related catecholamines and their degradation products have been demonstrated to have neurotoxic potential in a number of cellular and in vivo experiments. Several mechanisms have been hypothesized to be involved including generation of prooxidant products that subsequently oxidize membrane lipids and exposed macromolecules. We have utilized a neuronal culture of cerebellar granule cells to study the toxicity of Dopa and the ability of various neuroprotective and antiparkinsonian compounds to offer protection therefrom. This model is apparently based on the ability of Dopa to non-enzymatically induce an oxidative injury to the neuronal cultures. Evidence for this arises from the equal neurotoxic potency of L- and D-Dopa in these cells and the ability of catalase, superoxide dismutase and glutathione to protect the neurons from this toxicity. Further, we found that the neuroprotective antioxidant, PNU-101033 is more effective and potent than vitamin E and deprenyl in this regard. Similarly the D2/D3 agonist, pramipexole is also capable of blocking Dopa toxicity in this model and this effect is independent of dopamine receptor affinity as both enantiomers are equally potent in this assay but disparate in receptor affinity. Also the protection by pramipexole is accompanied by the preservation of reduced glutathione. Thus, this activity seems to be a function of the oxidation potential of pramipexole and it's consequent antioxidant property. Potent antioxidants are effective blockers of Dopa toxicity. If the mechanisms involved in this toxicity have relevance to the progression of Parkinson's pathology in Dopa treated (or untreated) patients, these compounds have the potential to alter the course of the illness. PMID- 9871461 TI - Release of endogenous excitatory amino acids in the neostriatum of the rat under physiological and pharmacologically-induced conditions. AB - There is immunohistochemical evidence suggesting that glutamate (Glu) is released from nerve terminals and acts, via several receptor subtypes, as a major excitatory neurotransmitter in the cortico-striatal pathway of the rat. Aspartate (Asp) is also present in cortico-striatal neurons, but its role as a neurotransmitter has been questioned, since, in contrast to Glu, it has not been demonstrated in presynaptic vesicles. Glu and Asp can be found at submicroM concentrations in the extracellular compartment of most areas of the basal ganglia. Their concentrations are largely regulated by transport mechanisms, but also by a synaptotagmin-dependent exocytotic release, and are sufficiently high to occupy junctional and extrajunctional receptors. We have investigated whether Glu and Asp release in the neostriatum can be selectively modulated by different neuronal systems. Dopamine (DA) and cholecystokinin (CCK) selectively stimulate Asp release, via D1 and CCKB receptor subtypes, respectively. Also opioid kappa agonists increase Asp release. We propose that the selective modulation of Asp release by D1-, CCKB- and kappa-agonists involves striatal neurons containing Asp, but not Glu. In contrast, local perfusion with the mu-opioid antagonist D Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP) increases both Glu and Asp release. This effect is probably exerted on cortico-striatal terminals, via presynaptic inhibitory mu-receptors. Thus, these results demonstrate that extracellular levels of Glu and Asp are modulated differentially by different neuronal systems, and suggest that in the neostriatum of the rat there are neuronal populations using Glu and/or Asp as messenger(s). PMID- 9871462 TI - GlycineB recognition site of NMDA receptors and its antagonists. PMID- 9871463 TI - Modulation of NMDA receptors by glycine--introduction to some basic aspects and recent developments. AB - Glycine is a co-agonist at NMDA receptors and it's presence is a prerequisite for channel activation by glutamate or NMDA. Physiological concentrations reduce one form of NMDA receptor-desensitization. Interactions between the glycineB site and other domains of the NMDA receptor are complex and include the glutamate, Mg2+ and polyamines sites. Glycine shows different affinities at various NMDA receptor subtypes probably via to allosteric interactions between NMDA2 subunits and the glycine recognition site on the NMDAR1 subunit. There is still some debate whether the glycineB site is saturated in vivo but it seems likely that this depends on regional differences in receptor subtype expression, local glycine or D-serine concentrations and the expression of specific glycine transporters. GlycineB antagonists and partial agonists have been reported to have good therapeutic indices as neuroprotective agents against focal ischaemia and trauma, anti-epileptics, anxiolytics, anti-psychotomimetics and in models of chronic pain. They clearly lack two potentially serious side effects classically associated with NMDA receptor blockade, namely neurodegenerative changes in the cingulate/retrosplenial cortex and psychotomimetic-like effects. This improved therapeutic profile may be partially due to the ability of full glycineB antagonists to reveal glycine-sensitive desensitization and possibly also via functional and/or regional NMDA receptor subtype selectivity. PMID- 9871464 TI - Effects of NMDA receptor antagonists on nociceptive responses in vivo: comparison of antagonists acting at the glycine site with uncompetitive antagonists. AB - We have shown that members of a new series of tricyclic pyridophthalazine diones, defined as glycineB site NMDA antagonists in vitro, are selective and systemically active NMDA antagonists in vivo. In electrophysiological tests in alpha-chloralose anaesthetised rats, these compounds reduced nociceptive reflex responses. In conscious rats they displayed analgesic properties. These glycineB antagonists were compared electrophysiologically with several uncompetitive NMDA channel blockers. The degree of voltage dependence previously reported in vitro related to the effectiveness of the agents against different amplitude nociceptive responses of spinal cord neurones in vivo. PMID- 9871465 TI - Novel glycineB antagonists show neuroprotective activity in vivo. AB - The degeneration or dysfunction of cholinergic neurons within the basal forebrain of patients with Alzheimer's disease (AD) may be related to the vulnerability of these cells to endogenous glutamate (Beal, 1995; Greenamyre and Young, 1989). The administration of drugs that attenuate the toxic actions of glutamate in the early stages of the disease might significantly delay its rate of progression. Two approaches to neuroprotection from endogenous glutamatergic function were investigated and found to be effective: blockade of voltage-dependent, NMDA-type glutamate receptor channels and antagonism of an NMDA-receptor related glycineB modulatory site. PMID- 9871466 TI - Ligands of the NMDA receptor-associated glycine recognition site and motor behavior. AB - Motor behavior critically depends on glutamatergic functions in the basal ganglia (BG). The dorsal and ventral striatum--the main input structures of the BG--are involved in modulation of stereotyped sniffing behavior, locomotion, catalepsy and prepulse inhibition. The effects of the NMDA receptor have been well characterized in respect to motor behavior in the past. The function of the allosteric glycine site was however disregarded until now, because brain penetrating ligands were missing. The present study summarized the motor behavioral profile of several glycine site ligands (7-chlorokynurenate, ACEA 1021, MRZ-2/576, (+) HA-966, D-cycloserine and felbamate). It is shown that through blockade of the glycine site of the NMDA receptor a distinct behavioral profile can be obtained. PMID- 9871467 TI - GlycineB antagonists as potential therapeutic agents. Previous hopes and present reality. AB - It is not clear what therapeutic application is most likely for agents blocking glycine site of the NMDA receptors (glycineB). Majority of the studies to date used either glycineB antagonists with doubtful brain penetration or partial agonists. Following systemic administration to rats of our newly developed glycineB antagonists (MRZ 2/570; 2/571 and 2/576) and L-701,324 (MSD) as a reference agent the following behavioural effects were observed: weak (if any) antiparkinsonian-like effects, lack of anxiolytic activity, inhibition of physical and motivational aspects of morphine dependence and neuroprotective activity in global ischaemia. The side effects include: sedation, ataxia, and myorelaxation. We detected neither vacuolisation in the cingulate cortex nor impairment of pre-pulse inhibition indicating lack of psychotomimetic potential. PMID- 9871468 TI - Neuroactive tryptophan metabolites: focus on kynurenines. PMID- 9871469 TI - Regulation of kynurenic acid levels in the developing rat brain. AB - Several brain-specific mechanisms control the formation of the endogenous excitatory amino acid receptor antagonist kynurenic acid (KYNA) in the adult rat brain. Two of these, dopaminergic neurotransmission and cellular energy metabolism, were examined in the brain of immature (postnatal day 7) rats. The results indicate that during the early postnatal period cerebral KYNA synthesis is exceptionally amenable to modulation by dopaminergic mechanisms but rather insensitive to fluctuations in cellular energy status. These findings may be of relevance for the role of KYNA in the function and dysfunction of the developing brain. PMID- 9871470 TI - Expression of the kynurenine enzymes in macrophages and microglial cells: regulation by immune modulators. AB - The regulation of the expression of indoleamine 2,3-dioxygenase (IDO) was studied in cloned murine macrophages (MT2) and microglial (N11) cells. Both cell lines express IDO and inducible nitric oxide synthase activity after interferon-gamma (IFN-gamma) stimulation. The regulation of IDO expression appears to differ in the two cell lines. Nitric oxide (NO) production negatively modulates the expression of IDO activity in IFN-gamma-primed macrophages, thereby indicating a cross-talk between the kynurenine and nitridergic pathways in these cells. Conversely, this down-regulation of IDO activity by NO does not occour in microglial cells. A differential regulation of IDO expression in the two cell lines was also observed with LPS and picolinic acid. Together with previous findings, these results indicate the existence of marked differences in the regulation of the expression of the kynurenine pathway enzymes between macrophages and microglial cells. PMID- 9871471 TI - Role of zinc in blockade of excitotoxic action of quinolinic acid by picolinic acid. AB - This study examined whether picolinic acid (PIC) inhibits quinolinic acid (QUIN) induced excitotoxicity through zinc chelation. Injection of QUIN into the nucleus basalis magnocellularis significantly depleted cortical choline acetyltransferase activity 7 days post injection and PIC inhibited this response. Zinc augmented the QUIN- but not NMDA-induced response. When PIC was co-administered with zinc, PIC failed to attenuate the QUIN-induced response. The inhibition of QUIN-induced cholinergic toxicity by PIC may involve chelation of zinc. PMID- 9871472 TI - Effects of the 3-hydroxyanthranilic acid analogue NCR-631 on anoxia-, IL-1 beta- and LPS-induced hippocampal pyramidal cell loss in vitro. AB - The kynurenine pathway intermediate 3-hydroxyanthranilic acid (3-HANA) is converted by 3-HANA 3,4-dioxygenase (3-HAO) to the putative neuropathogen quinolinic acid (QUIN). In the present study, the neuroprotective effects of the 3-HANA analogue and 3-HAO inhibitor NCR-631 was investigated using organotypic cultures of rat hippocampus. An anoxic lesion was induced by exposing the cultures to 100% N2 for 150 min, resulting in a pronounced loss of pyramidal neurons, as identified using NMDA-R1 receptor subunit immunohistochemistry. NCR 631 provided a concentration-dependent protective effect against the anoxia. NCR 631 was also found to counteract the loss of pyramidal neurons in two models of neuroinflammatory-related damage; incubation with either LPS (10 ng/ml) or IL-1 beta (10 IU/ml). The findings suggest that NCR-631 has neuroprotective properties and that it may be a useful tool to study the role of kynurenines in neurodegeneration. PMID- 9871473 TI - A review of physiological and metabolic effects of essential amino acids. AB - The authors review ten essential amino acids with regard to their metabolic, physiologic and therapeutic effects throughout the human body. Physical properties of these biologically active compounds are discussed as a foundation for their diverse roles in special nitrogen containing products, neurotransmitters, and as alternative energy sources. Both normal and abnormal amino acid metabolism are considered in the areas of digestion, elimination of metabolic products, metabolic intermediates, and defects in these systems. Recent developments in therapeutic applications are further examined for clinical utility and as an economical alternative to traditional clinical treatment modalities. PMID- 9871474 TI - Modified chemotactic peptides: synthesis and activity of an azaTic-containing fMLP-OMe analogue. AB - The synthesis and the biological activity of a pseudopeptide analogue of the chemotactic N-formyltripeptide fMLP-OMe, containing the azaTic (3,4-dihydro-2(1H) phthalazinecarboxylic acid) residue replacing the native phenylalanine, is described. Whereas pseudopeptides containing linear alpha-azaamino acids are currently studied, data on the new group of analogues containing cyclic alpha-aza residues capable of limiting the rotameric distribution of the side chains (topological control) are just emerging in the literature. At our best knowledge, the here described [azaTic3]fMLP-OMe represents the first example of the introduction of this new type of alpha-aza residue into a natural bioactive peptide. PMID- 9871475 TI - Asymmetric imine-ene reactions: diastereofacial selective reactions with chiral glyoxylate-derived alpha-imino esters and asymmetric catalysis of enantiofacial selective reactions with prochiral alpha-imino esters. AB - The diastereofacial selective "imine-ene" reactions with alpha-imino esters, prepared from (-)-8-phenylmenthyl glyoxylate, are shown to provide an efficient entry to the asymmetric synthesis of alpha-amino acids. The feasibility study of the asymmetric catalysis is also reported on the enantiofacial selective ene reactions with prochiral alpha-imino esters. PMID- 9871476 TI - Microdialysis of excitatory amino acids in the periaqueductal gray of the rat after unilateral peripheral inflammation. AB - This study measured the release of glutamate (Glu) and aspartate (Asp) amino acid transmitters in the ventrocaudal compartment of the rat periaqueductal gray (PAG) following exposure to unilateral peripheral inflammation. The release of endogenous Glu and Asp from the rat ventrocaudal PAG was monitored with the microdialysis technique in unanesthetized, unrestrained rats. There was significant increase (1,300%) in the basal concentrations of Glu release in the 7 days Complete Freund's Adjuvant (CFA) treated group compared to 24 h mineral oil control group. Amino acid release was induced by infusing veratridine (75 microM, a sodium channel activator) directly through the 1 mm long dialysis probe. Perfusion of veratridine into the ventrocaudal PAG resulted in significant elevation of Glu and Asp amino acids. In the 24 h and 7 days CFA treated rats, veratridine-evoked release of Glu was significantly decreased in the lateral ventrocaudal PAG compared to control rats injected with mineral oil (CFA vehicle). The peak minus baseline concentrations of Glu in 24 h and 7 days CFA treated groups decreased 55.7% and 43.9%, respectively. In contrast, The basal and the peak minus baseline concentrations of Asp showed no significant change between control group and 24 h and 7 days CFA treated animals. The results provide direct evidence that Glu excitatory amino acid may be involved in nociception/nociception modulation pathway in the ventrocaudal PAG. PMID- 9871477 TI - Design and synthesis of small semi-mimetic peptides with immunomodulatory activity based on myelin basic protein (MBP). AB - Experimental allergic encephalomyelitis (EAE) is induced in susceptible animals by immunodominant determinants of myelin basic protein (MBP). Analogs of these disease-associated peptides have been identified with disease progression upon coimmunization. Usage of peptides, with disease-specific immunomodulatory capacity in vivo is limited, however, due to their sensitivity to proteolytic enzymes. Alternative approaches include the development of mimetic molecules which maintain the biological function of an original peptide, yet are stable and able to elicit their response in pharmacological quantities. A novel technique was employed to design a series of semi-mimetic peptides, based on the guinea pig MBP72-85 peptide used to induce EAE in Lewis rats. We used isonipecotic (iNip) and aminocaproic (Acp) acids as templates. Acp-MBP72-85 peptide derived analogues were effective in inducing EAE compared to iNip-peptide analogues which were ineffective at 350 micrograms. These findings suggest that the design and synthesis of semi-mimetic peptide molecules with immunomodulatory potential is possible and that eventually these molecules may form the basis for the development of novel and more effective disease-specific therapeutic agents. PMID- 9871478 TI - Analysis of HIV by entropy evolution rate. AB - We analyze the variation of HIV after infection by means of an information measure, called the entropy evolution rate. In our analysis, we use a part of the external glycoprotein gp120 including the V3 region observed from six patients. Then we could make the following two aspects clear; (1) the relation between the change of the entropy evolution rate and the appearance of symptoms of disease, and (2) the relation between the change of the entropy evolution rate and that of the CD4 count of the patients. PMID- 9871479 TI - Occurrence of free D-aspartate and aspartate racemase in the blood shell Scapharca broughtonii. AB - Substantial concentrations of D-aspartate were found in several tissues of Scapharca broughtonii together with approximately equal concentrations of L aspartate. The foot and mantle extracts also contained an aspartate racemase activity. The formation of L-aspartate from the D-enantiomer by the foot extract was apparently slower than the reverse reaction, and this unbalance seemed to be due to the presence of an enzyme activity which rapidly converted L-aspartate to L-alanine. The possible role of D-aspartate in the anaerobiosis was discussed. PMID- 9871480 TI - Free carnitine and acetyl carnitine plasma levels and their relationship with body muscular mass in athletes. AB - The purpose of the present study was to investigate the relationship between plasma carnitine concentration and body composition variation in relation to muscular and fat masses since there is no experimentally proved correlation between plasma carnitine and body masses. We used bioelectric impedance analysis (BIA), to determine body composition and to have a complete physical fitness evaluation. The post-absorptive plasma free carnitine and acetyl carnitine plasma levels, body composition as Fat-Free Mass (FFM) and Fat Mass (FM) in kg, as well as in percent of body mass, were analysed in 33 healthy subjects. A significant negative correlation was found between plasma acetyl carnitine and FFM in weight (kg) as well as in percent of body mass (respectively p < 0.0001; p < 0.01); a significant positive correlation was found only between FM in percent and plasma acetyl carnitine (p < 0.01). The observed negative correlation between plasma acetyl carnitine and muscular mass variation might reflect an oxidative metabolic muscle improvement in relation to muscular fat free mass increment and might be evidence that muscle metabolism change is in relation to plasma acetyl carnitine concentration. PMID- 9871481 TI - Renal excretion of gamma-carboxyglutamic acid and metabolic rate in 3-18 years old humans. AB - The modified amino acid gamma-carboxyglutamic acid (Gla) occurs in several proteins such as prothrombin, blood coagulation factors VII, IX and X, proteins C, S and Z as well as matrix Gla protein and osteocalcin. The amount of Gla excreted in urine is a common indicator of the whole-body degradation of these proteins. We have determined the renal excretion rates of Gla in 3, 6, 10, 14 and 18 years old male and female human subjects (n = 14 per age group and sex) and calculated the respective resting metabolic rates (RMR) on the basis of the body weights using published formulas. We found high correlations between the excretion rates of Gla (mumol/d/kg body weight) and the RMR (kJ/d/kg body weight) in the females (n = 70) of r = 0.70 (y = 0.003x + 0.29) and in the males (n = 70) of r = 0.70 (y = 0.0038x + 0.27) and in all subjects (n = 140) of r = 0.69 (y = 0.0035x + 0.27); p < 0.01. We postulate that in children and adolescents a causal relationship exists between the whole-body degradation rate of Gla containing proteins and the metabolic rate. PMID- 9871482 TI - System B0,(+)-mediated regulation of lysine transport in Caco-2 human intestinal cells. AB - We investigated whether lysine transport would be subject to adaptive regulation in Caco-2 human intestinal cells. The activity of Lys transport in Caco-2 cells decreased with increasing incubation time with 10 mM Lys. Among the two systems involved in Lys transport, the system b0,+ component was greatly decreased by incubating cells with 10 mM Lys, whereas the system y+ component did not change. These results suggest that system b0,+ mainly contributes to the adaptive regulation of Lys transport in Caco-2 cells. PMID- 9871483 TI - Changes in mediobasal hypothalamic dopamine and GABA release: a possible mechanism underlying taurine-induced prolactin secretion. AB - Taurine (Tau), a putative inhibitory amino acid neurotransmitter, has been shown to stimulate prolactin (PRL) release. Using ovariectomized, estrogen-replaced adult rats we investigated initially the effect of this amino acid, injected by different routes, on PRL secretion in vivo. Tau (100-500 mg/kg) had no effect on PRL release when given i.p.; 15 min after i.c.v. injection of Tau (3 mumoles), a significant increase in serum PRL levels was observed (78 +/- 9 ng/ml over basal levels, p < 0.01 vs. controls). In vitro (cultured anterior pituitary cells) PRL release was not affected by a 5 h incubation with Tau (10(-3)-10(-8) M). Basal dopamine (DA) or gamma-aminobutyric acid (GABA) output from superfused mediobasal hypothalamic fragments (MBH) was not affected by Tau (10(-3) M or 10(-5) M). However, during stimulation with KCl (50 mM), Tau (10(-3) M) significantly lowered DA release, and increased GABA output. It is concluded that Tau acts at a central level to increase PRL secretion, most probably by modulating the hypothalamic release of neurotransmitters controlling lactotroph function. PMID- 9871485 TI - Precursors of taurine in female genital tract: effects on developmental capacity of bovine embryo produced in vitro. AB - Two precursors of taurine have been studied: cysteamine and hypotaurine. Cysteamine has been quantified in genital secretions and found in follicular fluids of all species tested. On the contrary cysteamine was not detected (or traces) in tubal fluids of the same species. Addition of 50, 100 or 250 microM of cysteamine to the maturation medium used in the culturing of bovine oocytes did not improve the cleavage rate nor the embryo's developmental potential in vitro. Furthermore, at 250 microM, cysteamine seems to be toxic to the embryo. Addition of 0.5-1 mM hypotaurine to the bovine embryo culture medium improved significantly blastocyst production and quality. The respective roles of these 2 taurine precursors on maturation and embryo development are discussed. PMID- 9871484 TI - The effects of the beta 2-agonist drug clenbuterol on taurine levels in heart and other tissues in the rat. AB - The administration of a single subcutaneous dose of clenbuterol to rats altered the level of taurine in certain tissues. Taurine levels in cardiac tissue were significantly decreased 3 h after the administration of 250 micrograms/kg of clenbuterol and remained significantly depressed at 12 h post-dose only returning to control values by 24 h. The level of taurine in the liver increased 3 h after clenbuterol administration but was lower than the control value at 24 h post dose. Lung taurine levels were significantly lower than the control value at 12 hr post dose and remained depressed until 24 h post dose. Clenbuterol caused a significant increase in taurine levels in serum and muscle at 3 and 6 hr postdosing respectively but not at other time points. Serum creatine kinase (CK), activity was slightly but significantly raised at the 12 and 24 h time point. The effects of clenbuterol on tissue taurine content were not dose-dependent over the range studied (63-500 micrograms/kg). However taurine levels in the lung were significantly reduced at all doses and in the heart were significantly lower in the treated groups at all except the lowest dose, 12 h post dosing. Liver taurine levels were significantly increased at the highest dose of 500 micrograms/kg. The reduction of taurine concentrations in the heart, caused by clenbuterol, is of concern as taurine has been shown to have protective properties in many tissues especially the heart. PMID- 9871486 TI - Activation of adenosine A2 receptors enhances high K(+)-evoked taurine release from rat hippocampus: a microdialysis study. AB - The present study was designed to examine which type of adenosine receptors was involved in enhancement of high K(+)-evoked taurine release from in vivo rat hippocampus using microdialysis. Perfusion with 0.5 or 5.0 mM adenosine enhanced high K(+)-evoked taurine release. Perfusion with 2 microM R(-)-N6-2 phenylisopropyladenosine (PIA), a selective adenosine A1 receptor agonist, did not modulate taurine release. Perfusion with 1 microM 1,3-dipropyl-8 cyclopentylxanthine (DPCPX), a selective adenosine A1 receptor antagonist, increased taurine release. On the other hand, perfusion with 20 microM 2-[4-(2 carboxyethyl)phenethylamino]-5'-N-ethyl-carboxamide-adenos ine (CGS21680), a selective adenosine A2A receptor agonist, enhanced taurine release, while perfusion with 1 mM 3,7-dimethyl-propagylxanthine (DMPX), an adenosine A2 receptor antagonist, did not affect taurine release. These results demonstrate that adenosine enhances high K(+)-evoked taurine release via activation of adenosine A2A receptors from both neurons and glial cells of in vivo rat hippocampus. PMID- 9871487 TI - Taurine: protective properties against ethanol-induced hepatic steatosis and lipid peroxidation during chronic ethanol consumption in rats. AB - Alcohol was administered chronically to female Sprague Dawley rats in a nutritionally adequate totally liquid diet for 28 days. This resulted in hepatic steatosis and lipid peroxidation. Taurine, when co-administered with alcohol, reduced the hepatic steatosis and completely prevented lipid peroxidation. The protective properties of taurine in preventing fatty liver were also demonstrated histologically. Although alcohol was found not to affect the urinary excretion of taurine (a non-invasive marker of liver damage), levels of serum and liver taurine were markedly raised in animals receiving alcohol + taurine compared to animals given taurine alone. The ethanol-inducible form of cytochrome P-450 (CYP2E1) was significantly induced by alcohol; the activity was significantly lower than controls and barely detectable in animals fed the liquid alcohol diet containing taurine. In addition, alcohol significantly increased homocysteine excretion into urine throughout the 28 day period of ethanol administration; however, taurine did not prevent this increase. There was evidence of slight cholestasis in animals treated with alcohol and alcohol + taurine, as indicated by raised serum bile acids and alkaline phosphatase (ALP). The protective effects of taurine were attributed to the potential of bile acids, especially taurine conjugated bile acids (taurocholic acid) to inhibit the activity of some microsomal enzymes (CYP2E1). These in vivo findings demonstrate for the first time that hepatic steatosis and lipid peroxidation, occurring as a result of chronic alcohol consumption, can be ameliorated by administration of taurine to rats. PMID- 9871488 TI - Effects of taurine depletion on cell migration and NCAM expression in cultures of dissociated mouse cerebellum and N2A cells. AB - Cultures of dissociated cerebellum from 5- to 6-day-old mice as well as of the N2A neuronal cell line were exposed to guanidino ethane sulfonate (GES, 2-5 mM) to reduce the cellular taurine content. Control cultures were kept in culture medium or medium containing 2-5 mM GES plus 2-5 mM taurine to restore the intracellular taurine content. Taurine depletion led to changes in the expression of certain splice variants of NCAM mRNA such as the AAG and the VASE containing forms, while no differences were seen in the expression of the three forms of NCAM protein. In the N2A cells taurine depletion led to a decreased migration rate of the cells. The results suggest that the reduced migration rate of neurons caused by taurine depletion may be correlated to changes in expression of certain adhesion molecules such as NCAM. Moreover, taurine appears to be involved in regulation of transcription processes. PMID- 9871489 TI - Antidiuretic hormone infusion reduces taurine and NaCl-induced hypernatremia in the rat. AB - Rats drinking taurine and hypertonic saline (T + S) develop severe hypernatremia, but rats drinking either T or S alone do not. One hypothesis for this disruption of homeostasis is that the T + S combination interferes with the actions of antidiuretic hormone (ADH). Rats drinking T + S developed severe hypernatremia (170 mmol/L) by day 8 when infused with distilled water by osmotic minipumps, but maintained plasma sodium below 150 mmol/L when infused with ADH. Cumulative water balance in T + S drinkers receiving ADH was consistently higher than in those not receiving ADH. However the ratio of cumulative sodium balance to cumulative water balance suggests little uniform advantage to rats receiving ADH nor does comparison of urine osmolality in the two groups. Precisely how ADH administration reduces hypernatremia in T + S drinking rats remains unclear, but the hypothesis that T + S interferes with the action of ADH in its regulation of extracellular fluid volume and osmolality remains viable. PMID- 9871490 TI - Pharmacological characterization of the effects of taurine on calcium uptake in the rat retina. AB - Taurine is known to increase ATP-dependent calcium ion (Ca2+) uptake in retinal membrane preparations and in isolated rod outer segments (ROS) under low calcium conditions (10 microM) (Pasantes-Morales and Ordonez, 1982; Lombardini, 1991). In this report, ATP-dependent Ca2+ uptake in retinal membrane preparations was found to be inhibited by 5 microM cadmium (Cd2+), suggesting the involvement of cation channel activation. The activation of cGMP-gated cation channels, which are found in the ROS, is a crucial step in the phototransduction process. An inhibitor of cGMP-gated channels, LY83583, was found to inhibit taurine-stimulated ATP dependent Ca2+ uptake but had no effect on ATP-dependent Ca2+ uptake in the absence of taurine, indicating that taurine may be increasing ATP-dependent Ca2+ uptake through a mechanism of action involving the opening of cGMP-gated channels. The activation of cGMP-gated channels with dibutyryl-cGMP and with phosphodiesterase inhibition using zaprinast caused an increase in ATP-dependent Ca2+ uptake in isolated ROS, but not in taurine-stimulated ATP-dependent Ca2+ uptake. LY83583 had the same effects in isolated ROS as in retinal membrane preparations. Another inhibitor of cGMP-gated channels, Rp-8-Br-PET-cGMPS, produced the same pattern of inhibition in isolated ROS as LY83583. Thus, there appears to be a causal link between taurine and the activation of the cGMP-gated channels in the ROS under conditions of low calcium concentration, a connection that suggests an important role for taurine in the visual signalling function of the retina. PMID- 9871491 TI - Renal excretory responses of taurine-depleted rats to hypotonic and hypertonic saline infusion. AB - Male Wistar-Kyoto rats were given either tap water (control) or 3% beta-alanine (taurine-depleted) for three weeks. To prepare for the kidney function studies, the animals were then implanted with femoral vessels and bladder catheters. Two days after surgery, each rat was given an intravenous infusion of saline at the rate of 50 microliter/min and urine samples were collected at specific time intervals. An isotonic saline solution (0.9% NaCl) was infused for determination of baseline parameters and was followed by the infusion of a hypotonic saline solution (0.45% NaCl). Two days later, the infusion protocol was repeated in the same animals; however, a hypertonic saline solution (1.8% NaCl) was substituted for the hypotonic saline solution. Renal excretion of fluid and sodium increased in the control, but not taurine-depleted, rats during the hypotonic saline infusion. Interestingly, diuretic and natriuretic responses were similar between the groups during hypertonic saline infusion. The results suggest that taurine depletion in rats affects renal excretory responses to a hypotonic, but not a hypertonic, saline solution. PMID- 9871492 TI - Locally infused taurine, GABA and homotaurine alter differently the striatal extracellular concentrations of dopamine and its metabolites in rats. AB - We studied in vivo the effects of locally infused taurine (50, 150, and 450 mM) on the striatal dopamine and its metabolites in comparison with those of GABA and homotaurine, a GABAA receptor agonist, in freely moving rats. The extracellular dopamine concentration was elevated maximally 2.5-, 2- and 4-fold by taurine, GABA and homotaurine, respectively. At 150 mM concentration, at which the maximum effects occurred, homotaurine increased the extracellular dopamine more than taurine or GABA. When taurine and GABA were infused simultaneously with tetrodotoxin the output of dopamine did not differ from that in the presence of tetrodotoxin alone. In comparison, tetrodotoxin did not inhibit the increase in extracellular dopamine caused by homotaurine. Furthermore, omission of calcium from the perfusion fluid inhibited the increase of extracellular dopamine caused by GABA. However, it did not block the increase of dopamine caused by taurine or homotaurine. The present study suggests that the effects of intrastriatal taurine, GABA and homotaurine on the striatal extracellular dopamine differ. Thus, these amino acids seem to affect the striatal dopaminergic neurons via more than one mechanism. PMID- 9871493 TI - Shape and size changes induced by taurine depletion in neonatal cardiomyocytes. AB - Taurine is a very important organic osmolyte in most adult cells. Because of this property it has been proposed that large changes in the intracellular content of taurine can osmotically stress the cell, causing changes in its size and shape. This hypothesis was examined by measuring cell dimensions of taurine deficient cardiomyocytes using confocal microscopy. Incubation of isolated neonatal rat myocytes with medium containing 5 mM beta-alanine led to a 55% decrease in intracellular taurine content. Associated with the loss of taurine was a reduction in cell size. Two factors contributed to the change in cell size. First, there was a shift in cell shape, favoring the smaller of the two cellular configurations commonly found in the myocyte cell culture. Second, the size of the polyhedral configuration was reduced after beta-alanine treatment. These same two events also contributed to size reduction in cardiomyocytes incubated with medium containing 30 mM mannitol. Nonetheless, some qualitative differences exist between cells osmotically stressed by increasing the osmolality of the incubation medium and decreasing intracellular osmolality. The results support a role for taurine in the regulation of osmotic balance in the neonatal cardiomyocyte. PMID- 9871494 TI - Expression and localization of cysteine dioxygenase mRNA in the liver, lung, and kidney of the rat. AB - The expressions of cysteine dioxygenase (CDO) gene in the liver, lung, skeletal muscle, and kidney were studied by in situ hybridization with a cDNA probe from rat liver CDO under normal conditions. Significant expression of the CDO gene was detected in the liver, lung, and kidney, but not skeletal muscle. In the liver, the signal was confined to the cytoplasm of the hepatocytes. Furthermore, the signal was stronger in the periportal than that in the perivenous areas. In the lung, an intensive signal was found in the bronchiolar epithelium. As to the kidney, an intensive signal was observed in the distal convoluted tubules, while no signal was found in the proximal convultions. PMID- 9871495 TI - Immunohistochemical localization of taurine in various tissues of the mouse. AB - The localization of taurine was investigated in several tissues of the mouse. Immunohistochemical methods using a polyclonal antibody for taurine derived from rabbits was used in these studies. This method was used since it is a simple procedure and the results are clear and reliable. Tissues were fixed with paraformaldehyde, embedded in paraffin and treated in a microwave oven before using an avidin-biotin-complex method (ABC method). Control staining was accomplished by employing absorption staining using various amino acids: taurine, arginine, cysteine, hypotaurine and others. For purposes of comparison, radioautography (RAG) with 3H-taurine was performed to confirm the reliability of the immunohistochemical staining compared with the localization of the 3H-taurine incorporation in endothelial cells of the blood vessels of several tissues. In this investigation, immunoreactivity was broadly observed in many tissues: Purkinje cells of the cerebellum, glia cells of brain tissue, cardiac muscle cells, matrices of the bone, mucus granules of goblet cells of the intestines, and brown adipose cells of the fetus. Although the meaning of this widespread localization of taurine can not be explained completely, we surmise that taurine may have a different function in each of the tissues. In addition, taurine reactivity was observed in cell nuclei which was evidence of the presence of taurine in the nuclei. PMID- 9871496 TI - Is there a correlation between taurine levels and xenobiotic-induced perturbations in protein synthesis?: a study with tetracycline in rats. AB - Changes in urinary levels of taurine have been reported in rats following treatment with various xenobiotics including those which alter protein synthesis and/or are hepatotoxic. This paper reports on the time course of the urinary elevation of taurine following treatment of rats with tetracycline (50, 150 and 200 mg.kg-1). Maximum taurine excretion occurred 8-12 h following dosing. Serum albumin and total protein were significantly lower after 24 h (200 mg.kg-1). The increase in urinary taurine was dose-related and reflected in the raised serum levels of taurine 24 h after dosing. Serum and urinary protein and [3H]-leucine incorporation into acid precipitable protein in liver and muscle were reduced by tetracycline (100, 150 and 200 mg.kg-1) 10 h after dosing. The reduction in protein synthesis was correlated with increased urinary and serum levels of taurine at 10 h. The use of taurine as a non-invasive marker of protein synthesis is discussed. PMID- 9871497 TI - Effects of in vivo taurine depletion on induced-chemiluminescence production in macrophages isolated from rat lungs. AB - Alveolar macrophages isolated by pulmonary lavage from partially taurine-depleted rats demonstrated increased (2-2.6 fold) chemiluminescence due to the extracellular reaction between exogenous zymosan and various reactive forms of oxygen compared to macrophages isolated from control animals. Partial taurine depletion was achieved by adding 3% beta-alanine to the drinking water of the rats for 5 weeks prior to harvesting the macrophages. Superoxide dismutase activity was not increased in the lung tissue of the taurine-depleted rats. These data suggest that taurine has antioxidant properties and that taurine depletion is potentially deleterious to alveolar macrophages and pulmonary tissue. PMID- 9871498 TI - Changes of plasma insulin, urea, amino acids and rumen metabolites in somatotropin treated dairy cows. AB - An experiment was performed to evaluate the effects of somatotropin on plasma free amino acid, urea and insulin concentrations and rumen fermentation pattern and to assess their relationships. Four Italian Friesian dairy cows fitted with rumen cannulae were used in a switch-back design. Slow releasing recombinant bovine somatotropin (640 mg/cow) was injected every 28 days for two consecutive periods. Rumen fluid and blood samples were collected before and after feeding at 0, 7 and 21 days after rbST injection. Exogenous rbST increased plasma insulin concentration and the insulin response to feeding, and decreased plasma urea and free essential and branched chain amino acid concentrations. rbST did not affect rumen fermentation pattern. No correlation was found between rumen and plasma parameters measured after feeding. Our results are consistent with the notion that the main effect of somatotropin is post-absorptive. PMID- 9871499 TI - Taurine, glutamate and GABA modulate the outgrowth from goldfish retinal explants and its concentrations are affected by the crush of the optic nerve. AB - The amino acid taurine plays an important trophic role during development and regeneration of the central nervous system. Other amino acid systems, such as those for glutamate and gamma-aminobutyric acid (GABA), are modified during the same physiological and pathological processes. After crushing the optic nerve, goldfish retinal explants were plated in the absence and in the presence of different amino acids and amino acid receptor agonists. The length and the density of the neurites were measured at 5 days in culture. Taurine increased the length and the density of neurites. Glutamate and glycine increased them at low concentration, but were inhibitors at higher concentration. The combination of N methyl-D-aspartate (NMDA) and glycine produced a greater inhibitory effect than NMDA alone. NMDA or alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) added simultaneously with taurine impaired the stimulatory effect of the latter. GABA stimulated the emission of neurites in a concentration dependent manner. Hypotaurine also elevated the length of neurites, but cysteinsesulfinic acid did not produce a significant effect. The concentrations of taurine, glutamate and GABA were determined by HPLC with fluorescent detection in the retina of goldfish at various days post-crushing the optic nerve. The levels of taurine were significantly increased at 48h after the crush, and were elevated up to 20 days. Glutamate level decreased after the lesion of the optic nerve and was still low at 20 days. GABA concentration was not significantly different from the control. The interaction of these amino acids during the regenerative period, especially the balance between taurine and glutamate, may be a determinant in restoring vision after the crush. PMID- 9871500 TI - A comparative SAR study of thrombin receptor derived non peptide mimetics: importance of phenyl/guanidino proximity for activity. AB - Thrombin, the most potent physiological platelet agonist interacts with cells through a specific G protein-coupled receptor which has been cloned and sequenced. Synthetic thrombin receptor peptides (TRAPs) comprising the first 5 amino acids (SFLLR and SFLLR-NH2) of the new N-terminus tethered ligand of the thrombin receptor that is generated by thrombin's proteolytic activity were found to cause full platelet aggregation. During the screening of novel thrombin receptor derived non-peptide mimetics in the platelet aggregation assay we found that 1-phenylacetyl-4-(6-guanidohexanoyl)-piperazine (1) and 1-(6 guanidohexanoyl)-4-(phenylacetylamidomethyl)-piperidine (2) exerted in vitro antagonist activities (56% and 40% correspondingly) as it is depicted by the platelet aggregation assay. Using Molecular Modeling, the synthetic compounds were overlayed with SFFLR. All three superimposed low energy structures had Phe and Arg amino acids in spatial close proximity. The superimposition results revealed that 1 resembled more the stereoelectronic environment of SFLLR than 2. This difference may be related to their different antagonist efficacy. PMID- 9871501 TI - Maximal growth occurs at a broad range of essential amino acids to total nitrogen ratios in kittens. AB - Kittens fed diets containing 2.0 and 3.0 times (x) the NRC (1986) essential amino acid (EAA) requirement (EAArq) and 210 to 560 g crude protein (CP)/kg diet had growth rates and plasma amino acid patterns that were not significantly different than kittens fed a control diet (CD) containing 1.5 x EAArq and 350 g CP/kg diet. Growth rates of kittens fed diets containing only EAA (with nontoxic levels of arginine and methionine) and 280 to 460 g CP/kg diet were equivalent to those of kittens fed CD. Kittens fed only EAA and 140 and 210 g CP/kg diet had growth rates that were significantly lower than kittens fed CD. Since the growth rate of kittens fed 1.5 x EAArq and 210 g CP/kg diet in a previous experiment was equivalent to kittens fed CD (Taylor et al., 1997), it is suggested that the requirement for CP is higher (up to 280 g CP/kg diet) when only EAA are fed. The higher crude protein requirement appears to be primarily a consequence of the high obligatory nitrogen loss as urea (especially from arginine) incurred in the conversion of nitrogen from EAA to dispensable amino acids in the liver and secondarily because of a slow rate of catabolism of the EAA. A 3-dimensional plot of weight gains vs. CP levels and EAA to total nitrogen (E:T) ratios of kittens shows a broad range of CP levels and E:T ratios that support optimal growth in the kitten. It is suggested that similar patterns would occur in the chick, rat and other species if adverse effects caused by excesses of specific amino acids are avoided. PMID- 9871502 TI - Total plasma homocysteine as part of the routine aminogram by ion-exchange chromatography. AB - Ion-exchange chromatography (IEC) with ninhydrin post-column derivatisation is the only technique available for the assay of total, (free plus bound), cysteine and homocysteine which also enables the routine measurement of all other commonly occurring amino acids. IEC assay of total cysteine and homocysteine typically involves incubating buffered plasma for 60 minutes at 37 degrees C with dithiothreitol (DTT), but these assay conditions significantly extend total analysis time and compromise other amino acid values, notably glutamine and glutamate. However, it is possible to carry out the DTT reduction in plasma virtually instantaneously and without additional buffering, thus preserving the integrity of other diagnostically important amino acids. Assay precision is adequate for cardiovascular risk assessment. PMID- 9871503 TI - Effect of regular training on the myocardial and plasma concentrations of taurine and alpha-amino acids in thoroughbred horses. AB - Exercise induces significant changes in the free intracellular amino acid pool in skeletal muscle but little is known of whether such changes also occur in cardiac muscle. In this study the effect of regular exercise on the size and the constituents of the free amino acid pool in the hearts and in the plasma of thoroughbred horses was investigated. The total free intracellular amino acid pool in the hearts of control horses was 30.9 +/- 1.2 mumol/g wet weight (n = 6). Glutamine but not taurine was present at the highest concentration (13.5 +/- 0.9 and 7.7 +/- 0.69 mumol/g wet weight for glutamine and taurine respectively). As for the rest of the amino acids in the pool, only glutamate and alanine were present at levels greater than 1 mumol/g wet weight (4.6 +/- 0.25 and 1.7 +/- 0.14 for glutamate and alanine respectively). The tissue to plasma ratio was highest for taurine at 155, followed by glutamate at 111, aspartate and glutamine at 37, alanine at 5.8 and ratios of less than 3 for the rest of the amino acids. The total free intracellular amino acid pool in the hearts of exercised horses was slightly but not significantly lower than control (28.1 +/- 1.1 mumol/g wet weight, n = 6). Regular exercise increased the intracellular concentration of threonine, valine, isoleucine, leucine and phenylalanine but was only significant (p < 0.05) for threonine. This work has documented the profile of taurine and protein amino acids in the heart and in the plasma of thoroughbred horses and showed that in contrast to skeletal muscle, heart muscle does not show major changes in amino acids during regular exercise. PMID- 9871504 TI - Glycerol affects the quantification of aspartate and glutamate in acid-hydrolyzed proteins. AB - Glycerol is widely used in protein isolation pathways to improve folding and solubility of the proteins of interest. Amino acid composition analysis of protein samples hydrolyzed in the presence of glycerol resulted in underestimation of aspartate and glutamate, when compared to hydrolysis in the absence of glycerol. Quantification of free asparagine, aspartic acid, glutamine and glutamic acid hydrolyzed with hydrochloric acid or methanesulfonic acid in the presence of glycerol resulted in poor recoveries of aspartate and glutamate (between 6 and 66%). Gas chromatography-mass spectrometry analysis of the hydrolyzates revealed, as expected, the presence of esterification products. The esters were formed between the primary and secondary hydroxyl groups of the glycerol and both carboxyl groups of the amino acids. Protein samples intended for compositional analysis should be free of glycerol. PMID- 9871505 TI - Free D- and L-amino acids in ventricular cerebrospinal fluid from Alzheimer and normal subjects. AB - Free D-Ser, D-Asp and total D-amino acids were significantly higher (p < 0.05) in Alzheimer (AD) ventricular CSF than in normal CSF. There was no significant difference in the total L-amino acids between AD and normal CSF, but L-Gln and L His were significantly higher (p < 0.05) in AD-CSF. The higher concentrations of these D- and L-amino acids in AD ventricular CSF could reflect the degenerative process that occurs in Alzheimer's brain since ventricular CSF is the repository of amino acids from the brain. PMID- 9871506 TI - Chemo-enzymatic transformations in sensitive systems: lipase mediated hydrolysis of vancomycin esters. AB - Recently, an interest in the development of new vancomycin derivatives has been demonstrated. Here, the feasibility of using lipases, particularly those from Pseudomonas sp., for the hydrolysis of vancomycin alkyl esters is demonstrated. Benzyl ester derivatives were more easily cleavable than methyl ester derivatives, resulting in good yields of vancomycin acids without degradation. PMID- 9871507 TI - Cyclopentanedi- and tricarboxylic acids as squalene synthase inhibitors: syntheses and evaluation. AB - Based on earlier lead squalene synthase inhibitor A-87049 (3) and zaragozic acids, a series of cyclopentanedi- and tricarboxylic acids were synthesized and evaluated against the enzyme. Some exhibited good potency and SAR revealed the importance of conformation and substitution pattern of these synthetic inhibitors. PMID- 9871508 TI - Sulfonamide-based hydroxamic acids as potent inhibitors of mouse macrophage metalloelastase. AB - The structural requirements of sulfonamide-based hydroxamic acid 1 for inhibition of macrophage metalloelastase (MME) were investigated. A short aliphatic group at the R2 position together with an aromatic group at the R3 position significantly improved the inhibitory activity. Compounds 32, 34, and 40 were the most potent inhibitors of MME with IC50 values between 5 and 6 nM. PMID- 9871509 TI - Synthesis and in vitro antimalarial activity of sulfone endoperoxides. AB - A series of 4,8-dimethyl-4-phenylsulfonylmethyl-2,3-dioxabicyclo[3.3.1]+ ++nonanes, carrying a variety of substituents at position-8 (4) were prepared by a short and efficient method from R-(+)-limonene. Key reactions include thiol oxygen cooxidation, and alkylation and acylation of a sterically hindered tertiary alcohol compatible with the endoperoxy functionality. Some of compounds 4, which are structurally related to yingzhaosu A (2), were found to exhibit in vitro antimalarial activity comparable to that of artemisinin (1) and superior to that of arteflene (3). PMID- 9871510 TI - Synthesis and antitumor activity of leinamycin derivatives: modifications of C-8 hydroxy and C-9 keto groups. AB - A series of leinamycin derivatives were synthesized and evaluated for antitumor activity. Modifications at C-8 and C-9 positions revealed a broad structure activity relationship in vitro and some derivatives showed potent antiproliferative activity against HeLa S3 cells. PMID- 9871511 TI - Peptidyl human heart chymase inhibitors. 1. Synthesis and inhibitory activity of difluoromethylene ketone derivatives bearing P' binding subsites. AB - Peptidyl difluoromethylene ketone derivatives were designed to take advantage of probable additional interactions with the S' subsite of human heart chymase. They showed potent inhibitory activities against human heart chymase and were more efficient than bovine chymotrypsin. PMID- 9871512 TI - Peptidyl human heart chymase inhibitors. 2. Discovery of highly selective difluoromethylene ketone derivatives with Glu at P3 site. AB - Appropriate structural modification of the difluoromethylene ketone derivatives at both P3 and P' positions led us to the discovery of peptidyl human heart chymase inhibitor 12h which shows potent activity with Ki = 6 nM and high selectivity against closely related serine protease bovine alpha-chymotrypsin (chymotrypsin Ki = > 100 microM). Using the compound 12b, a docking study with human heart chymase was carried out to presume probable interactions. PMID- 9871513 TI - Synthesis and metabotropic glutamate receptor activity of a 2 aminobicyclo[3.2.0]heptane-2,5-dicarboxylic acid, a molecule possessing an extended glutamate conformation. AB - A photochemical approach to the synthesis of the aminobicycloheptane 6a is reported. This compound assumes an extended glutamate conformation, and for this reason was created to further probe the structural features relevant to achieving selectivity for the subtypes of the metabotropic glutamate family of receptors. PMID- 9871514 TI - Novel HIV-protease inhibitors containing beta-hydroxyether and -thioether dipeptide isostere surrogates: modification of the P3 ligand. AB - Studies involving modifications to the P3 position of previously described HIV protease inhibitors containing beta-hydroxyether and thioether dipeptide isostere replacements led to the discovery of pseudopeptides 8o and 8p with improved antiviral activities. PMID- 9871515 TI - Studies on an immunosuppressive macrolactam, ascomycin: synthesis of a C-33 hydroxyl derivative. AB - Ascomycin 2, a close analogue of the immunosuppressant FK506 1, was modified to incorporate a hydroxyl group at the C-33 position. This increased the aqueous solubility of ascomycin by a hundred-fold at pH 7.4 and by approximately 300-fold at pH 6.5. Ascomycin 3 also exhibited an excellent immunosuppressive activity in vitro, as tested in a human mixed lymphocyte proliferation (HuMLR) assay, and in vivo using a rat popliteal lymph node (rPLN) hyperplasia assay. PMID- 9871516 TI - Oligoribonucleotides containing 8-oxo-7,8-dihydroguanosine and 8-oxo-7,8-dihydro 2'-O-methylguanosine: synthesis and base pairing properties. AB - 7,8-Dihydroguanosine(8-oxoG) and 7,8-dihydro-2'-O-methylguanosine (8-oxoG-Me) were chemically synthesized. Oligoribonucleotides which contain 8-oxoG or 8-oxoG Me were synthesized. Incorporation of nucleotides opposite 8-oxoG and 8-oxoG-Me with dNTP by MMLV reverse transcriptase during cDNA synthesis was characterized. PMID- 9871517 TI - Peptide inhibitors of N-succinyl diaminopimelic acid aminotransferase (DAP-AT): a novel class of antimicrobial compounds. AB - Dipeptide substrates of N-Succinyl Diaminopimelic Acid Aminotransferase (DAP-AT) were converted to hydrazines by treatment with hydrazine and cyanoborohydride. These compounds were tested in vitro as inhibitors of DAP-AT from E. coli and in vivo as antibiotics. The hydrazino-dipeptides showed potent slow binding slow binding inhibition of DAP-AT as well as antimicrobial activity. PMID- 9871518 TI - Synthesis of beta-D-arabinofuranosyl-1-monophosphoryl polyprenols: examination of their function as mycobacterial arabinosyl transferase donors. AB - A convenient synthetic strategy has been developed to produce libraries of beta-D arabinofuranosyl-monophosphorylpolyprenol. Those containing C50 and C55 polyprenols were the most active as donors for the cell-free synthesis of the arabinans of mycobacterial cell walls. PMID- 9871519 TI - Phthalimidomethyl as a drug pro-moiety. Probing its reactivity. AB - Phthalimidomethyl derivatives 1, encompassing a wide range of leaving group abilities, are rapidly hydrolysed to the corresponding phthalamic acid via rate determining attack at the phthalimide carbonyl group. PMID- 9871520 TI - Peptidomimetic aminomethylene ketone inhibitors of interleukin-1 beta-converting enzyme (ICE). AB - Pyridone-based peptidomimetic inhibitors of recombinant human Interleukin-1 beta converting enzyme (ICE, caspase-1) with an aminomethylene ketone activating group in the P1' position are described. Several analogues with sub-nanomolar Ki's versus ICE and improved aqueous solubility are reported. PMID- 9871521 TI - Synthesis and SAR of a novel, potent and structurally simple LTD4 antagonist of the quinoline class. AB - The two geminal ethyl groups in the succinic acid moiety of CGP57698 (4-[3-(7 fluoro-2-quinolinyl-methoxy)phenyl-amino]-2,2-diethyl-4-oxo- butanoic acid) are responsible for the high in vitro and in vivo potency of this peptidoleukotriene antagonist of the quinoline type. The synthesis and structure activity relationships of CGP57698 and its analogs are described. PMID- 9871522 TI - 7,8-Methylenedioxy-4H-2,3-benzodiazepin-4-ones as novel AMPA receptor antagonists. AB - The synthesis and anticonvulsant activity of novel 7,8-methylenedioxy-4H-2,3 benzodiazepin-4-ones 3a-e, structurally-related to GYKI 52466 1, a well-known noncompetitive AMPA-receptor antagonist, are reported. The new compounds possess marked anticonvulsant properties and, in analogy to 1, antagonize seizures induced by AMPA. In addition, when compared to the model compound 1, compounds 3 show a longer-lasting anticonvulsant activity and a lower toxicity. PMID- 9871523 TI - A 2-methyleneoxetane analog of orlistat demonstrating inhibition of porcine pancreatic lipase. AB - The 2-methyleneoxetane analog 2 of orlistat (OLS, 1) has been synthesized and tested against porcine pancreatic lipase (PPL). Despite the loss of the carbonyl group, a potential site for hydrogen bonding interaction with the enzyme and the key element in the acylation by OLS, 2 has activity comparable to 1. PMID- 9871524 TI - Structure based design: novel spirocyclic ethers as nonpeptidal P2-ligands for HIV protease inhibitors. AB - A series of novel spirocyclic ethers were designed to function as nonpeptidal P2 ligands for HIV-1 protease inhibitors. Incorporation of designed ligands in the (R)-(hydroxyethylamino)sulfonamide isostere afforded potent HIV protease inhibitors. PMID- 9871526 TI - Syntheses and kinetic evaluation of hydroxamate-based peptide inhibitors of glyoxalase I. AB - Hydroxamate-containing tripeptide analogs resembling a reactive intermediate in glyoxalase I catalysis were prepared by solution methods and were found to be competitive inhibitors of the enzyme from Saccharomyces cerevisiae. Electronic properties of the hydroxamate functionality as well as those of the expected intermediates in the enzyme-catalyzed reaction were compared. PMID- 9871525 TI - Azepinoindole derivatives with high affinity for brain dopamine and serotonin receptors. AB - We synthesized 20 and 21 as conformationally constrained analogues of the dopamine receptor antagonist SKF-83742, as well as analogues 6-9, 16, and 18-22. Although 20 and 21 were inactive, 7, 9, and 19 showed strong binding to D-1, D-2, S-2, and alpha-1 receptors, as well as antipsychotic activity in vivo. PMID- 9871527 TI - Synthesis and evaluation of a photolyzable derivative of sphingosine 1-phosphate- caged SPP. AB - The synthesis of a photolyzable sphingosine 1-phosphate derivative is reported via the reaction of N-(tert-butoxycarbonyl)-2-N,3-O- isopropylidenesphingosine 7 and bis(alpha-methyl-o-nitrobenzyl) N,N-diisopropyl-phosphoramidite. Stimulation of DNA synthesis upon illumination of caged SPP-loaded cells is also described. PMID- 9871528 TI - Potent cyclic urea HIV protease inhibitors with 3-aminoindazole P2/P2' groups. AB - Cyclic ureas containing 3-aminoindazole P2/P2' groups are extremely potent inhibitors of HIV protease. The parent 3-aminoindazole 6 showed a Ki < 0.01 nM but poor translation of enzyme activity to antiviral activity was observed. A series of 3-alkylaminoindazoles revealed that translation improved with increasing lipophilicity. An X-ray crystal structure of 6 bound to HIV protease was obtained. PMID- 9871529 TI - Synthesis of the vancomycin CDE ring system. AB - A first generation synthesis of 22 is described constituting the first disclosure of the preparation of an appropriately protected and fully functionalized vancomycin CDE ring system complete with the C and E ring monochloro substitution pattern. The approach, which is based on two aromatic nucleophilic substitution reactions for sequential CD and DE macrocyclization, provided the opportunity to define and indirect solution to the control of the CDE atropisomer stereochemistry through selective thermal equilibration of the DE versus unaltered CD ring system. Its success provides a rationale for a preferred order to the CD and DE ring system introductions. PMID- 9871531 TI - A new antitumor agent: methyl sulfonium perchlorate of echinomycin. AB - Newly modified-echinomycin such as S-methylated sulfonium perchlorate of echinomycin (1), monosulfoxide (2), disulfoxid (3) and sulfone (4) have been prepared and evaluated for in vitro biological activities of cytotoxicity against P388, B16 and SNU-16 as well as in vivo antitumor activity against murine leukemia P388 and melanoma B16. PMID- 9871530 TI - Synthesis, SAR and pharmacology of CP-293,019: a potent, selective dopamine D4 receptor antagonist. AB - A series of novel, potent and selective pyrido[1,2-a]pyrazine dopamine D4 receptor antagonists are reported including CP-293,019 (D4 Ki = 3.4 nM, D2 Ki > 3,310 nM), which also inhibits apomorphine-induced hyperlocomotion in rats after oral dosing. PMID- 9871532 TI - Solid phase synthesis of benzylamine-derived sulfonamide library. AB - Using solid phase synthesis, a library has been constructed of benzylamine derived sulfonamides which have strong inhibitory activity against the blood coagulant thrombin. The library compounds were obtained in good yield and high purity; four of these thrombin inhibitors showed nanomolar potency (Ki 600-10 nM). PMID- 9871533 TI - Epoxide derivatives of pipecolic acid and proline are inhibitors of pipecolate oxidase. AB - The cis-4,5-epoxide derivative of L-pipecolic acid (2S,4S,5R-epoxypipecolic acid, cis-3) was synthesized and found to serve as an excellent substrate for L pipecolate oxidase (L-PO) and also to cause time-dependent, irreversible inactivation of the enzyme. Data are presented showing this compound is a mechanism-based inhibitor of L-PO, whereas 2S,3R,4S-epoxyproline acts as a reversible inhibitor. PMID- 9871534 TI - New nonsteroidal androgen receptor modulators based on 4-(trifluoromethyl)-2(1H) pyrrolidino[3,2-g] quinolinone. AB - A series of 2(1H)-pyrrolidino[3,2-g]quinolinones was prepared and tested for the ability to modulate the transcriptional activity of the human androgen receptor (hAR). The parent compound, 4-(trifluoromethyl)-2(1H)-pyrrolidino[3,2 g]quinolinone, displayed moderate interaction with hAR, but more substituted analogues, particularly 6,7-disubstituted compounds, were potent hAR agonists in vitro. PMID- 9871535 TI - On the preparative use of recombinant beta(1-3)galactosyl-transferase. AB - A number of non-natural N-acyl derivatives of glucosamine is incubated with a recombinant beta(1-3)galactosyl-transferase and UDP-galactose. Surprisingly, the enzyme recognizes the non-natural acceptors as substrates and transfers galactose onto the 3-OH group in a beta-mode to give a series of Lewis(c)-(type 1) disaccharides. PMID- 9871536 TI - Enzymatic synthesis of sialyl-Lewis(a)-libraries with two non-natural monosaccharide units. AB - A series of sialylated type-I sugars, which have the natural N-acetyl group of the glucosamine moiety replaced by a wide range of amides, is incubated with recombinant fucosyl-transferase III and non-natural guanosine-diphosphate activated donor-sugars. Surprisingly, the enzyme tolerates the simultaneous alterations on the donor and acceptor to form a wide array of sialyl-Lewis(a) analogues. PMID- 9871537 TI - Tripeptide growth hormone secretagogues. AB - A series of C-terminus capped dipeptides and tripeptides was synthesized as growth hormone (GH) secretagogues. Among them, tripeptide Aib-D-Trp-D-homoPhe-OEt showed low nanomolar activity in the rat pituitary assay. Thus, we have demonstrated that the GH secretagogue activity of the hexa-hepta-GH releasing peptides can be mimicked at the tripeptide level. PMID- 9871538 TI - (2S,4S)-amino-4-(2,2-diphenylethyl)pentanedioic acid selective group 2 metabotropic glutamate receptor antagonist. AB - (2S,4S)-2-Amino-4-(4,4-diphenylbut-1-yl)-pentane-1,5-dioic acid 1m, is a novel metabotropic glutamate receptor (mGluR) antagonist with insignificant ionotropic affinity. It is selective antagonist of negatively-coupled cAMP-linked mGluRs with no effect on phosphoinositide coupled mGluRs. A series of 4-substituted glutamic acid analogues were prepared and it was found that compound 1k is tenfold more potent than 1m. Compound 1k has neither significant affinity for ionotropic glutamate receptors nor group 1 and 3 metabotropic receptors. PMID- 9871539 TI - Identification of a non peptidic RANTES antagonist. AB - A series of phenothiazines demonstrating inhibition of RANTES binding to THP-1 cell membranes has been identified. The lead compound RP23618 (IC50 = 3 microM) was found to inhibit specific binding of 125I-RANTES, but not 125I-MCP-1 to THP-1 cell membranes and furthermore to antagonize RANTES, but not MCP-1-induced chemotaxis of THP-1 cells. PMID- 9871540 TI - Pyrazoles, 1,2,4-triazoles, and tetrazoles as surrogates for cis-amide bonds in boronate ester thrombin inhibitors. AB - Substituted pyrazoles, 1,2,4-triazoles, and tetrazoles are good surrogates for cis-amide bonds in a series of boronate ester thrombin inhibitors. PMID- 9871541 TI - Amygdalin binds to the CD4 receptor as suggested from molecular modeling studies. AB - The geometrical features of the proposed bioactive conformation of peptide T assessed by computational methods in a previous study, together with available structure-activity studies on peptide T, led us to propose a pharmacophore for the CD4-peptide T interaction. Subsequent, data base searching permitted us to identify amygdalin as a peptide T peptidomimetic. PMID- 9871543 TI - Synthesis of C2 alkynylated purines, a new family of potent inhibitors of cyclin dependent kinases. AB - The synthesis of a new family of inhibitors of the cell cycle regulating cyclin dependent kinases (CDK's) is reported. These compounds, related to the purines olomoucine and roscovitine, are characterised by the presence of alkynylated side chains at C2. They inhibit CDK's with IC50's in the 200 nM range. PMID- 9871542 TI - Affinity labelling of the tick-borne encephalitis virus RNA replicase proteins by 4-N-exo-base-substituted photoreactive CTP analogs. AB - 4-N-exo-base-substituted photoreactive analogs of CTP were designed and synthesized. Two flavivirus proteins NS5 and NS3 are shown to be labelled after RNA synthesis in the presence of the analogs, irradiation by UV-light (313 nm) and subsequent [alpha-32P]NTP incorporation. PMID- 9871544 TI - Synthesis and biological activity of 3-substituted 3-desoxynaltrindole derivatives. AB - The 3-unsubstituted and substituted analogs of naltrindole (NTI) were synthesized using palladium-catalyzed transformations, and their binding affinity to opioid receptors was determined. Although the 3-desoxy analog showed comparable delta selectivity with that of NTI, all of the novel compounds possessed low affinity for delta receptors indicating the important role of the 3-oxygen atom of NTI for interaction with delta-opioid receptors. PMID- 9871545 TI - Preparation of 9-deoxo-4"-deoxy-6,9-epoxyerythromycin lactams "motilactides": potent and orally active prokinetic agents. AB - A series of new, highly potent and orally active "motilactides", 9-deoxo-4"-deoxy 6,9-epoxyerythromycin lactams was designed, synthesized, and evaluated for their gastrointestinal motor stimulating activity. These compounds were acid stable and showed good oral efficacy. PMID- 9871546 TI - An approach to use an unusual adenosine transporter to selectively deliver polyamine analogues to trypanosomes. AB - In this paper we describe an approach to selectively deliver compounds to trypanosomes using an adenosine transporter which is unique to the trypanosome. Various polyamine analogues have been attached to known substrates of this adenosine transporter. The compounds prepared interact specifically with the adenosine transporter, some with a similar efficiency to berenil, a known substrate. PMID- 9871547 TI - L-374,087, an efficacious, orally bioavailable, pyridinone acetamide thrombin inhibitor. AB - Replacement of the amidinopiperidine P1 group of 3-benzylsulfonylamino-6-methyl-2 pyridinone acetamide thrombin inhibitor L-373,890 (2) with a mildly basic 5 linked 2-amino-6-methylpyridine results in an equipotent compound L-374,087 (5, Ki = 0.5 nM). Compound 5 is highly selective for thrombin over trypsin, is efficacious in the rat ferric chloride model of arterial thrombosis and is orally bioavailable in dogs and cynomolgus monkeys. The structural basis for the critical importance of both methyl groups in 5 was confirmed by X-ray crystallography. PMID- 9871548 TI - The synthesis and evaluation of cyclic ureas as HIV protease inhibitors: modifications of the P1/P1' residues. AB - Two series of cyclic ureas modified at the P1/P1' residue were prepared and evaluated for HIV protease inhibition and whole cell antiviral activity. Compounds 8b, 10 (3- and 4-pyridylmethyl analogs) and 6b (4-methoxy analog) showed significant improvement in antiviral activity relative to lead compounds DMP323 and DMP 450. PMID- 9871549 TI - Synthesis and pharmacological activity of the stereoisomers of GP-88, a propafenone-type modulator of multidrug resistance. AB - All four stereoisomers of the propafenone-type MDR-modulator GP-88 (1) were synthesized using a combined approach with chiral pool building blocks and an acetalic protective group, which allows not only diastereoseparation but also assignment of absolute configuration via NMR spectroscopy. Those isomers with different configuration on the center of chirality in the propanolamine side chain showed statistically different PGP-inhibitory activity. Generally, the (R) configured isomers were by a factor of nearby two higher active than the (S) isomers. No differences in activity were observed for isomers with different configuration on the benzylic center of chirality. PMID- 9871550 TI - Synthesis of a cosalane analog with an extended polyanionic pharmacophore conferring enhanced potency as an anti-HIV agent. AB - A novel cosalane analog having an extended polyanionic pharmacophore was synthesized in order to target specific cationic residues on the surface of CD4. The design rationale is based on a hypothetical binding model of cosalane to the surface of the protein. The new analog displayed an EC50 of 0.55 microM as an inhibitor of the cytopathic effect of HIV-1RF in CEM-SS cells, which represents a significant increase in potency over cosalane itself (EC50 5.1 microM). Both cosalane and the new analog are inhibitors of viral entry into target cells. PMID- 9871551 TI - Structure-based design and synthesis of a series of hydroxamic acids with a quaternary-hydroxy group in P1 as inhibitors of matrix metalloproteinases. AB - Examination of the S1 area of the active site of pro-stromelysin has led us to the design of novel and potent inhibitors of matrix metalloproteinases containing constrained quaternary-hydroxy group at P1. The synthesis and biological activity of these compounds with variations at P1', P2', and P3' will be described. PMID- 9871552 TI - Syntheses and evaluation of benzodiazaborine compounds against M. tuberculosis H37Rv in vitro. AB - The synthesis of six benzo[e]diazaborine compounds and thier in vitro evaluation against M. tuberculosis H37Rv is described. The compounds 1,2-dihydro-1-hydroxy-2 phenyl-2,4,1-benzo[e]diazaborin-3(4H)-one, 4, and 1,2-dihydro-1-hydroxy-2-(3 pyridyl)-2,4,1-benzo[e]diazaborin-3(4H) - thione, (5), showed the greatest inhibitory activity. PMID- 9871554 TI - Structural insight into the binding motifs for the calcium ion and the non catalytic zinc in matrix metalloproteases. AB - The binding motifs for the structural zinc and calcium ions in matrix metalloproteases (MMPs) were investigated by analyzing the three-dimensional structural models of 23 representative MMPs. PMID- 9871553 TI - Synthesis and antiviral activity of novel isodideoxy nucleosides with exocyclic methylene. AB - Novel isodideoxy nucleosides with exocyclic methylene were synthesized starting from L-xylose utilizing anomeric demethoxylation, Wittig reaction and Mitsunobu reaction as key steps and evaluated for antiviral activity. PMID- 9871555 TI - Irreversible inhibition of zinc-containing protease by oxazolidinone derivatives. Novel inactivation chemistry. AB - alpha-Benzyl-2-oxo-1,3-oxazolidine-4-acetic acid (BOOA) having (alpha R,4S) and (alpha S,4R) configurations were designed and synthesized as a novel type of mechanism-based inactivators for carboxypeptidase A (CPA), and kinetic analysis demonstrated that they indeed inhibit the enzyme in a time-dependent manner with the second order inhibitory rate constants (kinact/KI) of 1.52 and 1.39 M-1 s-1 for (alpha S,R4)-BOOA and (alpha R,4S)-BOOA, respectively. PMID- 9871556 TI - Hydroxyl radical-induced cross-linking of thymine and lysine: identification of the primary structure and mechanism. AB - Hydroxyl radical-induced formation of a cross-link of thymine (Thy) and lysine (Lys) in the gamma-radiolysis of N2O-saturated aqueous solution was studied. A Thy-Lys cross-link (I) of the formal structure that OH radical and 4-carbon centered Lys radical added respectively to C(5) and C(6) positions of Thy was isolated by a preparative HPLC and identified by a FAB-HRMS. The primary cross link I was dehydrated by treatment with HCl at 120 degrees C to yield the secondary structure (II) possessing a C(5)-C(6) double bond in the Thy moiety: the latter structure II was reported previously (Dizdaroglu, M.; Gajewski, E. Cancer Res. 1989, 49, 3463-3467). A pulse radiolysis study with a redox titration method indicated that 4-carbon centered Lys radical intermediate was of neutral redox reactivity in contrast to reducing reactivity of 5-hydroxy-5,6 dihydrothymin-6-yl radical intermediate. The cross-link I could be formed by a conventional radical recombination mechanism, but not by an ionic recombination mechanism involving a redox reaction between the radical intermediates. PMID- 9871557 TI - Organometallic photonucleases: a novel class of DNA-cleaving agents. AB - The first demonstration of DNA cleavage by an organic radical generated via homolysis of a metal-alkyl bond in a Cp-metal complex is presented. Irradiation of CpW(CO)3CH3 (1.5 molecules/base pair) produced methyl radical, giving single strand cleavage of pBR322 DNA. This process was inhibited by the general radical trap cysteine and by TEMPO, which traps carbon radicals but not oxygen-centered radicals. PMID- 9871558 TI - Effects of metal ions on the rates and enantioselectivities of reactions catalyzed by a series of semisynthetic transaminases created by site directed mutagenesis. AB - Fatty acid binding proteins are a class of small 15 kDa proteins with a simple architecture that forms a large solvent sequestered cavity. In previous work, we demonstrated that reductive amination reactions could be performed in this cavity by covalent attachment of a pyridoxamine cofactor and that the rate, enantioselectivity and substrate specificity of these reactions could be altered by site directed mutagenesis. Herein, we show that the chemistry performed by these conjugates can be extended to include catalytic transamination and describe the effects of added metal ions on reaction rate and enantioselectivity. We conclude that metal ions can be used to increase the rate of reactions catalyzed by semisynthetic transaminases; however, the addition of metal ions can also retard the reaction rate. Furthermore, it appears that the presence of metal ions almost always results in an erosion of reaction enantioselectivity. This limits their utility as a practical means of increasing reaction rate. The results reported here, for four independent systems, should be considered in future designs of artificial transaminases. PMID- 9871559 TI - A labeled guanidine ligand for studying sweet taste. AB - A synthesis of a biotinylated-, coumarin-substituted-N,N'-diarylguanidine 1 is reported. This ligand has structural features conducive to studying sweet taste including a fluorescent tag to facilitate spectroscopic studies of binding to protein-receptors for sweet ligands and an anchor for affinity purification. PMID- 9871560 TI - Synthesis and 5 alpha-reductase inhibitory activities of benzofuran derivatives with a carbamoyl group. AB - A series of 2-phenylbenzofuran derivatives with a diphenylmethylcarbamoyl group at the 5 or 6 position of the benzofuran ring were synthesized and evaluated for rat and human testosterone 5 alpha-reductase inhibitory activities in vitro. They had inhibitory activities against both enzymes and the 6-carbamoyl derivatives tended to be more potent than the 5-carbamoyl compounds. PMID- 9871561 TI - Synthesis and in vitro cancer cell growth inhibitory activity of monocyclic model compounds containing spongistatin triene side-chains. AB - Two monosaccharides embodying triene side-chains of the spongistatins display significant in vitro activity against human cancer cell lines. PMID- 9871563 TI - Synthesis and activity studies of N-[omega-N'-(adamant-1'-yl)aminoalkyl]- 2-(4' dimethylaminophenyl)acetamides: in the search of selective inhibitors for the different molecular forms of acetylcholinesterase. AB - A series of N-[omega-N'-(adamant-1'-yl)aminoalkyl]-2-(4'- dimethylaminophenyl)acetamides were synthesized and tested as acetylcholinesterase inhibitors. A significant selectivity toward acetylcholinesterases from various natural sources, mainly differing in their quaternary structure and solubility, was pointed out. The interest of this kind of molecules as potential therapeutic agents for Alzheimer's disease is discussed. PMID- 9871562 TI - 1-Methoxy-agroclavine from Penicillium sp. WC75209, a novel inhibitor of the Lck tyrosine kinase. AB - A high-throughput screen was developed and implemented to identify inhibitors of the Lck tyrosine kinase. This report describes the identification of a specific inhibitor of this enzyme from the solid fermentation culture of the Penicillium sp., WC75209. The active compound was isolated and structurally characterized as 1-methoxy-5R, 10S-agroclavine, a new member of the ergot alkaloid family. PMID- 9871564 TI - 1-[omega-(4-arylpiperazin-1-yl)alkyl]-3-diphenylmethylene-2,5- pyrrolidinediones as 5-HT1A receptor ligands: study of the steric requirements of the terminal amide fragment on 5-HT1A affinity/selectivity. AB - In the present paper, we report the synthesis and the binding profile on 5-HT1A, alpha 1 and D2 receptors of a new series of imide-arylpiperazines 3. The study of the length of the alkyl chain and the imide substructure allows us to suggest some important differences between the no-pharmacophoric sites of both 5-HT1A and alpha 1-adrenergic receptors, which could be of great importance in order to design new selective ligands. PMID- 9871565 TI - Synthesis of some hydroxynaphthazarins and their cardioprotective effects under ischemia-reperfusion in vivo. AB - A series of hydroxynaphthazarins has been synthesized. Some of them were found in in vivo experiments to be protectors of myocardium under ischemia-reperfusion and to reduce the infarction zone by 50% without any adverse effect. All compounds exhibit a moderate or small toxicity and are active in low doses. PMID- 9871566 TI - Enzymatic synthesis of a modified phospholipid and its evaluation as a substrate for B. cereus phospholipase C. AB - The novel phospholipid 2, which bears a tert-butyl moiety in place of the natural trimethyl ammonium group of phosphatidylcholine, has been enzymatically synthesized via a transphosphatidylation reaction mediated by phospholipase D. The change from the choline headgroup in 1 to the tert-butyl group in 2 reduced the efficiency of hydrolysis by the phosphatidylcholine-preferring phospholipase C from Bacillus cereus by a factor of greater than 10(3). PMID- 9871567 TI - A fast and inexpensive method for N-terminal fluorescein-labeling of peptides. AB - A new method has been developed to synthesize fluorescein labeled peptides, compounds of increasing importance in bioorganic chemistry, cell biology, pharmacology, drug targeting and medicinal chemistry. We show, that 4(5) carboxyfluorescein is much more efficient than the hitherto predominantly utilized reagents 4(5)-carboxyfluorescein-N-succinimidylester and 4(5) fluoresceinisothiocyanate. PMID- 9871568 TI - Selective photocleavage of single-stranded nucleic acids by cyclobisintercaland molecules. AB - Irradiation of mixtures of a single-stranded circular plasmid and of a double stranded supercoiled DNA in presence of the cyclobisintercaland compounds 2 or 3 shows that these reagents effect the selective photocleavage of the single stranded entity. Furthermore, 2 also cleaves tRNAasp preferentially at single stranded domains. PMID- 9871569 TI - Derivatization of soyasapogenol A and their hepatoprotective activities. AB - Fifteen derivatives of soyasapogenol A (1), which is another aglycon moiety of soyasaponins from soybean together with soyasapogenol B (2), were prepared and their in vitro hepatoprotective effects were evaluated. PMID- 9871570 TI - An endogenous sleep-inducing compound is a novel competitive inhibitor of fatty acid amide hydrolase. AB - 2-Octyl gamma-bromoacetoacetate (O gamma Br), an endogenous compound originally isolated from human cerebrospinal fluid (CSF), has previously been demonstrated to increase REM sleep duration in cats. Based on the chemical structure of O gamma Br and its reported sleep-inducing effects, we synthesized O gamma Br along with chemically related analogs and tested these compounds as inhibitors of fatty acid amide hydrolase (FAAH), a brain enzyme that degrades neuromodulatory fatty acid amides. O gamma Br was found to competitively inhibit FAAH activity with IC50 and Ki values of 2.6 microM and 0.8 microM, respectively [for the (R) enantiomer of O gamma Br (1)]. A set of synthetic analogs of O gamma Br was examined to define the structural features required for FAAH inhibition and inhibitor potencies were assessed at pH 9.0 (near the pH optimum of FAAH) and pH 7.0. Interestingly, at pH 7.0 the gamma-halo beta-keto ester inhibitors proved to be significantly more potent than the trifluoromethyl ketone of oleic acid, one of the most potent FAAH inhibitors described to date. This study supports the possibility that O gamma Br may be a physiological regulator of FAAH activity and fatty acid amide levels in vivo. Additionally, the characterization of gamma-halo beta-keto esters as powerful FAAH inhibitors near physiological pH may aid in future studies of the enzymology and biological properties of FAAH. PMID- 9871572 TI - Reactions of 1-methylbenzimidazole derivatives with m-chloroperoxybenzoic acid. AB - Oxidation of seven 1-methylbenzimidazole (MBI) derivatives (with pKa's ranging from 1.6 to 6.0) was carried out with m-chloroperoxybenzoic acid and structures of the products formed were identified. (Condensed benzene moiety-hydroxylated)-2 (m-chlorobenzyloxy)-MBIs and 2-oxo-MBIs were obtained from MBIs with pKa's of more than 5.6 and about 3.3, respectively. PMID- 9871571 TI - A novel series of N-(hexahydro-1,4-diazepin-6-yl) and N-(hexahydroazepin- 3 yl)benzamides with high affinity for 5-HT3 and dopamine D2 receptors. AB - A novel series of benzamides with a hexahydro-1,4-diazepine or hexahydroazepine ring in the amine moiety were prepared, and their binding affinities for 5-HT3 and dopamine D2 receptors were evaluated. The R isomer of the 1-ethyl-4 methylhexahydro-1,4-diazepinylbenzamide (R)-22 had potent affinity for both receptors. The R-enantiomer of the corresponding 1 ethylhexahydroazepinylbenzamide 28 showed potent affinity for dopamine D2 receptors with reduced affinity for 5-HT3 receptors, while the S isomer was found to be a potent and selective 5-HT3 receptor antagonist. PMID- 9871573 TI - Discovery of LB30057, a benzamidrazone-based selective oral thrombin inhibitor. AB - Systematic variation of the so-called P-pocket moiety of benzamidrazone-based selective thrombin inhibitors led to the discovery of LB30057. It is potent (Ki = 0.38 nM for human thrombin), selective (Ki = 3290 nM for bovine trypsin), and orally bioavailable (58% oral bioavailability in dogs). LB30057 was efficacious in thrombosis animal models. PMID- 9871574 TI - Solid phase organic synthesis of polyamine derivatives and initial biological evaluation of their antitumoral activity. AB - A series of N1-monosubstituted putrescine and spermine derivatives was synthesised using a solid phase methodology. We evaluated their cytotoxicity, calmodulin antagonism and polyamine uptake inhibition, pharmacological properties shared by some antitumoral agents. PMID- 9871575 TI - Molecular dynamics simulations of the structure of aldose reductase complexed with the inhibitor tolrestat. AB - This study reports a molecular dynamics (MD) investigation on the structure of aldose reductase (ALR2) complexed with the potent inhibitor tolrestat. The simulations predict four different orientations of tolrestat into the ALR2 binding site; these orientations have in common a strong interaction of the anionic carboxylate with Tyr48, His110, Trp111 and NADP+, but completely differ for the orientation of the aromatic portion of the inhibitor. Interestingly, the orientation in which tolrestat gives the most attractive interaction energy with the enzyme is in full accord with the x-ray crystal structure of the complex that has been reported in the literature after this work was completed. In addition, the suggestion of more than one orientation of tolrestat during MD is in agreement with recent electrospray mass spectrometry experiments on the ALR2 tolrestat complex. PMID- 9871576 TI - Toward the antibody-catalyzed chemiluminescence. Design and synthesis of hapten. AB - Hapten 4 was synthesized to generate catalytic antibodies triggering chemiluminescence by catalyzing the decomposition of the 1,2-dioxetane 3. The hapten 4 was so designed as to elicit a negatively charged functional group in the antibody combining site to catalyze the beta-elimination of the protecting group in 3 as well as to lock the protecting group into an energetically favorable anti-periplanar conformation. PMID- 9871577 TI - X-ray structural characterization of SR 142948, a novel potent synthetic neurotensin receptor antagonist. AB - SR 142948 is an original and extremely potent neurotensin receptor antagonist developed in a promising approach to novel antipsychotic drugs. The X-ray structure was elucidated and compared to SR 48692 and levocabastine, providing new informations about the possible recognition process of NT receptor subtypes. PMID- 9871578 TI - Syntheses of D- and L-myo-inositol 1,2,4,5-tetrakisphosphate and stereoselectivity of the I(1,4,5)P3 receptor binding. AB - D- and L-myo-Inositol 1,2,4,5-tetrakisphosphate [D- & L-I(1,2,4,5)P4], which are analogues of D-myo-Inositol 1,4,5-trisphosphate [D-I(1,4,5)P3], a calcium mobilizing second messenger, were synthesized via resolution of the camphanate ester of a myo-inositol derivative, and the binding affinities to I(1,4,5)P3 receptor were measured. PMID- 9871580 TI - Introduction of a nitroxide group on position 2 of 9-phenoxyacridine: easy access to spin labelled DNA-binding conjugates. AB - In the search for spin labelled intercalators of general use to construct DNA binding conjugates, 6-chloro-2-[(1-oxyl-2,2,5,5-tetramethyl- pyrrolin-3 yl)methyloxy]-9-phenoxy-acridine 5, has been prepared. This key-intermediate reacts with amines to give the corresponding labelled 9-amino substituted acridines. Comparative EPR and fluorescence measurements show that the label causes only little modification of the binding properties of acridine. PMID- 9871579 TI - Optimization of the binding properties of PNA-(5')-DNA chimerae. AB - The synthesis and evaluation of PNA-(5')-DNA chimerae containing either a 5' amide (i.e. 1a), a 5'-phosphodiester (i.e. 1b) or 5'-phosphonate linkages (i.e. 1c,d) at the junction site are described. The 5'-linkages could be installed using either 5'-amino-5'-deoxythymidine phosphoramidite 2, O-[2-(2-aminoethyl) (thymin-1-ylacetyl)amino]ethyl phosphoramidite 3, N-(2-aminoethyl)-N-(thymin-1 ylacetyl)aminomethyl phosphonate 4 or N-(2-aminoethyl)-N (allyloxycarbonyl)aminomethyl phosphonate 5 as building blocks, respectively. It is shown that PNA-(5')-DNA of type 1a-c have a higher binding affinity with complementary RNA than native DNA, and that the antisense activity is mainly due to RNase H. PMID- 9871581 TI - Dimerization of sumatriptan as an efficient way to design a potent, centrally and orally active 5-HT1B agonist. AB - A new bivalent ligand of formula 3 which results from the covalent coupling of two sumatriptan molecules with a p-xylyl spacer at the level of the sulfonamide nitrogen has been prepared and evaluated as a 5-HT1B/1D receptors agonist. In vitro experiments at 5-HT1B human cloned receptors (Ki = 0.64 nM; EC50 = 0.58 nM) and at the level of the contraction of the New Zealand white rabbit saphenous vein (pD2 = 6.6) demonstrate the superior potency of dimer 3 as a 5-HT1B receptor agonist when compared to sumatriptan or zolmitriptan. Interestingly enough, the new bivalent agonist 3 was found to induce hypothermia in the guineapig upon oral administration suggesting good oral activity and access to the brain. PMID- 9871582 TI - Synthesis and binding studies of a 1-alkyl-3,6-diamino-4-quinolone based receptor for N-acylated dipeptides. AB - The synthesis and binding properties of a semirigid host for N-acyldipeptide carboxylic acids is presented. The design is based on the rigidification of a peptide strand, coupled to the use of a substituted quinoline as a hydrogen bond acceptor for the proton of a carboxylic acid. PMID- 9871583 TI - Potent HIV protease inhibitors incorporating high-affinity P2-ligands and (R) (hydroxyethylamino)sulfonamide isostere. AB - Design and synthesis of a series of very potent nonpeptide HIV protease inhibitors are described. The inhibitors are derived from novel high affinity P2 ligands and (R)-(hydroxyethylamino)sulfonamide isostere. PMID- 9871585 TI - Diimidazo[1,2-c:4',5'-e]pyrimidines: N6-N1 conformationally restricted adenosines. AB - Tethering the N6-substituents of N6-substituted adenosines to N1 has resulted in a series of conformationally restricted adenosine analogues. The resultant diimidazo[1,2-c:4',5'-e]pyrimidines were shown to be adenosine A1 selective. PMID- 9871584 TI - Diimidazo[1,2-c:4',5'-e]pyrimidines: adenosine agonist activity demonstrated by microphysiometry. AB - Silicon-based microphysiometry, measuring extracellular acidification rate of cells in culture, demonstrated that a series of diimidazo[1,2-c:4',5' e]pyrimidines were agonists at the human adenosine A1 receptor. 5-amino-7,8 dihydro-3-ribofuranose-8-(R)-(phenyl)-3H-diimidazo [1,2-c:4',5'-e]pyrimidine (2a) had an EC50 of 100 microM and reached 90% of the Emax produced by R-PIA. PMID- 9871586 TI - Endothiopeptide inhibitors of HIV-1 protease. AB - Endothiopeptide inhibitors of HIV-1 protease were synthesized by chemical and enzymatic methods to individually replace each backbone amide bond in 1 with a thioamide-linkage. Interestingly, agent 7, which contains a thioamide-linkage between the P2' and P3' positions of 1, was the most potent, competitive inhibitor of HIV-1 protease with a Ki of 3.4 microM. PMID- 9871587 TI - Solid phase synthesis of a biased mini tetrapeptoid-library for the discovery of monodentate ITAM mimics as ZAP-70 inhibitors. AB - The biased library was composed of a novel phosphotyrosine mimic fixed in the P1 position of a tetrapeptoid and combined with three lipophilic N-substituents at the remaining positions giving a total of 27 single compounds. Screening for ZAP 70 antagonism identified 8 as a novel selective monodentate ZAP-70 antagonist and lead in the search for new immunosuppressive drugs. PMID- 9871588 TI - Structure activity relationship study of known inhibitors of the enzyme 5 alpha reductase (5AR). AB - Preliminary results of a modelling study of both steroidal and non-steroidal inhibitors of 5 alpha-reductase (5AR) are described in order to elucidate the essential structural requirements needed for the design of novel non-steroidal inhibitors. The study suggests that: (i) there is a requirement for groups to mimic the C(3) = O of the steroid substrate A-ring; (ii) the area of the active site about the C(17)-OH position of the substrate does not appear to possess hydrogen bonding groups and is unrestricted. PMID- 9871589 TI - Synthesis of a new class of camptothecin derivatives, the long-chain fatty acid esters of 10-hydroxycamptothecin, as a potent prodrug candidate, and their in vitro metabolic conversion by carboxylesterases. AB - Five (20S)-10-hydroxycamptothecin derivatives carrying the long-chain fatty acid esters were prepared for the development of a new class of prodrug-type agents. In vitro experiments using three kinds of purified carboxylesterase isozymes from the liver microsomes of rat, pig, and human demonstrated that these derivatives were efficiently metabolized by enzymes compared with CPT-11. PMID- 9871590 TI - Interpreting the effect of methyl group at the three carbon bridge of (-) huperzine A on its anticholinesterase activity by molecular dynamics method. AB - Based on the recently resolved crystal structure of complex (-)-huperzine A-AChE, we simulated the interaction between (-)-huperzine A analogues and AChE using molecular dynamics method. It was revealed that the methyl group at the three carbon bridge of (-)-huperzine A can form a weak hydrogen bond with the phenol hydroxyl oxygen of Tyr121 and the main-chain oxygen of Gly118 of AChE, respectively. PMID- 9871591 TI - Stereoselective synthesis of 11Z-9-demethyl-9-benzyl- and 9-phenyl-retinals and their interaction with bovine opsin. AB - 11Z-9-Demethyl-9-benzyl- and 9-phenyl-retinals were synthesized stereoselectively from the beta-ionone analog-tricarbonyliron complexes and their interaction with bovine opsin was investigated. PMID- 9871592 TI - Synthesis and cytotoxic activity of novel 10-alkylated docetaxel analogs. AB - An alkylation method of docetaxel at the C-10 position has been established by a radical coupling reaction using a 10-xanthate derivative of 7-O-TES-10 deacetylbaccatin III and appropriate alkenes. In addition the cytotoxic activity of 10-alkylated docetaxel analogs was evaluated. Among these analogs, a derivative having a methoxycarbonyl group at the end of the alkyl moiety exhibited more potent cytotoxic activity than docetaxel. PMID- 9871593 TI - Solid phase synthesis of N-carboxy alkyl-containing peptides derived from enantiopure alpha-keto-beta-aminoacids. AB - alpha-Keto-beta-aminoacids 5a-c can be reductively aminated with the peptide sequence H2N-Leu-Val-Phe-Phe on a solid support to afford N-carboxy alkyl peptides 1a-c. The N-carboxy alkyl lysine derivative 7 was subsequently extended from the N-terminus with glutamine and histidine residues. PMID- 9871594 TI - Synthetic arabinofuranosyl oligosaccharides as mycobacterial arabinosyltransferase substrates. AB - A series of arabinofuranosyl oligosaccharides found as constituent parts of the polysaccharide portion of the cell wall of Mycobacterium tuberculosis have been chemically synthesized. Screening of these oligosaccharides as substrates for arabinosyltransferases present in mycobacterial membrane preparations suggests that modified oligosaccharide analogs as small as disaccharides may be inhibitors of glycan biosynthesis. Such inhibitors would be of potential utility as lead compounds in the identification of new drugs for the treatment of mycobacterial infections. PMID- 9871595 TI - Bis(5-hydroxy-2-hydroxymethyl-pyran-4-one-6-yl)methane: a novel ligand for the intracellular mobilisation of ferritin-bound iron. AB - The efficacy of a novel tetradentate iron(III) ligand for the in vitro mobilisation of ferritin-bound iron is measured in direct comparison to the clinically approved agents, 1,2-dimethyl-3-hydroxypyridin-4-one and desferrioxamine. PMID- 9871596 TI - alpha-substituted quisqualic acid analogs: new metabotropic glutamate receptor group II selective antagonists. AB - Syntheses of both the alpha-methyl and benzyl analogs of quisqualic acid are described. Testing of these compounds for their activity at excitatory amino acid receptors revealed a striking change in activity in comparison to quisqualic acid. This structural modification results in the loss of quisqualate's potent agonist action at both non-NMDA ionotropic glutamate receptors as well as at group I mGluRs, while allowing these analogs to acquire antagonist properties with relative selectivity for group II metabotropic glutamate receptors. PMID- 9871597 TI - A series of non-quinoline cysLT1 receptor antagonists: SAR study on pyridyl analogs of Singulair. AB - The structure-activity relationship of a series of styrylpyridine analogs of MK 0476 (montelukast, Singulair) is described. This work has led to the identification of a number of potent and orally active cysLT1 receptor (LTD4 receptor) antagonists including 2ab (L-733,321) as an optimized candidate. PMID- 9871598 TI - Anti-malarial activities of acylated bruceolide derivatives. AB - Several O-acylated derivatives of bruceolide (2) were synthesized and their anti malarial activities together with selective toxicities were examined. It was found that 3,15-di-O-acetyl-(3c), 3,15-di-O-propionyl-(3d) and 15-O propionylbruceolide (3b), as well as bruceine B (3a), exhibited potent anti malarial activities with high selective toxicities. PMID- 9871599 TI - Synthesis of 2,3,4a,11b-tetrahydro-oxazino[2,3-c]benzopyran-9-carbonitriles as ATP-sensitive potassium channel openers. AB - A series of optically active tetrahydro-oxazino[2,3-c]benzopyran derivatives have been synthesized and evaluated for potassium channel opening activity. (4aR,11bR) 1-Benzoyl- 5,5-dimethyl-2,3,4a,11b-tetrahydro-oxazino[2,3-c]benzopyran-9-carb onitrile ((-)-11e) was identified as a bladder-selective potassium channel opener (IC50, bladder = 8.15 microM, IC50, portal vein = 34.5 microM). PMID- 9871600 TI - Functionalized 1,3-teraryls as a new class of hepatoprotectants. Part V. AB - Functionalized 1,3-teraryls, synthesized through ring transformation of 6-aryl-3 carbomethoxy-4-methylthio-2H-pyran-2-one from arylketone have been screened for their hepatoprotective activity and of them have demonstrated significant protection in animal model. PMID- 9871601 TI - Structure-activity relationships of a series of 1-substituted-4 methylbenzimidazole neuropeptide Y-1 receptor antagonists. AB - The characterization of a novel series of NPY-1 receptor antagonists derived from the 4-methylbenzimidazole 4 is described. Appropriate substitution on the piperidyl nitrogen of 4 led to systematic increases in Y-1 receptor affinity, to approximately 50-fold, and to the discovery of the importance of a second basic substituent. PMID- 9871603 TI - Nonpeptide glycoprotein IIb/IIIa inhibitors: substituted quinazolinediones and quinazolinones as potent fibrinogen receptor antagonists. AB - The synthesis and biological activity of a series of 3,6-substituted quinazolinediones and quinazolinones are described. The potent activity of these compounds as platelet aggregation inhibitors demonstrates the utility of these structures as central templates for nonpeptide RGD mimics. PMID- 9871602 TI - New bronchodilators selected by molecular topology. AB - Molecular topology has been applied to find new lead compounds with bronchodilator activity. Among the selected compounds stands out 3-(1H-tetrazol 5yl)-9H-thioxanthene-9 -one-10,10-dioxide, anthrarobin, 9-oxo-9H-thioxantene-3 carboxylic-10,10-dioxide acid, acenocoumarol and griseofulvin, with a percentage of relaxation, at 0.1 mM, of 91, 92, 85, 69, and 74%, respectively. Theophylline shows a correspondent value of 77% (Emax = 100% at 1 mM). PMID- 9871604 TI - Synthesis of a potent wide-spectrum serotonin-, norepinephrine-, dopamine reuptake inhibitor (SNDRI) and a species-selective dopamine-reuptake inhibitor based on the gamma-amino alcohol functional group. AB - A series of gamma-amino alcohols were synthesized and screened for reuptake inhibition and noncompetitive NMDA antagonism. Compound (+/-)-3f simultaneously and potently inhibits reuptake of 5-HT, NE, and DA, representing a potential wide spectrum reuptake inhibitor antidepressant. In addition, comparative rat and human studies uncovered a species-selective DA reuptake inhibitor (+/-)-2e, KD(hDAT)/KD(rDAT) = 97. PMID- 9871605 TI - 5-Aminomethylquinoxaline-2,3-diones, Part III: Arylamide derivatives as highly potent and selective glycine-site NMDA receptor antagonists. AB - A series of quinoxaline-2,3-diones with very high affinity to the glycine site of the NMDA receptor has been discovered. In contrast to the 7-nitro derivatives, the most potent 7-bromo substituted compounds were highly selective for the glycine site. Although none of the described compounds were active in the electroshock model in mice, 1a displayed significant protection in the quinolinic acid-induced excitotoxicity model in vivo. PMID- 9871606 TI - Recognition of 2'-deoxyisoguanosine triphosphate by HIV-1 reverse transcriptase and mammalian cellular DNA polymerases. AB - HIV-1 reverse transcriptase (RT) incorporates 2'-deoxyisoguanosine triphosphate (d-isoGTP) opposite thymidine (T) in a DNA template and opposite uracil (U) in an RNA template about 10 times more efficiently than the eukaryotic DNA polymerase alpha, both in the absence and presence of dATP. PMID- 9871607 TI - Phosphodiesterase inhibitory properties of losartan. Design and synthesis of new lead compounds. AB - A 4-centre PDE4 pharmacophore search has been carried out in several 3D-databases containing compounds belonging to different therapeutic areas. Losartan, an angiotensin-II antagonist, has been identified as a new lead compound for developing PDE4 inhibitors. New families of compounds derived from losartan has been synthesized and their PDE inhibition has been measured. PMID- 9871608 TI - Solid-phase synthesis of peptidomimetic oligomers with a phosphodiester backbone. AB - An unnatural biopolymer is described in which amino acid side-chains are presented along a negatively charged phosphodiester backbone. For this purpose, a series of phosphoramidite monomers was prepared from chiral 1,2-diols. These were efficiently converted into oligomers using standard coupling conditions on an automated DNA synthesizer. PMID- 9871609 TI - Novel bisphosphonate inhibitors of phosphoglycerate kinase. AB - A series of novel, conformationally-restrained bisphosphonate analogues of 1,3 bisphosphoglyceric acid 1 have been synthesised and evaluated as inhibitors of 3 PGK. They are competitive inhibitors of the human enzyme and, especially for certain alpha-halophosphonic acid analogues, both Ki and IC50 values extend into the submicromolar range. PMID- 9871610 TI - 2'-Deoxy-2'(S)-ethinyl oligonucleotides: a modification which selectively stabilizes oligoadenylate pairing to DNA complements. AB - Oligonucleotides consisting of 2'-deoxy-2'(S)-ethinyl-thymidine,-uridine, and adenosine have been prepared. Whereas the modified pyrimidine oligonucleotides uniformly lead to weaker pairing affinity with DNA and RNA complements, the corresponding adenine oligonucleotides show enhanced thermal stability in duplexes with complementary DNA and decreased stability with RNA. PMID- 9871611 TI - Novel quinolone derivatives as potent antibacterials. AB - Several 7-(3R,4R-N,N'-dialkyl diaminopyrrolidinyl)-substituted quinolones were synthesized and evaluated for antibacterial activities. 5-Amino-7-(3R,4R-N,N' dimethyldiamino-6,8-difluoro-1,4-dihydro-1-c yclopropyl -4-oxoquinoline-3 carboxylic acid was found to have potent antibacterial activity against gram +ve organisms. PMID- 9871612 TI - Discovery of a new cyclooxygenase-2 lead compound through 3-D database searching and combinatorial chemistry. AB - Using a combination of computational and combinatorial chemistry methodologies, a phenothiazine compound was discovered that is a selective inhibitor of cyclooxygenase-2 and serves as a lead compound for a potentially novel series of anti-inflammatory compounds. PMID- 9871613 TI - DNA cleavage by 7-methylbenzopentathiepin: a simple analog of the antitumor antibiotic varacin. AB - The compound 7-methylbenzopentathiepin, a simple analog of the benzopentathiepin antitumor antibiotic varacin, was shown to be a potent thiol-dependent DNA cleaving agent. Biological experiments previously suggested that DNA cleavage might play a role in the cytotoxicity of varacin; however, this is the first direct evidence that benzopentathiepins can cause DNA strand breaks under physiologically relevant conditions. PMID- 9871614 TI - Use of the 1,2,5-thiadiazolidin-3-one 1,1 dioxide and isothiazolidin-3-one 1,1 dioxide scaffolds in the design of potent inhibitors of serine proteinases. AB - The attachment of a phosphate leaving group to the 1,2,5-thiadiazolidin-3-one 1,1 dioxide and isothiazolidin-3-one 1,1 dioxide scaffolds was found to yield highly potent, time-dependent inhibitors of human leukocyte elastase (HLE). PMID- 9871615 TI - Indolequinone antitumor agents: relationship between quinone structure and rate of metabolism by recombinant human NQO1. AB - A series of indolequinones bearing various functional groups has been synthesized, and the effects of substituents on the metabolism of the quinones by recombinant human NAD(P)H:quinone oxidoreductase (NQO1), and on the toxicity toward nonsmall cell lung cancer cells with either high NQO1 activity (H460) or with no detectable activity (H596) were studied. PMID- 9871616 TI - Synthesis and antibacterial activity of O-methyl derivatives of azalide antibiotics: I. 4", 11 and 12-OMe derivatives via direct methylation. AB - A series of O-Me derivatives of 9-deoxo-8a-aza-8a-homoerythromycin has been prepared and evaluated for antibacterial activity. The relative rates of methylation of the four available hydroxyls (4", 6, 11 and 12) in 2',3'-bis-Cbz protected 9-deoxo-8a-aza-8a-homoerythromycin were compared to those given in a published report for the similarly protected 9a-azalide. An incongruity in the results prompted reinvestigation of the O-methylation of the 9a-azalide, and an error in structure assignment in the published report was discovered: the compound reported as the 6-OMe-9a-azalide has been determined to be the 12-OMe derivative. PMID- 9871617 TI - Modifications of the 4,4'-residues and SAR studies of Biphalin, a highly potent opioid receptor active peptide. AB - Modifications of 4,4' residues of Biphalin have resulted in greater binding selectivity and biological potency for the mu opioid receptor. A higher partition coefficient across the phospholipid bilayer membrane has been achieved by using a beta-branched unusual amino acids. PMID- 9871618 TI - Chemical inducers of dimerization: the atomic structure of FKBP12-FK1012A-FKBP12. AB - A ligand that simultaneously binds two proteins must have two high affinity protein binding domains joined in a fashion that facilitates, or at least does not prevent, protein-protein interaction. Designing such ligands is challenging, and a high resolution X-ray structure of FKBP12-FK1012A-FKBP12 illustrates the subtleties of one successful design. PMID- 9871619 TI - Assessment of the steric tolerance of the P sector in the catalytic site of porcine liver esterase. AB - Porcine liver esterase catalyzes the hydrolysis of simple esters capable of hydrogen bonding more rapidly than the reaction for small hydrophobic acetates. The result can be understood in terms of the high steric tolerance of the P sector in the catalytic site. PMID- 9871620 TI - Endothelin antagonists: search for surrogates of methylendioxyphenyl by means of a Kohonen neural network. AB - The methylendioxyphenyl group, present in a number of potent endothelin receptor antagonists, could have undesirable metabolic interactions with cytochrome P450 in vivo. Using a self-organizing neural network we analysed the features of molecular electrostatic potentials of several endothelin receptor ligands. A library of small "fragments and functional groups" together with their corresponding Kohonen maps was generated. By means of this Kohonen map library we discovered the benzothiadiazole group as a surrogate for methylendioxyphenyl. PMID- 9871621 TI - Endothelin antagonists: evaluation of 2,1,3-benzothiadiazole as a methylendioxyphenyl bioisoster. AB - The methylendioxyphenyl group is present in a number of endothelin receptor antagonists thus far reported. By means of a Kohonen neural network we discovered with a benzothiadiazole a bioisosteric replacement instead. This group should be devoid of the negative metabolic interactions with cytochrome P450 ascribed to methylendioxyphenyl in vivo. The synthesis of a potent benzothiadiazole analogue EMD 122801 together with in vitro studies of different methylendioxyphenyl, benzothiadiazole and benzofurazan derivatives is described. PMID- 9871622 TI - Hydroxamate-based inhibitors of low affinity IgE receptor (CD23) processing. AB - A series of hydroxamic acids related to the non-selective matrix metalloprotease inhibitor Batimastat is described, which inhibits the proteolytic cleavage of the low affinity IgE receptor from cell membrane preparations. Limited SAR studies suggest that the structural requirements for effective inhibition are distinct from those required for the inhibition of collagenase. PMID- 9871624 TI - Sugar-substituted 2-azetidinones as cholesterol absorption inhibitors. AB - The asymmetric synthesis of a glucuronide conjugate of the 2-azetidinone cholesterol absorption inhibitor Sch 48461 was accomplished to confirm the structure of a metabolite isolated from in vivo sources. Key features of this article include the asymmetric synthesis of 2-azetidinones by Evan's chiral oxazolidinone methodology and glucuronide formation by a Mitsunobu protocol. PMID- 9871623 TI - Selective inhibition of low affinity IgE receptor (CD23) processing. AB - A series of hydroxamic acids related to the non-selective matrix metalloproteinase inhibitor Batimastat has been prepared, some members of which are potent inhibitors of the processing of the low affinity IgE receptor (CD 23). Increased activity is obtained by appropriate substitution at the alpha-position, whilst selectivity is gained by use of a P1' benzyl group in conjunction with a C terminal primary amide. PMID- 9871625 TI - Synthesis and biological evaluation of 3-arylisoquinolines as antitumor agents. AB - To investigate the structure-activity relationship of 7,8-dimethoxy-2- methyl-3 (4,5-methylenedioxy-2-vinylphenyl)isoquinolin-1(2H) -one 2, diverse substituted 3 arylisoquinolines were synthesized and tested in vitro antitumor activity against five human tumor cell lines. The results showed a broad antitumor spectrum for a series of 3-arylisoquinolines. PMID- 9871626 TI - Synthesis of bisindolylmaleimides using a palladium catalyzed cross-coupling reaction. AB - Bisindolylmaleimides are known to be potent and selective PKC inhibitors. A new synthesis of this class of compound is reported. The key step is a Suzuki cross coupling reaction using a readily available indolylmaleimide triflate intermediate. PMID- 9871627 TI - High affinity phenylglycinol-based NK1 receptor antagonists. AB - Heterocyclic replacements for the carboxamido group of the previously disclosed phenylglycinol-based human NK1 (hNK1) receptor antagonists have been investigated, ultimately leading to acyclic compounds with sub-nanomolar affinity for the hNK1 receptor. PMID- 9871628 TI - Bis-S-acyl-2-thioethyl (SATE)-bearing monophosphate prodrug of beta-L-FD4C as potent anti-HBV agent. AB - The S-acyl-2-thioethyl (SATE)-bearing 5'-monophosphate prodrug of beta-L-FD4C (8) was synthesized and evaluated for its activity against HBV in the 2.2.15 cell line. This pronucleotide (8) exhibited an excellent inhibitory effect against HBV with an EC50 value that is more than eight fold lower than that of the parent nucleoside (4) under some assay conditions. It is also important to note that pronucleotide (8) was capable of inhibiting HBV replication by 90%; whereas its parent, beta-L-FD4C (4), could only inhibit virus replication no greater than 70% in the same assay. When evaluated in the standard cytotoxicity assay in CEM cell line, pronucleotide (8) exhibited an IC50 value of 52 microM, which was four times less toxic than parent beta-L-FD4C (4) (IC50 = 13 microM). PMID- 9871629 TI - Preparation of alpha-keto ester enol acetates as potential prodrugs of human neutrophil elastase inhibitors. AB - Enol acetates of a-keto esters with E configuration were prepared as potential prodrugs for human neutrophil elastase (HNE) inhibitors. PMID- 9871630 TI - 5-Aminomethylquinoxaline-2,3-diones. Part I: A novel class of AMPA receptor antagonists. AB - A series of 5-aminomethylquinoxaline-2,3-diones have been identified as potent and selective AMPA antagonists. Some of these compounds are also active at the glycine-binding site of the NMDA receptors. A number of these novel, water soluble quinoxaline-2,3-dione derivatives display protective effects in the electroshock-induced convulsion model in mice. PMID- 9871631 TI - 5-Aminomethylquinoxaline-2,3-diones. Part II: N-aryl derivatives as novel NMDA/glycine and AMPA antagonists. AB - Potent antagonists at the glycine-binding site of NMDA receptors, as well as dual antagonists acting also at AMPA receptors have been identified in a series of 5 arylaminomethylquinoxaline-2,3-diones. A study of the structure-activity relationship of these compounds is reported here. PMID- 9871632 TI - Discovery of FR115092: a novel antinephritic agent. AB - A series of dapsone-related 4-aminopheynl and 2-aminothiazolyl derivatives was prepared, and their antinephritic activity and blood toxicity were evaluated. 5 (2-Pyridylsulfonyl)-2-thiazolamine (FR115092, 26) was effective against two nephritis models, namely graft-versus-host disease (GVHD) and autoimmune W/BF1 mice, and showed none of the blood toxicity observed with dapsone. PMID- 9871633 TI - Synthesis, X-ray crystal structure, and biological activity of FR186054, a novel, potent, orally active inhibitor of acyl-CoA:cholesterol O-acyltransferase (ACAT) bearing a pyrazole ring. AB - The synthesis, single crystal X-ray structural analysis, and biological activity of FR186054 (2c), a new, potent, orally efficacious inhibitor of acyl CoA:cholesterol O-acyltransferase (ACAT), containing a pyrazole ring are described. This compound displayed excellent in vitro efficacy, irrespective of dosing method, indicating superior characteristics compared to other ACAT inhibitors. PMID- 9871634 TI - Antibiotic inhibitors of the peptidyl transferase center. 1. Clindamycin as a composite analogue of the transfer RNA fragments L-Pro-Met and the D-ribosyl ring of adenosine. AB - Clindamycin's three-dimensional structure is shown via a computer rendered stereomodel to be strikingly similar to L-Pro-Met and the D-ribosyl ring of adenosine. This discovery has important implications for the rational design of new licosamides and for efforts to understand how this and related classes of agents selectively inhibit protein biosynthesis on the prokaryotic ribosome. PMID- 9871635 TI - Novel antiproliferative falcarindiol furanocoumarin ethers from the root of Angelica japonica. AB - Four novel antiproliferative furanocoumarin ethers of falcarindiol, named japoangelols A (8.5), B (7.2), C (7.4), and D (8.4), were isolated from the root of Angelica japonica together with panaxynol (0.3), falcarindiol (3.2), (9Z)-1,9 heptadecadiene-4,6-diyne-3,8,11-triol (2.2), and 8-acetoxyfalcarinol (3.2). Structures were established from the spectroscopic evidence, and the inhibitory activities (ED50, microgram/ml, shown in the parentheses) were evaluated using the MTT assay. PMID- 9871637 TI - Investigations of azapeptides as mimetics of Leu-enkephalin. AB - Solution syntheses of azapeptide pentamers 2, 3, and 4 were accomplished. The binding affinity of these azapeptides and azatide 1 were examined in the context of monoclonal antibody 3-E7 known to strongly bind the [Leu5]enkephalin sequence. PMID- 9871636 TI - DNA gyrase inhibitory activity of ellagic acid derivatives. AB - Ellagic acid was found to inhibit E. coli DNA gyrase supercoiling with approximately the same potency as nalidixic acid. Tricyclic analogs of ellagic acid, which vary in the number and position of the hydroxy groups as well as their replacement with halogens, have been synthesized. The biological activity of these analogs is discussed. PMID- 9871638 TI - Synthesis, DNA-damaging and cytotoxic properties of novel topoisomerase II directed bisantrene analogues. AB - New bisantrene analogues were synthesized, bearing one or two 4,5-dihydro-1H imidazol-2-yl hydrazone side chains at positions 1,4 or 9 of the anthracene ring system. A 10-azabioisostere was also prepared. The position of substituents in structurally isomeric drugs modulates topoisomerase II poisoning and specificity, along with cytotoxicity. PMID- 9871639 TI - Synthesis and biological activity of glutamic acid derivatives. AB - In order to develop new specific glutamate analogues at metabotropic glutamate receptors, Diels-Alder, 1-4 ionic and radical reactions were performed starting from (2S)-4-methyleneglutamic acid. Preliminary pharmacological evaluation by measuring IP accumulation using rat forebrain synaptoneurosomes has shown that (2S)-4-(2-phthalimidoethyl)glutamic acid (3a), (2S)-4-(4 phthalimidobutyl)glutamic acid (3b) and 1-[(S)-2-amino-2-carboxyethyl]-3,4 dimethylcyclohex-3-ene-1-carbox ylic acid (8) presented moderate antagonist activities. PMID- 9871640 TI - S-aryl cysteine S,S-dioxides as inhibitors of mammalian kynureninase. AB - A series of 2-amino-S-aryl cysteine S,S-dioxides have been synthesised and shown to inhibit kynureninase an important enzyme in the biosynthesis of the known excitotoxic moiety quinolinic acid. The most potent of these, 2-amino-5-methyl-S phenyl cysteine S,S-dioxide 6d, inhibits interferon-gamma induced synthesis of quinolinic acid in human macrophages. PMID- 9871641 TI - Carbocyclic frenolicin analogues: novel anticoccidial agents. AB - Carbocyclic analogues of the antibacterial natural product frenolicin B have been synthesised. These analogues were active against parasitic protozoa of the genus Eimeria and represent a new series of anticoccidial agents. The synthesis of simplified analogues helped to define a possible pharmacophore for frenolicin. PMID- 9871642 TI - Alkylation of succinates: synthesis of Ro 32-3555. AB - A short synthesis, based on a succinate acylation-alkylation-decarboxylation approach, of the clinical compound Ro 32-3555 is reported. The nature of the selectivity in the mono-alkylation of succinates was examined under varying enolization conditions and in the presence of chaotropic additives. PMID- 9871643 TI - Synthesis and immunosuppressive activity of ruthenium complexes. AB - The syntheses and immunosuppressive activity of ruthenium complexes are described. One of the complexes (1a) was shown to be a potent inhibitor of human T-lymphocyte proliferation with an IC50 of 5 nM. The activity of these complexes compares favorably to the well known immunosuppressants Cyclosporin A and Rapamycin. PMID- 9871644 TI - A novel and practical route to A-ring enyne synthon for 1 alpha,25 dihydroxyvitamin D3 analogs: synthesis of A-ring diastereomers of 1 alpha,25 dihydroxyvitamin D2 and 3-methyl-1,25-dihydroxyvitamin D3. AB - A novel and practical route to the A-ring enyne synthon (2), which can be versatile for a variety of A-ring analogs of 1 alpha,25-dihydroxyvitamin D3 (1), was developed. This novel method led to an improved synthesis of the A-ring diastereomers of 1, the compounds 13-15, and synthesis of the new analogs, 2 methyl-1,25-dihydroxyvitamin D3 (4) with its all possible diastereomers. The biological evaluation of the 2-methyl analogs showed the alpha alpha beta-isomer to be more potent than 1. PMID- 9871645 TI - Enzymatic alpha(2-3)sialylation of non-natural type-I (Lewisc) disaccharides with recombinant sialyl-transferase. AB - Recombinant alpha(2-3)sialyl-transferase from rat liver is used to sialylate a series of type-I (Lewisc) disaccharides on a preparative scale. The enzyme tolerates a broad array of N-acetyl replacements of the N-glucosamine subunit ranging from small and large lipophilic groups to charged and heterocyclic amides. PMID- 9871646 TI - Enzymatic fucosylations of non-natural sialylated type-I trisaccharides with recombinant fucosyl-transferase-III. AB - Recombinant fucosyl-transferase-III (Lewis type enzyme) is used to prepare a series of non-natural sialyl-Lewis derivatives on a preparative scale. The enzyme tolerates a wide range of acceptors which have the natural N-acetyl group of the glucosamine moiety replaced by substituted aromatic and heteroaromatic amides. PMID- 9871647 TI - Alanyl-PNA homoduplex: A-T pairing with the N7-regioisomer of adenine. AB - The N7-regioisomer of adeninyl alanine can be used as a building block for the synthesis of alanyl-PNA. By changing the nucleobase connectivity from N9 to N7, pairing with the Hoogsteen side is no longer possible and the order of donor/acceptor positions at the Watson-Crick side is reversed. This influences the pairing selectivity but not the stability of alanyl-PNA self-pairing complexes, as shown by UV and CD spectroscopy. PMID- 9871648 TI - Remarkable tethering effect on DNA cleavage of propargylic sulfone conjugates with intercalating moieties. AB - A number of novel propargylic sulfone conjugates 3 and 4 with intercalating moieties were synthesized and evaluated for DNA cleavage activity through nucleic base alkylation. A remarkable enhancement in DNA cleaving potency was observed with those conjugates 3 possessing a suitable spacer, a right attachment point at the aromatic ring, and a good intercalator. PMID- 9871649 TI - Quaternary substituted PDE4 inhibitors I: the synthesis and in vitro evaluation of a novel series of oxindoles. AB - The following letter presents the synthesis and in vitro evaluation of a novel quaternary substituted series of phosphodiesterase type (IV) (PDE4) inhibitors. The compounds represent conformationally constrained analogues of the Celltech PDE IV inhibitor, CDP 840. Examples with sub-micromolar IC50's for PDE4 inhibition are reported. PMID- 9871650 TI - Synthesis and immobilization of ceramide analogs on silica particles. AB - Ceramides are the major lipid components of the stratum corneum, the major permeability barrier of the skin. Here we report a chemical synthesis of ceramide analogs covalently bonded on the silica particles, that can be used to predict the skin permeability of chemicals via HPLC methods. PMID- 9871651 TI - Synthesis and pharmacological evaluation of new flosulide analogues, synthesized from natural safrole. AB - Four new aryl-sulfonamide derivatives (3a, 4a, 5a-b), having methylenedioxy group attached to phenyl ring, were prepared from natural safrole and evaluated as anti inflammatory agents. The N-methylsulfonamide 3a and corresponding retrosulfonamide derivative 5a were more active than standards indomethacin and nimesulide, at the same molar concentration, in carrageenan-induced pleurisy assay. PMID- 9871652 TI - New taxanes as highly efficient reversal agents for multidrug resistance in cancer cells. AB - New non-cytotoxic taxanes synthesized from 10-deacetylbaccatin III and special hydrophobic acylating agents show remarkable MDR reversal activity (< or = 99.8%) against drug-resistant human breast cancer cells when co-administered with paclitaxel or doxorubicin. This activity is ascribed to the highly efficient blocking of P-glycoprotein efflux by these new taxanes. PMID- 9871653 TI - Synthesis of a non-nucleoside reverse transcriptase inhibitor in the alkenyldiarylmethane (ADAM) series with optimized potency and therapeutic index. AB - A novel alkenyldiarylmethane (ADAM) analog has been synthesized with enhanced potency as an anti-HIV agent. The new compound (ADAM II) inhibits the cytopathic effect of HIV-1RF in CEM-SS cells with an EC50 of 13 nM, while it shows cytotoxicity with a CC50 of 31.6 microM, providing a therapeutic index of 2430. ADAM II is a non-nucleoside reverse transcriptase inhibitor, displaying an IC50 of 0.3 microM with poly(rC) oligo(dG) as the template/primer. PMID- 9871654 TI - N-(2-(4-hydroxyphenyl)ethyl)-4-chlorocinnamide: a novel antagonist at the 1A/2B NMDA receptor subtype. AB - A series of N-(2-phenethyl)cinnamides was synthesized and assayed for antagonism at three N-methyl-D-asparate (NMDA) receptor subtypes (NR1A/2A-C). N-(2-(4 hydroxyphenyl)ethyl)-4-chlorocinnamide (6) was identified as a highly potent and selective antagonist of the NR1A/2B subtype. PMID- 9871655 TI - Stereochemistry of the reduction of 24-ethyldesmosterol to sitosterol in tissue cultures of Oryza sativa. AB - Feeding of [26-13C]- and [27-13C]-24-ethyldesmosterols to cultured cells of Oryza sativa followed by 13C-NMR analysis of the biosynthesized sitosterol revealed that the reduction of 24(25)-double bond proceeds with an anti-addition of hydrogen atoms, thus the E-methyl group of the olefinic precursor becomes the pro S-methyl on C-25 of sitosterol. PMID- 9871656 TI - Selective inhibition of the chymotrypsin-like activity of the 20S proteasome by 5 methoxy-1-indanone dipeptide benzamides. AB - Potent inhibitors of the 20S proteasome that contain a novel indanone head group coupled to di and tripeptides are described. These compounds are the first proteasome inhibitors have demonstrated high selectivity for the chymotrypsin like activity of the 20S proteasome. PMID- 9871657 TI - Immunomodulatory activity of thunberginol A and related compounds isolated from Hydrangeae Dulcis Folium on splenocyte proliferation activated by mitogens. AB - We investigated the immunomodulatory effects of antiallergic constituents from Hydrangeae Dulcis Folium, the processed leaves of Hydrangea macrophylla SERINGE var. thunbergii MAKINO, on splenocyte proliferation in mice. Thunberginol A and hydrangenol significantly suppressed T lymphocyte proliferation induced by concanavalin A. Thunberginol A also suppressed B lymphocyte proliferation induced by lipopolysaccharide, but other constituents induced significant increases. These inhibitory effects of thunberginol A on splenocyte proliferation seemed to contribute to the suppressive effect on type IV allergy. PMID- 9871658 TI - Methyloxime-substituted aminopyrrolidine: a new surrogate for 7-basic group of quinolone. AB - Novel fluoroquinolones containing oxime functionalized aminopyrrolidines have been synthesized. They were found to possess potent antibacterial activities against both Gram-negative and Gram-positive organisms, including methicillin resistant Staphylococcus aureus (MRSA). Among these compounds, LB20277 (compound 12) showed the most favorable in vivo efficacy and pharmacokinetic profile in animals. Based on these promising results, LB20277 was selected as a candidate for further evaluation. PMID- 9871659 TI - Synthesis and ESR study of new dihydroxamic acid siderophores S as scavengers of hydroxyl radicals. AB - Five new dihydroxamic acid ligands (L) (8, 10a, 10b, 10c and 13) have been synthesised and characterised as potential chelating agents for iron (Fe3+). The log stability constants of Fe2L3 and FeL+ from Fe3+ and L2- have been estimated to be log beta = 61.96 and log beta 1 = 22.8 respectively. The ability of these compounds to scavenge hydroxyl radicals (oOH) responsible for cell damage have been studied by esr spectroscopy. PMID- 9871660 TI - An alternative synthesis of 4,4-dimethyl-5 alpha-cholesta-8,14,24-trien-3 beta ol, an intermediate in sterol biosynthesis and a reported activator of meiosis and of nuclear orphan receptor LXR alpha. AB - 4,4-dimethyl-5 alpha-cholesta-8,14,24-trien-3 beta-ol, a sterol of current biological interest, has been synthesized in six steps from 3 beta-acetoxy-4,4 dimethyl-5 alpha-cholest-8(14)-en-15-one. PMID- 9871661 TI - Homologated aza analogs of arabinose as antimycobacterial agents. AB - A series of hydrolytically-stable aza analogs of arabinofuranose was prepared and evaluated against Mycobacterium tuberculosis and M. avium. The compounds were designed to mimic the putative arabinose donor involved in biogenesis of the essential cell wall polysaccharide, arabinogalactan. Though most compounds displayed little activity in cell culture, one compound showed significant activity in infected macrophage models. PMID- 9871662 TI - Trans-4-methyl-3-imidazoyl pyrrolidine as a potent, highly selective histamine H3 receptor agonist in vivo. AB - Extensive structural modification of immepyr (+)-2 led to the discovery of trans 4-methyl-3-imidazoyl pyrrolidine (+/-)-3a as a potent and highly selective H3 agonist. The pyrroline (+/-)-3a was resolved, and its (+) enantiomer, Sch 50971 [(+)-3a], showed a greater separation of H3 and H1 activities in vivo (H3/H1 ratio >> 330) than (R)-alpha-methylhistamine (+)-1 (H3/H1 ratio = 17), the standard H3 agonist. PMID- 9871664 TI - A practical synthesis of 3-[(1R)-1-t-butyldimethylsilyloxyethyl]-4-[(2R)- 4-halo 3-oxo-2-butyl]azetidinone, a versatile intermediate for carbapenem antibiotics. AB - A key intermediate for 2-(functionalized-methyl)-1-beta-methylcarbapenems, 3 [(1R)-1-t-butyldimethylsilyloxyethyl]-4-[(2R)- 4-halo-3-oxo-2-butyl]azetidinone, was prepared efficiently from a commercially available carboxylic acid in 3 steps. PMID- 9871663 TI - Synthesis and structure-activity relationships of pyridine-modified analogs of 3 [2-((S)-pyrrolidinyl)methoxy]pyridine, A-84543, a potent nicotinic acetylcholine receptor agonist. AB - Analogs of 3-[2-((S)-pyrrolidinyl)methoxy]pyridine, (A-84543, 1) with 2-, 4-, 5-, and 6-substituents on the pyridine ring were synthesized. These analogs exhibited Ki values ranging from 0.15 to > 9,000 nM when tested in vitro for neuronal nicotinic acetylcholine receptor binding activity. Assessment of functional activity at subtypes of neuronal nicotinic acetylcholine receptors indicates that pyridine substitution can have a profound effect on efficacy at these subtypes, and several subtype-selective agonists and antagonists have been identified. PMID- 9871665 TI - Synthesis of constrained alpha-amino acid derivatives via ring-closing olefin metathesis. AB - Five and seven membered constrained alpha-amino acid derivatives were synthesized using ring-closing metathesis reaction as a key step. PMID- 9871666 TI - Design and synthesis of a biotinylated dopamine transporter ligand for the purification and labeling of dopaminergic neurons. AB - The design and synthesis of a new tool for labeling and purification of dopaminergic neurons is described. PMID- 9871668 TI - A paclitaxel analogue with a 2(3-->20)abeotaxane skeleton: synthesis and biological evaluation. AB - A paclitaxel analogue having an unusual tricyclic [9.3.1.1] hexadecane skeleton was synthesized from deaminoacyltaxine A, a 2(3-->20) abeotaxane isolated in considerable amounts from the leaves of Taxus baccata L. In preliminary studies, this compound showed a much lower cytotoxicity than paclitaxel. PMID- 9871669 TI - Syntheses of new modified Phe-Pro peptides. Use of proline replacements in potential HIV inhibitors. AB - The syntheses consisting of replacement of proline amino acid by a 3 pyrrolidinone ring in Phe-Pro analogues are described. Preliminary anti-HIV studies demonstrated the potential activity of this new class of compounds. PMID- 9871667 TI - Synthesis of LIAZAL, a retinoic acid metabolism blocking agent (RAMBA) with potential clinical applications in oncology and dermatology. AB - The synthesis of LIAZAL (compound 9, R085246) is described. LIAZAL inhibits all trans-retinoic acid metabolism and thereby exerts retinoid-like effects in vivo. PMID- 9871671 TI - Rapid hydrolysis of amides under physiological conditions: influence of the microenvironment on the stability of the amide bond. AB - A new class of bicyclic carboxyamides 1a-9a differing with respect to substitution patterns and exo-endo geometry has been synthesized. These amides are characterized by a structure-dependent unusual rapid hydrolysis rate at physiological conditions. The corresponding bicyclic anhydrides might be used as tools for masking and modifying therapeutic agents containing amine functionalities. PMID- 9871670 TI - Synthesis and structure-activity relationships of CP-122,721, a second-generation NK-1 receptor antagonist. AB - The synthesis and SAR of benzylamine side chain analogs of the NK-1 receptor antagonist CP-99,994 are described. The 5-trifluoromethoxy analog, CP-122,721, shows superior in vivo blockade of NK-1 receptor mediated responses. PMID- 9871672 TI - Inhibitors of acyl-CoA:cholesterol O-acyltransferase (ACAT) as hypocholesterolemic agents: synthesis and structure-activity relationships of novel series of sulfonamides, acylphosphonamides and acylphosphoramidates. AB - Sulfoacetic acid, phosphoramidate, and phosphoramide analogs of the ACAT inhibitors, CI-999 and CI-1011 were synthesized. The structure-activity relationships of these compounds as ACAT inhibitors are described. PMID- 9871673 TI - New generation dopaminergic agents. 2. Discovery of 3-OH-phenoxyethylamine and 3 OH-N1-phenylpiperazine dopaminergic templates. AB - Described in this report is a systematic study which led to the identification of two new dopamine D2 partial agonists (5 and 17). Phenols 5 and 17 represent prototypes of two new classes of D2 partial agonists as well as templates for the future design of novel dopaminergic agents. PMID- 9871674 TI - Rational design of boropeptide thrombin inhibitors: beta, beta-dialkyl phenethylglycine P2 analogs of DuP 714 with greater selectivity over complement factor I and an improved safety profile. AB - The potent boropeptide thrombin inhibitor DuP 714 caused side effects in laboratory animals that appear to be related to its ability to inhibit complement factor I, thereby activating the complement cascade. Using X-ray crystal structure information, we have designed compounds that have greater selectivity for thrombin over factor I and that have reduced tendency to produce these side effects. PMID- 9871675 TI - Structure-activity relationships (SAR) of some tetracyclic heterocycles related to the immunosuppressive agent Brequinar Sodium. AB - The structure-activity relationships of some tetracyclic heterocycles related to Brequinar were explored. Activities as inhibitors of dihydroorotate dehydrogenase and the mixed lymphocyte reaction are related to ring system, heteroatom placement, and pendant ring substitution. PMID- 9871677 TI - Carboxy-substituted 2-azetidinones as cholesterol absorption inhibitors. AB - Metabolism initiated SAR studies led to the discovery of a new class of potent 2 azetidinone cholesterol absorption inhibitors. These studies found that a heteroatom at the para position of the C-4 phenyl ring is not a requirement for cholesterol absorption inhibition as was suggested by earlier findings. Substitution of Ph-linker-COOR for PhOMe at the C-4 position enhanced cholesterol absorption inhibition. PMID- 9871676 TI - Sugar-substituted 2-azetidinone cholesterol absorption inhibitors: enhanced potency by modification of the sugar. AB - A glucuronide conjugate of the potent 2-azetidinone cholesterol absorption inhibitor Sch 58235 was synthesized to confirm the structure of a metabolite isolated from in vivo sources. A series of 2-azetidinone glycosides was prepared via Schmidt trichloroimidate methodology. Enhanced cholesterol absorption inhibition was achieved by modification of the sugar moiety. PMID- 9871678 TI - Inhibition of thermolysin with nitrone-bearing substrate analogs: a new type of thermolysin inhibitors. AB - Nitrones are utilized as the active site zinc coordinating functionality in the design of inhibitors for thermolysin. This new type of thermolysin inhibitors are as potent as the existing inhibitors bearing a carboxylate or hydroxamate zinc ligating moiety. PMID- 9871679 TI - Immunosuppressant activity in human beta-casein fragment analogs. AB - Human beta-casein fragment (54-59) having the amino acid sequence Val-Glu-Pro-Ile Pro-Tyr, has been shown potent immunostimulant activity. Several analogs of this hexapeptide have been synthesized with modification in the N-terminal region and tested for their immunomodulatory activity. Interestingly, two hexapeptides have shown significant immunosuppressant activity. PMID- 9871680 TI - Potent and selective inhibitors of the proteasome: dipeptidyl boronic acids. AB - Potent and selective dipeptidyl boronic acid proteasome inhibitors are described. As compared to peptidyl aldehyde compounds, boronic acids in this series display dramatically enhanced potency. Compounds such as 15 are promising new therapeutics for treatment of cancer and inflammatory diseases. PMID- 9871681 TI - Inhibitory effect and action mechanism of sesquiterpenes from Zedoariae Rhizoma on D-galactosamine/lipopolysaccharide-induced liver injury. AB - Hepatoprotective sesquiterpenes were isolated from the aqueous acetone extract of Zedoariae Rhizoma, the rhizome of Curcuma zedoaria ROSCOE (Zingiberaceae). Principal sesquiterpenes, furanodiene, germacrone, curdione, neocurdione, curcumenol, isocurcumenol, aerugidiol, zedoarondiol, and curcumenone and curcumin were found to show potent protective effect on D-galactosamine (D GalN)/lipopolysaccharide (LPS)-induced acute liver injury in mice. Plausible action mechanisms for their hepatoprotective activity were clarified on the basis of the inhibitory effect on D-GalN-induced cytotoxicity in primary cultured rat hepatocytes, LPS-induced NO production in cultured mouse peritoneal macrophages, and D-GalN/tumor necrosis factor-alpha (TNF-alpha)-induced liver injury in mice. PMID- 9871682 TI - Naphthalenebis[alpha,alpha-difluoromethylenephosphonates] as potent inhibitors of protein tyrosine phosphatases. AB - A series of naphthalenebis(difluoromethylenephosphonates) were prepared and compared to their monosubstituted counterparts as inhibitors of the protein phosphatases, PTP1B, CD45 and PP2A. In general, the bissubstituted compounds were better inhibitors than the mono derivatives and some of these are among the most potent, nonpeptidyl inhibitors of protein tyrosine phosphatases, PTP1B and CD45, reported to date. PMID- 9871683 TI - Thiophilic ring-opening and rearrangement reactions of epoxyketone natural products. AB - Thiol additions to the highly functionalized core structures of aranorosin- and manumycin-type antibiotics reveal the general reactivity patterns of epoxyketone natural products. Rapid hemiacetal and hydrate formations decrease the reactivity of the epoxyketone moiety in aqueous media toward the cellular scavenger glutathione, and secondary 1,2-shift, elimination, aromatization and intramolecular aldol reactions provide novel reaction pathways. In a hydrophobic environment, the thiol-capture function of the ketone moiety facilitates electrophilic attack. PMID- 9871684 TI - Synthesis of novel cyclic protease inhibitors using Grubbs olefin metathesis. AB - The unusual amino acid bishomoallylglycine was synthesized and used to form cyclic P3-P1 tripeptide inhibitors via a Grubbs olefin metathesis method. These compounds show micro- to nanomolar inhibition of Rhizopus chinensis pepsin and represent a new class of simplified aspartic protease inhibitors lacking P' residues. PMID- 9871685 TI - Single bead IR monitoring of a novel benzimidazole synthesis. AB - A novel and efficient solid-phase synthesis of benzimidazoles and the monitoring of all conversions in this synthesis are reported. PMID- 9871686 TI - Design and synthesis of monocyclic beta-lactams as mechanism-based inhibitors of human cytomegalovirus protease. AB - Mechanism based inhibitors of HCMV protease have been designed based on the monocyclic beta-lactam nucleus, which have been shown to acylate the viral enzyme in a time dependent manner. SAR in a series of monocyclic beta-lactam N-ureas, has defined the size and relative stereochemistry of the C-3 substituent producing a low micromolar inhibitor 17b with good aqueous stability and selectivity over the mammalian serine proteases. PMID- 9871687 TI - A new strong inhibitor of beta-mannosidase. AB - N-phenyl-carbamate of D-mannonohydroxymolactone (I) was synthesized from mannose and was shown to be the best competitive inhibitor of beta-mannosidase so far reported (Ki = 25 nM). PMID- 9871688 TI - Inhibitors of the chymotrypsin-like activity of proteasome based on di- and tri peptidyl alpha-keto aldehydes (glyoxals). AB - A series of peptidyl alpha-keto aldehydes (glyoxals) have been synthesised as putative inhibitors of the chymotryptic-like activity of proteasome. The most potent peptides, Cbz-Leu-Leu-Tyr-COCHO and Bz-Leu-Leu-Leu-COCHO, function as slow binding reversible inhibitors, exhibiting final Ki values of approximately 3.0 nM. These are among the lowest values so far reported for (tri)peptide-based aldehyde-related inhibitors. PMID- 9871689 TI - Synthesis of alpha-aryl N-adamant-1-yl nitrones and using them for spin trapping of hydroxyl radicals. AB - alpha-Aryl N-adamant-1-yl nitrones were synthesized and evaluated with respect to the stability of the hydroxyl radical adduct. The polarity and water solubility of nitrones were altered with changing the alpha-aryl groups. Introduction of adamantane ring instead of tert-butyl group resulted in a reasonable good stability of hydroxyl radical adduct for biological measurements. PMID- 9871690 TI - Synthesis of Dolastatin 15 mimetic peptoids. AB - Eight peptoids have been synthesized as peptidomimetics of the cytostatic Dolastatin 15, a depsipeptide isolated from the Indian sea hare Dolabella auricularia. The compounds have been tested against several human cancer cell lines and did not show any cytostatic properties. PMID- 9871691 TI - Heteroatom-substitution as a strategy for increasing the potency of competitive NMDA antagonists. AB - We report the synthesis and characterization of compounds that are competitive NMDA receptor antagonists. Significant increases in affinity and potency were obtained by incorporation of a heteroatom into the substructure of the tetrazole substituted amino acid LY233053. PMID- 9871692 TI - Synthesis and 5 alpha-reductase inhibitory activity of 8-substituted benzo[f]quinolinones derived from palladium mediated coupling reactions. AB - Benzoquinolinones have been shown to be potent, selective inhibitors of the Type I 5 alpha-reductase enzyme, which is responsible for the production of dihydrotestosterone from testosterone localized in the scalp. In an effort to identify compounds that demonstrate inhibition of both 5 alpha-reductase isozymes, we have employed 8-bromobenzoquinolinone as an advanced intermediate for participation in a variety of palladium mediated carbon-carbon bond forming reactions. By varying the 8-substituent it is possible to alter the selectivity profile of the series. PMID- 9871693 TI - Quaternary substituted PDE IV inhibitors II: the synthesis and in vitro evaluation of a novel series of gamma-lactams. AB - This communication describes the synthesis and in vitro evaluation of a novel potent series of phosphodiesterase type (IV) (PDE IV) inhibitors. Several of the quaternary substituted lactams presented possess low nanomolar IC50's for PDE IV inhibition. PMID- 9871694 TI - Synthesis of L-enantiomers of 4'-thioarabinofuranosyl pyrimidine nucleosides. AB - L-Enantiomers of 4'-thioarabinofuranosyl pyrimidine nucleosides were synthesized from D-xylose. Methyl 2,3,5-tri-O-benzyl-D-xylofuranoside 6 was converted to the corresponding xylitol 7, which was treated with MsCl and then Na2S to give 1,4 anhydro-L-4-thioarabitol 8. As previously reported, Pummerer rearrangement of 8 followed by glycosylation with a silylated thymine and N4-acetylcytosine derivative and deprotection gave the corresponding alpha- and beta-L-4' thioarabinofuranosyl pyrimidine nucleosides. PMID- 9871695 TI - The design and synthesis of inhibitors of the cysteinyl protease, Der p I. AB - Prototype irreversible inhibitors of the cysteinyl protease Der p I were designed, synthesised and evaluated in vitro. Candidates were designed using a modular approach, whereby a peptide sequence was appended with known thiophilic moieties. This hinged on utilizing peptide sequences from substrate specificity data compiled using proprietary RAPiD technology. PMID- 9871696 TI - Novel euglycemic and hypolipidemic agents: Part-2. Antioxidant moiety as structural motif. AB - Several thiazolidinediones having antioxidant moities in their structural motif have been synthesised and evaluated for their euglycemic and hypolipidemic activities. A few of them have been found to be superior to troglitazone. PMID- 9871697 TI - Stability of acetal and non acetal-type analogs of artemisinin in simulated stomach acid. AB - A series of non acetal-type analogs of artemisinin containing C-C bond at position-12 have been found to be 15-22 times more stable than acetal(C-O)-type prodrugs of artemisinin in simulated stomach acid. PMID- 9871698 TI - Complexation of okadaic acid: a preliminary study. AB - A complexation study was carried out with okadaic acid (OA) and the univalent metal ions Li+, Na+ and K+, and the divalent metal ions Ca2+ and Mg2+. K+ binding was observed identical with a complex obtained from the natural source (OAC). The pharmacological trials demonstrated that this cation has a very important influence on the pharmacological activity of okadaic acid. PMID- 9871699 TI - Synthesis of DNA analogues with novel carboxamidomethyl phosphonamide and glycinamide internucleoside linkages. AB - Thymidine oligonucleotide analogues with phosphodiester bonds fully substituted by carboxamidomethyl phosphonamide, or glycinamide linkages were synthesized on a solid support, and their hybridization properties toward DNA and RNA targets were determined by Tm analysis. PMID- 9871701 TI - Mechanism of 17 alpha-hydroxylase/17,20-lyase--an initial geometric perspective for the lyase of the C(17)-C(20) bond of C21 steroids. AB - Using the novel 'substrate-heme complex' approach, the mechanism of 17 alpha Hydroxylase/17,20-Lyase (P-450(17) alpha), in particular the lyase of the C(17) C(20) bond, is considered from a geometric perspective. The results of the study appear to suggest that the final oxidative step in the lyase of the C(17)-C(20) bond involves the use of a ferroxy attacking species as opposed to peroxy or a mixture of ferroxy and peroxy, an observation which is consistent with results previously obtained with Aromatase. PMID- 9871700 TI - Design of new inhibitors for cdc2 kinase based on a multiple pseudosubstrate structure. AB - New inhibitors have been designed for cdc2 kinase based on a multiple pseudosubstrate structure. The new inhibitors have three different structural components: 3,4-bis(indol-3-yl)maleimide, Ac-Cys-(Ser)-Pro-Lys-Lys-NHMe, and ethyloxy group between the two components. Inhibitory activities toward cdc2 and other protein kinases were investigated, and the compound (21) with Ac-Cys-Pro Lys-Lys-NHMe connected with the triethylene glycol spacer exhibited the most potent inhibition with relatively high selectivity. PMID- 9871702 TI - On the use of volume maps in the conformational analysis of vitamin D analogs. AB - Dot maps used to represent the calculated conformations of the side chain of vitamin D derivatives are improved by replacing dots by coloured balls to create volume maps and to identify particular energy windows. Two procedures to define a relative activity volume, based on a pair of analogs with distinctly different biological activity and conformational behaviour, are presented. PMID- 9871703 TI - PF1022A--a novel anthelmintic cyclooctadepsipeptide. Modification and exchange of the N-methyl leucine residues. AB - The first structure-activity relationships of the anthelmintic cyclooctadepsipeptide PF1022A have been established via a systematic exchange of the leucine residues by a series of related N-alkylated amino acids. The data presented strongly suggest that (L)-N-methyl-leucine is crucial for high in vivo activity. PMID- 9871705 TI - Anti-HIV pronucleotides: decomposition pathways and correlation with biological activities. AB - The purpose of the present study was to compare the decomposition pathways in CEM cell extracts of various phenyl phosphoramidate derivatives of AZT. In addition, the structures of their metabolites were identified. Correlations with their anti HIV activities in a thymidine kinase deficient (TK-) CEM cell line have been established with a rationale of designing phosphoramidate pronucleotides capable of delivering intracellularly their respective 5'-nucleoside monophosphate derivatives. PMID- 9871704 TI - Design and synthesis of a new type of non steroidal human aromatase inhibitors. AB - The structure-activity relationship study of one of recently described aromatase inhibitors, compound 1 (MR20814), allowed us to design some related derivatives as potential new inhibitors. Among those we synthesized, chlorophenylpyridylmethylenetetrahydroindolizinone 5 (MR20492) exhibited in vitro a ten-fold higher inhibition of the enzyme (IC50 = 0.2 +/- 0.0 microM and Ki = 10.3 +/- 3.3 nM). PMID- 9871706 TI - Potent antimitotic and cell growth inhibitory properties of substituted chalcones. AB - A series of substituted chalcones was synthesised and screened for cytotoxic activity against the K562 human leukaemia cell line. (E)-3-(3"-Hydroxy-4" methoxyphenyl)-2-methyl-1-(3',4',5'- trimethoxyphenyl)-prop-2-en-1-one [IC50 (K562) 0.21 nM] was found to be the most active. A relationship between the conformation and cytotoxicity of the chalcones is discussed. PMID- 9871707 TI - A thiophosphate analog of dimyristoylphosphatidyl-inositol-4-phosphate is a substrate for mammalian phosphoinositide-specific phospholipase C. AB - 1,2-Dimyristoyloxypropane-3-thiophosphate(rac-1-myo-inositol-4- phosphate), a thiophosphate analog of dimyristoyl phosphatidylinositol-4-phosphate was synthesized as a substrate for mammalian phosphoinositide-specific phospholipase C. Its activity with delta(1-132)-PI-PLC-delta 1 (a deletion mutant with the N terminal pleckstrin homology domain removed) was studied in sonicated dispersions, with and without added Triton X-100. It had an initial activity of about 30 mumol min-1 mg-1, which rapidly decreased due to substrate depletion in the vesicle or micelle. The slower rate of hydrolysis appeared limited by enzyme hopping or exchange of substrate between vesicles or micelles, which was more rapid in the presence of detergent. PMID- 9871708 TI - Synthesis of 2-deoxy-2-fluoro-phosphatidylinositol-4,5-bisphosphate and analogues: probes and modulators of the mammalian PI-PLCS. AB - An approach to synthesis of 2-modified phosphatidylinositol-4,5-bisphosphates, which are substrate analogues useful as probes and modulators of the PI-PLC enzyme family, is described and illustrated for the dibutyl-2-deoxy-2-fluoro analogue, a probe designed for delineating substrate and PI-PLC interactions by X ray crystallography. PMID- 9871709 TI - The synthesis and evaluation of benzannelated-azatoxins: the benzazatoxins. AB - The synthesis and evaluation of azatoxin congeners possessing annealed aromatic frameworks are described. The compounds were evaluated for their abilities to affect topoisomerase II inhibition through the stabilization of "cleavable complex" and for the inhibition of tubulin polymerization using purified bovine brain tubulin. PMID- 9871710 TI - Enhanced potency of perfluorinated thalidomide derivatives for inhibition of LPS induced tumor necrosis factor-alpha production is associated with a change of mechanism of action. AB - Perfluorination of phthalimides leads to dramatically increased potency as inhibitors of TNF-alpha production. We examined the enantiodependence for several tetrafluorophthalimides and alpha-methylthalidomide, 3. Only 3 exhibited strikingly enantiodependent activity. The key structural determinant for the enhanced activity is the tetrafluorophthaloyl group, which confers enhanced potency and a change in the mechanism of inhibition. PMID- 9871711 TI - The synthesis of symmetrical and unsymmetrical P1/P1' cyclic ureas as HIV protease inhibitors. AB - Cyclic urea SD146, a potent HIV protease inhibitor bearing a flat resistance profile, possessed poor solubility and bioavailability, which precluded further development of the compound. In an effort to improve upon the pharmacokinetic profile of the compound, several analogs modified at the P1/P1' residues were prepared and evaluated. Several of those compounds displayed significant improvement of physical properties. PMID- 9871713 TI - Polymer supported bases in solution-phase synthesis. 2. A convenient method for N alkylation reactions of weakly acidic heterocycles. AB - A convenient solution-phase method for N-alkylation reactions of weakly acidic heterocycles employing polymer supported super base PBEMP is described. By using this method, multiple-step and chemoselective N-alkylation sequences can be carried out in a one-pot process. PMID- 9871712 TI - Prediction of the GTPase activities by using the semiempirical molecular orbital theory. AB - Assuming that substrate-assisted catalysis is the mechanism of GTP hydrolysis for ras p21 and other GTP-binding proteins, we used the PM3 semiempirical molecular orbital method to predict from the calculated reaction profiles of GTP hydrolysis reactions the changes in GTPase activities caused by mutations. We succeeded in making qualitative predictions for mutants. PMID- 9871714 TI - Anti-inflammatory/antiarthritic ketonic bisphosphonic acid esters. AB - Bisphosphonate ester 2 is an inhibitor of inflammation, but is devoid of antiarthritic effects. SAR studies on a series of related bisphosphonate esters resulted in compounds 6e, 6i, 6j, and 6m, which exhibited excellent inhibition of an arthritis model, in addition to potent anti-inflammatory effects. PMID- 9871715 TI - Synthesis of beta-difluorine-containing amino acids. AB - A convenient strategy was developed to prepare several beta-difluoroamino acids. As exemplified by the synthesis of 3,3-difluoro-L-homocysteine, 3,3-difluoro-L homoserine and 3,3-difluoro-L-methionine, the reaction sequence all started from L-isoascorbic acid. This approach holds potential to be extended to make other beta-difluorine-containing amino acids. PMID- 9871716 TI - Potent, selective benzenesulfonamide agonists of the human beta 3 adrenergic receptor. AB - A cloned human beta 3 adrenergic receptor assay was used to identify phenoxypropanolamine agonist 1. SAR studies led to the identification of benzenesulfonamide derivative 20, a 6.3 nM beta 3 agonist which shows 30-fold selectivity for beta 3 agonist activity over beta 1 and beta 2 receptor binding. Further refinement of this lead provided 4-bromo derivative 39, a subnanomolar agonist with 660-fold and 230-fold selectivity over beta 1 and beta 2, respectively. PMID- 9871717 TI - Discovery of L-755,507: a subnanomolar human beta 3 adrenergic receptor agonist. AB - A study of 4-acylaminobenzenesulfonamides in a cloned human beta 3 adrenergic receptor assay resulted in the discovery of n-hexylurea, L-755,507 (22). This 0.43 nM beta 3 agonist, which is > 440-fold selective over both beta 1 and beta 2 binding, is among the most potent human beta 3 agonists reported to date. PMID- 9871718 TI - An enantioselective fluorimetric assay for alcohol dehydrogenases using albumin catalyzed beta-elimination of umbelliferone. AB - 3-hydroxybutyl umbelliferyl ethers (R)-1 and (S)-1 are fluorogenic substrates for alcohol dehydrogenases. Their oxidation forms ketone 2, which undergoes beta elimination of umbelliferone under catalysis by bovine serum albumin, leading to a > 20-fold fluorescence increase at lambda em = 460 +/- 20 nm (lambda ex = 360 +/- 20 nm). Enantioselectivity is determined in two separate tests with each enantiomeric substrate. PMID- 9871719 TI - Serotonergic ergoline derivatives. AB - Novel classes of 13- and 14-tertbutyl-ergoline derivatives were prepared, and characterised in vitro for their affinity for adrenergic, dopaminergic and serotonergic binding sites. This study particularly examines the importance of the presence and the position of the tert-butyl group in conferring either significant 5-HT1A or 5-HT2 affinity and selectivity respectively. PMID- 9871720 TI - Synthesis of 14 C-radiolabeled oligonucleotides with a novel phosphoramidite reagent. AB - A novel solid-phase synthesis of 5'-radiolabeled oligonucleotides is described. The labeling reaction is carried out by the phosphoramidite method with the aid of [4,6-di-14C]-5'-dimethoxytritylthymidine building block 1. The feasibility of the method is demonstrated by preparation of 3'-phosphorylated dodecathymidylate phosphorothioate containing radiolabeled nucleoside at the 5'-terminus. PMID- 9871721 TI - Diversity of C-linked neoglycopeptides for the exploration of subsite-assisted carbohydrate binding interactions. AB - Diversity of alpha-galactose based C-linked neoglycopeptides (1b, 2b, 3c, 4d, and 5d) has been developed to explore the importance of subsite-assisted carbohydrate binding interactions. Deprotected C-linked neoglycopeptides (1b, 2b, 3c, 4d, and 5d) were synthesized and tested in competitive inhibition assays using a model enzyme-linked lectin (e.g., Maclura pomifera). Compound 2b, with two alpha galactoside units on the side chain of the lysine residue of the dipeptide backbone, exhibited a remarkable effect with a 2.82-fold increase in its inhibitory properties (IC50 1.48 mM) in comparison to 1b (IC50 4.18 mM). PMID- 9871722 TI - New acetylene based histamine H3 receptor antagonists derived from the marine natural product verongamine. AB - New histamine H3 receptor antagonists were developed using an acetylene moiety as a replacement for the amide-oxime functionality of verongamine 5. Optimization of receptor binding was performed by following aliphatic Topliss tree guidelines. These new H3 ligands demonstrate excellent blood-brain barrier penetration. PMID- 9871723 TI - Substrate specificity of human prolyl-4-hydroxylase. AB - Proline analogs (3-F, 3-Cl, 3-Br, 3,3-cyclopropyl, 3,3-methylene, 3-Me, and 4-Me) were synthesized, incorporated into CbzGlyPheXGlyOEt, and tested as substrate analogues/mechanistic probes for the human prolyl-4-hydroxylase catalyzed hydroxylation reaction. With the exception of the 3-fluoroproline containing peptide, none of these peptides were substrates for the enzyme. PMID- 9871724 TI - alpha-Mannosidase activity from antibody raised against a glucal antigen. AB - Sixty cell lines of monoclonal antibody were raised against a glucal hapten 1. Among them, Ab 405.4 showed alpha-mannosidase activity as kcat = 0.19/day (kcat/kuncat = 110,000). The chemical modification study and pH profile study of this antibody indicated that carboxyl group(s) in the antigen binding site involved in catalytic mechanism. PMID- 9871725 TI - Recognition of a non-standard base pair by thermostable DNA polymerases. AB - Examination of several commercially available thermostable DNA polymerases identifies 9 degrees N DNA polymerase as single enzyme that could incorporate two components of an expanded genetic alphabet, 2,4-diaminopyrimidine and xanthosine as deoxynucleoside triphosphate opposite their cognate base in a DNA template. PMID- 9871726 TI - A novel super-potent neurokinin A receptor antagonist containing dehydroalanine. AB - We report here the synthesis and preliminary pharmacological characterization of a novel Neurokinin A receptor antagonist. This molecule contains a dehydroalanine residue. It displays a high conformational rigidity and possesses very high activity. Its pharmacological properties as a neurokinin A receptor antagonist were assessed in in vitro experiments on rat vas deferens and were compared to those of Neuronorm and MEN10627. PMID- 9871727 TI - Malonyl alpha-mercaptoketones and alpha-mercaptoalcohols, a new class of matrix metalloproteinase inhibitors. AB - A novel series of matrix metalloproteinase (MMP) inhibitors is described. Incorporation of a terminal alpha-mercaptoketone or alpha-mercaptoalcohol in the zinc binding domain of a series of inhibitors led to compounds exhibiting low nanomolar activity against collagenase-1 (MMP-1), stromelysin (MMP-3), and gelatinase-B (MMP-9). PMID- 9871728 TI - The asymmetric synthesis and in vitro characterization of succinyl mercaptoalcohol and mercaptoketone inhibitors of matrix metalloproteinases. AB - A series of succinyl based mercaptoketones and diastereomeric mercaptoalcohols were prepared and evaluated in vitro as inhibitors of the matrix metalloproteinases collagenase-1 (MMP-1), stromelysin (MMP-3), and gelatinase-B (MMP-9). PMID- 9871729 TI - Synthesis and anti-inflammatory activity of chalcone derivatives. AB - Chalcones and their derivatives were synthesized and evaluated for their anti inflammatory activity. In vitro, chalcones 2, 4, 8, 10 and 13 inhibited degranulation and 5-lipoxygenase in human neutrophils, whereas 11 behaved as scavenger of superoxide. Only four compounds (4-7) inhibited cyclo-oxygenase-2 activity. The majority of these samples showed anti-inflammatory effects in the mouse air pouch model. PMID- 9871730 TI - Structure-activity relationships in 2-aminodiphenylsulfides against trypanothione reductase from Trypanosoma cruzi. AB - In order to establish structural elements responsible for inhibition of trypanothione reductase (TR) from Trypanosoma cruzi by 2-aminodiphenylsulfides, a series of dissymmetrical derivatives, corresponding to the replacement of one aromatic moiety by different amines, was synthesized. TR inhibition studies revealed the importance of the aromatic rings and of the amino groups in the side chains for potent inhibition. Quinonic moities were also introduced with the aim of acting as TR redox-cycling substrates. PMID- 9871731 TI - Antiinflammatory 2-benzyl-4-sulfonyl-4H-isoquinoline-1,3-diones: novel inhibitors of COX-2. AB - A series of 2-benzyl-4-sulfonyl-4H-isoquinoline-1,3-diones was prepared. Members of this series are potent and selective inhibitors of cyclooxygenase-2 (COX-2) in both microsomal and cellular assays. Two representatives demonstrated activity in the carrageenan-induced paw edema model in rats upon oral administration. PMID- 9871732 TI - Synthesis of [(2'S, 3'S)-bis(hydroxylmethyl)pyrrolidin-1-yl] purine and pyrimidine nucleosides as potential antiviral agents. AB - The enantiomerically pure synthesis of [(2'S, 3'S)-bis(hydroxymethyl)pyrrolidin-1 yl] thymine 17 and -adenine 20 was achieved via construction of the base on the 1 amino-pyrrolidine 15, and their anti-HSV-1 and -2, and anti-HIV-1 activities were evaluated. PMID- 9871733 TI - The formation of a covalent complex between a dipeptide ligand and the src SH2 domain. AB - The X-ray crystal structure of the src SH2 domain revealed the presence of a thiol residue (Cys 188) located proximal to the phosphotyrosine portion of a dipeptide ligand. An aldehyde bearing ligand (1) was designed to position an electrophilic carbonyl group in the vicinity of the thiol. X-ray crystallographic and NMR examination of the complex formed between (1) and the src SH2 domain revealed a hemithioacetal formed by addition of the thiol to the aldehyde group with an additional stabilizing hydrogen bond between the acetal hydroxyl and a backbone carbonyl. PMID- 9871734 TI - Synthesis of tools for target identification of the anti-apoptotic compound CGP 3466; Part I. AB - Immobilized compounds for BIAcore studies and affinity precipitation as well as a fluorescent-labeled compound were prepared in order to identify the molecular target of the anti-apoptotic, neurorescuing compound CGP 3466 (N-methyl-N propargyl-10-aminomethyl-dibenzo[b,f]oxepin). PMID- 9871735 TI - In vitro evaluation of synthetic heparin-like conjugates comprising different thrombin binding domains. AB - The syntheses of several heparin-like glycoconjugates (i.e., 16a-f) containing identical AT III binding domains (ABD) and spacers but different thrombin binding domains (TBDs) are described. Biological activities of conjugates 16a-f indicate that the thrombin inhibitory activity is mainly determined by the charge density of the TBD moiety. PMID- 9871736 TI - Direct synthesis of [DOTA-DPhe1]-octreotide and [DOTA-DPhe1,Tyr3]-octreotide (SMT487): two conjugates for systemic delivery of radiotherapeutical nuclides to somatostatin receptor positive tumors in man. AB - Direct attachment of unprotected DOTA (1,4,7,10-tetraazacyclododecane N',N",N"',N""-tetraacetic acid) to partially suitably protected octreotide or [Tyr3]-octreotide leads after deprotection to [DOTA-DPhe1]-octreotide (III) and [DOTA-DPhe1,Tyr3]-octreotide (IV). These DOTA-containing somatostatin analogs, when labeled with a radiotherapeutic nuclide, are useful as antitumor agents. The partially protected peptides are accessible via solid phase peptide synthesis (SPPS) followed by selective cleavage under mild acidic conditions from the resin. PMID- 9871737 TI - Design and synthesis of bio-isosteres of thymidine triphosphate. AB - This paper describes the design and synthesis of potential isosteres of triphosphates which should show enhanced metabolic stability and lipophilicity compared to triphosphates. The triphosphate isosteres were then linked to nucleosides and evaluated for their inhibitory activity against HIV infection. PMID- 9871738 TI - Syntheses of 1-deoxy-3-s-(1-thio-alpha-D-glucopyranosyl)-mannojirimycin and 1 deoxy-3-O-(5-thio-alpha-D-glucopyranosyl)-mannojirimycin as potential inhibitors of endo-alpha-D-mannosidase. AB - 1-Deoxy-3-S-(1-thio-alpha-D-glucopyranosyl)-mannojirimycin and 1-deoxy-3-O-(5 thio-alpha-D-glucopyranosyl) mannojirimycin were chemically synthesized as potential inhibitors of endo-alpha-D-mannosidase. PMID- 9871739 TI - High-throughput synthesis and direct screening for the discovery of novel hydrolytic metal complexes. AB - A 'combinatorial catalysis' strategy was utilized to both rapidly synthesized organometallic complexes and to screen them for catalytic activity in chemical reactions. The application of this strategy has yielded several metal-complexes that catalyze the hydrolysis of carboxylic acid esters efficiently. PMID- 9871740 TI - UV spectroscopy study on complexes of phosphonate ApA analogs with poly(U): promising step in prediction of oligonucleotide analog properties? AB - The isopolar nonisosteric phosphonate analogs of ApA differing in the position of extra methylene group introduced into the sugar-phosphate backbone, featuring both possible 2',5'- and 3',5'- pairs as well as their conformationally restricted congeners, were investigated for their ability to form complexes with polyU. The results may lead to the specification of candidates for synthesis of novel oligonucleotides. PMID- 9871741 TI - Synthesis and antibacterial activity of [6,5,5] and [6,6,5] tricyclic fused oxazolidinones. AB - A series of conformationally restricted, [6,5,5] and [6,6,5] tricyclic fused oxazolidinones were synthesized and tested for antibacterial activity. Several compounds in the trans-[6,5,5] series demonstrated potent in vitro and in vivo activity. This work provides valuable information regarding the preferred conformational orientation of the oxazolidinones at the binding site. PMID- 9871742 TI - Non-peptidic HIV protease inhibitors: C2-symmetry-based design of bis-sulfonamide dihydropyrones. AB - Potent, non-peptidic, dihydropyrone sulfonamide HIV protease inhibitors have been previously described. Crystallographic analysis of dihydropyrone sulfonamide inhibitor/HIV protease complexes suggested incorporation of a second, C2 symmetry related sulfonamide group. Selected bis-sulfonamide dihydropyrone analogues display high HIV protease inhibitory activity. PMID- 9871743 TI - Effect of side chain location in (2-aminoethyl)-aminomethyl-2-phenylquinolines as antitumor agents. AB - Three new derivatives of 2-phenylquinoline having an (2-aminoethyl)aminomethyl group in 7-, 6-, or 4'- (para position of 2-phenyl ring) positions of aromatic system have been prepared. The antitumor activity of these compounds together with 8- or 4- substituted isomers against the HeLa cell is in the order of 8- > 7 > 4- approximately 6- approximately 4'- substituted ones, which is almost in good agreement with that of DNA-binding ability evaluated by means of DNA titration of UV-VIS spectra, DNA melting experiment, and ethidium displacement assay. Two representative compounds (8- and 4- isomers) are confirmed to have an ability to intercalate into double stranded DNA by topoisomerase I superhelix unwinding assay. PMID- 9871744 TI - 1,2-disubstituted cyclohexane derived tripeptide aldehydes as novel selective thrombin inhibitors. AB - A series of tripeptide arginine aldehydes was synthesized by replacement of proline with 1,2-disubstituted cyclohexane derivatives in the sequence of D-MePhe Pro-Arg-H. Based on molecular modeling, further modification of the D-MePhe residue resulted in a potent and selective thrombin inhibitor. PMID- 9871745 TI - Quinolines as potent 5-lipoxygenase inhibitors: synthesis and biological profile of L-746,530. AB - Leukotriene biosynthesis inhibitors have potential as new therapeutic agents for asthma and inflammatory diseases. A series of novel substituted 2-cyanoquinolines have been synthesized and the structure activity relationships were evaluated with respect to their ability to inhibit the formation of leukotrienes via the 5 lipoxygenase enzyme. [1S,5R]-2-Cyano-4-(3-furyl)-7-?3-fluoro-5-[3-(3 alpha hydroxy-6,8-dioxabicyclo[3.2.1]-octanyl)]phenoxymethyl ?quinoline (L-746,530) 3 represents a distinct class of inhibitors and possesses in vitro and in vivo potency comparable or superior to naphthalenic analog (L-739,010) 2. PMID- 9871746 TI - 3-trifluoromethylcarbacephems: synthesis of broad spectrum antibacterial compounds. AB - The enhanced stability of the carbacephem nucleus over the corresponding cephalosporin nucleus has allowed the synthesis of 7-arylglycyl-3-trifluoromethyl carbacephems. These unique carbacephems possess broad spectrum activity and high stability to both plasmid and chromosomally mediated beta-lactamases. PMID- 9871747 TI - Anti-AIDS agents. 32. Synthesis and anti-HIV activity of betulin derivatives. AB - Eleven betulin derivatives were prepared and evaluated for anti-HIV activity in H9 lymphocytes. Compound 4 was found to be the most active with EC50 and TI values of 0.00066 microM and 21,515, respectively. PMID- 9871748 TI - An alternative synthesis of peptidyl alpha-keto-2-oxazolines. AB - Application of Wipf's protocol to generate alpha-keto-2-oxazolines is described. PMID- 9871749 TI - A depsipeptide fungal metabolite inhibitor of cholesteryl ester transfer protein. AB - The organic extract of the fermentation broth of a fungus was found to contain a depsipeptide SCH 58149 (1), containing three amino acids and a beta-hydroxy acid, by spectroscopic studies. The amino acids were phenyl alanine, alanine and leucine and the beta-hydroxy acid is 3-hydroxy-4-methyl octanoic acid. SCH 58149 exhibited weak activity against cholesterol ester transfer protein (CETP) with an IC50 of 50 microM. PMID- 9871750 TI - Estradiol-mimetic probes. Preparation of 17 alpha-(6-aminohexynyl)estradiol biotin, fluorescein and acridinium conjugates. AB - 3-O-tert-Butyldimethylsilyl-17 alpha-(6-mesyloxyhexynyl)estradiol was converted to the azide in 60-70% yield with NaN3/DMPU, then reduced to the corresponding amine (> 95% yield). Acylation with the N-hydroxysuccinimide esters of biotin, 5 carboxyfluorescein and 10-(3-sulfopropyl)-N-tosyl-N-(3- carboxypropyl)acridinium 9-carboxamide gave the title conjugates. The KDs of the tracers with an estradiol antibody ranged from 97-197 nM. PMID- 9871751 TI - Solid-phase synthesis of oligomeric deoxynucleic guanidine (DNG): a polycationic analogue of DNA. AB - DNG is a DNA analogue wherein the negatively charged phosphate backbone linkages have been replaced by achiral positively charged guanidinium linkages and has high affinity for complimentary DNA. The synthesis of these compounds in solution phase has been severely limited due to diminishing yields and solubility limitations. For the first time, an efficient solid-phase synthesis for oligomeric is reported. PMID- 9871752 TI - A 3D-QSAR study on ginkgolides and their analogues with comparative molecular field analysis. AB - Comparative molecular field analysis (CoMFA), a three-dimensional quantitative structure-activity relationship (3D-QSAR) paradigm was used to study the correlation between the physicochemical properties and the in vitro bioactivities of ginkgolide analogues. The correlation derived from CoMFA analysis has a good predictive capability. Based on the result of CoMFA analysis, we designed some compounds. Pharmacological assay indicated that three of these new designed compounds are 2 and 4 times more potent than that of ginkgolides. PMID- 9871753 TI - Synthesis of 5-thiomannose-containing oligomannoside mimics: binding abilities to concanavalin A. AB - 5-Thiomannose-containing oligomannoside mimics, 5SMan alpha(1,6)Man, 5SMan alpha(1,3)Man, 5SMan alpha(1,6)?Man alpha(1,3)Man?, Man alpha(1,6)?5SMan alpha(1,3)Man?, and 5SMan alpha(1,6)?5SMan alpha(1,3)Man?, were synthesized. Dissociation constants for the binding of these mimics to concanavalin A (ConA) were determined by a fluorescence anisotropy inhibition assay. Comparison of these data with those of the natural oligomannosides and with a crystal structure of the trimannoside-ConA complex established that replacing a ring oxygen atom with a sulfur atom causes about 1 kcal/mol decrease in the binding free energy when the ring oxygen is recognized with a hydrogen bonding. PMID- 9871754 TI - Synthesis of fluorescent substrates for protein tyrosine phosphatase assays. AB - Two fluorescent substrates for protein tyrosine phosphatase (PTPase) reaction were prepared by conjugation of commercially available O-phosphotyrosine and dansyl chlorides. They were hydrolyzed by CD45 tyrosine phosphatase, and proved to be useful for PTPase assay. PMID- 9871756 TI - Anti-Helicobacter pylori agents. 2. Structure activity relationships in a new series of 2-alkylguanidino-4-furylthiazoles. AB - SAR for antimicrobial activity against H. pylori was investigated in a new series of 2-alkylguanidino-4-furylthiazoles. Of the compounds obtained, cyclohexylmethyl and ethoxyethyl derivatives were identified as a novel class of anti-H. pylori agents which possessed potent and selective antimicrobial activity against H. pylori. These compounds also showed gastric antisecretory activity. PMID- 9871755 TI - Chemo-enzymatic synthesis of calcitonin derivatives containing N-linked oligosaccharides. AB - Eel calcitonin derivatives containing various N-linked oligosaccharides were chemo-enzymatically synthesized by the transglycosylation reaction of Mucor hiemalis endo-beta-N-acetylglucosaminidase (Endo-M) to a glycosylated calcitonin derivative [Asn(GlcNac)3]-CT in which N-acetyl-D-glycosamine (GlcNAc) is attached to the L-asparagine (Asn) residue of the peptide. PMID- 9871757 TI - Design and synthesis of isoxazole and isoxazoline linkage for replacement of nucleotide phosphodiester. AB - Synthetic routes for oligonucleotides which have isoxazoline or isoxazole linkage have been developed, and di- and tri-nucleotides with thymine bases have been prepared. PMID- 9871758 TI - Rapid synthesis of a 5'-fluorinated oligodeoxy-nucleotide: a model antisense probe for use in imaging with positron emission tomography (PET). AB - 5'-Deoxy-5'-fluoro-O4-methylthymidine was synthesized by the reaction of the corresponding 5'-O-tosylate with KF in the presence of Kryptofix [222] and coupled to a 5'-phosphoramidite-activated CPG-bound oligodeoxynucleotide. The sequence of reactions and purifications were accomplished within 4 h, a necessary condition of the development of radiofluorinated antisense oligodeoxynucleotide probe for use with PET. PMID- 9871759 TI - Synthesis and antibacterial activity of O-methyl derivatives of azalide antibiotics: II. 6-OME derivatives via clarithromycin. AB - Direct O-methylation of various derivatives of 9-deoxo-8a- and 9a-aza-8a-homo erythromycin (2',3'-bis-Cbz protected) gives 6-OMe derivatives only in a small number of special cases. The 6-OMe-azalides can, however, be synthesized beginning from clarithromycin. PMID- 9871760 TI - A new backbone of artificial enzymes obtained by cross-linkage of poly(ethylenimine). AB - Cross-linkage of the branches of poly(ethylenimine) (PEI) suppresses flexibility of the polymer as revealed by decreased affinity of the amino groups on PEI backbone towards proton or Ni(II) ion. The cross-linkage improves ability of the PEI derivative equipped with beta-cyclodextrin to deacylate an ester containing t butylphenyl moiety. PMID- 9871761 TI - Activation and stabilization of chymotrypsin in microdomains of poly(ethylenimine) derivatives. A model of in vivo environment. AB - Both activity and stability of alpha-Chymotrypsin (ChT) are substantially enhanced in the microdomains of laurylated or benzylated derivatives of poly(ethylenimine). EPR data revealed that the enhancement in activity of ChT is due to increase in the polarity of the microenvironment of Ser-195 caused by complexation of ChT to the polymer derivatives. PMID- 9871762 TI - Sulfonamide derivatives of benzylamine block cholesterol biosynthesis in HepG2 cells: a new type of potent squalene epoxidase inhibitors. AB - Sulfonamide derivatives of ene-yne benzylamine 1 have been prepared and identified as a new class of potent SE inhibitors having demonstrated activity in HepG2 cells as cholesterol biosynthesis inhibitors. PMID- 9871763 TI - High affinity, selective neurokinin 2 and neurokinin 3 receptor antagonists from a common structural template. AB - High affinity, selective hNK2 or hNK3 ligands can be prepared from the common template 1 in a few simple chemical operations. The hNK3 ligands 3 antagonise the calcium mobilisation caused by activation of hNK3 receptors expressed in CHO cells as measured using fura-2 microspectrofluorimetry. PMID- 9871764 TI - Synthesis and biological evaluation of 1,2-disubstituted carbonucleosides of 2 amino-6-substituted purine and 8-azapurine. AB - One, two-disubstituted carbocyclic nucleoside analogues bearing a 2-amino-6 substituted (chloro, hydroxy or amino) purine or 8-azapurine base were prepared by constructing the base about (+/-)-2-aminocyclopentane methanol, and their activities against a selection of viruses and tumor cells were determined in vitro. PMID- 9871765 TI - Search for alpha 1-adrenoceptor subtypes selective antagonists: design, synthesis and biological activity of cystazosin, an alpha 1D-adrenoceptor antagonist. AB - Two novel quinazolines (2 and 3) related to both prazosin and its open analogue 1 were synthesized, and their biological profile at alpha 1-adrenoceptor subtypes was assessed by functional assays in rat isolated tissues, namely prostatic vas deferens (alpha 1A), spleen (alpha 1B) and aorta (alpha 1D). Furthermore, the binding profile of 3 was assessed at native alpha 2 and D2 receptors, and cloned human 5-HT1A receptors, in comparison to prazosin, (+)-cyclazosin, 1 and BMY 7383. It turned out that the cystamine-bearing quinazoline 3 (cystazosin) has a reversed affinity profile relative to (+)-cyclazosin owing to a higher affinity for alpha 1D-adrenoceptors and a significantly lower affinity for the alpha 1A and alpha 1B subtypes. Furthermore, in comparison to BMY 7378, cystazosin (3) displays a much better specificity profile since it has lower affinity for D2 and 5-HT1A receptors. PMID- 9871766 TI - The synthesis of novel matrix metalloproteinase inhibitors employing the Ireland Claisen rearrangement. AB - Matrix metalloproteinase inhibitors of general formula (1) were synthesised by a route involving an Ireland-Claisen rearrangement which enables systematic modification of the substituent alpha to the hydroxamic acid. An analogue (12c) possessing an alpha-cyclopentyl group is a potent broad spectrum inhibitor that displays high and sustained blood levels following oral dosing in both the rat and marmoset ex-vivo bioassays. This compound and analogues are also potent inhibitors of TNF alpha release. PMID- 9871767 TI - Determination of 6s-trans conformation of retinal chromophore in sensory rhodopsin I and phoborhodopsin. AB - 8,16-Ethanoretinal 3 was synthesized from 2-methylcyclohexanone. From the binding experiments of 3 and 8,18-ethanoretinal 2 with apoproteins of sensory rhodopsin I and phoborhodopsin, it was found that retinal was incorporated as 6s-trans conformation in the both proteins. PMID- 9871768 TI - Asymmetric synthesis of Boc-N-methyl-p-benzoyl-phenylalanine. Preparation of a photoreactive antagonist of substance P. AB - The asymmetric synthesis of (S)-Boc-N-methyl-p-benzoyl-phenylalanine was performed by alkylation of sultam Boc-sarcosinate. The levorotatory sultam led to (S)-Boc-N-methyl amino acids with high optical purity. This photoreactive amino acid was incorporated into the sequence of a Substance P peptide antagonist. Comparison of the affinity and antagonistic properties of Biotinyl-apa-[D-Pro9, MePhe(pBz)10, Trp11]SP for human tachykinin NK-1 receptor demonstrated that this photoreactive antagonist should be a suitable tool for photolabelling studies. PMID- 9871769 TI - Synthesis and biological evaluation of new tetrahydronaphthalene derivatives as thromboxane receptor antagonists. AB - New polysubstituted tetrahydronaphthalene derivatives were prepared as thromboxane receptor (TP-receptor) antagonists. Within this series of compounds S 18886 has been identified as an orally active, highly potent antagonist with a very long duration of action in different species. PMID- 9871771 TI - The novel cyclopropapyrroloindole(CPI) bisalkylators bearing methoxycarbonyl and trifluoromethyl groups. AB - The novel 3-methoxycarbonyl-2-trifluoromethylcyclopropapyrroloindole (MCTFCPI) bisalkylators were synthesized and their antitumor activity was evaluated. Among these derivatives, 7f in which two MCTFCPI moieties are connected with a 5,5' bis(2-carbonyl-1H-indole) group, was found to exhibit more prominent cytotoxicity and antitumor activity than U-77,779 (bizelesin) (2). PMID- 9871770 TI - New tetrahydronaphthalene derivatives as combined thromboxane receptor antagonists and thromboxane synthase inhibitors. AB - A pyridine group was linked to the tetrahydronaphthalene moiety of the derivatives described in the preceding paper, to afford new combined thromboxane receptor (TP-receptor) antagonists and synthase inhibitors. The most interesting compound 2f inhibits TXA2 synthase with an IC50 value of 0.64 microM and the aggregation of human platelets with an IC50 value of 0.063 microM and shows a long duration of action in different species after oral administration. PMID- 9871772 TI - Novel D-ring analogues of podophyllotoxin as potent anti-cancer agents. AB - Several D-ring modified analogues of podophyllotoxin were prepared viz semi synthesis starting from naturally occurring podophyllotoxin and determined their in vitro anti-cancer activity. Most of the analogues have shown good activity towards human cancer cell lines. PMID- 9871773 TI - New synthetic inhibitors of microtubule depolymerization. AB - A new class of borneol esters that might be considered as biological analogs of paclitaxel regarding their action on microtubules has been found. By structure activity optimizations, compounds stabilizing microtubules much better than paclitaxel while showing a remarkably reduced cytotoxic activity were obtained. This dissoziation will open completely new therapeutic areas. PMID- 9871774 TI - The first conversion of camptothecin to (S)-mappicine by an efficient chemoenzymatic method. AB - Camptothecin has been converted for the first time to (S)-mappicine via mappicine ketone, which is the sole product of the microwave irradiation of camptothecin. Baker's yeast reduction of mappicine ketone yielded (S)-mappicine in high optical purity. PMID- 9871775 TI - Dimers of 5HT1 ligands preferentially bind to 5HT1B/1D receptor subtypes. AB - New dimers of known 5HT1 ligands (5HT, 1-NP or 8-OH-DPAT) have been prepared and evaluated at human cloned 5HT1B, 5HT1D and 5HT1A receptors. Binding experiments show that all these dimers have better affinities at 5HT1B/1D receptors than their corresponding monomeric ligands. Studies of inhibition of the forskolin stimulated c-AMP formation mediated by the human 5HT1B receptor show that hetero bivalent ligands [combining an agonist (5HT) with an antagonist (1-NP)] behave as partial agonists while the intrinsic activity of bivalent antagonists (combining two 1-NP residues) was found to be spacer dependent. Surprisingly enough, the dimer of 8-OH-DPAT 6 binds to 5HT1A, 5HT1B and 5HT1D receptors with similar high affinity. PMID- 9871776 TI - Synthesis and anticholinesterase activity of huperzine A analogues containing phenol and catechol replacements for the pyridone ring. AB - Based upon modeling results obtained using the crystal structure of huperzine A in complex with acetylcholinesterase (AChE), two novel analogues of this potent AChE inhibitor were designed with phenol or catechol rings replacing the pyridone ring. From the modeling studies, the catechol analogue appeared capable of replacing one of the crystallographic waters bridging huperzine with Tyr 130 and Glu 199 of AChE. The synthesis of these materials by use of a palladium catalyzed bicycloannulation strategy is detailed together with the results of AChE inhibition assays. PMID- 9871777 TI - Novel constrained CCK-B dipeptoid antagonists derived from pipecolic acid. AB - A new series of 4-substituted pipecolic acid derivatives was prepared and incorporated into dipeptoids. The resulting products behave as moderately potent CCK-B antagonists but their constrained structure and its comparison with structurally related compounds yield valuable information about the conformational requirements for optimal recognition of the CCK-B receptor by antagonists. PMID- 9871778 TI - Molecular lipophilicity potential by CLIP, a reliable tool for the description of the 3D distribution of lipophilicity: application to 3-phenyloxazolidin-2-one, a prototype series of reversible MAOA inhibitors. AB - The capacity factor of eleven derivatives belonging to a prototype series of 3 phenyloxazolidin-2-one, reversible MAO inhibitors, was measured and compared to the calculated log Pcalc using the CLIP package. We demonstrate that this Molecular Lipophilicity Potential (MLP) approach is a valuable tool to estimate log Pcalc of such compounds. PMID- 9871779 TI - Synthesis and biological activities of phenyl piperazine-based peptidomimetic growth hormone secretagogues. AB - A new class of potent, orally active phenyl piperazine-based GH secretagogues have been discovered from attempts to mimic the arrangement of the phenyl substituent in the spiroindanyl piperidine and spiroindoline sulfonamide privileged structures of 4 and 1, respectively. The best of these compounds, 18 (EC50 = 2.8 nM) is nearly as potent as MK-0677 for releasing GH from rat pituitary cells. PMID- 9871780 TI - Inhibition of human cytomegalovirus protease N(o) with monocyclic beta-lactams. AB - Monocyclic beta-lactams have been identified as potent and selective inhibitors of the human cytomegalovirus protease (HCMV) N(o). Two series of these inhibitors are described, a peptidyl series of compounds and non-peptidic molecules featuring lower molecular weights. The SAR work that lead to the discovery of these inhibitors, together with their synthesis is also disclosed. PMID- 9871782 TI - Attitudes of Kuwaiti parents toward physical punishment of children. AB - OBJECTIVE: The major aim was to describe parental attitudes to physical punishments and examine their sociodemographic correlates. A related aim was to assess the association of parents' own experience of physical punishment with attitudes to punishment of children. METHOD: A cross-sectional survey was conducted during the second week of December, 1996 in five general clinics covering the major administrative areas of Kuwait: 337 Kuwaiti mothers and fathers with at least one living child were contacted; 95% were successfully interviewed using a structured questionnaire. RESULTS: Eighty-six percent of parents agreed with physical punishment as a means of child disciplining. Agreement with punishment was higher in case of serious misbehaviors such as stealing (63%), sniffing glue and using drugs (77%). Multiple regression results showed that parent's lower level of education and Bedouin ethnicity were positively associated with agreement on physical punishment. Larger percentages of parents who had experienced physical punishments themselves agreed with such punishment to discipline their children, but this was not statistically significant. CONCLUSIONS: In recent years education has become widespread for both sexes. An inverse association between educational level and agreement on physical beating suggest that attitudes to this form of child disciplining are changing. Those with a Bedouin ethnic background still adhere more strictly to the traditional forms of child disciplining including physical beating. There is a need for conducting research on the possible negative psychosocial impacts of physical punishment in view of findings from other countries. PMID- 9871781 TI - Infants entering foster care compared to other infants using birth status indicators. AB - OBJECTIVE: Infants comprise nearly one fourth of all entries to foster care. Linkage of administrative birth and placement data can provide information about these infants that may otherwise be unavailable or difficult to obtain. METHOD: Statewide birth records and foster care placement histories were linked via probability matching. Logit regression was used to compare 26,460 maltreated infants who entered foster care between 1989 and 1994 with a random sample of 68,401 other infants born during that time frame. RESULTS: Infants in care were more than twice as likely to have single parents and be born with low birthweight, and twice as likely to have been born with a birth abnormality as other infants, controlling for other factors. The largest difference was in the eightfold increased likelihood for mothers of infants in care to have had no prenatal care. Infants in care were nearly three times as likely to be born into larger families (third or greater live births to the mother). Mothers of infants in care were more than twice as likely to be African American compared to White than mothers of other infants, while Hispanic and Other ethnic groups were underrepresented in the group of infants in care. Foreign born mothers, especially Hispanic women, were much less likely to have infants in care than they were to have children in the other group. CONCLUSIONS: Administrative datasets, while often limited in the number of variables they include and scope of their information, can be a valuable tool when used to understand demographics and frame questions for future research. Infants who enter foster care differ in substantial ways from other children. These findings have important implications for future research aimed toward targeting of child welfare services and supports. PMID- 9871783 TI - Family of origin history, psychological distress, quality of childhood memory, and content of first and recovered childhood memories. AB - OBJECTIVE: Individual differences in quality of childhood memory and recovered memories from childhood remain poorly understood. Therefore, this study tested several hypotheses which may help account for the large amount of variation that individuals report in the general quality of their childhood memory and the valence of the memories that many individuals report recovering from their childhoods. It was hypothesized that family of origin dysfunction would be associated with poorer childhood memory, that current depressed mood would be associated with impaired childhood recall and the recall of negative memories, and that the content of recovered childhood memories would be disproportionately negative because they include a significant number of memories which had been repressed or dissociated. METHOD: Questionnaires were administered to 553 college students, 27% of whom reported a history of child abuse. RESULTS: The participants reported substantial variation in the general quality of their childhood memories and also a wide variety of different types of experiences for both their first childhood memories and the recovered memories that most of them had from their childhoods. Weak associations were found between family of origin dysfunction and poorer general quality of childhood memory, but the study as a whole resulted in few significant findings. CONCLUSIONS: Only weak support was found for some of the factors that have been hypothesized to distort autobiographical memory. The substantial individual variation in childhood memory that has been reported by college student samples remains poorly understood. PMID- 9871784 TI - Memories of childhood sexual abuse: a survey of young adults. AB - OBJECTIVE: To explore the prevalence of, characteristics of, and factors associated with forgetting of childhood sexual abuse memories in a large non clinical sample (N = 1712). METHOD: Using an anonymous survey, we asked respondents about (a) the nature and severity of their childhood abuse; (b) the continuity of their abuse memories; and (c) their experiences with others suggesting to them that they might have been abused. RESULTS: A substantial minority of victims in our sample reported having temporarily forgotten their childhood sexual abuse. Forgetting was largely unassociated with victim or abuse characteristics. Compared to individuals who always remembered their abuse, however, individuals who temporarily forgot were more likely to report that someone had suggested to them that they might have experienced abuse. Those who received such suggestions were particularly likely to suspect that they may have experienced childhood sexual abuse that they do not yet remember. CONCLUSION: Forgetting may be less common than implied by earlier estimates from clinical samples, yet it is not uncommon. Also, a sizable minority of the population is wondering whether they have experienced unremembered abuse, and these suspicions are linked to having encountered suggestions from others. We discuss the implications of these findings for understanding the phenomenon sometimes labeled repression. PMID- 9871785 TI - Child sexual abuse: victim age, victim gender, and observer gender as factors contributing to attributions of responsibility. AB - OBJECTIVE: The purpose of this study was to investigate the effects of victim gender, and observer gender on the tendency to attribute responsibility for extrafamilial child sexual abuse to the victim and the nonoffending parents. METHOD: A 2 (Victim Age) x 2 (Victim Gender) x 2 (Observer Gender) between subjects design was employed. Undergraduate students (N = 145) read a vignette describing a sexually abusive interaction between an adult male neighbor and a child. In this vignette, the child's gender and age (6 years old, 13 years old) varied. After reading the vignette, participants used a 5-point scale to indicate the degree to which they believed the victim and the parents (a) were responsible for, (b) were to blame for, (c) caused, and (d) could have prevented the abuse. RESULTS: Greater responsibility was assigned to older than younger victims. Both parents were ascribed similar levels of responsibility, and were ascribed greater responsibility when the child victim was younger than older. Male observers attributed greater responsibility and causality to the victim and the parents than did female observers. CONCLUSIONS: The results indicate that responsibility attributions directed toward the victim and the nonoffending parents may be a function of the victim's age. In addition, the findings support previous research suggesting that male observers may tend to hold victims more responsible for their abuse than female observers. Implications for treatment and research are discussed. PMID- 9871786 TI - Examining the validity and reliability of childhood abuse scales: putting The Courage To Heal to the test. AB - OBJECTIVE: The main purpose of the current study was to examine the validity and reliability of The Courage To Heal Workbook checklist (Davis, 1990) in part, through examining the internal consistency and studying whether the CTHC distinguished between the participants reporting sexual abuse histories and those who did not. METHOD: Two hundred and seventy-nine college students were surveyed utilizing the symptom checklist from the Courage to Heal Workbook, Trauma Symptom Checklist-40 (TSC-40), Abusive Behavior Inventory (ABI), and questions related to participants' beliefs that they had been abused as a child. RESULTS: Results indicated that the Courage to Heal Checklist (CTHC) has robust reliability (alpha = .97) and can significantly discriminate between reported abuse survivors and nonsurvivors. Fifty-five percent of those participants reporting sexual abuse, 47% of those reporting physical abuse, and 34% of those reporting emotional abuse indicated a period of time during which they did not remember the abuse. CONCLUSION: Results indicate that the CTHC reliably differentiated between participants reporting past sexual abuse and those who did not. Mounting evidence continues to support that adult symptom profiles are able to indicate that some form of trauma is likely to have occurred in the past; however, differentiating between types of abuse based on symptom profiles may be impossible. PMID- 9871787 TI - Decisions and attitudes concerning child sexual abuse: does the gender of the perpetrator make a difference to child protection professionals? AB - OBJECTIVE: An investigation was conducted into whether child protection investigators, specifically social workers and the police, are as likely to take seriously a case of child sexual abuse if it had been perpetrated by a female rather than a male. Also, to examine whether the decisions relating to female perpetrated abuse were predicted by participants' sex role perceptions of women and their attitudes concerning women's sexualized behavior towards children. METHOD: Participants advocated decisions in response to four hypothetical case of child sexual abuse in which the perpetrator was either male or female. The female perpetrators were then rated on femininity and masculinity characteristics and attitudes concerning women's sexualized behavior toward children were assessed. RESULTS: Following male--rather than female--perpetrated sexual abuse, case registration and imprisonment of the perpetrator was considered more appropriate by all participant groups; male social workers also considered social services involvement and investigation as more appropriate. A substantial number of decisions concerning female perpetrated abuse were predicted by participants' attitudes. CONCLUSION: While child protection professionals considered child sexual abuse perpetrated by females to be a serious issue warranting intervention, a number of advocated decisions suggested that they did not consider female-perpetrated abuse to be as serious as male-perpetrated abuse. The implication is that victims of sexual abuse perpetrated by a woman may be less likely to receive the protection afforded victims of male-perpetrated abuse. Furthermore, professionals' practices may be inadvertently perpetuating the view that female child sexual abuse is rare or less harmful than abuse carried out by males. PMID- 9871788 TI - Child extrafamilial sexual abuse: predicting parent and child functioning. AB - OBJECTIVE: This study evaluated the emotional and behavioral adjustment of parents and children within 3 months and 1 year after the discovery of child extrafamilial sexual abuse. METHOD: Ninety-two case parents (63 mothers, 29 fathers) and 56 children were compared to a nonclinical comparison group of 136 parents (74 mothers, 62 fathers) and 75 children. Parent adjustment was assessed using self-report measures while child functioning was assessed using a combination of child-, parent- and teacher-report measures. RESULTS: Mothers, fathers and sexually abused children experienced clinically significant effects both initially and at 12 months post-disclosure. Children's perceptions of self blame and guilt for the abuse and the extent of traumatization predicted their self-reported symtomatology at 3 months and 1 year post-disclosure. Child age and gender also significantly contributed to the prediction of many of the child outcome measures. No abuse-related variable was related to any child self-report measure. Mothers' satisfaction in the parenting role, perceived support and intrusive symptoms predicted their initial emotional functioning. Avoidant symptoms, child's internalizing behavior and mothers' initial emotional functioning were significant predictors of longer-term emotional functioning. CONCLUSIONS: Results emphasize the need to address children's abuse-related attributions and underscore the need to expand our focus beyond the child victims to the traumatized families. PMID- 9871789 TI - Incest survivor mothers: protecting the next generation. PMID- 9871790 TI - [In commemoration of the 150th anniversary of the foundation of the Societe de Biologie]. PMID- 9871791 TI - [Foundation and brief chronology of the Societe franco-japonaise de Biologie]. PMID- 9871792 TI - [The Socieete franco-japonaise de Biologie since the war (creation of decentralized meetings)]. PMID- 9871793 TI - [One hundred and fifty years of the Societe de Biology]. PMID- 9871794 TI - [Genetics and politics]. PMID- 9871795 TI - [From the glycogenic function of the liver to gene regulation by glucose]. AB - Glucose, that Claude Bernard has demonstrated in 1850 to be synthesized and secreted by the liver, is an important regulator of gene transcription in all types of organisms. In vertebrates, it especially regulates transcription of metabolic genes in the liver and fat tissue, activating genes encoding enzymes and regulators of the glycolytic and lipogenic pathways. Working with the L-type pyruvate kinase gene we have found that in hepatocytes glucose-dependent gene regulation requires: Presence of the GLUT2 glucose transporter, necessary to allow for an effective depletion in glucose 6-phosphate (G-6P) under gluconeogenic conditions. Phosphorylation of glucose to G-6P assured either by insulin-dependent glucokinase or by another hexokinase isoform. Most likely, entry of G-6P in the pentose phosphate pathway. Modulation of a kinase/phosphatase cascade, in particular inhibition of the 5'AMP-activated protein kinase. Signalling through a glucose response complex assembled onto a glucose-response element (GIRE) located in regulatory regions of glucose responsive genes. The activators USF belong to the complex, and are required for a normal gene activation by glucose, as evidenced from the phenotype of knock-out mice deficient in USF. The study of USF-defective knock-out mice suggest that USF could be involved in nutritional activation of a whole class of genes regulated by glucose, and not by insulin itself. In particular, lipogenic genes and the ob gene, encoding the leptin satiety hormone, are abnormally responsive to diet in USF-/- mice. The transactivation potential of USF would be modulated by a glucose sensor system implying the COUP-TFII transcription inhibitor. The main role of insulin in the glucose response of genes like the L-PK gene is to induce the glucokinase gene. Glucagon, through cyclic AMP, inhibits L-PK gene transcription mainly through activation of PKA. The PKA catalytic subunit could act by phosphorylating member(s) of the glucose-response complex, or of contiguous transcription factor, e.g. HNF4. In conclusion, through a pluridisciplinary approach ranging from Claude Bernard-derived biology to modern molecular biology, important progress have been made during the last years on the mechanisms of the regulation of gene transcription by glucose in vertebrates. PMID- 9871796 TI - [From Claude Bernard to the regulatory system between the hypothalamus and the periphery: implications for homeostasis of body weight and obesity]. AB - The concept of interrelationships between the central nervous system and the periphery aimed at maintaining normal body weight homeostasis has been strengthened by the discovery of hypothalamic neuropeptide Y (NPY) and adipose tissue leptin. NPY, when infused intracerebroventricularly in normal animals produces hyperphagia and hormono-metabolic changes (hyperinsulinemia, hypercorticism) channeling nutrients preferentially toward lipogenesis and storage in adipose tissue and away from their utilization by muscles (muscle insulin resistance). Storage in NPY-infused rats is further favored by the observed decrease in the expression of uncoupling proteins. NPY-induced hyperinsulinemia and hypercorticosteronemia also promote leptin over-secretion. Released leptin, acting within the hypothalamus, decreases hypothalamic NPY levels (probably those of other hypothalamic neuropeptides as well), food intake, insulinemia, insulin sensitivity of white adipose tissue, while increasing that of muscles. Leptin acting centrally additionally favors the expression of uncoupling protein 1, 2, and 3, in keeping with an eflect on energy dissipating mechanisms. The respective hormono-metabolic eflects of NPY and leptin maintain a normal body homeostasis. In most obesity syndromes, the functional relationships between NPY and leptin are altered. Due to hypothalamic leptin receptor mutations or dysfunctions, leptin cannot exert its eflects: NPY levels (possibly those of other neuropeptides) remain elevated, maintaining excess storage, insulin as well as leptin resistance. PMID- 9871797 TI - [Renewal and stability]. PMID- 9871798 TI - [Signal transduction and thyroid pathology: environment and genetics]. PMID- 9871799 TI - [The challenge of science at the outset of the third millenium]. AB - Compared to others species Homo sapiens is physically underprivileged as much by his relative weakness than by his lack of natural defense. Only his mental abilities allowed him to cope with the various dangers which threaten him and to rule over the biosphere that he managed to modified for his own benefit. The neolithic revolution which saw the generalization of agriculture and the breeding of animals offered a considerable quantities of resources which lead to a strong demographic growth. All this produced a successions of discoveries which improved his quality of life. Today after an era of quantitative growth the production of the agroalimentary sector enter a qualitative period in which the aim is note only to increase the production of the traditional resources but, by the alteration of the genetic heritage, to improve their properties. PMID- 9871800 TI - [Ethical problems connected with developmental biology]. PMID- 9871801 TI - [New procreations in the origin of a new biology]. PMID- 9871802 TI - [Reflections on the origin and evolution of life]. PMID- 9871803 TI - [Role of science in the formulation of social policy on drugs]. AB - Psychoactive drugs are used almost universally for the pleasure and benefits which they can provide, but they also cause sufficient harm that most societies have adopted policies to control and limit the amount of use. Science is increasingly called upon to provide a rational basis for these policies. However, the biological sciences and modern sociology have fundamentally different approaches to such issues, the former being based on the concept that there is an external reality which can be discovered only by objective means, the latter holding that social problems are defined strictly subjectively and can not be separated from the values and ideologies of the researchers. Since social policy affects all members of a society, and must reflect all of their attitudes, values and traditions, science can contribute only facts and probabilities, but society as a whole must assign the values and make the required choices. PMID- 9871804 TI - [Chemokines and the regulation of hematopoiesis]. AB - Chemokines are a large family of cytokines that act not only as immune and inflammatory regulators but also as regulators of hematopoiesis. Two major subfamilies of chemokines are distinguished on the basis of whether the first two cysteines are separated by a single residue (CXC) or three residues (CX3C) or they are adjacent (CC) or there is a single C. The Macrophage Inflammatory Protein 1 alpha (MIP-1 alpha), which belongs to CC family is a powerful inhibitor of hematopoisis in vitro and in vivo. The sub-family CXC comprises two main groups. The first sub-group includes the ELR chemokines, in which interleukin-8 (IL-8) is the most prototypic and possesses suppressive activities on hematopoiesis. Platelet Factor 4 (PF4) belongs to the sub-group of non-ELR CXC chemokines. PF4 acts as an inhibitor of hematopoiesis, particularly of the megakaryocytopoiesis. Recently, it has been shown that a peptide of PF4, 34-58 which does not contain the site of heparin binding, is able to inhibit the growth of hematopoietic progenitors in vitro, providing evidence for a model of heparin dependent and independent pathways of PF4 action on hematopoiesis. PF4 can reduce the chimiosensitivity of hematopoietic cells in mice treated by the cytotoxic drug 5-Fluorouracyl, suggesting a potential clinical application of PF4 in cancer therapy. PMID- 9871805 TI - [The human genome project initiative in China]. PMID- 9871806 TI - [Glycolytic enzyme defects and neurodegeneration]. AB - This study was devoted to the continued search for an explanation of the neurodegeneration found in a severely TPI deficient Hungarian patient whose brother with genomically completely identical TPI defect was completely free of neurological disorders. The changes found in the molecular species composition of the major PL subclasses and the decrease in PE plasmalogens explain the earlier round increase in membrane fluidity interfering thereby with the physiological function of membrane enzymes, receptors, signal transduction, protein-protein interactions and vesicle fusion. Plasmalogens have also the capacity to protect against oxidative stress, that is deemed to contribute to neurodegenerative processes. The presence of chronic oxidative stress was well reflected in the decreased levels of GSH and alpha-tocopherol in the affected brothers. Decrease in plasmalogens have been described recently in Zellweger's syndrome, in other peroxisomal neurodegenerative disorders, in demyelinating processes and in Alzheimer's disease. The brain in normal individuals is highly enriched in plasmalogens. The pathological decrease found in TPI deficient lymphocytes will presumably be more pronounced in excitatory tissues. The recently described role of expanding nucleotide triplets in the development of neurodegeneration is suggested to result through the selective binding via their polyglutamine repeats to GAPDH. The role of GAPDH in TPI deficiency may be of crucial help in the elucidation of the development of neurodegeneration, since the enzymatic defect of TPI can be partially bypassed by means of the HMP shunt which generates GAP via GAPDH without the participation of TPI. Considering the results found in TPI deficiency in comparison to the new literary findings in different neurodegenerative diseases the following pathomechanism may be proposed. The protein products of the defective genes due to their abnormal steric structure bind GAPDH in a different manner or in differing quantity than their normal counterparts. The PL composition and the resulting differences in the biophysical properties of the cell membranes have crucial impact on these protein-protein interactions and on the activity of enzymes and membrane transport functions. The plasmalogen decrease impairs the protection against oxidative stress with consecutive worsening of the neurodegenerative process. The final common pathway to neuronal death leads through destabilization of intracellular Ca2+ homeostasis via elevation of intracellular Ca2+ to apoptosis. The most important conclusion is that lipids are not an inert environment of membrane proteins. Unravelling of the pathogenesis of neurodegeneration needs more concerted investigation of the interactions between genetic changes with biophysical and biochemical cell membrane lipid alterations. PMID- 9871807 TI - [Factors affecting the trans-endothelial accumulation of atherogenic plasma proteins in artery walls]. AB - Our experiments were done to find out whether there are factors which influence the passage of the two major "risk factor proteins" LDL and fibrinogen, other than their plasma concentrations, from the blood into the arterial walls, where their accumulation is associated with atherogenesis. The results suggest that administration of a remarkable variety of pressor agents over a few days accelerate the uptake of both proteins by arterial walls, and that, in contrast, the process is no faster in rats that have been spontaneously, i.e., genetically, hypertensive for about 3 months. Considering our experimental findings in relation to human atherosclerotic disease, it is interesting that the risk of coronary heart disease and stroke is increased more than additively when both LDL or fibrinogen levels and systolic or diastolic blood pressures are high (18,46). If our work should point to some mechanistic connection between blood pressure and the accumulation of atherogenic plasma proteins in arterial walls, it would provide, at least in principle, an explanation for the epidemiological facts. PMID- 9871808 TI - [Recent developments of genetic markers in legal practice]. AB - The human identification in forensic science is currently based upon the study of highly polymorphic systems with mendelian transmission: the STRs (Short Tandem Repeats). These genetic markers are three to seven bases repetitive sequences spread all over the genome. They are detected by using the Polymerase Chain Reaction (PCR). This analysis method makes it possible to amplify several different loci from very low quantities of genomic DNA (Desoxyribonucleic Acid) in very short periods of time without using any radioactive substances. Interpretation of the results is simple because the amplified fragments are of a known size. The alleles known in each system with discrete values allow the calibration of the revealing gel and the populations studies; moreover, they may be entered into a database. The fragility of this test implies specific precautions with the Laboratory's organization in order to avoid any kind of contamination. Both the establishment of a quality assurance responding to the ISO 9002 standard together with internal quality controls would ensure that these tests are reliable and repeatable in the opinion of the Courts. Only authorized laboratories may perform DNA testing on the occasion of legal proceedings. PMID- 9871810 TI - [Role of cytokines in the pathogenesis of gastrointestinal diseases associated with Helicobacter pylori infection]. AB - It is well known that Helicobacter pylori can cause gastritis, gastroduodenal ulcers and malignant diseases. The infiltration of polymorphonuclear leukocytes is recognized in the lesions of these diseases, and the infiltration disappears by antibiotic therapy. However, it is not yet clarified how Helicobacter pylori induces the formation of lesions including leukocyte infiltration. Recently, we have confirmed that several kinds of cytokines are expressed in the gastric biopsy specimens of gastroduodenal diseases. Especially, it is conjectured that chemokines such as interleukin-8 (IL-8) which are expressed in the specimens, induce leukocyte infiltration, gastric mucosal inflammation and gastroduodenal ulcers. It is possible that Helicobacter pylori CagA gene is closely related with IL-8 expression because this cytokine is more strongly expressed in the specimens from the patients infected with CagA-positive Helicobacter than those with CagA negative one. PMID- 9871809 TI - [Biosynthesis and transport of sterols in the yeast Saccharomyces cerevisiae]. AB - The yeast Saccharomyces cerevisiae was a powerful tool in the identification of the structural genes involved in sterol biosynthesis in eucaryotes. Among 20 genes, 16 were isolated by genetic techniques using either complementation of mutants or overexpression strategy using specific inhibitors. In spite of this good knowledge concerning the genes of the pathway, little is known about the regulation of the isoprenoid/steroid biosynthetic pathway. However, the existence of two genes encoding HMG-CoA reductase in yeast genome suggests strongly that this enzyme could play a fundamental function in regulation, such as in plants and mammals. The regulation mechanisms could also involve sterol trafficking and storage. Indeed, one enzyme in the pathway, the sterol-C24-methyl transferase is localized in lipid particles that correspond to the storage form of steryl esters. Yeast cells are impermeable towards exogenous sterols in aerobiosis and become permeable in anaerobiosis when ergosterol synthesis is precluded by the absence of molecular oxygen. This phenomenon called aerobic sterol exclusion is dependent on the hem status of the cell. One gene, named SUT1 was identified that directs aerobic sterol uptake in yeast SUT1 gene and his partner SUT2 present strong features common to yeast transcription factors and could regulate the expression of genes involved in sterol uptake or intracellular trafficking. PMID- 9871811 TI - [Osmotic fragility test of erythrocytes with a coil planet centrifuge]. AB - The measurement of osmotic fragility of erythrocytes has been applied to the diagnosis of hemolytic diseases, studies of membrane permeability and alternations leading to destruction of erythrocytes. Almost 30 years have gone by since the coil planet centrifuge system was devised for measuring the osmotic fragility of erythrocytes. Many excellent studies by means of this centrifuge system have been published. Prominent investigations are reviewed as follows: in relation to the osmotic fragility, various liver diseases, angina pectoris, trapping the aged erythrocytes, tumor, lactic acid, unsaturated fatty acids, free cholesterol, exercise and lead exposure level were examined. PMID- 9871812 TI - [Increase of circulating CD3+CD4-CD8-CD19+ cells in the latent phase of HIV-1 infection]. AB - Having reported that HIV-1-infected T cell lines are rescued as CD4- from cytolysis by human complement factor B, we now show the presence of an in vivo counterpart of such CD4- T cells by demonstrating the circulating CD3+ CD4- CD8- CD29+ cells in the blood of seropositive subjects (n = 91, classified by the immunologic scale scores 0, 1, 2 and 3). The cell population was found to be significantly increased in the early phase of infection in score 0: 195/mm3 (p < 0.005) and in score 1:376/mm3 (p = 0.001). With the infection progressing to score 2, the cells decreased to 220/mm3 (p < 0.001) and finally to the same range: 101/mm3, as that of uninfected subjects. Further elucidation of the mechanism of the appearance and disappearance of that population in vivo could help to elucidate protective immunologic processes. PMID- 9871813 TI - [Induction of epidermal growth factor (EGF) in the urogenital sinus by a brief treatment with androgens]. AB - We have investigated the induction of epidermal growth factor (EGF) in the urogenital sinus of fetal mice when the sinuses were cultured with testosterone. Immunofluorescence study revealed that almost no EGF is detectable in the pre incubated sinuses, whereas EGF is induced in the sinus mesenchyme as early as 2 hours after the testosterone treatment. The results support the hypothesis that paracrine-like factors secreted from the sinus mesenchyme activated by androgens may be EGF or EGF-like growth factors that stimulate the sinus epithelium to induce prostatic buds. PMID- 9871814 TI - Anxiety as a predictor of response to interpersonal psychotherapy for recurrent major depression: an exploratory investigation. AB - Major depression and anxiety frequently co-occur, but the implications for psychological treatments have rarely been studied. We examined predictors of acute response to interpersonal psychotherapy in 134 consecutively treated female outpatients with recurrent unipolar depression. Women who failed to remit with interpersonal psychotherapy alone experienced higher levels of somatic anxiety, were more likely to meet criteria for lifetime panic disorder, were more likely to meet criteria for nonendogenous or nonmelancholic depression, and reported greater vocational impairment, higher levels of global severity, a longer duration of the index episode, and, somewhat surprisingly, lower levels of social impairment at pretreatment evaluation. A series of backwards stepping logistic regression analyses showed higher levels of baseline somatic anxiety and social functioning to be the most consistent predictors of nonresponse. Our findings strengthen existing evidence that concomitant anxiety can adversely affect the outcome of interpersonal therapy for depression. PMID- 9871815 TI - Six-month outcomes for MRI-related vascular depression. AB - The purpose of this paper is to estimate the relative probabilities of 6-month recovery from an index episode of major depression for subjects with and without MRI-confirmed vascular brain changes. In this cohort study, 57 depressed subjects from the Duke University Mental Health Clinical Research Center (MHCRC) for Depression in Late Life who presented with MRI-related vascular and non-vascular depression were followed for 6 months, and the rates of recovery in the two risk groups were compared. Overall, the recovery rate in this sample was 57.9%. Subjects with MRI-related vascular depression demonstrated outcomes similar to subjects with non-vascular depression (crude RR = 0.67 [0.32, 1.43]). There was a trend that demonstrated that MRI-related vascular depression placed elderly subjects and subjects with first onset of depression after age 40 at increased risk of non-recovery. The study demonstrates overall no significant difference in course between patients with and without vascular depression. It also suggests that patients with vascular depression may have a different course depending on their age and age of onset of the disease. PMID- 9871816 TI - Comparison of the tolerability and efficacy of citalopram and amitriptyline in elderly depressed patients treated in general practice. AB - The enhanced sensitivity of the elderly to the side effects produced by tricyclic antidepressants (TCAs), and the frequency and type of adverse events, have made the treatment of depression in this group difficult. The selective serotonin reuptake inhibitors (SSRIs) have been reported to produce significantly fewer undesirable side effects and display better tolerance than TCAs. We compared the therapeutic actions and side effects produced by citalopram, the most selective SSRI available, with amitriptyline in a group of elderly patients (aged 65 and older) diagnosed with major depression. In a double-blind, double-dummy, parallel group, multicenter comparison of citalopram (20 or 40 mg/day) and amitriptyline (50 or 100 mg/day), patients who did not respond to placebo during a 1-week single-blind phase were randomly assigned to receive citalopram or amitriptyline for 8 weeks. Efficacy measures included the Montgomery-Asberg Depression Rating Scale (MADRS), the Hamilton Depression Scale (HAMD), and Clinical Global Impressions. Both drug treatments produced equivalent time-related declines in severity of depression, so that by 8 weeks slightly more than 50% of the patients in each group experienced marked recovery, defined as MADRS scores < or = 12. Amitriptyline produced a greater overall incidence of adverse events, including a significantly higher (P < 0.001) percentage of patients reporting dry mouth (34% vs. 7%), as well as a significantly higher (P < 0.02) incidence of somnolence. Constipation and fatigue also occurred more frequently in the amitriptyline than in the citalopram group. For only one event (nausea) did the citalopram group report a significantly greater (P = 0.012) incidence (12.8% vs. 4.8%). On the basis of these results, it was concluded that citalopram is as effective an antidepressant as amitriptyline in the treatment of the depressed elderly. Because of its low incidence and low magnitude of side effects, citalopram seems especially useful in private practice. PMID- 9871817 TI - Relationship between interpersonal psychotherapy problem areas with temperament and character: a pilot study. AB - OBJECTIVE: To examine the relationships between dimensions of personality assessed by using the Temperament and Character Inventory and presenting problem areas in Interpersonal Psychotherapy. METHOD: One hundred two depressed patients completed the Temperament and Character Inventory (TCI), and Interpersonal Psychotherapy (IPT) problem areas were assessed using a clinician-rated questionnaire. RESULTS: Following multiple regression analysis, patients rated as having problems with grief scored low on self transcendence, those with disputes had low cooperativeness, those with role transitions had low novelty seeking, and those with deficits had low reward dependence. CONCLUSIONS: Both temperament and character may be relevant to the nature of presenting problems related to Interpersonal Psychotherapy. An understanding of how personality is relevant to presenting interpersonal problems may provide both a framework and a focus with which to work in this form of psychotherapy. PMID- 9871818 TI - Familial aggregation of delusional depression: re-examination in a recent family study. AB - BACKGROUND: Delusional (D-MDD) and nondelusional depression (ND-MDD) differ in clinical presentation, biological abnormalities, course of illness, and treatment response. Family data, however, have been less consistent regarding differential risk both for any major depression (MDD) and specifically D-MDD in relatives of D MDD probands. In an earlier family study, we observed a 1.5-fold increase in rates of any MDD, specificity of transmission of D-MDD, and increased rates of bipolar disorders in relatives of D-MDD compared to relatives of ND-MDD probands. In a new family study, we attempted to replicate these findings. METHOD: A family study of 361 directly interviewed adult first-degree relatives (FDRs) of 163 probands (118 with MDD and 45 screened normal controls) was used to examine familial aggregation of any MDD, D-MDD, and bipolarity by proband delusional status. RESULTS: Compared to FDRs of ND-MDD probands, FDRs of D-MDD probands were at modestly increased risk for any MDD. These results were unaffected by adjustment for proband ascertainment source, comorbidity, or whether probands had chronologically primary MDD. There was a trend toward increased rates of broadly defined bipolarity (bipolar I, bipolar II, or cyclothymia) in FDRs of D-MDD compared to FDRs of ND-MDD probands. CONCLUSION: Results from the present study were broadly consistent with those from our previous work. While other lines of evidence for D-MDD as a distinct subtype are more compelling than family data, it would be of methodologic interest to identify sources of inconsistency across studies in findings concerning the familial aggregation of delusional depression. PMID- 9871819 TI - Validation of the hypomania interview guide-seasonal affective disorder (HIGH SAD) version in patients with rapid cycling bipolar disorder. AB - We validated the Hypomania Interview Guide-Seasonal Affective Disorder version (HIGH-SAD) in patients with rapid cycling bipolar disorder (RCBD). Fourteen outpatients were rated on six separate occasions (total = 84 visits). On each visit the patients were rated with the HIGH-SAD and the Young Mania Rating Scale (YMRS) in a counterbalanced order. Clinical assessment was completed at the end of the visit by the treating psychiatrist. Patients were assessed as hypomanic/manic on 22 of the visits. Pearson correlation coefficient between the YMRS total scores and the HIGH-SAD total scores for those 22 visits in which patients were hypomanic/manic was r = 0.629 (P < 0.05) and for all visits was r = 0.769 (P < 0.0001). Analysis with only one rating per patient yielded a Pearson correlation coefficient of r = 0.792 (P < 0.0004). We found that the HIGH-SAD was a valid scale for the measurement of hypomania in patients with RCBD. However, the scale does not differentiate hypomania from mania in this group of patients. PMID- 9871820 TI - Patient assessment: validation of a nursing instrument. AB - The aim of this study is to evaluate the reliability and validity of the Byron Physical Assessment Framework (BPAF). The BPAF is a systems based checklist of physiological measurements, signs, and symptoms designed to structure and document the assessment of a patients physical condition by nursing staff. Initially the BPAF was refined using extensive literature review and expert opinion to improve the comprehensiveness and clarity for its intended purpose. As a result the content validity of the BPAF was supported. Inter-rater reliability between both expert-expert and novice-expert pairings was assessed. A total of 68 assessments were conducted by pairs of qualified nurses with patients on general medical and surgical wards and a nursing-led unit in one London Hospital. Reliability of the dichotomous data items of the BPAF was found to be generally good (kappa > 0.6) with only one item showing a poor reliability (kappa < 0.20). Reliability for continuous items such as pulse and respiratory rate was surprisingly low with evidence of large variation between raters in addition to systematic bias. With relatively little teaching, novice assessors were able to use the BPAF and achieve good inter-rater reliability with expert assessors although this was lower than the reliability of the expert diads. The utilisation of the BPAF to fulfil intended purpose was assessed by examining completed assessments and the outcome in terms of nurses' actions in light of new abnormal findings. Both were found to be fair, showing that the BPAF does affect the actions of nurses although it could be utilised more. PMID- 9871821 TI - Telephone or face-to-face interviews?: a decision made on the basis of a pilot study. AB - De Vaus (1991) highlights five main considerations that may be key factors in decisions about whether to use telephone or face-to-face interviews for survey work: response rates, ability to produce representative samples, effects on interview schedule design, quality of responses and implementation problems. De Vaus' discussion of these five issues is outlined at the start of this article. The five issues are then applied to the experiences of researchers conducting a study on continence care. Description and discussion of pilot interviews, which explored both interview modes, are followed by a similar examination of the main study which employed telephone interviews. Ideas in this discussion are supported and challenged by reference to other publications on the subject of telephone and face-to-face interviews. The success of the decision to use telephone interviews in the main study is evaluated and recommendations are made. PMID- 9871822 TI - One step towards the understanding of hope: a concept analysis. AB - Hope is an essential concept in nursing which has not been satisfactorily explored. The purpose of this article is to elucidate the concept of hope, using a technique of concept analysis described by Walker and Avant (1995). The critical attributes revealed were; future-orientation, positive expectation, intentionality, activity, realism, goal-setting, and inter-connectedness. Various illustrative cases are presented as well as antecedents and outcomes. The discussion highlights some critical aspects of the method used and the results of this analysis may serve as a guide for further empirical research. In order to grasp the true nature of hope, complementary research studies employing a variety of methodological approaches are needed. PMID- 9871823 TI - The behaviors of AIDS patients toward their nurses. AB - The purpose of this study is to identify the behavioral responses of hospitalized patients with HIV/AIDS to nursing care providers. The critical incident technique, developed by Flanagan (1954) was used to obtain a listing of the behavioral responses. Patients were asked to recall brief descriptions of caregiving events. A purposive sample included 118 men and women with HIV/AIDS from broad socioeconomic and cultural backgrounds. A total of 273 critical incidents yielded a listing of 393 behaviors. The analysis of data was facilitated by a computer program which allowed for the creation of coding systems and refinement of coded items into behavioral response categories. The inductive content analysis yielded 10 major response categories: participate, anger, appreciate, come close, stay away, match respect, match disrespect, dependent, complaint, and self care. In the largest category, 41% of the patients described ways in which they participate actively in their nursing care. These behavioral responses sharply contrast with current literature which continues to place a negative emphasis on the attitudes and behaviors of nurses. One third of the patients listed angry behaviors which were directed at nurses. Behavioral descriptions of anger reflected increased irritability with advancing illness, intense psychological responses toward an AIDS diagnosis, or a violent and angry style of relating to others in street settings. Two of the response categories describe the reciprocal nature of nurse-patient interactions. By becoming aware of patient responses, nurses will obtain a greater understanding of what changes would influence outcomes in patient behavior. PMID- 9871824 TI - Assessing occupational stress in psychiatric nurses using the full job strain model: the value of social support to nurses. AB - The present research examined the effects of occupational stress in psychiatric nursing on employee well-being using the full Job Strain Model. The Job Strain Model was assessed for its ability to predict employee well-being in terms of job satisfaction and mental health. The original Job Strain Model was expanded to include social support based on previous research concerning the impact of social support on well-being. In the present study, both work support and non-work were assessed for their contribution to well-being. The results of this study indicate that the full Job Strain Model can be used to significantly predict job satisfaction and mental health in this sample of Australian psychiatric nurses. Furthermore, social support was shown to be an important component of the Job Strain Model. PMID- 9871825 TI - Helping methods used by nurses in a psychiatric hospital environment. AB - This qualitative study aims to describe the helping methods used by psychiatric nurses in a hospital environment from as evidenced by video observations and nurses' and patients' descriptions. The data, which were collected by videotaping different nursing situations and by interviewing the nurses and patients afterwards, consisted of 520 pages of written text. A total of 569 utterances were extracted from the text to describe the helping methods used in psychiatric nursing. Each utterance constituted a classification unit. Deductive content analysis techniques were used to analyze the data. The results obtained from the nurses' and patients' descriptions and the videotaped episodes showed that the most commonly used helping methods were of the confirmatory kind (50%), while educational methods came second (37%) and catalytic methods third (13%). PMID- 9871826 TI - Coping and family relationships in stress resistance: a study of job satisfaction of nurses in Singapore. AB - This study examined the role of coping strategies and family relationships in mitigating the negative effect of work stress on nurses. The findings indicated that nurses who adapted to work stress with high job satisfaction were more inclined to adopt approach coping methods (problem orientation, ability enhancement, and change of perspective) than those who reported low job satisfaction under high work stress. The stress-resistant nurses were also distinguished by a less frequent use of defensive or avoidance coping in handling of their emotional reactions to stress. They perceived greater family support than did the distressed nurses. Implications of the findings and limitation of the study were discussed. PMID- 9871827 TI - The seductions of history: sexual trauma in Freud's theory and historiography. AB - The rise of Freud's seduction theory can be illuminated by contextualising aspects of this theory with regard to nineteenth-century trauma theory, germ theory, degeneration theory and sexology. The author argues that Freud, by putting forth his seduction theory, defined himself within a number of registers that organised one's identity within the medical community in late nineteenth century Vienna. Freud's first historical accounts of his rejection of the seduction theory also point to factors that were of great importance to his medical community, issues of epidemiology. However, in 1914 Freud wrote a different history of these events for a different community. No longer a doctor struggling for recognition from the Viennese medical establishment, Freud was now the leader of the psychoanalytic movement, writing a history of the movement with the defections of Adler and Jung in mind. Hence, the author argues that Freud's 1914 version of the fall of the seduction theory served to tie the rejection of the seduction theory to the acceptance of libido theory, a sexual libido theory that Adler and Jung had rebelled against. PMID- 9871828 TI - The splitting of the New York Psychoanalytic Society and the construction of psychoanalytic authority. AB - The splitting apart of the New York Psychoanalytic Society, which began in 1941 with the expulsion of Karen Horney, is seen in the context of a concerted effort to establish the professional authority of psychoanalysis on unimpeachable foundations. On the one hand, a core group of young psychiatrists trained abroad, led by Kubie, sought to establish strict standards of training along orthodox lines; following in the footsteps of medical reformers who had established the professional authority of medicine with strict standards for training and credentialling, they opposed their more 'lax' elders as well as revisionists such as Horney and Rado. On the other hand, the refugee analysts fleeing Hitler sought to establish the security and conditions of orthodoxy they had enjoyed abroad, though they were forced to compromise on the issue of lay analysis. Together they expelled the deviants and sought to establish the professional authority of psychoanalysis on the twin foundations of medical standards and Freudian orthodoxy. Anxieties following the death of Freud contributed to this dynamic process. But contradictions between the conflicting demands of these two sources of authority undermined the stability of this alliance and, eventually, contributed to its collapse. PMID- 9871829 TI - On violence. A preliminary communication. AB - The author argues that although aggression is a central feature of psychoanalytic theory and clinical thinking, little attention has been given by psychoanalysts to the matter of violence. Manifest violence is heterogenous and an adequate understanding of it would involve a multi-disciplinary approach. The author focuses on consideration of a psychoanalytic contribution to this understanding. From this point of view all acts of violence may be characterised as one of two types, or a combination of them, namely self-preservative violence and sado masochistic violence. These are characterised and differentiated; and the relationship between the two is considered, while use is made of clinical material. The issue of 'violence in the transference' is illustrated and briefly discussed. PMID- 9871830 TI - Non-interpretive mechanisms in psychoanalytic therapy. The 'something more' than interpretation. The Process of Change Study Group. AB - It is by now generally accepted that something more than interpretation is necessary to bring about therapeutic change. Using an approach based on recent studies of mother-infant interaction and non-linear dynamic systems and their relation to theories of mind, the authors propose that the something more resides in interactional intersubjective process that give rise to what they will call 'implicit relational knowing'. This relational procedural domain is intrapsychically distinct from the symbolic domain. In the analytic relationship it comprises intersubjective moments occurring between patient and analyst that can create new organisations in, or reorganise not only the relationship between the interactants, but more importantly the patient's implicit procedural knowledge, his ways of being with others. The distinct qualities and consequences of these moments (now moments, 'moments of meeting') are modelled and discussed in terms of a sequencing process that they call moving along. Conceptions of the shared implicit relationship, transference and countertransference are discussed within the parameters of this perspective, which is distinguished from other relational theories and self-psychology. In sum, powerful therapeutic action occurs within implicit relational knowledge. They propose that much of what is observed to be lasting therapeutic effect results from such changes in this intersubjective relational domain. PMID- 9871831 TI - A psychoanalytic concept of metaphor. AB - Metaphor is defined as seeing something in terms of something else. Psychoanalysis sees the present in terms of the past and the past in terms of the present. It therefore relates past and present metaphorically to each other, which implies that both metaphor parts change each other. This view entails a broadening of the linguistic metaphor concept towards a supralinguistic, here psychoanalytic, one: metaphor is not only a phenomenon of language but of mentation; its units are not only words but psychodynamics. Familiar concepts of psychoanalytic theory are recast in the light of this psychoanalytic concept of metaphor. Symptoms are seen as the analysand's damaged ability to metaphorise past and present. Using interpretation the analyst helps the analysand to restore metaphorical processes that have been interrupted in their flow from the past to the present and from the present to the past. Metaphor restores the polysemy that is lost in neurotic suffering. The metaphorical processes within psychoanalysis are seen as common to all psychoanalytic schools that focus on the analysis of transference. The increasing attention the concept of metaphor is receiving within psychoanalysis is seen as related to a trend in today's science to more from objective categories to experiential ones. PMID- 9871832 TI - Transience: its beauty and danger. AB - The author argues that what he terms transition and what he will call transience go together quite often, but that analysis usually discuss transition rather than transience. In the first half of this paper, he examines Ukiyo-e and Japanese myths and folk tales, so as to introduce the Japanese concept of transience and its pathological connotation. In the second half, he quotes two treatments of self-destructive patients to show the importance of understanding the negative and positive aspects of transience in psychoanalytical terms. He concludes that masochistic identification with the transitory figure(s) can take place along with limitless accumulation of debt, causing everything, including one's own self, to be felt as transient. He points out that a Japanese word 'Arigatou', which is almost equivalent to 'thank you' in English, literally means 'difficult to exist', and thus the importance in Japan of appreciating the transience of valuable things. He considers that while readers may be reminded of Winnicott's discussion of transitional objects, transition is a phenomenological description of movement, while transience is mainly an emotional state, or one of painful feeling. Transience, he believes, is not only a feature of Japanese clinical phenomena but also universal. PMID- 9871833 TI - Some thoughts on the essence of the tragic. AB - An attempt is made to define the essence of the tragic through an examination of Euripides' 'The Bacchae', a tragedy that deals with the origins of tragedy itself. The action here culminates in the dismemberment of Pentheus by his mother. It is proposed that the tragic may be related to the earliest phases of differentiation of the subject as a separate entity breaking off from the original mother-infant unit. Tragedy, in this view, could be regarded as the enactment of a primal phantasy of the birth of the 'I' as the result of an archaic act of violence. The process of mourning for the loss of the original unity is central to this development. Pentheus' tragic flaw consists in his repudiation of contradictory dualities and his inability to mourn. The integrative function of 'Logos', both in tragedy and in the analytic process, is underlined. It is suggested that 'Logos' aims to generate meaning not by eliminating contradiction but by embodying the foundational human paradox. PMID- 9871834 TI - Who's afraid of psychoanalytic research? Analysts' attitudes towards reading clinical versus empirical research papers. AB - The authors point out that psychoanalytic research papers are cited with less frequency than clinical papers, and, presumably, are read with less frequency. Results from two sets of questionnaires from psychoanalysts indicate that a majority of analysts report high levels of conviction in the rationales and techniques in their clinical work. However, analysts with higher degrees of conviction read fewer research papers than analysts with lower degrees of conviction. The authors speculate that analysts with higher degrees of conviction may have an underlying sense of uncertainty about their analytic work. Their uncertainty may generate concerns that research may raise questions and doubts about their rationales and techniques, and, consequently, they have little interest in empirical psychoanalytic research. Such an attitude would be understandable because analysts sense or explicitly believe that confidence in their work is an important, perhaps essential, element in the mutative effects of treatment, and must be maintained and protected. The authors believe that clinical and research approaches have each contributed to the development of psychoanalysis and that both need to be used. PMID- 9871835 TI - Consciousness: a neuroscience perspective. PMID- 9871836 TI - Biology and sexuality. PMID- 9871837 TI - Review of the psychoanalytic theory of sexuality. PMID- 9871838 TI - Sexuality and gender identity. PMID- 9871839 TI - 'Transitions in psychic structures in the light of deterministic chaos theory'. PMID- 9871840 TI - Sexuality in the age of AIDS. PMID- 9871841 TI - ECT and clozapine in schizophrenia. PMID- 9871842 TI - The relative efficiency of altering pulse frequency or train duration when determining seizure threshold. AB - In 12 depressed inpatients referred for bilateral electroconvulsive therapy (ECT), each patient was titrated at the first treatment session by using an ascending method-of-limits procedure with a step-wise increase in pulse frequency (frequency titration) or train duration (duration titration). At the second treatment session, seizure threshold was redetermined by using the method (frequency or duration titration) not used at the first treatment. Frequency or duration was maintained at the lowest level when the other parameter was titrated. Seizure threshold was significantly lower with duration titration (mean, 90 mC; SD, 27.3) than frequency titration (mean, 114 mC; SD, 35.6; p = 0.03). On average, patients in the duration-titration group required 1.2 (SD, 0.6) subconvulsive stimulations before a seizure was elicited, and patients in the frequency-titration group required 1.7 (SD, 0.9) subconvulsive stimulations before a seizure was elicited, a nonsignificant difference. These findings suggest that to elicit a seizure during ECT, increasing train duration may be slightly more efficient than increasing frequency. Basic and other clinical research findings indicate that increasing pulse width may be an inefficient way to elicit a seizure. Therefore the following sequence in the determination of seizure threshold is worth considering when using dose-titration or related techniques: the train duration should be increased first before increasing pulse frequency, and the decision to increase pulse width should be reserved for patients who do not seize at the maximal duration and frequency settings. Further empiric research is needed to establish the utility of this approach. PMID- 9871843 TI - Effects of TRH administration on orientation time and recall after ECT. AB - We investigated the effect of thyrotropin-releasing hormone (TRH) on orientation time and recall, in nine depressed female inpatients undergoing electroconvulsive therapy (ECT). In a balanced order crossover design, an intravenous bolus of 0.4 mg TRH or placebo was administered 20 min before ECT in the first two sessions. Orientation time and retrograde and anterograde components of the memory dysfunction, immediately and 24 h later, were assessed. Administration of TRH did not influence orientation time, word recall, or immediate short story recall compared with placebo. We did find, however, an improvement in the number of short story items recalled after 24 h when patients were given TRH compared with placebo. This indicates that TRH may have a protective role against the specific negative effect of ECT on delayed recall. PMID- 9871844 TI - Lithium and maintenance electroconvulsive therapy. AB - This clinical report describes the successful use of a combination of lithium carbonate and maintenance electroconvulsive therapy (ECT) in the treatment of severe bipolar depression. The patient was initially taken off the lithium and given a course of ECT with remarkable improvement in symptoms. He subsequently underwent maintenance ECT, during which lithium was restarted without any side effects. Rating scales were used to assess memory, depression, and mental status. This case suggests that the overlap of the two treatments may be especially beneficial when the plan is to taper maintenance ECT while continuing pharmacotherapy. PMID- 9871845 TI - Labetalol does not lengthen asystole during electroconvulsive therapy. AB - Labetalol, a combined alpha- and beta-adrenergic blocker is often used to attenuate the transient increases in heart rate and blood pressure that accompany electroconvulsive therapy (ECT). It has been suggested that labetalol should not be administered during ECT without the protection provided by anticholinergic medications, because of its potential severe bradycardic effects. We present our experience with 32 patients from all age groups who received labetalol without anticholinergic treatment during ECT. None of the patients demonstrated adverse bradycardic effects. We conclude that administration of labetalol during ECT does not routinely require premedication with anticholinergic drugs and does not lengthen asystole. PMID- 9871846 TI - Electroconvulsive therapy in the first trimester of pregnancy. AB - We present the case of a young woman, 8 weeks pregnant, who had a spontaneous abortion after her third electroconvulsive therapy (ECT) session. We review the literature on ECT in the first trimester of pregnancy. PMID- 9871847 TI - Reimbursement and documentation issues in an ambulatory ECT program. AB - In this era of decreased health resources and the strong influence of third-party payers, the financial advantages of ambulatory electroconvulsive therapy (ECT) should be well recognized by all concerned parties. However, to a large degree, third-party payers are only minimally aware of the availability and potential cost savings associated with this treatment modality. Also, documentation has been problematic, because reviewers unfamiliar with ECT have not been able to find the necessary data even when they are present in the chart. This article illustrates some of these problems and how they have affected our ambulatory ECT program. PMID- 9871848 TI - Formula method for stimulus setting in bilateral electroconvulsive therapy: relevance of age. AB - Seizure thresholds were determined by titration in consecutive electroconvulsive therapy (ECT) patients at the first (n = 146) and sixth (N = 83) ECT sessions. Equations to predict the threshold at the first and sixth ECT were computed from these data using a stepwise linear regression model. These equations were tested prospectively at the first ECT (n = 48) and sixth ECT (n = 26) sessions. Stimulus dose derived from the corresponding equations yielded adequate seizure (successful) in 82% and 84% of patients, respectively. Predictions based on age alone (disregarding two other significant variables--illness severity and inion nasion distance) were estimated at these two ECT sessions. This would not appreciably compromise the success rate. The "formula" method using age alone may therefore be used in routine clinical practice. PMID- 9871849 TI - ECT in the presence of intracranial aneurysm. AB - Two patients with known intracranial aneurysms (ICA) received electroconvulsive therapy (ECT). The first patient received ECT 4 years after the clipping of a right middle cerebral artery aneurysm. Treatment with esmolol failed to completely prevent an increase in blood pressure, but she did well nonetheless. The second patient underwent clipping of a basilar artery aneurysm 11 years before ECT. At the time of ECT, angiography showed a right paraophthalmic artery aneurysm. He received esmolol before each ECT, and, like the first patient, still had significant peri-ictal increases in his blood pressure, but otherwise tolerated the procedure well. Our experience and previously published literature indicate that even with pretreatment, patients often sustain significant increases in their blood pressure. However, this has not been accompanied by any morbidity. Possible guidelines for the management of ICA in association with ECT are discussed. PMID- 9871850 TI - Sumatriptan for post-ECT headache. AB - A 27-year-old woman receiving electroconvulsive therapy (ECT) for the treatment of depression developed post-ECT headaches refractory to treatment with acetaminophen and various nonsteroidal antiinflammatory drugs. The patient was then treated successfully with oral sumatriptan. This case suggests that sumatriptan may be a useful treatment of post-ECT headaches that are unresponsive to conventional analgesics. PMID- 9871851 TI - ECT in dissociative identity disorder and comorbid depression. AB - Dissociative identity disorder (DID), previously named multiple personality disorder, is a diagnosis often complicated by comorbid major depression. We report on four cases of DID associated with severe self-destructive behavior and comorbid major depression treated with electroconvulsive therapy (ECT). In three of the patients, ECT appeared to be helpful in treating the comorbid depression without adversely affecting the DID. The potential risks of using ECT in patients with DID are reviewed. PMID- 9871853 TI - Pre-ECT cardiology consultation. PMID- 9871852 TI - Concurrent administration of clozapine and ECT: a successful therapeutic strategy for a patient with treatment-resistant schizophrenia. AB - A male patient with a 4-year history of schizophrenic disorder, treated in the past with several conventional neuroleptics, atypical antipsychotics, as well as adjunctive therapies, was hospitalized because of treatment resistance/intolerance to pharmacotherapy. During 60 weeks of hospitalization, the patient was treated with conventional neuroleptics or clozapine monotherapy and with concurrent bilateral electroconvulsive therapy (ECT). The patient showed inadequate response to conventional neuroleptics or clozapine without ECT. Concurrent administration of pimozide and ECT resulted in a brief period of improvement. However, concurrent administration of clozapine and ECT resulted in substantial improvement. Our observation suggests that combination treatment with clozapine and ECT can be safe and effective. PMID- 9871854 TI - Theophylline and ECT. PMID- 9871855 TI - Safe use of ECT with an intracranial aneurysm. PMID- 9871856 TI - The tale of normal-tension glaucoma. PMID- 9871857 TI - The definition of normal-tension glaucoma. AB - PURPOSE: To evaluate the consistency of the definition of normal-tension glaucoma (NTG). METHODS: All articles about NTG published between 1973 and 1997 from three clinical journals of ophthalmology were identified by a literature search. They were reviewed to determine criteria used to define NTG, including optic nerve assessment and intraocular pressure (IOP) requirements. RESULTS: There were 63 articles identified that defined the disease entity NTG, including 30 published between 1973 and 1992 and 33 published between 1993 and 1997. Maximum IOP values acceptable for designation as NTG ranged from 17 mmHg to 26 mmHg. A significantly greater proportion of studies published during the last 5 years required maximum IOP values < or = 21 mmHg. Inclusion criteria based on structural characteristics of the glaucomatous optic disc or on the glaucomatous visual field were present in 55 (87%) and 56 (89%) articles, respectively. Of the seven articles that listed specific features classifying a disc as glaucomatous, seven (100%) were published within the last 5 years. Of the 13 articles that listed specific features classifying a field as glaucomatous, 9 (69%) were published within the last 5 years. CONCLUSION: The criteria used to define NTG during the last 25 years have been highly variable. However, publications during the last 5 years have been more likely to require a maximum IOP < or = 21 mmHg and to describe specific features of the glaucomatous optic disc and visual field. PMID- 9871858 TI - Clinical factors associated with progression of normal-tension glaucoma. AB - PURPOSE: To investigate clinical factors associated with the progression of visual field loss in normal-tension glaucoma (NTG). METHODS: One hundred and ten patients with NTG whose cases were followed for more than 2 years were retrospectively analyzed with the Kaplan-Meier life table method and the Cox proportional hazards model. Several clinical factors were investigated to find a possible association with progression of glaucomatous visual field defined by two different definitions: one by mean deviation change and the other by pointwise comparison. RESULTS: The Cox proportional hazards model indicated that change in the visual field was significantly associated with treatment with calcium channel blockers, recovery rate from a cold recovery test, systolic blood pressure, disc hemorrhage, corrected pattern standard deviation, mean deviation, and fluctuation of intraocular pressure (IOP) at a 24-hour phasing. The probability of nonprogression of visual field loss was significantly higher for patients taking calcium channel blockers than for control subjects by either definition of visual field progression. CONCLUSION: Factors other than IOP, including the use of calcium channel blockers, are associated with the outcome of visual field loss in NTG. PMID- 9871859 TI - Detection of optic disc changes with Glaucoma-Scope probability maps. AB - PURPOSE: To test whether a statistical method using a probability map could detect true changes in optic disc topography. METHODS: The average of three Glaucoma-Scope images (Ophthalmic Imaging Systems were used for analysis at each of two sessions. A Glaucoma-Scope probability map was constructed for each eye using statistical methods. The proportion of topographic locations with p values less than 0.05 on a modified two-sample t test (p-proportion) and the difference in the mean position of the disc (MPD) from two imaging sessions were calculated. Two pairs of stereoscopic disc photographs for 43 eyes with longitudinal follow up were evaluated for change by four experienced glaucoma specialists masked to patient clinical information. Clinical change was considered to have occurred when the assessments of at least three of the four specialists were agreed on. The cutoff values for p-proportion and change in MPD that provided 95% specificity were calculated using a separate sample of 69 subjects who had serial images taken at two separate sessions on the same day, and thus showed no clinical change in the optic disc. RESULTS: The cutoff values of 95% specificity for the p-proportion and the change in MPD were 18% and 25.1 microns, respectively. Of 43 eyes with longitudinal follow-up, 14 showed definite clinical change. Sensitivity of the p-proportion and change in MPD for detecting this change was 100% and 85.7%, respectively. For all 43 patients with longitudinal follow-up, the percent change in intraocular pressure (IOP) correlated strongly with both the p-proportion and the change in MPD. CONCLUSION: Using data obtained with the Glaucoma-Scope, a statistical method based on probability mapping can be used to detect true changes in disc topography. The p-proportion was more sensitive than change in MPD in detecting clinical change in the study eyes. This statistical methodology may also be applicable for interpretation of data obtained with other optic disc analyzers. PMID- 9871860 TI - The effects of unoprostone isopropyl 0.12% and timolol maleate 0.5% on diurnal intraocular pressure. AB - PURPOSE: To compare the effect of unoprostone isopropyl 0.12% to that of timolol maleate 0.5% solution given twice daily on the diurnal curve of intraocular pressure (IOP) in patients with primary open-angle glaucoma or ocular hypertension. METHODS: In this investigator-masked, single-center, parallel-group comparison, 36 patients with primary open-angle glaucoma or ocular hypertension were randomized in a 2:1 ratio to receive either unoprostone isopropyl 0.12% or placebo/timolol maleate 0.5% solution, respectively. A placebo-controlled diurnal curve on day 0 and active-controlled diurnal curves at weeks 2 and 4 were performed at 0, 2, 4, 6, 8, 10, 12, and 24 hours. At week 2, administration of unoprostone isopropyl twice daily was compared with administration of timolol maleate twice daily. At week 4, administration of unoprostone isopropyl three times daily was compared with administration of timolol maleate twice daily. RESULTS: At the 24-hour 8:00 AM trough at week 2, administration of unoprostone isopropyl twice daily decreased IOP from 23.4 +/- 2.0 mmHg at baseline to 19.3 +/ 4.4 mmHg, and timolol maleate reduced IOP from 24.4 +/- 2.6 mmHg to 17.5 +/- 2.9 mmHg. At the 8:00 AM trough at week 4, unoprostone isopropyl given three times daily produced an IOP of 19.6 +/- 3.3 mmHg and timolol maleate resulted in an IOP of 19.4 +/- 3.0 mmHg. No statistical differences between groups were observed at any time point during either diurnal curve. Safety was similar in the two treatment groups, with no differences between groups in conjunctival hyperemia, anterior segment inflammation, or iris color change. CONCLUSION: Results of this short-term pilot trial indicate that unoprostone isopropyl may be safe and effective in reducing IOP from baseline when given twice or three times daily. PMID- 9871861 TI - Two-year safety study of dorzolamide as monotherapy and with timolol and pilocarpine. Dorzolamide Safety Study Group. AB - PURPOSE: To evaluate the safety of open-label 2.0% dorzolamide as monotherapy and when used with timolol and/or pilocarpine for as long as 2 years. METHODS: The safety of dorzolamide was evaluated in patients with open-angle glaucoma or ocular hypertension over a 2-year period. The incidence of the most common drug related adverse experiences in the first year was compared with that in the second year using McNemar's test. The ocular hypotensive effect of dorzolamide as monotherapy and with adjunctive therapy was assessed using percent change in intraocular pressure (IOP) from baseline. RESULTS: Of the 304 patients enrolled, 164 (53.9%) continued to receive dorzolamide as monotherapy for 2 years and 140 (46.1%) required add-on therapy. Add-on therapy was initiated by month 6 in 112 of these 140 patients (80%). Of the 304 patients, 202 (66.4%) completed 2 years of therapy. Of the patients who received dorzolamide as monotherapy, drug-related adverse events occurred more frequently during the first year (29.7%) than the second year (13.8%), and the most common ocular drug-related adverse events included conjunctivitis, burning/stinging eye, follicular conjunctivitis, and eyelid edema. After 2 years of therapy, the mean percent decrease in peak IOP was 22.8% for patients receiving dorzolamide monotherapy and 31.2% to 36.0% for patients receiving add-on therapy. CONCLUSION: Dorzolamide was generally well tolerated for up to 2 years as monotherapy and when used with timolol and/or pilocarpine. Drug-related adverse events were less frequent during the second year of monotherapy than during the first year. Most patients who required add-on therapy did so within the first 6 months of initiating dorzolamide therapy. PMID- 9871862 TI - Efficacy and safety of timolol solution once daily versus timolol gel in treating elevated intraocular pressure. AB - PURPOSE: To evaluate the efficacy and safety of timolol hemihydrate once daily versus timolol maleate gel forming solution once daily in patients with primary open-angle glaucoma or ocular hypertension. METHODS: We prospectively randomized patients with primary open-angle glaucoma or ocular hypertension to receive either timolol hemihydrate 0.5% solution or timolol maleate gel forming solution 0.5% every morning. The primary efficacy variable was the 8:00 AM trough intraocular pressure (IOP) 24 hours after administration. RESULTS: Three months after initiation of therapy, baseline IOP had decreased from 23.6 +/- 1.9 mmHg to 18.3 +/- 2.8 mmHg in the group taking timolol hemihydrate (n = 22) and from 23.7 +/- 2.2 mmHg to 18.4 +/- 3.1 mmHg in the group receiving timolol maleate gel (n = 21) at the 24-hour trough level. This was not a significant difference between groups at 3 months. Also, no difference was observed between groups in the 2-hour post instillation IOP. Visual acuity was decreased in the group receiving timolol maleate gel compared with those receiving timolol hemihydrate one minute after instillation of study medicine at month 3. Otherwise, ocular and systemic safety were similar between groups. No differences between groups in cardiac pulse or systolic and diastolic blood pressure were observed. CONCLUSION: Timolol hemihydrate 0.5% solution given once a day appears to be as efficacious and safe in decreasing IOP as timolol maleate gel 0.5% given once a day. PMID- 9871863 TI - Age and increased incidence of "forward bowing" of the iris in normal eyes. AB - PURPOSE AND METHODS: The incidence of forward bowing of the iris, width of the iridocorneal angle, and distance between the apex of the angle and the scleral spur were studied using high-frequency (50 MHz) ultrasound B-scan images in 90 normal eyes from Japanese volunteers. RESULTS: The incidence of forward bowing of the iris, defined as a 120-micron anterior shifting of the posterior profile of the iris, increased from 0% in subjects under 31 years old to 50% in subjects 71 years of age and older. The iridocorneal angle was significantly narrower and the distance between the apex of the angle and the scleral spur was significantly less in eyes with forward bowing. Of the risk factors (age, axial length, sex, and corneal curvature), age (p = 0.0005) was the most significant risk factor for forward bowing of the iris. CONCLUSION: Forward bowing of the iris is not uncommon in the elderly. Half of the Japanese subjects in this study 71 years of age and older had this condition, even though they had no optic nerve atrophy or elevation in intraocular pressure (IOP) at the time of examination. PMID- 9871864 TI - Pulsatile ocular blood flow and intraocular pressure during retrobulbar injection of lignocaine: influence of additives. AB - PURPOSE: To assess the influence of adrenaline and hyaluronidase on pulsatile ocular blood flow and intraocular pressure changes during retrobulbar anesthesia. METHOD: Patients undergoing cataract surgery under local anesthesia received a 5 mL retrobulbar injection of lignocaine with either adrenaline, hyaluronidase, or no addition. Continuous pneumotonometry was performed using the OBF tonograph (OBF Labs, Chippenham, UK), which automatically derives pulsatile ocular blood flow values. Measurements were taken before and one minute after injection, and changes were compared between groups to evaluate the effects of changing the constituents of the injection. RESULTS: There was a marked decrease in pulsatile ocular blood flow during injection in all groups. In the group receiving lignocaine with hyaluronidase, the decrease in pulsatile ocular blood flow (mean 141 microL/min) was significantly less than in either the group receiving lignocaine and adrenaline (mean -249 microL/min) or the group receiving lignocaine alone (mean -245 microL/min). There was no significant change in intraocular pressure in any group. CONCLUSION: Hyaluronidase appears to lessen the reduction in pulsatile ocular blood flow. Injection volumes of 5 mL need not cause significant intraocular pressure change. PMID- 9871865 TI - Use of autologous Tenon's capsule and scleral patch grafts for repair of excessively draining fistulas with leaking filtering blebs. AB - PURPOSE: To report the use of autologous Tenon's and partial-thickness scleral tissues for patch graft revisions of excessively draining fistulas associated with leaking filtering blebs. METHODS: Appropriate sized pieces of autologous Tenon's tissue and partial-thickness scleral tissue were dissected near the surgical sites and used as patch grafts to effectively repair excessively draining fistulas with leaking filtering blebs. RESULTS: Both Tenon's and partial thickness scleral patch grafts were useful in adequately closing the excessively draining fistulas. CONCLUSION: Autologous Tenon's and scleral tissue patch grafts appear to be safe and effective for the repair of excessively draining fistulas, and are useful when patch grafts are unexpectedly needed. PMID- 9871866 TI - Descriptive information of topographic parameters computed at the optic nerve head with the Heidelberg retina tomograph. AB - PURPOSE: To analyze the frequency distribution and descriptive information of topometric data obtained with the Heidelberg Retina Tomograph (HRT) in a normal population. METHODS: Topographic measurements of the optic disc were acquired and evaluated using the HRT in 225 subjects between 12 and 90 years of age. After randomly selecting one eye per subject, the frequency distributions, mean values, minima, maxima, and first, fifth, fiftieth, ninety-fifth, and ninety-ninth percentiles were evaluated for topographic parameters computed by the HRT. The influence of age, intraocular pressure (IOP), disc size, and disc shape on optic disc topometric data was analyzed. A principal component analysis of the topometric parameters was performed. The frequency distributions, mean values, minima, maxima, and first, fifth, fiftieth, ninety-fifth, and ninety-ninth percentiles of the interocular difference in topographic parameters were evaluated. RESULTS: All topographic parameters showed a unimodal but not necessarily normal distribution. None of the parameters showed a relevant correlation with age, IOP (in the normal range), and overall shape of the anterior optic nerve, but a few parameters showed a clinically significant correlation with disc size. A principal component analysis identified four relevant factors (optic nerve cup, retinal nerve fiber layer, optic disc size, and optic nerve cup shape) in the entire data set of optic nerve topometric data. The absolute value of all interocular differences in topographic parameters showed an asymmetric but unimodal distribution. CONCLUSION: The mathematical description of the optic nerve cup shape provides information on optic nerve head topography independently from cupping, nerve fiber layer thickness, and disc size. Potentially, quantification of further aspects in optic nerve head topography might improve the discriminatory power of computerized quantitative optic nerve head analysis. PMID- 9871867 TI - Traumatic cyclodialysis cleft. PMID- 9871868 TI - New conceptual approaches for pharmacological neuroprotection in glaucomatous neuronal degeneration. PMID- 9871869 TI - Fitting contact lenses in eyes with filtering blebs. PMID- 9871870 TI - Situation with glaucoma and the state of mind in the glaucoma community. PMID- 9871871 TI - Internet usage and potential impact for acute care hospitals: survey in the United States. AB - These survey results are from a national survey of acute care hospitals. A random sample of 813 hospitals was selected with 115 responding and 33 incorrect addresses resulting in a 15% response rate. The purpose of the study was to measure the extent of information systems integration in the financial, medical, and administrative systems of the hospitals. Internet usage including homepages and advertising was measured. Other selected telecommunication applications are analyzed. As demonstration projects from the literature are compared to the survey results, the potential for hospitals is tremendous. Resulting cost savings could be equally impressive. This information will provide a benchmark for hospitals to determine their position relative to Internet technology and to set goals. PMID- 9871872 TI - Impact of information systems on acute care hospitals: results from a survey in the United States. AB - These results are from a survey of acute care hospitals in the United States. A random sample of 813 hospitals was selected with 115 responding and 33 incorrect addresses resulting in a 15% response rate. The purpose of the study was to measure the impact and future potential of information systems integration in the financial, medical, and administrative systems of the hospitals. Impact of Information Systems is measured in many ways. Questions about employee moral, reductions in employees, goals being met and overall satisfaction with the systems are asked and results analyzed. This information will provide a benchmark for hospitals to determine their information systems position and technology transfer planning. These analyses will lead to setting goals. PMID- 9871873 TI - Information systems' approaches and designs and facility information: survey of acute care hospitals in the United States. AB - These survey results are based on a national survey of acute care hospitals. A random sample of 813 hospitals was selected with 115 responding and 33 incorrect address returns resulting in a 15% response rate. The purpose of the study was to measure the extent of information systems integration in the financial, medical, and administrative systems of the hospitals. The rate of adoption of information technology or technology transfer is discussed. Facility demographics are presented. Characteristics of information systems' approaches and designs are reviewed and their implications discussed. This information will provide benchmarks for hospitals to determine their information systems position and to set goals. PMID- 9871875 TI - Smart cards: a specific application in the hospital. AB - Computers have the ability to process and access tremendous amounts of information in our daily lives. But, now, individuals have this ability by carrying a smart card in their own wallets. These cards provide us the versatility, power, and security of computers. This study begins with a short description of smart cards and their advantages. Then, an electronic circuit that is designed for healthcare application in hospitals is introduced. This circuit functions as a smart card holder identifier, access controller for hospital doors and also can be used as a smart card reader/writer. Design steps of this electronic circuit, operation principles, serial communication with P.C., and the software are examined. Finally a complete access control network for hospital doors that functions with smart cards is discussed. PMID- 9871874 TI - Decision-making with and without information technology in acute care hospitals: survey in the United States. AB - These survey results are from a national survey of acute care hospitals. A random sample of 813 hospitals was selected with 115 responding and 33 incorrect addresses resulting in a 15% response rate. The purpose of the study was to measure the extent of information systems integration in the financial, medical, and administrative systems of the hospitals. Decision making with and without information technology is explored based upon the survey data. The results indicate why and how meetings are held. Necessary changes in the decision-making environment are identified for decision making utilizing information technology to be successful. These results will provide a benchmark for hospitals to determine their technology transfer position and to set goals for computer assisted decision-making. PMID- 9871876 TI - Problem based learning as a curricular strategy for medical undergraduates: a tutor's experience. AB - This article focuses on the experiences and role of a tutor while conducting a problem based learning (PBL) exercise to facilitate the learning of biochemistry and molecular biology. A case presentation of sickle cell anemia was used to frame a module. The objective of this experimental exercise was to assess the suitability and effectiveness of problem based learning in the first year of undergraduate medical course and to practice self-assessment by tutors in this new role of a facilitator of learning through small group discussion. A tutor during such a learning session encouraged the students to apply their reasoning to analyze the problem and to develop self-directed learning skills in acquiring the knowledge appropriate to their perceived needs to work on a problem. The tutors ensured that they apply this knowledge in their work with other similar problems that they would encounter later in life. PMID- 9871877 TI - The computer based patient record: a strategic issue in process innovation. AB - Reengineering of the workplace through Information Technology is an important strategic issue for today's hospitals. The computer-based patient record (CPR) is one technology that has the potential to profoundly modify the work routines of the care unit. This study investigates a CPR project aimed at allowing physicians and nurses to work in a completely electronic environment. The focus of our analysis was the patient nursing care process. The rationale behind the introduction of this technology was based on its alleged capability to both enhance quality of care and control costs. This is done by better managing the flow of information within the organization and by introducing mechanisms such as the timeless and spaceless organization of the work place, de-localization, and automation of work processes. The present case study analyzed the implementation of a large CPR project ($45 million U.S.) conducted in four hospitals in joint venture with two computer firms. The computerized system had to be withdrawn because of boycotts from both the medical and nursing personnel. User-resistance was not the problem. Despite its failure, this project was a good opportunity to understand better the intricate complexity of introducing technology in professional work where the usefulness of information is short lived and where it is difficult to predetermine the relevancy of information. Profound misconceptions in achieving a tighter fit (synchronization) between care processes and information processes were the main problems. PMID- 9871878 TI - Cadmium and air showers. PMID- 9871879 TI - Reproductive hormone profile among pesticide factory workers. AB - Serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone levels, as well as urinary levels of FSH, LH, and E1C, a metabolite of testosterone, were measured to investigate the adverse reproductive effects of organophosphate pesticides among Chinese factory workers who were occupationally exposed to ethylparathion and methamidophos. Thirty-four exposed workers were randomly chosen and recruited from a large pesticide factory, and 44 unexposed workers were selected from a nearby textile factory. A quantitative pesticide exposure assessment was performed among a subset of the exposed and unexposed workers. Information on potential confounders was collected in an interview. A single blood sample was collected at the end of a work shift, when each subject also donated a semen sample. Three first-voided urine samples were collected from each worker on 3 consecutive days. Urinary p-nitrophenol level at 1 hour after the work shift correlated with serum (r = 0.71, P < 0.01) and urinary (r = 0.51, P = 0.04) FSH levels. Stratifying by the subjects' exposure status, we found a significant negative correlation among the exposed group between urinary FSH level and sperm count (r = -0.61, P < 0.01) and between urinary FSH level and sperm concentration (r = -0.53, P = 0.03). Pesticide exposure alone was significantly associated with serum LH level (beta [coefficient of exposure effect] = 0.79; 95% confidence interval [CI] = 0.42, 1.16) but not with serum FSH or testosterone or with any urinary hormone levels. With adjustment for age, rotating shift work, current cigarette smoking, and current alcohol consumption, exposure significantly increased the serum LH level by 1.1 mIU/mL (95% CI = 0.34, 1.82). Meanwhile, the serum FSH level was slightly elevated (beta [coefficient of exposure effect] = 1.38; 95% CI = -0.09, 2.85) and the serum testosterone level was decreased (beta = -55.13; 95% CI = -147.24, 37) with increased pesticide exposure. Age and rotating shift work appeared to act as confounders. We conclude that organophosphate pesticides have a small effect on male reproductive hormones, suggestive of a secondary hormonal disturbance after testicular damage. PMID- 9871880 TI - Longitudinal changes in lung function associated with aspects of swine confinement exposure. AB - Several aspects of swine-confinement farming appear to be leading to adverse respiratory effects. This study was set up in a longitudinal design to study the association between certain characteristics of farms or the way they are run and a decline in lung function. A cohort of 171 pig farmers was observed for 3 years. Lung function was measured. Exposure to farm characteristics was determined at the start of the observation period, using data from standardized farm surveys and from diaries kept by the participants. Mean decline in lung function was 73 mL/year for forced expiratory volume in 1 second (FEV1) and 55 mL/year for forced vital capacity (FVC). A longitudinal decline in FEV1 was significantly associated with the use of quaternary ammonium compounds as disinfectants (an additional 43 mL/year) and also with the use of an automated dry feeding system (an additional 28 mL/year). The association with the use of wood shavings as bedding material was not statistically significant. The impact of these characteristics in a longitudinal study provides stronger evidence for causal inference than that shown in previous cross-sectional designs. This may be useful in promoting preventive measures. PMID- 9871881 TI - Plasma Asp13-Ki-ras oncoprotein expression in vinyl chloride monomer workers in Taiwan. AB - Vinyl chloride (VC) workers are known to be at risk for development of liver angiosarcoma, a rare tumor. Previously, more than 80% of VC workers with liver angiosarcoma have been found to have an Asp-13 c-Ki-ras oncogene mutation, and more than 50% of VC-exposed workers without liver tumors were found to have Asp13 Ki-ras oncoprotein in their plasma. Some workers in Taiwan had also been exposed to VC, and some have contracted liver tumors. In this study, we used enhanced chemiluminescence Western blotting to detect Asp13-p21-Ki-ras in the sera of VC exposed workers in Taiwan. There were 14 of 113 (12.4%) VC workers positive for the Asp13-Ki-ras oncoprotein in plasma, but 0 of 18 controls were positive. There were 10 of 69 (14.5%) plasma-positives among the more highly exposed (> 1000 ppm months) workers and 4 of 48 (9.1%) plasma-positives among the lesser exposed (< or = 1000 ppm-months). Compared with the unexposed controls, the odds ratios (and 95% confidence intervals [CI]) for plasma-positivity were 4.11 (95% CI = 0.21, 80.4) in the lower-exposed workers and 6.53 (95% CI = 0.37, 116.9) in the higher exposed workers, and there was a linear trend between exposure and plasma positivity (P = 0.073). After adjusting for age and drinking status, the odds ratios (and 95% CIs) were 1.64 (95% CI = 0.17, 15.8), and 2.65 (95% CI = 0.42, 16.8), respectively, and there was a significant linear trend between exposure and plasma-positivity (P = 0.048). In summary, Asp13-Ki-ras oncoprotein can be found in the plasma of VC workers in Taiwan, and a significant dose-response relationship exists between plasma oncoprotein expression and VC exposure. PMID- 9871882 TI - Eruptive cherry angiomas and irritant symptoms after one acute exposure to the glycol ether solvent 2-butoxyethanol. AB - Seven clerical workers were evaluated in 1993, 8 months after exposure to vaporized 2-butoxyethanol (2-BE; also called butyl cellosolve or ethylene glycol monobutyl ether [EGMBE or EGBE]), which had been applied overnight to strip the floor of their file room. At the time of exposure, they had noted intense eye and respiratory irritation, marked dyspnea, nausea, and faintness, suggesting a concentration of 2-BE in the air of 200-300 parts per million (ppm). All seven workers later experienced recurrent eye and respiratory irritation, dry cough, and headache. Four months after the exposure, cherry angiomas began to appear on the arms, trunk, and thighs of six workers, who voiced concerns about the possibility of cancer. Our evaluation found no evidence of hematologic, liver, lung, or renal toxicity, but elevations in the erythrocyte sedimentation rate and blood pressure of each subject were found. Workplace air sampling found no detectable 2-BE, but traces (0.1-0.2 ppm) of formaldehyde were identified. Irritant symptoms abated after the group was moved to a room with better ventilation, and the mild hypertension gradually cleared, but new cherry angiomas have continued to appear 5 years after the acute exposure, as the initial ones persisted. These angiomas occur in healthy persons as they age but in this instance appear to have resulted from a single overexposure to 2-BE. We felt confident in reassuring the workers that they would suffer no serious consequences from this exposure. PMID- 9871883 TI - Temporal trends in coal workers' pneumoconiosis prevalence. Validating the National Coal Study results. AB - Evidence from four successive rounds of the National Study of Coal Workers' Pneumoconiosis indicates diminishing prevalence of coal workers' pneumoconiosis (CWP) from 1969 to 1988. However, methodological inconsistencies across surveys have raised concerns. This study confirms the reported downward trend in CWP prevalence, utilizing a standardized methodological approach. A single team of three x-ray readers using the 1980 International Labour Office classification independently re-evaluated 3143 Appalachian-region cases to derive overall, tenure- and age-specific prevalences. Prevalence of small rounded opacities declined, with 12.7% in Round 1, 11.2% in Round 2, 3.0% in Round 3, and 3.9% in Round 4. These findings support the National Institute for Occupational Safety and Health's recommendation of a reduced exposure limit of 1 mg/m3 because the present coal dust standard does not sufficiently protect miners against adverse health effects over a working lifetime of exposures. PMID- 9871884 TI - Evaluation of a population with occupational exposure to airborne ammonium perchlorate for possible acute or chronic effects on thyroid function. AB - Employees at an ammonium perchlorate production facility in Nevada and a larger control population from the same chemical complex without direct AP exposure were monitored extensively for airborne perchlorate exposure. Single-shift and working lifetime cumulative dose estimates were made using standard breathing-rate estimates and assuming rapid absorption, based upon solubility. Calculated single shift doses ranged from 0.2 to 436 micrograms/kg, with an average of 36 micrograms/kg. Working-lifetime cumulative doses in the higher exposure group ranged from 8,000 to 88,000 micrograms/kg, with an average of 38,000 micrograms/kg. Thyroid profiles, including free thyroxine index and thyroid stimulating hormone level, were obtained both before shift and after shift to assess thyroid-axis perturbation due to single working-shift perchlorate exposure. Thyroid-function data were also analyzed with respect to estimates of cumulative exposure to assess any measurable chronic effects on thyroid gland function. Additionally, standard clinical blood test parameters of liver, kidney, and bone marrow function were evaluated to assess any measurable chronic effects of perchlorate exposure on those organs. Multiple regression was used to assess the effects of exposure variables and demographic variables on organ function parameters. No perchlorate-attributable effects on thyroid, bone marrow, kidney, or liver function were detected. PMID- 9871885 TI - Occupational burns in Washington State, 1989-1993. AB - Occupational burns cause significant morbidity in the United States each year; however, there are few studies that report industries or workplaces where workers are at an increased risk of burn injuries. Washington State's Department of Labor and Industries (L and I) computerized workers' compensation database was used to describe work-related burns over 5 years. From 1989 to 1993, L and I accepted 27,323 claims for occupational burns, 71.4% of them thermal burns and 26.8% chemical burns. The most common sources of injury were cooking oils (14%) and hot water/steam (13%). Workers involved in food preparation or food handling accounted for the highest proportion of injured workers (30%). Industries involved in the smelting, sintering, or refining of ore had the highest rate for thermal burns, with a rate of 15.0 burn injuries per 100 full-time equivalent workers per year, followed by paper, pulp, or wood fiber manufacturing, with a rate of 5.8, then roof work, with a rate of 4.3. Industries involved in hazardous waste landfill clean-up had the highest rate for chemical burns, with a rate of 4.9, followed by portable cleaning and washing, with a rate of 3.5, and paper, pulp, and wood fiber manufacturing, with a rate of 2.6. Further study is needed to identify work practices that result in burn injuries in order to decrease the incidence of this preventable occupational injury. PMID- 9871886 TI - Validity of spot urine samples as a surrogate measure of 24-hour porphyrin excretion rates. Evaluation of diurnal variations in porphyrin, mercury, and creatinine concentrations among subjects with very low occupational mercury exposure. AB - Porphyrin, mercury, and creatinine levels in single-void urine specimens ("spot samples") were compared with calculated 24-hour urine concentrations among 146 (77 male and 69 female) practicing dentists who are participating in an ongoing study of urinary porphyrin changes as a biomarker of mercury body burden. All subjects had urinary mercury concentrations < or = 5 micrograms/L, a level comparable to that of the general US population and which is below that determined to be associated with mercury-induced changes in urinary porphyrin excretion rates. The results confirmed previous findings of no significant diurnal variation in any of the porphyrin levels normally found in urine or in total porphyrin levels combined among males but describe significant diurnal variations in most porphyrin levels as well as concentrations of total porphyrins combined among females. Similarly, no evidence of large diurnal variation in mercury excretion among males was apparent, whereas significant diurnal variation in the mercury excretion rate among females was found. Creatinine adjustment of porphyrin or mercury concentrations had no significant effect on these findings. Moreover, no evidence of diurnal variation in urinary creatinine excretion among either male or female subjects was obtained, despite substantial between-subject variability in this parameter. These results support the view that spot urine samples may be utilized to derive reasonably accurate estimates of 24-hour porphyrin and mercury excretion rates in male subjects. In contrast, time of day appears to be of considerably greater importance when spot samples are utilized as 24-hour estimates of either porphyrin or mercury excretion rates among females. Additionally, time of day may be an important consideration in studies involving serial (repeated) porphyrin or mercury measurements using spot urine samples, irrespective of gender distribution of study subjects. PMID- 9871887 TI - Occupational exposure to HIV: experience at a tertiary care center. AB - We examined our hospital-based occupational health clinic's experience with combination antiretroviral therapy for postexposure prophylaxis for human immunodeficiency virus (HIV). Over a 12-month period, 68 workers started postexposure prophylaxis: 23 with zidovudine and lamivudine and 45 with zidovudine, lamivudine, and indinavir. Fifty-one (75%) of the 68 workers starting postexposure prophylaxis reported one or more side effects. Side effects were more common among those taking three drugs. Many workers failed to complete the recommended 28-day regimen because of the side effects of the various treatments. The estimated mean cost for evaluations, prophylaxis, and monitoring of exposed workers was $669 per reported exposure. In our experience, major challenges in carrying out the current HIV postexposure prophylaxis guidelines include expeditious source testing, improved staff education and prevention measures, and scrupulous monitoring of workers taking combination antiretroviral drugs for postexposure prophylaxis, with consideration of alternate regimens for intolerant workers. PMID- 9871888 TI - Trends in disability duration and cost of workers' compensation low back pain claims (1988-1996). AB - Previously published epidemiologic studies of low back pain (LBP) have reported that the prevalence of low back disability has increased dramatically. These studies based their findings on either the number of disability claims filed, the disability duration, or both. This information was from countries other than the United States or from the US Social Security Disability Insurance data, with findings reported only to the early 1980s. More recent studies of US workers' compensation LBP claims reported a decrease in the incidence rate from the late 1980s to the mid-1990s. No studies have been found that report on the trends of disability duration for workers' compensation LBP claims. This study examined recent trends in the length of disability (LOD) for LBP claims and associated costs, using a large sample of claims from the privately insured US workers' compensation market. LOD and cost information were derived for injuries from 1988, 1990, 1992, 1994, and 1996. For each year, the distributions of LOD and cost were skewed, with the small percentage of claims that lasted more than one year (4.6%-8.8%, depending on the year) accounting for a large percentage of the total disability days (77.6%-90.1%) and cost (64.9%-84.7%). From 1988 to 1996, the average LOD decreased 60.9%, from 156 days to 61 days. The probability of being on disability for a long period of time has decreased over the years. Over the study period, the average cost of a claim decreased 41.4%, while the median cost increased 19.7%. The most influential change in the LOD and cost distributions was a reduction in expensive claims with a long disability duration. The evolution of LOD and cost is also detailed for different disability durations for the study period. PMID- 9871889 TI - Mortality of short-term workers in two international cohorts. AB - The purpose of this study was to compare the pattern of mortality of blue-collar workers employed less and more than 1 year in the man-made vitreous fiber (MMVF) and the reinforced plastic industries, the latter group being exposed to styrene. We conducted an analysis among 21,784 workers with less than 1 year of employment (short-term workers) and 19,117 workers with 1 or more years of employment (long term workers) employed in eight European countries. We conducted analyses based on external as well as internal comparisons. In both cohorts, the standardized mortality ratio for all causes among short-term workers was approximately 40% higher, compared with that for longer-term workers. In internal comparisons, the difference was reduced to 9% in the MMVF cohort and 11% in the styrene cohort. Workers with less than 1 month of employment displayed an increased mortality in both cohorts and in most countries. The increased mortality among short-term workers was not concentrated shortly after they quit employment. In both cohorts, short-term workers had a higher mortality from external causes, while little difference was seen in mortality from ischemic heart disease and malignant neoplasms. Although extra-occupational factors may contribute to increase the mortality of short-term workers and, in particular, of those employed for less than 1 month, the difference observed in analyses adjusted for characteristics of employment suggested a relatively small difference in mortality from most causes. PMID- 9871890 TI - Evolution of a state occupational lead exposure registry: 1986-1996. AB - In the first 11 full years of operation (January 1, 1986, through December 31, 1996) of an adult lead registry in New Jersey, 23,456 reports of occupational lead toxicity (blood lead level > or = 1.21 mumol/L) in 4,011 workers, involving 496 workplaces, were received. The majority of the reports and workers were from the manufacturing and construction industries. Over the 11 years, the annual numbers of reports and workers declined, although the annual numbers of involved workplaces remained stable, as did the number of newly identified workers and workplaces. The decline occurred primarily in the manufacturing industry; the construction industry experienced an increase in reports and reported workers. For all years combined, 36% of reported workers had at least one blood lead level equal to or greater than 1.93 mumol/L, although in the most recent years the percentage dropped overall and in both the manufacturing and construction industries. PMID- 9871891 TI - Race-specific cancer mortality in US firefighters: 1984-1993. AB - A mortality odds ratio (MOR) study of race-specific cancer risk among firefighters was conducted using 1984-1993 death certificate data from 24 states. The Bureau of the Census Index of Industries and Occupations was used to code occupation on death certificates. The overall cancer mortality was slightly elevated among white firefighters (MOR = 1.1; 95% confidence interval [CI] = 1.1 1.2), but the increase in overall cancer mortality among black firefighters was not significant (MOR = 1.2; 95% CI = 0.9-1.5). Only prostate cancer risk was elevated in both groups (whites: MOR = 1.2; 95% CI = 1.0-1.3; blacks: MOR = 1.9; 95% CI = 1.2-3.2). Among white firefighters, elevated site-specific cancer mortality risks were found for the following cancer sites: lip (MOR = 5.9; 95% CI = 1.9-18.3), pancreas (MOR = 1.2; 95% CI = 1.0-1.5), soft tissue sarcoma (MOR = 1.6; 95% CI = 1.0-2.7), melanoma (MOR = 1.4; 95% CI = 1.0-1.9), kidney and renal pelvis (MOR = 1.3; 95% CI = 1.0-1.7), non-Hodgkin's lymphoma (MOR = 1.4; 95% CI = 1.1-1.7), and Hodgkin's disease (MOR = 2.4; 95% CI = 1.4-4.1). We also observed a slightly elevated risk for bronchus and lung cancer (MOR = 1.1; 95% CI = 1.0 1.2). Among black firefighters, excess risks were found for cancers of the brain and central nervous system (MOR = 6.9; 95% CI = 3.0-16.0), colon (MOR = 2.1; 95% CI = 1.1-4.0), and nasopharynx (MOR = 7.6; 95% CI = 1.3-46.4). Future studies are needed to confirm the existence of differential cancer mortality risks among firefighters of different race/ethnic subpopulations. PMID- 9871892 TI - [Parenteral nutrition associated cholestasis in the newborn]. AB - Parenteral nutrition associated cholestasis in preterm infants and newborn children is a frequent and serious disease with an incidence of 23% depended on duration of parenteral nutrition and birthweight. The incidence of liver cirrhosis is 40% when parenteral nutrition is given 74-242 days. The pathogenesis remains unclear. Several predisposing factors are discussed like immaturity, lack of hormonal stimulation by oral feeding, bacterial infection, liver toxicity of aminoacids and their products of photooxidation, lack of taurine, lack of antioxidation substances, hypermanganesaemia and pollution of infusion solutions. Furthermore sepsis during parenteral nutrition seems to multiply the risk of cholestasis. For prevention controlled studies recommend: 1. Early enteral nutrition. 2. The reduction of parenteral amino acids to less than 3 g/kg/d. 3. Light protection for parenteral solutions. 4. Cyclic infusion of parenteral nutrition. 5. The application of antibiotics (metronidazole, gentamicin) during parenteral nutrition. The most important therapeutic intervention is the beginning of oral feeding. Most of the time this leads to a decrease of icterus within two weeks. An icterus persisting longer than 3 weeks should be treated because of the risk of liver cirrhosis. Further therapeutic interventions are: 1. Cholecystokinin, good results in case studies which still has to be verified by a controlled study. 2. Ursodeoxycholic acid, its choleretic effectiveness is verified in several liver diseases by controlled studies, but it is not proven in parenteral nutrition associated cholestasis. 3. Laparoscopic biliary irrigation, successful in several case studies. PMID- 9871893 TI - [Assessment of independence in daily life in pediatric oncology by FMH questionnaire]. AB - Therapy-related late effects are important for therapeutic decisions in pediatric oncology. We quantified the degree of impairment of independence in daily life in children with cancer. The German questionnaire "Fertigkeitenskala Munster/Heidelberg (FMH)" is a standardized tool for measurement of motoric and verbal functioning. A point-score leads to an age-related percentile ranking similar to typical percentiles in pediatrics. We used the FMH in 215 cancer patients (mean age 10.3 years, range 0.5-23.5 years, 56.3% male). Diagnoses were leukaemia (n = 91), bone tumors (n = 33), nephro- and neuroblastoma (n = 21), brain tumors (n = 18), lymphoma (n = 23), rhabdomyosarcoma (n = 11) and others (n = 18). The average time to answer the questionnaire was 4.5 min. Patients with brain and bone tumors showed significant lower percentile scores compared to patients with other diagnoses (p < 0.05). The FMH-scores increased with time since diagnosis (n = 215). This trend was confirmed in a longitudinal study over one year (n = 29). Quantitative assessment of independence and functioning in patients with cancer--especially in multicenter-studies--is possible. Because of therapy-related late effects this seems to be of special importance in brain and bone tumor patients. PMID- 9871894 TI - [Multiplication of chromosomes and primary leucocyte number in childhood ALL and hyperdiploid karyotype]. AB - In the blast cells of children with acute lymphoblastic leukemia (ALL) more than 50 chromosomes can be observed in a quarter of cases. As a rule these children have a good prognosis. However, some of these patients develop a relapse of their basic disease. There is only poor information about the significance of distinct additional chromosomes for the prognosis. The white blood cell count (WBC) at the time of diagnosis is a further very important prognostic factor in childhood ALL. Therefore we compared the relation between trisomy of distinct chromosomes and the initial white blood cell count of 41 children with common ALL and hyperdiploid karyotype. The modal chromosome number ranged from 50 to 60 chromosomes. Most frequently, the chromosomes X, 4, 6, 8, 10, 17, 18 and 21 were multiplied. Additionally, in 25 of the 41 cases structural chromosome aberrations were observed. The average WBC was estimated as 9.6 Gpt/l with a range from 1.8 to 41.5 Gpt/l. The initial WBC was slightly increased in patients with the additional chromosome X, 6, 11, 12 or 19 and distinctly decreased in children with the additional chromosome 8 or 9 in their hyperdiploid blast cells. No patient with an additional Chromosome 9 showed a WBC higher than 10 Gpt/l and only 2 out of the 12 children with an additional chromosome 8 revealed an initial WBC higher than 10 Gpt/l. Additional structural chromosome aberrations were without influence on the WBC. PMID- 9871895 TI - [Undescended testis and hypospadia in sex chromosomal aberrations]. AB - If hermaphrodite genitals are present in the patient or a higher degree of hypospadia is shown with maldescensus testis, a chromosomal disorder must be considered as one potential cause of the anomaly. The case report of a child with cryptorchidism on the right, inguinal testis on the left and penoscrotal hypospadia is presented as an example. A mosaic karyotype 45, X/46, X, idic (Yp) was diagnosed in this patient after chromosomal analysis. The cell line with the isodicentric Y chromosome could be demonstrated in about 90% of the lymphocytes, but only in 7% of the fibroblasts of the preputium. A derivative Y could not be detected in interphase nuclei in the buccal mucosa, i.e. only the cell line with monosomy X was presented. There was thus chromosomal mosaicism with unequal tissue involvement and a high potential for malignant transformation. Guidelines of pediatric urological, cytogenetic and endocrinological investigations and the diagnostic procedures are described and discussed. A prevention protocol for patients with comparable gonosomal mosaicism is presented. PMID- 9871896 TI - Intracerebral hemorrhage as a late complication after CNS treatment of childhood lymphoma. AB - Since the majority of children with acute leukemia and lymphoma are long-term survivors, more attention is directed towards late sequelae of therapy. Intracerebral hemorrhage after treatment of central nervous system (CNS) neoplasia in childhood is a very rare event. A seven and a half-year-old boy was admitted to our hospital because of acute third nerve palsy. Three years and eight months before the patient had been treated for a mediastinal T-cell non Hodgkin's lymphoma (T-NHL) with CNS involvement by combined chemo-radiotherapy. Recurrent disease was excluded, but intracerebral hemorrhage in the tectal area was demonstrated by repeated magnetic resonance imaging. Symptoms of incomplete oculomotor paresis improved spontaneously with conservative therapy. Intracerebral hemorrhage may occur as a rare complication in children with malignant CNS disease even years after treatment with combined chemo radiotherapy. PMID- 9871897 TI - [Immunological aspects of a child with idiopathic pulmonary hemosiderosis and celiac disease]. AB - A case report of a girl aged 3.5 years affected by the extremely rare combination of idiopathic pulmonary haemosiderosis (IPH) and coeliac disease (CD) is presented. It is the 13th such case that has been published over the last 25 years and only the 7th to be reported in a child. We believe that the concurrence of these two diseases is not coincidental, because a gluten-free diet had beneficial effects on the pulmonary symptoms not only in our case but also in other such patients. However, the pathogenetic relation between IPH and CD remains unclear. Although circulating immune complexes were detected in our patient's serum, there was no evidence of their putative damaging effect on the basement membrane of the alveolar capillaries. Furthermore, no IgA deposits could be demonstrated in alveolar basement membranes. Therefore the hypothesis that there is a reaction between IgA reticulin or endomysial antibodies and an alveolar basement membrane antigen with consecutive structural damage is unlikely. PMID- 9871898 TI - [Pigeon breeder's disease in childhood]. AB - Pigeon breeder's disease is a common form of hypersensitivity pneumonitis, which is rarely diagnosed in children. We report a seven year old boy, who developed cough, easy fatigue, anorexia and weight loss over a period of two months after having contact with pigeons. The findings on physical examination were tachypnoea and reduced thoracic movements. Chest radiograph showed widespread fine nodular shadowing in both lung fields. Pulmonary function tests demonstrated a restrictive defect and an impaired diffusion. Lymphocytes were highly increased in bronchoalveolar lavage (BAL) fluid. Precipitating antibodies against pigeon excreta was found in the serum. Environmental control of pigeon protein and a course of systemic corticosteroids over 4 months resulted in clinical and lung function improvement. However clinical recovery stopped over several months after discontinuation of steroid therapy. Therefore steroid therapy was reinstalled and continued until complete clinical recovery and pulmonary function occurred, that was 20 months after onset of the disease. Essential for the patient was to avoid further contact with birds. However a prolonged steroid therapy was necessary to achieve complete resolution. Lung function tests were useful for follow up and decisions on further treatment. PMID- 9871899 TI - [Severity of respiratory syncytial virus infection influenced by clinical risk factors and subtype A and B in hospitalized children]. AB - BACKGROUND: Since it is possible to identify the subgroups of RSV, A-subtype and B-subtype, there are findings indicating that the subtype may influence severity of RSV infection. Our study was designed to assess the hypothesis that A-subtype infections were more severe than B-subtype infections among hospitalized children. PATIENTS: All medical records of patients hospitalized with RSV infection between March 1990 and March 1993 were reviewed. A total of 107 children with proven RSV infection were identified. METHODS: Nasal waste specimens for culture were obtained from infants with suspected RSV infection. Subtype determination was done on frozen virus cultures. The following risk factors were defined: age < or = 3 months, weight < 5 kg, prematurity and underlying cardiac or respiratory disease and immune deficiency. To analyse the relationship between risk factors, subtype and severity a multivariate analysis was performed. Severity was measured by clinical observations as following: pH, PCO2, SaO2, oxygen supplementation, history of apnea and length of hospital stay. MAIN RESULTS: Of the enrolled patients 11 had underlying disease and 17 were premature. The age range was 1 week to 4.2 years, median 3.5 months. 46 children were younger than 3 months, 33 had a weight of less than 5 kg. The isolates of 84 children were typeable: 63 isolates were subtype A and 21 subtype B. Underlying disease and prematurity were associated with SaO2 < 87% (p = 0.003) and oxygen supplementation (p = 0.017). A weight of less than 5 kg was correlated with a PCO2 > or = 50 mmHg. The RSV subtype was not significantly correlated with severity. CONCLUSIONS: RSV infection even in very young children is predominantly influenced by underlying disease, prematurity and weight. The RSV subtype was no independent risk factor for an increased morbidity in this retrospective study. Therefore, in our opinion, RSV subtype is less meaningful to predict the severity of RSV infection than known risk factors. PMID- 9871900 TI - [Pneumatocele after hydrocarbon aspiration]. AB - Report of accidental ingestion and aspiration of lamp oil of a boy aged 16 months. He develops in a few hours a severe pneumonitis with tachycardia. Echocardiography shows diminished contractability. After losing his symptoms, the chest x-rays on the 13th day show bilateral pneumatoceles. The course remains asymptomatic and radiographs taken 15.5 weeks after the accident show replacement by scars. Usually the prognosis is good. Indication for invasive treatment is given when cardiorespiratory insufficiency or a rupture into the pleura space are seen. PMID- 9871901 TI - [Miliary tuberculosis and adult respiratory distress syndrome in an infant]. AB - We report on a 7 months old infant, who was admitted to our hospital with the tentative diagnosis of interstitial pneumonia. The infant was treated with erythromycin because of a positive mycoplasma antigen test in the nasopharyngeal mucus and the radiological findings. A tension pneumothorax led to rapid respiratory deterioration and a severe ARDS developed. A sustained respiratory stabilization could not be achieved. The diagnosis miliary tuberculosis was detected on autopsy. A parenchymal focus was found subpleurally in the left lung with a massive hematogenous spread and a tuberculous meningitis. The source of infection could not be identified in the family or in persons known to have had contact to the infant. The tuberculin skin test (PPD) was negative. Clinical, radiological and pathological findings are demonstrated. PMID- 9871902 TI - [Leishmaniasis. German Society for Pediatric Infectious Diseases (DGPI), Parasitology Working Group]. PMID- 9871903 TI - Ovarian ageing and postponement of childbearing. PMID- 9871904 TI - Ovarian ageing and the general biology of senescence. AB - Ovarian ageing is not only of major importance in its own right but is also of interest for its relationship with the general biology of senescence. A key feature of ageing is the distinction in higher animals between the immortality of the germ-line and the mortality of somatic cells and tissues. The ovary contains the female germ cells, and it is through these cells that the female contribution to germ-line immortality is effected. It is abundantly clear that individual oocytes can and do age and that the ageing of the ovary plays a major role in initiating or accelerating a series of other senescent changes. To understand how ovarian ageing fits within the general biology of senescence, it is necessary to explain why ageing occurs at all, to examine the likely mechanisms of general ageing, and to ask whether there is anything special about ovarian ageing and its relationship with the human menopause. Research on ovarian ageing interacts with the our emerging understanding of the general biology of senescence at many levels, ranging from the evolution of the human life history to the biochemical and cellular mechanisms of ageing and longevity. PMID- 9871905 TI - General ageing and ovarian ageing. AB - The age related decrease in female fertility is associated with a decrease in follicle numbers and oocyte quality. Meiotic division errors, mitochondrial DNA mutations and ageing itself have been suggested to play a part in the age associated reduction in oocyte quality. During the past decades several hypothesis have been proposed, trying to explain the underlying mechanisms. However, none of them is yet conclusive. This review will consider the main hypotheses regarding the age related reduction in oocyte quality. This will be reviewed together with recent results of studies analysing a possible relationship between ageing and ovarian ageing. On the basis of our own results and those presented in the literature, it is concluded that ovarian ageing may only be related to specific aspects of general ageing. PMID- 9871906 TI - Age at menopause as a marker of reproductive ageing. AB - Ages of menopause and of the preceding reproductive events such as the beginning of subfertility and infertility, are likely to be dictated by the process of follicle depletion leading to loss of oocyte quantity and quality. To some extent this process is influenced by lifestyle factors like smoking, and possibly also by the use of oral contraceptives. Genetic factors and possibly also events during intrauterine life, probably play a more important role in the age dependent decrease of female fertility. PMID- 9871907 TI - Declining fecundity and ovarian ageing in natural fertility populations. AB - Worldwide, human fertility declines with increasing maternal age, after contraceptive-use patterns and behavioral factors are taken into consideration. Here, we summarize some of our theoretical and empirical work examining the biological factors contributing to this age pattern of fertility. We undertook an 11 month prospective endocrinological study in a natural fertility (non contracepting) population (rural Bangladesh) to estimate the contributions of fetal loss and fecundability (the probability of conception) to declining fecundity with age. Prospective interviews and urine samples for pregnancy tests were collected twice weekly from up to 700 women. These data were used to test mathematical models of the underlying biological processes contributing to changing fecundability and fetal loss risk with maternal age. The results indicate that much of the decline in fecundity can be attributed to an increasing risk of fetal loss with maternal age. Much of this fetal loss is due to chromosomal abnormalities--a result of ageing oocytes. Fecundability, on the other hand, does not begin to decline until the early 40s. We hypothesize that this is also a result of ageing at the ovarian level, namely follicular atresia, in the years just prior to menopause. The irregularity of menstrual cycles- longer cycles and increasingly variable hormonal patterns--at these ages may be a direct result of the small and rapidly dwindling remaining pool of follicles. We present a simple mathematical model of this process, and some preliminary laboratory results that support the model. PMID- 9871908 TI - Ovarian follicular growth in humans: ovarian ageing and population of growing follicles. AB - By integrating morphometrical and endocrinological data, as well as biological effects of various peptides synthesized by the primate follicle, the current review proposes a morphofunctional view of the early follicle growth within the human ovary. From the time they enter the preantral stage (0.15 mm), during the early luteal phase, follicles need about 70 days to reach a size of 2 mm. These follicles are impervious to cyclic FSH and LH changes in terms of granulosa cell (GC) proliferation, and their steroidogenic activity is very low. From the time they enter the selectable stage (2-5 mm in diameter) during the late luteal phase, follicles become sensitive to cyclic changes of FSH in terms of quality and GC proliferation, but not in terms of GC aromatase activity. Thus, as the follicle develops, its responsiveness to gonadotropins progressively increases. In vitro studies suggest that this progressive acquisition of follicular cell responsiveness to gonadotropins might be under the control of peptides acting in an autocrine/paracrine fashion within the follicle. The numbers of growing follicles decrease with ageing, especially in women over the age of 40. During the follicular phase of the menstrual cycle, selectable follicles exhibit a mitotic activity of their GC higher in women older than 40 years than in younger women. PMID- 9871910 TI - Follicle culture after ovarian cryostorage. AB - OBJECTIVES: Cryostorage of ovarian cortical tissue, before devastating chemo- and/or radiotherapy for cancer, permits survival of primordial and early preantral follicles. This work aims for a system allowing the long-term maturation in vitro (IVM) of small immature oocytes up to fertilisable metaphase II oocytes. METHODS: A culture system allowing follicle attachment permitted the growth and maturation of isolated follicles (follicle diameter between 100 and 130 microns) from 14-day-old (prepuberal) mice. Follicle and oocyte development were observed under the inverted microscope and conditioned medium was used for biochemical analysis. Effects of recombinant gonadotrophins and oxygen tensions were studied for their specific effects on follicle development. RESULTS: A 12 day culture period yielded full-grown oocytes which were able to complete meiosis (metaphase II). Live young were obtained after IVF and intra-uterine transfer of in vitro matured oocytes. Growth and maturation were only successful when recombinant gonadotropins were added and when the incubator had a 20% oxygen tension. This system enabled the growth of early preantral follicles after cryopreservation: 80% of frozen and thawed follicles survived up to culture-day 12 and yielded a comparable blastocyst formation rate as in controls. CONCLUSIONS: The mouse model suggests that IVM is a valuable option after oocyte storage. The development of a comparable system for long-term culture of human follicles will imply the acquisition of non-invasive techniques to appreciate oocyte's maturity and developmental capacity. PMID- 9871911 TI - Follicle development during the normal menstrual cycle. AB - An understanding of the factors which determine initiation of follicle growth, recruitment and dominant follicle selection may increase our understanding of the underlying process of ovarian aging. In this article, these aspects of the normal menstrual cycle are reviewed. The morphological and endocrinological development in the early follicle is described from the primordial follicle stage. The degree of follicle-stimulating hormone (FSH) dependency is discussed, as is the relationship of estradiol (E2) production to follicle diameter. The principles governing mono-follicular selection are outlined, and the FSH 'threshold' and 'window' concepts are highlighted. Maximum FSH levels in the early follicular phase are shown to be variable between individuals. The relevance of this and the means by which individual sensitivity to FSH may be altered at the ovary in the context of ovarian aging are discussed. PMID- 9871909 TI - Genetics of oocyte ageing. AB - OBJECTIVES: Correlations between parental age, aneuploidy in germ cells and recent findings on aetiological factors in mammalian trisomy formation are reviewed. METHODS: Data from observations in human oocytes, molecular studies on the origin of extra chromosomes in trisomies, experiments in a mouse model system, and transgenic approaches are shown. RESULTS: Errors in chromosome segregation are most frequent in meiosis I of oogenesis in mammals and predominantly predispose specific chromosomes and susceptible chiasmate configurations to maternal age-related nondisjunction. Studies on spindle structure, cell cycle and chromosome behaviour in oocytes of the CBA/Ca mouse used as a model for the maternal age-effect suggest that hormonal homeostasis and size of the follicle pool influence the quality, maturation competence and spindle size of the mammalian oocyte. Predisposition to errors in chromosome segregation are critically dependent on altered cell cycles. Compromised protein synthesis and mitochondrial function affect maturation kinetics and spindle formation, and cause untimely segregation of chromosomes (predivision), mimicking an aged phenotype. CONCLUSIONS: Altered cell cycles and untimely resolution of chiasmata but also nondisjunction of late segregating homologues caused by asynchrony in cytoplasmic and nuclear maturation appear to be causal to errors in chromosome segregation with advanced maternal age. Oocytes appear to lack checkpoints guarding against untimely chromosome segregation. Genes and exposures affecting pool size, hormonal homeostasis and interactions between oocytes and their somatic compartment and thus quality of follicles and oocytes have the potential to critically influence chromosome distribution in female meiosis and affect fertility in humans and other mammals. PMID- 9871912 TI - Genetics, age, and infertility. AB - OBJECTIVE: To review the genetics of aging specifically as it pertains to human fertility, as well as the recent advancements in the diagnosis of genetic diseases prior to embryo implantation. METHODS: A review of our own experience as well as the scientific literature with regards to the decline in female fertility with age, the success of IVF in women of older reproductive age, and the role of preimplantation genetic diagnosis (PGD) in the evaluation of the patient at risk for fetal genetic anomalies. RESULTS: The decline in female fertility occurs primarily as a result of a decline in oocyte quality as well as quantity. The frequency of chromosomal anomalies in recognized abortuses increases in parallel with the age-specific rise in the incidence of spontaneous abortions. PGD is an accurate diagnostic tool for exclusion of genetically deficient embryos prior to initiation of pregnancy. CONCLUSION: Reproductive failure in women of older age appears to be directly related to ovarian age. Recent techniques such as cytoplasmic or germinal vesicle transfer are designed to replace the senescent cellular machinery believed to be responsible for genetic errors that occur during early cell division. PGD can accurately identify embryos with genetic deficiencies prior to implantation. PMID- 9871913 TI - Inhibin and reproductive aging in women. AB - The monotropic FSH rise is the sentinel endocrine event that first indicates a woman is approaching the end of her reproductive potential. While a deficiency in inhibin has long been postulated as the immediate cause of the monotropic FSH rise, this has only recently been demonstrated to actually occur. It is our current hypothesis that when the number of preantral follicles in both ovaries drop below a threshold, then there is a subtle decrease in inhibin B which leads to the monotropic FSH rise which, in turn, accelerates follicular depletion and the attainment of the menopause. PMID- 9871914 TI - Ovarian reserve tests in infertility practice and normal fertile women. AB - The decline in fecundity with the age of the woman is mainly attributed to the loss of follicles from the ovary and a decrease in oocyte quality. Evaluation of the aging status of the ovary in an individual woman has been hampered by a lack of knowledge with regard to the relative contribution of these two factors. Most if not all so called ovarian reserve tests (ORT) reflect indirectly the remaining follicle pool in the ovary. Direct a priori assessment of oocyte quality is not possible to date. In this section the predictive value of several ovarian reserve tests for the outcome of fertility treatment is listed and commented. In addition, the study of several of the ORTs in normal, fertile women is described. From the data presented dynamic testing of the ovarian function by the clomiphene citrate and GnRH agonist stimulation test, as well as static testing by the use of ultrasound based antral follicle counts seem to offer the highest clinical value. Studies performing direct comparison of these tests are needed, as well as analysis of the way these tests should direct decision making in infertility diagnosis and treatment. PMID- 9871915 TI - Interpretation of elevated FSH in the regular menstrual cycle. AB - In reproductive medicine, abnormal elevation of serum follicle stimulating hormone (FSH) concentrations during the luteofollicular transition is associated with low response in infertility treatment. Increasing levels of serum FSH in the early follicular phase is a characteristic of reproductive ageing and has become very popular for the determination of diminished ovarian oocyte reserve. In the case of elevated FSH in premenopausal women, many more (patho)physiological mechanisms other than ageing may be responsible and should be considered also. FSH concentrations may vary considerably due to a number of factors. Next to intra-, inter- and between different assays variation there is hourly-, cycle day dependent-, intercycle and life time variation. Furthermore, physiological conditions such as during puberty, in hereditary dizygotic twinning, after use of oral contraceptives and during lactation also elevated FSH levels can be found. Pathological conditions associated with significant increases in FSH are after unilateral ovariectomy, during recovery from hypothalamic amenorrhea and with excessive smoking. It should be kept in mind, that with the interpretation of an abnormal high FSH value, just as with any other abnormal laboratory results, one should be aware, that there may be a variety of underlying causes other than the one where was aimed at. PMID- 9871916 TI - The impact of age on reproductive potential: lessons learned from oocyte donation. AB - OBJECTIVE: Oocyte donation allows a unique opportunity to separately study the effect of aging on uterine receptivity and oocyte quality. The purpose of this report is to review the published experience on reproductive aging in both laboratory animals and humans as it pertains to oocyte and embryo donation. METHODS: A review of the published medical literature. RESULTS: Natural fertility rates decline in most animals with age, becoming dramatically apparent in women as they enter the fifth decade of life. By the time of the perimenopause, pregnancy rarely occurs, whether or not assisted reproductive techniques are initiated. However, if oocytes are donated by young women to older women, both embryo implantation and pregnancy rates are restored to normal levels in recipients. CONCLUSIONS: These results strongly suggest the pregnancy wastage experienced by older women is largely a result of degenerative changes within the aging oocyte, rather than senescent changes in the uterus. The poor prognosis for fertility in older women can be reversed through oocyte donation from younger individuals. PMID- 9871917 TI - Cardiac contractility: how calcium activates the myofilaments. AB - In both cardiac and skeletal muscle, the force-generating molecular motors (crossbridges) are turned on by increasing the intracellular free calcium level that regulates the troponin-tropomyosin system. However, calcium activation is a two-way process in the sense that activated crossbridges also affect the troponin tropomyosin system. Here we review the mechanism of calcium action on myofilament proteins, particularly tropomyosin, that affects both the extent and the rate of force development and hence the contractility of the myocardium. At low myoplasmic Ca2+ concentrations tropomyosin is located at the edge of the thin filament, thereby interfering with the formation of strong actin-myosin linkages (blocked state). An increase in Ca2+ activity causes an azimuthal shift of tropomyosin around the filament (by about 30 degrees), thereby increasing the probability of low-force crossbridge interaction, a process which by cooperative effects induces further tropomyosin movement (by an additional 10 degrees) which results in the open state of the filament characterized by forceful crossbridge interaction. (This mechanism may be analogous to that in ligand-gated ion channels, where ligand binding increases the open probability of the pore.) The extent of activation then depends on the free Ca2+ concentration and on the calcium sensitivity of the thin filament that may be affected by protein phosphorylation, crossbridge attachment, the troponin isoform composition of the filament, and novel calcium-sensitizing drugs that act on the contractile or regulatory proteins and thus increase the force of the heart. PMID- 9871918 TI - Coupling physiology and gene regulation in bacteria: the phosphotransferase sugar uptake system delivers the signals. AB - In many bacteria a crucial link between metabolism and regulation of catabolic genes is based on the phosphotransferase sugar uptake system (PTS). We summarize the mechanisms of the signaling pathways originating from PTS and leading to regulation of transcription. A protein domain, called PTS regulation domain (PRD), is linked to many antiterminators and transcriptional activators and regulates their activity depending on its state of phosphorylation. Two sites can be phosphorylated in most PRDs: HPr-dependent modification at one site leads to activation while enzyme II dependent phosphorylation of the other site renders it inactive. In addition, PTS components are used to generate cofactors for regulators of transcription. The paradigm is the enzyme II dependent activity of adenylate cyclase determining the cyclic AMP level in Escherichia coli and thereby the activity of the catabolite activator protein. In many gram-positive bacteria catabolite repression is mediated by the catabolite control protein CcpA, which requires HPr Ser-46 phosphate as a cofactor to regulate transcription of catabolic genes. HPr Ser-46 phosphate is produced by HPr kinase, the activity of which is under metabolic control via the concentrations of glycolytic intermediates. These recent results establish a multifaceted regulatory role for PTS in addition to its well-established function in active sugar uptake. PMID- 9871919 TI - A biomimetic electron releasing device: capacitively connected glow discharge electrode. PMID- 9871920 TI - [Labrum lesions in residual dysplasia of the hip joint. Biomechanical considerations on pathogenesis and treatment]. AB - The capsular-labrum-complex consists of the triangular fibrocartilaginous labrum, which is fixed at the bony acetabular ring and supported by the ligamentum transversum actetabuli in the caudal part. In a normally developed and correctly orientated acetabulum, the femoral head is symmetrically covered by the contact area of the lunate surface. The vertical component of the resultant hip joint force concentrically meets the horizontal weight bearing surface: Evenly distributed compressive forces are transmitted; the capsular-labrum-complex does not have to compensate excessive shear or tension. In contrast, in a "dysplastic" acetabulum the femoral head is poorly covered by the pathologically orientated lunate surface: The lunate surface is sloping in an anterolateral direction, leading to tension and shear on the superolateral capsular-labrum-complex, which becomes a secondary stabilizer ("guide rail") against the decentering femoral head. The vertical component of the resultant hip force eccentrically meets the oblique weight bearing surface, causing extra stress in the labrum: The capsular labrum-complex is trying to compensate this biomechanical stress with hypertrophy (Type IB). Progressive mechanical decompensation of the capsular-labral-complex might lead to tears (Type IIB) or complete avulsions (Type IIIB) of the labrum. In conclusion labral lesions result from pathobiomechanics caused by residual hip dysplasia (RHD). Logically, the basic therapeutic principle can only be corrective osteotomy. PMID- 9871921 TI - [Acetabular rotations y triple pelvic osteotomy by the Tonnis method]. AB - Lesions of the acetabular labrum should be treated by correcting the causes. In a steep acetabulum where the femoral head brings the acetabular labrum under tension and traction, the acetabulum should be rotated by triple pelvic osteotomy to slightly over-corrected acetabular measurements. Severely diminished acetabular and femoral anteversion can also lead to tears and impingement of the labrum. Then rotation of the femoral neck and/or rotations of the acetabulum by triple osteotomy to 15-20 degrees of anteversion are indicated. Our triple osteotomy technique differs from that of others mainly in the ischial osteotomy. It is performed from the posterior approach between the sciatic notch next to the ischial spine and the obturator foramen and is directed 20-30 degrees anteriorly from the frontal (coronal) plane. The osteotomies therefore are placed close enough to the acetabulum to allow free rotation, but they do not interfere with the circulation of the acetabulum, and the ligaments between the sacrum and ischium are left in normal tension. Our normal values of the acetabular position were tested by correlating the measurements with the absence of pain. The optimum is reached with a CE angle and a VCA angle of 30-35 degrees, an angle of the weight-bearing zone of +5 to -5 degrees and a migration index of 10-15%. Overcorrections again caused pain and should be avoided. Diminished anteversion of femur and acetabulum towards 0 degree also caused pain and should be corrected by triple and femoral osteotomy to 15-20 degrees of anteversion. In earlier follow-ups of 216 hips 5-10 years postoperatively, 82.3% of the joints showed no change in the degree of osteoarthrosis. Survival rate curves regarding the absence of pain demonstrated that pain was experienced again when joints were corrected insufficiently or overcorrected, while in good corrections the joints were free of pain in about 75%. PMID- 9871922 TI - [The Bernese method of periacetabular osteotomy]. AB - The periacetabular osteotomy is a strategy for the treatment of residual hip dysplasia in young adults with the aim of preventing secondary coxarthrosis. This polygonal, juxta-articular osteotomy respects the vascular blood supply to the acetabular fragment and thus facilitates an extensive acetabular reorientation with improvement of the insufficient coverage of the femoral head, including medial displacement of the fragment. All osteotomies are performed by one approach, the modified Smith-Petersen, which allows an anterior capsulotomy. Inspection of the joint not only provides information on acetabular rim pathologies, but also facilitates the control of impingement-free range of motion after the correction. The posterior column remains partially intact, allowing minimal internal fixation of the acetabular fragment and early mobilization similar to that after an intertrochanteric osteotomy. The dimensions of the true pelvis are unchanged, providing the capacity to have an unimpeded delivery in women. This paper describes the preoperative evaluation, current indications, surgical technique, postoperative care, and the results of this osteotomy. PMID- 9871923 TI - [The Boston concept. peri-acetabular osteotomy with simultaneous arthrotomy via direct anterior approach]. AB - To this point, we have only a relatively short term followup in 32 patients with known labral lesions who have been treated by combined periacetabular osteotomy (PAO) and debridement of the damaged anterior labrum. A positive clinical history in association with pain demonstrated with the impingement test of flexion adduction-internal rotation nearly always indicates macroscopic pathology within the anterior portion of the hip joint. Labral lesions associated with acetabular dysplasia seem more common with increasing age. Labral lesions seem to correlate with arthrosis. We suspect that uncorrected impinging lesions of the anterior rim have led to some of our early clinical failures after PAO. We feel that intra articular treatment of certain impinging lesions may improve the clinical outcome in these patients. Correction of the pathological mechanics in the mature, dysplastic hip certainly requires restoration of stability by reorienting osteotomy. If intra-articular derangements of the anterior rim are also present, though, intra-articular surgery may also be necessary to optimize the outcome. Our early results suggest that the earlier and more complete the correction of the disordered mechanics in the dysplastic hip, the more complete and long lasting will be both relief of clinical symptoms and preservation of a joint free from arthrosis. Anterior arthrotomy to explore the anterior rim, carried out at the time of PAO, employing the direct anterior abductor-sparing approach (DAA), seems a safe and useful adjunct in treating the mature, dysplastic hip. Much longer clinical followup and larger treatment groups will be necessary to allow firm conclusions concerning optimal treatment programs for different patient subgroups of the adult hip dysplasia syndrome. PMID- 9871924 TI - [The Osaka concept. "Dome osteotomy" with of without labrum resection]. AB - Acetabular dysplasia represents the most common aetiology for secondary osteoarthritis of the hip joint in Japan. On radiographs progress of the disease can be classified in four stages (I to IV). Labral lesions are common accompanying findings in acetabular dysplasia, representing an important prognostic factor. Therefore we routinely use conventional arthrography to classify labral lesions in three types (normal, torn and detached). From 1978 to 1983 a modified Chiari osteotomy of the pelvis (dome osteotomy) was performed in 64 hip joints without arthrotomy. The preoperative radiography showed labral lesions in 66%. After an average follow-up of 4 years, 43 of the 44 patients with a normal or torn labrum showed excellent or good results. On contrast, 50% of the 20 patients with a detached labrum showed fair and poor results only. From 1984 to 1989 an arthrotomy was combined with dome osteotomy in 29 hip joints and labral lesions had to be resected in 16 cases. The results were generally good in patients with arthrosis grade I and II, but in stage III they were only fair. Thus, the success of labral surgery can only be achieved in early stages of dysplasia. Based on our findings dome osteotomy combined with arthrotomy and labral surgery can be recommended in patients with labral lesions and hip dysplasia grade I and II. PMID- 9871925 TI - [The Donauspital-Stolzalpe concept (Danube Hospital, Vienna, Austria). Corrective osteotomy with selective labrum surgery following preoperative MRI arthrography]. AB - Concomitant pathologies (labral lesions, intra-/extra-osseous ganglia and stress bone marrow edema) in adult residual hip dysplasia (RHD) might influence the outcome of conservative hip surgery. The aim of our prospective clinical study was to assess the value of preoperative MR arthrography in diagnosing concomitant lesions and in making surgical decisions in RHD. The first 37 consecutive patients with a minimum follow-up of 18 months have been analysed. All 37 patients presented RHD with the clinical symptomatology of labral lesions and underwent routine preoperative MRA. According to clinical, radiological and MR arthrographical criteria, these 37 patients were subdivided into four therapeutic subgroups: (1) reorientation of the acetabulum using the Tonnis triple pelvic osteotomy (TPO); (2) intertrochanteric varisation osteotomy (IVO); (3) palliative decompression with only symptomatic partial resection of the torn labrum (PALL); (4) primary total hip replacement (TEP). Based on the preliminary clinical and radiological outcomes of these four subgroups, the following conclusions can be drawn: labral lesions are considered to be a sign of chronic joint instability. Therefore, acetabular malorientation should be corrected by redirectional osteotomy of the acetabulum (TPO-subgroup) even in low grades of RHD if labral lesions are present. "Palliative" labral resections without corrective osteotomy (PALL subgroup) in secondary osteoarthritis due to RHD are definitively obsolete, because they rapidly progress to severe osteoarthritis due to surgically accelerated joint instability. In RHD with highly osteoarthritic hip joints and concomitant lesions, one should not hesitate to perform primary THR even in young patients. PMID- 9871926 TI - [Labrum lesions and residual dysplasia of the hip joint. Definition and prospectives]. AB - Labral lesions are a sign of biomechanical decompensation of the hip joint and often represent the first clinical symptom of residual hip dysplasia (RHD) in the adult. Provocation tests (impingement, apprehension) are typical but not specific. Labral lesions and concomitant findings (intra- and extraosseous ganglia, stress bone marrow oedema) can be detected by magnetic resonance arthrography (MRA) with an accuracy of 91%. Primary therapeutic goal is the normalization of the underlying pathomorphology and instability by a redirectional acetabular osteotomy. There are several concepts concerning simultaneous arthrotomy at the time of osteotomy: no arthrotomy at all, selective arthrotomy, routine arthrotomy in every case. There are more clinical studies necessary before one of these concepts can be widely accepted and recommended. Based on preliminary results, palliative arthrotomy with partial labral resection but without corrective osteotomy in osteoarthritis secondary to residual hip dysplasia gives poor results; we therefore urgently dissuade from palliative labral surgery via arthrotomy. Whether labral surgery via arthroscopy might be a useful concept in symptomatic residual hip dysplasia, is still an open question. In this review article, the "state of the art" presented at the "Vienna Labral Symposium 1997" is reflected and summarized. At the end of this article, a "common statement" of the experts is published in English and German language. PMID- 9871927 TI - [Amputation and prosthetics. 2: Amputation of the foot and its orthopedic management]. PMID- 9871928 TI - [Interleukin 10: a key cytokine in disseminated lupus erythematosus]. PMID- 9871929 TI - [Growing incidence of nalidixic acid resistance and sensitivity to quinolones in Salmonella typhimurium strains isolated from man or animal]. AB - To determine the prevalence of quinolone resistance in Salmonella typhimurium strains from humans or animals (cattle, poultry, swine), the S. typhimurium strains isolated at a teaching hospital and at the central veterinary laboratory of the same district between January 1, 1995, and December 31, 1996 were studied. Susceptibility to nalidixic acid was determined using the disk diffusion method. Strains with decreased susceptibility to nalidixic acid were subjected to minimal inhibitory concentration (MIC) determination for pefloxacin, ofloxacin, ciprofloxacin, norfloxacin, levofloxacin, and grepafloxacin. Decreased susceptibility to nalidixic acid was demonstrated for 41 of the 309 strains studied and increased from 8.5% in 1995 to 18.6% in 1996. MIC90 values of fluoroquinolones for strains with decreased susceptibility to nalidixic acid were lower than 1 mg/L, which is the cutoff above which a strain is classified as susceptible, but were higher than for strains that were susceptible to nalidixic acid. These low levels of resistance may be the first step in selection of mutant strains with high levels of resistance to fluoroquinolones. This warrants continued monitoring of resistance of Salmonella to fluoroquinolones. PMID- 9871930 TI - Validation of cardiac echocardiography for measuring cardiac output to be applied for the multiple inert gas elimination technique. AB - The multiple inert gas elimination technique (MIGET) is being increasingly used in respiratory physiology and pathophysiology. Six inert gases are given as an intravenous infusion then measured in samples of expired air and mixed arterial and venous blood. This requires right-sided catheterization, a procedure that is sometimes ethically inappropriate. The present article reports a method in which inert gas levels in mixed venous blood were calculated, rather than measured, using Fick's law. Echocardiography was used to measure arterial inert gas levels and cardiac output. The method was validated in 11 men scheduled to undergo coronary bypass surgery. Cardiac output was either calculated based on biometrical (C) data or measured using four different methods in random order, namely Fick's law with oxygen (FiO2) or the inert gases (FiIG) as the tracers, thermodilution (TH), and echocardiography (E). Cardiac output values in L.min-1 (mean +/- SD) were as follows: C, 4.99 +/- 0.39; FiO2, 5.44 +/- 0.86; FiIG, 5.55 +/- 0.92; TH, 5.77 +/- 0.88; and E, 5.53 +/- 0.64. No significant differences were found among the four measured cardiac output values, of which the mean was 5.57 +/- 0.70 L/min, a value that was significantly higher than the calculated value. This difference is probably ascribable to the use of dopamine, dobutamine, or epinephrine in six of the 11 patients. A 1 L/min-1 cardiac output error, in either direction, was found to have a marked influence on the distribution of alveolar perfusion at various VA/Q ratios. Conversely, as expected, ventilation distribution was not influenced by cardiac output. In conclusion, echocardiography provides satisfactory cardiac output estimations using the MIGET except in patients with septal hypertrophy, subaortic membranes, a mitral valve prosthesis, or a mitral valve ring. PMID- 9871931 TI - Diagnosis of extrapulmonary tuberculosis by a commercial polymerase chain reaction kit. AB - The Roche Amplicor Mycobacterium tuberculosis PCR test was compared with mycobacterial culture for direct detection of M. tuberculosis in extrapulmonary specimens. From January 1995 to October 1996, 124 clinical specimens from 112 patients were assessed, including 47 body fluids, 61 tissue specimens and 16 abscesses. The sensitivity and specificity of Amplicor PCR compared to culture were 63.6% and 93.1% respectively. Analysis of 7 PCR-positive, culture-negative specimens confirmed that all were from patients with recently diagnosed tuberculosis under treatment. Eight specimens were PCR negative-culture positive, including a pleural fluid containing inhibitory substances. On acid-fast bacilli (AFB) smear-negative specimens, sensitivity and specificity were 53 and 100% respectively. The best results for Amplicor PCR were obtained with abscesses and biopsies. It is concluded that this test, highly specific for the diagnosis of tuberculosis, is at least as sensitive on extrapulmonary specimens as on smear negative respiratory specimens. PMID- 9871932 TI - [Hemoglobin alpha chain isoelectric point modification under the action of urea, sodium cyanate, succinic anhydride or diethylene triamine pentaacetic acid anhydride]. AB - A monomeric protein, the hemoglobin alpha chain, was used to compare four protocols for conjugation with diethylene triamine pentaacetic (DTPA) anhydride. Carbamylation and succinylation were also performed. The isoelectric point (pI) was 7.7 for the native protein versus only 5.5 to 7.3 for the five carbamylated derivatives and 4.0 to 7.0 for the six succinylated derivatives. With carbamylation or succinylation, increasing the molar ratio (agent/protein) was associated with a gradual downward pI shift producing trains of bands. This phenomenon did not occur with DTPA conjugation, whose results varied with the method used; only one derivate (pI 6.7) was produced by all four methods, and multiple fine bands with pH values in the vicinity of 3.6 were seen. For the protein, the pI shift varied with the number of groups inserted on the primary amine residues. Also, the shift was larger if the inserted groups carried electrically-charged moieties. PMID- 9871933 TI - [Evaluation of the Widal test for diagnosing typhoid fever in Lebanon]. AB - Typhoid fever is a major public health problem in developing countries. In Lebanon, where the sanitary infrastructure was severely damaged during the civil war, the high prevalence of typhoid fever creates a pressing need for a rapid and reliable diagnostic tool. To evaluate the diagnostic usefulness of the Widal test, patients with a positive blood culture for Salmonella typhi (n = 78) were compared to controls with fever due to another infection (n = 65) and with blood donors (n = 96). Discrimination between the patient group and the two control groups was best when an agglutinin O titer of 1/160 or more was used. With this cutoff, sensitivity was 67.9%; specificity was 93.8% versus the febrile controls and 100% versus the blood donors. These results suggest that despite its well known limitations the Widal test remains a valuable tool for typhoid fever control in many developing countries. PMID- 9871934 TI - [Granzyme B: an essential protease for the inflammatory response]. AB - Granzyme B is a serine protease produced by cytotoxic lymphocytes and capable of inducing rapid target cell death by apoptosis. This effect was found to be closely correlated with the presence of perforin, or pore-forming protein, also contained in the cytoplasmic granules of cytotoxic lymphocytes. Subsequent data suggested that the chief role of perforin is to facilitate accessibility of granzyme B to its cytoplasmic and nuclear targets. Granzyme B may penetrate within the cytoplasm autonomously via a membrane receptor expressed by the target cell, before entering endocytic vesicles containing the protein. Granzyme B can induce target cell death via two complementary pathways, a cytosolic pathway involving cascade activation of proapoptotic caspases, and a nuclear pathway probably involving a cell cycle regulating protein and/or kinase Cdc2 activation. Recent data have established that non-lymphoid cells, including human epithelial cells, can express granzyme B and release it into the extracellular space during the repair process following disruption of the epidermal barrier. This results in local anti-infectious activity that compensates for the break in the mechanical skin barrier and may be a component of natural immunity. PMID- 9871935 TI - [Molecular typing of mycobacteria]. AB - Molecular typing is now widely used in mycobacteriology as a significant addition to conventional epidemiological studies. Several methods are available that differ with respect to their discriminating power and to the genetic marker used. Restriction fragment length polymorphism (RFLP) done using the IS6110 insertion sequence has been widely used for typing Mycobacterium tuberculosis. More recently, short repetitive DNA sequences have been used as RFLP markers, as well as for other typing methods involving gene amplification. One of these methods, the inter-direct repeat (DR)-based spoligotyping method, is rapid, directly applicable to clinical specimens, and highly discriminating. PMID- 9871936 TI - [Molecular methods for the rapid diagnosis of mycobacterial infections]. AB - With the aim to reduce the time for the diagnosis of mycobacterial infections, various molecular methods were developed recently which are particularly suitable for clinical mycobacteriology laboratories. These methods permit both the detection and rapid identification either directly from the pathological sample or culture. We hereby review the most recent advances in this field, particularly those involving hybridization with molecular probes, or nucleic amplification methods. We also review the strategy employed for current research developments for application in mycobacterial diagnosis as well as the future prospects. PMID- 9871937 TI - [Comparative activity of cefepime, ceftazidime and imipenem in a mouse infection caused by Klebsiella pneumoniae producing a broad spectrum beta-lactamase]. AB - Mice experimentally infected with extended-spectrum beta-lactamase-producing Klebsiella pneumoniae strains were injected twice daily for three days with ceftazidime, cefepime, or imipenem (25, 50, or 100 mg/kg/injection). Treatment efficacy was based on five-day survival and on the spleen viable bacteria count 16 hours after the last treatment dose. Under these experimental conditions, ceftazidime showed some activity on strains with low levels of resistance to ceftazidime. Cefepime used in a dose of 50 or 100 mg per injection demonstrated good activity but was slightly less effective than imipenem. PMID- 9871938 TI - [Antibacterial activity of ofloxacin in urine for 4 days after a single oral dose of 400 mg]. AB - Antibacterial activity of ofloxacin in urine after a single oral dose of 400 mg was evaluated in ten healthy female volunteers. Urine was collected over six periods, i.e., 0-6 h, 6-12 h, 12-24 h, 24-48 h, 48-72 h, and 72-96 h postdose. Ofloxacin levels were assayed in all samples using a microbiological method and HPLC. Urinary ofloxacin MICs were determined for five bacterial strains recovered from urine, two E. coli strains of which one was susceptible and the other resistant to nalidixic acid (Nal-A), one Klebsiella pneumoniae resistant to nalidixic acid (Nal-B), one Staphylococcus saprophyticus strain, and one Enterococcus faecalis strain; MICs were 0.06, 0.25, 1, 0.25, and 2 mg/L, respectively. Mean urinary ofloxacin levels by the microbiological method during the six collection periods were 193.3 +/- 30.3, 138.1 +/- 31, 53.2 +/- 7.3, 8.3 +/- 0.8, 1.4 +/- 0.2, and 0.6 +/- 0.1 mg/L, respectively. HPLC provided similar results: 216.7 +/- 31.6, 130.7 +/- 20.5, 56.5 +/- 7.1, 8.3 +/- 0.9, 1.5 +/- 0.3, and 0.5 +/- 0.05 mg/L, respectively. Mean urinary ofloxacin excretion over 96 h was 67.4 +/- 3.6% of the dose by the microbiological method was 72.5 +/- 2.5% of the dose by HPLC. On the first day, bacteriostatic activity of urine against enterobacteria exceeded 32 and was greater than 8192 for the nalidixic acid susceptible E. coli strain; on the next day, overall values were equal or greater than 8 for the nalidixic acid-resistant E. coli and K. pneumoniae strains. Bacteriostatic activity was equal to or greater than 32 for the S. saprophyticus strain during the first two days and equal to 8 on the first day and 4 on the second day for the E. faecalis strain. PMID- 9871939 TI - Tremulous jaw movements in rats: a model of parkinsonian tremor. AB - Several pharmacological and neurochemical conditions in rats induce 'vacuous' or 'tremulous' jaw movements. Although the clinical significance of these movements has been a subject of some debate, considerable evidence indicates that the non directed, chewing-like movements induced by cholinomimetics, dopamine antagonists and dopamine depletions have many of the characteristics of parkinsonian tremor. These movements occur within the 3-7 Hz peak frequency range that is characteristic of parkinsonian tremor. Tremulous jaw movements are induced by many of the conditions that are associated with parkinsonism, and suppressed by several different antiparkinsonian drugs, including scopolamine, benztropine, L DOPA, apomorphine, bromocriptine, amantadine and clozapine. Striatal cholinergic and dopaminergic mechanisms are involved in the generation of tremulous jaw movements, and substantia nigra pars reticulata appears to be a major basal ganglia output region through which the jaw movements are regulated. Future research on the neurochemical and anatomical characteristics of tremulous jaw movements could yield important insights into the brain mechanisms that generate tremulous movements. PMID- 9871940 TI - Measuring reward with the conditioned place preference paradigm: a comprehensive review of drug effects, recent progress and new issues. AB - This review gives an overview of recent findings and developments in research on brain mechanisms of reward and reinforcement from studies using the place preference conditioning paradigm, with emphasis on those studies that have been published within the last decade. Methodological issues of the paradigm (such as design of the conditioning apparatus, biased vs unbiased conditioning, state dependency effects) are discussed. Results from studies using systemic and local (intracranial) drug administration, natural reinforcers, and non-drug treatments and from studies examining the effects of lesions are presented. Papers reporting on conditioned place aversion (CPA) experiments are also included. A special emphasis is put on the issue of tolerance and sensitization to the rewarding properties of drugs. Transmitter systems that have been investigated with respect to their involvement in brain reward mechanisms include dopamine, opioids, acetylcholine, GABA, serotonin, glutamate, substance P, and cholecystokinin, the motivational significance of which has been examined either directly, by using respective agonist or antagonist drugs, or indirectly, by studying the effects of these drugs on the reward induced by other drugs. For a number of these transmitters, detailed studies have been conducted to delineate the receptor subtype(s) responsible for the mediation of the observed drug effects, particularly in the case of dopamine, the opioids, serotonin and glutamate. Brain sites that have been implicated in the mediation of drug-induced place conditioning include the 'traditional' brain reward sites, ventral tegmental area and nucleus accumbens, but the medial prefrontal cortex, ventral pallidum, amygdala and the pedunculopontine tegmental nucleus have also been shown to play important roles in the mediation of place conditioning induced by drugs or natural reinforcers. Thus, although the paradigm has also been criticized because of some inherent methodological problems, it is clear that during the past decade place preference conditioning has become a valuable and firmly established and very widely used tool in behavioural pharmacology and addiction research. PMID- 9871941 TI - REM sleep and nefazodone. PMID- 9871942 TI - Characteristics of apneas and hypopneas during sleep and relation to excessive daytime sleepiness. AB - STUDY OBJECTIVES: One of the most important symptoms in patients evaluated for possible obstructive sleep apnea syndrome is excessive daytime sleepiness, but the measures of apnea severity and of sleepiness used most commonly have not generally shown strong associations. We explored whether information recorded during standard polysomnography, other than the overall rate of apneas and hypopneas per hour of sleep (AHI), might help explain the measured severity of sleepiness. DESIGN: Observational SETTING: A clinical sleep laboratory in a university hospital PATIENTS: N = 1,146 patients evaluated for suspected sleep disordered breathing with nocturnal polysomnograms and multiple sleep latency tests. RESULTS: The AHI during supine sleep (recorded in a subgroup of n = 169 subjects), the rate of apneas (n = 1,146), and the rate of obstructive apneas (n = 1,146) were particularly useful in explaining variation in measured levels sleepiness; rates of hypopneas and central apneas were less useful (n = 1,146). In addition, the minimum recorded oxygen saturation (n = 1,097) was as important as the AHI to the level of sleepiness. CONCLUSIONS: In an attempt to explain excessive daytime sleepiness among patients evaluated for sleep-disordered breathing, additional insight is provided by observation of supine sleep during polysomnography, by emphasis on apneas rather than hypopneas, by emphasis on obstructive rather than central events, and by consideration of the minimum oxygen saturation. PMID- 9871943 TI - Obstructive sleep apnea and acute myocardial infarction: clinical implications of the association. AB - It is not known whether patients with acute myocardial infarction (MI) with coexistent obstructive sleep apnea (OSA) have a poor clinical course during the acute phase of MI. Therefore, we investigated the impact of OSA on in-hospital morbidity and mortality during an acute MI. Patients admitted to the intensive cardiac unit (ICU) with acute MI underwent Holter monitoring and night pulse oximetry (SpO2). During the first complete day at the ICU, both recordings (ECG and SpO2) were matched in time to determine association between cardiac arrhythmias and hypoxemia episodes. We identified and compared 55 heavy snorers with daytime sleepiness who showed more than 10 episodes of desaturation per hour on pulse oximetry (OSA group), and 196 nonOSA patients. There was an increase in the incidence of premature ventricular contraction (PVC, p < 0.05) and couplets PVC (p < 0.05) in OSA patients; the proportion of those arrhythmias increased in parallel with desaturation episodes in the OSA group. There were no differences between OSA and nonOSA groups for major MI complications (38.2% vs 34.2%, p > 0.05), ICU/hospital stay (3.6 +/- 1.2 vs 2.7 +/- 0.9 days, p > 0.05), or mortality within 30 days (14.5% vs 12.2%, p > 0.05). In conclusion, despite the greater incidence of some types of cardiac arrhythmias during an acute MI in OSA, these patients have the same clinical course in hospital and mortality rate as nonOSA patients. PMID- 9871944 TI - Fuzzy assessment of sleep-disordered breathing during continuous positive airway pressure therapy. AB - We propose a new method of quantifying sleep-disordered breathing (SDB) for the purpose of automating continuous positive airway pressure (CPAP) titration. Our algorithm, based on fuzzy logic, emulates the less-than-crisp kind of decision making generally employed at the human level. Three input variables were first derived on a breath-by-breath basis from respiratory airflow measurements. These were: (1) the relative duration of inspiratory flow limitation in each breath; (2) the degree of hypopnea relative to the past 15 breaths; and (3) the intensity of snoring. Using these descriptors as inputs, our fuzzy inference algorithm produced a "severity index" (SI) quantifying the degree of SDB. Severity index was determined in CPAP titration procedures conducted on one normal snorer and 12 patients with moderate-to-severe obstructive sleep apnea. SI computed over the last 6 minutes of each CPAP level was compared against other more-conventional indices of SDB, such as total pulmonary resistance (RL), the number of apneas and hypopneas (NAH), and the number of arousals (NAr). In all but one of the subjects, the correlation coefficients for SI vs each of RL, NAH, and NAr were significantly different from zero, but not different from each other. The group correlation coefficients for SI vs RL, NAH, and NAr were 0.89, 0.86, and 0.87, respectively, demonstrating that SI accurately quantifies SDB. SI collapses multiple features of the airflow pattern into a single index and, therefore, may be useful as a "feedback" variable for the automatic control of CPAP therapy. PMID- 9871945 TI - Treatment of obstructive sleep apnea syndrome by rapid maxillary expansion. AB - The precise role of maxillary constriction in the pathophysiology of obstructive sleep apnea (OSA) is unclear. However, it is known that subjects with maxillary constriction have increased nasal resistance and resultant mouth-breathing, features typically seen in OSA patients. Maxillary constriction is also associated with alterations in tongue posture which could result in retroglossal airway narrowing, another feature of OSA. Rapid maxillary expansion (RME) is an orthodontic treatment for maxillary constriction which increases the width of the maxilla and reduces nasal resistance. The aim of this pilot study was to investigate the effect of rapid maxillary expansion in OSA. We studied 10 young adults (8 male, 2 female, mean age 27 +/- 2 [sem] years) with mild to moderate OSA (apnea/hypopnea index-AHI 19 +/- 4 and minimum SaO2 89 +/- 1%), and evidence of maxillary constriction on orthodontic evaluation. All patients underwent treatment with RME, six cases requiring elective surgical assistance. Polysomnography was repeated at the completion of treatment. Nine of the 10 patients reported improvements in snoring and hypersomnolence. There was a significant reduction in AHI (19 +/- 4 vs 7 +/- 4, p < 0.05) in the entire group. In seven patients, the AHI returned to normal (i.e., = < 5); only one patient showed no improvement. These preliminary data suggest that RME may be a useful treatment alternative for selected patients with OSA. PMID- 9871946 TI - REM sleep deprivation in monocularly occluded kittens reduces the size of cells in LGN monocular segment. AB - STUDY OBJECTIVES: In this study, we test the hypothesis that when REM-state activation (which impinges upon all lateral geniculate nucleus laminae irrespective of stimulating eye) is deprived, the monocular segment (MS) that is cut off from visual input and also deprived of REM-state activation will exhibit smaller cells, owing to the loss of extrinsic as well as intrinsic activation. DESIGN: We carried out a study comparing soma sizes in the MSs of kittens subjected to monocular deprivation (MD) + REM deprivation (RD) to two age-matched nonRD groups, MD ONLYs and MD MOMS (MD kittens living in their home cages). MEASUREMENTS AND RESULTS: Perikaryal outlines of 100 cells in each of the bilateral MSs were measured. As predicted, mean cell size in the MS connected to the patched eye of MD + RD kittens, but in neither of the control groups, was significantly smaller than in the MS afferented by the nonpatched eye. One-way ANOVAs comparing MS cell-size means from the same sides across groups were also significant, but the two MSs showed different results on post hoc tests. The ordering of MS cell-size means correlated significantly with a measure that aggregates the sources of activation reaching a particular MS and their durations. CONCLUSIONS: These results reveal that removal of REM-state activation during CNS development amplifies the plasticity processes generated when normal visual afferentation to central visual areas is interrupted. Our findings in the MS of the LGN indicate that during the usual operation of REM sleep, central visual-sensory sites receive intrinsic activation that, in the visual system, is additive and complementary to the stimulation obtained from extrinsic sources. In the course of early development, normative symmetrical activation of central visual areas during REM sleep may counterbalance plasticity changes caused either by absent or aberrant sensory stimulation. PMID- 9871947 TI - Sleep architecture in a canine model of obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) causes recurrent sleep disruption that is thought to contribute to excessive daytime sleepiness in patients with this disorder. The purpose of this study was to determine the specific effects of OSA on overall sleep architecture in a canine model of OSA. The advantage of this model is that sleep during long-term OSA can be compared to both normal sleep before OSA and recovery sleep after OSA. Studies were performed in four dogs in which sleep-wake state was monitored continuously by a computer that received telemetered EEG and EMG signals. Whenever sleep was detected, the computer sent a signal to close a valve through which the dog breathed; when the dog awoke the occlusion was released. In each dog, data were analyzed from 4 consecutive nights in three phases: a control phase before induction of OSA, a phase during long-term OSA (mean = 85 days, apnea index = 59/hour), and a recovery phase after cessation of OSA. During recovery there was a significant increase in the amount of rapid-eye movement (REM) sleep compared to the OSA phase (p < 0.01), as well as significant increases in sleep efficiency and decreases in wakefulness (p < 0.01), similar to that reported in OSA patients. The REM rebound during recovery, however, could not be attributed to overall REM deprivation since the amount of REM sleep during the OSA phase was not different from the control phase (p = 0.708). This finding suggests that REM rebound during recovery from OSA is not the result of an overall REM sleep deficit per se. Rather, repeated sleep disruption due to the effects of repetitive apneas and hypoxia may lead to an increased REM sleep drive that manifests itself as a REM sleep rebound during recovery sleep after OSA. PMID- 9871948 TI - Cognitive function following acute sleep restriction in children ages 10-14. AB - STUDY OBJECTIVES: Various aspects of human performance were assessed in children after sleep loss. PARTICIPANTS: Sixteen children (7 males, 9 females) between the ages of 10 and 14 years. DESIGN AND INTERVENTIONS: Children were randomly assigned to either a control (CTRL) group, with 11 hours in bed, or an experimental sleep restriction (SR) group, with 5 hours in bed, on a single night in the sleep laboratory. MEASUREMENTS: Both groups were evaluated the following day with a battery of performance and sleepiness measures. Psychomotor and cognitive performance tests were given during four 1-hour testing sessions at 2 hour intervals. RESULTS: A multiple sleep latency test (MSLT) documented shorter latencies for SR children than controls. Significant treatment differences were discovered in three of four variables of verbal creativity, including fluency, flexibility, and average indices. There were also group differences found on the Wisconsin Card Sorting Test (WCST), which may be indicative of difficulty learning new abstract concepts. Measures of rote performance and less-complex cognitive functions, including measures of memory and learning and figural creativity, did not show differences between groups, perhaps because motivation could overcome sleepiness-related impairment for these tasks. CONCLUSIONS: Higher cognitive functions in children, such as verbal creativity and abstract thinking, are impaired after a single night of restricted sleep, even when routine performance is relatively maintained. PMID- 9871949 TI - Adolescent sleep patterns, circadian timing, and sleepiness at a transition to early school days. AB - STUDY OBJECTIVES: This study examined effects on adolescent sleep patterns, sleepiness, and circadian phase of a school transition requiring an earlier start. DESIGN AND SETTING: Adolescents were evaluated in 9th and 10th grades; school start time in 9th grade was 0825 and in 10th grade was 0720. Assessments at each point included 2 weeks of actigraphy and sleep diaries at home, followed by a 22-hour laboratory evaluation, including evening saliva samples every 30 minutes in dim light for determination of dim-light salivary melatonin onset phase (DLSMO), overnight sleep monitoring, and multiple sleep latency test (MSLT). PARTICIPANTS: Twenty-five females and 15 males, ages 14 to 16.2 were enrolled; 32 completed the study in 9th grade and 26 completed in 10th grade. INTERVENTIONS: Participants kept their own schedules, except that laboratory nights were scheduled based upon school-night sleep patterns. MEASUREMENTS AND RESULTS: According to actigraphy, students woke earlier on school days in 10th than in 9th grade, but they did not go to sleep earlier and they slept less. DLSMO phase was later in 10th grade (mean = 2102) than 9th grade (mean = 2024). Sleep latency on MSLT overall was shorter in 10th (mean = 8.5 minutes) than in 9th (mean = 11.4 minutes), particularly on the first test of the morning at 0830 (5.1 vs 10.9 minutes). Two REM episodes on MSLT occurred in 16% of participants in 10th grade; one REM episode occurred in 48%. When those with REM sleep on one or both morning MSLTs (n = 11) were compared to those without morning REM, significant differences included shorter sleep latency on the first test, less slow wave sleep the night before, and later DLSMO phase in those who had morning REM. CONCLUSIONS: Early start time was associated with significant sleep deprivation and daytime sleepiness. The occurrence of REM sleep on MSLT indicates that clinicians should exercise caution in interpreting MSLT REM sleep in adolescents evaluated on their "usual" schedules. Psychosocial influences and changes in bioregulatory systems controlling sleep may limit teenagers' capacities to make adequate adjustments to an early school schedule. PMID- 9871950 TI - Analysis of clustered data in receiver operating characteristic studies. AB - Clustered data is not simply correlated data, but has its own unique aspects. In this paper, various methods for correlated receiver operating characteristic (ROC) curve data that have been extended specifically to clustered data are reviewed. For those methods that have not yet been extended, suggestions for their application to clustered ROC studies are provided. Various methods with respect to their ability to meet either of two objectives of the analysis of clustered ROC data are compared to consider a variety of ROC indices and their accessibility to researchers. The available statistical methods for clustered data vary in the range of indices that can be considered and in their accessibility to researchers. Parametric models permit all indices to be considered but, owing to computational complexity, are the least accessible of available methods. Nonparametric methods are much more accessible, but only permit estimation and inference about ROC curve area. The jackknife method is the most accessible and permits any index to be considered. Future development of methods for clustered ROC studies should consider the continuation ratio model, which will permit the application of widely available software for the analysis of mixed generalized linear models. Another area of development should be in the adoption of bootstrapping methods to clustered ROC data. PMID- 9871951 TI - Correcting for verification bias in studies of a diagnostic test's accuracy. AB - Standard methods for assessing the accuracy of diagnostic tests require determination of true disease status for each study patient. In practice, some study patients might not have verified disease status. If the decision to verify a patient is influenced by the test result, analysis using only verified cases might lead to biased results, commonly known as verification bias. This paper reviews recent developments in bias-correction methods for studies on the accuracy of diagnostic tests. PMID- 9871952 TI - Evaluation of diagnostic tests without gold standards. AB - This paper reviews statistical methods developed to estimate the sensitivity and specificity of screening or diagnostic tests when the fallible tests are not evaluated against a gold standard. It gives a brief summary of the earlier historical developments and focuses on the more recent methods. It covers Bayesian approaches and longitudinal studies with repeated testing. In particular, it reviews the procedures that do not require the assumption of independence between tests conditional on the true disease status. PMID- 9871953 TI - Sample size calculations in studies of test accuracy. AB - Methods for determining sample size for studies of the accuracy of diagnostic tests are reviewed. Several accuracy indices are considered, including sensitivity and specificity, the full and partial area under the receiver operating characteristic curve, the sensitivity at a fixed false positive rate, and the likelihood ratio. Sample size formulae are presented for studies evaluating a single test and studies comparing the accuracy of tests. Four real examples illustrate the concepts involved in sample size determination. Lastly, various study design issues are discussed, such as sampling methods, choices in format for the test results, and the issue of replicated readings. PMID- 9871955 TI - Prolonged nasal shedding and viraemia of cytopathogenic bovine virus diarrhoea virus in experimental late-onset mucosal disease. AB - A calf persistently infected with bovine virus diarrhoea virus (BVDV) was super infected with a heterologous BVDV strain, C874, which contained non cytopathogenic and cytopathogenic viruses. High titres of cytopathogenic BVDV were recovered in the three to four weeks after the challenge. Thereafter low titres of cytopathogenic virus were recovered repeatedly from the blood and the nose, with the titres in nasal secretions increasing in the four weeks before the onset of clinical signs. Neutralising antibodies against the challenge cytopathic virus (C874cp) were first detected 21 days after the super-infection, but these antibodies failed to neutralise the persisting non-cytopathogenic and cytopathogenic viruses isolated from the animal during the course of the infection. Serum collected from 105 days after the super-infection neutralised the cytopathogenic viruses isolated on day 105 and postmortem. These data indicate that unaltered wild-type C874cp was not directly responsible for the late-onset mucosal disease. PMID- 9871954 TI - Assessment of the injuries sustained by pregnant sows housed in groups using electronic feeders. AB - The incidence of injuries to pregnant sows kept in dynamic groups in a commercial herd was recorded over a period of 18 months. The sows were maintained in a solid floored building with a straw-bedded lying area allowing 1.7 m2 per sow and fed by means of electronic feeders. Injuries on 12 possible areas of the body were assessed every week, and graded on the scale of 0 to 3 (maximum body score 36). The lesion scores sustained by the sows were very low, with a maximum of 19 and a median of 4. Lesions of the greatest severity constituted only 0.16 per cent of all the recorded injuries. The injuries were not distributed uniformly over the body, most being on the shoulders, flanks and hindquarters, with very few on the head or in the vulval region. The total injury score was related to parity, with sows in parities one, two and three sustaining more injuries than sows in later parities, and it was also related negatively to bodyweight. More agonistic interactions were observed between sows in the lying area and in the vicinity of the feeders than in the dunging area. PMID- 9871956 TI - Caesarean section in a pygmy hippopotamus (Choeropsis liberiensis) and the management of the wound. AB - A laparohysterotomy was performed on a 17-year-old pygmy hippopotamus (Choeropsis liberiensis) to remove a dead over-sized fetus. The operation was successful and the animal recovered, but the surgical wound broke down and healed slowly by second intention. PMID- 9871957 TI - Bottlenosed dolphins (Tursiops truncatus) as a possible cause of acute traumatic injuries in porpoises (Phocoena phocoena). PMID- 9871958 TI - Fertility of repeat breeder cows in subsequent lactations. PMID- 9871959 TI - Salter-Harris type III fracture of the proximal femoral physis in a puppy. PMID- 9871960 TI - Treatment of swine dysentery--problems of antibiotic resistance and concurrent salmonellosis. PMID- 9871961 TI - Evidence-based medicine. PMID- 9871962 TI - Control of slugs and snails. PMID- 9871963 TI - [Development of preventive iodine administration in Austria]. PMID- 9871964 TI - [Risks and minimizing risks in forensic leave of absence--the forensic psychiatric viewpoint]. PMID- 9871965 TI - [New aspects in treatment of systemic mycoses]. AB - The incidence of systemic fungal infection has been increasing during the last two decades. Candida and Aspergillus spp. are the classical opportunistic pathogens. Rare fungi, such as Mucor, Rhizopus, Fusarium, Trichosporon, Paecilomyces, Alternaria, Cladosporium and Pseudoallescheria, are emerging as cause of systemic fungal infection in the immunocompromised host. For more than 40 years Amphotericin B has been the gold standard of antifungal treatment because of its broad spectrum comprising yeasts, dimorphic fungi and moulds. Its nephrotoxicity has led to the development of lipid-associated preparations of amphotericin B: liposomal amphotericin B, amphotericin B colloidal dispersion and amphotericin B lipid complex. These preparations are less nephrotoxic, but higher doses than those of conventional amphotericin B are needed to achieve the same effect. The triazole fluconazole is the treatment of choice in infections caused by Candida albicans. New antifungal compounds are voriconazole and the candins, the pradimicin/benanomycin family, nikkomycin Z and a liposomal preparation of nystatin. PMID- 9871966 TI - [Preventive iodine administration and iodine excretion in the Vienna area and in the forest quarter]. AB - In order to reduce the still substantial iodine deficiency in the Austrian population, compulsory iodisation of salt was increased in 1990 from 10 mg potassium iodide/kg salt to 20 mg potassium iodide/kg. In this investigation we evaluated the adequacy of iodine supply in Vienna and the Waldviertel, a rural region northwest of Vienna. Daily iodine excretion (which reflects daily iodine intake) was investigated in 92 persons from the Waldviertel (all without thyroid gland pathology) and 110 persons from Vienna (54 with unremarkable thyroid glands, 56 with endemic goiter). Daily iodine excretion was higher in persons from the Waldviertel (161 +/- 90.7 micrograms/24 h, p < 0.05) than in those from Vienna (with healthy thyroid glands 126.4 +/- 42.9 micrograms/24 h and with goiter 117.2 +/- 60.5 micrograms/24 h, resp.). In both populations iodine supply as defined by the WHO (excretion of > 150 ug iodine/day) was inadequate. The recommended level was not achieved in 50% of the persons from the Waldviertel region and in 75% of persons from Vienna (healthy thyroid glands 76%, goiter: 75%). Iodine deficiency (intake < 100 micrograms/24 h) was present in 42% persons from the Waldviertel and in 31% and 24% persons, respectively, from Vienna with unremarkable thyroid glands and goiter. We conclude that although the iodine content of salt was increased, an iodine deficiency was present in a considerable portion of the population of an industrialized country (approximately 40% of persons from a rural region and 30% from the city of Vienna). PMID- 9871967 TI - [Value of drug screening in granting leave of absence to prisoners]. AB - Permitting prisoners to leave the prison for a certain time is one basic element of rehabilitation. As a rule, the inmate is not allowed to leave the prison before a psychologist or psychiatrist is consulted. A considerable number of criminals have committed crimes under the influence of addictive drugs. Many of them continue to indulge in drug abuse even in prison. Therefore, a special programme was introduced in the prison of Graz-Karlau in Austria. In the course of this programme, inmates who are about to leave prison are made to undergo a urinarlysis with immunological methods. In 1997, positive results were registered in 95 of 678 tested prisoners. Thus, this method is an effective means of determining whether prisoners may be granted a temporary release. Persons with a positive urine test may be refused permission. At the same time, every inmate with a positive test may avail himself of an appropriate treatment programme. PMID- 9871968 TI - [Stress fractures of the lower leg and foot]. AB - Stress fractures are caused by overstrain. Depending on their etiology, stress fractures may be categorized as insufficiency or fatigue fractures. A fatigue fracture occurs when repeated axial or rotational force is applied to the bone. In contrast, an insufficiency fracture is observed in bones with altered biomechanical characteristics. In a mixed patient population drawn from the Military Hospital Graz and the Unfallkrankenhaus (Traumatological Hospital) Graz, we were able to corroborate these etiologies. At the former institution, all 21 recruits (mean age, 20.5 years) had fractures of the fatigue type. In the population at the Traumatological Hospital--16 patients with a higher average age of 43.5 years--insufficiency fractures predominated. Diagnosis and differential diagnosis proved difficult in both groups, since plain radiography does not demonstrate the fracture for 2-3 weeks after the occurrence. All stress fractures were treated conservatively. Approximately two thirds of all stress fractures occur in the lower extremities. The main age group at risk is unknown. A sudden change in the activity or training regimen or a change in biomechanics can cause a stress fracture. The absence of individual risk factors makes it difficult to prevent stress fractures. PMID- 9871970 TI - [Arteriovenous fistula of the lung: a rare cause of erythrocytosis]. AB - Various chronic pulmonary diseases can cause hypoxia mediated erythrocytosis. We report on a 46 year old male patient presenting with erythrocytosis, in whom a pulmonary arteriovenous fistula on the basis of a vascular malformation was identified as a rare cause of hypoxic erythrocytosis. Thus, congenital pulmonary vascular malformations can become clinically manifest in advanced age. PMID- 9871969 TI - [Interscalene plexus block for mobilizing chronic shoulder stiffness]. AB - INTRODUCTION: After mobilisation of a frozen shoulder, patients should work with a physiotherapist one to two times a day. In the remaining time patients must exercise on their own: otherwise, the shoulder is likely to become less mobile. Exercise is only possible in the absence of pain. Therefore, adequate analgesic therapy for four to five days after the first mobilisation is of great importance. We used interscalene blockade of the brachial plexus to achieve mobilisation in patients suffering from a painful frozen shoulder. We investigated the efficacy of this technique by registering the use of additional analgesics and documenting the patient's assessment. The observation time was 18 months. METHODS: Thirty-seven patients were treated with a plexus block using a catheter inserted at the affected side. Catheter location was verified with electric nerve stimulation. In all patients a bolus of 20 ml bupivacaine 0.375% was given for initial mobilisation, followed by 10 ml bupivacaine 0.25% every 6 hours. RESULTS: In 34 of 37 (92%) patients, the shoulder could be mobilised without general anaesthesia. Thirty-three of 37 (89%) patients did not need any additional analgesics during the following mobilizations. The mean duration of catheter treatment was 4 days; no infections occurred. The following side effects were observed at the first mobilisation: Horner's syndrome in 20 patients (54%), paresis of the recurrent nerve in 9 patients (24%) and paresis of the phrenic nerve in 6 patients (16%). During subsequent treatment, Horner's syndrome was observed in 15 patients (41%) and a paresis of the recurrent nerve in 8 patients (22%). In an interview after the end of the treatment period all patients stated that they would use this method again if necessary. CONCLUSIONS: With the use of this regional anaesthesia technique it was possible to provide comfortable pain therapy during the mobilisation period. In order to achieve sufficient analgesia with only a mild motor blockade of the motoneurons, the exact location of the catheter tip is important. The latter can be verified by electrical stimulation. PMID- 9871971 TI - Rapid enzyme immunoassay for measurement of bovine progesterone. AB - Reproductive management is a primary financial concern of the dairy industry with missed estrus detection one of the major causes of lost income. A rapid enzyme immunoassay (EIA) was developed for on-line measurement of progesterone in bovine milk with a biosensor for detection of estrus. The EIA was developed using covalent binding microtiter wells, monoclonal antibody, horseradish peroxidase, and 3,3',5,5'-tetramethylbenzidine (TMB). The EIA took 8 min and had a dynamic response for progesterone in buffer and milk between 0.2 and 20 ng/ml. PMID- 9871973 TI - Novel integrated optical sensor based on a grating coupler triplet. AB - A novel approach for accomplishing robust integrated optical biosensors is presented. The principle is based on a symmetric grating coupler structure with the inherent feature of compensating disturbances originating from different kinds of mechanical instabilities. The complete sensor system has no moving parts and provides the high sensitivities typical for integrated optical sensors based on grating couplers. The feasibility of this approach is demonstrated by determining the refractive index of liquids as well as by measuring the specific binding of biomolecules (anti-chicken IgG) to immobilized chicken IgG on the sensor chip surface. PMID- 9871972 TI - Biosensor for on-line measurement of bovine progesterone during milking. AB - Reproductive management is a major financial concern of the dairy industry, with missed estrus detection a main cause of lost income. A biosensor was developed for on-line measurement of progesterone in bovine milk and detection of estrus. The biosensor used an enzyme immunoassay format for molecular recognition, which was developed to run in approximately eight minutes. The sensor was designed to operate on-line in a dairy parlor using microinjection pumps and valves for fluid transport, fiber optics and photodiodes for light measurement, and a control computer for sequencing. Calibration showed a dynamic response between 0.1 and 5 ng/ml progesterone in milk. The reusability of the test well was evaluated. Thiocyanate (0.5 M, pH 5.1) quickly regenerated the antibody surface while maintaining antibody activity for 15-20 cycles, but noise from the residual enzyme limited reusability. PMID- 9871974 TI - In vivo nitric oxide sensor using non-conducting polymer-modified carbon fiber. AB - Nitric oxide (NO) is emerging as a very important and ubiquitous gaseous messenger in the body. The response characteristics of NO sensors made of non conducting polymer modified carbon fiber electrodes are investigated to determine their selectivity, sensitivity, and stability for in vivo use. A composite polymer, comprising Nafion, m-phenylenediamine, and resorcinol, showed the best selectivity and stability to amperometric NO detection. The non-conducting, self limiting polymer film protects the electrode from interference and fouling by other biochemicals. Although the relative sensitivity to NO of the modified sensor is lower than that of the unmodified carbon fiber electrodes (less than 6%), the composite polymer electrode showed high selectivity against ascorbic acid (> 2000:1), nitrite (> 600:1), and dopamine (> 200:1). The stability of the NO sensor was maintained for at least 1 week. The NO sensitivity after in vivo experiments (n = 8) is 88.1 +/- 5.6% of initial sensitivity data obtained before in vivo experiments. Preliminary in vivo experiments done with this electrode are shown to capture elevated NO levels in brain following an ischemic injury. PMID- 9871975 TI - In situ mapping of community-level cellular response with catalytic microbiosensors. AB - Chemotaxis, the migration of cells in the direction of a spatial chemical gradient, is important in disease progression, microbial ecology, and bioremediation. The ability to map chemoattractant gradients and the corresponding cellular growth and motility patterns is essential to the study of chemotaxis. Microelectrodes and microbiosensors have the potential to measure chemoattractant gradients with high spatial resolution. In this study, Clark-type amperometric microelectrodes and microbiosensors were used to measure solute concentrations gradients generated by a chemotactic band of Escherichia coli in a semi-solid gel. A computerized image analysis system was used to simultaneously measure the cellular concentration profile across the chemotactic band. The experimental results compared favorably with a mathematical model of solute and cell transport in the gel. Scanning electron micrographs (SEM) of micro(bio)sensor tips taken after 6 months of use showed evidence of degradation, including adhesion of foreign particles to the glass body, the adhesion of a small gel capsule to the sensor tip, and separation of the bio-interface from the tip. A needle-type microbiosensor was constructed to better protect the tip and hence increase the ruggedness of the microbiosensors. PMID- 9871976 TI - Improved operational stability of biosensors based on enzyme-polyelectrolyte complex adsorbed into a porous carbon electrode. AB - A novel porous active carbon is utilized in order to adsorb the diethylaminoethyl dextran (DEAE-dextran)-enzyme stabilized complexes, for the construction of highly stable biosensors. The interaction of DEAE-dextran with the examined enzymes increases dramatically the operational stabilization of the sensors, without adverse effects on the enzyme activity. At the same time, the porous active carbon allows for high enzyme loading, good electrical contact and low resistance throughout the sensing element. Glucose oxidase and horseradish peroxidase are used as model enzymes in this study to construct biosensors, with very good reproducibility (less than 5% RSD). As a result, the glucose sensor exhibits very long operational stability (over a period of 5 months), while the hydrogen peroxide sensor retains its initial activity after several weeks. PMID- 9871977 TI - Covalent coupling of immunoglobulin G to self-assembled monolayers as a method for immobilizing the interfacial-recognition layer of a surface plasmon resonance immunosensor. AB - Protocols have been developed for the random and site-directed covalent coupling of immunoglobulin G [anti-hIgG] [IgG] to silver surfaces modified with a self assembled monolayer [SAM] of thioctic acid, mercaptopropionic acid [MPA], L cysteine or 4-aminothiophenol [PATP]. A surface plasmon resonance [SPR] immunosensor fabricated with a more ordered and hydrophilic IgG-SAM-silver interfacial layer, demonstrates an increased ability for performing sensitive and selective assay of human immunoglobulin G [hIgG] compared with a device fabricated with a physically-adsorbed IgG-silver interfacial-layer due to reduced levels of non-specific binding. Detection limits [DL] for hIgG from serum down to 6 micrograms/ml (40 nM) and assay sensitivities up to 0.24 ng hIgG/mm2/nM are reported. PMID- 9871978 TI - Application of Teflon-AF thin films for bio-patterning of neural cell adhesion. AB - Teflon AF, an amorphous copolymer of polytetrafluoroethylene (PTFE) with 2,2 bis(triflouromethyl)-4,5-difluoro-1, 3-dioxole, has been receiving widespread attention in the opto-electronics industries and elsewhere for its superior optical and dielectric properties. The objective of the present study was to investigate surface parameters that may be required for the application of thin films of Teflon AF in the fields of biomaterials and bioelectronics. Using standard microelectronics procedures, micro-patterned thin films of Teflon-AF were fabricated, and their surface properties monitored and optimized with the aid of highly surface sensitive spectrometric techniques. Finally, their capability to inhibit or bio-pattern cell adhesion was tested with various neural cell lines. PMID- 9871979 TI - Electron transfer kinetics at a biotin/avidin patterned glassy carbon electrode. AB - Photolithographic techniques using a laser interference pattern were used to attach photobiotin to micron-sized stripes on the surface of a carbon electrode. Fluorophore-tagged avidin was attached to this spatially-patterned biotin with essentially no loss in spatial resolution. The kinetics of the glassy carbon surface were examined to see if electron transfer sites could indeed be segregated from the attachment sites of photobiotin-immobilized avidin. The ECL of luminol and SECM were used to verify the segregation between underivatized sites (which exhibit normal electron transfer kinetics) and extensively derivatized biotin/avidin surfaces (which presumably exhibit slow electron transfer kinetics). Both techniques were found to be capable of differentiating the protein-covered surface from bare carbon with sufficient resolution to tell whether a significant portion of the carbon surface is still active and available to detect the product of an enzyme generated analyte. These results indicate that extensive biotin/avidin derivatization of the surface does decrease the electron transfer rate of a carbon electrode, and that the photolithographic approach was able to modify specific sections of the electrode surface, while leaving other regions untouched and available for facile electron transfer. This leads to a more general protocol for the construction of enzyme-based biosensors which utilize diffusable mediators. PMID- 9871980 TI - Infrared spectroscopy of glycolipids. AB - The application of infrared spectroscopy to the physicochemical characterization of glycolipids as biologically important membrane constituents is described. In this contribution, the analysis of diacyl- and sphingosine-based (cerebroside, ganglioside) oligosaccharides and the class of lipid A-anchored lipooligo- and polysaccharides is reviewed. Furthermore, interaction of glycolipids with various agents as well as of sugars with membranes are discussed. The reviewed data prove the capacity of FTIR spectroscopy to monitor intra- and intermolecular interactions under physiologically relevant conditions refering to pH, temperature, and ion concentrations. This includes the characterization of acyl chain order and the beta<-->alpha chain melting transition, cation and drug binding to charged groups, and supramolecular organization of the glycolipid aggregates. In various examples, the biological relevance of IR data analysis are presented. PMID- 9871981 TI - The use of FT-IR for quantitative studies of the apparent pKa of lipid carboxyl groups and the dehydration degree of the phosphate group of phospholipids. AB - Fourier-transform infrared spectroscopy (FT-IR) has been applied to the quantitative study of the dehydration of the phosphatidylserine phosphate group in the presence of Ca2+ exerted by different molecules, such as diacylglycerol, sphingosine and stearylarnine, by using a partial least-squares statistical procedure. By using this method it was observed that diacylglycerol enhanced the dehydration of this PO2- group produced by Ca2+ whereas the amino-bases sphingosine and stearylamine protected the phosphate group from the dehydration produced by Ca2+ due to the very strong electrostatic interaction established. The apparent pKa of lipid carboxyl groups can also be estimated by using FTIR. The method consisted in quantifying the absorbance intensities due to the protonated and the unprotonated forms of the specific group being studied. The pKa of the carboxyl group of [1-13C]-palmitic acid included in dipalmitoylphosphatidylcholine membranes was found to be 8.7, a value much higher than that estimated from a molecular solution of the fatty acid. It was observed using the same method that the pKa of free fatty acids in model stratum corneum lipid mixtures was in the range 6.2-7.3 increasing with the preponderance of oleic acid over palmitic acid. Finally the pKa of the carboxyl group of phosphatidylserine was shifted from 4.6 in the pure phospholipid to 2.1 and 2.2 in the presence of equimolar sphingosine and stearylamine, respectively, as a consequence of electrostatic interactions. PMID- 9871982 TI - Infrared studies of protein-induced perturbation of lipids in lipoproteins and membranes. AB - The paper reviews the main recent publications concerning infrared (IR) spectroscopy as applied to the study of lipid-protein interactions in model and cell membranes, lipoproteins, and related systems (e.g. lung surfactant). The review focuses mainly on transmission IR. Based on the available data, a number of general conclusions are presented on the perturbations caused by proteins on either the hydrocarbon chains, the polar headgroups or the interface region. Lipid-protein interactions in native cell membranes do not reveal significant differences from what is observed in semisynthetic model systems. PMID- 9871983 TI - Infrared spectroscopy of supported lipid monolayer, bilayer, and multibilayer membranes. AB - Cecropin A was examined in supported monolayer, bilayer, and multibilayer lipid membranes using attenuated total internal reflection Fourier-transform infrared spectroscopy. The spectral features provide an abundance of information about the conformation and orientation of the peptide, as well as about the effects of the peptide on lipid order. In this case, they serve to contrast results from the three preparations. The results of monolayer and bilayer studies are generally similar, although differences in the nature of the membranes appear to cause minor changes in the conformation and orientation of the peptide. The results of the multibilayer studies are different in many respects from those of the monolayer and bilayer studies, suggesting that fundamentally different peptide lipid interactions occur in multibilayers. PMID- 9871984 TI - On the use of deuterated phospholipids for infrared spectroscopic studies of monomolecular films: a thermodynamic analysis of single and binary component phospholipid monolayers. AB - A thermodynamic study of monolayer mixing behavior was performed using binary lipid mixtures of DPPC + DOPG (7:1 mol:mol) as well as a mixture containing the phosphocholine acyl chain perdeuterated analog of DPPC, DPPC-d62 (i.e. 7:1 mol:mol DPPC-d62 + DOPG). An analysis was performed on these isotherms that calculated the Helmholtz excess free energy of mixing to determine potential thermodynamic differences in the fully protiated DPPC:DOPG monolayer versus the identical monolayer substituted with acyl chain perdeuterated DPPC (i.e. DPPC d62). The main conclusions of this study are that: (1) the temperature-dependent thermodynamic phase behavior of DPPC-d62 differs substantially from that of DPPC, and these differences appear magnified when these molecules are studied as monomolecular films at the A/W interface as opposed to bulk phase systems; and (2) the calculated excess free energy of mixing values, deltaFxs(A), for the deuterium-containing 7:1 DPPC-d62:DOPG binary monolayer film show a type of near ideal mixing behavior above 20 degrees C that is consistent with the 'squeezing out' of the DOPG component in the binary mixture containing the DPPC-d62 component. This type of behavior does not occur in the graph of the excess free energy of mixing for the fully protiated 7:1 DPPC:DOPG binary monolayer film. The use of acyl chain perdeuterated DPPC (i.e. DPPC-d62) in binary monomolecular mixtures with other phospholipids produces a real and measurable difference on the thermodynamic properties of the monolayer when compared to the case of the fully protiated monomolecular film. In particular, the presence of DPPC-d62 in a 7:1 mol:mol DPPC-d62:DOPG binary monomolecular film may over-state the fluid nature of the monolayer at any given temperature, thereby leading to an over estimate of the amount of material potentially 'squeezed out' of the monomolecular film. These results have implications for a commonly used IR spectroscopic method that relies on the incorporation of deuterium-labeled phospholipids into monolayers for vibrational spectroscopic analysis of individual components in a multicomponent monomolecular film. PMID- 9871986 TI - The effect of magnesium ions during beer fermentation. AB - When cells of Saccharomyces cerevisiae ale strain C1028 were grown aerobically, ethanol production displayed a hyperbolic increase over a limited range of magnesium concentrations up to approximately 0.7 mM. Entry of cells into the stationary growth phase and the time of maximum ethanol and minimum sugar concentrations correlated with a period of maximum Mg2+ concentration in the growing media. It is suggested that magnesium accumulation by yeast cells may be usefully exploited in biotechnology concerned with the production of beer. PMID- 9871985 TI - Vibrational spectroscopic studies of lipid domains in biomembranes and model systems. PMID- 9871988 TI - Estimation of stature from somatometry of skull. AB - In order to investigate the possibility of estimating stature from somatometry of the skull, we carried out a study on 124 Japanese cadavers (comprising 77 males and 47 females) that had been autopsied at our laboratory between July 1986 and June 1991. Somatometry of the skull was performed on diameter (distance between glabella and external protuberance) and circumference (length around the skull through two points: the glabella and the external protuberance). The regression equations calculated were as follows: stature in males = (diameter + circumference) x 1.35 + 70.6 (standard error of estimate (S.E.) = 6.96 cm); stature in females = circumference x 1.28 + 87.8 (S.E. = 6.59 cm); stature in both sexes = (diameter + circumference) x 1.95 + 25.2 (S.E. = 7.95 cm). These S.E.s appear to be larger than those obtained for other parts of the body. However, in cases where identification is required by means of only the skull, this method could prove useful. PMID- 9871987 TI - A blue encrustation found on skeletal remains of Americans missing in action in Vietnam. AB - A blue encrustation was found on the repatriated remains of three U.S. Servicemen listed as missing in action (MIA) from Vietnam after 28 years. The identification and origin of the blue material was determined. Scanning electron microscopy with energy dispersive analysis and powder X-ray diffraction identified the material as the mineral vivianite, Fe3 (PO4)2.8H2O. Vivianite has been found often associated with fossilized bone and teeth, but this example is unusual in that it is only the second published forensic example of vivianite growing from human bone after such a short period of time. The presence of vivianite provides information leading to a more complete and accurate understanding of the taphonomic process associated with American MIA remains. PMID- 9871989 TI - Solid-phase microextraction (SPME) of drugs and poisons from biological samples. AB - Solid-phase microextraction (SPME), a new solvent-free sample preparation technique, was invented by C. Arthur and J. Pawliszyn in 1990. This method mainly was applied for the extraction of volatile and semi-volatile organic pollutants in water samples. However, since 1995, SPME has been developed to various biological samples, such as whole blood, plasma, urine, hair and breath, in order to extract drugs and poisons in forensic field. The main advantages of SPME are: high sensitivity, solventless, small sample volume, simplicity and rapidity. We have reviewed the papers published in recent years about SPME in biological samples, and sorted out main experimental conditions, such as fibers, matrixes, the extraction approaches and time, as well as the acceleration method. We would expect SPME technique to have a promising future for toxicological analysis in forensic practice. PMID- 9871990 TI - Immunohistochemical detection of fibronectin in postmortem incised wounds of porcine skin. AB - Fibronectin plays an important role in tissue repair and wound healing. Previous literature reports indicated that fibronectin could be a marker of vitality for wounds with a survival time of more than a few minutes. In order to verify these findings were performed immunohistochemical investigations on the expression of fibronectin in incised wounds of porcine skin inflicted into various anatomical regions in the early postmortem interval (0-5 min after circulatory arrest). Tissue specimens were collected 12-14 h postmortem and investigated for fibronectin on paraffin sections (alkaline phosphatase and monoclonal anti alkaline phosphatase procedure). A markedly positive expression of fibronectin (not only next to the wound margin and clearly stronger than in normal undamaged skin) could be demonstrated in eleven out of 36 samples. A moderately positive fibronectin reaction was detectable in seven further specimens. Muscular contraction bands were observed in nine out of 36 specimens. The various reaction patterns of fibronectin cannot be finally explained, but are probably due to a passive transudation of skin tissue by blood components from injured vessels. As a result of these investigations the specificity and validity of fibronectin as a parameter of vitality has to be questioned. Fibronectin immunohistochemistry again reveals that the validity of parameters as vital criteria decreases when their manifestation time is short; postmortem/supravital induction seems to be possible resembling vital reactions in these cases. PMID- 9871991 TI - Analytical toxicology of fluorinated inhalation anaesthetics. AB - The chemical and pharmacological properties of the current fluorinated inhalation anaesthetics, halothane, enflurane, isoflurane, sevoflurane and desflurane, are surveyed with implications to toxicity. Analytical methods, especially gas chromatography with head space, purge and trap, or pulse heating extraction, are reviewed in forensic toxicological and occupational/therapeutic monitoring contexts. PMID- 9871992 TI - Mannan binding lectin and the sudden infant death syndrome. AB - Mannan binding lectin (MBL) may be important for innate immunity and some cases of sudden infant death syndrome (SIDS) may be preceded by bacterial infection. Therefore, relative MBL deficiency might be associated with susceptibility to SIDS. We measured MBL concentrations in 46 SIDS infants and 26 controls. The proportion of subjects with low MBL values was similar in the two groups. However, the mean for the SIDS group (3 micrograms/ml) was higher than that of the controls (2.2 micrograms/ml; P < 0.05). We interpret this difference as due to acute phase responses and suggest these findings are consistent with the view that some cot deaths are preceded by bacterial infections. PMID- 9871993 TI - Identification of Cannabis pollens using an allergic patient's immunoglobulin E and purification and characterization of allergens in Cannabis pollens. AB - Cannabis pollen allergens were detected using the serum of an allergic patient. The allergens were then purified by sequential column chromatography (including DE52 cellulose and phenyl-Sepharose CL-4B) and preparative HPLC. The molecular weight of the allergens were determined as 10,050 and 13,706 by matrix-assisted laser desorption/ionization time of flight mass spectrometry. We utilised Western blotting and development of an enzyme-linked immunosorbent assay for the detection of Cannabis pollen allergens. PMID- 9871994 TI - Sequence polymorphism of mitochondrial DNA control region in Japanese. AB - Sequence polymorphisms of the mitochondrial DNA (mtDNA) control region, hypervariable regions I and II, from 100 unrelated Japanese were determined by PCR amplification and direct sequencing. Sequences of 404 nucleotides for hypervariable region I and 379 nucleotides for region II were obtained. Variable sites (85 and 45) were revealed in region I and region II, respectively, as compared to the reference sequence, and a total of 96 different genetic patterns from both regions I and II were determined. A point mutation heteroplasmy was observed at the ratio of approximately 50:50 from one individual at the sequence position 151 showing a nucleotide transition from C to T. The probability of identity was estimated as 2.3% for region I, 3.9% for region II, and 1.1% combined for both regions. These results suggest that sequence polymorphism of mtDNA control region would be very useful in forensic practice as a marker for individual identification. PMID- 9871995 TI - Reproducibility of mtDNA analysis between laboratories: a report of the European DNA Profiling Group (EDNAP). AB - The aim of this collaborative exercise was to determine whether uniformity of mtDNA sequencing results could be achieved among different EDNAP laboratories. Laboratories were asked to sequence mtDNAHV1 region (16024-16365) from three bloodstains, proceeding in accordance with the protocol and strategies currently used in each individual laboratory. Cycle sequencing was used by 11 laboratories and solid phase single stranded sequencing was used by one laboratory. Different PCR strategies and PCR conditions were used by the different laboratories. Three laboratories used semi-nested PCR, two nested PCR, three direct amplification of HV1 and four amplification of overlapping fragments covering the HV1 region. Despite the diversity of methodologies used, all the laboratories reported the same results. The successful result of this exercise shows that PCR based mtDNA typing by automated sequencing is a valid, robust and reliable means of forensic identification despite the different strategies and methodologies used by the different laboratories. PMID- 9871996 TI - Drug abuse of Finnish male prisoners in 1995. AB - The purpose of the present research was to estimate the extent and variety of abuse of illegal drugs, use and misuse of hypnotics and sedatives and anabolic steroids in the Finnish prison population. The study was undertaken during October-November 1995 at four prisons, three of which were closed institutions and one an open prison; one of the three closed institutions was a juvenile prison. There was a total of 707 inmates in the prisons selected for the study. Questionnaires were given personally to all prisoners in the open prison and in the young prisoners' division in the juvenile prison, but in two large central prisons only some divisions were selected for the study. The questionnaires were completed by 354 prisoners; 75 prisoners refused to respond. A total of 27.7% of subjects reported taking illegal drugs while in their current prison and 70.1% had sometimes used them. Of those who were drug-free before their first imprisonment, 21.7% began using drugs in prison. At present hypnotics and sedatives were reported as in use by 41.8% of subjects, one-third as prescribed drugs and about 10% illicitly. A total of 3.7% of subjects reported taking anabolic steroids in the current prison. Cannabis and amphetamine were the most common illegal drugs reported. Intravenous drug use was reported by 19.2% of the respondents at some point in their lives, and 10.7% of prisoners had injected drugs in their current prison. Use of illegal drugs and misuse of drugs were significantly higher among young prisoners (< or = 25 years of age). PMID- 9871997 TI - The use of cryopreserved apical protoplasts from Curvularia lunata for electrotransformation. AB - An electroporation method, utilizing cryopreserved protoplasts, has been developed for the steroid 11-hydroxylating fungus Curvularia lunata strain IM 2901. Protoplasts released from the apical parts of 24- and 48-h-old mycelia were suspended in cryopreservation buffer and stored at -75 degrees C for several weeks. The thawed and freshly prepared (control) protoplasts were electroporated with pAN 7-1 plasmid carrying the Escherichia coli hygromycin B resistance gene (hph) under the control of Aspergillus nidulans sequences. The electroporation efficiency of the control protoplasts with plasmid pAN 7-1 was 7.5 and 12.0 transformants per microgram DNA (protoplasts liberated from 24- and 48-h-old mycelia, respectively). Protoplasts released from the younger mycelium were more stable according to their reversion ability to mycelial form and transformation efficiency. After 16 weeks of cryopreservation the yield of electroporation was 61.3% of the control value. All isolated electrotransformants proved to be stable for a period of > 4 months even without selective pressure. PMID- 9871998 TI - Comparative study of two outer membrane protein genes from Porphyromonas gingivalis. AB - A periodontal pathogen, Porphyromonas gingivalis possesses either a 53 kD (Ag53) or a 67 kD (Ag67) outer membrane protein (OMP). Almost all sera from patients with periodontal diseases reacted strongly with either Ag53 or Ag67. In previous work the cloning and sequencing of the 53 kD outer membrane protein gene designated pga53 from P. gingivalis FDC381, was reported and the presence of a gene homologous to pga53 in P. gingivalis ATCC 33277 demonstrated. In the present work this pga53-homologous gene from P. gingivalis ATCC 33277 was isolated and characterized. Nucleotide sequence analysis revealed that this gene encoded Ag67, and the gene was designated pga67. The deduced amino acid sequence and composition of pga67 was similar to the amino acid composition and N-terminal partial sequence of Ag67. An open reading frame of pga67 consisted of 1,692 nucleotides encoded as 564 amino acids, including a 49 amino acid signal sequence. The comparative analysis between pga67 and pga53 revealed that (1) the deduced amino acid sequence showed a 30.1% homology; (2) signal sequence and proline-rich regions at the C-terminus were the most conserved regions; (3) considerable differences were found mainly in the middle part of the OMPs; and (4) obvious differences in the two-dimensional models were evoked. These differences between pga67 and pga53 may explain the antigenic diversity between Ag67 and Ag53 OMPs. PMID- 9871999 TI - Klebsiella pneumoniae clinical isolates: susceptibility to some antibiotics and surface hydrophobicity. AB - The susceptibility of Klebsiella pneumoniae strains isolated from the respiratory tract (55), urinary tract (19) and other human body sites (8) to the antibiotics, amoxycillin/clavulanate, cefuroxime, ceftazidime, gentamicin, norfloxacin, colistine, cotrimoxazole and oxolinic acid, as well as their surface hydrophobicity, were studied. The strains expressed a very high sensitivity to the antibiotics (94.7-100%). The surface hydrophobicity of the strains was evaluated by means of three assays and was minimal. The hydrophobicity manifested by adherence to xylene ranged between 0 and 10% for 96.4% of the respiratory strains, and for 100% of the urinary and other sites. Weak binding of Congo red (10-29 micrograms/10(10) cells) showed 96.4% of respiratory isolates as well as 100% of the urinary and other strains. Results of the salt aggregation test showed that 96.4% of respiratory strains and 100% of urinary and other strains aggregated only at 1.5 M, 2 M or higher concentrations. PMID- 9872000 TI - Appearance of fosfomycin resistant Rahnella aquatilis clinically isolated in Japan. AB - Among recent clinical isolates in Japan, strain CU264 was discovered which formed unusual colonies. This strain was identified as Rahnella aquatilis which is usually found in water. The antibiotic susceptibilities against tetracycline, carbenicillin, chloramphenicol, streptomycin, kanamycin, gentamicin, sulphonamide, neomycin, fosfomycin, rifampicin, norfloxacin and nalidixic acid, were investigated. The result demonstrated that the strain was highly resistant to fosfomycin only. It was further shown that this resistance was transmissible with low frequency to Serratia marcescens whereas it was not transmissible to Escherichia coli. PMID- 9872001 TI - GBV-RNA detection by polymerase chain reaction with several primer pairs. AB - The identification of specific genomic sequences of GB viruses (GBV) has made it possible to utilize the polymerase chain reaction (PCR) for evaluation of the viraemia. Several studies have demonstrated the RNA-GBV presence in sera from different patients amplifying several portions of the virus. In this investigation the PCR results when different regions of GBV (NS3, UTR and putative CORE and E1) were amplified in the same sample. In 245 samples studied there were two (0.8%) discordant results and the NS3 primer showed the greatest sensitivity. The lowest percentage of positivity was obtained with CORE-E1 primers. These results could be because the nucleocapside/E1 region was extremely variable in length and sequences, although degenerated primers and probes were used. Discordances were attributable to laboratory errors, variability in the viral genome, the presence of primer inhibitors in samples or a low viral load. PMID- 9872002 TI - A 'sticky' interhemispheric switch in bipolar disorder? AB - Despite years of research into bipolar disorder (manic depression), its underlying pathophysiology remains elusive. It is widely acknowledged that the disorder is strongly heritable, but the genetics are complex with less than full concordance in monozygotic twins and at least four susceptibility loci identified. We propose that bipolar disorder is the result of a genetic propensity for slow interhemispheric switching mechanisms that become 'stuck' in one or the other state. Because slow switches are also 'sticky' when compared with fast switches, the clinical manifestations of bipolar disorder may be explained by hemispheric activation being 'stuck' on the left (mania) or on the right (depression). Support for this 'sticky' interhemispheric switching hypothesis stems from our recent observation that the rate of perceptual alternation in binocular rivalry is slow in euthymic subjects with bipolar disorder (n = 18, median = 0.27 Hz) compared with normal controls (n = 49, median = 0.60 Hz, p < 0.0005). We have presented evidence elsewhere that binocular rivalry is itself an interhemispheric switching phenomenon. The rivalry alternation rate (putative interhemispheric switch rate) is robust in a given individual, with a test-retest correlation of more than 0.8, making it suitable for genetic studies. The interhemispheric switch rate may provide a trait dependent biological marker for bipolar disorder. PMID- 9872004 TI - Contrast discrimination, non-uniform patterns and change blindness. AB - Change blindness--our inability to detect large changes in natural scenes when saccades, blinks and other transients interrupt visual input--seems to contradict psychophysical evidence for our exquisite sensitivity to contrast changes. Can the type of effects described as 'change blindness' be observed with simple, multi-element stimuli, amenable to psychophysical analysis? Such stimuli, composed of five mixed contrast elements, elicited a striking increase in contrast increment thresholds compared to those for an isolated element. Cue presentation prior to the stimulus substantially reduced thresholds, as for change blindness with natural scenes. On one hand, explanations for change blindness based on abstract and sketchy representations in short-term visual memory seem inappropriate for this low-level image property of contrast where there is ample evidence for exquisite performance on memory tasks. On the other hand, the highly increased thresholds for mixed contrast elements, and the decreased thresholds when a cue is present, argue against any simple early attentional or sensory explanation for change blindness. Thus, psychophysical results for very simple patterns cannot straightforwardly predict results even for the slightly more complicated patterns studied here. PMID- 9872003 TI - The effect of different immune responses on the evolution of virulent CXCR4 tropic HIV. AB - We use mathematical models to determine possible mechanisms contributing to the evolution and rise of virulent CXCR4-tropic HIV in vivo. The models predict that the ability of the virus to specialize on a given target cell type depends on the exact fitness landscape of the viral mutants. Because this fitness landscape varies between people, this may explain why the evolution of fully CXCR4-tropic strains only occurs in about 50% of infected patients. Assuming that CXCR4-tropic HIV may evolve, we investigate the effect of different immune responses on the rise of such virulent strains. If we assume that CXCR4-tropic HIV is more cytopathic than CCR5-tropic virus, virulent CXCR4-tropic mutants remain suppressed at low levels both in the absence of an immune response, and in the presence of responses that act on the virus before integration into the host genome. On the other hand, this difference in cytopathogenicity is reduced by the presence of immune responses acting on infected cells, allowing CXCR4-tropic HIV to coexist with the CCR5-tropic virus. These results may help to interpret experimental data and are discussed with reference to the literature. PMID- 9872005 TI - Ventricular fibrillation: one spiral or many? AB - Ventricular fibrillation is the major cause of sudden cardiac death, the leading cause of death in the industrialized world; however, the mechanisms for its onset are not well understood. To further understand the dynamics of fibrillation at and near its onset, we compared spatial and temporal variability of mean interactivation intervals in a stable canine model for ventricular fibrillation. Temporal variability was very small, suggesting that the relevant physiological parameters remained constant during our experiments. Spatial variability was usually significantly larger and appeared incompatible with the dynamics of a single, meandering spiral wave. This confirmed recent results that a single spiral wave cannot generate ventricular fibrillation. Thus the onset of fibrillation is a multistage process, with spiral-wave breakdown providing a crucial step in the quasi-periodic route to fibrillation. PMID- 9872006 TI - Injury-induced expression of endothelial nitric oxide synthase by glial and microglial cells in the leech central nervous system within minutes after injury. AB - It is known that nitric oxide (NO) is produced by injured tissues of the mammalian central nervous system (CNS) within days of injury. The aim of the present experiments was to determine the cellular synthesis of NO in the CNS immediately after injury, using the CNS of the leech which is capable of synapse regeneration, as a step towards understanding the role of NO in nerve repair. We report that within minutes after crushing the nerve cord of the leech, the region of damage stained histochemically for NADPH diaphorase, which is indicative of nitric oxide synthase (NOS) activity, and was immunoreactive for endothelial NOS (eNOS). On immunoblots of leech CNS extract, the same antibody detected a band with a relative molecular mass of 140,000, which is approximately the size of vertebrate eNOS. Cells expressing eNOS immunoreactivity as a result of injury were identified after freezing nerve cords, a procedure that produced less tissue distortion than mechanical crushing. Immunoreactive cells included connective glia and some microglia. Calmodulin was necessary for the eNOS immunoreactivity: it was blocked by calmodulin antagonist W7 (25 microM), but not by similar concentrations of the less potent calmodulin antagonist W12. Thus in the leech CNS, in which axon and synapse regeneration is successful, an increase in NOS activity at lesions appears to be among the earliest responses to injury and may be important for repair of axons. PMID- 9872007 TI - Parasite-induced risk of mortality elevates reproductive effort in male Drosophila. AB - A trade-off between sex and somatic maintenance is fundamental to life-history theory. Tests of this trade-off usually emphasize deleterious consequences of increased reproduction on life span. Here we show the reverse effect, that reductions in the expected life span elevate sexual activity. Experimentally parasitized male Drosophila nigrospiracula lived shorter lives, but before dying, they courted females significantly more than unparasitized controls. This greater courtship resulted in increased mating speed, and potentially greater reproductive success than parasitized males would have achieved otherwise. The results show that an environmental reduction in life span increases reproductive effort, and support the hypothesis of a trade-off between these key life-history traits. PMID- 9872009 TI - [Importance of recombinant proteins in the adjuvant treatment of cancer]. AB - The clinical application of hematopoietic growth factors, which decrease the duration (and severity) of chemotherapy-induced myelosuppression, was made possible by using genetically-modified micro-organisms to produce them (e.g. epoietin). The interest of these products is double: in attenuating the myelosuppression, they decrease the risk of severe infectious complications; the better hematologic tolerance allows to deliver chemotherapy in higher doses that maximize tumoricidal activity. PMID- 9872008 TI - Evolutionary change in a receiver bias: a comparison of female preference functions. AB - Female poeciliid fishes of the sister genera Xiphophorus and Priapella share a preference for males with swords, despite phylogenetic information suggesting that swords arose in Xiphophorus after the divergence of the two genera. This study examines the strength of sword and body-size preferences in a representative of both genera. A comparison of the preference functions reveals that the strength of the preference favouring a sword in P. olmecae is significantly stronger than that in X. helleri. This result demonstrates that the pre-existing bias is not evolutionarily fixed, and that there has been change in the bias favouring the sword, in either the Priapella lineage, or the Xiphophorus lineage, or in both. Although females in both species prefer conspecific males with swords, only X. helleri females also demonstrate a body-size preference. The preference functions for body size and sword length for X. helleri are not significantly different, whereas in P. olmecae the preference function for sword length is significantly stronger than for body size. These combined results indicate that an ancestral bias for body size cannot alone explain the pre existing bias favouring a sword in P. olmecae. PMID- 9872010 TI - [Enzymatic resistance to imipenem in gram-negative bacilli: NMC-A, an original carbapenemase]. AB - Carbapenems such as imipenem are beta-lactam antibiotics usually stable to the hydrolytic activity of beta-lactamases. Nevertheless, resistance to imipenem by production of carbapenemases has been described in few species of Gram negative bacilli. This mechanism of resistance remains exceptional in Enterobacteriaceae. These carbapenemases are principally metallo-enzymes that possess a zinc ion in their active site. Enterobacter cloacae strain NOR-1, is resistant to imipenem by production of an inducible chromosomal beta-lactamase which is not a metallo enzyme but an Ambler class A beta-lactamase. This enzyme displays a strong hydrolytic activity against carbapenems but also against cephamycins which has never been previously observed for beta-lactamases of this class. These properties suggest that NMC-A possesses an original tridimensional structure of its active site allowing hydrolysis of molecules generally protected by the configuration of their lateral chain. Crystallographic study of this enzyme will permit to understand its mechanism of action and serve the development of new beta-lactams stable to b-lactamase hydrolytic activity. PMID- 9872011 TI - [Synthesis of multifunctional surfactants for elaboration of micellar model systems of enzymes]. AB - In order to provide a closer analogy between micelles and enzymes, the design of functionalized micellar systems have been undertaken. This paper presents the synthesis of surfactant cationic molecules which contains either a free or a protected aldehyde group. An ammonium quaternary surfactant with two functional groups, aldehyde dimethyl acetal and imidazole, has also been synthesized. PMID- 9872012 TI - [New approaches for encapsulation of peptides into poly(lactic/glycolic acid) microspheres]. AB - The aim of the work was to develop small microspheres made from a biodegradable polymer, poly(lactide-co-glycolide), in order to entrap small peptides. Microspheres prepared by a water-in-oil-in-water emulsion solvent evaporation technique displayed a mean diameter below than 10 microns and showed high encapsulation efficiency of a 33 amino acid peptide (V3 BRU). In vitro release kinetics studies showed that such microparticles could be employed for both oral immunization and controlled release. The encapsulation of a seven aminoacid peptide in the same conditions, led to a very low encapsulation efficiency. In order to increase the entrapment efficiency, two strategies were adopted: taking into account the solubility of pBC 264 at different pH, a pH gradient was created to prevent the leakage of the encapsulated peptide into the outer aqueous phase. The inner aqueous phase was maintained at basic pH where the peptide was soluble, while the external aqueous phase was acidic: ovalbumin was added during preparation to stabilize the inner emulsion. These two strategies allowed to increase significantly the encapsulation rate of pBC 264. Nevertheless, the in vitro release kinetics of the peptide were strongly influenced by the presence of ovalbumin which seems to form pores in the microsphere structure (80% of the total peptide content was released after 30 minutes). By contrast, when ovalbumin was replaced by Pluronic F 68 microspheres did not have pores, thus the release profile and the extent of the burst were much smaller. When microspheres were stereotactically implanted in the rat brain, in vivo release profiles were in good agreement with the release observed in vitro. In conclusion, these microspheres are well suited for the slow delivery of neuropeptides in the brain, a feature expected to facilitate the study of long term effects of these compounds. PMID- 9872013 TI - [The existence of "Nederwiet", a new fact in the history of cannabis]. AB - Since the beginning of the 1990s, France has been faced with a new problem of drug abuse. More and more people are growing Dutch cannabis, commonly called "nederwiet". Grown mostly indoors, its culture requires very specific and sophisticated techniques and materials. The Dutch have produced a cannabis with a very high percentage of tetrahydrocannabinol. We have conducted a morphological and chemical study of one of the known varieties of nederwiet, "super-skunk". Using gas chromatography combined with mass spectrometry we have identified the cannabinoid components. The question of the Dutch regulations on drug abuse and the discussion on legalizing cannabis use in France raises many unanswered questions of great importance. PMID- 9872014 TI - Flavonoids from leaves of Olea europaea L. cultivars. AB - We isolated and identified the following flavonoid compounds from the dried leaves of some blooming cultivars of Olea europaea L.: hesperidin, rutin, luteolin-7-O-glucoside, apigenin, apigenin-7-O-glucoside, quercetin, kaempferol. The structure of the isolated flavonoids was determined by UV, 1H-NMR, 13C-NMR, HPLC. PMID- 9872015 TI - Laboratory evaluation of molluscicidal activity of extracts from Cotula cinerea (L) and Quercus lusitania var. infectoria galls (Oliv.). AB - In this work, we have studied the molluscicidal activity of different extracts obtained from Cotula cinerea and Quercus lusitania var. infectoria galls. The hydroalcoholic extract of Cotula cinerea, acetonic extract and gallotanin of Quercus infectoria galls have presented high activity against Bulinus truncatus. The hydroalcoholic extract of Cotula cinerea was fractionated by chromatography on silica gel column. We have isolated two very active fractions at concentrations respectively of 52.5 and 27.5 ppm. PMID- 9872016 TI - [The phenomenon of globalization in the pharmaceutical industry]. AB - The globalization of international trading has increased so much over the last ten years that it would seem, from an economical point of view, irreversible. The Pharmaceutical Industry is very much part of this movement and, with the cost of innovation and the necessity to achieve critical mass, the health industry is continuously reorganising on a worldwide scale. A study of the Pharmaceutical Industry over the past 20 years has shown an ever increasing globalization particularly for the most innovative drugs. This global reorganisation of the pharmaceutical environment is far from being finalized, to the contrary it will continue for several years to come. The point is not to agree or disagree but succeed, not only in terms of economic progress but also in terms of resolving social and public health care issues. Medication must be available to all, not only in our own country, but worldwide. Is there a response to this increasing globalization of the economy other than an increasing solidarity locally, regionally and worldwide? PMID- 9872017 TI - [Reflections on traditional Chinese medicine and its pharmacopoeia]. AB - Traditional Chinese medicine considers man at the center of the universe as an antenna between celestial and earthly elements. The world is a single unit resulting from one Fundamental Unit. Its movement gives rise to yin and yang which ar two antitheric aspects. When these two energies fall out of harmony, disease develops. The physician should thus take into account this concept when caring for patients and establish an energy diagnosis. Several possibilities are given at treatment which should reestablish the balance between the yin and the yang. PMID- 9872018 TI - Uterine myoma after cessation of gonadotropin-releasing hormone agonist: ultrasound and histopathologic findings. AB - BACKGROUND: This study attempted to search for the possible mechanism of regrowth of uterine myoma after cessation of gonadotropin-releasing hormone agonist (GnRHa) therapy. METHODS: Five premenopausal women presenting with symptomatic uterine myoma were prospectively studied in this trial. All patients were treated with a subcutaneous injection of goserelin depot 3.6 mg every four weeks for 16 weeks. Clinical examinations as well as hormonal and ultrasound determinations were performed before, during and after treatment to monitor the efficacy of therapy. At the end of the treatment period, all patients underwent myomectomy. The ultrastructural change of the myoma was evaluated postoperatively. RESULTS: The volume of the uterus decreased 21-57% and the volume of the uterine myoma decreased 21-65% after therapy. There was no significant change in resistance index of the uterine vessels or major vessels supplying the uterine myoma between pretreatment and post-treatment values. The morphologic features of the treated myoma showed marked cellular shrinkage and loss of myofibrillar structure. CONCLUSIONS: There is no significant cellular damage to the cellular structure that contributes to regrowth of treated intramural uterine myoma following cessation of treatment. PMID- 9872019 TI - Medial synovial entrapment syndrome of the hip in Chinese children. AB - BACKGROUND: "Medial synovial entrapment syndrome of the hip" is a preliminary name given by the authors to describe the topic of this paper. The authors hope to elicit the concern of clinicians and further explore the feasibility of this nomenclature for the described syndrome. METHODS: A total of 26 children with synovial disorder of the hip complained of limited motion and pain of various degrees in the hip, and a limp or refusal to bear weight. Clinical presentation, image studies, course and management were thoroughly reviewed and compared with other well-established synovial disorders of the hip. RESULTS: Twenty-two of the patients were cured by bedrest, skin traction and manipulative reduction. The remaining four patients were treated with surgery and had satisfactory results. Because of its unique clinical manifestation and the good results obtained from manipulative reduction, it is difficult to compare this condition with any other well-established disease entities. Thus, the authors suggest medial synovial entrapment syndrome of the hip (SES) as a preliminary name. CONCLUSIONS: Based on the 26 Chinese children with SES studied, the authors used clinical, anatomic and biomechanical bases for deriving this nomenclature. The similarity between transient synovitis and SES is discussed. PMID- 9872020 TI - Genetic analysis of the insulin receptor gene in Chinese patients with extreme insulin resistance. AB - BACKGROUND: Many patients with extreme insulin resistance in combination with acanthosis nigricans are reported to have defective insulin receptor genes. Because acanthosis nigricans is commonly accompanied by severe hyperinsulinemia and obesity, and obesity is a major factor in insulin resistance, this study was initiated to assess the prevalence of mutations in the insulin receptor gene in Chinese patients with extreme insulin resistance defined by hyperinsulinemia, obesity and acanthosis nigricans. METHODS: Exons 1-22 of the insulin receptor gene were amplified by polymerase chain reaction (PCR) by from genomic DNA from 13 young subjects with clinical and metabolic features of extreme insulin resistance. They were also screened for nucleotide variation using single-strand conformation polymorphism (SSCP) combined with nucleotide sequence analysis. RESULTS: Variant SSCP patterns were detected in exons 1, 3, 6, 11, 12 and 17 of the insulin receptor gene. However, sequencing of amplified DNA fragments revealed that none of the variations were mutations. The SSCP variant in exon 1 was caused by a novel intron polymorphism (G-->T at position 13, 5' intron), while the SSCP variant in exon 12 was caused by a novel silent polymorphism Thr789 (ACG-->ACA). Variants in exons 3 and 17 corresponded to known silent polymorphisms: Gln276 (CAA-->CAG) and His1068 (CAC-->CAT), respectively. Another three variants in exons 3, 6 and 11 were also identified as known polymorphisms in the flanking introns. In addition, two alterations, a silent polymorphism Gly8 (GGA-->GGG) and a polymorphism in the 3' flanking intron (T-->G at position 74), were observed in exon 1 of the insulin receptor gene. CONCLUSIONS: Although the prevalence of insulin receptor gene mutations in this Chinese study population might be underestimated because of the sensitivity of SSCP, these results suggest that mutations at the insulin receptor locus are uncommon in subjects with features of hyperinsulinemia, obesity and acanthosis nigricans. PMID- 9872021 TI - Effects of acute brainstem compression on auditory brainstem response in the guinea pig. AB - BACKGROUND: The purpose of this study was to establish the norm for parameters of auditory brainstem response (ABR) in the guinea pig and to investigate if acute brainstem compression results in significant changes to these parameters. METHODS: Thirty-six guinea pigs with positive Preyer's reflex were anesthetized. A craniectomy was performed to remove the right occipital bone and the dura mater was opened to expose the brain, cerebellum and cerebellopontine angle (CPA). A small inflatable balloon was placed into the CPA precisely and slowly. ABR was recorded before incision of the skin as a baseline value, after placement and after inflation of the balloon with water at 0.1-ml intervals. RESULTS: Five stable peaks were recorded in 27 experimental animals. When the balloon was inflated with 0.1 ml water, the absolute latency (AL) of peaks IV and V and the interpeak latency (IPL) of peaks III and IV, and IV and V were prolonged. The amplitude ratios (AR) of peaks II, III, IV and V to peak I decreased. Inflation of the balloon with 0.2 ml of water caused further elongation of ALs of peaks IV and V and decreases in each AR. When the balloon volume increased to 0.3 ml, peak V became unrecognizable and peaks III and IV showed significant elongation of AL; peaks I and II did not show significant change in ALs. Further increase of the balloon volume to 0.4 ml resulted in disappearance of peaks III, IV and V; AL of peak II was also elongated. However, the amplitude and AL of peak I remained unchanged. Similar changes were observed in IPLs. CONCLUSIONS: This study establishes the norm of parameters of ABR in guinea pigs and demonstrates that acute brainstem compression causes elongation of ALs and IPLs of peaks II, III, IV and V. This suggests that peaks II, III, IV and V come from the brainstem and that peak I is not generated from the brainstem in the guinea pig. PMID- 9872022 TI - Complicated urinary tract infection: analysis of 179 patients. AB - BACKGROUND: The aim of our study was to investigate the incidence, bacteriology, management and outcome of complicated urinary tract infections (UTIs) at the Veterans General Hospital-Taipei. METHODS: Between June, 1993, and July, 1994, medical records of 2,566 patients admitted to the Division of Urology, Veterans General Hospital-Taipei, were retrospectively reviewed. Of these patient, 1,322 had a diagnosis of benign prostatic hyperplasia (BPH), 607 were admitted for renal stones, 496 for ureteral stones, 75 for transitional cell carcinoma (TCC) of the urinary bladder, 47 for renal tumors and 19 for TCC of the ureter. Among all patients studied, 179 (6.98%) acquired a complicated UTI. Of these, 81 were admitted for BPH, 46 for renal stones, 42 for ureteral stones, five for TCC of the urinary bladder, three for renal tumors and two for TCC of the ureter. RESULTS: Of the 179 patients with complicated UTIs, 155 were men and 24 were women. The urine culture positive rate was 76.0% (136/179) and the most common bacteria were Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa. The principle mode of treatment included parenteral antibiotics and urinary diversion (percutaneous nephrostomy and Foley catheterization), when necessary. The infection control rate for these complicated UTIs was 96.3% for BPH, 95.5% for renal stone, 97.6% for ureteral stone, 80% for TCC of the urinary bladder, 100% for renal tumor and 100% for TCC of the ureter. Mortality due to complicated UTI was 3.9% (7/179). CONCLUSIONS: We concluded that the prognosis of complicated UTI is good if diagnosis and appropriate treatment are given promptly. Early drainage to relieve obstruction and intravenous antibiotics are initially necessary. Surgical intervention is required to resolve functional or structural abnormalities after the UTI has been controlled. PMID- 9872023 TI - Color Doppler ultrasound velocimetry of internal carotid arteries in normal pregnancies. AB - BACKGROUND: There are few reports on maternal cerebral circulation during pregnancy. Using the hypothesis that cerebral blood flow resistance decreases progressively with advancing gestational age (GA) as a consequence of estrogenic effects on the vascular system during pregnancy, we calculated the pulsatility index (PI) and the mean velocity (MV) of the maternal right internal carotid artery (ICA) in order to build fitted reference centiles. METHODS: A total of 315 pregnant women with a single fetus were studied at 20-42 weeks' gestation. The patients had uncomplicated singleton pregnancies and were without any known fetal or maternal risk factors. Duplex color ultrasound with a pulsed Doppler scanner (7.5 MHz) was used to determine the PI and MV of the maternal ICA. RESULTS: Among the 315 healthy pregnant women, the PI of maternal ICA decreased progressively with advancing GA, PI50th = Exp (0.3124-0.0084 x GA), (p = 0.0003), during the second half of pregnancy. The predicted 50th percentile PI values of the ICA decreased from 1.117 (5th% = 0.755, 95th% = 1.654) at 24 weeks' gestation to 0.917 (5th% = 0.659, 95th% = 1.448) at 40 weeks' gestation. The MV of the maternal ICA also decreased progressively with advancing GA, MV50th = Exp (3.855 0.0093 x GA), (p = 0.0005), during the second half of pregnancy. The predicted 50th percentile MV values in the ICA decreased from 37.811 cm/sec (5th% = 24.177 cm/sec, 95th% = 59.138 cm/sec) at 24 weeks' gestation to 32.591 cm/sec (5th% = 20.818 cm/sec, 95th% = 51.021 cm/sec) at 40 weeks' gestation. CONCLUSIONS: Both the PI and MV of the maternal ICA decreased with GA during the second half of normal pregnancy as a result of intracerebral vessel dilatation. PMID- 9872024 TI - Os supratrochlear dorsale of the elbow: a report of two cases. AB - The os supratrochlear dorsale, an accessory ossicle of the elbow, has rarely been described in the literature. Rather than being caused by trauma, it is subject to trauma by elbow extension with secondary chondrometaplasia, resulting symptomatology. The aim of this report is to remind readers that its presence is crucial for both diagnostic and therapeutic purposes. Arthroscopic removal may be required to relieve persistent pain, catching and limited range of motion. PMID- 9872025 TI - Intraluminal mucin pool in mucinous gastric adenocarcinoma: a case report. AB - Mucinous gastric carcinoma is a rare pathologic subtype of gastric adenocarcinoma. Whether the mucin behaves aggressively as in mucinous colorectal carcinoma is still controversial. Most mucinous gastric carcinomas are diagnosed from surgical specimens. The mucinous gastric carcinoma in this case report was discovered preoperatively according to its characteristic presentation. An upper gastrointestinal endoscopic examination showed a round protruding tumor of greater than 4 cm in size on the mid-body of the stomach; it had an uneven, friable and shiny surface. The surface was coated with a sticky layer of mucin like substance, which persisted even after the aspiration of the gastric juice. Endoscopic ultrasonography (EUS) revealed a large heterogeneous hyperechoic tumor mass, originating from the mucosal and submucosal layers, on the body of the stomach. The mass was covered with a thick layer of hypoechoic amorphous substance. Hence, a mucin-producing tumor was suspected. Subsequent surgical biopsy proved the mass to be a moderately differentiated mucinous adenocarcinoma. This case illustrates the first endoscopic ultrasonographic report of an intraluminal mucin pool as a hypoechoic substance, which is quite different from the hyperechoic presentation of intramural mucin lakes. In preoperative evaluation, EUS is not only important for determining the depth of tumor invasion, but it is also useful in differentiating mucinous gastric carcinoma from nonmucinous gastric carcinoma. PMID- 9872026 TI - Flutamide-induced liver injury: a case report. AB - Flutamide is a nonsteroidal antiandrogen agent. Since it was marketed in February of 1989 in the USA for treatment of prostate cancer, its potential for hepatotoxicity has been reported in Western countries. Here we report the case of a 72-year-old patient who suffered from general malaise, poor appetite, nausea and jaundice after six months of flutamide therapy for the treatment of prostate cancer. He had no past history of liver disease and was not receiving other medications. Liver biochemistries revealed elevated serum alanine aminotransferase and aspartate aminotransferase concentrations of up to 1,035 U/l and 745 U/l, respectively. Serum total bilirubin concentration was elevated to 7.0 mg/dl. Serologic markers for acute viral hepatitis were all negative. Serum antinuclear antibody, antimitochondrial antibody and antismooth-muscle antibody were also negative. Percutaneous liver biopsy revealed pericentral zonal necrosis with bridging hepatic necrosis. The patient's clinical symptoms and signs began to improve after discontinuation of flutamide, and his liver function had returned to normal three months later. Roussel Uclaf causality assessment for adverse drug reaction confirmed the diagnosis of drug-induced liver injury. This case reminds us that patients who are receiving flutamide should be regularly monitored for liver function. If drug-induced liver injury is suspected, flutamide must be discontinued promptly to avoid progression of liver injury. PMID- 9872027 TI - Successful treatment of a bulky seminoma in an abdominal undescended testis: a case report. AB - An undescended testis is a major risk factor for the development of testicular malignancy. Herein we report the case of a 40-year-old man with a bulky seminoma arising in an uncorrected abdominal undescended testis. The definitive diagnosis was established by results of sonoguided biopsy and tumor marker studies. The patient was successfully treated with initial cisplatin and etoposide combination chemotherapy followed by resection of the residual tumor. The optimal diagnosis and treatment of this rare disease is discussed. PMID- 9872028 TI - One-week low-dose triple therapy is effective in treating Helicobacter pylori infected patients with bleeding peptic ulcers. AB - Proton pump inhibitor (PPI)-based triple therapy, which combines a PPI and two antibiotics, is highly effective in eradicating Helicobacter pylori infection in peptic ulcer patients, even if given for only 1 week. However, the application of this regimen in patients with bleeding ulcers has not been adequately investigated. We studied the effectiveness of triple therapy in treating 122 patients with proven H. pylori infection, and bleeding stigmata on endoscopy; 97 had duodenal ulcer (DU), 15 had gastric ulcer (GU), and 10 had both types of ulcers. A regimen of omeprazole (20 mg), metronidazole (500 mg), and clarithromycin (250 mg) twice daily was administered for 1 week as soon as the patient could eat normally after bleeding, followed by omeprazole (20 mg) daily for 3 additional weeks. Follow-up endoscopy and 13C-urea breath tests (UBTs) were performed at least 4 weeks after triple therapy. A total of 104 patients completed the study, 83 with DU, 12 with GU, and nine with both. The overall ulcer healing rate was 97.1% and the eradication rate was 91.3%. Patients with and without H. pylori eradication did not differ significantly in terms of age, gender, UBT titer, units of blood transfused, or interval between endoscopy and the beginning of triple therapy. We conclude that 1-week low-dose PPI-based triple therapy is effective in eradicating H. pylori infection in patients with bleeding peptic ulcers. When followed by 3 weeks of additional PPI treatment, a satisfactory ulcer healing rate can also be achieved. PMID- 9872029 TI - Polymorphisms of twelve short tandem repeat loci in a Taiwanese population and their application in parentage testing. AB - With the advancement of techniques in molecular biology, rapid, sensitive, and reliable methods of DNA typing for parentage testing have become available. In this study, we evaluated the usefulness of multiplex polymerase chain reaction (PCR) with 12 unlinked short tandem repeat (STR) loci for paternity testing in Taiwan. The genetic informativeness of this test was then compared with that of conventional human leukocyte antigen (HLA) analysis in 167 parentage studies. The 12 STR loci alone provided a cumulative power of exclusion of up to 0.9998. Paternity was excluded in 59 (35.3%) cases, including 40 of 112 paternity trios and 19 of 55 paternity duos. In the 40 trios in which paternity was excluded, a mean of 6 (range, 3-9) incompatible STR markers were in the 19 duos in which paternity was excluded, a mean of 4 (range, 1-8) incompatible STR markers were noted. In the 72 trios in which the alleged paternity could not be excluded, the mean probabilities of paternity (PP) were 90.6863% with HLA testing alone, 99.9847% with STR analysis alone, and 99.9972% with combined HLA and STR analysis. In the 36 duos in which the alleged paternity could not be excluded, the mean PPs were 81.4768% with HLA testing alone, 99.6124% with STR analysis alone, and 99.9145% with combined HLA and STR analysis. These results suggest that STR analysis is very powerful when used alone for paternity trio testing and when combined with conventional serologic HLA typing for duo parentage testing in the Taiwan population. PMID- 9872030 TI - Clinical characteristics and coronary risk factors of patients with low concentrations of serum low-density lipoprotein cholesterol and total cholesterol. AB - We investigated the clinical characteristics and coronary risk factors of Chinese patients with suspected coronary artery disease (CAD) having low serum concentrations of both low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC). Of 1,450 patients with suspected CAD (age range, 30-92 years; 948 men and 502 women), 760 had established CAD. The patients were divided into three groups according to lipid profile patterns. Group 1 patients (n = 138) had low LDL-C concentrations (< 100 mg/dL) and low TC concentrations (< 160 mg/dL). They were characterized by lower triglyceride concentrations, lower frequencies of high TC/high-density lipoprotein cholesterol (HDL-C) ratios (> 5) and LDL C/HDL-C ratios (> 5), and lower frequencies of a family history of CAD and obesity. Group 3 patients (n = 610) had LDL-C concentrations of 130 mg/dL or above and TC concentrations of 200 mg/dL or above, much higher than in group 1. The prevalence of CAD was 41.3% (57/138) in group 1. 46.7% (328/702) in group 2, and 61.5% (375/610) in group 3. Groups with higher TC and LDL-C concentrations had a higher CAD prevalence. Coronary risk factors of group 1 patients appeared to be low HDL-C concentration, high TC/HDL-C ratio, advanced age, cigarette smoking, hypertension, and diabetes mellitus. Among these risk factors, HDL-C and hypertension were independent predictors of CAD. Unlike in the other two groups, hypertension was the only independent nonlipid risk factor. We conclude that in therapy or prevention of CAD, the goals should be to reduce LDL-C concentration to below 100 mg/dL and the TC concentration to below 160 mg/dL. However, other risk factors should also be considered. PMID- 9872031 TI - Effects of zinc deficiency on endogenous antioxidant enzymes and lipid peroxidation in glomerular cells of normal and five-sixths nephrectomized rats. AB - We evaluated the effects of zinc deficiency on the activities of endogenous antioxidant enzymes and lipid peroxidation in rat glomerular cells (GCs). Male Sprague-Dawley rats (n = 48) were fed a zinc-deficient diet and deionized distilled water for 1 week to induce zinc deficiency. Half of the rats (zinc deficient group) continued on this diet for 4 weeks, and the other half (zinc replete group) were maintained on the same diet but with zinc-supplemented water (150 mg/Lzinc sulfate solution). Half of each group underwent five-sixths nephrectomy, while the other half underwent a sham operation. Another 12 normal rats (controls) were fed standard rat chow (containing 23.4% protein and 70 ppm zinc) and drank deionized distilled water. The zinc-deficient rats, including sham and five-sixths nephrectomized rats, showed severe growth retardation and poor appetite. Their mean plasma zinc concentrations were half that of normal control rats, but their plasma copper concentration was significantly higher than that of the control rats. Zinc supplementation corrected the abnormality of plasma zinc and copper concentrations and the loss of body weight in zinc deficient rats. Zinc-deficient rats exhibited lower renal creatinine clearance and higher GC-malondialdehyde (GC-MDA) than zinc-replete rats. The remnant kidney of all five-sixths nephrectomized rats, including zinc-deficient and zinc-replete rats, showed a compensatory elevation in renal creatinine clearance and increased GC-MDA concentrations. Zinc concentrations in the renal cortex were decreased in zinc-deficient rats and the activities of GC-superoxide dismutase and GC glutathione peroxidase were increased, while zinc-replete rats exhibited normal activities of GC-superoxide dismutase and GC-glutathione peroxidase. We suggest that zinc deficiency enhances the formation of reactive oxygen species but does not affect the activities of endogenous antioxidant enzymes in glomerular cells. PMID- 9872032 TI - Factors related to delayed treatment and posttreatment symptom severity in Taiwanese patients with benign prostatic hyperplasia. AB - We evaluated the sociodemographic and clinical factors of delayed treatment and posttreatment symptom severity in outpatients with benign prostatic hyperplasia (BPH). The study included 146 BPH patients treated at the National Taiwan University Hospital in early 1997. All patients were treated with alpha adrenergic antagonists or finasteride for at least 2 weeks. A questionnaire based on Andersen's Health Behavior Model was used to assess various sociodemographic features, while the pre- and posttreatment symptoms severity was rated according to the International Prostate Symptom Score (IPSS). Multiple logistic regression was used to assess the associations of these factors with delayed treatment and posttreatment symptom severity. Subjects who had recently quit smoking or were blue-collar workers tended to delay treatment, while those who chose a medical center as the care provider for chronic diseases tended to be less likely to delay treatment. However, none of these associations were statistically significant. No enabling factors (income, insurance) or need factors (symptom scores) evaluated were associated with delayed treatment. Predisposing factors associated with higher posttreatment symptom severity were delayed treatment (over 12 months) (adjusted odds ratio [OR]: 2.67, 95% confidence interval [CI]: 1.16-6.16), quitting smoking (adjusted OR: 4.47, 95% CI: 1.34-14.94), and having never smoked (adjusted OR: 3.73, 95% CI: 1.15-12.11). Subjects with severe pretreatment symptoms were far more likely than subjects with mild pretreatment symptoms to have severe symptoms after treatment (adjusted OR: 52.69, 95% CI: 54.46-621.90). Our findings, though based on a limited number of subjects, suggest sociodemographic factors rather than objective clinical attributes (prostate specific antigen level, prostate volume, and urodynamic results) are associated with delayed treatment in Taiwanese men with BPH. Both pretreatment symptom severity and sociodemographic factors are related to posttreatment symptom severity. PMID- 9872033 TI - Treatment of developmental dysplasia of the hip after failed open reduction. AB - It is difficult to obtain a good result by secondary open reduction if a primary open reduction for developmental dysplasia of the hip (DDH) fails. Complications such as avascular necrosis of the femoral head and subluxation of the hip are common. In this study, we retrospectively reviewed the causes of failure of primary open reduction and the final clinical and radiographic outcomes of 32 patients (34 hips) with DDH who underwent repeat open reduction and other procedures from January 1982 to December 1995. The ages of the patients at the time of the secondary operation ranged from 1.5 to 16.5 years (mean, 5.9 yr). The interval from the primary open reduction to the secondary procedure ranged from 3 days to 10 years (mean, 8.9 mo). In most cases (30 hips), the position of the redislocated femoral head was Tonnis grade 3 or 4. Avascular necrosis of the femoral head was evident in about half of the hips before the secondary open reduction. The most common cause of failure of the primary operation was a tight inferior capsule and transverse acetabular ligament, which blocked complete reduction. At a mean follow-up period of 42 months (range, 24-147 mo) after the secondary operation, the radiographic classification was Severin class 1 or 2 in 15 of the 34 hips, and Severin class 3 or worse in the remaining 19 hips. Clinically, according to the modified McKay criteria, 18 of the 32 patients (18 hips) had excellent or good results, and three patients (four hips) had poor results. In conclusion, the main cause of failure of the primary open reduction of DDH was technical error. We believe that detailed preoperative evaluation is critical for the success of primary open reduction of DDH. PMID- 9872034 TI - Compliance with the law and appropriate medical standards during interhospital transfers. AB - Interhospital transfer patients constitute a significant proportion of medical center emergency department (ED) patients in Taiwan. Many such transfers are poorly planned and put the patient at risk. We wished to evaluate the safety and compliance with the Taiwan Medical Law among patients transferred to the Linkou Chang Gung Memorial Hospital ED from other health care centers. We performed a prospective, cross-sectional, observational study on 1,056 patients transferred from August 15 to September 30, 1997. Of these patients, 357 were critically ill or injured and only 160 received adequate pretransfer stabilization. The major omissions included: 1) failure to intubate in 121 (55%) of the 220 patients in severe respiratory distress or unprotected patent airways; 2) no intravenous line in 74 (20.7%) of the 357; and 3) inadequate IV lines in 36 (63.2%) of the 57 severely hypotensive patients. Overall, 894 patients were sent with transfer notes, but few indicated whether the referral was to the ED or outpatient department. This added an unnecessary burden for patients with stable longstanding problems who claimed they had been referred to the ED. While the majority of patients (49.4%) were transferred at the request of physicians for further treatment, 28% of the critically ill patients were transferred because of family requests. Physicians accompanied these patients only on seven occasions and nurses on 84 occasions. Despite the 1993 Department of Health policy of pretransfer phone contact with the receiving hospital for critically ill patients, such contact occurred only 10.6% of the time. While the Taiwan emergency medical system, Emergency Medicine, and Critical Care Medicine are all in their developmental stages, a medical and legal noncompliance rate of above 55% for critically ill transfer patients is unacceptably high. The appropriate medical societies and the Department of Health should work in concert to upgrade existing transfer practices. PMID- 9872035 TI - Central retinal vein occlusion associated with cryofibrinogenemia in a young adult--study of skin dynamic capillaroscopy. AB - A 26-year-old woman presented with a 2-month history of intermittent dim vision in her right eye and a diagnosis of central retinal vein occlusion (CRVO) was made. Investigations revealed Raynaud's phenomenon and cryofibrinogenemia. Skin capillaroscopy revealed tortuous vessels and decreased capillary blood-cell velocity in the capillaries of her finger nail fold which coincided with the CRVO attack. After treatment with oral aspirin for 3 months, the CRVO subsided, the capillary blood-cell velocity in the nail fold improved, and serum cryofibrinogen was undetectable. The time course of the cryofibrinogenemia and skin capillaroscopic findings correlated well with her ocular presentations. Study of cryofibrinogen should be included in the evaluation of young adults with CRVO. PMID- 9872036 TI - Systemic Penicillium marneffei infection in a child with common variable immunodeficiency. AB - Penicillium marneffei is rarely pathogenic in humans. Most previously reported cases of P. marneffei infection were from Southeast Asia where patients were usually in an immunocompromised state due to human immunodeficiency virus (HIV) infection. The majority of the patients reported in Western countries were immunocompromised by malignancy, especially Hodgkin's lymphoma. In Taiwan, the first case of P. marneffei infection was reported in 1994 and involved an adult with HIV infection. We report a case of systemic P. marneffei infection in a child with common variable immunodeficiency (CVID). The patient, a 4-year, 5 month-old boy, had a 1-year history of oligoarthritis resembling juvenile rheumatoid arthritis (JRA). He developed a low grade fever (38 degrees C) and hepatosplenomegaly 1 month before admission to the hospital. Although cultures of synovial fluid obtained at the time of onset of oligoarthritis did not grow any organisms, cultures of blood, bone marrow, synovial fluid, and lymph node biopsy samples taken during this admission were positive for P. marneffei. Further immunologic studies revealed a profile characteristic of CVID. The fungal infection was finally eradicated by combined therapy with amphotericin B, fluconazole, itraconazole, and regular immunoglobulin replacement. This case reminds us that JRA or JRA-like arthritis should be differentiated from septic arthritis caused by rare pathogens in immunocompromised patients. PMID- 9872037 TI - Hypothalamic juvenile pilocytic astrocytoma presenting with intracerebral hemorrhage. AB - Hemorrhage of juvenile pilocytic astrocytomas is very rare. However, it is clinically important because a high probability of a fatal outcome exists if hemorrhage occurs outside the optic nerve. Hemorrhage of a hypothalamic juvenile pilocytic astrocytoma has been reported only once, from an autopsy examination. We present a successfully managed case. The patient, a 34-year-old man, presented with sudden loss of consciousness. Computed tomography (CT) and magnetic resonance imaging demonstrated an exophytic hypothalamic tumor with a hemorrhage. Craniotomy with total removal of the tumor and the hematoma was performed. Histopathologic examination revealed that the tumor was a juvenile pilocytic astrocytoma. Brain CT examination 18 months later revealed no evidence of tumor recurrence. The patient was well, without neurologic deficit, and had a normal social life as of the last follow-up examination 22 months after craniotomy. PMID- 9872038 TI - Histomorphometric evaluation of mucin content in stone-containing intrahepatic bile ducts. AB - Although histologic studies of mucin distribution in the peribiliary glands have been conducted, a quantitative study of mucin content in intrahepatic bile ducts has yet to be reported. In an attempt to evaluate the mucin content in stone containing intrahepatic bile ducts, we conducted a study on 25 surgically resected livers with hepatolithiasis. Specimens from 10 livers without stones served as controls. All specimens were fixed in 10% formalin and sectioned for periodic acid Schiffalcian blue double-stain (PAS-AB; pH 2.5) to evaluate the epithelial mucin content of the intrahepatic bile ducts. The PAS-AB positive area and the total epithelial area were measured with a computerized image analyzer and the PAS-AB index was calculated as the proportion of the PAS-AB positive to the total epithelial area. The histochemical study showed that epithelial cells in both the intramural glands and extramural glands of stone-containing intrahepatic bile ducts stained heavily and homogeneously with PAS-AB, while those of controls stained weakly. The PAS-AB indexes in stone-containing intrahepatic bile ducts were 51.8 +/- 15.88% for mucous cells of intramural glands, 52.86 +/- 9.85% for mucous cells of extramural glands, and 77.29 +/- 21.59% for serous cells of extramural glands. These values were all significantly higher than those of control specimens. However, the PAS-AB index of the epithelial lining in both hepatolithiasis and control specimens were similarly low, indicating the epithelial lining does not secrete much mucous glycoprotein. The results of this study led us to conclude that stone-containing intrahepatic bile ducts contain an abundant amount of mucous glycoprotein, and mucin is secreted from the peribiliary glands, not from the epithelial lining of the bile ducts. PMID- 9872039 TI - [(In-)dependence of editors of medical periodicals]. PMID- 9872040 TI - [Asthma therapy in children and adults. Recommendations of the German Respiratory League of the German Society of Pneumology]. PMID- 9872041 TI - [Continuous blood glucose monitoring: the acute effect of acarbose on blood glucose variations]. AB - BACKGROUND: Acarbose, a pseudo-tetrasaccharid, inhibits intestinal alpha glucosidases, effects a reduction of postprandial hyperglycemia and is particularly used in the treatment of patients with type-2 diabetes mellitus. The aim of the study is to analyse by a continuous blood glucose measurement the acute effect of acarbose after a carbohydrate loading and during a 12 hours period. PATIENTS AND METHODS: We examined 10 patients with type-2 diabetes mellitus (mean age 59.2 +/- 3.79, HbA1 9.2 +/- 0.26%) treated with sulfonylureas and/or insulin after a carbohydrate meal and 12 hours during daytime, to test whether the first application of acarbose influences the mean blood glucose or the blood glucose amplitudes. Four measurements were enrolled using a portable continuous blood glucose sensor (Glucosensor, unitec Ulm). A measurement after a carbohydrate loading (Fresubin, 500 kcal, 69 g carbohydrate) with 100 mg acarbose (Glucobay) was followed by a 12-hour measurement during daytime with 3 x 100 mg acarbose and standard diet. These measurements were repeated without acarbose. RESULTS: After a carbohydrate loading, the mean blood glucose level (AUC 44,320 +/- 10,660 with acarbose vs. 61,390 +/- 12,590 without acarbose; mean + SD; p = 0.004) decreased by 28%. During daytime blood glucose levels were not significantly decreased (165.7 +/- 50.3 mg/dl vs 183.7 + 67.4 mg/dl; p = 0.1) although the postprandial blood glucose amplitudes after the 3 meals were reduced significantly (85.90 +/- 24.6 mg/dl vs 106.5 +/- 20.5 mg/dl; p = 0.02). CONCLUSIONS: Continuous blood glucose monitoring indicated that acarbose diminished mean blood glucose levels after a carbohydrate loading in patients with type-2 diabetes mellitus, but not during 12 hours of standard diet, although blood glucose amplitudes decreased. Long-term improvements of metabolism by acarbose may therefore be related to the reduction of blood glucose amplitudes which is likely to reduce toxic effects of glucose on islet cell function. PMID- 9872042 TI - [Antibiotic sensitivity of important pathogens of bacterial respiratory tract infections in Northeast Germany]. AB - BACKGROUND: There is still a lack of comprehensive study results about resistance of bacterial respiratory pathogens from the east of the Federal Republic of Germany. METHODS: In Greifswald we isolated in 1995 and 1996 320 strains of typical pathogens and tested their susceptibility to 14 antibiotics, using the microbouillon dilution method. RESULTS: Pneumococci and beta-haemolytic streptococci were completely susceptible to penicillin and all other beta-lactam antibiotics, clindamycin and vancomycin. 4.8% of pneumococci and 10.0% of Streptococcus pyogenes isolates were resistant to erythromycin. This should lead to a more differentiated use of macrolides as concerns these pathogens, especially as penicillin still has high efficacy in these cases. Pneumococci had resistance rates of 4.8%, 9.5% and 6.7% to cotrimoxazole, tetracycline and ofloxacin, respectively. No strain of Haemophilus influenzae produced a beta lactamase, 2.6% of strains were relatively resistant to ampicillin, even in combination with sulbactam. Cefaclor, erythromycin and tetracycline had restricted efficacy of 16.7%, 14.1% and 53.8%, respectively. Cefuroxime, cefixime, cefepime, imipenem and ofloxacin all had complete efficacy. 77.8% of Moraxella catarrhalis isolates were beta-lactamase positive. Strains were susceptible to erythromycin, ofloxacin and all tested beta-lactam-antibiotics except ampicillin and cefaclor. There was a relative resistance to clindamycin and tetracyclin of 22.2% and 2.2%, respectively. DISCUSSION: We present our results in national and international comparison, describe tendencies of resistance, give reasons for the low incidence of penicillin resistance in pneumococci in Germany and draw conclusions for empirical chemotherapy. PMID- 9872043 TI - [Non-thyroid illness" or changed thyroid hormone parameter syndrome with non thyroid illnesses]. AB - BACKGROUND: The multiple effects of systemic illness on thyroid economy are commonly referred to "non-thyroidal illness" (NTI) or "sick euthyroid syndrome". The various aspects of this common syndrome are summarized in this article. STUDIES: Results of the relevant studies published during the past 25 years were evaluated. The influence of the underlying illness and of drug administration was especially emphasized. RESULTS: The most common abnormalities in NTI are 1. the "low-T3 syndrome" due to a decreased T3 generation from T4 by a reduced activity of 5'-deiodinase (a selenoprotein); 2. the "low-T3 low-T4 state", which is associated with a poor prognosis. The low T4-levels are related to a binding inhibitor that displaces T4 from its binding proteins. However, there exists some controversy regarding the character of this binding inhibitor. 3. The high-T4 state is often found in acute psychiatric and liver diseases. The nutritional status of the patients and drugs known to influence thyroid hormone parameters have to be considered when patients with NTI are evaluated. Some difficulties may arise, when there is evidence of coexisting thyroid disease. Here aside from further biochemical evaluation such as thyroid antibodies, thyroid ultrasound and a thyroid scan have to be performed. CONCLUSION: NTI is associated with various alterations in thyroid hormone parameters when no intrinsic thyroid hormone disease exists. The severity of NTI reflects clinical outcome and clinical amelioration is associated with normalization of thyroid hormone parameters. There is no need for specific therapeutic intervention such as the administration of thyroid hormones in patients with the various forms of the NTI-syndrome. PMID- 9872044 TI - [Duplex ultrasound of the liver and portal vein system]. AB - Duplex ultrasound allows direct access to portal venous and hepatic vein hemodynamics. This method has been accepted as a reliable procedure to detect portal and splenic venous thrombosis, Budd-Chiari's syndrome, portosystemic collateralisation and changes in the portal venous blood flow direction with a high rate of sensitivity. Besides these qualitative findings Doppler flowmetry allows quantitative measurement of portal venous blood flow velocities. The estimation of portal hypertension and the prediction of the risk of variceal bleeding by quantitative Doppler flowmetry is of increasing clinical importance. In monitoring the efficiency of medicational prophylaxis against gastrointestinal bleeding is also helpful. Duplex ultrasound can be used to evaluate the function of portocaval shunts including the intrahepatic stent shunt as well as to detect vascular complications after liver transplantation. Furthermore the method is gaining importance in the diagnosis and monitoring of therapy of acute liver transplant rejection. In case of liver tumors duplex ultrasound is of value for tumor staging and provides information about the dignity on the basis of perfusion patterns. PMID- 9872045 TI - [POEMS syndrome. A rare variant of osteosclerotic multiple myeloma with polyneuropathy, organomegaly, endocrinopathy, M-gradient and skin lesions]. AB - BACKGROUND: A rare multi-organ involvement in plasma-cell dyscrasias has been named POEMS-syndrome: it is a synopsis of monoclonal gammopathy (M-gradient), osteosclerotic bone lesions, peripheral polyneuropathy, organomegaly, endocrinopathy and skin lesions. CASE REPORT: A patient is presented who had a classical manifestation of this disease known mainly in Japan. A monoclonal IgA lambda-gammopathy was determined as cause of a gradually progressive polyneuropathy. The patient had a hypergonadotropic hypogonadism, hyperprolactinaemia, and sclerotic bone lesions. In addition, he showed a changing organomegaly, and hyperpigmentation of the skin. CONCLUSION: As yet, aetiology and pathophysiology are not fully understood. Irradiation or surgical resection of one or several osteosclerotic bone lesions may improve the polyneuropathy or may even lead to a complete remission of all symptoms. Thus, monoclonal immunoglobulins should be searched for in any unclear polyneuropathy, as should be for other symptoms of the POEMS-syndrome. PMID- 9872046 TI - [Addison disease, diagnosis and therapy]. PMID- 9872047 TI - [Ulnar deviation and swan neck deformity of the hands and high titer antinuclear antibodies]. PMID- 9872049 TI - [Detection and evaluation of adverse drug reactions]. AB - Adverse drug reactions (ADRs) occur in about 5% of drug-treated patients. Hospital admissions are caused by ADRs in 5% of patients and roughly 2% of hospitalized patients will experience an ADR. The economic burden of ADRs can only be estimated. Type A reactions can be explained by the pharmacological action of the drugs, and are preventable in many cases. However, Type B reactions involving the immune system and/or idiosyncratic reactions occur rarely and most of them are not fully understood. Genotyping represents an elegant method to explain the presence of abnormal enzyme activities and allows prediction of adverse drug effects in individual cases. Typical time frames have been identified for the occurrence of hypersensitivity reactions, although definite causality assessment is often impeded due to the absence or unavailability of specific laboratory tests and the impossibility of rechallenge. Diagnosis of an ADR is often difficult due to comorbidity and polypharmacy, thus causality assessment is often divergent even between specialists. In Germany, ADRs are reported preferably to the manufacturer of the suspicious drug and then collected and evaluated at the Bundesinstitut fur Arzneimittel und Medizinprodukte, BfArM. However, total number and quality of reported ADRs could be improved. PMID- 9872050 TI - Massage therapy effects. AB - Massage therapy is older than recorded time, and rubbing was the primary form of medicine until the pharmaceutical revolution of the 1940s. Popularized again as part of the alternative medicine movement, massage therapy has recently received empirical support for facilitating growth, reducing pain, increasing alertness, diminishing depression, and enhancing immune function. In this article studies are reviewed that document these effects, and models are proposed for potential underlying mechanisms. PMID- 9872051 TI - Guidelines for the evaluation of dementia and age-related cognitive decline. American Psychological Association Presidential Task Force. AB - Assessment of cognitive functioning among older adults requires specialized training and refined psychometric tools. Psychologists conducting such assessments must learn current diagnostic nomenclature and criteria, gain specialized competence in the selection and use of psychological tests, and understand both the limitations of these tests and the context in which they may be used and interpreted. Assessment of cognitive issues in dementia and age related cognitive decline is a core focus of the specialty of clinical neuropsychology. Therefore, these guidelines are not intended to suggest the development of an independent proficiency. Rather, they are intended to state explicitly some appropriate cautions and concerns for all psychologists who wish to assess cognitive abilities among older adults, particularly when distinguishing between normal and pathological processes. PMID- 9872052 TI - Genetic studies of spectrin: new life for a ghost protein. AB - Spectrin, together with actin and a number of other accessory proteins, forms a submembrane cytoskeletal network in the human erythrocyte ghost. Through an elegant combination of structural, biochemical, and genetic studies, spectrin was shown to be an important determinant of erythrocyte shape and membrane stability. Genetic studies of a novel nonerythroid spectrin (beta H) in Drosophila and Caenorhabditis elegans now reveal that spectrin can influence the shape and stability of whole organisms. Nonerythroid spectrins are proposed to have roles in cell adhesion, establishment of cell polarity, and attachment of other cytoskeletal structures to the plasma membrane. The phenotypes of the beta H spectrin mutations provide an exciting biological context in which to evaluate these roles and perhaps to uncover new ones. PMID- 9872054 TI - Contemporary approaches to protein structure classification. AB - In a similar manner to sequence database searching, it is also possible to compare three-dimensional protein structure. Such methods can be extremely useful because a structural similarity may represent a distant evolutionary relationship that is undetectable by sequence analysis. In this review, we summarise the most popular structure comparison methods, show how they can be used for database searching, and then describe some of the most advanced attempts to develop comprehensive protein structure classifications. With such data, it is possible to identify distant evolutionary relationships, provide libraries of unique folds for structure prediction, estimate the total number of folds that exist, and investigate the preference for certain types of structures over others. PMID- 9872053 TI - Role of the telomeric DNA-binding protein TRF2 in the stability of human chromosome ends. AB - A major issue in telomere research is to understand how the integrity of chromosome ends is preserved. A recent study shows that expression of a dominant negative form of the human telomeric protein TRF2 increases the number of chromosome fusions in immortalized cells and decreases the quantity of G-rich telomeric DNA 3' overhang, the G tail. Consequently, TRF2 appears to control the structure of the very end of the chromosomal DNA molecule and to prevent recombination between two telomeres. Remarkably, the same study reveals a potential role of TRF2 in cell division control. PMID- 9872055 TI - Creating intracellular structural domains: spatial segregation of actin and tropomyosin isoforms in neurons. AB - Actin microfilaments play a direct role in a variety of cell processes. Distinct populations of microfilaments are associated with different cellular compartments, such as growth cones, filipodia, stress fibers, and lamellipodia. It is becoming clear that these different populations are often composed of different isoforms of the two core microfilament components, actin and tropomyosin. This is particularly true in neurons, where actin and tropomyosin isoforms are segregated into different intracellular compartments which correspond to functionally distinct regions of the neuron. Developmental regulation of this isoform sorting suggests a specific role for some isoforms in growth and for others in stabilization of neuronal structure. This provides a mechanism by which a neuron can create and independently regulate intracellular domains composed of microfilaments with different functional properties. PMID- 9872056 TI - Expanding complexity in myotonic dystrophy. AB - Myotonic dystrophy (DM) is a highly variable multisystemic disease belonging to the rather special class of trinucleotide expansion disorders. DM results from dynamic expansion of a perfect (CTG)n repeat situated in a gene-dense region on chromosome 19q. Based on findings in patient materials or cellular and animal models, many mechanisms for the causes and consequences of repeat expansion have been proposed; however, none of them has enjoyed prolonged support. There is now circumstantial evidence that long (CTG)n repeats may affect the expression of any of at least three genes, myotonic dystrophy protein kinase (DMPK), DMR-N9 (gene 59), and a DM-associated homeodomain protein (DMAHP). Furthermore, the new findings suggest that DM is not a simple gene-dosage or gain-or-loss-of-function disorder but that entirely new pathological pathways at the DNA, RNA, or protein level may play a role in its manifestation. PMID- 9872057 TI - The Nck SH2/SH3 adaptor protein: a regulator of multiple intracellular signal transduction events. AB - The process generally termed signal transduction involves the coordinated relay of information from extracellular cues to intracellular effectors, subsequently leading to a specified cellular response. The formation of multimeric protein complexes is a critical step in the activation of most intracellular signal transduction cascades. In many cases, these processes are initiated by a family of molecules consisting of protein association motifs known as src homology 2 and 3 (SH2 and SH3) domains. This review focuses on a group of proteins within this family that lack intrinsic enzymatic functions and consist almost entirely of SH2 and SH3 domains. Termed "adaptors," these proteins serve to physically bridge activated cell surface receptors to various intracellular signal transduction pathways. Here, I briefly summarize current knowledge concerning the various adaptor proteins and place a particular emphasis on Nck. Various data are discussed which collectively support a role for Nck in the regulation of multiple intracellular signaling events. PMID- 9872058 TI - Chromosomal abnormalities and tumor development: from genes to therapeutic mechanisms. AB - This article highlights the recent advances in our understanding of the molecular structure and function of proteins that are activated or created by chromosomal abnormalities and discusses their possible role in tumor development. The molecular characterization of these proteins has revealed that tumor-specific fusion proteins are the consequence of the majority of chromosomal translocations associated with leukemias and solid tumors. A common theme that emerges is that creation of these proteins disrupts the normal development of tumor-specific target cells by blocking apoptosis. These insights identify these chromosomal translocation-associated genes as potential targets for improved cancer therapies. PMID- 9872059 TI - Multidrug resistance mediated by the ATP-binding cassette transporter protein MRP. AB - Resistance to multiple natural product drugs associated with reduced drug accumulation in human tumor cells may be conferred by either the 170 kDa P glycoprotein or the 190 kDa multidrug resistance protein, MRP. Both MRP and P glycoprotein belong to the large and ancient ATP-binding cassette (ABC) superfamily of transport proteins but share only 15% amino acid identify. Unlike P-glycoprotein, MRP actively transports conjugated organic anions such as the cysteinyl leukotriene C4 and glutathione-conjugated aflatoxin B1. Transport of unconjugated chemotherapeutic agents appears to require cotransport of glutathione. MRP and several more recently discovered ABC proteins contain an additional NH2-proximal membrane-spanning domain not found in previously characterized ABC transporters. This domain, whose NH2-terminus is extracytosolic, is essential for MRP-mediated transport activity. This review summarizes current knowledge of the structural and transport characteristics of MRP which suggest that the physiologic functions of this protein could range from a protective role in chemical toxicity and oxidative stress to mediation of inflammatory responses involving cysteinyl leukotrienes. PMID- 9872060 TI - "Painting" the target: how local molecular cues define synaptic relationships. AB - A majority of studies on neuronal growth cones focus on the features that particular groups of neurons share. In contrast, questions such as how specific growth cones respond very differently to the same extrinsic cues require cell specific experimentation. The most succinct cell-specific growth cone responses occur during synaptic targeting. Recent studies have examined one specific growth cone, the Drosophila RP3 motoneuron growth cone, in variously altered microenvironments. In this review, we summarize how such studies are beginning to uncover the repertoire of extrinsic cues that influence the synaptic targeting of a single growth cone. PMID- 9872061 TI - Speculations on the subject of alcohol dehydrogenase and its properties in Drosophila and other flies. AB - The alcohol dehydrogenases (ADH) and their genes (Adh) of Drosophila have been much studied by population and evolutionary biologists. I attempt to put some of these studies into a broad adaptionist perspective, suggesting the co-evolution of this enzyme with the fleshy fruits of angiosperms and fermenting yeasts. I suggest that these events occurred at about the K/T boundary (65 million years ago) and that the typical Drosophila (as exemplified by D. melanogaster) evolved from flies unable to use fermenting substrates as breeding sites. I also hint that the ADH enzymes of other flies (e.g., the tephritid fruit flies) may have evolved independently of those of Drosophila, but from a common ancestral gene. PMID- 9872062 TI - Sperm cells and foreign DNA: a controversial relation. AB - Sperm cells from a variety of species share the spontaneous ability to take up foreign DNA. That feature has been exploited to generate genetically modified animals with variable efficiency in different species. An unexpectedly large set of factors appears to modulate the interaction of sperm cells with exogeneous DNA. The binding is mediated by specific DNA-binding proteins and is antagonized by an inhibitory factor in the seminal fluid. A portion of sperm-bound DNA is internalized in nuclei, a process mediated by CD4 molecules. Sperm interaction with foreign DNA triggers endogenous nuclease(s) that cleaves both the exogenous and the genomic DNA, eventually leading to a cell death process which resembles apoptosis. Internalized foreign DNA sequences reach the nuclear matrix and undergo recombination with chromosomal DNA. From these studies, a surprising network of metabolic functions is beginning to emerge in mature spermatozoa, which are normally repressed and are specifically activated upon exposure to appropriate stimuli. PMID- 9872063 TI - Managing the outcomes of discovered deception. AB - This study analyzed strategies individuals use to manage the discovery of deception in a relationship. A sample of 210 participants completed questionnaires detailing their use of repair strategies subsequent to deception. Findings showed that the use of repair strategies differed across relational types and that the use of prosocial relational repair strategies was positively correlated with the target's increase in trust for the deceiver, the target's expressions of affection for the deceiver, and relational intimacy. Relational satisfaction was positively correlated with the tendency to use prosocial strategies. The deceiver's message selection was partially constrained by perceptions of the target's attributions concerning the relational significance of the deception; prosocial strategies were correlated with the deceiver's belief that the target attributed relational significance to the deceptive act. PMID- 9872064 TI - The importance of employee demographic profiles for understanding experiences of work-family interrole conflicts. AB - This article investigated the reported experiences of work-family interrole conflict differences between American men and women. Participants answered questions on several questionnaires and were classified according to marital status, working or non-working spouse or partner, and presence of children. Overall, men reported significantly higher levels of work-to-family and strain based, family-to-work conflict. Married men with working spouses, with and without children, and married men with children and non-working spouses reported higher levels of time-based and strain-based, family-to-work conflict than women in each corresponding category. The highest time-based, family-to-work conflict was reported by divorced women with children. PMID- 9872065 TI - Intergroup biases and eyewitness testimony. AB - The study examined how the in-group/out-group status of a perpetrator of a distinctly violent crime might influence an eyewitness's evaluation of his behavior and a witness's performance in an identification task. Immigrant and Swedish students saw a film showing a simulated robbery, with an immigrant or a Swede as the perpetrator. Results showed that both groups evaluated an ethnically dissimilar perpetrator as more culpable than an ethnically similar perpetrator. In a line-up task, both immigrant and Swedish participants mistakenly identified an innocent immigrant more often than an innocent Swede. Participants' biased evaluations of the perpetrator are discussed in terms of cognitive and motivational mechanisms. Expectations regarding the typical ethnicity of a perpetrator of this type of crime are suggested to account for the findings of the line-up task. PMID- 9872066 TI - Perception of enhancing self-worth through service: the development of a Service Ethic Scale. AB - A 10-item Service Ethic Scale was developed and its relationship to personal and demographic variables was analyzed using a sample of secondary school administrators. Ninety-two usable surveys were obtained from a sample of 150 secondary school administrators randomly chosen from the 1994-1995 Louisiana Directory of Schools (Bureau of School Accountability, 1994). Results of exploratory factor analysis showed that the scale had 1 factor that explained 63.2% of the variance, and showed excellent reliability. The scale was significantly correlated with endorsement of the Protestant work ethic and the desire to implement an ethical values curriculum. However, the scale was not related to participants' gender, age, job tenure, or school location (i.e., rural or urban). PMID- 9872067 TI - Relation of drinking and eating to masculinity and femininity. AB - The relationship between desirable and undesirable aspects of masculinity and femininity and drinking and eating was investigated. A sample of 144 university women in Australia completed questionnaires that assessed masculinity and femininity, reported drinking, alcohol dependence, eating restraint, frequency of dieting, and overeating. Evidence of a common underlying dimension linking aspects of problem drinking and overeating to undesirable masculine characteristics was found. The results are consistent with the view that women engage in excessive consummatory behaviors such as binging to deal with their gender-role conflict. PMID- 9872068 TI - Self-monitoring and student integration of community college students. AB - This article reports how the Self-Monitoring Scale (M. Snyder, 1974) predicted integration in the student integration model (V. Tinto, 1987, 1993) among a sample of 380 American community college students. A 2 x 2 between-subjects multivariate analysis of variance was performed on 2 dependent variables: academic and social integration. The independent variables were gender and self monitoring (low and high). A significant main effect of self-monitoring was obtained for academic integration. Low self-monitors were significantly more likely to integrate academically than high self-monitors. A significant interaction effect of gender and self-monitoring was obtained for social integration. High self-monitoring men were significantly more likely to achieve social integration than low self-monitoring men. PMID- 9872069 TI - Measuring self-monitoring ability and propensity: a two-dimensional Chinese scale. AB - Self-monitoring characterizes individual differences in a person's ability and propensity to regulate self-presentation (M. Snyder, 1974, 1979). On the basis of this idea and existing scale items (R. D. Lennox & R. N. Wolfe, 1984; M. Snyder, 1974), a 23-item 2-dimensional Chinese Self-Monitoring Scale was devised with subscales measuring the Ability and the Propensity to regulate self-presentation according to situational cues. The two subscales displayed different correlational patterns with the Psychoticism, Extraversion, Neuroticism, and Lie subscales of the Eysenck Personality Questionnaire (EPQ; S. B. G. Eysenck & H. J. Eysenck, 1975). The new scale, with stable factor structure and satisfactory internal consistency, may discriminate among self-presentational styles beyond the prototypes of high and low self-monitoring. PMID- 9872070 TI - Factors affecting behavior toward people with disabilities. AB - Two field experiments were designed to explore ambivalence-induced behavior toward people with disabilities in low-cost helping situations. In Experiment 1, confederates, who either rode in a wheelchair or walked, asked shoppers for a small sum of money and either provided a reason (positive presentation) or did not provide a reason (negative presentation) for the request. Contrary to prediction, more people with a physical disability than without a physical disability were helped, regardless of their presentation. In Experiment 2, the presence of confederates in wheelchairs resulted in significantly fewer violations of parking spaces reserved for people with handicaps. PMID- 9872071 TI - Perceptions of abusive and violent husbands by engaged Arab men in Israel. AB - The predictability of engaged Arab men's perceptions of abusive husbands was tested. The analysis was based on variables reflecting three theoretical perspectives: male dominance, intergenerational social learning, and interpersonal skills deficit. The data were derived from a standardized measurement package completed by 434 Arab men in Israel. The results revealed that a combination of predictors from disparate frameworks best explained the variance in the different criterion variables of the study. Lack of skills for establishing positive communication with one's fiancee, traditional and nonegalitarian expectations of marriage, and experience with or witnessing violence in the family of origin best explained the variance in the belief that abusive husbands should not be held responsible for their behavior. The combination of the first two predictors and negative attitudes toward women best explained the variance in the view of the respondents that abusive husbands should not be punished for their violent behavior. PMID- 9872073 TI - Traumatic events and generational differences in assumptions about a just world. PMID- 9872074 TI - The top 10 MIOSHA violations. PMID- 9872075 TI - Selling or closing your practice. PMID- 9872076 TI - Anticipatory guidance in infant oral health. PMID- 9872077 TI - What you should know about mandatory CDE. PMID- 9872078 TI - Analysis of hyaluronic acid in human gingival crevicular fluid using high performance liquid chromatography. AB - We determined the hyaluronic acid disaccharides, delta Di-HA, in the gingival crevicular fluid (GCF) and whole saliva of patients with periodontal disease, and in the peri-implant sulcus fluid (PISF) from sites around titanium osseointegrated implants, and compared these values with those in the GCF and whole saliva of controls. We also determined values for chondroitin sulfate disaccharide isomers at the same time. Glycosaminoglycans were extracted by digestion with Pronase E, followed by digestion of GAGs with hyaluronidase SD and chondroitinase ACII. Unsaturated disaccharide isomers produced from hyaluronic acid and chondroitin sulfate were analyzed by high-performance liquid chromatography (HPLC). The hyaluronic acid disaccharide delta Di-HA was found in all samples of GCF, PISF and whole saliva. The concentration of delta Di-HA in both GCF and whole saliva of the periodontitis group was greater than that in the controls. There was no difference in the concentration of delta Di-HA between the PISF and GCF of the controls. The ratios of hyaluronic acid to chondroitin sulfate in the GCF and in the whole saliva of the periodontitis group were significantly lower than that of the controls. There was no difference between the ratios in PISF and those in GCF of the controls. These results indicate that checking hyaluronic acid in GCF and whole saliva using HPLC is a useful means of assessing the condition of periodontal tissues, and that assaying hyaluronic acid in PISF may also be effective for monitoring the condition of tissues around dental implants. PMID- 9872079 TI - Effect of antibiotics on rat leukocyte function. AB - Many antibiotics are used to treat infection in clinical practice. Actions of these drugs involve specific immune enhancement and improved overall phagocytic and bactericidal capacities of polymorphonuclear leukocytes and macrophages at the site of infection. In this study, we examined the effects of antibiotics on chemotaxis and phagocytosis by reacting macrophages with 7 different antibiotics, specifically ampicillin (ABPC), cephalexin (CEX), cefotiam (CTM), amikacin (AMK), clindamycin (CLDM), tetracycline (TC) and bleomycin (BLM). At 1 microgram/ml, there were significant differences in chemotaxis between control and experimental groups treated with agents other than ABPC and CEX (CLDM: p < 0.05, AMK: p < 0.01, CTM, TC and BLM: p < 0.001). At 10 micrograms/ml, there were significant differences in chemotaxis between the control group and all treated groups (ABPC and CLDM: p < 0.01, CEX, CTM, AMK, TC and BLM: p < 0.001). At 100 micrograms/ml, all antibiotics significantly inhibited chemotaxis (p < 0.001). Phagocytosis was evaluated by determining both the ratio and index of phagocytosis. There was a significant difference in the phagocytosis ratio between the control group and the group treated with 10 micrograms/ml of BLM (p < 0.001). At 100 micrograms/ml, all agents but ABPC significantly reduced phagocytosis in a dose-dependent manner. Agents other than ABPC similarly reduced the phagocytosis index. Significant differences were noted between the control group and groups treated with BLM or AMK (BLM: 10 micrograms/ml: p < 0.01, 100 micrograms/ml: p < 0.001, AMK: 100 micrograms/ml: p < 0.01). PMID- 9872080 TI - Differential effects of systemic morphine on responses elicited by tooth pulp stimulation of nociceptive neurons in lateral and medial thalamic nuclei. AB - The effects of intravenous morphine on responses of feline thalamic nociceptive neurons receiving afferent input from the tooth pulp (TP) were investigated. Morphine suppressed responses to TP stimulation in both tooth pulp specific (TPS) and wide dynamic range (WDR) neurons with TP input recorded from the nucleus ventralis posteromedialis (VPM). However, there was scant morphine effect on responses to stimulation of trigeminothalamic tract (TTT) fibers in the trigeminal medial lemniscus. Furthermore, in nociceptive neurons with TP input recorded from nucleus centralis lateralis (CL) and parafascicularis (Pf) of the intralaminar nuclei, intravenous morphine suppressed responses both to stimulation of the mesencephalic reticular formation (MRF) as well as TP stimulation. The suppressive action of morphine on responses elicited by TP stimulation of VPM, CL and Pf neurons was antagonized by intravenous naloxone (1 mg/kg). Results suggest that intravenous morphine suppresses synaptic transmission of nociceptive impulses in the intralaminar nuclei as well as in the lower brain stem but not in the VPM. PMID- 9872081 TI - SEM study on microvascular changes following implantation of bone morphogenetic protein combined with hydroxyapatite into experimental bone defects. AB - BMP-HAP complex was implanted into a bone defect in the femur of Wistar-strain rats, and animals were allowed to heal for one to 8 weeks prior to sacrifice. Similar bone defects without BMP-HAP complex served as controls. Osseous healing and microvascular changes, as revealed by plastic microcorrosion castings, were subsequently examined under a scanning electron microscope. One week after implantation, sproutings and congregate sinusoidal capillary plexuses and primary bone trabeculae (woven bone) were observed around and between the BMP-HAP complexes. Control specimens revealed a fine and immature sinusoidal capillary plexus arising from sproutings and elongations of pre-existing blood vessels, but bone formation was not observed. At two weeks, newly-formed trabeculae were observed around and on the surface of the HAPs, and a network of thick, newly formed vessels was observed in intervening space. At three weeks, networks of newly-formed vessels were observed on the surface of the HAPs, and surrounding newly-formed trabeculae had become thickened. At four weeks, bone defects were filled, and HAP was completely embedded in new bone. At eight weeks, the HAP was fused with new bone, and the boundary between the HAP and new bone was unclear. In comparison with comparable surgically created control bone defects without implanted BMP-HAP complex, BMP apparently stimulates new vascularization. Further, implanted BMP-HAP apparently stimulates undifferentiated mesenchymal cells to differentiate into angioblasts and osteoblasts, or vice versa (dedifferentiation). PMID- 9872082 TI - Immunohistochemical staining of human teeth, and production of monoclonal antibodies against cementum. AB - Influence of various forms of fixation and decalcification on immunohistochemical staining of paraffin-embedded human teeth and surrounding tissues was initially examined using commercially available antibodies against vimentin (V), type I collagen (C) and cytokeratin (K). Secondly, monoclonal antibody (MoAb) was produced against both bovine and human cementum and immunohistochemical screening was subsequently undertaken to test reactivity with different scheme of fixation and decalcification. The combination of neutral buffered paraformaldehyde fixation, Morse solution and unmasking procedure yielded both optimal morphology and immunoreactivity. The application of this method into production of MoAb against human or bovine cementum generated a variety of MoAbs reactive with both human teeth and surrounding tissues. The percentage of hybridoma supernatant reactive with sections of human teeth was 14.0-22.4% and was both higher and much improved compared to results of previous soft tissue studies. Finally, the MoAbs, BC 1 and 2 (isolated following the use of bovine cementum as immunogen) and HC 1 (human cementum immunogen) recognized specific bands of various molecular weight. The three MoAbs showed strong resistance against periodate oxidation and borohydrate reduction and were stable following treatment with proteolytic enzymes. All were immunoreactive with components of cementum. PMID- 9872083 TI - Guided tissue regeneration (GTR) versus cementum-impregnated gelatine membrane (CGM) techniques: a histologic comparison of relative effectiveness in promoting periodontal attachment. AB - Some prior reports have suggested that guided tissue regeneration (GTR) procedures achieve only partial regeneration and induces the ankylosis rather than true attachment. Accordingly, others have developed an alternative procedure employing gelatine membrane compounded with bovine cementum particles (CGM) which has proven effective in stimulating a more physiologic form of attachment. This study was undertaken to perform a direct comparison of histological results when CGM and GTR membrane were used at comparable sites in the same monkey. Three monkeys with no periodontal disease were used. Following flap surgery, recession type defects were created on the buccal side of the maxillary lateral incisors and second premolars, and the cementum was removed from the root surface at an area corresponding to the bone crest. The right and left lateral incisors and second premolars were covered with CGM and GTR membrane, respectively. The GTR membranes were removed after 4 weeks. At 6 wks, the animals were sacrificed, and specimens were prepared for histological examination. More coronally placed true new attachment was observed following application of CGM to the planed root surfaces. Application of the GTR membrane resulted in formation of bone-like cementum and ankylosis, whereas CGM established true periodontal regeneration. PMID- 9872084 TI - Age-related changes in collagen, laminin and tenascin in the infant rat submandibular gland. AB - We investigated biochemically and immunohistochemically the age-related changes in collagen (types I, III and IV), laminin and tenascin of the extracellular matrix in the submandibular gland of infant rats. The hydroxyproline content markedly increased on day 14 after birth. Electrophoretic profile revealed alpha chains of only type I collagen up to day six after birth. Type III collagen, which was diffusely distributed in interlobular connective tissue, was gradually expressed from day seven after birth. Type IV collagen content increased from day seven. Laminin content gradually increased with age. Tenascin immunohistochemically showed weak expression only around the vessel walls up to day seven. In addition, this expression markedly increased around acinar and ductal walls as basement membrane components on day 14 after birth. These findings suggest that rat submandibular gland types I and IV collagen, and laminin are already present during the fetal stage and relate to construction of tissue, and that type III collagen and tenascin, which increased from day seven after birth, relate to maturation. PMID- 9872085 TI - Effects of heat fixation on tissue structure, immunostaining and in situ RT-PCR. AB - The effects of PBS heat fixation on tissue structure, immunostaining, and in situ RT-PCR were investigated using excised mucosal tissues from both humans and various animals. It was found that basal cells were stretched to two to three times their normal length at 60 degrees C, and 4 to 5 times at 90 degrees C, resulting in detachment of the epithelium and basal cells from subjacent connective tissue. Tonofibrils disappeared completely and became an electron dense amorphous mass at 60 degrees C. Both nuclear and cytoplasmic membranes also disappeared as did intracytoplasmic organelles. Electron dense amorphous-material occupied the entire nucleus with fixation at 90 degrees C. Collagen fibrils appeared swollen at 60 degrees C and either disappeared completely or became fragmented undergoing transition to an electron dense amorphous conglomerates at 90 degrees C. Immunostaining of cytokines disappeared at 60 degrees C, and that of vimentin at 90 degrees C. However, cytokeratin and human leukocyte common antigen (CD 45RB) reactivity remained intact at both temperatures. IL-6 mRNA could be localized by in situ RT-PCR even after thermal fixation. Results suggest that a proper combination of immunohistochemistry and in situ RT-PCR will enable investigators to localize proteins in addition to DNA or RNA within oral tissue sections. This capability should prove helpful in applied histopathologic diagnostic techniques. PMID- 9872086 TI - Lectin binding patterns of odontogenic epithelium in the rat during various phases of molar tooth development. AB - Light and electron microscopic investigations of lectin binding patterns in rat tooth germs were undertaken in order to elucidate glycoconjugate localization in cells of the reduced enamel epithelia and their derivatives. It was found that Ulex europaeus agglutinin (UEA-1), peanut agglutinin (PNA), soybean agglutinin (SBA), and wheat germ agglutinin (WGA) exhibited variable reactivity patterns with different epithelia. UEA-1 was reactive with cells of the stratum intermedium and stellate reticulum in the tooth germ but unreactive with ameloblasts, outer enamel epithelial cells, and junctional epithelium at later stages. Reaction patterns of PNA in these cells differed from those of UEA-1. Results indicated that inner and outer cells of the reduced enamel epithelium are heterogeneous with regard to lectin binding patterns. PMID- 9872087 TI - The effect of leukocyte function of streptozotocin-induced diabetes in naturally occurring gingivitis rat. AB - Although diabetes mellitus is known to aggravate periodontal disease, the precise relationship between these two entities is far from being completely understood. Further study of this relationship was therefore undertaken in the form of observation of both naturally occurring gingivitis in rats (ODUS/Odu) and effects produced by induction of experimental diabetes mellitus by injection of streptozotocin (STZ: 65 mg/kg, i.v.). At one and 3 mon after STZ injection, liquid paraffin was injected intraperitoneally. Four days thereafter, pocket probing depths of rats were measured and blood samples as well as peritoneal macrophages were collected from both experimental animals and non diabetic controls. Both chemotaxis and phagocytosis of macrophages were studied. At one and 3 mon after STZ injection, pocket probing depths of diabetic animals were significantly deeper than those of controls (p < 0.001). Pocket probing depths were deeper at 3 mon after STZ injection than after 1 mon in diabetic animals. At three months after STZ injection, there was a high degree of positive correlation between pocket probing depths, blood glucose levels, triglyceride, and hemoglobin A1c levels (p < 0.01). Also, macrophage chemotaxis was more suppressed in diabetic rats than it was in controls. Additionally, both phagocytosis ratios and phagocytosis indices of macrophages in the diabetes group were significantly more suppressed than those in controls in both experimental periods (p < 0.001). Findings suggest that both chemotaxis and phagocytosis are compromised in macrophages from rats rendered diabetic by STZ injection. Thus as host defense mechanisms become weakened, there is a corresponding progression of periodontal disease. PMID- 9872088 TI - Tooth pulp neurons in the caudal medulla oblongata of the cat. AB - The medulla oblongata caudal to the obex was explored for neurons responsive to tooth pulp (TP) stimulation in cats. Four different subclasses of TP neurons were found. The latter included TP specific (TPS) neurons, trigeminal wide dynamic range (trigeminal WDR) neurons with TP input, trigeminal subnucleus reticularis ventralis (trigeminal SRV) neurons with TP input and convergent reticular formation (convergent RF) neurons with TP input. TPS neurons were located in the dorsal marginal rim of the trigeminal subnucleus caudalis, i.e., in the marginal layer or the outer zone of substantia gelatinosa. WDR neurons with TP input were found in the neck region of medullary dorsal horn which corresponds to the lateral part of subnucleus reticularis dorsalis (SRD). Trigeminal SRV neurons with TP input were located in the lateral part of SRV. Convergent RF neurons with TP input were found in the middle third of the caudal bulbar RF consisting of SRD and SRV. Both TPS neurons and WDR neurons with TP input included trigeminothalamic neurons as evidenced by the antidromic activation from the nucleus ventralis posteromedialis of the contralateral thalamus. A significant proportion of both trigeminal SRV and convergent RF neurons with TP input were antidromically activated by stimulation of the nucleus centralis lateralis of the contralateral thalamus. The former two subclasses may subserve the sensory discriminative aspect of toothache, while the latter two subclasses, the emotional-motivational aspect. PMID- 9872089 TI - Effect of particle sizes in India ink on its use in evaluation of apical seal. AB - We investigated the use of India ink as an indicator of root canal sealing ability. Sealing ability is one of the most important factors required of root canal cements. Various dyes have been used for this purpose. Methylene blue, radioisotopes and India ink have all been tried. However, there is no evidence that India ink is suitable as an indicator. We measured the particle size distribution of India ink to determine how this factor affects test results. In addition, we observed the surface texture of an experimentally developed calcium phosphate sealer and a commercially available root canal sealer using SEM. We found that a portion of the India ink particles were smaller than cracks on the two sealer surfaces, indicating, that India ink could pass through these cracks. We concluded that India ink is suitable as an indicator of root canal seal. PMID- 9872090 TI - Happiness is a byproduct of the pursuit of excellence. PMID- 9872091 TI - Antibiotics: indications, contraindications, and non-indications. PMID- 9872092 TI - Workplace violence? It can happen to you, doctor. PMID- 9872093 TI - Current concepts regarding medical necessity. PMID- 9872094 TI - Cashstreams: five powerful income streams to increase your net income. AB - You can dramatically increase your profits by: extending credit and earning interest on the unpaid balance; doing all of the undone treatment in your practice; providing financing for everyone who deserves it; treating bigger cases; and avoid treating deadbeats. These five cash stream strategies will revitalize your practice with more new patients, better case acceptance, and increased cash flow. But bottom line, you must get good at financing. You must have in place an organized, proven, financing system--just like the finance companies do. PMID- 9872095 TI - Cover your health care costs with a medical savings account. PMID- 9872096 TI - Respiratory obstruction and its role in long face syndrome. PMID- 9872097 TI - Exquisite moments of American dental heritage. PMID- 9872098 TI - Increase your awareness of potential violence. PMID- 9872099 TI - Financing college education. AB - You have more options than ever before for financing a college education. You can: Split income with your children Prepay your child's tuition Buy a rental house for your college student Hire your child Establish and educational IRA Deduct interest on some student loans Claim the new tax credits for education. With today's educational cost, you need more possibilities for maximizing your dollars. Keep this article handy. Use it to help you take full advantage of the old, new, and varied possibilities. PMID- 9872100 TI - Late bloomers: saving for college when your kids are teenagers. PMID- 9872101 TI - Collection control in three steps. PMID- 9872102 TI - Reconstruction of the mandibular dentition: the lost arch. PMID- 9872103 TI - Protocol for predictable restoration of anterior teeth with composite resins. AB - The composite resins have reached a high degree of excellence and are now used with predictable results in restoring the anterior dentition. This article discusses the development of these resins and the histologic and optical considerations (polychromatic characteristics, hue translucency and opacity, chroma, value, strength and polishability). In restorative materials, the use of universal or all-purpose restorative resins is considered: i.e. materials that combine the junctional strength necessary for the posterior regions and the high gloss polishability necessary to emulate enamel in the anterior. The restorative sequence of cervical, middle, and incisal thirds describes the materials best suited for each third, along with aids to create the intricate refinement of hue, chroma, and value. The learning objective of this article is to review the principles involved in these restorations and to provide a protocol for their clinical implementation. PMID- 9872104 TI - Significance of the biological width to the restorative dentist. PMID- 9872106 TI - Are you selling your office? PMID- 9872105 TI - Implants in the partially edentulous patient: restorative considerations. AB - Restorative considerations are critical to the long-term success of fixed implant supported prostheses, especially in the posterior quadrants of the partially edentulous patient. The parafunctional habit of bruxism must be identified and addressed. The restoration should dictate implant placement. Control of forces directed upon the prosthesis and implants is critical to long-term success. Anatomic limitations to implant placement and surgical procedures to correct these deficiencies must be considered for their impact on the prosthetic restoration. Nonaxial forces or bending moments should be minimized by the use of an adequate number, position and alignment of implants; by control of the occlusion; and by design of the prosthesis. The patient must understand the risks, limitations, costs and time commitments of implant restorations prior to treatment. PMID- 9872107 TI - Actinomyces naeslundii genospecies 1 and 2 express different binding specificities to N-acetyl-beta-D-galactosamine, whereas Actinomyces odontolyticus expresses a different binding specificity in colonizing the human mouth. AB - A total of 102 strains of Actinomyces were isolated from teeth, buccal mucosa and tongue in eight individuals. The isolates were characterized by multivariate statistical analyses of phenotypic characteristics, serotyping and binding to beta-linked galactosamine (N-acetyl-beta-D-galactosamine) and acidic proline-rich protein structures. Based on these characteristics, isolates were classified into three major groups: (i) Isolates of Actinomyces naeslundii genospecies 2 were the dominant species on teeth and buccal mucosa and bound commonly to N-acetyl-beta-D galactosamine (63 of 63 isolates) and acidic proline-rich proteins (63 of 63 isolates), regardless of tissue origin. They all exhibited a N-acetyl-beta-D galactosamine binding specificity signified by N-acetyl-beta-D-galactosamine inhibitable coaggregation with the streptococcal strains LVG1, GVE1, 24892 and MPB1; (ii) Isolates of A. naeslundii genospecies 1 were prevalent on teeth in certain individuals and bound commonly to N-acetyl-beta-D-galactosamine (20 of 20 isolates), but less commonly to acidic proline-rich proteins (5 of 20 isolates). They all possessed another N-acetyl-beta-D-galactosamine specificity, i.e. N acetyl-beta-D-galactosamine-inhibitable coaggregation with the same streptococcal strains except for strain MPB1; (iii) Isolates of Actinomyces odontolyticus, the dominant species on the tongue (17 of 19 isolates), bound commonly to unknown structures on streptococci (17 of 19 isolates) but rarely to N-acetyl-beta-D galactosamine (2 of 19 isolates) or acidic proline-rich proteins (3 of 19 isolates). In conclusion, A. naeslundii genospecies 1 and 2 exhibit different patterns of N-acetyl-beta-D-galactosamine and acidic proline-rich protein specificities to colonize dental and buccal mucosa surfaces, whereas A. odontolyticus utilizes another specificity to colonize the tongue. PMID- 9872108 TI - Actinomyces serovar WVA963 coaggregation-defective mutant strain PK2407 secretes lactose-sensitive adhesin that binds to coaggregation partner Streptococcus oralis 34. AB - Actinomyces serovar WVA963 strain PK1259 mediates intergeneric coaggregation with several oral streptococci. These lactose-inhibitable coaggregations appear to involve a 95-kDa putative actinomyces adhesin in complex with type 2 fimbriae. A coaggregation-defective strain PK2407 lacking type 2 fimbriae synthesizes the putative adhesin but appears unable to present it properly on its surface. Antiserum was raised against surface sonicates of PK2407 and was absorbed with a different coaggregation-defective mutant PK3092 that synthesizes type 2 fimbriae but no adhesin. This absorbed antiserum specifically blocked lactose-inhibitable coaggregation of wild-type strain PK1259 and Streptococcus oralis 34 and identified a 95-kDa protein in ammonium sulfate precipitates of culture supernatant of the coaggregation-defective mutant PK2407. The 95-kDa secreted protein was bound to the streptococcal partner cells and to lactose-agarose affinity beads and was released by lactose from both the affinity beads and partner, indicating that the secreted and precipitated protein is biochemically active and may mediate coaggregation with streptococci. PMID- 9872109 TI - Epithelial cell invasion by Actinobacillus actinomycetemcomitans strains from restriction fragment-length polymorphism groups associated with juvenile periodontitis or carrier status. AB - The epithelial cell invasiveness of Actinobacillus actinomycetemcomitans strains of different restriction fragment-length polymorphism (RFLP) groups associated with disease conversion and asymptomatic carrier status in localized juvenile periodontitis was examined. Twenty clinical isolates were studied for their ability to invade KB monolayers, using the quantitative gentamicin killing assay. Five isolates were found to be invasive, five were not invasive; and the other 10 did not invade better than an invasion negative control Haemophilus aphrophilus strain ATCC 19415. Using probe-specific DNA fingerprinting. 11 strains were assigned to RFLP group II (disease-associated); 4 to RFLP type XIII (carrier status associated); and the other to groups III, IV, V and VII. Eight isolates, all RFLP group II, were leukotoxin producers as determined by PCR amplification of the lkt promoter region. No correlation was found between invasiveness and RFLP group. Leukotoxin production was more associated with noninvasive than invasive strains. PMID- 9872110 TI - Binding of the capsule-like serotype-specific polysaccharide antigen and the lipopolysaccharide from Actinobacillus actinomycetemcomitans to human complement derived opsonins. AB - We investigated the molecular mechanism of resistance of Actinobacillus actinomycetemcomitans to complement-dependent chemiluminescence response by human polymorphonuclear leukocytes. Whole cells of serotype b-specific polysaccharide antigen-defective mutants ST2 and ST5 were constructed by inserting transposon Tn916 into A. actinomycetemcomitans strain Y4. These strains induced strong chemiluminescence response by human polymorphonuclear leukocytes and markedly bound to human complement-derived opsonins. In contrast, strain Y4 induced weak chemiluminescence response and weakly bound to complement-derived opsonins. The biosensor analysis revealed that lipopolysaccharide from strain Y4 strongly bound to human C3b, but serotype b-specific polysaccharide antigen did not. The serotype b-specific polysaccharide antigen molecule might sterically hinder the interaction between complement-derived opsonins and lipopolysaccharide to reduce complement-dependent chemiluminescence response by human polymorphonuclear leukocytes. PMID- 9872111 TI - Antigen-specific T-cell receptor V beta expression in Porphyromonas gingivalis specific T-cell lines. AB - FACS analysis was used to determine the expression of 15 T-cell receptor V beta families on CD4 and CD8 cells in Porphyromonas gingivalis specific T-cell lines established from eight P. gingivalis-positive adult periodontitis and seven P. gingivalis-positive healthy or gingivitis subjects. All 15 T-cell receptor V beta families were expressed by the T-cell lines, although a significantly higher proportion of the CD4 cells expressed the 5.2-3 V beta region compared with the other 14 families, including the 5.3 region, suggesting that it is the 5.2 family which is overexpressed. This was also true for the CD8 cells, with the exception of the 3.1 region in adult periodontitis T-cell lines and the 3.1, 13.1/13.3 and 21.3 regions in healthy or gingivitis lines. Between the two clinical groups, a significantly lower percentage of 13.1/13.3-positive CD8 cells was noted in the adult periodontitis lines compared with the healthy or gingivitis lines. There was a significant reduction in DNA synthesis by the lines in the presence of P. gingivalis outer membrane antigens and fixed irradiated lymphoblastoid cell lines compared with cultures containing untreated irradiated lymphoblastoid cell lines and in cultures containing anti-class II major histocompatibility complex antibody in comparison with all other cultures. The results of this study have shown that P. gingivalis preferentially induces the T-cell receptor V beta 5.2 family on CD4 and CD8 cells in P. gingivalis-specific T-cell lines and that activation of T cells by P. gingivalis outer membrane antigens may be by antigen specific rather than superantigen activity. PMID- 9872112 TI - A hemagglutinating variant of Prevotella melaninogenica isolated from the oral cavity. AB - Strains resembling Prevotella melaninogenica were isolated from healthy subjects and patients with periodontal disease and were identified using: a 5-test phenotypic screen; commercial identification kits; and a 16S rRNA-based polymerase chain reaction (PCR) method. Eleven clinical isolates closely resembling P. melaninogenica, and all from patients with periodontitis, were able to agglutinate erythrocytes. In the electron microscope, hemagglutinating isolates showed fimbria-like structures, that were not seen on non hemagglutinating isolates. Some strains were further classified with PCR restriction fragment-length polymorphism (RFLP) of 16S rRNA genes. Amplified 16S rDNA was digested using five different endonucleases, separated with agarose gel electrophoresis, stained and photographed. Photographs were then scanned, digitized and a distance matrix calculated using Dice coefficient, where the presence or absence of a band was used as a character. The distance matrix was plotted as a phenogram. At 70% similarity six clusters were seen. Type strains of separate Prevotella species did not fall into any cluster. Hemagglutinating isolates fell into three clusters: four clustered with the type strains of P. melaninogenica and Prevotella veroralis; four with other P. melaninogenica isolates and two hemagglutinating isolates clustered together Prevotella loescheii. The PCR-RFLP results showed that the hemagglutinating strains did not form a homogenous group inside the Prevotella genus. PMID- 9872113 TI - Pathogenicity of facultative and obligate anaerobic bacteria in monoculture and combined with either Prevotella intermedia or Prevotella nigrescens. AB - The pathogenicity of obligate and facultative anaerobic bacteria commonly found in endodontic infections was tested using a mouse model. The capacity of inducing abscesses was evaluated seven days after subcutaneous injection of the bacteria in pure culture and in combinations with either Prevotella intermedia or Prevotella nigrescens. Nine of the fifteen bacterial strains tested were pathogenic in pure culture. No statistically significant differences were detected between these strains in pure culture and in mixtures with either P. intermedia or P. nigrescens. Synergism between the bacterial strains was only apparent when associating Porphyromonas endodontalis with P. intermedia or P. nigrescens. Histopathological examination of tissue sections from induced abscesses revealed an acute inflammatory reaction, dominated by polymorphonuclear leukocytes. Sections from the control group using sterile medium showed no evidence of inflammatory reaction. PMID- 9872114 TI - Virulence of a polymicrobic complex, Treponema denticola and Porphyromonas gingivalis, in a murine model. AB - The effect of a polymicrobic infection employing Treponema denticola and Porphyromonas gingivalis in the murine lesion model was used to determine the synergistic virulence of these two periodontopathic bacteria. At high doses of P. gingivalis W50, addition of T. denticola in the infection mixture had no effect on the formation and size of the spreading lesion caused by this microorganism. However, at low P. gingivalis challenge doses, T. denticola significantly enhanced the virulence of P. gingivalis compared with monoinfection of this microorganism. A potential role of the trypsin-like protease enzyme activity of P. gingivalis in this synergistic virulence was tested using P. gingivalis mutants deficient (i.e., BEI) or devoid (i.e., NG4B19) of this protease activity. These findings demonstrated that T. denticola-P. gingivalis complexes exhibit enhanced virulence in this model and that even using a polymicrobic challenge infection, the trypsin-like protease activity was important to P. gingivalis virulence expression. PMID- 9872115 TI - Combined inhibitory effect of bovine immune whey and peroxidase-generated hypothiocyanite against glucose uptake by Streptococcus mutans. AB - Immune whey product was obtained from Streptococcus mutans- and Streptococcus sobrinus-immunized cows. Hypothiocyanite (HOSCN/OSCN-) was generated in VMG buffer, pH 5.5 or 6.5, by bovine milk lactoperoxidase, KSCN and hydrogen peroxide. The glucose incorporation by late-log cells of S. mutans 10449, serotype c, was followed by measuring the uptake of 14C-glucose at 37 degrees C. At pH 5.5 and 6.5 both immune whey product and HOSCN/OSCN- dose-dependently inhibited glucose uptake. The inhibition by their combination was additive if bacterial cells were treated with HOSCN/OSCN- before exposed to immune whey product. In contrast to immune whey product, the control product from sham immunized cows increased the glucose uptake even when added simultaneously with HOSCN/OSCN-. However, when bacterial cells were pretreated with HOSCN/OSCN- an enhanced inhibitory effect was observed also with control product. The results indicate that colostral proteins from S. mutants- or S. sobrinus-immunized cows inhibit glucose uptake and that the effect is enhanced by pretreatment with lactoperoxidase-generated HOSCN/OSCN-. PMID- 9872117 TI - Purification and properties of a bacteriocin of Staphylococcus epidermidis isolated from dental plaque. AB - An extracellular bacteriocin of Staphylococcus epidermidis isolated from dental plaque was purified and characterized. Its molecular mass was 3500 Da and pI was 10.5. This bacteriocin inhibited the growth of Staphylococcus aureus, Streptococcus salivarius, and Streptococcus mitis, but Streptococcus mutans, Streptococcus sanguis and other oral indigenous bacterial species examined were not inhibited. The mode of inhibition was found to be bacteriostatic. PMID- 9872116 TI - Detection of dextranase-producing gram-negative oral bacteria. AB - Thirty-one strains of 23 gram-negative oral bacterial species were examined for dextran-degrading activity on agar plates containing blue dextran. One strain each of Capnocytophaga ochracea, Capnocytophaga sputigena, Prevotella loescheii, Prevotella melaninogenica and Prevotella oralis had detectable dextranase activity. The culture supernatants of P. melaninogenica and P. oralis cells contained dextranases of multiple sizes, but those of the other three species had a single size of enzyme. A 56-kDa dextranase was purified from the culture supernatant of P. oralis and the antiserum against the enzyme was prepared with a rabbit. The Ouchterlony test showed that the antibody reacted with the supernatants of both P. melaninogenica and P. oralis but not with the others. Dot blot hybridization using the dextranase gene of Streptococcus mutans as a probe revealed that there was no significantly homologous sequence in the chromosomal DNA of the five species. PMID- 9872118 TI - Dental school debt and generational perception. PMID- 9872121 TI - A kit for financial freedom. PMID- 9872119 TI - New dentists confront serious financial obstacles. PMID- 9872120 TI - Analysis of dental incomes reveals disturbing trends. PMID- 9872122 TI - Management service organizations: consolidation sparks debate for dentists. PMID- 9872123 TI - Is there anything in it for you? ADA experts explain management service organizations. PMID- 9872124 TI - MSO bankruptcy tangles dentists in legal red tape. PMID- 9872125 TI - Factors influencing the immediate plans of the 1997 graduating class. PMID- 9872126 TI - St. Paul's Dental Clinic fills valuable role. PMID- 9872127 TI - An embarrassment of choices. PMID- 9872128 TI - The relation between otoacoustic emissions and the broadening of the auditory filter for higher levels. AB - The active behaviour of outer hair cells (OHCs) is often used to explain two phenomena, namely otoacoustic emissions (OAEs) and the level dependence of auditory filters. Correlations between these two phenomena may contribute to the evidence of these hypotheses. In this study auditory filters were calculated from probe thresholds in notched-noise maskers over a range of at least 25 dB. Further. transient evoked otoacoustic emissions (TEOAEs) were measured at several stimulation levels. Ten normal-hearing and nine hearing-impaired subjects were tested. A linear increase of the width of the auditory filter with 2.2 dB/Hz was found up to a specific saturation level. The group of selected hearing-impaired subjects with mild hearing loss showed no wider than normal auditory filters. As expected, the increase of the width of the auditory filter correlated positively with the level of TEOAEs for click intensities of about 80 dB peak SPL. However, for subjects with TEOAEs wider auditory filters at a masker level of 65 dB/Hz were found for subjects with larger TEOAEs. This result cannot be explained by a model by which the cochlea shows an active behaviour for lower stimulation levels, influencing both the TEOAE levels and the filter skirts, and a passive behaviour for higher stimulation levels. PMID- 9872129 TI - Endonuclease cleavage of DNA in the aged cochlea of Mongolian gerbil. AB - Apoptosis is the most common form of physiological cell death, plays an important role in a variety of physiological and pathological situations, and is characterized by an endonuclease activation. Aged gerbils (24-month-old) showed a significant elevation of the distortion product otoacoustic emission (DPOAE) threshold and a significant decrease of the DPOAE level as compared to young gerbils (3- to 6-month-old), suggesting a disturbance of the outer hair cell in the aged cochlea. Furthermore, the endocochlear potential in the aged (15-75 mV) was apparently lower than that in the young (80-90 mV), indicating strial damage with aging. A terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method demonstrated the presence of the DNA fragmentation predominantly in the outer hair cells and spiral ganglion cells. These changes were sporadically observed in the inner hair cells and supporting cells. TUNEL positive cells were markedly observed in some aged gerbils showing deterioration in cochlear function, which may suggest the individual variability of presbycusis. On the other hand, the strial cells showed a marked atrophy, but no DNA fragmentation. These findings suggest that the aged change in the outer hair cells and spiral ganglion cells involves an apoptotic death pathway. However, strial atrophy with aging seems to be unrelated to apoptosis. PMID- 9872130 TI - Tonotopic organization of the human auditory cortex as detected by BOLD-FMRI. AB - Functional magnetic resonance imaging is a noninvasive and nonradioactive method for the detection of focal brain activity. In the present study the auditory cortex was investigated in nine normal subjects who were binaurally stimulated using pulsed sine tones of 500 Hz and 4000 Hz. The BOLD (blood oxygenation level dependent) signal change coincided with the stimulation paradigm and was detected in the plane of the superior temporal gyrus. The comparison of the spatial distribution of activated areas revealed a different behavior for the two frequencies. The present findings underline the existence of a frequency specific organization in the medio-lateral, fronto-occipital and cranio-caudal extension in both hemispheres of the auditory cortex in human. The activated areas for the high tone were found more frontally and medially orientated than the low tone stimulated areas. Furthermore, a slight cranio-caudal shift was observed for the higher frequency, more pronounced in the right than in the left temporal lobe. Finally, for most of the subjects investigated the BOLD activation area of the 500 Hz sine tone was larger than that of the 4000 Hz stimulation. Both frequencies showed a lateralization of signal response to the left temporal lobe. PMID- 9872131 TI - The effect of mannitol upon cochlear dysfunction induced by transient local anoxia. AB - Transient local anoxia of the cochlea was induced by pressing the labyrinthine artery, and compound action potential (CAP) or endocochlear potential (EP) was measured before and after transient local anoxia ranging from 5 to 60 min using 106 albino guinea pigs. The complete interruption of the cochlear blood flow by this procedure and its full restoration after releasing the pressure on the artery was confirmed by a laser-Doppler flowmeter. The anoxia of less than 10 min induced no post-anoxic cochlear dysfunction, whereas the anoxia of a longer duration induced an irreversible dysfunction of the cochlea. It was evident that the post-anoxic recovery of the CAP threshold was worse as the anoxia period was prolonged, and CAP was almost completely abolished after 60-min anoxia. In animals which were administered mannitol intravenously just after the restoration of the cochlear blood circulation, the recovery of the CAP threshold was significantly better than that in the control animals, when the animals were subjected to local anoxia of 15- to 30-min duration. No beneficial effect, however, was observed in the 60-min anoxia group. In conclusion, local anoxia of 10 min or longer caused cochlear dysfunction, which was partially but significantly alleviated by mannitol. PMID- 9872132 TI - OCP2 exists as a dimer in the organ of Corti. AB - OCP2 is one of the most abundant proteins in the organ of Corti (OC), comprising approximately 5% of the total protein in the supporting cell population. Although the very close homolog, Skp1p, has been implicated in regulating cell-cycle progression, the function of OCP2 in the terminally differentiated cochlea is presently unknown. We have purified recombinant OCP2 from Escherichia coli and examined the protein by analytical ultracentrifugation. Interestingly, sedimentation equilibrium data collected at 20 degrees C unequivocally indicate that, at the concentrations present in the OC, free OCP2 would exist as a dimeric species. The apparent sedimentation coefficient is independent of concentration at loading concentrations between 10 and 100 microM, indicating the absence of a significant monomer-dimer equilibrium in this concentration range. The functional significance of this finding is discussed. PMID- 9872133 TI - Morphological polarizations of sensory hair cells in the three otolithic organs of a teleost fish: fluorescent imaging of ciliary bundles. AB - It has been suggested that the morphological polarity of sensory hair cells in the otolithic organs plays important roles in directional hearing in fish. In this study, we examined the hair cell polarization patterns in the saccule, utricle, and lagena of a teleost fish, the sleeper goby (Dormitator latifrons). In contrast to using traditional scanning electron microscopy, we employed a simple and rapid method that enabled us to map the hair cell polarization patterns using immunocytochemical and confocal imaging techniques. The hair cells in the caudal part of the saccular epithelium are oriented dorsally and ventrally, with some variations in the caudal end. The hair cells in the rostral part have diverse morphological polarizations. The utricular hair cells fall into internal and external groups which have opposing polarizations. The lagenar hair cells are found in anterior and posterior groups with approximately opposite polarizations. The saccular and lagenar epithelia are oriented perpendicular to the horizontal plane of the fish, while the utricular epithelium lies on the horizontal plane. Thus, the sleeper goby's ear is morphologically capable of being a three-dimensional sound detector. PMID- 9872134 TI - Gender distinctions and lateral asymmetry in the low-level auditory brainstem response of the human neonate. AB - Threshold measures of auditory brainstem response (ABR) were generated in 72 full term newborn infants in response to clicks and tone burst stimuli between 500 and 8000 Hz as detailed in a previous study. These results were further analyzed for differences in response related to ear (lateral asymmetry) and subject gender. Thresholds obtained in male infants were significantly lower than those of females (P = 0.0485). The greatest differences in threshold between male and female infants occurs in the right ear (7.45 dB) as opposed to the left ear (1.56 dB). Both male and female infants have significantly larger wave V amplitude elicited from the right ear than the left (P = 0.0002) using low-level stimuli. Also, as has been noted in adults, female infants have larger amplitude ABRs than males (P = 0.0018), but amplitude differences across gender are significant only in the right ear (ear by gender interaction P = 0.0278). Results of this study indicate that gender differences and lateral asymmetry in auditory function are not a result of gender bias for or unbalanced auditory trauma, but a biologically significant phenomenon that is present at birth. The argument is made that superior right ear performance may be part of cerebral laterality in auditory function. PMID- 9872135 TI - Cochlear outer hair cell electromotility can provide force for both low and high intensity distortion product otoacoustic emissions. AB - It is generally believed that the force for the otoacoustic emission (OAE) generation is provided by a mechanism of electromotility, observed in isolated cochlear outer hair cells (OHCs). OHC electromotility is resistant to several ototoxic reagents, it does not depend on ATP hydrolysis, but it can be blocked by specific sulfhydryl reagents: p-chloromercuriphenylsulfonic acid (pCMPS) and p hydroxymercuriphenylsulfonic acid (pHMPS). We have used these reagents to test whether they also affect OAE. Application of pCMPS and pHMPS on the round window membrane of anesthetized guinea pigs produced a dose-dependent inhibition of the cubic (2F1-F2) distortion product OAE (DPOAE). The inhibition developed progressively from high to low frequencies, reflecting the diffusion of the drugs through the cochlear compartment. The effect of pCMPS and pHMPS was different from the effects of furosemide and lethal anoxia, which impair cochlear function but do not block OHC electromotility. pHMPS suppressed DPOAE completely at all sound intensities tested (45-80 dB SPL), whereas furosemide or lethal anoxia caused DPOAE to disappear at low-level stimulation (45-60 dB SPL) only. Our results suggest that the OHC electromotility might provide the force for DPOAE generation not only at low, but also at high stimulus intensities. PMID- 9872136 TI - A frequency-dependent saturation evident in rate-intensity functions of the chinchilla auditory nerve. AB - The shape of rate-intensity functions recorded from individual neurons of the auditory nerve using stimulus frequencies at and below the characteristic frequency have been both well-characterized and modeled by other researchers. However, previous studies of rate-intensity functions using stimulus frequencies above the characteristic frequency have primarily focused on the slopes of the rising phases of the functions. Hence, they did not determine whether rate intensity functions recorded using stimulus frequencies above the characteristic frequency saturate, and, if so, at what firing rates the saturation occurs. In this study, rate-intensity functions have been obtained from neurons of the eighth nerve of the chinchilla in response to gated, sinusoidal stimuli in order to investigate saturation firing rates for frequencies above the characteristic frequency. For each neuron, rate-intensity functions were obtained for stimulus intensities up to 90 dB SPL at the characteristic frequency and at as many frequencies above the characteristic frequency as time would allow. These data clearly reveal that, for frequencies above the characteristic frequency, saturation occurs at a rate that decreases monotonically as the frequency of stimulation is increased. In addition, an empirical equation is given which summarizes the dependence of saturation on stimulus frequency for the data of this study. PMID- 9872137 TI - Evidence for loss and recovery of chick brainstem auditory neurons during gentamicin-induced cochlear damage and regeneration. AB - It is well documented that damage to the chick cochlea caused by acoustic overstimulation or ototoxic drugs is reversible. Second-order auditory neurons in nucleus magnocellularis (NM) are sensitive to changes in input from the cochlea. However, few experiments studying changes in NM during cochlear hair cell loss and regeneration have been reported. Chicks were given a single systemic dose of gentamicin, which results in maximal hair cell loss in the base of the cochlea after 5 days. Many new hair cells are present by 9 days. These new hair cells are mature but not completely recovered in organization by 70 days. We counted neurons in Nissl-stained sections of the brainstem within specific tonotopic regions of NM, comparing absolute cell number between gentamicin- and saline treated animals at both short and long survival times. Our data suggest that neuronal number in rostral NM parallels hair cell number in the base of the cochlea. That is, after a single dose of gentamicin, we see a loss of both cochlear hair cells and NM neurons early, followed by a recovery of both cochlear hair cells and NM neurons later. These results suggest that neurons, like cochlear hair cells, can recover following gentamicin-induced damage. PMID- 9872138 TI - Fine structure of the basilar papilla of the emu: implications for the evolution of avian hair-cell types. AB - The morphology of the basilar papilla of the emu was investigated quantitatively with light and scanning electron microscopical techniques. The emu is a member of the Paleognathae, a group of flightless birds that represent the most primitive living avian species. The comparison of the emu papilla with that of other, more advanced birds provides insights into the evolution of the avian papilla. The morphology of the emu papilla is that of an unspecialised bird, but shows the full range of features previously shown to be typical for the avian basilar papilla. For example, the orientation of the hair cells' sensitive axes varied in characteristic fashion both along and across the papilla. Many of the quantitative details correlate well with the representation of predominantly low frequencies along the papilla. The most distinctive features were an unusually high density of hair cells and an unusual tallness of the hair-cell bodies. This suggests that the evolution of morphologically very short hair cells, which are a hallmark of avian papillae, is a recent development in evolution. The small degree of differentiation in hair-cell size contrasts with the observation that a significant number of hair cells in the emu lack afferent innervation. It is therefore suggested that the development of functionally different hair-cell types in birds preceded the differentiation into morphologically tall and short hair cells. PMID- 9872139 TI - Effects of columella removal on inner ear morphology in the chick. AB - We are currently removing the single middle ear bone (columella) in the domestic chick to introduce chemical agents directly into the inner ear. Since we are interested in the effect of these agents on neural structures within the avian basilar papilla (BP), we are concerned about any subtle changes that might result from the surgical procedure of columella removal alone. The purpose of this study was to use light and transmission electron microscopy to analyze morphological changes in the inner ear after columella removal. Fifteen-day-old chicks underwent a unilateral, bilateral or a sham removal of the columella. After columella removal, the oval window was either plugged with Gelfoam or Kimwipe (standard accepted procedure to prevent possible perilymph leak) or left uncovered. After a 5-day survival period, morphological changes were observed in the tegmentum vasculosum (TV) of all ears receiving a columella removal as compared to unoperated ears. Further, ears with Gelfoam plugging the oval window also had damage to the hair cells and support cells of the basilar papilla. In contrast, there were no observable differences in either auditory afferent or efferent nerve terminals on hair cells in the BP from any ears that had the columella removed compared to those from unoperated ears. These results suggest that columella removal alone may produce morphological changes to the TV within 5 days of surgery but not to structures within the BP. On the other hand, columella removal with a Gelfoam plug results in damage not only to the TV but also to cells within the basilar papilla during this same survival time. Despite damage to other structures within the inner ear, cochlear efferent and afferent terminals on surviving hair cells were unaffected by columella removal with or without plugging. PMID- 9872140 TI - Otoacoustic emission in myasthenia gravis patients and the role of efferent activation. AB - We performed transient evoked otoacoustic emission (TEOAE) measurements on 29 ears of myasthenia gravis (MG) patients. The purpose of the study was to support the role of acetylcholine (ACh) in the efferent innervation of cochlear outer hair cells (OHCs). Another aim was to establish additional diagnostic tools for the early determination of MG. Initially, threshold audiometry and impedance measurements showed normal values on the ears examined. The main finding was that TEOAE values were significantly lower in MG patients than in healthy controls. Mestinon, a reversible cholinesterase inhibitor, resulted in a significant increase in mean values of TEOAEs, although these values were still lower than normal. The results suggest that in MG, acetylcholine receptor (AChR) autoantibodies inhibit the function of OHC AChRs. Thus, the TEOAE generated by the active movements of OHCs is decreased in MG. Mestinon prevents the degradation of ACh, and thus stimulates efferent function and increases TEOAE values. The results obtained in this study support the role of ACh in the efferent function of OHC, as well as the impaired function of hair cell AChRs in MG patients. Consequently, measuring TEOAEs may be useful in the early diagnosis of some forms of MG. These results reinforce the importance of collaboration between neurologists and otolaryngologists in the management of diseases with pathological neurotransmission. PMID- 9872141 TI - Lateral suppression of rhythmic evoked responses in the dolphin's auditory system. AB - In the auditory system of bottlenose dolphins (Tursiops truncatus), a brain evoked response to rhythmic sound amplitude modulations (the envelope-following response) was markedly suppressed by addition of another sound with a frequency 5 20 kHz higher and an intensity down to 40 dB lower than that of the amplitude modulated signal. This effect was called paradoxical lateral suppression. This phenomenon was primarily observed when the amplitude-modulated stimulus had a carrier frequency above 30 kHz and modulation rates above 500 Hz. Only the sustained rhythmic response was suppressed, while the transient on-response was not. This indicates that the suppression influenced the ability of evoked potentials to follow rapid amplitude modulations. This prevents weak sounds from being masked by stronger ones. PMID- 9872142 TI - Auditory frequency discrimination in the white rat. AB - Frequency discrimination was investigated in the albino rat using a modified go/no-go positive reinforcement procedure in which subjects reported frequency increments in an ongoing series of pure tone bursts. Weber ratios (frequency difference limen/frequency) were measured from 5 to 32 kHz at 50 dB sound pressure level. A signal detection analysis of the procedure enabled a direct comparison to be made with the rat's performance in a discrete trial go/no-go task. A mean Weber ratio of 3.06+/-0.44% was measured in the frequency range 5-32 kHz. This indicates that the rat has better frequency discrimination acuity than has previously been thought. The result is discussed in the context of factors affecting performance. Among the factors that were explored we found that long training times and the specific training paradigm played important roles. In comparison to discrete trial go/no-go paradigms, rats performed much better when detecting signals from a repeating background. Frequency discrimination performance decreased linearly for tones less than 50 ms in duration. For longer tone duration performance was unaffected. The means and variability of reaction times for threshold changes of frequency were greater in comparison with supra threshold frequency changes. PMID- 9872143 TI - Ultrastructural localisation of spectrin in sensory and supporting cells of guinea-pig organ of Corti. AB - Spectrin is a cytoskeletal protein found in the cortex of many cell types. It is known to occur in cochlear outer hair cells (OHCs) with previous immunoelectron microscopical studies showing that it is located in the cuticular plate and the cortical lattice. The latter is a network of filaments associated with the lateral plasma membrane that is thought to play a role in OHC motility. Spectrin has also been found in inner hair cells (IHCs) and supporting cells using immunofluorescent techniques, but its ultrastructural distribution in these cells has not yet been described. This has, therefore, been investigated using a monoclonal antibody to alpha-spectrin in conjunction with pre- and post-embedding immunogold labelling for transmission electron microscopy. Labelling was found in a meshwork of filaments beneath the plasma membranes of both IHCs and supporting cells and, in pillar cells, close to microtubule/microfilament arrays. It was also found in association with the stereocilia of OHCs and IHCs and, as expected, in the cortical lattice and cuticular plate of OHCs. Thus, spectrin is a general component of cytoskeletal structures involved in maintaining the specialised cell shapes in the organ of Corti and may contribute to the mechanical properties of all the cell types examined. PMID- 9872144 TI - Processing of sinusoidally frequency modulated signals in the nuclei of the lateral lemniscus of the big brown bat, Eptesicus fuscus. AB - Neurons in the nuclei of the lateral lemniscus (NLL) of the big brown bat, Eptesicus fuscus, show several distinctive patterns of response to unmodulated tones. Previous work suggests that sustained responders are specialized to transmit information about sound level and duration while onset responders transmit precise timing information. The biosonar signals of E. fuscus consist of multiple, downward frequency modulated sweeps that change in slope and repetition rate as the bat approaches a target. An obvious hypothesis would be that NLL neurons with sustained responses should discharge during the time when the frequency of a signal is within their response area, but that onset responders should discharge each time the frequency enters the excitatory portion of their response area. In this study we examined the responses of NLL neurons to sinusoidally frequency modulated (SFM) signals presented monaurally to awake, restrained bats. Extracellular recordings were obtained from single neurons in the multipolar and columnar divisions of the ventral nucleus (VNLLm and VNLLc), the intermediate nucleus (INLL) and the dorsal nucleus of the lateral lemniscus (DNLL). All NLL neurons responded synchronously to SFM signals under some conditions. The temporal precision of synchronization was quantified using a coefficient of synchronization (CS), where a value of I equals perfect synchrony. Maximum CS values ranged from 0.70 to >0.99, were generally highest at low modulation rates ( <200 Hz), and showed lowpass characteristics for modulation rate. The maximal modulation rates that elicited synchronous discharge ranged from 50 to 500 Hz. The highest maximal rates were found in the VNLLm and VNLLc, the lowest in DNLL. The ability of NLL neurons to synchronize their discharge to the pattern of an SFM signal is intermediate between that of neurons in the cochlear nucleus and in the inferior colliculus. For the majority of neurons in VNLLm, INLL and DNLL, the precision of synchronization was approximately equal for the downward and upward components of the SFM signal; in contrast, 69% of VNLLc neurons responded selectively to the downward component of the SFM signal. All VNLLc neurons and a subset of those in VNLLm, INLL, and DNLL responded synchronously to SFM signals only if the frequency excursions included a border of the excitatory frequency bandwidth, suggesting that the synchronous discharge was due primarily to the repeated passage of the stimulus frequency into and out of the excitatory portion of the response area. In the case of VNLLc neurons, only the high frequency border was effective; Other neurons, especially those in DNLL, responded synchronously to SFM signals with frequency excursions that were confined entirely within the excitatory response area. PMID- 9872145 TI - Processing of sinusoidally amplitude modulated signals in the nuclei of the lateral lemniscus of the big brown bat, Eptesicus fuscus. AB - Changes in amplitude are a characteristic feature of most natural sounds, including the biosonar signals used by bats for echolocation. Previous evidence suggests that the nuclei of the lateral lemniscus play an important role in processing timing information that is essential for target range determination in echolocation. Neurons that respond to unmodulated tones with a sustained discharge are found in the dorsal nucleus (DNLL), intermediate nucleus (INLL) and multipolar cell division of the ventral nucleus (VNLLm). These neurons provide a graded response over a broad dynamic range of intensities, and would be expected to provide information about the amplitude envelope of a modulated signal. Neurons that respond only at the onset of a tone make up a small proportion of cells in DNLL, INLL and VNLLm, but are the only type found in the columnar division of the ventral nucleus (VNLLc). Onset neurons in VNLLc maintain a constant latency across a wide range of stimulus frequencies and intensities, thus providing a precise marker for when a sound begins. To determine how these different functional classes of cells respond to amplitude changes, we presented sinusoidally amplitude modulated (SAM) signals monaurally to awake, restrained bats and recorded the responses of single neurons extracellularly. There were clear differences in the ability of neurons in the different cell groups to respond to SAM. In the VNLLm, INLL and DNLL, 90% of neurons responded to SAM with a synchronous discharge. Neurons in the VNLLc responded poorly or not at all to SAM signals. This finding was unexpected given the precise onset responses of VNLLc neurons to unmodulated tones and their ability to respond synchronously to sinusoidally frequency modulated (SFM) signals. Among neurons that responded synchronously to SAM, synchronization as a function of modulation rate described either a bandpass or a lowpass function, with the majority of bandpass functions in neurons that responded to unmodulated tones with a sustained discharge. The maximal modulation rates that elicited synchronous responses were similar for the different cell groups, ranging from 320 Hz in VNLLm to 230 Hz in DNLL. The range of best modulation rates was greater for SAM than for SFM; this was also true of the range of maximal modulation rates at which synchronous discharge occurred. There was little correlation between a neuron's best modulation rate or maximal modulation rate for SAM signals and those for SFM signals, suggesting that responsiveness to amplitude and frequency modulations depends on different neural processing mechanisms. PMID- 9872146 TI - Evidence for three additional P2X2 purinoceptor isoforms produced by alternative splicing in the adult rat vestibular end-organs. AB - P2X2 receptors are ligand-gated ion channels that are activated by extracellular ATP. To characterize the expression of P2X2 purinoceptor in the adult rat vestibular periphery, reverse transcription-polymerase chain reaction (RT-PCR) was used. No transcript for P2X2 receptor was found in the vestibular primary afferent neurons (Scarpa's ganglia); however, partial cDNAs encoding four splice variants of the P2X2 receptor were isolated from vestibular end-organs. In all four cDNAs, the deletions were of different lengths but started at the same position on the P2X2 gene (Val-370 codon) located toward the intracellular carboxyl terminus. One of these receptor isoforms was identical in sequence to the recently published P2X2(b) receptor (Simon et al., 1997, Mol. Pharmacol. 52, 237-248) (also known as P2X2-2, in the nomenclature of Brandle et al., 1997, FEBS Lett. 404, 294-298). The remaining three novel splice variants of the P2X2 receptor were designated P2X2(e), P2X2(f) and P2X2(g) (GenBank accession numbers AF028603, AF028604 and AF028605, respectively). The functional significance of these three splice variants remains to be determined. Pituitary and cerebellum were used as survey tissues and only the P2X2(b) receptor cDNA was found. PMID- 9872147 TI - Assessing the role of anti-hsp70 in cochlear impairment. PMID- 9872148 TI - Acupuncture. The West gets the point. PMID- 9872149 TI - Clinical pharmacology. Blue-chip technology. PMID- 9872150 TI - Contraception. Slow train gathers speed. PMID- 9872151 TI - Dementia. It's getting better all the time. PMID- 9872152 TI - Diabetes. Exploding type II. PMID- 9872153 TI - Elderly care. Shifting thinking about memory impairment. PMID- 9872154 TI - Emergency medicine. Accidents will happen? PMID- 9872155 TI - Endocrinology. Can we escape our genetic destiny? PMID- 9872156 TI - Environmental health. More controversy, little clarification. PMID- 9872157 TI - Foodborne illnesses. Strategies for surveillance and prevention. PMID- 9872158 TI - Fraud in medicine. Coping with fraud. PMID- 9872159 TI - Forensic psychiatry. A chill wind blows. PMID- 9872160 TI - Haematology. Important steps forward. PMID- 9872161 TI - Health and human rights. An international minefield. PMID- 9872162 TI - Hepatitis. Advances in antiviral therapy for hepatitis. PMID- 9872163 TI - HIV and AIDS. Gap between biology and reality in AIDS. PMID- 9872164 TI - Imaging. Improving healthcare delivery. PMID- 9872165 TI - Infectious disease. Return of the plagues. PMID- 9872166 TI - Medical education. Training the trainers. PMID- 9872167 TI - Nephrology. Reducing mortality and increasing awareness. PMID- 9872168 TI - Neurovirology. Microbes and the brain. PMID- 9872169 TI - Obstetrics. Preterm birth and pre-eclampsia--bad news and good news. PMID- 9872170 TI - Paediatrics. Science and headlines--controversies continue. PMID- 9872172 TI - Rheumatology. Biological agents--is the promise realised? PMID- 9872171 TI - Perinatal testing. Testing times for pregnant women. PMID- 9872174 TI - Blinded reviews. PMID- 9872175 TI - The AtBCs of lung cancer screening. PMID- 9872173 TI - Xenotransplantation. The viral issue. PMID- 9872176 TI - Reversal of the anticoagulant effects of warfarin by vitamin K1. PMID- 9872177 TI - Rapid vasomotor effects of estrogen: men are part of the club. PMID- 9872178 TI - Pneumonia in Japan: another piece to a worldwide puzzle. PMID- 9872179 TI - Monitor wizards can be dangerous. PMID- 9872180 TI - Insulin without injections??? What have you been puffing? PMID- 9872182 TI - Lung cancer patient follow-up: motivation of thoracic surgeons. AB - STUDY OBJECTIVES: To analyze variation in beliefs that potentially motivate thoracic surgeons in the design of posttreatment surveillance strategies for lung cancer patients and to examine the relationship between motivation and follow-up intensity. DESIGN: International survey. SETTING: Ambulatory care. PARTICIPANTS: All 3,700 members of the Society of Thoracic Surgeons were surveyed to measure their follow-up practices during the 5-year period after treatment, physician beliefs, and variation in these beliefs. The relationship between beliefs, as potentially motivating factors, and follow-up intensity was also analyzed. MEASUREMENTS AND RESULTS: Age, General Thoracic Surgery Club membership, percentage of practice that was noncardiac, South Central United States practice location, and overseas practice location were most frequently related to beliefs that potentially motivate physicians in the design of surveillance strategies. When viewed independently of follow-up practice patterns, thoracic surgeons appear to be motivated by the desire to please patients, avoid malpractice suits, and improve patient quality of life. When viewed in relation to self-reported follow-up, none of these motivating factors were consistently associated with follow-up intensity. Belief in curative treatment of recurrence and enhanced likelihood of immediate palliative treatment leading to improved survival were the factors most frequently associated with variation in follow-up. Although the ability of the logistic and stepwise regression models to predict test use and follow-up intensity was less than optimal for TNM stage I patients, predictive ability was substantially improved for TNM stage II and III patients by including earlier-stage practice patterns as an independent variable. CONCLUSIONS: Physician characteristics and beliefs predicted a less than expected amount of the variation in self-reported follow-up intensity by TNM stage when modelled without knowledge of follow-up practice for any other TNM stage. Discrepancies between self-reported and actual follow-up may be partially responsible, although lack of surveillance guidelines is more likely. The inclusion of barriers to follow-up may improve future models. PMID- 9872181 TI - Prognosis of patients with lung cancer found in a single chest radiograph screening. AB - BACKGROUND: The prognosis of patients with lung cancer is better when the diagnosis is made early; the disease is localized, and radical surgery is possible. Screening for lung cancer with mass radiography or sputum cytology should contribute to a more favorable prognosis. Large-scale screening studies have improved the survival rates for lung cancer but have yielded no reduction in mortality rates. METHODS: The histologic types, stages, treatments, and survival rates were studied in 93 men who were found to have lung cancer in a single chest radiograph screening of more than 33,000 men who smoked and were 50 to 69 years old ("screened cases"), and in 239 men of the same age range whose lung cancer was detected through ordinary health care system ("other cases") during the screening period. RESULTS: The distribution of the histology was similar in the two groups, but screening detected more instances of early-stage disease that were resectable more often than in the other group (37 vs 19%). The 5-year survival rate for men in the screened cases was 19%, and that of men in the other cases was 10% (relative risk, 0.65; 95% confidence interval [CI], 0.50 to 0.84). The survival rate of men in the screened cases remained significantly higher than that of men in the other cases even after adjustments for age, smoking status, histology, stage of the disease, and resectability of the disease (relative risk, 0.74; 95% CI, 0.55 to 1.00). CONCLUSIONS: According to this study, chest radiograph screening might improve the prognosis of lung cancer. Our results are, however, subject to many factors that were only partially controlled for, and they should be interpreted cautiously. PMID- 9872183 TI - Digital clubbing and lung cancer. AB - OBJECTIVES: To determine the relative frequency of clubbing in small cell lung carcinoma (SCLC) versus non-small cell lung carcinoma (NSCLC). DESIGN: Examine patients with lung cancer for digital clubbing and relate the findings to the histopathologic subtype of lung cancer. SETTING: Cancer center at a tertiary teaching hospital. PATIENTS OR PARTICIPANTS: One hundred and eleven consecutive patients with a pathological diagnosis of lung cancer examined by one physician (KSS). INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Clubbing was present in 32 (29%) of the 111 patients with lung cancer. Clubbing was more common in women (40%) than in men (19%; chi2 test p = 0.011), and was more common in patients with NSCLC (35%) than those with SCLC (4%; chi2 test p = 0.0036). CONCLUSION: In a prospective study, digital clubbing was less frequently observed in men than women and in patients with SCLC than NSCLC. These clinical observations may assist in the initial evaluation of patients for planning workup and therapy. PMID- 9872184 TI - Prognostic significance of supraclavicular lymph nodes in small cell lung cancer: a study from four consecutive clinical trials, including 1,370 patients. "Petites Cellules" Group. AB - STUDY OBJECTIVE: To determine the prognostic significance of supraclavicular lymph node (SCLN) involvement in small cell lung cancer. MATERIALS AND METHODS: Patients (1,370) with small cell lung cancer were included in four consecutive clinical trials and classified as having either limited or extensive forms of disease using the Veterans Administration staging system. RESULTS: SCLN was present in 17% of patients (n = 234). Median survival was 258 days for patients with SCLN (n = 234) and 297 days for patients without SCLN (n = 1136) (p = 0.002). SCLN involvement was correlated with the presence of distant metastases at baseline (169 vs 65, p = < 0.001). Median survival was 375 days for patients with limited forms without SCLN (n = 529), 332 days for those with limited forms with SCLN (n = 65) (p = 0.12), 244 days for those with extensive forms without SCLN (n = 604), and 228 days for those with extensive forms with SCLN (n = 169) (p = 0.94). The 2-year survival rates were 17%, 12%, 2%, and 4%, respectively. Cox models confirmed that SCLN did not provide any significant additional prognostic information. CONCLUSION: SCLN is highly correlated with extensive forms explaining its overall prognostic value. In limited disease, SCLN is only a minor poor prognostic factor, not justifying any amendment to the staging system currently used. PMID- 9872185 TI - Sialyl stage-specific embryonic antigen-1: a useful marker for differentiating the etiology of pleural effusion. AB - OBJECTIVES: To assess the usefulness of sialyl stage-specific embryonic antigen-1 (SSEA-1) levels in differentiating the etiology of pleural effusion (PE). DESIGN: A solid-phase immunoradiometric sandwich assay with an FH6 monoclonal antibody was used to measure sialyl SSEA-1 levels in PEs of 132 patients with various diseases. Paired serum sialyl SSEA-1 levels were measured simultaneously in 47 patients with various subtypes of lung cancer RESULTS: The pleural sialyl SSEA-1 levels were significantly higher in patients who had adenocarcinoma of the lung with positive cytology than in all the other patients, including those having malignancies other than adenocarcinoma of the lung, adenocarcinoma of the lung with cytology-negative PE, and benign diseases. There were no significant differences among sialyl SSEA-1 levels in the pleural fluid containing no adenocarcinoma cells. Using the cutoff value of 265 U/mL, the sensitivity was 64% (25/39) and the specificity was 95% (88/93) for the pleural sialyl SSEA-1 level to differentiate adenocarcinoma from other effusions. CONCLUSIONS: With high specificity and modest sensitivity, the pleural sialyl SSEA-1 level is a useful biochemical marker for differentiating the etiology of PEs caused by adenocarcinoma from other diseases. PMID- 9872186 TI - Prospective evaluation of anticoagulant reversal with oral vitamin K1 while continuing warfarin therapy unchanged. AB - STUDY OBJECTIVES: To study the efficacy and safety of partially correcting therapeutic anticoagulation by administering oral vitamin K1. DESIGN: Prospective interventional trial. SETTING: Outpatient anticoagulation clinic. PATIENTS: Patients who required reversal of their normal or excessive therapy with oral anticoagulant drugs were recruited. INTERVENTIONS: All patients received a single oral dose of vitamin K1. The dose was calculated using a regression formula and was intended to decrease the international normalized ratio (INR) to a predetermined value. Patient follow-up continued for 8 weeks. We compared the actual change of the INR to the predicted change. RESULTS: Sixty-five reversals of anticoagulant therapy were initiated in the study group. Sixty-four of the 65 reversals were successful. The mean (+/-SEM) initial INR was 2.6+/-0.1 for the preprocedure patients and 8.4+/-0.5 for the excessively anticoagulated patients. The predicted change in the INR correlated with the actual change (r = 0.92, p < 0.0001). There were no thromboembolic events and only one hemorrhagic complication. The mean (+/-SEM) dose of oral vitamin K1 was 5.0+/-0.3 mg for the preprocedure patients and 10.0+/-1.0 mg for the excessively anticoagulated patients. CONCLUSIONS: The administration of a single oral dose of vitamin K1 is a safe and effective method for partially reversing anticoagulant therapy without disrupting the daily maintenance dose of warfarin. A reliable regression formula was developed to predict the dose of vitamin K1 needed to achieve the desired INR. PMID- 9872187 TI - Pulmonary embolism; lung scanning interpretation: about words. AB - OBJECTIVE: To assess clinicians' agreement on how they interpret lung scan reports with regard to the diagnosis of pulmonary embolism. DESIGN: In this prospective study, nuclear medicine physicians provided two types of reports for each lung scan: a routine descriptive report and a short form with a standardized conclusion on the likelihood of pulmonary embolism: "high probability," "no conclusion," and "diagnosis excluded." Three independent blinded senior clinicians reviewed all routine reports and chose one of the following conclusions: "high probability," "no conclusion," or "diagnosis excluded." SETTING: An acute care teaching hospital near Paris. SUBJECTS: Eighty-two lung scans were studied. MAIN OUTCOME MEASUREMENTS: Inter-clinician agreement and agreement between clinicians' conclusions and the nuclear medicine physicians' standardized reports were analyzed using the kappa index. RESULTS: The distribution of the clinicians' conclusions from routine reports strongly differed (p < 0.001). Agreement among the three clinicians was observed in 40.2% of the routine reports, and the inter-clinician agreement was poor to moderate (kappa range, 0.28 to 0.52). A complete agreement among the three clinicians and the nuclear medicine physicians' standardized conclusions was observed for 32.9% of the reports. The agreement between each clinician and the standardized conclusions was also poor to moderate (kappa range, 0.32 to 0.55). CONCLUSION: Reading the same routine reports, clinicians reached different conclusions. Furthermore these differed greatly from the nuclear medicine physicians' standardized conclusions. These results support the notion that physicians should be given standardized reports. PMID- 9872188 TI - Estrogen acutely abolishes abnormal cold-induced coronary constriction in men. AB - BACKGROUND AND STUDY OBJECTIVE: Ambient cold exposure may induce myocardial ischemia by precipitating coronary artery constriction and a decrease in coronary blood flow. Estrogen has vasoactive properties that may prevent abnormal coronary constriction in a sex-independent manner. The purpose of this study is to determine whether estrogen acutely abolishes abnormal coronary responses to cold exposure in men. DESIGN: Randomized, double-blinded placebo-controlled clinical trial. SETTING: Cardiac catheterization laboratory. PATIENTS: Men referred for routine diagnostic coronary angiography who exhibit abnormal coronary artery constriction in response to a 90-s cold pressor test (CPT). INTERVENTION: Intravenous conjugated estrogens (1.25 mg) vs. placebo. MEASUREMENTS AND RESULTS: Rate-pressure product, coronary cross-sectional area (CSA), and coronary blood flow responses to the CPT were measured before and 15 min after intervention. In 12 men with CPT-induced coronary constriction who were assigned to estrogen, CPT induced a mean 21.8% decrease in coronary CSA (p < 0.01) and a nonsignificant change in coronary flow. After estrogen, the repeated CPT induced a 16.3% increase in CSA (p < 0.01) and a 54.9% increase in flow (p < 0.01). CSA and coronary flow responses to CPT were significantly different before and after estrogen (p < 0.01). In contrast, placebo was not associated with changes in CSA or coronary flow responses to CPT in eight men. CONCLUSIONS: In men, conjugated estrogens acutely abolish abnormal coronary constriction and improve coronary blood flow responses to an exogenous cold stimulus. These results suggest that estrogen favorably alters coronary vasoreactivity in men. PMID- 9872189 TI - Exercise tolerance in asymptomatic elderly men with fluoroscopically detected coronary artery calcification. AB - STUDY OBJECTIVE: The value of detecting coronary artery calcification (CAC), by cardiac imaging, for the diagnosis of coronary artery disease (CAD) in asymptomatic middle-aged men has been demonstrated. However, the incidence of CAC increases with age. The functional significance of CAC remains unknown in asymptomatic elderly men. The purpose of this study is to explore whether CAC in asymptomatic aging men signifies the presence of cardiovascular dysfunction during exercise. DESIGN: This study was designed to address whether elderly asymptomatic men, selected because they have CAC, have reduced exercise tolerance due to functionally significant CAD. PARTICIPANTS AND SETTING: Thirty-eight asymptomatic male volunteers (ages 50 to 75 years, mean [+/-SD] 64+/-7 years) with a normal resting ECG and at least one coronary risk factor, in a population study. Nineteen subjects had CAC detected by digital subtraction fluoroscopy in at least two major coronary arteries, and 19 subjects had no identifiable CAC. METHODS AND RESULTS: Each subject underwent a symptom-limited incremental exercise test with 12-lead ECG monitoring and respiratory gas analysis. Four indexes of exercise oxygen transport were evaluated: peak oxygen uptake (VO2), lactic acidosis threshold, peak VO2/heart rate ratio, and VO2 relative to a work rate increase. Eleven of 38 subjects (28%) were found to have reduced oxygen transport, which was defined as an abnormal reduction in more than two of the above four indexes of oxygen transport. Five of the 11 subjects with reduced oxygen transport had CAC, and 6 subjects did not (not significant). Only one subject with CAC had exercise ST depression. CONCLUSION: Significant CAC in asymptomatic men over age 50 does not signify exercise limitation due to CAD. PMID- 9872190 TI - PREPARED: Preparation for Angiography in Renal Dysfunction: a randomized trial of inpatient vs outpatient hydration protocols for cardiac catheterization in mild to-moderate renal dysfunction. AB - BACKGROUND: IV hydration before and after cardiac catheterization is effective in preventing contrast-associated renal dysfunction for patients with mild-to moderate renal insufficiency, but necessitates overnight hospital admission. We tested an outpatient oral precatheterization hydration strategy in comparison with overnight IV hydration. METHODS: We randomized 36 patients with renal dysfunction (serum creatinine > or = 1.4 mg/dL) undergoing elective cardiac catheterization to receive either overnight IV hydration (0.45 normal saline solution at 75 mL/h for both 12 h precatheterization and postcatheterization; n = 18) or an outpatient hydration protocol including precatheterization oral hydration (1,000 mL clear liquid over 10 h) followed by 6 h of IV hydration (0.45 normal saline solution at 300 mL/h) beginning just before contrast exposure. The predefined primary end point was the maximal change in creatinine up to 48 h after cardiac catheterization. RESULTS: The inpatient and outpatient groups were well matched for baseline characteristics and contrast volume. By protocol design, the outpatient group received a greater volume of hydration, although the net volume changes were comparable in the two groups. The maximal changes in serum creatinine in the inpatient (0.21+/-0.38 mg/dL; 95% confidence interval [CI], 0.02 to 0.39 mg/dL) and outpatient groups (0.12+/-0.23 mg/dL; 95% CI, 0.01 to 0.24 mg/dL) were comparable (p = not significant). There were no instances of protocol intolerance. CONCLUSIONS: A hydration strategy compatible with outpatient cardiac catheterization is comparable to precatheterization and postcatheterization IV hydration in preventing contrast-associated changes in serum creatinine. Hospital admission for IV hydration is unnecessary before elective cardiac catheterization in the setting of mild-to-moderate renal dysfunction. PMID- 9872191 TI - Effects of lung volume reduction surgery on exercise pulmonary hemodynamics in severe emphysema. AB - STUDY OBJECTIVES: To clarify the effects of lung volume reduction surgery (LVRS) on pulmonary hemodynamics in severe emphysema and to evaluate the role of pulmonary circulation in the increased exercise performance after LVRS. DESIGN: In eight male patients with severe emphysema, we measured pulmonary artery (Ppa) and occlusion (Pop) pressures and cardiac output through a Swan-Ganz thermodilution catheter, and we calculated cardiac index (CI), pulmonary vascular resistance index, and driving pressure. The study was performed at rest and during exercise using a supine bicycle ergometer at 25 W, under room air and O2 inhalation. Exercise performance was expressed as endurance time in minutes. Pulmonary function tests were performed. The patients underwent an identical study before and 6 months after LVRS. RESULTS: The patients' exercise performance was significantly increased after LVRS (5.9+/-1.8 min) compared with that before LVRS (3.6+/-0.9 min). LVRS resulted in a significant increase in FEV1 and a significant decrease in residual volume. Before LVRS, Ppa was increased both at rest and during exercise. LVRS did not change rest or exercise Ppa. LVRS significantly decreased Pop during exercise from 24+/-10 mm Hg to 18+/-7 mm Hg. CI at rest and during exercise were significantly increased after LVRS. O2 administration significantly decreased Ppa during exercise both before and after LVRS. CONCLUSIONS: These findings suggest that LVRS does not improve pulmonary hypertension at rest or during exercise in patients with severe emphysema and that elevated Pop during exercise before LVRS is probably related to lung mechanic abnormalities. PMID- 9872192 TI - Estimated growth of lung volume reduction surgery among Medicare enrollees: 1994 to 1996. AB - OBJECTIVE: To estimate the number of lung volume reduction surgery procedures performed on Medicare enrollees from 1994 to 1996. DESIGN: Statistical analysis of national Medicare claims data. PATIENTS: All Medicare enrollees with emphysema hating claims records for pulmonary resection procedures from January 1, 1993, through December 31, 1996. MAIN OUTCOME MEASURE: Estimated number of lung volume reduction procedures performed per month from July 1994 through December 1996. RESULTS: An estimated 1,212 lung volume reduction procedures were performed on Medicare enrollees between July 1994 and December 1995 (95% confidence interval, 1,012 to 1,408). Nearly one half of these procedures were performed in the last 3 months of 1995. At the time Health Care Financing Administration announced that it would suspend reimbursement for the procedure (December 1995), lung volume reduction surgery was being performed in 37 states. The number of claims per month decreased from a peak of 169 in December 1995, to 11 in March 1996. Average Medicare reimbursement per procedure was $31,398. CONCLUSIONS: Lung volume reduction surgery for patients increased rapidly following its reintroduction in 1994. The growth of lung volume reduction surgery demonstrates that widespread adoption and utilization of a surgical procedure can occur in the absence of data from controlled clinical trials. Medicare expenditures for lung volume reduction surgery were an estimated $30 million to $50 million. Performing the surgery for all current Medicare patients who meet the appropriate clinical criteria would cost an estimated $1 billion. PMID- 9872193 TI - Etiology of community-acquired pneumonia in hospitalized patients: a 3-year prospective study in Japan. AB - STUDY OBJECTIVE: To compare the etiology of community-acquired pneumonia in Japan and Western countries, the causative pathogens were prospectively investigated in patients requiring hospitalization. DESIGN: Prospective study over a 3-year period. SETTING: A community general hospital in Japan. PATIENTS: Three hundred twenty-six episodes of community-acquired pneumonia in 318 patients admitted to the hospital between July 1994 and June 1997. METHODS: The microbiological diagnosis was based on the results of quantitative sputum culture, blood culture, and other invasive procedures, including transthoracic needle aspiration or bronchoscopic examination. Serologic tests for Mycoplasma pneumoniae, Chlamydia spp, Legionella spp, and viruses were also routinely performed. RESULTS: Causative pathogens were identified in 199 episodes (61%). Streptococcus pneumoniae was the most common pathogen (23%), followed by Haemophilus influenzae (7.4%), M pneumoniae (4.9%), and Klebsiella pneumoniae (4.3%). The Streptococcus milleri group and Chlamydia pneumoniae were detected in 3.7 and 3.4% of the episodes, respectively. Pneumonia due to Legionella spp was recognized in only two patients. CONCLUSIONS: The etiology of community-acquired pneumonia in Japan did not differ markedly when compared with that of Western countries except for the low incidence of Legionella pneumonia. C pneumoniae and the S milleri group, which are emerging or newly recognized pathogens, were also significant causative microorganisms. PMID- 9872194 TI - The effect of Pseudomonas aeruginosa infection on clinical parameters in steady state bronchiectasis. AB - STUDY OBJECTIVE: To investigate the effect of Pseudomonas aeruginosa infection on clinical parameters in Chinese patients with noncystic fibrosis and steady-state bronchiectasis. DESIGN: Prospective, cross-sectional clinicomicrobiological study with informed consent. SETTING: Consecutive outpatient recruitment from a specialist bronchiectasis respiratory clinic. PATIENTS: Outpatients (n = 100; 62 women; 55.1+/-16.7 years old; FEV1/FVC 1.4+/-0.7/2.1+/-0.9 L), who had stable respiratory symptoms for more than 3 weeks. MEASUREMENTS AND RESULTS: Respiratory pathogens isolated from the sputum were: Pseudomonas aeruginosa (33), Haemophilus influenzae (10), Moraxella catarrhalis (2), other Gram-negative bacilli (5), Streptococcus pneumoniae (6), Staphylococcus aureus (5), mycobacteria (3), and yeast (1). Clinical parameters in patients with positive isolation of P aeruginosa were compared with those without the organism in the sputum culture (non-P aeruginosa). In the P aeruginosa group, the FEV1/FVC ratio and sputum volume were lower (p < 0.005) and higher (p < 0.0001), respectively, than those of the non-P aeruginosa group. The FEV1/FVC ratio (< 60%) and sputum volume (grading > 5) were independently associated with a positive sputum isolation of P aeruginosa with odds ratios of 3.1 (confidence interval [CI] 1.2 to 8.4; p < 0.01) and 4.7 (CI 1.6 to 13.3; p < 0.001), respectively. CONCLUSIONS: P aeruginosa is the predominant respiratory pathogen isolated in the sputum of Chinese patients with steady-state bronchiectasis, and its isolation is associated with high sputum output (> or = 75th quartile) and moderately severe airflow obstruction (FEV1/FVC < 60%). PMID- 9872195 TI - Factors related to the relapse of bronchiolitis obliterans organizing pneumonia. AB - STUDY OBJECTIVE: The purpose of this study is to determine factors, including laboratory data, related to the relapse of bronchiolitis obliterans organizing pneumonia (BOOP). DESIGN: Retrospective study. SETTING AND PATIENTS: The medical files of Fukuoka University Hospital and Nishi Fukuoka Hospital patients from 1984 to 1996 were reviewed, and 18 cases of BOOP that had been diagnosed using transbronchial or open lung biopsy were selected for evaluation. MEASUREMENTS: The 18 cases were put into two groups composed of 7 patients who relapsed and 11 who did not relapse. Their clinical symptoms and laboratory data at first admission, including hemograms, blood chemistry tests, and pulmonary function tests were compared. Patients with or without associated diseases, such as collagen vascular diseases, were compared using the same parameters in order to examine the relationship between the associated diseases and BOOP relapse. RESULTS: The serum levels of total protein and albumin in patients who relapsed were significantly lower than in patients who did not relapse, respectively: 5.8 (range, 4.4 to 6.2) vs 6.3 (range, 4.5 to 6.8) g/dL, p < 0.05; and 2.9 (range, 2.5 to 3.4) vs 3.7 (range, 2.8 to 4.3) g/dL, p < 0.01. Levels of serum albumin in BOOP patients with associated diseases, however, were significantly lower than in those without associated diseases, respectively: 2.95 (range, 2.5 to 3.9) vs 3.65 (range, 2.8 to 4.3) mg/dL, p < 0.05. The fall in serum albumin levels in patients who relapsed, therefore, was probably due to associated diseases. The fact that 5 of 8 patients with associated diseases relapsed but only 2 of 10 without associated diseases relapsed suggests that a relationship exists between associated diseases and the prognosis of BOOP, although this finding was not statistically significant because of the small number of cases and the heterogeneity of the associated diseases. The most striking observation was that PaO2 levels in patients who relapsed were significantly lower than in those who did not, respectively: 55.4 (range, 39.9 to 73.2) vs 78.0 (range, 48.4 to 89.4) mm Hg, p < 0.05. However, PaO2 levels were not statistically different between patients with and without associated diseases, respectively: 66.0 (range, 45.4 to 78.8) vs 71.4 (range, 39.9 to 89.4) mm Hg. CONCLUSIONS: The severity of hypoxemia at first medical examination may be an important determinant for the subsequent BOOP relapse. PMID- 9872196 TI - Assessing the reversibility of airway obstruction. AB - STUDY OBJECTIVE: To determine whether changes of partial expiratory flow-volume curve (PEFV) and inspiratory capacity (IC) detect functional responses to bronchodilator in patients who do not meet the FEV1 criteria for reversibility of airway obstruction. DESIGN/METHODS: The effects of salbutamol (200 microg by metered-dose inhaler) on lung function were examined in 50 patients with asthma and 28 patients with COPD. Measurements evaluated were FEV1, forced expiratory flow at 30% of control FVC from maximal expiratory flow-volume curve (Vm30), forced expiratory flow at 30% of control FVC from PEFV (Vp30), and IC. On a separate occasion, a representative sample of 26 subjects inhaled placebo to determine the 95% confidence limits (CLs) of each of the parameters. RESULTS: A percent and absolute increment of FEV1 above the upper CL was recorded in 28 patients. Of these, 26 had a percent and absolute increase of Vp30, 21 of Vm30, 9 of FVC, and 11 of IC above the 95% CL. Of the 50 patients who did not have an increase in FEV1 above the 95% CL, 25 had a percent and absolute increase in Vp30, 15 of Vm30, 3 of FVC, and 13 of IC above the 95% CL. On average, the percent and absolute increase Vp30 above the 95% CL significantly identified more responders than every other parameter. CONCLUSION: Increases in maximal flow detected by PEFV and/or changes in IC may be substantially obscured by the effects of inspiration to total lung capacity required for the measurement of FEV1 in patients with chronic bronchoconstriction. Decreases in functional residual capacity (FRC) manifested by an increase of IC occur because, in patients whose FRC is dynamically determined, bronchodilatation that increases maximal flow in the tidal breathing range allows patients to breathe at lower lung volumes. Changes of FEV1 frequently fail to detect significant functional response to bronchodilators in patients with chronic airflow obstruction. PMID- 9872197 TI - Exercise-induced bronchospasm in high school athletes via a free running test: incidence and epidemiology. AB - BACKGROUND: Exercise-induced bronchospasm (EIB) affects up to 35% of athletes and up to 90% of asthmatics. Asthma morbidity and mortality have increased over the past several decades among residents of Philadelphia, PA. It is possible that a simple free running test for EIB may serve as a tool to study the factors contributing to recent trends in asthma, and to screen for asthma in athletes in the urban setting. OBJECTIVES: The purposes of this study were to (1) assess a free running test to screen for EIB, and (2) examine prevalence of and epidemiologic factors associated with EIB in high school athletes. DESIGN: Cross sectional observational study on the incidence and risk factors for EIB. To validate our method and criteria for the diagnosis of EIB, a repeat test was performed on a portion of the athletes. In a randomized single-blinded fashion, 15 athletes who had demonstrated EIB initially received albuterol or placebo prior to a repeat exercise test. SETTING: Community high school athletic facilities. PARTICIPANTS: We studied 238 male high school varsity football players. INTERVENTION: All athletes underwent an acquaintance session with a questionnaire, followed by a 1-mile outdoor run (6 to 8 mins). MEASUREMENTS: Peak expiratory flow (PEF) measurements were determined prior to and 5, 15, and 30 min after exercise. Heart rates (HRs) and dyspnea scores were measured. EIB was defined as a decrease of 15% in PEF at any time point after exercise. Associations of EIB with demographic factors were assessed by univariate and multivariate analyses. RESULTS: Two hundred thirty-eight athletes participated: 92 European-Americans (EA), 140 African-Americans (AA), 5 Hispanics, and 1 Native American. Mean age was 16+/-1 years. Average HR postexercise was 156+/-24 beats/min. Twenty-four (10%) reported a history of treated asthma. The prevalence of EIB among the remaining 214 athletes was 19 of 214 (9%). The rate of EIB among AA athletes was higher than among EA athletes: (17/126 [13%] AA vs 2/82 [2%] EA, p = 0.01). During the validation portion of the study, the placebo-treated group (n = 7) demonstrated a consistent drop in PEF after exercise on repeat testing, with a 16+/-5% fall in PEF on initial testing and a 14+/-13 drop with placebo. In contrast, the fall in airflow in the albuterol-treated athletes (n = 8) following exercise reversed with albuterol treatment, from a 15+/-6% fall in PEF at initial testing to an increase in PEF of 6+/-9% from baseline following albuterol administration. A history of wheezing (p < 0.001), residence in a poverty area (p < 0.0001), race (p = 0.01), remote history of asthma (p < 0.001), and absolute water content of the air on the day tested (p = 0.04) were significantly associated with EIB. By stepwise regression, EIB was most closely associated with a history of wheezing (p = 0.001) and poverty area residence (p = 0.003). CONCLUSIONS: Our findings indicate a substantial rate of unrecognized EIB exists among urban varsity athletes, and suggest that active screening for EIB, especially for students residing in poverty areas, may be indicated to identify individuals at risk for EIB and asthma. PMID- 9872198 TI - Relationship between abnormalities on high-resolution CT and pulmonary function in systemic sclerosis. AB - STUDY OBJECTIVES: To determine the predictive value of abnormalities on high resolution CT (HRCT) on pulmonary disease in systemic sclerosis. PATIENTS: Fifty two patients suffering from systemic sclerosis. DESIGN: Pulmonary disease was defined by pulmonary function test abnormalities, ie, total lung capacity (TLC) <80% of predicted value and/or diffusion of carbon monoxide (DLCO) <75% of predicted value, without any pulmonary event other than systemic sclerosis in the medical history. Patients were divided in two groups, group A with pulmonary disease (29 patients) and group B without pulmonary disease (23 patients). HRCT abnormalities were scored on whole lungs. A decision matrix was constructed to determine sensitivity, specificity, positive and negative predictive values, and false-positive and false-negative rates. A receiver operating characteristic curve was constructed to determine the best compromise between sensitivity and specificity. RESULTS: HRCT total scores were higher in group A (9.0+/-4.3) than in group B (5.0+/-2.8) (p < 0.001) and they correlated with TLC (r =-0.39, p < 0.005) and DLCO (r = -0.50, p < 0.0002). An HRCT score of 7 corresponded to the best compromise between sensitivity (0.60) and specificity (0.83), with a positive predictive value of 0.82. Taking into account a value of 10 for the HRCT score increased specificity to 1 but decreased sensitivity to 0.41. CONCLUSION: A minimum score of 7 would be required to consider HRCT abnormalities in systemic sclerosis as predictive of pulmonary disease. An HRCT score of 10 makes it possible to establish the diagnosis of lung involvement severe enough to impair pulmonary function. PMID- 9872199 TI - Treatment success with a mandibular advancement device is related to supine dependent sleep apnea. AB - STUDY OBJECTIVE: To evaluate the effect of a mandibular advancement device in patients with supine-dependent sleep apnea and patients with non-supine-dependent sleep apnea. DESIGN: Prospective study. SETTING: Department of Respiratory Medicine, University Hospital, Umea, Sweden. PATIENTS: Twenty-six patients with obstructive sleep apnea. INTERVENTION: Individually fabricated and adjusted mandibular advancement devices. MEASUREMENTS: Overnight polysomnographic sleep recordings with and without the device. Supine-dependent sleep apnea was defined when the supine apnea-hypopnea index was > or = 10, together with a lateral apnea hypopnea index of < 10. Non-supine-dependent sleep apnea was considered when the lateral apnea-hypopnea index was > or = 10. RESULTS: In 12 patients with supine dependent sleep apnea, the device reduced the supine apnea-hypopnea index from a median of 41 (range, 16 to 70) to 5.9 (range, 0.0 to 15) (p < 0.01). In 14 patients with non-supine-dependent sleep apnea, the treatment reduced the supine apnea-hypopnea index from 44 (range, 1.8 to 73) to 21 (range, 6.3 to 60) (p < 0.05) and the lateral apnea-hypopnea index from 21 (range, 12 to 70) to 4.5 (range, 0.0 to 31) (p < 0.01). The odds ratio for a successful apnea reduction to an apnea-hypopnea index of < 10 in both the supine and the lateral positions was 30 for supine-dependent sleep apnea adjusted for age, obesity, mandibular advancement, and mandibular opening (p < 0.01). CONCLUSION: Successful apnea reduction with a mandibular advancement device is highly related to supine dependent sleep apnea. PMID- 9872200 TI - Comparison of noninvasive positive pressure ventilation with standard medical therapy in hypercapnic acute respiratory failure. AB - STUDY OBJECTIVE: To compare the efficacy of standard medical therapy (ST) and noninvasive mechanical ventilation additional to standard medical therapy in hypercapnic acute respiratory failure (HARF). DESIGN: Single center, prospective, randomized, controlled study. SETTING: Pulmonary medicine directed critical care unit in a university hospital. PATIENTS: Between March 1993 and November 1996, 30 HARF patients were randomized to receive ST or noninvasive positive pressure ventilation (NPPV) in addition to ST. INTERVENTIONS: NPPV was given with an air cushioned face via a mechanical ventilator (Puritan Bennett 7200) with initial setting of 5 cm H2O continuous positive airway pressure and 15 cm H2O pressure support. RESULTS: At the time of randomization, patients in the ST group had (mean+/-SD) PaO2 of 54+/-13 mm Hg, PaCO2 of 67+/-11 mm Hg, pH of 7.28+/-0.02, and respiratory rate of 35.0+/-5.8 breaths/min. Patients in the NPPV group had PaO2 of 55+/-14, PaCO2 of 69+/-15, pH of 7.27+/-0.07, and respiratory rate of 34.0+/ 8.1 breaths/min. With ST, there was significant improvement of only respiratory rate (p < 0.05). However, with NPPV, PaO2 (p < 0.001), PaCO2 (p < 0.001), pH (p < 0.001), and respiratory rate (p < 0.001) improved significantly compared with baseline. Six hours after randomization, pH (p < 0.01) and respiratory rate (p < 0.01) in NPPV patients were significantly better than with ST. Hospital stay for NPPV vs ST patients was, respectively, 11.7+/-3.5 and 14.6+/-4.7 days (p < 0.05). One patient in the NPPV group required invasive mechanical ventilation. The conditions of six patients in the ST group deteriorated and they were switched to NPPV; this was successful in four patients, two failures were invasively ventilated. CONCLUSION: This study suggests that early application of NPPV in HARF patients facilitates improvement, decreases need for invasive mechanical ventilation, and decreases the duration of hospitalization. PMID- 9872203 TI - Fiberoptic bronchoscopy in coronary care unit patients: indications, safety, and clinical implications. AB - STUDY OBJECTIVES: To evaluate the indications, safety, therapeutic impact, and outcome of fiberoptic bronchoscopy (FOB) in coronary care unit (CCU) patients. DESIGN: Retrospective review of all CCU patients undergoing FOB during a 6-year period. SETTING: Tertiary care university hospital. RESULTS: Among 8,330 patients admitted to the CCU; 40 (0.5%) patients underwent FOB to evaluate pulmonary abnormalities, most often (78%) to appraise clinically suspected pneumonia. Thirty-five (88%) patients were intubated and 21 (53%) had acute myocardial infarction (MI) before FOB. There were two major complications (bleeding, intubation) occurring within 24 h of FOB, one of which appeared due to the procedure. No episodes of chest pain or ischemic events were recorded and no significant increase in major complications was noted in MI patients (3% vs 5%). Patients having FOB within 10 days of MI had higher survival (79%) than those undergoing FOB later (29%) (p = 0.05). Seven different bacterial pathogens were isolated in 6 (15%) patients, probably reflecting prior empiric antibiotics in 32 (80%) patients. Therapy was changed in 64% of patients in whom a potential pathogen was identified. Despite alterations in treatment, patients with clinically suspected pneumonia and any organisms isolated by FOB had greater mortality (79% vs 31%, p = 0.003) than those with sterile FOB cultures. CONCLUSION: FOB may be diagnostically useful in the evaluation of pulmonary abnormalities in selected patients with acute cardiac disease, can be performed safely, and may influence management decisions. Positive bronchoscopy cultures often influence therapy but are associated with higher mortality, suggesting a lethal effect of nosocomial pneumonia in this subset of CCU patients. The risks of FOB must be weighed with the impact of FOB results on patient outcome, and its role requires further investigation. PMID- 9872201 TI - Multicenter study of noninvasive monitoring systems as alternatives to invasive monitoring of acutely ill emergency patients. AB - BACKGROUND: Recent reports showed lack of effectiveness of pulmonary artery catheterization in critically ill medical patients and relatively late-stage surgical patients with organ failure. Since invasive monitoring requires critical care environments, the early hemodynamic patterns may have been missed. Ideally, early noninvasive hemodynamic monitoring systems, if reliable, could be used as the "front end" of invasive monitoring to supply more complete descriptions of circulatory pathophysiology. OBJECTIVES: To evaluate the accuracy and reliability of noninvasive hemodynamic monitoring consisting of a new bioimpedance method for estimating cardiac output combined with arterial BP, pulse oximetry, and transcutaneous PO2 and PCO2; we compared this system of noninvasive monitoring with simultaneous invasive measurements to evaluate circulatory deficiencies in acutely ill patients shortly after hospital admission where invasive monitoring was not readily available. We also preliminarily explored early differences in temporal hemodynamic patterns of survivors and nonsurvivors. DESIGN AND SETTING: Prospective comparison of simultaneous invasive and noninvasive measurements of circulatory function with retrospective analysis of data in university-run county hospitals, university hospitals and affiliated teaching hospitals, and a community private hospital. PATIENTS: We studied 680 patients, including 139 severely injured or hemorrhaging patients in the emergency department (ED), 129 medical (nontrauma) patients on admission to the ED, 274 high-risk surgical patients intraoperatively, and 138 patients recently admitted to the ICU. RESULTS: A new noninvasive impedance device provided cardiac output estimations under conditions in which invasive thermodilution measurements were not usually applied. There were 2,192 simultaneous bioimpedance and thermodilution cardiac index measurements; the correlation coefficient, r = 0.85, r2 = 0.73, p < 0.001. The precision and bias was -0.124+/-0.75 L/min/m2. Both invasive and noninvasive monitoring systems provide similar information and identified episodes of hypotension, low cardiac index, arterial hemoglobin desaturation, low transcutaneous O2, high transcutaneous CO2, and low oxygen consumption before and during initial resuscitation. The limitations of noninvasive systems were described. CONCLUSIONS: Noninvasive monitoring systems gave continuous displays of physiologic data that provided information allowing early recognition of low flow and poor tissue perfusion that were more pronounced in the nonsurvivors. Noninvasive systems may be acceptable alternatives where invasive monitoring is not available. PMID- 9872202 TI - Exhaled breath condensate isoprostanes are elevated in patients with acute lung injury or ARDS. AB - BACKGROUND: Oxidant stress is a purported mechanism of tissue damage in patients with ARDS and acute lung injury (ALI). Isoprostanes, prostanoid compounds primarily formed nonenzymatically via lipid peroxidation, are precise markers of in vivo oxidant stress. Plasma levels of metabolites of 8-iso-prostaglandin F2alpha (8-iso-PGF2alpha) correlate with outcome in patients with ARDS. OBJECTIVE: To examine exhaled breath condensate levels of 8-iso-PGF2alpha as a noninvasive quantification of pulmonary oxidant stress in patients with, or at risk for, ARDS/ALI. METHODS: Breath condensate was collected from 22 patients with, or at risk for, ARDS/ALI by placing Tygon tubing submerged in an ice bath in line with the expiratory limb of the ventilator circuit. Ten patients without lung disease, who were intubated while undergoing minor surgical procedures, served as control subjects. Between 1 and 3 mL of condensate was collected over a 30- to 60-min period, then immediately frozen and stored at -70 degrees C until analysis. The 8-iso-PGF2alpha was purified and derivatized, then quantified by stable isotope dilution in conjunction with gas chromatography/mass spectrometry. RESULTS: The mean level of exhaled 8-iso-PGF2alpha in the patients with ALI/ARDS, 87+/-28 pg/mL, was significantly higher than the mean in the normal group, 7+/-4 pg/mL (p = 0.007). The 8-iso-PGF2alpha levels were greater than two standard deviations above the mean of the normal group in 12 of 22 patients with or at risk for ARDS/ALI. CONCLUSIONS: These results provide further evidence that lipid peroxidation does occur in patients with ARDS/ALI. The measurement of exhaled isoprostanes provides a novel, noninvasive method to quantify oxidant stress in such patients. PMID- 9872204 TI - Vascular endothelial growth factor increased by pulmonary surgery accelerates the growth of micrometastases in metastatic lung cancer. AB - BACKGROUND: The use of surgery for metastatic lung cancer has been established recently and the indications have been extended to multiple and bilateral lung metastases. However, in some patients, secondary lung metastasis appears soon after the first pulmonary surgery, making curative treatment very difficult. Postoperative weakness of tumor angiogenesis suppression mechanisms seems to play an important role in the recurrence of lung metastases. To verify this hypothesis, we performed a clinical and an experimental study. RESULTS AND CONCLUSION: The clinical study revealed that serum vascular endothelial growth factor (VEGF), also known as vascular permeability factor, increased after pulmonary surgery. The experimental study showed that VEGF played an important role in the rapid growth of dormant micrometastases of the lung. These results suggested that the postoperative increase of VEGF disrupted angiogenesis suppression and induced the growth of dormant micrometastases early in the postoperative period. It was also demonstrated that this effect of VEGF on micrometastases was abolished by AGM-1470, an angiogenesis inhibitor. In conclusion, postoperative treatment with AGM-1470 might inhibit the early recurrence of malignant tumors. PMID- 9872205 TI - In vitro comparison of beclomethasone and salbutamol metered-dose inhaler aerosols inhaled during pediatric tidal breathing from four valved holding chambers. AB - STUDY OBJECTIVES: To determine the amount of salbutamol or beclomethasone delivered from metered-dose inhalers (MDIs) in particle sizes appropriate for inhalational treatment with four common holding chambers (AeroChamber, OptiChamber, Space Chamber, E-Z Spacer) when used under simulated pediatric tidal breathing conditions. DESIGN: Five devices of each type were tested with salbutamol (100 microg, Glaxo-Wellcome) and beclomethasone (50 microg, Glaxo Wellcome) MDIs. Each device was connected to face replicas representative of 7 month-old (infant), 2-year-old (toddler), and 4-year-old (child) children and aerosol inhaled into a cascade impactor (Anderson) using a valve system and simulated tidal breathing patterns representative of children of these ages. MEASUREMENTS AND RESULTS: Amounts of drug inhaled in fine particles varied significantly between devices and ages (p < 0.01), eg, for beclomethasone, from 4 microg (infant, Space Chamber) to 8 microg (toddler, OptiChamber), and for salbutamol, from 18 to 36 microg. The AeroChamber and OptiChamber behaved quite similarly, delivering more drug in the fine particles than the other two devices, and having insignificant variations in these amounts with age (p > 0.05). CONCLUSIONS: Amounts of drug inhaled in fine particles during pediatric tidal breathing from valved holding chambers are dependent on the holding chamber model, the drug used, and the breathing pattern, although dependence on the breathing pattern is relatively small for some of the devices tested. PMID- 9872206 TI - Permissiveness of guinea pig alveolar macrophage subpopulations to acute respiratory syncytial virus infection in vitro. AB - BACKGROUND AND OBJECTIVES: Alveolar macrophages (AMs) are targets for respiratory syncytial virus (RSV) infection in vivo and in vitro. However, only a minority of AMs are permissive to acute RSV infection in vitro, and it is unknown whether this permissiveness may be related to the degree of cellular maturation that is achieved in vivo. METHODS: By using density gradient centrifugation, in which the degree of AM maturation is inversely related to buoyant density, we prepared three subpopulations of guinea pig AMs (designated as hypodense, intermediate density, and high-density AMs). Twenty-four hours after exposure to RSV in vitro, the percentage of RSV-positive cells in each subpopulation was determined by immunocytochemistry; intracellular virus was released from cells by sonication and quantified by plaque assay, and intracellular localization of RSV proteins was evaluated by immunogold electron microscopy. RESULTS: High-density AMs had a significantly higher proportion of RSV-positive cells than hypodense AMs (p < 0.001), with intermediate-density AMs having intermediate values. The amounts of intracellular virus significantly increased from hypodense to intermediate density to high-density AMs (p < 0.001). Hypodense cells showed immunogold labeling principally within phagolysosomes, whereas intermediate-density and high density cells showed immunolabeling of free cytoplasmic viral proteins and nucleocapsids. CONCLUSIONS: The permissiveness of guinea pig AMs to acute RSV infection in vitro is inversely related to their degree of maturation achieved in vivo. In addition, these results suggest that immature, high-density AMs support RSV replication whereas more mature, hypodense AMs may restrict viral replication. PMID- 9872207 TI - Lymphangioleiomyomatosis: a review. PMID- 9872208 TI - Geriatric respiratory medicine. PMID- 9872209 TI - The lung as an alternative route of delivery for insulin in controlling postprandial glucose levels in patients with diabetes. AB - STUDY OBJECTIVES: To determine the efficacy of the lung as an alternative route of delivery for insulin in controlling glucose below diabetic levels (11.2 mmol/L) 2 h after the ingestion of a meal in patients with type 2 diabetes mellitus. DESIGN: Single-blinded, nonrandomized, placebo-controlled pilot study consisting of two visits. SETTING: A primary care facility. PATIENTS: Seven patients with type 2 diabetes mellitus. INTERVENTIONS: On the first study visit, fasting glucose levels were normalized. Then, patients inhaled 1.5 U/kg insulin by aerosol into the lungs 5 min before ingesting a test meal. On the second visit, patients inhaled placebo aerosol 5 min before ingesting the same meal. On both visits, plasma samples were collected and analyzed for glucose levels for 3 h during the postprandial state. MEASUREMENTS AND RESULTS: No one coughed after inhalation of insulin aerosol or demonstrated hypoglycemia. During the postprandial period, glucose levels were significantly lower at 20 min (5.12+/ 1.08 mmol/L), 1 h (7.87+/-0.73 mmol/L), 2 h (8.05+/-1.24 mmol/L) and 3 h (7.50+/ 1.43 mmol/L) following inhalation of insulin than when the placebo was used. Data for the placebo were 10.36+/-1.23 mmol/L at 20 min, 14.0+/-1.68 mmol/L at 1 h, 16.18+/-1.45 mmol/L at 2 h, and 14.37+/-2.11 mmol/L at 3h (for all comparisons, p < 0.05). On the insulin visit, glucose levels were < 11.2 mmol/L 2 h after the meal in six of seven patients. None attained this level at the placebo visit. In addition, glucose levels were within the normal postprandial range of < 7.84 mmol/L in four of seven patients 2 h after eating on the insulin visit. CONCLUSIONS: These results suggest that, once plasma glucose levels are normalized, postprandial glucose levels can be maintained below diabetic levels by delivering 1.5 U/kg insulin into the lungs 5 min before the ingestion of a meal. PMID- 9872210 TI - Variability in the classification of radiographs using the 1980 International Labor Organization Classification for Pneumoconioses. AB - This study describes the extent of agreement in classification of chest radiographs using the International Labor Organization (ILO) classification among six readers from the United States and Canada. A set of 119 radiographs was created and read by three Canadian and three US readers. The two ratings of interest were profusion (scored from 0/- to 3/+) and pleural abnormalities consistent with pneumoconiosis (scored with the ILO system, then collapsed into a yes/no). We used a number of approaches to evaluate interreader agreement on profusion and pleural changes, determining concordance, observed agreement, kappa statistic, and a new measure to approximate sensitivity and specificity. This study found that five of six readers had good fair to good agreement for pleural findings and for profusion as a dichotomous variable (> or = 1/0 vs < or = 0/1) using the kappa statistic, while a sixth reader had poor agreement. We found that concordance, expressed as percent agreement, was higher for normal radiographs than for ones that showed disease, and describe the use of the kappa statistic to control for this finding. This analysis adds to the existing literature with the use of the kappa statistic, and by presenting a new measure for "underreading" and "overreading" tendencies. PMID- 9872211 TI - Preoperative diagnosis with video-assisted thoracoscopy with miniaturized endoscopes in general thoracic surgery: a preliminary study. AB - Video-assisted thoracoscopy using a miniaturized endoscope (mini-VAT) was applied for preoperative diagnosis in general thoracic surgery. Thirty-one patients, including 27 with indeterminate pulmonary nodule and 4 with suspected pleural involvement of lung cancer or metastatic pleural tumor, underwent mini-VAT. As a pilot study, 14 of the former 27 patients underwent mini-VAT while receiving general anesthesia. As a prospective study, all the remaining 17 patients underwent mini-VAT while receiving local anesthesia. Solid scopes of three different sizes, 0.9, 1.9, or 4.0 mm diameter, were used. An artificial pneumothorax for scope introduction was produced by needle thoracentesis under atmospheric pressure. Automatic cutting needle biopsy was used for tissue sampling. In the pilot study group, mini-VAT with a 4.0-mm scope provided excellent visibility and diagnostic sensitivity of 100%. This study group showed the diagnostic sensitivity of needle biopsy for pulmonary nodule to be 100%. Hemorrhages and air leaks at biopsy sites were sealed with blood coagulation in a short time. In the prospective study group, mini-VAT with a 4.0-mm scope with the patients receiving local anesthesia provided a diagnostic sensitivity of 91% for pulmonary nodule and a diagnostic accuracy of 100% for suspected pleural involvement. Causes of failure of mini-VAT with the use of local anesthesia were cough reflex during needle biopsy and incomplete lung collapse for deeply located target in two cases. The adverse effects of the mini-VAT were paradoxical respiration in two cases in which local anesthesia was used. The patients who received only local anesthesia required no chest tube drainage. Mini-VAT is a simple, minimally invasive procedure suitable as a preoperative examination technique for histologic diagnosis, evaluation of disease progression, and selection of strategy in thoracic surgery. PMID- 9872212 TI - Management of anticoagulants in a patient requiring major surgery. PMID- 9872213 TI - A patient with acute exacerbation of COPD who did not respond to conventional treatment. PMID- 9872214 TI - A mediastinal mass in a patient with AIDS. PMID- 9872215 TI - Postpneumonectomy syndrome: recognition and management. AB - STUDY OBJECTIVE: Postpneumonectomy syndrome (PPS) results from extreme shift and rotation of the mediastinum after pneumonectomy producing symptomatic proximal airway obstruction and air trapping. Herein, we review our experience in the treatment of PPS. PATIENTS: Five patients with PPS were treated at our institution between 1991 and 1997. Four patients had previous right pneumonectomy; one patient had left pneumonectomy. Dyspnea was the presenting symptom in all five patients. The time interval to onset of symptoms and to surgical correction ranged from 6 months to 9 years (median: 6 months) and 9 months to 29 years (median, 21 months) after pneumonectomy, respectively. INTERVENTION: The clinical diagnosis of PPS was confirmed with chest radiograph, two-dimensional echocardiography, pulmonary function tests, CT scan, and awake fiberoptic bronchoscopy. Correction of PPS required reexploration of the pneumonectomy space followed by anterior pericardiorrhaphy and insertion of a saline solution-filled Silastic prosthesis (Dow Corning; Midland, MI) for the purpose of correcting the overshift of the mediastinum. There was no morbidity or mortality. RESULTS: All patients had relief of dyspnea. Corrective repositioning of the mediastinum was confirmed by chest radiograph, CT scan, and awake fiberoptic bronchoscopy. There was a mean increase in the cross-sectional diameter, as measured by CT scan, of the obstructed bronchus by 166.7% (range, 100 to 300%) in four patients. One patient had no change in the measured diameter. Postoperatively, the peak expiratory flow rate increased by a mean of 44.2% (range, 40 to 49%) in all five patients. CONCLUSION: The presence of PPS should be considered in all patients presenting with progressive dyspnea after pneumonectomy. Repositioning of the mediastinum with a saline solution-filled prosthesis and anterior pericardiorrhaphy is easily performed and provides immediate and lasting symptomatic relief. PMID- 9872216 TI - Herbicide (Roundup) pneumonitis. AB - A case of acute intoxication presented as toxic pneumonitis after exposure to Roundup (glyphosate) (Solaris Group, Monsanto; San Ramon, CA) herbicide in an agriculture worker. The correct etiologic factor causing this specific clinical picture was identified only 2 weeks later, after a thorough occupational history was taken and meticulous delineation of the working conditions and exposures of the involved worker were made. As a rule, occupational related diseases are not readily elucidated by nonoccupational physicians. However, most acute intoxication events are first encountered by such physicians. In these situations, rapid and comprehensive evaluation is necessary in order to clearly identify the causative agent(s) and to initiate the appropriate treatment. Consulting occupational physicians at this early stage may facilitate early and accurate diagnosis. PMID- 9872217 TI - Percutaneous tracheostomy: is it really better? PMID- 9872218 TI - Lung carcinoma metastasis at the site of central venous access. AB - Tumor metastases rarely are reported in association with central venous catheters. The case reported herein is that of a solid tumor metastasis at the site of a previous indwelling line ipsilateral to a pleural effusion in a patient with non-small cell lung cancer. A review of the literature reveals a single case of intrathoracic malignancy seeding at the site of a central venous catheter. Other investigators are urged to collect information about the development of tumor implants at the site of catheter insertion in patients with cancer and pleural effusions to further define the extent of the problem. PMID- 9872219 TI - Homograft repair of a tuberculous pseudoaneurysm of the ascending aorta. AB - A 34-year-old HIV-positive black man with pulmonary tuberculosis developed progressive widening of the mediastinum. Evaluation by CT and two-dimensional echocardiography demonstrated a large saccular aneurysm of the ascending aorta. This was repaired with a 26-mm aortic homograft using deep hypothermic circulatory arrest. The microbiology of the aneurysm contents showed this to be a tuberculous pseudoaneurysm. PMID- 9872220 TI - Pacing induced mitral regurgitation following radiofrequency ablation of the atrioventricular conduction system: case report and potential mechanism. AB - We describe a patient with hemodynamic deterioration and worsening mitral regurgitation related to right ventricular apex pacing. Time-dependent changes in papillary muscle contraction as well as ventricular remodeling by right ventricular apex pacing might be responsible for this rare but serious complication. PMID- 9872222 TI - Bronchopleural fistula resulting from the use of a thoracic vent: a case report and review. AB - Pneumothorax is defined as the presence of gas or air within the pleural space. Standard treatment is usually based on the evacuation of the gas by various methods. The thoracic vent is a relatively new device used in the treatment of pneumothorax. This report focuses on the first major complication, as far as is known, associated with the use of a thoracic vent. PMID- 9872221 TI - Successful treatment of gemcitabine toxicity with a brief course of oral corticosteroid therapy. AB - Gemcitabine is a nucleoside analog that is useful in the treatment of solid tumors. Its use has been postulated to produce lung injury by causing a capillary leak syndrome. We describe a gemcitabine-treated female patient who developed severe dyspnea, diffuse pulmonary infiltrates, and hypoxia, with evidence of interstitial disease on pulmonary function tests. Following the administration of oral corticosteroids, she had complete resolution of all signs and symptoms of gemcitabine toxicity. Physicians should be aware of this treatable complication of gemcitabine therapy. PMID- 9872223 TI - Congenital bronchoesophageal fistula in an adult. AB - We present a rare case of a congenital bronchoesophageal fistula in a 54-year-old woman with a history of poor feeding tolerance since infancy and repeated pulmonary infections. She initially presented with epigastric and right upper quadrant abdominal pain. Her workup included a barium esophagogram that revealed a fistula between her midesophagus and a left lower lobe segmental bronchus. The fistula was divided, a left lower lobe superior segmentectomy was performed, and an intercostal muscle was placed over the esophageal closure. The patient noted an immediate decrease of postprandial coughing. Congenital respiratory esophageal fistulas that are not associated with esophageal atresia may persist into adulthood before they become clinically apparent. The diagnosis should be considered in certain individuals with suggestive symptomatology and unexplained respiratory pathology. PMID- 9872224 TI - Duropleural fistula manifested as a large pleural transudate: an unusual complication of transthoracic diskectomy. AB - A 52-year-old patient presented with large symptomatic transudative pleural effusion 4 months following T8 to T9 transthoracic diskectomy. Anterior approach diskectomy has replaced the traditional laminectomy for treating symptomatic centrally herniated and calcified thoracic disks. We describe in this report the first case of a large cerebrospinal fluid collection in the pleural cavity caused by persistent postoperative duropleural fistula. The possibility of a duropleural fistula should be entertained in patients who present with a pleural effusion following transthoracic diskectomy. PMID- 9872225 TI - Prevalence of patent foramen ovale and its contribution to hypoxemia in patients with obstructive sleep apnea. PMID- 9872226 TI - A critical commentary and treatise on myocardial repolarization. PMID- 9872227 TI - Recommend caution in defining risk factors for barotrauma in divers. PMID- 9872228 TI - Tracheal lacerations after intubation. PMID- 9872229 TI - Mediastinoscopic treatment of mediastinal cysts. PMID- 9872230 TI - Nasal mask ventilation in children. PMID- 9872231 TI - Comparison of oxygenation in patients staying in Dead Sea or in Jerusalem. PMID- 9872232 TI - Chopin's illness revisited. PMID- 9872233 TI - Methacholine PC20 extrapolation. PMID- 9872234 TI - The third dimension. PMID- 9872235 TI - Cytologically proved malignant pleural effusions. PMID- 9872236 TI - Positivity of pleural fluid cytologic examination in transudative pleural effusions. PMID- 9872237 TI - Real implications of "millennium bug". PMID- 9872238 TI - Achondroplasia: a case of neglect? PMID- 9872239 TI - Early identification of deaf babies. PMID- 9872240 TI - Infertility and pregnancy in epileptic women. PMID- 9872242 TI - Failure of penicillin in Streptococcus pyogenes pharyngeal infection. PMID- 9872241 TI - Rational approach to iron-deficiency anaemia in premenopausal women. PMID- 9872243 TI - Pulmonary valvotomy--50 years ago. PMID- 9872244 TI - Controlled trial of universal neonatal screening for early identification of permanent childhood hearing impairment. Wessex Universal Neonatal Hearing Screening Trial Group. AB - BACKGROUND: Congenital permanent childhood hearing impairment (PCHI) impairs communication skills and, possibly, mental health and employment prospects. Management within 1 year of birth can alleviate most of its adverse effects. Neonatal screening for this disorder is feasible but its benefit for all babies is disputed. We investigated whether neonatal screening of all babies born in hospital, in addition to the standard health visitor distraction test, would increase the rates of early referral, confirmation, and management. METHODS: Between 1993 and 1996, two teams of four part-time testers and equipment moved between two pairs of hospitals to achieve four periods with neonatal screening and four without neonatal screening, each of 4-6 months' duration. Babies did or did not undergo neonatal screening dependent on during which periods they were born. We used a transient evoked otoacoustic emissions test and, in babies who failed this test, an automated auditory brainstem response test on the same day. We referred babies with positive results for audiological assessment. FINDINGS: 53,781 babies were included in the trial, and 25,609 were born during periods with neonatal screening. Neonatal screening achieved 87% coverage of births, with a false-alarm rate of 1.5%, and an overall yield of 90 cases of bilateral PCHI of 40 dB or more relative to hearing threshold level per 100,000 target population, equivalent to 80% of the expected prevalence of the disorder in the population. 71 more babies with moderate or severe PCHI per 100,000 target population were referred before age 6 months during periods with neonatal screening than during periods without. Early confirmation and management of PCHI were significantly increased. The rate of false-negative results from neonatal screening was significantly lower than that for the distraction test (4 vs 27% p=0.041). INTERPRETATION: Neonatal screening is effective in identification of congenital PCHI early and may be particularly useful for babies with moderate and severe PCHI for whom early management may have the most benefit. PMID- 9872245 TI - Cardiovascular and cancer morbidity and mortality and sudden cardiac death in postmenopausal women on oestrogen replacement therapy (ERT) AB - BACKGROUND: Advantages and disadvantages of postmenopausal oestrogen replacement therapy (ERT) are still not clear. We aimed to analyse the relation between postmenopausal oestrogen replacement therapy (ERT), cardiovascular disease, and cancer. METHODS: We examined 7944 women born between 1923 and 1930, who participated in a mammography screening for breast cancer, and who were followed up from 1987 to 1995. The follow-up consisted of 53,305 person-years. 988 women were current users and 757 were former users of ERT. Information about hormone use and health events was obtained through biennial questionnaires and recording and linking information from the hospital discharge registers of the region, the national cancer register, the social insurance reimbursement register, and the national death register. We used proportional-hazards models to calculate risk ratios and 95% CIs, adjusted for eight confounding variables. FINDINGS: Current ERT was associated with decreased cardiovascular mortality and a decrease in sudden cardiac death. Adjusted risk ratio (RR) for cardiovascular mortality in current users was 0.21 (95% CI 0.08-0.59) and in former users 0.75 (0.41-1.37). Absolute risk per 1000 person-years for deaths from acute myocardial infarction (AMI) was 1.1 in never users, 1.2 in former users, and 0.45 in current users (p=0.197). Corresponding absolute risk for other coronary-artery-disease (CAD) deaths was 1.0, 0.81, and 0 (p=0.009), and for deaths from stroke 1.2, 1.0, and 0.15 (p=0.012). Absolute risk for sudden cardiac death was 1.6 in never users, 1.0 in former users, and 0 in current users (p<0.001). Cardiovascular morbidity was not decreased by ERT: the RR for current use was 1.07 (0.86-1.32) and for former use 1.11 (0.89-1.39). Incidence of cardiovascular disease per 1000 person years was 24.9 in never users, 23.4 in former users, and 20.9 in current users (p=0.153). Breast-cancer morbidity did not increase with current ERT--the RR was 0.57 (0.27-1.20). Incidence of breast cancer was 1.8, 1.6, and 1.0 in never, former, and current users (p=0.242). Endometrial cancer increased with current ERT--the RR was 5.06 (2.47-10.41). Incidence of endometrial cancer was 0.52 in never users, 0.51 in former users, and 2.1 in current users (p<0.001). INTERPRETATION: Current ERT reduced primarily sudden cardiac death and predicted reduced cardiovascular mortality, but did not reduce morbidity. ERT did not increase the risk of breast cancer, but was associated with increased risk of endometrial cancer. PMID- 9872246 TI - Age-specific incidence and prevalence rates of treated epilepsy in an unselected population of 2,052,922 and age-specific fertility rates of women with epilepsy. AB - BACKGROUND: There are no data on prevalence or incidence of treated epilepsy, and no data on fertility of women with epilepsy from an unselected UK population. METHODS: We used the General Practice Research Database to ascertain the incidence and prevalence of people with treated epilepsy in an unselected population of 2,052,922 people in England and Wales, and also age-specific fertility rates. We defined period prevalence of treated epilepsy as the number of people with epilepsy taking an antiepileptic drug per 100,000 people during 1995. The incidence of treated epilepsy was defined as the number of new cases of treated epilepsy per 100,000 people during the same period. We calculated fertility rates among women with treated epilepsy between 1991 and 1995 and compared these rates with the population rates for England and Wales in 1993. FINDINGS: The period prevalence of treated epilepsy in 1995 was 5.15 per 1000 people (95% CI 5.05-5.25). The prevalence was lower in children (age 5-9 years 3.16 [2.86-3.48]; 10-14 years 4.05 [3.70-4.42]), and higher in older people (65 69 years 6.01 [5.50-6.57]; 70-74 years 6.53 [5.97-7.14]; 75-79 years 7.39 [6.73 8.11]); 80-84 years 7.54 [6.78-8.39]; 85 years and older 7.73 [6.98-8.66]). The incidence of treated epilepsy was 80.8 per 100,000 people (76.9-84.7). The incidence was lower in children (5-9 years 63.2 [50.5-79.1]; 10-14 years 53.8 [42.4-68.3]) and higher in older people (65-69 years 85.9 [68.5-107.3]; 70-74 years 82.8 [65.0-105.2]; 75-79 years 114.5 [116.9-179.2]; 80-84 years 159 [125.2 202.6]; > or = 85 years 135.4 [100.4-178.7]). Fertility was lower among women with treated epilepsy, with an overall rate of 47.1 livebirths per 1000 women aged 15-44 per year (42.3-52.2), compared with a national rate of 62.6 in the same age-group. The standardised fertility ratios were significantly lower between the ages of 25 and 39 years in women with epilepsy (p<0.001). INTERPRETATION: Compared with previous studies, we found that the incidence of epilepsy was higher in elderly people and lower in children. The prevalence rates also increase with age. Women aged 25-39 years with treated epilepsy have significantly lower fertility rates than those in the general population. Research is needed to identify any potentially preventable causes for the low fertility rates. PMID- 9872247 TI - Prevalence of internalisation-associated gene, prtF1, among persisting group-A streptococcus strains isolated from asymptomatic carriers. AB - BACKGROUND: The failure of antibiotic treatment to eradicate group-A streptococci in up to 30% of patients with pharyngotonsillitis is unexplained. Some strains of group-A streptococci can enter respiratory epithelial cells, where they would be inaccessible to antibiotics unable to penetrate the cell membrane, such as penicillins. The fibronectin-binding proteins, F1 and SfbI, are needed for this process. We hypothesised, therefore, that an intracellular reservoir of group-A streptococci could account, at least partly, for failure to eradicate throat carriage, and that the presence of the gene for fibronectin-binding protein (F1) might be linked to the ability of a strain to persist in the throat after therapy. METHODS: We investigated the frequency of prtF1-containing strains among 67 patients with pharyngotonsillitis. All patients were clinically cured, although 13 of them continued to carry group-A streptococci in the throat during or after therapy. To distinguish between persisting and recolonising strains, isolates from the 13 patients were serologically tested and compared by polymorphic DNA-amplification technique. FINDINGS: 12 (92%) of the 13 patients with symptomless carriage had prtF1-containing strains in the throat, compared with 16 (30%) of the 54 patients with successful eradication (p=0.0001). Three of the 13 eradication-failure patients were recolonised with strains that differed from the pretreatment strains. Nine of the ten (90%) persisting strains carried prtF1 (p=0.0009). INTERPRETATION: Our findings suggest that protein-F1-mediated entry to cells is involved in the causative process of the carriage state. PMID- 9872249 TI - A 70-year-old man with an empyema. PMID- 9872248 TI - Effect of angiotensin-converting-enzyme (ACE) inhibitor trandolapril on human diabetic neuropathy: randomised double-blind controlled trial. AB - BACKGROUND: Diabetes is a common cause of polyneuropathy. The development and progression of nephropathy, retinopathy, and neuropathy are closely related. Angiotensin-converting enzyme (ACE) inhibitors delay progression of both nephropathy and retinopathy. We investigated the effect of ACE inhibition on diabetic neuropathy. METHODS: We recruited 41 normotensive patients with type I or type II diabetes and mild neuropathy into a randomised double-blind placebo controlled trial. Changes in the neuropathy symptom and deficit scores, vibration perception threshold, peripheral-nerve electrophysiology, and cardiovascular autonomic function, were assessed at 6 and 12 months. The primary endpoint was the change in peroneal nerve motor conduction velocity. FINDINGS: We found no significant difference at baseline for age, HbA1c, blood pressure, or severity of neuropathy between two groups. There was no change in HbA1c over the treatment period. Peroneal motor nerve conduction velocity (p=0.03) and M-wave amplitude (p=0.03) increased, and the F-wave latency (p=0.03) decreased and sural nerve action potential amplitude increased (p=0.04) significantly after 12 months of treatment with trandolapril compared with placebo. Vibration-perception threshold, autonomic function, and the neuropathy symptom and deficit score showed no improvement in either group. INTERPRETATION: The ACE inhibitor trandolapril may improve peripheral neuropathy in normotensive patients with diabetes. Larger clinical trials are needed to confirm these data before changes to clinical practice can be advocated. PMID- 9872250 TI - Immune constitution of complete DiGeorge anomaly by transplantation of unmobilised blood mononuclear cells. PMID- 9872251 TI - Long-term outcome after thalamotomy for movement disorders in multiple sclerosis. PMID- 9872252 TI - Gastroparesis as a cause of nausea and vomiting after high-dose chemotherapy and haemopoietic stem-cell transplantation. PMID- 9872253 TI - Neonatal varicella despite maternal immunity. PMID- 9872254 TI - Case-control study of interactions between genetic and environmental factors in Parkinson's disease. PMID- 9872255 TI - Antioxidant effects of herbs in Crete. PMID- 9872256 TI - Changing belief in obstetrics: impact of two multicentre randomised controlled trials. PMID- 9872258 TI - Secular and seasonal variation of length and weight at birth. PMID- 9872257 TI - High incidence of Creutzfeldt-Jakob disease in rural Calabria, Italy. PMID- 9872259 TI - UK "white paper on tobacco" welcomed. PMID- 9872260 TI - Festive drinking's slippery slope beckons. PMID- 9872261 TI - Mental care versus public safety in the UK. PMID- 9872262 TI - Wakley prize essay. Good times. PMID- 9872264 TI - Art and neuroscience: how the brain sees Vermeer's Woman Holding a Balance. PMID- 9872263 TI - 175th anniversary lecture. Medical journals and the shaping of medical knowledge. PMID- 9872265 TI - Jealousy and mutilation: nose-biting as retribution for adultery. PMID- 9872266 TI - Apgar score and Soranus of Ephesus. PMID- 9872267 TI - Why leeches influence my physical examination. PMID- 9872268 TI - Helicobacter pylori eradication in long-term users of non-steroidal anti inflammatory drugs. PMID- 9872269 TI - Helicobacter pylori eradication in long-term users of non-steroidal anti inflammatory drugs. PMID- 9872270 TI - Helicobacter pylori eradication in long-term users of non-steroidal anti inflammatory drugs. PMID- 9872271 TI - Paramedic direct admission of heart-attack patients to coronary care unit. PMID- 9872272 TI - Paramedic direct admission of heart-attack patients to coronary care unit. PMID- 9872273 TI - Community-acquired pneumonia. PMID- 9872274 TI - Community-acquired pneumonia. PMID- 9872275 TI - Association of road-traffic accidents with benzodiazepine use. PMID- 9872276 TI - Simvastatin and thrombotic thrombocytopenic purpura. PMID- 9872277 TI - REIN follow-up trial. Ramipril Efficacy in Nephropathy. PMID- 9872278 TI - REIN follow-up trial. Ramipril Efficacy in Nephropathy. PMID- 9872279 TI - Mycoplasma hominis parasitism of Trichomonas vaginalis. PMID- 9872280 TI - Autoregulation of cerebral blood flow. PMID- 9872281 TI - Nicaragua's debt burden: a permanent hurricane. PMID- 9872282 TI - Iodine-rich drinking water of natural origin in China. PMID- 9872283 TI - Culling of badgers and control of bovine tuberculosis. PMID- 9872284 TI - Jonathan Mann's mantle. PMID- 9872285 TI - Why change drug names? PMID- 9872286 TI - Record keeping and compassionate care. PMID- 9872287 TI - Rationing of health care. PMID- 9872288 TI - Leadership at Mbarara. PMID- 9872289 TI - Below the belt. PMID- 9872290 TI - Medical iconography in the 17th century. PMID- 9872291 TI - The Nobel chronicles. 1931: Otto Heinrich Warburg (1883-1970). PMID- 9872293 TI - Protest in Paris. PMID- 9872292 TI - Setting a bad example on AIDS. PMID- 9872294 TI - More foreign research students stay in US to cash in on boom. PMID- 9872295 TI - UN stands up for East German scientists. PMID- 9872296 TI - Search hopes to detect laser signals from extraterrestrial life. PMID- 9872297 TI - German standstill as fewer chairs available. PMID- 9872298 TI - Israeli industry asks for boost to R&D. PMID- 9872299 TI - Protest at Nobel omission of Moncada. PMID- 9872300 TI - Protest at Nobel omission of Moncada. PMID- 9872301 TI - Protest at Nobel omission of Moncada. PMID- 9872302 TI - Bitter pill to swallow over medical education. PMID- 9872303 TI - Benchmarking international research. PMID- 9872304 TI - Genome sequencing. The worm revealed. PMID- 9872305 TI - Structural biology. The ABC of a versatile engine. PMID- 9872307 TI - Apoptosis. Death deceiver. PMID- 9872306 TI - Antigen presentation. A protease draws first blood. PMID- 9872308 TI - The thymus in the age of retirement. PMID- 9872309 TI - Trick or treat from food endocannabinoids? PMID- 9872310 TI - Immobile plasticizer in flexible PVC. PMID- 9872311 TI - Genetic instabilities in human cancers. AB - Whether and how human tumours are genetically unstable has been debated for decades. There is now evidence that most cancers may indeed be genetically unstable, but that the instability exists at two distinct levels. In a small subset of tumours, the instability is observed at the nucleotide level and results in base substitutions or deletions or insertions of a few nucleotides. In most other cancers, the instability is observed at the chromosome level, resulting in losses and gains of whole chromosomes or large portions thereof. Recognition and comparison of these instabilities are leading to new insights into tumour pathogenesis. PMID- 9872312 TI - Coexisting conical bipolar and equatorial outflows from a high-mass protostar. AB - The BN/KL region in the Orion molecular cloud is an archetype for the study of the formation of stars much more massive than the Sun. This region contains luminous young stars and protostars but, like most star-forming regions, is difficult to study in detail because of the obscuring effects of dust and gas. Our basic expectations are shaped to some extent by the present theoretical picture of star formation, the cornerstone of which is that protostars accrete gas from rotating equatorial disks and shed angular momentum by ejecting gas in bipolar outflows. The main source of the outflow in the BN/KL region may be an object known as radio source I, which is commonly believed to be surrounded by a rotating disk of molecular material. Here we report high-resolution observations of silicon monoxide (SiO) and water maser emission from the gas surrounding source I. We show that within 60 AU of the source (about the size of the Solar System), the region is dominated by a conical bipolar outflow, rather than the expected disk. A slower outflow, close to the equatorial plane of the protostellar system, extends to radii of 1,000 AU. PMID- 9872313 TI - Polysaccharide elasticity governed by chair-boat transitions of the glucopyranose ring. AB - Many common, biologically important polysaccharides contain pyranose rings made of five carbon atoms and one oxygen atom. They occur in a variety of cellular structures, where they are often subjected to considerable tensile stress. The polysaccharides are thought to respond to this stress by elastic deformation, but the underlying molecular rearrangements allowing such a response remain poorly understood. It is typically assumed, however, that the pyranose ring structure is inelastic and locked into a chair-like conformation. Here we describe single molecule force measurements on individual polysaccharides that identify the pyranose rings as the structural unit controlling the molecule's elasticity. In particular, we find that the enthalpic component of the polymer elasticity of amylose, dextran and pullulan is eliminated once their pyranose rings are cleaved. We interpret these observations as indicating that the elasticity of the three polysaccharides results from a force-induced elongation of the ring structure and a final transition from a chair-like to a boat-like conformation. We expect that the force-induced deformation of pyranose rings reported here plays an important role in accommodating mechanical stresses and modulating ligand binding in biological systems. PMID- 9872314 TI - Mice lacking melanin-concentrating hormone are hypophagic and lean. AB - Feeding is influenced by hypothalamic neuropeptides that promote (orexigenic peptides) or inhibit feeding. Of these, neuropeptide Y (NPY) in the arcuate nucleus and melanin-concentrating hormone (MCH) and orexins/hypocretins in the lateral hypothalamus have received attention because their expression is increased during fasting and because they promote feeding when administered centrally. Surprisingly, absence of the orexigenic neuropeptide NPY fails to alter feeding or body weight in normal mice. As deficiency of a single component of the pathway that limits food intake (such as leptin or receptors for melanocortin-4) causes obesity, it has been suggested that orexigenic signals are more redundant than those limiting food intake. To define further the physiological role of MCH and to test the redundancy of orexigenic signals, we generated mice carrying a targeted deletion of the MCH gene. MCH-deficient mice have reduced body weight and leanness due to hypophagia (reduced feeding) and an inappropriately increased metabolic rate, despite their reduced amounts of both leptin and arcuate nucleus pro-opiomelanocortin messenger RNA. Our results show that MCH is a critical regulator of feeding and energy balance which acts downstream of leptin and the melanocortin system, and that deletion of a gene encoding a single orexigenic peptide can result in leanness. PMID- 9872315 TI - GABA(B) receptors function as a heteromeric assembly of the subunits GABA(B)R1 and GABA(B)R2. AB - The principal inhibitory neurotransmitter GABA (gamma-aminobutyric acid) exerts its effects through two ligand-gated channels, GABA(A) and GABA(C) receptors, and a third receptor, GABA(B) , which acts through G proteins to regulate potassium and calcium channels. Cells heterologously expressing the cloned DNA encoding the GABA(B)R1 protein exhibit high-affinity antagonist-binding sites, but they produce little of the functional activity expected from studies of endogenous GABA(B) receptors in the brain. Here we describe a new member of the GABA(B) polypeptide family, GABA(B)R2, that shows sequence homology to GABA(B)R1. Neither GABA(B)R1 nor GABA(B)R2, when expressed individually, activates GIRK-type potassium channels; however, the combination of GABA(B)R1 and GABA(B)R2 confers robust stimulation of channel activity. Both genes are co-expressed in individual neurons, and both proteins co-localize in transfected cells. Moreover, immunoprecipitation experiments indicate that the two polypeptides associate with each other, probably as heterodimers. Several G-protein-coupled receptors (GPCRs) exist as high-molecular-weight species, consistent with the formation of dimers by these receptors, but the relevance of these species for the functioning of GPCRs has not been established. We have now shown that co-expression of two GPCR structures, GABA(B)R1 and GABA(B)R2, belonging to the same subfamily is essential for signal transduction by GABA(B) receptors. PMID- 9872316 TI - Heterodimerization is required for the formation of a functional GABA(B) receptor. AB - GABA (gamma-aminobutyric acid) is the main inhibitory neurotransmitter in the mammalian central nervous system, where it exerts its effects through ionotropic (GABA(A/C)) receptors to produce fast synaptic inhibition and metabotropic (GABA(B)) receptors to produce slow, prolonged inhibitory signals. The gene encoding a GABA(B) receptor (GABA(B)R1) has been cloned; however, when expressed in mammalian cells this receptor is retained as an immature glycoprotein on intracellular membranes and exhibits low affinity for agonists compared with the endogenous receptor on brain membranes. Here we report the cloning of a complementary DNA encoding a new subtype of the GABAB receptor (GABA(B)R2), which we identified by mining expressed-sequence-tag databases. Yeast two-hybrid screening showed that this new GABA(B)R2-receptor subtype forms heterodimers with GABA(B)R1 through an interaction at their intracellular carboxy-terminal tails. Upon expression with GABA(B)R2 in HEK293T cells, GABA(B)R1 is terminally glycosylated and expressed at the cell surface. Co-expression of the two receptors produces a fully functional GABA(B) receptor at the cell surface; this receptor binds GABA with a high affinity equivalent to that of the endogenous brain receptor. These results indicate that, in vivo, functional brain GABA(B) receptors may be heterodimers composed of GABA(B)R1 and GABA(B)R2. PMID- 9872317 TI - GABA(B)-receptor subtypes assemble into functional heteromeric complexes. AB - B-type receptors for the neurotransmitter GABA (gamma-aminobutyric acid) inhibit neuronal activity through G-protein-coupled second-messenger systems, which regulate the release of neurotransmitters and the activity of ion channels and adenylyl cyclase. Physiological and biochemical studies show that there are differences in drug efficiencies at different GABA(B) receptors, so it is expected that GABA(B)-receptor (GABA(B)R) subtypes exist. Two GABA(B)-receptor splice variants have been cloned (GABA(B)R1a and GABA(B)R1b), but native GABA(B) receptors and recombinant receptors showed unexplained differences in agonist binding potencies. Moreover, the activation of presumed effector ion channels in heterologous cells expressing the recombinant receptors proved difficult. Here we describe a new GABA(B) receptor subtype, GABA(B)R2, which does not bind available GABA(B) antagonists with measurable potency. GABA(B)R1a, GABA(B)R1b and GABA(B)R2 alone do not activate Kir3-type potassium channels efficiently, but co-expression of these receptors yields a robust coupling to activation of Kir3 channels. We provide evidence for the assembly of heteromeric GABA(B) receptors in vivo and show that GABA(B)R2 and GABA(B)R1a/b proteins immunoprecipitate and localize together at dendritic spines. The heteromeric receptor complexes exhibit a significant increase in agonist- and partial-agonist-binding potencies as compared with individual receptors and probably represent the predominant native GABA(B) receptor. Heteromeric assembly among G-protein-coupled receptors has not, to our knowledge, been described before. PMID- 9872318 TI - Moderate loss of function of cyclic-AMP-modulated KCNQ2/KCNQ3 K+ channels causes epilepsy. AB - Epilepsy affects more than 0.5% of the world's population and has a large genetic component. It is due to an electrical hyperexcitability in the central nervous system. Potassium channels are important regulators of electrical signalling, and benign familial neonatal convulsions (BFNC), an autosomal dominant epilepsy of infancy, is caused by mutations in the KCNQ2 or the KCNQ3 potassium channel genes. Here we show that KCNQ2 and KCNQ3 are distributed broadly in brain with expression patterns that largely overlap. Expression in Xenopus oocytes indicates the formation of heteromeric KCNQ2/KCNQ3 potassium channels with currents that are at least tenfold larger than those of the respective homomeric channels. KCNQ2/KCNQ3 currents can be increased by intracellular cyclic AMP, an effect that depends on an intact phosphorylation site in the KCNQ2 amino terminus. KCNQ2 and KCNQ3 mutations identified in BFNC pedigrees compromised the function of the respective subunits, but exerted no dominant-negative effect on KCNQ2/KCNQ3 heteromeric channels. We predict that a 25% loss of heteromeric KCNQ2/KCNQ3 channel function is sufficient to cause the electrical hyperexcitability in BFNC. Drugs raising intracellular cAMP may prove beneficial in this form of epilepsy. PMID- 9872319 TI - Changes in thymic function with age and during the treatment of HIV infection. AB - The thymus represents the major site of the production and generation of T cells expressing alphabeta-type T-cell antigen receptors. Age-related involution may affect the ability of the thymus to reconstitute T cells expressing CD4 cell surface antigens that are lost during HIV infection; this effect has been seen after chemotherapy and bone-marrow transplantation. Adult HIV-infected patients treated with highly active antiretroviral therapy (HAART) show a progressive increase in their number of naive CD4-positive T cells. These cells could arise through expansion of existing naive T cells in the periphery or through thymic production of new naive T cells. Here we quantify thymic output by measuring the excisional DNA products of TCR-gene rearrangement. We find that, although thymic function declines with age, substantial output is maintained into late adulthood. HIV infection leads to a decrease in thymic function that can be measured in the peripheral blood and lymphoid tissues. In adults treated with HAART, there is a rapid and sustained increase in thymic output in most subjects. These results indicate that the adult thymus can contribute to immune reconstitution following HAART. PMID- 9872320 TI - An asparaginyl endopeptidase processes a microbial antigen for class II MHC presentation. AB - Foreign protein antigens must be broken down within endosomes or lysosomes to generate suitable peptides that will form complexes with class II major histocompatibility complex molecules for presentation to T cells. However, it is not known which proteases are required for antigen processing. To investigate this, we exposed a domain of the microbial tetanus toxin antigen (TTCF) to disrupted lysosomes that had been purified from a human B-cell line. Here we show that the dominant processing activity is not one of the known lysosomal cathepsins, which are generally believed to be the principal enzymes involved in antigen processing, but is instead an asparagine-specific cysteine endopeptidase. This enzyme seems similar or identical to a mammalian homologue of the legumain/haemoglobinase asparaginyl endopeptidases found originally in plants and parasites. We designed competitive peptide inhibitors of B-cell asparaginyl endopeptidase (AEP) that specifically block its proteolytic activity and inhibit processing of TTCF in vitro. In vivo, these inhibitors slow TTCF presentation to T cells, whereas preprocessing of TTCF with AEP accelerates its presentation, indicating that this enzyme performs a key step in TTCF processing. We also show that N-glycosylation of asparagine residues blocks AEP action in vitro. This indicates that N-glycosylation could eliminate sites of processing by AEP in mammalian proteins, allowing preferential processing of microbial antigens. PMID- 9872321 TI - Genomic amplification of a decoy receptor for Fas ligand in lung and colon cancer. AB - Fas ligand (FasL) is produced by activated T cells and natural killer cells and it induces apoptosis (programmed cell death) in target cells through the death receptor Fas/Apol/CD95. One important role of FasL and Fas is to mediate immune cytotoxic killing of cells that are potentially harmful to the organism, such as virus-infected or tumour cells. Here we report the discovery of a soluble decoy receptor, termed decoy receptor 3 (DcR3), that binds to FasL and inhibits FasL induced apoptosis. The DcR3 gene was amplified in about half of 35 primary lung and colon tumours studied, and DcR3 messenger RNA was expressed in malignant tissue. Thus, certain tumours may escape FasL-dependent immune-cytotoxic attack by expressing a decoy receptor that blocks FasL. PMID- 9872322 TI - Crystal structure of the ATP-binding subunit of an ABC transporter. AB - ABC transporters (also known as traffic ATPases) form a large family of proteins responsible for the translocation of a variety of compounds across membranes of both prokaryotes and eukaryotes. The recently completed Escherichia coli genome sequence revealed that the largest family of paralogous E. coli proteins is composed of ABC transporters. Many eukaryotic proteins of medical significance belong to this family, such as the cystic fibrosis transmembrane conductance regulator (CFTR), the P-glycoprotein (or multidrug-resistance protein) and the heterodimeric transporter associated with antigen processing (Tap1-Tap2). Here we report the crystal structure at 1.5 A resolution of HisP, the ATP-binding subunit of the histidine permease, which is an ABC transporter from Salmonella typhimurium. We correlate the details of this structure with the biochemical, genetic and biophysical properties of the wild-type and several mutant HisP proteins. The structure provides a basis for understanding properties of ABC transporters and of defective CFTR proteins. PMID- 9872323 TI - Gads is a novel SH2 and SH3 domain-containing adaptor protein that binds to tyrosine-phosphorylated Shc. AB - Shc proteins are important substrates of receptor and cytoplasmic tyrosine kinases that couple activated receptors to downstream signaling enzymes. Phosphorylation of Shc tyrosine residues 239 and 317 leads to recruitment of the Grb2-Sos complex, thus linking Shc phosphorylation to Ras activation. We have used phosphorylated peptides corresponding to the regions spanning tyrosine 239/240 and 317 of Shc in an expression library screen to identify additional downstream targets of Shc. Here we report the identification of Gads, a novel adaptor protein most similar to Grb2 and Grap that contains amino and carboxy terminal SH3 domains flanking a central SH2 domain and a 120 amino acid unique region. Gads is most highly expressed in the thymus and spleen of adult animals and in human leukemic cell lines. The binding specificity of the Gads SH2 domain is similar to Grb2 and mediates the interaction of Gads with Shc, Bcr-Abl and c kit. Gads does not interact with Sos, Cbl or Sam68, although the isolated carboxy terminal Gads SH3 domain is able to bind these molecules in vitro. Our results suggest that the unique structure of Gads regulates its interaction with downstream SH3 domain-binding proteins and that Gads may function to couple tyrosine-phosphorylated proteins such as Shc, Bcr-Abl and activated receptor tyrosine kinases to downstream effectors distinct from Sos and Ras. PMID- 9872324 TI - Primary keratinocytes have an adhesion dependent S phase checkpoint that is absent in immortalized cell lines. AB - In order to understand the mechanism through which loss of anchorage inhibits growth, we have investigated the events that occur in murine keratinocytes upon substratum detachment utilizing both primary cells and established immortalized cell lines. Our data has revealed that while both primary and immortalized cells undergo growth arrest in suspension, the nature of this arrest is markedly different. Primary cells exhibit a growth arrest that is characterized by rapid cessation of DNA synthesis resulting in a static S phase population. In contrast, an immortalized non-tumorgenic cell line, Balb MK, exhibits growth arrest as measured by thymidine incorporation, but does not prevent cells that have entered S phase from continuing into G2/M, and accumulating as a 4N population. In contrast to both primary and MK cells, the tumorigenic SLC-1 cell line did not accumulate in a specific cell cycle interval and were able to undergo continuous growth in suspension. Examination of cyclin A protein and its associated activity revealed that cyclin A protein levels decreased in primary but not MK cells; suggesting the continued presence of cyclin A may allow continued DNA synthesis observed in MK cells. Furthermore, we demonstrate the accumulation of suspension cultured MK cells as a 4N population correlated with the loss of cyclin A/cdk2 kinase activity, which in turn occurred through the accumulation of p27kip1, whereas neither p27kip1 accumulation nor loss of cyclin A activity was observed in SLC-1 cells. Our results clearly reveal that the process of growth inhibition in suspension cultured cells may occur in several forms with distinct characteristics that are dependent on the status of cyclin/cdk complexes and CKI proteins. Tumor derived cells in suspension did not lose cyclin A dependent kinase activity and thus continued to grow and divide. PMID- 9872325 TI - Upregulation of cyclin T1/CDK9 complexes during T cell activation. AB - Cyclin T1 has been identified recently as a regulatory subunit of CDK9 and as a component of the transcription elongation factor P-TEFb. Cyclin T1/CDK9 complexes phosphorylate the carboxy terminal domain (CTD) of RNA polymerase II (RNAP II) in vitro. Here we report that the levels of cyclin T1 are dramatically upregulated by two independent signaling pathways triggered respectively by PMA and PHA in primary human peripheral blood lymphocytes (PBLs). Activation of these two pathways in tandem is sufficient for PBLs to enter and progress through the cell cycle. However, the expression of cyclin T1 is not growth and/or cell cycle regulated in other cell types, indicating that regulation of cyclin T1 expression is dependent on tissue-specific signaling pathways. Upregulation of cyclin T1 in stimulated PBLs results in induction of the CTD kinase activity of the cyclin T1/CDK9 complex, which in turn correlates directly with phosphorylation of RNAP II in vivo, linking for the first time activation of the cyclin T1/ CDK9 pair with phosphorylation of RNAP II in vivo. In addition, we report here that endogenous CDK9 and cyclin T1 complexes associate with HIV-1 generated Tat in relevant cells and under physiological conditions (HIV-1 infected T cells). This, together with our results showing that HIV-1 replication in stimulated PBLs correlates with the levels of cyclin T1 protein and associated CTD kinase activity, suggests that the cyclin T1/CDK9 pair is one of the HIV-1 required host cellular cofactors generated during T cell activation. PMID- 9872326 TI - Transactivation of the human cdc2 promoter by adenovirus E1A in cycling cells is mediated by induction of a 110-kDa CCAAT-box-binding factor. AB - Cyclin-dependent protein kinases (Cdks) are key regulatory proteins of the eukaryotic cell cycle. Cdc2 is expressed in late G1/S phase and functions in the G2 to M phase transition. Adenovirus E1A proteins are known to induce the expression of p34cdc2 and DNA synthesis in normal quiescent cells. In this study, mutational analysis of the human cdc2 promoter revealed that transactivation of the promoter by the E1A proteins in cycling cells is mediated through the two CCAAT box binding motifs. A 110-kDa protein (CBF/cdc2) was identified in nuclear extracts from monkey kidney (CV-1) cells stably expressing E1A as well as from adenovirus-transformed human 293 cells. Further, we show that this EIA-inducible CBF/cdc2 is related to the CBF which was shown to activate the heat shock protein 70 promoter. Analyses of the functional domain(s) of E1A required for the induction of the CBF and transactivation of the cdc2 promoter in these conditions revealed that E1A mutants which were defective in binding the pRB family of proteins or the cellular p300 protein were still active in assays measuring the induction of the CBF and transactivation of the cdc2 promoter, albeit with reduced efficiencies. But the E1A mutant which lost both functional domains was inactive in these assays. These results suggest that E1A has redundant functional domains for the induction of the 110-kDa CBF and activation of human cdc2 gene expression. PMID- 9872327 TI - A targeted disruption of the murine Brca1 gene causes gamma-irradiation hypersensitivity and genetic instability. AB - Germline mutations of the Brcal gene are responsible for most cases of familial breast and ovarian cancers, but somatic mutations are rarely detected in sporadic events. Moreover, mouse embryos deficient for Brca1 have been shown to die during early embryogenesis due to a proliferation defect. These findings seem incompatible with the tumor suppress function assigned to this gene and raise questions about the mechanism by which Brca1 mutations cause tumorigenesis. We now directly demonstrate that BRCA1 is responsible for the integrity of the genome. Murine embryos carrying a Brca1 null mutation are developmentally retarded and hypersensitive to gamma-irradiation, suggesting a failure in DNA damage repair. This notion is supported by spectral karyotyping (SKY) of metaphase chromosomes, which display numerical and structural aberrations. However, massive chromosomal abnormalities are only observed when a p53-/- background is introduced. Thus, a p53 dependent cell cycle checkpoint arrests the mutant embryos and prevents the accumulation of damaged DNA. Brca1-/- fibroblasts are not viable, nor are Brca1-/-:p53-/- fibroblasts. However, proliferative foci arise from Brca1-/-: p53-/- cells, probably due to additional mutations that are a consequence of the accumulating DNA damage. We believe that the increased incidence of such additional mutations accounts for the mechanism of tumorigenesis associated with Brca1 mutations in humans. PMID- 9872328 TI - Molecular and immunological analysis of genetic prostate specific antigen (PSA) vaccine. AB - Nucleic acid immunization has been investigated as immunotherapy for infectious diseases as well as for treating specific types of cancers. In this approach, nucleic acid expression cassettes are directly inoculated into the host, whose transfected cells become the production source of novel and possibly immunologically foreign protein. We have developed a DNA vaccine construct which encodes for PSA by cloning a cDNA for PSA into a mammalian expression vector under control of a CMV promoter. We investigated and characterized the immunogenicity of PSA DNA expression cassettes in mice. PSA-specific immune responses induced in vivo by immunization were characterized by enzyme-linked immunosorbent assay (ELISA), T helper proliferation cytotoxic T lymphocyte (CTL), and flow cytometry assays. We observed a strong and persistent antibody response against PSA for at least 180 days following immunization. In addition, a significant T helper cell proliferation was observed against PSA protein. Using synthetic peptides spanning the PSA open frame, we identified four dominant T helper epitopes of PSA. Furthermore, immunization with PSA plasmid induced MHC Class I CD8+ T cell-restricted cytotoxic T lymphocyte response against tumor cell targets expressing PSA. The prostate represents a very specific functional organ critical for reproduction but not for the health and survival of the individual. Understanding the immunogenicity of PSA DNA immunization cassettes offers insight into the possible use of this tumor-associated antigen as a target for immunotherapy. These results demonstrate the ability of the genetic PSA to serve as a specific immune target capable of generating both humoral and cellular immune responses in vivo. PMID- 9872329 TI - The p16 (CDKN2A) gene is involved in the growth of neuroblastoma cells and its expression is associated with prognosis of neuroblastoma patients. AB - We previously reported that loss of heterozygosity (LOH) on chromosome 9p21 correlates with poor prognosis of neuroblastoma and the p16 gene is not expressed in approximately two thirds of neuroblastoma cell lines. Here we demonstrated that p16 expression was induced by 5-aza-2-deoxycytidine treatment in cell lines with 5' CpG island methylation but not in cell lines without methylation. Furthermore, the cell cycle of neuroblastoma cell lines significantly delayed with accumulation of cells in G1 phase by transfection of a wild-type p16 expression vector. These results indicate that p16 is inactivated in part by DNA methylation and its expression is involved in the growth of neuroblastoma cells in vitro. To assess the biological and clinical significance of p16 expression in primary tumors, we undertook immunohistochemical analysis in 74 paraffin sections of neuroblastomas. p16 protein was undetectable in 45 of 74 cases (61%) and lack of p16 expression significantly correlated with poor prognosis of patients and advanced stage of the disease. There was no correlation between loss of p16 expression and N-myc amplification in these tumors. These results indicate that inactivation of the p16 gene is involved in the progression of neuroblastoma independently of N-myc amplification. PMID- 9872331 TI - Repression of Stat3 activity by activation of mitogen-activated protein kinase (MAPK). AB - STAT proteins are activated by phosphorylation at specific tyrosine residue at the carboxy-terminus which is required for dimer-formation, nuclear translocation, DNA binding and transcriptional activity in cells treated with cytokines and growth factors. Recent studies have indicated that STATs are also phosphorylated by MAPK, or extracellular signal-regulated kinase (ERK) on serine. We investigated the role of ERK on the regulation of STAT activity. Here, we report that ERK2 activated by its upstream kinase, MEK1, represses Stat3 transcriptional activity induced by Src or Jak-2. To unravel the mechanism of repression, we further showed that Stat3 DNA binding activity and its tyrosine phosphorylation are also inhibited under the same conditions. ERK2 phosphorylates Stat3 on three serine-containing peptides and decreases its tyrosine phosphorylation induced by EGF treatment. We also detected an association of ERK2 and Stat3 in vivo which is modulated positively by activation of ERK2, but negatively by Jak2. We propose that MAP kinase cascade may negatively regulate Stat3 activities by decreasing its tyrosine phosphorylation and also possibly by association. PMID- 9872330 TI - Nrf2 and Nrf1 in association with Jun proteins regulate antioxidant response element-mediated expression and coordinated induction of genes encoding detoxifying enzymes. AB - Antioxidant response element (ARE)-mediated expression and coordinated induction of genes encoding detoxifying enzymes is one mechanism of critical importance to cellular protection against oxidative stress. In the present report, we demonstrate that nuclear transcription factors Nrf2 and Nrf1 associate with Jun (c-Jun, Jun-B and Jun-D) proteins to upregulate ARE-mediated expression and coordinated induction of detoxifying enzymes in response to antioxidants and xenobiotics. Nrf-Jun association/heterodimerization and binding to the ARE required unknown cytosolic factor(s). Nrf2 containing one mutated leucine in its leucine zipper region was more efficient in upregulation of ARE-mediated gene expression, as compared to Nrf1 with two mutated leucines. PMID- 9872332 TI - Down-regulation of BRCA1 in human sporadic breast cancer; analysis of DNA methylation patterns of the putative promoter region. AB - Germ-line alterations of BRCA1 are responsible for about 50% of familial breast cancers. Although its biological function(s) has not yet been fully determined, it has been suggested that it may act as a tumor suppressor gene in breast and ovarian cancers. In sporadic breast cancers alterations of BRCA1 have not been detected and in vitro experiments have indicated that BRCA1 negatively regulates cellular proliferation. The present study was designed to identify and quantify, the BRCA1 mRNA levels, in normal and neoplasic human breast tissue. BRCA1 mRNA molecules were quantified using competitive reverse transcriptase PCR assays. DNA methylation patterns of this gene have been analysed by Southern blot experiments using methylation sensitive restriction enzymes. We found that BRCA1 mRNA levels were significantly lower in sporadic breast cancers (37 cases analysed, 24 cases of invasive ductal carcinomas not otherwise specified (NOS), two lobular carcinomas in situ two medullary carcinomas, four invasive lobular carcinomas, two invasive mucinous carcinomas and three invasive ductal carcinomas with predominantly in situ component) compared with normal breast tissues (P=0.0003). This down-regulation of BRCA1 is observed in all histologic types analysed. In invasive ductal carcinomas NOS, this down-regulation does not correlate with any of the prognostic factors studied (tumor size, node status, histologic grade, hormone receptor status). In the samples analysed, alterations of DNA methylation patterns were not dectected in the vicinity of the major transcription start site. These data suggest the involvement of BRCA1 in the carcinogenesis of these histologic types. PMID- 9872333 TI - Dual effect of erbB-2 depletion on the regulation of DNA repair and cell cycle mechanisms in non-small cell lung cancer cells. AB - Overexpression of the erbB-2 tyrosine kinase receptor, p185erbB-2, is a common alteration in non-small cell lung cancer (NSCLC) and has been associated with poor prognosis and a tumor drug resistance phenotype. In this study, we have examined the consequences of erbB-2 depletion on DNA repair, cell cycle, and apoptosis using a panel of NSCLC cell lines constitutively overexpressing erbB-2 receptor. Depletion of the erbB-2 was achieved using the tyrosine kinase inhibitor CP127,374 which promotes erbB-2 degradation. Treatment with CP127,374 concentrations which deplete erbB-2 and inhibit tyrosine phosphorylation resulted in downregulation of DNA repair mechanisms and cell accumulation at G1 phase of the cell cycle. GI arrest was observed in cells with mutated p53 as well as cells lacking p53 protein, suggesting a p53-independent mechanisms. NSCLC cells which overexpress erbB-2 were more resistant to cisplatin-induced cytotoxicity in comparison to cells expressing low levels of erbB-2. Treatment with CP127,374 alone did not result in any induction of apoptosis. A combination of CP127,374 and cisplatin, however, was more potent in cell growth inhibition and induction of apoptosis compared to treatment with cisplatin alone. Together, our results further support a pivotal role of erbB-2 signaling in the regulatory balance between DNA repair, cell cycle checkpoints and apoptosis; all these mechanisms are essential determinants for tumor cell destiny following chemotherapy stress. PMID- 9872334 TI - Isolation of growth suppressors from a cDNA expression library. AB - We describe an experimental procedure for the isolation of growth inhibitory sequences from a complex cDNA library. This approach first takes advantage of the SETGAP technique (selectable expression of transient growth arrest phenotype) to enrich for growth inhibitory sequences, followed by a screening procedure to identify individual cDNAs that inhibit cell proliferation. Here we provide a detailed description of the experimental protocol and report the characterization of two cDNA sequences isolated in our initial screen of a mouse cDNA library. One of these cDNAs encodes the mouse ubiquitin-conjugation enzyme UbcM2. The other encodes a truncated form of a novel WD40 repeat protein, named Bopl, which is conserved from yeast to human. Together, these results demonstrate a new approach for the isolation of growth suppressors from cDNA libraries, and identify a previously unknown gene likely to be involved in growth control. PMID- 9872336 TI - Experimental equations to calculate aminoglycoside clearances through cuprophan dialyzers. AB - Aminoglycosides are a group of antibiotics of particular interest because of their widespread use in the treatment of infections caused by gram-negative bacteria. However, they are difficult to dose accurately because of their narrow therapeutic range and overdosing produces a large number of toxic effects. This paper describes a study carried out with the aim of ensuring the accurate administration of these antibiotics in end-stage renal disease patients undergoing hemodialysis, by developing equations which enable the dosage for individual patients to be calculated according to the dialyzer being used and the operational conditions set up during the dialysis session. PMID- 9872335 TI - The LMO1 and LDB1 proteins interact in human T cell acute leukaemia with the chromosomal translocation t(11;14)(p15;q11). AB - The ectopic expression of LMO1 or LMO2 in T cell acute leukaemias resulting from chromosomal translocations t(11;14)(p15;qll) or t(11;14)(p13;q11) respectively in a causal factor in tumorigenesis. LMO1 has been found as a heterodimer with a 46 Kd protein in a T cell line derived from a childhood T-acute leukaemia. This 46 Kd protein is the LIM-binding protein LDB1/NLI. The latter is a phosphoprotein and binds to LMO1 in its phosphorylated state and essentially all the LMO1 and LDB1 protein in the T cell line is part of the complex. Therefore, the LMO1-LDB1 interaction is likely to be involved in tumorigenesis after LMO1 is ectopically expressed following chromosomal translocation in T cells prior to development of acute leukaemias. PMID- 9872338 TI - Brain uptake of dihydroergotamine after intravenous and nasal administration in the rat. AB - This study was conducted to determine the uptake of dihydroergotamine (DHE) into the brain after intravenous and intranasal administration in rats. Eight anesthetized rats received either an intravenous (i.v.) or two successive intranasal (i.n.) doses of tritium labeled dihydroergotamine (3H-DHE) with 14C inulin as a non-BBB (blood-brain barrier) permeable marker. Radioactivity concentrations in plasma were determined at designated times within 30 min postdose, and in blood and seven brain regions (olfactory bulb, frontal cortex, parietal cortex, occipital cortex, cerebellum, mid-brain areas, and brain stem) at 30 min. The plasma-to-brain permeability*area product (PeA) following an i.v. dose was calculated based on the 30-min brain tissue concentration and the area under the plasma concentration-time curve (AUC0-30 min, i.v.) assuming unidirectional transport from plasma to brain. Direct transport from nasal cavity to brain was assessed based on the amount of radioactivity in brain determined experimentally and predicted based on plasma AUC0-30 min, i.n. and PeA obtained from i.v. data. Following an i.v. dose, DHE distributed into the brain with a brain-to-plasma concentration ratio of approximately 5% at 30 min postdose. The PeA value of DHE ranged from 8.6 x 10(-4) to 37.5 x 10(-4) mL min(-1) g(-1) in different brain regions. Following i.n. doses the experimentally determined concentration in olfactory bulb was approximately 51 times, and in other regions three to seven times, greater than predicted values based only on PeA and plasma AUC, suggesting a direct transport pathway from the nasal cavity to the brain. As a result, the brain tissue concentrations at 30 min were similar to (0.31-1.04 times) those following an i.v. dose except for the olfactory bulb, in which the concentration was approximately four times greater than that following an i.v. dose. In conclusion, 3H-DHE penetrated the BBB following intravenous administration. Following i.n. doses, 3H-DHE was able to enter the brain directly from the nasal cavity, with the olfactory bulb being a part of the direct passage from nasal cavity to brain. PMID- 9872337 TI - Transdermal absorption of clindamycin and tretinoin from topically applied anti acne formulations in man. AB - The percutaneous absorption of clindamycin was studied in healthy male volunteers, comparing two investigative clindamycin (% w/v)/tretinoin (0.025% w/v) gels, containing clindamycin phosphate ester and clindamycin HCl, respectively, relative to a clindamycin phosphate lotion (1% clindamycin; Dalacin T). Formulations were applied daily for 5 days on the face, according to a balanced complete block design. Redness of the skin was scored visually, and blood and urine were collected. Clindamycin plasma levels did not exceed the limit of quantification (5 ng mL(-1)) with the clindamycin phosphate formulations, but one volunteer who received the clindamycin HCl/tretinoin gel showed plasma levels of up to 13 ng mL(-1). Clindamycin urinary excretion for 12 h after application of the clindamycin phosphate/tretinoin gel was comparable to the values of the reference lotion, whereas the clindamycin HCl/tretinoin gel gave significantly higher values. Erythema appeared to be associated with increased urinary excretion. The formulations were tolerated well. In a separate clinical pilot study in acne patients, the transdermal uptake of tretinoin and clindamycin from the clindamycin phosphate/tretinoin gel was monitored. Plasma samples were collected after 4 and 12 weeks of daily treatment. None of the study plasma samples contained measurable tretinoin levels. Clindamycin levels were not quantifiable in the majority (87%) of samples, the highest plasma level was 11 ng mL(-1). The chemical form of clindamycin proved to modulate skin irritation and percutaneous uptake of clindamycin from a gel formulation in healthy subjects. There was no indications for a notable transdermal uptake of tretinoin during daily application of the gel in patients, nor for an enhancing effect of tretinoin on clindamycin uptake. PMID- 9872339 TI - Pharmacokinetics and tolerability of intravenous rizatriptan in healthy females. AB - The pharmacokinetics and tolerability of intravenous (i.v.) rizatriptan (MK 0462), a novel 5-HT1D/1B receptor agonist for the acute oral treatment of migraine, were examined in an open, single-dose, four-period, randomized crossover study in healthy females. Results of this study indicated that i.v. rizatriptan (0.5-5 mg) was well tolerated. The disposition kinetics of rizatriptan were linear for i.v. doses up to and including 2.5 mg. Relative to the 0.5 mg dose, geometric mean dose-adjusted AUC ratios were 1.04, 1.09, and 1.18 for 1, 2.5, and 5 mg doses, respectively. Apparent plasma clearance (Cl) ranged between 859 and 941 mL min(-1) from 0.5 to 2.5 mg, but dropped to slightly below 800 mL min(-1) for the 5 mg dose. Therefore, the elimination of rizatriptan appears somewhat dose dependent at the high end of this dose range. Mean plasma half-life (t1/2) was 1.5-2.2 h across all doses while mean residence time in the body (MRT) and steady state volume of distribution (Vss) of rizatriptan remained relatively invariant across doses. Urinary excretion of rizatriptan (Ue) ranged from 14.5 to 34.6% of dose. PMID- 9872340 TI - Single dose pharmacokinetics of HEPP, a new anticonvulsant in normal healthy volunteers. AB - The pharmacokinetics and the dose proportionality of a new anticonvulsant compound, HEPP (D,L-3-hydroxy-3-ethyl-3-phenylpropionamide) was studied in healthy male volunteers as part of the pharmacological evaluation for new drugs. Study was performed administering doses of 250, 375, 500 and 625 mg of HEPP to six male volunteers. Blood and urine samples were collected for 72 h postdose and analysed by HPLC. Results showed that in man HEPP is rapidly absorbed from the gastrointestinal tract. Tmax values were between 1.5 and 6.0 h. Plasma mean terminal half-life after the different doses ranged between 15.83 and 27.62 h with an overall harmonic mean value of 22.8. The mean AUC0-infinity and Cmax increased linearly with doses of 250, 375 and 500 mg but not with the dose of 625 mg. The amount of unchanged drug excreted in urine was between 3 and 6% of administered dose which shows an extensive metabolism of the drug. PMID- 9872341 TI - Avoidance of hepatic first-pass effect in the rabbit via rectal route of administration. AB - To assess avoidance of hepatic first-pass effect of drugs, we undertook in situ experiments using rectal administration of lidocaine in the rabbit. We also employed in situ duodenal route to estimate first-pass metabolism across the gastrointestinal mucosa. Rabbits were administered lidocaine HCl intravenously (i.v., 50 mg/20 min) and portally (i.p.v., 33.3, 16.7, 8.3 mg/20 min) and avoidance of hepatic first-pass effect (Fh) was calculated from the area under curve (AUC). Fh was about 30% and did not vary with increasing i.p.v. dose. Intravenous and i.p.v. administration was followed by duodenal (i.d.) or rectal (i.r.) administration and the absorption (fa), Fh, and avoidance of first-pass effect in the duodenal mucosal membrane (Fm) were determined. With i.d. administration, lidocaine was absorbed completely with negligible first-pass effect in the mucosa (Fm=1). On the other hand, while lidocaine was also absorbed almost completely via the i.r. route, avoidance of first-pass effect was 60%, representing twice the bioavailability via i.d. administration. On the basis of these data, assuming that the first-pass effect in the rectal mucosa was negligible, we estimate the fraction of rectal venous drainage bypassing the portal circulation and thus hepatic metabolism (fnh) to be about 40%. PMID- 9872342 TI - Stability of finite difference deconvolution II: simulation studies. AB - Theoretical analysis of the stability of finite difference deconvolution (FDD) indicates that if the cumulative amount function is used to characterize the drug input the method is stable for any sampling schedule for an intravenous unit impulse response function. The analysis also indicates that the method is stable for an oral unit impulse response only for well designed sampling schedules. This article confirms these results through numerical simulation experiments. It is shown that the assumption that the unit impulse response is error-free has an influence on the performance of FDD which is generally of no practical significance, except possibly for the first few points estimated. It is also shown that there is no significant interaction between the statistical error due to data noise and the deterministic algorithm error. The major source of error in practice is likely to be the data noise in the input response function. The simulations confirm that, with the estimated cumulative amount function as the quantity estimated and, with a well designed sampling schedule for the case of an oral unit impulse response, FDD is in practice an accurate and stable method with acceptable precision under a typical error disturbance. PMID- 9872343 TI - Intestinal absorption and analgesic activity of aminopeptidase-resistant cellobiose-coupled leucine enkephalinamide. AB - Leucine enkephalinamide (LEamide), aminopeptidase-degradable opioid peptide, was coupled with cellobiose (cellobiose-coupled LEamide, CcpLEamide). CcpLEamide was absorbed from the rat small intestine in vitro, whereas LEamide was not. CcpLEamide on the mucosal side was more stable than aminopeptidase-resistant cellobiose-coupled leucine enkephalin (CcpLE) in the presence of inhibitors of enkephalinase and angiotensin converting enzyme, and much more stable than LEamide. The absorption rate (clearance) of CcpLEamide was comparable with that of CcpLE in the presence of these peptidase inhibitors. Analgesic activities of these enkephalins were tested by the acetic acid writhing assay and hot-plate assay after subcutaneous administration to mice. Both assays indicated CcpLEamide induced analgesia. On the other hand, the analgesic activity of LEamide was not observed, but pretreatment with amastatin (a peptidase inhibitor) produced LEamide-induced analgesia. These results indicate that CcpLEamide is stable in the body and has analgesic effects without pretreatment with peptidase inhibitors, and was stable enough to be absorbed from the small intestine. We propose CcpLEamide as an orally active analgesic peptide candidate. PMID- 9872344 TI - Benzodiazepine treatment for alcohol-dependent patients. AB - Benzodiazepines (BZDs) are the preferred pharmacological agents for treatment of acute alcohol withdrawal. Treatment with BZDs can be administered on an out patient basis for subjects experiencing mild to moderate withdrawal and on an in patient basis for the most severe forms of withdrawal. The efficacy of BZDs for long-term treatment of alcoholism has been more controversial. Controlled studies indicate that BZD treatment does not improve abstinence rate. Most reviews of drug treatment of alcoholism conclude that routine use of BZDs is not indicated on a long-term basis. However, the clinical reality is that many alcoholics are treated by BZDs during detoxification and then continue to receive them for the treatment of anxiety disorders or insomnia, often secondary to alcohol dependence. After a review of the biological properties of BZDs related to their therapeutic issues, this review discusses the major indications for BZD treatment of alcoholism. BZDs are first prescribed to prevent and treat symptoms of alcohol withdrawal. Indication of BZD administration during alcohol withdrawal and criteria of choice of an agent according to its half-life or its route of administration are discussed. The different protocols of BZD treatment during withdrawal are considered (e.g. loading techniques, symptom-triggered therapy). The use of BZDs in the treatment of anxiety associated with alcohol dependence is examined. Among unwanted effects, risk of abuse, memory impairment, confusion, and delirium are described. Finally, practical guidelines for the use of BZDs in the treatment of alcoholism are proposed. PMID- 9872345 TI - Alcohol, health, and the heart: implications for clinicians. AB - The association between light drinking and reduced mortality compared to lifetime abstinence is robust. Confounders, such as unhealthy diet, social class, and social isolation, which are associated both with abstaining and with poorer health, have made it difficult for doctors to assess the meaning of the association, and to know how to respond to individual patients seeking advice on 'drinking for health'. This paper attempts to bring up to date the evidence concerning the confounders. The evidence for a preventive effect is reviewed according to pattern of alcohol use and type of patient. The balance of risk to benefit appears to favour giving medical advice to some patients in middle life with uncontrollable risk factors for coronary heart disease (and probably of ischaemic stroke), who are very infrequent drinkers, to increase slightly the frequency of drinking (not the per session amount) and for some abstainers to consider starting to take alcohol. However, caveats render quite small the number of patients to whom physicians will give this advice. There is also the theoretical risk of a ripple in the population such that more people may move into harmful drinking. PMID- 9872346 TI - Sexual abuse and the subsequent development of alcohol problems. AB - The literature on sexual abuse and alcohol problems has been reviewed. Various methodological issues are relevant in determining whether there is merely an association or also a causal relationship. These include the definition of sexual abuse, the degree and timing of abuse, the methods of data collection, sample selection, the presence or absence of control groups, possible recall bias, difficulties with prospective studies for this subject, and the definition of alcohol misuse or dependence. Results with community and victim samples are conflicting, but studies on samples of problem drinkers suggest an association between severe alcohol problems and previous sexual abuse, at least in women. The association may be especially strong for earlier and more severe forms of sexual abuse. Possible mechanisms for an association were examined and are: (1) sexual abuse as a cause of alcohol misuse; (2) alcohol misuse predisposing people to sexual assault; (3) sexual assault and alcohol misuse both resulting from another factor; (4) sexual abuse predisposing to other conditions associated with alcohol misuse; and (5) an artefactual association. Regardless of the role of sexual abuse in causing alcohol problems, the available evidence suggests that victims of sexual abuse may present to services with more problematical patterns of drinking and more concurrent psychiatric disorder. PMID- 9872347 TI - Improved autonomic neurocardial balance in short-term abstinent alcoholics treated with acamprosate. AB - Standardized investigations on resting heart rate variability (HRV) should provide more information on acamprosate's human pharmacodynamic properties because acamprosate interacts with several neurotransmitter systems which are also involved in maintaining autonomic neurocardiac balance. We performed HRV measurements prospectively in 69 healthy controls and 19 chronic alcoholics to prove the hypotheses that: (1) compared to healthy controls, chronic alcoholics show disturbances in neurocardiac vagal function; and (2) in alcoholics, acamprosate treatment (6-8 days) should further decrease parasympathetic activity if acamprosate interacts with central gamma-aminobutyric acidA receptors in vivo. Cardiovagal dysfunction was initially present in 21% of the alcoholics. After treatment. however, their neurocardiac sympathetic-parasympathetic balance improved significantly. PMID- 9872348 TI - Presence of plasma endotoxin is correlated with tumour necrosis factor receptor levels and disease activity in alcoholic cirrhosis. AB - Cytokines and plasma endotoxin were measured in a consecutive series of patients with alcoholic cirrhosis (AC). Endotoxaemia was found to be strongly correlated to increased plasma levels of functionally active tumour necrosis factor (TNF) receptors -p55 and -p75, TNF-alpha and the Child-Pugh stage of the disease. Our data support the hypothesis of the pathogenic role of lipopolysaccharide in hepatocellular damage of patients with AC. PMID- 9872349 TI - The effects of the nitric oxide synthase inhibitor 7-nitroindazole on ethanol pharmacokinetics in rats after acute and chronic ethanol administration. AB - The aim of this work was to study the effects of the nitric oxide synthase (NOS) inhibitors 7-nitroindazole (7-NI) and NG-nitro-L-arginine (L-NOARG) on the effects and pharmacokinetics of ethanol in rats. Ethanol at a dose of 4 g/kg, i.p. induced sleep in rats (sleep time: 117.2+/-30.7 min). Administration of the NOS inhibitors 7-NI (20 mg/kg, i.p.) and L-NOARG (20 mg/kg, i.p.) 30 min before ethanol significantly increased the duration of ethanol-induced sleep. L-NOARG also significantly increased the toxicity of ethanol as evidenced by increased post-experimental lethality. Ethanol at a dose of 2 g/kg (i.p.) did not induce sleep in vehicle-treated rats; however, the combined administration of ethanol (2 g/kg) and 7-NI at doses of 40, 80, and 120 mg/kg caused sleep, for 49.4+/-3.7, 204.0+/-13.3, and 447.5+/-62.8 min, respectively. L-NOARG (20 mg/kg) had no effect on ethanol concentrations in blood after acute ethanol administration (4 g/kg). 7-NI in lower doses (20 and 40 mg/kg) had no effect and in higher doses (80 and 120 mg/kg) significantly slowed ethanol clearance during the 12 h after ethanol administration. The effect of 7-NI (20 mg/kg) on ethanol pharmacokinetics after chronic ethanol administration (inhalation for 18 days) was also studied. The administration of 7-NI immediately after the end of ethanol exposure had a pronounced effect on ethanol pharmacokinetics; in 7-NI-treated rats the fall in ethanol concentrations was significantly slower as compared with vehicle-treated rats. In 7-NI-treated rats, blood-ethanol levels were higher at 3, 6, 9, and 12 h after the end of ethanol exposure. PMID- 9872350 TI - Tryptophan metabolism in alcoholism. Tryptophan but not excitatory amino acid availability to the brain is increased before the appearance of the alcohol withdrawal syndrome in men. AB - Tryptophan (Trp) metabolism and disposition and excitatory and other amino acid concentrations were determined in alcohol-dependent subjects in relation to the alcohol-withdrawal syndrome (AWS). Parameters were examined in 12 alcohol dependent male subjects, undergoing elective upper digestive tract tumour resection, and 12 age-, gender-, and medication-matched controls on three occasions: pre-operatively, post-operatively, and immediately before (i.e. within 24 h of) the appearance of the AWS. No significant differences were observed between controls and alcoholic subjects on the first or second of these occasions. On the third occasion, within 24 h of the appearance of the AWS, alcoholics showed a dramatic elevation (117%) in free serum Trp concentration and a consequent increase (111%) in the ratio of [free Trp]/[competing amino acids], which is an accurate predictor of Trp entry into the brain. Increases were also observed on this third occasion in concentrations of total Trp (49%), cortisol (123%), and norharman (137%). Concentrations of glutamate, glycine, aspartate, serine, and taurine did not differ significantly within or between the control and alcohol-dependent groups of subjects on any of the three occasions. The possible significance of the Trp and related metabolic changes in relation to the behavioural features of the AWS is discussed. PMID- 9872351 TI - Limitations in the use of gamma-glutamyl transferase estimations in alcohol dependent subjects. AB - This paper studies a cohort of randomly selected attenders at a district alcohol treatment service and examines the relationship between clinical assessments and laboratory markers currently in use in the unit. It measures in particular the rate of change of serial gamma-glutamyl transferase (GGT) during abstinence throughout an alcohol treatment programme in alcohol-dependent subjects. The results show that GGT is less often elevated in alcohol-dependent patients than was previously thought. Its predictive value changes little with respect to the age of the subject and length of drinking history. Measurement of GGT adds little to the diagnostic sensitivity of careful history taking. In the alcohol-dependent population, GGT estimation is of little value and a normal GGT does not exclude chronic alcohol dependence. PMID- 9872352 TI - Benfotiamine in treatment of alcoholic polyneuropathy: an 8-week randomized controlled study (BAP I Study). AB - A three-armed, randomized, multicentre, placebo-controlled double-blind study was used to examine the efficacy of benfotiamine vs a combination containing benfotiamine and vitamins B6 and B12 in out-patients with severe symptoms of alcoholic polyneuropathy (Benfotiamine in treatment of Alcoholic Polyneuropathy, BAP I). The study period was 8 weeks and 84 patients fulfilled all the prerequisite criteria and completed the study as planned. Benfotiamine led to significant improvement of alcoholic polyneuropathy. Vibration perception (measured at the tip of the great toe) significantly improved in the course of the study, as did motor function. and the overall score reflecting the entire range of symptoms of alcoholic polyneuropathy. A tendency toward improvement was evident for pain and co-ordination; no therapy-specific adverse effects were seen. PMID- 9872353 TI - CDTect-RIA and CDTect-EIA for determination of serum carbohydrate-deficient transferrin compared. AB - CDTect-RIA and CDTect-EIA for determination of serum carbohydrate-deficient transferrin (CDT) by radioimmunoassay and enzyme immunoassay respectively were tested for equality and precision in four European laboratories. For correlational studies, serum samples with CDT concentrations up to 130 U/l were analysed in accordance with a uniform trial schedule. The regression of CDT values obtained by the two procedures was computed for each laboratory using the method of Passing and Bablok. Slopes and intercepts of the regression functions did not differ significantly from the values 1 or 0, as proved by the corresponding 95% confidence intervals. Precision studies were computed using analysis of variance. For CDT concentrations at the upper reference limit for men, the within-day coefficients of variation (CVs) ranged between 0.7 and 6.4% (median 5.2%) for CDTect-RIA and from 4.3 to 9.2% (median 6.2%) for CDTect-EIA. The corresponding pure between-day CVs were 5.0-18.5% (median 9.8%) and 3.5-14.5% (median 10.9%). The study demonstrates the equality of CDT values obtained by CDTect-RIA and CDTect-EIA. According to this study, the two methods can be used interchangeably without getting fluctuating CDT values, e.g. in longitudinal studies. PMID- 9872355 TI - Intervention by an alcohol health worker in an accident and emergency department. AB - We have evaluated the effect of brief intervention by an alcohol health worker on alcohol consumption by 202 alcohol misusing patients. For 71 patients where questionnaires were successfully completed, 46 (65%) reported drinking less alcohol at 6 months as opposed to six (8.5%) who reported drinking more. The overall reduction was statistically significant (P < 0.001), with a mean reduction of 43%. PMID- 9872354 TI - Carbohydrate-deficient transferrin: marker of actual alcohol consumption or chronic alcohol misuse? AB - Carbohydrate-deficient transferrin (CDT) is a useful indicator of excessive alcohol consumption with higher sensitivity and specificity than other markers that are used. In the present study, CDT was analysed in 161 patients hospitalized in a surgical ward to evaluate whether history of drinking and chronic alcohol misuse are important determinants of CDT elevations. Fifty-one of the patients were diagnosed as alcohol-dependent and they all reported a long history of alcohol abuse. Several of these, as well as many of the non-dependent patients, reported a high, recent alcohol consumption (> or = 60 g/day for the previous 2 weeks). CDT performed better in detecting patients with alcohol dependency than in detecting patients with high alcohol consumption irrespective of dependency, showing higher sensitivity (47 vs 37%), likelihood ratio (4.7 vs 3.4), and a statistically significant difference in the receiver-operating characteristic curve areas (P = 0.04 in a two-tailed comparison test). In two subgroups, one with alcohol-dependent and one with non-dependent patients, consuming similar amounts of alcohol (range: 60-170 g/day), the sensitivity of CDT was 52 and 5%, respectively. We conclude that CDT is a better marker for patients with chronic alcohol misuse than as a marker for high actual alcohol consumption alone. PMID- 9872356 TI - Semi-automated carbohydrate-deficient transferrin in primary biliary cirrhosis: a pilot study. AB - Primary biliary cirrhosis (PBC) is one of the few non-alcohol induced liver pathologies which causes false positive results in the evaluation of carbohydrate deficient transferrin (CDT) for the diagnosis of alcohol misuse. This phenomenon has only been observed when using the CDTect assay (Pharmacia & Upjohn, Uppsala, Sweden). In this study, we evaluated CDT in female PBC patients (n = 14) by a new CDT procedure, the %CDT turbidimetric immunoassay (TIA, Axis Biochemicals, Oslo, Norway) using the isoelectric focusing/immunoblotting/laser densitometry (IEF/IB/LD, Specialty Laboratories, Santa Monica, CA, USA) procedure as the gold standard. One of the PBC patients tested CDT+ by IEF/IB/LD (cut-off >9 densitometry units, DU) and %CDT TIA (cut off >6%); one patient tested at the cut off point of the IEF/IB/LD and another one tested at the cut-off point of the %CDT TIA. Thus, unlike CDTect, the %CDT TIA is a procedure that produces few false positives in PBC. PMID- 9872357 TI - Elevated carbohydrate-deficient transferrin predicts prolonged intensive care unit stay in traumatized men. AB - Carbohydrate-deficient transferrin (CDT) is reported to have a higher specificity in alcoholism than conventional markers. As the morbidity and mortality rates amongst chronic alcoholics are raised following trauma, the objective was to investigate if CDT could be used to predict prolonged intensive care unit (ICU) stay and an increased morbidity in patients with multiple injuries admitted to the ICU. In this prospective double-blind study, 66 traumatized male patients were transferred to the ICU following admission via the emergency room and operative management. Blood samples for CDT determination were taken upon admission to the emergency room, the ICU and on days 2 and 4 following admission. The patients were allocated a priori to two groups: high CDT group (CDT >20 U/l on admission to the emergency room) and low CDT group (CDT < or = 20 U/l). CDT values were determined by microanion-exchange chromatography and radioimmunoassay. Thirty-six patients had an elevated CDT value on admission to the emergency room. The high CDT group had a significantly prolonged ICU stay (median high CDT group: 13 davs; median low CDT group: 5 days). Major intercurrent complications, such as alcohol-withdrawal syndrome, tracheobronchitis, pneumonia, pancreatitis, sepsis, and congestive heart failure, were significantly increased in the high CDT group. The increased risk of pneumonia in the high CDT group may be related to the significantly increased period of mechanical ventilation. As high CDT values were associated with an increased risk of intercurrent complications and a prolonged ICU stay, it seems reasonable to use CDT as a marker in intensifying research work into preventing alcoholism-associated complications. PMID- 9872358 TI - MUC1 and endometrial receptivity. AB - Mucins, best known as the major constituent of mucus, are a family of highly glycosylated, high molecular weight (> or = 200 kDa) glycoproteins present on the surface of human epithelial cells. MUC1 has the features of an integral membrane protein. It has an extracellular tandem repeat domain that forms the major part of the core protein, and results in a highly repetitive structure, which is extremely immunogenic. In the protein there is also a proteolytic cleavage site reported in the proximal extracellular domain. The secreted form of MUC1 lacks the cytoplasmic tail, but it is not clear whether this results from alternative splicing or proteolysis and release of the free extracellular domain. The locus of the MUC1 gene is on band 21 of the long arm of chromosome 1 (1q21). Anti adhesion properties of this mucin are probably the result of the unique structure of the molecule. In mouse uterine epithelium, the homologue MUC1 is regulated with reduced expression in the implantation period, but in humans, expression is high during the peri-implantation period. MUC1 may inhibit the interaction between trophoblast and apical epithelium adhesion molecules at the time of implantation, giving the possibility of forming a uterine barrier for implantation. PMID- 9872359 TI - The relationship between BcI2, Bax and p53: consequences for cell cycle progression and cell death. AB - Each cell is under constant surveillance to maintain the integrity of its genome. Genomic lesions in a cell must be repaired before the onset of DNA replication and cell division. In the scenario that the genomic lesion is not repairable, the damaged cells are disposed in an orderly manner known as programmed cell death or apoptosis. Apoptosis and cell cycle progression are two intimately linked phenomena. Uncontrollable cell proliferation perturbs the cellular homeostasis and this can lead to malignancies, as well as organ dysfunction and developmental abnormalities. The biological pathway controlling cell fate is sequentially organized at the molecular level. Recent studies have made important contributions in advancing our knowledge of the mechanisms of cell cycle control and apoptosis regulation. A oncogene-derived protein, Bcl2, confers negative control in the pathway of cellular suicide machinery. A Bcl2-homologous protein, Bax, promotes cell death by competing with Bcl2. While Bax-Bax homodimers act as apoptosis inducers, Bcl2-Bax heterodimer formation evokes a survival signal for the cells. Both Bcl2 and Bax are transcriptional targets for the tumour suppressor protein, p53, which induces cell cycle arrest or apoptosis in response to DNA damage. In all, the coordinate performance of these molecules is crucial for controlling life and death of a cell. PMID- 9872360 TI - The effect of intrauterine levonorgestrel use on the expression of c-JUN, oestrogen receptors, progesterone receptors and Ki-67 in human endometrium. AB - The major regulators of endometrial function are oestrogen and progesterone, which act through binding their nuclear receptors and by activating transcription of their target genes. Interactions between steroid receptors and transcription proteins, e.g. c-JUN/AP-1, can modulate steroid action at the transcriptional level. The 19-nortestosterone-derived progestin, levonorgestrel, is used for contraception, treatment of menorrhagia and for endometrial protection during hormone replacement therapy, but the signalling pathways of its action are totally undefined. We examined the effect of an intrauterine system, releasing 20 microg of levonorgestrel per 24 h (LNG-IUS), on immunoreactive oestrogen receptor, progesterone receptor, c-JUN and Ki-67 expression in 29 endometrial specimens, obtained from fertile women using the LNG-IUS for contraception. Moderate to strong immunostaining for oestrogen receptors was observed in the stromal cells in all specimens, in glandular epithelial cells in 26 cases and in flattened luminal epithelial cells in 17 specimens. Decidualized stromal cells showed no progesterone receptor immunoreactivity in 19 of the 29 specimens, and weak to moderate immunostaining in 10 cases. Luminal epithelial cells were negative for progesterone receptor in all samples. Intense nuclear staining for C JUN was observed in epithelial cells in 26 and in decidualized stromal cells in all 29 of the samples. In 16 samples, Ki-67 immunoreactivity was evaluated as weak to moderate in decidualized stroma, and in 13 samples absent. Our data demonstrate that intrauterine release of LNG maintains constant expression of C JUN and exerts progestational effects in the endometrium in the absence of progesterone receptors. In contrast, LNG-IUS inhibits several cellular responses to oestrogen despite the presence of endogenous oestrogen and oestrogen receptors. These data suggest that the progestational effects induced by progesterone and levonorgestrel are mediated through different signalling pathways. PMID- 9872361 TI - Analysis of Yq microdeletions in infertile males by PCR and DNA hybridization techniques. AB - Defects in spermatogenesis have been found associated with deletions of different portions of Y chromosome long arm (Yq), suggesting the presence of the azoospermia factor in the control of spermatogenesis. We studied 67 men with idiopathic azoospermia and severe oligozoospermia, cytogenetically normal, for the presence of microdeletions on Yq chromosome. By using polymerase chain reaction (PCR) and Southern blotting techniques we analysed the AZFa, AZFb and AZFc loci on Yq, where deletions have been associated with defects in spermatogenesis. Deletions of a portion of the Y chromosome were detected in five patients. Four of these patients shared deletions in distal Yq11 interval 6, including the DAZ gene, while one patient lacked loci in the proximal Yq11. Testicular histology of two patients bearing distal Yq11 deletions showed two different spermatogenic defects including Sertoli cell-only (SCO) syndrome and maturation arrest, while the patient with microdeletions in the proximal Yq11 showed a SCO phenotype. PMID- 9872362 TI - Gamma-tubulin during the differentiation of spermatozoa in various mammals and man. AB - The distribution of gamma-tubulin as a marker of microtubule organizing centres (MTOC) was studied during spermiogenesis in rodents and in rabbit, monkey and man. A polyclonal antibody directed against human gamma-tubulin was used both for indirect immunofluorescence (IIF) and post-embedding immunogold procedures. In all species, gamma-tubulin was detected in the proximal and distal centrioles of round spermatids. In elongating spermatids, gamma-tubulin was predominantly found in the pericentriolar material (PCM) of both centrioles and particularly around the adjunct of the proximal centriole. At this level, some labelling was also associated with manchette microtubules, but other parts of the manchette and the nuclear ring were never labelled. We propose a role for distal centriole gamma tubulin in axoneme nucleation and centriolar adjunct gamma-tubulin in manchette nucleation. The disappearance of gamma-tubulin in mature spermatozoa indicates that sperm aster nucleation should be dependent on oocyte gamma-tubulin. Remnants of gamma-tubulin in some human spermatozoa suggest that paternal gamma-tubulin also could contribute to sperm aster formation. PMID- 9872363 TI - Fluorescent study of chromatin and tubulin in apparently unfertilized human oocytes following ICSI. AB - In this study we examined 138 oocytes which were meiotically mature and, on light microscopic examination, contained either no or one pronucleus following intracytoplasmic sperm injection (ICSI). Oocytes were fixed and simultaneously stained for chromatin (Hoechst 33258) and the spindle (alpha-tubulin antibody). In nine oocytes, no sperm nucleus was observed. The remaining oocytes were separated into two groups following staining; (i) oocytes which had remained at metaphase II after ICSI (n = 74); and (ii) oocytes in which resumption of meiosis was observed after ICSI (n = 55). In all oocytes in which sperm chromatin was absent no resumption of meiosis had occurred and therefore parthenogenetic activation by the process of ICSI seems to be a rare event. In 17 out of 74 (23%) oocytes which remained at metaphase II, staining identified premature chromosome condensation (PCC) of the sperm chromatin (G1-PCC). Sperm nuclear decondensation or further transformation of the sperm chromatin was observed in 56 out of 74 (76%) oocytes which remained at metaphase II after ICSI and in 46 out of 55 (84%) oocytes which had resumed meiosis, indicating that initiation of sperm decondensation is independent of the resumption of meiosis in the oocyte. In contrast, transition of the sperm nucleus beyond the decondensed stage only occurred in association with resumption of meiosis in the oocyte (no pronuclei in metaphase II oocytes). The presence of both male and female pronuclei in 53% of oocytes which had resumed meiosis indicates that changes in sperm chromatin beyond the initial decondensation stage are dependent on cytoplasmic conditions which also permit female pronuclear formation. PMID- 9872364 TI - Tyrosine phosphorylation of a 95 kDa protein and induction of the acrosome reaction in human spermatozoa by recombinant human zona pellucida glycoprotein 3. AB - Protein tyrosine phosphorylation and induction of the acrosome reaction (AR) in non-capacitated and capacitated human spermatozoa was investigated in response to recombinant human zona pellucida glycoprotein (rhZP3) produced by Chinese hamster ovary cells transfected with a plasmid containing human ZP3 cDNA. rhZP3 containing medium promoted the AR in a high proportion of capacitated spermatozoa (48.6 +/- 3.2%; P < 0.01) compared with control (no rhZP3) samples (14.8 +/- 2.1%). However, rhZP3-containing medium did not cause increased acrosomal exocytosis in non-capacitated spermatozoa (16.8 +/- 3.0%). Induction of the AR was associated with increased tyrosine phosphorylation of a 95 +/- 5 kDa epitope only in capacitated spermatozoa. A dose-dependent increase in the protein phosphorylation of a 95 kDa epitope in response to rhZP3 was detected by [gamma 32P]-ATP labelling of detergent-solubilized sperm proteins. When spermatozoa were co-incubated with monoclonal antibody 97.25 (mAb 97.25) recognizing a 95 kDa tyrosine kinase epitope, there was no rhZP3 induction of tyrosine phosphorylation of the 95 kDa protein. Such co-incubation also markedly inhibited the AR (23.9 +/ 3.1%). These results support the model that initial interaction of the fertilizing spermatozoon with ZP3 involves the tyrosine phosphorylation of a 95 kDa tyrosine kinase protein and that this requires capacitation. PMID- 9872365 TI - Cytogenetic analysis of spermatozoa in the father of a child with a de-novo reciprocal translocation t(7;9) (q22;p23). AB - Analysis of sperm chromosomes was carried out in the father of a child with a de novo reciprocal translocation t(7;9) (q22;p23) by G-banding and chromosome painting. Sperm metaphases were obtained using the zona-free hamster oocyte-human sperm fusion technique. A total of 138 complements were sequentially analysed by G-banding and fluorescence in-situ hybridization (FISH). The frequency of spermatozoa with structural chromosome abnormalities (5.1%) and the estimated conservative aneuploidy (1.4%) were within the range obtained in our control donors (6.9 and 4%). The sex ratio (45.3% X versus 54.7% Y) was not significantly different from the theoretical 1:1. A total of 309 sperm complements was analysed by FISH, 138 sequentially analysed by G-banding-FISH and another 171 analysed by FISH only. The frequencies of structural chromosome abnormalities for chromosomes 7 and 9 (0.6 and 0% respectively) were not significantly different from those obtained in our control donors (0.6 and 0.8%). No spermatozoa with the t(7;9) (q22;p23) were observed, showing no evidence for a germ-cell mosaicism. A statistically significant, positive association between sperm breakpoints and fragile sites (P = 0.0225) was observed. However, the coincidence between fragile sites and sperm breaks (80%) was not significantly different from that obtained in our control donors (79.2%). These results suggest that in this case the risk of structural chromosome abnormalities in further offspring is not increased, although an association between fragile sites and sperm chromosome breaks in the father does exist. PMID- 9872366 TI - Expression of intercellular adhesion molecule (ICAM)-1 mRNA and protein is enhanced in endometriosis versus endometrial stromal cells in culture. AB - The relationship between the immune and the endometrial systems has been recently suggested to be critical to the development of endometriosis. We previously showed that one of the molecules involved in the complex events that allow this interaction is the adhesion molecule intercellular adhesion molecule (ICAM)-1. This study was designed to evaluate whether differences in ICAM-1 mRNA and protein expression might exist between eutopic endometrial cells and ectopic cells derived from endometriomas. Stromal cells were dispersed from samples of endometrium and ovarian endometriomas biopsied synchronously from 24 patients with endometriosis. We established that the relative expression of ICAM transcript was significantly higher in ectopic cells than that found in cultures derived from endometrial samples. Moreover, ectopic cells demonstrated a significant overexpression of ICAM-1 protein in both its cell-bound and soluble form (sICAM-1) (P < 0.05). Interestingly, endometrial secretion of sICAM-1 was shown to vary during the menstrual cycle as proliferative phase samples released significantly higher concentrations of the soluble protein compared to the secretory phase. In contrast, this cycle-dependent pattern was absent in stromal cells derived from endometriomas. Moreover, interleukin (IL-1beta) was able to increase sICAM-1 shedding from endometrial cells in a concentration-dependent manner and this IL-1beta-mediated induction could be slightly enhanced by oestradiol. As sICAM-1 is able to interfere with ICAM-1-mediated immune functions, the release of higher concentrations from ectopic samples may be the mechanism by which ectopic endometrial cells escape immunosurveillance. PMID- 9872367 TI - Localization of apoptotic cells within the human endometrium and correlation between apoptosis and p21 expression. AB - The aim of the present study was to gain a better understanding of the localization of apoptotic cells within the human endometrium during the menstrual cycle and to elucidate the relationships among the following for the human endometrium: apoptosis, p21 expression, and cell proliferation. Apoptosis and p21 expression were identified mainly in the glandular cells of the basal layer in the late secretory phase. In contrast, cells positive for Ki-67 were observed predominantly in the functional layer (in the proliferative phase in glandular cells and in the secretory phase in stromal cells). A very strong positive correlation (r = 0.81; P < 0.001) was demonstrated between the number of apoptotic cells and the number of p21-positive cells present among the glandular cells but, topographically, individual apoptotic cells were not coincident with p21-positive cells in serial sections. The results of this study suggest that the proliferation of the glandular cells of the basal layer is regulated by both apoptosis and p21 expression, particularly in the late secretory phase. Such regulation may be necessary to maintain a healthy population of glandular cells in the basal layer of the endometrium. PMID- 9872368 TI - Differential response of basal and tetrahydrobiopterin-stimulated activities of placental type III nitric oxide synthase to sodium dodecyl sulphate: relation to dimeric structure. AB - The major enzyme isoform that synthesizes nitric oxide (NO) in first trimester human placentae is endothelial or type III NO-synthase (NOS III) which exhibits high specific activity in the microsomal fraction. In the present study, we investigated the possible protective and enzyme-stabilizing role of tetrahydropterin (BH4). The anionic detergent, sodium dodecyl sulphate (SDS) and thermal stress (freeze-thaw) were used as non-specific 'subunit-dissociating' agents, and alterations in enzyme activity and subunit structure were investigated. SDS (> or =0.05% w/v) resulted in significant inhibition both of basal and BH4-stimulated activities of NOS III, but the latter responded more sensitively. Preincubation of microsomes with SDS (> or =0.1%, w/v), followed by incubation in an SDS-depleted reaction mixture led to an inhibition of BH4 stimulated enzyme activity, while no change in the basal activity was noted. This indicated that the SDS effect is only fully reversible in the case of basal activity. Considering that basal activity is due to the presence of endogenous BH4 tightly bound to the enzyme, this differential sensitivity of basal and BH4 stimulated enzyme activities to SDS may be related to a putative differential protective effect of BH4 on the two subunits of the NOS III dimer. Western blot analysis revealed that the SDS-induced inhibition of enzyme activity could not be ascribed to disruption of the dimeric structure. This finding confirms the view that SDS may affect NOS III activity without necessarily deteriorating quaternary protein structure. Nevertheless, BH4 is essential in maintaining dimeric structure under denaturing conditions, e.g. SDS treatment and freezing/thawing; it is even able to reverse the dissociation caused by SDS. A model describing the interaction between BH4 and NOS III, and its implications on the physiology and pathology of the human placenta, is discussed. PMID- 9872369 TI - Virtual planning in patients with frontal lobe lesions. AB - The VIrtual Planning Test (VIP) was developed to investigate the planning and organisational abilities of 25 patients with frontal lobe neurosurgical lesions, comparing their performance with 25 matched control subjects. The task, presented in the form of a board game, simulates planning and execution of a set of target activities that have to take place over a four day period, involving those that are either preparatory for a fictional "trip" abroad, or those that relate to the subject's current environment (context). The frontal lobe patients were impaired overall, tending to select inappropriate activities associated with their current context and showing greater impairment on subtasks requiring only one, rather than two, preparatory acts. The findings are discussed in terms of the validity of the VIP test in identifying executive functioning impairment and the degree to which the pattern of deficits are consistent with neuropsychological models of executive function. PMID- 9872370 TI - Right-left orientation and significance of systematic reversal in children. AB - Shortly before the acquisition of right and left, which generally occurs around age 6-7 years, a very simple right/left discrimination task makes it possible to distinguish groups of children with strikingly different cognitive abilities. Preschool children aged from 5 to 6.4 years were asked to show their left hand, right eye, left ear and right hand. On a variety of simple cognitive tasks exploring verbal fluency, syntactic comprehension, working memory, visuo-spatial ability and number processing, children who made from 1 to 3 errors (14% of the sample) performed significantly worse than those who showed systematic reversal (30%) and those who made no error. Differential use of logical thinking can partially explain these differences. Neuropsychological implications of these developmental findings are discussed. PMID- 9872371 TI - Interhemispheric transfer in normals and acallosals: latency adjusted evoked potential averaging. AB - Interhemispheric transfer time (IHTT) can be estimated from visual evoked potentials (EPs). Latency adjusted averaging (LAA) produces EPs which have enhanced components. LAA also provides estimates of EP latency variance and signal-to-noise ratio (S/N). LAA was tested in analysis of EP-IHTT in normal and acallosal subjects. It was hypothesized that in normals S/N and latency variance would reveal signal degradation resulting from interhemispheric transfer. LAA in normals replicated IHTT findings for both P1 and N1 latency. Latency variance did not increase for cross-callosal measures, whereas the S/N measure showed significant EP degradation due to callosal transfer. EPs from five subjects with callosal absence (two commissurotomy; two complete and one partial callosal agenesis) showed significantly larger than normal latency variability, as well as decreased S/N ratios, for cross-hemisphere visual EPs. Results support the value of LAA in EP research on adequacy of hemispheric interactions in clinical populations. PMID- 9872372 TI - Methodology of motor skill and laterality: new test of hand preference in Macaca nemestrina. AB - Ten female pig-tailed macaques (Macaca nemestrina) were tested for hand preference and hand skill (i.e., speed of performance and error rate). The experimental task was naturalistic, calling for adaptive manual skills and fine manipulation: the monkeys had to remove small food rewards embedded in a vertical array, and precision opposition of thumb and forefinger was needed to extract each pellet. Each monkey was tested 10 times on 10 different days. The results indicated individual hand preference rather than population-level handedness; however, a tendency toward right predominance was found. The results on hand skill showed a relation between error rate and hand preference, as the preferred hand made fewer errors. A different and unexpected finding was obtained when skill was evaluated in terms of speed of performance: in adult subjects the left hand was quicker than the right. Therefore, different kinds of skill showed different patterns in relation to hand preference. PMID- 9872373 TI - Cortical distribution of EEG activity for component processes during mental rotation. AB - Alpha power (8-12 Hz) was monitored over the frontal, temporal, parietal and occipital lobes of the left and right cerebral hemispheres while participants mentally rotated three-dimensional shapes to match a specified target. By comparing the activational patterns generated during three experimental conditions, each designed to systematically isolate the involvement of the various subcomponents comprising this mental rotation task, it was suggested that the right frontal lobe mediates encoding and comparison/decision processes, while the left parietal and the left temporal region appear most involved in the generation of images and their mental rotation. A preliminary model describing the cooperative interaction of these cortical regions during mental rotation tasks is proposed. PMID- 9872374 TI - Evidence for separate allocentric and egocentric space processing in neglect patients. AB - Spatial orientation was investigated in two different conditions: (a) when the shape of the enclosure was the only available information; (b) when a clearly perceivable visual cue was added. Three groups of subjects were investigated: normal controls, right brain-damaged patients without and with hemispatial neglect. The performance of the first two groups clearly demonstrated the capacity to use the geometric properties of the environment and to integrate this information with an additional visual cue. Considered as a group, patients with hemispatial neglect were able to use the shape of the environment and, to a lesser extent, the additional visual cue. However, individual differences suggest two opposite performance patterns: two patients responded randomly when the shape of the environment was the only available information, and they improved considerably when the cue was offered; two other patients showed normal competence in dealing with the geometrical properties of the environment, but were unable to take advantage of the cue. The different lesion site in these two types of patients suggests a possible dissociation of processing based upon allocentric or egocentric coding of space in humans as well as in animals. PMID- 9872375 TI - Axial movements are relatively preserved with respect to limb movements in aphasic patients. AB - Apraxia is commonly manifested during the acute stage following left hemisphere cerebrovascular accident and typically co-occurs with aphasia. We examined 30 acute stroke patients with aphasia and apraxia in order to determine if such patients show evidence of preservation of selective subclasses of movements. Although Geschwind noted the preservation of axial movements to command in aphasic apraxic patients, his views were subsequently refuted. However, we found that aphasic apraxic patients of varying degrees of severity, including patients with global aphasia, showed relative preservation of axial movements to command and imitation. Theoretical interpretations and implications for acute neurologic rehabilitation are discussed. PMID- 9872376 TI - Unilateral spatial neglect in the late stage of Alzheimer's disease. AB - We report two patients with presumptive diagnosis of dementia of the Alzheimer's type (AD) who manifested a clear unilateral neglect syndrome in the late stage of the disease. At onset, their cognitive profiles were broadly similar to the usual cases of degenerative dementia, but, as the disease progressed, neglect appeared, becoming gradually more severe. MRI showed more pronounced cerebral atrophy in the right hemisphere in both patients and larger ventricles on the right in one of them. These two longitudinal single case studies show that neglect can occur in AD despite the insidious and progressive nature of this disease, and in spite of the fact that the underlying pathology affects the brain bilaterally. The appearance of neglect in the late stages of the patients' illness is interpreted as resulting from both the asymmetry in the atrophy and the disruption of compensatory mechanisms caused by the progressively worsening of the atrophy in the left hemisphere. PMID- 9872377 TI - Lateral asymmetries of pupillary responses. AB - We wanted to learn if pupillary changes induced by looking and attending to stimuli on the right and left are asymmetrical. In humans, there are hemispheric asymmetries in the control of attention-arousal systems. Because attention and arousal may influence pupil size, asymmetric pupillary responses may be seen when looking and attending in different directions. Twelve right-handed, healthy volunteers served as subjects. Using infrared pupillography, we recorded changes of pupillary diameter while subjects were looking and attending to the stimuli on the right and left sides of space. For the one second following a saccade, there are three phasic pupillary responses, an initial constriction (C1) then a dilation (D1), followed by constriction (C2). Evaluation of these three responses revealed right-left asymmetries with more pupil dilation (D1) when looking to the stimulus on the right. Our results suggest that subjects are more aroused when looking to the right than when looking to the left. PMID- 9872378 TI - An intermanual advantage for tactual matching. AB - Previous research reports both increased and decreased levels of performance for concurrent tactual tasks when they are carried out by one hand (intramanual condition) rather than shared between the hands (intermanual condition). Intramanual advantages have been attributed to the benefits of intrahemispheric processing which avoids the need for the transcallosal passage of information. Intermanual advantages are attributed to the benefits of sharing the task load between the cerebral hemispheres. The effects of inter- and intramanual activity was investigated in a group of 72 dextrals. Subjects were required to make same/different discriminations for a tactual matching task. Faster completion times were observed for the intermanual condition compared to the intramanual condition. This suggests that the task was carried out more efficiently when the hemispheres shared the processing load. The intermanual advantage was accentuated for judgements of difference which place more strain on processing resources. There was no evidence of a left hand advantage for tactual matching of the type observed for Braille. PMID- 9872379 TI - Distinct frontal regions for processing sentence syntax and story grammar. AB - Time is a fundamental dimension of cognition. It is expressed in the sequential ordering of individual elements in a wide variety of activities such as language, motor control or in the broader domain of long range goal-directed actions. Several studies have shown the importance of the frontal lobes in sequencing information. The question addressed in this study is whether this brain region hosts a single supramodal sequence processor, or whether separate mechanisms are required for different kinds of temporally organised knowledge structures such as syntax and action knowledge. Here we show that so-called agrammatic patients, with lesions in Broca's area, ordered word groups correctly to form a logical sequence of actions but they were severely impaired when similar word groups had to be ordered as a syntactically well-formed sentence. The opposite performance was observed in patients with dorsolateral prefrontal lesions, that is, while their syntactic processing was intact at the sentence level, they demonstrated a pronounced deficit in producing temporally coherent sequences of actions. Anatomical reconstruction of lesions from brain scans revealed that the sentence and action grammar deficits involved distinct, non-overlapping sites within the frontal lobes. Finally, in a third group of patients whose lesions encompassed both Broca's area and the prefrontal cortex, the two types of deficits were found. We conclude that sequence processing is specific to knowledge domains and involves different networks within the frontal lobes. PMID- 9872380 TI - Phonetically cued word-fluency, gender differences and aging: a reappraisal. AB - Five-hundred and three normal subjects were given a phonetically cued word fluency task in order to investigate the controversial issue of the influence of gender and aging on this task. Subjects were requested to say in one minute all the words that occurred to them, beginning with a given letter (F, P and L). Besides the expected significance of education, we observed a female advantage, but not a significant decline in the performance with aging. However, the slope of the line expressing word fluency as a function of age was different between females and males, indicating a greater sensitivity to aging for males. We provide inner and outer tolerance limits for this test in the normal population and give a formula useful for adjusting the raw scores on the basis of gender and education. The results are discussed with reference to previous findings and some hypotheses about the origins of the female advantage in PWF. PMID- 9872381 TI - VHS domain marks a group of proteins involved in endocytosis and vesicular trafficking. AB - Endocytosis is driven by a mechanism which is characterized by an orderly congregation of a large number of proteins which effectuate, first, formation of a coated vesicles, second, pinching off the vesicle and, third, regulated transport. True to the nature of many other proteins involved in multimolecular complexes, also endocytosis-associated proteins, such as Eps15, clathrin and AP 2, are characterized by distinct domains which mediate the protein-protein interactions. We now report that a group of well-established endocytosis and/or vesicular trafficking proteins possess a VHS domain, a recently described domain with an unknown function. We suggest that in these proteins VHS serves as a membrane targeting domain which by its specific features together with FYVE, SH3 and/or TAM domains, which are also present in some VHS-containing proteins, is involved in the stage-specific assembly of the endocytic machinery. PMID- 9872382 TI - Structure, function and regulation of the vacuolar (H+)-ATPases. AB - The vacuolar (H+)-ATPases (or V-ATPases) function to acidify intracellular compartments in eukaryotic cells, playing an important role in such processes as receptor-mediated endocytosis, intracellular membrane traffic, protein degradation and coupled transport. V-ATPases in the plasma membrane of specialized cells also function in renal acidification, bone resorption and cytosolic pH maintenance. The V-ATPases are composed of two domains. The V1 domain is a 570-kDa peripheral complex composed of 8 subunits (subunits A-H) of molecular weight 70-13 kDa which is responsible for ATP hydrolysis. The V0 domain is a 260-kDa integral complex composed of 5 subunits (subunits a-d) which is responsible for proton translocation. The V-ATPases are structurally related to the F-ATPases which function in ATP synthesis. Biochemical and mutational studies have begun to reveal the function of individual subunits and residues in V-ATPase activity. A central question in this field is the mechanism of regulation of vacuolar acidification in vivo. Evidence has been obtained suggesting a number of possible mechanisms of regulating V-ATPase activity, including reversible dissociation of V1 and V0 domains, disulfide bond formation at the catalytic site and differential targeting of V-ATPases. Control of anion conductance may also function to regulate vacuolar pH. Because of the diversity of functions of V ATPases, cells most likely employ multiple mechanisms for controlling their activity. PMID- 9872383 TI - Recent studies of T7 RNA polymerase mechanism. AB - Bacteriophage T7 RNA polymerase (T7 RNAP) is known to be one of the simplest enzymes catalyzing RNA synthesis. In contrast to most RNA polymerases known, this enzyme consists of one subunit and is able to carry out transcription in the absence of additional protein factors. Owing to its molecular properties, the enzyme is widely used for synthesis of specific transcripts, as well as being a suitable model for studying the mechanisms of transcription. In this minireview the recent data on the structure and mechanism of T7 RNAP, including enzyme promoter interactions, principal stages of transcription, and the results of functional studies are discussed. PMID- 9872384 TI - Role of CFTR and anion exchanger in bicarbonate fluxes in C127 cell lines. AB - C127 cell lines transfected with wtCFTR, delta F508CFTR or vector were employed to determine HCO3- fluxes in the presence or absence of functional CFTR, using the pH-sensitive dye BCECF. Both cytosolic alkalinization and acidification were due to activity of anion exchanger and were similar in the three cell lines, indicating that expression of CFTR did not influence anion exchanger activity. In C127wt cells only, cAMP elevating agents significantly stimulated HCO3- fluxes, insensitive to the inhibitor of anion exchanger 4,4'-diisothiocyanate dihydrostilbene-2,2'-disulfonic acid, suggesting that activated CFTR directly mediates both HCO3- influx and efflux and therefore can contribute to intracellular and extracellular pH regulation. PMID- 9872385 TI - Knocking out a specific tRNA species within unfractionated Escherichia coli tRNA by using antisense (complementary) oligodeoxyribonucleotides. AB - Methods for the preparation of an Escherichia coli tRNA mixture lacking one or a few specific tRNA species can be the basis for future applications of cell-free protein synthesis. We demonstrate here that virtually a single tRNA species in a crude E. coli tRNA mixture can be knocked out by an antisense (complementary) oligodeoxyribonucleotide. One out of five oligomers complementary to tRNA(Asp) blocked the aspartylation almost completely, while minimally affecting the aminoacylation with other 13 amino acids tested. This 'knockout' tRNA behaved similarly to the untreated tRNA in a cell-free translation of an mRNA lacking Asp codons. PMID- 9872386 TI - Downregulation of fibronectin transcription in highly metastatic adenocarcinoma cells. AB - Silencing of fibronectin (FN) expression seems to be one of the key mechanisms underlying metastatic behaviour. An inverse correlation exists between FN expression levels and the metastatic potential of two related murine mammary adenocarcinomas, M3 and MM3. Primary cultures of M3 tumour, which is moderately metastatic to lung (40% incidence), show a conspicuous FN extracellular matrix (ECM) and high levels of FN mRNA, while primary cultures of the highly metastatic MM3 tumour (95% lung incidence) are negative for FN in immunofluorescence and show at least 40-fold lower levels of FN mRNA, only detectable by RT-PCR, with a different pattern of alternatively spliced EDI isoforms compared to M3 cells. We show that the FN promoter sequence is not altered in MM3 cells. Transfection experiments with CAT constructs indicate that silencing occurs at the transcriptional level, involving the 220-bp proximal promoter region. PMID- 9872387 TI - Modulation of interleukin-8 activity by gingipains from Porphyromonas gingivalis: implications for pathogenicity of periodontal disease. AB - Gingipains are the major cysteine proteinases synthesized by Porphyromonas gingivalis which, in soluble form, are able to initially convert IL-8 (77 amino acid residues) to a more potent species truncated at the amino terminus, followed by slow degradation and destruction of chemokine biological activity. In contrast, the same enzymes when associated with bacterial outer-membrane blebs (vesicles), instantly degrade this chemokine. This division of enhancing and inactivating activity between soluble and membrane-bound gingipains can cause the compartmentalization of pro- and anti-inflammatory reactions to distal and proximal positions from bacterial plaque, respectively, which may explain why, despite the massive neutrophil accumulation at periodontitis sites, there is no elimination of infection. PMID- 9872388 TI - Regulation of phospholipase D activity in synaptosomes permeabilized with Staphylococcus aureus alpha-toxin. AB - In order to investigate the regulation of presynaptic phospholipase D (PLD) activity by calcium and G proteins, we established a permeabilization procedure for rat cortical synaptosomes using Staphylococcus aureus alpha-toxin (30-100 microg/ml). In permeabilized synaptosomes, PLD activity was significantly stimulated when the concentration of free calcium was increased from 0.1 microM to 1 microM. This activation was inhibited in the presence of KN-62 (1 microM), an inhibitor of calcium/calmodulin-dependent kinase II (CaMKII), but not by the protein kinase C inhibitor, Ro 31-8220 (1-10 microM). Synaptosomal PLD activity was also stimulated in the presence of 1 microM GTPgammaS. When Rho proteins were inhibited by pretreatment of the synaptosomes with Clostridium difficile toxin B (TcdB; 1-10 ng/ml), the effect of GTPgammaS was significantly reduced; in contrast, brefeldin A (10-100 microM), an inhibitor of ARF activation, was ineffective. Calcium stimulation of PLD was inhibited by TcdB, but GTPgammaS dependent activation was insensitive to KN-62. We conclude that synaptosomal PLD is activated in a pathway which sequentially involves CaMKII and Rho proteins. PMID- 9872389 TI - Mutations at position 1122 in the catalytic domain of the mouse ras-specific guanine nucleotide exchange factor CDC25Mm originate both loss-of-function and gain-of-function proteins. AB - The role of two residues within the catalytic domain of CDC25Mm, a mouse ras specific guanine nucleotide exchange factor (GEF), was investigated by site directed mutagenesis. The function of the mutant proteins was tested in vivo in both a Saccharomyces cerevisiae cdc25 complementation assay and in a mammalian fos-luciferase assay, and in in vitro assays on human and yeast Ras proteins. Mutants CDC25Mm(E1048K) and CDC25Mm(S1122V) were shown to be (partly) inactive proteins, similar to their yeast homologs. Mutant CDC25Mm(S1122A) showed higher nucleotide exchange activity than the wild type protein on the basis of both in vitro and in vivo assays. Thus, alanine and valine substitutions at position 1122 within the GEF catalytic domain originate mutations with opposite biological properties, indicating an important role for position 1122 in GEF function. PMID- 9872390 TI - Negative charge at the protein kinase CK2 site enhances recognition of the SV40 large T-antigen NLS by importin: effect of conformation. AB - SV40 large tumor-antigen (T-ag) nuclear import is enhanced by the protein kinase CK2 (CK2) site (Ser111Ser112) flanking the nuclear localization sequence (NLS). Here we use site-directed mutagenesis to examine the influence of negative charge and conformation at the site on T-ag nuclear import and recognition by the NLS binding importin subunits. Negative charge through aspartic acid in place of Ser111 simulated CK2 phosphorylation in enhancing nuclear accumulation to levels well above those of proteins lacking a functional CK2 site. This was shown to be through enhancement of T-ag NLS recognition by importin using an ELISA-based assay. Asp112-substituted mutants containing proline at positions 109, 110 (wild type position) or 111 were compared to assess the role of conformation at the CK2 site. Maximal nuclear import of the protein with Pro109 was lower than that of the Pro110 derivative, with the Pro111 variant even lower, these differences also being attributable to effects on importin binding. All results indicate a correlation of the initial nuclear import rate with the importin binding affinity, demonstrating that NLS recognition by importin is a key rate determining step in nuclear import. PMID- 9872391 TI - Functional and structural features of gamma-zeathionins, a new class of sodium channel blockers. AB - Gamma1- and gamma2-zeathionins (gamma1-Z and gamma2-Z) are members of a family of small and basic peptides involved in plant protection. These plant defensins exhibit remarkable structural similarity to scorpion neurotoxins and insect defensins. In the present report, we used the whole-cell patch clamp technique to investigate the inhibition of the sodium current (I(Na)) by gamma1-Z and gamma2-Z in the GH3 cell line. Both gamma1-Z and gamma2-Z rapidly and reversibly inhibited I(Na) without changing the kinetics or voltage dependence of activation or inactivation. To our knowledge, this is the first example of a plant protein that inhibits the sodium channel. From structural comparisons with the mu-conotoxins, a family of peptides that block the sodium channel, we detected some similar features that could provide the basis of inhibition of sodium channels by gamma zeathionins. PMID- 9872392 TI - Identification and characterization of N-acetylglucosamine-6-O-sulfate-specific beta1,4-galactosyltransferase in human colorectal mucosa. AB - 6-Sulfo-sialyl Lewis X structure is attributable to recognition between lymphocytes and high endothelial venules. However, the biosynthetic pathway still remains unclear. We found that a beta-galactosyltransferase (betaGalT) in human colorectal mucosa preferentially acts on GlcNAc-6-O-sulfate (6S-GN). 6S GN:beta4GalT was partially purified by UDP-hexanolamine-Sepharose and asialo agalacto-ovomucin-Sepharose chromatographies. The optimum pH of this enzyme was found to be 6.5-7.5 and the Michaelis constants for 6S-GN and UDP-Gal were 0.43 mM and 16 microM, respectively. The enzymatic activity was dependent on divalent cations and the substrate specificity was not affected by alpha-lactalbumin. This is the first demonstration of the occurrence of 6S-GN:beta4GalT. PMID- 9872393 TI - Hyperthermophilic redox chemistry: a re-evaluation. AB - The redox chemistry of Pyrococcus furiosus rubredoxin and ferredoxin has been studied as a function of temperature in direct voltammetry and in EPR monitored bulk titrations. The Ems of both proteins, measured with direct voltammetry, have a normal (linear) temperature dependence and show no pH dependence. EPR monitoring is not a reliable method to determine the temperature dependence of the Em: upon rapid freezing the proteins take their conformation corresponding to the freezing point of the solution. PMID- 9872394 TI - Studies on the binding of wheat germ agglutinin (Triticum vulgaris) to O-glycans. AB - The binding profile of Triticum vulgaris (WGA, wheat germ) agglutinin to 23 O glycans (GalNAc alpha1-->Ser/Thr containing glycoproteins, GPs) was quantitated by the precipitin assay and its specific interactions with O-glycans were confirmed by the precipitin inhibition assay. Of the 28 glycoforms tested, six complex O-glycans (hog gastric mucins, one human blood group A active and two precursor cyst GPs) reacted strongly with WGA and completely precipitated the lectin added. All of the other human blood group A active O-glycans and human blood group precursor GPs also reacted well with the lectin and precipitated over two-thirds of the agglutinin used. They reacted 4-50 times stronger than N glycans (asialo-fetuin and asialo-human alpha1 acid GP). The binding of WGA to O glycans was inhibited by either p-NO2-phenyl alpha,betaGlcNAc or GalNAc. From these results, it is highly possible that cluster (multivalent) effects through the high density of weak inhibitory determinants on glycans, such as GalNAc alpha1-->Ser/Thr (Tn), GalNAc at the nonreducing terminal, GlcNAc beta1--> at the non-reducing end and/or as an internal residue, play important roles in precipitation, while the GlcNAc beta1-->4GlcNAc disaccharide may play a minor role in the precipitation of mammalian glycan-WGA complexes. PMID- 9872396 TI - The complete sequence of the mitochondrial genome of Saccharomyces cerevisiae. AB - The currently available yeast mitochondrial DNA (mtDNA) sequence is incomplete, contains many errors and is derived from several polymorphic strains. Here, we report that the mtDNA sequence of the strain used for nuclear genome sequencing assembles into a circular map of 85,779 bp which includes 10 kb of new sequence. We give a list of seven small hypothetical open reading frames (ORFs). Hot spots of point mutations are found in exons near the insertion sites of optional mobile group I intron-related sequences. Our data suggest that shuffling of mobile elements plays an important role in the remodelling of the yeast mitochondrial genome. PMID- 9872395 TI - Ouabain-sensitive H,K-ATPase: tissue-specific expression of the mammalian genes encoding the catalytic alpha subunit. AB - Human ATP1AL1 and corresponding genes of other mammals encode the catalytic alpha subunit of a non-gastric ouabain-sensitive H,K-ATPases, the ion pump presumably involved in maintenance of potassium homeostasis. The tissue specificity of the expression of these genes in different species has not been analyzed in detail. Here we report comparative RT-PCR screening of mouse, rat, rabbit, human, and dog tissues. Significant expression levels were observed in the skin, kidney and distal colon of all species (with the exception of the human colon). Analysis of rat urogenital organs also revealed strong expression in coagulating and preputial glands. Relatively lower expression levels were detected in many other tissues including brain, placenta and lung. In rabbit brain the expression was found to be specific to choroid plexus and cortex. Prominent similarity of tissue specific expression patterns indicates that animal and human non-gastric H,K ATPases are indeed products of homologous genes. This is also consistent with the high sequence similarity of non-gastric H,K-ATPases (including partial sequences of hitherto unknown cDNAs for mouse and dog proteins). PMID- 9872398 TI - Identification of regions in which positive selection may operate in S-RNase of Rosaceae: implication for S-allele-specific recognition sites in S-RNase. AB - A stylar S-RNase is associated with gametophytic self-incompatibility in the Rosaceae, Solanaceae, and Scrophulariaceae. This S-RNase is responsible for S allele-specific recognition in the self-incompatible reaction, but how it functions in specific discrimination is not clear. Window analysis of the numbers of synonymous (dS) and non-synonymous (dN) substitutions in rosaceous S-RNases detected four regions with an excess of dN over dS in which positive selection may operate (PS regions). The topology of the secondary structure of the S-RNases predicted by the PHD method is very similar to that of fungal RNase Rh whose tertiary structure is known. When the sequences of S-RNases are aligned with the sequence of RNase Rh based on the predicted secondary structures, the four PS regions correspond to two surface sites on the tertiary structure of RNase Rh. These findings suggest that in S-RNases the PS regions also form two sites and are candidates for the recognition sites for S-allele-specific discrimination. PMID- 9872397 TI - Selective conservation of an E-protein gene promoter during vertebrate evolution. AB - The murine E-protein gene ME1 encodes a non-tissue-specific, helix-loop-helix transcription factor that is associated with morphological development. ME1 gene expression is regulated by a TATA-less promoter that contains multiple Sp1 consensus elements, E-boxes, and a novel transcription initiation site. In this study, we compared DNA homologous to the ME1 promoter from vertebrate species ranging from frog to human. A region of striking sequence similarity was identified in a region corresponding to the ME1 transcription initiation site (ME1 Inr). Within this region, a poly d(A) tract and a 9-bp inverted repeat (5' GTCCGCCTG) were highly conserved in all species that were examined. Protein complexes that recognized these DNA elements were present among distant vertebrates (frog, chick, monkey and human), and were able to bend the ME1 Inr to a similar extent (approximately 60 degrees) as the previously described murine MBP alpha and MBP beta proteins. Collectively, these results suggest that an ME1 Inr-like element and its associated proteins functioned in an ancestral vertebrate more than 350 million years ago. PMID- 9872399 TI - The influence of phenobarbital on cytochromes and reactive oxygen species in erythropoietin producing HepG2 cells. AB - Light absorption photometry of HepG2 cells treated with phenobarbital for enhancing the content of cytochrome P-450 and the synthesis of erythropoietin revealed an influence on all cytochromes detectable in the wavelength range between 400 and 620 nm. No correlation was found between specific changes of cytochrome P-450 absorption and increased EPO synthesis as proposed earlier by Fandrey et al. (Life Sci. (1990) 47, 127-134). In the present study, however, the increased erythropoietin synthesis could be related to a decreased intracellular hydroxyl radical level described as crucial for the oxygen regulated gene expression (Kietzmann et al., Biochem. J. (1998) 335, 425-432; Porwol et al., Eur. J. Biochem. (1998) 256, 16-23). PMID- 9872400 TI - In the rat, tumor necrosis factor alpha administration results in an increase in both UCP2 and UCP3 mRNAs in skeletal muscle: a possible mechanism for cytokine induced thermogenesis? AB - Since the discovery of the new members of the UCP (uncoupling protein) family, UCP2 and UCP3, very few studies have dealt with the regulation of their expression. Bearing this in mind, administration of a single intravenous injection of TNF-alpha (100 microg/kg body weight) to rats resulted in a significant increase in UCP2 (242%) and UCP3 (113%) gene expression in skeletal muscle. The results suggest a possible role for UCP2 and UCP3 in the increase of energy expenditure associated with cytokine treatment. PMID- 9872401 TI - Eukaryotic mRNAs encoding abundant and scarce proteins are statistically dissimilar in many structural features. AB - It is well known that non-coding mRNA sequences are dissimilar in many structural features. For individual mRNAs correlations were found for some of these features and their translational efficiency. However, no systematic statistical analysis was undertaken to relate protein abundance and structural characteristics of mRNA encoding the given protein. We have demonstrated that structural and contextual features of eukaryotic mRNAs encoding high- and low-abundant proteins differ in the 5' untranslated regions (UTR). Statistically, 5' UTRs of low-expression mRNAs are longer, their guanine plus cytosine content is higher, they have a less optimal context of the translation initiation codons of the main open reading frames and contain more frequently upstream AUG than 5' UTRs of high-expression mRNAs. Apart from the differences in 5' UTRs, high-expression mRNAs contain stronger termination signals. Structural features of low- and high-expression mRNAs are likely to contribute to the yield of their protein products. PMID- 9872402 TI - Expression of a functional barley sucrose-fructan 6-fructosyltransferase in the methylotrophic yeast Pichia pastoris. AB - The cDNA encoding sucrose-fructan 6-fructosyltransferase (6-SFT) from barley (Hordeum vulgare) has been expressed in the methylotrophic yeast Pichia pastoris, using a translational fusion into vector pPICZ alphaC, containing the N-terminal signal sequence of Saccharomyces cerevisiae alpha-factor to allow entry into the secretory pathway. Transformed Pichia produced and secreted a functional 6-SFT which had characteristics similar to the barley enzyme, but had a pronounced additional 1-SST activity when incubated with sucrose. PMID- 9872404 TI - Random coil conformation of a Gly/Ala-rich insert in IkappaB alpha excludes structural stabilization as the mechanism for protection against proteasomal degradation. AB - Peptide segments of multiple glycine and alanine residues prevent the proteolytic degradation of ubiquitinated proteins by the proteasome. The structure of a Gly/Ala-rich insert in IkappaB alpha was probed by nuclear magnetic resonance (NMR) spectroscopy, comparing IkappaB alpha samples with and without Gly/Ala-rich insert. Narrow 1H-NMR resonances at chemical shifts indicative of random coil conformations were observed in the difference spectrum. circular dichroism (CD) measurements further confirm that the mechanism of protection against proteolytic degradation is not based on structural transition or stabilization caused by the Gly/Ala-rich segment. In addition, most of the N- and C-terminal residues outside the ankyrin repeats in wild-type IkappaB alpha were found to be flexibly disordered. PMID- 9872403 TI - A new intracellular serine protease inhibitor expressed in the rat pituitary gland complexes with granzyme B. AB - We have cloned a novel serpin (raPIT5a) from a rat pituitary cDNA library which is structurally related to members of the ovalbumin subfamily of serine protease inhibitors. This new cDNA encodes a 374-amino acid protein, designated raPIT5a. raPIT5a was expressed in specific cells in the intermediate and anterior lobes of the pituitary. Recombinant raPIT5a was not secreted suggesting raPIT5a functions to inhibit intracellular proteases. Recombinant raPIT5a formed an SDS-stable complex with human granzyme B, a serine protease which induces apoptosis by activating members of the caspase enzyme family. These data suggest raPIT5a may have a role in regulating granzyme B or related enzymes and apoptosis in the pituitary gland. PMID- 9872405 TI - Cardiac specific expression of the green fluorescent protein during early murine embryonic development. AB - We demonstrate the establishment of transgenic mice, where the expression of the green fluorescent protein (GFP) is under control of the human cardiac alpha-actin promoter. These mice display cardiac specific GFP expression already during early embryonic development. Prominent GFP fluorescence was observed at the earliest stage of the murine heart anlage (E8). Cardiomyocytes of different developmental stages proved GFP positive, but the intensity varied between cells. We further show that contractions of single GFP positive cardiomyocytes can be monitored within the intact embryo. At later stages of embryonic development, the skeletal musculature was also GFP positive, in line with the known expression pattern of cardiac alpha-actin. The tissue specific labeling of organs is a powerful new tool for embryological as well as functional investigations in vivo. PMID- 9872406 TI - Antibodies against type II phospholipase A2 prevent renal injury due to ischemia and reperfusion in rats. AB - This study was performed to determine the involvement of type II phospholipase A2 (PLA2-II) in renal injury caused by ischemia and reperfusion. Ischemia and reperfusion significantly elevated levels of blood urea nitrogen and serum creatinine in rats. These increases were significantly reduced by i.v. administration of rabbit IgG F(ab')2 fragments against rat PLA2-II. Increased levels of acid-stable PLA2 activity in the kidney were caused by ischemia and reperfusion, and were suppressed by administration of anti-PLA2-II F(ab')2. Increased levels of myeloperoxidase activity, a marker of neutrophil infiltration, in the kidney were also reduced after anti-PLA2-II F(ab')2 treatment. These results suggest that PLA2-II plays a pivotal role in pathogenesis of ischemia and reperfusion injury through induction of neutrophil infiltration. PMID- 9872407 TI - C isotopomer analysis of glutamate by heteronuclear multiple quantum coherence total correlation spectroscopy (HMQC-TOCSY). AB - 13C has become an important tracer isotope for studies of intermediary metabolism. Information about relative flux through pathways is encoded by the distribution of 13C isotopomers in an intermediate pool such as glutamate. This information is commonly decoded either by mass spectrometry or by measuring relative multiplet areas in a 13C NMR spectrum. We demonstrate here that groups of glutamate 13C isotopomers may be quantified by indirect detection of protons in a 2D HMQC-TOCSY NMR spectrum and that fitting of these data to a metabolic model provides an identical measure of the 13C fractional enrichment of acetyl CoA and relative anaplerotic flux to that given by direct 13C NMR analysis. The sensitivity gain provided by HMQC-TOCSY spectroscopy will allow an extension of 13C isotopomer analysis to tissue samples not amenable to direct 13C detection (approximately 10 mg soleus muscle) and to tissue metabolites other than glutamate that are typically present at lower concentrations. PMID- 9872408 TI - The product of the mammalian orthologue of the Saccharomyces cerevisiae HBS1 gene is phylogenetically related to eukaryotic release factor 3 (eRF3) but does not carry eRF3-like activity. AB - We describe here the cloning and sequencing of human and mouse cDNAs encoding a putative GTP binding protein. Sequence comparison shows that these cDNAs (named eRFS) are likely to represent the orthologues of the yeast Saccharomyces cerevisiae HBS1 gene and that the C-terminal domains of the encoded proteins share structural features with eukaryotic elongation factor eEF-1A and release factor 3 (eRF3) families. The phylogenetic analysis suggests that eRFS proteins and Hbs1p form a cluster of orthologous sequences branching with the eRF3 family. Nevertheless, in yeast, the human eRFS protein and Hbs1p do not complement eRF3/Sup35p thermosensitive mutation and do not interact with eRF1. PMID- 9872409 TI - Homomultimeric protease in the hyperthermophilic bacterium Thermotoga maritima has structural and amino acid sequence homology to bacteriocins in mesophilic bacteria. AB - A novel homomultimeric protease (> 669 kDa), based on 31 kDa subunits, was purified from cell extracts of the hyperthermophilic bacterium Thermotoga maritima. This protease exhibits activity toward chymotrypsin and trypsin substrates, optimally at 90 degrees C and pH 7.1, and has a half-life of 36 min at 95 degrees C. Transmission electron microscopy established that the protease consists of a large globular assembly which appears circular from the front view. The function of this protease in T. maritima remains unclear, although putative homologs include a 29 kDa antigen from Mycobacterium tuberculosis and a 31 kDa monomer of a high molecular weight bacteriocin produced by Brevibacterium linens [Valdes-Stauber, N. and Scherer, S. (1996) Appl. Environ. Microbiol. 62, 1283 1286]. The relationship of these mesophilic proteins to the T. maritima protease suggests that their antibacterial activity may involve elements of proteolysis, and raises the prospect for antimicrobial ecological strategies in hyperthermophilic niches. PMID- 9872410 TI - Proteasome-dependent degradation of oxidized proteins in MRC-5 fibroblasts. AB - Fibroblasts were exposed to various concentrations of hydrogen peroxide and the removal of oxidized proteins was followed by determining protein-bound carbonyls. Fibroblasts are able to increase the turnover of metabolically radiolabeled proteins after treatment with hydrogen peroxide. It was demonstrated for the first time, that the increased protein turnover was accompanied by a removal of protein-bound carbonyl groups. The proteasome-specific inhibitor lactacystin was able to inhibit the elimination of protein-bound carbonyl groups. Therefore, the key role of the proteasome in the degradation of oxidized proteins in fibroblasts could be demonstrated. PMID- 9872411 TI - Estrogen withdrawal-induced human breast cancer tumour regression in nude mice is prevented by Bcl-2. AB - We recently showed that estrogen induces expression of the anti-apoptotic protein, Bcl-2 in MCF-7 human breast cancer cells. Since estrogen-dependent breast tumours can regress following estrogen withdrawal, we hypothesized that stable Bcl-2 expression would prevent estrogen-withdrawal induced regression of MCF-7 tumours. We therefore established tumours in ovariectomized female nude mice implanted with an estrogen-release pellet using untransfected MCF-7 cells or MCF-7 cells stably transfected with a Bcl-2 cDNA sense or antisense expression vector. All tumours grew at similar rates indicating that Bcl-2 levels have no effect on tumour formation. After removal of the estrogen pellet, Bcl-2 antisense tumours and untransfected MCF-7 tumours regressed means of 49% and 52%, respectively, after estrogen pellet removal whereas Bcl-2 sense tumours were significantly stabilized. Regressing tumours displayed characteristics of apoptotic cells. These results show that Bcl-2 can prevent hormone-dependent breast tumour regression and are consistent with the notion that decreased Bcl-2 levels following estrogen withdrawal renders hormone-dependent breast tumour cells sensitive to apoptotic regression. PMID- 9872412 TI - CD-monitored redox titration of the Rieske Fe-S protein of Rhodobacter sphaeroides: pH dependence of the midpoint potential in isolated bc1 complex and in membranes. AB - The redox potential of the Rieske Fe-S protein has been investigated using circular dichroism (CD)-spectroscopy. The CD features characteristic of the purified bc1 complex and membranes of Rhodobacter sphaeroides were found in the region between 450 and 550 nm. The difference between reduced and oxidized CD spectra shows a negative band at about 500 nm with a half of width 30 nm that corresponds to the specific dichroic absorption of the reduced Rieske protein (Fee, J.A. et al. (1984) J. Biol. Chem. 259, 124-133; Degli Esposti, M. et al. (1987) Biochem. J. 241, 285-290; Rich, P.R. and Wiggins, T.E. (1992) Biochem. Soc. Trans. 20, 241S). It was found that the redox potential at pH 7.0 for the Rieske center in the isolated bc1 complex and in chromatophore membranes from the R-26 strain of Rh. sphaeroides is 300 +/- 5 mV. In chromatophores from the BC17C strain of Rh. sphaeroides, the Em value measured for the Rieske iron-sulfur protein (ISP) was higher (315 +/- 5 mV), but the presence of carotenoids made measurement less accurate. The Em varied with pH in the range above pH 7, and the pH dependence was well fit either by one pK at approximately 7.5 in the range of titration, or by two pK values, pK1 = 7.6 and pK2 = 9.8. Similar titrations and pK values were found for the Rieske Fe-S protein in the isolated bc1 complex and membranes from the R-26 strain of Rb. sphaeroides. The results are discussed in the context of the mechanism of quinol oxidation by the bc1 complex, and the role of the iron sulfur protein in formation of a reaction complex at the Qo-site. PMID- 9872413 TI - The role of the cysteine-rich region of the beta2 integrin subunit in the leukocyte function-associated antigen-1 (LFA-1, alphaLbeta2, CD11a/CD18) heterodimer formation and ligand binding. AB - The cysteine-rich region (CRR) of the beta2 integrin subunit was replaced by that of beta1 to give the chimera beta2NV1. Beta2NV1 can combine with alphaL to form a variant leukocyte-function-associated antigen (LFA)-1 on COS cell surface, suggesting that the specificity of the beta2 interaction with alphaL does not lie in the CRR. Unlike those expressing wild-type LFA-1, COS cells expressing alphaL beta2NV1 are constitutively active in intercellular adhesion molecule (ICAM)-1 adhesion. These results suggest that activation of LFA-1 involves the release of an intramolecular constraint, which is maintained, in part, by the authentic beta2 CRR. PMID- 9872414 TI - BCL-2 promotes migration and invasiveness of human glioma cells. AB - Malignant progression in gliomas is correlated with increased migratory capacity which involves metalloproteolytic activity. Here, we report that ectopic expression of BCL-2 in two malignant glioma sublines markedly promoted glioma cell migration from spheroids and invasion into Matrigel-coated membranes. Invasion of fetal rat-brain aggregates was enhanced by BCL-2. Zymography revealed activation of matrix metalloproteinase-2 (MMP-2) in BCL-2-expressing cells. BCL-2 expressing cells showed an increase in MMP-2/-3/-12 (LN-18), and MMP-9/-12 and cell surface urokinase-type plasminogen activator (u-PA) (LN-229) mRNA and a reduction in tissue inhibitors of metalloproteinases (TIMP)-2 mRNA (LN-229). Taken together, we propose a novel function for BCL-2 in the malignant phenotype of glioma cells, that is, to enhance migration and invasion by altering the expression of a set of metalloproteinases and their inhibitors. PMID- 9872415 TI - AtMSI4 and RbAp48 WD-40 repeat proteins bind metal ions. AB - The mammalian RbAp48 protein is the most extensively studied member of the conserved family of Msi1-like WD-40 repeat proteins, which are components of complexes involved in the assembly and modification of chromatin. We have isolated a plant homolog of RbAp48, AtMSI4. By metal affinity chromatography, zinc blotting and atomic absorption analysis, we demonstrate that purified recombinant RbAp48 and AtMSI4 proteins bind 3-4 metal ions per molecule of protein. Metal competition assays indicate a preference for zinc. Both N- and C terminal halves of RbAp48 and AtMSI4 display zinc binding activity, suggesting it is an intrinsic property of the propeller structures likely to be formed by these proteins. Metal binding might mediate and/or regulate protein-protein interactions which are functionally important in chromatin metabolism. PMID- 9872416 TI - Repression of enzymes of the pentose phosphate pathway by glucose in fission yeast. AB - We examine here the effect of carbon sources on the synthesis of the shunt pathway enzymes in the fission yeast Schizosaccharomyces pombe growing on a mixture of ethanol and glycerol. Delta-gluconolactone induces practically every one of these enzymes. Glucose in contrast tends to attenuate the synthesis of the majority of them. RNA analysis confirms that their induction and repression reflect changes in the levels of their transcripts. PMID- 9872417 TI - Different patterns of IL-1beta secretion, adhesion molecule expression and apoptosis induction in human endothelial cells treated with 7alpha-, 7beta hydroxycholesterol, or 7-ketocholesterol. AB - Among oxysterols oxidized at C7 (7alpha-, 7beta-hydroxycholesterol, and 7 ketocholesterol), 7beta-hydroxycholesterol and 7-ketocholesterol involved in the cytotoxicity of oxidized low density lipoproteins (LDL) are potent inducers of apoptosis. Here, we asked whether all oxysterols oxidized at C7 were able to trigger apoptosis, to stimulate interleukin (IL)-Ibeta and/or tumor necrosis factor (TNF)-alpha secretion, and to enhance adhesion molecule expression (intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin) on human umbilical venous endothelial cells (HUVECs). Only 7beta-hydroxycholesterol and 7-ketocholesterol were potent inducers of apoptosis and of IL-1beta secretion. TNF-alpha secretion was never detected. Depending on the oxysterol considered, various levels of ICAM-1, VCAM-1 and E selectin expression were observed. So, oxysterols oxidized at C7 differently injure and activate HUVECs, and the alpha- or beta-hydroxyl radical position plays a key role in apoptosis and IL-1beta secretion. PMID- 9872418 TI - The stereochemical course of the reaction mechanism of trehalose phosphorylase from Schizophyllum commune. AB - Phosphorolysis of alpha,alpha-trehalose catalyzed by trehalose phosphorylase from the basidiomycete Schizophyllum commune proceeds via net retention of anomeric configuration and yields alpha-D-glucose 1-phosphate and alpha-D-glucose as the products. In reverse reaction, only the alpha-anomers of D-glucose 1-phosphate and D-glucose are utilized as glucosyl donor and acceptor, respectively, and give exclusively the alpha,alpha-product. Trehalose phosphorylase converts alpha-D glucose 1-fluoride and phosphate into alpha-D-glucose 1-phosphate, a reaction requiring the stereospecific protonation of the glucosyl fluoride by a Bronsted acid. The results are discussed with regard to a plausible reaction mechanism of fungal trehalose phosphorylase. PMID- 9872419 TI - Rapid hyaluronan uptake is associated with enhanced motility: implications for an intracellular mode of action. AB - Texas red-labeled hyaluronan (TR-HA) is rapidly taken up in a CD44 independent manner into ras-transformed 10T1/2 fibroblasts, where it accumulates in both cell ruffles/lamellae, the perinuclear area, and the nucleus. HA does not accumulate in the cell ruffles/lamellae of parental 10T1/2 cells. Addition of HA to ras transformed cells promotes their random motility but has no effect on 10T1/2 cell motility. 10T1/2 cells can be modified to take up HA into cell ruffles by exposure to phorbol ester or direct microinjection of HA into cells. Both treatments significantly stimulate 10T1/2 cell motility. PMID- 9872421 TI - Expression and characterization of human PDEdelta and its Caenorhabditis elegans ortholog CEdelta. AB - Cyclic GMP phosphodiesterase (PDE) is rod photoreceptor disk membrane-associated via C-terminal lipid tails. PDEdelta, a recently identified subunit, was shown to disrupt PDE/membrane interaction under physiological conditions, without affecting PDE catalytic activity. We found that a PDEdelta ortholog from the eyeless nematode Caenorhabditis elegans (termed CEdelta) solubilizes bovine PDE in vitro with an EC50 very similar to PDEdelta. Immobilized PDEdelta and CEdelta both bind, in addition to bovine PDE, an N-terminal fragment of human retinitis pigmentosa GTPase regulator, but not rhodopsin kinase and Ran binding protein 1. The results suggest that PDEdelta and CEdelta may regulate membrane binding of a variety of proteins in photoreceptors and other tissues. PMID- 9872422 TI - Cochlear implant performance and electrically-evoked auditory brain-stem response characteristics. AB - OBJECTIVES: The purpose of this study was to find a correlation between cochlear implant performances in phoneme discrimination and activity of the brain-stem. METHODS: Electrically-evoked auditory brain-stem responses (EABRs) and speech recognition performances were measured in 17 patients implanted with an MXM Digisonic DX10 cochlear implant. Speech recognition performances without lip reading were tested using lists of isolated French words containing 3 phonemes. RESULTS: The results indicated statistically significant correlations between phoneme correct-identification scores and the following EABR variables: wave V latency, wave II-V latency interval and wave III-V latency interval. These results, indicate that up to about 48% of the variance in isolated word recognition without lip-reading can be accounted for by EABR variables. CONCLUSION: The quality of brain-stem functioning influences central processes in phoneme discrimination. PMID- 9872420 TI - Increased uncoupling protein-2 and -3 gene expressions in skeletal muscle of STZ induced diabetic rats. AB - Streptozotocin (STZ)-induced diabetic animals are vulnerable to cold stress. Uncoupling proteins (UCPs) play an important role in regulating thermogenesis. We investigated the gene expressions of UCPs in brown adipose tissue (BAT), white adipose tissue (WAT), liver and gastrocnemius muscle of STZ-diabetic rats using Northern blot. UCP-1, -2 and -3 mRNA expressions in BAT were all remarkably lower in STZ-diabetic rats than those in control rats. Both UCP-2 and -3 gene expressions in gastrocnemius muscle were substantially elevated in STZ-diabetic rats and insulin treatment restored UCP gene expressions to normal levels. These results suggest that in STZ-diabetic rats, the overexpression of UCP-2 and UCP-3 in skeletal muscle provides a defense against hypothermogenesis caused by decreased UCPs in BAT. PMID- 9872423 TI - Quantification and rejection of ocular artifacts in auditory evoked fields in schizophrenics. AB - RESULTS: In a magnetoencephalographic investigation of the auditory evoked field (AEF) in 17 schizophrenics and 17 controls, 37% of the schizophrenics and 12% of the controls showed eye artifacts in every second trial or even more frequently. In the uncorrected average fields, the ratio between the power of artifacts and the power of the magnetoencephalogram (MEG) exceeded the value of 0.1 for 48% of the schizophrenics and for 29% of the controls. Ocular artifacts biased the locations of equivalent current dipoles of the M100 component towards deeper positions. A regression algorithm for the correction of ocular artifacts in raw data and an identification technique of ocular artifacts based on the topography of transmission coefficients is described. CONCLUSIONS: A linear dependence of ocular artifacts in AEF on the electrooculogram (EOG) was confirmed. Possible errors introduced by the correction are discussed. Transmission coefficients should be calculated for several individual trials with the same type of artifact. Errors due to evoked potentials in the EOG were found to be comparable in amplitude to noise in the AEF. Examples of transmission coefficients from the EOG to the MEG are given. PMID- 9872424 TI - High frequency oscillations in early cortical somatosensory evoked potentials. AB - OBJECTIVE: To evaluate the characteristics of high frequency (HF) components of the early cortical somatosensory evoked potentials (SEPs). METHODS: We recorded 8 channel SEPs from the frontal and left centro-parietal scalp after right median nerve stimulation with a wide band-pass (0.5-2000 Hz) and digitized at 40 kHz sampling rate in 12 healthy subjects. HF components were analyzed after digital band-pass filtering (300-1000 Hz). The power spectrum was obtained by a maximum entropy method. RESULTS: HF oscillations (maximum power at 600-800 Hz) consisting of 5 to 8 peaks were discriminated from the preceding P14 far-field in all cases and their phases were reversed between the frontal and contralateral parietal regions. In addition, in subjects with a high amplitude central P22 potential in original wide-band recordings, a single HF oscillation with a maximum at the central region was present. Furthermore, this component showed no phase reversal over the centro-parietal area. CONCLUSION: We therefore conclude that HF oscillations are superimposed not only on the tangential N20-P20 but on the radial P22 potential, and are generated from both tangential (area 3b) and radial (area 1) current sources. PMID- 9872425 TI - Evidence for efferent effects on early components of the human auditory brain stem evoked potentials. AB - OBJECTIVES AND METHODS: Auditory brain-stem evoked potentials (ABEPs) were recorded from 10 normal hearing subjects in response to rarefaction clicks, presented at a rate of 11/s. Stimuli were binaurally symmetrical and isochronic at 75 dB peSPL or with interaural time disparities (ITDs) of +/-0.4 ms, or intensity disparities (IIDs) of +/-10 dB. Potentials were recorded from vertex neck, as well as from 3 orthonormally positioned differential derivations. The amplified potentials were averaged over 8000 repetitions using a dwell time of 20 micros/address/channel. The effects of contralateral stimulation on neural responses of the peripheral auditory system were obtained by subtracting the binaural response from the algebraic sum of responses to left and right monaural stimuli. From the 3 orthonormal derivations, 3-channel Lissajous' trajectories (3 CLTs) to the various stimulus conditions and difference waveforms were derived. RESULTS: The results corroborated earlier studies on binaural interaction components (BICs), which include 3 major components corresponding in latency to the vertex-mastoid peaks IV-VI of ABEP. In addition, the binaural difference waveforms included 3 earlier, low-amplitude components. Latency correspondence and comparison of difference waveform and ABEP 3-CLTs indicated that the first and third early difference waveform components corresponded to the negative peaks following I and III, respectively, of the vertex-neck ABEP to binaural clicks. CONCLUSIONS: These results indicate that early ABEP peaks, generated peripheral to binaural convergence, may be affected by contralateral stimulation. These contralateral effects were in a pattern compatible with suppression. most probably by efferents of the olivo-cochlear bundle. PMID- 9872426 TI - P300 and stress in mild head injury patients. AB - OBJECTIVE: The P300 component of event-related potentials is affected by personal meaningfulness of the stimulus to the subject. Thus, the P300 component could provide an objective parameter in the emotional assessment of road accident mild head injury patients, when exposed to relevant stimuli. METHODS: Thirteen patients with post-traumatic symptoms and 14 healthy controls were evaluated in this study. Two word types, distinguished by color, were presented on a computer screen in active 'oddball' paradigm conditions. In the first subtest, the targets were accident-related (stressful) words; in the second subtest, the targets were non-accident-related (neutral) words. Target (20%) and non-target (80%) were defined by word color. Data recorded from Pz were analyzed for P300 parameters. RESULTS: Patients and controls differed in their reaction to word types (group x word main effect P = 0.0089), regardless of the oddball presentation. Overall, accident-related words produced a significantly larger P300 wave than neutral words in patients (P = 0.0001), but not in controls (P = 0.5741). Significant correlation was found between combined P300 amplitude difference (all stressful words vs. all neutral words) and the patient's Zung state anxiety score (r = 0.68, P = 0.01). CONCLUSION: We suggest the P300 component can provide a useful, objective tool in the assessment of mild head injury patients. PMID- 9872427 TI - Identification of the stimulated hemiretinae using a reduced number of PVEP trials. AB - Left and right hemifield pattern-reversal visual evoked potentials (PVEP) at P100 latency are characterised by typical field distributions of potential and the spectral parameter instantaneous frequency (IF). Both parameters can be utilised for the correct identification of the stimulated hemiretina in healthy volunteers (Hoffmann, K. et al. Electroenceph. clin. Neurophysiol., 100, 1996: 569-578). The aim of this study was the investigation of the robustness of instantaneous frequency for reduced numbers of averages. Hemifield PVEP of 15 volunteers (20 channel records) were analysed. The number of averages was reduced step-by-step (64-32-16-8-4-1). For each average, the time of P100 latency was determined by the Global field power maximum between 90 and 125 ms. The stimulated hemifield was identified using the potential or instantaneous frequency values at the occipital electrode positions O1 and O2: by maximal potential and instantaneous frequency on the stimulus-contralateral side. In summary, by reduced numbers of averages (as well as by single trials) the stimulated hemiretina was correctly identified more frequently on the basis of instantaneous frequency than of potential distribution. A number of 8 averages seems to be sufficient for the correct identification of the stimulus condition. Consequently, for the identification of left and right hemifield stimulations the recording time could be reduced immensely. Instantaneous frequency is suggested as an additional and robust parameter for the selective averaging of artefact-free trials during the recording of hemifield PVEP. PMID- 9872428 TI - Event-related potentials in patients with temporal lobe epilepsy reveal topography specific lateralization in relation to the side of the epileptic focus. AB - Event-related potentials (ERPs) were recorded during a continuous recognition memory paradigm in patients with left-sided (LTLE; n = 8) or right-sided temporal lobe epilepsy (RTLE; n = 6), and in healthy control subjects (n = 24). Control subjects and both patient groups exhibited consistent OLD/NEW ERP-differences from 200-600 ms after stimulus onset. ERPs did not differ significantly between LTLE and RTLE patients, with respect to OLD/NEW distinction or the type of presented material (verbal vs. non-verbal). However, ERP topography showed significant differences between LTLE and RTLE patients: in lateral fronto temporal recordings, patients showed larger negativities contralateral to the seizure focus, whereas we found larger negativities ipsilateral to the seizure focus in parietal recordings. Differences between the groups were significant from 300 to 600 ms post-stimulus. As a consequence, the amplitude gradient from fronto-temporal to parietal recordings was higher on the right side in LTLE patients and on the left side in RTLE patients. Again, differences between LTLE and RTLE patients were highly significant. We assume that ERPs reflect disturbances of a cortico-cortical network dependent on the side of the seizure focus in temporal lobe epilepsy. Furthermore, scalp-recorded ERPs might be a useful tool in the prediction of the side of the seizure focus in patients with temporal lobe epilepsy. PMID- 9872429 TI - Comparison of distortion product otoacoustic emissions with auditory brain-stem response for clinical use in neonatal intensive care unit. AB - OBJECTIVES: This study compares the clinical usefulness of distortion product otoacoustic emissions (DPOAEs) with the auditory brain-stem response (ABR) for neonates in the neonatal intensive care unit for the evaluation of hearing impairment. METHODS: Both DPOAEs and ABR were performed on 36 neonates (67 ears) on the same day. We defined neonates as having normal hearing when the thresholds of wave V of ABR were < or =45 dB hearing level. RESULTS: (1) We could not obtain DPOAEs at f2 = 977 Hz in neonates with normal hearing because of high noise floors. DPOAE recording time was 36 min shorter than that of ABR. (2) We defined as normal DPOAEs, the number of frequencies which showed the DPgram-noise floor > or =4 dB was > or =4 at 6 f2 frequencies, from 1416 Hz to 7959 Hz. (3) Normal thresholds of ABR and normal DPOAEs showed the same percentages, i.e. 68.7%, but the percentage of different results between ABR and DPOAEs was 6.0%. CONCLUSIONS: Our study indicates that DPOAEs represent a simple procedure, which can be easily performed in the NICU to obtain reliable results in high-risk neonates. Results obtained by DPOAEs were comparable to those obtained by the more complex procedure of ABR. PMID- 9872430 TI - P9 in median nerve SEPs is a junctional potential generated by the change of the volume conductor size between trunk and neck. AB - OBJECTIVES: We aimed to investigate the origin of P9 in median SEPs by applying the junctional potential theory. METHODS: We studied the distribution over the body surface with contralateral shoulder reference in 4 normal subjects. RESULTS: A stationary potential field P9/tN9 (=truncal N9) was recorded: P9 over head and neck (the smaller part), tN9 over trunk (the larger part), the boundary being located between trunk and neck. This polarity agreed with that expected from simulation studies. CONCLUSIONS: P9 is a junctional potential generated by the change of the volume conductor size between trunk and neck. PMID- 9872431 TI - Brain electrical source analysis of primary cortical components of the tibial nerve somatosensory evoked potential using regional sources. AB - Tibial nerve somatosensory evoked potentials (SEPs) show higher amplitudes ipsilateral to the side of stimulation, whereas subdural recordings revealed a source in the foot area of the contralateral hemisphere. We now investigated this paradoxical lateralization by performing a brain electrical source analysis in the P40 time window (34-46 ms). The tibial nerve was stimulated behind the ankle (8 subjects). On each side, 2048 stimuli were applied twice. SEPs were recorded using 32 magnetic resonance imaging (MRI)-verified electrode positions (bandpass 0.5-500 Hz). In each case, the P40 amplitude was higher ipsilaterally (0.45 +/- 0.14 microV) than contralaterally (-0.49 +/- 0.16 microV). The best fitting regional source, however, was always located in the contralateral hemisphere with a mean distance of 8.2 +/- 4.3 mm from the midline. The positivity pointed ipsilaterally shifting from a frontal orientation (P37) to a parietal direction (P40). The P40 dipole moment was 2.5 times stronger than the dipole moment of P37, which makes P40 most prominent in EEG recordings. However, with its oblique dipole orientation compared to the tangential P37 dipole, it is systematically underestimated in MEG. Dipole orientations explained interindividual variability of scalp potential distribution. SEP amplitudes were smaller when generated in the dominant (left) hemisphere. This is explained by deeper located sources (5.4 +/- 1.6 mm) with a more tangential orientation (delta theta = 17.5 +/- 2.3 degrees) in the left hemisphere. PMID- 9872432 TI - Acute and chronic neuropathies: new aspects of Guillain-Barre syndrome and chronic inflammatory demyelinating polyneuropathy, an overview and an update. AB - During the last 15 years new information about clinical, electrophysiological, immunological and histopathological features of acute and chronic inflammatory neuropathies have emerged. Thus, the Guillain-Barre syndrome (GBS) is no longer considered a simple entity. Subtypes of the disorder besides the typical predominant motor manifestation, are recognized, i.e. a cranial nerve variant with ophthalmoplegia, ataxia and areflexia, an immune-mediated primary motor axonal neuropathy (AMAN), and a motor-sensory syndrome (AMSAN). Also, the clinical pattern of GBS is related to preceding viral or bacterial infections. Two types of acute motor paralysis have been described, one with slow and incomplete recovery, another with recovery times identical with acute inflammatory demyelinating polyneuropathy (AIDP). Histologically, the first is characterized by Wallerian degeneration of motor roots and peripheral motor nerve fibres. In the latter anti-GM antibodies bind to the nodes of Ranvier producing a failure of impulse transmission. Motor-point biopsies have shown denervated neuromuscular junctions and a reduced number of intramuscular nerve fibres. Molecular mimicry has been postulated as a possible mechanism triggering GBS. Thus, in the cranial variant antibodies to ganglioside GQ1b recognizes similar epitopes on Campylobacter jejuni strains and similar observations apply to anti GM1 antibodies. Chronic inflammatory demyelinating polyneuropathy (CIDP) also has several different clinical presentations such as a pure motor syndrome, a sensory ataxic variant, a mononeuritis multiplex pattern, relapsing GBS, and a paraparetic subtype. Each of the acute and the subtypes have different, more or less distinct, electrophysiologic and pathological findings. Instructive patient stories are presented together with there electrophysiologic and biopsy findings. PMID- 9872433 TI - Autonomic and hormonal ictal changes in gelastic seizures from hypothalamic hamartomas. AB - OBJECTIVES: We describe two patients with hypothalamic hamartoma and gelastic seizures. METHODS: We performed ictal neurophysiological studies with polygraphic recordings of autonomic parameters and hormonal ictal plasma concentration measurements. RESULTS: Ictal recordings showed a stereotyped modification of autonomic parameters: increase in blood pressure and heart rate, peripheral vasoconstriction and modification of respiratory activity. At seizure onset, the norepinephrine plasma level was high and epinephrine unchanged, whereas prolactin and adrenocorticotropic hormone were increased in both cases. Growth hormone and cortisol plasma concentrations in each patient showed a different response to seizures. CONCLUSIONS: These data provide evidence that gelastic seizures are accompanied by an abrupt sympathetic system activation, probably due to the direct paroxysmal activation of limbic and paralimbic structures or other autonomic centres of the hypothalamus and medulla. PMID- 9872434 TI - Categorization of interictal epileptiform potentials using a graph-theoretic method. AB - OBJECTIVES: In patients with epileptic seizures, localization of the source of interictal epileptiform activity is of interest. For correct source localization, a favorable signal to noise ratio is important, and to achieve this, averaging of several epileptiform potentials is often necessary. Before averaging, a careful categorization of epileptiform potentials with different potential distributions is crucial. The aim of this study was to investigate whether a a hierarchic, graph-theoretic algorithm could be used for this categorization. METHODS: In 4 patients, 50-100 sharp waves with different surface distributions were categorized independently with the algorithm, and by visual inspection of the traces. As an independent evaluation of the algorithm, a dipole reconstruction was performed for each sharp wave, and the dipole results for the sharp waves from the different automatically obtained categories were compared. RESULTS: All patients showed a high degree of correspondence between the results of the automatic analysis and the visual estimation. There were clear differences in dipole results between the sharp waves of the different categories obtained from the automatic categorization. CONCLUSION: The results indicate that the graph theoretic categorization algorithm provides a reliable clustering of interictal epileptiform potentials, and that the method may become a useful tool in the pre averaging categorization of interictal epileptiform potentials prior to source localization. PMID- 9872435 TI - Auditory and somatosensory evoked potentials in coma following spontaneous cerebral hemorrhage: early prognosis and outcome. AB - The aim of this study was to check the prognostic power of auditory brain-stem responses (ABRs) and somatosensory evoked potentials (SEPs) in coma following spontaneous cerebral hemorrhage. Seventy patients comatose following subarachnoidal or hypertensive hemorrhage were submitted to ABR and SEP recordings during the acute phase of clinical course. Twenty-one patients survived (30%), two remained vegetative (2.9%) and 47 died (68.1%). The Glasgow Coma Score (GCS) was significantly related to the outcome (P < 0.001), but showed a low sensitivity, since about 50% of patients with GCS = 5-8 died or remained vegetative. ABRs and SEPs showed a much closer correlation with outcome (P < 0.001): their combined use allowed there to be a sensitivity of 96%, a specificity of 90% and a predictive power of 96%; the relative risk of poor outcome in patients with at least one abnormal modality was equal to 223 times the one for patients with normal evoked potentials. Moreover, in surviving patients a significant relationship appeared to exist between abnormalities of SEPs during the acute phase and the severity of disability. Our results confirm the prognostic effectiveness of short latency evoked potentials in cerebral hemorrhage: they are far superior to clinical data, being able to yield a marked decrease of falsely optimistic predictions. PMID- 9872436 TI - EEG changes in intrathecal baclofen overdose: a case report and review of the literature. AB - OBJECTIVE: To review the clinical and EEG manifestations of intrathecal baclofen overdose. METHODS: We identified one patient who had received an overdose of intrathecal baclofen. Information about the clinical course was obtained by reviewing the patient's medical record. EEGs were recorded with the use of the standard 10-20 electrode placement system. RESULTS: The patient received 30 mg baclofen intrathecally. Shortly after the injection he developed respiratory insufficiency and quadriparesis and later became comatose. The first EEG obtained 20 h after the injection showed very frequent quasiperiodic generalized epileptiform discharges. The patient gradually improved clinically and a second EEG obtained 24 h later showed only intermittent bursts of generalized slow wave activity. A repeat EEG study 1 week later was normal. CONCLUSIONS: The EEG in intrathecal baclofen overdose can show quasiperiodic generalized epileptiform discharges. This does not necessarily indicate the presence of underlying potential epileptogenicity, and treatment with an antiepileptic medication is not necessary. PMID- 9872437 TI - EEG source localization: implementing the spatio-temporal decomposition approach. AB - OBJECTIVES: The spatio-temporal decomposition (STD) approach was used to localize the sources of simulated electroencephalograms (EEGs) to gain experience with the approach for analyzing real data. METHODS: The STD approach used is similar to the multiple signal classification method (MUSIC) in that it requires the signal subspace containing the sources of interest to be isolated in the EEG measurement space. It is different from MUSIC in that it allows more general methods of spatio-temporal decomposition to be used that may be better suited to the background EEG. RESULTS: If the EEG data matrix is not corrupted by noise, the STD approach can be used to locate multiple dipole sources of the EEG one at a time without a priori knowledge of the number of active sources in the signal space. In addition, the common-spatial-patterns method of spatio-temporal decomposition is superior to the eigenvector decomposition for localizing activity that is ictal in nature. CONCLUSIONS: The STD approach appears to be able to provide a means of localizing the equivalent dipole sources of realistic brain sources and that, even under difficult noise conditions and only 2 or 3 s of available EEG, the precision of the localization can be as low as a few mm. PMID- 9872438 TI - Electroencephalographic discharges of temporal lobe seizures in children and young adults. AB - We investigated the discharge morphology and propagation patterns of electroencephalographic seizures of temporal lobe onset in 21 children and young adults who underwent invasive long-term EEG monitoring (LTM). Of those, 15 subsequently underwent anterior temporal lobectomy. The onset was focal in 63%. The most frequent discharge morphology was low amplitude beta (30%) or rhythmic/semirhythmic theta discharge (30%). Thirteen patients displayed several sequences of propagation with different spreading stages along a fixed path. Initial spreading to the ipsilateral frontal lobe was associated with a higher frequency of secondary generalization than initial spreading to the contralateral temporal lobe (P = 0.18). A comparison of 13 patients older than 18 years of age with 8 patients younger than 14 years showed a trend towards a lower rate of propagating from the temporal lobe (P = 0.13) in the younger age group. Discharge morphology was not correlated with age, focality, or outcome of surgery. PMID- 9872440 TI - EEG findings during basilar migraine attacks in children. AB - We present clinical and EEG findings in 4 children with basilar migraine (BM) (three female and one male, age 11 to 13.5 years). All patients had an EEG during the acute attack and a follow-up EEG within 4 to 18 days. In two patients the EEG, done within 4 h of the onset of symptoms (initial stage), showed diffuse polymorphic subdelta-delta activity. In two other children the EEG, performed 16 h after the onset of symptoms, showed delta-theta activity predominant over the occipital regions. Resolution of these abnormalities during follow-up was observed in all patients. We wish to stress the danger of misinterpretation of the slow wave activity in the EEG of patients with BM attacks. Together with the clinical findings and their evolution, EEG results should not be interpreted as a sign of a structural brain-stem lesion, such as infarction or inflammation. PMID- 9872439 TI - Physiological bases of the synchronized population spikes and slow wave of the magnetic field generated by a guinea-pig longitudinal CA3 slice preparation. AB - OBJECTIVE: The physiological bases of evoked magnetic fields were examined in a guinea-pig hippocampal slice preparation, motivated by new concepts in central nervous system (CNS) electrophysiology brought about by discoveries of active conductances in the dendrites and soma of neurons. METHODS: Their origins were elucidated by comparing them with intracellular and extracellular field potentials. RESULTS: With excitatory synaptic transmissions blocked, the magnetic signal elicited by an electrical stimulus applied to the pyramidal cell layer consisted of a spike and a depolarizing afterpotential-like waveform. With the excitatory synaptic transmissions intact, but with inhibitory synaptic transmissions blocked, the magnetic signal was bi- or triphasic depending on whether the cell layer or the apical dendrite area of the pyramidal cells was, respectively, depolarized. In both cases the signal consisted of a train of synchronized population spikes superimposed on a brief wave followed by a longer, slow wave. The spike train was correlated with synaptically mediated intracellular spikes. The underlying currents for the slow wave were directed from the apical to the basal side for both types of stimulation. It was most likely generated by depolarization of the apical dendrites, caused by recurrent excitatory synaptic activation. CONCLUSIONS: This analysis illustrates how synaptic connections and intrinsic conductances in a disinhibited mammalian CNS structure can generate spikes and waves of the magnetic field and electrical potential. PMID- 9872441 TI - Neurophysiological evaluation of pain. AB - Neurophysiological techniques for the evaluation of pain in humans have made important advances in the last decade. A number of features of neuroanatomy and physiology of nociception qualifies pain as a multidimensional phenomenon which is rather unique among the sensory systems and which poses a number of technical and procedural requirements for its appropriate diagnostic assessment. Various stimulation techniques to induce defined pain in humans and used in combination with the methodology of evoked electrical brain potentials and magnetic fields are presented. Most recent knowledge gathered from scalp topography and dipole source analysis of pain-relevant evoked potentials and fields is discussed. Particular emphasis is put upon laser-evoked potentials and their application for diagnosis, pathophysiological description and monitoring of patients with neurological disorders and abnormal pain states. Future perspectives in this growing field of research are discussed briefly. PMID- 9872442 TI - Influence of spontaneous activity on peak-ratio analysis. AB - OBJECTIVES: To prove or disprove this assumption that in neuropathy patients with abundant spontaneous activity, peak-ratio interference pattern analysis may lead to false negative results. METHODS: Spontaneous activity >100 microV, automatically analysed by turn/amplitude analysis and expressed as (turns/second)/2 ((T/S)/2), and interference patterns, analysed by the peak-ratio technique, were recorded, one after the other, from the right anterior tibial muscle of 21 patients with neuropathy, aged 36-87 years. RESULTS: The mean number of spontaneous discharges ((T/S)/2) was 12.3 (range 5.5-26) and its mean amplitude 261 microV (range 146-478 microV). Despite this abundant spontaneous activity, peak-ratio analysis was neurogenic in 81% of the patients. All peak ratio parameters were independent on the amount and amplitude of spontaneous discharges. CONCLUSIONS: Spontaneous discharges >100 microV could be adequately assessed by means of the turn/amplitude analysis and did not influence peak-ratio analysis in neuropathies. PMID- 9872443 TI - Characteristic EEG findings in ring 20 syndrome as a diagnostic clue. AB - OBJECTIVE: To review the EEG features of ring 20 syndrome in two patients and determine the characteristic pattern of this syndrome. The features of our cases and 24 patients reported in the literature will be discussed. SUBJECTS AND METHODS: Report of two patients and review of literature. RESULTS: The two patients had intractable epilepsy since childhood. Their clinical seizures were mostly complex partial seizures. Often the patients seizures were of prolonged duration. Ictal EEG revealed characteristic slow waves, and sharp waves. The slow waves were (1) usually synchronous high-voltage slow waves with or without a spike component predominantly in the frontal and frontopolar areas, (2) sometimes showed a change in frequency every several seconds, (3) continued for a long period, and (4) easily spread diffusely. The sharp waves were 5-6 Hz irregular and diffuse discontinuous sharp waves, and sometimes appeared predominantly in the centroparietal area. The clinical seizure pattern and EEG findings were similar in the 24 published cases. CONCLUSIONS: These EEG findings may be a characteristic feature of ring 20 syndrome and thus may be useful as a diagnostic clue. PMID- 9872444 TI - Inter-hemispheric lateralization of event related potentials; motoric versus non motoric cortical activity. AB - To study hemispheric lateralization of cortical potentials associated with motoric and non-motoric function, cortical activity was recorded accompanying either finger extension or saccadic eye movements in a contingent negative variation (CNV) paradigm. Subjects viewed computer-generated pacing stimuli, presented in the left visual hemi-field, and were instructed to either initiate or inhibit a motor response following an imperative signal. Motoric lateralization was assessed by means of the lateralized readiness potential (LRP). In addition, a measure complementary to the LRP was introduced to investigate non-motoric lateralization (NML). Contralateral inter-hemispheric lateralization was evident in the LRP preceding finger movement, but was absent prior to eye movements. However, pre-saccadic cortical response profiles did exhibit a right hemispheric, non-motoric lateralization (NML) during stimulus presentation. Comparable non-motoric lateralization was found for finger extension. Results of the present study suggest that non-motoric lateralization may be a contributing factor to the frequently reported inter-hemispheric asymmetry preceding self-initiated saccadic eye movements. Results of the present study also suggest that the latter may be related to a covert shift of visuospatial attention toward the saccadic target. Associated shifts of attention are suppressed in a CNV paradigm, where attentional focus is primarily on the CNV stimulus during the pre-saccade period. PMID- 9872445 TI - Intracerebral recording of potentials accompanying simple limb movements: a SEEG study in epileptic patients. AB - OBJECTIVES: Slow potentials appearing during simple repetitive acral limb movement were investigated. Twenty-six patients suffering from drug resistant partial epilepsies and explored with implanted intracerebral electrodes were examined using two protocols. METHODS: In 18 patients, readiness potential (RP), in 13 patients contingent negative variation (CNV), and in 7 patients both protocols, were tested. The recordings from leads with evident pathological EEG activity were excluded from evaluation. The results concerning the slow potentials preceding the movements in RP and CNV protocols have already been published. RESULTS: The movement-accompanying slow potentials (MASP) were polyphasic or monophasic, started before or during the movement. In the primary motor cortex they followed the pre-movement potentials depending on the protocol: in the RP paradigm they were present only contralateral to the movement, but were bilateral in the CNV protocol. In other areas they either followed the potentials preceding the movement, in some cases with opposite polarity, or they occurred alone. MASP was recorded in motor and supplementary motor, premotor and prefrontal, midtemporal, somatosensory, superior parietal and cingular cortices. The cingular cortex was heavily involved in the self-paced movements but rarely in the cued movements. CONCLUSION: The major involvement of the cingular gyrus contrasted with the absence of slow potentials in temporal limbic structures. MASP is evidently a heterogenic phenomenon. Its genesis could be involved in a spread of information through the relevant structures. PMID- 9872446 TI - Perception of motion and qEEG activity in human adults. AB - This study was designed to relate visual perception of motion to cortical activity, by evaluation of the association of quantified electroencephalogram (qEEG) parameters with a video film projection. The EEG was recorded from 14 sites according to the International 10-20 system and a common average reference was used. Forty right-handed volunteers (mean age = 24 years) were examined. The video film consisted of 20 s sequences showing still shots and moving shots with human movements or object movements. The EEG was then subjected to spectral analysis; the spectral powers for the theta, alpha and beta bands were calculated for 14 s epochs and compared with sequences of the video film. All analyses were based on logarithmically transformed absolute spectral power values. The power values of each frequency band were analysed in a 3-way repeated measure ANOVA (Hemisphere x Electrode x Sequence). The results were represented by EEG cartography. Significant decreases in the alpha 1, beta 1 and beta 2 power values of EEG in centro-parietal regions of both hemispheres were shown during perception of human motion sequences. This suggests participation of the sensorimotor cortex during visual observation of human motion. PMID- 9872447 TI - Is EEG useful in assessing patients with acute encephalitis treated with acyclovir? AB - EEG has been used widely in diagnosing encephalitis, as it demonstrates rather typical abnormalities, especially in herpes simplex virus encephalitis (HSVE). We analysed 204 EEG recordings from 98 consecutive acyclovir-treated patients with acute encephalitis between 1984 and 1994. Periodic complexes (PC) in the acute phase predicted poor outcome (Kendall tau 0.40, P < 0.001). However, unlike in many other diseases, e.g. stroke and intracerebral haemorrhage, the diffuse slowing of the background activity at acute phase did not predict outcome (Kendall tau -0.6, P = 0.35). At follow-up, the emergence of diffuse slow background activity was significantly associated with a less favourable outcome (Kendall tau 0.33, P = 0.0016). Among clinical variables, only epileptic seizures early during the course of the disease correlated significantly with outcome. EEG does have value as a prognostic indicator in acute encephalitides, but it seems that diffuse slowing of background activity or irritative features acutely are not as important as previously thought, based on the experiences of the pre acyclovir era. PMID- 9872448 TI - 35th annual meeting of the SENFC (Spanish Society of Clinical Neurophysiology). December 11-13, 1997, Barcelona, Spain. PMID- 9872449 TI - Genes from nine genomes are separated into their organisms in the dinucleotide composition space. AB - A set of 16 kinds of dinucleotide compositions was used to analyze the protein encoding nucleotide sequences in nine complete genomes: Escherichia coli, Haemophilus influenzae, Helicobacter pylori, Mycoplasma genitalium, Mycoplasma pneumoniae, Synechocystis sp., Methanococcus jannaschii, Archaeoglobus fulgidus, and Saccharomyces cerevisiae. The dinucleotide composition was significantly different between the organisms. The distribution of genes from an organism was clustered around its center in the dinucleotide composition space. The genes from closely related organisms such as Gram-negative bacteria, mycoplasma species and eukaryotes showed some overlap in the space. The genes from nine complete genomes together with those from human were discriminated into respective clusters with 80% accuracy using the dinucleotide composition alone. The composition data estimated from a whole genome was close to that obtained from genes, indicating that the characteristic feature of dinucleotides holds not only for protein coding regions but also noncoding regions. When a dendrogram was constructed from the disposition of the clusters in the dinucleotide space, it resembled the real phylogenetic tree. Thus, the distinct feature observed in the dinucleotide composition may reflect the phylogenetic relationship of organisms. PMID- 9872450 TI - Phosphate transporter gene family of Arabidopsis thaliana. AB - Using a high-affinity phosphate transporter gene of Arabidopsis thaliana, PHT1, as a probe, three Arabidopsis homologs were isolated, named PHT4, PHT5 and PHT6, in addition to the previously isolated PHT2 and PHT3. The amino acid sequences deduced from the three nucleotides were 32-42% homologous with microbial phosphate transporters of Saccharomyces cerevisiae (PHO84), Neurospora crassa (PHO-5) and Glomus versiforme (GvPT). PHT1, PHT2, PHT3 and PHT6 were clustered in a 25-kbp region of chromosome V. PHT1 and PHT4 transcripts were detected in roots. Interestingly, suspension-cultured cells expressed only PHT4. PHT4 and PHT5 located within a genetic distance of 6.4 cM on chromosome II, and were close to a phosphate accumulation mutant pho2. Genomic sequencing revealed no difference in the sequences of the two genes in both pho2 and wild-type. The PHT4 transcript was expressed at similar levels in the mutant and wild-type. These results demonstrate that neither PHT4 nor PHT5 is allelic to PHO2. PMID- 9872451 TI - A chromosome 4 satellite I DNA isolated from SV40-transformed human cells. AB - Analysis of cellular DNA insert isolated from a free replicative plasmid rescued from human cells transformed with an SV40 vector plasmid revealed the presence of two arrays of repetitive DNA arranged in tandem. One sequence was homologous to the consensus sequence of the human alpha satellite DNA and the adjoining sequence was a satellite DNA sequence which consisted of repetitive units of 42 base pairs (bp) and was designated HR42. The degree of homology between repetitive units was about 92%. By Southern analysis the HR42 sequence was detected in HHW416, a somatic cell hybrid containing human chromosome 4, but not in HDm-5, the somatic cell hybrid which has human chromosome 14. By fluorescence in situ hybridization this repetitive DNA was assigned uniquely to the centromeric region of human chromosome 4. These results show that HR42 belongs to a subfamily of satellite I DNA specific for human chromosome 4. PMID- 9872452 TI - Prediction of the coding sequences of unidentified human genes. XI. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. AB - In our series of projects for accumulating sequence information on the coding sequences of unidentified human genes, we have newly determined the sequences of 100 cDNA clones from a set of size-fractionated human brain cDNA libraries, and predicted the coding sequences of the corresponding genes, named KIAA0711 to KIAA0810. These cDNA clones were selected according to their coding potentials of large proteins (50 kDa and more) in vitro. The average sizes of the inserts and corresponding open reading frames were 4.3 kb and 2.6 kb (869 amino acid residues), respectively. Sequence analyses against the public databases indicated that the predicted coding sequences of 78 genes were similar to those of known genes, 64% of which (50 genes) were categorized as proteins functionally related to cell signaling/communication, cell structure/motility and nucleic acid management. As additional information concerning genes characterized in this study, the chromosomal locations of the clones were determined by using human rodent hybrid panels and the expression profiles among 10 human tissues were examined by reverse transcription-coupled polymerase chain reaction which was substantially improved by enzyme-linked immunosorbent assay. PMID- 9872453 TI - Quick identification and localization of CpG islands in large genomic fragments by partial digestion with Hpa II and Hha I. AB - More than 50% of mammalian genes are associated with CpG islands and thus they serve as a good gene marker. We have devised a simple method to scan large pieces of native or cloned genomic DNA for CpG islands. The method is based on the presence of multiple Hpa II and Hha I sites in CpG islands, at a frequency 30 times higher than in the rest of the genome. The steps include complete digestion of DNA with a rare-cutting restriction endonuclease (to produce large fragments with defined ends), partial digestion with Hpa II and Hha I, and subsequent Southern hybridization with an end probe. This identifies a CpG island as a cluster of sub-bands and, based on their electrophoretic mobility, one can immediately locate the island relative to the ends. For many vectors, universal probes flanking the cloning site are available, enabling the simultaneous analysis of a large number of samples. We demonstrated the usefulness of the method by analyzing known CpG islands in native genomic DNA and lambda, cosmid and P1 clones, and by isolating two novel transcribed islands from anonymous cosmid clones. Our method is quick, inexpensive, and can detect CpG islands with few or even no rare-cutter sites. PMID- 9872454 TI - Structural analysis of Arabidopsis thaliana chromosome 5. VII. Sequence features of the regions of 1,013,767 bp covered by sixteen physically assigned P1 and TAC clones. AB - Sixteen P1 and TAC clones assigned to Arabidopsis thaliana chromosome 5 were sequenced, and their sequence features were analyzed using various computer programs. The total length of the sequences determined was 1,013,767 bp. Together with the nucleotide sequences of 109 clones previously reported, the regions of chromosome 5 sequenced so far now total 9,072,622 bp, which presumably covers approximately one-third of the chromosome. A similarity search against the reported gene sequences predicted the presence of a total of 225 protein-coding genes and/or gene segments in the newly sequenced regions, indicating an average gene density of one gene per 4.5 kb. Introns were identified in 72.4% of the potential protein genes for which the entire gene structure was predicted, and the average number per gene and the average length of the introns were 3.3 and 163 bp, respectively. These sequence features are essentially identical to those in the previously reported sequences. The sequence data and gene information are available on the World Wide Web database KAOS (Kazusa Arabidopsis data Opening Site) at http://www.kazusa.or.jp/arabi/. PMID- 9872455 TI - cDNA cloning, tissue distribution and chromosomal localization of the human ID4 gene. AB - A cDNA e encoding the human Id4 protein has been isolated from an astrocytoma library. The predicted protein product shares 98% identity with the mouse Id4 protein and is markedly different from that already reported. By FISH analysis, the human ID4 gene was more precisely mapped to chromosome 6p22.3-p23. Northern blot analysis showed that ID4 is mainly expressed in thyroid, brain and fetal tissue and in some nervous system tumor cell lines. PMID- 9872456 TI - Cloning of a novel rat gene, DB83, that encodes a putative membrane protein. AB - Using a partial cDNA sequence and a 5'-RACE technique, we isolated a novel cDNA from rat liver referred to as DB83. DB83 had four hydrophobic trans-membrane domains and one N-myristoylation site as well as multiple possible phosphorylation sites. The db83 gene was highly expressed in the liver and significantly in brain, lungs and kidneys. We suggest that DB83 is a tissue specific putative membrane protein. p6 PMID- 9872458 TI - Alterations in temporal/spatial distribution of GFAP- and vimentin-positive astrocytes after spinal cord contusion with the New York University spinal cord injury device. AB - Astrocytes become reactive as a result of various types of lesions and upregulate 2 intermediate filaments, glial fibrillary acidic protein (GFAP), and the developmentally regulated protein vimentin. Young female Sprague-Dawley rats were subjected to a spinal cord contusion at segment T10 using the New York University injury device. Animals were killed at 1, 2, 7, 14, and 30 days postinjury. Horizontal spinal cord sections spanning segments T7-T13 were assessed with antibodies to both intermediate filament proteins. The number of gray matter GFAP positive astrocytes increased by 2 days postinjury, with segments adjacent (proximal) to the injury site showing greater responses than areas several segments away (distal). By 30 days following injury, astroglial cell numbers returned to normal levels. Vimentin-positive astrocytes also showed a graded proximal/distal response by 2 days following injury. Proximal regions remained significantly higher at 30 days following injury than control animals. Rostral/caudal changes were also evident, with regions caudal to the injury showing significantly higher numbers of vimentin positive astrocytes than those rostral, indicating that gray matter areas caudal to spinal cord injury may undergo more stress following spinal cord injury. PMID- 9872457 TI - Role of cyclooxygenase 2 in acute spinal cord injury. AB - Cyclooxygenase, or prostaglandin G/H synthase, is the rate-limiting step in the production of prostaglandins. A new isoform, cyclooxygenase-2 (COX-2), has been cloned that is induced during inflammation in leukocytes and by synaptic activity in neurons. The objectives of this study are to determine the nature of COX-2 expression in normal and traumatized rat spinal cord, and to determine the effects of selective COX-2 inhibition on functional recovery following spinal cord injury. Using a weight-drop model of spinal cord injury, COX-2 mRNA expression was studied with in situ hybridization. COX-2 protein expression was examined by immunohistochemistry and Western analysis. Finally, using the highly selective COX-2 inhibitor, 1-[(4-methylsufonyl)phenyl]-3-tri-fluro-methyl-5-[(4 flur o)phenyl]prazole (SC58125), the effect of COX-2 inhibition on functional outcome following a spinal cord injury was determined. COX-2 was expressed in the normal adult rat spinal cord. COX-2 mRNA and protein production were increased following injury with increases in COX-2 mRNA production detectable at 2 h following injury. Increased levels of COX-2 protein were detectable for at least 48 h following traumatic spinal cord injury. Selective inhibition of COX-2 activity with SC58125 resulted in improved mean Basso, Beattie, and Bresnahan scores in animals with 12.5- and 25-g/cm spinal cord injuries; however, the effect was significant only for the 12.5g/cm injury group (p=0.0001 vs. p=0.0643 in the 25-g/cm group). These data demonstrate that COX-2 mRNA and protein expression are induced by spinal cord injury, and that selective inhibition of COX-2 improves functional outcome following experimental spinal cord injury. PMID- 9872459 TI - Effect of spinal cord transection on N-methyl-D-aspartate receptors in the cord. AB - Spinal cord injury can lead to an exaggeration of transmission through spinal pathways, resulting in muscle spasticity, chronic pain, and abnormal control of blood pressure and bladder function. These conditions are mediated, in part, by N methyl-D-aspartate (NMDA) receptors on spinal neurons, but the effects of cord injury on the expression or function of these receptors is unknown. Therefore, antibodies to the NMDA-R1 receptor subunit and binding of [3H]MK-801 were used to assess NMDA receptors in the spinal cord. Receptor density in rats with intact spinal cords was compared to that in rats 1 and 2 weeks after spinal cord transection (SCT) at the mid-thoracic level. At 1 and 2 weeks after SCT, [3H]MK 801 binding was reduced in most laminae in cord segments caudal to the injury, whereas no decrease in amount of R1 subunit immunoreactivity was observed. No significant changes in [3H]MK-801 binding and NMDA-R1 immunoreactivity could be seen rostral to the transection. Since [3H]MK-801 binding requires an open ion channel, the discrepancy between [3H]MK-801 binding and immunocytochemistry may indicate a loss of functional receptors without a consistent change in their total number. Therefore, the exaggerated reflexes that are well established in rats 2 weeks after cord injury must be mediated by a mechanism that withstands attenuation of NMDA receptor function. PMID- 9872460 TI - Effect of traumatic brain injury on mouse spatial and nonspatial learning in the Barnes circular maze. AB - Controlled cortical impact (CCI) is a relatively new model of traumatic brain injury in the mouse, which, in combination with behavioral and histological methods, has potential for elucidating underlying mechanisms of neurodegeneration using genetically altered animals. Previously, we have demonstrated impaired spatial learning in a water maze task following CCI injury at a moderate level. There are many difficulties associated with this task, however, such as stress, physical demand, and the multiple trials over days required for satisfactory training. As a potential alternative to the water maze, we adapted the Barnes circular maze to our mouse model and assessed spatial/nonspatial learning following injury. Mice were trained to locate a dark tunnel, hidden beneath one of 40 holes positioned around the perimeter of a large, flat, plastic disk, brightly illuminated by four overhead halogen lamps. Sham-operated animals rapidly acquired this task, exhibiting reduced latency to find the tunnel and a more efficient search strategy as compared with injured mice. This difference was not due to visuomotor deficits, as all mice performed equally well in a cued version of the same task. These results demonstrate spatial learning impairment following CCI injury in a task that offers an efficient alternative to the water maze. PMID- 9872462 TI - Effect of glucose administration on contusion volume after moderate cortical impact injury in rats. AB - Previous studies had shown that pre- and postinjury glucose administration increased brain injury caused by a mild cortical impact injury only when the traumatic injury was complicated by a secondary ischemic insult. The purpose of this study was to examine the effect of pre- and postinjury glucose administration on a more severe cortical impact injury, where primary ischemia occurs at the site of the impact. Long Evans rats who were fasted overnight and anesthetized with isoflurane were subjected to a 5-m/sec, 2.5-mm impact injury. The animals were randomly assigned one of the following treatments: (1) 2.2 g/kg glucose in 4 ml of saline, 20 min prior to injury; (2) 2.0 g/kg glucose in 4 ml of saline, 20 min after injury; or (3) 4 ml of saline either 20 min before injury or 20 min after the injury. At 2 weeks, the animals were sacrificed and the brains were examined for contusion volume and for neuronal loss in CA1 and CA3 regions of the hippocampus. Contusion volume was increased from a median value of 23 mm3 in the saline-infused animals to 34 mm3 in the preimpact glucose infusion animals (p=0.005). Postimpact glucose infusion had no effect on contusion volume. Neuron density in CA1 and CA3 regions of the hippocampus was similar in all three treatment groups. These studies support the hypothesis that glucose administration adversely affects experimental traumatic brain injury in those circumstances where the trauma is complicated by primary cerebral ischemia, such as around cortical contusions. PMID- 9872461 TI - The neuronal cytoskeleton is at risk after mild and moderate brain injury. AB - Recent studies have described alterations in cytoskeletal proteins such as microtubule-associated protein 2 (MAP-2) and neurofilament (NF) resulting from moderate and severe experimental brain injury; however, few have investigated the consequences of mild injury, which is associated clinically and experimentally with cognitive dysfunction and neuronal damage. To contrast cytoskeletal changes within 7 days following mild injury with those following moderate injury, we subjected anesthetized, adult rats to mild (1.1-1.3 atm) or moderate (2.3-2.5 atm) lateral fluid percussion brain injury or sham injury. Rats were sacrificed at 6 h (n=4 mild; n=4 moderate; n=2 sham), 24 h (n=4 mild; n=4 moderate; n=1 sham), or 7 days (n=5 mild; n=4 moderate; n=1 sham) following injury, and immunohistochemistry was performed for MAP-2 and NF. Both mild and moderate injury produced notable cytoskeletal changes in multiple brain regions; however, mild injury generally resulted in a lesser degree of MAP-2 and NF loss over a smaller spatial extent. When compared to moderately injured animals, animals subjected to mild injury showed substantially delayed MAP-2 and NF alterations within the cortex and hippocampal dentate gyrus and no evidence of MAP-2 loss in the hippocampal CA3 region. While mild and moderate injury resulted for the most part in similar patterns of axonal injury, tissue tears in the fimbria and loss of NF immunoreactivity in regions containing injured axons were only observed following moderate injury. Elucidating the effects of modulating injury severity may yield insight into the mechanisms involved in traumatic damage to the cytoskeleton and guide future treatment strategies. PMID- 9872463 TI - Reduced reactivity of the middle cerebral artery and its large branches after cold lesion. AB - The aim of this study was to measure vascular reactivity in the isolated middle cerebral artery (MCA) after brain injury. Segments of MCA were prepared from control, sham-operated, and cold-lesioned rats. Cold lesion was induced by application of a precooled (-78 degrees C) copper cylinder (diameter 5 mm) for 60 sec to the intact dura over the parietal cortex. Endothelin-1 (ET-1) (10(-12) to 3 x 10(-7) M) induced a dose-dependent contraction with a pD2 (-log10 EC50) of 8.36+/-0.12 (mean+/-SEM) and an Emax (maximal response) of 2.41+/-0.15 mN (millinewton) at 10(-7) M in sham-operated animals under resting conditions. This maximum contraction induced by 10(-7) M ET-1 was significantly (p < 0.05) reduced 24 and 48 h after cold lesion by 41% and 30%, respectively. After precontraction with 10(-5) M prostaglandin (PG) F2alpha, ET-3 (10(-12) to 10(-8) M) relaxed the MCA with an Emax of 0.42+/-0.07 mN at 10(-8) M and a pD2 of 9.20+/-0.19 in sham operated animals. This relaxation was reduced 24 and 48 h after cold lesion by 19% and 62% at 10(-8) M, respectively. Concentration-effect curves for bradykinin (BK, 10(-8) to 10(-5) M) in uridine triphosphate (UTP, 10(-4) M)-precontracted MCA segments revealed relaxation with a pD2 of 7.08+/-0.10 and an Emax of 0.65+/ 0.06 mN at 10(-6) M in sham-treated animals. This effect of BK was reduced by 35% and 20% at 10(-6) M 24 and 48 h after cold lesion, respectively. In addition, the contractile responses to 124 mM K+, 10(-5) M PGF2alpha and the dilation induced by 10(-3) sodium nitroprusside (SNP) were reduced in MCA segments taken 24 and 48 h after lesion compared with shams. We conclude that attenuation of ET effects can be explained, at least in part, by tachyphylaxis to ETs. The unspecific reduction of vascular reactivity may result from spreading depression. PMID- 9872465 TI - Anterior tibialis tendon ruptures: an outcome analysis of operative versus nonoperative treatment. AB - Ruptures of the anterior tibialis tendon are a rare clinical entity. Case reports in the literature reveal a total of 28 cases. Unfortunately, because of the limited discourse in the orthopaedic literature, there are few guidelines regarding the treatment for these injuries. This study analyzes the treatment of 16 anterior tibialis tendon ruptures. Eight patients in this group had operative treatment of their ruptures, and eight patients had nonsurgical treatment of their ruptures. The average follow-up for the operative and nonoperative patients were 6.68 years and 3.86 years, respectively. The Foot and Ankle Outcome questionnaire provided by the American Academy of Orthopaedic Surgeons and an outcome-based foot score described by Kitaoka et al. were used as the methods of analysis. Our outcome results show no statistically significant difference between operative and nonoperative treatment in anterior tibialis tendon ruptures. The lack of statistical difference between operative and nonoperative groups may be a reflection of the age bimodality present in this study. Elderly low demand patients were treated nonsurgically and young active patients were treated operatively. Therefore, despite a lack of statistical difference present in the outcome of both groups, we still maintain the need to repair/reconstruct anterior tibialis tendon ruptures in young active patients with high functional demands. The deficits present in the nonoperative group, we believe, would not be well tolerated in a young high functional demand patient. Nonsurgical management is an appropriate alternative in low demand elderly patients. PMID- 9872464 TI - Topical axonal transport blocker vincristine prevents nerve injury-induced spinal neuron sensitization in rats. AB - The effect of vincristine (Vin, a fast axonal transport blocker) to prevent any alteration in the excitability of dorsal horn neurons, following peripheral nerve injury, was investigated on 31 rats: 20 with chronic constriction injury (CCI) of the sciatic nerve and 11 sham preparations. In 15 of the 20 CCI rats, a small piece of gelfoam soaked with Vin was applied to the sciatic nerve before ligation (Vin+); in the remaining 5 rats the nerve was ligated without Vin (Vin-). The 11 sham rats were 7 Vin+ and 4 Vin-. The dorsal horn neuronal activity was recorded after 2-3 postoperative (PO) weeks. In the CCI Vin- rats, the neurons showed increased spontaneous activity and hyperresponsiveness to noxious stimulus with prolonged afterdischarges, events considered to signal central neuron sensitization. In the CCI Vin+ rats, the neuronal spontaneous and stimulated activity values were significantly lower (p < 0.001) than in the CCI Vin- rats being comparable to normal values. In sham Vin+ and Vin- rats, the neuronal activities had normal values. Given the crucial role attributed to central neuron sensitization for the development of neuropathic pain, the possibility that vincristine, by blocking the axonal transport, exerts a preventive action on this syndrome is discussed. PMID- 9872466 TI - Outcome study of subjects with insertional plantar fasciitis. PMID- 9872467 TI - Biomechanical consequences of plantar fascial release or rupture during gait: part I--disruptions in longitudinal arch conformation. AB - To examine whether conformational changes induced by plantar fascial division may progress during gait, we loaded the feet of seven cadavers using an apparatus that simulates the actions of the extrinsic plantarflexors. We measured the effects of plantar fasciotomy at two instants in the terminal-stance phase of gait. Radiographic measurements of height of the arch, base length of the arch, and talo first-metatarsal angle were used to assess contributions to arch support made by the plantar fascia, tibialis posterior, peroneus longus and brevis, and digital flexor muscles. Complete fasciotomy caused significant collapse of the arch in the sagittal plane. Early in terminal stance, at the instant after heel off, mean height of the arch decreased from 47 to 45 mm. Late in terminal stance, at the instant preceding contralateral heel strike, mean height of the arch decreased from 46 to 43. Effects of division of the central band, though significant, were mild. Medial base length of the arch increased from 163 to 167 mm in the absence of tibialis posterior contraction at late terminal stance. Arch supporting abilities of the other extrinsic muscles were insignificant. PMID- 9872468 TI - Dislocation of posterior tibial tendon. AB - Dislocation of the posterior tibial tendon has been reported rarely in the English literature. We report a case of traumatic dislocation of the posterior tibial tendon. The diagnosis was delayed for months and was ultimately made by magnetic resonance imaging, which accurately determined the location of the tendon. At the time of surgical exploration, no predisposing anatomic abnormalities were noted. A new pseudoretinaculum had developed that prevented recurrent subluxation or dislocation. Surgical relocation of the tendon and repair of the retinaculum resulted in a good outcome. PMID- 9872469 TI - Optimal computed tomography imaging of the midfoot: an improved technique. AB - A new protocol for computed tomography (CT) imaging of the midfoot is described. This imaging technique places the CT cuts parallel to and perpendicular to the talus-first metatarsal axis, as viewed on the lateral CT scout image. For imaging the midfoot, this technique is an improvement over previously described hindfoot or midfoot protocols. PMID- 9872470 TI - Operative treatment for hallux valgus in children with cerebral palsy. AB - All patients with cerebral palsy who had hallux valgus and bunion deformities surgically corrected between 1986 and 1995 were reviewed to determine whether techniques other than arthrodesis of the first metatarsophalangeal joint would adequately correct these deformities. Twenty-six feet of 17 patients were surgically corrected for their bunions and/or their hallux valgus deformity. The mean hallux valgus angle for all children was 30 degrees, preoperatively, with 70% correction achieved. Four techniques of surgical correction were used. A first metatarsophalangeal joint arthrodesis on eight feet gave excellent outcomes for all using the duPont Bunion Rating Score and a mean of 89% correction of the hallux valgus angle. A proximal first metatarsal osteotomy, distal soft tissue release, and exostectomy of the bunion on five feet gave three excellent and two fair outcomes, with a mean of 83% correction of the hallux valgus angle. A distal soft tissue release and exostectomy on eight feet gave four excellent outcomes, two good outcomes, and two fair outcomes, with a mean of 53% correction of the hallux valgus angle. An osteotomy of the first proximal phalanx, metatarosphalageal soft tissue release, and exostectomy on five feet gave three excellent outcomes, one good outcome, and one fair outcome with a mean of 36% correction of the hallux valgus angle. All patients who met the criteria for the procedures were satisfied with the outcomes; however, the first metatarosphalageal joint arthrodesis gave the best results with the highest percent correction and bunion score. PMID- 9872471 TI - Evaluation of multidensity orthotic materials used in footwear for patients with diabetes. AB - Selected combinations of multidensity orthotic materials were tested under simulated walking conditions found in the forefoot of diabetic patients. Materials were compared for therapeutic effectiveness by their stress/strain properties and dynamic compression set. Results showed that all of the multidensity materials experienced losses in performance throughout the testing period of 100,000 cycles, with the greatest losses occurring within the first 10,000 cycles. Of the materials tested, Poron + Plastazote #2 and Spenco + Microcel Puff Lite had the highest dynamic material strain and the lowest dynamic compression set over 100,000 cycles. In comparison, these are better multidensity combinations than the others tested to use as therapeutic orthoses in footwear for diabetic patients. PMID- 9872472 TI - Intra-articular calcaneal fractures: effect of open reduction and internal fixation on the contact characteristics of the subtalar joint. AB - Intra-articular calcaneal fractures are associated with significant long-term morbidity, and considerable controversy exists regarding the optimum method of treating them. The contact characteristics in the intact subtalar joint were determined at known loads and for different positions of the ankle and subtalar joint, using pressure-sensitive film (Super Low; Fuji, Itochu Canada Ltd, Montreal, Quebec). We measured the contact area to joint area ratio (pressure > 5 kg force/cm2 [kgf/cm2]) which normalizes for differences in joint size and the ratio of high pressure zone (>20 kgf/cm2) as a reflection of overall increase in joint pressure. Three simulated fracture patterns were then created and stabilized with either 1 or 2 mm of articular incongruity. Eight specimens were prepared with a primary fracture line through the posterior facet, eight with a joint depression-type fracture, and six with a central joint depression fracture. A measure of 1 to 2 mm of incongruity in the posterior facet for all three fracture patterns produced significant unloading of the depressed fragment, with a redistribution of the overall pattern of pressure distribution to parts of the facet that were previously unloaded. PMID- 9872473 TI - Total calcanectomy for the treatment of chronic calcaneal osteomyelitis. AB - The purpose of this study is to present the surgical and functional results of a total calcanectomy procedure as a foot salvage alternative in patients with extensive chronic osteomyelitis of the calcaneus. A retrospective review identified eight patients treated with a total calcanectomy for a chronic nonhealing plantar ulcer of the heel and osteomyelitis of the calcaneus. The primary diagnosis was insulin-dependent diabetes mellitus (six patients), pneumococcal septicemia (one patient), and an open calcaneal fracture (one patient). The average age of the patients was 52 years. Prior procedures included irrigation and debridement of the heel ulcer (seven patients), partial calcanectomy (three patients), and split thickness skin grafting (two patients). The vascular status of each limb was assessed preoperatively. Distal extremity bypass surgery was performed on two patients before calcanectomy. Osteomyelitis of the calcaneus was diagnosed by magnetic resonance imaging alone in three patients, and by technetium/indium scans and magnetic resonance imaging in five patients. The average follow-up duration was 27.3 months (range, 6-57 months). Infection at the plantar heel was controlled in all patients. In all eight cases the incisions were closed primarily. During a prolonged time of total contact casting to facilitate wound healing, one patient developed an anterior tibial ulcer that progressed to osteomyelitis and underwent below-knee amputation. Talonavicular subluxation occurred as a late complication in one patient. This was treated with a talonavicular arthrodesis and subsequent bracing for a nonunion. An assessment of functional ambulation was performed on all eight patients. Four patients maintained the same ambulation level postoperatively in a modified heel-containment orthosis. Two patients decreased one functional ambulation level, and one patient decreased two levels. One patient underwent below-knee amputation and is currently ambulatory with a prothesis. Assessment of ankle strength and range of motion of the surgical limb demonstrated decreased dorsiflexion and plantarflexion strength and a variable range of motion compared to the contralateral limb. Total calcanectomy is an alternative procedure to transtibial amputation in patients with chronic osteomyelitis of the calcaneus. Eradication of infection and preservation of the functional ambulation is achieved. PMID- 9872474 TI - Wound-healing risk factors after open reduction and internal fixation of calcaneal fractures. AB - This retrospective study investigated outcomes of wound healing in a series of 63 consecutive patients with 64 fractures of the calcaneus who underwent open reduction and internal fixation done by two surgeons experienced in this fracture during a 3-year period. Thirty-nine patients were managed preoperatively as outpatient referrals before surgery. Twenty-four patients were admitted directly to the trauma service and were managed as inpatients preoperatively. Minimum patient follow-up was 6 months, with an average follow-up of 18 months. A trend correlating the time between injury and operative intervention with the incidence of complications in wounds was noted; the incidence rose in patients who underwent surgery >5 days after their injury. Two-layered closures had a lower incidence of dehiscence compared to single-layered tension-relieving sutures. Patients with a higher body-mass index (BMI) (kg/ m2) took longer to heal their wounds. Strong trends were noted to link BMI and severity of fractures. In the outpatient group, a history of active smoking preoperatively correlated with increased time to wound healing. In 43 patients, there were no wound-healing complications. In 21 feet, there were varying degrees of wound dehiscence. Average wound healing took 47 days. Risk factors for complications in the wound after calcaneal open reduction and internal fixation include single layered closure, high BMI, extended time between injury and surgery, and smoking. Age, type of immobilization, medical illness (including diabetes), type of bone graft, or use of a Hemovac did not influence wound healing. PMID- 9872475 TI - Immunoglobulin variable region sequences of human monoclonal anti-DNA antibodies. AB - OBJECTIVE: Anti-DNA antibodies are believed to be important in the pathogenesis of systemic lupus erythematosus (SLE). Antibodies that bind specifically and with high affinity to dsDNA are most closely involved in tissue damage. Analysis of the sequences of the variable regions of human monoclonal anti-DNA antibodies is useful in defining the structural features that give rise to these binding properties. This article systematically reviews the evidence derived from such sequences. METHOD: Previous reviews of this subject have been hampered by incomplete knowledge of the human immunoglobulin variable region repertoire. In this article, the original sequence data from reports of over 50 human monoclonal antibodies (mAb) are reinterpreted by alignment to the most similar alleles of the most similar germline genes. This allows accurate estimation of the site and nature of somatic mutations. RESULTS: Human IgG monoclonal anti-DNA antibodies generally carry more mutations than IgM. In many cases these have been selected by an antigen-driven process. In many of the more specific, higher affinity dsDNA binders, there is an accumulation of basic residues in the complementarity determining regions. However, many exceptions to this rule exist, particularly among IgM mAb. CONCLUSIONS: Unlike murine anti-DNA antibodies, these human mAb show little evidence for preferential use of particular V(H), V(K) and V(lambda) genes or families to encode antibodies of this specificity. PMID- 9872476 TI - The regulation of heat shock proteins and their role in systemic lupus erythematosus. AB - OBJECTIVES: After a serendipitous suggestion, it was established that a significant subset of patients with systemic lupus erythematosus (SLE) overexpress the 90-kD heat shock protein (Hsp90). In this review, we have analyzed our own data and that of others, to explore the link between Hsp90 and SLE. METHODS: We performed a detailed literature review focusing on the potential role of Hsp in the etiopathogenesis of SLE. RESULTS: Data are discussed showing surface expression of this Hsp in patients with lupus, a similar overexpression in the splenocytes of MRL/Ipr mice before the onset of disease, the detection of antibodies to Hsp90 in a proportion of both lupus patients and lupus-prone mice, and most recently, an analysis of the transcription factors that regulate the production of this protein and the influence of key cytokines on these factors. CONCLUSIONS: These observations provide a model to show how a protein with key physiological roles in healthy individuals may, on occasion, become the target of an autoimmune attack with clinical consequences recognized in both mouse and human. Given that up to now, other heat shock proteins are not targeted in a similar fashion, the specificity of these responses is remarkable. PMID- 9872477 TI - Apoptosis and antiphospholipid antibodies. AB - OBJECTIVE: To analyze the potential links between antiphospholipid antibodies (aPL) and apoptosis in the pathogenesis of the antiphospholipid antibody syndrome (APS). METHODS: A review was undertaken of the most relevant scientific literature on apoptosis and autoimmune phenomena. Experimental and human pathology were reviewed to substantiate the hypothesis that apoptosis is involved in the generation of aPL. RESULTS: Several considerations suggest that exposure of phospholipids (PL) during apoptosis may be a driving antigenic stimulus to the production of aPL. Furthermore, the molecular PL-protein complexes formed during apoptosis are targeted by "pathogenic" aPL. The binding and the clearance of apoptotic cells by these autoantibodies likely further enhances the aPL immune response. Experimental models and human pathology suggest that a restricted genetic background is key to the development of this immune response. CONCLUSIONS: Abnormalities of apoptosis observed in the course of autoimmune conditions likely provide an antigenic stimulus to the production of aPL. PMID- 9872478 TI - Internuclear ophthalmoplegia in systemic lupus erythematosus. AB - OBJECTIVES: To describe and review internuclear ophthalmoplegia (INO) in systemic lupus erythematosus (SLE). PATIENTS AND METHODS: A population of 268 SLE patients was retrospectively studied. INO was clinically defined as palsy of the ipsilateral rectus muscle and failure in contralateral eye adduction with dissociated nystagmus. A systematic review of the literature was made using MEDLINE (Silver-Platter) between 1966 and 1997, and for completeness, earlier references cited in identified articles. RESULTS: Four women with INO were identified. Their mean age at INO diagnosis was 38 years, and mean delay from diagnosis of SLE to INO was 6 years. INO was unilateral in all and coincided with disease activity in three. Cardiovascular risk factors were present in three. Magnetic brain resonance showed multiple and hyperintense (T2) lesions in white matter without correlation with clinical features. Other ancillary tests were not helpful for diagnosis. Corticosteroid therapy resulted in full resolution of INO in three cases. Review of 14 additional cases from the literature showed a similar experience. CONCLUSIONS: INO is uncommon in SLE, but it should be suspected in young patients with active disease and impairment of ocular movements. Diagnosis relies largely on clinical grounds. Neuroimaging is of little help. Steroid therapy seems effective in improving eye movements. PMID- 9872479 TI - Coexistent minocycline-induced systemic lupus erythematosus and autoimmune hepatitis. AB - OBJECTIVES: This study was performed to raise awareness among rheumatologists about two autoimmune disorders associated with long-term minocycline therapy that can coexist in the same patient. We provide an update on the occurrence of these disorders, their main characteristics, and the current knowledge of potential pathogenic mechanisms. METHODS: We searched the medical literature in English indexed in MEDLINE from 1966 through April 1998 for the term minocycline combined with each of the following: autoimmune hepatitis (AIH), chronic hepatitis, lupus, systemic lupus erythematosus (SLE), anti-myeloperoxidase (anti-MPO), arthritis, vasculitis, and toxicity. We also reviewed relevant references cited in the articles our search uncovered. RESULTS: We identified over 60 minocycline-induced cases of SLE and 24 cases of minocycline-induced AIH. Both autoimmune disorders coexisted in the same patient in 12 cases reported in the literature and in one case seen at our clinic. These 13 patients were characterized by symmetrical polyarthralgias/polyarthritis, elevated liver enzymes, and positive antinuclear antibodies (ANA); they also were generally anti-histone-negative, and only two patients had perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCA). After withdrawal of minocycline, their symptoms resolved, and abnormal laboratory results normalized or markedly improved. CONCLUSIONS: Although data on the actual prevalence of autoimmune disorders induced by minocycline are not available, numerous case reports or small series deal with such disorders. Probable pathogenic mechanisms for each of these conditions are discussed. PMID- 9872480 TI - Disseminated histoplasmosis in systemic lupus erythematosus: case report and review of the literature. AB - OBJECTIVES: To report a patient who developed both central nervous system systemic lupus erythematosus (SLE) and disseminated histoplasmosis and to review the literature regarding histoplasma infection in patients with SLE. METHODS: MEDLINE review of the medical literature published in English. RESULTS: Disseminated histoplasmosis occurs rarely in patients with SLE. The main risk factor is treatment with corticosteroids at doses of 20 mg/d or greater. Fever, dyspnea, pleurisy, and weight loss are typical presenting symptoms. The most commonly involved tissues are lung, liver, and bone marrow. In our patient, both SLE flare and disseminated histoplasmosis were present simultaneously. CONCLUSIONS: Opportunistic infection is an important complication of SLE and may be difficult to diagnose. Symptoms of infection may mimic those of a lupus flare, or conversely, symptoms may be masked by the use of corticosteroids. Fever, unexplained tissue involvement, atypical clinical patterns, and poor response to immunosuppressive therapy should alert the clinician to aggressively pursue evaluation of possible infection in patients with SLE. PMID- 9872481 TI - Cryoglobulinemia in primary Sjogren's syndrome: prevalence and clinical characteristics in a series of 115 patients. AB - OBJECTIVES: To determine the prevalence and nature of cryoglobulins in a large series of patients with primary Sjogren's syndrome (SS) and identify the clinical and immunologic features related to their presence. METHODS: In a cross-sectional study, we investigated 115 consecutive patients (107 women and eight men) with primary SS. All patients fulfilled four or more of the preliminary diagnostic criteria for SS proposed by the European Community Study Group in 1993. Serum cryoglobulinemia was measured in all patients. Serum samples were obtained at 37 degrees C, and cryoglobulinemia was estimated by centrifugation after incubation at 4 degrees C for 7 days. The type of cryoglobulinemia was identified by agarose gel electrophoresis and immunofixation. RESULTS: Cryoglobulins were detected in the sera of 18 (16%) of our patients with primary SS; most were IgMkappa monoclonal/IgG polyclonal. When compared with patients without cryoglobulins, those with cryoglobulins presented a higher prevalence of leukocytoclastic cutaneous vasculitis (56% v8%, P < .001), hypocomplementemia (75% v 2%; P < 0.001) and antibodies to hepatitis C virus (HCV) (47% v8%, P < .001). Liver involvement (clinical signs, biochemical features, or ultrasound/histological data of liver disease) was present in all patients (100%) with cryoglobulins and HCV infection but in only 11% of patients with cryoglobulins without HCV infection (P < .001). CONCLUSIONS: Leukocytoclastic cutaneous vasculitis, hypocomplementemia, and HCV infection are associated with the presence of cryoglobulins in the sera of patients with primary SS. Testing for HCV infection is recommended for patients with SS and cryoglobulinemia because of its high prevalence and its strong association with liver disease. PMID- 9872482 TI - Temporal arteritis and sensorineural hearing loss. AB - OBJECTIVES: The association of temporal arteritis and sensorineural hearing loss is a rare occurrence. We present four cases with both these disorders. METHODS: A 10-year retrospective review of all cases seen at the Geisinger Medical Center showed 271 cases of temporal arteritis and four cases with concomitant sensorineural hearing loss. The four cases were identified, obtained, and extensively reviewed. Patients ranged in age from 59 to 76 years and presented with sensorineural hearing loss and signs, symptoms, and biopsy evidence of temporal arteritis. The response to therapy and clinical course of each case are discussed. RESULTS: In three patients, the hearing loss preceded the diagnosis of giant cell arteritis by 2 days to 2 months. In one case, the hearing loss was noted concurrently with systemic features of giant cell arteritis. All cases were treated with corticosteroids and had full or partial return of hearing. CONCLUSIONS: Sensorineural hearing loss can be a preceding or concurrent symptom of temporal arteritis. Recognition and treatment may lead to partial or full recovery. It is not certain whether the time from onset of hearing loss to diagnosis to institution of therapy influences recovery. PMID- 9872483 TI - Women and menopause: beliefs, attitudes, and behaviors. The North American Menopause Society 1997 Menopause Survey. AB - OBJECTIVE: The main purpose in organizing this survey was to collect information relevant to The North American Menopause Society's (NAMS) educational mission and to document women's knowledge of, and attitudes toward, menopause. DESIGN: During June-July 1997, The Gallup Organization conducted 750 telephone interviews with a randomly selected sample of women 45-60 years of age from across the United States. Women were asked about their sources of information on menopause, what changes in health they anticipated as a result of menopause, why they used hormone therapy, and their attitudes toward menopause as a natural or a medical event. RESULTS: Women are more likely to believe that depression and irritability are associated with menopause than heart disease, but only a few associate menopause with an increasing vulnerability to either memory loss or Alzheimer's disease. Relief of physical symptoms of menopause was mentioned as the reason for starting hormone therapy more often than to protect against osteoporosis (25% relative to 15%), or to prevent stroke or a heart attack (10%), or to reduce the risk of developing Alzheimer's disease (2%). The single main source of women's information on menopause was a health professional (49%). The majority of women who were already menopausal or experiencing menstrual changes expressed an attitude toward menopause that was either neutral (42%) or positive (36%). CONCLUSIONS: Women are divided in their views of menopause, some seeing it as a medical condition requiring medical treatment, whereas others see it as a natural transition to be managed by "natural" means. Providing women with accurate, up-to date information and enhancing communication between healthcare providers and menopausal women remain the challenges for NAMS. PMID- 9872484 TI - The relationship of ovarian production of hormones and degree of ovarian stromal hyperplasia in postmenopausal women. PMID- 9872485 TI - Relationship between ovarian production of estrone, estradiol, testosterone, and androstenedione and the ovarian degree of stromal hyperplasia in postmenopausal women. AB - OBJECTIVE: To study the relationship between ovarian production of estrone (E ), estradiol (E2), testosterone (T), and androstenedione (A) and the ovarian degree of stromal hyperplasia in postmenopausal women. DESIGN: In 18 postmenopausal women, the ovarian vein hormone levels of E1, E2, T, and A were compared with the degree of ovarian stromal hyperplasia. The degree of stromal hyperplasia was assessed by histological analysis (group 1: atrophic ovaries, n = 8; group 2: slight stromal hyperplasia, n = 8; group 3: moderate or severe stromal hyperplasia, n = 2). RESULTS: The ovarian levels of E1 and E2 did not correlate with the ovarian degree of stromal hyperplasia. The ovarian levels of A in group 3 were significantly higher than those in groups 1 and 2 (p < 0.02 and p < 0.01, respectively). The ovarian levels of T in group 3 were significantly higher than those in group 1 (p < 0.01) but did not differ significantly from those in group 2. CONCLUSIONS: The amount of stromal hyperplasia in postmenopausal ovaries is correlated with the ovarian vein levels of A and T. Morphological characteristics of the postmenopausal ovary determine the local (pelvic) endocrine status and may play a role in the etiology of hormone-dependent diseases of the internal genitals. PMID- 9872486 TI - Raloxifene induces neurite outgrowth in estrogen receptor positive PC12 cells. AB - OBJECTIVE: It is well established that gonadal steroids have direct in vivo and in vitro effects on neurons. To further study these effects, we used rat PC 12 cells to examine the effects of estrogen receptor (ER) ligands on neuronal morphology. DESIGN: PC 12 cells constitutively express ER beta, but only strongly express ER alpha after long-term priming with nerve growth factor (NGF). We therefore primed PC12 cells with NGF for 14 days before testing them for estradiol (10(-9)M)- and/or raloxifene (10(-7) M)-induced neurite growth. Neurite growth was assessed by quantitative light microscopy. As control, ER status of the PC12 cells was assessed by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In this study, both estradiol and raloxifene induced the outgrowth of neurites in NGF-treated PC 12 cells (p < 0.05). The combination of estradiol- and raloxifene-induced neurite growth was statistically greater than the effects of either agent alone. RT-PCR confirms that NGF-treated PC 12 cells express both ERalpha and ERbeta. CONCLUSIONS: This report is the first on the neurotrophic effect of raloxifene. At 10(-7) M, raloxifene's effect equaled that of estradiol; moreover, raloxifene did not block the neurite growth of simultaneously estradiol-treated PC 12 cells, despite its functional antiestrogenic effects in vivo. We conclude that raloxifene is estrogen agonistic in this animal model and therefore studies are warranted to delineate the relationship between steroidal estrogen and raloxifene. PMID- 9872487 TI - Plasma insulin-like growth factor-I and its binding proteins 1 and 3 during continuous nonoral and oral combined hormone replacement therapy. AB - OBJECTIVE: We determined the effects of continuous combined nonoral and oral estrogen-progestin therapy on plasma levels of insulin-like growth factor-I (IGF I) and its binding proteins 1 (IGFBP-1) and 3 (IGFBP-3). DESIGN: Forty women complaining of climacteric symptoms were randomized to receive either transdermal estradiol patches (50 microg daily) in combination with an intrauterine system releasing 20 microg of levonorgestrel daily or an established regimen of an oral dose of 2 mg of estradiol and 1 mg of norethisterone acetate daily for 1 year. Fifteen women in the nonoral therapy group and 17 in the oral therapy group completed the 1-year prospective study. RESULTS: Plasma levels of IGF-I decreased significantly in the oral therapy group, but no changes were observed in the plasma levels of its binding proteins. Transdermal estrogen in combination with intrauterine levonorgestrel did not induce any change in plasma IGF-I, whereas the plasma levels of IGFBP-3 decreased. IGFBP-1 concentrations increased, which may be related to the induction of this protein into the endometrium by the intrauterine levonorgestrel delivery. CONCLUSIONS: Both the nonoral and oral continuous combined estrogen-progestin therapies used in this study produced only minor changes in the circulating concentrations of IGF-I and its binding proteins. PMID- 9872488 TI - Abnormal sonographic endometrial findings in asymptomatic postmenopausal women: possible role of antihypertensive drugs. AB - OBJECTIVE: The purpose of this study was to evaluate the possible effect of the antihypertensive agent nifedipine (a calcium channel antagonist) on the endometrium of asymptomatic menopausal women by using transvaginal sonography, hysteroscopy, and endometrial sampling. DESIGN: Eighty-five asymptomatic postmenopausal women with an ultrasonographically detected endometrial thickness of more than 7 mm were examined. The study group comprised 43 women treated with nifedipine (30-60 mg/day) for mild to moderate hypertension for at least 1 year, and the control group comprised 42 normotensive women. All participants underwent transvaginal sonography, hysteroscopy, and, if necessary, endometrial biopsy. RESULTS: Of the 85 patients, 53 were found to have atrophic endometrium, 36 (83.7%) in the study group and 17 (40.5%) in the control group. The difference between the groups was statistically significant (p < 0.05). There were three cases of hyperplasia and three cases of carcinoma of the endometrium, with similar rates in the two groups. CONCLUSIONS: Among asymptomatic postmenopausal patients with sonographically thickened endometrium, atrophic endometrium was found to be significantly more prevalent in those using calcium channel antagonists to control hypertension than in those who were normotensive controls. The presence of endometrial hyperplasia and malignancy in the study group underscores the importance of meticulous endometrial evaluation whenever sonographically thickened endometrium is encountered. PMID- 9872489 TI - Patient noncompliance with hormone replacement therapy: a nationwide estimate using a large prescription claims database. AB - OBJECTIVE: The purpose of this investigation was to use a large prescription claims database to characterize noncompliance with hormone replacement therapy (HRT). DESIGN: A retrospective longitudinal analysis was conducted among 28,718 women aged 40-59 years who were new HRT users. In the present study, HRT was defined as an oral estrogen (with or without progestin in separate prescribing) or as an oral estrogen combination with 21 or more recommended therapy days for every 28 calendar days. Compliance was determined by constructing a ratio of the total days of HRT received during the follow-up period divided by the total number of days in the follow-up period (365 days). An individual with a medication availability ratio less than 0.75 was considered to be noncompliant. RESULTS: After 1 year, 54.4% of the study subjects were noncompliant with HRT. Noncompliance was associated with younger age, living in the southern region of the United States, and not having an obstetrician/gynecologist as the initial prescriber. CONCLUSIONS: Noncompliance with HRT in the general population is higher than that reported in randomized clinical trials, suggesting that women are not benefiting optimally from therapy and that they are at risk for unnecessary morbidity and mortality from osteoporosis and cardiovascular disease. Increased efforts to improve long-term HRT compliance are urgently needed. PMID- 9872490 TI - Dietary fat and calcium intakes of menopausal women. AB - OBJECTIVE: The purpose of this retrospective, descriptive study was to assess the baseline dietary intake of fat and calcium in a group of women attending a multidisciplinary menopause clinic. DESIGN: Dietary fat and calcium intakes of this group were determined with food-frequency questionnaires, food records, and dietary interviews. The dietary fat and calcium intakes of those patients who expressed concern of risk for cardiovascular disease or osteoporosis and those patients using herbal preparations were compared with those who did not. RESULTS: Of 75 women, the majority of patients exceeded the dietary fat recommendations and had insufficient intakes of calcium. Significantly fewer patients with an expressed concern of risk for cardiovascular disease exceeded the recommendation for total fat and saturated fat (p < 0.05). There was no significant difference in calcium intake between those with and those without an expressed concern of risk for osteoporosis or in the diets of patients who did or those who did not take herbal products. CONCLUSIONS: The large proportion of patients not meeting recommendations for fat or calcium intakes confirms that the assessment of baseline dietary habits and nutrition education and intervention should be an essential component of the management of menopausal women. PMID- 9872493 TI - The role of epidermal growth factor (EGF) and its receptor in mucosal protection, adaptation to injury, and ulcer healing: involvement of EGF-R signal transduction pathways. AB - Growth factors and their receptors are known to play important roles in normal cell proliferation, morphogenesis, tissue repair, and ulcer healing. Epidermal growth factor (EGF) inhibits acid secretion, exerts a trophic effect on gastroduodenal mucosa, protects gastric mucosa against injury, mediates mucosal adaptation, and accelerates gastroduodenal ulcer healing by stimulating cell migration and proliferation. EGF exerts its actions by binding to its receptor, EGF-R, a transmembrane protein tyrosine kinase, which triggers receptor dimerization, autophosphorylation, and recruitment of kinase substrates. These events result in Ras (GTP-binding protein) activation of the Ras/Raf/MAP kinase pathway, leading to phosphorylation of regulatory proteins and transcription factors and culminating in cell proliferation. Other pathways potentially activated by EGF include the phosphatidylinositol pathway and the JAK/STAT signaling pathway. Recent studies demonstrated that EGF-R-associated tyrosine kinase plays an essential role in regulating gastric mucosal cell proliferation after acute injury and further demonstrated activation of the EGF-R gene, EGF-R phosphorylation, and increased MAP kinase activity during early stages of experimental gastric ulcer healing. Finally, experimental data indicate that Helicobacter pylori vacuolating cytotoxin inhibits healing of experimental gastric ulcers, cell proliferation, binding of EGF to its receptor, EGF-induced EGF-R phosphorylation, and MAP kinase (ERK-2) activation. These H. pylori actions can explain its interference with the ulcer healing process. PMID- 9872492 TI - Prostaglandins in the stomach: an update. AB - Prostaglandins (PGs) are responsible for regulation of various physiologic activities in many tissues and organs, including the stomach. Recent studies have shown new crucial roles of PGs in the stomach. Activation of inflammatory cytokines and neutrophils may cause acute gastric mucosal lesions and recurrence of ulcers, which are induced by noxious stimuli such as nonsteroidal antiinflammatory drugs (NSAIDs), stress and Helicobacter pylori (H. pylori). These phenomena are PG-dependent because exogenous PGs reverse them. PG deficiency and H. pylori may worsen the quality of ulcer healing in terms of inflammatory responses, which are related to future ulcer recurrence. Neutrophils, monocytes/macrophages, and adhesion molecules are involved in the mechanism of recurrence caused by inflammatory cytokines. PGs accelerate ulcer healing, possibly via angiogenesis, epithelial cell proliferation, production of growth factors such as hepatocyte growth factor and transforming growth factor beta, reconstruction of extracellular matrices, and suppression of inflammatory cell infiltration, in addition to gastroprotective mechanisms. The PG synthase cyclooxygenase (COX) has two forms, COX-1 and COX-2. COX-2, but not COX-1, contributes to ulcer healing. Moreover, recent studies suggest the involvement of COX-2 in development of gastric and colon carcinoma. This may be linked to the chemopreventive effect of NSAIDs. PMID- 9872494 TI - Induction of cyclooxygenase-2 in a rat gastric epithelial cell line by epiregulin and basic fibroblast growth factor. AB - Prostaglandins play an important role in maintaining gastric mucosal integrity. Cyclooxygenases (COX-1 and -2) are the key enzymes involved in prostaglandin synthesis. COX-2 expression in gastric epithelial cells remains a subject of controversy, and a possible regulation of gastric COX-2 by growth factors has not been explored. Therefore, we studied the effect of growth factors including epiregulin, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) on expression of COX-2 in a gastric epithelial cell line (RGM1) derived from normal rat gastric mucosa. Cells were incubated with 10 or 100 ng/ml of EGF. epiregulin, bFGF, or VEGF for 1, 2, 3, 6, and 24 h. COX-2 mRNA expression was determined by RT-PCR using specific COX-2 primers and COX-2 protein expression was determined by Western blotting. This study showed that COX 2 mRNA and protein are expressed in the gastric epithelial RGM1 cell line and that epiregulin and bFGF (but not VEGF) significantly increase expression of COX 2 mRNA and protein. Because PGs play an important role in mucosal defense, this study suggests that some growth factors contribute to maintaining mucosal integrity via activation of the COX-2 gene. PMID- 9872495 TI - Selective inhibitors of cyclooxygenase-2: are they really effective, selective, and GI-safe? AB - Selective inhibitors of the "inducible" isoform of cyclooxygenase (COX-2) have been suggested to be effective analgesic and anti-inflammatory drugs while sparing the gastrointestinal (GI) tract of injury. There is some experimental and early clinical evidence to support this hypothesis. However, some important questions remain regarding the utility of selective COX-2 inhibitors. For example, estimates of the selectivity of COX-2 inhibitors based on in vitro studies are likely to be poor predictors of selectivity in vivo. Efficacy with selective blockade of COX-2 may be inferior to that achieved with combined inhibition of COX-1 and COX-2. Furthermore, in situations in which there is inflammation or ulceration in the GI tract, COX-2 produces prostaglandins that are essential for repair. In these circumstances, inhibition of COX-2 leads to delay of ulcer healing and exacerbation of inflammation. Some caution should therefore be exercised before the theory is fully accepted that selective COX-2 inhibitors are effective anti-inflammatory drugs that spare the GI tract of injury. PMID- 9872496 TI - A virulence factor of Helicobacter pylori: role of heat shock protein in mucosal inflammation after H. pylori infection. AB - Among the various virulence factors of Helicobacter pylori the role of its heat shock protein 60 (HSP60, HspB) in mucosal inflammation after H. pylori infection was examined. In flow cytometric analysis, the expression of HSP60 on the cell surface was different, depending on the H. pylori strain used. The HSP60 epitope was also detected on the surface of both human gastric cancer cells (MKN45, KATOIII, and MKN28) and human gastric biopsy specimens. The intensity of the expression of HSP60 on the cell surface correlated significantly with the adhesion of H. pylori to MKN45 cells, but not with urease activity and production of vacuolating cytotoxin. A monoclonal antibody to H. pylori HSP60 inhibited the adhesion of H. pylori to MKN45 cells. These results suggest that HSP60 of H. pylori might act as an important virulence factor after H. pylori infection. PMID- 9872497 TI - Role of cyclooxygenase 2 in hepatocyte growth factor-mediated gastric epithelial restitution. AB - Migration of epithelial cells (restitution) is an essential step in the repair of gastric mucosal lesions. Although a variety of growth factors are reported to facilitate gastric epithelial restitution, the intracellular mechanisms of this process are not fully understood. In this study we investigated the effects of hepatocyte growth factor (HGF) on restitution of normal rat gastric epithelial RGM-1 cell monolayers after injury and examined whether cyclooxygenase-2 (COX-2) is involved in HGF-mediated epithelial restitution. Restitution of RGM-1 monolayers was assessed using a round wound restitution model. Application of HGF (5 ng/ml) significantly facilitated the restitution of RGM-1 monolayers after artificial wounding. HGF also induced expression of COX-2 protein in RGM-1 cells, and wounding itself induced COX-2 expression in the cells located at the edge of the wound. Inhibition of COX-2 activity by NS-398, a specific COX-2 inhibitor, significantly delayed the HGF-mediated restitution. These results suggest the involvement of COX-2 in the action of HGF on gastric epithelial restitution. PMID- 9872498 TI - Effects of NSAIDs on proliferation of gastric cancer cells in vitro: possible implication of cyclooxygenase-2 in cancer development. AB - The roles of cyclooxygenase-2 (COX-2) in the development of gastric cancer are unknown. We investigated the effects of nonsteroidal antiinflammatory drugs (NSAIDs), which are specific and nonspecific inhibitors of COX-2, on proliferation of the gastric cancer cell lines KATOIII, MKN28, and MKN45. The protein level of COX-2 was examined in these cell lines by Western analysis, and mRNA levels of COX-1/2 by Northern analysis. These cell lines expressed comparable levels of COX-1 mRNA. However, mRNA and protein expression of COX-2 in these cell lines was different. MKN45 expressed higher levels of COX-2 mRNA and protein than KATOIII and MKN28. We also examined the effects of NS-398 and indomethacin, specific and nonspecific inhibitors of COX-2, on the increase in cell number and [3H]thymidine uptake of these cell lines. NS-398 and indomethacin suppressed proliferation of MKN45 cells that overexpressed COX-2, although they exerted minimal effects on proliferation of KATOIII and MKN28, which expressed lower levels of COX-2. These results are consistent with the hypothesis that COX 2 is expressed in certain groups of gastric cancers and is related to their cell proliferation. It was proposed that COX-2 plays an important role in development of gastric cancer cells. Furthermore, NSAIDs may exert antiproliferative activity against gastric adenocarcinomas that overexpress COX-2. PMID- 9872499 TI - Gastric cytoprotection is secondary to increased mucosal fluid secretion: a study of six cytoprotective agents in the rat. AB - We tested the hypothesis that rapidly developing gastric cytoprotection produced by topical application of exogenous compounds is a result of increased gastric mucosal fluid secretion. Ex vivo gastric chambers were prepared in rats which were subsequently exposed topically to one of the prostaglandin (PG) E1 analogues misoprostol or rioprostil, PGE2, nicotine, N-ethylmaleimide (NEM), 0.25 M HCl, or to their respective vehicles. All agents were added to empty chambers to avoid complications resulting from dilution by gastric contents. Effects of these agents on intraluminal volume changes, blood flow, juxtamucosal pH, histology, and on the mucosal damage resulting from necrotizing agents were studied. All six agents were cytoprotective and each increased net secretion of fluid by the chambered mucosae. Gastric blood flow was not significantly increased by NEM, by 0.25 M HCl, or by nicotine compared to controls, and the juxtamucosal pH was not significantly increased by any of the three agents for which this was studied. Vacuole formation in surface epithelial cells and subepithelial edema were seen after exposure to some agents, but none of the agents led to formation of a thick barrier of exfoliated cells and mucus. Ablation of primary afferent nerves with capsaicin abolished both protection by 0.25 M HCl and the net increase in fluid secretion by the mucosae. Capsaicin ablation did not alter either the protection afforded by NEM or the increase in volume of secretion. We conclude that increased mucosal fluid secretion is the common factor present with all six cytoprotective agents and hence may be the predominant mechanism of cytoprotection against topically applied necrotizing agents. PMID- 9872500 TI - Inhibition of inducible nitric oxide synthase delays gastric ulcer healing in the rat. AB - We sought to clarify the role of nitric oxide (NO) generated from inducible NO synthase (iNOS) during healing of rat gastric ulcers. After gastric ulcers were induced by acetic acid, rats were treated with vehicle, N(G)-nitro-L-arginine methyl ester (L-NAME), aminoguanidine (AG), and dexamethasone (Dex) by gastric intubation twice a day for 3 days to 1 week. L-NAME significantly delayed healing compared with vehicle. AG and Dex significantly reduced ulcer size 3 days after ulcer induction but did not further reduce ulcer size 1 week after induction. iNOS expression was present in inflammatory cells, some epithelial cells, and in vascular smooth muscle in the regenerating mucosa of the vehicle-treated group. However, the number of iNOS-positive inflammatory cells increased in the AG- and L-NAME-treated groups. AG and L-NAME significantly increased the number of inflammatory cells with endogenous peroxidase and significantly reduced the number of apoptotic inflammatory cells compared with vehicle. In conclusion, inhibition of iNOS increases the number of inflammatory cells in the ulcer margin and delays ulcer healing. These observations suggest that NO generated from iNOS not only participates in ulcer formation but also plays a beneficial role in ulcer healing, in part by the exclusion of iNOS-positive inflammatory cells from the regenerating mucosa. PMID- 9872501 TI - Activation of microvascular endothelial cells in active ulcerative colitis and detection of inducible nitric oxide synthase. AB - Endothelial nitric oxide (NO) synthase (NOS) that is expressed constitutively on microvascular endothelium is believed to be essential to systemic and/or local vascular integrity. Endothelial cells (ECs) were reported to express inducible NOS (iNOS) under some conditions iNOS expression indicates vascular activation. In this study we examined microvascular activation using ECs obtained from patients with ulcerative colitis (UC). We cultured ECs from the mesenteries of surgical UC patients and assayed NOS activity by NADPH-diaphorase cytochemistry and immunocytochemistry with an anti-iNOS antibody. Strong NOS activity was demonstrated on the cells from UC patients (5/5), whereas no activity was detected on the cells from cancer patients and human umbilical vein endothelial cells (HUVEC 0/5, 0/5). Strong iNOS activity was detected by immunoreaction, and large amounts of NO generated were detected by the conversion from [14C]arginine to [14C]citrulline (HUVEC 624+/-376 vs. EC-UC 1,492+/-233 dpm). These results suggest the possibility that ECs express spontaneous and continuous iNOS in active UC. They indicate a close relationship of vascular activation with the pathogenesis of UC. PMID- 9872502 TI - Effects of cigarette smoking on gastric ulcer formation and healing: possible mechanisms of action. AB - Epidemiologic studies have shown that cigarette smoking is closely related to peptic ulcer disease. The mechanisms by which cigarette smoking adversely affects gastric mucosa have been suggested and elucidated. This article reviews some of the mechanisms involved in cigarette smoking-related gastric ulceration and healing. Experimental findings suggest that cigarette smoking increases xanthine oxidase activity, leukotrienes, and nitric oxide production and also neutrophil infiltration in the gastric mucosa. On the other hand, it reduces blood flow, prostaglandin production, epithelial cell proliferation, and formation of blood vessels in the tissue. These actions are important for ulcer formation and healing. The evidence thus far available strengthens the hypothesis that cigarette smoke is indeed harmful to gastric mucosa through defined mechanisms. PMID- 9872503 TI - Molecular mechanisms involved in NSAID-induced gastropathy. AB - A growing body of experimental evidence indicates that leukocyte-endothelial cell interactions may play an important role in the pathogenesis of NSAID-induced gastropathy. Using a newly described, dual radiolabeled monoclonal antibody technique to quantify adhesion molecule surface expression in vivo, we have demonstrated increases in surface expression of ICAM-1 and P-selectin in the gastric mucosa after oral administration of indomethacin. We have also found that CD18-, ICAM-1-, or P-selectin-deficient mice are less sensitive to the ulcerogenic effects of orally administered indomethacin. Although there is virtually no information regarding the regulation of expression of endothelial cell adhesion molecules (ECAMs) in experimental NSAID-induced gastropathy, the nuclear transcription factor KB (NFKB) may represent a potential modulator of transcriptional activation of ECAM expression. We have demonstrated that two structurally distinct yet highly selective proteasome inhibitors (MG341, lactacystin) inhibit tumor necrosis factor (TNF)-induced NFKB activation as well as ECAM expression in human endothelial cells in vitro. In addition, we found that these proteasome inhibitors significantly reduced indomethacin-induced gastric mucosal injury as well as gastric mucosal ICAM-1 expression in the rat in vivo. We conclude from these studies that indomethacin activates NFKB (possibly via TNF synthesis) in gastric microvascular endothelial cells, thereby enhancing surface expression of ICAM-1 which binds the CD18 on polymorphonuclear leukocytes (PMNs). These adherent PMNs are then believed to mediate endothelial and/or epithelial cell injury either directly or indirectly. PMID- 9872504 TI - Downregulation of gastric mucin gene expression and its biosynthesis by dexamethasone in the human. AB - The effect of corticosteroids on the release and biosynthesis of gastric mucin remains unclear. We studied the effects of dexamethasone on biosynthesis of mucin and MUC1 gene expression in the human stomach in vitro. Gastric mucosal specimens, obtained from six subjects at gastrectomy, were cultured with various concentrations of dexamethasone. Biosynthesis of mucin was studied by labeling gastric mucosa for 2 h with [3H]-glucosamine. After purification of mucin by CsCl gradient centrifugation, radioactivity of intra- and extracellular samples was counted. MUC1 gene expression was studied by Northern and dot-blot analysis using a cDNA encoding human gastric mucin gene MUC1. The dexamethasone treatment decreased mucin secretion from the isolated mucosa in a time- and concentration dependent manner, with maximal inhibition of secretion (32+/-6%) observed after 8 h and 10(-5) M. Dexamethasone treatment also decreased MUC1 levels (28+/-9%). The inhibitory effect was also observed in carbachol-evoked secretion. These findings suggest that a decrease in mucin biosynthesis by corticosteroids may be involved in steroid-induced gastric mucosal damage. PMID- 9872505 TI - Polyamines and epidermal growth factor in the recovery of gastric mucosa from stress-induced gastric lesions. AB - Polyamines such as spermine or putrescine, resulting from increased activity of ornithine decarboxylase (ODC), are known for gastroprotective and mucosal growth promoting effects. EGF exhibits similar effects, but little is known about the involvement of polyamines in acceleration of the healing of stress-induced gastric lesions by epidermal growth factor (EGF). In this study, rats with intact or suppressed ODC activity by alpha-difluoromethy-ornithine (DFMO, 400 mg/kg i.p.) were subjected to 3.5 h of water immersion and restraint stress (WRS) without or with addition of spermine or EGF. At 0, 2, 6, 12, or 24 h after stress, rats were sacrificed. The number of gastric lesions was determined and gastric blood flow (GBF) was recorded by the H2 gas clearance technique. Stress produced gastric lesions (mean number 18+/-2 per stomach) and decreased GBF (by approximately 43%), but at 2, 6, 12, and 24 h after stress, these lesions and the decrease in GBF were gradually attenuated. Pretreatment with DFMO or removal of an endogenous source of EGF by salivectomy resulted in a marked decrease in mucosal DNA synthesis and significantly delayed the healing of stress lesions. EGF or spermine significantly accelerated ulcer healing and increased the GBF in rats with intact or removed salivary glands. DFMO significantly reduced the enhancement of healing and the increase in GBF induced by EGF, but failed to influence those induced by exogenous spermine. We conclude that polyamines play an important role in mucosal recovery from stress lesions due to acceleration of mucosal repair and increase in gastric microcirculation and that increased ODC activity and resulting excessive polyamine release appear to act as primary mediators of EGF-induced acceleration of the healing of stress lesions. PMID- 9872506 TI - A neurotrophic pyrimidine compound, MS-818, enhances EGF-induced restoration of gastric epithelial wounds in vitro. AB - MS-818 is a novel synthetic pyrimidine compound that stimulates nerve regeneration and promotes synthesis of various growth factors and differentiation of astrocytes. Effects of MS-818 on gastric epithelial cells were assessed using a wound repair model with primary cultured gastric epithelial cells from rabbits. A round wound with a constant cell-free area was created and the process of restoration was monitored by measuring wound size every 12 h. Cell proliferation was monitored by sequential staining with BrdU. As previously reported, EGF (10 ng/ml) accelerated wound repair by promoting cell migration and proliferation. Although MS-818 alone had no effects, MS-818 (10-100 microM) enhanced EGF-induced acceleration of gastric epithelial restoration, including cell migration and proliferation. Although the detailed mechanism of action of this agent is still unclear, MS-818 might have favorable effects on in vivo gastric mucosal repair. PMID- 9872507 TI - Protective role of melatonin and the pineal gland in modulating water immersion restraint stress ulcer in rats. AB - We investigated the protective effect of melatonin on stress-induced gastric lesions in rats. Fasted rats were subjected to water immersion restraint stress for 4 h and the percentage of corpus mucosa containing hemorrhagic lesions was determined. Thirty minutes before restraint stress, melatonin or vehicle was administered i.p. In another experiment, pinealectomy was performed 1 week before water immersion restraint stress. Administration of melatonin at 1 and 5 mg/kg significantly decreased gastric lesions by 46 and 74%, respectively. In contrast, pinealectomy significantly enlarged the lesion area, although this effect was counteracted by melatonin at a dose of 1 mg/kg i.p. However, this protective effect of melatonin was abolished by i.p. pretreatment with indomethacin at 5 mg/kg. These results suggest that melatonin has gastroprotective properties against stress-induced gastric injury in rats and that the pineal gland contributes to gastric protection via prostaglandin-dependent mechanisms. PMID- 9872508 TI - Gastrin-regulated expression of p53 in transformed enterochromaffin-like cells in the African rodent mastomys. AB - The tumor suppressor p53 functions at the G1/S-phase checkpoint of the cell cycle to direct cells that have accumulated somatic mutations toward apoptosis and away from mitosis. The p53 gene is commonly mutated in human cancers, but the molecular mechanisms regulating this event are not clear. The African rodent mastomys exhibits a genetic predisposition to develop gastric carcinoids derived from enterochromaffin-like (ECL) cells. The ECL cell transformation can be accelerated by acid inhibition-induced hypergastrinemia. This study evaluates the alteration of p53 during the rapid ECL cell transformation. Hypergastrinemia was generated by the irreversible histamine-2 receptor antagonist loxtidine for 8 weeks (hyperplasia) and 16 weeks (neoplasia). p53 expression was evaluated in fundic mucosa from different stages of transformation by Western blot analysis and immunohistochemistry using monoclonal antibodies against wild-type p53. RT PCR and molecular sequence analysis of p53 were undertaken with mRNA isolated from purified ECL cells. Overproduction of the wild type of p53 was evident in ECL cells during hypergastrinemia, and the molecular characteristics of p53 were determined in naive and transformed ECL cells. p53 was mutated at the C-terminus in ECLoma induced by hypergastrinemia. Therefore, p53 is altered from overproduction to mutation during the development of hypergastrinemia-induced ECLoma and it may therefore play a role in the cell transformation. PMID- 9872509 TI - Involvement of endogenous cholecystokinin and somatostatin in gastroprotection induced by intraduodenal fat. AB - Duodenal fat such as oleate is known to influence gut functions by release of cholecystokinin (CCK), but the contribution of CCK endogenously released by duodenal fat or by diversion of pancreatic juice from the duodenum in the mechanism of mucosal integrity and gastroprotection has been little studied. This study was designed to compare the effect of CCK-8 and intraduodenal (i.d.) instillation of sodium oleate, or diversion of the pancreatic biliary secretions that are known to release CCK, on the gastric mucosal lesions induced by topical application of 100% ethanol or acidified aspirin (ASA) in rats with or without the pretreatment with a CCK-A receptor antagonist, loxiglumide, or with L-365,260 to block CCK-B receptors. In addition, the effect of suppression of prostaglandin (PG) biosynthesis by indomethacin (5 mg/kg i.p.), inhibition of nitric oxide (NO) synthase by L-NAME (5 mg/kg i.v.), or blockade of sensory nerves by capsaicin (125 mg/kg s.c.) on the protective activity of sodium oleate was determined. Sodium oleate (50-200 mM i.d.), or diversion of pancreatic juice from the duodenum for 3 h that produced significant rise in plasma CCK levels, significantly reduced gastric lesions induced by 100% ethanol to an extent similar to that induced by exogenous CCK-8 (5 nmol/kg s.c.). The protective effect of oleate or diversion of pancreatic juice was accompanied by an increase in gastric blood flow (GBF). Both protection and accompanying hyperemia were completely abolished by blockade of CCK-A receptors with loxiglumide, whereas L 365,260, an antagonist of CCK-B receptors, had no effect. Oleate given i.d. significantly attenuated acidified ASA-induced gastric lesions and gastric secretion while increasing the luminal concentration of somatostatin. These effects were significantly reduced by loxiglumide but not by L-365,260. In contrast, CCK-8, which stimulated gastric acid secretion, failed to affect the lesions induced by acidified ASA and the decrease in the GBF produced by this ulcerogen. Indomethacin, which suppressed PG generation by approximately 90%, failed to influence the protective activity of oleate or CCK-8 against ethanol induced lesions, whereas L-NAME, vagotomy, or sensory denervation significantly attenuated this protection and accompanying hyperemia. Addition to L-NAME of L arginine, but not D-arginine, restored the protective and hyperemic effects of CCK-8 and duodenal oleate against gastric lesions induced by ethanol or acidified ASA. We conclude that endogenous CCK released by oleate or diversion of pancreatic secretion exerts a potent gastroprotective action on the stomach involving predominantly CCK-A receptors and depending on vagal activity, and hyperemia mediated by NO and sensory nerves but unrelated to acid secretory effects and endogenous PG. PMID- 9872510 TI - Gastric epithelial cells stimulate Helicobacter pylori growth. AB - Helicobacter pylori infects only human gastric epithelium, causes gastritis, and is strongly associated with gastroduodenal ulceration and gastric cancer. Colonization of the stomach with H. pylori is accompanied in the acute stage by an increased number of neutrophils in the lamina propria, indicative of gastric inflammation. It is interesting that H. pylori colonizes specifically human gastric-type epithelial cells. We studied whether the presence of gastric epithelial cells influenced H. pylori growth. H. pylori (NCTC 11637) was cultured on Skirrow agar with 7% horse blood. Kato-III cells, a human gastric cancer cell line, were cultured with RPMI 1640 plus 10% fetal bovine serum (FBS). Kato-III cells (10(5)/ml) were cultured with/ without H. pylori (10(8) cfu/ml) with RPMI 1640 + 1% FBS for 3 days. The number of Kato-III cells was counted with a hemacytometer. H. pylori with/without Kato-III cells was cultured with RPMI 1640 + 10% FBS for 2 hours, and plated on Skirrow agar. After 3 days we counted the number of H. pylori colonies. To detect the H. pylori colonies, we used a colony hybridization method. DNA of colonies was transferred to positively charged nylon membrane and hybridized by PCR with Hpl (5'-CTG-GAG-AGA-CTA-AGC-CCT-CC-3') and Hp2 (5'-ATT-ACT-GAC-GCT-GAT-TGT-GC-3')-amplified primers. We previously reported that the number of Kato-III cells was significantly decreased by co-incubation with H. pylori. The number of H. pylori colonies was significantly increased by coincubation with Kato-III cells. We conclude that the presence of human gastric epithelial cells is important for the growth of H. pylori. PMID- 9872511 TI - Pathologic changes in the glandular stomach and duodenum in an H. pylori-infected Mongolian gerbil model. AB - We have established a Helicobacter pylori-infected Mongolian gerbil model following Hirayama's method to investigate gastric diseases associated with H. pylori infection. We orally administered the culture broth of H. pylori ATCC 43504 to 8-week-old male Mongolian gerbils. After this, the gerbils were fed in a vinyl isolator. Subsequently, over the course of 48 weeks some of them were sacrificed for histopathologic examination and H. pylori culture. H. pylori colonization in the glandular stomach was seen in all the infected gerbils but only a few H. pylori were detected histologically. Acute inflammation, immature epithelium, and erosion were observed 2 weeks after H. pylori infection. Chronic inflammation was noted from 4 weeks after H. pylori infection. In addition, we found intestinal metaplasia and gastric ulcers from 12 and 24 weeks, respectively. There was mild to moderate inflammation in the duodenum but no ulcerative lesions or gastric metaplasia were observed. Some histologic findings were similar to those in humans, but inflammation occurred mainly in the deep mucosa and submucosa. This is a good animal model for H. pylori-associated gastric diseases but not for duodenal ulcers or gastric metaplasia. It might be useful for investigating the pathogenesis of H. pylori infection in the stomach. PMID- 9872513 TI - [13C]-urea breath test for assessment of cure of Helicobacter pylori infection at 1 month after treatment. AB - Although the [13C]-urea breath test has been reported to provide high specificity and sensitivity, its reliability in determination of cure, including when and how the assessment of cure is made, is not fully established. Here we evaluate the reliability of judgment of cure after 1 month of therapy using the [13C]-urea breath test, including the appropriate time to assess its cure. Our subjects were 199 H. pylori-infected patients (144 men and 55 women; 50.5+/-0.76 years) who underwent cure therapy, took the breath test at 1 month, and underwent both the breath test and invasive tests using endoscopy at 6 months after completion of therapy. Accuracy of the breath test in judgment of cure at 1 month was investigated by comparing the results at 1 month with those at 6 months when cut off value of the breath test was set at 5 per thousand. Chronologic alterations of the delta13C value were also analyzed in cured patients who underwent the breath test at 1, 3, and 6 months. In 167 (94.9%) of 176 patients whose breath test values were less than 5 per thousand, cure was reconfirmed at 6 months. In 14 (60.9%) of 23 patients whose values were greater than 5 per thousand, cure was demonstrated at 6 months. The mean delta13C values of 33 cured patients who took the breath test at 1 month and repeated the breath test at 3 and 6 months for any reasons were 4.14+/-0.74 per thousand, 2.02+/-0.23 per thousand, and 1.72+/-0.19 per thousand, respectively, and the value at 3 months was significantly smaller than that at 1 month. The reliability of assessment of cure by only the breath test at 1 month was sufficiently high (94.9%) once the patients were judged to be cured. However, because the breath test at 1 month yielded a high false-positive rate and the breath test values at 3 and 6 months were similar, evaluation of cure of H. pylori infection by the [13C]-urea breath test should preferably be done at 3 months after treatment to minimize false-positive results, or the concept of a borderline group should be established for patients with positive but relatively high delta13C values (5-10 per thousand) who repeated the breath test 3 months after treatment. PMID- 9872512 TI - Mucosal macrophage inflammatory protein-1alpha levels are increased in Helicobacter pylori infection. AB - We examined the relationship between the levels of macrophage inflammatory protein 1alpha (MIP-1alpha) and interleukin-8 (IL-8) in organ cultures of antral mucosal tissues, background gastroduodenal diseases, and grades of histologic gastritis. Significantly higher levels of MIP-1alpha and IL-8 were detected in patients with H. pylori infection than in those without infection. In H. pylori positive patients, mucosal specimens from patients with peptic ulcer disease showed higher levels of MIP-1alpha and IL-8 than the specimens obtained from patients with erosive gastritis or those from endoscopically normal mucosa, and this was particularly pronounced in patients with duodenal ulcer. There were positive correlations between MIP-1alpha and IL-8 levels and histologic grades of activity, inflammation, and H. pylori density as defined by the Sydney system. However, the degree of association with the inflammatory cell count was different between these two chemokines. MIP-1alpha levels had a stronger association with mononuclear cells than with neutrophils, whereas IL-8 levels showed an association with neutrophils and mononuclear cells to an almost equal degree. These results suggest that MIP-1alpha and IL-8 may play important roles as inflammatory mediators in the pathogenesis of histologically proven H. pylori associated gastritis. PMID- 9872514 TI - Helicobacter pylori infection in the remnant stomach after gastrectomy: with special reference to the difference between Billroth I and II anastomoses. AB - Helicobacter pylori infection is associated with many gastric diseases, such as peptic ulcer and gastric cancer. We examined the remnant stomach for H. pylori infection after gastrectomy for gastric cancer or peptic ulcer between October 1992 and July 1997. H. pylori DNA in the gastric juice of 109 patients [mean age 62.4 years, male/female 78/31, gastrectomy for gastric cancer 83/peptic ulcer 26, Billroth I (BI) anastomosis 72/Billroth II (BII) 37, mean postoperative interval 6.0 years] was amplified by PCR and detected by Southern blot hybridization. The serum of 135 patients was assayed by ELISA for IgG antibody against H. pylori (mean age 61.8 years, male/female 99/36, gastrectomy for gastric cancer 111/peptic ulcer 24, BI anastomosis 93/BII 42, mean postoperative interval 5.4 years). H. pylori was positive in 68/109 (62.4%) by PCR and 113/ 135 (83.7%) by ELISA. H. pylori cytotoxin gene cagA, a H. pylori virulence factor gene, was found in 15/16 (93.8%) cases by PCR. A significant difference in H. pylori positivity by PCR was found according to the type of anastomosis (BI vs. BII) but not according to age group, sex, disease (cancer or ulcer), or postoperative interval by PCR and ELISA. BII anastomosis was followed by a significantly lower rate of H. pylori infection (17/37; 45.9%) than BI anastomosis (51/72; 70.8%; p=0.01) according to the results of PCR. Moreover, some patients with BII anastomosis (3/8; 37.5%) showed positive to negative seroconversion for H. pylori infection after the operation (mean 2.47 years) according to the results of ELISA, but this phenomenon was not observed in patients with BI (0/12) anastomosis. This may reflect the role of bile reflux, which is more common in BII than BI, because bile reflux interferes with colonization by H. pylori. PMID- 9872515 TI - Relationship between the eradication of Helicobacter pylori and the healing pattern of peptic ulcer. AB - We investigated the relationship between the eradication of Helicobacter pylori and the stage after the eradication of this bacterium. Eighty-six patients with H. pylori who had gastric ulcer (n=45) or duodenal ulcer (n=41) were enrolled in the study. As eradication therapy, patients received 1,500 mg amoxicillin, 400 mg clarithromycin, and 30 mg lansoprazole for 2 weeks, followed by 30 mg lansoprazole for 6 weeks in patients with gastric ulcer or for 4 weeks in those with duodenal ulcer. The ulcer stages were evaluated using the indigocarmine method at the third and sixth month after entry. The overall eradication rate of H. pylori was 85% in this study. In the gastric ulcer group, 62% of H. pylori eradicated patients and 37.5% of H. pylori-uneradicated patients showed the S2 stage (white scar) at the sixth month. In the duodenal ulcer group, 61% of H. pylori-eradicated patients showed the S2 stage and none of the H. pylori uneradicated patients showed the S2 stage (p < 0.05). We concluded that H. pylori eradication might promote not ulcer healing but maturity of the ulcer scar, especially in duodenal ulcer patients, and that this might also be related to the reduction of ulcer relapse. PMID- 9872516 TI - Helicobacter pylori infection, chronic gastritis, and proton pump inhibitors. AB - The proton pump inhibitors (PPIs) omeprazole, lansoprazole, and pantoprazole are widely used as antisecretory drugs and, in association with antibiotics, for the treatment of Helicobacter pylori infections. PPIs possess antibacterial activity against H. pylori in vitro, and may also exert an anti-inflammatory effect by interfering with the cellular immune response to infection. Their antimicrobial activity is selective for H. pylori. Lansoprazole is the most effective, although its bactericidal activity is similar to that of omeprazole. Pantoprazole is the least effective. The mechanisms that account for the antibacterial effects of PPIs may depend on a structural similarity of PPIs to antibiotics which are active against H. pylori, on the inhibition of bacterial urease exerted by PPIs, or on the possible interaction of PPIs with bacterial ATPases that regulate the transmembrane ion flux. Recent studies have shown that PPIs have anti inflammatory actions and can interfere with the host-bacteria interactions. Lansoprazole can bind to polymorphonuclear leukocytes that infiltrate the gastric mucosa colonized by H. pylori and can thus inhibit the oxidative burst of activated inflammatory cells. In an in vivo study, lansoprazole reduced the degree of activity of histologic gastritis independently of the presence of H. pylori. In another study, omeprazole was capable of inhibiting the cytotoxic activity of NK T cells. Investigation of PPI interactions with H. pylori activities and the cellular immune response to the infection may help us to understand the pathogenic mechanisms of H. pylori-associated diseases and enable clinicians to better treat them. PMID- 9872517 TI - Effect of combined administration of lansoprazole and sofalcone on microvascular and connective tissue regeneration after ethanol-induced gastric mucosal damage. AB - We undertook the present study to clarify the alteration of localization of basic fibroblast growth factor (bFGF), endothelial cells, and myofibroblasts in the healing of ethanol-induced gastric mucosal damage by the combined administration of lansoprazole and sofalcone. Wistar strain male rats were used. Ethanol 50% was given through orogastric intubation. Thirty minutes later, an aqueous solution of lansoprazole, sofalcone, a combination of lansoprazole and sofalcone, or physiologic saline was given orally. The stomach was removed and the localization of bFGF, myofibroblast, and endothelial cells was examined using monoclonal antibodies. Some rats were pretreated with indomethacin to rule out the effect of endogenous prostaglandin. The combined administration of lansoprazole and sofalcone brought about increased concentrations and immunoreactive areas of bFGF and a greater number of endothelial cells, compared with the ethanol-alone treatment. The number of myofibroblasts increased more significantly in the group treated with a combination of agents than in that treated with ethanol alone, ethanol plus sofalcone, or ethanol plus lansoprazole. Indomethacin pretreatment partly abolished the effects of single and combined administration of these agents. In conclusion, the mixed administration of lansoprazole and sofalcone accelerated the microvascular and connective tissue regeneration during the healing of ethanol-induced gastric mucosal damage. PMID- 9872518 TI - The possible role of vascular endothelial growth factor (VEGF) in gastric ulcer healing: effect of sofalcone on VEGF release in vitro. AB - Angiogenesis plays an important role in gastric ulcer repair. Several growth factors are involved in angiogenesis and, of these, vascular endothelial growth factor (VEGF) has received considerable attention because it is the only factor that acts specifically on endothelial cells and, unlike other angiogenic growth factors, has the hydrophobic signal peptide required for extracellular transport according to classical secretory pathways. We have lately demonstrated the role of VEGF in gastric ulcer repair. Previous reports confirmed that sofalcone has remarkable effects on gastric ulcer healing, which may be mediated by its stimulatory effect on prostaglandin (PG) release in gastric cells. Our data indicate that PGs stimulate VEGF expression in gastric fibroblasts. In this report we hypothesize that the clinical effect of sofalcone is mediated by VEGF expression and we demonstrate that sofalcone stimulates VEGF release by gastric fibroblasts in primary culture. PMID- 9872519 TI - Gastric urease activity is inversely associated with the success of treatment for Helicobacter pylori: effect of sofalcone. AB - Eradication therapy for Helicobacter pylori (H. pylori) has been established. However, the physiological factors influencing the success of treatment remain unclear. The aim of this study was to analyze these factors and to evaluate the efficacy of sofalcone on H. pylori eradication therapy. Forty-four H. pylori infected and peptic ulcer patients were enrolled in this study. Twenty-seven patients were treated with lansoprazole (LPZ, 30 mg o.d. for 1-8 weeks) and amoxicillin (AMPC, 500 mg q.i.d, 1-2 weeks), followed by 8 weeks of treatment with famotidine (FAM, 20 mg o.d.). Moreover, sofalcone (SOF, 100 mg t.i.d) was administered to 17 patients throughout the therapeutic period. Endoscopic and serologic evaluations and the urea breath test (UBT) were performed before therapy. At the endoscopic examination, mucosal samples were biopsied and then tissue myeloperoxidase (MPO) content, an index of neutrophil infiltration was measured. Cure of H. pylori infection was determined 8 weeks after the cessation of LPZ. This eradication regimen afforded an overall cure rate of 63.0% (17/27) without SOF and 76.5% (13/17) with SOF. In the control group, treatment success was inversely associated with pre-UBT value (gastric urease activity), whereas this association was not observed in the SOF group. Furthermore, in the patients exhibiting a high preUBT value (>40%), a twofold higher eradication rate was obtained by the administration of SOF. In patients who were successfully eradicated, mucosal MPO level was slightly higher than those of unsuccessful cases, whereas there was no significant association with serum pepsinogen (PG I, PG II) concentration and its ratio (PG I/PG II). These results suggest that a low UBT value is a factor predicting treatment success. SOF administration may improve the eradication rate, especially in the high-UBT subgroup. PMID- 9872520 TI - Preliminary study on a novel quadruple eradication therapy with a mucoprotective drug, sofalcone, for Helicobacter pylori infection. AB - To assess the effects of the mucoprotective drug sofalcone, which has direct and indirect effects on Helicobacter pylori in vitro, the eradication rate, adverse effects, and the quality of healing peptic ulcers were evaluated. Each study patient was given 500 mg t.i.d. amoxicillin and 200 mg t.i.d. clarithromycin. In addition, three different treatment regimens were compared: a standard dose (20 mg q.d.) of the proton pump inhibitor omeprazole (OAC), a double dose (20 mg b.i.d.) of omeprazole (Ox2AC), and a standard dose of omeprazole and a standard dose (100 mg t.i.d.) of sofalcone (OACS). Thirty-one H. pylori-positive patients were treated with OAC, 37 with Ox2AC, and 41 with OACS therapy. With an intention to-treat analysis, the eradication rates were 74.2% for OAC, 86.2% for Ox2AC, and 85.0% for OACS therapy. The incidence of side effects was 9.6% for patients given OAC therapy, 86.5% for Ox2AC, and only 7.5% for OACS-treated patients, which was significantly lower than the incidence in the Ox2AC group. High-quality peptic ulcer scars were observed after eradication therapy which included solfacone. Although it is necessary to conduct a randomized double-blind study to obtain definitive conclusions, our results indicate that this novel quadruple eradication therapy with solfacone is an efficacious regimen with a high eradication rate and positive effects on ulcer healing, combined with a low incidence of adverse events. PMID- 9872521 TI - Personalized education improves hepatitis B vaccination rate among physicians in Saudi Arabia. PMID- 9872522 TI - The infection control practices of general dental practitioners. PMID- 9872523 TI - Is filtered or mineral water good for us and our patients? PMID- 9872524 TI - Resolving the controversy on environmental cultures for Legionella: a modest proposal. PMID- 9872525 TI - More than 10 years of unrecognized nosocomial transmission of legionnaires' disease among transplant patients. AB - OBJECTIVE: To investigate a cluster of cases of legionnaires' disease among patients at a hospital. SETTING: A university hospital that is a regional transplant center. DESIGN: Retrospective review of microbiology and serology data from the hospital laboratories and prospective surveillance via the radiology department; a case-control study and environmental sampling within the hospital and from nearby cooling towers. RESULTS: Diagnosis of seven cases of legionnaires' disease in the first 9 months of 1996 led to recognition of a nosocomial outbreak that may have begun as early as 1979. Review of charts from 1987 through September 1996 identified 25 culture-confirmed cases of nosocomial or possibly nosocomial legionnaires' disease, including 18 in bone marrow and heart transplant patients. Twelve patients (48%) died. During the first 9 months of 1996, the attack rate was 6% among cardiac and bone marrow transplant patients. For cases that occurred before 1996, intubation was associated with increased risk for disease. High-dose corticosteroid medication was strongly associated with the risk for disease, but other immunosuppressive therapy or cancer chemotherapy was not. Several species and serogroups of Legionella were isolated from numerous sites in the hospital's potable water system. Six of seven available clinical isolates were identical and were indistinguishable from environmental isolates by pulsed-field gel electrophoresis. Initial infection control measures failed to interrupt nosocomial acquisition of infection. After extensive modifications to the water system, closely monitored repeated hyperchlorinations, and reduction of patient exposures to aerosols, transmission was interrupted. No cases have been identified since September 1996. CONCLUSIONS: Legionella can colonize hospital potable water systems for long periods of time, resulting in an ongoing risk for patients, especially those who are immunocompromised. In this hospital, nosocomial transmission possibly occurred for more than 17 years and was interrupted in 1996, after a sudden increase in incidence led to its recognition. Hospitals specializing in the care of immunocompromised patients (eg, transplant centers) should prioritize surveillance for cases of legionnaires' disease. Aggressive control measures can interrupt transmission of this disease successfully. PMID- 9872526 TI - A recurrent outbreak of nosocomial legionnaires' disease detected by urinary antigen testing: evidence for long-term colonization of a hospital plumbing system. AB - BACKGROUND: In 1994, a hospital reported an increase in nosocomial legionnaires' disease after implementing use of a rapid urinary antigen test for Legionella pneumophila serogroup 1 (Lp-1). This hospital was the site of a previous nosocomial legionnaires' disease outbreak during 1980 to 1982. METHODS: Infection control records were reviewed to compare rates of nosocomial pneumonia and the proportion of cases attributable to legionnaires' disease during the 1994 outbreak period with those during the same period in 1993. Water samples were collected for Legionella culture from the hospital's potable water system and cooling towers, and isolates were subtyped by monoclonal antibody (MAb) testing and arbitrarily primed polymerase chain reaction (AP-PCR). RESULTS: Nosocomial pneumonia rates were similar from April through October 1993 and April through October 1994: 5.9 and 6.6 per 1,000 admissions, respectively (rate ratio [RR], 1.1; P=.56); however, 3.2% of nosocomial pneumonias were diagnosed as legionnaires' disease in 1993, compared with 23.9% in 1994 (RR, 9.4; P<.001). In 1994, most legionnaires' disease cases were detected by the urinary antigen testing alone. MAb testing and AP-PCR demonstrated identical patterns among Lp-1 isolates recovered from a patient's respiratory secretions, the hospital potable water system, and stored potable water isolates from the 1980 to 1982 outbreak. CONCLUSIONS: There may have been persistent transmission of nosocomial legionnaires' disease at this hospital that went undiscovered for many years because there was no active surveillance for legionnaires' disease. Introduction of a rapid urinary antigen test improved case ascertainment. Legionella species can be established in colonized plumbing systems and may pose a risk for infection over prolonged periods. PMID- 9872527 TI - Controlling Legionella in hospital water systems: experience with the superheat and-flush method and copper-silver ionization. AB - OBJECTIVE: To evaluate the effect of copper-silver ionization on Legionella colonization and nosocomial legionnaires' disease and to compare the efficacy of metal ions versus the superheat-and-flush method of disinfection. DESIGN: Prospective determination over a 36-month period of copper and silver ion concentrations in the recirculating hot-water system, Legionella colonization of the hospital water distribution system, and cases of nosocomial legionnaires' disease. Retrospective comparison of results with the previous 13 years, during which the superheat-and-flush method was used. SETTING: The Pittsburgh Veterans' Affairs Health Care System (University Drive Division) acute-care hospital. INTERVENTION: Three copper-silver ionization systems were installed on the hot water distribution system in November 1994. RESULTS: The average number of cases of legionnaires' disease per year and the percentage of distal sites positive for Legionella pneumophila for the superheat-and-flush method versus the copper silver ionization method was six cases with 15% positivity versus two cases with 4% positivity, respectively. The reduction in Legionella colonization after copper-silver ionization was significant (P<.05) compared to the superheat and flush. Mean copper and silver ion concentrations (mg/L) were 0.29 and 0.054 from hot-water tanks, and 0.17 and 0.04 from distal outlets, respectively. CONCLUSIONS: We conclude that a properly maintained and monitored copper-silver ionization system was more effective than the superheat-and-flush method for reducing the recovery of Legionella from the hospital water distribution system. PMID- 9872528 TI - Tuberculosis, hepatitis B, rubella, rubeola, and varicella infection and immunity among medical school employees. AB - OBJECTIVE: To assess baseline health status of a medical school employee population and to assess this population's acceptance of vaccination and other interventions to reduce risk of disease transmission. DESIGN: A retrospective review of an employee health records database for a 4-year period. SETTING: A large, urban university hospital. PARTICIPANTS: 5,007 employees screened by employee health for immunity to vaccine-preventable illnesses and tuberculosis. RESULTS: 9.4% of the employees had positive tuberculin skin tests, with a conversion rate of 6.4% for those who had negative tests within the previous 2 years. Two individuals were identified who had active pulmonary tuberculosis. Fewer than 10% of the individuals for whom isoniazid chemoprophylaxis was recommended completed the 6 months of therapy. Most clinical employees (96.1%) did not have a history of prior hepatitis B virus (HBV) infection or immunization, but 77% of them subsequently completed the vaccination series. Most employees with a negative history for infection with or immunization against rubella, rubeola, and varicella had serological evidence of immunity (90.2%, 97.9%, and 87.2%, respectively). CONCLUSIONS: Review of aggregate employee health databases may assist individuals who must establish strategies for prevention of occupational illness and disease transmission in this specialized setting. While many employees at risk for HBV complete the vaccination series, strategies for improving this rate could be helpful. Substantial work is needed to analyze reasons why so few individuals for whom isoniazid chemoprophylaxis is recommended complete the therapy, and strategies tailored to the impediments identified should be implemented. PMID- 9872529 TI - Detection of Bordetella pertussis and respiratory synctial virus in air samples from hospital rooms. AB - OBJECTIVE: To evaluate the distribution of Bordetella pertussis and respiratory syncytial virus (RSV) in the hospital setting. DESIGN: Air samples were collected using filters in the hospital rooms of 12 children with pertussis and 27 children with RSV infection. Material eluted from these filters was subjected to RSV- and B pertussis-specific polymerase chain reaction (PCR) amplification. SETTING: Patients were hospitalized in private rooms in one of two referral centers, a university teaching hospital and a university-affiliated private children's hospital. PATIENTS: 12 children (16 days-3 years of age) with documented pertussis infection and 27 patients (10 days-7 years of age) with documented RSV infection. RESULTS: B pertussis DNA was detected in 7 (58%) of 12 rooms housing pertussis patients and in 16 (25%) of 63 total samples. B pertussis DNA was detected as far as 4 m away from the patient's bedside. The detection of B pertussis DNA in air samples did not change over the short duration of hospitalization. RSV RNA was detected in 17 (63%) of 27 rooms housing RSV infected patients and in 32 (22%) of 143 total samples. RSV RNA was detected at distances as far as 7 m from the patient's bedside and for up to 7 days of hospitalization. CONCLUSIONS: Using PCR-based detection methods, B pertussis DNA and RSV RNA both can be detected in air samples from the hospital rooms of infected patients. Both can be detected at large distances from a patient's bedside in a minority of cases. These detection methods are suitable for further studies of control measures used to contain nosocomial infections caused by both B pertussis and RSV. PMID- 9872530 TI - Epidemiological analysis defining concurrent outbreaks of Serratia marcescens and methicillin-resistant Staphylococcus aureus in a neonatal intensive-care unit. AB - OBJECTIVE: To describe the epidemiology, interventions, and molecular typing methods used during the investigation and control of concurrent outbreaks of Serratia marcescens and methicillin-resistant Staphylococcus aureus (MRSA) infections in a neonatal intensive-care unit (NICU). SETTING: A 206-bed women's and infants' hospital with a 48-bed NICU. DESIGN: A 22-week, prospective, descriptive study of all NICU infants with S marcescens or MRSA infection or colonization. Repetitive polymerase chain reaction (rep PCR) and pulsed-field gel electrophoresis (PFGE), respectively, were applied to the typing of S marcescens and MRSA isolates. INTERVENTIONS: Infants with S marcescens or MRSA infection or colonization were placed in isolation; all other infants were cohorted. A multidisciplinary task force implemented education for all hospital and medical staff regarding policies essential for outbreak control. Changes in physical setting and patient contact procedure were required to promote adherence to existing policies. RESULTS: Two premature infants had S marcescens infection, and five were colonized; rep PCR verified that both invasive and three of five colonizing isolates were related genotypically. Five bacteremic and 10 MRSA colonized infants were identified; PFGE confirmed that 12 of the isolates had similar electrophoretic patterns. S marcescens infection was eliminated from the NICU 3 weeks after interventions were initiated. MRSA infections also were eliminated, and MRSA colonization fell to below pre-outbreak rates within 8 weeks. Despite a 100% increase in NICU patient days per month during the subsequent 2 years, no further clusters of S marcescens or MRSA infection have occurred. CONCLUSIONS: Concurrent outbreaks of S marcescens and MRSA in an NICU were confirmed by genotyping of strains. Control was achieved by isolation and cohorting of patients and strict adherence to NICU policies and procedures. PMID- 9872531 TI - Dedifferentiation of the retinal pigment epithelium compared to the proliferative membranes of proliferative vitreoretinopathy. AB - PURPOSE: To examine the gene expression for melanogenesis of retinal pigment epithelial cells during dedifferentiation and to compare the condition to that of eyes obtaining anatomical success after surgery for proliferative vitreoretinopathy. METHODS: Gene expression for melanogenesis was determined by reverse transcriptase-polymerase chain reaction of tyrosinase and tyrosinase related protein-1 genes in normal and cultured retinal pigment epithelial cells and in proliferative membranes in patients with proliferative vitreoretinopathy. RESULTS: Gene expression for melanogenesis was classified into three types during dedifferentiation of retinal pigment epithelial cells: (1) tyrosinase-related protein-1 gene expression, (2) tyrosinase and tyrosinase-related protein-1 gene expression and (3) no expression of these genes. The expression of these genes were maintained better in mediums with basic fibroblast growth factor than in medium without basic fibroblast growth factor. Of the anatomically unsuccessful patients with proliferative vitreoretinopathy treated by surgery, 76.9% showed both tyrosinase and tyrosinase-related protein-1 gene expression; only 20% of the anatomically successful patients showed the gene expression. CONCLUSIONS: We reported three different conditions of retinal pigment epithelial cells based on gene expression for melanogenesis during dedifferentiation. The different condition of the retinal pigment epithelial cells may have some relationship to the anatomical results for proliferative vitreoretinopathy surgery. PMID- 9872532 TI - Activation of phospholipase C and guanylyl cyclase by endothelins in human trabecular meshwork cells. AB - PURPOSE: To characterize effects of endothelins on activities of phospholipase C (PLC) and nucleotide cyclases in human trabecular meshwork (TM) cells. METHODS: Cultured simian virus 40-transformed human TM (HTM-3) or non-transformed (HTM-16) cells were used. Changes in the PLC activity were determined by assaying the production of [3H] inositol phosphates. Accumulation of cyclic GMP or cyclic AMP in cell lysate was measured by radioimmunoassay. RESULTS: Endothelin-1 (ET-1; 1 microM) stimulated PLC in HTM-16 cells, but Sarafotoxin S6c (SRTX), an ET(B) receptor subtype-selective agonist (1 microM), did not. Similar results were obtained in HTM-3 cells: ET-1, but not ET-3 or SRTX, activated PLC in a dose dependent manner, with a calculated EC50 of 646 pM. The peptide also stimulated the accumulation of cGMP in a concentration-dependent manner with an EC50 of 37.2 pM. ET-3 or SRTX was not effective except at much higher concentrations. Both the PLC and guanylyl cyclase stimulation induced by ET-1 (10 nM) were completely inhibited by pretreating the cells with BQ-123 (<10 microM), an ET(A) receptor selective antagonist, but not by BQ-788 (10 microM), an ET(B) receptor subtype specific antagonist. Neither ET-1 nor ET-3 stimulated adenylyl cyclase activity in HTM-3 cells at concentration as high as 1 microM. CONCLUSION: ET-1 activates PLC and guanylyl cyclase in TM cells. Potency profiles of ET receptor agonists and antagonists suggest that the ET(A) receptor subtype is involved in both actions of ET-1. The effects of the ET peptides in TM cells are interesting and could be part of the mechanism of their IOP-lowering effect. PMID- 9872533 TI - Finger-like projections of plasma membrane in the most senescent fiber cells of human lenses. AB - PURPOSE: To corroborate the findings of finger-like membrane projections in monkey and baboon lenses, in human lens fiber cells. METHODS: Normal human lenses, two months to 76 years old, as well as age-related nuclear cataracts, were immersion fixed in 2% paraformaldehyde-0.2% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.2 for 24 h at room temperature, cut into 200-500 microm thick sections along the equatorial axis, fixed for an additional 12-18 h at room temperature, dehydrated in an ascending ethanol series and embedded in Lowicryl K4M. Semi-thick sections (0.25-1.0 microm) were cut, absorbed onto 75 or 100 nickel slotted grids, labeled with anti-MIP 26 or phalloidin, stained with 2% uranyl acetate and viewed by transmission electron microscopy at 100 kV. RESULTS: Transmission electron microscopy micrographs demonstrated the presence of finger like plasma membrane projections measuring 0.16-0.25 microm in diameter and 1.0 6.5 microm in length with bulbous terminal tips in the most senescent fiber cells in two-month and older normal lenses, as well as, in nuclear cataracts. These projections appeared to overlie furrowed membrane domains in the extracellular space, as well as project into the cytosol along the cytosolic leaflet of plasma membrane. CONCLUSIONS: The results extend the findings in monkey and baboon lenses, to the human lens, and demonstrate that these projections, which sparsely label with antiserum against MIP 26, but not filamentous actin, not only extend into the extracellular space, but also project inward into the cytosol. PMID- 9872534 TI - Early changes in iris blood vessels in exfoliation syndrome. AB - PURPOSE: To look for possible changes in iris blood vessels that might precede the formation of true exfoliation material, and to correlate these changes, if present, to aging of the iris. METHODS: Iris blood vessels of 10 clinically normal irises from 10 eyes (ages: 1 day-80 years), 4 irises with exfoliation (ages: 64-80 years) and 3 irises from primary open-angle glaucoma (POAG) (ages: 27-65 years) were examined with an electron microscope. RESULTS: Pronounced multilayering of the basal lamina around the vessels, abnormal excessive formation of microfibrils, presence of atrophic invaginations in the outer cell membranes of endothelial cells containing interlacing basal lamina, and an apparent decrease in the perivascular collagen fibers were evident in exfoliative iris vessels devoid of true exfoliation material. Similar, but much milder, changes were evident in normal aged and POAG irises. CONCLUSIONS: Abnormal elastic tissue and/or basal lamina production plays a role early in the formation of exfoliation material around iris blood vessels. PMID- 9872535 TI - Human primary Sjogren's syndrome autoantibodies as mediators of nitric oxide release coupled to lacrimal gland muscarinic acetylcholine receptors. AB - IgG obtained from sera of primary Sjogren's syndrome (pSS-IgG) patients and its interaction with M3 muscarinic cholinoceptors of rat exorbital lacrimal glands were studied by indirect immunofluorescence (IFI) and binding assay. Primary Sjogren's syndrome IgG stained epithelial cells with a continuous fluorescence pattern. The IFI imagen was attenuated by incubating the pSS-IgG with a synthetic peptide corresponding to the second extracellular loop of M3 muscarinic cholinoceptor. Primary SS-IgG was also able to bound irreversibly to muscarinic acetylcholine receptors (mAChRs) displacing the specific cholinergic antagonist QNB. Moreover, these antibodies triggered intracellular signals coupled to M3 muscaric cholinoceptors such as nitric oxide synthase (NOS) activation and cGMP production. Both primary Sjogren's syndrome IgG effects mimicked carbachol action and were abrogated by specific muscarinic antagonist 4-DAMP. The nitric oxide pathway through muscarinic cholinoceptors activation by pSS-IgG on rat exorbital lacrimal gland is also described. We proposed that chronic interaction of these autoantibodies on lacrimal gland muscarinic acetylcholine receptors could lead to tissue damage through nitric oxide release after immunological stimulation. PMID- 9872537 TI - Fourth module of Xenopus interphotoreceptor retinoid-binding protein: activity in retinoid transfer between the retinal pigment epithelium and rod photoreceptors. AB - PURPOSE: Interphotoreceptor retinoid-binding protein (IRBP), an extracellular protein believed to support the exchange of retinoids between the neural retina and retinal pigment epithelium (RPE) in the vertebrate eye, exhibits a modular, i.e., repeat, structure. The present study was undertaken to determine whether an individual module of IRBP has activity in retinoid transfer between the RPE and rod photoreceptors. METHODS: The retinoid transfer activity of a recombinant protein corresponding to the fourth module of Xenopus laevis IRBP (X4IRBP) was examined in two ways. First, X4IRBP was tested for its ability to support the regeneration of porphyropsin in detached/reattached Xenopus retina/RPE-eyecups. Following illumination and removal of native IRBP, Xenopus eyecups supplemented with 42 microM X4IRBP or (as a control) Ringer's solution were incubated in darkness and then analyzed for regenerated porphyropsin. Second, toad (Bufo marinus) RPE-eyecup preparations were used to evaluate X4IRBP's ability to promote the release of 11-cis retinal from the RPE. RESULTS: The regeneration of porphyropsin in X4IRBP-supplemented Xenopus retina/RPE-eyecups (0.45 +/- 0.04 nmol; mean +/- SEM, n = 11) exceeded that in controls (0.13 +/- 0.02 nmol, n = 11). For promoting the release of 11-cis retinal from the toad RPE, 42 microM X4IRBP was more effective than equimolar bovine serum albumin although considerably less than that of 26 microM native bovine IRBP. CONCLUSIONS: The results indicate a low but significant activity of IRBP's fourth module in reactions relevant to retinoid exchange. PMID- 9872536 TI - Effect of substance P, insulin-like growth factor-1 and vasoactive intestinal polypeptide on corneal re-epithelialization in galactosemic rats. AB - PURPOSE: The purpose of the study was to examine the effect of topical application of substance P (SP), insulin-like growth factor-1 (IGF-1) and vasoactive intestinal polypeptide (VIP) on corneal re-epithelialization in galactosemic rats. METHODS: Experimental galactosemia was induced by feeding a diet containing 30% galactose for 4-6 months. The corneal epithelium was debrided bi-laterally by scraping with a blunted scalpel blade. One eye (control) received only a saline solution whilst the other eye received a solution of SP and/or IGF 1 or VIP. A single drop of control or test solution was administered 4 times daily until wound closure. Corneas were stained with fluorescein and videotaped under ultraviolet illumination at regular time intervals after debridement. After digitizing the video image, the wound area was calculated using an image analysis program (NIH Image). RESULTS: Corneal re-epithelialization was significantly delayed in galactosemic compared to normal animals. Rates of healing were 1.3 +/- 0.07 mm2/hour and 1.02 +/- 0.02 mm2/hour for normal and galactosemic animals, respectively. Topical application of SP in concentrations ranging from 25 pg/ml up to 250 microg/ml had no significant effect on the rate of corneal re epithelialization. Similarly, IGF-1 (1 microg/ml) or VIP (1 microg/ml) when applied alone did not affect re-epithelialization. Furthermore, resurfacing of the debrided area was not affected by co-application of SP (250 microg/ml) and IGF-1 or VIP. CONCLUSIONS: Independent or combined topical application of SP, VIP or IGF-1 at the concentrations tested, does not modulate corneal epithelial wound healing in rats with galactosemia induced by 30% galactose. PMID- 9872538 TI - Surgical treatment of biliary atresia in the liver transplantation era. AB - Biliary atresia (BA) still remains one of the most intractable gastrointestinal diseases in infancy despite the concerted efforts of pediatric surgeons all over the world. The introduction of liver transplantation has revolutionized the protocols for the treatment of this condition. In this editorial, the role of hepatic portoenterostomy (the Kasai procedure) in the surgical treatment of BA in the "transplantation era" will be discussed. PMID- 9872539 TI - Monitoring carcinoembryonic antigen in colorectal cancer: is it still useful? AB - The results of a study conducted to determine the usefulness of carcinoembryonic antigen (CEA) monitoring in the follow-up of patients with resected colorectal cancer are reported herein. The subjects of this study were 125 patients in whom CEA had been determined preoperatively and 239 patients in whom CEA had been monitored postoperatively. The results revealed increased preoperative CEA in only 24% of the subjects, and that this increment was correlated with subsequent more advanced tumor stage and a higher recurrence rate (P < 0.01). The postoperative CEA level exceeded the threshold in 71% of the patients affected by recurrence, 94.4% of whom developed liver metastases and 50%, nonhepatic recurrence. This marker showed elevated sensitivity for liver metastases (99%), whereas the sensitivity was lower for nonhepatic recurrence of the disease (94%). Thus, we concluded that CEA monitoring can be useful for preoperative colorectal tumor grading, even if its validity in the early diagnosis of recurrence is problematic, especially in terms of radical repeated surgery and survival. PMID- 9872540 TI - Additional techniques adopted for major vascular anastomoses during orthotopic liver transplantation. AB - Of 372 patients who underwent liver transplants between January 1985 and March 1995, 7 required variations in vascular anastomoses due to discrepancies in the size-match of the donor and recipient vessels, the presence of a thrombosed hepatic artery or portal vein, or complete absence of the portal vein. The techniques described herein enabled us to perform successful transplantation in all patients. Although rethrombosis of the portal vein developed in one patient after a third transplant, this patient remains clinically well. The use of an operating microscope and the harvesting of an extended length of donor superior mesenteric vein or vascular grafts of the donor iliac or saphenous vessels, for potentially difficult transplants, are invaluable techniques. PMID- 9872541 TI - The long-term survival rates of patients after repair of abdominal aortic aneurysms. AB - This study was undertaken to examine the long-term survival rates of patients following abdominal aortic aneurysm (AAA) repair in comparison with an age matched normal population, and to determine by multivariate analysis the factors influencing long-term survival. Of 125 patients who underwent AAA repair prior to July 1986, 13 died during hospitalization. Of these 13 patients, 6 who suffered aneurysmal rupture all died within 30 days. The survival rate of patients with ruptured aortic aneurysms was significantly lower than that of those with nonruptured aneurysms. Of the 112 patients surviving hospitalization, 85 died within 0.48 to 24 years after their operation. The long-term survival rate of patients who had suffered a preoperative cardiovascular event was significantly lower than that of those who had not suffered a preoperative cardiovascular event. The actual survival rate was significantly lower than the expected survival rate. According to a multivariate analysis, the significant predictors of late survival were age, aneurysmal rupture, and chronic renal failure in all the patients, and age, chronic renal failure, and pre- and postoperative cardiovascular events in patients who did not die in hospital. These findings indicate the importance of improving immediate perioperative management of ruptured AAA and that cardiovascular events should be prevented, or treated during long-term follow-up. PMID- 9872542 TI - The therapeutic effect of hypertonic solutions on the changes in the effective circulating plasma volume in acute necrotizing pancreatitis in rats. AB - While hypovolemia or hypovolemic shock is dominant in the early stage of severe acute pancreatitis, there have been few studies on the effects of hypertonic solutions in the management of this disease. We conducted this study to evaluate the therapeutic effects of hypertonic saline solutions (HS) on the course of severe acute pancreatitis in rats. Pancreatitis was induced in male Wistar rats by injecting a 5% solution of sodium taurocholate into the biliopancreatic duct. The effective circulating plasma volume (ECPV) was measured using radioiodinated [125I]bovine serum albumin. Samples of blood and of ascitic fluid were obtained 3, 6, and 12 h after the onset of pancreatitis. Lactated Ringer's solution (LR) and HS were administered consecutively for 3 h beginning 3 h after the induction of pancreatitis. ECPV was measured 6 h after the onset of pancreatitis. The survival rates were investigated for up to 10 days. The mean ECPV decreased significantly from 24.9 +/- 1.1 ml/kg before disease onset to 11.5 +/- 1.3 ml/kg 6 h postoperatively. LR failed to achieve a normal value for ECPV even following a 150 ml/kg infusion. HS200 and HS300 restored the ECPV to the normal level, and with smaller volumes infused. All rats in the untreated group died within 3 days. LR and HS improved the survival rates, with the infusion of HS200, 100 ml/kg, thus attaining a 45% survival at 10 days. PMID- 9872543 TI - Abdominal incisional hernia occurring after an esophagectomy reconstructed with a gastric tube through an antesternal route: report of a case. AB - A Japanese man, who had undergone a subtotal esophagectomy reconstructed with a gastric tube through an antesternal route for esophageal carcinoma 16 years previously, was admitted to our hospital because of an abdominal incisional hernia. The abdominal incisional hernia was in his upper abdomen and was difficult to push back into the intraabdominal cavity by hand. The hernia was successfully repaired by operation. We thus conclude that an abdominal incisional hernia is a rare but important late-phase complication occurring after an esophagectomy reconstructed with either an antesternal or retrosternal route, and an operation should be the treatment of choice. PMID- 9872544 TI - Major liver resection for non-Hodgkin's lymphoma in an HIV-positive patient: report of a case. AB - A very unusual clinical presentation of non-Hodgkin's lymphoma (NHL) of the liver is reported herein. The patient was a 35-year-old male who had been HIV-positive since 1987. Following an episode of acute pain in the right upper abdominal quadrant, ultrasonography (US) and computed tomography (CT) were performed, revealing a nodular lesion, 2.5 cm in diameter, localized in the eighth segment of the liver. Despite the fact that the lesion became significantly enlarged over a 6-month period, three repeated percutaneous biopsies proved negative. Finally, his increasing pain and the lack of a definitive diagnosis prompted us to perform a right hepatectomy. The patient had an uneventful postoperative course and is well 1 year after his operation. Establishing a diagnosis of extranodal lymphoma can be difficult, especially in HIV-positive or AIDS patients. Thus, performing a laparotomy is justified to confirm a diagnosis and decide upon the most appropriate treatment. However, in about 10% of these patients, only surgical resection allows for the diagnosis. In accordance with other reports, our experience confirms that, in contrast with AIDS patients, HIV-positive patients have a similar prognosis as non-HIV patients, and are suitable candidates for even major surgical procedures. PMID- 9872545 TI - Pancreatic mucin-producing adenocarcinoma associated with a pancreatic stone: report of a case. AB - We present an autopsy case of an 83-year-old Japanese man with a mucin-producing adenocarcinoma accompanied by pancreatolithiasis in the head of the pancreas. He suffered from obstructive jaundice and died of disseminated intravascular coagulation. He did not normally drink alcohol and had no history of chronic pancreatitis. The autopsy findings revealed a mucinous cystic tumor, composed of multiple dilatated branches, in the head of the pancreas. Histological examinations showed papillary adenocarcinoma, which scirrhously infiltrated the distal common bile duct with perineural invasion and lymph node involvement. He was thus diagnosed to have mucin-producing branch-type cancer in the head of the pancreas. The main pancreatic duct was dilated, and the residual pancreatic tissue showed moderate fibrosis and parenchymal atrophy. A stone was observed in a dilated branch of the primary lesion. To the best of our knowledge, there have only been five previously reported cases of mucin-producing tumor associated with pancreatolithiasis. Intraductal calcification is a major characteristic of chronic pancreatitis, but it is clinically important not to misdiagnose cancers associated with pancreatolithiasis such as chronic pancreatitis. PMID- 9872546 TI - Obstructive Ileus caused by blood clot after emergency total gastrectomy in a patient with hemophilia A: report of a case. AB - We report herein the extremely unusual case of a patient with hemophilia A who developed a hematoma ileus soon after undergoing an emergency total gastrectomy. Postoperatively, the patient was found to have an obstruction in the small intestine that was difficult to differentiate from intussusception. Reoperation revealed the cause of the obstruction to be a large blood clot. This case report serves to demonstrate that careful hemostasis and observation during surgery is essential in patients with hemophilia. PMID- 9872547 TI - Development of an ulcer in the side-to-side anastomosis of a jejunal pouch after proximal gastrectomy reconstructed by jejunal interposition: report of a case. AB - We present herein the first reported case of a deep ulceration developing in the pouch of a jejunal side-to-side anastomosis following the interposition of a jejunal U-shaped pouch (jejunal pouch), performed as reconstruction. A 47-year old woman underwent a proximal gastrectomy and interposition of a jejunal pouch for early gastric cancer, 4 months after which she began to develop a fever. Esophagogastroscopy revealed a deep ulceration in the side-to-side anastomosis of the jejunal pouch. Her serum gastrin level was much higher than the normal range, and 24-h monitoring of the intraremnant stomach pH revealed that it was below 7.0 at night. A peptic ulcer was thought to be one of the causes of the pouchitis. Due to the drug-resistant ulceration and fever, a reoperation was performed, in which the jejunal pouch and remnant stomach were removed and Roux-en-Y reconstruction was done. There were no postoperative complications, and the fever resolved after surgery. Reconstruction of the jejunal pouch after proximal gastrectomy is recommended from the standpoint of quality of life (QOL); however, further studies on the size of the remnant stomach and the length of the interposed jejunal pouch are necessary. PMID- 9872548 TI - Postoperative management following massive hepatectomy in a patient with Dubin Johnson syndrome: report of a case. AB - We report herein the case of a 46-year-old man with Dubin-Johnson syndrome (DJS) who was referred to our hospital to undergo a right hepatic lobectomy for hepatocellular carcinoma. His complicated postoperative conjugated hyperbilirubinemia was successfully treated by hemopurification based on the increased level of serum hepatocyte growth factor (HGF). It is considered that hemopurification based on the early postoperative HGF levels has beneficial effects for patients with DJS; however, the specific role of hemopurification in the conjugated hyperbilirubinemia that develops postoperatively in these patients has not been determined. PMID- 9872550 TI - Demonstration of microcystic disease of the pancreas by magnetic resonance cholangiopancreatography: report of a case. AB - A case of microcystic disease of the pancreas which was clearly demonstrated by magnetic resonance cholangiopancreatography (MRCP) is reported herein. Cystic dilatation of the pancreatic duct was recognized by computed tomography scanning and endoscopic retrograde cholangiopancreatography (ERCP). Furthermore, the existence of microcystic clusters surrounding the dilated pancreatic duct were clearly visualized by MRCP. These microcystic clusters were strongly suspected preoperatively of having caused dilatation of the major pancreatic duct. Based on these findings, a distal pancreatectomy was performed. The operative specimen showed no accumulation of mucin and no evident lesions in the dilated pancreatic duct, being inconsistent with the entity of a mucus-producing tumor. Pathological examination revealed that the inner parts of microcysts constituted columnar epithelium with mucus production and papillary growth. Thus, a final histological diagnosis of intraductal papillary adenoma with idiopathic pancreatic duct ectasia was confirmed. In conclusion, MRCP, being a less aggressive diagnostic procedure than ERCP, proved extremely useful for obtaining precise information on cystic lesions of the pancreas in this patient. PMID- 9872549 TI - Calcitonin-producing pancreatic somatostatinoma: report of a case. AB - A 59-year-old woman was hospitalized due to a 1-year history of diarrhea and weight loss. Echography and computed tomography of the abdomen revealed a 10 x 7 cm solid mass in the tail of the pancreas and gallstones, while selective celiac angiography revealed the presence of a hypervascular mass. High levels of somatostatin and calcitonin were detected in the plasma, 70 pg/ml (normal range <28 pg/ml) and 5550 pg/ml (normal range 37 +/- 8 pg/ml), respectively. This tumor was thus removed by means of a distal pancreatectomy and a splenectomy. After the pancreatic tumor was removed, the elevated levels of plasma somatostatin and calcitonin returned to the normal ranges, and the persistent diarrhea also dramatically disappeared. A postoperative immunohistochemical study showed the tumor cells to be diffusely positive for somatostatin and calcitonin. These results clearly indicate this patient to be a case of calcitonin-producing pancreatic somatostatinoma. PMID- 9872551 TI - Hypoproteinemia caused by tricuspid regurgitation: report of a case. AB - A case of tricuspid regurgitation (TR) complicated by severe hypoproteinemia is presented herein. A 68-year-old man who had undergone coronary artery bypass grafting (CABG) for postinfarction angina suffered repeated inferior myocardial infarction due to obstruction of the proximal right coronary artery, 3 years after which he developed systemic edema. Investigations revealed TR associated with hypoproteinemia; however, treatment consisting of aggressive diuretic therapy and albumin administration proved ineffective. The hypoproteinemia manifested as protein-losing enteropathy clinically, and the tricuspid valve was replaced to eliminate high venous pressure. The serum protein levels became normalized after the operation. Although TR is generally well tolerated in the absence of pulmonary hypertension, surgical management is recommended for patients with severe protein deficiency resistant to medical treatment. PMID- 9872552 TI - Ligation of medically refracted patent ductus arteriosus (PDA) in an extremely low body weight premature infant. AB - Medically refracted patent ductus arteriosus (PDA) in an extremely low birth weight (ELBW) preterm (gestation 24 weeks 2 days) infant was successfully ligated under general anesthesia in the neonatal intensive care unit (NICU). Pharmacological agents are more effective to close PDA in preterm infants than in full-term infants, although within 48 h three doses of indomethacin were not sufficient to close PDA in this case. At the age of 69 h the infant developed severe symptoms including bradycardia, systemic hypotension, pulmonary hypertension, diastolic steal (reverse distal aorta flow velocity), and anuria. A PDA ligation was thus performed surgically at 72 h of age. General anesthesia and surgical stress were tolerated by this 531 g infant. Postoperatively all symptoms improved dramatically and the general conditions were stable. On the 38th day the endotracheal tube was extubated and on the 50th day nasogastric milk feeding was started. The oxygen supply was weaned on the 78th day. Growth and development until 6 months were within the normal range of very low birth weight infants. A surgical ligation as early as possible in medically refracted PDA in an ELBW infant is thus considered to be a safe and effective treatment. It prevents the development of further complications of cardiopulmonary vascular problems. Color Doppler echocardiography can reliably measure the PDA size, flow velocity, and hemodynamic changes of persistent PDA, even in tiny infants. PMID- 9872553 TI - Occlusion of the abdominal aorta caused by enlargement of the false lumen after graft replacement for a DeBakey type IIIb dissecting aneurysm: report of a case. AB - We report herein a case of abdominal aorta occlusion caused by enlargement of the false lumen after a graft replacement. The patient was a 70-year-old man who underwent a graft replacement of the descending thoracic aorta for a DeBakey type IIIb dissecting aneurysm. Digital subtraction angiography performed on postoperative day 18 revealed an abdominal aorta occlusion caused by enlargement of the false lumen. A new intimal defect was found in the aortic wall 2 cm distal to the suture line, the cause of which was suspected to be clamp injury during the initial operation. Graft replacement of the abdominal aorta was subsequently carried out. Postoperatively, he had no complications, and digital subtraction angiography showed excellent reconstruction of the abdominal aorta. The patient was discharged from hospital 1 month after his second operation. PMID- 9872554 TI - Idiopathic renal arteriovenous fistula demonstrating a huge aneurysm with a high risk of rupture: report of a case. AB - This paper reports a case of an idiopathic renal arteriovenous fistula demonstrating a huge guitar-shaped aneurysm, which required a total nephrectomy. Although the patient insisted on having been asymptomatic, echocardiography and cardiac catheterization clearly revealed that the influence on hemodynamics was unexpectedly significant. Interestingly, the aneurysm was also considered to have a high risk of rupture, which was preoperatively suggested by an aneurysm wall projection discernible on both computerized tomograms and angiograms, and this suspicion was also convincingly supported by the perioperative and histopathological findings. PMID- 9872555 TI - Multiple myeloma mimicking bone metastasis from breast cancer: report of a case. AB - A 72-year-old woman with a history of early breast cancer suffered a fracture of the eighth thoracic vertebra resulting in paraplegia. Magnetic resonance imaging (MRI) showed spinal cord compression by a tumor between the ninth and tenth thoracic vertebrae. Local radiotherapy was begun under the diagnosis of metastatic breast cancer, but bone marrow aspiration and biopsy subsequently revealed plasma-cell proliferation rather than adenocarcinoma. This case report serves to demonstrate that clinicians should consider multiple myeloma as a cause of lytic bone lesions without extraskeletal metastases even in patients with a history of breast cancer. PMID- 9872556 TI - Subcarinal neurogenic tumor: report of a case. AB - We present herein the rare case of a 63-year-old man in whom a subcarinal tumor, demonstrated by enhanced chest computed tomograms (CT), was subsequently confirmed to be a neurilemmoma by histological examination following tumor resection through a diagnostic thoracoscopy. Magnetic resonance imaging (MRI) and transesophageal ultrasonogram findings excluded the possibility of malignant lymphoadenopathy. As the patient was also found to have an elevated level of the squamous cell carcinoma (SCC) tumor marker which did not resolve postoperatively, close follow-up will be required. PMID- 9872557 TI - Intrapulmonary cystic lymphangioma: report of a case. AB - We describe herein the case of a 52-year-old man found to have a solitary pulmonary lymphangioma. Computed tomography (CT) scan demonstrated a nodule with homogeneous density and a well-defined border in the lower lobe of the right lung. T2-Weighted magnetic resonance imaging (MRI) revealed a nodule with homogeneously high signal intensity. The tumor was removed by partial resection, and pathologic examination confirmed a diagnosis of primary lymphangioma of the lung. PMID- 9872558 TI - Primary bronchopulmonary fibrosarcoma: report of a case. AB - We herein report a case of primary bronchopulmonary fibrosarcoma in a 70-year-old man. The patient was referred to our hospital for investigation of hemosputum and an abnormal shadow. On admission, chest radiograph and computed tomography scan showed a mass lesion in right S3 and an infiltrative shadow in the right upper lobe. Transbronchial biopsy specimens showed findings of malignancy, and adenocarcinoma was suspected. A right pneumonectomy was performed, and pathologic examination confirmed a diagnosis of fibrosarcoma. The patient had an uneventful recovery and no sign of recurrence has been found in the year since his operation, although strict follow-up is essential. PMID- 9872559 TI - Pulmonary histoplasmosis in a Japanese male: report of a case. AB - A case of pulmonary histoplasmosis, which is rare in Japan, is reported herein. A 43-year-old man who had worked in Mexico for 2 years and had come back to Japan 3 months earlier, presented at our hospital because of an abnormal shadow on his chest roentogenogram with no symptoms. His chest roentogenogram as well as chest computed tomograms revealed a 2-cm-diameter nodule in the anterior basal segment of his right lung and an enlargement of the subcarinal lymph node. Although these pictures seemed to indicate an advanced lung cancer, no malignant cells were found based on the brushing cytology findings after bronchoscopy. An exploratory thoracoscopic tumor resection and biopsy of the enlarged lymph node led to a histological diagnosis of an abscess due to histoplasma. The hospital course was uneventful. Postoperatively, amphotericin B was administered for 1 year. This is the eighth case of pulmonary histoplasmosis reported in Japan. A pulmonary nodule together with mediastinal lymphoadenopathy seems to be characteristic in these patients. Histoplasmosis should therefore be considered in the differential diagnosis, since overseas travel has now become common-place for the Japanese. PMID- 9872561 TI - Two cases of synchronous multiple thymoma. AB - We report two cases of synchronous double primary thymoma without myasthenia gravis. These cases suggest the possibility of multicentric thymoma and confirm the validity of a complete thymectomy. PMID- 9872560 TI - Adenocarcinoma arising in a tailgut cyst: report of a case. AB - We report herein the unusual case of a 66-year-old woman found to have adenocarcinoma arising in a tailgut cyst. The patient had been observed for 6 months following the discovery of a presacral cystic mass measuring 10 x 9 cm for which she had refused surgery. The serum tumor marker, carcinoembryonic antigen, became slightly elevated, and diagnostic imaging distinctly revealed a tumorous lesion with papillary projection into the cyst lumen. The cystic mass was then excised through the transsacral approach. The pathological findings were compatible with moderately differentiated adenocarcinoma arising in a tailgut cyst. This entity is extremely rare, and only six cases, including our own, have been reported in the English literature. Early complete excision is advised because it is almost impossible to determine for certain whether presacral cystic masses are benign or malignant prior to surgery. PMID- 9872562 TI - Development of primary leiomyosarcoma of the sternum postirradiation: report of a case. AB - We report herein the case of a 30-year-old man who developed a primary leiomyosarcoma (LMS) 11 years after undergoing a median sternotomy for mediastinal seminoma followed by 50 Gy radiotherapy. He was given two courses of chemotherapy, resulting in 90% tumor regression, after which resection of the tumor with adjacent chest wall structures was carried out. Reconstruction was performed using a methylmethacrylate prosthesis prepared preoperatively. Postoperatively, he received two additional courses of chemotherapy and has had no sign of recurrence for 45 months. PMID- 9872563 TI - Cardiopulmonary bypass established via a sutured vascular graft in the presence of extraanatomic bypass. AB - Cardiopulmonary bypass (CPB) was established via a sutured collagen sealed knitted Dacron graft in two patients who had undergone extraanatomic bypasses for lower limb ischemia, whose ascending aorta was not suitable for cannulation. One patient, with a history of femorofemoral bypass, underwent surgery for a ruptured aortic arch aneurysm (AAA) and the other patient, with a history of axillobifemoral bypass for chronic dissection involving the aortic arch and descending aorta, had unstable angina and underwent coronary artery bypass grafting. This technique of perfusion was found to be safe and effective for patients with an impaired ascending aorta who have undergone an extraanatomic bypass for the lower limb. PMID- 9872564 TI - Regulation of ion content in primary cultures from reabsorptive ducts of human sweat glands studied by X-ray microanalysis. AB - X-ray microanalysis was used to investigate whether cAMP- and/or Ca2+-activated regulation of chloride and potassium efflux is expressed in primary cultures of sweat gland duct cells. The effects of extracellular UTP and ATP on the duct cells, and the signalling system involved in the response to ATP was also studied. Primary cultures from duct cells of human sweat glands responded to 1 microM carbachol, 2 microM of the Ca2+ ionophore A23187, or 5 mM 8-bromo-cAMP stimulation for 5 min, resulting in a decrease in cellular Cl and K concentrations. 50 microM 5-nitro-2-(3-phenylpropyl-amino)-benzoic acid (NPPB), a Cl- channel blocker, can inhibit the decrease in Cl concentration induced by cAMP. Extracellular (200 microM) ATP caused a decrease of Cl and K in cultured duct cells, while (200 microM and 2 mM) UTP was ineffective. Both the phosphoinositidase C inhibitor U73122 (10 microM) and the absence of extracellular Ca2+ abolished the ATP-induced decrease in Cl and K content. Alloxan (1.25 mM), an adenylate cyclase inhibitor, had an inhibitory effect on the response to ATP. The decrease in K, but not in Cl, content in the cells elicited by ATP was blocked by prior incubation with 100 ng/ml pertussis toxin, indicating the coupling of ATP to pertussis toxin-sensitive G-proteins. In conclusion, both Ca2+- and cAMP-dependent Cl- permeability is present in primary cultures from duct cells of human sweat gland. The response to ATP can be mediated both by Ca2+- and by cAMP-dependent pathways, and is coupled to pertussis toxin-sensitive G-proteins. PMID- 9872565 TI - In vitro bovine ciliary body/epithelium in a small continuously perfused Ussing type chamber. AB - Our goal is to assess the viability of an in vitro preparation of bovine ciliary body/epithelium (CBE) in a small volume Ussing-type chamber. A new small volume Ussing-type chamber with continuous perfusion was developed for bovine CBE. The trans-CBE electrical parameters were monitored and the electrical responses of the CBE to ouabain (1 and 0.01 mM) were recorded. The trans-CBE fluxes of [14C]-L ascorbate and [3H]-L-glucose were also studied. The bovine CBE preparation was stable inside the chamber in terms of its potential difference (PD), short circuit current (SCC) and trans-CBE resistance. They were -0.51+/-0.05 mV (aqueous side negative), -5.43+/-0.04 microAcm-2 and 94+/-2 Q.cm2 (mean s.e.m., n=35), respectively. The preparation hyperpolarised when 0.01 mM ouabain was administered to the aqueous side, depolarised when ouabain was applied to the stromal side. [3H]-L-glucose diffusion was about 74 nEq h(-1)cm(-2) in either direction (n=12). Taking the area magnification factor of the CBE into consideration, the diffusional L-glucose flux across the bovine CBE was comparable to other tight epithelia. A significant net ascorbate flux (0.26+/ 0.05 nEq h(-1)cm(-2), n=4, p<0.01) was found in the stroma to aqueous direction. We have developed a viable in vitro bovine CBE preparation which was (1) electrically stable, (2) responsive to ouabain, (3) tight to L-glucose diffusion, and (4) capable of actively secreting ascorbate. A net trans-CBE chloride transport (0.81+/-0.30 microEq h(-1)cm(-2), n=12, p=0.01) from stromal to aqueous side was found in the present in vitro model under short-circuited conditions. PMID- 9872566 TI - Rapid adhesion and spread of non-adherent colon cancer Colo201 cells induced by the protein kinase inhibitors, K252a and KT5720 and suppression of the adhesion by the immunosuppressants FK506 and cyclosporin A. AB - We examined alterations in cell morphology and expression of adhesion molecules in response to a general protein kinase inhibitor K252a treatment of non-adherent colon adenocarcinoma Colo201 cells. K252a induced rapid cell adhesion and spreading with concomitant formation of actin stress fibers. A protein kinase A inhibitor KT5720 also induced cell adhesion, but the rate of spread was slower than that seen with K252a. These adhesions were mediated by integrin molecules since cell adhesion required Mg2+, Mn2+ or Ca2+, and was inhibited by monoclonal antibodies for integrins alpha2 and beta1. Indirect immunofluorescence microscopic observations revealed that integrin alpha2 and beta1 molecules in K252a-treated cells were concentrated at sites of focal adhesion, but expressions of integrin molecules were not modulated. Tyrosine phosphorylation of focal adhesion kinase (FAK) and paxillin increased during K252a- or KT5720-induced cell adhesion. Immunosuppressants FK506 and cyclosporin A suppressed the K252a-induced cell adhesion and abolished tyrosine phosphorylation of cellular proteins including FAK and paxillin. Furthermore, W7 and calmidazolium, inhibitors of calmodulin, also inhibited the cell adhesion. Based on findings that FK506 and cyclosporin A are inhibitors of the calcium calmodulin-dependent protein phosphatase, calcineurin, this phosphatase may regulate integrin-dependent cell adhesion and spread of Colo201 cells. This Colo201 cell model provides a pertinent system for studying molecules involved in signal transduction pathways and can shed light on mechanisms of metastasis and invasion of colon carcinoma cells. PMID- 9872567 TI - Saikosaponin b2 induces differentiation without growth inhibition in cultured B16 melanoma cells. AB - Treatment with 5 microM of saikosaponin (SS) b2 for 30 days was found to induce differentiation of B16 melanoma cells, with potentiation of expressions of melanogenesis and tyrosinase. To explore the mechanism of this effect, we observed the cell growth, cell cycle and morphology, and found that SSb2 did not affect any of these parameters. That is, SSb2 induced the differentiation of B16 melanoma cells without growth inhibition or cytotoxicity under conditions of low dose and long-term treatment. Phorbol ester, a protein kinase C (PKC) activator, markedly inhibited the expressions of melanogenesis and tyrosinase in both the control B16 melanoma cells and the long-term treated B16 melanoma cells. Down regulation of the PKC activity may be involved in the effects of SSb2. PMID- 9872568 TI - Recovery of flagellar inner-arm dynein and the fertilization tubule in Chlamydomonas ida5 mutant by transformation with actin genes. AB - The ida5 mutant of Chlamydomonas, first isolated as a mutant lacking a subset of axonemal inner-arm dyneins, has recently been shown to lack conventional actin owing to a serious mutation in its gene. It lacks inner-arm dyneins probably because actin is an essential subunit for their assembly. In addition, male gametes of ida5 are unable to produce the fertilization tubule, a structure that contains a core of actin filament bundles. To establish that those observed deficiencies are solely attributable to the loss of actin, and to provide a basis for future studies on the actin function in this organism, we examined in this study whether transformation of this mutant with cloned actin genes can rescue the mutant phenotypes. Cotransformation of the double mutant ida5arg2 with the wild-type actin gene and arginino-succinate lyase gene that suppresses the arg2 mutation yielded several transformants that displayed increased motility. All of them were found to have acquired the introduced actin gene in the genome and the product actin in the flagella, and regained the missing inner-arm dyneins and wild-type motility. In addition, most transformants also became able to grow the fertilization tubule when mating reaction was induced. In addition to the wild type actin gene, we also used a chimeric actin gene in which the N-terminal 12 amino-acid sequence of Chlamydomonas actin was replaced by that of the greatly divergent Tetrahymena actin. Transformants with this gene also resulted in recovery of inner-arm dynein and 70-80% of the wild-type level of motility. These results established that the lack of inner-arm dynein and the fertilization tubule in ida5 are consequences of its loss of conventional actin. Furthermore, they demonstrate that Chlamydomonas offers an excellent experimental system with which to study the structure-function relationship of actin by means of mutant analysis. PMID- 9872569 TI - Actin-filaments localize on the sorting endosomes of 3Y1 fibroblastic cells. AB - By immunofluorescence microscopic observation, monoclonal and polyclonal antibodies against a synthetic actin C-terminal peptide were found to stain too colloguial, ambiguous punctuate structures distributed throughout the cytoplasm of 3Y1 cells, independently of actin stress fibers. Antibody against rab5, a small GTP binding protein of the sorting endosome, and anti-actin antibody co stained these punctuate structures. On the other hand, transferrin receptor, a well characterized maker of the sorting and recycling endosomes, colocalized with actin on the vesicular structures at the cell peripheral region but not at the perinuclear area where the recycling endosome localized. These observations suggest that actin molecules localize on the sorting endosomes. Tropomyosin, F actin binding protein, also colocalized with actin on the sorting endosomes. From these results, we proposed that actin-filaments with tropomyosin constitute the membrane skeleton on the sorting endosome surface. This article is the first report to show that actin-filaments localize on the intact endosomes. PMID- 9872570 TI - Inhibition of nuclear envelope reconstitution in Xenopus interphase egg extract by hemin. AB - Addition of hemin to the nuclear reconstitution system of Xenopus interphase egg extract using sperm head chromatin resulted in abnormal pseudonuclei exhibiting flattened membrane patches randomly distributed both on the surface and inside the nuclei. The structures that resembled nuclear pores were observed on these flattened membrane patch structures. Although the nucleosome structure was formed as revealed by the micrococcal nuclease digestion, the B-type lamin uptake into the nuclei was inhibited by hemin. Using heminagarose affinity chromatography, we isolated several hemin-binding proteins from fully reconstituted pseudonuclei. Some of the hemin-binding proteins bound concanavalin A (Con A). Comparison of hemin-binding proteins with those isolated from both fractions of supernatant and pellet separated by high speed centrifugation of the egg extract showed that the hemin-binding proteins of pseudonuclei were supplied from both fractions. The uptake of nuclear hemin-binding proteins did not occur in the incompletely reconstituted nuclei resulting from addition of excess sperm chromatin to the system. These results suggest that the hemin-binding proteins participate in the late steps of nuclear reconstitution during formation of the nuclear envelope. PMID- 9872571 TI - Biological activities of a novel lectin derived from silkworm faeces: characteristic changes of mouse peritoneal macrophages by the lectin. AB - A novel lectin derived from silkworm faeces, named NUE, activates phagocytosis of mouse peritoneal macrophages. At this time, significant morphological changes of the cells take place. NUE-treated macrophages formed projection-like neurocytes within 12 hr of treatment, and appeared somewhat flat in shape with the activation of cell growth. Associated with the morphological changes, actin was organized in dot-like structures corresponding to cell-substratum contact sites in NUE-treated macrophages. Vinculin, a cytoskeletal protein involved in microfilament-membrane interaction, formed doughnut-like rings matching the actin dots, called "podosomes". Furthermore, observation by interference reflection microscopy showed that NUE-treated macrophages adhered more strongly to the substratum at local areas. According to these changes, more proteins associated with cell-substratum contact sites became to detergent-resistant. It was shown that NUE changed adhesive form of mouse peritoneal macrophages structurally and qualitatively. PMID- 9872572 TI - Internal inclusions in brown eggs: relationships with fearfulness and stress. AB - Tonic immobility reaction and heterophil: lymphocyte ratio (indices of fearfulness and stress, respectively) were studied in hens laying eggs with or without internal inclusions (blood and meat spots). Two brown egg Spanish breed of chickens were used. There were no significant differences between groups in terms of tonic immobility duration. Birds of the group with internal inclusions appeared to be less fearful when defined by the number of inductions to achieve tonic immobility (1.4+/-0.1 vs 1.1+/-0.1; P < 0.01). Tonic immobility duration was significantly shorter in the group of hens with internal inclusions, when only those laying the abnormal egg the day before measurement of tonic immobility were included in this group (264+/-33 vs 395+/-30 s; P < 0.01). There was a significant difference in the heterophil:lymphocyte ratio between the two groups of hens (P < 0.01), mean value being significantly higher in the group with internal inclusions (0.43+/-0.02) than in the control group (0.33+/-0.02). Hens that laid eggs with internal inclusions had significant heterophilia (P < 0.01). Differences were consistent across the breeds. The incidence of internal inclusions was significantly greater (P < 0.001) in eggs with pink shells (an abnormality associated with the effect of stress). The results suggest that the laying of eggs with internal inclusions involves changes in fearfulness and stressfulness of hens. PMID- 9872573 TI - Ribosomal RNA gene copy number and nucleolar-size polymorphisms within and among chicken lines selected for enhanced growth. AB - Ribosomal (r) DNA genotypes (rRNA gene copy number) and nucleolar phenotypes (nucleoli number and size) were studied in dam and sire commercial broiler pure lines from three primary breeder sources. Thirteen lines were studied to determine whether directionally selected broiler pure lines contain higher numbers of rRNA genes than a control line unselected for performance traits. Eight of the 13 lines exhibited rRNA gene copy averages between 261 and 331 copies, three lines had averages between 365 and 380, and two lines had average copy numbers equal to or greater than 450 rRNA genes. The overall source copy number average from one breeder company exhibited a value (402 rRNA genes) significantly different from the control value (300 rRNA genes). Nucleoli number and relative-size were examined in 9 of the 13 lines to establish ploidy and determine the population incidence of nucleolar size polymorphisms. All of the individuals examined for nucleolar phenotype expressed two nucleoli, indicating that gene copy number variation in those lines was generally unrelated to haploidy, aneuploidy, or polyploidy. A high frequency of individuals exhibited nucleolar size polymorphisms (line values of 57 to 87%). The results suggest that multiple nucleolus organizer region (NOR) types are segregating within and among broiler pure lines and that these NOR types contain variable numbers of rRNA genes that differ in nucleogenesis capacity. PMID- 9872574 TI - Measurement of genetic variation within and between Japanese quail lines using DNA fingerprinting. AB - The objective of the present experiment was to study genetic variation within and among well-defined Japanese quail lines by DNA fingerprinting. The Japanese quail lines included a randombred control line (R1) and lines developed from R1 by divergent selection over 30 generations for 4-wk BW (HW, LW) and total plasma phosphorus (TPP) (HP, LP), a measure of yolk precursor in the blood. In addition, two sublines (HW-HP, HW-LP) of HW, developed in the ninth generation, were included in the analysis. Males of the sublines were selected for increased 4-wk BW whereas females were selected for increased (HW-HP) or decreased (HW-LP) TPP. Sixteen individual DNA samples per line were digested with HaeIII restriction enzyme and hybridized with Jeffreys' 33.6 probe. The DNA fingerprints were analyzed with computer programs designed to measure band sharing (BS). Within lines, BS ranged from 0.384 to 0.525. The BS within the R1 line was less than that of all selected lines, except for the HP and LP lines, indicating that, in general, selection had increased genetic homogeneity within the selected lines. Between lines, BS was less than within lines and the R1 line had the highest average level of BS (0.278) with the other lines. The BS between lines for the selected lines ranged from 0.230 to 0.308 with an average of 0.265. In the comparison of the R1 line with the selected lines, it appeared that selection for increased TPP or decreased BW may have influenced BS levels. The relationships of the HW line with its sublines (HW-HP and HW-LP) were not accurately predicted by the DNA fingerprinting technique used. All lines were separated, as indicated by the genetic distance between lines. PMID- 9872575 TI - Persistence of Salmonella enteritidis from one day of age until maturity in experimentally infected layer chickens. AB - In each of two replicate trials, 1-d-old chicks were inoculated orally with a phage type 13 Salmonella enteritidis isolate (resistant to nalidixic acid). Although S. enteritidis was found in the livers, spleens, and ceca of all sampled chicks at 1 wk postinoculation, colonization generally persisted beyond 4 wk postinoculation only in the ceca. Nearly half of the remaining hens were still shedding S. enteritidis in their feces at 24 wk of age, but only 1 of 62 hens laid eggs that were internally contaminated with S. enteritidis during the initial 4 to 6 wk of egg production. Chickens exposed to S. enteritidis shortly after hatching can apparently remain infected until maturity, at which time they might produce contaminated eggs or spread the infection to other susceptible, previously unexposed hens. PMID- 9872576 TI - Poultry welfare in North America: opportunities and challenges. PMID- 9872577 TI - Behavior and behavioral needs. AB - An understanding of behavior is important in any consideration of poultry welfare. Behavior is a good indicator of states of suffering such as fear, frustration, and pain. It might also be possible to use social interactions as indicators of welfare. The possibility of using "luxury" behavior, such as play and exploratory behavior, as an indicator of positive emotional states, requires investigation. Important welfare consequences arise from the fact that some behavior may be so strongly motivated as to constitute a "need". A behavioral need will arise in the case of behavior, such as nesting, that is controlled largely by internal factors, because these factors will be present no matter what type of environment is provided. Behavior triggered largely by external stimuli, such as responses to predators, will not give rise to a need if the external factors can be removed from the environment. Dustbathing is an example of behavior controlled by complex interactions between internal and external factors; the extent to which this constitutes a need is still being debated. If a behavioral need arises, then it is important that the environment provided allows it to be performed without damage to the performer or other birds. It should also be remembered that birds may need to perform behavior, including appetitive as well as consummatory elements, although the functional consequences of these are no longer required for survival. Finally, the performance of certain behavior leads to an increase in health or physical condition that improves welfare later in life. PMID- 9872578 TI - Rapid growth problems: ascites and skeletal deformities in broilers. AB - Over the last 40 yr, genetic selection for rapid growth and improved feed efficiency has been very effective in meat-type poultry. Combined with changes in the feed that have increased both the nutritional and physical density to encourage a high nutrient intake, growth rate has more than doubled. The effect of genetic selection for high muscle to bone ratio and high calorie intake of a ration that supplies all nutritional requirements causes significant mortality from cardiovascular disease. In the chicken, sudden death syndrome (flip-over) and pulmonary hypertension syndrome resulting in ascites are the most important. Ruptured aorta, spontaneous turkey cardiomyopathy (round heart), and cardiomyopathy causing sudden death produce high mortality in turkeys. Rapid growth induced by high nutrient intake alone can cause severe lameness, bone defects, and deformity, as these problems are seen in animals that have not been selected for rapid growth: dogs, horses, pigs, ratites and wild birds kept in zoologic gardens. In meat-type poultry, growth-related disease can be reduced or eliminated by reducing feed intake without affecting final body weight. Rapid growth alone may not be the pathogenic mechanism that results in cardiovascular or musculoskeletal defects. Metabolic imbalance induced by high nutrient intake may cause some of the conditions. These metabolic problems might be corrected without reducing growth rate. PMID- 9872579 TI - Improving animal well-being through genetic selection. AB - This paper reviews the possibilities of adapting laying hens to cages by means of genetic selection. By selecting separately for rate of lay and longevity using a kin selection method, a strain of laying hen has been developed that shows much less feather pecking and cannibalism than a control strain, and with no decrease in productivity. This experimental strain enjoys a higher level of welfare in cages because it does not require beak trimming. PMID- 9872580 TI - Health and husbandry considerations of induced molting. AB - There have been many methods proposed to induce molting. Some worked very well in practice, but others were detrimental to the health and welfare of the hens. The most effective methods use some level of feed restriction and daylength manipulation to reduce body weight (Hansen, 1966; Ruszler, 1974, 1984, 1996; Swanson and Bell, 1974; Brake and Carey, 1983). Weight reduction is necessary for rest and rejuvenation of body tissues. Other methods evaluated incorporated dietary imbalances using either zinc, iodine, or sodium. Pharmaceuticals have been used but have not been cost effective. In recent years there have been those who question whether molting techniques are humane. Therefore, interest has been heightened in alternate methods to induce molting. Research reported to date has been inadequate to accurately determine which methods of induced molting are the least stressful, if they in fact, cause any more stress than that experienced by the hen during a natural molt. The three or four most highly refined methods being used commercially are not generally detrimental to the health and welfare of today's laying hen, provided that they are managed in accordance with proper husbandry practices. PMID- 9872581 TI - Mechanical harvesting of broilers. AB - At the end of the growout phase of broiler production essentially all broilers are caught and loaded into coops or cages entirely by hand. The catching of broilers remains one of the few aspects of broiler production not yet automated. Vacuum systems, permanent conveyors recessed in the floors of growout facilities, collecting mats, scoops, and self-propelled, combine-type devices have been tried with little success over the past 30 yr. Renewed efforts at developing mechanical broiler harvesters have been encouraging in recent years. Several equipment companies in North America and Europe have developed prototype mechanical harvesters that appear to have promise. The benefits of mechanical harvesting as compared to hand catching include lower costs and improved working conditions. In addition, studies conducted thus far indicate that mechanical harvesting will improve bird welfare both from a stress and injury standpoint. PMID- 9872582 TI - The problem of broken bones during the handling of laying hens--a review. AB - The major welfare concern during the handling of laying hens is that of broken bones. With particular reference to the United Kingdom, this paper reviews the work that has been done to quantify the problem, to examine the causes, and to investigate factors that affect it. The number of freshly broken bones found in live birds prior to slaughter and the number of old healed breaks found at slaughter are unacceptably high. End-of-lay hens from battery cages have especially fragile bones and these are easily broken during the rough handling that is received during depopulation. Birds from more extensive laying systems have stronger bones and suffer fewer breaks during depopulation but have a greater prevalence of old healed breaks. The old breaks occur as a result of collisions due to poor design within these housing systems. The number of fresh breaks can be reduced by increasing bone strength and handling birds with more care. The numbers of old breaks can be reduced by better design of housing systems and the physical environment within them. PMID- 9872583 TI - Physiological stress and welfare of broiler chickens in transit: solutions not problems! AB - The rearing of large numbers of broiler chickens at geographically dispersed sites means that they have to be transported by road, over various distances, to centralized processing plants for slaughter. The birds may be exposed to a variety of stressors during transit, including the thermal demands of the transport microenvironment. The thermal environments experienced by broiler chickens during routine transport in the U.K. on a large number of commercial vehicles under a wide range of external climatic conditions have been characterized using three-dimensional thermal mapping (temperature and water vapor density). Inadequate ventilation results in heterogeneous distributions of temperature and humidity and, thus, thermal loads within the vehicle, and, therefore, the existence of a "thermal core" in which the risk of heat stress is increased. Relationships between specific physiological indices of stress and quantified thermal loads have been determined in accurate transport simulations in the laboratory. The findings have been employed to establish a predictive model of the induction of heat stress during commercial transportation, as well as to define the acceptable ranges and limits for temperature and humidity within the transport containers. These principles have been utilized in developing a monitoring system to warn of impending heat stress and in improving vehicle design to facilitate the prevention of heat stress during broiler transportation. PMID- 9872584 TI - Welfare during stunning and slaughter of poultry. AB - This paper describes research on the electrical stunning of poultry and the problems of achieving an effective humane stun with water bath stunners. The welfare and meat quality advantages of using gas mixtures to stun and kill birds are then described. The evidence strongly suggests that chickens and turkeys can be killed very humanely using either 90% argon in air or a mixture of 30% carbon dioxide and 60% argon in air. PMID- 9872585 TI - Health and production in improved cage designs. AB - This paper describes the effects of various cage designs on health characteristics such as skeleton strength, plumage and foot condition, mortality, and some production traits such as egg quality and feed consumption. Three major steps in improving cage design and in developing new cage designs are described. Firstly, cage design in general has been improved by comparing different conventional commercial cage designs under the same experimental conditions. Secondly, unconventional design features, such as an abrasive strip to reduce excessive growth of claws or a perch to improve the behavioral repertoire for the hens as well as to strengthen skeletal structure, are described. Thirdly, the effects of fully furnished cages that also include a nest and dustbath for smaller or larger groups of birds are discussed. These designs create an environment in which the problems of conventional cages, such as behavioral restriction, are reduced and the shortcomings of large litter aviaries, such as cannibalism, parasites, and a poor working environment, are improved. Cages with one perch level containing groups of 4 to 10 hens seem more likely alternatives to conventional cages on larger scale farms than litter systems or colony cages for more than 10 birds. This is due mainly to the higher predictability of production, the decreased risk of cannibalism, and improved hygienic conditions. PMID- 9872586 TI - Modification of laying hen cages to improve behavior. AB - Although they have many disadvantages for welfare, including for behavior, laying hen cages also have advantages. For example, aggression and cannibalism (or the need for beak trimming to prevent these) are usually less than in other systems, benefiting both the birds and the producer. There have been three approaches to reducing other behavioral problems. First, the design of conventional cages has been improved, which has had favorable effects on some aspects of behavior, such as feeding. Second, cages for larger groups of birds have been tested; for example, the getaway cage has increased behavioral freedom but has also increased problems such as aggression and cannibalism. Third, novel cages have been designed for conventional group sizes. Many of the behavioral problems-for producers, birds, or both--occurring in conventional cages can be reduced or prevented by increasing cage area (including width) and height and by providing a nest box, dust bath, and perch. Some of these modifications could be implemented at negligible cost to the producer. Others will be commercially viable if premium prices are available for eggs or if legislation on housing of laying hens changes. PMID- 9872587 TI - Equipment design for breeding flocks. AB - Breeding stocks require specialized equipment and facilities that accommodate both sexes and their mating behaviors. Divergence in male and female requirements has occurred due to genetic selection, particularly in heavy meat stocks. Proper equipment design requires specific knowledge about the nutritional requirements, genetics, and behavior of the particular stock within the environment intended. Attention must be paid to the relationship between the rearing and breeding environments, because preferences and behavioral patterns develop at an early age. The "correct" breeding environment is highly dependent upon the growing environment. There is evidence that some undesirable behaviors and poor fertility may arise from inadequate rearing nutrition and that nutrition can influence the development of certain behaviors. PMID- 9872588 TI - B-complex associated immunity to Salmonella enteritidis challenge in congenic chickens. AB - Six trials were conducted during which a total of 12 congenic lines (University of California-Davis, UCD) homozygous for various B-complex haplotypes, were challenged as neonates by intraperitoneal injection with either of two isolates of Salmonella enteritidis. Because these B haplotypes were expressed on a common genetic background (highly inbred Line UCD 003), and mortality differences among lines were statistically significant in three of the six trials, and morbidity (body weight) differences were significant in another trial; it is suggested that B-complex alleles affect the degree of immunity to these isolates. When all lines and trials were compared, line 342 (BC/BC) emerged as particularly resistant, whereas lines 253 (B18/B18) and 254 (B15/B15) were more susceptible. The remainder of the lines were of neutral (intermediate) susceptibility. Sex did not appear to influence the results of the challenge, but more resistance was observed with an increase in the age at inoculation. Although the mechanism that determined this resistance is unknown it was present as early as 3 d of age, and it is suggested that complement proteins, which have a known role in protection from bacterial infections, and are encoded by genes located within the B-complex, or acute phase proteins, may account for these observations. The results provide additional evidence for the importance of the B-complex in determining immunity to Salmonella. PMID- 9872589 TI - The utilization of dipeptides containing L-arginine by chicken macrophages. AB - L-Arginine is the precursor of NO, a cytotoxic agent of macrophages. Studies were carried out to determine whether dipeptides containing arginine can be utilized by lipopolysaccharide (LPS)-activated avian macrophages for NO production. A chicken macrophage cell line, the HD11 cell, was used in all experiments. Peptidase activities were observed in fetal bovine serum (FBS) and macrophage serum free medium (Mac-SFM). Therefore, the utilization of dipeptides by macrophages was examined using Dulbecco's modified Eagle medium (D-MEM), a chemically defined medium, in short-term culture without FBS. Nitrite accumulation in the culture medium was used as the indicator of NO production. At concentrations of 0.15 mM in the culture media, L-leucinyl-L-arginine was 89% as effective as L-arginine in providing substrate for NO production. L-Argininyl-L leucine was 38% as effective as L-arginine. The effectiveness increased to 93 and 58%, respectively, when the concentrations of dipeptides and arginine were 1.0 mM. Both values were slightly higher in a second experiment (97 and 70%, respectively). L-Lysine (10 mM) inhibited nitrite formation from all three sources of L-arginine. In studies of initial rates of transport by HD11 cells in Hanks Balanced Salts solution (HBSS), both L-argininyl-L-leucine and L-leucinyl-L arginine inhibited arginine uptake. As lysine and arginine share a common transporter for cationic amino acids and are known to compete for transport, these studies suggest that the peptides were hydrolyzed extracellularly, yielding arginine that was transported into the cell where it served as a substrate for NO synthesis. PMID- 9872590 TI - Chicken CD4, CD8alphabeta, and CD8alphaalpha T cell co-receptor molecules. AB - New knowledge has recently been obtained about the evolutionary conservation of CD4, CD8alphaalpha, and CD8alphabeta T cell receptor (TCR) co-receptor molecules between chicken and mammals. This conservation extends from biochemical structure and tissue distribution to function. Panels of monoclonal antibodies and polyclonal antisera against different epitopes of chicken CD8 and CD4 molecules have proven their value in several recent studies. Chicken CD8 allotypes and homozygous strains carrying these allotypes have been established and these strains provide excellent models for further studies. The extensive polymorphism of CD8alpha in chickens has not been observed in any other species, suggesting that CD8alpha and CD8beta have evolved under different selective pressure in the chicken. A large peripheral blood CD4+CD8+ T cell population in chicken resembles that observed in some human individuals but the inheritance of peripheral blood CD4CD8alphaalpha T cells in the chicken is a unique observation, which suggests the presence of a single gene responsible for CD8alpha, but not CD8beta, specific expression. Despite these unique findings in chicken, the data on CD4, CD8alphaalpha, and CD8alphabeta molecules show that they have evolved before the divergence of mammalian and avian branches from their reptilian ancestors. PMID- 9872591 TI - Evaluation of the efficacy of intraperitoneal immunization in reducing Salmonella typhimurium infection in chickens. AB - Conventional methods of parenteral immunization with killed bacterin vaccines have met with limited success in protecting the avian intestinal mucosa from pathogens such as Salmonella typhimurium. For mucosal vaccines to be successful they must be evaluated for their ability to stimulate local secretory immunoglobulin (SIgA) at the mucosal surface, which acts as the first line of defense against invading pathogens. Previously we have demonstrated the ability of i.p. immunization with nonreplicating antigen in an appropriate adjuvant to induce a primary immune response, which, after an oral booster immunization, stimulates enhanced intestinal IgA responses in chickens. In the experiments reported here we have applied this immunization protocol to vaccinate against S. typhimurium in chickens, and examined the protection provided against subsequent S. typhimurium challenge by placing vaccinated birds on seeded litter with cohabitant infected birds. Immunized+challenged birds displayed delayed onset of S. typhimurium infection, both at the mucosal surface and within the reticuloendothelial system. Elevated anti-S. typhimurium IgG and IgA titers were detected in serum after vaccination, which markedly increased after challenge, to levels higher than in control+challenged chickens. Anti-S. typhimurium IgA in bile and intestinal scrapings supernatant was also higher in the immunized+challenged birds than in the control+challenged birds 15 d after challenge. This study illustrates the potential for i.p. vaccination to induce a mucosal immune response to S. typhimurium in chickens, which, in the challenge model employed here, provided partial protection against intestinal challenge with the same pathogen and was reflected in deferred onset of bacterial infection and shedding. PMID- 9872592 TI - Effect of chemical composition of sunflower seed meal on its true metabolizable energy and amino acid digestibility. AB - The effect of chemical composition of sunflower seed meal (SFSM) on TMEn and true amino acid digestibility (TAAD) was studied. In Experiment 1, the excretion pattern of three SFSM samples force-fed (30 g) to 10 adult cockerels (Hy-Line) each was followed for 84 h to determine the time interval for complete excretion of SFSM. Type of SFSM did not affect the excretion pattern of DM and energy (P=0.438, and P=0.189, respectively). Dry matter and energy excreted every 12 h decreased linear and quadratically (P < 0.001) with collection time. No differences were found from 48 h collection time on. So, an excreta collection period of 48 h was considered adequate for determining the TMEn of SFSM. In Experiment 2, 135 adult cockerels were force-fed to determine the TMEn of 11 samples of SFSM. Type of SFSM affected TMEn (P < 0.001), which ranged from 1,558 to 2,023 kcal/kg DM for SFSM of 31 to 42% CP, respectively. The TMEn was highly correlated (P < 0.001) to hemicellulose (r=-0.90), acid detergent lignin (r= 0.84), neutral detergent fiber (r=-0.82), and CP (r=0.77). Four prediction equations are proposed, the most practical being: TMEn (kcal/kg DM)=2,816.8 - 109.5 hemicellulose (%DM), RSD=70.2. Three out of the 11 samples of SFSM were selected for determining TAAD and the effect of endogenous amino acid correction. The methodology used was that of the TMEn assay, but one more estimation of amino acid endogenous excretion was made using a N-free diet with 85% cornstarch and 15% cellulose. Endogenous amino acid excretion was greater for roosters fed the N free diet than those deprived of feed, resulting in a higher digestibility (from 0.7 to 2.7%, P < 0.05) only for six amino acids: threonine, valine, alanine, proline, and aspartic and glutamic acids. No interaction was detected (P=0.94) between type of SFSM and method of estimation of endogenous amino acid excretion. The TAAD of SFSM increased significantly (P < 0.001) with the CP content, total TAAD being 86, 88, and 89% for SFSM of 32, 35, and 37% CP, respectively. Attention should be paid when including high fiber-low protein SFSM in poultry diets to balance its lesser digestible amino acids contribution, mainly in lysine (from 0.77 to 1.06% for SFSM of 32 and 37% CP, respectively). PMID- 9872593 TI - Lipopolysaccharide-induced reductions in body weight gain and feed intake do not reduce the efficiency of arginine utilization for whole-body protein accretion in the chick. AB - The effects of repeated injections of 400 microg Escherichia coli lipopolysaccharide (LPS) on chick performance from 11 to 22 d posthatching were examined in chicks fed casein-based diets containing graded levels of arginine. Administration of LPS reduced (P < 0.05) weight gain, feed intake, and protein accretion, and there was a tendency (P=0.07) for LPS administration to be more growth-depressing at the higher than at the lower levels of supplemental arginine. Regression analysis of protein accretion for the first three doses of arginine indicated that protein accretion was a linear (P < 0.01) function of supplemental arginine intake for both saline-injected (r2=0.94) and LPS-injected (r2=0.93) chicks. Slopes of the best-fit regression lines for both treatment groups were equal, indicating that arginine utilization for protein accretion was not affected by LPS administration. The dietary arginine concentration required to maximize weight gain and feed efficiency was unaffected by LPS administration, with both saline- and LPS-injected chicks reaching plateaus in weight gain and feed efficiency at 0.90 and 0.98% digestible arginine, respectively. PMID- 9872594 TI - Effect of dietary phytase and high available phosphorus corn on broiler chicken performance. AB - Two trials were conducted to determine the effects on broiler chicken performance and health of reducing dietary phosphorus levels by treating feed with the enzyme phytase, formulating diets using high available phosphorus (HAP) corn, or when diets were formulated with HAP corn and treated with phytase. Cobb x Cobb male broiler chickens were placed in an experimental design consisting of four dietary treatments with six replicate pens of 50 broilers per pen. The dietary treatments consisted of untreated control feed, phytase-supplemented feed (500 U/kg), diets prepared with HAP corn, and diets prepared with HAP corn and supplemented with phytase. The chickens were maintained on these dietary treatments from 1 to 49 d of age with feed and water made available for ad libitum consumption. When the two trials were combined, there was a significant (P < or = 0.05) increase in body weight in the broilers fed the phytase treated diets at 49 d of age. The serum activity of alkaline phosphatase was significantly decreased in the diets supplemented with phytase, and serum cholesterol was significantly decreased in the diets prepared with HAP corn. These data indicate that total phosphorus can be reduced by at least 11% in diets prepared with HAP corn, or in diets supplemented with phytase, without affecting the performance or health of broiler chickens. When diets are prepared with HAP corn and supplemented with phytase, the dietary addition of total phosphorus can be reduced by at least 25% without affecting broiler chicken performance or health. PMID- 9872595 TI - Lung lining fluid antioxidants in male broilers: age-related changes under thermoneutral and cold temperature conditions. AB - The purpose of this study was to determine age-related changes in lung lining fluid antioxidants in broilers reared under thermoneutral or cold temperature conditions. Male broilers (Cobb 500) were placed in floor pens within environmental chambers and fed a standard commercial starter diet. The thermoneutral Control chamber was maintained at 32, 30, 27, and 22 to 25 C for Weeks 1, 2, 3, and 4 to 7, respectively, whereas temperature in the Cold chamber was lowered to 18 C during Week 3 and maintained between 15 and 18 C for the rest of the study. At 2, 4, and 7 wk, four to six birds per chamber were selected randomly. The lungs were lavaged with heparinized saline (2 mL/g lung) to obtain lung lining fluid. Antioxidants [reduced (GSH), oxidized (GSSG), and total (TGSH) glutathione, uric acid, ascorbic acid, and alpha- and gamma-tocopherol] in lung lining fluid were determined by HPLC; protein was determined colorimetrically. In Controls, levels of alpha- and -gamma-tocopherol, uric acid, and GSH in lung lining fluid decreased between 2 and 7 wk of age. Birds in the Cold chamber exhibited higher protein, a higher GSSG:TGSH ratio, and a decrease in ascorbic acid (7 wk) in lung lining fluid relative to Controls. Lung lining fluid antioxidants were not correlated with antioxidants in plasma. To determine the effect of vitamin E supplementation on lung lining fluid antioxidants, birds were given a supplement of 200 IU alpha-tocopherol per day for 7 d. Alpha-tocopherol supplementation elevated alpha-tocopherol levels in lung lining fluid, but lowered ascorbic acid, GSH, and GSSG and had no effect on uric acid in lung lining fluid. The results of this study suggest that antioxidant protection in lung lining fluid may diminish with age, that cold conditions in this study produced an oxidative stress in lung lining fluid in broilers, and that oral supplementation of alpha-tocopherol elevated lung lining fluid alpha-tocopherol. PMID- 9872596 TI - In ovo administration of recombinant human insulin-like growth factor-I alters postnatal growth and development of the broiler chicken. AB - Two experiments assessed the efficacy of in ovo administration of insulin-like growth factor-I (IGF-I) to enhance skeletal muscle development and improve feed efficiency of broilers. Hatching eggs were divided into three groups: uninjected control, vehicle-injected control, and recombinant human (rh) IGF-I (100 ng per embryo). Eggs in Experiment 1 were injected on Day 1, 4, or one of Day 7 through 18 of incubation. Growth rates for Days 1 and 4 resulted in the greatest response to treatment (P < 0.01, P < 0.06 respectively). Based on these results, Experiment 2 focused on Days 1 to 4 of incubation. Results from Experiment 2 showed that there was no significant difference in hatchability among control and rh IGF-I treatment groups. Injection on Day 3 resulted in the greatest response for increased live (P < 0.035) and leg (P < 0.02) weights in both sexes. Feed efficiencies of all rh IGF-I groups were significantly (P < 0.01) improved for the first 3 wk. In ovo administration of rh IGF-I on Day 3 increased feed efficiency (6.65%; P < 0.009) in pens of mixed-sex broilers. In addition, live weights (12.3%; P < 0.002), leg weights (11.7%; P < 0.01), breast weights (9.9%; P < 0.04), and heart weights (11.4%; P < 0.02) were increased in males. These results demonstrate that in ovo administration of rh IGF-I alters feed efficiency, growth, and tissue development. This finding lends itself to significant improvements in broiler production efficiency and profitability. PMID- 9872597 TI - Regulation of matrix metalloproteinase-2 (gelatinase A, MMP-2), membrane-type matrix metalloproteinase-1 (MT1-MMP) and tissue inhibitor of metalloproteinases-2 (TIMP-2) expression by elastin-derived peptides in human HT-1080 fibrosarcoma cell line. AB - Soluble kappa-elastin peptides were shown to stimulate the expression of MMP-2 (but not MMP-9) by human fibrosarcoma HT-1080 cells, both at the protein and mRNA levels; maximal effect being observed at a concentration of 25 microg/ml of kappa elastin. The stimulatory effect could be reproduced using Val-Gly-Val-Ala-Pro-Gly (VGVAPG) peptide, an elastin-derived hydrophobic hexapeptide which represented the elastin receptor binding sequence of tropoelastin. Furthermore, treatment of cells with lactose (30 mM), which dissociated 67-kDa elastin binding protein (EBP) from cell surfaces, completely abolished this effect, suggesting that the elastin receptor could mediate such a response. Using a specific monoclonal antibody, 67-kDa EBP was detected in HT-1080 membrane preparations by Western immunoblotting. Following treatment with 25 microg/ml kappa-elastin or 200 microg/ml VGVAPG, increased levels of the active 62-kDa form of MMP-2 were found in HT-1080 cell extracts. Stimulation of MT1-MMP mRNA expression by treatment with elastin-derived peptides (EDPs) was shown by competitive polymerase chain reaction (PCR). A reverse zymography analysis revealed that EDPs also stimulated TIMP-2 (but not TIMP-1) production by HT-1080 cells. Competitive PCR confirmed increased TIMP-2 mRNA expression by such treatment. These results suggest that occupancy of the 67-kDa elastin receptor by elastin-derived peptides enhanced both expression and activation of proMMP-2 and consequently, could promote the invasive/metastatic ability of tumor cells expressing this receptor. PMID- 9872599 TI - Plasminogen activator system modulates invasive capacity and proliferation in prostatic tumor cells. AB - The malignant phenotype of prostatic tumor cells correlates with the expression of both uPA and its cell-membrane receptor (uPAR); however, there is little information concerning the role of cell-bound uPA in matrix degradation and invasion. Our results suggest that cell-associated uPA plays a key role in regulating the amount of plasmin present at the surface of prostatic carcinoma (PRCA) cells and show that differential production of uPA corresponds with the capacity to bind and activate plasminogen. In addition, we provide direct evidence that both uPA secretion and the presence of uPA-uPAR complexes characterize the invasive phenotype of PRCA cells and suggest the existence of several pathways by which tumor cells acquire plasmin activity. LNCaP cells (which do not produce uPA but express uPAR) may activate plasmin through exogenous uPA. In vivo, the source of uPA may be infiltrating macrophages and/or fibroblasts as observed in several other systems. PAI-1 accumulation in the conditioned medium (CM) limits plasmin action to the pericellular microenvironment. Our results indicate that MMP-9 and MMP-2 are also activated by plasmin generated by cell-bound but not by soluble, extracellular uPA. Plasmin activation and triggering of the proteolytic cascade involved in Matrigel invasion is blocked by antibodies against uPA (especially by anti- A-chain of uPA which interacts with uPAR) and by PA inhibitors such as p-aminobenzamidine which may regulate levels of cell-bound uPA. uPA may also regulate growth in PRCA cells. Indeed, antibodies against uPA A-chain (and also p-aminobenzamidine treatment) interfere with the ATF domain and inhibit cell growth in uPA-producing PC3 and DU145 prostate cancer cell lines, whereas exogenous uPA (HMW-uPA with ATF) induces growth of LNCaP prostate tumor cell line. These data support the hypothesis that in prostatic cancer patients at risk of progression, uPA/plasmin blockade may be of therapeutic value by blocking both growth of the primary tumor and dissemination of metastatic cells. PMID- 9872598 TI - Proteolysis of extracellular matrix by invadopodia facilitates human breast cancer cell invasion and is mediated by matrix metalloproteinases. AB - Breast cancer cell lines vary in invasive behavior and one highly invasive cell line (MDA-MB-231) proteolytically degrades extracellular matrix with invadopodia (Thompson et al. 1992, J Cell Physiol, 150, 534-44; Chen et al 1994, Breast Cancer Res Treat, 31, 217-26). Invadopodial proteolysis of extracellular matrix is thought to be necessary for invasion; however, this has not been demonstrated directly. To obtain such evidence, normal (HBL-100) and malignant (MCF-7, MDA-MB 231) breast cells were evaluated for invadopodial proteolysis of extracellular matrix and invasive behavior. We report that invadopodial proteolysis of immobilized fibronectin is positively correlated with invasion of cells into type I collagen gels. Moreover, reducing the proteolytic activity of invadopodia with the metalloproteinase inhibitor, batimastat (BB-94), also decreases invasion indicating that breast cancer cell invasion is dependent upon proteolytically active invadopodia. PMID- 9872600 TI - Platelet-derived growth factor (PDGF) BB acts as a chemoattractant for human malignant mesothelioma cells via PDGF receptor beta-integrin alpha3beta1 interaction. AB - Platelet-derived growth factor BB (PDGF BB) and the PDGF receptor beta are expressed on mesothelioma cells, but their biological function has not yet been defined. In the present study we used Boyden chambers fitted with filters coated with the adhesive matrix proteins fibronectin, laminin, collagen type IV or the nonmatrix adhesive molecule poly-L-lysine (PLL). Mesothelioma cells migrated towards PDGF BB at concentrations ranging from 0.78 to 12.5 ng/ml if matrix proteins were present as adhesive substrates. This migration was integrin dependent since the same cells failed to migrate if the adhesive interactions necessary for migration were provided by molecules other than integrins. Migration of mesothelioma cells on fibronectin, laminin or collagen-type IV in response to PDGF BB was inhibited if the cells were pretreated with blocking antibodies to alpha3beta1 integrin. These findings describe for the first time PDGF BB as a chemoattractant for malignant mesothelioma cells and that collaboration between PDGF receptor beta and integrin alpha3beta1 is necessary for the motile response of these cells to PDGF BB. PMID- 9872602 TI - G-CSF increases secretion of urokinase-type plasminogen activator by human lung cancer cells. AB - We reported previously that granulocyte colony-stimulating factor (G-CSF) can promote the invasion of human lung cancer cell lines in vitro. However, the exact mechanism of its stimulatory effect on invasion remains to be elucidated. In the present study we mainly focused our attention on the components of the plasminogen activation system in human lung cancer cell lines, because of the central role that plasminogen activators play in regulating extracellular proteolysis. We showed that G-CSF induced a dose-dependent increase in the urokinase-type plasminogen activator (uPA) activity in the conditioned medium of a PC-9 lung cancer cell line. When the amounts of uPA activity were quantitated by densitometry, we found that even at a concentration of 0.01 microg/ml, G-CSF had a stimulatory effect on the uPA release, while high concentrations caused a 3.6-fold increase at a maximum concentration of 1 microg/ml. A Western blot analysis of the conditioned medium confirmed the findings observed in a zymographic analysis. The observed increase in uPA protein was paralleled by a significant increase in the uPA mRNA levels after treatment with G-CSF. However, our experiments failed to identify any alteration in the plasminogen activator inhibitor (PAI) secretion caused by G-CSF. In addition, we also found the expression of G-CSF receptor by PC-9 cells, suggesting the possible pathway activated by G-CSF. PMID- 9872601 TI - Inhibition of tumor invasion and metastasis by calcium spirulan (Ca-SP), a novel sulfated polysaccharide derived from a blue-green alga, Spirulina platensis. AB - We have investigated the effect of calcium spirulan (Ca-SP) isolated from a blue green alga, Spirulina platensis, which is a sulfated polysaccharide chelating calcium and mainly composed of rhamnose, on invasion of B16-BL6 melanoma, Colon 26 M3.1 carcinoma and HT-1080 fibrosarcoma cells through reconstituted basement membrane (Matrigel). Ca-SP significantly inhibited the invasion of these tumor cells through Matrigel/fibronectin-coated filters. Ca-SP also inhibited the haptotactic migration of tumor cells to laminin, but it had no effect on that to fibronectin. Ca-SP prevented the adhesion of B16-BL6 cells to Matrigel and laminin substrates but did not affect the adhesion to fibronectin. The pretreatment of tumor cells with Ca-SP inhibited the adhesion to laminin, while the pretreatment of laminin substrates did not. Ca-SP had no effect on the production and activation of type IV collagenase in gelatin zymography. In contrast, Ca-SP significantly inhibited degradation of heparan sulfate by purified heparanase. The experimental lung metastasis was significantly reduced by co-injection of B16-BL6 cells with Ca-SP. Seven intermittent i.v. injections of 100 microg of Ca-SP caused a marked decrease of lung tumor colonization of B16 BL6 cells in a spontaneous lung metastasis model. These results suggest that Ca SP, a novel sulfated polysaccharide, could reduce the lung metastasis of B16-BL6 melanoma cells, by inhibiting the tumor invasion of basement membrane probably through the prevention of the adhesion and migration of tumor cells to laminin substrate and of the heparanase activity. PMID- 9872604 TI - Comparison of 16 human colon carcinoma cell lines for their expression of sialyl LeX antigens and their E-selectin-dependent adhesion. AB - Two anti-sialyl Lewis X (sLeX) monoclonal antibodies, mAb FH6 and mAb KM93, were analyzed by flow cytometry for their ability to bind to 16 human colon carcinoma cells. The binding profiles of these two anti-sLeX monoclonal antibodies did not correspond to each other. Three of the cell lines were reactive with mAb FH6 but not with mAb KM93. These three cell lines did not adhere to Chinese hamster ovary cells that were stably transfected with human E-selectin cDNA in an E-selectin dependent manner. In contrast, almost all human colon carcinoma cell lines that bound to mAb KM93 adhered to cells that expressed E-selectin. These results suggest that a subtype of sLeX carbohydrate epitopes recognized by mAb FH6 do not always function as ligands for E-selectin. PMID- 9872605 TI - Exercise training in heart failure. AB - Patients with heart failure challenge the clinician with a constellation of difficult clinical, pathophysiologic, and psychologic issues. As a result, until recently, exercise training was not considered a safe and effective treatment strategy to be used in these patients. However, in the past 10 years, data from both randomized and nonrandomized trials showed that regular exercise training in patients with stable Class II and III heart failure can safely improve exercise tolerance, attenuate an overactivated sympathetic nervous system, partially reverse skeletal muscle abnormalities, and enhance health-related quality of life. These outcomes are achievable with a relatively moderate dose of physical activity, such as 30 to 60 minutes of walking or cycling 3 to 5 days per week at an intensity equivalent to 60% to 70% of peak oxygen consumption. Sufficiently powered trials are needed to assess morbidity, mortality, and cost-effectiveness endpoints relative to exercise training in patients with heart failure. PMID- 9872606 TI - Calcium channel blockers in cardiac failure. AB - Congestive cardiac failure is an increasingly prevalent syndrome associated with a high morbidity and mortality. The role of calcium channel blockers in the treatment of heart failure is unclear. The potential benefits of these agents derive not only from their vasodilator properties, but also from anti-ischemic effects, beneficial effects on endothelial function and the development of atherosclerosis, and favorable effects on calcium cycling at a molecular level. Pitted against this array of potential benefits are direct negative inotropic effects and the potential for neuroendocrine activation. Treatment with short acting dihydropyridine agents has not resulted in long-term clinical benefits in patients with cardiac failure. Diltiazem may be beneficial in patients with nonischemic heart failure, and verapamil has a neutral effect in cardiac failure, although it may have a role in combination with ace inhibition. To date, amlodipine has been associated with the most promising results, with evidence of a mortality benefit in nonischemic heart failure. Mibefradil is of no benefit in the management of heart failure, although the trend toward increased mortality in the treatment arm of the Mortality Assessment in Congestive Heart Failure (MACH) 1 trial may have been due to drug interactions. The potential role of calcium blockers in diastolic dysfunction and in combination with ace-inhibition requires further study. PMID- 9872603 TI - Modulation of angiogenesis by human glioma xenograft models that differentially express vascular endothelial growth factor. AB - To evaluate the potential actions of vascular endothelial growth factor (VEGF) on capillary permeability and drug transport, tumorigenic human glioma cell lines were developed that expressed different levels of VEGF. Three human glioma cell lines (i.e. U373, SF126, SF188) were screened for VEGF under normoxic and hypoxic (i.e. induced by CoCl2) conditions by Western blot analysis. Subsequent to these results, sense and antisense VEGF164 cDNA transfections were conducted. It was found that parental SF188 (SF188/V-) and SF188/V+ (sense transfected) cells could serve as an appropriate in vivo model based on their divergent levels of VEGF expression. Media derived from SF188/V+ cells stimulated endothelial cell growth by 30-60%, and enhanced endothelial cell clonogenicity by 5-10-fold compared to SF188/V- or empty vector transfected cells. Nude rats implanted with either SF188/V- or SF188/V+ cells subcutaneously and intracerebrally formed tumors, with those derived from SF188/V+ cells growing at a faster rate. Immunohistochemistry analysis indicated that the expression of VEGF and number of capillaries (factor VIII assay) were approximately 3-fold greater in SF188/V+ tumors compared to SF188/V- tumors. Pharmacological assays, such as measurements of cytotoxicity and DNA adducts, in SF188/V- and SF188/V+ cells treated with carmustine or temozolomide were similar. Therefore, other than differences in VEGF expression and growth in vivo, SF188/V- and SF188/V+ cells possess a similar phenotype, and can serve as models to evaluate the influence of VEGF on drug transport. PMID- 9872608 TI - Antiplatelet and anticoagulant therapy in the prevention of thromboemboli in chronic heart failure. AB - Thromboembolism is an important complication in patients with heart failure. Several recent clinical trials have established the efficacy of anticoagulant therapy in patients with heart failure and atrial fibrillation. There is renewed interest in examining the role of antiplatelet and anticoagulation therapy in patients with heart failure in sinus rhythm. There is a need to identify patients at risk for thromboembolism in heart failure. However, there are data to suggest that occult thromboembolic events may contribute to disease progression, ischemic events, and sudden cardiac death. This review summarizes the incidence, potential mechanism, and therapeutic approaches for management of thromboembolism in heart failure. PMID- 9872607 TI - Parenteral inotropic support for advanced congestive heart failure. AB - Parenterally administered positive inotropic agents remain an important component of the therapeutics of cardiac dysfunction and failure. Dobutamine, a catechol, remains the prototype of this drug group, but recently has been joined by the phosphodiesterase III inhibitor, milrinone. Compared with dobutamine, milrinone has greater vasodilating-unloading properties. The catecholamine, dopamine, is often used as a parenteral positive inotrope; but at moderate to high dose, it evokes considerable systemic vasoconstriction. At lower doses, dopamine appears to augment renal function. Levosimendan and toborinone, new compounds with several mechanisms of action, are under active clinical investigation and review for approval. Parenteral positive inotropic therapy is indicated for short-term (hours to days) treatment of cardiovascular decompensation secondary to ventricular systolic dysfunction, low-output heart failure. More prolonged or continuous infusion of one of these agents may be necessary as a "pharmacologic bridge" to cardiac transplantation, another definitive intervention, or more advanced, intense medical therapy. An occasional patient will require a continuous infusion via indwelling venous catheter and portable pump, simply to be able to be discharged from the hospital setting and function in the home environment. Intermittent parenteral inotropic therapy for chronic heart failure has provoked considerable controversy and passion among cardiologists and heart failure specialists; an attempt is made to present this topic in an objective manner. PMID- 9872609 TI - Arrhythmogenic right ventricular dysplasia/cardiomyopathy: review for the clinician. AB - Although hypertrophic obstructive cardiomyopathy remains the most common cause of sudden cardiac death in young people, rarer causes, such as arrhythmogenic right ventricular dysplasia (ARVD), are now being increasingly recognized to lead to sudden cardiac death in the younger population. Recent advances in the understanding of the genetic inheritance, etiopathogenesis, diagnosis, and treatment options of ARVD have prompted a lot of research in this form of right ventricular cardiomyopathy. The purpose of this report is to review the etiopathogenesis, clinical manifestations, diagnosis and treatment modalities for ARVD, and recent advances in the understanding of this disease entity. PMID- 9872610 TI - Transarterial chemoembolization and percutaneous ethanol injection therapy in patients with hepatocellular carcinoma. AB - Both transcatheter arterial chemoembolization (TACE) and percutaneous ethanol injection therapy (PEI) have proven their efficacy in patients with unresectable hepatocellular carcinoma (HCC): TACE mainly in large lesions or disseminated disease and PEI in solitary lesions smaller than 3 cm. Although severe complications have been observed with both methods, their incidence is low. In 1991, the combination therapy of initial TACE followed by multiple sessions of PEI was introduced, allowing the repeated percutaneous treatment of lesions larger than 4 cm. This approach has been shown to be more effective than TACE alone in lesions up to 8 cm in diameter. An increased incidence of serious side effects compared to either method alone has not been reported so far. PMID- 9872611 TI - The diagnosis of gluten sensitivity and coeliac disease--the two are not mutually inclusive. AB - The traditional definition of coeliac disease is inadequate because it includes only patients with abnormal small intestinal morphology. Gluten sensitivity is a systemic disorder whose common factor is an immune response to gluten in the context of the susceptible 'coeliac' HLA haplotype and possibly environmental triggers. Gluten sensitivity embraces traditional coeliac disease as well as subjects with normal small bowel morphology including latent coeliac disease, dermatitis herpetiformis, and symptomatic gluten intolerance. The diagnosis of gluten sensitivity and coeliac disease are not mutually inclusive. Small intestinal biopsy and clinical criteria are essential in diagnosing classical coeliac disease. IgA endomysial antibody is valuable in identifying gluten sensitivity and has particular value as a screening test. Serology should include total IgA levels to exclude selective IgA deficiency, a potential cause of false negative IgA endomysial antibody. A combination of histology, serology and clinical criteria will identify most cases of coeliac disease and gluten sensitivity. PMID- 9872612 TI - Hepatic infarction following percutaneous ethanol injection therapy for hepatocellular carcinoma. AB - We report on two patients who developed hepatic infarction after undergoing percutaneous ethanol injection therapy (PEIT) for hepatocellular carcinoma (HCC). In both cases, liver function parameters deteriorated immediately after the ethanol injection, and enhanced computed tomography images showed a wedge-shaped avascular low-density area due to hepatic infarction. In one patient, PEIT was performed for a nodule treated with transcatheter arterial infusion (TAI) using a suspension of styrene maleic acid neocarzinostatin (SMANCS) 4 weeks before. In the other patient, TAI with SMANCS had been carried out 14 months previously for a different nodule in the same segment where the nodule treated with PEIT was located. When PEIT is used for patients with HCC who have previously undergone TAI, especially with SMANCS, PEIT may induce hepatic infarction. PMID- 9872613 TI - Diagnosis of gluten-sensitive enteropathy: is exclusive reliance on histology appropriate? AB - OBJECTIVE: Coeliac disease is a prevalent disorder but frequently remains undiagnosed because of varied modes of clinical presentation. In this study, methods for the detection of coeliac disease were evaluated in a clinical practice setting. METHODS: Small intestinal histology, IgA anti-endomysial and IgG anti-gliadin antibody tests were performed on 441 unselected, consecutive patients under investigation for small intestinal disease. Response to treatment and other clinical events were monitored over the ensuing years. RESULTS: Untreated coeliac disease was diagnosed in 97 patients and was excluded in 344. At clinical presentation, the endomysial antibody test was positive in 84 of the 97 untreated coeliac patients (sensitivity 87%) and negative in 340 of the 344 non-coeliac patients (specificity 99%). A typical histological lesion was found in 83 of the 97 coeliac patients (sensitivity 86%) but was absent in all 344 non coeliacs (specificity 100%). The sensitivity of the gliadin antibody test was 69% and the specificity was 71%. CONCLUSIONS: In unselected patients attending a gastroenterology clinic, small bowel histology and endomysial antibody serology show similar predictive value in the diagnosis of coeliac disease. These results emphasize that a combination of clinical, histological and serological criteria are required for effective diagnosis of this disorder. Exclusive reliance on histology or serology will result in failure to make a diagnosis in a significant proportion of patients. PMID- 9872614 TI - Autoimmune thyroid diseases and coeliac disease. AB - BACKGROUND: Coeliac disease may be associated with a wide variety of diseases of known or suspected immunological aetiology. OBJECTIVE: To screen for both (a) the prevalence of coeliac disease in adults with autoimmune thyroid diseases, and (b) thyroid impairment among adults with coeliac disease, as compared to sex- and age matched controls. DESIGN: Prospective cohort study. SETTING: University teaching hospital. PATIENTS: A total of 152 consecutive adults with autoimmune thyroid diseases, 185 consecutive coeliac disease patients (53 newly diagnosed and 132 already on a gluten-free diet) and 170 sex- and age-matched controls. METHODS: Screening for coeliac disease was done by means of IgA anti-endomysium antibodies, detected by indirect immunofluorescence on monkey oesophagus. Patients with positive sera underwent duodenal biopsy for diagnostic confirmation. Thyroid function was assessed by measuring the levels of serum thyroid-stimulating hormone, free T3, free T4, thyroperoxidase and thyroid microsome antibodies. Autoimmune thyroid diseases were classified according to the American Thyroid Association guidelines. RESULTS: Anti-endomysium antibodies were positive in five of 152 autoimmune thyroid disease patients (3.3%) and coeliac disease was histologically confirmed in all: this prevalence is 10-fold higher than expected. Only one patient presented with gastrointestinal complaints, but iron deficiency was found in three and alterations at bone mineralometry in all. The overall prevalence of autoimmune thyroid diseases was significantly higher (38/185, 20.5%) in coeliac patients than in controls (19/170, 11.2%). The prevalence of both hypo- and hyperthyroidism was not different from that of controls, while the prevalence of autoimmune thyroid disease with euthyroidism was 13% in patients and 4.7% in controls. CONCLUSIONS: The association of coeliac disease with autoimmune thyroid disease is not surprising as they share common immunopathogenetic mechanisms. It is advisable to screen autoimmune thyroid disease patients for coeliac disease as there is an increased risk for gluten intolerance. In contrast, thyroid function assessment in coeliac disease patients is probably less justified, although the need for a strict clinical follow-up of those patients with euthyroidism and autoimmune thyroid disease, who could develop overt thyroid impairment, remains an open question. PMID- 9872616 TI - Oesophageal motility patterns and arterial blood pressure in patients with chest pain and normal coronary angiogram. AB - BACKGROUND AND OBJECTIVES: In previous manometric investigations, we observed that patients with chest pain and arterial hypertension frequently tend to display oesophageal motility abnormalities. Therefore, we set out to study this systematically. METHODS: Patients with chest pain and normal coronary angiogram (n=40) and healthy controls (n=20) were studied prospectively in a standardized fashion using a portable oesophageal manometry and blood pressure registration system over 24 hours. RESULTS: Twenty patients exhibited increased arterial blood pressure (24 h median > 135/85 mmHg), while in the other 20 patients and all controls the 24 h blood pressure patterns were normal. Median pressure amplitudes in the distal oesophagus were 46.5, 33 and 27 mmHg in patients with or without arterial hypertension and controls, respectively, and 30, 27 and 27 mmHg in the proximal oesophagus, respectively. The durations of distal contractions were 3.9, 3.4 and 3.4 s, respectively, and those of proximal contractions were 3.2, 3.0 and 3.2 s, respectively. Percentages of propulsive contractions were 53%, 44% and 59%, respectively, and those of simultaneous contractions were 23%, 25% and 10%, respectively. CONCLUSIONS: Patients and controls differ significantly regarding their oesophageal motility patterns. Patients without arterial hypertension exhibit impaired propulsion of oesophageal contractions, whereas patients with arterial hypertension tend to produce oesophageal hypermotility. This suggests that, depending on the presence or absence of arterial hypertension, different pathomechanisms of oesophageal motility disturbances come into play. PMID- 9872615 TI - A prospective evaluation of dermatological side-effects during alpha-interferon therapy for chronic viral hepatitis. AB - OBJECTIVE: Alpha-interferon therapy may occasionally account for immune-mediated phenomena. This study was conducted in an attempt to investigate the incidence of the development of immune-mediated dermatological diseases during alpha interferon therapy in patients with chronic viral hepatitis. The latter has not been evaluated prospectively, whereas most of the previous studies examined small numbers of interferon treated patients or consisted of case reports. DESIGN: A prospective case-control study. SETTING: A tertiary referral centre. PARTICIPANTS: One hundred and twenty consecutive patients with chronic viral hepatitis (67 with hepatitis B, 45 with hepatitis C, six with both hepatitis viruses, and two with delta hepatitis) were evaluated during a course of alpha interferon therapy. In addition, 120 consecutive patients with chronic liver diseases (disease control group), who had never received alpha-interferon therapy, were evaluated during the period of the study (at least for 12 months). INTERVENTIONS: Recombinant alpha-interferon at a dose of 4.5 or 5 million units subcutaneously (s.c.) three times per week for 6 to 12 months was administered to patients with hepatitis B. The patients with chronic hepatitis C were treated with 3 million units s.c. three times per week for 12 to 18 months. The patients with chronic hepatitis B and C infections received 4.5 million units for 6 months, and then 3 million units for an additional 6 to 12 months. Finally, the patients with chronic delta hepatitis received 5 million units for 1 year or more. MAIN OUTCOME MEASURES: To assess prospectively the incidence of these dermatological disorders during alpha-interferon therapy and to estimate if there is any relationship between their development and the clinical, laboratory or other characteristics of the patients with chronic hepatitis. RESULTS: Three to 6 months after the initiation of alpha-interferon three patients with chronic viral hepatitis (two with hepatitis C and one with hepatitis B) developed lichen planus, whereas one patient with hepatitis C developed relapsing aphthous stomatitis. The development of these disorders was significantly associated only with the presence of antinuclear antibodies before the initiation of alpha interferon (P=0.000000). None of the patients from the disease control group had such a manifestation during the follow-up. Lichen planus resolved after the end of therapy in all of them. In contrast, therapy was discontinued in the patient who developed aphthous stomatitis, owing to the painful lesions. CONCLUSIONS: This study demonstrated that alpha-interferon may rarely (3.3%) induce immune mediated dermatological disorders, especially lichen planus. The development of these disorders may reflect a subclinical or covert autoimmune background of patients, as suggested by the presence, although in low titres, of antinuclear antibodies. However, when lichen planus developed, it was mild, did not require the discontinuation of therapy and resolved after alpha-interferon administration had ceased. PMID- 9872617 TI - Gastrinomas and the change in their presentation and management in Northern Ireland, UK, from 1970 to 1996. AB - Thirty-five new cases of gastrinomas were diagnosed in N. Ireland between 1970 and 1996. Over this period, patient care has improved, with advances in imaging techniques and therapeutic regimens. Patients are now no longer presenting in the classical way with severe ulcer diathesis. Diarrhoea is often a major feature, occurring in 46% of patients. Thirty-one percent of patients presented with mixed amine precursor, uptake and decarboxylation (APUD) tumours. Survival has improved, most likely as a result of better detection of tumours, as well as treatment that is aimed at resection and removal of the gastrinoma. The advent of proton pump inhibitors has ensured symptom control in those for whom total tumour removal is impossible. Owing to improved survival, metastatic complications are often associated with patient mortality. PMID- 9872618 TI - A prospective study of the management of the young Helicobacter pylori negative dyspeptic patient--can gastroscopies be saved in clinical practice? AB - BACKGROUND: Helicobacter pylori status has been suggested as a means of selecting young dyspeptic patients for gastroscopy as patients who are H. pylori negative and do not exhibit alarm symptoms or ingest non-steroidal anti-inflammatory medication have a low risk of serious organic disease. AIM: To determine if young patients with ulcer-like dyspepsia and found to be H. pylori negative on non invasive testing could be reassured by this knowledge and not proceed to gastroscopy. PATIENTS: One hundred and sixty-one consecutive attendees aged 45 years or less with a presenting complaint of epigastric pain or discomfort were prospectively recruited from open access gastroscopy referrals and gastroenterology clinics. METHODS: Patients who were H. pylori negative on 13 carbon urea breath test were reassured of the likelihood of a normal gastroscopy, given lifestyle advice and also advised to take symptomatic therapy as required. Patients were reviewed at 6 weeks, 3 months and 6 months when symptoms and quality of life were reassessed. Patients proceeded to gastroscopy if at any review their dyspepsia score stayed the same or worsened. RESULTS: Fifty-five H. pylori negative patients were recruited (30 male, mean age 31 years), two patients did not attend subsequent review. Thirty-two (58%) came to gastroscopy. Endoscopic diagnoses included 25 which were normal, three with gastro-oesophageal reflux disease, three with peptic ulcer disease and one with gastric erosions. Dyspepsia and quality of life scores showed significant improvement over 6 months. CONCLUSIONS: This management strategy resulted in a 42% reduction in gastroscopies in H. pylori negative patients. Whilst the majority of patients endoscoped had normal findings, seven patients (22%) had pathology. Overall there were significant improvements in dyspepsia and quality of life at 6 month follow up. PMID- 9872619 TI - Results of Helicobacter pylori screening and eradication in a multi-ethnic community in central England. AB - OBJECTIVES: To determine the prevalence of Helicobacter pylori and risks for infection in a multi-ethnic inner city population, and to assess the effectiveness with which the infection can be eradicated in a relatively asymptomatic population of this type. DESIGN: Analysis of results from a small research-based community programme of screening and eradication. Tests were offered to matched groups of Asians and non-Asians from the general population. METHODS: Those who attended for screening had a serological screening test for infection with H. pylori. Data concerning possible risk factors for infection were collected by means of questionnaires. Subjects testing positive were offered a prescription for eradication therapy (ranitidine bismuth citrate and clarithromycin) through their general practitioner, and success of eradication therapy was assessed by urea breath test. RESULTS: The overall prevalence of infection in 155 subjects was found to be 51%. The difference between positive test rates in Asians (53%) and non-Asians (47%) was not significant (P=0.611). In Asians, age and social class were risk factors for infection, but not gender or birth outside the UK. Results were similar in those born in India and East Africa. Of the 79 people who tested positive, 64 (81%) elected to take eradication therapy. Compliance with medication was estimated to be 95% and successful eradication was achieved in 92% of cases. CONCLUSION: Social class is more important than Asian ethnic origin as a predictor of H. pylori status. Good rates of H. pylori eradication can be achieved in subjects from the general population living in inner city communities. PMID- 9872620 TI - Life threatening diarrhoea ultimately cured by surgery. AB - A 30-year-old previously healthy woman was diagnosed as having a vasoactive intestinal polypeptide (VIP)-producing tumour of the pancreas. Her medical history was typical for neuroendocrine gastrointestinal tumours, presenting initially with non-specific symptoms but eventually she developed life threatening manifestations requiring intensive care due to severe dehydration. She immediately recovered following surgical resection. The patient had elevated serum concentrations of VIP as well as pancreastatin, and post-operatively elevated concentrations of three growth factors, IGF-I, EGF and TGF-alpha, were seen. The importance of the alterations in plasma concentrations of the different peptides for her symptomatology are discussed. PMID- 9872621 TI - Endoscopic treatment in a case with extensive spontaneous intramural dissection of the oesophagus. AB - Spontaneous intramural dissection of the oesophagus has been recognized as a separate clinical entity between a transmural oesophageal rupture (Boerhaave syndrome) and an oesophageal mucosal tear (Mallory-Weiss syndrome). It is important to differentiate this condition from myocardiac infarction, dissecting aneurysm and other acute surgical conditions. Conservative management is usually thought to be adequate. We report a case of spontaneous intramural oesophageal dissection, in which the symptom of dysphagia did not improve with the conservative management and an endoscopic incision of the septum between true and false lumens using a needle-type diathermy knife was done safely and effectively. PMID- 9872622 TI - Pancreatic tuberculosis in non-immunocompromised patients: reports of two cases, and a literature review. AB - We present two cases of biopsy proven tuberculosis of the pancreas in non immunocompromised patients diagnosed and treated in our unit within the last 14 years. The first case presented with abdominal pain and fever, and the second with iron deficiency anaemia and severe weight loss. In both cases abdominal ultrasound and computed tomography suggested a pancreatic carcinoma. There was no pulmonary or intestinal tuberculosis. The tuberculin skin test was positive. Upon exploratory laparotomy the macroscopic appearance of the pancreas was that of an inoperable pancreatic carcinoma. Following the histological diagnosis of pancreatic tuberculosis, both patients were successfully treated with triple antituberculous therapy for 6 months. Isolated pancreatic tuberculosis is an extremely rare disease with only 41 cases in non-immunocompromised patients reported worldwide (1966-1997). It is a curable disease and should be considered in the differential diagnosis of a pancreatic mass or abscess shown on ultrasound or computed tomography, especially in developing countries, where tuberculosis is common. PMID- 9872623 TI - Pneumocystis carinii: a historical perspective. AB - The emergence of Pneumocystis carinii pneumonia is traced from its origins as an obscure pulmonary pathogen to its place as a prominent cause of lung infection in patients who have acquired immune deficiency syndrome. Improvements in diagnosis and treatment have resulted in better care of patients with this infection. In addition, basic immunologic and molecular investigations have improved our understanding of the nature of this organism and continue to provide important questions for further research. This is a US government work. There are no restrictions on its use. PMID- 9872624 TI - Current issues in the epidemiology, transmission, and reactivation of Pneumocystis carinii. AB - Issues related to the epidemiology, transmission, and reactivation of Pneumocystis carinii remain controversial. In this article, current studies related to geographic variations in the prevalence of P. carinii pneumonia are reviewed. In addition, extensive investigations show that exposure to P. carinii occurs early in life, followed by seemingly complete clearance of the organisms. Recent molecular techniques show that P. carinii pneumonia during periods of immune suppression can occur either by reactivation or by acquisition of new organisms. These studies are analyzed and a composite model of P. carinii infection is proposed. PMID- 9872625 TI - Pneumocystis pneumonia: clinical presentation and diagnosis in patients with and without acquired immune deficiency syndrome. AB - Pneumocystis carinii causes severe pneumonia in immunocompromised hosts. Although this most commonly occurs in patients with the acquired immunodeficiency syndrome (AIDS), other groups of immunocompromised patients without AIDS are also at risk for P. carinii pneumonia. These patients have solid or hematologic malignancies, organ transplantation, or inflammatory conditions requiring chronic immunosuppressive drugs, particularly corticosteroids. There are important differences in the clinical presentation of P. carinii pneumonia in patients with and without AIDS. P. carinii causes an acute fulminate pneumonia in patients without AIDS while patients with AIDS have more insidious involvement. The organism burden and lung inflammatory response are markedly different between these groups, contributing to substantial differences in clinical presentation, outcome, and mortality. The diagnosis of P. carinii pneumonia remains challenging for primary care physicians and specialists alike. The specific diagnosis of P. carinii pneumonia requires demonstration of the organism from a clinically relevant source, such as sputum, bronchoalveolar lavage fluid, or lung tissue. Alternative methods to diagnose P. carinii pneumonia are currently investigational and are an active area of research. The rapid and specific diagnosis of P. carinii pneumonia allows institution of specific treatment and improvement in patient outcome. PMID- 9872626 TI - Treatment and prophylaxis of Pneumocystis carinii pneumonia. AB - Early effective management of Pneumocystis carinii pneumonia improves outcome in patients with this disorder. Trimethoprim-sulfamethoxazole remains the agent of choice for treatment of severe P. carinii pneumonia. Pentamidine, trimethoprim dapsone, atovaquone, and other regimens are useful in selected clinical situations. Adjunctive corticosteroids are indicated in patients with acquired immune deficiency syndrome and P. carinii pneumonia who have moderate to severe P. carinii pneumonia defined as a room air arterial PaO2 less than 70 mm Hg or an alveolar-arterial oxygen gradient of greater than 35 mm Hg. The use of trimethoprim-sulfamethoxazole, dapsone, and aerosolized pentamidine in immunocompromised patients without AIDS is also reviewed. PMID- 9872627 TI - Taxonomy, genetic organization, and life cycle of Pneumocystis carinii. AB - Pneumocystis carinii was initially misidentified as a protozoan parasite. Recent molecular and biochemical analyses provide unequivocal evidence for placement of P. carinii with the fungi, and that P. carinii is most likely an ascomycete. Genetic investigations further show that P. carinii derived from different mammalian hosts (human, rat, mouse, and ferret) exhibit considerable chromosomal and gene sequence divergence indicating that they are likely of different species. The life cycle of P. carinii has not been definitively established, but available evidence is reviewed in light of classification of this organism as a fungus. PMID- 9872628 TI - Antigenic characterization of Pneumocystis carinii. AB - Studies of Pneumocystis carinii have been limited by our inability to propagate it in continuous culture. In this context, studies of P. carinii antigens have provided significant insight into the biology of this organism. The mannose-rich surface major surface glycoprotein of P. carinii termed glycoprotein A (gpA) is the best studied of these P. carinii antigens. Significant genetic and immunologic diversity exists between the gpA molecules expressed by P. carinii derived from different mammalian sources. The molecular and biochemical nature of gpA and other P. carinii antigens including p55 are reviewed. In addition, available information concerning the role of P. carinii gpA and other antigens in host-organism interactions are also discussed. PMID- 9872629 TI - Pneumocystis carinii interactions with respiratory epithelium. AB - Ultrastructural studies indicate that the attachment of Pneumocystis carinii to alveolar epithelial cells is a central feature during development of this pneumonia. P. carinii attaches preferentially to type I alveolar epithelial cells, and rarely binds to type II cells. Extensive in vitro investigations support roles for both fibronectin and vitronectin, which bind to the organism, and mediate P. carinii attachment to respiratory epithelial cells through corresponding integrin receptors. Additional studies indicate that P. carinii cytoskeletal rearrangement occurs after initial binding of the organism to lung cells. Further, the attachment of P. carinii to host epithelial cells is believed to induce ultrastructural changes in these cells and to result in enhanced epithelial cell permeability. Further studies will be required to determine the mechanisms and consequences of P. carinii attachment to host epithelial cells in the intact host. PMID- 9872630 TI - Lymphocytes in host defense against Pneumocystis carinii. AB - Host defense against Pneumocystis carinii depends on complex interactions between host immune cells and mediators. In immunocompetent hosts, the immune system provides efficient and effective defense against P. carinii. Clinical and experimental investigations confirm that lymphocytes control and coordinate this defense. During states of immunosuppression, however, lymphocyte function is impaired and clinical P. carinii pneumonia results. Lymphocytes participate in host defense by regulating other immune cells (CD4+ and CD8+ T cells), by producing antibodies against pathogens (B cells), and by killing of organisms (cytotoxic CD8+ T cells and natural killer cells). Animal models of P. carinii pneumonia, using animals with genetic or induced immunodeficiencies, have provided recent and relevant information about the roles of lymphocytes in host defense. The CD4+ T cell plays a central role in defense, and CD4+ T cells are impaired both quantitatively and qualitatively in immunosuppressed hosts. However, the mechanisms by which CD4+ T cells control defense against P. carinii require further investigation. When CD4+ T cells are unavailable for defense, CD8+ T cells can participate in defense against P. carinii, but the mechanisms of protection also remain to be determined. Although serum antibodies directed against P. carinii are ubiquitous in humans, including human immunodeficiency virus-infected individuals, recent experimental evidence shows that B cells and antibodies can contribute significantly to host defense against P. carinii. These data suggest that modulation of B cell function remains a valid approach to vaccine development. Overall, an improved understanding of lymphocytic defense against P. carinii could lead to rational development of immunotherapies directed against this important opportunistic pathogen. PMID- 9872632 TI - Pneumocystis carinii: the art and science of survival in the hostile environment of the alveolar spaces. AB - Pneumocystis carinii remains an important cause of pneumonia in immunocompromised patients, particularly in those who have acquired immune deficiency syndrome. Potential mechanisms by which P. carinii escapes elimination by lung defenses and proliferates are discussed. Evidence indicates that P. carinii attachment to alveolar epithelium is an important event in the establishment of infection and occurs through a variety of means. The binding of the organisms appears to be injurious to alveolar epithelial cells. Proliferation of the organism is associated with an ineffective and at times detrimental host inflammatory response. Additional studies indicate that P. carinii has the ability to invade tissues. Future investigations using new methodologies and strategies will be required to fully control this pathogen. PMID- 9872631 TI - Alveolar macrophage and glycoprotein responses to Pneumocystis carinii. AB - Pneumocystis carinii continues to represent an important complication of immunosuppression in patients with acquired immune deficiency syndrome, hematological and solid malignancies, organ transplantation, and during corticosteroid and cytotoxic therapy for inflammatory disorders. Although host defenses against this organism center around CD4 lymphocytic function, additional immune mediators in the alveolar spaces contribute substantially to host recognition and elimination of P. carinii. In particular, this review considers the interactions of P. carinii with alveolar macrophages, adhesive glycoproteins including vitronectin and fibronectin, and surfactant lipids and protein components. Recent studies indicate that alveolar macrophages contribute significantly to host responses against this organism by mediating uptake and degradation of P. carinii, and by releasing inflammatory mediators including reactive oxidants, eicosanoids, and the potent proinflammatory cytokine tumor necrosis factor-alpha. Furthermore, the interactions of P. carinii with multiple adhesive proteins and with surfactant components additionally modulate the interactions of P. carinii with macrophages and enhance host recognition of this pathogen. These non-lymphocytic mediators represent additional important mechanisms of host recognition and response to P. carinii infection. PMID- 9872633 TI - Diagnostic case study: varicella pneumonia. PMID- 9872634 TI - Long-term cardiac arrest survivors of the Vienna emergency medical service. AB - The purpose of this study was to describe the life of survivors after successful resuscitation and to see if there was an association with the type of emergency cardiac care. The 'Utstein-style' data of patients surviving non-traumatic cardiac arrest 24 (14-32) months were prospectively collected. The everyday activities and psychological concerns of patients with a cerebral performance category (CPC) of 1 and 2 using a questionnaire were analyzed. The chi2-square test was used for statistical analysis. The questionnaires of 92 patients (median age 59, IQR 51-68; females 36) were evaluated. Patients enjoy life (84%; n=73), have depression (36%; n=31), consider their survival a 'second chance' (84%; n=73) and fear that they may suffer cardiac arrest again (56%; n = 45). The average quality of life is 7 on a scale from 0 (worst) to 10 (perfect). The majority of cardiac arrest survivors have a satisfactory life. No significant correlation between the type of emergency cardiac care and post cardiac arrest life was found. The fact that there was no association with the type of emergency cardiac care may be due to the narrow selection of patients (CPC 1 and 2), the small number of patients or factors contributing to post cardiac arrest life other than emergency treatment. PMID- 9872635 TI - A revised role for the hospital cardiac arrest team? AB - This prospective study assesses the role of the hospital cardiac arrest team. We have determined who is responsible for return of spontaneous circulation in those patients who survive to discharge. A total of 83 adult patients were analysed over a 5 month period. Fifty-five patients had cardiopulmonary resuscitation after an in hospital cardiac arrest and circulation was restored in 14 (25.5%). Eight (14.5%) patients survived to discharge. All eight survivors had circulation restored by the first responder before arrival of the cardiac arrest team. Six survivors had witnessed ventricular fibrillation and successful defibrillation by nursing staff. The two other survivors had cardiac arrests in the presence of doctors. Twenty-eight patients had pre-hospital cardiac arrests and were attended by the cardiac arrest team in the emergency department. Six patients had circulation restored by the emergency medical services prior to hospital arrival and four (14%) survived to discharge. Two patients had circulation restored in the emergency department by the arrest team and none survived to discharge. All the survivors to hospital discharge had first responder return of spontaneous circulation. We suggest a revised role for the cardiac arrest team. PMID- 9872636 TI - Comparison of two paediatric resuscitation charts for ease of use. AB - We have compared the ease and speed of use of two published paediatric resuscitation charts, the Burke and Bowden chart and the Oakley chart. Twenty general anaesthetists each answered 11 questions on resuscitation data for five paediatric patients of different weights, heights and ages. Data was recorded for the five patients using one chart and then repeated for the same patients using the second with the order of use of chart being randomised (n=100 patients for each chart). There were significantly fewer patients for whom mistakes were made with the Burke and Bowden chart compared to the modified Oakley chart (21 and 46, respectively, P < 0.001). Only one physician recorded the resuscitation data correctly for all five patients with each chart. The Burke and Bowden chart was also statistically significantly faster to use than the Oakley chart (52.7 s (range 31-103) versus 64 s (range 44-115), respectively (P < 0.001)). A change in format and content of each chart may be required to make them both more user friendly especially when used under stress. PMID- 9872637 TI - Out-of-hospital cardiac arrests in Amsterdam and its surrounding areas: results from the Amsterdam resuscitation study (ARREST) in 'Utstein' style. AB - The purpose of this study was to describe the chain of survival in Amsterdam and its surroundings and to suggest areas for improvement. To ensure accurate data, collection was made by research personnel during the resuscitation, according to the Utstein recommendations. Between June 1, 1995 and August 1, 1997 all consecutive cardiac arrests were registered. Patient characteristics, resuscitation characteristics and time intervals were analyzed in relation to survival. From the 1046 arrests with a cardiac etiology and where resuscitation was attempted, 918 cases were not witnessed by EMS personnel. The analysis focussed on these 918 patients of whom 686 (75%) died during resuscitation, 148 (16%) died during hospital admission and 84 patients (9%) survived to hospital discharge. Patient and resuscitation characteristics associated with survival were: age, VF as initial rhythm, witnessed arrest and bystander CPR. EMS arrival time was significantly shorter for survivors (median 9 min) compared to non survivors (median 11 min). In 151 cases the police was also alerted and arrived 5 min (median) earlier than EMS personnel. Using the OPC/CPC good functional health was observed in 50% of the survivors and moderate performance in 29%. All links in the chain of survival must be strengthened, but equipping the police with semi automatic defibrillators may be the most useful intervention to improve survival. PMID- 9872638 TI - Treatment of out-of-hospital cardiac arrest in Ljubljana: outcome report according to the 'Utstein' style. AB - We investigated survival of patients with out-of-hospital cardiac arrest in Ljubljana according to the 'Utstein' style. Ljubljana consists of urban, suburban and semi-rural communities which encompass an area of 1615 km2 with 397306 residents. The area is served by a single response emergency medical system and local family practitioners. Between January 1, 1995 and December 31, 1997 cardiac arrest was confirmed in 966 patients. Cardiopulmonary resuscitation was attempted in 454 patients (47%). Collapse of presumed cardiac etiology (337 patients) was either bystander-witnessed (89%), un-witnessed (9%) or EMS personnel-witnessed (2%). Asystole was documented in 55%, ventricular fibrillation or tachycardia in 36% and other non-perfusing rhythms in 9% of these patients. Lay-bystander basic life support was performed in 19%. Nineteen patients (5.6%) survived to hospital discharge and 12 of them were independent in daily life. The survival of subgroups with bystanders-witnessed collapse and bystanders-witnessed ventricular fibrillation was 6.4 and 13%, respectively. Collapse of non-cardiac etiology (117 patients) was preceded by either respiratory failure (41), politrauma (22), circulatory shock (19), cerebrovascular incident (ten), intoxication (nine), strangulation (seven), electrocution (five) or drowning (four patients). Only five patients (4.2%) survived to hospital discharge. Hospital treatment of patients after successful initial cardiopulmonary resuscitation was associated with high mortality and required considerable resources. PMID- 9872639 TI - When an AED meets an ICD... Automated external defibrillator. Implantable cardioverter defibrillator. AB - The chances of prehospital care providers being confronted with a patient with an implantable cardioverter defibrillator (ICD) are increasing and so care providers must receive proper training. Based on observations made during the resuscitation of a patient with an ICD using an automated external defibrillator (AED) some technical features and possible interactions of ICDs and AEDs are highlighted. Furthermore, we discuss the key points of basic knowledge, safety, and treatment protocols for cardiac arrest and other situations required for practical training in the ICD for prehospital care providers. PMID- 9872640 TI - Carbon dioxide narcosis-induced apnea in a rat model of cardiac arrest and resuscitation. AB - In the clinical literature there are reports of patients failing to breathe and becoming comatose when supplied with 100% oxygen for respiratory distress. This effect has been attributed to a loss of respiratory drive. Recent studies have established that this explanation is incorrect, but have left the phenomenon unexplained. We propose that the apnea and coma reported is due to carbon dioxide narcosis. We have reproduced this effect in an animal model and have documented PCO2 values in excess of 250 mmHg during the apneic period. Our results suggest that this level of PCO2 suppresses both brainstem auditory evoked potentials and spontaneous respiration. The high PCO2 is due to inadequate gas exchange, and is easily remedied by provision of adequate ventilation. PMID- 9872642 TI - Removal of a foreign body lodged in the larynx using morphine as a sedative. PMID- 9872641 TI - Effects of respiratory gas composition on transthoracic impedance. AB - The transthoracic current generated during defibrillation comprises a cardiac and extracardiac component. Changes in impedance of transthoracic pathways will alter the transmyocardial current and may thus affect the outcome from defibrillation. The lungs comprise a large component of extracardiac tissue but the effects of different respiratory gases on transthoracic impedance has not been documented. We therefore measured transthoracic impedance (TTI) using different respiratory gas mixtures. TTI across self-adhesive defibrillation pads placed in the antero apical position was measured at end-expiration using a 30 kHz low amplitude AC current. Ten healthy subjects aged 22-34 years (eight male) were studied whilst breathing alternate mixtures of air, 100% oxygen. 70% helium in 30% oxygen, and 70% nitrous oxide in 30% oxygen administered in a random sequence. There was no significant difference in TTI between any of the four respiratory gases. Therefore transthoracic current during defibrillation is unlikely to be affected by different respiratory gases. This is the first study to document that these respiratory gases do not change the impedance of transthoracic pathways. PMID- 9872643 TI - Predicting inhospital cardiac arrest. PMID- 9872644 TI - Comparison of loss heterozygosity in primary and recurrent ductal carcinoma in situ of the breast. AB - Ductal carcinoma in situ (DCIS) of the breast is often an indolent disease, although some cases are reported to recur many years after a limited surgical resection. It is not known whether these recurrences reflect a resurgence of residual disease or an independent development of a second tumor in susceptible individuals. Therefore, we conducted a longitudinal molecular study of four women with reappearance of DCIS 2 to 15 years after an initial conservative resection. Loss of heterozygosity (LOH) was characterized in both tumors in each case, using several polymerase chain reaction-amplified microsatellite markers on five chromosomal arms commonly affected in breast cancer. In three cases with ipsilateral recurrent disease, all of the allelic losses seen in the initial tumors were also seen in the recurrent lesions, suggesting a common genetic pathway for the development of both lesions and continuous proliferation of residual disease. The presence of at least one additional LOH in all of the three recurrent tumors, however, suggests that the recurrent tumors developed after genetic progression. In contrast, in one case of DCIS that was followed by the development of DCIS in the contralateral breast 7 years later (a case of bilateral DCIS), unrelated LOH patterns were present in the two lesions. These findings suggest that the reappearance of DCIS in the same breast is most commonly the result of a tumor derived from (but not identical to) the original lesion, with acquisition of additional genetic changes, even when the recurrent lesion manifested itself many years (15 years, in one case) after the initial presentation. Furthermore, genetic progression could be detected in tumors recurring in as little as 2 years after the initial resection. PMID- 9872646 TI - Expression of cell cycle inhibitor p27 and Ki-67 in human adrenocortical neoplasms. AB - Recent immunohistochemical analysis of cell cycle-related proteins such as p27, a cell cycle inhibitory protein, and Ki-67, a proliferation marker, indicated their possible values in predicting the biologic behavior of various human neoplasms. In this study, we performed an immunohistochemical analysis of p27 and Ki-67 in 42 adrenocortical neoplasms (12 adrenocortical carcinomas, 24 adrenocortical adenomas) and 6 normal adrenal glands to evaluate their possible values in diagnosing adrenocortical malignancy and in predicting the biologic behavior of carcinomas. We detected Ki-67 and p27 immunoreactivity in the nuclei of all of our cases, and we observed a significant negative correlation (r = -0.572, P < .001) between the p27 and Ki-67 labeling indexes (LIs). The LIs of p27 and Ki-67 were 61.7+/-2.6 and 0.28+/-0.08 in the normal adrenal cortex and 59.4+/-6.5 and 0.33+/-0.11 in the adenomas, respectively, with no significant differences between the LIs of the adenomas and normal adrenals. The LIs of p27 and Ki-67 in the carcinomas were 48.9+/-7.5 and 630+/-6.21, respectively. The LI of p27 in the carcinomas was significantly lower than that in the adenomas. The LI of Ki-67 in the carcinomas was significantly higher than that in the adenomas (P < .01). Among carcinoma cases, the Ki-67 LI in living cases tended to be lower than that in deceased cases, and the p27 LI in living cases tended to be higher than that in deceased cases, but these differences did not reach statistical significance. These results indicated that decreased p27 protein expression might cause increased cell proliferation in adrenocortical carcinoma cells in combination with other positive and/or negative regulators of the cell cycle. These results also suggested that immunohistochemical analysis of p27 and Ki-67 might be useful in distinguishing between adrenocortical adenoma and carcinoma PMID- 9872645 TI - A comparison of A103 and inhibin reactivity in adrenal cortical tumors: distinction from hepatocellular carcinoma and renal tumors. AB - Distinguishing adrenal cortical neoplasms from either hepatocellular carcinomas or renal tumors can be difficult. Two recently described antibodies, A103 and inhibin A, are most often reported to be reactive with adrenal cortical neoplasms but with neither hepatocellular carcinoma nor renal cell carcinoma. To compare the sensitivity and specificity of these two antibodies in the diagnosis of adrenal cortical tumors, we stained 22 adrenal cortical adenomas, 4 adrenal cortical carcinomas, 25 hepatocellular carcinomas, and 43 renal tumors, including 33 renal cell carcinomas and 8 oncocytomas, with the A103 and inhibin A using an avidin-biotin complex technique. Fifteen (68%) of 22 adrenal adenomas and 2 (50%) of 4 adrenal cortical carcinomas were reactive with A103. Nineteen (86%) of 22 adrenal adenomas and 3 (75%) of 4 adrenal cortical carcinomas were reactive for inhibin A. None of the renal tumors or hepatocellular carcinomas reacted with A103, but 1 (4%) of 25 hepatocellular carcinomas (a high-grade pleomorphic tumor) and 1 (2%) of 43 renal tumors (a clear-cell renal cell carcinoma) were reactive with inhibin A. The cytoplasmic reactivity for A103 in adrenal tumors was coarsely granular and most common in clear-cell areas. Reactivity for inhibin was either cytoplasmic or membranous and stained both clear-cell and granular areas. We conclude that both antibodies are useful in the immunohistochemical diagnosis of adrenal cortical neoplasms and that A103 is slightly more specific and inhibin slightly more sensitive. PMID- 9872647 TI - Lymphoepithelial cyst and epidermoid cyst of the accessory spleen in the pancreas. AB - We report here two rare cystic lesions, a lymphoepithelial cyst (LEC) and an epidermoid cyst of the accessory spleen (ECAS) occurring in the pancreas. Histologically, the LEC was lined by stratified squamous epithelium and surrounded by a layer of lymphoid tissue with germinal centers. The ECAS showed similar histologic features with scattered lymphoid tissue, but splenic pulp tissue was present in the wall. In both cysts, some small pancreatic islets and ducts were seen in the fibrous tissue surrounding the lymphoid layer or the splenic pulp, respectively. The lining epithelia of the LEC and the ECAS, as well as those of retention cysts of the pancreas and epidermoid cysts of the spleen used for comparison, were similarly positive with AE1/3, CAM5.2, CK7, CK13, and carcinoembryonic antigen. CA19-9 was also detected in the epithelial cells of the LEC, the ECAS, and the retention cyst of the pancreas, but not in those of the splenic epidermoid or branchial cleft cysts used for comparison. These findings indicate that LECs and ECASs might develop from the pancreatic ducts protruding into a lymph node or accessory spleen located in the pancreas, respectively. Some of both cysts might cause elevated levels of serum carcinoembryonic antigen and/or CA19-9 and should be distinguished from malignant cystic tumors. PMID- 9872648 TI - Schwannoma of the pancreas: a report of two cases and a review of the literature. AB - Schwannoma of the pancreas is a rare neoplasm. It generally affects older adults (average age, 60 yr). The sex incidence is equal. These tumors vary considerably in size and approximately two-thirds are partially cystic, which can cause clinical confusion with epithelial neoplasms. Histologically, they are typical schwannomas, with Antoni A and Antoni B areas, and they do not cause undue diagnostic problems once adequate tissue is examined histologically. PMID- 9872649 TI - Immunolocalization of collagen and collagen-binding heat shock protein 47 in fibrotic lung diseases. AB - Pulmonary fibrosis resulting from increased accumulation of various extracellular matrices is a prominent feature in chronic progressive lung diseases. Heat shock protein 47 (HSP47) is a collagen-binding stress protein known to have a specific role in the intracellular processing of procollagen molecules as a collagen specific molecular chaperone in various organs. Possible involvement, however, of HSP47 in relation to increased deposition of collagens in fibrotic lung diseases is not yet known. In this study, we investigated the expression of HSP47 in various pulmonary fibrotic diseases. Formalin-fixed, paraffin-embedded lung sections from 17 autopsies of patients with various pulmonary fibrotic diseases, e.g., organizing pneumonia, interstitial pneumonia, idiopathic pulmonary fibrosis, and diffuse alveolar damage, were stained with monoclonal antibodies for alpha-smooth muscle actin, vimentin, CD68, Type III collagen, and HSP47. The extent of staining was graded semiquantitatively. Five control lung sections were also simultaneously studied. The fibrotic lung sections, in comparison with the control sections, had more interstitial cells positive for alpha-smooth muscle actin and fibroblasts positive for vimentin; we also saw increased infiltration of CD68-positive macrophages. For HSP47, in comparison with the control lung sections, markedly increased immunostaining was always noted in the fibrotic lung sections in association with increased accumulation of Type III collagen in the fibrotic masses. By double immunostaining, colocalization of collagens and HSP47 was noted in the regions of pulmonary fibrosis, and HSP47-expressing cells were found to be mainly alpha-smooth muscle actin-positive interstitial cells. From the above observations, we concluded that overexpression of HSP47 might play an important role in the excessive assembly/synthesis of collagens and could thereby contribute to the fibrosis found in pulmonary fibrotic lung diseases. PMID- 9872650 TI - Expression of ICAM-1, VCAM-1, and LFA-1 in adenocarcinoma of the lung with observations on the expression of these adhesion molecules in non-neoplastic lung tissue. AB - Intercellular adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM 1), and the lymphocyte function-associated antigen (LFA-1) are cell adhesion molecules thought to play an important role in the complex process of airway inflammation and tumor cell growth. The aim of this study was to examine the distribution of ICAM-1, VCAM-1, and LFA-1 in adenocarcinoma of lung and in major cellular compartments of non-neoplastic lung tissue. We examined cellular compartments in tissue from five bronchoalveolar carcinomas, three acinar adenocarcinomas, and one colon cancer metastatic to the lung. The compartments in neoplasms included the tumor cells proper, endothelial cells within the tumor vasculature, tumor stromal cells, and tumor-infiltrating lymphocytes. The compartments in non-neoplastic lung tissue included lung endothelial cells, pulmonary lymphocytes, interstitial fibroblasts, Type II alveolar pneumocytes, and bronchial epithelial cells. ICAM-1 was expressed in tumor cells from all of the nine adenocarcinomas. In contrast, VCAM-1 expression was not identified in tumor cells from any of the nine adenocarcinomas. ICAM-1 was expressed in all cellular compartments of the non-neoplastic lung tissue, whereas VCAM-1 was expressed only in pulmonary lymphocytes and interstitial fibroblastic cells. LFA 1 was uniformly expressed in tumor-infiltrating lymphocytes from each of the nine tumors and all of the lymphocytes in non-neoplastic lung tissue. This study showed major differences in the expression of ICAM-1 and VCAM-1 in tumor cells from pulmonary adenocarcinoma and also provided evidence for a wider distribution of ICAM-1, compared with VCAM-1, in non-neoplastic cellular compartments of the lung. ICAM-1 expression was particularly noticeable in bronchial and alveolar epithelial cells. Upregulation of ICAM-1 in pulmonary adenocarcinoma might foster binding by LFA-1-bearing lymphocytes, with a possible impact on the vulnerability of tumor cells to host defense mechanisms. PMID- 9872651 TI - Sialomucins are characteristically O-acylated in poorly differentiated and colloid prostatic adenocarcinomas. AB - Mucinous glycoproteins are secreted by prostatic adenocarcinomas and might play important roles in tumor invasion and metastasis. Their histochemical properties on routine biopsy specimens have not been fully characterized. We present a histochemical study of mucin in 21 prostatic adenocarcinomas, with particular focus on the demonstration of different types of sialomucins. We applied the following histochemical techniques to routinely processed, formalin-fixed, paraffin-embedded tissue sections: Alcian blue (pH 2.5) and periodic acid-Schiff to reveal both acidic and neutral mucins; high iron diamine and Alcian blue (pH 2.5) to show sulfated and acidic nonsulfated mucosubstances simultaneously; periodic acid borohydride, potassium hydroxide, and periodic acid-Schiff to demonstrate O-acylated sialic acids; periodic acid thionine-Schiff, potassium hydroxide, and periodic acid-Schiff to differentiate pre-existing glycols from those revealed after saponification procedures; and periodic acid borohydride and periodic acid-Schiff to show C9-O-acylated sialic acid. These techniques are useful tools for demonstrating neutral and acidic (sialo- and sulfo-) mucins and di(C8,C9- or C7,C9-)-O-acylated, tri(C7,C8,C9-)-O-acylated and mono(C9)-O acylated sialomucins. Most prostatic adenocarcinomas showed acidic mucins, with sialomucins predominating over sulfomucins. Well-differentiated and moderately differentiated noncolloid tumors had non-O-acylated sialomucins. Poorly differentiated tumors contained mono-O-acylated (C9) sialomucins, and colloid type tumors secreted mono-, di-, and tri-O-acylated sialoglycoproteins. Acidic mucins, mainly sialomucins, constitute the major secretory component in prostatic adenocarcinomas, and our results show that the O-acylation of these sialoglycoproteins inversely correlates with tumor differentiation. Well differentiated and moderately differentiated tumors are not O-acylated, whereas the poorly differentiated ones characteristically have O-acylated sialomucins in C9. Adenocarcinomas of the colloid type, thought to bear a poor prognosis, are the most heavily O-acylated. PMID- 9872652 TI - Increased cell proliferation characterizes Crohn's disease. AB - Patients with long-standing Crohn's disease (CD), a chronic inflammatory intestinal disease, are at increased risk for intestinal cancer. The neoplasia likely results, in part, from deregulated cell proliferation, which allows mutations to become fixed in the crypt progenitor cells. We postulated that tissues derived from patients with CD would exhibit increased mucosal proliferation. Therefore, we examined specimens from 27 consecutive patients with chronic CD with a monoclonal antibody directed against the proliferation marker, Ki-67. The tissues were evaluated histologically, and the Ki-67 immunostaining patterns were recorded. The antibody to Ki-67 stained the bases of the crypts in both the small and large intestines. The mean number of Ki-67 immunoreactive cells in the normal crypt was 34.1 versus 95.1 in the regenerative mucosa and O in areas of pyloric metaplasia (P < .00001). Ki-67 staining of the mucosa of patients with CD confirmed that cell proliferation is markedly increased and that the replicating compartment of each crypt during regeneration is expanded. We concluded that the increased cell proliferation might predispose the mucosa to mutational events, thereby increasing the cancer risk in these patients. The lack of proliferation in areas of pyloric metaplasia might represent a mucosal adaptive response of the lower crypt that decreases the number of cycling cells vulnerable to genetic damage. Furthermore, growth factors produced by these cells might promote healing of the damaged mucosa. PMID- 9872653 TI - CD45 (leukocyte common antigen) immunoreactivity in metastatic undifferentiated and neuroendocrine carcinoma: a potential diagnostic pitfall. AB - Leukocyte common antigen (CD45/LCA) and keratin expression are generally mutually exclusive in diagnostic surgical pathology. CD45 reactivity is a reliable indicator of the hematolymphoid nature of a tumor, whereas keratin reactivity is typical of epithelial differentiation (carcinomas and some sarcomas). Some lymphomas, however, might lack detectable CD45 expression, whereas occasional ones might express keratins. CD45 immunoreactivity has been considered exquisitely specific for hematopoietic cells. We report three undifferentiated or neuroendocrine carcinomas that showed membrane-associated immunoreactivity for CD45 in addition to showing distinctive keratin cocktail (AE1/AE3) and epithelial membrane antigen reactivity (all cases); also, keratin 7 was demonstrated in one case and keratin 19 in another. Two cases were lymph node metastases of undifferentiated carcinomas, one of them from the lungs and the other of an unknown origin; the former case showed neuroendocrine features. The third case represented a pulmonary large-cell undifferentiated carcinoma. These cases were negative for lineage-specific leukocyte antigens and did not show clonal immunoglobulin heavy-chain gene rearrangements. Electron microscopic studies demonstrated desmosomes and keratin-like tonofilaments in all three cases, thus confirming the epithelial nature of these tumors. The exceptional membrane staining for CD45 seen in these undifferentiated carcinomas might be comparable to experimentally detected incorporation of leukocyte antigens into the cell membranes of nonleukocytic cells in a leukocyte-rich environment. This rare diagnostic pitfall should be considered in the diagnostic surgical pathology of undifferentiated tumors. It is best avoided by employing a panel of leukocyte and epithelial antigens and by use of electron microscopy, if possible. PMID- 9872654 TI - The immunophenotype of blast transformation of chronic myelogenous leukemia: a high frequency of mixed lineage phenotype in "lymphoid" blasts and A comparison of morphologic, immunophenotypic, and molecular findings. AB - Immunophenotypic studies have a limited role in the diagnosis of chronic myelogenous leukemia (CML) but are increasingly being used in CML blast transformation (BT). Determination of the cell lineage of CML blasts is clinically important because patients with lymphoid blast transformation have a better response to chemotherapy and longer survival than those with other lineages. We studied the morphologic, cytochemical, immunophenotypic, cytogenetic, and molecular features of 20 patients with Philadelphia chromosome positive CML and more than 10% blast cells in peripheral blood or bone marrow. The blasts were morphologically heterogeneous. CD33 was expressed in 19 cases (95%), followed by CD13 (85%), CD11c (80%), CD36 (60%), CD117 (40%), and CD15 (30%). Seven cases (35%) had a precursor-B lymphoid immunophenotype, and 13 (65%) had a predominantly myeloid immunophenotype. Of the former group, of which only one had a pure lymphoid phenotype, terminal deoxynucleotidyl transferase (TdT) and CD19 were expressed in 100%, CD10 in 85.7%, and CD20 in 14.3%. Of the latter group, all 13 expressed from 3 to 6 myeloid antigens, with 46.2% myeloperoxidase positive and 69.2% CD61 positive. No cases were interpreted as T lineage, but the T-cell antigens CD3, CD4, CD5, and CD7 were expressed in 5.0, 40.0, 5.3. and 30.0% of all cases, respectively. In most cases, the immunophenotype of the CML blasts could not be predicted from their morphologic features. Polymerase chain reaction showed that 80.0% of the lymphoid group and 37.5% of the myeloid group had immunoglobulin heavy-chain gene rearrangements. The frequent lineage infidelity of the blast cells in CML BT seems to be related to the stem cell origin of this disorder. Such lineage infidelity, however, makes classification of many cases difficult and the significance of and criteria for biphenotypic blast crisis of CML is yet to be determined. PMID- 9872655 TI - N-myc gene amplification in rhabdomyosarcoma detected by fluorescence in situ hybridization: its correlation with histologic features. AB - Fluorescence in situ hybridization (FISH) was applied to 15 alveolar rhabdomyosarcomas (A-RMSs) and 14 embryonal RMSs (E-RMSs) to detect N-myc (also called MYCN) oncogene amplification. The results were compared with histologic characteristics and clinical factors. The number of surviving patients in each subtype was 5 of 15 with A-RMS and 5 of 14 with E-RMS. N-myc amplification was detected in 9 of the 15 A-RMSs but in none of the 14 E-RMSs. Tumor cells exhibiting N-myc amplification were identified only in the alveolar area in two A RMSs, and they demonstrated a histologic mixture of alveolar and embryonal patterns. The remaining seven cases with an amplified N-myc showed a conventional alveolar pattern. Among the 15 A-RMSs, the survival rate of patients with tumors showing nonamplified N-myc and amplified N-myc oncogene was 4 (66%) of 6 and 1(11%) of 9, respectively (P < .05). No significant difference was observed between the other clinical findings (age, primary sites, clinical stages) of the N-myc-amplified and nonamplified tumors. Therefore, we concluded that the N-myc gene amplification, which is characteristic of a particular subtype of A-RMS, might be useful as a prognostic factor for an unfavorable outcome. PMID- 9872656 TI - Establishment and molecular characterization of five cell lines derived from renal and extrarenal malignant rhabdoid tumors. AB - Malignant rhabdoid tumor (MRT) is a rare, enigmatic childhood cancer characterized by extreme aggressiveness and resistance to chemotherapy. To understand better the origin of the tumor and the mechanisms by which it develops and resists treatment, five cell lines were established from patients presenting with MRT (two renal and three extrarenal tumors). All of the cell lines display the light microscopic and ultrastructural features, as well as the variable immunohistochemical profile, characteristic of MRT. All are capable of forming tumors in nude mice. Three of the cell lines have detectable abnormalities of chromosome 22: one a t(22, 22) unbalanced translocation and two others a loss of heterozygosity of polymerase chain reaction-based microsatellite markers. Northern blot analysis showed that overexpression of the c-myc message was a consistent characteristic of the five MRTs evaluated. Although mutations of the p53 gene were not detectable by sequence analysis, all of the cell lines showed nuclear accumulation of the p53 protein by an immunocytochemical analysis in a minority of the cells. This result suggests that dysfunction in a p53-dependent apoptotic pathway might play a role in the multiple drug resistance phenotype of these tumors. PMID- 9872658 TI - Quantification of tissue eosinophils and lymphocytes in histologic sections. AB - During a study of eosinophil-predominant gallbladder disease, an image analysis (IA) technique was developed for quantification of eosinophils and lymphocytes in routine formalin-fixed tissue sections. Alternating sections were stained with hematoxylin and eosin for eosinophils and with monoclonal CD45 antibody visualized with diaminobenzidine by an avidin-biotin procedure for lymphocytes. A protocol was then developed using a commercially available image analyzer and two well-defined macro routines. The system was validated with cell block sections prepared from peripheral blood samples with known eosinophil and lymphocyte counts. The eosinophil counts obtained by this IA technique showed excellent correlation with the absolute counts from a peripheral blood analyzer (r2 = 0.987). The lymphocyte counts obtained by IA showed good correlation with the absolute counts (r2 = 0.820). This IA-based technique provides a sensitive, reproducible, and substantiated means of quantifying inflammatory cells in tissue sections. This rapid and easily learned technique adds quantification as a complementary dimension to the subjective assessment of tissue morphology. PMID- 9872657 TI - In vivo cyclin E expression as a marker for early cervical neoplasia. AB - Identification of human papillomavirus (HPV)-related early cervical neoplasia and its distinction from benign epithelial alterations is based on either subjectively applied morphologic criteria or on identification of associated papillomaviruses. The direct and indirect consequences of HPV infection, however, potentially include upregulation of some host genes. We investigated one candidate, cyclin E, as a possible marker for HPV-related early squamous lesions. Serial paraffin sections from 92 archival cervical biopsy specimens were analyzed, including 19 non-neoplastic biopsy specimens, 30 low-grade and 31 high grade squamous intraepithelial lesions (SILs), and 12 invasive carcinomas. Four parameters (histologic diagnosis, cyclin E staining, HPV status and, in selected cases, Ki-67 staining) were scored, and their relationship(s) were evaluated by the chi2 independence test. Twenty-one, 73, 79, and 75% of nonlesional epithelia, low-grade SILs, high-grade SILs, and invasive squamous cell carcinomas, respectively, were HPV positive (P < .001 for HPV status vs. diagnosis). Cyclin E staining was nuclear in distribution, and the frequency of positive staining, ie., moderate or strong intensity, was significantly higher (P < .001 for cyclin E staining vs. diagnosis) in all of the lesional epithelia (92.3, 51.6, and 50% of low-grade and high-grade SILs and carcinomas, respectively) compared with nonlesional epithelium (5.9%). Cyclin E positivity and/or HPV positivity was seen in 100% of the low-grade SILs. Epithelial Ki-67 and cyclin E staining were strikingly different in frequency and distribution. Cyclin E was undetectable in basal cells of normal mucosa (which were positive for Ki-67) and limited to suprabasal epithelium in noninvasive lesions. Cyclin E expression correlates strongly with morphologic features of HPV-related preinvasive and invasive cervical disease. This correlation is most pronounced in low-grade SILs. The possibility that in vivo cyclin E staining is a generic marker for HPV infection in low-grade SILs merits additional study. PMID- 9872659 TI - Lymphoepithelioma-like carcinoma of the renal pelvis: a case report with immunohistochemical analysis and in situ hybridization for the Epstein-Barr viral genome. AB - Lymphoepithelial carcinoma is a relatively common malignancy in the nasopharynx, but it rarely occurs at other sites. Described herein is the first case of a renal pelvic neoplasm that closely resembled lymphoepithelial carcinoma, with analyses of histology, immunophenotype, in situ hybridization for the Epstein Barr viral genome, and flow cytometric DNA ploidy. The tumor was detected in an 70-year-old Japanese man who presented with hematuria Histologic examination showed an undifferentiated round or spindle cell carcinoma (cytokeratin 7+, cytokeratin 20+, epithelial membrane antigen+, vimentin-) with abundant lymphocytes (predominantly UCHL-1+ T cells), plasma cells, and macrophages in and around the tumor cell nests. The tumor was limited to the pelvis, with a minute focus of carcinoma in situ. No Epstein-Barr viral genomic sequences were detected by in situ hybridization. The tumor had an aneuploid DNA content. The patient remains well without disease 6 years after surgery and radiotherapy. Recognition of this type of renal pelvis carcinoma is important to avoid misdiagnosis. PMID- 9872660 TI - Recommendations for the reporting of soft tissue sarcomas. Association of Directors of Anatomic and Surgical Pathology. PMID- 9872661 TI - Hot off the screen. PMID- 9872662 TI - Meiosis of wheat x lymegrass hybrids. AB - Meiosis was examined in pollen mother cells of F1 hybrids made from crosses between wheat (Triticum aestivum) and lymegrass (Leymus arenarius and L. mollis). Fluorescence genomic in situ hybridization detected pairing between wheat and lymegrass chromosomes during prophase I and metaphase I. Such pairing, when resulting in bivalent formation, was likely to yield correct disjunction, and hence intergenomic recombination could be incorporated into the gametes. Bivalents in these hybrids, however, were more frequently formed between chromosomes of the same parental origin. Univalents were common, whereas multivalents were not clearly detected. Meiotic behaviour in some cells was not totally aberrant, and this may have accounted for the presence of normal pollen. The results are discussed in relation to intergenomic pairing, meiotic behaviour in wide-hybrids and genome relationships, including the Leymus genome origin. PMID- 9872663 TI - Multiple locations of the rDNA sites in holocentric chromosomes of Rhynchospora (Cyperaceae). AB - Several cytogenetic studies have shown that representatives of the family Cyperaceae have holocentric chromosomes. Despite their interesting chromosome morphology, the chromosome organization has not been studied. This paper reports on the number and distribution of 18S-5.8S-26S ribosomal RNA sites by fluorescence in situ hybridization in eight Brazilian species of Rhynchospora. The signal of the rDNA probe was always localized in the telomeric regions. A high degree of variation was observed in the number of labelled sites, ranging from 4-8 in karyotypes with 2n = 10 to 30 sites in a karyotype with 50 chromosomes. It is possible that the same mechanism involved in the multiplication of these regions in organisms with monocentric chromosomes also plays a role in the polymorphism observed in holocentric chromosomes of Rhynchospora. An interesting feature of most hybridization sites was their diffuse state observed through to early metaphase. The decondensed state probably reflects the later transcription of this region during the cell cycle. PMID- 9872665 TI - Characterization of reciprocal translocations in pigs using dual-colour chromosome painting and primed in situ DNA labelling. AB - We report the use of dual-colour chromosome painting to determine the exact nature of certain chromosome rearrangements observed in the pig (Sus scrofa domestica). The chromosomal abnormalities were detected by GTG- and RBG-banding techniques. The initially proposed interpretations were: (1) rcp(6;13)(p1.5;q4.1); (2) rcp(11;16)(p1.4;q1.4); (3) rcp(6;16)(p1.1;q1.1); (4) rcp(13;17)(q4.1;q1.1); (5) rcp(6;14)(q2.7;q2.1); (6) rcp(3;5)(p1.3;q2.3); (7) rcp(2; 14)(q1.3;q2.7); (8) rcp(15;17)(q1.3;q2.1). Hybridizations were carried out with biotin- and digoxigenin-labelled probes obtained by priming authorizing random mismatches polymerase chain reaction (PARM-PCR) amplification of porcine flow-sorted chromosomes. In some cases, i.e. (1), (4), (5), (6), (7) and (8), the fluorescence in situ hybridization (FISH) results allowed confirmation of the interpretations proposed with classical cytogenetic methods. Chromosome painting proved the reciprocity of the translocation in cases (1), (6) and (8), whereas modifications of the formula were proposed for case (2). Primed in situ DNA labelling (PRINS) experiments have also been carried out in case (3) using a primer specific for the centromeres of acrocentric chromosomes (first experiment) or a primer specific for the centromeres of a subset of meta- and submetacentric chromosomes including chromosome 6 (second experiment). It allowed us to demonstrate that the breakpoints occurred in the centromeric region of chromosome 16 and in the p. arm of chromosome 6, just above the centromere. PMID- 9872664 TI - Repetitive DNA sequences in the common vole: cloning, characterization and chromosome localization of two novel complex repeats MS3 and MS4 from the genome of the East European vole Microtus rossiaemeridionalis. AB - We have characterized two novel, complex, heterochromatic repeat sequences, MS3 and MS4, isolated from Microtus rossiaemeridionalis genomic DNA. Sequence analysis indicates that both repeats consist of unique sequences interrupted by repeat elements of different origin and can be classified as long complex repeat units (LCRUs). A unique feature of both repeat units is the presence of short interspersed repeat elements (SINEs), which are usually characteristic of the euchromatic part of the genome. Comparative analysis revealed no significant stretches of homology in the nucleotide sequences between the two repeats, suggesting that the repeats originated independently during the course of vole genome evolution. Fluorescence in situ hybridization analysis demonstrates that MS3 and MS4 occupy distinct domains in the heterochromatic regions of the sex chromosomes in M. transcaspicus and M. arvalis but collocalize in M. rossiaemeridionalis and M. kirgisorum heterochromatic blocks. The localization pattern of the repeats on the vole chromosomes confirms the independent origin of the two repeats and suggests that expansion of the heterochromatic blocks has occurred subsequent to speciation. PMID- 9872666 TI - Torsional state of DNA in a transcriptionally hyperactive Balbiani ring of polytene chromosomes. AB - The torsional tension of unconstrained double-helical DNA was determined in transcriptionally hyperactive Balbiani ring 2 (BR2) and in inactive polytene chromosome bands of Chironomus tentans. The method used is based on the dual ability of small intercalating ligands to (a) sense, by differential binding, twists that deviate from that of regular B-form DNA and (b) create positive torsional tension in closed double-stranded DNA, thereby compensating for any negative torsional tension that existed before intercalation. Isolated nuclei of salivary glands were stained with the intercalating fluorescent dye ethidium bromide (EtBr) at various concentrations, and the temporal fluorescence intensity changes (deltaI/I per min) occurring in BR2 and in inactive bands were monitored under a confocal laser scanning microscope during the process of DNA nicking by laser irradiation or DNAase I. From the EtBr concentration at which deltaI/I per min was neither positive nor negative after nicking (i.e. at the equivalence point), the relative twist difference (RTD) was calculated. In bands, it was found to be very small, suggesting that their unconstrained DNA is under low torsional stress. In contrast, the RTD of DNA in highly expanded areas of BR2 was estimated to be negative and of a significant magnitude in absolute terms. This indicates that transcriptionally hyperactive DNA is under considerable negative torsional tension. PMID- 9872667 TI - XY sex reversal in the wood lemming is associated with deletion of Xp21-23 as revealed by chromosome microdissection and fluorescence in situ hybridization. AB - In the wood lemming (Myopus schisticolor), XY sex reversal occurs naturally because of the presence of an X chromosome variant designated X*. The two types of X chromosome, X and X*, can be distinguished by G-banding, and analyses have demonstrated complex rearrangements of the short arm of X*. Here, chromosomal microdissection, degenerate oligonucleotide-primed polymerase chain reaction (DOP PCR) and fluorescence in situ hybridization (FISH) techniques have been used to generate and map DNA probes for different parts of the X and X* chromosomes. The results showed that the region of Xp21-23 is deleted from the X* and some of the deleted DNA sequences are homologous to the mouse gamma-satellite. The deletion must be associated with the sex reversal in this species. FISH experiments with dissected probes of X and distal half of Xq provided evidence for presence of homologous sequences between large regions of the X and Y chromosomes, including euchromatic and heterochromatic parts of the sex chromosomes. The findings of this study will be of significance for further cloning of important candidate gene(s) responsible for the XY sex reversal. PMID- 9872668 TI - Pattern of chromosomal localization of the Hoppel transposable element family in the Drosophila melanogaster subgroup. AB - We have isolated a Hoppel-like transposon from heterochromatin of the second chromosome of Drosophila melanogaster and used a conserved DNA sequence between the different elements of this family to determine their distribution in both mitotic and polytene chromosomes. The hybridization pattern of polytene chromosomes extends throughout the entire chromocentre, as well as a substantial portion of the fourth chromosome. Analysis of different wild-type strains of D. melanogaster shows variation in euchromatic insertion sites, although most insertions are found near the chromocentre. The positions and the number of heterochromatic clusters of Hoppel on mitotic chromosomes are conserved among the several strains analysed. Accurate mapping of this element was achieved by in situ hybridization on D. melanogaster mitotic chromosomes that had previously been banded with Hoechst 33258. To evaluate the evolutionary stability of this pattern, different species were analysed by in situ hybridization and Southern blotting. We conclude that Hoppel has a conserved distribution in mitotic heterochromatin within the D. melanogaster subgroup, established around 5 million years ago. The overall conservation of heterochormatic organization supports the notion that heterochormatin does perform important structural and functional roles. PMID- 9872671 TI - A rare chromosome 5 heterochromatic variant derived from insertion of 9qh satellite 3 sequences. AB - A rare chromosome 5 heterochromatic variant not linked to any clinical sign was identified in a three-generation family. After performing conventional cytogenetics characterization, fluorescence in situ hybridization of D9Z1 indicated that the unusually large qh region of chromosome 5 originated from 9qh, whereas the centromere of the variant chromosome was 5-specific as demonstrated by primed in situ DNA labelling. FISH of probes targeting satellite 3 and beta satellite sequences of 9qh showed that only satellite 3 sequences were present in the variant 5qh region. PMID- 9872670 TI - Electron microscopic in situ hybridization of digoxigenin-dUTP-labelled DNA probes with Drosophila melanogaster polytene chromosomes. AB - We report a simplified method of electron microscopic (EM) in situ hybridization for standard squashes of Drosophila melanogaster polytene chromosomes using digoxigenin-11-dUTP labelled DNA probes. The method is efficient and reproducible: its high resolution and specificity were demonstrated for the transformed strain 148, in which the insertion was localized precisely as a new thin band both by conventional EM and according to our method. In addition, the method was applied to the fine mapping of the developmentally regulated gene muscle-blind (mbl). On the one hand, mbl was shown to cover the 54B1-2 large band and the adjacent interbands in the 2R polytene chromosome. On the other hand, the use of distantly located DNA probes in the mbl gene allowed us to orientate the transcription unit in the chromosome. PMID- 9872669 TI - Comparative methylation analysis of murine transgenes that undergo or escape X chromosome inactivation. AB - We analyzed an X-linked metallothionein-vasopressin (MTVP) fusion transgene that undergoes X-chromosome inactivation (X inactivation) and an X-linked transferrin (TFN) transgene that escapes X inactivation with respect to methylation in the 5' regulatory regions. The MTVP transgene promoter region is unmethylated when the transgene is on the active X chromosome and methylated when on the inactive X chromosome. Interestingly, the MTVP transgene is not detectably transcribed from the male X chromosome, although it is unmethylated, consistent with its availability for transcription. The TFN transgene promoter region is hypomethylated on both the active and inactive X chromosomes, consistent with its expression from both chromosomes. The TFN and MTVP transgenes have been mapped to chromosomal regions D and C, respectively, by fluorescence in situ hybridization. These observations are discussed in the context of our understanding of the role of DNA methylation in the spread and maintenance of X-chromosome inactivation. PMID- 9872673 TI - T-cell receptor TCRG1 and TCRG2 clusters map separately in two different regions of sheep chromosome 4. PMID- 9872672 TI - An 84-kilobase physical map and repeat polymorphisms of the gastrin/cholecystokinin brain receptor region at the junction of chromosome segments 11p15.4 and 15.5. PMID- 9872674 TI - Dolphin beta-glucocerebrosidase, map position 1q22. PMID- 9872676 TI - Interleukin-10 stabilizes inhibitory kappaB-alpha in human monocytes. AB - Interleukin-10 (IL-10) protects animals from lethal endotoxemia. This beneficial effect is mediated, in part, by inhibition of inflammatory cytokine production, including tumor necrosis factor-alpha (TNF-alpha). Evidence suggests that IL-10 may inhibit activation of the transcription factor nuclear factor-kappaB (NF kappaB) through an unknown mechanism. NF-kappaB activation in response to inflammatory signals is dependent upon degradation of its associated inhibitory peptide, inhibitory kappaB-alpha (IkappaB-alpha). We hypothesized that IL-10 prevents human monocyte NF-kappaB activation and resultant TNF-alpha production by stabilization of IkappaB-alpha. The purpose of this study was to determine the effect of IL-10 on lipopolysaccharide (LPS)-induced human monocyte TNF-alpha production, NF-kappaB activation, and IkappaB-alpha degradation. Monocytes were isolated from human donors. Cells were stimulated with endotoxin (LPS, 100 ng/mL) with and without human IL-10 (10 ng/mL). Following stimulation, TNF-alpha was measured in cell supernatants by ELISA, NF-kappaB activity by electrophoretic mobility shift assay, and IkappaB-alpha levels by Western blot. We observed that after LPS stimulation of human monocytes, TNF-alpha increased to 798+/-67 pg/mL (p < .001 versus control). IL-10 attenuated LPS-stimulated TNF-alpha production (297+/-54; p < .001 versus LPS alone). After LPS stimulation in human monocytes, IkappaB-alpha protein levels decreased, and NF-kappaB DNA binding increased. IL 10 pretreatment prevented LPS-induced decreases in IkappaB-alpha protein levels and attenuated NF-kappaB DNA binding. IL-10 appears to prevent activation of NF kappaB by preserving IkappaB-alpha protein levels, leading to a reduction in TNF alpha release. PMID- 9872677 TI - Hypophysectomy, high tumor necrosis factor levels, and hemoglobinemia in lethal endotoxemic shock. AB - Experimental models of lethal endotoxemia in rodents are widely used to delineate pathogenic mechanisms of inflammation, sepsis, and septic shock. One long standing but poorly understood observation is that removal of the pituitary gland (hypophysectomy) renders experimental animals 1,000-fold more sensitive to the lethal sequelae of lipopolysaccharide (LPS). Previous explanations for this phenomenon focused on hypophysectomy-induced deficiencies of corticosteroids, because glucocorticoids effectively suppress the synthesis of tumor necrosis factor (TNF), which is a primary mediator of LPS lethality. We measured LPS stimulated macrophage TNF release in the presence of serum from hypophysectomized rats to detect the appearance of an inducible 65 kDa protein that enhances TNF release. Surprisingly, the N-terminal amino acid sequence analysis of the isolated, purified protein revealed its identity as hemoglobin. Hypophysectomy significantly increases serum hemoglobin levels (control hemoglobin = 103+/-18 microg/mL versus hypophysectomized serum hemoglobin = 279+/-13 microg/mL; p < .05). Purified hemoglobin enhances TNF synthesis in LPS-stimulated macrophages by at least 1,000-fold, which is specifically inhibited by antihemoglobin antibodies. Thus, hemoglobin mediates increased TNF synthesis in endotoxemic, hypophysectomized rats. This mechanism of increased TNF release has potential implications for patients with hemoglobinemia following blood transfusion, surgery, injury, infection, or other conditions that can be associated with endotoxemia and sepsis. PMID- 9872678 TI - Role of NO and endothelin in hemoglobin-induced pulmonary vasoconstriction. AB - The underlying mechanisms of hemoglobin (Hb)-induced vasoconstriction are not yet well understood. The aim of this study was to elucidate the influence of nitric oxide (NO) and endothelin (ET) on Hb-induced pulmonary vasoconstriction. Therefore, an autologous Hb preparation was administered into isolated rabbit lungs, in which pulmonary artery pressure (PAP) and weight gain was monitored. Either glyceroltrinitrate (GTN; 10(-5) M; n=6), L-arginine (10(-2) M; n=6), L NAME (10(-4)M; n=6), ET(A)- or ET(B)-receptor antagonists (BQ,23, 10 6M, n=6) or (BQ788, 10(-6) M, n=6) were added to the perfusion fluid and NOx and thromboxane A2 levels were measured. RESULTS: In the control group the Hb-stimulation resulted in a pressure response up to 25.1+/-2.1 mmHg (p < .05), which was 136+/ 6% of the reference value. The PAP increase was significantly (p < .05) blunted after GTN (71+/-5%), L-arginine (93+/-6%) and BQ788 (88+/-7%). Pretreatment with L-NAME (139+/-13%) or BQ123 (115+/-9%) did not show significant changes in PAP. CONCLUSION: The reduction of the Hb-induced pulmonary hypertension by NO-donors points toward the inactivation of NO by free hemoglobin. Likewise, ET(B)-receptor mediated vasoconstrictive effects without changes in NOx concentrations seem to play a pathogenetic role in the Hb-induced pulmonary vasoconstriction. PMID- 9872679 TI - Post-hemorrhagic shock mesenteric lymph is cytotoxic to endothelial cells and activates neutrophils. AB - The goal of these experiments was to test the hypothesis that after a nonlethal episode of hemorrhagic shock, factors carried in the mesenteric lymph would promote endothelial cell injury and activate neutrophils to a greater extent than portal vein plasma. Catheters were placed in the efferent lymphatic duct draining the mesenteric lymph node complex, after which male rats were subjected to sham or actual shock (30 mmHg for 90 min), and lymph was collected. Portal vein plasma was collected from the sham-shock and shocked rats at 6 h post-shock or sham shock. When the effect of lymph or portal blood plasma was tested on endothelial cell (HUVEC) monolayer permeability, it was found that post-shock lymph, but not post-shock portal vein plasma, increased HUVEC permeability to both 10 kDa and 40 kDa permeability probes. Subsequent experiments documented that only post-shock lymph was cytotoxic to endothelial cells as manifest both by decreased trypan blue dye exclusion and the increased release of Chromium-51 from chromium-loaded endothelial cells. Furthermore post-shock lymph induced a greater increase in neutrophil superoxide formation than pre-shock lymph, pre-shock, or post-shock portal vein plasma. Lastly, neutrophil-mediated endothelial cell injury was potentiated by the presence of post-shock lymph, and the magnitude of HUVEC injury was greater in endothelial cells incubated with post-shock lymph plus neutrophils than in monolayers incubated with post-shock lymph or neutrophils alone. These results suggest that post-shock lymph is cytotoxic to endothelial cells and activates neutrophils. Since the lung is the first organ that is exposed to mesenteric lymph, lung injury after hemorrhagic shock may be mediated by factors contained in mesenteric lymph. PMID- 9872680 TI - Mesenteric lymph: the critical bridge between dysfunctional gut and multiple organ failure. PMID- 9872681 TI - Selective microvascular endothelial cell dysfunction in the small intestine following resuscitated hemorrhagic shock. AB - Following resuscitation (RES) from hemorrhagic shock (HEM), intestinal microvessels develop progressive vasoconstriction that impairs mucosal blood flow, despite central hemodynamic RES. These events might have clinical consequences secondary to occult intestinal ischemia. We hypothesized that the microvascular impairments were due to progressive endothelial cell dysfunction and an associated reduction in the dilator, nitric oxide (NO), following HEM/RES. Male Sprague-Dawley rats, were monitored for central hemodynamics and the terminal ileum was studied with in vivo videomicroscopy. HEM was 50% of baseline mean arterial pressure (MAP) for 60 min, and RES was with shed blood + 1 volume of normal saline (NS). Following HEM/RES, acetylcholine (10)(-7), 10(-5) M) was topically applied and ileal inflow (A1) and premucosal arteriolar diameters were measured to assess endothelial-cell function at 60 and 120 min post-RES. Normalization of MAP, cardiac output, and heart rate demonstrated adequate systemic resuscitation. Post-RES vasoconstriction developed in A1 (-25%) and premucosal (-28%) arterioles with an associated reduction in A1 flow (-47%). However, there was a selective impairment of endothelial-dependent dilation that was manifested only in the smaller premucosal arterioles and not in the inflow, A1 arterioles. This suggests that multiple mechanisms are involved in the development of the post-RES vasoconstriction. The premucosal response was likely mediated by endothelial cell dysfunction, while the A1 response was probably the result of enhanced vasoconstrictor forces. This early microvascular dysfunction might contribute to the late sequelae of intestinal ischemia and might alter microvascular responses to subsequent systemic insults. PMID- 9872683 TI - Aminothiol WR-1065 protects endothelial cell morphology against alterations induced by lipopolysaccharide. AB - In septic patients, lipopolysaccharide (LPS) damages the vascular endothelium, which manifests as tissue edema and impaired healing. This pathology occurs when LPS distorts endothelial cell morphology partly by generating free radicals. A radioprotector that scavenges free radicals, the aminothiol WR-1065 ([N-2 mercaptoethyl]-1-3-diaminopropane) was found in a prior study to normalize the morphology of irradiated endothelial cells (Mooteri SN, Podolski JL, Drab EA, et al: Radiat Res 145:217-224, 1996). The aim of this study was to determine whether WR-1065 also normalized endothelial cell morphology following exposure to LPS. For this aim, portions of bovine aortic endothelial cell cultures were denuded and exposed to LPS at 1 ng/mL. After 30 min, the apical membrane expressed increased integrin receptor to fibronectin, alpha5beta1. After 5 h, the morphology of the cells at the leading edge was distorted, and cell-cell contact was lessened. Also, filamentous actin-containing stress fibers were dissipated; however, filamentous actin content per cell was unchanged. Treatment with 2 mM WR 1065 for 2 h prior to LPS exposure attenuated the increased expression of alpha5beta1 and promoted cell-cell contact in the migrating endothelial cells. WR 1065 also promoted the retention of stress fibers and actin cytoskeletal shape in cells treated with LPS. Thus, LPS distorted endothelial cell morphology after increasing apical membrane expression of alpha5beta1 and dissipating stress fibers, effects prevented by WR-1065. PMID- 9872682 TI - Aminothiols protect endothelial cell proliferation against inhibition by lipopolysaccharide. AB - Lipopolysaccharide (LPS) is a primary agent of sepsis that damages the vascular endothelium. Endothelial cell proliferation is key to the repair of damaged endothelium, and drugs that counteract the antiproliferative impact of LPS on endothelial cells should be beneficial. Because LPS exerts much of its cytotoxicity by generating reactive oxygen and nitrogen intermediates, it would be helpful to know whether therapeutic antioxidant thiols maintain cell proliferation in injured endothelium. In this study, it was found that LPS inhibited bovine aortic endothelial cell proliferation by inducing apoptosis and by decreasing DNA synthesis. Because of its benefit to irradiated endothelial cells, we then treated the cells with a radio- and chemoprotective aminothiol, WR 1065 ([N-2-mecaptoethyl]-1-3-diaminopropane, the active form of Amifostine/Ethyol). WR-1065 attenuated the inhibition of DNA synthesis caused by LPS exposure. The disulfide of WR-1065, WR-33278, was tested and shown to both promote DNA synthesis and inhibit apoptosis. The effectiveness of the disulfide suggests that the reduction of cytotoxicity does not necessarily result from the scavenging of free radicals. These findings demonstrate a novel role for aminothiols in promoting DNA synthesis and lowering apoptosis in endothelium injured with LPS. PMID- 9872685 TI - Animal models of sepsis and shock: a review and lessons learned. Edwin A Deitch. Shock 9(1):1-11, 1998. PMID- 9872684 TI - Acetazolamide treatment prevents in vitro endotoxin-stimulated tumor necrosis factor release in mouse macrophages. AB - We previously showed that incubation in carbon dioxide (CO2), but not air or helium (He), markedly decreased macrophage intracellular pH (pHi) and resulted in reversible inhibition of lipopolysaccharide- (LPS) stimulated tumor necrosis factor (TNF) and interleukin-1 release. We sought to determine whether carbonic anhydrase inhibition with acetazolamide would prevent CO2-mediated inhibition of LPS-stimulated TNF release. Murine peritoneal macrophages were treated with acetazolamide for 1 h under control atmosphere (95% air/5% CO2) and then switched to incubator modules containing: 1) 80% CO2/20% O2, 2) 80% He/20% O2, or 3) 100% air. Before transfer to experimental atmospheric conditions the macrophages were stimulated with 0 or 1 microg/mL of LPS (Escherichia coli 0111 B4). Supernatant TNF was measured 4 h later by bioassay. In parallel experiments LPS-stimulated cytokine mRNA was estimated using reverse transcriptase polymerase chain reaction (RT-PCR) 2 h after LPS stimulation. Viability was determined using dye uptake. Incubation in CO2 or helium had no effect on TNF production in the absence of LPS. In the absence of acetazolamide CO2 produced marked inhibition of LPS stimulated TNF release, but this was not blocked by the presence of acetazolamide. This CO2-mediated inhibition of TNF was associated with normal levels of TNF mRNA. In acetazolamide-treated macrophages, LPS resulted in a dose dependent inhibition of TNF release when the cells were incubated in air or helium. Maintenance of normal intracellular pH is required for TNF release, but not TNF mRNA induction by LPS. Factors that alter intracellular pH regulation may modulate LPS-stimulated cytokine production. PMID- 9872686 TI - A cost-effectiveness model for analyzing two varicella vaccination strategies. AB - A model for estimating the cost-effectiveness of a program for vaccinating employees at a health care institution against varicella zoster virus (VZV) was studied. Three outcomes of varicella vaccination--cost-effectiveness, reduction in employee infections, and reduction in patient exposures--were stratified to estimate the incremental costs of vaccinating three employee groups. The groups consisted of all employees (vaccinate-all group), employees providing direct patient care (direct care group), and employees working in high-risk patient care areas (high-risk group). Two strategies for employee vaccination were used: screen (by antibody titer testing) and then vaccinate, and vaccination alone. A model was used to estimate the various outcomes of the vaccination program. The model supported the screen, then vaccinate strategy of vaccinating employees, at a cost savings of about $50 per employee. Vaccination of all employees prevented 35 employee infections and 674 patient exposures for every 10,000 potentially susceptible employees. The cost of preventing one employee infection was about $15,000, and the cost of preventing one patient exposure was about $775. An employee vaccination program is a good investment in preventing patient exposures to VZV and may be cost-effective compared with only screening employees. PMID- 9872687 TI - Using multiple pharmacoeconomic methods to conduct a cost-effectiveness analysis of histamine H2-receptor antagonists. AB - A formulary decision at a health care institution was studied by using two pharmacoeconomic methods. A pharmacoeconomic study was undertaken to assess the impact of a 1995 formulary decision to designate cimetidine as the primary histamine H2-receptor antagonist (H2RA) and to restrict the use of famotidine. Consecutive patients receiving either i.v. cimetidine or famotidine for stress ulcer prophylaxis were reviewed during a two-month period in 1997, and information on demographics, dosage and duration of H2RA therapy, admission date, laboratory test values, and adverse drug reactions was collected. Data for 62 patients (43 cimetidine recipients and 19 famotidine recipients) were evaluated. Therapy was categorized as successful or failed, and the data were then evaluated by decision analysis to evaluate the cost-effectiveness of the agents and by multiattribute utility theory (MAUT) to incorporate a humanistic evaluation of the treatments, namely, the number of doses administered and the number of times dosages were changed. The decision tree revealed that the average cost of receiving cimetidine was $82.01 and the average cost of famotidine therapy was $92.45. The MAUT analysis showed that cimetidine was the preferred agent as long as cost was valued at greater than 60% of the decision-making process and efficacy remained equal between the two agents. Two pharmacoeconomic methods lent support to a formulary decision at a health care institution. PMID- 9872688 TI - Cost-effectiveness impact of iron dextran on hemodialysis patients' use of epoetin alfa and blood. AB - The cost-effectiveness impact of iron dextran administration on the use of epoetin alfa and blood in hemodialysis patients was studied. Subjects were ambulatory hemodialysis patients who had been receiving hemodialysis for at least six months before the start of an iron dextran protocol and who had been given epoetin alfa for at least four of those six months. Clinical data were collected for six months before and six months after the protocol was implemented. Successful treatment was defined as a hematocrit of 33-36%, a transferrin saturation of >10%, a ferritin concentration of >100 ng/mL, and no blood use except for acute blood loss. A total of 33 patients completed the study. Fifty units of blood were used in the first six months and nine units in the second six months. There was significant improvement in mean hematocrit, ferritin, and transferrin saturation values after the protocol began. Average epoetin alfa doses did not change significantly. There was significant improvement in success rates for ferritin and blood use and in the overall success rate. Ten patients met all success criteria in the preprotocol period, versus 27 in the postprotocol period. Monthly cost-effectiveness analysis for the preprotocol and postprotocol periods indicated costs of $1350 and $526, per successful treatment, respectively. The incremental cost-effectiveness of iron dextran was $42 per successful treatment. Iron dextran improved iron indices and reduced the need for blood transfusions but did not reduce the average dose of epoetin alfa. The additional cost of therapy per month seemed justified by the clinical benefits. PMID- 9872689 TI - Rehospitalization rates for depot antipsychotics and pharmacoeconomic implications: comparison with risperidone. PMID- 9872690 TI - Need for therapeutic guidelines for antipsychotics under a prescription cap. PMID- 9872692 TI - Cost-effectiveness and clinical outcomes of metformin or insulin add-on therapy in adults with type 2 diabetes. PMID- 9872691 TI - Cost-effectiveness of topical 0.03% flurbiprofen in outpatient cataract surgery as measured by surgical time and vitreous loss. PMID- 9872694 TI - Pharmacoeconomic impact of a weight-based heparin protocol. PMID- 9872693 TI - Diabetes care and cost of pharmacotherapy versus medical services. PMID- 9872695 TI - Outcomes evaluation of a pharmacist discharge medication teaching service. PMID- 9872696 TI - Cost and outcomes analysis of abciximab use in a community teaching hospital. PMID- 9872697 TI - Etanercept marketed for moderate, severe rheumatoid arthritis. PMID- 9872698 TI - International pharmacists adopt statement on pharmaceutical care. PMID- 9872699 TI - Educational rotations in pharmacy-based investigational drug services. PMID- 9872700 TI - The Kaiser Permanente/USC Patient Consultation Study: change in quality of life. University of Southern California. AB - The impacts of two models of pharmacist consultation on patient function and health-related quality of life (HRQOL) relative to a control model were studied. Patients in the random-assignment study and the areawide study of the Kaiser Permanente/USC Patient Consultation Study were surveyed three times over a two year period. The patients were receiving pharmaceutical services under the Kaiser Permanente model of consultation (KP model), a state model of consultation, or a control model and were stratified according to prescription drug use. A global visual-analogue scale and Short Form-36 were used to assess HRQOL. In the areawide study, only 2 of 42 comparisons of the effects of the KP or state model on HRQOL were significant; both were associated with the state model and involved only small increases. In the random-assignment study, 3 of 21 estimated effects of the KP model on HRQOL were positive and significant; 2 of these were significantly different between the KP model and the state model. Although the KP and state models of consultation were associated with some changes in HRQOL, the overall influence was not consistent and not clinically important. PMID- 9872701 TI - The Kaiser Permanente/USC Patient Consultation Study: patient satisfaction with pharmaceutical services. University of Southern California. AB - The impacts of two models of pharmacist consultation on patient satisfaction with pharmaceutical services relative to a control model were studied. Patients in the random-assignment and areawide studies of the Kaiser Permanente/USC Patient Consultation Study were asked to rate their satisfaction with six aspects of the service they received at the pharmacy. The patients were receiving pharmaceutical services under the Kaiser Permanente (KP) model of consultation, a state-mandated model, or a control model. They were assigned to risk categories according to their pattern of prescription drug use. Differences in satisfaction among the three consultation models for all patients and for each risk group and correlations between satisfaction items were determined. The KP and state models of consultation were associated with better satisfaction than the control model on five of the six satisfaction items. In the high-risk patient groups, the KP and state models were associated with higher levels of satisfaction than the control model. In the low-risk group, the state model was associated with higher levels of satisfaction than the control model; results for low-risk patients assigned to the KP model were inconsistent between the random-assignment study and the areawide study. Overall satisfaction appeared to be a suboptimal measure of satisfaction with consultation. Overall satisfaction was more closely related to satisfaction with waiting time and with whether the pharmacy staff was helpful and caring than with the three items reflecting satisfaction with the content of the consultation. Patients were more satisfied under the state model of pharmacist consultation or the KP model of consultation, which resulted in more intensive counseling for fewer patients, than under a control model. PMID- 9872702 TI - Stability of ondansetron hydrochloride and 12 medications in plastic syringes. AB - The stability and compatibility of ondansetron hydrochloride with neostigmine methylsulfate, naloxone hydrochloride, midazolam hydrochloride, fentanyl citrate, alfentanil hydrochloride, atropine sulfate, morphine sulfate, meperidine hydrochloride, propofol, droperidol, metoclopramide monohydrochloride, and glycopyrrolate were studied. Ondansetron 1.33 or 1.0 mg/mL was combined with 0.9% sodium chloride injection and each of the 12 drugs in duplicate in plastic syringes (or glass for propofol). The syringes were stored at 21.8-23.4 or 4 degrees C in the dark, except for those containing propofol, which were stored at ambient temperature. Samples were removed at 0, 4, 8, and 24 hours for analysis by high-performance liquid chromatography and pH measurement; the propofol containing samples were removed at 0, 1, 2, and 4 hours. Syringes were visually assessed for color and clarity, and particulate content was measured with a particle counter at the end of the study period. All solutions containing ondansetron retained more than 90% of their initial ondansetron concentration. Solutions containing each of the other drugs except droperidol retained more than 90% of their initial concentration of these drugs. The solutions containing droperidol retained more than 90% of their initial droperidol concentration for up to eight hours at ambient temperature but precipitated quickly at 4 degrees C. In combinations of ondansetron 1.33 or 1.0 mg/mL and 10 of 12 drugs, all drugs were stable for 24 hours in plastic syringes at 23 and 4 degrees C; ondansetron hydrochloride 1.0 mg/mL and propofol 1.0 and 5.0 mg/mL in admixtures were stable for 4 hours, and droperidol on its own and combined with ondansetron 1.0 mg/mL was stable for no more than 8 hours at ambient temperature. PMID- 9872703 TI - Potential confusion with antiretroviral drugs. PMID- 9872705 TI - Issues in the long-term treatment of panic disorder. PMID- 9872706 TI - Combined pharmacotherapy and cognitive behavior therapy in the treatment of panic disorder. AB - Cognitive behavior therapy (CBT) has been combined with pharmacotherapy in the treatment of panic disorder in three ways: (1) to treat agoraphobic symptoms in the condition of panic with agoraphobia; (2) to reduce withdrawal effects during drug taper; and (3) to treat panic attacks. Exposure treatment and pharmacotherapy have a modest additive effect, although more patients drop out of exposure therapy combined with imipramine treatment compared with exposure therapy alone. CBT reduces symptoms of withdrawal from alprazolam and other benzodiazepines and improves the outcome of drug treatment. At present, sufficient data are not available to determine whether the effects of CBT combined with drug therapy are additive in treating panic disorder. The results of a large trial are awaited. Current CBT consists of 12 sessions and is not widely offered to patients because of cost considerations. Efforts are being made to decrease the number of sessions necessary by improving cognitive techniques. One of these models is the subject of an ongoing trial. Finally, efforts to educate and counsel patients in the clinical setting regarding the psychopathology of panic attacks may improve the outcome of pharmacotherapy. PMID- 9872707 TI - The long-term course of panic disorder and its predictors. AB - Whereas lifetime prevalence rates of panic disorder--as established in epidemiologic surveys--range between 1.6 and 3.5%, 1-month rates usually amount to much less than one half of the lifetime rates. This finding indicates that a substantial proportion of patients who had panic disorder at some stage in their life must have remitted. In contrast to these results, clinicians tend to regard panic disorder as a chronic condition because, as a rule, they see panic patients only several years after onset of the disorder. A number of small, prospective, long-term studies of such clinical populations indicate that after several years, between 17 and 70% of patients still have panic attacks, and between 36 and 82% have phobic avoidance. In the largest and longest follow-up study published to date, 45% of all patients showed an unremitting--although in a certain proportion waxing and waning--course, 24% followed a pattern of remissions and relapses, whereas 31% went back into a stable remission. The evidence of factors predicting the course of panic disorder in clinical populations suggests that long duration and agoraphobia at baseline--not the severity and frequency of panic attacks--are predictors of an unfavorable course. Additional studies are needed to determine whether personality factors, depression, and other variables are also of predictive relevance. Also, factors working during follow-up, such as positive and negative life events, coping behaviors, and treatment, should be considered in future studies. PMID- 9872709 TI - Treatment goals for panic disorder. AB - In light of the mounting evidence that panic disorder (PD) tends to run a chronic or recurrent course, treatment goals for PD--both specific and general--have been reviewed and a model has been developed to assess the achievement of these goals over the long-term treatment of this disorder. The role of patient education in setting treatment goals has received special attention. The Goal-Oriented Monitoring and Assessment of the Efficacy of Treatment model measures the efficacy of treatment in terms of which treatment goals have been achieved and to what extent. Success of long-term treatment of PD depends on clear formulation of realistic treatment goals; implementation of effective treatment modalities for achieving these goals; maintenance of all treatment gains; prevention of relapses, recurrences, and complications; ensuring decreased vulnerability for PD and its exacerbations; and continuity of care. The Goal-Oriented Monitoring and Assessment of the Efficacy of Treatment model was used to assess the treatment of 118 patients at a day clinic. Patients were followed up for 24 months. Most treatment goals were achieved during the initial intensive, integrative treatment, with treatment gains being successfully maintained during follow-up. The model identified often-neglected patient needs that required particular attention over the course of treatment and during follow-up. PMID- 9872708 TI - Panic disorder: long-term pharmacotherapy and discontinuation. AB - This article compares panic disorder (PD) medications and discusses long-term therapy. In a review of the literature, monoamine oxidase inhibitors (MAOIs), selective serotonin reuptake inhibitors (SSRIs), and benzodiazepines prove effective in treating PD. MAOIs treat comorbid depression; frequent side effects are dizziness and orthostatic hypotension. SSRIs are better tolerated than MAOIs, producing mild anticholinergic effects, but also producing gastrointestinal side effects and sexual dysfunction. Benzodiazepines are generally well tolerated when titrated gradually; moderate sedation is the most common short-term side effect. Long-term risks are physical dependence and withdrawal reactions. One hundred six PD patients were enrolled in a double-blind, 8-month, placebo-controlled trial of alprazolam and imipramine. In the 8-week short-term phase, daily dosages were titrated up to 10 mg/day of alprazolam and 250 mg/day of imipramine. The greatest number of dropouts occurred during this phase (lack of improvement and/or side effects). Alprazolam patients had a significantly more rapid onset of improvement and lower adverse events and attrition rates. In the 6-month maintenance period, patients continued short-term treatment. Patients receiving either alprazolam or imipramine developed tolerance to some side effects. At maintenance-phase completion, 62% of the alprazolam-group patients and 26% of both the imipramine- and placebo-group patients were panic free (p<0.01). Dosages were tapered to zero over 3 weeks; one third of the alprazolam patients could not discontinue. During the unblinded, 15-month follow-up, patients received open treatment selected by personal physicians on an as-needed basis. At the end of follow-up, all patients were reassessed. Patients who had completed both short-term and maintenance phases were far more likely to be panic-free (85% vs. 55%; p<0.01). PD is chronic and recurrent, and 8 months is an effective treatment period. Maintenance treatment does not lead to tolerance, even with benzodiazepines. Dose tapering must be very gradual. Completion of a long-term maintenance program strongly predicts remission. PMID- 9872710 TI - International Study of Expert Judgement on Therapeutic Use of Benzodiazepines and Other Psychotherapeutic Medications: V. Treatment strategies in panic disorder, 1992-1997. AB - The objective of this study was to assemble expert clinical experience and judgment regarding the treatment of panic disorder in a systematic, quantitative manner, particularly with respect to changes during the past 5 years. A panel of 73 internationally recognized experts in the field of pharmacotherapy of anxiety and depression was constituted by multistage peer nomination. Sixty-six experts completed a questionnaire in 1992, and 51 of those completed a follow-up questionnaire in 1997. This report focuses on the experts' responses to questions about therapeutic options as they relate to a vignette describing a typical case of panic disorder. The preferred initial treatment strategy in 1992 (59%) and in 1997 (55%) was a combination of medication with cognitive behavioral therapy. The vast majority of the expert panel included a medication in their recommendations- 91% in 1992 and 90% in 1997. Experts recommending a medication for panic in 1992 chose as first-line treatment a benzodiazepine (35%), a selective serotonin reuptake inhibitor (SSRI, 7%), an older antidepressant (33%), or a combination of medications (25%), principally a benzodiazepine plus an older antidepressant (19%). In 1997, fewer chose a benzodiazepine (15%) or an older antidepressant (11%) alone, whereas 33% chose an SSRI alone. More experts chose a combination of medications in 1997 (39%), and the increase was attributable mainly to the choice of a benzodiazepine plus an SSRI (17%). Overall, there was only a small decline in recommendations for benzodiazepines, whereas the increased choice of SSRIs came largely at the expense of the older antidepressants. As second-line medications for panic should their first-line choice fail, the experts in 1997 recommended a benzodiazepine (7%), an SSRI (15%), an older antidepressant (28%), or a combination of medications (50%), most often a benzodiazepine plus an older antidepressant (21%) or a benzodiazepine plus an SSRI (17%). (Experts were not asked to recommend second-line treatment in 1992). Thus, in case of unsatisfactory response, the experts' choices shifted from benzodiazepines and SSRIs alone toward the older antidepressants alone or combinations of an antidepressant plus a benzodiazepine. This report concluded that combined cognitive behavioral therapy plus medication was highly favored by the experts as the initial treatment strategy for panic disorder. Over the past 5 years, SSRIs displaced older antidepressants as the experts' choice for first-line pharmacotherapy of panic disorder. In case of an unsatisfactory response, the experts more often recommended an older antidepressant or a combination of an antidepressant plus a benzodiazepine. According to the experts' judgments, the benzodiazepines, especially combined with an antidepressant, remain mainstays of pharmacotherapy for panic disorder. PMID- 9872711 TI - The removal of mercury from dental-operatory wastewater by polymer treatment. AB - The mercury (Hg) content of dental-operatory wastewater has become an issue in many localities, and Hg removal is rapidly becoming a matter of concern for all dental clinics. This preliminary study tested the efficacy of polymers for the removal of Hg contaminants from the dental-unit wastewater stream. Two commercially available polymers were used to treat dental-operatory wastewater. Used separately, each polymer removed from 74.9% to 88.4% of the Hg from dental wastewater supernatant. The polymers used in combination, within the recommended pH range, removed up to 99.9% of the total Hg from dental-wastewater supernatant. The estimated optimal concentration of the two polymers is approximately 2.33 ml of each per liter of waste, and more than 90% of the Hg may be removed with 0.13 ml/l. Results indicate that a combination of the two polymers may sufficiently reduce Hg levels to allow discharge of clarified supernatants into public sewer systems. PMID- 9872713 TI - The effect of ambient carbon monoxide on low birth weight among children born in southern California between 1989 and 1993. AB - We evaluated the effect of carbon monoxide (CO) exposures during the last trimester of pregnancy on the frequency of low birth weight among neonates born 1989-1993 to women living in the Los Angeles, California, area. Using birth certificate data for that period, we assembled a retrospective cohort of infants whose mothers resided within 2 miles of 1 of 18 CO monitoring stations. Based on the gestational age and birth date of each child, we estimated last-trimester exposure by averaging the corresponding 3 months of daily CO concentrations registered at the monitoring station closest to the mother's residence (determined from the birth certificate). Where data were available (at 6 stations), we also averaged measurements taken daily for nitrogen dioxide and ozone and those taken at 6-day intervals for particulate matter [less than/equal to]10 microm (PM10) to approximate last-trimester exposures to other pollutants. Overall, the study cohort consisted of 125,573 singleton children, excluding infants born before 37 or after 44 weeks of gestation, those weighing below 1,000 or above 5,500 g at birth, those for whom fewer than 10 days of CO measurements were available during the last trimester, and those whose mothers suffered from hypertension, diabetes, or uterine bleeding during pregnancy. Within the cohort, 2,813 (2.2%) were low in birth weight (between 1,000 and 2,499 g). Exposure to higher levels of ambient CO (>5.5 ppm 3-month average) during the last trimester was associated with a significantly increased risk for low birth weight [odds ratio (OR) = 1.22; 95% confidence interval (CI), 1.03-1.44] after adjustment for potential confounders, including commuting habits in the monitoring area, sex of the child, level of prenatal care, and age, ethnicity, and education of the mother. PMID- 9872712 TI - The Yugoslavia Prospective Study of environmental lead exposure. AB - The Yugoslavia Prospective Study of environmental lead exposure has studied the associations between exposure to lead and pregnancy outcomes; childhood neuropsychological, behavioral, and physical development; and hematologic, renal, and cardiovascular function. The cohort comprises 577 children born to women recruited at midpregnancy in two towns in Kosovo, Yugoslavia; one town is the site of a lead smelter, refinery, and battery plant and the other is 25 miles away and relatively unexposed. A sample of these children has been followed at 6 month intervals through 7.5 years of age. Blood lead concentrations ranged from 1 to 70 microg/dl. Exposure to lead was not associated with adverse pregnancy outcomes. Exposure was associated with modest decrements in intelligence, small increases in blood pressure, higher risks of proteinuria, small increases in behavior problems, and perturbed hematopoiesis. Only at low level exposures (i.e., <16 microg/dl) were small associations with decreased height found. We discuss methodological problems that may hinder causal interpretation of these data, namely, use of blood lead concentration as an exposure measure, confounding, and town-specific associations. We conclude that while reported associations are small, collectively they lend support to the notion that lead is a toxicant with numerous adverse health effects. PMID- 9872714 TI - Human health risk assessment in relation to environmental pollution of two artificial freshwater lakes in The Netherlands. AB - A human health risk assessment has been performed in relation to recreational activities on two artificial freshwater lakes along the river Meuse in The Netherlands. Although the discharges of contaminants into the river Meuse have been reduced in the last decades, which is reflected in decreasing concentrations of pollutants in surface water and suspended matter, the levels in sediments are more persistent. Sediments of the two freshwater lakes appear highly polluted and may pose a health risk in relation to recreational activities. To quantify health risks for carcinogenic (e.g., polycyclic aromatic hydrocarbons) as well as noncarcinogenic compounds (e.g., heavy metals), an exposure assessment model was used. First, we used a standard model that solely uses data on sediment pollution as the input parameter, which is the standard procedure in sediment quality assessments in The Netherlands. The highest intake appeared to be associated with the consumption of contaminated fish and resulted in a health risk for Pb and Zn (hazard index exceeded 1). For the other heavy metals and for benzo(a)pyrene, the total averaged exposure levels were below levels of concern. Secondly, input data for a more location-specific calculation procedure were provided via analyses of samples from sediment, surface water, and suspended matter. When these data (concentrations in surface water) were taken into account, the risk due to consumption of contaminated fish decreased by more than two orders of magnitude and appeared to be negligible. In both exposure assessments, many assumptions were made that contribute to a major degree to the uncertainty of this risk assessment. However, this health risk evaluation is useful as a screening methodology for assessing the urgency of sediment remediation actions. PMID- 9872715 TI - Human health risk assessment: A case study involving heavy metal soil contamination after the flooding of the river Meuse during the winter of 1993 1994. AB - At the end of December 1993 and also at the end of January 1995, the river Meuse, one of the major rivers in Europe, flooded and river banks were inundated. We investigated the possible health risks of exposure to heavy metal concentrations in river bank soils resulting from the flooding of the river Meuse at the end of 1993. Soil and deposit samples and corresponding aerable and fodder crops were collected and analyzed for heavy metals. Although the soils of the floodplain of the river Meuse appeared severely polluted mainly by Cd and Zn, the heavy metal concentrations in the crops grown on these soils were within background ranges. Incidentally, the legal standard for Cd as endorsed by the Commodities Act was exceeded in wheat crops. The main exposure pathways for the general population were through the consumption of food crops grown on the river banks and through the direct ingestion of contaminated soils. For estimating potential human exposure in relation to soil pollution, we used a multiple pathway exposure model. For estimating the actual risk, we determined metal contents of vegetables grown in six experimental gardens. From this study, it can be concluded that there is a potential health risk for the river bank inhabitants as a consequence of Pb and Cd contaminations of the floodplain soils of the river Meuse, which are frequently inundated (averaged flooding frequency once every 2 years). PMID- 9872716 TI - Dietary exposure to polychlorinated biphenyls and dioxins from infancy until adulthood: A comparison between breast-feeding, toddler, and long-term exposure. AB - Food is the major source for polychlorinated biphenyl (PCB) and dioxin accumulation in the human body. Therefore, investigating food habits from early ages until reproductive age (25 years) is important in order to assess exposure risk for the next generation. The objective of this study was to assess the PCB/dioxin exposure and the relative contribution of different foods to total exposure during preschool age. Particularly, the importance of lactational PCB/dioxin exposure vs. dietary exposure until adulthood was investigated. A cohort of 207 children was studied from birth until preschool age. Based on 3 planar PCBs and 17 2,3,7,8-substituted dibenzo-para-dioxins (PCDDs) and dibenzofurans (PCDFs) measured in breast milk, a model was developed to calculate the cumulative toxic equivalent (TEQ) intake during breast-feeding (0-1 year). In 3. 5-year-old children, daily dietary intake of planar PCB-TEQ and dioxin-TEQ was measured with a validated food questionnaire. Cumulative TEQ intake from 1 to 5 years was estimated using the PCB- and dioxin-TEQ intake measured with the food questionnaire. Cumulative TEQ intake from 6 to 25 years was estimated using national food consumption and contamination data of PCB- and dioxin-TEQ intake. In toddlers, dairy products contributed 43% to PCB-TEQ and 50% to dioxin-TEQ intake. Meat and meat products contributed 14% and 19%, respectively, and processed foods 23% and 15%, respectively. Breast-feeding for 6 months contributed to the cumulative PCB/dioxin TEQ intake until 25 years of age, 12% in boys and 14% in girls. The daily TEQ intake per kilogram body weight is 50 times higher in breast-fed infants and three times higher in toddlers than in adults. Long-term dietary exposure to PCBs and dioxins in men and women is partly due to breast-feeding (12 and 14%, respectively). After weaning, dairy products, processed foods, and meat are major contributors of PCB and dioxin accumulation until reproductive age. Instead of discouraging breast-feeding, maternal transfer of PCBs and dioxins to the next generation must be avoided by enforcement of strict regulations for PCB and dioxin discharge and by reducing consumption of animal products and processed foods in all ages. PMID- 9872717 TI - Respiratory diseases and allergies in two polluted areas in East Germany. AB - This cross-sectional epidemiological study collected health data for 2,470 school children between 5 and 14 years of age (89% of eligible children) who had lived most of their lives in either one of two counties strongly impacted by industrial pollution (Bitterfeld and Hettstedt) or in a neighboring county without any sources of industrial pollution (Zerbst). The objective of the study was to examine whether regional differences--with respect to the occurrence of childhood respiratory diseases and symptoms or allergies--exist and, if such differences are found, whether they persist when we adjust for the effects of known risk factors such as medical and sociodemographic factors or factors related to the indoor environment. Controlling for medical, sociodemographic, and indoor factors, according to parental reports, children residing in Hettstedt have about a 50% increased lifetime prevalence for physician-diagnosed allergies, eczema, and bronchitis compared to children from Zerbst and about twice the number of respiratory symptoms such as wheeze, shortness of breath, and cough without cold. Sensitization to common aeroallergens according to skin prick tests [odds ratio (OR) = 1.38; 95% confidence interval (CI), 1.02-1.86] and specific IgE levels (OR = 1.75; CI, 1.31-2.33) was more common for children from Hettstedt than children from the nonpolluted county. Bitterfeld children, on the other hand, more often received a diagnosis of asthma and eczema than children residing in Zerbst and also showed slightly increased sensitization rates. In conclusion, industrial pollution related to mining and smelting operations in the county of Hettstedt were associated with a higher lifetime prevalence of respiratory disorders and an increased rate of allergic sensitization in children between the ages of 5 and 14 years. Further studies are needed to determine what role the high dust content of heavy metals plays in Hettstedt. PMID- 9872718 TI - A sensitive zonagenetic assay for rapid in vitro assessment of estrogenic potency of xenobiotics and mycotoxins. AB - Mounting evidence confirms that hepatic biosynthetic processes are essential for female sexual maturation in fish, which is directly controlled by estrogens. These oogenetic events (zonagenesis and vitellogenesis) are induced in both sexes by estrogens. In this paper, we report the induction of zona radiata (zr) proteins and vitellogenin in primary hepatocytes from Atlantic salmon (Salmo salar L.) exposed to xenoestrogens and mycotoxins. Cells were treated with doses of 1, 5, and 10 microM 4-nonylphenol (4-NP), o, p'-DDT, lindane ([gamma]-HCH), and bisphenol A (BPA), which all induced zr proteins and vitellogenin in an approximate dose-dependent manner. Hepatocytes were also treated with combinations of xenoestrogens at 1 or 2 microM, resulting in elevated levels of both zr proteins and vitellogenin, compared to single treatment. The estrogenic activity of the mycotoxin zearalenone (ZEA) and its metabolites [alpha]-ZEA) and ss-zearalenol (ss-ZEA)], with regard to zonagenesis and vitellogenesis, was assessed in this assay system. Mycotoxins were used at concentrations of 10, 100, or 1,000 nM. All induced zr proteins and vitellogenin, with [alpha]-ZEA being the strongest inducer. When cells were treated with xenoestrogens or mycotoxins in combination with an estrogen receptor inhibitor (ICI 182,780), the induction of both zr proteins and vitellogenin was inhibited in all cases. Thus, the reported estrogen effects are bonafide estrogen responses. Zona radiata proteins were more responsive than vitellogenin to both xenoestrogens and mycotoxins. The versatility and sensitivity of the hepatocyte assay demonstrates that biosynthesis of zr proteins provides a new supplementary method for estimating xenoestrogenicity and mycotoxin action. PMID- 9872719 TI - Air pollution and health effects: A study of medical visits among children in Santiago, Chile. AB - Many epidemiological studies conducted in the last several years have reported associations between exposure to airborne particulate matter, measured as PM10 (<10 microm in diameter), and daily morbidity and mortality. However, much of the evidence involves effects on the elderly population; there is less evidence about the effects of particulates on children, especially those under 2 years of age. To examine these issues, we conducted time-series analyses of 2 years of daily visits to primary health care clinics in Santiago, Chile, where counts were computed for either upper or lower respiratory symptoms and for cohorts of children 3-15 years of age and below age 2. Daily PM10 and ozone measurements and meteorological variables were available from instruments located in downtown Santiago. The multiple regression analysis indicates a statistically significant association between PM10 and medical visits for lower respiratory symptoms in children ages 3-15 and in children under age 2. PM10 is also associated with medical visits related to upper respiratory symptoms in the older cohort, while ozone is associated with visits related to both lower and upper respiratory symptoms in the older cohort. For children under age 2, a 50- microg/m3 change in PM10 (the approximate interquartile range) is associated with a 4-12% increase in lower respiratory symptoms. For children 3-15 years of age, the increase in lower respiratory symptoms ranges from 3 to 9% for a 50- microg/m3 change in PM10 and 5% per 50 ppb change in ozone. These magnitudes are similar to results from studies of children undertaken in Western industrial nations. PMID- 9872720 TI - Predictors of plasma concentrations of DDE and PCBs in a group of U.S. women. AB - We evaluated predictors of plasma concentrations of dichlorodiphenyldichloroethylene (DDE), a metabolite of dichlorodiphenyltrichloroethane (DDT), and polychlorinated biphenyls (PCBs) in a group of 240 women, controls from a breast cancer case-control study nested in the Nurses' Health Study. We considered personal attributes such as age, serum cholesterol, region of residence, adiposity, lactation, and dietary intake. DDE levels increased 0.17 ppb/year of age (p = 0.0003), and PCBs increased 0.08 ppb (p = 0.0001). DDE and PCBs increased 0.20 (p = 0.02) and 0.13 ppb (p = 0.001), respectively, per 10 mg/dl serum cholesterol. Women living in the western United States had higher levels of DDE (mean = 11.0 ppb; p = 0.003), and women in the Northeast and Midwest had higher levels of PCBs (mean = 5.6 ppb; p = 0.0002) as compared to women from other parts of the country (mean DDE = 6.3; mean PCBs = 4. 5 ppb). Levels of DDE could not be predicted from consumption of meat, fish, poultry, dairy products, vegetables, fruits, and grains. There was a positive association between fish consumption and PCB concentrations among women in the Northeast and Midwest. Using data from the cases in the nested case-control study to assess the predictive ability of the models, we confirmed that the most reliable predictors of DDE were age and serum cholesterol, and the most important predictors of PCBs were age, serum cholesterol, and residence in the Midwest or Northeast. The null results for the majority of the food variables suggest that specific dietary factors, other than fish, are not currently a substantial contributor to human exposure to DDE and PCBs. PMID- 9872722 TI - Martin rodbell obituary AB - Martin Rodbell, recipient of the Nobel Prize in Physiology or Medicine in 1994, passed away 7 December 1998 after a long illness. Rodbell had a long and distinguished career in research and senior leadership positions at the National Institutes of Health. At different times in his career, he conducted research at the National Heart Institute, the National Institute of Arthritis and Metabolic Diseases (NIAMD), and the National Institute of Environmental Health Sciences (NIEHS). He served as Scientific Director of the NIEHS from 1985 to 1989. In 1994 he shared the Nobel Prize in Physiology or Medicine with Alfred Gilman, Professor and Chairman in the Department of Pharmacology at University of Texas Southwestern Medical Center in Dallas, for their work on G-proteins, a key component of the communication system that regulates cellular activity. Rodbell was the fifth NIH intramural scientist to be awarded a Nobel Prize. PMID- 9872721 TI - Estimates of the proportion of chemicals that were carcinogenic or anticarcinogenic in bioassays conducted by the National Toxicology Program. AB - Estimates were made of the proportion of chemicals that were carcinogenic, anticarcinogenic, or either in 397 long-term bioassays conducted by the National Toxicology Program (NTP). The estimates were obtained from the global pattern of p-values obtained from statistical tests applied to individual experiments. These tests accounted for multiple comparisons using a randomization procedure and were found to operate at the correct level of significance. Representative estimates of the proportion of carcinogens [with 90% confidence intervals (CI)] compared to the NTP estimates were as follows: male mice, 0.32 (CI, 0.19-0.44), NTP = 0.29; female mice, 0. 28 (CI, 0.15-0.41), NTP = 0.34; male rats, 0.35 (CI, 0.23-0.47), NTP = 0.36; female rats, 0.34 (CI, 0.21-0.46), NTP = 0.28; all sexes and species, 0.59 (CI, 0.49-0.69), NTP = 0.51. Representative estimates of the proportion of anticarcinogens were as follows: male mice, 0. 34; female mice, 0.27; male rats, 0.40; female rats, 0.44; all sexes and species, 0.66. Thus, there was as much or more evidence in this study for anticarcinogenesis as carcinogenesis. Even though the estimators used were negatively biased, it was estimated that 85% of the chemicals were either carcinogenic or anticarcinogenic at some site in some sex species group. This suggests that most chemicals given at high enough doses will cause some sort of perturbation in tumor rates. PMID- 9872723 TI - Superfund Basic Research: A state's perspective on health effects research needs. PMID- 9872724 TI - NIEHS and CDC track human exposure to endocrine disruptors. PMID- 9872725 TI - Most unwanted. AB - Persistent organic pollutants (POPs) can travel thousands of miles, accumulate in the food chain, and persist in the environment, taking as long as centuries to degrade. POPs are known to play a role in birth defects, cancer, immune system dysfunction, and reproductive problems in wildlife. While the effects of POPs on human health are unclear, many researchers believe that long-term exposure contributes to increasing rates of birth defects, fertility problems, greater susceptibility to disease, diminished intelligence, and certain cancers. Twelve POPs have been identified by the United Nations Environment Programme as requiring urgent regulatory attention. They include the pesticides aldrin, chlordane, DDT, dieldrin, endrin, heptachlor, hexachlorobenzene, mirex, and toxaphene. Of the remaining three POPs, polychlorinated biphenyls are industrial products (used in electrical transformers), and dioxins and furans are unintentional by-products of industrial processes. PMID- 9872726 TI - Innovations. DNA detectives. AB - To understand the many potential causes and resulting consequences of DNA damage, scientists first need methods to detect it. Canadian scientists X. Chris Le and Michael Weinfeld, with help from U.S. molecular biologist Steven Leadon, developed a selective, sensitive technique for measuring DNA damage. The scientists combined a thymine glycol antibody with thymine glycol to selectively tag a specific type of DNA damage. They then added a second antibody with fluorescing properties, and used laser-induced fluorescence to identify the damaged portion of the tagged DNA. The fluorescence can be quantified, with higher levels of fluorescence indicating higher DNA damage. The technique was shown to find 1 damaged base in 1 billion normal bases. This level of sensitivity could allow the measurement of DNA damage resulting from clinical levels of radiation, and may allow scientists to establish a day-to-day baseline for DNA damage. From this baseline, it would be possible to ascertain the levels of damage that a cell can tolerate, as well as how much damaged it is capable of repairing on a daily basis. PMID- 9872727 TI - Chemical mixtures and human health AB - The development of risk assessment models based on both toxicological and mechanistic data is necessary to protect human health from adverse effects of exposure to environmental agents. Considering the potential for exposure to myriad substances during a person's lifetime, there are many uncertainties regarding the potential health effects associated with exposure to mixtures of chemicals. PMID- 9872728 TI - Bacterial plasmids: replication of extrachromosomal genetic elements encoding resistance to antimicrobial compounds. AB - Plasmids are self-replicating extrachromosomal DNA molecules found in Gram negative and Gram-positive bacteria as well as in some yeast and other fungi. Although most of them are covalently closed circular double-stranded DNA molecules, recently linear plasmids have been isolated from different bacteria. In general, plasmids are not essential for the survival of bacteria, but they may nevertheless encode a wide variety of genetic determinants, which permit their bacterial hosts to survive better in an adverse environment or to compete better with other microorganisms occupying the same ecological niche. The medical importance of plasmids that encode for antibiotic resistance, as well as specific virulence traits has been well documented and demonstrated the important role these bacterial genetic elements play in nature. Although they encode specific molecules required for initiation of their replication, plasmids rely on host encoded factors for their replication. Plasmid replication initiates in a predetermined cis-site called ori and can proceed either by a rolling circle or a theta replication mechanism. Some of the plasmid-encoded elements required for their replication, such antisense RNA molecules and DNA repeated sequences located close to ori, determine plasmid attributes like copy number and incompatibility. PMID- 9872729 TI - Protein dephosphorylation and the intracellular control of the cell number. AB - Regulating the cell number is critically important for the development and maintenance of a multi-cellular organism. The cell number can be altered by inducing cell proliferation and/or programmed cell death (apoptosis). These two processes are linked by cell cycle-regulatory pathways, and protein phosphorylation and proteolytic degradation play key roles in both. Protein dephosphorylation has been a rather neglected aspect of cell cycle control. Recent advances in this field make it imperative to provide a view of the cell cycle from a phosphatase's vantage point. Although a number of protein phosphatases may be instrumental in cell cycle and apoptosis control, the emphasis here will be on the prototypical Ser/Thr-specific protein phosphatase PP1. Experiments will be considered in their historical context. The major goal of this review will be to re-evaluate the hypothesis that PP1 - and other protein phosphatases - may function as negative growth regulators. Currently available evidence suggests that PP1 activity is required for exit from mitosis, yet may also block cell cycle progression and facilitate apoptosis. Where appropriate, results highlighting the role of the other major phosphatase, PP2A, will also be discussed. This review will conclude with some unresolved issues including the question whether PP1 might be a suitable target for anti-cancer strategies. PMID- 9872731 TI - A new CD44V3-containing isoform is involved in tumor cell growth and migration during human breast carcinoma progression. AB - CD44 isoforms belong to a family of cell adhesion molecules expressed on the cell surface of many tumor cells during human breast cancer progression. In this study we have analyzed the expression of CD44v3-containing isoforms [containing heparan sulfate addition sites for growth factor binding] in primary breast tumors, axillary nodal metastases and normal breast tissue. Using reverse transcriptase polymerase chain reaction (RT-PCR) followed by Southern blot, cloning, nucleotide sequencing and RT-in situ-PCR analyses, we have found that at least two CD44v3 containing isoforms, including one new species of CD44v2,deltav3-10 (deltav3 defined as a v3 exon lacking the first 24 base pairs) and another previously reported CD44v3,8-10 are preferentially expressed in human primary breast tumor and axillary nodal metastases but not in normal breast tissues. These finding suggest that these CD44v3-containing isoforms are closely associated with breast cancer metastasis. PMID- 9872730 TI - Aminoglycoside resistance mediated by the bifunctional enzyme 6'-N-aminoglycoside acetyltransferase-2"-O-aminoglycoside phosphotransferase. AB - The expression of the bifunctional aminoglycoside inactivating enzyme 6'-N aminoglycoside acetyltransferase-2"-O-aminoglycoside phosphotransferase is the most important mechanism of high-level aminoglycoside resistance in Staphylococcus and Enterococcus. The enzyme is unique because it presents two different aminoglycoside-modifying activities located in different regions of the molecule. The gene aac(6')-aph(2") which encodes the synthesis of the enzyme is present in Tn4100-like transposons which are inserted both in R plasmids and the chromosomes of aminoglycoside-resistant isolates. The genetic structure of aac(6')-aph(2")-containing isolates indicates that their origin is not clonal, but plasmid conjugation together with multiple insertion events are in the basis of the rapid spread of aminoglycoside resistance among Gram-positive bacteria. There is not any prevalent genetic linkage of aac(6')-aph(2") with other antibiotic-resistance determinant. However, most methicillin resistant Staphylococcus strains present also high-level aminoglycoside resistance as the consequence of constant antibiotic pressure. This situation could change in the next future with the reported reemergence of gentamicin-susceptible MRSA isolates. Recent data show that inhibitors of eukaryotic protein kinases inhibit as well the aminoglycoside phosphotransferase activity. This effect indicates a common structure for these two families of proteins and opens the possibility for a meaningful survey of inhibitors of 6'-N-aminoglycoside acetyltransferase-2"-O aminoglycoside phosphotransferase useful in clinical practice. PMID- 9872732 TI - Developing a snapshot of the ATP binding domain(s) of aminoglycoside phosphotransferases. AB - The aminoglycoside (AG) 3'-phosphotransferases [APH(3')s] are an important class of modifying enzymes which confer high-level resistance to those AGs actively modified by the enzymes. They catalyze the transfer of the terminal phosphate from ATP to the drug, thus preventing the AG s action at the 70S ribosome. These enzymes, which utilize ATP as a co-substrate, appear from amino acid alignments to be part of a much larger superfamily of kinases and ATP-binding proteins. Structure-function analyses have been initiated in our laboratory for APH(3')-II, whose gene was derived from transposon Tn5. Site-directed mutagenesis of the cloned APH(3')-II gene was used to genetically examine the residues in two highly conserved motifs proposed to participate in ATP binding. Several of these residues, in fact, were shown to affect the enzyme s affinity for ATP. We have also initiated studies using photoaffinity labelling of APH(3')-II with the ATP analogs, 8-azido-ATP and 2-azido-ATP. We have shown that 8-N3ATP and 2-N3ATP can be substituted for ATP in the APH(3')-II catalyzed phosphorylation of kanamycin; such findings indicate that the interaction of these photoaffinity analogs of ATP with APH(3')-II is biologically relevant. One of the best-characterized of the APH(3') enzymes is APH(3')-IIIa, the first of the group whose structure has been analyzed by x- ray crystallography. Several studies have demonstrated that this enzyme functions by a Theorell-Chance mechanism. Moreover, the architecture of the enzyme, crystallized in the presence of ADP has revealed residues in the ATP binding pocket which are likely to play important roles in catalysis. Once the results from biochemical analyses can be correlated with those from mutagenesis studies and x-ray crystallography, a clearer picture of the active site will be provided for an important class of AG-modifying enzymes and phosphotransferases. This picture will also allow a better understanding of these enzymes within the greater context of kinases and nucleotide-binding proteins. PMID- 9872733 TI - Aminoglycoside phosphotransferases: proteins, structure, and mechanism. AB - Aminoglycoside antibiotics constitute an important class of clinically useful drugs which are imperiled by the emergence of resistant organisms. Aminoglycoside resistance in the clinics is primarily due to the presence of modifying enzymes which N-acetylate, O-adenylate or O-phosphorylate the antibiotics. The latter family of enzymes are termed the aminoglycoside phosphotransferases or kinases and are the subject of this review. There are seven classes of aminoglycoside phosphotransferases (APH(3'), APH(2''), APH(3'off'), APH(6), APH(9), APH(4), APH(7'')) and many isozymes in each class, and although there is very little overall general sequence homology among these enzymes, certain signature residues and sequences are common. The recent determination of the three-dimensional structure of the broad spectrum aminoglycoside kinase APH(3')-IIIa complexed with the product ADP, in addition to mechanistic and mutagenic studies on this and related enzymes, has added a great deal to our understanding of this class of antibiotic resistance enzyme. In particular, the revelation of structural and mechanistic similarities between APHs and Ser/Thr and Tyr kinases has set the stage for future inhibition studies which could prove important in reversing aminoglycoside resistance. PMID- 9872734 TI - Superionic and metallic states of water and ammonia at giant planet conditions. AB - The phase diagrams of water and ammonia were determined by constant pressure ab initio molecular dynamic simulations at pressures (30 to 300 gigapascal) and temperatures (300 to 7000 kelvin) of relevance for the middle ice layers of the giant planets Neptune and Uranus. Along the planetary isentrope water and ammonia behave as fully dissociated ionic, electronically insulating fluid phases, which turn metallic at temperatures exceeding 7000 kelvin for water and 5500 kelvin for ammonia. At lower temperatures, the phase diagrams of water and ammonia exhibit a superionic solid phase between the solid and the ionic liquid. These simulations improve our understanding of the properties of the middle ice layers of Neptune and Uranus. PMID- 9872736 TI - Sum rules and interlayer conductivity of high-Tc cuprates AB - Analysis of the interlayer infrared conductivity of cuprate high-transition temperature superconductors reveals an anomalously large energy scale extending up to midinfrared frequencies that can be attributed to formation of the superconducting condensate. This unusual effect is observed in a va- riety of materials, including Tl2Ba2CuO6+x, La2-xSrxCuO4, and YBa2Cu3O6.6, which show an incoherent interlayer response in the normal state. Midinfrared range condensation was examined in the context of sum rules that can be formulated for the complex conductivity. One possible interpretation of these experiments is in terms of a kinetic energy change associated with the superconducting transition. PMID- 9872737 TI - Imaging electron wave functions of quantized energy levels in carbon nanotubes AB - Carbon nanotubes provide a unique system for studying one-dimensional quantization phenomena. Scanning tunneling microscopy was used to observe the electronic wave functions that correspond to quantized energy levels in short metallic carbon nanotubes. Discrete electron waves were apparent from periodic oscillations in the differential conductance as a function of the position along the tube axis, with a period that differed from that of the atomic lattice. Wave functions could be observed for several electron states at adjacent discrete energies. The measured wavelengths are in good agreement with the calculated Fermi wavelength for armchair nanotubes. PMID- 9872735 TI - Liquid morphologies on structured surfaces: from microchannels to microchips AB - Liquid microchannels on structured surfaces are built up using a wettability pattern consisting of hydrophilic stripes on a hydrophobic substrate. These channels undergo a shape instability at a certain amount of adsorbed volume, from a homogeneous state with a spatially constant cross section to a state with a single bulge. This instability is quite different from the classical Rayleigh Plateau instability and represents a bifurcation between two different morphologies of constant mean curvature. The bulge state can be used to construct channel networks that could be used as fluid microchips or microreactors. PMID- 9872738 TI - DOAS measurements of tropospheric bromine oxide in mid-latitudes AB - Episodes of elevated bromine oxide (BrO) concentration are known to occur at high latitudes in the Arctic boundary layer and to lead to catalytic destruction of ozone at those latitudes; these events have not been observed at lower latitudes. With the use of differential optical absorption spectroscopy (DOAS), locally high BrO concentrations were observed at mid-latitudes at the Dead Sea, Israel, during spring 1997. Mixing ratios peaked daily at around 80 parts per trillion around noon and were correlated with low boundary-layer ozone mixing ratios. PMID- 9872739 TI - Electrochemical principles for active control of liquids on submillimeter scales AB - Electrochemical methods were combined with redox-active surfactants to actively control the motions and positions of aqueous and organic liquids on millimeter and smaller scales. Surfactant species generated at one electrode and consumed at another were used to manipulate the magnitude and direction of spatial gradients in surface tension and guide droplets of organic liquids through simple fluidic networks. Solid microparticles could be transported across unconfined surfaces. Electrochemical control of the position of surface-active species within aqueous films of liquid supported on homogeneous surfaces was used to direct these films into periodic arrays of droplets with deterministic shapes and sizes. PMID- 9872740 TI - Quantum statistical corrections to dynamic nuclear magnetic resonance AB - A quantum statistical treatment of the chemical exchange between molecular eigenstates or conformations revealed previously unsuspected dynamic terms in the spin Hamiltonian operator that describes fast exchange. These terms resulted from the effect of nuclear spin on rotational and vibrational relaxation. With the traditional theory, an interpretation of new carbon-13 nuclear magnetic resonance measurements of the chemical shift of methylcyclohexane in solution showed fast exchange equilibrium constants that were inconsistent with the slow-exchange free energy difference and were spread over a range of 30 percent for the various carbon-13 positions. Modeling of the new terms indicated that they have the correct magnitude and temperature dependence to reconcile these inconsistencies. PMID- 9872741 TI - Chain length recognition: core-shell supramolecular assembly from oppositely charged block copolymers AB - Molecular recognition based on length was found to occur between oppositely charged pairs of flexible and randomly coiled block copolymers in an aqueous milieu. Matched pairs with the same block lengths of polyanions and polycations exclusively formed even in mixtures with different block lengths. These assemblies of the charged segments with matched chain lengths then formed larger core-shell-type supramolecular assemblies with an extremely narrow size distribution due to the strict phase separation between core- and shell-forming segments. PMID- 9872742 TI - Requirement for diverse, low-abundance peptides in positive selection of T cells. AB - Whether a single major histocompatibility complex (MHC)-bound peptide can drive the positive selection of large numbers of T cells has been a controversial issue. A diverse population of self peptides was shown to be essential for the in vivo development of CD4 T cells. Mice in which all but 5 percent of MHC class II molecules were bound by a single peptide had wild-type numbers of CD4 T cells. However, when the diversity within this 5 percent was lost, CD4 T cell development was impaired. Blocking the major peptide-MHC complex in thymus organ culture had no effect on T cell development, indicating that positive selection occurred on the diverse peptides present at low levels. This requirement for peptide diversity indicates that the interaction between self peptides and T cell receptors during positive selection is highly specific. PMID- 9872743 TI - Blockade of NMDA receptors and apoptotic neurodegeneration in the developing brain. AB - Programmed cell death (apoptosis) occurs during normal development of the central nervous system. However, the mechanisms that determine which neurons will succumb to apoptosis are poorly understood. Blockade of N-methyl-D-aspartate (NMDA) glutamate receptors for only a few hours during late fetal or early neonatal life triggered widespread apoptotic neurodegeneration in the developing rat brain, suggesting that the excitatory neurotransmitter glutamate, acting at NMDA receptors, controls neuronal survival. These findings may have relevance to human neurodevelopmental disorders involving prenatal (drug-abusing mothers) or postnatal (pediatric anesthesia) exposure to drugs that block NMDA receptors. PMID- 9872744 TI - Role of heteromer formation in GABAB receptor function. AB - Recently, GBR1, a seven-transmembrane domain protein with high affinity for gamma aminobutyric acid (GABA)B receptor antagonists, was identified. Here, a GBR1 related protein, GBR2, was shown to be coexpressed with GBR1 in many brain regions and to interact with it through a short domain in the carboxyl-terminal cytoplasmic tail. Heterologously expressed GBR2 mediated inhibition of adenylyl cyclase; however, inwardly rectifying potassium channels were activated by GABAB receptor agonists only upon coexpression with GBR1 and GBR2. Thus, the interaction of these receptors appears to be crucial for important physiological effects of GABA and provides a mechanism in receptor signaling pathways that involve a heterotrimeric GTP-binding protein. PMID- 9872745 TI - Rule learning by seven-month-old infants. AB - A fundamental task of language acquisition is to extract abstract algebraic rules. Three experiments show that 7-month-old infants attend longer to sentences with unfamiliar structures than to sentences with familiar structures. The design of the artificial language task used in these experiments ensured that this discrimination could not be performed by counting, by a system that is sensitive only to transitional probabilities, or by a popular class of simple neural network models. Instead, these results suggest that infants can represent, extract, and generalize abstract algebraic rules. PMID- 9872746 TI - Assembly and analysis of conical models for the HIV-1 core. AB - The genome of the human immunodeficiency virus (HIV) is packaged within an unusual conical core particle located at the center of the infectious virion. The core is composed of a complex of the NC (nucleocapsid) protein and genomic RNA, surrounded by a shell of the CA (capsid) protein. A method was developed for assembling cones in vitro using pure recombinant HIV-1 CA-NC fusion proteins and RNA templates. These synthetic cores are capped at both ends and appear similar in size and morphology to authentic viral cores. It is proposed that both viral and synthetic cores are organized on conical hexagonal lattices, which by Euler's theorem requires quantization of their cone angles. Electron microscopic analyses revealed that the cone angles of synthetic cores were indeed quantized into the five allowed angles. The viral core and most synthetic cones exhibited cone angles of approximately 19 degrees (the narrowest of the allowed angles). These observations suggest that the core of HIV is organized on the principles of a fullerene cone, in analogy to structures recently observed for elemental carbon. PMID- 9872747 TI - The transcriptional program in the response of human fibroblasts to serum. AB - The temporal program of gene expression during a model physiological response of human cells, the response of fibroblasts to serum, was explored with a complementary DNA microarray representing about 8600 different human genes. Genes could be clustered into groups on the basis of their temporal patterns of expression in this program. Many features of the transcriptional program appeared to be related to the physiology of wound repair, suggesting that fibroblasts play a larger and richer role in this complex multicellular response than had previously been appreciated. PMID- 9872748 TI - Regulated delivery of therapeutic proteins after in vivo somatic cell gene transfer. AB - Stable delivery of a therapeutic protein under pharmacologic control was achieved through in vivo somatic gene transfer. This system was based on the expression of two chimeric, human-derived proteins that were reconstituted by rapamycin into a transcription factor complex. A mixture of two adeno-associated virus vectors, one expressing the transcription factor chimeras and one containing erythropoietin (Epo) under the control of a promoter responsive to the transcription factor, was injected into skeletal muscle of immune-competent mice. Administration of rapamycin resulted in 200-fold induction of plasma Epo. Stable engraftment of this humanized system in immune-competent mice was achieved for 6 months with similar results for at least 3 months in a rhesus monkey. PMID- 9872749 TI - Processing of the notch ligand delta by the metalloprotease Kuzbanian. AB - Signaling by the Notch surface receptor controls cell fate determination in a broad spectrum of tissues. This signaling is triggered by the interaction of the Notch protein with what, so far, have been thought to be transmembrane ligands expressed on adjacent cells. Here biochemical and genetic analyses show that the ligand Delta is cleaved on the surface, releasing an extracellular fragment capable of binding to Notch and acting as an agonist of Notch activity. The ADAM disintegrin metalloprotease Kuzbanian is required for this processing event. These observations raise the possibility that Notch signaling in vivo is modulated by soluble forms of the Notch ligands. PMID- 9872750 TI - Plant paralog to viral movement protein that potentiates transport of mRNA into the phloem. AB - CmPP16 from Cucurbita maxima was cloned and the protein was shown to possess properties similar to those of viral movement proteins. CmPP16 messenger RNA (mRNA) is present in phloem tissue, whereas protein appears confined to sieve elements (SE). Microinjection and grafting studies revealed that CmPP16 moves from cell to cell, mediates the transport of sense and antisense RNA, and moves together with its mRNA into the SE of scion tissue. CmPP16 possesses the characteristics that are likely required to mediate RNA delivery into the long distance translocation stream. Thus, RNA may move within the phloem as a component of a plant information superhighway. PMID- 9872751 TI - Fate of Escherichia coli O157:H7 on fresh-cut apple tissue and its potential for transmission by fruit flies. AB - Pathogenic Escherichia coli O157:H7, as well as nonpathogenic strains ATCC 11775 and ATCC 23716, grew exponentially in wounds on Golden Delicious apple fruit. The exponential growth occurred over a longer time period on fruit inoculated with a lower concentration of the bacterium than on fruit inoculated with a higher concentration. The bacterium reached the maximum population supported in the wounds regardless of the initial inoculum concentrations. Populations of E. coli O157:H7 in various concentrations of sterilized apple juice and unsterilized cider declined over time and declined more quickly in diluted juice and cider. The decline was greater in the unsterilized cider than in juice, which may have resulted from the interaction of E. coli O157:H7 with natural populations of yeasts that increased with time. Experiments on the transmission of E. coli by fruit flies, collected from a compost pile of decaying apples and peaches, were conducted with strain F-11775, a fluorescent transformant of nonpathogenic E. coli ATCC 11775. Fruit flies were easily contaminated externally and internally with E. coli F-11775 after contact with the bacterium source. The flies transmitted this bacterium to uncontaminated apple wounds, resulting in a high incidence of contaminated wounds. Populations of the bacterium in apple wounds increased significantly during the first 48 h after transmission. Further studies under commercial conditions are necessary to confirm these findings. PMID- 9872753 TI - Identification of enzymes and quantification of metabolic fluxes in the wild type and in a recombinant aspergillus oryzae strain AB - Two alpha-amylase-producing strains of Aspergillus oryzae, a wild-type strain and a recombinant containing additional copies of the alpha-amylase gene, were characterized with respect to enzyme activities, localization of enzymes to the mitochondria or cytosol, macromolecular composition, and metabolic fluxes through the central metabolism during glucose-limited chemostat cultivations. Citrate synthase and isocitrate dehydrogenase (NAD) activities were found only in the mitochondria, glucose-6-phosphate dehydrogenase and glutamate dehydrogenase (NADP) activities were found only in the cytosol, and isocitrate dehydrogenase (NADP), glutamate oxaloacetate transaminase, malate dehydrogenase, and glutamate dehydrogenase (NAD) activities were found in both the mitochondria and the cytosol. The measured biomass components and ash could account for 95% (wt/wt) of the biomass. The protein and RNA contents increased linearly with increasing specific growth rate, but the carbohydrate and chitin contents decreased. A metabolic model consisting of 69 fluxes and 59 intracellular metabolites was used to calculate the metabolic fluxes through the central metabolism at several specific growth rates, with ammonia or nitrate as the nitrogen source. The flux through the pentose phosphate pathway increased with increasing specific growth rate. The fluxes through the pentose phosphate pathway were 15 to 26% higher for the recombinant strain than for the wild-type strain. PMID- 9872752 TI - Fate of free DNA and transformation of the oral bacterium Streptococcus gordonii DL1 by plasmid DNA in human saliva. AB - Competitive PCR was used to monitor the survival of a 520-bp DNA target sequence from a recombinant plasmid, pVACMC1, after admixture of the plasmid with freshly sampled human saliva. The fraction of the target remaining amplifiable ranged from 40 to 65% after 10 min of exposure to saliva samples from five subjects and from 6 to 25% after 60 min of exposure. pVACMC1 plasmid DNA that had been exposed to degradation by fresh saliva was capable of transforming naturally competent Streptococcus gordonii DL1 to erythromycin resistance, although transforming activity decreased rapidly, with a half-life of approximately 50 s. S. gordonii DL1 transformants were obtained in the presence of filter-sterilized saliva and a 1-microg/ml final concentration of pVACMC1 DNA. Addition of filter-sterilized saliva instead of heat-inactivated horse serum to S. gordonii DL1 cells induced competence, although with slightly lower efficiency. These findings indicate that DNA released from bacteria or food sources within the mouth has the potential to transform naturally competent oral bacteria. However, further investigations are needed to establish whether transformation of oral bacteria can occur at significant frequencies in vivo. PMID- 9872754 TI - Sequence analysis, overexpression, and antisense inhibition of a beta-xylosidase gene, xylA, from Aspergillus oryzae KBN616. AB - beta-Xylosidase secreted by the shoyu koji mold, Aspergillus oryzae, is the key enzyme responsible for browning of soy sauce. To investigate the role of beta xylosidase in the brown color formation, a major beta-xylosidase, XylA, and its encoding gene were characterized. beta-Xylosidase XylA was purified to homogeneity from culture filtrates of A. oryzae KBN616. The optimum pH and temperature of the enzyme were found to be 4.0 and 60 degrees C, respectively, and the molecular mass was estimated to be 110 kDa based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The xylA gene comprises 2,397 bp with no introns and encodes a protein consisting of 798 amino acids (86,475 Da) with 14 potential N-glycosylation sites. The deduced amino acid sequence shows high similarity to Aspergillus nidulans XlnD (70%), Aspergillus niger XlnD (64%), and Trichoderma reesei BxII (63%). The xylA gene was overexpressed under control of the strong and constitutive A. oryzae TEF1 promoter. One of the A. oryzae transformants produced approximately 13 times more of the enzyme than did the host strain. The partial-length antisense xylA gene expressed under control of the A. oryzae TEF1 promoter decreased the beta-xylosidase level in A. oryzae to about 20% of that of the host strain. PMID- 9872756 TI - Potential role of thiobacillus caldus in arsenopyrite bioleaching AB - We investigated the potential role of the three strains of Thiobacillus caldus (KU, BC13, and C-SH12) in arsenopyrite leaching in combination with a moderately thermophilic iron oxidizer, Sulfobacillus thermosulfidooxidans. Pure cultures of T. caldus and S. thermosulfidooxidans were used as well as defined mixed cultures. By measuring released iron, tetrathionate, and sulfur concentrations, we found that the presence of T. caldus KU and BC13 in the defined mixed culture lowered the concentration of sulfur, and levels of tetrathionate were comparable to or lower than those in the presence of S. thermosulfidooxidans. This suggests that T. caldus grows on the sulfur compounds that build up during leaching, increasing the arsenopyrite-leaching efficiency. This result was similar to leaching arsenopyrite with a pure culture of S. thermosulfidooxidans in the presence of yeast extract. Therefore, three possible roles of T. caldus in the leaching environment can be hypothesized: to remove the buildup of solid sulfur that can cause an inhibitory layer on the surface of the mineral, to aid heterotrophic and mixotrophic growth by the release of organic chemicals, and to solubilize solid sulfur by the production of surface-active agents. The results showed that T. caldus KU was the most efficient at leaching arsenopyrite under the conditions tested, followed by BC13, and finally C-SH12. PMID- 9872755 TI - Bacterial filament formation, a defense mechanism against flagellate grazing, is growth rate controlled in bacteria of different phyla. AB - A facultatively filamentous bacterium was isolated from eutrophic lake water and was identified as Flectobacillus sp. strain MWH38 (a member of the Cytophaga Flavobacterium-Bacteroides phylum) by comparative 16S rRNA gene sequence analysis. Filament formation by Flectobacillus sp. strain MWH38 and filament formation by Flectobacillus major, the closest known relative of strain MWH38, were studied in chemostat cultures under grazing pressure by the bacterivorous flagellate Ochromonas sp. strain DS and without predation at several growth rates. The results clearly demonstrated that filament formation by the two flectobacilli is growth rate controlled and thus independent of the presence of a predator. However, flagellate grazing positively influenced bacterial growth rates by decreasing bacterial biomass and thus indirectly stimulated filament formation. The results of investigations of cell elongation and filament formation by Comamonas acidovorans PX54 (a member of the beta subclass of the class Proteobacteria) supported the recent proposal that in this species the mechanism of filament formation is growth rate controlled. The finding that the grazing defense mechanism consisting of filament formation is growth rate controlled in the flectobacilli investigated and C. acidovorans PX54 (i.e., in bacteria belonging to divergent evolutionary phyla) may indicate that this mechanism is a phylogenetically widely distributed defense strategy against grazing. PMID- 9872757 TI - Incidence of Fusarium spp. and levels of fumonisin B1 in maize in western Kenya. AB - Maize kernel samples were collected in 1996 from smallholder farm storages in the districts of Bomet, Bungoma, Kakamega, Kericho, Kisii, Nandi, Siaya, Trans Nzoia, and Vihiga in the tropical highlands of western Kenya. Two-thirds of the samples were good-quality maize, and one-third were poor-quality maize with a high incidence of visibly diseased kernels. One hundred fifty-three maize samples were assessed for Fusarium infection by culturing kernels on a selective medium. The isolates obtained were identified to the species level based on morphology and on formation of the sexual stage in Gibberella fujikuroi mating population tests. Fusarium moniliforme (G. fujikuroi mating population A) was isolated most frequently, but F. subglutinans (G. fujikuroi mating population E), F. graminearum, F. oxysporum, F. solani, and other Fusarium species were also isolated. The high incidence of kernel infection with the fumonisin-producing species F. moniliforme indicated a potential for fumonisin contamination of Kenyan maize. However, analysis of 197 maize kernel samples by high-performance liquid chromatography found little fumonisin B1 in most of the samples. Forty seven percent of the samples contained fumonisin B1 at levels above the detection limit (100 ng/g), but only 5% were above 1,000 ng/g, a proposed level of concern for human consumption. The four most-contaminated samples, with fumonisin B1 levels ranging from 3, 600 to 11,600 ng/g, were from poor-quality maize collected in the Kisii district. Many samples with a high incidence of visibly diseased kernels contained little or no fumonisin B1, despite the presence of F. moniliforme. This result may be attributable to the inability of F. moniliforme isolates present in Kenyan maize to produce fumonisins, to the presence of other ear rot fungi, and/or to environmental conditions unfavorable for fumonisin production. PMID- 9872758 TI - Enumeration of marine viruses in culture and natural samples by flow cytometry AB - Flow cytometry (FCM) was successfully used to enumerate viruses in seawater after staining with the nucleic acid-specific dye SYBR Green-I. The technique was first optimized by using the Phaeocystis lytic virus PpV-01. Then it was used to analyze natural samples from different oceanic locations. Virus samples were fixed with 0.5% glutaraldehyde and deep frozen for delayed analysis. The samples were then diluted in Tris-EDTA buffer and analyzed in the presence of SYBR Green I. A duplicate sample was heated at 80 degreesC in the presence of detergent before analysis. Virus counts obtained by FCM were highly correlated to, although slightly higher than, those obtained by epifluorescence microscopy or by transmission electron microscopy (r = 0.937, n = 14, and r = 0.96, n = 8, respectively). Analysis of a depth profile from the Mediterranean Sea revealed that the abundance of viruses displayed the same vertical trend as that of planktonic cells. FCM permits us to distinguish between at least two and sometimes three virus populations in natural samples. Because of its speed and accuracy, FCM should prove very useful for studies of virus infection in cultures and should allow us to better understand the structure and dynamics of virus populations in natural waters. PMID- 9872760 TI - Effects of carbon substrates on nitrite accumulation in freshwater sediments AB - The contribution of the biochemical pathways nitrification, denitrification, and dissimilatory NO3- reduction to NH4+ (DNRA) to the accumulation of NO2- in freshwaters is governed by the species compositions of the bacterial populations resident in the sediments, available carbon (C) and nitrogen (N) substrates, and environmental conditions. Recent studies of major rivers in Northern Ireland have shown that high NO2- concentrations found in summer, under warm, slow-flowing conditions, arise from anaerobic NO3- reduction. Locally, agricultural pollutants entering rivers are important C and N sources, providing ideal substrates for the aquatic bacteria involved in cycling of N. In this study a range of organic C compounds commonly found in agricultural pollutants were provided as energy sources in 48-h incubation experiments to investigate if the chemical compositions of the pollutants affected which NO3- reduction pathway was followed and influenced subsequent NO2- accumulation. Carbon stored within the sediments was sufficient to support DNRA and denitrifier populations, and the resulting NO2 peak (80 &mgr;g of N liter-1 [approximate]) observed at 24 h was indicative of the simultaneous activities of both bacterial groups. The value of glycine as an energy source for denitrification or DNRA appeared to be limited, but glycine was an important source of additional N. Glucose was an efficient substrate for both the denitrification and DNRA pathways, with a NO2- peak of 160 &mgr;g of N liter 1 noted at 24 h. Addition of formate and acetate stimulated continuous NO2- production throughout the 48-h period, caused by partial inhibition of the denitrification pathway. The formate treatment resulted in a high NO2- accumulation (1,300 &mgr;g of N liter-1 [approximate]), and acetate treatment resulted in a low NO2- concentration (<100 &mgr;g of N liter-1). PMID- 9872759 TI - Use of the KlADH4 promoter for ethanol-dependent production of recombinant human serum albumin in Kluyveromyces lactis. AB - KlADH4 is a gene of Kluyveromyces lactis encoding a mitochondrial alcohol dehydrogenase activity which is specifically induced by ethanol. The promoter of this gene was used for the expression of heterologous proteins in K. lactis, a very promising organism which can be used as an alternative host to Saccharomyces cerevisiae due to its good secretory properties. In this paper we report the ethanol-driven expression in K. lactis of the bacterial beta-glucuronidase and of the human serum albumin (HSA) genes under the control of the KlADH4 promoter. In particular, we studied the extracellular production of recombinant HSA (rHSA) with integrative and replicative vectors and obtained a significant increase in the amount of the protein with multicopy vectors, showing that no limitation of KlADH4 trans-acting factors occurred in the cells. By deletion analysis of the promoter, we identified an element (UASE) which is sufficient for the induction of KlADH4 by ethanol and, when inserted in the respective promoters, allows ethanol-dependent activation of other yeast genes, such as PGK and LAC4. We also analyzed the effect of medium composition on cell growth and protein secretion. A clear improvement in the production of the recombinant protein was achieved by shifting from batch cultures (0.3 g/liter) to fed-batch cultures (1 g/liter) with ethanol as the preferred carbon source. PMID- 9872761 TI - Characterization of a facultatively psychrophilic bacterium, vibrio rumoiensis sp. nov., that exhibits high catalase activity AB - A novel facultatively psychrophilic bacterium, strain S-1, which exhibits extraordinarily high catalase activity was isolated from the drain pool of a fish product processing plant that uses H2O2 as a bleaching and microbicidal agent. The catalase activity of the isolate was 1 or 2 orders of magnitude higher than those of Corynebacterium glutamicum, Staphylococcus aureus, Pseudomonas fluorescens, and five other species tested in this study. The strain seemed to possess only one kind of catalase, according to the results of polyacrylamide gel electrophoresis of the cell extract. The optimum temperature for catalase activity was about 30 degreesC, which was about 20 degreesC lower than that for bovine catalase activity. Electron microscopic observation revealed that the surface of the microorganism was covered by blebs. Although the isolate was nonflagellated, its taxonomic position on the basis of physiological and biochemical characteristics and analysis of 16S rRNA sequence and DNA-DNA relatedness data indicated that strain S-1 is a new species belonging to the genus Vibrio. Accordingly, we propose the name Vibrio rumoiensis. The type strain is S-1 (FERM P-14531). PMID- 9872762 TI - A mechanism of resistance to hydrogen peroxide in Vibrio rumoiensis S-1. AB - A possible mechanism of resistance to hydrogen peroxide (H2O2) in Vibrio rumoiensis, isolated from the H2O2-rich drain pool of a fish processing plant, was examined. When V. rumoiensis cells were inoculated into medium containing either 5 mM or no H2O2, they grew in similar manners. A spontaneous mutant strain, S-4, derived from V. rumoiensis and lacking catalase activity did not grow at all in the presence of 5 mM H2O2. These results suggest that catalase is inevitably involved in the resistance and survival of V. rumoiensis in the presence of H2O2. Catalase activity was constitutively present in V. rumoiensis cells grown in the absence of H2O2, and its occurrence was dependent on the age of the cells, a characteristic which is observed for the HP II-type catalase of Escherichia coli. The presence of the HP II-type catalase in V. rumoiensis cells was evidenced by partial sequencing of the gene encoding the HP II-type catalase from this organism. A notable difference between V. rumoiensis and E. coli is that catalase is accumulated at very high levels ( approximately 2% of the total soluble proteins) in V. rumoiensis, in contrast to the case for E. coli. When V. rumoiensis cells which had been exposed to 5 mM H2O2 were centrifuged, most intracellular proteins, including catalase, were recovered in the medium. On the other hand, when V. rumoiensis cells were grown on plates containing various concentrations of H2O2, individual cells had a colony-forming ability inferior to those of E. coli, Bacillus subtilis, and Vibrio parahaemolyticus. Thus, it is suggested that when V. rumoiensis cells are exposed to high concentrations of H2O2, most cells will immediately be broken by H2O2. In addition, the cells which have had little or no damage will start to grow in a medium where almost all H2O2 has been decomposed by the catalase released from broken cells. PMID- 9872763 TI - In situ, real-time catabolic gene expression: extraction and characterization of naphthalene dioxygenase mRNA transcripts from groundwater. AB - We developed procedures for isolating and characterizing in situ-transcribed mRNA from groundwater microorganisms catabolizing naphthalene at a coal tar waste contaminated site. Groundwater was pumped through 0.22-microm-pore-size filters, which were then frozen in dry ice-ethanol. RNA was extracted from the frozen filters by boiling sodium dodecyl sulfate lysis and acidic phenol-chloroform extraction. Transcript characterization was performed with a series of PCR primers designed to amplify nahAc homologs. Several primer pairs were found to amplify nahAc homologs representing the entire diversity of the naphthalene degrading genes. The environmental RNA extract was reverse transcribed, and the resultant mixture of cDNAs was amplified by PCR. A digoxigenin-labeled probe mixture was produced by PCR amplification of groundwater cDNA. This probe mixture hybridized under stringent conditions with the corresponding PCR products from naphthalene-degrading bacteria carrying a variety of nahAc homologs, indicating that diverse dioxygenase transcripts had been retrieved from groundwater. Diluted and undiluted cDNA preparations were independently amplified, and 28 of the resulting PCR products were cloned and sequenced. Sequence comparisons revealed two major groups related to the dioxygenase genes ndoB and dntAc, previously cloned from Pseudomonas putida NCIB 9816-4 and Burkholderia sp. strain DNT, respectively. A distinctive subgroup of sequences was found only in experiments performed with the undiluted cDNA preparation. To our knowledge, these results are the first to directly document in situ transcription of genes encoding naphthalene catabolism at a contaminated site by indigenous microorganisms. The retrieved sequences represent greater diversity than has been detected at the study site by culture-based approaches. PMID- 9872764 TI - Highly sensitive protein translation assay for trichothecene toxicity in airborne particulates: comparison with cytotoxicity assays. AB - Screening assays for environmental mycotoxins in bulk samples currently use cytotoxicity in cell cultures, but their application to air particulate samples often lacks sensitivity and specificity for fungal spores. An assay based on inhibition of protein synthesis using translation of firefly luciferase in a rabbit reticulocyte system has been developed for the detection of trichothecene mycotoxins found in the spores of toxigenic fungi. Ethanol extracts of air particulates trapped on polycarbonate filters are ultrafiltered and applied at several dilutions to a translation reaction mixture. The activity of translated luciferase is measured directly in a luminometer, eliminating the need for radioisotopes and time-consuming sample processing. Parallel standard curves using a commercially available trichothecene provide for expression of the results in T-2 toxin equivalents per cubic meter of air. The assay can be completed in 2 h and is readily applicable to multiple samples. Comparison to the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide cytotoxicity assay indicates a 400-fold increase in sensitivity of trichothecene detection in addition to a much higher specificity for these toxins. Initial field testing indicates a strong correlation between the measured level of toxicity and the presence of toxigenic fungi detected with microbiological methods. In conclusion, this luciferase translation assay offers a rapid and highly sensitive and specific method for quantitative detection of trichothecene mycotoxin activity in air particulate samples. PMID- 9872765 TI - Effect of toxic metals on indigenous soil beta-subgroup proteobacterium ammonia oxidizer community structure and protection against toxicity by inoculated metal resistant bacteria. AB - Contamination of soils with toxic metals is a major problem on military, industrial, and mining sites worldwide. Of particular interest to the field of bioremediation is the selection of biological markers for the end point of remediation. In this microcosm study, we focus on the effect of addition of a mixture of toxic metals (cadmium, cobalt, cesium, and strontium as chlorides) to soil on the population structure and size of the ammonia oxidizers that are members of the beta subgroup of the Proteobacteria (beta-subgroup ammonia oxidizers). In a parallel experiment, the soils were also treated by the addition of five strains of metal-resistant heterotrophic bacteria. Effects on nitrogen cycling were measured by monitoring the NH3 and NH4+ levels in soil samples. The gene encoding the alpha-subunit of ammonia monooxygenase (amoA) was selected as a functional molecular marker for the beta-subgroup ammonia oxidizing bacteria. Community structure comparisons were performed with clone libraries of PCR amplified fragments of amoA recovered from contaminated and control microcosms for 8 weeks. Analysis was performed by restriction digestion and sequence comparison. The abundance of ammonia oxidizers in these microcosms was also monitored by competitive PCR. All amoA gene fragments recovered grouped with sequences derived from cultured Nitrosospira. These comprised four novel sequence clusters and a single unique clone. Specific changes in the community structure of beta-subgroup ammonia oxidizers were associated with the addition of metals. These changes were not seen in the presence of the inoculated metal-resistant bacteria. Neither treatment significantly altered the total number of beta subgroup ammonia-oxidizing cells per gram of soil compared to untreated controls. Following an initial decrease in concentration, ammonia began to accumulate in metal-treated soils toward the end of the experiment. PMID- 9872766 TI - Thermal gradient gel electrophoresis analysis of bioprotection from pollutant shocks in the activated sludge microbial community. AB - We used a culture-independent approach, namely, thermal gradient gel electrophoresis (TGGE) analysis of ribosomal sequences amplified directly from community DNA, to determine changes in the structure of the microbial community following phenol shocks in the highly complex activated sludge ecosystem. Parallel experimental model sewage plants were given shock loads of chlorinated and methylated phenols and simultaneously were inoculated (i) with a genetically engineered microorganism (GEM) able to degrade the added substituted phenols or (ii) with the nonengineered parental strain. The sludge community DNA was extracted, and 16S rDNA was amplified and analyzed by TGGE. To allow quantitative analysis of TGGE banding patterns, they were normalized to an external standard. The samples were then compared with each other for similarity by using the coefficient of Dice. The Shannon index of diversity, H, was calculated for each sludge sample, which made it possible to determine changes in community diversity. We observed a breakdown in community structure following shock loads of phenols by a decrease in the Shannon index of diversity from 1.13 to 0.22 in the noninoculated system. Inoculation with the GEM (Pseudomonas sp. strain B13 SN45RE) effectively protected the microbial community, as indicated by the maintenance of a high diversity throughout the shock load experiment (H decreased from 1.03 to only 0.82). Inoculation with the nonengineered parental strain, Pseudomonas sp. strain B13, did not protect the microbial community from being severely disturbed; H decreased from 1.22 to 0.46 for a 3-chlorophenol-4 methylphenol shock and from 1.03 to 0.70 for a 4-chlorophenol-4-methylphenol shock. The catabolic trait present in the GEM allowed for bioprotection of the activated sludge community from breakdown caused by toxic shock loading. In-depth TGGE analysis with similarity and diversity algorithms proved to be a very sensitive tool to monitor changes in the structure of the activated sludge microbial community, ranging from subtle shifts during adaptation to laboratory conditions to complete collapse following pollutant shocks. PMID- 9872767 TI - Threonine overproduction in yeast strains carrying the HOM3-R2 mutant allele under the control of different inducible promoters. AB - The HOM3 gene of Saccharomyces cerevisiae codes for aspartate kinase, which plays a crucial role in the regulation of the metabolic flux that leads to threonine biosynthesis. With the aim of obtaining yeast strains able to overproduce threonine in a controlled way, we have placed the HOM3-R2 mutant allele, which causes expression of a feedback-insensitive enzyme, under the control of four distinctive regulatable yeast promoters, namely, PGAL1, PCHA1, PCYC1-HSE2, and PGPH1. The amino acid contents of strains bearing the different constructs were analyzed both under repression and induction conditions. Although some differences in overall threonine production were found, a maximum of around 400 nmol/mg (dry weight) was observed. Other factors, such as excretion to the medium and activity of the catabolic threonine/serine deaminase, also affect threonine accumulation. Thus, improvement of threonine productivity by yeast cells would probably require manipulation of these and other factors. PMID- 9872768 TI - Plasmid-encoded anthranilate synthase (TrpEG) in Buchnera aphidicola from aphids of the family pemphigidae. AB - Buchnera aphidicola is an obligate intracellular symbiont of aphids. One of its proposed functions is the synthesis of essential amino acids, nutrients required by aphids but deficient in their diet of plant phloem sap. The genetic organization of the tryptophan pathway in Buchnera from proliferous aphids of the family Aphididae has previously been shown to reflect a capacity to overproduce this essential amino acid (C.-Y. Lai, L. Baumann, and P. Baumann, Proc. Natl. Acad. Sci. USA 91:3819-3823, 1994). This involved amplification of the genes for the first enzyme in the pathway, anthranilate synthase (TrpEG), on a low-copy number plasmid. Here we report on the finding and molecular characterization of TrpEG-encoding plasmids in Buchnera from aphids of the distantly related family Pemphigidae. Buchnera from Tetraneura caerulescens contained a 3.0-kb plasmid (pBTc2) that carried a single copy of trpEG and resembled trpEG plasmids of Buchnera from the Aphididae. The second plasmid (pBPs2), isolated from Buchnera of Pemphigus spyrothecae, contained a different replicon. It consisted of a putative origin of replication containing iterons and an open reading frame, designated repAC, which showed a high similarity to the gene encoding the replication initiation protein RepA of the RepA/C replicon from the broad-host range IncA/C group of plasmids. The plasmid population was heterogeneous with respect to the number of tandem repeats of a 1.8-kb unit carrying repAC1, trpG, and remnants of trpE. The two principal forms consisted of either five or six copies of this repeat and a single-copy region carrying repAC2, the putative origin of replication, and trpE. The unexpected finding of elements of the RepA/C replicon in previously characterized trpEG plasmids from Buchnera of the Aphididae suggests that a replacement of replicons has occurred during the evolution of these plasmids, which may point to a common ancestry for all Buchnera trpEG amplifications. PMID- 9872769 TI - A hemorrhagic factor (Apicidin) produced by toxic Fusarium isolates from soybean seeds. AB - Fifty-two isolates of Fusarium species were obtained from soybean seeds from various parts of Korea and identified as Fusarium oxysporum, F. moniliforme, F. semitectum, F. solani, F. graminearum, or F. lateritium. These isolates were grown on autoclaved wheat grains and examined for toxicity in a rat-feeding test. Nine cultures were toxic to rats. One of these, a culture of Fusarium sp. strain KCTC 16677, produced apicidin, an antiprotozoal agent that caused toxic effects in rats (including body weight loss; hemorrhage in the stomach, intestines, and bladder; and finally death) when rats were fed diets supplemented with 0.05 and 0.1% apicidin. The toxin was toxic to brine shrimp (the 50% lethal concentration was 40 microg/ml) and was weakly cytotoxic to human and mouse tumor cell lines. PMID- 9872771 TI - Ergosterol content in various fungal species and biocontaminated building materials AB - This paper reports the ergosterol content for microbial cultures of six filamentous fungi, three yeast species, and one actinomycete and the ergosterol levels in 40 samples of building materials (wood chip, gypsum board, and glass wool) contaminated by microorganisms. The samples were hydrolyzed in alkaline methanol, and sterols were silylated and analyzed by gas chromatography-mass spectrometry. The average ergosterol content varied widely among the fungal species over the range of 2.6 to 42 &mgr;g/ml of dry mass or 0.00011 to 17 pg/spore or cell. Ergosterol could not be detected in the actinomycete culture. The results for both the fungal cultures and building material samples supported the idea that the ergosterol content reflects the concentration of filamentous fungi but it underestimates the occurrence of yeast cells. The ergosterol content in building material samples ranged from 0.017 to 68 &mgr;g/g of dry mass of material. A good agreement between the ergosterol concentration and viable fungal concentrations was detected in the wood chip (r > 0.66, P 0.48, P 0.63, P < 0.0001). In conclusion, the ergosterol concentration could be a suitable marker for estimation of fungal concentrations in contaminated building materials with certain reservations, including the underestimation of yeast concentrations. PMID- 9872770 TI - Degradation of 3-chlorobenzoate under low-oxygen conditions in pure and mixed cultures of the anoxygenic photoheterotroph Rhodopseudomonas palustris DCP3 and an aerobic Alcaligenes species. AB - The presence or absence of molecular oxygen has been shown to play a crucial role in the degradability of haloaromatic compounds. In the present study, it was shown that anaerobic phototrophic 3-chlorobenzoate (3CBA) metabolism by Rhodopseudomonas palustris DCP3 is oxygen tolerant up to a concentration of 3 microM O2. Simultaneous oxidation of an additional carbon source permitted light dependent anaerobic 3CBA degradation at oxygen input levels which, in the absence of such an additional compound, would result in inhibition of light-dependent dehalogenation. Experiments under the same experimental conditions with strain DCP3 in coculture with an aerobic 3CBA-utilizing heterotroph, Alcaligenes sp. strain L6, revealed that light-dependent dehalogenation of 3CBA did not occur. Under both oxygen limitation (O2 < 0.1 microM) and low oxygen concentrations (3 microM O2), all the 3CBA was metabolized by the aerobic heterotroph. These data suggest that biodegradation of (halo)aromatics by photoheterotrophic bacteria such as R. palustris DCP3 may be restricted to anoxic photic environments. PMID- 9872772 TI - Glycerol overproduction by engineered saccharomyces cerevisiae wine yeast strains leads to substantial changes in By-product formation and to a stimulation of fermentation rate in stationary phase AB - Six commercial wine yeast strains and three nonindustrial strains (two laboratory strains and one haploid strain derived from a wine yeast strain) were engineered to produce large amounts of glycerol with a lower ethanol yield. Overexpression of the GPD1 gene, encoding a glycerol-3-phosphate dehydrogenase, resulted in a 1.5- to 2.5-fold increase in glycerol production and a slight decrease in ethanol formation under conditions simulating wine fermentation. All the strains overexpressing GPD1 produced a larger amount of succinate and acetate, with marked differences in the level of these compounds between industrial and nonindustrial engineered strains. Acetoin and 2,3-butanediol formation was enhanced with significant variation between strains and in relation to the level of glycerol produced. Wine strains overproducing glycerol at moderate levels (12 to 18 g/liter) reduced acetoin almost completely to 2,3-butanediol. A lower biomass concentration was attained by GPD1-overexpressing strains, probably due to high acetaldehyde production during the growth phase. Despite the reduction in cell numbers, complete sugar exhaustion was achieved during fermentation in a sugar-rich medium. Surprisingly, the engineered wine yeast strains exhibited a significant increase in the fermentation rate in the stationary phase, which reduced the time of fermentation. PMID- 9872773 TI - Sources of Listeria monocytogenes contamination in a cold-smoked rainbow trout processing plant detected by pulsed-field gel electrophoresis typing. AB - Sites of Listeria monocytogenes contamination in a cold-smoked rainbow trout (Oncorhynchus mykiss) processing plant were detected by sampling the production line, environment, and fish at different production stages. Two lots were monitored. The frequency of raw fish samples containing L. monocytogenes was low. During processing, the frequency of fish contaminated with L. monocytogenes clearly rose after brining, and the most contaminated sites of the processing plant were the brining and postbrining areas. A total of 303 isolates from the raw fish, product, and the environment were characterized by pulsed-field gel electrophoresis (PFGE). PFGE yielded nine pulsotypes, which formed four clusters. The predominating L. monocytogenes pulsotypes of the final product were associated with brining and slicing, whereas contaminants of raw fish were not detected in the final product. Air-mediated contamination in the plant could not be proved. In accordance with these results, an L. monocytogenes eradication program was planned. The use of hot steam, hot air, and hot water seemed to be useful in eliminating L. monocytogenes. None of the control samples taken in the 5 months after the eradication program was implemented contained L. monocytogenes. PMID- 9872775 TI - Bacterial adhesion to soil contaminants in the presence of surfactants AB - It has been proposed that addition of surfactants to contaminated soil enhances the solubility of target compounds; however, surfactants may simultaneously reduce the adhesion of bacteria to hydrophobic surfaces. If the latter mechanism is important for the biodegradation of virtually insoluble contaminants, then the use of surfactants may not be beneficial. The adhesion of a Mycobacterium strain and a Pseudomonas strain, isolated from a creosote-contaminated soil, to the surfaces of highly viscous non-aqueous-phase liquids (NAPLs) was measured. The NAPLs were organic material extracted from soils from two creosote-contaminated sites and two petroleum-contaminated sites. Cells suspended in media with and without surfactant were placed in test tubes coated with an NAPL, and the percentages of cells that adhered to the surface of the NAPL in the presence and absence of surfactant were compared by measuring optical density. Test tubes without NAPLs were used as controls. The presence of either Triton X-100 or Dowfax 8390 at a concentration that was one-half the critical micelle concentration (CMC) inhibited adhesion of both species of bacteria to the NAPLs. Both surfactants, when added at concentrations that were one-half the CMCs to test tubes containing previously adhered bacteria, also promoted the removal of the cells from the surfaces of the NAPL-coated test tubes. Neither surfactant was toxic to the bacteria. Further investigation showed that a low concentration of surfactant also inhibited the growth of both species on anthracene, indicating that the presence of a surfactant resulted in a reduction in the uptake of the solid carbon source. PMID- 9872774 TI - Morphological and biochemical properties of a Sphaerotilus sp. Isolated from paper mill slimes. AB - Four strains of filamentous bacteria were isolated from slimes collected in different paper mill factories. Morphological and physiological characterization of the isolates indicated an affiliation with the genus Sphaerotilus. However, while the physiological properties of the isolates were almost identical, pronounced physiological differences between the isolates and Sphaerotilus natans DSM 6575(T), DSM 565, and DSM 566 with respect to their ability to metabolize complex polysaccharides, sugars, polyalcohols, or organic acids as carbon sources were detected. In contrast to the analyzed culture collection strains of S. natans, all paper mill isolates were able to grow at elevated temperatures of up to 40 degrees C. Comparative sequence analysis of nearly complete 16S ribosomal DNA (rDNA) sequences from the four new isolates demonstrated that the retrieved sequences were highly similar to each other (99.6 to 99.8% similarity) and to previously published partial 16S rDNA sequences of S. natans DSM 6575(T) and ATCC 15291. Polyphasic characterization of the isolated Sphaerotilus strains revealed interesting adaptations of the strains to the environmental paper mill conditions with regard to temperature tolerance and utilization of cellulose and starch. PMID- 9872776 TI - In situ population dynamics of bacterial viruses in a terrestrial environment AB - Predation by bacteriophages is thought to control bacterial numbers and facilitate gene transfer among bacteria in the biosphere. A thorough understanding of phage population dynamics is therefore necessary if their significance in natural environments is to be fully appreciated. Here we describe the in situ population dynamics of three separate phage populations predating on separate bacterial species, living on the surface of field-grown sugar beet (Beta vulgaris var. Amethyst), as recorded over a 9-month period. The distributions of the three phage populations were different and fluctuated temporally in 1996 (peak density, approximately 10(3) PFU g-1). One of these populations, predating on the indigenous phytosphere bacterium Serratia liquefaciens CP6, consisted of six genetically distinct DNA phages that varied in relative abundance to the extent that an apparent temporal succession was observed between the two most abundant phages, PhiCP6-1 and PhiCP6-4. PMID- 9872777 TI - Production of biogenic Mn oxides by leptothrix discophora SS-1 in a chemically defined growth medium and evaluation of their Pb adsorption characteristics AB - Biogenic Mn oxides were produced by the bacterium Leptothrix discophora SS-1 (= ATCC 3182) in a chemically defined mineral salts medium, and the Pb binding and specific surface area of these oxides were characterized. Growth of SS-1 in the defined medium with pyruvate as a carbon and energy source required the addition of vitamin B12. Complete oxidation of Mn(II) within 60 h required the addition of >/=0.1 &mgr;M FeSO4. Pb adsorption isotherms were determined for the biogenic Mn oxides (and associated cells with their extracellular polymer) and compared to the Pb adsorption isotherms of cells and exopolymer alone, as well as to abiotic Mn oxides. The Pb adsorption to cells and exopolymer with biogenic Mn oxides (0.8 mmol of Mn per g) at pH 6.0 and 25 degreesC was 2 orders of magnitude greater than the Pb adsorption to cells and exopolymer alone (on a dry weight basis). The Pb adsorption to the biogenic Mn oxide was two to five times greater than the Pb adsorption to a chemically precipitated abiotic Mn oxide and several orders of magnitude greater than the Pb adsorption to two commercially available crystalline MnO2 minerals. The N2 Brunauer-Emmet-Teller specific surface areas of the biogenic Mn oxide and fresh Mn oxide precipitate (224 and 58 m2/g, respectively) were significantly greater than those of the commercial Mn oxide minerals (0.048 and 4. 7 m2/g). The Pb adsorption capacity of the biogenic Mn oxide also exceeded that of a chemically precipitated colloidal hydrous Fe oxide under similar solution conditions. These results show that amorphous biogenic Mn oxides similar to those produced by SS-1 may play a significant role in the control of trace metal phase distribution in aquatic systems. PMID- 9872778 TI - Microbial desulfurization of a crude oil middle-distillate fraction: analysis of the extent of sulfur removal and the effect of removal on remaining sulfur. AB - Rhodococcus sp. strain ECRD-1 was evaluated for its ability to desulfurize a 232 to 343 degrees C middle-distillate (diesel range) fraction of Oregon basin (OB) crude oil. OB oil was provided as the sole source of sulfur in batch cultures, and the extent of desulfurization and the chemical fate of the residual sulfur in the oil after treatment were determined. Gas chromatography (GC), flame ionization detection, and GC sulfur chemiluminesce detection analysis were used to qualitatively evaluate the effect of Rhodococcus sp. strain ECRD-1 treatment on the hydrocarbon and sulfur content of the oil, respectively. Total sulfur was determined by combustion of samples and measurement of released sulfur dioxide by infrared absorption. Up to 30% of the total sulfur in the middle distillate cut was removed, and compounds across the entire boiling range of the oil were affected. Sulfur K-edge X-ray absorption-edge spectroscopy was used to examine the chemical state of the sulfur remaining in the treated OB oil. Approximately equal amounts of thiophenic and sulfidic sulfur compounds were removed by ECRD-1 treatment, and over 50% of the sulfur remaining after treatment was in an oxidized form. The presence of partially oxidized sulfur compounds indicates that these compounds were en route to desulfurization. Overall, more than two-thirds of the sulfur had been removed or oxidized by the microbial treatment. PMID- 9872779 TI - Cloning and characterization of the polyhydroxybutyrate depolymerase gene of Pseudomonas stutzeri and analysis of the function of substrate-binding domains. AB - The extracellular polyhydroxybutyrate (PHB) depolymerase gene (phaZPst) of Pseudomonas stutzeri was cloned and sequenced. phaZPst was composed of 1,728 bp encoding a protein of 576 amino acids. Analyses of the N-terminal amino acid sequence and the matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrum of the purified enzyme showed that the mature enzyme consisted of 538 amino acids with a deduced molecular mass of 57,506 Da. Analysis of the deduced amino acid sequence of the protein revealed a domain structure containing a catalytic domain, putative linker region, and two putative substrate binding domains (SBDI and SBDII). The putative linker region was similar to the repeating units of the cadherin-like domain of chitinase A from Vibrio harveyi and chitinase B from Clostridium paraputrificum. The binding characteristics of SBDs to poly([R]-3-hydroxybutyrate) [P(3HB)] and chitin granules were characterized by using fusion proteins of SBDs with glutathione S-transferase (GST). These GST fusion proteins with SBDII and SBDI showed binding activity toward P(3HB) granules but did not bind on chitin granules. It has been suggested that the SBDs of the depolymerase interact specifically with the surface of P(3HB). In addition, a kinetic analysis for the enzymatic hydrolysis of 3 hydroxybutyrate oligomers of various sizes has suggested that the catalytic domain of the enzyme recognizes at least two monomeric units as substrates. PMID- 9872780 TI - Changes in quinone profiles of hot spring microbial mats with a thermal gradient AB - The respiratory and photosynthetic quinones of microbial mats which occurred in Japanese sulfide-containing neutral-pH hot springs at different temperatures were analyzed by spectrochromatography and mass spectrometry. All of the microbial mats that developed at high temperatures (temperatures above 68 degreesC) were so called sulfur-turf bacterial mats and produced methionaquinones (MTKs) as the major quinones. A 78 degreesC hot spring sediment had a similar quinone profile. Chloroflexus-mixed mats occurred at temperatures of 61 to 65 degreesC and contained menaquinone 10 (MK-10) as the major component together with significant amounts of either MTKs or plastoquinone 9 (PQ-9). The sunlight-exposed biomats growing at temperatures of 45 to 56 degreesC were all cyanobacterial mats, in which the photosynthetic quinones (PQ-9 and phylloquinone) predominated and MK-10 was the next most abundant component in most cases. Ubiquinones (UQs) were not found or were detected in only small amounts in the biomats growing at temperatures of 50 degreesC and above, whereas the majority of the quinones of a purple photosynthetic mat growing at 34 degreesC were UQs. A numerical analysis of the quinone profiles was performed by using the following three parameters: dissimilarity index (D), microbial divergence index (MDq), and bioenergetic divergence index (BDq). A D matrix tree analysis showed that the hot spring mats consisting of the sulfur-turf bacteria, Chloroflexus spp., cyanobacteria, and purple phototrophic bacteria formed distinct clusters. Analyses of MDq and BDq values indicated that the microbial diversity of hot spring mats decreased as the temperature of the environment increased. The changes in quinone profiles and physiological types of microbial mats in hot springs with thermal gradients are discussed from evolutionary viewpoints. PMID- 9872781 TI - In situ detection of novel bacterial endosymbionts of Acanthamoeba spp. phylogenetically related to members of the order Rickettsiales. AB - Acanthamoebae are ubiquitous soil and water bactivores which may serve as amplification vehicles for a variety of pathogenic facultative bacteria and as hosts to other, presently uncultured bacterial endosymbionts. The spectrum of uncultured endosymbionts includes gram-negative rods and gram-variable cocci, the latter recently shown to be members of the Chlamydiales. We report here the isolation from corneal scrapings of two Acanthamoeba strains that harbor gram negative rod endosymbionts that could not be cultured by standard techniques. These bacteria were phylogenetically characterized following amplification and sequencing of the near-full-length 16S rRNA gene. We used two fluorescently labelled oligonucleotide probes targeting signature regions within the retrieved sequences to detect these organisms in situ. Phylogenetic analyses demonstrated that they displayed 99.6% sequence similarity and formed an independent and well separated lineage within the Rickettsiales branch of the alpha subdivision of the Proteobacteria. Nearest relatives included members of the genus Rickettsia, with sequence similarities of approximately 85 to 86%, suggesting that these symbionts are representatives of a new genus and, perhaps, family. Distance matrix, parsimony, and maximum-likelihood tree-generating methods all consistently supported deep branching of the 16S rDNA sequences within the Rickettsiales. The oligonucleotide probes displayed at least three mismatches to all other available 16S rDNA sequences, and they both readily permitted the unambiguous detection of rod-shaped bacteria within intact acanthamoebae by confocal laser-scanning microscopy. Considering the long-standing relationship of most Rickettsiales with arthropods, the finding of a related lineage of endosymbionts in protozoan hosts was unexpected and may have implications for the preadaptation and/or recruitment of rickettsia-like bacteria to metazoan hosts. PMID- 9872782 TI - Nitrogen cycling and community structure of proteobacterial beta-subgroup ammonia oxidizing bacteria within polluted marine fish farm sediments. AB - A multidisciplinary approach was used to study the effects of pollution from a marine fish farm on nitrification rates and on the community structure of ammonia oxidizing bacteria in the underlying sediment. Organic content, ammonium concentrations, nitrification rates, and ammonia oxidizer most-probable-number counts were determined in samples of sediment collected from beneath a fish cage and on a transect at 20 and 40 m from the cage. The data suggest that nitrogen cycling was significantly disrupted directly beneath the fish cage, with inhibition of nitrification and denitrification. Although visual examination indicated some slight changes in sediment appearance at 20 m, all other measurements were similar to those obtained at 40 m, where the sediment was considered pristine. The community structures of proteobacterial beta-subgroup ammonia-oxidizing bacteria at the sampling sites were compared by PCR amplification of 16S ribosomal DNA (rDNA), using primers which target this group. PCR products were analyzed by denaturing gradient gel electrophoresis (DGGE) and with oligonucleotide hybridization probes specific for different ammonia oxidizers. A DGGE doublet observed in PCR products from the highly polluted fish cage sediment sample was present at a lower intensity in the 20-m sample but was absent from the pristine 40-m sample station. Band migration, hybridization, and sequencing demonstrated that the doublet corresponded to a marine Nitrosomonas group which was originally observed in 16S rDNA clone libraries prepared from the same sediment samples but with different PCR primers. Our data suggest that this novel Nitrosomonas subgroup was selected for within polluted fish farm sediments and that the relative abundance of this group was influenced by the extent of pollution. PMID- 9872783 TI - Production of wax esters during aerobic growth of marine bacteria on isoprenoid compounds AB - This paper describes the production of isoprenoid wax esters during the aerobic degradation of 6,10,14-trimethylpentadecan-2-one and phytol by four bacteria (Acinetobacter sp. strain PHY9, Pseudomonas nautica [IP85/617], Marinobacter sp. strain CAB [DSMZ 11874], and Marinobacter hydrocarbonoclasticus [ATCC 49840]) isolated from the marine environment. Different pathways are proposed to explain the formation of these compounds. In the case of 6,10, 14-trimethylpentadecan-2 one, these esters result from the condensation of some acidic and alcoholic metabolites produced during the biodegradation, while phytol constitutes the alcohol moiety of most of the esters produced during growth on this isoprenoid alcohol. The amount of these esters formed increased considerably in N-limited cultures, in which the ammonium concentration corresponds to conditions often found in marine sediments. This suggests that the bacterial formation of isoprenoid wax esters might be favored in such environments. Although conflicting evidence exists regarding the stability of these esters in sediments, it seems likely that, under some conditions, bacterial esterification can enhance the preservation potential of labile compounds such as phytol. PMID- 9872784 TI - Population dynamics of chesapeake bay virioplankton: total-community analysis by pulsed-field gel electrophoresis AB - Recognition of viruses as the most abundant component of aquatic microbial communities has stimulated investigations of the impact of viruses on bacterio- and phytoplankton host communities. From results of field studies to date, it is concluded that in most aquatic environments, a reduction in the number of bacteria on a daily basis is caused by viral infection. However, the modest amount of in situ virus-mediated mortality may be less significant than viral infection serving to maintain clonal diversity in the host communities directly, through gene transmission (i.e., transduction), and indirectly, by elimination of numerically dominant host species. If the latter mechanism for controlling community diversity prevails, then the overall structure of aquatic viral communities would be expected to change as well over short seasonal and spatial scales. To determine whether this occurs, pulsed-field gel electrophoresis (PFGE) was used to monitor the population dynamics of Chesapeake Bay virioplankton for an annual cycle (1 year). Virioplankton in water samples collected at six stations along a transect running the length of the bay were concentrated 100 fold by ultrafiltration. Viruses were further concentrated by ultracentrifugation, and the concentrated samples were embedded in agarose. PFGE analysis of virus DNA in the agarose plugs yielded several distinct bands, ranging from 50 to 300 kb. Principal-component and cluster analyses of the virus PFGE fingerprints indicated that changes in virioplankton community structure were correlated with time, geographical location, and extent of water column stratification. From the results of this study, it is concluded that, based on the dynamic nature of the Chesapeake Bay virioplankton community structure, the clonal diversity of bacterio- and phytoplankton host communities is an important component of the virus community. PMID- 9872785 TI - Hybridization analysis of chesapeake bay virioplankton AB - It has been hypothesized that, by specifically lysing numerically dominant host strains, the virioplankton may play a role in maintaining clonal diversity of heterotrophic bacteria and phytoplankton populations. If viruses selectively lyse only those host species that are numerically dominant, then the number of a specific virus within the virioplankton would be expected to change dramatically over time and space, in coordination with changes in abundance of the host. In this study, the abundances of specific viruses in Chesapeake Bay water samples were monitored, using nucleic acid probes and hybridization analysis. Total virioplankton in a water sample was separated by pulsed-field gel electrophoresis and hybridized with nucleic acid probes specific to either single viral strains or a group of viruses with similar genome sizes. The abundances of specific viruses were inferred from the intensity of the hybridization signal. By using this technique, a virus comprising 1/1,000 of the total virioplankton abundance (ca. 10(4) PFU/ml) could be detected. Titers of either a single virus species or a group of viruses changed over time, increasing to peak abundance and then declining to low or undetectable levels, and were geographically localized in the bay. Peak signal intensities, i.e., peak abundances of virus strains, were 10 fold greater than the low background level. Furthermore, virus species were found to be restricted to a particular depth, since probes specific to viruses from bottom water did not hybridize with virus genomes from surface water at the same geographical location. Overall, changes in abundances of specific viruses within the virioplankton were episodic, supporting the hypothesis that viral infection influences, if not controls, clonal diversity within heterotrophic bacteria and phytoplankton communities. PMID- 9872787 TI - Molecular epidemiology of Campylobacter jejuni in broiler flocks using randomly amplified polymorphic DNA-PCR and 23S rRNA-PCR and role of litter in its transmission. AB - Poultry has long been cited as a reservoir for Campylobacter spp., and litter has been implicated as a vehicle in their transmission. Chicks were raised on litter removed from a broiler house positive for Campylobacter jejuni. Litter was removed from the house on days 0, 3, and 9 after birds were removed for slaughter. Chicks were raised on these three litters under controlled conditions in flocks of 25. None of these birds yielded C. jejuni in their cecal droppings through 7 weeks. Two successive flocks from the same Campylobacter-positive broiler house were monitored for Campylobacter colonization. Campylobacter jejuni prevalence rates were determined for each flock. Randomly amplified polymorphic DNA (RAPD)-PCR and 23S rRNA-PCR typing methods were used to group isolates. A high prevalence (60%) of C. jejuni in flock 1 coincided with the presence of an RAPD profile not appearing in flock 2, which had a lower rate of prevalence (28%). A 23S rRNA-PCR typing method was used to determine if strains with different RAPD profiles and different prevalence rates contained different 23S sequences. RAPD profiles detected with higher prevalence rates contained a spacer in the 23S rRNA region 100% of the time, while RAPD profiles found with lower prevalence rates contained an intervening sequence less than 2% of the time. Data suggest varying colonizing potentials of different RAPD profiles and a source other than previously used litter as a means of transmission of C. jejuni. These molecular typing methods demonstrate their usefulness, when used together, in this epidemiologic investigation. PMID- 9872786 TI - Polycyclic aromatic hydrocarbon degradation by a new marine bacterium, Neptunomonas naphthovorans gen. nov., sp. nov. AB - Two strains of bacteria were isolated from creosote-contaminated Puget Sound sediment based on their ability to utilize naphthalene as a sole carbon and energy source. When incubated with a polycyclic aromatic hydrocarbon (PAH) compound in artificial seawater, each strain also degraded 2-methylnaphthalene and 1-methylnaphthalene; in addition, one strain, NAG-2N-113, degraded 2,6 dimethylnaphthalene and phenanthrene. Acenaphthene was not degraded when it was used as a sole carbon source but was degraded by both strains when it was incubated with a mixture of seven other PAHs. Degenerate primers and the PCR were used to isolate a portion of a naphthalene dioxygenase iron-sulfur protein (ISP) gene from each of the strains. A phylogenetic analysis of PAH dioxygenase ISP deduced amino acid sequences showed that the genes isolated in this study were distantly related to the genes encoding naphthalene dioxygenases of Pseudomonas and Burkholderia strains. Despite the differences in PAH degradation phenotype between the new strains, the dioxygenase ISP deduced amino acid fragments of these organisms were 97.6% identical. 16S ribosomal DNA-based phylogenetic analysis placed these bacteria in the gamma-3 subgroup of the Proteobacteria, most closely related to members of the genus Oceanospirillum. However, morphologic, physiologic, and genotypic differences between the new strains and the oceanospirilla justify the creation of a novel genus and species, Neptunomonas naphthovorans. The type strain of N. naphthovorans is strain NAG-2N 126. PMID- 9872788 TI - Pleiotropic effects of adaptation to a single carbon source for growth on alternative substrates AB - It is frequently assumed that populations of genetically modified microorganisms will perform their intended function and then disappear from the environment due to inherent fitness disadvantages resulting from their genetic alteration. However, modified organisms used in bioremediation can be expected to adapt evolutionarily to growth on the anthropogenic substrate that they are intended to degrade. If such adaptation results in improved competitiveness for alternative, naturally occurring substrates, then this will increase the likelihood that the modified organisms will persist in the environment. In this study, bacteria capable of degrading the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) were used to test the effects of evolutionary adaptation to one substrate on fitness during growth on an alternative substrate. Twenty lineages of bacteria were allowed to evolve under abundant resource conditions on either 2,4-D or succinate as their sole carbon source. The competitiveness of each evolved line was then measured relative to that of its ancestor for growth on both substrates. Only three derived lines showed a clear drop in fitness on the alternative substrate after demonstrable adaptation to their selective substrate, while five derived lines showed significant simultaneous increases in fitness on both their selective and alternative substrates. These data demonstrate that adaptation to an anthropogenic substrate can pleiotropically increase competitiveness for an alternative natural substrate and therefore increase the likelihood that a genetically modified organism will persist in the environment. PMID- 9872789 TI - Phylogenetic affinity of a wide, vacuolate, nitrate-accumulating Beggiatoa sp. from Monterey Canyon, California, with Thioploca spp. AB - Environmentally dominant members of the genus Beggiatoa and Thioploca spp. are united by unique morphological and physiological adaptations (S. C. McHatton, J. P. Barry, H. W. Jannasch, and D. C. Nelson, Appl. Environ. Microbiol. 62:954-958, 1996). These adaptations include the presence of very wide filaments (width, 12 to 160 microm), the presence of a central vacuole comprising roughly 80% of the cellular biovolume, and the capacity to internally concentrate nitrate at levels ranging from 150 to 500 mM. Until recently, the genera Beggiatoa and Thioploca were recognized and differentiated on the basis of morphology alone; they were distinguished by the fact that numerous Thioploca filaments are contained within a common polysaccharide sheath, while Beggiatoa filaments occur singly. Vacuolate Beggiatoa or Thioploca spp. can dominate a variety of marine sediments, seeps, and vents, and it has been proposed (H. Fossing, V. A. Gallardo, B. B. Jorgensen, M. Huttel, L. P. Nielsen, H. Schulz, D. E. Canfield, S. Forster, R. N. Glud, J. K. Gundersen, J. Kuver, N. B. Ramsing, A. Teske, B. Thamdrup, and O. Ulloa, Nature [London] 374:713-715, 1995) that members of the genus Thioploca are responsible for a significant portion of total marine denitrification. In order to investigate the phylogeny of an environmentally dominant Beggiatoa sp., we analyzed complete 16S rRNA gene sequence data obtained from a natural population found in Monterey Canyon cold seeps. Restriction fragment length polymorphism analysis of a clone library revealed a dominant clone, which gave rise to a putative Monterey Beggiatoa 16S rRNA sequence. Fluorescent in situ hybridization with a sequence-specific probe confirmed that this sequence originated from wide Beggiatoa filaments (width, 65 to 85 microm). A phylogenetic tree based on evolutionary distances indicated that the Monterey Beggiatoa sp. falls in the gamma subdivision of the class Proteobacteria and is most closely related to the genus Thioploca. This vacuolate Beggiatoa-Thioploca cluster and a more distantly related freshwater Beggiatoa species cluster form a distinct phylogenetic group. PMID- 9872790 TI - New mobilizable vectors suitable for gene replacement in gram-negative bacteria and their use in mapping of the 3' end of the Xanthomonas campestris pv. campestris gum operon. AB - We describe useful vectors to select double-crossover events directly in site directed marker exchange mutagenesis in gram-negative bacteria. These vectors contain the gusA marker gene, providing colorimetric screens to identify bacteria harboring those sequences. The applicability of these vectors was shown by mapping the 3' end of the Xanthomonas campestris gum operon, involved in biosynthesis of xanthan. PMID- 9872791 TI - Phylogenetic analysis of an anaerobic, trichlorobenzene-transforming microbial consortium. AB - A culture-independent phylogenetic survey for an anaerobic trichlorobenzene transforming microbial community was carried out. Small-subunit rRNA genes were PCR amplified from community DNA by using primers specific for Bacteria or Euryarchaeota and were subsequently cloned. Application of a new hybridization based screening approach revealed 51 bacterial clone families, one of which was closely related to dechlorinating Dehalobacter species. Several clone sequences clustered to rDNA sequences obtained from a molecular study of an anaerobic aquifer contaminated with hydrocarbons and chlorinated solvents (Dojka et al., Appl. Env. Microbiol. 64:3869-3877, 1998). PMID- 9872792 TI - Rapid detection of Ophiostoma piceae and O. quercus in stained wood by PCR. AB - A rapid, sensitive, and simple method was developed to detect the sapstain fungi Ophiostoma piceae and O. quercus in stained wood. By using microwave heating for DNA extraction and PCR with internal transcribed spacer-derived-specific primers, detection was feasible within 4 h, even with DNA obtained from a single synnema. This method can easily be extended for the detection of other wood-inhabiting fungi. PMID- 9872793 TI - Influence of osmolarity and the presence of an osmoprotectant on lactococcus lactis growth and bacteriocin production AB - Growth inhibition of Lactococcus lactis provoked by increasing osmolarity is reversed when glycine betaine (GB) or its analogs are added to a defined medium. Lacticin 481 production increased sharply with growth medium osmolarity in the absence of osmoprotectant but remained unaffected when GB was supplied in media of increasing osmolarity. PMID- 9872795 TI - Bacterial resistance to ultrasonic waves under pressure at nonlethal (manosonication) and lethal (manothermosonication) temperatures. AB - The decimal reduction times of Streptococcus faecium, Listeria monocytogenes, Salmonella enteritidis, and Aeromonas hydrophila corresponding to heat treatment at 62 degrees C were 7.1, 0.34, 0.024, and 0.0096 min, and those corresponding to manosonication treatment (40 degrees C, 200 kPa, 117 microm) were 4.0, 1.5, 0.86, and 0.90 min, respectively. The manosonication decimal reduction times of the four species investigated decreased sixfold when the amplitude was increased from 62 to 150 microm and fivefold when the relative pressure was raised from 0 to 400 kPa. In L. monocytogenes, S. enteritidis, and A. hydrophila, the lethal effect of manothermosonication was the result of the addition of the lethal effects of heat and manosonication, whereas in S. faecium it was a synergistic effect. PMID- 9872796 TI - Conditions for vigorous growth on sulfide and reactor-scale cultivation protocols for the thermophilic green sulfur bacterium chlorobium tepidum AB - We describe a reactor-scale cultivation protocol for the fastest-growing and only known thermophilic member of the family Chlorobiaceae, Chlorobium tepidum. We discovered that C. tepidum would grow with sulfide as the sole electron source at rates and with final cell yields comparable to those found with thiosulfate only if the sulfide concentration was maintained below 0.1 mM and the culture redox potential was at -300 +/- 20 mV. Such was also the requirement for growth in a photobioreactor when thiosulfate (optimum level, 12 mM) was used as the preferred electron source. For cultivation of C. tepidum on a 5- to 500-ml scale, we used the system of Balch and Wolfe (Appl. Environ. Microbiol. 32:781-791, 1976) using stopper-sealed serum tubes and bottles as an alternative to the methods commonly used for the cultivation of phototrophic anaerobes and obtained consistent results. PMID- 9872794 TI - Organic solvent tolerance of Escherichia coli is independent of OmpF levels in the membrane. AB - The organic solvent tolerance of Escherichia coli was measured under conditions in which OmpF levels were controlled by various means as follows: alteration of NaCl concentration in the medium, transformation with a stress-responsive gene (marA, robA, or soxS), or disruption of the ompF gene. It was shown that solvent tolerance of E. coli did not depend upon OmpF levels in the membrane. PMID- 9872798 TI - Biosynthetic pathway of citrinin in the filamentous fungus monascus ruber as revealed by 13C nuclear magnetic resonance AB - Carbon isotope distribution of [13C]citrinin from Monascus ruber incubated with [13C]acetate revealed that the biosynthesis of the toxin originated from a tetraketide, instead of a pentaketide as has been shown for Penicillium and Aspergillus species. The production of polyketide red pigments and citrinin by M. ruber may therefore be regulated at the level of the tetraketide branch point. PMID- 9872797 TI - Homologs of aflatoxin biosynthesis genes and sequence of aflR in Aspergillus oryzae and Aspergillus sojae. AB - The presence, but not expression, of homologs of three structural genes and a regulatory gene necessary for aflatoxin biosynthesis in Aspergillus parasiticus and A. flavus was shown for A. oryzae and A. sojae. Homologs of the regulatory gene aflR were cloned and sequenced from A. oryzae and A. sojae. PMID- 9872800 TI - Bacterial leaching of metal sulfides proceeds by two indirect mechanisms via thiosulfate or via polysulfides and sulfur AB - The acid-insoluble metal sulfides FeS2, MoS2, and WS2 are chemically attacked by iron(III) hexahydrate ions, generating thiosulfate, which is oxidized to sulfuric acid. Other metal sulfides are attacked by iron(III) ions and by protons, resulting in the formation of elemental sulfur via intermediary polysulfides. Sulfur is biooxidized to sulfuric acid. This explains leaching of metal sulfides by Thiobacillus thiooxidans. PMID- 9872799 TI - The alpha subunit of toluene dioxygenase from Pseudomonas putida F1 can accept electrons from reduced FerredoxinTOL but is catalytically inactive in the absence of the beta subunit. AB - The oxygenase component of toluene dioxygenase from Pseudomonas putida F1 is an iron-sulfur protein (ISPTOL) consisting of alpha (TodC1) and beta (TodC2) subunits. Purified TodC1 gave absorbance and electron paramagnetic resonance spectra identical to those given by purified ISPTOL. TodC1 was reduced by NADH and catalytic amounts of ReductaseTOL and FerredoxinTOL. Reduced TodC1 did not oxidize toluene, and catalysis was strictly dependent on the presence of purified TodC2. PMID- 9872801 TI - Quantification of hepatitis A virus in shellfish by competitive reverse transcription-PCR with coextraction of standard RNA. AB - To quantify hepatitis A virus (HAV) in experimentally contaminated mussels, we developed an internal standard RNA with a 7-nucleotide deletion for competitive reverse transcription (RT)-PCR. Deposited directly into the sample, this standard was used both as extraction control and as quantification tool. After coextraction and competitive RT-PCR, standard and wild-type products were detected by differential hybridization with specific probes and a DNA enzyme immunoassay. The quantifiable range with this reproducible method was 10(4) to 10(7) copies of HAV/gram or 400 to 10(6) 50% tissue culture infective doses/ml. PMID- 9872802 TI - 2-Bromoethanesulfonate, sulfate, molybdate, and ethanesulfonate inhibit anaerobic dechlorination of polychlorobiphenyls by pasteurized microorganisms AB - Dechlorination of Aroclor 1242 by pasteurized microorganisms was inhibited by 2 bromoethanesulfonate (BES), sulfate, molybdate, and ethanesulfonate. Consumption of these anions and production of sulfide from BES were detected. The inhibition could not be relieved by hydrogen. Taken together these results suggest that pattern M dechlorination is mediated by spore-forming sulfidogenic bacteria. These results also suggest that BES may inhibit anaerobic dechlorination by nonmethanogens by more than one mechanism. PMID- 9872803 TI - Expression, regulation, and mode of action of the AbiG abortive infection system of lactococcus lactis subsp. cremoris UC653 AB - The abortive infection system AbiG is encoded by the lactococcal plasmid pCI750. The abiG locus (consisting of two genes, abiGi and abiGii) was examined by Northern blot analysis, revealing two transcripts of approximately 2.8 and 1.5 kb which were homologous to the two gene-specific probes. A transcriptional start site was mapped upstream of abiGi, and it appeared that the two genes were cotranscribed, resulting in the 2.8-kb transcript. The smaller transcript may be the result of independent transcription of abiGii within abiGi or of the presence of a weak terminator within abiGii. The locus was shown to be constitutively expressed. Evidence is presented for the possible existence of a second Abi mechanism on pCI750. Examination of phage sk1 RNA synthesis demonstrated that both the subcloned AbiG and, to a greater extent, pCI750 inhibited this process. pCI750 also severely inhibited synthesis of both early and late phage c2 transcripts, while the presence of the subclone resulted in a reduction in late transcript synthesis only. PMID- 9872804 TI - Sources of Vibrio mimicus contamination of turtle eggs. AB - Vibrio mimicus contamination of sand increased significantly during the arrival of the olive ridley sea turtles (Lepidochelys olivacea) at Ostional anidation beach, Costa Rica. Statistical analysis supports that eggs are contaminated with V. mimicus by contact with the sand nest. V. mimicus was isolated from eggs of all nests tested, and ctxA+ strains were found in 31% of the nests, all of which were near the estuary. PMID- 9872805 TI - Symbiotic deficiencies associated with a coxWXYZ mutant of bradyrhizobium japonicum AB - The terminal oxidase complexes encoded by coxMNOP and coxWXYZ were studied by analysis of mutations in each of the two oxidases. Carbon monoxide difference spectra obtained from membranes of coxMNOP mutant bacteroids were like those obtained for the wild type, whereas bacteroid membranes of a coxWXYZ mutant were deficient in CO-reactive cytochrome b. Experiments involving cyanide inhibition of oxidase activity were consistent with the conclusion that the coxX mutant is deficient in a membrane-associated O2-binding component. The viable cell number (bacteria that could be recultured from crushed nodules) was 20 to 29% lower for the coxX mutant than for the wild-type or the CoxN- strain. In three separate greenhouse studies, nodules of a coxX mutant had significantly lower (28 to 34% less) acetylene reduction rates than the wild-type nodules did, and plants inoculated with a double mutant (coxMNOP coxWZYZ) had rates 30% lower than those of wild-type-inoculated plants. PMID- 9872806 TI - Growth of Listeria monocytogenes and Yersinia enterocolitica on cooked modified atmosphere-packaged poultry in the presence and absence of a naturally occurring microbiota. AB - Cooked poultry cuts were inoculated with five-strain composite mixtures of either Listeria monocytogenes or Yersinia enterocolitica (1,000 CFU/150-g piece), packaged in 44:56 CO2-N2, and stored at 3.5, 6.5, or 10 degrees C for up to 5 weeks. Both L. monocytogenes and Y. enterocolitica grew under all test conditions. The presence of a naturally occurring microbiota did not influence the growth of either pathogen. Addition of lactate with the shelf life extender ALTA 2341 lengthened the lag phases of L. monocytogenes and Y. enterocolitica but did not prevent their growth. PMID- 9872807 TI - Development of a PCR assay for detection of Yersinia ruckeri in tissues of inoculated and naturally infected trout. AB - A PCR-based method was developed for the specific detection of Yersinia ruckeri in tissues of inoculated trout and naturally infected trout. No amplification products were obtained with other yersiniae, bacterial fish pathogens, or phylogenetically related bacteria (n = 34). The sensitivity of PCR detection was 60 to 65 bacterial cells per PCR tube, which was decreased to 10 to 20 cells by hybridization with a nonradioactive probe. The PCR assay proved to be as reliable as and faster than the conventional culture method for the detection of Y. ruckeri in infected trout tissues. PMID- 9872808 TI - Persistence of colonization of human colonic mucosa by a probiotic strain, Lactobacillus rhamnosus GG, after oral consumption. AB - Lactobacillus rhamnosus GG is one of the most thoroughly studied probiotic strains. Its advantages in the treatment of gastrointestinal disorders are well documented. The aim of the present study was to demonstrate with colonic biopsies the attachment of strain GG to human intestinal mucosae and the persistence of the attachment after discontinuation of GG administration. A whey drink fermented with strain GG was fed to human volunteers for 12 days. Fecal samples were collected before, during, and after consumption. L. rhamnosus GG-like colonies were detected in both fecal and colonic biopsy samples. Strain GG was identified by its characteristic colony morphology, a lactose fermentation test, and PCR. This study showed that strain GG was able to attach in vivo to colonic mucosae and, although the attachment was temporary, to remain for more than a week after discontinuation of GG administration. The results demonstrate that the study of fecal samples alone is not sufficient in evaluating colonization by a probiotic strain. PMID- 9872809 TI - Chlorine inactivation of Sphingomonas cells attached to goethite particles in drinking water. AB - Bacteria in drinking water, attached or not attached to goethite particles, were disinfected with chlorine. No additional protection was provided to the bacteria by their attachment to particles, and the limited efficiency of inactivation by chlorine was attributed to the presence of bacterial aggregates in both types of suspension. PMID- 9872811 TI - Venous thromboembolism. PMID- 9872812 TI - Interpretation of repeated tuberculin tests. Boosting, conversion, and reversion. PMID- 9872813 TI - Effect of inhaled budesonide on lung function and airway inflammation. Assessment by various inflammatory markers in mild asthma. AB - In a double-blind, cross-over study, we examined the effect of inhaled budesonide (800 microgram twice daily via Turbohaler) on lung function and various markers of airway inflammation including airway responsiveness to methacholine (PC20), exhaled nitric oxide (NO), eosinophils in induced sputum, bronchoalveolar lavage (BAL), and airway biopsies from 14 patients with mild asthma needing beta2- agonist therapy only. After inhaled steroids, there was a significant increase in FEV1 and PC20, and reduction in exhaled NO. Eosinophils in induced sputum and airway biopsy sections were also significantly decreased, although BAL eosinophil counts remained unchanged. At baseline, significant correlations were observed between exhaled NO and PC20 methacholine (r = 0.64, p < 0.05), exhaled NO and peak expiratory flow rate (PEFR) variability (r = 0. 65, p < 0.05), sputum eosinophils and FEV1 (r = -0.63, p = 0.05), and sputum eosinophils and log PC20 methacholine (r = -0.67, p < 0. 05). After treatment with inhaled steroids, there was a significant correlation between eosinophils in biopsy sections, and BAL, with log PC20 methacholine. It is likely that these parameters represent different aspects of the inflammatory process, which are all inhibited by inhaled steroids. PMID- 9872814 TI - Protective effects of inhaled PGE2 on allergen-induced airway responses and airway inflammation. AB - Inhalation of prostaglandin E2 (PGE2) had been reported to prevent allergen induced bronchoconstrictor responses; however, the effects of inhaled PGE2 on allergen-induced airway inflammation or hyperresponsiveness after allergen are unknown. This study examined the effects of inhaled PGE2 on allergen-induced airway responses and inflammation. Eight mild asthmatics with a dual airway response to inhaled allergen were recruited into a double-blind randomized crossover study comparing the effects of inhaled PGE2 (100 microgram) or placebo, on allergen-induced changes in FEV1 measured for 7 h, induced sputum inflammatory cells, obtained at baseline, 7 and 24 h, and methacholine airway responsiveness measured at 24 h after challenge. Inhaled PGE2 attenuated the allergen-induced early fall in FEV1 from 24.4 +/- 3.6% after placebo to 10.3 +/- 2.5% after PGE2 (p = 0.002), the late fall in FEV1 from 21.2 +/- 2.7% after placebo to 12.6 +/- 3.6% after PGE2 (p = 0.03), allergen-induced methacholine airway hyperresponsiveness (p = 0.03) and allergen-induced increases in percent sputum eosinophils from 36.3 +/- 8.8% after placebo to 21.0 +/- 7.3% after PGE2 (p = 0.01), percentage of EG2+ cells (p = 0.02), and percentage of metachromatic cells (p = 0.02). These results indicate that inhaled PGE2 attenuates allergen-induced airway responses, hyperresponsiveness, and inflammation, when given immediately before inhaled allergen. PMID- 9872815 TI - Intrapleural urokinase versus normal saline in the treatment of complicated parapneumonic effusions and empyema. A randomized, double-blind study. AB - Intrapleural administration of fibrinolytic agents has been shown to be effective and safe in the treatment of loculated parapneumonic pleural effusions. However, controlled studies of the possible role of the activity of urokinase (UK) through the volume effect are lacking. We therefore investigated the hypothesis that UK is effective through the lysis of pleural adhesions and not through the volume effect. Thirty-one consecutive patients with multiloculated pleural effusions were randomly assigned to receive either intrapleural UK (15 patients) or normal saline (NS) (16 patients) for 3 d, in a double-blind manner. All patients had inadequate drainage through a chest tube (< 70 ml/24 h). UK was given daily through the chest tube in a dose of 100.000 IU diluted in 100 ml of NS. Controls were given the same volume of NS intrapleurally. Response was assessed by clinical outcome, fluid drainage, chest radiography, pleural ultrasonography (US) and/or computed tomography (CT). Clinical and radiographic improvement was noted in all but two patients in the UK group but in only four in the control group. The net mean volume drained during the 3-d treatment period was significantly greater in the UK group (970 +/- 75 ml versus 280 +/- 55 ml, p < 0.001). Pleural fluid drainage was complete in 13 (86.5%) patients in the UK group (two patients were treated through video-assisted thoracoscopy) but in only four (25%) in the control group. Twelve patients in the control group were subsequently treated with UK and six of them had complete drainage; the remaining six patients had complete drainage after video-assisted thoracoscopy. Our results suggest that UK is effective in the treatment of loculated pleural effusions through the lysis of pleural adhesions and not through the volume effect. PMID- 9872816 TI - A comparison of apnea-hypopnea indices derived from different definitions of hypopnea. AB - We examined the effects of arousal- and desaturation-based scoring criteria on the apnea-hypopnea index (AHI) and on the measured prevalence of obstructive sleep apnea (OSA). Ninety-four randomly selected patients underwent overnight polysomnography. Studies were scored according to three different criteria for hypopnea, as defined by a >= 10 s discernible reduction in thoracoabdominal movement associated with: (1) >= 4% decrease in oxygen saturation (SaO2) (Type A); (2) either a >= 4% decrease in SaO2 or an arousal (Type B); or (3) electroencephalographically based arousal alone (Type C). Excellent correlation existed between AHI-A, AHI-B, and the oxygen desaturation index (ODI) (r > 0.98). AHI-A and AHI-B differed by only 2.04 +/- 1.72/h (2 SD). AHI-A and AHI-B differed from the ODI by 1.04 +/- 4.07/h and 3.07 +/- 4.30/h, respectively. Despite these small differences, use of the Type B rather than Type A definition resulted in an extra case of OSA being diagnosed for every 14 to 31 patients tested, depending on the definition of OSA (AHI: >= 5, 10, 15, or 20/h). The addition of arousal based scoring criteria for hypopnea causes only small changes in the AHI, but if OSA is defined solely by an AHI value, the measured prevalence of OSA will increase. PMID- 9872817 TI - Expiratory phase tracheal gas insufflation and pressure control in sheep with permissive hypercapnia. AB - Tracheal gas insufflation (TGI) has been shown to be a useful adjunct to mechanical ventilation, decreasing PaCO2 during permissive hypercapnia. While TGI can be used either with pressure (PCV) or volume-controlled ventilation and continuously or only during the expiratory phase (Ex-TGI), there are no controlled studies evaluating the effects of Ex-TGI with PCV in acute lung injury when the direction of the insufflated flow or the inspiratory:expiratory (I:E) ratio are varied. We evaluated the effect that Ex-TGI with PCV would have on CO2 removal during both direct and reverse insufflated flow direction with varied I:E ratios when peak airway pressure, total positive end-expiratory pressure (PEEP), and tidal volume (VT) were kept constant. In addition we examined the effect that insufflation flow directed toward the mouth (reverse flow) would have on the generation of PEEP compared with flow directed toward the carina (direct flow). After saline lavage, nine sheep were ventilated with PCV to a baseline PaCO2 of 80 mm Hg. Ex-TGI (10 L/min) was then randomly applied in the reverse and direct direction with I:E set at 1:2 or 2:1. During 1:2 I:E PaCO2 decreased from 78 +/- 4 mm Hg to 60 +/- 7 mm Hg (23.5 +/- 8.9%) with direct flow and to 64 +/- 5 mm Hg (18.5 +/- 5.5%) with reverse flow (p < 0.05), whereas during 2:1 I:E PaCO2 decreased from 80 +/- 4 mm Hg to 69 +/- 8 mm Hg (13.7 +/- 9.2%) with direct flow and to 66 +/- 4 mm Hg (17.2 +/- 4.4%) with reverse flow (p < 0.05). Greater PEEP was developed with direct flow (2.8 cm H2O I:E 1:2 and 4.0 cm H2O I:E 2:1) than with reverse flow (-0.9 cm H2O I:E 1:2 and -0.4 cm H2O I:E 2:1), p < 0.05. There was no difference in the PaCO2 change between I:E with reverse flow, but the PaCO2 decrease was greater (p < 0.05) during 1:2 versus 2:1 I:E with direct flow. CO2 removal during PCV and Ex-TGI is more consistent with reverse flow than with direct flow and PEEP level is less affected by TGI with reverse flow than with direct flow. PMID- 9872818 TI - Derivation of tumor-specific cytolytic T-cell clones from two lung cancer patients with long survival. AB - We derived lung carcinoma cell lines from tumor material resected from a patient with small-cell lung cancer (SCLC) and from a patient with non-small-cell lung cancer (NSCLC). The patient with NSCLC was vaccinated with irradiated autologous tumor cells. The two patients enjoyed an exceptionally favorable clinical evolution and are currently without signs of cancer 10 and 8 yr after their diagnoses, respectively. Autologous mixed lymphocyte-tumor cell cultures (MLTC) were produced with blood lymphocytes stimulated with irradiated autologous tumor cells. The first patient's SCLC cells, which carried a small amount of human leukocyte antigen (HLA) class I molecules, were incubated with interferon-gamma (IFN-gamma) before being used as stimulator cells. A cytolytic T-lymphocyte (CTL) clone was derived that specifically lysed the IFN-gamma-treated SCLC cells but did not lyse untreated tumor cells or autologous lymphoblasts. Clones of autologous tumor-specific CTL, directed against the NSCLC cells of the other patient, were also obtained. These tumor cells carried a higher level of HLA class I molecules and were lysed by the CTL without incubation with IFN-gamma. Altogether, these results indicate that SCLC and NSCLC cancer cells can be recognized by autologous CTL, and might therefore be susceptible to specific immunotherapy. PMID- 9872819 TI - Reconciling in vitro and in vivo measurements of aerosol delivery from a metered dose inhaler during mechanical ventilation and defining efficiency-enhancing factors. AB - We attempted to resolve the discrepancies in reported data on aerosol deposition from a chlorofluorocarbon (CFC)-propelled metered-dose inhaler (MDI) during mechanical ventilation, obtained by in vivo and in vitro methodologies. Albuterol delivery to the lower respiratory tract was decreased in a humidified versus a dry circuit (16.2 versus 30.4%, respectively; p < 0.01). In 10 mechanically ventilated patients, 4.8% of the nominal dose was exhaled. When the exhaled aerosol was subtracted from the in vitro delivery of 16.2% achieved in a humidified ventilator circuit, the resulting value (16.2 - 4.8 = 11.4%) was similar to in vivo estimates of aerosol deposition. Having reconciled in vitro with in vivo findings, we then evaluated factors influencing aerosol delivery. A lower inspiratory flow rate (40 versus 80 L/min; p < 0.001), a longer duty cycle (0.50 versus 0.25; p < 0.04), and a shorter interval between successive MDI actuations (15 versus 60 s; p < 0.02) increased aerosol delivery, whereas use of a hydrofluoroalkane (HFA)-propelled MDI decreased aerosol delivery compared with the CFC-propelled MDI. A MDI and actuator combination other than that designed by the manufacturer altered aerosol particle size and decreased drug delivery. In conclusion, aerosol delivery in an in vitro model accurately reflects in vivo delivery, providing a means for investigating methods to improve the efficiency of aerosol therapy during mechanical ventilation. PMID- 9872820 TI - A community study of exhaled nitric oxide in healthy children. AB - Exhaled nitric oxide (eNO) is elevated in patients with inflammatory pulmonary diseases and it has attracted increasing interest as a simple, noninvasive marker of airway inflammation. Little is known, however, about factors that might affect eNO in healthy subjects. We measured eNO in 157 healthy 7- to 13-yr-old children (mean 9.7 yr, 77 girls), with no history of respiratory tract disease, using a recently validated, single-breath technique. Measurements of eNO were obtained at driving (mouth) pressures of 10, 15, and 20 cm H2O and 3 eNO plateaux were achieved for each child at each pressure. Exhaled NO decreased with increasing pressure (increasing expiratory flow) (p < 0.001) and increased with age (p < 0.001). Concentrations were greater in children with a positive skin prick test (p < 0.0001). Geometric mean eNO levels were 7.2 ppb in children with no positive skin prick tests (n = 116), 10.9 ppb in children with one positive reaction (n = 24), and 20.1 ppb in children with two or more skin reactions (n = 17). Age and immunological reactions to common allergens are associated with increased eNO in children and should be controlled for in studies of eNO. The mechanisms responsible for these associations require further study. PMID- 9872821 TI - Measurements of exhaled nitric oxide with the single-breath technique and positive expiratory pressure in infants. AB - The aim of this study was to adapt the single-breath technique with positive expiratory pressure to measure exhaled nitric oxide (eNO) in infants. We hypothesized that exhaled eNO was greater in wheezy than in healthy infants. We studied 30 infants (16 wheezy and 14 healthy). The forced expiratory volume in 0.5 s (FEV0.5) was determined with the raised volume rapid thoracic compression technique, and eNO was measured during constant expiratory flow with a rapid response chemiluminescence analyzer. After passive inflation to a preset pressure of 20 cm H2O, thoracic compression with an inflatable jacket caused forced expiration to occur through a face-mask with an expiratory flow resistor attached. During the forced expiration, the jacket pressure was increased to maintain a constant driving mouth pressure and hence a constant expiratory flow (50 ml/s). The mean level of eNO in the wheezy infants (31.8 ppb) was significantly higher than the level in healthy infants (18.8 ppb) (p = 0.03). A family history of atopy in parents was associated with increased eNO levels (p < 0.001) independent of age, sex, weight, length, wheezing, and FEV0.5. We conclude that the single-breath technique with positive expiratory pressure is a feasible method for measuring eNO in infants. Levels of eNO were significantly higher in wheezy infants and in those with a family history of atopy. PMID- 9872822 TI - The effects of regular inhaled formoterol, budesonide, and placebo on mucosal inflammation and clinical indices in mild asthma. AB - The present study was designed to observe the effects of 8 wk of treatment with formoterol (Foradil) 24 microgram, budesonide 400 microgram, and matched placebo inhaled twice a day on inflammatory indices in the bronchial mucosa of 64 patients with mild atopic asthma. Biopsies were obtained at the start and 1 wk before stopping a 9-wk period of treatment, and inflammatory cell numbers were assessed in the submucosa and epithelium by immunohistochemistry. Regular formoterol significantly reduced the number of submucosal mast cells, with a similar trend for eosinophils but not activated T cells. A subgroup analysis conducted in biopsies with >= 10 eosinophils per mm2 revealed a significant reduction in eosinophil numbers when compared with both pretreatment baseline (p < 0.01) and changes after placebo (p < 0.01). Parallel, but less pronounced, effects were observed on mast cell but not on CD25(+) T cell numbers. There was no effect of any of the three treatments on BAL levels of mast cell or eosinophil mediators. We conclude that regular treatment with inhaled formoterol reduces rather than increases inflammatory cells in the mucosa of asthmatic patients. It is possible that these cellular effects of formoterol may contribute to the therapeutic efficacy of this drug when used regularly in the treatment PMID- 9872823 TI - Change in airway responsiveness among apprentices exposed to metalworking fluids. AB - To investigate early pulmonary responses to metalworking fluid exposure, we enrolled first-year machinist apprentices and apprentices in three other trades into a 2-yr longitudinal study. We obtained complete data for 82 machinists and 159 control subjects. Tests included respiratory questionnaires, spirometry, methacholine challenge, and allergy skin tests. Details on duration of exposure were collected by interview and 68 representative full shift personal samples for "total aerosol" were obtained from 13 shops (mean: 0.46 mg/m3, range: < 0.7 to 3.65 mg/m3). Machinists and control subjects did not differ at baseline. At follow-up, average change in bronchial responsiveness was double in machinists compared with control subjects (p = 0.05), and machinists were more likely to have developed new bronchial hyperresponsiveness (BHR) with asthmalike symptoms. In linear regression analysis, for predictors of methacholine slope, increased BHR was associated with duration of exposure to both synthetic and soluble metalworking fluids (p < 0.05); in logistic regression analysis, for predictors of BHR, only duration of exposure to synthetic fluids was a significant predictor. Results were not changed when workers with PC20 < 8 mg/ml at baseline were excluded. We conclude that exposure to water-based metalworking fluids (especially synthetic fluids) is associated with increasing BHR during the first 2 yr of exposure. PMID- 9872824 TI - Risk factors for pulmonary mycobacterial disease in South African gold miners. A case-control study. AB - Pulmonary mycobacterial disease is common in miners. Risk factors for nontuberculous pulmonary mycobacterial (NTM) disease and tuberculosis (TB) in gold miners were identified in a retrospective case-control study that included 206 NTM patients and 381 TB patients of known human immunodeficiency virus (HIV) status diagnosed between 1993 and 1996. A total of 180 HIV-tested trauma/surgical inpatients were selected as control patients. Both HIV infection (odds ratio [OR] 3.6 for NTM and 4.5 for TB patients) and higher grades of silicosis (OR 5.0 for NTM and 4.9 for TB patients) were significantly more common in NTM and TB patients than in control patients. HIV prevalence rose in the control and both case groups during the study period. The overall HIV prevalence was 13.1% in NTM patients, 14.2% in TB patients, and 5.6% in control patients. Previous TB (OR 9.6), premorbid focal radiological scarring (OR 7.4) and a dusty job at diagnosis (OR 2.4) were additional significant risk factors for NTM disease. These findings suggest that the historically high incidence of NTM disease in miners is largely attributable to chronic chest disease from silica dust inhalation and prior TB. HIV infection has recently become an additional risk factor for mycobacterial disease in miners and is likely to become increasingly important as the HIV epidemic progresses. PMID- 9872825 TI - Iron uptake promotes hyperoxic injury to alveolar macrophages. AB - Iron uptake by cells may increase the intracellular pool of prooxidant iron prior to storage of iron within ferritin. Because hyperoxia is toxic to alveolar macrophages (AM) via mechanisms involving oxidant stress, we hypothesized that iron uptake by AM might promote hyperoxia-induced injury. To assess this hypothesis, we cultured AM recovered from healthy volunteers under conditions of normoxia or hyperoxia (60% or 95% oxygen) in media of varying iron content, including control media (3 microM iron) and media supplemented with iron (FeCl3; total iron 10, 20, or 40 microM). AM injury was assessed by measuring release of lactate dehydrogenase (LDH), phagocytic activity for yeast, and cytosolic concentrations of calcium ([Ca2+]i) as determined by ratio image analysis of AM loaded with the fluorescent calcium probe indo-1. There was dose-dependent accumulation of iron and ferritin synthesis in AM exposed to iron-supplemented media. Exposure of AM to hyperoxia (60% and 95% oxygen, 18 h) in control media increased LDH release and impaired phagocytic activity for yeast; however, similar hyperoxic exposures in iron-supplemented media significantly increased the cells' LDH release and decreased phagocytosis. Exposure to 95% oxygen increased the [Ca2+]i of AM over 18 h, but similar exposure in iron-supplemented media induced greater increases in [Ca2+]i. As compared with exposure to normoxia, exposure to hyperoxia (60% and 95% oxygen) also decreased iron uptake and, to a greater extent, ferritin synthesis by AM in iron-supplemented media. These data suggest that: (1) iron uptake promotes hyperoxic injury to AM; and (2) hyperoxia impairs the capacity of AM to sequester iron in ferritin. PMID- 9872826 TI - Sniff nasal inspiratory pressure. Reference values in Caucasian children. AB - Like in adults, normal values of maximal inspiratory pressure (PImax) and maximal expiratory pressure (PEmax) span a large range in children, making interpretation of low values difficult. Recently, sniff nasal inspiratory pressure (Pnsn) was developed as a new noninvasive test of inspiratory muscle strength. In healthy adults, Pnsn is most often higher than PImax. The aim of this study was to establish reference values of Pnsn in children and to compare them with PImax. A group of 180 unselected healthy children age 6 to 17 yr was studied in a school setting. All had a forced vital capacity (FVC) > 80% of predicted and a ratio of forced expiratory volume in one second/forced vital capacity (FEV1/ FVC) > 85% of predicted. All maneuvers were performed in the sitting position. The Pnsn was measured using a catheter occluding one nostril during maximal sniffs performed through the contralateral nostril from FRC. The PImax was measured from FRC and residual volume, and PEmax from FRC and total lung capacity. All children were able to perform the Pnsn maneuver easily. Pnsn was 104 +/- 26 cm H2O in boys and 93 +/- 23 cm H2O in girls (p < 0.005). These values were similar to those previously measured in healthy adults. Pnsn correlated with age, weight, and height in boys, but not in girls. In both sexes, Pnsn was higher than PImax measured at the same lung volume (FRC) (p < 0. 0001). Pnsn was >= PImaxFRC in 73 of 93 boys and 79 of 87 girls. We conclude that Pnsn can be easily used to assess inspiratory muscle strength in children age 6 yr or more, providing values higher than PImax. Normal values are independent of age in girls, and can be predicted from age by a first-degree equation in boys. Being easy and noninvasive, Pnsn may prove useful to assess inspiratory muscle strength in children with neuromuscular disorders. PMID- 9872827 TI - Breathing during sleep in patients with nocturnal desaturation. AB - The mechanisms leading to hypoxemia during sleep in patients with respiratory failure remain poorly understood, with few studies providing a measure of minute ventilation (V I) during sleep. The aim of this study was to measure ventilation during sleep in patients with nocturnal desaturation secondary to different respiratory diseases. The 26 patients studied had diagnoses of chronic obstructive pulmonary disease (COPD) (n = 9), cystic fibrosis (CF) (n = 2), neuromusculoskeletal disease (n = 4), and obesity hypoventilation syndrome (OHS) (n = 11). Also reported are the results for seven normal subjects and seven patients with effectively treated obstructive sleep apnea (OSA) without desaturation during sleep. Ventilation was measured with a pneumotachograph attached to a nasal mask. In the treated patients with OSA and in the normal subjects, only minor alterations in V I were observed during sleep. In contrast, mean V I for the group with nocturnal desaturation decreased by 21% during non rapid-eye-movement (NREM) sleep and by 39% during rapid-eye-movement (REM) sleep as compared with wakefulness. This reduction was due mainly to a decrease in tidal volume (V T). Hypoventilation was most pronounced during REM sleep, irrespective of the underlying disease. These data indicate that hypoventilation may be the major factor leading to hypoxia during sleep, and that reversal of hypoventilation during sleep should be a major therapeutic strategy for these patients. PMID- 9872828 TI - Longitudinal decline in measured firefighter single-breath diffusing capacity of carbon monoxide values. A respiratory surveillance dilemma. AB - Seattle firefighters participate in a voluntary annual medical surveillance program including measurements of ventilatory capacity (FVC and FEV1) and single breath diffusing capacity of carbon monoxide (DLCO). From 1989 to 1996, average % predicted DLCO (Crapo) for all participating firefighters declined from 94.4% (95% confidence interval [CI]: 93.4% to 95.5%) to 87.3% (95% CI: 86.2% to 88.3%), with no significant change in average FVC or FEV1. A random-effects regression model based on data from 812 firefighters with at least two annual sets of DLCO measurements showed the expected associations between DLCO and age, height, gender, race, ventilatory capacity, and smoking. In addition, two important temporal changes were observed, including, for an average firefighter, a large mean decline in DLCO of -1.02 ml/min/mm Hg associated with year of measurement, and a relatively smaller decline of -0.006 ml/min/mm Hg associated with number of fires fought. Although the stability of ventilatory capacity over time is reassuring, the marked temporal decline in diffusing capacity among this population of firefighters raises issues of concern. Interpretation of the observed decline poses a dilemma in terms of the reliability and efficacy of diffusing capacity as a screening tool, in whether DLCO is subject to unacceptable technical variability or whether it might provide more sensitive detection of early adverse respiratory effects of smoke inhalation. PMID- 9872829 TI - Asthma, wheezy bronchitis, and atopy across two generations. AB - Although the prevalence of asthma has risen significantly during the last 30 yr, it is not clear whether this has occurred primarily in persons with a strong genetic predisposition to asthma and atopy or in other sections of the population. We have investigated outcomes in children of nuclear families selected through probands previously characterized by studies in 1964 and 1989 as having histories of persistent childhood onset atopic asthma, transient childhood wheezy bronchitis, and no respiratory symptoms or atopy. Children of wheezy bronchitic probands had a significantly better symptomatic outcome in adolescence, irrespective of the atopic status of the parent proband, than do children of either asthmatic or asymptomatic probands, suggesting that this may be a syndrome that shows familial aggregation and is distinct from asthma. Total serum IgE levels were significantly lower in children of nonatopic asymptomatic probands, including those with wheezing symptoms. In contrast children of nonatopic asymptomatic probands had an unexpectedly high prevalence of wheezing (33%), positive skin prick tests (56%), and positive specific serum IgE to common allergens (48%) that was similar to that found in children of atopic asthmatic probands. Our findings support the concept that wheezy bronchitis is a separate syndrome from atopic asthma. High total serum IgE levels within our population appear to be an important marker of genetic predisposition to atopy. Our data also suggest that much of the increase in asthma prevalence is associated with specific IgE sensitization and is occurring in persons previously considered to be at low risk of developing asthma or atopy. PMID- 9872830 TI - Variation in the arousal pattern after obstructive events in obstructive sleep apnea. AB - The relationship between the severity of obstructive sleep apnea (OSA) (measured by sleep study) and daytime sleepiness is poor. Variation in the degree of arousal accompanying obstructive respiratory events might help explain this poor correlation. Polysomnographic records from patients with OSA were reviewed in order to extract representative examples of apneas and hypopneas (in 10 patients), as well as events both supine and decubitus (in 12 patients). The EEG accompanying each obstructive event was processed with a neural network technique to describe sleep depth on a second-by-second basis. The lengths of any visually evident microarousals were also measured manually. There was considerable interindividual variation in the degree of sleep disturbance using the neural network technique (p < 0.005), but not using the lengths of the visually scored microarousals (p = 0.6). The arousals accompanying apneic events caused greater variability in sleep depth quantified using the neural network technique (p = 0.03), and also lasted longer based on the visual scoring (mean, 12.6; SD, 1.7 s) than the hypopneic events (mean, 9.9; SD, 2.4 s; p = 0.02). There were no significant differences between events occurring supine versus decubitus with either technique (p = 0.7). These differences in arousal magnitude may explain some of the poor correlations between conventional measures of sleep apnea severity and daytime sleepiness. PMID- 9872831 TI - Influence of shift work and host factors on endotoxin-related acute peak flow changes. AB - There is limited knowledge about which internal and external factors influence respiratory effects of occupational endotoxin exposure. Relationships between endotoxin exposure, work-related respiratory symptoms, and acute peak flow changes were studied in 97 shift workers in the potato processing industry. For each worker, across-shift peak flow changes were determined for morning, afternoon, and night shifts. A higher endotoxin exposure was associated with an increased prevalence of work-related symptoms, a smaller peak expiratory flow (PEF) increase across the morning shift, and a larger PEF decrease across afternoon and night shifts. The largest effects occurred during the afternoon shift. No consistent differences between smokers and nonsmokers, and between atopics and nonatopics were observed. We conclude that endotoxin exposure in the potato processing industry is related to across-shift peak flow changes and the occurrence of work-related respiratory symptoms. The host factors smoking and atopy are not important confounding or effect-modifying factors in these relationships. In respiratory health studies among shift workers, it is important to investigate all work shifts before drawing definitive conclusions about exposure-response relationships. PMID- 9872832 TI - Cytokine levels correlate with a radiologic score in active pulmonary tuberculosis. AB - Pulmonary tuberculosis is an infectious disease caused by Mycobacterium tuberculosis. This microorganism is capable of inducing a delayed hypersensitivity reaction in the lung, with subsequent expression of the disease. This reaction depends on the presence of different cytokines that exert specific functions. The aim of this study was to evaluate the presence and the concentrations of nine different modulators in bronchoalveolar lavage fluid (BALF). For this purpose, 15 patients with active pulmonary tuberculosis were enrolled at the time of diagnosis, prior to institution of antituberculous therapy. All the patients demonstrated M. tuberculosis in the sputum, and their disease extention was defined by high-resolution computed tomography (HRCT) using a score which included the presence of six findings: miliary nodules, nodules < 10 mm, consolidation, ground glass, cavity and bronchial wall thickening. This score was more sensitive than an equivalent score calculated on the basis of chest radiology. HRCT score was calculated for each area of the two lungs in order to define the more and the less affected lung for each patient. The bronchoalveolar lavage (BAL) was performed in the more affected area for each lung. The HRCT total score for each washed area ranged between 1 and 15, and showed more significant differences between the more and less affected lungs (p = 0.0004) than those obtained with the individual radiologic findings (p ranged between 0.60 and 0. 004). The BAL concentrations of the nine cytokines evaluated for the more and less affected lungs were compared: interleukin-6 (IL-6), IL-8, IL-12, tumor necrosis factor-alpha (TNF-alpha), and interferon gamma (IFN-gamma) showed significant differences (p ranged between 0. 016 and 0.0007). In addition, each cytokine concentration was correlated with the HRCT score. Significant correlations were found with IL-12, IL-6, IL-8, IL-2, and TNF-alpha. The correlations between cytokines and HRCT total score were better than those observed with the individual radiologic findings. A correlation matrix for the different cytokines evaluated one against each other, has also been added to show common behavior of these modulators. A similar analysis was also performed for the radiologic abnormalities. PMID- 9872833 TI - Pharyngeal critical pressure in patients with obstructive sleep apnea syndrome. Clinical implications. AB - Current evidence suggests that patients with obstructive sleep apnea (OSA) may have greater pharyngeal critical pressure (Pcrit), which reflects the increase in upper airway collapsibility. The contribution of Pcrit to the severity of OSA and to the efficacious continuous positive pressure (nCPAPeff) therapy has never been extensively described and no data are available about the interaction of Pcrit, age, and anthropometric variables. To determine the relationship between Pcrit, severity of the disease, nCPAPeff, and anthropometric variables we measured Pcrit in a group of 106 patients with OSA. Pharyngeal critical pressure was derived from the relationship between maximal inspiratory flow and nasal pressure, Pcrit representing the extrapolated pressure at zero flow. Upper airway resistance (Rus) was determined as the reciprocal of the slope (DeltaPn/DeltaVImax cm H2O/L/s) in the regression equation. In a subgroup of 68 patients, during the diagnostic night, we measured as indices of respiratory effort, the maximal inspiratory esophageal pressure (Pes) at the end of apnea (Pesmax), the overall increase from the minimum to the maximum (DeltaPes), and the rate of increase of Pes during apnea (RPes). As a group, the mean Pcrit was 2.09 +/- 0.1 cm H2O (range, 0 to 4.5) and the mean Rus was 11.1 +/- 0.5 cm H2O/L/s. Although men have greater Pcrit, pharyngeal collapsibility was influenced neither by neck size nor by body mass index (BMI). Although there was a significant relationship between Pcrit and apnea plus hypopnea index (AHI) (r = 0.23, p = 0.02), neck circumference was the stronger predictor of apnea frequency, with Pcrit contributing only to the 3% of the variance. In the group of patients as a whole, a model including AHI, BMI, Rus, and Pcrit explained the 36% of the variance in nCPAPeff, with a greater contribution of AHI, Pcrit accounting for only 3% of the variation. In patients for whom the measure of respiratory effort was obtained, 42% of the variance in nCPAPeff was explained by RPes (33%) and BMI. From these results we conclude that Pcrit alone does not yield a diagnostically accurate estimation of OSA severity and nCPAPeff. Although individual collapsibility may predispose to pharyngeal collapse, upper airway occlusion may require the combination of several factors, including obesity, upper airway structure, and abnormalities in muscle control. PMID- 9872834 TI - Predictive factors of hospitalization for acute exacerbation in a series of 64 patients with chronic obstructive pulmonary disease. AB - Hospitalizations for acute exacerbation in patients with chronic obstructive pulmonary disease (COPD) have a great impact on health care expenditure. The aim of this study was to look at predictive factors of hospitalization for acute exacerbation in a group of patients with moderate to severe COPD. During the year 1994, we included 64 patients with COPD in this study. At inclusion, the patients being in a stable state, we performed a complete evaluation of their clinical, spirometric, gasometric, and pulmonary hemodynamic characteristics. All patients were followed during a period of at least 2.5 yr. We recorded the intervals free of hospitalization for exacerbation and realized an analysis of the proportional hazards not to be hospitalized using the Kaplan-Meier method. Univariate analysis using the log-rank test showed that the risk of being hospitalized was significantly increased in patients with COPD with a low body mass index (BMI <= 20 kg/m2, p = 0.015) and in patients with a limited 6-min walk distance (<= 367 m, p = 0. 045). But above all, the risk of hospitalization for acute exacerbation was significantly increased by gas exchange impairment and pulmonary hemodynamic worsening: PaO2 <= 65 mm Hg versus PaO2 > 65 mm Hg, p = 0.005; PaCO2 > 44 mm Hg versus PaCO2 <= 44 mm Hg, p = 0.005; and mean pulmonary artery pressure ( Ppa) at rest > 18 mm Hg versus Ppa <= 18 mm Hg, p = 0.0008. Neither age, nor the association of one or more comorbidities with COPD, nor the smoking habits had a significant impact on the risk of hospitalization in our study. Multivariate analysis showed that only PaCO2 and Ppa were independently related to the risk of hospitalization for acute exacerbation of COPD. We conclude that chronic hypercapnic respiratory insufficiency and pulmonary hypertension are predictive factors of hospitalization for acute exacerbation in COPD patients. PMID- 9872835 TI - Guinea pig airway hyperresponsiveness induced by blockade of the angiotensin II type 1 receptor. Role for endogenous nitric oxide. AB - Losartan is the first angiotensin II type 1 (AT1) receptor antagonist to become available for the treatment of hypertension. However, recent reports have revealed several cases of losartan-induced bronchoconstriction. We investigated to determine the mechanism of losartan-induced bronchoconstriction, considering in particular the involvement of endogenous nitric oxide (NO). In this study, we examined the effects of losartan on airway obstruction and endogenous NO production using anesthetized guinea pigs and cultured airway epithelial cells. Five minutes after administration of angiotensin II (Ang II), the bronchoconstriction induced by acetylcholine was not changed. In contrast, Ang II in the presence of losartan caused a significant increase in the acetylcholine responsiveness. Pretreatment with L-N omega-nitroarginine-methylester (L-NAME) potentiated acetylcholine-induced bronchoconstriction 5 min after administration of Ang II, and L-arginine reversed this action of L-NAME on the acetylcholine responsiveness. Moreover, Ang II administration increased NO concentration in expired air (12.5 +/- 1.5 ppb for saline, 40 +/- 5 ppb for Ang II, p < 0.01), and losartan significantly inhibited Ang II-stimulated NO release (20 +/- 3.5 ppb) from guinea pig airway. In cultured airway epithelial cells, Ang II also increased NO release (160 +/- 25 nM), and the effect of this Ang II-induced NO release was significantly inhibited by pretreatment with losartan (25 +/- 8 nM, p < 0.01). These findings suggest that losartan-induced bronchoconstriction may result from inhibition of endogenous NO release in the airway. PMID- 9872836 TI - Airway and lung tissue mechanics in asthma. Effects of albuterol. AB - We examined the partitioning of total lung resistance (RL) into airway resistance (Raw) and tissue resistance (Rti) in patients with mild to moderate asthma (baseline FEV1, 54 to 91% of predicted) before and after albuterol inhalation. An optimal ventilator waveform was used to measure RL and lung elastance (EL) in 21 asthmatics from approximately 0.1 to 8 Hz during tidal excursions. Analysis of the RL and EL provided separate estimates of airway and lung tissue properties. Eleven subjects, classified as Type A asthmatics, displayed slightly elevated RL but normal EL. Their data were well described with a model consisting of homogeneous airways leading to viscoelastic tissues before and after albuterol. The other 10 subjects, classified as Type B asthmatics, demonstrated highly elevated RL and an EL that became highly elevated at frequencies above 2 Hz. These subjects required the inclusion of an airway wall compliance in the model prealbuterol but not postalbuterol. This suggests that the Type B subjects were experiencing pronounced constriction in the periphery of the lung, resulting in shunting of flow into the airway walls. Spirometric data were consistent with higher constriction in Type B subjects. Both groups demonstrated significant (p < 0.05) decreases in Raw and tissue damping after albuterol, but tissue elastance decreased only in the Type B group. The percent contributions of Raw and Rti to RL were similar in both groups and did not change after albuterol. We conclude that in asthma, Raw comprises the majority (> 70%) of RL at breathing frequencies. The relative contributions of Raw and Rti to RL appear to be independent of the degree of smooth muscle constriction. PMID- 9872837 TI - Spirometric reference values from a sample of the general U.S. population. AB - Spirometric reference values for Caucasians, African-Americans, and Mexican Americans 8 to 80 yr of age were developed from 7,429 asymptomatic, lifelong nonsmoking participants in the third National Health and Nutrition Examination Survey (NHANES III). Spirometry examinations followed the 1987 American Thoracic Society recommendations, and the quality of the data was continuously monitored and maintained. Caucasian subjects had higher mean FVC and FEV1 values than did Mexican-American and African-American subjects across the entire age range. However, Caucasian and Mexican-American subjects had similar FVC and FEV1 values with respect to height, and African-American subjects had lower values. These differences may be partially due to differences in body build: observed Mexican Americans were shorter than Caucasian subjects of the same age, and African Americans on average have a smaller trunk:leg ratio than do Caucasians. Reference values and lower limits of normal were derived using a piecewise polynomial model with age and height as predictors. These reference values encompass a wide age range for three race/ethnic groups and should prove useful for diagnostic and research purposes. PMID- 9872838 TI - Bacterial colonization patterns in mechanically ventilated patients with traumatic and medical head injury. Incidence, risk factors, and association with ventilator-associated pneumonia. AB - We prospectively evaluated the relation of upper airway, lower airway, and gastric colonization patterns with the development of pneumonia and its etiology in 48 patients with surgical (n = 25) and medical (n = 23) head injury. Initial colonization was assessed by cultures of nasal and pharyngeal swabs, tracheobronchial aspirates, gastric juice, and bronchoscopically retrieved protected specimen brush. Follow-up colonization was determined until the end points extubation, suspected ventilator-associated pneumonia (VAP), or death. The initial colonization rate at any site at ICU admission was 39/47 (83%). It mainly accounted for Group I pathogens (Streptococcus pneumoniae, Staphylococcus aureus, Hemophilus influenzae) of the upper and lower airways. At follow-up, colonization rates with Group II pathogens (Gram-negative enteric bacilli and Pseudomonas spp.) increased significantly. The high initial bacterial load with Group I pathogens of the upper airways and trachea decreased during Days 2 to 4, whereas that of Group II pathogens increased. Upper airway colonization was an independent predictor of follow-up tracheobronchial colonization (odds ratio [OR], 9.9; 95% confidence interval [CI], 1.8 to 56.3 for initial colonization with Group I pathogens; OR, 23.9; 95% CI, 3.8 to 153.3 for follow-up colonization with Group II pathogens). Previous (short-term) antibiotics had a protective effect against colonization with Group I pathogens of the lower respiratory tract (OR, 0.2; 95% CI, 0.05 to 0.86), but they were a risk factor for colonization with Group II pathogens (OR, 6.1; 95% CI, 1.3 to 29). Initial tracheobronchial colonization with Group I pathogens was associated with a higher probability of early onset pneumonia (OR, 4. 1; 95% CI, 0.7 to 23.3), whereas prolonged antibiotic treatment (> 24 h) independently predicted late-onset pneumonia (OR, 9.2; 95% CI, 1.7 to 51.3). We conclude that patients with head injury are colonized in the airways mainly by Group I pathogens early in the evolution of illness. The upper airways represent the main reservoir for subsequent lower airway colonization with Group I pathogens. Previous (short-term) antibiotic treatment is protective against initial tracheobronchial colonization with Group I pathogens, but it represents a risk factor for subsequent lower airway colonization by Group II pathogens. PMID- 9872839 TI - Mixed hematopoietic chimerism induces donor-specific tolerance for lung allografts in rodents. AB - Mixed hematopoietic chimerism is a state in which bone marrow hematopoietic stem cells from two genetically different animals coexist. We investigated whether mixed hematopoietic chimerism, resulting from the transplantation of host and donor bone marrow into a lethally irradiated rat, would confer donor-specific tolerance to lung allografts. Recipient rats (Fisher or or Wistar Furth [WF]) were irradiated (1,100 cGy) and reconstituted with a mixture of T-cell-depleted syngeneic plus allogeneic bone marrow. After mixed chimerism was documented by the presence of donor- and host-derived cells in the peripheral blood 4 wk after bone marrow reconstitution, mixed chimeras underwent orthotopic left lung transplantation with donor-specific and third-party lung allografts. No immunosuppressive agents were administered after lung transplantation. All donor specific lung allografts were accepted by mixed chimeras (n = 40), while all third-party grafts (n = 7) were rejected within 10 d, a time course similar to that for grafts transplanted into naive recipients (n = 14). Radiation control recipients (n = 7) who did not develop mixed chimerism because the donor bone marrow had failed to engraft, also rejected donor-specific grafts within 10 d. We conclude that mixed hematopoietic chimerism induces donor-specific transplantation tolerance to lung allografts. PMID- 9872840 TI - Role of actin depolymerization in the surfactant secretory response of alveolar epithelial type II cells. AB - Alveolar epithelial type II cells (AET2) respond with exocytosis of surfactant containing lamellar bodies to stimulation with mechanical stretch and secretagogues, a process that is fundamental for maintaining alveolar stability and lung gas exchange. In the present study in cultured rat AET2, we employed botulinum C2 toxin, a binary toxin which ADP ribosylates nonmuscle G-actin, as a specific tool to probe the role of the actin microfilament system in the surfactant secretory process. Incubation of AET2 with C2 toxin caused a dose dependent decay of the cellular F-actin content to a minimum of 20% of baseline, concomitant with an increase in monomeric actin. In parallel, a significant augmentation of baseline surfactant secretion up to twofold elevated levels above control was noted, as assessed by the release of prelabeled phosphatidylcholine. Pretreatment with phalloidin, which stabilized F-actin and reduced the level of G actin, prevented the C2 toxin-elicited enhancement of baseline surfactant secretion. Even low C2 toxin concentrations, resulting in a reduction of total cellular F-actin content of approximately 10%, sufficed to augment secretagogue (ATP) and, more impressively, mechanical stress elicited an increase in surfactant secretion; the response to the biophysical challenge more than doubled. When investigated in the absence of toxin, different secretagogues (ATP, phorbol ester, betamimetics) caused a rapid-onset, transient reduction of F-actin in the range between 15 and 25% as a consistent part of their secretory response pattern. These data suggest that the state of actin polymerization is intimately linked to the exocytosis process underlying surfactant secretion in AET2. Microfilament system-related compartmentalization effects and/or or the impact of the state of actin assembly on signaling events may be considered as underlying events. PMID- 9872841 TI - Peripheral neuropathy in sleep apnea. A tissue marker of the severity of nocturnal desaturation. AB - Because chronic obstructive pulmonary disease (COPD) is well known to induce peripheral neuropathy and resistance to ischemic nerve conduction failure (RICF), we performed a case-control study examining peripheral nerve function during ischemia in 17 patients with severe obstructive sleep apnea (OSA) without daytime hypoxemia and 10 control subjects. Median nerve conduction was studied before, during, and after a 30-min period of ischemia. Preischemic sensory and mixed nerve potential amplitudes and sensory conduction velocity were lower in OSA patients than in control subjects despite higher supramaximal stimulation. During ischemia, seven OSA patients manifested RICF (OSA-RICF), whereas both the other 10 patients, who were nonresistant to ischemic conduction failure (OSA-NR), and control subjects did not. OSA-RICF patients had the lowest initial nerve potential amplitude, whereas OSA-NR patients had a response intermediate between that of control subjects and OSA-RICF patients. OSA-RICF patients had a lower mean nocturnal SaO2 and a higher body mass index (BMI) and duration of SaO2 < 70% than did OSA-NR patients. Seven patients (four OSA-RICF and three OSA-NR) were reevaluated after at least 2 mo of treatment with nasal continuous positive airway pressure (nCPAP). RICF disappeared in all OSA-RICF patients, whereas preischemic nerve conduction parameters were unchanged in both OSA-RICF and OSA NR patients. Thus OSA patients have peripheral nerve dysfunction whose severity is partly related to the level of nocturnal hypoxemia. Abnormal preischemic nerve conduction suggests axonal lesions, whereas RICF, which appears to be a sensitive but nonspecific tissue marker of the severity of hypoxemia, may result from adaptative mechanisms. PMID- 9872842 TI - Lovastatin induces fibroblast apoptosis in vitro and in vivo. A possible therapy for fibroproliferative disorders. AB - Diseases associated with pathological fibroproliferation represent a major cause of morbidity and mortality. Despite the importance of this class of disorders, current therapy is of limited value, and no therapy is available to reduce the fibroblast population size within existing fibrotic lesions. In this regard, constitutive expression of growth-promoting genes can sensitize cells to undergo apoptosis. Studies in our laboratory have demonstrated that lovastatin potently induces apoptosis in fibroblasts constitutively expressing Myc, and that lung fibroblasts isolated from fibrotic lesions constitutively express growth promoting genes. In this study, we sought to determine if nontransformed lung fibroblasts would manifest susceptibility to lovastatin-induced apoptosis similar to that observed in fibroblasts ectopically expressing Myc. Here we show that clinically achievable concentrations of lovastatin induce apoptosis in normal and fibrotic lung fibroblasts in vitro, as evidenced by acridine orange staining, terminal transferase nick end translation (TUNEL), and DNA laddering. Apoptosis of human lung fibroblasts was dose- and time-dependent, and blocked by exogenous mevalonic acid. Furthermore, apoptosis was associated with decreased levels of mature Ras, a molecule directly implicated in fibroblast rescue from apoptosis. The ability of lovastatin to induce fibroblast apoptosis in vivo was examined using a guinea pig wound chamber model. Lovastatin (5 microM, 8 d) reduced granulation tissue formation in the wound chambers by 64.7%, with associated ultrastructural evidence of fibroblast apoptosis. These findings support further study of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors as potential therapy for patients with fibroproliferative disorders. PMID- 9872843 TI - Lymphocyte and eosinophil influx into alveolar tissue in nocturnal asthma. AB - We have shown in nocturnal asthma that alveolar tissue eosinophils are increased at night as compared with the proximal airway, and that they correlate with the overnight decrement in lung function. As the CD4+ cell is thought to be the principal orchestrating cell in eosinophil recruitment, we evaluated its presence in the proximal and distal airways in nocturnal asthma. Eleven patients with nocturnal asthma (NA) and 10 patients with non-nocturnal asthma (NNA) underwent two bronchoscopies with proximal airway endobronchial and distal alveolar tissue transbronchial biopsy in a random order at 4:00 P.M. and at 4:00 A.M. separated by 1 wk. Immunohistochemical staining and morphometric analysis were used to determine the number of CD3+, CD4+, and CD8+ cells and EG2+ eosinophils per mm2 in the epithelium, lamina propria, and alveolar tissue. At 4:00 A.M., the NA group had a significantly greater number of CD4+ cells in the alveolar tissue than the NNA group (9.8 cells/ mm2 [5.6-30.8, interquartile (IQ)] versus 1.5 cells/mm2 [0-6. 3, IQ], p = 0.04). Within the NA group, there were significantly greater numbers of CD3+, CD4+, CD8+, and EG2+ cells in the proximal airway lamina propria than in the distal airway at both 4:00 P.M. and 4:00 A.M. There were no differences within the epithelium between the groups at either time point. Only alveolar tissue, not airway tissue, CD4+ cells correlated inversely with the percentage predicted FEV1 at 4:00 A.M. (r = -0.68, p = 0.0018) and positively with the number of alveolar tissue EG2+ cells (r = 0.66, p = 0.01). These findings suggest that the CD4+ lymphocyte is increased in the alveolar tissue at night in nocturnal asthma as compared with non-nocturnal asthma. PMID- 9872845 TI - Surfactant subtype conversion is related to loss of surfactant apoprotein B and surface activity in large surfactant aggregates. Experimental and clinical studies. AB - Conversion of the highly surface-active subtype of pulmonary surfactant known as large surfactant aggregates (LA) to small aggregates (SA) with poor surface activity has recently been shown to occur upon cyclic changes of the air-liquid interface area in vitro. By subjecting pooled rabbit bronchoalveolar lavage fluid (BALF) to this maneuver, we found that conversion of LA to SA was accompanied by a marked decline in the ability of the remaining LA fraction to reduce surface tension by adsorption and during film compression on a pulsating bubble surfactometer. SA obtained by centrifugation of noncycled rabbit BALF had a similar phospholipid (PL) but different neutral lipid (NL) composition than did the LA. Upon cycling, the increased formation of SA obliterated this difference. No substantial difference in the PL, NL, or fatty acid profile of LA was noted before and after cycling. In contrast, the content of surfactant apoprotein-B (SP B) in the LA decreased dramatically to nearly undetectable levels during the cycling maneuver, and this decline in SP-B content was closely correlated with the decrease in proportional appearance of LA and loss of surface activity of this fraction. Reconstitution of LA with intact SP-B after cycling virtually fully restored the surface activity of this surfactant subtype. When comparing lavage samples from adults with acute respiratory distress syndrome (ARDS; n = 10) with samples from healthy controls (n = 11), we noted a marked reduction of SP-B in the LA fraction. There was a significant correlation between the SP-B content of the LA fraction and the relative percentage of LA in BALF or the lower surface activity of this surfactant subtype. We conclude that an SP-B-related loss of LA integrity and function may substantially contribute to the decline of this surfactant subtype and the loss of its surface activity during cycling in vitro and in clinical ARDS. PMID- 9872844 TI - Bronchoprotector properties of calcitonin gene-related peptide in guinea pig and human airways. Effect of pulmonary inflammation. AB - Calcitonin gene-related peptide (CGRP), a neuropeptide released from sensory nerves during axonal reflexes, has strong bronchoprotector properties in rat isolated airways. In this study, we examined this ability of CGRP to prevent agonist-induced contraction in guinea pig and human airways and determined whether inflammatory reaction affects its function. CGRP administered intravenously (0.38 to 114 microgram/kg) in anesthesized guinea pig had no effect per se on airway resistance but caused a dose-related inhibition of substance P (SP; 13.5 microgram/kg)-induced bronchoconstriction (60% at 114 microgram/kg). Similarly, CGRP (10(-)9 to 10(-)6 M) prevented in a concentration-dependent manner the contraction elicited by SP (5 x 10(-)8 M) in guinea pig isolated main bronchi and parenchymal strips, the inhibition caused by CGRP being more pronounced in distal than in proximal airways (47 and 20%, respectively, at 10( )6 M). The breaking effect of CGRP on SP-induced constriction was however significantly reduced (p < 0.05) in guinea pig actively sensitized to ovalbumin (OA) and the loss in its potency was of similar magnitude (> 40%) whether it was administered in vivo or in vitro. A same phenomenon was observed in human isolated peripheral bronchi. CGRP (10(-)6 M) reduced by more than 75% the extent of the contraction evoked by 10(-)6 M of carbamylcholine and its protector effect was totally abolished in bronchi showing clear morphological manifestation of inflammatory reaction. It is concluded that CGRP acts as a potent bronchoprotector agent on both guinea pig and human airways but its ability to limit the extent of airway responsiveness is strongly impaired in inflammatory conditions. PMID- 9872846 TI - Role of nitric oxide in sepsis-associated pulmonary edema. AB - Transient pulmonary hypertension after inhibition of nitric oxide synthase (NOS) does not alter pulmonary reflection coefficients or lymph flows in endotoxemic sheep. To test the effects of persistent pulmonary hypertension induced by N omega-nitro-L-arginine methylester (L-NAME) and of inhaled NO on pulmonary edema, 18 sheep (three groups) were chronically instrumented with pulmonary artery catheters, femoral arterial fiberoptic thermistor catheters, and tracheostomy. The awake, spontaneously breathing animals received Salmonella typhi endotoxin (lipopolysaccharide; LPS) (10 ng/kg/ min) for 28 h. After 24 h, an airflow of 6 L/min was delivered through the tracheostomy. One group of animals (L-NAME/air) received L-NAME intravenously (25 mg/kg + 5 mg/kg/h) and breathed air. The second group (L-NAME/NO) was given L-NAME and NO (40 ppm) was added to the airflow. The third group was given NaCl 0.9% and breathed air (NaCl/air). Extravascular lung water was measured through the double-indicator dilution technique. Endotoxemia caused pulmonary edema, which was aggravated by L-NAME. Breathing of NO normalized pulmonary artery pressure (Ppa) and ameliorated pulmonary edema. Inhalation of NO may therefore be a therapeutic option for pulmonary edema associated with pulmonary hypertension. PMID- 9872847 TI - Unopposed neutrophil elastase in bronchoalveolar lavage from transplant recipients with cystic fibrosis. AB - Large numbers of neutrophils with unopposed neutrophil elastase (NE) proteolytic activity are found in lower respiratory tract secretions from most patients with advanced cystic fibrosis (CF). To determine whether antielastase defenses may be overwhelmed in epithelial lining fluid after lung transplantation, we measured NE activity (cleavage of the specific substrate, MeO-Suc-Ala-Ala-Pro-Val-pNA) in bronchoalveolar lavage fluids (BALF) obtained for surveillance or diagnostic purposes at various intervals (1 mo to 7 yr after transplantation) from 52 recipients who had undergone double or bilateral lung transplantation for end stage CF. Unopposed NE activity was found in BALF from 14 recipients, most of whom also had >= 10(5) colony forming units (cfu) of Pseudomonas aeruginosa in BALF. Ten of the 14 recipients with unopposed NE in bronchoalveolar lavage (BAL) had developed obliterative bronchiolitis (OB), but only 8 of the 38 subjects without unopposed NE activity had OB (p = 0. 002; Fisher exact test). We conclude that antiprotease defenses in lower respiratory tract secretions of CF patients receiving lung allografts are sufficient in the majority of patients to prevent unopposed NE activity. However, the presence of unopposed NE activity in BAL from lung allografts of patients with CF is associated with progressive, irreversible OB and graft failure. PMID- 9872848 TI - Effect of natural grass pollen exposure on exhaled nitric oxide in asthmatic children. AB - Exhaled nitiric oxide (NO) is increased in exhaled breath of asthmatic patients. The aim of this study was to investigate the longitudinal changes of exhaled NO outside and during the pollen season in pollen-allergic asthmatic children. Twenty-one children (age 6 to 16 yr), with a seasonal allergic asthma sensitive to grass pollen, underwent measurements of exhaled NO and pulmonary function before (March), during (May), and after (November) the pollen season. Exhaled NO was measured by a tidal breathing method with a chemiluminescence analyzer and NO steady-state levels were recorded. The timing of the measurements during the pollen season was based on the atmospheric pollen count. Exhaled NO values of asthmatic children were compared with those of 21 sex- and age-matched healthy children. Pulmonary function and symptoms of asthma were also evaluated at each visit. The mean value of exhaled NO before the grass season was 12.7 +/- 5.1 ppb (mean +/- SD), significantly higher when compared with controls (7.8 +/- 2.7 ppb, p < 0.001). In the pollen season there was a significant (p < 0.001) twofold increase in exhaled NO (21.4 +/- 7.6 ppb) that, after the season, returned to values similar (12.8 +/- 5.8 ppb, p = NS) to those found before the season. There were no significant changes in FEV1 before and during the season (98.6% predicted versus 101% predicted, p = NS). We conclude that natural allergen exposure is related to an increase of exhaled NO in asthmatic grass pollen-allergic children even in absence of significant changes in airways function. We speculate that measurement of exhaled NO could be a sensitive noninvasive marker of asthma disease activity. PMID- 9872849 TI - L- and P-selectin and CD11/CD18 in intracapillary neutrophil sequestration in rabbit lungs. AB - Infusion of complement fragments induces rapid sequestration of neutrophils within pulmonary capillaries. This study examined the mechanisms through which this sequestration occurs, as well as the effect of complement fragments on the expression of L-selectin and CD11/CD18 using ultrastructural immunohistochemistry. Studies using anti-P-selectin antibodies, fucoidin, L selectin-depleted neutrophils, and anti-CD18 antibodies showed that selectins and CD18 were not required for neutrophil sequestration. However, maintaining the sequestered neutrophils within the pulmonary capillaries required both L-selectin and CD11/CD18. Neutrophils in the pulmonary capillaries of rabbits given complement fragments expressed 72% less L-selectin and 98% more CD11/CD18 than did those in rabbits given saline. Shedding of L-selectin occurred preferentially from the microvillar processes of the plasma membrane rather than from the flat intervening regions. About 28% of L-selectin still remained on intracapillary neutrophil membranes after 15 min and was likely available for binding. Shedding of L-selectin appeared slower in vivo than in vitro. These studies indicate that neutrophil sequestration induced by complement fragments requires at least two sequential steps, one that does not require recognized adhesion molecules followed by a second that requires L-selectin and CD11/ CD18. PMID- 9872850 TI - A simple automated method for measuring pressure-volume curves during mechanical ventilation. AB - Measurement of respiratory compliance is advocated for assessing the severity of acute respiratory failure (ARF). Recently, the administration of an automated constant flow of 15 L/min was proposed as a method easier to implement at the bedside than supersyringe or inspiratory occlusions methods. However, pressure volume (P-V) curves were shifted to the right because of the resistive properties of the respiratory system. The aim of this study was to compare the P-V curves obtained using two constant flows-3 and 9 L/min-during volume-controlled mechanical ventilation with those obtained with the supersyringe and the inspiratory occlusions methods. Fourteen paralyzed patients with ARF were studied. The supersyringe and the inspiratory occlusions methods were performed according to usual recommendations. The new automated method was performed during volume-controlled mechanical ventilation by setting the inspiratory:expiratory ratio at 80%, the respiratory frequency at 5 breaths/min, and the tidal volume at 500 or 1,500 ml. These peculiar ventilatory settings were equivalent to administering a constant flow of 3 or 9 L/min during a 9.6-s inspiration. Esophageal and airway pressures were recorded. P-V curves obtained by the 3-L/min constant-flow method were identical to those obtained by the reference methods, whereas the P-V curve obtained by the 9-L/min constant flow was slightly shifted to the right. The slopes of the P-V curves and the lower inflection points were not different between all methods, indicating that the resistive component induced by administering a constant flow equal to or less than 9 L/min is not of clinical relevance. Because the 3-L/min constant-flow method is not artifacted by the resistive properties of the respiratory system and does not require any other equipment than a ventilator, it is an easy-to-implement, inexpensive, safe, and reliable method for measuring the thoracopulmonary P-V curve at the bedside. PMID- 9872851 TI - Acute treatment with corticosteroids decreases IGF-1 and IGF-2 expression in the rat diaphragm and gastrocnemius. AB - Massive doses of methylprednisolone (M) or triamcinolone (T) induced diaphragmatic type IIx/b atrophy, resulting in a leftward shift of the force frequency curve in rats (). To examine the role of insulin-like growth factors (IGFs) in these changes, IGF mRNA content was measured in costal diaphragm, gastrocnemius, and liver removed from 32 rats treated daily during 5 d either with saline (control, C and pair-fed, PF), M, or T (80 mg/kg). Blood samples were taken to measure IGF-1 serum levels. RNA levels were measured by Northern and dot blots after hybridization with rat IGF-1 or IGF-2 cDNA probes labeled with alpha 32P. Compared with C (845 +/- 128 ng/ml), IGF-1 serum levels were significantly decreased in M (699 +/- 90 ng/ml, p < 0.001 versus C) and PF animals (505 +/- 33 ng/ml, p < 0.001 versus others) and even more so, in T-treated animals (273 +/- 134 ng/ml, p < 0.001 versus others). Along the same lines, IGF-1 expression in the liver was depressed after corticosteroid treatment and in PF, whereas IGF-2 mRNA content remained unchanged. Compared with C, the relative expression of IGF 1 mRNA in the diaphragm was depressed by 44% and 69% in the M and T groups, respectively (p < 0.0001 versus C), while it was unchanged in PF animals. In the gastrocnemius, IGF-1 expression was reduced after M and T (-51% and -59%, respectively, p < 0.0001 versus C) as well as in PF animals (-40%, p < 0.001 versus C). For IGF-2, a similar pattern of expression was found in the diaphragm and the gastrocnemius. Indeed, IGF-2 mRNA tended to decrease in corticosteroid treated rats (NS) whereas it was unchanged in PF rats. We conclude that decreased IGF expression after corticosteroid treatment was similar in diaphragm and gastrocnemius and may be responsible for the diaphragmatic changes observed after steroid treatment. PMID- 9872852 TI - Clinical examination reliably detects intrinsic positive end-expiratory pressure in critically ill, mechanically ventilated patients. AB - Critically ill patients requiring mechanical ventilation often develop intrinsic positive end-expiratory pressure (PEEPi). Methods for its detection include an expiratory flow waveform display (not always available), an esophageal pressure transducer (invasive), or a relaxed or paralyzed patient. We sought to determine the accuracy of clinical examination for detecting PEEPi. Examiners blinded to waveform analysis assessed patients for the presence of PEEPi by inspection/palpation and auscultation. If either inspection/palpation or auscultation demonstrated PEEPi, it was said to be present by clinical exam. Clinicians with various levels of experience (attending, resident, student) made 503 observations of 71 patients. Sensitivity (SENS), specificity (SPEC), positive predictive value (PPV), negative predictive value (NPV), and likelihood ratios were determined for inspection/palpation, auscultation, and clinical exam. PEEPi was present during 69.8% of observations. SENS, SPEC, and PPV of clinical exam were 0.72, 0.91, and 0.95 respectively for the examiners as a whole. Likelihood ratio for PEEPi detection by clinical exam was 8.35. Attending intensivists displayed SPEC and PPV of 1.0. NPV was only 0.58 (likelihood ratio 0.31). We conclude that the clinical exam is very good for detecting PEEPi at all experience levels; and further, that the clinical exam is only modestly useful for ruling out PEEPi, therefore, other tests should be used if PEEPi is not detected by clinical exam. PMID- 9872853 TI - A tuberculin screening and isoniazid preventive therapy program in an inner-city population. AB - As tuberculosis transmission decreases, case rates decline and an increasing proportion of cases arises from the pool of persons with latent infection. Elimination of tuberculosis will require preventing disease from developing in infected persons. From 1994 to 1996 the Atlanta TB Prevention Coalition conducted a community-based tuberculin screening and isoniazid preventive therapy project among high-risk inner-city residents of Atlanta, Georgia. We established screening centers in outpatient waiting areas of the public hospital serving inner-city residents, the city jail, clinics serving the homeless, and with outreach teams in neighborhoods frequented by drug users. All services were provided free. A total of 7,246 persons participated in tuberculin testing; 4,701 (65%) adhered with skin test reading, 809 (17%) had a positive test, 409 (50%) fit current guidelines for isoniazid preventive therapy, 84 (20%) we intended to treat completed therapy. The major limitations of this community-based tuberculin screening and preventive therapy project were the low proportion of infected individuals who were eligible for isoniazid preventive therapy and the poor adherence with a complete regimen among those we intended to treat. For community based programs to be efficacious, preventive therapy regimens that are of shorter duration and safe for older persons will need to be implemented. PMID- 9872854 TI - Role of lung perfusion scintigraphy in relation to chest computed tomography and pulmonary function in the evaluation of candidates for lung volume reduction surgery. AB - Lung perfusion scintigraphy is employed to evaluate patients with severe emphysema who are candidates for lung volume reduction surgery (LVRS). Our purpose was to investigate the role of scintigraphy in relation to chest computed tomography (CT) and lung function in this setting. Six observers blinded to clinical data retrospectively scored preoperative scintigrams of 70 patients undergoing bilateral video-assisted LVRS according to the distribution of lung perfusion as homogeneous, intermediately heterogeneous, or markedly heterogeneous. Heterogeneity of emphysema distribution was also assessed by chest CT. Dyspnea and pulmonary function were measured preoperatively and 3 mo postoperatively. In 42 patients with markedly heterogeneous, in 18 with intermediately heterogeneous, and in 10 with homogeneous perfusion, mean (+/- SE) FEV1 increased by 57 +/- 8% (p < 0.0001), 38 +/- 9% (p < 0.001), and 23 +/- 9% (p = NS) (p = NS for intergroup comparisons). In a multiple regression analysis, functional improvement after LVRS was more closely correlated with preoperative hyperinflation and the degree of emphysema heterogeneity estimated by chest CT than with the degree of perfusion heterogeneity assessed by scintigraphy. In 16 of 22 patients with homogeneous emphysema distribution in the chest CT scintigraphy revealed intermediately or markedly heterogeneous perfusion. We conclude that lung perfusion scintigraphy has a limited role in prediction of outcome, but it may help to identify target areas for resection in LVRS candidates with homogeneous CT morphology. PMID- 9872855 TI - Vital capacity reservoir and online measurement of childhood nitrosopnea are linearly related. Clinical implications. AB - Hypernitrosopnea, a robust marker for childhood asthma, is measured reproducibly in mixed vital capacity (VC) expirates. Recent guidelines for measurement of expired nitric oxide (NO) in adults have favored use of an online (OL), flow dependent technique. We compared VC and OL NO measurements in 14 asthmatic and 11 control children 5 through 18 yr of age. After spirometry, subjects breathed both into an open-ended reservoir (20 cm H2O resistance) and into a tedlar bag (VC maneuver). End-expiratory pressure > 5 cm H2O was continuously maintained during VC measurements, and the velum remained shut. Eight additional children (24% of total number of subjects) were unable reproducibly to perform the OL measurement at constant flow (six asthmatics; two control children). For subjects able to perform the OL technique, OL and VC NO measurements were linearly related (r2 = 0.88). In children, VC NO assays are reproducible, sensitive in identifying asthma, and portable. Additionally, we have shown that (1) not all children are able to perform OL measurements, and (2) VC measurements vary linearly with OL measurements. These findings suggest that there may not be compelling reason to favor OL over VC measurements for hypernitrosopnea in children with asthma. PMID- 9872856 TI - Medullary metastasis causing impairment of respiratory pressure output with intact respiratory rhythm. AB - We present an unusual case of weaning failure. A 67-yr-old man presented with confusion, hyponatremia, and hypercapnic respiratory failure that necessitated mechanical ventilation. CXR revealed a right hilar mass (non-small-cell carcinoma on biopsy). Level of consciousness improved with treatment of his hyponatremia. However, attempts at weaning were complicated by hypercapnia with no overt distress. Resistance and elastance were only slightly abnormal, excluding mechanics as a cause of respiratory failure. Maximal inspiratory pressure (MIP) and vital capacity (VC) were reduced at -15 cm H2O and 0.97 L, respectively. Limb muscle strength was well preserved, suggesting isolated respiratory muscle weakness. During a weaning trial respiratory rate increased from 7 to 40 breaths/min as PCO2 increased from 56 to 89 mm Hg, confirming an intact respiratory pacemaker and good response to CO2. However, spontaneous Pdi was only 1 to 2 cm H2O (< 20% of Pdimax) despite profound hypercapnia. The fact that the patient did not utilize a greater fraction of his pressure-generating capacity suggested preferential impairment of the automatic respiratory centers. MRI showed a large central metastatic lesion in the rostral medulla with only a thin rim of uninvolved tissue. This case illustrates the utility of relating the magnitude of spontaneous efforts to maximal voluntary efforts as a means of localizing the site of involvement in cases of respiratory muscle weakness. It also demonstrates that a large medullary mass lesion may selectively impair brainstem modulation of respiratory pressure output while sparing other medullary functions, and in particular the pacemaking function of the respiratory centers. PMID- 9872857 TI - Dyspnea. Mechanisms, assessment, and management: a consensus statement. American Thoracic Society. PMID- 9872858 TI - Being strategic about smoking. PMID- 9872859 TI - Low plasma vitamin D in Asian toddlers in Britain. PMID- 9872860 TI - Modernising mental health services. PMID- 9872861 TI - Opening up BMJ peer review. PMID- 9872862 TI - Treating thyrotoxicosis in pregnant or potentially pregnant women. PMID- 9872863 TI - Prisoner wants to donate his second kidney. PMID- 9872864 TI - Risks of surgery on unruptured brain aneurysms can outweigh benefits PMID- 9872865 TI - In brief PMID- 9872866 TI - Smoking ban improves health of bartenders PMID- 9872867 TI - WHO plans to halve child asthma deaths. PMID- 9872868 TI - UK treads softly on smoking. PMID- 9872869 TI - Shane Warne takes money to give up smoking. PMID- 9872871 TI - First doctor suspended under GMC performance procedures. PMID- 9872870 TI - Government plans shake up in NHS pay PMID- 9872873 TI - North Korean children ravaged by poor nutrition. PMID- 9872872 TI - Private company wins rights to Icelandic gene database. PMID- 9872874 TI - Death rates higher in Wales. PMID- 9872875 TI - Airline passenger dies after being sedated by doctor. PMID- 9872876 TI - Migraine and stroke in young women: case-control study. The World Health Organisation Collaborative Study of Cardiovascular Disease and Steroid Hormone Contraception. AB - OBJECTIVE: To investigate the association between migraine and ischaemic or haemorrhagic stroke in young women. DESIGN: Hospital based case-control study. SETTING: Five European centres participating in the World Health Organisation Collaborative Study of Cardiovascular Disease and Steroid Hormone Contraception. SUBJECTS: 291 women aged 20-44 years with ischaemic, haemorrhagic, or unclassified arterial stroke compared with 736 age and hospital matched controls. INTERVENTION: Questionnaire. MAIN OUTCOME MEASURE: Self reported history of headaches. RESULTS: Adjusted odds ratios associated with a personal history of migraine were 1.78 (95% confidence intervals, 1.14 to 2.77), 3.54 (1.30 to 9.61), and 1.10 (0.63 to 1.94) for all stroke, ischaemic stroke, and haemorrhagic stroke respectively. Odds ratios for ischaemic stroke were similar for classical migraine (with aura) (3.81, 1.26 to 11.5) and simple migraine (without aura) (2.97, 0.66 to 13.5). A family history of migraine, irrespective of personal history, was also associated with increased odds ratios, not only for ischaemic stroke but also haemorrhagic stroke. In migrainous women, coexistent use of oral contraceptives or a history of high blood pressure or smoking had greater than multiplicative effects on the odds ratios for ischaemic stroke associated with migraine alone. Change in the frequency or type of migraine on using oral contraceptives did not predict subsequent stroke. Between 20% and 40% of strokes in women with migraine seemed to develop directly from a migraine attack. CONCLUSIONS: Migraine in women of childbearing age significantly increases the risk of ischaemic but not haemorrhagic stroke. The coexistence of oral contraceptive use, high blood pressure, or smoking seems to exert a greater than multiplicative effect on the risk of ischaemic stroke associated with migraine. PMID- 9872877 TI - Social class in childhood and general health in adulthood: questionnaire study of contribution of psychological attributes. AB - OBJECTIVE: To determine the contribution of psychological attributes (personality characteristics and coping styles) to the association between social class in childhood and adult health among men and women. DESIGN: Partly retrospective, partly cross sectional study conducted in the framework of the Dutch GLOBE study. SUBJECTS: Sample of general population from south east Netherlands consisting of 2174 men and women aged 25-74 years. Baseline self reported data from 1991 provided information on childhood and adult social class, psychological attributes, and general health. MAIN OUTCOME MEASURE: Self rated poor health. RESULTS: Independent of adult social class, low childhood social class was related to self rated poor health (odds ratio 1.67 (95% confidence interval 1.02 to 2.75) for subjects whose fathers were unskilled manual workers versus subjects whose fathers were higher grade professionals). Subjects whose fathers were manual workers generally had more unfavourable personality profiles and more negative coping styles. External locus of control, neuroticism, and the absence of active problem focused coping explained about half of the association between childhood social class and self rated poor health. The findings were independent of adult social class and height. CONCLUSIONS: A higher prevalence of negative personality profiles and adverse coping styles in subjects who grew up in lower social classes explains part of the association between social class in childhood and adult health. This finding underlines the importance of psychological mechanisms in the examination of the negative effects of adverse socioeconomic conditions in childhood. PMID- 9872878 TI - Effect of open peer review on quality of reviews and on reviewers' recommendations: a randomised trial. AB - OBJECTIVES: To examine the effect on peer review of asking reviewers to have their identity revealed to the authors of the paper. DESIGN: Randomised trial. Consecutive eligible papers were sent to two reviewers who were randomised to have their identity revealed to the authors or to remain anonymous. Editors and authors were blind to the intervention. MAIN OUTCOME MEASURES: The quality of the reviews was independently rated by two editors and the corresponding author using a validated instrument. Additional outcomes were the time taken to complete the review and the recommendation regarding publication. A questionnaire survey was undertaken of the authors of a cohort of manuscripts submitted for publication to find out their views on open peer review. RESULTS: Two editors' assessments were obtained for 113 out of 125 manuscripts, and the corresponding author's assessment was obtained for 105. Reviewers randomised to be asked to be identified were 12% (95% confidence interval 0.2% to 24%) more likely to decline to review than reviewers randomised to remain anonymous (35% v 23%). There was no significant difference in quality (scored on a scale of 1 to 5) between anonymous reviewers (3.06 (SD 0.72)) and identified reviewers (3.09 (0.68)) (P=0.68, 95% confidence interval for difference - 0.19 to 0.12), and no significant difference in the recommendation regarding publication or time taken to review the paper. The editors' quality score for reviews (3.05 (SD 0.70)) was significantly higher than that of authors (2.90 (0.87)) (P<0.005, 95%confidence interval for difference - 0.26 to - 0.03). Most authors were in favour of open peer review. CONCLUSIONS: Asking reviewers to consent to being identified to the author had no important effect on the quality of the review, the recommendation regarding publication, or the time taken to review, but it significantly increased the likelihood of reviewers declining to review. PMID- 9872879 TI - Vitamin D concentrations in Asian children aged 2 years living in England: population survey. PMID- 9872880 TI - Incidence of trauma related stress fractures and shin splints in male and female army recruits: retrospective case study. PMID- 9872882 TI - The health administration of london PMID- 9872881 TI - Gide's solution PMID- 9872883 TI - Qualitative study of decisions about infant feeding among women in east end of London. AB - OBJECTIVE: To improve understanding of how first time mothers who belong to a socioeconomic group with particularly low rates of breast feeding decide whether or not to initiate breast feeding. DESIGN: Qualitative semistructured interviews early in pregnancy and 6-10 weeks after birth. SETTING: Women's homes in east end of London. SUBJECTS: 21 white, low income women expecting their first baby were interviewed mostly at home, often with their partner or a relative. Two focus groups were conducted. RESULTS: Women who had regularly seen a relative or friend successfully breast feed and described this experience positively were more confident about and committed to breast feeding. They were also more likely to succeed. Exposure to breast feeding, however, could be either a positive or a negative influence on the decision to breast feed, depending on the context. Women who had seen breast feeding only by a stranger often described this as a negative influence, particularly if other people were present. All women knew that breast feeding has health benefits. Ownership of this knowledge, however, varied according to the woman's experience of seeing breast feeding. CONCLUSIONS: The decision to initiate breast feeding is influenced more by embodied knowledge gained from seeing breast feeding than by theoretical knowledge about its benefits. Breast feeding involves performing a practical skill, often with others present. The knowledge, confidence, and commitment necessary to breast feed may be more effectively gained through antenatal apprenticeship to a breastfeeding mother than from advice given in consultations or from books. PMID- 9872884 TI - Recent advances: neurosurgery. PMID- 9872886 TI - The knock on the door PMID- 9872885 TI - Lesson of the week: florid rickets associated with prolonged breast feeding without vitamin D supplementation. PMID- 9872887 TI - The knock on the door PMID- 9872889 TI - Special correspondence: paris PMID- 9872888 TI - ABC of sexual health: female sexual problems I: loss of desire-what about the fun? PMID- 9872890 TI - Evidence on peer review-scientific quality control or smokescreen? PMID- 9872891 TI - Rethinking transparency and accountability in medicines regulation in the United Kingdom. PMID- 9872893 TI - A military doctor PMID- 9872892 TI - Narrative based medicine: why study narrative? PMID- 9872894 TI - Useful remedy PMID- 9872895 TI - Suboptimal ward care of critically ill patients. Suboptimal care should have been defined. PMID- 9872896 TI - Complaints of pain after use of handcuffs should not be dismissed. PMID- 9872897 TI - Electronic preprints can be categorised. PMID- 9872898 TI - Meta-analyses of observational data should be done with due care. PMID- 9872899 TI - Infantile spasms and vigabatrin. Study will compare effects of drugs. PMID- 9872900 TI - Sex education should begin in primary school. PMID- 9872901 TI - Improved command of English may explain why non-English speaking countries get more published. PMID- 9872902 TI - Is it time for an Illich Collaboration to make available information on the harms of medical care? PMID- 9872903 TI - Clinical outcome in relation to care in centres specialising in cystic fibrosis. Cross infection with Pseudomonas aeruginosa is unusual. PMID- 9872904 TI - Treatment of gastro-oesophageal reflux disease in adults. Efficacy of surgery needs to be compared with that of proton pump inhibitors. PMID- 9872905 TI - Communicating with patients. Specialist training should include communication skills. PMID- 9872906 TI - Author was never appointed to bristol inquiry PMID- 9872907 TI - Nilima choudhury, nee kundu PMID- 9872908 TI - BMA calls for guidelines on new and expensive drugs PMID- 9872909 TI - Desperately seeking solutions: rationing health care PMID- 9872910 TI - Gout: the patrician malady PMID- 9872911 TI - Step by step guide to epidemiological health needs assessment for ethnic minority groups PMID- 9872912 TI - Minor surgery and skin lesions: diagnosis and management on CD-ROM PMID- 9872913 TI - Shock to stop? PMID- 9872914 TI - Website of the week PMID- 9872915 TI - My route to a steroid psychosis PMID- 9872917 TI - Ethics and narrative PMID- 9872916 TI - Bookcase PMID- 9872919 TI - Psychological attributes may explain how social origins affect adult health PMID- 9872918 TI - Migraine increases the risk of ischaemic but not haemorrhagic stroke in young women PMID- 9872920 TI - Open peer review is feasible and does not reduce quality of reviews PMID- 9872921 TI - Breast feeding is best learnt by example PMID- 9872922 TI - Asian children in the UK may need vitamin D PMID- 9872923 TI - Medicines regulation should be more transparent PMID- 9872924 TI - Regulation of G-protein-coupled receptor kinase subtypes by calcium sensor proteins. AB - The process of G-protein-coupled receptor (GPCR) homologous desensitization is intrinsically related to the function of a class of S/T kinases named G-protein coupled receptor kinases (GRK). GRK family is so far composed of six cloned members, named GRK1 to 6, which are classified into three subfamilies: GRK1 is alone in the first (rhodopsin kinase subfamily), GRK2 and 3 form the second [beta adrenergic receptor kinase (betaARK) subfamily], and GRK4, 5, and 6 constitute the third (GRK4 subfamily). Recent studies from different laboratories have demonstrated that different calcium sensor proteins (CSP) can selectively regulate the activity of GRK subtypes. In the presence of calcium, rhodopsin kinase (GRK1) is inhibited by the photoreceptor-specific CSP recoverin through direct binding. Several other recoverin homologues (including NCS 1, VILIP 1, and hippocalcin) are also able to inhibit GRK1 in a calcium-dependent manner. The ubiquitous calcium binding protein calmodulin (CaM) can inhibit GRK5 with a high affinity (IC50=40-50 nM). A direct interaction between GRK5 and Ca2+/CaM was documented and this binding did not influence the catalytic activity of the kinase, but rather reduced GRK5 binding to the membrane. These studies suggest that CSP act as functional analogs in mediating the regulation of different GRK subtypes by Ca2+. This mechanism, however, is highly selective with respect to the GRK subtypes: GRK1, but not GRK2 and GRK5, is regulated by recoverin and other NCS, but GRK4, 5, and 6, which belong to the GRK4 subfamily are potently inhibited by CaM, which has little or no effect on members of other GRK subfamilies. Calcium-dependent inhibition of rhodopsin kinase by recoverin represents one of the mechanisms that control adaptation to light. For the other GPCR, CSP-GRK interaction provides a feedback mechanism that can modulate homologous desensitization of these receptors. PMID- 9872925 TI - Vascular endothelial growth factor (VEGF) and its receptors. AB - Vascular endothelial growth factor (VEGF) is a highly specific mitogen for vascular endothelial cells. Five VEGF isoforms are generated as a result of alternative splicing from a single VEGF gene. These isoforms differ in their molecular mass and in biological properties such as their ability to bind to cell surface heparan-sulfate proteoglycans. The expression of VEGF is potentiated in response to hypoxia, by activated oncogenes, and by a variety of cytokines. VEGF induces endothelial cell proliferation, promotes cell migration, and inhibits apoptosis. In vivo VEGF induces angiogenesis as well as permeabilization of blood vessels, and plays a central role in the regulation of vasculogenesis. Deregulated VEGF expression contributes to the development of solid tumors by promoting tumor angiogenesis and to the etiology of several additional diseases that are characterized by abnormal angiogenesis. Consequently, inhibition of VEGF signaling abrogates the development of a wide variety of tumors. The various VEGF forms bind to two tyrosine-kinase receptors, VEGFR-1 (flt-1) and VEGFR-2 (KDR/flk 1), which are expressed almost exclusively in endothelial cells. Endothelial cells express in addition the neuropilin-1 and neuropilin-2 coreceptors, which bind selectively to the 165 amino acid form of VEGF (VEGF165). This review focuses on recent developments that have widened considerably our understanding of the mechanisms that control VEGF production and VEGF signal transduction and on recent studies that have shed light on the mechanisms by which VEGF regulates angiogenesis. PMID- 9872926 TI - Contributions of polyol pathway to oxidative stress in diabetic cataract. AB - There is strong evidence to show that diabetes is associated with increased oxidative stress. However, the source of this oxidative stress remains unclear. Using transgenic mice that overexpress aldose reductase (AR) in their lenses, we found that the flux of glucose through the polyol pathway is the major cause of hyperglycemic oxidative stress in this tissue. The substantial decrease in the level of reduced glutathione (GSH) with concomitant rise in the level of lipid peroxidation product malondialdehyde (MDA) in the lens of transgenic mice, but not in the nontransgenic mice, suggests that glucose autoxidation and nonenzymatic glycation do not contribute significantly to oxidative stress in diabetic lenses. AR reduction of glucose to sorbitol probably contributes to oxidative stress by depleting its cofactor NADPH, which is also required for the regeneration of GSH. Sorbitol dehydrogenase, the second enzyme in the polyol pathway that converts sorbitol to fructose, also contributes to oxidative stress, most likely because depletion of its cofactor NAD+ leads to more glucose being channeled through the polyol pathway. Despite a more than 100% increase of MDA, oxidative stress plays only a minor role in the development of cataract in this acute diabetic cataract model. However, chronic oxidative stress generated by the polyol pathway is likely to be an important contributing factor in the slow developing diabetic cataract as well as in the development of other diabetic complications.--Lee, A. Y. W., Chung, S. S. M. Contributions of polyol pathway to oxidative stress in diabetic cataract. FASEB J. 13, 23-30 (1999) PMID- 9872927 TI - A Sos-derived peptidimer blocks the Ras signaling pathway by binding both Grb2 SH3 domains and displays antiproliferative activity. AB - With the aim of interrupting the growth factor-stimulated Ras signaling pathway at the level of the Grb2-Sos interaction, a peptidimer, made of two identical proline-rich sequences from Sos linked by a lysine spacer, was designed using structural data from Grb2 and a proline-rich peptide complexed with its SH3 domains. The peptidimer affinity for Grb2 is 40 nM whereas that of the monomer is 16 microM, supporting the dual recognition of both Grb2 SH3 domains by the dimer. At 50 nM, the peptidimer blocks selectively Grb2-Sos complexation in ER 22 (CCL 39 fibroblasts overexpressing epidermal growth factor receptor) cellular extracts. The peptidimer specifically recognizes Grb2 and does not interact with PI3K or Nck, two SH3 domain-containing adaptors. The peptidimer was modified to enter cells by coupling to a fragment of Antennapedia homeodomain. At 10 microM, the conjugate inhibits the Grb2-Sos interaction (100%) and MAP kinase (ERK1 and ERK2) phosphorylation (60%) without modifying cellular growth of ER 22 cells. At the same concentration, the conjugate also inhibits both MAP kinase activation induced by nerve growth factor or epidermal growth factor in PC12 cells, and differentiation triggered by nerve growth factor. Finally, when tested for its antiproliferative activity, the conjugate was an efficient inhibitor of the colony formation of transformed NIH3T3/HER2 cells grown in soft agar, with an IC50 of around 1 microM. Thus, the designed peptidimers appear to be interesting leads to investigate signaling and intracellular processes and for designing selective inhibitors of tumorigenic Ras-dependent processes. PMID- 9872928 TI - Toward the experimental codon reassignment in vivo: protein building with an expanded amino acid repertoire. AB - The high precision and fidelity of the genetic message transmission are ensured by numerous proofreading steps, from DNA replication and transcription to protein translation. The key event for translational fidelity is the proper codon assignment for 20 canonical amino acids. An experimental codon reassignment is possible for noncanonical amino acids in vivo using artificially constructed expression hosts under efficient selective pressure. However, such amino acids may interfere with the cellular metabolism and thus do not belong to the 'first' or 'restricted' part of the universal code, but rather to a second or 'relaxed' part, which is limited mainly by the downstream proofreading in the natural translational machinery. Correspondingly, not all possible alpha-amino acids can be introduced into proteins. The aim of this study is to discuss biological and evolutionary constraints on possible candidates for this second coding level of the universal code. Engineering of such a 'second' code is expected to have great academic as well as practical impact, ranging from protein folding studies to biomedicine. PMID- 9872929 TI - Identification of laminin alpha1 and beta1 chain peptides active for endothelial cell adhesion, tube formation, and aortic sprouting. AB - Laminin-1 is a basement membrane glycoprotein that promotes several biological activities including cell attachment, tumor metastasis, and angiogenesis. Angiogenesis plays an important role in tissue formation, reproduction, wound healing, and several pathological conditions. In this study, we screened 405 synthetic peptides from the alpha1 and beta1 chains to identify potential sites on laminin-1 active with endothelial cells. Peptides were initially screened by testing both endothelial cell adhesion to peptide-coated wells and tube formation on Matrigel in the presence of soluble peptide. Twenty active peptides were identified in these screens. A secondary screen using the rat aortic ring sprouting assay identified 13 of the 20 peptides that stimulated endothelial sprouting. Several of these active peptides were also found to stimulate human umbilical vein endothelial cell migration in Boyden chamber assays. Differences in the amount of peptide needed for the response and in the resultant morphologies/responses were observed between the peptides in all of the assays. Our results suggest that several active domains on laminin-1 may play important roles in stimulating different steps in angiogenesis. PMID- 9872930 TI - TNFalpha plus IFNgamma induce the production of Alzheimer beta-amyloid peptides and decrease the secretion of APPs. AB - The appearance of inflammatory markers associated with amyloid plaques indicates a state of chronic inflammation in Alzheimer's disease (AD). Multiple epidemiological studies also suggest that patients taking anti-inflammatory drugs have a decreased risk of developing AD. Here we present evidence that inflammatory cytokines can alter the metabolism of the beta-amyloid precursor protein (betaAPP). We show that the combination of tumor necrosis factor alpha and interferon gamma triggers the production of beta-amyloid peptides and inhibits the secretion of soluble APPs by human neuronal and extraneuronal cells. The results demonstrate a new mechanism by which inflammatory components can exacerbate the fundamental pathology in AD. PMID- 9872931 TI - Hydrogen peroxide produced during gamma-glutamyl transpeptidase activity is involved in prevention of apoptosis and maintainance of proliferation in U937 cells. AB - It has been reported in several cell lines that exposure to low levels of reactive oxygen species can exert a stimulatory effect on their proliferation. We have previously shown that mild oxidative conditions can also counteract apoptotic stimuli. A constitutive cellular production of low levels of superoxide and hydrogen peroxide originates from various sources; among these, gamma glutamyl transpeptidase (GGT), the plasma membrane-bound activity in charge of metabolizing extracellular reduced glutathione, has recently been included. Since the inhibition of GGT is a sufficient stimulus for the induction of apoptosis in selected cell lines, we investigated whether this effect might result from the suppression of the mentioned GGT-dependent prooxidant reactions, on the theory that the latter may represent a basal antiapoptotic and proliferative signal for the cell. Experiments showed that: 1) GGT activity in U937 monoblastoid cells is associated with the production of low levels of hydrogen peroxide, and two independent GGT inhibitors cause a dose-dependent decrease of such GGT-dependent production of H2O2; 2) GGT inhibition with acivicin results in cell growth arrest, and induces cell death and DNA fragmentation with the ladder appearance of apoptosis; 3) treatment of cells with catalase--and even more with Trolox C- is able to decrease their proliferative rate; 4) GGT inhibition (with suppression of H2O2 production) results in a down-regulation of poly(ADP-ribose) polimerase (PARP) activity, which precedes the proteolytic cleavage of PARP molecule, such as that typically induced by caspases. The reported data suggest that the low H2O2 levels originating as a by-product during GGT activity are able to act as sort of a 'life signal' in U937 cells, insofar as they can maintain cell proliferation and protect against apoptosis, possibly through an up-regulation of PARP activity. PMID- 9872932 TI - General applicability of chicken egg yolk antibodies: the performance of IgY immunoglobulins raised against the hypoxia-inducible factor 1alpha. AB - Avian embryos and neonates acquire passive immunity by transferring maternal immunoglobulins from serum to egg yolk. Despite being a convenient source of antibodies, egg yolk immunoglobulins (IgY) from immunized hens have so far received scant attention in research. Here we report the generation and rapid isolation of IgY from the egg yolk of hens immunized against the alpha subunit of the human hypoxia-inducible factor 1 (HIF-1alpha). Anti-HIF-1alpha IgY antibodies were affinity purified and tested for their performance in various applications. Abundant HIF-1alpha protein was detected by Western blot analysis in nuclear extracts derived from hypoxic cells of human, mouse, monkey, swine, and dog origin whereas in hypoxic quail and frog cells, the HIF-1alpha signal was weak or absent, respectively. In electrophoretic mobility shift assays, affinity-purified IgY antibody was shown to recognize the native HIF-1 (but not the related HIF-2) complex that specifically binds an oligonucleotide containing the HIF-1 DNA binding site. Furthermore, IgY antibody immunoprecipitated HIF-1alpha from hypoxic cell extracts. Immunofluorescence experiments using IgY antibody allowed the detection of HIF-1alpha in the nucleus of hypoxic COS-7 cells. For comparison, the application of a mouse monoclonal antibody raised against the identical HIF-1alpha fragment was more restricted. Because chicken housing is inexpensive, egg collection is noninvasive, isolation and affinity purification of IgY antibodies are fast and simple, and the applicability of IgY is widespread, immunization of hens represents an excellent alternative for the generation of polyclonal antibodies. PMID- 9872933 TI - Reduced levels of poly(ADP-ribosyl)ation result in chromatin compaction and hypermethylation as shown by cell-by-cell computer-assisted quantitative analysis. AB - The unmethylated status of the CpG islands is important for gene expression of correlated housekeeping genes since it is well known that their methylation inhibits transcription process. An interesting question that has been discussed but not solved is how the CpG islands maintain their characteristic unmethylated status even though they are rich in CpG dinucleotides. Our previous in vitro and in vivo research has shown that poly(ADP-ribosyl)ation is involved in protecting CpG dinucleotides from full methylation in genomic DNA and that a block of poly(ADP-ribosyl)ation is also involved in modifying the methylation pattern in the promoter region of Htf9 housekeeping gene. In this study we locked for cytological evidence that in the absence of an active poly(ADP-ribosyl)ation the DNA methylation pattern in L929 and NIH/3T3 mouse fibroblast cell lines is altered. For this purpose, differences in the methylation levels of interphase nuclei from control and treated cultures of two murine cell lines preincubated with 2 mM 3-aminobenzamide, an inhibitor of poly(ADP-ribosyl)ation, were measured in individual cells after indirect immunolabeling with anti-5MeC antibodies. The quantitative analysis allowed us to demonstrate that blocking of the poly(ADP ribosyl)ation results in a higher number, size, and density of antibody binding regions in treated cells when compared to the controls. Analogously, sequential Giemsa staining and indirect immunolabeling of the same slides showed the heterochromatic regions colocalized with the extended methyl-rich domains. PMID- 9872934 TI - Magnetic fields increase cell survival by inhibiting apoptosis via modulation of Ca2+ influx. AB - Static magnetic fields with intensities starting from 6 gauss (6x10(-4) tesla, T) were found to decrease in an intensity-dependent fashion, reaching a plateau at 6 x 10(-3) T, the extent of cell death by apoptosis induced by several agents in different human cell systems. This is not due to a change in the mode of cell death (i.e., to necrosis) or to a delay of the process itself; rather, the presence of magnetic fields allows the indefinite survival and replication of the cells hit by apoptogenic agents. The protective effect was found to be mediated by the ability of the fields to enhance Ca2+ influx from the extracellular medium; accordingly, it was limited to those cell systems where Ca2+ influx was shown to have an antiapoptotic effect. Magnetic fields thus might interfere with human health by altering/restoring the equilibrium between cell death and proliferation; indeed, the rescue of damaged cells may be the mechanism explaining why magnetic fields that are not mutagenic per se are often able to increase mutation and tumor frequencies. PMID- 9872935 TI - Acute hyperglycemia regulates transcription and posttranscriptional stability of PKCbetaII mRNA in vascular smooth muscle cells. AB - Acute hyperglycemia may contribute to the progression of atherosclerosis by regulating protein kinase C (PKC) isozymes and by accelerating vascular smooth muscle cell (VSMC) proliferation. We investigated acute glucose regulation of PKCbeta gene expression in A10 cells, a rat aortic smooth muscle cell line. Western blot analysis showed that PKCbetaII protein levels decreased with high glucose (25 mM) compared to normal glucose (5.5 mM), whereas PKCbetaI levels were unaltered. PKCbeta mRNA levels were depleted by 60-75% in hyperglycemic conditions. To elucidate whether high glucose regulated PKCbeta expression via the common promoter for PKCbetaI and PKCbetaII, deletion constructs of the PKCbeta promoter ligated to CAT as reporter gene were transfected into A10 cells. Construct D (-411 to +179CAT) showed quenching in high glucose (25 mM) and suggested the involvement of a carbohydrate response element in the 5' promoter region of the PKCbeta gene. In actinomycin D-treated A10 cells, a 60% decrease in PKCbeta mRNA with high glucose treatment indicated that posttranscriptional destabilization by glucose was also occurring. We have demonstrated that glucose induced posttranscriptional destabilization of PKCbetaII message is mediated via a nuclease activity present in the cytosol. The specificity of glucose to posttranscriptionally destabilize PKCbetaII mRNA, but not the PKCbetaI mRNA, was confirmed in both A10 cells and primary cultures from human aorta. PMID- 9872936 TI - Plasticity of hippocampal corticosteroid receptors during aging in the rat. AB - Aging is commonly associated with dysregulation of the hypothalamo-pituitary adrenal axis and cognitive impairment. On the basis of suggestions that these disruptions ensue from changes in the hippocampal complement of corticosteroid (mineralocorticoid and glucocorticoid) receptors (MR and GR), we examined the availability of hippocampal MR and GR by measuring the in vivo uptake of 3H aldosterone and 3H-dexamethasone (selective MR and GR agonists, respectively); MR and GR mRNA levels were also measured. We observed age-related declines in both the synthesis of MR and GR and the uptake of their respective ligands. Whereas MR mRNA levels and ligand uptake declined in parallel, GR binding declined more steeply than GR mRNA. This latter result, together with our finding that aged rats show impaired corticosteroid receptor mRNA and protein up-regulation after corticosteroid withdrawal, indicates decreased transcription of MR and GR genes and posttranslational modification of GR mRNA during aging. Given that corticosteroids can influence MR and GR synthesis and binding, and based on the finding that aged subjects show reduced basal secretion of corticosterone, we propose that this relative hypocorticalism may be responsible for the changes observed in MR and GR activity, which then leads to disturbances in neuroendocrine regulation and cognitive function in aged subjects. PMID- 9872937 TI - Functional testosterone receptors in plasma membranes of T cells. AB - T cells are considered to be unresponsive to testosterone due to the absence of androgen receptors (AR). Here, we demonstrate the testosterone responsiveness of murine splenic T cells in vitro as well as the presence of unconventional cell surface receptors for testosterone and classical intracellular AR. Binding sites for testosterone on the surface of both CD4(+) and CD8(+) subsets of T cells are directly revealed with the impeded ligand testosterone-BSA-FITC by confocal laser scanning microscopy (CLSM) and flow cytometry, respectively. Binding of the plasma membrane impermeable testosterone-BSA conjugate induces a rapid rise (<5 s) in [Ca2+]i of Fura-2-loaded T cells. This rise reflects influx of extracellular Ca2+ through non-voltage-gated and Ni2+-blockable Ca2+ channels of the plasma membrane. The testosterone-BSA-induced Ca2+ import is not affected by cyproterone, a blocker of the AR. In addition, AR are not detectable on the surface of intact T cells when using anti-AR antibodies directed against the amino and carboxy terminus of the AR, although T cells contain AR, as revealed by reverse transcription-polymerase chain reactions and Western blotting. AR can be visualized with the anti-AR antibodies in the cytoplasm of permeabilized T cells by using CLSM, though AR are not detectable in cytosol fractions when using the charcoal binding assay with 3H-R1881 as ligand. Cytoplasmic AR do not translocate to the nucleus of T cells in the presence of testosterone, in contrast to cytoplasmic AR in human cancer LNCaP cells. These findings suggest that the classical AR present in splenic T cells are not active in the genomic pathway. By contrast, the cell surface receptors for testosterone are in a functionally active state, enabling T cells a nongenomic response to testosterone. PMID- 9872938 TI - Choline deficiency-induced apoptosis in PC12 cells is associated with diminished membrane phosphatidylcholine and sphingomyelin, accumulation of ceramide and diacylglycerol, and activation of a caspase. AB - It is not well appreciated that nutritional status can modulate apoptosis, a process that eliminates unwanted or damaged cells. Choline is an essential nutrient, and its absence induces apoptosis. When PC12 cells were cultivated in a choline-free medium, apoptosis was induced (27.4% of cells apoptotic at 72 h as compared to 4.4% in control medium). In choline-free medium at 72 h, there was a 49% decrease in phosphatidylcholine concentration (P<0.01) and a 34% decrease in sphingomyelin concentration (P<0.01); however, there was no change in phosphatidylethanolamine concentration. Before detecting increased apoptosis in choline-deficient cells, we measured a significant increase in ceramide (218% control) and diacyglycerol (155% control) concentrations. The addition of a cell permeable ceramide to cells in control medium induced apoptosis; however, adding a cell-permeable diacyglycerol did not induce apoptosis. Caspase is a common mediator of apoptosis, and choline deficiency-induced apoptosis was prevented completely by replacing choline or adding a caspase inhibitor into the medium within 48 h of initial choline deprivation. In those cells rescued by replacing choline at 36 h, the concentrations of phosphatidylcholine, sphingomyelin, ceramide, and diacyglycerol returned to levels of control cells. In those cells rescued by adding a caspase inhibitor at 36 h, the concentrations of sphingomyelin and ceramide returned to control levels, but the concentrations of phosphatidylcholine and diacyglycerol did not return to normal. We propose that availability of dietary factors (choline in this model) can modulate apoptosis. Mechanisms that we identify using this model may help us to explain why dietary choline influences brain development. PMID- 9872939 TI - Inverse regulation of lipid-peroxidizing and hydroperoxyl lipid-reducing enzymes by interleukins 4 and 13. AB - 12/15-lipoxygenases and phospholipid hydroperoxide glutathione peroxidases are opposite enzymes balancing the intracellular concentration of hydroperoxy lipids. We studied the regulation of both enzymes by interleukins 4 and 13 and found an inverse response. When human lung carcinoma cells A549 were cultured in vitro in the presence of these cytokines, an up-regulation of the 12/15-lipoxygenase and a down-regulation of the phospholipid hydroperoxide glutathione peroxidase were observed. A similar inverse regulation was found in human peripheral monocytes. Interleukin 4-treated A549 cells exhibited an impaired capability of reducing exogenous hydroperoxyl lipids and their levels of endogenous lipid hydroperoxides were markedly increased. To find out whether these regulatory processes also occur in vivo, arachidonic acid oxygenase and phospholipid hydroperoxide glutathione peroxidase activity was assayed in various tissues of transgenic mice that systemically overexpress interleukin 4. In lung, spleen, kidney, and heart, an increased arachidonic acid oxygenase activity was detected when transgenic mice were compared with inbred controls. The phospholipid hydroperoxide glutathione peroxidase activity was impaired in lung, liver, and spleen of the transgenic animals. These data indicate that lipid-peroxidizing and lipid peroxide-reducing enzymes are inversely regulated in various mammalian cells. Up regulation of the 12/15-lipoxygenase and simultaneous down-regulation of the phospholipid hydroperoxide glutathione peroxidase may lead to an increased oxidizing potential, which is reflected by an augmented intracellular peroxide tone. PMID- 9872940 TI - Cold-induced apoptosis in cultured hepatocytes and liver endothelial cells: mediation by reactive oxygen species. AB - When cultured hepatocytes were incubated in cell culture medium at 4 degreesC for up to 30 h and then returned to 37 degreesC, blebbing of the plasma membrane, cell detachment, chromatin condensation and margination, enhanced nuclear stainability with Hoechst 33342, ruffling of the nuclear membrane, and DNA fragmentation occurred. Similar to hepatocytes, cultured liver endothelial cells exhibited blebbing, chromatin condensation and margination, marked nuclear condensation, and increased stainability with Hoechst 33342 when exposed to hypothermia/rewarming. In both cell types, the occurrence and extent of these alterations were dependent on the duration of the cold incubation period. This cold-induced apoptosis was inhibited by hypoxia, by an array of free radical scavengers/antioxidants, and by iron chelators. However, the extent of the protection by the different antioxidants was different in the two cell types: iron chelators provided complete protection in liver endothelial cells but only partial protection in hepatocytes, whereas lipophilic antioxidants such as alpha tocopherol provided complete protection in both cell types. During cold incubation, and especially during rewarming, lipid peroxidation occurred. These results suggest that the formation of reactive oxygen species (ROS) is a key mediator of cold-induced apoptosis, with ROS formation being completely iron mediated in liver endothelial cells and partially iron-mediated in hepatocytes. PMID- 9872941 TI - Dysfunctional glucocorticoid receptor with a single point mutation ablates the CCAAT/enhancer binding protein-dependent growth suppression response in a steroid resistant rat hepatoma cell variant. AB - We used glucocorticoid-resistant and -sensitive hepatoma cell variants to characterize the mechanism of hepatoma cell resistance to the growth inhibitory effects of glucocorticoids. BDS1 hepatoma cells express transcriptionally active glucocorticoid receptors and undergo a stringent G1 cell cycle arrest in response to glucocorticoids that is dependent on the induced expression of the CCAAT/enhancer binding protein alpha (C/EBPalpha) transcription factor. In contrast, EDR1 hepatoma cells, which express normal levels of glucocorticoid receptors, fail to growth arrest or express C/EBPalpha when treated with glucocorticoids. Ectopic expression of wild-type rat glucocorticoid receptors into EDR1 cells restored the growth suppression response, suggesting a defect in the EDR1 receptor. DNA sequence analysis revealed a single point mutation causing a cysteine-to-tyrosine substitution at amino acid position 457 (C457Y-GR) in the zinc finger region of the glucocorticoid receptor that mediates both receptor-DNA and receptor-protein interactions. Glucocorticoid activation of the alpha1-acid glycoprotein (AGP) promoter, a liver acute-phase response gene, requires receptor DNA binding as well as an interaction with C/EBPalpha. In contrast to the wild type glucocorticoid receptor, ectopic expression of C/EBPalpha in EDR1 cells, or coexpression of C/EBPalpha along with the C457Y-GR into receptor-deficient EDR3 cells was required to partially restore glucocorticoid responsiveness of the AGP promoter by the EDR1 glucocorticoid receptor. Constitutive expression of the wild type glucocorticoid receptor, but not the C457Y-GR mutant, was sufficient to restore the glucocorticoid growth suppression response to receptor-deficient EDR3 cells. Thus, we have identified a glucocorticoid-resistant hepatoma cell variant with a single point mutation in the zinc finger region of the glucocorticoid receptor gene that ablates the glucocorticoid growth suppression response and attenuates transcriptional activation of the AGP promoter. PMID- 9872942 TI - VEGI, a novel cytokine of the tumor necrosis factor family, is an angiogenesis inhibitor that suppresses the growth of colon carcinomas in vivo. AB - A novel member of the tumor necrosis factor (TNF) family has been identified from the human umbilical vein endothelial cell cDNA library, named vascular endothelial growth inhibitor (VEGI). The VEGI gene was mapped to human chromosome 9q32. The cDNA for VEGI encodes a protein of 174 amino acid residues with the characteristics of a type II transmembrane protein. Its amino acid sequence is 20 30% identical to other members of the TNF family. Unlike other members of the TNF family, VEGI is expressed predominantly in endothelial cells. Local production of a secreted form of VEGI via gene transfer caused complete suppression of the growth of MC-38 murine colon cancers in syngeneic C57BL/6 mice. Histological examination showed marked reduction of vascularization in MC-38 tumors that expressed soluble but not membrane-bound VEGI or were transfected with control vector. The conditioned media from soluble VEGI-expressing cells showed marked inhibitory effect on in vitro proliferation of adult bovine aortic endothelial cells. Our data suggest that VEGI is a novel angiogenesis inhibitor of the TNF family and functions in part by directly inhibiting endothelial cell proliferation. The results further suggest that VEGI maybe highly valuable toward angiogenesis-based cancer therapy. PMID- 9872943 TI - Modulation of mitochondrial respiration by nitric oxide: investigation by single cell fluorescence microscopy. AB - With the electro-driven import of rhodamine 123, we used single cell fluorescence microscopy to single out the contribution of nitric oxide (NO) in controlling mitochondrial membrane potential expressed by (stationary growing) rhabdomyosarcoma and neuroblastoma cells in culture. The experimental design and the computer-aided image analysis detected and quantitated variations of fluorescence signals specific to mitochondria. We observed that 1) the two cell lines display changes of fluorescence dependent on mitochondrial energization states; 2) mitochondrial fluorescence decreases after exposure of the cells to a NO releaser; 4) the different fluorescence intensity measured under stationary growing conditions, or after activation and inhibition of constitutive NO synthase, is consistent with a steady-state production of NO. Direct comparison of single cell fluorescence with bulk cytofluorimetry proved that the results obtained by the latter method may be misleading because of the intrinsic-to measure lack of information about distribution of fluorescence within different cell compartments. The kinetic parameters describing the reactions between cytochrome oxidase, NO, and O2 may account for the puzzling (20-fold) increase of the KM for O2 reported for cells and tissues as compared to purified cytochrome c oxidase, allowing an estimate of in vivo NO flux. PMID- 9872944 TI - Chlamydomonas: the cell and its genomes. PMID- 9872946 TI - A region of the Sir1 protein dedicated to recognition of a silencer and required for interaction with the Orc1 protein in saccharomyces cerevisiae. AB - Silencing of the cryptic mating-type loci HMR and HML requires the recognition of DNA sequence elements called silencers by the Sir1p, one of four proteins dedicated to the assembly of silenced chromatin in Saccharomyces cerevisiae. The Sir1p is thought to recognize silencers indirectly through interactions with proteins that bind the silencer DNA directly, such as the origin recognition complex (ORC). Eight recessive alleles of SIR1 were discovered that encode mutant Sir1 proteins specifically defective in their ability to recognize the HMR-E silencer. The eight missense mutations all map within a 17-amino-acid segment of Sir1p, and this segment was also required for Sir1p's interaction with Orc1p. The mutant Sir1 proteins could function in silencing if tethered to a silencer directly through a heterologous DNA-binding domain. Thus the amino acids identified are required for Sir1 protein's recognition of the HMR-E silencer and interaction with Orc1p, but not for its ability to function in silencing per se. The approach used to find these mutations may be applicable to defining interaction surfaces on proteins involved in other processes that require the assembly of macromolecular complexes. PMID- 9872945 TI - Genetic diversity of ospC in a local population of Borrelia burgdorferi sensu stricto. AB - The outer surface protein, OspC, is highly variable in Borrelia burgdorferi sensu stricto, the agent of Lyme disease. We have shown that even within a single population OspC is highly variable. The variation of ospA and ospC in the 40 infected deer ticks collected from a single site on Shelter Island, New York, was determined using PCR-SSCP. There is very strong apparent linkage disequilibrium between ospA and ospC alleles, even though they are located on separate plasmids. Thirteen discernible SSCP mobility classes for ospC were identified and the DNA sequence for each was determined. These sequences, combined with 40 GenBank sequences, allow us to define 19 major ospC groups. Sequences within a major ospC group are, on average, <1% different from each other, while sequences between major ospC groups are, on average, approximately 20% different. The tick sample contains 11 major ospC groups, GenBank contains 16 groups, with 8 groups found in both samples. Thus, the ospC variation within a local population is almost as great as the variation of a similar-sized sample of the entire species. The Ewens Watterson-Slatkin test of allele frequency showed significant deviation from the neutral expectation, indicating balancing selection for these major ospC groups. The variation represented by major ospC groups needs to be considered if the OspC protein is to be used as a serodiagnostic antigen or a vaccine. PMID- 9872947 TI - The MSN1 and NHP6A genes suppress SWI6 defects in Saccharomyces cerevisiae. AB - Ankyrin (ANK) repeats were first found in the Swi6 transcription factor of Saccharomyces cerevisiae and since then were identified in many proteins of eukaryotes and prokaryotes. These repeats are thought to serve as protein association domains. In Swi6, ANK repeats affect DNA binding of both the Swi4/Swi6 and Mbp1/Swi6 complexes. We have previously described generation of random mutations within the ANK repeats of Swi6 that render the protein temperature sensitive in its ability to activate HO transcription. Two of these SWI6 mutants were used in a screen for high copy suppressors of this phenotype. We found that MSN1, which encodes a transcriptional activator, and NHP6A, which encodes an HMG-like protein, are able to suppress defective Swi6 function. Both of these gene products are involved in HO transcription, and Nhp6A may also be involved in CLN1 transcription. Moreover, because overexpression of NHP6A can suppress caffeine sensitivity of one of the SWI6 ANK mutants, swi6-405, other SWI6-dependent genes may also be affected by Nhp6A. We hypothesize that Nhp6A and Msn1 modulate Swi6-dependent gene transcription indirectly, through effects on chromatin structure or other transcription factors, because we have not been able to demonstrate that either Msn1 or Nhp6A interact with the Swi4/Swi6 complex. PMID- 9872948 TI - Saccharomyces cerevisiae Mod5p-II contains sequences antagonistic for nuclear and cytosolic locations. AB - MOD5 encodes a tRNA modification activity located in three subcellular compartments. Alternative translation initiation generates Mod5p-I, located in the mitochondria and the cytosol, and Mod5p-II, located in the cytosol and nucleus. Here we study the nucleus/cytosol distribution of overexpressed Mod5p II. Nuclear Mod5p-II appears concentrated in the nucleolus, perhaps indicating that the nuclear pool may have a different biological role than the cytoplasmic and mitochondrial pools. Mod5p contains three motifs resembling bipartite-like nuclear localization sequences (NLSs), but only one is sufficient to locate a passenger protein to the nucleus. Mutations of basic residues of this motif cumulatively contribute to a cytosolic location for the fusion proteins. These alterations also cause decreased nuclear pools of endogenous Mod5p-II. Depletion of nuclear Mod5p-II does not affect tRNATyr function. Despite the NLS, most Mod5p is cytosolic. We assessed whether Mod5p sequences cause a karyophilic reporter to be located in the cytosol. By this assay, Mod5p may contain more than one region that functions as cytoplasmic retention and/or nuclear export sequences. Thus, distribution of Mod5p results from the presence/absence of mitochondrial targeting information and sequences antagonistic for nuclear and cytosolic locations. Mod5p is highly conserved; sequences responsible for subcellular distribution appear to reside in "accessory" motifs missing from prokaryotic counterparts. PMID- 9872949 TI - Unpredictable fitness transitions between haploid and diploid strains of the genetically loaded yeast Saccharomyces cerevisiae. AB - Mutator strains of yeast were used to accumulate random point mutations. Most of the observed changes in fitness were negative and relatively small, although major decreases and increases were also present. The average fitness of haploid strains was lowered by approximately 25% due to the accumulated genetic load. The impact of the load remained basically unchanged when a homozygous diploid was compared with the haploid from which it was derived. In other experiments a heterozygous diploid was compared with the two different loaded haploids from which it was obtained. The fitness of such a loaded diploid was much less reduced and did not correlate with the average fitness of the two haploids. There was a fitness correlation, however, when genetically related heterozygous diploids were compared, indicating that the fitness effects of the new alleles were not entirely lost in the heterozygotes. It is argued here that to explain the observed pattern of fitness transitions it is necessary to invoke nonadditive genetic interactions that go beyond the uniform masking effect of wild-type alleles. Thus, the results gathered with haploids and homozygotes should be extrapolated to heterozygotes with caution when multiple loci contribute to the genetic load. PMID- 9872950 TI - Propagation of a novel cytoplasmic, infectious and deleterious determinant is controlled by translational accuracy in Podospora anserina. AB - Some mutant strains of the filamentous fungus Podospora anserina spontaneously present a growth impairment, which has been called Crippled Growth (CG). CG is caused by a cytoplasmic and infectious factor, C. C is efficiently transmitted during mitosis but is not transmitted to the progeny after sexual crosses. C is induced by stationary phase and cured by various means, most of which stress the cells. Translational accuracy is shown to tightly regulate the propagation of C during the active growth period, because its propagation in dividing hyphae is restricted to cells that display an increased translational accuracy. However, induction of C in stationary phase proceeds independently from the translational accuracy status of the strain. CG does not seem to be accompanied by mitochondrial DNA modifications, although C activates the action of the Determinant of Senescence, another cytoplasmic and infectious element, which causes a disorganization of the mitochondrial genome. In addition, presence of C drastically modifies the spectrum of the mitochondrial DNA rearrangements in AS6 5 mat- cultures during Senescence. C seems to belong to the growing list of unconventional genetic elements. The biological significance of such elements is discussed. PMID- 9872951 TI - Extragenic suppressors of loss-of-function mutations in the aspergillus FlbA regulator of G-protein signaling domain protein. AB - We showed previously that two genes, fl bA and fadA, have a major role in determining the balance between growth, sporulation, and mycotoxin (sterigmatocystin; ST) production by the filamentous fungus Aspergillus nidulans. fadA encodes the alpha subunit for a heterotrimeric G-protein, and continuous activation of FadA blocks sporulation and ST production while stimulating growth. fl bA encodes an A. nidulans regulator of G-protein signaling (RGS) domain protein that antagonizes FadA-mediated signaling to allow development. To better understand FlbA function and other aspects of FadA-mediated growth control, we have isolated and characterized mutations in four previously undefined genes designated as sfaA, sfaC, sfaD, and sfaE (suppressors of flbA), and a new allele of fadA (fadAR205H), all of which suppress a fl bA loss-of-function mutation ( fl bA98). These suppressors overcome fl bA losses of function in both sporulation and ST biosynthesis. fadAR205H, sfaC67, sfaD82, and sfaE83 mutations are dominant to wild type whereas sfaA1 is semidominant. sfaA1 also differs from other suppressor mutations in that it cannot suppress a fl bA deletion mutation (and is therefore allele specific) whereas all the dominant suppressors can bypass complete loss of fl bA. Only sfaE83 suppressed dominant activating mutations in fadA, indicating that sfaE may have a unique role in fadA- fl bA interactions. Finally, none of these suppressor mutations bypassed fl uG loss-of-function mutations in development-specific activation. PMID- 9872952 TI - Mutational activation of a Galphai causes uncontrolled proliferation of aerial hyphae and increased sensitivity to heat and oxidative stress in Neurospora crassa. AB - Heterotrimeric G proteins, consisting of alpha, beta, and gamma subunits, transduce environmental signals through coupling to plasma membrane-localized receptors. We previously reported that the filamentous fungus Neurospora crassa possesses a Galpha protein, GNA-1, that is a member of the Galphai superfamily. Deletion of gna-1 leads to defects in apical extension, differentiation of asexual spores, sensitivity to hyperosmotic media, and female fertility. In addition, Deltagna-1 strains have lower intracellular cAMP levels under conditions that promote morphological abnormalities. To further define the function of GNA-1 in signal transduction in N. crassa, we examined properties of strains with mutationally activated gna-1 alleles (R178C or Q204L) as the only source of GNA-1 protein. These mutations are predicted to inhibit the GTPase activity of GNA-1 and lead to constitutive signaling. In the sexual cycle, gna 1(R178C) and gna-1(Q204L) strains are female-fertile, but produce fewer and larger perithecia than wild type. During asexual development, gna-1(R178C) and gna-1(Q204L) strains elaborate abundant, long aerial hyphae, produce less conidia, and possess lower levels of carotenoid pigments in comparison to wild type controls. Furthermore, gna-1(R178C) and gna-1(Q204L) strains are more sensitive to heat shock and exposure to hydrogen peroxide than wild-type strains, while Deltagna-1 mutants are more resistant. In contrast to Deltagna-1 mutants, gna-1(R178C) and gna-1(Q204L) strains have higher steady-state levels of cAMP than wild type. The results suggest that GNA-1 possesses several Gbetagamma independent functions in N. crassa. We propose that GNA-1 mediates signal transduction pathway(s) that regulate aerial hyphae development and sensitivity to heat and oxidative stresses, possibly through modulation of cAMP levels. PMID- 9872953 TI - The rate of spontaneous mutation for life-history traits in Caenorhabditis elegans. AB - Spontaneous mutations were accumulated in 100 replicate lines of Caenorhabditis elegans over a period of approximately 50 generations. Periodic assays of these lines and comparison to a frozen control suggest that the deleterious mutation rate for typical life-history characters in this species is at least 0.05 per diploid genome per generation, with the average mutational effect on the order of 14% or less in the homozygous state and the average mutational heritability approximately 0.0034. While the average mutation rate per character and the average mutational heritability for this species are somewhat lower than previous estimates for Drosophila, these differences can be reconciled to a large extent when the biological differences between these species are taken into consideration. PMID- 9872954 TI - Complexity of developmental control: analysis of embryonic cell lineage specification in Caenorhabditis elegans using pes-1 as an early marker. AB - In the early Caenorhabditis elegans embryo five somatic founder cells are born during the first cleavages. The first of these founder cells, named AB, gives rise to 389 of the 558 nuclei present in the hatching larva. Very few genes directly involved in the specification of the AB lineage have been identified so far. Here we describe a screen of a large collection of maternal-effect embryonic lethal mutations for their effect on the early expression of a pes-1::lacZ fusion gene. This fusion gene is expressed in a characteristic pattern in 14 of the 32 AB descendants present shortly after the initiation of gastrulation. Of the 37 mutations in 36 genes suspected to be required specifically during development, 12 alter the expression of the pes-1::lacZ marker construct. The gene expression pattern alterations are of four types: reduction of expression, variable expression, ectopic expression in addition to the normal pattern, and reduction of the normal pattern together with ectopic expression. We estimate that approximately 100 maternal functions are required to establish the pes-1 expression pattern in the early embryo. PMID- 9872956 TI - The Amylase gene cluster on the evolving sex chromosomes of Drosophila miranda. AB - On the basis of chromosomal homology, the Amylase gene cluster in Drosophila miranda must be located on the secondary sex chromosome pair, neo-X (X2) and neo Y, but is autosomally inherited in all other Drosophila species. Genetic evidence indicates no active amylase on the neo-Y chromosome and the X2-chromosomal locus already shows dosage compensation. Several lines of evidence strongly suggest that the Amy gene cluster has been lost already from the evolving neo-Y chromosome. This finding shows that a relatively new neo-Y chromosome can start to lose genes and hence gradually lose homology with the neo-X. The X2 chromosomal Amy1 is intact and Amy2 contains a complete coding sequence, but has a deletion in the 3'-flanking region. Amy3 is structurally eroded and hampered by missing regulatory motifs. Functional analysis of the X2-chromosomal Amy1 and Amy2 regions from D. miranda in transgenic D. melanogaster flies reveals ectopic AMY1 expression. AMY1 shows the same electrophoretic mobility as the single amylase band in D. miranda, while ectopic AMY2 expression is characterized by a different mobility. Therefore, only the Amy1 gene of the resident Amy cluster remains functional and hence Amy1 is the dosage compensated gene. PMID- 9872955 TI - The levels of the RoRNP-associated Y RNA are dependent upon the presence of ROP 1, the Caenorhabditis elegans Ro60 protein. AB - The Ro ribonucleoproteins (RoRNP) consist of at least one major protein of 60 kD, Ro60, and one small associated RNA, designated Y RNA. Although RoRNP have been found in all vertebrate species examined so far, their function remains unknown. The Caenorhabditis elegans rop-1 gene previously has been identified as encoding a Ro60 homologue. We report here the phenotypic characterization of a C. elegans strain in which rop-1 has been disrupted. This is the first report regarding the inactivation of a major RoRNP constituent in any organism. The rop-1 mutant worms display no visible defects. However, at the molecular level, the disruption of rop-1 results in a dramatic decrease in the levels of the ROP-1-associated RNA (CeY RNA). Moreover, transgenic expression of wild-type rop-1 partially rescues the levels of CeY RNA. Considering that transgenes are poorly expressed in the germline, the fact that the rescue is only partial is most likely related to the high abundance of the CeY RNA in the adult germline and in embryos. The developmental expression pattern and localization of CeY RNA suggest a role for this molecule during embryogenesis. We conclude that, under laboratory culture conditions, ROP-1 does not play a crucial role in C. elegans. PMID- 9872957 TI - The haplolethal region at the 16F gene cluster of Drosophila melanogaster: structure and function. AB - Extensive aneuploid analyses had shown the existence of a few haplolethal (HL) regions and one triplolethal region in the genome of Drosophila melanogaster. Since then, only two haplolethals, 22F1-2 and 16F, have been directly linked to identified genes, dpp and wupA, respectively. However, with the possible exception of dpp, the actual bases for this dosage sensitivity remain unknown. We have generated and characterized dominant-lethal mutations and chromosomal rearrangements in 16F and studied them in relation to the genes in the region. This region extends along 100 kb and includes at least 14 genes. The normal HL function depends on the integrity of a critical 4-kb window of mostly noncoding sequences within the wupA transcription unit that encodes the muscle protein troponin I (TNI). All dominant lethals are breakpoints within that window, which prevent the functional expression of TNI and other adjacent genes in the proximal direction. However, independent mutations in these genes result in recessive lethal phenotypes only. We propose that the HL at 16F represents a long-range cis regulatory region that acts upon a number of functionally related genes whose combined haploidy would yield the dominant-lethal effect. PMID- 9872958 TI - Local transposition of a hobo element within the decapentaplegic locus of Drosophila. AB - We have efficiently mobilized a phenotypically silent hobo transgene inserted within the cis-regulatory heldout region of the decapentaplegic (dpp) locus in Drosophila melanogaster. The goal of our experiment was to identify germline transmission of a local transposition event within the dpp locus that meets two specific criteria. First, excision of the hobo construct does not generate an adult mutant phenotype, suggesting minimal alteration to the original site of insertion. Second, we required a new insertion of the hobo transgene into the Haploinsufficient region of the locus approximately 25 kb away. Genetic and molecular criteria are used to evaluate candidate germlines. In a pilot study, this local transposition event occurred independently in two individuals. Both of the transposition events appear to be new insertions into the dpp transcription unit. One insertion is between the two protein-coding exons, and the other is in the 3'-untranslated region of exon three. Strains carrying these insertions are valuable new reagents for the analysis of dpp function and molecular evolution. These results further support the use of the hobo system as an important tool in Drosophila genetics. PMID- 9872960 TI - Organization of the large mitochondrial genome in the isopod Armadillidium vulgare. AB - The mitochondrial DNA (mtDNA) in animals is generally a circular molecule of approximately 15 kb, but there are many exceptions such as linear molecules and larger ones. RFLP studies indicated that the mtDNA in the terrestrial isopod Armadillidium vulgare varied from 20 to 42 kb. This variation depended on the restriction enzyme used, and on the restriction profile generated by a given enzyme. The DNA fragments had characteristic electrophoretic behaviors. Digestions with two endonucleases always generated fewer fragments than expected; denaturation of restriction profiles reduced the size of two bands by half; densitometry indicated that a number of small fragments were present in stoichiometry, which has approximately twice the expected concentration. Finally, hybridization to a 550-bp 16S rDNA probe often revealed two copies of this gene. These results cannot be due to the genetic rearrangements generally invoked to explain large mtDNA. We propose that the large A. vulgare mtDNA is produced by the tripling of a 14-kb monomer with a singular rearrangement: one monomer is linear and the other two form a circular dimer. Densitometry suggested that these two molecular structures were present in different proportions within a single individual. The absence of mutations within the dimers also suggests that replication occurs during the monomer phase. PMID- 9872959 TI - Molecular population genetics of the rp49 gene region in different chromosomal inversions of Drosophila subobscura. AB - Nucleotide variation at the ribosomal protein 49 (rp49) gene region has been studied in 75 lines of Drosophila subobscura belonging to four chromosomal arrangements (Ost, O3+4, O3+4+8, and O3+4+23). The location of the rp49 gene region within the inversion loop differs among heterokaryotypes: it is very close to one of the breakpoints in heterozygotes involving Ost chromosomes, while it is in a more central position in all other heterokaryotypes. The distribution of nucleotide polymorphism in the different arrangements is consistent with a monophyletic origin of the inversions. The data also provide evidence that gene conversion and possibly double crossover are involved in shuffling nucleotide variation among gene arrangements. The analyses reveal that the level of genetic exchange is higher when the region is located in a more central position of the inverted fragment than when it is close to the breakpoints. The pairwise difference distributions as well as the negative values of Tajima's and Fu and Li's statistics further support the hypothesis that nucleotide variation within chromosomal arrangements still reflects expansion after the origin of the inversions. Under the expansion model, we have estimated the time of origin of the studied inversions. PMID- 9872961 TI - The dominant temperature-sensitive lethal DTS7 of Drosophila melanogaster encodes an altered 20S proteasome beta-type subunit. AB - Proteasomes are multicatalytic complexes that function as the major proteolytic machinery in regulated protein degradation. The eukaryotic 20S proteasome proteolytic core structure comprises 14 different subunits: 7 alpha-type and 7 beta-type. DTS7 is a dominant temperature-sensitive (DTS) lethal mutation at 29 degrees that also acts as a recessive lethal at ambient temperatures. DTS7 maps to cytological position 71AB. Molecular characterization of DTS7 reveals that this is caused by a missense mutation in a beta-type subunit gene, beta2. A previously characterized DTS mutant, l(3)73Ai1, results from a missense mutation in another beta-type subunit gene, beta6. These two mutants share a very similar phenotype, show a strong allele-specific genetic interaction, and are rescued by the same extragenic suppressor, Su(DTS)-1. We propose that these mutants might act as "poison subunits," disrupting proteasome function in a dosage-dependent manner, and suggest how they may interact on the basis of the structure of the yeast 20S proteasome. PMID- 9872962 TI - Inferring the fitness effects of DNA mutations from polymorphism and divergence data: statistical power to detect directional selection under stationarity and free recombination. AB - The fitness effects of classes of DNA mutations can be inferred from patterns of nucleotide variation. A number of studies have attributed differences in levels of polymorphism and divergence between silent and replacement mutations to the action of natural selection. Here, I investigate the statistical power to detect directional selection through contrasts of DNA variation among functional categories of mutations. A variety of statistical approaches are applied to DNA data simulated under Sawyer and Hartl's Poisson random field model. Under assumptions of free recombination and stationarity, comparisons that include both the frequency distributions of mutations segregating within populations and the numbers of mutations fixed between populations have substantial power to detect even very weak selection. Frequency distribution and divergence tests are applied to silent and replacement mutations among five alleles of each of eight Drosophila simulans genes. Putatively "preferred" silent mutations segregate at higher frequencies and are more often fixed between species than "unpreferred" silent changes, suggesting fitness differences among synonymous codons. Amino acid changes tend to be either rare polymorphisms or fixed differences, consistent with a combination of deleterious and adaptive protein evolution. In these data, a substantial fraction of both silent and replacement DNA mutations appear to affect fitness. PMID- 9872964 TI - Are unpaired chromosomes spermicidal?: A maximum-likelihood analysis of segregation and meiotic drive in Drosophila melanogaster males deficient for the ribosomal-dna. AB - Meiosis in Drosophila melanogaster males is achiasmate and requires special systems to ensure normal segregation. Several situations that yield frequent nondisjunction also produce high levels of chromatin-dependent sperm lethality, suggesting the possibility of a simple and direct connection between defective disjunction and defective sperm development. One hypothesis that has been offered is that pairing not only ensures disjunction, but also changes the physical state of chromosomes so that they can be packaged in sperm. Here, I present an analysis of extensive data on disjunction and sperm survival in rDNA-deficient males collected by B. McKee and D. Lindsley. This analysis demonstrates that, although nondisjunction and sperm lethality are indeed correlated, the basis of this is not the presence of unpaired chromosomes in the sperm. Chromosomes that have failed to disjoin are not themselves spermicidal. PMID- 9872963 TI - Natural selection on synonymous sites is correlated with gene length and recombination in Drosophila. AB - Evolutionary analysis of codon bias in Drosophila indicates that synonymous mutations are not neutral, but rather are subject to weak selection at the translation level. Here we show that the effectiveness of natural selection on synonymous sites is strongly correlated with the rate of recombination, in accord with the nearly neutral hypothesis. This correlation, however, is apparent only in genes encoding short proteins. Long coding regions have both a lower codon bias and higher synonymous substitution rates, suggesting that they are affected less efficiently by selection. Therefore, both the length of the coding region and the recombination rate modulate codon bias. In addition, the data indicate that selection coefficients for synonymous mutations must vary by a minimum of one or two orders of magnitude. Two hypotheses are proposed to explain the relationship among the coding region length, the codon bias, and the synonymous divergence and polymorphism levels across the range of recombination rates in Drosophila. The first hypothesis is that selection coefficients on synonymous mutations are inversely related to the total length of the coding region. The second hypothesis proposes that interference among synonymous mutations reduces the efficacy of selection on these mutations. We investigated this second hypothesis by carrying out forward simulations of weakly selected mutations in model populations. These simulations show that even with realistic recombination rates, this interference, which we call the "small-scale" Hill-Robertson effect, can have a moderately strong influence on codon bias. PMID- 9872966 TI - Production of fertile unreduced sperm by hybrid males of the Rutilus alburnoides complex (Teleostei, cyprinidae). An alternative route to genome tetraploidization in unisexuals. AB - The hybrid minnow Rutilus alburnoides comprises diploid and polyploid females and males. Previous studies revealed that diploid and triploid females exhibit altered oogenesis that does not involve random segregation and recombination of the genomes of the two ancestors, constituting unisexual lineages. In the present study, we investigated the reproductive mode of hybrid males from the Tejo basin, using experimental crosses and flow cytometric analysis of blood and sperm. The results suggest that diploid hybrids produced fertile unreduced sperm, transmitting their hybrid genome intact to offspring. Triploid hybrids also produced unreduced sperm, but it was not possible to obtain data concerning their fertility. Finally, tetraploid hybrids produced fertile diploid sperm, which exhibited Mendelian segregation. Tetraploid R. alburnoides may reestablish biparental reproduction, as individuals of both sexes with the appropriate constitution for normal meiosis (two haploid genomes from each parental species) are likely to occur in natural populations. Tetraploids probably have arisen from syngamy of diploid eggs and diploid sperm produced by diploid hybrid males. Diploid hybrid males may therefore play a significant role in the dynamics of the complex, starting the evolutionary process that may ultimately lead to a new sexually reproducing species. PMID- 9872965 TI - The role of evolutionarily conserved sequences in alternative splicing at the 3' end of Drosophila melanogaster myosin heavy chain RNA. AB - Exon 18 of the muscle myosin heavy chain gene (Mhc) of Drosophila melanogaster is excluded from larval transcripts but included in most adult transcripts. To identify cis-acting elements regulating this alternative RNA splicing, we sequenced the 3' end of Mhc from the distantly related species D. virilis. Three noncoding regions are conserved: (1) the nonconsensus splice junctions at either end of exon 18; (2) exon 18 itself; and (3) a 30-nucleotide, pyrimidine-rich sequence located about 40 nt upstream of the 3' splice site of exon 18. We generated transgenic flies expressing Mhc mini-genes designed to test the function of these regions. Improvement of both splice sites of adult-specific exon 18 toward the consensus sequence switches the splicing pattern to include exon 18 in all larval transcripts. Thus nonconsensus splice junctions are critical to stage-specific exclusion of this exon. Deletion of nearly all of exon 18 does not affect stage-specific utilization. However, splicing of transcripts lacking the conserved pyrimidine sequence is severely disrupted in adults. Disruption is not rescued by insertion of a different polypyrimidine tract, suggesting that the conserved pyrimidine-rich sequence interacts with tissue specific splicing factors to activate utilization of the poor splice sites of exon 18 in adult muscle. PMID- 9872968 TI - Time-related mapping of quantitative trait loci underlying tiller number in rice. AB - Using time-related phenotypic data, methods of composite interval mapping and multiple-trait composite interval mapping based on least squares were applied to map quantitative trait loci (QTL) underlying the development of tiller number in rice. A recombinant inbred population and a corresponding saturated molecular marker linkage map were constructed for the study. Tiller number was recorded every 4 or 5 days for a total of seven times starting at 20 days after sowing. Five QTL were detected on chromosomes 1, 3, and 5. These QTL explained more than half of the genetic variance at the final observation. All the QTL displayed an S shaped expression curve. Three QTL reached their highest expression rates during active tillering stage, while the other two QTL achieved this either before or after the active tillering stage. PMID- 9872967 TI - Multiple levels of single-strand slippage at cetacean tri- and tetranucleotide repeat microsatellite loci. AB - Between three and six tri- and tetranucleotide repeat microsatellite loci were analyzed in 3720 samples collected from four different species of baleen whales. Ten of the 18 species/locus combinations had imperfect allele arrays, i.e., some alleles differed in length by other than simple integer multiples of the basic repeat length. The estimate of the average number of alleles and heterozygosity was higher at loci with imperfect allele arrays relative to those with perfect allele arrays. Nucleotide sequences of 23 different alleles at one tetranucleotide repeat microsatellite locus in fin whales, Balaenoptera physalus, and humpback whales, Megaptera novaeangliae, revealed sequence changes including perfect repeats only, multiple repeats, and partial repeats. The relative rate of the latter two categories of mutation was estimated at 0.024 of the mutation rate involving perfect repeats only. It is hypothesized that single-strand slippage of partial repeats may provide a mechanism for counteracting the continuous expansion of microsatellite loci, which is the logical consequence of recent reports demonstrating directional mutations. Partial-repeat mutations introduce imperfections in the repeat array, which subsequently could reduce the rate of single-strand slippage. Limited computer simulations confirmed this predicted effect of partial-repeat mutations. PMID- 9872970 TI - A consensus map for loblolly pine (Pinus taeda L.). I. Construction and integration of individual linkage maps from two outbred three-generation pedigrees. AB - A consensus map for loblolly pine (Pinus taeda L.) was constructed from the integration of linkage data from two unrelated three-generation outbred pedigrees. The progeny segregation data from restriction fragment length polymorphism, random amplified polymorphic DNA, and isozyme genetic markers from each pedigree were recoded to reflect the two independent populations of parental meioses, and genetic maps were constructed to represent each parent. The rate of meiotic recombination was significantly greater for males than females, as was the average estimate of genome length for males (1983.7 cM [Kosambi mapping function (K)]) and females [1339.5 cM(K)]. The integration of individual maps allows for the synthesis of genetic information from independent sources onto a single consensus map and facilitates the consolidation of linkage groups to represent the chromosomes n = 12 of loblolly pine. The resulting consensus map consists of 357 unique molecular markers and covers approximately 1300 cM(K). PMID- 9872969 TI - Suppressors of the arabidopsis lsd5 cell death mutation identify genes involved in regulating disease resistance responses. AB - Cell death is associated with the development of the plant disease resistance hypersensitive reaction (HR). Arabidopsis lsd mutants that spontaneously exhibit cell death reminiscent of the HR were identified previously. To study further the regulatory context in which cell death acts during disease resistance, one of these mutants, lsd5, was used to isolate new mutations that suppress its cell death phenotype. Using a simple lethal screen, nine lsd5 cell death suppressors, designated phx (for the mythological bird Phoenix that rises from its ashes), were isolated. These mutants were characterized with respect to their response to a bacterial pathogen and oomycete parasite. The strongest suppressors-phx2, 3, 6, and 11-1-showed complex, differential patterns of disease resistance modifications. These suppressors attenuated disease resistance to avirulent isolates of the biotrophic Peronospora parasitica pathogen, but only phx2 and phx3 altered disease resistance to avirulent strains of Pseudomonas syringae pv tomato. Therefore, some of these phx mutants define common regulators of cell death and disease resistance. In addition, phx2 and phx3 exhibited enhanced disease susceptibility to different virulent pathogens, confirming probable links between the disease resistance and susceptibility pathways. PMID- 9872971 TI - Functional analysis of deletion derivatives of the maize transposon MuDR delineates roles for the MURA and MURB proteins. AB - The regulatory transposon of the Mutator system of transposable elements in maize is MuDR. MuDR elements produce two transcripts, from genes mudrA and mudrB, encoding proteins MURA and MURB, respectively. Like many other transposons, MuDR elements often undergo deletions, usually of internal sequences. Analysis of a deletion that is restricted to the region encoding MURB demonstrates that this gene is not required to cause excisions of a reporter element, although it may be required for transposition or suppression of suppressible alleles. Conversely, a derivative that lacks the region encoding MURA but that produces MURB is nonfunctional for all aspects of Mutator activity. Northern analysis of these derivatives reveals that each of the two transcripts can be independently transcribed, and analysis using an antibody specific for MURB reveals that mudrB transcript can also be successfully translated and its product appropriately localized in the absence of mudrA. A third deletion derivative provides evidence for a source of previously reported antisense transcript. PMID- 9872973 TI - Estimating linkage disequilibrium between a polymorphic marker locus and a trait locus in natural populations. AB - Positional cloning of gene(s) underlying a complex trait requires a high resolution linkage map between the trait locus and genetic marker loci. Recent research has shown that this may be achieved through appropriately modeling and screening linkage disequilibrium between the candidate marker locus and the major trait locus. A quantitative genetics model was developed in the present study to estimate the coefficient of linkage disequilibrium between a polymorphic genetic marker locus and a locus underlying a quantitative trait as well as the relevant genetic parameters using the sample from randomly mating populations. Asymptotic covariances of the maximum-likelihood estimates of the parameters were formulated. Convergence of the EM-based statistical algorithm for calculating the maximum-likelihood estimates was confirmed and its utility to analyze practical data was exploited by use of extensive Monte-Carlo simulations. Appropriateness of calculating the asymptotic covariance matrix in the present model was investigated for three different approaches. Numerical analyses based on simulation data indicated that accurate estimation of the genetic parameters may be achieved if a sample size of 500 is used and if segregation at the trait locus explains not less than a quarter of phenotypic variation of the trait, but the study reveals difficulties in predicting the asymptotic variances of these maximum-likelihood estimates. A comparison was made between the statistical powers of the maximum-likelihood analysis and the previously proposed regression analysis for detecting the disequilibrium. PMID- 9872972 TI - The effect of mating system differences on nucleotide diversity at the phosphoglucose isomerase locus in the plant genus Leavenworthia. AB - To test the theoretical prediction that highly inbreeding populations should have low neutral genetic diversity relative to closely related outcrossing populations, we sequenced portions of the cytosolic phosphoglucose isomerase (PgiC) gene in the plant genus Leavenworthia, which includes both self incompatible and inbreeding taxa. On the basis of sequences of intron 12 of this gene, the expected low diversity was seen in both populations of the selfers Leavenworthia uniflora and L. torulosa and in three highly inbreeding populations of L. crassa, while high diversity was found in self-incompatible L. stylosa, and moderate diversity in L. crassa populations with partial or complete self incompatibility. In L. stylosa, the nucleotide diversity was strongly structured into three haplotypic classes, differing by several insertion/deletion sequences, with linkage disequilibrium between sequences of the three types in intron 12, but not in the adjacent regions. Differences between the three kinds of haplotypes are larger than between sequences of this gene region from different species. The haplotype divergence suggests the presence of a balanced polymorphism at this locus, possibly predating the split between L. stylosa and its two inbreeding sister taxa, L. uniflora and L. torulosa. It is therefore difficult to distinguish between different potential causes of the much lower sequence diversity at this locus in inbreeding than outcrossing populations. Selective sweeps during the evolution of these populations are possible, or background selection, or merely loss of a balanced polymorphism maintained by overdominance in the populations that evolved high selfing rates. PMID- 9872974 TI - Statistical methods for mapping quantitative trait loci from a dense set of markers. AB - Lander and Botstein introduced statistical methods for searching an entire genome for quantitative trait loci (QTL) in experimental organisms, with emphasis on a backcross design and QTL having only additive effects. We extend their results to intercross and other designs, and we compare the power of the resulting test as a function of the magnitude of the additive and dominance effects, the sample size and intermarker distances. We also compare three methods for constructing confidence regions for a QTL: likelihood regions, Bayesian credible sets, and support regions. We show that with an appropriate evaluation of the coverage probability a support region is approximately a confidence region, and we provide a theroretical explanation of the empirical observation that the size of the support region is proportional to the sample size, not the square root of the sample size, as one might expect from standard statistical theory. PMID- 9872975 TI - Single- and multiple-trait mapping analysis of linked quantitative trait loci. Some asymptotic analytical approximations. AB - Estimating the resolution power of mapping analysis of linked quantitative trait loci (QTL) remains a difficult problem, which has been previously addressed mainly by Monte Carlo simulations. The analytical method of evaluation of the expected LOD developed in this article spreads the "deterministic sampling" approach for the case of two linked QTL for single- and two-trait analysis. Several complicated questions are addressed through this evaluation: the dependence of QTL detection power on the QTL effects, residual correlation between the traits, and the effect of epistatic interaction between the QTL for one or both traits on expected LOD (ELOD), etc. Although this method gives only an asymptotic estimation of ELOD, it allows one to get an approximate assessment of a broad spectrum of mapping situations. A good correspondence was found between the ELODs predicted by the model and LOD values averaged over Monte Carlo simulations. PMID- 9872976 TI - Genetic differentiation within and between two habitats. AB - We investigate the usefulness of analyses of population differentiation between different ecological types, such as host races of parasites or sources and sink habitats. To that aim, we formulate a model of population structure involving two classes of subpopulations found in sympatry. Extensions of previous results for Wright's F-statistics in island and isolation-by-distance models of dispersal are given. It is then shown that source and sinks cannot in general be distinguished by F-statistics nor by their gene diversities. The excess differentiation between two partially isolated classes with respect to differentiation within classes is shown to decrease with distance, and for a wide range of parameter values it should be difficult to detect. In the same circumstances little differentiation will be observed in "hierarchical" analyses between pools of samples from each habitat, and differences between levels of differentiation within each habitat will only reflect differences between levels of gene diversity within each habitat. Exceptions will indicate strong isolation between the different classes or habitat-related divergent selection. PMID- 9872977 TI - Detection of quantitative trait loci in outbred populations with incomplete marker data. AB - Augmentation of marker genotypes for ungenotyped individuals is implemented in a Bayesian approach via the use of Markov chain Monte Carlo techniques. Marker data on relatives and phenotypes are combined to compute conditional posterior probabilities for marker genotypes of ungenotyped individuals. The presented procedure allows the analysis of complex pedigrees with ungenotyped individuals to detect segregating quantitative trait loci (QTL). Allelic effects at the QTL were assumed to follow a normal distribution with a covariance matrix based on known QTL position and identity by descent probabilities derived from flanking markers. The Bayesian approach estimates variance due to the single QTL, together with polygenic and residual variance. The method was empirically tested through analyzing simulated data from a complex granddaughter design. Ungenotyped dams were related to one or more sons or grandsires in the design. Heterozygosity of the marker loci and size of QTL were varied. Simulation results indicated a significant increase in power when ungenotyped dams were included in the analysis. PMID- 9872978 TI - A DNA polymorphism discovery resource for research on human genetic variation. PMID- 9872979 TI - Models of molecular evolution and phylogeny. AB - Phylogenetic reconstruction is a fast-growing field that is enriched by different statistical approaches and by findings and applications in a broad range of biological areas. Fundamental to these are the mathematical models used to describe the patterns of DNA base substitution and amino acid replacement. These may become some of the basic models for comparative genome research. We discuss these models, including the analysis of observed DNA base and amino acid mutation patterns, the concept of site heterogeneity, and the incorporation of structural biology data, all of which have become particularly important in recent years. We also describe the use of such models in phylogenetic reconstruction and statistical methods for the comparison of different models. PMID- 9872980 TI - Strategies for mutational analysis of the large multiexon ATM gene using high density oligonucleotide arrays. AB - Mutational analysis of large genes with complex genomic structures plays an important role in medical genetics. Technical limitations associated with current mutation screening protocols have placed increased emphasis on the development of new technologies to simplify these procedures. High-density arrays of >90,000 oligonucleotide probes, 25 nucleotides in length, were designed to screen for all possible heterozygous germ-line mutations in the 9.17-kb coding region of the ATM gene. A strategy for rapidly developing multiexon PCR amplification protocols in DNA chip-based hybridization analysis was devised and implemented in preparing target for the 62 ATM coding exons. Improved algorithms for interpreting data from two-color experiments, where reference and test samples are cohybridized to the arrays, were developed. In a blinded study, 17 of 18 distinct heterozygous and 8 of 8 distinct homozygous sequence variants in the assayed region were detected accurately along with five false-positive calls while scanning >200 kb in 22 genomic DNA samples. Of eight heterozygous sequence changes found in more than one sample, six were detected in all cases. Five previously unreported sequence changes, not found by other mutational scanning methodologies on these same samples, were detected that led to either amino acid changes or premature truncation of the ATM protein. DNA chip-based assays should play a valuable role in high throughput sequence analysis of complex genes. PMID- 9872981 TI - Homologs of the yeast longevity gene LAG1 in Caenorhabditis elegans and human. AB - LAG1 is a longevity gene, the first such gene to be identified and cloned from the yeast Saccharomyces cerevisiae. A close homolog of this gene, which we call LAC1, has been found in the yeast genome. We have cloned the human homolog of LAG1 with the ultimate goal of examining its possible function in human aging. In the process, we have also cloned a homolog from the nematode worm Caenorhabditis elegans. Both of these homologs, LAG1Hs and LAG1Ce-1, functionally complemented the lethality of a lag1delta lac1delta double deletion, despite low overall sequence similarity to the yeast proteins. The proteins shared a short sequence, the Lag1 motif, and a similar transmembrane domain profile. Another, more distant human homolog, TRAM, which lacks this motif, did not complement. LAG1Hs also restored the life span of the double deletion, demonstrating that it functions in establishing the longevity phenotype in yeast. LAG1Hs mapped to 19p12, and it was expressed in only three tissues: brain, skeletal muscle, and testis. This gene possesses a trinucleotide (CTG) repeat within exon 1. This and its expression profile raise the possibility that it may be involved in neurodegenerative disease. This possibility suggests at least one way in which LAG1Hs might be involved in human aging. PMID- 9872982 TI - The relative power of family-based and case-control designs for linkage disequilibrium studies of complex human diseases I. DNA pooling. AB - We consider statistics for analyzing a variety of family-based and nonfamily based designs for detecting linkage disequilibrium of a marker with a disease susceptibility locus. These designs include sibships with parents, sibships without parents, and use of unrelated controls. We also provide formulas for and evaluate the relative power of different study designs using these statistics. In this first paper in the series, we derive statistical tests based on data derived from DNA pooling experiments and describe their characteristics. Although designs based on affected and unaffected sibs without parents are usually robust to population stratification, they suffer a loss of power compared with designs using parents or unrelateds as controls. Although increasing the number of unaffected sibs improves power, the increase is generally not substantial. Designs including sibships with multiple affected sibs are typically the most powerful, with any of these control groups, when the disease allele frequency is low. When the allele frequency is high, however, designs with unaffected sibs as controls do not retain this advantage. In designs with parents, having an affected parent has little impact on the power, except for rare dominant alleles, where the power is increased compared with families with no affected parents. Finally, we also demonstrate that for sibships with parents, only the parents require individual genotyping to derive the TDT statistic, whereas all the offspring can be pooled. This can potentially lead to considerable savings in genotyping, especially for multiplex sibships. The formulas and tables we derive should provide some guidance to investigators designing nuclear family-based linkage disequilibrium studies for complex diseases. PMID- 9872983 TI - MS205 minisatellite diversity in Basques: evidence for a pre-Neolithic component. AB - A number of studies have suggested that Basques might be a relic of Mesolithic Europeans who escaped much of the homogenization brought about by the Neolithic expansion. In an attempt to add new insights into this hypothesis, MS205 minisatellite diversity has been investigated by Minisatellite Variant Repeat (MVR) analysis in a sample of >100 autochthonous individuals from the Basque Country, along with 24 Castilian (N. Spain) and 23 individuals from the United Kingdom. These populations were examined in the context of the available world database for MS205 alleles. To deduce the similarities among populations, we have applied a phylogenetic approach that takes into account similarity between alleles. The variability of these populations seems to be a subset of the greater and presumably older African diversity, as has been suggested previously for non Africans. Within non-Africans, Basques seem to cluster with other Northern European populations; however, some apparently Basque-specific alleles can be dated back to post-Aurignacian times, supporting the continuity of some lineages of this population since the Upper Paleolithic period. PMID- 9872984 TI - A HAPPY map of Cryptosporidium parvum. AB - We have constructed a HAPPY map of the apicomplexan parasite Cryptosporidium parvum. We have placed 204 markers on the 10.4-Mb genome, giving an average marker spacing of approximately 50 kb, with an effective resolution of approximately 40 kb. HAPPY mapping (an in vitro linkage technique based on screening approximately haploid amounts of DNA by the polymerase chain reaction) is fast and accurate and is not subject to the distortions inherent in cloning, meiotic recombination, or hybrid cell formation. In addition, little genomic DNA is needed as a substrate, and the AT content of the genome is largely immaterial, making it an ideal method for mapping otherwise intractable parasite genomes. The map, covering all eight chromosomes, consists of 10 linkage groups, each of which has been chromosomally assigned. We have verified the accuracy of the map by several methods, including the construction of a >140-kb PAC contig on chromosome VI. Less than 1% of our markers detect non-rDNA duplicated sequences. PMID- 9872985 TI - Scriptable access to the Caenorhabditis elegans genome sequence and other ACEDB databases. AB - Much of the world's genomic data are available to the community through networked databases that are accessed via Web interfaces. Although this paradigm provides browse-level access and has greatly facilitated linking between databases, it does not provide any convenient mechanism for programmatically fetching and integrating data from diverse databases. We have created a library and an application programming interface (API) named AcePerl that provides simple, direct access to ACEDB databases from the Perl programming language. With this library, programmers and computer-savvy biologists can write software to pose complex queries on local and remote ACEDB databases, retrieve the data, integrate the results, and move data objects from one database to another. In addition, a set of Web scripts running on top of AcePerl provides Web-based browsing of any local or remote ACEDB database. AcePerl and the AceBrowser Web browser run on Unix systems and are available under a license that allows for unrestricted use and redistribution. Both packages can be downloaded from URL. A Microsoft Windows port of AcePerl is in the planning stages. PMID- 9872986 TI - Prions of yeast and fungi. Proteins as genetic material. PMID- 9872987 TI - Integrin-associated protein is a ligand for the P84 neural adhesion molecule. AB - P84 (also known as SHPS-1, BIT, and SIRP) is a heterophilic adhesive membrane protein involved in receptor tyrosine kinase signaling that is found at synapses in the mammalian central nervous system and in non-neural tissues. We have identified a binding partner for P84 using an expression cloning strategy. Here we report that integrin-associated protein (IAP/CD47) is a predominant binding partner of P84. Immunohistochemistry reveals a virtually identical distribution of P84 and IAP in a variety of adult brain regions. Because IAP has been implicated in cell signaling in cells of the immune system, P84 and IAP represent a heterophilic binding pair that is likely to be involved in bi-directional signaling at the synapse and in other tissues. PMID- 9872988 TI - Thyroglobulin type-1 domains in equistatin inhibit both papain-like cysteine proteinases and cathepsin D. AB - Equistatin from sea anemone is a protein composed of three thyroglobulin-type 1 domains known to inhibit papain-like cysteine proteinases, papain, and cathepsins B and L. Limited proteolysis was used to dissect equistatin into a first domain, eq d-1, and a combined second and third domain, eq d-2,3. Only the N-terminal domain inhibits papain (Ki = 0.61 nM). Remarkably, equistatin also strongly inhibits cathepsin D with Ki = 0.3 nM but not other aspartic proteinases such as pepsin, chymosin, and HIV-PR. This activity resides on the eq d-2,3 domains (Ki = 0.4 nM). Papain and cathepsin D can be bound and inhibited simultaneously by equistatin at pH 4.5, confirming the physical separation of the two binding sites. Equistatin is the first inhibitor of animal origin known to inhibit cathepsin D. The obtained results demonstrate that the widely distributed thyroglobulin type-1 domains can support a variety of functions. PMID- 9872989 TI - Human homologs of Schizosaccharomyces pombe rad1, hus1, and rad9 form a DNA damage-responsive protein complex. AB - DNA damage activates cell cycle checkpoints in yeast and human cells. In the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe checkpoint deficient mutants have been characterized, and the corresponding genes have been cloned. Searches for human homologs of S. pombe rad1, rad9, and hus1 genes identified the potential human homologs hRad1, hRad9, and hHus1; however, little is known about the roles of these proteins in human cells. The present studies demonstrate that hRad1 and hHus1 associate in a complex that interacts with a highly modified form of hRad9, but hHus1 and hRad1 do not associate with hRad17. In addition to being a key participant in complex formation, hRad9 is phosphorylated in response to DNA damage. Together, these results suggest that hRad9, hRad1, and hHus1 are central components of a DNA damage-responsive protein complex in human cells. PMID- 9872990 TI - Activation of a CrkL-stat5 signaling complex by type I interferons. AB - Type I interferons (IFNalpha and IFNbeta) transduce signals by inducing tyrosine phosphorylation of Jaks and Stats, as well as the CrkL adapter, an SH2/SH3 containing protein which provides a link to downstream pathways that mediate growth inhibition. We report that Stat5 interacts constitutively with the IFN receptor-associated Tyk-2 kinase, and during IFNalpha stimulation its tyrosine phosphorylated form acts as a docking site for the SH2 domain of CrkL. CrkL and Stat5 then form a complex that translocates to the nucleus. This IFN-inducible CrkL-Stat5 complex binds in vitro to the TTCTAGGAA palindromic element found in the promoters of a subset of IFN-stimulated genes. Thus, during activation of the Type I IFN receptor, CrkL functions as a nuclear adapter protein and, in association with Stat5, regulates gene transcription through DNA binding. PMID- 9872991 TI - Domain-specific interactions between the p185(neu) and epidermal growth factor receptor kinases determine differential signaling outcomes. AB - We expressed the epidermal growth factor receptor (EGFR) along with mutant p185(neu) proteins containing the rat transmembrane point mutation. The work concerned the study of the contributions made by various p185(neu) subdomains to signaling induced by a heterodimeric ErbB complex. Co-expression of full-length EGFR and oncogenic p185(neu) receptors resulted in an increased EGF-induced phosphotyrosine content of p185(neu), increased cell proliferation to limiting concentrations of EGF, and increases in both EGF-induced MAPK and phosphatidylinositol 3-kinase (PI 3-kinase) activation. Intracellular domain deleted p185(neu) receptors (T691stop neu) were able to associate with full length EGFR, but induced antagonistic effects on EGF-dependent EGF receptor down regulation, cell proliferation, and activation of MAPK and PI 3-kinase pathways. Ectodomain-deleted p185(neu) proteins (TDelta5) were unable to physically associate with EGFR, and extracellular domain-deleted p185(neu) forms failed to augment activation of MAPK and PI 3-kinase in response to EGF. Association of EGFR with a carboxyl-terminally truncated p185(neu) mutant (TAPstop) form did not increase transforming efficiency and phosphotyrosine content of the TAPstop species, and proliferation of EGFR.TAPstop-co-expressing cells in response to EGF was similar to cells containing EGFR only. Thus, neither cooperative nor inhibitory effects were observed in cell lines co-expressing either TDelta5 or TAPstop mutant proteins. Unlike the formation of potent homodimer assemblies composed of oncogenic p185(neu), the induction of signaling from p185(neu).EGFR heteroreceptor assemblies requires the ectodomain for ligand-dependent physical association and intracellular domain contacts for efficient intermolecular kinase activation. PMID- 9872992 TI - Endoglin is an accessory protein that interacts with the signaling receptor complex of multiple members of the transforming growth factor-beta superfamily. AB - Endoglin (CD105) is a transmembrane glycoprotein that binds transforming growth factor (TGF)-beta1 and -beta3, and coprecipitates with the Ser/Thr kinase signaling receptor complex by affinity labeling of endothelial and leukemic cells. The present study shows that in addition to TGF-beta1 and -beta3, endoglin interacts with activin-A, bone morphogenetic protein (BMP)-7, and BMP-2 but requires coexpression of the respective ligand binding kinase receptor for this association. Endoglin cannot bind ligands on its own and does not alter binding to the kinase receptors. It binds TGF-beta1 and -beta3 by associating with the TGF-beta type II receptor and interacts with activin-A and BMP-7 via activin type II receptors, ActRII and ActRIIB, regardless of which type I receptor partner is coexpressed. However, endoglin binds BMP-2 by interacting with the ligand binding type I receptors, ALK3 and ALK6. The formation of heteromeric signaling complexes was not altered by the presence of endoglin, although it was coprecipitated with these complexes. Endoglin did not interact with BMP-7 through complexes containing the BMP type II receptor, demonstrating specificity of its action. Our data suggest that endoglin is an accessory protein of multiple kinase receptor complexes of the TGF-beta superfamily. PMID- 9872993 TI - Mxi1 is a repressor of the c-Myc promoter and reverses activation by USF. AB - The basic region/helix-loop-helix/leucine zipper (B-HLH-LZ) oncoprotein c-Myc is abundant in proliferating cells and forms heterodimers with Max protein that bind to E-box sites in DNA and stimulate genes required for proliferation. A second B HLH-LZ protein, Mxi1, is induced during terminal differentiation, and forms heterodimers with Max that also bind E-boxes but tether the mSin3 transcriptional repressor protein along with histone deacetylase thereby antagonizing Myc dependent activation. We show that Mxi1 also antagonizes Myc by a second pathway, repression of transcription from the major c-myc promoter, P2. Repression was independent of Mxi1 binding to mSin3 but dependent on the Mxi1 LZ and COOH terminal sequences, including putative casein kinase II phosphorylation sites. Repression targeted elements of the myc P2 promoter core (-35/+10), where it reversed transactivation by the constitutive transcription factor, USF. We show that Zn2+ induction of a stably transfected, metallothionein promoter-regulated mxi1 gene blocked the ability of serum to induce transcription of the endogenous c-myc gene and cell entry into S phase. Thus, induction of Mxi1 in terminally differentiating cells may block Myc function by repressing the c-myc gene P2 promoter, as well as by antagonizing Myc-dependent transactivation through E boxes. PMID- 9872994 TI - The role of Tec protein-tyrosine kinase in T cell signaling. AB - The Tec protein-tyrosine kinase family includes Btk, Itk/Tsk/Emt, Tec, Rlk/Txk, and Bmx which are involved in signals mediated by various cytokines or antigen receptors. Itk is expressed primarily in T cells and activated by TCR/CD3, CD28, and CD2. However, the defect in T cell signaling in itk-deficient mice is very modest. Thus, we looked for other Tec family kinases that could be expressed in lymphoid cells and involved in T cell signal transduction. Here, we demonstrate that Tec, expressed in T cells, is activated following TCR/CD3 or CD28 ligation and interacts with CD28 receptor in an activation-dependent manner. This interaction involves the Tec SH3 domain and the proline-rich motifs in CD28. We also show that Tec can phosphorylate p62(dok), one CD28-specific substrate, whereas Itk cannot. Overexpression of Tec but not Itk can enhance the interleukin 2 promoter activity mediated by TCR/CD3 or CD28 stimulation and introduction of a kinase-dead Tec but not Itk can suppress interleukin-2 expression, indicating that Tec is directly involved in T cell activation. Altogether, these data demonstrate that Tec kinase is an integral component of T cell signaling and that the two Tec family kinases, Tec and Itk, have distinct roles in T cell activation. PMID- 9872995 TI - Recombinant pinoresinol-lariciresinol reductases from western red cedar (Thuja plicata) catalyze opposite enantiospecific conversions. AB - Although the heartwood of woody plants represents the main source of fiber and solid wood products, essentially nothing is known about how the biological processes leading to its formation are initiated and regulated. Accordingly, a reverse transcription-polymerase chain reaction-guided cloning strategy was employed to obtain genes encoding pinoresinol-lariciresinol reductases from western red cedar (Thuja plicata) as a means to initiate the study of its heartwood formation. (+)-Pinoresinol-(+)-lariciresinol reductase from Forsythia intermedia was used as a template for primer construction for reverse transcription-polymerase chain reaction amplifications, which, when followed by homologous hybridization cloning, resulted in the isolation of two distinct classes of putative pinoresinol-lariciresinol reductase cDNA clones from western red cedar. A representative of each class was expressed as a fusion protein with beta-galactosidase and assayed for enzymatic activity. Using both deuterated and radiolabeled (+/-)-pinoresinols as substrates, it was established that each class of cDNA encoded a pinoresinol-lariciresinol reductase of different (opposite) enantiospecificity. Significantly, the protein from one class converted (+) pinoresinol into (-)-secoisolariciresinol, whereas the other utilized the opposite (-)-enantiomer to give the corresponding (+)-form. This differential substrate specificity raises important questions about the role of each of these individual reductases in heartwood formation, such as whether they are expressed in different cells/tissues or at different stages during heartwood development. PMID- 9872996 TI - Evidence for a beta2-adrenergic/arachidonic acid pathway in ventricular cardiomyocytes. Regulation by the beta1-adrenergic/camp pathway. AB - The signaling pathway mediating the contractile effect of beta2-adrenergic receptors (beta2-AR) in the heart is still matter of debate. By using embryonic chick ventricular cardiomyocytes that express both functional beta1-and beta2 ARs, we show here that the specific beta2-AR agonist, zinterol, increases the amplitude of Ca2+ transients and cell contraction of electrically stimulated cells. Zinterol, up to 10 microM, did not stimulate adenylyl cyclase activity, and its effect on Ca2+ transients was unmodified by the specific cAMP antagonist, (Rp)-cAMPS. In contrast, zinterol (10-100 nM) triggered arachidonic acid (AA) release from [3H]AA-loaded cells via the activation of the cytosolic phospholipase A2 (cPLA2). Stimulation of the Ca2+ transients by zinterol was abolished by the cPLA2 inhibitor, AACOCF3, and was mimicked by AA (0.3-3 microM). Both stimulations of [3H]AA release and of [Ca2+]i cycling by zinterol were abolished after treatment of the cardiomyocytes with pertussis toxin. Although cell responses to beta2-AR stimulation were mediated by AA, they were under cAMP control as follows: (i) the beta1-AR stimulation exerted a cAMP-mediated negative constraint on the beta2-AR/cPLA2 pathway; (ii) cAMP potentiated AA action downstream beta-AR stimulation. We conclude that, in cardiomyocytes, beta2-AR is coupled to cPLA2 activation via a pertussis toxin-sensitive G protein. These results demonstrate the involvement of the cPLA2/AA pathway in mediating positive inotropic effects, which could potentially compensate for a defective cAMP pathway. PMID- 9872998 TI - DNA-interacting proteins in the spermiogenesis of the mollusc Murex brandaris. AB - Sperm chromatin of Murex brandaris (a neogastropod mollusc) undergoes a series of structural transitions during spermiogenesis. The DNA-interacting proteins responsible for these changes as well as the mature protamines present in the ripe sperm nucleus have been characterized. The results reveal that spermiogenic nuclear proteins are protamine precursors that are subjected to a substantial number of small N-terminal deletions that gradually modify their overall charge. The composition of mature protamines is remarkably simple in turn, promoting an efficient and extremely tight packaging of DNA. The pattern of spermiogenic chromatin condensation in M. brandaris clearly departs from that corresponding to vertebrate chromatin. PMID- 9872997 TI - Cellular protection mechanisms against extracellular heme. heme-hemopexin, but not free heme, activates the N-terminal c-jun kinase. AB - Hemopexin protects cells lacking hemopexin receptors by tightly binding heme abrogating its deleterious effects and preventing nonspecific heme uptake, whereas cells with hemopexin receptors undergo a series of cellular events upon encountering heme-hemopexin. The biochemical responses to heme-hemopexin depend on its extracellular concentration and range from stimulation of cell growth at low levels to cell survival at otherwise toxic levels of heme. High (2-10 microM) but not low (0.01-1 microM) concentrations of heme-hemopexin increase, albeit transiently, the protein carbonyl content of mouse hepatoma (Hepa) cells. This is due to events associated with heme transport since cobalt-protoporphyrin IX hemopexin, which binds to the receptor and activates signaling pathways without tetrapyrrole transport, does not increase carbonyl content. The N-terminal c-Jun kinase (JNK) is rapidly activated by 2-10 microM heme-hemopexin, yet the increased intracellular heme levels are neither toxic nor apoptotic. After 24 h exposure to 10 microM heme-hemopexin, Hepa cells become refractory to the growth stimulation seen with 0.1-0.75 microM heme-hemopexin but HO-1 remains responsive to induction by heme-hemopexin. Since free heme does not induce JNK, the signaling events, like phosphorylation of c-Jun via activation of JNK as well as the nuclear translocation of NFkappaB, G2/M arrest, and increased expression of p53 and of the cell cycle inhibitor p21(WAF1/CIP1/SDI1) generated by heme hemopexin appear to be of paramount importance in cellular protection by heme hemopexin. PMID- 9872999 TI - Single cell analysis and selection of living retrovirus vector-corrected mucopolysaccharidosis VII cells using a fluorescence-activated cell sorting-based assay for mammalian beta-glucuronidase enzymatic activity. AB - Mutations in the acid beta-glucuronidase gene lead to systemic accumulation of undegraded glycosaminoglycans in lysosomes and ultimately to clinical manifestations of mucopolysaccharidosis VII (Sly disease). Gene transfer by retrovirus vectors into murine mucopolysaccharidosis VII hematopoietic stem cells or fibroblasts ameliorates glycosaminoglycan accumulation in some affected tissues. The efficacy of gene therapy for mucopolysaccharidosis VII depends on the levels of beta-glucuronidase secreted by gene-corrected cells; therefore, enrichment of transduced cells expressing high levels of enzyme prior to transplantation is desirable. We describe the development of a fluorescence activated cell sorter-based assay for the quantitative analysis of beta glucuronidase activity in viable cells. Murine mucopolysaccharidosis VII cells transduced with a beta-glucuronidase retroviral vector can be isolated by cell sorting on the basis of beta-glucuronidase activity and cultured for further use. In vitro analysis revealed that sorted cells have elevated levels of beta glucuronidase activity and secrete higher levels of cross-correcting enzyme than the population from which they were sorted. Transduced fibroblasts stably expressing beta-glucuronidase after subcutaneous passage in the mucopolysaccharidosis VII mouse can be isolated by cell sorting and expanded ex vivo. A relatively high percentage of these cells maintain stable expression after secondary transplantation, yielding significantly higher levels of enzymatic activity than that generated in the primary transplant. PMID- 9873000 TI - Quantitative assessment of EF-1alpha.GTP binding to aminoacyl-tRNAs, aminoacyl viral RNA, and tRNA shows close correspondence to the RNA binding properties of EF-Tu. AB - A ribonuclease protection assay was used to determine the equilibrium dissociation constants (Kd) for the binding of various RNAs by wheat germ EF 1alpha.GTP. Aminoacylated fully modified tRNAs and unmodified tRNA transcripts of four specificities (valyl, methionyl, alanyl, and phenylalanyl) from higher plants or Escherichia coli were bound with Kd values between 0.8 and 10 nM. A valylated 3'-fragment of turnip yellow mosaic virus RNA, which has a pseudoknotted amino acid acceptor stem, was bound with affinity similar to that of Val-tRNAVal. Uncharged tRNA and initiator Met-tRNAMet from wheat germ, RNAs that are normally excluded from the ribosomal A site in vivo, bound weakly. The discrimination against wheat germ initiator Met-tRNAMet was almost entirely due to the 2'-phosphoribosyl modification at nucleotide G64, since removal resulted in tight binding by EF-1alpha.GTP. A 44-nucleotide RNA representing a kinked acceptor/T arm obtained by in vitro selection to bacterial EF-Tu formed an Ala RNA.EF-1alpha.GTP complex with a Kd of 29 nM, indicating that much of the binding affinity for aminoacylated tRNA is derived from interaction with the acceptor/T half of the molecule. The pattern of tRNA interaction observed for EF-1alpha (eEF1A) therefore closely resembles that of bacterial EF-Tu (EF1A). PMID- 9873001 TI - Mutational analysis of aspartate residues in the transmembrane regions and cytoplasmic loops of rat vesicular acetylcholine transporter. AB - The vesicular acetylcholine transporter (VAChT) is responsible for the transport of the neurotransmitter acetylcholine (ACh) into synaptic vesicles using an electrochemical gradient to drive transport. Rat VAChT has a number of aspartate residues within its predicted transmembrane domains (TM) and cytoplasmic loops, which may play important structural or functional roles in acetylcholine transport. In order to identify functional charged residues, site-directed mutagenesis of rVAChT was undertaken. No effect on ACh transport was observed when any of the five aspartate residues in the cytoplasmic loop were converted to asparagine. Similarly, changing Asp-46 (D46N) in TM1 or Asp-255 (D255N) in TM6 had no effect on ACh transport or vesamicol binding. However, replacement of Asp 398 in TM10 with Asn completely eliminated both ACh transport and vesamicol binding. The conservative mutant D398E retained transport activity, but not vesamicol binding, suggesting this residue is critical for transport. Mutation of Asp-193 in TM4 did not affect ACh transport activity; however, vesamicol binding was dramatically reduced. With mutant D425N of TM11 transport activity for ACh was completely blocked, without an effect on vesamicol binding. Activity was not restored in the conservative mutant D425E, suggesting the side chain as well as the negative charge of Asp-425 is important for substrate binding. These mutants, as well as mutant D193N, clearly dissociated ACh binding and transport from vesamicol binding. These data suggest that Asp-398 in TM10 and Asp-425 in TM11 are important for ACh binding and transport, while Asp-193 and Asp-398 in TM4 and TM10, respectively, are involved in vesamicol binding. PMID- 9873002 TI - Interaction of the sarcin/ricin domain of 23 S ribosomal RNA with proteins L3 and L6. AB - We investigated interaction of an RNA domain covering the target site of alpha sarcin and ricin (sarcin/ricin domain) of Escherichia coli 23 S rRNA with ribosomal proteins. RNA fragments comprising residues 2630-2788 (Tox-1) and residues 2640-2774 (Tox-2) of 23 S rRNA were transcribed in vitro and used to analyze the binding proteins by gel shift and filter binding. Protein L6 bound to both Tox-1 (Kd: 0.31 microM) and Tox-2 (Kd: 0.18 microM), and L3 bound only to Tox-1 (Kd: 0.069 microM) in a solution containing 10 mM MgCl2 and 175 mM KCl at 0 degreesC. Footprinting studies were performed using the chemical probe dimethyl sulfate on full-length 23 S rRNA. Binding of L6 protected a single base, A-2757, and strongly enhanced reactivity of C-2752. A direct role of A-2757 in the L6 binding was verified by site-directed mutagenesis; replacements of A-2757 with G and C impaired the L6 binding. On the other hand, binding of L3 protected A-2632, A-2634, A-2635, A-2675, A-2726, A-2733, A-2749, and A-2750. Interestingly, binding of L6 and L3 together protected additional bases A-2657, A-2662, C-2666, and C-2667 in the sarcin/ricin loop, in addition to A-2740, A-2741, A-2748, A 2753, A-2764, A-2765, and A-2766 in the other stem-loop. This appears to be due to cooperative interaction of L3 and L6 with the RNA. The results are discussed with respect to conformational modulation of the sarcin/ricin domain by the protein binding. PMID- 9873003 TI - Identification of kinase-phosphatase signaling modules composed of p70 S6 kinase protein phosphatase 2A (PP2A) and p21-activated kinase-PP2A. AB - A growing body of evidence indicates that regulation of protein-serine/threonine phosphatase 2A (PP2A) involves its association with other cellular and viral proteins in multiprotein complexes. PP2A-containing protein complexes may exist that contribute to PP2A's important regulatory role in many cellular processes. To identify such protein complexes, PP2A was partially purified from rat brain soluble extracts following treatment with a reversible cross-linker to stabilize large molecular size forms of PP2A. Compared with native (uncross-linked) PP2A, cross-linked PP2A revealed an enrichment of p70 S6 kinase and two p21-activated kinases (PAK1 and PAK3) in the PP2A complex, indicating these kinases may associate with PP2A. The existence of protein kinase-PP2A complexes in rat brain soluble extracts was further substantiated by the following results: 1) independent immunoprecipitation of the kinases revealed that PP2A co-precipitated with p70 S6 kinase and the two PAK isoforms; 2) glutathione S-transferase fusion proteins of p70 S6 kinase and PAK3 each isolated PP2A; and 3) PAK3 and p70 S6 kinase bound to microcystin-Sepharose (an affinity resin for PP2A-PP1). Cumulatively, these findings provide evidence for association of PP2A with p70 S6 kinase, PAK1, and PAK3 in the context of the cellular environment. Moreover, together with the recent reports describing associations of PP2A with Ca2+/calmodulin-dependent protein kinase IV (Westphal, R. S., Anderson, K. A., Means, A. R., and Wadzinski, B. E. (1998) Science 280, 1258-1261) and casein kinase IIalpha (Heriche, J. K., Lebrin, F., Rabilloud, T., Leroy, D., Chambaz, E. M., and Goldberg, Y. (1997) Science 276, 952-955), the present data provide compelling evidence for the existence of protein kinase-PP2A signaling modules as a new paradigm for the control of various intracellular signaling cascades. PMID- 9873004 TI - Measurement of resting cytosolic Ca2+ concentrations and Ca2+ store size in HEK 293 cells transfected with malignant hyperthermia or central core disease mutant Ca2+ release channels. AB - Malignant hyperthermia (MH) and central core disease (CCD) mutations were introduced into full-length rabbit Ca2+ release channel (RYR1) cDNA, which was then expressed transiently in HEK-293 cells. Resting Ca2+ concentrations were higher in HEK-293 cells expressing homotetrameric CCD mutant RyR1 than in cells expressing homotetrameric MH mutant RyR1. Cells expressing homotetrameric CCD or MH mutant RyR1 exhibited lower maximal peak amplitudes of caffeine-induced Ca2+ release than cells expressing wild type RyR1, suggesting that MH and CCD mutants might be "leaky." In cells expressing homotetrameric wild type or mutant RyR1, the amplitude of 10 mM caffeine-induced Ca2+ release was correlated significantly with the amplitude of carbachol- or thapsigargin-induced Ca2+ release, indicating that maximal drug-induced Ca2+ release depends on the size of the endoplasmic reticulum Ca2+ store. The content of endogenous sarco(endo)plasmic reticulum Ca2+ ATPase isoform 2b (SERCA2b), measured by enzyme-linked immunosorbent assay, 45Ca2+ uptake, and confocal microscopy, was increased in HEK-293 cells expressing wild type or mutant RyR1, supporting the view that endoplasmic reticulum Ca2+ storage capacity is increased as a compensatory response to an enhanced Ca2+ leak. When heterotetrameric (1:1) combinations of MH/CCD mutant and wild type RyR1 were expressed together with SERCA1 to enhance Ca2+ reuptake, the amplitude of Ca2+ release in response to low concentrations of caffeine and halothane was higher than that observed in cells expressing wild type RyR1 and SERCA1. In Ca2+ free medium, MH/CCD mutants were more sensitive to caffeine than wild type RyR1, indicating that caffeine hypersensitivity observed with a variety of MH/CCD mutant RyR1 proteins is not dependent on extracellular Ca2+ concentration. PMID- 9873005 TI - Alternatively spliced variant of Smad2 lacking exon 3. Comparison with wild-type Smad2 and Smad3. AB - An alternatively spliced variant of Smad2 with a deletion of exon 3 (Smad2Deltaexon3) is found in various cell types. Here, we studied the function of Smad2Deltaexon3 and compared it with those of wild-type Smad2 containing exon 3 (Smad2(wt)) and Smad3. When transcriptional activity was measured using the p3TP-lux construct, Smad2Deltaexon3 was more potent than Smad2(wt), and had activity similar to Smad3. Transcriptional activation of the activin-responsive element (ARE) of Mix.2 gene promoter by Smad2Deltaexon3 was also similar to that by Smad3, and slightly less potent than that by Smad2(wt). Phosphorylation by the activated transforming growth factor-beta type I receptor and heteromer formation with Smad4 occurred to similar extents in Smad2Deltaexon3, Smad2(wt), and Smad3. However, DNA binding to the activating protein-1 sites of p3TP-lux was observed in Smad2Deltaexon3 as well as in Smad3, but not in Smad2(wt). In contrast, Smad2(wt), Smad2Deltaexon3, and Smad3 efficiently formed ARE-binding complexes with Smad4 and FAST1, although Smad2(wt) did not directly bind to ARE. These results suggest that exon 3 of Smad2 interferes with the direct DNA binding of Smad2, and modifies the function of Smad2 in transcription of certain target genes. PMID- 9873006 TI - High level transcription of the complement regulatory protein CD59 requires an enhancer located in intron 1. AB - CD59 is a complement regulatory protein and may also act as a signal-transducing molecule. CD59 transgenic mice have been generated using a CD59 minigene (CD59 minigene-1). Although this minigene contained a 4.6-kilobase pair 5'-flanking region from the human CD59 gene as a promoter, the expression levels of the CD59 mRNA were substantially lower than those observed in humans, suggesting that CD59 gene expression might also require other transcriptional regulatory elements such as an enhancer. To investigate the transcriptional regulation of the CD59 gene, we used three cell lines that express CD59 at different levels. We have identified DNase I-hypersensitive sites in intron 1 in HeLa cells, which express CD59 at high levels, but not in Jurkat (intermediate level) or Raji cells (low level). Furthermore, cell line-specific enhancer activity was detected in a fragment containing these DNase I-hypersensitive sites. The CD59 enhancer was mapped to between -1155 and -888 upstream of the 5'-end of exon 2. To investigate the enhancer activity in vivo, a new CD59 minigene was constructed by the addition of the enhancer fragment into CD59 minigene-1. High expressor CD59 transgenic mice were generated using the new minigene. PMID- 9873007 TI - Domain organization of the 39-kDa receptor-associated protein. AB - The 39-kDa receptor-associated protein (RAP) is an endoplasmic reticulum resident protein that binds to the low density lipoprotein receptor-related protein (LRP) as well as certain members of the low density lipoprotein receptor superfamily and antagonizes ligand binding. In order to identify important functional regions of RAP, studies were performed to define the domain organization and domain boundaries of this molecule. Differential scanning calorimetry (DSC) experiments revealed that the process of thermal denaturation of RAP is highly reversible and occurs in a broad temperature range with two well resolved heat absorption peaks. A good fit of the endotherm was obtained with four two-state transitions suggesting these many cooperative domains in the molecule. A number of recombinant fragments of RAP were expressed in bacteria, and their domain composition and stability were characterized by DSC, circular dichroism, and fluorescence spectroscopy. The results confirmed that RAP is composed of four independently folded domains, D1, D2, D3, and D4, that encompass residues 1-92, 93-163, 164-216, and 217-323, respectively. The first and the fourth domains preserved their structure and stability when isolated, whereas the compact structure of the fragment corresponding to D2 seems to be altered when isolated from the parent molecule. Isolated D3 was partially degraded during isolation from bacterial lysates. The isolated D4 was capable of binding with high affinity to LRP whereas neither D1 nor D2 bound. At the same time a fragment containing both D1 and D2 exhibited high affinity binding to LRP. These facts combined with the thermodynamic analysis of the melting process of the fragments containing D1 and D2 indicate that these two domains interact with each other and that the proper folding of the second domain into a native-like active conformation requires presence of the first domain. PMID- 9873008 TI - Critical factors in basal cell adhesion molecule/lutheran-mediated adhesion to laminin. AB - Basal cell adhesion molecule (B-CAM) and Lutheran (LU) are two spliceoforms of a single immunoglobulin superfamily protein containing five Ig domains and comprise the sickle (SS) red cell receptor for laminin. We have now analyzed laminin binding to murine erythroleukemia cells transfected with various human B-CAM/LU constructs. B-CAM and LU bound equally well to laminin, indicating that the longer cytoplasmic tail of LU is not required for binding. However, binding of soluble laminin did require the presence of the membrane-proximal fifth immunoglobulin superfamily (IgSF) domain of LU, while deletion of IgSF domains 1, 2, 3, or 4 individually or together did not abrogate laminin binding. Under flow conditions, MEL cells expressing B-CAM, LU, and LU lacking domains 1, 2, 3, or 4 adhered to immobilized laminin with critical shear stresses over 10 dynes/cm2. However, MEL cells expressing LU lacking domain 5 bound to laminin poorly (critical shear stress = 2.3 dynes/cm2). Moreover, expression of only IgSF domain 5 of LU was sufficient to mediate MEL cell adhesion to immobilized laminin (critical shear stress >10 dynes/cm2). Finally, Scatchard analysis showed that SS red cells had an average of 67% more B-CAM/LU than normal red cells, and low density red cells from sickle cell disease patients expressed 40-55% more B CAM/LU than high density SS red cells. B-CAM/LU copy number thus may also play a role in the abnormal adhesion of SS red cells to laminin. PMID- 9873009 TI - Phospholipase D mediates matrix metalloproteinase-9 secretion in phorbol ester stimulated human fibrosarcoma cells. AB - Phospholipase D (PLD) has been implicated in vesicle trafficking in the Golgi and hence secretion. In this study, we show that the secretion of matrix metalloproteinase-9 (MMP-9) from HT 1080 human fibrosarcoma cells was stimulated by phorbol 12-myristate 13-acetate in a time- and dose-dependent manner that involved protein kinase C. The phorbol ester also increased PLD activity in the cells. Evidence that PLD was involved in the stimulation of MMP-9 secretion was provided by the observations that the secretion of MMP-9 was stimulated by the introduction of short-chain phosphatidic acid (PA) into the growth medium and that inhibition of PA production by 1-propanol inhibited secretion. Using a short chain diacylglycerol we excluded the possibility that MMP-9 secretion was induced by diacylglycerol formed from PA by phosphatidic acid phosphatase. Furthermore, propranolol, an inhibitor of this enzyme, had no effect on secretion induced by either phorbol 12-myristate 13-acetate or PA. The data presented here indicate that activation of protein kinase C increases MMP-9 secretion in HT 1080 cells and implicate PLD and PA formation in the effect. PMID- 9873010 TI - Both lobes of the soluble receptor of the periplasmic histidine permease, an ABC transporter (traffic ATPase), interact with the membrane-bound complex. Effect of different ligands and consequences for the mechanism of action. AB - The histidine permease of Salmonella typhimurium is an ABC transporter (traffic ATPase). The liganded soluble receptor, the histidine-binding protein HisJ, interacts with the membrane-bound complex HisQMP2 and stimulates its ATPase activity, which results in histidine translocation. In this study, we utilized HisJ proteins with mutations in either of the two lobes and wild type HisJ liganded with different substrates to show that each lobe carries an interaction site and that both lobes are involved in inducing (stimulating) the ATPase activity. We suggest that the spatial relationship between the lobes is one of the factors recognized by the membrane-bound complex in dictating the efficiency of the induction signal and of translocation. Several of the key residues involved have been identified. In addition, using constitutive ATPase mutants, we show that the binding protein provides some additional essential function(s) in translocation that is independent of the stimulation of ATP hydrolysis, and one possible mechanism is proposed, which includes the notion that liganded HisJ has different optimal conformations for signaling and for translocation. PMID- 9873011 TI - The flavohemoglobin of Escherichia coli confers resistance to a nitrosating agent, a "Nitric oxide Releaser," and paraquat and is essential for transcriptional responses to oxidative stress. AB - Escherichia coli possesses a flavohemoglobin (Hmp), product of hmp, the first microbial globin gene to be sequenced and characterized at the molecular level. Although related proteins occur in numerous prokaryotes and eukaryotic microorganisms, the function(s) of these proteins have been elusive. Here we report construction of a defined hmp mutation and its use to probe Hmp function. As anticipated from up-regulation of hmp expression by nitric oxide (NO), S nitrosoglutathione (GSNO) or sodium nitroprusside (SNP), the hmp mutant is hypersensitive to these agents. The hmp promoter is more sensitive to SNP and S nitroso-N-penicillamine (SNAP) than is the soxS promoter, consistent with the role of Hmp in protection from reactive nitrogen species. Additional functions for Hmp are indicated by (a) parallel sensitivity of the hmp mutant to the redox cycling agent, paraquat, (b) inability of the mutant to up-regulate fully the soxS and sodA promoters in response to oxidative stress caused by paraquat, GSNO and SNP, and (c) failure of the mutant to accumulate reduced paraquat radical after anoxic growth. We conclude that Hmp plays a role in protection from nitrosating agents and NO-related species and oxidative stress. This protective role probably involves direct detoxification of those species and sensing of NO related and oxidative stress. PMID- 9873012 TI - Purinergic inhibition of glucose transport in cardiomyocytes. AB - ATP is known to act as an extracellular signal in many organs. In the heart, extracellular ATP modulates ionic processes and contractile function. This study describes a novel, metabolic effect of exogenous ATP in isolated rat cardiomyocytes. In these quiescent (i.e. noncontracting) cells, micromolar concentrations of ATP depressed the rate of basal, catecholamine-stimulated, or insulin-stimulated glucose transport by up to 60% (IC50 for inhibition of insulin dependent glucose transport, 4 microM). ATP decreased the amount of glucose transporters (GLUT1 and GLUT4) in the plasma membrane, with a concomitant increase in intracellular microsomal membranes. A similar glucose transport inhibition was produced by P2 purinergic agonists with the following rank of potencies: ATP approximately ATPgammaS approximately 2-methylthio-ATP (P2Y selective) > ADP > alpha,betameATP (P2X-selective), whereas the P1 purinoceptor agonist adenosine was ineffective. The effect of ATP was suppressed by the poorly subtype-selective P2 antagonist pyridoxal-phosphate-6-azophenyl-2', 4'-disulfonic acid but, surprisingly, not by the nonselective antagonist suramin nor by the P2Y specific Reactive Blue 2. Glucose transport inhibition by ATP was not affected by a drastic reduction of the extracellular concentrations of calcium (down to 10( 9) M) or sodium (down to 0 mM), and it was not mimicked by a potassium-induced depolarization, indicating that purinoceptors of the P2X family (which are nonselective cation channels whose activation leads to a depolarizing sodium and calcium influx) are not involved. Inhibition was specific for the transmembrane transport of glucose because ATP did not inhibit (i) the rate of glycolysis under conditions where the transport step is no longer rate-limiting nor (ii) the rate of [1-14C]pyruvate decarboxylation. In conclusion, extracellular ATP markedly inhibits glucose transport in rat cardiomyocytes by promoting a redistribution of glucose transporters from the cell surface to an intracellular compartment. This effect of ATP is mediated by P2 purinoceptors, possibly by a yet unknown subtype of the P2Y purinoceptor family. PMID- 9873013 TI - Expression, purification, and spectroscopic characterization of human thromboxane synthase. AB - Thromboxane A2 (TXA2) is a potent inducer of vasoconstriction and platelet aggregation. Large scale expression of TXA2 synthase (TXAS) is very useful for studies of the reaction mechanism, structural/functional relationships, and drug interactions. We report here a heterologous system for overexpression of human TXAS. The TXAS cDNA was modified by replacing the sequence encoding the first 28 amino acid residues with a CYP17 amino-terminal sequence and by adding a polyhistidine tag sequence prior to the stop codon; the cDNA was inserted into the pCW vector and co-expressed with chaperonins groES and groEL in Escherichia coli. The resulting recombinant protein was purified to electrophoretic homogeneity by affinity, ion exchange, and hydrophobic chromatography. UV-visible absorbance (UV-Vis), magnetic circular dichroism (MCD), and electron paramagnetic resonance (EPR) spectra indicate that TXAS has a typical low spin cytochrome P450 heme with an oxygen-based distal ligand. The UV-Vis and EPR spectra of recombinant TXAS were essentially identical to those of TXAS isolated from human platelets, except that a more homogenous EPR spectrum was observed for the recombinant TXAS. The recombinant protein had a heme:protein molar ratio of 0.7:1 and a specific activity of 12 micromol of TXA2/min/mg of protein at 23 degreesC. Furthermore, it catalyzed formation of TXA2, 12-hydroxy-5,8,10-heptadecatrienoic acid, and malondialdehyde in a molar ratio of 0.94:1.0:0.93. Spectral binding titrations showed that bulky heme ligands such as clotrimazole bound strongly to TXAS (Kd approximately 0.5 microM), indicating ample space at the distal face of the heme iron. Analysis of MCD and EPR spectra showed that TXAS was a typical low spin hemoprotein with a proximal thiolate ligand and had a very hydrophobic distal ligand binding domain. PMID- 9873014 TI - Regulation by pH of the alternative splicing of the stem cell factor pre-mRNA in the testis. AB - Proliferation and differentiation of progenitor stem cells are mainly controlled by diffusible and adhesion molecules. Stem cell factor (SCF), an essential regulator of spermatogenesis produced by Sertoli cells, utilize both modes of cell to cell communication. Indeed, SCF exists in soluble (SCFs) and membrane bound (SCFm) forms, which are required for a complete spermatogenesis, and are generated by alternative splicing of optional exon 6, encoding sites of proteolysis. We show that in the mouse testis, the alternative splicing of SCF is developmentally regulated. SCFs predominates in fetal and neonatal gonads and is then replaced by SCFm in the prepubertal and adult gonads. By sequencing SCF exon 6, we show that the flanking intronic sequences perfectly follow the gt-at rule, suggesting that the basal splicing machinery might not be responsible by itself for exon 6 skipping. Moreover, freshly isolated Sertoli cells mainly express SCFm, but a switch to SCFs occurs after 48 h of culture. We found that this change can be prevented by acidification of the culture medium at pH 6.3 or by addition of lactate. The sustained synthesis of SCFm at low pH was no longer observed in the presence of cycloheximide, suggesting that SCF exon 6 skipping requires de novo protein synthesis. Accordingly, UV cross-linking experiments show that nuclear Sertoli cell protein(s) bind in a sequence-specific manner to exon 6. Together, our data allow the proposal of an integrated mechanism in which the synthesis of lactate by Sertoli cells is used in the same time as an energetic substrate for germ cells and as a promoter of their survival/proliferation through the production of SCFm. PMID- 9873015 TI - Control of glycogen synthesis in cultured human muscle cells. AB - The regulation of glycogen synthesis and associated enzymes was studied in human myoblasts and myotubes maintained in culture. Both epidermal growth factor (EGF) and insulin stimulated glycogen synthesis approximately 2-fold, this stimulation being accompanied by a rapid and stable activation of the controlling enzyme glycogen synthase (GS). EGF also caused inhibition of glycogen synthase kinase 3 (GSK-3) and activation of the alpha isoform of protein kinase B (PKB) with the time-course and magnitude of its effects being similar to those induced by insulin. An inhibitor of the mitogen-activated protein (MAP) kinase pathway did not prevent stimulation of GS by EGF, suggesting that this pathway is not essential for the effect. A partial decrease in the fold activation of GS was, however, observed when p70(S6k) activation was blocked with rapamycin, suggesting a contribution of this pathway to the control of GS by either hormone. Wortmannin, a selective inhibitor of phosphatidylinositol 3'-kinase (PI-3 kinase) completely blocked the effects of both EGF and insulin in these cells. These results demonstrate that EGF, like insulin, activates glycogen synthesis in muscle, acting principally via the PKB/GSK-3 pathway but with a contribution from a rapamycin-sensitive component that lies downstream of PI-3 kinase. PMID- 9873016 TI - An evolutionarily conserved family of Hsp70/Hsc70 molecular chaperone regulators. AB - Heat Shock Protein 70 kDa (Hsp70) family molecular chaperones play critical roles in protein folding and trafficking in all eukaryotic cells. The mechanisms by which Hsp70 family chaperones are regulated, however, are only partly understood. BAG-1 binds the ATPase domains of Hsp70 and Hsc70, modulating their chaperone activity and functioning as a competitive antagonist of the co-chaperone Hip. We describe the identification of a family of BAG-1-related proteins from humans (BAG-2, BAG-3, BAG-4, BAG-5), the invertebrate Caenorhabditis elegans (BAG-1, BAG 2), and the fission yeast Schizosaccharomyces pombe (BAG-1A, BAG-1B). These proteins all contain a conserved approximately 45-amino acid region near their C termini (the BAG domain) that binds Hsc70/Hsp70, but they differ widely in their N-terminal domains. The human BAG-1, BAG-2, and BAG-3 proteins bind with high affinity (KD congruent with 1-10 nM) to the ATPase domain of Hsc70 and inhibit its chaperone activity in a Hip-repressible manner. The findings suggest opportunities for specification and diversification of Hsp70/Hsc70 chaperone functions through interactions with various BAG-family proteins. PMID- 9873018 TI - Purification and characterization of NAD:Penicillamine ADP transferase from Bacillus sphaericus. A novel NAD-dependent enzyme catalyzing phosphoramide bond formation. AB - A strain of Bacillus sphaericus isolated from a local soil sample has been found to use beta,beta-dimethyl-DL-cysteine (DL-penicillamine) as the sole nitrogen source. Crude cell extract of the bacterium showed potent penicillamine-consuming activity only in the presence of NAD, which, however, was not used as an electron acceptor. Characterization of reaction products revealed that penicillamine was derivatized to a phosphoramide adduct with the ADP moiety of NAD, whereas the nicotinamide-ribose group was released and hydrolyzed spontaneously to ribose and nicotinamide. The phosphoramide product, ADP-penicillamine, caused potent product inhibition on the purified enzyme, and adenylate deaminase was found to be effective in converting the inhibitory product into inosine-diphosphate penicillamine and thereby maintained the catalysis for several hours. The novel enzyme, termed as NAD:penicillamine ADP transferase, showed a single band on SDS polyacrylamide gel electrophoresis with a mass of approximately 42 kDa. The native enzyme was monomeric. The enzyme showed high substrate specificity to NAD (Km = 13.0 mM) and L-penicillamine (Km = 6.5 mM); other nucleotides such as NADP, NAD(P)H, AMP, ADP, and ADP-ribose did not substitute for NAD, and L-valine, L cysteine, L-homocysteine, L-cystine, L-leucine, and L-isoleucine did not serve as the substrate. Kinetic studies suggested an Ordered Bi Bi mechanism, with NAD as the first substrate to bind and ADP-L-penicillamine as the last product released. The novel NAD-dependent enzyme may catalyze the first step in penicillamine degradation in the strain of B. sphaericus. PMID- 9873017 TI - Tumor necrosis factor-alpha-inducible IkappaBalpha proteolysis mediated by cytosolic m-calpain. A mechanism parallel to the ubiquitin-proteasome pathway for nuclear factor-kappab activation. AB - The cytokine tumor necrosis factor alpha (TNF-alpha) induces expression of inflammatory gene networks by activating cytoplasmic to nuclear translocation of the nuclear factor-kappaB (NF-kappaB) transcription factor. NF-kappaB activation results from sequential phosphorylation and hydrolysis of the cytoplasmic inhibitor, IkappaBalpha, through the 26 S proteasome. Here, we show a parallel proteasome-independent pathway for cytokine-inducible IkappaBalpha proteolysis in HepG2 liver cells mediated by cytosolic calcium-activated neutral protease (calpains). Pretreatment with either calpain- or proteasome-selective inhibitors partially blocks up to 50% of TNF-alpha-inducible IkappaBalpha proteolysis; pretreatment with both is required to completely block IkappaBalpha proteolysis. Similarly, in transient cotransfection assays, expression of the specific inhibitor, calpastatin, partially blocks TNF-alpha-inducible NF-kappaB-dependent promoter activity and IkappaBalpha proteolysis. In TNF-alpha-stimulated cells, a rapid (within 1 min), 2.2-fold increase in cytosolic calpain proteolytic activity is measured using a specific fluorescent assay. Inducible calpain proteolytic activity occurs coincidentally with the particulate-to-cytosol redistribution of the catalytic m-calpain subunit into the IkappaBalpha compartment. Addition of catalytically active m-calpain into broken cells was sufficient to produce ligand independent IkappaBalpha proteolysis and NF-kappaB translocation. As additional evidence for calpain-dependent IkappaBalpha proteolysis and NF-kappaB activation, we demonstrate that this process occurs in a cell line (ts20b) deficient in the ubiquitin-proteasome pathway. Following inactivation of the temperature-sensitive ubiquitin-activating enzyme, IkappaBalpha proteolysis occurs in a manner sensitive only to calpain inhibitors. Our results demonstrate that TNF-alpha activates cytosolic calpains, a parallel pathway that degrades IkappaBalpha and activates NF-kappaB activation independently of the ubiquitin-proteasome pathway. PMID- 9873019 TI - Role of mitogen-activated protein kinases and c-Jun/AP-1 trans-activating activity in the regulation of protease mRNAs and the malignant phenotype in NIH 3T3 fibroblasts. AB - Ras activates a multitude of downstream activities with roles in cellular proliferation, invasion and metastasis, differentiation, and programmed cell death. In this work we have evaluated the requirement of extracellular signal regulated protein kinase (ERK), c-Jun NH2-terminal kinase kinase (JNKK), and c Jun/AP-1 activities in transformation and extracellular matrix invasion of ras oncogene expressing NIH 3T3 fibroblasts by expressing stable mutant genes that constitutively inhibit these activities. Whereas the inhibition of ERK activity reverts the transformed and invasive phenotype, the inhibition of the JNK pathway and AP-1 trans-activating activities by JNKK[K129R] and c-Jun(TAM67) had no effect on the ability of the ras oncogene-expressing cells to grow in soft agar or invade Matrigel basement membrane. Thus an elevated JNK activity and/or c Jun/AP-1 trans-activating activity are not absolute requirements for ras transformation or invasion through basement membrane, and the dependence on AP-1 activity for transformation is cell-specific. However, inhibition of JNK kinase (JNKK) in ras-transformed cells with normally elevated JNK activity switches the protease-dependent invasive phenotype from a urokinase plasminogen activator (uPA)-dependent to a cathepsin L (CL)-dependent invasive phenotype. Conversely, treatment of ras-transformed cells of low constitutive JNK activity with the JNK stimulator, anisomycin, converts the protease mRNA levels from those characteristic of a CL-dependent to a uPA-dependent phenotype. These protease phenotypes can be duplicated in untransformed NIH 3T3 cells that express platelet derived growth factor receptors and m1 muscarinic receptors that selectively stimulate the ERK or JNK pathways, respectively. It is concluded that high ERK activity is required for both protease phenotypes, whereas the JNK pathway and c Jun/AP-1 activity are not required for transformation but regulate a switch between uPA and CL protease phenotypes in both transformed and untransformed cells. In ras-transformed NIH 3T3 fibroblasts, the uPA- and CL-dependent protease phenotypes are redundant in their ability to invade through basement membrane. PMID- 9873021 TI - On the advantage of being a dimer, a case study using the dimeric Serratia nuclease and the monomeric nuclease from Anabaena sp. strain PCC 7120. AB - The extracellular endonucleases from Serratia marcescens and Anabaena sp. are members of a family of nonspecific endonucleases. In contrast to the monomeric Anabaena nuclease, the Serratia nuclease is a dimer of two identical subunits. To find out whether the two active sites of the Serratia nuclease function independently of each other and what the advantage of being a dimer for this enzyme might be, we produced (i) dimers in which the two subunits were cross linked, (ii) heterodimers consisting of a wild type and an inactive mutant subunit which were also cross-linked, and (iii) monomeric variants which are unable to dimerize. The monomeric H184R variant and the cross-linked S140C variant exhibit the same activity as the wild type enzyme, while the cross-linked heterodimer with one inactive subunit shows only half of the activity of the wild type enzyme, demonstrating functional independence of the two subunits of the Serratia nuclease. On the other hand at low enzyme and substrate concentrations dimeric forms of the Serratia nuclease are relatively more active than monomeric forms or the monomeric Anabaena nuclease in cleaving polynucleotides, not, however, oligonucleotides, which is correlated with the ability of dimeric forms of the Serratia nuclease to form large enzyme-substrate networks with high molecular weight DNA and to cleave polynucleotides in a processive manner. We conclude that in the natural habitat of Serratia marcescens where the supply of nutrients may become growth limiting the dimeric nuclease can fulfil its nutritive function more efficiently than a monomeric enzyme. PMID- 9873020 TI - S-Adenosylmethionine-dependent methylation in Saccharomyces cerevisiae. Identification of a novel protein arginine methyltransferase. AB - We used sequence motifs conserved in S-adenosylmethionine-dependent methyltransferases to identify 26 putative methyltransferases from the complete genome of the yeast Saccharomyces cerevisiae. Seven sequences with the best matches to the methyltransferase consensus motifs were selected for further study. We prepared yeast disruption mutants of each of the genes encoding these sequences, and we found that disruption of the YJL125c gene is lethal, whereas disruptions of YCR047c and YDR140w lead to slow growth phenotypes. Normal growth was observed when the YDL201w, YDR465c, YHR209w, and YOR240w genes were disrupted. Initial analysis of protein methylation patterns of all mutants by amino acid analysis revealed that the YDR465c mutant has a defect in the methylation of the delta-nitrogen atom of arginine residues. We propose that YDR465c codes for the methyltransferase responsible for this recently characterized type of protein methylation, and we designate the enzyme as Rmt2 (protein arginine methyltransferase). In addition, we show that the methylation of susceptible residues in Rmt2 substrates is likely to take place on nascent polypeptide chains and that these substrates exist in the cell as fully methylated species. Interestingly, Rmt2 has 27% sequence identity over 138 amino acids to the mammalian guanidinoacetate N-methyltransferase, an enzyme responsible for methylating the delta-nitrogen of the small molecule guanidinoacetate. PMID- 9873022 TI - Hydrogen peroxide induces intracellular calcium overload by activation of a non selective cation channel in an insulin-secreting cell line. AB - Fura-2 fluorescence was used to investigate the effects of H2O2 on [Ca2+]i in the insulin-secreting cell line CRI-G1. H2O2 (1-10 mM) caused a biphasic increase in free [Ca2+]i, an initial rise observed within 3 min and a second, much larger rise following a 30-min exposure. Extracellular calcium removal blocked the late, but not the initial, rise in [Ca2+]i. Thapsigargin did not affect either response to H2O2, but activated capacitive calcium entry, an action abolished by 10 microM La3+. Simultaneous recordings of membrane potential and [Ca2+]i demonstrated the same biphasic [Ca2+]i response to H2O2 and showed that the late increase in [Ca2+]i coincided temporally with cell membrane potential collapse. Buffering Ca2+i to low nanomolar levels prevented both phases of increased [Ca2+]i and the H2O2-induced depolarization. The H2O2-induced late rise in [Ca2+]i was prevented by extracellular application of 100 microM La3+. La3+ (100 microM) inhibited the H2O2-induced cation current and NAD-activated cation (NSNAD) channel activity in these cells. H2O2 increased the NAD/NADH ratio in intact CRI-G1 cells, consistent with increased cellular [NAD]. These data suggest that H2O2 increases [NAD], which, coupled with increased [Ca2+]i, activates NSNAD channels, causing unregulated Ca2+ entry and consequent cell death. PMID- 9873023 TI - Kinetics of peroxynitrite reaction with amino acids and human serum albumin. AB - An initial rate approach was used to study the reaction of peroxynitrite with human serum albumin (HSA) through stopped-flow spectrophotometry. At pH 7.4 and 37 degreesC, the second order rate constant for peroxynitrite reaction with HSA was 9.7 +/- 1.1 x 10(3) M-1 s-1. The rate constants for sulfhydryl-blocked HSA and for the single sulfhydryl were 5.9 +/- 0.3 and 3.8 +/- 0.8 x 10(3) M-1 s-1, respectively. The corresponding values for bovine serum albumin were also determined. The reactivity of sulfhydryl-blocked HSA increased at acidic pH, whereas plots of the rate constant with the sulfhydryl versus pH were bell shaped. The kinetics of peroxynitrite reaction with all free L-amino acids were determined under pseudo-first order conditions. The most reactive amino acids were cysteine, methionine, and tryptophan. Histidine, leucine, and phenylalanine (and by extension tyrosine) did not affect peroxynitrite decay rate, whereas for the remaining amino acids plots of kobs versus concentration were hyperbolic. The sum of the contributions of the constituent amino acids of the protein to HSA reactivity was comparable to the experimentally determined rate constant, where cysteine and methionine (seven residues in 585) accounted for an estimated 65% of the reactivity. Nitration of aromatic amino acids occurred in HSA following peroxynitrite reaction, with nitration of sulfhydryl-blocked HSA 2-fold higher than native HSA. Carbon dioxide accelerated peroxynitrite decomposition, enhanced aromatic amino acid nitration, and partially inhibited sulfhydryl oxidation of HSA. Nitration in the presence of carbon dioxide increased when the sulfhydryl was blocked. Thus, cysteine 34 was a preferential target of peroxynitrite both in the presence and in the absence of carbon dioxide. PMID- 9873024 TI - Kinetic analysis of tentoxin binding to chloroplast F1-ATPase. A model for the overactivation process. AB - The mechanism of action of tentoxin on the soluble part (chloroplast F1 H+ ATPase; CF1) of chloroplast ATP synthase was analyzed in the light of new kinetic and equilibrium experiments. Investigations were done regarding the functional state of the enzyme (activation, bound nucleotide, catalytic turnover). Dialysis and binding data, obtained with 14C-tentoxin, fully confirmed the existence of two tentoxin binding sites of distinct dissociation constants consistent with the observed Kinhibition and Koveractivation. This strongly supports a two-site model of tentoxin action on CF1. Kinetic and thermodynamic parameters of tentoxin binding to the first site (Ki = 10 nM; kon = 4.7 x 10(4) s-1.M-1) were determined from time-resolved activity assays. Tentoxin binding to the high affinity site was found independent on the catalytic state of the enzyme. The analysis of the kinetics of tentoxin binding on the low affinity site of the enzyme showed strong evidence for an interaction between this site and the nucleotide binding sites and revealed a complex relationship between the catalytic state and the reactivation process. New catalytic states of CF1 devoid of epsilon-subunit were detected: a transient overstimulated state, and a dead end complex unable to bind a second tentoxin molecule. Our experiments led to a kinetic model for the reactivation phenomenon for which rate constants were determined. The implications of this model are discussed in relation to the previous mechanistic hypotheses on the effect of tentoxin. PMID- 9873025 TI - A discrete three-amino acid segment (LVI) at the C-terminal end of kinase impaired ErbB3 is required for transactivation of ErbB2. AB - ErbB3 is unique among other members of the receptor tyrosine kinase family of growth factor receptors in that its kinase domain is enzymatically impaired. This renders it incapable of transducing a signal in response to ligand binding. However, in conjunction with ErbB2, ErbB3 is a potent mediator of signaling by the growth factor heregulin. Heregulin binding to ErbB3 induces formation of a heterodimeric complex with ErbB2, and this results in transactivation of the ErbB2 kinase. Although interaction between the extracellular domains of these receptors is an essential part of this process, it was not clear whether interaction between the cytoplasmic domains is also necessary for transactivation. By examining the abilities of a series of cytoplasmic domain mutants of ErbB3 to activate ErbB2, we have found a discrete sequence of three amino acid residues (LVI), located at the carboxyl-terminal end of the impaired ErbB3 kinase region, that is obligatory for transactivation. We conclude that formation of a functional ErbB2-ErbB3 signaling complex requires the presence of a specific structural feature within the ErbB3 cytoplasmic domain and suggest that ErbB2 transactivation results from a physical interaction between the cytoplasmic domains of these receptors. PMID- 9873026 TI - Identification and characterization of the gonadotropin-releasing hormone response elements in the mouse gonadotropin-releasing hormone receptor gene. AB - The response of the pituitary gonadotrope to gonadotropin-releasing hormone (GnRH) correlates directly with the concentration of GnRH receptors (GnRHR) on the cell surface, which is mediated in part at the level of GnRHR gene expression. Several hormones have been implicated in this regulation, most notably GnRH itself. Despite these observations and the central role that GnRH is known to play in reproductive development and function, the molecular mechanism(s) by which GnRH regulates transcription of the GnRHR gene has not been well elucidated. Previous studies in this laboratory have identified and partially characterized the promoter region of the mouse GnRHR gene and demonstrated that the regulatory elements for tissue-specific expression as well as for GnRH regulation are present within the 1.2-kilobase 5'-flanking sequence. By using deletion and mutational analysis as well as functional transfection studies in the murine gonadotrope-derived alphaT3-1 cell line, we have localized GnRH responsiveness of the mouse GnRHR gene to two DNA sequences at -276/-269 (designated Sequence Underlying Responsiveness to GnRH-2 (SURG-2), which contains the consensus sequence for the activating protein-1-binding site) and -292/-285 (a novel element designated SURG-1), and demonstrated that this response is mediated via protein kinase C. By using the electrophoretic mobility shift assay, we further demonstrate that a member(s) of the Fos/Jun heterodimer superfamily is responsible in part for the DNA-protein complexes formed on SURG-2, using alphaT3 1 nuclear extracts. These data define a bipartite GnRH response element in the mouse GnRHR 5'-flanking sequence and suggest that the activating protein-1 complex plays a central role in conferring GnRH responsiveness to the murine GnRHR gene. PMID- 9873027 TI - Extremely thermostable serine-type protease from Aquifex pyrophilus. Molecular cloning, expression, and characterization. AB - A gene encoding a serine-type protease has been cloned from Aquifex pyrophilus using a sequence tag containing the consensus sequence of proteases as a probe. Sequence analysis of the cloned gene reveals an open reading frame of 619 residues that has three canonical residues (Asp-140, His-184, and Ser-502) that form the catalytic site of serine-type proteases. The size of the mature form (43 kDa) and its localization in the cell wall fraction indicate that both the NH2- and COOH-terminal sequences of the protein are processed during maturation. When the cloned gene is expressed in Escherichia coli, it is weakly expressed as active and processed forms. The pH optimum of this protease is very broad, and its activity is completely inactivated by phenylmethylsulfonyl fluoride. The half life of the protein is 6 h at 105 degreesC, suggesting that it is one of the most heat-stable proteases. The cysteine residues in the mature form may form disulfide bonds that are responsible for the strong stability of this protease, because the thermostability of the protein is significantly reduced in the presence of reducing reagent. PMID- 9873028 TI - Regulation of gamma-aminobutyric acid (GABA) transporters by extracellular GABA. AB - gamma-Aminobutyric acid (GABA) transporters on neurons and glia at or near the synapse function to remove GABA from the synaptic cleft. Recent evidence suggests that GABA transporter function can be regulated, although the initial triggers for such regulation are not known. One hypothesis is that transporter function is modulated by extracellular GABA concentration, thus providing a feedback mechanism for the control of neurotransmitter levels at the synapse. To test this hypothesis, GABA uptake assays were performed on primary dissociated rat hippocampal cultures that endogenously express GABA transporters and on mammalian cells stably expressing the cloned rat brain GABA transporter GAT1. In both experimental systems, extracellular GABA induces chronic changes in GABA transport that occur in a dose-dependent and time-dependent manner. In addition to GABA, ACHC and nipecotic acid, both substrates of GAT1, up-regulate transport; GAT1 transport inhibitors that are not transporter substrates down-regulate transport. These changes occur in the presence of blockers of both GABAA and GABAB receptors, occur in the presence of protein synthesis inhibitors, and are not influenced by intracellular GABA. Surface biotinylation experiments reveal that the increase in transport is correlated with an increase in surface transporter expression. This increase in surface expression is due, at least in part, to a slowing of GAT1 internalization in the presence of extracellular GABA. These data suggest that the GABA transporter fine-tunes its function in response to extracellular GABA and would act to maintain a constant level of neurotransmitter at the synaptic cleft. PMID- 9873029 TI - Mutational analysis demonstrates that ClC-4 and ClC-5 directly mediate plasma membrane currents. AB - ClC-4 and ClC-5, together with ClC-3, form a distinct branch of the CLC chloride channel family. Although ClC-5 was shown to be mainly expressed in endocytotic vesicles, expression of ClC-5 in Xenopus oocytes elicited chloride currents. We now show that ClC-4 also gives rise to strongly outwardly rectifying anion currents when expressed in oocytes. They closely resemble ClC-5 currents with which they share a NO3- > Cl- > Br- > I- conductance sequence that differs from that reported for the highly homologous ClC-3. Both ClC-4 and ClC-5 currents are reduced by lowering extracellular pH. We could measure similar currents after expressing either channel in HEK293 cells. To demonstrate that these currents are directly mediated by the channel proteins, we introduced several point mutations that change channel characteristics. In ClC-5, several point mutations alter the kinetics of activation but leave macroscopic rectification and ion selectivity unchanged. A mutation (N565K) equivalent to a mutation reported to have profound effects on ClC-3 does not have similar effects on ClC-5. Moreover, a mutation at the end of D2 (S168T in ClC-5) changes ion selectivity, and a mutation at the end of D3 (E211A in ClC-5 and E224A in ClC-4) changes voltage dependence and ion selectivity. This shows that ClC-4 and ClC-5 can directly mediate plasma membrane currents. PMID- 9873030 TI - Demonstration of a direct interaction between residue 22 in the carboxyl-terminal half of secretin and the amino-terminal tail of the secretin receptor using photoaffinity labeling. AB - An understanding of the molecular basis of hormonal activation of receptors provides important insights for drug design. Toward this end, intrinsic photoaffinity labeling is a powerful tool to directly identify the ligand-binding domain. We have developed a new radioiodinatable agonist ligand of the secretin receptor that incorporates a photolabile p-benzoyl-L-phenylalanine (Bpa) into the position of Leu22 and have utilized this to identify the adjacent receptor domain. The rat [Tyr10,Bpa22]secretin-27 probe was a fully efficacious agonist, with a potency to stimulate cAMP accumulation by Chinese hamster ovary SecR cells similar to that of natural secretin (EC50 = 68 +/- 22 pM analogue and 95 +/- 25 pM secretin). It bound specifically and with high affinity (Ki = 5.0 +/- 1.1 nM) and covalently labeled the Mr = 57,000-62,000 secretin receptor. Cyanogen bromide cleavage of the receptor yielded a major labeled fragment of apparent Mr = 19,000 that shifted to Mr = 9,000 after deglycosylation. This was most consistent with either of two glycosylated domains within the amino-terminal tail of the receptor. Immunoprecipitation with antibody directed to epitope tags incorporated into each of the candidate domains established that the fragment at the amino terminus of the receptor was the site of labeling. This was further localized to the amino-terminal 30 residues of the receptor by additional proteolysis of this fragment with endoproteinase Lys-C. This provides the first direct demonstration of a contact between a secretin-like agonist and its receptor and will contribute a useful constraint to the modeling of this interaction. PMID- 9873031 TI - A new topological model of the cardiac sarcolemmal Na+-Ca2+ exchanger. AB - The current topological model of the Na+-Ca2+ exchanger consists of 11 transmembrane segments with extracellular loops a, c, e, g, i, and k and cytoplasmic loops b, d, f, h, and j. Cytoplasmic loop f, which plays a role in regulating the exchanger, is large and separates the first five from the last six transmembrane segments. We have tested this topological model by mutating residues near putative transmembrane segments to cysteine and then examining the effects of intracellular and extracellular applications of sulfhydryl-modifying reagents on exchanger activity. To aid in our topological studies, we also constructed a cysteineless Na+-Ca2+ exchanger. This mutant is fully functional in Na+ gradient-dependent 45Ca2+ uptake measurements and displays wild-type regulatory properties. It is concluded that the 15 endogenous cysteine residues are not essential for either activity or regulation of the exchanger. Our data support the current model by placing loops c and e at the extracellular surface and loops d, j, and l at the intracellular surface. However, the data also support placing Ser-788 of loop h at the extracellular surface and Gly-837 of loop i at the intracellular surface. To account for these data, we propose a revision of the model that places transmembrane segment 6 in cytoplasmic loop f. Additionally, we propose that putative transmembrane segment 9 does not span the membrane, but may form a "P-loop"-like structure. PMID- 9873032 TI - Differences in the regulation of fibroblast contraction of floating versus stressed collagen matrices. AB - To learn more about the regulation of contraction of collagen matrices by fibroblasts, we compared the ability of lysophosphatidic acid (LPA) and platelet derived growth factor (PDGF) to stimulate contraction of floating and stressed collagen matrices. In floating collagen matrices, PDGF and LPA stimulated contraction with similar kinetics, but appeared to utilize complementary signaling pathways since contraction obtained by the combination of growth factors exceeded that observed with saturating concentrations of either alone. The PDGF-simulated pathway was selectively inhibited by the protein kinase inhibitor KT5926. In stressed collagen matrices, PDGF and LPA stimulated contraction with different kinetics, with LPA acting rapidly and PDGF acting only after an approximately 1-h lag period. Pertussis toxin, known to block signaling through the Gi class of heterotrimeric G-proteins, inhibited LPA-stimulated contraction of floating but not stressed matrices, suggesting that LPA-stimulated contraction depends on receptors coupled to different G-proteins in floating and stressed matrices. On the other hand, the Rho inhibitor C3 exotransferase blocked contraction of both floating and stressed collagen matrices. These results suggest the possibility that distinct signaling mechanisms regulate contraction of floating and stressed collagen matrices. PMID- 9873033 TI - Purification, cloning, and characterization of the 16 S RNA m2G1207 methyltransferase from Escherichia coli. AB - The methyltransferase that forms m2G1207 in Escherichia coli small subunit rRNA has been purified, cloned, and characterized. The gene was identified from the N terminal sequence of the purified enzyme as the open reading frame yjjT (SWISS PROT accession number ). The gene, here renamed rsmC in view of its newly established function, codes for a 343-amino acid protein that has homologs in prokaryotes, Archaea, and possibly also in lower eukaryotes. The enzyme reacted well with 30 S subunits reconstituted from 16 S RNA transcripts and 30 S proteins but was almost inactive with the corresponding free RNA. By hybridization and protection of appropriate segments of 16 S RNA that had been extracted from 30 S subunits methylated by the enzyme, it was shown that of the three naturally occurring m2G residues, only m2G1207 was formed. Whereas close to unit stoichiometry of methylation could be achieved at 0.9 mM Mg2+, both 2 mM EDTA and 6 mM Mg2+ markedly reduced the level of methylation, suggesting that the optimal substrate may be a ribonucleoprotein particle less structured than a 30 S ribosome but more so than free RNA. PMID- 9873034 TI - Antifungal imidazoles block assembly of inducible NO synthase into an active dimer. AB - Cytokine-inducible nitric oxide synthase (iNOS) is a homodimeric enzyme that generates nitric oxide (NO) and L-citrulline from L-arginine (L-Arg) and O2. The N-terminal oxygenase domain (amino acids 1-498; iNOSox) in each subunit binds heme, L-Arg, and tetrahydrobiopterin (H4B), is the site of NO synthesis, and is responsible for the dimeric interaction, which must occur to synthesize NO. In both cells and purified systems, iNOS dimer assembly is promoted by H4B, L-Arg, and L-Arg analogs. We examined the ability of imidazole and N-substituted imidazoles to promote or inhibit dimerization of heme-containing iNOSox monomers, or to affect iNOS dimerization in cells. Imidazole, 1-phenylimidazole, clotrimazole, and miconazole all bound to the iNOSox monomer heme iron. Imidazole and 1-phenylimidazole promoted iNOSox dimerization, whereas clotrimazole (30 microM) and miconazole (15 microM) did not, and instead inhibited dimerization normally promoted by L-Arg and H4B. Clotrimazole also bound to iNOSox dimers in the absence of L-Arg and H4B and caused their dissociation. When added to cells expressing iNOS, clotrimazole (50 microM) had no effect on iNOS protein expression but almost completely inhibited its dimerization and consequent NO synthesis over an 8-h culture period, without affecting calmodulin interaction with iNOS. Thus, imidazoles can promote or inhibit dimerization of iNOS both in vitro and in cells, depending on their structure. Bulky imidazoles like clotrimazole block NO synthesis by inhibiting assembly of the iNOS dimer, revealing a new means to control cellular NO synthesis. PMID- 9873035 TI - Interleukin-1-induced nuclear factor-kappaB-IkappaBalpha autoregulatory feedback loop in hepatocytes. A role for protein kinase calpha in post-transcriptional regulation of ikappabalpha resynthesis. AB - The IkappaB inhibitors regulate the activity of the potent transcription factor nuclear factor-kappaB (NF-kappaB). Following signal-induced IkappaB proteolysis, NF-kappaB translocates into the nucleus to activate transcription of target genes, including IkappaBalpha itself, initiating the "NF-kappaB-IkappaBalpha autoregulatory feedback loop." Upon IkappaBalpha resynthesis, NF-kappaB is subsequently inactivated and redistributed back into the cytoplasm. We have previously reported a robust NF-kappaB-IkappaBalpha autoregulatory feedback loop in HepG2 hepatocytes. Sixty minutes after tumor necrosis factor (TNF-alpha) stimulation, IkappaBalpha is resynthesized to approximately 2-fold greater level than in control cells and completely inhibits NF-kappaB binding. Here we investigate the mechanism for IkappaBalpha resynthesis comparing the effect of stimulation of TNF-alpha with that of interleukin-1 (IL-1alpha). Although either TNF-alpha or IL-1alpha stimulation of protein kinase C (PKC)-down-regulated cells equivalently induces NF-kappaB translocation, the kinetics of IkappaBalpha resynthesis is slowed. Moreover, pretreatment with selective calcium-dependent PKC inhibitors selectively slowed the kinetics of the IL-1alpha-induced overshoot without affecting that produced by TNF-alpha. Down-regulation of PKCalpha by antisense phosphorothioate oligonucleotides and expression vectors selectively blocked the IL-1alpha-induced IkappaBalpha overshoot. In the absence of PKCalpha, although IL-1alpha induced similar amounts of IkappaBalpha transcription and changes in steady-state mRNA, a greater component of IkappaBalpha mRNA was retained in the nucleus. These data indicate a selective role for PKCalpha in IL 1alpha-induced IkappaBalpha resynthesis, which is mediated, at least in part, by post-transcriptional control of mRNA export. PMID- 9873036 TI - The influence of chromosomal location on the expression of two transgenes in mice. AB - We have generated mice having a single copy of the human haptoglobin gene (Hp2), driven by its natural promoter, and a neomycin resistance gene (Neo), driven by a herpes simplex thymidine kinase promoter with polyoma enhancers, inserted into two defined chromosomal locations, the Hprt locus on the X-chromosome and the apolipoprotein (apo) AI-CIII gene cluster on chromosome 9. The haptoglobin promoter is highly specialized in its tissue of action; the viral promoter has few restrictions. The apoAI-CIII gene is naturally active in only two tissues, whereas the Hprt gene region is ubiquitously active. Expression of both transgenes at substantial levels was achieved only (a) when the transgenes were inserted into the genome close to a known tissue-specific enhancer/locus control region in the apoAI-CIII gene cluster, and (b) when known conditions for function of their promoters were met. The specificities of the two chromosomal regions and of the two promoters are preserved, but their interactions are not specific. We conclude that transgenes are affected by locus-related enhancers in the same manner as nearby endogenous genes. Our experiments reinforce the usefulness of using gene targeting to direct single-copy transgenes to appropriate chromosomal locations. PMID- 9873037 TI - Agonists and inverse agonists for the herpesvirus 8-encoded constitutively active seven-transmembrane oncogene product, ORF-74. AB - A number of CXC chemokines competed with similar, nanomolar affinity against 125I interleukin-8 (IL-8) binding to ORF-74, a constitutively active seven transmembrane receptor encoded by human herpesvirus 8. However, in competition against 125I-labeled growth-related oncogene (GRO)-alpha, the ORF-74 receptor was highly selective for GRO peptides, with IL-8 being 10,000-fold less potent. The constitutive stimulating activity of ORF-74 on phosphatidylinositol turnover was not influenced by, for example, IL-8 binding. In contrast, GRO peptides acted as potent agonists in stimulating ORF-74 signaling, whereas IP-10 and stromal cell derived factor-1alpha surprisingly acted as inverse agonists. These peptides had similar pharmacological properties with regard to enhancing or inhibiting, respectively, the stimulatory effect of ORF-74 on NIH-3T3 cell proliferation. Construction of a high affinity zinc switch through introduction of two His residues at the extracellular end of transmembrane segment V enabled Zn2+ to act as a prototype non-peptide inverse agonist, which eliminated the constitutive signaling. It is concluded that ORF-74, which is believed to be causally involved in the formation of highly vascularized tumors, has been optimized for agonist and inverse agonist modulation by the endogenous angiogenic GRO peptides and angiostatic IP-10 and stromal cell-derived factor-1alpha, respectively. ORF-74 could serve as a target for the development of non-peptide inverse agonist drugs as demonstrated by the effect of Zn2+ on the metal ion site-engineered receptor. PMID- 9873038 TI - Sequence-specific DNA cleavage by Fe2+-mediated fenton reactions has possible biological implications. AB - Preferential cleavage sites have been determined for Fe2+/H2O2-mediated oxidations of DNA. In 50 mM H2O2, preferential cleavages occurred at the nucleoside 5' to each of the dG moieties in the sequence RGGG, a sequence found in a majority of telomere repeats. Within a plasmid containing a (TTAGGG)81 human telomere insert, 7-fold more strand breakage occurred in the restriction fragment with the insert than in a similar-sized control fragment. This result implies that telomeric DNA could protect coding DNA from oxidative damage and might also link oxidative damage and iron load to telomere shortening and aging. In micromolar H2O2, preferential cleavage occurred at the thymidine within the sequence RTGR, a sequence frequently found to be required in promoters for normal responses of many procaryotic and eucaryotic genes to iron or oxygen stress. Computer modeling of the interaction of Fe2+ with RTGR in B-DNA suggests that due to steric hindrance with the thymine methyl, Fe2+ associates in a specific manner with the thymine flipped out from the base stack so as to allow an octahedrally oriented coordination of the Fe2+ with the three purine N7 residues. Fe2+ dependent changes in NMR spectra of duplex oligonucleotides containing ATGA versus those containing AUGA or A5mCGA were consistent with this model. PMID- 9873039 TI - The platelet high affinity binding site for thrombin mimics hirudin, modulates thrombin-induced platelet activation, and is distinct from the glycoprotein Ib-IX V complex. AB - The platelet high affinity binding site for thrombin appears to be described by a classical receptor-ligand interaction that is distinct from the platelet thrombin receptor/substrate, PAR-1. However, the identification and function of the high affinity binding site with respect to its physiological importance have continued to elude investigators. Prior studies using two mutant thrombins suggested that thrombin interaction with the platelet high affinity binding site is mediated through an extensive portion of the thrombin molecule involving residues within the substrate binding pocket and the anion binding exosite (Leong, L., Henriksen, R. A., Kermode, J. C., Rittenhouse, S. E., and Tracy, P. B. (1992) Biochemistry 31, 2567-2575) and may mimic a thrombin-hirudin interaction. To test this hypothesis, an anti-hirudin peptide antibody (anti-hirpeptide Ab) was raised against a peptide mimicking the COOH terminus of hirudin. The Ab recognized adherent platelets and those in suspension as determined by enzyme-linked immunosorbent assay and immunofluorescence microscopy, respectively. 125I Thrombin binding to platelets was inhibited in the presence of the anti hirpeptide Ab in a dose-dependent manner with maximal inhibition >90%. Analyses of data from binding studies of 125I-thrombin to platelets at a fixed Ab concentration indicated that the anti-hirpeptide Ab inhibited the high affinity binding interaction exclusively. In addition, thrombin-induced increases in platelet [Ca2+]i were enhanced by blocking the high affinity binding site with the Ab due to redistribution of the agonist to PAR-1. Thrombin Quick I-induced platelet calcium mobilization was unaffected by the presence of the Ab, consistent with the inability of thrombin Quick I to bind to the high affinity site. Even though glycoprotein (GP) Ib contains a hirudin-like region within the alpha subunit, the postulated high affinity binding site, direct binding of 125I thrombin could not be demonstrated to transfected Chinese hamster ovary and L cells expressing the GP Ib-IX-V complex. Furthermore, an anti-GP Ib Ab, raised to the peptide region proposed as the thrombin high affinity site, did not enhance thrombin-induced platelet calcium mobilization. The anti-hirpeptide Ab recognized a population of platelet membrane proteins distinct from PAR-1 and GP Ib by three color immunofluorescence using confocal microscopy. These combined studies demonstrate that the high affinity binding site for thrombin is a unique platelet protein distinct from GP Ib which modulates the effective thrombin concentration localized at the human platelet surface. PMID- 9873040 TI - Cutis laxa arising from frameshift mutations in exon 30 of the elastin gene (ELN). AB - Congenital cutis laxa, a rare syndrome with marked skin laxity and pulmonary and cardiovascular compromise, is due to defective elastic fiber formation. In several cases, skin fibroblast tropoelastin production is markedly reduced yet reversed in vitro by transforming growth factor-beta treatment. We previously showed that this reversal was due to elastin mRNA stabilization in one cell strain, and here this behavior was confirmed in skin fibroblasts from two generations of a second family. cDNA sequencing and heteroduplex analysis of elastin gene transcripts from three fibroblast strains in two kindreds now identify two frameshift mutations (2012DeltaG and 2039DeltaC) in elastin gene exon 30, thus leading to missense C termini. No other mutations were present in the ELN cDNA sequences of all three affected individuals. Transcripts from both alleles in each kindred were unstable and responsive to transforming growth factor-beta. Exons 22, 23, 26A, and 32 were always absent. Since exon 30 underwent alternative splicing in fibroblasts, we speculate that a differential splicing pattern could conceivably lead to phenotypic rescue. These two dominant acting, apparently de novo mutations in the elastin gene appear to be responsible for qualitative and quantitative defects in elastin, resulting in the cutis laxa phenotype. PMID- 9873041 TI - Regulation of fas-ligand expression during activation-induced cell death in T lymphocytes via nuclear factor kappaB. AB - T cell receptor engagement activates transcription factors important for cytokine gene regulation. Additionally, this signaling pathway also leads to activation induced apoptosis in T lymphocytes that is dependent on FasL transcription and expression. Here we demonstrate that nuclear factor kappaB (NF-kappaB), which is involved in the transcriptional regulation of many cytokine genes expressed in activated lymphocytes, also plays a role in T cell activation-induced FasL expression. Inhibition of NF-kappaB activity in a T cell hybridoma leads to decreased FasL expression and apoptosis upon T cell receptor stimulation. We identified the NF-kappaB site in the FasL promoter that contributes to such regulation. Co-expression of p65 (Rel A) with the FasL promoter enhanced its activity, and co-expression of IkappaB dramatically inhibited the inducible promoter activity. In contrast, the transcription factor AP-1 is not required for activation-induced FasL promoter activity. These results define a role for NF kappaB in mediating FasL expression during T cell activation. PMID- 9873042 TI - Kalirin inhibition of inducible nitric-oxide synthase. AB - Nitric oxide (NO) acts as a neurotransmitter. However, excess NO produced from neuronal NO synthase (nNOS) or inducible NOS (iNOS) during inflammation of the central nervous system can be neurotoxic, disrupting neurotransmitter and hormone production and killing neurons. A screen of a hippocampal cDNA library showed that a unique region of the iNOS protein interacts with Kalirin, previously identified as an interactor with a secretory granule peptide biosynthetic enzyme. Kalirin associates with iNOS in vitro and in vivo and inhibits iNOS activity by preventing the formation of iNOS homodimers. Expression of exogenous Kalirin in pituitary cells dramatically reduces iNOS inhibition of ACTH secretion. Thus Kalirin may play a neuroprotective role during inflammation of the central nervous system by inhibiting iNOS activity. PMID- 9873043 TI - Adenovirus-mediated knockout of a conditional glucokinase gene in isolated pancreatic islets reveals an essential role for proximal metabolic coupling events in glucose-stimulated insulin secretion. AB - The relationship between glucokinase (GK) and glucose-stimulated metabolism, and the potential for metabolic coupling between beta cells, was examined in isolated mouse islets by using a recombinant adenovirus that expresses Cre recombinase (AdenoCre) to inactivate a conditional GK gene allele (gklox). Analysis of AdenoCre-treated islets indicated that the gklox allele in approximately 30% of islet cells was converted to a nonexpressing variant (gkdel). This resulted in a heterogeneous population of beta cells where GK was absent in some cells. Quantitative two-photon excitation imaging of NAD(P)H autofluorescence was then used to measure glucose-stimulated metabolic responses of individual islet beta cells from gklox/lox mice. In AdenoCre-infected islets, approximately one-third of the beta cells showed markedly lower NAD(P)H responses. These cells also exhibited glucose dose responses consistent with the loss of GK. Glucose dose responses of the low-responding cells were not sigmoidal and reached a maximum at approximately 5 mM glucose. In contrast, the normal response cells showed a sigmoidal response with an KcatS0.5 of approximately 8 mM. These data provide direct evidence that GK is essential for glucose-stimulated metabolic responses in beta cells within intact islets and that intercellular coupling within the islet plays little or no role in glucose-stimulated metabolic responses. PMID- 9873044 TI - Ligand-independent activation of the glucocorticoid receptor by beta2-adrenergic receptor agonists in primary human lung fibroblasts and vascular smooth muscle cells. AB - The glucocorticoid receptor (GR) is a ubiquitously expressed transcription factor present in most cell types. Upon ligand binding, the GR is activated and translocates into the nucleus where it transmits the anti-inflammatory actions of glucocorticoids. Here, we describe the ligand-independent activation of GR by the beta2-adrenergic receptor (beta2-AR) agonists, salbutamol and salmeterol, in primary human lung fibroblasts and vascular smooth muscle cells. Immunohistochemistry demonstrated expression of GR and the beta2-AR by fibroblasts and vascular smooth muscle cells. Treatment of the cells with the beta2-AR agonists, salbutamol or salmeterol, resulted in translocation of GR into the nucleus beginning at 30 min, as shown by immunohistochemistry and Western blotting of cytosolic and nuclear cell extracts. In comparison, activation of GR induced by the corticosteroids dexamethasone and fluticasone occurred at the same time after treatment (30 min) but resulted in a more complete depletion of GR from the cytosolic compartment. Electrophoretic mobility shift assays confirmed that nuclear GR, activated by both beta2-AR agonists and glucocorticoids, actively bound to the GR consensus sequence (GR element). Functional activation of the GR was confirmed by a Luciferase reporter gene assay, using a GR driven promoter fragment from the p21((WAF1/CIP1)) gene. The effects of the beta2-AR agonists, salbutamol and salmeterol, were dependent upon binding to the beta2-AR, because blocking of beta2-AR with propranolol abrogated GR activation. GR activation appeared to involve cAMP. In summary, these data show that beta2-AR agonists are potent activators of GR. Ligand-independent activation of GR by beta2-AR agonists may substantially mediate the anti-inflammatory actions of these drugs observed in vitro and in vivo. PMID- 9873045 TI - Glucose stimulates translocation of the homeodomain transcription factor PDX1 from the cytoplasm to the nucleus in pancreatic beta-cells. AB - One of the mechanisms whereby glucose stimulates insulin gene transcription in pancreatic beta-cells involves activation of the homeodomain transcription factor PDX1 (pancreatic/duodenal homeobox-1) via a stress-activated pathway involving stress-activated protein kinase 2 (SAPK2, also termed RK/p38, CSBP, and Mxi2). In the present study we show, by Western blotting and electrophoretic mobility shift assay, that in human islets of Langerhans incubated in low glucose (3 mM) PDX1 exists as an inactive 31-kDa protein localized exclusively in the cytoplasm. Transfer of the islets to high (16 mM) glucose results in rapid (within 10 min) conversion of PDX1 to an active 46-kDa form that was present predominantly in the nucleus. Activation of PDX1 appears to involve phosphorylation, as shown by incorporation of 32Pi into the 46-kDa form of the protein. These effects of glucose could be mimicked by chemical stress (sodium arsenite), or by overexpression of SAPK2 in the beta-cell line MIN6. Overexpression of SAPK2 also stimulated PDX1-dependent transcription of a -50 to -250 region of the human insulin gene promoter linked to a firefly luciferase reporter gene. The effects of glucose were inhibited by the SAPK2 inhibitor SB 203580, and by wortmannin and LY 294002, which inhibit phosphatidylinositol 3-kinase, although the effects of stress (arsenite) were inhibited only by SB 203580. These results demonstrate that glucose regulates the insulin gene promoter through activation and nuclear translocation of PDX1 via the SAPK2 pathway. PMID- 9873046 TI - Neisseria gonorrhoeae that infect men have lipooligosaccharides with terminal N acetyllactosamine repeats. AB - Infectious Neisseria gonorrhoeae make relatively large lipooligosaccharides (LOS) that structurally resemble human glycosphingolipids. MS11mkC is an LOS variant of N. gonorrhoeae strain MS11 which was isolated from men at the onset of dysuria (Schneider, H., Griffiss, J. M., Boslego, J. W., Hitchcock, P. J., Zahos, K. M., and Apicella, M. A. (1991) J. Exp. Med. 174, 1601-1605). Delayed extraction matrix-assisted laser desorption and ionization and electrospray ionization mass spectrometry of O-deacylated MS11mkC LOS produced ions consistent with known LOS which have lacto-N-neotetraose (Galbeta1-->4GlcNAcbeta1-->3Galbeta1-->4Glc; paraglobosyl; monoclonal antibodies (mAbs) 1B2(+) and 06B4(+)) and GalNAc-->lacto N-neotetraose (gangliosyl; mAb 1-1-M+) oligosaccharides. Ion peaks for a larger LOS which also bound mAb 1B2 indicated the addition of a hexose (+162 Da) to gangliosyl LOS or the addition of a hexose and a N-acetylhexosamine (+365 Da) to paraglobosyl LOS. Analysis of HF-treated and O-deacylated LOS revealed three major components present in a phosphoethanolamine (PEA)0 and a PEA1 series. Digestion of MS11mkC LOS by beta-N-acetylhexosaminidase and beta-galactosidase, alone and sequentially, combined with mAb binding patterns, confirmed the presence of a nonreducing terminal repeating LacNAc ((Galbeta1-->4GlcNAc)2) on the largest LOS, rather than a parallel oligosaccharide structure. PMID- 9873047 TI - Cloning and characterization of RLPK, a novel RSK-related protein kinase. AB - A novel protein kinase whose activity can be stimulated by mitogen in vivo was cloned and characterized. The cDNA of this gene encodes an 802-amino acid protein (termed RLPK) with the highest homology (37% identity) to the two protein kinase families, p90(RSK) and p70(RSK). Like p90(RSR), but not p70(RSK), RLPK also contains two complete nonidentical protein kinase domains. RLPK mRNA is widely expressed in all human tissues examined and is enriched in the brain, heart, and placenta. In HeLa cells, transiently expressed epitope-tagged RLPK can be strongly induced by epidermal growth factor, serum, and phorbol 12-myristate 13 acetate, but only moderately up-regulated by tumor necrosis factor-alpha and other stress-related stimuli. The activity of RLPK stimulated by epidermal growth factor was not inhibited by several known protein kinase C inhibitors nor by rapamycin, a known specific inhibitor for p70(RSK), but could be inhibited by herbimycin A, a tyrosine kinase inhibitor, and partially inhibited by PD98059 or SB203580, inhibitors for the mitogen-activated protein kinase pathways. Recombinant RLPK possesses high phosphorylation activity toward histone 2B and the S6 peptide, RRRLSSLRA. Although purified recombinant RLPK can be phosphorylated by ERK2 and p38alpha in vitro, its activity is not affected by this phosphorylation. Moreover, the treatment of RLPK with acid phosphatase did not reduce its in vitro kinase activity. These data suggest that RLPK is structurally similar to previously isolated RSKs, but its regulatory mechanism may be distinct from either p70(RSK) or p90(RSK)s. PMID- 9873048 TI - Interleukin-3 induces the phosphorylation of a distinct fraction of bcl-2. AB - Bcl-2-related proteins (i.e. Bcl-2 and Bax) regulate the effector stage of apoptosis and can modulate the entry of quiescent cells into the cell cycle. Phosphorylation of Bcl-2 is presumed to modify its apoptosis-inhibitory function. By utilizing an interleukin-3 (IL-3)-dependent hematopoietic cell line, we examined the structural requirements of Bcl-2 phosphorylation and the correlation of this post-translational modification with its function. In the presence of IL 3, constitutively expressed Bcl-2 was phosphorylated on serine residue(s), and phosphorylated Bcl-2 lost its capacity to heterodimerize with Bax. Whereas the majority of Bcl-2 resided in mitochondria, phosphorylation only affected a minor pool of total Bcl-2 that selectively partitioned into a soluble fraction. Cytosolic targeting of Bcl-2 greatly increased its ratio of phosphorylation. Bcl 2 phosphorylation was reduced during IL-3 deprivation, and its phosphorylation was also delayed after transient cytokine deprivation. This pattern of phosphorylation temporally correlated with the accelerated exit and delayed reentry of Bcl-2-expressing cells into the cell cycle upon transient IL-3 deprivation and subsequent cytokine restimulation. Thus, IL-3-induced phosphorylation of a distinct pool of Bcl-2 may contribute to the inactivation of its antiproliferative function. PMID- 9873049 TI - Two isoforms of protein disulfide isomerase alter the dimerization status of E2A proteins by a redox mechanism. AB - We have shown previously that E2A helix-loop-helix proteins spontaneously form an intermolecular disulfide cross-link that is required for stable homodimer binding to DNA (Benezra, R. (1994) Cell 79, 1057-1067). These homodimers are important for the development of B lymphocytes but are not present in other cell lineages. We have purified two proteins that are capable of regulating the formation of this disulfide bond and found them to be members of the protein disulfide isomerase (PDI) family. By regulating the formation of the disulfide cross-link, these proteins are capable of regulating the dimerization state of E proteins. PDI-mediated reduction appears to dissociate E protein homodimers and favors heterodimer formation with other basic helix-loop-helix proteins in both a purified protein system and in cellular extracts. These studies suggest that PDI may play an important role in the regulation of E2A transcription factor dimerization and the development of the B lymphocyte lineage. PMID- 9873050 TI - Monocyte adherence induced by lipopolysaccharide involves CD14, LFA-1, and cytohesin-1. Regulation by Rho and phosphatidylinositol 3-kinase. AB - Mechanisms regulating lipopolysaccharide (LPS)-induced adherence to intercellular adhesion molecule (ICAM)-1 were examined using THP-1 cells transfected with CD14 cDNA (THP-1wt). THP-1wt adherence to ICAM-1 was LPS dose-related, time-dependent, and inhibited by antibodies to either CD14 or leukocyte function associated antigen (LFA)-1, but was independent of any change in the number of surface expressed LFA-1 molecules. A potential role for phosphatidylinositol (PI) 3 kinase (PI 3-kinase) in LPS-induced adherence was examined using the PI 3-kinase inhibitors LY294002 and Wortmannin. Both inhibitors selectively attenuated LPS induced, but not phorbol 12-myristate 13-acetate-induced adherence. Inhibition by these agents was unrelated to any changes in either LPS binding to or LFA-1 expression by THP-1wt cells. LPS-induced adherence was also abrogated in U937 cells transfected with a dominant negative mutant of of PI 3-kinase. Toxin B from Clostridium difficile, an inhibitor of the Rho family of GTP-binding proteins, abrogated both PI-3 kinase activation and adherence induced by LPS. Cytohesin-1, a phosphatidylinositol 3,4,5-triphosphate-regulated adaptor molecule for LFA-1 activation, was found to be expressed in THP-1wt cells. In addition, treatment of THP-1wt with cytohesin-1 antisense attenuated LPS-induced adherence. These findings suggest a model in which LPS induces adherence through a process of "inside-out" signaling involving CD14, Rho, and PI 3-kinase. This converts low avidity LFA-1 into an active form capable of increased binding to ICAM-1. This change in LFA-1 appears to be cytohesin-1-dependent. PMID- 9873051 TI - Regulation of translational effectors by amino acid and mammalian target of rapamycin signaling pathways. Possible involvement of autophagy in cultured hepatoma cells. AB - Amino acid deprivation of Chinese hamster ovary cells overexpressing human insulin receptors results in deactivation of p70 S6 kinase (p70) and dephosphorylation of eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), which become unresponsive to insulin; readdition of amino acids restores these responses in a rapamycin-sensitive manner, suggesting that amino acids and mammalian target of rapamycin signal through common effectors. Contrarily, withdrawal of medium amino acids from the hepatoma cell line H4IIE does not abolish the ability of insulin to stimulate p70 and 4E-BP1. The addition of 3 methyladenine (3MA) to H4IIE cells deprived of amino acids inhibited the increment in protein degradation caused by amino acid withdrawal nearly completely at 10 mM and also strongly inhibited the ability of insulin to stimulate p70 and 4E-BP1 at 10 mM. Treatment of H4IIE cells with 3MA did not alter the ability of insulin to activate tyrosine phosphorylation, phosphoinositide 3-kinase, or mitogen-activated protein kinase. In conclusion, the ability of H4IIE cells to maintain the insulin responsiveness of the mammalian target of rapamycin-dependent signaling pathways impinging on p70 and 4E-BP1 without exogenous amino acids reflects the generation of amino acids endogenously through a 3MA-sensitive process, presumably autophagy, a major mechanism of facultative protein degradation in liver. PMID- 9873052 TI - G4 DNA binding by LR1 and its subunits, nucleolin and hnRNP D, A role for G-G pairing in immunoglobulin switch recombination. AB - The immunoglobulin heavy chain switch regions contain multiple runs of guanines on the top (nontemplate) DNA strand. Here we show that LR1, a B cell-specific, duplex DNA binding factor, binds tightly and specifically to synthetic oligonucleotides containing G-G base pairs (KD Gln mutants did not significantly change the Kd or N. None of the mutations compromised the ability of the module to protect all-trans retinol from degradation. CONCLUSIONS: Our data suggest that the function of the conserved arginines in IRBP is fundamentally different from that of other retinoid-binding proteins. These residues do not appear to play a role in defining the specificity of the ligand-binding domain. Rather, Arg1005 appears to play a role in defining the capacity of the domain. Our data suggest that the binding site consists of a single hydrophobic cavity promiscuous for fatty acids and all-trans retinol. PMID- 9873069 TI - Familial subepithelial corneal amyloidosis (gelatinous drop-like corneal dystrophy): exclusion of linkage to lactoferrin gene. AB - PURPOSE: Because corneal tissue with familial subepithelial corneal amyloidosis (FSCA; gelatinous drop-like dystrophy of the cornea) contains lactoferrin the possibility that the FSCA gene was the human lactoferrin (hLF) gene was investigated. Due to contradictory published information we also mapped the hLF gene. METHODS: We mapped the hLF gene using a genomic clone of the entire hLF gene as a probe by fluorescence in situ hybridization (FISH). Utilizing PCR primers that are specific to the hLF gene, we also mapped the hLF via radiation somatic cell hybrid analysis. Linkage of the FSCA gene to the hLF gene was evaluated by genetic linkage analysis using polymorphic markers within and in the vicinity of the hLF gene. RESULTS: The hLF gene mapped to the short arm of chromosome 3 at 3p21. Linkage analysis using polymorphic markers for hLF and haplotype analysis of the 3p21 loci indicates that the FSCA gene is not linked to the 3p21 locus. CONCLUSIONS: The gene for FSCA is not the hLF gene in these families. PMID- 9873070 TI - Spatial and temporal expression of AP-1 responsive rod photoreceptor genes and bZIP transcription factors during development of the rat retina. AB - PURPOSE: The promoter region of the rod-specific beta subunit of cGMP PDE (beta PDE) and opsin genes contains highly conserved cis-acting elements, which include an AP-1 and/or Nrl response element (NRE: An extended AP-1 like sequence). Transactivation of AP-1 or NRE appears necessary to drive expression of these rod specific genes during adulthood, however, their role during development is relatively unknown. Therefore, we determined the spatial and temporal relationships between rod morphological and functional development, rod-specific gene expression, and expression of the bZIP transcription factors c-fos, junD and Nrl. METHODS: Retinas from 0-45 day old (PN0-45) dark- and light-adapted Long Evans rats were used. Morphological development was monitored by light and electron microscopy. Whole retinal trypsin-activated cGMP-PDE activity and rhodopsin content were measured biochemically. The expression of opsin, beta-PDE, c-fos, junD and Nrl mRNAs were determined by Northern blot analysis. The cellular localization of Nrl was examined with in situ hybridization. RESULTS: The mRNAs for opsin, beta-PDE and c-fos were observed at PN0-2, while cGMP-PDE activity and rhodopsin were detected first at PN5: coincident with rod outer segment development. The developmental pattern of cGMP-PDE activity and rhodopsin accumulation paralleled the expression of beta-PDE and opsin mRNA and all reached their maximal levels by PN45. Nrl expression, for all three transcripts found in the rat retina, was low on PN2 and reached its maximal level at PN14. The c-fos and Nrl expression preceded beta-PDE and opsin mRNA expression by 1-2 days. Nrl expression was detected first in the distal post-mitotic retina at PN5 and then in all nuclear layers during retinal development. Maximal expression shifted from the ganglion cells to the outer nuclear layer as the neural retina matured. In contrast, junD expression was highest at PN0 and declined to a stable level by PN10. CONCLUSIONS: Colocalization of Nrl and c-Fos suggests that expression of rod-specific genes, which utilize AP-1 or NRE sites in their promoter, could be regulated through the formation of Nrl-Fos dimers. We hypothesize that Nrl and c Fos play a fundamental role in the initiation and regulation of the rod-specific gene expression in developing and adult rod photoreceptors. PMID- 9873071 TI - Structure-function relationships in the four repeats of human interphotoreceptor retinoid-binding protein (IRBP). AB - PURPOSE: Interphotoreceptor retinoid-binding protein (IRBP) binds hydrophobic ligands in the interphotoreceptor space. Human IRBP consists of 1230 amino acids in four 300 amino acid long repeats. We asked: 1. Whether each of the four repeats can bind retinoids or fatty acids, 2. Whether each repeat can prevent retinol degradation in aqueous solutions, 3. Whether a ligand can stabilize the protein from thermal denaturation, 4. Whether the four repeats can be further classified into two groups. Our rationale was to make each repeat from the human cDNA and then examine structural and functional characteristics. METHODS: Individual repeats were produced in E. coli and the whole protein was expressed in baculovirus. Binding properties with all-trans-retinol were characterized by ligand fluorescence enhancement. The quenching of protein fluorescence by retinol, 9-cis-retinal, all-trans-retinoic acid, beta-ionine, alpha-ionine, trans parinaric acid, and DHA was also examined. Binding curves were analyzed by nonlinear regression. Prevention of retinol decomposition was measured by absorption spectroscopy. Circular dichroism was examined in the far UV range to study protein secondary structure and the near UV range to study ligand binding effects on the tryptophan environment. RESULTS: Temperature dependent denaturation suggests that EcR1 is the most stable of the four repeats. Each repeat possesses the capability of binding 9-cis-retinal, all-trans-retinol, all trans retinoic acid, docosahexaenoic acid, alpha- and beta-ionine, and trans parinaric acid. Protein fluorescence quenching by retinol and retinol fluorescence enhancement assays yielded similar binding parameters for each repeat. Each expressed repeat prevents the degradation of retinol in aqueous solutions. CONCLUSIONS: The data contrast with the idea that two or more repeats are needed to bind one molecule of ligand. Each repeat binds both retinoids and analogs, suggesting that each has multiple ligand binding sites or one binding site with affinity for different ligands. Together, the results suggest that each repeat retains all functions of the whole protein. However, there are distinguishing characteristics among the repeats in their ligand binding properties, though the four repeats cannot be classified into just two distinctive groups. Last, these data fit well with the current model of multiple binding sites in IRBP derived from quadruplication of an ancestral monomeric binding protein. PMID- 9873072 TI - Evolution of ITS1 rDNA in the Digenea (Platyhelminthes: trematoda): 3' end sequence conservation and its phylogenetic utility. AB - A comparison of ribosomal internal transcribed spacer 1 (ITS1) elements of digenetic trematodes (Platyhelminthes) including unidentified digeneans isolated from Cyathura carinata (Crustacea: Isopoda) revealed DNA sequence similarities at more than half of the spacer at its 3' end. Primary sequence similarity was shown to be associated with secondary structure conservation, which suggested that similarity is due to identity by descent and not chance. Using an analysis of apomorphies, the sequence data were shown to produce a distinct phylogenetic signal. This was confirmed by the consistency of results of different tree reconstruction methods such as distance approaches, maximum parsimony, and maximum likelihood. Morphological evidence additionally supported the phylogenetic tree based on ITS1 data and the inferred phylogenetic position of the unidentified digeneans of C. carinata met the expectations from known trematode life-cycle patterns. Although ribosomal ITS1 elements are generally believed to be too variable for phylogenetic analysis above the species or genus level, the overall consistency of the results of this study strongly suggests that this is not the case in digenetic trematodes. Here, 3' end ITS1 sequence data seem to provide a valuable tool for elucidating phylogenetic relationships of a broad range of phylogenetically distinct taxa. PMID- 9873074 TI - Getting in or out: early segregation between importers and exporters in the evolution of ATP-binding cassette (ABC) transporters. AB - ATP-binding cassette (ABC) systems, also called traffic ATPases, are found in eukaryotes and prokaryotes and almost all participate in the transport of a wide variety of molecules. ABC systems are characterized by a highly conserved ATPase module called here the ABC module, involved in coupling transport to ATP hydrolysis. We have used the sequence of one of the first representatives of bacterial ABC transporters, the MalK protein, to collect 250 closely related sequences from a nonredundant protein sequence database. The sequences collected by this objective method are all known or putative ABC transporters. After having eliminated short protein sequences and duplicates, the 197 remaining sequences were subjected to a phylogenetic analysis based on a mutational similarity matrix. An unrooted tree for these modules was found to display two major branches, one grouping all collected uptake systems and the other all collected export systems. This remarkable disposition strongly suggests that the divergence between these two functionally different types of ABC systems occurred once in the history of these systems and probably before the differentiation of prokaryotes and eukaryotes. We discuss the implications of this finding and we propose a model accounting for the generation and the diversification of ABC systems. PMID- 9873073 TI - Short inverted-repeat transposable elements in teleost fish and implications for a mechanism of their amplification. AB - Angel is the first miniature inverted-repeat transposable element (MITE) isolated from fish. Angel elements are imperfect palindromes with the potential to form stem-loop structures in vitro. Despite sequence divergence of elements of up to 55% within and between species, their inverted repeat structures have been maintained, implying functional importance. We estimate that there are about 10(3)-10(4) Angels scattered throughout the zebrafish genome, evidence that this family of transposable elements has been significantly amplified over the course of evolution. Angel elements and Xenopus MITEs carry common sequence motifs at their termini, indicating common origin and/or related mechanisms of transposition. We present a model in which MITEs take advantage of the basic cellular mechanism of DNA replication for their amplification, which is dependent on the characteristic inverted repeat structures of these elements. We propose that MITEs are genomic parasites that transpose via a DNA intermediate, which forms by a folding-back of a single strand of DNA, that borrow all of the necessary factors for their amplification from products encoded in the genomes in which they reside. DNA polymorphisms in different lines of zebrafish were detected by PCR using Angel-specific primers, indicating that such elements, combined with other transposons in vertebrate genomes, will be useful molecular tools for genome mapping and genetic analyses of mutations. PMID- 9873075 TI - Phylogeny of Japanese papilionid butterflies inferred from nucleotide sequences of the mitochondrial ND5 gene. AB - Phylogenetic relationships among the Japanese papilionid butterflies were analyzed by comparing 783 nucleotide sequences of the mitochondrial gene encoding NADH dehydrogenase subunit 5 (ND5). Phylogenetic trees of the representative species from each family in the superfamily Papilionoidea revealed that the species of the family Papilionidae and those of all other families formed distinct clusters, with a few species of the family Hesperiidae (Hesperioidea) as an outgroup. In the phylogenetic trees of most Japanese species of the family Papilionidae with Nymphalis xanthomelas (Nymphalidae) as an outgroup, the tribe Parnassiini (Parnassiinae) formed a cluster, and the rest formed the other cluster in which the tribe Zerynthiini (Parnassiinae) and the subfamily Papilioninae formed different subclusters. In the Papilioninae cluster, the tribes Troidini and Graphiini formed a subcluster, and the tribe Papilionini formed the other subcluster. These results generally agree with the traditional classification of the papilionid butterflies based on their morphological characteristics and support the proposed evolutionary genealogy of the butterflies based on their morphology, behavior, and larval host plants, except that the tribes Parnasiini and Zerynthiini (both Parnassiinae) are not in the same cluster. PMID- 9873077 TI - A phylogenetic assessment of the eukaryotic light-harvesting antenna proteins, with implications for plastid evolution. AB - The light-harvesting complexes (LHCs) are a superfamily of chlorophyll-binding proteins present in all photosynthetic eukaryotes. The Lhc genes are nuclear encoded, yet the pigment-protein complexes are localized to the thylakoid membrane and provide a marker to follow the evolutionary paths of plastids with different pigmentation. The LHCs are divided into the chlorophyll a/b-binding proteins of the green algae, euglenoids, and higher plants and the chlorophyll a/c-binding proteins of various algal taxa. This work examines the phylogenetic position of the LHCs from three additional taxa: the rhodophytes, the cryptophytes, and the chlorarachniophytes. Phylogenetic analysis of the LHC sequences provides strong statistical support for the clustering of the rhodophyte and cryptomonad LHC sequences within the chlorophyll a/c-binding protein lineage, which includes the fucoxanthin-chlorophyll proteins (FCP) of the heterokonts and the intrinsic peridinin-chlorophyll proteins (iPCP) of the dinoflagellates. These associations suggest that plastids from the heterokonts, haptophytes, cryptomonads, and the dinoflagellate, Amphidinium, evolved from a red algal-like ancestor. The Chlorarachnion LHC is part of the chlorophyll a/b binding protein assemblage, consistent with pigmentation, providing further evidence that its plastid evolved from a green algal secondary endosymbiosis. The Chlorarachnion LHC sequences cluster with the green algal LHCs that are predominantly associated with photosystem II (LHCII). This suggests that the green algal endosymbiont that evolved into the Chlorarachnion plastid was acquired following the emergence of distinct LHCI and LHCII complexes. PMID- 9873076 TI - Concordant evolution of a symbiont with its host insect species: molecular phylogeny of genus Glossina and its bacteriome-associated endosymbiont, Wigglesworthia glossinidia. AB - Many arthropods with restricted diets rely on symbiotic associations for full nutrition and fecundity. Tsetse flies (Diptera: Glossinidae) harbor three symbiotic organisms in addition to the parasitic African trypanosomes they transmit. Two of these microorganisms reside in different gut cells, while the third organism is harbored in reproductive tissues and belongs to the genus Wolbachia. The primary symbiont (genus Wigglesworthia glossinidia) lives in differentiated epithelial cells (bacteriocytes) which form an organ (bacteriome) in the anterior gut, while the secondary (S) symbionts are present in midgut cells. Here we have characterized the phylogeny of Wigglesworthia based on their 16S rDNA sequence analysis from eight species representing the three subgenera of Glossina: Austenina (=fusca group), Nemorhina (=palpalis group), and Glossina (=morsitans group). Independently, the ribosomal DNA internal transcribed spacer 2 (ITS-2) regions from these species were analyzed. The analysis of Wigglesworthia indicated that they form a distinct lineage in the gamma subdivision of Proteobacteria and display concordance with their host insect species. The trees generated by parsimony confirmed the monophyletic taxonomic placement of Glossina, where fusca group species formed the deepest branch followed by morsitans and palpalis groups, respectively. The placement of the species Glossina austeni by both the traditional morphological and biochemical criteria has been controversial. Results presented here, based on both the ITS-2 and the symbiont 16S rDNA sequence analysis, suggest that Glossina austeni should be placed into a separate fourth subgenus, Machadomyia, which forms a sister group relationship with the morsitans group species. PMID- 9873078 TI - Variable and invariable DNA repeat characters revealed by taxonprint approach are useful for molecular systematics. AB - A specially optimized restriction analysis of highly repetitive DNA elements, called DNA taxonprint, was applied for phylogenetic study of primates and lizards. It was shown that electrophoretic bands of DNA repeats revealed by the taxonprint technique have valuable properties for molecular systematics. Approximately half of taxonprint bands (TB) are invariable and do not disappear from the genomes during evolution or change spontaneously. Presumably these invariable bands are restriction fragments of dispersed DNA repeats. Another group represents variable taxonprint bands that differ even between closely related species. These variable bands are probably represented by tandem DNA repeats and could be used as species-specific markers. It was shown that taxonprint bands are independent characters since the appearance of a new taxonprint band does not change the previous band pattern. Phylogenetic reconstruction carried out on taxonprint data demonstrated that this approach could be of general utility for molecular systematics and species identification. PMID- 9873079 TI - Genetic characterization of plasmids containing genes encoding enzymes of leucine biosynthesis in endosymbionts (Buchnera) of aphids. AB - The prokaryotic endosymbionts (Buchnera) of aphids are known to provision their hosts with amino acids that are limiting in the aphid diet. Buchnera from the aphids Schizaphis graminum and Diuraphis noxia have plasmids containing leuABCD, genes that encode enzymes of the leucine biosynthetic pathway, as well as genes encoding proteins probably involved in plasmid replication (repA1 and repA2) and an open reading frame (ORF1) of unknown function. The newly reported plasmids closely resemble a plasmid previously described in Buchnera of the aphid Rhopalosiphum padi [Bracho AM, Martinez-Torres D, Moya A, Latorre A (1995) J Mol Evol 41:67-73]. Nucleotide sequence comparisons indicate conserved regions which may correspond to an origin of replication and two promoters, as well as inverted repeats, one of which resembles a rho-independent terminator. Phylogenetic analyses based on amino acid sequences of leu gene products and ORF1 resulted in trees identical to those obtained from endosymbiont chromosomal genes and the plasmid-borne trpEG. These results are consistent with a single evolutionary origin of the leuABCD-containing plasmid in a common ancestor of Aphididae and the lack of plasmid exchange between endosymbionts of different aphid species. Trees for ORF1 and repA (based on both nucleotides and amino acids) are used to examine the basis for leu plasmid differences between Buchnera of Thelaxes suberi and Aphididae. The most plausible explanation is that a single transfer of the leu genes to an ancestral replicon was followed by rearrangements. The related replicon in Buchnera of Pemphigidae, which lacks leuABCD, appears to represent the ancestral condition, implying that the plasmid location of the leu genes arose after the Pemphigidae diverged from other aphid families. This conclusion parallels previously published observations for the unrelated trpEG plasmid, which is present in Aphididae and absent in Pemphigidae. Recruitment of amino acid biosynthetic genes to plasmids has been ongoing in Buchnera lineages after the infection of aphid hosts. PMID- 9873080 TI - Variation in the pattern of nucleotide substitution across sites. AB - A model of nucleotide substitution that allows the transition/transversion rate bias to vary across sites was constructed. We examined the fit of this model using likelihood-ratio tests by analyzing 13 protein coding genes and 1 pseudogene. Likelihood-ratio testing indicated that a model that allows variation in the transition/transversion rate bias across sites provided a significant improvement in fit for most protein coding genes but not for the pseudogene. When the analysis was repeated with parameters estimated separately for first, second, and third codon positions, strong heterogeneity was uncovered for the first and second codon positions; the variation in the transition/transversion rate was generally weaker at the third codon position. The transition rate bias and branch lengths are underestimated when variation in the transition/transversion rate was not accommodated, suggesting that it may be important to accommodate variation in the pattern of nucleotide substitution for accurate estimation of evolutionary parameters. PMID- 9873083 TI - Ancient gene duplication and differential gene flow in plastid lineages: the GroEL/Cpn60 example. AB - Cryptomonads, small biflagellate algae, contain four different genomes. In addition to the nucleus, mitochondrion, and chloroplast is a fourth DNA containing organelle the nucleomorph. Nucleomorphs result from the successive reduction of the nucleus of an engulfed phototrophic eukaryotic endosymbiont by a secondary eukaryotic host cell. By sequencing the chloroplast genome and the nucleomorph chromosomes, we identified a groEL homologue in the genome of the chloroplast and a related cpn60 in one of the nucleomorph chromosomes. The nucleomorph-encoded Cpn60 and the chloroplast-encoded GroEL correspond in each case to one of the two divergent GroEL homologues in the cyanobacterium Synechocystis sp. PCC6803. The coexistence of divergent groEL/cpn60 genes in different genomes in one cell offers insights into gene transfer from evolving chloroplasts to cell nuclei and convergent gene evolution in chlorophyll a/b versus chlorophyll a/c/phycobilin eukaryotic lineages. PMID- 9873082 TI - Biased nucleotide composition of the genome of HERV-K related endogenous retroviruses and its evolutionary implications. AB - The human genome contains a large number of sequences that belong to the HERV-K family of human endogenous retroviruses. Most of these elements are likely remnants of ancient infections by ancestral exogenous retroviruses. To obtain further insight into the evolutionary history and molecular mechanisms responsible for the diversity of the human HERV-K elements, we analyzed several aspects of their genome structure. The nucleotide composition of the HERV-K genome was found to be highly biased and asymmetric, with an abundance of the A nucleotide in the viral (+) strand. A similar trend has been reported for the genomes of several exogenous retroviruses, with different nucleotides as the preferred building block. Other genome characteristics that were reported previously for actively replicating retroviruses are also apparent for the endogenous HERV-K virus. In particular, we observed suppression of the dinucleotide CpG, which represents potential methylation sites, and a strong preference for synonymous substitutions within the open reading frame of the reverse transcriptase (RT) enzyme. Furthermore, the mutational spectrum of the HERV-K RT enzyme was evaluated by nucleotide sequence comparison of 34 available elements. Interestingly, this analysis revealed a striking similarity with the mutational pattern of the HIV-1 RT enzyme, with a preference for G-to-A and C-to T transitions. It is proposed that the mutational bias of the HERV-K RT enzyme played a role in the shaping of this retroviral genome, which was actively replicating more than 30 million years ago. This effect can still be observed in the contemporary endogenous HERV-K elements. PMID- 9873081 TI - Intraspecies analysis: comparison of ITS sequence data and gene intron sequence data with breeding data for a worldwide collection of Gonium pectorale. AB - The morphologically uniform species Gonium pectorale is a colonial green flagellate of worldwide distribution. The affinities of 25 isolates from 18 sites on five continents were assessed by both DNA sequence comparisons and sexual compatibility. Complete sequences were obtained (i) for the internal transcribed spacer ITS-1 and ITS-2 regions of ribosomal DNA and (ii) for each of three single copy spliceosomal introns, two in a small G protein and one in the actin gene. ITS sequences appeared to homogenize sufficiently rapidly to behave as a single copy gene. Intron sequence differences between isolates in this species reached nucleotide substitution saturation, while ITS sequences did not. Parsimony and evolutionary distance analysis of the two types of DNA data gave essentially the same tree conformation. By all these criteria, the group of G. pectorale isolates fell into two main clades, A and B. Clade A, with isolates from four continents, was comprised of four subclades of quite closely related isolates, plus one strain of ambiguous affinity. Clade B was comprised of two subclades represented by South African and South American isolates, respectively; thus, only subclades of clade B showed geographical localization. With respect to mating, all isolates except one homothallic strain and one apparently sterile strain fell into either one or the other of two mating types. Pairings in all possible combinations revealed that isolates from the same site formed abundant zygotes, which germinated to produce new, sexually active organisms. Zygotes were also formed in many pairings of other combinations, including crosses of clade A with clade B organisms, but none of the latter produced viable germlings. The ability to mate and produce viable progeny that were themselves capable of sexual reproduction was restricted to members of subclades established on the basis of DNA sequence similarities. Thus, the grades of difference in both nuclear intron sequences and rDNA ITS sequences paralleled those observed in the sexual analysis. PMID- 9873084 TI - Phylogenetic studies of complete mitochondrial DNA molecules place cartilaginous fishes within the tree of bony fishes. AB - It is commonly acknowledged that cartilaginous fishes, Chondrichthyes, have a basal position among the Gnathostomata (jawed vertebrates). In order to explore this relationship we have sequenced the complete mitochondrial genome of the spiny dogfish, Squalus acanthias, and included it in a phylogenetic analysis together with a number of bony fishes and amniotes. The phylogenetic reconstructions placed the dogfish among the bony fishes. Thus, and contrary to the common view, the analyses have shown that the position of the sharks is not basal among the gnathostomes. The presently recognized phylogenetic position of the dogfish was identified irrespective of the outgroup used, echinoderms or agnathan fishes. The lungfish was the most basal gnathostome fish, while the teleosteans had an apical position in the piscine tree. A basal position of the dogfish among the gnathostomes was statistically rejected, but the phylogenetic relationship among the coelacanth, spiny dogfish, and teleosts was not conclusively resolved. The findings challenge the current theory that sharks and other chondrichthyans, if monophyletic, are the sister group to all other extant gnathostomes. The results open to question the status of several morphological characters commonly used in piscine phylogenetic reconstruction, most notably the presence versus absence of endochondral bone in the endoskeleton, the macromeric versus micromeric structure of the exoskeleton, and the presence/absence of swimbladder and/or lung. The study also confirmed recent findings demonstrating that the origin of the amniotes is deeper than the diversification of extant bony fishes. PMID- 9873085 TI - CD40L (CD154) expression in human liver allografts during chronic ductopenic rejection. AB - The CD40-CD40L (CD154) interaction plays a pivotal role in the effector mechanisms of allograft rejection. Blockade of the CD40/CD40L costimulatory pathway prevents the development of chronic allograft rejection in several animal transplant models. The relevance of in situ CD40 and CD40L expression in human liver allografts was assessed by immunohistochemistry during ductopenic chronic rejection (CR). In CR allograft specimens (n = 8), marked CD40L expression was detected on Kupffer cells (KCs) and sinusoidal macrophages with a unique centrilobular distribution (P <.001). The CD40L+ KCs and macrophages were shown to be CD68+ after immunohistochemical analysis of serial sections with anti-CD68 monoclonal antibody. Moderate staining of vascular and sinusoidal endothelial cells and mononuclear infiltrates was observed in some CR cases. These findings were in contrast to the absence of CD40L expression in controls (n = 11) consisting of stable liver allograft and normal liver tissue specimens. Only occasional CD40 expression in some cases of CR and controls was observed. In CR, CD40L (CD154) expression is manifested on KCs and macrophages. The present novel data show another important cellular source of CD40L expression and suggest a potential role of KCs/macrophages and CD40/CD40L costimulatory interactions in the pathogenesis of chronic rejection ductopenic liver allograft. PMID- 9873086 TI - In liver transplantation, T tube bile represents total bile flow: physiological and scintigraphic studies on biliary secretion of organic anions. AB - The present study was performed to clarify the recovery of hepatocellular uptake and the biliary secretion of bile acids during the first 14 days after orthotopic liver transplantation (OLT) and to determine the fraction of bile flow appearing outside through the T tube and entering the duodenum. Therefore, we determined primary and secondary bile acids in bile samples obtained from the T tube at day 5 after OLT, while the T tube was permanently open, and at days 10 and 14 after OLT, i.e., 4 and 9 days after closure of the T tube, respectively, thus restoring enterohepatic bile acid circulation. In addition, we performed hepatobiliary scintigraphy using technetium 99m-labeled [2,4,6 trimethyl-3-bromo]imino-diacetic acid (technetium 99m-BRIDA) in 12 patients between days 4 and 17 after OLT. Chromatographic analyses of biliary bile acids showed no secondary bile acids during the first 5 days after OLT, as opposed to 10 and 14 days after OLT when enterohepatic circulation was restored. Eleven patients with an uncomplicated postoperative course after OLT showed a significantly reduced hepatic uptake and biliary secretion of 99mTc-BRIDA during the first days after OLT with progressive recovery. One patient with an acute allograft rejection episode showed almost no uptake and only minimal secretion. The bile fraction appearing outside through the inserted T tube represented 94.6% +/- 6.2% of the injected 99mTc-BRIDA. We conclude that OLT results in markedly impaired hepatocellular uptake and biliary secretion of organic anions. Simultaneously, bile acid synthesis is significantly reduced, which, in addition, diminishes bile secretion of the graft. We show that T tube bile is a valid tool for bile physiological studies in patients in whom transplantation was successfully performed. PMID- 9873087 TI - Candida infection in pediatric liver transplant recipients. AB - A retrospective review of 100 liver transplantations in 98 children was performed to determine the incidence of infection caused by Candida organism in these patients and to identify risk factors that may predispose to serious fungal infection. Thirty-one infections caused by Candida organisms developed during the initial 28 days posttransplantation: 19 were definite invasive infections (one deep site or one positive blood culture), 2 were probable invasive infections (three superficial sites), and 10 were urinary tract infections. Eleven of 19 patients had fungemia or a disseminated infection (two noncontiguous deep organs involved and/or positive blood cultures) and 8 of 19 had peritoneal candidiasis. Infection caused by Candida organisms was a contributing factor to mortality in 7 of 21 patients (case fatality rate of 33%) with invasive infection. Risk factors that were predictive for invasive infection by univariate analysis included the following: pretransplantation antibiotic therapy, length of transplant operation, transfusion requirement, number of days in the intensive care unit, number of days intubated, number of concurrent bacterial infections, number of antibiotics administered, number of laparotomies performed posttransplantation, retransplantation, hepatic artery thrombosis, bile leaks, and renal and respiratory failure. By logistic regression analysis, bile leak, hepatic artery thrombosis, preoperative steroid use, transfusion requirement, and the number of days intubated were identified as independent risk factors for invasive infection caused by Candida organisms. The use of prophylactic antifungal agents in high risk patients may be important in reducing the serious morbidity and mortality associated with sepsis caused by Candida organisms in pediatric liver transplant recipients. PMID- 9873088 TI - Rimantadine for treatment of hepatitis C infection in liver transplant recipients. AB - Hepatitis C recurrence after liver transplantation is a serious problem, leading to increased graft loss and morbidity in some individuals. Treatment with interferon and other agents is controversial and not highly efficacious. The use of an effective antiviral agent to reduce or eliminate viral burden is desirable. To this end, we performed an open-label pilot trial to determine if rimantadine would show antiviral activity against hepatitis C virus (HCV) in the posttransplantation setting. Eleven patients with recurrent post-liver transplantation disease, characterized by transaminase level abnormality and HCV RNA in serum and liver biopsy specimens consistent with HCV infection were offered enrollment onto the study. Patients were treated for 12 weeks with rimantadine, 100 mg orally twice daily, and followed up after treatment for up to 8 additional weeks. Serum was collected at 2-week intervals to assess transaminase and HCV RNA levels. Nine patients completed the planned course of therapy. There was no significant change in serum alanine aminotransferase levels during treatment. No patients cleared HCV RNA from the serum, and fluctuations in the viral titer were not clearly associated with the initiation and completion of the active-treatment phase. Rimantadine was well tolerated, with only one patient who stopped therapy for perceived side effects. We conclude that rimantadine monotherapy has no role in the management of recurrent hepatitis C after liver transplantation. PMID- 9873089 TI - Etiology and outcome for 295 patients with acute liver failure in the United States. AB - Little information is available on acute liver failure (ALF) in the United States. We gathered demographic data retrospectively for a 2-year period from July 1994 to June 1996 on all cases of ALF from 13 hospitals (12 liver transplant centers). Data on the patients included age, hepatic coma grade on admission, presumed cause, transplantation, and outcome. Among 295 patients, 74 (25%) survived spontaneously, 121 (41%) underwent transplantation, and 99 (34%) died without undergoing transplantation. Ninety-two of 121 patients (76%) survived 1 year after transplantation. Acetaminophen overdose was the most frequent cause (60 patients; 20%), followed by cryptogenic/non A non B non C (NANBNC; 15%), idiosyncratic drug reactions (12%), hepatitis B (10%), and hepatitis A (7%). Spontaneous survival rates were highest for patients with acetaminophen overdose (57%) and hepatitis A (40%) and lowest for those with Wilson's disease (no survivors of 18 patients). The transplantation rate was highest for Wilson's disease (17 of 18 patients; 94%) and lowest for autoimmune hepatitis (29%) and acetaminophen overdose (12%). Age did not differ between survivors and nonsurvivors, perhaps reflecting a selection bias for patients transferred to liver transplant centers. Coma grade on admission was not a significant determinant of outcome, but showed a trend toward affecting both survival and transplantation rate. These findings on retrospectively studied patients from the United States differ from those previously gathered in the United Kingdom and France, highlighting the need for further study of trends in each country. PMID- 9873090 TI - Liver transplantation in patients with splenorenal shunts: intraoperative flow measurements to indicate shunt occlusion. AB - Two patients with previous distal splenorenal shunts (DSRSs) performed 6 years earlier underwent liver transplantation (LT). A preoperative selective mesenteric artery angiogram showed collateral veins draining mesenteric venous flow into the shunt. Intraoperative flow measurements were performed to assess the steal of portal venous flow by the shunt and determine the need for shunt occlusion. Portal vein, hepatic artery, and shunt flows were measured by ultrasound transit time flow probes in the native liver and after graft implantation with and without temporary shunt occlusion. Hemodynamic studies showed that long-standing DSRSs are high-flow shunts that steal portal flow. After graft implantation, DSRS flows remained high. Occlusion of the shunts produced an increase in portal vein flow at an amount similar to those of splenorenal shunt. Thus, the flow measurements showed persistent steal by the shunts after graft implantation and, therefore, the DSRSs were occluded but splenectomy was not performed. We conclude that the decision to occlude a DSRS should be based on the demonstration of steal of portal flow by the shunt and reversibility once the shunt is occluded. Splenectomy is not required when the DSRS is occluded. PMID- 9873091 TI - Nonspecificity of monoclonal antibody Tordji-22 for the detection of hepatitis C virus in liver transplant recipients with cholestatic hepatitis. AB - Detection of hepatitis C virus (HCV) antigens in liver tissue provides important diagnostic and pathological information. Limited studies have been performed on tissue taken after liver transplantation for HCV. In this study, serial post liver transplantation biopsy tissue from patients with recurrent HCV was tested, with particular interest in patients showing severe cholestatic hepatitis. HCV related antigens were detected using the commercial monoclonal antibody, Tordji 22. Initial results were promising, showing intense positive staining, especially in areas of hepatocyte ballooning. HCV-negative donor tissue was consistently negative by staining. However, as a final control for the level of tissue damage, HCV negative posttransplantation biopsy tissue showing hepatocyte ballooning was examined. These tissues also showed positive staining. All attempts to eliminate this nonspecific interaction failed. In conclusion, Tordji-22 was associated with nonspecificity in this posttransplantation population, and care is warranted when using this monoclonal antibody. PMID- 9873092 TI - Hepatocellular carcinoma after renal transplantation in the absence of cirrhosis or viral hepatitis: a case series. AB - The occurrence of hepatocellular carcinoma (HCC) in renal transplant recipients has typically been associated with hepatitis B or C infection. We encountered two cases of HCC in renal transplant recipients with negative hepatitis B and C markers and no underlying liver pathology, in whom immunosuppression therapy consisted of prednisone and azathioprine (AZA). Patient no. 1 is a 66-year-old man with diabetes who underwent cadaveric renal transplantation 13 years before presentation. An ultrasound obtained for evaluation of a prolonged prothrombin time and decreased serum albumin level showed a suspicious nodular lesion in the left lobe of the liver. A computed tomographic (CT) scan confirmed a 4- x 5- x 5 cm mass that, on biopsy, was determined to be well-differentiated HCC. There was no evidence of metastasis, and the results of random biopsies of the surrounding parenchyma were normal. The patient underwent a left lateral segmentectomy, did well, and an initial alpha-fetoprotein (AFP) level of 85995 ng/mL decreased to 9 ng/mL. Approximately 20 months postoperatively, however, a surveillance CT scan showed three hypervascular lesions in the right lobe of the liver and the AFP level increased to 28,370 ng/mL. Subsequent percutaneous alcohol injections yielded good results, and the patient is alive and well 13 months later. Patient no. 2 is a 57-year-old man who underwent cadaveric renal transplantation 24 years earlier. A CT scan of the abdomen obtained for evaluation of lower abdominal pain showed a 4- x 4- x 6.5-cm mass in the right lobe of the liver that, on biopsy, was found to be poorly differentiated HCC. Multiple biopsies of adjacent liver parenchyma showed no evidence of cirrhosis, AFP level was normal, and imaging studies showed no evidence of tumor spread. The patient underwent a right hepatic lobectomy and is doing well without evidence of recurrence 27 months postoperatively. Our two patients had no evidence of viral hepatitis, cirrhosis, or metabolic liver disease, yet both developed HCC. The use of AZA may have had a role in the development of HCC. In renal transplant recipients on long-term immunosuppression therapy, particularly AZA, it is prudent to maintain a high index of suspicion for HCC when liver enzyme level or function abnormalities are encountered. PMID- 9873093 TI - Hereditary hemochromatosis in liver transplantation. AB - A candidate gene, HFE, was recently described in patients with hereditary hemochromatosis (HH) and found to contain a missense mutation leading to a cysteine to tyrosine substitution (C282Y). A second mutation, H63D, was also found in the gene. This study was undertaken to determine the HFE genotype in liver transplant recipients clinically diagnosed with HH and those incidentally found to have increased iron deposition in their explanted livers and to evaluate whether biochemical or histological hepatic iron indices (HIIs) correlated with homozygosity for the C282Y mutation. We identified 15 patients clinically diagnosed with various liver disorders other than HH who had increased liver iron deposits among 918 adult patients who underwent liver transplantation from 1988 to 1995. Four additional patients were clinically diagnosed as having HH. Archival explant liver tissue was evaluated for the histological HII according to the method of Deugnier et al, in which an index greater than 0.15 suggests homozygosity for HH. The HII was computed according to established methods, with a value greater than 1.9 suggesting homozygosity for HH. A portion of liver tissue was subjected to DNA genotyping using polymerase chain reaction-amplified products. Two of 4 patients with clinically suspected HH were homozygous for C282Y, and 2 patients had neither mutation. One of the 15 patients not suspected to have HH was a C282Y homozygote, 1 was a C282Y heterozygote, 6 were H63D heterozygotes, and 7 had neither mutation. The histological HII was consistent with HH in 13 patients, whereas the HII was consistent with HH in 6 patients. Thus, in patients with end-stage liver disease, despite fulfilling the established clinical criteria for HH using biochemical and histological parameters, only a minority of patients were homozygous for the C282Y mutation. Hepatic iron overload may result from other causes, and in end-stage liver disease, an elevated HII may not accurately predict HH. Other factors that either control or lead to iron absorption may explain iron overload in these patients. PMID- 9873094 TI - Pulmonary vascular morphological changes in cirrhotic patients undergoing liver transplantation. AB - Cirrhosis and portal hypertension may be associated with pulmonary hypertension and pulmonary dysfunction. However, morphological pulmonary vascular lesions in patients with cirrhosis have not been well characterized morphometrically. We morphometrically evaluated pulmonary vessels in liver transplant recipients with pretransplantation cirrhosis and correlated our findings with pretransplantation cardiopulmonary function, postoperative course, and postmortem cardiopulmonary findings. Autopsy lung slides from 23 transplant recipients with pretransplantation cirrhosis were examined. External vessel diameter, intimal thickness, and arterial medial thickness were measured with a micrometer after pentachrome staining. The percent of total diameter comprised by intima or media was calculated for each vessel. Medical records were reviewed for smoking history, pretransplantation cardiopulmonary function testing, and postoperative course. Autopsy cases without liver or significant cardiopulmonary diseases, matched for age, sex, and smoking history, served as controls. Transplant recipients had significantly more pulmonary venous intimal thickening than matched controls (P <.0001). Sixty-five percent (15 of 23) of these patients had some degree of pretransplantation pulmonary dysfunction, defined by abnormalities in pulmonary function tests, oxygen saturation, and/or increased pulmonary artery pressures. However, the severity of venous intimal thickening did not correlate with the severity of pretransplantation pulmonary dysfunction. Arterial intimal and medial thickness were not statistically significantly different from controls. Pulmonary venous intimal thickening and resultant luminal impingement are morphological findings not previously described in this population. The arterial lesion, when present, is similar to that seen in pulmonary hypertension from other causes. These pulmonary vascular lesions may be implicated in pulmonary dysfunction in patients with cirrhosis and may be associated with increased posttransplantation cardiopulmonary morbidity and mortality. PMID- 9873095 TI - Management of hepatic metastases. AB - Although the liver is the most common site of metastatic disease from a variety of tumor types, isolated hepatic metastases most commonly occur from colorectal cancer and, less frequently, from neuroendocrine tumors, gastrointestinal sarcoma, ocular melanoma, and others. Complete evaluation of the extent of metastatic disease, both intrahepatically and extrahepatically, is important before considering treatment options. Based on a preponderance of uncontrolled studies for hepatic metastatic colorectal carcinoma, surgical resection offers the only potential for cure of selected patients with completely resected disease, with 5-year survival rates of 25% to 46%. Systemic and hepatic arterial infusion chemotherapy may be useful treatment options in patients with unresectable disease and possibly as an adjuvant treatment after liver resection. Other techniques of local tumor ablation, including cryotherapy and radiofrequency ablation, although promising, remain unproved. Management of hepatic metastases from neuroendocrine tumors and other noncolorectal primary tumors should be individualized based on the patient's clinical course, extent of disease, and symptoms. PMID- 9873096 TI - De novo hepatocellular carcinoma in a hepatic allograft with recurrent hepatitis C cirrhosis. AB - We report a case of de novo hepatocellular carcinoma (HCC) in a patient with recurrent hepatitis C (HCV) and cirrhosis 7 years after orthotopic liver transplantation (OLT). This is a previously unreported observation in the natural history of posttransplantantion HCV infection and reiterates the strong oncogenic potential of HCV. PMID- 9873097 TI - Inflammatory pseudotumor of the liver: a rare benign tumor mimicking a malignancy. AB - We describe a 9-month-old boy in whom a hepatic tumor called angioma was diagnosed on ultrasonography, performed for abdominal pain. He was asymptomatic until 9 months later, when he presented with weight loss, jaundice, and a hard tumor in the left liver lobe. Radiological examination showed a calcified and heterogeneous tumor, amputation of the left portal vein, and dilatation of bile ducts, strongly suggesting malignancy. However, liver biopsy showed the typical findings of inflammatory pseudotumor, which are myofibroblastic cells and perivascular plasmocytes in a dense collagenous stroma. The tumor was surgically removed without local recurrence with a 2-year follow-up. Inflammatory pseudotumor in childhood is more common in the lung and elsewhere is often mistaken for a slowly growing cancer. This case points out the difficulties in the radiological evaluation of liver tumors in childhood and the importance of the histological differentiation of this lesion from malignancy before laparotomy for adequate indication of the treatment, which includes no medical treatment and surgery as conservative as possible. PMID- 9873099 TI - Introduction PMID- 9873098 TI - Cause and prognosis in acute liver failure. PMID- 9873100 TI - Altered fractionation trials in head and neck cancer. AB - Advancement in radiobiological concepts has led to the development of two classes of new fractionation schedules for the treatment of head and neck cancers. These altered fractionation regimens are referred to as hyperfractionation and accelerated fractionation schedules. Hyperfractionation exploits the difference in fractionation sensitivity between tumors and normal tissues manifesting late morbidity. In contrast, accelerated fractionations attempt to reduce tumor proliferation as a major cause of radiotherapy failure. Although there are many permutations in accelerating radiation treatment, the existing schedules can be conceptually grouped into two categories, pure accelerated fractionations and hybrid accelerated regimens, depending on whether there are concurrent changes in other fractionation parameters. The results of 10 completed phase III clinical trials addressing different types of altered fractionation schedules in head and neck carcinomas are examined and summarized in this review article. The data of trials on hyperfractionation regimens applying 10% to 15% total dose increment over the standard 66 to 70 Gy consistently revealed a 10% to 15% improvement in the local control rate of a subset of intermediate-stage carcinomas without an appreciable increase in the incidence of late complications. The available data on accelerated fractionation regimens showed that tumor clonogen proliferation is a major cause of radiation failure. Completed studies, however, revealed that a 1 to 1.5-week treatment acceleration without total dose reduction achieved by administering 2-Gy fractions 6 times per week or concomitant boost schedule yielded an approximately 15% higher tumor control rate without increasing late toxicity. Shortening the overall time to less than 2 weeks with an associated total dose reduction did not seem to improve tumor control rate, with an exception perhaps for a small subset of patients with laryngeal carcinomas, but might decrease some late normal tissue injury. A weekly dose accumulation rate of 14 Gy or greater, or delivery of 3 fractions of 1.6 Gy/d, with 6 hour intervals, without total dose reduction, was found to increase morbidity beyond the acceptable level. Further treatment refinements building on these findings are being pursued. PMID- 9873101 TI - Radiotherapy and concurrent chemotherapy for the treatment of locally advanced head and neck squamous cell carcinoma. AB - Cure of locally advanced squamous cell carcinoma of the head and neck (SCCHN) is uncommon with radiotherapy alone. The desire for organ preservation in advanced resectable SCCHN and the need for better local therapy for unresectable disease have led to the development of treatment using radiotherapy and concurrent chemotherapy (RT/CCT). RT/CCT is an attractive strategy because the appropriate drug(s) may enhance radiation effects and independently contribute to local cytotoxicity. Concurrent treatment may combat tumor repopulation and provide the earliest possible treatment of distant micrometastases. RT/CCT may be integrated in synchronous or alternating schemes. Most randomized trials of RT/CCT versus radiation alone show superior local control, disease-free survival, and survival with combined modality treatment. Improved efficacy with RT/CCT is accompanied by increased acute toxicity, which necessitates compromises in the treatment design of most programs. Consequently the most effective RT/CCT regimen has not been defined. Chemical modifiers of toxicity are now under investigation in clinical trials and may allow for improved integration of RT/CCT. PMID- 9873102 TI - Combined radiotherapy and chemotherapy for nasopharyngeal carcinoma. AB - Among squamous cell carcinomas of the head and neck, nasopharyngeal carcinoma is probably the most radiosensitive and chemosensitive. It also has the highest incidence of distant metastasis. This article reviews the results of randomized trials of combined chemotherapy and radiotherapy for nasopharyngeal carcinoma to date. Induction chemotherapy with bleomycin, epirubicin, and cisplatin was shown to increase disease-free survival but not overall survival in a trial by the International Nasopharyngeal Cancer Study Group. Concurrent radiotherapy and cisplatin followed by adjuvant cisplatin and 5-fluorouracil infusion significantly decreased local, nodal, and distant failures and increased progression-free and overall survival in the Head and Neck Intergroup Trial. The toxicity of combined chemotherapy and radiotherapy, however, primarily acute toxicity, was significantly greater than that of radiotherapy alone. Further clinical trials using novel drugs, altered fractionation radiotherapy and chemotherapy dose schedules, new radiotherapy techniques, and other treatment modifiers are needed to further improve the therapeutic ratio. PMID- 9873103 TI - Larynx preservation trials: a critical appraisal. AB - Controlled trials testing the concept of laryngeal preservation for patients with locally advanced stage III and IV cancers of the larynx or hypopharynx were initiated in the mid-1980s. Three randomized trials, evaluating the same cisplatin plus 5-fluorouracil induction chemotherapy regimen and conventional radiotherapy compared with surgery and radiotherapy, have been completed, and the results of two are published. In addition, a meta-analysis of these three trials was completed. The studies are critically reviewed. Conclusions from these trials are that the induction approach is feasible; local and regional control are not improved, whereas distant metastases are delayed; there is no evidence of a difference in overall survival; and of the patients alive at 3 and 5 years, a functional larynx can be preserved in 67% and 58%; there are not enough data to know if there are differences in outcome by subsite. Several unanswered questions are being addressed in phase III trials. These include defining the precise contribution of chemotherapy by comparing induction chemotherapy and radiotherapy to treatment with radiotherapy alone; determining if elective neck dissection for patients with N2N3 neck disease would improve survival; and determining whether local control can be improved by using concomitant or alternating chemotherapy and radiotherapy. PMID- 9873104 TI - T3-4 squamous cell carcinoma of the larynx treated with radiation therapy alone. AB - This article reviews the results of radiation therapy alone for T3-4 squamous cell carcinoma of the larynx. Local control after radiation therapy is approximately 65% for patients with T3 glottic and supraglottic carcinoma. The likelihood of local control is inversely related to tumor volume and, for those with glottic cancers, is adversely influenced by cartilage sclerosis on pretreatment computed tomography of the larynx. Local control after radiotherapy is approximately 60% for T4 glottic cancers and 50% for T4 supraglottic carcinomas. The probability of severe complications is related to primary site and T-stage. PMID- 9873105 TI - The role of brachytherapy in head and neck cancer. AB - By precision dose delivery to a well-defined target, brachytherapy fulfills the complementary goals of delivering sufficient dose for tumor cell kill while sparing normal adjacent structures. The unique properties of this modality, either used alone or implemented in a combined modality setting with surgery or external-beam irradiation, offer an ideal means of enhancing the therapeutic ratio by exploiting the effect of local therapies while respecting normal tissue tolerance. Brachytherapy also provides an effective technique for the retreatment of patients with recurrent, persistent, or second primary head and neck malignant tumors in a previously irradiated region. The benefits of this treatment modality are reflected in excellent reported local control rates, shortened overall treatment times, reduced functional deficits, improved quality of life, and decreased costs. Successful brachytherapy relies, however, on close collaborative efforts between radiation oncologists and surgeons, careful assessment of the patient, precise planning, and technique that adheres to the rules of a system. Brachytherapy has been clearly identified as an effective, safe, and appealing modality in the treatment of head and neck malignancies. PMID- 9873106 TI - Toxicity antagonists in head and neck cancer. AB - Clinical trials of altered fractionation and concurrent chemoradiation regimens have better elucidated the limits of both acute and late normal tissue toxicities in the head and neck. Acute effects on mucosal epithelium represent the principal barrier to intensification of radiation or chemoradiation schedules. Late soft tissue injury and organ dysfunction limit efforts to escalate radiation total dose. New insights into the cellular and molecular mechanisms of injury repair allow new strategies in the management and prevention of treatment-related toxicity. Toxicity antagonists are agents that directly interfere with the mechanism of toxicity or modulate the normal tissue response to injury. This article reviews 10 agents under development. Not only could such interventions reduce treatment-related morbidity, but they may also allow treatment intensification in advanced disease. PMID- 9873107 TI - Chemoprevention in head and neck cancer: basic science and clinical application. AB - The concept of field cancerization and the multistep carcinogenesis theory are the premises on which the development of chemoprevention efforts has been based. The first concept underlies the fact that patients who have been cured of a cancer in the aerodigestive tract are still at risk of developing additional primary tumors in the same field, and the multistep carcinogenesis theory implies that the conceivable arrest of one of the steps by chemoprevention might be enough to impede the development of cancer. There is ongoing research for development of new candidate-chemopreventive substances in most areas of oncology. Most of the clinical experience in chemoprevention of head and neck cancer is based on the use of retinoids. Retinoids in high doses have demonstrated activity in treating oral premalignant lesions (OPLs) but with frequent side effects and early relapse after cessation of therapy. Subsequent trials showed better tolerability of retinoids in lower doses. Ongoing trials are currently evaluating whether chemoprevention over 3 years might have longer lasting effect both on OPLs and in the prevention of second primary tumors. PMID- 9873108 TI - Utilization of natural products for treatment of blood diseases. AB - This chapter presents an introduction to several diseases of the blood including infectious mononucleosis, leukemia, thrombosis and coagulation, bone marrow disorders, malaria, and anemia. In addition a survey of the recent literature is presented relative to natural products that have been utilized for the treatment of these diseases. The natural products that are reported represent a wide range of structural types and present interesting mechanisms of action. Thus the possibility exists that new drugs may be developed from these natural products which are more effective than those currently on the market. PMID- 9873109 TI - Beta-lactamase inhibitors: agents to overcome bacterial resistance. AB - The extensive use of beta-lactam antibiotics in hospitals and community has created major resistance problems leading to increased morbidity, mortality and health-care costs. Resistance is most often mediated by -lactamases, which have emerged in both gram-positive and gram-negative bacteria. A novel approach to countering bacterial beta-lactamases is the delivery of a beta-lactam antibiotic in combination with a beta-lactamase inhibitor. Several such combinations are currently available, containing inhibitors clavulanic acid, sulbactam and tazobactam. These inhibitors are not, however, active against all beta-lactamases and the AmpC chromosomal enzymes that are hyperproduced by some enterobacteria and pseudomonas are a particular gap. Moreover, genes for these AmpC enzymes have begun to escape to plasmids. Consequently, there is a growing need for new broad spectrum beta-lactamase inhibitors. This review offers an overview of synthetic beta-lactamase inhibitors, emphasizing information on their structures, and highlighting their activity against various beta-lactamases, particularly AmpC enzymes. Effective inhibition of AmpC enzymes are to be found among the penems and monobactams, but none of these has yet proved suitable for pharmaceutical development. PMID- 9873110 TI - Progress towards the discovery of orally active thrombin inhibitors. AB - The serine protease thrombin is a key enzyme in the control of blood coagulation and displays numerous other effects on platelet, endothelial and smooth muscle cell function. The pre-eminence of thrombin in the coagulation cascade has made the enzyme a popular drug target in the search for more clinically acceptable and safe anti-coagulants. This concept has been particularly strengthened by the demonstration that direct inhibitors of thrombin such as Revasc are clinically effective. A number of low molecular weight thrombin inhibitors have now been described in the literature although to date because of their inherent low bioavailability compounds have been limited to the parenteral route of administration. The introduction of appropriate pharmacokinetic properties into these first generation thrombin inhibitors has been problematic despite intensive research in this area. However, the first pre clinical examples of direct thrombin inhibitors possessing good oral bioavailability is now emerging. This review updates current developments in the progress towards the discovery of orally available thrombin inhibitors and suggests that the first clinical validation of drugs from this field is imminent. PMID- 9873111 TI - 1. The chemistry and pharmacology of indole-3-carbinol (indole-3-methanol) and 3 (methoxymethyl)indole. [Part II]. AB - Indole-3-carbinol (I3C) (2) is produced endogenously from naturally occurring glucosinolates contained in a wide variety of plant food substances including members of the family Cruciferae, and particularly members of the genus Brassica, whenever they are crushed or cooked. The acid environment of the gut very facilely converts it into a range of polyaromatic indolic compounds, e.g. (3, 4,5), which appear to be responsible for many of the physiological effects observed following the ingestion of these foods. These so-called chemopreventive compounds are important because of their enzyme induction and suppression, mutagenic, carcinogenic and, particularly, antimutagenic and anticarcinogenic properties against a variety of classes of carcinogens. These properties as well as other miscellaneous properties of these substances are critically reviewed in detail in this paper of >170 references, the second of two parts. At the present time it appears that I3C and its congeners have considerable potential as natural prophylactic anticancer agents against certain common neoplasms, especially inasmuch modern diets are increasingly deficient in these vegetable-derived substances. A short general assessment of the substantial potential of the title compounds concludes the review. PMID- 9873112 TI - Bioisosteric approach in the design of new dopaminergic/serotonergic ligands. AB - Dopaminergic and serotonergic ligands are widely applied in the therapy of some severe diseases in humans connected to the malfunctioning of the corresponding membrane receptors within the CNS. However, no pharmaceuticals of this type with an ideal therapeutic index have been synthesized so far and there is a constant need of producing new dopaminergic/serotonergic ligands with improved properties especially with regard to undesirable side effects expressed after a prolonged therapy. Dopaminergic/serotonergic ratio turned out to be important for a fine tuning of pharmacological profile of new ligands. Employing a bioisosteric approach, we have synthesized numerous quinoxalinediones, benztriazoles, benzimidazoles and 2-substituted benzimidazoles as potential dopaminergic and/or mixed dopaminergic/serotonergic compounds. With this purpose, benzimidazole and its derivatives were incorporated into phenylethylamine, 3- and 4-substituted phenylethylpiperidine, 1-substituted 4-arylpiperazine and semirigid 2 aminotetralin frame and the resulting ligands were checked for the binding affinity at the D1 and D2 dopamine and 5-HT1A serotonin receptors in radioligand binding assays in vitro. Synaptosomal membranes prepared from bovine caudate nuclei and hippocampi served as a source of the dopamine and serotonin receptors, respectively. [3H]SCH 23390 (D1 receptor-selective), [3H]spiperone (D2 receptor selective) and 8 OH [3H]DPAT (5-HT1A receptor-selective) were employed as radioligands in competition binding assays. Properties of substituents introduced into position 2 of benzimidazole ring, as well as the nature of the frame into which benzimidazole pharmacophore was incorporated have been shown to determine ligand binding affinity, mode of action and receptor preference, i.e. dopaminergic/serotonergic affinity ratio. Benzimidazolyl-2-thione and benztriazole derivatives were the most potent dopaminergic/serotonergic ligands. Molecular ab initio calculations of the electronic properties of pharmacophoric entities of the new ligands revealed different electron density distribution around the benzene ring in the active and inactive ligands. It can be assumed that this difference influences the properties of pi-pi interactions in a receptor-ligand complex. The results are discussed in comparison with the data of other authors working on similar topics. PMID- 9873113 TI - Pyrroloquinoline and pyridoacridine alkaloids from marine sources. AB - Marine organisms are a rich source for natural products. Pyrrolo[4,3, 2 de]quinolines and pyrido[4,3,2-mn]acridines are of major interest as metabolites in sponges and ascidians. Many of these compounds have generated interest both as challenging problems for structure elucidation and synthesis as well as for their cytotoxicities. The isolation, structure proof, biological activities, chemical properties and synthesis have attracted the attention of chemists, biologists and pharmacists. The principal structural feature of these alkaloids is the core of a planar iminoquinone moiety which can intercalate into DNA and cleave the DNA double helix or inhibit the action of topoisomerase II. Of the makaluvamines, makaluvamine F and A are the most cytotoxic to the HCT 116 cell line. The enhanced toxicity of the makaluvamines towards xrs-6 cells shows that all of the makaluvamines, except makaluvamine B, act like m-AMSA and etoposide in inhibiting topo iso merases via cleavable complex formation, or via the direct induction of DNA double-strand breaks. They are also amongst the most potent inhibitors of topoisomerase II. Both makaluvamine A and C can decrease tumor size in a solid human tumor model. Discorhabdin A and C in contrast are of high cytotoxicity, but they exhibit no inhibition of topoisomerase II. As representatives of the derivatives of pyrido[4,3,2-mn]acridine, cystodytins, kuanoniamines and diplamine are the most potent to inhibit HCT replication. Eilatin, as a 1,10-phenanthroline derivative, can form complexes with metal ions. It has been shown that these metal complexes can bind to DNA by intercalation. The new members of the pyrrolo[4,3,2-de]quinolines and pyrido[4,3, 2-mn]acridines, such as veiutamine, discorhabdin G, tsitsikammamines, epinartins, arnoamines as well as sagitol are reviewed. Some successful syntheses of pyrrolo[4,3,2-de]quinoline ring system and pyrido[4,3,2-mn]acridine ring system are also reviewed in this article. PMID- 9873114 TI - Advances in indolo[2,3-a]carbazole chemistry: design and synthesis of protein kinase C and topoisomerase I inhibitors. AB - Indolo[2,3-a]carbazoles, their pyrrolo[3,4-c]anellated variants and structurally closely related bisindolylmaleimides represent a biologically highly interesting class of natural compounds which are potential anticancer agents. According to the ongoing literature new and efficient synthetic methods yield a great variety of these compounds which have been reported in detail. The biological activities and the inhibitory activities against the target enzymes protein kinase C and topoisomerase I are also discussed including structure activity relationships. A molecular binding model of the protein kinase C inhibitors with the target enzyme at the atomic level is presented and supported by X-ray crystallographic structures and by molecular modelling studies. PMID- 9873115 TI - 5-Lipoxygenase: a target for antiinflammatory drugs revisited. AB - Arachidonate 5-lipoxygenase is the key enzyme in leukotriene biosynthesis and catalyzes the initial steps in the conversion of arachidonic acid to biologically active leukotrienes. Leukotrienes are considered as potent potent mediators of inflammatory and allergic reactions which are locally released by leukocytes and other 5-LO expressing cells and exert their effects via binding to specific membrane receptors and, as suggested recently, the nuclear receptor PPARa. Because of the proinflammatory profile of leukotrienes it was assumed that leukotriene biosynthesis inhibitors and leukotriene receptor antagonists have a therapeutical potential in a variety of inflammatory diseases. Clinical studies confirmed the therapeutic value of the antileukotriene therapy in asthma but the results with leukotriene biosynthesis inhibitors in psoriasis, arthritis and inflammatory bowel disease were more or less disappointing. This review summarizes the biochemistry of the 5-lipoxygenase pathway, the pharmacology of FLAP and 5 lipoxygenase inhibitors and discusses possible criteria for the development of these drugs. PMID- 9873116 TI - Prepubertal acne: clinical presentation, evaluation, and treatment. PMID- 9873117 TI - Management of adult acne and acne variants. PMID- 9873118 TI - Side effects of acne therapy and their management. PMID- 9873119 TI - Psychosocial aspects of acne vulgaris. PMID- 9873120 TI - Approaches to acne scarring: a review. PMID- 9873121 TI - The many faces of rosacea. PMID- 9873122 TI - Rosacea: epidemiology and pathogenesis. PMID- 9873123 TI - Differential diagnosis of a red face. PMID- 9873124 TI - Psychological aspects of rosacea. PMID- 9873125 TI - Drug therapy of rosacea: a problem-directed approach. PMID- 9873126 TI - Laser therapy in the management of rosacea. PMID- 9873127 TI - Angiogenesis: potential target for dermatologic intervention. PMID- 9873128 TI - [Obtaining myo-cardial perfusion images synchronized with the ECG (gated-SPEC) after injecting MIBI during exertion: comparative study with echocardiography in 95 patients for the assessment of left ventricular function]. AB - Technical innovation has recently resulted in the routine use of gated-SPECT in myocardial perfusion imaging. In the present work we compare estimates of left ventricular function (LV cavity) by gated-SPECT with those of conventional echocardiography in a group of 95 ischemic patients, 49 of whom had previous myocardial infarction. Kappa analysis showed correlation coefficients of 0.67 for global function in the whole group and of 0.68 in patients with MI, as well as of 0.56 in the anterior wall, and 0.55 in the inferior wall. With these data, we believe that the technique is useful and it adds to the conventional perfusion SPECT imaging. Also, it is conveniently validated against echocardiography in our hands. PMID- 9873129 TI - [Myocardial perfusion SPET in the detection of coronary artery disease in orthotopic heart transplantation]. AB - The incidence of atherosclerosis of the heart transplant in long term survivors is 38% at 5 years. In the present work, myocardial perfusion 201Tl-SPET was assessed as a non-invasive diagnostic method for the detection of postransplant coronary artery disease, as well as its efficiency with regard to other techniques. Twenty patients, aged (47 +/- 9) years old, who underwent heart transplantation at least 3 years earlier, were studied by 201Tl-SPET. Qualitative and quantitative analysis of the images were performed. It was found ischemia in 6 patients, 4 of them asymptomatics. In 5 of the 6 positive cases by SPET coronary stenosis was found by angiography. Kappa coefficient and percent of agreement were k = 0. 76 and Pe = 90%, respectively. There were no relationships among rejection crisis, sepsis by cytomegalovirus and coronary artery disease detected by using 201Tl-SPET (p > 0.05). The most relevant risk factors in the sample were hypertension and hyperlipidemia. Two patients died because of coronary artery disease. It was confirmed by necropsy findings. These results suggest that thallium-201 myocardial perfusion tomography could be useful to detect coronary artery disease in the transplanted heart. PMID- 9873130 TI - [Correlation between 99m Tc-tetrofosmin uptake and P-glycoprotein expression in non-small-cell lung cancer]. AB - We used thoracic SPECT to study the 99mTc-tetrofosmin (TF) uptake in patients with non-small-cell-lung-carcinoma (NSCLC). The results were compared with the percentage of P-glycoprotein (Pgp) found in flow cytometric analysis (FC) of samples of surgically-resected tumor tissue. MATERIAL AND METHODS: A total of 21 patients with NSCLC were studied by means 99mTC-TF and thoracic SPECT. Image analysis included the determination of the TF uptake rate in the lung mass with respect to that of healthy tissue of the contralateral lung. These rates were compared with the percentage of Pgp expression according to FC. FC analysis was also carried out in 16 samples of healthy lung tissue obtained from the patients. RESULTS: In healthy lung tissue, the mean Pgp expression according to FC was 4.58 +/- 1.87%. The cutoff value used to differentiate between Pgp positive and Pgp negative tumors was considered to be the mean plus two standard deviations (8.32). The Pgp-positive tumors (> 8.32%) presented significantly lower uptake levels (1.28 +/- 0.39) than the Pgp-negative lesions (1.66 +/- 0.33) (p = 0.029). CONCLUSIONS: There is a inverse correlation between the Pgp expression as determined by FC analysis and 99mTc-TF in NSCLC. Thus, this radiopharmaceutical provides rapid and non-invasive information on Pgp expression in these lesions. PMID- 9873131 TI - [Radionuclide bone scanning in patients with breast carcinoma. Study of lesions by stages, locations and diagnostic assessment]. AB - The aim of this study was to evaluate the diagnostic role of bone scan in patients with breast carcinoma. With this purpose, we have studied 591 patients (4 males). In 261 of them, we were able to establish the stage of disease: 33 in stage I, 137 in stage II, 71 in stage III y 20 in stage IV. Metastatic lesions were detected in 6% of patients in stage I, 8.8% in stage II, 12.7% in stage III y 50% in stage IV. Altogether, 71 from 591 patients (12%) had metastatic lesions. In 4 patients (5.3%) we obtained a false positive result. The bone lesions found in the scintigraphic studies were classified as not metastatic lesions in 236 patients (39.9%) but 26 of them were malignant (11%). Another 193 patients (32.6%) presented scintigraphic studies considered as equivocal for metastasis and 33 of them were also malignant (17%). In 150 patients scintigraphic studies did not present lesions during the follow up. In other 156 patients isolated lesions were detected and 24 of them (15.4%) were malignant. Only 4 <> lesions were detected and not any <> image was observed in this study. PMID- 9873132 TI - Unusual imaging of varicose veins after radiolabeling of autologous low-density lipoproteins. AB - LDL-radiolabeling with various isotopes (123I, 125I, 131I, 111In, 99mTc and 67Ga, among others) has been used for imaging the arterial wall, monitoring the LDL kinetics and in particular to assess vascular surface lining and foam cell content. From 286 patients studied with 111In-LDL scintigraphy, only 9 patients presented clinical varicosis. In 2 of these 9 patients a predominant imaging of varicose veins has been shown with scintigraphy, while in the other 7 patients no positive scintigraphic imaging could be detected. In vitro and in vivo perfusion experiments did not reveal an explanation for this unusual results. The reason for that positive LDL-uptake into varicose veins is unknown. These findings indicate that localized LDL-uptake has to be interpreted carefully in order to avoid potential false positive results. PMID- 9873133 TI - [Painful tibial swelling in a young patient. Findings of radiology and radionuclide scanning. Anatomopathological correlation]. AB - We report on the case of a young man with a longstanding tumor in the left proximal tibia, in whom we had the opportunity to correlate both radiologic (plain films plus MRI) and bone scintigraphic findings with those of biopsy and clinical follow-up. The final diagnosis was bone fibrous dysplasia. The correlation between noninvasive diagnostic findings is discussed, and the convenience of using such test combination is also stressed in order to endorse a noninvasive approach to such patients, given the low malignant potential, the exception being those patients in whom there is a definite fractures risk. PMID- 9873134 TI - [Radionuclide scan of the salivary glands]. PMID- 9873135 TI - Outer radius-wall thickness ratio, a postmortem quantitative histology in human coronary arteries. AB - Although the anatomy, histology and pathology of human coronary arteries have been studied extensively, little is known about the functional relationship between vessel radius and wall thickness. It is the purpose of this study to present detailed measurements and to describe this relationship covering the range from the feeding coronary artery to the arterioles. Human hearts of 10 adults less than 36 +/- 3 years old were investigated immediately postmortem. Ten cubic tissue blocks, measuring about 10 mm in length on each side, were dissected from the left ventricular wall. After fixation by immersion, 15-microm sections were prepared and outer and inner perimeters of 52 arterial segments were digitalized. Vessel radius and wall thickness were calculated and plotted to show their relationship over the whole range of vessel calibers. Outer vessel radii ranged from 100 to 3,000 microm and wall thickness from 80 to 800 microm. Plotting the outer vessel radius against the wall thickness, the data points were found to cluster around a straight line. A significant correlation between the two parameters was found (R2 = 0.79). This mathematical correlation and the good agreement of the presented results with data from other species indicate a common physiologic concept. PMID- 9873137 TI - A mechanism for anterior transposition of the anal fin and its appendicular support in the western mosquitofish, gambusia affinis affinis AB - The interosseal and suspensory ligaments of the axial and appendicular skeleton of the Western Mosquitofish, Gambusia affinis affinis were investigated in whole mounted late embryonic and adult female and male G. a. affinis stained with alcian blue and alizarin red S, cleared, and viewed using differential interference contrast. The interosseal and suspensory ligaments of late embryonic female G. a. affinis are reduced prior to sexual differentiation and continue reduced in adult females. However, in late embryonic male G. a. affinis the interosseal and suspensory ligaments are well developed prior to sexual differentiation and become robust in adult males. Treating late embryonic female G. a. affinis with either 30.0 or 40. 0 &mgr;g/ diet of 17alpha methyltestosterone led to the premature calcification of the hemal spine of the 13th vertebra. Treating late embryonic female G. a. affinis with 17alpha methyltestosterone did not lead to the remodeling of the hemal spines of the 14th 16th vertebrae. Thus, in all female G. a. affinis treated with 17alpha methyltestosterone, no anterior transposition of the anal fin and its appendicular support was observed. However, treating late embryonic male G. a. affinis with 30.0 &mgr;g/g diet of 17alpha-methyltestosterone led to the premature calcification of the hemal spine of the 13th vertebra, blocking the anterior transposition in only 62.5% of the males treated, and treating late embryonic male G. a. affinis with 40.0 &mgr;g/g diet of 17alpha methyltestosterone led to the premature calcification of the hemal spine of the 13th vertebra, blocking the anterior transposition in all (100.0%) males treated. Treatment with either 30.0 or 40.0 &mgr;g/g diet of 17alpha-methyltestosterone appeared not to completely effect the interosseal and suspensory ligaments of late embryonic male G.a. affinis as evident by the male-specific remodeling of the hemal spines of the 14th-16th vertebrae. The results of this study implicate the interosseal and suspensory ligaments in the male-specific remodeling of the axial and appendicular skeletal elements. Following the resorption of the 13th hemal spine, the interosseal and suspensory ligaments in late embryonic male G. a. affinis develop enough tension to cause directional growth, bending, and elongation of the 14th-16th hemal spines, thus providing a mechanism for the anterior transposition of the sexually dimorphic anal fin and its appendicular support. The data also support the need to revise the interosseal and suspensory ligament nomenclature by assigning the designation of 'female' and 'male' to these ligaments. PMID- 9873136 TI - Smooth muscle cell hypertrophy and hyperplasia in the partially obstructed gut of the rat: a quantitative evaluation. AB - Partial surgical stenosis of the gut induces smooth muscle cell hypertrophy and hyperplasia in the loops upstream from the obstruction in a few days. In the present study we report a quantitative evaluation of these phenomena in the circular smooth muscle layer of the small intestine of the rat 7 days after a subtotal stenosis. In the loops upstream from the obstruction, lumen diameter and muscle wall thickness were found to be increased in comparison with downstream tracts. Morphometrical analysis showed that cross-sectional profile areas of smooth muscle cells, within the circular layer of upstream loops, significantly increased in size. Moreover, smooth muscle underwent a marked cell hyperplasia; in fact, estimates of the number of smooth muscle cell nuclear profiles turned out to be from 2.0 to 3.8 times greater in upstream loops than in downstream loops. The relation between the degree of lumen dilatation and the degree of the increase of the circular muscle layer thickness and of hypertrophic and hyperplastic response is discussed. PMID- 9873138 TI - Immunohistochemical study of the innervation of the boundary area of the hard and soft palates of the rat. AB - The palatal mucosa plays an important role for patients using full dentures. The posterior ridge of the denture is designed to fit on the border between the hard and soft palates; accordingly, this boundary area is of importance when the outline of the denture is designed. In the present study, a rich supply of nerve fibers was found in the mucosa of the boundary area of the hard and soft palates of the rat. An immunocytochemical examination revealed nerve fibers containing protein gene product 9.5 (PGP 9.5), calcitonin gene-related peptide (CGRP), substance P (SP), pituitary adenylate cyclase-activating peptide, vasoactive intestinal polypeptide (VIP), C-terminal flanking peptide of neuropeptide Y (c PON), or nitric oxide synthase (NOS). Thin nerve fibers with PGP 9.5, CGRP, or SP penetrated into the epithelium, reaching beneath the cornified layer and terminated as free nerve endings. VIP-, c-PON- and NOS-containing nerve fibers were distributed in the connective tissue. Many of the VIP- and c-PON-containing nerve fibers were associated with blood vessels. In addition, nerve fibers containing PGP 9.5, CGRP, SP and c-PON were observed around, and penetrating into, the taste buds in the boundary area. PMID- 9873139 TI - Transient architectonic features in the basolateral amygdala of the human fetal brain. AB - The architectonical differentiation in the basolateral nuclei of the human fetal amygdala - with special reference to transient structures - was studied using series of relatively thick Nissl-stained sections. These architectonic features were correlated with the process of migration. Radial glial fibers providing the scaffold of migratory routes can reliably be marked with the aid of antivimentin. In the 5th gestational month a transient feature is conspicuous in the inferior portions of the basolateral nuclei bordering upon the ganglionic eminence (proliferative zone): columnar cell clusters, separated by cell-sparse septa, extend from the poliferative zone to the nuclei. The width of the cell columns vary considerably between the different nuclei. In vimentin immunopreparations fibers are found inside these cell columns. So they most probably reflect clustered migratory streams. Two months later, instead of this merging area between the ganglionic eminence and the amygdaloid nuclei a cell-free capsule envelopes the nuclei and clearly separates them from the ganglionic eminence. Changes in cytoarchitectonics are accompanied by a distinct rearrangement of radial glial fibers. A basket-like arrangement of the vimentin-immunoreactive fibers around the cell columns inside the cell sparse septa is found. Towards the end of pregnancy radial glial fibers gradually vanish. A comparison of Nissl and vimentin preparations reveals that transient architectonic characteristics as visible in relatively thick Nissl sections may be correlated with migrational routes. PMID- 9873140 TI - The incidence and dimensions of the retroarticular canal of the atlas vertebra. AB - The retroarticular canal has been implicated in compression of the vertebral artery, where it passes over the posterior arch of the atlas vertebra, during extreme rotational movements of the head and neck. The incidence of this retroarticular canal is not known in the South African population. The aim of the present study was to record the incidence and the dimensions of the retroarticular canal in South African white and black adults, aged 20-80 years. In a total of 1,354 atlas vertebrae, 9.8% of sides (264 left and/or right sides) were classified as having complete retroarticular canals, of which 11.7% (31 sides) were right-only, 24.6% (65 sides) left-only and 31.8% (84 left plus right sides) bilateral canals. This incidence did not increase with age and was lower in whites than blacks, with white males having the lowest and white and black females alike having the highest incidence of the canal. Measurements of the retroarticular canal showed that there was no difference between left and right sides. However, the superoinferior diameter was significantly less than the anteroposterior diameter, in all but the right canals in the white female subgroup. This difference in the dimensions of the retroarticular canal will decrease the cross-sectional area of the space available for the vertebral artery passing through it and may compromise blood flow in the vessel. PMID- 9873141 TI - Intrinsic physiological and pharmacological properties of central vestibular neurons. PMID- 9873142 TI - Vestibular compensation. PMID- 9873143 TI - Vestibular neuritis. PMID- 9873144 TI - Meniere's disease. PMID- 9873145 TI - Benign paroxysmal positioning vertigo. PMID- 9873146 TI - Drug therapy of nystagmus and saccadic intrusions. PMID- 9873147 TI - Nonpharmacological treatment of nystagmus. PMID- 9873148 TI - Objective response detection in the frequency domain: comparison of several q sample tests. AB - The objective assessment of the hearing threshold requires a suitable statistical test for response detection. For checking the q spectral lines of an auditory evoked potential spectrum in the frequency domain, a q-sample test seems to be more favourable compared to q applications of a one-sample test. The response detection performance of three q-sample tests (q-sample analogue of Watsons U2 test, q-sample uniform scores test and a modified q-sample uniform scores test) has been checked by extensive Monte Carlo simulations. To compare the performance of the three test statistics, sensitivity was calculated and receiver-operating characteristics were constructed from the probability density functions estimated by the Monte Carlo simulations. In addition, a comparison on the basis of real near-threshold auditory brainstem response data was carried out. The modified q sample uniform scores test appeared to be the most powerful test. Some aspects of the practical application of this test are discussed in this paper. PMID- 9873149 TI - Frequency-specific threshold determination with the CERAgram method: basic principle and retrospective evaluation of data. AB - A method for the objective evaluation of the hearing threshold using cortical evoked response audiometry was developed. The method results in a kind of objective audiogram, visualizing the significance of an auditory evoked potential (AEP) in a scheme similar to a conventional audiogram. In the present implementation of the method, four frequencies are tested quasi-simultaneously (500, 1000, 2000 and 3000 Hz; intensity steps of 5 dB). The significance of an evoked potential is assessed by means of the Rayleigh test, which is applied to the phase values derived from certain time windows of the single-trial epochs. A retrospective analysis of 1, 920 threshold estimations in 240 subjects suggested that the detection threshold (lowest stimulus intensity yielding a significant response) was, on the average, 7.5 dB above the electrophysiological threshold (intensity where the AEP amplitude vanishes). The grand-average amplitude intensity characteristic was approximated by the function a(1 - exp(-I/b)), with a = 6.25 microV, b = 22.3 dB and I representing the intensity (in decibels) relative to the electrophysiological threshold. PMID- 9873151 TI - Adaptive denoising and multiscale detection of the V wave in brainstem auditory evoked potentials. AB - This paper describes a wavelet-transform-based system for the V wave identification in brainstem auditory evoked potentials (BAEP). The system combines signal denoising and rule-based localization modules. The signal denoising module has the potential of effective noise reduction after signal averaging. It analyses adaptively the evolution of the wavelet transform maxima across scales. The singularities of the signal create wavelet maxima with different properties from those of the induced noise. A non-linear filtering process implemented with a neural network extracts out the noise-induced maxima. The filtered wavelet details are subsequently analysed by the rule-based localization module for the automatic identification of the V wave. In the first phase, it implements a set of statistical observations as well as heuristic criteria used by human experts in order to classify the IV-V complex. At the second phase, using a multiscale focusing algorithm, the IV and V waves are positioned on the BAEP signal. Our experiments revealed that the system provides accurate results even for signals exhibiting unclear IV-V complexes. PMID- 9873150 TI - Reliability of 80-Hz amplitude-modulation-following response detected by phase coherence. AB - The reliability and frequency specificity of the 80-Hz amplitude-modulation following response (80-Hz AMFR) during sleep detected by phase coherence as a measure of the hearing threshold was evaluated in 169 affected ears of 125 children with hearing impairment. The 80-Hz AMFR at a carrier frequency of 1000 Hz was monitored in all 169 ears and the auditory brainstem response (ABR) elicited by 1000-Hz tone pips was evaluated in 93 ears. Both responses were examined during sleep, and the thresholds were compared with the behavioral hearing threshold, which was determined by standard pure-tone audiometry or play audiometry. In 24 ears of 22 children with various patterns of audiogram, the 80 Hz AMFR was examined at different carrier frequencies, and the threshold pattern was compared with the pure-tone audiogram to investigate the frequency specificity of 80-Hz AMFR. The mean and standard deviation of the difference between the 80-Hz AMFR at a carrier frequency of 1000 Hz and pure-tone thresholds of 1000 Hz was 3.8 and 12.9 dB, and that between the ABR and pure-tone thresholds was 6.8 and 14.1 dB, respectively. The threshold patterns of 80-Hz AMFR clearly followed the corresponding audiogram patterns in all types of hearing impairment. The measurement of 80-Hz AMFR thus appears to be accurate in hearing assessment and to have good frequency specificity in children during sleep. PMID- 9873152 TI - Modulations of early and late secretory processes by activation of protein kinases in the rat adrenal medulla. AB - Modulatory effects of the activation of either protein kinase C (PKC) by phorbol 12,13-dibutyrate (PDBu) or protein kinase A (PKA) by forskolin on stimulant evoked secretory processes in the perfused rat adrenal medulla were studied. PDBu or forskolin was applied during repetitive stimulation (30 s each at 10-min intervals) with nicotine, bradykinin, muscarine or histamine, and changes in [Ca2+]i (fura-2 microfluorometry) and catecholamine secretions (electrochemical detection) were simultaneously measured. PDBu markedly potentiated the nicotine evoked secretion without altering the [Ca2+]i response. PDBu partially inhibited the muscarine-evoked secretion and almost completely blocked the histamine-evoked secretion, concomitantly with extensive suppressions of the [Ca2+]i responses to these stimulants. The bradykinin-evoked secretion was enhanced by PDBu despite a slight attenuation of the [Ca2+]i response. PDBu reduced bradykinin-induced intracellular Ca2+ release in a Ca2+-free medium but enhanced the secretion associated with the released Ca2+. These results suggest that PDBu-activated PKC modulates secretory processes at, at least, two different stages. An early-stage modulation may downregulate receptor/G protein systems, which accounts for the inhibitory effect of PDBu on the muscarine- and histamine-evoked responses. A late-stage modulation may generally promote Ca2+-triggered exocytosis after elevation of [Ca2+]i, which explains the potentiation of the nicotine-evoked secretion by PDBu. The late-stage modulation may counteract the early-stage modulation in bradykinin-stimulated cells. Forskolin potentiated the secretory responses to the four secretagogues without increasing the [Ca2+]i responses. PKA may modulate secretory process at a step(s) distal to the rise in [Ca2+]i as is the case with the late-stage modulation by PKC. PMID- 9873153 TI - Stimulation of HCO3- secretion across cystic fibrosis pancreatic duct cells by extracellular ATP. AB - Previous studies have demonstrated that extracellular ATP is able to activate Ca2+-dependent Cl- channels which may be important in circumventing the defective cAMP-dependent pancreatic ductal HCO3- secretion in cystic fibrosis (CF). The present study further investigated the effect of extracellular ATP on the stimulation of HCO3- secretion across CF pancreatic duct cells (CFPAC-1). Cells were grown in culture plate inserts which enabled the access of a pH microelectrode to the apical compartment. Changes in apical pH upon stimulation by extracellular ATP, as an indication of HCO3- secretion, were measured. ATP induced a rapid increase in apical pH, which reached a plateau with an averaged pH unit of 0.3 higher than that measured in unstimulated cells. The effect of ATP was concentration dependent. The ATP-induced change in pH could be blocked by apical addition of Cl- channel blockers, indicating that activation of apical Cl- channel is vital for HCO3- secretion by the pancreatic duct cells. Together with the previous finding, the present study suggests that HCO3- secretion may be stimulated in CF pancreatic duct cells by extracellular ATP via a cAMP independent pathway. PMID- 9873154 TI - Vasopressin induces dopamine release and cyclic AMP efflux from the brain of water-deprived rats: inhibitory effect of vasopressin V2 receptor-mediated phosphorylation. AB - The neurohypophyseal hormone vasopressin (AVP) is widely distributed throughout the central nervous system. It acts as an excitatory transmitter in the CNS and plays an important physiological role in water and electrolyte homeostasis. However, water deprivation has been shown to induce changes in the levels of monoamines, but there is little knowledge about the influence of AVP on monoamine levels after water deprivation. In this study, we investigated the effect of AVP and its receptor antagonists on alterations in dopamine (DA) release and cyclic adenosine 3',5' monophosphate (cAMP) efflux from rat brain slices following water deprivation. Striatal brain slices (500 microm thick) were incubated in a medium with or without AVP (0. 1-1.0 microM) for 30 min. After 2 h of washout in normal medium, high KCl (40 mM)-evoked DA release and cAMP efflux from the rat brain slices were examined. In the brain slices of euhydrated animals, treatment with AVP slightly altered DA release and cAMP efflux from the brain. This increase in DA release and cAMP efflux was not significantly affected by the addition of a calcium/calmodulin-dependent protein phosphatase, calcineurin (20 microM), to the incubation medium or either by a V1 or V2 AVP receptor antagonist. In contrast, AVP significantly increased the DA release and enhanced the cAMP efflux from the brain slices of water-deprived animals. The AVP-induced increase of brain response in the water-deprived animals was significantly attenuated by a V2 receptor antagonist, partially by calcineurin, but not by a V1 receptor antagonist. The present results suggest that AVP may play a role in water deprivation-induced DA release and cAMP efflux, which is possibly mediated through the activation of the V2 receptor. The V2 receptor action is attenuated by calcium/calmodulin-dependent dephosphorlyation of some cellular proteins critical for signal transduction. PMID- 9873155 TI - Chronic exposure to hypergravity affects thyrotropin-releasing hormone levels in rat brainstem and cerebellum. AB - In studies to determine the neurochemical mechanisms underlying adaptation to altered gravity we have investigated changes in neuropeptide levels in brainstem, cerebellum, hypothalamus, striatum, hippocampus, and cerebral cortex by radioimmunoassay. Fourteen days of hypergravity (hyperG) exposure resulted in significant increases in thyrotropin-releasing hormone (TRH) content of brainstem and cerebellum, but no changes in levels of other neuropeptides (beta-endorphin, cholecystokinin, met-enkephalin, somatostatin, and substance P) examined in these areas were found, nor were TRH levels significantly changed in any other brain regions investigated. The increase in TRH in brainstem and cerebellum was not seen in animals exposed only to the rotational component of centrifugation, suggesting that this increase was elicited by the alteration in the gravitational environment. The only other neuropeptide affected by chronic hyperG exposure was met-enkephalin, which was significantly decreased in the cerebral cortex. However, this alteration in met-enkephalin was found in both hyperG and rotation control animals and thus may be due to the rotational rather than the hyperG component of centrifugation. Thus it does not appear as if there is a generalized neuropeptide response to chronic hyperG following 2 weeks of exposure. Rather, there is an increase only of TRH and that occurs only in areas of the brain known to be heavily involved with vestibular inputs and motor control (both voluntary and autonomic). These results suggest that TRH may play a role in adaptation to altered gravity as it does in adaptation to altered vestibular input following labyrinthectomy, and in cerebellar and vestibular control of locomotion, as seen in studies of ataxia. PMID- 9873156 TI - Photomodulation of the melanocyte cell cycle by indoleamines. AB - Melanocytes are photoresponsive cells which respond to varying doses of UV exposure in the G2 phase of the cell cycle by prominent dendricity. This photoresponse is related to indoleamine light sensitivity. The present study highlights the role of indoleamines in the photomodulation of the melanocyte cell cycle. The study was conducted on 40 whole-skin organ cultures taken from the marginal zone of vitiligo. Twenty organ cultures were subjected to G2-phase arrest, while 20 were incubated in tryptamine. The organ cultures were incubated in the dark, exposed to a pulse of 120 s UV at 2 h of incubation and harvested 3 and 6 h after UV exposure. It has been reported that the photosensitive enzymes N acetyl transferase (NAT) and hydroxyindole-o-methyl transferase (HIOMT) are activated during the G2 phase. The conversion of serotonin to melatonin is inhibited by UV exposure as seen at 3 h. This activity recovers on continued dark incubation 6 h after UV exposure. On incubation with tryptamine, UV exposure results in utilisation of tryptamine as seen by prominent indoleamine positivity. Three hours after UV exposure, there is 75% dendricity indicating G2 phase traverse. There is a corresponding high serotonin positivity with a low melatonin positivity. This is reversed following 6 h of dark incubation with high melatonin positivity indicating reactivation of NAT and HIOMT. This is accompanied by a doubling of the melanocyte number due to mitotic traverse and an arrest in G1 phase with low utilisation of tryptamine. Thus tryptamine is utilised by melanocytes on UV exposure to be synchronised and traversed into G2 phase activating the photosensitive enzymes NAT and HIOMT. When followed by a dark phase, melatonin accumulates to traverse the melanocytes through M-phase of the cell cycle with doubling of the cell number. Thus the uptake and metabolisation of indoleamine precursors photomodulate the melanocyte cell cycle on UV exposure. PMID- 9873157 TI - Strokes in the elderly - higher acute and 3-month mortality - an explanation. AB - AIM: Stroke is common in older people. The objective of the study was to determine if older stroke patients have a higher mortality and disability compared with younger patients for comparable stroke severity and pathology and whether there is an explanation for the difference. METHODS: A prospective study was undertaken in 296 consecutive patients admitted with acute stroke. Patients were studied for neurological features, pre-stroke functional disability, severity of stroke defined by stroke syndromes and pathology of stroke on CT scans (202 patients). Post-stroke disability was defined according to the functional status within 72 h of admission. A record was made of the intercurrent illness while the patients were in acute wards and of the risk factors. Patients were dichotomized into two age groups: younger group - up to 75 years (163 patients) and older group - over 75 years (133 patients). Outcome was measured according to (1) discharge status from acute wards, i.e., dead or alive, and (2) mortality at 3 months. RESULTS: Although there was no significant difference in severe clinical stroke syndromes (p = 0.72), CT scan features (p = 0.68) and pyrexia (0.38) between the two age groups, the older patients had significantly more disabling strokes as defined on Barthel Index (p = 0.015) and a higher mortality in the acute phase (p < 0.01) and at 3 months (p = 0.001). The older stroke patients had more severe pre-stroke disability (p < 0.001) and more severe neurological impairment for similar stroke severity and pathology. Early mortality was more influenced by pre-stroke global health than age whereas 3 month mortality was influenced by age to the exclusion of all other known prognostic factors. CONCLUSION: The older stroke patients have more disabling stroke and an increased mortality for a similar spectrum of stroke severity and pathology. The explanation for higher mortality of the older patients is the poor pre-stroke health and higher immediate post-stroke disability. PMID- 9873158 TI - Influence of clinical factors, CT findings and early management on outcome in supratentorial intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Treatment of supratentorial intracerebral hemorrhage (SICH) is still controversial and new adequately sized randomized controlled trials (RCTs) of surgical evacuation are greatly needed. Our aim was to identify and quantify the most important clinical and CT factors related to prognosis in patients with SICH, to estimate the treatment effect in various subpopulations of surgically treated patients and to make assumptions on target population and sample size in future trials. METHODS: Uni- and multivariate analysis of retrospectively collected data on clinical and CT factors on admission and early management in 203 patients with SICH, mortality at discharge, 30 days, 6 and 12 months and clinical outcome according to the modified Rankin Scale (mRS) at 6 months and follow-up at a mean of 3.1 years after admission. RESULTS: Level of consciousness according to the Glasgow Coma Scale (GCS) and age were the single two factors best related to mortality at 6 and 12 months. GCS and age, in association with hematoma volume and location, arterial hypertension and to some extent use of steroids, were also related to clinical outcome according to the mRS at 6 months and 3.1 years. Surgical evacuation seemed to have a positive effect on clinical outcome in only a small subgroup of the patients. CONCLUSIONS: Our data support a future RCT of surgical evacuation versus conservative treatment in SICH restricted to patients younger than 60-65 years with a GCS on admission in the range of 6-11 and a hematoma not mainly located in the thalamus with a volume in the range of 30-100 ml causing a midline shift of less than 10 mm. Randomization should be balanced within groups of patients with lobar and basal ganglion hematomas, arterial hypertension and intraventricular hemorrhage, and the use of steroids should be discouraged. PMID- 9873159 TI - Aggravation of acute ischemic stroke by hyperthermia is related to an excitotoxic mechanism. AB - OBJECTIVE: To examine whether hyperthermia aggravates cerebral injury in acute ischemia by an excitotoxic mechanism, we studied the relationship between body temperature on admission and CSF concentrations of neuroexcitatory amino acids in 128 patients with acute ischemic stroke of less than 24 h duration. METHODS: Stroke worsening was defined as the percent change between the Canadian Stroke Scale (CSS) at 48 h and the CSS on admission. Infarct volume was measured on days 4-7 on cranial computed tomography. Excitatory amino acids were analyzed using HPLC. RESULTS: Glutamate concentration [median (min.-max.)] was 11 (2-19) micromol/l in hyperthermic patients (body temperature >37.5 degreesC) and 5 (2 22) micromol/l in normothermic patients (p < 0.0001). Glycine concentration in hyperthermic and normothermic patients was 16 (3-21) micromol/l and 9 (3-50) micromol/l, respectively (p < 0.0001). Glutamate was significantly higher in patients with hyperthermia only during the first 12 h after the onset of symptoms. The CSF concentrations of glutamate (r = 0.52; p < 0.0001) and glycine (r = 0.62; p < 0.0001) correlated with body temperature. Body temperature was significantly related to stroke worsening and infarct size, but this effect was dependent on the glutamate effect. CONCLUSION: Glutamate and glycine release during the acute phase of cerebral ischemia could be responsible for the increased brain damage in hyperthermia. PMID- 9873160 TI - Thrombus in vertebrobasilar dolichoectatic artery treated with intravenous urokinase. AB - BACKGROUND: Vertebrobasilar dolichoectasia is often found in patients with posterior circulation ischemia. Brain ischemia is caused by abnormal flow in the dilated artery and obstruction of paramedian arteries or intraluminal thrombus with artery-to-artery embolism. We report a patient with vertebrobasilar dolichoectasia and luminal thrombus treated with intravenous urokinase who did well but died 2 months later of subarachnoid hemorrhage. CASE DESCRIPTION: A 60 year-old man developed right-hand clumsiness, dysarthria and ataxia. Computed tomography showed vertebrobasilar dolichoectasia and thrombus in the basilar artery. Symptoms quickly resolved on heparin but recurred on warfarin and again resolved on heparin. Two weeks later, while on warfarin and aspirin 325 mg, he developed hand numbness, oscillopsia and ataxia. Symptoms again resolved on heparin. Angiography showed severe dolichoectasia of the distal right vertebral artery and basilar artery. A large mural thrombus was detected in the ventral part of the distal basilar artery narrowing the lumen by 50%. He was treated with intravenous urokinase 4,400 units/kg as a bolus followed by 4,400 units/kg/h for 12 h. Repeat angiography showed almost complete recanalization and improved filling of basilar artery branches. He was maintained on warfarin and aspirin 81 mg and had no further ischemic episodes. He died 2 months later of rupture of the basilar artery and subarachnoid hemorrhage. CONCLUSION: Some patients with thrombosis of vertebrobasilar dolichoectactic arteries continue to have ischemic symptoms despite adequate anticoagulation. Intravenous thrombolysis may be effective in reducing the risk of stroke, but the risk/benefit ratio needs to be assessed in each patient. PMID- 9873161 TI - Cobalt-55 positron emission tomography of ipsilateral thalamic and crossed cerebellar hypometabolism after supratentorial ischaemic stroke. AB - Cobalt-55 (55Co) is a positron emission tomography (PET) tracer used to demonstrate brain damage, possibly associated to calcium-mediated processes. The degree of 55Co accumulation correlates with the severity of ischaemia in stroke patients. It is still a matter of debate whether ipsilateral thalamic hypometabolism (ITH) and crossed cerebellar hypometabolism (CCH), occurring after middle cerebral artery (MCA) infarcts, represent only a metabolic depression of these remote regions or can lead to structural damage. The present study investigates whether an increased 55Co influx can be demonstrated with PET in ITH and CCH after MCA infarcts. About half of the patients with ITH and CCH had a significant degree of 55Co uptake in, respectively, the ipsilateral thalamus and the contralateral cerebellar hemisphere. It was observed in patients with severe signs of stroke on admission and poor clinical outcome, and correlated well with the degree of 55Co influx within the supratentorial infarct. The present study demonstrates that ITH and CCH after MCA infarction can represent structural damage in these remote areas that occurs during the second week after stroke onset. PMID- 9873162 TI - Cardiac papillary fibroelastoma: aA rare cause of ischemic stroke in the young. AB - Among etiologies of stroke in young adults, primary cardiac tumors are very rare. We report the case of a 37-year-old woman who was admitted for an ischemic stroke in the right middle cerebral artery region. Etiologic investigations revealed, after transthoracic and transesophageal echocardiography, an aortic valve tumor. Treatment was first medical with anticoagulation, then surgical. Histological examination showed a papillary fibroelastoma. After a review of the literature, the possible mechanisms of the ischemic event are discussed and lead to the conclusion that this tumor must be surgically excised, even if asymptomatic, because of recurrent ischemic complications responsible for myocardial infarction, stroke and sudden death. PMID- 9873163 TI - True fungal mycotic aneurysm of the basilar artery: a clinical and surgical dilemma. AB - Mycotic aneurysms of the intracranial circulation of true fungal etiology are extremely rare and are associated with a very high mortality. We report a case of a fatal aneurysm of the basilar artery secondary to Scedosporium apiospermum infection. The medical and surgical treatments are presented to demonstrate the difficulties associated with these lesions. PMID- 9873164 TI - XV Congress of the Italian Society of Neurosonology, L'Aquila, October 1-4, 1998. PMID- 9873165 TI - Immunopathogenesis of psoriasis: news in an old concept. AB - Twenty years ago, a concept of psoriasis pathogenesis was proposed in which epidermal proliferation of psoriatic lesions was the consequence of immunological abnormalities. This concept has subsequently been supported by the remarkable efficacy of immunosuppressive drugs. Moreover, recent experimental data indicate that activated psoriatic lymphocytes are capable of inducing keratinocyte proliferation. However, we have still to explain the mechanism by which lymphocytes act on keratinocytes and to find out what antigenic material could be responsible for their activation. PMID- 9873166 TI - Electrical impedance of nodular basal cell carcinoma: a pilot study. AB - BACKGROUND: Previously, we have explored the use of measurements of electrical impedance and devised 4 physically distinct indices named magnitude index (MIX), phase index, real part index and imaginary part index (IMIX) from the impedance data. Our results indicated that these indices could characterize contact reactions. OBJECTIVE: The goal of the present study was to use the electrical impedance method for the preoperative assessment of nodular basal cell carcinoma (BCC). METHODS: We included 11 patients with a total of 12 nodular BCC, diagnosed clinically and histologically. The noninvasive measurements were performed by transepidermal water loss (TEWL) and electrical impedance. For reference, normal looking contralateral or ipsilateral skin was used. RESULTS: Compared to controls, the mean TEWL of BCC was increased, but this finding was not statistically significant. The electrical impedance measurements of BCC tissue revealed statistically significant changes of the impedance indices MIX and IMIX (p /=10.3 pmol/l and group 2 (n = 15) with FT4 <10.3 pmol/l. TSH was measured by immunoradiometric assay. TSH nadir, TSH peak and TSH surge were calculated. Both groups differed significantly from control group in TSH surge values: group 1 (p < 0. 05), group 2 (p < 0.01). TRH test was abnormal in 11/27 patients of group 1 and 10/15 patients of group 2. In group 1, 7 patients had normal tests, 2 had abnormalities in both tests, 9 had only TSH nocturnal surge altered and 9 showed only TRH alterations. All patients of group 2 presented thyroid axis abnormalities. In conclusion, in patients with hypothalamic pituitary disorders with low FT4, no further investigation is required to demonstrate thyroid axis alterations, however in patients with normal FT4, nocturnal TSH secretion and TRH test may be required to evidence thyroid abnormalities. PMID- 9873193 TI - Developmental gene expression and immunohistochemical study of the human endocrine pancreas during fetal life. AB - The development of human endocrine pancreas has been the subject of many immunohistochemical studies but very little is known at the molecular level. We have determined the patterns of gene expression of glucagon, somatostatin and pancreatic polypeptide during fetal life (16-41 weeks of gestation) using the dot blot hybridization method. In spite of some dispersion in the mRNA levels, no progressive increase or decrease during this period of fetal life could be found, as previously observed for insulin. In keeping with these molecular data, no increase in immunostaining of the four hormones was observed, but a dispersion of endocrine cells within the exocrine tissue was noticed at 20 weeks of gestation followed by a clear differentiation of the Langerhans islets at 31 weeks. Interestingly, the mRNA levels of the four hormones were always higher in the fetal pancreas than in the adult pancreas. PMID- 9873194 TI - Catch-up growth after childhood-onset substitution in primary hypothyroidism: is it a guide towards optimal growth hormone treatment in idiopathic growth hormone deficiency? AB - Catch-up growth was analyzed in 20 prepubertal children with primary hypothyroidism (PH) starting treatment at an age of 4.4 (1.2-10.1) years and a height (HT) SD score (HT SDS) of -3.1 (+/-0.8). All patients were followed for at least 3 prepubertal years. HT velocity was 12.3 +/- 2.3, 9.0 +/- 1.8 and 7.5 +/- 2.2 cm/year, and change in HT SDS was 1.60 +/- 0.56, 0.57 +/- 0.33 and 0.28 +/- 0.38 during the 1st, 2nd and 3rd year, respectively. The 11 children followed to adult height reached a HT SDS of -0.11 +/- 1.1, all within their target HT range. HT gain (DeltaHT SDS) during the 1st year was correlated with the degree of catch up growth (r2 = 0.78, p < 0.001). While catch-up growth in childhood-onset PH is complete, this is not the case in GH deficiency (GHD). Based on the auxological characteristics of the patients with PH, HT velocities during the first 2 years were predicted applying prediction models devised for prepubertal children with idiopathic GHD. The modalities of GH treatment observed in the models were used to calculate predicted HT velocities of the PH patients. Observed HT velocities in PH were higher than predicted HT velocities during the 1st (10.67 +/- 1.37 cm/year, p < 0.01) and 2nd (8.35 +/- 0.86 cm/year, p = 0.128) year. The data show that catch-up potential in idiopathic GHD of childhood onset is reduced compared to PH. Since early catch-up as well as total HT recovery in children with GHD are often not reached by present treatment modalities, catch-up growth in PH may serve as a model towards optimizing GH treatment. The data suggest that initial GH doses of 1.0 IU/kg/week, rather than the presently recommended 0. 6 IU/kg/week, need to be given in GHD in order to achieve the degree of early catch up observed in PH and to consequently improve the final outcome. PMID- 9873195 TI - Study of serum leptin in cafeteria-diet-overfed rats. Influence of diet, insulin and corticosterone. AB - In a group of 15 male Wistar rats overfed with cafeteria foods (delivering a mean fat percentage of 60%) during 5 months from the age of 8 weeks and in a control group of 15 rats fed with a standard chow for the same period, serum leptin, insulin and corticosterone were measured by RIA and body composition was determined by dual-energy X-ray absorptiometry. Significantly higher fasting serum concentrations of leptin, insulin and corticosterone were found in the cafeteria-diet group. Fasting leptin concentrations were significantly higher in rats with a body fat percentage of more than 25% compared to the others, irrespective of the type of feeding. The log serum leptin correlated positively with body fat percentage and fasting insulin concentration but not with corticosterone concentration. Leptin concentration corrected for body fat mass was, however, comparable between the two diet groups, while the leptin/insulin ratio was lower in the cafeteria-diet group. In conclusion, chronic overfeeding resulting in an increased body fat percentage in rats is associated with hyperleptinemia, hyperinsulinemia and hypercorticism. Serum leptin levels appear to primarily track total body fat percentage and are unaffected by dietary fat manipulation in cafeteria-diet-induced obese rats. PMID- 9873196 TI - Longitudinal analysis of maternal serum leptin levels during pregnancy, at birth and up to six weeks after birth: relation to body mass index, skinfolds, sex steroids and umbilical cord blood leptin levels. AB - Leptin is an important regulator of body fat mass and energy expenditure during adult life. The mechanisms by which maternal and fetal weight are regulated during pregnancy are poorly understood. In order to gain more insight into a potential role of leptin during gestation, a prospective, longitudinal study was carried out to measure leptin concentrations in maternal serum of 29 healthy women during pregnancy up to 6 weeks after birth and also in umbilical cord blood of their newborns. Leptin concentrations were measured using a specific RIA. In addition, estradiol, testosterone, and sex hormone binding globulin were determined using commercially available RIAs. The mothers' skinfolds were determined at four sites using a Holtain caliper. Leptin levels increased continuously during pregnancy and reached 25.8 +/- 14.7 ng/ml at 38-40 weeks. At birth, leptin concentrations were 23.5 +/- 15.4 ng/ml. Three days after delivery a significant decrease of leptin levels to 10.6 +/- 6.0 ng/ml was observed. Six weeks after birth the leptin concentration in maternal serum was 13.8 +/- 8.6 ng/ml. At birth, maternal serum levels were significantly higher than levels in cord blood and did not correlate with leptin levels in cord blood or neonatal weight. Furthermore, leptin levels did not correlate with maternal sex steroids and sex hormone binding globulin levels. At 6-8 weeks of pregnancy, maternal leptin serum levels correlated significantly with BMI (r = 0.81). The correlation coefficients (leptin vs. BMI) dropped with increasing gestational age and at birth only a poor correlation persisted (r = 0.50). Six weeks after birth there was again a high correlation between leptin levels in maternal serum and BMI (r = 0.76). Subscapular skinfold thickness was correlated to leptin concentrations in maternal serum during the whole period of the investigation. In conclusion, maternal leptin levels continuously increased from 6-8 weeks up to 38-40 weeks of pregnancy. Maternal leptin levels decreased dramatically after birth. Six weeks after delivery, leptin levels were comparable to the values measured at the beginning of pregnancy. We hypothesize that leptin might play an important role during pregnancy and fetal development. PMID- 9873197 TI - Different effects of continuous and intermittent patterns of growth hormone administration on lipoprotein levels in growth hormone-deficient patients. AB - BACKGROUND: Lipoprotein (a) (Lp(a)) is a risk marker for the development of atherosclerotic coronary heart disease. Growth hormone (GH) administration to GH deficient (GHD) adults increases serum Lp(a) concentrations, and the levels of Lp(a) and GH are correlated in patients with acromegaly. Studies in rats have demonstrated differential effects of constant and intermittent GH patterns on levels of certain lipoproteins. The aim of the present studies was to describe the impact of intermittent and continuous patterns of GH delivery to GHD patients on serum levels of Lp(a) and other lipoproteins. METHODS: In one study (A) 10 GHD patients received in random order a fixed GH dose intravenously as: (1) continuous infusion; (2) eight bolus injections, and (3) a combination of 1 and 2. Each study lasted 36 h and was preceded by at least 4 weeks without GH. In another study (B) 13 GHD patients received GH in random order as: (1) continuous subcutaneous (s.c.) infusion, and (2) daily s.c. injections in the evening for 1 month each. The patients were studied during steady-state conditions at the end of each treatment period. RESULTS: In study A Lp(a) levels increased significantly following continuous (p < 0.05) and combined patterns (p < 0.02) of GH administration to GH-deprived GHD patients, whereas the increase after GH bolus injections alone was not significant (p = 0.14). In study B significantly higher (p < 0.05) serum levels of Lp(a) were obtained after continuous s.c. infusion as compared with daily s.c. injections of GH. Concentrations of the high density lipoprotein (HDL) cholesterol were significantly lower (p < 0.02) after the continuous GH pattern. Similarly, the HDL fraction Apo A-1 tended to be lower with constant GH delivery (p = 0. 052). Serum levels of total cholesterol, triglyceride and Apo B were similar on the two occasions. CONCLUSION: Short-term GH administration to GH-deprived GHD patients increased serum Lp(a), but only significantly with continuous delivery. During more prolonged GH exposure, constant s.c. infusion of GH resulted in slightly raised Lp(a) levels and reduced HDL and Apo A1 levels as compared with intermittently administered GH. The findings are consistent with the more effective induction of serum IGF-I levels after continuous patterns of GH delivery previously reported in GHD patients. Longer-term data are needed before conclusions with respect to the impact of the pattern of GH administration on, e.g., the risk of developing coronary heart disease can be drawn. PMID- 9873198 TI - Systemic administration of epidermal growth factor reduces fat mass in rats: effects on the hormone-sensitive-lipase, lipoprotein lipase and leptin. AB - We examined whether the reduction in fat mass induced by EGF treatment in mature animals was via activation of hormone-sensitive lipase (HSL) and thereby the induction of lipolysis, or through inhibition of lipoprotein lipase activity thus reducing fat uptake in adipose tissue. Sixteen male rats were treated with placebo or EGF 150 microg/kg/day for 7 days via mini-osmotic pumps. The results demonstrate that systemic EGF treatment reduces the amount of adipose tissue, most likely due to increased lipolysis as HSL activity as well as HSL mRNA were increased. The circulating levels of free fatty acids were slightly increased and leptin levels reflected the decrease in adipose tissue mass. PMID- 9873199 TI - The in vitro effect of leptin on basal and growth hormone-releasing hormone stimulated growth hormone secretion from the ovine pituitary gland. AB - We have studied the direct effect of leptin on the secretion of GH from the anterior pituitary gland of the sheep. Using primary cultures of ovine pituitary cells, leptin (10(-9)-10(-7) M) treatment for 30 min did not affect basal or GHRH (10(-7) M)-stimulated GH secretion. Following treatment for 24 h, a dose of 10( 7) M leptin stimulated basal GH secretion. In contrast, doses of 10(-7) and 10( 8) M leptin inhibited GHRH-stimulated GH secretion after 24 h of treatment. These results suggest that leptin can have long-term effects on the somatotropes, but no acute effect. Furthermore, leptin appears to have opposite effects on basal and GHRH-stimulated GH secretion. PMID- 9873200 TI - Role of phosphodiesterases in the regulation of gonadotropin- releasing hormone secretion in GT1 cells. AB - Increases in the level of cAMP stimulate the secretion of GnRH from GT1 GnRH neuronal cells. We hypothesized that cyclic nucleotide phosphodiesterases (PDEs), the enzymes that hydrolyze cAMP, may constitute a negative feedback signaling mechanism for GnRH regulation by decreasing the level of cAMP. GT1 cells were shown to express three PDEs by RT-PCR analysis: the cAMP-specific PDE4B and PDE4D and the calmodulin-dependent PDE1B. A splice variant of PDE4D, PDE4D3, which is activated when phosphorylated by cAMP-dependent protein kinase (PKA), was identified in GT1 cells by Western analysis. Consistent with PDEs negatively regulating GnRH secretion, treatment with the nonselective PDE inhibitor, IBMX, stimulated GnRH secretion 137% in 30-min static cultures. Furthermore, treatment with the PDE4-specific inhibitors Rolipram and RS-25344 increased GnRH secretion 48 and 125%, while treatment with the PDE1-specific inhibitor 8-MeoM-IBMX only caused a modest increase of 28%. In perifusion studies a rapid multi-fold stimulation of GnRH secretion was observed following treatment with IBMX, Rolipram or RS-25344. In conclusion, the level of PDE activity appears to be an important negative feedback signal for GnRH secretion. We hypothesize that activation of PDE4D3 by PKA may constitute a negative feedback signaling pathway which participates in the regulation of cAMP levels. PMID- 9873201 TI - Hypothalamic-pituitary-gonadal axis in the mutant weaver mouse. AB - The weaver (wv) mutant mouse manifests severe locomotor defects, a deficiency in granule cells of the cerebellum, and cellular deficits in the midbrain dopaminergic system. The wv phenotype is associated with a missense mutation in the pore region of the G-protein-gated inwardly rectifying potassium channel, GIRK2. The homozygous male wv mouse is essentially infertile due to an inadequate level of sperm production. Females are fertile although they also manifest the neurological phenotype. Homozygotes of both sexes have reduced body weight. We have evaluated the hypothalamic-pituitary-gonadal axis in heterozygote and homozygote male and female wv mutants in comparison with wild-type controls. Testicular weight was significantly reduced in the homozygous males, due to degenerative changes of seminiferous epithelium. Serum and pituitary content of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin were normal in all groups, and the normal sex differences were noted (FSH and LH higher in males, prolactin higher in females). Pituitary growth hormone (GH) concentration was normal, with control and mutant males showing higher GH than females. Serum testosterone levels were normal in the mutants, as was testicular testosterone. Testicular alpha-inhibin content was mildly reduced, but high in proportion to testicular weight. The defect in spermatogenesis appeared predominantly in the postmeiotic stages. In situ hybridization was consistent with expression of some GIRK2 mRNA isoforms in seminiferous epithelium. There were no significant differences between genotypes in the levels of dopamine, dihydroxyphenylacetic acid, serotonin and 5-hydroxyindoleacetic acid in the mediobasal and preoptic hypothalamic regions. Homovanillic acid levels in these two areas were, however, reduced in wv homozygotes compared to wild-type animals. In the light of normal pituitary hormone levels, normal hypothalamic monoamine concentrations and normal sex differences in gonadotropins, we conclude that the infertility in the male homozygote wv mouse lies within the tubule and is probably a primary defect in the germ cells. The hormonal data suggest that Leydig cell function, and at least some aspects of Sertoli cell function, are normal in the mutant mice. PMID- 9873202 TI - Delta opiate receptors account for the castration-induced unmasking of gonadotropin-releasing hormone binding sites in the rat pituitary. AB - Under control incubation conditions, gonadotropin-releasing hormone (GnRH) binds only a fraction of its receptors in rat-cultivated pituitary cells. Unmasking of the remaining receptors, which have been termed 'cryptic', requires drug- or peptide-induced protein kinase activation. Spontaneous masking however is not observed on pituitary cells sampled from castrated male rats, suggesting the presence of an intrinsic unmasking factor. Many endogenous factors could theoretically account for the effect. Here we attempted to identify the factor involved by taking advantage of their differential dependency upon second messengers and transduction cascades. Spontaneous unmasking of GnRH binding was found reversed by pertussis toxin (PTX), an inhibitor of alphai and alphao subunits of heterotrimeric G proteins, and by U73122, a phospholipase C (PLC) inhibitor. In contrast, desensitization of protein kinase C (PKC) or inhibition of tyrosine kinase by herbimycin were ineffective. Among endogenous pituitary factors able to unmask GnRH receptors in pituitary cells from normal male rats, as EGF, NPY or opiate peptides, only the latter were found to correspond to this transduction profile. In an attempt to characterize the pharmacology of opiate effects, naloxone (10 microM), a poorly selective opiate antagonist, restored masking of GnRH binding in cells from castrates. Only the delta antagonist naltrindole (1 microM) was able to mimick the action of naloxone. Conversely, when tested on cells from intact animals, morphine (10 microM), as well as dslet (1 microM) and met-ENK (10 nM), preferential delta agonists, but not dago and beta-endorphin or U50488 H and dynorphin, respectively micro and kappa agonists, were able to suppress masking. Among opioid peptides endogenous to the pituitary, only met-ENK was able to unmask cryptic receptors, an effect antagonized by naltrindole. We conclude that an opiate delta receptor subtype is endogenously activated in the pituitary of castrated male rats to prevent masking of GnRH binding. PMID- 9873203 TI - Ontogeny of the diurnal rhythm of tuberoinfundibular dopaminergic neuronal activity in peripubertal female rats: possible involvement of cholinergic and opioidergic systems. AB - The ontogeny of the diurnal change of tuberoinfundibular dopaminergic (TIDA) neuronal activity in female rats and its control mechanism were the foci of this study. Intact pre (28- and 35-day-old)-, peri (39- and 42-day-old)- and post (49- and 56-day-old)-pubertal female, as well as pre (28-day-old)- and post (56-day old)-pubertal male Sprague-Dawley rats were used. Basal TIDA neuronal activity, determined by measuring 3, 4-dihydroxyphenylalanine or 3,4-dihydroxyphenylacetic acid in the median eminence, increased 3- to 4-fold in the female and less than 2 fold in the male rats from day 28 to 49; so did serum prolactin levels. The diurnal rhythm of TIDA neuronal activity was not evident until day 42 in the female, but not male, rats. Neither nigrostriatal, nor mesolimbic DA neurons exhibited any significant change between pre- and postpubertal stages. In postpubertal (56-day-old) female rats, injection of nicotine (10 microg/kg) in the morning, but not in the afternoon, inhibited TIDA neuronal activity while similar injections of mecamylamine (1 mg/kg) or naloxone (2.5 mg/kg) stimulated these neurons in the afternoon, but not in the morning. Serum prolactin levels changed accordingly. In contrast, none of the injections had any effect in prepubertal (28-day-old) female rats. These results indicate that the genesis of the diurnal rhythm of TIDA neuronal activity in female rats occurs during the peripubertal stage, and endogenous cholinergic and opioidergic neurons may play a significant role. PMID- 9873204 TI - Expression of hypothalamic peptides in mice lacking neuronal nitric oxide synthase: reduced beta-END immunoreactivity in the arcuate nucleus. AB - The gas nitric oxide (NO) is an important messenger in brain signaling. Along with many other functions, NO is thought to influence the expression and/or release of various hypothalamic hormones (corticotropin-releasing hormone (CRH), gonadotropin-releasing hormone (GnRH) and vasopressin). To learn more about the role of NO in neuroendocrine mechanisms, we studied in mutant mice lacking neuronal isoform of NO synthase (nNOS) the cellular expression of CRH, neurophysin (the carrier protein of vasopressin/oxytocin) and pro opiomelanocortin (POMC), as well as of the POMC-derived peptides beta-endorphin (beta-END), alpha-melanocyte-stimulating hormone (alpha-MSH) and corticotropin (ACTH) by use of immunohistochemistry and in situ hybridization. Additionally, the remaining NO-generating capacities of the nNOS minus mice were investigated by NADPH-diaphorase histochemistry and citrulline immunohistochemistry as well as by immunohistochemical localization and Western blot analysis of endothelial NOS (eNOS) and nNOS isoforms. Amongst all hypothalamic peptides under investigation, only beta-END was found to be altered in mutant mice. A morphometric analysis of beta-END producing neurons of the arcuate nucleus revealed that significantly less cells were immunoreactive in mutant mice, whereas the expression of the precursor POMC as well as of other POMC-derived peptides was found to be unchanged. In addition to that, fewer beta-END-immunoreactive fibers were found in the paraventricular nucleus of nNOS minus mice in comparison to wild-type animals. Hence, the reduction of hypothalamic beta-END is probably a posttranslational event that might reflect a disturbed endorphinergic innervation of those hypothalamic neurons which normally express nNOS. PMID- 9873205 TI - Ultrastructural immunolocalization of leptin receptor in mouse brain. AB - Antibodies directed to amino acids 877-894 (M-18) and 32-51 (K-20) were used to localize leptin receptor by immunocytochemistry in mouse brain. Both antibodies stained several hypothalamic nuclei (paraventricular nucleus, supraoptic nucleus, supraoptic retrochiasmatic nucleus, suprachiasmatic nucleus, preoptic area, ventromedial nucleus, dorsomedial nucleus, lateral hypothalamus, arcuate nucleus, ventral and dorsal premammillary nuclei), the thalamic and amygdaloid nuclei, neurons of the neocortex and archicortex and the epithelial cells of the choroid plexus. While M-18 staining was concentrated in the Golgi area, with K-20 it was dispersed in the cytoplasm. Glial cells were stained only by K-20. These results suggest that the trans-membrane forms of the receptor are concentrated at the membrane level of the Golgi complex of neurons and in epithelial cells of the choroid plexus while the soluble form is dispersed in their cytoplasm. Glial cells express only the soluble form. PMID- 9873206 TI - Transcription factor STAT3 in leptin target neurons of the rat hypothalamus. AB - Leptin is an adipose tissue-derived hormone that regulates body weight via interactions with hypothalamic neuronal circuitries expressing specific leptin receptors (Ob-R). The Ob-Rs act via the JAK-STAT (Janus kinase-signal transducers and activators of transcription) pathway of signal transduction. Recent evidence suggests that primarily the transcription factor STAT3 mediates leptin's action in the hypothalamus. We have investigated the presence and cellular localization of STAT3 protein in the rat hypothalamus by means of indirect immunofluorescence histochemistry using a rabbit polyclonal STAT3 antiserum. The antiserum identified a 92-kDa protein using Western blotting on rat hypothalamic homogenates, corresponding to the expected size of STAT3. STAT3 immunoreactivity was demonstrated in Ob-R-containing neurons of the paraventricular nucleus (parvocellular part), periventricular nucleus, arcuate nucleus and in the lateral hypothalamic area. Direct double-labeling showed presence of STAT3 immunoreactivity in neuropeptide Y (NPY)-containing neurons of the ventromedial part of the arcuate nucleus and in proopiomelanocortin (POMC)-containing neurons of the ventrolateral part of the arcuate nucleus. The results provide an anatomical basis for a leptin action mediated by STAT3 in Ob-R-containing NPY and POMC neurons of the arcuate nucleus, as well as by Ob-R-containing neurons of the parvocellular paraventricular nucleus and lateral hypothalamic area. PMID- 9873207 TI - Thyroid hormone-dependent regulation of galanin synthesis in neurons and glial cells after colchicine administration. AB - Galanin and galanin message-associated peptide (GMAP) synthesis is up-regulated in neurons and glial cells in the adult rat brain by different experimental manipulations, including intracerebroventricular colchicine. We have previously reported that the galanin expression is severely attenuated in some neurons in adult hypothyroidism. In order to further investigate the role of thyroid hormone for the in vivo regulation of galanin gene expression, we have studied the effect of intraventricular administration of colchicine on prepro-galanin (ppGAL) mRNA expression in the brain of normal and hypothyroid, adult male rats. While ppGAL mRNA levels were markedly elevated in a great number of glial cells in the white and gray matter in normal rats, this effect was almost completely abolished in hypothyroid rats. In contrast, colchicine-induced up-regulation of galanin/GMAP expression occurs in the hypothalamic paraventricular nucleus both of euthyroid and hypothyroid rats, although with a slightly different time course. PMID- 9873208 TI - Renal handling of calcium. AB - Ca absorption is mediated by both active and passive processes. Absorption in both proximal tubule and thick ascending limb is mainly coupled indirectly to Na absorption and is a passive process through the paracellular pathway. In the distal convoluted tubule, Ca absorption is regulated independently of Na absorption; this is the principal site of action of parathyroid hormone, calcitonin, and 1,25-(OH)2D3. Models for transport in proximal tubule, thick ascending limb, and distal convoluted tubule afford attractive explanations for the influence on Ca transport of effective arterial blood volume, parathyroid hormone, acid-base balance, genetic disturbances such as Bartter's and Gitelman's syndromes, and diuretic action. PMID- 9873209 TI - Cell-crystal interactions and kidney stone formation. AB - BACKGROUND: Renal tubular fluid in the distal nephron is supersaturated with calcium and oxalate ions that nucleate to form crystals of calcium oxalate monohydrate (COM), the most common crystal in renal stones. How these nascent crystals are retained in the nephron to form calculi in certain individuals is not known. METHODS: The results of experiments conducted in this and other laboratories that employ cell culture model systems to explore renal epithelial cell-urinary crystal interactions are described. RESULTS: COM crystals rapidly adhere to anionic sites on the surface of cultured renal epithelial cells, but this process can be inhibited, if specific urinary anions such as glycosaminoglycans, uropontin, nephrocalcin, or citrate are available to coat the crystalline surface. Therefore, competition for the crystal surface between soluble anions in tubular fluid and anions on the apical cell surface could determine whether or not a crystal binds to the cell. A similar paradigm describes adhesion of calcium phosphate (hydroxyapatite) crystals, also a common constituent of human stones. Once bound, COM and hydroxyapatite crystals are quickly internalized by renal cells; reorganization of the cytoskeleton, alterations in gene expression, and initiation of proliferation may then ensue. Each of these cellular events appears to be regulated by a different set of extracellular factors. Over several weeks in culture, renal cells (BSC-1 line) dissolve internalized crystals, although once a cell binds a crystal, additional crystals are more likely to bind, possibly forming a positive feedback loop that results in kidney stone formation. CONCLUSIONS: Increased knowledge about the cell-crystal interaction, including identification of molecules in tubular fluid and on the cell surface that modulate the process, and understanding its mechanism of action appear critical for explaining the pathogenesis of nephrolithiasis. PMID- 9873211 TI - Epidemiology of nephrolithiasis. AB - The main purpose of the study is to acquire useful information on nephrolithiasis in Italy, given the recent increase in its prevalence. The analysis was based on data from sample surveys of the Italian population carried out in 1983 and 1993 1994 as well as on data obtained from medical records of patients treated at the ASTIF in Fiuggi. An increase in the prevalence of the disease from 11.7 to 17.2 out of 1,000 inhabitants has been observed over the last 10 years; most patients are men and elderly, and the highest frequency is in southern and the lowest in northern Italy. The results of the study confirm the hereditary predisposition, since nephrolithiasis is very common among patients' parents (22.5%) and siblings (14.1%). The patients eat more meat than others and are often overweight or obese; the latter represent 18% of these patients and 11% of the general population. PMID- 9873210 TI - Relationship between urinary calcium and net acid excretion as determined by dietary protein and potassium: a review. AB - Increasing urinary net acid (titratable acid + NH4 - HCO-3) excretion is accompanied by an increased urinary Ca excretion because of reduced renal tubular reabsorption of filtered Ca. The relationships between urinary Ca excretion rates and urinary net acid excretion rates are reviewed for data: (1) among healthy adults eating constant diets when net acid excretion is increased by increasing dietary protein, administering NH4Cl, or withdrawal of dietary KHCO3 or reduced by administering KHCO3; (2) among healthy adults eating constant diets providing varying amounts of protein and potassium, and (3) among healthy adults and Ca stone formers with and without idiopathic hypercalciuria eating ad libitum. The results show that urinary Ca excretion varies directly with net acid excretion by 0.035 mmol/mEq. The urinary net acid excretion increases by 0.10-0.15 mEq/mmol urinary urea, and urinary Ca increases by about 0.04 mmol/g dietary protein, while the urinary net acid excretion decreases as the ratio of urinary K/urea, a reflection of the dietary K relative to dietary protein, increases. The relationships between net acid excretion and both urinary urea and K/urea are similar among Ca stone formers without and with idiopathic hypercalciuria, but those with idiopathic hypercalciuria exhibit increased rates of urinary Ca excretion at all levels of net acid excretion. PMID- 9873212 TI - Urine volume: stone risk factor and preventive measure. AB - BACKGROUND: A high fluid intake is the oldest existing treatment for kidney stones, and, up until a few decades ago, it was the only preventive measure at the physician's disposal for stone recurrences. METHODS: Using the data available in literature and partly unpublished personal research, we examine the role of urine volume as a stone risk factor, its impact on calcium crystallization mechanisms and its real importance as a means of prevention. RESULTS: To sum up, the most important findings are: (1) a low urine volume must be considered as a real risk factor, both as regards the onset of renal calculi and stone relapses; (2) an increase in urine volume induced by a high water intake produces favourable effects on the crystallization of calcium oxalate and does not reduce the activity of natural inhibitors; (3) a sufficiently high intake of water and probably other fluids such as coffee, tea, beer and wine has a preventive effect on nephrolithiasis and its recurrence, and (4) the role of fruit juice is still to be defined. CONCLUSIONS: A high intake of fluids, especially water, is still the most powerful and certainly the most economical means of prevention of nephrolithiasis, and it is often not used to advantage by stone formers. PMID- 9873213 TI - Idiopathic calcium nephrolithiasis. AB - Calcium nephrolithiasis (CaNL) accounts for more than 70% of all renal stones, and its prevalence has increased in the last decades. Under this definition are included patients passing stones, composed of calcium oxalates and/or calcium phosphates. Current views of the pathogenesis of CaNL are based on the role of metabolic abnormalities which concur to render urines more conducive to crystallization. Therefore, the diagnostic approach is aimed at detecting these abnormalities, and the medical treatment assumes that a decrease in the risk of lithogenesis will result in remission or improvement of recurrences. The workup of the patients with CaNL begins with the analysis of passed stones and X-ray, sonography or other imaging techniques. Eligible patients, that is, both recurrent active stone formers and single-stone formers with individual risk factors, are considered for a metabolic evaluation, by which a number of blood and urine parameters are measured and others calculated. These include estimates of urine state of saturation with calcium and uric acid salts, net gastrointestinal alkali absorption, renal threshold of phosphate and other renal clearances and net acid and total nitrogen excretions. Basically, this screening is informative on renal function, metabolic abnormalities and their pathophysiology, risk of stone formation and dietary habits. During treatment it gives information about patient compliance and adverse effects of therapy. The cost of a comprehensive screening in Piedmont is 192,000 ITL (100 Euro) and rises to 300,000 ITL (154 Euro) if hormones and hydroxyproline are measured. In individual patients second- and third-level studies are performed, in order to detect systemic diseases which account for about 20% of CaNL in our series. Cost to-benefit analysis has shown that the medical procedures for CaNL yield considerable saving in terms of difference between expenditure for drugs and testing and reduction of stone events. However, the current workup cannot be considered exhaustive, because misleading events may hamper the relation between laboratory findings and clinical outcome, and factors other than urine composition have appeared on the scenario of nephrolithiasis. These represent our challenge for the third millennium. PMID- 9873214 TI - Hyperuricemic nephropathies. AB - This review explores the relationship between uric acid or urate and the pathogenesis of renal impairment. The following points and conclusions are emphasized: (1) uric acid is an end product of purine degradation in humans and normally depends upon renal excretion for the majority of its elimination from the body; (2) massive urate overproduction - usually occurring acutely because of tumor lysis, rhabdomyolysis, or some other cause of rapid nucleic acid turnover or tissue destruction - tends to cause acute renal failure because of an increase of intratubular uric acid precipitation and obstruction; (3) chronic urate overproduction (with increased urate excretion) is more likely to be associated with stones or gout than with acute renal failure; (4) chronic asymptomatic hyperuricemia is unlikely to cause renal disease, gout, or stones, but is associated with cardiovascular impairment over the long term, and (5) asymptomatic hyperuricemia may serve as an indicator of renal vascular disease, or, to the extent that it may reflect insulin-induced acceleration of renal tubule urate reabsorption, hyperuricemia may serve as an indicator of insulin resistance. Therefore chronic asymptomatic hyperuricemia may predict the adverse cardiovascular consequences of insulin resistance. PMID- 9873216 TI - Medical prevention of renal stone disease. AB - Medical treatment designed to prevent stone formation is important in idiopathic calcium oxalate nephrolithiasis, because of the high rate of stone recurrence. Several randomized trials have established the values of conservative and drug treatments. A high fluid intake alone has been reported to inhibit the recurrence of stone formation in single stone formers. In patients with recurrent disease, a significant reduction in stone formation rate from pretreatment was found in the placebo group maintained on a conservative program, underscoring the importance of increased fluid intake and dietary modification. In patients with active recurrent stone disease, treatment with drugs along with a conservative program is necessary. Allopurinol, thiazide, potassium citrate and potassium-magnesium citrate have been shown to inhibit stone formation compared with placebo. It has not been clearly established that a selective treatment is more effective than a more randomly chosen drug treatment. Another advantage of medical approach is its ability to correct nonrenal complications of stone disease, such as bone loss that sometimes accompanies stone disease. PMID- 9873215 TI - Struvite stones. AB - Struvite stones constitute only about 2-3% of the stones reaching the laboratory for analysis, but the clinical problems they create including sepsis and even renal demise are greater than with any other stone type. This article reviews the evidence that bacterial urease, usually from a Proteus species, is responsible for the chemical changes in urine which result in struvite formation. Available urease inhibitors and other forms of medical management of patients with these stones are discussed. A patient with struvite stones should be assumed to have a progressive disease which cannot be ignored. Even after seemingly successful elimination of stones with lithotripsy and/or percutaneous nephrolithotomy, careful medical follow-up is critical. The medical profession is probably underutilizing postprocedure hemiacidrin irrigation because of shortsighted financial considerations. Primary-care physicians need to be educated in the importance of aggressive management of Proteus and other urea-splitting infections. PMID- 9873217 TI - Effects of water hardness on urinary risk factors for kidney stones in patients with idiopathic nephrolithiasis. AB - Both amount and timing of dietary calcium intake influence the recurrence of renal calcium stones. We have evaluated whether the hardness of extra meal drinking water modifies the risk for calcium stones. The urinary levels of calcium, oxalate and citrate, i.e., the main urinary risk factors for calcium stones, were measured in 18 patients with idiopathic nephrolithiasis, maintained at fixed dietary intake of calcium (800 mg/day), after drinking for 1 week 2 liters per day, between meals, of tap water and at the end of 1 week of the same amount of bottled hard (Ca2+ 255 mg/l) or soft (Ca2+ 22 mg/l, Fiuggi water) water, in a double-blind randomized, crossover fashion. As compared with both tap and soft water, hard water was associated with a significant 50% increase of the urinary calcium concentration in the absence of changes of oxalate excretion; the calcium-citrate index revealed a significant threefold increase during ingestion of hard water as compared with respect to soft water (Fiuggi water), making the latter preferable even when compared with tap water. This study suggests that, in the preventive approach to calcium nephrolithiasis, the extra meal intake of soft water is preferable to hard water, since it is associated with a lower risk for recurrence of calcium stones. PMID- 9873218 TI - Treatment of renal stones by extracorporeal shock wave lithotripsy. AB - Based on an extensive review of the literature and our own clinical experience, this article attempts to present clear guidelines for the management of various kidney stones that will be acceptable to clinical urologists and their patients. Regarding our own patients, we compared different studies and discussed the results concerning the anatomical kidney situation, stone size, stone localization and observation time. Stone-free rates of patients with calyceal diverticula calculi range from 4 to 58%, with an increase after longer follow-up periods. According to the importance of residual fragments following extracorporeal shock wave lithotripsy (ESWL), we have to distinguish between clinical insignificant residual fragments and clinical significant residual fragments. 24 months following ESWL, stone passage occurs as a continuous process, and if there are no clinical symptoms, any endoscopic procedure should be considered as over-treatment. Newer ESWL technology has increased the percentage of clinically insignificant residual fragments. We consider percutaneous nephrolithotripsy in most of the patients with renal calculi smaller than 30 mm in diameter only as second-line therapy. PMID- 9873219 TI - Ecosystem of the Anticolana Valley. AB - The results of an integrated environmental study on the geological, hydrogeological and ecological features of the area of formation of the low mineral content water of Fiuggi are briefly presented. The research programme, that was started in 1996 and is now in its final phase, was aimed at the following: (a) identifying the geological-structural limits of the hydrogeological basin; (b) building a model of the deep water circulation that should include an analysis of the rainfall and of the water balance, and (c) determining the vulnerable characteristics of the aquifer in relation to the human impact and setting up a monitoring system to control the different environmental parameters. PMID- 9873220 TI - Chemical analysis of water of the Anticolana Valley: isolation of humic compounds. AB - The purpose of this study was to identify and characterize humic compounds in Anticolana Valley (Fiuggi) water. The capacity of this class of compounds to mobilize metals from solid phases could have an important role in calculi solubilization. Humic compounds were isolated, purified and characterised by FTIR, 1H-NMR and 13C-NMR spectroscopy, thermogravimetry and elemental analysis. Only fulvic acids were found. They are mainly composed of aliphatic chains, made of six -CH2O- groups and contain a number of carboxylic groups, responsible for their metal complexing capacity. PMID- 9873221 TI - Solvent effect in vitro of Anticolana Valley water on renal stones: analytical instrumental study. AB - The presence of humic and fulvic acids in the Anticolana Valley (Fiuggi) water has been established. On the basis of this evidence we investigated the capacity of Anticolana Valley (Fiuggi) water to dissolve renal calculi in vitro. Crystals of calcium oxalate monohydrate to simulate a kidney stone were prepared. Human renal stones of calcium oxalate monohydrate were obtained by courtesy of the Division of Urology of 'La Sapienza' University (Rome), the Division of Urology of the University of Havana (Cuba) and the ASTIF of Fiuggi. The study was performed using the Anticolana Valley (Fiuggi) water, distilled water and tap water (ACEA, Rome), in a specially designed Perspex apparatus. Each calculus was subjected to a water flow of 2 liters/24 h. The capacity of the Anticolana Valley (Fiuggi) water to dissolve human and synthetic calculi was found to be much higher than that of distilled water which in turn was significantly more effective than tap (ACEA) water. PMID- 9873222 TI - T cells mediate treatment of six-day-old cytokine-gene-transfected mouse mammary tumor. AB - We have previously shown that immunologically different mouse mammary cancer cell lines induce antitumor responses after IL2 or IL4 gene transfection. We now report the ability of cytokine-transfected tumors to induce eradication of established wild-type tumor. Animals with 6-day-old tumor treated with IL2 transfected cells also had significantly smaller tumors 2.8 and 1.7 cm2 (EMT6 and 410.4). Findings were similar for IL4-transfected cells. Tumor infiltrating lymphocytes or cells from draining lymph nodes demonstrated tumor-specific in vitro cytotoxicity. Immunohistochemical studies revealed T cell infiltrates in transfected tumors. PMID- 9873223 TI - Role of interleukin-8 in tumor growth and metastasis of human melanoma. AB - Expression of interleukin-8 (IL-8) by human melanoma cells correlates with their metastatic potential in vivo. Moreover, UVB irradiation of primary cutaneous melanoma induces IL-8 mRNA and protein production and increases both tumor growth and metastasis in nude mice. Although IL-8 has been shown to be an angiogenic factor, the biological consequences of increased IL-8 production by melanoma cells and the role of IL-8 in the metastatic process remains unclear. The purpose of this review is to determine the role of IL-8 in tumor growth and metastasis of human melanoma. Transfection of nonmetastatic and IL-8-negative melanoma cells with the IL-8 gene rendered them highly tumorigenic and increased their metastatic potential in nude mice. The IL-8-transfected cells displayed upregulation of MMP-2 expression and activity and increased invasiveness through Matrigel-coated filters. Activation of MMP-2 by IL-8 can enhance the invasion of host stroma by the tumor cells and increase angiogenesis and, hence, metastasis. In addition to UVB, IL-8 can also be upregulated by hypoxia conditions, suggesting that the environment plays a major role in regulating IL-8 expression and metastasis. The studies summarized in this review suggest that in melanoma, IL-8 may serve as the angiogenic factor distinguishing benign from malignant cells. PMID- 9873224 TI - Enhancement of atherosclerosis in beta-2-glycoprotein I-immunized apolipoprotein E-deficient mice. AB - We have previously shown that low density lipoprotein receptor-deficient (LDL-RD) mice immunized with beta2-glycoprotein I (beta2GPI; a target of autoimmune anticardiolipin antibodies) developed enhanced early atherosclerosis, when fed a normal chow diet. The current study was undertaken to evaluate the effect of immunization with beta2GPI and the addition of a high fat diet on the progression of atherosclerosis in the apolipoprotein E (ApoE)-deficient mouse. Six-week-old female ApoE-deficient mice (n = 10) were immunized subcutaneously with either human beta2GPI or with ovalbumin, both emulsified in complete Freund's adjuvant and fed a high fat diet for 6 weeks. The beta2GPI-immunized mice were found to develop accelerated atherosclerosis when compared with their ovalbumin-immunized littermates (aortic lesion area of 137,500 +/- 13,801 vs. 72,444 +/- 14,465 microm2, respectively; p = 0.0067). The beta2GPI-immunized mice developed high titers of anti-beta2GPI antibodies, 10 days after the procedure, which were sustained until the sacrifice. LDL extracted from both study groups displayed similar susceptibility to ex vivo oxidation. These results confirm our previous study in which we found increased atherosclerosis in beta2GPI-immunized LDL-RD mice fed a chow diet. In the current study we show that the proatherogenic effect of beta2GPI immunization is maintained despite high cholesterol levels and is not associated with increased susceptibility of LDL to ex vivo oxidation. PMID- 9873225 TI - Stereology and immunohistochemistry of the myocardium in experimental hypertension: long-term and low-dosage administration of inhibitor of the nitric oxide synthesis. AB - Morphological changes in the myocardium after left ventricular hypertrophy, due to chronic experimental hypertension, require an understanding of the quantitative relationship between myocyte and nonmyocyte compartments forming the structural framework of the myocardium. Hypertension was induced by long-term low dosage inhibition of nitric oxide synthesis by NG-nitro-L-arginine methyl ester (L-NAME) in rats. L-NAME (12 mg/kg) was given to animals in water ad libitum during 15 weeks. After this period, systolic blood pressure increased almost 50% as compared with that in the control group. Morphological changes in control and L-NAME animals were investigated with stereology and immunohistochemistry. Comparing control and L-NAME animals, the surface density of myocytes decreased 73.7% while the mean cross-sectional area increased 97.6% in L-NAME rats. The volume density of myocytes decreased 45.9% and the volume density of the interstitium increased 71.7% in L-NAME rats. No stereological difference was found in blood vessels comparing the two groups. Remodeling of the cardiac interstitium occurred with increased deposition of both fibronectin and type III collagen. Fibronectin was seen in both early and latter responses to infarction while type III collagen was seen mainly in areas of incomplete healing among myocytes and around intramyocardial branches of the coronary arteries. The long term low-dosage administration of an inhibitor of the NO synthase such as L-NAME causes myocyte hypertrophy and early interstitial and perivascular fibrosis without important quantitative changes in microcirculation. PMID- 9873226 TI - Long-term telomere dynamics: modest increase of cell turnover in HIV-infected individuals followed for up to 14 years. AB - To quantify the long-term dynamics of telomere lengths and the effect of HIV infection on lymphocyte turnover rates, we measured in a blinded study longitudinal samples from 6 individuals using a highly accurate method based on two-dimensional calibration of DNA sizes. For two uninfected controls followed 8 and 10 years the average telomeric terminal restriction fragment (TRF) shortening rate in peripheral blood mononuclear cells (PBMCs) was 50 and 60 bp/year, respectively, in agreement with previous measurements of cross-sectional samples. The TRF lengths of PBMCs from two slow progressors followed for 14 years declined by a rate of 120 +/-10 bp/year, i.e. 2-fold higher than the rate of TRF shortening for uninfected individuals. The rate of TRF shortening was higher in CD8 (140 +/-10 bp/year) than in CD4 (100 +/-10 bp/year) cells. The CD8 cell TRFs of the two fast progressors shortened faster (240 +/-10 bp/year) and the rate of CD4 cell TRF shortening in one of the fast progressors was 160 bp/year. These data suggest that HIV infection causes only a modest increase in the lymphocyte turnover which we speculate could be due to chronic activation of the immune system, and may not result in the exhaustion of its regenerative capacity and immunopathogenesis. PMID- 9873227 TI - Cellular immune response to retinal S-antigen and interphotoreceptor retinoid binding protein fragments in Eales' disease patients. AB - The role of retinal antigens in Eales' disease was studied in 24 patients and an equal number of healthy controls. Lymphocyte proliferative responses were tested in vitro against native S-antigen, its uveitopathogenic peptides (peptide M and peptide G), yeast histone H3 peptide and uveitopathogenic fragment of interphotoreceptor retinoid-binding protein (IRBP; R16) to establish their role in the pathogenesis of Eales' disease. Out of 24 Eales' disease patients, 6 showed significant proliferative response against S-antigen, its uveitogenic fragments or IRBP. None among the controls showed any response to any retinal antigen used in this study. There was no statistically significant difference in the response to purified protein derivative between patients and controls. These results suggest that retinal antigens may play a role in the etiopathogenesis of Eales' disease. An extraneous agent that could result in exposure of normally sequestered uveitopathogenic antigens of the immune system, leading to an exuberant immune response in the eye may initiate the disease. PMID- 9873228 TI - Changes in keratin and filaggrin expression in the skin of chronic proliferative dermatitis (cpdm) mutant mice. AB - The expression of keratins and filaggrin by keratinocytes is a highly regulated process and depends on their state of differentiation and proliferation. As such, these proteins can be used as markers to determine if keratinocyte differentiation is normal. Mutant cpdm/cpdm mice develop a chronic skin disease characterized by epidermal hyperplasia and inflammation. Immunohistochemical staining for the basal keratins K5 and K14 revealed expression in the basal and suprabasal cell layers. The expression of K1 and K10 was reduced and limited to the outer layers of the stratum spinosum. Keratin 6 was expressed in the suprabasal layers of affected skin, and throughout all layers in severely affected skin. Filaggrin was present in the stratum granulosum which had variable thickness. These results indicate that the differentiation of keratinocytes in cpdm/cpdm mice was normal. The altered distribution and expression of keratins in comparison with the skin of control mice was the result of hyperproliferation. PMID- 9873229 TI - Expression of substance P receptors in normal and psoriatic skin. AB - Recently, substance P receptors (SPR) have been detected in neonatal foreskin. Our purpose was to determine the expression of SPR in other localizations than neonatal foreskin. As SP has been implicated in the pathogenesis of inflammatory cutaneous lesions, we wondered whether SPR localization was modified in psoriatic lesions. In normal skin, SP binding sites were detected using biotinylated SP and abrogated by a specific NK1 antagonist (spantide) on blood vessels, sweat glands and hair follicles. In the normal epidermis, SPR were usually observed on granular layers but may also be observed on other cell layers. The SP binding processed on cultured keratinocytes demonstrated that SPR were expressed in the epidermis, except basal cell layers, confirming that keratinocytes constitutively express SPR. In skin lesions of psoriatic patients, SP binding sites were expressed on the uppermost keratinocytes which are not granular cells, and seem to be overexpressed. Our results raise the question of the role of SPR on psoriatic keratinocytes. PMID- 9873230 TI - Conventional and molecular cytogenetic identification of a variant klinefelter syndrome patient with a deleted X chromosome. AB - We report on the case of a 34-year-old patient with the Klinefelter syndrome and an unusual cytogenetic finding of a deletion involving the short arm of the X chromosome. This was confirmed with fluorescent in situ hybridization (FISH) using an X chromosome-specific whole chromosome painting probe. The patient presented with infertility. The only abnormal physical findings were atrophic testes with azoospermia and elevated levels of follicle-stimulating hormone and luteinizing hormone. This case represents a relatively mild manifestation of the Klinefelter syndrome. Previous reported cases were often associated with more severe phenotypes such as variable degrees of mental retardation and facial dysmorphism, hypothesized as due to the failure of X inactivation. The X inactivation center, located on Xq13, is presumably intact in our patient, who had a deletion involving only the short arm. The mild phenotype observed in our patient was found to be consistent with the conventional and molecular cytogenetic findings. PMID- 9873231 TI - Acute and chronic citalopram treatment differently modulates rat exploratory behavior in the exploration box test: no evidence for increased anxiety or changes in the [3H]raclopride binding. AB - The effect of acute and chronic desipramine (10 mg/kg) and citalopram (10 mg/kg) treatment on rat exploratory behavior in the recently developed exploration box test was studied on 5 consecutive days. Acute desipramine but not citalopram treatment decreased the time spent exploring, the number of line crossings, rears, investigative approaches, entries into the large arena, and sum of exploratory events. After 3 weeks of pretreatment, both desipramine and citalopram attenuated rat exploratory behavior, whereas the number of entries into the large arena was unchanged. In the open field test, acute desipramine or citalopram treatment (5, 10, 15 mg/kg) attenuated rat exploratory behavior in a dose-dependent manner. In the subsequent rota-rod test, neither desipramine nor citalopram treatment (0-20 mg/kg) impaired motor performance capacity. In an additional experiment, [3H]raclopride binding was unchanged after single as well as 3 weeks of desipramine or citalopram treatment in the rat brain neostriatum. Our experiments demonstrate that acute citalopram treatment in the open field test and chronic citalopram treatment in the exploration box test attenuate rat exploratory behavior, but these effects may not be implicated with enhanced anxiety or changed dopamine D2 receptor characteristics. PMID- 9873232 TI - Prostaglandin E2 in human placenta: its vascular effects and activation of prostaglandin E2 formation by nicotine and cotinine. AB - Tobacco smoking by pregnant women increases the frequency of spontaneous abortions and preterm births. Human labor is associated with enhanced intrauterine phospholipid metabolism and production of prostaglandin E2 (PGE2) which induces labor, initiates uterine contractions and maintains the homeostasis of placental blood flow. Therefore, we studied: (a) the influence of nicotine and cotinine on the effects of PGE2 on placental vasculature in perfused human placental cotyledon, and (b) the activation of placental phospholipase A2 (PLA2) by nicotine and cotinine using 1-palmitoyl-2-[1-14C]arachidonyl phosphatidylethanolamine (PE, 2.2 nmol) as substrate. These studies revealed that: (1) increasing concentrations of PGE2 (10- 150 ng/ml) increased umbilical perfusion pressure by 170 +/- 10% (n = 6) of control (100%). Cotinine (2 microg/ml) enhanced this effect at all concentrations of PGE2. Nicotine (2 microg/ml) prevented the effect of PGE2; (2) both cotinine (EC50 470-500 fmol/l) and nicotine (EC50 18-32 pmol/l) activated PLA2 in human placental tissues. These observations indicated that cotinine was more potent than in nicotine activating PLA2 and potentiating the vasoconstrictive effects of PGE2 on fetal placental circulation. Nicotine activates nicotinic receptors and releases placental acetylcholine, a vasodilator of placental arteries. Acetylcholine stimulates muscarinic receptors of endothelial cells resulting in the release of endothelium derived relaxing factor (EDRF), and possibly nitric oxide. Therefore, nicotine prevents or abolishes the vasoconstrictive effects of PGE2 through the release of EDRF. Cotinine is inactive at nicotinic and muscarinic receptors. Therefore, accumulation of cotinine, the major metabolite of nicotine, in fetal circulation may contribute to production of PGE2 and induction of preterm labor and spontaneous abortions. PMID- 9873233 TI - Subacute cocaine treatment changes expression of mouse liver cytochrome P450 isoforms. AB - Acute administration of single high doses of cocaine (50 or 60 mg/kg) produces liver injury in mice that have been pretreated with inducers of mixed function oxidases. Multiple low doses of cocaine (10-30 mg/kg) will produce hepatotoxicity without prior induction. To establish whether cocaine can induce its own activation, mice were given three daily injections of cocaine. Total cytochrome P450 content of the liver did not change. After 3 days the amount of cytochrome P450 2B10, as measured by pentoxy resorufin-O-dealkylase activity and immunoblotting, increased 3-fold. Cytochrome P450 2A5-catalyzed coumarin 7 hydroxylase activity and immunoreactive protein increased by about 50%. Enzyme activities and Western blotting of isoforms 1A, 2E, and 3A showed no change during this time. Chronic cocaine increased N-hydroxylation of norcocaine. Immunoinhibition studies showed that cytochrome P450 2A5 was the major isoform responsible for norcocaine N-hydroxylation. These results demonstrate that chronic cocaine can induce its own metabolism. Similar increases were also observed in mice not susceptible to liver injury from chronic cocaine. PMID- 9873234 TI - Effect of Am-80, a synthetic derivative of retinoid, on experimental arthritis in mice. AB - Am-80 is a newly snythesized retinoid with the structure of one aromatic amide among retinobenzoic acids. It exhibits specific biological activities of retinoic acid such as the activation of cellular differentiation and proliferation. We investigated the effect of Am-80 on collagen-induced arthritis (CIA) in mice and the immunopharmacological action on the production of several cytokines in the in vitro and in vivo models. Am-80, at doses of 0.3, 1 and 3 mg/kg, significantly inhibited the severity and development of the arthritis index, progression of foot pad swelling, bone damage and histopathological alterations. Am-80 also inhibited the production of anti-type II collagen (CII) IgG antibody, but did not affect the delayed-type hypersensitivity (DTH) response in arthritic mice. To determine the inhibitory mechanism of Am-80, we studied the effect of Am-80 on the production of cytokines. Am-80 did not affect the production of IFN-gamma by Th1 cells (1E10.H2 cells) and IL-4 by Th2 cells (D10.G4.1 cells), respectively. Am-80 selectively inhibited bacterial lipopolysaccharide (LPS)-induced IL-6, but not TNF-alpha and IL-1beta, production in mice. Moreover Am-80 inhibited IL-1beta induced IL-6 production and IL-6 mRNA expression in human osteoblast-like cells (MG-63). The inhibition of IL-6 production by Am-80 was due to downregulation of the pretranscription or the transcription of IL-6 in MG 63 cells. These findings suggest that the inhibitory effect of Am-80 on CIA is partially by modulating the production of the proinflammatory cytokine, IL-6. PMID- 9873235 TI - Lipowski's legacy: the psychosomatic spirit. PMID- 9873236 TI - Raising questions about antidepressants. AB - Antidepressant medication has apparently become the most popular treatment for depression in the USA. Several beliefs about the efficacy of antidepressant medications prevail among mental health professionals and the public. This paper explores relevant research data and raises questions about these beliefs. Many of the common beliefs about these medications are not adequately supported by scientific data. The following issues are raised: (1) industry-funded research studies which result in negative findings sometimes do not get published; (2) placebo washout procedures may bias results in some studies; (3) there are serious questions about the integrity of the double-blind procedure; (4) the 'true' antidepressant drug effect in adults appears to be relatively small; (5) there is minimal evidence of antidepressant efficacy in children; (6) side effects are fairly common even with the newer antidepressants; (7) combining medications raises the risk for more serious complications; (8) all antidepressants can cause withdrawal symptoms; (9) genetic influences on unipolar depression appear to be weaker than environmental influences; (10) biochemical theories of depression are as yet unproven; (11) biological markers specific for depression have been elusive; (12) dosage and plasma levels of antidepressants have been minimally related to treatment outcome; (13) preliminary evidence suggests that patients who improve with cognitive-behavioral psychotherapy show similar biological changes as those who respond to medication, and (14) the evidence suggests that psychological interventions are at least as effective as pharmacotherapy in treating depression, even if severe, especially when patient rated measures are used and long-term follow-up is considered. PMID- 9873237 TI - Cognitive-behavioral therapy for idiopathic infertile couples. AB - BACKGROUND: The purpose of this pilot study was to evaluate the impact of a 6 month cognitive-behavioral therapy for infertile couples. METHODS: Seventeen idiopathic infertile couples participated in a therapy program comprised of modules to behaviorally optimize the chance of conception, improve sexual functioning and satisfaction, reduce thoughts of helplessness and, if necessary, improve marital communication skills. Pre- to posttreatment changes in the therapy group were compared to changes in two control groups. RESULTS: The therapy group showed an improvement in sperm concentration, a reduction in thoughts of helplessness and a decrease in marital distress. By the end of therapy participants practiced timed intercourse more reliably and reported unchanged sexual pleasure and satisfaction during the nonfertile period of the menstrual cycle. At the 6-month follow-up, problem-focused thoughts had decreased. The live birth rate was higher in the therapy group than in epidemiological samples. CONCLUSION: Preliminary data suggest that cognitive behavioral treatment may be an effective approach for the treatment of infertility. PMID- 9873238 TI - Attentional correlates of illness anxiety in a non-clinical sample. AB - BACKGROUND: Attentional processes are assumed to play an important role in the maintenance of illness anxiety, although empirical support is relatively scarce. METHODS: The present study explores the relationship between selective attention (i.e. private body consciousness and symptom reporting), intensive concentration (i.e. attentional control and sustained attention), and illness anxiety in 57 non clinical subjects. RESULTS: Zero-order and multiple correlations suggest that illness anxiety is significantly related to cognitive failures in everyday life and private body consciousness and to a lesser extent to symptom reporting. CONCLUSION: It is concluded that illness anxiety can be partly predicted from specific attentional variables. However, specific operationalizations of attentional parameters seems to determine the existence and magnitude of these relations. PMID- 9873239 TI - Relationship between stress coping and personality in patients with major depressive disorder. AB - BACKGROUND: Stress coping is defined as a behavioral or cognitive response of an individual to uncomfortable or difficult situations. It has been suggested that coping, like personality, is related to the pathology and course of mental disorders. Accordingly, we here used a clinical sample to investigate the relationships between coping strategies and personality traits. METHODS: Subjects were 60 outpatients who were in remission from major depressive disorder and who completed the Coping Inventory for Stressful Situations (CISS) and the Munich Personality Test (MPT). RESULTS: Task-oriented coping showed a positive correlation with extraversion and frustration tolerance. Emotion-oriented coping was closely associated with neuroticism, esoteric tendencies and isolation tendency. Avoidance-oriented coping was related to extraversion. Principal component analysis indicated three corresponding factors between coping and personality; one was related to psychopathology (loading from the neuroticism, esoteric tendencies and isolation tendency scales of the MPT, and from the emotion-oriented coping scale of the CISS), a second was a social-adaptive ability component (loading from the frustration tolerance and extraversion scales of the MPT, and from the task-oriented coping and avoidance-oriented coping scales of the CISS), and a third was a passive-avoidance coping component (loaded from the emotion-oriented coping and avoidance-oriented coping scales of the CISS only). CONCLUSION: Some personality traits such as extraversion and frustration tolerance are significantly related to task-oriented coping, and psychopathological personality traits such as neuroticism are associated with emotional-oriented coping in major depressive disorder. PMID- 9873240 TI - Illness behavior, emotional stress and psychosocial factors among asymptomatic HIV-infected patients. AB - BACKGROUND: Over the last years the way in which patients with chronic physical illness respond to their illness (illness behavior) has been explored by several studies. This study sought to examine characteristics of illness behavior and to investigate the association between illness behavior and psychosocial and clinical variables among asymptomatic HIV-infected subjects. METHODS: Seventy three asymptomatic HIV+ outpatients completed self-report questionnaires to evaluate illness behavior (Illness Behavior Questionnaire), psychological stress symptoms (Brief Symptom Inventory), personality variables (External Locus of Control and Courtauld Emotional Control Scales) and social support (Social Provision Scale). RESULTS: Psychological morbidity ('caseness' = 34%) was associated with a pattern of illness behavior characterized by conviction of disease progression, irritability, dysphoria, psychological perception of illness and low denial. Individual capacity to express emotions, adequate levels of social support and low levels of depression, as well as clinical variables (high number of CD4+ cells, recent notification of HIV infection and nonintravenous drug use category) influenced a more adaptive illness behavior. Psychological stress and low CD4+ cell count were the main predictors of the affective dimension of illness behavior. CONCLUSIONS: Psychosocial variables resulted to influence the tendency to interpret illness in a nonadaptive way in asymptomatic HIV-infected subjects. Such variables merit to be routinely examined within the doctor-patient relationship in AIDS clinics. PMID- 9873241 TI - Infants with atopic dermatitis: maternal hopelessness, child-rearing attitudes and perceived infant temperament. AB - BACKGROUND: Atopic dermatitis (AD) is a common disease of childhood. It frequently starts in the first year of life. There is agreement on the existence of psychological influences on this disease. Although some studies in this field examine aspects of the parent-child relationship, studies concerning early infancy are very rare. The present study was conducted in order to find out whether maternal characteristics relevant to the mother-infant relationship, i.e. depressiveness/hopelessness, child-rearing attitudes and perceived infant behaviour, associated with infant AD. METHODS: Two cohorts (3- to 4-month- and 10 to 12-month-old infants), each with 20 infants suffering from AD, and 20 healthy infants were recruited. AD infants were further divided into subgroups according to the diagnostic criteria: atopic family history, itching and characteristic locations of eczema. After a paediatric examination of the infant, mothers completed standardized questionnaires concerning depressiveness/hopelessness, child-rearing attitudes and perception of infant behaviour. RESULTS: Varying with different diagnostic features of the infants' AD, mothers of AD infants described themselves as more depressive/hopeless, as more anxious/overprotective and characterized their infant as less frequently positive and more frequently negative in its emotional behaviour compared to the control group. CONCLUSION: The results underline the importance of psychological support for mothers of infants with AD. PMID- 9873242 TI - A preventive intervention program in adolescent schoolgirls: a longitudinal study. AB - BACKGROUND: To explore the impact of a prevention program on the eating and body attitudes of a sample of adolescent schoolgirls. METHODS: The program involved lessons and group discussions of general adolescent problems and eating disorders. A total of 254 16-year-old schoolgirls were evaluated, of whom 154 participated in the program and a further 154 subjects formed the control group. Variations in weight, Eating Attitudes Test and Eating Disorders Inventory at a 1 year follow-up were compared for the two groups. RESULTS: Among high-risk subjects, no significant differences were found between the prevention and the control group. The preventive program appeared to reduce significantly body dissatisfaction and to decrease the risk of bulimic attitudes in low-risk subjects. CONCLUSIONS: Providing schoolgirls with the correct information about eating disorders did not encourage unhealthy attitudes to eating and weight regulation practices. However, for high-risk subjects more intensive and specific intervention may be required, for which further research is needed. PMID- 9873243 TI - Eating problems and related weight control behaviour in adult Japanese women. AB - BACKGROUND: Fewer studies concerning eating problems have been conducted in adult than in adolescent female populations. The aims of this study are to ascertain the proportion of eating problems and clarify weight control behaviour in adult Japanese women. METHODS: This study employed a questionnaire survey with a cross sectional design. Subjects were adult females aged 20-39 years, working in a computer factory. Four hundred and six women agreed to participate in the study. The questionnaire elicited self-reported age, body height, body weight and various histories of dieting, fasting and purging behaviour. The Eating Attitudes Test 26-item version (EAT-26) was also administered. Subjects showing an EAT-26 score equal to or greater than 20 were identified as having eating problems. RESULTS: Among the subjects, 172 women (42.4%) had some history of dieting behaviour and 24 (5.9%) had some history of fasting behaviour. Regarding misuse of laxatives, diet pills and diuretics for the purpose of weight loss, 50 (14.3%) were past or current users of diet pills, followed by 42 (10.3%) laxative abusers and 15 (3.7%) diuretic abusers. Six (1.5%) subjects had an EAT-26 score of 20 or greater, 5 (1.9%) in their 20s, and 1 (0.8%) in her 30s, the difference not being statistically significant. CONCLUSIONS: Although dieting behaviour is common in adult Japanese working women, the proportion of eating problems is low in comparison with Western adult populations. PMID- 9873244 TI - Phylogeny of Tamm-Horsfall protein. AB - The Tamm-Horsfall protein (THP, uromodulin) is urine's most abundant protein. The phylogenetic distribution of the gene for THP was examined in all the major vertebrate classes. Genomic DNA was obtaind from each vertebrate class and hybridized with a rat THP cDNA probe in a DNA hybridization assay. In separate experiments, a polymerase chain reaction assay was used to amplify segments of the THP gene using primers from a consensus sequence for THP among multiple mammalian species. Both methods revealed that the THP gene was present in all the vertebrate classes. These data demonstrate the evolutionary conservation of the THP gene among vertebrates and suggest a role for this protein that is common to the kidney of all vertebrates. PMID- 9873245 TI - Reappraisal of the quantity and nature of renal calcifications and mineral metabolism in the magnesium-deficient rat. Effects of treatment with potassium citrate or the combination magnesium citrate and potassium citrate. AB - There is an urgent need for drugs capable of inhibiting renal calcifications, nephrocalcinosis and stones included, in humans. Current anticalcification medication is based mainly on alkalinization of the metabolism using potassium containing citrate alone, despite the fact that calcium stone patients suffer marginally from both magnesium and potassium deficiency. We investigated the anticalcification efficacy of oral potassium citrate versus the combined administration of this drug and magnesium citrate in the magnesium-deficient rat developing corticomedullary nephrocalcinosis and luminal microliths in the long term. Among other things we employed specific stains for calcium and oxalate, light microscopy and element analysis for renal tissue and calcifications, respectively. In addition, minerals in renal tissue, urine and plasma were determined, as well as the state of extracellular calcium homeostasis. Magnesium deficiency caused pure calcium phosphate tissue deposits, containing no magnesium, but no deposition of calcium oxalate in the tubular lumen; tissue magnesium, calcium and phosphorus were increased, and there was marked potassium wastage via urine; despite mild hypercalcemia other signs of hyperparathyroidism were not found. Alkalinization with the two kinds of medication evoked an increase in urinary pH, citrate, and potassium; however, potassium citrate alone tended to aggravate renal concretions, whereas the combination of this drug with magnesium citrate completely prevented concretions. It was concluded that: (1) magnesium deficiency-induced calcifications are oxalate-free and are not sensitive to mobilization by alkalinization with potassium citrate, which might explain the failure of the drug to prevent stone recurrence in clinical stone patients, and (2) the combination of potassium citrate and magnesium citrate, which shows enormous anticalcification efficacy, deserves high priority in clinical trials aimed at evaluating strategies for the prevention of stones. PMID- 9873246 TI - The long-term outcome of cystinuria in Japan. AB - Recurrence of growth of urinary stone is frequently observed during the clinical course of cystinuria patients. The aim of the present study is to examine the long-term outcome of cystinuria in Japan and clarify the effects of medical treatment on urinary stone. Thirty-one patients with cystinuria who had been followed up longer than 6 months were included. The follow-up period was 6-264 months with a mean of 89.5 months. Stone event was defined as appearance of new stone or radiological evidence of stone growth. All patients were managed with forced hydration and urine alkalization. Twenty-eight patients were treated with administration of thiol such as D-penicillamine or alpha mercaptopropionylglycine. Stone events per year ranged from 0 to 1.09 with a median of 0.09. Stone events per patient-year was 0.19 for all patients. The average urinary cystine concentration during treatment in the favorable outcome group (stone events per year < 0.3) was lower that that in the unfavorable outcome group (stone events per year >/=0.3); 221.2 +/- 75.2 vs. 303.3 +/- 93.5 mg/l, although the difference was not statistically significant. Prognosis of urinary stone in Japanese patients with cystinuria was relatively good with large variation. The medical treatment to reduce urinary cystine concentration would be useful for the management of cystinuria. PMID- 9873247 TI - A clinicopathological study of p53, p21 (WAF1/CIP1) and cyclin D1 expression in human prostate cancers. AB - A series of 66 prostate cancer samples were studied immunohistochemically for expression of p53, p21, a cyclin-dependent kinase inhibitor regulated by p53, and cyclin D1, a cell cycle regulatory protein. Twenty samples (30%) showed positive staining for p53, 14 (21%) for p21, and 20 (30%) for cyclin D1. p53 expression was correlated with a high Gleason score while p21 and cyclin D1 did not demonstrate any clear clinicopathological factors. A positive correlation between p53 and p21 expression was observed, however these samples with both positive immunoreactivity had no p53 mutation, suggesting the possibility that p53 may be wild-type and induce p21 expression, and/or p21 is likely to be induced by a p53 independent pathway. PMID- 9873248 TI - Study on the origin of seminal leucocytes using split ejaculate technique and the effect of leucocytospermia on sperm characteristics. AB - The origin of seminal leucocytes and their biological significance were investigated in 76 whole ejaculate samples and 27 split ejaculate samples, obtained from patients attending the Zimbabwe Family Planning Council's Spilhaus Infertility Clinic at Harare. The leucocytes were more prevalent in fractions 1 and 2 than in fraction 3, implying that the testis, epididymis and prostate are the major sources of seminal leucocytes. The contribution from the seminal vesicles was minimal. An inverse relation is apparent between leucocyte count and sperm count (p < 0.01). The percentage of abnormal sperms was higher (p < 0.05) and the sperm motility poorer in leucocytospermic samples (p < 0.01). Fructose, the seminal vesicular marker, citric acid, the prostatic marker and alpha glucosidase, the epididymal marker were not decreased in leucocytospermia. It is concluded that the epididymis and prostate are the major contributors of granulocytes in semen. Leucocytospermia affects sperm morphology and sperm motility but not the accessory sex gland functions. Probably these cytotoxic effects are mediated by hydrogen peroxide due to activation of seminal leucocytes. However, the presence of leucocytospermia in normozoospermic samples is indicative of the possible peaceful coexistence of leucocytes and sperms. PMID- 9873249 TI - Retroperitoneoscopic partial adrenalectomy using an endoscopic stapling device in patients with adrenal tumor. AB - We performed 15 retroperitoneoscopic partial adrenalectomies using an endoscopic stapling device to divide the normal adrenal tissue and adenoma. The mean operating time and mean blood loss were 162 (129-210) min and 12 (5-67) ml, respectively. In 1 patient (6.7%), a pneumothorax occurred at the end of the procedure due to injury of the medial crus of the diaphragm. However, the procedure was not subsequently converted to open surgery. These facts suggest that posterior retroperitoneoscopic partial adrenalectomy is a valuable addition to adrenal gland surgery. PMID- 9873250 TI - Transurethral needle ablation of the prostate: an alternative minimally invasive therapeutic concept in the treatment of benign prostate hyperplasia. AB - Alternative minimally invasive methods treating benign prostate hyperplasia (BPH) have become more and more important. Transurethral needle ablation (TUNATM) has been demonstrated to be effective in both canine and the human prostate. The goal of our study was to prove the safety, feasibility and tolerance of this new procedure. In this prospective nonrandomized study, 33 patients underwent TUNATM treatment. One, 3 and 6 months postoperatively, follow-up examinations were carried out assessing urodynamic parameters such as urinary flow rates, residual urine levels, and IPSS-scores. During the last visit additional cystomanometry and urethrocystoscopy were done. Six months postoperatively IPSS-score and residual urine volumes were decreased by 50% (range: 1-72%) and 75% (range: 12 97%). Improvement in maximum flow rate was 63% (range: 5-125%). No serious postoperative complications occurred. After a short 'learning period' most of the treatments where performed as an out-patient-procedure so the patients could leave the hospital without the need for indwelling catheters. The TUNATM appears to be a minimally invasive and safe out-patient procedure for the treatment of selected cases of BPH. PMID- 9873251 TI - Infertility and multiple urogenital abnormalities in a male with mosaic 46,XY/45,XO/47,XXY karyotype and mixed phenotype. AB - We hereby present a rare case of a 46,XY/45,XO/47,XXY mosaic male patient with a predominance of the XY cell line. The patient, who exhibited phenotypic stigmata of both XO gonadal dysgenesis and Klinefelter syndromes, suffered from infertility and multiple urogenital abnormalities, as our investigation revealed. PMID- 9873252 TI - Acute onset of coagulopathy in a patient with Kasabach-Merritt syndrome following transurethral resection of bladder tumor. AB - We report an unusual case of Kasabach-Merritt syndrome in a 62 year-old woman with bladder tumor who suffered from acute consumption coagulopathy that increased the fibrinolytic activity due to the presence of a huge hemangioma in the upper and lower extremities. Two days after the transurethral resection of the solitary bladder tumor, serious hematuria and life-threatening disseminated intravascular coagulation developed. Following treatments with heparin and subsequent steroid treatment, hemorrhage and abnormal hemostatic values improved. It is suggested that extensive investigation of the coagulation system should be done in patients with giant hemangioma, particularly before a surgical procedure. PMID- 9873253 TI - Testicular teratoma with a unique mode of spread and spontaneous maturation. AB - Chylous ascites, a rare compliation following retroperitoneal lymph node dissection, has not been reported as a means of spread of testicular cancer. This report describes a unique path of spread of testicular cancer which also appears to spontaneously mature at the metastatic site. PMID- 9873254 TI - Giant hydronephrosis--a diagnostic dilemma. AB - We report a complex case in which the left kidney had undergone giant hydronephrotic change after chronic obstruction at the vesicoureteric junction. Minor blunt abdominal trauma caused rupture of the parenchyma of this expanded and dilated kidney, with bleeding into its collecting system. The mixture of blood and urine remained contained within the kidney's structural layers, so producing a tense, cystic, fluid-filled mass arising from the left hypochondrium. Pathogenesis, differential diagnosis and investigation of giant hydronephrosis and its rupture are discussed. The observation is made that gross distortion of the renal parenchyma by rupture or hydronephrosis impairs arterial inflow to the kidney. PMID- 9873255 TI - An unusual filling defect in the ureter. AB - A case of benign ureteric haemangioma is reported. A conservative approach is recommended when the kidney is capable of function. PMID- 9873256 TI - Solitary cutaneous metastasis of superficial bladder cancer. AB - Cutaneous metastases from bladder cancer are an uncommon finding. We present a patient with superficial bladder cancer whose first manifestation of disseminated disease was a solitary cutaneous metastasis of the scrotum. PMID- 9873257 TI - Eosinophilic cystitis masquerading as invasive bladder cancer. AB - We report the case of a 33-year-old woman with eosinophilic cystitis, which masqueraded as invasive bladder cancer. The patient remained well with no recurrence or progression after vigorous transurethral resection of the lesion. PMID- 9873258 TI - Successful closure of neovesicocutaneous fistula with fibrin glue. AB - We report a case of neovesicocutaneous fistula following radical cystectomy with urinary diversion via an ileal neobladder. The fistula was closed with fibrin glue. The present case suggests that occlusion therapy with fibrin glue is useful as a treatment option to facilitate the closure of a fistulous tract in the case of prolonged urine leakage from the ileal neobladder. PMID- 9873259 TI - Ureteroscopic approach to early postoperative ureteral obstruction in the renal transplant patient. AB - The ureteroscopic treatment of acute postoperative ureteral obstruction in a kidney transplant patient is presented. This approach was made possible by the use of thin instruments, which do not require predilation, and was chosen in place of antegrade nephrostomy or open surgical access. The concept that the ureteroscope must be adapted to the ureter and its characteristics and not vice versa is stressed. The increasingly widespread availability and use of thin and ultrathin ureteroscopes will ensure that this approach is likely to become the rule rather than the exception in the future. PMID- 9873260 TI - Progression to AIDS in relation to clinical factors and clotting product consumption. A 14-year follow-up of a cohort of 52 Dutch HIV-1-infected haemophilic patients. AB - OBJECTIVE: In order to describe the progression of HIV disease in a cohort of Dutch HIV-1-infected patients with haemophilia and to study the influence of clinical and clotting factors, we performed a 15-year longitudinal clinical and laboratory study. METHODS: The progression to AIDS in relation to type of haemophilia, antibodies against factor VIII, age, cytomegalovirus (CMV) infection, and type and total amount of clotting factor consumption was studied in 52 haemophilic patients infected with HIV-1 between 1981 and July 1985. RESULTS: The progression to AIDS was faster in the group of patients with antibodies against factor VIII (inhibitors) and in the group of patients with haemophilia B than in the group of patients with haemophilia A without inhibitors. CONCLUSION: We concluded that HIV-infected patients with haemophilia A with inhibitors and patients with haemophilia B show a significantly faster progression to AIDS than do HIV-infected patients with haemophilia A without inhibitors. These differences are independent of the total amount of clotting factor used and other known risk factors such as age at seroconversion and CMV infection and may be due to the type of clotting product used. PMID- 9873261 TI - Hepatitis E virus: relevance in blood donors and other risk groups. AB - BACKGROUND AND OBJECTIVES: Hepatitis E virus (HEV) infection usually causes an acute self-limited disease. HEV is associated with feces-contaminated drinking water, but other vectors, such as blood, are possible. The aim of this study was to investigate the prevalence of HEV in blood donors and in two groups at high risk of parenteral infections, namely, hemodialysis patients and children infected with hepatitis C virus (HCV) via blood transfusion. MATERIALS AND METHODS: We investigated the prevalence of anti-HEV in 863 blood donors, 63 hemodialysis patients, and 42 children infected with HCV posttransfusion. RESULTS: The prevalence rates were 2.8, 6. 3%, and 0 respectively. CONCLUSIONS: (1) The incidence of HEV in Spain is similar to that in other western European countries, and (2) HEV is probably not transmitted parenterally to children. PMID- 9873262 TI - A method for the quantitative assessment of platelet-induced clot retraction and clot strength in fresh and stored platelets. AB - BACKGROUND AND OBJECTIVES: The changes that occur in platelets as they undergo storage have been documented by aggregometry as well as by flow cytometry. However, one of the most essential platelet functions, the induction of clot retraction, has not been quantitatively assessed in stored platelets. We describe two potentially useful methods, platelet-induced clot retraction and clot strength, to assess effect of storage of platelets in blood banks or of platelet preparations subjected to freezing or freeze-drying. These methods have previously been developed for bedside monitoring of patients receiving c7E3 (Reopro(R)). MATERIALS AND METHODS: Platelet-induced clot retraction (PICR) and clot strength were measured with the Hemodyne and Thromboelastograph, respectively. Paired Study: Fresh platelet concentrates (n = 3) were obtained from leukapheresis donors and divided into two equal units; one unit was tested within 4 h of collection and the other stored for 5 days at 22 degrees C in a platelet incubator and tested. Unpaired Study: Fresh platelet concentrates (n = 15) were obtained from leukapheresis donors and tested within 4 h of collection and compared to outdated platelets (n = 30; random or single donor) that had been stored for 5 days at 22 degrees C in a platelet incubator. Alternative Preservation Methods: Lyophilized platelets, platelets chilled to 4 degrees C, platelets frozen at -70 degrees C in 5% dimethyl sulfoxide (DMSO) or in the absence of a cryoprotectant. RESULTS: Paired Study: Stored platelets demonstrated an increase in PICR; the difference was not significant (p = 0.55). There was no difference in clot strength between fresh and outdated platelets (p = 0.90). Unpaired Study: When compared to fresh platelets, stored platelets demonstrated a 2-fold higher PICR (p = 0.0011). On the other hand, there was no difference in the time to onset of PICR (p = 0.08) and there was no difference in clot strength between fresh and outdated platelets (p = 0.14). Alternate Preservation Methods: In contrast, PICR and clot strength were reduced in platelets frozen at -70 degrees C in 5% DMSO and absent in lyophilized platelets, in platelets frozen at 70 degrees C in the absence of cryoprotectants or stored at 4 degrees C. CONCLUSION: The data indicate that the ability of platelets to induce clot retraction and to enhance clot strength is not altered by storage, despite functional abnormalities in aggregation and agglutination. These data suggest that quantitative measurements of PICR and clot strength may be simple, useful tools for assessing the function of stored platelet concentrates, platelets that have undergone freezing or exposure to alternative buffers and for evaluating platelet functions relevant to PICR. PMID- 9873263 TI - Sequence and specificity analysis of recombinant human Fab anti-Rh D isolated by phage display. AB - BACKGROUND AND OBJECTIVES: Hyperimmune anti-Rh D serum is worldwide in short supply. As a first step to develop an alternative source of Rh D antibodies, we describe in this study the isolation and characterization of recombinant anti-Rh D Fab fragments. MATERIALS AND METHODS: Peripheral blood mononuclear cells harvested from a hyperimmunized donor were used to construct two combinatorial Fab libraries. Phages expressing these Fab fragments were selected on whole red blood cells followed by testing of positive clones in an indirect hemagglutination assay. RESULTS: Individual Fab clones are of high affinity and competitively inhibit the binding of a registered anti-D immunoglobulin. The Fab clones are also specific against the partial D phenotypes, Rh33, DIII, DIVa, DIVb, DVa, and DVII. The 13 different but highly homologous clones express preferentially VH3 segments. CONCLUSION: These Fab fragments show potential for the development of a new generation of therapeutic anti-Rh D reagents. PMID- 9873264 TI - A stable reagent system for screening and identifying red blood cell irregular antibodies: application to commercial antibodies. AB - OBJECTIVE: Development of a new solid-phase system for screening and identifying irregular red cell antibodies. MATERIALS AND METHODS: Red blood cell membranes were prepared by a semi-automated procedure in which the hemolysate solution was passed through a hollow-fiber system. The membranes were fixed to the solid phase (microtiter plates) by centrifugation and incubated with 8% fat-free milk. Antibodies added to the microtiter plate were detected by anti-human antibodies adsorbed onto yellow latex particles. RESULTS: The system had good sensitivity (titer <1); 97% of anti-D samples were detected. The detection system was stable for 6 months at 4 degrees C. CONCLUSION: This stable-antigen solid-phase system readily detects and identifies red cell antibodies that are important in transfusion. PMID- 9873265 TI - Detection of HIV-1 RNA in two consecutive blood donations screened negative for HIV antibodies. AB - BACKGROUND AND OBJECTIVES: An unusual serological pattern of HIV-1 seroconversion in a blood donor is described. The seroconversion panel was used to investigate the sensitivity of existing screening assays. MATERIALS AND METHODS: A volunteer blood donor who had given blood 79 times was diagnosed anti-HIV-1-antibody positive. The heteroduplex mobility assay identified a subtype B HIV-1 strain. The frozen plasmas from the last four blood donations had been kept at -30 degrees C. They were thawed and aliquoted for subsequent testing. RESULTS: The last two blood donations contained HIV-1 RNA, 2,800 copies/ml (October 26) and 170 copies/ml (November 23). Weak anti-p24 antibodies were detected by Western blot in the October 26 sample, and a clear p24 reactivity along with a faint gp160 reactivity was observed on November 23. HIV p24 antigen was undetectable in both samples. Out of 13 screening assays, only 6 gave positive results on the November sample and 7 negative results which were obtained by 1 competitive enzyme immunoassay (EIA) and 6 of the 9 sandwich EIAs. CONCLUSION: Most sandwich EIAs gave prolonged false-negative results in the present case. p24 antigen testing was negative and would not have reduced the risk of HIV transmission. PMID- 9873266 TI - Detection of HIV-1 infection in dried blood spots from a 12-year-old ABO bedside test card. AB - BACKGROUND AND OBJECTIVES: We tested dried blood from an ABO bedside test card which had been stored at room temperature for 12 years, to prove that a patient with HIV-1 infection had been infected by blood transfusion. MATERIALS AND METHODS: Immunoblots for HIV-1 antibodies and threefold PCRs with half-nested primers for the HIV-1 integrase gene were done with eluates from the dried blood spots. RESULTS: HIV-1 antibodies and HIV-1 DNA could be detected in the sample from one unit of blood, but not from the two other units or from the recipient before transfusion. CONCLUSION: Further studies should be done on the validity of stored dried blood as an alternative to the storage of frozen donor serum for several years for 'look-back' studies. PMID- 9873267 TI - Potential usefulness of photochemotherapy using 8-methoxypsoralen in the treatment of posttransfusion graft-versus-host disease. PMID- 9873268 TI - The official requirements for platelet concentrates. PMID- 9873269 TI - Another piece of the patient history puzzle. PMID- 9873270 TI - Photosensitivity: classification, diagnosis, and treatment. AB - Photosensitivity disorders are common. Phototoxicity due to excess ultraviolet exposure while on a photosensitizing drug and polymorphic light eruption are two frequent examples. Patients should be encouraged to consider Taylor's ABCDEs: Avoidance, Barrier creams, Clothing, Dihydroxyacetone, and Education. PMID- 9873271 TI - High-dose adjuvant interferon therapy for melanoma. AB - High-dose interferon has recently been approved as an adjuvant to surgery for treating newly diagnosed patients at high-risk for melanoma, particularly those with regional lymph node disease. The data supporting the indications for treatment and its side effects are reviewed. PMID- 9873272 TI - What's your assessment? Acne rosacea. AB - The "What's Your Assessment?" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 9873273 TI - Obtaining the optimal treatment outcome with tazarotene. AB - Tazarotene topical gel (Tazorac) is a vitamin A derivative and the first topical retinoid to demonstrate therapeutic success in treating plaque-type psoriasis. It has been available in the United States since 1997. Tazarotene offers long-term remission in many patients and may be used in combination with corticosteroids and UVB therapy for even better results. Due to its potency as a vitamin derivative, tazarotene is also associated with irritation in some patients. Patients must be educated about proper application of the drug and realistic expectations for treatment outcome. PMID- 9873274 TI - Complications of cutaneous laser resurfacing: a nursing guide. AB - Laser use for various cutaneous conditions has advanced rapidly in the past several years. Initial background and physical history is important when determining if a patient is a candidate for laser resurfacing. Nurses must be able not only to identify appropriate candidates preoperatively, but to discuss the risks associated with the procedure. Because proper postoperative management is critical to the healing process, nurses must identify complications early in their course to provide appropriate intervention and, thus, reduce long-term side effects. PMID- 9873275 TI - Developing and evaluating clinical supervision in the United Kingdom. AB - This paper provides an overview of the definitions and meaning given to clinical supervision in the United Kingdom. In addition, reference and examples from evaluation research undertaken by the University of Manchester are given which offers insight into the content, focus and purpose of clinical supervision in a variety of clinical specialities. Comment is offered on the value of research and the limitations and possibilities for research methods in evaluating clinical supervision. PMID- 9873276 TI - Job description for the Certified Nephrology Nurse: professional and legal aspects. AB - As in many European countries, in Germany, practice and responsibilities of the Certified Nephrology Nurse (CNN) are still affected by a slowly changing structure in health care-providing institutions and the developing qualification process of the nurse working in the renal field. It is evident that parallel to the development and implementation of the European Core Curriculum the scope of practice for the CNN needed to be defined and published by a group of clinical experts working in the renal nursing field. A task force composed of members of the EDTNA-ERCA and the German National Renal Care Association was set up to tackle all aspects of the professional profile. A lawyer experienced and recognised in the medical field supported the group. In this paper information about the background and content of the document is presented, as are the core areas of practice and responsibilities and areas where junior nurses or helpers may work under the guidance of CNN, also the professional and legal aspects of the CNN profile are addressed, discussing possible benefits and problems for colleagues in practice when using and implementing the documents in different structures of renal care-providing organisations. This document was published and sent to all important national Bodies in Germany, having an immediate impact in units where areas of practice and responsibilities were undefined for certified renal nurses and nurses auxiliaries. PMID- 9873277 TI - Patient classification used as a tool for assessment of staff/patient ratios. AB - Within the bounds of health care reforms nephrology nurses have a joint responsibility to make costs transparent and to decrease them. Care for chronically ill patients must constantly be delivered and there should be no limitations and changes in nursing and medical care. With the establishment of a clear job description and a performance profile for nephrology nurses a reorganisation of staffing is possible. The employment of non-qualified staff for indirect and parts of general patient care is possible as long as the work is done under the supervision and responsibility of a registered qualified nephrology nurse. With patient classification it is clear how much and which kind of care has to be provided and who should carry the costs. Furthermore documentation of actually needed care is possible. The further qualification of nurses with a post basic course for nephrology enables us to work with non qualified staff and contributes to a high qualified standard of care. With regard to material used for dialysis costs should be transparent and nurses should be aware of them. The amount of unnecessarily used materials should be reduced and working process reviewed and economically organised. There are some aspects to be considered, e.g. the use of sterilised material for work which could be done with non-sterilised material which is much cheaper. We should have in mind that reimbursement rate combines costs for material and staff and if we could reduce the costs for used material there is more money left to pay the nurses. Last but not least all dialysis is providers should work closer together and should avoid confrontation. Confrontation means counterproductive and this is expensive. Only a concerted action of everybody involved in renal care may contribute to ensure the current standard of renal replacement therapy. The awareness of nurses should be developed that they contribute to safe their own place of employment. PMID- 9873278 TI - Informed consent: do we achieve it? What are the elements that help create it? AB - Informed consent is a complex concept, it raises issues which involve ethical, moral and legal debates. This paper is intended to explore the elements of informed consent and to give guidance to those in the process of gaining consent from patients. The process should be seen as a partnership between the multidisciplinary team and the patient. PMID- 9873279 TI - Understanding other cultures: the need for training. AB - People from ethnic minorities from a substantial group treated in the renal unit, some 10% or more. Because of the language, dietary and cultural differences, there are special problems for the multi-disciplinary team in communication, understanding and sensitivity to the particular needs that may arise. Three case studies and their implications will be discussed. In all these situations staff felt helpless and de-skilled, having no knowledge of the cultural implications. Questionnaires show that the majority of staff from all disciplines receive very little, if any, training in the faiths and cultural traditions of those from other countries--a situation that needs urgent attention. PMID- 9873280 TI - Care of the older person with kidney failure. AB - Dialysis and renal transplants have become a reality in the XXIst century. Their use in third world countries has profound impacts on social, economic, political and cultural aspects. Extraordinary achievements for the medical world, that is constantly concerned in improving the health levels for the population; but what are the repercussions for the lower income groups? Is the cost/benefit relationship being analysed? Is there ample information available and is it well distributed regarding how to avoid the need for dialysis or a kidney transplant? What recommendations are made to the health care workers? And, what has been revealed to us after reflecting on the philosophical/historical aspects of the human existence? How valuable are Dr. Ivan Illich's observations and what can we expect from health care workers in relation to such significant achievements as we face the XXIst century? PMID- 9873281 TI - Making decisions about dialysis options: an audit of patients' views. AB - This paper describes the evaluation and audit of 297 patients referred for pre dialysis education. Education and support for patients include home visits, visits to the dialysis units, written, visual and audio educational material and patient information day. There was a 50-50 split between patients opting for haemodialysis and peritoneal, while in circumstances where patients presented acutely or at end-stage, the split was 80-20 in favour of haemodialysis. Patients views were sought by postal questionnaire (Over 75% of patient satisfaction with information), areas of least satisfaction concerned adaptation to everyday life on dialysis, and potential problems and complications of treatment. Evaluation has been favourable. Recommendations include changes in the format and management of the education programme and in particular with regard to patients that present at end-stage. PMID- 9873282 TI - Patient adherence in renal replacement therapy: whose problem is it? The physician's perspective. AB - Patient compliance is an issue sadly neglected in the literature. Out of 470 scientific papers concerning dialysis published in 1996 only 4 (< 1%) addressed the subject. Doctors have an important role in the process leading to patient adherence, as they often see the patient on an outpatient basis for years before the start of dialysis and have an opportunity to prepare the patients for the forthcoming demands. Studies have shown that patients not seeing a Nephrologist before start of dialysis have a higher mortality and morbidity rate. The doctor has also an important role when informing the patient about the consequences of non-compliance. The aim should be to make the patient feel that he or she is a valuable member of the caring team. PMID- 9873283 TI - Psychological nursing support for elderly patients undergoing chronic regular haemodialysis. AB - The elderly constitute a continuously increasing social group of the Hellenic, but also of the global population. This phenomenon is also evident in the haemodialysis patient population, which grows continually. Faced with this reality, Nursing is obliged to adapt itself and, in this effort that it is putting forward it has achieved many positive steps (geriatric nursing). It is imperative however for nursing that there is a particular way of dealing with the elderly, especially those that undergo chronic periodic haemodialysis, which regards as much the problems stemming from the disease, as it regards their grave psychological condition. The objective of our study was to develop the need for the presence of the Nephrology Nurse in the psychological support of the elderly renal patient and we have been able to define: level of communication, level of dietetic information, psycho-social condition and restrictions imposed by the disease. 30 patients (age 65-80, mean 72.92 years) were included with an observation time of 3 years, less than 60% responded positively to the efforts for psychological support, on the basis of the studied factors. The rest did not show any willingness to answer. We conclude that psychological support is of paramount importance for these patients but also that the Nephrology Nurse has not yet discovered and detected some unknown aspects of the different problems arising due to insufficient knowledge of geriatric caring. PMID- 9873284 TI - Pre-dialysis information: how effective is it? AB - Many individual with decreased renal function are reviewed in pre-dialysis outpatient clinic for a varying length of time. It is in the pre-dialysis stage that the physician informs the individual about their progressive kidney failure and of the eventual need for dialysis. Despite the length of time that the patient has prior to commencing a renal replacement programme the renal team attempt to prepare the patient for their impending treatment by providing adequate information and allowing them to explore their illness and their future. Research has shown that the more knowledge the patients have about their disease and subsequent treatment, the more they are able to participate in their own care and the better they feel, both mentally and physically. All the patients receiving pre-dialysis information in the last 6 months are included, 14 subjects were chosen at random to participate in the study. A standard information-giving service has now been developed within the renal service, based on the needs of the patients using the service. PMID- 9873285 TI - "Non-compliance": meaningful construct or destructive, sticky, stigmatizing label? AB - The "(non-) compliance" is critically reviewed in term of both published research and clinical experience. Underlying assumptions are examined, and assessment problems and relevant theoretical models briefly reviewed. The Self-Regulation Model is outlined in some detail as a basis for both clinical interventions and research. Attention is drawn to the potential for prejudice and stereotyping as a result of the application of a negative evaluative label like "non-compliance". It is concluded that "(non-) compliance" is a mythical construct and proposed that the evaluatively neutral, non-judgmental, term self-management behaviours be substituted. Directions for further research are outlined. PMID- 9873286 TI - "Lose not an hour": patient decision-making and improving quality services through the prism of renal care. AB - This paper will discuss the scarcest resource in the world--the days of people's lives. And how to help all to live to the fullest. There is some room, of course, for patients and professionals to disagree. Sometimes there might be too much emphasis on the professional view. Disagree? Well, we should remember that though the "Titanic" was built by professionals, the "Ark" was built by amateurs! Society needs to find new answers for empowering patient decision making about the quantity, quality and access to care. Without changes in financial structures there is little prospect of empowering patients to make decisions for themselves and empowering professionals to do the job as they hope to do it. PMID- 9873287 TI - Dialysis time: what you prescribe is what you get? (WYPIWYG). AB - The duration of a dialysis session is an essential parameter in determining dialysis efficiency and hence adequacy. The aim of this study was to measure the real dialysis time and to compare it to the prescribed time. For 12 months, 8412 dialysis treatments were allocated a number and a random sample of 100 was selected using a computer generated randomisation code. At the end of each dialysis, the time prescribed, the real time achieved, the blood volume exchanged, and any incidents occurring during the dialysis were noted. All the staff was aware of the study but nobody except the controller knew which dialysis was going to be controlled. The study population consisted of 48 patients, 25 males and 23 females. The mean duration of the prescribed dialysis time was 3.30 hours. Three patients used the same monitor per day. Results showed a loss of 10 minutes for 180 minutes dialysis prescription, 9.5 minutes for 210 minutes and 10.5 mintutes for 240-dialysis prescription. The dialyses with the greatest difference in time measured and prescribed occurred on the second (12 am to 3 am) shift (-6.23) which might be explained by the necessity for changing the bicarbonate cartridge during the shift. This loss of time represents a mean of 7 dialyses per year per patient. In conclusion, the prescribed time for a dialysis is very often not respected. A readjustment of the time during the treatment will be necessary to compensate for this loss of diffusion time and consequent efficiency. PMID- 9873288 TI - Clueing in: a guide to solving the puzzle of self for women recovering from depression. AB - Clueing in, a process constructed from data collected from women recovered or recovering from depression, seems to be a salient variable in reaching full recovery. While depression and its recovery are composed of a number of highly complex processes, when women clue in, or come to a cognitive and emotional realization of themselves in relation to their world, they seem able to put in place the final piece of the puzzle of who they are, that is, what is their true self. For women to reach full recovery, it seems necessary for them to discover the difference between what and who they are and are not responsible for in their lives. Clueing in is one component of a model of recovery developed from a grounded theory study conducted with 21 women during 1993 to 1995. PMID- 9873289 TI - Power and the politics of abuse: rethinking violence in Filipina-Australian marriages. AB - In this paper we take as a starting point the perceived high prevalence of domestic violence in marriages between Filipino women and Australian men. In in depth interviews and structured questionnaires with service providers and with Filipinas married to Anglo-Australian and Filipino men the issue of underreporting physical and emotional violence was a recurring theme. We explore the relationships of power that characterize these marriages, the changes in balance of power that result from the externalization of anger, physical and verbal violence, and stigmatization of domestic violence experienced by Filipinas individually and in the community. We suggest that social disapproval and stereotypical representations of Filipina-Australian marriages in Australian society, and the consequent shame experienced by Filipinas, has led to underreporting of emotional and physical abuse. PMID- 9873290 TI - Menarche stories: reminiscences of college students from Lithuania, Malaysia, Sudan, and the United States. AB - Women college students in four countries were invited to write the story of their first menstruation in as much detail as memory allowed. Stories were received from 26 Lithuanians, 27 Americans, 20 Malaysians, and 23 Sudanese. The stories were read and their contents analyzed for the presence or absence of information on such topics as emotional reaction, preparedness, sources of information about menstruation, changes in body image, and celebrations of this rite of passage. Similarities and differences among the groups are discussed, and passages from particularly interesting stories are quoted. PMID- 9873291 TI - Stroke risk factor knowledge in Hispanic and non-Hispanic white women in New Mexico: implications for targeted prevention strategies. AB - Hispanic women in New Mexico have recently experienced an increase in age adjusted mortality compared with non-Hispanic white women. Since patients' knowledge of stroke risk factors may affect risk factor control, the present study was undertaken to characterize stroke risk factor understanding in Hispanic and non-Hispanic white women in New Mexico. We administered a stroke risk factor knowledge survey to 215 women hospitalized in Albuquerque, New Mexico. Patients were classified by each of three dichotomous groupings: stroke or nonstroke diagnosis; Hispanic or non-Hispanic white ethnicity; history of cardiovascular risk factors. The frequency of specific item responses was determined for each patient grouping. Two-way analysis of variance was used to determine whether composite knowledge score differed among patient groups. Stress was the attribute most commonly thought to be a risk factor for stroke. Although no ethnic differences were found on composite knowledge score, Hispanic women were significantly less likely to report hypertension as a stroke risk factor than non Hispanic white women. We suggest that stroke risk factor understanding in Hispanic and non-Hispanic white women in New Mexico is inadequate. Insufficient understanding of the consequences of hypertension, including stroke, may diminish the degree of hypertension control that patients achieve. Further study of the relationship between stroke risk factor understanding and health behavior could enhance prevention efforts. PMID- 9873292 TI - Attitudes, beliefs, and practices of nursing students concerning HIV/AIDS: implications for prevention in women. AB - Nursing students are a group of predominantly young women who may be sexually active but who are well educated and presumably health conscious. It might be expected, therefore, that they are not a population at risk for sexually acquired HIV infection. Recent studies indicate that heterosexual women constitute the fastest growing population of persons with AIDS in the United States and Canada (Health and Welfare Canada, 1993b; Wofsky, 1992) and that women and adolescents will constitute the next surge of the AIDS epidemic (Novello, 1993). First-year nursing students in a major Canadian city were surveyed regarding HIV-related knowledge, attitudes, beliefs, and behaviors. The women were highly knowledgeable about HIV transmission but 15% to 25% reported high risk sexual behavior. The results reinforce that knowledge is not enough to prevent HIV infection among young women and that interventions must be based on an understanding of the social context of women's lives. PMID- 9873293 TI - The use of formal and informal health care by female adolescents in eastern Nepal. AB - I am presenting the background and interim results of an ongoing study in Eastern Nepal where at present the adolescent females do not use the formal health sector to the same extent as men. Suggested reasons for this include lack of awareness (literacy rates among women are 17%), lack of access, lack of time, and the cost and the quality of services offered. Women are more likely to visit traditional health practitioners, and very little research has been done to find out why this is the case. I also examine the access of rural adolescent females to health services and the policy process. The researcher involves the women in the research so that they can actively participate in the decision-making process and in setting health priorities. The research is based on a reconstituted form of participatory rural appraisal (PRA) known as participatory appraisal of needs and development of action (PANDA). This is a framework which has been developed out of PRA and has been used in developing countries to enable local people to obtain, share, and analyze their life conditions and to plan and act according to that knowledge. PMID- 9873294 TI - Practice development: a growing and significant movement. PMID- 9873295 TI - Ventilator technology: safety and comfort for patients? AB - Safety and comfort aspects of mechanical ventilation using advanced microprocessor ventilators are discussed. The significance of some of the ventilator observations made, both digital and waveform, is highlighted. Readers are encouraged to experience for themselves the trigger mechanisms available on the ventilators they use. This article combines theoretical information and direct reference to the Nellcor Puritan Bennett 7200 Series microprocessor ventilator, but many of the principles described can be applied to other microprocessor ventilators with flow triggers and waveform options. Nurses are encouraged to find out how this information relates to the ventilators they use. PMID- 9873296 TI - Nursing diagnosis: use and potential in critical care. AB - Nursing diagnosis is a classification system for nursing widely used in the USA. There is increasing interest in its potential for use within British nursing. Benefits and concerns are raised about using nursing diagnosis in practice. This paper considers the potential for using nursing diagnosis in a critical care setting. Using nursing diagnosis raises many issues for British nurses, and these need to be discussed and clarified prior to implementation into practice. PMID- 9873297 TI - The physiological knowledge required by nurses caring for patients with unstable angina. AB - Physiological knowledge necessary to critically analyse nursing management of patients with unstable angina is reviewed. The role of endothelium derived relaxing factor, nitrous oxide and atherosclerosis is summarised. The effects of circadian rhythms or clinical signs and symptoms in patients with unstable angina is particularly highlighted. Pharmacological interventions are considered from the perspective of implications for nursing care and other important nursing interventions identified for coronary care nurses. PMID- 9873298 TI - Advance directives, current legal and ethical issues. AB - The current legal position from which an advance directive operates is clarified. The practicalities of advance directives are considered and the issues concerning enalties explored. The patient's capacity to refuse treatment is discussed in relation to the issue of consent. Situations concerning cardio-pulmonary resuscitation, pregnancy and accident and emergency are explored. PMID- 9873299 TI - Nutritional assessment in clinical practice: a review. AB - Malnutrition is a common problem within the clinical setting. Patients affected by malnutrition are more likely to suffer serious complications with an associated increase in length of stay and cost of treatment. Despite these serious outcomes, nurses are still failing to nutritionally assess their patients. This paper explores means of determining nutritional status in order to facilitate early detection and treatment of nutritional problems. PMID- 9873300 TI - Supervision in practice. AB - The nature of the supervising relationship is explored in relation to an intensive care unit. The relationship between clinical supervision and individual performance review is discussed. Preparation and support for the supervisor's role is identified. Peer, group and external supervision arrangements are explored. PMID- 9873301 TI - Reflecting on the knowledge used when caring for an intensive care patient and his/her family. AB - Carper's four ways of knowing are used to structure a reflection on the knowledge used by an associate nurse in intensive care when caring for her patient, his wife and son. John, the patient, had previously undergone a sex change operation as well as cardiac surgery. His current period in intensive care was due to pancreatitis and involved numerous returns to theatre. He eventually died following multi-organ failure. The reflection focuses on the associate nurse's feelings when trying to act as an advocate for both John and his wife at the time of the patient's pending death. PMID- 9873302 TI - Integrated care pathways: fashionable necessity? PMID- 9873303 TI - Does the APACHE II scoring system equate with the Nottingham Patient Dependency System? Can these systems be used to determine nursing workload and skill mix? AB - The development of a holistic patient dependency system for intensive care in Nottingham has enabled nursing staff to analyse their patient's needs more comprehensively and relate them to existing staffing levels. A recent publication (Chellel et al., 1995) implied that some units were using medical scoring systems to predict workload and that practitioners were dissatisfied with current scoring systems. Patient dependency scores were correlated with APACHE II scores by regression analysis on 1,743 patients from the Adult Intensive. Care Unit at the University Hospital, Nottingham. Analysis revealed that APACHE II had little ability to predict patient dependency and, therefore, is unreliable at predicting nursing workload and skill mix. PMID- 9873304 TI - Knowledge and attitudes of nurses on a regional neurological intensive therapy unit towards brain stem death and organ donation. AB - The study aimed to explore nurses knowledge and attitudes towards brain stem death and organ donation. An ex post facto research design was used to determine relationships between variables. A 16 item questionnaire was used to collect data. Statistical analysis revealed one significant result. The limitations of the sample size is acknowledged and the conclusion suggests a larger study is required. PMID- 9873305 TI - The use of a paediatric coma scale for monitoring infants and young children with head injuries. AB - Head injuries are a significant cause of morbidity and mortality in children. A descriptive multiple-case study was used to compare the accuracy of two assessment scales in children aged under 5 years. The study emphasised the importance of prompt and accurate assessment of children with head injuries. The Glasgow Coma Scale (GCS) is inappropriate for use in young children. The Adelaide Coma Scale (ACS) is a more appropriate tool for those aged under 5 years because it takes the child's age, and, therefore, neurological immaturity, into account. However, the ACS still does not give enough scope for assessment. If the ACS is used, therefore, additional information must be considered, including specific paediatric indicators, and an assessment of each child's behaviour according to the main care giver. PMID- 9873306 TI - Reasonable refusal?--an ethical dilemma. AB - Ethical decision-making can lead to fairer and collegiate decisions, which are made collectively with the patient. This paper considers the role of the nurse in the ethical decision-making process. The experience of being involved in the decision-making process is shared. The importance of nurses becoming involved in ethical decision-making in an impartial and non-judgmental way is advocated. PMID- 9873307 TI - The metabolic changes associated with trauma and sepsis. AB - Developments in critical care medicine have increased the chances of survival of those patients with severe trauma or established sepsis. However, such patients often have a prolonged critical illness, and the ensuing catabolic response can have detrimental effects causing a depletion of body weight, tissue mass and stored nutrients. In order to facilitate treatment and minimise the effects of these catabolic changes, both the mediators of this response and the response itself have been studied. This paper explores the current understanding of the metabolic changes that occur in trauma and sepsis, and how these changes alter carbohydrate, fat and protein metabolism. PMID- 9873308 TI - Percutaneous balloon mitral valvuloplasty: advancing the nursing perspective. AB - Balloon mitral valvuloplasty is increasingly used to widen the narrowed mitral orifice in selected patients with valve stenosis and is a cheaper alternative to open heart surgery. This technique is gaining wide acceptance in clinical practice due to certain advantages, such as being less invasive and thus abolishing the need for thoracotomy, reduced costs and shorter hospital stay. Recent research indicates that there are substantial long-term benefits, such as improved functional capacity and relief from symptoms, which compare favourably with surgical valvotomy. As interventional cardiology expands, nurses must meet the new challenges and continue to expand and develop the unique nursing perspective in this field of patient care. A key role for nurses is to develop a trusting relationship, provide information to meet the patient's needs and emotional support. PMID- 9873309 TI - Empowerment. PMID- 9873310 TI - The needs of parents with a child on an adult intensive therapy unit. AB - This review examines, by means of a literature search, the needs of parents who have a critically ill child on adult intensive therapy units. These needs are compared with the needs of relatives of adult patients in ITUs. Whether nurses trained in adult nursing have the necessary skills, knowledge and attitudes to care for the parents is also discussed. Recommendations for ensuring care of the parent point to addressing skills gaps in adult-trained nurses and to reappraising visiting policies. PMID- 9873311 TI - Nurses' time management in intensive care. AB - During the 1980s, interest was shown in North America as to how Intensive Care Nurses use their work time, in response to a shortage of trained nurses. These studies were developed to investigate the amount of nursing time that could be saved by computerised recording systems. Similar pressures are now present in the United Kingdom, but there are no published work load studies of Intensive Care Nurses. This study used a five category tool to examine the work load of nurses in a cardiothoracic ICU. The methodology was designed so that comparison could be made with the earlier American studies. The 36 nurses studied spent 41% of their time in direct nursing care, 22% in patient assessment, 19% in clerical duties, 11% in time outside the unit and 7% in non-nursing duties. These findings were compared with the North American studies. Similarities were found which give some support to the reliability and validity of the tool. PMID- 9873312 TI - Developing a multidisciplinary protocol for enteral feeding. AB - The management of planned change is discussed using a change model. Protocols are described and defined. The process of multidisciplinary protocol development is identified. Nurses are encouraged to reflect upon the process of practice change. PMID- 9873313 TI - Positioning: one good turn after another? AB - Changing the position of a critically ill patient needs careful evaluation to prevent hypoxaemia. This article reviews the literature to determine how lateral and supine positioning can be used to maximise gas exchange in the critically ill. The relationship between reduced cardiac output and hypoxia is reviewed. Implications for nursing practice are discussed. PMID- 9873314 TI - Transcutaneous oxygen monitoring in congenital diaphragmatic abnormalities. AB - The article considers the use of transcutaneous (Tc) partial pressure of oxygen (PO2) monitoring as an alternative form of continuous monitoring of respiratory function in paediatric critical care. The principles of TcPO2 monitoring are discussed. The relationship between TcPO2, arterial blood gases and oxygen saturation is highlighted by examining trends in TcPO2 derived in three neonates with congenital diaphragmatic abnormality. The effect of critical incidents upon TcPO2 is also explored. TcPO2 is unlikely to enhance the assessment of ventilation provided by pulse oximetry and blood gas analysis. PMID- 9873315 TI - Reflections on being therapeutic and reflection. AB - This article offers a reflective account of an incident that occurred between a nurse tutor and a patient on a cardiology ward. It highlights the importance of interpersonal skills in creating a therapeutic relationship, in particular those of self-awareness, empathy and intuition. The author's difficulties in running reflective practice sessions for pre-registration students are discussed and insights are offered into why these difficulties arose. PMID- 9873316 TI - Nurses' under-medication of analgesia in cardiac surgical patients: a personal exploration. AB - This paper examines aspects of care which may account for some of the reasons why critical care nurses fail to relieve patients' pain following cardiothoracic surgery. Factors that may influence the critical care nurses' decision regarding the amount of opiate analgesia to give a patient are examined using the 'Theory of Planned Behaviour' as a framework for enquiry. The skills required by the critical care nurse in planning how to play the phenomena of 'the doctor-nurse game' may be a key element in meeting the goal of pain relief for the patient following cardiac surgery. PMID- 9873317 TI - Expert practice: what type of expert are you? PMID- 9873318 TI - Living on the edge--patients with an automatic internal cardioverter defibrillator (AICD): implications for nursing practice. AB - Automatic internal cardioverter defibrillator (AICD) therapy is an expanding treatment option for patients experiencing sudden cardiac death events [Author: is this the correct term? It sounds as if they have in fact died. Could we say 'experiencing sudden, potentially fatal, cardiac events'?]. The paper outlines the background to the technique and the nursing care of such patients and their families. It recommends that future nursing research should focus on exploring the impact, on the patients and their families, of living with this device. PMID- 9873319 TI - The holistic management of the patient with pre-eclampsia. AB - Physiological adaptations of pregnancy are summarised. Maternal consequences of pre-eclampsia, eclampsia and HELLP syndrome are described. Goals of ITU management are outlined. Directions for future management/therapies are included. The need to integrate technological expertise with sensitive nursing care is emphasised. PMID- 9873320 TI - The effects on nurses of treatment-withdrawal decisions made in ICUs and SCBUs. AB - This descriptive study explores the effects on nurses of treatment-withdrawal decisions made in ten ICUs and two SCBUs in Southern England. The study focused on the decision-making process, the reasons given in support of a decision to withdraw, the actions taken subsequently and the nurses' roles throughout the proceedings. Nurses' feelings were then explored about the decision, their ability to participate in ethical debate, the amount of influence they had on events and about the best and worst aspects of the situations described. PMID- 9873321 TI - Professional competence--can it be accurately measured or defined? AB - Professional competence is a complex and dynamic concept. The literature is explored and definitions of competence discussed. The paper identifies the current difficulties in defining and measuring competence. Recommendations are to identify the importance of self-assessment, clinical supervision and reflective practice as components of professional practice. PMID- 9873322 TI - Clinical decisions made by nurses in intensive care--results of a telephone survey. AB - A telephone survey of 89 GICUs in England was performed, using stratified quota sampling. 75 (84%) units reported changes in practice over the past 3 years. The reported changes were not only task orientated but also encompassed increased autonomy, organisational changes and increased workload. The absence of significant relationships between the activities suggests that nursing practice development is dependent on patient need and/or local factors. The lack of pattern suggests that it may be inappropriate to compare units for the purpose of nurse staffing. PMID- 9873323 TI - Turning a patient prone with ARDS. AB - This case report outlines the treatment and management of a patient who developed ARDS. The causes of ARDS and disordered physiology are discussed briefly. Prone positioning is identified as an important component of management. PMID- 9873324 TI - Creativity, innovation and expertise: reflections on BACCN conference 97. PMID- 9873325 TI - The experiences and health care needs of Asian coronary patients and their partners. Methodological issues and preliminary findings. AB - The incidence of heart disease in the Asian population in the UK is high and mortality from coronary heart disease for both Asian men and women is also higher than for non-Asians. The need to provide more information on cardiac rehabilitation programmes is highlighted. Cultural factors influence how Asian people perceive the heart, health and health care practices. A number of methodological issues are pertinent to conducting research involving ethnic groups, namely inter vs intra group differences, victim blaming, sampling, measurement, language issues and researcher effect. The aim of the study is to design, refine and validate a number of tools for use with Asian people in relation to cardiac rehabilitation. Preliminary and tentative conclusions suggest that cardiac rehabilitation services need a higher profile within the Asian community and that established approaches to data collection may need to be challenged if accurate and relevant feedback is to be achieved. PMID- 9873326 TI - The problems inherent in promoting research-based practice and strategies by which they might be overcome. AB - The historical and contemporary impetus for research in nursing is presented in brief. The factors affecting research use in practice are numerous and complicated and include political, financial, ideological and professional spheres. Various strategies that may enhance research use in practice have been suggested and evaluated and these are discussed. PMID- 9873327 TI - Critical care in a thinking environment: a personal perspective. AB - This paper is a transcript of a presentation made at the BACCN Conference. It highlights from the personal experience of the author a number of behaviours important in creating a thinking environment. Four areas are specifically focused on: giving colleagues attention; appreciating each other; management of meetings; and challenging limited assumptions. PMID- 9873328 TI - Technology-dependent children at home. AB - Many children cared for at home are dependent on medical technology. Stressors for the family caring for technology dependent children include: finances and adaptation to new roles. Good discharge planning is the key to successful adaptation by the family as discharge home is often delayed by financial wrangles between purchasers and providers. PMID- 9873329 TI - The impact of the ageing population on intensive care provision. AB - This paper explores the impact of the ageing population on the health service and intensive care provision. The concept of rationing is discussed. The paper concludes that age alone is not a reliable prediction of outcome (e.g. length of stay; mortality). The review highlights the lack of literature available offering a comparison of costs associated with intensive care management of the elderly. PMID- 9873330 TI - An unsafe environment: the reflection of a clinical manager. AB - This reflection considers of the actions of the author, a junior ward sister working on a cardiology ward. The reflection is triggered by the unsafe environment found when arriving on the ward unexpectedly, and the report from a staff member. It explores the actions taken in relation to the staff involved, considers the implications of being an advocate for both patients and staff in practice, and focuses on creating an environment where staff feel they can speak out. PMID- 9873331 TI - Inhaled nitric oxide: a possible therapy for ARDS. AB - Ten years ago' endothelium derived relaxing factor' was identified as nitric oxide (NO.). This highly significant discovery revealed the importance of NO. in normal physiology and pathophysiology. Research over the past decade into the potential therapeutic use of inhaled NO. in the management of ARDS is reviewed. In critical care, inhaled NO. seems to produce selective pulmonary vasodilation and this is already beginning to have an impact on the management of lung injuries including ARDS. The effect of NO. in multi-system failure is not yet established. Formal evaluation in the form of clinical trials has yet to be undertaken, and further study of all the potential side effects and toxicity are required for conclusive evidence of the value of inhaled NO. in the treatment of ARDS. PMID- 9873332 TI - Managing the nursing resource. PMID- 9873333 TI - Assessment of parental competence to consent to treatment for their child. AB - The admission of a child to the intensive care unit (ICU) represents a traumatic experience for parents. They may feel bombarded with information, loss of control and subject to enormous stress. The psychological stress of their child being admitted to ICU may render parents incapable of rational decision-making. Parents may receive frequent requests for consent at a time when their mental competence to do so may be questioned. Parental competence is affected by: information giving, effective communication and the perception of the person evaluating. The purpose of assessment of mental competence is to ensure that informed consent takes place and the best interests of the child are protected. PMID- 9873334 TI - Advanced practice: do we know what it is? AB - The UKCC has proposed the development of an advanced practitioner role. The role is equated with master's and doctorate level of preparation. The development of the advanced practitioner role is outlined. The assumption that the nursing profession understands the term advanced practice is challenged. It is not possible to make explicit the role of the advanced practitioner unless nursing knowledge is used as its foundation. Two basic questions have yet to be answered: What is advanced practice? What is an advanced practitioner? PMID- 9873335 TI - Contracting for intensive care services. AB - Purchasers will increasingly expect clinical services in the NHS internal market to provide objective measures of their benefits and cost effectiveness in order to maintain or develop current funding levels. There is limited scientific evidence to demonstrate the clinical effectiveness of intensive care services in terms of mortality/morbidity. Intensive care is a high-cost service and studies of cost-effectiveness need to take account of case-mix variations, differences in admission and discharge policies, and other differences between units. Decisions over development or rationalisation of intensive care services should be based on proper outcome studies of well defined patient groups. The purchasing function itself requires development in order to support effective contracting. PMID- 9873336 TI - Intensive care nurses' perceptions of stress. AB - The study aimed to provide a description of intensive care nurses' perceptions of stress, frequency and demand (tension and tiredness) in relation to potentially stressful events encountered at work. The study sought to examine the relationship between perceived levels of stress, frequency and demand. Caution needs to be applied in the interpretation of the findings, due to the limitations of the research design. The findings appeared to suggest that a number of events are perceived as stressful and make a demand on individual nurses, irrespective of the frequency of occurrence or the level of knowledge and expertise of the nurse. The findings also supported the notion that perceived degree of control, available resources and appropriate clinical support are instrumental in enabling nurses to cope with stress and the demands generated by intensive care nursing. PMID- 9873337 TI - Exploring the psychological effects of intensive care on paediatric patients: issues from the literature. AB - The psychological needs of children in intensive care may often be neglected in favour of a more physiological approach to nursing care. Both the short- and long term psychological effects of paediatric intensive care require examination. There is a lack of research-based evidence on the psychological effects of intensive care on children. The intensive therapy unit (ITU) itself, developmental age, role of the family, loss of self-control/routine, under/over use of sedation and analgesia, play, and the ITU nursing staff all have a significant psychological impact on the critically ill child. Strategies that reduce the negative psychological effects of ITU must address the above issues. PMID- 9873338 TI - Job satisfaction of intensive care nurses practising primary nursing. A comparison with those practising total patient care. AB - This study compares the job satisfaction experienced by intensive care nurses practising primary nursing with those who practised total patient care in an a earlier study using the same tool. Differences in the job aspects of; challenge, initiative, seeing work through, output/goals, knowledge and skills appear to be associated with the organisational approach. Some similarities exist between the two units-nurses gained most sense of achievement from: seeing patients improve, personal achievement, giving care and camaraderie. In both units nurses wanted greater opportunities for inter-professional involvement and decision-making. Primary nursing as an organisational approach may provide opportunities for increasing the amount of challenge, decision-making, opportunities for using initiative, seeing work through and subsequently job satisfaction in intensive care nursing. Further study of the job satisfaction of nurses practising primary nursing in ICU is recommended, but sizes and return rates need to be optimised. PMID- 9873339 TI - Reflection on a patient receiving high frequency oscillatory therapy. AB - This reflection on practice is written by two members of a primary nurse team at the Chelsea and Westminster intensive care unit (ICU)/nursing development unit (NDU). Johns's (1993) tool for structured guided reflection was used to reflect on the care of an ICU patient given by her primary and associate nurse. The issues highlighted include: The importance of knowing your patient through a primary nursing approach. Collaboration between the primary nursing team and the anaesthetists. Negotiation between the patient, her family and the primary nursing team. Empowerment of the patient to express her view. PMID- 9873340 TI - For the record. PMID- 9873341 TI - Changing nurses' practice. PMID- 9873342 TI - Am I dying, nurse? PMID- 9873343 TI - The effect of care planning on quality of patient care. AB - Nursing care plans can be seen as a means of ensuring holistic care. This study investigated the view of nurses and support workers on the care plan as a tool to enhance quality care and how their views were reflected in practice. PMID- 9873344 TI - Exploring new roles for nurses in the acute sector. AB - Recent changes in the NHS have resulted in numerous developments in nursing roles. A project has been undertaken to identify the wide range and purpose of extended nursing roles, and to discover the differences between them. PMID- 9873345 TI - The management of acute pain in children. AB - Children cannot always articulate the level of pain they are experiencing, and need to be assessed carefully. The final article in this three-part series discusses a number of pain assessment tools available to help nurses caring for children in pain. PMID- 9873346 TI - Hand-held Dopplers in central catheter insertion. AB - Peripherally inserted central catheters benefit many patients requiring repeated intravenous treatment. This study investigated the effect of using a hand-held Doppler to aid insertion where a patient's veins are not visible or palpable. PMID- 9873347 TI - Ileostomy. AB - Several different groups of patients may undergo an ileostomy, the two most common being those with ulcerative colitis and and Crohn's disease. Nurses have an important role in providing information, advice and support to such patients. PMID- 9873348 TI - Atopic eczema and wet-wrap dressings. AB - When atopic eczema does not respond to first-line treatments, wet-wrap dressings may provide a solution. Once the technique has been mastered, patients may find it gives them control over their condition. PMID- 9873349 TI - Implementation of change: a wound assessment chart. AB - The implementation of change involves careful planning, particularly if it depends on the participation of others, such as the introduction of a new nursing tool. Those involved must be educated about the change so they can influence developments. PMID- 9873350 TI - Equipment for moving and handling patients. AB - Choosing the correct equipment for moving and handling patients is essential to prevent injuries among health-care staff and patients' carers. It is vital, however, that a thorough needs and risk assessment is undertaken before equipment is purchased. PMID- 9873351 TI - Bereavement Part 2: Breaking bad news. AB - Breaking bad news, whether telling a patient about a poor prognosis or informing relatives of the sudden death of a loved one, is one of the most daunting tasks facing nurses. It requires a sensitive approach, based on individuals' right to know the truth. PMID- 9873352 TI - Nocturnal enuresis. AB - Nocturnal enuresis (NE), the involuntary passing of urine during sleep after the age at which bladder control would normally be anticipated, is a widespread and potentially disabling disorder for children. The treatment of NE constitutes several approaches and its pathophysiology remains unsolved. Careful consideration should be given to the work-up of NE since there may be concurrent symptoms that require attention either before or in conjunction with the treatment. Patient/family education and a cooperative approach usually produce the most favorable results in treating NE. PMID- 9873353 TI - Impotence and perceived partner support. AB - Because impotence has both physiologic and psychologic components, it seems that impotent patients might benefit from a holistic approach to treatment that includes a medical regimen for physiologic factors as well as treatment for social and psychologic factors. The spousal relationship is one social and/or psychological factor. Often, a male experiences his sexual dysfunction as profoundly stressful while the partner finds it annoying or disturbing, but not threatening to the overall bond between them (Rust, Golombok, & Collier, 1988). This study was conducted to explore patient perception of the spousal (partner) relationship during admission to an erectile dysfunction clinic. PMID- 9873354 TI - A retrospective review of outcomes in one clinic's treatment of urinary incontinence. AB - Two options usually presented to patients for managing and treating chronic incontinence are surgery or medications. Lack of third-party payment for other noninvasive treatment options led to the development of a center to treat people with chronic incontinence/frequency who might not desire or need costly work-up by a urologist or gynecologist. It was hoped that by demonstrating the cost effectiveness of ancillary providers, procedures, and equipment, third-party payers would be more generous in their coverage. PMID- 9873355 TI - The culture of long-term care: impact on a continence care program. AB - Continence care in long-term care settings is a major caregiver and economic burden, with incontinence affecting an estimated 50% to 70% of all residents. Traditionally the continence care of residents is assigned to the nursing assistants and continence programs have been on paper only. These programs have had dismal success rates. Factoring in the culture of the long-term care setting can make the continence program more comprehensive and improve success rates. PMID- 9873356 TI - Pelvic muscle rehabilitation in males following prostatectomy. AB - Post-prostatectomy incontinence (PPI) is a common problem in the era of increased detection of prostate cancer and use of radical prostatectomy. Whether temporary or permanent, mild or more severe, PPI can be effectively treated and improved by pelvic muscle rehabilitation. It is important for urologic nurses to understand the various pelvic muscle rehabilitation methods--for example, Kegel exercises, biofeedback, and electrical stimulation--to better educate and care for PPI patients and their families. PMID- 9873357 TI - Evangelists needed. PMID- 9873358 TI - Getting ready for certification: male reproduction. PMID- 9873359 TI - Getting ready for advanced practice certification: urolithiasis, Part II. PMID- 9873360 TI - Psychometric evaluation of the international prostate symptom score. AB - The International Prostate Symptom Score represents an important development in the understanding of benign prostatic hyperplasia (BPH) and its clinical management. This instrument provides reasonable reliability, content validity, and predictive validity. However, it is limited by a lack of concurrent validity so it can not be used to discriminate the voiding dysfunction associated with BPH from that associated with unrelated conditions. Psychometric properties of this symptom score are evaluated and suggestions provided for increased involvement for urologic nurses in its clinical application and future development. PMID- 9873361 TI - Self-esteem and depression in men who present with erectile dysfunction. AB - Erectile dysfunction always has a psychologic component in addition to the underlying physical cause. The extent of depression and reduced self-esteem in patients who present with erectile dysfunction are explored in this study. Suggestions are given for how urologic nurses can overcome patients' fears and concerns. PMID- 9873362 TI - Improved patient outcomes at a urology nurse clinic. AB - A urology nurse clinic was developed to improve cost and quality of care outcomes for patients with benign prostatic hypertrophy amid cost constraints and the need for improved service delivery. Cost and quality of care outcomes were achieved through earlier discharge and improved assessment, education, and discharge planning for patients. The professional development of urology nurses also improved through increased practice of advanced assessment skills and broadened scope of practice. PMID- 9873363 TI - Prostate disease patients: planning services to meet their coping needs. AB - A committee comprising hospital and community-based urology and oncology nurses and social workers planned, organized, implemented, and evaluated an educational public seminar on prostate cancer (PC). Data relating to satisfaction with the seminar, reasons for attendance, perceived needs for further support/education, and demographics were collected using a feedback questionnaire. Results suggested a need for education and a significant interest in support groups and further educational forums for patients with PC or symptoms and their caregivers. PMID- 9873364 TI - Medication use by community-dwelling elderly with urinary incontinence. AB - Medication consumption patterns among community-dwelling elderly persons with chronic urinary incontinence were examined. All health professionals, including nurses, should be knowledgeable of the potential for adverse drug reactions. Continuing education and establishment of routine monitoring systems could help to reduce the risk of potential for adverse drug reactions and prevent drug induced UI in patients. PMID- 9873365 TI - Getting ready for certification: voiding dysfunction. PMID- 9873366 TI - Fluoroquinolone antibiotics. PMID- 9873367 TI - Broad spectrum matrix metalloproteinase inhibitors: an examination of succinamide hydroxamate inhibitors with P1 C alpha gem-disubstitution. AB - A series of P1 C alpha gem-disubstituted succinamide hydroxamate matrix metalloproteinase inhibitors were prepared stereoselectively and evaluated in vitro for their ability to inhibit MMP-1, MMP-2, and MMP-3. It was found that while methyl/allyl substitution as in 2 and 18 provided compounds that were broad spectrum inhibitors and nearly equipotent with parent inhibitor 1, a larger group such as bis-allyl as in 13 or gem-cyclopentyl as in 14 significantly reduced enzyme inhibition. PMID- 9873368 TI - Relationship between dihedral angles of N1 and C9 substituents in 1,4 benzodiazepines and dual cholecystokinin-A and -B antagonistic activities. AB - Introduction of a methyl moiety to the C9 position of a 1,4-benzodiazepine ring system afforded dual CCK-A and -B antagonistic activity. Novel derivatives having ethyl, isopropyl and chloro substituents at C9 were prepared in order to obtain more potent antagonistic activities. AM1(MOPAC93) calculations of the dihedral angles between the N1 and C9 substituents indicated that dihedral angles for dual antagonistic activities were between 50 degrees and 60 degrees. A methyl moiety was selected as the most suitable C9 substituent in this series for potent dual CCK-A and -B receptor antagonistic properties. PMID- 9873369 TI - Functionalization of a viscosity-sensitive fluorophore for probing of biological systems. AB - Functionalization of a viscosity-sensitive visible wavelength fluorophore 2-(1,1 dicyanopropenyl-2)-6-dimethylaminonaphthalene (DDNP), with the intent to incorporate its favorable optical properties into a probe for structural and functional imaging by fluorescence microscopy, is described. Spiperone, a highly potent ligand for the dopamine D2 receptors, was conjugated via an ethylpiperazine moiety to the fluorophore giving fluorescent probes that can be excited in the UV and Vis range. PMID- 9873371 TI - Inhibition of electric eel acetylcholinesterase by porphin compounds. AB - Synthetic porphin compounds have been found to be reversible inhibitors of acetylcholinesterase from electric eel with Ki values of microM range. It seems that the number and position of fluorine on the phenyl ring and metal of an inhibitor play an important role for binding of an inhibitor to the enzyme active site. PMID- 9873370 TI - 5-Alkyl-2-[(methylthiomethyl)thio]-6-(benzyl)-pyrimidin-4-(1H)-ones as potent non nucleoside reverse transcriptase inhibitors of S-DABO series. AB - Novel dihydroalkoxybenzyloxopyrimidine (S-DABO) derivatives targeting the non nucleoside inhibitor (NNI) binding site of human immunodeficiency virus (HIV) reverse transcriptase (RT) have been synthesized using a novel computer model for the NNI binding pocket and tested for their RT inhibitory activity in cell-free assays using purified recombinant HIV RT as well as for their anti-HIV activity in HTL VIIIB-infected peripheral blood mononuclear cells. Our computational approach allowed the identification of several ligand derivatization sites for the generation of more potent S-DABO derivatives. Our lead S-DABO derivative, 5 isopropyl-2-[(methylthiomethyl)thio]-6-(benzyl)-pyrimidin-4-(1H)-one (compound 3), elicited potent anti-HIV activity with an IC50 value of less than 1nM for inhibition of HIV replication without any evidence of cytotoxicity and an unprecedented selectivity index of > 100,000. PMID- 9873372 TI - 2(1H)-quinolinone derivatives as novel anti-arteriostenotic agents showing anti thrombotic and anti-hyperplastic activities. AB - In order to search for anti-arteriostenotic agents, a series of 2(1H)-quinolinone derivatives was synthesized and evaluated for anti-thrombotic activity and for anti-hyperplastic activity. From this series, (-)-6-[3-[3-cyclopropyl-3-[(1R,2R) 2-hydroxycyclohexyl]ureido]propoxy]-2 (1H)-quinolinone (1p, OPC-33509) was selected as the best candidate by balancing the efficacy on anti-thrombosis and anti-hyperplasia. PMID- 9873373 TI - Peptidyl beta-homo-aspartals: specific inhibitors of interleukin-1 beta converting enzyme and its homologues (caspases). AB - Inhibition of interleukin-1 beta converting enzyme (ICE), apopain, papain, thrombin and trypsin with substrate like peptidyl L- and D-alpha-aldehydes and their L-beta-homo-aldehyde analogues was investigated. The L-beta-homo-aspartals appear to be specific inhibitors for ICE and its homologues; the other enzymes were not inhibited with such L-beta-homo aldehydes. Papain shows tolerance for D residues at P1 depending on their chiral stability. PMID- 9873374 TI - Anti-AIDS agents. 31. Synthesis and anti-HIV activity of 4-substituted 3',4'-di-O (-)-camphanoyl-(+)-cis-khellactone (DCK) thiolactone analogs. AB - Four DCK-thiolactone analogs (3-6) were synthesized asymmetrically and evaluated for anti-HIV activity against HIV-1 replication in H9 lymphocyte cells. Based on the functionality on the thiolactonecoumarin nucleus, activity was in the order: methyl > H > propyl > benzyl. 4-Methyl-3',4'-di-O-(-)-camphanoyl-(+)-cis khelthiolactone (4) exhibited extremely potent anti-HIV activity with EC50 and therapeutic index values of 0.00718 microM and > 21,300, respectively. PMID- 9873375 TI - 8-Aminoquinolines as anticoccidials--II. AB - During a chemistry program aimed at finding a novel analogue of pentaquine with improved in vivo activity, a number of hypotheses concerning the way this drug acts in the chicken were investigated. Consideration of the products of monoamine oxidase metabolism of pentaquine suggested that pentaquine aldehyde is the likely active metabolite. Although isolation of this unstable compound was not possible, oxime and cyclic acetal and ketal derivatives were obtained and shown to possess in vitro anticoccidial activity. PMID- 9873376 TI - N-[4-(1,1'-biphenyl)methyl]-4-(4-thiomorpholinylmethyl) benzenamines as non oxazolidinone analogues of antimycobacterial U-100480. AB - Thiomorpholine analogues of U-100480 with the biphenylmethyl group replacing the acetamidomethyloxazolidinone moiety have been synthesized and tested as antimycobacterial agents together with various related derivatives. Some biphenyl derivatives were endowed with high activity against Mycobacterium tuberculosis and other non-tuberculous mycobacteria. PMID- 9873377 TI - Isoindolinone enantiomers having affinity for the dopamine D4 receptor. AB - PD 108635 (1) was identified as a potent dopamine D4 ligand and we wanted to replace the benzylic alcohol with a metabolically more stable moiety. Investigations led to the discovery of a series of isoindolinones having D4 affinity. PMID- 9873378 TI - Syntheses of two enantiomeric pairs of myo-inositol(1,2,4,5,6) and -(1,2,3,4,5) pentakisphosphate. AB - Two enantiomeric pairs of myo-inositol(1,2,4,5,6)P5 and -(1,2,3,4,5)P5 have efficiently been synthesized by means of the lipase catalyzed acetylation of 1,2:5,6-di-O-isopropylidene-myo-inositol and the benzoyl migration procedure. PMID- 9873379 TI - Synthesis and pharmacological activities of 13-dehydro derivatives of primary prostaglandins. AB - 13-Dehydro derivatives of prostaglandin E1, E2, E3, F1 alpha and F2 alpha were synthesized. Compared with natural prostaglandins, 13-dehydro analogues were found to exhibit more potent inhibitory activity against human platelet aggregation and relaxation of guinea-pig isolated trachea, while they showed less potent activity of contraction of guinea-pig isolated ileum. PMID- 9873381 TI - Synthesis of protected guanidinium linked dinucleoside incorporable into an oligonucleotide using solid phase DNA methodology. AB - The synthesis of novel fully protected guanidinium linked dinucleoside for incorporation into oligonucleotide using solid-phase DNA synthesis methodology was developed. The three different protecting groups selected allow different deprotection conditions. PMID- 9873380 TI - Synthesis and anti-HIV activities of urea-PETT analogs belonging to a new class of potent non-nucleoside HIV-1 reverse transcriptase inhibitors. AB - A series of potent specific HIV-1 RT inhibitory compounds is described. The compounds are urea analogs of PETT (PhenylEthylThiazoleThiourea) derivatives and the series includes derivatives with an ethyl linker (1-6) and conformationally restricted analogs (7-13). The antiviral activity is determined both at the RT level and in cell culture on both native and mutant forms of HIV-1. Many compounds display activity in the nM range against wt-RT. PMID- 9873382 TI - Evaluation of phosphorus-containing inhibitors of gamma-glutamyl hydrolase. AB - Several putative, phosphorus-containing inhibitors of gamma-glutamyl hydrolase were synthesized and evaluated for inhibitory activity. The phosphonamidoic acids were shown to be weak competitive inhibitors while both a phosphoramidate diester and a phosphonamidate ester were shown to be potent time-dependent inactivators, presumably through irreversible phosphorylation of an active site nucleophile. PMID- 9873383 TI - Synthesis and in vitro cytotoxicity of lipophilic platinum(II) complexes. AB - A number of lipophilic platinum(II) complexes of the general structures cis [Pt(LA)2Cl2] and [Pt(LD)Cl2] were synthesised. Long chain amines (LA) and diamines (LD), prepared from lipidic amino acids, were used as ligands. The in vitro cytotoxicity of the complexes was evaluated against four cell lines (P388, NSCLC-N6, E39, M96). cis-Dichloro-bis(2-aminohexadecanol)platinum(II) was the most active against P388, NSCLC-N6 and E39 (IC50: 11 micrograms/ml, 25 micrograms/ml, 31 micrograms/ml), while dichloro(1,3 heptadecanediamine)platinum(II) presented the highest activity against M96 (IC50: 13 micrograms/ml). PMID- 9873384 TI - Design and synthesis of an orally active GPIIb/IIIa antagonist based on a phenylpiperazine scaffold. AB - The design and synthesis of an orally active LMW non-peptide GPIIb/IIIa antagonist, based on a N,N'-bisphenylpiperazine scaffold, is described. The optimal compound showed a high in vitro binding potency (pIC50 = 8.7) in combination with potent oral antithrombotic activity (30-40% inhibition of thrombus growth at 0.3-3 mg/kg) with a duration of action of > 90 min. in a hamster cheek pouch model. PMID- 9873385 TI - A potent dipeptide inhibitor of dipeptidyl peptidase IV. AB - A series of novel potent inhibitors of dipeptidyl peptidase IV (DPP-IV) has been developed. A brief structure-activity relationship of the inhibitors was investigated. The dipeptide TSL-225, tryptophyl-1,2,3,4-tetrahydroisoquinoline-3 carboxylic acid, was identified with the critical structure for the inhibitory activity. PMID- 9873386 TI - Synthesis and NK1 receptor antagonistic activity of (+/-)-1-acyl-3-(3,4- dichlorophenyl)-3-[2-(spiro-substituted piperidin-1'-yl)ethyl]piperidines. AB - (+/-)-1-Acyl-3-(3,4-dichlorophenyl)-3-[2-(spiro-substituted piperidin-1' yl)ethyl]piperidines and their quaternary ammonium salts were prepared and evaluated for their NK1 receptor antagonistic activity. Some of these inhibited SP-induced contraction in guinea pig ileum with IC50 values at a level of 10(-9) M and showed potent inhibitory activity against selective NK1 receptor agonist induced bronchoconstriction in guinea pigs. PMID- 9873388 TI - Synthesis of N-(1-ethyl-4-methylhexahydro-1,4-diazepin-6-yl)nicotinamides and their affinities for 5-HT3 and dopamine D2 receptors. AB - A series of N-(1-ethyl-4-methylhexahydro-1,4-diazepin-6-yl)nicotinamide derivatives were prepared and evaluated for their binding to 5-HT3 and dopamine D2 receptors. Among them, the 5-bromo-2-methoxy-6-methylaminonicotinamide 16 and its (R)-isomer were found to have potent affinities for both receptors. The affinities of (R)-16 for 5-HT3 and dopamine D2 receptors are approximately 3-fold higher than those of the corresponding benzamide (R)-1 (IC50: 1.1 and 12 nM vs. 2.9 and 35 nM, respectively). PMID- 9873387 TI - Synthesis and antitumor activity of 4-phenyl-1-arylsulfonyl imidazolidinones. AB - Novel 1-(1-benzoylindoline-5-sulfonyl)-4-phenyl-4,5-dihydroimidazolon es 3 synthesized show highly potent and broad cytotoxicities. Among them compound 3b (DW2143) exhibits much more potent cytotoxicities than doxorubicin and highly effective antitumor activities against murine (3LL, Colon 26) and human xenograft (NCI-H23, SW620) tumor models. PMID- 9873389 TI - Modulation of multidrug resistance by taxuspine C and other taxoids from Japanese yew. AB - Taxuspine C (1), a new taxoid from the Japanese yew Taxus cuspidata, increasing the cellular accumulation of vincristine (VCR) in multidrug-resistant tumor cells as potent as verapamil enhanced the chemotherapeutic effect of VCR in P388/VCR bearing mice. When taxuspine C (1) was given i.p. daily at 200 mg/kg with 0.2 mg/kg VCR for 5 days, a treated/control (T/C) value of 138% was obtained. The other new taxoids, taxezopidines G (8) and H (9), from the yew also increased the VCR accumulation as potent as verapamil. These results suggest that some taxoids may be useful for overcoming multidrug resistance in tumor cells. PMID- 9873390 TI - A new class of sulfoxide surfactants derived from Tris. Synthesis and preliminary assessments of their properties. AB - A new class of non-ionic amphiphilic molecules suitable for biological purposes, especially extraction of membrane proteins, is reported. Such surfactants were prepared in two steps: addition of alkyl or fluoroalkyl mercaptan on Tris(hydroxymethyl)acrylamidomethane (THAM) derivatives, followed by the oxydation of sulfide group in sulfoxide moiety in order to provide water solubility to the molecule. The detergent efficiency of these new surfactants were assayed on rat liver cells. PMID- 9873391 TI - Synthesis and pharmacology of N-alkylated derivatives of the excitotoxin ibotenic acid. AB - Three amino-alkylated derivatives of the naturally occurring excitatory amino acid (EAA) receptor agonist ibotenic acid (Ibo) have been synthesized and tested pharmacologically. N-Methyl-Ibo (1a) and N-ethyl-Ibo (1b) were shown to be agonists at NMDA receptors (EC50 = 140 and 320 microM, respectively), though with activities considerably lower than Ibo (EC50 = 9.6 microM). N-Benzyl-Ibo (1c) was inactive at ionotropic EAA receptors and all three compounds were, in contrast to Ibo, inactive at metabotropic EAA receptors. Molecular mechanics calculations have been performed on Ibo, 1a-c and the potent NMDA agonist 2-amino-2-(3-hydroxy 5-methyl-4-isoxazolyl)acetic acid (AMAA) in order to elucidate the observed structure-activity data. PMID- 9873392 TI - Synthesis and preliminary evaluation of (S)-2-(4'-carboxycubyl)glycine, a new selective mGluR1 antagonist. AB - The synthesis and the pharmacological evaluation at metabotropic glutamate receptors of S-2-(4'-carboxy-cubyl)glycine (ACUDA, 9) are described. S-2-(4' Carboxy-cubyl)glycine is a structurally novel group I selective antagonist for mGluR1 subtype. PMID- 9873393 TI - On the preparative use of recombinant pig alpha(1-3)galactosyl-transferase. AB - A series of non-natural N-acyl derivatives of lactosamine is incubated with recombinant alpha(1-3)galactosyl-transferase and UDP-galactose. The enzyme shows a high promiscuity towards the non-natural acceptors. It selectively transfers a galactose unit onto the 3-OH group of the terminal beta-linked galactose in an alpha-mode to give an array of linear-B trisaccharides. PMID- 9873394 TI - Novel 1,3-disubstituted-5,10-dihydro-5,10-dioxo-1H-benzo[g] isochromene-3 carboxamides as potent antitumor agents. AB - Novel antitumor 5,10-dihydro-5,10-dioxo-1H- benzo[g]isochromene-3-carboxamides were discovered. PMID- 9873395 TI - Asymmetric synthesis and anti-HIV activity of L-carbocyclic 2',3'-didehydro-2',3' dideoxyadenosine. AB - Asymmetric synthesis of L-carbocyclic 2',3'-didehydro-2',3'-dideoxyadenosine and its analogs were accomplished and their anti-HIV activities were evaluated. It was found that L-carbocyclic 2',3'-didehydro-2',3'-dideoxyadenosine exhibited moderately potent anti-HIV (EC50 = 2.4 microM) activity in human PBM cells without cytotoxicity up to 100 microM. PMID- 9873397 TI - In vitro evaluation and crystallographic analysis of a new class of selective, non-amide-based thrombin inhibitors. AB - We describe the in vitro evaluation and crystallographic analysis of a new class of potent and selective, non-amino acid-based, small-molecule thrombin inhibitors, exemplified by 14. This class of achiral inhibitors lacks an amide based backbone, exhibits nM inhibition of thrombin, and is selective for thrombin. Compound 14 does not interact with the active-site catalytic apparatus and is anchored to the enzyme via a single network of hydrogen bonds to Asp189 of the S1 pocket. PMID- 9873396 TI - Synthesis and biological evaluation of a series of 2'-fluorinated-2',3'-dideoxy 2',3'-didehydro-(L)-nucleosides. PMID- 9873398 TI - Novel modified tripeptide inhibitors of alpha 4 beta 7 mediated lymphoid cell adhesion to MAdCAM-1. AB - MAdCAM-1 specifically binds the lymphocyte integrin alpha 4 beta 7 and participates in the homing of leukocytes to intestinal mucosal sites. The LDT sequence located on the CD loop of MAdCAM-1 is an important binding site for MAdCAM-1/alpha 4 beta 7 interactions. N-Terminus acylation of the LDT motif and modification of the C-terminus carboxamide with amines led to low micromolar MAdCAM-1 inhibitors. PMID- 9873400 TI - Diarylsulfonamides as selective, non-peptidic thrombin inhibitors. AB - Based on the structures of aminopyridine thrombin inhibitors (1), a series of aminoalkyl- and guanidinoalkyl-substituted diarylsulfonamides were prepared. The most potent derivative, N-[3-(4-guanidinobutoxy)-5-methyl-phenyl] benzenesulfonamide (6c) had Ki = 0.18 microM for thrombin and did not inhibit trypsin, plasmin, or factor Xa. Comparison of the X-ray structures of the thrombin/1b and the thrombin/6c complexes revealed important aspects which govern the binding of such diarylsulfonamides to thrombin. PMID- 9873399 TI - Synthesis and biological properties of new 1 beta-methylcarbapenems. AB - The synthesis and biological activity of the novel series of 1 beta methylcarbapenems, 1 and 2 were described. Most compounds displayed high potent antibacterial activity. The best compound in this series, 2a (IH201; R2 = NH2) showed an excellent and a broad spectrum as well as high renal DHP-I stability. It also possessed good in vivo efficacy and high safety. PMID- 9873401 TI - Practical synthesis of Taxol side chain. AB - Practical large scale synthesis of N-benzoyl-(2R,3S)-phenylisoserine methyl ester of the Taxol side chain has been attained from the coupling of chiral imine of N [(S)-methylbenzyl]benzaldimine with (Z)-alpha-methoxy trimethylsilyl ketene acetal followed by the sequential reactions of lactamization, demethylation, methanolysis and N-benzoylation. PMID- 9873402 TI - Carbohydrate modifications in the spirostane cellobioside cholesterol absorption inhibitor series. AB - Cholesterol absorption inhibition remains an attractive approach for the treatment of hypercholesterolemia. We have continued our SAR development in the spirostanyl cellobioside class of agents seeking a greater understanding of the role carbamoyl substitution has on the potency in this series. In this regard, a series of differentially substituted carbamate analogs were made with and without deoxygenations. From this study, it was determined that the minimal requirements for optimal potency was a lone carbamate at C4" and deoxygenation at the C6" position. PMID- 9873403 TI - A new series of cyclic amino acids as inhibitors of S-adenosyl L-methionine synthetase. AB - Optically active 3-amino-3-(tetrahydrofuranyl) carboxylic acid, 3-amino-3 (tetrahydrothienyl) carboxylic acid and their corresponding six membered ring analogues have been synthesised and examined as potential inhibitors of the enzyme S-adenosylmethionine (AdoMet) synthetase. The kinetic behaviour of these compounds was studied using recombinant rat liver AdoMet synthetase (alpha isoform) fractionated from E. coli transformed with the plasmid pSSRL-T7N. All the compounds tested were competitive inhibitors of the enzyme with respect to L methionine. PMID- 9873404 TI - Synthesis and biological activity of lactones en route to Altohyrtin A. AB - Lactones 2 and 7 were synthesised and tested against six human tumour cell lines (Pancreas-a, BXPC-3), (Thyroid ca, KAT-4), (Thyroid ca, SW1736), (Lung-NSC, NCI H460), (Pharynx-sq, FADU) and (Prostate, DU-145). Lactone 7 proved inactive, but lactone 2 displayed some activity against four of the six cell lines examined. Both lactones were converted into an intermediate 5 en route to Altohyrtin A. PMID- 9873405 TI - Adenosine deaminase inhibitors. Synthesis and biological evaluation of aralkyladenines (ARADS). AB - Several 9-aralkyladenines have been prepared and their ADA inhibitory activity was determined. A minimum of two carbon atoms separating the aromatic ring from the adenine-bearing carbon (C3') was found essential for potent activity. PMID- 9873406 TI - Inhibitors of the bacterial cell wall biosynthesis enzyme MurD. AB - A series of transition-state analog inhibitors of the D-glutamic acid-adding enzyme (MurD) of bacterial peptidoglycan biosynthesis has been synthesized and evaluated for inhibition of the E. coli enzyme. PMID- 9873407 TI - Thrombin receptor (PAR-1) antagonists. Heterocycle-based peptidomimetics of the SFLLR agonist motif. AB - The thrombin receptor (PAR-1) is activated by alpha-thrombin to stimulate various cell types, including platelets, through the tethered-ligand sequence SFLLRN. A series of azole-based carboxamides, designed after SFLLR, were synthesized and evaluated in vitro. The compounds inhibited platelet aggregation induced by SFLLRN-NH2 or alpha-thrombin, and blocked the binding of [3H]-S-(p-F-Phe)-Har-L Har-KY-NH2 to a CHRF membrane preparation of PAR-1. Oxazole 30 bound to PAR-1 with an IC50 of 1.6 microM, and gave IC50 values of 25 microM and 6.6 microM against alpha-thrombin- and SFLLRN-NH2-induced platelet aggregation, respectively. PMID- 9873408 TI - Synthesis and proteinase inhibitory properties of diphenyl phosphonate analogues of aspartic and glutamic acids. AB - The synthesis of diphenyl phosphonate analogues of aspartic and glutamic acid, and their inhibitory activity against S. aureus V8 protease and granzyme B, is described. The study has revealed difficulties with protecting group compatibility in the synthesis of these analogues. Two analogues, Acetyl. AspP (OPh)2 and Acetyl.GluP (OPh)2 were found to function as irreversible inactivators of V8 proteinase, yet exhibit no activity against granzyme B. PMID- 9873409 TI - Synthesis and acetylcholinesterase inhibitory activity of (+/-)-14 fluorohuperzine A. AB - The synthesis of (+/-)-14-Fluorohuperzine A has been accomplished and the ability of this agent to inhibit acetylcholinesterase has been measured. Taking into account its racemic form, this compound exhibits 62 times less potent activity than natural (-)-huperzine A. PMID- 9873410 TI - Solution and solid phase combinatorial synthesis of peptidomimetic library containing diversified alpha-methylated amino acids. AB - A combinatorial peptidomimetic library containing diversified alpha-methylated amino acids was generated by the Ugi four component condensation (4cc) reaction from acids, amines, isocyanides and ketones in both solution and solid phase synthetic procedures. This one-pot methodology overall gave fair to good yields, which compare well with multi-step syntheses. PMID- 9873411 TI - Identification of a potent analogue of Nazumamide A through iteration of combinatorial tetrapeptide libraries. AB - Five sets of N-acylated tetrapeptide libraries and sublibraries related to Nazumamide A have been prepared using 25 natural and unnatural amino acids. They were evaluated in antithrombin assay, in order to quantify inhibition at each step of the tetrapeptide sublibrary iteration. The studies led to the identification of 2,5-dihydroxybenzoyl-lysyl-isoleucyl-phenylalanyl-arginine as a novel inhibitor of thrombin and was found to be at least 25 times more potent than the natural tetrapeptide 2,5-dihydroxybenzoyl-arginyl-prolyl-isoleucyl-alpha aminobutyric acid (NAZA). PMID- 9873412 TI - N3-methyl-mafosfamide as a chemically stable, alternative prodrug of mafosfamide. AB - The presence of an alkyl substituent at N3 in the oxazaphosphorine ring stabilizes N-substituted 4-(alkylthio)cyclophosphamides from spontaneous decomposition. Based on this finding, N3-methyl-mafosfamide was synthesized and examined as a chemically stable, biooxidative prodrug of mafosfamide. This prodrug was stable in aqueous buffer (pH 7.4, 37 degrees C) and underwent N demethylation in a time dependent manner when incubated with rat hepatic microsomes. N3-Methyl-mafosfamide was 10-fold more cytotoxic in vitro than cyclophosphamide against mouse embryo Balb/c 3T3 cells (LC50 = 3.6 microM). Preliminary in vivo antitumor evaluation against L1210 leukemia in mice showed that this prodrug was active [Increase of life span (ILS) > 29%]. PMID- 9873413 TI - Conformational probes for elucidating the nature of substance P binding to the NK1 receptor: initial efforts to map the Phe7-Phe8 region. AB - Three substance P analogs with conformation constraints in the Phe7-Phe8 region have been prepared in connection with an effort to differentiate two families of potential conformations for the binding of substance P to its NK1 receptor. While the analogs did not bind the NK1 receptor with high affinity, the synthesis of the analogs demonstrated the utility of a general method for constructing piperazinone based peptidomimetics. PMID- 9873414 TI - Potent and efficacious thienylamidine-incorporated thrombin inhibitors. AB - Novel thrombin inhibitors incorporating thienylamidine at the P1 position were designed and synthesized. These compounds are potent, trypsin-selective and efficacious in the rat model of venous thrombosis. The proposed P1 binding mode in the thrombin active site was confirmed by X-ray crystallographic analysis. PMID- 9873415 TI - Carbaxylosides of 4-ethyl-2-oxo-2H-benzopyran-7-yl as non-hydrolyzable, orally active venous antithrombotic agents. AB - A (-)-conduritol F derivative was condensed with 4-ethyl-7-hydroxy-2H-1 benzopyran-2-one and converted into (+)-4-ethyl-7-[(1'R,2'S,3'S,4'R)-2',3',4'- trihydroxycyclohexyloxy]-2H-1-benzopyran-2-one ((+)-2). Enantiomer (-)-2 was obtained from a (+)-conduritol F derivative. The carbaxyloside (-)-2 with the L xylose configuration was more active than (+)-2 in the Wessler's model. PMID- 9873416 TI - Unusual synthesis of new glycine antagonists via sequential aldol condensation lactonization-elimination reaction. AB - Compounds 2 and 3 were designed in order to probe the North-East region of the strichnine-insensitive glycine binding site of the NMDA receptor. The two products were obtained readily by a tandem aldol condensation-lactonization elimination step which affords the desired E isomer with complete regioselection. PMID- 9873417 TI - Conodiparines A-D, new bisindoles from Tabernaemontana. Reversal of vincristine resistance with cultured cells. AB - Four new bisindoles of the vobasine-iboga type, conodiparines A-D were obtained from Tabernaemontana corymbosa which showed appreciable activity in reversing resistance in vincristine-resistant KB cells. PMID- 9873418 TI - Identification and SAR for a selective, nonpeptidyl thrombin inhibitor. AB - A novel, nonpeptidyl thrombin inhibitor, L-636,619 (1), was identified via topological similarity searching over the Merck Corporate Sample Database. X-ray crystallographic studies determined the geometry for ligand binding to the enzyme. Chemical modification of the P1 and P3 segments of the ligand resulted in enhanced potency and improvement in the chemical stability of the lead. Analog 9 proved to be the most interesting lead from this structurally novel series. PMID- 9873419 TI - Ascorbic acid-based inhibitors of alpha-amylases. AB - A series of ascorbic acid and isoascorbic acid derivatives has been evaluated as inhibitors of malt, bacterial, fungal, pancreatic and salivary alpha-amylases using a simple and quick assay procedure. The results demonstrate that the enediol moiety of ascorbic acid is essential for alpha-amylase inhibition. Acylation of the primary and secondary alcohols, and the absolute configuration of the secondary alcohol, do not affect the potency of inhibition. PMID- 9873420 TI - Synthesis of betulinic acid derivatives with activity against human melanoma. AB - Betulinic acid has been modified at C-3, C-20, and C-28 positions and the toxicity of the derivatives has been evaluated against cultured human melanoma (MEL-2) and human epidermoid carcinoma of the mouth (KB) cell lines. This preliminary investigation demonstrates that simple modifications of the parent structure of betulinic acid can produce potentially important derivatives, which may be developed as antitumor drugs. PMID- 9873421 TI - Peptide-based inhibitors of the hepatitis C virus serine protease. AB - Hexapeptide DDIVPC-OH is a competitive inhibitor of the hepatitis C virus (HCV) NS3 protease complexed with NS4A cofactor peptide. This hexapeptide corresponds to the N-terminal cleavage product of an HCV dodecapeptide substrate derived from the NS5A/5B cleavage site. Structure-activity studies on Ac-DDIVPC-OH revealed that side chains of the P4, P3 and P1 residues contribute the most to binding and that the introduction of a D-amino acid at the P5 position improves potency considerably. Furthermore, there is a strong preference for cysteine at the P1 position and conservative replacements, such as serine, are not well tolerated. PMID- 9873422 TI - C6 modification of the pyridinone core of thrombin inhibitor L-374,087 as a means of enhancing its oral absorption. AB - 1 (L-374,087) is a potent, selective, efficacious, and orally bioavailable thrombin inhibitor that contains a core 3-amino-2-pyridinone moiety. Replacement of the C6 pyridinone methyl group of 1 by a propyl group gave 5 (L-375,052), which retained all the excellent properties of 1, and also yielded higher plasma levels after oral dosing in dogs and rats. PMID- 9873423 TI - A free energy calculation can be used to predict K(+)-binding constants for new macrotetrolide antibiotics. AB - A free energy calculation technique was used to predict K+ binding constants for new macrotetrolides. The technique was validated by successfully predicting affinity constants for known, naturally produced, macrotetrolides. PMID- 9873424 TI - Korundamine A, a novel HIV-inhibitory and antimalarial "hybrid" naphthylisoquinoline alkaloid heterodimer from Ancistrocladus korupensis. AB - A unique heterodimeric naphthylisoquinoline alkaloid, korundamine A (2), comprised of two different monomeric biaryl halves, has been isolated from the Cameroonian tropical liana Ancistrocladus korupensis. Korundamine A is the first "hybrid" dimer found in the Ancistrocladaceae; in vitro, it demonstrated anticytopathic activity against HIV-1 and antimalarial activity against Plasmodium falciparum. PMID- 9873425 TI - Neuronorm is a potent and water soluble neurokinin A receptor antagonist. AB - We report here the synthesis and preliminary pharmacological characterization of a novel water soluble Neurokinin A receptor antagonist named Neuronorm. The synthesis was achieved in high yield by a combination of classical peptide synthesis methodologies, in both solution and solid phase. The pharmacological properties as neurokinin A receptor antagonist were assessed in in vitro experiments on rat vas deferens and guinea pig trachea, and were compared to those of MEN10627. PMID- 9873426 TI - Synthesis and hypoxia-selective cytotoxicity of a 2-nitroimidazole mustard. AB - A four-step synthesis of 5-[N,N-bis(2-chloroethyl)amino]-1-methyl-2 nitroimidazole from 1-methyl-2-nitroimidazole is described. This compound showed similar hypoxia-selective cytotoxicity to the dinitrobenzamide mustard SN 23,862 in UV4 cells (ca. 40-fold), and superior selectivity (> 7-fold) in repair competent AA8 cells. PMID- 9873427 TI - Heteroatom- and carbon-linked biphenyl analogs of Brequinar as immunosuppressive agents. AB - Structure-activity relationships were explored for some analogs of Brequinar having a linking atom between the 2-biphenyl substituent and the quinoline ring. Activities as inhibitors of dihydroorotate dehydrogenase and the mixed lymphocyte reaction were related to the overall shape and lipophilicity of the 2 substituent. PMID- 9873428 TI - The synthesis of symmetrical spermine conjugates using solid-phase chemistry. AB - The utility of spermine, selectively functionalised and immobilised on a solid support by means of the Wang "oxycarbonyl" linker is demonstrated by the solid phase synthesis of a number of spermine conjugates including the natural product and potent antihypertensive agent kukoamine. The synthesis opens up the area of solid-phase spermine chemistry and library generation based on the symmetrical spermine scaffold. PMID- 9873429 TI - Conformationally constrained analogues of diacylglycerol (DAG). 14. Dissection of the roles of the sn-1 and sn-2 carbonyls in DAG mimetics by isopharmacophore replacement. AB - The replacement of the sn-1 and sn-2 carbonyl esters in DAG-surrogate lactones by sulfonate esters showed that their isosteric properties in protein kinase C binding are controlled by the location of the hydrophobic alkyl chain on the molecule. The CO and SO2 groups appear to be true isosteres only when they are adjacent to the alkyl chain, which is presumed to insert normal to the lipid bilayer. PMID- 9873430 TI - Synthesis of a high-mannose-type glycopeptide analog containing a glucose asparagine linkage. AB - The title compound was prepared by enzymatic transfer of oligosaccharide to a synthetic pentapeptide containing the Glc-Asn linkage. The compound was not hydrolyzed by glycoamidases from plant and bacterial sources, but it inhibited both enzymes in the micromolar range. Its activity is compared to other potential inhibitors. PMID- 9873432 TI - Endothelin antagonists: discovery of EMD 122946, a highly potent and orally active ETA selective antagonist. AB - The discovery, in vitro and in vivo studies of the highly potent ETA antagonist EMD 122946 are presented. This compound displayed high binding affinity and functional antagonism [IC50 = 3.2 x 10(-11) M, pA2 = 9.5 (ETA)] and inhibited the ET-1 induced pressor response in pithed rats with an ED50 of 0.3 mg/kg. In conscious spontaneously hypertensive rats and in DOCA-salt hypertensive rats the compound lowered mean blood pressure with an ED50 of 0.06 mg/kg. EMD 122946 exhibited high bioavailability in rats and monkeys. PMID- 9873431 TI - The synthesis of new adenosine A3 selective ligands containing bioisosteric isoxazoles. AB - The synthesis and purinergic receptor binding of novel adenosine A3 ligands is described. Many selective A3 receptor agonists e.g. N-(3-iodobenzyl)adenosine-5' methyluronamide (IB-MECA) contain a 4'-ribosylalkylamide moiety. We found that this amide and other 4'-functional groups could be replaced with an isosteric isoxazole, and the target molecules retained potent binding to the recombinant human A3 receptor. PMID- 9873434 TI - Synthetic approaches to 2-(4-hydroxy-7-chromanyl)benzoic acids as antagonists of leukotriene B4. AB - Structural modification of 1 led to a series of 2-(4-hydroxy-7-chromanyl)benzoic acid LTB4 antagonists exemplified by 2 and 3. The use of an organostannane biaryl coupling, a non steroselective reduction and a chromatographic resolution limited the utility of this synthetic route. To address these issues, a new synthetic route was developed utilizing a palladium catalyzed coupling of aryl oxazolines in tandem with a stereospecific enone reduction as key synthetic steps. Resolution was achieved by fractional crystallization of a (S)-(-)-alpha methylbenzylamine salt. PMID- 9873433 TI - trans-3-Benzyl-4-hydroxy-7-chromanylbenzoic acid derivatives as antagonists of the leukotriene B4 (LTB4) receptor. AB - The SAR of a series of 2-(7-chromanyl)benzoic acids has been investigated with the aim of identifying potent and selective LTB4 receptor antagonists that maintain potency in complex biological fluids. We found optimal activity in derivatives with electron-withdrawing groups in the benzoic acid ring and with an unsubstituted C-3 benzyl group on the chromanol nucleus. While compounds containing a 3-(4-phenyl)benzyl chromanol substituent were potent LTB4 receptor antagonists, the increased lipophilicity imparted by the additional phenyl substituent led to decreased potency in the presence of plasma proteins. From among the potent compounds identified, CP-195543, the 5'-trifluoromethyl 3-benzyl chromanol, was selected for development. PMID- 9873436 TI - Synthesis and in vitro cytotoxicity of C(20)(RS)-camptothecin analogues modified at both B (or A) and E ring. AB - A series of C(7) and C(20)-substituted camptothecin derivatives (12-14, 16-18) are prepared. Their syntheses and in vitro cytotoxicities are reported. PMID- 9873435 TI - Development of new chromanol antagonists of leukotriene D4. AB - By addressing the issues of potency and metabolism in 3, a new series of LTD4 antagonists represented by (+)-26 was developed which is equipotent to clinical LTD4 antagonists Zafirlukast (1) and Pranlukast (2). PMID- 9873437 TI - Synthesis and antioxidant activity of novel amphipathic derivatives of tea polyphenol. AB - Hydrophobic derivatives of a tea polyphenol have been synthesized. 6, 8 Bis(octylthiomethyl)-epigallocatechin 3-O-gallate, 6, 8-bis(octylthiomethyl)-4 beta-(2-hydroxyethylthio)epigallocatechin 3-O-gallate and epigallocatechin 3-O-[4 O-(N-octadecylcarbamoyl)gallate] showed strong inhibition activity against lipid peroxidation of liposome caused by both lipid-soluble and water-soluble radical generators. PMID- 9873438 TI - Novel imidazole derivatives with subtype-selective antimuscarinic activity (1). AB - A series of N-substituted 4-amino-2,2-diphenylbutyramide derivatives was prepared as part of a search for subtype-selective antimuscarinic agents. The representative compound KRP-197, bearing a 2-methylimidazole ring as a surrogate of aliphatic amine, was found to be a highly potent and both M1- and M3-selective antimuscarinic agent. PMID- 9873439 TI - C-alkylated spiro[benzofuran-3(2H),4'-1'-methyl-piperidine-7-ols] as potent opioids: a conformation-activity study. AB - Among a series of C-alkylated analogs of the weak mu opioid ligand spiro[benzofuran-3(2H),4'-1'-methylpiperidine-7-ol] (1), the 2-methyl, 2-ethyl, and cis 3'-methyl analogs, namely compounds (+/-)2, (+/-)-3, and (+/-)-4, showed much enhanced mu-affinities, with (+/-)-4 being almost as potent as (-)-morphine; while the trans 3'-methyl analog (+/-)-5 remained a weak mu-binder. Energy calculations and nmr data indicated that compounds 2-4 favor phenyl-axial conformations, while compounds 1 and 5 favor phenyl-equatorial conformations. PMID- 9873440 TI - Modeling, synthesis and biological activity of novel antifungal agents (1). AB - Homology modeling of candida lanosterol C-14 demethylase, synthesis and in vitro antifungal activities of cyclohexyl analogs of restricticin are described. PMID- 9873441 TI - Synthesis of novel antifungal agents (2). AB - Synthesis of novel cyclohexyl analogs of restricticin using intramolecular radical cyclization and their in vitro and in vivo antifungal activities are described. PMID- 9873442 TI - Inhibition of lipid peroxidation mediated by indolizines. AB - Esters, ethers, carbonates and carbamates of 1-indolizinols and azaindolizinols exhibit a profound inhibition of lipid peroxidation in vitro. The antioxidants were prepared by cyclization of pyridines and diazines with diphenylcyclopropenone followed by introduction of the O-substituent. PMID- 9873443 TI - A potent and selective inhibition of parainfluenza 1 (Sendai) virus by new 6 oxiranyl-, 6-methyloxiranyluracils, and 4(3H)-pyrimidinone derivatives. AB - Several new 6-oxiranyl-, 6-methyloxiranyluracils, and pyrimidinone derivatives, synthesized by the lithiation-alkylation sequence of 1,3,6-trimethyluracil, 1,3 dimethyl-6-chloromethyluracil, and 2-alkoxy-6-methyl-4(3H)-pyrimidinones, showed a potent and selective antiviral activity against the parainfluenza 1(Sendai) virus replication. PMID- 9873444 TI - Reactive DNA. 2. Thioguanine used as a peg site for direct and specific introduction of biologically useful functional groups. AB - An efficient method is described for one-step and specific introduction onto thioguanine in DNA of a variety of biologically useful functional groups, including carboxymethyl, hydroxyethyl, aminoethyl groups and fluorescent tag. Reaction mechanisms were examined. The method could provide new tools for biochemical studies, such as DNA repair and sister chromatid exchange, and the method may be also applicable to RNA. PMID- 9873445 TI - Serine derived NK1 antagonists. 1: The effect of modifications to the serine substituents. AB - A series of novel serine derived NK1 antagonists is described. The effect of variations in the N-benzyl, O-benzyl and serine groups are used to define the elements which are necessary for binding. PMID- 9873446 TI - Serine derived NK1 antagonists. 2: A pharmacophore model for arylsulfonamide binding. AB - Modifications to the spirocyclic aryl sulfonamide portion of serine derived NK1 antagonists allow a partial pharmacophore model to be developed. PMID- 9873447 TI - A membrane-permeant, bioactivatable derivative of Ins(1,3,4)P3 and its effect on Cl(-)-secretion from T84 cells. AB - The synthesis of rac-2,5,6-tri-O-butyryl-myo-inositol 1,3,4-trisphosphate hexakis(acetoxymethyl) ester [Bt3-Ins(1,3,4)P3/AM, 1], a membrane-permeant derivative of myo-inositol 1,3,4-trisphosphate [Ins(1,3,4)P3] is reported. 1 inhibited calcium-mediated chloride secretion of T84 cells, suggesting a regulatory link of Ins(1,3,4)P3 and the biosynthesis of the known inhibitor myo inositol 3,4,5,6-tetrakisphosphate. PMID- 9873448 TI - Synthesis and anti-HIV activities of symmetrical N1,N3-dibenzyl-2-hydroxy-propane derivatives. AB - We report the synthesis and the anti-HIV activities of new C2-symmetrical and achiral N1,N3-dibenzyl-2-hydroxy-propane isosteres. Some of them showed significant inhibitory activity with respect to HIV-infected MT4 cells (compound 6a and 7a, IC50 = 0.1 microM). These new structurally simple compounds represent new synthons which can be suitable for combinatorial chemistry purposes. PMID- 9873449 TI - The synthesis and biological evaluation of a novel series of indole PDE4 inhibitors I. AB - This communication describes the synthesis and in vitro evaluation of a novel potent series of phosphodiesterase type (IV) (PDE4) inhibitors. The compounds described contain an indole moiety which replaces the 'rolipram-like' 3-methoxy-4 cyclopentoxy motif. Several of the compounds presented possess low nanomolar IC50's for PDEIV inhibition. In vivo activities determined from measurement of serum TNF-alpha levels in LPS challenged mice (mouse endotoxemia model) are also reported. PMID- 9873450 TI - Prodrug esters of the indolocarbazole CEP-751 (KT-6587). AB - Prodrug esters of the indolocarbazole CEP-751 (KT-6587) were prepared with the goal of identifying water soluble, stable but cleavable forms for intravenous dosing. A dipeptide proform Lys-beta-Ala (16, CEP-2563/KT-8391) was identified for advancement to clinical trials. PMID- 9873451 TI - Solid-phase synthesis of N-substituted amidinophenoxy pyridines as factor XA inhibitors. AB - An arylamidine linker has been employed for the solid-phase synthesis of N substituted amidinoaryloxypyridine analogs 2 via nucleophilic substitution on a fluoropyridyl template. Two novel N-substituted amidinoaryloxypyridine derivatives 2a and 2b were discovered via this approach. PMID- 9873452 TI - Novel cytokine release inhibitors. Part I: Triterpenes. AB - Tripterine and closely related triterpenoid derivatives as IL-1 beta release inhibitors are discussed. PMID- 9873453 TI - Inhibition of HIV integrase by novel nucleotides bearing tricyclic bases. AB - 5'-Monophosphates of several novel dideoxynucleosides bearing tricyclic nucleobases were synthesized. Both linear and angular ring-extended analogs of isomeric dideoxyadenosine 5'-monophosphate were discovered to have moderate to good inhibition of the viral-encoded enzyme, HIV integrase. The results suggest that the nucleotide binding site of HIV integrase can accommodate major modifications in the nucleobase, which is in stark contrast to the nucleotide binding site on HIV reverse transcriptase. PMID- 9873454 TI - 17 alpha-alkyl- or 17 alpha-substituted benzyl-17 beta-estradiols: a new family of estrone-sulfatase inhibitors. AB - A series of 17 alpha-derivatives of 17 beta-estradiol was synthesized and tested for their ability to inhibit the estrone-sulfatase activity transforming estrone sulfate to estrone. A strong inhibitory activity was obtained when an alkyl side chain or a substituted benzyl was introduced at position 17 alpha of estradiol. The 17 alpha-(3'-bromobenzyl)-estradiol (26) and 17 alpha-(4'-t-butylbenzyl) estradiol (30) were the most potent estrone-sulfatase inhibitors obtained in our study with IC50 values of 24 and 28 nM, respectively. They also represent a new family of estrone-sulfatase inhibitors. These compounds are about 300-fold more effective in interacting with the enzyme than the substrate estrone sulfate itself. PMID- 9873455 TI - Discovery of a novel benzyloxyisoquinoline derivative with potent anti Helicobacter pylori activity. AB - The synthesis and in vitro optimization of the anti-Helicobacter pylori activity of a novel series of benzyloxyisoquinoline derivatives discovered by a random screening process, are described. FR180102 (7f), having a 3-acetamido-2,6 dichlorobenzyl moiety, was found to have extremely potent activity against H. pylori and no effect against a series of common Gram-positive and Gram-negative bacteria. PMID- 9873456 TI - Efficient synthesis of 3'-glycosylated LacNAc-based oligosaccharides. AB - LacNAc-based oligosaccharides, including sialyl-(2-->3)-LacNAc, dimeric sialyl-(2 ->3)-LacNAc, trimeric sialyl-(2-->3)-LacNAc, beta-glucuronyl-(1-->3)-LacNAc, and 3-sulfo-beta-glucuronyl-(1-->3)-LacNAc, were synthesized efficiently from a single protected LacNAc derivative having both OH-3' and 4' unprotected. PMID- 9873457 TI - Syntheses of 2-[(3,5-dimethyl-4-methoxypyridyl)alkyl]-benzothiazolidine derivatives as a potential gastric H+/K(+)-ATPase inhibitor. AB - A series of 2-[(3,5-dimethyl-4-methoxypyridyl)alkyl]benzothiazolidine derivatives were synthesized and tested their inhibitory effects on gastric H+/K(+)-ATPase. Compound 4d exhibited potent in vitro inhibitory activity. PMID- 9873459 TI - [3H]LY341495, a highly potent, selective and novel radioligand for labeling Group II metabotropic glutamate receptors. AB - We report herein the synthesis and pharmacological characterization of a tritiated version of the potent and selective cyclopropyl amino acid LY341495 as a radioligand to label group II metabotropic glutamate receptors in rat brain homogenates. PMID- 9873458 TI - Nucleosides and nucleotides. 176. 2'-Deoxy-2'-hydroxylaminocytidine: a new antitumor nucleoside that inhibits DNA synthesis although it has a ribonucleoside structure. AB - The design and synthesis of potential antitumor antimetabolites 2'-deoxy-2' hydroxylaminouridine (2'-DHAU) and -cytidine (2'-DHAC) are described. We found that 2'-DHAC in neutral solution generated 2'-aminoxy radicals at room temperature. 2'-DHAC inhibited the growth of L1210 and KB cells, with IC50 values of 1.58 and 1.99 microM, respectively, more potently than 2'-DHAU, with IC50 values of 34.5 and 27.3 microM, respectively. 2'-DHAC was effective against 9 human cell lines, with IC50 values of in the micromolar range. The in vivo antitumor activity of 2'-DHAC was also examined using the mouse leukemia P388 model, which gave a T/C value 167%. Phosphorylation of 2'-DHAC by uridine/cytidine kinase was essential for its cytotoxicity, as suggested by a competition experiment using several common nucleosides. Inhibition of DNA synthesis was the predominant mechanism of action of 2'-DHAC, although it has a ribo-configuration. PMID- 9873460 TI - Novel inhibitors of bacterial two-component systems with gram positive antibacterial activity: pharmacophore identification based on the screening hit closantel. AB - This SAR study has shown that the salicylanilide is the pharmacophore for inhibition of the bacterial two-component system. Hydrophobic substituents improve the potency of inhibitors in this series; however, hydrophobicity is not the sole determinant for inhibition; structural and electronic requirements also exist. Closantel (1) was found to inhibit a two-component system and to have antibacterial activity against drug resistant S. aureus and E. faecium. PMID- 9873461 TI - SAR studies of diaryltriazoles against bacterial two-component regulatory systems and their antibacterial activities. AB - A series of diaryltriazole analogs was discovered to inhibit bacterial two component regulatory systems in our primary assays, KinA/Spo0F and NRII/NRI. They also showed inhibitory activity in whole cell mechanism-based assays, and they possessed potent activities against several strains of Gram-positive pathogenic bacteria in the standard MIC broth assay. PMID- 9873462 TI - 4-Alkylpiperidines related to SR-48968: potent antagonists of the neurokinin-2 (NK2) receptor. AB - A series of 4-alkylpiperidine derivatives related to the potent neurokinin-2 (NK2) receptor antagonist SR-48968 (1) is described. Simple aliphatic derivatives were found to be poorly active, but appropriate placement of an alcohol functional group afforded compounds that were of similar activity to 1. Several representatives in this series, such as the 4-(1-hydroxy-1-ethylpropyl)piperidine (14), were found to exhibit oral activity in a model of labored abdominal breathing in guinea pigs. These results expand the latitude of substituents available in this region of this series of NK2 receptor antagonists. PMID- 9873463 TI - Development of dual-acting agents for thromboxane receptor antagonism and thromboxane synthase inhibition. 2. Design, synthesis, and evaluation of a novel series of phenyl oxazole derivatives. AB - Synthesis and initial in vitro evaluation of a novel series of phenyl oxazole derivatives are described. An SAR study of the novel dual-acting TRA/TSI agent has revealed that the lipophilicity of the oxazole amide substituents greatly influences the TRA activity but not the TSI. The chain length of the alkenoic acid side chain affects both TRA and TSI. The optimal chain length for the combined activities was found to be n = 4 (heptenoic acid). PMID- 9873464 TI - A practical synthesis of 3-substituted delta 3,5(6)-steroids as new potential 5 alpha-reductase inhibitor. AB - A new and practical synthetic approach to 3-substituted delta 3,5(6)-Steroids, as potential 5 alpha-reductase inhibitor, is described. The key step involves Pd catalyzed coupling reaction of steroid 3-enol 5H-3-oxo octafluoropentanosulfonates. 3-Phenylacetylenyl substituted delta 3,5(6)-steroid 3g and 3-phosphate substituted delta 3,5(6)-steroid 3f in our synthesized compounds exhibited high 5 alpha-reductase inhibitory activity in vitro assay. PMID- 9873465 TI - Synthesis and structure-activity relationships of 2-pyridones: II. 8-(Fluoro substituted pyrrolidinyl)-2-pyridones as antibacterial agents. AB - The 8-position side chain of 2-pyridones is believed to be involved in the binding with bacterial DNA gyrase to form the ternary complex, making them very important for the activity of 2-pyridones. A series of 2-pyridones having fluoro substituted amines at the 8-position has been synthesized and their antibacterial activities and parmacokinetic properties are reported. PMID- 9873466 TI - Halohydrin analogues of cryptophycin 1: synthesis and biological activity. AB - The chloro-, bromo-, and iodo-derivatives 2-4 of the antimitotic drug cryptophycin 1 were synthesized by opening the epoxide ring. The biological activities of the compounds were tested in an in vitro microtubule assembly and a cell proliferation assay. The chloro-derivative 2 showed lower activity in the tubulin assay compared to 3 and 4, but they all showed similar inhibition in the proliferation assay. PMID- 9873467 TI - Peptidyl chemosensors incorporating a FRET mechanism for detection of Ni(II). AB - Hexapeptides incorporating two fluorophores flanking a tripeptide sequence that binds Ni(II) and Cu(II) with high affinity have been synthesized. While Cu(II) quenches the fluorescence of the resulting peptides, coordination of Ni(II) enables enhanced FRET (fluorescent resonance energy transfer) from one fluorophore to the other. PMID- 9873468 TI - Novel angucycline compound with both antigastrin- and gastric mucosal protective activities. AB - An angucycline series compound P371 A1 (1) from Streptomyces sp. P371 was established to have a novel structure comprising an ureido group at one of four sugar units on the basis of 2D NMR techniques. 1 exhibited an inhibitory activity against the pentagastrin-stimulated acid secretion as well as protective activities against HCl/ethanol- and indomethacin-induced gastric lesions. PMID- 9873469 TI - A novel series of 6-methoxy-1H-benzotriazole-5-carboxamide derivatives with dual antiemetic and gastroprokinetic activities. AB - A novel series of 6-methoxy-1H-benzotriazole-5-carboxamide derivatives with a medium perhydroazacycle ring in the amine moiety were prepared, and their antiemetic and gastroprokinetic activities were evaluated. Among them, N-(1 ethylhexahydroazepin-3-yl)-, N-(1-ethyloctahydroazocin-3-yl)- and N-(1 ethyloctahydroazonin-3-yl)-6-methoxy-1H-benzotriazole-5-carbo xamides (24, 36, 37) showed a potent antiemetic activity (inhibition of apomorphine-induced emesis in dogs) along with gastroprokinetic activity (gastric emptying in rats). PMID- 9873470 TI - Preparation of difluoro analogs of CCGs and their pharmacological evaluations. AB - All the stereoisomers of 2-(2-carboxy-3,3-difluorocyclopropyl)glycines (F2CCGs) were synthesized in enantiomerically pure forms using (R)-2,3-O-isopropyl ideneglyceraldehyde as a chiral precursor. L-F2CCG-I, one of the stereoisomers corresponding to an extended form of L-glutamate was found to be a potent agonist for metabotropic glutamate receptors (mGluRs). PMID- 9873471 TI - Synthesis and metabotropic glutamate receptor antagonist activity of N1 substituted analogs of 2R,4R-4-aminopyrrolidine-2,4-dicarboxylic acid. AB - A series of N1-substituted derivatives of (2R,4R)-4-aminopyrrolidine-2,4 dicarboxylate (2R,4R-APDC) has been prepared as constrained analogs of gamma substituted glutamic acids and examined for their effects at recombinant metabotropic glutamate receptor (mGluR) subtypes in vitro. Appropriate substitution of the N1 position of 2R,4R-APDC resulted in the identification of a number of selective group II mGluR antagonists. PMID- 9873472 TI - Identification and characterization of m4 selective muscarinic antagonists. AB - Our interest in the area of m4 muscarinic antagonists had led us to study a series of benzoxazine isoquinolines. One of the most potent and selective compounds of this series is example 1 with an IC50 value of 90.7 nM at m4 receptors, and 72-fold (m1), 38-fold (m2), 10-fold (m3), and 82-fold (m5) more selective compared to the other receptors. The synthesis and receptor binding affinity of analogs of 1 are reported. PMID- 9873473 TI - Asymmetric synthesis of antimitotic combretadioxolane with potent antitumor activity against multi-drug resistant cells. AB - The (S,S)-enantiomer of combretadioxolane (3), designed as a chirally preorganized derivative of combretastatin A-4, exhibited quite strong tubulin polymerization-inhibitory activity (IC50: 4-6 microM). (S,S)-3 is 20 times more potent than vincristine as an in vitro growth inhibitor (in terms of GI50) of the multi-drug-resistant (MDR) cell line PC-12, which produces P-glycoprotein. PMID- 9873474 TI - Actinomycin D, C2 and VII, inhibitors of Grb2-SHC interaction produced by Streptomyces. AB - Actinomycin D, C2 and VII, cyclic peptides, inhibited Grb2 SH2 domain association (IC50 5-7 microM) with a phosphotyrosine containing peptide derived from the Shc protein (pTyr317). Actinomycins are the first examples of nonphosphorylated natural ligands of SH2 domain. PMID- 9873475 TI - The novel cyclopropapyrroloindole(CPI) bisalkylators bearing 3,3'-(1,4 phenylene)diacryloyl group as a linker. AB - The novel cyclopropapyrroloindole(CPI) bisalkylators were synthesized and their antitumor activity was evaluated. Among these derivatives, AT-760 (5a) in which the two 3-methoxycarbonyl-2-trifluoromethylCPI (MCTFCPI) moieties are connected with a 3,3'-(1,4-phenylene)diacryloyl group, was found to exhibit more prominent cytotoxicity and antitumor activity than U-77,779 (bizelesin) (1). PMID- 9873476 TI - Diastereoselective two-directional synthesis and cation transport ability of the central tristetrahydrofuranyl unit of meso polyether glabrescol as naturally occurring podand. AB - The diastereoselective synthesis of the central 2,5-linked tristetrahydrofuran (trisTHF) 6 of naturally occurring meso polyether, glabrescol (5), has been achieved in a two-directional manner by the vanadium(V)-catalyzed anti oxidative cyclizations of diol 14. The trisTHF podand 6 and its stereoisomeric analogs 7 and 8 exhibited outstanding cation transport abilities for physiologically important Na+ and K+. PMID- 9873477 TI - General construction pattern of histamine H3-receptor antagonists: change of a paradigm. AB - Novel omega-phenyl substituted and unsubstituted alkyl and alkenyl imidazole derivatives were prepared and tested for their antagonist activity in vitro and in vivo at histamine H3-receptors. Some compounds showed high in vitro and in vivo H3-receptor activity despite their structure bearing no polar moiety in the centre of the molecule which is a common structural feature of all other antagonists known. Quite probably there are further in vivo effects for some compounds resulting from other receptor interactions. PMID- 9873478 TI - Reaction of guanosine with glucose under oxidative conditions. AB - Incubation of glucose with guanosine under oxidative conditions at 37 degrees C or 70 degrees C leads to the formation of a main product, which can be detected by HPLC/DAD. The compound was isolated and identified as N2-carboxymethyl guanosine (CMG). To confirm the structure, CMG was also synthesized from glyoxal and guanosine. PMID- 9873479 TI - Design of novel porphyrin-binding peptides based on antibody CDR. AB - Novel porphyrin-binding peptides were designed on the basis of an antigen binding site of an antiheme monoclonal antibody. Synthetic peptides were modified with a pyrene moiety. The spectroscopic measurements revealed that the synthetic peptides bound a porphyrin effectively. PMID- 9873480 TI - Syntheses of potent Leu-enkephalin analogs possessing beta-hydroxy-alpha,alpha disubstituted-alpha-amino acid and their characterization to opioid receptors. AB - Novel Leu-enkephalin (Leu-Enk) (1) analogs possessing various types of alpha substituted serine instead of its glycine residue in the position 2 were synthesized via an efficient O,N-migration method. The binding characteristics of the synthetic analogs using Chinese hamster ovary (CHO) cells expressed cloned rat mu-, delta-, and kappa-receptors revealed that [(1R,2S)-Ahh2]Enk (7) was the most potent agonist of delta-opioid receptors among all the synthetic analogs tested, and was 10 times more potent than the native Leu-Enk. PMID- 9873481 TI - SAR of novel biarylmethylamine dopamine D4 receptor ligands. AB - SAR for a novel series of dopamine D4 receptor ligands is shown. Very selective, highly potent compounds like 1-(2-pyrimidinyl)-4-(3-(3-thienyl)-benzyl) piperazine (5f) and 2-(4-(1-fluorenylmethyl)-1-piperazinyl)-pyrimidine (8c) were obtained. PMID- 9873482 TI - Increased HLA-DQ2-affinity of a synthetic gliadin peptide by acid-induced deamidation of glutamine residues. AB - Presentation of antigenic gliadin peptides by the HLA-DQ2 molecule is considered as a key event in celiac disease pathogenesis. Chemical deamidation of the side chains of glutamine residues might have a strong influence on gliadin peptide binding to the DQ2 molecule. Glutamine deamidation of A-gliadin peptide (45-56) under acidic conditions corresponding to the gastric environment was studied using RP-HPLC, Edman degradation, capillary electrophoresis and electrospray mass spectrometry. Deamidation resulted in peptides with increased DQ2-affinities as assessed in a cell-free binding assay. PMID- 9873483 TI - Synthesis and antibacterial in vitro activity of novel analogues of nematophin. AB - The synthesis and in vitro antibacterial activity of new derivatives and analogues of nematophin are described. It was shown that the unsubstituted amide NH-group is essential for bioactivity. Alkyl- or arylsubstitution at the 1 position results in a distinct increase of antibacterial activity. Addition of protein (blood or serum) to the culture media reduces the inhibitory activity on bacteria. PMID- 9873484 TI - A synthetic acceptor substrate for Trypanosoma brucei UDP-Gal: GPI anchor side chain alpha-galactosyltransferases. AB - A synthetic analogue of a trisaccharide fragment of the Trypanosoma brucei Variant Surface Glycoprotein (VSG) GlycosylPhosphatidyInositiol (GPI) anchor, Gal alpha-1,3(Man-alpha-1,6)-Man-alpha-O-octyl (1), serves as a substrate for two T. brucei alpha-galactosyltransferases. The principle tetrasaccharide product derived from (1) contains a Gal-alpha-1,2-Gal linkage. PMID- 9873485 TI - Design and synthesis of thiol-reactive lipopeptides. AB - Lipopeptides are potent adjuvants that trigger an immune response against covalently conjugated low molecular mass antigens. We report here the design and synthesis of thiol-reactive lipopeptides (6, 7) which can be incorporated into liposomes and react, under mild conditions, with synthetic peptides carrying a thiol function. PMID- 9873486 TI - Thromboxane modulating agents. 4. Design and synthesis of 3-(2-[[(4 chlorophenyl)sulfonyl]-amino]ethyl)benzenepropanoic acid derivatives as potent thromboxane receptor antagonists. AB - The design of a series of thromboxane receptor antagonists based on 3-(2-[[(4 chlorophenyl)sulfonyl]amino]ethyl)benzenepropanoic acid (1) is described. Addition of an arylmethyl group at the 5-position of 1 gave exceptionally potent agents in vitro and in vivo, with 13a (UK-147,535) giving complete blockade of the TxA2 receptor for greater than 12 hours in dogs, following an oral dose of 0.1 mg/kg. PMID- 9873487 TI - Pyrazolo[1,5-a]pyrimidine CRF-1 receptor antagonists. AB - A series of 3-phenylpyrazolo[1,5-a]pyrimidines was prepared and found to have affinity for the human CRF-1 receptor. The 3-dimensional structure of one of the most potent analogs in this series, 10d, was determined by X-ray crystallography and suggests the spatial requirements for potent CRF-1 receptor binding affinity in this series. PMID- 9873488 TI - Kampanols: novel Ras farnesyl-protein transferase inhibitors from Stachybotrys kampalensis. AB - Farnesyl-protein transferase (FPTase) is a critical enzyme that participates in the post-translational modification of the Ras protein. Inhibitors of this enzyme have the potential of being novel anticancer agents for tumors in which the ras oncogene is found mutated and contributes to cell transformation. Continued screening of natural product extracts led to the isolation of kampanols, which are novel and specific inhibitors of FPTase. The most active kampanols exhibited IC50 values between 7 to 13 microM against human recombinant FPTase. The isolation, structure determination, and biological activity of these compounds are described. PMID- 9873489 TI - Design and synthesis of conformationally-constrained MMP inhibitors. AB - A novel series of conformationally constrained matrix metalloprotease inhibitors was identified. The potencies observed for these inhibitors were highly dependent upon the substitution pattern on the caprolactam ring as well as the succinate moiety. PMID- 9873490 TI - Biological properties of synthetic glycoconjugate mimics of heparin comprising different molecular spacers. AB - The in vitro antithrombotic activity of synthetic glycoconjugates I and II, comprising a flexible polyethylene glycol type and a rigid polyglucose type spacer, respectively, are compared to heparin. PMID- 9873491 TI - The design, synthesis, and structure-activity relationships of a series of macrocyclic MMP inhibitors. AB - A series of succinate-derived hydroxamic acids incorporating a macrocyclic ring were designed, synthesized, and evaluated as inhibitors of matrix metalloproteinases. The inhibitors were designed based on the published X-ray crystal structure of batimastat (1) complexed with human neutrophil collagenase (MMP-8). The synthesized compounds were shown to inhibit selected MMPs in vitro with low nanomolar potency. PMID- 9873492 TI - Antineoplastic agents. 397: Isolation and structure of sesterstatins 4 and 5 from Hyrtios erecta (the Republic of Maldives). AB - The wide ranging marine sponge Hyrtios erecta is the source of the spongistatins, a new class of macrocyclic lactone antineoplastic agents. Continuation of a detailed investigation of cancer cell growth inhibitory (P388 lymphocytic leukemia) fractions (trace) from H. erecta has revealed the presence (10(-5) to 10(-7)% yield) of cytotoxic pentacyclic sesterterpenes. Employing P388 leukemia and human tumor cell line-guided bioassay techniques, two new moderate inhibitors of cancer cells were isolated and named sesterstatins 4 (1a, P388 ED50 4.9 micrograms/mL) and 5 (1b, DU-145 prostate GI50 1.9 micrograms/mL). Similar to other sesterterpenes, sesterstatin 5 inhibited growth of a Gram-positive bacterium. High field (500 MHz) 2-D NMR techniques were primarily employed for initial structural assignments, and structural assignments were confirmed by X ray crystal structure determination of sesterstatin 4 (1a) and 5 (1b). PMID- 9873493 TI - Synthesis and pharmacological evaluation of benzofuran-acetamides as "antineophobic" mitochondrial DBI receptor complex ligands. AB - A sries of novel benzofuran analogues of N,N-di-n-hexyl-2-phenylindole-3 acetamide (5, FGIN-1-27), a potent and highly specific mitochondrial DBI receptor complex ligand, were synthesized by a modified Fischer method and found in vitro and in vivo to be equally potent and selective as FGIN-1-27. PMID- 9873494 TI - Effects of phosphorothioate oligodeoxyribonucleotide and oligoribonucleotides on human complement and coagulation. AB - We have synthesized and studied the effects of phosphorothioate (PS) oligodeoxyribonucleotide (DNA) and oligoribonucleotides (RNA, 2'-O-methyl-RNA and 2'-5'-RNA) on complement activation and prolongation of activated partial thromboplastin time (aPTT) in vitro. These results suggest that a PS-DNA prolongs aPTT, and inhibits complement lysis more than do the PS-RNA analogs. PMID- 9873495 TI - Identification of N2-(1-carboxymethyl)guanine (CMG) as a guanine advanced glycation end product. AB - The reaction of D-glucose with 9-methylguanine produces N2-fructosyl-9 methylguanine, which undergoes oxidation to produce N2-(1-carboxymethyl)-9 methylguanine as a major adduct. PMID- 9873496 TI - 3-Pyridyloxypropanolamine agonists of the beta 3 adrenergic receptor with improved pharmacokinetic properties. AB - Pyridyloxypropanolamines L-749,372 (8, beta 3 EC50 = 3.6 nM) and L-750,355 (29, beta 3 EC50 = 13 nM) are selective partial agonists of the human receptor, with 33% and 49% activation, respectively. Both stimulate lipolysis in rhesus monkeys (ED50 = 2 and 0.8 mg/kg, respectively), with minimal effects on heart rate. Oral bioavailability in dogs, 41% for L-749,372 and 47% for L-750,355, is improved relative to phenol analogs. PMID- 9873497 TI - Synthesis of novel substituted pyridines as inhibitors of endothelin converting enzyme-1 (ECE-1). AB - A series of bi-aryl pyridine carboxylic acids has been prepared and evaluated as inhibitors of ECE-1. The analogs were prepared by Pd catalyzed cross couplings of halogenated pyridines with heteroaryl organo-boranes, -tinate or -zincate derivatives. PMID- 9873498 TI - Probing the importance of spacial and conformational domains in captopril analogs for angiotensin converting enzyme activity. AB - A new synthesis of 4,5-methano-L-prolines and the enzymatic activity of the corresponding N-(3-mercapto-2-R-methyl-propionyl) analogs as inhibitors of angiotensin converting enzyme are described. PMID- 9873499 TI - The structure-activity relationships of a series of suicide inhibitors of phospholipase A2. AB - A series of mechanism-based inhibitors of phospholipase A2 (SIBLINKS) were synthesized. These new SIBLINKS are phospholipid analogues that contain a para substituted phenyl 3,3-dimethylglutaryl group in the place of the sn-2 acyl chain. The effect of the phenyl leaving group on inhibitory activity was studied by varying the electron-withdrawing ability of the para-substituted group. A strong correlation was observed between the leaving group potential of the suicide inhibitor and the inhibitory activity of the derivative toward cobra venom phospholipase A2. PMID- 9873501 TI - Solid phase synthesis of oligomannopeptoids that mimic the concanavalin A-binding trimannoside. AB - Oligomannopeptoids from the dimer to the hexamer were produced by solid phase synthesis and their abilities to bind to concanavalin A (ConA) were assessed. The assessment indicated similarity between the oligomannopeptoids and the naturally occurring oligomannosides in the enthalpy of the binding and the valence number vs binding strength relationship, encouraging the use of the oligomannopeptoids as oligomannoside mimics. PMID- 9873500 TI - The cholesterol metabolite cholest-4-en-3-one and its 3-oxo derivatives suppress body weight gain, body fat accumulation and serum lipid concentration in mice. AB - Based on the findings that cholest-4-en-3-one, an intestinal metabolite of cholesterol, has an anti-obesity effect on animals, the structure-effect relationship of its 3-oxo derivatives and related compounds were investigated. Cholesten-3-ones, which possesses an enone structure with a carbonyl group at C3, markedly inhibit body weight gain and body fat accumulation, as well as the levels of serum triglyceride and cholesterol in animals without any clinical abnormalities. PMID- 9873502 TI - Synthesis and biological activity of 2-methyl-20-epi analogues of 1 alpha,25 dihydroxyvitamin D3. AB - Synthesis and biological evaluation of all eight possible A-ring diastereomers of 2-methyl-20-epi-1,25-dihydroxyvitamin D3 are described. Among the analogues synthesized. 2 alpha-methyl-20-epi-1 alpha,25-dihydroxyvitamin D3 exhibited exceptionally high potency. The double modification of 2-methyl substitution and 20-epimerization yielded analogues with unique activity profiles. PMID- 9873503 TI - Potent inhibition of protein-tyrosine phosphatase-1B using the phosphotyrosyl mimetic fluoro-O-malonyl tyrosine (FOMT). AB - To enhance PTP binding interactions, both inside and outside the pTyr binding pocket, a thioether-cyclized peptide has been designed based on the EGF receptor autophosphorylation sequence (EGFR988-993) "Asp-Ala-Asp-Glu-pTyr-Leu", in which the pTyr resiude has been replaced by the nonphosphorus-containing pTyr mimetic fluoro-O-malonyltyrosine (FOMT, 2). The resulting peptide 4 exhibits a Ki value of 170 nM, making it one of the most potent inhibitors of PTP1B yet reported. PMID- 9873504 TI - Anti-AIDS agents. 33. Synthesis and anti-HIV activity of mono-methyl substituted 3',4'-di-O-(-)-camphanoyl-(+)-cis-khellactone (DCK) analogues. AB - Four isomeric methyl substituted DCK analogues (2-5) were asymmetrically synthesized from different starting materials. 3-Methyl, 4-methyl, and 5-methyl 3',4'-di-O-(-)-camphanoyl-(+)-cis-khellactone (2-4) all were extremely potent against HIV-1 replication in H9 lymphocyte cells with EC50 and therapeutic index values of < 4.23 x 10(-7) microM and > 3.72 x 10(8), respectively, which are much better than those of DCK and AZT in this assay. PMID- 9873505 TI - Novel H3 receptor antagonists. Sulfonamide homologs of histamine. AB - Sulfonamides derived from 4(5)-(omega-aminoalkyl)-1H-imidazoles containing chain lengths of three- to five-carbons were synthesized. Good to moderate H3 receptor binding affinities were observed for several butyl and pentyl homologs, whereas binding affinities were considerably weaker in the propyl series. Separation of the imidazole ring and the sulfonamide unit by a four- or five-carbon tether afforded potent H3 receptor antagonists. PMID- 9873506 TI - Novel imidazole derivatives with subtype-selective antimuscarinic activity (2). AB - A series of 4-(2-methylimidazol-1-yl)-2,2-diphenylbutyramide derivatives was prepared as part of a search for subtype-selective antimuscarinic agents. On the basis of measurements of the antimuscarinic activity and subtype-selectivity for M2 and M3 muscarinic receptors, the structure-activity relationships of these compounds are discussed. PMID- 9873507 TI - A dipyrido [2,3-b:3',2'-f]azepine analog of the HIV-1 reverse transcriptase inhibitor nevirapine. AB - The syntheses of 11-ethyl and 11-cyclopropyldipyrido[2,3-b:3',2'-f]azepines, analogs of the HIV-1 reverse transcriptase inhibitor nevirapine 1, are described. These compounds exhibit potency equivalent to nevirapine in the inhibition of wild-type and some mutant RT enzymes. PMID- 9873508 TI - Conformationally restricted analogues of nicotine and anabasine. AB - A series of conformationally restricted analogues of nicotine has been synthesized and evaluated as agonists of neuronal acetylcholine receptors. Compound 2 (SIB-1663), which selectively activated human recombinant alpha 2 beta 4 and alpha 4 beta 4 nAChRs, was shown to be active in animal models of Parkinson's disease and pain. PMID- 9873509 TI - In vitro anti-human immunodeficiency virus (HIV) activity of the chromanone derivative, 12-oxocalanolide A, a novel NNRTI. AB - The three chromanone derivatives, (+)-, (-)-, and (+/-)-12-oxocalanolide A (2), were evaluated for in vitro antiviral activities against HIV and simian immunodeficiency virus (SIV). The compounds were determined to be inhibitors of HIV-1 reverse transcriptase (RT) and exhibited activity against a variety of viruses selected for resistance to other HIV-1 nonnucleoside RT inhibitors. They are the first reported calanolide analogues capable of inhibiting SIV. PMID- 9873510 TI - Synthesis and structure-activity relationships of 7-(2 aminoalkyl)morpholinoquinolones as anti-Helicobacter pylori agents. AB - A series of the titled compounds was synthesized and tested for anti-Helicobacter pylori activities. We discovered Y-34867 having the most potent activity against Helicobacter pylori among the quinolones tested along with high photostability. Furthermore, Y-34867 showed an excellent therapeutic effect in the experimental Helicobacter pylori infected Mongolian gerbil model. PMID- 9873511 TI - Hepatoprotective and nitric oxide production inhibitory activities of coumarin and polyacetylene constituents from the roots of Angelica furcijuga. AB - The methanolic extract from the roots of Angelica furcijuga KITAGAWA was found to exhibit protective effects on liver injury induced by D-galactosamine (D-GalN) and lipopolysaccharide (LPS). From the methanolic extract, seventeen coumarins, two phenylpropanoids, and two polyacetylenes were isolated and examined their in vitro and in vivo hepatoprotective effects and inhibitory activity of NO production in macrophages. A acylated khellactone, isoepoxypteryxin, showed protective activity against D-GalN-induced cytotoxicity in primary cultured rat hepatocytes. On the other hand, six acylated khellactones (hyuganins A, B, C, and D, anomalin, isopteryxin) and two polyacetylenes [(-)-falcarinol and falcarindiol] strongly inhibited NO production induced by LPS in cultured mouse peritoneal macrophages, and also other acylated khellactones (isoepoxypteryxin, pteryxin, and suksdorfin) and a coumarin glycosides (praeroside II) were found to show the activity. By comparison of the inhibitory activities for acylated khellactones with those for other coumarins, acyl groups were found to be essential to exerting potent activity. PMID- 9873512 TI - Reactions of 7- and 9-aminoadenines with 2,4-pentanedione. Formation of new ring systems, pyridazino[6,1-f]purine and pyridazino[1,6-e]purine. AB - Reactions of 7-aminoadenines with 2,4-pentanedione yielded pyridazino[6,1 f]purines and 5-pyrazolylpyrimidines. The structure of the latter was identified by X-ray analysis. Reaction of 9-amino-N6-methyladenine with 2,4-pentanedione gave pyridazino[1,6-e]purine and Schiff base products. Possible reaction mechanisms are discussed. PMID- 9873513 TI - Isoxazolylthioamides as potential immunosuppressants a combinatorial chemistry approach. AB - A library of thioamide derivatives of leflunomide 1a and of its bioactive metabolite 1b has been synthesised on solid phase. Thus, para-substituted phenylacetic acids were coupled to TentaGel and were subsequently reacted with aromatic isothiocyanates. Treatment of the resulting enaminothioamides with hydroxylamine led to their simultaneous cyclisation and cleavage from the resin affording 2-25. Their in vitro profiling demonstrated that the amide-thioamide isologous substitution was detrimental of the biological activity. PMID- 9873514 TI - Requirement of the glycosyl parts in platycodin D to stimulate pancreatic exocrine secretion. AB - The whole structure of platycodin D is found to be essential to stimulate the volumetric increase in the pancreatic exocrine secretion, while the prosapogenins prepared from platycodin D increased only protein output of pancreatic juice. PMID- 9873515 TI - Structure-based design of N-[2-(1-piperidinylethyl)]-N'-[2-(5-bromopyridyl)] thiourea and N-[2-(1-piperazinylethyl)]-N'-[2-(5-bromopyridyl)]-thiourea as potent non-nucleoside inhibitors of HIV-1 reverse transcriptase. AB - A novel computer model of the HIV reverse transcriptase (RT) non-nucleoside inhibitor (NNI) binding pocket, which was generated using high resolution crystal structure information from 9 individual RT/NNI complexes, revealed previously unrecognized ligand derivatization sites for phenethylthiazolylthiourea (PETT) derivatives. Spatial gaps surrounding the pyridyl ring of the active PETT derivative trovirdine were discovered during modeling procedures. Docking studies using the computer-generated model of the binding pocket (composite binding pocket) suggested that the replacement of the planar pyridyl ring of trovirdine with a nonplanar piperidinyl or piperazinyl ring, which occupy larger volumes, would better fill the spacious Wing 2 region of the butterfly-shaped NNI binding pocket. The anti-HIV activity of the synthesized heterocyclic compounds N-[2-(1 piperidinylethyl)]-N'-[2-(5-bromopyridyl)]-thiourea and N-[2-(1 piperazinylethyl)]-N'-[2-(5-bromopyridyl)]-thiourea was examined in HTLVIIIB infected peripheral blood mononuclear cells. Both compounds were more potent than trovirdine and abrogated HIV replication at nanomolar concentrations without any evidence of cytotoxicity. PMID- 9873516 TI - The first analogues of LNA (locked nucleic acids): phosphorothioate-LNA and 2' thio-LNA. AB - LNA (Locked Nucleic Acids, 1, X = O, Y = O) is a novel oligonucleotide analogue capable of recognizing complementary DNA and RNA with unprecedented thermal affinities. Synthesis of the first chemically modified LNA analogues is reported. A 9-mer phosphorothioate-LNA containing three LNA thymine monomers (1, X = O, Y = S, Base = thymin-1-yl) and 9-mer LNAs containing one, three or five 2'-thio-LNA monomers (1, X = S, Y = O, Base = uracil-1-yl) were able to recognize both complementary DNA and RNA with thermal affinities comparable to those of parent LNA. PMID- 9873517 TI - Enantioselective synthesis and pharmacology of 11-hydroxy-(1'S,2'R) dimethylheptyl-delta 8-THC. AB - An enantioselective synthesis of the (1'S,2'R)-dimethylheptyl cannabinoid side chain has been developed and employed in the synthesis of 11-hydroxy-(1'S,2'R) dimethylheptyl-delta 8-THC (3). Pharmacology, in vivo and in vitro, indicate (3) to be one of the most potent traditional cannabinoids known. PMID- 9873518 TI - Synthesis of a paclitaxel isomer: C-2-acetoxy-C-4-benzoate paclitaxel. AB - A synthesis of the C-2-acetoxy-C-4-benzoate paclitaxel 2 is described. This analog has the substituents at C-2 and C-4 transposed. The key steps in the synthesis include the sequential use of Red-Al as reducing agent for the regioselective reduction of the C-2 benzoate and the C-4 acetoxy within the baccatin core. Iso-paclitaxel 2 was considerably less potent than paclitaxel in tubulin polymerization and in vitro cytotoxicity assays. PMID- 9873519 TI - A simple procedure for solid-phase synthesis of peptide nucleic acids with N terminal cysteine. AB - Problems were encountered during attempts to prepare N-terminal cysteine substituted peptide nucleic acids (PNAs) from commercially available, Fmoc protected monomers. These problems have been surmounted by the use of an S-t butylmercapto protecting group on the cysteine moiety. The solid-phase syntheses are carried out via a simplified procedure which should be generally useful for manual PNA synthesis. PMID- 9873520 TI - Design, synthesis, and biological activity of novel purine and bicyclic pyrimidine factor Xa inhibitors. AB - The synthesis of amidinoaryloxy 9-benzyl-8-methyl-9H-purine, 7,8-dihydropteridine 6(5H)-one and 5,7-dihydropyrimido[4,5-b][1,4]oxazine-6-one inhibitors of Factor Xa is described. These compounds show nanomolar potency against FXa and maintain high selectivity over thrombin and trypsin. PMID- 9873521 TI - Discovery of IRL 3461: a novel and potent endothelin antagonist with balanced ETA/ETB affinity. AB - IRL 3461, N-butanesulfonyl-[N-(3,5-dimethylbenzoyl)-N-methyl-3-[4-(5-+ ++isoxazolyl) -phenyl]-alanyl]-(L)-valineamide, a potent and bifunctional (ETA + ETB) [Ki(ETA) = 1.8 nM, Ki(ETB) = 1.2 nM] antagonist was discovered by structural modification of IRL 2500, an ETB selective antagonist. IRL 3461 was found to be stable on incubation with human, rat, mouse, and guinea pig plasmas. PMID- 9873522 TI - Stereoselective synthesis of a novel and bifunctional endothelin antagonist, IRL 3630. AB - IRL 3630 (3), a single enantiomer of IRL 3461 with more potency was identified. Coupling reaction of the racemic fragment (1) with the chiral (L) valinesulfonamide (2) under a biphasic solvent system (CH2Cl2-H2O) successfully led to the predominant formation of the desired isomer (3) with concomitant isomerization of 1. IRL 3630, N-butanesulfonyl-[N-(3,5-dimethylbenzoyl)-N-methyl 3-[4-(5-+ ++isoxazolyl) -phenyl]-(D)-alanyl]-(L)-valineamide, is a highly potent and bifunctional (ETA + ETB) antagonist [Ki(ETA) = 1.5 nM, Ki(ETB) = 1.2 nM]. PMID- 9873524 TI - L-tryptophan urea amides as NK1/NK2 dual antagonists. AB - We report that a systematic modification of an NK1 receptor selective antagonist resulted in the identification of novel compounds, 4c and 4d, with high affinity for both NK1 and NK2 receptors. PMID- 9873523 TI - C32-O-imidazol-2-yl-methyl ether derivatives of the immunosuppressant ascomycin with improved therapeutic potential. AB - A series of C32-O-aralkyl ether derivatives of the FK-506 related macrolide ascomycin have been prepared based on an earlier reported C32-O-cinnamyl ether design. In the present study, the nature of the aryl tethering group was varied in an attempt to improve oral activity. An imidazol-2-yl-methyl tether was found to be superior among those investigated and has resulted in an ascomycin analog, L-733,725, with in vivo immunosuppressive activity comparable to FK-506 but with an improved therapeutic index. PMID- 9873525 TI - 4-[(1H-imidazol-4-yl) methyl] benzamidines and benzylamidines: novel antagonists of the histamine H3 receptor. AB - A series of amidine substituted phenyl-, benzyl-, and phenethylimidazoles based on the known H3 agonist SK&F 91606 (4) has been synthesized and tested as ligands for the histamine H3 receptor. Insertion of a phenyl ring between the imidazole ring and the amidine moiety produces antagonists. The benzyl series was found to be the most potent and was further investigated. Compounds 9c and 18 (entries 5 and 12, Table 1) are potent ligands for the H3 receptor with K(i) values of 16 nM and 7.2 nM respectively. In vivo, both compounds were shown to be equipotent to thioperamide (2), the standard H3 antagonist. PMID- 9873526 TI - Alkyl side-chain derivatives of sordaricin as potent antifungal agents against yeast. AB - Sordarin (1) was converted to 5 and 6, which showed potent antifungal activity against yeast. A series of C1-C9 alkyl side-chain derivatives was prepared, from which it was found that the optimal activity occurred with C5. A comparison of side chains with different unsaturation showed that the cis-alkene was the most active. This result suggested that the folding of the side chains might be crucial for the optimal activity. PMID- 9873527 TI - Combinatorial chemistry: from peptides and peptidomimetics to small organic and heterocyclic compounds. AB - Modified dipeptides have been used successfully for the generation of a variety of small organic and heterocyclic combinatorial libraries, including linear urea, polyamine, hydantoin, thiohydantoin, cyclic urea, cyclic thiourea and bicyclic guanidine. The synthesis and screening results for a number of these libraries are described. The solid phase synthesis of heterocyclic compounds such as diazepine and thiomorpholinone are also described. PMID- 9873528 TI - Comparison of the intrinsic kinase activity and substrate specificity of c-Abl and Bcr-Abl. AB - We studied the intrinsic tyrosine kinase activity and substrate specificity of c Abl and Bcr-Abl protein tyrosine kinases (PTKs) using the peptide substrates discovered from a synthetic combinatorial peptide library. Our data indicate that the phosphorylation of these peptides by Bcr-Abl was consistently stronger than that by c-Abl. Bcr-Abl also showed substrate preference towards those peptides with one or more positive charges. PMID- 9873529 TI - In vitro selection of phosphorothiolated aptamers. AB - A pool of RNA molecules that contained exclusively phosphorothioate internucleoside linkages was used as a starting point for the selection of aptamers that bind to basic fibroblast growth factor (bFGF), and appear to act as heparin mimics. PMID- 9873530 TI - A combinatorial approach to chemical modification of subtilisin Bacillus lentus. AB - The reaction between methanethiosulfonate reagents and cysteine mutants of subtilisin is quantitative and can be used to prepare chemically modified mutant enzymes (CMMs) with novel properties. The virtually unrestricted structural variations possible for CMMs presents a preparative and screening challenge. To address this, a rapid combinatorial method for preparing and screening the activities of CMMs has been developed. PMID- 9873532 TI - Exploring structure-activity relationships around the phosphomannose isomerase inhibitor AF14049 via combinatorial synthesis. AB - Phosphomannose Isomerase (PMI) has been shown by genetic methods to be an essential enzyme in fungal cell wall biosynthesis. The PMI inhibitor AF14049 was discovered as an unanticipated side product from high-throughput library screening against the enzyme from C, albicans. Solid-phase synthetic methods were developed and a series of libraries and discrete analogs synthesized to explore SAR around AF14049. PMID- 9873531 TI - Novel inhibitors of alpha 4 beta 1 integrin receptor interactions through library synthesis and screening. AB - A library of 2302 small molecule beta-turn mimetics was screened for inhibition of the alpha 4 beta 1 integrin-CS1 splice variant binding interaction. Preliminary data revealed several active ligands, and validation with purified material culminated in the identification of some of the first small molecule ligands (1, IC50 = 5 microM, and 2, IC50 = 8 microM) to be reported for this class of integrins. PMID- 9873533 TI - Rapid optimization of an ICE inhibitor synthesis using multiple reaction conditions in a parallel array. AB - Optimization of a 2-step reaction sequence was accomplished in 3-4 days, with over 200 different reaction conditions evaluated. Combinatorial arrays were performed using the optimized conditions to synthesize 590 new compounds which were tested for inhibition against N-His (D381E) ICE. Thirty-five compounds showed at least a tenfold improvement in activity compared to an initial standard. PMID- 9873534 TI - Identification of potent inhibitors of Plasmodium falciparum plasmepsin II from an encoded statine combinatorial library. AB - An encoded 13,020-member combinatorial library was synthesized containing a statine core. Evaluation of this library with plasmepsin II, an aspartyl protease required for hemoglobin metabolism in the malaria parasite, led to the identification of potent and selective inhibitors as well as novel structure activity relationships. PMID- 9873535 TI - Highly efficient and versatile synthesis of libraries of constrained beta-strand mimetics. AB - The general approach of using a bicyclic template to produce inhibitors of the protease superfamily of enzymes has been investigated. The Diels Alder cycloaddition reaction on solid support has been found to be highly efficient for the synthesis of libraries of compounds that mimic the beta-strand secondary structure of proteins. Several potent and selective inhibitors of proteases have been discovered. PMID- 9873536 TI - Tight-binding streptavidin ligands from a cyclic peptide library. AB - During the screening of a soluble library of cyclo(AXXXXXAE)K-CONH2, a cyclic peptide cyclo(AHPQFPAE)K-CONH2 was identified as a tight-binding ligand (IC50 = 128 nM) and found to bind 1000-fold more tightly than its linear peptide to streptavidin. The results of this study suggest that library screening of conformationally constrained cyclic peptides can be an effective means for the discovery of high affinity ligands. PMID- 9873537 TI - Synthesis of sialyl Lewis X mimetics using the Ugi four-component reaction. AB - Application of the Ugi four-component condensation to rapidly synthesize a library of glycopeptide mimics of the tetrasaccharide SLe(x) as inhibitors of E- and P-selectin, and to study the effect of varied functionality in mimics on the inhibition is described. PMID- 9873538 TI - Higher order iminodiacetic acid libraries for probing protein-protein interactions. AB - Higher order iminodiacetic acid diamide trimer (560 compounds) and tetramer libraries (1260 compounds) are described and were assembled in a convergent multistep solution-phase synthesis for use in studying protein-protein interactions. PMID- 9873539 TI - New piperazinyl polyazacyclophane scaffolds, libraries and biological activities. AB - Four novel unsymmetric piperazinyl polyazacyclophane scaffolds 1-4 were synthesized in high yields by an efficient cyclization strategy. Twenty-six libraries 12-37 (total 16,000 compounds) were generated by a solution-phase combinatorial approach from 1-4 and thirty-eight functional groups. Potent antibacterial and HIV-1 tat/TAR protein-RNA disrupting activities were discovered. PMID- 9873540 TI - "Cut and combine": an easy membrane-supported combinatorial synthesis technique. AB - A combinatorial synthesis process involving sequential cycles of cutting a membrane support into pieces and combining these into groups and subjecting the groups to simultaneous solid-phase chemical reactions is demonstrated by the rapid assembly of four hundred N-terminally biotinylated, soluble, octameric peptide pools. Index patterns printed onto the synthesis membrane allowed a direct identification of the compounds. These were used to study protein kinase substrate selection in a parallel microplate adapted 32P-phosphorylation assay with subsequent spotting on a biotin-capture membrane. PMID- 9873541 TI - Deconvolution by omission libraries. AB - Omission libraries, synthesized by omitting one amino acid in all coupling positions, are very efficient tools for the rapid identification of the amino acid components of bioactive peptides. Based on the determined amino acids, an occurrence library can be defined and prepared which is much less complex than the full one while still comprising the bioactive peptide. PMID- 9873542 TI - Investigating highly crosslinked macroporous resins for solid-phase synthesis. AB - The washing efficiencies of a chromophore and the reaction rates of a classical esterification reaction are improved with macroporous resins (MRs) relative to a classical Merrifield resin. Furthermore, Wacker-oxidation of a MR bound alkene yielded the expected methylketone product whereas an alkene bound to a low crosslinked Merrifield resin gave no product, a function of the relative permeability of each of these resins to the aqueous solvent conditions employed. PMID- 9873543 TI - Solid-phase synthesis of a library of piperazinediones and diazepinediones via Kaiser oxime resin. AB - A combinatorial library of piperazinediones has been prepared by automated parallel solid-phase synthesis. The five-step reaction protocol makes use of Kaiser oxime resin, to enable cleavage from the polymeric support concomitant with an intramolecular displacement reaction, under very mild conditions. The methodology was also successfully extended to the preparation of the seven membered ring homologs, diazepinediones. PMID- 9873544 TI - Combinatorial chemistry of hydantoins. AB - Access to combinatorial chemistry of hydantoins is provided by convenient and versatile methods for the solid phase synthesis of libraries of 3,5-, 1,3- and 1,3,5-substituted hydantoins. The preparation of trisubstituted hydantoins features a Mitsunobu reaction for introduction of the substituent on N-1. PMID- 9873545 TI - Hantzsch pyrrole synthesis on solid support. AB - An efficient method for solid-phase synthesis of pyrroles is described. Polystyrene Rink amide resin is acetoacetylated and converted into polymer bound enaminones upon treatment with primary amines. These then undergo a Hantzsch reaction with alpha-bromoketones to yield pyrroles. After cleavage with 20% trifluoroacetic acid in dichloromethane pyrrole-3-carboxamides are obtained in excellent purity. PMID- 9873546 TI - Chemically-tagged Mitsunobu reagents for use in solution-phase chemical library synthesis. AB - A general method for high-throughput product purification of Mitsunobu reactions is described. Tagged phosphine and azodicarboxylate reagents are used to synthesize individual library members in solution-phase. Workup and purification are easily accomplished by post-reaction sequestration of the tagged reagents and reagent byproducts by a complementary functionalized ion exchange resin. The reagents are utilized in a 3 step library synthesis. PMID- 9873547 TI - A mixed resin bed for the quenching and purification of tetrabutylammonium fluoride mediated desilylating reactions. AB - A polymer-bound calcium sulfonate resin is prepared and used to sequester tetrabutylammonium fluoride. The simultaneous use of the calcium sulfonate resin with a sulfonic acid resin is used for the quenching and purification of desilylating reactions involving tetrabutylammonium fluoride as the reagent. Employment of this resin workup technique eliminates the need for a liquid-phase extractive protocol allowing the procedure to be easily automated. PMID- 9873548 TI - Construction of a family of biphenyl combinatorial libraries: structure-activity studies utilizing libraries of mixtures. AB - A set of biphenyl aminoacid building blocks has been synthesized. These were used to construct partially-peptidic combinatorial libraries as equimolar multi component samples. Activity of members of this library as vitronectin receptor antagonists is described, together with SAR studies of the most active members. These studies illustrate several important features of combinatorial libraries. PMID- 9873549 TI - A simple method for coupling aldehydes to solid support. AB - A new method for attaching aldehydes to solid supports has been developed employing a 2,2-bis(hydroxymethyl)propionic acid (DMPA) functionalized resin. High loading levels are obtained for both aryl and alkyl aldehydes protected as their respective acetals. Treatment of the derivatized resin with 95% TFA then cleanly affords the recovered aldehyde in high yield. PMID- 9873550 TI - "Propylene spaced" allyl tin reagents: a new class of fluorous tin reagents for allylations under radical and metal-catalyzed conditions. AB - A new generation of propylene-spaced fluorous allyltin reagents [(Rf(CH2)3)3SnCH2CH = CH2] is described. These succeed in radical allylations where their lower homologs (ethylene-spaced) fail, and they provide improved performance in transition metal catalyzed allylations. The reagents and byproducts are readily separated by simple fluorous-organic liquid-liquid or solid-liquid extractions. PMID- 9873551 TI - High throughput purification of combinatorial libraries. AB - This article summarizes recent advances at MDS Panlabs which provide for the high throughput preparative HPLC purification of our Optiverse screening library. Topics covered include unique methods for the preparation, purification, characterization, and plating of purified screening compounds. Also discussed are procedures for data tracking as samples proceed through the purification process. PMID- 9873552 TI - Synthesis and structure-activity relationship of substituted 1,2,3,4 tetrahydroisoquinolines as N-type calcium channel blockers. AB - Voltage Activated Calcium Channel (VACC) blockers have been demonstrated to have utility in the treatment of pain and stroke. A series of aminomethyl substituted isoquinolinol derivatives with potent functional activity for N-type VACC's have been identified. Their synthesis and preliminary pharmacology are discussed herein. PMID- 9873553 TI - A photoactivated prodrug. AB - A photolabile derivative (1) of the anticancer drug, 5-fluorodeoxyuridine (2), was designed and synthesized as a model prodrug. Photolysis of 1 with long wavelength UV light rapidly released 2 in solution. While compound 1 alone is nontoxic to cells, the presence of both 1 and UV irradiation (lambda = 350 nm) resulted in potent inhibition of cell growth. PMID- 9873554 TI - NMR and topochemical studies of peptidomimetic HIV-I protease inhibitors containing a cis-epoxide amide isostere. AB - NMR and topochemical studies of irreversible HIV-1 protease inhibitors containing a cis-epoxide as amide isostere have been carried out to identify conformational preference of the inhibitors in solution. The inhibitors prefer to adopt extended conformations similar to the beta-strand in solution. PMID- 9873555 TI - Synthesis of 6-O-methyl-azithromycin and its ketolide analogue via Beckmann rearrangement of 9(E)-6-O-methyl-erythromycin oxime. AB - The synthesis of 6-O-methyl-azithromycin and its aza-ketolide analogue have been achieved by carrying out the Beckmann rearrangement of the readily available 9(E) 6-O-methyl-erythromycin oxime 1. In contrast to the C14 ketolides like HMR 3647, the aza-ketolide turns out to be inactive, thus demonstrating that the addition of a 3 keto function and ring expansion, from 14 to 15 membered ring, could be deleterious for the antibacterial activity. PMID- 9873556 TI - Design and synthesis of analogs of vitamin E: antiproliferative activity against human breast adenocarcinoma cells. AB - Analogs of alpha-tocopherol (vitamin E; compounds 3-9) have been synthesized and tested for their antiproliferative activity using the human breast cancer cell line, MCF7. Compounds 6-9 were synthesized from a common symthom, rac-Trolox (14) and are soluble/miscible at physiological pH. Compounds 4, 8, and 9 were found to have antiproliferative activity at micromolar concentrations. PMID- 9873557 TI - Soluble polymer traceless linker investigations: solvent effects on the desulfonylation of polyethylene glycol (PEG) substituted aryl alkyl sulfones with sodium amalgam. AB - The soluble polymer polyethylene glycol containing a traceless aryl alkyl sulfone linker was found to be resistant to reductive cleavage with sodium amalgam (Na/Hg). Detailed investigations revealed that the solvent plays an important role in the desulfonylation reaction; subsequently it was determined that MeOH/DMF (1/8) is an ideal solvent system for the cleavage of this traceless linker. PMID- 9873558 TI - Solid-phase synthesis of potential protein tyrosine phosphatase inhibitors via the Ugi four-component condensation. AB - A library of 108 alpha,alpha-difluoromethylenephosphonic acids was prepared by Ugi four-component condensation using Rink-NH2 resin, 4-[(diethoxyphosphinyl) difluoromethyl]benzoic acid, and a set of 18 aldehydes and 6 isonitriles. Following resin cleavage, the diethylphosphonate esters were hydrolyzed with trimethylsilyl bromide to yield the free acids which were assayed for inhibition of PTP alpha, PTP beta and PTP epsilon. PMID- 9873559 TI - Synthesis and biological evaluation of two analogues of (S)-alpha-methyl-3 carboxyphenylalanine. AB - Two analogues 2, 3 of (S)-alpha-methyl-3-carboxylphenylalanine 1 were synthesized to test their activity for metabotropic glutamate receptors (mGluRs). Both compounds 2 and 3 were inactive as antagonist for mGluRs, but 2 showed weak agonistic activity for GluR6 in contrast to that reported by Kemp and co-workers. PMID- 9873560 TI - N-carbamoyl analogs of Zafirlukast: potent receptor antagonists of leukotriene D4. AB - Exploration of the indole nitrogen region of Zafirlukast (1) has uncovered a potent series of cysteinyl leukotriene D4 (LTD4) antagonists. These studies showed that a variety of functionality could be incorporated in this region of the molecule without sacrificing potency. Efforts to exploit this site in order to improve oral efficacy are discussed. PMID- 9873561 TI - Functional characteristics of a series of N4-substituted 1-(2,3-dihydro-1,4 benzodioxin-5-yl)piperazines as 5-HT1A receptor ligands. Structure-activity relationships. AB - The agonistic/antagonistic profile of a series of 10 N4-substituted 1-(2,3 dihydro-1,4-benzodioxin-5-yl)piperazines is evaluated in the in vitro adenylyl cyclase assay. The profile is severely affected by the characteristics of the N4 substituents ranging from full agonism (benzamidoethyl derivative 1), mixed agonism/antagonism (phthalimidobutyl derivative 7) to predominantly antagonism (saccharinbutyl derivate 9). A novel full antagonist 10, as potent as WAY 100635, is obtained by substitution of Cl at C-7 of the benzodioxinyl moiety in 9. PMID- 9873562 TI - Templates for design of inhibitors for serine proteases: application of the program DOCK to the discovery of novel inhibitors for thrombin. AB - The program DOCK was used to search for novel inhibitors for alpha-thrombin. Four among the top twelve best scoring compounds from the Cambridge Structural Data Base inhibited this enzyme, and three of them inhibited alpha-thrombin in a competitive mode. These molecules are expected to serve as general templates for structural elaboration in targeting diverse serine proteases for selective inhibition. PMID- 9873563 TI - Design and synthesis of N-alkylated saccharins as selective alpha-1a adrenergic receptor antagonists. AB - Benign prostatic hyperplasia can be managed pharmacologically with alpha-1 adrenergic receptor antagonists. Agents that demonstrate selectivity for the alpha-1a receptor subtype may offer advantages in clinical applications with respect to hypotensive side effects. The N-alkylated saccharins reported here represent a new class of subtype selective alpha-1a adrenergic receptor antagonists which demonstrate potent effects on prostate function in vivo and are devoid of blood pressure side effects. PMID- 9873564 TI - Synthesis and effects of novel thiazole derivatives against thrombocytopenia. AB - 5-(2-Pyridylsulfonyl)-2-thiazolamine (2) was effective both in mitomycin C (MMC) induced thrombocytopenia and in an animal model of idiopathic thrombocytopenic purpura (ITP). It also suppressed the increase of autoantibodies against platelets in the ITP model and showed no blood toxicity. Chemical modification of 2 led to the discovery of more potent compounds against MMC-induced thrombocytopenia. PMID- 9873565 TI - H-phosphonate derivatives as novel peptide deformylase inhibitors. AB - Peptide deformylase catalyzes the removal of the N-terminal formyl group from nascent polypeptides during prokaryotic protein maturation and is essential for bacterial survival. Its absence from eukaryotic organisms makes it an attractive target for designing novel antibacterial agents. Peptidyl H-phosphonates were synthesized and shown to be competitive inhibitors of the deformylase. PMID- 9873566 TI - QSAR study of the peptidic fibrinogen inhibitors FK633, FR158999 and related derivatives, using a novel and useful hydrophobic descriptor (logPmw). AB - A QSAR study using the novel hydrophobic descriptor (logPmw), which is a descriptor for membrane affinity, of our fibrinogen inhibitors FK633 (1), FR158999 (21), and related derivatives was performed, and resulted in good correlation (n = 19, s = 0.268, F = 6.38**, r = 0.667). Based on these results, we constructed a hypothesis by which these potent inhibitors bind to the receptor via the biomembrane and the C-terminal moiety functions as an anchor moiety. PMID- 9873567 TI - The development of novel and selective p56lck tyrosine kinase inhibitors. AB - Early T-cell receptor mediated signal transduction involves the activation of several tyrosine protein kinases. One of these tyrosine kinases, p56lck, is expressed primarily in T-cells and Natural Killer (NK) cells and has been shown to be critical for their proliferative and effector functions. Indandiones have been identified as a potent and selective chemical class that inhibits p56lck. PMID- 9873568 TI - Selective alpha-1a adrenergic receptor antagonists. Effects of pharmacophore regio- and stereochemistry on potency and selectivity. AB - The anti-anxiety agent ipsapirone has been shown to have modest affinity for alpha-1 receptors. We disclose the discovery of potent alpha-1a receptor subtype selective antagonists based on the ipsapirone structure which possess selectivity versus the 5-HT receptors tested. These antagonists were obtained by tethering a saccharin ring to 4-phenyl-3-carboxyethyl piperidines. The design principles which led to this structural motif are discussed. The synthesis of key analogs, their SAR, as well as results of selected in vitro and in vivo studies are described. PMID- 9873569 TI - Design and construction of novel thrombin inhibitors featuring P3-P4 quaternary lactam dipeptide surrogates. AB - Potent serine protease inhibitor 1a featuring a hybrid P3-P4 quaternary lactam dipeptide surrogate was prepared based upon SAR and molecular modeling investigations and in order to further probe the S2/S3 thrombin and FXa subsites. An efficient and concise synthetic route to the key aminolactam intermediate 4 was developed. The design, synthesis, and biological activity of this target and its P3-P4 diastereomer 1b is presented. PMID- 9873570 TI - Naphthoquinone analogs as inactivators of cdc25 phosphatase. AB - cdc25A and cdc25B were significantly overexpressed in certain types of cancers, and they represent potential therapeutic targets for anticancer drug. In this study, naphthoquinone analogs as cdc25A phosphatase inactivators were investigated. PMID- 9873571 TI - 5'-Peptidyl substituents allow a tuning of the affinity of oligodeoxyribonucleotides for RNA. AB - The affinity of amide-linked 5'-aminoacyl and 5'-dipeptidyl DNA octamers for two RNA undecamers with 3'-overhangs was measured via UV melting analysis. A sequence dependent increase in melting points was observed. At low ionic strength, two appended lysine residues elevate melting points more than two additional A:U base pairs. PMID- 9873572 TI - Design of novel tripeptides with macrophage migration-enhancing activity. AB - A new approach to the generation of biologically active peptides from non-peptide lead was demonstrated. C,N-termini protected Phe-Phe-His derivatives possessing macrophage migration-enhancing activity were designed on the basis of the structure of imidazoline derivative 1, and these structure-activity relationships were also described. PMID- 9873573 TI - Inhibitors of farnesyl protein transferase. Synthesis and biological activity of amide and cyanoguanidine derivatives containing a 5,11-dihydro[1]benzthiepin, benzoxepin, and benzazepin [4,3-b]pyridine ring system. AB - Bioisosteric replacement of the C-6 carbon atom in piperidine I and piperazine II with S, O, and N heteroatoms is described. Amide and cyanoguanidine derivatives of these compounds were evaluated in vitro and found to be good inhibitors of farnesyl-protein transferase. An improved method of preparing the 5,11-dihydro [1]-benzthiepin nucleus 6 was accomplished in high yield and with excellent regioselectivity using an AlCl3 melt protocol. PMID- 9873574 TI - Dibasic benzo[b]thiophene derivatives as a novel class of active site directed thrombin inhibitors. 2. Exploring interactions at the proximal (S2) binding site. AB - In an effort to increase the thrombin inhibitory activity of a novel series of inhibitors (i.e., 1a), substituents were incorporated at the C-3" position of the C-3 aryl ring (2). Consistent with the X-ray crystallography studies, small hydrophobic groups at the C-3" site (Br and Me) enhanced thrombin inhibitory activity by 8-fold. However, a few more hydrophilic substituents (NO2 and OMe) also enhanced the potency of the series. The biological results are discussed in terms of molecular modeling studies. PMID- 9873575 TI - Modulation of apoptosis in human lymphocytes by adenosine analogues. AB - We previously described that 2-chloroadenosine (2CA) and 2-chloro-2' deoxyadenosine (2CdA) induced apoptosis in human peripheral blood mononuclear cells (PBMC). In this study we tested different adenosine analogues on PBMC and we found that the modifications introduced in the 2CA structure prevented the molecule from exerting its apoptotic effect. On the other hand, substitutions on 2CdA are tolerated, although with a significant decrease in activity. PMID- 9873576 TI - Solid-phase stereoselective synthesis of 2'-O-methyl-oligoribonucleoside phosphorothioates using nucleoside bicyclic oxazaphospholidines. AB - The use of 2'-OMe-ribonucleoside bicyclic oxazaphospholidines derived from (R)- or (S)-2-pyrrolidinemethanol has enabled the stereoselective synthesis of (Rp)-, and (Sp)-2'-O-methyloligoribonucleoside phosphorothioates. Interestingly, higher stereoselectivity (96-98%) was observed in the synthesis of (Sp)-2'-O-methyl oligoribonucleoside phosphorothioates compared to that in the case of (Sp) oligodeoxyribonucleoside phosphorothioates (90%). PMID- 9873577 TI - A new spacer group derived from arylmalonaldehydes for glucuronylated prodrugs. AB - A new glucuronylated prodrug of doxorubicin, potentially useful for ADEPT or PMT cancer chemotherapy, has been prepared from 4-methyl phenyl malonaldehyde. The enol ether spacer, linked via a carbamate to the 3'-amino group of doxorubicin is rapidly cleaved after beta-glucuronidase (E coli) catalyzed hydrolysis at pH 7.2 and 37 degrees C. PMID- 9873578 TI - Substrate specificity of thermostable farnesyl diphosphate synthase with alkyl group homologs of isopentenyl diphosphate. AB - 3-Alkyl group homologs of isopentenyl diphosphate were examined for the reactivity as substrates of the thermostable farnesyl diphosphate (FPP) synthase of Bacillus stearothermophilus. Even 3-n-propyl- and 3-n-butyl-but-3-enyl diphosphates, which are hardly acceptable by animal FPP synthases, are accepted by this bacterial enzyme as substrates to react with dimethylallyl- and geranyl diphosphates, yielding 7-methyl-3-n-propylocta-2,6-dienyl- and 7,11-dimethyl-3-n propyldodeca-2,6,10-trienyl diphosphate, respectively. PMID- 9873579 TI - Two novel oleanolic acid saponins having a sialyl Lewis X mimetic structure from Achyranthes fauriei root. AB - Two novel triterpene glycosides, achyranthosides E and F, were isolated as methyl esters from the root of Achyranthes fauriei, an antiinflammatory medicinal plant. Their structures were characterized as oleanolic acid glucuronides having unique substituents composed of C6H9O5 and C9H15O7, respectively, at the C-3 position of glucuronic acid. These compounds are active components which can inhibit the excess recruiting of neutrophiles to injured tissues 1,000 times more potently than sialyl Lewis X. PMID- 9873580 TI - Evidence supporting the activity of 2'-C-cyano-2'-deoxy-1-beta-D-arabino pentafuranosylcytosine as a terminator in enzymatic DNA-chain elongation. AB - To investigate the stability of 2'-C-cyano-2'-deoxy-1-beta-D-arabino pentafuranosylcytosine 3'-phosphoric acid, its thymidine ester was prepared via the phosphoramidite method using allyl protection for the phosphate function. This ester is stable under acidic conditions but extremely labile under basic conditions, decomposing with a cleavage of the internucleotide bond even at pH 7.3. PMID- 9873581 TI - Benzylamine-based selective and orally bioavailable inhibitors of thrombin. AB - A series of p-aminomethylphenylalanine derivatives were investigated as novel thrombin inhibitors. This study led to potent inhibitors of thrombin (Ki up to 3.3 nM) that are trypsin-selective, highly orally bioavailable in rats, and highly permeable across Caco-2 cells. The P1 benzylamine binding mode in the thrombin active site was identified by X-ray crystallographic analysis. PMID- 9873582 TI - Synthesis of 13C labelled daidzein and formononetin. AB - Efficient methods are described for the synthesis of daidzein and formononetin labelled with a single 13C atom at the 4-position, to prepare material for metabolic studies. PMID- 9873583 TI - Synthesis of boronic acid analogs of L-arginine as alternate substrates or inhibitors of nitric oxide synthase. AB - The asymmetric synthesis of an unprotected alpha-amino boronic acid analog of L arginine 1a (boroarg-OH. 2 HCl) and its N alpha-acetyl derivative 1b (Ac-boroarg OH. HCl) is described. These compounds were evaluated as substrates and inhibitors of recombinant nitric oxide synthases (NOS). Boroarg-OH 1a selectively inhibited inducible NOS (IC50 = 50 microM) compared to the neuronal isoform (IC50 = 300 microM). PMID- 9873584 TI - New antifungals selected by molecular topology. AB - Molecular topology has been applied to find the new lead antimycotic compounds. Among the selected compounds stands out 3,3'-(4,4'-Biphenylene)bis(2,5-diphenyl 2H-tetrazolium chloride), Benztropine mesylate and Dicyclopentamethylenethiuram disulphide, with minimum inhibitory concentrations between 1.6 and 2 micrograms/mL. PMID- 9873585 TI - N alpha-alkylated derivatives of 2-phenylhistamines: synthesis and in vitro activity of potent histamine H1-receptor agonists. AB - New potent N alpha-alkylated histamine H1-receptor agonists have been prepared and functionally evaluated for partial agonist potency and selectivity. N alpha Methyl-2-(3-trifluoromethylphenyl)histamine contracts ileal segments and aortic rings of guinea-pig with a relative potency of 174% (95% confid. lim. 161-188%) and 217% (164-287%), respectively (histamine: 100%) and is the most potent H1 receptor agonist described so far. PMID- 9873586 TI - PD 176252--the first high affinity non-peptide gastrin-releasing peptide (BB2) receptor antagonist. AB - In this paper we describe the development of a novel series of non-peptide, "balanced" neuromedin-B preferring (BB1)/gastrin-releasing peptide preferring (BB2) receptor ligands as exemplified by PD 176252. PD 176252, which exhibits nanomolar affinity for both the BB1 (Ki = 0.15 nM) and BB2 (Ki = 1.0 nM) receptors, has been demonstrated to be a competitive antagonist at these bombesin receptor subtypes. PMID- 9873588 TI - General synthesis of 3-substituted alkenyl GABA as potential anticonvulsants. AB - Stereospecific synthesis of cis and trans 3-substituted vinyl-gamma-aminobutyric acid analogs were obtained by either a Claisen rearrangement or a Wittig reaction from common diene precursors. PMID- 9873587 TI - In situ generated O-glycan core 1 structure as substrate for Gal(beta 1-3)GalNAc beta-1,6-GlcNAc transferase. AB - beta-Galactosidase from bovine testes was used in a one pot reaction together with a recombinant beta-1,6-GlcNAc transferase for the synthesis of GlcNAc(beta 1 6)GalNAc(alpha 1-OBn) (core 6-Bn). The galactosidase, which reversibly links galactose via a (beta 1-3) linkage to N-acetylgalactosamine, provides the substrate for the GlcNAc transferase in situ. The synthesis was carried out with a yield > 90%. PMID- 9873589 TI - Synthesis and binding of stable bisubstrate ligands for phosphoglycerate kinase. AB - Stable bisubstrate ligands of phosphoglycerate kinase (PGK) have been synthesized with AMP or ADP conjugated to hydrolytically-stable, symmetrical analogues of 1,3 bisphosphoglycerate and their binding to yeast PGK evaluated. Their Kds decrease with net negative charge, with a penta-anionic analogue 7 showing highest affinity-in accordance with its approximation to the transition state for the reaction catalysed by PGK. PMID- 9873591 TI - Mechanism based representation of the active site of 5 alpha-reductase (5AR). AB - In the present study, we have attempted to determine a detailed representation of the 5 alpha-Reductase (5AR) active site involving the elucidation of the transition state for the steroid delta 4 reduction reaction (the 'NADPH substrate' complex), onto which steroidal and non-steroidal inhibitors were superimposed. We conclude that: (i) there is a requirement for groups to mimic the steroid substrate A-ring; (ii) the area about C(3), C(4), C(5) and C(6) of T appears to be sterically hindered, and; (iii) the area of the active site about the C(17) of the steroid substrate does not possess hydrogen bonding groups and is not restricted. PMID- 9873590 TI - Synthesis and properties of prodrugs activated in hypoxia to give bleomycin analogues. AB - Prodrugs bioreductively activated to bleomycin analogues are reported. The production of hydroxyl radicals in the presence of FE(II) and dioxygen by both the prodrugs and the activated products are determined and their in vitro cytotoxicity measured. PMID- 9873592 TI - Biaryl acids: novel non-nucleoside inhibitors of HIV reverse transcriptase types 1 and 2. AB - A series of biaryl acids has been found to show micromolar inhibition of the HIV reverse transcriptase (RT) from types 1 and 2 with IC50S in the micromolar range. The series was discovered by consideration of the polymerase active site and sub structure searching of the company compound collection. Synthesis of analogues to investigate the SAR is described. Two of these compounds have shown inhibition of HIV-2 RT only. PMID- 9873593 TI - Synthesis and evaluation of a novel series of phosphodiesterase IV inhibitors. A potential treatment for asthma. AB - The synthesis and pharmacological profile of a novel series of potent and selective phosphodiesterase type IV (PDE IV) inhibitors is described. PMID- 9873594 TI - Aryl sulfonamides as selective PDE4 inhibitors. AB - A series of novel selective phosphodiesterase 4 (PDE4) inhibitors has been developed which displays activity both in vitro and in vivo. These compounds possess good selectivity for the catalytic site of PDE4 over the high affinity Rolipram binding site. In vivo studies demonstrate a reduced propensity to display the emetic side effects which are commonly observed with PDE4 inhibitors. PMID- 9873595 TI - Potassium peroxymonosulfate (oxone)--an efficient oxidizing agent for phosphothio compounds. AB - Potassium peroxymonosulfate (oxone) is demonstrated as a versatile chemoselective and stereospecific oxidizing agent for phosphothio compounds. Its application in nucleotide chemistry is presented. PMID- 9873596 TI - P2-proline-derived inhibitors of calpain I. AB - The syntheses and biological activities of a series of calpain I inhibitors, derived from D- and L-Pro, are described. PMID- 9873597 TI - Novel antineoplastic agents with efficacy against multidrug resistant tumor cells. AB - A novel series of pentafluorobenzenesulfonamides has been shown to inhibit the growth of a variety of human tumor cell lines. Among the cell types against which these agents were evaluated were the multidrug resistant (MDR) cell lines MCF 7/ADR and P388/ADR. The cytotoxic activity of members of this series of compounds was not affected by the multidrug resistant pump in MCF-7/ADR or P388/ADR cells. PMID- 9873598 TI - The synthesis and biological activity of a novel series of diazepine MMP inhibitors. AB - A novel series of diazepine-based hydroxamic acid inhibitors of MMP-1, MMP-9, and MMP-13 were prepared and evaluated both in vitro and in vivo. PMID- 9873599 TI - The importance of GLU361 position in the reaction catalyzed by cholesterol oxidase. AB - Cholesterol oxidase stereospecifically isomerizes cholest-5-en-3-one to cholest-4 en-3-one. When the base catalyst for isomerization, Glu361, is mutated to Asp, the rate of deprotonation of cholest-5-en-3-one is not affected, but protonation of the dienolic intermediate becomes rate-limiting. This may be a consequence of the large distance between the catalytic base and carbon-6 of the intermediate in the mutant enzyme. PMID- 9873600 TI - Thalidomide analogs and PDE4 inhibition. AB - N-Phthaloyl 3-amino-3-arylpropionic acid analogs of thalidomide that are potent inhibitors of tumor necrosis factor-alpha are reported. These compounds were found to be potent inhibitors of phosphodiesterase 4. PMID- 9873601 TI - New generation dopaminergic agents. 5. Heterocyclic bioisosteres that exploit the 3-OH-N1-phenylpiperazine dopaminergic template. AB - The synthesis of several bioisosteric analogs based on the 3-OH-N1 phenylpiperazine dopamine D2 agonist template (i.e., 4) is described. The indolone (5) and 2-CF3-benzimidazole (13) were observed to have excellent affinity for the D2 receptor. Several D4 selective compounds were also identified. Molecular modeling studies and a putative bioactive conformation are discussed. PMID- 9873602 TI - A high affinity, mu-opioid receptor-selective enkephalin analogue lacking an N terminal tyrosine. AB - We report a high affinity, mu opioid receptor selective enkephalin analogue in which the N-terminal tyrosine residue thought to be required for such high affinity is replaced by phenylalanine. The high affinity can be traced to a shift of the ligand's N-terminal residue within the mu receptor binding pocket, which diminishes the importance of the usual hydrogen bond between the tyrosine phenolic moiety and the receptor. PMID- 9873603 TI - Design of a high affinity peptidomimetic opioid agonist from peptide pharmacophore models. AB - A pair of diastereomeric peptidomimetics based upon opioid receptor-binding pharmacophore models derived for a series of opioid tetrapeptides was synthesized. Both analogues display high opioid receptor affinity, moderate selectivity for the mu opioid receptor, and are potent, full agonists. PMID- 9873604 TI - Pyrroles and other heterocycles as inhibitors of p38 kinase. AB - Investigation of furans, pyrroles and pyrazolones identified 3-pyridyl-2,5-diaryl pyrroles as potent, orally bioavailable inhibitors of p38 kinase. 3-(4-pyridyl-2 (4-fluoro-phenyl)-5-(4-methylsulfinylphenyl)-pyrrol e (L-167307) reduces secondary paw swelling in the rat adjuvant arthritis model: ID50 = 7.4 mg/kg/b.i.d. PMID- 9873605 TI - Novel protein kinase C inhibitors: alpha-terthiophene derivatives. AB - A series of alpha-terthiophene derivatives were prepared and their protein kinase C inhibitory activity were evaluated. The aldehyde derivatives were most potent inhibitors (IC50 < 1 microM). alpha-Terthiophene monoaldehyde was inactive in the inhibitions of protein kinase A, mitogen activated protein kinase and protein tyrosine kinase. PMID- 9873606 TI - Tropane-based amino acids for peptide structure-function studies: inhibitors of platelet aggregation. AB - Novel tropane (azabicycloheptane) and azabicyclohexane containing amino acids have been prepared and incorporated into analogues of reported inhibitors of platelet aggregation. The influence of these central constraints upon biological activity suggest their utility in peptide structure function studies. PMID- 9873607 TI - The de novo design and synthesis of cyclic urea inhibitors of factor Xa: initial SAR studies. AB - In this report we discuss the design, synthesis, and validation of a novel series of cyclic urea inhibitors of the blood coagulation protein Factor Xa. This work culminates in compound 11, a monoamidine inhibitor of fXa employing a new S4 ligand that reduces the cationic character of these analogs. Compound 11 represents a lead for a series of more potent and selective inhibitors. PMID- 9873608 TI - Design and synthesis of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid, a novel and highly active inhibitor of nitric oxide production in mouse macrophages. AB - New derivatives with electron-withdrawing substituents at the C-2 position of 3 oxoolean-1-en-28-oic acid were synthesized. Among them, 2-cyano-3,12-dioxoolean 1,9-dien-28-oic acid (CDDO) was 400 times more potent than previous compounds we have made as an inhibitor of production of nitric oxide induced by interferon gamma in mouse macrophages (IC50, 0.4 nM). The potency of CDDO was similar to that of dexamethasone, although CDDO does not act through the glucocorticoid receptor. PMID- 9873609 TI - NGB 2904 and NGB 2849: two highly selective dopamine D3 receptor antagonists. AB - N-(4-[4-?2, 3-dichlorophenyl?-1-piperazinyl]butyl)-3-fluorenylcarboxamide and N (4-[4-?2, 3-dichlorophenyl?-1-piperazinyl]butyl)-2-biphenylenylcarboxamide were prepared in several steps from 2,3-dichloroaniline. These compounds were identified as highly selective dopamine D3 receptor antagonists. PMID- 9873610 TI - Studies on the C-terminal of hexapeptide inhibitors of the hepatitis C virus serine protease. AB - Replacement of the C-terminal carboxylic acid functionality of peptide inhibitors of hepatitis C virus (HCV) NS3 protease (complexed with NS4A peptide cofactor) by activated carbonyl groups does not produce any substantial increase in potency. These latter inhibitors also inhibit a variety of other serine and cysteine proteases whereas the carboxylic acids are specific. Norvaline was identified as a chemically stable replacement for the P1 residue of Ac-DDIVPC-OH which was also compatible with activated carbonyl functionalities. PMID- 9873611 TI - Synthesis and biological activity of novel thiazolidinediones. AB - Novel compounds having a dual pharmacophore were synthesised and evaluated for their insulin sensitiser and anti-inflammatory properties in different animal models. PMID- 9873612 TI - Nonsteroidal progesterone receptor antagonists based on a conformationally restricted subseries of 6-aryl-1,2-dihydro-2,2,4-trimethylquinolines. AB - A series of nonsteroidal human progesterone receptor (hPR) antagonists based on conformationally-restricted analogues of a 6-aryl-1,2-dihydro-2,2,4 trimethylquinoline pharmacophore were synthesized and evaluated for their ability to bind to the human progesterone receptor and inhibit progesterone-stimulated reporter gene expression in mammalian cells. PMID- 9873613 TI - 2-Substituted-4-methoxybenzimidazole-based PDE4 inhibitors. AB - A new family of PDE4 inhibitors based on a benzimidazole framework is described. Several of these compounds are orally bioavailable and show efficacy in in vivo models of inflammatory disease. PMID- 9873614 TI - Voltage-dependent ion channel formation by rigid rod-shaped polyols in planar lipid bilayers. AB - In this Letter, we describe the appearance of large, voltage-dependent currents in BLM induced by rigid rod-shaped polyols that function without charge and permanent dipole moment. The capacity of these symmetrical, nonpeptide models to form either short-living nanopores or small ion channels is shown to depend critically on the length of rigid-rod scaffold as well as the nature of the lateral side chains. PMID- 9873615 TI - Conformationally restricted carbamate inhibitors of horse serum butyrylcholinesterase. AB - Conformationally restricted carbamate inhibitors, exo-2-norbornyl-N butylcarbamate (1), endo-2-norbornyl-N-butylcarbamate (2), l-adamantyl-N butylcarbamate (3), and 2-adamantyl-N-butylcarbamate (4) as active site-directed irreversible inhibitors of horse serum butyrylcholinesterase are investigated for values of the dissociation constant (KI), the carbamylation constant (k2), and the bimolecular rate constant (ki). Compound 1 is the most potent inhibitor of the enzyme and the values of KI and ki are 20 nM and 1.1 x 10(5) M-1sec-1, respectively. PMID- 9873616 TI - Functional molecules based on polyazometals. (1). Artificial metalloproteinases prepared by conjugation of polyazometals with poly(allylamine). AB - Polyazometals (PAMs) conjugated with poly(allylamine) manifested high catalytic activity in the hydrolytic cleavage of bovine serum albumin. Based on cleavage sites, the mechanism for the PAM-catalyzed peptide hydrolysis is suggested. PMID- 9873617 TI - Conformationally constrained inhibitors of caspase-1 (interleukin-1 beta converting enzyme) and of the human CED-3 homologue caspase-3 (CPP32, apopain). AB - A systematic study of interleukin-1 beta converting enzyme (ICE, caspase-1) and caspase-3 (CPP32, apopain) inhibitors incorporating a P2-P3 conformationally constrained dipeptide mimetic is reported. Depending on the nature of the P4 substituent, highly selective inhibitors of both Csp-1 or Csp-3 were obtained. PMID- 9873618 TI - Synthesis and cytotoxicity of acetyl-4H,9H-naphtho[2,3-b]thiophene-4,9-diones. AB - Several new acetyl-4H,9H-naphtho[2,3-b]thiophene-4,9-diones were synthesized and evaluated for in vitro cytotoxicity by NCI against seven cancer cell types. 2,7 Diacetyl naphtho[2,3-b]thiophene-4,9-dione (9) showed significant cytotoxicity against leukemia cells with log GI50 values of -7.61 against SR cells and -7.18 against MOLT-4 cells. 3-Acetyl-naphtho[2,3-b]thiophene-4,9-dione (6) also demonstrated potent cytotoxicity in the latter cell line with log GI50 < -8. PMID- 9873619 TI - Reversal of multidrug resistance (MDR) by aspidofractinine-type indole alkaloids. AB - A series of indole alkaloids of the aspidofractinine-type was assessed for their potential in reversing MDR in vincristine-resistant KB cells. Of the compounds tested, kopsiflorine, kopsamine, pleiocarpine, 11-methoxykopsilongine, lahadinine A and N-methoxycarbonyl-11,12-methylenedioxy-delta 16,17-kopsinine were found to show appreciable activity. PMID- 9873620 TI - Synthesis and anticancer evaluation of certain gamma-aryloxymethyl-alpha methylene-gamma-phenyl-gamma-butyrolactones. AB - Certain gamma-aryloxymethyl-alpha-methylene-gamma-phenyl- gamma-butyrolactones were synthesized and evaluated for their anticancer activity. These compounds demonstrated a strong growth inhibitory activity against leukemia cell lines but are relatively inactive against non-small cell lung cancers and CNS cancers. The anticancer potency for aryl portion is in an order of quinoline > 8 hydroxyquinoline > 2-methylquinoline >> naphthalene >> benzene. PMID- 9873621 TI - 2-Pyridinyl-3-(4-methylsulfonyl)phenylpyridines: selective and orally active cyclooxygenase-2 inhibitors. AB - A series of novel 2-pyridinyl-3-(4-methylsulfonyl)phenylpyridines has been synthesized and evaluated with respect to their ability to inhibit the isozymes of cyclooxygenase, COX-1, and COX-2. Optimum COX-2 activity is observed by introduction of a substituent at C5 of the central pyridine. 5- Chloro-3-(4 methylsulfonyl)phenyl-2-(2-methyl-5-pyridinyl)pyridine 33 was identified as the optimum compound in this series. PMID- 9873622 TI - Sialyl Lewis X-polysaccharide conjugates: targeting inflammatory lesions. AB - A novel system for active targeting of inflammatory lesions has been established. A SLeX-CMPul conjugate (2) showed accumulation that was 2.5-fold higher in inflammatory lesions in vivo than a SLN-CMPul conjugate (4) and 300-fold higher than monovalent SLeX (6). PMID- 9873623 TI - Synthesis and immunosuppressant activity of pyrazole carboxamides. AB - A series of novel pyrazole carboxamides is disclosed that demonstrate strong immunosuppressant activity in rodent and human mixed leukocyte response (MLR) assays (IC50 < 1 microM). The synthesis, biological activity, mode of action, and pharmacokinetic properties of this new lead series are discussed. PMID- 9873624 TI - Synthesis and antibacterial activity of 2-alkoxy penems. AB - The phosphite mediated Oxalimide cyclization reaction was extended to 4 dithiocarbonates of N-oxalyl-2-azetidinones to synthesize 2-alkoxy penems 3. In general, the in vitro antibacterial potency of compounds 3 was weak compared to the highly potent 2-alkylthiopenems 2. PMID- 9873626 TI - Exploration of beta-turn scaffolding motifs as components of sialyl Le(X) mimetics and their relevance to P-selectin. AB - Monocyclic and bicyclic lactam units representing beta-turn surrogates were incorporated into a sialyl Le(X) structure by replacement of the natural sugar components. Low micromolar activity was found in a new P-selectin binding assay. PMID- 9873625 TI - Structure-activity studies related to ABT-594, a potent nonopioid analgesic agent: effect of pyridine and azetidine ring substitutions on nicotinic acetylcholine receptor binding affinity and analgesic activity in mice. AB - Analogs of A-98593 (1) and its enantiomer ABT-594 (2) with diverse substituents on the pyridine ring were prepared and tested for affinity to nicotinic acetylcholine receptor binding sites in rat brain and for analgesic activity in the mouse hot plate assay. Numerous types of modifications were consistent with high affinity for [3H]cytisine binding sites. By contrast, only selected modifications resulted in retention of analgesic potency in the same range as 1 and 2. Analogs of 2 with one or two methyl substituents at the 3-position of the azetidine ring also were prepared and found to be substantially less active in both assays. PMID- 9873627 TI - Design and synthesis of dipeptidyl alpha',beta'-epoxy ketones, potent irreversible inhibitors of the cysteine protease cruzain. AB - The dipeptidyl epoxy ketone 4 is a potent, irreversible inhibitor of cruzain, a cysteine protease isolated from Trypanosoma cruzi, with an apparent second order inhibition rate constant of 330,000 sec-1M-1. PMID- 9873628 TI - Design and synthesis of a pyridone-based phosphotyrosine mimetic. AB - A novel pyridone-based tyrosine analog, 6, has been designed to mimic the binding interaction of SH2 domains with phosphotyrosine (pTyr) containing peptides. Synthesis of 6 features a key Pd catalyzed coupling of beta-iodoalanine with phosphonomethyl 4-pyridone triflate. PMID- 9873629 TI - FAD-mediated enzymatic conversion of NAD+ to NADH: application to chiral synthesis of L-lactate. AB - Electroenzymatic reduction of NAD+ to NADH for subsequent use in enzymatic synthesis has been carried out at carbon electrodes bearing lipoamide dehydrogenase (LiDH) immobilized under a Nafion film. The self-mediated electron transfer was made possible by an excess of flavin adenine dinucleotide (FAD) entrapped together with LiDH. Results were compared to those obtained with a similar electrode containing both LiDH and a polymeric form of FAD (pFAD) prepared by anodic polymerization of FAD. PMID- 9873630 TI - Novel cytokine release inhibitors. Part II: Steroids. AB - Steroidal derivatives as IL-1 beta release inhibitors are discussed. PMID- 9873631 TI - In vitro inhibitory effects of DNA topoisomerase II by fernane-type triterpenoids isolated from a Euphorbia genus. AB - Several kinds of naturally occurring fernane-type triterpenoids isolated from a Euphorbia genus were tested on the inhibitory effects of DNA Topoisomerases I (Topo I) and II (Topo II) activities. A-ring cleaved 3,4-seco-8 beta H-ferna 4(23),9(11)-dien-3-oic acid and its 3-hydroxyl derivative were found to be selective inhibitors of Topo II activity without the stabilization of a DNA/Topo II cleavable complex. PMID- 9873632 TI - Synthesis and antibacterial activity of novel 7-(3-substituted-3 or 4 trifluoromethyl-1-pyrrolidinyl)-8-methoxyfluoroquinolones. AB - The titled compounds were synthesized and evaluated for in vitro antibacterial activity. The (3R, 4S)-3-aminomethyl-4-trifluoromethyl derivative (S-34109) was confirmed to be optimal because of its superior activity against quinolone and methicillin-resistant Staphylococcus aureus and low side effect potential. PMID- 9873633 TI - MEK inhibitors: the chemistry and biological activity of U0126, its analogs, and cyclization products. AB - In search of antiinflammatory drugs with a new mechanism of action, U0126 was found to functionally antagonize AP-1 transcriptional activity via noncompetitive inhibition of the dual specificity kinase MEK with an IC50 of 0.07 microM for MEK 1 and 0.06 microM for MEK 2. U0126 can undergo isomerization and cyclization reactions to form a variety of products, both chemically and in vivo, all of which exhibit less affinity for MEK and lower inhibition of AP-1 activity than parent, U0126. PMID- 9873634 TI - A stereoselective alpha-fucosylation reaction using 1-hydroxy 2,3,4-tri-O-benzyl L-fucose donor for the practical synthesis of selectin blocker. AB - A 1-hydroxy 2,3,4-tri-O-benzyl-L-fucose donor affords a high stereoselectivity of glycosylation in the presence of TMSOTf and is a very useful substrate for the preparation of an alpha-L-fucosyl dipeptide in a stereoselective manner. The donor will be a key component in the preparation of an attractively biological selectin blocker 1. PMID- 9873635 TI - 2,3'-disubstituted-2-(2'-carboxycyclopropyl)glycines as potent and selective antagonists of metabotropic glutamate receptors. AB - 2-(9-Xanthylmethyl)-2-(2'-carboxycyclopropyl) glycine 6e is a novel metabotropic glutamate receptor antagonist. A series of alpha, C-3' disubstituted (carboxycyclopropyl)glycines 6f-n were prepared. Antagonist activity was observed for all these compounds at group 2 and group 3 mGluRs. Although they were slightly less active on group 2 mGluRs than non C-3' substituted 6e, the compounds 6f-n were more selective with lesser or no activity on group 1 mGluR subtypes (IC50 values greater than 100 microns). PMID- 9873636 TI - Synthesis and antiproliferative activity of a cyclic analog of dolastatin 10. AB - A cyclic analog of the natural antiproliferative compound dolastatin 10 was synthesized by introducing an ester link between the N- and C-terminal residues which were modified accordingly. The final macrolactonization was performed by using isopropenyl chloroformate and DMAP as reagents. This analog exhibits submicromolar antiproliferative activity against the L1210 and HT29 cell lines and inhibits in vitro tubulin polymerization (IC50, 39 microM). PMID- 9873637 TI - Fused aminotetralins: novel antagonists with high selectivity for the dopamine D3 receptor. AB - Starting from a series of 2-aminotetralins 1, a novel series of N-[4-(4 phenylbenzoylamino)butyl]-octahydrobenzoquinolines and hexahydrobenzoindoles with high potency and selectivity for the dopamine D3 receptor has been designed. The effect of ligand chirality on binding affinity has been established. Selected derivatives (e.g. 2o, 2p) show high functional selectivity and enhanced in vivo properties compared to 1. PMID- 9873638 TI - Structure-based design of peptidomimetic ligands of the Grb2-SH2 domain. AB - We have designed and synthesized a (3-aminomethyl-phenyl)-urea scaffold to mimic the X+1-Asn part of the minimal phosphopeptide sequence, Ac-pTyr-X+1-Asn-NH2, recognized by the Grb2-SH2 domain. The resulting compounds show the same degree of affinity as their peptide counterparts for the Grb2-SH2 domain. This is the first example reported to date of ligands of the Grb2-SH2 domain with substantially reduced peptidic character. PMID- 9873639 TI - Synthesis of benzo[c]quinolizin-3-ones: selective non-steroidal inhibitors of steroid 5 alpha-reductase 1. AB - A short and efficient synthesis of novel benzo[c]quinolizin-3-one derivatives is described. The synthesis is based on the tandem Mannich-Michael cyclization between 2-silyloxy-1,3-butadienes and a N-t-Boc iminium ion. The prepared derivatives are selective inhibitors of human steroid 5 alpha-reductase isoenzyme 1, thus having potential application as drugs for treatment of male pattern baldness and other DHT-dependent skin disorders. PMID- 9873640 TI - Polycyclic aromatic compounds as anticancer agents: synthesis and biological evaluation of some chrysene derivatives. AB - Synthesis and biological evaluation of new chrysene derivatives aimed at the development of anticancer agents were carried out. PMID- 9873641 TI - Diastereoselective synthesis of octahydro-14H-benzo[g]quinolino-[2,3 a]quinolidines. Improved cytotoxic activity against human brain tumor cell lines as a result of the increased rigidity of the molecular backbone. AB - Cis-Octahydro-14H-benzo[g]quinolino[2,3-a]quinolidines 6 were obtained in 6 steps from L-phenylalanine. The key step utilizes a diastereoselective intramolecular EtAlCl2-catalyzed hetero-Diels-Alder reaction. Compounds 6a-f were tested in vitro against human medulloblastoma D283 Med and glioblastoma A-172 and T98G cell lines and showed improved cytotoxicity compared to the corresponding, less rigid pyrido[1,2-b]isoquinolines 1. PMID- 9873643 TI - Quinazolines: combined type 3 and 4 phosphodiesterase inhibitors. AB - A series of quinazolines has been prepared and evaluated for its ability to inhibit cyclic AMP phosphodiesterase type 3, type 4A, 4B and 4D. The most potent inhibitors showed IC50 values in the nanomolar range for type 3 and type 4 isoforms and bind with high affinity to the [3H]rolipram binding site. These quinazolines represent a new family of potent mixed PDE 3/4 inhibitors and are expected to have a therapeutic potential. PMID- 9873642 TI - Beta-analogs of PLG (L-prolyl-L-leucyl-glycinamide): ex-chiral pool syntheses and dopamine D2 receptor modulating effects. AB - Starting from (S)- and (R)-aspartic acid enantiomerically pure beta-proline derivatives were synthesized. These chiral building blocks were transformed into beta-analogs of the dopamine receptor modulating peptide PLG. According to dopamine receptor binding studies, significant enhancement of [3H]pramipexole binding was observed for the isomeres 1a,b and 2a-c. The derivative 1b revealed an activity comparable to PLG. PMID- 9873644 TI - Identification of side chains on 1,2,5-thiadiazole-azacycles optimal for muscarinic m1 receptor activation. AB - Series of analogs to the functional m1 selective agonist, xanomeline (hexyloxy TZTP), were evaluated for their in vitro m1 efficacy in cell lines transfected with the human m1 receptor. Systematic variation of the side chain and the azacyclic ring led to the discovery of potent muscarinic agonists with robust m1 efficacy, all having the phenylpropargyloxy/thio as the side chain. The most selective compound was the phenylpropargylthio-[3.2.1] endo analog 28, which is a potent and efficacious m1 agonist with no m2 activity. PMID- 9873645 TI - Identification of a series of 1,2,3,4-tetrahydroisoquinolinyl-benzamides with potential anticonvulsant activity. AB - A series of N-(tetrahydroisoquinolinyl)-2-methoxybenzamides was identified by high-throughput screening at the novel SB-204269 binding site. SAR studies have provided compounds 4 and 14 with high affinity and good anticonvulsant activity in animal models. PMID- 9873646 TI - 4-Phenyl-4-oxo-butanoic acid derivatives inhibitors of kynurenine 3-hydroxylase. AB - Kynurenine 3-hydroxylase (KYN 3-OHase) is a key enzyme in the kynurenine pathway of tryptophan degradation and its inhibition may be an effective mechanism for counteracting neuronal excitotoxic damage. We present here a new class of inhibitors derived from a structure-activity relationship (SAR) study of the benzoylalanine side-chain of 1. 2-hydroxy-4-(3,4-dichlorophenyl)-4-oxobutanoic acid (8) and 2-benzyl-4-(3,4-dichlorophenyl)-4-oxo-butanoic acid (10) emerged as the most interesting derivatives. Enantiospecific synthesis for both enantiomers of 8 and diastereomeric salt resolution for those of 10 were successfully applied. PMID- 9873647 TI - First synthesis of alternating SATE-phosphotriester/phosphodiester prooligonucleotides on solid support. AB - Prooligonucleotides with alternating S-Acyl-ThioEthyl (SATE) phosphotriester and phosphodiester linkages or a gap were synthesized by elimination of cyanoethyl group with non-nucleophilic base DBU. Their half-lives in the presence of Pig Liver Esterases (PLE) and in total cell extract were determined. PMID- 9873648 TI - Synthesis of 13C-dehydrocoelenterazine and model studies on Symplectoteuthis squid bioluminescence. AB - In the photoprotein of an Okinawan squid bioluminescence of Symplectoteuthis oualaniensis L a dehydrocoelenterazine has been assigned as a chromophoric precursor to its apoprotein. To prove this mechanism, we have established new synthetic route to ca. 100%-13C incorporated dehydrocoelenterazine and coelenterazine at the neighboring carbon of the 2-position of 2,3-dihydroimidazo [1,2a]-pyrazinone skeleton. This 13C enriched dehydrocoelenterazine readily converted in equilibrium between its adduct forms as a diastereomixture with glutathione (GSH) or dithiothreitol (DTT) compounds having sulfhydryl group. Structures of such adducts were fixed under acidic conditions and then discussed by NMR spectroscopy as well as absorbance and fluorescence spectra. PMID- 9873649 TI - PDE4 inhibitors: new xanthine analogues. AB - Novel xanthine analogues are described which are selective PDE4 inhibitors with improved therapeutic potential over theophylline. PMID- 9873650 TI - Pyrrolnitrin and related pyrroles endowed with antibacterial activities against Mycobacterium tuberculosis. AB - During development of nitroheterocycles with potential antimycobacterial activities we have tested against Mycobacterium tuberculosis a number of pyrroles strictly related to pyrrolnitrin, an antifungal antibiotic isolated from Streptomyces pyrrocinia. Some of the tested arylpyrrole derivatives and pyrrolnitrin have shown appreciable inhibiting activity against M. tuberculosis and M. avium. SAR studies well correlate antimycobacterial potency with the presence of halogens in the phenyl ring and a nitro group at position 3 of pyrrole. PMID- 9873651 TI - Hepatoprotective, superoxide scavenging, and antioxidative activities of aromatic constituents from the bark of Betula platyphylla var. japonica. AB - The 50% aqueous methanolic extract from the bark of Betula platyphylla SUKATCHEV var. japonica (MIQ). HARA was found to show potent inhibitory activity on the liver-injury induced by CCl4 or D-galactosamine (D-GalN)/lipopolysaccharide as well as O2- scavenging and antioxidative activities. From the 50% aqueous methanolic extract, two new diarylheptanoids named betulaplatosides Ia (1) and Ib (2) and a new arylbutanoid named betulaplatoside II (3) were isolated together with seventeen known aromatic constituents. 1, 2, and two known diarylheptanoids [(5S)-5-hydroxy-1,7-bis-(4-hydroxyphenyl)-3-heptanone 5-O-beta-D-apiofurano-syl (1-->6)-beta-D-glucopyranoside (6) and aceroside VIII (7)] showed protective activity against D-GalN-induced cytotoxicity in primary cultured rat hepatocytes. Furthermore, several aromatic constituents exhibited potent O2- scavenging and antioxidative activities. PMID- 9873652 TI - Synthesis of anti-rheumatic agent epoxyquinomicin B. AB - Anti-rheumatic agent (+/-)-epoxyquinomicin B was synthesized for a 22% overall yield in eight steps from commercially available 3-hydroxy-4-nitrobenzaldehyde via the intermediate quinone 6 prepared by selective phenol oxidation of 5 by use of Fremy's salt as the key step. PMID- 9873653 TI - Lactate cannot substitute for alanine in d4T-based anti-HIV nucleotide prodrugs- despite efficient esterase-mediated hydrolysis. AB - As part of our on-going effort to deliver masked phosphates of antiviral nucleosides inside living cells we have previously discovered that amino acid derived phosphoramidates are particularly effective. Here we report that lactate analogues, with a simple change of bridging nitrogen for oxygen, are virtually inactive as antiviral agents and apparently do not achieve intracellular nucleoside phosphate delivery. PMID- 9873654 TI - 5,5-trans lactone-containing inhibitors of serine proteases: identification of a novel, acylating thrombin inhibitor. AB - Synthesis of a variety of 5,5-trans fused lactones, related to compounds found in extracts of Lantana camara, has provided a series of novel acylating inhibitors of human thrombin, trypsin, chymotrypsin and human leucocyte elastase. The most effective thrombin inhibitor is 7 with an IC50 of 130 nM and a Kobs/[1] of 4,000 M-1 s-1. PMID- 9873655 TI - Conformationally constrained NO synthase inhibitors: rigid analogs of L-N iminoethylornithine. AB - The synthesis of eight rigid analogs of L-N-iminoethylornithine (L-NIO) is described. The compounds have been evaluated for their inhibition of inducible nitric oxide synthase. Preliminary structure-activity relationships are discussed. PMID- 9873657 TI - Synthesis of C-ring aromatic taxoids and evaluation of their multi-drug resistance reversing activity. AB - The C-aromatic taxoids were synthesized to develop effective inhibitors against drug efflux mediated by p-glycoproteins. Among those tested using multi-drug resistant tumor cells (2780AD), the benzoate 11 exhibited significant activity as potent as verapamil, a well-established MDR reversing agent. PMID- 9873656 TI - Synthesis of benzanilide derivatives as dual acting agents with alpha 1 adrenoceptor antagonistic action and steroid 5-alpha reductase inhibitory activity. AB - Synthesis of benzanilide derivatives which have dual alpha 1-adrenoceptor antagonistic action and steroid 5 alpha-reductase inhibitory activity and their structure-activity relationships is described. PMID- 9873658 TI - Synthesis and evaluation of artificial taxoids with antitumor and multi-drug resistance reversing activities. AB - Artificial taxoids were synthesized and subjected to evaluation of their ability of multi-drug resistance reversing and antitumor activities. While the taxoid 4 could not increase cellular accumulation of vincristine in multi-drug resistant tumor cells, the C4-hydroxy analog 15 showed significant effect. However, these compounds showed weak activities on growth inhibition of cancer cells. PMID- 9873659 TI - Excellent acceleration of the Diels-Alder reaction by microwave irradiation for the synthesis of new fluorine-substituted ligands of NMDA receptor. AB - A series of 6,11-ethanobenzo[b]quinolizinium derivatives was synthesized through the Diels-Alder reaction between azoniaanthracne and the corresponding 1,1 disubstituted olefin. After a systematic investigation for achieving rapid synthesis, it was found that the reaction is accelerated in polar media such as H2O and trifluoroethanol. In particular, excellent acceleration was effected by microwave irradiation. The new fluorine-substituted ligands thus obtained exhibited potential affinity toward NMDA receptors. PMID- 9873660 TI - Preparation of biotinylated allosamidins with strong chitinase inhibitory activities. AB - NaIO4 oxidation of allosamidin (1), a strong inhibitor of family 18 chitinases, followed by a coupling with Biotin Hydrazide afforded its mono- and dibiotinylated derivatives, 4 and 6. Reduction of 4 by NaBH4 afforded its reduced form 5. Each of these three biotinylated derivatives maintained strong chitinase inhibitory activity. Especially, 6 inhibited a Trichoderma chitinase as strongly as 1. PMID- 9873661 TI - Synthesis and in vitro cytotoxicity of 3-substituted-1,8-diazaanthraquinones produced by Lewis-acid catalyzed hetero Diels-Alder reaction. AB - A hetero Diels-Alder reaction of quinoline-5,8-dione with 1-(N,N-dimethylamino)-3 methyl-1-aza-1,3-butadiene proceeded to give 3-methyl-1,8-diazaanthraquinone (100% regioselectivity) in the presence of Lewis-acid catalyst (ZnCl2 or ZnBr2). Subsequent functionalizations of the benzylic methyl group resulted in the 1,8 diazaanthraquinone analogues as potential antitumor agents. The most active compound, 8, exhibited in vitro cytotoxic activity comparable to that of doxorubicin. PMID- 9873662 TI - The synthesis of a new type of anthracene DNA intercalator. AB - A new type of DNA intercalator based on anthracene 3 was synthesized. Preliminary binding studies show high affinity of this probe to CT-DNA. Higher binding constant of this compound (4.0 x 10(4) M-1) as compared with that known for 9 aminomethylanthracene, is caused presumably by enhanced electrostatic interaction. PMID- 9873663 TI - Evaluation of resins for on-bead screening: a study of papain and chymotrypsin specificity using PEGA-bound combinatorial peptide libraries. AB - TentaGel, ArgoGel and PEGA resins were evaluated for on-bead biological screening, using a fluorescently-labelled peptide attached to each and assayed for papain activity. Peptide attached to PEGA was cleaved in near quantitative yield at the expected sites, whilst an identical sequence on TentaGel and ArgoGel beads was hydrolysed in very low yields and nonspecifically on ArgoGel. The compatibility of PEGA with enzymes was further demonstrated by the determination of subsite specificities of papain and chymotrypsin using PEGA-bound peptide libraries, which proved to be similar to those observed in free solution. PMID- 9873664 TI - Rational design of a new series of mixed anti-HIV pronucleotides. AB - MonoSATE aryl phosphotriesters of AZT are able to deliver intracellularly the corresponding 5'-mononucleotide. This process requires activation by an esterase followed by a phosphodiesterase. This finding opens the way to the design of new pronucleotide series. PMID- 9873665 TI - Nitroglycosylated meso-arylporphyrins as photoinhibitors of gram positive bacteria. AB - Novel porphyrins bearing nitro groups and glucosyl moieties were synthesized. The antibacterial activity of these compounds on Escherichia coli and Staphylococcus aureus is described. Results reveal that their photocytotoxicity is markedly dependent on the nature, the number and the linking position of the glycosyl moieties and nitro groups controlling their amphiphilic characters. PMID- 9873666 TI - Triple helix stabilization properties of oligonucleotides containing 8-amino-2' deoxyguanosine. AB - Analysis of the Hoogsteen base pairs of guanine and 8-aminoguanine with cytosine in the gase phase shows a strong stabilization of the 8-aminoguanine.cytosine base pair due to the formation of an extra H-bond. Melting profiles of oligonucleotides carrying 8-amino-2'-deoxyguanosine confirm the triple helix stabilization properties of 8-aminoguanine. PMID- 9873667 TI - Synthesis of novel C7-aryl substituted pyrrolo[2,1-c][1,4]benzodiazepines (PBDs) via pro-N10-Troc protection and Suzuki coupling. AB - Novel C7-aryl pyrrolo[2,1-c][1,4]benzodiazepines (PBDs) have been synthesized via Suzuki coupling between a 7-Iodo N10-Troc-protected PBD carbinolamine and commercially available boronic acids. PMID- 9873668 TI - Design, synthesis and biological activity of a pyrrolo [2,1-c][1,4]benzodiazepine (PBD)-distamycin hybrid. AB - We report the synthesis of a new hybrid 13 which is a combination of the naturally occurring antitumor agent distamycin A 1 and the pyrrolo[2,1 c][1,4]benzodiazepine 11, related to the naturally occurring anthramycin 2. The antitumor activity of the hybrid 13 was tested in vitro and compared to the natural product distamycin 1 and the PBD 11. PMID- 9873669 TI - Oxidative and reductive transformations of epothilone A. AB - The C7 hydroxy group of cytotoxic epothilone A was selectively oxidized using PDC. A selective oxidation of the C3 hydroxy group was accomplished with Me2S/(PhCO2)2 after in situ protection of C7-OH. Reduction of epothilone A or of a C5, C7 dioxo derivative with NaBH4 proceeded at the C5 carbonyl group. Oxidation and hydrogenation of the C16-C17 double bond proved to be difficult but it was easily cleaved with ozone and the resulting keto derivative was transformed to epothilone analogs with different side chains. PMID- 9873670 TI - Derivatization of the C12-C13 functional groups of epothilones A, B and C. AB - Epothilone A reacted with hydrohalic acids to C12-C13 halohydrin regioisomers (ratios: 2:1-4:1), whereas epothilone B gave under the same conditions the isomerically pure C12 halo C13 hydroxy derivative. With non-nucleophilic Bronstedt acids and with Lewis acids a highly solvent dependent product distribution and some unexpected rearrangement products were observed. Epothilone C bearing a double bond between C12 and C13 was regioselectively dihydroxylated or hydrogenated at that position. PMID- 9873671 TI - The cruciferous phytoalexins brassinin and cyclobrassinin are intermediates in the biosynthesis of brassilexin. AB - Following feeding experiments with the tetradeuterated cruciferous phytoalexins brassinin (5b) and cyclobrassinin (6b), leaves of Brassica carinata were elicited with the blackleg causing fungus Phoma lingam and incubated. Spectroscopic and HPLC analyses indicated that both brassinin (5a) and cyclobrassinin (6a) were incorporated into the cruciferous phytoalexin brassilexin (7a). PMID- 9873672 TI - Molecular mechanics parameterization of thionucleoside disulfides for modeling cross-linked duplex DNA. AB - We describe a solution to a molecular mechanics parameterization problem involving disulfide bonds between thionucleosides. Key torsional and bending parameters developed from ab initio calculations were incorporated into the AMBER* force-field in order to accurately represent the disulfide linkage in DNA cross-linked via this bond. PMID- 9873673 TI - Solution-phase combinatorial synthesis of ureas using nitrophenylcarbamates. AB - Nitrophenylcarbamates were reacted with various amines to yield ureas. A high throughput purification of these crude products was achieved by using polymer bound scavengers. PMID- 9873674 TI - Synthesis and hybridization property of an oligonucleotide analog containing a 1',3'-di-O-methylene-alpha-D-fructose backbone. AB - Hydrogen phosphonate monomers of T (thymine) and C(m) (5-methylcytosine) bearing a 1',3'-di-O-methylene-alpha-D-fructose sugar moiety were synthesized and incorporated into an oligonucleotide. Hybridization studies by thermal denaturation experiment indicated that this oligonucleotide did not form a duplex with the complementary RNA target. PMID- 9873675 TI - Orally active indole N-oxide PDE4 inhibitors. AB - This communication describes the synthesis and in vitro and in vivo evaluation of a novel potent series of phosphodiesterase type (IV) (PDE4) inhibitors. Several of the compounds presented possess low nanomolar IC50's for PDE4 inhibition and excellent in vivo activity for inhibition of TNF-alpha levels in LPS challenged mice (mouse endotoxemia model). Emesis studies (dog) and efficacy in a SCW arthritis model for the most potent PDE4 inhibitors are presented. PMID- 9873676 TI - Synthesis of a series of sulfinic acid analogs of GABA and evaluation of their GABAB receptor affinities. AB - A series of gamma-aminobutyric acid (GABA) 1 analogs was prepared in which the carboxylic acid group of GABA was replaced with a sulfinic acid group and their affinity for the GABAB receptor investigated. PMID- 9873677 TI - Electrochemical studies of S-nitrosothiols. AB - S-nitrosothiols (RSNO), such as SNAP, GSNO, and Glc-SNAP-1 exhibited a single, totally irreversible, diffusion controlled reduction peak at potentials of -0.97 V, -0.98 V and -0.91 V, respectively, vs. Ag/AgCl (3 M NaCl) reference electrode. This corresponded to the nitric oxide (NO) release from the S-nitrosothiols. A possible mechanism is proposed for the reduction process. PMID- 9873678 TI - New phthalimide derivatives with potent analgesic activity: II. AB - Seven new phthalimide derivatives (9a-g) with 1,2,4-oxadiazol-5-yl methyl group attached to nitrogen have been synthesized from N-phthaloylglycine 6 and arylamidoximes 7a-g. All of these showed potent analgesic effect with acetic acid induced "writhing" test in mice. One of the better compounds (ID50 = 2.2 mg/Kg i.p.) has been found to be 9a which also demonstrates analgesic activity against inflammatory pain. PMID- 9873679 TI - Molecular structure and biological and pharmacological properties of 3-hydroxy-2 methyl-1-(beta-D-ribofuranosyl or pyranosyl)-4-pyridinone: potential iron overload drugs for oral administration. AB - Replacing alkyl groups by sugar moieties at N-1 position of 3-hydroxy-2-methyl-4 pyridinone did not affect the geometry of the iron chelating sites but increased the hydrophilic nature. The formation of a polymer cluster through the intermolecular hydrogen bonds was also revealed by X-ray crystal structure analysis for the first time in all known 3-hydroxy-4-pyridinone crystal structures. Iron removal from ferritin by the title compounds was more efficient than with DFO. PMID- 9873680 TI - Nonpeptide oxytocin antagonists: potent, orally bioavailable analogs of L-371,257 containing a 1-R-(pyridyl)ethyl ether terminus. AB - Structure-activity studies on the oxytocin antagonist 1 (L-371,257) have identified a new series of high affinity, receptor-selective OT antagonists in which the N-acetyl-4-piperidinyl ether terminus in 1 has been replaced with a 1 (aryl)ethoxy group. PMID- 9873682 TI - Thyrotropin releasing hormone analogs: a building block approach to the construction of tetracyclic peptidomimetics. AB - A building block based approach was used to synthesize a pair of tetracyclic peptidomimetics that constrain all but one of the rotational degrees of freedom of the hypothalamic tripeptide hormone thyroliberin. One of the analogs bound to the thyroliberin endocrine receptor (TRH-R) with an affinity greater than that of an analog without constraints. The tetracyclic peptidomimetics were found to be partial agonists for the TRH-R receptor. PMID- 9873681 TI - 3-Pyridylethanolamines: potent and selective human beta 3 adrenergic receptor agonists. AB - The 3-pyridylethanolamine L-757,793 is a potent beta 3 AR agonist (EC50 6.3 nM, 70% activation) with 1,300- and 500-fold selectivity over binding to the beta 1 and beta 2 ARs, respectively. L-757,793 stimulated lipolysis in rhesus monkeys (ED50 0.2 mg/kg) with a maximum response equivalent to that elicited by isoproterenol. PMID- 9873683 TI - Facile and improved synthesis of 4 beta-aminopodophyllotoxin congeners. AB - An efficient synthesis of 4 beta-aminopodophyllotoxin from 4 beta bromopodophyllotoxin using ammonia, and also a facile synthesis of 4 beta-amino 4'-O-demethylpodophyllotoxin from 4 beta-azidopodophyllotoxin by simultaneous azido reduction and selective demethylation at 4'-position employing chlorotrimethylsilane and sodium iodide, has been described. These are potential precursors for the various 4 beta-amino analogs of podophyllotoxin possessing DNA topoisomerase II inhibition activity. PMID- 9873685 TI - Synthesis of novel fluorinated coumarins: excellent UV-light excitable fluorescent dyes. AB - Two new fluorinated fluorescent dyes, 6,8-difluoro-7-hydroxy-4-methylcoumarin (Marina Blue) and 3-carboxy-6,8-difluoro-7-hydroxycoumarin (Pacific Blue), exhibit excellent photophysical properties among a series of novel fluorinated 7 hydroxycoumarins. Most of these fluorinated coumarins have quantum yields (0.63 to 0.89) equal to or higher than that of the parent compound (0.63), which, in combination with their lower pKaS and higher photostability, make them superior fluorescent dyes for use as reporter molecules in biological systems. PMID- 9873684 TI - Structural determinants of substrates and inhibitors: probing glutamate transporters with 2,4-methanopyrrolidine-2,4-dicarboxylate. AB - Using an intramolecular [2 + 2] photocyclization, 2,4-methanopyrrolidine-2,4 dicarboxylate was prepared as a conformationally locked analogue of glutamate. This compound, in combination with two other pyrrolidine dicarboxylates, has been used to define the structural elements that differentiate substrate and nonsubstrate inhibitors of a high-affinity, sodium-dependent glutamate transporter. PMID- 9873686 TI - Pyrimidinylimidazole inhibitors of CSBP/p38 kinase demonstrating decreased inhibition of hepatic cytochrome P450 enzymes. AB - Pyrimidine analogs of the pyrimidinylimidazole class of CSBP/p38 kinase inhibitors were prepared in an effort to reduce the potent inhibition of hepatic cytochrome P450 observed for the pyridinyl compounds. The substitution of pyrimidin-4-yl, 2-methoxypyrimidin-4-yl, or 2-methylaminopyrimidin-4-yl for pyridin-4-yl effectively dissociates CSBP/p38 kinase from P450 inhibition for this series and furthermore achieves an increase in oral activity. PMID- 9873687 TI - Macrolide biosynthesis: a single cytochrome P450, PicK, is responsible for the hydroxylations that generate methymycin, neomethymycin, and picromycin in Streptomyces venezuelae. AB - The final step in the biosynthesis of methymycin, neomethymycin, and picromycin is an hydroxylation, shown to be carried out by the cytochrome P-450 monooxygenase, PicK. Direct comparison of the relative Kcat/K(m) values for the two substrates, YC-17 and narbomycin, showed a threefold rate preference of picK for narbomycin. PMID- 9873688 TI - Enhancing effects of a mono-bromo substitution at the para position of the phenyl moiety on the metabolism and anti-HIV activity of d4T-phenyl methoxyalaninyl phosphate derivatives. AB - d4T-5'-[p-Bromophenyl methoxyalaninyl phosphate] (d4T-pBPMAP), a novel phenyl phosphate derivative of 2',3'-didehydro-2',3'-dideoxythymidine (d4T) that has an enhanced ability to undergo hydrolysis due to the electron withdrawing properties of its single bromo substituent at the para-position of the phenyl moiety, was found to yield substantially more of the key metabolite alaninyl d4T monophosphate (A-d4T-MP) than the unsubstituted d4T-5'-phenyl methoxyalaninyl phosphate or para-methoxy substituted d4T-5'-phenyl methoxyalaninyl phosphate. d4T-pBPMAP was tested for its anti-HIV-1 activity in peripheral blood mononuclear cells (PBMNC) and thymidine kinase (TK)-deficient CEM T-cells. d4T-pBPMAP was 12.6-fold more potent than the parent compound d4T in inhibiting p24 production (IC50 values: 44 nM vs 556 nM) and 41.3-fold more potent than d4T in inhibiting the reverse transcriptase (RT) activity (IC50 values: 57 nM vs 2355 nM) in HIV-1 infected TK-deficient CEM cells. Similarly, d4T-pBPMAP was more potent than the unsubstituted or para-methoxy substituted phenyl methoxyalaninyl phosphate derivatives of d4T. d4T-pBPMAP did not exhibit any detectable cytotoxicity to PBMNC or CEM cells at concentrations as high as 10,000 nM. Notably, d4T-pBPMAP was capable of inhibiting the replication of a zidovudine (ZDV/AZT)-resistant HIV 1 strain as well as HIV-2 in PBMNC at nanomolar concentrations. To our knowledge, this is the first demonstration that the potency of the d4T-aryl-phosphate derivatives can be substantially enhanced by introducing a single para-bromo substituent in the aryl moiety. PMID- 9873689 TI - Structurally similar small molecule photoaffinity CCK-A agonists and antagonists as novel tools for directly probing 7TM receptors-ligand interactions. AB - Incorporation of photolabile benzoyl (2a-d) or trifluoromethyl-3H-diazirine (3a d) substituents into 1,5-benzodiazepine ligands did not significantly impair the rat CCK-A binding affinity of either agonists or antagonists. The modified agonist ligands also retained functional potency and efficacy in the rat amylase assay. Despite their strong structural similarity, the SAR of this limited set of compounds suggests that these small molecule antagonists and agonists might differ in their mode of binding to the CCK-A receptor. Preliminary affinity results show that representative agonists and antagonists from these series can be used to efficiently covalently label the CCK-A receptor. PMID- 9873690 TI - Design and synthesis of a tetrahydropyran-based inhibitor of mammalian ribonucleotide reductase. AB - A tetrahydropyran-based inhibitor (2) of mammalian ribonucleotide reductase (mRR) has been designed and synthesized based on the heptapeptide, N-AcFTLDADF (1), corresponding to the C-terminus of the R2 subunit of mRR. Inhibition studies revealed that 2 is indeed a competent inhibitor, albeit less potent than 1. PMID- 9873691 TI - Design and synthesis of SH3 domain binding ligands: modifications of the consensus sequence XPpXP. AB - Spirolactam-based Pro-Pro mimetics incorporated in the consensus sequence XPpXP, lead to effective nonpeptide ligands of SH3 domains. PMID- 9873692 TI - Preparation of meta-amidino-N,N-disubstituted anilines as potent inhibitors of coagulation factor Xa. AB - The serine protease factor Xa is a critical enzyme in the blood coagulation cascade. Recently, the inhibition of factor Xa has begun to emerge as an attractive strategy for the discovery of novel antithrombotic agents. Here we describe a series of meta-amidino-N,N-disubstituted anilines as structurally simple and very potent inhibitors of factor Xa. PMID- 9873693 TI - N-substituted octahydro-4a-(3-hydroxyphenyl)-10a-methyl-benzo[g]isoquinolines are opioid receptor pure antagonists. AB - N-Methyl- and N-phenylethyl-(+/-)-1,2,3,4,4a,5,10,10a- octahydro-4a-(3 hydroxyphenyl)-10a-methyl-benzo[g]isoquinolines (4 and 5, respectively) were found to be pure opioid antagonists. These compounds were shown to share many of the characteristics identified with the N-methyl- and N-phenylethyl trans-3,4 dimethyl-4-(3-hydroxyphenyl)piperidine (1 and 2, respectively) including N substituent mediated potency and a lack of N-substituent mediated antagonism. These data suggest that compounds 4 and 5 and the N-substituted trans-3,4 dimethyl-4-(3-hydroxyphenyl)piperidines (1 and 2) may interact with opioid receptors similarly. PMID- 9873694 TI - Syntheses and antitumor activity of cis-restricted combretastatins: 5-membered heterocyclic analogues. AB - A series of cis-restricted combretastatin analogues with 5-membered heterocycles were synthesized and their inhibitory activity against microtubule assembly and cytotoxic activity against the colon 26 adenocarcinoma cancer cell line were evaluated. Some of the heterocyclic analogues showed potent antitubulin activity and cytotoxicity. Compounds 16 and 35 showed marked tumor growth suppression in the colon 26 murine tumor model. PMID- 9873695 TI - Synthesis and biological evaluation of a water soluble phosphate prodrug of 3 aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP). AB - With the aim of improving its biological and pharmaceutical profiles, two water soluble phosphate prodrugs of 3-AP, 3a and 3b were prepared. The detailed synthesis and the preliminary evaluation of these prodrugs are described. PMID- 9873696 TI - Benzimidazole derivatives as arginine mimetics in 1,4-benzodiazepine nonpeptide vitronectin receptor (alpha v beta 3) antagonists. AB - In a 3-oxo-1,4-benzodiazepine-2-acetic acid series of vitronectin receptor (alpha v beta 3) antagonists containing a benzimidazole as a novel arginine mimetic, we examined the effects of benzimidazole modifications and amide substitutions on both activity and pharmacokinetics. PMID- 9873697 TI - Discovery of an imidazopyridine-containing 1,4-benzodiazepine nonpeptide vitronectin receptor (alpha v beta 3) antagonist with efficacy in a restenosis model. AB - In the 3-oxo-1,4-benzodiazepine-2-acetic acid series of vitronectin receptor (alpha v beta 3) antagonists, a compound containing an imidazopyridine arginine mimetic was discovered which had sufficient potency and i.v. pharmacokinetics for demonstration of efficacy in a rat restenosis model. PMID- 9873698 TI - Enantioselective synthesis of a 3'-dephenylcryptophycin synthon. AB - An enantioselective synthesis of tert-butyl (5S,6R)-(E)-5-tert butyldimethylsilyloxy-6-methyl-2,7-octadieno ate, a precursor for the synthesis of the antimitotic macrolides cryptophycin A and arenastatin A (cryptophycin-24), is presented. The key step in the reaction sequence features a crotyl boration that sets both stereocenters that become the C16 hydroxyl and Cl' methyl in the cryptophycins. Homologation of the terminal olefin via a Heck reaction is presented. PMID- 9873699 TI - Synthesis and evaluation of paclitaxel C7 derivatives: solution phase synthesis of combinatorial libraries. AB - A novel and efficient two-step, automated solution phase synthesis of a 26 membered combinatorial chemistry library of paclitaxel C7 esters was accomplished using the HP 7686 Solution Phase Synthesizer. Results of combinatorial synthesis, purification, analysis, and biological evaluation are described. PMID- 9873700 TI - Resveratrol as a new type of DNA-cleaving agent. AB - The first demonstration of DNA cleavage by resveratrol '3,5,4'-trihydroxy-trans stilbene' is presented. Resveratrol mediated relaxation of pBR322 at micromolar concentrations in the presence of Cu2+. Evidence is provided that resveratrol is capable of binding to DNA, and that the Cu(2+)-dependent DNA damage is more likely caused by a copper-peroxide complex rather than by a freely diffusible oxygen species. PMID- 9873701 TI - Potent bicyclic lactam inhibitors of thrombin: Part I: P3 modifications. AB - Peptidomimetic inhibitors of general structure 1 have been prepared. Optimization of the binding affinities of these compounds through variation of the P3 hydrophobic residue is described. Selected substituted bicylic lactams displayed interesting pharmacological profiles both in vitro and in vivo. PMID- 9873702 TI - beta Gal(1-->S-4)beta GlcNac--OR: a galactosidase-stable substrate for alpha(1- >3)fucosyltransferase. AB - 1'-4 Thio-N-acetyllactosamine was chemically synthesized as a galactosidase stable substrate for alpha(1-->3)fucosyltransferase. The product of enzymatic fucose addition was confirmed to be the thio-Le X analog. PMID- 9873703 TI - Evaluation of a structure-based statine cyclic diamino amide encoded combinatorial library against plasmepsin II and cathepsin D. AB - A structure-based 18,900-member combinatorial library was synthesized containing a statine template and three cyclic diamino acids as potential P1, P2-P4 surrogates. Evaluation of this encoded library against two aspartyl proteases, plasmepsin II and cathepsin D, led to the identification of selective inhibitors for each enzyme. PMID- 9873704 TI - Oligonucleotides bearing 5-formyl-2'-O-methyluridine: preference in binding affinity to the NF-kappa B (p50)2 homo- and p50/p65 heterodimers. AB - 5-Formyl-2'-O-methyluridine was incorporated into the various positions of oligonucleotide 26-mers containing the NF-kappa B binding sequence. Some of them showed binding selectivity toward the homo- and heterodimers of subunits of NF kappa B. PMID- 9873705 TI - 10-omega-phenylalkyl-9(10H)-anthracenones as inhibitors of keratinocyte growth with reduced membrane damaging properties. AB - Work aimed at further improving the benefit to risk ratio of the antipsoriatic agent anthralin has led to 10-omega-phenylalkyl-9(10H)-anthracenones, members of which are equally potent as inhibitors of the growth of HaCaT keratinocytes. In contrast to anthralin, induction of membrane injury is strongly reduced as documented by the release of LDH activity from cytoplasm of keratinocytes. PMID- 9873706 TI - Hydroindenic-guanylhydrazones. Synthesis and evaluation as inotropic agents. AB - The synthesis and inotropic activity of two families of hydroindenic compounds are described. Among them, a bis-guanylhydrazone derivative has demonstrated to produce an interesting positive inotropic effect on guinea pig atria, displaying at higher dosis a similar effect to that elicited by digoxin. PMID- 9873707 TI - Fluorescent probes for adenosine receptors: synthesis and biology of N6 dansylaminoalkyl-substituted NECA derivatives. AB - New fluorescent ligands for adenosine receptors are described; these compounds were obtained by the insertion, in the N6 position of NECA (a potent adenosine agonist), of dansylaminoalkyl moieties with alkyl spacers of increasing carbon chain length (from 3 to 12). Among them, the compound with a C6 alkyl spacer proved to be the most interesting one, showing a marked selectivity for the A1 receptor subtype; furthermore, in fluorescence microscopy assays it proved to be able to visualize and localize this receptor subtype at the level of the molecular layer of the rate cerebellar cortex. PMID- 9873709 TI - Ligands for the estrogen receptor, containing cyclopentadienyltricarbonylrhenium units. AB - As model systems for estrogens labeled with technetium-99m that might be used as in vivo imaging agents for estrogen receptor (ER)-positive breast tumors, we have prepared and determined the ER binding affinity of a series of nonsteroidal and steroidal estrogens substituted with a cyclopentadienyltricarbonylrhenium unit. While this organometallic unit interfered with ER binding when it was tethered close to the ligand, those analogs in which it was attached through a 17 alpha ethynyl link (provided that the link was not polar) showed high ER affinity. PMID- 9873708 TI - N-arylrolipram derivatives as potent and selective PDE4 inhibitors. AB - Derivatization of rolipram led to the identification of 3-[4-(3-cyclopentyloxy-4 methoxyphenyl)-2-oxo-pyrrolidin-1-yl]-5-( 3- methoxybenzyloxy)-benzoic acid N',N' dimethylhydrazide (4), a potent and selective inhibitor of PDE4, which inhibits the activation of human leukocytes with pIC50 values in the range of 7.3-7.8, and blocks antigen induced eosinophilia in Brown Norway rats at a dose of 1 mg/kg (i.t.). PMID- 9873710 TI - Synthesis of diacylamines and the preparation of alpha-amino-acylureas, a new type of alpha-amino acid derivatives. AB - Sixteen new and one known unsymmetrical open-chain diacylamines were synthesized by sodium methoxide catalyzed acylation of amides with carboxylic esters and acylamino-carboxylic esters, or acylureas with acylamino-carboxylic esters and alpha-amino acid esters. PMID- 9873711 TI - Synthesis and comparative evaluation of two antiviral agents: beta-L-Fd4C and beta-D-Fd4C. AB - The synthesis of beta-D-Fd4C was achieved in a stereoselective fashion from D xylose. The antiviral activity and cytotoxicity of beta-D-Fd4C was compared with that of beta-L-Fd4C and 3TC (Lamivudine). Of the three agents compared, beta-L Fd4C was found to be the most potent antiviral agent. PMID- 9873712 TI - Aryl ketones as novel replacements for the C-terminal amide bond of succinyl hydroxamate MMP inhibitors. AB - A series of succinyl hydroxamate MMP inhibitors were prepared incorporating an aryl amino ketone moiety in place of the more typical C-terminal amino acid amides. Compounds of the C-terminal ketone series displayed potent inhibition of MMPs. Several compounds of the series were shown to be orally bioavailable. PMID- 9873714 TI - The domino blocks: a simple solution for parallel solid-phase organic synthesis. AB - The domino block is a reaction block for manual and semi-automatic parallel solid phase organic synthesis that simplifies liquid exchange and integrates common synthetic steps. The domino block consists of enclosed reaction vessels, polypropylene syringes, attached to a manifold that clamps the syringes and connects them to a common port. Liquid is removed from the closed reaction vessels by vacuum connected to the common port. The vacuum formed inside each reaction vessel is subsequently used to draw fresh solvent or reagent into each reaction vessel. PMID- 9873713 TI - Growth inhibitory effect of juvenile hormone analogues on epimastigotes of Trypanosoma cruzi. AB - Several compounds, structurally related to the insect growth regulator Fenoxycarb, exhibited interesting inhibition action to control proliferation of Trypanosoma cruzi, the parasite responsible for Chagas' disease. Some of these drugs were shown to be potent growth inhibitors of this parasite. All of these drugs had previously presented juvenoid activity on several non-related bug species such as Tenebrio molitor, Galleria mellonella, Dysdercus cingulatos, and Pyrrhocoris apterus. PMID- 9873715 TI - Thio-sugars. IV: Design and synthesis of S-linked fucoside analogs as a new class of alpha-L-fucosidase inhibitors. AB - alpha-1-Thio-L-fucose derivative 4 and 5 as new alpha-fucosidase inhibitors (K1 = 4.6, and 5.9 microM) have been synthesized in three steps by base catalyzed coupling with bromonitromethane followed by reduction of the nitro group with sodium borohydride/cobalt chloride complex and acetylation. PMID- 9873716 TI - Mixed-backbone oligonucleotides as second-generation antisense agents with reduced phosphorothioate-related side effects. AB - Mixed-backbone oligonucleotides containing alternative phosphorothioate and phosphodiester linkages in the 2'-O-methylribonucleosides segment show increased affinity with complementary targets, increased stability towards nucleases in vitro and in vivo, and reduced phosphorothioate-related prolongation of partial thromboplastin time compared to phosphorothioate oligodeoxynucleotides, thereby providing antisense agents with reduced side effects. PMID- 9873717 TI - Novel phosphate mimetics for the design of non-peptidyl inhibitors of protein tyrosine phosphatases. AB - Benzylic alpha,alpha-difluorosulfonates, alpha,alpha-difluorotetrazoles, and alpha,alpha-difluorocarboxylates of type 5 and 6 were synthesized and examined as potential phosphate biosteres for PTP1B inhibition. The alpha,alpha difluorosulfonates and alpha,alpha-difluorotetrazoles were found to be more effective inhibitors than the analogous compounds bearing the fluoromalonyl group, a phosphate biostere currently being used for PTP inhibition. PMID- 9873718 TI - Restricting the flexibility of crosslinked, interfacial peptide inhibitors of HIV 1 protease. AB - Interfacial peptides of HIV-1 protease were crosslinked with varying length alkyl chains containing either a single cis or trans double bond, or a triple bond to remove degrees of freedom within the tethers. The synthesis of these compounds and their effects on the activity of HIV-1 protease are described. PMID- 9873720 TI - Efficient construction of 6-azasteroids: dual inhibitors of steroidal 5 alpha reductase. AB - A new route to 17 beta-substituted-6-azaandrost-4-en-3-ones, potent dual inhibitors of type 1 and 2 steroidal 5 alpha-reductase, is described. PMID- 9873719 TI - Synthesis of 5-substituted quinazolinone derivatives and their inhibitory activity in vitro. AB - Quinazolinone derivatives I and their methyl esters were synthesized and evaluated as nonclassical lipophilic inhibitors of thymidylate synthase. Compounds Ib and Ic containing OH and CO2H as R substituents, respectively, were most effective, indicating that hydrogen bonding may contribute to the increased inhibitory activity. These compounds further showed high cytotoxic activity against tumor cells in culture. PMID- 9873721 TI - A concise synthesis and in vitro cytotoxicity of new labdane diterpenes. AB - A new series of labdane-related diterpenes have been synthesized from (-) sclareol and assayed in vitro cytotoxicity against mouse and human cancer cells. A key intermediate, homodrimane and furanolabdane derivatives show good in vitro cytotoxicity comparable to those of mitomycin C and adriamycin. PMID- 9873722 TI - Synthesis of 9-membered masked enediyne analogues possessing DNA intercalator and sugar moieties. AB - Syntheses of hybrid DNA cleaving molecules which have the 9-membered enediyne structure of kedarcidin with the DNA intercalator of NCS and/or the sugar moieties were accomplished. Introduction of the sugar moieties to the masked enediyne was achieved via S-alkylation of a bromoacetate derivative with a sugar containing thiol without the need for protection of the sugar moiety. PMID- 9873723 TI - DNA cleaving activities of 9-membered masked enediyne analogues possessing DNA intercalator and sugar moieties. AB - The DNA cleaving properties of various enediyne analogues possessing sugar moieties and DNA-intercalators were investigated. The DNA cleaving experiments show that these hybrids analogues induced sequence-selective DNA cleavage and the simple sugars in the enediyne serve as a DNA recognition element for DNA cleavage. PMID- 9873724 TI - Aryl C-glycosides: physiologically stable glycomimetics of sialyl Lewis X. AB - In the course of the search for physiologically stable, structurally simple, and low molecular weight sLeX mimetics, aryl C-glycosides with carboxylic acid functionality 2 were found to be extremely potent inhibitors against L- and P selectins with IC50 in the low microM range. PMID- 9873725 TI - Potent, non-thiol inhibitors of farnesyltransferase. AB - The structure-activity relationship of a series of non-thiol CaaX analogs, which are inhibitors of farnesyltransferase, is described. These inhibitors contain a substituted phenyl group at the N terminus, which may occupy a novel binding domain on the Ras protein. PMID- 9873726 TI - Anthelmintic beta-hydroxyketoamides (BKAs). AB - We have prepared several anthelmintic coumarins based on the beta hydroxyketoamide (BKA) template and have shown that this template remains valid over a wide range of changes to the coumarin moiety allowing for the inclusion of carbocyclic, bicyclic, and heterocyclic rings. PMID- 9873727 TI - A new series of C3-aza carbocyclic influenza neuraminidase inhibitors: synthesis and inhibitory activity. AB - The synthesis and influenza neuraminidase inhibitory activity of a new series of C3-aza carbocyclic neuraminidase inhibitors are described. Analogues 3c and 3j, bearing a 3-pentyl group, exhibit influenza A inhibitory activities comparable to that of 1. PMID- 9873728 TI - Synthesis of benzocycloalkane derivatives as new conformationally restricted ligands for melatonin receptors. AB - Benzocycloalkane derivatives 1-4 were synthesized as new conformationally restricted melatoninergic ligands. They were prepared by the reaction of the ketones 5 with diethylcyanophosphonate and the reduction of the corresponding cyano compounds or by the Wittig reaction and Curtius degradation to obtain the amines 8. The 1-Cyanobenzocyclobutane derivative was obtained by the benzyne cyclisation reaction. The amines 8 were acylated with acetyl, propionyl or butyryl groups. The affinity of the compounds for chicken brain melatonin receptors was evaluated using 2-[125I]-iodomelatonin as the radioligand. The indanyl (2b,c), tetralin (3a-c) and benzocycloheptane (4c) derivatives were potent compounds with nanomolar affinity and an important enantioselectivity of the receptor was observed with the (+) enantiomers 2b and 3b. PMID- 9873729 TI - Synthesis and morphological reversion activity on srctsNRK cells of pyrimidinylpropanamide antibiotics, sparsomycin, sparoxomycin A1, A2, and their analogues. AB - Three pyrimidinylpropanamide antibiotics sparsomycin (1), sparoxomycins A1, A2 (2, 3), and also six analogues (4-9) have been synthesized by employing asymmetric sulfide oxidation conditions as a key step. Sparsomycin (1) and its alkyl analogues (5-7) showed higher morphological reversion activities on srctsNRK cells than 2 and 3. PMID- 9873730 TI - Potent inhibitors of the MAP kinase p38. AB - The MAP kinase p38 plays a key role in the biosynthesis of the inflammatory cytokines TNF-alpha and IL-1. We have developed a novel series of potent p38 inhibitors that could lead to new methods of treatment for inflammatory diseases such as rheumatoid arthritis and inflammatory bowel disease. PMID- 9873731 TI - Cathepsin B-sensitive dipeptide prodrugs. 1. A model study of structural requirements for efficient release of doxorubicin. AB - A series of lysosomal protease-sensitive peptides attached to doxorubicin (DOX) was prepared as model substrates for internalizing anticancer immunoconjugates and potential antimetastasis prodrugs. Rates of cathepsin B-mediated release of free drug was measured for each, and human plasma stabilities for representative examples. PMID- 9873732 TI - Cathepsin B-sensitive dipeptide prodrugs. 2. Models of anticancer drugs paclitaxel (Taxol), mitomycin C and doxorubicin. AB - Substrates containing doxorubicin (DOX), paclitaxel (taxol), and mitomycin C (MMC) attached to the cathepsin B-sensitive dipeptide Phe-Lys via a self immolative spacer were prepared as model compounds for internalizing anticancer immunoconjugates. Cathepsin B-mediated release rates of free drug, rat liver lysosomal susceptibility and human plasma stability were measured for each. PMID- 9873733 TI - Chemoenzymatic synthesis of iminocyclitol derivatives: a useful library strategy for the development of selective fucosyltransfer enzymes inhibitors. AB - A chemoenzymatic strategy has been developed for the synthesis of libraries of iminocyclitol derivatives for the discovery of new and selective fucosidase inhibitors. PMID- 9873734 TI - Acceptor substrate-based selective inhibition of galactosyltransferases. AB - This paper describes the discovery of glycosyl acceptor analogs as potent and selective inhibitors of alpha-1,3- and beta-1,4-galactosyltransferases. Incorporation of an appropriate aromatic group to the aglycon position of the enzyme's acceptors results in a strong inhibition, representing the first and most potent small uncharged molecules as selective inhibitors of these two enzymes and thus providing a new strategy for the development of selective glycosyltransferase inhibitors. PMID- 9873735 TI - 5-Alkyl 1,2-dihydrochromeno[3,4-f]quinolines: a novel class of nonsteroidal progesterone receptor modulators. AB - A series of nonsteroidal human progesterone receptor (hPR) agonists, 5-alkyl 1,2 dihydrochromeno[3,4-f]quinolines, was synthesized and evaluated in cotransfection and competitive receptor binding assays. The 5-alkyl substitution was shown to be responsible for the agonist activity and substitution at C9 dramatically enhanced the potency. A number of analogues in this series showed activities similar to or better than progesterone in the cotransfection and binding assays and analogue 15 exhibited similar in vivo activity as medroxyprogesterone acetate (MPA) in murine uterine wet weight/mammary gland morphology assays. PMID- 9873736 TI - Novel B-ring modified combretastatin analogues: syntheses and antineoplastic activity. AB - A series of B-ring modified combretastatin analogues were synthesized and their inhibitory activity against microtubule assembly, cytotoxic activity against Colon 26 adenocarcinoma cancer cell line were evaluated. Among these, pyridone derivative (19) showed strong antimitotic activity and cytotoxicity, along with excellent water-solubility. PMID- 9873737 TI - Caged compounds of a 2-deoxyglucose: facile synthesis and their photoreactivity. AB - An o-nitrobenzyl and an o-nitrophenethyl derivatized 2-deoxyglucose (caged 2 deoxyglucoses) were synthesized from 3,4,6-tri-O-acetyl-D-glucal in only two steps in moderate to good yields as isomeric mixtures, which were irradiated at 350 nm to afford a 2-deoxyglucose. Decomposition of the o-nitrophenethyl derivative upon photolysis proceeded more efficiently than that of the o nitrobenzyl derivative. PMID- 9873738 TI - Detection of an anhydride intermediate in the carboxypeptidase A catalyzed hydrolysis of a peptide substrate by solid state NMR spectroscopy and its mechanistic implication. AB - We have detected an anhydride intermediate in the CPA catalyzed proteolytic reaction of Gly-Tyr. It appears that since the zinc-bound water molecule which is believed to attack the scissile amide carbonyl carbon in the hydrolysis reaction is excluded by the N-terminal amino group of Gly-Tyr, the carboxylate of Glu-270 becomes to attack the amide bond to generate the anhydride intermediate. PMID- 9873739 TI - Inhibition of topoisomerase I by naphthoquinone derivatives. AB - Alkannin and shikonin are naturally occurring naphthoquinones. We have tested several derivatives of the title compounds and we have found that naphthoquinones bearing at least one phenolic hydroxyl group are potent inhibitors of topoisomerase I. The ability of the tested compounds to complex Zn++ parallels with a few exceptions their topoisomerase I inhibition properties while their intercalation and redox properties do not. PMID- 9873740 TI - Selective chemical modifications of polymyxin B. AB - Polymyxin B (1) monohydrochloride was converted to the tetra-BOC derivatives 1b and 1c by reaction with di-tert-butyl dicarbonate. The structures of these protected intermediates were established utilizing a degradative sequence that afforded 3 and 5. A method for the deprotection 2,4-dinitrophenylamines to the free amine, utilizing a strongly basic ion-exchange resin, was developed for use in the degradative sequence. The tetra-BOC derivatives 1b and 1c were used to prepare several Polymyxin B derivatives 6-27 at the DAB1 and DAB9-gamma-amine. The antibacterial activity of these selectively functionalized derivatives is reported here. PMID- 9873741 TI - Biologically selective potassium channel openers having 1,1-diethylpropyl group. AB - To find out selective potassium channel openers (PCOs), we synthesized several 3,5-disubstituted phenylcyanoguanidine derivatives and investigated their structure-activity relationships (SAR). As a result, we discovered selective PCOs having 1,1-diethylpropyl group toward antihypertensive activity. PMID- 9873742 TI - Conformationally constrained analogues of diacylglycerol (DAG). 15. The indispensable role of the sn-1 and sn-2 carbonyls in the binding of DAG-lactones to protein kinase C (PK-C). AB - The binding mode of DAG-lactones to PK-C was investigated using the C1b domain from the X-ray structure of the phorbol ester/C1b complex of PK-C delta as a template. Modeling experiments revealed two binding alternatives in which one of the carbonyls of the DAG lactones remained uninvolved with the protein. Experimentally, however, the removal of either sn-1 or sn-2 carbonyls caused a dramatic drop in binding affinity towards PK-C. Although it was not possible to discriminate between the two binding alternatives of the DAG-lactones, the study demonstrates an important role for the additional carbonyl group. The function of this group could be equivalent to that of the C-9(OH)/C-13 (C = O) motif in phorbol esters, which also appears free of interactions in the phorbol ester/C1b complex. This role presumably reflects interaction with the phosholipid head groups required for high affinity binding under the conditions of the biological assays. PMID- 9873744 TI - C24 and C25 substituted marcfortine A derivatives. AB - The dioxepinoindole ring found in marcfortine A (1) is unique among natural products. In order to determine the importance of the substitution pattern of the C24-C25 olefin, we synthesized a variety of analogs at these positions. With the exception of compound 5, none of these compounds exhibited any anthelmintic activity. PMID- 9873743 TI - Potent and selective bicyclic lactam inhibitors of thrombin: Part 2: P1 modifications. AB - The synthesis and antithrombotic activity of a series of nonpeptide bicyclic thrombin inhibitors is described. We have explored the SAR with modifications to the P1 site. The introduction of arginine mimetics at the P1 site led to potent and selective thrombin inhibitors. PMID- 9873745 TI - Synthesis of 4'-epi-iodo-4'-deoxy-daunorubicin, a potential cancer radiotherapeutic agent. AB - We have prepared 4'-epi-iodo-4'-deoxy-daunorubicin (IDDNR)(1), a doxorubicin analog, via a 5-step synthesis involving a protected daunorubicin triflate derivative (4). This triflate derivative will allow the facile and regiospecific nucleophilic preparation of I-125 or Br-80 m labelled analogs of IDDNR. Auger electron-emitting I-125- or Br-80 m-labelled analogs of IDDNR may have potential as cancer radiotherapeutic agents. PMID- 9873746 TI - Design and synthesis of new potent, silent 5-HT1A antagonists by covalent coupling of aminopropanol derivatives with selective serotonin reuptake inhibitors. AB - Hybrid molecules built up by covalent coupling of aminopropanol derivatives (especially pindolol) with antidepressant drugs like fluoxetine, paroxetine or milnacipran were found to be potent and silent 5-HT1A antagonists (KB < 1 nM for 7c and 9a). PMID- 9873747 TI - Synthesis of 2-hydroxy acid from 2-amino acid by Clostridium butyricum. AB - Cultures of Clostridium butyricum type strain in synthetic medium supplemented with various L-2-amino acids revealed the presence of the corresponding 2-hydroxy acid. This metabolite is able to produce the polyester poly(2-hydroxyalkanoic acid). The bioconversion is not stereoselective since D-2-amino acids were also converted. Chiral GC analysis demonstrated that only D-enantiomer is formed from L-leucine. PMID- 9873748 TI - Synthesis and evaluation of a new inhibitor of phosphoglucose isomerases: the enediolate analogue 5-phospho-D-arabinohydroxamate. AB - Designed as a high energy intermediate analogue inhibitor of the potent chemotherapeutic target phosphoglucose isomerases, 5-phospho-D-arabinohydroxamate was efficiently synthesized in a two steps procedure. To date, it proved to be the strongest competitive inhibitor with respect to substrate D-fructose-6 phosphate (Ki down to 98 nM and Km/Ki values up to 513). A comparative inhibition study of this compound and other known strong inhibitors on phosphoglucose isomerases from three different sources is also reported. PMID- 9873749 TI - Orthopaedic surgery in haemophilia--The Tel Hashomer experience: a tribute to Professor Henri Horoszowski. PMID- 9873750 TI - Newer concepts of blood coagulation. AB - In this report we describe an in vitro model of blood coagulation reactions that mimics as closely as possible the in vivo condition. Our model indicates that the tissue factor-factor VIIa complex initiates coagulation by activating small amounts of both factor IX and factor X in the environment of the tissue factor bearing cell. Factor Xa and factor IXa formed in the initial reaction then play very distinct roles in the subsequent interactions of the clotting mechanism leading to a burst of thrombin generation on the platelet surface. Our results also indicate that factor XI can be activated by thrombin in the absence of factor XII and that the function of factor XI is simply to enhance conversion of factor IX to factor IXa resulting in enhanced thrombin generation on the platelet surface. PMID- 9873751 TI - Nonviolent (empathic) communication for health care providers. AB - The purpose of Nonviolent or Empathic Communication Training is to facilitate the flow of information necessary for people to work cooperatively and resolve differences effectively. Such training is widely used in medical communities where the communication with patients and the cooperation between team members are of critical importance for the effectiveness of the treatment. Communication skills are of particular importance for health care providers dealing with patients having chronic diseases such as haemophilia. In addition to the difficulties inherent to the chronicity of the disease, the HIV contamination has dramatically impaired the relationships between patients and health care providers, creating a lot of pain, still alive in both parties. The purpose of this presentation is to offer to health care providers and patients some tools to deal with their feelings and restore effective, compassionate and fulfilling communication. PMID- 9873752 TI - Unsolved problems in haemophilia. AB - Diagnosis and therapy for the vast majority of haemophiliacs in the world remain beyond their reach. Health care costs for hemophilia replacement products keep rising. Global inequities for financing health care result in most hemophiliacs being undertreated. The transfusion-transmitted disease, factor VIII inhibitors, is common. Its predictability escapes detection. Management has progressed substantially and offers many therapeutic modalities. Programs for prophylaxis or immune tolerance induction are impossible for most patients. Thus, the challenge for haemophiliacs is to attain these goals. PMID- 9873754 TI - The molecular basis of haemophilia B. AB - Over the past 15 years, our knowledge of the molecular basis of haemophilia B has increased dramatically. Following the cloning and characterization of the factor IX gene in 1982, major advances have been made in documenting the molecular pathology that underlies this condition. This review will summarize the current state of information in this area, and the reader is referred to the Haemophilia B Mutation Database World Wide Web site at http://www.umds.ac.uk/molgen/haemBdatabase for a complete current listing of the mutations that cause this phenotype. In addition, other recent reviews have discussed complementary issues relating to this topic. PMID- 9873753 TI - The molecular basis of haemophilia A. AB - The cloning and isolation of the human factor VIII (FVIII) gene in the mid-1980s has lead to 10 years of increasing understanding of the genetic and hence the molecular basis of haemophilia A. These studies are not only of enormous potential benefit for accurate carrier detection and prenatal diagnosis in families with haemophilia A, but provide insights into the relationships between genetic defects and their clinical manifestations. These latter studies not only explain and even predict the severity of the disease but may also help towards a better understanding of the basis of inhibitor development. PMID- 9873755 TI - Advances in carrier detection in haemophilia. AB - Increasing worldwide use of molecular genetic analysis is enabling accurate carrier detection for the haemophilias to be made more widely available. Use of DNA polymorphisms in linkage analysis is an accurate method for carrier detection applicable to the majority of families. For those families with severe haemophilia A, the inversion mutation can be sought by most molecular genetics laboratories. For families remaining uninformative by these procedures, a range of point mutation screening techniques is available. Dedicated electrophoresis equipment is enabling use of these techniques to become more widespread. PMID- 9873756 TI - Carrier testing and prenatal diagnosis of haemophilia--utilisation and psychological consequences. AB - Attitudes towards prenatal diagnosis, and abortion vary widely between different countries, religions, cultures and over time. Carrier testing and prenatal diagnosis (PD) of haemophilia have become an integrated part of the comprehensive care for haemophilia in Sweden as well as in many other countries. Almost all carriers are interested in carrier testing if they are aware of the possibility. With the development of PD by chorionic villus sampling in the first trimester, the method became acceptable for many carriers, and it has in Sweden actually had an effect on the incidence of haemophilia in the 1990s. The use of PD is more common among women who perceive haemophilia as a very serious disease and who have a positive attitude towards legal abortion. The main reason for carriers not to use PD was that they do not find haemophilia to be a sufficiently serious disorder to justify an abortion. Women and their spouses are under a great deal of psychological pressure in association with the PD procedure, and the psychological consequences of having to terminate a pregnancy are long-lasting. At follow-up, about 6 years after PD and abortion, these women, however, do not have more signs of psychological distress than women without PD experience. Nevertheless, they must be offered qualified assistance both before and after PD as well as adequate follow-up after an abortion to help them cope with the emotional strain they are under. PMID- 9873757 TI - Can we improve on nature? "Super molecules" of factor VIII. AB - Treatment of haemophilia A requires frequent infusion of plasma- or recombinant derived factor VIII. This regimen is limited due to the high cost and inconvenient access to peripheral veins. In addition, patients frequently develop inhibitory antibodies that limit available therapeutic regimens. Two major advances in factor VIII research over the past 15 years were the ability to isolate homogeneous preparations of factor VIII and the isolation of the factor VIII gene that provided for a detailed biochemical and structural characterization of the factor VIII molecule. With an increased understanding of the requirements for factor VIII function, studies have attempted to produce improved factor VIII molecules for replacement therapy. These findings have produced forms of factor VIII that are more efficiently produced, that are less immunogenic, and that have higher specific activity. The future will see the engineering of novel factor VIII molecules with increased therapeutic efficiency while minimizing inhibitor antibody development. In addition, there are now structural models of factor VIII available that should in the future direct development of novel peptidomimetics that may eventually overcome the requirement for replacement therapy with factor VIII protein. PMID- 9873758 TI - Haemophilia A gene therapy. AB - Gene therapy for haemophilia A would represent a significant improvement over the current treatment by providing prophylactic expression of FVIII and correction of the coagulation defect. Furthermore, a gene therapy protocol allowing simple, infrequent vector administration may extend haemophilia treatment to remote locations world-wide that currently lack access to FVIII replacement therapy. Within the last half decade, significant progress has been made on the development of gene therapy for the treatment of haemophilia A. Recent achievements include high level clotting factor expression in mice, dogs, and monkeys as well as phenotypic correction in haemophiliac mice and dogs. With the efforts that are currently directed toward the improvement of gene transfer vectors and the development of technologies to enable sustained clotting factor expression, gene therapy for haemophilia A will ultimately become a reality. PMID- 9873759 TI - Hepatic gene therapy for haemophilia B. AB - Early retroviral-mediated factor IX gene transfer into deficient dogs showed that constitutive expression of low levels of factor IX which has led to persistent improvement of clinically relevant parameters such as the WBCT and PTT. Conversely, in vivo adenoviral mediated delivery of the factor IX cDNA into hepatocytes of haemophilia B dogs has resulted in greater than wild-type plasma concentrations of clotting factor with complete, albeit transient normalization of haemostasis for a short time. An immune response directed against the vector transduced cells presented a big obstacle to clinical application. However, the future of gene therapy for factor IX deficiency appears bright with the development of fully adenoviral-gene deleted vectors, rAAV and lentiviral vectors which seem to offer safety, therapeutic levels of factor IX and relatively long term persistence. We must proceed with cautious optimism as these vector systems undergo further scrutiny. PMID- 9873760 TI - Safety of coagulation factor concentrates. AB - Many of the adverse effects of the early crude plasma-derived concentrates were ameliorated by increasing their purity. Ironically, this strategy may have increased the risks of inhibitor formation and pathogen transmission due to the addition of processing steps which can alter the immunogenicity of clotting factors and the use of very large plasma pools, as dictated by economic considerations. In the absence of extremely sensitive donor screening, these large pools have a high probability of contamination with pathogens, which may be only partially offset by their removal during protein purification. One approach to minimize the risk of viral transmission is to use recombinant clotting factors produced without the use of human or animal plasma proteins at any step in the manufacturing or formulation process. However, as these proteins are synthesized in mammalian cells, even they pose a theoretical risk of pathogen transmission. For plasma-derived concentrates, the initial viral burden is minimized by screening individual donations and plasma pools with tests which detect virus specific antibodies, protein antigens, or nucleic acid. These techniques are supplemented by non-specific viral reduction steps based on physical partitioning and/or inactivation of pathogens which share chemical or physical characteristics. Prion proteins, the putative causative agents of transmissible spongiform encephalopathies, do not share these characteristics with viruses, and it remains to be determined whether they partition into clotting factor concentrates and whether the current strategies can efficiently remove or inactivate them. For all blood-borne pathogens, active immunization (currently available only for hepatitis B and A) and continued surveillance of susceptible recipients are critical approaches to achieving optimal safety of coagulation factor concentrates. PMID- 9873761 TI - New methods for inactivation of lipid-enveloped and non-enveloped viruses. AB - Two new methods are described for inactivating lipid-enveloped and non-enveloped viruses in plasma-derived products such as coagulation factors and intravenous immunoglobulin (IGIV). Iodine/Sephadex delivers iodine to IGIV solutions in a slow, controlled way and allows for inactivation of > or = 4 logs of porcine parvovirus (PPV), a hardy non-enveloped virus, under conditions which do not measurably damage the structural or functional properties of the IGIV, and with essentially no iodination of the protein. All detectable enveloped and non enveloped viruses were inactivated by this treatment. Gamma irradiation has been successfully used to inactivate viruses at the final vial stage in freeze-dried plasma proteins. Four logs of PPV were inactivated by irradiation in the presence of fibrinogen, factor VIII and alpha 1-proteinase inhibitor (API) at doses of 23, 28 and 30 kiloGray (kGy) respectively, while retaining 93% of fibrinogen solubility, 67% of factor VIII activity and over 80% of API activity. Bovine viral diarrhea virus (BVDV), a lipid-enveloped model for hepatitis C virus, was completely inactivated by radiation doses of 20-30 kGy in these products. Gamma irradiation was less effective in inactivating viruses in freeze-dried IGIV. PMID- 9873762 TI - Prophylactic treatment in Sweden--overtreatment or optimal model? AB - At the haemophilia centre in Malmo, Sweden, regular prophylactic treatment is begun at 1-1 1/2 years of age, before the onset of joint bleeds. The dose and dose interval are optimised by means of pharmacokinetic studies to determine the individual patient's FVIII or IX metabolism, the goal of maintaining a level > 1% of normal being taken as a guideline which experience has shown to yield satisfactory control of bleeding diathesis. An optimal model for prophylactic treatment needs to be applicable to haemophiliacs and acceptable to health authorities in a majority of the countries in the world. To fulfill these criteria, the Swedish model, which has been shown to yield most satisfactory outcome, can hopefully be further refined in the future. Were continuous infusion, using a recombinate concentrate with a prolonged half-life, technically feasible and socially acceptable to the child, we would probably have attained the optimal model of prophylactic treatment. PMID- 9873763 TI - When should prophylactic treatment in patients with haemophilia A and B start?- The German experience. AB - Radiological and orthopaedic outcome in severe and moderate haemophilia A and B patients undergoing long-term prophylactic treatment were prospectively investigated focusing on the age of onset of prophylaxis and the number of joint bleedings prior to treatment. We report on 21 patients with severe and moderate haemophilia A and B receiving prophylactic treatment of between 3.1 and 16.1 years' duration. Three patient groups were evaluated according to the age at onset of prophylaxis. In group I prophylactic treatment was initiated in the first 2 years of life. Patients in group II received prophylaxis at the age of 3 6 years. Late-onset or secondary prophylactic treatment was started at the age of 6 years and above in seven patients (group III). All patients received virus inactivated F VIII or F IX concentrates at dosages of 30-50 IU/kg body weight i.v. three times per week for those with haemophilia A and twice per week for those with haemophilia B. Elbow, knee and ankle joints were investigated at 3-4 yearly intervals according to the radiological and orthopaedic scores recommended by the World Federation of Haemophilia. The total number of joint bleedings before and after start of prophylaxis were recorded in all patients. In group I 7/8 patients had unaffected joints with constant radiological and orthopaedic scores of zero or 1, after a median of 11.25 years of prophylactic treatment. One patient in this group demonstrated mild radiological alterations (score 4). Patients in group II showed neither radiological nor orthopaedic alterations at study entry. Surprisingly, worsening joint scores could be detected despite ongoing prophylaxis after the 3-year interval (median orthopaedic score 4, median radiological score 8). Treatment group III already showed considerable joint damage at study entry with a median radiological score of 11 (0-33) and a median orthopaedic score of 4 (0-11). Despite prophylactic treatment, both radiological (median 19.5, range 2-47) and orthopaedic scores (median 8, range 2-12) deteriorated after 3 years. Prior to onset of prophylaxis, no or only one joint bleeding occurred in treatment group I. In group II, a median of six joint bleeds (range 1-8) was reported before prophylaxis was started. Patients in group III usually experienced a median of more than 10 joint haemorrhages (range 6-10 or more). Under prophylactic treatment the number of joint bleedings decreased significantly in groups II and III. However, radiological and orthopaedic scores increased as a sign of progressing osteoarthropathic alterations in patients reporting more than five joint haemorrhages onset of prophylaxis whereas no joint alterations could be assessed in patients with no or only one joint bleeding episode prior to prophylaxis. Even a small number of joint bleedings seems to cause irreversible osteoarthropathic alterations leading to haemophilic arthropathy. Once apparent, further progression of joint damage could not be arrested despite of prophylactic treatment (groups II and III). In order to prevent haemophilic arthropathy, effective prophylaxis should be started before or at least after the first joint bleeding in severe haemophilia A and B. PMID- 9873764 TI - The use of the Port-A-Cath in children with haemophilia--a review. AB - Port-A-Caths have been used increasingly in children with severe haemophilia. In non-inhibitor patients where Port-a-Caths were used to facilitate long-term prophylaxis, the infection rate is rather low and ranges in the various studies from 0 to 29%, with a median follow-up time of about 27 months. Patients that received the Port-A-Cath for the induction of immune tolerance (inhibitor patients) have a high infection rate of 50% to 83%. Although this percentage is high, good venous access is extremely important, especially in this group. The number of both inhibitor and non-inhibitor patients in the studies are very small, and a prospective survey is important to obtain more adequate data. PMID- 9873765 TI - Haemophilia treatment protocols around the world: towards a consensus. PMID- 9873766 TI - National haemophilia treatment protocols: Canada. PMID- 9873767 TI - Treatment protocol of haemophilia and other congenital bleeding disorders in Italy. Italian Association of Hemophilia Centers (AICE). PMID- 9873768 TI - Guidelines on treatment of haemophilia in Sweden. PMID- 9873769 TI - Haemophilia care within the United Kingdom. PMID- 9873770 TI - Treatment protocols in The Netherlands. PMID- 9873771 TI - Continuous infusion therapy in haemophilia. AB - The application of coagulation factor therapy by continuous infusion (CI) was first suggested by Brinkhous in the early 1950s. The recent introduction of this mode of therapy to everyday practice was made possible after the demonstration of a good stability of most factor concentrates which were also found safe regarding potential bacterial contamination. Other developments included a better understanding of the pharmacokinetics of factors concentrates as well as the availability of a new delivery system. Continuous infusion was shown to be superior to bolus injection (BI) in achieving a stabile haemostatic effect, in the prevention of post-operative bleeding and was found to save between 20-50% in the required factor. This mode of therapy was found effective in haemophilia A and B as well as among patients with inhibitors to FVIII and with von Willebrand disease (vWD). PMID- 9873772 TI - Novel delivery systems for coagulation proteins. AB - Long-term haemophilia prophylaxis with clotting factors administered by alternative delivery modes requires stable liquid formulations of these factors. We developed an aqueous-formulated human coagulation factor IX (hCFIX) with in vitro half-life (T 1/2) of 6 weeks at 37 degrees C and 18 months at 4 degrees C. Upon bolus subcutaneous (s.c.) injection in animals, hCFIX had a bioavailability of up to 16% compared to intravenous (i.v.) dose. When delivered by s.c. implanted pumps, hCFIX attained > 2% of normal human levels in the animal plasma. Hydrogels of hCFIX in a chitosan derivative, N,O-carboxymethyl chitosan (NOCC), released hCFIX slowly in vitro, and when injected s.c., gave prolonged plasma levels over those obtained by bolus i.v. or s.c. injection. Freeze-dried human coagulation factor VIII (hCFVIII) formulated in non-aqueous solvents had in vitro T 1/2 up to 80 days at 37 degrees C. PMID- 9873773 TI - The role of fibrin tissue adhesives in surgery of haemophilia patients. PMID- 9873774 TI - Haemophilia and advanced fibrin sealant technologies. AB - Fibrin sealant, which consists mainly of fibrinogen and thrombin, provides rapid haemostasis as well as tissue sealing and adhesion. Commercial, viral-inactivated products are available in Europe, Canada, and Japan. Liquid fibrin sealant (LFS) has been used clinically in haemophiliacs to perform dental procedures, orthopedic surgeries, non-orthopaedic surgeries, and circumcisions. LFS use is expected to increase as commercial products will soon be available in the US. Recombinant sources and transgenic animal bioreactor systems will replace plasma derived products and become the predominant sources for this product in the next decade. Other areas of innovation include the development of fibrin sealant bandages or dressings, expandable foams, and spray powders which will provide the haemophiliac the ability to rapidly attain control of traumatic haemorrhages prior to hospital treatment with a significant reduction in the use of i.v. clotting factors. Fibrin sealant products have the potential to provide life saving control of haemorrhage, reduction in factor dependency, lower viral exposure risk, and medical care cost reduction. PMID- 9873775 TI - Laser tissue welding: a urological surgeon's perspective. AB - Laser tissue welding has proven its efficacy in the laboratory setting when compared with more traditional modalities of tissue reapproximation. In the clinical environment, several areas including urethral reconstructive surgery have shown great promise. Several technological advancements including solder development, chromophore enhancement and temperature control have improved upon the welding process and have added more precision and reproducibility to the technique. The current potential applications for laser welding in urology are numerous. On a molecular level, growth factor supplementation has certain potential in improving upon weld site healing and wound strength. Laparoscopic surgery with its need for less cumbersome modes of tissue closure is a field that will greatly benefit from the technology of laser tissue welding. Surgical specialties outside of urology are also participating in developing the field of laser welding. In particular, cardiothoracic surgery, otolaryngology, plastic surgery, neurosurgery among others, have utilized the concept of laser tissue welding. There are many ares that have potential use for laser welding that have yet to be explored. Further investigation will likely reveal more applications for this valuable technology. PMID- 9873776 TI - Towards achieving global haemophilia care--World Federation of Hemophilia programmes. AB - The overall purpose of the 'Decade Plan' published in 1992 was to do everything possible to make effective treatment available to people with haemophilia throughout the world. In 1990 it was estimated that there were 350,000 individuals with haemophilia worldwide, with 80% or 280,000 without treatment. By the year 2020 this may have reached the figure 440,000, or the population of Jerusalem. Earlier this century Carroll Birch reported a series of 113 patients where 82 died before their 15th year, and only six survived beyond 40. Thus as Jones has pointed out for the developing world this is a paediatric problem for the WFH to face now. The examples described in this paper illustrating the WFH programmes and the commitment of many people from the 88 member organisations worldwide as well as the doctors and scientists shows what can be done through cooperation. There is no doubt that these achievements are capitalising from the unique quality of WFH from the beginning, that is a membership which includes both those with haemophilia and their families, and those with an interest in treating haemophilia. PMID- 9873777 TI - Management of haemophilia in the developing world. AB - The problems with management of haemophilia in developing countries are poor awareness, inadequate diagnostic facilities and scarce factor concentrates for therapy. The priorities in establishing services for haemophilia include training care providers, setting up care centres, initiating a registry, educating affected people and their families about the condition, providing low-cost factor concentrates, improving social awareness and developing a comprehensive care team. A coagulation laboratory capable of reliably performing clotting times with correction studies using normal pooled, FVIII and FIX deficient patient plasma and factor assay is most essential for diagnosis. More advanced centralized laboratories are also needed. Molecular biology techniques for mutation detection and gene tracking should be established in each country for accurate carrier detection and antenatal diagnosis. Different models of haemophilia care exists. In India, there is no support from the government. Services, including import of factor concentrates, are organized by the Haemophilia Federation of India, with support from other institutions. Haemophilia is managed with minimal replacement therapy (about 2000 i.u./PWH/year). In Malaysia, where the system is fully supported by the government, facilities are available at all public hospitals and moderate levels of factor concentrates are available 'on-demand' (about 11,000 i.u./PWH/year) at the hospitals. Haemophilia care in South Africa is provided through major public hospitals. Intermediate purity factor concentrates are locally produced (about 12,000 i.u./PWH/year) at low cost. The combined experience in the developing world in providing haemophilia services should be used to define standards for care and set achievable goals. PMID- 9873778 TI - Factor concentrates for haemophilia in the developing world. AB - The success in the management of haemophilia in the last two decades has been predominantly due to the availability of sufficient quantities of safe factor concentrates. Unfortunately, the prohibitive cost of these products has prevented this benefit from being available to the vast majority (approximately 80%) of haemophiliacs living in the developing world. A few developing countries have established facilities for the production of low- to intermediate-purity factor concentrates locally. The infrastructure required to achieve this can be very basic. The experience in South Africa, Thailand, Cuba and Brazil, described herein, shows that this approach provides factor concentrates which are very economical in comparison with more purified commercial products. This has had a major impact on the quality of haemophilia care in these countries. Wide availability of low-cost factor concentrates has made these products accessible to a large number of haemophiliacs and even made home therapy possible. The effort to provide these products results in improvement of the blood transfusion services. This, in turn, contributes to better facilities for patients with other transfusion-dependent diseases and society in general. Installation of small plasma fractionation plants is also a viable option. This not only allows processing of large pools of plasma for greater quantities of factor concentrates but also provides albumin and immunoglobulin. The revenue generated from the sale of the other products has been used to improve and subsidize haemophilia care. It is concluded that local production of intermediate purity factor concentrates in developing countries is absolutely necessary. A well organized transfusion service is required to collect adequate quantities of plasma for fractionation. PMID- 9873779 TI - Rehabilitation in haemophilia--options in the developing world. AB - Musculoskeletal care is provided in many different settings by many different providers. Physicians are better prepared to treat acute diseases, acute manifestations and have problems in following chronic patients and to preventing losses of functional independence. Haematoma affecting muscles and joints is the most common musculoskeletal finding in haemophilic patients. Proper treatment is fundamental to prevent disability. Treatment aims at restoring the original muscle trophism and strength because joint stability is dependent on the uniform distribution of muscle power over different joint surfaces. Physical rehabilitation continues until complete anatomical and functional recovery is achieved. Coordination and proprioceptive stimulation complement the exercises to recover original muscle strength. Recurrent haemorrhages usually progress to chronic synovitis, the leading cause of motor disability and joint degeneration in haemophilic patients. Moreover, chronic synovitis is a predisposing factor for repetitive episodes of haemarthrosis, a leading cause of joint deformities and severe muscle atrophy. At this point, an efficient treatment alternative should be put into effect to restrain bleeding and prevent synovitis-related joint degeneration, which is synovectomy. Synovectomy is an effective alternative, less costly and easier to perform. Rifampicin synovectomy is advantageous because it is safe, not aggressive to the patient, can be repeated if necessary, and it does not prevent the use of either radiotherapy or surgery after, if necessary. Once the sequelae have set in, treatment tends to be conservative, and it aims at giving the patient functional independence for gait and other activities. The most common limb deformities seen in these patients are knee and elbow flexion, but the patient should also be assessed for compensatory scoliosis, hyperlordosis and lower limb length discrepancies. Chronic joint pain is a rather common complaint in this patient population with early arthrosis, and special drugs and physical treatment must be put into action. Rehabilitation is often inexpensive and very efficient. PMID- 9873780 TI - State-of-the-art principles and practices of medical economics. AB - In an era of increasing constraints in health care expenditure, it has become necessary for the practising physician to utilise resources economically. Due to expenditure ceilings in many countries, the rational allocation of health care resources has become a central goal in health policy. Medical economics is the discipline which covers the issues of economic evaluation, policy and decision analysis and outcomes research. This paper discusses the fundamentals of this new type of research, highlighting the importance of such results in formulating health policy. Undoubtedly, medical economics will play a similar role as evidence-based medicine and clinical epidemiology in the continuous education within the medical community. This holds specially true for costly indications such as haemophilia and other coagulation disorders. PMID- 9873781 TI - Resource utilisation in haemophiliacs treated in Europe: results from the European Study on Socioeconomic Aspects of Haemophilia Care. The European Socioeconomic Study Group. AB - The objectives of our study was to answer the following questions: (1) What are the socioeconomic costs and benefits of different haemophilia treatment strategies? (2) Which savings can be achieved through a comprehensive care program? (3) How are haemophiliacs integrated into their social life? (4) What are the annual costs of treating haemophilia from a third party payer perspective? In this cross-sectional study, 840 consecutive haemophiliacs attending 16 haemophiliac treatment centres in 10 European countries at were entered in the study. The following six parameters were analysed: sociodemographic patient data, the type of substitution (on demand or prophylaxis), transfusion-transmitted diseases, the functional joint status, quality of life as well as the total costs of therapy. Three kinds of substitution modalities were distinguished: on demand therapy, pure and modified prophylaxis. The average number of medical visits, the average length of hospital stay, regular drug therapy and the average number of days off work per bleeding joint were recorded from the medical charts as a basis for the subsequent assessment of the costs of therapy. The total study sample comprised of 566 patients. Interestingly, statistically more significant (P = 0.0005) patients were unemployed in the on-demand group (14%) than in the prophylaxis group (3.4%). There was a higher ratio of white collar workers in the on-demand group as compared to the prophylaxis group and vice versa for blue collar workers. The distribution of blue collar workers, clerk, self-employed persons, students and housemen were quite comparable between the patients of the on-demand and prophylactic therapeutic regimen. The labor force participation rate was 55.2% in the patients in the on demand and 59.3% in the patients in the modified prophylactic group. The labor force participation rate across all patients was 56%. Patients on demand therapy used on average 38.3 (median: 12.5) units/kg/week as compared to 68.6 (median: 6) units/kg/week in the prophylaxis group. The mean usage across all patients was 46.1 (median 19.7) units/kg/week. There were an average of 7.4 (median: 3) joint bleeds per patient across all patients during the 6-month observation period. In the on-demand group 8.8 (median: 5) joint bleeds were recorded versus 3.1 joint bleeds (median: 0) in the prophylactic group. This difference was statistically different (95% CI of difference: 4.33 7.07). Our data suggest that patients receiving prophylactic clotting factor therapy require less additional health care resources, mainly due to the reduction in the number of joint bleeds. PMID- 9873782 TI - Synovium in haemophilic arthropathy. AB - Synovium is an essential component of the joint and plays a critical role in maintaining a balance between physiological processes and pathological changes in the joint. Recurrent intra-articular bleeding as occur in haemophilia induce pathological synovial changes in the joint. From a certain point on, synovitis inevitably plays a major role in joint destruction, although in the early phase of haemophilic arthropathy its role may be secondary to cartilage damage as a result of the direct effects of blood on cartilage. The changed haemosiderotic, synovial tissue produces catabolic cytokines and enzymes harmful for cartilage. PMID- 9873783 TI - The destructive capabilities of the synovium in the haemophilic joint. AB - The pathogenesis of chronic haemophilic synovitis and the destructive capabilities of the synovium are multifactorial. As a result of repeated haemarthroses, synovial hyperthrophy occurs, iron and cellular debris are phagocytized, and hydrolytic enzymes synthesized by the proliferating synovium are released into the joint. An inflammatory process develops, and the nutrition of the articular cartilage is affected by the abnormal synovial fluid, immobilization of the joint, and the presence of fibrous adhesions. Chronic hypertrophic synovitis is a progressive destructive condition in the joint and, unless it is treated, the joint is doomed. With the development of cartilaginous destruction, progressive chondrolysis and eventual osteoarthritis set in. PMID- 9873784 TI - Control of the synovium in haemophilia. AB - Adequate factor substitution is a prerequisite for successful conservative or operative treatment of haemophilic synovitis. Subsequent prophylactic factor substitution can prevent further joint problems in the future. Orthopaedic treatment regimes are dependent on the classification of the synovitis and the stage of the arthropathy. Synovitis can be classified into acute and chronic forms. The arthropathy is classified according to Arnold and Hilgartner. When the synovitis has already become chronic, early arthroscopic synovectomy is recommended in order to prevent the otherwise inexorable progress of the arthropathy. Failure to treat in the early phase of the pathology causes problems in the correction of further stages. The progression of the joint disease can be shown both clinically and radiologically as the arthropathy develops, and influences the treatment. PMID- 9873785 TI - Haemophilic synovitis: is rifampicin an alternative? AB - Rifampicin synoviorthesis has been empirically used for the treatment of haemophilic synovitis for some time. This paper reports on the experience of three Latin American centers with this treatment and compares it with radioactive synoviorthesis results. Chemical synoviorthesis with rifampicin is best indicated in younger patients (< 15 years) and small joint (ankles and elbows). PMID- 9873786 TI - Treatment of acute and chronic synovitis by non-surgical means. AB - This paper reviews personal experience in the treatment of recurrent haemarthrosis and chronic synovitis by non-surgical means. Experience with synoviorthesis with rifampicine and radioactive colloids is analyzed, and a multiple chromosomal study to demonstrate safety of radioactive injections is described. The results obtained are so very satisfactory as to recommend non aggressive synoviorthesis as the treatment of choice to prevent recurrence of bleeding. Long experience in the treatment of chronic arthropathy with intrarticular corticosteroids and hyaluronic acid has shown very promising results. PMID- 9873787 TI - Functional kinesiology in haemophilia, an area yet to be explored. AB - In haemophilic arthropathy there is a progressive limitation of the range of motion (ROM) which may lead to disabilities in the activities of daily living (ADL). In the literature the pathology of haemophilic arthropathy is described extensively, but only one paper describing functional limitations caused by limited range of motion (LOM) in haemophilia was found. The aim of the pilot study was to estimate on theoretical grounds, how many patients with haemophilia might suffer from functional disabilities. MATERIAL: ROM of elbows, knees and ankles of 155 Haemophilia A and B patients. METHODS: Flexion and extension were measured with an ordinary goniometer. The ROM of joints of patients with haemophilia was compared with normal values. RESULTS: 39 of 155 patients had a normal ROM in both elbows; 22 in one elbow; 34 patients had disabilities in ADL with both arms; 14 with one arm; 18 were able to compensate; 89 had no problems; 79 of 155 patients had a near normal ROM of both knees; 38 patients could not ride an ordinary bicycle. CONCLUSION: Only limited data are available concerning the normal ROM needed for individual ADL. Until additional data are available, it is not possible to predict which patients will be disabled in their activities of daily living and individual counselling should be done during the yearly outpatient comprehensive care clinics. Conservative and surgical measures should be taken to ensure elbow flexion of at least 120 degrees and knee flexion of 100 degrees for Western societies. In Asian countries patients with haemophilia need maximum knee flexion and ankle dorsi flexion. PMID- 9873788 TI - Proprioceptive training in haemophilia. AB - Joint and muscle injury associated with direct damage to the tissues and muscle atrophy may ensue following immobility. Rehabilitation of the injury is linked with the return to normal functional values such as range of motion, muscle strength, and muscle tone. It is, however, likely that subtle changes or differences still exist in the site of injury or haemarthrosis. In particular proprioception may be distorted due to the direct injury of sensory receptors and to feedback systems. The implications of such damage are important, where proprioception plays an important part in the control, timing and organisation of coordinated bodily actions. PMID- 9873789 TI - Muscle rehabilitation in haemophilia. AB - Musculoskeletal dysfunction is a common manifestation of haemophilia. This dysfunction may be associated with imbalances between muscle groups. Evidence emerging from the literature suggests that the rehabilitation of this dysfunction is very relevant for the patient with musculoskeletal problems. Treatment of muscle imbalance may be linked with a reduction in recurrence of symptoms. Further research is needed to establish the relevance of this area in patients with haemophilia but the clinical evidence supports the developing work in this field. PMID- 9873790 TI - How do some haemophiliacs develop inhibitors? AB - The purpose of this presentation is to concentrate on three issues: Is there an explanation for the large difference in reported incidence rates of inhibitor formation in haemophiliacs? Why do not all severe patients develop inhibitors? And how on the contrary do some patients with mild haemophilia develop inhibitors? PMID- 9873791 TI - The genetic basis of inhibitor development in haemophilia A. AB - Inhibitor development is now the main complication of replacement therapy in haemophilia A. Given that most severely affected patients make no detectable factor VIII, it is perhaps surprising that only approximately 30% actually mount an immune response to factor VIII as a foreign antigen. Those that do mostly have major factor VIII gene lesions. The association of HLA genotype with inhibitors in patients with identical mutations is weak. Environmental factors may be more important than genetic in antibody response to factor VIII. PMID- 9873792 TI - The natural history of the immune response to exogenous factor VIII in severe haemophilia A. AB - The development of inhibitory antibodies to factor VIII (fVIII) in severe haemophilia A patients is a serious therapeutic complication. Using a highly sensitive immunoprecipitation (IP) assay which measures all anti-fVIII antibodies, we have tested severe haemophilic plasmas from two clinical studies. Inhibitor titres in the range of 0.4 to 1 Bethesda units/ml (BU/ml) could not be verified by IP as being due to an immune response to fVIII in 35% of plasmas tested. Low fVIII recoveries were likewise correlated with the presence of antibodies in 29% of plasmas tested. However, 16% of plasmas without inhibitor titres had immune responses as measured by IP. The rapidity of antibody appearance did not allow their effective detection by IP before development of inhibitor titres. These results suggest that the IP assay can provide a valuable confirmation of anti-fVIII antibody production when the Bethesda assay is low or negative and where clinical observations suggest their presence, but they cannot be used reliably to detect early immune responses. PMID- 9873793 TI - Factor VIII immunogenicity. AB - The immunogenicity of factor VIII depends on the interaction of multiple parametres including host susceptibility and characteristics of the factor VIII preparations. We briefly review here the basic mechanisms by which tolerance to self is established, maintained, and possibly broken in the context of haemophilia A, with special emphasis on the severity of the disease. Reference is also made to the situation observed in healthy individuals and patients with autoantibodies to factor VIII. PMID- 9873794 TI - Factor VIII inhibitors in mild and moderate-severity haemophilia A. AB - Inhibitors are an uncommon complication of mild haemophilia, occurring in 3-13% of patients and usually arising during adulthood. The risk of inhibitor development in this group appears to be associated with relatively few high-risk factor VIII genotypes clustered in the A2 and C2 domains, especially the Arg593 Cys and the Trp2229-Cys mutations. Kindreds with these mutations have an inhibitor incidence of up to 40%. These mutations may induce a stable conformational change in the factor VIII molecule rendering it antigenically distinct from wild-type factor VIII. Inhibitors in mild haemophilia usually cross react with endogenous factor VIII reducing the basal VIIIC to < 0.01 IU/ml, and causing spontaneous bleeding. This bleeding is sometimes severe and life threatening, two-thirds of patients developing a pattern of soft tissue, gastrointestinal (GI) and urinogenital bleeding reminiscent of acquired haemophilia. Bleeding has been treated with human and porcine factor VIII, bypass therapy and DDAVP. Recombinant factor VIIa and DDAVP have the advantage that they do not induce an anamnestic rise in inhibitor titre. About 60% of these inhibitors disappear in the remainder over a median of 9 months. Few of these inhibitors recur, suggesting that most such patients have become tolerant. The inhibitors persist long-term and remain troublesome in about 40% of patients. The limited data available on immune tolerance induction in this group indicate a generally poor response to this approach. Two of nine achieved tolerance, with a partial response in a further four. Inhibitors are an uncommon but life threatening complication of haemophilia. This complication should be considered when selecting the treatment modality for patients with a family history of inhibitors, and DDAVP used whenever possible. PMID- 9873795 TI - Safety, efficacy and lessons from continuous infusion with rFVIIa. rFVIIa-CI Group. AB - Continuous infusion with coagulation factor concentrates has only been widely adopted during the last decade and with recombinant activated factor VII (rFVIIa) only during the last 2 years. Still the accumulated number of days on continuous infusion with this product amounts by now to more than one year, 35 patients and 26 surgical procedures. The experience is reviewed here and compared with data from the conventional bolus dose regimen. PMID- 9873796 TI - Immune tolerance: a synopsis of the international experience. AB - Because of the increased morbidity and cost of care associated with inhibitor development, immune tolerance therapy (ITT) is of crucial value in the care of haemophilia. The 24-year experience with this modality, primarily in the treatment of factor VIII inhibitors, has included the use of both high and low doses of clotting factor, with and without immune modulation. Overall success rates for ITT in haemophilia A have been similar (63-83%), while median time to IT has been variable (1.2-24 months). The role of type and purity of clotting factor used remains unclear. Three immune tolerance registries have suggested the potential importance of treatment parameters such as pre-induction inhibitor titer and daily factor dose in the prediction of successful outcome. Ultimately, prospective randomized studies of ITT are required to definitively compare therapeutic regimens with respect to efficacy, safety, and cost effectiveness. PMID- 9873797 TI - Management of haemophilia B patients with inhibitors and anaphylaxis. AB - The development of inhibitor antibodies is a serious complication of haemophilia in young children. Occurrence of anaphylaxis at the time of inhibitor development is a recently described complication unique to haemophilia B. Management of these inhibitor patients with allergy is complicated due to the absence of any readily available products for treatment of acute bleeding episodes. Clinical experience suggests that recombinant activated factor VII is the most appropriate and logical treatment for acute bleeding episodes in these patients. From the limited information available regarding immune tolerance induction (ITI) in these patients, it appears that ITI regimens have been only minimally successful and are associated with a high rate of complication (nephrotic syndrome). PMID- 9873799 TI - Vaccines for prevention of viral hepatitis. AB - At the end of the 20th century active immunization against hepatitis A and B has been introduced into routine medical practice. Therefore, patients at risk, and especially those with coagulation disorders, should be immunized at the earliest age possible. In contrast, development of an efficacious vaccine against hepatitis C remains an important goal for the next decade. PMID- 9873798 TI - Therapy for chronic hepatitis B and C infection in haemophilia. AB - Haemophiliacs, until recently, have been at risk of hepatitis B and C infection. Substantial numbers of patients remain persistently infected. Several lines of evidence suggest that these diseases will cause considerable morbidity unless therapy can successfully reduce viraemia and prevent disease progression. Cessation of viral replication may prevent progression to hepatic fibrosis. PMID- 9873800 TI - Assessment and treatment of liver disease in Japanese haemophilia patients. AB - We studied the prevalence of the hepatitis C virus (HCV), human immunodeficiency virus (HIV) and GB virus C or hepatitis G virus (GBV-C/HGV), and characteristics of infections in Japanese haemophilia patients. Haemophilia patients were highly infected with HCV (88.2%) because of frequent use of unheated blood concentrates. Analysis for HCV genotypes revealed characteristics of HCV infection in haemophilia patients. Japanese haemophilia patients were highly infected with rare genotypes in Japan: genotype 1a (26.5%), genotype 3 (14.5%) and genotype 4 (2.4%). HIV infection was observed in 32.3% of haemophilia patients. HCV quasispecies (clones) and direct sequencing were investigated in patients with a single HCV genotype in the hypervariable region 1 of HCV, which resulted in a high degree of diversity. This indicates that even a single genotype of HCV might have multiple origins. GBV-C/HGV infection was noted in 20.9% of Japanese haemophilia patients. Over 40 haemophilia patients with chronic hepatitis C have been treated with interferon alpha for 6 months at total doses of 480-720 million units. About 38% showed clearance of HCV RNA from serum. Six patients with HIV infection were included in the study and they did not show eradication of HCV from the serum. This might derive from that they had high serum HCV RNA titers and genotype 1a or 1b. Histologic assessment was performed in 36 haemophilia patients with HCV. No case showed a histologically normal liver. Hepatic fibrosis in the biopsy specimens was classified into five stages of fibrosis and compared with serum hepatic fibrosis markers. Serum hyaluronic acid mostly correlated with hepatic fibrosis (r = 0.78, P < 0.0001) followed by type IV collagen (r = 0.38, P < 0.05). This suggests that estimation of serum fibrosis markers might be substituted for liver biopsy in haemophilia patients. PMID- 9873801 TI - Progression of HIV in haemophilia. AB - The recent elucidation of the life cycle and dynamics of the human immunodeficiency virus (HIV) and technological advances in development of the HIV RNA PCR assay for sensitive detection of viral load have revolutionized the diagnosis, management, and treatment of HIV infection. Beginning with initial infection, there is unremitting, high-level viral replication that persists throughout the course of HIV infection. The measure of the amount of virus present in plasma, HIV viral load, is the single most important predictor of HIV progression, the best indicator of immune system decline, and the best guide for initiating and monitoring antiviral treatment. Further, HIV viral load has become the new yardstick against which other markers, including CD4 number, age, chemokine receptor mutations, cytotoxic T-cell responses, and neutralizing antibody titers are assessed. For individuals with haemophilia, additional 'markers' may have significant impact on the outcome of HIV disease. Chronic factor concentrate treatment has led to transfusion-associated hepatitis, co infection with hepatitis C (HCV), and chronic liver disease. The latter may become accelerated with HIV progression and may lead to hepatotoxicity with antiviral drug therapy. Chronic factor concentrate treatment has also been associated with immunosuppression, including both B- and T-cell immune defects. In HIV(+) haemophilic men, this immune deficit has led to lower CD4 counts with HIV progression and poorer CD4 response to antiviral drugs than in gay men. The underlying haemophilic bleeding tendency may result in significant haemorrhage with HIV-associated immune thrombocytopenia and with protease inhibitor antiretroviral therapy. Although AIDS is the leading cause of death in this group, the reduction in the size of the haemophilia population over the next two centuries is estimated to be small, and survival should improve as better antiviral therapeutics are identified. PMID- 9873802 TI - Resistance to HIV protease inhibitors. AB - Resistance to the HIV-1 protease inhibitor indinavir involves the accumulation of multiple amino acid substitutions in the viral protease. A minimum of 11 amino acid positions have been identified as potential contributors to phenotypic resistance. Three or more amino acid substitutions in the protease are required before resistance becomes measurable (> or = four-fold). Further losses in susceptibility follow the stepwise accumulation of additional amino acid substitutions, indicating that antiviral activity (selective pressure) is maintained despite the appearance of multiple amino acid substitutions in the viral protease. Importantly, the sequential nature of these changes indicates that the effects of these substitutions are additive, and that the evolution of resistance is driven by viral replication. This result has significant implications for therapy. It predicts that viral variants resistant to indinavir are unlikely to pre-exist in protease inhibitor-naive patients, and further, that high-level resistance can only develop if the virus is allowed to replicate in the presence of the drug. The use of indinavir in combination with other antiretroviral agents has been demonstrated to dramatically reduce the incidence of resistance mutations, suggesting that with maximal suppression of viral replication, long-term control of HIV-1 infection may be achievable. Thus, the goal of therapy must be to never to allow the virus to replicate. This can be best accomplished by initiating therapy with a maximally suppressive regimen, to reduce viral replication as much as possible, and by imposing a high genetic barrier to resistance. Previous use of other protease inhibitors or inadequate adherence to therapy may compromise the long-term benefit of indinavir by allowing the virus to gain a foothold through the development of resistance. An understanding of these issues will be critical in realizing the full potential of this potent new drug for the control of HIV-1 infection. PMID- 9873803 TI - HIV treatment--a rational approach to the use of antiretroviral agents. AB - Over the last 2 years, advances in many areas of HIV clinical research and data on the effectiveness of potent combination therapy have substantially influenced the overall perspective of long-term management of HIV disease. However, the progress against HIV also came as a result of the better understanding of HIV pathogenesis. Research work in basic science has contributed considerably to obtain a clearer picture of the mechanisms of HIV infection, mainly through the understanding of key steps in the dynamics and kinetics of viral replication in vivo. Molecular biology also revealed much about the mechanisms of HIV virulence and the emergence of drug resistance. This article will give a short overview of the most recent advances in the field. PMID- 9873804 TI - Prion diseases. AB - Although the nature of the infectious agent causing prion diseases is still debated, several of its molecular characteristics have been clarified in remarkable detail. The transmissibility of bovine spongiform encephalopathy to humans dramatically highlights the need for research focused at interference with prion replication and spread, and at prevention of brain damage. Precondition to achieving these goals is a thorough understanding of prion biology, and in particular of its protein chemistry. PMID- 9873805 TI - Prions and haemophilia: assessment of risk. AB - Based on information accumulated to date, it is still difficult to assess the risk of Creutzfeldt-Jakob disease (CJD) and blood transfusion with any degree of confidence. However, it is reasonable to conclude that CJD is produced by a transmittable agent which is probably contained in low titer in the blood of infected people and animals. From the present clinical and epidemiological studies, transmission by blood or blood products appears to be a rare or non existent cause of current and past cases of CJD in humans. Since blood products are necessary to prevent the immediate risk of death or significant morbidity in many clinical conditions, therapeutic decisions should be made after consideration of the known risk in these situations vs the theoretical long-term risk of the rare occurrence of CJD. PMID- 9873806 TI - Discrepancies in potency assessment of recombinant FVIII concentrates. AB - Results of assays of recombinant FVIII concentrates have been reviewed over a 10 year period. Initially there was wide variability between laboratories but this was minimised by the development of standardised assay methodology, in particular the use of haemophilic plasma for pre-dilution and 1% albumin in assay buffers. Using this standardised methodology and concentrate standards, there were no major differences in potency between one-stage, two-stage and chromogenic assays on the two full-length recombinant FVIII concentrates. However, using a plasma standard, the chromogenic method gave much higher potencies than the one-stage method on the same concentrates, and this explains a similar discrepancy found in patients' post-infusion samples after injection of recombinant concentrates. It is suggested that concentrate standards be used for such post-infusion samples in order to minimise this discrepancy. PMID- 9873807 TI - In vivo recovery with products of very high purity--assay discrepancies. AB - In view of reports of FVIII assay discrepancies in post-infusion plasma samples depending on methods used, we compared FVIII results run by each of four different methods following infusion of rFVIII (Kogenate). Nine persons with haemophilia A were infused with each of two lots of product. Plasma samples were obtained at baseline, and at 10 min, 30 min, 1, 2, 4, 8, 12, 14, 30 and 48 h post infusion for measurement of FVIII. FVIII assay methods were chromogenic, and one stage APTT using three different types of activators: micronized, silica, ellagic acid, and kaolin. The same reference plasma standard was used throughout. Results demonstrated a consistent difference in FVIII values, with chromogenic assays being considerably higher than those run by one-stage assays. The discrepancy was greatest when kaolin was the activator. These results point out the problems in attempting to determine the "correct" FVIII level in patient plasma samples following infusion of high purity FVIII preparations. Potential "pitfalls" include the standard used for defining product potency, the methods, reagents, instrumentation and standards used in assaying plasma samples and, in some instances, the characteristics of the product itself. This situation has considerable cost implications, potential impact on patient care, and makes it difficult to compare results between laboratories. PMID- 9873808 TI - Influence of phospholipids on the assessment of factor VIII activity. AB - In view of reported discrepancies between different factor VIII assays, the influence of phospholipids on the performance of one-stage clotting (OS) and chromogenic substrate (CS) assays was evaluated. The B domain deleted recombinant factor VIII, rVIII SQ, two full-length recombinant products and a plasma derived factor VIII concentrate were each diluted into severe haemophilia A plasma and assayed against a plasma standard. The one-stage activity was 50, 80, 75 and 106%, respectively, of the chromogenic result. Variations in the phospholipid concentration did not affect the chromogenic assay, except at very low levels where the apparent activity increased. In contrast, dilution of the phospholipid reagent had a substantial influence on the activity measured by OS assays, especially in the case of rVIII SQ. At low levels of phospholipid, the one-stage activity of rVIII SQ exceeded the chromogenic result. When mixtures of phosphatidylserine (PS) and phosphatidyl-choline (PC) were used as a source of phospholipid, the OS results for rVIII SQ agreed well with the CS activity as long as the content of PS was below 10%, i.e., closer to the physiological level. At higher levels of PS, as in most commercial APTT reagents, the OS activity decreased. When the APTT reagent was replaced by platelets in the OS assay, the results compared well with those obtained by the CS assay for both t-VIII SQ and full-length factor VIII products. PMID- 9873809 TI - Quality control and factor VIII assays. AB - Quality assurance is a vital prerequisite of good laboratory practice. The application and continuous scrutiny of adequate internal quality control measures are essential to ensure that laboratory performance is consistent from day to day. External quality assessment is particularly important to verify the accuracy and reliability of laboratory results Consequently, without appropriate IQC and EQA, laboratory results can have no true validity. PMID- 9873810 TI - Diagnosis of von Willebrand disease. AB - von Willebrand disease (vWD) is a bleeding disorder caused by quantitative or qualitative defects of von Willebrand factor (vWF). vWF is synthesized by endothelial cells and megakaryocytes and circulates in plasma as a multimeric high molecular weight glycoprotein. vWF plays a major role in the early phases of ostasis by promoting platelet-vessel wall and platelet-platelet interactions under high shear conditions. It is also the carrier of coagulation factor VIII (FVIII) in plasma. A deficiency of vWF results in impairment of both primary and secondary phases of ostasis. Therefore, patients with vWD manifest bleeding symptoms that are typical of defects of primary ostasis (mucocutaneous haemorrhages) but, in case of severe deficiency of vWF, there are also haemarthroses and haematomas, which are typical of those seen with coagulation defects. Several types and subtypes of vWD have been described with a high degree of heterogeneity. The diagnosis is based on measurements of plasma and platelet vWF, the ability of vWF to interact with its platelet receptor and the analysis of the multimeric composition of vWF. Due to the heterogeneity of vWF defects, a correct diagnosis of types and subtypes may be sometimes difficult but is very important for an appropriate treatment of patients with vWD. PMID- 9873811 TI - Treatment of von Willebrand disease. AB - von Willebrand disease is the most frequent of inherited bleeding disorders (1:100 affected individuals in the general population). The aim of treatment is to correct the dual defects of haemostasis, i.e., abnormal coagulation expressed by low levels of factor VIII and abnormal platelet adhesion expressed by a prolonged bleeding time. There are two main options available for the management of von Willebrand disease: desmopressin and transfusion therapy with blood products. Desmopressin is the treatment of choice in patients with type 1 von Willebrand disease, who account for approximately 80% of cases. This pharmacological compound raises endogenous factor VIII and von Willebrand factors and thereby corrects the intrinsic coagulation defect and the prolonged bleeding time in most type 1 patients. In type 3 and in the majority of type 2 patients desmopressin is not effective, and it is necessary to resort to plasma concentrates containing factor VIII and von Willebrand factor. Treated with virucidal methods, these concentrates are effective and currently safe, but the bleeding time defect is not always corrected by them. Platelet concentrates or desmopressin can be used as adjunctive treatments when poor correction of the bleeding time after concentrates is associated with continued bleeding. PMID- 9873812 TI - Females with von Willebrand disease: 72 years as the silent majority. AB - The monthly challenge of menstruation as well as the haemostatic challenge of childbirth postpartum renders more females than males symptomatic with von Willebrand disease. Among vWD patients, the obstetrical and gynaecological morbidity is certainly more pronounced in Type 2,3 patients compared to Type 1 patients, but even in the latter group there is a high proportion of menorrhagia with associated anaemia, loss of time from work/school and the use of hysterectomy for ultimate control of bleeding. Despite the well known adage of the "gestational palliation" of vWD, there is a high proportion of postpartum haemorrhage in Type 1 patients also especially after the first 24 h after delivery. This may occur despite normalization of the factor VIIIc level in the third trimester, particularly in Type 2,3 patients. With the increasing availability of intranasal/subcutaneous DDAVP that could be readily administered at home for menorrhagia, there recently has been ongoing efforts internationally to determine the prevalence of vWD in females presenting with menorrhagia with a prevalence of 17% combined from two studies of 180 patients total. Issues remain regarding the optimal dose/schedule of intranasal/subcutaneous DDAVP for menorrhagia and the relative efficacy of antifibrinolytic agents. The proper role of oral contraceptives and danazol also deserves further study in vWD patients with menorrhagia. In sum, a comprehensive care approach in females with vWD is warranted analogous to the successful model of care of male haemophiliacs with the intent to (a) reduce unnecessary surgical interventions for menorrhagia, (b) improve the quality of life during menses and (c) optimize peri-partum management. PMID- 9873813 TI - Combined factors V and VIII deficiency--the solution. AB - Combined deficiency of coagulation factor V and factor VIII is an autosomal recessive disorder which has been observed in a number of populations around the world. However, this disease appears to be most common in the Mediterranean basin, particularly in Jews of Sephardic and Middle Eastern origin living in Israel. We have taken a positional cloning approach toward identifying the gene responsible for this disorder. We initially studied 14 affected individuals from nine unrelated Jewish families using a panel of polymorphic genetic markers spaced throughout the human genome. The combined factors V and VIII deficiency gene was mapped to a locus on the long arm of chromosome 18 with a maximal LOD score of 13.22. A detailed genetic analysis identified two distinct haplotypes among these families, suggesting two independent founders or, alternatively, a single ancient founder with a more recent split of these subpopulations. Further work to identify and characterize the gene responsible for combined factors V and VIII deficiency should provide important insights into the biosynthesis of these homologous proteins. PMID- 9873814 TI - The management of factor XI deficiency. AB - Factor XI deficiency leads to a more variable bleeding tendency than haemophilia A or B. Although severely deficient individuals are likely to bleed excessively especially after surgery in areas of the body with increased fibrinolysis, there is evidence that some partially deficient individuals are at risk of excessive bleeding. This will entail careful planning for surgery. Several therapeutic modalities are available which include fresh frozen plasma, factor XI concentrates, fibrin glue, antifibrinolytic drugs and desmopressin. The advantages and risks of these are considered. Factor XI concentrate may be indicated for procedures with a significant risk of bleeding especially in younger patients with severe deficiency, but its use in older patients has been associated with thrombotic phenomena. If fresh frozen plasma is to be used, it is preferable to obtain one of the virally inactivated products. Fibrin glue is a useful treatment which deserves further study. PMID- 9873815 TI - Clinical picture and treatment strategies in factor VII deficiency. AB - Factor VII is a trace protein required for normal haemostasis. Deficiency of factor VII comprises a highly heterogeneous disease group. Factor VII deficiency can cause bleeding, in particular if factor VII is extremely low, but a few cases lacking factor VII function entirely or subtotally may not present with a history of bleeding. Bleeding problems are not often reported in patients having a factor VII:C level at 10-15% of normal or more. Bleeding is frequently of mucocutaneous type, but the whole array of haemophilic bleeding may also occur. To control bleeding, during surgery in particular, substitution is required in the severe case of factor VII deficiency, but clinical studies documenting which correctional levels of factor VII:C to aim are lacking. It appears that a critical low level (trough) value at 10-15% may be anticipated, but clear documentation does not exist. Substitution programmes may include plasma or plasma derived factor IX concentrates of lower degrees of purity, so-called prothrombin complex concentrates that also are relatively impure, and pure factor VII concentrates. An alternative is a recombinant factor VIIa molecule. However, this concentrate has not received license in a number of countries. Thrombotic manifestations appear to occur more often than expected in the factor VII deficient patients, some have been linked to the use of impure concentrates, others to preexisting thrombophilic risk factors, but some are unexplained and may bear a relationship to the deficiency state of factor VII itself. Controlled clinical trials are highly warranted in this rare bleeding condition. PMID- 9873816 TI - Cytokine regulation of expression of class I MHC antigens. PMID- 9873817 TI - A mechanism of differential expression of GLUT2 in hepatocyte and pancreatic beta cell line. AB - DNase I footprinting assay using liver nuclear extracts revealed six protected regions between nucleotide -600 and +110 and hence named Box I-VI. Upstream promoter element (UPE), a DNA element playing crucial role in transcriptional control of the tissue specific expression of pancreatic beta-cell, has been detected within the proximal region of rat GLUT2 promoter. This region is included in Box VI. The protein-DNA interaction in this region (Box VI) was confirmed by mobility shift assay using liver nuclear extracts. Deletion of the region between -585 bp and -146 bp resulted in dramatic changes in promoter activity when they were expressed in liver and beta-cell derived cell line. When 585/-146 construct was expressed in liver, the activity was decreased to 46%, whereas the activity in beta-cell line, HIT-T15 cell, was increased by 84% when compared to -146/+190 construct. These opposing phenomena can be explained by the fact that beta-cell specifically expresses the UPE binding protein. Assuming that there may be Box VI-binding protein playing negative roles both in hepatocyte and beta-cell, and that the protein acts as a negative regulator of GLUT2 gene, the UPE binding protein in the beta-cell may overcome the inhibition by binding to the protein. PMID- 9873818 TI - Changes of phospholipase D activity in TNF-alpha and anti-Fas/Apo1 monoclonal antibody induced apoptosis in HL-60 and A20 cells. AB - The changes of phospholipase D (PLD) activity were investigated during the courses of apoptotic process induced by tumor necrosis factor (TNF)-alpha or anti Fas/Apo1 antibody in human premyelocyte HL-60 and murine B cell lymphoma A20 cells. The treatment of recombinant TNF-alpha to HL-60 cells resulted in the increased PLD activity as determined by the phosphatidylethanol formation in the presence of 1% ethanol. The enhancement of PLD activity was also observed in the anti-Fas/Apo1 monoclonal antibody-treated A20 cells. However, the activity of PLD was maximized when HL-60 and A20 cells were treated with either TNF-alpha or anti Fas/Apo1 monoclonal antibody for 6 h. Both TNF-alpha and anti-Fas/Apo1 monoclonal antibody increased PLD activity in a dose-dependent manner up to 200 U/ml and 200 ng/ml, respectively. When the intracellular activity of protein kinase C (PKC) was interrupted by treatment of calphostin-C, both the PLD activation and the apoptosis induced by TNF-alpha and anti-Fas/Apo1 monoclonal antibody appeared to be inhibited. Since PKC is reported to activate PLD, the results indicate that the intracellular signaling cascade via PLD may play a role in the induction of apoptosis induced by TNF-alpha and anti-Fas/Apo1 monoclonal antibody. PMID- 9873819 TI - Changes in the expression of c-myc, RB and tyrosine-phosphorylated proteins during proliferation of NIH 3T3 cells induced by hyaluronic acid. AB - We have shown that hyaluronic acid stimulates the proliferation of quiescent NIH 3T3 cells. We have shown that treatment of 1 mg/ml hyaluronic acid results in increase of tyrosine phosphorylation of two proteins, MW 124 kDa and 60 kDa as detected by anti-tyrosine antibodies by Western blot analysis. Maximum phosphorylation occurred within 2 h after addition of 1 mg/ml hyaluronic acid. Stimulation of proliferation was also accompanied by increase in c-Myc protein, which was inhibited by amlloride, an inhibitor of Na+/H+ antiporter and EGTA and increase in the steady state level of pRb, the RB gene product. These results suggest that the intracellular signal transduction pathways that mediate the stimulatory effects of hyaluronic acid on cellular proliferation are similar to those of growth factors. PMID- 9873820 TI - Telomerase is strongly activated in hepatocellular carcinoma but not in chronic hepatitis and cirrhosis. AB - Telomerase is highly activated in human immortal cell lines and tumor tissues, whereas it is not activated in primary cell strains and many tumor-adjacent tissues. It is suggested that telomerase activation is one of the critical steps in malignant transformation. In the present study, the telomerase activity was investigated in hepatocellular carcinoma tissues and non-tumor liver tissues from Korean patients with chronic hepatitis and cirrhosis. Eighty two liver tissues (24 chronic hepatitis specimens, 34 cirrhosis specimens, and 24 hepatocellular carcinomas) were obtained from 23 chronic viral hepatitis patients, 19 cirrhosis patients (including 7 liver transplants), and 24 patients with hepatocellular carcinoma, of which the surrounding non-tumor liver tissues were available in 16 patients (1 chronic hepatitis and 15 cirrhosis). As negative controls, 3 normal liver tissues were included. Protein from liver specimens was purified by a detergent lysis method as described elsewhere, and telomerase activity was measured in 2 diluents of each sample (1:1 and 1:100) by a telomeric repeat amplification protocol (TRAP). Telomerase was strongly activated in 79% (19/24) of the hepatocellular carcinomas, while weakly in 8% (2/24) of the chronic hepatitis tissues and in 24% (8/34) of the cirrhosis tissues. All of 3 normal control livers showed no telomerase activation. No relationship could be observed between the enhancement of telomerase activity and tumor nature. None of the chronic heaptitis or cirrhosis patients with mild telomerase activation in the liver have developed hepatocellular carcinoma for at least 2 years of follow-up period. These results suggest that the strong enhancement of telomerase activity may be a critical part of hepatocarcinogenesis, although the exact mechanism of such high activation in hepatocellular carcinoma is not clear. In addition, further study will be necessary to clarify the reason why no telomerase activity detectable by a conventional TRAP can be seen in some hepatocellular carcinoma. PMID- 9873821 TI - Production and characterization of monoclonal antibodies to oxidized LDL. AB - Oxidized low density lipoprotein (LDL) seems to take a part in atherogenesis through direct interactions with macrophages, endothelial cells, and smooth muscle cells, and is thought to participate in renal glomerular injury. For the purpose of illustrating the role of oxidized LDL in the human diseases, monoclonal antibodies were developed and characterized, recognizing oxidized LDL specific epitopes that do not exist on native LDL. LDL was oxidized by the incubation with CuSO4, and used as immunogen. Splenocytes from the immunized mouse and mouse myeloma cells were fused to produce hybridomas, which were screened for the secretion of oxidized LDL-specific antibodies. Immunoblot analysis and binding affinity assay showed that these monoclonal antibodies recognize malondialdehyde-conjugated peptide epitopes. PMID- 9873822 TI - Effect of petroleum ether extract of Panax ginseng roots on proliferation and cell cycle progression of human renal cell carcinoma cells. AB - Panax ginseng roots have long been used as a medicinal herb in oriental countries. We have investigated anti-proliferative effects of lipid soluble Panax ginseng components on human renal cancer cell lines. Petroleum ether extract of Panax ginseng roots (GX-PE) or its partially purified preparation (7:3 GX) was added to cultures of three human renal cell carcinoma (RCC) cell lines, A498, Caki-1, and CURC II. Proliferation of RCC cells was estimated by a [3H]thymidine incorporation assay and cell cycle distribution was analyzed by flow cytometry. GX-PE, 7:3 GX, panaxydol and panaxynol inhibited proliferation of all three RCC cell lines in a dose dependent manner in vitro with an order of potency, 7:3 GX > panaxydol > panaxynol = GX-PE. Additive effect of interleukin 4 was also demonstrated, most prominently in Caki-1 which responded poorly to GX-PE alone. Analysis of cell cycle in CURC II and Caki-1 treated with GX-PE demonstrated increase in G1 phase population and corresponding decrease in S phase population. The present study demonstrated that proliferation of human RCC cell lines were inhibited by lipid soluble components of Panax ginseng roots by blocking cell cycle progression at G1 to S phase transition. PMID- 9873823 TI - Purification and cloning of glyoxalase II from rat liver. AB - Glyoxalase (GLO) II, which is a component of GLO system and catalyze the conversion of S-lactoyl-glutathione to D-lactate, was purified 1488 fold from rat liver by two steps of Affigel blue and carbobenzoxyglutathione-Sepharose 4B affinity chromatography. The molecular weight of the enzyme was estimated to be 29 kDa which is similar to those from other species. The sequence of N-terminal 9 amino acid residues was determined to be MGIRLLPAT. This was then used to synthesize degenerative primers. cDNA clone was isolated by first synthesizing cDNA from RNA and then PCR amplification. The sequence of cDNA clone was determined by serial sequencing analysis. PMID- 9873824 TI - Interferon-gamma upregulates the stromelysin-1 gene expression by human skin fibroblasts in culture. AB - The equilibrium between deposition and degradation of extracellular matrix(ECM) is essential to normal tissue development and repair of wound or inflammatory responses. It has recently become apparent that several cytokines and growth factors are capable of modulating fibroblast proliferation and biosynthetic activity. To understand the role of these factors in connective tissue regulation, we examined the effect of interferon-gamma (IFN-gamma) on stromelysin 1 gene expression in cultured human dermal fibroblasts. The steady-state levels of stromelysin-1 mRNA were increased in IFN-gamma treated cultured dermal fibroblasts. In the CAT assay, the stromelysin-1 promoter activity was increased 2.8-fold compared with untreated control. Therefore IFN-gamma stimulates the stromelysin-1 promoter activity, resulting in transcriptional enhancement of gene expression. Transforming growth factor-beta (TGF-beta) showed the antagonistic action to the effects of IFN-gamma in cultured dermal fibroblasts. Furthermore, gel mobility shift assays demonstrated enhanced AP-1 binding activities in nuclear extracts from cells incubated with IFN-gamma. These data suggest that IFN gamma is an up-regulator and TGF-beta is a down regulator on the stromelysin-1 gene expression, respectively, and the AP-1 binding site may be necessary for gene response. PMID- 9873825 TI - Expression of a recombinant branched chain alpha-oxo acid dehydrogenase complex E2 (BCOADC-E2) in insect cells and its immunoreactivity to autoimmune sera. AB - Preparation of a pure autoantigen by way of recombinant DNA technology has an important value in an accurate diagnosis or prognosis of an autoimmune disease. BCOADC-E2 subunit, a mitochondrial protein, has been known to be the autoantigen of primary biliary cirrhosis (PBC), a chronic autoimmune liver disease, as well as idiopathic dilated cardiomypathy (IDCM), a chronic autoimmune heart disease. Recombinant form of this molecule had been expressed in E. coli but with low yield and severe degradation. Furthermore, sera from IDCM patients failed to recognized BCOADC-E2 molecule produced in prokaryotic expression system. In this study, a recombinant bovine BCOADC-E2 fusion protein has been expressed in insect cells using baculovirus expression system and analyzed anti-BCOADC-E2 reactivity in sera from patients with PBC or with IDCM. Optimal production of the recombinant fusion protein has been achieved at 20 multiplicity of infection (MOI), and the protein was affinity-purified using metal-binding resins. The affinity-purified BCOADC-E2 protein was successfully recognized by sera from PBC patients, but not by sera from IDCM patients suggesting that the different auto immune response against BCOADC-E2 is needed to be elucidated in terms of epitope recognition. PMID- 9873826 TI - Rapid increase of cytosolic content of acetyl-CoA carboxylase isoforms in H9c2 cells by short-term treatment with insulin and okadaic acid. AB - Mammalian acetyl-CoA carboxylase (ACC) is present in two isoforms, alpha and beta, both of which catalyze formation of malonyl-CoA by fixing CO2 into acetyl CoA. ACC-alpha is highly expressed in lipogenic tissues whereas ACC-beta is a predominant form in heart and skeletal muscle tissues. Even though the tissue specific expression pattern of two ACC isoforms suggests that each form may have a distinct function, existence of two isoforms catalyzing the identical reaction in a same cell has been a puzzling question. As a first step to answer this question and to identify the possible role of ACC isoforms in myogenic differentiation, we have investigated in the present study whether the expression and the subcellular distribution of ACC isoforms in H9c2 cardiac myocyte change so that malonyl-CoA produced by each form may modulate fatty acid oxidation. We have observed that the expression levels of both ACC forms were correlated to the extent of myogenic differentiation and that they were present not only in cytoplasm but also in other subcellular compartment. Among the various tested compounds, short-term treatment of H9c2 myotubes with insulin or okadaic acid rapidly increased the cytosolic content of both ACC isoforms up to 2 folds without affecting the total cellular ACC content. Taken together, these observations suggest that both ACC isoforms may play a pivotal role in muscle differentiation and that they may translocate between cytoplasm and other subcellular compartment to achieve its specific goal under the various physiological conditions. PMID- 9873827 TI - A new member of alpha 1-adrenoceptor-coupled G alpha h (transglutaminase II) family in pig heart: purification and characterization. AB - We previously reported an identification of a 77-kDa GTP-binding protein that co purified with the alpha 1-adrenoceptor following ternary complex formation. In the present paper, we report on the purification and characterization of this GTP binding protein (termed G alpha h5) isolated from pig heart membranes. After solubilization of pig heart membranes with NaCl, G alpha h5 was purified by sequential chromatographies using DEAE-Cellulose, Q-Sepharose, and GTP-agarose columns. The protein displayed high-affinity GTP gamma S binding which is Mg(2+) dependent and saturable. The relative order of affinity of nucleotide binding by G alpha h5 was GTP > GDP > ITP >> ATP > or = adenyl-5'-yl imidodiphosphate, which was similar to that observed for other heterotrimeric G-proteins involved in receptor signaling. Moreover, the G alpha h5 demonstrated transglutaminase (TGase) activity that was blocked either by EGTA or GTP gamma S. In support of these observations, the G alpha h5 was recognized by a specific antibody to G alpha h7 or TGase II, indicating a homology with G alpha h (TGase II) family. These results demonstrate that 77-kDa G alpha h5 from pig heart is an alpha 1 adrenoceptor-coupled G alpha h (TGase II) family which has species-specificity in molecular mass. PMID- 9873828 TI - Suppression of multidrug resistance via inhibition of heat shock factor by quercetin in MDR cells. AB - MDR1 promoter has been shown to contain heat shock elements (HSE), and it has been reported that FM3A/M and P388/M MDR cells show a constitutively activated heat shock factor (HSF), suggesting that HSF might be an important target for reversing the multidrug resistance. Therefore, it was examined whether quercetin, which has been shown to interfere with the formation of the complex between HSE and HSF, and to downregulate the level of HSF1, can sensitize MDR cells against anticancer drugs by inhibition of HSF DNA-binding activity. In this study, quercetin appeared to inhibit the constitutive HSF DNA-binding activity and the sodium arsenite-induced HSF DNA-binding activity in the MDR cells. The basal and sodium arsenite-induced MDRCAT activities were remarkably suppressed by the treatment of quercetin. These results were well consistent with the finding that the treatment of quercetin decreased the expression level of P-gp, MDR1 gene product, in dose-dependent manner, and markedly increased the sensitivity of MDR cells to vincristine or vinblastine. These results suggest that quercetin can decrease the expression of P-gp via inhibition of HSF DNA-binding activity, and might be useful as a chemosensitizer in MDR cells. PMID- 9873829 TI - Partial purification and characterization of a novel murine factor that augments the expression of class I MHC antigens on tumor cells. AB - A soluble factor which augments the expression of major histocompatibility complex class I (MHC I) antigens on a number of murine tumor cell lines, has been isolated from the culture supernatants of mixed lymphocyte reaction of spleen cells derived from C57B1/6, Balb/c and Swiss mice. The factor, termed MHC augmenting factor (MHC-AF) has been partially purified by Sephadex G-100 column chromatography and reverse phase HPLC. MHC-AF activity is associated with an 18 kDa molecule. MHC-AF activity was resistant to pH 2.0 treatment and partially purified MHC-AF preparations did not have any activity in L929 cell/vesicular stomatitis virus (VSV) interferon bioassay system. Antibodies to IFN-gamma did not block the activity of MHC-AF. These results indicate that a MHC-AF distinct from IFN-gamma, is produced by mouse spleen cells undergoing a mixed lymphocyte reaction. PMID- 9873831 TI - Alterations of CDKN2 (MTS1/p16INK4A) gene in paraffin-embedded tumor tissues of human stomach, lung, cervix and liver cancers. AB - The CDKN2 (MTS1/p16INK4A) gene, encoding cyclin dependent kinase inhibitor, was found to be homozygously deleted at a high frequency in cell lines from many different types of cancer and some primary cancers. To determine the frequency of CDKN2 mutations in most common human cancers in Korea, PCR and PCR-SSCP analyses for the exon 2 of CDKN2 were performed on each set of 20 formalin-fixed and paraffin-embedded tumor tissues of stomach adenocarcinomas, lung cancers, cervix cancers and hepatocellular carcinomas. No mutations in exon 2 of CDKN2 were found in 20 stomach adenocarcinomas. In contrast to rare mutations in stomach adenocarcinomas, a high frequency of CDKN2 mutations was identified in other 3 cancers, 11 of 20 (55%) lung cancers (7 of 10 NSCLCs and 4 of 10 SCLCs), 14 of 20 (70%) cervix cancers and 11 of 20 (55%) hepatocellular carcinomas. These results suggest that mutations of the CDKN2 gene might be an important genetic change in NSCLCs, cervix cancers and hepatocellular carcinomas. PMID- 9873830 TI - Molecular biological characterization of enterovirus variant isolated from patients with aseptic meningitis. AB - In Korea, there was a big outbreak of aseptic meningitis in 1993. Six clinical isolates of enterovirus were obtained from patients with aseptic meningitis and were identified as echovirus type 9 by serotyping with a pool of neutralizing antisera. For molecular characterization of the isolates, the nucleotide sequences of 5'-noncoding region (NCR), VP4, VP2, VP1, 2A and 2C regions of the isolates were compared with the corresponding regions of echovirus type 9 Hill and Barty strains. Unlike Hill strain, Barty strain contained a C-terminal extension to the capsid protein VP1 with an RGD (argnine-glycine-aspartic acid) motif. To determine whether similar structural features were present in our isolates, their nucleotide sequences including the VP1 region were analyzed. All isolates exhibited the VP1 extension with the RGD motif. We concluded the Korean isolates in the year of 1993 as the echovirus type 9 Barty strain although the isolates showed 15-20% nucleotide sequence differences in the several genomic regions. PMID- 9873833 TI - Molecular biology of neuronal voltage-gated calcium channels. PMID- 9873832 TI - Heterogeneous HBV mutants coexist in Korean hepatitis B patients. AB - Although many hepatitis B virus (HBV) mutants have been found in all open reading frames since the precore defective mutant was initially reported, systematic investigations of diverse HBV mutant populations in hepatitis B patients have not been performed. Therefore, we examined whether heterogeneous mutant populations simultaneously exist in Korean hepatitis B patients. In order to detect hepatitis B virus mutants, we amplified a conserved core region and a surface antigen region of HBV DNA by PCR from sera of 27 Korean chronic hepatitis B patients, and then performed single strand conformational polymorphism analysis followed by DNA sequencing analysis. The results showed that heterogeneous HBV mutants in both regions were present in a single as well as in various hepatitis B patients. Sequence analysis revealed a defective interfering particle with missense mutation in the core region. We also found that two subtypes of adr and adw coexisted in a single patient. In addition, a point mutation causing a stop codon in the surface antigen region was observed. We are further analyzing the clinical implications of HBV mutants to identify their roles in the pathogenesis of chronic hepatic disorders induced by HBV. PMID- 9873834 TI - A variant of ornithine aminotransferase from mouse small intestine. AB - The ornithine aminotransferase (OAT) activity of mouse was found to be highest in the small intestine. The mitochondrial OAT from mouse small intestine was purified to homogeneity by the procedures including heart treatment, ammonium sulfate fractionation, octyl-Sepharose chromatography, and Sephadex G-150 gel filtration. Comparing to the amino acid sequence of mouse hepatic OAT, six N terminal amino acid residues have been deleted in intestinal OAT. However, the subsequent sequence was identical with that of hepatic OAT. The molecular weights of both intestinal and hepatic OAT were estimated as 46 kDa by SDS-gel electrophoresis and as 92 kDa by gel filtration, indicating that both native OATs are dimeric. Biochemical properties of intestinal OAT, such as molecular weight, pH optimum and K(m) values for L-ornithine and alpha-ketoglutarate, were similar to those of hepatic OAT. However, intestinal OAT was more labile than hepatic OAT to tryptic digestion. PMID- 9873835 TI - Role of Ca2+ influx in the tert-butyl hydroperoxide-induced apoptosis of HepG2 human hepatoblastoma cells. AB - Oxidative stress appears to be implicated in the pathogenesis of various diseases including alcoholic liver injury. In this study we investigated the mechanism of apoptosis induced by tert-butyl hydroperoxide (TBHP) in HepG2 human hepatoblastoma cells. Treatment with TBHP significantly reduced glutathione content and glutathione reductase activity, and increased glutathione peroxidase activity, indicating that TBHP induced oxidative stress in the HepG2 cells. TBHP also induced reduction of cell viability and DNA fragmentation, a hallmark of apoptosis, in a dose-dependent manner. In addition, TBHP induced a sustained increase in intracellular Ca2+ concentration, which was completely prevented by the extracellular Ca2+ chelation with EGTA. TBHP also induced Mn2+ influx. These results indicate that the intracellular Ca2+ increase by TBHP is exclusively due to Ca2+ influx from the extracellular site. Treatment with either an extracellular (EGTA) or an intracellular Ca2+ chelator (BAPTA/AM) significantly suppressed the TBHP-induced apoptosis. Taken together, these results suggest that TBHP induced the apoptotic cell death in the HepG2 cells and that Ca2+ influx may play an important role in the apoptosis induced by TBHP. PMID- 9873837 TI - Caffeine causes glycerophosphorylcholine accumulation through ryanodine inhibitable increase of cellular calcium and activation of phospholipase A2 in cultured MDCK cells. AB - Glycerophosphrylocholine (GPC) is a renal medullary compatible organic osmolyte that is derived from choline via phosphatidylcholine, which is catalyzed in part by phospholipase A2 (PLA2) and its degradation by GPC: choline phosphodiesterase (GPC: choline PDE). We found that caffeine elevated intracellular free calcium ([Ca2+]i) and GPC level in cultured MDCK cells, canine kidney epithelial cells, and propose a possible biochemical mechanism. When MDCK cells were incubated for 3 h with 1 to 10 mM caffeine, cellular GPC was elevated in a dose-dependent manner, and this occurred independently of the extracellular osmolality. Caffeine stimulated the rate of [14C]choline incorporation into [14C]GPC and PLA2 activity. Whereas, GPC: choline PDE activity was accompanied by less of increase. These enzyme changes demonstrate the increased net synthesis of MDCK GPC. In order to identify what triggers the PLA2 activation, [Ca2+]i was measured by using a fluorescence dye, Fura-2. Caffeine (10 mM) resulted in a typical transient increase in MDCK [Ca2+]i concentration, and this increase was greatly inhibited by pretreatment of MDCK cells with 10 mM ryanodine for 5 min. Ryanodine (10 mM) also inhibited the caffeine-induced stimulation of PLA2 activity. These findings provide the first evidence that caffeine in MDCK cells causes a ryanodine-inhibitable increase of [Ca2+]i and PLA2 activity, resulting in cellular GPC accumulation. PMID- 9873838 TI - Endoplasmic reticulum retention and degradation of T cell antigen receptor beta chain. AB - The T cell antigen receptor-CD3 (TCR/CD3) complex is assembled in the endoplasmic reticulum (ER) of T cells after synthesis of individual chains, and is transported to the cell surface for immune recognition and regulation. Partially assembled or unassembled TCR chains are retained and rapidly degraded in the ER. These processes are strictly regulated in the ER at post-translational level for the maintenance of cellular homeostasis. In order to identify the region responsible for the ER retention and rapid degradation of the TCR beta chain, number of mutants were engineered and their fates, after synthesis in the ER of the HeLa cells, were investigated. Extensive mutagenic analysis of TCR beta chain, including changing the charged amino acid residues and two tyrosine residues of the transmembrane region into hydrophobic amino acid residues, did not alter the ER retention and rapid degradation. Soluble TCR beta chain and cytoplasmic tail truncation mutant were also rapidly degraded in the ER. However, N-glycosylation rate of soluble TCR beta chain in the ER was significantly increased possibly due to the increased exposure of the N-glycosylation site. These results suggest that the ER retention of TCR beta chain is mediated through its extracellular and transmembrane-cytoplasmic regions and that the rapid ER degradation can be caused by an exposure of unassembled subregion of TCR beta chain, either extracellular domain or hydrophobic transmembrane region to the hydrophilic environment (lumen of the ER) rather than by presence of a specific degradation signal. PMID- 9873836 TI - Cloning of adriamycin-resistant related (arr) gene in an adriamycin-resistant L1210 variant. AB - A partial fragment of novel sequence (arr, adriamycin-resistant related) was previously identified using the differential display (DD)-PCR technique with adriamycin-resistant L1210 variant (L1210AdR), which shows a typical multidrug resistant (MDR) phenotypes. The present research shows the isolation of full length arr cDNA sequence. To clone the full length cDNA of arr gene, DD-PCR fragments were subjected to 5'- and 3'-Rapid Amplification of cDNA End (RACE) method. The cloned arr cDNA consisted of 770 bases and contained an open reading frame of 153 bases, encoding a protein of 51 amino acid with the molecular mass of 4 kDa by in vitro translation reactions. Northern blot analysis showed that a 770 bases transcript arr gene was overexpressed in adriamycin-resistant L1210 variant, but not in the parent suggesting that the arr gene may be involved in the adriamycin-resistant phenotypes. PMID- 9873839 TI - Urinary N-acetyl-beta-D-glucosaminidase in rheumatoid arthritis. AB - Excretion of urinary N-acetyl beta-D-glucosaminidase (NAG) and its isoenzyme patterns were studied in two groups of patients with rheumatoid arthritis (RA) and in normal control subjects. Urine samples were collected from 30 seropositive RA patients, 19 seronegative RA patients, and 15 normal healthy subjects. All the patients and normal subjects were assessed to have normal liver and kidney functions. A small portion of the urine sample was dialyzed against 0.01 M phosphate buffer, pH 7.0 and NAG activity was monitored. Mean +/- SD values of urinary NAG in seropositive RA patients, in seronegative RA patients and in normal healthy subjects were found to be 4.20 +/- 3.73 U/g creatinine, 2.96 +/- 2.11 U/gm creatinine, and 1.71 +/- 0.6 U/g creatinine, respectively. The mean urinary, NAG value in RA patients was found to be significantly higher (P < 0.05) in seropositive RA compared to the mean NAG value in normal healthy subjects and patients with seronegative RA when analyzed by one way ANOVA and Tukey-HSD test. The mean proportion of isoenzyme form B to isoenzyme form A in seropositive RA patients was also found to be significantly different (P < 0.05) from the mean proportion of these forms in normal healthy subjects and seronegative RA patients. There also appears to be a correlation between the concentration of urinary NAG and severity of the disease in seropositive RA. PMID- 9873840 TI - Nickel (II)-induced apoptosis and G2/M enrichment. AB - Treatment with certain DNA-damaging agents induce a complex cellular response comprising pertubation of cell cycle progression and/or apoptosis on proliferating mammalian cells. Our studies were focused on the cellular effects of nickel (II) acetate, DNA-damaging agent, on Chinese hamster ovary (CHO) cells. Fragmented DNAs were examined by agarose gel electrophoresis and cell cycle was determined by DNA flow cytometry using propidium iodide fluorescence. Apparent DNA laddering was observed in cells treated with 240 microM nickel (II) and increased with a concentration-dependent manner. Treatment of nickel (II) acetate resulted in apoptosis which was accompanied by G2/M cell accumulation. Proportion of CHO cells in G2/M phase was also significantly increased in cells exposed to at least 480 microM nickel (II) from 57.7% of cells in the G0/G1 phase, 34.7% in the S phase, and 7.6% in the G2/M1 phase for 0 microM nickel (II), to 58.6%, 14.5%, and 26.9% for 640 microM nickel (II). These findings suggest that nickel (II) can modulate cellular response through some common effectors involving in both apoptotic and cell cycle regulatory pathways. PMID- 9873841 TI - Intraoperative transmission of blood-borne disease in haemophilia. AB - Because HIV and hepatitis B and C infection can be transmitted by exposure to infected blood or blood components, orthopaedists are at risk for HIV and hepatitis infection during surgical procedures. Thus, care must be taken to protect themselves from transmission of HIV and the hepatitis virus. The following precautions are recommended: double latex gloves, changed hourly, or a combination of cloth and latex gloves, enclosed hood and face-masks and operative isolator with umbilical-cord aspirator; knee-length impermeable gowns of high count polyester weave or plastic-lined nonwoven spun-lace polyester; a combination of shoe-covers that provide waterproof coverage as high as the knee; and disposable drapes. If a member of the operating team is inadvertently pricked or cut, the wound should be washed immediately with iodine, soap and water. If the injured person has been immunized for the hepatitis B (and has adequate titres), or is positive for hepatitis B surface antigen or antibody, no further treatment is necessary. Otherwise, two doses of hepatitis B immune globulin should be given, 5 mL immediately and 5 mL after 1 month. Because HIV infection can be transmitted by exposure to infected blood, orthopaedic surgeons are also at risk for HIV infection. Prospective studies suggest that this risk is very low; nevertheless, healthcare workers need to adhere rigorously to the aforementioned infection-control precautions to minimize the risk of exposure to blood. PMID- 9873842 TI - The Malmo-Klaipeda WFH twinning programme: a comparative description of the haemophilia cohorts. AB - In 1994, the Malmo-Klaipeda twinning programme was approved by the World Federation of Hemophilia. One of the first steps in the collaboration has been to set up a registry of the haemophilia patients in the Klaipeda area. In order to collect important clinical data the patients have been examined jointly by experts on haemophilia from the two centres. Seventeen out of 25 patients with severe haemophilia known at the Klaipeda centre were examined and compared to a matched cohort of patients from the Malmo centre. The main differences between the cohorts were that home treatment was not available to the Klaipeda patients, they received less treatment in general, had higher joint scores and more frequent bleeds. The pattern of transmission of blood-borne virus was very similar, with a high prevalence of hepatitis C antibodies. We conclude that the twinning programme between Malmo and Klaipeda has resulted in several achievements, including training of staff and a necessary inventory of the patients. This should not only form a suitable platform for the future development of haemophilia care in Lithuania, but could also serve as an example for liaisons between other haemophilia centres. PMID- 9873843 TI - Multidose pharmacokinetics of factor IX: implications for dosing in prophylaxis. AB - The aim of this study was to investigate the use of single-dose pharmacokinetic data for factor IX (FIX) to predict multidose pharmacokinetics and explore their use for pharmacokinetic dosing in prophylactic treatment of haemophilia B. Eight patients with severe haemophilia B were enrolled. Using single-dose pharmacokinetic data for each patient, plasma factor IX procoagulant activity (FIX:C) curves during prophylactic dosing were computer-simulated. The simulations were verified by repeated blood sampling and measurements of FIX:C. Theoretical dosing regimens to maintain a plasma trough level of 1.0 U dL-1 of FIX:C were calculated. A 2 x 2 week cross-over study on standard dosing according to bodyweight vs. dosing every three days based on individual pharmacokinetics was carried out. FIX:C was measured during each treatment period. FIX:C data from the plasma sampling generally confirmed the single-dose pharmacokinetic data used. Pharmacokinetically tailored dosing of FIX could result in considerable savings of factor concentrate as compared to current standard dosing. The study demonstrates the applicability of individual pharmacokinetics as a tool for cost effective utilization of FIX concentrates in the prophylactic treatment of haemophilia B. PMID- 9873844 TI - Long-term follow up of patients treated with intermediate FVIII concentrate BPL 8Y. AB - Long-term surveillance studies of clotting factor concentrates are important to detect infrequent or delayed complications and to provide data against which newer products can be compared. We have assessed the long-term use of BPL 8Y factor VIII (FVIII) concentrate (Bio Products Limited, Elstree, UK) in a cohort of 33 patients. These patients have been treated for a median of 96 months. They have received between one batch (in total) and 10 batches per year and between 1020 units (in total) and 116,700 units per year of BPL 8Y concentrate. No patient has developed a clinically significant FVIII inhibitor. There has been no evidence of transmission of hepatitis C, hepatitis B or HIV 1 or 2. Parvovirus B19 IgG antibody was present in 100% of the patients screened. Analysis of CD4 and CD8 lymphocyte subsets, using age-related normal ranges, showed persistently depressed values in five patients, one of whom had a consistently low CD4/CD8 ratio. PMID- 9873845 TI - Surgery in haemophilia: experience from a centre in India. AB - The present paper describes various kinds of surgery carried out with great success in 16 cases which included both severe and moderate haemophilia patients with modest amounts of factor concentrates and anti-fibrinolytic drugs. This is very important in developing countries where factor concentrates are not easily available. In one patient haemophilia was diagnosed only after surgery. None of the patients had inhibitor pre- or post-operatively. One patient who was HIV positive underwent orchidectomy successfully with only 6000 IU of factor VIII concentrate. PMID- 9873846 TI - Assay of factor VIII antigen (VIII:CAg) in 294 haemophilia A patients by a new commercial ELISA using monoclonal antibodies. AB - Monoclonal antibodies (MoAbs 833 and D4H1) directed against human factor VIII (FVIII) have been produced on a large scale to measure VIII:CAg by two-site ELISA (Asserachrom VIII:CAg, Diagnostica Stago). F(ab')2 from MoAb 833 were used for coating and bound VIII:CAg was revealed with MoAb D4H1 coupled to peroxidase. Control plasma (100 VIII:CAg U dL-1 by comparing with the International Standard) was used as reference. The assay sensitivity was 0.1 U dL-1 VIII:CAg. No apparent effect of the plasma proteins was observed provided plasma dilution was > or = 5. Thus this ELISA allowed us to estimate VIII:CAg levels of 0.5 U dL-1 in plasma. Levels of VIII:CAg were similar to those of VIII:C (correlation coefficient r = 0.87) in plasma from normal individuals (32 cases) and in patients with von Willebrand disease of various types (30 cases). Among 294 patients with haemophilia A (HA), 161 had severe HA (VIII:C < 1 U dL-1). Among those patients, 124 were cross-reacting material (CRM) negative with undetectable VIII:CAg and 37 were CRM+ (VIII:CAg 1-31 U dL-1). In 42 patients with moderate HA (VIII:C 1-5 U dL-1), 33 were CRM reduced (VIII:CAg 0.5-8 U dL-1) and nine were CRM+ with a VIII:CAg/VIII:C ratio of 6-91 (mean 34.3). In mild HA (91 cases with VIII:C > or = 6 U dL-1), 29 patients were classified as CRM+ (VIII:C 6-57 U dL-1, VIII:CAg 17 130 U dL-1 and VIII:CAg/VIII:C ratio 1.8-13.7 (mean 4.51)). In 62 CRM reduced patients there was a linear correlation between VIII:C (6-39 U dL-1) and VIII:CAg (2-36 U dL-1) levels (r = 0.88). In conclusion, this sensitive assay allows us to distinguish the quantitative CRM reduced and negative from the qualitative (CRM+) abnormalities in haemophilia A. PMID- 9873847 TI - Inhibitor development and substitution therapy in a developing country: Turkey. AB - Prevalence of inhibitor in developing countries, such as Turkey, where fresh frozen plasma (FFP) is still in use due to high cost of concentrates, is unknown. To determine the frequency of inhibitors in Turkish haemophiliacs exposed to blood products, 53 haemophilia A patients (age range 1-20; median: 11 years) and 12 haemophilia B patients (age range 3-20; median: 10 years), were evaluated; 31 haemophilia A patients (23 severe) received plasma-derived concentrates and 22 patients (10 severe) only FFP. No haemophilia B patients developed inhibitor, compared with seven of 53 (13%) haemophilia A patients, all with a severe defect (7/33; 21%) and treated with concentrates (7/23; 30%), whereas severe patients treated with FFP showed a lower risk to develop inhibitors (0/10, P = 0.07). Inhibitors were detected after 8-125 exposure days (median: 52). Intermediate purity concentrates and pasteurization seemed to be linked with a higher risk of inhibitor compared to high-purity concentrates and solvent-detergent inactivation for seven patients with inhibitor. In four of seven inhibitor patients low-dose concentrate was administered at 25 IU kg-1 twice weekly and inhibitor disappeared in 1-4 months. This regimen might be recommended for immune tolerance in developing countries for its lower cost. PMID- 9873848 TI - Increased bleeding in HIV-positive haemophiliacs treated with antiretroviral protease inhibitors. AB - Seventeen HIV-positive patients with congenital haemophilia, one with von Willebrand's disease, and one with acquired haemophilia in remission, were treated with antiretroviral protease inhibitors. Clear increases in the frequency of bleeds or changes in the bleeding pattern were documented in 10 individuals taking HIV protease inhibitors. Blood product requirements were increased in eight individuals with severe haemophilia over the 6 months after starting HIV protease inhibitors. Therapy with the HIV protease inhibitor was discontinued in two patients in whom the increased bleeding was severe. No haemostatic cause for the increased bleeding tendency could be identified. Patients with bleeding disorders should be warned about this potential complication of HIV protease inhibitor therapy, and should be closely monitored. PMID- 9873849 TI - Human immunodeficiency virus infection in haemophiliacs in Taiwan: the importance of CD4 lymphocyte count in the progression to acquired immunodeficiency syndrome. AB - From July 1984 to December 1996, we tested and studied 303 haemophilic patients for the infection of the human immunodeficiency virus (HIV). Among the 261 haemophilia A patients 44 were HIV positive (16.9%), while none of the haemophilia B patients was HIV positive. The mean age of the 44 HIV-seropositive patients in 1984 was 20.6 years (2-37 years). Seven who had known seroconversion dates and 29 whose first seropositive dates were known seroconverted before 1986. Acquired immunodeficiency syndrome (AIDS) has developed in 16 patients, nine of whom presented with Pneumocystis carinii pneumonia, three with tuberculosis infection, and 13 had died. The Kaplan-Meier estimate of the progression rate to AIDS after the date of first seropositive test is about 30% at the 10th year. The median survival time after development of AIDS obtained from the Kaplan-Meier estimate of the survival curve was 11.7 months. Statistical analysis for the covariate effects on the risk of developing AIDS by the Cox proportional hazards model revealed that there was a statistically significant negative association of the risk for progression to AIDS with the logarithm of initial CD4 cell counts (P = 0.027) and the rate of decline of CD4 cell counts (P = 0.040), but not with age (P = 0.650). In conclusion, the clinical characteristics of AIDS haemophiliacs in Taiwan were not different from that observed in western countries. Low initial CD4 cell count and sharp decline in CD4 cell counts, but not age, increased the risk of progression to AIDS. PMID- 9873850 TI - Haemophilia A: two cases showing unusual features at birth. AB - We report two patients with severe haemophilia A, factor VIII < 2 u dL-1, diagnosed at birth, through isolated bleeding in the adrenal gland (case 1) and through a haematoma in the liver (case 2). In these two clinical cases, the vital emergency, with haemorrhagic shock, required early diagnosis of the hereditary coagulation defect. Generally at birth, any unexplained bleeding should prompt screening for haemophilia. In the newborn period, the PTT is inadequate for many reasons and the laboratory evaluation must include factor VIII and IX levels. PMID- 9873851 TI - The surgical treatment of a haemophilic pseudotumour in an extremity: a report of three cases with pathological fractures. AB - A haemophilic pseudotumour is a rare complication of haemophilia occurring in 1 2% of patients with a factor VIII or IX deficiency. This report presents three surgical cases of pseudotumours involved in a pathological fracture in the extremities. All cases showed a favourable post-operative course. If the preoperative management is appropriately designed, a limb salvage operation for a pathological fracture due to a pseudotumour could be carried out successfully. Before choosing amputation of a limb, the surgeon should consider the possibility of limb salvage. PMID- 9873852 TI - Excision of a haemophilic pseudotumour of the ilium, complicated by fistulation. AB - We report the case of a patient with a haemophilic pseudotumour of the ilium who developed chronic fistulation, 6 months after en-bloc resection. During the initial resection, the large defect in the iliac wing was filled with bone cement, which may have contributed to fistula formation. A second surgical procedure consisted of excision of the fistula and bone cement and the dead space was obliterated by bringing the gluteus medius muscle into the defect. The fistula recurred, however. Reexcision of the fistula and obliteration of the dead space by a pedicled rectus abdominis muscle flap resulted in eradication of the fistula. We emphasize the importance of obliteration of dead space, resulting from large pseudotumour resection. The use of bone cement is not advocated. If a fistula does occur, a pedicled rectus abdominis muscle flap may be considered. PMID- 9873853 TI - Call for post-licensing human pharmacokinetic studies of administration of recombinant factor IX. PMID- 9873854 TI - Cryoprecipitate and the plastic blood-bag system. PMID- 9873856 TI - A meeting held in London, 12-13 January 1998, to discuss bleeding disorders in women. PMID- 9873855 TI - Cryoprecipitate and the plastic blood-bag system: provision of adequate replacement therapy for routine treatment of haemophilia. 1967. PMID- 9873857 TI - Circumcision and haemophilia: a perspective. PMID- 9873858 TI - The changing patterns of factor VIII (FVIII) and factor IX (FIX) clotting factor usage in a comprehensive care centre between 1980 and 1994. AB - The annual amount of clotting factor used by patients at the Royal Free Haemophilia Centre increased significantly from 4 million iu in 1980 to over 15 million iu by 1994 (P < 0.0001). In order to assess the reasons for this increase, data on concentrate usage over this period were retrospectively collected for patients who had haemophilia or von Willebrand's disease. Only patients who were registered exclusively at the Centre were included in the study. In total, 498 patients met the inclusion criterion. The median age of the cohort on 1 January 1980 was 21 (range < 1-69) years. During the period there were 88 births and 45 deaths. The majority of patients had haemophilia A (55%). The median follow-up period per patient was 2.1 (range 0-14.8) years. Despite adjusting for increases in the number of patients and changes in body weight, statistically significant increases in clotting factor usage were detected for some subgroups of patients, in particularly for those with severe haemophilia A and B and from the late 1980s onwards, for patients with von Willebrand's disease. Two reasons for this increase in clotting factor usage were identified as being the introduction of improved products and prophylaxis. However, the increased cost of clotting factor provision that has resulted from these changes in treatment policy should not be analysed in isolation but should be balanced off against cost decreases in other areas and against increases in the effectiveness of treatment. PMID- 9873859 TI - Contrast venography in young haemophiliacs with implantable central venous access devices. AB - To assess the risk of deep vein thrombosis in haemophiliacs with long-term central venous catheters, we studied haemophiliacs followed at our centre with implantable venous access devices (ports) in place for > 6 months. Medical records were reviewed for a history of catheter-related complications. Each patient was examined for physical stigmata of thrombosis. Patency of the vessels was evaluated by contrast venography. Of 21 males with ports, 19 had factor VIII deficiency and two factor IX deficiency. Nineteen ports were evaluable (i.e. were in place for > 6 months). Seventeen patients have their original ports in place; two ports were replaced for mechanical dysfunction (1) and recurrent infection (1). Difficulty withdrawing or infusing occurred with three ports, two of which were cleared with urokinase. Physical examination was normal on all 19 patients. Venograms were performed in 13 of 19 patients. Parents of the remaining six patients refused venography because of the need for peripheral venipuncture. One patient had a small nonocclusive thrombus on the same side as his functioning catheter, and another had minimal narrowing of the subclavian vein at the site of a prior catheter. The overall prevalence of clinically relevant upper venous system thrombosis identifiable by contrast venography was zero (95% CI, 0-23%). We conclude that haemophiliacs do not have as high a risk of thrombosis as other populations of patients with central venous catheters. The theoretical risk of thrombosis should not preclude use of central venous catheters in patients with haemophilia. PMID- 9873860 TI - Immunoadsorption for removal of inhibitors: update on treatments in Malmo-Lund between 1980 and 1995. AB - Treatment of severe bleeding and the performance of surgery in haemophilia patients with inhibitors creates severe problems. It is generally agreed that treatment is most effective if circulating levels of factor VIII/IX can be achieved long enough for control of haemostasis. Immunoadsorption with protein A for the removal of inhibitor has improved treatment for patients with initial inhibitor titres too high to neutralize by infusion alone. This is a summary of our experience in Malmo regarding immunoadsorption and haemostasis. A total of 19 applications with immunoadsorption in 10 patients were performed. On all occasions it was possible to eliminate totally the inhibitor or reduce it to low levels that could easily be neutralized with factor concentrate. Haemostatic levels of coagulation factors could be maintained for 5-9 days in all but one patient. This period was sufficient to stop ongoing haemorrhage or prevent excessive bleeding at surgical interventions. PMID- 9873861 TI - Zero incidence of inhibitor development in previously treated haemophilia A, HIV negative patients upon exposure to a plasma-derived high-purity and double viral inactivated factor VIII concentrate. AB - Thirty-six haemophilia A, HIV-negative, previously treated patients were changed therapy to a highpurity and double-inactivated (solvent/detergent and dry heating) previously unused factor VIII concentrate. The mean age of these patients was 27 years at the time of the change. Twenty-three patients were severe haemophiliacs (FVIII:C < 0.02 IU mL-1), seven moderate (FVIII:C between 0.02 and 0.05 IU mL-1) and six mild (FVIII:C > 0.05 IU mL-1). The mean follow-up with this single product was 16 months, with 82 accumulated exposure days and the mean consumption was 117,300 IU of FVIII corresponding to a mean of six batches per patient. No patient developed FVIII inhibitors (upper limit of the CI95: 7.98%), resulting in an incidence rate of 0/48 patient-years (upper limit of the CI95: 77/1000 patient-years). The change in therapy to this new factor VIII concentrate was not associated with the appearance of inhibitors. PMID- 9873862 TI - Alpha interferon therapy in Danish haemophiliac patients with chronic hepatitis C: results of a randomized controlled open label study comparing two different maintenance regimens following standard interferon-alpha-2b treatment. AB - Following a survey among all Danish haemophiliac patients 49 HIV-negative patients with chronic hepatitis C were offered enrollment in a randomized controlled open label study comparing two different maintenance regimens following standard interferon-alpha-2b treatment. Dose modifications and treatment discontinuation were based upon changes in transaminase levels. Forty seven patients enrolled received 3 MU of alpha interferon thrice weekly (TIW) for 3 months. Twenty-six nonresponders had their dose increased to 6 MU TIW for an additional 3 months, while 21 responding patients continued on 3 MU TIW. At 6 months, 25 patients with a complete or a partial biochemical response were randomly allocated to either a fixed dose regimen (13 patients) (3 or 6 MU thrice weekly) or an individualized dose regimen (12 patients) tapering interferon dose from 3 or 6 MU by one-third every 2 months if transaminases were persistently normal. The remaining 22 biochemical nonresponders were followed for an additional 6 months without further treatment. After 12 months of treatment, 18 patients (38%) had a virological response, irrespective of regimen, and seven patients (16%) had a sustained virological and biochemical response after 6 months of follow up. Overall, the individualized treatment regimen did not seem to offer any advantage over the fixed dose regimen. The response to alpha interferon treatment in Danish haemophiliac patients with chronic hepatitis C immediately after treatment is comparable to that obtained in previous studies among nonhaemophiliacs. However, a sustained virological and biochemical response was seen in only 16% of treatment patients. PMID- 9873863 TI - A longitudinal study of immunological status in Chinese haemophiliacs: importance of the heat viral inactivation of factor concentrates. II. Improvements of CD4/CD8 ratio after treatments with heat-inactivated factor concentrates. AB - Screened and heated clotting factor concentrates of intermediate purity have been used in Taiwanese haemophiliacs since the end of 1986. A significant improvement of CD4/CD8 ratio during the years 1987-1989 as compared with those during the years 1984-1986 was observed in haemophilia A patients [mean +/- SD (median), 1.191 +/- 0.495 (1.163) vs. 0.880 +/- 0.325 (0.838), P = 0.0020] who were seronegative for human immunodeficiency virus. Almost all patients received an increased amount of factor VIII concentrates and total plasma products since 1987. Multiple linear regression analysis for the association of CD4/CD8 ratio with changes in dosage of plasma products revealed that there was a significant positive association of CD4/CD8 ratio measured during 1987-1989 with dosage of factor VIII concentrate administered during 1984-1986 (P = 0.0230), which is an indicator for changes in viral load, but not with changes in dosage of plasma products, which are indicators for changes in plasma protein intake. Our data indicate that immunological abnormalities after replacement therapy observed in haemophiliacs are mainly attributed to virus infection through infusion of factor concentrates, not to allogeneic proteins existing in plasma products. PMID- 9873864 TI - Analysis of clinical AIDS-free interval after CD4+ cell counts fall below 200 x 10(6) L-1 in Japanese haemophiliacs infected with HIV-1. AB - We analysed the time from the date CD4+ cell counts fell below 200 x 10(6) L-1, defined as ti, to the onset of clinical AIDS, according to the 1987 Centers for Disease Control and Prevention case definition, in 129 Japanese haemophilia patients infected with HIV-1. The cumulative onset of clinical AIDS was analysed by the Kaplan-Meier method and proportional hazard model. Incorporated covariates were age of each patient at time ti, as well as CD4+ and CD8+ cell counts, serum levels of IgG, IgA, IgM, GOT and GPT at ti. The time of antiretroviral treatment initiation was also considered. The 50% AIDS-free interval after ti was 3.00 years (95% confidence interval (CI), range 0.49-5.51) and 1.71 years (95% CI, range 0.66-2.76) for the patients at CDC stage II and stage III, respectively (significantly different, P = 0.0013). Among the patients at CDC stage II at ti, higher levels of IgA were tightly associated with a shorter period from ti to onset of clinical AIDS (P < 0.0001), and relative hazard was 1.35 (95% CI, 1.11 1.64) with increase of IgA level by 1.0 g L-1. Thus there is a broad distribution in the time to onset of clinical AIDS in Japanese haemophiliacs even after CD4+ cell counts fall below 200 x 10(6) L-1. This should be taken into consideration in deciding upon the therapy and care of HIV-1 infected people. PMID- 9873865 TI - Presence of human immunodeficiency virus (HIV) and T-lymphotropic virus type I and II (HTLV-I/II) in a haemophiliac population in Belo Horizonte, Brazil, and correlation with additional serological results. AB - The aim of this study was to determine the prevalence of human immunodeficiency virus type 1 (HIV-1) and human T lymphotropic virus types I and II (HTLV-I/II) infections in 226 haemophiliac patients treated at Fundacao Hemominas in Belo Horizonte, Minas Gerais State, Brazil, and to verify association with other serological results. Patients positive for HTLV-I/II had also a neurological, haematological and ophthalmological evaluation. Fundacao Hemominas offers comprehensive care for all haemophiliac patients in Minas Gerais. Thirty-six (15.9%) of the 226 patients showed reactive results to HIV-1 [ELISA, Abbott, USA, confirmed by Western blot (WB), Cambridge Biotech, USA, and/or immunofluorescence, Fiocruz, Brazil] and 16 (7.1%) had reactive sera to HTLV-I/II (ELISA, Ortho). Eleven of these 16 (4.9%) were positive, 3/16 (1.3%) were indeterminate and 2/16 (0.9%) were negative in the HTLV WB (Cambridge Biotech). Neurological, haematological and ophthalmological examination of 9/16 patients revealed no abnormality suggestive of HTLV disease. Of the 16 patients reactive to HTLV-I/II ELISA test, six (37.5%) were also positive to HIV-1 (chi 2 = 5.92; P = 0.01). Seropositivity for HTLV-I/II and HIV-1 was associated with advancing age and positive results for hepatitis C virus (HCV), Chagas' disease (T. cruzi infection) and syphilis. No association between the presence of HTLV with type and severity of haemophilia and hepatitis B results was detected. The prevalence of antibodies against HIV-1 is approximately three times that of HTLV-I/II and a patient positive for HTLV-I/II had a significantly increased risk of being positive for HIV-1, HCV and T. cruzi. PMID- 9873866 TI - Spinal epidural haematoma in haemophilia A. AB - We report on a 27-year-old male Caucasian with severe haemophilia A who presented with acute onset of neck pain with cervical nerve root irritation, due to a spinal epidural haematoma. His past medical history revealed carrying of a moderate weight as a possible traumatic mechanism. Under immediate factor VIII replacement therapy complete remission of the symptoms was achieved within several days. The diagnosis of spinal epidural haematoma and complete resorption was revealed by initial and follow-up magnetic resonance imaging studies of the cervical spine. Having reviewed the literature on spinal epidural haematoma, we present an overview of the treatment and outcome as regards haemophilia. PMID- 9873867 TI - Continuous infusion of factor IX concentrate to induce immune tolerance in two patients with haemophilia B. AB - Two patients with haemophilia B and high-responding inhibitor to factor IX were subjected to immune tolerance induction according to the Malmo protocol, including high dosage of factor IX, cyclophosphamide and intravenous gammaglobulin. In one of the patients the treatment was preceded by extracorporeal protein A adsorption. Both patients had previously been subjected to immune tolerance induction without success and as an attempt to improve the tolerance induction regimen and lower cost, factor IX was given as continuous infusion, with a dose of around 300 units per kg body weight daily for 3 weeks. The inhibitor level declined in one of the patients but tolerance was not achieved. In the second patient the inhibitor level remained high. Despite the failure of the treatment in these two cases, we propose that the constant antigen load provided by the continuous infusion of modern, safe, purified factor IX concentrate may theoretically be of greater benefit in immune tolerance induction than the varying load resulting from intermittent infusions. Larger study materials are needed to show if this is so. PMID- 9873868 TI - Haemoperitoneum caused by haemorrhagic corpus luteum in a patient with type 3 von Willebrand's disease. AB - In female patients affected by congenital coagulation disorders haemorrhagic corpus luteum must be considered for the differential diagnosis of acute abdomen. We report the complication in a 22-year-old woman with type 3 von Willebrand's disease who presented with abdominal pain and haemorrhagic shock. Along with adequate factor VIII replacement therapy oral contraceptive treatment is the key for the management of this potentially life-threatening complication and the long term prevention of future bleeding episodes of this origin. PMID- 9873869 TI - Menorrhagia in von Willebrand disease successfully treated with single daily dose tranexamic acid. AB - Four cases of menorrhagia in von Willebrand disease were successfully treated with tranexamic acid given in a single daily dose of 4 g for the first 3-5 days of the menstrual cycle. The pathophysiology and pharmacokinetics are discussed. The apparent improved efficacy and acceptability of this new dosing regime have been highlighted. PMID- 9873870 TI - Additional data on the morbidity of central venous access devices in patients with haemophilia. PMID- 9873871 TI - Call for post-licensing human pharmacokinetic studies of administration of recombinant factor IX. PMID- 9873872 TI - Inhibitors occur more frequently in African-American and Latino haemophiliacs. PMID- 9873873 TI - Haemophilia and the mysterious spleen: the story of a hate-love relationship. PMID- 9873874 TI - Gene therapy for haemophilia: how far have we come? PMID- 9873875 TI - Clotting factor concentrates and immune function in haemophilic patients. AB - Patients with haemophilia often exhibit a variety of disturbances in immune function. Although infections with HIV, hepatitis and other viruses no doubt contribute to these abnormalities, chronic exposure to extraneous proteins in clotting factor concentrates (CFCs) may also play a role. Numerous in vitro and ex vivo studies show that protein contaminants--such as immunoglobulins, fibrinogen and fibronectin--can depress various immune function indicators. Generally, such studies show that intermediate-purity CFCs are more inhibitory than very high-purity (e.g. monoclonal-purified) CFCs. In many, but not all, studies, the degree of immunosuppression correlates with the amount of intermediate-purity CFC administered over time. Among various indicators of immune function, CD4+ lymphocyte number is a marker for the progression of HIV infection, and maintenance of CD4+ number is associated with delayed progression. A number of studies suggest that, compared with intermediate-purity CFCs, use of very high-purity CFCs is associated with longer preservation of this class of lymphocytes. However, it remains to be seen whether this translates to improved long-term clinical outcomes. Further research is needed on the impact of CFCs on the immune system. For the time being, however, evidence to date favours the use of very high-purity products because they appear to preserve immune function and reduce the risk of infection with hepatitis and other viruses. PMID- 9873876 TI - Causes of death in Canadians with haemophilia 1980-1995. Association of Hemophilia Clinic Directors of Canada. AB - The life expectancy of individuals with haemophilia was close to that of the general population in the early 1980s. Since then, life expectancy has decreased, due to transfusion-transmitted virus infections. Deaths in individuals with haemophilia were investigated by analysing 2450 records from the Canadian Hemophilia Registry, for the years 1980-1995. Deaths were tabulated by age, year and cause, and compared with that of the Canadian male population by calculating standardized mortality ratios (SMRs). The median life expectancy at 1 year of age was calculated for various subpopulations and the impact of various population characteristics was assessed by survival regression modelling. There were 359 deaths and the annual number of deaths increased significantly after 1986. Risk factors were seropositivity to human immunodeficiency virus (relative risk 16.7, 95% CI 11.1-25.1), severe haemophilia (1.9, 1.3-2.7) and moderate haemophilia (1.8, 1.2-2.6). In HIV antibody negative individuals, the overall death rate was not increased (SMR 0.9, 95% CI 0.7-1.1) and only haemorrhage was significantly increased. In HIV antibody positive individuals, causes of death which were significantly increased were acquired immunodeficiency syndrome, liver failure, haemorrhage, lymphoma, liver cancer, nonspecific infections, and trauma or violence. Deaths due to the acquired immunodeficiency syndrome accounted for only 66% of the excess deaths in individuals who were HIV antibody positive. Life expectancy has markedly decreased since the onset of the HIV epidemic. The impact of HIV is underestimated by considering only deaths due to the acquired immunodeficiency syndrome; other HIV-linked causes need also to be considered. PMID- 9873877 TI - Prednisone and low-dose activated prothrombin complex concentrates for FVIII inhibitor in nonhaemophilic patients. AB - Eight nonhaemophilic patients with factor VIII (FVIII) inhibitors were reported. There was no difference in sex distribution. Median age at diagnosis was 62 years (ranging from 14 to 73 years). No associated disorders were revealed and all the patients were presented with severe muscular or arthral bleeding. Inhibitor titre was measured by the Bethesda method, which were 6.4, 126.0, 155.0, 4.8, 56.0, 13.5, 35.0 and 150.0 BU mL-1, respectively, at diagnosis. FVIII:C levels were less than 1 U dL-1 in seven patients and less than 2 U dL-1 in one patient. The median vWF:Ag level was 210% (ranging from 80% to 340%). All the patients had good response to activated prothrombin complex concentrates for acute bleeding episodes and prednisone for inhibitor elimination. Inhibitors completely eliminated in seven patients within a follow-up duration over 1 year, and one patient died of intracranial haemorrhage when her inhibitor titre decreased to 4.5 BU mL-1 and FVIII:C increased to 21 U dL-1. PMID- 9873878 TI - Increased CD4-positive monocytes in HIV-infected haemophilic patients. AB - The monocyte-macrophage system is known to play a central role in HIV infection, and expression of CD4 on the surface of monocytes/macrophages is important, since this molecule is a key factor for the entrance of HIV into susceptible cells. In this paper we evaluated the expression of CD4 in monocytes of haemophilic patients (He) who had been infected with HIV (HIV + He) through transfusion of contaminated plasma concentrates. Thirty seropositive patients (HIV + He), 10 seronegative He patients (HIV-He) and 20 voluntary normal blood donors were studied. Phenotypic evaluation of monocytes was performed by flow cytometry of peripheral blood stained with anti-CD45, -CD3, -CD4 and -CD14 monoclonal antibodies. The percentage of CD4 monocytes was increased in all HIV+ patients groups, but it was highest in those belonging to Groups III and IV A of the CDC classification. Furthermore, the median of fluorescence intensity of CD4+ monocytes from individual patients was shifted to the right, indicating expression of increased numbers of CD4 molecules on the cell membrane of monocytes. This could in turn favour HIV infection and viral persistence, facilitating in vivo dissemination of the virus. PMID- 9873880 TI - Laboratory performance of haemophilia centres in developing countries: 3 years' experience of the World Federation of Hemophilia External Quality Assessment Scheme. AB - A World Federation of Hemophilia External Quality Assessment Scheme has been established to promote high standards of laboratory performance in haemophilia centres worldwide. Results from 21 International Haemophilia Training Centres (IHTCs) provide target values for the prothrombin time (PT), activated partial thromboplastin time (APTT), factor VIII:C, IX:C and von Willebrand factor (VWF) assays, against which the performance of Haemophilia Centres in developing countries (HCs) can be assessed. Eight surveys were distributed over a 3-year period between 1994 and 1997. A higher proportion of HCs failed to identify an abnormal PT or APTT in samples from donors with mild deficiencies of the extrinsic and intrinsic systems, respectively. For factor VIII:C and IX:C assays, agreement between HC results was consistently poorer than between IHTCs. However, improvement in between-centre agreement could be seen for two samples distributed on more than one occasion. A minority of HCs perform assays for VWF, and a questionnaire revealed equipment and reagent costs as limiting the range of assays which could be carried out in several centres. However, agreement was in some cases better between those HCs that did perform VWF assays than between IHTCs. The problems of screening test sensitivity and between-centre agreement for factor assays need to be addressed, together with the limitations which prevent HCs from performing a full range of tests in the diagnosis and treatment of bleeding disorders. PMID- 9873879 TI - Laboratory and clinical markers of HIV infection in a national haemophilia cohort treated with recombinant factor VIII concentrate. The Association of Hemophilia Clinic Directors of Canada. AB - Over an interval of approximately six months beginning in October 1993, most haemophilia A patients in Canada were switched from a plasma-derived intermediate purity factor VIII concentrate (i.p. VIII) to a recombinant factor VIII (rVIII). In order to determine the consequence of this change in therapy on progression of HIV infection, we gathered surveillance data on clinical status and CD4 and CD8 cell counts in those patients who were HIV seropositive at the time of switching concentrates. Data were recorded at the time of switchover, annually for 2 years thereafter, and retrospectively at a point 1 year prior to the switch. CD4 cells fell significantly over the study period. Multiple direct comparisons revealed that this decline was restricted to the time intervals which included the final year in which patients received intermediate-purity factor VIII concentrate (i.p. VIII). In the 2 year interval in which rVIII was used exclusively, there was a nonsignificant fall in CD4 cells. Changes in CD4 cells did not correlate with the intensity of exposure to either i.p.VIII or rVIII. CD8 cells did not fall significantly over the study period. There was no obvious reduction in the incidence of death or clinical progression over the 2 years in which rVIII was used. However, we are hopeful that the stabilizing trend in CD4 cell counts which followed the introduction of rVIII will be predictive of corresponding clinical stabilization over the coming years. PMID- 9873881 TI - Total knee and hip arthroplasty in haemophilic patients. AB - We evaluated the long-term results of three total hip and nine total knee arthroplasties in nine haemophilic patients with disabling pain and end-stage arthropathy. These patients have been followed over a period of 2-12 years (mean 6.9). One patient had a post-operative haematoma, which was evacuated; two patients required manipulation of the knee because of a limited range of motion. There were no infections. At follow-up, all but two patients were completely free of pain; two patients had occasional pain. The functional improvement was impressive with an average increase in the knee score from 37 to 80 points post operatively. The hip score increased from 36 to 85 points. The range of motion was increased in seven joints, unchanged in two joints and decreased in three. One total hip arthroplasty was revised after 9 years for aseptic loosening. One total knee demonstrated nonprogressive radiographic evidence of aseptic loosening. Median consumption of coagulation factor concentrate in severe haemophiliacs decreased from 47.00 U yr-1 (range 16.000-144.000) before surgery, to 25.000 U yr-1 (range 18.000-132.000) at 2 years after surgery. CONCLUSION: Hip and knee arthroplasty is a valuable option in symptomatic haemophilic patients with disabling arthropathy, in order to obtain pain relief and functional improvement. It is associated with a low rate of complications and may reduce the need for substitution of factor VIII and IX. PMID- 9873882 TI - An unusual haemarthrosis in an HIV-seronegative haemophilia A patient. AB - We report the case of a severe haemophilia A patient with an anti-factor VIII antibody who presented with a thigh haematoma and 1 year later with an elbow haemarthrosis infected by Salmonella enteritidis. These two infections were treated by antibiotics. The probable origin of these infections seems to be an anal fistula. The occurrence of a septic arthritis due to Salmonella is rare, and to our knowledge has never been reported in HIV-negative haemophilic patients. The differential diagnosis of haemarthrosis and septic arthritis in a haemophilic patient is also discussed. PMID- 9873883 TI - Development of a factor VIII inhibitor in a newborn haemophiliac. AB - A major problem in the treatment of haemophilia A is the development of inhibitors (antibodies) against factor VIII. We report the case of a newborn male with no family history of haemophilia who developed an intracerebral haemorrhage. On day 10 post-delivery severe haemophilia A was diagnosed and treatment with recombinant FVIII (rFVIII) concentrate was started. Seventy-two hours later the presence of inhibitors was suspected because high doses of rFVIII were required to maintain therapeutic FVIII plasma levels. Days after, the inhibitor was detected. The quick detection of the inhibitor in this newborn haemophiliac allowed us to start the immunotolerance early, without interruption in the administration of rFVIII. PMID- 9873884 TI - Development of a subdural vein thrombosis following aggressive factor VII replacement for postnatal intracranial haemorrhage in a homozygous factor VII deficient infant. AB - Congenital factor VII deficiency is a rare (1:500,000) autosomally recessive coagulopathy with variable expression and high penetration. In infants the most devastating presentation is that of intracranial haemorrhage. An infant is described with severe factor VII deficiency who developed postnatal intracranial haemorrhage. The baby was treated with factor VII concentrate (ImmunoA.G., Vienna, Austria). Three weeks after the haemorrhage he developed a dural venous sinus thrombosis. Although factor VII-deficient patients may need treatment with factor VII concentrate, this needs to be carefully monitored because of the thrombotic risk. PMID- 9873885 TI - Recombinant factor VIIa in continuous infusion during central line insertion in a child with factor VIII high-titre inhibitor. AB - Recombinant factor VIIa (rFVIIa) is a recently added new tool for the treatment of haemophilia patients with inhibitors. A major drawback in the use of rFVIIa is its short half-life, which necessitates frequent bolus injections. Thus the use of rFVIIa in continuous infusion appears to be a good alternative. We describe the use of rFVIIa, administered by continuous infusion with a minipump during the insertion of a central venous catheter in a child with a high-titre factor VIII inhibitor. rFVIIa was administered as an intravenous bolus (90 micrograms kg-1 [4.5 kIU kg-1]), 1 h prior to central line insertion, after which the continuous infusion was immediately started for 5 days. The infusion rate was based on the clearance obtained from a previous pharmacokinetic study. Effective haemostasis and normal healing of surgical incisions were achieved after central line insertion. No local thrombophlebitis nor evidence of generalized activation of the coagulation cascade was observed. Single-dose pharmacokinetic parameter values were clearance (Cl) 34.6 mL h-1 kg-1, volume of distribution (Vd) 40.6 mL kg-1 and mean residence time (MRT) 1.17 h. The recovery was 2.27% U-1 kg-1. rFVIIa showed a monophasic decay. Cl during continuous infusion was 23.4 +/- 6.9 mL h-1 kg-1. The administration of rFVIIa by continuous infusion is effective, safe and more convenient when compared to other clotting factors. Moreover, continuous infusion provides significant economic savings (77% decrease in rFVIIa requirements). PMID- 9873886 TI - Does protease inhibitor treatment induce increased bleeding tendency in haemophilia? PMID- 9873888 TI - Witnessing medical history: an interview with Dr Rosemary Biggs. Interview by Christine Lee and Charles Rizza. PMID- 9873887 TI - Recurrent haemoperitoneum in a female patient with type III von Willebrand's disease responded to administration of oral contraceptive. PMID- 9873889 TI - Molecular genetics of coagulation factor VIII gene and haemophilia A. PMID- 9873891 TI - Meeting the needs of haemophilic children in developing countries. PMID- 9873890 TI - Laboratory diagnosis of hereditary bleeding disorders: external quality assessment. AB - A World Federation of Hemophilia external quality assessment scheme has been established to promote high standards of laboratory performance in haemophilia centres world-wide. Results from 22 International Haemophilia Training Centres (IHTCs) provide target values for the prothrombin time (PT), activated partial thromboplastin time (APTT), factor VIII:C, IX:C and von Willebrand factor assays, against which the performance of haemophilia centres (HCs) in developing countries can be assessed. A higher proportion of HCs failed to identify an abnormal PT or APTT in samples from donors with mild deficiencies of the extrinsic and intrinsic systems, respectively. For factor VIII:C and IX:C assays, agreement between HC results was consistently poorer than between IHTCs. However, improvement in between-centre agreement could be seen for two samples distributed on more than one occasion. A minority of HCs perform assays for von Willebrand factor, and a questionnaire revealed equipment and reagent costs as limiting the range of assays which could be carried out in several centres. However, agreement was in some cases better between those HCs that did perform VWF assays, than between IHTCs. The problems of screening test sensitivity, and between-centre agreement for factor assays need to be addressed, together with the limitations which prevent HCs from performing a full range of tests in the diagnosis and treatment of bleeding disorders. PMID- 9873892 TI - Genetic services available for counselling and prenatal diagnosis of haemophilia. PMID- 9873893 TI - Maintenance of musculoskeletal function in people with haemophilia. AB - Bleeding affecting muscles and joints is the most common manifestation of haemophilia. In the long term, prevention of musculoskeletal abnormality in haemophilic patients is cost effective. Early diagnosis and proper treatment is fundamental to prevent disability. Prevention can be divided into secondary and tertiary levels. Secondary prevention concentrates on how to handle a haemophilic child, and on the education of the family so that they develop a safe and healthy lifestyle and awareness of the importance of early diagnosis and treatment. Tertiary prevention deals with minimizing the musculoskeletal consequences of muscle and joint bleeding. Coagulation factor replacement, appropriate care of the bleeding site and early physical therapy helps to prevent complications and yields the best results. Recurrent episodes of joint bleeding can lead to chronic synovitis. Initially synovitis demands conservative treatment. However, conservative methods are often not successful. Chronic synovitis is the most important cause of joint degeneration and the aim of rehabilitation is to achieve functional status using special casting techniques, orthoses or preventative surgery. Surgical, radioisotope or chemical synovectomy may be used. Technical requirements and availability of haemostatic cover limit the use of surgical and radioisotope synovectomy. The author has found chemical synovectomy with rifampicin to be an easier and more effective procedure than surgery. Problems related to joint deformities, muscle atrophy and chronicle pain demand comprehensive care. They are the main factors affecting the maintenance of musculoskeletal function and the social integration of people with haemophilia. PMID- 9873894 TI - Delivery of haemophilia care in the developing world. PMID- 9873895 TI - Strategic planning for health care in developing countries. PMID- 9873896 TI - The transfer of technology from the developed to the developing world. AB - The treatment of haemophilia in both the developed and developing world countries need not be complicated. The person with haemophilia can be treated and managed by common sense and the application of basic knowledge and understanding of the physiology and pathology. Emphasis needs to be placed on training people rather than on technology in the areas of clinical medicine, laboratory medicine and blood transfusion medicine. Within these specialties, there are a number of practical aspects in which a critical degree of expertise is required. Such expertise may not be available in the developing country and hence there is a very justifiable reason for the exchange of knowledge, technology and goodwill. PMID- 9873897 TI - Public health and international health-care development for persons with haemophilia: operation improvement and operation access. PMID- 9873898 TI - World Federation of Hemophilia developing world programmes. PMID- 9873899 TI - WHO/WFH recommendations, June 1997. Hemophilia care in developing countries. PMID- 9873900 TI - A symbolic environment for visualizing activated foci in functional neuroimaging datasets. AB - This paper presents a symbolic visualization environment known as the Corner Cube environment, which was developed to facilitate rapid examination and comparison of activated foci defined by analyses of functional neuroimaging datasets. We have performed a comparative evaluation of this environment against maximum intensity projection and 'gallery of slices' displays, and the results suggest that the Corner Cube environment has definite advantages over both conventional display techniques. We conclude that the Corner Cube is an effective tool for summarizing the spatial characteristics of activated foci within an easily understood visual context and is especially useful for displaying the similarities and differences in functional neuroimaging datasets. PMID- 9873901 TI - The detection and significance of subtle changes in mixed-signal brain lesions by serial MRI scan matching and spatial normalization. AB - The purpose of this work is to detect and assess the significance of subtle signal changes in mixed-signal lesions based on serial MRI scan matching. Pairs of serially acquired T1-weighted volume MR images from 20 normal controls and seven patients with epilepsy were matched and difference images obtained. The precision and consistency of the registration were evaluated. The Gaussian noise level in the difference images was determined automatically. A structured difference filter was then used to segment structured (changed) voxels from the Gaussian noise. In the controls, the structured difference images were normalized into Talairach space, resulting in a structured noise map. The significance of changes in patients was assessed by spatial normalization and comparison with the structured noise map. The precision and consistency of the co-registration were < or = 0.06 mm with a registration success rate of 100%. The Gaussian noise level in the difference images was in the range 3.0-6.9. In the controls, an average of 1.6% of the brain voxels were classified as structured. Sine-based registration resulted in a reduction of < 1% in the amount of structure compared to linear interpolation. The structured noise map in controls showed high noise density in areas affected by image artefacts. We show examples of significant changes found in lesions which had been reported as unchanged on visual inspection. A novel quantitative approach has been presented for the detection and quantification of subtle signal changes in lesions. This method is of potential clinical value in the non-invasive characterization of signal change and biological behaviour of neoplastic lesions. PMID- 9873902 TI - Image matching as a diffusion process: an analogy with Maxwell's demons. AB - In this paper, we present the concept of diffusing models to perform image-to image matching. Having two images to match, the main idea is to consider the objects boundaries in one image as semi-permeable membranes and to let the other image, considered as a deformable grid model, diffuse through these interfaces, by the action of effectors situated within the membranes. We illustrate this concept by an analogy with Maxwell's demons. We show that this concept relates to more traditional ones, based on attraction, with an intermediate step being optical flow techniques. We use the concept of diffusing models to derive three different non-rigid matching algorithms, one using all the intensity levels in the static image, one using only contour points, and a last one operating on already segmented images. Finally, we present results with synthesized deformations and real medical images, with applications to heart motion tracking and three-dimensional inter-patients matching. PMID- 9873903 TI - On-line analysis of echocardiographic image sequences. AB - This paper presents a semiautomatic system for the interactive analysis of echocardiographic image sequences, able to provide useful information to cardiologists. The proposed approach combines well known techniques for the detection of left ventricular boundaries with the computation of optical flow. The initial detection of the cavity contour is based on an improved balloon model, with automatic tuning of parameters and optional model-based constraints. The computation of optical flow is performed with a fast correlation technique and the contour tracking is obtained combining motion information provided by the optical flow with model-based constraints and/or a snake-based regularization. The system is able to follow the motion of the ventricular boundaries precisely, to provide several quantitative features of the heartbeat and a dynamic representation of systolic and diastolic motion. Preliminary experimental results are presented and commented on with particular attention to their clinical relevance. Furthermore a distributed implementation of the system suitable for remote examination analysis has been realized and applied to echo images of the carotid artery. PMID- 9873904 TI - Motion and deformation tracking for short-axis echo-planar myocardial perfusion imaging. AB - The assessment of regional myocardial perfusion during the first-pass of a contrast agent bolus requires tracking of the signal time course for each myocardial segment so that a detailed perfusion map can be derived. To obtain such a map in practice, however, is not trivial because deformation of the shape of the myocardium and respiratory-induced motion render a major difficulty in this process. This study describes an automated approach for motion and deformation tracking of functional myocardial perfusion images. The effectiveness of the described method has been evaluated using a numerical phantom and results are compared with those from existing techniques which use deformable models. Preliminary results from applying our approach to 20 patients are discussed and compared with those from SPECT studies. PMID- 9873905 TI - Three-dimensional spatial compounding of ultrasound images. AB - One of the most promising applications of 3-D ultrasound lies in the visualization and volume estimation of internal 3-D structures. Unfortunately, the quality of the ultrasound data can be severely degraded by artefacts and speckle, making automatic analysis of the 3-D data sets very difficult. In this paper we investigate the use of 3-D spatial compounding to reduce speckle. We develop a new statistical theory to predict the improvement in signal-to-noise ratio with increased levels of compounding, and verify the predictions empirically. We also investigate how registration errors can affect automatic volume estimation of structures within the compounded 3-D data set. Having established the need to correct these errors, we present a novel reconstruction algorithm which uses landmarks to register each B-scan accurately as it is inserted into the voxel array. In a series of in vitro and in vivo trials, we demonstrate that 3-D spatial compounding is very effective for improving the signal-to-noise ratio, but correction of registration errors is essential. PMID- 9873906 TI - Demonstration of accuracy and clinical versatility of mutual information for automatic multimodality image fusion using affine and thin-plate spline warped geometric deformations. AB - This paper applies and evaluates an automatic mutual information-based registration algorithm across a broad spectrum of multimodal volume data sets. The algorithm requires little or no pre-processing, minimal user input and easily implements either affine, i.e. linear or thin-plate spline (TPS) warped registrations. We have evaluated the algorithm in phantom studies as well as in selected cases where few other algorithms could perform as well, if at all, to demonstrate the value of this new method. Pairs of multimodal gray-scale volume data sets were registered by iteratively changing registration parameters to maximize mutual information. Quantitative registration errors were assessed in registrations of a thorax phantom using PET/CT and in the National Library of Medicine's Visible Male using MRI T2-/T1-weighted acquisitions. Registrations of diverse clinical data sets were demonstrated including rotate-translate mapping of PET/MRI brain scans with significant missing data, full affine mapping of thoracic PET/CT and rotate-translate mapping of abdominal SPECT/CT. A five-point thin-plate spline (TPS) warped registration of thoracic PET/CT is also demonstrated. The registration algorithm converged in times ranging between 3.5 and 31 min for affine clinical registrations and 57 min for TPS warping. Mean error vector lengths for rotate-translate registrations were measured to be subvoxel in phantoms. More importantly the rotate-translate algorithm performs well even with missing data. The demonstrated clinical fusions are qualitatively excellent at all levels. We conclude that such automatic, rapid, robust algorithms significantly increase the likelihood that multimodality registrations will be routinely used to aid clinical diagnoses and post-therapeutic assessment in the near future. PMID- 9873907 TI - Analysis of dynamic MR breast images using a model of contrast enhancement. AB - We describe a model of dynamic contrast enhancement in breast MRI designed to aid the radiologist in cases for which X-ray mammography is ineffective. The breasts are segmented from the image slices by a dynamic programming algorithm after morphological opening. A pharmacokinetic model has been derived to fit the rise in intensities after injection of a contrast agent, in a way that facilitates investigation of the effects of different models of bolus injection. The pharmacokinetic model is used in a modified Horn-Schunck algorithm to correct for motion effects during the seven minute acquisition period. The results show significant localization of tumours and enable discrimination of cancerous tissue. In particular, we illustrate the approach with an image that shows a carcinoma, whose appearance and localization are greatly improved by the registration algorithm. PMID- 9873908 TI - Landmark methods for forms without landmarks: morphometrics of group differences in outline shape. AB - Morphometrics, a new branch of statistics, combines tools from geometry, computer graphics and biometrics in techniques for the multivariate analysis of biological shape variation. Although medical image analysts typically prefer to represent scenes by way of curving outlines or surfaces, the most recent developments in this associated statistical methodology have emphasized the domain of landmark data: size and shape of configurations of discrete, named points in two or three dimensions. This paper introduces a combination of Procrustes analysis and thin plate splines, the two most powerful tools of landmark-based morphometrics, for multivariate analysis of curving outlines in samples of biomedical images. The thin-plate spline is used to assign point-to-point correspondences, called semi landmarks, between curves of similar but variable shape, while the standard algorithm for Procrustes shape averages and shape coordinates is altered to accord with the ways in which semi-landmarks formally differ from more traditional landmark loci. Subsequent multivariate statistics and visualization proceed mainly as in the landmark-based methods. The combination provides a range of complementary filters, from high pass to low pass, for effects on outline shape in grouped studies. The low-pass version is based on the spectrum of the spline, the high pass, on a familiar special case of Procrustes analysis. This hybrid method is demonstrated in a comparison of the shape of the corpus callosum from mid-sagittal sections of MRI of 25 human brains, 12 normal and 13 with schizophrenia. PMID- 9873909 TI - Estimation of three-dimensional cardiac velocity fields: assessment of a differential method and application to three-dimensional CT data. AB - We have investigated an optical flow method for the estimation of the three dimensional endocardial wall motion from high-resolution X-ray CT data. This method was originally proposed by Song and Leahy. It is based on the optical flow, the divergence-free and the smoothness constraints. Due to the characteristics of the imaging modality, we studied the restriction of this approach to the boundary of the left ventricular (LV) cavity. The behaviour of the method is quantified through simulations approximating the overall motion of the LV cavity through an affine transform involving a dilation and a rotation. The method implies the choice of three parameters weighting the constraints. The results show a weak dependence of the velocity field on the weighting of the optical flow constraint. The accuracy of the method relies more heavily on the relative weighting of the smoothness and divergence-free constraints. In our experiments, the best results were obtained for a largely predominant divergence free constraint. The results also show that the accuracy of the method is reasonable for low values of the rotation angle (minimum mean error of 1.1 voxel for 5 degrees). This is compatible with values reported in other studies for the overall rotation of the LV. We provide a qualitative description of the results obtained in vivo on a canine heart by visualizing the distribution of the estimated velocity vector magnitudes over the endocardial surface. These results (evolution of the field over time, maximum velocities) are in agreement with the known physiological behaviour of the heart. PMID- 9873910 TI - Segmentation of the visible human for high-quality volume-based visualization. AB - This article describes a combination of interactive classification and super sampling visualization algorithms that greatly enhances the realism of 3-D reconstructions of the Visible Human data sets. Objects are classified on the basis of ellipsoidal regions in RGB space. The ellipsoids are used for super sampling in the visualization process. PMID- 9873911 TI - Detection, visualization and animation of abnormal anatomic structure with a deformable probabilistic brain atlas based on random vector field transformations. AB - This paper describes the design, implementation and preliminary results of a technique for creating a comprehensive probabilistic atlas of the human brain based on high-dimensional vector field transformations. The goal of the atlas is to detect and quantify distributed patterns of deviation from normal anatomy, in a 3-D brain image from any given subject. The algorithm analyzes a reference population of normal scans and automatically generates color-coded probability maps of the anatomy of new subjects. Given a 3-D brain image of a new subject, the algorithm calculates a set of high-dimensional volumetric maps (with typically 384(2) x 256 x 3 approximately 10(8) degrees of freedom) elastically deforming this scan into structural correspondence with other scans, selected one by one from an anatomic image database. The family of volumetric warps thus constructed encodes statistical properties and directional biases of local anatomical variation throughout the architecture of the brain. A probability space of random transformations, based on the theory of anisotropic Gaussian random fields, is then developed to reflect the observed variability in stereotaxic space of the points whose correspondences are found by the warping algorithm. A complete system of 384(2) x 256 probability density functions is computed, yielding confidence limits in stereotaxic space for the location of every point represented in the 3-D image lattice of the new subject's brain. Color-coded probability maps are generated, densely defined throughout the anatomy of the new subject. These indicate locally the probability of each anatomic point being unusually situated, given the distributions of corresponding points in the scans of normal subjects. 3-D MRI and high-resolution cryosection volumes are analyzed from subjects with metastatic tumors and Alzheimer's disease. Gradual variations and continuous deformations of the underlying anatomy are simulated and their dynamic effects on regional probability maps are animated in video format (on the accompanying CD-ROM). Applications of the deformable probabilistic atlas include the transfer of multi-subject 3-D functional, vascular and histologic maps onto a single anatomic template, the mapping of 3-D atlases onto the scans of new subjects, and the rapid detection, quantification and mapping of local shape changes in 3-D medical images in disease and during normal or abnormal growth and development. PMID- 9873912 TI - Finding parametric representations of the cortical sulci using an active contour model. AB - The cortical sulci are brain structures resembling thin convoluted ribbons embedded in three dimensions. The importance of the sulci lies primarily in their relation to the cytoarchitectonic and functional organization of the underlying cortex and in their utilization as features in non-rigid registration methods. This paper presents a methodology for extracting parametric representations of the cerebral sulcus from magnetic resonance images. The proposed methodology is based on deformable models utilizing characteristics of the cortical shape. Specifically, a parametric representation of a sulcus is determined by the motion of an active contour along the medial surface of the corresponding cortical fold. The active contour is initialized along the outer boundary of the brain and deforms toward the deep root of a sulcus under the influence of an external force field, restricting it to lie along the medial surface of the particular cortical fold. A parametric representation of the medial surface of the sulcus is obtained as the active contour traverses the sulcus. Based on the first fundamental form of this representation, the location and degree of an interruption of a sulcus can be readily quantified; based on its second fundamental form, shape properties of the sulcus can be determined. This methodology is tested on magnetic resonance images and it is applied to three medical imaging problems: quantitative morphological analysis of the central sulcus; mapping of functional activation along the primary motor cortex and non-rigid registration of brain images. PMID- 9873913 TI - The Brain Bench: virtual tools for stereotactic frame neurosurgery. AB - We present a suite of neurosurgery supporting tools developed around (i) the Virtual Workbench, a productive environment for the control of 3-D data, in which delicate work can be performed for hours on end without strain, and (ii) the Electronic Brain Atlas, integrating the major print brain atlases in day-to-day clinical use. We describe in detail the Brain Bench, a surgical planning system for stereotactic frame neurosurgery. Its objective is to prepare faster plans; have a better and more accurate choice of target points; improve the avoidance of sensitive structures; have fewer sub-optimal frame attachments and speedier, more effective planning and training. If validated by a clinical study now under way, this will improve medical efficacy and reduce costs. PMID- 9873914 TI - Interactive live-wire boundary extraction. AB - Live-wire segmentation is a new interactive tool for efficient, accurate and reproducible boundary extraction which requires minimal user input with a mouse. Optimal boundaries are computed and selected at interactive rates as the user moves the mouse starting from a manually specified seed point. When the mouse position comes into the proximity of an object edge, a 'live-wire' boundary snaps to, and wraps around the object of interest. The input of a new seed point 'freezes' the selected boundary segment and the process is repeated until the boundary is complete. Two novel enhancements to the basic live-wire methodology include boundary cooling and on-the-fly training. Data-driven boundary cooling generates seed points automatically and further reduces user input. On-the-fly training adapts the dynamic boundary to edges of current interest. Using the live wire technique, boundaries are extracted in one-fifth of the time required for manual tracing, but with 4.4 times greater accuracy and 4.8 times greater reproducibility. In particular, interobserver reproducibility using the live-wire tool is 3.8 times greater than intraobserver reproducibility using manual tracing. PMID- 9873915 TI - Fast numerical algorithms for fitting multiresolution hybrid shape models to brain MRI. AB - In this paper, we present new and fast numerical algorithms for shape recovery from brain MRI using multiresolution hybrid shape models. In this modeling framework, shapes are represented by a core rigid shape characterized by a superquadric function and a superimposed displacement function which is characterized by a membrane spline discretized using the finite-element method. Fitting the model to brain MRI data is cast as an energy minimization problem which is solved numerically. We present three new computational methods for model fitting to data. These methods involve novel mathematical derivations that lead to efficient numerical solutions of the model fitting problem. The first method involves using the nonlinear conjugate gradient technique with a diagonal Hessian preconditioner. The second method involves the nonlinear conjugate gradient in the outer loop for solving global parameters of the model and a preconditioned conjugate gradient scheme for solving the local parameters of the model. The third method involves the nonlinear conjugate gradient in the outer loop for solving the global parameters and a combination of the Schur complement formula and the alternating direction-implicit method for solving the local parameters of the model. We demonstrate the efficiency of our model fitting methods via experiments on several MR brain scans. PMID- 9873916 TI - Automated lumen definition from 30 MHz intravascular ultrasound images. AB - One prerequisite for standard clinical use of intravascular ultrasound imaging is rapid evaluation of the data. The main quantities to be extracted from the data are the size and the shape of the lumen. Until now, no accurate, robust and reproducible method to obtain the lumen boundaries from intravascular ultrasound images has been described. In this study, 21 different (semi-)automated binary segmentation methods for determining the lumen are compared with manual segmentation to find an alternative for the laborious and subjective procedure of manual editing. After a preprocessing step in which the catheter area is filled with lumen-like grey values, all approaches consist of two steps: (i) smoothing the images with different filtering methods and (ii) extracting the lumen by an object definition method. The combination of different filtering methods and object definition methods results in a total of 21 methods and 80 experiments. The results are compared with a reference image, obtained from manual editing, by use of four different quality parameters--two based on squared distances and two based on Mahalanobis distances. The evaluation has been carried out on 15 images, of which seven are obtained before balloon dilation and eight after balloon dilation. While for the post-dilation images no definite conclusions can be drawn, an automated contour model applied to images smoothed with a large kernel appears to be a good alternative to manual contouring. For pre-dilation images a fully automated active contour model, initialized by thresholding, preceded by filtering with a small-scale median filter is the best alternative for manual delineation. The results of this method are even better than manual segmentation, i.e. they are consistently closer to the reference image than the average distance of all individual manual segmentations. PMID- 9873917 TI - A robust point-matching algorithm for autoradiograph alignment. AB - We present a novel method for the geometric alignment of autoradiographs of the brain. The method is based on finding the spatial mapping and the one-to-one correspondences (or homologies) between point features extracted from the images and rejecting non-homologies as outliers. In this way, we attempt to account for the local, natural and artifactual differences between the autoradiograph slices. We have used the resulting automated algorithm on a set of left prefrontal cortex autoradiograph slices, specifically demonstrated its ability to perform point outlier rejection, validated its robustness property using synthetically generated spatial mappings and provided an anecdotal visual comparison with the well-known iterated closest-point (ICP) algorithm. Visualization of a stack of aligned left prefrontal cortex autoradiograph slices is also provided. PMID- 9873918 TI - A representation for mammographic image processing. AB - Mammographic image analysis is typically performed using standard, general purpose algorithms. We note the dangers of this approach and show that an alternative physics-model-based approach can be developed to calibrate the mammographic imaging process. This enables us to obtain, at each pixel, a quantitative measure of the breast tissue. The measure we use is h(int) and this represents the thickness of 'interesting' (non-fat) tissue between the pixel and the X-ray source. The thicknesses over the image constitute what we term the h(int) representation, and it can most usefully be regarded as a surface that conveys information about the anatomy of the breast. The representation allows image enhancement through removing the effects of degrading factors, and also effective image normalization since all changes in the image due to variations in the imaging conditions have been removed. Furthermore, the h(int) representation gives us a basis upon which to build object models and to reason about breast anatomy. We use this ability to choose features that are robust to breast compression and variations in breast composition. In this paper we describe the h(int) representation, show how it can be computed, and then illustrate how it can be applied to a variety of mammographic image processing tasks. The breast thickness turns out to be a key parameter in the computation of h(int), but it is not normally recorded. We show how the breast thickness can be estimated from an image, and examine the sensitivity of h(int) to this estimate. We then show how we can simulate any projective X-ray examination and can simulate the appearance of anatomical structures within the breast. We follow this with a comparison between the h(int) representation and conventional representations with respect to invariance to imaging conditions and the surrounding tissue. Initial results indicate that image analysis is far more robust when specific consideration is taken of the imaging process and the h(int) representation is used. PMID- 9873919 TI - Segmentation of 2-D and 3-D objects from MRI volume data using constrained elastic deformations of flexible Fourier contour and surface models. AB - This paper describes a new model-based segmentation technique combining desirable properties of physical models (snakes), shape representation by Fourier parametrization, and modelling of natural shape variability. Flexible parametric shape models are represented by a parameter vector describing the mean contour and by a set of eigenmodes of the parameters characterizing the shape variation. Usually the segmentation process is divided into an initial placement of the mean model and an elastic deformation restricted to the model variability. This, however leads to a separation of biological variation due to a global similarity transform from small-scale shape changes originating from elastic deformations of the normalized model contours only. The performance can be considerably improved by building shape models normalized with respect to a small set of stable landmarks (AC-PC in our application) and by explaining the remaining variability among a series of images with the model flexibility. This way the image interpretation is solved by a new coarse-to-fine segmentation procedure based on the set of deformation eigenmodes, making a separate initialization step unnecessary. Although straightforward, the extension to 3-D is severely impeded by difficulties arising during the generation of a proper surface parametrization for arbitrary objects with spherical topology. We apply a newly developed surface parametrization which achieves a uniform mapping between object surface and parameter space. The 3-D procedure is demonstrated by segmenting deep structures of the human brain from MR volume data. PMID- 9873920 TI - Multi-modal volume registration by maximization of mutual information. AB - A new information-theoretic approach is presented for finding the registration of volumetric medical images of differing modalities. Registration is achieved by adjustment of the relative position and orientation until the mutual information between the images is maximized. In our derivation of the registration procedure, few assumptions are made about the nature of the imaging process. As a result the algorithms are quite general and can foreseeably be used with a wide variety of imaging devices. This approach works directly with image data; no pre-processing or segmentation is required. This technique is, however, more flexible and robust than other intensity-based techniques like correlation. Additionally, it has an efficient implementation that is based on stochastic approximation. Experiments are presented that demonstrate the approach registering magnetic resonance (MR) images with computed tomography (CT) images, and with positron-emission tomography (PET) images. Surgical applications of the registration method are described. PMID- 9873921 TI - Analysis of left ventricular wall motion based on volumetric deformable models and MRI-SPAMM. AB - We present a new approach for the analysis of the left ventricular shape and motion based on the development of a new class of volumetric deformable models. We estimate the deformation and complex motion of the left ventricle (LV) in terms of a few parameters that are functions and whose values vary locally across the LV. These parameters capture the radial and longitudinal contraction, the axial twisting, and the long-axis deformation. Using Lagrangian dynamics and finite-element theory, we convert these volumetric primitives into dynamic models that deform due to forces exerted by the datapoints. We present experiments where we used magnetic tagging (MRI-SPAMM) to acquire datapoints from the LV during systole. By applying our method to MRI-SPAMM datapoints, we were able to characterize the 3-D shape and motion of the LV both locally and globally, in a clinically useful way. In addition, based on the model parameters we were able to extract quantitative differences between normal and abnormal hearts and visualize them in a way that is useful to physicians. PMID- 9873922 TI - Development and preliminary evaluation of VISLAN, a surgical planning and guidance system using intra-operative video imaging. AB - VISLAN is an integrated neurosurgical planning and guidance system. New segmentation and rendering techniques have been incorporated. A stereo video system is used intra-operatively and fulfils four roles. First, the video display is overlaid with graphical outlines showing the position of the planned craniotomy or the target (enhanced reality displays). Second, a skin surface patch is reconstructed from the stereo video images using patterned light (mean errors of surface point location are < 0.15 mm). Third, a freely mobile, hand held localizer is tracked in real time (position errors are < 0.5 mm and with improved calibration < 0.2 mm), with its position superimposed on the pre operative patient representation to assist surgical guidance. Fourth, markers fixed to the skull bone next to the cranial opening are used to detect intra operative movement and to update registration. Initial results from phantom experiments show an overall system accuracy of better than 0.9 mm for intra operative localization of features defined in pre-operative images. The prototype system has been tested during six neurosurgical operations with very good results. PMID- 9873923 TI - Deformable models in medical image analysis: a survey. AB - This article surveys deformable models, a promising and vigorously researched computer-assisted medical image analysis technique. Among model-based techniques, deformable models offer a unique and powerful approach to image analysis that combines geometry, physics and approximation theory. They have proven to be effective in segmenting, matching and tracking anatomic structures by exploiting (bottom-up) constraints derived from the image data together with (top-down) a priori knowledge about the location, size and shape of these structures. Deformable models are capable of accommodating the significant variability of biological structures over time and across different individuals. Furthermore, they support highly intuitive interaction mechanisms that, when necessary, allow medical scientists and practitioners to bring their expertise to bear on the model-based image interpretation task. This article reviews the rapidly expanding body of work on the development and application of deformable models to problems of fundamental importance in medical image analysis, including segmentation, shape representation, matching and motion tracking. PMID- 9873924 TI - Segmentation of brain tissue from magnetic resonance images. AB - Segmentation of medical imagery is a challenging problem due to the complexity of the images, as well as to the absence of models of the anatomy that fully capture the possible deformations in each structure. The brain is a particularly complex structure, and its segmentation is an important step for many problems, including studies in temporal change detection of morphology, and 3-D visualizations for surgical planning. We present a method for segmentation of brain tissue from magnetic resonance images that is a combination of three existing techniques from the computer vision literature: expectation/maximization segmentation, binary mathematical morphology, and active contour models. Each of these techniques has been customized for the problem of brain tissue segmentation such that the resultant method is more robust than its components. Finally, we present the results of a parallel implementation of this method on IBM's supercomputer Power Visualization System for a database of 20 brain scans each with 256 x 256 x 124 voxels and validate those results against segmentations generated by neuroanatomy experts. PMID- 9873925 TI - Tracking and motion analysis of the left ventricle with deformable superquadrics. AB - We present a new approach to analyse the deformation of the left ventricle of the heart based on a parametric model that gives a compact representation of a set of points in a 3-D image. We present a strategy for tracking surfaces in a sequence of 3-D cardiac images. Following tracking, we then infer quantitative parameters which characterize: left ventricle motion, volume of left ventricle, ejection fraction, amplitude and twist component of cardiac motion. We explain the computation of these parameters using our model. Experimental results are shown in time sequences of two modalities of medical images, nuclear medicine and X-ray computed tomography (CT). Video sequences presenting these results are on the CD ROM. PMID- 9873926 TI - Comparison of edge-based and ridge-based registration of CT and MR brain images. AB - In modern medicine, several different imaging techniques are frequently employed in the study of a single patient. This is useful, since different images show complementary information on the functionality and/or structure of the anatomy examined. This very difference between modalities, however, complicates the problem of proper registration of the images involved, and rules out the most basic approaches--like direct grey value correlation--to achieve registration. The observation that some common structures will always exist is supportive of the statement that registration may be feasible using edges or ridges present in the images. The existence of such structures defined in the binary sense is questionable, however, and their extraction from images requires a segmentation by definition. In this paper we propose to use fuzzy edgeness and ridgeness images, thus avoiding the need for segmentation and using more of the available information from the original images. We will show that such fuzzy images can be used to achieve accurate registration. Several ridgeness and edgeness computing operators were compared. The best registration results were obtained using a gradient magnitude operator. PMID- 9873927 TI - Automated 3-D registration of MR and CT images of the head. AB - This paper discusses the application of voxel similarity measures in the automated registration of clinically acquired MR and CT data of the head. We describe a novel single-start multi-resolution approach to the optimization of these measures, and the issues involved in applying this to data having a range of different fields of view and sampling resolution. We compare four proposed measures of voxel similarity using the same optimization scheme when presented with 10 pairs of images with a range of initial misregistrations. The registration estimates are compared with those provided by manual point-based registration and evaluated by visual inspection to give an assessment of the robustness and accuracy of the different measures. One full-volume CT image set is used to investigate the performance of each measure when used to align truncated images from different regions in the head. The soft tissue correlation and mutual information measures were found to provide the most robust measures of misregistration, providing results comparable to or better than those from manual point-based registration for all but the most truncated image volumes. PMID- 9873928 TI - Outlines of the 42 years adenovirus research in the Institute of Microbiology, Semmelweis Medical University. PMID- 9873929 TI - Interrelationship of viruses and cells. The cytopathic property of adenoviruses. AB - The application of in vitro cell cultures in virus research has a great importance in studies on the interaction between virus and cell. A great number of the viruses causes characteristic degeneration in the infected cells, well visible in the light-microscope, and due to their cytopathic effect the existence of viruses became directly detectable. Based on characteristic changes even the serological type of virus can be determined in some cases. Thus the multilateral examination of the interaction between virus and host cell has immense importance both in virus research and in therapy. PMID- 9873930 TI - The haemagglutination spectra of adenoviruses and factors influencing the haemagglutinating activity. PMID- 9873931 TI - Serological and virus isolation experiments on the incidence of adenoviruses in Hungary. PMID- 9873932 TI - Characterization of the main protein components of adenovirus virion and its possible use in laboratory diagnostics. PMID- 9873933 TI - Pathomechanism of adenovirus infections. PMID- 9873934 TI - Studies on tumourous and other urogenital patients with respect to antigens of oncogenic adenovirus. AB - The possible connection of viruses with tumours was investigated by serologic examinations. Concerning the presence of antibodies against adenoviruses, especially those against the early non-virion antigens of oncogenic adenovirus type 12, approximately 4000 tests were made with sera of 446 urogenital patients with and without tumours and 70 ones with internal diseases. It was found by complement fixation tests that antibodies against nonvirion antigens of adenoviruses were present in 53% of urogenital patients suffering from malignant tumours and prostatic hypertrophy, in 18% of non-tumourous urological patients and in 4% of patients with internal diseases, respectively. The results suggest that adenoviruses may play a role in tumourous diseases of the urogenital organs. PMID- 9873935 TI - Molecular biological characterization of adenovirus DNA. AB - The agarose gel electrophoresis of DNA, the ethidium bromide fluorescence detection of DNA fragments and restriction endonucleases were discovered at the end of the '60s. The methodological progress enabled institutions equipped with less sophisticated technology to achieve also unique experimental and scientific results in the field of viral DNA research. The team working on virus DNA within the adenovirus research group has constructed several new restriction endonuclease maps of the genomes of human and animal adenoviruses; contributed to the methodology of the determination of specific endonuclease sites, and genome polarity; discovered new restriction endonucleases, adenovirus subtypes, new empty capsid, and genome subpopulations; participated in cooperations leading to novel, although hypothetical approaches in AIDS therapy, taxonomic definition of viruses, and evolutionary origins of adenovirus replication and encapsidation strategy. PMID- 9873936 TI - Molecular structure of the two-dimensional hexon crystalline array and of adenovirus capsid. PMID- 9873937 TI - Intertype specific epitope structure of adenovirus hexon. PMID- 9873938 TI - Outlines of the immunological research in the Institute of Microbiology; Semmelweis Medical University. AB - The working group on immunology under the leadership of Pal Foldes began its activity with poliovirus studies during the severe epidemics of 1957. It was he who first in Hungary isolated poliovirus strain from patients [1]. His colleague was Ilona, Szeri who had gained her first experiences in virology at Jozsef Sinkovics's virus laboratory. Then Zsuzsanna Banos and Piroska Anderlik joined them and became permanent members of the working group on immunology. Since 1965 with the leadership of Ilona Szeri, they have been conducting basic researches into immunology for over three decades at the Institute, with a wide sphere of collaborators. Research has been intended to acquire more thorough and precise knowledge of the role in immunobiology of the thymus and lymphoid system and of pathogenesis of the wasting syndrome as well as of interactions of the microorganisms and the organism. The most significant results are going to be summed up in the following. PMID- 9873939 TI - Biological effects of microorganisms and their substances in mice. AB - In our experiments we evidence from several aspects that the normal microbial flora has a permanent and life-long immune modulating role in conventional organisms and a stimulating effect both on specific and non-specific defense. However, in case of artificial interventions (stress, drugs) affecting the organism, existence of the normal flora may have an adverse effect on it (endotoxin effect, bacterial translocation). The immunomodulants show a stimulating effect mainly in organisms with undeveloped immune system, and their effects are independent from the presence or absence of the microbial flora. With ageing, effect of immunomodulants can change and become indifferent or even suppressive. Dose-dependence of stimulating or suppressing effect of immunomodulants may be related to their non-immunological effects (endotoxin effect, bacterial translocation). Finally, on the basis of our experiments, we consider the Gf mouse suitable for examining the effect of a given agent in the practice, on the one hand, and for observing the host organism's reactions, free from the influence of the normal microbial flora, on the other. Along with the known physiological and pathological events, our results draw also attention to as distant fields as drug sensitivity, drug interactions influencing drug sensitivity. The authors put emphasis on importance of germfree environment during immunosuppressive treatments in humans and when making special examinations under experimental conditions. PMID- 9873940 TI - Modulation of the cellular immune response to intracerebral lymphocytic choriomeningitis virus (LCMV) infection in mice. AB - Certain viruses do not kill the cells they infect. The immunological response of the host in such situations may be involved in the development of pathologic changes and clinical illness. Pioneering work by Rowe has shown that death associated with acute LCMV infection in the mouse is resulted from the immune response. Many investigators using a variety of techniques including neonatal thymectomy, irradiation, or treatment of adult mice with antilymphoid drugs or antithymocyte sera have confirmed and extended Rowe's observations. The study of LCMV and the disease it causes in its natural murine host has provided the initial findings that open new fields in viral immunobiology, viral immunopathology, and cell-cell recognition. PMID- 9873941 TI - Correlation between thymic function and skeletal growth. PMID- 9873942 TI - Potential role of microbiological agents in sudden infant death syndrome. AB - The potential role of microbiological agents was investigated in 10 cases of Sudden Infant Death Syndrome in Budapest between September 1996 and December 1997. Autopsy, histological examination and microbiological tests were performed on samples of blood, cerebrospinal fluid, pharyngeal and bronchial samples from infants under six months died suddenly, without previous diseases. The multifactorial pathomechanism of SIDS was suggested by detection of Parainfluenza Type virus antigen, isolation of toxin producing Staphylococcus aureus, Enterobacteriaceae and Candida albicans strains in large number of more samples of the same infant. PMID- 9873943 TI - The origin and evolution of viruses (a review). AB - Viroids and prions might have existed early at the border of inanimate and living worlds. Most extant viruses can be characterized as derivatives of ancestors originating from episomal elements of prokaryotes (DNA phages) and later from eukaryotes. Retroviruses very likely originated from cellular retrotransposons. Retrograde evolution of some large viruses from obligatory intracellular bacteria is possible but the ontogenesis of extant bacteria does not include a viral form of existence (the filterable L forms are not viruses) and well-defined viruses do not regenerate back into vegetative bacterial forms. Biologists experimenting with the evolution of prokaryotic and eukaryotic ancient cells cannot ignore the earliest appearance of viruses within or outside the living matter. Viruses participated in and gave direction to the evolution and natural selection by coexisting with uni- and multicellular organisms for billions of years. The coevolution of viruses and their host cells is characterized by incessant attacks and counterattacks through gene rearrangements and mutations (induced in the virus by an immunological counterattack of the host or by transgression of species barriers by the virus) and recombinations. Recombinations occurred between viral and viral or viral and host genes. Acts of "molecular piracy" as practiced by ancient viruses endowed the virus with the expression of several host genes for the advantage of the virus in its replicative cycle and host-to host spread. Probably the first immortalized and malignantly transformed cells were induced by viruses as viruses evolved anti-apoptotic measures. While infected cells resort to apoptotic death before the assembly of a new viral progeny, prominent are the anti-apoptotic measures viruses evolved in order to assure the completion of their full replicative cycle. Further, viruses may escape neutralization by host antibodies and may survive a counterattack by the host's T cells directed at virally infected cells of its own. Viruses may induce a form of tolerance and coexist with their host without inducing disease. Persistent and apparently or deceivingly apathogenic or even attenuated viral "quasi-species" populations may contain individual particles that regain virulence due to recombinations and/or gene rearrangements, especially when transgressing species barriers. Xenotropic viruses of animals may replicate in human cells and vice versa confounding experiments with xenotransplants or with use of veterinary viral vaccines for the treatment of human diseases. PMID- 9873944 TI - New assays for the quality control of live oral poliovirus vaccine. AB - The strains of all three types of poliovirus used in the production of live oral poliomyelitis vaccine have been shown to yield vaccines that are both immunogenic and highly attenuated when administered orally to susceptible children and adults. Experience obtained for close to four decades with the vaccines prepared from these strains indicates that laboratory and animal tests described in the World Health Organization's (WHO) Requirements for Poliomyelitis Vaccines (Oral) do ensure the consistency of virus characteristics during vaccine production. Major advances in our understanding of the molecular basis of attenuation and reversion of polioviruses resulted in the development of a new generation of tests. These include an alternative in vivo neurovirulence test in transgenic mice that express the human poliovirus receptor and a new in vitro assay (mutant analysis by polymerase chain reaction and restriction enzyme cleavage, shortly MAPREC) that assess the consistency of vaccine production at a molecular level. A WHO collaborative study was initiated in 1993 with the objective of assessing transgenic mice as potential models for evaluation of the neurovirulence of type 3 vaccines. The results of the study showed that there is a very good correlation between the TgPVR21 mouse assay and the monkey neurovirulence test. Further studies are in progress to develop a statistical model for making regulatory decisions on accepting or rejecting batches of type 3 vaccine. A WHO collaborative study was initiated in 1991 to evaluate the MAPREC assay for type 3 vaccines. The results of this study showed that the MAPREC test was a sensitive, robust and standardized molecular assay suitable for process development for new manufacturers and for monitoring the consistency of existing vaccine production. Screening single virus harvests with MAPREC before pooling them into monovalent bulk vaccine will also improve the quality of the final product. PMID- 9873945 TI - Advance in bacterial typing methods (a review). AB - We give a short review about the two main groups of bacterial typing methods for epidemiological purposes: the phenotypic and genotypic methods. The advantage of the phenotypic methods is their feasibility for the less equipped laboratories, their disadvantages are the variability of the phenotypic properties, and their high labor requirement. The advantages of the genotypic typing methods are the higher discriminatory power and applicability of the same method for different species of bacteria. The disadvantage of these methods is that they are expensive and labour consuming. Beside the short description, we show the results of different authors obtained by the use of the molecular methods in the epidemiological practice and we call the attention to the insufficiency concerning the stability. PMID- 9873946 TI - The acute phase reaction syndrome: the acute phase reactants (a review). AB - The acute phase reaction of the organism, which determines the response that follows the injury of tissues and organs and helps the survival of the individual is reviewed. The main participants, the induction of the reaction and the regulatory mechanisms are shortly discussed, with special respect to the prominent role of interleukin-6. The relationship between the production and synthesis of the acute phase reactants under physiological and pathophysiological conditions is analyzed. The particular functions of orosomucoid and alpha 2HS glycoprotein are summarized. Finally, the common biological actions of IL-6 and the acute phase reactants are mentioned. PMID- 9873947 TI - Isolation of human alpha 2HS-glycoprotein synthesized by Sf9 cells. AB - The human alpha 2HS-glycoprotein is a negative acute phase protein synthesized by hepatocytes. Because of its fragility and the difficulty of its purification, we used recombinant DNA techniques to produce the protein in order to investigate its biological effects. The cDNA coding for the whole alpha 2HS-glycoprotein from human liver library had been cloned into the baculovirus expression vector system using the pVL 1392 transfer vector and Sf9 cells. The recombinant protein was synthesized in the Sf9 cells and was isolated on a hydroxiapatite column from the culture medium. Western blot analysis indicated that the cells synthesized large quantities of the recombinant human protein. The molecular mass of the recombinant AHSG was the same as that of the protein in human plasma but was slightly lower than that of AHSG in the cell culture supernatants of HepG2 and higher than that of AHSG from Hep3B cell cultures, respectively. PMID- 9873948 TI - First isolation of enteric adenoviruses type 41 from children with acute gastroenteritis in Hungary. AB - In a retrospective study, 122 stool specimens, collected 1989-1990, from children with gastroenteritis were found to contain adenoviruses. Twenty-nine adenoviruses of 50 randomly selected adenovirus-positive fecal samples were successfully propagated in Graham 293 cells and were typed by DNA restriction enzyme analysis with SmaI. Six strains were typed as adenovirus 41 (Ad41), and 23 strains as non enteric adenovirus. All six adenoviruses type 41 had the same DNA pattern identical with that of the variant Ad41/D12. Ad40 viruses were not detected. Enteric Ad41 infections were observed only in males and were found throughout the year. This is the first report on isolation and typing of enteric adenoviruses 41 in Hungary. PMID- 9873949 TI - Investigation of the presence of different broad-spectrum beta-lactamases among clinical isolates of Enterobacteriacae. AB - Chromosomal or plasmid-encoded beta-lactamases are the most frequent causes of resistance to broad-spectrum beta-lactam antibiotics in clinical isolates of Gram negative bacteria. Different screening methods can be used for their detection during routine laboratory work, while molecular biological methods may help in the detection of the genetic background of the phenotypic resistance. Clinical isolates of Klebsiella pneumoniae (170) and Enterobacter cloaceae (82) were obtained from different parts of Hungary, whereas those of Serratia marcescens (15) were isolated in our Department from a nosocomial outbreak. Disk diffusion and the Etest were used to screen inducible Class C beta-lactamase and plasmid mediated extended spectrum beta-lactamases (ESBLs) among clinical isolates of Enterobacteriaceae. Single-strand conformation polymorphism (SSCP) analysis of the PCR products obtained after using SHV-specific primers revealed the presence of SHV-2 and SHV-5 ESBL among 170 K. pneumoniae strains in 12 and 3 cases, respectively. The results of the screening methods and the PCR-SSCP analysis suggested that 14 of the 15 S. marcescens strains not only produced the Class C, inducible chromosomal beta-lactamase, but also acquired a plasmid-mediated SHV-2 type ESBL. One strain isolated from the environment during the outbreak was genetically related to the other isolates, as demonstrated by the different typing methods, but it did not produce ESBL. The in vivo transfer of SHV-2 gene was assumed from an SHV-2 positive K. pneumoniae strain present in the same ward, in the same patient and at the same time. A very high prevalence of the stable derepressed mutants of E. cloaceae was confirmed among the Hungarian isolates. Seventy seven per cent of the strains produced high amounts of beta-lactamase without induction being responsible for their resistance to third-generation cephalosporins. Nineteen per cent of the strains were inducible when cefoxitin or imipenem was used, as confirmed by direct measurement of the MICs with the Etest. PMID- 9873951 TI - Gastrointestinal hormones as potential adjuvant treatment of exocrine pancreatic adenocarcinoma. AB - CONCLUSION: Gastrointestinal hormones and their antagonists can alter the growth of pancreatic adenocarcinoma in vitro and in vivo. The potential clinical benefit of this approach deserves further study. BACKGROUND: Epithelial cell growth is normally under hormonal control. Hormones also affect the growth of many epithelial cancers, and this fact is used to modify tumor growth. Pancreatic epithelial cell growth is under the influence of gastrointestinal hormones. This article reviews experiments designed to determine the effect of gastrointestinal hormones on the growth of pancreatic adenocarcinoma. METHODS: Eighty-eight articles were identified from a Medline search using the terms pancreatic adenocarcinoma and the individual names of gastrointestinal hormones. The experimental design and results of these studies are reviewed. RESULTS: In general, somatostatin, vasoactive intestinal polypeptide, pancreatic polypeptide, and pancreastatin inhibit pancreatic adenocarcinoma growth. Cholecystokinin, secretin, bombesin, gastrin, EGF, TGF-alpha, insulin, and IGF-1 have a growth promoting effect. PMID- 9873952 TI - Measurement of pS2 protein in pancreatic cyst fluids. Evidence for a potential role of pS2 protein in the pathogenesis of mucinous cystic tumors. AB - CONCLUSION: Elevated levels of the growth factor pS2 protein in the cyst fluids of mucinous cystic tumors correlate with earlier observations using immunohistochemical techniques showing that pS2 protein is expressed by these tumors. The markedly elevated levels of pS2 protein compared to normal plasma values suggest that this growth factor may be important in the pathogenesis of pancreatic mucinous cystic tumors. BACKGROUND: Cystic lesions of the pancreas include inflammatory pseudocysts, serous cystadenomas, and mucinous cystic tumors, some of which are malignant. Previous studies using immunohistochemical techniques have shown that virtually all pancreatic mucinous tumors express pS2 protein. pS2 protein is a growth factor that is believed to be important in the normal process of inflammation and repair. We measured pS2 protein and other growth factors in pancreatic cyst fluids to assess their potential pathophysiologic and diagnostic significance. METHODS: Levels of pS2 protein were measured in 54 pancreatic cyst fluids by radioimmunoassay. The growth factors, epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), and insulin-like growth factors I and II (IGF-I, IGF-II) were measured in 22 cyst fluids using commercial immunoassays. RESULTS: Mucinous cysts exhibited significantly higher levels of cyst fluid pS2 protein than nonmucinous lesions, including pseudocysts and serous cystadenomas (median: 78,303 pg/mL; range: 218 361,176 pg/mL vs median: 886 pg/mL; range: 0-14,206 pg/mL; p = 0.0001). The level of pS2 in mucinous tumors was markedly higher than plasma values (median: 392 pg/mL). Levels of pS2 protein in malignant mucinous lesions tended to be higher than those in benign mucinous cysts, but this difference was not statistically significant (median: 88,817 vs 64,350 pg/mL; p = 0.159). Levels of other growth factors, including EGF, TGF-alpha, IGF-I, and IGF-II, did not discriminate among the different cyst types, and the values were within normal plasma ranges. PMID- 9873950 TI - Clinical islet cell transplantation. Are we there yet? AB - Diabetes mellitus is perhaps the most devastating chronic disease of all time. A brief history of the evolution of treatment modalities is provided, culminating in the rationale for the physiologic replacement of a functioning beta-cell mass by transplantation. Vascularized pancreas transplantation is discussed briefly as an introduction to the transplantation of the isolated islet. A detailed review of the current state of human islet transplantation for the cure of diabetes is then described. Finally, areas for future development are highlighted. PMID- 9873953 TI - Difference in microbes contaminating pancreatic necrosis in biliary and alcoholic pancreatitis. AB - CONCLUSION: There are differences in the microbiology of infected pancreatic necrosis in alcoholic and biliary pancreatitis. One possible explanation may be different routes of contamination. BACKGROUND: Infection is a severe complication in acute pancreatitis. Bacteria are found in 40-70% of all patients suffering from necrotizing pancreatitis. We investigated whether there were any differences in microbes isolated from pancreatic necrosis in biliary and alcoholic pancreatitis. METHODS: Microbiological tests were conducted on necrosis taken at the operation for pancreatitis with the etiology of (group A) alcoholic pancreatitis (n = 47) and (group B) biliary pancreatitis (n = 23). Patients with simultaneous cholecystitis were excluded. The time from the first symptoms to the operation or the extent of necrosis did not differ between the groups. RESULTS: Microbes were isolated more often in the cultures from group B than group A (17/23 = 74% vs 15/47 = 32%, p = 0.001). The most common were Gram-positive bacteria in group A and Gram-negative bacteria in group B. From the first week, from the onset of symptoms to the operation. Gram-negative bacteria were isolated significantly more often in the cultures from group B patients than from group A patients (8/10 = 80% vs 1/5 = 20%, p = 0.04). In multivariate analysis, we found that biliary pancreatitis was an independent risk factor (adds ratio 5.5, 95% confidence interval [CI] 0.59-52.10) of contamination of necrosis with Gram negative bacteria. PMID- 9873954 TI - Trypsinogen activation and glutathione content are linked to pancreatic injury in models of biliary acute pancreatitis. AB - CONCLUSION: In models of biliary acute pancreatitis, which might resemble the situation in humans, premature activation of trypsinogen inside the pancreas ("autodigestion") occurs and is correlated with the extent of ductal and parenchymal injury. It is accompanied by a critical spending of protease inhibitors and glutathione, compromising important acinar cell defense and maintenance mechanisms. BACKGROUND: Premature activation of pancreatic digestive enzymes and profound changes of levels of certain biochemical compounds have been implicated in the pathophysiology of acute pancreatitis. Hitherto, little information on their role in biliary acute pancreatitis has been available. METHODS: Three types of injury to the pancreaticobiliary duct system of various severity were induced in rats--ligation of the common bile-pancreatic duct, retrograde infusion of electrolyte, or retrograde infusion of taurocholate solution--and were compared to sham-operated animals. Trypsin, trypsin inhibitory capacity (TIC), reduced glutathione (GSH), and other compounds were measured in pancreatic tissue. Histopathology, as well as serum amylase, lipase, and gamma glutamyl transferase (gamma GT) were assessed. RESULTS: Histopathology and elevated activity of gamma GT in the serum revealed increasing severity of pancreatic injury from sham operation through retrograde duct infusion with taurocholate. GSH was diminished even in macroscopically normal-appearing tissue, but significantly lower in altered (hemorrhagic)-looking sections. Conversely, tissue levels of trypsin were significantly increased. TIC was elevated only in the duct obstruction model, whereas it was reduced in the retrograde duct infusion models. PMID- 9873955 TI - Beneficial effect of octreotide treatment in acute pancreatitis in rats. AB - CONCLUSIONS: Octreotide treatment contributes to the regulation of tumor necrosis factor (TNF) production in sodium taurocholate-induced acute necrotizing pancreatitis in rats. Owing to its complex effect, octreotide can partially ameliorate the deleterious consequences of acute necrotizing pancreatitis. Elevated TNF and interleukin-6 (IL-6) levels in the peritoneal fluid may be considered a consequence of the activation of peritoneal macrophages. BACKGROUND: The effects of octreotide on exocrine pancreatic function have been investigated in numerous studies, but little attention has been paid to its influence on cytokine production in acute pancreatitis. METHODS: Acute pancreatitis was induced by the retrograde injection of taurocholic acid into the pancreatic duct in male Wistar rats. Serum amylase activity, wet pancreatic weight/body weight (pw/bw) ratio, and TNF and IL-6 levels were measured. Four micrograms/kg of octreotide was administered subcutaneously at the time of induction of pancreatitis and 24 or 48 h later. Rats were sacrificed 6, 24, 48, or 72 h after the operation. RESULTS: The serum amylase level and pancreatic weight to body weight ratio were decreased significantly in the octreotide-treated group. The serum TNF level was decreased significantly in the octreotide-treated group as compared with the control group at 6, 24, and 48 h (0.6 +/- 1.5, 2.0 +/- 3.3, and 0 vs 50 +/- 15.5, 37.5 +/- 18.4, and 13.1 +/- 12.5 U/mL, respectively). The ascites TNF level was decreased to 0 in the octreotide-treated group and was elevated in the control group at 72 h (28.0 +/- 49.0 U/mL). IL-6 production in ascites was extremely high in both groups at 6 h (80,000 +/- 43,817 pg/mL and 58,500 +/- 33,335 pg/mL), but the difference was not significant. PMID- 9873957 TI - Papillary-cystic neoplasm of the pancreas. A sex-steroid dependent tumor. AB - CONCLUSION: The rate of growth of a papillary-cystic tumor of the pancreas seemed to be enhanced by the concurrence of pregnancy. Progesterone receptors in the tumor were demonstrated by immunohistochemistry and by molecular biology methods. BACKGROUND: Papillary cystic tumor of the pancreas is extremely rare, occurring predominantly young females. Owing to the low frequency of the tumor, there is scarce information about the conditions that promote tumor growth. METHODS: In this report, we present the temporal association between very rapid growth of a papillary-cystic neoplasm and pregnancy. Clinicopathological, immunohistochemical, and molecular biology analyses were performed. RESULTS: A 21 yr-old woman was admitted because of recurrent epigastric abdominal pain associated with episodes of nausea and vomiting, and a history of an abdominal tumor of about 50 mm near the head of the pancreas, detected by ultrasound. On admission the patient had a flat, nontender abdomen without palpable masses. Laboratory analysis including hematologic and hepatic tests were strictly normal; only CA 19-9 (42 U/mL, normal 37 U/mL) was above normal values. One week after admission, an abdominal computerized axial tomography (CAT) scan revealed an 81.6 mm cystic mass localized in the head of the pancreas, and 1 wk later, in a laparotomy, a papillary-cystic neoplasm of 120 mm, limited to the head of the pancreas, was found. Three months later, in a routine follow-up visit, an 18-wk pregnancy was clinically diagnosed and confirmed by ultrasound exploration. The pregnancy continued without complications, and a normal male infant (3.7 kg) was born at 39 wk of gestation, by vaginal delivery. Eighteen months after tumor resection, the patient was asymptomatic and her child was in good health. We propose that progesterone affects tumor growth. PMID- 9873956 TI - The effect of intermittent injections of CCK-8S and the CCK-A receptor antagonist devazepide on cell proliferation in exocrine rat pancreas. AB - CONCLUSION: Intermittent injections of sulfated cholecystokinin-8 (CCK-8S) or devazepide caused long-lasting effects on cell proliferation in exocrine pancreas in contrast to continuous infusion. The acinar cells responded to both treatments with changes in the labeling index (LI) during the whole study period. When studying the influence of stimulation and inhibition of the CCK-A receptor on cell proliferation in the exocrine pancreas, not only are the drugs and doses of importance but also the mode of administration. BACKGROUND: Continuous infusion of CCK-8S or the CCK-A receptor antagonist devazepide induces transient changes in acinar cell proliferation in rat pancreas. The aim of the present experiments was to study whether intermittent administration of CCK-8S or devazepide prevents receptor desensitization and thereby affects exocrine pancreatic cell proliferation persistently. METHODS: Male Sprague-Dawley rats were injected subcutaneously twice daily with CCK-8S (6 micrograms), devazepide (240 micrograms) or bovine serum albumin (BSA). The rats were sacrificed after 18 and 36 h and 3 and 7 d. One hour before sacrifice, the rats were injected intraperitoneally with 1 mCi/kg of tritiated thymidine. The pancreatic weight and the contents of water, protein, and DNA were determined. The LI (number of labeled cells/100 cells) of exocrine pancreatic cells was determined microscopically after autoradiography. RESULTS: The concentration of plasma CCK was slightly increased by devazepide, but the increase was more pronounced by CCK 8S. The pancreatic wet weight was transiently increased 18 h after the start of CCK-8S injections (+14%), whereas devazepide caused a reduction after 7 d (-22%). The protein content was uninfluenced and the DNA content was decreased at 36 h with either treatment. CCK-8S increased the LI in acinar and centroacinar cells throughout the study period, but the ductal cell LI was increased only after 18 and 36 h. Injection of devazepide was followed by decreased LI of acinar cells throughout the study period. Also, the centroacinar and ductal cell LI decreased initially but returned to control values after 7 d. PMID- 9873958 TI - Clinicopathologic differentiation of atrophy of the pancreatic body and tail aplasia. AB - CONCLUSION: Congenital aplasia of the body and tail of the pancreas is derived from a defect of the dorsal pancreatic anlage and should not be considered a type of acquired atrophy of these structures. BACKGROUND: Congenital aplasia of the body and tail of the pancreas radiologically mimics acquired atrophy of the pancreatic body and tail. METHODS: Two patients with radiologically identified aplasia of the body and tail of the pancreas were studied clinicopathologically. RESULTS: An 82-yr-old man was diagnosed radiologically as having both carcinoma of the head of the pancreas and aplasia of the body and tail of the pancreas and underwent pancreatoduodenectomy. Pathologically the carcinoma was distributed in the anterosuperior part of the head of the pancreas, and spread into the duct of Santorini and intraductally to a portion of the main pancreatic duct beyond the junction of the ducts of Santorini and Wirsung. Consequently, obstructive pancreatitis of the body and tail of the pancreas developed, resulting in marked atrophy that mimicked aplasia of the body and tail of the pancreas. A 74-yr-old woman was diagnosed radiologically as having two carcinomas, one of the gallbladder and one of the stomach, and aplasia of the body and tail of the pancreas. During surgery, suspected parenchymal disappearance and fatty replacement in the body and tail of the pancreas were noted. Histologic examination of biopsy specimens from the body portion revealed atrophic pancreatic tissue surrounded by fat. Therefore, these patients had atrophy of the pancreatic body and tail. PMID- 9873959 TI - Ischemic stricture of the small intestine associated with acute pancreatitis. AB - A 37-yr-old man underwent an open drainage operation for severe acute pancreatitis and received respiratory ventilation support for 4 mo because of respiratory failure based on disseminated intravascular coagulation (DIC) and septic shock. Under intensive care, he sometimes had bloody diarrhea for about 6 wk. Colonoscopic findings suggested that the bleeding had derived from the small intestine. The patient then gradually recovered from acute pancreatitis and was discharged from the hospital. Thereafter, he suffered relapses of ileus and his symptoms progressively worsened. The patient underwent a second operation about 2 yr after the onset of acute pancreatitis. At celiotomy, multiple stenoses of the distal ileum measuring about 60 cm in length were found and the segment was resected. The resected specimen demonstrated six separate circumferential strictures and shallow ulcerations. Histologically, multiple ulcerations were restricted to the mucosa and were accompanied by marked submucosal edema and fibrosis. The mucosa between the ulcers revealed chronic regenerative changes: intimal thickening of small mesenteric arteries causing luminal narrowing and organized thrombosis in small mesenteric veins. Therefore, these were considered to be a series of segmental ischemic lesions. Note that delayed ischemic stricture of the small intestine may occur as a chronic complication of acute pancreatitis. PMID- 9873960 TI - Stellate ganglion block does not change the tympanic membrane temperatures of either block or unblock sides in male volunteers. AB - STUDY OBJECTIVES: To test the effect of stellate ganglion block on tympanic membrane temperatures. DESIGN: Prospective, observational study. SETTING: Department of Anesthesia, Yamanashi Medical University, Yamanashi, Japan. SUBJECTS: 7 healthy male volunteers. INTERVENTIONS: Stellate ganglion block, 6 ml of 1% mepivacaine hydrochloride, was administered. MEASUREMENTS AND MAIN RESULTS: Application of temperature probes (right and left tympanic membranes, eight adhesive right skin-surface temperatures at the chest, upper arm, digital fingertip, lateral calf, thigh, and great toe) was followed by a 30-minute control period. Temperatures were recorded every 15 minutes for 30 minutes before stellate ganglion block and every 5 minutes for 40 minutes after the block. The right tympanic membrane temperature increased slightly but not significantly compared with the left tympanic membrane temperature 10 minutes after stellate ganglion block and subsequently. Right skin-surface temperature gradients at the upper limb decreased slightly but not significantly after stellate ganglion block. Similarly, right skin-surface temperature gradients at the lower limb decreased slightly but not significantly after stellate ganglion block. CONCLUSIONS: Stellate ganglion block does not change the tympanic membrane temperatures of either block or unblock sides. PMID- 9873962 TI - Onset and recovery of neuromuscular blockade after two doses of rocuronium in children. AB - STUDY OBJECTIVE: To determine if 450 micrograms/kg (1.5 times the ED95) of rocuronium would result in a comparable onset with a shorter duration of action when compared with 600 micrograms/kg (2 times the ED95). DESIGN: Randomized, single-blind study. SETTING: Teaching hospital. PATIENTS: 85 ASA physical status I and II children ages 2 through 12, undergoing elective surgery with an inhalation induction using halothane. INTERVENTIONS: Group 1 received 600 micrograms/kg rocuronium, and Group 2 received 450 micrograms/kg rocuronium. MEASUREMENTS AND MAIN RESULTS: The two groups were compared using a Student's t test, with p < 0.05 significant. The time of onset, or time to 95% suppression of neuromuscular twitch with standard errors, was 140 +/- 13 seconds (range 46 to 365 sec) in Group 1 and 148 +/- 13 seconds (range 82 to 345 sec) in Group 2 (NS = not significant). The times to 25% return of twitch from baseline (T25) in Groups 1 and 2 were 28 +/- 1.5 minutes (range 14 to 45 min) and 26 +/- 1.6 minutes (range 10 to 55 min), respectively (NS). The differences between these two doses in onset of, and recovery from, block were not found to be statistically significant. The results, however, excluded 5% of the higher dose group and 31% of the lower dose group who did not achieve 95% suppression of twitch. Time to maximal suppression of neuromuscular blockade, however, was not statistically significant for the 85 patients with a time of 270 +/- 28 seconds (range 91 to 605 sec) with a mean maximal suppression of 98.7% in Group 1 and 313 +/- 25 seconds (range 91 to 899 sec) with a mean maximal suppression of 93.1% in Group 2. CONCLUSION: The two doses of rocuronium did not differ statistically in onset or duration. Rocuronium at 600 micrograms/kg offers more reliability than 450 micrograms/kg in achieving adequate muscle relaxation, and the lower dose may result in a significantly large number of patients who may have inadequate intubating conditions. PMID- 9873961 TI - The induction, maintenance, and recovery characteristics of spinal versus general anesthesia in elderly patients. AB - STUDY OBJECTIVE: To compare the induction and recovery profiles of three combinations of general anesthesia when used as an alternative to spinal anesthesia for elderly patients. DESIGN: Randomized, prospective, open-label study. SETTING: Large referral hospital. PATIENTS: 100 [ASA physical status I, II, and III] patients over 60 years of age undergoing brief transurethral surgery. INTERVENTIONS: In Groups Propofol-Propofol (P-P), Propofol-Isoflurane (P I), and Propofol-Desflurane (P-D), anesthesia was induced with fentanyl (1 to 2 micrograms/kg i.v.) and propofol (1.0 to 2.0 mg/kg i.v.) and maintained with 70% nitrous oxide in oxygen and either a propofol infusion (75 to 150 micrograms/kg/min) or isoflurane (end-tidal 0.7% to 1.2%) or desflurane (end tidal 1% to 4%), respectively. After induction, a laryngeal mask airway was placed and spontaneous ventilation was maintained. In Group Spinal (S), 1.5 ml 4% lidocaine (60 mg), in an equal volume of 10% dextrose, was administered intrathecally. MEASUREMENTS AND MAIN RESULTS: Induction and recovery characteristics were compared. Induction with propofol was technically easier and significantly (medp < 0.0001) faster (4.6 +/- 1.7 min, 4.7 +/- 2.2 min, and 3.8 +/- 1.4 min for Groups P-P, P-I, and P-D, respectively) than induction of spinal anesthesia (9.3 +/- 3.4 min). During the induction period, mean arterial blood pressure and heart rate were significantly higher in Group S. Emergence, extubation, and orientation times were similar among the general anesthesia treatment groups. In Group S, patient-generated pain scores were lower (p < 0.05) and recovery room admission longer (p < 0.001). Time to return to baseline digit symbol substitution test (DSST) scores was marginally improved in Groups P-P and P-D when compared to Group P-I. Postoperative nausea, sleepiness, anxiety, and coordination were unaffected by the treatment modality. CONCLUSION: General anesthesia with propofol and desflurane facilitates shorter induction and recovery times without adversely affecting patient comfort. Therefore, this technique may be preferable to spinal anesthesia for elderly patients undergoing short transurethral surgical procedures. PMID- 9873963 TI - Hemodynamic changes during a new procedure for regional chemotherapy involving occlusion of the thoracic aorta and inferior vena cava. AB - STUDY OBJECTIVE: To describe the hemodynamic consequences of a regional chemotherapy procedure involving occlusion of the thoracic aorta and inferior vena cava (IVC) by intraluminal balloons. DESIGN: Prospective study. SETTING: Operating rooms of an academic hospital. PATIENTS: 10 patients with inoperable intraabdominal malignancy. INTERVENTIONS: After the induction of general anesthesia and the insertion of a pulmonary artery catheter the patients underwent the regional chemotherapy procedure. MEASUREMENTS AND MAIN RESULTS: Occlusion of the thoracic aorta induced an increase in blood pressure (BP) and systemic vascular resistance (SVR) (41% +/- 8% and 80% +/- 15% from baseline, respectively), and a 30% +/- 7% decrease in cardiac output (CO). After aortic balloon deflation at the end of the procedure, we observed a decrease in BP to baseline values, decrease in SVR (to 62% +/- 12% below baseline), and increase in CO (to 80% +/- 15% above baseline). Those changes resemble those described during vascular surgery. Isolated occlusion of the IVC before aortic occlusion caused hemodynamic deterioration in only three of 10 patients, suggesting incomplete obstruction or collateral blood flow in others. Occluding the IVC while the aorta was occluded, caused minimal hemodynamic changes. CONCLUSIONS: Independent inflation of the IVC balloon should not be performed routinely because of possible unpredicted hemodynamic instability. Inferior vena cava occlusion should always be performed after complete aortic occlusion, because it is then that it produces negligible hemodynamic consequences. It is possible that a better assessment of IVC occlusion after balloon inflation needs to be done by contrast injection to prevent a possible leak of chemotherapeutic drugs. PMID- 9873964 TI - Electromyographic assessment of ulnar nerve motor block induced by lidocaine. AB - STUDY OBJECTIVE: To determine the differences in the onset time and duration of motor block produced by lidocaine 1% and lidocaine 2% via a quantitative and objective method, the measurement of compound muscle action potentials (CMAPs). STUDY DESIGN: Prospective study. SETTING: Main operating rooms of a university hospital. PATIENTS: 20 consecutive patients undergoing surgery not requiring intraoperative muscle relaxation. INTERVENTIONS: General anesthesia with unilateral ulnar nerve block was administered. In patients' nondominant (experimental) arms, an insulated block needle was placed adjacent to the ulnar nerve at the wrist while continuous nerve stimulation was delivered to ensure its proper placement. Through this needle, lidocaine 1% or lidocaine 2% was injected. The dominant (control) arm received no injection. MEASUREMENTS AND MAIN RESULTS: Monitoring of ulnar nerve-evoked CMAPs was performed simultaneously on both arms. Ulnar nerve function was assessed at baseline and then at 10-second intervals by automatically measuring the amplitude of the evoked CMAPs on a two-channel electromyogram. The mean (+/- SEM) baseline CMAP amplitude prior to injection of lidocaine 1% was 3.10 +/- 0.87 mV and 3.06 +/- 0.89 mV for the experimental and control ulnar nerves, respectively (p = NS); for lidocaine 2%, baseline CMAP amplitude was 3.58 +/- 1.39 mV and 3.70 +/- 1.46 mV, respectively (p = NS). Over the course of the study, the control CMAP amplitude varied by < 12%. At the experimental ulnar nerve, 90% CMAP decrease after injection of lidocaine 1% and lidocaine 2% occurred 7.5 +/- 2 minutes and 5 +/- 1.5 minutes, respectively (p = NS), whereas maximal blockade was achieved after 15 +/- 3 minutes and 11 +/- 5 minutes, respectively (p = NS). Recovery of CMAP to 90% of baseline occurred 184 +/- 31 minutes after injection of lidocaine 1% and 248 +/- 30 minutes following lidocaine 2% (p = NS). CONCLUSION: The present study describes a technique that can be used in vivo to objectively measure the speed of onset and duration of local anesthetic-induced motor blockade. Although statistically not different, lidocaine 2% demonstrated a faster onset and longer duration of ulnar nerve motor block than lidocaine 1%. PMID- 9873965 TI - Lack of effect of CO2 on cerebral arterial diameter in man. AB - STUDY OBJECTIVE: To establish the quantitative effects on the diameter of cerebral arteries following controlled changes in arterial carbon dioxide tension (PaCO2). DESIGN: Nonrandomized interventional study. SETTING: Angiography suite of a tertiary referral hospital. PATIENTS: 12 anesthetized patients suffering from a cerebral arteriovenous malformation undergoing endovascular treatment. INTERVENTION: Induced hypocapnia by hyperventilation and induced graded hypercapnia by the administration of carbon dioxide to the anesthetized patient's breathing circuit. MEASUREMENTS AND MAIN RESULTS: A digital angiography computer was used to make computerized measurements and calculations of the diameter of deep and small cortical arteries outside the vascular territory of cerebral arteriovenous malformations following controlled and standardized changes in PaCO2. Cardiovascular parameters were simultaneously measured and cardiac output (CO) calculated. No statistically significant changes in the diameter of cerebral arteries down to a size of 0.57 mm, which was the smallest artery studied, could be observed following changes in PaCO2 in the range between 28 +/- 4 mmHg and 74 +/- 4 mmHg. However, there was a 64% change in cardiac index following the above change in PaCO2. CONCLUSION: Deep cortical cerebral arteries down to a diameter of 0.57 mm seem to act merely as conductance vessels. The observed dramatic increase in CO following an increase in PaCO2 may offer an explanation for the changes in cerebral blood flow and cerebral flow velocity recorded by others and usually attributed to cerebral vasodilatation, which we were unable to demonstrate in this study. PMID- 9873966 TI - Physiological dead space/tidal volume ratio during face mask, laryngeal mask, and cuffed oropharyngeal airway spontaneous ventilation. AB - OBJECTIVE: To compare the physiological dead space/tidal volume ratio and arterial to end-tidal carbon dioxide tension (ETCO2) difference during spontaneous ventilation through a face mask, a laryngeal mask (LMA), or a cuffed oropharyngeal airway. DESIGN: Prospective, randomized, cross-over study. SETTING: Inpatient anesthesia at a university department of orthopedic surgery. PATIENTS: 20 ASA physical status I and II patients, without respiratory disease, who underwent ankle and foot surgery. INTERVENTIONS: After a peripheral nerve block was performed, propofol anesthesia was induced and then maintained with a continuous intravenous (i.v.) infusion (4 to 6 mg/kg/h). A face mask, a cuffed oropharyngeal airway, or an LMA were placed in each patient in a random sequence. After 15 minutes of spontaneous breathing through each of the airways, ventilatory variables, as well as arterial, end-tidal, and mixed expired CO2 partial pressure, were measured, and physiological dead space/tidal volume ratio was calculated. MEASUREMENTS AND MAIN RESULTS: Expired minute volume and respiratory rate (RR) were lower with LMA (5.6 +/- 1.2 L/min and 18 +/- 3 breaths/min) and the cuffed oropharyngeal airway (5.7 +/- 1 L/min and 18 +/- 3 breaths/min) than the face mask (7.1 +/- 0.9 L/min and 21 +/- 3 breaths/min) (p = 0.0002 and p = 0.013, respectively). Physiological dead space/tidal volume ratio and arterial to end tidal CO2 tension difference were similar with the cuffed oropharyngeal airway (3 +/- 0.4 mmHg and 4.4 +/- 1.4 mmHg) and LMA (3 +/- 0.6 mmHg and 3.7 +/- 1 mmHg) and lower than with the face mask (4 +/- 0.5 mmHg and 6.7 +/- 2 mmHg) (p = 0.0001 and p = 0.001, respectively). CONCLUSION: Because of the increased dead space/tidal volume ratio, breathing through a face mask required higher RR and expired minute volume than either the cuffed oropharyngeal airway or LMA, which, in contrast, showed similar effects on the quality of ventilation in spontaneously breathing anesthetized patients. PMID- 9873967 TI - Heart rate variability and the prone position under general versus spinal anesthesia. AB - STUDY OBJECTIVE: To evaluate heart rate (HR) variability in the prone position with power spectral heart rate (PSHR) analysis during spinal and general anesthesia. DESIGN: Prospective, clinical evaluation of HR variability in the prone position. SETTING: Tertiary care teaching hospital. PATIENTS: 20 healthy, ASA physical status I and II patients scheduled for elective lumbar spine surgery in the prone position. INTERVENTIONS: Anesthetic technique was either a standard general anesthetic or spinal anesthetic, based on the preference of the patient. Power spectral heart rate, HR, and blood pressure (BP) readings were determined prior to anesthetic intervention and as soon as a stable PSHR reading was available in the prone position. MEASUREMENTS AND MAIN RESULTS: Heart rate and BP were recorded at baseline prior to anesthesia and at the time of stable PSHR data in the prone position. Power spectral heart rate data included low-frequency activity (LFa), high-frequency activity (HFa), and the ratio (LFa/HFa). Spinal anesthesia level was recorded by thoracic dermatome at complete onset. Data were collected from 20 patients; 12 patients chose spinal anesthesia and 8 chose general anesthesia. The prone position resulted in significant increase in HR in the spinal group and significant decrease in BP in the general anesthesia group. Low-frequency activity and LFa/HFa ratio were unchanged in the spinal anesthesia group and were significantly decreased in the general anesthesia group. Spinal level was T8.7. CONCLUSIONS: The association of less change in LFa activity and preservation of BP on assumption of the prone position in patients during low spinal anesthesia suggests better preservation of autonomic nervous system compensatory mechanisms during low spinal anesthesia than with general anesthesia. PMID- 9873968 TI - The effects of preanesthetic oral clonidine on total requirement of propofol for general anesthesia. AB - STUDY OBJECTIVE: To investigate the effects of preanesthetic oral clonidine on total propofol requirement for uniform minor surgery (breast conservative surgery: breast cancer removal with axillary lymph node dissection), and to compare the action of clonidine with that of preanesthetic oral diazepam, a commonly used benzodiazepine. DESIGN: Randomized double-blinded study. SETTING: Operating room ASA physical status I and II room and recovery room of the cancer center. PATIENTS: 80 breast cancer patients scheduled for surgery. INTERVENTIONS: Patients were randomized to one of four treatment groups (placebo, clonidine 75 micrograms, or 150 micrograms of clonidine, or 10 mg of diazepam were orally administered 60 min before induction of anesthesia); n = 20 per group. After evaluating the sedation and anxiety levels of patients using a visual analog scale, anesthesia was induced with propofol (1.5 mg/kg), and maintained with oxygen (O2): nitrous oxide (N2O) (30:70) with a continuous infusion of propofol. The propofol infusion was started at 10 mg/kg/h for 10 minutes, then decreased to 8 mg/kg/h, and 6 mg/kg/h thereafter, and the rate of infusion was adjusted to obtain adequate anesthesia (maintaining hemodynamic parameters within 20% of that prior to premedication). Fentanyl 0.2 mg (each 0.1 mg was given for intubation and axillary lymph node dissection, respectively) was administered. MEASUREMENTS AND MAIN RESULTS: Preanesthetic oral clonidine (150 micrograms) and diazepam (10 mg) induced anxiolysis without sedation. The total requirement (the mean infusion rates) of propofol in placebo, clonidine 75 micrograms, clonidine 150 micrograms, and 10 mg of diazepam groups were 841 +/- 70 (9.0 +/- 0.3), 720 +/- 63 (7.1 +/- 0.4), 491 +/- 39 (5.6 +/- 0.2), and 829 +/- 77 mg (7.9 +/- 0.4 mg/kg/h), respectively. The cost of propofol in these groups was $51.0 +/- 3.8, $45.5 +/- 3.2, $33.5 +/- 2.3, and $50.5 +/- 4.4, respectively. CONCLUSIONS: Preanesthetic oral clonidine (150 micrograms) but not diazepam (10 mg) reduced the total requirement of propofol while stabilizing hemodynamic parameters. In addition, 150 micrograms of oral clonidine attenuates the hemodynamic responses associated with tracheal intubation. PMID- 9873969 TI - Spinal anesthesia for elective lumbar spine surgery. AB - STUDY OBJECTIVE: To evaluate a large series of elective lumbar spine surgical procedures by a single surgeon whose patients were all offered spinal anesthesia. DESIGN: Retrospective chart review. SETTING: Tertiary-care teaching hospital. MEASUREMENTS AND MAIN RESULTS: The records of all elective lumbar spine procedures between 1984 and 1995 performed by one surgeon (GRB) were obtained, and 803 were identified. Of those 803 patients, 611 accepted spinal anesthesia. Data collected included patient demographics, details of the spinal and general anesthesia, perioperative complications, and impact of the spinal anesthetic options on the outcome of spinal anesthesia. General and spinal anesthesia patients were comparable for age, gender, height, and ASA physical status. Patients who received spinal anesthesia were significantly heavier than the general anesthesia patients. Among perioperative complications, nausea and deep venous thrombosis occurred significantly more often in the general than spinal anesthesia patients. Mild hypotension and decreased heart rate (HR) were the most common hemodynamic changes with spinal anesthesia, whereas hypertension and increased HR were the result of general anesthesia. Among spinal anesthetic drugs, plain bupivacaine was associated with the lowest incidence of supplemental local anesthetic use intraoperatively compared to hyperbaric bupivacaine or hyperbaric tetracaine. CONCLUSION: Spinal anesthesia is an effective alternative to general anesthesia for lumbar spine surgery and has a reduced rate of minor complications. PMID- 9873970 TI - Determination of the analgesic dose-response relationship for epidural fentanyl and sufentanil with bupivacaine 0.125% in laboring patients. AB - STUDY OBJECTIVE: To establish the analgesic effective doses as defined as a visual analog pain scale (VAS) of at least 10 for 95% of parturients (ED95) receiving either epidural fentanyl or sufentanil with bupivacaine 0.125% for labor analgesia. DESIGN: Double-blind, randomized controlled study. SETTING: Two tertiary-care teaching hospitals. PATIENTS: 100 female patients, at full-term pregnancy, in active early labor (< 5 cm cervical dilation) and requesting obstetric anesthesia services for labor analgesia. INTERVENTIONS: Patients were randomized and equally distributed to receive one of ten epidural dosing regimens of bupivacaine 0.125% alone or with either fentanyl 25, 50, 75, or 100 micrograms or sufentanil 5, 10, 15, 20, or 25 micrograms in a 10-ml bolus after a 3-ml test dose of bupivacaine 0.25%. MEASUREMENTS AND MAIN RESULTS: VAS scores were obtained from each parturient using a 10-cm plastic VAS slide rule at times 0, 1, 5, 10, 15, 20, 25, and 30 minutes, and then again when the patient requested additional analgesia. Analgesic duration and demographic and obstetric data also were obtained. Using a log-probit dose-response analysis, analgesic success as defined as a VAS of at least 10 with each opioid dose was plotted and an ED95 value of 8 micrograms and 50 micrograms was established for sufentanil and fentanyl, respectively, in bupivacaine 0.125%. No statistical difference was detected for analgesic duration or incidence of side effects between analgesic groups. CONCLUSIONS: Epidural analgesia with fentanyl and sufentanil in bupivacaine 0.125% behaves in a dose-response fashion allowing for the determination of equipotent dose of each. PMID- 9873971 TI - Massive contralateral pneumonia following thoracotomy for lung resection. AB - Separation of the lungs with a double-lumen endobronchial tube facilitates the surgical approach and protects the bronchial system from contamination by pus or blood from the operated lung. We report a case of a 49-year-old man who suffered a gram-negative pneumonia requiring mechanical ventilatory support and prolonged hospitalization after pulmonary resection. Contamination of the ventilated dependent lung was documented intraoperatively. Current information on postoperative infectious complications and preventive measures are discussed. PMID- 9873972 TI - Bullard laryngoscope: preliminary experience with the new multifunctional stylet. PMID- 9873973 TI - 'Wrongful life' claims. PMID- 9873974 TI - Paid organ donation--the grey basket concept. PMID- 9873975 TI - 'Wrongful life' lawsuits for faulty genetic counselling: should the impaired newborn be entitled to sue? AB - A "wrongful life" suit is based on the purported tortious liability of a genetic counsellor towards an infant with hereditary defects, with the latter asserting that he or she would not have been born at all if not for the counsellor's negligence. This negligence allegedly lies in the failure on the part of the defendant adequately to advice the parents or to conduct properly the relevant testing and thereby prevent the child's conception or birth (where unimpaired life was not possible). This paper will offer support for the thesis that it would be both feasible and desirable to endorse "wrongful life" compensation actions. The genetic counsellor owed a duty of due professional care to the impaired newborn who now claims that but for the counsellor's negligence, he or she would not have been born at all. The plaintiff's defective life (where healthy life was never an option) constitutes a compensable injury. A sufficient causal link may exist between the plaintiff's injury and the defendant's breach of duty of due professional care and an appropriate measure of damages can be allocated to the disabled newborn. Sanctioning a "wrongful life" cause of action does not necessarily entail abandoning valuable constraints with regard to abortion and euthanasia. Nor does it inevitably lead to an uncontrolled slide down a "slippery slope". PMID- 9873976 TI - Transfusion-free treatment of Jehovah's Witnesses: respecting the autonomous patient's motives. AB - What makes Jehovah's Witnesses tick? What motivates practitioners of medicine? How is benevolent human behaviour to be interpreted? The explanation that fear of censure, mind-control techniques or enlightened self-interest are the real motivators of human conduct is questioned. Those who believe that man was created in "God's image", hold that humanity has the potential to rise above selfishly driven attitudes and actions, and reflect the qualities of love, kindness and justice that separate us from the beasts. A comparison of general medical ethics and disciplines, and those of the Jehovah's Witness community, is made in this context. The easy charge that frequent deaths result from refusal of blood transfusions is examined. The central source of antipathy towards Jehovah's Witnesses, namely the alleged imposition of extreme and even harmful refusal of blood therapy on our children is addressed. Of course, "...few dilemmas are likely to be resolved wisely or satisfactorily by a blinkered adherence to abstract principles alone. Solutions to most cases will be dictated by a combination of factors. The support of medical ethics by Jehovah's Witnesses, and their willingness to share in reasoned and ethical debate, while at the same time holding firm to their religious and conscientious principles are emphasised. PMID- 9873977 TI - Two worlds apart: religion and ethics. AB - In a recent article entitled, Requests "for inappropriate" treatment based on religious beliefs, Orr and Genesen claim that futile treatment should be provided to patients who request it if their request is based on a religious belief. I claim that this implies that we should also accede to requests for harmful or cost-ineffective treatments based on religious beliefs. This special treatment of religious requests is an example of special pleading on the part of theists and morally objectionable discrimination against atheists. It also provides an excellent illustration of how different the practices of religion and ethics are. PMID- 9873978 TI - Medicine, ethics and religion: rational or irrational? AB - Savulescu maintains that our paper, which encourages clinicians to honour requests for "inappropriate treatment" is prejudicial to his atheistic beliefs, and therefore wrong. In this paper we clarify and expand on our ideas, and respond to his assertion that medicine, ethics and atheism are objective, rational and true, while religion is irrational and false. PMID- 9873979 TI - The commodification of human reproductive materials. AB - This essay develops a framework for thinking about the moral basis for the commodification of human reproductive materials. It argues that selling and buying gametes and genes is morally acceptable although there should not be a market for zygotes, embryos, or genomes. Also a market in gametes and genes should be regulated in order to address concerns about the adverse social consequences of commodification. PMID- 9873980 TI - Tell me what's wrong with me: a discourse analysis approach to the concept of patient autonomy. AB - BACKGROUND: Patient autonomy has gradually replaced physician paternalism as an ethical ideal. However, in a medical context, the principle of individual autonomy has different meanings. More knowledge is needed about what is and should be an appropriate understanding of the concept of patient autonomy in clinical practice. AIM: To challenge the traditional concept of patient autonomy by applying a discourse analysis to the issue. METHOD: A qualitative case study approach with material from one consultation. The discourse is interpreted according to pragmatic and text-linguistic principles and provides the basis of a theoretical discussion of different concepts of patient autonomy. RESULTS: The consultation transcript illustrates how the patient's wishes can be respected in real life. The patient, her husband and the doctor are all involved in the discourse dynamics, governed by the subject matter, namely her mental illness. CONCLUSION: We suggest a dynamic and dialogue-based conception of autonomy as adequate for clinical purposes. These perspectives, based on mutual understanding, take communication between patient and doctor as their starting point. According to this approach, autonomy requires a genuine dialogue, an interpersonal mode of being which we choose to call "authentic interaction". PMID- 9873981 TI - Can unequal be more fair? Ethics, subject allocation, and randomised clinical trials. AB - Randomised clinical trials provide the most valid means of establishing the efficacy of clinical therapeutics. Ethical standards dictate that patients and clinicians should not consent to randomisation unless there is uncertainty about whether any of the treatment options is superior to the others ("equipoise"). However, true equipoise is rarely present; most randomised trials, therefore, present challenging ethical dilemmas. Minimising the tension between science and ethics is an obligation of investigators and clinicians. This article briefly reviews several techniques for addressing this issue and suggests that unbalanced randomisation, a technique rarely employed in current clinical trial practice, may be useful for enhancing the ethical design of human experimentation. PMID- 9873982 TI - Does pregnancy affect medical ethical decision making? AB - OBJECTIVE: We studied and compared the attitudes of pregnant women v new mothers in an attempt to confirm changing patterns of maternal response towards medical ethical decision making in critically ill or malformed neonates. DESIGN: Data were obtained by questionnaires divided into three sections: 1. sociodemographic; 2. Theoretical principles which might be utilised in the decision-making process; 3. Hypothetical case scenarios, each followed by possible treatment options. RESULTS: Pregnant women (n = 545) consistently requested less aggressive medical intervention for the hypothetical cases than did new mothers (n = 250) [Trisomy 18: 57% v 42%; p = 0.0004; Asphyxia: 75% v 63%; p = 0.0017; Down's syndrome 81% v 62%; p = 0.0001; LBW 85% v 75%; p = 0.004]. Significant differences were also observed in the responses to the theoretical principles, with pregnant women attributing less importance to preserving life at all cost, while being more concerned with physical and emotional pain and suffering, with financial cost, and with the infant's potential for future productivity. PMID- 9873983 TI - On autonomy and identity. PMID- 9873984 TI - The ethics of xenotransplantation. PMID- 9873985 TI - Bibliography section. PMID- 9873987 TI - Characterization of Schistosoma mansoni specific human T-T hybrids. AB - In this study, we generated antigen-specific T-T hybrids by the fusion of an established T lymphoma, CEM, with T-cell lines derived from Schistosoma mansoni antigen-stimulated peripheral blood mononuclear cells (PBMC) of schistosomiasis patients. These PBMC were capable of in vitro proliferation and granuloma formation upon proper stimulation with antigens derived from different phases of S. mansoni development. Culture supernatants collected from T-T hybrids were responsible for the stimulation or inhibition of antigen-specific responses of other PBMC, on in vitro proliferation and granuloma reaction assays. Clones derived from a T-T hybrid were characterized as CD4+ CD8- HLA-DR- and expressed the cellular markers CD3 and CD25, especially after antigenic stimulation. Their supernatants, as that from the T-T hybrid line, were capable of modulation of in vitro cellular responses, and IL-2, IL-10, and TNF-alpha were detected upon cellular stimulation with antigen or phytohemagglutinin-A (PHA). PMID- 9873986 TI - Characteristic immunolocalization of Ku protein as nuclear matrix. AB - Two hybridoma clones, NMB1 and NML90, were established using nuclear matrix proteins from normal human thymi or malignant lymphoma as immunogens. They reacted with human Ku70 and Ku80, respectively, by immunoblotting. When HeLa cell nuclear proteins were fractionated and applied to immunoblotting, both Ku70 and Ku80 were detected in the nuclear matrix as well as the soluble nuclear protein fractions. By confocal scanning microscopy, the immunoreactivity of Ku70 and Ku80 was localized to distinct nucleoplasmic fibrillar network and fine granules in the interphase cell nuclei. When HeLa cells were fractionated in situ using DNase I and buffers containing 0.25 M (NH4)2SO4 and 2 M NaCl, the nucleoplasmic reticular structure was largely preserved, but granules disappeared. The nucleoplasmic distribution of Ku in the tissue and in cultured cells was distinct from each other. In the adult tissue, it consisted mostly of either distinct curvilinear lines along the nuclear periphery or of tangled, beaded lines throughout the nuclei. When xenotransplants of HeLa cell in Scid mice were examined, the "tissue type" immunolocalization pattern was reproduced consistently. In most fetal tissues, "tissue type" and "cell type" patterns were admixed. Monoclonal antibodies described here are useful tools for studying the structure and function of the nuclear matrix. PMID- 9873988 TI - Autoantibodies in the serum of patients with gastric cancer: their prognostic importance. AB - To evaluate the presence in serum and the clinical relevance of several antinuclear autoantibodies, we investigated 31 patients with initially diagnosed gastric cancer and 40 age-matched healthy controls. Autoantibodies against ssDNA, dsDNA, cardiolipin, actin, myosin, tropomyosin, GM1, GD1b and GT3 gangliosides, were detected with an enzyme-linked immunoassay (ELISA). Anti-ssDNA, anti-actin, anti-GM1 and anti-GD1b antibodies were detected in the serum of 11 (p = 0.001), 8 (p = 0.02), 11 (p = 0.001), and 9 (p = 0.008) patients with gastric cancer, respectively. There was no significant difference between patients with cancer and the control group, as far as the other autoantibodies were concerned. Most of the patients (90%) had autoantibodies against at least one of the antigens examined. Patients with anti-ssDNA, anti-actin, anti-GM1 and anti-GD1b antibodies were less likely to survive than the patients being negative to the above autoantibodies: the figures are 1 of 11 (9%) compared with 4 of 20 (20%); 1 of 8 (13%) compared with 5 of 23 (22%); 1 of 11 (9%) compared with 4 of 20 (20%); and 1 of 9 (11%) compared with 4 of 22 (18%), respectively. Our findings suggest that 4 of the 9 autoantibodies that we assayed are significantly more likely to be found in serum of patients with gastric cancer, indicating that the immune system has a role in the process of the malignant disease. If our results are confirmed by forthcoming studies, some of the immunological variables that we examined could be used as markers of prognostic value in patients with gastric cancer. PMID- 9873989 TI - Characterization of a novel monoclonal antibody raised against human hepatocellular carcinoma. AB - A monoclonal antibody (MAb) targeting human hepatocellular carcinoma (HCC) has been produced by immunizing BALC/c mice with HCC SMMC-7721 cells. Solid phase enzyme-linked immunoabsorbent assay (ELISA) analysis showed that this mAb (mAb 95) was reactive with human HCC cell lines and tumor biopsies, but not tumors of other origin or normal tissues tested. Immuno-gold labeling and flow cytometry indicated that mAb 95 bound specifically to HCC cell membrane protein. Western blot analysis showed mAb 95 reacts only with HCC cell lysates, but not with 10 different human normal tissue lysates. The approximate molecular size of the reactive proteins were 40,000 and 60,000 daltons. The specificity of mAb 95 indicates it may be a potential candidate for a targeting agent in HCC immunotherapy or immunodetection. PMID- 9873991 TI - A novel monoclonal antibody against rodent common Thy-1.1 epitope. AB - Here we report a new anti-rat Thy-1 monoclonal antibody (MAb) 12C5 that recognized a novel epitope on a GPI-anchored Thy-1 antigen. In rat thymocytes, MAb 12C5 reacted with 25 kDa Thy-1 antigen purified with MAb OX7. In rats, MAb 12C5 was expressed on 94% of thymocytes, 46% of spleen cells, 31% of mesenteric lymphnode cells, 76% of bone marrow cells, and 24% of splenic T cells. Furthermore, MAb 12C5 reacted with the hematopoietic cells in Mongolian gerbils, hamsters, and AKR mice (Thy-1.1) as in rats. In AKR mice, almost all bone marrow cells reacted with MAb 12C5. Thus, MAb 12C5 recognized a novel epitope of Thy-1 that was widespread in hematopoietic cells of rats and mice, as well as other rodents. PMID- 9873990 TI - Novel monoclonal antibodies to putative selectin carbohydrate ligands that inhibit selectin binding to myeloid cells. AB - Four newly developed monoclonal antibodies (MAbs) are characterized using flowcytometry, enzyme-linked immunoadsorbent assay (ELISA), immunoprecipitation and Western blots, carbohydrate epitope mapping, glycosidase cleavage, and competition binding assays. Their effects on selectin binding to myeloid cells was tested. These MAbs react only with myeloid cells. MAbs CI-1, BU60, and HIM95 recognize epitopes expressed by CD11/CD18 (beta2) integrins, while HI247 and CSLEX1 do not. The epitopes require Lewis x [Galbeta1-4 (Fucalpha1-3)GlcNAc] based on reactivity with oligosaccharide-polyacrylamide-biotin or oligosaccharide BSA conjugates. MAb HI247 recognizes a related structure, sialyl-Lewis x, NeuAcalpha2-3GaLbeta1-4(Fucalpha1-3)GlcNAc. The three MAbs against Lewis x show some minor differences in their reactivity such as recognizing their antigens on CD11/CD18 integrins after endo-beta-galactosidase treatment and recognizing free Lewis x. The hydroxyl group on C-3 of the terminal galactose is important for recognition by MAb CI-1, BU60, and HIM95 as its substitution with sulfo group of sialic acid abolishes the binding of these MAbs. The C-3 sialic acid is crucial for the binding of MAb HI247. Its replacement by sulphate or its cleavage by sialidase eliminates recognition by this MAb. MAbs HI247 and CSLEX-1 did not react in ELISA with immobilized CD11/CD18, suggesting that the majority of sialyl Lewis x on CD11/CD18 molecules may have sialic acid 6-linked rather than 3-linked to galactose. Unexpectedly, MAb BU60 inhibited binding of P-selectin mu chain chimera to HL-60 or U937 cells, while CI-1, HIM95 and three other defined anti Lewis x MAbs (6C7, M6-1 and LeuM1) did not. MAb HI247 inhibited binding of both E and P-selectin chimeras to these cell lines more effectively than several characterized MAbs (CSLEX-1, FH6, HECA-452) to sialyl Lewis x and related oligosaccharides. Certain combinations of these anticarbohydrate MAbs had additive inhibitory effects on selectin binding, suggesting a potential application of these new MAbs in cell adhesion/migration and tumor metastasis studies. PMID- 9873992 TI - Characterization of an immunosuppressive anti-CD40 ligand monoclonal antibody. AB - The interaction between CD40 ligand (CD40L) and its counter-receptor CD40 is critically important in T- and B-cell costimulation and generation of the humoral immune response. But several questions still remain unsolved, particularly in the human in vivo system. To clarify the precise function of CD40L and develop an immunosuppressive agent, we have generated a murine monoclonal antibody (MAb), 2B2 specific for human CD40L. The specificity of this MAb for human CD40L was verified by enzyme-linked immunoadsorbent assay (ELISA) and flow cytometry. MAb 2B2 immunoprecipitated proteins of molecular weight 35 and 28 kD on human peripheral blood lymphocytes (PBLs) stimulated with phorbol 12-myristate-13 acetate (PMA) plus ionomycin. Then we have studied the biological effect of MAb 2B2 in severe combined immunodeficiency (SCID) mice reconstituted with human PBLs. The data showed that this MAb strongly suppressed human IgG production of human B cells transplanted in SCID mice, indicating that this MAb 2B2 could be used to regulate unwanted immune responses associated with autoimmune disease. Then we analyzed the sequence of MAb 2B2. The 2B2 heavy chain variable region (VH) and light chain variable region (VL) genes were cloned using PCR. The cloned VH gene coded for 123 amino acid residues and belonged to the subgroup III(D). The VL gene coded for 126 amino acid and belonged to the subgroup V. Collectively, these results will be used to develop an immunosuppressive chimeric or humanized anti-CD40L antibody. PMID- 9873993 TI - Evaluation of the affinity measurement of anti-HIV-1 p17 monoclonal antibody by BIAcore. AB - The methods of measuring the affinity constants of anti-HIV-1 p17 monoclonal antibodies (MAbs) using the double antibody methods in the liquid phase and the biomolecular interaction analysis by BIAcore system (Pharmacia Biosensor AB, Uppsala, Sweden) were compared. MAbs, HyHIV1-6, recognizing residues 12-29 (P12 29) of p17 and the naive protein, p17, were used. The kinetic association constants (KAs) obtained using the double antibody method were 2.40 x 10(7) - 1.40 x 10(8)M(-1) for P12-29, and 4.80 x 106 - 1.80 x 10(7) M(-1) for p17. In the BIAcore system where P12-29 or p17 was used as immobilized antigens onto the sensorchip, the KAs were 1.57 x 10(9) - 4.81 x 10(9) M(-1) for P12-29, and 1.52 x 10(9) - 1.21 x 10(10) M(-1) for p17. On the other hand, when MAbs were immobilized onto the sensorchip and P12-29 or rp17 was used as analyte, the KAs for P12-29 and p17 were in the region 3 x10(8) - 3 x 10(9), 1 x 10(8) - 3 x 10(9) M(-1), respectively. These data show that the KAs were higher than those obtained using the double antibody method, however, no significant difference could be observed. Moreover, the KAs obtained for p17 using MAbs as ligand were similar for BIAcore and the double antibody method except for HyHIV2. Therefore, the BIAcore system can be used for the affinity measurement instead of the double antibody method. PMID- 9873994 TI - Long-term production of human monoclonal antibodies by five human-mouse heterohybridomas in a serum-free medium. AB - Four heterohybridomas (28K29, 27D57, ZLG40, and 29D38) secreting human monoclonal antibodies (MAbs) reactive with lung cancer cells and one heterohybridoma (28B49) secreting an isotype-matched MAb (IgM, lambda) were adapted and tested for long term MAb production in serum-free medium. Three typical serum-free media were first evaluated for this purpose, based on the MAb productivity, cost, and ease of MAb purification with each, and Hybridoma-SFM, containing only insulin and transferrin as protein components, was selected as the most appropriate. In this medium, all five heterohybridomas continued to proliferate and secrete MAb throughout test periods of 144 to 200 days. Achievement of this long-term, stable production may be attributed at least in part to the effects of extensive prior subcloning and to the predominance of lambda-type light chains in the IgM antibodies produced by these heterohybridomas. The results of this study provide the basis for development of a process to produce large quantities of human MAbs, which may be useful for therapeutic or in vivo diagnostic purposes. PMID- 9873995 TI - Anti-TRAP 14G6 is effective for immunochemistry. PMID- 9873996 TI - Stress-induced morphohistochemical and functional changes in rat adrenal cortex, testis and major salivary glands. AB - The effect of repeated stress (1 h of daily immobilization for seven consecutive days) on the adrenal cortex of young adult male albino rats was evaluated by morphohistochemical methods and plasma assays; at the same time, testes and major salivary glands, as steroid-producing and -depending organs, respectively, were examined. Morphological and histochemical changes were found in the adrenal cortex, testis and submaxillary gland, though varying in degree and extent depending on the gland examined. Corticosterone and progesterone plasma levels increased, in agreement with the lipid depletion observed in the zona fasciculata, while testosterone and androstenedione decreased, as confirmed by the less marked enzymatic activity in the Leydig cells. The study thus proves that repeated stress, even of temporary duration, is able to influence directly or indirectly the morphofunctional state of the three examined glands, suggesting a functional linkage. PMID- 9873997 TI - Microwave staining of intestinal whole-mount preparations with Cuprolinic blue. AB - Encouraged by the knowledge that microwaves have a beneficial effect on immunohistochemical reactions, the present study aimed to find out whether microwaves could improve the Cuprolinic Blue staining of enteric neurons as well as the actual method that has been developed for gastrointestinal whole-mount preparations. In addition to incorporating a microwave application in the method described by Holst and Powley (1995), some other modifications were made: two incubations before incubation in the staining solution and free-floating incubations. In the whole-mount preparations, most, if not all, enteric neurons were stained by Cuprolinic Blue. These neurons appeared as blue-green cells with non-reacting nuclei and neuronal processes. At higher magnification, the cytoplasm was characterized by a fine blue-green granulation, and the nucleolus in the nucleus appeared as a blue iridescent structure. Non-specific staining occurred in fibrocytes and epithelial cells but, because of their location and appearance, they could easily be distinguished from neurons. The modified incubations and the incorporation of a microwave application into the conventional Cuprolinic Blue staining method turn the method into an easy-to-use one that seems to visualize most, if not all, enteric neurons in whole-mount preparations of the pig jejunum. PMID- 9873998 TI - Non-identical distribution pattern of epidermal growth factor and platelet derived growth factor in the mouse uterus during the oestrous cycle and early pregnancy. AB - In the present study, we examined by immunohistochemistry the cell-specific distribution of epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) in the mouse uterus during the oestrous cycle and throughout the first 7 days of pregnancy. Paraffin-embedded tissue samples were immunostained using the avidin-biotin peroxidase technique and then examined by light microscopy. Our results showed that immunostaining for EGF was detected in the stroma but not in the luminal or glandular epithelium. A high concentration of EGF was detected in the stroma around the time of embryo implantation at days 3, 4 and 5 of pregnancy. The implanted embryo at day 7 of gestation showed immunostaining for EGF between the ectoderm and endoderm layers. The cell distribution pattern for PDGF was found to be different from that observed with EGF. Luminal and glandular epithelia displayed PDGF immunostaining throughout the first 7 days of pregnancy, with the highest intensity at days 4 and 5 of gestation. In contrast, no immunostaining was observed in the luminal and glandular epithelia at post oestrus, dioestrus and pro-oestrus stages. However, a weak reaction started to appear at oestrus. The embryo at the blastocyst stage displayed a strong immunoreaction for antibody against PDGF. In addition, the decidual boundary zone surrounding the implanted embryo at days 5, 6 and 7 of gestation also showed an immunostaining for PDGF. The present observations demonstrate clearly the presence of EGF and PDGF in the mouse uterus in high concentrations at the peri implantation period. Thus, our results, together with what is known about the effect of EGF and PDGF in controlling the growth, differentiation and activation of a variety of cell types, suggest a possible role for these growth factors during the preparation of the endometrium for implantation in controlling the proliferation activity of stromal and/or epithelial cells. PMID- 9874000 TI - Steroid 5alpha-reductase type 1 immunolocalized in the rat peripheral nervous system and paraganglia. AB - Steroid 5alpha-reductase is an enzyme that converts a number of steroids with a C 4, 5 double bond and C-3 ketone to 5alpha-reduced metabolites. This enzyme has been suggested to play a role in brain development and myelination in the rat nervous system. In the present study, we examined the cellular and subcellular localization of the enzyme immunocytochemically in the rat peripheral nervous system and paraganglia using a polyclonal antibody against rat 5alpha-reductase type 1. Light and electron microscopical studies localized 5alpha-reductase in the Schwann cells of myelinated and unmyelinated nerve fibres, the satellite cells of the ganglia, the enteric glial cells and the supporting/sustentacular cells of the paraganglia. In the myelinated nerve fibres, immunoreactivity was observed in the outer loops, the nodes of Ranvier and the Schmidt-Lanterman incisures. Subcellularly, the immunoreactivity was localized in the cytoplasm of various glial cells. No immunoreactivity was observed in the myelin membrane, the axon or the neuronal perikaryon. These findings suggest that 5alpha-reductase is widely distributed in glial cells, and that, in addition to myelination, 5alpha reduced steroids play a role in some glial functions in the peripheral nervous system. PMID- 9873999 TI - Epithelial glycoprotein-2 expression is subject to regulatory processes in epithelial-mesenchymal transitions during metastases: an investigation of human cancers transplanted into severe combined immunodeficient mice. AB - The human cell-surface antigen epithelial glycoprotein-2 recognized by the monoclonal antibody MOC-31 is an epithelial tumour-associated glycoprotein expressed in non-squamous carcinomas. MOC-31 immunoreactivity was investigated in human breast, colon, ovarian and lung cancer cell lines, grown either in vitro or in severe combined immunodeficient (SCID) mice as solid tumours and/or metastases. Three of four small-cell lung cancer cell lines (NCI-H69, OH3 and SW2) and three of four ovarian cancer cell lines (SoTu 1, 3 and 4) expressed epithelial glycoprotein-2. In contrast, all three breast (MCF-7, BT20, T47D) and all three colon (HT29, CACO2, SW480) cancer cell lines strongly reacted with monoclonal antibody MOC-31. A notable difference in MOC-31 immunoreactivity was observed in spontaneously formed lung metastases of HT29 colon cancer cells. Whereas larger metastases (> 30 cells) reacted with a similar staining pattern to the primary tumour, smaller metastases did not. These findings indicate that differentiation processes during the epithelial-mesenchymal transition occur in metastases, which lead to a transient loss of epithelial glycoprotein-2 expression during the migratory and early post-migratory period. This loss of antigen expression indicates that the process of metastases formation is a regulatory event, and this transient loss of antigen expression might represent a potential obstacle to antibody-based therapy in the setting of minimal residual disease. PMID- 9874001 TI - Immunohistochemical localization of arginase II and other enzymes of arginine metabolism in rat kidney and liver. AB - Arginine is a precursor for the synthesis of urea, polyamines, creatine phosphate, nitric oxide and proteins. It is synthesized from ornithine by argininosuccinate synthetase and argininosuccinate lyase and is degraded by arginase, which consists of a liver-type (arginase I) and a non-hepatic type (arginase II). Recently, cDNAs for human and rat arginase II have been isolated. In this study, immunocytochemical analysis showed that human arginase II expressed in COS-7 cells was localized in the mitochondria. Arginase II mRNA was abundant in the rat small intestine and kidney. In the kidney, argininosuccinate synthetase and lyase were immunostained in the cortex, intensely in proximal tubules and much less intensely in distal tubules. In contrast, arginase II was stained intensely in the outer stripes of the outer medulla, presumably in the proximal straight tubules, and in a subpopulation of the proximal tubules in the cortex. Immunostaining of serial sections of the kidney showed that argininosuccinate synthetase and arginase II were colocalized in a subpopulation of proximal tubules in the cortex, whereas only the synthetase, but not arginase II, was present in another subpopulation of proximal tubules. In the liver, all the enzymes of the urea cycle, i.e. carbamylphosphate synthetase I, ornithine transcarbamylase, argininosuccinate synthetase and lyase and arginase I, showed similar zonation patterns with staining more intense in periportal hepatocytes than in pericentral hepatocytes, although zonation of ornithine transcarbamylase was much less prominent. The implications of these results are discussed. PMID- 9874002 TI - Immunohistochemical detection of vitellogenin in male brown trout from Swiss rivers. AB - The detection of vitellogenin, a yolk precursor protein, may serve as a biomarker for exposure to environmental oestrogens as its induction by xenobiotic oestrogens in the immature and male fish has been reported repeatedly. In the present work, juvenile brown trout were injected with oestradiol (5 microg g(-1) body weight oestradiol benzoate) in order to assess the induction and organ distribution of vitellogenin by means of immunohistochemistry. In addition, brown trout collected from Swiss rivers were analysed. Vitellogenin was detected in the oestradiol-injected juvenile trout but not in uninjected controls. The presence of vitellogenin was also demonstrated in a male and an immature feral brown trout from one of two locations downstream of three sewage treatment plants. In contrast, no positive staining was found in livers of trout upstream of the respective plants. The results demonstrate the suitability of immunohistochemistry for monitoring feral fish for the presence of vitellogenin production. PMID- 9874003 TI - Chronic pain: a public health problem? PMID- 9874004 TI - AAPM president's message. Total pain care--new concept? American Academy of Pain Medicine. PMID- 9874005 TI - Complex regional pain syndrome I (CRPS I): prospective study and laboratory evaluation. AB - OBJECTIVE: To relate clinical features to autonomic laboratory indices used in the diagnosis of Complex Regional Pain Syndrome type I (CRPS I) (reflex sympathetic dystrophy) to generate improved diagnostic criteria. DESCRIPTION: CRPS I is a chronic pain syndrome, characterized by diffuse limb pain with allodynia and prominent vasomotor and sudomotor dysfunction. METHODS: We conducted a prospective study on 102 patients referred for possible CRPS I. These patients completed a structured questionnaire and underwent neurologic examination, with special attention to the evaluation of clinical features of vasomotor, sudomotor, motor, and sensory, including pain, dysfunction. All patients were tested using a standard autonomic protocol that compared side-to side skin temperature, resting sweat output, and quantitative sudomotor axon reflex test (QSART) measurements. Composite autonomic clinical (CRPS-Sx) and laboratory (CRPS-LAB) scores were defined. The clinical (subjective and objective) and the laboratory data were analyzed using Pearson's correlation analysis and Bonferroni's probability value to assess concordance and their value in correctly diagnosing CRPS I. RESULTS: All cases occurred after limb injury. One-third of cases did not fulfill our criteria of CRPS I. Highly significant correlations (p<.001) were found among certain clusters of symptoms and signs that shared unifying pathophysiologies. CRPS-Sx correlated with CRPS-LAB (p = .035). The indices that correlated most reliably with clinical data and with each other were RSO, QSART, and skin temperature reductions. CONCLUSION: Clinical and autonomic laboratory probability scores correlate in an internally consistent manner. Both CRPS-Sx and CRPS-LAB are sensitive and reliable tools to formulate a correct diagnosis of CRPS I and can be combined to provide an improved set of diagnostic criteria for CRPS I. PMID- 9874006 TI - Stiff upper lip: coping strategies of World War II veterans with phantom limb pain. AB - OBJECTIVE: Study of coping with phantom pain in nonclinical war veteran amputees. DESIGN: Semistructured interview with amputees in their home setting. SETTING: Residential home for war veteran amputees or respondents' own homes. PATIENTS: Amputee veterans of World War II with phantom pain. OUTCOME MEASURES: Pain (McGill Pain Questionnaire) and pain history, coping (daily coping; Stone and Neale, J Pers Soc Psychol 1984;46:892-906), size of social network, and quality of war memories. RESULTS: No differences in pain or coping were associated with place of residence (and prevalence of cues) or social networks; war memories appeared not to be associated with availability of cues, whether media coverage or other amputees. There was some association between the emotional tone of war memories and pain intensity. Veteran amputees were in general accepting of high levels of pain and made little use of medical resources, relating that to past experience of their pain being dismissed. CONCLUSION: Coping with phantom pain in war veteran amputees is predominantly silent acceptance of the pain, with little use of social support however available, and rare recourse to medical help, based on past unhelpful experience. Pain and mood appeared to be unrelated to specific war cues, but higher pain scores were reported by those with unhappier war memories. PMID- 9874007 TI - Personality assessment of patients with complex regional pain syndrome type I. AB - OBJECTIVE: There is controversy regarding the importance of psychological/psychiatric factors in the development of the Complex Regional Pain Syndrome (CRPS). Our objective was to determine whether CRPS type I patients were psychiatrically different from other chronic pain patients, with particular attention to personality pathology. DESIGN: A standardized clinical assessment of all major psychiatric categories, including personality disorders, was performed on 25 CRPS type I patients and a control group of 25 patients with chronic low back pain from disc-related radiculopathy. MEASURES: Both sections of the Structured Clinical Interview for the Diagnostic and Statistical Manual (3rd ed., rev.) and the visual analog scale. RESULTS: Both groups were similar in terms of pain intensity and duration. Statistical analysis showed both groups to have a significant amount of major psychiatric comorbidity, in particular major depressive disorder, and a high incidence of personality disorders. Therefore, intense chronic pain was associated with significant psychiatric comorbidity in both groups and in similar proportions. CONCLUSION: The high incidence of personality pathology in both groups may represent an exaggeration of maladaptive personality traits and coping styles as a result of a chronic, intense, state of pain. PMID- 9874008 TI - Comparison of the effects of perceived self-efficacy on coping with chronic cancer pain and coping with chronic low back pain. AB - OBJECTIVE: The purposes of this study were to explore the differences between chronic cancer pain and chronic low back pain with respect to (a) the use of coping strategies to manage pain and (b) the relationship between self-efficacy for attenuating pain and pain outcomes. DESIGN: Descriptive correlational design. PATIENTS: Eighty-five patients experiencing chronic low back pain (n = 85) and 88 patients with chronic cancer pain (n = 88) were recruited for this study. OUTCOME MEASURES: Modified Coping Strategies Questionnaire, self-efficacy expectancies, and the Brief Pain Inventory. RESULTS AND CONCLUSIONS: The major findings of this study were that (a) patients with chronic cancer pain reported significantly lower pain intensity and pain interference than did patients with chronic low back pain; (b) the most frequently used coping strategies were almost the same between the low back pain group and the cancer pain group; (c) for both chronic cancer pain and chronic low back pain groups, patients' perceived self-efficacy was significantly inversely correlated with pain intensity and pain interference with daily life; and (d) patients' use of coping strategies was positively correlated with pain intensity and pain interference with daily life. These findings were discussed in terms of implications for clinical practice and future research. PMID- 9874009 TI - Self-reported sleep and mood disturbance in chronic pain patients. AB - OBJECTIVE: To evaluate the prevalence of self-reported sleep disturbance and its relationship to mood disturbance in chronic pain patients. DESIGN AND SETTING: Survey of patients referred to a multidisciplinary outpatient pain clinic. PATIENTS: The sample consisted of 105 consecutive patients (59 men and 46 women), with an average age of 41.5 (SD +/-13.4) years. MEASURES: Self-report measures of sleep disturbance and visual analog scales of mood disturbance (anxiety and depression) and pain experience (intensity and unpleasantness). RESULTS: Patients were grouped according to whether they considered themselves "poor" (n = 68) or "good" (n = 37) sleepers. Poor sleepers reported more difficulties initiating and maintaining sleep and greater pain intensity and pain unpleasantness than did good sleepers. The two groups did not differ on measures of depressive or anxious mood. CONCLUSION: The results suggest that sleep disturbance is a prevalent complaint in chronic pain patients, but it is not always associated with an underlying mood disturbance. PMID- 9874010 TI - Long-term home treatment with epidural analgesia does not affect later spinal cord stimulation in patients with otherwise intractable angina pectoris. AB - OBJECTIVE: The only way some patients with intractable angina pectoris can endure the daily pain is by using opiates. Epidural morphine or spinal cord stimulation (SCS) for out-patients is a possibility for this patient group. The aim of this study was to determine whether patients previously treated with epidural injections had more frequent electrode migration, higher stimulation needs, and less effect of SCS treatment. DESIGN: A prospective, nonrandomized study with a 4 year follow-up period. SETTING: Department of Anaesthesia, Pain Section. A referral center in institutional practice. Ambulatory care. PATIENTS: The patient group comprised 53 patients; all had been treated daily with opiates. A total of 26 patients had epidural catheters for a mean time of 1 year before SCS. All patients had intractable angina pectoris. For these patients, further angioplasty or coronary bypass surgery was not technically possible. Even with maximal medication, it was impossible to cope with the patients' angina pectoris, and the only way the patients could endure the daily pain was by using opiates. Therefore, alternative therapies were considered to give these patients palliation. INTERVENTION: SCS with epidural electrodes stimulating paresthesia in the area where angina is perceived. MAIN OUTCOME MEASURE: Difference for the patients previously treated with epidural catheters in stimulation amplitude, frequency of electrode migration, and effect of SCS. RESULTS: Stimulation demand (p = 0.09), frequency of electrode migrations (p = 0.46), and pain-reducing effect (p = 0.16) were not different for the group of patients previously treated with epidural catheters for longer periods (1-36 months). CONCLUSIONS: SCS in patients previously treated with epidural catheters has an effect equal to that in other patients. PMID- 9874011 TI - Percutaneous electrical nerve stimulation (PENS): a complementary therapy for the management of pain secondary to bony metastasis. AB - OBJECTIVE: To evaluate the use of a novel nonpharmacologic analgesic therapy known as percutaneous electrical nerve stimulation (PENS) in the management of opioid-resistant cancer pain. DESIGN: PENS therapy was administered to three cancer patients on three or more occasion using acupuncturelike needle probes that were stimulated for 30 minutes at frequencies of 4-100 Hz. RESULTS: Two of the three patients achieved good to excellent pain relief that lasted 24-72 hours after each treatment session. CONCLUSION: PENS therapy is a useful supplement to opioid analgesics for the management of pain secondary to bony metastasis in terminal cancer patients. PMID- 9874012 TI - Validity of MMPI-2 profiles in chronic back pain patients: differences in path models of coping and somatization. AB - OBJECTIVE: To show clinical utility and empirical validity of Minnesota Multiphasic Personality Inventory-2 (MMPI-2) chronic pain patient subgroups by identification of differential multivariate relationships across groups. METHOD: This study used structural equation modeling to test cognitive coping strategies and somatization as mediator variables in path models with pain severity and depression used as exogenous (independent) variables and patient's activity level as the final endogenous (dependent) variable, across MMPI-2 profiles. RESULTS: Hierarchical cluster analysis, performed on a sample of 569 chronic low back patients, resulted in four cluster profiles identifiable as those found in previous work with the MMPI-2 (within normal limits, V-type, neurotic triad, and depressed-pathological). Somatization mediated the relationship between depression and activity level for the neurotic triad group but not the other three groups. A positive linear relationship was found between somatization and depression for the within normal limits, neurotic triad, and depressed pathological groups, whereas their linear association was negative for the V-type group. Cognitive coping strategies mediated the relationship between depression and activity level for the within normal limits group. In addition, cognitive coping was predictive of activity level for the within normal limits, V-type, and neurotic triad groups but not for the depressed-pathological group. CONCLUSION: Consistent with previous cluster analytic studies, this study replicated four MMPI-2 cluster profile groups in chronic pain patients. These results have also shown that several multivariate relationships between variables are different across MMPI-2 groups, providing evidence for the validity for these MMPI-2 subgroups. PMID- 9874013 TI - Agreement between child and parent reports of pain. AB - OBJECTIVE: Parents are often the primary source of information regarding their children's pain in both research and clinical practice. However, parent-child agreement on pain ratings has not been well established. The objective of the present study was to examine agreement between child- and parent-rated pain following minor surgery. SETTING: Tertiary care children's hospital. PARTICIPANTS: A total of 110 children (56.4% male) aged 7-12 years undergoing surgery and their parents. OUTCOME MEASURES: Parents and children independently rated pain intensity by using a 7-point Faces Pain Scale on the day of the child's surgery and the following 2 days. RESULTS: Correlations (both Pearson's and intraclass correlation coefficients) indicated a highly significant relationship between child and parent ratings. However, kappa statistics indicated only poor to fair agreement beyond chance. Parents tended to underestimate their children's pain on the day of surgery and the following day, but not on the second day following surgery. When children's and parents' pain ratings for each of the 3 days were collapsed into a no-pain/low-pain group or a clinically significant pain group, kappa statistics indicated fair to good agreement. Parents demonstrated low levels of sensitivity in identifying when their children were experiencing clinically significant pain. CONCLUSIONS: Correlations between parent and child pain reports do not accurately represent the relationship between these ratings and in fact overestimate the strength of the relationship. Parents' underestimation of their child's pain may contribute to inadequate pain control. PMID- 9874014 TI - Assessment of pain during medical procedures: a comparison of three scales. AB - OBJECTIVE: Pain assessment is crucial to pain research. Knowledge about the strengths and weaknesses of pain measures is important to the continued advancement of our understanding of pain. The purpose of the present study was to compare the validity and utility of three measures of pain intensity during a medical procedure known to produce pain: an abortion. DESIGN: Assignment to one of three pain intensity assessment instruments, which were subsequently used to assess pain during an abortion procedure. Comparison of the relative sensitivity of the measures to assess changes in pain using a series of repeated measures analyses of variance. The relative utility of the measures was compared by examining the rates of accurate responses to each. SUBJECTS: Fifty-eight women presenting for a first-trimester abortion. MEASURES: Visual analog scale (VAS), the verbal 11-point Box Scale (Verbal BS-11), and the 21-point Box Scale (BS-21). RESULTS: All three pain intensity measures detected changes in pain during the abortion procedure. Rates of incorrect responses were higher for the Verbal BS-11 and the VAS than for the BS-21. CONCLUSIONS: The results supported the validity of each of the three measures used, although some superiority for the BS-21 over the Verbal BS- 11 and VAS exists. Patients had some difficulty completing the paper-and-pencil VAS during the procedure. In addition and consistent with previous research, some patients treated the Verbal BS-11 as a 21-point scale by responding with numbers between two whole numbers on the 0-10 measure. Overall, practical issues led us to conclude that the BS-21 is an excellent choice for assessing real-time abortion pain. PMID- 9874015 TI - Oral methadone for the treatment of severe pain in hospitalized children: a report of five cases. AB - OBJECTIVE: Pain relief is still inadequate in many hospitalized patients, especially children in whom suboptimal use of analgesic drugs is still common. In the past 2 years, oral methadone has been used extensively in our institution for treating children with persistent pain from cancer, burns, or trauma who were capable of oral intake and whose pain was not relieved by nonopioid medications. SETTING: Tertiary university hospital. PATIENTS: Of the 70 children treated thus far with oral methadone, five are described in the present report. MAIN OUTCOME MEASURE: Pain relief, acceptability, and side effects of oral methadone in children with pain. RESULTS: Treatment with oral methadone (0.1% in 10% glucose, dose range of 0.2-0.6 mg/kg/day) for time periods of up to 6 weeks resulted in a rapid onset and stable pain relief, with no major side effects. No adverse responses were encountered after discontinuation of treatment. In three of the children, a parent-controlled analgesia regimen was successfully employed. CONCLUSIONS: Oral methadone can be recommended for babies and children who have severe pain that is not alleviated by nonopioid medications and who are capable of oral intake. PMID- 9874016 TI - Gabapentin for treatment of neuropathic pain in a 12-year-old girl. AB - We report on a 12-year-old girl with postthoracotomy neuropathic pain. A variety of treatments for the pain were ineffective. The symptoms resolved following the institution of therapy with gabapentin. PMID- 9874017 TI - Contact dermatitis in children: 6 years experience (1992-1997). AB - We report the results of patch tests carried out over the last 6 years (1992 1997) on 141 children. This corresponds to 2.8% of the total patients who were patch tested in our Allergy Department (5,014). These 141 children were patch tested using the TRUE Test standard series and an additional allergen, metallic mercury. Allergens from the Chemotechnique medicaments, cosmetics, plants and shoe series were added in some. The most frequent localization of eczema was to the feet, face and hands and the most frequent suspected causes were costume jewelry, footwear, topical medicaments and cosmetics. The most frequent allergens, assessed by degree of relevance, were first, nickel, followed by cobalt, mercurials (thimerosal and metallic mercury), fragrance and rubber chemicals (naphthyl mix, mercapto mix, carba mix and PPD mix). 13 children, 18% of the total, between the ages of 12 and 14 years, showed positivities relevant to some occupation, i.e., metalworking, building, hairdressing, catering, etc. Once patch tests had been carried out and relevant positive patch tests assessed, the most frequent diagnoses were contact dermatitis 45%, atopic dermatitis 25%, juvenile plantar dermatitis 15%, and other diagnoses in the remaining 15%. PMID- 9874019 TI - Oral and epicutaneous desensitization in urushiol contact dermatitis in guinea pigs sensitized by 2 methods of different sensitizing potency. AB - This investigation was conducted to study the relationship between the different intensities of sensitization to urushiol induced by the maximization test method (MT) or by the ear-flank method (EF) in guinea pigs, and hyposensitization induced by repeated oral and/or epicutaneous administration of the allergen, and to compare the immune tolerance achieved between these routes of dosing. During the observation period of 10 weeks, both oral and epicutaneous administration produced hyposensitization to urushiol after weak sensitization by EF; however, only combined administrations via the oral and epicutaneous routes achieved hyposensitization after potent sensitization, while inadequate tolerance was induced by oral dosing and none by epicutaneous dosing. Thus, it is indicated that difficulty in establishing immune tolerance may increase with increasing intensity of primary sensitization, and that hyposensitizing potency tends to increase in the order epicutaneous, oral and combined (e.p.+p.o.) administrations of urushiol. PMID- 9874018 TI - Metal release from gold-containing jewelry materials: no gold release detected. AB - Metal release from 13 different gold-containing jewelry alloys stored for 1 and 3 weeks in artificial sweat was analysed. For chemical analysis, inductively coupled plasma detection (ICP) and atomic absorption spectrophotometry (AA), with flame and with furnace, were used. No release of gold was detected. It is unlikely that skin contact with gold-containing metallic items such as jewelry is responsible for inducing contact allergy to gold or allergic contact dermatitis due to the gold. The patch-test reactivity to gold sodium thiosulfate needs to be explained by some other mechanism(s). PMID- 9874020 TI - Fragrances and other materials in deodorants: search for potentially sensitizing molecules using combined GC-MS and structure activity relationship (SAR) analysis. AB - Deodorants are one of the most frequently-used types of cosmetics and are a source of allergic contact dermatitis. Therefore, a gas chromatography - mass spectrometric analysis of 71 deodorants was performed for identification of fragrance and non-fragrance materials present in marketed deodorants. Futhermore, the sensitizing potential of these molecules was evaluated using structure activity relationships (SARs) analysis. This was based on the presence of 1 or more chemically reactive site(s), in the chemical structure, associated with sensitizing potential. Among the many different substances used to formulate cosmetic products (over 3500), 226 chemicals were identified in a sample of 71 deodorants. 84 molecules were found to contain at least 1 structural alert, and 70 to belong to, or be susceptible to being metabolized into, the chemical group of aldehydes, ketones and alpha,beta-unsaturated aldehydes, ketone or esters. The combination of GC-MS and SARs analysis could be helpful in the selection of substances for supplementary investigations regarding sensitizing properties. Thus, it may be a valuable tool in the management of contact allergy to deodorants and for producing new deodorants with decreased propensity to cause contact allergy. PMID- 9874021 TI - Is benzoquinone the prohapten in cross-sensitivity among aminobenzene compounds? AB - Cross-sensitivity in allergic contact dermatitis is a simultaneous allergy to 2 or more contact substances which have in common an antigenic determinant or a metabolic derivative. One of the most notable examples is the cross-sensitivity among aromatic compounds which may be oxidized in vivo to benzoquinone (BQ). However, it has also been hypothesized that the allergenicity and cross sensitization are modulated by the chemical reactivity of the substituents in the para position. A serial dilution of BQ (from 1% to 0.1% in pet.) and three 1,4 substituted benzene derivatives (p-aminophenol, hydroquinone, metol), theoretically capable of conversion to BQ by oxidation, were patch tested in 22 p phenylenediamine (PPD) positive patients and in 20 controls. The patients and a further 116 subjects with a positive history of sensitivity to 1 or more aminoaromatic compounds were also tested with some haptens of the para group (PPD, p-aminobenzoic acid, p-aminodiphenylamine, benzocaine, procaine chloride, p toluenediamine sulfate). The results show that (i) the optimal patch test concentration for BQ was 0.2%, (ii) only 4 of the 22 patients allergic to PPD gave a clearly positive allergic reaction to BQ, and (iii) the number of positive reactions to the aromatic compounds was correlated with the presence of activating (-NH2, -OH, -CH3) and deactivating (-COOH) groups in the para position or, perhaps, with their effect on percutaneous penetration. The data suggest that BQ is not the only intermediate in the cross-sensitization of para group haptens. This is probably conditioned by other oxidation products and/or the chemical structure of the substituents in position 4 of the benzene ring. PMID- 9874022 TI - Development of occupational skin diseases during vocational training in baker and confectioner apprentices: a follow-up study. AB - Baker and confectioner apprentices have been included in a prospective follow-up study in the region of East Thuringia, to evaluate risk factors for the development of hand dermatitis. Starting in August 1996, the apprentices have been interviewed and examined in a standardized way right at the beginning (n=91) of their vocational training, 6 (n=79) and 12 months (n=63) later. To gain objective data, TEWL score was also assessed at 3 standardized sites on the hands. In their case histories, 3.3% (n=3) of the apprentices reported hand dermatitis. The 1st assessment after 2 to 4 weeks vocational training revealed hand eczema in 17.5% (n=16) of the individuals. At the follow-up after 6 months of training 29.1% (n=23), and after 12 month of training 27.0% (n=17), of the apprentices had hand dermatitis of mild to moderate severity. Within 6 months, the TEWL score rose significantly (p<0.001) from 11.9 g m(-2) h(-1) (-/+5.4) to 16.8 g m(-2) h(-1) (-/+9.5). After 12 months, the TEWL score was 14.9 g m(-2) h( 1) (-/+4.6). For the atopic skin diathesis (>10 points of Diepgen's atopy score), there was a significant tendency to be a predictive factor for the development of occupational hand dermatitis. TEWL score failed to correlate with the development of skin damage in our study. The same was true for respiratory atopy and metal sensitization. PMID- 9874023 TI - Eyelid dermatitis from methacrylates used for nail enhancement. PMID- 9874024 TI - Immediate hypersensitivity to sesame in foods and cosmetics. PMID- 9874025 TI - Active sensitization to (meth)acrylates. PMID- 9874026 TI - Textile dye contact dermatitis in an allergic population. PMID- 9874027 TI - Skin allergy from exposure to alloys of chromium. PMID- 9874028 TI - Cutaneous reaction to captopril with positive patch test and lack of cross sensitivity to enalapril and benazepril. PMID- 9874029 TI - Pseudoinoculum barbae: beard-induced contact dermatitis. PMID- 9874030 TI - The seasonal influence on patch test reactions is significant in Israel. PMID- 9874031 TI - Difficulties in the interpretation of patch test reactions to ophthalmic beta blockers. PMID- 9874032 TI - The patch test concentration for methylchloroisothiazolinone/methylisothiazolinone. PMID- 9874033 TI - Contact allergy to 2-aminoethanol (monoethanolamine) in a soluble oil. PMID- 9874034 TI - Occupational contact sensitivity to aluminium in a machine construction plant worker. PMID- 9874035 TI - Allergic contact dermatitis due to diclofenac and indomethacin. PMID- 9874036 TI - Hand dermatitis caused by ferns. PMID- 9874037 TI - Contact dermatitis from dipivefrine with possible cross-reaction to epinephrine. PMID- 9874038 TI - Nickel in tap water in Warsaw. PMID- 9874039 TI - The role of coumarin in patch testing. PMID- 9874040 TI - Occupational contact sensitization to corticosteroids. PMID- 9874041 TI - Patch testing with maleopimaric acid in an occupational dermatology clinic. PMID- 9874042 TI - Interactions of divalent ions in nickel-sensitive subjects. PMID- 9874043 TI - Direct stent implantation without predilatation using the MultiLink stent. AB - The standard coronary stent implantation technique requires routine predilatation of the target lesion with a balloon catheter. In this study, we prospectively studied the feasibility and efficiency of elective coronary stent implantation without predilatation. In 94 patients who presented with various ischemic syndromes, direct implantation of 100 balloon expandable ACS MultiLink stents (7 over-the-wire, 93 rapid exchange) was attempted in 100 coronary lesions selected to have favorable characteristics. The stent crossed the lesion without predilatation in 97 cases (97%) and was successfully deployed in 93 (95.8%). In 4 patients, adjunctive high-pressure postdilatation was necessary to achieve optimal stent expansion. Reference vessel diameter was 3.12+/-0.77 mm and lesion length 8.8+/-2.7 mm. Minimal luminal diameter increased from 0.95+/-0.38 mm to 2.98+/-0.28 mm and diameter stenosis decreased from 71+/-11% to 8+/-11% after stenting. One occlusive dissection was treated by a second stent. There were no major in-hospital complications. At 1 month follow-up, 1 subacute thrombotic occlusion occurred. These results indicate that in a carefully selected coronary lesion subset, elective stent implantation without predilatation can be safely and effectively performed. The long-term results of this approach and possible advantages over the conventional implantation techniques remain unclear and need to be evaluated in further clinical studies. PMID- 9874044 TI - Impact of an aggressive coronary stenting strategy on the incidence of target lesion revascularization. AB - Coronary stenting has been shown to reduce the incidence of target lesion revascularization (TLR) compared with balloon angioplasty in highly selected patients. However, the impact of an aggressive coronary stenting strategy in unselected patients on the overall incidence of TLR is unclear. We assessed the effect of increased stenting by comparing long-term results in consecutive patients who underwent successful percutaneous revascularization (with or without stents) during June to December 1995 (n=347) with those in June to December 1996 (n=401). Stents were used in 22.5% of patients in 1995 versus 66.1% in 1996 (p <0.0001). Mean age of the patients was 62+/-11 years (71% men) in 1995 versus 63+/-10 in 1996 (76% men) (p=NS). The 2 groups were well matched with the exception that the 1996 cohort included more patients with unstable coronary syndromes (25% in 1995 vs 34% in 1996 (p=0.003). There was no significant difference in the incidence of in-hospital adverse events. After 12 months of follow-up (complete in 95% of patients in each group), the incidence of TLR was significantly lower in the 1996 cohort than in the 1995 cohort (8.5% vs 14.7%, p=0.0075). This was mainly due to reduced requirement for repeat angioplasty in 1996 patients compared with 1995 (6.5% vs 11.8%, p=0.011). It is concluded that in an unselected patient population, an aggressive coronary stenting strategy was associated with a marked overall reduction in requirement for TLR over a 12-month period. PMID- 9874045 TI - Two- to three-year follow-up of patients with single-vessel coronary artery disease randomized to PTCA or medical therapy (results of a VA cooperative study). Veterans Affairs Cooperative Studies Program ACME Investigators. Angioplasty Compared to Medicine. AB - Despite increasing use of percutaneous transluminal coronary angioplasty (PTCA) to treat stenotic coronary artery disease, there are relatively few prospective studies evaluating its long-term effectiveness. We prospectively randomized 212 stable patients with provocable myocardial ischemia and single-vessel subocclusive coronary disease to receive primary therapy with either PTCA or medical therapy. This report presents the clinical follow-up of these patients at a mean, after randomization, of 2.4 years for interview and 3.0 years for exercise testing. Of the 212 patients originally randomized, 175 received an extended follow-up interview, and 132 underwent exercise testing; 62% of patients in the PTCA group were angina free compared with 47% of patients in the medical group (p <0.05). Furthermore, exercise duration as measured by treadmill testing was prolonged by 1.33 minutes over baseline in the PTCA group, whereas it decreased by 0.28 minutes in the medical group (p <0.04). Although the angina free time on the treadmill was not different (p=0.50), fewer patients in the medical group developed angina on the treadmill at 3 years than those in the PTCA group (p=0.04). By 36 months, excluding the initial randomized PTCA, use of PTCA and use of coronary artery bypass surgery were not different in the 2 treatment groups. These data indicate that some of the early benefits derived from PTCA in patients with single-vessel coronary artery disease are sustained, making it an attractive therapeutic option for these patients. PMID- 9874046 TI - Increased intensity of contrast material immediately after late angioplasty of infarct-related coronary artery is associated with reduced ventricular volumes at six months. AB - To assess the contribution of residual muscle perfusion in the infarcted territory to prevent ventricular remodeling, 24 patients with 1-vessel disease underwent coronary angiography and angioplasty of a critical left anterior descending coronary stenosis 18+/-11 days after a first anterior myocardial infarction. The degree of stenosis was assessed using biplane quantitative angiography, whereas ventricular volumes, together with regional wall motion, were computed from single-plane ventriculography. Patients were reevaluated at 6 months after they had been subdivided according to the videointensity of the territory of the culprit vessel, as assessed from images obtained during main stem dye contrast injections before and immediately after angioplasty using a subtraction technique (group A, increased intensity [n= 15]; group B, no change [n=9]), assuming that higher peak intensities reflect greater myocardial blood volume. There was a significant time group interaction for ventricular volumes (diastolic, -13+/-12% for group A vs +20+/-24% for group B, p=0.008; systolic, 15+/-19% for group A vs +18+/-36% for group B, p=0.017), although no interaction was evident for the degree of resolution of coronary stenosis or the extent of recovery of regional dysfunction. The effects on volumes were paralleled by changes in ventricular end-diastolic pressure (-3+/-7 mm Hg in group A vs +5+/-6 mm Hg in group B, p=0.006), although baseline clinical characteristics and medical regimen over the 6-month period were quite comparable between the 2 groups. In conclusion, despite late angioplasty of the culprit vessel, ventricular remodeling is prevented mainly when the procedure guarantees improved perfusion at the muscular level. The result is not necessarily mediated by recovery of regional systolic function. PMID- 9874047 TI - Heterogeneous fate of perfusion and contraction after anterior wall acute myocardial infarction and effects on left ventricular remodeling. AB - After acute myocardial infarction, patency of infarct vessel and extent of left venticular (LV) dysfunction are major determinants of ventricular remodeling. Spontaneous, delayed reperfusion in the infarct zone occurs in a sizeable number of patients well after the subacute phase. The aim of this study was to determine the relation between the occurrence of this spontaneous, delayed reperfusion and LV remodeling. In 84 patients, resting LV volumes, topography, regional function, and perfusion were quantitatively evaluated by 2-dimensional echocardiography and sestamibi tomography 5 weeks (study 1) and 7 months (study 2) after anterior Q wave infarction. At study 2, LV end-diastolic volume increased by > 15% in 17 patients (20%, LV remodeling); they had already had at study 1 significantly larger LV volumes, more severe hypoperfusion and wall motion abnormalities, and greater regional dilation than patients with stable LV volumes. Delayed reperfusion occurred in 8 of 17 patients with and in 42 of 67 patients without LV remodeling (47% vs 63%; p=NS). At study 2, LV regional dilation and end-diastolic volumes were stable in patients with, but increased in patients without, spontaneous reperfusion (from 25+/-24% to 29+/-26% at study 2 [p<0.05] and from 65+/-14 to 68+/-18 ml/m2 [p <0.05]). At multivariate analysis, however, regional ventricular dilation at study 1 was the sole predictor of further LV remodeling. Thus, after acute myocardial infarction, spontaneous reperfusion occurring after 5 weeks plays only a minor role in influencing LV remodeling. Benefits from delayed reperfusion seem limited to patients with preserved LV volumes; patients with an enlarged left ventricle 5 weeks after acute infarction are prone to further LV remodeling, irrespective of delayed reperfusion. PMID- 9874048 TI - Effect of intravenous nitroglycerin on lipid peroxidation after thrombolytic therapy for acute myocardial infarction. AB - Free oxygen radicals are produced after coronary artery occlusion and reperfusion. Polyunsaturated fatty acids are oxidized by free radicals to lipid peroxides. Measurements of plasma malondialdehyde (MDA) formed by the breakdown of lipid peroxides are often used as markers of lipid peroxidation. The effect of intravenous nitroglycerin on plasma MDA levels was studied in 43 patients who received thrombolytic therapy for acute myocardial infarction. Plasma MDA levels in patients were elevated on admission to the hospital compared with healthy controls, and normalized within 48 hours. A greater increase in plasma MDA concentrations after thrombolysis was found in patients with noninvasive signs of reperfusion than in patients judged to have a persistent occlusion. In the 23 patients receiving immediate intravenous nitroglycerin infusion, plasma MDA levels did not change from baseline to 90 minutes (0.92+/-0.22 and 0.92+/-0.23 micromol/L, p=0.99), whereas a significant increase was found in the 20 control patients who did not receive nitroglycerin (from 0.83+/-0.22 to 1.01+/-0.30 micromol/L, p=0.0004) (p=0.036 for the difference between groups). Successful reperfusion after thrombolytic therapy entails increased lipid peroxidation. Intravenous nitroglycerin reduces lipid peroxidation during myocardial ischemia and reperfusion. PMID- 9874049 TI - Comparison of dipyridamole stress echocardiography and perfusion scintigraphy for cardiac risk stratification in vascular surgery patients. AB - Dipyridamole single-photon emission computed tomography (SPECT) has a high negative predictive value for perioperative cardiac events, but events are infrequent in patients with a positive test. In contrast, dipyridamole echocardiography is more selective for detection of multivessel disease and thus may have a greater specificity for cardiac events. We therefore compared the ability of dipyridamole SPECT and echocardiography to predict perioperative and long-term cardiac events in 133 patients referred for vascular surgery. The group was also evaluated based on clinical features and ejection fraction. Four patients had surgery cancelled because of high risk and were excluded from further analysis. Among the 129 remaining patients, 21 had coronary revascularization (n=12) or an early cardiac end point (n=9). The sensitivity of SPECT for the prediction of early events (90%) was not significantly different from that of echocardiography (66%, p=NS). The specificity of SPECT (68%) was less than that of echocardiography (88%, p <0.001%), as was the accuracy (72% vs 84%, p=0.02). These findings were replicated after exclusion of patients with treatment end points. During long-term follow-up, 12 patients experienced > or = 1 event: 6 died from cardiac causes, 4 underwent revascularization, and 3 had myocardial infarction. Thus, the specificity of SPECT and echocardiography for late events were 58% and 80%, respectively (p <0.001). The 3-year survival of patients without ischemia during echocardiography or at SPECT was not different (93% vs 94%, p=NS). PMID- 9874050 TI - Diagnosis of coronary vasospasm in patients with clinical presentation of unstable angina pectoris using ergonovine echocardiography. AB - Although coronary vasospasm can contribute to the development of unstable angina, the definite diagnostic method has not been established. The purpose of this study was to determine if ergonovine echocardiography (detection of regional wall motion abnormality during bedside ergonovine challenge) after angiographic confirmation of insignificant fixed disease would be useful and safe in detecting coronary vasospasm in patients with unstable angina. After control of chest pain with medications in patients admitted to the coronary care unit under the tentative diagnosis of unstable angina, diagnostic coronary angiography was performed. All patients with normal or insignificant fixed disease underwent ergonovine echocardiography after discontinuation of medications for 4+/-1 days. Among 208 consecutive patients enrolled for this study, 75% (156 of 208) showed significant fixed disease in the angiography. Ergonovine echocardiography was performed in 52 patients with insignificant disease, and coronary vasospasm was documented in 33 (63%, 33 of 52). No serious procedure-related arrhythmia or myocardial infarction occurred. Esophageal motility disorder and hypertrophic cardiomyopathy were diagnosed in 6 and 3 patients, respectively. Chest pain of undetermined etiology was the final diagnosis at discharge in 10 patients (5%, 10 of 208); among them chest pain redeveloped in 2 patients, and repeated ergonovine echocardiography revealed positive results. Our data suggest that among patients with the clinical presentation of unstable angina, coronary vasospasm is the main cause of myocardial ischemia in a considerable number of patients with a normal or near-normal angiogram, and ergonovine echocardiography after confirmation of absence of significant fixed disease is useful and safe for noninvasive diagnosis of coronary vasospasm in this setting. PMID- 9874051 TI - Chlamydia pneumoniae infection is frequent but not associated with coronary arteriosclerosis in cardiac transplant recipients. AB - We sought to explore the relation between Chlamydia pneumoniae, cytomegalovirus (CMV), and cardiac transplant-associated arteriosclerosis. Serologic evidence of past Chlamydia pneumoniae infection was investigated in 3 patient groups at the time of cardiac catheterization: cardiac transplant recipients (n=49), patients having coronary artery bypass grafting (CABG) (n=39), and a control group free of angiographic coronary artery disease (n=21). High Chlamydia pneumoniae immunoglobulin G titers (> or =1:160) were more frequently observed in cardiac transplant recipients (odds ratio[OR] 13.7; 95% confidence intervals [CI] 1.6 to 117.4, p <0.05) and CABG patients (OR 21.7; 95% CI 1.6 to 287.0, p <0.05) than in controls. However, high Chlamydia pneumoniae titers did not distinguish between cardiac transplant recipients with or without angiographic transplant-associated arteriosclerosis or CABG patients with or without bypass vein graft disease. Furthermore, there was no significant relation between elevated Chlamydia pneumoniae titers and the presence or progression of transplant-associated arteriosclerosis in the subgroup of patients who were also CMV positive. Yet, analysis of the same angiograms demonstrated an association between CMV infection and the recent progression of transplant-associated arteriosclerosis. Thus, patients with cardiac transplantation have evidence of past Chlamydia pneumoniae and CMV infection but Chlamydia pneumoniae does not appear to have an independent role or synergistic relation to CMV in the development of transplant-associated arteriosclerosis. PMID- 9874052 TI - Effects of pravastatin on thoracic aortic atherosclerosis in patients with heterozygous familial hypercholesterolemia. AB - Data regarding the effects of plasma lipid lowering on the evolution of thoracic aortic atherosclerosis (TAA) are scarce. In this study, we performed transesophageal echocardiography to characterize TAA in 16 newly diagnosed patients with heterozygous familial hypercholesterolemia and to follow its evolution after 2 years of statin treatment. TAA was graded as follows: grade I = normal intima; grade II = increased intimal echo density without thickening; grade IIIA = increased intimal echo density with single atheromatous plaque < or = 3 mm; grade IIIB = multiple plaques < or = 3mm; grade IV = > or = 1 plaque >3 mm; and grade V = mobile or ulcerated plaques. Baseline aortic intimal morphology was grade I in one patient, grade II in 4, grade IIIA in 6, grade IIIB in 3, and grade IV in 2 patients. Hypolipidemic treatment resulted in significant reductions in plasma total cholesterol and low-density lipoprotein (LDL) cholesterol. Follow-up aortic morphology was grade I in 5 patients, grade II in 2, grade IIIA in 3, grade IIIB in 3, and grade IV in 3 patients. TAA remained stable in 7 patients, progressed in 3, and regressed in 6 patients. TAA evolved in a uniform manner in the ascending aorta, aortic arch, and descending aorta. Patients with TAA regression were younger (39+/-14 vs 52+/-8 years, p=0.038) and had a greater decrease in plasma LDL cholesterol as a result of treatment (138+/ 56 vs 73+/-55 mg/dl, p=0.036) than patients with TAA stability or progression. These observations support the hypothesis that hypolipidemic treatment may favorably affect the course of TAA in patients with heterozygous familial hypercholesterolemia. PMID- 9874053 TI - Long-term efficacy of low-density lipoprotein apheresis on coronary heart disease in familial hypercholesterolemia. Hokuriku-FH-LDL-Apheresis Study Group. AB - Familial hypercholesterolemia (FH) is characterized by severe hypercholesterolemia and premature coronary heart disease (CHD). The lower the plasma cholesterol level, the more likely it is that CHD can be prevented or retarded; aggressive cholesterol-lowering therapies may be indicated for FH patients with CHD. This study describes the long-term (6 years) safety and efficacy of intensive cholesterol-lowering therapies with low-density lipoprotein (LDL) apheresis in heterozygous FH patients with CHD. One hundred thirty heterozygous FH patients with CHD documented by coronary angiography had been treated by cholesterol-lowering drug therapy alone (n=87) or LDL apheresis combined with cholesterol-lowering drugs (n=43). Serum lipid levels and outcomes in each treatment group were compared after approximately 6 years. Both treatment groups had significant reductions in serum cholesterol, LDL cholesterol, and high density lipoprotein cholesterol levels. LDL apheresis significantly reduced LDL cholesterol levels from 7.42+/-1.73 to 3.13+/-0.80 mmol/L (58%) compared with group taking drug therapy, from 6.03+/-1.32 to 4.32+/-1.53 mmol/L (28%). With Kaplan-Meier analyses of the coronary events including nonfatal myocardial infarction, percutaneous transluminal coronary angioplasty, coronary artery bypass grafting, and death from CHD, the rate of total coronary events was 72% lower in the LDL-apheresis group (10%) than in drug therapy group (36%) (p=0.0088). It is concluded that LDL-apheresis is effective as treatment of CHD in FH heterozygotes, and may become the therapy of choice in severe types of FH. PMID- 9874054 TI - Influence of heart rate on stroke volume variability in atrial fibrillation in patients with normal and impaired left ventricular function. AB - Both resting tachycardia and irregular ventricular rhythm may contribute to impaired cardiac performance in atrial fibrillation (AF). This study assesses the relation between resting heart rate and beat-to-beat changes in left ventricular (LV) ejection and filling in patients with normal and impaired LV systolic function. Beat-to-beat variation in LV outflow and inflow velocity-time integral was measured using pulsed Doppler ultrasound in 39 patients with chronic AF and normal (n=22) or impaired (n=17) LV systolic function. Aortic velocity-time integral variability increased with mean heart rate (p=0.003) even though RR interval variability decreased (p <0.001). Aortic velocity-time integral was more sensitive to the duration of both the preceding (p <0.001) and prepreceding (p <0.001) RR intervals at higher heart rates. These relations were similar for patients with normal and impaired LV systolic function. The sensitivity of the filling velocity-time integral to RR interval variability also increased with heart rate (p <0.001). However, at higher heart rates the filling velocity-time integral (p=0.009) and filling time (p=0.005) were less sensitive to change in RR intervals in patients with impaired LV function. We conclude that beat-to-beat stroke volume variability in AF increases with heart rate. Stroke volume variability was not influenced by LV systolic function. PMID- 9874055 TI - Does withdrawal of antihypertensive medication increase the risk of cardiovascular events? Trial of Nonpharmacologic Interventions in the Elderly (TONE) Cooperative Research Group. AB - The Fifth Report of the Joint National Committee on Detection, Evaluation, and Treatment of High Blood Pressure recommends that attempts to discontinue antihypertensive drug therapy be considered after blood pressure (BP) has been controlled for 1 year. However, discontinuation of drug therapy could unmask underlying conditions and precipitate clinical cardiovascular events. The Trial of Nonpharmacologic Interventions in the Elderly (TONE) was a clinical trial of the efficacy of weight loss and/or sodium reduction in controlling BP after withdrawal of drug therapy in patients with a BP< 145/85 mm Hg on 1 antihypertensive medication. Of 975 participants, 886 entered the drug withdrawal phase of the trial and 774 were successfully withdrawn from their medications. Thirty-three events (stroke, transient ischemic attack, myocardial infarction, arrhythmia, congestive heart failure, angina, other) occurred between randomization and the onset of drug withdrawal (median time 3.6 months), 57 events occurred either during or after drug withdrawal (14.0 months), and 36 events occurred after resumption of antihypertensive therapy (15.9 months). Event rates per 100 person-years were 5.5, 5.5, and 6.8 for the 3 time periods (p=0.84) in the nonoverweight group and 7.2, 5.2, and 5.6 (p=0.08) in the overweight group. The study shows that antihypertensive medication can be safely withdrawn in older persons without clinical evidence of cardiovascular disease who do not have diastolic pressure > or = 150/90 mm Hg at withdrawal, providing that good BP control can be maintained with nonpharmacologic therapy. PMID- 9874056 TI - Early expression of a malignant phenotype of familial hypertrophic cardiomyopathy associated with a Gly716Arg myosin heavy chain mutation in a Korean family. AB - The clinical course and prognosis of familial hypertrophic cardiomyopathy (HCM) are different according to the type of mutation in the genes for sarcomere proteins. It has been disputed that a mutation, which occurs at a functionally important region in the sarcomere proteins, may increase the penetrance and expressivity of the disease. We searched for a causative mutation in an HCM family, which is characterized by early expression of clinical phenotype, high incidence of sudden death at young ages, and progressive heart failure in adults. Among the 32 family members in 4 generations, 13 were affected; 4 died suddenly before age 16, 2 children have already had full expression of the cardiac hypertrophy, and other adults have either progressive heart failure or poor left ventricular systolic functions. PCR-SSCP (polymerase chain reaction-single strand confirmation polymorphism) analysis of genomic DNAs isolated from peripheral blood leukocytes of the family members identified a Gly716Arg mutation in the cardiac beta-myosin heavy chain gene, which was cosegregated with the clinical phenotype. The mutation is localized near a functionally important site of the myosin heavy chain, the 2 active thiols, which contribute to the adenosine triphosphatase activity of myosin S1. This family provides further evidence that the mutation, which occurs at a functionally important site of the myosin heavy chain, is associated with the high penetrance and early expression of HCM. PMID- 9874057 TI - QT dispersion and risk factors for sudden cardiac death in patients with hypertrophic cardiomyopathy. AB - This study examines the relation of QT dispersion (QTd) on a surface electrocardiogram (ECG) to clinical features and established risk factors of sudden cardiac death (SCD) in patients with hypertrophic cardiomyopathy (HC). One hundred fifty-six consecutive patients with HC (91 men, mean age 41+/-15 years, range 7 to 79) and 72 normal subjects (41 men, mean age 39+/-9 years, range 20 to 60) were studied. Standard 12-lead ECGs were recorded from each subject using a MAC VU electrocardiograph. Patients with nonsinus rhythm, atrioventricular conduction block, QRS duration > 120 ms, age < 15 years, and low amplitude T waves were excluded from the analysis (n=51). Another 22 patients who were receiving amiodarone and/or sotalol therapy were also excluded. QT interval and QTd were measured using automated analysis in the remaining 83 patients (46 men, age 40+/-14 years, range 16 to 76). QT interval (406+/-38 ms), QTc interval (432+/-27 ms), and QTd (43+/-25 ms) were significantly greater in patients with HC than in normal controls (386+/-31 ms, 404+/-16 ms, 26+/-16 ms, respectively) (p <0.0001). QTd was significantly greater in patients with HC with chest pain compared with asymptomatic or mildly symptomatic patients (50+/-28 ms vs 37+/-20 ms, p=0.02). Increased QTd was found in patients with dyspnea New York Heart Association functional classes II/III than in those with dyspnea New York Heart Association functional class I (50+/-27 ms vs 38+/-22 ms, p=0.04). QTd was weakly correlated with maximum left ventricular wall thickness (r=0.228, p=0.038). No significant association was found between QTd and any risk factors for SCD. Thus, patients with HC have increased QTd. The QTd correlates with symptomatic status. Assessment of QTd might provide complementary clinical characterization of patients with HC but its relation to SCD remains uncertain. PMID- 9874058 TI - Standardized guidelines for the interpretation of dobutamine echocardiography reduce interinstitutional variance in interpretation. AB - Subjective interpretation of dobutamine echocardiograms provides only moderate interinstitutional observer agreement if nonunified data acquisition and assessment criteria are applied. The present study was undertaken to evaluate parameters associated with low interinstitutional observer agreement in the interpretation of dobutamine echocardiograms and to analyze whether standardized interpretation criteria improve interinstitutional observer agreement. One hundred fifty dobutamine echocardiograms (dobutamine up to 40 microg/kg/min body weight and atropine up to 1 mg) were evaluated at 5 centers. Clinical, procedural, and echocardiographic parameters were included in the analysis of variables with significant impact on interinstitutional agreement. Standardized interpretative criteria were established, and 90 dobutamine echocardiograms were reanalyzed by 3 observers using a standardized image display. Multivariate analysis demonstrated low image quality (odds ratio [OR] 0.19, 95% confidence interval [CI] 0.08 to 0.45, p=0.0002), low severity of induced wall motion abnormality (OR 0.17, 95% CI 0.07 to 0.40, p <0.0001), and a low peak rate pressure product (OR 0.93, 95% CI 0.43 to 2.27, p=0.0382) to result in a low interinstitutional agreement. Standardization of image display in cine loop format and of dobutamine stress echo interpretation criteria resulted in improvement in test result categorization as normal or abnormal, with a kappa value of 0.50, compared with 0.39 using the original subjective interpretation. In conclusion, image quality, the severity of induced wall motion abnormalities, and the obtained rate-pressure product have a significant impact on the interpretation homogeneity of dobutamine echocardiograms. Standardization of image display in cine loop format and of reading criteria results in improved interinstitutional agreement in interpretation of stress echocardiograms. PMID- 9874059 TI - Importance of exercise capacity in the interpretation of a myocardial ischemic response to exercise testing. Multicenter Study of Myocardial Ischemia Group (MSMI). AB - Patients who were stable 1 to 6 months after a cardiac event underwent routine exercise testing with thallium scintigraphy. The prognosis of patients with good exercise capacity (Bruce stage 3) was similar whether or not ischemia was demonstrated and similar to patients with reduced exercise capacity and no ischemia, whereas the presence of both ischemia and a reduced exercise tolerance identified patients with a significantly poorer prognosis. PMID- 9874060 TI - Dobutamine-induced ST-segment elevation in patients without myocardial infarction. AB - During dobutamine stress echocardiography, ST-segment elevation developed in 20 of 372 patients (5%) without previous myocardial infarction and was associated with a transient severe asynergy of the myocardial region corresponding to the site of ST elevation. In 17 of 19 patients, ST-segment elevation was associated with a critical stenosis of the ischemia-related coronary artery, whereas in 2 of 19 patients with no critical lesions of the ischemia-related artery, coronary vasospasm was the most likely mechanism of myocardial ischemia. PMID- 9874061 TI - Coronary arterial anatomy in patients with left ventricular systolic dysfunction without chest pain or previous myocardial infarction. AB - Patients with left ventricular systolic dysfunction without chest pain or previous myocardial infarction are unlikely to have 3-vessel coronary artery disease and are very unlikely to have coronary anatomy suitable for bypass grafting. Of our 108 subjects, only 3 (3%) had 3-vessel coronary artery disease thought to be suitable for revascularization. PMID- 9874062 TI - Prognostic significance of cerebrovascular disease in 11,526 chronic coronary artery disease patients. Bezafibrate Infarction Prevention (BIP) Study Group. AB - Patients with chronic CAD and a history of cerebrovascular events were compared with patients without prior cerebrovascular events to assess the effect of these events on 5-year prognosis. Despite adjustment for older age and higher comorbidity among patients who had experienced a cerebrovascular event, a history of such an event was associated with an increased risk of 1.86 for total mortality. PMID- 9874063 TI - Endothelium-dependent flow-mediated vasodilation in coronary and brachial arteries in suspected coronary artery disease. AB - Previous studies showed a weak correlation between endothelial function of the coronary arteries as assessed by acetylcholine and brachial artery vasomotion during reactive hyperemia. When the same stimulus was used, we obtained a strong correlation between flow-mediated dilation in the coronary and brachial arteries (r=0.78, p <0.001), so that noninvasive assessment of flow-mediated dilation in the brachial artery could be used as a surrogate measure for coronary artery endothelial function. PMID- 9874065 TI - Modulatory impact of cardiac rehabilitation on hyperhomocysteinemia in patients with coronary artery disease and "normal" lipid levels. AB - In coronary patients with "relatively normal" lipid values and hyperhomocystinemia (levels > or =15 micromol/L), significant 12% reductions in homocysteine levels occurred after cardiac rehabilitation and exercise training. This benefit from cardiac rehabilitation and exercise training may lead to 20% to 30% reductions in overall coronary artery disease risk. PMID- 9874064 TI - Clinical and angiographic results of stenting for long coronary arterial atherosclerotic lesions. AB - A prospective registry of 187 patients who underwent percutaneous coronary angioplasty with attempted long NIR stent delivery was performed. A successful stent delivery was achieved in 93% of cases with a low rate of major cardiovascular events, and 6-month follow-up showed low rates of clinical events, new revascularization procedures, and angiographic restenosis. PMID- 9874066 TI - Timing of thromboembolic events after electrical cardioversion of atrial fibrillation or flutter: a retrospective analysis. AB - Pooled data from 32 studies were reviewed to assess the timing of thrombolic complications after cardioversion of atrial fibrillation or flutter. We found that 98% of embolic episodes occurred within 10 days of cardioversion. PMID- 9874067 TI - Patent ductus arteriosus occlusion using detachable coils. AB - Occlusion of patent ductus arteriosus was performed using detachable coils in 193 cases, with 181 successful implants and a low embolization rate. This technique is safe and effective for occlusion of ductuses of various sizes, and is low cost. PMID- 9874068 TI - Aortic sinotubular junction calcium as a marker of severe aortic atherosclerosis. AB - Because the significance of calcification of the sinotubular junction is poorly understood, we retrospectively investigated its association with aortic atherosclerosis in 101 patients who underwent transesophageal echocardiography. Such calcification was found to be a marker of more severe aortic atherosclerosis and, specifically, aortic arch disease. PMID- 9874069 TI - Correlation of thoracic aortic atherosclerotic plaque detected by multiplane transesophageal echocardiography and cardiovascular risk factors. AB - This study of 416 patients identified age, male gender, smoking, diabetes, hypertension, and hypercholesterolemia as independent predictors of thoracic aortic atherosclerotic plaque. Age, smoking, hypercholesterolemia, hypertension, and diabetes were predictors of the severity and extent of thoracic aortic atherosclerosis. PMID- 9874070 TI - Effect of long-term estrogen therapy on brachial arterial endothelium-dependent vasodilation in women with Raynaud's phenomenon secondary to systemic sclerosis. AB - Endothelium-dependent and endothelium-independent dilation were reduced in patients with Raynaud's phenomenon secondary to systemic sclerosis. Conjugated estrogen given for 4 weeks significantly improved endothelial function compared with placebo. PMID- 9874071 TI - Imaging technique and clinical utility for electrophysiologic procedures of lower frequency (9 MHz) intracardiac echocardiography. AB - Intracardiac echocardiography using a new 9-MHz ultrasound catheter was performed in 30 patients undergoing percutaneous catheter mapping and radiofrequency ablation of a tachyarrhythmia, because the imaging capabilities with this intracardiac echocardiographic catheter permit detailed identification of normal and abnormal cardiac anatomy with improved imaging depth. Intracardiac echocardiography is of significant clinical utility during ablation for guiding interatrial septal puncture, assessing placement and contact of mapping/ablation catheters, monitoring ablation lesion morphologic changes, and diagnosing procedure-related complications. PMID- 9874072 TI - Role of apical aneurysm on the prognosis of chronic Chagas' disease. PMID- 9874073 TI - Optimal imaging in assessment of right ventricular function in tetralogy of Fallot with pulmonary regurgitation. PMID- 9874074 TI - Another common noun derived from a proper name. PMID- 9874075 TI - A case of aortocoronary dissection as a complication during a percutaneous transluminal coronary angioplasty (PTCA). AB - We report a case of aortocoronary dissection during a percutaneous transluminal coronary angioplasty (PTCA) which resulted from an antegrade and also a progressively retrograde extension of the coronary dissection into the Sinus of Valsalva and the ascending aorta. It was successfully treated with stenting without an operation, resulting in optimal coronary blood flow and diminution of the aortic dissection. PMID- 9874076 TI - Cardiorespiratory response during exercise in patients with cyanotic congenital heart disease with and without a Fontan operation and in patients with congestive heart failure. AB - To clarify the different cardiorespiratory response to exercise in patients with congenital heart disease and patients with chronic congestive heart failure, we investigated the effect of a progressive exercise test in 30 patients aged 10 to 24 years, including 9 patients with cyanotic congenital heart disease (group A), 13 patients who had undergone a Fontan operation (group B), and 8 patients with reduced left ventricular function (group C), and 18 healthy controls (group D). There was no difference in peak oxygen uptake among patient groups and all group A, B, and C values were lower than those in group D (P<0.001). Although peak heart rate was lower in patient groups than in group D, heart rate at a given exercise intensity was highest in group C. The oxygen pulse (oxygen uptake divided by heart rate=stroke volume x arterial venous oxygen difference), as an indicator of stroke volume, was lower in patients groups, especially in group C, than in group D. There was no difference in tidal volume between groups A and D, but the respiratory rates at any given exercise intensity were higher in group A than in the other patient groups, thus minute ventilation and the ventilatory equivalent were highest in group A. The increased respiratory rate and low tidal volume in group C resulted in rapid and shallow respiration. There was no difference in exertional symptoms at peak exercise among the groups. In addition to impaired responses of stroke volume during exercise in patients with reduced exercise capacity, there was little limitation of increase in ventilation in group B and excessive ventilation in group A. The present results suggest that relationship between ventilatory and cardiac responses during exercise in patients with cyanotic congenital heart disease with and without a Fontan operation is different from the relationship in patients with chronic congestive heart failure; however, these pathological differences did not influence exertional symptoms. PMID- 9874077 TI - Parasympathetic withdrawal antedates dynamic myocardial ischemia in patients with syndrome X. AB - This study was to evaluate the dynamic changes in cardiac autonomic control preceding electrocardiographic (ECG) myocardial ischemia in patients with syndrome X. Twenty-four-hour ambulatory ECG was obtained in 34 consecutive patients in a drug-free state. Fourteen (41%) of them, aged 58.8+/-13.5 years, presented a total of 19 ischemic episodes, mean duration 12.4+/-19.8 min (ranged 1 to 90 min). Heart rate variability was measured for 24 h; for 3 min and 30 min before, and during the 15 min (in five 3-min intervals) immediately antedating ST segment depression; and for another 3 min after ST segment back to normal. There were significant progressive shortenings in sinus cycle lengths over the 30 min preceding myocardial ischemia (-30 vs -3 minute, 822+/-32 ms vs 637+/-23 ins, P<0.05; a decrement of 22.5%). The sinus cycle lengths lengthened after ischemia ceased. High frequency activity, pNNSO and rMSS.D. were significantly reduced from the -30 min baseline to a nidus in the last 3 min before ischemia (P<0.05), whereas low frequency band and low/high frequency ratio did not present significant change. These findings strongly argue that cardiac autonomic control, especially vagal withdrawal, is involved in the pathogenesis of dynamic myocardial ischemia in syndrome X. PMID- 9874078 TI - Clinical experience with downsized lower energy output implantable cardioverter defibrillators. Ventak Mini II Clinical Investigators. AB - BACKGROUND AND STUDY OBJECTIVE: Technical improvements in cardioverter defibrillators technology has resulted in decrease in can size coupled with improved electrodes technology. A decrease in maximum energy output allows further decrease in device size. The aim of this study was to evaluate the feasibility of a single lead transvenous implant employing a downsized cardioverter-defibrillator (volume 59 cm3), with a related decrease in maximum energy output (29-31 joules as stored energy and 25-27 joules as delivered energy). METHODS AND RESULTS: Fifty-five patients with ventricular tachyarrhythmias were enrolled in 17 European institutions for implantation. At implantation step-down defibrillation threshold (DFT) was determined and the device was implanted only if a safety margin > or =10 joules was maintained between DFT and maximum programmable output. Implantation was performed in 54 of the 55 referred patients (98%) in a single electrode-device configuration. Step down DFT testing was performed in 44 patients (43 finally implanted) and DFT was 7.77+/-4.41 joules (range 3-20). In 20 of the tested patients (45%) DFT was < or =5 joules, in 26 patients (59%) was < or =8 joules and in 34 patients (77%) it was < or =10 joules. No differences were found in DFT comparing patients with left ventricular ejection fraction < or = or >40% or patients treated or not with antiarrhythmic drugs or beta-blockers. Mean implant duration was 85+/-34 min. CONCLUSIONS: Employing a downsized cardioverter defibrillator, successful transvenous implantation can be achieved in 98% of the patients, with maintenance of adequate defibrillation safety margins despite a reduction in stored energy to 29 joules. PMID- 9874079 TI - Association of increased QT dispersion with coronary atherosclerosis in patients with aortic stenosis. AB - To evaluate correlates between electrocardiographic QT dispersion and coronary atherosclerosis in patients with aortic stenosis before aortic valve replacement, 39 consecutive patients >40 years old with symptomatic aortic stenosis and coronary diameter narrowing > or =50% measured by digital angiographic study were included. An additional matched group with insignificant coronary lesions (<50%) consisted of 39 patients for comparisons. Matching by age, sex heart rate and incidence of chest pain resulted in two comparable groups with identical baseline characteristics. Preoperative transthoracic echocardiography and electrocardiograms were performed in all subjects. QT dispersion was defined as the difference between maximal and minimal QT interval measurements occurring among any of the 12 leads on a standard electrocardiogram. No subject had fewer than nine measurable leads. There were no significant differences of risk factors of coronary artery disease between the two groups. From a conditional multivariate logistic regression analysis, independent predictors of development of coronary artery disease in aortic stenosis were only QTc dispersion (odds ratio= 1.255, P=0.01). A wide QTc dispersion > or =70 ins) correlated with the presence of angiographically significant coronary artery disease with a sensitivity and specificity of 72% and 79%. The positive accuracy of having significant coronary artery disease in the presence of QTc dispersion > or =70 ms was 78%. The negative predictive value was 74%. In conclusion, electrocardiographic QTc dispersion may provide important clinical information. A wide QTc dispersion in patients with aortic stenosis is associated with a high incidence of coronary artery disease. These findings warrant further investigation in a large trial. PMID- 9874080 TI - Changes of autonomic tone before the onset of paroxysmal atrial fibrillation. AB - The relationship between autonomic nerve system and the onset of paroxysmal atrial fibrillation (PAF) is still controversial. Furthermore, no prior studies have compared heart rate variability (HRV) between PAF patients with (organic) or without (idiopathic) underlying cardiac diseases. The purpose of this study was to assess the alteration of autonomic tone by analyzing HRV immediately before the onset of atrial fibrillation. This study included 57 patients (M/F: 34/23, 66+/-22 years) with frequent attacks of PAF. All cases underwent 24-h ambulatory Holter monitoring; each patient had one or more episodes of sustained PAF (>30 s). A period of sinus rhythm 40 min was allowed for accurate assessment of HRV over these periods. Spectral HRV was expressed as low (0.04-0.15 Hz) and high (0.15-0.40 Hz) frequency components (LF, HF), and L/H ratio at 2-min intervals over a 40-min period before the onset of PAF. According to HRV, three subtypes were classified; onset of PAF accompanied with increased HF component and decreased L/H ratio was designated as vagal type; decreased HF component and increased L/H ratio was designated as sympathetic type, and the other episodes which did not belong to vagal or sympathetic type were designated as non-related type. In group I (idiopathic PAF, n=30): 63 episodes of PAF were found and vagal type was predominant (41/63, 63.5%); HF increased significantly before the onset of AF. In group II (organic PAF, n=27): 58 episodes of PAF were found and sympathetic type was predominant (39/58, 67.2%); L/H ratio increased before AF onset. None of the three subtypes showed significant circadian distributions in group I and II patients. Changes of HRV before the onset of PAF were not universal; most of the patients with idiopathic PAF were vagal dependent and most of the patients with organic PAF were sympathetic dependent. PMID- 9874081 TI - Evolution of surgical trends in congenital heart disease: a population based study. AB - The aim of this study was to establish surgical trends in patients with congenital heart disease operated on between 1947 and 1997 in a population based study. All patients diagnosed as having congenital heart disease, born in Malta up to 1995 inclusive and operated for congenital heart disease up to 1997 inclusive were included. Analysis was carried out for lesions operated, age at surgery, operative centre and mortality rates, in the setting of a regional hospital providing congenital heart disease diagnostic and follow-up services for all Malta. Increasingly more operations for cardiac malformations are being carried out, with a progressively higher proportion of operations performed on complex conditions (P<0.001), at an ever younger age (P<0.001), and with a declining perioperative mortality (P<0.001). For the period 1990-1994, 4.2 operations for congenital heart disease/1000 live births were required. Factors which may increase or decrease this rate in future are discussed, along with costs of surgery. Surgery for congenital heart disease has become progressively more aggressive and safer since this method of treatment for these malformations was initiated, but this has occurred at a significant financial cost. PMID- 9874082 TI - Diastolic function and insulin resistance in essential hypertension. AB - We studied the relationship of insulin metabolism to diastolic function in 41 hypertensive patients and 24 healthy subjects. They underwent a 75-g oral glucose tolerance test. Integrated response of insulin to the glucose load was calculated as the area under the curve of insulin (AUC-I) and glucose (AUC-G). The ratio AUC I/AUC-G was taken as an index of insulin resistance. Echocardiographic examination was performed to measure left ventricular mass. We used pulsed Doppler technique to assess the ratio of early to late transmitral peak velocity (E/A). The hypertensives had higher fasting insulin concentrations, insulin levels at 60 min and at 120 min after oral glucose load. They had higher AUC-I and higher index of insulin resistance compared to normotensives. Patients with hypertension had a lower E/A ratio in comparison to normotensive volunteers. In all subjects, the E/A ratio was negatively correlated with fasting insulin and insulin concentrations at 30, at 60 and at 120 min, AUC-I and insulin resistance index. Multiple regression analysis was performed to evaluate the above relationships, when the effects of confounding factors such as age, heart rate and systolic blood pressure were taken into account. We found that E/A ratio is negatively correlated to insulin concentrations at 120 min and to AUC-I in hyperinsulinemic subgroup of subjects (fasting insulin> 10). We conclude, that insulin metabolism and E/A ratio, which reflects diastolic function of the left ventricle, are related in hypertension. PMID- 9874083 TI - Quantification of mitral and tricuspid regurgitation by the proximal flow convergence method using two-dimensional colour Doppler and colour Doppler M mode: influence of the mechanism of regurgitation. AB - In patients with mitral (n=77: organic=49, functional=28) and tricuspid regurgitation (n=55: functional=54) quantified by angiography, the temporal variation of the proximal flow convergence region throughout systole was assessed by colour Doppler M-Mode, and peak and mean radius of the proximal isovelocity surface area for 28 cm/s blood flow velocity were measured. Additionally, the peak radius derived from two-dimensional colour Doppler was obtained. About 50% of the patients with mitral and tricuspid regurgitation showed a typical temporal variation of the flow convergence region related to the mechanism of regurgitation. The different proximal isovelocity surface area radii were similarly correlated to the angiographic grade in mitral and tricuspid regurgitation (rank correlation coefficients 0.55-0.89) and they differentiated mild to moderate (grade < or =II) from severe (grade > or =III) mitral and tricuspid regurgitation with comparable accuracy (82-96%). However, moderate mitral regurgitation due to leaflet prolapse in two patients was correctly classified by the mean M-mode radius and overestimated by both peak radii. Only half of the patients showed a typical variation of the flow convergence region related to the mechanism of regurgitation. The different proximal isovelocity surface area radii were suitable to quantify mitral and tricuspid regurgitation in most patients. However, in mitral regurgitation due to leaflet prolapse the use of the mean M-mode radius may avoid overestimation. PMID- 9874084 TI - Double chambered right ventricle: delineation by multiplane transoesophageal echocardiography. AB - Three consecutive patients (median age 5 years; median weight 15 kg) with double chambered right ventricle (DCRV) were studied by multiplane transoesophageal echocardiography (TOE), using a specially designed paediatric probe. Using a 30 degrees angulation from the longitudinal plane, the body and outflow tract of the right ventricle could be imaged with minimal foreshortening, allowing detailed description of the level and nature of obstruction. In all cases, angiography and subsequent surgical inspection confirmed the TOE findings. Multiplane TOE should avoid the necessity for angiography in the preoperative assessment of this unusual lesion. PMID- 9874085 TI - Separate origins of left anterior descending, left circumflex and intermediate arteries from the left coronary aortic sinus. AB - We describe a patient with an unusual coronary anatomic variance in which separate origins of the left anterior descending, left circumflex and the intermediate arteries arose from the left coronary aortic sinus. The importance of demonstrating the separate ostium of an intermediate artery is emphasized. PMID- 9874086 TI - Bone marrow suppression and klebsiella pneumonia septicemia due to ticlopidine and successful treatment with filgrastim; a case report. AB - In this report we describe an elderly lady, who after 27 days of ticlopidine treatment developed severe pancytopenia and gram (-) septicemia. No clinical response was obtained with an eight days course of empirical broad spectrum antibiotics, after which granulocyte colony stimulating factor (G-CSF, filgrastim) added to her regimen. In a period of seven days her neutrophil count was normal and she had recovered from septicemia. Twelve weeks after discharge she is fine and her blood parameters are all normal. PMID- 9874087 TI - Successful management of right coronary air embolus after intracoronary administration of verapamil. AB - A case of a 50 year-old male with a coronary air embolism is described. The case was successfully treated with intracoronary administration of verapamil. PMID- 9874088 TI - Is atherosclerosis an overreaction of the immune system? PMID- 9874089 TI - Reciprocal dopamine-glutamate modulation of release in the basal ganglia. AB - Dopaminergic and glutamatergic transmissions have long been known to interact at multiple levels in the basal ganglia to modulate motor and cognitive functions. One important aspect of their interactions is represented by the reciprocal modulation of release. This topic has been the object of interest since the late 70's, particularly in the striatum and in midbrain dopaminergic areas (substantia nigra and ventral tegmental area). Analysis of glutamate-dopamine interactions in the control of each other's release is complicated by the fact that both glutamate and dopamine act on multiple receptor subtypes which can exert different effects. Therefore, glutamatergic modulation of dopamine release has been reviewed by analyzing the effects of glutamatergic selective receptor agonists and antagonists in the striatum (both motor and limbic portions) and in midbrain dopaminergic areas, as revealed by in vitro (slices, cell cultures, synaptosomes) and in vivo (push-pull, microdialysis and voltammetry techniques) experimental approaches. The same approach has been followed for dopaminergic modulation of glutamate release. The facilitatory nature of glutamate modulating both presynaptic and dendritic dopamine release has clearly emerged from in vitro studies. However, evidence is presented that, at least in the striatum and in the nucleus accumbens of awake rats, glutamate-mediated inhibitory effects may also occur. In vitro and in vivo experiments in the striatum and midbrain dopaminergic areas mainly depict dopamine as an inhibitory modulator of glutamate release. However, in vivo studies reporting dopamine D1 receptor mediated facilitatory effects are also considered. Therefore, the general notion that glutamate and dopamine act oppositely to regulate each other's release, is only partly supported by the available data. Conversely, the nature of the interaction between the two neurotransmitters seems to vary depending on the experimental approach, the brain area considered and the subtype of receptor involved. PMID- 9874090 TI - Matching kinetics of synaptic vesicle recycling and enhanced neurotransmitter influx by Ca2+ in brain plasma membrane vesicles. AB - Using native plasma membrane vesicle suspensions from the rat cerebral cortex under conditions designed to alter intravesicular [Ca2+], we found that Ca2+ induced 47 +/- 5% more influx of [3H]GABA, [3H]D-aspartate and [3H]glycine at 37 degrees C with half-times 1.7 +/- 0.5, 1.3 +/- 0.4 and 1.3 +/- 0.4 min, respectively. We labelled GABA transporter sites with the uptake inhibitor, [3H] (R,S)-N-[4,4-bis(3-methyl-2-thienyl)but-3-en-1-yl]nipecotic acid and found that Ca2+ induced a partial dissociation of the bound inhibitor from GABA transporter sites with a similar half-time. By means of rapid kinetic techniques applied to native plasma membrane vesicle suspensions, containing synaptic vesicles stained with the amphipathic fluorescent styryl membrane probe N-(3 triethylammoniumpropyl)-4-[4-(dibutylamino)styryl]pyrid inium dibromide, we have measured the progress of the release and reuptake of synaptic vesicles in response to Ca2+ and high-[K+] depolarization in the 0.0004-100 s range of time. Synaptic vesicle exocytosis, strongly influenced by external [Ca2+], appeared with the kinetics accelerated by depolarization. These results are consistent with the potential involvement of Ca2+ in taking low-affinity transporters to the plasma membrane surface via exocytosis. PMID- 9874091 TI - Propionic and methylmalonic acids inhibit the in vitro phosphorylation of a 85 kDa cytoskeletal protein from cerebral cortex of rats. AB - In this study we examine the action of methylmalonic (MMA) and propionic (PA) acids, metabolites which accumulate in methylmalonic and propionic acidemias respectively, on the endogenous phosphorylating system associated with the cytoskeletal fraction of cerebral cortex of young rats. Chronic treatment with PA and treatment of tissue slices with MMA or PA are effective in decreasing the in vitro phosphorylation into a 85 kDa cytoskeletal associated protein. We tested the effect of the acids on the endogenous kinase activities by using specific kinase activators and inhibitors. Results demonstrated that the acids interfere with the endogenous cAMP-dependent and Ca2+/calmodulin-dependent kinase activities. Furthermore, in vitro dephosphorylation of the 85 kDa protein was totally inhibited in brain slices treated with the acids. Considering the importance of protein phosphorylation to cellular function, we speculate that alteration in the phosphorylating level of cytoskeletal associated phosphoproteins induced by MMA and PA treatments may somehow be involved in steps leading to brain damage. PMID- 9874092 TI - Semicarbazide-sensitive amine oxidase (SSAO) from human and bovine cerebrovascular tissues: biochemical and immunohistological characterization. AB - Semicarbazide-sensitive amine oxidase (SSAO) is widely distributed in almost all tissues, especially in vascularized ones. However, its presence in brain microvessels is still controversial. We have investigated the presence of SSAO in human and bovine brain microvessels by biochemical and immunohistological techniques, and we have compared it with SSAO present in meninges from the same species. SSAO metabolizes benzylamine and methylamine in all tissues tested and possibly dopamine and octopamine as well, as shown in competition studies. Kynuramine inhibited the metabolism of benzylamine by SSAO with high affinity in a non-competitive manner. Western-blot analysis rendered a positive staining of a 100 kDa band, in tissues from both species. These results were confirmed by immunohistological studies: the tunica media and intima of the meninges from both species were positively stained, and so was the endothelial layer of microvessels. SSAO was absent in brain parenchyma. These results definitively confirm the presence of SSAO in human and bovine cerebrovascular tissues and they demonstrate for the first time, the presence of this amine oxidase in endothelial cells from microvessels, through biochemical and immunological approaches. PMID- 9874093 TI - Na+,K+-ATPase interaction with a brain endogenous inhibitor (endobain E). AB - Na+,K+-ATPase activity of rat brain synaptosomal membranes was evaluated in the presence of an inhibitory fraction II-E (termed endobain E), isolated by gel filtration and anionic exchange HPLC of a rat brain soluble fraction. We studied endobain E aging, analyzed its inhibitory potency in the absence or presence of ouabain as well as its ability to block high affinity [3H]ouabain binding to cerebral cortex membranes. Similar loss of endobain E activity was observed when samples were stored either dried or in solution. Endobain E fraction inhibited synaptosomal membrane Na+,K+-ATPase activity in a concentration-dependent manner and the slope of the corresponding curve strongly resembled that of ouabain. Assays performed in the presence of endobain E and ouabain indicated that the inhibitory effect was additive or less than additive, depending on their respective concentrations during preincubation and/or incubation. High affinity [3H]ouabain binding to cerebral cortex membranes proved concentration-dependent from 0.10 to 0.50 mg protein per ml; binding inhibition by endobain E was independent of protein concentration within the above range. [3H]ouabain binding inhibition by endobain E was concentration-dependent over a 10-fold range, an effect similar to that found for Na+,K+-ATPase inhibition. The extent of endobain E effect on Na+,K+-ATPase inhibition was much higher (90-100%) than that on [3H]ouabain binding blockade (50%). Findings suggest some type of interaction between endobain E and ouabain inhibitory mechanisms and favour the view that the former behaves as an endogenous ouabain. PMID- 9874094 TI - Acute effects of acetyl-L-carnitine on sodium cyanide-induced behavioral and biochemical deficits. AB - In the present study we investigated the effects of acute treatment with acetyl-L carnitine (50 mg/kg, i.v. 90 min before the sodium cyanide injection) on a sodium cyanide-induced behavioral deficit in the Morris water escape task. In a first experiment the spatial discrimination performance of the rats was found to be dose-dependently impaired after an i.c.v. injection of sodium cyanide (2.5 and 5.0 microg). Acute treatment with acetyl-L-carnitine was found to increase the behavioral deficit after sodium cyanide. These findings were replicated in a second experiment. Based on these results it can be argued that an acute administration of acetyl-L-carnitine appears to potentiate a sodium cyanide induced behavioral deficit. An additional in vitro experiment with rat brain synaptosomes showed clear effects of administered sodium cyanide on the energy dependent incorporation of inositol into phosphoinositides and on the ATP concentration. In vitro acetyl-L-carnitine administration had no effect on the sodium cyanide-induced energy depletion. The negative behavioral findings are in contrast with our previously found protective effect of chronic treatment with acetyl-L-carnitine (via drinking water) on the sodium cyanide-induced behavioral deficit. Since chronic acetyl-L-carnitine treatment has no effect on the phosphoinositide metabolism it was suggested that acetyl-L-carnitine may act via the formation of an ATP-independent reservoir of activated acyl groups. Thus, fatty acids as acylated derivatives can be used for reacylation processes during an acute period of energy depletion. However, we have no clear explanation for the discrepancy in behavioral results between the chronic vs acute treatment of acetyl-L-carnitine at present. Further research is needed to characterize the mechanism of action of acetyl-L-carnitine in relation to sodium cyanide. PMID- 9874095 TI - Receptor systems participating in nicotine-specific effects. AB - It is generally accepted that self-administration of drugs is prompted primarily by a reward system driven by an increase in extracellular dopamine in the nucleus accumbens. Recent findings that dopamine increase in the accumbens can be caused by many other factors, among them stress, suggest a more complex mechanism, and possibly differences in the reward system for different compounds. In the present paper we compare the effects of receptor-specific antagonists on the increase of dopamine induced by nicotine with that induced by cocaine in the nucleus accumbens in conscious rats. The compounds alone or together were injected intravenously, and dopamine level changes were measured via microdialysis. When administered together the effect of nicotine and cocaine on the level of dopamine in the accumbens was additive. Apparently there is some interaction between the two compounds, since nicotine had no effect after combined nicotine and cocaine administration. Perhaps the available dopamine pool was exhausted by the prior administration. The nicotinic antagonist mecamylamine, the muscarinic antagonist atropine, and the NMDA glutamate receptor antagonist MK-801 each blocked nicotine induced dopamine release in the accumbens, indicating the participation of more than a single receptor system in the nicotine-induced effect. These three antagonists did not inhibit cocaine-induced dopamine increase in the accumbens, indicating the lack of a role of these receptors in the cocaine effect under our experimental conditions. SCH-23390, a dopamine D1 receptor antagonist, blocked both nicotine- and cocaine-induced effects, indicating the possible role of this receptor in these reward effects. The results indicate that there are differences in some of the receptors mediating the central effects of the two compounds examined, nicotine and cocaine, although each influences dopamine levels, and that the two compounds interact. PMID- 9874096 TI - Characterization of both dopamine uptake systems in rat striatal slices by specific pharmacological tools. AB - Previous results have shown that modifications of dopamine (DA) high-affinity uptake1 and those of DA low-affinity uptake2 in rat striatal slices were different after autoxidation of this model and in the presence of antioxidants. The aim of this study was to determine whether these two DA uptake systems correspond to two different dopamine transporters or rather to a single one. A lesion into the substantia nigra of animals by injection of 6-hydroxydopamine, a neurotoxic substance of nigrostriatal dopaminergic neurons led to the suppression of both DA uptake systems. These two DA uptake systems were not modified when animals were treated by reserpine or tetrabenazine, which inhibit the vesicular monoamine transporter. Moreover, they were sodium- and temperature-dependent. Experiments with specific inhibitors showed that 1-[2-(diphenylmethoxy) ethyl]-4 (3-phenylpropyl)-piperazine dihydrochloride (GBR-12935) and (E)-N-(3-iodoprop-2 enyl)-2beta-carbomethoxy-3beta-(4'-tolyl ) nortropane chloride (PE2I), two selective DA uptake inhibitors, were significantly more potent than fluoxetine and nisoxetine (selective serotonin and norepinephrine uptake inhibitors respectively) in both DA uptake systems. However, the concentrations of these products inhibiting low-affinity uptake2 by 50% were much greater than those for high-affinity uptake1. Our data indicate that both DA uptake systems are neuronal, independent of the vesicular monoamine transporter, active and specific for dopamine. Our results suggest that high-affinity uptake1 and low-affinity uptake2 correspond to the same dopamine transporter, but would be situated at different levels in the striatal slice model. Uptake1 could take place at the periphery of the slice whereas uptake2 in the depth of the slice. PMID- 9874097 TI - The role of taurine in the regulation of brain cell volume in chronically hyponatraemic rats. AB - Cells in slices prepared from the superficial cerebral cortex of normonatraemic rats underwent moderate swelling when exposed to low Na+ medium (122 mmol/l) accompanied by a large increase in the rate of efflux of preloaded taurine. In contrast, cells in slices from chronically (4 day) hyponatraemic rats did not increase in volume and the rate of taurine efflux was unchanged. The anion transport inhibitor 4,4'-diisothiocyanato-stilbene-2,2'-sulphonic acid (25 micromol/l) caused marked (-44%) reduction in taurine efflux in cells from normonatraemic rats; this response was strongly attenuated (-16%) by hyponatraemia. When slices from hyponatraemic rats were acutely exposed to medium containing 142 mmol/Na+ cells exhibited marked and paradoxical swelling. This response was completely abolished by the NaCl co-transport inhibitor bumetanide (50 micromol/l) and was not observed in slices that had not been pre-loaded with taurine. Forty eight hours after the start of the remission of hyponatraemia, cells from post-hyponatraemic rats displayed normal responses (i.e., moderate swelling and greatly accelerated taurine efflux) on exposure to 122 mmol/Na+. But at 24 h there was only partial restoration of the efflux response to 122 mmol/Na+, with an enhanced cell swelling response that was not significantly affected by bumetanide. It is concluded that (i) during chronic hyponatraemia, unlike acute hyposmotic stress, cortical cells preserve their volume and that this is not associated with any increase in the rate of taurine loss; there does however, appear to be a decrease in the anionic component of cellular taurine efflux; (ii) acute re-incubation of slices in medium containing 142 mmol/l Na+ is associated with cell swelling that may reflect up-regulation of Na/Cl/taurine co transport; (iii) following restoration of normonatraemia the pattern of normal cellular response to acute hyposmotic stress is only gradually re-established. PMID- 9874098 TI - Survey of seroprevalence of Q fever in dogs in the southeast of France, French Guyana, Martinique, Senegal and the Ivory Coast. AB - A serological survey was carried out on 429 dogs belonging to the French military in France, French Guyana, Martinique, Senegal and the Ivory Coast. Serology against phase I and II antigens of Coxiella burnetii, the intracellular zoonotic bacterium was performed using indirect immunofluorescence techniques. Specific antibodies were found in dogs from France (9.8%), Senegal (11.6%), Ivory Coast (8.3%), French Guyana (5.2%) but not in those from Martinique. The seroprevalence among 77 dogs who had contact with sheep compared with 352 dogs who had had no contact, demonstrated a significantly higher seroprevalence in the former. Our results indicate that dogs, living close to sheep, may be infected by Coxiella burnetii and should be considered as possible sources of infection for humans. PMID- 9874099 TI - Sensitive detection and typing of porcine reproductive and respiratory syndrome virus by RT-PCR amplification of whole viral genes. AB - Following the recent use of a live vaccine against porcine reproductive and respiratory syndrome virus (PRRSV) in Denmark, both American (vaccine) and European-type PRRSV now coexist in Danish herds. This situation highlighted a requirement for supplementary tests for precise virus-typing. As a result, we developed a RT-PCR assay able to detect as well as type PRRSV. To provide maximal sequence information, complete viral open reading frames (ORFs 5 and 7) were targeted for amplification. The RT-PCR test was able to amplify complete PRRSV ORFs from complex materials such as boar semen containing as little as 1 TCID50 ml(-1) of PRRSV. Typing of viruses was accomplished by any one of three strategies: (i) use of type-specific PCR primers, (ii) size determination of ORF 7 amplicons, (iii) DNA sequencing. All three typing strategies showed complete concordance with the currently used method of typing with monoclonal antibodies (MAbs) when used on a panel of PRRSV field isolates covering the period 1992 1997. The ORF 7-based test had particularly desirable characteristics, namely, highly sensitive detection of PRRSV without apparent type bias, typing of the detected virus, discrimination between pure and mixed virus populations, and semi quantitative assessment of type ratios in mixed populations, all in a single PCR reaction. In addition, the obtained sequence data were used to predict two simple and rapid strategies (single-enzyme restriction length polymorphy analysis and oligonucleotide hybridization) for confirmation of the specificity of ORF 7 RT PCR reactions. As such, the RT-PCR assay provides a new, powerful diagnostic tool to study the population dynamics between present and emerging PRRSV-types. PMID- 9874100 TI - Distribution of bovine virus diarrhoea virus in tissues and white blood cells of cattle during acute infection. AB - This study is performed to gain knowledge about the quantitative distribution of bovine virus diarrhoea virus (BVDV) in tissues and white blood cells (WBC) at different intervals after acute infection. Ten specific pathogen-free calves were intranasally inoculated with 10(5) 50% tissue culture infective dose of the non cytopathic BVDV strain 4800. Twelve hours after inoculation tonsil biopsies were taken and WBC were collected daily for virus isolation and titration. Each day one calf was killed and virus isolations and titrations were performed from a range of tissues. The results indicate that BVDV first replicates in nasal mucosa and to high titers in the tonsil. The virus then appeared to spread to the regional lymph nodes and then disseminates throughout the body. The virus titers were highest in tonsil, thymus and ileum and were low in the WBC. Also after in vitro infection virus titers in WBC were very low, whereas, they were high in epithelial cells. Although the WBC might not be as important as other cells for replication of BVDV, they may play a role in the spread of the virus throughout the body. PMID- 9874101 TI - Observations on thermostable subpopulations of the unconventional agents that cause transmissible degenerative encephalopathies. AB - When scrapie agent is exposed to partially inactivating autoclave cycles, the fraction of infectivity that survives remains thermostable during relatively long periods of autoclaving. This resistant subpopulation can also be differentiated from the main population by its prolonged incubation periods in assay animals, compared with control material. Stabilisation of this subpopulation may occur through the smearing and drying of infected tissue that can occur prior to autoclaving, in which the disease-specific form of PrP protein (PrP(Sc)) could become rapidly heat-fixed. This may paradoxically be what protects this fraction of PrP(Sc) from further inactivation during autoclaving. Data are presented showing that the thermostability acquired by the resistant subpopulation is a stable characteristic; autoclaving for a second time results in very little further loss of infectivity. These observations suggest that inactivation procedures that do not involve rapid and effective fixation of PrP(Sc) may be better candidates for dealing effectively with scrapie-like agents. PMID- 9874102 TI - The specificity of antibody in chickens immunised to reduce intestinal colonisation with Campylobacter jejuni. AB - Poultry consumption has been identified as a major risk factor for human infection with Campylobacter jejuni in developed countries. C. jejuni is present in the gastrointestinal tract of broiler chickens at the time of slaughter, and faecal contamination of carcases during processing results in significant campylobacter loads on carcases. One approach to reducing the level of carcase contamination with C. jejuni is to control campylobacter infection in broiler chickens. To this end, the study described here investigated the specificity of antibody in serum and intestinal secretions of chickens that had been immunised with campylobacter antigens and then challenged with viable bacteria. The immunodominant antigens in the serum of birds that showed a 2-log reduction in caecal colonisation with C. jejuni included flagellin protein (61-63 Kd) and three additional antigens of 67, 73.5 and 77.5 Kd. Only flagellin and the 67 Kd antigen were recognised by IgG antibody in gastrointestinal secretions of the same birds. Antibody from chickens immunised with purified native flagellin protein recognised flagellin protein and the 67 Kd antigen in Western blots probed with serum, but only the flagellin proteins (61-63 Kd) in Westerns probed with gastrointestinal secretions. Analysis of the specificity of the response to flagellin protein using recombinant clones that expressed regions of the flagellin gene suggests that epitopes in each region of the flagellin protein were immunogenic. Of the immunodominant antigens, only flagellin appeared to be surface-exposed on viable C. jejuni, although conformational epitopes of flagellin appeared to be sensitive to the method of antigen purification. The results of this study suggest that flagellin and possibly the 67 Kd antigen may be valuable for immunological control of intestinal infection with C. jejuni in chickens, but that further work is required to purify these as vaccine candidates by using methods that preserve conformational epitopes. PMID- 9874103 TI - A method for purification and characterisation of Mycobacterium avium subsp. paratuberculosis from the intestinal mucosa of sheep with Johne's disease. AB - Mycobacterium avium subsp. paratuberculosis, the cause of Johne's disease in ruminants, cannot be cultured in large quantities from affected sheep in Australia. A method is described for the purification of the organism from the intestinal mucosa of sheep with multibacillary Johne's disease in order to undertake restriction fragment length polymorphism (RFLP) analysis for epidemiological purposes. Using sucrose and potassium chloride as separation media for differential and density gradient centrifugation, yields of approximately 90 mg dry weight M. avium subsp. paratuberculosis per 5 g intestinal mucosa were obtained. The preparations of purified M. avium subsp. paratuberculosis were visually free of non-acid fast bacteria and contained 10(2) 10(3) aerobic/ facultatively anaerobic organisms per gram wet weight. DNA extracted from purified M. avium subsp. paratuberculosis was examined by hybridisation with an IS900 probe after digestion with BstEII and RFLP patterns distinct from isolates from cattle were obtained. The RFLP pattern of purified M. avium subsp. paratuberculosis from five sheep matched that obtained previously from organisms cultured from sheep in studies in New Zealand, indicating that the purification and RFLP method is robust. PMID- 9874104 TI - Efficacy of ELISA compared to conventional tests (RBPT and CFT) for the diagnosis of Brucella melitensis infection in sheep. AB - An indirect ELISA (iELISA) using a B. abortus smooth lipopolysaccharide (S-LPS) as coating antigen and a polyclonal anti-sheep IgG peroxydase-labeled serum as conjugate was developed. The iELISA was assessed in comparison to the Food and Agriculture Organisation/International Atomic Energy Agency (FAO/IAEA) standard bovine ELISA kit (IAEA kit) using an anti-bovine IgG conjugate that cross-reacts with sheep antibodies, and with complement fixation test (CFT) and Rose Bengal Plate Test (RBPT). The evaluation was performed on sera from ewes experimentally infected with Brucella melitensis 53H38 (H38), using negative and positive sheep reference sera. No significant difference was found as regards to the specificity, the lower limit of detection and the estimated sensitivity of the two iELISAs. This suggests that an anti-bovine conjugate could allow the development of only one ELISA protocol for all ruminants. The iELISA, if well standardized, proved to be a good screening test able to be used either alone or in addition to RBPT. PMID- 9874106 TI - Molecular variants of HIV-1 and their impact on vaccine development. PMID- 9874105 TI - A study of relationships among F17 a producing enterotoxigenic and non enterotoxigenic Escherichia coli strains isolated from diarrheic calves. AB - We investigated the clonal relationships among 41 enterotoxigenic (ETEC) or non enterotoxigenic (NETEC) Escherichia coli strains producing the F17 a fimbriae isolated from diarrheic calves in France or Belgium in the early 1980s. Twenty three of the 26 ETEC strains were highly clonally related, most of them with a O101:K32:H9-serotype. The NETEC strains were also divided in clonal subgroups, most of them with O101:H-serotype. The F17 a positive ETEC strains are no longer isolated from diarrheic calves in these countries. It is postulated that the use of a vaccine including O101, K32 and H9 antigens in addition to K99 (F5) explains the strongly reduced isolation of the O101:K32:H9, K99 (F5) E. coli clone. PMID- 9874107 TI - Towards an AIDS vaccine: the role of primate models. PMID- 9874108 TI - Strategies for the prevention and treatment of sexually transmitted diseases. PMID- 9874109 TI - Effectiveness, impact and cost of syndromic management of sexually transmitted diseases in Tanzania. PMID- 9874110 TI - STD surveillance in the Russian Federation. PMID- 9874111 TI - Women and AIDS: a brief update. PMID- 9874112 TI - Reduction of mother-to-child transmission of HIV infection: non-pharmaceutical interventions and their implementation. PMID- 9874113 TI - Perinatal transmission--successful interventions. Where do we go from here? PMID- 9874114 TI - The role of proteinases in the treatment of HIV infection. PMID- 9874115 TI - Future trends in Kaposi's sarcoma and lymphoma. PMID- 9874116 TI - Preventive HIV-1 vaccines: where are we going? PMID- 9874117 TI - The management of adult male victims of sexual assault in the GUM clinic: a practical guide. PMID- 9874118 TI - An outbreak of methicillin-resistant Staphylococcus aureus (MRSA) infection in HIV-seropositive persons. AB - Staphylococcus aureus is a cause of considerable morbidity and mortality in HIV seropositive persons. Although methicillin-resistant S. aureus (MRSA) is encountered worldwide and in many areas of medical care, little has been reported on clinical infection with MRSA in patients with HIV. We report on an outbreak of MRSA infection in HIV antibody positive patients, using case reports to describe an outbreak of MRSA infection in HIV-seropositive persons. Six cases of clinical MRSA infection were reported over a 4-week period on patients on an HIV dedicated ward. All cases had previous AIDS diagnoses and low CD4 cell counts (median 8 x 10(6)/l; range 0 to 238). Two cases had infected skin lesions and 2 cases had infected indwelling central venous catheters with septicaemia. Two cases had pneumonia, one with concurrent infection at the entry site of a percutaneous endoscopic gastrostomy (PEG) feeding tube. Isolates of MRSA from the 6 cases were compared by pulsed-field gel electrophoresis of Sma1 chromosomal digests. The resultant banding pattern showed the same strain was responsible for all the infections. A seventh inpatient, the index case, had positive carriage with the same strain of MRSA. To define ongoing MRSA carriage after the outbreak, 29 consecutive ward patients were swabbed for MRSA: all were negative. All patients identified with MRSA infection responded to treatment with intravenous teicoplanin, although carriage was unaltered. Four of the 6 cases died within 7 weeks of diagnosis of MRSA. MRSA can cause severe morbidity in patients with end stage HIV disease. A small outbreak of MRSA was controlled by simple precautionary measures with no subsequent ongoing transmission of MRSA. PMID- 9874119 TI - A general practitioner STD training programme: meeting education and service provision needs. AB - This paper discusses the development, implementation and evaluation of a training programme in sexually transmitted diseases (STDs) for general practitioners (GPs) belonging to the Divisions of General Practice in the Sydney metropolitan area. The aims of the project were to involve GPs in the management of STDs, sexual health issues and the prevention of human immunodeficiency virus (HIV) infection and to ensure a consistent and high quality of care in the management of STDs and HIV. The strategy used was a two-stage programme consisting of a series of 3 educational modules (total 15 h) and an intensive clinical training programme for selected GPs for 12 times 4 h clinical sessions. Needs assessment of the participating GPs was conducted to ensure that the content of the theory modules was appropriate. Evaluation of the modules was conducted to examine both perceived quality of the actual presentations and changes in knowledge or skills as a result of participation in the project. PMID- 9874120 TI - HIV infection among patients with sexually transmitted diseases in rural South Africa. AB - A cross-sectional study of 360 patients presenting with sexually transmitted diseases (STDs) to a primary care clinic in KwaZulu/Natal, South Africa was done. Prevalence of HIV infection was 42.5%. HIV-infected patients were of similar age to uninfected patients (mean age 25.1 vs 26.1 years), but were less likely to be married (9.2% vs 18.8%; P=0.02). HIV prevalence was highest among young women (47.9% among women aged 15-34 years compared with 33.1% among men of the same age; P=0.03). History of a previous STD in the preceding 3 months was high (40.1% in HIV-infected patients). Similar proportions of the HIV-infected and HIV uninfected had sought care for the previous illness at private practitioners (16.9%), and primary care clinics (50.0%), and traditional healers (14.6%) or had treated themselves (18.5%). Patients with an STD are at very high risk of HIV infection in this setting. Repeat STDs are frequent and opportunities exist to improve treatment seeking behaviour, to reduce the risk of recurrent STDs, and hence to reduce the incidence of HIV infection. PMID- 9874121 TI - Changing patterns of HIV transmission and better targeting for intervention strategies. AB - We considered the HIV population of our area, comparing demographic characteristics between 2 consecutive 6-year periods to assess the current patterns of HIV transmission. All HIV-positive patients referred to our hospital from January 1985 to December 1996 were included in the study and were classified into 2 periods: A (January 1985 to December 1990) and B (anuary 1991 to December 1996). The variables analysed were: sex, age at first visit, HIV risk category. A total of 4284 HIV subjects were observed, 2306 in period A vs 1978 in period B (P=ns). Males were 76.3% vs 75.2% (P=ns). Mean age for males was 27.4 vs 32.4 years (P < 0.001) and for females 25.4 vs 30.1 years (P < 0.001). Intravenous drug users (IVDUs) were 88.4% vs 65.4% (P < 0.001), 'heterosexuals' 14.3% vs 24.8% (P < 0.001), 'men who have sex with men' 2.4% vs 4.8% (P < 0.001). Mean age by the main risk groups was: IVDUs 25.9 vs 29.7 years (P < 0.001); heterosexuals 30.4 vs 36 years (P=0.007); 'men who have sex with men' 35 vs 35 years. In conclusion, our study confirms the emerging role of heterosexuals in the current HIV epidemic. People older than teenagers seem to have misperceived their own risk of HIV infection, given the increase in the mean age occurred in the most recent years. This trend suggests the need for prevention strategies focusing more on heterosexual transmission and older people. PMID- 9874122 TI - Perceptions and patterns of reproductive tract infections in a young rural population in North-West Namibia. AB - Syndromic management of sexually transmitted diseases (STDs) is strongly promoted in Namibia. However, no data are available on rates of asymptomatic reproductive tract infections (RTIs) which would remain undetected and thus untreated in the general population. This study was aimed at assessing the prevalence of RTIs in a young rural population and to gain an insight into their health-care seeking behaviour for RTIs. A total of 97 patients (44 male, 53 female) <30 years of age attending the outpatient clinic of a rural hospital in Northern Namibia for problems unrelated to RTIs were interviewed and examined. Specimens were taken for investigation of RTIs according to a non-invasive diagnostic schedule. Laboratory examinations revealed 58 infections of the reproductive tract in 42 of the 97 patients (43%). In addition, HIV infection was detected in 13 cases (13%). RTI rates were highest in female teenagers (68%), but no infections were detected in their male counterparts. Of those 42 patients with a laboratory-diagnosed infection, 7 had both symptoms and signs (symptomatic), 16 had neither symptoms nor signs (asymptomatic), and 19 had signs on examination but no symptoms (inapparent). Awareness and perception of symptoms and signs of RTIs were low in the studied population. Provision of adequate treatment for RTIs alone is unlikely to have a major impact on the overall burden of disease since the largest burden of disease was found among those patients (predominantly females) with inapparent infections. Health education on RTIs including STDs and the symptoms they cause together with the importance of early attendance at health facilities and improved access to treatment need to be addressed urgently. PMID- 9874123 TI - Activity of thalidomide in AIDS-related Kaposi's sarcoma and correlation with HHV8 titre. AB - A phase II study of thalidomide was conducted to evaluate its efficacy and toxicity in the treatment of cutaneous AIDS-related Kaposi's sarcoma (AIDS-KS). To evaluate whether clinical response is correlated with titre of human herpesvirus 8 (HHV8) DNA in peripheral blood, levels were determined by serial end-point dilution at enrolment and 4-6 weeks later. Seventeen male HIV seropositive patients with histopathologically diagnosed KS were treated with thalidomide 100mg orally once nightly for 8 weeks. Response evaluation was performed using AIDS Clinical Trials Group (ACTG) criteria and analysis was by intention to treat. Six of 17 patients achieved a partial response (35%: 95% confidence interval 10-61%). Eight patients withdrew (6 owing to toxicity, one to early progression and one to non-compliance). HHV8 DNA load decreased by at least 3log10 to undetectable levels in 3 of the 5 virologically assessable partial responders. This preliminary study demonstrates that thalidomide has activity in the treatment of AIDS-KS and that clinical response is associated with a reduction of HHV8 DNA titre in peripheral blood. PMID- 9874124 TI - Risk profile of female sex workers who participate in a routine penicillin prophylaxis programme in Surabaya, Indonesia. AB - We conducted a sexually transmitted disease (STD) prevalence survey of 1867 female sex workers in Surabaya, Indonesia, some of whom reported participation in a routine penicillin prophylaxis programme. In Surabaya, 34% of female sex workers had received a prophylactic penicillin injection programme from the government within 28 days. Sex workers who had received routine prophylaxis injection were more likely to be less educated, to work in brothel complexes, and to have more customers per week than other sex workers. The prevalence rates of syphilis, gonorrhoea, chlamydia, and trichomoniasis were higher among sex workers who received the routine penicillin treatment than among those who had not received antibiotic treatment in the last 28 days. However, after adjustment for age, education, fee per sex act, number of customers, and condom use in the previous 7 days, only trichomoniasis was still significantly different (adjusted odds ratio of 3.2). High-risk women were more likely to participate in the routine penicillin prophylaxis programme. The lack of a demonstrable individual level protection from this prophylaxis treatment programme in this cross sectional study appears due to differential uptake of penicillin prophylaxis by women at higher presumptive risk for STD. Randomized clinical trials and mathematical modelling, together with observational data such as presented here, all can contribute to optimal understanding of a complex intervention like mass chemoprophylaxis for STD among female sex workers. PMID- 9874125 TI - Diagnostic accuracy of HIV-associated central nervous system toxoplasmosis. AB - Our objective was to examine the accuracy of diagnosis of HIV-associated central nervous system (CNS) toxoplasmosis. Individuals diagnosed with HIV-associated CNS toxoplasmosis and controls were ascertained from a population-based database. Diagnosis was confirmed by response to therapy or by histology. Symptoms, results of anti-Toxoplasma serology and use of Pneumocystis carinii pneumonia (PCP) prophylaxis were recorded. Central nervous system toxoplasmosis was confirmed in 54 (76%) of 75 patients. Reactive anti-Toxoplasma serology was associated with CNS toxoplasmosis (OR=20.4, 95% CI 3.1-175.8). Adjusting for CD4 and use of dapsone or aerosolized pentamidine, trimethoprim-sulphamethoxazole (TMP-SMX) for PCP prophylaxis was associated with lower likelihood of CNS toxoplasmosis (OR 0.3, 95% CI 0.1-0.7). Diagnosis of CNS toxoplasmosis is often incorrect. Another diagnosis is most likely in patients who are anti-Toxoplasma seronegative or who are receiving prophylactic TMP-SMX. PMID- 9874126 TI - Problems in physician's classification and reporting of congenital syphilis. AB - The diagnosis of congenital syphilis (CS) in newborns can only be made through a review of the mothers' testing and treatment history and through the infants' clinical and laboratory findings. We describe difficulties in the classification of CS by physicians and the health department during a recent syphilis epidemic. The records of infants identified as potential cases of CS by laboratory testing, discharge diagnosis, or health department records were reviewed by epidemiologists. The reasons for concordance and discordance in classification between the physician and the epidemiologist were determined. Congenital syphilis was identified in 126 infants. Seventeen cases were discordant and 12 cases concordant but the physician's classification was for incorrect reasons. Misclassification occurred because physicians lacked data known to the health department (n=7), health departments lacked data known to the physician (n=1), and physicians misinterpreted the case definition for CS (n=21). Suggestions for improving the diagnosis and reporting of CS are included. PMID- 9874127 TI - Human leukocyte derived interferon-alpha in a hydrophilic gel for the treatment of intravaginal warts in women: a placebo-controlled, double-blind study. AB - This placebo-controlled, double-blind study was aimed to evaluate the clinical efficacy, safety and tolerability of human leukocyte interferon-alpha (2 x 10(6) IU/g) incorporated in a hydrophilic gel (hydroxyethylcellulose, 1%) to cure intravaginal warts in women. Preselected, subjects (n=60) who ranged between 18 and 50 years of age (mean 23.7), harbouring 275 vaginal warts (mean 4.6) with clinical, histopathological and polymerase chain reaction (PCR) confirmed diagnosis of human papillomavirus (HPV) infections were randomly divided into 2 parallel groups. A precoded tube (45 g), active or placebo, with disposable applicators and instructions was given to each patient for one week's usage. Patients were demonstrated how to inject 4 g of the trial medication deep into the vagina 2 times daily for 5 consecutive days per week. During the 4-week treatment period, patients were examined on a weekly basis. Cure was defined as absence of clinical signs of infection, as well as PCR and Southern blot hybridization confirmed negative HPV DNA on molecular assay. By the cessation of the therapy 41.7% patients and 44.4% intravaginal warts were cured. Code disclosure revealed that interferon-alpha (2 x 10(6) IU/g) in gel had cured 73.3% patients, and 79.3% intravaginal warts, while placebo healed 10% patients and 8.1% lesions (active gel versus placebo; P<0.0001). Fifty-one patients (85%) complained of no drug-related adverse reactions. Nine patients (15%) mostly in the interferon-alpha gel experienced non-objective, mild headache, tenderness, with transient increase in their body temperature (>38 degrees C). In conclusion, the findings showed that along with non-objective mild side effects, human leukocyte interferon-alpha (2 x 10(6) IU/g) in a hydrophilic gel is significantly more effective than placebo to cure intravaginal warts in women. PMID- 9874128 TI - Highly active antiretroviral therapy (HAART) and prolonged survival of a patient with an HIV-related Burkitt lymphoma, despite an intracardiac relapse. PMID- 9874129 TI - The sonographic image of inflamed inguinal lymph nodes in primary syphilis. PMID- 9874130 TI - Chlamydia trachomatis as a probable cause of prostatitis. PMID- 9874131 TI - Website as a tool for patient education in sexually transmitted diseases. PMID- 9874132 TI - Provision of a dedicated 'men only clinic' (MOC) within a mixed sex GUM clinic increases uptake of sexual health services by men. PMID- 9874133 TI - Injector-inmates and anal sex with another man in prison. PMID- 9874134 TI - Use of the term 'Chlamydia'. PMID- 9874135 TI - A novel computerized system for analyzing motor and social behavior in groups of animals. AB - We present here the VMB Tracking System, a novel method for tracking locomotor activity, posture, thigmotactic scanning behavior and social interactions of up to eight animals at a time, at a high resolution (up to+/-0.1 mm). We used a commercially available computerized system that is considerably cheaper than other available methods. This system utilizes a basic personal computer controlling three transponders ('towers') fixed in space above the tested area, where animals as small as rats stroll freely in their normal habitat or in an experimental arena. Each tower emits infra-red (IR) pulses to a transponder ('button') adhered to a plastic mount glued to a shaved area of skin on the animal's back. When the button detects the IR pulses it responds with a button specific ultrasonic signal that is fed back to the towers. The 3D location of the buttons is calculated by triangulation. Movement parameters of each button, such as displacement trajectory, time, speed and acceleration, can be displayed on line and stored for off-line analysis. This system can be used to track animals in any lighting conditions, and to assess drug effects on the CNS, neuromuscular junction or muscle. As an example we demonstrate the ataxic effects of pentobarbital in rats. PMID- 9874136 TI - Precocious axons and improved survival of rat hippocampal neurons on lysine alanine polymer substrates. AB - We tested the hypothesis that other polymers of lysine would be better substrates for culture of CNS neurons than polylysine itself. In a serum-free medium optimized for survival of hippocampal neurons grown on substrates of poly-D lysine, 13% more neurons survived on substrates to which a sequential copolymer of lysine and alanine (LAS) was applied (P = 0.006). The effect was specific for the sequential polymer, in contrast to the random copolymer of lysine and alanine. This suggests that average cationic charge density is not as important as the spacing of these charges. More dramatically, immunostaining for the axon associated microtubule-associated protein, tau, indicated a 2-fold higher rate of fiber growth on LAS. The somatodendritic cytoskeletal component MAP2 also appeared to be increased in cells cultured on LAS. This suggests that cytoskeletal differentiation in general and axon formation in particular are stimulated by the LAS substrate. Scanning electron microscopy supported this conclusion. By circular dichroism, the conformation of LAS in phosphate-buffered saline appeared to be a random coil, indistinguishable from poly-D-lysine. These results indicate that LAS is a superior substrate to polylysine for growth of CNS neurons. LAS may be useful for regeneration of damaged circuits in the CNS as well as a substrate for connections to a neuroprosthesis. PMID- 9874137 TI - Drosophila nerve cord culture: a tool for studying neural development. AB - The culture of explanted neural tissues has been a useful tool for the study of cellular and molecular neurobiology in vertebrates. We have developed a technique for the culture of explanted ventral nerve cords from Drosophila embryos. We have examined the morphology and dynamic behaviour of the growth cones that extend from these nerve cords, and the effect of calcium deprivation on the bundling of axons that are regenerated from the explanted tissue. This technique offers a unique opportunity to combine in vitro techniques for neuronal cell culture with the powerful techniques of genetic analysis that are available with Drosophila. PMID- 9874138 TI - Micropipette system for sampling and injecting small volumes. AB - We have developed a micropipette system that allows sampling of preset volumes of liquids and injection of preset small amounts of liquids, using thermal expansion of an oil phase in a micropipette, achieved by increasing or decreasing the current applied to heat the shaft of the micropipette. The system has been tested to inject or collect volumes of about 20 nl, but smaller and larger volumes are possible. The amount injected or collected for a given temperature step of the micropipette is largely independent of the physical properties, e.g. of viscosity, into which the sample is injected or from which it is collected. The pipette tip is not measurably heated when heating the shaft for injection or collection, thus avoiding damage of the biological material into which the injection occurs or from which it is sampled. PMID- 9874139 TI - Flexible method to obtain high sensitivity, low-cost CCD cameras for video microscopy. AB - A simple method is described to extend image exposure times in video-rate CCD cameras and thereby, increase their sensitivity and reduce noise level of low light images. Most commercial video cameras lack the capability of extending image exposures since they operate regular television timing formats. The technique described here implements the control of the exposure times by selectively gating the image readout from the CCD sensor. This prevents the cyclic clearing of photo-charges occurring at regular video-rates, allowing image integration beyond the duration of single video field periods. Image readout is controlled by the duration of external gating pulses, giving the camera an efficient operational versatility under different light conditions. This technique is applicable to standard monochrome and color CCD cameras. The evaluations described here using this technique show that the light sensitivity of an standard video-rate CCD camera can be significantly improved, generating high quality images at low-light levels. These were comparable to those obtained with image intensifiers or intensified video cameras. Cameras are still compatible with regular video equipment, since this technique preserves the normal TV synchronization signals. Results in simulated and real experimental situations confirmed that this technique enables the use of affordable video-rate CCD cameras for a variety of fluorescence microscopy and optical recording applications. PMID- 9874140 TI - A simple, reproducible technique for establishing leptomeningeal tumors in nude rats. AB - The incidence of leptomeningeal carcinomatosis is on the rise and current treatment modalities have limited efficacy. The development of new treatment strategies has been hampered by the lack of an appropriate animal model that accurately parallels the clinical condition. We have developed a new surgical technique for the establishment of leptomeningeal tumors in rats. Our technique is simple, reproducible and associated with low morbidity and mortality. Tumor implantation resulted in a defined neurological endpoint and a reproducible time course of disease progression using both human medulloblastoma and breast carcinoma cell lines. This animal model provides an appropriate system for testing conventional and new biologic therapies in both early and late stages of leptomeningeal disease. PMID- 9874141 TI - Fluorescent tracers as potential candidates for double labeling of descending brain neurons in larval lamprey. AB - In larval lamprey, seven fluorescent tracers were tested as potential candidates for retrograde double labeling of descending brain neurons: Fluoro Gold (FG); fluorescein dextran amine (FDA); True Blue (TB); cascade blue dextran amine (CBDA); Fast Blue (FB); Texas red dextran amine (TRDA); and tetramethylrhodamine dextran amine (RDA). The first tracer (FG, TB, FB, or CBDA) was applied to the spinal cord at 40% body length (BL). In separate experiments, the second tracer (TRDA or RDA) was applied to the spinal cord at 20% BL. The tracer combination FG/TRDA was found to have the best optical properties for double labeling. However, application of FG to the spinal cord with the method used for the other tracers resulted in labeling of 'lateral cells' along the sides of the rhombencephalon that were presumed to be non-neuronal and that obscured some of the descending brain neurons. Control experiments suggested that FG was transported in the circulation to the brain, where the tracer was taken up by lateral cells. Therefore, a special technique was developed for applying FG to the spinal cord without allowing the tracer to enter the circulation. In larval lamprey, this important double-labeling technique that was developed for TRDA and FG is being used to examine axonal regeneration and projection patterns of descending brain neurons. PMID- 9874142 TI - Species-specific immunostaining of embryonic corneal nerves: techniques for inactivating endogenous peroxidases and demonstration of lateral diffusion of antibodies in the plane of the corneal stroma. AB - Species-specific and species-common monoclonal antibodies (MAbs) to nerve specific cell surface epitopes were used to compare pre-treatment techniques for nerve staining. Endogenous peroxidases were inactivated in four ways: (1) 0.3% hydrogen peroxide (H2O2); (2) 1% periodic acid (PA) (pH 1.85-1.95); (3) sodium meta-periodate (10-40 mM, pH 4.5); or (4) HCl (pH 1.80). Staining of chick and quail corneal nerves and dorsal root ganglia (DRG) nerves with the MAbs was species-specific. Staining of chick and quail corneal nerves was unaffected by pre-treatment with 0.3% H2O2, but was eliminated by pre-treatment with 1% PA. Chick and quail DRG nerve staining tolerated 0.3% H2O2, and at least one epitope also tolerated 1% PA. Corneal nerves of both chick and quail displayed concentration-dependent sensitivity to pre-treatment with sodium meta-periodate; DRG nerves were not sensitive to such pre-treatment. Corneal nerves tolerated pre treatment with HCI (pH 1.80), whereas DRG nerves did not. These findings indicate sensitivity of corneal nerve epitopes to oxidation, in contrast with sensitivity of DRG nerve epitopes to low pH. Results also indicate that tissue trimming regulated whole-mount staining of corneal nerves, suggesting that antibodies cannot diffuse across corneal basement membranes, even after detergent extraction. However, antibodies are able to diffuse laterally into the stroma from any cut edge. PMID- 9874144 TI - Rough annealing by two-step clustering, with application to neuronal signals. AB - To accomplish analyses on the properties of neuronal populations it is mandatory that each unit activity is identified within the overall noise background and the other unit signals merged in the same trace. The problem, addressed as a clustering one, is particularly difficult as no assumption can be made on the prior data distribution. We propose an algorithm that achieves this goal by a two phase agglomerative hierarchical clustering. First, an inflated estimation (overly) of the number of clusters is cast down and, by a maximum entropy principle (MEP) approach, is made to collapse towards an arrangement near natural ones. In the second step consecutive partitions are created by merging, two at time previously aggregated partitions, according to similarity criteria, in order to reveal a cluster solution. The procedure makes no assumptions about data distributions and guarantees high robustness with respect to noise. An application on real data out of multiple unit recordings from spinal cord neurons of mixed gas-anaesthetized rats is presented. PMID- 9874143 TI - Characterization of human joint proprioception by means of a threshold hunting paradigm. AB - A method has been developed to measure the proprioceptive performance of movement detection in human joints, with the threshold hunting paradigm commonly used in other sensory modalities. Methods applied in former studies regarding movement detection were very time consuming (hours), making them not applicable to patients. Using the threshold hunting paradigm, the subject changes the parameter value continuously around the threshold producing a threshold hunting curve, which itself might be valuable for investigation of movement detection performance. The method presented here, is very quick compared to other studies in this field, especially when velocity is hunted and the threshold values are very low. The threshold parameter value is stable since there is a control at any time over the subject's stimulus perception. Furthermore, a Monte Carlo simulation showed that hunting curves can clearly be distinguished from guessing. Also reliability is given. All these points allow to use the method to evaluate the proprioceptive performance of a single individual. Considering the results, the method described may be useful for basic and clinical investigations regarding the proprioceptive performance limits of movement detection aspects. PMID- 9874145 TI - A synthetic standard for competitive RT-PCR quantitation of 13 GABA receptor type A subunit mRNAs in rats and mice. AB - We describe a synthetic 769-bp DNA internal standard, GABARQuant 1, for measuring mRNAs of 13 GABA(A) receptor subunits by reverse transcriptase-polymerase chain reaction (RT-PCR). When it is transcribed into cRNA, added in known amounts to target mRNAs in extracts from rat or mouse tissue. competitively reverse transcribed into cDNA, and amplified by the polymerase chain reaction (PCR), the relative intensities of the amplified, stained target and standard DNA bands enable measurement of small amounts of mRNAs for GABA(A) receptor subunits alpha1 6, beta1-3, gamma1-3 and delta and the three cellular markers beta-actin, light neurofilament protein, and glutamine synthetase. For the subunits, most standard products (263-504 bp) differ in size from target products (398-564 bp) by 10-20%. Primer pairs span at least one intron, to prevent interference by genomic DNA, and at least one rat versus mouse restriction fragment length polymorphism (RFLP), to enable rat products to be distinguished from mouse products. PMID- 9874147 TI - Stereology of nerve cross sections. AB - Quantitative morphological data from nerve cross sections are of great value in experimental, developmental, and pathological studies. Efficient unbiased stereological techniques to estimate the total numbers and absolute size distributions of both myelinated and unmyelinated axons from a small sample of axons are described and evaluated. Axon numbers are estimated with the fractionator technique. Axon areas are estimated with the 2D-nucleator and the point counting technique for myelinated and unmyelinated axons, respectively. The axon perimeter is estimated by use of a line grid, and the myelin sheath thickness by direct orthogonal measurements in uniform, random locations. The empirically evaluated variance of the number estimates of myelinated axons was approximately approximately 0.05 when approximately 200 axons were counted. The empirical variance of area estimates obtained with a four-way 2D-nucleator was approximately 0.10 and contributed less than 0.5% to the observed total variance of axon areas within an animal. A light microscope modified for stereology with computer assisted stereological test systems and a motorised stage facilitate the acquisition of data for the myelinated axons, which is of primary interest in most studies. The unmyelinated axons are evaluated at the more time consuming electron microscopical level. PMID- 9874146 TI - Pre- or postembedding immunocytochemistry: which to choose for the localization of glial fibrillary acidic protein (GFAP)? AB - Immunostaining for glial fibrillary acidic protein was performed in the hippocampus and cerebellum of adult rats in order to compare the distributions of immunolabelling after pre- and postembedding procedures. The reactions of protoplasmic astrocytes and pericapillary astrocyte processes were investigated at the electron microscopic level. After the pre-embedding reaction, visualized with 3,3'-diaminobenzidine tetrahydrochloride, a granular precipitate was observed to decorate the rough endoplasmic reticulum of the astrocyte cell bodies and a precipitate filled the cytoplasm in the astrocyte processes. In studies with the postembedding procedure, immunogold particles were observed to be attached exclusively to the intermediate filaments of the astrocytic cytoskeleton both in the cell body and in the processes. It is concluded that the diaminobenzidine precipitate diffuses in the cytosol, pre-embedding immunocytochemistry is therefore, suitable for the overall labelling of astrocytes, whereas the postembedding procedure reveals the exact intracytoplasmic localization of glial fibrillary acidic protein. This explains the similar pre-embedding immunostaining patterns of astrocytes with markedly different amounts of glial filaments (e.g. protoplasmic, fibrous and reactive) and stresses the importance of the use of the postembedding method when an exact intracellular localization is required. The results also suggest that, in spite of claims of soluble cytoplasmic pools of this protein, the glial filaments are the exclusive domains of immunoreactive glial fibrillary acidic protein. PMID- 9874148 TI - Evaluation of peripheral nerve regeneration by nerve growth factor locally administered with a novel system. AB - An experimental model is presented for the local administration of neurotrophic substances at the site of peripheral nerve lesion. The model consists of a subcutaneously implanted silicone reservoir and a connecting tube with its distal end facing the severed and repaired nerve. Wistar rats (n = 180) were divided into two groups: a control group (saline-treated) (n = 90) and an NGF-treated group (n = 90). After sciatic nerve axotomy, an epineural repair was performed. NGF or saline were injected daily into the subcutaneous reservoir for the first 4 weeks after axotomy and weekly single dose between the 8th and 12th weeks. Both groups were divided into three subgroups of 30 animals each. The animals were sacrificed at 4, 8 and 12 weeks. Myelinated and non-myelinated axonal and thickness of myelin sheaths were quantified at the tibialis branch 25 mm distal to the nerve repair site. Axonal counts showed statistically significant differences between the treated and control groups at 4, 8 and 12 weeks. Finally, at 4 weeks the myelinated axons in the NGF group had significantly thicker myelin sheaths than in the control group. In comparison with other models of administration of different neurotrophic agents, NGF delivered through this system demonstrates a significant capacity for improving nerve regeneration without the problems inherent in multiple anesthesia, device exchange, or short half-life of the NGF single-dose administration. PMID- 9874149 TI - A non-radioactive in situ hybridization method that does not require RNAse-free conditions. AB - This report describes a quick and versatile method to perform non-radioactive in situ hybridization in which none of the hybridization steps are performed under RNAse-free conditions. This study demonstrates that in situ hybridization can be performed without an RNAse-free environment provided that the concentration of RNAse introduced during the experiment does not reach 0.1 microg/ml, a concentration that is unlikely to be achieved through an accidental contamination. Moreover, evidence is provided that the only step sensitive to RNAse degradation is the pretreatment since degradation during the hybridization step can not occur due to a very efficient protective effect exerted by formamide. Finally, our data suggest that endogenous RNAse activity might be readily neutralized through paraformaldehyde fixation. A feature of this method is the strong fixation that ensures a perfect tissue preservation, even at level of the fine structure of the cell processes. The method allows a uniform tissue penetration by sodium periodate and sodium borohydride treatment and can be easily used in combination with diaminobenzidine immunohistochemistry for double labeling experiments. PMID- 9874150 TI - A new method for the rapid and long term growth of human neural precursor cells. AB - A reliable source of human neural tissue would be of immense practical value to both neuroscientists and clinical neural transplantation trials. In this study, human precursor cells were isolated from the developing human cortex and, in the presence of both epidermal and fibroblast growth factor-2, grew in culture as sphere shaped clusters. Using traditional passaging techniques and culture mediums the rate of growth was extremely slow, and only a 12-fold expansion in total cell number could be achieved. However, when intact spheres were sectioned into quarters, rather than mechanically dissociated, cell cell contacts were maintained and cellular trauma minimised which permitted the rapid and continual growth of each individual quarter. Using this method we have achieved a 1.5 million-fold increase in precursor cell number over a period of less than 200 days. Upon differentiation by exposure to a substrate, cells migrated out from the spheres and formed a monolayer of astrocytes and neurons. No oligodendrocytes were found to develop from these human neural precursor cells at late passages when whole spheres were differentiated. This simple and novel culture method allows the rapid expansion of large numbers of non-transformed human neural precursor cells which may be of use in drug discovery, ex vivo gene therapy and clinical neural transplantation. PMID- 9874151 TI - Use of an AC induction motor system for producing finger movements in human subjects. AB - This report describes the set-up and evaluation of a novel system for producing precise finger movements, for tests of movement perception. The specifications were to construct a system using commercially available components that were easy to use but which offered both flexibility and also high precision control. The system was constructed around an industrial AC induction motor with an optical encoder, controlled by an AC servo digital control module that could be programmed using a simple, high-level language. This set-up fulfilled the requirements regarding position and velocity control for a range of movements and also the facility for the subject to move the joint voluntarily while still attached to the motor. However a number of problems were encountered, the most serious being the level of vibration and the inability to vary the torque during movements. The vibration was reduced to the point where it did not affect the subject, by the introduction of mechanical dampening using an anti-vibration coupling and a pneumatic splint. The torque control could not be modified during rotation and so the system could only be operated using constant torque for any given movement. PMID- 9874152 TI - Quantitative mapping analyzer for determining the distribution of neurochemicals in the human brain. AB - We developed a human brain mapping analyzer to determine the quantitative distribution of specific molecules, such as neurotransmitters or neuromodulators, based on a fluorescence microphotometry system that we had previously developed. The immunohistochemical fluorescence emitted from each microarea of a brain slice is collected into a photomultiplier tube through the pinhole and objective lens of a microscope. The brain slice is moved in the x- or y-direction by a motorized scanning stage under the objective lens, and the fluorescence intensities are measured quantitatively. The scanning speed is approximately 100 microareas/s, the maximum stage motion is 150 x 150 mm, and an unlimited amount of data can be gathered continuously by transfer to external memory devices. In this paper, this analyzer is characterized in detail, and the methods used for the preparation and analysis of human brain slices are described. As an example, the cholinergic distribution in hemispheric coronal slices of the adult human brain is analyzed. Each slice, immunohistochemically stained for choline acetyltransferase, was divided into approximately 3 million microareas (one area is 50 microm in diameter), and the distribution of the cholinergic neurons is shown. PMID- 9874153 TI - Image manipulation and error estimation for MRI analysis. AB - Manipulation of MRI images prior to volumetric analysis is a common practice that may unwittingly lead to errors in measurement. In this study, we examine the effects of two types of image manipulation: changes in the total number of slices used to obtain volume estimates (slice sampling rate) and image rotation. A phantom containing two regularly-shaped and two irregularly-shaped regions of interest (ROIs) was scanned using an SPGR sequence and 1-mm slices. Changes in slice sampling rate produced marked effects on volume estimation of irregularly shaped ROls. Comparatively little error was associated with changes in slice sampling rates for regularly-shaped ROIs. In addition, there was an interaction between image rotation in non-orthogonal planes and slice sampling rate. The data suggests that the ability to detect anatomical effects may be influenced by an investigator's choices concerning the number of slices included in a region of interest and image rotation when estimating volumes. PMID- 9874154 TI - Optical monitoring of PO2 changes and simultaneous recording of bioelectric activity in human and animal brain slices. AB - For investigations of hypoxic effects in nervous tissue, brain slices are often used as a model system. This provides the advantage that parameters of the micromilieu, e.g. pH and temperature can easily be controlled and measurements of different data, e.g. bioelectric potentials, ion activities etc. can be performed. It is of special importance that the PO2 the slice preparation is exposed to is equally controlled under these conditions. Therefore, a PO2 monitoring system is needed which provides representative values for the tissue environment. This requirement is fulfilled by an optical PO2 sensing method based on phosphorescence quenching as a function of PO2. Here, the application of this method as adapted for use in in vitro models is described and compared to the polarographic oxygen-sensing method. Both the optical and polarographic methods are comparable regarding accuracy and response time of measurements. Furthermore, both the optical method and electrophysiological measurements can be combined. Lastly, under experimental conditions, neither the phosphorescent dye Palladium meso-tetra-4-carboxyphenyl-porphine nor the illumination necessary for excitation of the dye influence bioelectric activity of neuronal tissue in vitro. In conclusion, the optical PO2 sensing method presented here provides a tool for reliable and continuous monitoring of PO2 in the immediate environment of brain slice preparations. PMID- 9874155 TI - Extended local access fibers: adjustable treatment of deep sites in the brain. AB - We describe a minimally invasive method for administering diffusible substances to computer-targeted regions of the primate brain. Treatments enter the brain by diffusion or under pressure from the portion of a curvilinear implant that lies within the region of interest. During the implantation surgery, a guide-tube cannula is passed through a cranial burr-hole and concentric, telescoping needles are extended from the cannula in sequence to trace a pre-planned course through brain tissue. After the leading end of the longest needle emerges through another burr-hole, the surgeon fastens a hollow dialysis fiber to its tip and draws the fiber into the brain by retracting the telescoping device in an orderly sequence. The surgery affects brain tissue only along the course of the fiber, causing about the same acute damage as the stereotactic introduction of an ordinary straight needle. The 'extended local access fiber' is relatively soft, flexible, and biocompatible. It remains permanently in the brain, with its semipermeable portion lodged in the target region. Leading and trailing portions are accessible from outside the brain. Experiments indicate a broad range of possible trajectories and confirm that substances delivered by access fiber can have physiological effects even 10 weeks after implantation. PMID- 9874156 TI - Pharmacological characterisation of dopamine overflow in the striatum of the normal and MPTP-treated common marmoset, studied in vivo using fast cyclic voltammetry, nomifensine and sulpiride. AB - The in vivo measurement of electrically-evoked dopamine overflow was measured for the first time in the striatum of control and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-treated common marmosets using fast cyclic voltammetry at untreated carbon-fibre microelectrodes, (7 microm, o.d.). The identity of dopamine was confirmed using electrochemical, pharmacological and histological criteria and complied with rat data from earlier studies. Dopamine overflow depended on the intensity, number of pulses, and frequency of the applied stimuli. Maximum dopamine overflow occurred using 1.0-2.0 mA, 200 micros pulse width, 150-200 pulses at 80-120 Hz stimulation of the medial forebrain bundle. Evoked dopamine overflow in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated marmosets showed a similar electrochemical and pharmacological profile compared to healthy controls, albeit the concentration detected was significantly reduced. The catecholamine uptake blocker, nomifensine, significantly increased the dopamine signal in control marmosets. However, in contrast, nomifensine had no significant effect on evoked dopamine overflow in 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-treated marmosets. Apart from demonstrating that fast cyclic voltammetry with electrical stimulation can be reliably used to monitor dopamine overflow within the primate brain, our results describe for the first time the technical prerequisites for the fast cyclic voltammetric technique in the non human primate brain. PMID- 9874157 TI - An in vivo method for studying afferent fibre activity from cervical paravertebral tissue during vertebral motion in anaesthetised cats. AB - We describe a method for characterizing and studying afferent input to the central nervous system arising from deep axial structures of the neck during defined cervical vertebral movement. Multiple or single unit recordings of afferent activity arising from identified receptive fields in these tissues can now be studied in situ while simultaneously inducing 'natural' stimulation of mechanoreceptors during well defined movements of the intact vertebral column. When combined with existing strategies for extracellular and intracellular recordings of neurones, the methods described here will allow in vivo investigation of the central effects of functionally identified afferents innervating identified receptive fields located in deep paravertebral tissues during a variety of discrete movements of individual vertebra. This has particular importance in determining the relative role that afferents innervating specific axial tissues have on identified neurones in the central nervous system. It will allow determination of the 'bias' of input to projection cells, such as 'hyperconvergent' neurones, during natural movement. Furthermore, it will allow investigation of their role in the control of somatic and autonomic reflex behaviour. PMID- 9874158 TI - Differential influence of a selective melanocortin MC4 receptor antagonist (HS014) on melanocortin-induced behavioral effects in rats. AB - We injected i.c.v. the natural agonist alpha-MSH (melanocyte-stimulating hormone) and the first selective melanocortin MC4 receptor antagonist HS014 (cyclic [AcCys11, D-Nal14, Cys18, Asp-NH(2)22]-beta-MSH(11-22) in rats and scored a number of behavioral effects which have been related to the melanocortic peptides. The results showed that HS014 (5 microg/rat) completely blocked alpha MSH (3 and 5 microg/rat)-induced grooming, yawning and stretching. Penile erections induced by alpha-MSH were, however, only partially blocked by HS014. Injections of alpha-MSH decreased food intake in food-deprived rats, whereas HS014 increased food intake. When the peptides were given together, the food intake was similar to that of saline treated controls. Locomotion/exploration and resting were not influenced by either peptide. Our data show that exogenous beta MSH decreases food intake, and that an endogenous central melanocortinergic inhibitory tone on feeding prevails which can be blocked with HS014, leading to an increase in food intake. Our data also provide evidence that grooming, stretching and yawning in rats may be mediated by the melanocortin MC4 receptor, whereas penile erections might perhaps be mediated by some other melanocortin receptor. PMID- 9874159 TI - Place conditioning of mice with the NMDA receptor antagonists, eliprodil and dizocilpine. AB - Effects of noncompetitive and competitive NMDA receptor antagonists have been repeatedly characterized using place conditioning models. The present study aimed to characterize the effects in mice of another NMDA receptor antagonist acting at polyamine binding site, eliprodil. Five-day conditioning with eliprodil (1-30 mg/kg, i.p.) resulted in a dose-dependent avoidance of an eliprodil-paired compartment during post-conditioning tests. These effects were: (i) observed both with eliprodil and without drug, and (ii) less pronounced in individually housed mice subjected to repeated social defeats and mild footshocks prior to and during the conditioning period (compared to group-housed and individually housed nonstressed mice). In a parallel set of experiments, the effects of dizocilpine (MK-801; 0.03-0.3 mg/kg, i.p.) were evaluated using the same study design as for eliprodil. Conditioned place preference was established with the dizocilpine dose of 0.3 mg/kg and this effect was not affected by housing/stressing or drug exposures during the test. PMID- 9874160 TI - Behavioural topography in the striatum: differential effects of quinpirole and D amphetamine microinjections. AB - Behavioural evidence has accumulated that supports the hypothesis that specific territories of the striatum contribute differentially to the control of motor behaviours. The present experiments compare the behavioural effects of microinjections of amphetamine (20 microg/0.5 microl) with those elicited by the D2-class dopamine receptor agonist quinpirole (3 microg/0.5 microl) following direct microinjection into three anatomically distinct sectors of the striatum: the nucleus accumbens, the ventrolateral striatum and the anterodorsal striatum. Our findings demonstrate that site-specific behavioural responses are induced by microinjections of amphetamine, but not of quinpirole, into the striatum. Our results suggest that widespread areas of the striatum are implicated in the induction of a syndrome of sedation, yawning and motor inhibition, observed readily following microinjections of quinpirole into the striatum. This evidence supports both homogeneity and segregation of function in the striatum at the behavioural level. Further, the results suggest that the elicitation of site specific action sequences at the level of the striatum seems to require cooperative interactions between D1-class and D2-class dopamine receptors. PMID- 9874161 TI - BIBP 3226 inhibition of nicotinic receptor mediated chromaffin cell secretion. AB - (R)-N 2-(diphenacetyl)-N-[(4-hydroxyphenyl)methyl]-argininamide (BIBP 3226) is a selective neuropeptide Y Y1 receptor antagonist with structural similarity to the C-terminal tripeptide of neuropeptide Y. Based on this similarity we questioned whether BIBP 3226 could act as an agonist. Incubation of BIBP 3226 with bovine chromaffin cells in culture results in the inhibition of nicotinic receptor stimulated catecholamine secretion (IC50 = 2.4 microM). The effect of BIBP 3226 is independent of neuropeptide Y action since the presence of neuropeptide Y in the culture medium does not alter the effect of BIBP 3226. BIBP 3226 decreased the efficacy of the nicotinic receptor agonist, 1,1-dimethyl-4-phenylpiperizinium (DMPP), but did not change its potency suggesting non-competitive inhibition. BIBP 3226 has a similar effect on nicotinic receptor-stimulated 45Ca2+ influx. BIBP 3226 does not inhibit [3H]norepinephrine release induced by high K+ and its effect is not pertussis toxin-sensitive. We conclude that not only can BIBP 3226 act as a neuropeptide Y receptor antagonist in bovine chromaffin cells but also act as an agonist and inhibit catecholamine secretion. PMID- 9874162 TI - Clozapine-induced dopamine levels in the rat striatum and nucleus accumbens are not affected by muscarinic antagonism. AB - The effect of the muscarinic antagonist, scopolamine, was examined for a change in the increase in extracellular dopamine, dihydroxyphenyl acetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxyindolacetic acid (5-HIAA), induced by haloperidol or clozapine in the striatum and nucleus accumbens of anaesthetised and awake rats, monitored using in vivo cerebral microdialysis. Rats received scopolamine (1 mg kg(-1); s.c.) or vehicle followed by haloperidol (1 mg kg(-1); s.c.) or clozapine (20 mg kg(-1); s.c.). Dopamine, DOPAC, HVA and 5-HIAA overflow into striatal or accumbens perfusates was determined using high performance liquid chromatography with electrochemical detection (HPLC-ECD). Scopolamine failed to modify the clozapine- or haloperidol-induced efflux of dopamine or its metabolites in either the striatum or nucleus accumbens following systemic administration in anaesthetised or awake rats. Although pretreatment with scopolamine tended to produce a smaller increase in the clozapine-induced efflux of DOPAC in striatal perfusates than following clozapine treatment alone, this was not statistically significant. Furthermore, local infusion of scopolamine (100 microM) with clozapine (1 mM) via the microdialysis probe did not attenuate the elevated efflux of dopamine observed following clozapine alone, in either the striatum or nucleus accumbens, in anaesthetised rats. This treatment did prevent the clozapine-induced increase in DOPAC and HVA in the striatum but not the nucleus accumbens. Carbachol (50 microM) infused into the dorsolateral striatum or nucleus accumbens raised extracellular dopamine levels 200% and 150%, respectively above baseline. Our data suggest that the increased efflux of dopamine and its metabolites in the rat basal ganglia following clozapine administration is not significantly dependent upon the interaction of clozapine with muscarinic receptors. PMID- 9874163 TI - Neuroprotective effects depend on the model of focal ischemia following middle cerebral artery occlusion. AB - The purpose of the present study was to compare the characteristics of the photochemical-induced thrombotic occlusion model and the thermocoagulated occlusion model of the middle cerebral artery in rats. We evaluated the neuroprotective effects of a NMDA receptor antagonist, (+)-MK-801 (dizocilpine, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cycloheptan-5,10-imine), an alpha-amino 3-hydroxy-5-methylisoxazole-4-propionic acid receptor antagonist, YM90K (6-(1H imidazol-1-yl)-7-nitro-2,3(1H,4H)-quinoxalinedione monohydrochloride), a Ca2+ channel antagonist, S-312-d (S-(+)-methyl-4,7-dihydro-3-isobutyl-6-methyl-4-(3 nitrophenyl)-thieno[2 ,3-b]pyridine-5-carboxylate), the radical scavengers, MCI 186 (3-methyl-1-phenyl-2-pyrazolin-5-one) and EPC-K1 (L-ascorbic acid 2-[3,4 dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyl-tridecyl)-2H-1-be nzopyran-6yl hydrogen phosphate] potassium salt), and a calcineurin inhibitor, FK506 (tacrolimus, Prograf). Although all tested agents in the present study attenuated the brain damage in the photochemical-induced thrombotic occlusion model, the radical scavengers did not attenuate the brain damage in the thermocoagulated occlusion model. The time course of brain damage and brain edema formation in the two models was examined. The time course of brain damage was not different in the two models, but the time course of brain edema was quite different. Brain edema formation in the photochemical-induced thrombotic occlusion model was significantly greater (P < 0.01) than that in the thermocoagulated occlusion model at all time point studied until 24 h after occlusion of the middle cerebral artery. The present study suggests that the photochemical-induced thrombotic occlusion model has characteristics of both permanent ischemia and ischemia reperfusion. PMID- 9874164 TI - GABA(B) receptors as potential targets for drugs able to prevent excessive excitatory amino acid transmission in the spinal cord. AB - The effects of GABA(B) receptor activation on the Ca2+-dependent depolarization induced overflow of endogenous glutamic acid and gamma-aminobutyric acid (GABA) was studied in rat spinal cord nerve terminals exposed in superfusion to 15 mM KCl. The GABA(B) receptor agonist (-)-baclofen inhibited the K+-evoked overflow of glutamate (EC50=0.098 microM) but was almost inactive against that of GABA. The overflow of both transmitters could be quite similarly inhibited by two other GABA(B) receptor agonists, 3-APPA (3-aminopropylphosphonous acid; EC50=0.087 and 0.050 microM in the case of GABA and glutamate, respectively) and CGP 44532 (3 amino-2(S)-hydroxypropyl)methylphosphinic acid; EC50=0.81 and 0.50 microM). The GABA(B) receptor antagonist CGP 35348 [3-amino-propyl(diethoxymethyl)phosphinic acid] blocked the effect of 3-APPA (1 microM) at the autoreceptors (IC50 approximately = 1 microM), but not at the heteroreceptors. In contrast, the effects of 3-APPA at both autoreceptors and heteroreceptors could be similarly prevented by another GABA(B) receptor antagonist, CGP 52432 [3-[[(3,4 dichlorophenyl)methyl]amino]propyl](diethoxymethyl) phosphinic acid (IC50 approximately = 10 microM). The data suggest that, in the spinal cord, GABA(B) autoreceptors on GABA-releasing terminals differ pharmacologically from GABA(B) heteroreceptors on glutamatergic terminals. Selective GABA(B) receptor ligands may be helpful for conditions characterized by excessive glutamatergic transmission in the spinal cord. PMID- 9874165 TI - In vivo modulation of rat hypothalamic histamine release by the histamine H3 receptor ligands, immepip and clobenpropit. Effects of intrahypothalamic and peripheral application. AB - We investigated the effect of the new potent and selective histamine H3 receptor agonist, immepip, and the histamine H3 receptor antagonist, clobenpropit, on in vivo neuronal histamine release from the anterior hypothalamic area of urethane anesthetized rats, using microdialysis. Intrahypothalamic perfusion with immepip at concentrations of 1 and 10 nM reduced histamine release to 75% and 35% of its basal level, respectively. Peripheral injection of immepip (5 mg/kg) caused a sustained decrease in histamine release of 50%. Clobenpropit potently increased histamine release after intrahypothalamic perfusion. The maximal increase in histamine release was 2-fold, observed at a concentration of 10 nM clobenpropit. Peripheral injection of clobenpropit (5-15 mg/kg) increased histamine release to about 150% of the basal value. A more marked increase in histamine release was found after injection of the histamine H3 receptor antagonist, thioperamide (5 mg/kg). In conclusion, intrahypothalamic perfusion of the histamine H3 receptor agonist, immepip and the histamine H3 receptor antagonist, clobenpropit, potently and oppositely modulated in vivo histamine release from the anterior hypothalamic area. The decreased histamine release after peripheral injection of immepip indicates that this novel agonist readily crosses the blood-brain barrier, making it a potential candidate for in vivo histamine H3 receptor studies. The differential increase in histamine release after peripheral injection of clobenpropit and thioperamide is discussed. PMID- 9874166 TI - Endothelium-mediated negative dromotropic effects of intravascular acetylcholine. AB - Acetylcholine acting through specific muscarinic membrane receptors causes a negative dromotropic effect and, in blood vessels, causes a vasodilation which results from its action on the endothelial cells via release of nitric oxide (NO). We decided to study this effect in isolated Krebs-Henseleit retrogradely perfused guinea pig hearts. A pair of stimulating electrodes was placed in the right atrium and to record the auricular-ventricular interval (A-V delay) one recording electrode was placed on the left atrium and the other on the tip of the ventricle. Hearts were paced at a rate of 3.8+/-0.1 Hz and perfused at a coronary flow rate of 9+/-0.25 ml/min. To obtain dose-response curves, single doses (as boluses) of acetylcholine were infused and the maximal A-V delay induced by each dose was determined. Perfusion of agents that inhibit NO accumulation (L-Arginine methyl ester (L-NAME) (0.5 mM)) or oxyhemoglobin (6 microM) caused displacement of the acetylcholine dose-response curve downward and to the right. Perfusion of NO-sparing agents like superoxide dismutase and dithiothreitol caused an upward and leftward displacement. Infusion of NO solutions or a NO donor (diethylamine nitric oxide [DEA-NO]) caused a dose-dependent negative dromotropic effect. In contrast, inhibition of the prostaglandin metabolic pathway by Indomethacin (0.01 mM) caused potentiation of acetylcholine effects which were reversed when it was co-perfused with L-NAME. When endothelial intravascular muscarinic receptors were selectively blocked by perfusion of a non-permeable macromolecule: dextran ( > 2000 kDa) covalently complexed to the receptor blocker (3-(2'-aminobenzhydryloxy) tropane)), the negative dromotropic effect of intravascular acetylcholine was diminished in a concentration-dependent manner up to complete blockade. Our data indicate that the dromotropic effect caused by intracoronary administration of acetylcholine is the result solely of activation of intravascular endothelial muscarinic receptors, that nitric oxide and prostaglandins are non-synergistic endothelial mediators of this effect and that there may be an interaction between NO and prostaglandin metabolic pathways. PMID- 9874167 TI - Atypical receptors mediate the response to endothelin-1 and sarafotoxin S6b in the human umbilical artery. AB - The receptors mediating smooth muscle response to endothelin-1 and sarafotoxin S6b in the human umbilical artery were investigated in vitro. Both agonists induced contractions that were unaffected by the endothelin ET(B) receptor antagonist BQ 788 (10(-9), 10(-8), 10(-7) M). The non-selective endothelin ET(A/B) receptor antagonist PD 142893 (10(-7) M) decreased the contraction induced by endothelin-1. PD 142893 (10(-9) M) enhanced the contraction induced by sarafotoxin S6b whereas higher concentrations had no effect. Removing the endothelium did not affect the antagonising action of PD 142893 on endothelin-1 induced contractions while the enhancement of the sarafotoxin S6b-induced contraction was abolished. Sarafotoxin S6b induced relaxation in segments precontracted by 5-hydroxytryptamine and exposed to the endothelin ET(A) receptor antagonist BQ 123 (10(-7) M) and PD 142893 (10(-9) M) abolished this relaxation. These endothelial receptors seem neither to be classical endothelin ET(A) nor endothelin ET(B) receptors and they are not activated by endothelin-1. PMID- 9874168 TI - Effect of muscimol on cholinomimetic-induced cardiovascular responses in rats. AB - Brain acetylcholine and gamma-aminobutyric acid (GABA) are both involved in the regulation of central cardiovascular control. Despite data from anatomical and electrophysiological experiments characterizing the interaction between central GABAergic and cholinergic neurotransmission, the potential significance of this interaction in central cardiovascular regulation remains unknown. The purpose of this study was to determine whether activation of GABA(A) receptors by intracerebroventricular or intrahypothalamic administration of muscimol affects the cholinergic agonist-induced cardiovascular responses. All experiments were performed in conscious, Sprague-Dawley rats instrumented with a guide cannula for drug injection and iliac arterial catheters for direct measurement of mean arterial pressure and heart rate. Administration of a cholinergic agonist, carbachol, either intracerebroventricularly or into the dorsomedial hypothalamic nucleus, produced a significant increase in mean arterial pressure, whereas injection of carbachol into the posterior hypothalamic nucleus caused a slight elevation in blood pressure. Pretreatment with muscimol 10 min before administration of carbachol prevented the carbachol-evoked blood pressure changes. On the other hand, carbachol produced variable changes in heart rate, depending on the site of injection. In [3H]quinuclydinyl benzilate binding experiments, muscimol did not displace the muscarinic radioligand from its binding sites, suggesting that it does not exert any direct antagonistic activity at muscarinic receptors. These results suggest that the dorsomedial hypothalamic nucleus is a potential site of action for microinjected carbachol and that the GABAergic system has an inhibitory influence on cholinergic neurons involved in blood pressure regulation. PMID- 9874170 TI - Capsaicin-induced desensitization is prevented by capsazepine but not by ruthenium red in guinea pig bronchi. AB - In isolated guinea pig bronchi, the influence of ruthenium red, capsazepine and extracellular Ca2+ on capsaicin-induced desensitization was examined to investigate whether this desensitization was mediated via a specific receptor coupled with an ion channel. Pre-exposure of tissues to capsaicin (1, 3 or 10 microM) caused a dose-dependent desensitization to the second application of capsaicin. However, the contractile responses to exogenous tachykinins were not changed after pre-exposure of tissues to capsaicin. This capsaicin-induced desensitization was prevented by capsazepine (30 microM), but not by ruthenium red added to tissues 20 min before pretreatment with capsaicin (3 microM). While the excitatory contractile response to capsaicin was markedly reduced in the absence of extracellular Ca2+, the desensitization induced by capsaicin was not changed by the removal of extracellular Ca2+. In summary, the results from the present study suggest that in vitro functional desensitization induced by capsaicin in guinea pig bronchi may involve changes in the vanilloid receptor and occur through a ruthenium red-insensitive pathway. PMID- 9874169 TI - Force, membrane potential and cytoplasmic Ca2+ responses to cyclic nucleotides in rat anococcygeus muscle. AB - Simultaneous recordings of membrane potential and force, and cytoplasmic calcium ([Ca2+]i) and force were made in rat anococcygeus to determine whether membrane hyperpolarisation plays a role in cyclic nucleotide-induced relaxation. In the presence of phenylephrine (0.2 microM), which evoked sustained contraction, an elevation in [Ca2+]i, and depolarisation, nitroprusside (5 microM) caused 96+/-3% relaxation, 77+/-3% decrease in suprabasal [Ca2+]i, and 16+/-2 mV hyperpolarisation. Forskolin (1 microM) caused 98+/-1% relaxation, 92+/-2% decrease in suprabasal [Ca2+]i, and 18+/-1 mV hyperpolarisation. These responses persisted in the presence of a variety of K+ channel blockers or in ouabain. The decrease in [Ca2+]i preceded the commencement of relaxation whereas the onset of hyperpolarisation lagged behind. Thus, cyclic nucleotide-mediated relaxation in rat anococcygeus is not dependent on hyperpolarisation mediated by the opening of K+ channels. Rather, it is suggested that the decrease in [Ca2+]i gives rise to hyperpolarisation, which reflects a decline in the Ca2+ dependent conductance(s) activated by phenylephrine. PMID- 9874171 TI - Mechanisms of carbachol-induced alterations in K+ transport across the rat colon. AB - The effect of carbachol, an agonist of the Ca2+ pathway, on K+ transport in rat proximal and distal colon was studied by measuring unidirectional fluxes, uptake, and efflux of Rb+, a marker for K+, in mucosa-submucosa preparations. Unidirectional ion flux measurements revealed that carbachol stimulated K+ secretion in the proximal colon by a marked increase in the serosa-to-mucosa flux (J(Rb)sm) and a more moderate rise in the mucosa-to-serosa flux (J(Rb)ms). In the distal colon carbachol had no effect on J(Rb)ms but J(Rb)sm was reduced after a transient increase finally resulting in an inhibition of K+ secretion. Carbachol caused a stimulation of mucosal Rb+ uptake in the distal colon, which was diminished in the presence of inhibitors of the apical H+-K+-ATPase, vanadate and ouabain. In contrast, in the proximal colon the serosal Rb+ uptake was enhanced by carbachol, an effect, which could be prevented by bumetanide, an inhibitor of the basolateral Na+-K+-2Cl(-)-cotransporter. Efflux experiments revealed that carbachol caused a transient increase of apical and basolateral Rb+ permeability in both colonic segments. In the distal colon, stimulated K+ efflux to the serosal side was reduced by quinine, efflux to the mucosal side was blocked by tetraethylammonium. In the proximal colon, carbachol-activated apical and basolateral K+ efflux were inhibited by Ba2+. In conclusion, these data suggest that in the distal colon carbachol stimulates the H+-K+-ATPase and the basolateral K+ efflux through quinine-sensitive K+ channels, whereas in the proximal colon carbachol induces K+ secretion due to a stimulation of the basolateral Na+-K+-2Cl(-)-cotransporter and an increased efflux to the luminal side via Ba2+-sensitive apical K+ channels. PMID- 9874172 TI - Effects of moguisteine on the cough reflex induced by afferent electrical stimulation of the superior laryngeal nerve in guinea pigs. AB - The study aimed to further demonstrate the peripheral antitussive properties of moguisteine. Firstly, the antitussive effect of moguisteine on the cough reflex induced by inhalation of citric acid aerosol was evaluated in conscious guinea pigs. Secondly, the effects of both moguisteine and codeine on the centrally mediated cough reflex induced by afferent electrical stimulation of the superior laryngeal nerve were investigated in anesthetized guinea pigs. Moguisteine (2.5 10 mg/kg, intravenously, i.v.) reduced the cough reflex induced by 7.5% citric acid aerosol in a dose-dependent manner, with an ED50 value of 0.55 mg/kg. Both i.v. (0.5-4 mg/kg) and intracerebroventricular (i.c.v., 5-20 microg) injection of codeine dose dependently inhibited the cough reflex induced by afferent electrical stimulation of the superior laryngeal nerve; the ED50 values were 0.91 mg/kg and 7.90 microg, respectively. The inhibitory effect of codeine (4 mg/kg i.v.) was abolished by pretreatment with naloxone (2 mg/kg intraperitoneally). In contrast to codeine, neither i.v. (4 and 20 mg/kg) nor i.c.v. (20 microg) injection of moguisteine affected the cough reflex. These results suggest that the antitussive effect of codeine is mediated by central opioid mechanisms, whereas the antitussive effect of moguisteine is mediated by peripheral mechanisms. PMID- 9874173 TI - Involvement of angiotensin II in development of spontaneous nephrosis in Dahl salt-sensitive rats. AB - We investigated the effect of angiotensin-converting enzyme inhibition on spontaneous nephrosis in Dahl salt-sensitive (Dahl/S) rats. Dahl/S rats fed on a normal sodium diet spontaneously developed nephrosis and mild hypertension from a young age. In young Dahl/S rats, an angiotensin-converting enzyme inhibitor, imidapril, attenuated the development of proteinuria accompanied by a decrease in blood pressure. Methylprednisolone, a potent therapeutic agent for proteinuria, did not affect the development of nephrosis. An angiotensin AT1 receptor antagonist, losartan, but not a Ca2+ channel blocker, verapamil, inhibited the development of nephrosis while both agents decreased blood pressure to a similar extent as imidapril. In mature Dahl/S rats, imidapril suppressed not only the development of proteinuria but also the glomerular lesions. It is concluded that the development of spontaneous nephrosis in Dahl/S rats is mediated by angiotensin II. PMID- 9874174 TI - Uridine triphosphate-sensitive pathway of Ca2+ release from the sarcoplasmic reticulum of rat skeletal muscle fibers. AB - The pyrimidine nucleotide, uridine triphosphate (UTP), was tested with skinned skeletal muscle fibers in order to investigate the UTP-sensitive pathway of Ca2+ release from the sarcoplasmic reticulum. The presence of ryanodine (200 microM), ruthenium red (10 microM) or heparin (2.5 mg/ml) did not affect the tension elicited in the presence of UTP, demonstrating that the UTP-induced Ca2+ release involved neither ryanodine nor inositol triphosphate-sensitive channels. Drugs such as compound 48/80 or cyclopiazonic acid used to inhibit Ca2+-ATPase in its reverse function appeared to be, respectively, non-specific or without any inhibitory effect on the tension induced by UTP. Finally, the UTP-induced tension as well as the trifluoperazine-induced tension were abolished in the presence of spermidine (50 mM), supporting the hypothesis that the UTP-sensitive pathway of the SR Ca2+ release might occur through the uncoupled calcium ATPase. PMID- 9874175 TI - Binding of YM158, a new dual antagonist for leukotriene D4 and thromboxane A2 receptors, to guinea pig lung membranes. AB - Arachidonic acid metabolites mediate inflammatory responses at a cellular level. The affinity of the newly synthesized compound YM158, 3-[(4-tert-butylthiazol-2 yl)methoxy]-5'-[3-(4-chlorobenzenesuf onyl)propyl-2'-(1H-tetrazol-5 ylmethoxy)benzanilide monosodium salt monohydrate, for leukotriene D4 and thromboxane A2 receptors was examined in radioligand binding assays. YM158 inhibited [3H]leukotriene D4 and [3H]U46619 (9,11-dideoxy-11alpha,9alpha epoxymethanoprostaglandin F2alpha) binding to guinea pig lung membrane preparations, with Ki values of 0.64+/-0.06 nM for leukotriene D4 and 5.0+/-0.88 nM for thromboxane A2 receptors. The Hill coefficients (nH) did not significantly differ from unity, indicating that this antagonism is competitive. In contrast, YM158 showed no affinity for several other receptors, including neurotransmitter related (alpha1-, alpha2-, beta-adrenoceptors, histamine, 5-HT, muscarinic, sigma), C5a, opioid, Ca2+ channel, K+ channel, protein kinase C, bradykinin, endothelin, neurokinin and platelet activating factor receptors. These studies indicate that YM158 is a highly selective dual antagonist for leukotriene D4 and thromboxane A2 receptors, and this has potential clinical and research applications. PMID- 9874176 TI - Angiotensin AT1 receptors in Clone 9 rat liver cells: Ca2+ signaling and c-fos expression. AB - In C9 (Clone 9) liver cells, angiotensin 11 increased the intracellular Ca2+ content, inositol phosphate production and c-fos mRNA expression. Other angiotensins were also active with the order of potency being angiotensin II = angiotensin III >> angiotensin I > angiotensin IV. Losartan, but not PD 123177 (1 (4-amino-3-methyl)-5-diphenylacetyl-4,5,6,7-tetrahydro-1H-imida zo [4,5c]pyridine 6-carboxylic acid), blocked the effects of angiotensin II. Pertussis toxin did not alter these actions of angiotensin II. These data indicate that the effects were mediated through angiotensin AT1 receptors involving pertussis toxin insensitive G-proteins. Phorbol myristate acetate was also able to increase c-fos mRNA expression. The action of angiotensin II was consistently greater than that of the active phorbol ester. Staurosporine but not genistein inhibited this effect of angiotensin II. Angiotensin II- and phorbol myristate acetate-induced proto-oncogene mRNA expression was attenuated in cells incubated overnight with the active phorbol ester, which suggests a major role of protein kinase C. PMID- 9874177 TI - Pharmacological profile of the rat intestinal crypt peptide YY receptor vs. the recombinant rat Y5 receptor. AB - Peptide YY and neuropeptide Y have potent antisecretory effects in rat small intestine. Scatchard analysis of [125I]peptide YY binding revealed a 10-fold higher concentration of receptors in rat jejunal crypt cells than in villus cells and no detectable receptors in colonic epithelium. Reverse transcription polymerase chain reaction analysis of neuropeptide Y Y5 receptor mRNA indicated that they are mainly expressed in rat jejunal crypts with very few or no expression in villus cells and colon epithelium, respectively. In order to determine whether neuropeptide Y Y5 receptors could represent the intestinal crypt receptor for peptide YY and neuropeptide Y, the ability of peptide YY, neuropeptide Y, pancreatic polypeptide and analogues to inhibit [125I]peptide YY binding to membrane prepared from rat crypt cells and COS-7 cells (African green monkey kidney cells) transfected with the rat neuropeptide Y Y5 receptor cDNA was tested. It appeared that several analogues displayed different inhibition constants (Ki) in the two binding assays, more especially N-alpha-acetyl-peptide YY-(22-36) which was 1200 x more potent in the crypt cell binding assay than in the recombinant neuropeptide Y Y5 receptor binding assay. These data support that the intestinal crypt peptide YY receptor is not a Y5 receptor. reserved. PMID- 9874178 TI - Ethanol inhibits mitogen activated protein kinase activity and growth of vascular smooth muscle cells in vitro. AB - The aim of this study was to determine the effect of ethanol on vascular smooth muscle cell proliferation and mitogen activated protein kinase (MAPK) signaling. Rat aortic smooth muscle cell growth in vitro was determined by measuring cell counts and [3H]thymidine incorporation. MAPK signaling was determined by assessing MEK (also referred to as MAPK kinase) activity by measuring phosphorylated extracellular signal-regulated kinase (pp44ERK - 1 and pp42ERK - 2) expression, and ERK activity by measuring ERK-2-dependent phosphorylation of myelin basic protein (MBP). In quiesced smooth muscle cells, ethanol treatment (24 h) inhibited serum-stimulated mitogenesis in a dose-dependent manner, (IC50 = 60 mM), in the absence of any effect on smooth muscle cell viability. In addition, ethanol treatment caused a significant shift to the right in the smooth muscle cell growth curve, extending the population doubling time from approximately 48 h (control) to approximately 70 h (ethanol). Acute (15 min) ethanol treatment reduced serum-stimulated pp44ERK - 1 and pp42ERK - 2 expression in a dose dependent fashion; 24.5+/-1.5% and 77.6+/-3.2% inhibition for 20 mM and 160 mM ethanol, respectively. Furthermore, there was a significant dose-dependent decrease in ERK2 activity in ethanol treated smooth muscle cells as compared to control smooth muscle cells. These data demonstrate an inhibitory effect of ethanol on smooth muscle cell proliferation and MAPK signalling in vitro. It is tempting to speculate that these actions of ethanol may contribute to its cardiovascular effects in vivo. PMID- 9874179 TI - Dietary beta-adrenoceptor agonists have a persistent effect on nitric oxide synthesis in rat cultured smooth muscle cells. AB - Several compounds including lipopolysaccharide and sympathomimetics stimulate the expression of the inducible nitric oxide synthase in vascular smooth muscle cells. We evaluated the effect of clenbuterol on nitric oxide (NO) production by vascular smooth muscle cells of the rat aorta in culture. Wistar rats were divided into three diet groups (control, clenbuterol and washout). Aortic vascular smooth muscle cells from rats from these 3 diet groups were cultured in the presence and absence of lipopolysaccharide and/or beta-adrenoceptor agonists. NO release was measured by Griess reagent. Clenbuterol or salbutamol added to cells from control rats potentiated lipopolysaccharide-induced NO release. Cells from rats fed on clenbuterol, in a medium without beta-adrenoceptor agonists, showed a similar potentiation, even after a 10-day washout period. The addition of beta-adrenoceptor agonists to the latter cells did not increase NO production. NG-Nitro-L-arginine decreased nitrite production in lipopolysaccharide-stimulated cells. Our results demonstrate that dietary clenbuterol has a persistent 'ex vivo' effect on lipopolysaccharide-induced NO production by cultured vascular smooth muscle cells. PMID- 9874180 TI - Intrathecal therapy of leptomeningeal CEM T-cell lymphoma in nude rats with anti CD7 ricin toxin A chain immunotoxin. AB - We have established a new xenogeneic animal model of leptomeningeal metastasis (LM) by intracisternal inoculation of human CEM T-cell lymphoma into nude rats, and used it to evaluate the anti-lymphoma efficacy of an anti-CD7 ricin A chain immunotoxin (DA7). In vitro incubation with 2 microg/ml DA7 for 72 h inhibited CEM cells by 90% in a trypan blue exclusion assay. To establish its anti-lymphoma activity, one and four days after cisternal inoculation of 10(6) CEM cells, eight animals each were treated cisternally with 10 microg DA7 in 50 microl PBS or sham treated with 50 microl PBS. Histopathologically, all eight sham-treated and five of eight DA7 treated animals showed typical features of LM with multilayers of tumor cells along the whole subarachnoid space and the ventricular walls, as well as subependymal and diffuse parenchymal tumor cell infiltration. Three DA7 treated animals were free of tumor. Two of these animals were asymptomatic long term survivors (> 90 days). The third tumor-free animal suddenly died on day 51. Histology revealed viral myocarditis. Median symptom-free survival was 51 days (range 29-90+ days) in DA7 treated and 34 days (range 29-87 days) in sham-treated animals (p = 0.12, log-rank test). Histologically, no signs of neurotoxicity or systemic toxicity was found. However, DA7 treated animals showed a tendency to a slower weight increase on days 6-28 after tumor cell inoculation. Our results indicate that this model is useful in studying leptomeningeal seeding and intracisternal treatment of lymphoma. The demonstrated anti-tumor effect of DA7 treatment deserves further evaluation especially regarding the application of DA7 in early stages of LM from T-cell lymphoma. PMID- 9874181 TI - Expression of multidrug resistance protein gene in patients with glioma after chemotherapy. AB - Two different ATP-binding membrane glycoproteins, the 170 kDa P-glycoprotein (P gp) and the 190 kDa multidrug resistance protein (MRP), are involved in the acquisition of multidrug resistance phenotypes in cancer cells. Overexpression of P-gp is often observed in various human tumors when treated with anticancer agents. In this study, we asked whether MRP was overexpressed in human gliomas after cancer chemotherapy. We investigated expression of MRP and P-gp before and after chemotherapy in tumor samples from patients with glioma. MRP expression was observed in 16 (70%) of 23 untreated patients, and the proportion of MRP-positive cells in the whole cell population ranged from 3 to 32% in the 16 MRP-positive patients. P-gp-positive tumors were observed in 4 (18%) of 23 patients, and the proportional rates of P-gp-positive cells in the whole cell population ranged from 4 to 23%. The proportional rate of MRP-positive or P-gp-positive glioma cells increased after chemotherapy when compared with that before chemotherapy in all patients examined. We could observe no statistically significant correlation between expression of MRP or P-gp and tumor grade. These results suggest that MRP as well as P-gp may be involved in acquired or intrinsic drug resistance in human glioma. PMID- 9874182 TI - Activation of the high affinity nerve growth factor receptor by two polyanionic chemotherapeutic agents: role in drug induced neurotoxicity. AB - Suramin is a polyanionic chemotherapeutic agent which causes severe peripheral neuropathy. The mechanisms of antineoplastic and neurotoxic activities are still poorly understood. Interference with growth factor receptor function has been suggested for suramin's chemotherapeutic function. Previous studies from our laboratory have demonstrated that suramin interfered with the function of nerve growth factor (NGF) and induced lysosomal storage defects within dorsal root ganglion neurons. Pentosan polysulfate (PPS) was used as another polyanionic agent, to compare these two cellular functions; NGF receptor interaction and disruption in glycolipid metabolism. Like suramin and NGF, PPS induced neurite outgrowth from the PC12 cell line which correlated with tyrosine phosphorylation of the high affinity NGF receptor (TrkA/gpl40) and ERK-1/MAP kinase. Ultrastructural studies of dorsal root ganglion exposed to PPS for various time periods were normal. This contrasted with suramin exposed cultures which consistently developed lamellar inclusion bodies (LIB) within 6 h. LIB formation with suramin treatment was associated with neuronal cell death, while PPS treatment did not cause any neurotoxic effects. These results indicated that PPS mimicked the effect of suramin on NGF receptors but did not cause similar accumulation of LIB. This suggested that the effect of polyanionic compounds on TrkA was not involved in LIB accumulation and subsequent induction of neurotoxicity. PMID- 9874183 TI - Effect of dietary vitamin A or N-acetylcysteine on ethylnitrosourea-induced rat gliomas. AB - It is our hypothesis that low grade gliomas are the glial counterparts of other precancerous lesions such as colon polyps and, therefore, suitable targets for chemoprevention. Steps in the molecular progression of gliomas have been described, indicating that an accumulation of abnormalities is required for progression to a high grade and interruption of this progression might be possible. An animal model of chemical glial carcinogenesis was used to test this hypothesis. Pregnant rats were injected intravenously with ENU (ethylnitrosourea) on the 18th day of gestation to induce gliomas in the offspring, which were randomized to receive control diet, diet supplemented with vitamin A palmitate, or diet supplemented with N-acetylcysteine. Animals exposed to ENU and receiving a control diet developed brain tumors and had a shortened life expectancy compared with rats unexposed to ENU. The animals treated with NAC showed no statistically significant delay in the time to tumor and no change in the histologic grade of the tumors when compared with animals receiving control diet, but the time to death from any cause of NAC treated animals differed significantly from untreated animals. Animals receiving high dose VA had statistically significantly prolonged time to tumor, survived significantly longer than untreated animals, but had no reduction in the total number of tumors or change in the histologic grade of their tumors. The theoretical basis of these results is likely due to the putative mechanism of action of these agents. These data indicate that glioma chemoprevention is possible and deserves further exploration. PMID- 9874184 TI - Acute leukemia following treatment of malignant glioma. AB - We report two patients with acute myeloid leukemia (AML) following therapy for malignant glioma; one was a young women treated heavily with alkylating agents for glioblastoma and the other a young man treated with high doses of procarbazine, lomustine, and vincristine (PCV) for anaplastic astrocytoma. We found 26 other examples of therapy related leukemia in adult and pediatric brain tumor patients. Including our two, there were 12 patients with malignant glioma; median interval from treatment to diagnosis of AML was 31 months. Nine adult malignant glioma patients all received nitrosoureas, some as the sole form of chemotherapy. No definite cases occurred after radiotherapy alone. Based upon analogy with other cancers, the cumulative dose of chemotherapy, especially alkylating agents, is the major risk factor for development of secondary AML. Agents implicated include carmustine (BCNU), lomustine (CCNU), and procarbazine. Conventional radiotherapy appears not to confer additional risk. Progressive macrocytosis, early dose reductions for thrombocytopenia, and refractory anemia may provide early diagnostic clues. Current glioma therapy is leukemogenic but the number of patients who survive the interval required to induce AML is small; nevertheless, the identification of chemosensitive types of glioma, and subgroups of patients who derive the most benefit from chemotherapy, may result in increasing numbers of patients at risk of long term complications. If regimens such as PCV continue to prove valuable in neurooncology the risk of leukemia will require integration into the clinical decision process. A search for more effective therapy with minimal mutagenicity remains critical. PMID- 9874185 TI - Cerebrospinal fluid ferritin in glioblastoma: evidence for tumor synthesis. AB - Ferritin concentrations in cerebrospinal fluid (CSF) of 16 patients with glioblastoma were found to be very high (mean, 103.0 ng/mL) relative to 29 patients with viral meningitis and 20 patients with headache (mean concentrations, 5.4 and 4.3 ng/mL respectively). Simultaneous measurement of ferritin in CSF and serum revealed CSF concentrations exceeding those in serum in 11 of 16 patients with glioblastoma (CSF vs. serum ratio, 132.6%), contrasting with very low ratios in patients with meningitis (7.9%) or headache (4.9%). Ferritin was detected immunocytochemically in tumor cell cytoplasm in a resected glioblastoma. Thus, ferritin in CSF appears to be produced by glioblastoma cells, with a biologic significance requiring further investigation. PMID- 9874186 TI - Ependymomas: MIB-1 proliferation index and survival. AB - The biologic behavior of ependymomas is highly variable, and its correlation with histologic features is at best imprecise. This retrospective study attempted to correlate the malignant histologic characteristics of ependymomas with MIB-1 proliferation index and survival. Biopsy and resection specimens taken from 34 patients who received treatment 1972 to 1996 were histologically examined. The patients' ages range was 1 to 59 years. The histologic specimens were assessed for anaplastic features (necrosis, mitosis, vascular proliferation, cellular pleomorphism, and overlapping of nuclei) and an MIB-1 (Ki-67 antigen) proliferation index was also determined. The overall median MIB-1 proliferation index was 7.8% (range 0.1 - 62.5%). An MIB-1 of 20% was significant for a decrease in survival (RR = 5.7) (p = 0.0013). The median MIB-1 for patients < 20 years old was 20.6% with range (0.1, 43%), while that for patients > 20 years was 5.1% (range 0.2, 9.4%) (KW p = 0.055). Three of 5 histological features evaluated were significantly associated with outcome: > 5 mitotic figures per high-power field, necrosis, and vascular proliferation, but not nuclear overlap or pleomorphism. All pathologic factors except pleomorphism were significantly related to the MIB-1 proliferation index. In brief, our data support the association of poor prognoses in ependymomas with young age, the presence of three to four anaplastic histologic features, and an MIB-1 proliferation index > 20%. PMID- 9874187 TI - Astroblastoma: does histology predict biologic behavior? AB - Astroblastoma is a rare and controversial tumor about which little is known. We have made the diagnosis in seven patients since 1990 at Memorial Sloan-Kettering Cancer Center. Four patients had astroblastomas with anaplastic features, whereas three patients had tumors which were well-differentiated. All three patients with low grade lesions are alive and recurrence free after surgery alone (mean follow up 29 months). All four patients with anaplastic astroblastoma were treated with surgery, radiotherapy and chemotherapy. One died of infection during induction chemotherapy. Two others died of tumor progression at 28 and 42 months. Radiographic response to chemotherapy was seen in one patient. The results of our series and other reports suggest that anaplastic histology is a prognostic factor in the setting of astroblastoma. More aggressive treatment is necessary for patients with anaplastic astroblastoma although the precise role of irradiation and chemotherapy cannot be defined at this time. PMID- 9874188 TI - Preoperative superselective embolization of skull-base meningiomas: indications and limitations. AB - We evaluated the clinical significance of preoperative superselective embolization for skull-base meningiomas. The subjects consisted of 20 patients with skull-base meningiomas, and were classified into a preoperative embolized group and a non-embolized group. The volume of blood transfused during the operation, the length of the operative procedure and the neurological outcome were compared between the two groups. The results showed that, in tumors smaller than 6 cm, the blood lost during the operation was significantly less in the embolized group. In tumors larger than 6 cm, there was not difference in blood lost, perhaps because larger meningiomas tend to have tiny blood vessels that are unsuitable for preoperative embolization. There was no difference in the length of the operation between the two groups. The embolized group tended to show a better clinical outcome than the non-embolized group. PMID- 9874189 TI - Patient selection criteria for the treatment of brain metastases with stereotactic radiosurgery. AB - In this study we evaluate prognostic factors that predict local-regional control and survival following stereotactic radiosurgery (SRS) in patients with brain metastasis and establish guidelines for patient selection. Our evaluation is based on 73 patients with brain metastasis treated with SRS at the University of Minnesota between March 1991 and November 1995. The ability of stereotactic radiosurgery to improve local control in patients with brain metastases is confirmed in our study in which only 6 of 62 patients failed locally after SRS, with an actuarial local progression-free survival of 80% at 2 years. Variables that predicted worse prognosis were larger tumor size (p = 0.05) for local progression-free survival and multiplicity of metastasis (p = 0.03) and infratentorial location of metastases (p = 0.006) for regional progression-free survival. Absence of extracranial disease, KPS > or = 70, and single intracranial metastasis were significant predictors of longer survival. Patients who fulfill all three criteria will survive longer after SRS (MS = 17.7 months) and will most likely benefit from the increase local control in the brain achieved by SRS. Survival in patients who do not meet any of these criteria is very poor (MS = 1.5 months), and these patients are less likely to benefit from this treatment. Careful selection of patients for SRS is warranted. PMID- 9874191 TI - High dose chemotherapy for childhood ependymona. PMID- 9874190 TI - Intracranial osteosarcomas: report of four cases and review of the literature. AB - Four cases of intracranial osteosarcoma are reported. In three cases the tumors were primary neoplasms arising from the sellar region, base of the occiput and the parieto-occipital area. One case represented osteosarcoma metastatic to the cerebellum, a site not previously described, from a primary tumor at the femur. In addition, the current literature on intracranial osteosarcoma is reviewed. PMID- 9874192 TI - Mechanisms of control of heart glycolysis. AB - This review focuses on the mechanisms of control of heart glycolysis under conditions of normal and reduced oxygen supply. The kinetic properties and the biochemical characteristics of control steps (glucose transporters, hexokinase, glycogen phosphorylase and phosphofructokinases) in the heart are reviewed in the light of recent findings and are considered together to explain the control of glycolysis by substrate supply and availability, energy demand, oxygen deprivation and hormones. The role of fructose 2,6-bisphosphate in the control of glycolysis is analysed in detail. This regulator participates in the stimulation of heart glycolysis in response to glucose, workload, insulin and adrenaline, and it decreases the glycolytic flux when alternative fuels are oxidized. Fructose 2,6-bisphosphate integrates information from various metabolic and signalling pathways and acts as a glycolytic signal. Moreover, a hierarchy in the control of glycolysis occurs and is evidenced in the presence of adrenaline or cyclic AMP, which relieve the inhibition of glycolysis by alternative fuels and stimulate fatty acid oxidation. Insulin and glucose also stimulate glycolysis, but inhibit fatty acid oxidation. The mechanisms of control underlying this fuel selection are discussed. Finally, the study of the metabolic adaptation of glucose metabolism to oxygen deprivation revealed the implication of nitric oxide and cyclic GMP in the control of heart glucose metabolism. PMID- 9874193 TI - The deletion of the C-terminal tail and addition of an endoplasmic reticulum targeting signal to Alzheimer's amyloid precursor protein change its localization, secretion, and intracellular proteolysis. AB - The metabolic pathway of Alzheimer's amyloid precursor protein (APP) involves restricted intracellular proteolysis by secretases, which leads to the secretion of the N-terminal soluble APP (sAPP) and the generation of a cell-associated C terminal fragment. The precise cellular sites at which these processes occur remain unknown. In this report, we describe the route of APP sorting and the processing site using novel systems with and without sorting signals on the APP molecule. One system involves the replacement of the C-terminal ten amino acids of APP with Adenoviral E19 protein containing an endoplasmic reticulum (ER) retrieval signal (APPE19); the other involves deleting the last ten amino acids corresponding to the replaced site (APPdeltaC10). APPE19 localized mainly within the cis/medial Golgi compartment and exclusively suppresses the secretion of APP. In contrast, deletion of the C-terminal tail promotes sAPP secretion by a constitutive secretion pathway. Metabolic labeling followed by immunoprecipitation with anti-APP antibody revealed that APPE19 is rapidly degraded within 30 min and that the subsequent intracellular turnover rate is decreased with 40% of the protein retained within the cells even after a chase period a 3 h. In contrast, APPdeltaC10 is rapidly eliminated from the intracellular compartments and secreted into the culture medium. The surface internalization and recycling processes of this protein are relatively impaired compared with wild-type APP. The ratios of the levels of production to secretion of sAPP alpha, the N-terminal, soluble APP fragment released by alpha-secretase, are proportional to the secretion efficiencies among APP species, suggesting the localization of alpha-secretase within a compartment late in the constitutive secretion pathway. These secretion mutants which utilize ER targeting signals are useful tools for analyzing the location of secretases and the intracellular degradation system within a constitutive secretion pathway such as ER quality control. PMID- 9874194 TI - The solution NMR structure of a blue-green algae hepatotoxin, microcystin-RR--a comparison with the structure of microcystin-LR. AB - The microcystin-RR structures are compared with the structures of microcystin-LR in solution as well as in the crystal structure of the complex with protein phosphatase. The gross structures of the two peptides are similar, but with a more accentuated and compact saddle structure for microcystin-RR. The structural differences affect the hydrogen-bond pattern in the peptides and the location of the side chain of N-methyldehydroalanine, both of which are important for the ability of the peptide to form a tight complex with protein phosphatase. These structural differences may contribute to the observed differences in toxicity of microcystin-RR and microcystin-LR. PMID- 9874195 TI - Inhibition of Streptomyces griseus aminopeptidase and effects of calcium ions on catalysis and binding--comparisons with the homologous enzyme Aeromonas proteolytica aminopeptidase. AB - Streptomyces griseus aminopeptidase is a zinc metalloenzyme containing 2 mol zinc/mol protein, similar to the homologous enzyme Aeromonas proteolytica aminopeptidase. In addition, a unique Ca2+-binding site has been identified in the Streptomyces enzyme, which is absent in the Aeromonas enzyme. Binding of Ca2+ enhances stability of the Streptomyces enzyme and modulates its activity and affinity towards substrates and inhibitors in a structure-dependent manner. Among the three hydrophobic 4-nitroanilides of alanine, valine and leucine, the latter displays the largest overall activation (increase in k(cat)/Km). Large enhancements in affinity (1/Ki) upon Ca2+ binding have been observed for inhibitors with flexible (leucine-like) residues at their N-termini and smaller enhancements for inhibitors with rigid (phenylalanine-like) residues. PMID- 9874196 TI - Site-directed mutagenesis of catalytic residues of influenza virus neuraminidase as an aid to drug design. AB - The neuraminidase of influenza virus is a surface glycoprotein that catalyzes the hydrolysis of glycosidic linkages between terminal sialic acids and adjacent sugar moieties. Neuraminidase function is critical for the spread of virus to new cells, and if the enzyme activity is inhibited, then virus infection is abrogated. The neuraminidase active site is conserved in all influenza type-A and type-B isolates, which makes it an excellent target for drug design. To determine the potential for resistance to develop against neuraminidase inhibitors, we have constructed mutations in seven of the conserved active-site residues of a type B (B/Lee/40) neuraminidase and analyzed the effect of the altered side chains on enzyme activity. There is a reduction in k(cat) in all our mutants. A transition state analogue inhibitor shows variation in Ki with the mutant neuraminidases, allowing interpretation of the effects of mutation in terms of transition-state binding and product release. The results show that Tyr409 is the most critical residue for enzyme activity, but that Asp149, Arg223, Glu275 and Arg374 also play important roles in enzyme catalysis. Based on the pH profile of neuraminidase activity of the D149E mutant protein, we conclude that Asp149 is not a proton donor, but is involved in stabilizing the transition state. If designed inhibitors are targeted to these residues where mutations are highly deleterious, particularly Tyr409, Glu275 and Asp149, the virus is unlikely to generate resistance to the drug. PMID- 9874197 TI - Identification and characterization of human PMP34, a protein closely related to the peroxisomal integral membrane protein PMP47 of Candida boidinii. AB - The cDNA of a human protein displaying 33% identity to the peroxisomal integral membrane protein PMP47 of Candida boidinii (CbPMP47) was cloned. It encodes a protein of 307 amino acids, with a calculated molecular mass of 34,566 Da. The protein, which we name HsPMP34, contains six membrane spans and belongs, like PMP47, to the family of mitochondrial solute carriers. Based on overlapping available expressed sequence tags (ESTs) combined with the sequencing of one EST, the amino acid sequence of the mouse counterpart (molecular mass of 34,422 Da) could also be deduced. By means of reverse-transcription PCR, the presence of PMP34 mRNA was revealed in liver of man and in liver and other tissues of mouse and rat. The human gene is localized to chromosome 22q13. Given the similarity of HsPMP34 not only to CbPMP47 but also to known or putative mitochondrial transporters, its subcellular localization was investigated. Fluorescence microscopy of HepG2 cells or mouse fibroblasts transfected with a plasmid encoding a HsPMP34/green fluorescent fusion protein revealed a punctate pattern, reminiscent of a peroxisomal distribution. By indirect immunofluorescence, the HsPMP34/green fluorescent fusion protein clearly colocalized with the dimeric peroxisomal thiolase. Upon transfection of mouse fibroblasts lacking functional peroxisomes due the knock-out of PEX5 [Baes, M., Gressens, P., Baumgart, E., Carmeliet, P., Casteels, M., Fransen, M., Evrard, P., Fahimi, D., Declercq, P. E., Collen, D., Van Veldhoven, P. P. & Mannaerts, G. P. (1997) Nat. Genet. 17, 49 53], fluorescence was associated with larger and less-abundant structures. Taken together, the data indicate that HsPMP34 is a peroxisomal membrane protein and is either the human counterpart of CbPMP47 or closely related to it. According to its structure, the protein is most probably involved in transport. PMID- 9874198 TI - Transducin-like Enhancer of split 2, a mammalian homologue of Drosophila Groucho, acts as a transcriptional repressor, interacts with Hairy/Enhancer of split proteins, and is expressed during neuronal development. AB - Groucho is a Drosophila transcriptional repressor involved in neurogenesis, segmentation, and sex determination together with basic helix-loop-helix proteins of the Hairy/Enhancer of split (HES) family. Several mammalian Groucho homologues, the Transducin-like Enhancer of split (TLE) 1 through 4 proteins, share similar properties with their Drosophila counterpart, suggesting that TLE proteins perform functions analogous to the roles of Groucho in Drosophila. The aim of this study was to examine this possibility by characterizing the properties of TLE2 and extending the analysis of TLE1. It is shown here that TLE2 and TLE1 are transcriptional repressors that contain two separate repression domains, located either within a Gln-rich amino terminal region or within an internal domain characterized by an abundance of Ser, Thr, and Pro residues. In addition, both TLE2 and TLE1 can homo- and heterodimerize through a short region that is part of their amino-terminal transcription repression domains. Finally, TLE2 interacts and is co-expressed with mammalian HES proteins in both neural and non-neural tissues. Taken together, these findings implicate TLE2 in transcriptional repression and define the structural elements that mediate transcriptional and protein-protein interaction functions of Groucho/TLE proteins. PMID- 9874199 TI - Degradation of the morpholino ring in the crystal structure of cyanomorpholinodoxorubicin complexed with d(CGATCG). AB - The cyanomorpholino analogue of antitumor anthracycline doxorubicin possesses an intense potency and differs from the parent compound in cross-resistance and other biological properties. The induction by cyanomorpholinodoxorubicin of both DNA cross-links and strand scission suggests an altered mode of action relative to doxorubicin with a different DNA-interacting capacity. We have co-crystallized 3'-(3-cyano-4-morpholinyl)-3'-desaminodoxorubicin (CMD) with the DNA hexamer d(CGATCG) and have determined the crystal structure at 0.16-nm resolution. The complex crystallizes in the space group P4(1)2(1)2 and is similar to the previously reported anthracycline/DNA structures, with the drug intercalated at the CpG step, forming hydrogen bonds with the guanine residue. The structure reveals that the morpholino moiety has undergone a major rearrangement with loss of the cyano group and opening of the morpholino ring. The compound actually bound to DNA in the complex resembles N-(2-hydroxyethyl)doxorubicin, which was previously identified as a hydrolysis product of CMD. No DNA alkylation has been observed. However, the structure shows that the active site of the morpholino ring, after dissociation of the cyano group, lies in the minor groove in proximity of the A/T base pair. This may indicate that a C/G base pair next to the intercalation site, with NH2 group in the minor groove, is required for DNA alkylation. PMID- 9874200 TI - Structural organization and chromosomal localization of the human hepatocyte growth factor activator gene--phylogenetic and functional relationship with blood coagulation factor XII, urokinase, and tissue-type plasminogen activator. AB - The organization and structure of the gene coding for hepatocyte growth factor activator (HGFA) have been determined by isolation of unique clones from a human genomic library. These clones were characterized by restriction mapping, Southern blotting and DNA sequencing. The complete sequence of the gene was determined and found to span about 7.5 kilobases of DNA and consist of 14 exons separated by 13 introns. The coding region of HGFA consists of multiple putative domains that are homologous to those observed in blood coagulation factor XII (FXII). These regions were found as separate exons in the gene, and the exon/intron arrangement was similar to that of FXII, suggesting that the genes for HGFA and FXII have arisen through gene duplication events from a common ancestral gene. The major transcription initiation site is located 75 bp upstream of the translational start codon. The gene was mapped to chromosome 4p16, using spot-blot hybridization on sorted chromosomes and fluorescence in situ hybridization on metaphase chromosome spreads. The phylogenetic and functional relationships between HGFA and FXII as well as urokinase and tissue-type plasminogen activator are discussed. PMID- 9874201 TI - X-ray crystal structure of the Desulfovibrio vulgaris (Hildenborough) apoflavodoxin-riboflavin complex. AB - The apoprotein of flavodoxin from Desulfovibrio vulgaris forms a complex with riboflavin. The ability to bind riboflavin distinguishes this flavodoxin from other short-chain flavodoxins which require the phosphate of FMN for flavin binding. The redox potential of the semiquinone/hydroquinone couple of the bound riboflavin is 180 mV less negative than the corresponding complex with FMN. To elucidate the binding of riboflavin, the complex has been crystallized and the crystal structure solved by molecular replacement using native flavodoxin as a search model to a resolution of 0.183 nm. Compared to the FMN complex, the hydrogen-bonding network at the isoalloxazine sub-site of the riboflavin complex is severely disrupted by movement of the loop residues Ser58-Ile64 (60-loop) which contact the isoalloxazine by over 0.35 nm, and by a small displacement of the isoalloxazine moiety. The 60-loop movement away from the flavin increases the solvent exposure of the flavin-binding site. The conformation of the site at which 5'-phosphate of FMN normally binds is similar in the two complexes, but in the riboflavin complex a sulphate or phosphate ion from the crystallization buffer occupies the space. This causes small structural perturbations in the phosphate-binding site. The flexibility of the 60-loop in D. vulgaris flavodoxin appears to be a contributing factor to the binding of riboflavin by the apoprotein, and a feature that distinguishes the protein from other 'short chain' flavodoxins. In the absence of the terminal phosphate group, free movement at the 5'-OH group of the ribityl chain can occur. Thus, the 5'-phosphate of FMN secures the cofactor at the binding site and positions it optimally. The structural changes which occur in the 60-loop in the riboflavin complex probably account for most of the positive shift that is observed in the midpoint potential of the semiquinone/hydroquinone couple of the riboflavin complex compared to that of the FMN complex. PMID- 9874202 TI - The high degree of internal flexibility observed for an oligomannose oligosaccharide does not alter the overall topology of the molecule. AB - The conformational properties of oligosaccharides are important in determining their biological properties, such as recognition by proteins. The structural and dynamic properties of many oligosaccharides are poorly understood both because of a lack of experimental data (usually obtained from solution NMR parameters) and because of gross approximations frequently invoked in theoretical models. To characterise the oligomannose oligosaccharide Man,GlcNAc2 we have acquired a more extensive NMR data set and performed the first unrestrained molecular dynamics (MD) simulation in water of this large oligosaccharide (employing the GLYCAM_93 parameter set with the AMBER force field). Good agreement is seen between the computed dynamics data and the results of both an isolated spin pair (ISPA) analysis of short mixing time NOE data and NOE build-up curves for mixing times from 100 to 2000 ms. The number of experimental conformational constraints obtained in this study are in principle sufficient to fully define a rigid structure. The fact that this could not be done indicates a high degree of internal flexibility and/or the presence of multiple conformations about the glycosidic linkages. Independently, the same conclusions are reached from an analysis of the MD results. In addition, the theoretical results allow the overall topology of the molecule and its intra-molecular and solvent-mediated hydrogen bonding pattern to be defined. Extensive re-organisation of solvent and inter-residue hydrogen bonds is shown to be required for significant conformational changes to occur, resulting in relatively long life-times for distinct glycosidic linkage conformations, despite the high local flexibility of the glycosidic linkages. This factor is also seen in the overall topology of the molecule, where the considerable internal flexibility is not translated into gross changes in structure. The control exerted by the solvent over both the flexibility and overall topology of an oligosaccharide has important implications for recognition processes and for the conformational properties of glycans attached to glycoproteins. PMID- 9874203 TI - Rejoining of DNA double-strand breaks in vitro by single-strand annealing. AB - Nonhomologous DNA end joining (NHEJ) is considered the major pathway of double strand break (DSB) repair in vertebrate cells. Various studies indicated the existence of at least two different NHEJ pathways; one that joins DNA ends accurately and depends on Ku, a protein heterodimer that binds to DNA ends, and one that generates deletions and is independent of Ku. While the former pathway has been characterised in some detail, only little is known about the latter error-prone. We have partially purified such an NHEJ activity from extracts of Xenopus laevis eggs. End-joined junctions formed in the most extensively purified protein fraction displayed deletions containing short patches of sequence homology at their break points, a feature characteristic of single-strand annealing (SSA). Detailed biochemical characterisation revealed the presence of DNA ligase III, DNA polymerase epsilon, FEN-1 endonuclease, and exonuclease activities of 5'-3' and 3'-5' directionality. We show that these activities are able to correctly process proposed intermediates of SSA. Interestingly, neither Ku nor the associated DNA-dependent protein kinase were detected, indicating that the mechanism can dispense with Ku. Our findings provide evidence for the existence of an error-prone NHEJ pathway that creates deletions by microhomology driven SSA. PMID- 9874204 TI - A peptide isolated from a random phage peptide library is a structural mimic to the P1, P4-diadenosine 5'-tetraphosphate binding site on its receptor. AB - We have previously demonstrated that a monoclonal antibody (mAb TL4), which inhibits P1, P4-diadenosine 5'-tetraphosphate (Ap4A) binding to its receptor, selected a consensus RGS tripeptide from a random phage hexapeptide library. RGS interfered with Ap4A binding to its membrane receptor [Liu, G., Bryant, R. T. & Hilderman, R. H. (1996) Biochemistry 35, 197-201]. However the mechanism by which RGS interfered with Ap4A binding to its receptor was not determined. In this communication, we demonstrate that RGS interacts with Ap4A to prevent [3H]Ap4A binding to the Ap4A membrane receptor. To further characterize the mechanism by which RGS inhibits Ap4A binding to its receptor, we used mAb TL4 to screen a 15 residue random peptide phage library and DNAs from 24 clones were sequenced. 20 clones contain a RGSSS sequence while 17 of these clones contained an identical 15-amino-acid insert (clone A). Gel-filtration studies of previously equilibrated clone A phage and [3H]Ap4A support the idea that [3H]Ap4A interacts specifically with clone A phage while RGS effectively competes for [3H]Ap4A interaction on clone A phage. These data are consistent with RGS mimicking a sequence on the receptor essential for Ap4A binding. PMID- 9874205 TI - Isolation of a cDNA from Arabidopsis thaliana that complements the sec14 mutant of yeast. AB - The SEC14 gene of Saccharomyces cerevisiae codes for a phosphatidylinositol transfer protein (Sec14p(sc)) which is capable of transferring both phosphatidylinositol and phosphatidylcholine between membranes in vitro. Genetic and biochemical studies conducted in S. cerevisiae have shown that this protein acts as an inhibitor of phosphatidylcholine biosynthesis via the so-called Kennedy pathway only. This inhibition is controlled by the binding of phospholipids to the Sec14p(sc) protein. Here we describe the isolation of a cDNA from Arabidopsis thaliana by functional complementation of a sec14(ts) mutant of S. cerevisiae. This cDNA, designated AtSEC14, is capable of restoring the growth of the sec14(ts) mutant at the restrictive temperature of 37 degrees C. Extracellular invertase measurements indicated that the cDNA can partly restore protein secretion. In addition, the phosphatidylinositol-transfer activity measured in protein extracts is greatly enhanced in the complemented mutant strain when compared with the sec14(ts) mutant. The best sequence similarity at the amino acid level is found with the Sec14p protein of S. cerevisiae (36.5% similarity), and most of the amino acids that are thought to be involved in the binding of phospholipids in the yeast protein are conserved in the AtSEC14 gene product. Southern analysis suggests the presence of a single gene in the Arabidopsis genome, although the existence of distantly related sequences cannot be excluded. This gene is expressed in roots, leaves, flowers and siliques of Arabidopsis. PMID- 9874206 TI - Refolding of native and recombinant chicken riboflavin carrier (or binding) protein: evidence for the formation of non-native intermediates during the generation of active protein. AB - Riboflavin carrier (or binding) protein (RCP) is a phosphoglycoprotein originally purified from the egg white, yolk and serum of laying hens. The 18 cysteine residues present in RCP form nine disulfide bridges, allowing the protein to form a compact structure to generate a hydrophobic pocket in which riboflavin sits. We studied the refolding of totally reduced and denatured egg white RCP and found that the protein initially folded to generate a molecule that did not possess riboflavin-binding activity, despite near-complete oxidation of the cysteine residues. Riboflavin-binding activity was then slowly regained, but the final refolded form of the protein was less compact in structure than the native molecule, due to incomplete oxidation of all the cysteine residues. Denatured and reduced dephosphorylated RCP refolded as efficiently as the native protein, with similar rates of disulfide-bond oxidation and generation of riboflavin binding, showing that the phosphoserine stretch of RCP has little role to play during refolding. In order to study the role of glycosylation in the refolding process, the cDNA for full-length RCP was expressed in Escherichia coli and purified. Recombinant RCP refolded only in the presence of redox buffers, demonstrating that glycosylation of RCP could allow the formation of high yields of productive intermediates in the folding pathway. Using a panel of conformation-specific monoclonal antibodies to RCP, it appeared that the folding intermediates of RCP possessed a structure distinctly different to the native protein, indicating that the correct folding pathway of RCP passed through conformation(s) generated by non-native disulfide bridges. PMID- 9874208 TI - Sporulation-specific expression of the yeast DIT1/DIT2 promoter is controlled by a newly identified repressor element and the short form of Rim101p. AB - Expression of the yeast genes DIT1 and DIT2 is confined to mid/late sporulation. Transcription of these two divergently arranged genes is controlled by a common 900-bp intergenic region. Random mutagenesis of this promoter and tests with appropriate reporter constructs identified an 11-bp cis-acting palindromic sequence, DIT repressor element (DRE), as a major negative regulatory site during vegetative growth. Repression is exerted by DRE in conjunction with a mid sporulation element (MSE)-like sequence situated 26 bp away. These sequence elements are both contained within the 76-bp negative regulatory element (NRE) defined previously [Friesen H., Hepworth, S. R. & Segall, J. (1997) Mol. Cell. Biol. 17, 123-134]. The activated form of Rim101p, a transcriptional inducer of the early meiotic gene IME1, enhances expression from the DIT1 promoter both in vegetative and sporulating cells. Activation by Rim101p does not seem to involve binding of Rim101p at either of the two cis-acting sites described here, since reporter constructs with both elements or most of the NRE deleted could still be activated by the short form of Rim101p. PMID- 9874207 TI - cDNA cloning, in vitro expression, and biochemical characterization of cholinesterase 1 and cholinesterase 2 from amphioxus--comparison with cholinesterase 1 and cholinesterase 2 produced in vivo. AB - We have isolated cDNAs coding for the complete amino acid sequences of cholinesterase 1 (ChE1) and cholinesterase 2 (ChE2) from amphioxus. Both ChE transcripts have the characteristics of H-type catalytic subunits, which are inserted in the membrane via an ethanolamine-glycan-phosphatidylinositol anchor. The members of the catalytic triad of ChEs, the three pairs of cysteine residues involved in intrachain disulfide bonding, a cysteine near the carboxy terminal of both sequences, which could mediate interchain disulfide bonding, and 11 of the 14 aromatic amino acids that line the catalytic gorge of AChE are conserved. A remarkable difference between the two enzymes is in the region of the acyl binding pocket, which plays an important role in determining substrate specificity in cholinesterases. ChE2 contains a sequence that resembles the acyl pocket of invertebrate ChE, while the acyl-binding site of ChE1 is novel. There are also differences between the two enzymes in the peripheral anionic site, which mediates inhibition by certain ligands. In vitro expression in COS-7 cells demonstrates that ChE2 hydrolyzes acetylthiocholine almost exclusively, while ChE1 hydrolyzes both acetylthiocholine and butyrylthiocholine. Both enzymes are inhibited comparably by BW284c51, but ChE1 is considerably more resistant to inhibition by propidium, ethopropazine, and eserine than is ChE2. Velocity sedimentation indicates that ChE1 and ChE2 are present as amphiphilic and nonamphiphilic G2 forms in vivo and in vitro. Another molecular form, which sediments at 17 S, is also present in vivo. Nondenaturing gel electrophoresis in conjunction with digestion by phosphatidylinositol-specific phospholipase C demonstrates that the vast majority of ChE1 and ChE2 is present as ethanolamine glycan-phosphatidylinositol-anchored G2 forms in vivo. ChE1 also possesses an ethanolamine-glycan-phosphatidylinositol-anchor in vitro; however, ChE2 produced in vitro could not be detected on nondenaturing gels. PMID- 9874209 TI - Structure/function correlation of spermine-analogue-induced modulation of peptidyltransferase activity. AB - In a cell-free system derived from Escherichia coli, various analogues of spermine were used to study their effect on the binding of AcPhe-tRNA to poly (U) programmed ribosomes and on the puromycin reaction carried out at 6 mM Mg2+ (Ac, acetyl). In the absence of factors washable from ribosomes (FWR fraction), mono acylated or di-acylated analogues of spermine stimulate the binding of AcPhe-tRNA to a lesser degree than spermine, in the order: N1-acetylspermine > N1,N12 diacetylspermine approximately = N1,N12-dipivaloylspermine. Also, the above analogues do not show any sparing effect on Mg2+ requirements for AcPhe-tRNA binding to ribosomes, in contrast to spermine. The presence of FWR fraction during the binding or acetylation of the secondary amines of spermine moderates or abolishes the stimulatory effect. In addition, all analogues tested enhance the stability of the ternary complex AcPhe-tRNA-poly(U)-ribosome and the extent of AcPhe-puromycin synthesis, particularly in the absence of the FWR fraction. At the kinetic phase of AcPhe-puromycin synthesis, the analogues display both stimulatory and inhibitory effects, depending on the absence (partial noncompetitive inhibition) or the presence of the FWR fraction (nonessential activation in concert with partial noncompetitive inhibition). Detailed kinetic analysis shows that the analogues tested can mimic the behaviour of spermine, however, the potency to affect the peptidyltransferase activity depends on their degree of acylation, acyl-substituent size, charge distribution and on their chain flexibility. PMID- 9874210 TI - Disulfide bond mapping and structural characterization of spruce budworm antifreeze protein. AB - The 9-kDa, Thr-, Ser-, and Cys-rich thermal hysteresis protein from spruce budworm (sbwTHP) is 10-30 times more effective than fish antifreeze proteins (AFPs) at depressing solution freezing points via ice-crystal growth inhibition. Since this insect protein is only available in microgram quantities from its natural source, recombinant sbwTHP was produced from inclusion bodies in Escherichia coli by a refolding protocol. Incompletely folded forms were removed during ion-exchange and reverse-phase chromatography, resulting in fully active sbwTHP that was indistinguishable in its properties from native sbwTHP. The antifreeze was completely inactivated by reduction, showed no reaction with sulfhydryl reagents, and was not inhibited by EDTA. All eight cysteine residues appear to be involved in disulfide bond formation. Tryptic cleavage and peptide analysis is consistent with linkages between the first and second cysteine residues, the third and fourth, fifth and eighth, and the sixth and seventh. NMR analysis confirmed that the fully folded form of sbwTHP was well structured and had a single conformation. Both NMR and CD spectra indicate the presence of extensive beta structure (70-80%) with little or no alpha helix. The protein maintains antifreeze activity over a broad range of pH values, and its conformation is independent of both temperature (over the range 0 degrees C to 20 degrees C), and the presence of 50% trifluoroethanol. PMID- 9874211 TI - Molecular cloning and expression of active Ole e 3, a major allergen from olive tree pollen and member of a novel family of Ca2+-binding proteins (polcalcins) involved in allergy. AB - A cDNA encoding Ole e 3, a major allergen from olive-tree pollen, has been cloned and sequenced. A strategy based on two-step PCR amplification towards the 5' end and 3' end, with an internal specific primer, has been used. The isolated cDNA contains an open reading frame coding for a polypeptide of 84 amino acids, which is in agreement with the composition and molecular mass of the natural allergen, exhibiting two 12-residue segments homologous to Ca2+-binding sites of EF-hand type. The cDNA was inserted into the pET-11b expression vector and over-expressed in Escherichia coli. The purified recombinant protein shows identical secondary structure to that of the natural allergen and is able to bind both IgE from sera of patients allergic to olive pollen and polyclonal antibodies raised against olive-pollen Ole e 3. The capacity of binding Ca2+ has been demonstrated for both natural and recombinant allergens. RNA transcripts of Ole e 3 were only detected in pollen tissue. Northern-blot and Western-blot analyses of poly(A)+ RNA and protein extracts, respectively, obtained from a variety of olive-tree-related and nonrelated mature pollens demonstrated the presence of Ole e 3 homologous proteins. This indicates a sequence conservation and widespread distribution for this family of Ca2+-binding proteins that can be responsible for allergenic cross reactivity. We suggest the tentative generic name of polcalcins for the members of this family of Ca2+-binding proteins from pollen. PMID- 9874212 TI - The amino acid sequence of the ram spermatidal protein 3--a transition protein TP3 or TP4? AB - As in other mammals, several nuclear basic proteins replace histones during the differentiation of germinal cells into spermatozoa in the ram. These proteins called transition proteins (TP) are later replaced by protamines. The amino acid sequence of the ram spermatidal protein 3 has been established by Edman degradation of the protein and of its fragments generated from digestion with endoproteinase Lys-C and pepsin and from the coding sequence of the gene and of the cDNA. The ram protein 3 is a basic protein of 109 residues (calculated Mr 13,200) with arginine and lysine residues uniformly distributed along the polypeptide chain. Of the 13 serine and threonine residues, 9 are located in structural motifs where they could be phosphorylated and, thus, modulate the binding of the protein to DNA. The tyrosine residues at position 33 and position 93, located in a basic environment, and the tryptophan residue at position 29 could be involved in the interactions of the protein with DNA through the stacking of their aromatic ring between the nucleotide bases. The ram protein 3 differs completely from the two well-defined transition protein families TP1 and TP2, which are also synthesised transiently during mammal spermiogenesis. In contrast with the rat TP3, ram protein 3 does not correspond to a precursor of a protamine. However, it shares structural similarities with both transition proteins TP3 and TP4 of the boar. The ram protein 3 and the boar transition proteins TP3 and TP4 probably belong to the same transition-protein group and would play similar functions in the chromatin remodelling during spermiogenesis. As protamine and transition-protein TP1 and TP2 genes from mammals, the coding sequence of the gene of ram protein 3 is interrupted by one intron but its organisation is different. PMID- 9874213 TI - The solution structure of parsley [2Fe-2S]ferredoxin. AB - The [2Fe-2S]ferredoxin I (Fd I) from parsley leaves (Mr = 10,500; 96 amino acids) in the Fe(III)-Fe(III) oxidized form has been studied by 1H-NMR spectroscopy. Sequence-specific 1H-NMR assignments were obtained through two-dimensional classical double-quantum-filtered-COSY, NOESY and TOCSY spectra. NOEs between protons as close as 5.6 A from the paramagnetic Fe(III) atoms were observed at 800 MHz. A total of 3066 NOEs (of which 2533 are meaningful) and 18 distance constraints taken from X-ray crystallography of the Fe2S2 active site were used to obtain the solution structure. From inversion recovery NOESY experiments, 33 longitudinal relaxation rate (Qpara) constraints were used for the structural refinement. The final structure was obtained by a process of restrained energy minimization. Root-mean-square (rmsd) deviation values obtained for the family of 18 structures (with reference to the average structure) are 0.52 +/- 0.10 A and 0.91 +/- 0.12 A for backbone and all heavy atoms respectively. The structure consists of seven-strands of beta-sheets and four short alpha-helices. The quality of the present solution structure is among the best of those reported for [2Fe-2S]ferredoxins. The secondary structure and overall folding are compared with those of Anabaena variabilis Fd and the higher plant Equistum arvense (horse tail) protein determined through X-ray crystallography. The groups believed to be responsible for electron transfer have been analysed. PMID- 9874214 TI - Membrane topology of the myelin/oligodendrocyte glycoprotein. AB - Myelin/oligodendrocyte glycoprotein (MOG), a specific component of the mammalian central nervous system, is located on the surface of the oligodendrocyte plasma membrane and the outermost lamellae of mature myelin; it is expressed during the latter steps of myelinogenesis. It has been shown that MOG may play a pathological role in autoimmune demyelinating diseases of the central nervous system, although its physiological function remains unknown. MOG is an integral membrane glycoprotein with an extracellular immunoglobulin-like domain and two hydrophobic segments which were predicted to be membrane-spanning on the basis of hydropathy analysis. As a first step in elucidation of MOG function, we have investigated its membrane topology, combining immunofluorescence studies on cultured oligodendrocytes and MOG-transfected Chinese hamster ovary cells with biochemical analyses, including in vitro translation, membrane insertion and protease-digestion assays. Our results indicate that the C-terminal tail of MOG is located into the cytoplasm, and that only the first hydrophobic region of MOG spans the membrane whereas the second hydrophobic region appears to be semi embedded in the lipid bilayer, lying partially buried in the membrane with its N terminal and C-terminal boundaries facing the cytoplasm. PMID- 9874216 TI - Expression of genes encoding the E2 and E3 components of the Bacillus stearothermophilus pyruvate dehydrogenase complex and the stoichiometry of subunit interaction in assembly in vitro. AB - Genes encoding the dihydrolipoyl acetyltransferase (E2) and dihydrolipoyl dehydrogenase (E3) components of the pyruvate dehydrogenase (PDH) multienzyme complex from Bacillus stearothermophilus were overexpressed in Escherichia coli. The E2 component was purified as a large soluble aggregate (molecular mass > 1 x 10(6) Da) with the characteristic 532 symmetry of an icosahedral (60-mer) structure, and the E3 as a homodimer with a molecular mass of 110 kDa. The recombinant E2 component in vitro was capable of binding either 60 E3(alpha2) dimers or 60 heterotetramers (alpha2beta2) of the pyruvate decarboxylase (E1) component (also the product of B. stearothermophilus genes overexpressed in E. coli). Assembling the E2 polypeptide chain into the icosahedral E2 core did not impose any restriction on the binding of E1 or E3 to the peripheral subunit binding domain in each E2 chain. This has important consequences for the stoichiometry of the assembled complex in vivo. The lipoyl domain of the recombinant E2 protein was found to be unlipoylated, but it could be correctly post-translationally modified in vitro using a recombinant lipoate protein ligase from E. coli. The lipoylated E2 component was able to bind recombinant E1 and E3 components in vitro to generate a PDH complex with a catalytic activity comparable with that of the wild-type enzyme. Reversible unfolding of the recombinant E2 and E3 components in 6 M guanidine hydrochloride was possible in the absence of chaperonins, with recoveries of enzymic activities of 95% and 85%, respectively. However, only 26% of the E1 enzyme activity was recovered under the same conditions as a result of irreversible denaturation of both E1alpha and E1beta. This represents the first complete post-translational modification and assembly of a fully active PDH complex from recombinant proteins in vitro. PMID- 9874215 TI - Reactivity studies of the tyrosyl radical in ribonucleotide reductase from Mycobacterium tuberculosis and Arabidopsis thaliana--comparison with Escherichia coli and mouse. AB - Ribonucleotide reductase (RNR) is a key enzyme for DNA synthesis since it provides cells with deoxyribonucleotides, the DNA precursors. Class I alpha2beta2 RNRs contain a dinuclear iron center and an essential tyrosyl radical in the beta2 component (protein R2). This is also true for the purified protein R2 of Mycobacterium tuberculosis RNR, as shown by iron analysis, light absorption and EPR spectroscopy. EPR spectroscopy at 286 GHz revealed a high g(x) value, suggesting that the radical is not hydrogen bonded, as in other prokaryotic R2s and in contrast with eukaryotic R2s (from Arabidopsis thaliana and mouse). Furthermore, it proved to be very resistant to scavenging by a variety of phenols and thiols and by hydroxyurea, similar to the Escherichia coli radical. By comparison, the plant and mouse radicals are very sensitive to drugs such as resveratrol and 2-thiophenthiol. The radical from M. tuberculosis RNR does not seem to be an appropriate target for new antituberculous agents. PMID- 9874217 TI - Solution structure of an artificial Fe8S8 ferredoxin: the D13C variant of Bacillus schlegelii Fe7S8 ferredoxin. AB - The solution structure of the D13C variant of the thermostable Fe7S8 ferredoxin from Bacillus schlegelii has been determined by 1H-NMR spectroscopy in its oxidized form. In a variable-temperature NMR study the D13C variant was as thermostable (up to 90 degrees C) as the wild-type protein (WT). Seventy-five out of 77 amino acid residues and 81% of all theoretically expected proton resonances in the D13C Fe8S8 protein have been assigned. Its structure was determined through torsion angle dynamics calculations with the program DYANA, using 935 meaningful NOEs (from a total of 1251), hydrogen bond constraints, and NMR derived dihedral angle constraints for the cluster-ligating cysteines. Afterwards, restrained energy minimization and restrained molecular dynamics were applied to each conformer of the family. The final family of 20 structures has RMSD values from the mean structure of 0.055 nm for the backbone atoms and of 0.099 nm for all heavy atoms. The overall folding of the WT is maintained in the mutant, except for the immediate vicinity of the new cysteine, which becomes much more similar to native Fe8S8 proteins. The two residues at positions 11 and 12, which constitute an insertion with respect to all known Fe8S8 proteins, assume a conformation that does not prevent the preceding and following residues from folding like in native Fe8S8 proteins. Clear evidence for the existence of two conformations involving almost half of the amino acid residues was found. The two conformations are structurally indistinguishable. Temperature-dependent NMR experiments show that one of them is thermodynamically more stable than the other. PMID- 9874218 TI - Analysis of the clumping-mediating domain(s) of sex pheromone plasmid pAD1 encoded aggregation substance. AB - Aggregation substance, a cell surface adhesin encoded on sex-pheromone plasmids exclusively found in the opportunistic pathogen Enterococcus faecalis, displays a dual function in that it mediates (a) adhesion to host tissues and (b) aggregation of E. faecalis cells to result in efficient plasmid transfer. While there have been many investigations regarding the regulation of inducible plasmid transfer and involvement in pathogenicity, nearly nothing is known about the structural basis of clumping capacity intrinsic to aggregation substance. Here, data are presented regarding the respective domain(s) of Asa1 (the adhesin encoded on plasmid pAD1) by analyzing the effects of in-frame deletions on clumping and by measuring binding of E. faecalis cells to surface-bound subfragments of aggregation substance. These data indicate that the N-terminal part of the adhesin is responsible for clumping, with a region between amino acid 525 and amino acid 617 playing a dominant role. Furthermore, by raising antibodies against different fragments of Asa1, it is shown that aggregation substance contains cross-reacting epitopes in its C-terminal and N-terminal parts, which could not be derived from their primary sequences. PMID- 9874219 TI - Solution structure of the recombinant oxidized rabbit uteroglobin using homonuclear and heteronuclear multidimensional NMR. AB - Rabbit uteroglobin (rab-UG) is a 16-kDa homodimeric secretory protein with potent anti-inflammatory/immunomodulatory properties. Its physiological role is still unclear, although it was observed that several small hydrophobic molecules bind to the oxidized and the reduced uteroglobin. It is suggested that the formation and/or disruption of the two disulphide bridges not only regulates this binding itself, but also the affinity to the ligand. The determination of the solution structure has been started with the assignment of 1H, 15N and 13C resonances of the oxidized rabbit uteroglobin, based on several two-dimensional and three dimensional homonuclear and heteronuclear double and triple resonance experiments. The assignment was possible with the overproduction of the wild-type as well as of uniformly 15N-labeled and 15N/13C-labeled samples of the recombinant protein. A complete assignment of 1H, 15N and 13C resonances, the secondary-structure elements and the tertiary structure in solution is presented. The tertiary solution structure was found to be in good agreement with the previously determined crystal structure of rab-UG and with the solution structure of human uteroglobin (h-UG). h-UG and rab-UG are extremely stable proteins within a wide range of pH and temperatures. Some of the binding characteristics of ligands of rab-UG and a mutant with all cysteine residues exchanged to serine residues are discussed. PMID- 9874220 TI - The secondary structure and architecture of human elastin. AB - The presented work constitutes the first structural characterization of both insoluble human elastin and its solubilized form, kappa-elastin. Structural data were reached following the use of Fourier transform infrared, near infrared Fourier transform Raman and circular dichroism optical spectroscopic methods and their quantitative analysis permitted us to estimate approximately 10% alpha helices, approximately 35% beta-strands and approximately 55% undefined conformations in the global secondary structure of insoluble human elastin in the solid state. Following the use of the LINK method, the probable local distribution of the secondary-structure elements along the sequence was determined and compared to that obtained for bovine elastin, the historical standard of elastin. This comparison led us to propose a globular architecture for the human elastomer and permitted us to delineate some elements of its structure-elasticity relationship. PMID- 9874221 TI - The deoxyribonuclease activity attributed to ribosome-inactivating proteins is due to contamination. AB - The mode of action of ribosome-inactivating proteins (RIPs) has, for many years, been considered to be depurination of a specific adenyl residue of ribosomal RNA, resulting in inhibition of protein synthesis. Recently, this view has been challenged by the observation that many RIP preparations have significant DNase activity in addition to their N-glycosidase activity. In this study, we have investigated the putative DNase activity of two RIPs, ricin and pokeweed antiviral protein (PAP), and show that, in both cases, the DNase activity is due to the presence of contaminating nucleases. The N-glycosidase and DNase activities of PAP were separately and specifically inactivated by chemical modification and heat. Gel filtration of ricin allowed physical separation of the two activities. Furthermore, neither recombinant PAP nor recombinant ricin A chain purified from Escherichia coli displayed DNase activity. PMID- 9874222 TI - Does an asparaginyl-specific cysteine endopeptidase trigger phaseolin degradation in cotyledons of kidney bean seedlings? AB - An asparaginyl-specific cysteine endopeptidase which was named 'legumain-like proteinase' (LLP) and has an apparent molecular mass of 38.1 kDa was isolated from cotyledons of kidney bean (Phaseolus vulgaris L.) seedlings and partially characterized. It is, to our knowledge, the first known proteinase which in vitro extensively degrades native phaseolin, the major storage globulin of this grain legume. Phaseolin that in vitro had been partially degraded by LLP (Pvitro) and phaseolin that was isolated after partial in vivo breakdown 6 days after the start of seed imbibition (Pvivo) showed similar fragment patterns on SDS/polyacrylamide gels. The fragments had identical cleavage sites in Pvitro and Pvivo as determined by partial amino acid sequencing. In both types of partially degraded phaseolin, these cleavage sites have asparagine in the P1 position. Two of the cleavage sites are located in the beta-barrel domain of the C-terminal module and only one cleavage site was found in the beta-barrel domain of the N terminal module according to the consensus structural model of phaseolin subunits. These results suggest that very likely LLP could in vivo be responsible for the initiation of phaseolin proteolysis. Two different legumain-specific clones named cp6b and p21b were isolated from a cDNA library of germinated bean cotyledons. Cp6b encodes LLP, while p21b encodes a VPE-like enzyme. Southern-blot analysis revealed a single gene copy for Pv-VPE and, presumably, at least two gene copies for LLP in the kidney bean genome. Northern-blot analysis indicated that mRNAs for both clones appear de novo during seed germination. However, the developmental patterns of the transcript levels corresponding to the two clones differed significantly. The temporal pattern of phaseolin degradation and of LLP polypeptide levels agreed well with the suggestion that LLP plays a key role in the mobilization of phaseolin during and after kidney bean germination. PMID- 9874223 TI - The surface-charge asymmetry and dimerisation of cytochrome c550 from Paracoccus denitrificans--implications for the interaction with cytochrome c peroxidase. AB - The implications of the dimeric state of cytochrome c550 for its binding to Paracoccus cytochrome c peroxidase and its delivery of the two electrons required to restore the active enzyme during catalysis have been investigated. The amino acid sequence of cytochrome c550 of Paracoccus denitrificans strain LMD 52.44 was determined and showed 21 differences from that of strain LMD 22.21. Based on the X-ray structure of the latter, a structure for the cytochrome c550 monomer from strain 52.44 is proposed and a dipole moment of 945 debye was calculated with an orientation close to the exposed haem edge. The behaviour of the cytochrome on molecular-exclusion chromatography is indicative of an ionic strength-dependent monomer (15 kDa)/dimer (30 kDa) equilibrium that can also be detected by 1H-NMR spectroscopy. The apparent mass of 50 kDa observed at very low ionic strength was consistent with the presence of a strongly asymmetric dimer. This was confirmed by cross-linking studies, which showed that a cross-linked species of mass 30 kDa on SDS behaved with an apparent mass of 50 kDa on molecular-exclusion chromatography. A programme which carried out and evaluated molecular docking of two monomers to give a dimer generated a most probable dimer in which the monomer dipoles lay almost antiparallel to each other. The resultant dipole moment of the dimer is therefore small. Although this finding calls into question the possibility of preorientation of a strongly asymmetrically charged cytochrome as it collides with a redox partner, the stoichiometry of complex formation with cytochrome c peroxidase as studied by 1H-NMR spectroscopy shows that it is the monomer that binds. PMID- 9874224 TI - Characterization of a calcium-dependent calmodulin-binding domain in the 135-kD human protein 4.1 isoform. AB - The putative calmodulin binding domain of non-erythroid protein 4.1, previously suggested by Kelly et al. [Kelly, G. M., Zelus, B. D. & Moon, R. T. (1991) J. Biol. Chem. 266, 12469-12473] has been synthesized, and its binding to calmodulin has been studied by fluorescence spectroscopy. For this purpose, the peptide has been N-terminally dansylated. The 4.1 peptide Dns-Abu-S76RGLSRLFSSFLKRPKS92, binds calmodulin in a calcium-dependent way with high affinity (Kd = 23 +/- 6 nM). The peptide inhibits bovine-heart phosphodiesterase with an IC50 of 50 nM. Since the sequence of the peptide shows two putative consensus sites of phosphorylation by cAMP-dependent protein kinase or Ca2+-calmodulin protein kinase II, the interaction of the two mono-phosphorylated peptides (P4.1 Ser(80 P) and P4.1 Ser(84-P)) and the di-phosphorylated peptide (P4.1 Ser(80-P)/Ser(84 P)) with calmodulin has been investigated. A decrease of affinity by a factor 1.5 8 has been observed for the phosphorylated peptides. CD measurements have shown an increase of the content of alpha helices in the peptides when bound to calmodulin. PMID- 9874225 TI - A low-spin iron with CN and CO as intrinsic ligands forms the core of the active site in [Fe]-hydrogenases. AB - In this report the first high-quality infrared spectra of [Fe]-hydrogenase are presented. Analyses of these spectra obtained under a variety of redox conditions strongly indicate that [Fe]-hydrogenases contain a low-spin Fe ion in the active site with one CN- group and one CO molecule as intrinsic, non-protein ligands. When in the ferric state, the presence of such an ion can explain the enigmatic EPR properties (the rhombic 2.10 signal) of the active, oxidised enzyme. To account for other, well-characterised properties of the active site, we propose that the active site of [Fe]-hydrogenases consists of this low-spin Fe ion bound to an unusual [4Fe-4S] cluster via bridges with sulphur atoms. PMID- 9874226 TI - Inactivation of isocitrate dehydrogenase kinase/phosphatase by 5'-[p (fluorosulfonyl)benzoyl]adenosine is not due to the labeling of the invariant lysine residue found in the protein kinase family. AB - The ATPase activity of Escherichia coli isocitrate dehydrogenase kinase/phosphatase was rapidly lost after prior incubation with the ATP analogue 5'-[p-(fluorosulfonyl)benzoyl]adenosine (FSBA). This inactivation was prevented by the presence of either 5 mM ATP or 5 mM ADP plus Mg2+, while it could be fully reversed by subsequent addition of dithiothreitol, thereby indicating the involvement of cysteine residue(s) in this process. About 2 mol [3H]FSBA/mol IDHK/P were bound during the time course of the inactivation. However, this binding was not significantly modified by either prior incubation with ATP or subsequent addition of dithiothreitol. This suggested that FSBA-mediated inactivation of isocitrate dehydrogenase kinase/phosphatase occurred via the formation of a disulfide bond. Accordingly, mass spectral analysis revealed that on addition of FSBA, a disulfide bond was formed between residues Cys356 and Cys523. The mutation Cys356Ser renders the enzyme insensitive to FSBA treatment indicating that Cys356 is the primary target for this analogue. However, the Cys523Ser mutant was still inactivated by FSBA and mass spectral analysis showed that this was due to the formation of a new disulfide bond between Cys356 and Cys480. PMID- 9874227 TI - Purification and properties of the chloroplastic form of biotin holocarboxylase synthetase from Arabidopsis thaliana overexpressed in Escherichia coli. AB - Holocarboxylase synthetases (HCSs) are key enzymes in biotin utilisation in both prokaryotes and eukaryotes. In a previous study, we demonstrated that, in plants, HCS activity is localised in cytosol, chloroplasts and mitochondria. We also described the cloning and sequencing of a full-length cDNA encoding an Arabidopsis thaliana HCS isoform with a putative organelle-transit peptide. In the study reported here, this cDNA was used to construct an overproducing Escherichia coli strain. The recombinant enzyme was isolated using an efficient three-step purification procedure. Polyclonal antibodies raised against pure HCS were produced to elucidate the subcellular localisation of this protein. Immunodetection carried out by Western blotting of isolated pea leaf subcellular compartments specifically revealed a single polypeptide that was ascribed to the chloroplast compartment. Immunocytochemistry of thin-cut sections from tobacco leaves, transformed by the complete coding sequence of A. thaliana HCS cDNA via Agrobacterium tumefaciens, confirmed that the enzyme encoded by this cDNA is the chloroplastic isoform. Moreover, physicochemical, biochemical and kinetic properties of the pure recombinant HCS were determined. The native recombinant enzyme is a 37-kDa monomer. In contrast to the major part of HCS activity measured in leaf extracts, the recombinant chloroplastic enzyme did not require addition of Mg2+ to be fully active, but was substantially inhibited by EDTA. This suggested that the chloroplastic HCS may contain a tightly-bound divalent cation required for enzyme activity. The recombinant enzyme was able to biotinylate efficiently apo-biotin carboxyl carrier protein (BCCP) from E. coli and apo-methylcrotonoyl-CoA carboxylase (MCCase) from A. thaliana. Apparent Km values for the enzyme substrates D-biotin, ATP and apo-MCCase were found to be 130 nM, 4.4 microM and 32 microM, respectively. Steady-state kinetic analyses of the HCS-catalysed reaction were investigated with respect to reaction mechanism and inhibition by AMP, one of the end-products of the enzyme-catalysed reaction. Substrate interaction and product inhibition patterns indicated that ATP and D biotin bind sequentially, in an ordered manner, to the enzyme and that ATP or D biotin and apo-BCCP bind in ping-pong fashion. PMID- 9874228 TI - Activation and reaction kinetics of the dimethylamine/coenzyme M methyltransfer in Methanosarcina barkeri strain Fusaro. AB - Dimethylamine/5-hydroxybenzimidazolylcobamide methyltransferase (DMA-MT) from Methanosarcina barkeri Fusaro catalyzes (Vmax = 4700 nmol x min(-1) x mg(-1) protein; k(cat) = 7.8 s(-1)) the transfer of a methyl group from dimethylamine (apparent Km = 0.45 mM) to its corrinoid prosthetic group to yield monomethylamine (MMA) and the methylated enzyme. The product, MMA, is a competitive inhibitor of the reaction (apparent Ki = 5.5 mM). The methyl group bound to the corrinoid prosthetic group of DMA-MT is subsequently transferred to coenzyme M in a reaction mediated by methylcobalamin/coenzyme M methyltransferase isoenzyme II [MT2(II)], which binds with high affinity to DMA-MT (apparent Km = 0.22 microM). As isolated, DMA-MT is inactive, but it can enzymically be reactivated by methyltransferase activating protein (MAP), ATP, and hydrogenase. Apart from the established role in corrinoid activation, ATP was found to act as a powerful allosteric effector on the methyltransferase reaction. The results of kinetic studies, supported by the resolution of as-yet partially purified auxiliary protein fractions, demonstrate that DMA-MT, MT2(II), MAP, and hydrogenase are the only enzymic components involved in the dimethylamine/coenzyme M methyltransfer in M. barkeri Fusaro. PMID- 9874230 TI - Identification of highly fucosylated N-linked oligosaccharides from the human parotid gland. AB - The glycosylation of a number of constituents of human saliva is known to modify its biological roles, such as its lubricating properties and binding of microbial flora. Gillece-Castro et al. [Gillece-Castro, B. L., Prakobphol, A., Burlingame, A. L., Leffler, H. & Fisher, S. J. (1991) J. Biol. Chem. 266, 17358-17368] have proposed that the major glycan on the salivary proline-rich glycoproteins is a trifucosylated biantennary sugar with one difucosylated and one unfucosylated antenna. Furthermore, they proposed that the non-fucosylated antenna mediated adherence to a peridontal pathogen, Fusobacterium nucleatum. The detailed structures and roles of other highly fucosylated glycans that co-exist in the parotid gland are not fully known. In view of the influence of outer-arm fucosylation on carbohydrate recognition processes in general, this paper reports the use of a combination of HPLC (normal and reversed phase), matrix-assisted laser-desorption/ionisation (MALDI) mass spectrometry and exoglycosidase digestions to dissect the detailed structures of the most abundant of these polyfucosylated glycans. For measurement of reversed-phase HPLC retention times, new calibration units were used which paralleled the glucose units used for normal-phase HPLC. These differed in that the difference in retention times were compared with those derived from a ladder of 2-aminobenzamide-labelled arabinose oligomers instead of the corresponding oligomers from partially hydrolysed dextran. Over sixty neutral sugars were identified from the parotid gland and many of these were additionally found substituted with sialic acid (both alpha2-3 linked and alpha2-6-linked) and sulphate. These glycans were mainly bi- and tri antennary sugars with up to five and seven fucose residues respectively, containing fucose alpha1-3-linked to the outer-arm GlcNAc residues and alpha1-2 linked to the galactose. All fucosylated structures contained a core (alpha1-6 linked) fucose. The detailed structure of the trifucosylated biantennary glycan was confirmed, together with the structures of another 12 fucosylated biantennary glycans. Smaller amounts of hybrid and tetraantennary structures were also found and bisected glycans were shown to be constituents of parotid glycoproteins for the first time. Acidic glycans were mainly substituted with sialic acid. Most were monosialylated as the presence of fucose on the antennae was found to suppress the addition of extra sialic acid moieties. The possible functional significance of highly fucosylated N-glycans is discussed in relation to their modification of the availability of other non-reducing terminal monosaccharides for recognition processes. PMID- 9874231 TI - Structure of the O polysaccharide and immunochemical relationships between the lipopolysaccharides of Pseudomonas syringae pathovar tomato and pathovar maculicola. AB - O polysaccharides (OPS) of the lipopolysaccharides (LPS) of Pseudomonas syringae pathovar tomato GSPB 483 and pathovar maculicola IMV 381 were studied by 1H-NMR and 13C-NMR spectroscopy, including two-dimensional COSY, rotating-frame NOE spectroscopy (ROESY), and H-detected 1H,13C heteronuclear multiple-quantum coherence (HMQC) experiments. The OPS from both strains were shown to be chemically identical. Two structurally different types of repeating units (O repeats 1 and 2) were elucidated in each OPS. The minor O repeat is a pentasaccharide having the structure 2. The same structure has been reported earlier for some other OPS of P. syringae. The major O repeat is a hexasaccharide with the novel structure 1 which is distinguished from 2 by the presence of the second lateral residue of 3-acetamido-3,6-dideoxy-D-galactose (D-Fuc3NAc) and by a different position of substitution of one of the L-rhamnose (Rha) residues in the backbone. [structures: see text] Structural and serological diversity of OPS of strains belonging to P. syringae pv. tomato and pv. maculicola and phylogenetic relatedness of the strains are discussed. PMID- 9874229 TI - Neutral core oligosaccharides of bovine submaxillary mucin--use of lead tetraacetate in the cold for establishing branch positions. AB - Oligosaccharide alditols were released from bovine submaxillary mucin by alkaline borohydride treatment. The fractions containing smaller neutral oligosaccharides were separated by HPLC, to give, in the mono-, di- and trisaccharide-alditol sizes, 19 different structures, in addition to three fucosylated tetrasaccharide alditols. Molecules were identified by NMR spectroscopy, electrospray-mass spectrometry-mass-spectrometry, permethylation analyses, and highly selective Pb(OAc)4 oxidation at -30 degrees C, followed by borohydride reduction. Pb(OAc)4 oxidation was found to be generally applicable in identifying the branch position(s) of substitutions for all core structures. Among the isolated oligosaccharide alditols were structures not previously reported, including 12 structures not reported from bovine submaxillary mucin, and four structures (three core structures) not found in the Carbbank database. PMID- 9874232 TI - Amino-acid-sequence determination and biological activity of tessulin, a naturally occurring trypsin-chymotrypsin inhibitor isolated from the leech Theromyzon tessulatum. AB - We purified a new trypsin-chymotrypsin inhibitor, designated tessulin, from the rhynchobdellid leech Theromyzon tessulatum. This 9-kDa peptide was purified to apparent homogeneity by gel-permeation and anion-exchange chromatographies followed by reverse-phase HPLC. The structure of tessulin was determined by reduction, S-beta-pyridylethylation, trypsin digestion, automated Edman degradation and matrix-assisted laser desorption mass spectrometry (m/z 8985 Da). The 81-amino-acid peptide possesses 16 cysteines and exhibits a 16% sequence similarity with antistasin-type inhibitors. Tessulin inhibits trypsin (Ki 1 pM) and chymotrypsin (Ki 150 pM) and exhibits no activity with thrombin, factor Xa, cathepsin G and elastase. This is the first trypsin-chymotrypsin inhibitor isolated from leeches that does not inhibit elastase or cathepsin G, except for cytin and therin. Furthermore, tessulin, in conjunction with other serine protease inhibitors isolated from Theromyzon (therin, theromin), significantly diminishes the level of human granulocyte and monocyte activation induced by lipopolysaccharides (10 microg). The combined level of inhibition is higher than that of aprotinin, another serine-protease inhibitor used biomedically. Thus, tessulin may be clinically significant in reducing inflammatory events. PMID- 9874233 TI - Structure determination for octasaccharides derived from the carbohydrate-protein linkage region of chondroitin sulphate chains in the proteoglycan aggrecan from bovine articular cartilage. AB - Five octasaccharides derived from the protein carbohydrate linkage region of chondroitin sulphate (CS) have been isolated from the large aggregating proteoglycan (aggrecan) extracted from the bovine articular cartilage of 6-year old to 8-year-old animals. Following the purification of aggrecan the attached CS chains were digested with CS ABC endolyase and subsequently released from the protein core by beta-elimination. The individual oligosaccharides were purified by strong anion-exchange chromatography and their structures determined by very high-field one-dimensional and two-dimensional 1H-NMR spectroscopy. They were found to be octasaccharides, comprised of tetrasaccharide repeat-region extensions to the core tetrasaccharide linkage region structure. They have the following structures: deltaUA(beta1-3)GalNAc(beta1-4)GlcA(beta1-3)GalNAc(beta1 4)+ ++GlcA(beta1-3)Gal(beta1-3)Gal(beta1-4)Xyl-ol, deltaUA(beta1-3)GalNAc(beta1 4)GlcA(beta1-3)GalNAc6S(b eta1-4)GlcA(beta1-3)Gal(beta1-3)Gal(beta1-4)Xyl-ol, deltaUA(beta1-3)GalNAc6S(beta1-4)GlcA(beta1-3)GalNAc(b eta1-4)GlcA(beta1 3)Gal(beta1-3)Gal(beta1-4)Xyl-ol, deltaUA(beta1-3)GalNAc6S(beta1-4)GlcA(beta1 3)GalNA c6S(beta1-4)GlcA(beta1-3)Gal(beta1-3)Gal(beta1-4)Xyl-ol and deltaUA(beta1 3)GalNAc4S(beta1-4)GlcA(beta1-3)GalNA c6S(beta1-4)GlcA(beta1-3)Gal(beta1 3)Gal(beta1-4)Xyl-ol. They differ only in the nature of the sulphation of the GalNAc residues of the tetrasaccharide-repeat-region extension, which forms the first two disaccharides of the repeat region. No sulphation of any of the uronic acid residues has been identified and in one oligosaccharide neither of the GalNAc residues were sulphated. The majority of the linkage regions contained GalNAc residues which were fully 6-sulphated. However, in a significant amount, only one of the residues was 6-sulphated while the other was either unsulphated or 4-sulphated. There was no evidence either for sulphation of the linkage region galactose residues or for phosphorylation of the xylose residue, through which the chain is attached to the core protein. PMID- 9874234 TI - Glycan specificity of myelin-associated glycoprotein and sialoadhesin deduced from interactions with synthetic oligosaccharides. AB - Myelin-associated glycoprotein (MAG) and sialoadhesin (Sn) bind to sialylated glycans on cell surfaces and are thought to be involved in cell-cell interactions. In order to investigate how the interactions of these proteins are influenced by the glycan structure, we compared the inhibitory potencies of different synthetic monovalent oligosaccharides and polyvalent polyacrylamide derivatives. Using oligosaccharides with modifications in the sialic acid, galactose or N-acetylglucosamine moieties, we could demonstrate that both MAG and Sn bind with high preference to alpha2,3-linked sialic acid and interact at least with the three terminal monosaccharide units. For MAG, contacts with even more distant monosaccharides are likely, since pentasaccharides are bound better than trisaccharides. Also, an additional sialic acid at position six of the third terminal monosaccharide unit enhances binding to MAG, whereas it does not influence binding to Sn significantly. Modifications of the sialic acid glycerol side chain demonstrated that the hydroxy groups at positions 8 and 9 are required for binding to both proteins. Surprisingly, MAG binds 2-keto-3-deoxy-D-glycero-D galacto-nononic acid significantly better than N-acetylneuraminic acid, whereas Sn prefers the latter structure. These results indicate that the interactions of MAG and Sn are mainly with sialic acid and that additional contacts with the subterminal galactose and N-acetylglucosamine residues also contribute to the binding strength, although to a lesser degree. PMID- 9874235 TI - Biophysical characterisation of lysozyme binding to LPS Re and lipid A. AB - The binding of lysozyme to bacterial deep rough mutant lipopolysaccharide (LPS) Re and to its lipid moiety lipid A, the 'endotoxic principle' of LPS, was investigated using biophysical techniques. The beta<-->alpha gel to liquid crystalline phase transition, the nature of the functional groups of the endotoxins, the secondary structure of lysozyme, and competition with polymyxin B were studied by Fourier-transform infrared spectroscopy (FTIR); the supramolecular aggregate structure of the endotoxins was determined with synchrotron radiation X-ray diffraction and the binding stoichiometry with microcalorimetry. The results were compared with those found with zwitterionic and negatively charged phospholipids. It can clearly be shown that lysozyme binds electrostatically to charged groups of the endotoxin molecules with the consequence of acyl-chain rigidification and an initiation of a transition from inverted cubic to multilamellar structures. The binding stoichiometry of endotoxin and lysozyme is a 3:1 molar ratio for both LPS Re and lipid A, indicating a dominant binding of lysozyme to the lipid A-phosphates. This could be confirmed by the analysis of a phosphate vibration and by the use of a dephospho LPS. Parallel to lysozyme binding to endotoxin, a conformational change of the secondary structure in the protein from mainly alpha helix to more unordered structures takes place, while the residual beta-sheet substructure does not exhibit a clear concentration dependence. Binding is found to be specific for the endotoxins since, for the zwitterionic phosphatidylcholine, no binding is observed and, for the negatively charged phosphatidylglycerol, only very weak binding is found. The results are discussed in the context of the ability of lysozyme to reduce endotoxicity. PMID- 9874236 TI - Structural characterisation of a rhamnan and a fucorhamnan, both present in the lipopolysaccharide of Burkholderia vietnamiensis strain LMG 10926. AB - The polymeric fraction isolated after mild acid hydrolysis of the lipopolysaccharide (LPS) from Burkholderia vietnamiensis strain LMG 10926 contained L-rhamnose (Rha) and D-fucose (Fuc). From NMR studies supported by the results of methylation analysis and Smith degradation, it could be inferred that the material was probably a mixture of two glycans. One component was a linear rhamnan with a trisaccharide repeating unit (1); the other was a branched fucorhamnan with a tetrasaccharide repeating unit (2). The presence of two distinct polymeric fractions in LPS is a common feature for Burkholderia species. [structures: see text] PMID- 9874237 TI - Transport of arginine and ornithine into isolated mitochondria of Saccharomyces cerevisiae. AB - In this work we have characterised the transport of L-arginine and L-ornithine into mitochondria isolated from a wild-type Saccharomyces cerevisiae strain and an isogenic arg11 knock-out mutant. The Arg11 protein (Arg11p) is a mitochondrial carrier required for arginine biosynthesis [Crabeel, M., Soetens, O., De Rijcke, M., Pratiwi, R. & Pankiewicz, R. (1996) J. Biol. Chem. 271, 25011-25019]. Reconstitution experiments have confirmed that it is an L-ornithine carrier also transporting L-arginine and L-lysine by order of decreasing affinity, but not L histidine [Palmieri, L., De Marco, V., Iacobazzi, V., Palmieri, F., Runswick, M. & Walker, J. (1997) FEBS Lett. 410, 447-451]. Evidence is presented here that the mitochondrial inner membrane contains an L-arginine and L-ornithine transporting system distinct from Arg11p, in keeping with the arginine leaky phenotype of arg11 knock-out mutants. The newly characterised carrier, which we propose to name Bac1p (basic amino acid carrier), behaves as an antiporter catalysing the electroneutral exchange of the basic amino acids L-arginine, L-lysine, L ornithine and L-histidine and displays the highest affinity for L-arginine (Km of 30 microM). L-Arginine uptake has a pH optimum in the range of 7.5-9 and is inhibited by several sulphydryl reagents, by pyridoxal 5'-phosphate and by cations. PMID- 9874238 TI - Kinetic properties of a single nucleotide binding site on chloroplast coupling factor 1 (CF1). AB - The kinetics of nucleotide binding to spinach chloroplast coupling factor CF1 in a fully inhibited state were investigated by stopped-flow experiments using the fluorescent trinitrophenyl analogue (NO2)3Ph-ADP. The CF1 was in a state in which two of the three binding sites on the beta subunits were irreversibly blocked with ADP, Mg2+ and fluoroaluminate, while the three binding sites on the alpha subunits were occupied by nucleotides [Garin, J., Vincon, M., Gagnon, J. & Vignais, P. V. (1994) Biochemistry 33, 3772-3777)]. Thus, it was possible to characterise a single nucleotide-binding site without superimposed nucleotide exchange or binding to an additional site. (NO2)3Ph-ADP binding to the remaining site on the third beta subunit was characterised by a high dissociation rate of 15 s(-1), leading to a very low affinity (dissociation constant higher than 150 microM). Subsequent to isolation, CF1 preparations contained two endogenously bound nucleotides. Pre-loading with ATP yielded CF1 with five tightly bound nucleotides and one free nucleotide-binding site on a beta subunit. Pre-loading with ADP, however, resulted in a CF1 preparation containing four tightly bound nucleotides and two free nucleotide binding sites. One of the two free binding sites was located on a beta subunit, while the other was probably located on an alpha subunit. PMID- 9874239 TI - Structural heterogeneity in the O polysaccharide of Pseudomonas syringae pv. coriandricola GSPB 2028 (NCPPB 3780, W-43). AB - The O polysaccharide (OPS) of the lipopolysaccharide of Pseudomonas syringae pv. coriandricola GSPB 2028 (NCPPB 3780, W-43) was studied by Smith degradation and 1H-NMR and 13C-NMR spectroscopy, including two-dimensional COSY, TOCSY, NOESY, and H-detected 1H,13C heteronuclear multiple-quantum coherence (HMQC) experiments. The OPS was shown to consist of pentasaccharide O repeats of two types both containing four L-rhamnose and one 3-acetamido-3,6-dideoxy-D-galactose (D-Fuc3NAc) residue. Structure 1 of the major O repeat which had been established earlier [Das, S., Ramm, M., Kochanowski, H. & Basu, S. (1994) J. Bacteriol. 176, 6550-6557], was confirmed by our data, and a new structure 2 was elucidated for the minor O repeat and found to differ from the structure 1 only in the position of substitution of one of the rhamnose residues in the main chain. [structures: see text] A role of structural and immunochemical features of the LPS for defining the taxonomical position of the bacterium studied is discussed. PMID- 9874240 TI - Effects of histone and diolein on the structure of phosphatidylcholine/phosphatidylserine or phosphatidylcholine/phosphatidylglycerol bilayers. AB - The effects of the PKC substrate histone 1 and the PKC activator diolein (Ole2Gro) on the structure of phosphatidylcholine (PtdCho)/phosphatidylserine (PtdSer), or PtdCho/phosphatidylglycerol (PtdGro) bilayers were studied using 2H NMR. The results showed that in PtdCho/PtdSer bilayers, histone preferentially increased order parameters of the acyl chains of the PtdSer, but not the PtdCho lipid component. This effect was additive with the effect of Ole2Gro, which equally increased the ordering of the acyl chains of both PtdCho and PtdSer. The histone-induced change in the conformation of the PtdCho headgroups in PtdCho/PtdSer bilayers indicated that positively charged residues of the bound histone are located above the lipid-water interface and their location was altered by the presence of Ole2Gro. A different picture was observed in the case of PtdCho/PtdGro bilayers; although the effect of Ole2Gro on both the PtdCho or the PtdGro components was similar to the case of the PtdCho/PtdSer bilayers, histone did not significantly affect the order parameters of PtdCho or PtdGro in either the absence or presence of Ole2Gro. The results indicate that histone 1 induces clustering of PtdSer in PtdCho bilayers which may contribute to PKC activation. Moreover, the observed differences in the interactions of histone with PtdCho/PtdSer compared with PtdCho/PtdGro bilayers may explain the higher efficiency of PtdSer in activating PKC. PMID- 9874241 TI - Cyclophilin-D binds strongly to complexes of the voltage-dependent anion channel and the adenine nucleotide translocase to form the permeability transition pore. AB - A cyclophilin-D affinity matrix was employed to isolate components of the mitochondrial permeability transition pore. A cDNA encoding cyclophilin-D was cloned from a rat liver library and ligated into pGEX to allow expression of a glutathione S-transferase/cyclophilin-D fusion protein in Escherichia coli XL1 cells. The cyclophilin-D in the fusion was functionally normal as judged by its peptidylprolyl cis-trans-isomerase activity and its inhibition by cyclosporin A. The fusion protein was bound to glutathione-agarose to form the cyclophilin-D affinity matrix. The matrix selectively bound 32-kDa proteins of mitochondrial membrane extracts, but no H2O-soluble proteins were bound. The 32-kDa band on SDS/PAGE resolved into a doublet and reacted with antibodies against the voltage dependent anion channel (porin) and the adenine nucleotide translocase. These two proteins were also selectively retained by the affinity matrix in the presence of cyclosporin A. The thus-purified voltage-dependent anion channel, adenine nucleotide translocase and the fusion protein were incorporated into phosphatidylcholine liposomes containing fluorescein sulphonate. The proteoliposomes were permeabilized by Ca2+ plus phosphate, and this was blocked completely by cyclosporin A. These properties are identical to those of the permeability transition pore in mitochondria. It is concluded that the basic permeability transition pore structure comprises the voltage-dependent anion channel (outer membrane), adenine nucleotide translocase (inner membrane) and cyclophilin-D, and forms at contact sites between the two membranes. PMID- 9874242 TI - Genes of succinyl-CoA ligase from Saccharomyces cerevisiae. AB - Succinyl-CoA ligase (succinyl-CoA synthetase) catalyzes the nucleotide-dependent conversion of succinyl-CoA to succinate. This enzyme functions in the tricarboxylic acid (TCA) cycle and is also involved in ketone-body breakdown in animals. The enzyme is composed of alpha and beta subunits that are required for catalytic activity. Two genes, LSC1 (YOR142W) and LSC2 (YGR244C), with high similarity to succinyl-CoA ligase subunits from other species were isolated from Saccharomyces cerevisiae. The expression of these genes was repressed by growth on glucose and was induced threefold to sixfold during growth on nonfermentable carbon sources. The LSC genes were deleted singly and in combination. Unlike other yeast strains with defects in TCA cycle genes, strains lacking either or both LSC genes were able to grow with acetate as a carbon source. However, growth on glycerol or pyruvate was impaired. An antiserum against both subunits of the Escherichia coli enzyme was capable of recognizing the yeast succinyl-CoA ligase alpha subunit, and this band was absent in delta lsc1 deletion strains. Succinyl CoA ligase activity was absent in mitochondria isolated from strains deleted for one or both LSC genes, but activity was restored by the presence of the appropriate LSC gene on a plasmid. The yeast succinyl-CoA ligase was shown to utilize ATP but not GTP for succinyl-CoA synthesis. PMID- 9874243 TI - Involvement of NF-Y in transcriptional regulation of the phospholamban gene. AB - To understand the transcriptional regulation of the phospholamban gene, we analyzed a 5'-upstream region of the gene. Using a series of deletion constructs, we demonstrated that the region from -96 bp to -78 bp, containing the CCAAT sequence, is essential for transcription of this gene. This region specifically bound to nuclear proteins extracted from rat hearts, and gel-shift assays using competitive oligonucleotides, antibodies and recombinant proteins showed that this region binds to the NF-YA and NF-YB, members of the CCAAT-binding nuclear protein family. This region-dependent transcription in cardiac myocytes transfected with antisense cDNAs encoding NF-YA and NF-YB was decreased to approximately 50% of that seen in cells transfected with the same sense cDNAs. We, therefore, conclude that the region from -96 bp to -78 bp plays a critical role in expression of the phospholamban gene, which is regulated by binding of the nuclear protein NF-Y. PMID- 9874244 TI - Cloning and sequence analysis of cDNAs encoding human placental tissue protein 17 (PP17) variants. AB - Using monospecific anti-PP17 serum with chemiluminescence Western-blot analysis, we detected different molecular-mass variants of human soluble placental tissue protein 17 (PP17) in different normal adult and fetal human tissues besides term placenta. 13 cDNAs with three different insert lengths encoding PP17 variants were isolated by screening a human placental cDNA library. Sequence analysis of the shortest clones showed that the inserts contain the same open reading frame encoding PP17a variant (28,129 kDa) consisting of 251 residues, which is identical to the previously isolated and characterised PP17 antigen described in 1983. The ubiquitous PP17b variant is encoded by longer clones and contains 434 residues with a predicted molecular mass of 47,208 kDa. Compared to normal conditions, these newly discovered PP17 variants are overexpressed in cervix carcinoma tissue, as are their three different-size messenger RNAs in HeLa cell line. Increased amounts of PP17b are secreted into the circulation in cervix carcinoma patients. We also observed a typical elevation in serum levels of PP17 variants during healthy pregnancy. An alignment search of the protein databank showed that PP17a and PP17b are homologous to adipose tissue differentiation and lipid-droplet-associated proteins: human adipophilin, mouse adipose differentiation-related protein and rat perilipin A and B. PMID- 9874245 TI - Structural determinants in 5S RNA and TFIIIA for 7S RNP formation. AB - C2H2-type zinc-finger modules define a unique structural motif, which is capable of forming specific complexes with both DNA and RNA. While the principles governing DNA binding have been defined in great detail, the mode of RNA recognition remains only poorly understood. In the absence of information from three-dimensional structural analysis of a zinc-finger/RNA complex, we have performed a number of biochemical studies to gain further insight into the molecular details of the interaction of 5S ribosomal RNA with the zinc-finger protein TFIIIA. Previous work had indicated that zinc finger 6 of TFIIIA contacts 5S RNA in close proximity or directly in the loop-A region (nucleotides 10-13). Permutation analysis of this sequence reveals that three of the four nucleotides are of vital importance for RNA recognition. Exchange of unusual and therefore characteristic aromatic residues in finger 6 against aliphatic or other aromatic amino acids reveals that the aromatic character of tryptophan 177 is essential for RNA recognition. Association with helix V in 5S RNA appears to involve specific contacts with the phosphate backbone, as evidenced by ethylation interference assays. Introduction of multiple internal and 3'-terminal as well as 5'-terminal deletions accompanied by stabilizing sequence substitutions defines a minimal RNA fragment that is sufficient for TFIIIA binding. This RNA molecule includes a truncated/mutated helix I, helix II and helix V, as well as structurally intact loops A and E. Permutation analysis of the loop-E region emphasizes its importance for TFIIIA recognition. PMID- 9874246 TI - Molecular cloning and bacterial expression of a general odorant-binding protein from the cabbage armyworm Mamestra brassicae. AB - A cDNA clone encoding a general odorant-binding protein (GOBP2) was isolated from antennal RNA of Mamestra brassicae by reverse transcription-PCR (RT-PCR) and RACE PCR. The cDNA encoding the GOBP2 was further used for bacterial expression. Most of the recombinant GOBP2 (>90%) was found to be insoluble. Purification under denaturing conditions consisted of solubilisation of inclusion bodies, affinity chromatography, refolding and gel filtration. The refolded rGOBP2 was cross reactive with a serum raised against the GOBP2 of the Lepidoptera Antheraea polyphemus. The purified refolded rGOBP2 was further characterised by native PAGE, IEF, N-terminal sequencing, and two-dimensional NMR. A functional characterisation of the rGOBP2 was carried out by testing its ability to bind pheromone compounds. The yields of production and purification fulfil the requirements of structural studies. PMID- 9874247 TI - Structural determination of sulfated tetrasaccharides and hexasaccharides containing a rare disaccharide sequence, -3GalNAc(4,6-disulfate)beta1-4IdoAalpha1 , isolated from porcine intestinal dermatan sulfate. AB - In the course of structural studies on sulfated oligosaccharides isolated from porcine intestinal heparin after extensive digestion with Flavobacterium heparinase, we isolated several heparitinase-resistant unsaturated oligosaccharides. Amino sugar analysis of these oligosaccharides indicated that they contained galactosamine residues but no glucosamine residues. They were sensitive to chondroitinase ABC but resistant to chondroitinase AC-II, and therefore derived from dermatan sulfate, which was presumably contained as a minor component in the starting heparin preparation. The structures of these oligosaccharides were characterized by enzymatic digestions in conjunction with HPLC analysis of the digests and by one-dimensional and two-dimensional 500-MHz 1H-NMR spectroscopy. Structures of two tetrasaccharides and two hexasaccharides were determined as deltaHexAalpha1-3GalNAc(4S)beta1-4IdoAalpha1-3GalNAc(4S), deltaHexAalpha1-3GalNAc(4S,6S)]beta1-4IdoAalpha1-3GalNAc(4S) , deltaHexAalpha1 3GalNAc(4S)beta1-4IdoAalpha1-3GalNAc(4S)beta 1-4IdoAalpha1-3GalNAc(4S), and deltaHexAalpha1-3GalNAc(4S)beta1-4IdoAalpha1-3GalNAc(4S,6S)b eta1-4IdoAalpha1 3GalNAc(4S), where deltaHexA, IdoA, GalNAc, 4S and 6S represent 4-deoxy-alpha-L threo-hex-4-enepyranosyluronic acid, L-iduronic acid, N-acetyl-D-galactosamine, 4 O-sulfate and 6-O-sulfate, respectively. The latter three compounds have never been reported as discrete structures. Since the four isolated oligosaccharides contained an unsaturated uronic acid residue at the nonreducing terminus, they appear to have been generated by eliminative cleavage by the action of Flavobacterium chondroitinase that was probably present as a minor contaminant in the Flavobacterium heparinase preparation used. Two out of the four oligosaccharides shared the rare disulfated disaccharide sequence, 3GalNAc(4S,6S)beta1-4IdoAalpha1-. These oligosaccharides will be useful as authentic reference compounds for microanalyzing biologically active domains of dermatan sulfate. PMID- 9874248 TI - Selective coupling of STAT factors to the mouse prolactin receptor. AB - Prolactin has been reported to induce distinct sets of signal transducers and activators of transcription (STAT) in a cell-type-specific fashion. In the mammary epithelium, although STAT1, STAT3, STAT5A, STAT5B and STAT6 are present in a latent form, only STAT5A and STAT5B are activated. This selective activation of STAT5 by prolactin was also observed in COS-7 cells cotransfected with the long form of the mouse prolactin receptor (PRL-R) and expression vectors for STAT1, STAT3, STAT5 and STAT6. Mutated PRL-Rs and chimeric erythropoietin/gp130 (EPO/gp130) receptors with a tyrosine-containing motif attached at the carboxy terminus were employed to determine the sites in the PRL-R required for the specific activation of STAT5. The experiments revealed the importance of two motifs containing Y477 and Y578 in the PRL-R. When linked to the EPO/gp130 receptor, these sequences were sufficient to specifically induce DNA binding of STAT5 and to activate transcription from the beta-casein gene promoter. By contrast, only weakly they induced DNA binding of STAT6 and STAT3 and did not induce STAT1. A synthetic nonapeptide with phosphorylated Y477 was able to disrupt STAT5 DNA binding in vitro. Our results define structural domains within the carboxy terminus of the PRL-R which recruit STAT5 for signalling and which are capable of distinguishing STAT5 from other STAT proteins. The activity of STAT5 forms with deletions of the carboxy terminus was induced more strongly than that of their full-length counterparts 2 min after activation of the PRL-R. This effect was not dependent on the presence of Y477 and Y578 in the PRL-R, indicating that facilitated activation of short STAT5 isoforms relies on mechanisms other than increased coupling to specific regions of the PRL-R. PMID- 9874250 TI - Functional analysis of the human cytochrome P4501A1 (CYP1A1) gene enhancer. AB - The environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or dioxin) induces gene transcription, a process that requires binding of the activated aryl hydrocarbon receptor (AhR) to dioxin-responsive elements (DREs) within the enhancer region of responsive genes. Most of what is known about the molecular mechanism of AhR-dependent gene activation results from studies on the murine prototype TCDD-responsive gene cytochrome P4501A1 (CYP1A1). Much less is known, however, about the regulation of human TCDD-responsive genes. We have therefore conducted a detailed analysis of the enhancer region of the human CYP1A1 gene. From the ten DRE core motifs investigated within a stretch of 1400 bp in two human tumor cell lines using a ligation-mediated PCR technique, five motifs displayed a TCDD-inducible in vivo footprint. Four of these sites were functional enhancer sequences as demonstrated by a transient expression assay. Based on these data, a distinct functional consensus sequence for DRE motifs within the human CYP1A1 gene is suggested. After introduction of the four functional sites into various mouse hepatoma cell lines, only three exhibited a functional response, suggesting some species differences in CYP1A1 gene regulation. In addition to the footprints at DRE sites, we also detected protein DNA interactions at three G-rich domains located within the enhancer region of the human CYP1A1 gene. Our data show that, besides some similarities in the regulation of the human and mouse CYP1A1 genes, there also exist some distinct differences, including number, location, and functional consensus sequences of DRE motifs, as well as quantity and location of footprinted G-rich domains. PMID- 9874249 TI - Purification and cDNA cloning of cytokinin-specific binding protein from mung bean (Vigna radiata). AB - Synthetic urea derivatives such as N-phenyl-N'-(4-pyridyl)urea (4PU) and N-(2 chloro-4-pyridyl)-N'-phenylurea (4PU30) have strong cytokinin activities. Using tritiated 4PU30 as a probe, we previously established the presence of a cytokinin specific binding protein (CSBP) of high affinity (Ka for 4PU30 = 4x10(10) M(-1)) in the soluble fraction of etiolated mung bean seedlings [Nagata, R., Kawachi, E., Hashimoto, Y. & Shudo, K. (1993) Biochem. Biophys. Res. Commun. 191, 543 549]. In this report, we purified CSBP by the use of 4PU-Sepharose 4B, an affinity gel liganded with 4PU. We determined partial amino acid sequences of CSBP and isolated its cDNA by reverse-transcription (RT) PCR. The cDNA encoded a protein with a calculated molecular mass of 17 kDa. A data base homology search revealed that CSBP is a novel member of a major pollen allergen/pathogenesis related protein family. Recombinant CSBP was expressed in Escherichia coli and was confirmed to bind specifically to cytokinins. PMID- 9874251 TI - Glutamate dehydrogenase, the marker protein of Plasmodium falciparum--cloning, expression and characterization of the malarial enzyme. AB - The gene of an NADP+-specific glutamate dehydrogenase was cloned from Plasmodium falciparum, the causative agent of tropical malaria. Southern-blot analysis indicates a single-copy gene. The gene encodes a protein with 470 residues which has 50% of all residues identical with those of the glutamate dehydrogenases from other low eukaryotes and eubacteria. In contrast, the sequence identity with the human enzyme is marginal, which underlines the long evolutionary distance between parasite and host. The gene was overexpressed in Escherichia coli. The kinetic properties of the recombinant enzyme are in good agreement with those of the authentic enzyme. The parasite enzyme is inhibited by D-glutamate and glutarate, but not by chloroquine. Like other coenzyme-specific glutamate dehydrogenases, but in contrast to the dual-specific mammalian enzymes, the P. falciparum enzyme is not affected by GTP and ADP. The physical and chemical properties of the protein are in accordance with the cytosol being the major localization. The gene does not encode a cleavable mitochondrial presequence and the Mr of the recombinant protein and the protein isolated from the parasite are indistinguishable on SDS/PAGE. Western-blot analysis of stage-specific parasites shows that glutamate dehydrogenase is present in all intraerythrocytic stages. The signal increased continuously from rings, early trophozoites to late trophozoites and decreased slightly in the segmenter stage. Glutamate dehydrogenase, suggested to be the major source of NADPH in the parasite, is an attractive target molecule for the rational development of new antimalarial drugs. PMID- 9874252 TI - Heme-regulated eIF-2alpha kinase purifies as a hemoprotein. AB - The regulation of protein synthesis by the availability of heme in reticulocytes is well established. However, the mechanism by which heme regulates translational initiation is not clear. In this study, we have examined the heme regulation directly on the homogeneous heme-regulated eIF-2alpha kinase (HRI), which is activated during heme deficiency. We found that HRI purified as a hemoprotein with the characteristic Soret band of hemoprotein at 424 nm. This HRI was an active autokinase and eIF-2alpha kinase, and its kinase activities were inhibited by submicromolar concentrations of hemin with an apparent Ki of 0.5 microM. Homogeneous HRI was a homodimer, and its activities could not be inhibited by incubation with purified inactive K199R HRI in vitro. Our results suggest that there are two distinct types of heme-binding sites in the HRI homodimer. The binding of heme to the first site is stable, while the binding of heme to the second site is responsible for the rapid downregulation of HRI activity by heme. These results indicate that HRI binds heme and serves as a sensor of the availability of heme to coordinate the balanced synthesis of globins and heme in erythroid cells. PMID- 9874253 TI - Crystals of a 1:1 complex between human interleukin-4 and the extracellular domain of its receptor alpha chain. AB - The specific high-affinity binding of interleukin-4 (IL-4) to its receptor alpha chain is the crucial primary event during IL-4 signalling. Single crystals, suitable for high resolution diffraction studies, have been obtained from a complex between IL-4 and the ectodomain of the receptor alpha chain, also called IL-4-binding protein (IL-4BP). The orthorhombic crystals are in spacegroup P2(1)2(1)2(1) with cell constants a = 5.038 nm, b = 6.841 nm, c = 10.95 nm and diffract to a resolution of at least 0.25 nm when exposed to synchrotron radiation. The volume of the unit cell suggests the presence of a 1:1 IL-4/IL-4BP complex and HPLC analysis of the crystals confirmed that IL-4 and IL-4BP were present in equimolar amounts. An IL-4 variant comprising a total of four methionine residues was generated, labelled with selenomethionine and crystallised in complex with IL-4BP. The crystals are isomorphous to that of the complex with normal IL-4 and therefore can be used to solve the crystallographic phase problem by the method of multiple anomalous diffraction (MAD). The crystal structure of the IL-4/IL-4BP complex will help to understand how IL-4 and other helical cytokines bind and activate their cognate receptor. PMID- 9874254 TI - Recombinant homo- and hetero-oligomers of an ultrastable chaperonin from the archaeon Pyrodictium occultum show chaperone activity in vitro. AB - The archaeon Pyrodictium occultum is one of the most thermophilic organisms presently known. Previous experiments provided support for the significant contribution of a high-molecular-mass protein complex to the extreme thermotolerance of P. occultum. This protein complex, the 'thermosome', is composed of two subunits, alpha and beta, which form a hexadecameric double ring complex. In order to obtain the thermosome in amounts sufficient for structural and functional investigations, we produced the two subunits jointly and separately in Escherichia coli BL21(DE3). In all three cases, we isolated soluble, high-molecular-mass double-ring complexes from E. coli BL21(DE3). On electron micrographs, the recombinant complexes were indistinguishable from each other and from the natural thermosome. To characterize the quaternary structure of the recombinant particles, we used native gel electrophoresis, analytical gel filtration, and analytical ultracentrifugation. Spectral analysis, using absorption, fluorescence emission and far-UV circular dichroism spectroscopy were applied to compare the three recombinant protein complexes with the natural thermosome from P. occultum. All three recombinant complex species exhibit ATPase activity. Furthermore, we could demonstrate that the recombinant complexes slow down the aggregation of citrate synthase, alcohol dehydrogenase, and insulin. Thus, we conclude that the recombinant protein complexes exhibit a chaperone-like activity, interacting with non-native proteins; they do so at temperatures far below the lower physiological limit of growth. PMID- 9874255 TI - Enhancement of phosphoinositide 3-kinase (PI 3-kinase) activity by membrane curvature and inositol-phospholipid-binding peptides. AB - The phosphorylation of phosphatidylinositol (PtdIns) on the 3' position of the inositol ring by phosphoinositide 3-kinase (PI 3-kinase) is shown to depend strongly on the curvature of liposomes containing a mixture of phosphatidylcholine (PtdCho) and PtdIns. Vesicles with an average diameter of 50 nm are phosphorylated 100 times faster than chemically identical vesicles with an average diameter greater than 300 nm. The low reactivity of large vesicles is not due to the difference in vesicle number for large and small vesicles at constant total lipid, nor to occlusion of lipid surfaces in multilammelar structures, and can be reversed by addition of low (< 1:100) molar ratios of either the PtdIns transfer protein sec14p or a ten-residue peptide derived from the inositol phospholipid-binding site of gelsolin. Similar measurements using PI 4-kinase showed a weak dependence on vesicle size. The strong dependence of PI 3-kinase function on membrane curvature suggests possible localization of PI 3-kinase activity at sites where clustering of receptors, for example, may locally deform the membrane, and suggests that once PI 3-kinase is localized and activated at surface sites, the reaction may become self-accelerating. PMID- 9874256 TI - The roles of the N-linked glycans and extension regions of soybean beta conglycinin in folding, assembly and structural features. AB - Beta-conglycinin, one of the dominant storage proteins of soybean, is a trimer composed of three subunits, alpha, alpha' and beta. All subunits are N glycosylated and alpha and alpha' contain extension regions in addition to the core regions common to all subunits. Non-glycosylated individual subunits and deletion mutants (alpha(c) and alpha'(c)) lacking the extension regions of alpha and alpha' were expressed in Escherichia coli. All recombinant proteins were purified to near homogeneity and appeared to have the correct conformation, as judged by CD, density-gradient centrifugation and gel-filtration profiles, indicating that the N-linked glycans and extension regions are not essential for the folding and the assembly into trimers of beta-conglycinin. Density-gradient centrifugation, gel-filtration and differential scanning calorimetry profiles of the recombinant proteins and the native beta-conglycinin indicated that the N linked glycans and extension regions contribute to the dimension of beta conglycinin but not to the density and the thermal stability. Comparing the solubilities of the individual subunits with those of deletion mutants, only the alpha and alpha' subunits were soluble at lower ionic strength (mu < 0.25) at around the pH value of the endoplasmic reticulum. This suggests that the extension regions play an important role in the prevention of aggregation in the endoplasmic reticulum in analogy with the N-linked glycans. PMID- 9874257 TI - Purification, cDNA cloning and expression of human NG,NG-dimethylarginine dimethylaminohydrolase. AB - cDNA encoding N(G),N(G)-dimethylarginine dimethylaminohydrolase from rat kidney had been cloned [Kimoto, M., Sasakawa, T., Tsuji, H., Miyatake, S., Oka, T., Nio, N. & Ogawa, T. (1997) Biochim. Biophys. Acta 1337, 6-10]. The enzyme hydrolyzes N(G),N(G)-dimethyl-L-arginine and N(G)-monomethyl-L-arginine, which are known as endogenous inhibitors for the nitric oxide-generating system. In the present study, human N(G),N(G)-dimethylarginine dimethylaminohydrolase has been purified to homogeneity from liver and characterized. The cDNA clone encoding human N(G),N(G)-dimethylarginine dimethylaminohydrolase was isolated from a human kidney lambda gt10 library using a probe prepared from a plasmid containing the entire coding region of rat N(G),N(G)-dimethylarginine dimethylaminohydrolase. Its open reading frame encoded a protein of 285 amino acids with a molecular mass of 31,121 Da. The deduced amino acid sequence exhibits 93% identity with that of rat. The cDNA was expressed as a fusion protein in Escherichia coli and the recombinant protein exhibited enzyme activity which is the same as that of natural enzyme. PMID- 9874258 TI - Structure/function relationship of CYP11B1 associated with Dahl's salt-resistant rats--expression of rat CYP11B1 and CYP11B2 in Escherichia coli. AB - Dahl's salt-resistant normotensive rats (DR rats) have been previously reported to express cytochrome P-450 (CYP11B1) containing five missense mutations [Matsukawa, N., Nonaka, Y., Higaki, J., Nagano, M., Mikami H., Ogihara, T. & Okamoto, M. (1993) J. Biol. Chem. 268, 9117-9121]. To investigate structure function relationships of CYP11B, wild-type rat CYP11B1 and CYP11B2 and DR CYP11B1 (mutant CYP11B1 in Dahl's salt-resistant rats) have been successfully expressed in Escherichia coli. Steroid 11beta-hydroxylase (11beta-OHase) activity observed with DR-CYP11B1 was similar to that of wild-type CYP11B1, while 18 hydroxylase (18-OHase) activity of DR-CYP11B1 was lower than that of wild-type CYP11B1. Mutant CYP11B1s containing a single or a double amino acid substitution associated with DR-CYP11B1 have been also expressed in E. coli to investigate effects of the substitutions on enzymatic activity. Each of the single mutant enzymes showed lower 18-OHase activity than wild-type CYP11B1, but not as low as DR-CYP11B1. A double mutant CYP11B1 with V381L and I384L showed 18-OHase activity at a similar low level to that of DR-CYP11B1. The 19-hydroxylation (19-OHase) activity of DR-CYP11B1 was about one-third of that of the wild-type enzyme and this low activity appeared due to the V443M mutation. These results suggest that three of five amino acid substitutions present in DR-CYP11B1 account for the decreased 18-OHase and 19-OHase activities. A decrease in these enzyme activities may be responsible for the normotension of the DR rats when fed a high-salt diet. PMID- 9874259 TI - Both Ets-1 and GATA-1 are essential for positive regulation of platelet factor 4 gene expression. AB - In the rat platelet factor 4 (PF4) promoter, Ets motifs and GATA motifs are located at positions -880, -75 and -135, -30, respectively, and their motifs are found in the promoter region of most megakaryocyte protein genes. In order to investigate how the Ets and GATA motifs affect PF4 promoter activity, we constructed Ets and/or GATA motif mutant genes. A single disruption of either 75Ets, -135GATA, or -30GATA significantly reduced PF4 promoter activity, and double disruptions involving these motifs completely abolished it. Furthermore, gel-retardation assays revealed that Ets-1 and GATA-1 proteins from HEL and MEG 01 cells bound to the Ets motifs and GATA motifs, respectively. Co-transfection experiments showed that the overexpression of Ets-1 and/or GATA-1 enhanced the expression of the PF4 promoter reporter gene. These effects of Ets-1 and GATA-1 on PF4 promoter activity are additive. When HEL cells were treated with dimethylsulfoxide in order to induce differentiation into megakaryocytes, the mRNA level of ets-1 increased 10-fold, which might be directly correlated with the significant increase in PF4 mRNA level induced by dimethylsulfoxide. All these results strongly suggest that both Ets-1 and GATA-1 play key roles in the positive regulation of PF4 gene expression. PMID- 9874260 TI - Protein LA, a novel hybrid protein with unique single-chain Fv antibody- and Fab binding properties. AB - Existing Ig-binding proteins all suffer from limitations in their binding spectrum. In the pursuit of the ultimate, non-restricted, Ig-binding protein, we have constructed the hybrid protein LA, by fusing four of the Ig kappa light chain-binding domains of peptostreptococcal protein L with four of the IgGFc- and Fab-binding regions of staphylococcal protein A. Ligand-blot experiments demonstrated that the L and the A components were both functional in the hybrid, as the protein was shown to bind purified kappa light chains and IgGFc. Protein LA bound human Ig of different classes and IgG from a wide range of mammalian species. IgG, IgM and IgA were purified from human serum and saliva by affinity chromatography on protein LA agarose. Similarly, single-chain Fv (scFv) antibodies carrying the kappa light-chain variable domain or expressing the V(H)III (variable domain of the heavy chain of Ig) determinant, were efficiently purified on immobilized protein LA. As judged by surface plasmon resonance (SPR), protein LA showed enhanced affinity for all tested ligands, including several scFv antibodies, compared with proteins L and A alone. SPR analysis also demonstrated that binding of a ligand to one of the components in protein LA did not affect the ability of the hybrid protein to interact simultaneously with a ligand for the other component. The antigen-binding capacity of a kappa expressing scFv antibody was unaffected by the interaction with protein LA, whereas the binding of a V(H)III-expressing scFv antibody to its antigen was, unexpectedly, blocked by protein A and protein LA. Together, these data demonstrate that protein LA represents a highly versatile Ig-binding molecule. PMID- 9874261 TI - Multiple specificities of the staphylococcal and streptococcal fibronectin binding microbial surface components recognizing adhesive matrix molecules. AB - Many pathogenic gram-positive bacteria express fibronectin (Fn)-binding microbial surface components recognizing adhesive matrix molecules (MSCRAMMs), most of which have a similar structural organization with a primary ligand-binding domain consisting of 3-6 repeats of 40-50 amino-acid-residue motifs. The MSCRAMMs appear to preferentially bind to the N-terminal region of Fn, which is composed of five type-I modules. Here we report that the Fn-binding MSCRAMM FnbpA of Staphylococcus aureus contains a second ligand-binding domain located outside the repeat units. In addition, several sites in the Fn N-terminus presented as recombinant type-I module pairs bind to the repeat domain of the MSCRAMM. All of the MSCRAMMs analyzed, which include FnbpA of Staphylococcus aureus, Sfb of Streptococcus pyogenes, and FnbA and FnbB of Streptococcus dysgalactiae, were shown to bind to multiple sites in the N-terminal domain of Fn. By dissecting the repeat domain of FnbpA using synthetic peptides and recombinant fragments, we show that discrete, different motifs are responsible for the binding to individual sites in Fn, rather than a common motif being able to bind to several pairs of type-I Fn modules. The C-terminal half of many of the MSCRAMM repeat units contain a common motif, which is shown here to bind to the type-I module pair 4 and 5. In addition, some of the repeat units of FnbpA contain N-terminal motifs which bound to the type-I module pairs 1-2 and 2-3, respectively. These latter binding motifs appear to be partly overlapping and dependent on flanking sequences. Fluorescence polarization experiments using fluorescein-labeled MSCRAMM peptides and recombinant type-I Fn module pairs revealed dissociation constants of 1-13 microM. It was also shown that the fluorescein-labeled peptides differed in their primary binding sites on Fn. PMID- 9874262 TI - The sulphoxidation of thioanisole catalysed by lactoperoxidase and Coprinus cinereus peroxidase: evidence for an oxygen-rebound mechanism. AB - Using both stopped-flow and conventional spectroscopy, the oxygenation of methyl phenyl sulphide by both lactoperoxidase (LPO) and Coprinus cinereus peroxidase (CiP) was monitored. Controlled continuous addition of H2O2 during turnover and monitoring the presence of native enzymes, compounds I, II and III, led to formation of the sulphoxide in high yield and enantioselectivity. Under those conditions, LPO catalysed the formation of (R) methyl phenyl sulphoxide with a yield of 85% and an enantiomeric excess (e.e.) of 80%. CiP catalysed the formation of (S) methyl phenyl sulphoxide with a yield of 84% and an e.e. of 73%. The enantioselective performance was markedly influenced by the purity of the enzymes used. Presence of compound III during turnover led to rapid inactivation of the peroxidases and, therefore, to both a lower yield of the sulphoxides and a lower enantioselectivity. Stopped-flow kinetic data show that, for both LPO and CiP, the transition of compound I to compound II depends on the concentration of the methyl phenyl sulphide, suggesting an oxygen-rebound mechanism. In line with this mechanism, a methyl phenyl sulphide radical cation was detected by EPR during turnover for LPO. PMID- 9874263 TI - Tissue-specific, cell cycle-regulated chimeric transcription factors for the targeting of gene expression to tumor cells. AB - A major challenge in the gene therapy of proliferative diseases is the specific targeting of gene expression. Here we describe a new approach based on the development of dual-specificity promoters that are both cell type specific and cell cycle regulated. The gene of interest is driven by an artificial heterodimeric transcription factor, whose DNA-binding subunit is expressed from a tissue-specific promoter, whereas the trans-activating subunit is transcribed from a cell cycle-regulated promoter. As a result gene expression occurs preferentially in the proliferating cells of a specific type of tissue. The selectivity of this strategy is demonstrated for the expression of a transgene in proliferating melanoma cells, using a combination of cyclin A and tyrosinase promoter elements. We also show that the level of expression that can be achieved by this system is sufficient to induce a clear biological effect in a TNF-alpha cytotoxicity assay. PMID- 9874265 TI - Targeted in vivo delivery of therapeutic gene into experimental squamous cell carcinomas using anti-epidermal growth factor receptor antibody: immunogene approach. AB - The "Fab immunogene" is a novel gene transfer vehicle in which the Fab fragment of anti-human epidermal growth factor (EGF) receptor antibody B4G7 is conjugated with poly-L-lysine to form an affinity complex with DNA. It was developed to target delivery of therapeutic genes into EGF receptor-hyperproducing tumor cells. Various characteristic features of the immunogene have been documented (Chen et al., 1998). Here we add further evidence to prove that in vitro transfer of beta-galactosidase/Fab immunogene is exclusively to EGF receptor-positive cells and that the herpes simplex virus thymidine kinase (TK)/Fab immunogene induces substantial suicide effects on A431 tumor cells when treated together with ganciclovir. The in vivo specificity of the immunogene transfer was examined using A431 tumor-bearing nude mice. When these nude mice were injected intraperitoneally with the chloramphenicol acetyltransferase (CAT)/Fab immunogene, CAT DNA was detected in the tumors as well as in liver and kidney but not brain, whereas CAT mRNA and enzyme activity were detected only in the tumors. Local and intraperitoneal injection of the TK/Fab immunogene and subsequent administration of ganciclovir effectively suppressed the growth of A431 tumors transplanted on the backs of nude mice. These observations suggest a possible application of the Fab immunogene system in cancer gene therapy. PMID- 9874264 TI - Enhanced in vivo airway gene transfer via transient modification of host barrier properties with a surface-active agent. AB - Effective adenoviral gene therapy requires efficient viral vector entry into epithelial cells. Injured airway epithelia display enhanced gene transfer, reflecting in part increased vector access to protected cell populations and/or protected basolateral membranes. We tested whether adenoviral gene transfer is enhanced by modification of the epithelial barrier in mouse nasal airways with a nonionic detergent (polidocanol, PDOC). In C57BL/6 mice, 1.6 x 10(9) PFU of Ad5CMV LacZ (AdLacZ) instilled into the right nostril produced negligible gene transfer to the nasal epithelium 2 days after dosing, but significant, dose dependent increases in gene transfer were achieved by pretreatment with PDOC. Permeation of the electron-dense tracer lanthanum into the intercellular junctions of PDOC (0.1%)-treated murine nasal epithelium, but not into intercellular junctions of vehicle controls, is consistent with PDOC-mediated increases in tight junctional permeability. In CF(-/-) mice, significant gene expression was not detectable after exposure to Ad5CBCFTR alone (1.4 x 10(9) PFU in 20 microl; AdCFTR), but PDOC pretreatment prior to AdCFTR instillation produced functional expression of CFTR (measured as deltaPD) 5 days after instillation. Because the development and testing of lung gene therapy will principally occur in children and adults with airway disease, AdLacZ gene transfer with and without PDOC pretreatment was examined in infected nasal airways. Gene expression was significantly reduced in infected as compared with uninfected airways. We conclude that the use of adjuvant surface-active and/or membrane-perturbing agents, synthetic or naturally derived, may provide a novel approach to enhancing the efficiency of adenoviral gene transfer. PMID- 9874266 TI - Construction, propagation, and titer estimation of recombinant adenoviruses carrying proapoptotic genes. AB - Generation of a recombinant adenovirus (Adv) that induces the constitutive expression of an apoptotic gene has been extremely difficult owing to severe apoptotic damage to the host cell. In this study, 293 cells were transduced with the caspase-inhibiting CrmA gene (293-CrmA cells), and used as host cells to generate Adv carrying apoptosis-inducing genes (proapoptotic genes). The 293-CrmA cells proved to be highly efficient for the construction of recombinant Adv carrying genes encoding Fas and Fas ligand. Moreover, the 293-CrmA line produced an ample quantity of these recombinant viruses. Because the conventional 293 plaque formation assay did not reflect the actual number of cells infected with the Adv carrying the proapoptotic gene, a determination of the Adv DNA copy number introduced into target cells was necessary to evaluate the quantity of infective virus. The techniques described here should be widely applicable for the construction of a recombinant Adv, in ample quantity, and for the estimation of the quantity of recombinant Adv produced. PMID- 9874267 TI - A novel strategy of cell targeting based on tissue-specific expression of the ecotropic retrovirus receptor gene. AB - Gene transfer into specific tissues or cell types is a key technique in the development of gene therapy. Modification of vector particles such that they selectively bind to the target cells has been attempted, but the limitation of this approach is the low transduction efficiency. Here, we show that a two-step gene transfer system can be used for efficient cell targeting. With this strategy, and using a high-titer adenoviral vector containing a tissue-specific promoter, we have engineered a system in which only target cells become susceptible to retrovirus-mediated transduction. In a model experiment, we constructed an adenoviral vector (Ad.AFPEcoRec) containing the ecotropic retrovirus receptor (EcoRec) gene under the control of the alpha-fetoprotein (AFP) promoter. A binding assay showed that after transduction with AD.AFPEcoRec, EcoRec molecules were efficiently expressed in AFP+HepG2 cells, but not in AFP HeLa and AFP-HLE cells. The EcoRec-expressing HepG2 cells could be stably transduced with ecotropic retroviral vectors, whereas HeLa and HLE cells remained highly resistant to retrovirus-mediated gene transfer. The apparent titer on HepG2 cells was greater than 2 x 10(5) CFU/ml. Because various tissue-specific promoter/enhancer elements are available, the two-step system could be used as a general strategy for both ex vivo and in vivo targeted gene transfer. PMID- 9874268 TI - Therapy of human ovarian cancer by transfection with the murine interferon beta gene: role of macrophage-inducible nitric oxide synthase. AB - The purpose of this study was to determine whether the local sustained production of murine interferon beta (mIFN-beta) could inhibit the growth of human ovarian cancer cells in the peritoneal cavity of nude mice. Human ovarian tumor Hey-A8 cells transfected with mIFN-beta (Hey-beta) or a control neomycin resistance vector (Hey-Neo) grew well in culture. Tumor cells were injected into the peritoneal cavity or under the subcutis of nuce mice. Parental (wild-type) or control transfected cells produced large tumors, whereas mIFN-beta-transfected cells did not produce any tumors. The IFN-beta-transfected cells prevented the outgrowth of bystander parental, control-transfected cells, and another human ovarian tumor cell line, SKOV3i.p.1, in the peritoneal cavity of nude mice. The IFN-beta-transfected tumor cells stimulated a high level of nitric oxide (NO) production in murine macrophages under both in vitro and in vivo conditions, and only the NO-producing macrophages exhibited antitumor activity. Collectively, these results demonstrate that the local production of IFN-beta can inhibit the in vivo growth of human ovarian cancer cells by upregulating the expression of the inducible nitric oxide synthase (iNOS) gene in host macrophages. PMID- 9874269 TI - High doses of a helper-dependent adenoviral vector yield supraphysiological levels of alpha1-antitrypsin with negligible toxicity. AB - Optimal gene therapy for many disorders will require efficient transfer to cells in vivo, high-level and long-term expression, and tissue-specific regulation, all in the absence of significant toxicity or inflammatory responses. While recombinant adenoviral vectors are efficient for gene transfer to hepatocytes, their usefulness is limited by short duration of expression related, at least in part, to immune responses to viral proteins and by a low capacity for foreign DNA. A number of systems have been developed for producing adenoviral vectors devoid of all viral coding sequences. Using AdSTK109, a vector lacking all viral coding sequences and carrying the complete human alpha1-antitrypsin (hAAT) genomic DNA locus, we have demonstrated sustained expression for longer than 10 months in mice. Utilizing high doses of this vector for hepatic gene transfer in mice, we find that supraphysiological levels of hAAT can be achieved without hepatotoxicity. PMID- 9874270 TI - Lentivirus-mediated transduction of islet grafts with interleukin 4 results in sustained gene expression and protection from insulitis. AB - Autoimmune destruction of islets in the pancreas leads to the development of insulin-dependent diabetes mellitus (IDDM). Replacement of insulin-producing tissue by transplantation of islets provides a cure to disease but requires immunosuppression or a means of controlling anti-graft immune responses. To promote islet survival we have utilized a local approach by expressing immunoregulatory molecules in islet grafts. The results presented here show that the human immunodeficiency virus (HIV)-based lentiviral vector is capable of stably transducing whole islets. Foreign reporter gene expression was observed both in vitro and in vivo 30 days after transplantation. Grafts containing insulin-positive beta-islet cells expressing foreign protein indicate that transduction does not interfere with glucose regulation. The absence of inflammatory infiltrates in grafts suggests that transduction does not activate the immune system. When islets transduced with an HIV vector expressing IL-4 were transplanted into diabetes-prone mice, animals were protected from autoimmune insulitis and islet destruction. As demonstrated by proliferative and cytokine analysis, protection was consistent with a switching of islet-antigen-specific T cell responses toward a Th2 phenotype. These results suggest that HIV-based lentivirus vectors can efficiently transduce islet cells with genes encoding potentially therapeutic molecules, for possibly managing diabetes. PMID- 9874271 TI - Long-term genetic modification of rhesus monkey hematopoietic cells following transplantation of adenoassociated virus vector-transduced CD34+ cells. AB - We have explored the potential of recombinant adenoassociated virus (AAV) vectors for gene transfer of the human beta-globin gene and the genetic modification of primate pluripotent hematopoietic stem cells (P-PHSCs). Transduction of P-PHSCs was tested in a preclinical bone marrow transplantation model in rhesus monkeys. CD34+ cells were transduced ex vivo and autologously transplanted without prior selection into irradiated rhesus monkeys. Vector-transduced peripheral blood mononuclear cells and granulocytes were present in the circulation for more than 15 months after transplantation. Approximately 1 in 10(5) cells in the circulation was vector modified. The vector was detected in the bone marrow, in granulocytes, and in purified populations of B and T cells, thus demonstrating multilineage repopulation by vector-transduced stem cells. Comparison of transduction protocols suggested that short-term culture of P-PHSCs enhances transduction and subsequent repopulation by rAAV-transduced cells. These results demonstrate that rAAV vectors can be used for the permanent genetic modification of a rhesus monkey hematopoietic system in the absence of selective pressure. PMID- 9874272 TI - Molecular lysis of synovial lining cells by in vivo herpes simplex virus thymidine kinase gene transfer. AB - Herpes simples virus thymidine kinase (HSV-TK) expression plasmid DNA was injected into the joint space of rabbits with antigen-induced arthritis (AIA). Purified plasmid DNA was able to mediate transfection of synovial lining cells and transient overexpression of HSV-TK in the context of active synovial inflammation. The pharmacodynamic distribution of intraarticular expression plasmid DNA was confined to the joint space. Arthritic rabbits treated with intraarticular expression plasmid DNA followed by intravenous ganciclovir (GCV, 5 mg/kg) twice daily for 3 days showed histologic evidence of synovial lining layer cytolysis when articular tissues were examined 21 days posttreatment. There was also a reduction in joint swelling in the TK-treated knees. No untoward clinical effects were observed in the rabbits and no evidence of cytolytic damage specific to the TK-GCV gene therapy was observed either in the articular cartilage or bone. The application of TK-GCV intraarticular gene therapy using purified expression plasmid DNA for the induction of synovial cytolysis may be applicable to the treatment of human inflammatory arthritis. PMID- 9874273 TI - Novel tools for production and purification of recombinant adenoassociated virus vectors. AB - Standard protocols for the generation of adenoassociated virus type 2 (AAV-2) based vectors for human gene therapy applications require cotransfection of cells with a recombinant AAV (rAAV) vector plasmid and a packaging plasmid that provides the AAV rep and cap genes. The transfected cells must also be overinfected with a helper virus, e.g., adenovirus (Ad), which delivers multiple helper functions necessary for rAAV production. Therefore, rAAV stocks produced using these protocols are contaminated with helper adenovirus. The generation of a novel packaging/helper plasmid, pDG, containing all AAV and Ad functions required for amplification and packaging of AAV vector plasmids, is described here. Cotransfection of cells with pDG and an AAV vector plasmid was sufficient for production of infectious rAAV, resulting in helper virus-free rAAV stocks. The rAAV titers obtained using pDG as packaging plasmid were up to 10-fold higher than those achieved using conventional protocols for rAAV production. Replacement of the AAV-2 p5 promoter by an MMTV-LTR promoter in pDG led to reduced expression of Rep78/68; however, expression of the VP proteins was significantly increased compared with VP levels from standard packaging plasmids. Immunofluorescence analyses showed that the strong accumulation of VP proteins in pDG-transfected cells resulted in enhanced AAV capsid assembly, which is limiting for efficient rAAV production. Furthermore, using a monoclonal antibody highly specific for AAV 2 capsids (A20), an rAAV affinity purification procedure protocol was established. The application of the tools described here led to a significant improvement in recombinant AAV vector production and purification. PMID- 9874274 TI - Polarity influences the efficiency of recombinant adenoassociated virus infection in differentiated airway epithelia. AB - To better understand mechanisms that limit rAAV transduction in the lung, we have evaluated several unique features of rAAV infection in polarized primary airway epithelial cultures. rAAV was found to transduce the basolateral surface of airway epithelia 200-fold more efficiently than the apical membrane. These differences in membrane infection correlated with the abundance of apical heparan sulfate proteoglycan (AAV-2 receptor) and virus binding. UV irradiation augmented rAAV transduction greater than 20-fold, only when virus was applied to the apical membrane. Ultrastructural analysis of UV-irradiated primary cultures demonstrated significant changes in microvilli architecture following exposure to 25 J/m2 UV. Although virus binding and the abundance of heparan sulfate proteoglycan were not increased at the apical membrane following UV irradiation, increased receptor independent endocytosis of fluorescent beads was seen at the apical membrane following UV irradiation. We hypothesize that endocytotic processes associated with apical membrane-specific pathways of viral entry, and/or processing of virus to the nucleus, may be altered following UV irradiation. Interestingly, UV irradiation had an inhibitory effect on rAAV transduction from the basolateral membrane, which correlated with a decrease in the abundance of heparan sulfate proteoglycan at the basal membrane. In summary, these findings suggest that independent pathways of viral transduction may occur in the apical and basolateral compartments of polarized airway epithelia. PMID- 9874275 TI - Adenoassociated virus-mediated transfer of a functional water channel into salivary epithelial cells in vitro and in vivo. AB - Aquaporin 1 (AQP1) is the archetypal member of a family of integral membrane proteins that function as water channels. Previously we have shown that this protein can be expressed transiently from a recombinant adenovirus (AdhAQP1) in vitro in different epithelial cell lines, and in vivo in rat submandibular glands. In the present study we have constructed a recombinant adenoassociated virus (rAAV) containing the human aquaporin 1 gene (rAAVhAQP1). rAAVhAQP1 was produced at relatively high titers. Approximately 10(11)-10(12) particles/ml and approximately 10(8)-10(9) transducing units/ml. We show that the rAAVhAQP1 can transduce in vitro four epithelial cell lines of different origins, at a level sufficient to detect the recombinant hAQP1 protein by either Western blot or confocal microscopic analysis. The recombinant hAQP1 was correctly targeted to the plasma membranes in all cell lines. Function of the recombinant hAQP1 was measured as fluid flow, in response to an osmotic gradient, across a monolayer of transduced epithelial cells. The data show that even at a low level of transduction, typically approximately 10% of the cells in the monolayer, transepithelial fluid movement is enhanced about threefold above basal levels. In addition, we report that rAAVhAQP1 can transduce epithelial cells in the salivary glands and liver of mice in vivo. These results suggest that rAAVs may be useful gene transfer vectors to direct the production of functional transgenes in salivary epithelial cell types. PMID- 9874276 TI - Herpes simplex virus type 1 DNA amplified as bacterial artificial chromosome in Escherichia coli: rescue of replication-competent virus progeny and packaging of amplicon vectors. AB - Herpes simplex virus type 1 (HSV-1)-based amplicon vectors contain only approximately 1% of the 152-kb HSV-1 genome, and consequently, replication and packaging into virions depends on helper functions. These helper functions have been provided conventionally by a helper virus, usually a replication-defective mutant of HSV-1, or more recently, by a set of five cosmids that overlap and represent the genome of HSV-1 deleted for DNA cleavage/packaging signals (pac). In the absence of pac signals, potential HSV-1 genomes that are reconstituted from the cosmids via homologous recombination are not packageable. The resulting amplicon stocks are, therefore, virtually free of contaminating helper virus. To simplify this packing system, the HSV-1 genome was cloned and maintained stably as a single-copy, F plasmid-based bacterial artificial chromosome in E. coli. Such a plasmid containing the HSV-1 genome deleted for the pac signals (fHSV delta pac) did not generate replication-competent progeny virus on transfection into mammalian cells, but rather, it was able to support the packaging of cotransfected amplicon DNA that contained a functional pac signal. The resulting amplicon vector stocks had titers of up to 10(7) transducing units per milliliter of culture medium and efficiently transduced neural cells in the rat brain, as well as hepatocytes in the rat. The capacity of generating infectious and replication-competent HSV-1 progeny following transfection into mammalian cells was restored after insertion of a pac signal into fHSV delta pac. PMID- 9874278 TI - Human gene marker/therapy clinical protocols. PMID- 9874277 TI - Thrombus generation after adenovirus-mediated gene transfer into atherosclerotic arteries. AB - Thrombosis represents a major issue during arterial local delivery. We evaluated the occurrence of thrombosis after adenovirus (Ad)-mediated gene transfer into normal and atherosclerotic arteries. A replication-deficient Ad vector expressing the beta-galactosidase reporter gene (Ad.RSV betagal; 4 x 10(9) PFU) was injected into normal and atherosclerotic arteries (n = 11 in both groups). The contralateral artery received either an Ad vector carrying no transgene (Ad.MLPnull) (n = 7 in both groups, 4 x 10(9) PFU) or vehicle buffer (n = 4 in normal group, n = 8 in atherosclerotic group). Animals were sacrificed 3 days following gene transfer for thrombus detection and assessment of beta galactosidase activity. Thrombus was absent in normal arteries and in atherosclerotic arteries injected with vehicle buffer only. In contrast, nonocclusive thrombus was present in atherosclerotic arteries injected with either Ad.RSV betagal (5 of 11) or Ad.MLPnull (3 of 7). Beta-galactosidase activity was predominantly found in the endothelial layer of the transfected arteries. Gene transfer and expression occurred despite the presence of the thrombus (4 of 5), and its efficiency did not significantly differ regardless of the thrombus. We conclude that thrombus frequently occurred in atherosclerotic arteries after Ad-mediated gene transfer. Further studies are warranted to identify the mechanisms of thrombus generation after Ad-mediated gene transfer into atherosclerotic arteries. PMID- 9874279 TI - Degradation of cytokeratin intermediate filaments by calcium-activated proteases (calpains) in vitro: implications for formation of Mallory bodies. AB - The degradation of rat hepatic intermediate filament (IF) proteins cytokeratin A (CK-A, 55-kDa) and cytokeratin D (CK-D, 48-kDa) by purified rat liver calcium activated proteases (calpains I and II) was evaluated in vitro. Calpain-mediated IF proteolysis was monitored by SDS-PAGE and Western blotting with antibodies to CK-A and CK-D and compared to microtubule protein actin. Both cytokeratins underwent rapid yet limited proteolysis by calpain I and II. Despite the conserved nature of cytokeratins and limited substrate specificity for calpains, distinct fragmentation patterns were obtained for calpain I) CK-A, 46- and 43 kDa/CK-D, 41-, and 39-kDa; and calpain II) CK-A, 46- and 43-kDa/CK-D 41-kDa. The 46-kDa CK-A fragment was the predominant fragment for both calpains. Two dimensional electrophoresis (IEF/SDS-PAGE) of CK fragments revealed the presence of classic "staircase" patterns consistent with endogenous proteases. Furthermore, proteolytic fragments showed a 2-D electrophoretic shift to lower pI suggesting that the limited hydrolysis occurred within the N-terminal arginine rich region of CK, a region believed essential for IF interactions in vivo. Thus, calpains may represent an initial step in the turnover of these stable and long lived proteins and as such, may be relevant to diseases characterized by abnormal disruption and bundling of IF such as formation of Mallory bodies in alcoholic hepatitis. PMID- 9874280 TI - Fetal copper uptake and a homolog (Atp7b) of the Wilson's disease gene in rats. AB - LEC rats are defective at the p-type copper transport ATPase gene (Atp7b) and exhibit excessive hepatic copper accumulation. However, copper concentration in fetal liver of LEC rats is lower than that of F344 normal rats. In this study, we made fetal backcrosses between LEC and F344 normal rats. At 19 days of gestation, hepatic copper concentrations of (LECXF344)F1XLEC and LECX(LECXF344)F1 fetuses were equivalent to those of F344 and LEC fetuses, respectively, irrespective of their Atp7b genotype. Furthermore, Atp7b expression was identified in the uterus and the maternal portion of placenta, but not in the fetal portion of placenta, in pregnant F344 rats. From these results, we speculate that the Atp7b product might contribute to a copper transport system from mother to fetus in the maternal portion, but not in the fetal portion of placenta. PMID- 9874281 TI - Effect of clarithromycin on alpha1-acid glycoprotein levels in normal and diabetic rats. AB - The elevation of alpha1-acid glycoprotein (AGP) serum level, a member of the acute phase proteins (APP) synthesized by the liver, is induced during inflammation. In this study, we show that clarithromycin (CAM), a 14-membered macrolide antibiotic, causes an increase in the serum levels of AGP in a dose dependent manner. AGP levels peak at 48h after a single administration with CAM In order to elucidate the mechanism of AGP induction by CAM, the effects of CAM, phenobarbital (PB), ethynylestradiol (EE2) and dexamethasone (DEX) treatments on APP fluctuation patterns were examined. In addition, modulation of the AGP induction by CAM, PB, EE2 and DEX in the diabetic state was examined. In contrast to treatment by PB, CAM treatment increased alpha2-macroglobulin levels to a much lesser extent than that observed during inflammation. On the other hand, albumin levels were unaffected by CAM treatment but decreased during inflammation. In addition, AGP induction by CAM, EE2 and DEX were attenuated in streptozotocin induced diabetic rats, whereas the PB-induced increase in AGP levels was potentiated in diabetic rats. Thus, CAM and DEX treatments appear to induce similar fluctuations in APP levels. These results suggest that CAM increases AGP levels via a mechanism which may be different from PB and typical inflammatory pathways. PMID- 9874282 TI - Mechanism of suppression of natural killer cell activity in trauma patients. AB - Trauma patients develop a severe immunosuppression that includes suppression of natural killer (NK) cell activity although numbers of NK cells are not reduced. The mechanism of suppression of NK cell activity after major trauma is not known. The aim of the present study was to investigate the in vitro effect of plasma samples from trauma patients (TP) on the cytotoxic activity of normal NK cells. Buffycoat mononuclear cells (5x10(5)/well) were preincubated with either TP or plasma samples from age and sex matched healthy controls (CP) for 0, 16 or 40 h. These effector cells were then cultured with 51Cr labeled K-562 cells (2x10(4)/well) for 4 h at 37 degrees C and % lysis was calculated. No significant differences in % lysis between CP and TP were found with 0 or 16 h preincubation, however 40 h preincubation with TP severely suppressed NK cell function (p=0.003) as compared to preincubation with CP for the same period. Addition of neutralizing anti-IL-4, anti-TGF-beta1, or anti-IL-10 antibodies did not reverse the NK cell suppression. There was a partial reversal of NK cell suppression by catalase but not by SOD or L-NMMA. Removal of monocytes from buffycoat mononuclear cells also significantly reversed the NK cell suppression. These data suggest that suppression of NK cell activity in trauma patients may be an accessory cell dependent phenomenon and may partially depend on production of reactive oxygen metabolites (ROM). PMID- 9874283 TI - Induction of oxidative stress and DNA damage by chronic administration of naphthalene to rats. AB - Naphthalene is a bicyclic aromatic compound that is widely used in various domestic and commercial applications including lavatory scent disks, soil fumigants and moth balls. Little information is available regarding the mechanism of naphthalene toxicity. We have assessed the oral, low dose (0.05 LD50) chronic effects of naphthalene (110 mg/kg/day p.o. in corn oil) for 120 consecutive days on lipid peroxidation and DNA fragmentation in the liver and brain tissues of female Sprague-Dawley rats. The animals were sacrificed on 0, 15, 30, 45, 60, 75, 90, 105 and 120 days of treatment. Maximum increases in hepatic and brain lipid peroxidation and DNA fragmentation were observed between 90 and 105 days of treatment. Following administration of naphthalene for 90 days, approximately 1.4 and 1.3-fold increases in lipid peroxidation were observed in the hepatic and brain tissues, respectively, while under the same conditions and time points 1.9- and 2.5-fold increases in hepatic and brain DNA fragmentation were observed, respectively. These results demonstrate that low dose chronic administration of naphthalene induces an oxidative stress resulting in tissue damaging effects that may contribute to the toxicity and carcinogenicity of naphthalene. PMID- 9874284 TI - Protective ability of acetylsalicylic acid (aspirin) to scavenge radiation induced free radicals in J774A.1 macrophage cells. AB - Radiation generates a variety of free radicals during the exposure of biological tissues through radiolysis of water. These free radicals are highly reactive and cause oxidative damage to biological molecules. This study examined the protective ability of aspirin against radiation induced oxidative stress. The study assessed the protective effect of aspirin (0.05 mM, 0.10 mM, 0.50 mM) on the generation of free radicals during exposure of J774A.1 macrophage cells to radiation (13.25 cGy). Approximately a 2.2-fold increase in superoxide anion formation as determined by cytochrome c reduction was observed following exposure of the cells to radiation for 20 one second exposures. Preincubation with aspirin exhibited a dose dependent decrease in free radical production as assessed by chemiluminescence and cytochrome c reduction. Aspirin also produced a concentration dependent reduction in radiation induced DNA damage in the cells. The data indicate that radiation of these cells results in production of reactive oxygen species and DNA damage, and aspirin can decrease these effects in a concentration dependent manner. PMID- 9874285 TI - Blood sampling site-dependent plasma concentrations and hepatotoxic parameters in serum after intraperitoneal administration of acetaminophen to rats. AB - Blood sampling site-dependent (the dorsal aorta, the jugular vein, and the retro orbital sinus) plasma concentrations of acetaminophen and its glucuronide and sulfate conjugates, and acetaminophen-induced hepatotoxic parameters (such as ALT and SDH activity) in serum were evaluated after intraperitoneal administration of acetaminophen, 500 mg/kg body weight, to rats. The plasma concentrations and the resultant AUC0-12 h of acetaminophen, acetaminophen-glucuronide, and acetaminophen-sulfate were significantly higher when blood samples were collected from the retro-orbital sinus than those from the jugular vein. The serum ALT activity at 3 and 24 h after administration of acetaminophen were significantly higher when the blood samples were collected from the retro-orbital sinus than those from the dorsal aorta. PMID- 9874286 TI - Radioprotective effects of 2-(allylthio)pyrazine an experimental chemopreventive agent: effects on detoxifying enzyme induction. AB - 2-(Allylthio)pyrazine (2-AP), which is effective in suppressing constitutive and inducible cytochrome P450 2E1 expression, exhibits hepatoprotective and chemopreventive effects. The radioprotective effects of 2-AP were examined in animals in association with the expression of microsomal epoxide hydrolase (mEH) and glutathione S-transferases (GSTs). 2-AP pretreatment (100 mg/kg/day, for 2 days) prior to total body irradiation (TBI) at the dose of 8 Gy increased the 30 day survival rate of mice to 91% from 48% in TBI alone. 2-AP caused an increase in the mean survival time of mice exposed to 9 Gy of TBI. Light microscopic examinations revealed that exposure of mice to 8 Gy of gamma-ray radiation resulted in hepatocyte degeneration in the surviving animals at day 1 through day 22 with certain extents of necrosis observed at early times, whereas 2-AP pretreatment protected the liver against ionizing radiation with no hepatic necrosis being observed. Mice irradiated at the dose of 8 Gy showed time-related decreases in the counts of WBC, RBC and platelet. 2-AP treatment, however, failed to protect the peripheral blood cells against gamma-irradiation and resulted in no improvement in the ratio of myeloid to erythroid bone marrow cells, as compared to TBI alone. Northern blot analysis revealed that exposure of mice to 8 Gy of TBI plus 2-AP exhibited greater mEH and mGSTA3 mRNA levels in the liver than those with TBI alone, although mGSTP1 mRNA level failed to be altered. Studies were also extended to determine the effects of 0.5 Gy gamma-irradiation in combination with 2-AP on the expression of hepatic mEH and GST genes in rats. Whereas mEH, rGSTA2, rGSTA3 and rGSTA5 mRNA levels were elevated 2- to 2.8-fold at 24 h after 2-AP treatment at the dose of 30 mg/kg, rats exposed to both 2-AP and 0.5 Gy of irradiation showed greater relative increases in the mRNAs. 2-AP enhanced the mEH and rGSTA2 gene expression to greater extents at day 1 after irradiation than after 3-5 consecutive daily treatment. The radiation-inducible mRNA levels of rGSTA3/5 and rGSTM1/2 were affected less by 2-AP pretreatment than were those of mEH and rGSTA2. These results demonstrate that 2-AP exhibits radioprotective efficacy against gamma-ray ionizing radiation in both mice and rats, which might be associated with enhanced expression of mEH and GST genes, but not with hematological improvement. PMID- 9874288 TI - A new method for the detection of bcr/abl sequences amplified by polymerase chain reaction (PCR) following bone marrow transplantation. AB - Disease recurrence is a major problem in patients receiving bone marrow transplantation for chronic myeloid leukemia. Residual malignant cells are the most likely source of recurrence. Detection of minimal residual disease early during therapy may provide an additional prognostic value and help in identifying patients who are at high risk of relapse. PCR followed by hybridization is the most sensitive method to investigate the persistence of leukemic cells. However, more reproducible methods suitable to standardization and quantification are required in clinical practice. In this study, we describe a novel PCR assay combined with immunological and colorimetric detection of the bcr-rearrangement. Residual bcr/abl rearranged cells were observed in 7 patients. Our results show that the assay is equally sensitive as RT-PCR, more versatile in terms of standardization and easily adaptable as a diagnostic test. PMID- 9874287 TI - Short-term macrophage interleukin-1 response to injected silicone in a rat model. AB - The short-term effects of silicone particles on the ability of splenic and peritoneal macrophages to produce Interleukin-1 (IL-1) were assessed. Lewis rats were divided into three groups. Group 1 animals (n=12) were injected subcutaneously with 2.5 ml of sterile saline; Group 2 animals (n=12) received 2.5 ml of Freund's Complete Adjuvant (FCA); Group 3 animals (n=12) received subcutaneous injections (2.5 ml) of a sonicated slurry comprised of equal parts of FCA and silicone. At days 1, 4, and 7 single-cell suspensions of splenic (10(8) cells/ml) and peritoneal (10(6) cells/ml) macrophages of four animals from each group were prepared. The supernatants from macrophages cultured in the presence or absence of lipopolysaccharide (LPS) were added to IL-1 dependent mouse thymocytes for 72 hrs, and subsequently pulsed with 3H-thymidine. Radioactivity incorporation was determined after 18 hrs. Peritoneal macrophage culture supernatants had significantly higher activity than splenic macrophages and all macrophages stimulated with LPS had significantly higher activity. At days 1 and 7, there were no significant differences in 3H-thymidine uptake. At day 4 there was an elevation of 3H-thymidine uptake from supernatant of peritoneal macrophages treated with FCA/silicone suggesting subcutaneous silicone injection has short-term effects. PMID- 9874289 TI - Changes in the weight of the thymus after birth and in pregnancy in mice. AB - The changes in the weight of thymus with age (days after birth) and effects of pregnancy on the weight of thymus were examined using mice. The weight of thymus increased after birth, and reached the maximum (107.5 mg) at the age of 40 days. It started to decrease, and became 48.9 mg at the age of 100 days. Thus, age related involution of the thymus in the female mice had started 50 days after birth. The weight of thymus was markedly reduced by pregnancy. The weight of thymus at the age of 60 days was 74.8 mg in the intact mice and 19.7 mg in the pregnant mice. Pregnancy greatly affects the weight of the thymus. PMID- 9874290 TI - Image guidance of therapy. PMID- 9874291 TI - Automated atlas integration and interactive three-dimensional visualization tools for planning and guidance in functional neurosurgery. AB - Many critical functionally distinct subcortical structures are not distinguishable on anatomical magnetic resonance imaging (MRI) scans. In order to provide the neurosurgeon with this missing information, a deformable volumetric atlas of the basal ganglia and thalamus has been created from the Schaltenbrand and Wahren atlas of cryogenic slices. The volumetric atlas can be automatically deformed to an individual patient's MRI. To facilitate the clinical use of the atlas, a visualization platform has been developed for preoperative and intraoperative use which permits manipulation of the merged atlas and MRI data sets in two- and three-dimensional views. The platform includes graphical tools which allow the visualization of projections of a leukotome and other surgical tools with respect to the atlas data, as well as preregistered images from any other imaging modality. In addition, a graphical interface has been designed to create custom virtual lesions using computer models of neurosurgical tools for intraoperative planning. To date this system has been employed as an adjunct to over 30 functional neurosurgical cases including surgery for movement disorders. PMID- 9874292 TI - Image guidance of breast cancer surgery using 3-D ultrasound images and augmented reality visualization. AB - This paper describes augmented reality visualization for the guidance of breast conservative cancer surgery using ultrasonic images acquired in the operating room just before surgical resection. By combining an optical three-dimensional (3 D) position sensor, the position and orientation of each ultrasonic cross section are precisely measured to reconstruct geometrically accurate 3-D tumor models from the acquired ultrasonic images. Similarly, the 3-D position and orientation of a video camera are obtained to integrate video and ultrasonic images in a geometrically accurate manner. Superimposing the 3-D tumor models onto live video images of the patient's breast enables the surgeon to perceive the exact 3-D position of the tumor, including irregular cancer invasions which cannot be perceived by touch, as if it were visible through the breast skin. Using the resultant visualization, the surgeon can determine the region for surgical resection in a more objective and accurate manner, thereby minimizing the risk of a relapse and maximizing breast conservation. The system was shown to be effective in experiments using phantom and clinical data. PMID- 9874293 TI - Predicting error in rigid-body point-based registration. AB - Guidance systems designed for neurosurgery, hip surgery, and spine surgery, and for approaches to other anatomy that is relatively rigid can use rigid-body transformations to accomplish image registration. These systems often rely on point-based registration to determine the transformation, and many such systems use attached fiducial markers to establish accurate fiducial points for the registration, the points being established by some fiducial localization process. Accuracy is important to these systems, as is knowledge of the level of that accuracy. An advantage of marker-based systems, particularly those in which the markers are bone-implanted, is that registration error depends only on the fiducial localization error (FLE) and is thus to a large extent independent of the particular object being registered. Thus, it should be possible to predict the clinical accuracy of marker-based systems on the basis of experimental measurements made with phantoms or previous patients. This paper presents two new expressions for estimating registration accuracy of such systems and points out a danger in using a traditional measure of registration accuracy. The new expressions represent fundamental theoretical results with regard to the relationship between localization error and registration error in rigid-body, point-based registration. Rigid-body, point-based registration is achieved by finding the rigid transformation that minimizes "fiducial registration error" (FRE), which is the root mean square distance between homologous fiducials after registration. Closed form solutions have been known since 1966. The expected value (FRE2) depends on the number N of fiducials and expected squared value of FLE, (FLE-2, but in 1979 it was shown that (FRE2) is approximately independent of the fiducial configuration C. The importance of this surprising result seems not yet to have been appreciated by the registration community: Poor registrations caused by poor fiducial configurations may appear to be good due to a small FRE value. A more critical and direct measure of registration error is the "target registration error" (TRE), which is the distance between homologous points other than the centroids of fiducials. Efforts to characterize its behavior have been made since 1989. Published numerical simulations have shown that (TRE2) is roughly proportional to (FLE2)/N and, unlike (FRE2), does depend in some way on C. Thus, FRE, which is often used as feedback to the surgeon using a point-based guidance system, is in fact an unreliable indicator of registration-accuracy. In this work we derive approximate expressions for (TRE2), and for the expected squared alignment error of an individual fiducial. We validate both approximations through numerical simulations. The former expression can be used to provide reliable feedback to the surgeon during surgery and to guide the placement of markers before surgery, or at least to warn the surgeon of potentially dangerous fiducial placements; the latter expression leads to a surprising conclusion: Expected registration accuracy (TRE) is worst near the fiducials that are most closely aligned! This revelation should be of particular concern to surgeons who may at present be relying on fiducial alignment as an indicator of the accuracy of their point-based guidance systems. PMID- 9874294 TI - Registration of real and CT-derived virtual bronchoscopic images to assist transbronchial biopsy. AB - This paper describes research work motivated by an innovative medical application: computer-assisted transbronchial biopsy. This project involves the registration, with no external localization device, of a preoperative three dimensional (3-D) computed tomography (CT) scan of the thoracic cavity (showing a tumor that requires a needle biopsy), and an intraoperative endoscopic two dimensional (2-D) image sequence, in order to provide assistance in transbronchial puncture of the tumor. Because of the specific difficulties resulting from the data being processed, a multilevel strategy was introduced. For each analysis level, the relevant information to process and the corresponding algorithms were defined. This multilevel strategy, thus, provides the best possible accuracy. Original image processing methods were elaborated, dealing with segmentation, registration and 3-D reconstruction of the bronchoscopic images. In particular, these methods involve adapted mathematical morphology tools, a "daemon-based" registration algorithm, and a model-based shape-from-shading algorithm. This pilot study presents the application of these algorithms to recorded bronchoscopic video sequences for five patients. The preliminary results presented here demonstrate that it is possible to precisely localize the endoscopic camera within the CT data coordinate system. The computer can thus synthesize in near real-time the CT-derived virtual view that corresponds to the actual endoscopic view. PMID- 9874295 TI - Anatomy-based registration of CT-scan and intraoperative X-ray images for guiding a surgical robot. AB - We describe new methods for rigid registration of a preoperative computed tomography (CT)-scan image to a set of intraoperative X-ray fluoroscopic images, for guiding a surgical robot to its trajectory planned from CT. Our goal is to perform the registration, i.e., compute a rotation and translation of one data set with respect to the other to within a prescribed accuracy, based upon bony anatomy only, without external fiducial markers. With respect to previous approaches, the following aspects are new: 1) we correct the geometric distortion in fluoroscopic images and calibrate them directly with respect to the robot by affixing to it a new calibration device designed as a radiolucent rod with embedded metallic markers, and by moving the device along two planes, while radiographs are being acquired at regular intervals; 2) the registration uses an algorithm for computing the best transformation between a set of lines in three space, the (intraoperative) X-ray paths, and a set of points on the surface of the bone (imaged preoperatively), in a statistically robust fashion, using the Cayley parameterization of a rotation; and 3) to find corresponding sets of points to the X-ray paths on the surfaces, our new approach consists of extracting the surface apparent contours for a given viewpoint, as a set of closed three-dimensional nonplanar curves, before registering the apparent contours to X-ray paths. Aside from algorithms, there are a number of major technical difficulties associated with engineering a clinically viable system using anatomy and image-based registration. To detect and solve them, we have so far conducted two experiments with the surgical robot in an operating room (OR), using CT and fluoroscopic image data of a cadaver bone, and attempting to faithfully simulate clinical conditions. Such experiments indicate that intraoperative X-ray-based registration is a promising alternative to marker based registration for clinical use with our proposed method. PMID- 9874296 TI - Treatment planning of brain implants using vascular information and a new template technique. AB - A new template technique has been developed for implanting hyperthermia catheters in the treatment of brain tumors. The technique utilizes an imaging template and a drill template which can be rigidly secured to the head with three skull screws. The anatomic and vascular information needed for hyperthermia treatment planning may be assessed with three-dimensional magnetic resonance (MR) imaging and angiography acquisitions which use a surface coil. In the companioning treatment planning system the catheter positions and lengths and the electrodes in the catheter can be interactively manipulated relative to the anatomy and vasculature. The visualization of the blood vessels relative to the template allows the minimization of the risk on intracranial hemorrhages. This template technique is useful for any brain tumor implants, especially when a large number of catheters are involved. A phantom test has shown that this procedure has an accuracy in the order of 1 mm provided that the MR-related geometry distortions are minimized. PMID- 9874297 TI - Calibration of tracking systems in a surgical environment. AB - The purpose of this paper was to assess to what extent an optical tracking system (OTS) used for position determination in computer-aided surgery (CAS) can be enhanced by combining it with a direct current (dc) driven electromagnetic tracking system (EMTS). The main advantage of the EMTS is the fact that it is not dependent on a free line-of-sight. Unfortunately, the accuracy of the EMTS is highly affected by nearby ferromagnetic materials. We have explored to what extent the influence of the metallic equipment in the operating room (OR) can be compensated by collecting precise information on the nonlinear local error in the EMTS by using the OTS for setting up a calibration look-up table. After calibration of the EMTS and registration of the sensor systems in the OR we have found the average euclidean deviation in position readings between the dc tracker and the OTS reduced from 2.9+/-1.0 mm to 2.1+/-0.8 mm within a half-sphere of 530 mm radius around the magnetic field emitter. Furthermore we have found the calibration to be stable after re-registration of the sensors under varying conditions such as different heights of the OR table and varying positions of the OR equipment over a longer time interval. These results encourage the further development of a hybrid magnetooptical tracker for computer-aided surgery where the electromagnetic tracker acts as an auxiliary source of position information for the optical system. Strategies for enhancing the reliability of the proposed hybrid magnetooptic tracker by detecting artifacts induced by mobile ferromagnetic objects such as surgical tools are discussed. PMID- 9874298 TI - Surface-based registration of CT images to physical space for image-guided surgery of the spine: a sensitivity study. AB - This paper presents a method designed to register preoperative computed tomography (CT) images to vertebral surface points acquired intraoperatively from ultrasound (US) images or via a tracked probe. It also presents a comparison of the registration accuracy achievable with surface points acquired from the entire posterior surface of the vertebra to the accuracy achievable with points acquired only from the spinous process and central laminar regions. Using a marker-based method as a reference, this work shows that submillimetric registration accuracy can be obtained even when a small portion of the posterior vertebral surface is used for registration. It also shows that when selected surface patches are used, CT slice thickness is not a critical parameter in the registration process. Furthermore, the paper includes qualitative results of registering vertebral surface points in US images to multiple CT slices. The method has been tested with US points and physical points on a plastic spine phantom and with simulated data on a patient CT scan. PMID- 9874299 TI - Registration of head CT images to physical space using a weighted combination of points and surfaces. AB - Most previously reported registration techniques that align three-dimensional image volumes by matching geometrical features such as points or surfaces use a single type of feature. We recently reported a hybrid registration technique that uses a weighted combination of multiple geometrical feature shapes. In this study we use the weighted geometrical feature (WGF) algorithm to register computed tomography (CT) images of the head to physical space using the skin surface only, the bone surface only, and various weighted combinations of these surfaces and one fiducial point (centroid of a bone-implanted marker). We use data acquired from 12 patients that underwent temporal lobe craniotomies for the resection of cerebral lesions. We evaluate and compare the accuracy of the registrations obtained using these various approaches by using as a reference gold standard the registration obtained using three bone-implanted markers. The results demonstrate that a combination of geometrical features can improve the accuracy of CT-to physical space registration. Point-based registration requires a minimum of three noncolinear points. The position of a bone-implanted marker can be determined much more accurately than that of a skin-affixed marker or an anatomic landmark. A major disadvantage of using bone-implanted markers is that an invasive procedure is required to implant each marker. By combining surface information, the WGF algorithm allows registration to be performed using only one or two such markers. One important finding is that the use of a single very accurate point (a bone-implanted marker) allows very accurate surface-based registration to be achieved using very few surface points. Finally, the WGF algorithm, which not only allows the combination of multiple types of geometrical information but also handles point-based and surface-based registration as degenerate cases, could form the foundation of a "flexible" surgical navigation system that allows the surgeon to use what he considers the method most appropriate for an individual clinical situation. PMID- 9874300 TI - Pubic arch detection in transrectal ultrasound guided prostate cancer therapy. AB - New biopsy techniques, increased life expectancy, and prostate-specific antigen (PSA) screening have contributed to an increase in the reported incidence of prostate cancer. Among several treatment options available to the patients, transperineal prostate brachytherapy has emerged as a medically successful, cost effective outpatient procedure for treating localized prostate cancer. Transperineal prostate brachytherapy employs transrectal ultrasound (TRUS) as the primary imaging modality to accurately preplan and subsequently execute the placement of radioactive seeds into the prostate. Under TRUS guidance, a needle (preloaded with radioactive seeds) is inserted through a template guide, through the perineum and into a predetermined prostate target. The pubic arch, formed by the central union of pelvic bones, is a potential barrier to the passage of these needles in the prostate. A critical aspect, therefore, in the planning and execution of the brachytherapy procedure is the accurate assessment of pubic arch interference (PAI) in relation to the prostate. Traditionally, the evaluation of PAI has involved computed tomography correlate scanning or crude subjective evaluations. In this paper, we describe a new method of assessing PAI by detecting the pubic arch via image processing on the TRUS images. The PAI detection (PAID) algorithm first uses a technique known as sticks to selectively enhance the contrast of linear features in ultrasound images. Next, the enhanced image is thresholded via percentile thresholding. Finally, we fit a parabola (a model for the pubic arch) recursively to the thresholded image. Our evaluation result from 15 cases indicates that the algorithm can successfully detect the pubic arch with 90% accuracy. Based on this study, we believe that detecting the pubic arch and assessing PAI can be done practically and more accurately in the clinical setting using TRUS rather than the current available methods. PMID- 9874301 TI - Guidance of intracoronary radiation therapy based on dose-volume histograms derived from quantitative intravascular ultrasound. AB - Application of ionizing radiation to prevent restenosis in atherosclerotic vessels treated by balloon angioplasty is a new treatment under investigation in interventional cardiology and radiology. There is variability in dose prescription, and both gamma- and beta-emitters are used, leading to a wide range of dose distribution over the arterial vessel wall. We present a new modality of dosimetry based on a method that three-dimensional (3-D) image reconstruction of electrocardiogram (ECG)-gated intravascular ultrasound (IVUS) images. Dose volume histograms (DVH) are used to describe the cumulative distribution of dose over two specific volumes: i) at the level of the luminal surface, defined with a thickness of 0.1 mm from the automatically detected contour of the highly echogenic blood-vessel interface, and ii) the adventitia volume is computed considering a 0.5-mm thickness from the echogenic media-adventitia interface. DVH provide a tool for reporting the actual delivered dose at the site believed to be the target: the adventitia, and to detect excessive radiation which could lead to vascular complications. Simulation of a gamma-emitter or of a radioactive source train in the center of the lumen are possible. The data obtained from the first ten patients included in the beta-irradiation trial (BERT 1.5) conducted in our institution are presented, supporting the use of DVH based on quantitative IVUS measurements for optimal dose prescription in vascular interventional radiation therapy. PMID- 9874302 TI - Image guidance of endovascular interventions on a clinical MR scanner. AB - Magnetic resonance imaging (MRI) offers potential advantages over conventional X ray techniques for guiding and evaluating vascular interventions. Image guidance of such interventions via passive catheter tracking requires real-time image processing. Commercially available MR scanners currently do not provide this functionality. This paper describes an image processing environment that allows near-real-time MR-guided vascular interventions. It demonstrates 1) that flexibility can be achieved by separating the scanner and the image processing/display system, thereby preserving the stability of the scanner and 2) that sufficiently rapid visualization can be achieved by low-cost workstations equipped with graphics hardware. The setup of the hardware and the software is described in detail. Furthermore, image processing techniques are presented for guiding the interventionalist through simple vascular anatomy. Finally, results of a phantom balloon angioplasty experiment are presented. PMID- 9874303 TI - Device visualization for interventional MRI using local magnetic fields: basic theory and its application to catheter visualization. AB - This paper addresses one of the major problems in Interventional magnetic resonance imaging (MRI): the visualization of interventional devices. For visualization locally induced magnetic fields are used, which disturb the homogeneity of the main magnetic field of the MR scanner. This results in signal loss in the vicinity of the device due to intravoxel dephasing, and leads to a disturbance of the phase image. The local fields are established by a low current in a closed copper loop along the device. This method will be introduced as a means for catheter visualization. The basic theory behind this method is presented. Simulations are performed to determine the effect of intravoxel dephasing, without interfering effects like susceptibility or radio-frequency artifacts. Scanned and simulated data is used to verify the theoretical consideration. Different configurations of wire loops are discussed and two types of catheter visualization scans are proposed. Results from a pig study show that this methods holds promise for intravascular interventions under MRI guidance. PMID- 9874304 TI - Methods for providing probe position and temperature information on MR images during interventional procedures. AB - Interventional magnetic resonance imaging (MRI) can be defined as the use of MR images for guiding and monitoring interventional procedures (e.g., biopsy, drainage) or minimally invasive therapy (e.g., thermal ablation). This work describes the development of a prototype graphical user interface and the appropriate software methods to accurately overlay a representation of a rigid interventional device [e.g., biopsy needle, radio-frequency (RF) probe] onto an MR image given only the probe's spatial position and orientation as determined from a three-dimensional (3-D) localizer used for interactive scan plane definition. This permits 1) "virtual tip tracking," where the probe tip location is displayed on the image without the use of separate receiver coils or a "road map" image data set, and, 2) "extending" the probe to predict its path if it were directly moved forward toward the target tissue. Further, this paper describes the design and implementation of a method to facilitate the monitoring of thermal ablation procedures by displaying and overlaying temperature maps from temperature sensitive MR acquisitions. These methods provide rapid graphical updates of probe position and temperature changes to aid the physician during the actual interventional MRI procedures without altering the usual operation of the MR imager. PMID- 9874305 TI - Dynamic tracking in interventional MRI using wavelet-encoded gradient-echo sequences. AB - This work describes a newly developed magnetic resonance imaging (MRI) data acquisition strategy which replaces the standard Fourier phase-encoding with the spatially localized coefficients of wavelet-encoding and offers a new technique for image guidance when combined with a dynamic tracking algorithm. By using this new technique, only a specific fraction of the entire raw data set needs to be updated and reconstructed to visualize the movement of an interventional device during an MR guided procedure. The combination of wavelet-encoding and a dynamic tracking algorithm was implemented in two-dimensional and three-dimensional gradient-echo sequences on a 0.2-T open C-arm-shaped MR system (Siemens, Erlangen Germany) and tested in phantom and in vitro experiments. When applying the wavelet-encoding direction parallel to the movement of a straight interventional device, only those spatially localized wavelet-coefficients mainly affected by the interventional device are updated. This led to potential increases of the image frame rate by a factor of up to seven. PMID- 9874306 TI - Validation of object-induced MR distortion correction for frameless stereotactic neurosurgery. AB - Spatial fidelity is a paramount issue in image guided neurosurgery. Until recently, three-dimensional computed tomography (3D CT) has been the primary modality because it provides fast volume capture with pixel level (1 mm) accuracy. While three-dimensional magnetic resonance (3D MR) images provide superior anatomic information, published image capture protocols are time consuming and result in scanner- and object-induced magnetic field inhomogeneities which raise inaccuracy above pixel size. Using available scanner calibration software, a volumetric algorithm to correct for object-based geometric distortion, and a Fast Low Angle SHot (FLASH) 3D MR-scan protocol, we were able to reduce mean CT to MR skin-adhesed fiducial marker registration error from 1.36 to 1.09 mm. After dropping the worst one or two of six fiducial markers, mean registration error dropped to 0.62 mm (subpixel accuracy). Three dimensional object-induced error maps present highest 3D MR spatial infidelity at the tissue interfaces (skin/air, scalp/skull) where frameless stereotactic fiducial markers are commonly applied. The algorithm produced similar results in two patient 3D MR-scans. PMID- 9874307 TI - Investigation of intraoperative brain deformation using a 1.5-T interventional MR system: preliminary results. AB - All image-guided neurosurgical systems that we are aware of assume that the head and its contents behave as a rigid body. It is important to measure intraoperative brain deformation (brain shift) to provide some indication of the application accuracy of image-guided surgical systems, and also to provide data to develop and validate nonrigid registration algorithms to correct for such deformation. We are collecting data from patients undergoing neurosurgery in a high-field (1.5 T) interventional magnetic resonance (MR) scanner. High-contrast and high-resolution gradient-echo MR image volumes are collected immediately prior to surgery, during surgery, and at the end of surgery, with the patient intubated and lying on the operating table in the operative position. In this paper we report initial results from six patients: one freehand biopsy, one stereotactic functional procedure, and four resections. We investigate intraoperative brain deformation by examining threshold boundary overlays and difference images and by measuring ventricular volume. We also present preliminary results obtained using a nonrigid registration algorithm to quantify deformation. We found that some cases had much greater deformation than others, and also that, regardless of the procedure, there was very little deformation of the midline, the tentorium, the hemisphere contralateral to the procedure, and ipsilateral structures except those that are within 1 cm of the lesion or are gravitationally above the surgical site. PMID- 9874308 TI - True energy-minimal and finite-size biplanar gradient coil design for MRI. AB - Finite-sized high-performance planar magnetic field gradient coils in today's open configuration magnetic resonance imaging (MRI) systems have always been desirable for ever demanding imaging applications. We present a Lagrange multiplier technique for designing a minimum-energy gradient coil under a finite size planar geometry constraint in addition to a set of magnetic field constraints. In this new design methodology, the surface current density on a finite size plane is represented by a two-dimensional (2-D) Fourier series expansion. Following the standard approach, we construct a functional F in terms of the stored magnetic energy and a set of field constraint points which are chosen over the desired imaging volume. Minimizing F, we obtain the continuous current density distribution for the finite-size planar gradient coil. Applying the stream function technique to the resulting continuous current distribution, the discrete current pattern can be generated. Employing the Biot-Savart law to the discrete current loops, the gradient magnetic field has been re-evaluated in order to validate the theory. Using this approach, we have been able to design a finite-size biplanar z-gradient coil which is capable of generating a gradient field of 40 mT/m @ 266 A. The excellent agreement between the analytical and numerical results has been achieved. PMID- 9874309 TI - CTA-based angle selection for diagnostic and interventional angiography of saccular intracranial aneurysms. AB - Coil embolization is a safe treatment for cerebral aneurysms only if the width of the neck in relation to the fundus of the aneurysm is small. Therefore, accurate visualization of the aneurysmal neck is required both in the diagnostic process and during the intervention. Conventional digital subtraction angiography (DSA) is still the preferred modality for the examination of cerebrovascular abnormalities like aneurysms, but it often does not provide the required morphological characteristics due to the suboptimal selection of projection angles and resulting overprojections of surrounding vasculature. This paper presents a method for performing a computer-assisted calculation of the optimal projection angles for DSA by post-processing computed tomographic angiography (CTA) volume data using ray-casting techniques and a combination of image processing algorithms. By means of phantom studies, retrospective simulations of angiograms, and in vivo applications of calculated optimal viewing angles, it is demonstrated that the proposed method results in better angiographic projections of the neck of saccular aneurysms with small neck-fundus ratio than those acquired at standard angles prescribed by clinical protocols. PMID- 9874310 TI - An image-guided planning system for endosseous oral implants. AB - A preoperative planning system for oral implant surgery was developed which takes as input computed tomographies (CT's) of the jaws. Two-dimensional (2-D) reslices of these axial CT slices orthogonal to a curve following the jaw arch are computed and shown together with three-dimensional (3-D) surface rendered models of the bone and computer-aided design (CAD)-like implant models. A technique is developed for scanning and visualizing an eventual existing removable prosthesis together with the bone structures. Evaluation of the planning done with the system shows a difference between 2-D and 3-D planning methods. Validation studies measure the benefits of the 3-D approach by comparing plans made in 2-D mode only with those further adjusted using the full 3-D visualization capabilities of the system. The benefits of a 3-D approach are then evident where a prosthesis is involved in the planning. For the majority of the patients, clinically important adjustments and optimizations to the 2-D plans are made once the 3-D visualization is enabled, effectively resulting in a better plan. The alterations are related to bone quality and quantity (p < 0.05), biomechanics (p < 0.005), and esthetics (p < 0.005), and are so obvious that the 3-D plan stands out clearly (p < 0.005). The improvements often avoid complications such as mandibular nerve damage, sinus perforations, fenestrations, or dehiscences. PMID- 9874311 TI - An iterative algorithm for X-ray CT fluoroscopy. AB - X-ray computed tomography fluoroscopy (CTF) enables image guidance of interventions, synchronization of scanning with contrast bolus arrival, and motion analysis. However, filtered backprojection (FB), the current method for CTF image reconstruction, is subject to motion and metal artifacts from implants, needles, or other surgical instruments. Reduced target lesion conspicuity may result from increased image noise associated with reduced tube current. In this report, we adapt the row-action expectation-maximization (EM) algorithm for CTF. Because time-dependent variation in images is localized during CTF, the row action EM-like algorithm allows rapid convergence. More importantly, this iterative CTF algorithm has fewer metal artifacts and better low-contrast performance than FB. PMID- 9874312 TI - Non-compliance following renal transplantation in children and adolescents. AB - Reported frequencies of non-compliance in children with end-stage renal disease range from 8% to 70% with a mean around 40%. Sequelae amount to momentous emotional and financial burdens, including the loss of 7% of transplanted organs. Reasons for non-compliance have too often been attributed selectively to the patients (e.g., emotional, mental, social, or communication problems). Compared with general compliance research, this selective attribution appears to be too simplistic. Selective attribution neglects the patients' experiences within the context of disease and treatment and prevents open communication about non compliance. Research on personal reasons for non-compliance is scarce. In psychological interviews, a third of our 85 patients with end-stage renal disease (34 boys, 51 girls, mean age 12.7 years, range 7.4-19.3 years) communicated psychologically meaningful reasons for non-compliance, frequently related to interrelational and systemic treatment conditions. Patients indirectly asked for more communication about their subjective reasons for non-compliance. PMID- 9874313 TI - Kidney transplantation after a severe form of pseudotumor cerebri. AB - Pseudotumor cerebri is a syndrome characterized by intracranial hypertension (intracranial pressure >200 mmH2O) and a normal ventricular system. The diagnosis should be made as early as possible to prevent impairment of vision. Several diseases have been reported in association with pseudotumor cerebri in pediatric patients, and have been occasionally also noted with chronic renal failure, heart and renal transplantation. We report a 7-year-old boy who complained of severe headaches and visual impairment 2 years after hemodialysis for renal hypoplasia. Pseudotumor cerebri was suspected and, despite treatment with corticosteroids, acetazolamide, and lumboperitoneal diversion, visual impairment worsened. Bilateral optic nerve sheath decompression (ONSD) was performed without success and the child completely lost his vision within 2 weeks. He was successfully transplanted 2 months later. Two years post transplantation, the blind child has a normal renal function and school performance. Pseudotumor cerebri must be rapidly suspected in a child with renal failure suffering from headaches and papilledema. Visual loss may progress rapidly and ONSD seems to be the best surgical treatment when medical treatment fails. In this patient renal transplantation was well tolerated, with no deterioration in the neurological status over 2 years of follow-up. PMID- 9874314 TI - The role of PAX2 in normal and abnormal development of the urinary tract. AB - The molecular etiology of many urinary tract abnormalities in children remains unknown, but a number of genes with a key role in urogenital development have now been identified. PAX2, one such gene, encodes a transcription factor which is critically required for epithelial differentiation within the urogenital tract. Recent studies suggest that PAX2 mutations lead to urological abnormalities and renal failure, while overexpression of PAX2 in the kidneys of mice causes multifocal microcystic tubular dilatation. In humans persistent PAX2 expression has been identified in multicystic dysplastic kidneys. In this review, recent research on the developmental gene, PAX2, and its involvement in normal and abnormal kidney development is summarized. In addition, an overview of the phenotypes associated with either loss-of-function PAX2 mutations or PAX2 overexpression is presented. A brief summary of factors that are known to regulate PAX2 and genes that may be regulated by PAX2 protein is also included. PMID- 9874315 TI - The molecular biology of polycystic kidney disease. AB - In recent years there have been a number of developments in polycystic kidney disease (PKD) research. The genes associated with the predominant forms of autosomal dominant PKD have been cloned, and the gene associated with a mouse model for autosomal recessive PKD has been identified and characterized. Other studies have yielded new information regarding the role of the epidermal growth factor receptor gene in promoting renal cyst formation. In this review article we summarize recent published data on the molecular genetics of autosomal dominant and autosomal recessive PKD and provide a working model of how multiple genes participate in the PKD disease pathway. PMID- 9874316 TI - Childhood reflux and urinary infection: a follow-up of 10-41 years in 226 adults. AB - To ascertain the outcome of childhood vesicoureteric reflux (VUR), 226 adults (37 males), mean age 27 years, were studied after 10-35 years, extended to 41 years by postal questionnaire in 161. At presentation (mean age 5 years) all had VUR (grade III-V in 68) and urinary tract infection (UTI); there was renal scarring in 85 (acquired before referral in 11 and during follow-up in 1), hypertension in 6 and impaired renal function in 5. They were managed and followed prospectively by one paediatrician; 63% of these children remained free from UTI; VUR persisted in 63 and had resolved in 69% of 193 children managed medically. At follow-up, 61% of adults had remained free from infection; 17 adults had hypertension and/or raised plasma creatinine, 16 with scarred kidneys. Their deterioration was predictable because of scar type, blood pressure or plasma creatinine levels in childhood. No new scars developed after puberty. Renal growth rates were unaffected by initial severity or persistence of VUR. On the later questionnaire, 9 further adults, mean age 38 years, had moderate hypertension. The adults with complications were those with extensive renal scarring and/or at least borderline hypertension in childhood. Those with VUR, but no scarring, and managed carefully in childhood, did not suffer serious consequences as adults. There is a need for early recognition and treatment of children with VUR and UTI to limit scar development. PMID- 9874318 TI - Renal histology and response to cyclosporine in childhood idiopathic nephrotic syndrome. PMID- 9874317 TI - Quantitative morphometry of renal biopsies prior to cyclosporine in nephrotic syndrome. AB - Use of cyclosporine (CsA) in the management of children with steroid-resistant (SRNS) and steroid-dependent (SDNS) nephrotic syndrome has become increasingly popular in recent years. Although most children receive a renal biopsy prior to initiation of CsA, the relationship between initial renal histology and the subsequent clinical response to CsA is not known. We analyzed the correlation between pre-CsA segmental and global glomerular scarring and interstitial fibrosis and the subsequent response to CsA in 23 children (5.6+/-1.0 years, Mean+/-SEM) with SDNS (n=8) and SRNS (n=15) treated with CsA for 24.2+/-3.8 months and followed for 28.0+/-4.1 months. Complete remission was obtained in 78% of patients within 67.6+/-16 days, while 18% had a partial response and 4% no response. Quantitative histological analysis revealed a trend toward partial rather than complete response with increasing segmental glomerular (P=0.13), global glomerular (P=0.05), and interstitial (P=0.08) scarring, and among patients with minimal change nephrotic syndrome versus IgM nephropathy versus focal segmental glomerulosclerosis. Among complete responders, linear regression analyses revealed no correlation between time to response and pre-CsA glomerular or interstitial scarring. We conclude that increased glomerular or interstitial scarring on a pre-CsA renal biopsy tends to correlate with a partial, rather than complete, response to CsA in childhood nephrotic syndrome. PMID- 9874319 TI - Long-term effects of immunosuppressants in steroid-dependent nephrotic syndrome. AB - In order to elucidate long-term effects of immunosuppressants, we studied 60 children with steroid-dependent nephrotic syndrome who were treated with three immunosuppressants: cyclophosphamide (n=34), chlorambucil (n=11), and cyclosporin A (n=15). Each relapse before and after the administration of immunosuppressants was evaluated longitudinally in terms of the relapse-free period and the maintenance dose of prednisolone required. The median follow-up period after immunosuppressants was 5.2 years (range 0.5-20.3 years). The relapse-free period was significantly longer in all groups after the initiation of immunosuppressants. However, the relapse-free period after subsequent relapses as compared with the previous relapse was longer in the cyclophosphamide group, similar in the chlorambucil group, and shorter in the cyclosporin A group. The prednisolone dosage at relapse was reduced in subsequent relapses after cyclophosphamide and chlorambucil treatment, but tended to be higher in later relapses after the initiation of cyclosporin A. These findings suggest that the effects of cyclophosphamide are long lasting, while those of chlorambucil and cyclosporin A are of short duration. Children who relapse after cyclosporin A treatment may experience a worse relapsing course. PMID- 9874320 TI - Antioxidant status of children with steroid-sensitive nephrotic syndrome. AB - Eighteen children with steroid-sensitive nephrotic syndrome (SSNS) were studied. The control group comprised 20 healthy children. The following indirect parameters of reactive oxygen species activity were determined in nephrotic patients during four stages of the disease (full relapse before prednisone administration, disappearance of proteinuria, prednisone cessation, unmaintained remission): plasma malondialdehyde (MDA) levels, copper/zinc superoxide dismutase (CuZn SOD) activity and glutathione peroxidase (GPX) activity in erythrocytes, reduced glutathione (GSH) and vitamin C levels in whole blood, and vitamin E level in serum. Increased MDA levels, reduced vitamin C levels, and enhanced CuZn SOD activity were found in relapse. GSH concentration was high during all four stages. Vitamin E level was also increased, parallel to the pattern of serum lipids. GPX activity remained low during the proteinuria stage and in remission. We conclude that the majority of abnormal findings can be attributed to the hyperlipidemia of NS. Low GPX activity may be a factor limiting the antioxidant capacity in NS. The present study is inconclusive regarding the role of free radicals in the proteinuria of NS. PMID- 9874321 TI - Lymphocyte ectoenzymes in childhood idiopathic nephrotic syndrome. AB - Lymphocyte ectoenzymes with immunomodulatory function were investigated in 11 children with minimal change disease (MCD), 9 with primary focal segmental glomerulosclerosis (FSGS), and 17 age- and sex-matched healthy children. Basal, concanavalin A (Con A)-, and pokeweed mitogen (PWM)-stimulated lymphocyte ecto-5' nucleotidase (5'-Nu), dipeptidyl peptidase IV (DPP IV), and alkaline phosphodiesterase I (APD) activities were determined. In MCD relapse ecto-APD activity of unstimulated lymphocytes was higher than controls. Ecto-APD of Con A stimulated lymphocytes was below controls (23.0, range 7.2-48.7 nmol/min per 10(6) lymphocytes) in all active MCD (18.7, range 7.6-32.6), during corticosteroid treatment (14.6, range 4.5-54), and in remission (13.1, range 6.1 19.6), but was significant only in remission. Con A-stimulated DPP IV was significantly lower from controls (53.8, range 19.3-85.7 nmol/min per 10(6) lymphocytes) in all active MCD (38.1, range 10.8-82.1), during treatment (37.5, range 20.2-58.7), and in remission (39.4, range 24.3-69.6). In FSGS, unstimulated lymphocyte ecto-APD activity was greater than controls. However, Con A-stimulated lymphocyte ecto-APD and DPP IV activities were not significantly different from controls. Con A stimulation of lymphocyte ecto-APD and DPP IV activity was significantly reduced in MCD relapse and in remission, but not in FSGS. Basal, Con A-, and PWM-stimulated ecto-5'-Nu in MCD and FSGS were not different from controls. These results suggest a role for abnormal T cell function in MCD but not in FSGS. The difference in mitogen-stimulated expression of these ectoenzymes suggests a different pathogenesis of childhood MCD and primary FSGS. PMID- 9874322 TI - IgG subclass/IgM ratio and response to therapy in focal segmental glomerulonecrosis. AB - Focal segmental glomerulosclerosis (FSGS) is relatively steroid resistant and no clinical or histological marker can predict the response to therapy. To investigate the role of serum immunoglobulin subclass/IgM in predicting the response to therapy in FSGS, serum concentrations of total IgG, IgG subclasses, and the ratio of serum IgG subclasses to total IgG (% IgG subclass) were measured in 27 children during the acute nephrotic state. Prednisolone, cyclophosphamide, and Persantine (dipyridamole) were given for 12 weeks. We divided the patients into good responders or poor responders according to clinical response. The clinical and nephrotic status were similar in both groups. Fourteen patients were good responders with higher serum IgGI/IgM than that of non-responders (4.00+/ 0.67 vs. 1.61+/-0.20, P<0.05). There was no significant difference in IgG2/IgM between these two groups. These results suggest that higher serum IgGI/IgM ratios may be associated with a better clinical response. These changes may reflect dysregulation of immunoglobulin class switching in patients with FSGS. PMID- 9874323 TI - Age and ethnicity affect the risk and outcome of focal segmental glomerulosclerosis. AB - In patients with proteinuria, African-American (AA) ethnicity is reported to be a risk factor for focal segmental glomerulosclereosis (FSGS) and its progression to end-stage renal disease (ESRD). We reviewed our single-center experience to determine the probability of FSGS and its progression to ESRD based on ethnicity and age at presentation in children with proteinuria with or without nephrotic syndrome. Proteinuria without systemic disease or acute glomerulonephritis was the presenting feature in 17% (236/1,403) of children in the renal patient database of Texas Children's Hospital, Baylor College of Medicine. Histopathological diagnoses were established in 107 of 236 patients (45%). FSGS was identified in 65 patients, accounting for 28% of all patients with proteinuria and 61% of patients who underwent renal biopsy. FSGS was more prevalent in AA (45%) than in non-AA patients (22%) (P=0.001), and AA patients with FSGS were older at presentation (12.7+/-4.4 years) than non-AA patients (5.6+/-4.6 years) (P<0.001). Among patients who underwent renal biopsy, increasing age at presentation increased the probability of having FSGS in AA but not non-AA patients (P=0.04). Five-year actuarial renal survival of FSGS was worse in AA (8%) than in non-AA patients (31%) (P=0.01). These data suggest an increased risk and worse outcome of FSGS in AA compared with non-AA children. PMID- 9874324 TI - The role of fludrocortisone in a child with cerebral salt wasting. AB - Cerebral salt wasting (CSW) is a syndrome of hyponatremia due to excessive natriuresis described in patients with central nervous system insult. We present a 29-month-old black male with tuberculous meningitis who developed CSW with depressed mineralocorticoid activity. The patient required hypertonic saline and ionotropic support. Mineralocorticoid supplementation effectively treated CSW. PMID- 9874325 TI - Pleural effusion complicating acute peritoneal dialysis in hemolytic uremic syndrome. AB - Hemolytic uremic syndrome (HUS) is a leading cause of acute renal failure (ARF) in children, and one for which treatment with peritoneal dialysis (PD) is often necessary. Between January 1982 and December 1996, 176 children received PD for ARF at St. Christopher's Hospital for Children; 34 (19%) of whom had HUS. Of these 34, 7 (20%) developed pleural effusions (PE) while receiving PD, whereas none of the remaining 142 children with other causes of ARF did so. The mean age of the 7 affected children was 5.2 (range 0.4-17) years; none had heart failure or nephrotic syndrome, nor had any of them undergone thoracic surgery. PE were diagnosed by chest radiograph at an interval of 2 (range 1-3) days after starting PD. Thereafter, 4 (57%) patients were successfully maintained on a modified PD prescription; 2 others were converted to hemodialysis and 1 to continuous venovenous hemodiafiltration. Although PE are a known complication of PD, none of the patients so treated for non-HUS related ARF developed them. Whether they represent a purely mechanical complication of PD, or are in some way attributable to HUS itself, is not entirely clear. Regardless, when children with HUS require PD, physicians should monitor for the development of this potential complication to minimize the risk of serious respiratory compromise. PMID- 9874326 TI - Non-steroidal anti-inflammatory drug-associated nephrotoxicity in Bartter syndrome. AB - We have followed four patients with Bartter syndrome for a mean of 25.4 years (range 21.5-28.8 years) after diagnosis. All patients received non-steroidal anti inflammatory drugs (NSAID). In all patients, various degrees of renal dysfunction were noted to be temporally associated with NSAID therapy. In two patients, renal dysfunction resolved after discontinuing NSAID therapy, while maintaining other chronic medications such as potassium-sparing diuretics. Renal dysfunction persisted after NSAID withdrawal in two patients. We report these cases as a warning that NSAID should be considered an important cause of either reversible or irreversible renal dysfunction in Bartter syndrome. PMID- 9874327 TI - The use of pamidronate in three children with renal disease. AB - The successful use of pamidronate, a bisphosphonate, for the treatment of hypercalcemia and/or osteopenia is reported in three children with renal failure or following renal transplant. Patient 1 was an 11-year-old post renal transplant male who received a single dose of i.v. pamidronate (0.5 mg/kg) for the treatment of acute hypercalcemia associated with a pathological fracture and subsequent immobilization. Prompt resolution of the hypercalcemia was seen. He received a second course of pamidronate (0.5 mg/kg per day for 3 days) for the treatment of osteopenia and has had a subsequent 15% increase in lumbar spine bone mineral content (BMC). Patient 2, a 14-year-old male on peritoneal dialysis, presented with symptomatic hypercalcemia associated with tertiary hyperparathyroidism. A single dose of i.v. pamidronate (0.4 mg/kg) was given with prompt resolution and prolonged control of his hypercalcemia. The third patient was a 16-year-old female, also in renal failure on peritoneal dialysis. Her course had been complicated by marked osteopenia. I.v. pamidronate (0.5 mg/kg per dose) was given on 3 successive days before and after renal transplant in an attempt to stabilize her bone mineral density (BMD) around the time of renal transplantation, when additional glucocorticoid was necessary. Her total body BMC and BMD remained stable pre and post transplant. The treatment was effective and well tolerated in all three patients. Hence pamidronate is safe and effective for the management of hypercalcemia and osteopenia in children with renal failure and/or renal transplant. PMID- 9874328 TI - Familial, atypical hemolytic-uremic syndrome in a premature infant. AB - The hemolytic-uremic syndrome (HUS) typically presents in toddlers or older children after an episode of bloody diarrhea caused by Escherichia coli O157:H7. However, numerous "atypical" presentations have been described, including familial cases. Here we describe what we believe to be the first report of familial HUS in a premature infant during the neonatal period. PMID- 9874329 TI - Doppler examination of cerebral arteries in uremic children. AB - Transcranial Doppler ultrasonography is a useful method for the estimation and monitoring of cerebral circulation in dialyzed patients. The aim of this study was to evaluate the effect of disease and treatment on cerebral circulation in children on maintenance hemodialysis (HD) and children prior to renal replacement therapy. We demonstrated that in uremic children blood flow velocities of the internal carotid artery (ICA), anterior cerebral artery (ACA), middle cerebral artery (MCA), and posterior cerebral artery (PCA) 120 min and 240 min from the beginning of an HD session were significantly lower than values immediately before HD. Changes in blood flow velocities of MCA and ACA during HD correlated significantly with changes in mean arterial pressure during HD. There was no correlation between changes in blood flow velocities and intradialytic changes in hematocrit values, ultrafiltration, hemoglobin concentration, and blood urea nitrogen values. Mean blood flow velocities of ICA, MCA, and PCA in euvolemic children on conservative treatment were significantly higher than after a HD session in children on maintenance HD. The factors responsible for intradialytic velocity changes of cerebral arteries in uremic children require further examination. PMID- 9874330 TI - Formalin treatment of refractory hemorrhagic cystitis in systemic lupus erythematosus. AB - Hemorrhagic cystitis is a potentially life-threatening complication in systemic lupus erythematosus (SLE). No safe, effective and conservative treatment exists for patients who fail to respond to standard therapy. We report a 17-year-old girl with SLE who suffered from severe hemorrhagic cystitis. Initially, she received frequent red blood cell and platelet transfusions, continuous bladder irrigation, and blood clots were evacuated. Numerous kinds of treatment were tried, including electrocoagulation of bleeding foci, prostaglandin E1 bladder instillation, and hyperbaric oxygen. However, she remained severely anemic and thrombocytopenic necessitating daily transfusions of blood products. After intravesical formalin instillation was performed twice, the hematuria ceased completely. PMID- 9874331 TI - Cell signal transduction through the mitogen-activated protein kinase pathway. PMID- 9874332 TI - Pediatric nephrology workforce in Latin America. AB - Pediatric nephrology workforce issues were examined in a Latin American survey involving 14 countries. The number of children under 15 years per pediatric nephrologist varied widely among countries: Argentina, Cuba, Venezuela, and Uruguay had an unusually high number of pediatric nephrologists. Guatemala represents the opposite end of the spectrum of values (1,582.6 thousand children under 15 years per pediatric nephrologist). A significant inverse correlation was found between children under 15 years per pediatric nephrologist and national gross domestic product per capita (r=-0.52, P<0.05) and a significant correlation between children per pediatric nephrologist and infant mortality (r=0.82, P<0.005, Spearman's rank correlation coefficient). The same correlations were observed for total population per pediatric nephrologist. However, the pediatric nephrology workforce does not merely reflect national economic status. Official health care policies, market forces, and social regulations also have an influence. A study of the number of pediatric nephrologists necessary for adequate planning of care of children with renal disease in Latin America is urgently needed. PMID- 9874333 TI - Changing perspective of lipoid nephrosis in Iranian children. PMID- 9874334 TI - Could this be loin pain hematuria syndrome? PMID- 9874335 TI - Occurrence and survival of verocytotoxin-producing Escherichia coli O157 in raw cow's milk in The Netherlands. AB - From May through November 1997, 1,011 samples of raw milk from bulk storage tanks were examined for the presence of verocytotoxin-producing Escherichia coli of serogroup O157 (O157 VTEC) by immunomagnetic separation following selective enrichment. The samples originated from 1,011 different dairy herds located throughout the Netherlands. O157 VTEC was not isolated from any of the milk samples examined. Additionally, survival of O157 VTEC in raw and UHT-sterilized cow's milk at 7 and 15 degrees C was studied, both in the absence and presence of an activated lactoperoxidase-thiocyanate-hydrogen peroxide system (LPS). Results indicated that the O157 VTEC strain tested was able to grow in raw milk at 7 degrees C as well as at 15 degrees C. Naturally occurring amounts of thiocyanate and hydrogen peroxide in the raw milk tested were not sufficient to activate the LPS. Although the LPS exhibited an antimicrobial activity against O157 VTEC in LPS-activated sterilized milk, O157 VTEC populations were not (or not as obviously) reduced in LPS-activated raw milk. Possibly background microflora were more sensitive to the LPS than the O157 VTEC test strain. It was concluded that raw milk contaminated with O157 VTEC will remain a hazard if kept at 7 or 15 degrees C. Effective pasteurization and avoiding postpasteurization contamination are necessary to ensure the safety of milk. PMID- 9874337 TI - Survival and growth of Campylobacter jejuni after artificial inoculation onto chicken skin as a function of temperature and packaging conditions. AB - Campylobacter jejuni is one of the major causes of food poisoning in humans. C. jejuni is also widespread in food animals, and meat and meat products derived from food animals are the most common vector of bacterial transmission to humans. To determine the role of packing and storage conditions on the replication of C. jejuni on chicken, the virulent strain C. jejuni 81116 was artificially inoculated onto chicken skin pieces (1 cm2) and stored at different temperatures and under various packaging conditions. C. jejuni 81116 remained viable at -20 and -70 degrees C and was able to replicate at 4 degrees C and at ambient room temperature. C. jejuni 81116 was also inoculated onto chicken skin and subjected to repeated freeze thawing and the viability of the inoculum was quantified. C. jejuni 81116 could withstand repeated freeze thawing similar to that which may occur in the domestic home. Under all freezing conditions, C. jejuni 81116 retained a high level of viability and quickly replicated to levels which exceeded Australian food authorities' permitted bacteria level on raw food products after the sample was thawed. PMID- 9874336 TI - Persistence of Escherichia coli O157:H7 in dairy fermentation systems. AB - We examined (i) the persistence of Escherichia coli O157:H7 as a postpasteurization contaminant in fermented dairy products; (ii) the ability of E. coli O157:H7 strains with and without the general stress regulatory protein, RpoS, to compete with commercial starter cultures in fermentation systems; and (iii) the survival of E. coli O157:H7 in the yogurt production process. In commercial products inoculated with 10(3) CFU/ml, E. coli O157:H7 was recovered for up to 12 days in yogurt (pH 4.0), 28 days in sour cream (pH 4.3), and at levels > 10(2) CFU/ml at 35 days in buttermilk (pH 4.1). For the starter culture competition trials, the relative inhibition of E. coli O157:H7 in the experimental fermentation systems was, in decreasing order, thermophilic culture mixture, Lactobacillus delbrueckii subsp. bulgaricus R110 alone, Lactococcus lactis subsp. lactis D280 alone, Lactococcus lactis subsp. cremoris D62 alone, and Streptococcus thermophilus C90 alone showing the least inhibition. Recovery of the rpoS mutant was lower than recovery of its wild-type parent by 72 h or earlier in the presence of individual starter cultures. No E. coli O157:H7 were recovered after the curd formation step in yogurt manufactured with milk inoculated with 10(5) CFU/ml. Our results show that (i) postprocessing entry of E. coli O157:H7 into fermented dairy products represents a potential health hazard; (ii) commercial starter cultures differ in their ability to reduce E. coli O157:H7 CFU numbers in fermentation systems; and (iii) the RpoS protein appears to most effectively contribute to bacterial survival in the presence of conditions that are moderately lethal to the cell. PMID- 9874338 TI - Long-term bacterial profile of refrigerated ground beef made from carcass tissue, experimentally contaminated with pathogens and spoilage bacteria after hot water, alkaline, or organic acid washes. AB - The effects of 2% (vol/vol) lactic acid (LA), 2% (vol/vol) acetic acid (AA), 12% (wt/vol) trisodium phosphate (TSP), 72 degrees C water (HW), and 32 degrees C water (W) washes on bacterial populations which were introduced onto beef carcass surfaces after wash treatments were determined up to 21 days of storage at 4 degrees C of packaged ground beef prepared from the treated and inoculated carcasses. Beef carcass necks were collected from cattle immediately after harvest and subjected to the above treatments or left untreated (control). Neck meat was then inoculated with low levels (ca. <2 log10) of Listeria innocua, Salmonella typhimurium, Escherichia coli O157:H7, and Clostridium sporogenes contained in a bovine fecal cocktail. In general, growth of these four bacteria, aerobic bacteria, lactic acid bacteria, and pseudomonads was suppressed or not observed in the ground beef when LA, AA, or TSP treatments were used as compared to the untreated control. HW or W washes offered little suppression of growth of pathogens during subsequent storage of ground beef when these bacteria were introduced onto beef tissue posttreatment. Of the treatments used, a final LA or AA wash during the processing of beef carcasses offers the best residual efficacy for suppression of pathogen proliferation in ground beef during long-term refrigerated storage or short-term abusive temperature storage if these bacteria contaminate the carcass immediately after carcass processing. PMID- 9874339 TI - Prevalence and genetic variability of Arcobacter species in mechanically separated turkey. AB - A survey for Arcobacter spp. and Arcobacter butzleri in mechanically separated turkey was conducted during the winter of 1995 and summer and fall of 1996. Arcobacter spp. and A. butzleri were identified by polymerase chain reaction and species-specific oligonucleotide probes. Arcobacter spp. were isolated from 77% (303 out of 395) of the mechanically separated turkey samples with 74% (223 out of 303) of these samples positive for A. butzleri. Of the 121 A. butzleri isolates tested, 86 different patterns were evident following amplification of enterobacterial repetitive intergenic consensus sequences. The extent of genetic polymorphism indicated multiple sources of contamination. PMID- 9874340 TI - Survival and growth of psychrotrophic Bacillus cereus in dry and reconstituted infant rice cereal. AB - The potential for growth of enterotoxigenic Bacillus cereus in reconstituted dry foods is a concern, especially when they are consumed by infants or the immunosuppressed. The ability of a four-strain mixture of spores or vegetative cells of psychrotrophic B. cereus to survive in a commercial, dry infant rice cereal as affected by water activity (a(w); 0.27 to 0.28, 0.52 to 0.55, and 0.75 to 0.78), pH (5.6 and 6.7), and temperature (5, 25, 35, and 45 degrees C) was investigated. The rate of death of vegetative cells in dry cereal stored for 36 weeks was not affected by a(w) or pH. Death of spores in cereal stored at 45 degrees C for up to 48 weeks was enhanced at a(w) 0.78 but was unaffected by pH; loss of viability at 5, 25, and 35 degrees C was largely unaffected by differences in a(w). The effect of temperature (8, 15, 21, and 30 degrees C) on outgrowth of spores of B. cereus inoculated at three levels (0.14, 14, and 133 CFU/g, dry weight basis) into cereal reconstituted with apple juice and commercial pasteurized milk (2% fat) was also studied. Outgrowth of spores did not occur in cereal reconstituted with apple juice. Cereal reconstituted with milk and inoculated with 0.14, 14, and 133 spores per g contained >3 log CFU/g within 24, 9, and 6 h, respectively, at 21 degrees C. Populations in cereal reconstituted with milk and inoculated with 133 CFU of B. cereus spores per g reached 7.11, 7.72, and 7.40 log CFU/g within 12, 48, and 72 h when stored at 30, 21, and 15 degrees C, respectively. The organism grew in cereal reconstituted with milk and held at 8 degrees C for 72 h; however, enterotoxin was not detected. In reconstituted cereal inoculated with 133 spores per g, enterotoxin was detected (detection limit 16 ng/g) after 24, 48, and 72 h at 30, 21, and 15 degrees C, respectively, when the population of B. cereus reached >7 log CFU/g. It is recommended that reconstituted infant foods be either consumed immediately or held at < or = 8 degrees C and consumed within 48 h after preparation. PMID- 9874341 TI - Antibiotic susceptibility of potentially probiotic Lactobacillus species. AB - In recent years, the time-honored reputation of lactobacilli as promoters of gastrointestinal and female urogenital health has been qualified. This has occurred due to a rare association with human infection in the presence of certain predisposing factors and their potential to act as a source of undesirable antibiotic resistance determinants to other members of the indigenous microbiota. This necessitates greater caution in their selection for use in microbial adjunct nutrition and disease management (prophylaxis and therapy). It was against this background that 46 Lactobacillus strains from human and dairy sources were assayed for susceptibility to 44 antibiotics. All strains were resistant to a group of 14 antibiotics, which included inhibitors of cell wall synthesis (cefoxitin [30 microg] and aztreonam [30 microg]), protein synthesis (amikacin [30 microg], gentamicin [10 microg], kanamycin [30 microg], and streptomycin [10 microg]), nucleic acid synthesis (norfloxacin [10 microg], nalidixic acid [30 microg], sulphamethoxazole [100 microg], trimethoprim [5 microg], co-trimoxazole [25 microg], and metronidazole [5 microg]), and cytoplasmic membrane function (polymyxin B [300 microg] and colistin sulphate [10 microg]). All strains were susceptible to tetracycline (30 microg), chloramphenicol (30 microg), and rifampicin (5 microg). Four human strains and one dairy strain exhibited atypical resistance to a penicillin, bacitracin (10 microg), and/or nitrofurantoin (300 microg). One human strain was also resistant to erythromycin (15 microg) and clindamycin (2 microg). These resistances may have been acquired due to antibiotic exposure in vivo, but conclusive evidence is lacking in this regard. Seven microorganism-drug combinations were evaluated for beta-lactamase activity using synergy and nitrocefin tests. The absence of activity suggested that cell wall impermeability appeared responsible for beta lactam resistance. The occurrence of a minority of lactobacilli with undesirable, atypical resistance to certain antibiotics demonstrates that not all strains are suitable for use as probiotics or bacteriotherapeutic agents. The natural resistance of lactobacilli to a wide range of clinically important antibiotics may enable the development of antibiotic/probiotic combination therapies for such conditions as diarrhea, female urogenital tract infection, and infective endocarditis. PMID- 9874342 TI - Postprocess contamination of flexible pouches challenged by in situ immersion biotest. AB - Packages were evaluated for leaks by determining microbial penetration through microchannels as a function of test organism concentration, location in a retort, and microchannel diameter and length. A flexible pouch was used in an in situ immersion biotest coupled with a state-of-the-art retort. Microchannel diameters of 10 to 661 microm with 3- and 6-mm lengths were created by placing tungsten wires in vacuum heat-sealed flexible pouches. After removing the wires, these pouches were subsequently heat processed under pressure. They were then biotested in cooling water containing 10(3) and 10(6) CFU of motile Enterobacter aerogenes per ml for 30 min and were dried immediately after manual unloading. After incubation at 37 degrees C for 3 days, they were visually examined for contamination. The high-temperature retorting process was shown to decrease microchannel diameters by an average of 20%. Generally, the smaller the microchannel diameter, the greater the percent shrinkage. Statistical analysis of the biotesting data showed that microchannel diameter and length had strong effects on microbial penetration (P < 0.01). Microbial concentration had a borderline significant effect (P < 0.05), but the effect of package location in the retort was not significant. At conservative conditions, such as a 3-mm microchannel length and a cooling water contamination level of 10(6) CFU/ml, the selected microorganism can penetrate microchannels with diameters as small as 7 microm. However, the minimum microchannel diameter for penetration could be as large as 46 microm at practical conditions of 6-mm microchannel length and contamination levels of 10(3) CFU/ml. PMID- 9874343 TI - Shelf life extension, safety, and quality of fresh pork loin treated with high hydrostatic pressure. AB - The optimal conditions of pressure, time, and processing temperature required to eliminate Listeria monocytogenes Scott A and Salmonella typhimurium ATCC 13311 in fresh pork loin and the effect of these optimal conditions on quality and shelf life were determined. Twenty-five grams of fresh pork loin were inoculated with either of the two organisms and were subjected to pressures between 414 and 827 MPa at either 2 or 25 degrees C for 30 min. The D414MPa(25 degrees C) was determined to be 2.17 min for L. monocytogenes and the D414 MPa (2 degrees C) was determined to be 1.48 min for S. typhimurium. Samples subjected to a 6D process were evaluated by sensory and objective tests as well as for shelf life. These samples were found to be different (P < 0.05) from controls when evaluated after cooking by a triangle test of difference, but only when the pressure was applied at 2 degrees C and not at 25 degrees C. The descriptive analysis test showed that cooked samples treated at 25 degrees C were not different (P > 0.05) from controls in flavor, juiciness, and firmness. Color, peak load, water-holding capacity, and moisture were not found to be different (P > 0.05) between samples treated at 25 degrees C and controls when both were cooked. However, in the raw state, differences were found in the values for color parameters L and b. The level of psychrotrophs was 5.7 log CFU/g for samples treated at 25 degrees C after 33 days of storage at 4 degrees C, as compared with 7.0 log CFU/g for controls. The color and peak load (texture) did not change over the storage period (P > 0.05) in any of the samples. All samples spoiled in 5 days when stored at 25 degrees C. PMID- 9874344 TI - High hydrostatic pressure come-up time and yeast viability. AB - The effects of the come-up time at selected pressures (50 to 689 MPa) on Saccharomyces cerevisiae and Zygosaccharomyces bailii viability were evaluated at 21 degrees C. For Z. bailii the effects of the water activity (a(w)) of the suspension media and the stage of the growth cycle were also investigated. Pressure come-up times exerted an important effect on the yeast survival fraction, decreasing counts as pressure increased. An increased sensitivity to pressure treatments was observed with yeast cells from the exponential growth phase. Lethality increased as a(w) of the suspension media increased. For an a(w) of 0.98 and cells from the stationary growth phase, pressure treatments at less than 200 MPa had no effect on Z. bailii viability; however, no survivors (< 10 CFU/ml) were observed in treatments applied only for the time needed to reach pressures greater than 517 MPa. Yeast survivor curves showed an excellent fit (r > 0.996) when described by a phenomenological model based on the Fermi equation, S(P) = 1/?1 + exp[(P - Pc)/k]?, where S(P) is the survival fraction, P is the pressure, Pc is a critical pressure corresponding to 50% survival, and k is a constant representing the steepness of the curve. PMID- 9874346 TI - The role of trace metal ions in aflatoxin B1 degradation by Flavobacterium aurantiacum. AB - Flavobacterium aurantiacum NRRL B-184 possesses the ability to degrade aflatoxin B1 in solution and in several food items. Aflatoxin B1 is a potent carcinogen that causes significant economic losses to the agricultural and food industry. The role of trace metal ions (Cu2+, Mn2+, Zn2+, and Co2+) were studied in an effort to understand the enzymatic system involved in aflatoxin B1 degradation by F aurantiacum. The effect of divalent chelators (EDTA and 1,10-phenanthroline [OPT]) in the presence of the trace metal ions was studied as well. Aflatoxin B1 (10 microg/ml) was added to 72-h cultures of F aurantiacum that had been washed and resuspended in phosphate buffer (pH 7.0). HPLC was used to determine aflatoxin B1 concentration in these cultures. Incubating cells at 30 degrees C with 1 and 10 mM Cu2+, Mn2+, and Zn2+ significantly decreased aflatoxin B degradation after 4 and 24 h (P < 0.05). Decreased degradation was also observed with 1 and 10 mM Cu2+ and Zn2+ after 48 h and with 0.1 mM Cu2+ after 24 and 48 h. Co2+ did not have a significant effect on aflatoxin B1 degradation. EDTA and OPT did not counter the inhibition in the presence of Cu2+. The addition of 1 mM EDTA countered the inhibition by 1 mM Mn2+ after 4 and 24 h, but 1 mM OPT did not counter the inhibition by 10 mM Mn2+ after 4 and 24 h. OPT countered the inhibition by 1 mM Zn2+ after 4 and 48 h. These trace elements inhibit aflatoxin B1 degradation by F aurantiacum. In addition, their presence necessitates higher concentrations (>1 mM) of EDTA and OPT for the removal of their inhibitory effect. PMID- 9874345 TI - Biological control of Botrytis cinerea growth on apples stored under modified atmospheres. AB - The combined effect of modified-atmosphere packaging and the application of a bacterial antagonist (Erwinia sp.) on Botrytis cinerea growth on apples (cv. 'Golden Delicious') was investigated. Inoculated apples were stored in polyethylene bags at 5 degrees C. The initial gas composition in each bag was set according to a central composite experimental design involving five levels of O2 (1 to 15%) and CO2 (0 to 15%). Control samples under ambient conditions were also included. Without the antagonist, measurements of mold colony diameter over time showed that O2 had no effect on the growth of B. cinerea, while increased CO2 levels delayed its growth by about 4 days. Application of the antagonist resulted in a significant interaction between O2 and CO2. At low O2 levels, CO2 had no effect on mold growth, but at high O2, CO2 enhanced mold growth. O2 and the antagonist worked synergistically to reduce mold growth by about 6 days at low levels of CO2. However, at high CO2 levels, O2 had no effect. The strongest antagonistic effect was observed under ambient conditions. Overall, results showed that high CO2 atmospheres can slow the growth of B. cinerea and that Erwinia sp. was an effective antagonist against B. cinerea growth on apples, particularly under ambient conditions. PMID- 9874347 TI - Occurrence of aflatoxins, fumonisin B1, and zearalenone in foods and feeds in Botswana. AB - Sorghum and maize form the main dietary staple foods in Botswana. Other products such as peanuts, peanut butter, phane (an edible larval stage of an emperor moth Imbrasia belina Westwood), and pulses (cowpeas and beans) are also widely used as food and for the manufacture of feeds. These important food and feed commodities were analyzed for the presence of aflatoxins, fumonisin B1, and zearalenone. Aflatoxins were detected in 40% of the samples analyzed. The concentration of total aflatoxins ranged from 0.1 to 64 microg/kg. The mean concentration ranged from 0.3 microg/kg in sorghum to 23 microg/kg in peanut butter. Peanut butter samples were the most contaminated (71%). No aflatoxins were detected in maize. Fumonisin B1 was detected in 36% of the samples. Maize samples were the most contaminated (85% of the samples) with the concentration ranging from 20 to 1,270 microg/kg. No fumonisin B1 was detected in peanuts, phane, and beans. Zearalenone was only found in 2.6% of the samples analyzed at 40 microg/kg. Aflatoxins were the most common toxins detected in foods and feeds in Botswana. However, fumonisin B1 was more prevalent in maize than aflatoxins or zearalenone. PMID- 9874349 TI - Thiamin, riboflavin, and alpha-tocopherol content of exotic meats and loss due to gamma radiation. AB - Changes in thiamin, riboflavin, and alpha-tocopherol concentrations due to gamma irradiation were followed in alligator, caiman, bison, and ostrich (exotic) meats. The proximate composition showed that the exotic meats generally had lower fat content than domestic animal meats and that the thiamin content of the reptiles was lower. The changes in the vitamins due to irradiation were similar to those previously observed for domestic species. The results indicate that the loss of vitamins in these species is negligible insofar as the American diet is concerned, and that the concept of "chemiclearance" is applicable to exotic meats. PMID- 9874348 TI - Distribution of Norwalk virus within shellfish following bioaccumulation and subsequent depuration by detection using RT-PCR. AB - Consumption of raw bivalve mollusks contaminated with pathogens from human feces continues to present a human health risk. The purpose of this study was to monitor the uptake, localization, and removal of Norwalk virus (NV) in shellfish (oyster and clam) tissues by analyzing virus distribution in selected dissected tissues. Live shellfish were allowed to bioaccumulate different input titers of NV for time periods from 4 to 24 h. In some experiments, depuration by shellfish that bioaccumulated NV and Escherichia coli bacteria was allowed to proceed for 24 or 48 hours. Dissected stomach (St), digestive diverticula (DD), adductor muscle (AM), and hemolymph cells (HC) tissues were assayed for NV by the reverse transcription polymerase chain reaction (RT-PCR) method. An internal RNA standard control was added to the RT-PCR to identify the presence of inhibitors to RT-PCR. NV titers in DD tissues before and after depuration were estimated using quantitative RT-PCR end-point dilution. NV was found in the alimentary tract (DD or St) at all concentrations of input virus, but was present more frequently after exposure to higher levels of virus. NV was detected in AM and HC only following exposure to higher levels of virus. In experiments where depuration by oysters was continued for 48 h, depuration of bacteria was efficient (95% reduction of bacteria), but minimal (7%) reduction of NV titers from DD tissues was detected. These findings indicate that NV can localize both within and outside the alimentary tract of shellfish, and NV is poorly depurated using conditions favorable for E. coli depuration. PMID- 9874350 TI - Polymerase chain reaction-restriction fragment length polymorphism analysis of a short fragment of the cytochrome b gene for identification of flatfish species. AB - Restriction site analysis of polymerase chain reaction (PCR) products from a conserved region of the cytochrome b gene has been used for the specific identification of sole (Solea solea), European plaice (Pleuronectes platessa), flounder (Platichthys flesus), and Greenland halibut (Reinhardtius hippoglossoides). PCR amplification of the cytochrome b gene using a universal primer together with a primer specifically designed as a part of this study produced a 201-bp fragment in all species analyzed. Digestions of the PCR products with Sau3Al, BsmAl, Rsal, and Mn/l endonucleases, followed by agarose gel electrophoresis of the digested PCR products, yielded specific profiles that enabled direct identification of each species analyzed. PMID- 9874351 TI - Quantitative evaluation of pork adulteration in raw ground beef by radial immunodiffusion and enzyme-linked immunosorbent assay. AB - Quantitative estimates are important to establish whether pork adulteration in ground beef is accidental or intentional. A standard agar gel radial immunodiffusion (RID) test using forensic-grade antiserum to porcine albumin and an enzyme-linked immunosorbent assay (ELISA) using forensic-grade anti-porcine glycoprotein immunoglobulin were used to determine from 1 to 75% raw pork in raw ground beef. The RID test, which incorporated 1.5% anti-pork serum in 1% immunodiffusion agar, formed precipitin rings with pork albumin in agar wells. A linear standard curve was obtained by plotting the diffusion area against standard pork concentrations ranging from 0 to 80%. For the ELISA the endpoint optical density increased linearly versus log % pork between 0.0625% and 2% pork. In spiked samples, the RID test had a detection limit of 3 to 5%, a coefficient of variation (CV) of 22%, and a recovery of 105%. The ELISA had a detection limit of 1%, a CV of 18%, and a recovery of 114%. The mean recovery from the spiked samples by the ELISA and RID test was not significantly different (P > 0.05) from the known sample amounts. Quantitation by RID of 28 ground beef samples (27 of which were DTEK ELISA-positive for pork adulteration) revealed a wide range of pork content, with values as high as 48%. PMID- 9874352 TI - Detection of bovine milk proteins in soymilk by Western blotting. AB - A Western blotting method for the detection of whey milk proteins in commercial soymilks was applied to assess the food safety. Soy proteins and milk proteins were separated by SDS-PAGE in PhastSystem equipment. After the electrophoretic separation, immunodetection with anti-bovine alpha-lactalbumin and anti-bovine beta-lactoglobulin antisera was performed. Adulteration with bovine protein in percentages of 0.1% in soy protein can be detected. Western blotting of bovine alpha-lactalbumin and bovine beta-lactoglobulin was applied to detect adulteration by bovine milk proteins in different soymilks: powdered soymilk and soy infant formulas. PMID- 9874353 TI - Comparison of gas chromatography-electron capture and enzyme-linked immunosorbent assay for deoxynivalenol in milled fractions of naturally contaminated wheat. AB - An enzyme-linked immunosorbent assay (ELISA) for deoxynivalenol was compared with a gas chromatography-electron capture assay to determine deoxynivalenol levels in milled fractions of wheat. The milling provided eight fractions: first, second, and third break flours; first, second, and third reduction flours; brans; and shorts. The difference between levels of deoxynivalenol quantitated by ELISA or gas chromatography did not depend significantly on the wheat samples or milled wheat fractions. For none of the fractions or samples did the differences differ significantly (P=0.05) from zero. Based on these comparative tests, ELISAs for deoxynivalenol in milled wheat fractions should provide reliable results rapidly and economically in a commercial setting. PMID- 9874354 TI - Filterability of autotransfusion blood cells and plasma after total knee arthroplasty. AB - Postoperatively filtered drain blood can be used for autotransfusion, but quality has been questioned. The blood contains activated or destroyed blood cells, debris, and chemicals from the wound. The ConstaVac autotransfusion system was used in 18 patients operated with a total knee prosthesis. Samples were taken from drain blood and compared with venous blood at different sampling times (to 24 h) and different incubation conditions. Micro-rheology was measured as pore filterability (5 microm) in whole blood, erythrocyte resuspension, and plasma. The filtration flow resistance was significantly lower in drain whole blood, but was unchanged in erythrocyte resuspension compared to venous blood. However, the rate of filter clogging was unchanged in drain whole blood, but was increased in the erythrocyte resuspension at 2 h. Drain plasma filterability indicated particle contamination. In total, the filterability of resuspended drain erythrocytes and cell-free plasma is affected, but this is not significant enough to have effects on drain whole blood filterability which is increased, possibly due to the leukocyte reduction. From these perspectives, we consider drain blood acceptable for autotransfusion. PMID- 9874355 TI - Opposite effects of low-density and high-density lipoprotein on blood viscosity in fasting subjects. AB - Given the enlarging body of evidence implicating increased blood viscosity in atherogenesis, the authors hypothesize that lipoproteins modulate the atherogenic process by affecting blood viscosity. In order to define the magnitude of the effect of lipoproteins on blood viscosity, capillary viscometry was performed on blood from 16 healthy, fasting subjects, and results were correlated with lipoprotein-cholesterol levels. Low-density lipoprotein-cholesterol was positively associated with blood viscosity (r = 0.610, p = 0.01). High-density lipoprotein-cholesterol was negatively associated with blood viscosity (r = 0.479, p = 0.06). A multiple regression model was developed with these data, revealing that 54% of variation in blood viscosity was attributable to these lipoproteins. This model was validated on a second dataset, in which these lipoproteins accounted for 28% of variation in blood viscosity. A second model, including hematocrit, serum viscosity, and high-density lipoprotein-cholesterol levels, explained 73% of variation in blood viscosity. By modulating blood viscosity and flow, lipoproteins may affect the residence time of atherogenic particles and atherogenesis. PMID- 9874356 TI - Loading of intact rabbit erythrocytes with fluorophores and the enzyme pronase by means of electroporation. AB - The application of electric field pulses in cell suspensions is known to alter membrane integrity, resulting in increased membrane permeabilization. This field induced membrane poration provides the means to load cells with a variety of external substances, useful for clinical applications. In this work, intact rabbit erythrocytes were successfully loaded with low molecular weight fluorescent probes and with the high molecular weight enzyme pronase, which has been shown to mimic the effects of insulin. Attachment of the enzyme onto the cell surface was also achieved by modifying the applied pulse parameters. Both applications were efficient and accompanied by high cell survival rates. In this way, biological carriers loaded with active substances were produced, offering the potentials for useful clinical applications, either for diagnostic or therapeutic purposes. PMID- 9874357 TI - Fibrinogen is negatively correlated with aerobic working capacity in football players. AB - While it is well established that blood viscosity is decreased in sportsmen and related to fitness, the involvement of fibrinogen in this relationship is less well defined. Relationships among fitness, rheology and fibrinogen were investigated in 32 football players (age 17-33 years: 19 professionals and 13 leisure players). A submaximal 25 min exercise-test was performed and allowed the calculation of aerobic working capacity. Aerobic working capacity (W170 and VO2 max) was negatively correlated to fibrinogen (r = -0.531, p < 0.01 and r = 0.623, p < 0.01), while on the whole sample the correlation to viscosity and erythrocyte aggregation was not significant. When subjects were divided into two subgroups according to their plasma fibrinogen concentration, the aerobic working capacity (either expressed as W170 or VO2 max) is higher when plasma fibrinogen level is lower than 2.7 g/l. Thus, there is a highly significant negative correlation between fibrinogen and fitness in these sportsmen, independent of blood rheology. These data suggest that rheology and fibrinogen are to some extent separate determinants of an individual's fitness. PMID- 9874358 TI - Rectification on the inertial effects for co-axial cylinders rheometer. AB - The inertia of the fluid and the inner cylinder in co-axial cylinders rheometer have a great influence on the unsteady flow of non-Newtonian fluid. In the paper this phenomenon is studied minitely and a method to correct the measured results for an unsteady flow in the rheometer is proposed. It was found that the inertia effect of a fluid can be ignored when the gap between cylinders is smaller and the inertia effect of the inner cylinder can be represented with a vibration equation. Using the equation changes of Huang's blood constitutive parameters are observed. PMID- 9874359 TI - A new model to assess tcpO2 changes during heat-dependent vasodilation: preliminary observations. AB - The transcutaneous pO2 (tcpO2) changes induced by the increase of temperature to 45 degrees C after electrode stabilization at 37 degrees C were continuously recorded. The mathematical analysis of the curve led us to identify three different parameters: the half-slope "b" time, the angular "mT" coefficient and the angular "mb" parameter, which are not simply the expression of the maximum vasodilation capacity but mainly of the dynamic response of the microcirculatory system to skin heating. These parameters derived from the computerized mathematical analysis of the curve can be also obtained by a graphical approach with a very good approximation. The angular "mb" coefficient was shown to possess the highest statistical power in discriminating the POAD patients from normal controls. The clinical relevance and the possible pathophysiological meaning of these preliminary observations have to be confirmed. PMID- 9874360 TI - Relationships among body composition, hemorheology and exercise performance in rugbymen. AB - We investigated relationships among body composition, blood rheology, and exercise performance in 14 rugbymen (19-31 yr, weight 65.8-109.2 kg, height 1.7 1.96 m, body mass index 21.7-33.1 kg/m2) who underwent a standardized submaximal exercise session on cycloergometer corresponding to 225 kJ over 30 min. The rheologic response to exercise was measured with the MT90 viscometer and the Myrenne aggregometer. Dehydration, evaluated by precision weighing, resulted in a loss of 360 to 973 g water, i.e., 1.69 to 4.32 g/kJ. This loss of water is not correlated to plasma volume contraction as assessed by the equation of Greenleaf. Hemorheologic changes are observed, but they are correlated neither to water loss, nor to plasma volume contraction. A 36% increase in blood viscosity (p < 0.01) is mainly explained by a red blood cell rigidification (p < 0.02), although hematocrit and plasma viscosity also increase (p < 0.01). Isometric adductor strength (specific ergometer) is correlated to erythrocyte flexibility (r = 0.680, p < 0.01). Red cell aggregability (Myrenne aggregometer) is correlated to fat mass measured by bioelectrical impedance (r = 0.634, p < 0.02). Aerobic working capacity index W170 is negatively correlated to the increase in plasma viscosity during exercise (r = -0.546, p < 0.05), suggesting that this event is less important in stronger individuals. This study shows that fat mass, even within a physiological range, is a determinant of erythrocyte aggregability, suggesting that training-induced alterations in body composition play a role in the specific hemorheologic profile of athletes. In addition, both erythrocyte flexibility and the magnitude of fluid shifts during exercise appear to be related to fitness in these sportsmen. PMID- 9874361 TI - Erythrocyte elongation index measured on a Rheodyn SSD laser diffractometer. Influence of the hematocrit. PMID- 9874363 TI - Europe bids for molecular biology 'club'. PMID- 9874362 TI - Hemorheological changes in children with polygenic hypercholesterolemia. AB - In order to ascertain whether polygenic hypercholesterolemia (PH), the most common cause of small increases in plasma lipids during childhood, is associated with rheological alterations, we determined the hemorheological and lipid profile of 21 PH children (12 males, 9 females) and a well-matched control group (CG). In addition, a carotid ultrasound was done on all the PH children, but showed no alterations. When compared with the CG, PH children showed increased erythrocyte aggregation both at stasis (EAM0) (4.39+/-1.15 vs. 3.75+/-1.02), p < 0.05, and at low shear rate (EAM1) (8.22+/-1.42 vs. 7+/-1.39), p < 0.01, and increased plasma viscosity (PV) (1.19+/-0.04 cP vs. 1.15+/-0.04 cP), p < 0.01. These results reinforce the hypothesis that lipid metabolic alterations are associated with specific rheological modifications in absence of a demonstrable atherosclerotic lesion. PMID- 9874364 TI - South Korean researchers under fire for claims of human cloning. PMID- 9874365 TI - Canadian whistleblower row prompts broader code of conduct. PMID- 9874366 TI - Longevity and the barren aristocrat. PMID- 9874367 TI - Auditory perception. Sounds in a virtual world. PMID- 9874368 TI - Anoxic bioremediation of hydrocarbons. PMID- 9874369 TI - Human longevity at the cost of reproductive success. AB - The disposable soma theory on the evolution of ageing states that longevity requires investments in somatic maintenance that reduce the resources available for reproduction. Experiments in Drosophila melanogaster indicate that trade-offs of this kind exist in non-human species. We have determined the interrelationship between longevity and reproductive success in Homo sapiens using a historical data set from the British aristocracy. The number of progeny was small when women died at an early age, increased with the age of death, reaching a plateau through the sixth, seventh and eighth decades of life, but decreased again in women who died at an age of 80 years or over. Age at first childbirth was lowest in women who died early and highest for women who died at the oldest ages. When account was taken only of women who had reached menopause, who were aged 60 years and over, female longevity was negatively correlated with number of progeny and positively correlated with age at first childbirth. The findings show that human life histories involve a trade-off between longevity and reproduction. PMID- 9874370 TI - Role of spectral detail in sound-source localization. AB - Sounds heard over headphones are typically perceived inside the head (internalized), unlike real sound sources which are perceived outside the head (externalized). If the acoustical waveforms from a real sound source are reproduced precisely using headphones, auditory images are appropriately externalized and localized. The filtering (relative boosting, attenuation and delaying of component frequencies) of a sound by the head and outer ear provides information about the location of a sound source by means of the differences in the frequency spectra between the ears as well as the overall spectral shape. This location-dependent filtering is explicitly described by the head-related transfer function (HRTF) from sound source to ear canal. Here we present sounds to subjects through open-canal tube-phones and investigate how accurately the HRTFs must be reproduced to achieve true three-dimensional perception of auditory signals in anechoic space. Listeners attempted to discriminate between 'real' sounds presented from a loudspeaker and 'virtual' sounds presented over tube phones. Our results show that the HRTFs can be smoothed significantly in frequency without affecting the perceived location of a sound. Listeners cannot distinguish real from virtual sources until the HRTF has lost most of its detailed variation in frequency, at which time the perceived elevation of the image is the reported cue. PMID- 9874371 TI - Hedgehog stimulates maturation of Cubitus interruptus into a labile transcriptional activator. AB - In Drosophila, signalling by the protein Hedgehog (Hh) alters the activity of the transcription factor Cubitus interruptus (Ci) by inhibiting the proteolysis of full-length Ci (Ci-155) to its shortened Ci-75 form. Ci-75 is found largely in the nucleus and is thought to be a transcriptional repressor, whereas there is evidence to indicate that Ci-155 may be a transcriptional activator. However, Ci 155 is detected only in the cytoplasm, where it is associated with the protein kinase Fused (Fu), with Suppressor of Fused (Su(fu)), and with the microtubule binding protein Costal-2. It is not clear how Ci-155 might become a nuclear activator. We show here that mutations in Su(fu) cause an increase in the expression of Hh-target genes in a dose-dependent manner while simultaneously reducing Ci-155 concentration by some mechanism other than proteolysis to Ci-75. Conversely, eliminating Fu kinase activity reduces Hh-target gene expression while increasing Ci-155 concentration. We propose that Fu kinase activity is required for Hh to stimulate the maturation of Ci-155 into a short-lived nuclear transcriptional activator and that Su(fu) opposes this maturation step through a stoichiometric interaction with Ci-155. PMID- 9874372 TI - Local calcium signalling by inositol-1,4,5-trisphosphate in Purkinje cell dendrites. AB - The second messenger inositol-1,4,5-trisphosphate (InsP3) releases Ca2+ from intracellular Ca2+ stores by activating specific receptors on the membranes of these stores. In many cells, InsP3 is a global signalling molecule that liberates Ca2+ throughout the cytoplasm. However, in neurons the situation might be different, because synaptic activity may produce InsP3 at discrete locations. Here we characterize InsP3 signalling in postsynaptic cerebellar Purkinje neurons, which have a high level of InsP3 receptors. We find that repetitive activation of the synapse between parallel fibres and Purkinje cells causes InsP3 mediated Ca2+ release in the Purkinje cells. This Ca2+ release is restricted to individual postsynaptic spines, where both metabotropic glutamate receptors and InsP3 receptors are located, or to multiple spines and adjacent dendritic shafts. Focal photolysis of caged InsP3 in Purkinje cell dendrites also produces Ca2+ signals that spread only a few micrometres from the site of InsP3 production. Uncaged InsP3 produces a long-lasting depression of parallel-fibre synaptic transmission that is limited to synapses where the Ca2+ concentration is raised. Thus, in Purkinje cells InP3 acts within a restricted spatial range that allows it to regulate the function of local groups of parallel-fibre synapses. PMID- 9874373 TI - A new class of synaptic response involving calcium release in dendritic spines. AB - In the classical view, transmission of signals across synapses in the mammalian brain involves changes in the membrane potential of the postsynaptic cell. The use of high-resolution cellular imaging has revealed excitatory synapses at which postsynaptic, transient alterations in calcium ion concentration are tightly associated with electrical responses. Here, by investigating the synapse between parallel glutamatergic fibres and Purkinje cells in the mouse cerebellum, we identify a class of postsynaptic responses that consist of transient increases in dendritic Ca2+ concentration but not changes in somatic membrane potential. Our results indicate that these synaptic Ca2+ transients are mediated by activation of metabotropic glutamate-responsive mGluR1-type receptors and require inositol 1,4,5-trisphosphate-mediated Ca2+ release from intradendritic stores. The new type of synaptic response is restricted to postsynaptic microdomains, which range, depending on the frequency of stimulation, from individual spines to small spinodendritic compartments. Thus, the synaptic Ca2+-release signal may be one of the critical cues that determine the input specificity of long-term depression, a well-established form of activity-dependent plasticity at these synapses. PMID- 9874374 TI - Creating statewide tobacco control programs after passage of a tobacco tax: executive summary. AB - The tobacco control leaders of four states that have successfully passed tobacco tax ballot initiatives (California, Massachusetts, Arizona, and Oregon) were brought together to this conference to discuss what happens after the passage of such an initiative, how to defend tobacco control funds against diversion for other purposes by a state's legislature, and how to use the revenues maximally to reduce the health and economic toll taken by tobacco use. At the conference, the attendees from each state presented reports that will undoubtedly be useful to other states seeking to pass their own tobacco tax legislation or initiatives. PMID- 9874375 TI - Tobacco tax initiatives to prevent tobacco use: a study of eight statewide campaigns. AB - This article reviews the history of successful and unsuccessful tobacco tax initiatives in eight states in the U. S. since 1988. It addresses the common origins of these initiatives and proposes several strategies for the success of citizen-based initiative campaigns attempting to raise the tobacco excise tax. It explores the impact of tobacco tax increases on youth and discusses why youth consumption is increasing even in the face of rising tobacco taxes. Only 50% of the states in the U. S. can pass tobacco tax increases using the initiative process; the other states require legislative action. Four states have succeeded in passing citizen-sponsored tobacco tax initiatives, whereas two others have failed at the ballot. Efforts in two other states foundered when insufficient signatures were submitted to gain a spot on the ballot. Surveys in all six states in which initiatives were placed on the ballot revealed similar high levels of voter support, but the clearest factor separating winning from losing campaigns was the availability of sufficient financial resources. Other important campaign elements included strong leadership, broad coalitions, experienced legal and political consultants, access to public opinion research, and advance planning. PMID- 9874376 TI - History of successful ballot initiatives--California. AB - In this report, the author examines the factors that led to a tobacco tax ballot initiative in California, the coalition-building process, the strong arguments for tax increases, and important decisions regarding the distribution of revenues. PMID- 9874377 TI - History of successful ballot initiatives--Massachusetts. AB - In this report, the author traces the Massachusetts campaign for the 1992 referendum Question 1, which raised the cigarette excise tax by 25 cents. Dr. Cady highlights the major features of the 2-year campaign with special emphasis on the need for committed leadership, coalition-building, and expert political guidance. PMID- 9874378 TI - History of successful ballot initiatives--Arizona. AB - In this article, the author, a key architect of the successful tobacco control program in Arizona, examines the stages of preparing an initiative campaign and the major issues in recruiting hospital associations to antitobacco campaigns. The article provides important lessons for coalitions and hospital associations, potential key players in other statewide campaigns. PMID- 9874379 TI - History of successful ballot initiatives--Oregon. AB - In this report, the author traces the background of Ballot Measure 44, which raised taxes on cigarette and other tobacco products in Oregon. He highlights the effect of a positive coalescing of different forces (nonprofit health groups and medical groups) despite initial differences in program strategy. PMID- 9874380 TI - Getting key players to work together and defending against diversion--California. AB - In 1988, Californians passed Proposition 99, which raised the tobacco tax from 10 cents to 35 cents and allocated 20% of the resulting revenues, approximately $100 million, for tobacco education and 5% for tobacco-related research. Seventy percent was allocated for health care costs and 5% for the environment. In this report, the authors provide an overview of the process of implementing the tobacco education program, which is followed by a more detailed review and analysis of the major impediments to the program, i.e., the diversion of a significant portion of the funding voters had earmarked for tobacco education and research and the battles for legislative reauthorization of Proposition 99 in the 1990s. PMID- 9874381 TI - Getting key players to work together and defending against diversion- Massachusetts. AB - In this report, the authors trace the development of an effective statewide tobacco control program following victory at the Massachusetts polls in 1992. They examine the pitfalls standing in the way of coalition building and present important lessons for states in similar positions. PMID- 9874382 TI - Getting key players to work together and defending against diversion--Arizona. AB - In this report, the author traces the development of the offensive and defensive strategies employed by various antitobacco coalitions and organizations in Arizona. Further, he describes key principles and identifies directions for states soon to embark on tobacco control initiatives. PMID- 9874383 TI - Getting key players to work together and defending against diversion--Oregon. AB - In this report, the author, Executive Vice President of the American Cancer Society in Oregon, reviews the key factors in the crafting of the coalition leading to passage of a tobacco tax in Oregon. Mr. North also emphasizes the importance of institutionalizing the coalition after passage to provide the required leadership for future efforts. PMID- 9874384 TI - Designing an effective statewide tobacco control program--California. AB - In this report, the author traces the history of the development of the first statewide tobacco control program, from its early obstacles and legislative imposed deadline through the development of its three priority areas. Woven into this review is Dr. Bal's emphasis on the role of a cohesive leadership tied into the needs of the tobacco control constituency. PMID- 9874385 TI - Designing an effective statewide tobacco control program--Massachusetts. AB - Smoking-related illnesses kill > 10,000 Massachusetts residents each year and cost hundreds of millions of dollars of public and private expenditures for health care. To combat this public health problem, in 1992 Massachusetts voters approved a referendum question calling for an increased excise tax on tobacco products, with the revenue supporting a Health Protection Fund. Approximately 40% of the fund is used to finance the Massachusetts Tobacco Control Program (MTCP), administered by the Massachusetts Department of Public Health. During the first 3 fiscal years (FY), the MTCP budget has averaged just over $40 million annually, declining during that period from approximately $43 million in FY 1995 to < $37 million in FY 1997. PMID- 9874386 TI - Designing an effective statewide tobacco control program--Arizona. AB - In this article, the author examines the creation and development of Arizona's statewide tobacco control program. In particular, he examines the three basic goals of prevention, cessation, and policy and emphasizes the need to focus equally on all to build an effective program. The need for sound theory and research to guide statewide comprehensive programs is highlighted. PMID- 9874387 TI - Designing an effective statewide tobacco control program--Oregon. AB - In this report, the author documents the first stages in the development and implementation of Oregon's tobacco control program. She briefly reviews the history of tobacco control coalitions in Oregon prior to the passage of Ballot Measure 44 and attributes much of the success to earlier coalition-building efforts. PMID- 9874388 TI - Designing an effective counteradvertising campaign--California. AB - In this report, the author shares some of the basic lessons learned in conducting a tobacco education and prevention media campaign for the state of California over the past 9 years. She highlights the three most fundamental lessons: 1) the qualities to look for in an advertising/public relations agency, 2) the importance of integrating a media campaign with other tobacco control activities, and 3) the most successful advertising strategies. PMID- 9874389 TI - Designing an effective counteradvertising campaign--Massachusetts. AB - In this report, the author reviews the major lessons from the Massachusetts tobacco counteradvertising efforts. Ms. Miller ties in the Massachusetts three pronged campaign of youth prevention, adult cessation, and public support with focus group-tested counteradvertising strategies. PMID- 9874390 TI - Designing an effective counteradvertising campaign--Arizona. AB - In this report, the authors describe an innovative antitobacco advertising campaign in Arizona that was aimed at youth. The campaign utilizes traditional as well as non-traditional methods of research to determine the attitudes, behavior and values of today's adolescents. These creative methodologies led to rigorously tested and well-received media strategies. PMID- 9874391 TI - Designing an effective counteradvertising campaign--Oregon. AB - In this report, the authors describe the key components for development of the soon to be created counteradvertising campaign. Emphasis is placed on numerous factors, including training, selection of target groups, the need for multiple media channels, and scientific assessment of media strategies. PMID- 9874392 TI - Evaluation: methods and strategy for evaluation--California. AB - In this report, the author documents the many successes in California's history of tobacco control efforts, and the primary role given to evaluation. She provides a comprehensive overview of California's multidimensional evaluation structure and concludes with key lessons gleaned from a decade of evaluation research. PMID- 9874393 TI - Evaluation: methods and strategy for evaluation--Massachusetts. AB - Various challenges exist when evaluating statewide, community-based public health initiatives and these need to be addressed by tobacco control programs. There are factors inherent in public health initiatives that can make it difficult to assess causality and effectiveness. These include initial design elements, funding mandates, duration of program and resources allocated for evaluation. In addition, outside efforts to undermine the initiative also must be taken into consideration, when evaluating effectiveness. This article focuses on these and other challenges encountered by the Massachusetts Tobacco Control Program and presents information on the state specific tobacco surveys conducted for the Program. PMID- 9874394 TI - Evaluation: methods and strategy for evaluation--Arizona. AB - In this report, the author describes a relatively new but novel approach for program evaluation currently underway in Arizona. The author notes the crucial importance of local initiatives in tobacco control buoyed by evaluations integrally connected with day-to-day programs and led by local staff. PMID- 9874395 TI - Evaluation: methods and strategy for evaluation--Oregon. AB - The authors describe the development of comprehensive mechanisms for the evaluation of Oregon's newly funded tobacco control efforts through building on established surveillance systems. PMID- 9874396 TI - Key tobacco tax references and resources. PMID- 9874397 TI - Incidence of breast carcinoma-related lymphedema. AB - BACKGROUND: Of the 2 million breast carcinoma survivors, perhaps 15-20% are living currently with posttreatment lymphedema. Along with the physical discomfort and disfigurement, patients with lymphedema also must cope with the distress derived from these symptoms. METHODS: To review the medical literature for the question of lymphedema incidence, a comprehensive, computerized search was performed. All publications with subject headings designating breast carcinoma-related lymphedema from 1970 to the present (116 reports) were found, and each summary or abstract was read. Of the 116 reports, 35 discussed the incidence of lymphedema. Of these, seven reports since 1990 from five countries with the most relevance to current patients were then chosen for greater analysis and comparison. RESULTS: The incidence of lymphedema ranged from 6% to 30%. The source of patients, length of follow-up, measurement techniques, and definition of lymphedema varied from report to report. In general, reports with shorter follow-up reported lower incidences of lymphedema. CONCLUSIONS: The definitive study to determine the incidence of lymphedema has not been performed to date. There has been no prospective study in which patients have been followed at intervals with accurate measurement techniques over the long term. PMID- 9874398 TI - Surgical treatment and lymphedema. AB - BACKGROUND: Lymphedema is a serious and disabling complication of the treatment of breast carcinoma. This is related directly to the removal of axillary lymph nodes. Because lymph node status is the single most important predictor of outcome, it is necessary to obtain accurate information. Whereas breast conservation has become the preferred approach for treating early breast carcinoma, the accompanying axillary dissection continues to cause morbidity. METHODS: The history of the operations for breast carcinoma is reviewed, and the anatomy, techniques, and complications of axillary lymphadenectomy are described. Data to support the necessity for accurate axillary staging is presented, and results of noninvasive axillary staging approaches are discussed. The technique and value of sentinel node biopsy are presented. RESULTS: Axillary lymphadenectomy is required where lymph node metastases are present to accomplish local control, improve survival, and provide information for staging to plan adjunctive therapy. Noninvasive techniques do not yet provide high enough sensitivity to assess the status of the axilla. The sentinel lymph node biopsy is a technique that can identify those patients who require axillary lymphadenectomy. CONCLUSIONS: Screening mammography has been responsible for down staging the size of detected breast carcinomas, so that the axillary dissection may be omitted in small carcinomas of favorable histologic type. For carcinomas in which the probability of axillary metastases exists, by using the sentinel lymph node biopsy, axillary dissections can be avoided when results are negative, and the risk of lymphedema can be reduced. PMID- 9874399 TI - Breast radiotherapy and lymphedema. AB - BACKGROUND: Breast radiotherapy has a low incidence of long term complications. Lymphedema is the most commonly reported complication and adversely affects the quality of life of the breast carcinoma patient. Although its incidence is decreasing, lymphedema still remains a significant concern for patients and their physicians. With the indications for radiotherapy in breast carcinoma management broadening, current strategies to prevent radiation-related lymphedema should be applied and new strategies should be developed. METHODS: A review of the literature addressing lymphedema as a complication of radiotherapy in breast carcinoma management was performed. RESULTS: Arm, breast, and truncal edema occur after primary breast carcinoma management. The literature supports the view that radiotherapy contributes to arm and breast edema. Lymphedema occurs most commonly in patients who have both axillary radiotherapy and surgery, is often triggered by a soft tissue infection, and is more common in obese patients. The incidence of arm edema is decreasing due to more conservative surgical treatment of the axilla and possibly due to more conservative management of the breast. Trends in breast edema are less discernible. Single-modality treatment of the axilla is associated with a very low incidence of arm edema. CONCLUSIONS: Lymphedema continues to be a problem in the care of the breast carcinoma patient. More conservative surgery combined with careful patient selection for nodal radiotherapy reduces its incidence. Radiotherapy technique, prompt treatment of soft tissue infections, and weight loss in obese patients each can contribute to prevention. The risk of lymphedema is greatly surpassed by the benefits of radiotherapy in the care of the breast carcinoma patient. PMID- 9874400 TI - The pathophysiology of lymphedema. AB - BACKGROUND: All edemas result from an imbalance between capillary filtration and tissue (lymph) drainage. This basic approach was adopted to investigate mechanisms for chronic arm edema following breast carcinoma treatment. METHODS: A review of causes of lymphedema is presented plus the traditional pathophysiology of breast carcinoma related lymphedema (postmastectomy edema; PME). A summary of recent research that explored capillary filtration as a surrogate for lymph flow in the steady state is presented. RESULTS: A reduced interstitial protein concentration (relative to plasma) argues against lymphatic obstruction. Evidence exists that total arm blood flow (in some patients) and vascular bed size are increased in PME. CONCLUSIONS: The primary insult to the axillary lymphatic system by surgery and radiotherapy presumably is the root cause of PME; however, there is strong evidence to suggest that hemodynamic factors are contributory to the chronic swelling. PMID- 9874401 TI - A review of measures of lymphedema. AB - BACKGROUND: Lymphedema usually is identified by patients, and rarely is it screened for routinely. Many assessments have been reported and have been used in evaluating a variety of treatments for lymphedema. METHODS: A review of the literature was undertaken. RESULTS: Five frequently used measures of lymphedema include circumferential measures of limbs at various points (usually at bony landmarks); volumetric measures using limb submersion in water; skin tonometry, in which soft tissue compression is quantified; imaging techniques to describe tissue characteristics as well as to quantify soft tissue swelling (magnetic resonance imaging and computerized tomography; and ultrasound with and without Doppler flow studies for volumetric measures. Circumferential measures with calculations designed to compute limb volumes and volumetric measures are used most frequently, but these have some difficulty with reliability. No significant effort has been made to develop a patient based questionnaire that describes the size as well as the impact of lymphedema on an individual's functional level. CONCLUSIONS: Existing physical measures of lymphedema are available that are easy to use, inexpensive, have limited reliability, and do not address the issue of functional impact. Imaging techniques may provide valuable qualitative and quantitative information in selected populations. PMID- 9874402 TI - Imaging techniques in the management and prevention of posttherapeutic upper limb edemas. AB - BACKGROUND: Upper limb edema remains the most frequent complication after treatments for breast carcinoma. Various imaging techniques can be used to prevent these complications, to manage them, and to diagnose the possible lymphonodal evolution that may underlie these events. In the present paper, these techniques are reviewed. METHODS: Based on clinical experience as well as on the data from the literature, these imaging techniques are presented, and their contributions are analyzed. RESULTS: The pre- and post-operative imaging and research techniques of the so-called sentinel nodes (using blue dye, and/or lymphoscintigraphy, and/or gamma probe) appear to be very promising for defining patients in whom axillary node clearance either might be avoided or is indicated. Lymphoscintigraphic investigations also can be used after surgery and/or radiation therapy to define patients who either are at risk to develop upper limb edema or present with latent edema. In patients with clinically obvious edema, even if it is limited, lymphoscintigraphic techniques can provide a morphologic and functional definition of the condition. Venous echo Doppler can be used when abnormalities of the venous return are suspected. In diagnosing the lymphonodal evolution of the disease, techniques like immunoscintigraphy, positron emission tomography (using 18-fluoro-2-deoxy-D-glucose), X-ray computed tomography, and nuclear magnetic resonance imaging can contribute. CONCLUSIONS: Various imaging techniques can be used to prevent and/or to manage the upper limb edema that may complicate the treatment(s) of the patients with breast carcinoma. PMID- 9874403 TI - Precipitating factors in lymphedema: myths and realities. AB - BACKGROUND: Lymphedema is an all too common occurrence following breast carcinoma therapy. Despite its prevalence, the predisposing factors to the development of this secondary form of lymphedema remain poorly understood. METHODS: Several studies have addressed these questions and are reviewed here. RESULTS: Treatment factors that appear to predispose to the late, subjective appearance of lymphedema include the extent of axillary surgery and exposure to high dose axillary radiotherapy, particularly when combined with surgical clearance of the axilla. Other pertinent patient factors may include the presence of hypertension and exposure to airline travel. Clinical features unrelated to the risk of lymphedema development include patient age; drug therapy; time interval to presentation, surgery, or radiotherapy to the breast; total dose of radiation; and menopausal status. The potential importance of concomitant venous abnormalities in these patients is worthy of consideration. CONCLUSIONS: Breast carcinoma-related secondary lymphedema is an important subjective and functional problem for affected patients. Additional research into the predisposing factors to this common problem is likely to foster enhanced patient education and to produce more efficacious measures to control this disease. PMID- 9874404 TI - Psychosocial aspects of upper extremity lymphedema in women treated for breast carcinoma. AB - BACKGROUND: Lymphedema of the upper extremity following breast carcinoma is highly distressing and disabling. METHODS: A review is presented of the literature on psychosocial aspects of lymphedema. RESULTS: Studies have shown that women who develop lymphedema exhibit higher levels of psychological, social, sexual, and functional morbidity than women with breast carcinoma who do not develop this complication. Women who have poor social support, pain, lymphedema in the dominant hand, and/or a passive and avoidant coping style report the highest levels of disability. CONCLUSIONS: Due to the morbidity of lymphedema once it develops, prevention and information given early are extremely important. However, the recommendations made to women sometimes involve considerable changes in lifestyle but are not based on solid, prospective evidence. In addition, how to best impart and time information about lymphedema pre- and postoperatively require investigation. In this paper, the authors discuss the psychological and functional impact of lymphedema and strategies for intervention and research to help women prevent the condition or enhance coping with it once it develops. PMID- 9874405 TI - Overview of treatment options and review of the current role and use of compression garments, intermittent pumps, and exercise in the management of lymphedema. AB - BACKGROUND: Lymphedema is a relatively frequent complication following the management of breast carcinoma. Numerous therapeutic interventions have been offered to treat this potentially disabling and disfiguring condition. Consensus has not been attained among oncologists, surgeons, psychiatrists, and physical therapists concerning the appropriate treatment of lymphedema. METHODS: This review provides an overview of those treatment regimens that have been used in the past and, in some instances, have gone on to provide the foundation for the most widely prescribed interventions currently employed for the management of upper extremity lymphedema following breast carcinoma treatment. The use of intermittent pneumatic compression pumps as a part of an integrated multidisciplinary treatment approach incorporating garments, exercises, and massage also is discussed. RESULTS: A review of available literature suggests that a variety of traditional and commonly available techniques, when used appropriately in a multidisciplinary fashion, may lessen the cosmetic and physical impairments associated with acquired lymphedema. The role of surgery is unclear. Pharmacotherapies are a promising adjunct to manual and mechanical therapies. CONCLUSIONS: The appropriate use of readily available treatment approaches may lessen the severity of acquired lymphedema following breast carcinoma therapy. A comprehensive therapeutic approach should be employed in the management of lymphedema, including attention to the functional, cosmetic, and emotional sequelae of this potentially disabling condition. To that end, a recommendation for a comprehensive treatment regimen is provided. PMID- 9874406 TI - Conservative approaches to lymphedema treatment. AB - BACKGROUND: Upper extremity lymphedema can develop after surgery for breast carcinoma. Once developed, it becomes a chronic problem that women must cope with for the rest of their lives. Steps to prevent lymphedema should begin immediately after surgery. However, there is little information available about what actually causes lymphedema; therefore, it is difficult to prevent, and there is some controversy over how women should be treated once lymphedema has developed. METHODS: The literature was reviewed to understand the education about arm care provided to women during and after the short hospital stay for breast carcinoma surgery. Evaluation and treatment options for lymphedema and complications resulting from lymphedema were explored. RESULTS: Women are provided with basic arm care information after surgery; however, many women require reinforcement from health professionals, such as physical or occupational therapists, to reach optimum functional outcomes. If lymphedema does develop, then there are two treatment regimes that have been used. The compression pump, along with skin care, exercise, and compression garments, is one option. However, there is little consistency with the length of time the pump should be used or the optimum number of days required to receive the best results. The second treatment option is known as complex decongestive physiotherapy or complex physical therapy. Arm care, therapeutic exercises, manual lymph drainage, and compression bandages and/or garments comprise this treatment regime. Decreases in lymphedema are noted if women are compliant with the prescribed treatment program. CONCLUSIONS: Women must be educated about possible complications after breast surgery. This should be a team effort, with physicians, nurses, physical and occupational therapists, and Reach to Recovery volunteers from the American Cancer Society all participating in the process. If women do develop lymphedema, then an individual treatment program must be established, and adherence to the program must be stressed. More research is needed to determine the optimum treatment regime for women who develop lymphedema. PMID- 9874407 TI - The treatment of lymphedema. AB - BACKGROUND: Before the treatment of arm lymphedema after breast carcinoma treatment with complex decongestive physiotherapy can be initiated, it is mandatory to differentiate between benign and malignant forms (due to relapse) and to establish the diagnosis of accompanying diseases, if present. METHODS: In benign lymphedemas, the aim of complex decongestive physiotherapy is to restore the symptom free "Stage 0 of latency" and to maintain fitness for work. The palliative treatment of malignant lymphedemas results in the amelioration of the quality of life. RESULTS: The results of treatment depend on the experience of the physician in clinical lymphology, on the training and dedication of the lymphedema therapist, and on the compliance of the patient. CONCLUSIONS: A study concerning gene expression has shown that complex decongestive physiotherapy influences the pathological alterations of the interstitium in lymphedema patients. PMID- 9874408 TI - The physical treatment of upper limb edema. AB - BACKGROUND: Edema of the upper limb, without any doubt, constitutes the most invalidating complication of breast carcinoma treatment. The swelling of the limb results from decreased liquid evacuation by surgical intervention at the axillary level and also by the eventual treatment by cobaltotherapy. METHOD: The physical treatment for edema of the limb consists of a combination of therapies that were tested for their effectiveness in laboratories on healthy students and also on patients who underwent surgery for breast carcinoma. The treatment consists of the application of manual lymphatic drainage (type Leduc), the use of multilayered bandages, and the use of intermittent pneumatic compression. The population studied was represented by 220 patients who underwent breast surgery. The authors followed their evolution during the first 2 weeks of treatment. Patients were not hospitalized. The edema was measured by using marks tattooed on the skin. RESULTS: The limb that developed edema was compared with the healthy limb. The most important reduction was obtained in the first week. The decrease was equivalent to 50% of the average of the difference between both upper limbs. During the second week, the results obtained stabilized; however, there was a slight decrease at the end of the second week. CONCLUSIONS: The physical treatment of edema represents the preferred therapeutic approach. However, it must answer to well-defined criteria to be efficient and for long-lasting effects. The physical treatment is used to treat outpatients, allowing them to follow a normal lifestyle. PMID- 9874409 TI - The Vodder School: the Vodder method. AB - BACKGROUND: The history and development of manual lymphatic drainage (MLD) from Winiwarter to Vodder and the Vodder School of today are discussed. METHODS: The Vodder technique differs in the use of adapted pressure and its application. The constant change in pressure optimizes results, moving fluid in the skin, increasing lymphomotoricity, and softening fibrosis, with the positive side effects of reducing pain and relaxing tense muscles. Another difference from other methods is the technique of stretching skin, not sliding it. Because of the fluid content in lymphedema, which is different from all other edemas, the combination of MLD with compression treatment is the only solution for this pathology. Depending of it severity, each case requires individualized treatment. Phase 1 (intensive treatment) consists of daily treatment with up to two sessions per day for up to 2 hours. This phase is combined with special, individual skin care and remedial exercise. In phase 2, the goal of treatment is to maintain the results achieved in phase 1. The frequency of treatment is changed, but there is still the need for permanent, continued therapy. RESULTS: In phase 1, an average reduction of more than 40% of edema volume is achieved. In phase 2, the results are maintained and, with repetitions of phase 1, further improvement is possible. Thus, long term results with permanent improvement are possible. CONCLUSIONS: Because of the complexity of the technique, no one can learn MLD in 1 week. Students require a great deal of correction, and the technique must be checked constantly. To become a certified Vodder therapy, a 4-week education program must be completed, and reviews must be attended every 2 years to maintain certification. The best education produces the best results for patients as long as patients are compliant. Therefore, the Vodder School also includes a patient education program as part of its curriculum. PMID- 9874410 TI - Treatment for lymphedema of the arm--the Casley-Smith method: a noninvasive method produces continued reduction. AB - BACKGROUND: This paper gives an outline of the Casley-Smith method for the treatment of lymphedema of the arm. It includes a brief summary of the development of manual techniques and the terminology applied to them. METHODS: The four principles of this method are skin care, manual lymphatic drainage, compression in the form of bandaging and/or garments, and exercise. The massage techniques, especially where they differ from other schools, are described in some detail, as are the principles that apply in compression and maintenance of reduction in lymphedema. RESULTS: The results of this method have been analyzed both in Australia and in the United States and are discussed briefly. Mention is made of the benefits of the benzopyrones, which have been used for many years, when added to the above treatment. Both benzopyrones and exercise will produce a continued reduction after the treatment course. They are particularly useful in a less compliant patient. It is stressed that the effect of patient compliance, particularly after treatment, makes a great difference to the ongoing success of the regime. CONCLUSIONS: A comparison is drawn between the efficacy of various current treatments and their cost. This shows that this combined and conservative method of treatment should be considered before recourse to pumps or surgery. The latter seldom achieve the results of decongestive lymphatic drainage, and, in the long term, they are more expensive. Certain preventive measures may be indicated following, e.g., mastectomies. Prevention of the onset of lymphedema is of extreme importance. However, a return to as normal a lifestyle as possible by the patient is also essential. The earlier treatment begins after the onset of lymphedema, the better the prognosis for the patient. Lymphedema can and should be treated. PMID- 9874411 TI - Complete decongestive physiotherapy and the Lerner Lymphedema Services Academy of Lymphatic Studies (the Lerner School). AB - BACKGROUND: Lymphedema is more common than most physicians realize. It is also an incurable, life long condition that has never been treated effectively in the past in our country. METHODS: The author describes complete decongestive physiotherapy (CDP) and his contribution to making it available all across the United States. This was done by establishing a network of outpatient CDP clinics where patients can be treated effectively and a training school, the Lerner Lymphedema Services Academy of Lymphatic Studies (the Lerner School), where physicians and therapists can be trained in all facets of the lymphedema problem and where the CDP method with all of its components is taught. RESULTS AND CONCLUSIONS: The superiority of CDP to pneumatic pumps and to surgical procedures used to treat lymphedema is discussed. A description of the Lerner School's philosophy and curriculum is included. PMID- 9874412 TI - Current status of education and treatment resources for lymphedema. AB - BACKGROUND: Secondary lymphedema (LE) resulting from breast cancer surgery has continued to be an ignored medical diagnosis in U.S. medicine. Subsequently, the majority of women/men undergoing axillary lymph node dissection have not received education in pre- or postoperative LE prevention, causing hundreds of thousands of patients to develop LE. As a result of this ignorance, once the LE develops, these patients receive no or harmful treatment. METHODS: Over the last decade, the National Lymphedema Network (NLN) has created awareness in the medical community, among patients, and in the general public through information dissemination, educational materials, a national conference, and the activism of patients and a small group of concerned professionals. Complete decongestive physiotherapy (CDP), which includes manual lymph drainage, is a successful treatment for LE that has now been introduced. RESULTS: Slowly, the U.S. medical community is beginning to recognize and support this condition and its treatment, and secondary LE has now become an acceptable diagnosis (ICD9-457.0). Subsequently, due to patient and medical activism and the work of the NLN, Medicare now covers 2 weeks of CDP treatment (in Florida), and two landmark bills (AB-12 and S-609) that will cover LE treatment for postbreast carcinoma patients are currently pending (as of February 22, 1998). CONCLUSIONS: In the last decade, the NLN has developed a strong foundation of knowledge and has planted LE on the medical map in the United States. With collaboration between the NLN and all breast carcinoma groups and LE specialists, it is hoped that lymphology will become a standard course of study in medical schools across the country and that LE will become a household word, with proper treatment available for postbreast carcinoma LE patients in the United States. PMID- 9874413 TI - Lymphedema management training for physical therapy students in the United States. AB - BACKGROUND: The objective of this study was to determine to what extent accredited physical therapy programs in the United States were presenting the principles of lymphedema management and whether regional differences existed. METHODS: States were grouped into four geographic regions: Northeast, South, Midwest, and West. From mid-June to mid-July, 1997, 63 of 148 (42.6%) accredited physical therapy (PT) programs in the United States completed and returned the questionnaires. Participants received a cover letter, consent form, and lymphedema survey by e-mail, facsimile, or regular post. The lymphedema survey covered a wide variety of issues relating to five areas: 1) general and 2) specific anatomy and physiology of the lymphatic system, 3) pathogenesis of lymphedema, 4) traditional (compression pumps/garments), and 5) innovative (European/Australian) treatment techniques for lymphedema. "Yes" responses indicated that specific information was included in the curriculum. Frequency of yes responses for each of the five areas were counted and converted into percentages. Regional responses were compared with the total combined responses with a modified binomial technique. RESULTS: PT programs in the United States were providing 89% of our designated content in the general anatomy and physiology of the lymphatic system, 73% in the pathogenesis of lymphedema, 65% in traditional treatment techniques, 48% in innovative treatment techniques, and 42% in the specific anatomy and physiology of the lymphatic system. No individual region differed significantly (P > 0.05) from the combined results. CONCLUSIONS: The participating PT programs appeared to be providing instruction in general anatomy and physiology of the lymphatic system, pathogenesis of lymphedema, and traditional treatment techniques. However, far less instruction on the specific anatomy and physiology of the lymphatic system and innovative treatment techniques is offered. We believe that PT students would benefit with more curricular content in these latter two categories in order to acquire the knowledge and skills to combat the devastating effects of lymphedema. PMID- 9874414 TI - Lymphedema: patient and provider education: current status and future trends. AB - BACKGROUND: The majority of patients schedules for breast surgery or radiation therapy for breast carcinoma do not receive basic information about the risk of lymphedema or its treatment. METHODS: A review of the literature documenting patient and provider education on the risk, prevention and treatment of lymphedema was undertaken. RESULTS: Formal education about lymphedema is not a part of the training of most medical and allied health professionals. Thus, patients do not receive information about the risk, prevention, or treatment of lymphedema. CONCLUSIONS: Educational strategies for patients and providers need to be developed and implemented. PMID- 9874415 TI - American Cancer Society Lymphedema Workshop. Workgroup I: Treatment of the axilla with surgery and radiation--preoperative and postoperative risk assessment. PMID- 9874416 TI - American Cancer Society Lymphedema Workshop. Workgroup II: Patient education--pre and posttreatment. PMID- 9874417 TI - American Cancer Society Lymphedema Workshop. Workgroup III: Diagnosis and management of lymphedema. PMID- 9874419 TI - American Cancer Society Lymphedema Workshop: Workgroup V: Collaboration and advocacy. PMID- 9874418 TI - American Cancer Society Lymphedema Workshop. Workgroup IV: Lymphedema treatment resources--professional education and availability of patient services. PMID- 9874420 TI - Verrucous carcinoma, then and now. AB - The salient clinical and pathological features of verrucous carcinoma, as first described by Ackerman in 1948, are reviewed. Two case reports are presented. The first concerns a personal experience with a patient who had five surgical procedures for multifocal oral verrucous carcinoma and remains alive and well 28 years after her initial treatment. The second report highlights the remarkable story of President Grover Cleveland's secret surgery for verrucous carcinoma of the palate in 1893. Current concepts about the management of this uncommon tumor are reviewed, as well as some recent investigations that deal with its etiology. PMID- 9874421 TI - Hayes Martin lecture. The making of a specialty. PMID- 9874423 TI - 99m-Technetium sestamibi localized solitary parathyroid adenoma as an indication for limited unilateral surgical exploration. AB - BACKGROUND: Because of its successful localization of solitary adenomas, 99m Technetium sestamibi (MIBI) may challenge the standard operation for primary hyperparathyroidism. METHODS: Thirty-five consecutive patients underwent preoperative MIBI localization to optimize a surgical approach. Single-site localization in 21 patients directed a limited unilateral neck exploration (UNE) with adenomectomy and ipsilateral gland biopsy. Fourteen patients who did not localize underwent bilateral neck exploration (BNE). Conversion to a bilateral operation was required in 1 UNE patient because no adenoma was found on that side. RESULTS: There were no significant differences in preoperative and postoperative serological markers between the two groups. However, the total operative time for UNE (49 +/- 21 minutes) was significantly less than for BNE (103 +/- 45 minutes; P <0.001). CONCLUSIONS: Preoperative MIBI scan-directed limited unilateral neck operation may be used reliably for primary hyperparathyroidism due to a single adenoma, and thereby reduce operative time, extent of surgical dissection, and risk. PMID- 9874422 TI - Efficient gene transfer to human squamous cell carcinomas by the herpes simplex virus type 1 amplicon vector. AB - BACKGROUND: This study evaluates the efficiency of herpes simplex virus (HSV) mediated gene transfer in human squamous cell carcinoma (SCC) cell lines in vitro and in vivo when delivered by selective intra-arterial perfusion. METHODS: Human head and neck SCC were exposed to HSV-LacZ and HSV-interleukin-2 (IL-2) and gene transfer and expression assessed by X-gal staining and enzyme-linked immunosorbent assay, respectively. Hamster cheek pouch tumors were perfused with HSV-LacZ or HSV-IL-2, by microcannulating the external carotid artery, and gene transfer determined. RESULTS: A ratio of 5 viral particles per tumor cell achieved gene transfer rates exceeding 50%. Interleukin-2 levels of 287 +/- 17 to 424 +/- 8.4 ng per million cells were achieved at a ratio of 2 viral particles per tumor cell. Selective intra-arterial perfusion of the HSV-IL-2 vector yielded IL-2 levels of 45.8 +/- 17.0 pg per g tumor. CONCLUSIONS: HSV amplicon vectors are efficient vehicles for gene transfer in vitro in human head and neck SCC cell lines and in vivo when introduced by selective intra-arterial perfusion. PMID- 9874424 TI - The role of fine-needle aspiration biopsy and flow cytometry in the evaluation of persistent neck adenopathy. AB - BACKGROUND: The role of fine-needle aspiration biopsy (FNAB) and flow cytometry (FCM) in the evaluation of persistent neck adenopathy has not been completely defined. METHODS: All patients who underwent FNAB of an abnormal lymph node from 1990 to 1997 were identified, and the results of FNAB and FCM were correlated with histology or clinical follow-up. RESULTS: FNAB was performed on 133 patients with an abnormal lymph node in the neck. FNAB was malignant in 82 (62%), atypical in 13 (10%), benign in 27 (20%), and nondiagnostic in 11 (8%) patients. FNAB was compared with histologic results in 114 (86%) or with the results of clinical follow-up (average 17 months) in 12 patients (9%). Seven patients (5%) were lost to follow-up. Lymphoma and metastatic carcinoma were diagnosed in 25 (19%) and 76 (57%) patients, respectively. Sensitivity of FNAB and FCM was 96% and 100%, respectively, and specificity was 58% and 100%, respectively. Patients with Hodgkin's lymphoma accounted for 80% of false negative FCM results. FCM was nondiagnostic in 3 of 34 patients (9%). Definitive subclassification of lymphoma was accomplished in only 3 of 17 (18%) patients in whom FCM was performed. CONCLUSIONS: FNAB is reliable in identifying metastatic disease, non-Hodgkin's lymphoma, and reactive lymphadenopathy in the neck. FNAB and FCM still have a limited role in definitive diagnosis of Hodgkin's lymphoma and classification of non-Hodgkin's lymphoma, underscoring the importance of excisional biopsy. PMID- 9874425 TI - Donor-specific tolerance induction in composite tissue allografts. AB - BACKGROUND: Although prolonged composite tissue allograft (CTA) survival is achievable in animals using immunosuppressive drugs, long-term immunosuppression of CTAs in the clinical setting would be unacceptable for most patients. The purpose of this study was to develop a model for reliable CTA tolerance induction in the adult rat across a strongly antigenic MHC mismatch without the need for long-term immunosuppression. METHODS: Chimeras were prepared using rat strains with strong MHC incompatibility [WF (RT1Au) + ACI (RT1Aa) --> WF, n = 13]. Syngeneic (WF) and allogeneic (ACI) bone marrow (BM) was harvested and T-cell depleted. Following confirmation of T-cell depletion by flow cytometry, a mixture of T-cell depleted syngeneic and allogeneic BM was injected into the recipient animals (all recipients pretreated with low-dose irradiation, 500 to 700 cGy). In addition, the recipient animals received a single dose of ALS (10 mg) 5 days prior to bone marrow transplantation (BMT) and tacrolimus (1 mg/kg/day) from the day prior to BMT to 10 days postoperatively. Rat chimeras were characterized by flow cytometry at 3 and 12 months after BM reconstitution and following hindlimb transplantation. RESULTS: Peripheral blood lymphocyte chimerism (WF/ACI) remained stable >12 months after BM reconstitution in 10 of 13 animals. Multilineage chimerism of both lymphoid and myeloid lineages was present, suggesting that engraftment of the pluripotent rat stem cell had occurred. In animals with donor chimerism >60%, there was no sign of limb rejection for the duration of the study. All animals with chimerism <20% developed moderate signs of rejection clinically and histologically. Gross motor and sensory reinnervation (weight bearing, toe spread) occurred at >60 days in 6 of 9 rats. Postoperative flow cytometry studies demonstrated stable chimerism in all animals studied (n = 7). CONCLUSIONS: Stable mixed allogeneic chimerism can be achieved in a rat hindlimb model of composite tissue allotransplantation. Hindlimb allografts to mixed allogeneic chimeras exhibit prolonged, rejection-free survival. This represents the first reliable model demonstrating rejection-free CTA survival in an adult animal without the long-term use of immunosuppressive agents across a strongly antigenic MHC mismatch. PMID- 9874426 TI - Results of a prospective trial on elective modified radical classical versus supraomohyoid neck dissection in the management of oral squamous carcinoma. Brazilian Head and Neck Cancer Study Group. AB - BACKGROUND: Elective treatment of the neck in oral squamous cell carcinoma has changed over the last 20 years. The main object of this report is to present the results of a multi-institutional prospective study designed to compare standard treatment with modified radical classical neck dissection (MRND) to supraomohyoid neck dissection (SOH) in the management of the clinically negative neck in oral cancer patients. PATIENTS AND METHODS: A total of 148 patients were included in the trial. All patients had previously untreated T2 to T4 N0 squamous cell carcinoma of the oral tongue (62 cases), floor of the mouth (49 cases), inferior gingiva (12 cases), or retromolar trigone (25 cases). Tumor stages were T2, 91; T3, 27; and T4, 30. There were no significant imbalances between groups. RESULTS: The false-negative rate was 28%, and most positive nodes were sited at level II and III. Complications were seen in 41% of MRND patients and in 25% of SOH patients (P = 0.043). Median total duration of hospitalization was 9 days in MRND patients and 7 days in the SOH group. To date, 19 and 16 patients presented with local and neck recurrences, respectively. The 60-month actuarial survival rates were 63% in the MRND group and 67% in the SOH group (P = 0.7150). CONCLUSIONS: This study demonstrates that the recurrence and survival rates were similar in both groups. SOH neck dissection can be recommended as standard elective treatment for patients with T2-T4 oral squamous cell carcinomas. PMID- 9874427 TI - Low prevalence of human papillomavirus in a geographic region with a high incidence of head and neck cancer. AB - BACKGROUND: The main purpose of this study was to determine the prevalence of human papillomavirus (HPV) infection in patients with head and neck carcinomas from Brazil. MATERIALS AND METHODS: Forty-five patients with head and neck squamous cell carcinoma were included in the study, from 1995 to 1996. Forty-two were male and 3 female, with age ranging from 32 to 82 years (median 61). Five patients (11%) did not have previous history of use of tobacco and 38 (90.5%) were heavy smokers. Tumor sites were pyriform sinus, 10; tongue, 11 (oral, 6; base, 5); larynx, 7; floor of mouth, 3; tonsil, 6; retromolar area, 3; inferior gingiva 2; buccal mucosa, 2; and maxillary sinus in 1 patient. Twenty-five were stage IV, 17 stage III, and 3 stage II. RESULTS: The presence of HPV DNA was detected in 5 of 45 patients (11%), all of them with HPV 16. Two patients had HPV DNA in normal mucosa and tumor tissue, 1 patient had HPV DNA only in the normal mucosa and tumor tissue, 1 patient had HPV DNA only in the normal mucosa, and 2 patients were positive for HPV DNA in tumor tissue. Four patients were male and 1 was female; 2 patients were nonsmokers. Three patients had tonsil carcinoma, 1 patient had a tongue carcinoma, and 1 patient had a pyriform sinus cancer. CONCLUSIONS: The role of chemical carcinogens seems to be more important in the genesis of head and neck cancer than is HPV infection. The presence of HPV DNA in 5 of 45 patients stimulates further investigation to determine the role of HPV as a risk factor for head and neck carcinoma. PMID- 9874429 TI - Resection of the metatarsal head for diabetic foot ulcers. AB - BACKGROUND: Diabetic foot ulceration is a worldwide health problem. Approximately 15% of the 10 million diabetic patients in the United States will develop a foot ulceration at some time in their lives. The presence of a foot ulcer in this population is extremely debilitating and dramatically increases the risk of lower extremity amputation, accounting for approximately 67,000 lost limbs each year. Additionally, the costs associated with treating foot ulcers in diabetic patients is a major expense in the overall care of this patient group. METHODS: An 11-year retrospective study was conducted to evaluate 101 consecutive patients with diabetic ulcers of the forefoot who were treated using resection of the metatarsal head as the primary means of obtaining wound closure. RESULTS: The results indicate that 88% of the ulcers were healed by using this technique, and relatively more rapidly than would be expected when compared with historical norms. CONCLUSIONS: Resection of the metatarsal head is a safe and relatively inexpensive procedure that facilitates closure of the lesion, helps to control infection, and prevents countless and costly amputations. PMID- 9874428 TI - Predictive factors for functional recovery after free tissue transfer oromandibular reconstruction. AB - BACKGROUND: Recovery of upper aerodigestive tract function after reconstruction of segmental oromandiblectomy defects is frequently incomplete. The purpose of this study was to quantitate postreconstruction function and define variables that predict functional outcome in this population. METHODS: A prospective study of 21 patients who underwent microvascular free tissue transfer reconstruction of segmental oromandibular defects was performed. Measures of swallowing, speech, bite, and oral intake were performed preoperatively and at 1, 3, 6, and 12 months postoperatively or until plateau. Preoperative versus maximal postoperative measures were compared and correlated with nine potentially predictive variables. Univariate and multivariate analyses were performed to determine the most significant predictive factors. RESULTS: Baseline function in the study population was abnormal. Postoperative bite force improved, but swallowing, speech, and oral intake were worse than preoperative. Significant (univariate) predictors of outcome included diagnosis of cancer, tongue resection, pharynx resection, and flap skin paddle area. Only tongue resection remained significant in multivariate analysis. CONCLUSIONS: Increasing need for oropharyngeal lining replacement, especially after tongue resection, is the most important predictor of functional outcome in reconstruction of segmental mandible defects. PMID- 9874430 TI - Tumor vascularity predicts recurrence in differentiated thyroid carcinoma. AB - BACKGROUND: New prognosticators are necessary for optimizing the extent of thyroidectomy and adjuvant radiotherapy in differentiated thyroid carcinoma. METHODS: Tumor microvessel density (MVD), expression of thymidine phosphorylase (dThdPase) and CD68 stained macrophages were evaluated in 71 differentiated thyroid carcinomas by an immunohistochemical method. RESULTS: The recurrence-free survival was significantly (P <0.05) shorter in patients with hypervascular tumors but was not related to the dThdPase expression. Significant (R = 0.323, P <0.001) correlation between dThdPase expression score and increment in MVD was documented. Frequent high MVD and dThdPase expression were encountered in tumors >3 cm. Twenty-two of the 27 (82%) patients expressing high dThdPase were also evaluated positive for CD68 (P <0.001). CONCLUSIONS: Tumor MVD might be a new prognostic indicator of differentiated thyroid carcinoma, whereas dThdpase expression with high MVD might serve to identify a subgroup of thyroid carcinoma patients as potential candidates for adjuvant radiotherapy. PMID- 9874431 TI - Management of the clinically positive neck in organ preservation for advanced head and neck cancer. AB - BACKGROUND: To investigate clinicopathologic predictive criteria for the optimal management of neck metastases in patients with advanced head and neck cancers treated with combined chemoradiotherapy. METHODS: Prospective study, 48 patients. Mean length follow-up, 23 months. RESULTS: Neck stage predicted neck response to chemoradiotherapy; N3 necks showed more partial responses (P = 0.04), and N1 necks showed more complete responses (P = 0.12). Primary tumor site strongly predicted the pathologic response found on neck dissection in patients with a clinical partial response (cPR) following chemoradiotherapy. There was no difference in survival between patients with a clinical complete response (cCR) after chemoradiotherapy, and patients with a pathologic complete response (pCR) after neck dissection (P = 0.20); however, when grouped together, these patients survived longer than did patients with a pPR at neck dissection (P = 0.06). CONCLUSIONS: Clinical response to induction chemotherapy is a poor predictor of ultimate neck control. Induction chemotherapy followed by chemoradiotherapy, and planned neck dissection for patients with persistent cervical lymphadenopathy, provides good regional control. PMID- 9874433 TI - The effects of hyperbaric oxygen on normal and ischemic colon anastomoses. AB - BACKGROUND: Leakage from colonic anastomoses is a major complication causing increased mortality and morbidity, and ischemia is a well-known cause of this event. Inadequate tissue oxygenation could be reversed by using hyperbaric oxygen. This study was designed to investigate the effects of hyperbaric oxygen on the healing of ischemic and normal colon anastomoses in the rat model. METHODS: Standardized left colon resection 3 cm above the peritoneal reflection and colonic anastomosis were performed in 40 Wistar rats divided into four groups. The control group (I) received no further treatment. To mimic ischemia, 2 cm mesocolon was ligated on either site of the anastomosis in group II and IV rats. Hyperbaric oxygen therapy was started immediately after surgery in group III and IV rats (therapeutic groups). All animals were sacrificed on the fourth postoperative day. After careful relaparotomy, in situ bursting pressure was measured. The hydroxyproline contents of the anastomotic segments in equal length were determined. RESULTS: The hydroxyproline assay revealed that rats in group II with ischemic colonic anastomosis have significantly lower levels (P <0.05). The highest levels are in the group III rats with normal colonic anastomosis treated by hyperbaric oxygen (P <0.05). There was no significant difference in hydroxyproline levels between group II and group IV animals (P >0.05). Group III animals had significantly higher bursting pressures than any other group (P <0.05). Group II rats had lowest bursting pressures (P <0.05). Group IV animals had significantly higher levels than group II (P <0.05). Mean bursting pressure values both in groups III and IV and hydroxyproline levels in group III were significantly increased by hyperbaric oxygen therapy (P <0.05). CONCLUSIONS: Ischemia impairs anastomotic healing. Hyperbaric oxygen increases anastomotic healing of both normal and ischemic colonic anastomosis and reverses ischemic damage. This study demonstrated that hyperbaric oxygen improves anastomotic healing. PMID- 9874432 TI - A randomized, double-blind trial for stress ulcer prophylaxis shows no evidence of increased pneumonia. AB - BACKGROUND: H2-receptor antagonists are commonly used for stress ulcer prophylaxis on intensive care units. However, there is evidence that via the route of an elevated gastric pH, followed by bacterial overgrowth and subsequent tracheal aspiration, pneumonia could occur. In line with this assumption total gastrectomized patients should develop a very high incidence of pneumonia, which is actually not the case. We therefore formulated the hypothesis that stress ulcer prophylaxis with H2-receptor antagonists does not lead to an increased pneumonia rate. METHODS: A total of 158 patients with mechanical ventilation > or =48 hours of a surgical intensive care unit were randomized to the following groups: A, placebo (n = 57); B, pirenzepine (3 x 10 mg intravenously, n = 44); and C, ranitidine (3 x 50 mg intravenously, n = 57). RESULTS: The pneumonia rate in ranitidine-, pirenzepine-, and placebo-treated patients is 10 of 57, 10 of 44, and 12 of 57, respectively. CONCLUSIONS: Pneumonia rate is not adversely affected by H2-receptor antagonists in stress ulcer prophylaxis. PMID- 9874434 TI - Randomized trial comparing packed red cell blood transfusion with and without leukocyte depletion for gastrointestinal surgery. AB - BACKGROUND: Allogeneic transfusion is associated with postoperative infections that significantly prolong hospital stays and increase costs. Recent studies suggest that filtering leukocytes from blood prior to transfusion reduces the risk of postoperative infection associated with blood transfusion. We compared the incidence of postoperative infections, hospital stays, and hospital charges of gastrointestinal surgery patients transfused with packed red cells or leukocyte-depleted cells. METHODS: Consecutive patients admitted for elective gastrointestinal surgery without previous blood transfusion were randomized to receive routine packed red cells or packed red cells filtered to remove leukocytes if transfusion was required. Multivariate analysis was used to assess the significance of the relationship between leukocyte-depleted blood and postoperative infectious complications, postoperative stay, and hospital charges. RESULTS: Fifty-nine (27%) of the 221 patients were transfused. The most significant variable related to transfusion was intraoperative blood loss (P <0.0001), followed by admission hematocrit (P <0.0001) and age (P = 0.0022). Infections were noted in 16% of the patients: 11% of untransfused patients, 16% of leukocyte-depleted blood recipients, and 44% of patients transfused with packed red cells. Both operative site and nosocomial infections were significantly (P <0.001) more frequent in patients transfused with packed red cells compared with patients transfused with leukocyte-depleted red cells. Postoperative stays averaged 9 days for untransfused patients, 12 days for leukocyte-depleted recipients, and 18 days for recipients of packed red cells. Hospital charges were $19,132, $33,954, and $41,002, respectively. Both transfusion and infection were significantly (P <0.001) related to postoperative stay in multivariate analysis. Hospital charges were significantly related to postoperative stay (P <0.001), blood loss (P <0.001), age (P <0.001), infection (P = 0.007), and randomization to packed red cells (P = 0.032). CONCLUSIONS: Filtering blood of leukocytes prior to transfusion for elective gastrointestinal surgery is associated with lower risk of postoperative infection, shorter postoperative stays, and lower hospital charges. PMID- 9874435 TI - Comparison of relapse and long-term survival between pylorus-preserving and Whipple pancreaticoduodenectomy in periampullary cancer. AB - BACKGROUND: Whether long-term survival and type of recurrence in periampullary cancer are influenced by the type of surgical procedure or not is still obscure. The purpose of this review was to analyze the results at a single institution. METHODS: Between 1982 and 1996, 113 consecutive patients with either distal bile duct or ampullary cancers underwent either pylorus-preserving pancreaticoduodenectomy (PPPD) or the Whipple procedure. The median follow-up was 30 months (range 12 to 150). Patient records were reviewed to determine type of recurrence and survival. RESULTS: In distal bile duct or ampullary cancers, comparison of actuarial survival curves between PPPD and the Whipple operation revealed no significant differences. Even in node-positive patients, the 5-year survival rates did not differ significantly between those undergoing PPPD or the Whipple operation. Tumor recurrence was not correlated with the operative method, although among types of tumor recurrence, the rate of blood-borne metastasis was higher in distal bile duct or ampullary cancers. CONCLUSIONS: PPPD is an acceptable alternative to the Whipple procedure in the treatment of periampullary cancer. Long-term survival and type of recurrence were not influenced by these types of surgical procedures. PMID- 9874436 TI - Treatment of chronic thoracic duct fistula with the sternocleidomastoid muscle flap. AB - Thoracic duct fistula is a rare but potentially serious complication of head and neck surgery. The fistulae can be difficult to treat, and several techniques, both operative and nonoperative, are advocated. We present a simple technique for the treatment of chronic thoracic duct fistula where other procedures have failed. After identification and ligation of the duct, the area is covered with the clavicular head of the sternocleidomastoid muscle. PMID- 9874437 TI - Intestinal anastomosis with the biofragmentable anastomosis ring. AB - BACKGROUND: The fear of anastomotic complications prevents the spread of the use of the biofragmentable anastomosis ring (BAR) in intestinal surgery. PATIENTS AND METHODS: A total of 453 patients underwent intestinal resection and anastomosis with the BAR. RESULTS: In all, 514 anastomoses have been carried out, 424 (83%) in elective settings, and 90 (18%) in emergency. Fifty-one patients had multiple anastomoses. Reoperation was performed in 4 patients (1%) who had a complete anastomotic leakage. In 13 patients (3%), anastomotic leakage was partial, and only 1 patient required reoperation. No postoperative intestinal obstruction occurred. Four patients (1%) developed late anastomotic strictures, which have been treated by endoscopic dilation. CONCLUSIONS: The results of our experience and those of other large clinical series definitely confirm the effectiveness of the BAR method, which seems to be a standard, easy, rapid, and safe technique either in elective or emergency surgery. PMID- 9874438 TI - A solution to the problem of undictated operative reports by residents. AB - PURPOSE: The objective of this study was to reduce the number of operative reports assigned to the surgical residents but not dictated within 48 hours by 80% within 6 months. METHODS: A before-after trial was conducted in a teaching hospital (part of a major academic medical center) in an urban setting, of a complex intervention based on a theoretical framework for behavioral change. Data were collected for more than 6 months before the intervention, and then in three separate time periods during the next 18 months. The intervention, lasting 41 weeks, and directed specifically at undictated operative reports, comprised the following: educational sessions, posting of residents' names with delinquency rates, attending/resident interactions, telephone reminders, rewards, and punishments. The population studied comprised every resident who rotated through the general surgery service over a 2-year period 1995 to 1997. RESULTS: The mean (+/-SD) number of undictated operative reports declined progressively from 72 (+/ 8.3) to 6 (+/-2.6). This 92% reduction was statistically significant (P <0.00005). The mean number of undictated discharge summaries declined progressively from 54 (+/-11.2) to 13 (+/-8.1). This 76% reduction was also statistically significant (P <0.00005). There was a close correlation between the operative reports and the discharge summaries (r = 0.82). CONCLUSIONS: The performance of residents on the surgical service in respect to operative report dictation can be profoundly influenced by a carefully targeted set of interventions based on behavioral theory. The improved performance can be maintained with simple reminders and the halo effect of the intervention extends to the dictation of discharge summaries. PMID- 9874439 TI - Opinions of practicing general surgeons on surgical education. AB - BACKGROUND: Collective opinions of practicing general surgeons on the current state of general surgical resident education are unknown. METHODS: A 26-item survey was mailed to practicing general surgeons in Minnesota and Texas. Average scores on 17 Likert-formatted questions and opinions on multiple-choice questions were compared by geographic area and academic affiliation. RESULTS: Overall response was 954 of 1,745 (55%). All surgeons felt changes were needed in surgical education. There was agreement by geographic area and academic affiliation that the current system of resident education allows chief residents to graduate with significant gaps in their education, and that the responsibility for correcting these gaps lies with the residency program. CONCLUSIONS: Opinions of general surgeons in two geographic areas and of differing academic affiliation regarding surgical education showed marked similarity. These data suggest change in the process of surgical education is the responsibility of the residency program and should be a priority for the profession. PMID- 9874440 TI - Surgical Internet at a glance: volume VI. PMID- 9874441 TI - Can a one-day workshop alter surgeons' teaching practices. PMID- 9874442 TI - "Multiple primary carcinomas of the large intestine"--50 years later. PMID- 9874443 TI - Economic analysis of expensive technologies: the case of erythropoietin. PMID- 9874444 TI - Experience enhances mammographic interpretation, doesn't it? PMID- 9874445 TI - Experience enhances mammographic interpretation, doesn't it? Yes, it does! PMID- 9874446 TI - The National Cancer Data Base Report on thyroid carcinoma: reflections of practice patterns. PMID- 9874447 TI - Malignant mesothelioma of the tunica vaginalis testis: review of the literature and assessment of prognostic parameters. AB - BACKGROUND: Only 73 cases of malignant mesothelioma of the tunica vaginalis testis have been reported in the last 30 years. Although these tumors were most often seen in patients between ages 55 and 75 years, 10% of the patients were younger than 25 years. Because prognostic parameters have not yet been reported, the authors present another case of a male age 14 years and a review of the available literature, which they conducted to determine prognostic parameters. METHODS: The medical literature about malignant mesothelioma of the tunica vaginalis testis was reviewed. For the determination of prognostic parameters, a univariate and multivariate Cox regression model was used to assess the relevance of the patient's age, history of asbestos exposure, tumor histology, primary therapeutic approach, and presence of metastatic disease to survival. RESULTS: Previous exposure to asbestos or asbestos-containing materials must be considered a risk factor for the development of malignant mesothelioma. The major difficulty in managing patients with malignant mesothelioma of the tunica vaginalis testis was determining an accurate preoperative diagnosis, which was reported in only two cases. Due to the lack of characteristic symptoms, 97.3% of the cases were diagnosed intraoperatively. Of patients who underwent local resection of the hydrocele wall, 35.7% experienced local tumor recurrence, as compared with 10.5% after scrotal orchiectomy and 11.5% after inguinal orchiectomy. Therefore, radical orchiectomy should be the first-line therapy. The median survival of the patients was 23 months, which decreased to 14 months in cases of recurrence. The overall recurrence rate (local and disseminated) was 52.5%. More than 60% of recurrences developed within the first 2 years of the follow-up. In some cases of disseminated mesothelioma, adjuvant chemotherapy or radiotherapy was given. Although reports on adjuvant treatments were limited, radiotherapy appeared to be more effective than chemotherapy. However, 37.9% died of disease progression. Assessment of prognostic parameters revealed a significant correlation of patient's age with survival (P < 0.01), with a better outcome for younger patients and a worse disease course for patients with primary disseminated disease (P < 0.05) in univariate analysis. A multivariate Cox regression model of prognostic parameters concerning survival did not yield statistically significant results. CONCLUSIONS: Malignant mesotheliomas of the tunica vaginalis testis rarely occur, but the possibility should be considered for all age groups. Univariate analysis determined that a patient's age and the presence of primary disseminated disease were prognostic parameters related to survival. Due to the invasive potential of this disease and the risk of tumor recurrence, radical orchiectomy and close follow-up are strongly recommended. PMID- 9874448 TI - Reduced expression of cyclin-dependent kinase inhibitor p27Kip1 is an indicator of malignant behavior in oral squamous cell carcinoma. AB - BACKGROUND: Reduced expression of the cyclin-dependent kinase inhibitor p27Kip1 has been reported to correlate with poor survival in cohorts of breast and colorectal carcinoma patients. Posttranslational ubiquitin-mediated proteasomal proteolysis is related to p27Kip1 protein levels. However, to the authors' knowledge, no previous study has examined the expression of p27Kip1 in oral squamous cell carcinoma (OSCC). METHODS: To examine the expression of p27Kip1 and its clinicopathologic roles in OSCC, the authors studied the expression of p27Kip1 protein by immunohistochemistry in deparaffinized tissue sections of 20 normal oral mucosa specimens, 22 epithelial dysplasia specimens, and 70 OSCCs, and analyzed its correlation with clinicopathologic parameters. They also studied the expression of p27Kip1 mRNA and protein in six OSCC cell lines by Northern blot and Western blot analysis. To examine the mechanism of reduced expression of p27Kip1, OSCC cell lines were treated with the proteasome inhibitor LLnV. RESULTS: All the normal oral mucosa specimens and 73% (16 of 22) of the oral epithelial dysplasia specimens expressed p27Kip1 at high levels, whereas 87% of the OSCCs (61 of 70) showed reduced expression of p27Kip1. Furthermore, the levels of expression of this protein were significantly lower in carcinomas with metastasis than those without metastasis. Although OSCC cell lines expressed p27Kip1 mRNA at various levels, most of them expressed p27Kip1 protein at lower or undetectable levels. LLnV induced the expression of p27Kip1 protein in HSC2 cells, in which p27Kip1 protein was originally undetectable. CONCLUSIONS: These findings suggest that 1) reduced expression of p27Kip1 may correlate with the development and progression of OSCC and can be an indicator of malignant behavior of this neoplasm, and 2) increased proteasome-mediated degradation may play an important role in the reduction of p27Kip1 protein expression. PMID- 9874449 TI - Nuclear accumulation of p53 in colorectal adenocarcinoma: prognostic importance differs with race and location of the tumor. AB - BACKGROUND: Although several studies have been conducted to examine the role of p53 genetic abnormalities and their prognostic value in colorectal carcinoma, the incidence of nuclear accumulation of p53 and the prognostic importance of nuclear accumulation of p53 in African-American and white patients have not been investigated separately. Therefore, the authors evaluated the prognostic significance of p53 nuclear accumulation in these two racial groups. METHODS: Nuclear accumulation of p53 was evaluated immunohistochemically in archival tissue specimens from 204 African-American and 300 white patients with primary colorectal adenocarcinomas who had undergone surgery. Survival times from colorectal adenocarcinoma were analyzed using Kaplan-Meier survival estimates and the Cox proportional hazards model for nuclear accumulation of p53 with adjustments for other confounding demographic and clinical variables. RESULTS: Approximately equivalent proportions of distal (54%) and proximal adenocarcinomas (47%) were positive for nuclear accumulation of p53 in African-American patients. In contrast, distal colorectal adenocarcinomas from white patients more frequently were positive for nuclear accumulation of p53 than adenocarcinomas of the proximal colon (63% vs. 38%, respectively). Nuclear accumulation of p53 was found to be a strong predictor of poor survival in white patients (hazard ratio = 6.77; P = 0.0001) but not in African-American patients with primary adenocarcinomas of the proximal colon. Nuclear accumulation of p53 was not of prognostic value in patients of either race with primary adenocarcinomas of the distal colorectum. CONCLUSIONS: Nuclear accumulation of p53 is a valuable indicator of poor prognosis only for white patients with adenocarcinomas of the proximal colon. The current study also suggests that the role of p53 dysregulation in colorectal adenocarcinomas may vary with the anatomic location of the tumor and the race of the patient. These findings suggest that the demographic characteristics of patients should be considered in the evaluation of prognostic markers of colorectal neoplasia. PMID- 9874450 TI - Serum alpha-L-fucosidase activity and early detection of hepatocellular carcinoma: a prospective study of patients with cirrhosis. AB - BACKGROUND: Serum alpha-L-fucosidase activity is considered a marker of hepatocellular carcinoma. To the authors' knowledge, its clinical usefulness in the early detection of hepatocellular carcinoma in the follow-up of cirrhotic patients has not been reported previously. METHODS: The authors prospectively studied serum alpha-L-fucosidase activity, in addition to alpha-fetoprotein and ultrasonography, in a regular screening of 132 cirrhotic patients during an 8 year follow-up. RESULTS: At enrollment, 120 patients had low alpha-L-fucosidase activity (below the cutoff value) and 12 had high activity. All patients had serum alpha-fetoprotein levels below the cutoff value. During the follow-up, hepatocellular carcinoma was detected in 19 patients, 16 with alpha-L-fucosidase activity below the cutoff value at enrollment and 3 with activity above it. In 7 of those 16 patients with carcinoma and low enzyme activity, the enzyme activity showed a significant increase 6-9 months before there was ultrasonographic evidence of a focal lesion, and by the time of diagnosis it had risen above the cutoff value in all of them; in only 3 of the 7 patients was the increase in alpha-L-fucosidase activity associated with an increase in alpha-fetoprotein. In another 4 of the 19 patients with carcinoma, only alpha-fetoprotein increased. CONCLUSIONS: Serum alpha-L-fucosidase activity is useful in the early detection of hepatocellular carcinoma. The data from this study suggest that cirrhotic patients who have a marked increase in serum alpha-L-fucosidase levels during follow-up should be closely monitored for signs of hepatocellular carcinoma development. PMID- 9874451 TI - A randomized Phase III trial of etoposide, epirubicin, and cisplatin versus 5 fluorouracil, epirubicin, and cisplatin in the treatment of patients with advanced gastric carcinoma. Turkish Oncology Group. AB - BACKGROUND: Gastric carcinoma is a substantial health problem in Turkey, and the majority of patients present with inoperable disease. The aim of this randomized trial was to assess the activity of 5-fluorouracil versus etoposide when combined with epirubicin plus cisplatin in patients with advanced gastric carcinoma (AGC). METHODS: In this prospective, randomized, multicenter Phase III study, previously untreated patients with histopathologically proven AGC enrolled after giving informed consent. Patients were allocated to receive either EEP (etoposide 120 mg/m2, epirubicin 30 mg/m2, and cisplatin 40 mg/m2 on Days 1 and 8) or FEP (5 fluorouracil 600 mg/m2, epirubicin 60 mg/m2, and cisplatin 80 mg/m2 on Day 1), and the regimens were to be repeated every fourth week. RESULTS: Of a total of 131 eligible patients, 64 were in the EEP group and 67 were in the FEP group. The objective response (complete + partial) rates for evaluable patients (n = 118) were comparable (P = 0.63) in the EEP (20.3%, 12/59) and FEP (15.3%, 9/59) groups, respectively. Actuarial analyses revealed comparable median progression free survival (6 vs. 7 months, P > 0.05) and overall survival (6 vs. 5 months, P > 0.05) duration in the EEP and FEP groups. Both regimens were well tolerated. The most common toxicity was Grade 1-2 nausea with or without vomiting. No chemotherapy-related death occurred. CONCLUSIONS: The current study resulted in inadequate response rates for EEP and FEP regimens. Neither combination, as used at the doses and schedules in this study, can be recommended as standard treatment for patients with AGC. PMID- 9874452 TI - Expression of nm23 in gastric carcinoma: association with tumor progression and poor prognosis. AB - BACKGROUND: Expression of nm23 has been shown to be inversely correlated with the metastatic potential of several human cancers. In the current study, the expression and prognostic impact of nm23 was immunohistochemically studied in 413 curatively resected gastric carcinomas. METHODS: Tumor sections of the 413 gastric carcinomas were stained with a polyclonal antibody that was raised against the nm23-H1/NDP kinase A, which is identical to the nm23-H1 gene product. RESULTS: Expression of nm23 was detected in 84.5% (n = 349) of all tumors, in the majority of cases (71.2%) causing a homogeneous staining reaction in more than 75% of tumor cells. Expression of nm23 was positively correlated with the intestinal type of tumor, according to the Lauren classification and advanced pT categories, and was also correlated with the presence of blood and lymphatic vessel invasion. In contrast, no correlation could be demonstrated between nm23 expression and lymph node involvement. As shown in univariate analysis, patients with nm23 positive tumors, especially those with nm23 positive diffuse-type carcinomas, had significantly shorter overall survival than patients with nm23 negative tumors (P = 0.03 and P = 0.0065, respectively). However, in a multivariate analysis that included the prognostic parameters pT category, pN category, and blood and lymphatic vessel invasion, this prognostic impact was not maintained. CONCLUSIONS: In contrast to results for breast and colorectal carcinomas, our results for 413 gastric carcinomas showed that expression of the designated metastasis suppressor gene nm23 is correlated with aggressive tumor growth and poor prognosis but is not an independent prognostic marker. PMID- 9874453 TI - Serum CA 125 level as a predictor of peritoneal dissemination in patients with gastric carcinoma. AB - BACKGROUND: Prediction of peritoneal dissemination is very difficult using current diagnostic tools such as computed tomography, ultrasonography, or various tumor markers. The predictive value of serum CA 125 levels for peritoneal metastasis from gastric carcinoma was studied. METHODS: The sera from 384 patients with gastric carcinoma were measured for CA 125 titer using an immunoradiometric assay. Carcinoembryonic antigen, carbohydrate antigen 19-9, and sialyl-Tn antigen were measured in the same samples. RESULTS: The serum CA 125 level was elevated according to the degree of peritoneal dissemination. The reference value for peritoneal dissemination was determined to be 35 U/mL, resulting in a sensitivity of 39.4%, specificity of 95.7%, and diagnostic accuracy of 90.8%. The diagnostic ability was more reliable than the other imaging modalities including computed tomography and ultrasonography and the other useful tumor markers for gastric carcinoma. The serum CA 125 level was elevated after gastrectomy for approximately 2 months, most likely due to the continuous inflammation of the peritoneum and lost predictive significance for peritoneal dissemination during this period. CONCLUSIONS: Measurement of the serum CA 125 titer may be a powerful predictor of peritoneal metastases in patients with gastric carcinoma. PMID- 9874454 TI - Tumor proliferation, p53 expression, and apoptosis in laryngeal carcinoma: relation to the results of radiotherapy. AB - BACKGROUND: Radiotherapy is used in the treatment of laryngeal carcinoma. The search for biologic parameters that could be used to identify patients who will respond to radiotherapy is crucial. The aim of this study was to determine whether the Ki-67 and p53 indices and the pretreatment apoptotic index would be useful in predicting local control and survival for a group of laryngeal carcinoma patients given postoperative radiotherapy. METHODS: Fifty-seven patients with laryngeal carcinoma treated between 1988 and 1993 were included in this study. Postoperative radiotherapy was given to a mean dose of 57.7 gray (Gy) (range, 50-68; median, 60) in 2-Gy daily fractions. Ki-67 and p53 immunostaining were performed on paraffin-embedded tissue. Cells were evaluated for apoptosis using hematoxylin and eosin-stained slides. Clinicopathologic tumor characteristics were studied in relation to Ki-67, p53, and apoptotic indices, and as prognostic factors for local control and survival in both univariate and multivariate analysis. RESULTS: The Ki-67, p53, and pretreatment apoptotic indices were not related to any clinicopathologic tumor characteristics. Five year actuarial local control for the whole group was 47%. Patients with tumors that had low Ki-67 proliferation had better long term local control (P < 0.01). and survival (P < 0.03). p53 expression was not predictive of local control or survival in this study. Patients with tumors that had low pretreatment apoptotic indices had better local control (P < 0.049) and survival (P < 0.056) than patients with highly apoptotic tumors. Tumor extension and the pretreatment apoptotic index were significant predictive factors for local control and survival in multivariate analysis. CONCLUSIONS: Ki-67 proliferation measurement and the pretreatment apoptotic index are useful in predicting the clinical outcome of laryngeal carcinoma patients referred for radiotherapy. The role of p53 oncoprotein determination in predicting these outcomes is unclear. Assessment of biologic tumor characteristics could aid in the selection of patients for different treatment strategies. PMID- 9874455 TI - Correlation of tumor size and axillary lymph node involvement with prognosis in patients with T1 breast carcinoma. AB - BACKGROUND: The prognosis of patients with T1 breast carcinoma remains controversial. Some studies have shown a low risk of lymph node metastasis and distant failure whereas others have not, possibly due to differences in the definition of tumor size. In this study, the authors assessed the relation between macroscopic tumor size, microscopic invasive tumor size, axillary lymph node involvement, and prognosis in a group of patients with clinically lymph node negative disease. METHODS: Between 1968 and 1986, 1865 women with American Joint Committee on Cancer clinical Stage I or II infiltrating carcinoma of the breast were treated at the Joint Center for Radiation Therapy with conservative surgery and radiation therapy. The study population was limited to 118 patients with clinically negative axillary lymph nodes for whom the macroscopic pathologic tumor size was identified unambiguously as being < or = 2.0 cm, who underwent an axillary lymph node dissection with at least 6 lymph nodes sampled, and for whom the microscopic size of the invasive component could be determined. The median follow-up time for surviving patients was 134 months (range, 90-208 months). No patients with pathologically negative axillary lymph nodes received systemic therapy. RESULTS: Macroscopic and microscopic tumor sizes differed by > 5 mm in 17 patients (14%), by 3-5 mm in 24 patients (20%), and by < or = 2 mm in 77 patients (65%). The macroscopic tumor size was smaller than the microscopic size in 37 patients (31%), larger in 55 patients (47%), and equal in 26 patients (22%). Pathologic axillary lymph node involvement was present in 21% of all patients. The risk of lymph node involvement was not significantly different for those patients with tumors < or = 1 cm compared with patients with tumors > or = 1.1 cm, regardless of whether tumor size was measured by macroscopic or microscopic examination. The 10-year actuarial rate of freedom from distant recurrence (FFDR) was 91% for lymph node negative patients with macroscopic tumors measuring < or = 1.0 cm compared with 77% for patients with macroscopic tumors measuring > or = 1.1 cm (P = 0.07). When measured microscopically, the rates were 96% and 72%, respectively (P = 0.001). CONCLUSIONS: There often is a discrepancy between microscopic tumor size and macroscopic tumor size. T1 tumors have a substantial risk of axillary lymph node metastasis whether measured macroscopically or microscopically. Among those patients with pathologic lymph node negative tumors who are not treated with systemic adjuvant therapy, microscopic invasive tumor size is a better predictor of 10-year FFDR than macroscopic tumor size. There is a substantial risk of distant failure for patients with tumors whose invasive component microscopically measure > or = 1.1 cm, whereas the prognosis for patients with tumors that microscopically measured < or = 1 cm is excellent. These results suggest that the microscopic size of the invasive component of breast carcinomas < or = 2.0 cm routinely should be reported. PMID- 9874456 TI - Breast carcinoma tumor characteristics in black and white women. AB - BACKGROUND: A significant disparity in mortality rates exists between black and white patients with breast carcinoma. This study was designed to compare breast carcinoma tumor characteristics by race and to examine the possible reasons for these differences. METHODS: Female patients with an initial diagnosis of breast carcinoma between January 1, 1985 and December 31, 1993 were selected from the Yale-New Haven Hospital Tumor Registry for this retrospective cohort study. All black patients were eligible and white patients were selected randomly and matched to each black patient by year of diagnosis. Data were gathered from multiple sources including the hospital, the Connecticut Tumor Registry, and the U. S. Census. All pathology specimens were reviewed at Yale-New Haven Hospital. RESULTS: The final cohort had 100 black and 300 white patients. The black patients tended to be younger than white patients at the time of diagnosis (mean age 55 years vs. 60 years; P = 0.001). A significant racial difference was noted in eight tumor characteristics: stage, size of the tumor, lymph node status, presence of necrosis, vascular/lymphatic invasion, ductal carcinoma in situ, perineural invasion, and progesterone receptor status. Although income, medical insurance coverage, and method of tumor detection explained some pathology differences, black patients still were more likely to have necrosis and a larger tumor size, even after adjustment. CONCLUSIONS: Black patients with breast carcinoma tend to be diagnosed at a younger age and in a few important respects have different tumor characteristics compared with white patients, even after controlling for income, medical insurance coverage, and method of tumor detection after screening mammography. These differences may have etiologic and clinical implications. PMID- 9874457 TI - Medical audit after 26,711 breast imaging studies: improved rate of detection of small breast carcinomas (classified as Tis or T1a,b) AB - BACKGROUND: Patients with breast carcinomas 1 cm or smaller are known to have the highest long term survival rates. This pathologic measurement was used as a quality indicator of the performance of different reporting radiologists. METHODS: A total of 26,711 diagnostic breast imaging studies were performed over a period of 5 years. Within the first 18 months, several general radiologists reported mammograms, whereas during the rest of the evaluated period only 1 radiologist was responsible for all mammography reporting. Radiologic findings leading to open biopsy were correlated with the histologic results. The cancer sizes from the histologic reports were recorded, and the number of Tis, T1a, and T1b cancers were correlated with all cancers found at open biopsy. The change in the relative percentage of small cancers was evaluated, along with the positive predictive value (PPV). RESULTS: Between 1991 and 1995, 26,711 breast imaging studies were performed. This resulted in 2698 open biopsies with 620 breast carcinomas detected. In the first 2 years, the percentages of small cancers detected (Tis or T1a,b) were 27.2% and 25.7%, respectively. In the third, fourth, and fifth years, the percentages increased to 38.8%, 34.5%, and 38.8%, respectively. At the same time, the PPV increased slightly, from 0.23 in 1991 to 0.27 in 1995. CONCLUSIONS: The detection of curable early stage breast carcinomas requires the dedication and commitment of a small group of radiologists who are willing to spend most of their time on this single subject. This will increase considerably the number of early stage cancers found and reduce the number of false-positive diagnoses. [See editorial counterpoint on pages 2430-1 and reply to counterpoint on pages 2432-3.] PMID- 9874458 TI - Preoperative CA 15-3 concentrations predict outcome of patients with breast carcinoma. AB - BACKGROUND: CA 15-3 is a breast-associated mucin that is elevated in the majority of breast carcinoma patients with distant metastases. Currently, the main application of this marker is in monitoring and detecting recurrences in patients with diagnosed breast carcinoma. METHODS: Preoperative serum concentrations (prior to excision of the primary tumor) of CA 15-3 were measured in 368 patients undergoing potentially curative surgical treatment for early breast carcinoma. These results were compared with prospectively recorded clinicopathologic characteristics and patient outcome data. RESULTS: A weak but significant positive association was found between CA 15-3 concentrations and both tumor stage and the number of involved axillary lymph nodes but not between CA 15-3 concentrations and estrogen receptor status. Patients with high concentrations of CA 15-3 had a significantly worse prognosis than patients with low concentrations. Using an optimum cutoff value of 30.38 U/mL, the probability of disease free survival at 5 years was 44% in patients with high CA 15-3 levels compared with 65% in patients with low CA 15-3 levels (P = 0.002, Mantel-Cox log rank test). The corresponding probabilities for overall survival were 67% and 83%, respectively (P < 0.001). The association of preoperative CA 15-3 levels with outcome was maintained in multivariate survival analysis and was not explained by the association between CA 15-3 and tumor size or lymph node burden. The relation between CA 15-3 and outcome also was found within some patient subgroups identified by traditional prognostic factors (axillary lymph node positive patients, patients with primary tumors >2 cm in greatest dimension, and patients with estrogen receptor positive tumors). CONCLUSIONS: Preoperative serum concentrations of CA 15-3 appear to have a significant relation to outcome in patients with early breast carcinoma and may have a role in the rational selection of patients for appropriate adjuvant treatments. To the authors' knowledge, CA 15-3 thus is one of the first circulating markers shown to be an independent prognostic indicator in patients with breast carcinoma. PMID- 9874459 TI - Biologic implications of the expression of vascular endothelial growth factor subtypes in ovarian carcinoma. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) has been identified as an important factor for tumor angiogenesis, which is essential for the growth, invasion, and metastasis of solid tumors. This study examines the clinical significance of VEGF subtypes in ovarian carcinoma. METHODS: Tumor specimens from 128 patients with ovarian carcinoma were evaluated for VEGF and its mRNA expression. The expression of VEGF, especially its subtypes, was determined by Western blot analysis with a sandwich enzyme immunoassay in ovarian carcinomas and reverse transcription-polymerase chain reaction and Southern blot analysis in normal ovaries that served as controls, and the relation between VEGF expression and the histologic types and clinical stages of ovarian carcinomas was analyzed. RESULTS: Among the four subtypes of VEGF, the populations of VEGF165 and VEGF121 were dominant in normal ovaries and ovarian carcinomas. The levels of VEGF and VEGF165 mRNA in ovarian carcinomas were significantly higher than in normal ovaries (P < 0.05). On the other hand, there was no significant difference in the levels of VEGF and VEGF165 mRNA among ovarian carcinomas classified according to histopathologic type or clinical stage. CONCLUSIONS: This analysis suggests that VEGF165 may be elevated in all stages of ovarian carcinoma via angiogenic activity, regardless of histopathologic type. PMID- 9874460 TI - Very frequent p53 mutations in metastatic prostate carcinoma and in matched primary tumors. AB - BACKGROUND: The frequency of mutant p53 in bone marrow metastases of patients with carcinoma of the prostate (CaP) and in matched sets of metastatic and primary lesions from the same patients was investigated. The data were examined in relation to prior treatment with androgen ablation (AA) therapy and were compared with the frequency of mutant p53 reported for primary CaP. METHODS: Seventeen patients with M1b (bone metastasis: TNM Stage IV) CaP had either unilateral or bilateral bone marrow biopsies taken for these studies. Specimens were divided and the outer one-third examined histologically to confirm the presence of CaP cells. Immunohistochemical (IHC) staining for accumulated p53 protein was performed by an antibody cocktail technique. RNA was extracted from the remaining portion of the biopsy, and p53 transcripts were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and screened for base sequence changes in the exon 4-11 region using nonisotopic single-strand conformation polymorphism (SSCP) analysis and direct DNA sequencing. RESULTS: Ten of 17 metastases (59%) demonstrated accumulation of p53. Six of 7 (86%) of the p53 IHC positive bone marrow samples contained RT-PCR-SSCP abnormalities, as did 2 of 3 IHC negative samples. Overall, 12 of 17 metastases (71%) contained mutant p53. Four of 7 biopsies (57%) retrieved prior to AA contained mutant p53, whereas 8 of 10 post-AA biopsies (80%) contained mutant p53. One patient showed identical SSCP abnormalities in right and left iliac crest metastases after therapy, and in this patient DNA sequencing demonstrated a missense mutation at codon 126 (TAC - > GGC, Tyr --> Gly). Archival primary cancers from seven patients were retrieved. All seven were IHC positive for p53 accumulation. CONCLUSIONS: p53 mutations are associated with increased metastatic potential of CaP. Abnormalities are found at approximately twice the frequency in metastases than in unselected samples of primary CaP, whereas in matched specimens there is a high rate of consonance. Mutant p53 may contribute to systemic therapy resistance, due to increased association with post-AA CaP specimens. PMID- 9874461 TI - The importance of human glandular kallikrein and its correlation with different prostate specific antigen serum forms in the detection of prostate carcinoma. AB - BACKGROUND: Human glandular kallikrein (hK2), the prostate specific antigen (PSA) close homologue, possesses approximately 80% structure identity with PSA. The identification of PSA was an important step in the detection of prostate carcinoma (PCa). Thus, hK2 measurement in the serum has the potential to become another important diagnostic test for PCa. In the current study, the authors measured the serum concentrations of the hK2 with "in-house" immunofluorometric assays in different patient groups. The correlation between serum hK2 and different PSA forms was investigated. METHODS: The prospectively collected serum samples were obtained preoperatively on admission from 311 consecutive male patients. Sixteen patients did not fulfill inclusion criteria; the remaining patients were divided into four groups (Groups I-III confirmed histologically): Group I: patients with PCa (n = 56); Group II: patients with benign prostatic hyperplasia (BPH) (n = 163); Group III: patients with BPH with a chronic in dwelling catheter (BPH cat) (n = 44); and Group IV-control group (n = 32). The patients in Group IV had urolithiasis, varicocele, or kidney or bladder tumors). An experimental immunofluorometric assay with an analytic sensitivity of 0.01 ng/mL and a functional sensitivity of 0.05 ng/mL was used to determine serum hK2 concentrations. Total PSA, free PSA, and PSA complexed to alpha-1 antichymotrypsin (PSA-ACT) also were measured. hK2 concentrations equal to or above the functional sensitivity limit were correlated with each of these PSA serum forms. Free to total PSA, hK2 to total PSA, and hK2 to free PSA ratios were calculated and compared in different patient groups. RESULTS: The hK2 concentrations were equal to or above the functional sensitivity limit in 179 of 311 samples (57.6%). In these samples, hK2 correlated best with free PSA (correlation coefficient [r] = 0.79) and correlated well with total PSA (r = 0.72) and PSA-ACT (r = 0.74). Similar correlations also could be observed when each clinical group was analyzed separately. The median proportion of hK2 in relation to total PSA was 2.1%, 1.8%, and 1.4%, respectively, for PCa, BPH, and BPH cat patients. Both the free to total PSA ratio and the hK2 to free PSA ratio discriminated well between PCa and BPH patients. Within the range of total PSA of 4-10 ng/mL (PCa [n = 11] and BPH [n = 41]) the hK2 to free PSA ratio had a specificity of 63.4% and 90.9% sensitivity (area under the receiver operating characteristic [ROC] curve = 0.85) whereas the free to total PSA ratio had a 34.1% specificity at the same sensitivity level (area under ROC curve = 0.74). CONCLUSIONS: The hK2 serum level correlates well with all PSA serum forms in all clearly defined clinical groups. The preliminary finding that the hK2 to free PSA ratio appeared to improve the detection of PCa compared with the free to total PSA ratio in patients with total PSA within a 4-10 ng/mL range is of clinical interest. Combining human serine proteases in the multivariate regression analysis will be a tool to improve cancer detection. Further investigations with more sensitive hK2 assays and in larger patient populations are needed to confirm this finding. PMID- 9874462 TI - Prognostic factors for survival in patients with brain metastases from renal cell carcinoma. AB - BACKGROUND: Patients presenting with brain metastases from renal cell carcinoma portend a poor prognosis, with a reported median survival of 4-6 months. Given their short life expectancy, these patients generally have been excluded from clinical trials that assess the efficacy of medical treatments. However, clinical impression suggests that some patients may achieve long term palliation. METHODS: The clinical features of 68 patients who were treated at the Institut Gustave Roussy for brain metastases from renal cell carcinoma were collected retrospectively. Using univariate and multivariate analyses, a prognostic model based on independent prognostic factors was established. An external data set of 57 patients was used to validate the model. RESULTS: The median survival was 7 months. On univariate analysis survival was related significantly to the following adverse prognostic factors: no initial nephrectomy, left side and temporal location of brain metastases, presence of fever or weight loss, erythrocyte sedimentation rate > 50 mm/h, and time from initial diagnosis to brain metastases < or = 18 months. Multivariate analyses identified the previous variable as well as the presence of other visceral metastases as independent prognostic factors. Forty-four patients (65%) with no or 1 adverse prognostic factor (average risk group) had a median survival of 8 months and a 26% 1-year survival rate. Twenty-four patients (35%) with 2 adverse prognostic factors (poor risk group) had a median survival of 3 months and a 1-year survival rate of 9%. This model proved to be discriminant in an external data set; the median survival of patients assigned to the average risk group was 11 months (46% 1-year survival rate) compared with 4 months (9% 1-year survival rate) for patients assigned to the poor risk group. CONCLUSIONS: Patients presenting with brain metastases from renal cell carcinoma and poor risk prognostic factors are highly unlikely to benefit from medical treatments except symptomatic procedures. Conversely, the enrollment of patients with average risk prognostic factors into clinical trials dealing with chemotherapy or immunotherapy may be considered. PMID- 9874463 TI - A combination of semiquantitative telomerase assay and in-cell telomerase activity measurement using exfoliated urothelial cells for the detection of urothelial neoplasia. AB - BACKGROUND: Telomerase is a ribonucleoprotein that synthesizes telomeres. It is detected in more than 85% of samples obtained from cancer tissues, including urologic neoplasia. The authors determined telomerase activity semiquantatively and in-cell telomerase activity in exfoliated urothelial cells obtained from urologic neoplasia specimens. The goal of this study was to provide additional information regarding a noninvasive approach to the detection of urologic neoplasia. METHODS: The authors used voided urine from 23 patients with urologic neoplasia, 2 patients with nonmalignant urologic disorders, and 10 normal individuals. Semiquantative determination of telomerase activity was performed using a fluorescence-based telomeric repeat amplification protocol (TRAP), and telomerase activity at the cellular level was determined by an in situ TRAP assay. RESULTS: The fluorescence-based TRAP assay detected urinary telomerase activity in samples from 10 of 13 patients with urologic neoplasia before treatment, whereas urinary cells obtained from 3 of 10 patients (including 1 patient with relapse) during or after treatment had detectable telomerase activity. In contrast, the in situ TRAP assay detected telomerase positive cells in samples from 11 of 13 patients before treatment and 6 of 10 patients during or after treatment. Of note was a dissociation of the results of the fluorescence based TRAP assay and those of the in situ TRAP assay for some patients. Some patients for whom telomerase activity was not detected with the fluorescence based TRAP assay had a low frequency of telomerase positive cells in their urine. CONCLUSIONS: A combination of semiquantative analysis and an in situ TRAP assay to detect telomerase positive cells might be a useful tool in the identification and monitoring of patients with urothelial neoplasia. PMID- 9874464 TI - Kappa-opioid receptor binding varies inversely with tumor grade in human gliomas. AB - BACKGROUND: Opioid agonists can inhibit cell proliferation in various neural tumor cell lines, including rat gliomas. Because opioid antimitogenic effects are mediated by opioid receptors, it was of interest to the authors to determine opioid receptor levels in human brain tumors. METHODS: Specimens obtained at craniotomy from 30 patients with glioma and nonneoplastic brain disorders were evaluated for their kappa-opioid receptor binding. Kd and Bmax values were estimated from homologous competition binding curves with the kappa1-selective radioligand [3H]U69,593. RESULTS: Receptor binding density was greatest in nonneoplastic brain tissue, less in Grade 2 and 3 astrocytoma, and least in glioblastoma multiforme. CONCLUSIONS: These results suggest that opioid receptor based stratification of grade may have clinical utility in distinguishing glioblastoma multiforme from lower grade astrocytomas, and thereby may facilitate diagnosis and treatment. PMID- 9874465 TI - Psychosocial and physical correlates of survival and recurrence in patients with head and neck carcinoma: results of a 6-year longitudinal study. AB - BACKGROUND: Studies that have examined correlations between psychosocial factors and survival in cancer patients do not permit any definitive conclusions. To the authors' knowledge, to date no study has examined the relation between medical as well as quality of life variables and survival in head and neck carcinoma patients. The current study focused on the complex interactions among psychosocial, medical, behavioral, and demographic variables as they relate to prognosis in these patients. METHODS: A total of 133 consecutive head and neck carcinoma patients were included in a prospective study at pretreatment. In addition to clinical variables, psychosocial and physical functioning was assessed by means of a self-report questionnaire. RESULTS: During the observation period 57 patients died whereas 76 were still alive at 6 years after treatment. Results of the multivariate survival analysis indicated that patients without head and neck metastasis had a better prognosis than patients with positive cervical lymph nodes. Pretreatment smoking showed a negative correlation with overall survival. Patients who were more physically self-efficacious (i.e., higher perceived physical abilities) were more likely to survive and less likely to develop a recurrence. In addition, patients who expressed intense psychosocial complaints prior to treatment had a better prognosis than had those who did not express such negative feelings. CONCLUSIONS: The current findings linking physical self-efficacy and prognosis are promising, but clinical trials are necessary to examine the direct and indirect mediational pathways of the variables that underlie physical efficacy and influence survival and recurrence. Also, the negative correlation between pretreatment smoking and survival suggests a need for increased efforts to address smoking in newly diagnosed patients. PMID- 9874466 TI - Immunotoxin combined with chemotherapy for patients with AIDS-related non Hodgkin's lymphoma. AB - BACKGROUND: The objective of this study was to develop and test a combined therapeutic approach for patients with AIDS-related lymphoma (ARL), employing agents with independent mechanisms of action and nonoverlapping toxicity. This study was designed to test the feasibility and tolerance of combining low dose chemotherapy with infusional immunotoxin in the treatment of ARL patients. METHODS: Previously untreated patients received low dose methotrexate, bleomycin, doxorubicin, cyclophosphamide, and vincristine (m-BACOD) on a 21- to 28-day schedule. Patients who did not have progressive disease by Cycle 3 received anti B4-blocked ricin (anti-B4bR), a murine monoclonal antibody linked to modified ricin, 20 microg/kg/day for 7 days administered by continuous infusion on an outpatient basis. A repeat cycle of anti-B4-bR was administered during Cycle 4 of chemotherapy based on tolerance. Patients received two cycles of chemotherapy beyond complete remission up to eight cycles. Study endpoints were toxicity, development of human antimurine antibody (HAMA) and human antiricin (HARA), tumor response, and survival. RESULTS: Twenty-six of 44 patients received the immunotoxin therapy. Anti-B4-bR infusion was associated with transaminase elevation (Grade 3) in 14 of 26 patients (58%), and flulike symptoms were common. HAMA or HARA was observed in 8 patients (31%). The overall response rate was 57% (13 complete responses and 12 partial responses). The median survival for all patients was 8.9 months. CONCLUSIONS: This study demonstrates the safety and feasibility of using chemotherapy and immunotoxin therapies in combination and supports their further evaluation to improve the outcomes of patients with ARL. PMID- 9874467 TI - What are cancer patients willing to pay for prophylactic epoetin alfa? A cost benefit analysis. AB - BACKGROUND: Anemia, one of the most common complications of cancer chemotherapy, has been managed with red blood cell (RBC) transfusions. As an alternative, the agent epoetin alfa has the potential to reduce the transfusion requirements of patients receiving cancer chemotherapy. To estimate the value that cancer patients place on the drug, an economic analysis using the concept of willingness to pay (WTP) was conducted. METHODS: The method of WTP was used within the framework of a classical cost-benefit analysis to estimate the net cost or benefit of administering prophylactic epoetin alfa to cancer patients. This estimate included the direct cost of epoetin alfa administration and savings secondary to reduced RBC transfusions. A cohort of 100 cancer patients who received or were scheduled to receive cisplatin or noncisplatin chemotherapy (50 per group) were then interviewed to measure the maximum WTP (net benefit) that they experienced with epoetin alfa. RESULTS: Regarding the benefits they would experience after 3 months of epoetin alfa administration, patients receiving cisplatin and noncisplatin therapy stated that they would be willing to pay an average of 587 U.S. dollars (U.S.$587) (95%CI: $300-$875) and U.S.$613 (95%CI: $324-$902), respectively. These benefits were then subtracted from the total cost of the drug when administered to patients receiving cisplatin (U.S.$3530) and noncisplatin (U.S.$3653) therapy. This produced a net incremental treatment cost of U.S.$2943 (95%CI: $2655-$3230) and U.S.$3039 (95%CI: $2750-$3328) for the respective treatment groups. CONCLUSIONS: The results of the current study suggest that the routine administration of epoetin alfa to cancer patients receiving myelosuppressive chemotherapy is a highly resource-intensive treatment policy with modest benefit to patients. Additional research is required to identify high risk patient subgroups who would benefit most from the drug. [See editorial on pages 2427-9, this issue.] PMID- 9874468 TI - Mississippi mud in the 1990s: risks and outcomes of vancomycin-associated toxicity in general oncology practice. AB - BACKGROUND: Discrepancies between the severity of toxicities reported in early clinical trials and recent clinical experience with vancomycin have led to confusion regarding the need for routine serum vancomycin level monitoring and discontinuation of vancomycin when toxicities occur. Therefore, the authors examined the incidence, outcomes, and predictive factors of vancomycin-associated toxicities in general oncology practice with the goal of developing clinically relevant prediction rules and guidelines. METHODS: All 742 consecutive cancer patients who received vancomycin at a comprehensive cancer center during a 3 month period were followed prospectively for the development and outcome of phlebitis, rash, ototoxicity, and nephrotoxicity. Logistic regression was used to derive a multiple variable model of the risk of nephrotoxicity. A clinical prediction rule, the Nephrotoxicity Risk Score, was developed from the risk model and validated prospectively. RESULTS: Phlebitis occurred in 3% of patients (95% confidence interval [95% CI], 2-4%), predominantly those with recently inserted central venous catheters. Rashes occurred in 11% of patients (95% CI, 9-13%); however, all but 4 patients also were receiving beta-lactam antibiotics. Clinical evidence of ototoxicity developed in 6% of patients (95% CI, 4-9%) who were receiving vancomycin plus other ototoxic agents and only 3% of patients (95% CI, 2-5%) not receiving other ototoxic agents (P = 0.08). Nephrotoxicity occurred in 17% of patients (95% CI, 15-20%). Logistic regression revealed that factors associated with an increased risk of nephrotoxicity included administration of other mild to moderate (P = 0.01) or severely nephrotoxic agents (P < 0.001) or an acute physiology and chronic health evaluation (APACHE) score > 40 (P = 0.002). Elevated serum vancomycin peak levels did not reliably predict subsequent nephrotoxicity. CONCLUSIONS: Vancomycin-associated toxicities usually are mild and self-limiting. Some patients are at a significantly higher risk of nephrotoxicity but the authors believe these individuals can be identified reliably with the Nephrotoxicity Risk Index using information available at vancomycin initiation. Further testing of the Nephrotoxicity Risk Index is ongoing. PMID- 9874469 TI - A longitudinal magnetic resonance imaging study of the brain in survivors in childhood acute lymphoblastic leukemia. AB - BACKGROUND: The objective of this study was to evaluate changes in magnetic resonance imaging (MRI) of the brain in children with acute lymphoblastic leukemia (ALL) during the first 5 years after the cessation of therapy and to correlate MRI abnormalities with neuropsychologic outcome. METHODS: Thirty-two children with ALL were studied at the end of treatment and 5 years later by brain MRI and the results were compared with the neuropsychologic findings. Fifteen patients had received chemotherapy alone and 17 had received chemotherapy plus cranial radiation. RESULTS: MRI of the brain was abnormal in 6 of 30 patients at the end of treatment and in 8 of 32 patients 5 years later. White matter changes (WMC) were found in 3 patients at the end of treatment and in 4 patients 5 years later. Two patients had developed new mild changes, whereas in one case WMC had normalized during the follow-up. Two patients had old hemorrhages or calcifications at each examination, with some improvement after follow-up, although one case revealed a new calcification or hemorrhage. Signs of cortical atrophy were observed in five patients at both evaluations. The patients with abnormal MRI findings did not differ significantly in their performance in the neuropsychologic tests from the patients with normal MRI findings, but the two patients with persistent WMC had a depression of verbal functions. CONCLUSIONS: Abnormalities in brain MRI were infrequent at the end of treatment for childhood ALL and 5 years later. They did not appear to correlate significantly with neuropsychologic outcome. Brain MRI is not very informative as a routine follow up method during the first 5 years after treatment. PMID- 9874470 TI - The National Cancer Data Base report on carcinoma of the gallbladder, 1989-1995. AB - BACKGROUND: Reports utilizing data from the Commission on Cancer's National Cancer Data Base (NCDB) have previously contained evaluations of time trends for stage of disease at diagnosis, treatment, and survival for multiple tumor sites. Data collected from 1989, 1990, 1994, and 1995 for carcinoma of the gallbladder are presented herein. METHODS: The data presented in this review were collected from hospital cancer registries from across the U.S. Seven calls for data yielded a total of 5,850,000 cases for the years 1985-1995, including 2574 gallbladder carcinoma cases from 1989-1990 and 2914 cases from 1994-1995 from hospital cancer registries across the U.S. These data represent approximately 8.8% and 8.4% of the estimated cases of liver and biliary track cancers diagnosed in the U.S. during the two respective time periods. RESULTS: There were no changes in patient demographics between 1989-1990 and 1994-1995. Most gallbladder carcinoma patients were white women. The documentation of tumor stage improved noticeably between the two study periods. There was no increased frequency in the occurrence of early stage tumors between the two reporting times, an era that saw the development and widespread application of laparoscopic cholecystectomy. Treatment differed according to stage of disease. Surgery alone, particularly nonradical surgery, was performed more frequently in the initial treatment of gallbladder carcinoma. A large percentage of gallbladder carcinoma patients received no definitive therapeutic intervention because of the advanced stage of disease at presentation and the lack of effective treatments for these cancers. Multimodality treatment was utilized more often for young patients. Survival was closely related to tumor stage, with 60% 5-year survival for Stage 0 patients, 39% for Stage I patients, and 15% for Stage II patients, but only 5% for Stage III patients and 1% for Stage IV patients. Patient outcome was not demonstrably affected by more aggressive therapy, nor was an adverse effect in results seen in early stage cancers between 1989-1990 and 1994-1995. CONCLUSIONS: The NCDB data are valuable in the evaluation of trends in malignant diseases, treatments, and patient survival. No substantial differences were apparent in the diagnosis, treatment, and survival of patients during this 7-year study period. The data do not support any adverse effect on outcome results with the introduction of laparoscopic cholecystectomy in the U.S. PMID- 9874471 TI - The National Cancer Data Base report on endometrial carcinoma in African-American women. AB - BACKGROUND: Although the incidence of uterine carcinoma is lower among African American women compared with white women, the mortality rates are higher for African-American patients. This report is part of an ongoing series on gynecologic malignancies in African-American women. METHODS: Hospital registry reports collected by the National Cancer Data Base were used to describe some of the differences in case presentation and management characteristics of endometrial carcinoma in these two groups. The cases represented 52,307 Non Hispanic white and 3226 African-American women diagnosed with primary carcinoma of the endometrium between 1988-1994. RESULTS: More African-American patients were diagnosed with less favorable histologies than white patients, at more advanced stages of disease, and with less tumor differentiation. Income had no effect on stage or grade. African-American patients were treated less often for their tumor at every stage of diagnosis compared with white women. Income generally had no effect on whether treatment was provided, but limited income was associated with a lack of treatment in African-American patients with American Joint Committee on Cancer Stage IV tumors. African-American women were less frequently treated surgically and, among surgically treated patients at advanced stages of disease, they received adjuvant radiotherapy less often and chemotherapy more often than white patients. Five-year survival was poorer for African-American women, even for patients with the more favorable Stage I adenocarcinoma who were treated surgically. CONCLUSIONS: All patients, regardless of race, should be treated appropriately as dictated by medical and prognostic factors and not by race. Although no screening methods currently exist for endometrial carcinoma, the development of procedures for identifying patients at risk for the prognostic factors that lead to a poor outcome should be a primary focus. PMID- 9874472 TI - A National Cancer Data Base report on 53,856 cases of thyroid carcinoma treated in the U.S., 1985-1995 [see commetns]. AB - BACKGROUND: The National Cancer Data Base (NCDB) represents a national electronic registry system now capturing nearly 60% of incident cancers in the U. S. In combination with other Commission on Cancer programs, the NCDB offers a working example of voluntary, accurate, cost-effective "outcomes management" on a both a local and national scale. In addition, it is of particular value in capturing clinical information concerning rare cancers, such as those of the thyroid. METHODS: For the accession years 1985-1995, NCDB captured demographic, patterns of-care, stage, treatment, and outcome information for a convenience sample of 53,856 thyroid carcinoma cases (1% of total NCDB cases). This article focuses on overall 10-year relative survival and American Joint Committee on Cancer (AJCC) (3rd/4th edition) stage-stratified 5-year relative survival for each histologic type of thyroid carcinoma. Care patterns also are discussed. RESULTS: The 10-year overall relative survival rates for U. S. patients with papillary, follicular, Hurthle cell, medullary, and undifferentiated/anaplastic carcinoma was 93%, 85%, 76%, 75%, and 14%, respectively. For papillary and follicular neoplasms, current AJCC staging failed to discriminate between patients with Stage I and II disease at 5 years. Total thyroidectomy +/- lymph node sampling/dissection represented the dominant method of surgical treatment rendered to patients with papillary and follicular neoplasms. Approximately 38% of such patients receive adjuvant iodine 131 ablation/therapy. At 5 years, variation in surgical treatment (i.e., lobectomy vs. more extensive surgery) failed to translate into compelling differences in survival for any subgroup with papillary or follicular carcinoma, but longer follow-up is required to evaluate this. NCDB data appeared to validate the AMES prognostic system, as applied to papillary cases. Younger age appeared to influence prognosis favorably for all thyroid neoplasms, including medullary and undifferentiated/anaplastic carcinoma. NCDB data also revealed that unusual patients diagnosed with undifferentiated/anaplastic carcinoma before age of 45 years have better survival. CONCLUSIONS: The NCDB system permits analysis of care patterns and survival for large numbers of contemporaneous U. S. patients with relatively rare neoplasms, such as thyroid carcinoma. In this context, it represents an unsurpassed clinical tool for analyzing care, evaluating prognostic models, generating new hypotheses, and overcoming the volume-related drawbacks inherent in the study of such neoplasms. [See editorial on pages 2434-6, this issue.] PMID- 9874473 TI - Regional diversity and breadth of the National Cancer Data Base. AB - BACKGROUND: The National Cancer Data Base (NCDB), a joint project of the Commission on Cancer of the American College of Surgeons and the American Cancer Society, is a cancer management and outcomes data base for health care organizations. It provides a comparative summary of patient care that is used by participating hospitals and communities for self-assessment. This article describes the most current (1995) data. METHODS: Since 1989, 7 calls for data have been issued, yielding a total of 5,558,389 cancer patient reports for the years 1985-1995. A total of 1849 hospital cancer registries have participated in at least 1 of the calls for data. RESULTS: One thousand one hundred and fourteen hospitals from 50 states and the District of Columbia reported 655,627 cases for the diagnosis year 1995. The hospitals represented a wide range of sizes (187 [16.8%] with 1000+ cases annually, 405 [36.4%] with 500-999 cases annually, 255 [22.9%] with 300-499 cases annually, 211 [18.9%] with 100-299 cases annually, and 56 [5%] with < 100 cases annually) and types (21 [1.9%] National Cancer Institute [NCI]-recognized cancer centers, 119 [10.7%] government hospitals, 102 [9.2%] teaching hospitals, 256 [23.0%] large community hospitals, 297 [26.7%] medium/small community hospitals, and 257 [23.1%] nongovernmental hospitals without approval status from the Commission on Cancer or NCI recognition). Remarkably similar distributions of cases by primary site and age were reported from each of six U.S. geographic regions. In addition, within each of these six regions, the cases were reported from a wide range of income strata and ethnicities. For several states, relatively few cancer cases were reported. For several examples of relatively rare patient and tumor groups, all reported cases between 1985-1995 included potentially useful quantities of patients in whom further study of such special groups was warranted. CONCLUSIONS: The authors conclude that the reported cases most likely are representative at the regional (but not state) level of cancer patients diagnosed and treated at U.S. hospitals with regard to types of cancer and ages of the patients. They conclude further that cancer reporting may be quite diverse within each region with regard to other known patient and reporting institution characteristics. PMID- 9874474 TI - Reluctant remyelination: the missing precursors. PMID- 9874475 TI - Oligodendrocyte progenitors are present in the normal adult human CNS and in the lesions of multiple sclerosis. AB - In multiple sclerosis, partial remyelination is conspicuous in many lesions, but widespread and lasting myelin repair ultimately fails as disability and handicap accumulate. Thus far, the precise identity of the cell responsible for limited spontaneous myelin repair has remained obscure. In the rodent, the proliferative oligodendrocyte progenitor is the most efficient remyelinating cell; this has now been identified in cultures prepared from normal human brain, but has proved difficult to demonstrate in situ. We adapted techniques using antibodies against the human platelet-derived growth factor-alpha receptor to identify oligodendrocyte progenitors in human tissue sections. Small numbers of oligodendrocyte progenitors were found in normal adult human white matter. Progenitors were also demonstrable in acute and chronic lesions from patients dying with multiple sclerosis, but with no evidence of any marked reactive increase in cell numbers. Understanding the biology of the remyelinating cell, and in particular the reason for its apparent failure to repopulate demyelinated lesions, is important for the development of remyelination treatments. PMID- 9874476 TI - Perisylvian dysgenesis. Clinical, EEG, MRI and glucose metabolism features in 10 patients. AB - We studied 10 patients who had neurological disorders with a MRI-based diagnosis of perisylvian dysgenesis based on the fact that the parasagittal and centrifugal extremity of the sylvian fissure was abnormally mesial. This abnormality was bilateral in seven cases; in the other three patients, the contralateral sylvian fissure appeared either normal (two cases) or enlarged (open operculum). The perisylvian cortex had a polymicrogyric appearance in most patients. Potential aetiopathogenic factors were determined in four patients. In two of them, ischaemia at mid-gestation was ascribed to death of a co-twin in a context of monozygotic twinning. In the other two patients, who were siblings, genetic factors were suspected. Pseudobulbar palsy was found in eight patients and epilepsy in five patients. We used PET with [18F]fluorodeoxyglucose to test the hypothesis that, despite this clinical and MRI heterogeneity, regional cerebral glucose distribution could have common features in these patients. The analysis of PET data was performed by visual inspection in two cases and by using statistical parametric mapping (SPM) in eight patients compared with a control group. Segmented grey matter MRIs of seven out these patients were also analysed using SPM. We found that the abnormal perisylvian cortex had normal grey matter activity in eight patients and in the other two there was a heterogeneous pattern with areas of preserved metabolism and of decreased metabolism. Metabolic changes were also detected outside the polymicrogyric-like cortex; three patients had hypometabolic areas in cortical regions where the MRI appeared normal and had a normal intensity. When polymicrogyria extended into the white matter, this ectopic dysgenetic cortex was associated with a grey matter pattern within the white matter territory, and was detected by SPM as areas of PET hypermetabolism and MRI hyperintensity. In order to detect possible metabolic changes undetected by the individual analyses, the group of patients was compared with the control group. This comparison revealed bilateral hypometabolism in the frontal opercular cortex. We propose that these PET data be considered in light of the presumed cyto-architectonic pattern of perisylvian dysgenesis, i.e. polymicrogyria. In this malformation, two dense cell layers are separated by a necrotic sparse cell layer. We speculate that the amount of synaptic activity preserved in these dense cell layers depends on the importance and timing of the necrotic process; this hypothesis accounts for the large range of metabolic patterns found, from profoundly decreased glucose metabolism to nearly normal activity. PMID- 9874477 TI - Correlation of quantitative tests of nerve and target organ dysfunction with skin immunohistology in leprosy. AB - Loss of nociception and hypohidrosis in skin are hallmarks of leprosy, attributed to early invasion by Mycobacterium leprae of Schwann cells related to unmyelinated nerve fibres. We have studied skin lesions and contralateral clinically unaffected skin in 28 patients across the leprosy spectrum with a range of selective quantitative sensory and autonomic tests, prior to biopsy of both sites. Unaffected sites showed normal skin innervation, when antibodies to the pan-neuronal marker PGP (protein gene product) 9.5 were used, with the exception of intraepidermal fibres which were not detected in the majority of cases. Elevation of thermal thresholds and reduced sensory axon-reflex flare responses in affected skin correlated with decreased nerve fibres in the subepidermis, e.g. axon-reflex flux units (means+/-SEM) for no detectable innervation; decreased innervation; and clinically unaffected skin, were 23+/ 3.1; 41.2+/-7.3; and 84.5+/-4.0, respectively. Reduced nicotine-induced axon reflex sweating was correlated with decreased innervation of sweat glands. Where methacholine-induced direct activation of sweat glands was affected, there was inflammatory infiltrate and loss of sweat gland structure. This study demonstrates a correlation between selective nerve dysfunction on clinical tests and morphological changes in skin, irrespective of the type of leprosy, and is the first to show that loss of sweating in leprosy may result either from decreased innervation and/or involvement of the sweat glands. The findings have implications for the selection and monitoring of patients with leprosy in clinical trials which aim to restore cutaneous function. PMID- 9874478 TI - Trajectories of cholinergic pathways within the cerebral hemispheres of the human brain. AB - All sectors of the human cerebral cortex receive dense cholinergic input. The origin of this projection is located in the Ch4 cell group of the nucleus basalis of Meynert. However, very little is known about the location of the pathways which link the cholinergic neurons of the nucleus basalis to the human cerebral cortex. This question was addressed in whole-hemisphere sections processed for the visualization of multiple cholinergic markers. Two highly organized and discrete bundles of cholinergic fibres extended from the nucleus basalis to the cerebral cortex and amygdala and were designated as the medial and lateral cholinergic pathways. These bundles contained acetylcholinesterase, choline acetyltransferase and nerve growth factor receptors, confirming their cholinergic nature and origin within the basal forebrain. The medial pathway joined the white matter of the gyrus rectus, curved around the rostrum of the corpus callosum to enter the cingulum and merged with fibres of the lateral pathway within the occipital lobe. It supplied the parolfactory, cingulate, pericingulate and retrosplenial cortices. The lateral pathway was subdivided into a capsular division travelling in the white matter of the external capsule and uncinate fasciculus and a perisylvian division travelling within the claustrum. Branches of the perisylvian division supplied the frontoparietal operculum, insula and superior temporal gyrus. Branches of the capsular division innervated the remaining parts of the frontal, parietal and temporal neocortex. Representation of these cholinergic pathways within a 3D MRI volume helped to identify white matter lesion sites that could interfere with the corticopetal flow of cholinergic pathways. PMID- 9874479 TI - Perception of movement and shape in Alzheimer's disease. AB - Alzheimer's disease is the most prevalent cause of abnormal cognitive decline in older adults and commonly affects visual function. Recent evidence suggests that Alzheimer's disease impairs the processing of visual motion, but these conclusions are based on conflicting results in the few cases studied, and the processing of complex motion images has not been investigated. In the present study of motion processing in Alzheimer's disease we assessed visual functions in 63 adults: 41 with mild to moderate Alzheimer's disease (mean age 72.3 years) and 22 without dementia (mean age 71.7 years). Processing of motion cues was tested with computer animation sequences known as random-dot cinematograms, which resemble the stimuli used to define motion processing deficits in primates with lesions of cortical area MT. Results showed that participants with Alzheimer's disease required significantly greater thresholds for perceiving shapes defined by motion cues compared with participants without dementia (P = 0.0005). There were no significant differences between the two groups (P < 0.05) in static visual acuity, static spatial contrast sensitivity and, surprisingly, dynamic visual acuity, which was normal, and motion direction discrimination, which was relatively spared. We conclude that mild to moderate Alzheimer's disease has significant effects on the perception of structure from motion with relative sparing of motion direction discrimination. We cannot exclude a contribution by retinal pathology, but retinal dysfunction alone cannot explain the pattern of defects we observed. The complex motion image processing deficit we identified is likely to have a cerebral basis and has the potential to affect navigation and the recognition of objects in relative motion, as encountered during walking and automobile driving. PMID- 9874480 TI - Functional mapping of human sensorimotor cortex with electrocorticographic spectral analysis. I. Alpha and beta event-related desynchronization. AB - Human scalp EEG studies have shown that event-related desynchronization (ERD) in the alpha (8-13 Hz) and beta (15-25 Hz) bands may be used to detect functional activation of sensorimotor cortex. However, in most previous studies somatotopy has not been examined in detail and brief, self-paced movements, focusing on the planning of motor output, have been used. We recorded electrocorticographic (ECoG) signals in five clinical subjects during a visual-motor decision task that was designed to activate the representations of different body parts in sensorimotor cortex. To focus more on execution of motor output than on its planning, subjects were instructed to make sustained isometric muscle contractions in different body parts (tongue protrusion, fist-clenching or foot dorsiflexion) in response to randomized visual stimuli depicting each action. ECoG spectral analysis utilized a mixed-effects analysis of variance model in which within-trial temporal dependencies were taken into account, and the magnitude and statistical significance of alpha and beta ERDs were mapped onto a surface rendering of each subject's brain MRI. Cortical electrical stimulation was performed in all subjects for clinical purposes, and the resulting maps of sensorimotor function were compared with those generated by ECoG spectral analysis. During the early phases of the motor responses, alpha ERD commonly occurred in a diffuse spatial pattern that was not somatotopically specific. During the late phases, the spatial pattern of alpha ERD usually became more focused and somatotopically specific. Maps of alpha ERD were closer to cortical stimulation maps when alpha ERD was sustained throughout the late phases of the motor responses. Thus, the topography of alpha ERD more resembled traditional somatotopy when its temporal profile approximated that of the motor response. The topography of beta ERD was often more discrete and somatotopically specific than that of alpha ERD, but beta ERD was often transient and sometimes absent. Sometimes, unilateral limb movement produced sustained alpha and beta ERD over bilateral sensorimotor cortices, with overlapping patterns for different body parts. The topographical spread of alpha ERD beyond expected functional anatomical boundaries during early (and sometimes late) phases of motor responses invites a re-examination of traditional assumptions about sensorimotor functional neuroanatomy, as well as the role of alpha ERD as an index of cortical activation. We agree with others that the somatotopic representations of different body parts overlap more than previously thought. Also, unilateral limb movements may be associated with both contralateral and ipsilateral activation of sensorimotor cortex. We conjecture that alpha ERD may reflect activity within a broad synaptic network with distributed cortical representations. PMID- 9874481 TI - Functional mapping of human sensorimotor cortex with electrocorticographic spectral analysis. II. Event-related synchronization in the gamma band. AB - It has been shown in animals that neuronal activity in the 'gamma band' (>30 Hz) is associated with cortical activation and may play a role in multi-regional and multi-modal integration of cortical processing. Studies of gamma activity in human scalp EEG have typically focused on event-related synchronization (ERS) in the 40 Hz band. To assess further the gamma band ERS further, as an index of cortical activation and as a tool for human functional brain mapping, we recorded subdural electrocorticographic (ECoG) signals in five clinical subjects while they performed visual-motor decision tasks designed to activate the representations of different body parts in sensorimotor cortex. ECoG spectral analysis utilized a mixed-effects analysis of variance model in which within trial temporal dependencies were accounted for. Taking an exploratory approach, we studied gamma ERS in 10-Hz-wide bands (overlapping by 5 Hz) ranging from 30 to 100 Hz, and compared these findings with changes in the alpha (8-13 Hz) and beta (15-25 Hz) bands. Gamma ERS (observed in three out of subjects) occurred in two broad bands-'low gamma' included the 35-45 and 40-50 Hz bands, and 'high gamma' the 75-85, 80-90, 85-95 and 90-100 Hz bands. The temporal and spatial characteristics of low and high gamma ERS were distinct, suggesting relatively independent neurophysiological mechanisms. Low gamma ERS often began after onset of the motor response and was sustained through much of it, in parallel with event-related desynchronization (ERD) in the alpha band. High gamma ERS often began during, or slightly before, the motor response and was transient, ending well before completion of the motor response. These temporal differences in low and high gamma suggest different functional associations with motor performance. Compared with alpha and beta ERD, the topographical patterns of low and high gamma ERS were more discrete and somatotopically specific and only occurred over contralateral sensorimotor cortex during unilateral limb movements (alpha and beta ERD were also observed ipsilaterally). Maps of sensorimotor function inferred from gamma ERS were consistent with maps generated by cortical electrical stimulation for clinical purposes. In addition, different task conditions in one subject produced consistent differences in both motor response latencies and onset latency of gamma ERS, particularly high gamma ERS. Compared with alpha and beta ERD, the topography of gamma ERS is more consistent with traditional maps of sensorimotor functional anatomy. In addition, gamma ERS may provide complementary information about cortical neurophysiology that is useful for mapping brain function in humans. PMID- 9874482 TI - Visual peripersonal space centred on the face in humans. AB - A convergent series of studies in monkeys and man suggests that the computation of visual space is performed in several brain regions for different behavioural purposes. Among these multiple spatial areas, the ventral intraparietal cortex, the putamen and the ventral aspect of the premotor cortex (area 6) contain a system for representing visual space near the face (peripersonal space). In these cerebral areas some neurons are bimodal: they have tactile receptive fields on the face, and they can also be driven by visual stimuli located near the tactile field. The spatial correspondence between the visual and tactile receptive fields provides a map of near visual space coded in body-part-centred co-ordinates. In the present study we demonstrate for the first time the existence of a visual peripersonal space centred on the face in humans. In patients with right hemispheric lesions, visual stimuli delivered in the space near the ipsilesional side of the face extinguished tactile stimuli on the contralesional side (cross modal visuotactile extinction) to the same extent as did an ipsilesional tactile stimulation (unimodal tactile extinction). Furthermore, a visual stimulus presented in the proximity of the contralesional side of the face improved the detection of a left tactile stimulus: i.e. under bilateral tactile presentation patients were more accurate to report the presence of a left tactile stimulus when a simultaneous visual stimulus was presented near the left side of the face. However, when visual stimuli were delivered far from the face, visuotactile extinction and visuotactile facilitation effects were dramatically reduced. These findings are consistent with the hypothesis of a representation of visual peripersonal space coded in bodypart-centred co-ordinates, and they provide a striking demonstration of the modularity of human visual space. PMID- 9874483 TI - Matrix metalloproteinase-9 (gelatinase B) is selectively elevated in CSF during relapses and stable phases of multiple sclerosis. AB - Matrix metalloproteinases (MMPs) are a family of endopeptidases capable of enzymatic digestion of subendothelial basement membrane and other components of the extracellular matrix. Expression of MMP-2, -3, -7 and -9 is increased around multiple sclerosis plaques and in brain tissue in experimental allergic encephalomyelitis. To measure quantitatively the expression of these MMPs and their endogenous inhibitors (TIMP-1 and -2), we analysed samples from 52 patients with relapsing-remitting and primary progressive multiple sclerosis by ELISA (enzyme-linked immunosorbent assay) and substrate-gel electrophoresis (zymography). MMP-9 was increased over controls in 100% of relapsing-remitting multiple sclerosis cases, with similar levels detected in relapses and clinically stable phases of disease. In primary progressive multiple sclerosis, MMP-9 was increased in 57% of CSF samples, but concentrations were below those encountered in the relapsing-remitting form. The selective upregulation of MMP-9 suggests that T-cells and macrophages invading the brain parenchyma and the CSF space are the predominant source of MMP-9 in multiple sclerosis. TIMPs and other MMPs (MMP 2 and -3) were not upregulated or not detectable (MMP-7) in CSF of patients with relapsing-remitting and primary progressive multiple sclerosis. The sustained increase of MMP-9 in clinically stable multiple sclerosis supports the concept that multiple sclerosis is associated with ongoing proteolysis that may result in progressive tissue damage. The selective inhibition of MMP-9 could be a useful approach for the prevention of disease progression in multiple sclerosis. PMID- 9874484 TI - The role of DYT1 in primary torsion dystonia in Europe. AB - Primary torsion dystonia (PTD) is a clinically and genetically heterogeneous movement disorder. DYT1 on chromosome 9q34 was the first PTD gene to be mapped. A 3-bp (GAG) deletion in this gene was reported to account for almost all early limb-onset generalized PTD. No relationship has been found with DYT1 in patients with prominent craniocervical involvement. To elucidate the DYT1-associated phenotype, we analysed the DYT1 mutation in 150 PTD patients, either sporadic or index cases from small PTD families. Twenty-two patients were positive for the GAG deletion in the DYT1 gene. Fifteen of them presented with the typical DYT1 phenotype (early, limb-onset generalized dystonia without spread to craniocervical muscles), four had limb-onset dystonia with spread to craniocervical muscles, two patients had arm-onset segmental dystonia and one had focal right-arm dystonia. One-hundred and twenty-eight patients were negative for the DYT1 mutation. Forty-six of them had segmental dystonia and 59 had focal dystonia. The other 23 patients presented with generalized dystonia, either with craniocervical involvement (13 patients) or without spread to the craniocervical region (typical DYT1 phenotype-10 patients). These data confirm the importance of the GAG deletion in European cases of PTD, and indicate phenotypic and genotypic heterogeneity. PMID- 9874485 TI - A clinicogenetic analysis of six Indian spinocerebellar ataxia (SCA2) pedigrees. The significance of slow saccades in diagnosis. AB - Clinical revaluation and genetic analysis of six Indian pedigrees, segregating autosomal dominant cerebellar ataxia, slow saccades and peripheral neuropathy, has been undertaken, and expansion at the spinocerebellar ataxia 2 (SCA2) locus was confirmed in 14 affected family members. These families became available from 31 phenotypically similar families seen over the years. In common with other neurodegenerative disorders resulting from expansion of a CAG trinucleotide repeat motif, an inverse correlation between repeat size and age at onset and severity is observed, although the size range (36-45 repeat units) for the expanded alleles is comparatively limited. Saccadic velocity was reduced in all our patients, even in the early stages of the disease. The observation of slow saccades in affected individuals has been proposed previously as an important diagnostic criterion serving to distinguish the SCA2 phenotype. This is now confirmed in a retrospective study of the clinical literature, facilitated by the cloning of the SCA2 gene and the subsequent genetic analysis of families segregating this phenotype. We therefore argue that the clinical appraisal of 'ophthalmoplegia' be subject to more precise definition, as differentiation between the various types of ocular dysfunction can be an important adjunct to diagnosis. PMID- 9874486 TI - Space exploration in neglect. AB - The present study investigated the gaze as well as the head and the eye-in-head movements of neglect patients while they were exploring their surroundings. A random configuration of letters was presented on the inner surface of a sphere that surrounded the subject, requiring free exploratory eye and head movements. The subjects were requested to search for a single (non-existent) target letter. The co-ordination of eye and head movements in patients with neglect resembled the pattern usually observed in healthy subjects orienting to eccentric visual targets. They performed hypometric head movements with additional shifts of eye in-head position. Moreover, like healthy subjects, the patients with neglect explored space with gaze, with head and eye-in-head movements that were symmetrically distributed around preferred orientations in space. However, in contrast to controls, these centres of exploration were shifted towards the right. The average horizontal position of gaze and of head movements lay right of the body's mid-sagittal plane, the average eye-in-head position right of the head midline. The preferred orientations were located far away from the anatomical limits of horizontal gaze, head and eye-in-head movements. The decrease of exploration towards more eccentric locations left and right of these orientations thus could not be explained by anatomical restrictions. The results argue against a model of neglect that proposes a lateral gradient of attentional orienting towards the ipsilesional side. Exploring the surroundings, the patients did not orient gaze, the head or the eyes in the head towards the extreme ipsilesional side, nor even close to it. The results favour a deviation model suggesting a shift of the whole frame for exploratory behaviour towards the ipsilesional side. In addition to this shift, we found a second component of altered visual exploration in neglect. The patients' head and gaze movements exhibited a reduced variability around the deviated centre of exploration. The variability was not generally reduced but rather concerned specifically the horizontal dimension. The latter was found even when the area of exploration was paralleled between the groups, requiring the control subjects to search only in that part of the letter array that the neglect patients had explored spontaneously. Possible mechanisms, such as a disturbed ability to update the spatial representation of visual targets or an altered neural representation of space in the horizontal dimension, are discussed. PMID- 9874487 TI - Morphology of the planum temporale and corpus callosum in left handers with evidence of left and right hemisphere speech representation. AB - In the present study we investigated planum temporale asymmetry and corpus callosum morphology in a sample of young adult left-handed males, using MRI. Two subgroups of left-handed males were identified on the basis of their differing speech lateralization patterns, which were inferred from results of the Fused Dichotic Words Test. These individuals then underwent MRI in order to obtain area measurements of the left and right planum temporale and the midsagittal corpus callosum. Comparisons between these left-handed males and an archival sample of age-matched right-handed males were also performed. Results demonstrated a strong leftward asymmetry in the planum temporale among subjects with left-hemisphere speech representation, regardless of handedness, but no consistent planum temporale asymmetry among subjects with right hemisphere speech representation. The results suggest that reversed speech lateralization is not necessarily accompanied by a concomitant reversal of planum temporale asymmetry. Examination of callosal areas revealed that left-handed subjects with left hemisphere speech functions had a larger corpus callosum than either left-handed subjects with right hemisphere speech functions or right-handed subjects. Increased interhemispheric communication may be required when the neural systems underlying speech and handedness are represented in opposite hemispheres. PMID- 9874488 TI - Abnormal development of biceps brachii phasic stretch reflex and persistence of short latency heteronymous reflexes from biceps to triceps brachii in spastic cerebral palsy. AB - Co-contraction of antagonist muscles is characteristic of spasticity arising from perinatal brain damage but not in spasticity occurring after brain damage in adulthood. Such co-contraction is a normal feature of early post-natal motor development. Heteronymous, monosynaptic Group Ia projections from biceps brachii to both the antagonist triceps brachii and to other synergist and non-synergist muscles of the upper limb occur in the newborn baby and become restricted during the first 4 years to motor neurons of primarily synergistic muscles. Longitudinal and cross-sectional studies have been performed to test the hypothesis that inappropriate heteronymous excitatory projections persist in children with perinatal brain damage who develop spasticity. Subjects with spasticity, from brain damage acquired in adulthood were also studied to determine if these projections simply become unmasked as part of spasticity, independent of the age of occurrence of the brain damage. Twenty-nine healthy newborn babies and 29 at high risk for cerebral palsy, 12 of whom developed spastic quadriparesis, were studied longitudinally for 4 years. Thirty-eight subjects, aged 8-30 years, with spasticity of perinatal origin (11 hemiplegic, 11 quadriplegic, 16 with Rett syndrome) and 11 subjects with stroke in adulthood and spastic hemiplegia were also studied. The results were compared with those obtained in 372 normal subjects aged from birth to 55 years. Small taps were delivered to the tendon of biceps brachii using an electromechanical tapper. Surface EMG was recorded from biceps and triceps brachii, pectoralis major and deltoid. In the longitudinal study, those developing spastic quadriparesis showed persistent low thresholds for the homonymous phasic stretch reflex, which had abnormally short onset latencies. There was persistence of short onset heteronymous excitatory responses in triceps brachii, while a normal pattern of restriction of heteronymous responses to pectoralis major and deltoid occurred. The same pattern was observed in older subject groups with spasticity of perinatal origin. In adults with hemiplegia following stroke the threshold of the homonymous phasic stretch reflex was low, but it had a normal onset latency. There was no evidence of abnormal heteronymous excitatory responses. In conclusion, exaggerated excitatory responses to primary muscle afferent input were observed in the homonymous (biceps brachii) and antagonist (triceps brachii) motor neurons in subjects with spasticity arising from perinatal brain damage. They are likely to play an important role in the predominant co-contraction of agonist/antagonist muscles during voluntary movement observed in subjects with spastic cerebral palsy. PMID- 9874489 TI - Dendritic cells enhance the differentiation of naive B cells into plasma cells in vitro. AB - We have shown previously that in vitro-generated human dendritic cells have an effect on the response of B cells at various stages of their differentiation. In a culture system described for the in vitro induction of plasma-cell differentiation, it was reported that naive B cells have a poor propensity to differentiate into plasma cells. In such a culture system, 12% of naive B cells differentiated into plasma cells in the presence of IL-2 and IL-10, despite the interruption of CD40 signalling which is necessary for plasma-cell differentiation. However, as reported herein, naive B cells differentiated fully into plasma cells in response to dendritic cells. Addition of dendritic cells enhanced this differentiation strikingly by recruiting 57% of B cells as plasma cells producing IgM, but also IgG and IgA. In this model, dendritic cells act in synergy with IL-2 at an early stage of CD40-dependent B-cell differentiation, while IL-2 and IL-10 act together, at a later stage, in the generation of plasma cells in a CD40-independent manner. Thus, in addition to the key role played by dendritic cells in the initiation of T-cell responses, our results suggest that dendritic cells regulate humoral responses. PMID- 9874490 TI - Diabetes induction in C57BL/6 mice reconstituted with lymphocytes of nonobese diabetic <--> C57BL/6 mouse embryo aggregation chimeras. AB - To determine whether the genetic background of the insulin-producing beta cells of the pancreas contributes to autoimmune diabetes susceptibility, we have used a model of the disease based on transferring spleen cells from nonobese diabetic (NOD) <--> C57BL/6 (B6) embryo aggregation (EA) chimeras into B6 and NOD irradiated mice. Insulitis and diabetes could be induced into both B6 and NOD hosts, albeit with low incidence. Cyclophosphamide (CY) treatment, known to accelerate diabetes in prediabetic NOD mice, was found to increase diabetes incidence up to 50-60% in both B6 and NOD mice reconstituted with chimeric splenocytes, while diabetes did not occur in CY-treated B6 mice reconstituted with B6 splenocytes. We conclude that the genetic make-up of the target organ does not affect the final stage of the pathogenesis of insulin-dependent diabetes mellitus. PMID- 9874491 TI - Secreted proteins of Mycobacterium leprae. AB - In mycobacteria, secreted proteins represent a distinct group, probably of particular importance for development of immune responses following infection. Quantification of individual proteins in culture fluid and corresponding disrupted bacilli permits determination of a localization index for identification of secreted proteins. This procedure cannot be applied to Mycobacterium leprae because secreted proteins are lost during isolation of bacilli from tissues. The DNA sequences of secreted proteins of Mycobacterium tuberculosis were compared with sequences of M. leprae. Genes for homologues of the 85a, 85b, 85c, mpt32 (apa), mpt51, erp, mtc28, Rv2376c, Rv3354 and Rv0526 genes were identified. All of these contain signal sequences typical for secretion in M. leprae. In several instances the local distance between marker genes and occurrence on the same or the complementary DNA strand was similar in these two species. The genomic organisation of genes for secreted proteins is thus very similar in M. leprae and M. tuberculosis, the homology being higher for the mature polypeptide chains than for the corresponding signal peptides. PMID- 9874492 TI - Influence of intestinal microflora on murine bone marrow and spleen macrophage precursors. AB - To investigate the adjuvant effect of intestinal flora on macrophage-colony stimulating factor-responsive macrophage progenitors from spleen and bone marrow, we compared progenitor numbers and phenotypic characteristics of in vitro matured macrophages in germ-free and flora-associated mice (conventional, Escherichia coli-monoassociated and conventionalized mice). The data obtained show that the flora affected differentially bone marrow and spleen progenitors. It increased the numbers of progenitors in the spleen but not in the bone marrow. It did not modify the expression of F4/80, Mac-1 and major histocompatibility complex (MHC) class II on bone-marrow-derived macrophages (BMDM), while it clearly up-regulated MHC class II expression on spleen-derived macrophages (SDM). This effect was more pronounced in flora-associated ex germ-free mice than in conventional mice and it was greatly enhanced in the absence of M-CSF. In vitro stimulation by lipopolysaccharide had no effect on marker expression of BMDM, while it decreased F4/80 and enhanced MHC class II molecules on SDM from germ-free and flora associated mice. However, the expression of MHC class II remained lower in germ free mice. Enhancement of MHC class II molecule expression on SDM may contribute to the protective role of flora, because successful immune responses are dependent on the expression of these molecules. PMID- 9874493 TI - Oligoclonality of TCRint cells with a low diversity of TCR complementarity determining region 3 in mice with graft-versus-host disease. AB - Conventional T cells (i.e. TCRhigh) are generated by the main stream of T-cell differentiation in the thymus. However, primordial T cells (i.e. TCRint) are generated by extrathymic pathways and an alternative intrathymic pathway. Since TCRint cells contain self-reactive clones, the diversity of the T-cell antigen receptor (TCR) complementarity-determining region (CDR) 3 was examined. The predominant Vbeta8.2+ clones among TCRint cells were selected for DNA sequencing. Thymectomized, irradiated mice subjected to bone-marrow transplantation (BMT) were used; graft-versus-host disease (GVHD), B6-->(B6xC3H/He)F1 and syngeneic BMT, B6-->B6. In these combinations, only TCRint cells were generated. Vbeta8.2+ cells with a low diversity of CDR3 of V-gene expanded in GVHD mice. Vbeta8.2+ cells of TCRint and TCRhigh cells in normal mice were polyclonal, showing that the former has a lower diversity of CDR3 than the latter. The clonality of activated TCRhigh cells was examined, in which CD3high cells (bml2 mice) were injected into 1 Gy-irradiated B6 nude mice. Some Vbeta8.2+ clones among TCRhigh cells were expanding but the diversity of CDR3 was greater than that of CD3int cells, despite the fact that the recognition site of the H-2 difference was smaller. Taken together with invariant usage of V alpha14, these results suggest that TCRint cells have a low diversity of CDR3 of Vbeta genes. PMID- 9874494 TI - Protection against concanavalin A-induced murine liver injury by the organic germanium compound, propagermanium. AB - Propagermanium (3-oxygermylpropionic acid polymer) is an organic germanium compound that activates the immune system. In this study, we investigated the action of propagermanium on T-cell-mediated murine hepatic injury induced by concanavalin A (Con A). Oral administration of propagermanium inhibited the development of liver injury about 10 h after ConA injection. Histological analysis demonstrated that propagermanium attenuated the extent of liver damage compared with controls, reducing infiltration by leucocytes, especially CD11b positive cells. Infiltration by CD4-positive cells was not affected. Tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma are crucial for the development of hepatitis in this model. Propagermanium treatment induced significant inhibition of subsequent TNF-alpha production about 10 h after Con A injection, without affecting IFN-gamma, interleukin (IL)-10, IL-4 and IL-12 production. This effect on TNF-production coincided with the inhibition of aminotransferase activity late in the progression of Con A-induced liver injury. These facts suggest that this compound affects the macrophages (Mphi) function in the liver sinusoid. Therefore, Mphi were cultured with liver sinusoidal endothelial cells (SEC) and the effect of propagermanium on TNF-alpha production in the presence of IFN-gamma was determined. TNF-alpha production was reduced significantly in the coculture of Mphi and SEC when Mphi was treated with propagermanium. These results might explain the mechanisms by which propagermanium inhibits Con-A-induced liver injury. That is, propagermanium improves hepatitis through mechanisms including the reduced production of TNF alpha, without modification of Th1- and Th2-cell function. PMID- 9874495 TI - Tyrosine phosphorylation of p72syk induced by anti-9-O-acetyl GD3 antibodies in human peripheral blood mononuclear cells. AB - We have previously shown that a subpopulation of peripheral blood CD4+ T-cells are strongly positive for the ganglioside 9-O-acetyl GD3. Treatment of peripheral blood mononuclear cells (PBMC) with monoclonal anti-9-O-acetyl GD3 antibodies (MoAb 27A) induced a dose-dependent proliferative response in the cells. Here we show that binding of 27A IgG to PBMC induces rapid tyrosine phosphorylation of p72syk and mobilization of phosphoinositides. Equal phosphorylation was not seen after stimulation of PBMC with anti-GD3 antibodies (MoAb R24) despite the apparent similar cell surface distribution of the two gangliosides. This suggests that the O-acetyl modification enhances the signal transduction mediated by GD3 ganglioside. PMID- 9874496 TI - Coordinate up-regulation of the beta-chemokine subfamily in autoimmune sialoadenitis of MRL/lpr mice. AB - Mononuclear cell (MNC) infiltration of the salivary and lacrimal glands is a major feature in Sjogren's syndrome (SS) and its animal model, murine autoimmune sialoadenitis (MAS). To investigate factors that influence selective infiltration by MNC of submandibular glands in young and adult MRL/lpr mice with MAS, expression of mRNA encoding the beta-chemokines monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta and regulated upon activation, normal T-cell expressed and secreted (RANTES) was investigated by in situ hybridization. MCP-1 protein production was also evaluated by immunohistochemistry. Young mice with MAS showed an early up-regulation of mRNA expression for MCP-1, MIP-1beta and RANTES, while MIP-1alpha mRNA expression was not affected. Adult mice with MAS showed a further up-regulation of mRNA expression for MCP-1, MIP-1beta and RANTES, and a remarkably strong up-regulation for MIP-1alpha. Immunohistochemistry revealed that MCP-1 protein production paralleled MCP-1 mRNA expression in both young and adult mice. These observations implicate MCP-1, MIP-1beta and RANTES as potential chemokines in induction of MAS, and MCP-1, MIP-1beta, RANTES and prominently MIP-1alpha in progression and perturbation of MAS. PMID- 9874498 TI - Costimulatory function of CD28 in avian gammadelta T cells is evolutionarily conserved. AB - CD28 costimulatory signals are required for T-cell proliferation and lymphokine production. In this work, the functional conservation of CD28 was studied in avian gammadelta T cells. The avian CD28 molecule is expressed on all alphabeta T cells and is capable of giving a costimulatory signal. Most peripheral gammadelta T cells are CD28 negative; however, we identified a CD28-positive gammadelta T cell subset from peripheral blood comprising about 12% of gammadelta T cells. The peripheral CD28+ gammadelta T-cell subset included all CD8+ gammadelta T cells known to be a responding subset during activation. After polyclonal activation, the frequency of CD28+ gammadelta T cells was increased and the activation also up-regulated CD5, CD25 and major histocompatibility complex (MHC) class II molecules. These changes were detected after both polyclonal and antigen-specific T-cell activation. In addition, we also showed that CD28 can give a costimulatory signal to gammadelta T cells and that this signal leads to up-regulation of IL-2 and bcl-x transcripts. These results indicate that the function of CD28 is evolutionarily conserved and can already be detected in avian gammadelta T cells. PMID- 9874497 TI - Effect of allergen-specific immunotherapy on interleukin-4, interleukin-5 and interferon-gamma mRNA expression in the nasal mucosa of rats with allergic rhinitis. AB - To elucidate the mechanism of immunotherapy, we tested the effect of ovalbumin and ovalbumin-pullulan conjugate immunotherapy on the expression of interleukin (IL)-4, IL-5 and interferon-gamma (IFN-gamma) mRNA in the nasal mucosa of sensitized rats. Forty-five rats were injected with ovalbumin intraperitoneally on three consecutive days and later were exposed to ovalbumin aerosol. The rats were injected intradermally, on six consecutive days, with saline, ovalbumin or ovalbumin-pullulan conjugate. Later, nasal mucosa was obtained and reverse transcription-polymerase chain reaction (RT-PCR) was performed. Nasal responses and specific immunoglobulin E (IgE) were measured. Although the immunotherapy significantly decreased nasal airway resistance, dye leakage and histamine content in nasal irrigation after allergen challenge, no significant difference was found in IL-4 and IL-5 mRNA expression or in specific IgE level among the three groups. We conclude that in this allergic model, the improvement of nasal responses after immunotherapy was the result of a mechanism other than decrease of T-helper 2 (Th2) cytokines. PMID- 9874499 TI - Assessment of the role of leucocyte function-associated antigen-1 in homotypic adhesion of activated B lymphocytes. AB - In this study we investigated how T-cell-dependent stimuli, via interleukin-4 (IL 4) or CD40 ligation, influence homotypic B-cell adhesion when compared with induction by the T-cell-independent stimulus lipopolysaccharide (LPS). Using primary murine B cells, we found that T-cell-dependent stimulation led to increased aggregation as compared to that induced by LPS. The adhesion was to a large extent dependent on the adhesion molecule, lymphocyte function-associated antigen-1 (LFA-1). We found that activation of B cells with the mitogenic stimuli induced an increased avidity of LFA-1 for its ligand, intercellular adhesion molecule-1 (ICAM-1). The increase was stable and different from that induced by phorbol esters. Although adhesion was reduced using B cells from LFA-1(-/-) mice, aggregation occurred in response to T-cell-dependent stimuli. Our data suggest that adhesion of B lymphocytes is regulated in different modes. One is induced by antigen and leads to a transient conformational change of the LFA-1 molecule. Another is induced by mitogenic stimuli and leads to stable avidity increase of LFA-1, possibly via activation of cytoskeletal anchorage. A third is LFA-1 independent, of low avidity and is induced by T-cell-dependent stimuli. PMID- 9874500 TI - Independent duplications of Bf and C3 complement genes in the zebrafish. AB - As part of an ongoing project aimed at the characterization of the MHC system in bony fishes, we have attempted to identify the class III region in the zebrafish (a teleost), a region that in higher vertebrates contains genes coding for complement proteins C2, Bf and C4. We obtained several genomic PAC clones by hybridization with a zebrafish Bf probe, previously identified in our laboratory, and searched these for the presence of other class III genes. We were able to obtain a second Bf-like gene, however, we were unable to detect any C2- or C4 like genes. By using highly degenerated primers, we extended our search to a hepatopancreas cDNA library and amplified from it clones corresponding to three different C3-like genes, and also the Bf genes, but not any C2- or C4-like genes. The zebrafish therefore contains two Bf and three C3 loci but apparently no C2 and C4 loci. Independent duplications of the Bf and C3 genes in bony fishes suggest that complement plays a prominent part in the immune response of this class of vertebrates. PMID- 9874502 TI - Inhibition of HLA-DR expression on activated human blood eosinophils by transforming growth factor-beta1. AB - Peripheral blood, bronchoalveolar lavage and sputum eosinophils of patients with asthma but not peripheral blood eosinophils from normal controls have been shown to express human leucocyte antigen (HLA)-DR on their cell surface. Cytokines implicated in the activation of eosinophils, such as interleukin (IL)-3 and granulocyte-macrophage colony-stimulating factor (GM-CSF), can up-regulate HLA-DR expression. However, little is known about antagonistic factors that might down regulate HLA-DR expression on eosinophils. In this study we investigated whether transforming growth factor-beta (TGF-beta), which has been shown to reduce survival of activated eosinophils, can also modulate HLA-DR expression on eosinophils. For this purpose, isolated peripheral blood eosinophils were stimulated with IL-3 and GM-CSF for 24 h and HLA-DR expression was measured by flow cytometry. We found that while isolated eosinophils expressed low levels of surface HLA-DR, incubation with GM-CSF and IL-3 increased HLA-DR expression on eosinophils. TGF-beta alone did not change HLA-DR expression on isolated eosinophils. However, co-incubation of eosinophils with TGF-beta and either GM CSF or IL-3 significantly decreased HLA-DR expression compared to eosinophils incubated with either GM-CSF or IL-3 alone and this was not reversed by addition of IL-5. This effect of TGF-beta on IL-3-induced HLA-DR expression was attenuated dose-dependently in the presence of monoclonal anti-TGF-beta antibodies. Our results suggest that TGF-beta can reduce cytokine-induced HLA-DR expression on eosinophils and could thus influence eosinophil activation. PMID- 9874501 TI - Establishment and characterization of a human T-lymphocyte cell line immortalized by SV40 and with abnormal expression of TCR/CD3. AB - Human lymphocytes derived from the peripheral blood of a healthy woman were transfected with a plasmid carrying the simian virus 40 (SV40) large T antigen. The successfully transformed cells contained SV40 large T DNA and were negative for Epstein-Barr virus (EBV) and human T-cell leukaemia virus (HTLV)-1 genomes. The immortalized cell line was assigned to the T-lymphocyte lineage on the basis of morphological, immunological and cytochemical criteria. While the cells expressed CD1a and CD4 at the cell surface, the CD3 complex was solely intracytoplasmic. Immunoprecipitation studies indicated that these cells lacked T cell receptor (TCR) alpha-chains but not beta-chains. They were negative for activation markers such as CD25, CD69 and major histocompatibility (MHC) class II molecules. In addition, the transformed cells exhibited a complete growth independency towards interleukin-2 (IL-2). However, after phorbol ester stimulation, CD25 and CD69 markers were expressed and IL-2 was secreted. This new human immortalized T-lymphocytic cell line, which is cell-surface TCR/CD3 negative, may be useful as an in vitro model for studying TCR/CD3 assembly, expression and signal transduction. PMID- 9874503 TI - Small cytoplasmic antigens from Pseudomonas aeruginosa stimulate gammadelta T lymphocytes. AB - Gammadelta T lymphocytes respond to different bacterial antigens and transformed cells. The antigenic molecules responsible for this activity have been studied extensively in antigenic preparations from Mycobacterium. We describe here the in vitro effect of Pseudomonas aeruginosa on gammadelta T lymphocytes and the properties of the implicated compounds. We found a preferential gammadelta T-cell expansion when we used heat-treated P. aeruginosa preparations and a dose dependent inhibition of proliferation of peripheral blood mononuclear cells (PBMC) when non-heat-treated antigens were studied. This expansion corresponded to a Vgamma9-positive subpopulation. In contrast to alphabeta T lymphocytes, the highest stimulatory activity was restricted to very small cytosolic compounds. This activity was protease resistant and phosphatase sensitive and always dependent on interleukin (IL)-2 or alphabeta T-cell activation. We concluded that the antigenic molecules from P. aeruginosa that activated gammadelta T lymphocytes were small, non-peptidic, phosphorylated compounds, similar to those previously described from Mycobacterium. PMID- 9874504 TI - Distribution of alloreactivity amongst the CD45 isoforms of circulating CD4 and CD8 T lymphocytes. AB - The expression of different isoforms of the CD45 surface molecule allows lymphocytes to be divided into two nonoverlapping categories, CD45RA and CD45RO. Previous studies of CD4 T cells have shown that responses to soluble antigens are present predominantly in the RO subset and to mitogens in the RA, alloreactivity being present in both subsets. Responses of CD8 T cells have not been investigated in such detail, nor have responses been compared to CD4 cells. Here we report the alloreactive responses of both CD45RA and RO subsets amongst both CD4 and CD8 T lymphocytes. Following isolation of CD4 and CD8 cells with immunomagnetic beads, CD45 subsets were separated by negative depletion using specific monoclonal antibodies. CD45RA populations were greater than 97% pure and CD45RO cells greater than 91%. One-way primary mixed lymphocyte reactions were established using the purified responder cells with irradiated allogeneic peripheral blood mononuclear cells as stimulators; experiments were all repeated at least three times. In assays of CD4+ RA and RO subsets, reactivity was present in both isoforms, being consistently, but not significantly, greater amongst the RO subset. With CD8+ T cells, reactivity was also present in both isoforms, but was significantly greater in the CD45RA subset, with mean proliferation 2.5-3 fold that of the CD45RO cells (P < 0.05). PMID- 9874508 TI - Mechanism involved in gallium-67 (Ga-67) uptake by human lymphoid cell lines. AB - The gallium-67 (Ga-67) is a radionuclide used for diagnostic imaging. It is known for its ability to accumulate in inflammatory lesions and tumors, especially in lymphomas. The intracellular distribution and the uptake mechanism of Ga-67 remains a subject of discussion. In this work, we studied the mechanism of Ga-67 uptake in 4 human lymphoid cell lines: 38658, Jurkat, DG75 and U715. Our results have pointed out the heterogeneity of Ga-67 uptake among these cell lines. Using flow cytometry analysis, we reported the expression of transferrin-receptor (TfR) in each lymphoid cell line, and found a positive correlation between TfR density and the Ga-67 uptake (r = 0.99) in lymphoid cells. The anti-TfR monoclonal antibody (MoAb) blocked significantly the Ga-67 uptake in all lymphoid cell lines indicating the Ga-67 uptake as a TfR-dependent mechanism. This mechanism has been saturated with different cold gallium concentrations. A total inhibition of Ga-67 uptake was obtained at a 10(-4) M cold gallium concentration. To conclude, we have shown that the Ga-67 uptake is TfR-dependent in lymphoid cells. Thus, the gallium penetrates in lymphoid cells by an active mechanism which can be saturated. PMID- 9874507 TI - Dysregulation of trace element composition in ovariectomized cynomolgus monkey bones. AB - One of the challenging issues in modern biomedical science is the increasing number of osteoporosis patients due to the expansion of elderly populations. Among aging-related pathogenic changes, alterations in bone function and skeletal pathogenesis is a particularly important issue of concern. Osteoporosis is one of the most serious bone-related pathogenic states, as it causes serious loss of quality of life. Alterations in estrogen levels in accordance with aging are one of the key risk factors for osteoporosis. Complexed estrogen actions on bones can be traced by analyzing bone mineral components, as those elements accumulate as mineral complexes, reflecting the context of multiple cellular reactions such as bone resorption/osteogenesis. We have analyzed bone trace element composition in ovariectomized (OVX-treated) Cynomolgus monkey models in this study. In order to gain insights into the effects of such defects on bone trace element composition, inductively coupled plasma atomic emissions spectrometry (ICP-AES) analysis was performed. Marked changes in bone trace element levels were found in vertebral bones of OVX-treated Cynomolgus monkeys. An assessment of these trace element spectra in OVX model animals is discussed. These results could provide useful markers for understanding the physiological states of bones in postmenopausal women. PMID- 9874509 TI - Modification of Ca2+, Mg2+-ATPase and F-actin distribution in hepatocytes of cyclosporine A treated rats. Effect of soyabean lecithin and triacylglycerol. AB - We studied the effect of cyclosporine A on hepatic Ca2+, Mg2+-ATPase and F-actin on bile canalicular and basolateral membranes in rats fed either soyabean lecithin, or triacylglycerol enriched diet, or low fat diet. Ca2+, Mg2+-ATPase histochemical activity was not modified in lecithin-cyclosporine A group, whereas the activity was decreased in the other groups. The triacylglycerol-cyclosporine A group had the lower activity. The histochemical staining of F-actin was quite normal in lecithin-cyclosporine group but decreased in the other cyclosporine A treated groups. The lower staining was observed in the triacylglycerol cyclosporine group. The alteration of Ca2+, Mg2+-ATPase and F-actin by cyclosporine A, related to cholestasis evidenced by a decrease in bile salt secretion, were prevented by dietary soyabean lecithin and amplified by dietary soyabean triacylglycerol. PMID- 9874510 TI - Increased cyclin E level in retinoblastoma cells during programmed cell death. AB - Camptothecin (an inhibitor of topoisomerase I) and etoposide and amsacrine (inhibitors of topoisomerase II) both capable of triggering programmed cell death in Y79 cells, induced a remarkable dose-dependent increase in the level of cyclin E in these cells. Camptothecin was found to be the most effective compound. The effect was not observed when the cells were treated with other inducers of programmed cell death (C2-ceramide, sodium butyrate, interleukin-1beta and tumor necrosis factor), all of which do not damage DNA. The effect, which was completely prevented by inhibitors of macromolecular synthesis, occurred after a lag phase (12 hrs.) and increased concurrently with the rise in programmed cell death (PCD), reaching a maximum after 36 hrs. of incubation, when a large percentage of cells (95%) showed clear PCD signals. We suggest that cyclin E takes part in the final stage of programmed cell death which is induced by topoisomerase inhibitors in Y79 cells. PMID- 9874511 TI - Modulation of bone morphogenetic protein-2 and bone morphogenetic protein-4 gene expression in osteoblastic cell lines. AB - Bone morphogenetic proteins (BMPs) belong to the transforming growth factor-beta (TGF-beta) superfamily and are crucial factors in the process of bone formation. Despite knowledge on their wide distribution and expression, however, there is very little information on the biological factors that affect gene transcription of these osteoinductive agents. To investigate this aspect of BMP gene regulation we have studied the effect of a number of factors known to affect osteogenic cells. Northern analysis showed modulation of the expression of BMP-2 and BMP-4 mRNAs in two human osteosarcoma cell lines, MG63 and Saos-2, by prostaglandin E2 (PGE2), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interferon-alpha (IFN alpha), retinoic acid and 1,25(OH)2 vitamin D3. mRNA expressions of the normally used "housekeeping genes", glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin, were found to be susceptible to influence by some of the factors used. Hence, an oligo(dT)15-18 probe was used to reliably estimate the relative quantities of mRNA present for normalization of data. In general, all factors down-regulated mRNA expressions of BMP-2 and BMP-4 in MG63 cells. IL-6 completely abolished detectable expression of BMP-2 mRNA, which was also greatly reduced by IL-1beta, retinoic acid and 1,25(OH)2 vitamin D3. PGE2 had similar influences on BMP-2 and BMP-4 expressions, showing reductions to approximately 60% of normal. In Saos-2 cells only 1,25(OH)2 vitamin D3 had any great effect on BMP-2 expression, which was down-regulated to approximately 60% of control values. BMP 4 was down-regulated by IFN-alpha (approximately 60%) and IL-1beta (approximately 20%). We conclude that BMPs are subject to regulation by a variety of factors and that this is dependent on the stage of the cell in the osteogenic lineage. Furthermore, the use of GAPDH and beta-actin genes as "housekeeping genes" in expression-modulation studies must be treated with care. PMID- 9874512 TI - Optimization of the tyrosinase mRNA probe to detect circulating melanocytes with reverse transcription and polymerase chain reaction. AB - It was recently suggested that reverse transcription polymerase chain reaction (RT-PCR)-based detection of tyrosinase messenger RNA (mRNA) in peripheral blood is useful in the early detection of circulating tumor cells, since tyrosinase is thought to be a melanocyte-specific marker. However, the sensitivity of detection of these cells in circulation is controversial, some authors reporting 0% effectiveness, others obtaining 100% efficacy. We developed a modification of a technique to process blood samples to detect tyrosinase mRNA, and tested the method with 50 samples from as many patients with histologically confirmed malignant melanoma in different stages. Whole blood was processed by discarding the plasma and extracting RNA from density gradient-isolated peripheral blood lymphocytes. The RNA samples were tested with a sensitive nested primer RT-PCR assay. Sensitivity was tested using RNA extracted from SK-mel-1 human melanoma cells diluted serially with peripheral blood obtained from healthy control subjects. A lymph node from a patient with confirmed disseminated melanoma served as the positive control. Our technique was able to detect tyrosinase mRNA in samples from the 37 patients with progressive metastatic melanoma. The test detected tyrosinase mRNA from both the melanoma cell line and the positive lymph node. Our method to extract RNA from whole blood improves the specificity and sensitivity of tyrosinase mRNA detection by RT-PCR. The test should be of use in determining the prognosis of patients with melanoma, and in deciding when to initiate early treatment in patients with malignant melanoma. PMID- 9874513 TI - Age-related differences in calcium accumulation in human arteries. AB - To elucidate the accumulation of calcium in the human arteries, the calcium contents of the thoracic aorta, coronary, common carotid, basilar, internal thoracic, axillary, radial, femoral, popliteal, and dorsalis pedis arteries, were analyzed by inductively coupled plasma atomic emission spectrometry (ICP-AES). The calcium content began to increase in both the thoracic aorta and femoral artery around the age of 50 years (yrs), in the popliteal artery at the age of 60 yrs, in the coronary, basilar and dorsalis pedis arteries at the age of 70 yrs, and in the common carotid artery at the age of 80 yrs. In the same time, the calcium content did not increase significantly in the internal thoracic and radial arteries. Accumulation of calcium in human arteries was classified into two groups: The first is an age-related increase of calcium content in the arteries like the thoracic aorta, coronary, common carotid, basilar, axillary, femoral, popliteal and dorsalis pedis arteries. The second is non-age-related, such as the internal thoracic and radial arteries. To examine the localization of this calcium accumulation, the thoracic-aortic and femoral-arterial walls were separated into the three tunicae, intima, media and adventitia. In the case of the thoracic aorta, the accumulation of calcium and phosphorus occurred primarily in the tunica media of aorta, secondarily in the tunica intima. With regard to the femoral artery, the accumulation of calcium and phosphorus occurred only in the tunica media, only in the tunica intima, or in both the tunicae media and intima. Therefore, the manner of accumulation of calcium and phosphorus in the femoral-arterial wall was different from that in the aortic wall. Comparing the upper and lower limb arteries, the calcium content was found to be higher in the femoral, popliteal, and dorsalis pedis arteries of the lower limb than that of the axillary and radial arteries of the upper limb. PMID- 9874514 TI - 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulates the adipose conversion of piglet preadipocytes in primary culture. AB - The effects of 12-O-tetradecanoylphorbol-13-acetate (TPA) were analyzed on the proliferation and differentiation of cultured porcine preadipocytes. In both chemically-defined and serum-containing media, short term treatment (2 days), either during the growth phase or from confluence, significantly enhanced cell proliferation, as assessed by tritiated thymidine incorporation and protein content assay. Addition of TPA during the growth phase in serum-free medium had no effect on lipoprotein lipase (LPL), glycerol 3-phosphate dehydrogenase (GPDH) and malic enzyme (ME) activities after 14 days of culture. By contrast, similar treatments in serum-containing medium significantly increased all these enzyme activities. Addition of TPA from confluence in both media stimulated LPL activity on day 14 as well as ME activity on day 17. These results were confirmed by morphologic studies. In conclusion, this study demonstrates that a short stimulation of the protein kinase C pathway can enhance the differentiation of porcine preadipocytes. Such a positive effect is however dependent on the time of TPA addition as well as the culture conditions, indicating a complex regulation of the protein kinase C pathway in this cell type. PMID- 9874516 TI - Laser-induced autofluorescence spectroscopy of dental caries. AB - We studied spectral autofluorescence characteristics of dental caries. A wide range of carious lesions revealed characteristic emission of endogenous fluorophores with strong fluorescence bands in the red spectral region when excited with 407 nm line radiation of a krypton ion laser. Healthy hard dental tissue exhibited no emission bands in the red. The fluorescence spectra, fluorescence excitation spectra as well as the reflectance spectra of carious lesions were found to be typical for fluorescent porphyrins, mainly protoporphyrin IX. A possible source of these porphyrins within carious tissues is bacterial biosynthesis. Non-invasive sensitive in vivo caries detection by means of appropriate excitation sources and porphyrin fluorescence detectors should be possible. PMID- 9874515 TI - X-rays-induced secretion of cellular factor(s) that enhance(s) HIV-1 promoter transcription in various non-irradiated transfected cell lines. AB - Various cellular stress agents like ionizing radiation exposure could activate human immunodeficiency virus type 1 (HIV- 1) replication or reporter gene expression. In addition, extracellular factor(s) released by X-ray-treated human colonic carcinoma cell line (HT29) might activate the long terminal repeat (LTR) of HIV-1 in non-irradiated HT29 cells. In the present report we show that in various transiently or stably transfected cell lines, X-ray irradiation up regulates HIV-1 LTR transcription through the kappaB regulatory elements. A factor(s), which is processed by and acts upon a variety of cell types, was detected by addition to non-irradiated cells of either X-ray-treated cells or a conditioned medium taken from irradiated cultures. The magnitude of responsiveness is cell type dependent. In addition, X-ray activation of HIV-1 LTR in transiently or stably transfected cell lines is inhibited by a potent antioxidant drug, pyrrolidine dithiocarbamate and by another drug, known for its role in the trapping of growth factors, suramin. The importance of these observations in the pathophysiology of patients with AIDS-related cancers treated by radiotherapy remains to be established. PMID- 9874517 TI - Recovery of motor function after focal cortical injury in primates: compensatory movement patterns used during rehabilitative training. AB - The recovery of skilled hand use after cortical injury was assessed in adult squirrel monkeys. Specific movement patterns used to perform a motor task requiring fine manual skill were analyzed before and after a small ischemic infarct (2.6-3.8 mm2) to the electrophysiologically identified hand area of the primary motor cortex (M1). After 1-3 weeks of pre-infarct training, each monkey stereotypically used one specific movement pattern to retrieve food pellets. After injury to the hand area of M1, the monkeys were required to retrieve the pellets using their impaired forelimb. Immediately after the injury, the number of finger flexions used by the monkeys to retrieve the pellets increased, indicating a deficit in skilled finger use. After approximately 1 month of rehabilitative training, skilled use of the fingers appeared to recover, indicated by a reduction in the number of finger flexions per retrieval. The monkeys again retrieved the pellets using one specific movement pattern in most trials. Despite the apparent recovery of skilled finger use after rehabilitative training, three of five monkeys retrieved the pellets using stereotypic movement patterns different from those used before the injury. Thus, this study provides evidence that compensatory movement patterns are used in the recovery of motor function following cortical injury, even after relatively small lesions that produce mild, transient deficits in motor performance. Examination of electrophysiological maps of evoked movements suggests that the mode of recovery (re-acquisition of pre-infarct movement strategies vs development of compensatory movement strategies) may be related to the relative size of the lesion and its specific location within the M1 hand representation. PMID- 9874518 TI - Neural gain changes subserving perceptual acuity. AB - When small and large objects of equal weight are lifted, the small object feels heavier than the large one (the size-weight illusion) and requires more effort to lift (the size-effort illusion). It has been suggested that these illusions result from neural gain changes designed to maintain acuity under different working conditions. If this suggestion is correct, a given mass should produce a larger increase in perceived weight or effort when added to the small object. This was found to be the case. PMID- 9874519 TI - Examination of the relationships between jaw opener and closer rhythmical muscle activity in an in vitro brainstem jaw-attached preparation. AB - An in vitro jaw-attached brainstem preparation was developed to investigate the relationship between jaw opener and closer muscle activity during chemically induced rhythmical jaw movements in neonatal rats. In the majority of preparations examined, where a defined region of brainstem was isolated and the neuronal innervation of the jaw opener and closer muscles was left intact, bath application of the excitatory amino acid agonist N-methyl-D,L-aspartate (NMA, 20 40 microM) in combination with bicuculline (BIC 10 microM), a GABA(A) antagonist, produced rhythmical electromyogram (EMG) activity in jaw opener and closer muscles, bilaterally, in conjunction with rhythmical jaw movements. Low concentrations of NMA (20 microM) in combination with BIC produced temporally coordinated activity between the jaw opener and closer muscles, ipsilaterally. With higher doses of NMA (40 microM), each muscle group exhibited bursting, but temporal coordination between them was difficult to establish. Similarly, NMA application in combination with the glycine antagonist strychnine (STR, 10 microM), also produced rhythmical EMG activity from both opener and closer muscles, ipsilaterally, but showed no temporal coordination between the antagonist muscle pair. However, coordination of opener and closer muscle discharge could be restored by the addition of BIC to the bath. We suggest that there exist separate, but coordinated, rhythm generator circuits for opener and closer motoneuronal discharge located in close proximity to the trigeminal motor nucleus and under GABAergic control for production of temporal coordination between rhythmogenic circuits. PMID- 9874520 TI - Subdivisions of the motor and somatosensory thalamus of primates revealed with Wisteria floribunda agglutinin histochemistry. AB - We obtained well-differentiated staining of thalamic subdivisions in rhesus macaques and squirrel monkeys using a lectin, Wisteria floribunda agglutinin (WFA), that labels extracellular matrix proteoglycans. Regional variations in staining were observed within the motor and somatosensory thalamic regions that bear on current interpretations of the organization of these regions. The pattern of WFA staining was generally similar to that obtained with Cat-301 antibody, which also stains proteoglycans. However, WFA reliably produced selective staining in both squirrel monkeys and macaques, whereas Cat-301 stained macaques more consistently than squirrel monkeys. PMID- 9874521 TI - The influence of rate of temperature change and peak stimulus duration on pain intensity and quality. AB - An important aspect of experimental pain research is that the assessment methods can investigate the different aspects of pain perception. The aim of the present study was to investigate the influence of rate of temperature change and peak stimulus duration on heat evoked pain intensity and quality. All stimuli were applied within the medial aspect of the anterior forearm. The rate of temperature change was varied from 1 to 16 degrees C/s without any effect on the pain threshold. The pain threshold decreased with an increasing peak stimulus duration from 0.1 to 2 s, but not from 2 to 3 s. The pain intensity for suprathreshold stimuli (46 degrees C, 48 degrees C, 50 degrees C) increased for decreasing rates and increasing duration. The pain intensity was highly correlated with the energy of the stimulus. When the rates of temperature change (1-16 degrees C/s) are varied, no differences between pricking and burning pain were present at either low stimulus intensity (46 degrees C) or high stimulus intensity (50 degrees C). At low stimulus intensity (46 degrees C), the pricking pain was not influenced by the duration (0.1-3 s), but the burning pain was intensified when the duration was increased from 1.5 to 3 s. At high intensity stimuli (50 degrees C), the pricking pain intensified with an increased duration, whereas burning pain did not. The heat pain threshold is influenced by the peak stimulus duration, and not by the rate of temperature change. If suprathreshold stimuli are used, both the rate of temperature change and the peak stimulus duration can strongly affect the pain intensity and the pain quality. Therefore, the same stimulus modality can be used to assess the modulation of different pain intensities and of the pricking and burning pain qualities simply by varying the stimulus configuration. PMID- 9874522 TI - Neurotransmitter system markers in adult barrel field. PMID- 9874523 TI - Inner ear imaging: more than "rule out acoustic". PMID- 9874524 TI - Lymphoma: master of chicanery. PMID- 9874525 TI - Neonatal MR imaging: achieving our own expectations. PMID- 9874526 TI - Subarachnoid hemorrhage due to late recurrence of a previously unruptured aneurysm after complete endovascular occlusion. PMID- 9874527 TI - Radiology managing radiology. PMID- 9874528 TI - FDA medical device regulation and informed consent. PMID- 9874529 TI - Inner ear volumetric measurements using high-resolution 3D T2-weighted fast spin echo MR imaging: initial experience in healthy subjects. AB - BACKGROUND AND PURPOSE: Adult size is achieved in the inner ear labyrinth by approximately 25 weeks' gestation, and minimal variability in age, sex, side, and race is found after birth. In this study, we opted to determine the reproducibility of inner ear volumetric measurements generated from high resolution heavily T2-weighted 3D fast spin-echo MR images. METHODS: The temporal bones of 23 volunteers were imaged using a heavily T2-weighted 3D fast spin-echo MR imaging technique. The images were assessed by a neuroradiologist for the presence of inner ear configurational anomalies and, most important, for complete coverage of the inner ear labyrinth. Subsequently, the volume of the fluid in the inner ear was determined by two observers using a semiautomated segmentation algorithm. The mean, SD, range, and coefficient of variation of fluid volume in the inner ear were calculated. Age-, sex-, and side-related differences in the inner ear volumetric measurements were evaluated using analysis of variance. Interrater consistency in the inner ear volumetric measurements was evaluated by comparing the calculated coefficients of reliability. RESULTS: Volumetric measurements were available from 46 inner ears in 23 volunteers. The mean volume was 227.8 mm3 (SD, 24.4 mm3), and the coefficient of variation was 10.7%. No age , sex-, or side-related differences in the inner ear volumetric measurements were found (F ratios were 4.33, 5.04, and 0.26, respectively). Interrater consistency, as assessed by the coefficient of reliability, was 5.3%. CONCLUSION: Reproducible volumetric measurements of the inner ear labyrinth can be obtained by applying a semiautomated segmentation algorithm to a heavily T2-weighted 3D fast spin-echo MR imaging data set. These volumetric measurements may help identify patients with congenital sensorineural hearing loss and normal inner ear configuration. PMID- 9874530 TI - Delineation of small nerves and blood vessels with three-dimensional fast spin echo MR imaging: comparison of presurgical and surgical findings in patients with hemifacial spasm. AB - BACKGROUND AND PURPOSE: We applied a 3D fast spin-echo (3D-FSE) MR imaging technique to the preoperative and postoperative evaluation of patients with hemifacial spasm. METHODS: The study group comprised 20 patients. All images were acquired on a 1.5-T MR system with a 3D-FSE sequence. RESULTS: In all 20 patients, the courses of the seventh and eighth cranial nerves were depicted separately, and the arteries presumed to be responsible for the hemifacial spasm were seen to be in contact with the facial nerves at the root exit zone (REZ). Eight patients underwent neurovascular decompression. In all patients, the presumed responsible blood vessels depicted by 3D-FSE MR imaging corresponded to intraoperative findings. In addition, postoperative 3D-FSE images confirmed the separation of the facial nerve from a contiguous vessel at the REZ. DISCUSSION: The 3D-FSE technique makes it possible to obtain extremely high-quality images of microstructures in the cerebellopontine cistern, and it has several advantages over conventional angiography: it is noninvasive and able to depict the cranial nerves and surrounding vessels in the same image without contrast material, and it may be useful for postoperative evaluation of the decompression procedure. This imaging technique is expected to prove useful for the clinical evaluation of hemifacial spasm. PMID- 9874531 TI - Entirely intracanalicular meningioma: contrast-enhanced MR findings in a rare entity. AB - We report an unusual case of an entirely intracanalicular meningioma in a 51-year old woman. Contrast-enhanced MR images showed an enhancing lesion filling the lateral portion of the left internal auditory canal. Our findings suggest that meningiomas should be considered in the differential diagnosis of entirely intracanalicular masses. PMID- 9874532 TI - Parotid pleomorphic adenomas: delayed CT enhancement. AB - BACKGROUND AND PURPOSE: Weak or absent CT enhancement in pleomorphic adenomas of the parotid gland has been observed immediately after i.v. contrast administration. This feature can result in poor lesion conspicuity relative to both normal parotid tissue and other parotid abnormalities. The purpose of this study was to document the delayed CT enhancement characteristics of parotid pleomorphic adenomas and to preliminarily compare these results with the enhancement characteristics of other parotid gland tumors. METHODS: Preoperative CT scans from 18 pathologically proved parotid gland neoplasms were reviewed retrospectively. Lesions included eight pleomorphic adenomas, four Warthin's tumors, two squamous cell carcinomas, two mucoepidermoid cancers, one acinic cell carcinoma, and one melanoma metastasis. In all cases, axial CT was performed after the administration of 100 to 150 mL of i.v. contrast material, followed by delayed (average, 24 minutes; range, 13 to 34 minutes) coronal CT scanning. The mean normalized Hounsfield unit (HU) attenuation of each lesion was computed by drawing a region of interest around the entire mass and dividing the resulting HU value by that of the contralateral uninvolved parotid gland. RESULTS: For all eight pleomorphic adenomas, the degree of contrast enhancement increased and became progressively more uniform with time. Mean normalized axial lesion enhancement averaged 1.20 +/- 0.35 at 8 minutes, compared with 2.30 +/- 0.66 on the coronal scans at 24 minutes. For the 10 nonpleomorphic adenomas, no significant change was found in either the degree or pattern of contrast enhancement between the immediate and delayed CT scans. In these tumors, peak enhancement was reached early, during axial scanning. CONCLUSION: Delayed CT contrast enhancement is observed in parotid pleomorphic adenomas, increasing in both degree and homogeneity with time. This feature may be useful in selecting an appropriate contrast delay when scanning possible pleomorphic adenomas to improve lesion conspicuity and, potentially, to better distinguish these tumors from other parotid abnormalities. PMID- 9874533 TI - Internal carotid artery narrowing in children with retropharyngeal lymphadenitis and abscess. AB - BACKGROUND AND PURPOSE: Our purpose was to describe the association between narrowing of the internal carotid artery (ICA) and retropharyngeal abscess in children. METHODS: Neck CT scans from 13 consecutive children with suppurative retropharyngeal lymphadenitis and abscess were evaluated retrospectively for asymmetric ICA diameters at the level of the abscess. Clinical status at the time of illness was established via a chart review. Twenty control CT scans obtained from pediatric patients with normal imaging findings were evaluated prospectively to determine symmetry and size of the ICA. RESULTS: Mean diameter of the normal ICA, contralateral to the retropharyngeal abscess, was 5 mm (range, 3-8 mm), while mean diameter ipsilateral to the abscess was 3 mm (range, 1-5 mm). The diameters of the normal and abnormal ICAs were statistically significantly different. All children were neurologically normal. The right and left ICAs in children with normal CT findings in the neck were symmetrical in diameter. CONCLUSION: Despite dramatic narrowing of the ICA ipsilateral to retropharyngeal lymphadenitis and abscess, no children in this series had neurologic deficits, suggesting that such narrowing is a common, benign, and, most likely, incidental imaging finding. PMID- 9874534 TI - Isolated vagal nerve palsy associated with a dissection of the extracranial internal carotid artery. AB - A 40-year-old man had paralysis of the right vocal cord. Imaging showed a dissection of the extracranial internal carotid artery, and physical examination disclosed paresis of the right side of the soft palate. To our knowledge, this is only the second report of carotid dissection presenting as an isolated vagal neuropathy. Most often, multiple lower cranial nerves are involved. The CT, MR imaging, and MR angiographic findings are presented and the topic is reviewed. PMID- 9874535 TI - Extranodal non-Hodgkin lymphoma involving the muscles of mastication. AB - Involvement of the skeletal muscle by non-Hodgkin lymphoma (NHL) is unusual. The most commonly affected muscles are those of the extremities, pelvis, and gluteal regions; rarely are the muscles in the head and neck involved. We report the clinical, CT, and MR imaging findings in two patients with extranodal NHL in the head and neck region involving the muscles of mastication. One patient was immunocompetent and had lymphoma that arose within the muscles; the other was a patient with AIDS who had disseminated disease at diagnosis. In both patients, the involved muscles were isodense with normal muscles on CT scans. On MR images, the infiltrated muscles were isointense with normal muscles on the T1-weighted sequence and hyperintense on the fast spin-echo T2-weighted sequence, with variable enhancement after administration of intravenous contrast material. PMID- 9874536 TI - Amyloidoma of the gasserian ganglion. AB - We report an unusual case of an amyloidoma of the gasserian ganglion associated with trigeminal neuralgia. MR imaging showed a mass in Meckel's cave, which was isointense with surrounding tissue on T1-weighted images and hypointense on T2 weighted images. After contrast administration, the mass enhanced homogeneously, and thin cuts through the region showed involvement in the expected location of the gasserian ganglion and, more laterally and inferiorly, in the proximal part of V1. A review of the literature revealed that only one similar case has been reported since the advent of modern neuroimaging. Amyloidoma, although rare, may be considered as a rare differential diagnosis of a mass in this region. PMID- 9874537 TI - MR outcome parameters in multiple sclerosis: comparison of surface-based thresholding segmentation and magnetization transfer ratio histographic analysis in relation to disability (a preliminary note). AB - BACKGROUND AND PURPOSES: MR imaging is now widely used to monitor disease progression in patients with multiple sclerosis (MS). The purpose of this study was to explore the relationship between disability status and existing and new MR parameters in MS patients. METHODS: Forty-one patients with clinically definitive MS were studied. MR imaging included T2- and T1-weighted imaging as well as gradient-echo imaging with and without magnetization transfer contrast. We used surface-based thresholding segmentation techniques to obtain T2 and T1 lesion load, T1/T2 ratio, and several magnetization transfer ratio (MTR) lesion load parameters. MTR histographic analysis included measurements of absolute peak height (aHp), relative peak height (rHp), MTR of the peak (MTRp), mean MTR (MTRm), and MTR25, MTR50, and MTR75, relating to the integrals of the histogram at 25%, 50%, and 75%, respectively, of the total area under the curve. All MR parameters were correlated with Expanded Disability Status Scale (EDSS) score, disease duration, and patient's age. RESULTS: Using surface-based thresholding segmentation techniques, we found relatively low correlations with EDSS. T1 lesion load and T1/T2 ratios correlated most strongly. Regarding MTR histographic parameters, EDSS correlated best with rHp but only weakly with others. Similar correlations were found with disease duration, but not with age. CONCLUSION: The best MR correlations with disability were several MTR histographic parameters. Our findings may favor the use of these MR parameters over T2 lesion load to monitor disease progression in patients with MS, findings that should be explored further in longitudinal studies. PMID- 9874538 TI - Sensitivity of enhanced MR in multiple sclerosis: effects of contrast dose and magnetization transfer contrast. AB - BACKGROUND AND PURPOSES: New strategies have been developed to improve the sensitivity of contrast-enhanced MR imaging in quantifying disease activity in patients with multiple sclerosis (MS). The goal of the present study was to evaluate the sensitivity of T1-weighted images after injection of a triple dose of contrast material and application of a magnetization transfer (MT) pulse in the detection of enhancing lesions as compared with the conventional approach. METHODS: Monthly MR images were obtained in 13 patients with relapsing-remitting MS for a period of 3 months. The MR studies were performed on two separate occasions with single- and triple-dose contrast material. In each session, T1- and T2-weighted spin-echo images with and without the MT pulse were obtained before and after contrast administration. All images were evaluated in a blinded fashion and scored in random order and consensually by two readers. The number of total and new enhancing lesions and active images was counted. RESULTS: Eighty six percent more enhancing lesions and 54% more new enhancing lesions were detected with triple-dose as compared with single-dose non-MT sequences, whereas single-dose MT images depicted 33% more enhancing lesions and 18% more new enhancing lesions than the single-dose non-MT images. Twenty-nine percent more lesions were detected on triple-dose non-MT images than on single-dose MT images. The combination of a triple dose of contrast material and MT did not produce any significant change in detection of enhancing lesions as compared with a triple dose of contrast without MT. CONCLUSION: The use of a triple dose of contrast material is the best approach to maximize the sensitivity of enhanced MR imaging. PMID- 9874539 TI - A brain image database for structure/function analysis. AB - BACKGROUND AND PURPOSE: Lesion-deficit-based structure-function analysis has traditionally been empirical and nonquantitative. Our purpose was to establish a new brain image database (BRAID) that allows the statistical correlation of brain functional measures with anatomic lesions revealed by clinical brain images. METHODS: Data on 303 participants in the MR Feasibility Study of the Cardiovascular Health Study were tested for lesion/deficit correlations. Functional data were derived from a limited neurologic examination performed at the time of the MR examination. Image data included 3D lesion descriptions derived from the MR examinations by hand segmentation. MR images were normalized in-plane using local, linear Talairach normalization. A database was implemented to support spatial data structures and associated geometric and statistical operations. The database stored the segmented lesions, patient functional scores, and several anatomic atlases. Lesion-deficit association was sought by contingency testing (chi2-test) for every possible combination of each neurologic variable and each labeled atlas structure. Significant associations that confirmed accepted lesion-deficit relationships were sought. RESULTS: Two-hundred thirty-five infarctlike lesions in 117 subjects were viewed collectively after mapping into Talairach cartesian coordinates. Anatomic structures most strongly correlated with neurologic deficits tended to be situated in anatomically appropriate areas. For example, infarctlike lesions associated with visual field defects were correlated with structures in contralateral occipital structures, including the optic radiations and occipital gyri. CONCLUSION: Known lesion deficit correlations can be established by a database using a standard coordinate system for representing spatial data and incorporating functional and structural data together with appropriate query mechanisms. Improvements and further applications of this methodology may provide a powerful technique for uncovering new structure-function relationships. PMID- 9874540 TI - Proton MR spectroscopic findings correspond to neuropsychological function in traumatic brain injury. AB - BACKGROUND AND PURPOSE: Traumatic brain injury (TBI) causes substantial irreversible damage to neurons. Our aim was to investigate whether proton MR spectroscopic measures of diffuse cellular integrity were related to neuropsychological dysfunction after TBI. METHODS: Twelve patients with TBI (mean, 53 +/- 23 days postinjury) and 14 control subjects were included in the study using paired MR spectroscopy and neuropsychological assessment. N acetylaspartate (NAA), creatine, and choline were measured in normal-appearing occipitoparietal white and occipital gray matter using short-echo quantitative spectroscopy. A composite measure of neuropsychological function was calculated from z-scored individual tests probing the major functional domains commonly impaired after head trauma. RESULTS: Patients with TBI displayed reduced NAA in white matter and elevated choline in gray matter, suggestive of neuronal injury and inflammation, respectively. NAA and creatine in white and gray matter were significantly associated with composite neuropsychological function and many individual neuropsychological tests. Gray matter choline, although abnormal, was not related to neuropsychological function. CONCLUSION: The concordance between neurometabolic levels and behavioral function supports the hypothesis that diffuse axonal injury is an important contributor to brain dysfunction after TBI. PMID- 9874541 TI - Ophthalmoplegic migraine: reversible enhancement and thickening of the cisternal segment of the oculomotor nerve on contrast-enhanced MR images. AB - BACKGROUND AND PURPOSE: Ophthalmoplegic migraine is a rare condition characterized by the association of headaches and an oculomotor nerve palsy. We report six patients with typical clinical features of this disorder in whom enhancement of the cisternal segment of the oculomotor nerve developed during the acute phase, followed by resolution of enhancement over several weeks as the symptoms resolved. METHODS: Six patients, ages 3 to 27 years, underwent MR imaging during the acute phase of ophthalmoplegic migraine and at the time of recovery several weeks later. The MR studies, performed on a 1.5-T MR unit, included noncontrast and contrast-enhanced axial and coronal T1-weighted sequences. Sagittal images were obtained in two patients, with and without contrast enhancement. RESULTS: Enhancement of the cisternal segment of the oculomotor nerve was seen in all patients at initial presentation. Contrast enhanced studies also showed focal thickening at the exit of the nerve in the interpeduncular cistern in five of six patients. No patient had enhancement of the cavernous sinus or adjacent dura. Enhancement was almost completely resolved on follow-up studies 7 to 9 weeks later. CONCLUSION: Our findings confirm an intrinsic transient abnormality in the cisternal segment of the third nerve in patients with a typical clinical presentation of ophthalmoplegic migraine. PMID- 9874542 TI - Diagnosis of middle cerebral artery stenosis by transcranial color-coded real time sonography. AB - BACKGROUND & PURPOSE: This study was performed to determine the usefulness of transcranial color-coded real-time sonography (TCCS) in detecting stenosis in the horizontal portion of the middle cerebral artery (MCA). METHODS: Using TCCS and the incident angle correction technique, we measured the peak-systolic flow velocity in bilateral MCAs in 45 consecutive patients in whom cerebral angiography was carried out within 1 week before or after TCCS. Three patients had a stenosis of 75% or greater and four had a unilateral occlusion of the extracranial internal carotid artery (ICA) (the ICS and ICO groups, respectively). Eight patients had a stenosis of 50% or greater (one bilateral and seven unilateral) (the M1S group). Four patients had unilateral distal occlusion of the horizontal portion of the MCA (the M1O group). Twenty-six patients had no significant extra- or intracranial stenosis on the ipsilateral or contralateral side (the control group). RESULTS: Mean peak-systolic flow velocity on the affected side was 83.0 +/- 20.8 cm/s in the ICS group, 59.8 +/- 23.2 cm/s in the ICO group, and 62.3 +/- 33.7 cm/s in the M1O group. In the control group, the mean peak-systolic flow velocity was 116.0 +/- 31.5 cm/s. In the M1S group, however, the mean peak-systolic flow velocity (334.2 +/- 35.7 cm/s) on the affected side always exceeded 180 cm/s (mean value +/- 2 SD in the control group), and was significantly higher than that in the other groups. The mean peak systolic flow velocity in the M1S group increased with the grade of stenosis. CONCLUSION: The M1S group members could easily be distinguished from the other group members by their peak-systolic flow velocity in excess of 180 cm/s. Measurement of the peak-systolic flow velocity of the MCA by TCCS may help to identify a significant stenosis in the horizontal portion of the MCA. PMID- 9874543 TI - Multifocal varicella-zoster virus leukoencephalitis in a patient with AIDS: MR findings. AB - We describe a patient with AIDS who presented with an acute encephalitis caused by infection with varicella-zoster virus. The hemorrhagic, necrotizing encephalitis had an unusual MR appearance, with innumerable discrete, small, targetlike lesions in the right cerebral hemisphere, which were coalescent in the posterior temporal, parietal, and occipital regions. Of the several known disease patterns of varicella-zoster viral infection in the CNS, this histopathologic pattern of multifocal leukoencephalitis is rare. It is important to recognize, as effective antiviral drug treatments are available. PMID- 9874544 TI - Rhombencephalitis caused by adenovirus: MR imaging appearance. AB - Encephalitis is a rare manifestation of adenovirus infection. We report the MR imaging findings of a patient with rhombencephalitis caused by adenovirus. Imaging findings included T2 signal abnormalities in the brain stem and cerebellum with mild patchy enhancement and mass effect. PMID- 9874545 TI - Dentato-rubral tract involvement in adult-onset adrenoleukodystrophy. PMID- 9874546 TI - MR imaging technique for the diagnosis of pituitary iron overload in patients with transfusion-dependent beta-thalassemia major. AB - To identify the optimal MR imaging technique for diagnosing pituitary iron overload, we compared spin-echo and gradient-echo MR imaging with measurements of pituitary T2 relaxation times in 30 patients with secondary hemochromatosis due to transfusion-dependent beta-thalassemia major and in 10 healthy volunteers. We found that the optimal MR imaging technique to evaluate pituitary iron overload is the gradient-echo T2*-weighted technique, as it best demonstrated signal reduction in the anterior lobe of the pituitary gland. PMID- 9874547 TI - MR imaging of perinatal brain damage: comparison of clinical outcome with initial and follow-up MR findings. AB - BACKGROUND AND PURPOSE: The purpose of our study was to determine whether MR studies in the neonatal period are predictive of the neuroradiologic sequelae and clinical outcome in premature and term infants with perinatal brain injury. METHODS: Thirty subjects (15 premature and 15 term infants) with abnormalities revealed by initial MR studies were reexamined approximately 1 year after birth with both MR imaging and a neurologic assessment. All initial MR studies were performed between 35 and 45 weeks corrected age in premature infants and within 28 days of life in term infants. The initial MR studies were evaluated for deep gray matter involvement, hemispheric parenchymal change, intracranial hemorrhage, and periventricular signal and/or morphologic changes. These MR findings were compared with the follow-up MR findings and with the neurologic outcome. RESULTS: The development of cerebral palsy in premature infants was related to the following initial MR findings: subependymal hemorrhage associated with parenchymal destruction, periventricular signal alteration with irregularity of the ventricular wall, and widespread cerebral infarction. These MR findings were predictive of the subtypes of cerebral palsy. In term asphyxiated infants, T2 signal alterations of the deep gray matter rather than T1 shortening and diffuse involvement of the hemispheres were predictive of an unfavorable outcome. Both in term and premature infants, focal hemispheric parenchymal lesions alone (including infarction and intracerebral, subdural, intraventricular, and subarachnoid hemorrhage) did not produce poor outcomes. CONCLUSION: MR studies performed at or near term in either premature or term infants with perinatal brain damage are effective in predicting both late neuroradiologic and clinical outcome. PMID- 9874548 TI - Age-dependent changes in magnetization transfer contrast of white matter in the pediatric brain. AB - BACKGROUND AND PURPOSE: It is unknown to what extent magnetization transfer contrast (MTC) in white matter of the brain changes during myelination. The goal of this study was to measure the age-dependent changes of MTC in different regions of the pediatric brain and to evaluate their relation to T2 relaxation times. METHODS: Seventy children aged 1 week to 80 months without evidence of organic brain disease underwent MR imaging of the brain. A double-echo spin-echo (SE) sequence and an SE sequence with and without an off-resonance pulse were performed in the axial orientation. Using paired images, we calculated MTC ratios in 13 predefined regions of the brain and compared them with the T2 relaxation times measured in the same areas. Regression analysis was performed for both parameters to evaluate age dependency. RESULTS: MTC in white matter increased during myelination from a range of 13% to 19% to a range of 34% to 37%. At the same time, T2 relaxation times decreased from a range of 115 to 160 milliseconds to a range of 60 to 70 milliseconds after myelination. For both MTC and T2 relaxation times, age dependency could be expressed by a monoexponential function. CONCLUSION: A strong positive correlation exists between MTC ratios and the degree of myelination in the pediatric brain, and an inverse correlation exists between MTC and T2 relaxation times. Fast proton relaxation within macromolecules in the myelinated white matter and subsequent MT may be the most important reason for the decreasing T2 relaxation time of white matter during brain myelination. PMID- 9874549 TI - Sonographic ventriculography: a new potential use for sonographic contrast agents in neonatal hydrocephalus. AB - BACKGROUND AND PURPOSE: Sonographic identification of communicating and noncommunicating forms of posthemorrhagic hydrocephalus can be difficult. This study evaluates the feasibility of using sonographic contrast agents to determine ventricular patency in a newborn animal model. METHODS: A craniotomy was performed and a catheter was placed under sonographic guidance into the lateral ventricle in five anesthetized newborn piglets. Sonograms were obtained before and after intraventricular injection of 0.01 mL of a sonographic contrast agent (Imagent, formulation AF0150; Alliance Pharmaceutical Corp, San Diego, CA) diluted in 1 mL normal saline, and images were assessed for presence and location of echogenic contrast material. RESULTS: Flow of contrast material was identifiable from the ipsilateral lateral ventricle into the contralateral lateral ventricle, and through the third and fourth ventricles into the basal cisterns during real-time sonography in every animal. Ventricular and cistern echogenicity remained increased for approximately 4 minutes after injection. CONCLUSION: Contrast-enhanced sonographic ventriculography has the potential to identify patency of CSF pathways in newborns with hydrocephalus and an indwelling ventricular catheter. PMID- 9874550 TI - Hemimegalencephaly and focal megalencephaly in tuberous sclerosis complex. AB - We describe two children with complex cortical malformations as well as the typical intracranial manifestations of tuberous sclerosis complex. One child had hemimegalencephaly and the other had extensive focal megalencephaly. These cases are discussed in terms of the current concepts of cortical malformations. PMID- 9874551 TI - Subarachnoid hemorrhage due to late recurrence of a previously unruptured aneurysm after complete endovascular occlusion. AB - We present a case of subarachnoid hemorrhage attributed to rupture of an aneurysm 18 months after endovascular occlusion. The aneurysm was diagnosed after the patient had a seizure; however, there was no evidence of subarachnoid hemorrhage. Angiography at 6 months revealed a totally occluded aneurysm. This case illustrates that the long-term results of endovascular occlusion remain uncertain. PMID- 9874552 TI - Reversible ischemia determined by xenon-enhanced CT after 90 minutes of complete basilar artery occlusion. AB - Intraarterial thrombolytic therapy decreases mortality in the treatment of acute basilar artery occlusion. An acute decrease in cerebral blood flow (CBF) (<12 mL/100 g per minute) has been reported to invariably result in infarction. We report a case of acute basilar artery occlusion, recanalized within 90 minutes, with reversal of CBF of less than 6 mL/100 g per minute. After reperfusion, areas with persistent CBF of 6 mL/100 g per minute resulted in infarctions on subsequent CT studies. Parenchymal viability is possible after 90 minutes of posterior CBF of 6 mL/100 g per minute. PMID- 9874553 TI - Imaging the pathophysiology of infarction in the clinical setting. PMID- 9874554 TI - Stenting of symptomatic stenosis of the intracranial internal carotid artery. AB - Percutaneous balloon angioplasty has been reported to be useful in the treatment of intracranial atherosclerotic arterial stenoses; however, arterial dissection with increased risk of stroke has limited its widespread implementation. Although vascular stenting has improved the outcome of similar procedures in the extracranial vasculature, its intracranial application has been limited by the lack of suitable stents. We report a case of successful percutaneous stenting of a symptomatic intracranial carotid artery using a second-generation, flexible, coronary stent. PMID- 9874555 TI - Coil embolization of a trigeminal-cavernous fistula. AB - A 53-year-old woman spontaneously incurred a right trigeminal artery-cavernous sinus fistula, manifested by an intracranial bruit and right sixth nerve palsy. This lesion was successfully managed by coil embolization via the transvenous and transarterial routes. PMID- 9874556 TI - Percutaneous laser photocoagulation of spinal osteoid osteomas under CT guidance. AB - BACKGROUND AND PURPOSE: Spinal osteoid osteomas are rare; when they occur, they are usually treated by surgical or percutaneous excision. The aim of percutaneous interstitial laser photocoagulation (ILP) of osteoid osteomas under CT guidance is thermal destruction of the nidus using low-power laser energy, thus precluding bone resection and open surgery. METHODS: Three cases of spinal osteoid osteomas were treated with percutaneous ILP of the nidus. Under CT guidance, the needle was positioned in the center of the nidus, at least 8 mm from neurologic structures. Using a high-power semiconductor diode laser (805 nm) with a 400 microm optical fiber, we delivered 600 to 800 joules to the nidus, depending on its size. The procedure was performed with the patient under neuroleptanalgesia and required overnight hospitalization. RESULTS: Complete pain relief was obtained in all three patients within 24 hours of the procedure, and no major complications were incurred. Follow-up ranged from 20 months to 60 months. CONCLUSION: Percutaneous ILP of spinal osteoid osteoma is a promising, simple, precise, and minimally invasive technique and may be an alternative to traditional surgical and percutaneous ablations. PMID- 9874557 TI - Postradiation colospinal fistula: a rare cause of spinal epidural abscess. AB - We describe a rare case of spinal epidural abscess, which resulted from a colospinal fistula that developed as a complication of previous surgical resection and radiotherapy for rectal carcinoma. The patient's clinical course and the imaging features of colospinal fistula are described, along with a brief review of the literature. PMID- 9874558 TI - Direct emergence of the dorsospinal artery from the aorta supplying the anterior spinal artery: report of two cases. AB - We report two cases of an isolated dorsospinal artery that emerged directly from the aorta to supply the anterior spinal artery, which in one case had another blood supply on the opposite side, in a lower position. It is important to identify this anatomic variant, otherwise spinal angiography might be incomplete, especially when the isolated dorsospinal artery supplies the anterior spinal artery. PMID- 9874559 TI - Human non-germinal center B cell interleukin (IL)-12 production is primarily regulated by T cell signals CD40 ligand, interferon gamma, and IL-10: role of B cells in the maintenance of T cell responses. AB - Interleukin (IL)-12 is expressed mainly in antigen-presenting cells after challenge with microbial material or after CD40 activation. Although IL-12 was cloned from human Epstein-Barr virus (EBV)-transformed B cell lines, surprisingly, CD40 ligation on murine B cells did not lead to IL-12 production, suggesting that murine B cells do not produce IL-12. Here we demonstrate that a subset of human tonsillar B cells can be induced to express and secrete bioactive IL-12. The major stimulus to produce IL-12 in human B cells was CD40 ligation. In contrast, B cell receptor cross-linking did not induce IL-12. Expression of IL-12 after CD40 activation was restricted to CD38(-)IgD+/- non-germinal center (non GC) B cells. CD40 ligation and interferon (IFN)-gamma exhibited synergistic effects on IL-12 production, whereas IL-10 abrogated and IL-4 significantly inhibited IL-12 production by these B cells. In contrast to IL-12, production of IL-6 is conversely regulated, leading to significant increase after CD40 ligation in the presence of the T helper type 2 (Th2) cytokine IL-4. Cord blood T cells skewed towards either a Th1 or a Th2 phenotype maintained their cytokine expression pattern when restimulated with allogeneic resting B cells. Blockade of CD40 and/or IL-12 during T-B interaction significantly reduced IFN-gamma production by the T cells. This suggests a model whereby B cells produce either IL-12 or IL-6 after contact with T cells previously differentiated towards Th1 or Th2. Furthermore, IL-12 and IL-6 might provide a positive feedback during cognate T-B interactions, thereby maintaining T cells' differentiation pattern during amplification of the immune response. PMID- 9874560 TI - A T cell receptor-specific blockade of positive selection. AB - To investigate the influence of endogenous peptides on the developmental processes that occur during thymocyte selection, we have used monoclonal antibodies that preferentially recognize the major histocompatibility complex (MHC) molecule I-Ek when it is bound to the moth cytochrome c peptide (88-103). One of these antibodies (G35) specifically blocks the positive selection of transgenic thymocytes expressing a T cell receptor that is reactive to this peptide- MHC complex. Furthermore, G35 does not block the differentiation of transgenic T cells bearing receptors for a different I-Ek-peptide complex. This antibody recognizes a subset of endogenous I-Ek-peptide complexes found on a significant fraction of thymic antigen-presenting cells, including cortical and medullary epithelial cells. The sensitivity of G35 to minor alterations in peptide sequence suggests that the thymic peptide-MHC complexes that mediate the positive selection of a particular class II MHC-restricted thymocyte are structurally related to the complexes that can activate it in the periphery. PMID- 9874561 TI - Immune adherence-mediated opsonophagocytosis: the mechanism of Leishmania infection. AB - To mimic the sandfly pool feeding process and characterize the cellular and biochemical events that occur during the early stages of promastigote-host interaction, we developed an ex vivo model of human blood infection with Leishmania promastigotes. Within 30 s of blood contact, Leishmania promastigotes bind natural anti-Leishmania antibodies, which then activate the classical complement pathway and opsonization by the third component of complement. The opsonized promastigotes undergo an immune adherence reaction and bind quantitatively to erythrocyte CR1 receptors; opsonized Leishmania amastigotes also bind to erythrocytes. Progression of infection implies promastigote transfer from erythrocytes to acceptor blood leukocytes. After 10 min of ex vivo infection, 25% of all leukocytes contain intracellular parasites, indicating that blood cells are the early targets for the invading promastigotes. We propose that adaptation to the immune adherence mechanism aids Leishmania survival, promoting rapid promastigote phagocytosis by leukocytes. This facilitates host colonization and may represent the parasite's earliest survival strategy. In light of this mechanism, it is unlikely that infection-blocking vaccines can be developed. PMID- 9874562 TI - L-selectin-mediated leukocyte adhesion in vivo: microvillous distribution determines tethering efficiency, but not rolling velocity. AB - Adhesion receptors that are known to initiate contact (tethering) between blood borne leukocytes and their endothelial counterreceptors are frequently concentrated on the microvilli of leukocytes. Other adhesion molecules are displayed either randomly or preferentially on the planar cell body. To determine whether ultrastructural distribution plays a role during tethering in vivo, we used pre-B cell transfectants expressing L- or E-selectin ectodomains linked to transmembrane/intracellular domains that mediated different surface distribution patterns. We analyzed the frequency and velocity of transfectant rolling in high endothelial venules of peripheral lymph nodes using an intravital microscopy model. Ectodomains on microvilli conferred a higher efficiency at initiating rolling than random distribution which, in turn, was more efficient than preferential expression on the cell body. The role of microvillous presentation was less accentuated in venules below 20 micrometers in diameter than in larger venules. In the narrow venules, tethering of cells with cell body expression may have been aided by forced margination through collision with erythrocytes. L selectin transfected cells rolled 10-fold faster than E-selectin transfectants. Interestingly, rolling velocity histograms of cell lines expressing equivalent copy numbers of the same ectodomain were always similar, irrespective of the topographic distribution. Our data indicate that the distribution of adhesion receptors has a dramatic impact on contact initiation between leukocytes and endothelial cells, but does not play a role once rolling has been established. PMID- 9874563 TI - The HIV-1 virion-associated protein vpr is a coactivator of the human glucocorticoid receptor. AB - The HIV-1 virion-associated accessory protein Vpr affects both viral replication and cellular transcription, proliferation, and differentiation. We report that Vpr enhances the activity of glucocorticoids in lymphoid and muscle-derived cell lines by interacting directly with the glucocorticoid receptor and general transcription factors, acting as a coactivator. Vpr contains the signature motif LXXLL also present in cellular nuclear receptor coactivators, such as steroid receptor coactivator 1 and p300/CREB-binding protein, which mediates their interaction with the glucocorticoid and other nuclear hormone receptors. A mutant Vpr molecule with disruption of this coactivator signature motif lost its ability to influence transcription of glucocorticoid-responsive genes and became a dominant-negative inhibitor of Vpr, possibly by retaining its general transcription factor-binding activities. The glucocorticoid coactivator activity of Vpr may contribute to increased tissue glucocorticoid sensitivity in the absence of hypercortisolism and to the pathogenesis of AIDS. PMID- 9874564 TI - STAT3 is required for the gp130-mediated full activation of the c-myc gene. AB - The signal transducers and activators of transcription (STAT) family members have been implicated in regulating the growth, differentiation, and death of normal and transformed cells in response to either extracellular stimuli, including cytokines and growth factors, or intracellular tyrosine kinases. c-myc expression is coordinately regulated by multiple signals in these diverse cellular responses. We show that STAT3 mostly mediates the rapid activation of the c-myc gene upon stimulation of the interleukin (IL)-6 receptor or gp130, a signal transducing subunit of the receptor complexes for the IL-6 cytokine family. STAT3 does so most likely by binding to cis-regulatory region(s) of the c-myc gene. We show that STAT3 binds to a region overlapping with the E2F site in the c-myc promoter and this site is critical for the c-myc gene promoter- driven transcriptional activation by IL-6 or gp130 signals. This is the first identification of the linkage between a member of the STAT family and the c-myc gene activation, and also explains how the IL-6 family of cytokines is capable of inducing the expression of the c-myc gene. PMID- 9874565 TI - Rearrangement and expression of immunoglobulin light chain genes can precede heavy chain expression during normal B cell development in mice. AB - In mouse mutants incapable of expressing mu chains, VkappaJkappa joints are detected in the CD43(+) B cell progenitors. In agreement with these earlier results, we show by a molecular single cell analysis that 4-7% of CD43(+) B cell progenitors in wild-type mice rearrange immunoglobulin (Ig)kappa genes before the assembly of a productive VHDHJH joint. Thus, mu chain expression is not a prerequisite to Igkappa light chain gene rearrangements in normal development. Overall, approximately 15% of the total CD43(+) B cell progenitor population carry Igkappa gene rearrangements in wild-type mice. Together with the results obtained in the mouse mutants, these data fit a model in which CD43(+) progenitors rearrange IgH and Igkappa loci independently, with a seven times higher frequency in the former. In addition, we show that in B cell progenitors VkappaJkappa joining rapidly initiates kappa chain expression, irrespective of the presence of a mu chain. PMID- 9874566 TI - Identification and characterization of novel superantigens from Streptococcus pyogenes. AB - Three novel streptococcal superantigen genes (spe-g, spe-h, and spe-j) were identified from the Streptococcus pyogenes M1 genomic database at the University of Oklahoma. A fourth novel gene (smez-2) was isolated from the S. pyogenes strain 2035, based on sequence homology to the streptococcal mitogenic exotoxin z (smez) gene. SMEZ-2, SPE-G, and SPE-J are most closely related to SMEZ and streptococcal pyrogenic exotoxin (SPE)-C, whereas SPE-H is most similar to the staphylococcal toxins than to any other streptococcal toxin. Recombinant (r)SMEZ, rSMEZ-2, rSPE-G, and rSPE-H were mitogenic for human peripheral blood lymphocytes with half-maximal responses between 0.02 and 50 pg/ml (rSMEZ-2 and rSPE-H, respectively). SMEZ-2 is the most potent superantigen (SAg) discovered thus far. All toxins, except rSPE-G, were active on murine T cells, but with reduced potency. Binding to a human B-lymphoblastoid line was shown to be zinc dependent with high binding affinity of 15-65 nM. Evidence from modeled protein structures and competitive binding experiments suggest that high affinity binding of each toxin is to the major histocompatibility complex class II beta chain. Competition for binding between toxins was varied and revealed overlapping but discrete binding to subsets of class II molecules in the hierarchical order (SMEZ, SPE-C) > SMEZ-2 > SPE-H > SPE-G. The most common targets for the novel SAgs were human Vbeta2.1- and Vbeta4-expressing T cells. This might reflect a specific role for this subset of Vbetas in the immune defense of gram-positive bacteria. PMID- 9874567 TI - Distinct subsets of CD1d-restricted T cells recognize self-antigens loaded in different cellular compartments. AB - Although recent studies have indicated that the major histocompatibility complex like, beta2-microglobulin-associated CD1 molecules might function to present a novel chemical class of antigens, lipids and glycolipids, to alpha/beta T cells, little is known about the T cell subsets that interact with CD1. A subset of CD1d autoreactive, natural killer (NK)1.1 receptor-expressing alpha/beta T cells has recently been identified. These cells, which include both CD4(-)CD8(-) and CD4(+) T cells, preferentially use an invariant Valpha14-Jalpha281 T cell receptor (TCR) alpha chain paired with a Vbeta8 TCR beta chain in mice, or the homologous Valpha24-JalphaQ/Vbeta11 in humans. This cell subset can explosively release key cytokines such as interleukin (IL)-4 and interferon (IFN)-gamma upon TCR engagement and may regulate a variety of infectious and autoimmune conditions. Here, we report the existence of a second subset of CD1d-restricted CD4(+) T cells that do not express the NK1.1 receptor or the Valpha14 TCR. Like the Valpha14(+) NK1.1(+) T cells, these T cells exhibit a high frequency of autoreactivity to CD1d, use a restricted albeit distinct set of TCR gene families, and contribute to the early burst of IL-4 and IFN-gamma induced by intravenous injection of anti-CD3. However, the Valpha14(+) NK1.1(+) and Valpha14(-) NK1.1(-) T cells differ markedly in their requirements for self antigen presentation. Antigen presentation to the Valpha14(+) NK1.1(+) cells requires endosomal targeting of CD1d through a tail-encoded tyrosine-based motif, whereas antigen presentation to the Valpha14(-) NK1.1(-) cells does not. These experiments suggest the existence of two phenotypically different subsets of CD1d restricted T cells that survey self-antigens loaded in distinct cellular compartments. PMID- 9874568 TI - Cooperative inhibition of T-cell antigen receptor signaling by a complex between a kinase and a phosphatase. AB - Antigen receptor-triggered T-cell activation is mediated by the sequential action of the Src and Syk/Zap-70 families of protein tyrosine kinases (PTKs). Previously, we reported that another PTK termed p50(csk) was a potent negative regulator of T-cell receptor (TCR) signaling because of its ability to inactivate Src-related kinases. This inhibitory effect required the catalytic activity of Csk, as well as its Src homology (SH)3 and SH2 domains. Subsequent studies uncovered that, via its SH3 domain, p50(csk) was associated with PEP, a proline enriched protein tyrosine phosphatase (PTP) of unknown function expressed in hemopoietic cells. Herein, we have attempted to identify the role of the Csk-PEP complex in T lymphocytes. The results of our experiments showed that, like Csk, PEP was a strong repressor of TCR signaling. This property was dependent on the phosphatase activity of PEP, as well as on the sequence mediating its binding to p50(csk). Through reconstitution experiments in Cos-1 cells, evidence was obtained that Csk and PEP act synergistically to inhibit protein tyrosine phosphorylation by Src-related kinases, and that this effect requires their association. Finally, experiments with a substrate-trapping mutant of PEP suggested that PEP functions by dephosphorylating and inactivating the PTKs responsible for T-cell activation. In addition to giving novel insights into the mechanisms involved in the negative regulation of T-cell activation, these findings indicate that the association of an inhibitory PTK with a PTP constitutes a more efficient means of inhibiting signal transduction by Src family kinases in vivo. PMID- 9874569 TI - Activated Ras signals developmental progression of recombinase-activating gene (RAG)-deficient pro-B lymphocytes. AB - To elucidate the intracellular pathways that mediate early B cell development, we directed expression of activated Ras to the B cell lineage in the context of the recombination-activating gene 1 (RAG1)-deficient background (referred to as Ras RAG). Similar to the effects of an immunoglobulin (Ig) mu heavy chain (HC) transgene, activated Ras caused progression of RAG1-deficient progenitor (pro)-B cells to cells that shared many characteristics with precursor (pre)-B cells, including downregulation of surface CD43 expression plus expression of lambda5, RAG2, and germline kappa locus transcripts. However, these Ras-RAG pre-B cells also upregulated surface markers characteristic of more mature B cell stages and populated peripheral lymphoid tissues, with an overall phenotype reminiscent of B lineage cells generated in a RAG- deficient background as a result of expression of an Ig mu HC together with a Bcl-2 transgene. Taken together, these findings suggest that activated Ras signaling in pro-B cells induces developmental progression by activating both differentiation and survival signals. PMID- 9874570 TI - Mitochondria-dependent and -independent regulation of Granzyme B-induced apoptosis. AB - Granzyme B (GraB) is required for the efficient activation of apoptosis by cytotoxic T lymphocytes and natural killer cells. We find that GraB and perforin induce severe mitochondrial perturbation as evidenced by the release of cytochrome c into the cytosol and suppression of transmembrane potential (Deltapsi). The earliest mitochondrial event was the release of cytochrome c, which occurred at the same time as caspase 3 processing and consistently before the activation of apoptosis. Granzyme K/perforin or perforin treatment, both of which kill target cells efficiently but are poor activators of apoptosis in short term assays, did not induce rapid cytochrome c release. However, they suppressed Deltapsi and increased reactive oxygen species generation, indicating that mitochondrial dysfunction is also associated with this nonapoptotic cell death. Pretreatment with peptide caspase inhibitors zVAD-FMK or YVAD-CHO prevented GraB apoptosis and cytochrome c release, whereas DEVD-CHO blocked apoptosis but did not prevent cytochrome c release, indicating that caspases act both up- and downstream of mitochondria. Of additional interest, Deltapsi suppression mediated by GraK or GraB and perforin was not affected by zVAD-FMK and thus was caspase independent. Overexpression of Bcl-2 and Bcl-XL suppressed caspase activation, mitochondrial cytochrome c release, Deltapsi suppression, and apoptosis and cell death induced by GraB, GraK, or perforin. In an in vitro cell free system, GraB activates nuclear apoptosis in S-100 cytosol at high doses, however the addition of mitochondria amplified GraB activity over 15-fold. GraB- induced caspase 3 processing to p17 in S-100 cytosol was increased only threefold in the presence of mitochondria, suggesting that another caspase(s) participates in the mitochondrial amplification of GraB apoptosis. We conclude that GraB-induced apoptosis is highly amplified by mitochondria in a caspase-dependent manner but that GraB can also initiate caspase 3 processing and apoptosis in the absence of mitochondria. PMID- 9874571 TI - An essential role for nuclear factor kappaB in promoting double positive thymocyte apoptosis. AB - To examine the role of nuclear factor (NF)-kappaB in T cell development and activation in vivo, we produced transgenic mice that express a superinhibitory mutant form of inhibitor kappaB-alpha (IkappaB-alphaA32/36) under the control of the T cell-specific CD2 promoter and enhancer (mutant [m]IkappaB-alpha mice). Thymocyte development proceeded normally in the mIkappaB-alpha mice. However, the numbers of peripheral CD8(+) T cells were significantly reduced in these animals. The mIkappaB-alpha thymocytes displayed a marked proliferative defect and significant reductions in interleukin (IL)-2, IL-3, and granulocyte/macrophage colony-stimulating factor production after cross-linking of the T cell antigen receptor. Perhaps more unexpectedly, double positive (CD4(+)CD8(+); DP) thymocytes from the mIkappaB-alpha mice were resistant to alpha-CD3-mediated apoptosis in vivo. In contrast, they remained sensitive to apoptosis induced by gamma-irradiation. Apoptosis of wild-type DP thymocytes after in vivo administration of alpha-CD3 mAb was preceded by a significant reduction in the level of expression of the antiapoptotic gene, bcl-xL. In contrast, the DP mIkappaB-alpha thymocytes maintained high level expression of bcl-xL after alpha CD3 treatment. Taken together, these results demonstrated important roles for NF kappaB in both inducible cytokine expression and T cell proliferation after TCR engagement. In addition, NF-kappaB is required for the alpha-CD3-mediated apoptosis of DP thymocytes through a pathway that involves the regulation of the antiapoptotic gene, bcl-xL. PMID- 9874572 TI - Mature follicular dendritic cell networks depend on expression of lymphotoxin beta receptor by radioresistant stromal cells and of lymphotoxin beta and tumor necrosis factor by B cells. AB - The formation of germinal centers (GCs) represents a crucial step in the humoral immune response. Recent studies using gene-targeted mice have revealed that the cytokines tumor necrosis factor (TNF), lymphotoxin (LT) alpha, and LTbeta, as well as their receptors TNF receptor p55 (TNFRp55) and LTbetaR play essential roles in the development of GCs. To establish in which cell types expression of LTbetaR, LTbeta, and TNF is required for GC formation, LTbetaR-/-, LTbeta-/-, TNF /-, B cell-deficient (BCR-/-), and wild-type mice were used to generate reciprocal or mixed bone marrow (BM) chimeric mice. GCs, herein defined as peanut agglutinin-binding (PNA+) clusters of centroblasts/centrocytes in association with follicular dendritic cell (FDC) networks, were not detectable in LTbetaR-/- hosts after transfer of wild-type BM. In contrast, the GC reaction was restored in LTbeta-/- hosts reconstituted with either wild-type or LTbetaR-/- BM. In BCR-/ recipients reconstituted with compound LTbeta-/-/BCR-/- or TNF-/-/BCR-/- BM grafts, PNA+ cell clusters formed in splenic follicles, but associated FDC networks were strongly reduced or absent. Thus, development of splenic FDC networks depends on expression of LTbeta and TNF by B lymphocytes and LTbetaR by radioresistant stromal cells. PMID- 9874573 TI - DNA vaccination: transfection and activation of dendritic cells as key events for immunity. AB - The mechanisms underlying initiation and maintenance of CD4 T cell responses after DNA vaccination were studied using a construct coding for nonsecreted fifth component of complement (C5) protein, thus restricting the availability of antigen. The only cell types to express C5 were keratinocytes at the site of DNA application and a small number of dendritic cells present in the draining lymph nodes. Antigen expression persisted for up to 12 wk in keratinocytes, but dendritic cells did not express C5 beyond 2 wk after vaccination. Cross-priming of dendritic cells by C5 expressed in keratinocytes did not occur unless keratinocyte death was induced by irradiation in vitro. CD4 T cells were activated in the draining lymph nodes only and subsequently migrated to the spleen, where memory T cells persisted for longer than 40 wk despite the absence of a source of persistent antigen. While DNA vaccination resulted in transfection of a small proportion of dendritic cells only, it led to general activation of all dendritic cells, thus providing optimal conditions for effective T cell activation and maintenance of memory. PMID- 9874574 TI - Modulation of immune complex-induced inflammation in vivo by the coordinate expression of activation and inhibitory Fc receptors. AB - Autoantibodies and immune complexes are major pathogenic factors in autoimmune injury, responsible for initiation of the inflammatory cascade and its resulting tissue damage. This activation results from the interaction of immunoglobulin (Ig)G Fc receptors containing an activation motif (ITAM) with immune complexes (ICs) and cytotoxic autoantibodies which initiates and propagates an inflammatory response. In vitro, this pathway can be interrupted by coligation to FcgammaRIIB, an IgG Fc receptor containing an inhibitory motif (ITIM). In this report, we describe the in vivo consequences of FcgammaRII deficiency in the inflammatory response using a mouse model of IC alveolitis. At subthreshold concentrations of ICs that fail to elicit inflammatory responses in wild-type mice, FcgammaRII deficient mice developed robust inflammatory responses characterized by increased hemorrhage, edema, and neutrophil infiltration. Bronchoalveolar fluids from FcgammaRII-/- stimulated mice contain higher levels of tumor necrosis factor and chemotactic activity, suggesting that FcgammaRII deficiency lowers the threshold of IC stimulation of resident cells such as the alveolar macrophage. In contrast, complement- and complement receptor-deficient mice develop normal inflammatory responses to suprathreshold levels of ICs, while FcRgamma-/- mice are completely protected from inflammatory injury. An inhibitory role for FcgammaRII on macrophages is demonstrated by analysis of FcgammaRII-/- macrophages which show greater phagocytic and calcium flux responses upon FcgammaRIII engagement. These data reveal contrasting roles for the cellular receptors for IgG on inflammatory cells, providing a regulatory mechanism for setting thresholds for IC sensitivity based on the ratio of ITIM to ITAM FcgammaR expression. Exploiting the FcgammaRII inhibitory pathway could thus provide a new therapeutic approach for modulating antibody-triggered inflammation. PMID- 9874575 TI - Deletion of fcgamma receptor IIB renders H-2(b) mice susceptible to collagen induced arthritis. AB - Autoimmune diseases, like rheumatoid arthritis, result from a dysregulation of the immune response culminating in hyperactivation of effector cells leading to immune-mediated injury. To maintain an appropriate immune response and prevent the emergence of autoimmune disease, activation signals must be regulated by inhibitory pathways. Biochemical and genetic studies indicate that the type IIB low-affinity receptor for immunoglobulin (Ig)G (FcgammaRIIB) inhibits cellular activation triggered through antibody or immune complexes and may be an important component in preventing the emergence of autoimmunity. To investigate the role of FcgammaRIIB in the development of type II collagen (CII)-induced arthritis (CIA), a model for rheumatoid arthritis in humans, we have examined its contribution in determining the susceptibility to CIA in the nonpermissive H-2(b) haplotype. H 2(b) mice immunized with bovine CII do not develop appreciable disease. In contrast, immunization of the FcgammaRIIB-deficient, H-2(b) mice with bovine CII induced CIA at an incidence of 42.2%. The maximal arthritis index of the FcgammaRIIB-deficient mice developing CIA (6.9 +/- 3.6) was comparable to that of DBA/1 mice (8.6 +/- 1.9), an H-2(q) strain susceptible for CIA induction. IgG1, IgG2a, and IgG2b antibody responses against CII were elevated in the FcgammaRIIB deficient animals, especially in those mice showing arthritis, but less pronounced than DBA/1 mice. Histological examinations of the arthritic paws from FcgammaRIIB-deficient mice revealed that cartilage was destroyed and bone was focally eroded in association with marked lymphocyte and monocyte/macrophage infiltration, very similar to the pathologic findings observed in DBA/1 mice. These results indicate that a nonpermissive H-2(b) haplotype can be rendered permissive to CIA induction through deletion of FcgammaRIIB, suggesting that FcgammaRIIB plays a critical role in suppressing the induction of CIA. PMID- 9874576 TI - Molecular recognition of lipid antigens by T cell receptors. AB - The T cell antigen receptor (TCR) mediates recognition of peptide antigens bound in the groove of major histocompatibility complex (MHC) molecules. This dual recognition is mediated by the complementarity-determining residue (CDR) loops of the alpha and beta chains of a single TCR which contact exposed residues of the peptide antigen and amino acids along the MHC alpha helices. The recent description of T cells that recognize hydrophobic microbial lipid antigens has challenged immunologists to explain, in molecular terms, the nature of this interaction. Structural studies on the murine CD1d1 molecule revealed an electrostatically neutral putative antigen-binding groove beneath the CD1 alpha helices. Here, we demonstrate that alpha/beta TCRs, when transferred into TCR deficient recipient cells, confer specificity for both the foreign lipid antigen and CD1 isoform. Sequence analysis of a panel of CD1-restricted, lipid-specific TCRs reveals the incorporation of template-independent N nucleotides that encode diverse sequences and frequent charged basic residues at the V(D)J junctions. These sequences permit a model for recognition in which the TCR CDR3 loops containing charged residues project between the CD1 alpha helices, contacting the lipid antigen hydrophilic head moieties as well as adjacent CD1 residues in a manner that explains antigen specificity and CD1 restriction. PMID- 9874579 TI - VII International Congress of Reproductive Immunology. New Delhi, India, October 27-30, 1998. Abstracts. PMID- 9874580 TI - Analyzing protein families and domains on the Web. PMID- 9874582 TI - Biomedical research in cardiovascular diseases: still searching for the fountain of youth. PMID- 9874578 TI - Leptin is an endogenous protective protein against the toxicity exerted by tumor necrosis factor. AB - Tumor necrosis factor (TNF) is a central mediator of a number of important pathologies such as the systemic inflammatory response syndrome. Administration of high TNF doses induces acute anorexia, metabolic derangement, inflammation, and eventually shock and death. The in vivo effects of TNF are largely mediated by a complex network of TNF-induced cytokines and hormones acting together or antagonistically. Since TNF also induces leptin, a hormone secreted by adipocytes that modulates food intake and metabolism, we questioned the role of leptin in TNF-induced pathology. To address this question, we tested mouse strains that were defective either in leptin gene (ob/ob) or in functional leptin receptor gene (db/db), and made use of a receptor antagonist of leptin. Ob/ob and db/db mice, as well as normal mice treated with antagonist, exhibited increased sensitivity to the lethal effect of TNF. Exogenous leptin afforded protection to TNF in ob/ob mice, but failed to enhance the protective effect of endogenous leptin in normal mice. We conclude that leptin is involved in the protective mechanisms that allow an organism to cope with the potentially autoaggressive effects of its immune system. PMID- 9874581 TI - Cell to cell contact and extracellular matrix. PMID- 9874583 TI - Mechanisms of horizontal gene spreading. Proceedings of a workshop. Oslo, Norway, June 8-9, 1998. PMID- 9874584 TI - New biological markers may accurately predict prognosis in head and neck cancer patients. PMID- 9874585 TI - Novel form of Vitamin K may stop liver cancer cell growth. PMID- 9874586 TI - Workshop on Intratumoral Aromatase. Winterthur, Switzerland, 10-11 July 1997. PMID- 9874587 TI - Non-simultaneous deaths of parallel personhoods crashing through a Denver S & L. PMID- 9874588 TI - Loss, humiliation and entrapment as appraisals of schizophrenic illness: a prospective study of depressed and non-depressed patients. AB - OBJECTIVES: Life-events that precede the onset of unipolar depression usually involve an appraisal of loss; recent research has shown that where these events are also appraised as humiliating or involving entrapment and defeat (the absence of a way forward or failure to reaffirm an identity) they are especially potent in triggering depression. Depression in schizophrenia has not been studied from the cognitive or psychosocial perspectives, probably because of its confused nosological status. In a previous study we showed that patients' perceived loss of control and entrapment by psychotic illness (e.g. by recurring relapse) was strongly linked to depression. DESIGN: In this study we follow up the original sample of 49 patients 2.5 years later to examine the hypotheses using more powerful prospective methodology. Two of the sample had died of natural causes and the remaining 47 agreed to be reinterviewed. METHOD: We used the same measures of patients' appraisal of their illness and symptoms in terms of the extent to which they perceive it as embodying loss, humiliation and entrapment. RESULTS: It was found using multivariate analysis that perceived loss of autonomy and social role, particularly employment, were correlated with depression. The appraisal of entrapment in psychotic illness was found to have high cross sectional and prospective predictive value independent of illness, symptom and treatment variables and was shown to be influenced by certain aspects of psychiatric treatment, particularly compulsory detention. CONCLUSION: We propose that episodes of depression in schizophrenia are triggered by psychosis-related events (relapse, compulsory admission, residual voices, loss of job, etc.) that signify the inability to overcome the loss of a cherished personal goal or social role and thereby to affirm an identity. Implications for psychological therapy are discussed. PMID- 9874589 TI - [Liudmila Antoninova Shangina (on the centenary of her birth)]. PMID- 9874590 TI - Aspirin dose in secondary prevention of stroke. PMID- 9874592 TI - Immunoendocrine Interactions: A Comparative View. Proceedings of a workshop. Mariensee, Germany. PMID- 9874591 TI - Spontaneous decrease of MCA-stenosis due to cerebral thromboembolism. PMID- 9874593 TI - [Evaluation of preoperative complementary examination ordering]. AB - The ordering of tests is often done in a systematical way. In spite of the studies which have proved that these tests are not useful, and despite the advice of the SFAR (1992) the ordering remains excessive, and is a source of expenses for the population. In that study we have done an evaluation for the ordering of preoperative tests in our structure and we have tried to see if the advice of the SFAR were followed, if they were sufficient to modify the habits of ordering the tests. PMID- 9874595 TI - [Ambulatory 24-hour blood pressure monitoring]. PMID- 9874594 TI - [Tuberculosis among hemodialysis patients in Dakar, apropos of 2 cases]. AB - Hemodialysis permits a long term survival to patients with End Stage Renal Disease (E.S.R.D.). However the patients ongoing hemodialysis presented a immunodeficiency and a important modification of drugs biodisponibility. Tuberculosis is an endemic disease in our countries. Extrapulmonary tuberculosis is reported from these two cases among 13 patients treated in C.H.U.A. Le Dantec hemodialysis center. Clinical symptoms are not specific bacteria is not found. Diagnosis is obtained by a bundle of arguments. Antituberculosis treatment need to be adjusted in this field. Even if Rifampicine can be administrated at normal dose. The others drugs must be adapted on their clearance and the underlying disease. The two patients presented psychiatrical symptoms motivating a reduction of isoniazide dose witch threshold toxicity is lowered by renal failure state. These observations must increase watchfulness on nephrologist of undeveloping countries, confronted with tuberculosis renewed out break. PMID- 9874596 TI - 4th Workshop on the Neurobiology of Epilepsy (WONOEP IV). Adare, Ireland, 26-28 June 1997. PMID- 9874597 TI - Repressor elements in the coding region of the human histone H4 gene interact with the transcription factor CDP/cut. AB - The coding region of the human histone H4 gene FO108 undergoes dynamic changes in chromatin structure that correlate with modifications in gene expression. Such structural alterations generally reflect transcription factor interactions with gene regulatory sequences. To test for regulatory elements within the coding region, we performed transient transfection experiments in HeLa cells using constructs with histone H4 sequences fused upstream of a heterologous thymidine kinase promoter and CAT reporter gene. H4 gene sequences from -10 to +210 repressed transcription 4.8-fold. Further deletion and mutational analysis delineated three repressor elements within this region. Using oligonucleotide competition analysis and specific antibody recognition in electrophoretic mobility shift assays, as well as methylation interference and DNase I footprinting analyses, we have identified the CCAAT displacement protein (CDP/cut) as the factor that interacts with these three repressor elements. CDP/cut binding to these repressor sites is proliferation-specific and cell-cycle regulated, increasing in mid to late S phase. Our results indicate that the proximal 200 nucleotides of the histone H4-coding region contain transcriptional regulatory elements that may contribute to cell-cycle control of histone gene expression by interacting with repressor complexes containing CDP/cut homeodomain transcription factors. PMID- 9874598 TI - Reflux in the mouse model of urinary tract infection. PMID- 9874599 TI - Statistics on survival from Osaka Cancer Registry. PMID- 9874600 TI - Is the pulmonary pressure-volume curve symmetrical with respect to the inflection point? PMID- 9874601 TI - [Informing the patient. A threat for the surgeon or bouncing opportunity?]. PMID- 9874602 TI - Frontiers in Clinical Dentistry: Caries and Periodontal Disease. Symposium proceedings. Seattle, Washington, USA. May 21-22, 1998. PMID- 9874603 TI - Proceedings of the 3rd International Conference on the Varicella-Zoster Virus. Palm Beach Gardens, Florida, USA. 9-11 March 1997. PMID- 9874604 TI - The dawning of the age of accountability. PMID- 9874605 TI - State adopts new prescription drug rules. PMID- 9874606 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 39-1998. A 13-year-old girl with a relapsing-remitting neurologic disorder. PMID- 9874607 TI - Clodronate in metastatic breast cancer. PMID- 9874608 TI - Clodronate in metastatic breast cancer. PMID- 9874609 TI - Clodronate in metastatic breast cancer. PMID- 9874610 TI - Perinatal transmission of human granulocytic ehrlichiosis. PMID- 9874611 TI - Perinatal transmission of human granulocytic ehrlichiosis. PMID- 9874612 TI - Gains in life expectancy from medical interventions. PMID- 9874613 TI - Gains in life expectancy from medical interventions. PMID- 9874614 TI - Loxoscelism. PMID- 9874615 TI - Loxoscelism. PMID- 9874616 TI - Loxoscelism. PMID- 9874617 TI - Mastic gum kills Helicobacter pylori. PMID- 9874618 TI - Relation between chubby cheeks and visceral fat. PMID- 9874619 TI - A risk of mineral oil. PMID- 9874620 TI - The risk-adjustment debate. PMID- 9874621 TI - Two comments on Dr. Ebel's review on congenital obstruction of the urinary tract. PMID- 9874622 TI - Two comments on Dr. Ebel's review on congenital obstruction of the urinary tract. PMID- 9874623 TI - On World AIDS Day, a shadow looms over southern Africa. PMID- 9874624 TI - Kick-starting the AIDS vaccine effort. PMID- 9874625 TI - Steadying influence for neurons identified. PMID- 9874626 TI - Heat shock protein mutes genetic changes. PMID- 9874627 TI - Drug may suppress the craving for nicotine. PMID- 9874628 TI - AIDS research. New czar aims to sharpen France's effort. PMID- 9874629 TI - Geologists take a trip to the red planet. PMID- 9874630 TI - Freedom of information requests. PMID- 9874631 TI - Freedom of information requests. PMID- 9874632 TI - Human cloning. PMID- 9874633 TI - Lead regulation. PMID- 9874634 TI - Just the facts of chromatin transcription. PMID- 9874635 TI - The enigmas of Kaposi's sarcoma. PMID- 9874636 TI - Biological hydrogen production: not so elementary. PMID- 9874638 TI - Oxygen isotope exchange between refractory inclusion in Allende and solar nebula gas. AB - A calcium-aluminum-rich inclusion (CAI) from the Allende meteorite was analyzed and found to contain melilite crystals with extreme oxygen-isotope composition (approximately 5 percent oxygen-16 enrichment relative to terrestrial oxygen-16). Some of the melilite is also anomalously enriched in oxygen-16 compared with oxygen isotopes measured in other CAIs. The oxygen isotopic variation measured among the minerals (melilite, spinel, and fassaite) indicates that crystallization of the CAI started from oxygen-16-rich materials that were probably liquid droplets in the solar nebula, and oxygen isotope exchange with the surrounding oxygen-16-poor nebular gas progressed through the crystallization of the CAI. Additional oxygen isotope exchange also occurred during subsequent reheating events in the solar nebula. PMID- 9874637 TI - The rotary enzyme of the cell: the rotation of F1-ATPase. PMID- 9874639 TI - Use of stem cells still legally murky, but hearing offers hope. PMID- 9874640 TI - Scientific panel clears breast implants. PMID- 9874641 TI - Animal experimentation. India backs off on central control. PMID- 9874642 TI - Pediatric vaccines. Gates launches $100 million initiative. PMID- 9874643 TI - Worming secrets from the C. elegans genome. PMID- 9874644 TI - Bid for better beef gives Japan a leg up on cattle. PMID- 9874645 TI - Staying afloat on the seas of data. PMID- 9874646 TI - Neandertals: not so fast. PMID- 9874647 TI - The context of molecular data. PMID- 9874648 TI - Icelandic health records. PMID- 9874649 TI - Protein data bank deposits. PMID- 9874650 TI - The handy-dandy kitchen device. PMID- 9874651 TI - Proprioception and the McGurk effect. PMID- 9874652 TI - A welcome mat for leprosy and Lassa fever. PMID- 9874653 TI - Nuclear functions charge ahead. PMID- 9874654 TI - [Catalog of recent video tapes in Annali Italiani di Chirurgia and stored in the editor's office]. PMID- 9874655 TI - Differentiation between Mycobacterium bovis BCG-vaccinated and M. bovis-infected cattle by using recombinant mycobacterial antigens. AB - Tuberculosis continues to be a worldwide problem for both humans and animals. The development of tests to differentiate between infection with Mycobacterium tuberculosis or Mycobacterium bovis and vaccination with M. bovis BCG could greatly assist in the diagnosis of early infection as well as enhance the use of tuberculosis vaccines on a wider scale. Recombinant forms of four major secreted proteins of M. bovis-MPB59, MPB64, MPB70, and ESAT-6-were tested in a whole-blood gamma interferon (IFN-gamma) assay for differentiation between cattle vaccinated with BCG and those experimentally infected with M. bovis. BCG vaccination induced minimal protection in the present study, with similar numbers of animals infected with M. bovis in BCG-vaccinated and nonvaccinated groups. Following vaccination with BCG, the animals produced moderate IFN-gamma responses to bovine purified protein derivative (PPDB) but very weak responses to the recombinant antigens. Cattle from both the BCG-vaccinated and nonvaccinated groups which were M. bovis culture positive following challenge produced IFN-gamma responses to PPDB and ESAT-6 which were significantly stronger than those observed in the corresponding M. bovis culture-negative animals. IFN-gamma responses to MPB59, MPB64, and MPB70 were significantly weaker, and these antigens could not discriminate between vaccinated animals which develop disease and the culture-negative animals. The results of the study indicate that of the four antigens tested in the IFN-gamma assay, only ESAT-6 would be suitable for differentiating BCG-vaccinated animals from those infected with bovine tuberculosis. PMID- 9874656 TI - Changes in immune parameters seen in Gulf War veterans but not in civilians with chronic fatigue syndrome. AB - The purpose of this study was to evaluate immune function through the assessment of lymphocyte subpopulations (total T cells, major histocompatibility complex [MHC] I- and II-restricted T cells, B cells, NK cells, MHC II-restricted T-cell derived naive and memory cells, and several MHC I-restricted T-cell activation markers) and the measurement of cytokine gene expression (interleukin 2 [IL-2], IL-4, IL-6, IL-10, IL-12, gamma interferon [IFN-gamma], and tumor necrosis factor alpha [TNF-alpha]) from peripheral blood lymphocytes. Subjects included two groups of patients meeting published case definitions for chronic fatigue syndrome (CFS)-a group of veterans who developed their illness following their return home from participating in the Gulf War and a group of nonveterans who developed the illness sporadically. Case control comparison groups were comprised of healthy Gulf War veterans and nonveterans, respectively. We found no significant difference for any of the immune variables in the nonveteran population. In contrast, veterans with CFS had significantly more total T cells and MHC II+ T cells and a significantly higher percentage of these lymphocyte subpopulations, as well as a significantly lower percentage of NK cells, than the respective controls. In addition, veterans with CFS had significantly higher levels of IL-2, IL-10, IFN-gamma, and TNF-alpha than the controls. These data do not support the hypothesis of immune dysfunction in the genesis of CFS for sporadic cases of CFS but do suggest that service in the Persian Gulf is associated with an altered immune status in veterans who returned with severe fatiguing illness. PMID- 9874657 TI - Viability and recovery of peripheral blood mononuclear cells cryopreserved for up to 12 years in a multicenter study. AB - The Multicenter AIDS Cohort Study (MACS), an ongoing prospective study of the natural history of human immunodeficiency virus (HIV), has stored biologic specimens, including peripheral blood mononuclear cells (PBMC), from 5,622 participants for up to 12 years. The purpose of the present analysis was to evaluate the quality of the PBMC in the MACS repository in order to test the validity and feasibility of nested retrospective studies and to guide the planning of future repositories. PBMC were collected from MACS participants at four centers at 6-month intervals from 1984 to 1995, cryopreserved, and transported to a central repository for storage. A total of 596 of these specimens were subsequently tested for viability and used to evaluate cell function, to conduct immunophenotype analysis, or to isolate HIV. Simple linear regression models were applied to evaluate trends in recovery and viability over time and by center. Results indicated that from a nominal 10(7) cells cryopreserved per vial at all four centers, the median number of viable cells recovered was at least 5 x 10(6) (50% of the number stored) and the median viability was at least 90%. Results suggested that cryopreserved cells can be stored for at least 12 years with no general tendency toward cell loss over time. Furthermore, there were no statistically significant changes in the percent cell viability according to the length of time frozen, regardless of HIV serostatus or the level of CD4(+) lymphocytes. Storing 10(7) PBMC per vial yields sufficient viable cells for phenotypic and/or functional analysis. Results from the MACS provide the basis for the planning of future repositories for use by investigators with similar research goals. PMID- 9874658 TI - Evaluation of a western blot test in an outbreak of acute pulmonary histoplasmosis. AB - A western blot (WB) test was evaluated for detection of antibodies against native glycosylated and chemically deglycosylated M and H antigens of Histoplasma capsulatum in serum obtained from patients during the acute phase of pulmonary histoplasmosis that occurred during an outbreak. Of 275 serum samples tested by immunodiffusion and complement fixation (CF) samples from 40 patients affected during this outbreak and from 37 negative controls were tested by WB test. A group of patients whose sera were negative for CF antibodies and precipitins early in the acute stage of histoplasmosis but who all seroconverted during convalescence 6 weeks later were tested with the WB test. Antibodies against untreated H and M antigens were detected at a 1:100 dilution by WB test in 45% of the 20 acute-phase serum samples and in all 20 of the convalescent-phase specimens. The WB test's sensitivity for acute-phase specimens increased to 90% (18 of 20 specimens) when H and M antigens were treated by periodate oxidation to inactivate susceptible carbohydrate epitopes. When native glycosylated antigens were used in the WB test, positive reactions were observed in negative control serum specimens (3 of 37 specimens; 8%) and in serum specimens obtained from asymptomatic persons screened as part of the outbreak investigation (13 of 20 specimens; 65%). These positive reactions were also attributed to glycosidic epitopes since the specificity of the WB test increased from 78 to 100% when periodate-treated H and M antigens were used. WB test with deglycosylated H and M antigens of histoplasmin provides a rapid, sensitive, and specific test to diagnose acute pulmonary histoplasmosis before precipitins can be detected. PMID- 9874659 TI - Evaluation of recombinant dense granule antigen 7 (GRA7) of Toxoplasma gondii for detection of immunoglobulin G antibodies and analysis of a major antigenic domain. AB - Dense granule protein 7 (GRA7) of Toxoplasma gondii was expressed in Escherichia coli as a fusion protein. The leader peptide contained a 25-amino-acid mouse tumor necrosis factor fragment and six histidyl residues. After purification by metal chelate affinity chromatography, the antigen was evaluated in an enzyme linked immunosorbent assay for detection of immunoglobulin G (IgG). For two sets of IgG-positive human serum samples, obtained from routine screening, an overall sensitivity of 81% was obtained. For chronic-phase sera, the sensitivity of detection was 79%, but chronic-phase sera with low titers were more difficult to detect (65% sensitivity for sera with immunofluorescence titer of 1/64). When GRA7 was combined with Tg34AR (rhoptry protein 2 C-terminal fragment), the sensitivity rose to 96%. For a set of acute-phase serum samples tested on GRA7, the sensitivity of detection was 94%, and high-titer IgM-positive sera were detected at an especially high rate. In contrast, when Tg34AR was used, the sensitivity was only 85% for this latter set of serum samples. Three truncated GRA7 fragments containing the same leader peptide as that of recombinant GRA7 were produced. The shortest fragment (97 N-terminal amino acids) was not reactive with human sera or with a specific anti-GRA7 monoclonal antibody, while the two larger fragments were reactive. The most important antigenic domain of GRA7 for human sera was localized between residues 97 and 146. The epitope for the specific monoclonal antibody could be further narrowed down by the use of synthetic peptides, but this epitope is not recognized by sera from T. gondii infected humans. These results indicate that GRA7 may be considered as an additional tool for studying the immune response to T. gondii. PMID- 9874660 TI - Polymeric display of immunogenic epitopes from herpes simplex virus and transmissible gastroenteritis virus surface proteins on an enteroadherent fimbria. AB - The strong immunogenicity of bacterial fimbriae results from their polymeric and proteinaceous nature, and the protective role of these immunogens in experimental or commercial vaccines is associated with their capacity to induce antiadhesive antibodies. Fimbria-mediated intestinal colonization by enteropathogens typically leads to similar antibody responses. The possibility of taking advantage of these properties was investigated by determining whether enteroadhesive fimbriae, like the 987P fimbriae of enterotoxigenic Escherichia coli, can serve as carriers for foreign antigens without losing their adhesive characteristics. Random linker insertion mutagenesis of the fasA gene encoding the major 987P subunit identified five different mutants expressing wild-type levels of fimbriation. The linker insertion sites of these mutants were used to introduce three continuous segments of viral surface glycoproteins known to be accessible to antibodies. These segments encode residues 11 to 19 or 272 to 279 of herpes simplex virus type 1 (HSV-1) glycoprotein D [gD(11-19) and gD(272-279), respectively] or residues 379 to 388 of the transmissible gastroenteritis virus (TGEV) spike protein [S(379 388)]. Studies of bacteria expressing fimbriae incorporating mutated FasA subunits alone or together with wild-type FasA subunits (hybrid fimbriae) indicated that foreign epitopes were best exported and displayed on assembled fimbriae when they were inserted near the amino terminus of FasA. Fimbriated bacteria expressing FasA subunits carrying the HSV gD(11-19) or the TGEV S(379 388) epitope inserted between the second and third residues of mature FasA elicited high levels of foreign epitope antibodies in all rabbits immunized parenterally. Antibodies against the HSV epitope were also shown to recognize the epitope in the context of the whole gD protein. Because the 987P adhesive subunit FasG was shown to be present on mutated fimbriae and to mediate bacterial attachment to porcine intestinal receptors, polymeric display of foreign epitopes on 987P offers new opportunities to test the potential beneficial effect of enteroadhesion for mucosal immunization and protection against various enteric pathogens. PMID- 9874661 TI - Prevalence of Bartonella henselae and Bartonella clarridgeiae in an urban Indonesian cat population. AB - We studied evidence of Bartonella henselae and Bartonella clarridgeiae infection in 54 cats living in Jakarta, Indonesia. By using an indirect immunofluorescence assay, we found immunoglobulin G antibody to B. henselae in 40 of 74 cats (54%). The blood of 14 feral cats was cultured on rabbit blood agar plates for 28 days. Bartonella-like colonies were identified as B. henselae or B. clarridgeiae by using restriction fragment length polymorphism analysis and direct sequencing of the PCR amplicons. Of the cats sampled in the study, 6 of 14 (43%; all feral) were culture positive for B. henselae; 3 of 14 (21%; 2 feral and 1 pet) culture positive for B. clarridgeiae. This is the first report that documents B. henselae and B. clarridgeiae infections in Indonesian cats. PMID- 9874662 TI - Humoral immune response to human cytomegalovirus in patients undergoing percutaneous transluminal coronary angioplasty. AB - Possible causal relations between prior human cytomegalovirus (HCMV) infection and atherosclerosis and between HCMV reactivation and restenosis after coronary angioplasty have been suggested. We investigated patterns of antibodies directed to HCMV in 112 patients undergoing percutaneous transluminal coronary angioplasty (PTCA) and in a group of sex- and age-matched controls (blood donors without evidence of atherosclerosis). Levels of antibodies to HCMV were measured by enzyme-linked immunosorbent assay (ELISA) of serum samples drawn before and 5 weeks after PTCA. To further differentiate the humoral immune response, we specifically tested antibody reactivity towards four single HCMV proteins (IE2, p52, pp150, and pp65) by recombinant ELISAs. We found that 73% of PTCA patients and 69% of sex- and age-matched controls were seropositive for HCMV (odds ratio, 1.2 [not significant]). The corresponding odds ratios for matched pairs ranged in the recombinant ELISAs from 1.2 to 1.4. Patients had more often high titers of anti-HCMV antibodies (11 versus 4%; odds ratio = 3.3 [0.9 to 15.2]; P = 0.052) and high titers of anti-pp150 antibodies (13 versus 4%; odds ratio = 6.0 [1.3 to 38.8]; P = 0.008). Anti-HCMV immunoglobulin M antibodies were not detected in any patient. There was no evidence of acute HCMV reactivation after PTCA, since the titers of antibodies to the investigated recombinant proteins did not increase at 5 weeks after PTCA. Our results show a limited association between prior HCMV infection and coronary artery disease. We infer that positive anti-HCMV titers are not a major risk factor at the time of disease manifestation. However, this study cannot rule out a possible role of HCMV at earlier stages of the atherosclerotic process. Recombinant ELISAs provide a valuable tool for investigating the antiviral immune response. PMID- 9874664 TI - Humoral response in Toscana virus acute neurologic disease investigated by viral protein-specific immunoassays. AB - The Toscana virus (family Bunyaviridae, genus Phlebovirus) is the only sandfly transmitted virus that demonstrates neurotropic activity. Clinical cases ranging from aseptic meningitis to meningoencephalitis caused by Toscana virus are yearly observed in central Italy during the summer, and several cases have been reported among tourists returning from zones of endemicity (Italy, Portugal, Spain, and Cyprus). In Toscana virus patients, immunoglobulin M (IgM) antibodies, usually present at the onset of symptoms, can reveal elevated titers by enzyme-linked immunosorbent assay and can persist for at least 1 year. IgG antibodies can be absent at the onset of symptoms: titers rise in convalescent sera and persist for many years. At least five proteins have been identified in Toscana virus-infected cells: nucleoprotein N, glycoproteins G1 and G2, a large protein (L) assumed to be a component of the polymerase, and two nonstructural proteins, NSm and NSs. We report results of a study on the antibody response to individual viral proteins in patients with Toscana virus-associated acute neurologic disease. Immunoblotting and semiquantitative radioimmunoprecipitation assay (RIPA) allow identification of nucleoprotein N as the major antigen responsible for both IgM and IgG responses. Antibodies to proteins other than nucleoprotein N are detected only by RIPA. Antibodies to glycoproteins are detected in about one-third of patients, and whereas their presence always predicts neutralization, some serum samples with neutralizing activity have undetectable levels of antibodies to G1 G2. Antibodies to nonstructural proteins NSm and NSs are also identified. The results obtained raise some questions about antigenic variability and relevant neutralization epitopes of Toscana virus. PMID- 9874665 TI - The cured immune phenotype achieved by treatment of visceral leishmaniasis in the BALB/c mouse with a nonionic surfactant vesicular formulation of sodium stibogluconate does not protect against reinfection. AB - Single-dose treatment with sodium stibogluconate solution (SSG) and treatment with a nonionic surfactant vesicular formulation of sodium stibogluconate (SSG NIV) were compared for the ability to protect BALB/c mice against infection with Leishmania donovani. Prophylactic treatment with SSG-NIV protected against infection, although its effects were time and organ dependent; protection was not obtained with SSG. Protection against reinfection with L. donovani was observed only in mice cured by treatment with SSG-NIV. However, this protective effect was probably due to the presence of residual drug rather than an immune effect, since prophylactic SSG-NIV treatment gave similar results. Transfer of enriched spleen T-cell populations from L. donovani-infected mice or from infected SSG-NIV treated mice gave no protection against L. donovani infection in the recipients. T cells from infected mice, but not from infected SSG-NIV-treated mice, were infectious to recipients. SSG-NIV treatment was equally effective against visceral leishmaniasis in immunocompetent and SCID mice, whereas SSG treatment was less effective in the latter. The results of this study suggest that the high antileishmanial activity of SSG-NIV is due to favorable modification of SSG delivery and does not require a fully functional immune response. Cure of visceral leishmaniasis by SSG-NIV treatment in the BALB/c mouse did not protect against reinfection. PMID- 9874663 TI - Transplacental transmission of serotype-specific pneumococcal antibodies in a Brazilian population. AB - The highest incidence of severe pneumococcal infections in children occurs in the first 6 months of life; however, immunization of infants with the existing polysaccharide vaccines is ineffective. We wished to determine the prevalence of immunoglobulin G (IgG) pneumococcal antibodies in unimmunized Brazilian mothers and their transplacental transmission to term and preterm infants. Total IgG, IgG1 and -2 subclass levels, and IgG antibodies against Streptococcus pneumoniae serotypes 1, 3, 6B, 9V, and 14 were determined in 15 pairs of mothers and term newborns (gestational age, >/=37 weeks) and in 18 pairs of mothers and preterm newborns (gestational age, 32 to 36 weeks). Serotype-specific anti-pneumococcal antibodies were detected by a recently standardized enzyme-linked immunosorbent assay calibrated with the 89-SF reference serum. Varying percentages of the mothers had antibody concentrations below arbitrarily defined protective levels: 33% for serotype 1, 67% for serotype 3, 30% for serotype 6B, 52% for serotype 9V, and 22% for serotype 14. In term newborns, IgG1 concentrations were slightly higher than maternal concentrations; in preterm newborns, the concentrations were much lower. Concentrations of IgG2 in term and preterm infants were significantly lower than in the mothers. Transplacental transmission of antibodies to serotypes 3 and 14 was clearly different from that of antibodies to serotypes 1, 6B, and 9V. Concentrations of IgG antibodies against serotypes 3 and 14 were similar to or higher than those of the mothers; against serotypes 1, 6B, and 9V they ranged from 77 to 83% of maternal concentrations in term newborns and also in preterm infants, although transplacental transmission of antibodies was proportionally lower for each specific serotype in preterm than in term infants. These data are relevant for developing strategies to protect infants against pneumococcal infections in the first months of life. Our findings and a review of existing information stress the importance of understanding the relationships among pneumococcal immunization, IgG subclass antibodies to individual serotypes, transplacental transport, half-life, and antibody function and their protective values against infection. PMID- 9874666 TI - Validation of the indirect MAP1-B enzyme-linked immunosorbent assay for diagnosis of experimental Cowdria ruminantium infection in small ruminants. AB - The major antigenic protein 1 fragment B (MAP1-B) enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Cowdria ruminantium infections was validated to determine cutoff values and evaluate its diagnostic performance with sheep and goat sera. Cowdria-infected populations consisted of 48 sheep and 44 goats, while the noninfected populations consisted of 64 sheep and 107 goats. Cutoff values were determined by two-graph receiver-operating characteristic (TG-ROC) curves. The cutoff value was set at 31 and 26.6% of the positive control reference samples for sheep and goat sera, respectively. The test's diagnostic performance was evaluated with measurements of the area under the concentration-time curve (AUC) of the ROC curves and by the valid range proportion (VRP). The AUCs were 0.978 for sheep sera and 0.989 for goat sera. The VRP for both sheep and goat sera was approximately 1.0. The intermediate range (IR), which defines results that are neither positive nor negative, was 0 for goat sera and 2.81 for sheep sera. In an ideal test, the AUC and VRP would be 1.0 and the IR would be 0. In this study these parameters were close to those of an ideal test. It is concluded that the MAP1-B ELISA is a useful test for the diagnosis of C. ruminantium infection in small ruminants. PMID- 9874667 TI - Immune determinants of organism and outcome in febrile hospitalized Thai patients with bloodstream infections. AB - Opportunistic infections (OI) and the human immunodeficiency virus (HIV) cause significant morbidity and mortality in developing countries. Immune cell and cytokine profiles may be related to the type and course of OI and to the OI-HIV interaction. Examining cell-specific cytokine production ex vivo has only recently become feasible. In Thailand, 53 febrile, hospitalized adults were enrolled in a study of the immune correlates of bloodstream infections (BSI). On site, blood cells were stimulated ex vivo. Cell-surface antigens and eight intracellular cytokines were subsequently analyzed using flow cytometry to determine associations with mortality and the organism causing the BSI. By logistic regression analysis, the percentage of CD3(+) CD16/56(+) cells making tumor necrosis factor alpha (TNF-alpha) (P = 0.033) and the percentage of CD3(-) CD16/56(+) cells (NK) (P = 0.032) were related to HIV positivity. Lymph node enlargement with HIV infection and the percentage of CD3(+) CD16/56(+) making TNF alpha were predictive of death. A lower percentage of CD3(+) CD8(+) lymphocytes making interleukin-8 (IL-8) (P = 0.005), fewer monocytes expressing CD14 (P = 0.009), and the percentage of CD3(+) CD8(+) cells producing gamma interferon (P = 0. 011) were associated with blood culture positivity and the causative organism. For every one point decrease in the percentage of CD3(+) CD8(+) cells making IL 8, the likelihood of a positive culture increased 23%; for every one point decrease in the percentage of monocytes expressing CD14, the likelihood of a positive culture increased by 5%. Only a few immune cell types and three of their related cytokines were significantly associated with HIV disease outcome or the BSI organism. These cell types did not include CD3(+) CD8(-) cells (a surrogate for CD4(+) cells), nor did they involve cytokines associated with a type I to type II cytokine shift, which might occur with advancing HIV infection. These associations support the premise that CD8(+) and CD16/56(+) lymphocytes play significant roles in HIV and type I infections. PMID- 9874668 TI - Dysregulated synthesis of intracellular type 1 and type 2 cytokines by T cells of patients with cutaneous T-cell lymphoma. AB - Mycosis fungoides (MF) and Sezary syndrome (SS) are the two main clinical entities of cutaneous T-cell lymphoma (CTCL). As the disease progresses from MF to SS, a switch from a type 1 (interleukin [IL]-2 and gamma interferon [IFN gamma]) to a type 2 (IL-4) cytokine production profile occurs. Although roles for type 1 and type 2 cytokines in the pathogenesis of CTCL have been proposed, the cellular origins of these cytokines are unclear. Using flow cytometry to identify individual T-cell subsets, we studied cytokine synthesis by the T cells of 13 patients with SS and 12 with MF and 9 hematologically healthy donors. Upon activation with phorbol 12-myristate 13-acetate (PMA), the numbers of T cells synthesizing IL-2 were similar for all study groups. Whereas the predominant T cell producing IL-2 in healthy donors and in those with MF was CD7(+), in patients with SS, it was CD7(-). Although the number of IL-4(+) CD4(+) T cells was low for all study groups, there was a significantly higher number of IL-4(+) CD8(+) T cells in patients with MF than in those with SS or healthy donors. There was a decline in the number of IFN-gamma-producing T cells in CTCL donors compared to that in healthy donors. More importantly, there was a significant decrease in the number of IFN-gamma-producing T cells with disease progression from MF to SS. The inability of these T cells to synthesize IFN-gamma may have prognostic value in CTCL, since it may be responsible for the progression of the disease from MF to SS. PMID- 9874669 TI - Quantitative analysis of T-cell receptor beta variable-gene usage in cutaneous late-phase reactions: implications for T-lymphocyte recruitment in cutaneous inflammation. AB - To determine if functionally distinct T-lymphocyte (T cell) subsets accumulate in late-phase immunoglobulin E-mediated reactions (LPR), we quantitatively analyzed the immunophenotype and the T-cell receptor beta variable-gene (Vbeta) repertoire of T cells in cutaneous LPR. Peripheral blood and skin biopsies were obtained 6 or 24 h after sensitive subjects were challenged with intradermal injections of grass pollen allergen (Ag) and control (C) solution. The frequency of cells expressing CD3, CD4, CD8, CD45RO, and CD25/mm2 was determined by immunohistochemistry in nine subjects. Vbeta usage was assessed by reverse transcription-PCR in five of nine subjects. A significantly greater frequency of CD3(+) and CD45RO+ (memory) T cells was detected in Ag sites than in C sites at 24 h after challenge but not at 6 h. The frequency of activated (CD25(+)) and helper (CD4(+)) T cells appeared to be increased in Ag sites as well, though not significantly. Vbeta6 was the most commonly expressed Vbeta detected in Ag sites, but it was also detected in accompanying C sites. Vbeta2 was the most commonly expressed Vbeta detected in C sites. Sequence analysis in one case revealed Vbeta expression in a 6-h Ag site to be essentially polyclonal. Our findings suggest that memory T cells with Vbeta expression similar to that in normal skin accumulate in developing cutaneous LPR. The limited usage of Vbeta suggests a preferential recruitment or retention of reactive T cells from an endogenous subset of skin-homing T cells with its own skewed Vbeta repertoire. PMID- 9874671 TI - Identification of common lipooligosaccharide types in isolates from patients with otitis media by monoclonal antibodies against nontypeable Haemophilus influenzae 9274. AB - Twenty-one murine monoclonal antibodies (MAbs) were induced by nontypeable Haemophilus influenzae (NTHi) 9274. Nineteen MAbs were specific for the lipooligosaccharide (LOS) as determined by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. When the MAbs were assayed with five LOS prototype strains by ELISA, all bound to strain 3198 LOS (type III), while six of the MAbs were also reactive with LOSs from strain 1479 (type I), 5657 (type IV), or 7502 (type V). Ten MAbs had complement-mediated bactericidal activity, and three MAbs were opsonophagocytic against the homologous strain. Five LOS MAbs with different specificities were used to analyze 155 NTHi clinical isolates from the United States and from Japan. These isolates were classified into nine groups by ELISA. Only four isolates (2.6%) were not recognized by any of the five MAbs. Most of the isolates (91.6%) were in four groups which bound three of the five MAbs. One of three MAbs, 6347C11, had strong activity against the homologous strain and was also bactericidal to 45 clinical isolates (29%) which belonged to the four common patterns (25 belonged to pattern 1). These data indicate that these MAbs can be used for LOS typing in which almost all NTHi strains can be typed according to the LOS antigenicity. Among NTHi, at least one conserved LOS epitope which is a target of bactericidal antibodies exists. We conclude that strain 9274 LOS, which is the target for bactericidal antibodies, is a candidate for LOS-based NTHi vaccines. PMID- 9874670 TI - Variables that affect assays for plasma cytokines and soluble activation markers. AB - Cytokines and soluble immune activation markers that reflect cytokine activities in vivo are increasingly being measured in plasma, serum, and other body fluids. They provide useful diagnostic and prognostic information as well as insight into disease pathogenesis. Assays of neopterin, beta2-microglobulin, soluble interleukin-2 receptor, and soluble tumor necrosis factor receptor type II as well as of the cytokines tumor necrosis factor alpha and gamma interferon (IFN gamma) were evaluated by using serum and plasma samples of human immunodeficiency virus (HIV)-positive and HIV-negative subjects. Many factors were found to influence the outcomes of these assays. Substantial differences in apparent levels of analytes were frequently found when enzyme-linked immunosorbent assay (ELISA) kits from different manufacturers were used. In some cases, differences were found in the standards provided by separate manufacturers. Furthermore, the analytic results from different lots of ELISA kits supplied by single manufacturers differed by as much as 50%. The need for uniformity in the standards for quantitative assays was clearly illustrated. International reference standards are available for cytokines but not for soluble cytokine receptors or soluble activation markers. Marker levels in serum or in plasma were similar except those for IFN-gamma. Most of the analytes were stable under several storage conditions. Thus, batch testing of frozen stored samples is feasible. The findings indicate that for longitudinal studies, the levels of cytokines and immune activation markers in plasma or serum should be measured by using preverified reagents from one manufacturer. The quality of laboratory performance can have an impact on clinical relevance. Proficiency testing and external quality assurance programs can help to develop the needed consensus. PMID- 9874672 TI - Ceramidase activity in bacterial skin flora as a possible cause of ceramide deficiency in atopic dermatitis. AB - A marked decrease in the content of ceramide has been reported in the horny layer of the epidermis in atopic dermatitis (AD). This decrease impairs the permeability barrier of the epidermis, resulting in the characteristic dry and easily antigen-permeable skin of AD, since ceramide serves as the major water holding molecule in the extracellular space of the horny layer. On the other hand, the skin of such patients is frequently colonized by bacteria, most typically by Staphylococcus aureus, possessing genes such as those for sphingomyelinase, which are related to sphingolipid metabolism. We therefore tried to identify a possible correlation between the ceramide content and the bacterial flora obtained from the skin of 25 patients with AD versus that of 24 healthy subjects, using a thin-layer chromatographic assay of the sphingomyelin associated enzyme activities secreted from the bacteria. The findings of the assay demonstrated that ceramidase, which breaks ceramide down into sphingosine and fatty acid, was secreted significantly more from the bacterial flora obtained from both the lesional and the nonlesional skin of patients with AD than from the skin of healthy subjects; sphingomyelinase, which breaks sphingomyelin down into ceramide and phosphorylcholine, was secreted from the bacterial flora obtained from all types of skin at similar levels for the patients with AD and the healthy controls. The finding that the skin of patients with AD is colonized by ceramidase-secreting bacteria thus suggests that microorganisms are related to the deficiency of ceramide in the horny layer of the epidermis, which increases the hypersensitivity of skin in AD patients by impairing the permeability barrier. PMID- 9874673 TI - Angiocentric CD3(+) T-cell infiltrates in human immunodeficiency virus type 1 associated central nervous system disease in children. AB - A significant proportion of brain tissue specimens from children with AIDS show evidence of vascular inflammation in the form of transmural and/or perivascular mononuclear-cell infiltrates at autopsy. Previous studies have shown that in contrast to inflammatory lesions observed in human immunodeficiency virus type 1 (HIV-1) encephalitis, in which monocytes/macrophages are the prevailing mononuclear cells, these infiltrates consist mostly of lymphocytes. Perivascular mononuclear-cell infiltrates were found in brain tissue specimens collected at autopsy from five of six children with AIDS and consisted of CD3(+) T cells and equal or greater proportions of CD68(+) monocytes/macrophages. Transmural (including endothelial) mononuclear-cell infiltrates were evident in one patient and comprised predominantly CD3(+) T cells and small or, in certain vessels, approximately equal proportions of CD68(+) monocytes/macrophages. There was a clear preponderance of CD3(+) CD8(+) T cells on the endothelial side of transmural infiltrates. In active lesions of transmural vasculitis, CD3(+) T-cell infiltrates exhibited a distinctive zonal distribution. The majority of CD3(+) cells were also CD8(+) and CD45RO+. Scattered perivascular monocytes/macrophages in foci of florid vasculitis were immunoreactive for the p24 core protein. In contrast to the perivascular space, the intervening brain neuropil was dominated by monocytes/macrophages, microglia, and reactive astrocytes, containing only scant CD3(+) CD8(+) cells. Five of six patients showed evidence of calcific vasculopathy, but only two exhibited HIV-1 encephalitis. One patient had multiple subacute cerebral and brainstem infarcts associated with a widespread, fulminant mononuclear-cell vasculitis. A second patient had an old brain infarct associated with fibrointimal thickening of large leptomeningeal vessels. These infiltrating CD3(+) T cells may be responsible for HIV-1-associated CNS vasculitis and vasculopathy and for endothelial-cell injury and the opening of the blood-brain barrier in children with AIDS. PMID- 9874675 TI - Anticytomegalovirus (anti-CMV) immunoglobulin G avidity in identification of pregnant women at risk of transmitting congenital CMV infection. AB - In this work, we show that the determination of the anticytomegalovirus antibody avidity carried out before week 18 of gestation is a helpful tool to identify women for enrollment in prenatal diagnosis. This procedure can identify all pregnant women who will give birth to an infected newborn. PMID- 9874674 TI - Giant syncytia and virus-like particles in ovarian carcinoma cells isolated from ascites fluid. AB - Ovarian cancer cells were isolated from ascites fluid of 30 different patients diagnosed with cystadenocarcinoma of ovaries. Large colonies of malignant ASC cells were observed during the first week of cell growth in vitro. Colony formation was followed by fusion of cells and formation of large multinucleated and highly vacuolated syncytia. In contrast, cells isolated from the ascites fluid produced by patients with benign mucinous cystadenoma of ovaries did not form syncytia. Nonmalignant Brenner tumor cells, isolated from the ascites fluid, also did not form syncytia. Syncytia, but not the nonmalignant tumor cells, were immunofluorescence stained with an anti-human immunodeficiency virus type 1 (HIV 1) gp120 monoclonal antibody (MAb) and MAb RAK-BrI. Both MAbs recognized cancer associated antigens RAK (for Rakowicz markers) p120, p42, and p25. Exposure of ASC cells to either the anti-HIV-1 gp120 MAb or MAb RAK-BrI inhibited syncytium formation. PCR with HIV-1 Env-derived primers revealed DNA sequences with over 90% homology to HIV-1 gp41 in syncytia and in ovarian cancer cells but not in normal ovary cells. Electron microscopic analysis revealed viral particles, hexagonal in shape (90 nm in diameter), with a dense central core surrounded by an inner translucent capsid and dense outer shell with projections. Negative staining detected membrane-covered particles (100 to 110 nm in diameter) in the cell culture medium. Incubation of normal breast cells with viral particles resulted in drastic morphological changes and syncytium formation by the transformed breast cells. The cytopathic effects of the identified virus resembled those of spumaviruses, which, in addition to their epitopic and genetic homology to HIV-1, might suggest a common phylogeny. PMID- 9874676 TI - Development and applications of a bovine coronavirus antigen detection enzyme linked immunosorbent assay. AB - We developed a monoclonal antibody-based, antigen capture sandwich enzyme-linked immunosorbent assay (ELISA) for bovine coronavirus. We compared the ELISA with electron microscopy and the hemagglutination test and found a close correlation between them. The sensitivity of the ELISA was 10(4) bovine coronavirus particles per ml of 10% fecal suspension. Compared with electron microscopy, bovine coronavirus ELISA had 96% specificity. PMID- 9874677 TI - Streptococcal DNase B is immunologically identical to superantigen SpeF but involves separate domains. AB - The previous suggestion that streptococcal superantigen SpeF might be identical to DNase B was confirmed in this study. Polyclonal SpeF-specific antisera were able to inhibit depolymerization of methyl-green DNA by DNase B. However, T-cell mitogenicity and nuclease activity appear to involve separate immune epitopes on SpeF, since sera with the capacity to neutralize the mitogenic activity of SpeF did not always inhibit the DNase activity. PMID- 9874678 TI - Comparison of three different methods for measuring classical pathway complement activity. AB - The complement system plays an important role in host defense against infection and in most inflammatory processes. The standard 50% hemolytic complement (CH50) assay is the most commonly used method of screening patient sera for functional activity of the classical complement pathway. Our objective in this study was to compare two newer methods (the enzyme immunoassay and the liposome immunoassay) to a commercial CH50 assay for measuring total classical complement activity. We conclude that both newer methods compare well with a CH50 assay and are equally sensitive in screening routine clinical sera. PMID- 9874679 TI - Incidence of human immunodeficiency virus antibody in a prenatal population at a community hospital. AB - Prenatal human immunodeficiency virus (HIV) screening may reduce vertical HIV transmission. We screened 4,419 prenatal sera and found 38 repeatedly reactive specimens with an HIV-1-HIV-2 enzyme-linked immunosorbent assay. Western blot analysis confirmed four of these specimens as positive for HIV-1 antibodies. Screening detects previously unidentified HIV infections, but false-positive results may also occur. PMID- 9874682 TI - Editorial PMID- 9874683 TI - Commentary: A revised view of local anesthetic action: what channel state is really stabilized? PMID- 9874680 TI - Expression and self-assembly of Grimsby virus: antigenic distinction from Norwalk and Mexico viruses. AB - A cDNA obtained from Grimsby virus (GRV), a Norwalk-like virus, purified from a stool sample of a symptomatic adult associated with a gastroenteritis outbreak in the United Kingdom, was used to obtain the complete nucleotide sequence of the second open reading frame (ORF2). The ORF2 sequence of GRV predicts a capsid of 539 amino acids (aa) which exhibits aa identities of 96% to Lordsdale virus, 67% to Mexico virus (MXV), and 43% to Norwalk virus (NV). The GRV capsid protein was expressed in insects cells by using a recombinant baculovirus, and the resulting virus-like particles (VLPs) possessed a protein with an apparent molecular weight of 58,000. Hyperimmune antisera raised against purified GRV, MXV, and NV VLPs were tested in an indirect enzyme-linked immunosorbent assay (ELISA) against GRV, NV, and MXV VLPs, revealing that GRV is antigenically distinct from both NV and MXV. The antigenic specificity of the GRV-hyperimmune antiserum was confirmed in an antigen capture ELISA using GRV-, NV-, or MXV-containing fecal specimens. The expression of the GRV capsid protein has, for the first time, allowed the antigenic comparison of three distinct recombinant Norwalk-like viruses. PMID- 9874684 TI - The position of the fast-inactivation gate during lidocaine block of voltage gated Na+ channels. AB - Lidocaine produces voltage- and use-dependent inhibition of voltage-gated Na+ channels through preferential binding to channel conformations that are normally populated at depolarized potentials and by slowing the rate of Na+ channel repriming after depolarizations. It has been proposed that the fast-inactivation mechanism plays a crucial role in these processes. However, the precise role of fast inactivation in lidocaine action has been difficult to probe because gating of drug-bound channels does not involve changes in ionic current. For that reason, we employed a conformational marker for the fast-inactivation gate, the reactivity of a cysteine substituted at phenylalanine 1304 in the rat adult skeletal muscle sodium channel alpha subunit (rSkM1) with [2 (trimethylammonium)ethyl]methanethiosulfonate (MTS-ET), to determine the position of the fast-inactivation gate during lidocaine block. We found that lidocaine does not compete with fast-inactivation. Rather, it favors closure of the fast inactivation gate in a voltage-dependent manner, causing a hyperpolarizing shift in the voltage dependence of site 1304 accessibility that parallels a shift in the steady state availability curve measured for ionic currents. More significantly, we found that the lidocaine-induced slowing of sodium channel repriming does not result from a slowing of recovery of the fast-inactivation gate, and thus that use-dependent block does not involve an accumulation of fast inactivated channels. Based on these data, we propose a model in which transitions along the activation pathway, rather than transitions to inactivated states, play a crucial role in the mechanism of lidocaine action. PMID- 9874685 TI - C-Linker of cyclic nucleotide-gated channels controls coupling of ligand binding to channel gating. AB - Cyclic nucleotide-gated channels are composed of a core transmembrane domain, structurally homologous to the voltage-gated K+ channels, and a cytoplasmic ligand-binding domain. These two modules are joined by approximately 90 conserved amino acids, the C-linker, whose precise role in the mechanism of channel activation by cyclic nucleotides is poorly understood. We examined cyclic nucleotide-gated channels from bovine photoreceptors and Caenorhabditis elegans sensory neurons that show marked differences in cyclic nucleotide efficacy and sensitivity. By constructing chimeras from these two channels, we identified a region of 30 amino acids in the C-linker (the L2 region) as an important determinant of activation properties. An increase in both the efficacy of gating and apparent affinity for cGMP and cAMP can be conferred onto the photoreceptor channel by the replacement of its L2 region with that of the C. elegans channel. Three residues within this region largely account for this effect. Despite the profound effect of the C-linker region on ligand gating, the identity of the C linker does not affect the spontaneous, ligand-independent open probability. Based on a cyclic allosteric model of activation, we propose that the C-linker couples the opening reaction in the transmembrane core region to the enhancement of the affinity of the open channel for agonist, which underlies ligand gating. PMID- 9874686 TI - Blockade of a retinal cGMP-gated channel by polyamines. AB - The cyclic nucleotide-gated (CNG) channel in retinal rods converts the light regulated intracellular cGMP concentration to various levels of membrane potential. Blockade of the channel by cations such as Ca2+ and Mg2+ lowers its effective conductance. Consequently, the membrane potential has very low noise, which enables rods to detect light with extremely high sensitivity. Here, we report that three polyamines (putrescine, spermidine, and spermine), which exist in both the intracellular and extracellular media, also effectively block the CNG channel from both sides of the membrane. Among them, spermine has the greatest potency. Extracellular spermine blocks the channel as a permeant blocker, whereas intracellular spermine appears to block the channel in two conformations-one permeant, and the other non- (or much less) permeant. The membrane potential in rods is typically depolarized to approximately -40 mV in the dark. At this voltage, K1/2 of the CNG channel for extracellular spermine is 3 microM, which is 100-1,000-fold higher affinity than that of the NMDA receptor-channel for extracellular spermine. Blockade of the CNG channel by polyamines may play an important role in suppressing noise in the signal transduction system in rods. PMID- 9874687 TI - Noncatalytic inhibition of cyclic nucleotide-gated channels by tyrosine kinase induced by genistein. AB - Rod photoreceptor cyclic nucleotide-gated (CNG) channels are modulated by tyrosine phosphorylation. Rod CNG channels expressed in Xenopus oocytes are associated with constitutively active protein tyrosine kinases (PTKs) and protein tyrosine phosphatases that decrease and increase, respectively, the apparent affinity of the channels for cGMP. Here, we examine the effects of genistein, a competitive inhibitor of the ATP binding site, on PTKs. Like other PTK inhibitors (lavendustin A and erbstatin), cytoplasmic application of genistein prevents changes in the cGMP sensitivity that are attributable to tyrosine phosphorylation of the CNG channels. However, unlike these other inhibitors, genistein also slows the activation kinetics and reduces the maximal current through CNG channels at saturating cGMP. These effects occur in the absence of ATP, indicating that they do not involve inhibition of a phosphorylation event, but rather involve an allosteric effect of genistein on CNG channel gating. This could result from direct binding of genistein to the channel; however, the time course of inhibition is surprisingly slow (>30 s), raising the possibility that genistein exerts its effects indirectly. In support of this hypothesis, we find that ligands that selectively bind to PTKs without directly binding to the CNG channel can nonetheless decrease the effect of genistein. Thus, ATP and a nonhydrolyzable ATP derivative competitively inhibit the effect of genistein on the channel. Moreover, erbstatin, an inhibitor of PTKs, can noncompetitively inhibit the effect of genistein. Taken together, these results suggest that in addition to inhibiting tyrosine phosphorylation of the rod CNG channel catalyzed by PTKs, genistein triggers a noncatalytic interaction between the PTK and the channel that allosterically inhibits gating. PMID- 9874688 TI - A serine residue in ClC-3 links phosphorylation-dephosphorylation to chloride channel regulation by cell volume. AB - In many mammalian cells, ClC-3 volume-regulated chloride channels maintain a variety of normal cellular functions during osmotic perturbation. The molecular mechanisms of channel regulation by cell volume, however, are unknown. Since a number of recent studies point to the involvement of protein phosphorylation/dephosphorylation in the control of volume-regulated ionic transport systems, we studied the relationship between channel phosphorylation and volume regulation of ClC-3 channels using site-directed mutagenesis and patch clamp techniques. In native cardiac cells and when overexpressed in NIH/3T3 cells, ClC-3 channels were opened by cell swelling or inhibition of endogenous PKC, but closed by PKC activation, phosphatase inhibition, or elevation of intracellular Ca2+. Site-specific mutational studies indicate that a serine residue (serine51) within a consensus PKC-phosphorylation site in the intracellular amino terminus of the ClC-3 channel protein represents an important volume sensor of the channel. These results provide direct molecular and pharmacological evidence indicating that channel phosphorylation/dephosphorylation plays a crucial role in the regulation of volume sensitivity of recombinant ClC-3 channels and their native counterpart, ICl.vol. PMID- 9874689 TI - Distinct structural requirements for clustering and immobilization of K+ channels by PSD-95. AB - PDZ-domain-containing proteins such as PSD-95 have been implicated in the targeting and clustering of membrane proteins. Biochemical and immunohistochemical studies indicate that PSD-95 recognizes COOH-terminal S/TXV sequences present in Kv1 K+ channels. However, the effect of binding a PDZ domain on a target protein has not been studied in live cells. In the present study, a green fluorescent protein-Kv1.4 fusion protein is used to study the effect of PSD 95 on channel movement. Fluorescence recovery after photobleaching showed that PSD-95 can immobilize K+ channels in the plasma membrane in an all-or-none manner. Furthermore, time lapse imaging showed that channel clusters formed in the presence of PSD-95 are stable in size, shape, and position. As expected from previous reports, two green fluorescent protein-tagged COOH-terminal variants of Kv1.4, Delta15 and V655A, are not clustered by PSD-95. However, coexpression of PSD-95 with V655A, but not Delta15, leads to the appearance of PSD-95 immunoreactivity in the plasma membrane. Furthermore, fluorescence recovery after photobleaching studies show that V655A channels are immobilized by PSD-95. Thus, V655A channels can interact with PSD-95 in a manner that leads to channel immobilization, but not clustering. These experiments document for the first time that PSD-95 immobilizes target proteins. Additionally, the data presented here demonstrate that the structural requirements for protein clustering and immobilization by PSD-95 are distinct. PMID- 9874690 TI - Antisense knock out of the inositol 1,3,4,5-tetrakisphosphate receptor GAP1(IP4BP) in the human erythroleukemia cell line leads to the appearance of intermediate conductance K(Ca) channels that hyperpolarize the membrane and enhance calcium influx. AB - To study the role of the inositol 1,3,4,5-trisphosphate-binding protein GAP1(IP4BP) in store-operated Ca2+ entry, we established a human erythroleukemia (HEL) cell line in which the expression of GAP1(IP4BP) was substantially reduced by transfection with a vector containing antisense DNA under control of a Rous Sarcoma virus promoter and the Escherichia coli LacI repressor (AS-HEL cells). Control cells were transfected with vector lacking antisense DNA (V-HEL cells). GAP1(IP4BP) protein, which is a member of the GTPase-activating protein (GAP1) family, was reduced by 85% in AS-HEL cells and was further reduced by 96% by treatment with isopropylthio-beta-D- galactoside to relieve LacI repression. The loss of GAP1(IP4BP) was associated with both a membrane hyperpolarization and a substantially increased Ca2+ entry induced by thrombin or thapsigargin. The activation of intermediate conductance Ca2+-activated K+ channels in AS-HEL cells (not seen in V-HEL cells) was responsible for the membrane hyperpolarization and the enhanced Ca2+ entry, and both were blocked by charybdotoxin. Stimulated V-HEL cells did not hyperpolarize and basal Ca2+ influx was unaffected by charybdotoxin. In V-HEL cells hyperpolarized by removal of extracellular K+, the thapsigargin-stimulated Ca2+ influx was increased. Expression of mRNA for the human Ca2+-activated intermediate conductance channel KCa4 was equivalent in both AS-HEL and V-HEL cells, suggesting that the specific appearance of calcium activated potassium current (IK(Ca)) in AS-HEL cells was possibly due to modulation of preexisting channels. Our results demonstrate that GAP1(IP4BP), likely working through a signaling pathway dependent on a small GTP-binding protein, can regulate the function of K(Ca) channels that produce a hyperpolarizing current that substantially enhances the magnitude and time course of Ca2+ entry subsequent to the release of internal Ca2+ stores. PMID- 9874691 TI - Regulators of G protein signaling attenuate the G protein-mediated inhibition of N-type Ca channels. AB - Regulators of G protein signaling (RGS) proteins bind to the alpha subunits of certain heterotrimeric G proteins and greatly enhance their rate of GTP hydrolysis, thereby determining the time course of interactions among Galpha, Gbetagamma, and their effectors. Voltage-gated N-type Ca channels mediate neurosecretion, and these Ca channels are powerfully inhibited by G proteins. To determine whether RGS proteins could influence Ca channel function, we recorded the activity of N-type Ca channels coexpressed in human embryonic kidney (HEK293) cells with G protein-coupled muscarinic (m2) receptors and various RGS proteins. Coexpression of full-length RGS3T, RGS3, or RGS8 significantly attenuated the magnitude of receptor-mediated Ca channel inhibition. In control cells expressing alpha1B, alpha2, and beta3 Ca channel subunits and m2 receptors, carbachol (1 microM) inhibited whole-cell currents by approximately 80% compared with only approximately 55% inhibition in cells also expressing exogenous RGS protein. A similar effect was produced by expression of the conserved core domain of RGS8. The attenuation of Ca current inhibition resulted primarily from a shift in the steady state dose-response relationship to higher agonist concentrations, with the EC50 for carbachol inhibition being approximately 18 nM in control cells vs. approximately 150 nM in RGS-expressing cells. The kinetics of Ca channel inhibition were also modified by RGS. Thus, in cells expressing RGS3T, the decay of prepulse facilitation was slower, and recovery of Ca channels from inhibition after agonist removal was faster than in control cells. The effects of RGS proteins on Ca channel modulation can be explained by their ability to act as GTPase-accelerating proteins for some Galpha subunits. These results suggest that RGS proteins may play important roles in shaping the magnitude and kinetics of physiological events, such as neurosecretion, that involve G protein-modulated Ca channels. PMID- 9874692 TI - Gating of single N-type calcium channels recorded from bullfrog sympathetic neurons. AB - For many neurons, N-type calcium channels provide the primary pathway for calcium influx during an action potential. We investigated the gating properties of single N-type calcium channels using the cell-attached patch technique. With 100 mM Ba2+ in the pipet, mean N-channel open probability (Po, measured over 100 ms) increased with depolarization, but the range at a single voltage was large (e.g., Po at +40 mV ranged from 0.1 to 0.8). The open dwell time histograms were generally well fit by a single exponential with mean open time (tauo) increasing from 0.7 ms at +10 mV to 3.1 ms at +40 mV. Shut time histograms were well fit by two exponentials. The brief shut time component (taush1 = 0.3 ms) did not vary with the test potential, while the longer shut time component (taush2) decreased with voltage from 18.9 ms at +10 mV to 2.3 ms at +40 mV. Although N-channel Po during individual sweeps at +40 mV was often high ( approximately 0.8), mean Po was reduced by null sweeps, low Po gating, inactivation, and slow activation. The variability in mean Po across patches resulted from differences in the frequency these different gating processes were expressed by the channels. Runs analysis showed that null sweeps tended to be clustered in most patches, but that inactivating and slowly activating sweeps were generally distributed randomly. Low Po gating (Po = 0.2, tauo = 1 ms at +40 mV) could be sustained for approximately 1 min in some patches. The clustering of null sweeps and sweeps with low Po gating is consistent with the idea that they result from different modes of N-channel gating. While Po of the main N-channel gating state is high, the net Po is reduced to a maximum value of close to 0.5 by other gating processes. PMID- 9874693 TI - P2 receptor modulation of voltage-gated potassium currents in Brown adipocytes. AB - Using patch voltage-clamp techniques, we find there are two components to the voltage-gated potassium current (IKv) in rat brown adipocytes. The components differ in their gating and responses to purinergic stimulation, but not their pharmacology. IKv-A recovers from inactivation at physiological membrane potentials, while IKv-B inactivation recovers at more negative potentials. Both currents are >90% blocked by similar concentrations of quinine and tetraethylammonium, but not by beta-dendrotoxin, charybdotoxin, or apamin. The two current components are differentially modulated by extracellular ATP. ATP shifts the voltage dependence of IKv-A inactivation negative by 38 +/- 5 mV (n = 35, +/-SEM) and shifts activation by -14 +/- 2 mV in whole-cell experiments. ATP did not affect the steady state inactivation voltage dependence of IKv-B, but did apparently convert IKv-A into IKv-B. The pharmacology of the inactivation shift is consistent with mediation by a P2 purinergic receptor. Purinergic stimulation of perforated-patch clamped cells causes hyperpolarizing shifts in the window current of IKv-A by shifting inactivation -18 +/- 4 mV and activation -7 +/- 2 mV (n = 16). Since perforated-patch recordings will most closely resemble in vivo cell responses, this ATP-induced shift in the window current may facilitate IKv activation when the cell depolarizes. IKv activity is necessary for the proliferation and differentiation of brown adipocytes in culture (Pappone, P.A., and S.I. Ortiz-Miranda. 1993. Am. J. Physiol. 264:C1014-C1019) so purinergic modulation of IKv may be important in altering adipocyte growth and development. PMID- 9874694 TI - Voltage-insensitive gating after charge-neutralizing mutations in the S4 segment of Shaker channels. AB - Shaker channel mutants, in which the first (R362), second (R365), and fourth (R371) basic residues in the S4 segment have been neutralized, are found to pass potassium currents with voltage-insensitive kinetics when expressed in Xenopus oocytes. Single channel recordings clarify that these channels continue to open and close from -160 to +80 mV with a constant opening probability (Po). Although Po is low ( approximately 0.15) in these mutants, mean open time is voltage independent and similar to that of control Shaker channels. Additionally, these mutant channels retain characteristic Shaker channel selectivity, sensitivity to block by 4-aminopyridine, and are partially blocked by external Ca2+ ions at very negative potentials. Furthermore, mean open time is approximately doubled, in both mutant channels and control Shaker channels, when Rb+ is substituted for K+ as the permeant ion species. Such strong similarities between mutant channels and control Shaker channels suggests that the pore region has not been substantially altered by the S4 charge neutralizations. We conclude that single channel kinetics in these mutants may indicate how Shaker channels would behave in the absence of voltage sensor input. Thus, mean open times appear primarily determined by voltage-insensitive transitions close to the open state rather than by voltage sensor movement, even in control, voltage-sensitive Shaker channels. By contrast, the low and voltage-insensitive Po seen in these mutant channels suggests that important determinants of normal channel opening derive from electrostatic coupling between S4 charges and the pore domain. PMID- 9874695 TI - Pharmacokinetic considerations in obesity. PMID- 9874696 TI - Secondary structure and protein deamidation. AB - The deamidation reactions of asparagine residues in alpha-helical and beta-turn secondary structural environments of peptides and proteins are reviewed. Both kinds of secondary structure tend to stabilize asparagine residues against deamidation, although the effects are not large. The effect of beta-sheet structures on asparagine stability is unclear, although simple considerations suggest a stabilization in this environment also. PMID- 9874698 TI - MDCK (Madin-Darby canine kidney) cells: A tool for membrane permeability screening. AB - The goal of this work was to investigate the use of MDCK (Madin-Darby canine kidney) cells as a possible tool for assessing the membrane permeability properties of early drug discovery compounds. Apparent permeability (Papp) values of 55 compounds with known human absorption values were determined using MDCK cell monolayers. For comparison, Papp values of the same compounds were also determined using Caco-2 cells, a well-characterized in vitro model of intestinal drug absorption. Monolayers were grown on 0. 4-microm Transwell-COL membrane culture inserts. MDCK cells were seeded at high density and cultured for 3 days, and Caco-2 cells were cultured under standard conditions for 21 to 25 days. Compounds were tested using 100 microM donor solutions in transport medium (pH 7.4) containing 1% DMSO. The Papp values in MDCK cells correlated well with those in Caco-2 cells (r2 = 0.79). Spearman's rank correlation coefficient for MDCK Papp and human absorption was 0.58 compared with 0.54 for Caco-2 Papp and human absorption. These results indicate that MDCK cells may be a useful tool for rapid membrane permeability screening. PMID- 9874699 TI - Preparation and in vitro/in vivo evaluation of luteinizing hormone releasing hormone (LHRH)-loaded polyhedral and spherical/tubular niosomes. AB - Niosomes are vesicles formed by the self-assembly of nonionic surfactants in aqueous dispersions. They can entrap drugs and have been used experimentally as sustained drug delivery systems. Apart from conventional spherical niosomes, various types of vesicle ultrastructures can be formed by varying the composition of the vesicle membrane. Hexadecyl diglycerol ether (C16G2), cholesterol, and poly-24-oxyethylene cholesteryl ether (Solulan C24) in the ratio 91:0:9 gave polyhedral niosomes, whereas spherical and tubular niosomes are produced at a composition ratio of 49:49:2. The mean size of both polyhedral and spherical/tubular niosomes were within the range of 6 to 9 microm. Both types of vesicle were visualized by cryo-scanning electron microscopy. The properties of the two forms of niosomes were studied using luteinizing hormone releasing hormone (LHRH) as a model peptide. Analysis by high-performance liquid chromatography demonstrated high entrapment of LHRH acetate in polyhedral niosomes when prepared by remote loading methods using pH or (NH4)2SO4 gradients; in contrast, only low entrapment was achieved by passive loading methods (direct hydration at pH 7.4 or pH 3.0, dehydration-rehydration, and reversed-phase evaporation). In vitro studies demonstrated that both polyhedral and spherical/tubular niosomes were more stable in 5% rat skeletal muscle homogenate than in rat plasma. Also, polyhedral niosomes released more radiolabeled LHRH ([125I]LHRH) than spherical/tubular niosomes in both muscle homogenate and plasma. In clearance experiments in the rat, following intramuscular injection, both polyhedral and spherical/tubular niosomes gradually released [125I]LHRH into the blood, but some radioactivity remained at the injection site for 25 and 49 h, respectively. In contrast, [125I]LHRH in phosphate buffered saline was completely cleared from the injection site at 2 h. The release of drug is sustained by both niosome formulations, but spherical/tubular niosomes possess more stable membranes than polyhedral niosomes due to the presence of cholesterol. PMID- 9874700 TI - Solubility and mass and nuclear magnetic resonance spectroscopic studies on interaction of cyclosporin A with dimethyl-alpha- and -beta-cyclodextrins in aqueous solution. AB - The interaction of cyclosporin A (CsA) with dimethyl-alpha- and -beta cyclodextrins (DM-alpha-CyD and DM-beta-CyD) was investigated by the solubility method, electrospray ionization mass spectrometry (ESI-MS) and 1H-nuclear magnetic resonance spectroscopy (1H NMR). The extremely low solubility (1.9 x 10( 5) M at 25 degreesC) of CsA in water was significantly improved by the complexation with DM-CyDs: for example, the solubility increased 87-fold in the presence of 5.0 x 10(-2) M DM-beta-CyD. The phase solubility diagram of CsA/DM CyD systems showed an Ap type and the stability constants (1060 M-1 and 1050 M-1, respectively) of the 1:1 CsA/DM-alpha-CyD and CsA/DM-beta-CyD complexes were much higher than those of the 1:2 complexes (15 M-1 and 21 M-1, respectively). In ESI MS spectra of the CsA/DM-beta-CyD system, a new signal emerged at 1268 which corresponds to the 1:1 adduct of the di-ionized guest molecule with the host molecule. This signal intensity was significantly decreased by the addition of chlorpromazine (CPZ) which has a large stability constant (8800 M-1) of the DM beta-CyD complex, whereas the signal corresponding to the CPZ/DM-beta-CyD complex was little affected by the addition of CsA, indicating a competitive inclusion of CPZ and CsA within the host cavity. CsA gave many new peaks in the 1H NMR spectrum when the solvent was changed from chloroform to methanol/water, suggesting conformational diversity of CsA in polar solvents. Inspection of 1H chemical shift changes and the two-dimensional rotating frame nuclear Overhauser effect (ROESY) spectra of the CsA/DM-CyD system suggested that the side chains of amino acids in CsA molecule take part in the inclusion within DM-CyDs, although there is seemingly no preference of particular amino acid residues. All the data obtained here suggested that CsA forms inclusion complexes with DM-alpha- and beta-CyDs in an aqueous medium and side chains of CsA are mainly involved in the inclusion. PMID- 9874697 TI - Pharmacokinetic and metabolism studies using microdialysis sampling. PMID- 9874701 TI - Influence of gender on prednisolone effects on whole blood T-cell deactivation and trafficking in rats. AB - Prednisolone (5 mg/kg intravenous) was administered to adrenalectomized male and female Sprague-Dawley rats (250-350 g) to assess the effects of gender on disposition and pharmacoimmunodynamics. Plasma concentrations of prednisolone were determined by high-performance liquid chromatography. Incorporation of [3H]thymidine (3H-TDR) was used to determine whole blood T-cell (WBTC) trafficking and deactivation following stimulation with Concanavalin-A. Whole blood T-cell trafficking was determined indirectly by using the glucocorticoid receptor antagonist RU-40555 (250 ng/mL) added to ex vivo cultures of whole blood from animals dosed with prednisolone. Mean (+/- SD) prednisolone clearance values were 3.22 +/- 0.88 and 3.46 +/- 0.96 L/h/kg in males and females, respectively. After administration of prednisolone, relative T-cell counts decreased slowly with time to reach a nadir at 3-5 h and returned to baseline levels by 8 h. Fitting data using an indirect response model yielded mean prednisolone 50% inhibitory concentration for inhibition of WBTC trafficking (IC50T) that was lower in males compared with females (0.14 +/- 0.16 versus 1.03 +/- 0.06 ng/mL; p < 0.05). In the absence of RU-40555, an immediate and complete inhibition of 3H TDR incorporation into WBTC was observed (deactivation) and baseline levels were recovered slowly as prednisolone was cleared from blood. The mean 50% inhibitory concentration for inhibition of WBTC deactivation (IC50D) based on an inhibitory Imax model was similar in males and females (0.20 +/- 0.24 versus 0.18 +/- 0.12 ng/mL). Although male and female rats have similar exposure to prednisolone after 5-mg/kg doses, males are more sensitive to the inhibition of WBTC trafficking, whereas no gender effects on deactivation of WBTC exist. PMID- 9874702 TI - In vitro regioselective stability of beta-1-O- and 2-O-acyl glucuronides of naproxen and their covalent binding to human serum albumin. AB - beta-1-O- (NAG) and 2-O-glucuronides (2-isomer) of (S)-naproxen (NA) were prepared to determine which positional isomer(s) of the acyl glucuronide of NA is responsible for forming covalent adducts with human serum albumin (HSA). Their comparative stability and covalent binding adduct formation with HSA were investigated at pH 7.4 and at 37 degreesC. NA and its acyl glucuronides were simultaneously determined by HPLC. Three positional isomers were formed successively after incubation of NAG in the buffer only. However, when NAG was incubated with HSA (30 mg/mL), isomers other than the 2-isomer were formed in little or negligible quantities. In HSA solution, NAG (kd = 2.08 +/- 0.08 h-1) was four times less stable than 2-isomer (kd = 0.51 +/- 0.02 h-1). NAG was degraded by hydrolysis (khyd = 1.01 +/- 0.10 h-1) and isomerization (kiso = 1.07 +/- 0.07 h-1) to the same extent; however, hydrolysis was predominant for the 2 isomer (kd = 0.51 +/- 0.02 h-1). The incubation of both NAG and 2-isomer with HSA led to the formation of a covalent adduct; however, the adduct formation from the 2-isomer proceeded more slowly than that from NAG. The present results suggest that the covalent binding of NA to HSA via its acyl glucuronides proceeds through both transacylation (direct nucleophilic displacement) and glycation mechanisms; NAG rapidly forms an adduct that may be unstable, and the protein adduct from the 2-O-acyl glucuronide is as important for the covalent binding as those from the 1 O-acyl glucuronides. PMID- 9874703 TI - Conformational stability of lyophilized PEGylated proteins in a phase-separating system. AB - PEGylation of proteins is of great interest to the pharmaceutical industry as covalent attachment of poly(ethylene glycol) (PEG) molecules can increase protein sera half-lives and reduce antigenicity. Not surprisingly, PEGylation significantly alters the surface characteristics of a protein, and consequently, its conformational stability during freezing and drying. Freeze concentration induced phase separation between excipients has been previously shown to cause degradation of the secondary structure in lyophilized hemoglobin. In this report we show how PEGylation of two proteins, hemoglobin- and brain-derived neurotrophic factor (BDNF), influences partitioning and protein secondary structure as determined by FTIR spectroscopy in a system prone to freezing induced phase separation. PEGylation of hemoglobin reduces the loss of structure induced by lyophilization in a PEG/dextran system that phase separates during freezing, perhaps due to altered partitioning. The partition coefficient for native hemoglobin favors the dextran-rich phase (PEG/dextran partition coefficient = 0.3), while PEGylated hemoglobin favors the PEG phase (partition coefficient = 3.1). In addition, we demonstrate that PEGylation alters hemoglobin's stability during lyophilization in the absence of other excipients. In contrast, because native BDNF already partitions into the PEG-rich phase, PEGylation of BDNF has a less dramatic effect on both partition coefficients and conformational stability during lyophilization. This is the first report on the effects of PEGylation on protein structural stability during lyophilization and points out the need to consider modification of formulations in response to changing protein surface characteristics. PMID- 9874704 TI - A new model describing the swelling and drug release kinetics from hydroxypropyl methylcellulose tablets. AB - A novel mathematical model for the water transport into and drug release from hydroxypropyl methylcellulose (HPMC) tablets is presented. Fick's second law of diffusion is used to describe the mass transfer processes in the three-component system drug/polymer/water. Numerical solutions of the respective set of partial differential equations are provided, considering axial and radial diffusion within cylindrical tablets. It is shown that the diffusion coefficients strongly depend on the water concentration (parameters quantifying this dependence have been determined). Swelling of the device is considered using moving boundary conditions, whereas dissolution processes are neglected. Experiments proved the applicability of the theory. The practical benefit of the new model is to calculate the required shape and dimensions of HPMC tablets to achieve a desired release profile. PMID- 9874707 TI - Ionization constants and distribution coefficients of phenothiazines and calcium channel antagonists determined by a pH-metric method and correlation with calculated partition coefficients. AB - The pH-metric technique was used to determine the ionization constants and distribution coefficients of 10 phenothiazines and five ionizable calcium channel antagonists. Because the studied compounds were poorly water soluble and quite lipophilic with partition coefficients in the range of 3.5 to 5.5, organic cosolvents had to be added for the determination of the ionization constants to avoid precipitation of the free bases. The effect of the cosolvents dioxane and methanol on the extrapolation to pure water was compared. For both cosolvents a very good agreement with accessible published ionization constants was obtained, however the slope of the regression line was much smaller for dioxane, yielding more reliable estimates according to the standard deviation of the extrapolated values. Thus, dioxane might be preferable to methanol as a cosolvent for the determination of ionization constants of sparingly water soluble bases. Also the n-octanol/water partition coefficients were determined and compared with published data and values calculated with the ClogP, ACD, and HINT programs. Although the obtained values were approximate in conformity with the published data, the calculated partition coefficients differed from the experimental ones considerably for the majority of the investigated compounds. Furthermore, the ion pair partitioning and the distribution coefficients at physiological pH 7.4 were determined. The pH-dependent distribution profiles showed the strong influence of the ionization constants and of the distribution of the ion pairs on the overall distribution. This result strongly suggests that greater use should be made of measured distribution coefficients in quantitative structure-activity relationship studies. The potentiometric method is a convenient way to determine the distribution properties of drug molecules at pH values relevant for the biological system under investigation. PMID- 9874705 TI - Equilibrium and kinetics of rotamer interconversion in immunosuppressant prodigiosin derivatives in solution. AB - The equilibrium and relative rate of rotamer interconversion around the bond joining the 2,2'-bipyrrolyl and pyrromethene moieties in a synthetic analogue of immunosuppressant prodigiosin are investigated as a function of pHapp in a water/acetonitrile mixture (1/1 by volume). Two chromatographically separable isomeric forms are obtained in acid solutions (pHapp < 4), whereas rapid interconversion occurs above neutrality. Furthermore, pH modulates the conformational preference of the molecule according to nitrogen protonation on the three pyrrole rings system (pKa = 7.2). At high pHapp (neutral form), the same conformer that is observed in pure acetonitrile prevails, whereas the other one is preferred by the protonated form. The nuclear magnetic resonance data indicate that the structures of the two conformers mainly differ in the value of the torsion angle around the aforementioned C-C bond. Kinetic and equilibrium data are quantitatively interpreted with a cyclic mechanism including two protonation (pKa1 = 8.23 +/- 0.03, pKa2 = 5. 4 +/- 0.2) and two conformational rearrangement steps. A molecular interpretation of the observed behavior includes, for the preferred conformer at low pH, formation of a new hydrogen bond between the exocyclic oxygen and the neighboring pyrrole NH upon protonation of the three pyrrole rings system. PMID- 9874706 TI - Acute and long-term stability studies of deoxy hemoglobin and characterization of ascorbate-induced modifications. AB - The reaction of ascorbate with recombinant hemoglobin (rHb1.1) in the presence of differing partial pressures of oxygen was studied. In the presence of 15 000 ppm (1.5%) residual oxygen, ascorbate/oxygen-mediated reactions resulted in an increased rate of autoxidation, modification of the beta-globin, increased oxygen affinity and decreased maximum Hill coefficient. One of the observed modifications to the beta-globin was a 72 Da addition to its N-terminus. Detailed characterization indicates the modification was an imidazolidinone type structure. Thorough deoxygenation of the hemoglobin solution to <150 ppm of oxygen prior to addition of ascorbate was required to prevent these modifications. Addition of ascorbate to the deoxy hemoglobin (deoxyHb) at pH 8 induced aggregation, eventually leading to precipitation. No such precipitation was observed at pH 7. Long-term storage of the hemoglobin was carried out by addition of ascorbate to deoxyHb at pH 7. The level of methemoglobin remained at <2% for up to 1 year at 4 degreesC, with no detectable precipitation of the protein. Modifications similar to those observed by the acute studies were observed over the 1-year period and correlated with disappearance of the added ascorbate. PMID- 9874708 TI - Intravenous pretreatment with empty pH gradient liposomes alters the pharmacokinetics and toxicity of doxorubicin through in vivo active drug encapsulation. AB - Liposomes have been used widely to improve the therapeutic activity of pharmaceutical agents. The traditional approach for such applications has been to formulate the pharmaceutical agent in liposomes prior to administration in vivo. In this report we demonstrate that liposomes exhibiting a transmembrane pH gradient injected intravenously (iv) can actively encapsulate doxorubicin in the circulation after iv administration of free drug. Small (110 nm) liposomes composed of phosphatidylcholine (PC)/cholesterol (Chol, 55:45 mol:mol) exhibiting a pH gradient (inside acidic) were administered iv 1 h prior to free doxorubicin, and plasma drug levels as well as toxicity and efficacy were evaluated. Predosing with egg PC/Chol pH gradient liposomes increased the plasma concentration of doxorubicin as much as 200-fold compared to free drug alone as well as to predosing with dipalmitoyl PC/Chol pH gradient liposomes or EPC/Chol liposomes without a pH gradient. The ability of the liposomes to alter the pharmacokinetics of doxorubicin was dependent on the presence of a transmembrane pH gradient and correlated with the extent of doxorubicin uptake into the liposomes at 37 degreesC in pH 7.5 buffer, indicating that doxorubicin was being actively accumulated in the circulating liposomes. This in vivo drug loading was achieved over a range of doxorubicin doses (5 mg/kg-40 mg/kg) and was dependent on the dose of EPC/Chol liposomes administered prior to free doxorubicin injection. The altered pharmacokinetic properties of doxorubicin associated with in vivo doxorubicin encapsulation were accompanied by a decrease in drug toxicity and maintained antitumor potency. These results suggest that pretreatment with empty liposomes exhibiting a pH gradient may provide a versatile and straightforward method for enhancing the pharmacological properties of many drugs that can accumulate into such vesicle systems at physiological temperatures. PMID- 9874709 TI - Demonstration of the terms enantiotropy and monotropy in polymorphism research exemplified by flurbiprofen. AB - The thermodynamic terms enantiotropy and monotropy are demonstrated by means of solid-state analytical results of polymorphous flurbiprofen (FBP). Vibrational spectra, differential scanning calorimetry (DSC), and thermomicroscopy investigations as well as X-ray powder patterns for three modifications of FBP are described. The melting points are mod. I 113-114 degreesC (enthalpy of fusion 27.9 +/- 0.2 kJ mol-1) for modification I (mod. I), 92 degreesC for mod. II, and 87 degreesC for mod. III. The true densities of mod. I (1.279 +/- 0.001 g cm-3) and mod. II (1.231 +/- 0.002 g cm-3) were measured at 25 degreesC. Modification I (commercial product) is the thermodynamically stable crystal form from absolute zero to its melting point. Modification II was crystallized on a gram scale from a warm saturated solution of FBP in n-heptane and rapid cooling of the solution to -18 degreesC. Modification I is monotropically related to mod. II and mod. III, due to application of the density rule and the entropy-of-fusion rule. The thermodynamic relationships between the three modifications are demonstrated by a semischematic energy/temperature diagram. Theoretical vapor pressure/temperature diagrams and energy/temperature diagrams are compared and briefly discussed. PMID- 9874710 TI - Estimation of aqueous solubility for some guanine derivatives using partition coefficient and melting temperature. AB - Aqueous solubilities for some guanine derivatives were estimated by semiempirical equations developed by Yalkowsky and Valvani1 using the data for partition coefficient and melting temperature. It was shown that in the case of guanine derivatives examined in this study, the solubility values could not be estimated adequately by these equations. PMID- 9874711 TI - Importance of tautomers in the chemical behavior of tetracyclinesdagger. AB - We advance the concept that tautomerism is crucial for the understanding of the chemical behavior of tetracycline. Indeed, considering four deprotonations, there are 64 different possible tautomers to be considered for tetracycline. Our results indicate that tetracycline is a very adaptive molecule, capable of easily modifying itself through tautomerism in response to various chemical environments. Indeed, its situation in solution can be more accurately pictured as an equilibrium among a diversity of tautomeric species-an equilibrium that can be easily displaced depending on the various possible chemical perturbations, such as varying the pH or the dielectric constant of the solvent. Moreover, we also show that tetracycline could undergo four deprotonations and predict for it a fourth pKa of 13 and refer to our experimental determination of this parameter, which yielded the value of 12. We conclude that tautomerism is essential to the comprehension of the chemical behavior of tetracycline as determined by the semiempirical method AM1 as well as by the self-consistent reaction field method, which estimates the effects of the solvent on the tautomers. All tautomers in their different conformations have been fully optimized for each of the possible degrees of protonation of this molecule. Thus, the relative stabilities of the different tautomeric species have been computed. PMID- 9874712 TI - Pharmacokinetics of thalidomide in an elderly prostate cancer population. AB - Thalidomide, a glutamic acid derivative, has recently been shown to inhibit in vitro angiogenesis, the process of formation of new blood vessels. This Phase II study examined the pharmacokinetics of thalidomide in patients with clinically progressive hormone-refractory prostate cancer. Patients (aged 55 to 80 years) were randomized to two different arms, low dose versus high dose. Patients in the low-dose group were given 200 mg of thalidomide and patients in the high-dose group received 200 mg of thalidomide, with subsequent dose escalations to 1200 mg. Serial serum or blood samples were obtained for pharmacokinetic assessment after administration of a single oral dose or multiple daily dosing of thalidomide and were assayed by reversed-phase HPLC. Pharmacokinetic parameters for both the single and multiple dosing were calculated with ADAPT II. A one compartment model best fit the data. After single dosing, the oral clearance and apparent volume of distribution for the low-dose regimen (n = 13) were 7.41 +/- 2.05 L/h and 66.93 +/- 34.27 L, respectively, whereas for the high-dose regimen (n = 11), these values were 7.21 +/- 2.89 L/h and 165.81 +/- 84.18 L, respectively. The elimination half-lives for the low and high dose were 6.52 +/- 3.81 and 18.25 +/- 14.08 h, respectively. After the multiple dosing of thalidomide, the oral clearance and apparent volume of distribution for the low dose group (n = 10) were 6.35 +/- 1.64 L/h and 64.63 +/- 23.20 L, respectively, whereas for the high-dose group (n = 11), these values were 7.73 +/- 2.27 L/h and 167.85 +/- 82.08 L, respectively. The elimination half-lives for the low and high dose were 7.08 +/- 1.87 and 16.19 +/- 9.57 h, respectively. For both the single and multiple dosing of thalidomide, the apparent volume of distribution and half life were significantly higher for the high-dose group than those for the low dose group. The higher apparent volume of distribution may be attributable to several factors, such as change in absorption, protein binding, etc. A dose proportional increase in thalidomide steady-state concentrations was seen after multiple daily dosing of thalidomide. PMID- 9874713 TI - PLGA microspheres containing plasmid DNA: preservation of supercoiled DNA via cryopreparation and carbohydrate stabilization. AB - Biodegradable microspheres containing plasmid DNA have potential uses as mediators of transfection in cells, particularly phagocytic cells such as macrophages. However, the hydrophilic nature and the structural instability of supercoiled DNA preclude its facile encapsulation in polymer matrixes such as poly(d, l-lactic-co-glycolic acid) (PLGA) by traditional methods. We initially studied the microencapsulation of plasmid DNA using the established water-in-oil in-water double-emulsion solvent-evaporation method and found that (1) the encapsulation efficiency was low (about 20%), (2) the microencapsulation procedure nicked (degraded) the supercoiled DNA, and (3) lyophilization of the microsphere also nicked the DNA. We have therefore designed a new microsphere preparation method (called cryopreparation) to specifically address these concerns. Using the cryopreparation method, the aqueous phase of the primary emulsion containing the plasmid DNA is frozen and then subjected to homogenization. Because there is no shear stress inside a solid, we hypothesized that freezing the aqueous phase of the primary emulsion would help to preserve the supercoiled plasmid DNA during formation of the secondary emulsion. We also hypothesized that the formation of crystals from buffers within the primary emulsion was a causative factor for nicking during freezing or lyophilization, and that disruption of the crystal formation by the addition of saccharides into the primary emulsion would improve the supercoiled-DNA content of the spheres. Our results support the two hypotheses. Not only was the supercoiled-DNA content increased from 39% to over 85%, but the encapsulation efficiency was also elevated from 23% to over 85%. PMID- 9874714 TI - Effective administration route for the deleted form of hepatocyte growth factor To exert its pharmacological effects. AB - The pharmacokinetics and the pharmacological effects of the deleted form of hepatocyte growth factor (dHGF) after intravenous (iv), subcutaneous (sc), or intramuscular (im) administration (0.25 and 2. 5 mg/kg) were studied in rats. After single iv administration (2.5 mg/kg), dHGF in serum rapidly decreased (alpha- and beta-phase half-life: 3.2 and 26.5 min, respectively). Two to four hours after single sc or im administration (2.5 mg/kg), the serum level of dHGF reached a maximum and then gradually declined (half-life: 2.7 h). The serum levels were not changed by repetitive iv administration, but were dramatically decreased by repetitive sc or im administration. Liver weight and serum levels of total protein, albumin, and HDL-cholesterol were significantly increased by iv administration of dHGF (twice daily for 4 days at 0.25 mg/kg). Sc or im administration of dHGF did not increase these parameters at the same dose, but did significantly at 2.5 mg/kg. These observations suggest that iv administration is the most effective in exerting the pharmacological effects of dHGF among three administration routes. dHGF after iv administration was distributed mainly and rapidly into liver (53.6% of the injected dHGF within 5 min) and was sustained at a higher level in the liver than in plasma. In infusion (0.5 mg/kg/3 h), dHGF level in plasma and liver reached a steady-state 15 and 60 min after starting the infusion, respectively. The steady-state level of dHGF was 7- to 9-fold higher in liver than in plasma, and the higher level in liver was sustained beyond the steady-state. PMID- 9874715 TI - Microparticles of novel branched copolymers of lactic acid and amino acids: preparation and characterization. AB - The preparation and characterization of microparticles produced from a new class of functionalized, biodegradable, comblike graft copolymers is presented. The copolymers are polyester-polyamino acid hybrids, composed of a poly(L-lactic acid co-L-lysine) (PLAL) backbone, and poly(L-lysine), poly(D,L-alanine) or poly(L aspartic acid) side chains extending from the lysine residues of PLAL. The microparticles have been characterized with regard to their surface properties, morphology, and size. Thus, electron spectroscopy for chemical analysis data and results of Zeta potential measurements suggest that the polyamino acid side chains tend to concentrate at the surface of the particles. Also, analyses by environmental scanning electron microscopy and confocal scanning laser microscopy indicate that particles carrying poly(lysine) chains have an unusual porous structure, most probably due to the combined effects of the amphiphilic, polyelectrolyte, and chemical nature of the composing copolymer, as well as of the particular preparation technique employed. The capabilities of the microparticles to serve as carriers in controlled drug release and delivery devices were demonstrated by encapsulation and release of rhodamine B, a low molecular weight drug model. PMID- 9874716 TI - Carrier-mediated absorption of salicylic acid from hamster cheek pouch mucosa. AB - Previously, we found that monocarboxylic acids undergo carrier-mediated transport in primary cultures of oral mucosal epithelial cells.1 In this study, we investigated whether carrier-mediated absorption of a monocarboxylic acid from the oral mucosa occurs in vivo. Salicylic acid was administered to hamster cheek pouch. At predetermined intervals, the concentration of salicylic acid in the fluid remaining in the cheek pouch lumen and the blood salicylic acid concentration were determined. The absorption of salicylic acid was saturable at high salicylic acid concentrations. Sodium azide, a metabolic inhibitor, and carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP), a protonophore, significantly inhibited the absorption of salicylic acid but not the absorption of salicylamide from the oral mucosa. Various monocarboxylic acids inhibited the absorption of salicylic acid, whereas dicarboxylic acids had no such effect. Transfer of [14C]salicylic acid from the cheek pouch mucosa to the systemic circulation was observed, and the blood [14C]salicylic acid concentration in the case of coadministration with propionic acid was significantly lower than that in the case of no propionic acid coadministration. These results show that monocarboxylic acids undergo carrier-mediated absorption from the hamster cheek pouch mucosa. PMID- 9874717 TI - Pharmacokinetics and organ distribution of cationized colchicine-specific IgG and Fab fragments in rat. AB - Pharmacokinetics of cationized goat colchicine-specific polyclonal immunoglobulin G (IgG) and antigen binding fragment (Fab) (cIgG and cFab, respectively) were studied in male adult Sprague-Dawley rats and compared with those of the native proteins (nIgG and nFab). All proteins were radioiodinated by the Iodogen method, and kinetics were investigated following trichloroacetic acid (TCA) precipitation or immunoprecipitation. Deiodination and catabolism were more pronounced with the cationized than the native proteins, especially for cFab. Both cIgG and cFab in plasma decreased more rapidly than nIgG and nFab. The elimination half-lives were 52.9 and 81.8 h for cIgG and nIgG, respectively. In addition, there was a 74-fold increase in the volume of distribution and a 114-fold increase in the systemic clearance of cIgG compared with nIgG. For cFab, the volume of distribution and systemic clearance were increased 6.4- and 3.5-fold, respectively. Organ uptake of cIgG and cFab was markedly increased compared with that of nIgG and nFab, especially in kidney, liver, spleen, and lung. Renal clearance of cIgG and cFab was also increased 30- and 10-fold compared with that of nIgG and nFab, respectively. The present data suggest that cationization of colchicine-specific IgG and Fab fragments increased the organ distribution and greatly altered their pharmacokinetics. Nevertheless, the smaller molecular size of Fab versus IgG did not enhance the distribution and clearance of cFab. These data pave the way for evaluating the biological efficacy of these more tissue-organ-interactive antibodies. PMID- 9874718 TI - Artificial neural networks applied to the in vitro-in vivo correlation of an extended-release formulation: initial trials and experience. AB - Artificial neural networks applied to in vitro-in vivo correlations (ANN-IVIVC) have the potential to be a reliable predictive tool that overcomes some of the difficulties associated with classical regression methods, principally, that of providing an a priori specification of the regression equation structure. A number of unique ANN configurations are presented, that have been evaluated for their ability to determine an IVIVC from different formulations of the same product. Configuration variables included a combination of architectural structures, learning algorithms, and input-output association structures. The initial training set consisted of two formulations and included the dissolution from each of the six cells in the dissolution bath as inputs, with associated outputs consisting of 1512 pharmacokinetic time points from nine patients enrolled in a crossover study. A third formulation IVIVC data set was used for predictive validation. Using these data, a total of 29 ANN configurations were evaluated. The ANN structures included the traditional feed forward, recurrent, jump connections, and general regression neural networks, with input-output association types consisting of the direct mapping of the dissolution profiles to the pharmacokinetic observations, mapping the individual dissolution points to the individual observations, and using a "memorative" input-output association. The ANNs were evaluated on the basis of their predictive performance, which was excellent for some of these ANN models. This work provides a basic foundation for ANN-IVIVC modeling and is the basis for continued modeling with other desirable inputs, such as formulation variables and subject demographics. PMID- 9874729 TI - THE PLANT VACUOLE. AB - Plant cells are unique in containing large acidic vacuoles which occupy most of the cell volume. The vacuolar H+-ATPase (V-ATPase) is the enzyme responsible for acidifying the central vacuole, although it is also present on Golgi and coated vesicles. Many secondary transport processes are driven by the proton-motive force generated by the V-ATPase, including reactions required for osmoregulation, homeostasis, storage, plant defense and many other functions. However, a second proton pump, the V-PPase, serves as a potential back-up system and may, in addition, pump potassium. The plant V-ATPase is structurally similar to other eukaryotic V-ATPases and its subunits appear to be encoded by small multigene families. These multigene families may play important roles in the regulation of gene expression and in the sorting of V-ATPase isoforms to different organelles. PMID- 9874745 TI - INSECT ION HOMEOSTASIS. AB - The constant composition of body fluids in insects is maintained by the cooperative interaction of gastrointestinal and urinary tissues. Water follows ionic movements, which are driven by the basolateral Na+/K+-ATPase and/or the apical 'K+(or Na+) pump'. The latter now is thought to be the functional expression of a parallel arrangement of a proton-motive V-ATPase and a K+(or Na+)/nH+ antiport. This review focuses on the pathways for the movement of monovalent inorganic ions through epithelia involved in ion homeostasis. A graphical summary compares the principal findings with respect to cation secretion in lepidopteran caterpillar midgut goblet cells (K+) and in brush border cells of Malpighian tubules (K+, Na+). PMID- 9874730 TI - VACUOLAR-TYPE H+-TRANSLOCATING ATPases IN PLANT ENDOMEMBRANES: SUBUNIT ORGANIZATION AND MULTIGENE FAMILIES. AB - Acidification of endomembrane compartments by the vacuolar-type H+-translocating ATPase (V-ATPase) is vital to the growth and development of plants. The V-ATPase purified from oat roots is a large complex of 650x10(3 )Mr that contains 10 different subunits of 70, 60, 44, 42, 36, 32, 29, 16, 13 and 12x10(3 )Mr. This set of ten polypeptides is sufficient to couple ATP hydrolysis to proton pumping after reconstitution of the ATPase into liposomes. Unlike some animal V-ATPases, the purified and reconstituted V-ATPase from oat is directly stimulated by Cl-. The peripheral complex of the ATPase includes the nucleotide-binding subunits of 70 and 60x10(3 )Mr and polypeptides of 44, 42, 36 and 29x10(3 )Mr. Six copies of the 16x10(3 )Mr proteolipid together with three other polypeptides are thought to make up the integral sector that forms the H+-conducting pathway. Release of the peripheral complex from the native membrane completely inactivates the pump; however, the peripheral subunits can be reassembled with the membrane sector to form a functional H+ pump. Comparison of V-ATPases from several plants indicates considerable variations in subunit composition. Hence, several forms of the V ATPase may exist among, and probably within, plant species. At least four distinct cDNAs encode the 16x10(3 )Mr proteolipid subunit in oat. Multiple genes could encode different subtypes of the H+ pump that are regulated by the developmental stage and physiological function specific to the cell or tissue type. PMID- 9874747 TI - THE INSECT V-ATPase, A PLASMA MEMBRANE PROTON PUMP ENERGIZING SECONDARY ACTIVE TRANSPORT: IMMUNOLOGICAL EVIDENCE FOR THE OCCURRENCE OF A V-ATPase IN INSECT ION TRANSPORTING EPITHELIA. AB - Active electrogenic K+ transport in insects serves as the energy source for secretion or absorption in gastrointestinal epithelia or for the receptor current in sensory epithelia. In the larval midgut of the tobacco hornworm Manduca sexta, a vacuolar-type proton pump (V-ATPase) and a K+/nH+ antiport represent the functional elements of the potassium pump. Several immunological findings support the hypothesis that active K+ transport in other insect epithelia may also be energized by a V-ATPase. In immunoblots, crude homogenates of sensilla-rich antennae and Malpighian tubules of M. sexta cross reacted with an immune serum directed to the purified plasma membrane V-ATPase from the midgut; the M. sexta midgut V-ATPase cross reacted with polyclonal antibodies to endomembrane V ATPases from xenic origin. In immunocytochemical investigations of larvae of M. sexta and adults of Antheraea pernyi, monoclonal antibodies to defined subunits of the purified midgut V-ATPase or polyclonal antibodies to xenic endomembrane V ATPase labelled the sites of active K+ transport: the goblet cell apical membrane in the midgut, the brush border of Malpighian tubules and the apical projections of the auxiliary cells in antennal sensilla. The functional mechanism of a primary H+-pumping V-ATPase and a secondary H+-dependent K+ transport postulated for K+-transporting insect epithelia may be further applicable to active Na+ or Cl- transport and would provide a unifying concept for all ouabain-insensitive electrogenic ion transport in insects. The findings from the midgut investigations, however, are the first instance in which a V-ATPase provides an alternative to the Na+/K+-ATPase in energizing secondary active transport in animal plasma membranes. PMID- 9874748 TI - pH GRADIENTS IN LEPIDOPTERAN MIDGUT. AB - Lepidopteran larvae demonstrate several remarkable specialisations of the alimentary canal: the most active epithelial transport known; a unique cell type, called a goblet cell; and the highest pH values known to be generated by a biological system. The electrogenic K+ pump in midgut is now known to be energised by a H+-pumping V-ATPase, and net alkali metal transport is achieved by linking it to a nH+/alkali metal exchanger, which recycles H+ into the cytoplasm. Generation of high luminal pH is modelled as a passive (Nernstian) distribution of protons in the electrical field generated by the V-type ATPase. Electrode impalements show that the potential difference across the goblet cavity membrane is extremely high. Measurements of pH gradients generated in vitro confirm that the midgut itself generates such a gradient, that this process relies on metabolic energy, and that the differential ability of midgut subregions to perform acid-base transport maps to their differing morphologies and to the pH profiles observed along the gut in vivo. During larval/larval moults, K+ transport is suppressed. The transepithelial potential difference (PD) across the gut collapses and recovers in phase with the loss and recovery of the gut pH gradient, and with tissue V-ATPase activity, confirming that these processes are intimately linked. Acridine Orange partitions into acidic compartments and might be expected to be concentrated in goblet cavities, as these are the compartments toward which the V-ATPase pumps protons. However, under normal conditions, Acridine Orange is excluded from the cavities. Red metachromasia of the cavities (implying low pH) is only observed when the ion transport status of the tissue is compromised. It thus seems likely that, under physiological conditions, K+/H+ exchange is tight enough to produce a neutral or alkaline, rather than acidic, cavity. Molecular analysis of the 16 000 Mr subunit from Manduca midgut reveals it to be closely similar to other known 16 000 Mr sequences, particularly that from Drosophila brain. It is thus likely to be a true H+ channel, rather than one modified for K+ transport. The cavity can be modelled in two ways: (i) to isolate the site of proton equilibration electrically from the main gut lumen, and thus allow larger pH gradients to develop, or (ii) to buffer the V-ATPase from the alkaline pH in the gut lumen, which would otherwise destroy the gradient driving the exchange of H+ for alkali metal cations. The first model would predict a high cavity pH, whereas the second would predict a near neutral pH and would imply a non-cavity route for transport of base equivalents. Work with both pH-sensitive dyes and pH-sensitive electrodes so far tends to support the second model. PMID- 9874752 TI - INSECT MALPIGHIAN TUBULES: V-ATPase ACTION IN ION AND FLUID TRANSPORT. AB - Insect Malpighian tubules secrete fluid into the lumen as part of their function as excretory organs. The underlying ion transport is, when stimulated, faster than in any other known tissue. It is driven by the activity of an H+ transporting V-ATPase situated on the luminal cell membranes. This ATPase, together with cation/H+ antiporter(s), constitutes a common cation pump which can transport sodium ions, potassium ions or both. Treatments that selectively slow cation transport across the epithelium cause the secreted fluid to become alkaline, whereas those that selectively reduce the rate of anion passage lead to secretion of acid fluid. PMID- 9874753 TI - A PLANT BIOCHEMIST'S VIEW OF H+-ATPases AND ATP SYNTHASES. AB - My twenty-five year fascination with membrane ATPases grew out of my experiences in the laboratories of Andre Jagendorf and Efraim Racker. Andre introduced me to photosynthetic phosphorylation and Ef, to whose memory this article is dedicated, convinced me that ATPases had much to do with ATP synthesis. Astounding progress has been made in the H+-ATPase field in just two decades. By the early 1970s, it was generally recognized that oxidative and photosynthetic ATP synthesis were catalyzed by membrane enzymes that could act as H+-ATPases and that the common intermediate between electron transport and phosphorylation is the electrochemical proton gradient. At that time, it had been shown that a cation stimulated ATPase activity was associated with plasma membrane preparations from plant roots. The endomembrane or vacuolar ATPases were unknown. The application of improved biochemical methods for membrane isolation and purification, as well as membrane protein reconstitutions, led rapidly to the conclusion that there are three major classes of membrane H+-ATPases, P, V and F. P-ATPases, which will not be considered further in this article, are phosphorylated during their catalytic cycle and have a much simpler polypeptide composition than V- or F-ATPases. The plasma membrane H+-ATPase of plant, yeasts and fungal cells is one example of this class of enzymes (see Pedersen and Carafoli, 1987, for a comparison of plasma membrane ATPases). Biochemical and gene sequencing analysis have revealed that V- and F-ATPases resemble each other structurally, but are distinct in function and origin. The 'V' stands for vacuolar and the 'F' for F1Fo. F1 was the first factor isolated from bovine heart mitochondria shown to be required for oxidative phosphorylation. Fo was so named because it is a factor that conferred oligomycin sensitivity to soluble F1. Other F-ATPases are often named to indicate their sources. For example, chloroplast F1 is denoted CF1 (see Racker, 1965, for early work on F1). Recent successes in reconstitution of vacuolar ATPase have led to a V1Vo nomenclature for this enzyme as well. The term 'ATP synthase' is now in general use to describe F-ATPases. This term emphasizes the facts that although F ATPases function to synthesize ATP, they do not catalyze, normally, ATP hydrolysis linked to proton flux. In contrast, V-ATPases are very unlikely to operate as ATP synthases. Thus, F-ATPases are proton gradient consumers, whereas V-ATPases generate proton gradients at the expense of hydrolysis. In this brief review, I will compare the structures of F- and V-ATPases. Also, I give some insight into the mechanisms that help prevent wasteful ATP hydrolysis by the chloroplast ATP synthase (CF1Fo). PMID- 9874756 TI - THE CHLOROPLAST H+-ATPase: PARTIAL REACTIONS OF THE PROTON. AB - This article reviews proton intake, charge transfer and proton release by F ATPases, based in part on flash spectrophotometric studies on the chloroplast ATPase in thylakoid membranes, CF1Fo. The synthesis-coupled translocation of charges by CF1Fo (maximum rate <1500 s-1) and the dissipative flow through its exposed channel portion, CFo (rate >10 000 s-1), are extremely proton-specific (selectivity H+:K+>10(7):1). The proton-specific filter is located in CFo. Proton flow through exposed CFo can be throttled by adding subunit (&dgr;) or subunit &bgr; of CF1. These subunits thus may provide energy-transducing contacts between CF1 and CFo. Recently, we characterized two conditions where, in contrast to the above situation, proton intake by CF1Fo was decoupled from proton transfer across the main dielectric barrier: (a) CF1Fo structurally distorted by low ionic strength transiently trapped protons in a highly cooperative manner, but remained proton tight. This result has been interpreted in terms of Mitchell's proton well. (b) In the absence of nucleotides there is a proton slip. Addition of nucleotides (100 nmol l-1 ADP) abolished proton conduction but not proton intake by CF1Fo. These experiments functionally tag proton binding groups on CF1Fo that are located before the main dielectric barrier. PMID- 9874757 TI - HALOBACTERIAL A-ATP SYNTHASE IN RELATION TO V-ATPase. AB - The head piece separated from the A-ATP synthase of Halobacterium halobium hydrolyses ATP. This A1-ATPase is inhibited by nitrate but not by other chaotropic anions. The nitrate inhibition is noncompetitive with respect to ATP, reversible, and partially protected by chloride. In contrast, ATP synthase in situ (A1Ao-ATPase) is not inhibited by nitrate but apparently is inhibited by stronger chaotropic reagents, such as thiocyanate and trichloroacetate, which make the vesicle membrane permeable to protons. The mode of action of nitrate and chaotropic anions seems to differentiate A-ATPases from V-ATPases. Other strains of Halobacterium, Haloferax, Haloarcula, Halococcus and Natronobacterium, contain at least two polypeptides immunochemically similar to the two major subunits, (&agr;) (86x10(3 )Mr on SDS-PAGE) and &bgr; (64x10(3 )Mr), of the A-ATPase of Halobacterium halobium. When solubilized, membrane vesicles of these halobacteria hydrolyse ATP. Their ATPases are commonly sensitive to nitrate. They require high concentrations of the supporting salt but depend differently on chloride or sulfate/sulfite. The A-ATPases of Halobacteriaceae appear to diverge with respect to salt preference. PMID- 9874759 TI - Transcriptional regulation: contending with complexity. PMID- 9874761 TI - Coevolutionary arms races: is victory possible? PMID- 9874760 TI - Why is Hu where? Shuttling of early-response-gene messenger RNA subsets. PMID- 9874762 TI - A microfabricated device for sizing and sorting DNA molecules. AB - We have demonstrated a microfabricated single-molecule DNA sizing device. This device does not depend on mobility to measure molecule size, is 100 times faster than pulsed-field gel electrophoresis, and has a resolution that improves with increasing DNA length. It also requires a million times less sample than pulsed field gel electrophoresis and has comparable resolution for large molecules. Here we describe the fabrication and use of the single-molecule DNA sizing device for sizing and sorting DNA restriction digests and ladders spanning 2-200 kbp. PMID- 9874763 TI - Enhancement of cellulose production by expression of sucrose synthase in Acetobacter xylinum. AB - Higher plants efficiently conserve energy ATP in cellulose biosynthesis by expression of sucrose synthase, in which the high free energy between glucose and fructose in sucrose can be conserved and used for the synthesis of UDP-glucose. A mixture of sucrose synthase and bacterial cellulose synthase proceeded to form UDP-glucose from sucrose plus UDP and to synthesize 1,4-beta-glucan from the sugar nucleotide. The mutant sucrose synthase, which mimics phosphorylated sucrose synthase, enhanced the reaction efficiency (Vmax/Km) on 1,4-beta-glucan synthesis, in which the incorporation of glucose from sucrose was increased at low concentrations of UDP. Because UDP formed after glucosyl transfer can be directly recycled with sucrose synthase, UDP-glucose formed appears to show high turnover with cellulose synthase in the coupled reaction. The expression of sucrose synthase in Acetobacter xylinum not only changed sucrose metabolism but also enhanced cellulose production, in which UDP-glucose was efficiently formed from sucrose. Although the level of UDP-glucose in the transformant with mutant sucrose synthase cDNA was only 1.6-fold higher than that in plasmid-free cells, the level of UDP was markedly decreased in the transformant. The results show that sucrose synthase serves to channel carbon directly from sucrose to cellulose and recycles UDP, which prevents UDP build-up in cellulose biosynthesis. PMID- 9874764 TI - The oncogenic transforming potential of the passage of single alpha particles through mammalian cell nuclei. AB - Domestic, low-level exposure to radon gas is considered a major environmental lung-cancer hazard involving DNA damage to bronchial cells by alpha particles from radon progeny. At domestic exposure levels, the relevant bronchial cells are very rarely traversed by more than one alpha particle, whereas at higher radon levels-at which epidemiological studies in uranium miners allow lung-cancer risks to be quantified with reasonable precision-these bronchial cells are frequently exposed to multiple alpha-particle traversals. Measuring the oncogenic transforming effects of exactly one alpha particle without the confounding effects of multiple traversals has hitherto been unfeasible, resulting in uncertainty in extrapolations of risk from high to domestic radon levels. A technique to assess the effects of single alpha particles uses a charged-particle microbeam, which irradiates individual cells or cell nuclei with predefined exact numbers of particles. Although previously too slow to assess the relevant small oncogenic risks, recent improvements in throughput now permit microbeam irradiation of large cell numbers, allowing the first oncogenic risk measurements for the traversal of exactly one alpha particle through a cell nucleus. Given positive controls to ensure that the dosimetry and biological controls were comparable, the measured oncogenicity from exactly one alpha particle was significantly lower than for a Poisson-distributed mean of one alpha particle, implying that cells traversed by multiple alpha particles contribute most of the risk. If this result applies generally, extrapolation from high-level radon risks (involving cellular traversal by multiple alpha particles) may overestimate low level (involving only single alpha particles) radon risks. PMID- 9874765 TI - CIR, a corepressor linking the DNA binding factor CBF1 to the histone deacetylase complex. AB - CBF1 is a member of the CSL family of DNA binding factors, which mediate either transcriptional repression or transcriptional activation. CSL proteins play a central role in Notch signaling and in Epstein-Barr virus-induced immortalization. Notch is a transmembrane protein involved in cell-fate decisions, and the cytoplasmic domain of Notch (NotchIC) targets CBF1. The Epstein-Barr virus-immortalizing protein EBNA2 activates both cellular and viral gene expression by targeting CBF1 and mimicking NotchIC. We have examined the mechanism of CBF1-mediated repression and show that CBF1 binds to a unique corepressor, CBF1 interacting corepressor (CIR). A CIR homolog is encoded by Caenorhabditis elegans, indicating that CIR is evolutionarily conserved. Two CBF1 mutants that were unable to bind CIR did not function as repressors, suggesting that targeting of CIR to CBF1 is an important component of repression. When expressed as a Gal4 fusion protein, CIR repressed reporter gene expression. CIR binds to histone deacetylase and to SAP30 and serves as a linker between CBF1 and the histone deacetylase complex. PMID- 9874766 TI - A change in actin conformation associated with filament instability after Pi release. AB - The ability of actin to both polymerize into filaments and to depolymerize permits the rapid rearrangements of actin structures that are essential for actin's function in most cellular processes. Filament polarity and dynamic properties are conferred by the hydrolysis of ATP on actin filaments. Release of inorganic phosphate (Pi) from filaments after ATP hydrolysis promotes depolymerization. We identify a yeast actin mutation, Val-159 to Asn, which uncouples Pi release from the conformational change that results in filament destabilization. Three-dimensional reconstructions of electron micrographs reveal a conformational difference between ADP-Pi filaments and ADP filaments and show that ADP V159N filaments resemble ADP-Pi wild-type filaments. Crystal structures of mammalian beta-actin in which the nucleotide binding cleft is in the "open" and "closed" states can be used to model actin filaments in the ADP and ADP-Pi conformations, respectively. We propose that these two conformations of G-actin may be related to two functional states of F-actin. PMID- 9874767 TI - Proton exit from the heme-copper oxidase of Escherichia coli. AB - Pathways of proton entry have been identified in the proton-translocating heme copper oxidases, but the proton exit pathway is unknown. Here we report experiments with cytochrome bo3 in Escherichia coli cells that may identify the beginning of the exit pathway. Systematic mutations of arginines 438 and 439 (R481 and R482 in the E. coli enzyme), numbering as in cytochrome aa3 from bovine heart mitochondria, which interact with the ring D propionates of the two heme groups, reveal that the D propionate of the oxygen-binding heme is involved in proton pumping; its anionic form must be stabilized in order for proton translocation to occur. This may locate the beginning of the pathway by which pumped protons exit from the enzyme structure. PMID- 9874768 TI - Mutagenesis of conserved lysine residues in bacteriophage T5 5'-3' exonuclease suggests separate mechanisms of endo-and exonucleolytic cleavage. AB - Efficient cellular DNA replication requires the activity of a 5'-3' exonuclease. These enzymes are able to hydrolyze DNA.DNA and RNA.DNA substrates exonucleolytically, and they are structure-specific endonucleases. The 5'-3' exonucleases are conserved in organisms as diverse as bacteriophage and mammals. Crystal structures of three representative enzymes identify two divalent-metal binding sites typically separated by 8-10 A. Site-directed mutagenesis was used to investigate the roles of three lysine residues (K83, K196, and K215) situated near two metal-binding sites in bacteriophage T5 5'-3' exonuclease. Neither K196 nor K215 was essential for either the exo- or the endonuclease activity, but mutation of these residues increased the dissociation constant for the substrate from 5 nM to 200 nM (K196A) and 50 nM (K215A). Biochemical analysis demonstrated that K83 is absolutely required for exonucleolytic activity on single-stranded DNA but is not required for endonucleolytic cleavage of flap structures. Structural analysis of this mutant by x-ray crystallography showed no significant perturbations around the metal-binding sites in the active site. The wild-type protein has different pH optima for endonuclease and exonuclease activities. Taken together, these results suggest that different mechanisms for endo- and exonucleolytic hydrolysis are used by this multifunctional enzyme. PMID- 9874769 TI - Catalytic properties of selenophosphate synthetases: comparison of the selenocysteine-containing enzyme from Haemophilus influenzae with the corresponding cysteine-containing enzyme from Escherichia coli. AB - The selD gene from Haemophilus influenzae has been overexpressed in Escherichia coli. The expressed protein was purified to homogeneity in a four-step procedure and then carboxymethylated by reaction with chloroacetate. N-terminal sequencing by Edman degradation identified residue 16 as carboxymethyl selenocysteine, which corresponded to the essential cysteine residue in the glycine-rich sequence of the E. coli selenophosphate synthetase. It would be expected that an ionized selenol of a selenocysteine in place of a catalytically essential cysteine residue would result in an enzyme with increased catalytic activity. To test this hypothesis we kinetically characterized the selenocysteine containing selenophosphate synthetase from H. influenzae and compared its catalytic activity to that of the cysteine containing selenophosphate synthetase from E. coli. Our characterization revealed the Km values for the two substrates, selenide and ATP, were similar for both enzymes. However, the selenocysteine-containing enzyme did not exhibit the expected higher catalytic activity. Based on these results we suggest a role of selenocysteine in H. influenzae that is not catalytic. PMID- 9874770 TI - Mechanisms by which IkappaB proteins control NF-kappaB activity. AB - The biological activity of the transcription factor NF-kappaB is differentially controlled by three IkappaB proteins, Ikappa Balpha, Ikappa Bbeta, and Ikappa Bepsilon. We have examined the molecular basis for the differential inhibitory strengths of IkappaB proteins by constructing hybrid IkappaBs and found that the first ankyrin repeat of Ikappa Balpha is responsible for its strong inhibitory effect. Swapping a putative beta-turn within the first ankyrin repeat between the strong Ikappa Balpha and the weak IkappaBbeta inhibitors switches their in vivo inhibitory activity on NF-kappaB. The qualitatively distinct contacts made by this beta-turn in Ikappa Balpha and Ikappa Bbeta with NF-kappaB determine the efficiency by which IkappaBs sequester NF-kappaB to the cytoplasm, thus explaining their distinct effects on gene activity. PMID- 9874771 TI - Homotypic interaction and multimerization of nucleocapsid protein of tomato spotted wilt tospovirus: identification and characterization of two interacting domains. AB - The nucleocapsid protein (N) of tomato spotted wilt tospovirus (TSWV) plays a central role in the viral life cycle. With the aid of the yeast two-hybrid system and surface plasmon resonance analysis, homotypic interaction and multimerization of the N protein was detected. Analysis of deletion mutants identified two binding regions in the protein, located at the N terminus (amino acids 1-39) and the C terminus (amino acids 233-248), respectively, implying a "head-to-tail" interaction of the N terminus with the C terminus to form a multimeric chain. Further characterization of the binding domains was performed by site-directed mutagenesis. Two phenylalanines (F242 and F246) highly conserved in the N proteins within the Tospovirus genus were shown to play a crucial role in the interaction. PMID- 9874772 TI - An arrayable flow-through microcentrifuge for high-throughput instrumentation. AB - A compact, flow-through centrifugation system has been developed specifically for high-throughput centrifugation of large numbers of samples. The instrument is based on multiple high-speed rotors that also serve as sample holders. The small size of the rotors allows them to be arrayed in a standard 96-well microtiter plate spacing, making this device ideal for highly parallel automated instrumentation. Though initially designed for cell separation in DNA sequencing protocols, the flow-through centrifuge can be used to replace conventional centrifugation in most processes involving small samples. Techniques for recovery of both the supernatant and the pellet have been developed, as well as techniques for sample mixing, and cleaning of the reusable rotors. This paper discusses the design and performance of the flow-through centrifuge applied to cell separation and resuspension and to DNA purification and concentration. PMID- 9874773 TI - Mediator protein mutations that selectively abolish activated transcription. AB - Deletion of any one of three subunits of the yeast Mediator of transcriptional regulation, Med2, Pgd1 (Hrs1), and Sin4, abolished activation by Gal4-VP16 in vitro. By contrast, other Mediator functions, stimulation of basal transcription and of TFIIH kinase activity, were unaffected. A different but overlapping Mediator subunit dependence was found for activation by Gcn4. The genetic requirements for activation in vivo were closely coincident with those in vitro. A whole genome expression profile of a Deltamed2 strain showed diminished transcription of a subset of inducible genes but only minor effects on "basal" transcription. These findings make an important connection between transcriptional activation in vitro and in vivo, and identify Mediator as a "global" transcriptional coactivator. PMID- 9874774 TI - Mechanism of recruitment of DnaB helicase to the replication origin of the plasmid pSC101. AB - Although many bacterial chromosomes require only one replication initiator protein, e.g., DnaA, most plasmid replicons depend on dual initiators: host encoded DnaA and plasmid-encoded Rep initiator protein for replication initiation. Using the plasmid pSC101 as a model system, this work investigates the biological rationale for the requirement for dual initiators and shows that the plasmid-encoded RepA specifically interacts with the replicative helicase DnaB. Mutations in DnaB or RepA that disrupt RepA-DnaB interaction cause failure to load DnaB to the plasmid ori in vitro and to replicate the plasmid in vivo. Although, interaction of DnaA with DnaB could not substitute for RepA-DnaB interaction for helicase loading, DnaA along with integration host factor, DnaC, and RepA was essential for helicase loading. Therefore, DnaA is indirectly needed for helicase loading. Instead of a common surface of interaction with initiator proteins, interestingly, DnaB helicase appears to have at least a limited number of nonoverlapping surfaces, each of which interacts specifically with a different initiator protein. PMID- 9874775 TI - Cytosolic adenylyl cyclase defines a unique signaling molecule in mammals. AB - Mammals have nine differentially regulated isoforms of G protein-responsive transmembrane-spanning adenylyl cyclases. We now describe the existence of a distinct class of mammalian adenylyl cyclase that is soluble and insensitive to G protein or Forskolin regulation. Northern analysis indicates the gene encoding soluble adenylyl cyclase (sAC) is preferentially expressed in testis. As purified from rat testis cytosol, the active form of sAC appears to be a fragment derived from the full-length protein, suggesting a proteolytic mechanism for sAC activation. The two presumptive catalytic domains of sAC are closely related to cyanobacterial adenylyl cyclases, providing an evolutionary link between bacterial and mammalian signaling molecules. PMID- 9874776 TI - Characterization of functionally active subribosomal particles from Thermus aquaticus. AB - Peptidyl transferase activity of Thermus aquaticus ribosomes is resistant to the removal of a significant number of ribosomal proteins by protease digestion, SDS, and phenol extraction. To define the upper limit for the number of macromolecular components required for peptidyl transferase, particles obtained by extraction of T. aquaticus large ribosomal subunits were isolated and their RNA and protein composition was characterized. Active subribosomal particles contained both 23S and 5S rRNA associated with notable amounts of eight ribosomal proteins. N terminal sequencing of the proteins identified them as L2, L3, L13, L15, L17, L18, L21, and L22. Ribosomal protein L4, which previously was thought to be essential for the reconstitution of particles active in peptide bond formation, was not found. These findings, together with the results of previous reconstitution experiments, reduce the number of possible essential macromolecular components of the peptidyl transferase center to 23S rRNA and ribosomal proteins L2 and L3. Complete removal of ribosomal proteins from T. aquaticus rRNA resulted in loss of tertiary folding of the particles and inactivation of peptidyl transferase. The accessibility of proteins in active subribosomal particles to proteinase hydrolysis was increased significantly after RNase treatment. These results and the observation that 50S ribosomal subunits exhibited much higher resistance to SDS extraction than 30S subunits are compatible with a proposed structural organization of the 50S subunit involving an RNA "cage" surrounding a core of a subset of ribosomal proteins. PMID- 9874777 TI - Gene delivery: a single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus. AB - Translocation of exogenous DNA through the nuclear membrane is a major concern of gene delivery technologies. To take advantage of the cellular import machinery, we have synthesized a capped 3.3-kbp CMVLuciferase-NLS gene containing a single nuclear localization signal peptide (PKKKRKVEDPYC). Transfection of cells with the tagged gene remained effective down to nanogram amounts of DNA. Transfection enhancement (10- to 1,000-fold) as a result of the signal peptide was observed irrespective of the cationic vector or the cell type used. A lysine to threonine mutation of the third NLS amino acid completely abolished these remarkable features, suggesting importin-mediated translocation. Our hypothesis is that the 3-nm-wide DNA present in the cytoplasm is initially docked to and translocated through a nuclear pore by the nuclear import machinery. As DNA enters the nucleus, it is quickly condensed into a chromatin-like structure, which provides a mechanism for threading the remaining worm-like molecule through the pore. A single NLS signal is thus sufficient, whereas many signals on a gene would actually inhibit entry, the same DNA molecule being threaded through adjacent pores. PMID- 9874778 TI - Pneumocysterol [(24Z)-ethylidenelanost-8-en-3beta-ol], a rare sterol detected in the opportunistic pathogen Pneumocystis carinii hominis: structural identity and chemical synthesis. AB - Pneumocystis carinii pneumonia (PcP) remains among the most prevalent opportunistic infections among AIDS patients. Currently, drugs used clinically for deep mycosis act by binding ergosterol or disrupting its biosynthesis. Although classified as a fungus, P. carinii lacks ergosterol. Instead, the pathogen synthesizes a number of distinct Delta7, 24-alkylsterols, despite the abundance of cholesterol, which it can scavenge from the lung alveolus. Thus, the pathogen-specific sterols appear vital for organism survival and proliferation. In the present study, high concentrations of a C32 sterol were found in human derived P. carinii hominis. The definitive structural identities of two C-24 alkylated lanosterol compounds, previously not reported for rat-derived P. carinii carinii, were determined by using GLC, MS, and NMR spectroscopy together with the chemical syntheses of authentic standards. The C31 and C32 sterols were identified as euphorbol (24-methylenelanost-8-en-3beta-ol) and pneumocysterol [(24Z)-ethylidenelanost-8-en-3beta-ol], respectively. The identification of these and other 24-alkylsterols in P. carinii hominis suggests that (i) sterol C-24 methyltransferase activities are extraordinarily high in this organism, (ii) 24 alkylsterols are important components of the pathogen's membranes, because the addition of these side groups onto the sterol side chain requires substantial ATP equivalents, and (iii) the inefficacy of azole drugs against P. carinii can be explained by the ability of this organism to form 24-alkysterols before demethylation of the lanosterol nucleus. Because mammals cannot form 24 alkylsterols, their biosyntheses in P. carinii are attractive targets for the development of chemotherapeutic strategies against this opportunistic infection. PMID- 9874779 TI - Disordered water within a hydrophobic protein cavity visualized by x-ray crystallography. AB - Water in the hydrophobic cavity of human interleukin 1beta, which was detected by NMR spectroscopy but was invisible by high resolution x-ray crystallography, has been mapped quantitatively by measurement and phasing of all of the low resolution x-ray diffraction data from a single crystal. Phases for the low resolution data were refined by iterative density modification of an initial flat solvent model outside the envelope of the atomic model. The refinement was restrained by the condition that the map of the difference between the electron density distribution in the full unit cell and that of the atomic model be flat within the envelope of the well ordered protein structure. Care was taken to avoid overfitting the diffraction data by maintaining phases for the high resolution data from the atomic model and by a resolution-dependent damping of the structure factor differences between data and model. The cavity region in the protein could accommodate up to four water molecules. The refined solvent difference map indicates that there are about two water molecules in the cavity region. This map is compatible with an atomic model of the water distribution refined by using XPLOR. About 70% of the time, there appears to be a water dimer in the central hydrophobic cavity, which is connected to the outside by two constricted channels occupied by single water molecules approximately 40% of the time on one side and approximately 10% on the other. PMID- 9874780 TI - Maturation of the tyrosine kinase c-src as a kinase and as a substrate depends on the molecular chaperone Hsp90. AB - Although Hsp90 displays general chaperone activity in vitro, few substrates of the chaperone have been identified in vivo, and the characteristics that render these substrates dependent on Hsp90 remain elusive. To investigate this issue, we exploited a paradoxical observation: several unrelated oncogenic viral tyrosine kinases, including v-src, attain their native conformation after association with Hsp90, yet their nearly identical cellular homologs interact only weakly with the chaperone. It has been controversial whether Hsp90 is vital for normal maturation of the cellular kinases or is simply binding a misfolded subfraction of the proteins. By modulating Hsp90 levels in Saccharomyces cerevisiae, we determined that Hsp90 is indeed necessary for the maturation of c-src (the normal homolog of v-src). c-src maturation is, however, less sensitive to Hsp90 perturbations than is v-src maturation. Dependence of the two proteins on Hsp90 does not correspond to their relative efficiency in reaching their final destination (the plasma membrane); we observed that in yeast, unlike in vertebrate cells, neither c-src nor v-src concentrate in the membrane. Expression of different v/c-src chimeras in cells carrying wild-type or temperature-sensitive Hsp90 alleles revealed that the difference between the proteins instead arises from multiple, naturally occurring mutations in the C-terminal region of v-src. PMID- 9874781 TI - Centrosomal control of microtubule dynamics. AB - In many animal cells, minus ends of microtubules (MTs) are thought to be capped by the centrosome whereas plus ends are free and display dynamic instability. We tested the role of the centrosome by examining MT behavior in cytoplasts from which the centrosome was removed. Cells were injected with Cy3-tubulin to fluorescently label MTs and were enucleated by using a centrifugation procedure. Enucleation resulted in a mixture of cytoplasts containing or lacking the centrosome. Fibroblast (CHO-K1) and epithelial (BSC-1) cells were investigated. In fibroblast cytoplasts containing the centrosome, MTs showed dynamic instability indistinguishable from that in intact cells. In contrast, in cytoplasts lacking the centrosome, MTs treadmilled-shortened at the minus end at about 12 micrometers/min while growing at the plus end at the same rate. The change in behavior of the plus end from dynamic instability to persistent growth correlated with an elevated level of free tubulin subunits (78% in centrosome free cytoplasts vs. 44% in intact cells) generated by minus-end depolymerization. In contrast to fibroblast cells, in centrosome-free cytoplasts prepared from epithelial cells, MTs displayed dynamic instability at plus ends and relative stability at minus ends presumably because of specific minus-end stability factors distributed throughout the cytoplasm. We suggest that, in fibroblast cells, a minus-end depolymerization mechanism functions to eliminate errors in MT organization and that dynamic instability of MT plus ends is a result of capping of minus ends by the centrosome. PMID- 9874783 TI - Transient and stable gene expression in mammalian cells transduced with a recombinant baculovirus vector. AB - Recombinant baculoviruses can serve as gene-transfer vehicles for transient expression of recombinant proteins in a wide range of mammalian cell types. Furthermore, by inclusion of a dominant selectable marker in the viral vector, cell lines can be derived that stably express recombinant genes. A virus was constructed containing two expression cassettes controlled by constitutive mammalian promoters: the cytomegalovirus immediate early promoter/enhancer directing expression of green fluorescent protein and the simian virus 40 (SV40) early promoter controlling neomycin phosphotransferase II. Using this virus, efficient gene delivery and expression was observed and measured in numerous cell types of human, primate, and rodent origin. In addition to commonly used transformed cell lines such as HeLa, CHO, Cos-7, and 293, this list includes primary human keratinocytes and bone marrow fibroblasts. In all cases, addition of butyrate or trichostatin A (a selective histone deacetylase inhibitor) to transduced cells markedly enhanced the levels of reporter protein expression observed. When transduced cells are put under selection with the antibiotic G418, cell lines can be obtained at high frequency that stably maintain the expression cassettes of the vector DNA and exhibit stable, high-level expression of the reporter gene. Stably transduced derivatives have been selected from a substantial number of different cell types, suggesting that stable lines can be derived from any cell type that exhibits transient expression. PMID- 9874782 TI - Calcium influx factor is synthesized by yeast and mammalian cells depleted of organellar calcium stores. AB - Depletion of endoplasmic reticulum Ca2+ stores leads to the entry of extracellular Ca2+ into the cytoplasm, a process termed capacitative or store operated Ca2+ entry. Partially purified extracts were prepared from the human Jurkat T lymphocyte cell line and yeast in which Ca2+ stores were depleted by chemical and genetic means, respectively. After microinjection into Xenopus laevis oocytes, the extracts elicited a wave of increased cytoplasmic free Ca2+ ([Ca2+]i) that spread from the point of injection across the oocyte. Extracts from cells with replete organellar Ca2+ stores were inactive. The increases depended on extracellular Ca2+, were unaffected by the inositol 1,4,5 trisphosphate (IP3) inhibitor heparin or an anti-IP3 receptor antibody and were unchanged when the endoplasmic reticulum was segregated to the hemisphere opposite the injection site by centrifugation. Confocal microscopy revealed that [Ca2+]i increases were most pronounced at the periphery of the oocyte. The patterns of [Ca2+]i increases were replicated by computer simulations based on a diffusible messenger of about 700 Da that directly activates Ca2+ influx. In addition, ICRAC, a Ca2+ release-activated Ca2+ current monitored in Jurkat cells by whole-cell patch clamp recordings, was more rapidly activated when active extracts were included in the patch pipette than by the inclusion of a Ca2+ chelator or IP3. These data support the existence in yeast and mammalian cells depleted of Ca2+ stores of a functionally conserved diffusible calcium influx factor that directly activates Ca2+ influx. PMID- 9874784 TI - Alendronate mechanism of action: geranylgeraniol, an intermediate in the mevalonate pathway, prevents inhibition of osteoclast formation, bone resorption, and kinase activation in vitro. AB - Nitrogen-containing bisphosphonates were shown to cause macrophage apoptosis by inhibiting enzymes in the biosynthetic pathway leading from mevalonate to cholesterol. This study suggests that, in osteoclasts, geranylgeranyl diphosphate, the substrate for prenylation of most GTP binding proteins, is likely to be the crucial intermediate affected by these bisphosphonates. We report that murine osteoclast formation in culture is inhibited by both lovastatin, an inhibitor of hydroxymethylglutaryl CoA reductase, and alendronate. Lovastatin effects are blocked fully by mevalonate and less effectively by geranylgeraniol whereas alendronate effects are blocked partially by mevalonate and more effectively by geranylgeraniol. Alendronate inhibition of bone resorption in mouse calvaria also is blocked by mevalonate whereas clodronate inhibition is not. Furthermore, rabbit osteoclast formation and activity also are inhibited by lovastatin and alendronate. The lovastatin effects are prevented by mevalonate or geranylgeraniol, and alendronate effects are prevented by geranylgeraniol. Farnesol and squalene are without effect. Signaling studies show that lovastatin and alendronate activate in purified osteoclasts a 34-kDa kinase. Lovastatin-mediated activation is blocked by mevalonate and geranylgeraniol whereas alendronate activation is blocked by geranylgeraniol. Together, these findings support the hypothesis that alendronate, acting directly on osteoclasts, inhibits a rate-limiting step in the cholesterol biosynthesis pathway, essential for osteoclast function. This inhibition is prevented by exogenous geranylgeraniol, probably required for prenylation of GTP binding proteins that control cytoskeletal reorganization, vesicular fusion, and apoptosis, processes involved in osteoclast activation and survival. PMID- 9874785 TI - Nuclear endpoint of Wnt signaling: neoplastic transformation induced by transactivating lymphoid-enhancing factor 1. AB - The interaction between beta-catenin and LEF-1/TCF transcription factors plays a pivotal role in the Wnt-1 signaling pathway. The level of beta-catenin is regulated by partner proteins, including glycogen synthase kinase-3beta (GSK 3beta) and the adenomatous polyposis coli (APC) tumor suppressor protein. Genetic defects in APC are responsible for a heritable predisposition to colon cancer. APC protein and GSK-3beta bind beta-catenin, retain it in the cytoplasm, and facilitate the proteolytic degradation of beta-catenin. Abrogation of this negative regulation allows beta-catenin to translocate to the nucleus and to form a transcriptional activator complex with the DNA-binding protein lymphoid enhancing factor 1 (LEF-1). This complex is thought to be involved in tumorigenesis. Here we show that covalent linkage of LEF-1 to beta-catenin and to transcriptional activation domains derived from the estrogen receptor or the herpes simplex virus protein VP16 generates transcriptional regulators that induce oncogenic transformation of chicken embryo fibroblasts. The chimeras between LEF-1 and beta-catenin or VP16 are constitutively active, whereas fusions of LEF-1 to the estrogen receptor are regulatable by estrogen. These experiments document the oncogenicity of transactivating LEF-1 and show that the transactivation domain normally provided by beta-catenin can be replaced by heterologous activation domains. These results suggest that the transactivating function of the LEF-1/beta-catenin complex is critical for tumorigenesis and that this complex transforms cells by activating specific LEF-1 target genes. PMID- 9874786 TI - Proapoptotic activity of Caenorhabditis elegans CED-4 protein in Drosophila: implicated mechanisms for caspase activation. AB - CED-4 protein plays an important role in the induction of programmed cell death in Caenorhabditis elegans through the activation of caspases. However, the precise mechanisms by which it activates caspases remain unknown. To investigate the conservation of CED-4 function in evolution, transgenic Drosophila lines that express CED-4 in the compound eye were generated. Ectopic expression of CED-4 in the eyes induced massive apoptotic cell death through caspase activation. An ATP binding site (P-loop) mutation in CED-4 (K165R) causes a loss of function in its ability to activate Drosophila caspase, and an ATPase inhibitor blocks the CED-4 dependent caspase activity in Drosophila S2 cells. Immunoprecipitation analysis showed that both CED-4 and CED-4 (K165R) bind directly to Drosophila caspase drICE, and the overexpression of CED-4 (K165R) inhibits CED-4-, ecdysone-, or cycloheximide-dependent caspase activation in S2 cells. Furthermore, CED-4 (K165R) partially prevented cell death induced by CED-4 in Drosophila compound eyes. Thus, CED-4 function is evolutionarily conserved in Drosophila, and the molecular mechanisms by which CED-4 activates caspases might require ATP binding and direct interaction with the caspases. PMID- 9874787 TI - A bioluminescence resonance energy transfer (BRET) system: application to interacting circadian clock proteins. AB - We describe a method for assaying protein interactions that offers some attractive advantages over previous assays. This method, called bioluminescence resonance energy transfer (BRET), uses a bioluminescent luciferase that is genetically fused to one candidate protein, and a green fluorescent protein mutant fused to another protein of interest. Interactions between the two fusion proteins can bring the luciferase and green fluorescent protein close enough for resonance energy transfer to occur, thus changing the color of the bioluminescent emission. By using proteins encoded by circadian (daily) clock genes from cyanobacteria, we use the BRET technique to demonstrate that the clock protein KaiB interacts to form homodimers. BRET should be particularly useful for testing protein interactions within native cells, especially with integral membrane proteins or proteins targeted to specific organelles. PMID- 9874788 TI - Imaging of multicellular large-scale rhythmic calcium waves during zebrafish gastrulation. AB - Oscillations of cytosolic free calcium levels have been shown to influence gene regulation and cell differentiation in a variety of model systems. Intercellular calcium waves thus present a plausible mechanism for coordinating cellular processes during embryogenesis. Herein we report use of aequorin and a photon imaging microscope to directly observe a rhythmic series of intercellular calcium waves that circumnavigate zebrafish embryos over a 10-h period during gastrulation and axial segmentation. These waves first appeared at about 65% epiboly and continued to arise every 5-10 min up to at least the 16-somite stage. The waves originated from loci of high calcium activity bordering the blastoderm margin. Several initiating loci were active early in the wave series, whereas later a dorsal marginal midline locus predominated. On completion of epiboly, the dorsal locus was incorporated into the developing tail bud and continued to generate calcium waves. The locations and timing at which calcium dynamics are most active appear to correspond closely to embryonic cellular and syncytial sites of known morphogenetic importance. The observations suggest that a panembryonic calcium signaling system operating in a clock-like fashion might play a role during vertebrate axial patterning. PMID- 9874789 TI - Placental failure in mice lacking the homeobox gene Dlx3. AB - Dlx3 is a homeodomain transcription factor and a member of the vertebrate Distal less family. Targeted deletion of the mouse Dlx3 gene results in embryonic death between day 9.5 and day 10 because of placental defects that alter the development of the labyrinthine layer. In situ hybridization reveals that the Dlx3 gene is initially expressed in ectoplacental cone cells and chorionic plate, and later in the labyrinthine trophoblast of the chorioallantoic placenta, where major defects are observed in the Dlx3 -/- embryos. The expression of structural genes, such as 4311 and PL-1, which were used as markers to follow the fate of different derivatives of the placenta, was not affected in the Dlx3-null embryos. However, by day 10.5 of development, expression of the paired-like homeodomain gene Esx1 was strongly down-regulated in affected placenta tissue, suggesting that Dlx3 is required for the maintenance of Esx1 expression, normal placental morphogenesis, and embryonic survival. PMID- 9874790 TI - Self-incompatibility alleles from Physalis: implications for historical inference from balanced genetic polymorphisms. AB - Balanced genetic polymorphism has been proposed as a source from which to infer population history complementary to that of neutral genetic polymorphism, because genetic polymorphism maintained by balancing selection permits inferences about population size over much longer spans of time. However, empirical data for both S genes and major histocompatibility complex genes do not fit expectations of coalescent theory. Species-specific gene genealogies have longer terminal branches than expected, indicating an apparent slowdown in the origination of new alleles. Here, we present evidence that divergent S alleles were selectively maintained in Physalis cinerascens during a reduction in population size, generating longer terminal branches in the S gene genealogy relative to the congener Physalis crassifolia. Retention of divergent alleles during reduction in the number of alleles violates assumptions of the coalescent model used to estimate effective population size. Recent theoretical and empirical results are consistent with the proposition that nonrandom sorting is a general property of balanced genetic polymorphisms, suggesting that studies of balanced polymorphism that infer the absence of population bottlenecks may overestimate effective population size. PMID- 9874791 TI - Emergence of a dual-catalytic RNA with metal-specific cleavage and ligase activities: the spandrels of RNA evolution. AB - In vitro selection, or directed molecular evolution, allows the isolation and amplification of rare sequences that satisfy a functional-selection criterion. This technique can be used to isolate novel ribozymes (RNA enzymes) from large pools of random sequences. We used in vitro evolution to select a ribozyme that catalyzes a novel template-directed RNA ligation that requires surprisingly few nucleotides for catalytic activity. With the exception of two nucleotides, most of the ribozyme contributes to a template, suggesting that it is a general prebiotic ligase. More surprisingly, the catalytic core built from randomized sequences actually contains a 7-nt manganese-dependent self-cleavage motif originally discovered in the Tetrahymena group I intron. Further experiments revealed that we have selected a dual-catalytic RNA from random sequences: the RNA promotes both cleavage at one site and ligation at another site, suggesting two conformations surrounding at least one divalent metal ion-binding site. Together, these results imply that similar catalytic RNA motifs can arise under fairly simple conditions and that multiple catalytic structures, including bifunctional ligases, can evolve from very small preexisting parts. By breaking apart and joining different RNA strands, such ribozymes could have led to the production of longer and more complex RNA polymers in prebiotic evolution. PMID- 9874792 TI - Polyglutamine-mediated dysfunction and apoptotic death of a Caenorhabditis elegans sensory neuron. AB - The effect of expressing human huntingtin fragments containing polyglutamine (polyQ) tracts of varying lengths was assessed in Caenorhabditis elegans ASH sensory neurons in young and old animals. Expression of a huntingtin fragment containing a polyQ tract of 150 residues (Htn-Q150) led to progressive ASH neurodegeneration but did not cause cell death. Progressive cell death and enhanced neurodegeneration were observed in ASH neurons that coexpressed Htn-Q150 and a subthreshold dose of a toxic OSM-10::green fluorescent protein (OSM 10::GFP) fusion protein. Htn-Q150 huntingtin protein fragments formed protein aggregates in ASH neurons, and the number of ASH neurons containing aggregates increased as animals aged. ASH neuronal cell death required ced-3 caspase function, indicating that the observed cell death is apoptotic. Of interest, ced 3 played a critical role in Htn-Q150-mediated neurodegeneration but not in OSM10::GFP-mediated ASH neurodegeneration. ced-3 function was important but not essential for the formation of protein aggregates. Finally, behavioral assays indicated that ASH neurons, coexpressing Htn-Q150 and OSM10::GFP, were functionally impaired at 3 days before the detection of neurodegeneration, cell death, and protein aggregates. PMID- 9874794 TI - Formation of coding joints in V(D)J recombination-inducible severe combined immune deficient pre-B cell lines. AB - Characterization of the severe combined immune deficient (scid) defect in the recombination process has provided many insights into the underlying mechanisms of variable (diversity) joining recombination. By using recombination-inducible scid pre-B cell lines transformed with the temperature-sensitive Abelson-murine leukemia virus, we show that large quantities of recombination intermediates can be generated, and their resolution can be followed during further cell culture. In this study, we demonstrate that the ability of these scid pre-B cell lines to resolve coding ends depends on the cell culture temperature. At the nonpermissive temperature of 39 degreesC, scid pre-B cell lines fail to form coding joints and contain mostly unresolved hairpin-coding ends. Once the cell culture is returned to the permissive temperature of 33 degreesC, these same cells make a significant amount of coding joints concomitant with the disappearance of hairpin-coding ends. Thus, the scid cells are capable of resolving coding ends under certain culture conditions. However, the majority of the recovered coding joints contains extensive deletions, indicating that the temperature-dependent resolution of coding ends is still scid-like. Our results suggest that the inability of scid cells to promptly nick hairpin-coding ends may lead to aberrant joining in these cells. PMID- 9874793 TI - A two-step, two-signal model for the primary activation of precursor helper T cells. AB - I present here a new model for the primary activation of precursor helper T cells. Observations demonstrate that the immune system learns not to respond to extrathymic, organ-specific self-antigens because of their early appearance in development. The immune system thus discriminates between peripheral self antigens and foreign antigens and, when mature, usually makes an immune response against only the latter. Contemporary models for the activation and inactivation of T helper (Th) function do not account for such discrimination. The model proposed here is consistent with contemporary findings and incorporates a mechanism of peripheral self-nonself discrimination. PMID- 9874796 TI - The BCR-ABL oncoprotein potentially interacts with the xeroderma pigmentosum group B protein. AB - The previously uncharacterized CDC24 homology domain of BCR, which is missing in the P185 BCR-ABL oncogene of Philadelphia chromosome (Ph1)-positive acute lymphocytic leukemia but is retained in P210 BCR-ABL of chronic myelogeneous leukemia, was found to bind to the xeroderma pigmentosum group B protein (XPB). The binding appeared to be required for XPB to be tyrosine-phosphorylated by BCR ABL. The interaction not only reduced both the ATPase and the helicase activities of XPB purified in the baculovirus system but also impaired XPB-mediated cross complementation of the repair deficiency in rodent UV-sensitive mutants of group 3. The persistent dysfunction of XPB may in part underlie genomic instability in blastic crisis. PMID- 9874795 TI - A small number of residues in the class II molecule I-Au confer the ability to bind the myelin basic protein peptide Ac1-11. AB - The N-terminal peptide Ac1-11 of myelin basic protein induces experimental autoimmune encephalomyelitis in H-2(u) and (H-2(u) x H-2(s)) mice but does not in H-2(s) mice. Ac1-11 binds weakly to the class II major histocompatibility complex (MHC) molecule I-Au but not at all to I-As. We have studied the interaction of Ac1-11 and I-Au as a model system for therapeutic intervention in the autoimmune response seen in experimental autoimmune encephalomyelitis. Two polymorphic residues that differ between I-Au and I-As, Y26beta and T28beta, and one conserved residue, E74beta, confer specific binding of Ac1-11 to I-Au. A fourth residue, R70beta in I-Au, affects both peptide binding and T cell recognition. These results are consistent with a model that places arginine at position five of Ac1-11 in pockets 4 and 7 of the MHC groove, which is formed in part by residues 26, 28, 70, and 74 of Abetau and places lysine at position four of Ac1 11, previously shown to be a major MHC contact, in hydrophobic pocket 6. The data indicate that the primary region of I-Au that confers specific binding of Ac1-11 lies in the center of the peptide binding groove rather than in the region that contacts the N terminus of the peptide, as has been shown for HLA DR and the homologous I-E molecules. PMID- 9874797 TI - Segregation of steroid receptor coactivator-1 from steroid receptors in mammary epithelium. AB - Steroid receptor coactivator-1 (SRC-1) family members interact with steroid receptors, including estrogen receptor alpha (ERalpha) and progesterone receptor (PR), to enhance ligand-dependent transcription. However, the expression of ERalpha and SRC-1 was found to be segregated in distinct subsets of cells within the epithelium of the estrogen-responsive rat mammary gland. This finding was in contrast to the finding for the stroma, where significant numbers of cells coexpressed ERalpha and SRC-1. Treatment of animals with estrogen induced PR expression in the ERalpha-expressing mammary epithelial cells in the absence of detectable SRC-1 and did not affect the segregated pattern of SRC-1 and ERalpha expression. PR was neither expressed nor induced by estrogen treatment in stroma, despite the coexpression of ERalpha and SRC-1. These results suggest that SRC-1 is not necessary for ERalpha-mediated induction of PR in mammary epithelial cells and is also not sufficient for PR induction in stromal cells expressing both ERalpha and SRC-1. Furthermore, the expression of SRC-1 in a subpopulation of mammary epithelial cells distinct from those expressing ERalpha or PR raises the possibility that SRC-1 has cell type-specific functions other than simply to act as coactivator for ERalpha or PR in the mammary epithelium. PMID- 9874798 TI - NOEY2 (ARHI), an imprinted putative tumor suppressor gene in ovarian and breast carcinomas. AB - Using differential display PCR, we have identified a gene [NOEY2, ARHI (designation by the Human Gene Nomenclature Committee)] with high homology to ras and rap that is expressed consistently in normal ovarian and breast epithelial cells but not in ovarian and breast cancers. Reexpression of NOEY2 through transfection suppresses clonogenic growth of breast and ovarian cancer cells. Growth suppression was associated with down-regulation of the cyclin D1 promoter activity and induction of p21(WAF1/CIP1). In an effort to identify mechanisms leading to NOEY2 silencing in cancer, we found that the gene is expressed monoallelically and is imprinted maternally. Loss of heterozygosity of the gene was detected in 41% of ovarian and breast cancers. In most of cancer samples with loss of heterozygosity, the nonimprinted functional allele was deleted. Thus, NOEY2 appears to be a putative imprinted tumor suppressor gene whose function is abrogated in ovarian and breast cancers. PMID- 9874799 TI - Severe cardiomyopathy in mice lacking dystrophin and MyoD. AB - The mdx mouse, a mouse model of Duchenne muscular dystrophy, carries a loss-of function mutation in dystrophin, a component of the membrane-associated dystrophin-glycoprotein complex. Unlike humans, mdx mice rarely display cardiac abnormalities and exhibit dystrophic changes only in a small number of heavily used skeletal muscle groups. By contrast, mdx:MyoD-/- mice lacking dystrophin and the skeletal muscle-specific bHLH transcription factor MyoD display a severe skeletal myopathy leading to widespread dystrophic changes in skeletal muscle and premature death around 1 year of age. The severely increased phenotype of mdx:MyoD-/- muscle is a consequence of impaired muscle regeneration caused by enhanced satellite cell self-renewal. Here we report that mdx:MyoD-/- mice developed a severe cardiac myopathy with areas of necrosis associated with hypertrophied myocytes. Moreover, heart tissue from mdx:MyoD-/- mice exhibited constitutive activation of stress-activated signaling components, similar to in vitro models of cardiac myocyte adaptation. Taken together, these results support the hypothesis that the progression of skeletal muscle damage is a significant contributing factor leading to development of cardiomyopathy. PMID- 9874800 TI - Decrease in telomerase activity in U-87MG human glioblastomas after treatment with an antagonist of growth hormone-releasing hormone. AB - Antagonists of growth hormone-releasing hormone (GH-RH) inhibit the growth of various tumors through mechanisms that involve the suppression of the insulin like growth factor I and/or insulin-like growth factor II levels or secretion. In the present study, we tested the hypothesis that the tumor inhibition is associated with a decrease in telomerase activity because telomerase is considered obligatory for continued tumor growth. Nude mice bearing xenografts of U-87MG human glioblastomas were treated with GH-RH antagonist MZ-5-156. Telomerase activity was assessed by the telomerase repeat amplification protocol. Treatment with MZ-5-156 reduced levels of telomerase activity as compared with controls. When U-87 glioblastomas, H-69 small cell lung carcinomas, H-23 non small cell lung carcinomas, and MDA-MB-468 breast carcinoma cells were cultured in vitro, addition of 3 microM MZ-5-156 also inhibited telomerase activity. Reverse transcription-PCR analysis revealed that in U-87MG glioblastomas, the expression of the hTRT gene encoding for the telomerase catalytic subunit was significantly decreased by MZ-5-156, whereas the levels of mRNA for hTR and TP1, which encode for the telomerase RNA and telomerase-associated protein, respectively, were unaffected. The repression of the telomerase activity was not accompanied by a significant decrease of mRNA level for the c-myc protooncogene that regulates telomerase. Our findings suggest that tumor inhibition induced by the GH-RH antagonists in U-87MG glioblastomas is associated with the down regulation of the hTRT gene, resulting in a decrease in telomerase activity. Further studies are needed to establish whether GH-RH antagonists produce telomerase inhibition in other tumors. PMID- 9874801 TI - Virus-specific immunity after gene therapy in a murine model of severe combined immunodeficiency. AB - Human severe combined immunodeficiency (SCID) can be caused by defects in Janus kinase 3 (JAK3)-dependent cytokine signaling pathways. As a result, patients are at high risk of life-threatening infection. A JAK3 -/- SCID mouse model for the human disease has been used to test whether transplant with retrovirally transduced bone marrow (BM) cells (JAK3 BMT) could restore immunity to an influenza A virus. The immune responses also were compared directly with those for mice transplanted with wild-type BM (+/+ BMT). After infection, approximately 90% of the JAK3 BMT or +/+ BMT mice survived, whereas all of the JAK3 -/- mice died within 29 days. Normal levels of influenza-specific IgG were present in plasma from JAK3 BMT mice at 14 days after respiratory challenge, indicating restoration of B cell function. Influenza-specific CD4(+) and CD8(+) T cells were detected in the spleen and lymph nodes, and virus-specific CD8(+) effectors localized to the lungs of the JAK3 BMT mice. The kinetics of the specific host response correlated with complete clearance of the virus within 2 weeks of the initial exposure. By contrast, the JAK3 -/- mice did not show any evidence of viral immunity and were unable to control this viral pneumonia. Retroviral mediated JAK3 gene transfer thus restores diverse aspects of cellular and humoral immunity and has obvious potential for human autologous BMT. PMID- 9874802 TI - Toward a cancer therapy with boron-rich oligomeric phosphate diesters that target the cell nucleus. AB - The viability of boron neutron capture therapy depends on the development of tumor-targeting agents that contain large numbers of boron-10 (10B) atoms and are readily taken up by cells. Here we report on the selective uptake of homogeneous fluorescein-labeled nido-carboranyl oligomeric phosphate diesters (nido-OPDs) by the cell nucleus and their long-term retention after their delivery into the cytoplasm of TC7 cells by microinjection. All nido-OPDs accumulated in the cell nucleus within 2 h after microinjection. However, nido-OPDs in which the carborane cage was located on a side chain attached to the oligomeric backbone were redistributed between both the cytoplasm and nucleus after 24 h of incubation, whereas nido-OPDs in which the carborane cage was located along the oligomeric backbone remained primarily in the nucleus. Furthermore, cell-free incubation of digitonin-permeabilized TC7 cells with the nido-OPDs resulted in nuclear accumulation of the compounds, thus corroborating the microinjection studies. Our observation of fluorescence primarily located in the cell nucleus indicates that nuclear-specific uptake of sufficient amounts of 10B for effective boron neutron capture therapy ( approximately 10(8)-10(9) 10B atoms/tumor cell) via nido-OPDs is achievable. PMID- 9874803 TI - A natural variant of the cysteine protease virulence factor of group A Streptococcus with an arginine-glycine-aspartic acid (RGD) motif preferentially binds human integrins alphavbeta3 and alphaIIbbeta3. AB - The human pathogenic bacterium group A Streptococcus produces an extracellular cysteine protease [streptococcal pyrogenic exotoxin B (SpeB)] that is a critical virulence factor for invasive disease episodes. Sequence analysis of the speB gene from 200 group A Streptococcus isolates collected worldwide identified three main mature SpeB (mSpeB) variants. One of these variants (mSpeB2) contains an Arg Gly-Asp (RGD) sequence, a tripeptide motif that is commonly recognized by integrin receptors. mSpeB2 is made by all isolates of the unusually virulent serotype M1 and several other geographically widespread clones that frequently cause invasive infections. Only the mSpeB2 variant bound to transfected cells expressing integrin alphavbeta3 (also known as the vitronectin receptor) or alphaIIbbeta3 (platelet glycoprotein IIb-IIIa), and binding was blocked by a mAb that recognizes the streptococcal protease RGD motif region. In addition, mSpeB2 bound purified platelet integrin alphaIIbbeta3. Defined beta3 mutants that are altered for fibrinogen binding were defective for SpeB binding. Synthetic peptides with the mSpeB2 RGD motif, but not the RSD sequence present in other mSpeB variants, blocked binding of mSpeB2 to transfected cells expressing alphavbeta3 and caused detachment of cultured human umbilical vein endothelial cells. The results (i) identify a Gram-positive virulence factor that directly binds integrins, (ii) identify naturally occurring variants of a documented Gram positive virulence factor with biomedically relevant differences in their interactions with host cells, and (iii) add to the theme that subtle natural variation in microbial virulence factor structure alters the character of host pathogen interactions. PMID- 9874804 TI - The voltage sensitive Lc-type Ca2+ channel is functionally coupled to the exocytotic machinery. AB - Although N- and P-type Ca2+ channels predominant in fast-secreting systems, Lc type Ca2+ channels (C-class) can play a similar role in certain secretory cells and synapses. For example, in retinal bipolar cells, Ca2+ entry through the Lc channels triggers ultrafast exocytosis, and in pancreatic beta-cells, evoked secretion is highly sensitive to Ca2+. These findings suggest that a rapidly release pool of vesicles colocalizes with the Ca2+ channels to allow high Ca2+ concentration and a tight coupling of the Lc channels at the release site. In binding studies, we show that the Lc channel is physically associated with synaptotagmin (p65) and the soluble N-ethylmaleimide-sensitive attachment proteins receptors: syntaxin and synaptosomal-associated protein of 25 kDa. Soluble N-ethylmaleimide-sensitive attachent proteins receptors coexpressed in Xenopus oocytes along with the Lc channel modify the kinetic properties of the channel. The modulatory action of syntaxin can be overcome by coexpressing p65, where at a certain ratio of p65/syntaxin, the channel regains its unaltered kinetic parameters. The cytosolic region of the channel, Lc753-893, separating repeats II-III of its alpha1C subunit, interacts with p65 and "pulls" down native p65 from rat brain membranes. Lc753-893 injected into single insulin-secreting beta-cell, inhibits secretion in response to channel opening, but not in response to photolysis of caged Ca2+, nor does it affect Ca2+ current. These results suggest that Lc753-893 competes with the endogenous channel for the synaptic proteins and disrupts the spatial coupling with the secretory apparatus. The molecular organization of the Lc channel and the secretory machinery into a multiprotein complex (named excitosome) appears to be essential for an effective depolarization evoked exocytosis. PMID- 9874805 TI - Neuroprotection with noninvasive neurotrophin delivery to the brain. AB - Brain-derived neurotrophic factor (BDNF) is neuroprotective in the ischemic hippocampus if the neurotrophin is injected directly into the brain. However, the efficacy of BDNF via peripheral (i.v.) administration is limited by the lack of transport of the neurotrophin through the brain capillary wall, which makes up the blood-brain barrier (BBB) in vivo. The present studies describe a molecular reformulation of BDNF that incorporates polyethylene glycol (PEG) moieties at surface carboxyl residues, to optimize plasma pharmacokinetics, and links pegylated BDNF to the OX26 mAb, which undergoes receptor-mediated transport through the BBB via the brain capillary endothelial transferrin receptor. The BDNF-PEG 2000-biotin conjugated to OX26/streptavidin was administered i.v. daily to rats for 1 week after a 12-min period of transient forebrain ischemia. The neuronal density in the CA1 sector of the hippocampus was decreased 68 +/- 10% at 1 week after the ischemia. There was no neuroprotective effect of the unconjugated BDNF or unconjugated OX26 mAb. However, the hippocampal CA1 neuronal density was normalized by i.v. administration of the BDNF-PEG 2000-biotin conjugated to OX26/streptavidin. These studies demonstrate that peripherally administered BDNF may have neuroprotective effects in brain, if the neurotrophin is reformulated to (i) optimize plasma pharmacokinetics with carboxyl-directed pegylation, and (ii) enable transport through the BBB by coupling to brain transport vectors. PMID- 9874806 TI - Neurons in the dorsal column white matter of the spinal cord: complex neuropil in an unexpected location. AB - It is common to think of gray matter as the site of integration in neural circuits and white matter as the wires that connect different groups of neurons. The dorsal column (DC) white matter, for example, is the spinal cord axonal pathway through which a topographic map of the body is conveyed to the somatosensory cortex. We now describe a network of neurons located along the midline of the DCs. The neurons are present in several mammals, including primates and birds, and have a profuse dendritic arbor that expresses both the neuron-specific marker, microtubule-associated protein-2, and the neurokinin-1 receptor, a target of the neuropeptide, substance P. Electron microscopy and double immunostaining for synaptophysin and a marker of gamma-aminobutyric acid ergic terminals documented a rich synaptic input to these neurons. Finally, injection of a gamma-aminobutyric acid type A receptor antagonist or of substance P into the cerebrospinal fluid of the rat spinal cord induced Fos expression and internalization of the neurokinin-1 receptor in these neurons, respectively, indicating that the DC neurons are under tonic inhibitory control and can respond to neurotransmitters that circulate in the cerebrospinal fluid. PMID- 9874807 TI - Structural requirements of ligands for the oxysterol liver X receptors LXRalpha and LXRbeta. AB - LXRalpha and -beta are nuclear receptors that regulate the metabolism of several important lipids, including cholesterol and bile acids. Previously, we have proposed that LXRs regulate these pathways through their interaction with specific, naturally occurring oxysterols, including 22(R)-hydroxycholesterol, 24(S)-hydroxycholesterol, and 24(S),25-epoxycholesterol. Using a ligand binding assay that incorporates scintillation proximity technology to circumvent many of the problems associated with assaying extremely hydrophobic ligands, we now demonstrate that these oxysterols bind directly to LXRs at concentrations that occur in vivo. To characterize further the structural determinants required for potent LXR ligands, we synthesized and tested a series of related compounds for binding to LXRs and activation of transcription. These studies revealed that position-specific monooxidation of the sterol side chain is requisite for LXR high-affinity binding and activation. Enhanced binding and activation can also be achieved through the use of 24-oxo ligands that act as hydrogen bond acceptors in the side chain. In addition, introduction of an oxygen on the sterol B-ring results in a ligand with LXRalpha-subtype selectivity. These results support the hypothesis that naturally occurring oxysterols are physiological ligands for LXRs and show that a rational, structure-based approach can be used to design potent LXR ligands for pharmacologic use. PMID- 9874808 TI - Systemic biosynthesis of prostacyclin by cyclooxygenase (COX)-2: the human pharmacology of a selective inhibitor of COX-2. AB - Prostaglandins (PG) are synthesized by two isoforms of the enzyme PG G/H synthase [cyclooxygenase (COX)]. To examine selectivity of tolerated doses of an inhibitor of the inducible COX-2 in humans, we examined the effects of celecoxib on indices of COX-1-dependent platelet thromboxane (Tx) A2 and on systemic biosynthesis of prostacyclin in vivo. Volunteers received doses of 100, 400, or 800 mg of celecoxib or 800 mg of a nonselective inhibitor, ibuprofen. Ibuprofen, but not celecoxib, significantly inhibited TxA2-dependent aggregation, induced ex vivo by arachidonic acid (83 +/- 11% vs. 11. 9 +/- 2.2%; P < 0.005) and by collagen. Neither agent altered aggregation induced by thromboxane mimetic, U46619. Ibuprofen reduced serum TxB2 (-95 +/- 2% vs. -6.9 +/- 4.2%; P < 0.001) and urinary excretion of the major Tx metabolite, 11-dehydro TxB2 (-70 +/- 9.9% vs. 20.3 +/- 5.3%; P < 0.05) when compared with placebo. Despite a failure to suppress TxA2-dependant platelet aggregation, celecoxib had a modest but significant inhibitory effect on serum TxB2 4 hr after dosing. By contrast, both ibuprofen and celecoxib suppressed a biochemical index of COX-2 activity (endotoxin induced PGE2 in whole blood ex vivo) to a comparable degree (-93.3 +/- 2% vs. -83 +/- 6.1%). There was no significant difference between the doses of celecoxib on COX-2 inhibition. Celecoxib and ibuprofen suppressed urinary excretion of the prostacyclin metabolite 2,3 dinor 6-keto PGF1alpha. These data suggest that (i) platelet COX-1-dependent aggregation is not inhibited by up to 800 mg of celecoxib; (ii) comparable COX-2 inhibition is attained by celecoxib (100-800 mg) and ibuprofen (800 mg) after acute dosing; and (iii) COX-2 is a major source of systemic prostacyclin biosynthesis in healthy humans. PMID- 9874809 TI - Atrial natriuretic peptide and oxytocin induce natriuresis by release of cGMP. AB - Our hypothesis is that oxytocin (OT) causes natriuresis by activation of renal NO synthase that releases NO followed by cGMP that mediates the natriuresis. To test this hypothesis, an inhibitor of NO synthase, L-nitroarginine methyl ester (NAME), was injected into male rats. Blockade of NO release by NAME had no effect on natriuresis induced by atrial natriuretic peptide (ANP). This natriuresis presumably is caused by cGMP because ANP also activates guanylyl cyclase, which synthesizes cGMP from GTP. The 18-fold increase in sodium (Na+) excretion induced by OT (1 microgram) was accompanied by an increase in urinary cGMP and preceded by 20 min a 20-fold increase in NO3- excretion. NAME almost completely inhibited OT-induced natriuresis and increased NO3- excretion; however, when the dose of OT was increased 10-fold, a dose that markedly increases plasma ANP concentrations, NAME only partly inhibited the natriuresis. We conclude that the natriuretic action of OT is caused by a dual action: generation of NO leading to increased cGMP and at higher doses release of ANP that also releases cGMP. OT-induced natriuresis is caused mainly by decreased tubular Na+ reabsorption mediated by cGMP. In contrast to ANP that releases cGMP in the renal vessels and the tubules, OT acts on its receptors on NOergic cells demonstrated in the macula densa and proximal tubules to release cGMP that closes Na+ channels. Both ANP- and OT induced kaliuresis also appear to be mediated by cGMP. We conclude that cGMP mediates natriuresis and kaliuresis induced by both ANP and OT. PMID- 9874810 TI - Isolation of a cytokinin gene, ZOG1, encoding zeatin O-glucosyltransferase from Phaseolus lunatus. AB - Zeatin is the most active and ubiquitous of the naturally occurring cytokinins. The O-glucoside of zeatin, found in all plants examined, is considered to be important in cytokinin transport, storage, and protection against cytokinin oxidases. The enzyme UDPglucose:zeatin O-glucosyltransferase (EC 2.4.1.203) was previously isolated from Phaseolus lunatus seeds. Immunoscreening of an expression library with monospecific antibody resulted in the isolation of a cDNA encoding the enzyme. The recombinant protein efficiently converts labeled zeatin to O-glucosylzeatin and has properties similar to the native enzyme. The cDNA of 1.5 kb contains an ORF encoding a 51. 4-kDa polypeptide of 459 amino acids. The sequence is unique based on a BLAST search of data bases. The genomic sequence, isolated with PCR using specific primers based on the cDNA sequence, does not contain introns. The cloning of this gene provides the tools for further study of the regulation of cytokinin metabolism and analysis of the precise role of O glucosylzeatin in plant development. PMID- 9874811 TI - The Lateral suppressor (Ls) gene of tomato encodes a new member of the VHIID protein family. AB - The ability of the shoot apical meristem to multiply and distribute its meristematic potential through the formation of axillary meristems is essential for the diversity of forms and growth habits of higher plants. In the lateral suppressor mutant of tomato the initiation of axillary meristems is prevented, thus offering the unique opportunity to study the molecular mechanisms underlying this important function of the shoot apical meristem. We report here the isolation of the Lateral suppressor gene by positional cloning and show that the mutant phenotype is caused by a complete loss of function of a new member of the VHIID family of plant regulatory proteins. PMID- 9874812 TI - Genes controlling fertilization-independent seed development in Arabidopsis thaliana. AB - We have cloned two genes, FIS1 and FIS2, that control both fertilization independent seed development and postpollination embryo development in Arabidopsis. These genes confer female gametophytic phenotypes. FIS2 encodes a protein with a C2H2 zinc-finger motif and three putative nuclear localization signals, indicating that it is likely to be a transcription factor. FIS1 encodes a protein with homology to the Drosophila Polycomb group gene Enhancer-of-zeste and is identical to the recently described Arabidopsis gene MEDEA. FIS1 is a protein with a number of putative functional domains, including the SET domain present in Enhancer-of-zeste-related proteins. Comparison of the position of the lesions in the fis1 and medea mutant alleles indicates that fis1 is a null allele producing a truncated polypeptide lacking all the protein domains whereas the deduced protein from medea lacks only the SET domain. We present a model of the role of FIS1 and FIS2 gene products in seed development. PMID- 9874813 TI - Diversity and molecular evolution of the RPS2 resistance gene in Arabidopsis thaliana. AB - The RPS2 gene in Arabidopsis thaliana governs resistance to strains of the bacterial pathogen, Pseudomonas syringae pv. tomato, that express the avrRpt2 gene. The two loci are involved in a gene-for-gene interaction. Seventeen accessions of A. thaliana were sequenced to explore the diversity present in the coding region of the RPS2 locus. An unusually high level of nucleotide polymorphisms was found (1.26%), with nearly half of the observed polymorphisms resulting in amino acid changes in the RPS2 protein. Seven haplotypes (alleles) were identified and their evolutionary relationships deduced. Several of the alleles conferring resistance were found to be closely related, whereas susceptibility to disease was conferred by widely divergent alleles. The possibility of selection at the RPS2 locus is discussed. PMID- 9874814 TI - Mechanisms of color constancy under nearly natural viewing. AB - Color constancy is our ability to perceive constant surface colors despite changes in illumination. Although color constancy has been studied extensively, its mechanisms are still largely unknown. Three classic hypotheses are that constancy is mediated by local adaptation, by adaptation to the spatial mean of the image, or by adaptation to the most intense image region. We measure color constancy under nearly natural viewing conditions, by using a design that allows us to test these three hypotheses directly. By suitable stimulus manipulation, we are able to titrate the degree of constancy between 11% and 83%, indicating that we have achieved good laboratory control. Our results rule out all three classic hypotheses and thus suggest that there is more to constancy than can be easily explained by the action of simple visual mechanisms. PMID- 9874815 TI - Evidence of hominid-like precision grip capability in the hand of the Miocene ape Oreopithecus. AB - Functional and allometric analyses of the hand of the late Miocene ape Oreopithecus bambolii (Tuscany, Italy) reveal a series of features that reflect an improved grasping capability including firm pad-to-pad precision gripping that apes are unable to perform. Related features such as hand length, relative thumb length, a deep and large insertion area for the tendon of the long thumb flexor, and the form of the metacarpal 2/capitate articulation are not present in extant or fossil apes. In these features, the Oreopithecus hand closely matches the pattern of early hominids, presumably as a response to similar functional demands. PMID- 9874816 TI - Proposition: long-term changes in irradiated tissues are due principally to vascular damage in the tissues. PMID- 9874817 TI - Dosimetric prerequisites for routine clinical use of new low energy photon interstitial brachytherapy sources. Recommendations of the American Association of Physicists in Medicine Radiation Therapy Committee. Ad Hoc Subcommittee of the Radiation Therapy Committee. PMID- 9874818 TI - Anisotropy functions for 125I and 103Pd sources. AB - The anisotropy function was defined by AAPM Task Group 43 in a recent report, and reference data were provided. However, some of the data appeared to have large experimental errors. In this work, anisotropy functions are presented for three low-energy photon-emitting sources commonly used for brachytherapy: 125I source models 6702 and 6711, and the model 200 103Pd source. The data were generated via a two-step process: the sources' intrinsic radiation emission pattern was determined from in-air measurements, and these data were used as input to Monte Carlo calculations of the fluence distribution in water. The new anisotropy function values for iodine compare well with earlier data from diode measurements, and seem more physically realistic than some data provided by TG43. Previous palladium data measured with TLDs are more variable, but on the average are similar to the data presented here. The new data have estimated uncertainties of 3%-6%; this level of accuracy should be completely adequate for clinical calculations. PMID- 9874819 TI - Experimental determination of dosimetry functions of Ir-192 sources. AB - Experimental data related to the dosimetric characteristics of Ir-192 brachytherapy sources are limited. The aim of this work was to obtain the dosimetry functions required by the American Association of Physicists in Medicine Task Group 43 for both a low and a high dose-rate iridium-192 brachytherapy source through dose measurements in a water-equivalent phantom. Dose measurements have been performed using lithium fluoride thermoluminescent detectors positioned in a polystyrene phantom at distances from the source that vary from 1 to 10 cm, with 1 cm intervals, and at angles that vary from 0 degree to 170 degrees with 10 degrees intervals. The anisotropy functions, radial dose functions, and dose rate constants were determined for both brachytherapy sources. The precision of results obtained on those relatively fine intervals of angles and distances provides clinics with the possibility to use and interpolate the complete data sets for treatment planning. PMID- 9874820 TI - Monte Carlo dose calculations for a new ovoid shield system for carcinoma of the uterine cervix. AB - The dose distribution for an ovoid with a new tungsten shielding design was determined using Monte Carlo simulation. Standard Cesium-137 tube sources, tungsten shielding, and aluminum ovoid applicator were each modeled as a collection of solid objects. Dose was calculated in planes above, below, in front of, and on the sides of the colpostat. The Monte Carlo results were compared with the results from a parametrized calculation algorithm and good agreement was obtained. The dose distribution matrix derived from the parametrized algorithm can be used for clinical treatment planning. PMID- 9874822 TI - A new genetic algorithm technique in optimization of permanent 125I prostate implants. AB - Real time optimized treatment planning at the time of the implant is desirable for ultrasound-guided transperineal 125I permanent prostate implants. Currently available optimization algorithms are too slow to be used in the operating room. The goal of this work is to develop a robust optimization algorithm, which is suitable for such application. Three different genetic algorithms (sGA, sureGA and securGA) were developed and compared in terms of the number of function evaluations and the corresponding fitness. The optimized dose distribution was achieved by searching the best seed distribution through the minimization of a cost function. The cost function included constraints on the periphery dose of the planned target volume, the dose uniformity within the target volume, and the dose to the critical structure. Adjustment between the peripheral dose, the dose uniformity and critical structure dose can be achieved by varying the weighting factors in the cost function. All plans were evaluated in terms of the dose nonuniformity ratio, the conformation number and the dose volume histograms. Among these three GA algorithms, the securGA provided the best performance. Within 2500 function evaluations, the near optimum results were obtained. For a large target volume (5 cm x 4 cm x 4.5 cm) including urethra with 20 needles, the computer time needed for the optimization was less than 5 min on a HP735 workstation. The results showed that once the best set of parameters was found, they were applicable for all sizes of prostate volume. For a fixed needle geometry, the optimized plan showed much better dose distribution than that of nonoptimized plan. If the critical structure was considered in the optimization, the dose to the critical structure could be minimized. In the cases of irregular and skewed needle geometry, the optimized treatment plans were almost as good as ideal needle geometry. It is concluded that this new genetic algorithm (securGA) allows for an efficient and rapid optimization of dose distribution, which is suitable for real time treatment planning optimization for ultrasound-guided prostate implant. PMID- 9874821 TI - Correlation of medical dosimetry quality indicators to the local tumor control in patients with prostate cancer treated with iodine-125 interstitial implants. AB - The treatment of prostate cancer by 125I interstitial implants has been extensively studied with mixed results by one institution or another. A recent study from Hahnemann [Int. J. Radiat. Oncol., Biol., Phys. 21,955-960 (1991)] reported results that were extremely poor compared to those reported in an earlier study at Yale [Int. J. Radiat. Oncol., Biol., Phys. 14, 1153-1157 (1988)] or those in an Eastern Virginia Study [Cancer 63, 2415-2420 (1989)]; differences in 5-yr survival rates being more than a factor of 2. Such large discrepancies from institution to institution led us to a reexamination of the dosimetry. This study analyzed quantitatively three-dimensional dosimetric parameters of 110 prostate cancer patients treated with 125I interstitial implants. The study searched for "cutoff" values in each parameter that divided the patients into two groups with statistically significant differences in the local recurrence-free survival rates. A comparison of the three-dimensional isodose surfaces of patients with favorable values in all of the parameters to those patients with all unfavorable parameters show how these characteristics translated into poor dose coverage and much inhomogeneity within the implant even for cases that met the traditional criteria for adequacy (160 Gy to the tumor volume). Patients in the favorable group had 10-yr survival rates higher by a factor of up to 2 compared to those in the unfavorable group. The strong correlation of three dimensional volume-dose parameters to the local control rate observed in this study further emphasizes how important it is to assess the three-dimensional dosimetric adequacy of interstitial implants before deciding on their clinical efficacy. If implants are performed with appropriate attention to dosimetry parameters, excellent clinical results are obtained. On the other hand, if dosimetry parameters are not correct, the implant results can be poor. PMID- 9874824 TI - The radiation induced magnetic resonance image intensity change provides a more efficient three-dimensional dose measurement in MRI-Fricke-agarose gel dosimetry. AB - A detailed methodology has been developed to map the spatial dose distribution in a Fricke-agarose gel based on the radiation induced image intensity change in the gel's magnetic resonance (MR) images. Besides the linear correlation between the change in the gel's spin-lattice relaxation rate and the absorbed dose, it is shown here that the radiation induced image intensity change for T1-weighted spin echo images with TE << TR correlates exponentially to the absorbed dose. Furthermore, at the lower dose region (< 15 Gy), the correlation is fairly linear and its sensitivity is high. The minimum detectable dose is shown to be equivalent to the one obtained using the conventional R1-based approach. Since only one T1-weighted image is required for the dose evaluation, compared to the R1-based method, the total MR imaging time can be reduced from hours to a few minutes. This extensive time reduction avoids ferric ion diffusion effects and provides a practical way to simply and effectively measure the three-dimensional dose distribution using the Fricke-agarose dosimeter gel. PMID- 9874823 TI - Magnetic resonance imaging of microbubbles in a superheated emulsion chamber for brachytherapy dosimetry. AB - This paper describes development of magnetic resonance imaging (MRI) techniques for three-dimensional (3D) imaging of a position-sensitive detector for brachytherapy dosimetry. The detector is a 0.5 l chamber containing an emulsion of halocarbon-115 droplets in a tissue-equivalent glycerin-based gel. The halocarbon droplets are highly superheated and expand into vapor microbubbles upon irradiation. Brachytherapy sources can be inserted into the superheated emulsion chamber to create distributions of bubbles. Three-dimensional MRI of the chamber is then performed. A 3D gradient-echo technique was optimized for spatial resolution and contrast between bubbles and gel. Susceptibility gradients at the interfaces between bubbles and gel are exploited to enhance contrast so microscopic bubbles can be imaged using relatively large voxel sizes. Three dimensional gradient-echo images are obtained with an isotropic resolution of 300 microns over a 77 mm x 77 mm x 9.6 mm field-of-view in an imaging time of 14 min. A post-processing technique was developed to semi-automatically segment the bubbles from the images and to assess dose distributions based on the measured bubble densities. Relative dose distributions are computed from MR images for a 125I brachytherapy source and the results compare favorably to relative radial dose distributions calculated as recommended by Task Group 43 of the American Association of Physicists in Medicine. PMID- 9874827 TI - Three-dimensional optimization of treatment planning for gamma unit treatment system. AB - During a treatment using the Leksell gamma unit, the physician and physicist need to determine a treatment plan by changing the parameters such as collimator sizes, the position of isocenters and isocenters' weights. This is a complex problem because the set of parameters is large, especially when targets are geometrically close to a critical structure. For this reason, we present here an optimization algorithm, namely the multiplier penalty method, to mathematically determine those parameters. Two cases are presented in this article: the first one is really planned by a physicist in a clinical treatment, and is redone in our optimization algorithm to show the effectiveness of this method; the second one is theoretical where a critical structure is placed close to the target volume. The results show that this method achieves an excellent conformation to the specified isodose curve with the contour of the target volume, allowing minimal damage to surrounding healthy tissue. PMID- 9874825 TI - Rapid portal imaging with a high-efficiency, large field-of-view detector. AB - The design, construction, and performance evaluation of an electronic portal imaging device (EPID) are described. The EPID has the same imaging geometry as the current mirror-based systems except for the x-ray detection stage, where a two-dimensional (2D) array of 1 cm thick CsI(Tl) detector elements are utilized. The approximately 18% x-ray quantum efficiency of the scintillation detector and its 30 x 40 cm2 field-of-view at the isocenter are greater than other area imaging EPIDs. The imaging issues addressed are theoretical and measured signal to-noise ratio, linearity of the imaging chain, influence of frame-summing on image quality and image calibration. Portal images of test objects and a humanoid phantom are used to measure the performance of the system. An image quality similar to the current devices is achieved but with a lower dose. With approximately 1 cGy dose delivered by a 6 MV beam, a 2 mm diam structure of 1.3% contrast and an 18 mm diam object of 0.125% contrast can be resolved without using image-enhancement methods. A spatial resolution of about 2 mm at the isocenter is demonstrated. The capability of the system to perform fast sequential imaging, synchronized with the radiation pulses, makes it suitable for patient motion studies and verification of intensity-modulated beams as well as its application in cone-beam megavoltage computed tomography. PMID- 9874828 TI - Beam characteristics and clinical possibilities of a new compact treatment unit design combining narrow pencil beam scanning and segmental multileaf collimation. AB - Not until the last decade has flexible intensity modulated three-dimensional dose delivery techniques with photon beams become a clinical reality, first in the form of heavy metal transmission blocks and other beam compensators, then in dynamic and segmented multileaf collimation, and most recently by scanning high energy narrow electron and photon beams. The merits of various treatment unit and bremsstrahlung target designs for high-energy photon therapy are investigated theoretically for two clinically relevant target sites, a cervix and a larynx cancer both in late stages. With an optimized bremsstrahlung target it is possible to generate photon beams with a half-width of about 3 cm at a source to axis distance (SAD) of 100 cm and an initial electron energy of 50 MeV. By making a more compact treatment head and shortening the SAD, it is possible to reduce the half-width even further to about 2 cm at a SAD of 70 cm and still have sufficient clearance between the collimator head and the patient. One advantage of a reduced SAD is that the divergence of the beam for a given field size on the patient is increased, and thus the exit dose is lowered by as much as 1%/cm of the patient cross section. A second advantage of a reduced SAD is that the electron beam on the patient surface will be only about 8 mm wide and very suitable for precision spot beam scanning. It may also be possible to reduce the beamwidth further by increasing the electron energy up to about 60 MeV to get a photon beam of around 15 mm half-width and an electron beam as narrow as 5 mm. The compact machine will be more efficient and easy to work with, due to the small gantry and the reduced isocentric height. For a given target volume and optimally selected static multileaf collimator, it is no surprise that the narrowest possible scanned elementary bremsstrahlung beam generates the best possible treatment outcome. In fact, by delivering a few static field segments with individually optimized scan patterns, it is possible to combine the advantage of being able to fine tune the fluence distribution by the scanning system with the steeper dose gradients that can be delivered by a few static multileaf collimator segments. It is demonstrated that in most cases a few collimator segments are sufficient and often a single segment per beam portal may suffice when narrow scanned photon beams are employed, and they can be delivered sequentially with a negligible time delay. A further advantage is the increase of therapeutically useful photons and improved patient protection, since the pencil beam is only scanned where the leaf collimator is open. Consequently, some of the problems associated with dynamic multileaf collimation such as the tongue and groove and edge leakage effects are significantly reduced. Fast scanning beam techniques combined with good treatment verification systems allow interesting future possibilities to counteract patient and internal organ motions in real time. PMID- 9874826 TI - On absorbed dose in narrow 60Co gamma-ray beams and dosimetry of the gamma knife. AB - Using separate analytical functions describing primary dose, P0(dm,r), collimator scatter, Sc(r), and phantom scatter, TAR(d,r), an expression for absorbed dose in narrow 60Co gamma-ray beams is developed and each function is quantified: D(d,r) = P0(dm,r) Sc(r) TAR(d,r). The absorbed dose is calculated in beams as narrow as 0.2 cm in radius. Analytical and experimental results are compared using measured dose data for the Gamma Knife. Close agreement with experimental data is observed. PMID- 9874829 TI - Monte Carlo method to study the proton fluence for treatment planning. AB - The proton beam at the Hahn Meitner Institute (HMI) in Berlin will be used for proton therapy of eye melanoma in the near future. As part of the pre-therapeutic studies, Monte Carlo calculations have been performed to investigate the primary fluence distribution of the proton beam including the influence of scattering foils, range shifters, modulator wheels, and collimators. Any material in the beam path will modify the therapeutic beam because of energy loss, multiple scattering, range straggling, and nuclear reactions. The primary fluence information is a pre-requisite for most pencil-beam treatment planning algorithms. The measured beam penumbra has been used as one of the parameters to characterize a proton beam for further calculations in a treatment planning algorithm. However, this phenomenological quantity represents only indirect information about the properties of the proton beam. In this work, an alternative parameterization of the beam exiting the vacuum window of the accelerator, as well as the beam right in front of the patient collimator, is introduced. A beam is fully characterized if one knows (for instance from Monte Carlo simulations) the particle distribution in energy, position, and angle, i.e., the phase space distribution. Therefore, parameters derived from this distribution can provide an alternative input in treatment planning algorithms. In addition, the method of calculation is introduced as a tool to investigate the influence of modifications in the beam delivery system on the behavior of the therapeutic proton beam. PMID- 9874830 TI - Study of dosimetry consistency for kilovoltage x-ray beams. AB - In this paper, the consistency of kilovoltage (tube potentials between 40 and 300 kV) x-ray beam dosimetry using the "in-air" method and the in-phantom measurement has been studied. The procedures for the measurement of the central-axis depth dose curve, which serve as a link between the dose at the reference depth to the dose elsewhere in a phantom, were examined. The uncertainties on the measured dose distributions were analyzed with the emphasis on the surface dose measurement. The Monte Carlo method was used to calculate the perturbation correction factors for a photon diode and a NACP plane-parallel ionization chamber at different depths in a water phantom irradiated by 100-300 kV (2.43 mm Al-3.67 mm Cu half-value layer) x-ray beams. The depth-dose curves measured with these two detectors, after correcting for the perturbation effect (up to 15% corrections), agreed with each other to within 1.5%. Comparisons of the doses at the phantom surface and at 2 cm depth in water for photon beams of 100-300 kV tube potential obtained using the "backscatter" method and those using the "in phantom" measurement have shown that the "in-air" method can be equally applied to this energy range if the depth-dose curve can be measured accurately. To this end, measured depth ionization curves require depth-dependent correction factors. PMID- 9874831 TI - A room-based diagnostic imaging system for measurement of patient setup. AB - A room-based diagnostic x-ray imaging system for routine measurement of radiotherapy patient orientation has been developed. The system consists of a pair of room-mounted x-ray tubes and a portable imager consisting of an orthogonal pair of phosphor screens, a mirror/lens system, a CCD camera, and computer software for comparing images of the patient to reference images. Orthogonal pairs of images can be acquired quickly and with relatively little exposure, allowing correction of patient setup on a daily basis. This could limit patient setup error to the uncertainty in the measurement and repositioning processes, a potentially significant improvement over the present standard. PMID- 9874832 TI - Fetal dose estimates for electron-beam treatment to the chest wall of a pregnant patient. AB - The purpose of this report is to discuss dosimetry and shielding for electron beam radiotherapy of pregnant patients. Specifically, we have determined fetal dose for a pregnant patient considering electron radiotherapy for a chest wall recurrence of breast cancer. The treatment was simulated using an anthropomorphic phantom, and the measured dose to the unshielded fetus for this plan was 5.3 cGy, a level at which risk to the fetus is uncertain. Therefore abdominal shielding, consisting of 6.6 cm of lead, was used to reduce the dose to the fetus to less than 1.5 cGy, a level considered to be of little risk. We further found that using the lower (instead of upper) variable trimmer bars to define the field edge closest to the fetus resulted in approximately 30% lower dose to the fetus. These results show that it is possible to reduce fetal dose to acceptable limits in electron-beam radiotherapy of the chest wall using the general principles recommended for photon-beam radiotherapy. PMID- 9874833 TI - Intracavitary ultrasound phased arrays for prostate thermal therapies: MRI compatibility and in vivo testing. AB - A 62 element MRI-compatible linear phased array was designed and constructed to investigate the feasibility of using transrectal ultrasound for the thermal therapeutic treatment of prostate cancer and benign prostatic hyperplasia. An aperiodic design technique developed in a previous study was used in the design of this array, which resulted in reduced grating lobe levels by using an optimized random distribution of unequally sized elements. The element sizes used in this array were selected to be favorable for both grating lobe levels as determined by array aperiodicity and array efficiency as determined by width to thickness ratios. The heating capabilities and MRI compatibility of the array were tested with in vivo rabbit thigh muscle heating experiments using MRI temperature monitoring. The array produced therapeutic temperature elevations in vivo at depths of 3-6 cm and axial locations up to 3 cm off the central axis and increased the size of the heated volume with electronic scanning of a single focus. The ability of this array to be used for ultrasound surgery was demonstrated by creating necrosed tissue lesions in vivo using short high-power sonications. The ability of the array to be used for hyperthermia was demonstrated by inducing therapeutic temperature elevations for longer exposures. Based on the acoustic and heating performance of this array, it has the potential to be clinically useful for delivering thermal therapies to the prostate and other target volumes close to body cavities. PMID- 9874834 TI - Air-cooling of direct-coupled ultrasound applicators for interstitial hyperthermia and thermal coagulation. AB - The feasibility of using air-cooling to improve the thermal penetration of direct coupled interstitial ultrasound (US) applicators was investigated using biothermal simulations, bench experiments, phantom testing, and in vivo thermal dosimetry. Two applicator configurations using tubular US transducers were constructed and tested. The first design, intended for simultaneous thermobrachy therapy, utilizes a 2.5 mm OD transducer with a central lumen to accommodate a radiation source from remote afterloaders. The second applicator consists of a 2.2 mm OD transducer designed for coagulative thermal therapy. Both designs provide cooling of the inner transducer surface by the counterflow of chilled air or CO2 gas through the annulus of the enclosed applicator. The average convective heat transfer (ha) associated with each applicator was determined empirically from curve-fits of radial steady-state temperatures measured in a tissue mimicking phantom. High levels of convective heat transfer (ha > 500 W m-2 degrees C-1) were demonstrated in both designs at relatively low flow rates (< 5 L min-1). Transient and steady-state radial heating profiles were also measured in vivo (pig thigh muscle) with and without cooling. The therapeutic radius for hyperthermia (41-45 degrees C) was extended from 5-6 mm (without cooling) to 11 19 mm with air-cooling (4.8 L min-1, airflow 10 degrees C), effectively doubling and tripling the thermal penetration in vivo. Similar improvements were demonstrated at higher temperatures with the thermal coagulation applicator. Biothermal simulations, which modeled the physical, thermal, and acoustic parameters of the air-cooled applicator and surrounding tissue, were also used to investigate potential improvements in heating patterns. The simulated radial heating profiles with transducer cooling demonstrated significantly enhanced thermal penetration over the experimental range of convective transfer, and also agreed with in vivo results. These theoretical and experimental results clearly show air-cooling controls the transducer surface temperature, significantly increases thermal penetration, and produces a greater treatment volume for direct coupled US applicators in hyperthermia and thermal coagulation. PMID- 9874835 TI - Establishing minimum performance standards, calibration intervals, and optimal exposure values for a whole breast digital mammography unit. AB - Methods are developed to establish minimum performance standards, calibration intervals, and criteria for exposure control for a whole breast digital mammography system. A prototype phantom was designed, and an automatic method programmed, to analyze CNR, resolution, and dynamic range between CCD components in the image receptor and over time. The phantom was imaged over a 5 month period and the results are analyzed to predict future performance. White field recalibration was analyzed by subtracting white fields obtained at different intervals. Exposure effects were compared by imaging the prototype phantom at different kVp, filtration (Mo vs Rh) and mAs. Calcification detection tests showed that phantom images, obtained at 28 kVp with a Mo/Mo anode/filter and low mAs technique, often could not depict Al2O3 specks 0.24 mm in diameter, while a 28 kVp Mo/Rh, higher mAs technique usually could. Stability of the system tested suggests that monthly phantom imaging may suffice. Differences in CCD performance are greater (12%) than differences in a single CCD over time (6%). White field recalibration is needed weekly because of pixel variations in sensitivity which occur if longer intervals between recalibration occur. When mean glandular dose is matched, Rh filtration gives better phantom performance at 28 kVp than Mo filtration at 26 kVp and is recommended for clinical exposures. An aluminum step wedge shows markedly increased dynamic range when exit exposure is increased by using a higher energy spectrum beam. Phantoms for digital mammography units should cover the entire image receptor, should test intersections between components of the receptor, and should be automatically analyzed. PMID- 9874836 TI - Electrocardiogram-correlated image reconstruction from subsecond spiral computed tomography scans of the heart. AB - Subsecond computed tomography (CT) scanning offers potential for improved heart imaging. We therefore developed and validated dedicated reconstruction algorithms for imaging the heart with subsecond spiral CT utilizing electrocardiogram (ECG) information. We modified spiral CT z-interpolation algorithms on a subsecond spiral CT scanner. Two new classes of algorithms were investigated: (a) 180 degrees CI (cardio interpolation), a piecewise linear interpolation between adjacent spiral data segments belonging to the same heart phase where segments are selected by correlation with the simultaneously recorded ECG signal and (b) 180 degrees CD (cardio delta), a partial scan reconstruction of 180 degrees + delta with delta < fan angle, resulting in reduced effective scan times of less than 0.5 s. Computer simulations as well as processing of clinical data collected with 0.75 s scan time were carried out to evaluate these new approaches. Both 180 degrees CI and 180 degrees CD provided significant improvements in image quality. Motion artifacts in the reconstructed images were largely reduced as compared to standard spiral reconstructions; in particular, coronary calcifications were delineated more sharply and multiplanar reformations showed improved contiguity. However, new artifacts in the image plane are introduced, mostly due to the combination of different data segments. ECG-oriented image reconstructions improve the quality of heart imaging with spiral CT significantly. Image quality and the display of coronary calcification appear adequate to assess coronary calcium measurements with conventional subsecond spiral CT. PMID- 9874837 TI - On segmentation of lung parenchyma in quantitative computed tomography of the lung. AB - Our purpose in this study was to investigate the influence of segmentation threshold and number of erosions on parameters used in quantitative computed tomography (CT) of the lung (erosions are shrink operations on the segmented area). Parameters assessed were mean lung density, area of the segmented lung, two percentiles, and the pixel index, which is the relative area of the histogram below -905 Hounsfield Units (HU). We analyzed images of ten emphysematous and ten nonemphysematous patients, that had been scanned at carina level in inspiration and expiration, using sections of 1, 2, 3, 5, and 10 mm in combination with a standard, a smooth, and an ultrasmooth reconstruction kernel. The lungs were segmented using pixel tracing at thresholds of -200, -400, and -600 HU with 0-4 erosions, followed by histogram analysis. The area of the segmented lungs decreased with 0.9%-3.2% per 100 HU decrease in threshold and with 2.2%-3.1% per erosion, dependent on patient group and respiratory status. Estimated mean lung density changed up to 30% by changing the threshold and the number of erosions. The pixel index and the 10th percentile depended only slightly on threshold and number of erosions, but the 90th percentile showed a strong dependence of up to 40%. It is concluded that the segmentation protocol can have a large impact on densitometric parameters and that standardization is mandatory for obtaining comparable results. Ideally a threshold equal to the average of the densities of lung and soft tissue should be used, but -400 HU will do in a limited but common density range (-910 to -790 HU). For densitometry two erosions are recommended, for volumetry zero erosions should be used. PMID- 9874838 TI - The effect of phosphor persistence on image quality in digital x-ray scanning systems. AB - A digital x-ray scanning system offers several advantages over conventional film screen systems. However, there are sources of image degradation resulting from the scanning motion, such as motion blur due to the temporal response of the phosphor. This mechanism produces an asymmetrical blur, requiring the use of the complex optical transfer function (OTF) rather than the normal modulation transfer function (MTF) for correct characterization of image resolution. The luminescence response of eight phosphors was measured under pulsed x-ray excitation. A weighted exponential model was used to represent the primary luminescence. The dominant luminescence life-times ranged from 2.7 microseconds for Gd2O2S:Pr to 558 microseconds for Gd2O2S:Tb. The long term response was also measured, monitoring significant increases in a slow form of luminescence known as afterglow. Afterglow was modeled by an inverse power law equation. Afterglow was found to be strong in two of the phosphors studied (ZnCdS:Ag and YTaO4). In selecting a phosphor for a scanning system, it must satisfy several criteria, including a fast temporal response. Thus, a phosphor like Gd2O2S:Tb, which has a slow luminescence, but otherwise excellent imaging properties, may not be as useful as a more rapid phosphor like CsI:Tl. PMID- 9874839 TI - Characteristics of metal-plate/film detectors at therapy energies. I. Modulation transfer function. AB - Measurements of modulation transfer function (MTF) for front and back metal plate/film portal detectors are reported for the Cobalt-60 and 10 MV spectra. The detectors consist of a double-emulsion portal film secured between plates of Al, Cu, brass, or Pb with thicknesses varying from 0 to 4.81 mm. Secondary electrons produced within the front plate generate the main signal, but the MTF decreases with an increase in front plate thickness greater than the maximum range of electrons Rmax because of photon scatter in the front plate. Because the decrease of MTF with backplate thickness ceases for backplate thickness greater than Rmax, the MTF is influenced more by the backscatter electrons than the backscatter photons. PMID- 9874840 TI - Characteristics of metal-plate/film detectors at therapy energies. II. Detective quantum efficiency. AB - Noise power spectrum (NPS) and detective quantum efficiency (DQE) for metal plate/film portal detectors are reported for the Co-60 and Linac 10 MV spectra. The detectors consist of a double-emulsion portal film secured between plates of aluminum, copper, brass, or lead. The NPS was found to be independent of the detector sensitivity demonstrating that the film grain noise dominates over the quantum noise. Although the detector signal increases with density of the backplates, the resultant electron-backscatter increases detector blur, thus decreasing the DQE. The lowest DQEs are therefore produced with the lead backplates, irrespective of the front plates. For a given backplate and for front plate thicknesses less than the maximum electron range, the DQE increases with density of the front plate. PMID- 9874841 TI - A multiresolution restoration method for cardiac SPECT imaging. AB - In this study we present a multiresolution based method for restoring cardiac SPECT projections. Original projections were decomposed into a set of sub-band frequency images by using analyzing functions localized in both the space and frequency domain. This representation allows a simple denoising and restoration procedure by discarding high-frequency channels and performing inversion only in low frequencies. The method was evaluated in bull's eye reconstructions of a realistic cardiac chest phantom with a custom-made liver insert and 99mTc liver to-heart activity ratios (LHAR) of 0:1, 1.5:1, 2.5:1, and 3.5:1. The cardiac phantom in free air was used as the reference standard. Reconstructions were performed by filtered backprojection using (1) no correction; (2) restoration without attenuation correction; (3) attenuation correction without restoration; and (4) restoration and attenuation correction. The attenuation correction was carried out with the Chang's method for one iteration. Results were compared with those obtained using an optimized prereconstruction Metz filter. Quantitative analysis was performed by calculating the normalized chi-square measure and mean +/- s.d. of bull's eye counts. In reconstructions with high liver activity (LHAR > 2), attenuation correction without restoration severely distorted the polar maps due to the spill-over of liver activity into the inferior myocardial wall. Both restoration methods when combined with an attenuation correction compensated this artifact and yielded uniform polar maps similar to that of the standard reference. There was no visual or quantitative difference between the performance of Metz filtering and multiresolution restoration. However, the main advantage of the multiresolution method is that it states a more concise and straightforward approach to the restoration problem. Multiresolution based methods does not require information about the object image or optimization processes, such as in conventional nuclear medicine restoration filters. In addition, the method is easy to implement using DFT techniques and potentially can be extended to noniterative spatially shift-invariant restorations in SPECT. PMID- 9874842 TI - Comment on "Proposition: radiation hormesis should be elevated to a position of scientific respectability" [Med. Phys. 25, 1407-1410 (1998)]. PMID- 9874843 TI - Comment on "Dosimetry of interstitial brachytherapy sources: recommendations of the AAPM Radiation Therapy Committee Task Group No. 43" [Med. Phys. 22, 209-234 (1995)]. PMID- 9874844 TI - Comment on "Cerebral tumor volume calculations using planimetric and eigenimage analysis" [Med. Phys. 23, 2035-2042 (1996)]. PMID- 9874845 TI - Predicting prognosis in patients with superficial bladder cancer. AB - Bladder cancer is the most common urologic malignancy and is expected to affect approximately 54,000 people in 1998. Superficial bladder tumors (Tis, Ta, and T1 lesions) account for approximately 70% to 80% of these malignancies. Although many advances have been made in the management of patients with superficial bladder cancer, such disease often recurs and can progress, leading to death. It is therefore imperative to find an accurate method of identifying patients at risk for disease progression. Many recent investigations have been conducted to determine whether new biological markers will help predict disease progression and, to a lesser extent, tumor response to treatment. These new markers include DNA ploidy, S-phase, certain monoclonal antibodies, the p53 (alias TP53) tumor suppressor gene, the retinoblastoma (Rb) gene, cell adhesion molecules, and angiogenesis. It is hoped that such prognostic indicators, coupled with cytoscopic and pathologic characteristics of the tumor, will lead to selective aggressive treatment of patients at high risk for progression while sparing low risk patients from unnecessary procedures. PMID- 9874846 TI - Monoclonal antibody approved for metastatic breast cancer. PMID- 9874848 TI - Clinical trials referral resource. Clinical trials in gastric cancer. PMID- 9874847 TI - Lower genital tract neoplasia in women with HIV infection. AB - Women who are infected with human immunodeficiency virus (HIV) are at greater risk for the development of lower genital tract neoplasia than are HIV-negative women. Among HIV-positive women, those who are more severely immunosuppressed appear to be at higher risk for cervical intraepithelial neoplasia (CIN), also known as squamous intraepithelial lesions (SILs). Women who are HIV-positive also are more likely than HIV-negative women to have multifocal lower genital tract neoplasia. Cervical cancer is one of the most important acquired immune deficiency syndrome (AIDS)--related malignancies in women. Cancer and intraepithelial neoplasia of the lower genital tract can be persistent, progressive, recurrent, and difficult to treat in HIV-positive women. The most effective method for treating SILs has not been determined. Regular performance of Pap smears in HIV-positive women is of critical importance, as is careful examination of the entire lower genital tract. Also, women with high-grade intraepithelial or cervical cancer should be tested for HIV. PMID- 9874849 TI - Clinical status and optimal use of rituximab for B-cell lymphomas. AB - Rituximab (IDEC-C2B8 [Rituxan]) is a chimeric anti-CD20 monoclonal antibody (MoAb) that was recently approved by the FDA for the treatment of patients with low-grade or follicular B-cell non-Hodgkin's lymphoma. Its potential efficacy in other B-cell malignancies is currently being explored. This article reviews the mechanisms of action of rituximab, as well as preclinical data and results of the clinical trials that led to its approval. Also discussed are the mechanics of administering rituximab on the recommended weekly x 4 outpatient schedule. Finally, the article describes ongoing and planned trials of rituximab in other dosage schedules, in other B-cell neoplasms, and in conjunction with chemotherapy. As the first MoAb to gain FDA approval for the treatment of a malignancy, rituximab signals the beginning of a promising new era in cancer therapy. PMID- 9874850 TI - Cancer chemoprevention. Part 2: Hormones, nonclassic antioxidant natural agents, NSAIDs, and other agents. AB - This two-part series provides an up-to-date summary of the various chemopreventive agents currently in development and testing. Part 1, published in last month's issue, focused on the retinoids, such as all-trans-retinoic acid (ATRA [Vesanoid]), 9-cis-retinoic acid (9cRA), and isotretinoin (Accutane), and the carotenoids, such as beta-carotene and lycopene, and other "classic" antioxidants, such as vitamins E and C and selenium. This second part centers on hormonally mediated chemopreventives, such as tamoxifen (Nolvadex), finasteride (Proscar), oral contraceptives, and dehydroepiandrosterone (DHEA). A number of these agents have site-specific mechanisms of action and will be discussed as such. Also discussed in part 2 are various nonclassic antioxidant natural agents, including calcium, the polyphenols, the isothiocyanates, and genistein; nonsteroidal anti-inflammatory drugs (NSAIDS), such as aspirin and sulindac; difluoromethylornithine (DFMO [Eflornithine]); oltipraz; and N-acetylcysteine. PMID- 9874851 TI - Neurofibromatosis 1 in childhood. AB - Neurofibromatosis 1 (NF1) is an autosomal-dominant disorder with an incidence of approximately 1 in 3,000. The cardinal features of the disorder are cafe au lait spots, axillary freckling, cutaneous neurofibromas, and iris hamartomas (Lisch nodules). Common complications include learning disability, scoliosis, and optic gliomas. Other complications (e.g., peripheral nerve malignancy, bony deformities, and epilepsy) are individually rare. The mainstay of care for patients with NF1 is anticipatory guidance, and the early detection and symptomatic treatment of disease complications. Counseling of patients and their families should provide a realistic overview of possible disease complications, while emphasizing that most individuals with NF1 lead healthy and productive lives. The gene for NF1 has been identified, and future cell biology research will focus on understanding the pathogenetic mechanisms that underly the diverse manifestations of the disorder. PMID- 9874852 TI - The diagnosis and management of neurofibromatosis 2 in childhood. AB - Neurofibromatosis 2 (NF2) is an autosomal-dominant condition that causes multiple benign nervous system tumors, especially vestibular schwannoma. Although frequently confused with the more common neurofibromatosis 1, NF2 presents a distinct set of diagnostic, genetic, and management issues. The presentation and natural history of NF2 differs in children and adults, with eighth nerve dysfunction often overshadowed by the effects of other tumors on the nervous system. Molecular diagnostics is an important tool for early recognition of NF2, and when performed in the context of appropriate counseling and follow-up, may lead to a better final outcome of the disease. Further research into the molecular genetic etiology of NF2 promises to broaden the possibilities of therapy for this devastating disorder. PMID- 9874853 TI - Diagnosis and management of tuberous sclerosis complex. AB - Tuberous sclerosis complex (TSC) is an autosomal-dominant neurocutaneous disorder with a high spontaneous mutation rate. Understanding of this disorder has greatly increased in recent years. Two chromosomal loci can produce the TSC phenotype: 9q34 and 16p13. These appear to code for proteins that have a tumor suppressor function. TSC results in hamartomas that affect various organ systems, most commonly brain, skin, heart, and kidney. Previously thought to consist of intractable seizures, facial angiofibromas, and dementia, increasing numbers of persons with less severe involvement have been identified. Diagnostic criteria, various types of lesions, and medical management are reviewed. PMID- 9874854 TI - Renal angiomyolipomas, cysts, and cancer in tuberous sclerosis complex. AB - Tuberous sclerosis is a multisystem syndrome characterized by neurological symptoms and tumors in multiple organs, including kidney, brain, skin, eyes, heart, and lung. The kidney and brain are the two most frequently affected organs in TSC, and renal disease is a leading cause of death in TSC patients. Three types of tumors occur in TSC kidneys: (1) angiomyolipomas, which are benign tumors composed of smooth muscle, fat, and vessels; (2) epithelial cysts; and (3) malignant tumors. This review focuses on the clinical, pathological, and molecular features of these tumors. PMID- 9874855 TI - Parallels between tuberous sclerosis complex and neurofibromatosis 1: common threads in the same tapestry. AB - Neurofibromatosis type 1 (NF1) and tuberous sclerosis complex (TSC) represent two neurocutaneous disorders in which affected individuals develop tumors at an increased frequency. Although the clinical manifestations of these disorders are distinctive, the identification of the genes responsible for these disorders has demonstrated remarkable similarities on a molecular level between the NF1 and TSC tumor suppressor gene products. The NF1 and TSC2 gene products are hypothesized to function as growth regulators by modulating the activities of small GTPase molecules. The overlap between the functions of these tumor suppressor genes has yielded important insights into the molecular pathogenesis underlying each of these disorders and suggested possible pharmacological therapies specifically targeted for affected individuals. PMID- 9874856 TI - Ataxia telangiectasia. AB - The cloning of ATM in 1995, the gene responsible for ataxia-telangiectasia, opened a dimension of biological research that is as complex and intriguing to cell biologists as this classic disorder has been to clinicians for four decades. The phenotype is both variable and stereotyped, with significant differences between patients in the rate of progression or appearance of the multiple features yet consistent in their characteristic nature. Ataxia telangiectasia usually has been misdiagnosed for the first few years of life, while accurate diagnosis might most impact family planning. Newly produced ATM-deficient transgenic mice express most of the cellular features of the disorder but have yet to mimic the distinctive neurodegeneration. PMID- 9874857 TI - Intrathecal opioids provide satisfactory conditions in labor analgesia and spinal anesthesia for cesarean section. PMID- 9874858 TI - Effects of dobutamine, norepinephrine and epinephrine on intramucosal pH and hemodynamics of dogs during endotoxic shock. AB - This study assessed the effects of dobutamine (DOB), epinephrine (EPI) and norepinephrine (NE) on gastric tissue oxygenation indicated by gastric intramucosal pH (pHi) and hemodynamics in dogs subjected to endotoxic shock. Twenty-four dogs were assigned to four groups of 6 dogs each: endotoxin without catecholamine and endotoxin with DOB, or EPI or NE. Endotoxic shock was induced by intravenous injection of 3 mg/kg of E. coli over 1 min, with an additional 3 mg/kg over the next 2 hrs. Dogs were resuscitated with normal saline to maintain pulmonary capillary wedge pressure (PCWP) near baseline levels. Catecholamines were infused at 0.1, 0.4 and 1.6 micrograms/kg/min (EPI and NE) and 2.5, 5.0 and 10.0 micrograms/kg/min (DOB) for 30 min at each rate. After 2 hrs of endotoxemia, mean arterial pressure (MAP) and cardiac index (CI) and oxygenation delivery index (DO2I) for all dogs decreased by 46.5%, 43.9% and 15.1% respectively, while pHi decreased from 7.47 to 7.10. Endotoxemia increased blood lactate by 142%. Following fluid resuscitation, EPI (1.6 micrograms/kg/min) further increased lactate by 178% (1.22 to 3.4 mmol/L). No correlation was found between tonometry pHi and lactate (R2 = 0.003), pHi and pHa (R2 = 0.231), pHi and DO2I (R2 = 0.056) nor between intramucosal PCO2 and PaCO2 (R2 = 0.005). pHi did not reflect the improvements in cardiovascular hemodynamics observed following administration of catecholamines. NE improved MAP, CI and DO2I whereas DOB produced similar effects as NE but further reduced SVR. EPI produced similar effects as NE. DOB, NE and EPI further decreased pHi. EPI significantly (P < 0.05) increased blood lactate levels more than DOB and NE. PMID- 9874859 TI - NitroG-L-arginine methyl ester decreases minimum alveolar concentration of isoflurane and reduces brain nitric oxide synthase activity in rats. AB - BACKGROUND: Nitric Oxide (NO), an endogenous messenger produced by the enzyme nitric oxide synthase (NOS), is recently introduced to be involved in inhalational anesthesia. We have previously reported that a specific NOS inhibitor, nitroG-L-arginine methyl ester (L-NAME), reduces the value of minimum alveolar concentration (MAC) for isoflurane anesthesia in rabbits. The purpose of this study is to evaluate the effects of the NOS inhibitor, L-NAME, on isoflurane MAC and NOS activity in rats. METHODS: Adult Wistar rats receiving isoflurane inhalation were randomly divided into two groups, with eight rats in each group. In the study group, L-NAME 30 mg/kg was given 60 min before the inhalation of isoflurane. Normal saline was given to the control group instead. The data of MAC, blood pressure (BP), and heart rate (HR) were recorded. The vital signs, such as EtCO2, PaO2, and temperature, were maintained within normal ranges. The activity of NOS in cerebellum was assessed by measuring the conversion of L-[3H] arginine to L-[3H] citrulline. All data were presented as mean +/- SD. Statistical analysis was performed using Student's t-test, where P < 0.05 was considered significant. RESULTS: In the presence of L-NAME (30 mg/kg), the MAC for isoflurane was markedly reduced from 1.6 +/- 0.20% (study group) to 1.0 +/- 0.09% (control group) (P < 0.05). The activity of cNOS in cerebellum was 220.09 +/- 23.64 (pmol/mg protein/30 min) in the control group, and in contrast a sharp reduction as low as to 115.40 +/- 24.85 (pmol/mg protein/30 min) was seen in the study group. CONCLUSIONS: The involvement of NO in the mechanism of isoflurane anesthesia can be demonstrated by the fact that the NOS inhibitor, L-NAME reduces the level of MAC and the cerebral NOS activity in rats. PMID- 9874860 TI - Intradermal injection of tramadol has local anesthetic effect: a comparison with lidocaine. AB - BACKGROUND: We observed that intravenous retention of tramadol with a pneumatic tourniquet on the arm inflated to 70 mmHg for one minute could effectively reduce the subsequent propofol injection pain. Tramadol is a central-acting analgesic. The local analgesic effect of tramadol on reducing propofol injection pain is not well known. METHODS: To explore this problem we conducted a double-blind study on intradermal injections of tramadol 25 mg, lidocaine 5 mg and normal saline (all in 0.5 ml volume) which were given to each of the 10 healthy volunteers on the forearm at random. Pain on injections and the degree of local analgesia to pinprick, light touch and cold at each injection site were scored on a 0-4 scale at designated intervals. RESULTS: 5% tramadol, similar to 1% lidocaine, rendered loss of sensation to pin prick, light touch and cold for 30 min after intradermal injection as compared with normal saline (p < 0.01). CONCLUSIONS: We concluded that intradermal injection of tramadol or lidocaine can produce local anesthetic effect. PMID- 9874861 TI - Pre-anesthetic oral clonidine is effective to prevent post-spinal shivering. AB - BACKGROUND: Shivering is a common event during spinal anesthesia. Customarily we just treat it rather than prevent it. This study was designed to evaluate the efficacy of oral clonidine as a premedication to prevent post-spinal shivering. METHODS: One hundred males of ASA physical status I-III, aged above 40, scheduled for elective urological surgery under spinal anesthesia, were included in this study. All participants were randomly divided into the clonidine and control groups. They received either oral clonidine 150 micrograms (n = 48) or placebo (n = 52) 90 min before spinal anesthesia in a double-blind fashion. Spinal blockade was induced with heavy bupivacaine to a dermatomal level near T10. The shivering was graded as: none, no perceptible tension of muscles observed; mild, slight muscle tonus (masseter muscle); moderate, real shivering (proximal muscles); and severe, generalized shivering (whole body). The tympanic membrane temperature was recorded 30 min after spinal anesthesia. Data were expressed as mean +/- standard deviation. Chi-square and Student's t-test were used. A p value less than 0.05 was considered statistically significant. RESULTS: The incidence of post-spinal shivering, which was graded as none, mild, moderate, and severe, showed statistically significant differences (p < 0.05) between clonidine 150 micrograms and placebo (83% vs. 42%, 10% vs. 6%, 10% vs. 19%, 0% vs. 33%, respectively) during the 30 min immediately after spinal anesthesia. The respective mean tympanic temperature in oral clonidine and placebo groups showed no difference (clonidine vs. control = 35.9 +/- 0.8 degrees C vs. 35.9 +/- 0.7 degrees C). CONCLUSIONS: Pre-anesthetic medication with oral clonidine 150 micrograms is effective to prevent post-spinal shivering in patients undergoing elective urological surgery. PMID- 9874862 TI - The maternal and fetal effects of the addition of sufentanil to 0.5% spinal bupivacaine for cesarean delivery. AB - BACKGROUND: Opioid added to local anesthetic for spinal anesthesia was first introduced into clinical practice in 1979 with intrathecal morphine as a forerunner. As morphine is water soluble and has prolonged action, late respiratory depression following spinal anesthesia is not infrequent and is the most serious complication that causes our concern. Sufentanil which is more hydrophobic than morphine also with shorter duration of action and quicker onset when injected into the subarchnoid space could be more effective and a safer drug as an adjuvant to local anesthetic in spinal anesthesia. METHODS: Forty-one parturients who had given consent to spinal anesthesia for Cesarean delivery, were anesthetized with 12.5 mg of 0.5% bupivacaine alone or in combination with 10 micrograms sufentanil in a randomized double blind manner. They were assigned either to C group (Control group) in which nothing is added to the local anesthetic and S group (Study group) in which sufentanil was added to the local anesthetic. RESULTS: Perioperatively, hypotension occurred more in S group (17 against 11) but chest discomfort was less (3 against 7). Within 3 h after anesthesia 3 out of 19 parturients in S group requested analgesics but almost all parturients in C group did so. CONCLUSIONS: The addition of intrathecal sufentanil to 0.5% bupivacaine for spinal anesthesia improved perioperative discomfort and significantly reduced the demand of post-operative analgesia but on the other hands, it tended to increase perioperative hypotension and cause mild pruritus. PMID- 9874863 TI - The effect of coagulation protection with combination of epsilon aminocaproic acid and plasma saver in open-heart surgery. AB - BACKGROUND: Bleeding remains a major complication and a major determinant in the prognosis of open-heart surgery. Coagulopathy related to cardiopulmonary bypass (CPB) seems to be the culprit. Since homologous blood transfusion in many occasions is not only responsible for mobidity and mortality but also increases medical costs. Therefore, the application of autologous blood transfusion including components such as PRBC, FFP and platelets concentrate is inevitable and comes in its stead. To reduce the use of homologous plasma and platelets transfusion in open-heart surgery, we designed a study to utilize the combination of autologous platelet rich plasma (PRP) and epsilon aminocaproic acid (EACA) to evaluate its effects on blood loss and blood component transfusion in open-heart patients. METHODS: Sixty patients who received elective cardiac surgery were randomly divided into 3 groups: 1. Control group; 2. EACA group (150 mg/kg, i.v. before CPB); 3. PRP-EACA group (PRP 10 ml/kg harvested with a plasma saver followed by i.v. EACA 150 mg/kg). Anesthesia was uniform in all patients. Coagulation profile was evaluated by thromboelastography (TEG) during the operation. Blood loss during operation and the amount of drainage from the chest tubes in the postoperative period were recorded and compared between groups. RESULTS: Patients who were given EACA injection before CPB saw less blood loss perioperatively and received less transfusion of blood components. TEG analysis showed that patients who received EACA injection had a better coagulation profile and the platelet function was also better after CPB. However, no additive effect can be attained from combination of autologous PRP transfusion and EACA injection. CONCLUSIONS: With Pre-CPB EACA as protection, reduction of both blood loss and blood transfusion could be realized in open-heart surgery. PMID- 9874864 TI - Repeated lumbar sympathetic blockade for complex regional pain syndromes type I- a case report. AB - The causal mechanisms of reflex sympathetic dystrophy are not limited purely to sympathetic hyperactivity, but sympathetic blockade and physical therapy are still the mainstays of current treatment modalities. We reported a case of reflex sympathetic dystrophy over the left lower leg, who was successfully treated with repeated lumbar sympathetic blocks and physical therapy. PMID- 9874865 TI - Anesthetic management of a parturient with Eisenmenger's syndrome and preeclampsia during cesarean section--a case report. AB - It has been recommended that women with Eisenmenger's syndrome (ES) are better not to become pregnant and pregnancy may justifiably be terminated by artificial abortion to avoid high maternal mortality and coherent fetal mortality. We present a case report about a parturient with ventricular septal defect (VSD) and ES who received general anesthesia for Cesarean section (C/S) because of preeclampsia, as a result of which she finally succumbed to an episode of intraoperative hypotension in spite of vigorous cardiopulmonary resuscitation. The death was thought to be precipitated by continuous deterioration of maternal health during the 3rd trimester of gestation. The anesthetic management of pregnant ES patients in confinement was reviewed and discussed, and the possible etiological factors relevant to the tragic outcome were also explored. PMID- 9874866 TI - Succinylcholine-induced cardiac arrest in unsuspected becker muscular dystrophy- a case report. AB - A five year-old boy undergoing elective tonsillectomy sustained cardiac arrest following the administration of a single dose of succinylcholine during induction of anesthesia. With a 10-minute cardiopulmonary resuscitation (CPR) during which intravenous calcium gluconate, epinephrine, and sodium bicarbonate were given and DC counter shock applied, we were successful to restore cardiac activity without neurological sequelae. The cause of cardiac arrest we speculated was hyperkalemia, possibly secondary to succinylcholine-induced rhabdomyolysis. It is suggested that succinylcholine should not be used in patients with known or suspected muscular dystrophy. PMID- 9874867 TI - Unexpected pheochromocytoma--a case report of anesthesia in a uremic patient. AB - Pheochromocytoma is an uncommon neoplasm causing blood pressure changes. It may go undiagnosed in uremic patients in whom hypertension is common. The preferential diagnostic work-up, including urine and serum catecholamine measurements, is unsuitable for uremic patients due to anuria and inherent increases in serum catecholamine levels. Here, we present a case of uremia, scheduled for right adrenalectomy and simple nephrectomy, who had been sustaining hypertension. Malignant hypertension was only discovered during surgical manipulation of the adrenal tumor. Pheochromocytoma was highly suspected. The patient was treated with nitroglycerin, fentanyl, and increased concentration of isoflurane to deepen the anesthesia. However, the results were only fair and after ligation of the tumor veins, hypotension ensued. Pathological examination verified the diagnosis of pheochromocytoma. In reviewal of the whole course of treatment, it was considered that the placement of a pulmonary artery catheter to detect volume shifts and changes in hemodynamic status was indicated. Related papers were reviewed and application of diagnostic tools and their limitations, safety and efficacy of anesthetics and vasodilators, and possible complications relevant to management of pheochromocytoma were also discussed. In conclusion, it is important to remain alert, administer agents cautiously, and set up complete monitoring if needed, and carry out postoperative intensive care to lessen complications in the anesthetic management of this rare and dangerous incidentaloma encountered during anesthesia. PMID- 9874868 TI - An unexpected complication using the LMA and nasogastric tube. PMID- 9874869 TI - GA--end of an era. PMID- 9874871 TI - Research in general practice. PMID- 9874870 TI - Biting the bullet--report of the Dental Auxiliaries Review Group. PMID- 9874872 TI - Research in general practice. PMID- 9874873 TI - The registered dental nurse. PMID- 9874874 TI - Amalgam fillings during pregnancy. PMID- 9874875 TI - Care required when prescribing carbamazepine. PMID- 9874876 TI - Orthopantomographic detection of a metallic foreign body in the neck. PMID- 9874877 TI - Orthodontic auxiliaries. PMID- 9874878 TI - Poor standards of dental prosthetics. PMID- 9874879 TI - Dental receptionist referrals. PMID- 9874880 TI - Electronic file transfer protocols. PMID- 9874881 TI - Using an aspirating syringe for regional block anaesthesia. PMID- 9874882 TI - Focus on loupes. AB - This article aims to introduce the reader to the use of magnification in clinical dentistry and describes the lens systems available. The main principles of dental loupes are discussed and some of the potential problems outlined in order to help clinicians choose a magnification suited to their needs, provide years of service, and reduce the risk of eye, neck and back strain. The additional benefits and disadvantages of adding supplementary illumination, and the options available, are also considered. PMID- 9874883 TI - Efficient use of the Internet. AB - This section explains how to make most of your time on-line by providing tips on how to reduce the cost of using the internet. Suggestions are given on how to get the best from your existing hardware and software. There are also recommendations for some additional hardware and software. PMID- 9874884 TI - Briefing paper: xylitol, caries and plaque. AB - Xylitol is a naturally occurring sweetener. Beyond being non-cariogenic, it exhibits both passive and active anti-caries properties and is used in confectionery and oral care products. PMID- 9874885 TI - The prevalence and natural history of periodontal disease in Britain from prehistoric to modern times. AB - OBJECTIVE: To investigate the prevalence of periodontitis in British populations during the past 3000 years. To discuss the relevance of these findings to modern populations denied access to modern dental treatment. DESIGN: Observation and assessment of periodontal status of skeletal material using a recently developed method of assessment. SETTING: Archaeological material from various collections throughout UK covering prehistoric, mediaeval and seventeenth to nineteenth century. SUBJECTS: 504 individuals. MAIN OUTCOME MEASURES: The periodontal status of individual septa was recorded as to whether they were healthy, had gingivitis or periodontitis. The prevalence of periodontitis among individuals and populations was assessed. RESULTS: The prevalence of periodontitis appears to have remained virtually constant during the past 3000 years in Britain, despite considerable changes in the oral environment. CONCLUSIONS: The significance of these findings in respect of untreated populations in underdeveloped countries today is considerable. It is considered essential that clinical and field studies record and report on oral environmental factors that may mimic or, by themselves, be responsible for periodontal attachment loss. PMID- 9874886 TI - Oral PowderJect: a novel system for administering local anaesthetic to the oral mucosa. AB - OBJECTIVE: To assess the feasibility of using an Oral PowderJect (OPJ) to safely deliver a dose of dry powdered anaesthetic to the oral mucosa, producing an analgesic effect. DESIGN: Single centre: Part 1. An open, non-randomised safety study to check for mucosal damage; Part 2. A double blind sham controlled study to test the anaesthetic effect. SETTING: General practice. SUBJECTS: Adult, healthy volunteers (4 male, 10 female). MATERIALS AND METHODS: Part 1. An OPJ was used to deliver powdered lidocaine hydrochloride to the mucosal surface which was then checked visually for damage. Part 2. An OPJ containing lidocaine hydrochloride (active) or an empty OPJ (sham) was fired at the oral mucosa. The treated area and an untreated (control) site were probed with the back end of a dental needle. RESULTS: The OPJ delivery caused no visible mucosal damage. The median VAS score for pain on blunt probing was 10 for the OPJ active sites. This was significantly lower than the median VAS score for the sham sites at 30 (P = 0.0033) and the control sites at 58 (P < 0.0001). CONCLUSIONS: The OPJ can safely deliver powdered lidocaine hydrochloride to the oral mucosa without causing tissue damage. The OPJ delivery of powdered lidocaine hydrochloride can significantly reduce the pain from a blunt needle probe at 1 minute post delivery. PMID- 9874887 TI - Dental practitioners' experiences on the usefulness of restorative materials in Finland 1992-1996. AB - OBJECTIVE: To analyse general dentists' choice of restorative materials in Finland, where the use of amalgam has significantly decreased and the use of composites increased during the past 10 years. DESIGN AND SETTING: Postal survey in 1997 to a random sample of general dental practitioners mainly working with adult patients. MAIN OUTCOME MEASURES: Percentage distribution of dentists' opinions. RESULTS: The response rate was 82%. Tooth-coloured restorations were considered to be more time consuming, technically complicated and to have shorter life spans. Most dentists were not willing to return to the wide-scale use of amalgam restorations. CONCLUSIONS: The rapid changes in the restorative material selection have not resulted in serious problems in restorative therapy. PMID- 9874888 TI - Case report: an autopsy study of a rare T-cell lymphoma clinically misdiagnosed as fulminant hepatic disease. AB - We describe a case of a 73-year-old male with a rare T-cell lymphoma that presented deceptively as progressive hepatic failure with fever, weight loss, pancytopenia, mental confusion, splenomegaly, and no lymphadenopathy. An alcoholic history supported the diagnosis of cirrhosis, but a liver biopsy was not performed. A bone marrow biopsy was considered unremarkable. Death occurred after a course of four months. Postmortem examination showed hepatic, splenic, lymph node, and marrow infiltration by characteristically sparse, isolated, bizarre, medium-to-large sized neoplastic cells with extensive hepatic centrilobular necrosis, steatosis, and predominant splenic involvement. Immunohistochemical markers indicated a T-cell lymphoma consistent with either an alpha/beta peripheral T-cell lymphoma or a gamma/delta lymphoma. Definitive immunotyping was not available. However, the pathologic features are most consistent with a gamma/delta T-cell lymphoma. This case is an example of a rare, rapidly progressive lymphoma, which is a recognized clinical entity, easily missed, and treatable. Its diagnostic consideration must be explicitly communicated to pathologists, because the isolated or sparse tumor cells in a lymph node, liver, or bone marrow biopsy may easily be mistaken for variants of megakaryocytes or histiocytes. PMID- 9874889 TI - Factors inhibiting use of the pneumococcal polysaccharide vaccine: a survey of Connecticut physicians. AB - BACKGROUND: The pneumococcal polysaccharide vaccine (PPV) is effective in preventing invasive pneumococcal disease, but remains underutilized. Prior surveys of physicians revealed concern regarding the safety and efficacy of the vaccine, but there has been little information published in the last 10 years that sheds light on why the vaccine remains underutilized. Although there is currently emphasis on providing PPV to hospitalized patients, there is even less known about what factors prevent PPV use in the hospital setting and chronic care setting. We performed a survey of physicians in Connecticut to determine what factors prevent utilization of the vaccine in three patient care settings. METHODS: A survey of internists and family practitioners in Connecticut that ascertained their frequency of utilization of PPV and what factors inhibited utilization of PPV. RESULTS: Three hundred ninety-seven responses are included in the analysis. Forgetting to administer the vaccine (59% of respondents) and patient refusal (55% of respondents) were the factors most frequently noted as being important in preventing vaccination in the outpatient setting. In the inpatient and chronic care settings, difficulty in determining the patient's vaccine status was also noted. Concerns regarding the efficacy or safety of the vaccine did not seem to be important. The factor that correlated most closely with the respondents' reported frequency of vaccine use was forgetting to vaccinate. CONCLUSIONS: Physicians, although accepting the efficacy of PPV, are inhibited from its more frequent use by several factors. PMID- 9874891 TI - The role of the primary pediatrician when a child dies. PMID- 9874890 TI - Antimicrobial switch therapy. PMID- 9874892 TI - In response to The ethics of organ transplantation. PMID- 9874893 TI - In response to "Pain control following elective gastrointestinal surgery: is epidural anesthesia warranted?". PMID- 9874894 TI - Humanitarian action in protracted crisis: an overview of the debates and dilemmas. PMID- 9874895 TI - Proliferating principles; or how to sup with the devil without getting eaten. AB - The international humanitarian system is seen by many to be in crisis. The reasons for this are complex but include the perception held by many that there is an increasing ambivalence on the part of powerful states to invest military, political and financial resources in upholding humanitarian principles, and a growing feeling that much humanitarian action both ignores human rights issues and can prolong conflict. As a result, much attention has focused on the notion of 'humanitarian principles' and there has been a proliferation of statements and initiatives on this topic in recent years. PMID- 9874896 TI - The death of humanitarianism?: an anatomy of the attack. AB - Over recent years, the humanitarian community has been under increasing scrutiny and attracted harsh criticism. This mounting critique of humanitarianism has shifted from being concerned primarily with the poor functioning of the humanitarian system to one targeted on basic humanitarian values. This paper is concerned to understand the factors driving the attack on humanitarian values. It identifies four apparently disparate groups who are interested in attacking these values: the anti-imperialists, the realpolitikers, the developmentalists and the neo-peaceniks. It concludes that unless humanitarian actors are aware of these diverse threats to their values and operations, they risk being co-opted or marginalised. PMID- 9874897 TI - Aid and violence, with special reference to Sierra Leone. AB - This paper looks at how departures from humanitarian principles can be accommodated, legitimised and obscured within the international humanitarian system. It looks particularly at the case of Sierra Leone between 1991 and 1995. It analyses how misinformation about the causes and dynamics of violence and regarding the aid system contribute to the erosion of humanitarian principles. PMID- 9874898 TI - Normalising the crisis in Africa. AB - Developmental relief has become the central doctrine of 'good practice' in humanitarian responses to complex political emergencies. This is despite the fact that a proliferation of such emergencies reflects a failure of development for people in those countries in crisis. Drawing on case study material from Sudan, Somalia, Rwanda and Uganda, this paper challenges assumptions made about the efficacy of developmental relief models in complex emergencies. The trend towards developmental relief practices coincides with an increasing acceptance of higher levels of humanitarian distress in Africa. Myths of aid dependency and the pursuit of sustainable programming in the midst of war are linked to a global reduction in aid. The mantra of 'local solutions to local problems' locates the causes of crises firmly within those societies in crisis. It provides a premise for international disengagement, and the denial of international responsibility for the genesis and prolongation of humanitarian crises in Africa. Assigning solutions to the poor, the marginalised and victimised through enhanced 'participation' and local financing of services sustains a myth that development is occurring, when in fact levels of distress are rising. PMID- 9874899 TI - Military intervention and humanitarian action: trends and issues. AB - An important trend in military doctrine for so-called 'peace support operations' has been to place them on a spectrum that includes coercion and enforcement. This paper focuses on British writers of doctrine as those responsible for driving the debate forward and forging a consensus among leading military powers. Their discourse is combat oriented, a fact reinforced by a trend towards strategic subcontracting to coalitions of the militarily willing and able. At the same time, there has been a move to institutionalise the involvement of military forces in relief, peace building and development activities. The overall emphasis is on stability and security to facilitate the delivery of humanitarian relief and to establish the conditions for peace-building processes. In the case of Bosnia, this involves conditionality and economic leverage. Although there is a long-established record of peace-keeping forces engaging in goodwill activities (with mixed results), the current trends contain contradictions that seem likely to contribute to the widely perceived erosion of classical humanitarian principles. PMID- 9874900 TI - In defence of humanitarianism. AB - The humanitarian crisis which followed in the wake of the genocidal regime in Rwanda in 1994 generated massive media attention and an unprecedented outpouring of international public and private assistance. In late 1997, the Rwanda refugee population in Zaire was subjected to a disaster of similarly epic proportions as a result of military action. Yet this crisis went relatively under-reported and failed to attract substantial aid funds, particularly from official donors. This paper seeks to document and account for the demise of the humanitarian imperative. It confronts a number of the criticisms of humanitarian action, concluding that, rather than being flawed, traditional humanitarian values remain valid and should be defended wherever there are situations of conflict. PMID- 9874901 TI - Children's predisaster functioning as a predictor of posttraumatic stress following Hurricane Andrew. AB - This study examined (a) children's predisaster behavioral and academic functioning as a predictor of posttraumatic stress (PTS) following Hurricane Andrew and (b) whether children who were exposed to the disaster would display a worsening of prior functioning. Fifteen months before the disaster, 92 4th through 6th graders provided self-reports of anxiety; peers and teachers rated behavior problems (anxiety, inattention, and conduct) and academic skills. Measures were repeated 3 months postdisaster; children also reported PTS symptoms and hurricane-related experiences (i.e., exposure). PTS symptoms were again assessed 7 months postdisaster. At 3 months postdisaster, children's exposure to the disaster, as well as predisaster ratings of anxiety, inattention, and academic skills, predicted PTS symptoms. By 7 months, only exposure, African American ethnicity, and predisaster anxiety predicted PTS. Prior anxiety levels also worsened as a result of exposure to the disaster. The findings have implications for identifying and treating children at risk for stress reactions following a catastrophic disaster. PMID- 9874902 TI - The role of parental anxiety in the treatment of childhood anxiety. AB - Sixty-seven children aged 7 to 14 who met diagnostic criteria for an anxiety disorder were assigned to conditions according to parental anxiety level. Within these conditions, children were randomly assigned to 1 of 2 treatments: child focused cognitive-behavioral therapy (CBT) or child-focused CBT plus parental anxiety management (CBT + PAM). At posttreatment, results indicated that within the child-anxiety-only condition, 82% of the children in the CBT condition no longer met criteria for an anxiety disorder compared with 80% in the CBT + PAM condition. Within the child + parental anxiety condition, 39% in the CBT condition no longer met criteria compared with 77% in the CBT + PAM condition. At follow-up, these differences were maintained, with some weakening over time. Results were not consistent across outcome measures. The interpretation and potential clinical implications of these findings are discussed. PMID- 9874904 TI - Utility of psychophysiological measurement in the diagnosis of posttraumatic stress disorder: results from a Department of Veterans Affairs Cooperative Study. AB - This multisite study tested the ability of psychophysiological responding to predict posttraumatic stress disorder (PTSD) diagnosis (current, lifetime, or never) in a large sample of male Vietnam veterans. Predictor variables for a logistic regression equation were drawn from a challenge task involving scenes of combat. The equation was tested and cross-validated demonstrating correct classification of approximately 2/3 of the current and never PTSD participants. Results replicate the finding of heightened psychophysiological responding to trauma-related cues by individuals with current PTSD, as well as differences in a variety of other domains between groups with and without the disorder. Follow-up analyses indicate that veterans with current PTSD who do not react physiologically to the challenge task manifest less reexperiencing symptoms, depression, and guilt. Discussion addresses the value of psychophysiological measures for assessment of PTSD. PMID- 9874903 TI - Psychophysiologic assessment of women with posttraumatic stress disorder resulting from childhood sexual abuse. AB - Heart rate, skin conductance, and left lateral frontalis and corrugator facial electromyogram responses were measured during script-driven imagery of personal childhood sexual abuse (CSA) and other life experiences among women with and without Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev., American Psychiatric Association, 1987)--diagnosed posttraumatic stress disorder (PTSD) resulting from CSA. Women with current PTSD (n = 29) showed larger physiologic responses than those who never had PTSD (n = 18) during personal sexual abuse imagery but not during imagery of stressful, nonabuse-related life experiences. Responses of individuals with lifetime, but not current, PTSD (n = 24) fell between the other groups. An a priori discriminant function, derived from physiologic responses of previously studied individuals, correctly classified 66% of women with current PTSD and 78% of women who never had PTSD. PMID- 9874905 TI - When is less treatment better? The role of social anxiety in matching methadone patients to psychosocial treatments. AB - In response to a need to match drug users to the most appropriate and cost effective level of care, it was hypothesized the socially anxious methadone maintained patients would attain greater benefit from coping skills training provided in the context of a low-intensity enhanced standard methadone maintenance intervention (E-STD) than in the context of a high-intensity, socially demanding day treatment program (DTP). Social anxiety was assessed in 307 methadone-maintained patients using the Social Anxiety and Distress Scale prior to randomization to either E-STD or DTP. The hypothesis was supported: Socially anxious patients were drug free longer during treatment, were more likely to be abstinent at treatment completion, and had greater reductions in HIV risk behaviors if assigned to the lower intensity intervention, which was provided at 1/3 the cost of the DTP. PMID- 9874906 TI - The severity of major depression and choice of treatment in primary care practice. AB - The Agency for Health Care Policy and Research Depression Guideline Panel recommended pharmacotherapy as the 1st-line treatment for more severely depressed primary care patients, but research supporting its recommendation has not been conducted with this population. A post hoc analysis was conducted, therefore, with data gathered in a randomized controlled trial about the relationship between initial level of depressive severity and functional ability, treatment with nortriptyline hydrochloride (NT) or interpersonal psychotherapy (IPT), and clinical course over 8 months among primary care patients experiencing major depression. Treatment type was unrelated to clinical course among more severely depressed patients (baseline 17-item Hamilton Rating Scale for Depression [HRSD] score > or = 20). However, less severely depressed patients (baseline 17-item HRSD score < or = 19) who were prescribed NT improved significantly more rapidly during the initial 3 months of treatment than patients provided with IPT. PMID- 9874907 TI - Dynamic systems theory as a paradigm for the study of change in psychotherapy: an application to cognitive therapy for depression. AB - Dynamic systems theory provides a conceptual framework for the study of change in psychotherapy that is consistent with that used in other sciences. A dynamic systems model of change was proposed and evaluated in the context of cognitive therapy for depression. Consistent with this model, less client protection and more destabilization of depressive patterns predicted more improvement at the end of treatment. Less protection was associated with more therapist support/stabilization. More destabilization was associated with more affective intensity in the session and with more of a therapist focus on the historical antecedents of current problems, exposure to multiple sources of corrective information, and repeated practice of new skills. Although preliminary, this pattern of findings is consistent with the model proposed and with principles of dynamic systems from other sciences. PMID- 9874908 TI - A comparison of normal forgetting, psychopathology, and information-processing models of reported amnesia for recent sexual trauma. AB - This study assessed memories for sexual trauma in a nontreatment-seeking sample of recent rape victims and considered competing explanations for failed recall. Participants were 92 female rape victims assessed within 2 weeks of the rape; 62 were also assessed 3 months postassault. Memory deficits for parts of the rape were common 2 weeks postassault (37%) but improved over the 3-month window studied (16% still partially amnesic). Hypotheses evaluated competing models of explanation that may account for reported recall deficits. Results are most consistent with information-processing models of traumatic memory. PMID- 9874909 TI - "A grief observed": the experience of HIV-related illness and death among women in a clinic-based sample in New Haven, Connecticut. AB - This study examined how women in a clinic-based sample in New Haven, Connecticut, (N = 168), have been affected by the AIDS epidemic. The aims of this study were to (a) document the proportion of women who knew individuals who were HIV positive, who were symptomatic with AIDS, or who had died from AIDS; (b) compare the demographics of women who knew someone infected with HIV with those of women who did not know anyone infected with HIV; and (c) examine prospectively the effects of the number of AIDS-related losses on women's mental health. Many women have been deeply affected by the AIDS epidemic: Nearly 3/4 of the women in this study knew at least 1 person who had died of AIDS. Women who experienced multiple AIDS-related losses over the course of the study were significantly more anxious than those who experienced no loss or 1 loss. Implications for clinical interventions are discussed. PMID- 9874910 TI - Psychosocial factors associated with the stages of change for condom use among women at risk for HIV and STDs: implications for intervention development. AB - This study examined the prevalence of consistent condom use among inner-city women at risk for HIV, measured the distribution of these women across the stages of change for condom use, determined psychosocial factors associated with the stages, and suggested intervention strategies based on the results. The 5-city sample of women aged 15-34 years consisted predominantly of African Americans. Only 18% reported consistent condom use with main partners and 45% with other partners. Logistic regressions compared women in each stage of change with those in higher stages for each partner type. Results indicated that women who practice or intend to practice consistent condom use were more likely to talk with others about condoms, acknowledge the advantages of condoms, have higher self-efficacy for condom use, and indicate that people important to them favored condom use. Intervention approaches are suggested for women in different stages of change for condom use. PMID- 9874912 TI - The prospective relationships between smoking and weight in a young, biracial cohort: the Coronary Artery Risk Development in Young Adults Study. AB - This study examined the relationship between smoking status and weight change from baseline to Year 7 in a large biracial cohort, the Coronary Artery Risk Development in Young Adults study. Unadjusted for covariates, only male smokers weighed less than nonsmokers, with no effect among women. Adjusted for covariates, male and female smokers weighed less than nonsmokers at baseline, adjusted for age, total energy intake, alcohol intake, and physical fitness. Over the 7-year follow-up, all smoking status groups gained weight, including continuous smokers and initiators. Weight gain was greatest among those who quit smoking. Weight gain attributable to smoking cessation was 4.2 kg for Whites and 6.6 kg for Blacks. Smoking had a small weight-attenuating effect on Blacks. No such effects, however, were observed among Whites. These results suggest, at least in younger smokers, that smoking has minimal impact on body weight. PMID- 9874913 TI - Personality predictors of mood related to dieting. AB - The clinical utility of a model of normal emotional functioning (vs. psychopathology) and the moderating effects of neuroticism (N) and extraversion (E) on mood were examined during a 6-week weight-loss trial. Participants were 40 obese women who completed measures of negative affect (NA) and positive affect (PA) weekly during the diet and measures of anxiety and depression (Beck Depression Inventory [BDI]) at pre-, mid-, and postdiet. Results indicated that (a) average NA and PA were each uniquely related to postdiet BDI scores, (b) N was significantly related to NA during the diet and postdiet BDI scores, and (c) N and E interacted to predict PA during the diet. The results suggest that assessment of personality and normal mood variation may be useful additions to weight-loss intervention and research. PMID- 9874911 TI - Cognitive processing, discovery of meaning, CD4 decline, and AIDS-related mortality among bereaved HIV-seropositive men. AB - This study investigated whether finding meaning in response to an HIV-related stressor was associated with changes in immune status and health. Forty HIV seropositive men who had recently experienced an AIDS-related bereavement completed interviews assessing cognitive processing and finding meaning after the loss and provided blood samples for a 2- to 3-year follow-up. AIDS-related mortality over an extended follow-up was determined from death certificates. As predicted, men who engaged in cognitive processing were more likely to find meaning from the loss. Furthermore, men who found meaning showed less rapid declines in CD4 T cell levels and lower rates of AIDS-related mortality (all ps < .05), independent of health status at baseline, health behaviors, and other potential confounds. These results suggest that positive responses to stressful events, specifically the discovery of meaning, may be linked to positive immunologic and health outcomes. PMID- 9874914 TI - Relationships between stages of change in cigarette smokers and healthy lifestyle behaviors in a population of young military personnel during forced smoking abstinence. AB - This study evaluated the relationships between stages of change and related health behaviors in a population of smokers forced to quit smoking. Participants were 10,136 Air Force recruits who were in basic military training (BMT) and who were not allowed to smoke because of a ban on smoking during BMT. Participants were surveyed about their smoking history, their motivation and readiness to remain smoke free after BMT, and their behavior on 5 target health areas. Results indicated that smoking history best predicted stage of change. Though there were few gender differences, several ethnic differences emerged. These results suggest that, although stages of change involve both a cognitive and behavioral component, removing the behavioral component (smoking) through a smoking ban does not reduce the construct validity of the transtheoretical model. PMID- 9874915 TI - Interpersonal ambivalence, perceived relationship adjustment, and conjugal loss. AB - Ambivalence is widely assumed to prolong grief. To examine this hypothesis, the authors developed a measure of ambivalence based on an algorithmic combination of separate positive and negative evaluations of one's spouse. Preliminary construct validity was evidenced in relation to emotional difficulties and to facial expressions of emotion. Bereaved participants, relative to a nonbereaved comparison sample, recollected their relationships as better adjusted but were more ambivalent. Ambivalence about spouses was generally associated with increased distress and poorer perceived health but did not predict long-term grief outcome once initial outcome was controlled. In contrast, initial grief and distress predicted increased ambivalence and decreased Dyadic Adjustment Scale scores at 14 months postloss, regardless of initial scores on these measures. Limitations and implications of the findings are discussed. PMID- 9874916 TI - Mild closed-head injury in children and adolescents: behavior problems and academic outcomes. AB - The issue of whether mild head injuries (HIs) in children cause behavior problems and poor scholastic performance is controversial. This study included 119 children (range = 8-16 years old) with HI, 114 with other injuries, and 106 with no injury (NI). Behavioral functioning was assessed with the Child Behavior Checklist; academic functioning, with school grades and standardized testing. Higher T scores were found for both injury groups versus NI participants on preinjury behavioral status. All 3 groups' behavioral scores decreased relative to baseline at 1 year. HI and NI groups did not differ in school grades or achievement testing either pre- or postinjury. These results are consistent with the conclusion that head injury of the mildest type does not increase the probability of new overt behavioral or academic problems. PMID- 9874917 TI - Data monitoring: a hypothesis-testing approach for treatment-outcome research. AB - Traditional inferential statistics require that hypotheses be evaluated at only 1 sample size. That is, researchers must choose how many participants will be included in a study before conducting analyses; they are not allowed to add data if initial results are not significant. This requirement forces researchers to choose among including more participants than necessary, risking inconclusive results, or violating the requirement by adding participants. This study presents a more flexible approach, called data monitoring, that allows repeating an analysis as the sample increases. First, the cost of the uncorrected data monitoring that researchers sometimes do is estimated. Second, the correction that is needed to allow data monitoring while holding an overall alpha at a desired level is estimated. Third, the power of data monitoring is compared with traditional approaches. This study also provides an example of the use of data monitoring. At least in some circumstances, data monitoring can reduce Type II error or the number of participants needed without sacrificing Type I error. PMID- 9874918 TI - Is there a role for continuation phase cognitive therapy for depressed outpatients? AB - Two pilot studies evaluated the rate of relapse or recurrence (i.e., major depressive disorder) after cognitive therapy (CT). Two sequential cohorts included outpatients who responded to acute phase CT (A-CT) and who agreed to monthly, treatment-free follow-up. In Study 1, the Kaplan-Meier technique estimated relapse and recurrence rates of 40% at 6 months, 45% at 8 months, 50% at 12 months, 67% at 18 months, and 74% at 24 months. In Study 2, responders to A CT received 8 months (10 sessions) of continuation phase CT (C-CT). In Study 2, relapse or recurrence was 20% at 6 and 8 months, 27% at 12 months, and 36% at 18 and 24 months after A-CT. An exploratory log-rank test showed that relapse or recurrence-free survival was greater in Study 2 than in Study 1. If replicated, this result suggests that C-CT can reduce depressive relapse or recurrence. Alternative explanations are presented. PMID- 9874919 TI - The crisis of elder care. PMID- 9874920 TI - Acceptance of nurse practitioners and physician assistants in meeting the perceived needs of rural communities. AB - Nurse practitioners and physician assistants have provided a partial solution to the shortage of primary care services in medically underserved rural areas. This paper describes the results of a study exploring community acceptance of nurse practitioners and physician assistants in rural medically underserved areas. Community acceptance in the context of this study implies not only satisfaction with care received, but also willingness of the community to support NP/PA practice through its infrastructure and encourage members to initially seek and continue to receive care from an NP or PA. Five focus groups were conducted in each of five rural medically underserved communities. The two most pervasive findings were the lack of previous exposure to NPs and PAs and the general belief that NPs and PAs would be accepted in these communities if certain conditions could be met. The theme of conditional acceptance included both personal and system factors. Personal factors included friendliness, competence, willingness to enter into the life of the community, and the ability to keep information confidential. System factors considered critical for acceptance included service type, integration with the existing health care system, cost, geographic proximity, and availability. The results of this study offer insight into community attitudes and suggest marketing strategies for those who plan to introduce NP or PA services into rural communities. PMID- 9874921 TI - The community health nursing implications of the self-reported health status of a local homeless population. AB - This study explored the personal characteristics and the health and health related concerns reported by members of the local homeless population in order to design population-specific health programming. The study also examined whether there were significant differences between homeless who are shelter residents and those who are not. An exploratory descriptive design was used to analyze retrospective data collected by a local County Health Department in interviews of 132 homeless adults. The demographic characteristics found reflect many common patterns: marked over-representation of males, mean age in the mid-thirties, education levels comparable to similar socio-economic groups, high unemployment rates, and low health insurance rates. One third of the sample reported self assessed health statuses of fair or poor. The most frequently identified physical health issue was joint problems, followed by cardiovascular disease. Depression was mentioned most frequently as a self-identified mental health problem. Loneliness was the number one fear identified. Chi Square analysis showed that homeless who did not stay in shelters were significantly longer term residents (p < 0.0001) of the community and reported fear of loneliness significantly more frequently (p < 0.01). This study identifies health concerns that local homeless people themselves find important and provides direction for development of sound population-specific health programming. PMID- 9874922 TI - Development of a community-based diabetes and hypertension preventive program. AB - The purpose of this project was to develop rapport with a Chinese Community Association and then establish preventive diabetic and hypertension programs with the Chinese in Chinatown, Hawaii. Subjects were recruited from this Chinese Community Association. Two hundred Chinese responded to the invitation. Among these, 75 individuals had either Type 2 diabetes, hypertension or both. Thirty six males and 39 females ranging in age from 51 years old to 96 years old (Mean = 71.76, SD = 9.58) participated. Surveys and educational programs were carried out in Chinese. Results were described in terms of quantitative measures (family support and health outcomes) and qualitative experiences (case studies). Eighty percent of participants had decreased their diastolic blood pressure from above 95 mmHg to below 90 mmHg and systolic blood pressure from above 155 mmHg to below 140 mmHg. Ninety-five (n = 71) percent of participants had maintained their glucose level within the 90 mg/dL to 150 mg/dL range with a mean reduction of 57.86 mg/dL in one year. The hardest thing for families was the glucose self monitoring. Case studies suggested that open-minded active listening and persistence formed the basis for developing a culturally sensitive community based self management program for chronic diseases. Collaboration among the community, public health nurses, and diabetes nurse educators facilitated the process of community education and health promotion. PMID- 9874923 TI - Acanthosis Nigricans as an early clinical proxy marker of increased risk of type II diabetes. AB - Diabetes affects 6% of the national population, yet approximately 50% of persons with diabetes remain undiagnosed and receive no treatment. In specific populations, Acanthosis Nigricans (AN) may serve as an early clinical proxy marker of increased risk of type II diabetes. The results of this pilot project to screen selected school age students in New Mexico for AN indicate that a large number of these students may be at increased risk for developing type II diabetes as young adults. The future burden of diabetes on individuals, families, communities, and health care systems may be greater than previously recognized. PMID- 9874924 TI - Abuse against women in rural Minnesota. AB - The purpose of this study was to examine the prevalence of physical, emotional, and sexual abuse in women seeking care in rural medical clinics and WIC supplemental food program sites, as well as to examine demographic and health characteristics related to abuse. This was a descriptive, cross-sectional study using a self-administered, anonymous survey. Data were collected in 8 medical clinics and 17 WIC supplemental food program sites in 9 counties of west-central Minnesota during January and February, 1997. The survey instrument asked brief screening questions on physical, emotional, and sexual abuse as well as demographic and health related questions designed to reflect the rural population and characteristics. The survey results showed that in the prior 12 months, 109 (6.6%) of the respondents reported physical abuse, 347 (21.1%) reported emotional/verbal abuse, and 35 (2.1%) reported sexual abuse. Proportional increases in risk of current abuse relative to married women were 2.1 for single, 2.5 for divorced, and 6.5 for separated women, after adjusting for other important variables. Single women with a recent status change had significantly higher risk of current abuse than women with a recent status change at all other marital status levels in adjusted analyses. Women who reported some form of abuse before age 18, and women reporting abuse as an adult prior to the last 12 months, were significantly more likely to report current abuse in all analyses. Prior abuse as an adult was the single best predictor of current abuse in a final logistic regression model. PMID- 9874925 TI - Parenting competence, social support, and self-esteem in teen mothers case managed by public health nurses. AB - The purpose of this descriptive study was to determine whether self-esteem, parenting competence, and social support for teenage mothers changed over the first 18 months of parenting when case managed by a public health nurse (PHN). A sample of 56 first-time teen mothers from a health department parenting project agreed to participate in the study. PHN case managers collected data close to the birth of the infants and at 6, 12, and 18 months. Demographic findings of teen mothers showed that the majority were below expected grade level, over half lived with parents, and over half were children of teen mothers. Outcome findings related to the infants revealed no delays in development as measured on the Denver Development Screening Test, adequate follow-up for identified health problems, and a high percentage of the children with up-to-date with immunizations. The research question findings showed a statistically significant drop in self esteem for the teens between birth and 6 months, and in social support between 6 and 18 months. No other findings were significant, but some trends appeared when the sample was divided by ethnicity, suggesting a need for closer follow-up for certain groups. Implications for public health nursing and nursing education are included. PMID- 9874926 TI - The relationship between confidence and accuracy in clinical assessment of psychiatric patients' potential for violence. AB - The authors studied the relationship between confidence and accuracy in clinical assessments of psychiatric patients' short-term risk of violence. At the time of entry to the hospital, physicians (N = 78) estimated the probability that each of 317 patients would physically attack other people during the first week of psychiatric hospitalization. The clinicians also indicated the degree of confidence they had in their estimates of violence potential. Nurses rated the occurrence of inpatient physical assaults with the Overt Aggression Scale. The results showed that when clinicians had a high degree of confidence, their evaluations of risk of violence were strongly associated with whether or not patients became violent. At moderate levels of confidence, clinicians' risk estimates had a lower, but still substantial relationship with the later occurrence of violence. However, when clinicians had low confidence, their assessments of potential for violence had little relationship to whether or not the patients became violent. The findings suggest that the level of confidence that clinicians have in their evaluations is an important moderator of the predictive validity of their assessments of patients' potential for violence. PMID- 9874927 TI - Reducing the hindsight bias utilizing attorney closing arguments. AB - In the legal system, jurors are asked to render a decision after the event in question has already occurred and the final outcome, typically negative, is known. This "after-the-fact" structure of the legal system makes jurors susceptible to a human judgment phenomenon known as hindsight bias. This study focused on reducing hindsight bias in a courtroom context by incorporating a debiasing strategy within the defense's closing argument. Subjects viewed one of three videotaped versions of plaintiff and defense closing arguments in a commercial litigation case (i.e., foresight condition, hindsight condition, and hindsight debiasing condition). Results indicate that the hindsight debiasing strategy was effective in reducing subject-juror hindsight bias. PMID- 9874928 TI - When believing is seeing: the effect of scripts on eyewitness memory. AB - Two studies examined the conditions under which event schema or scripts produce gap-filling errors in eyewitness accounts of a robbery. In Study 1, scripts for the robbery of a convenience store were identified. Results revealed high agreement among the 120 participants concerning the sequence of actions for such a robbery. Based on the information obtained in Study 1, participants in Study 2 (N = 144) viewed one of two sequences of slides depicting a robbery of a convenience store by a lone robber. In one sequence, three central script actions were omitted and in the other, three peripheral script actions were omitted. In addition, rate of exposure was varied (2 vs. 8 sec) as was the length of the retention interval (5 min vs. 1 week). As predicted, there was a higher rate of false recognition for central as opposed to peripheral actions, and this tendency was exaggerated for the longer retention interval. The theoretical and practical implications of the findings are discussed. PMID- 9874929 TI - The effects of juror anonymity on jury verdicts. AB - This study examined the effects of anonymity on jurors' verdicts and on jurors' feelings of accountability for their jury's verdicts. Twenty four-person anonymous juries and 20 four-person nonanonymous juries rendered individual and group verdicts for three student defendants charged with selling drugs on a school campus. When unanimous guilty verdicts were reached, juries imposed one of five punishments. Finally, jurors completed postdeliberation opinion and accountability questionnaires. As predicted, anonymous juries showed a higher rate of conviction (70%) than did nonanonymous juries (40%) when the evidence against the defendant was strong, supporting the hypothesis that anonymity would have a greater effect for situations in which there was relatively strong evidence of the defendant's guilt. Anonymous juries imposed the harshest punishment (expulsion) significantly more often than did nonanonymous juries. Contrary to predictions from differential self-awareness theory, anonymous juries did not report feeling less accountable than did nonanonymous juries. However, anonymous juries did see the process as significantly more fair than did identifiable juries. PMID- 9874930 TI - Issues in clinical integration. PMID- 9874931 TI - Integrating a clinical service line for maternal-child health. AB - Clinical integration is a developmental process that calls on the combined resources, expertise, and knowledge of individuals who may be working together for the first time. The process is dependent on the acknowledgment of the value of integration and a shared vision for the service line. By using a strategic thinking and planning process, the service line can be reshaped to function as an integrated system of programs and services that is sensitive to needs at a local level. While the task of integration is not an easy one, the outcomes of the process lead to a synergy that cannot be realized in any other manner. PMID- 9874932 TI - Case management as a tool for clinical integration. AB - Case management has evolved to meet the challenges of contemporary health care. As an effective strategy to promote clinical integration within health care systems, case management brings together population and individual approaches to development and coordination of evidence-based, outcome-focused health, disease, and illness management programs and services. A three-level prevention-focused model for health care management across the continuum of care is presented. PMID- 9874933 TI - Clinical integration across multiple hospitals: the agony, the ecstasy. AB - The author shares one system's experience with the clinical integration of multiple facilities over several years. This inside view relates the good, the bad, and the ugly; the difficulties and the successes; and the agonies and the ecstasies. The author arrives at the conclusion that all the pain and anguish are worth the rewards of an integrated system that is well positioned in the marketplace to meet the mission and vision of a community health care system committed to thriving in the 21st century. PMID- 9874934 TI - Developing an integrated model for outcomes management. AB - In the complexity of integrated delivery systems, the triad of clinicians, financial analysts, and information management personnel can partner to form an effective force in driving outcomes management efforts. Health care systems can take the opportunity to rethink outcomes management strategies with an emphasis on monitoring cost and quality simultaneously. Advanced practice nurses, with a wide breadth of knowledge obtained through higher education, are positioned to be active, effective team members at all points in the outcomes management process. PMID- 9874935 TI - The development of care management systems to achieve clinical integration. AB - With development of managed care markets, health care delivery systems face increasing clinical and financial risk. For an integrated delivery system to survive, strategies for clinical integration and care management are essential. CareMap tools, collaborative practice groups, and case management serve as the foundation to accomplish care management over the health continuum. Coordination of care, within an institution and across traditional health settings, to achieve the best clinical and cost outcomes is the goal. The article discusses strategies for clinical integration, categories for measurement of performance, and the need to incorporate automated solutions into the strategic business plan. PMID- 9874936 TI - Clinical integration in a population-based planning framework. AB - Population-based planning provides context and scope to the goal of clinical integration. It also is a transforming process for organizations and programs devoted to a hospital-centric care system, as well as source of language that can support collaboration among buyers, payers, providers, and community groups. At the clinical care level, population-based planning recognizes the differences in how people perceive health and use health care services, as an important corollary to understanding specific diseases and clinical service needs. PMID- 9874937 TI - Ethical balance versus an ethical anomaly. AB - "A Practice-Based Bioethic" is a regular column in Advanced Practice Nursing Quarterly. A practice-based approach is derived from, and therefore is intended to be appropriate to, the situation of a patient, the purpose of the health care setting, and the role of the nurse. It is based on a shared state of awareness, the foundation on which ethical interactions between nurse and patient occur. PMID- 9874938 TI - Breast cancer: agenda setting through activism. AB - Breast cancer has long been one of the leading causes of death among women in the United States. The disease did not gain serious attention in the public policy arena, however, until the 1990s. Using Kingdon's agenda-setting model as a framework, this article describes how breast cancer moved to a place of prominence on the national health care agenda. The role of breast cancer activists in this effort is examined. Suggestions are then made concerning why and how advanced practice nurses might effectively influence the health policy agenda through political activism. PMID- 9874939 TI - Opening to the sacred: intentional use of music to engage the spiritual dimension. AB - Music is a patterned temporal art characterized by its pulse, rhythm, and melody. It can be intentionally used as a vehicle or gateway to the nonrational as contrasted to an irrational dimension of human experience. This realm of experience frequently provides access to the sacred. The indissoluble unity of body-mind-spirit has been a tenet of creative artists for centuries, and is now gaining the attention of health care professionals. This article explores the conceptual and applied potential of integrating music as a spiritual practice in skilled and compassionate health and illness care. PMID- 9874940 TI - Caring theory: a framework for advanced practice nursing. AB - This article provides a brief review of Watson's theory of caring and demonstrates its usefulness as a framework for advanced practice nursing. Two case studies, a recently diagnosed patient who is positive for the human immunodeficiency virus and a patient terminally ill with acquired immunodeficiency syndrome, are reviewed using the carative factors as a model for providing holistic, patient-focused nursing care. The value of this exercise lies in the demonstration of the applicability of caring theory as a framework for both symptom management and palliative nursing care. PMID- 9874941 TI - Clinical integration: partnerships for care. AB - Integration means establishing new relationships and building partnerships with other disciplines and with patients. Nurses must be about this work if they are to ensure a role in the future. PMID- 9874942 TI - Nursing by agreement: a contractarian perspective on nursing ethics. AB - Contractarianism has primarily been a political theory. A good argument can be made for its application to individual and professional ethics. A contractarian perspective on nursing ethics not only provides useful insights into the central relationship in nursing--the nurse and the patient--but also allows understanding of that relationship in the context of other relationships involving the nurse, the nursing profession, and the health care organization. While the nurse-patient contract is limited, its greater moral depth gives it moral priority. PMID- 9874943 TI - Investing or discounting self: are moral decisions shaped by conditions in the workplace? AB - Modern socioeconomic trends contribute to the complexity and diversity of ethical issues encountered in clinical nursing practice. Despite growing interest in ethical decision making among nurses, little is known about the contextual influences. The article describes an exploratory study of how nurses make ethical decisions and what factors hindered or helped them to act on their moral choices. Two central constructs were identified: investing and discounting self. The article explains essential subprocesses of these constructs within the context of clinical nursing practice. PMID- 9874944 TI - Revisiting the call to care: an ethical perspective. AB - As the 21st century rapidly approaches, a reexamination of nursing's call to care is timely. Caring, long claimed as central to nursing practice, seems to be in jeopardy. The article presents evidence that nurses do care for their patients. Moreover, caring is essential to the moral integrity of their practice. The article calls on nurses to put patients before convenience and to remember that without patients there would be no profession. PMID- 9874945 TI - Genetic testing for cancer: issues and ethical implications. AB - The explosive progress that has been made in cancer genetics in the past few years has given rise to new hope. These promising discoveries have also given rise to considerable uncertainty and debate concerning how this knowledge should be applied and who should have ownership of its products and effects. The medical, psychological, social, legal, and ethical implications of genetic testing for cancer, specifically breast and ovarian cancers, are discussed. The ethical principles of confidentiality, justice, beneficence, and autonomy as they relate to cancer predisposition testing also are explored. Recommendations are made for nursing practice, education, and research. PMID- 9874946 TI - When pregnant women and their physicians disagree on the need for cesarean section: no simple solution. AB - This article provides an analysis of why women might refuse cesarean sections that are advised by their physicians. It presents a review of court cases, ethical opinions, and recommendations in the literature and lists the legal requirements for informed consent. The author delineates the process of weighing whether or not a court order should be requested. PMID- 9874947 TI - Managed care and ethical implications in telephone-based health services. AB - Telephone-based health services is a fast growing subspecialty of nursing practice. It is used in demand management and managed care organizations in a highly unregulated environment. The article presents background information and identifies the need for systematic studies of ethical implications inherent in telephone-based practice. Conclusions suggest that nurses need to become actively involved in ethical decision making related to this area of practice. PMID- 9874948 TI - With the ethical agreement: where are you now? AB - "A Practice-Based Bioethic" is a regular column in Advanced Practice Nursing Quarterly. A practice-based approach is derived from, and therefore is intended to be appropriate to, the situation of a patient, the purpose of the health care setting, and the role of the nurse. It is based on a shared state of awareness on which ethical interactions between nurse and patient occur. PMID- 9874949 TI - Death, denial, and defeat: older patients and advance directives. AB - This article describes a phenomenological study involving 17 older persons who shared their reasons for not having formalized their end-of-life decisions. The study found they vacillated between fear and acceptance of death, trust and mistrust that others would or would not act according to their wishes, the assumption of immortality or the inevitability of death, and the denial or lack of knowledge of the necessity for legalizing their wishes. A discussion of the implications of these findings for nurse practitioners is presented. PMID- 9874950 TI - The impact of reimbursement policies on the viability of certified nurse midwifery practices. AB - The article reports on a study of the effects of reimbursement policies on the ability of certified nurse midwives (CNMs) to maintain a viable practice. In particular the impact of reimbursement policies on the ability to maintain and seek a client base and the overall financial viability of the practice was explored. Neither type of reimbursement statute (private or Medicaid) had an effect on the ability of CNM practices to maintain an appropriate number of clients. Moreover, neither reimbursement statute had a relationship to how a CNM is paid, the type of benefits received, and whether or not a practice covered its expenses. PMID- 9874951 TI - Children with severe and profound disabilities and the issue of social justice. AB - Children with severe and profound disabilities are often seen as less worthy of health care resources. In times of managed care and utilitarian allocation decisions, the return on investment of resources is not easily appreciated for children with severe disabilities. This article discusses the quality-of-life considerations, societal attitudes toward people with disabilities, and the tension of the technological imperative. Advanced practice nurses are in a position to advocate for the needs of children with severe disabilities. There is a chance that an ugly history could be repeated if the lives of children with disabilities are not valued. PMID- 9874952 TI - Examination of ethical practice in nursing continuing education using the Husted model. AB - Beliefs about human nature, adult education, adult learners, and moral commitment are at the heart of the educator-learner agreement. In continuing nursing education, it is the point where professional values, morals, and ethical principles meet. Using Husteds' bioethical decision-making model, the values, beliefs, and actions within the educator-learning agreement are identified and organized by the bioethical standards. By relating the bioethical standards to practice, continuing nurse educators can find their own basis for practice and work toward attaining a consistent professional ethical orientation. PMID- 9874953 TI - Physician-assisted suicide and older patients' perceived duty to die. AB - Physician-assisted suicide (PAS) may soon be legal in the United States. The legal option of PAS will place certain hardships on older persons. Many older individuals may feel a duty to die to avoid becoming a burden to their families. Moreover, the suicide option becomes attractive because medical care at the end of life is often fragmented, impersonal, and unable to meet the needs of the dying individual. To make suicide less attractive, nurses must help restructure the care of the dying to help meet their needs and reduce the burdens of caregivers. PMID- 9874954 TI - Moral evaluation and concepts of health and health promotion. AB - This article considers the moral implications of some of the ways in which society, including nursing, thinks about health and health promotion. How we think about health shapes what we describe as healthy or unhealthy, as well as those factors that promote or hinder the attainment and maintenance of health. In turn, these concepts lead to actions that have moral significance. These issues are of particular import to nursing because health is central to what we understand our aims to be. To this end, the article addresses definitions of health, the relationship between morality and health, health promotion and the moral harms following from some of those practices, and the challenges to nursing. PMID- 9874955 TI - Clinical nurse specialist prescriptive authority and the legislative process. AB - Despite certification and graduate education, clinical nurse specialists are denied prescriptive authority in many states. Restrictions on their scope of practice and prescriptive authority are barriers to potential contributions to the health care delivery system. The article describes the experience of the New Mexico Council of Clinical Nurse Specialists in the process of obtaining prescriptive authority. It discusses council formation, lobbying activities, the legislative process, and ways to become politically involved. Clinical nurse specialists are encouraged to become politically active and work through their state nurses association to change restrictive policies. PMID- 9874956 TI - Ethics in health care: are nurses at the table? AB - The ethical issues emerging in a capitated health care marketplace will demand energy, commitment, and assertive response on the part of nurses in every arena of practice. PMID- 9874957 TI - An international perspective: APN credentialing. AB - This article identifies the global environment for the development and regulation of advanced practice nursing; provides a conceptual framework within which to examine regulatory concepts and methods and identify contemporary issues; introduces the worldwide diversity of nursing practice roles, standards, and regulatory systems, including some examples; and proposes a role for American nurse experts in the development of an international movement in advanced practice and in credentialing. PMID- 9874958 TI - Striving for quality in advanced practice nursing education. AB - The issue of establishing and maintaining high-quality advanced practice educational programs, practice competencies, and certification criteria is currently the focus of much debate and discussion within nursing. This article focuses on the issue of accreditation for advanced practice nursing educational programs in four areas: (1) history of accreditation in nursing education; (2) accreditation of educational programs for clinical nurse specialists, nurse midwives, and nurse anesthetists; (3) accreditation of programs for nurse practitioners; and (4) current and future educational issues and challenges. To participate in shaping the changes occurring in the current health delivery system, the nursing profession and advanced practice nursing educators must present a strong, visionary, and collaborative voice concerning quality educational programs. PMID- 9874959 TI - Politicization of health professions regulation. AB - Professions evolve through five stages of development--ideology, implementation, evaluation, incorporation, and maturation. Further, the development and implementation of health professions regulation is fraught with political overtones. This article provides an analysis of regulation in advanced practice nursing as it relates to the other health professions and presents efforts to reform health professions regulation using strategies offered by major philanthropic foundations. PMID- 9874960 TI - Still spending dollars, still searching for sense: advanced practice nursing in an era of regulatory and economic turmoil. AB - Regulatory and market forces are dramatically affecting the practice prospects for advanced practice nurses (APNs). Examples include the designation of APNs as primary care providers by for-profit capitated systems, the elimination of "geographic" practice boundaries by the advent of telepractice, and the revision of governmental reimbursement provisions for entire categories of APNs. Educational, political, and economic challenges necessitate an increased APN leadership role in national and state policy reform efforts. PMID- 9874961 TI - Advanced practice nursing certification: where do we go from here? AB - Certification requirements for advanced practice nurses (APNs) vary between APN specialties and the certification agencies. This variability perpetuates confusion for APNs, the health care system, and the public. Recommendations are made for the certification of APNs: (1) standardization of the certification process for APNs, (2) consistency in the requirement for graduate level education for all APNs, (3) regulation requiring certification for practice in all states, and (4) nurse practitioner (NP) program accreditation or preaccreditation for eligibility to take the certification examination. To support these recommendations, two questions are considered: Where are we now in certification? Where do we need to go? PMID- 9874962 TI - Using research to influence the regulatory process. AB - The need for sound research data to support regulatory decisions related to advanced nursing practice has never been greater. Although advanced practice nurses (APNs) are frequently exhorted to engage in research and to become involved in the political arena, there is little published evidence that nursing research has been used to influence health policy. This article presents two case examples of the effective use of research findings to influence regulatory policy in two different states. Factors involved in using research to shape policy are discussed, including recommendations for nursing policymakers and researchers. Research is a powerful tool that APNs can and should use to make a difference in regulatory legislation. PMID- 9874963 TI - The nurse as cynic--etiology and Rx. AB - "A Practice-Based Bioethic" is a regular column in Advanced Practice Nursing Quarterly. A practice-based approach is derived from, and therefore is intended to be appropriate to, the situation of a patient, the purpose of the health care setting, and the role of the nurse. It is based on a shared state of awareness on the basis of which ethical interactions between nurse and patient occur. PMID- 9874964 TI - Credentialing of certified registered nurse anesthetists. AB - Certified registered nurse anesthetists (CRNAs) have provided anesthesia services in this country since the mid-1800s. As the earliest advanced practice nurse specialists, CRNAs have been leaders in the development of educational programs, accreditation, certification, and recertification. This article will describe the history of credentialing in nurse anesthesia and the achievements in the development of a valid and highly respected credentialing mechanism. Recertification and hospital credentialing are also discussed. PMID- 9874965 TI - Core competency-based education, certification, and practice: the nurse-midwifery model. AB - It has been more than 70 years since the first nurse-midwives came to practice in the United States, and over 20 years since the American College of Nurse-Midwives (ACNM) established nationally recognized standards and core competencies for nurse-midwifery education, certification, and practice. Certified nurse-midwives are licensed in all 50 states and in the District of Columbia (DC) and have prescribing privileges in 45 states plus DC. More recent standard-setting activities include the establishment of a national mechanism for continuing competency assessment; adoption of the bachelor's degree as the minimum academic requirement for certification; development of national standards for education and certification of direct entry midwives; and placement of an 8-year time limit on certificates. PMID- 9874966 TI - Nurse practitioner certification: benefits and drawbacks. AB - This article discusses the history of certification process and distinguishes and analyzes past and present uses of the certification process. It explores changes in the regulatory process and the impact on nurse licensure and evaluates past, present, and future uses of certification related to advanced practice, credentialing, and the scope of nursing practice. PMID- 9874967 TI - Advanced practice nursing in Texas: the interface of accreditation, regulation, and certification. AB - The rapidity and degree of development of regulation by boards of nursing of advanced practice nurses (APNs) and programs for their preparation has varied from state to state over the past 20 years. In general, boards have authority only to regulate advanced practice through the recognition of APNs and the setting of standards and scope of their practice. In the early 1990s, lack of consistent APN educational program standards and experiences and criteria for recognition of APNs was problematic at the levels of accreditation, certification, and regulation. The interdependency of these concepts as well as the necessity and difficulty of linking them in a consistent, effective manner has become increasingly evident. The Texas Board of Nurse Examiners has been involved in deriving a model designed to ensure the education and recognition of APNs with emphasis on both professionalism and public safety. Because Texas is unique in size and geography, it mirrors many problems and issues of both large and small states. PMID- 9874968 TI - Mutual recognition: response to the regulatory implications of a changing health care environment. AB - Mutual recognition offers a regulatory solution for dealing more effectively with the changes that have occurred, and continue to occur, in health care. To meet the demands of the integrated delivery system, the challenges of technological advances, and the public need for access to nursing care, nursing regulators have explored ways to make state boundaries more permeable while maintaining critical standards for consumer protection. This article discusses the evolution of regulation, the dilemma caused by the changing practice environment, why mutual recognition was chosen as the model for regulatory reform, and how mutual recognition will work from the perspective of the National Council of State Boards of Nursing. PMID- 9874969 TI - A new age for regulation. AB - Regulation will require a new approach for health professions. No longer can they approach the process with notions of exclusivity, exclusion, or independence from each other. PMID- 9874970 TI - Chief nurses from the EU countries met together. PMID- 9874971 TI - Fairness at work. PMID- 9874972 TI - Making shared governance work. PMID- 9874973 TI - Can we really blow the whistle? PMID- 9874974 TI - Implementing clinical effectiveness. PMID- 9874975 TI - Never mind the quality: what about the outcome? PMID- 9874976 TI - Structuring health care for the future. PMID- 9874977 TI - A touch of care. PMID- 9874978 TI - Evidence-based practice. PMID- 9874979 TI - Primary care groups. PMID- 9874980 TI - Breaking the code of silence. PMID- 9874981 TI - Have hospices lost their way? PMID- 9874982 TI - Engaging support. PMID- 9874983 TI - Patient ratings. PMID- 9874984 TI - Nursing: A powerful European force. PMID- 9874985 TI - Who cares about nursing. Interview by Tom Keighley. PMID- 9874986 TI - Global links. PMID- 9874987 TI - Continuing to care. PMID- 9874988 TI - Keeping in good spirits. PMID- 9874989 TI - Backs to the future. PMID- 9874990 TI - Children's services: why managers lack power. PMID- 9874991 TI - Crossing professional boundaries. PMID- 9874992 TI - Satisfaction and motivation. PMID- 9874993 TI - The American dream or British vision. PMID- 9874994 TI - Nursing in retreat. Interview by Tom Keighley. PMID- 9874995 TI - Political leadership in nursing. PMID- 9874996 TI - Ethical issues in health care rationing. PMID- 9874997 TI - Nursing's future the challenge. PMID- 9874998 TI - Healing the ailing heart: the power of spiritual care. PMID- 9874999 TI - The 'Y2K bug': how to ease hospital equipment into the next millennium. AB - The advent of the 21st century is unlikely to wreak as much technologic havoc as the doomsayers have predicted. The Y2K problem is addressed by new programs that use a four-digit year code. Although hospitals must prioritize technology for Y2K compliance, they also need to address the issue actively: 2000 is less than 18 months away! PMID- 9875000 TI - The practice of nursing theory in the operating room. AB - Nursing theory is a set of ideas, experiences, or observations regarding how and why nurses engage in certain activities and exhibit various behavioral attributes in providing patient care. The individualized caregiving activities perioperative nurses express during all phases of the surgical experience are supported by nursing theory. PMID- 9875001 TI - Developing a pain management protocol in the PACU. PMID- 9875002 TI - Preventing wrong-site surgery. PMID- 9875003 TI - Avoiding surgical site infections. PMID- 9875004 TI - Laparoscopic fundoplication. PMID- 9875005 TI - Privacy and confidentiality: what it is--what it is not (Part 1). PMID- 9875006 TI - Biological warfare: what happens if we are attacked? PMID- 9875007 TI - Operating room fires: how to prevent and minimize spread. AB - To prevent fires, a simple rule to remember is the "fire triangle" of oxygen, heat, and fuel. Minimizing oxygen enrichment will reduce the fire hazard. Proper protocols need to be established for the handling of heat-bearing units, such as electrosurgical pencils, electrocautery units, and lasers. PMID- 9875008 TI - Are "barrier" drapes cost effective? AB - When assessing the effectiveness of "barrier" materials, it is important to analyze other variables that can influence the rate of infection. The real issue at hand is not one of reusable versus disposable, but rather the cost effectiveness and expense associated with the use of barrier versus nonbarrier drapes. PMID- 9875009 TI - Medication errors--a system problem. AB - With medication errors, a more productive approach is to look at what, rather than who, caused the error. Unclear orders, both written and verbal, need to be clarified: Never make assumptions about the drug, dose, route, or frequency. Nurses must be assertive and join forces with pharmacists, physicians, risk managers, and quality improvement professionals to make a difference in error prevention and medication safety. PMID- 9875011 TI - Finding ways to encourage health care workers to wash hands often. Product placement and health care worker education on compliance were not enough to change behaviors. PMID- 9875010 TI - When your client in the surgical suite is Mexican American. AB - Although many Mexican Americans have assimilated into American culture, others have retained their cultural health beliefs about illness and medical treatment. Incorporating family members into the plan of care will greatly enhance compliance and health behaviors. The potential good of folk medicine should not be discounted; try to incorporate it into the existing plan of care. PMID- 9875012 TI - Privacy and confidentiality: when and how you must disclose (Part 2). PMID- 9875013 TI - Pretrial liaison between doctors in alleged child abuse. PMID- 9875014 TI - Attention deficit disorder with developmental coordination disorders. AB - AIMS: To analyse the contribution of certain social, familial, prenatal, perinatal, and developmental background factors in the pathogenesis of deficits in attention, motor control, and perception (DAMP). METHODS: A population based case-control study was carried out with 113 children aged 6 years, 62 diagnosed with DAMP and 51 controls without DAMP. The children's health and medical records were studied and their history with regard to background factors was taken at an interview with the mother using a standardised schedule. Familial factors, possible non-optimal factors during pregnancy (including smoking), developmental factors (including early language development), and medical and psychosocial data were scored in accordance with the reduced optimality method. RESULTS: Low socioeconomic class was common in the group with DAMP. Familial language disorder and familial motor clumsiness were found at higher rates in the DAMP group. Neuropathogenic risk factors in utero were also more common in the children with DAMP. Maternal smoking during pregnancy appeared to be an important risk factor. Language problems were present in two thirds of the children with DAMP. Sleep problems and gastrointestinal disorders, but not atopy or otitis media, were significantly more common in the DAMP group. CONCLUSIONS: Prenatal familial and neuropathogenic risk factors contribute to the development of DAMP. Primary prevention, such as improved maternal health care and early detection or treatment, or both, of associated language problems appear to be essential. PMID- 9875015 TI - Parental and professional perception of need for emergency admission to hospital: prospective questionnaire based study. AB - AIM: To compare views of parents, consultants, and general practitioners on severity of acute illness and need for admission, and to explore views on alternative services. METHOD: Prospective questionnaire based study of 887 consecutive emergency paediatric admissions over two separate three week periods in summer and winter of five Yorkshire hospitals, combined with a further questionnaire on a subsample. OUTCOME MEASURES: Parental scores of need for admission and parent and consultant illness severity scores out of 10. Consultant judgment of need for admission. Alternatives to admission considered by consultants and, for a subsample, by parents and family GP. RESULTS: Ninety nine per cent of parents thought admission was needed. Parents scored need for admission more highly than severity of illness with no association observed between severity and presenting problem or diagnosis. High parental need score was associated with a fit, past illness, and length of stay. Consultant illness severity scores were skewed to the lower range. Consultants considered admission necessary in 71%, especially for children aged over 1 year, presentation with breathing difficulty or fit, and after a longer stay. More admissions in the evening were considered unnecessary as were admissions after longer preadmission illness, gastroenteritis, or upper respiratory tract infection. Of a subsample of parents, 81% preferred admission during the acute stage of illness even if home nursing had been available. Similar responses were obtained from GPs. Alternative services could have avoided admission for 19% of children, saving 15.6% of bed days used. CONCLUSIONS: Medical professionals and parents differ in their views about admission for acute illnesses. More information is needed on children not admitted. Alternative services should take account of patterns of illness and should be acceptable to parents and professionals; cost savings may be marginal. PMID- 9875016 TI - Medical and social factors associated with the admission and discharge of acutely ill children. AB - AIM: To examine medical and sociodemographic factors involved in acute paediatric admission. To compare outcome of admission with factors present at time of admission. METHODS: Prospective questionnaire based study of 887 consecutive emergency general paediatric admissions to five Yorkshire hospitals during two separate three week periods in summer and winter. MAIN OUTCOME MEASURES: Discharge diagnosis, length of stay. RESULTS: Most admissions (53%) occurred "out of hours" with a peak during the evening. Two thirds (64%) of patients were under 3 years of age and clinical problems varied with age. Self referral via an accident and emergency department occurred in one third and was more likely after a fit in older children and in more socioeconomically deprived children. The most frequent presenting problems were breathing difficulty (24%), fit (16%), and feverish illness (15%). One quarter (24%) were discharged within 24 hours and 61% spent, at most, one night in hospital. Length of stay was shorter for night admissions and longer for children with a discharge diagnosis of asthma. Although most children had mild, self limiting illnesses, serious illness was subsequently found in 13% and could not be predicted from the presenting problems. CONCLUSIONS: Current demand on emergency paediatric admission is mainly from young children with mild self limiting illnesses who spend one night or less in hospital. Changes in delivery of care to acutely ill children must take account of the pattern and nature of presenting problems and be rigorously audited to ensure that improvements in the health of children continue. PMID- 9875017 TI - Systematic review of the treatment of upper respiratory tract infection. AB - OBJECTIVES: To assess the risks and benefits of antibiotic treatment in children with symptoms of upper respiratory tract infection (URTI). DESIGN: Quantitative systematic review of randomised trials that compare antibiotic treatment with placebo. DATA SOURCES: Twelve trials retrieved from a systematic search (electronic databases, contact with authors, contact with drug manufacturers, reference lists); no restriction on language. MAIN OUTCOME MEASURES: The proportion of children in whom the clinical outcome was worse or unchanged; the proportion of children who suffered complications or progression of illness; the proportion of children who had side effects. RESULTS: 1699 children were randomised in six trials that contributed to the meta-analysis. Six trials were not used in the meta-analysis because of different outcomes or incomplete data. Clinical outcome was not improved by antibiotic treatment (relative risk 1.01, 95% confidence interval (CI) 0.90 to 1.13), neither was the proportion of children suffering from complications or progression of illness (relative risk 0.71, 95% CI 0.45 to 1.12). Complications from URTI in the five trials that reported this outcome was low (range 2-15%). Antibiotic treatment was not associated with an increase in side effects compared with placebo (relative risk 0.8, 95% CI 0.54 to 1.21). CONCLUSIONS: In view of the lack of efficacy and low complication rates, antibiotic treatment of children with URTI is not supported by current evidence from randomised trials. PMID- 9875018 TI - The divergent ventilatory and heart rate response to moderate hypercapnia in infants with apnoea of infancy. AB - BACKGROUND: Inspired CO2 is a potent ventilatory stimulant exhibiting a paradoxical inhibitory effect on breathing at high concentrations. Severe respiratory depression as a result of CO2 rebreathing during sleep has been implicated as a possible trigger factor in sudden infant death syndrome (SIDS). OBJECTIVE: To investigate the ventilatory and heart rate (HR) responses to inhaled CO2 in infants with apnoea of infancy, a group believed to be at increased risk of SIDS. STUDY DESIGN: Thirty one infants with severe sleep related apnoea, 31 infants with mild recurrent apnoea, and 31 age and sex matched controls for the infants with severe sleep related apnoea were studied. HR was computed from digitised RR intervals, "ventilation" was recorded by inductance plethysmography, and PCO2 and PO2 were monitored by transcutaneous electrodes. The ventilatory and HR responses to CO2 were expressed as percentage increase in ventilation and change in HR/unit change in transcutaneous PCO2. RESULTS: The mean increase in transcutaneous PCO2 during CO2 challenge (0.45 kPa = 3.4 mm Hg) resulted in a mean increase in ventilation of 291%/1 kPa (7.3 mm Hg) increase in transcutaneous PCO2, with no difference between the groups. A significant difference between infants with severe sleep related apnoea and mild recurrent apnoea versus controls (p < 0.02, p < 0.01, respectively) was found in their HR response to CO2 challenge: HR decreased in 12 severe sleep related apnoea infants and 10 infants with mild recurrent apnoea, but only in two controls. CONCLUSION: Infants with apnoea of infancy frequently show a paradoxical decrease in HR during CO2 challenge, possibly because of an insufficient ability to mobilise cardiovascular defence mechanisms when challenged with hypercapnia. PMID- 9875019 TI - Mutation analysis in 57 unrelated patients with MPS II (Hunter's disease). AB - Genomic DNA from 57 unrelated MPS II (Hunter's disease) patients was analysed for mutations of the iduronate sulphatase (IDS) gene. The aim of the study was threefold: to identify the primary genetic lesion in patients, to investigate the correlation between genotype and phenotype, and most importantly, to provide reliable carrier testing for female members once the family mutation was identified. In 42 patients, point mutations were identified involving single base substitutions, deletions, or insertions. These included four new nonsense mutations (R8X, C84X, E245X, Y466X), six new missense mutations (D45N, N115Y, P228L, P266R, E434K, I485K, W502C), three new insertions (c70C71ins, c652C654ins, c709G710ins), six new deletions (c500delC, c705delC, c1023delA, c1049delA, c1141delC, c1576delG), and five new mutations involving splice sites (IVS1-2 a- >g, IVS2-10 t-->g, IVS5 + 2 t-->g L236L, IVS7 + 2 t-->c). One patient had a new seven base deletion in exon 9 (c1482-1488del). Four patients were shown to have complete deletions of the IDS gene and two deletions involved one or more exons. Previously described mutations present in these patients were Q80X, P86L, R172X, G374G, S333L, R443X, and R468Q. In eight patients, no mutation was detected throughout the entire coding region. Most mutations that result in MPS II appear to be unique. Absence of the probands' mutations in eight of nine maternal grandmothers suggests many mutations have arisen recently. Prediction of the clinical phenotype from the identified genotype was difficult in some families, and further studies using reverse transcription polymerase chain reaction are needed to confirm the predicted effects on the IDS mRNA suggested by genomic analysis. PMID- 9875020 TI - Thyroid dysfunction in Down's syndrome: relation to age and thyroid autoimmunity. AB - BACKGROUND: The prevalence of thyroid disease is increased in Down's syndrome. Most available data come from cross sectional studies. AIMS: To study longitudinally thyroid function in patients with Down's syndrome in Uppsala county (85 patients) up to the age of 25 years. METHODS: Observational study based on yearly follow up in a children's clinic. Thyroid function tests were performed at each visit to the clinic. RESULTS: Hypothyroidism was found in 30 and hyperthyroidism was found in two of the 85 patients. No sex difference was seen. Half of the patients with hypothyroidism acquired the condition before the age of 8 years, but only one of them displayed thyroid autoantibodies at diagnosis. Most patients who developed hypothyroidism after this age had thyroid autoantibodies. In the prepubertal patients with hypothyroidism, growth velocity was lower during the year before the start of thyroxine treatment than during the year after treatment began; it was also lower than that of sex and age matched euthyroidic children with Down's syndrome. CONCLUSION: Thyroid dysfunction in patients with Down's syndrome is common in childhood. Consequently, annual screening is important. Autoimmune thyroid disease is uncommon in young children with Down's syndrome but is common after 8 years of age. PMID- 9875021 TI - Antineutrophil cytoplasmic antibodies in Wegener's granulomatosis. AB - The prevalence of antineutrophil cytoplasmic antibodies (ANCA) was studied in 12 children with Wegener's granulomatosis. The serum samples were taken in the active phase of disease and were screened for ANCA by indirect immunofluorescence with normal neutrophils and enzyme linked immunosorbent assay (ELISA) using crude neutrophil extract, proteinase 3, myeloperoxidase, cathepsin G, lactoferrin, and elastase as antigens. Of these 12 patients, 10 wre positive for ANCA in the active phase of their illness, and they showed a predominantly cytoplasmic ANCA staining pattern on indirect immunofluorescence. There were high titres of ANCA directed against crude neutrophil extract, proteinase 3, myeloperoxidase, and cathepsin G. IgM isotypes occurred as commonly as IgG isotypes. Therefore, screening for ANCA is usually but not invariably positive in children with Wegener's granulomatosis. Specific diagnosis still relies on clinical and pathological features, and the value of ANCA in the diagnosis of paediatric Wegener's granulomatosis requires further study. PMID- 9875022 TI - Suprasternal Doppler ultrasound for assessment of stroke distance. AB - An assessment of a non-invasive technique for measurement of stroke distance was made using a portable Doppler ultrasound machine. The aim was to determine the measurement error of repeated stroke distance measurements (Within-observer variability) and to assess measurement agreement between two operators (between observer variability). The measurement error (within-observer variability) for both operators was similar at approximately 2 cm. However, the measurements of the two operators (between-observer variability) did not agree well. Using the mean (SD) of three readings by each operator, the mean difference between the operators was -0.21 cm (1.96) giving a 95% confidence interval for the differences of -4.0 to +3.6 cm. There were significant positive and negative correlations between stroke distance and a variety of variables (age, height, weight, heart rate), but the relations were weak. The results indicate that the Doppler ultrasound technique for measurement of stroke distance would best be used to study trend changes in an individual patient, or subject, by a single operator. PMID- 9875023 TI - Incidence of acute respiratory distress syndrome: a comparison of two definitions. AB - OBJECTIVES: To determine the incidence and outcome of acute respiratory distress syndrome (ARDS) in children by comparing two commonly used definitions: the lung injury score and the American-European Consensus Conference definition. The causes and risk for developing ARDS were also studied. METHODS: Part prospective and retrospective analysis of 8100 consecutive hospital admissions from 1 June 1995 to 1 April 1997. RESULTS: Twenty one patients fulfilled the criteria for ARDS. Both definitions identified the same group of patients. The incidence was 2.8/1000 hospital admissions or 4.2% of paediatric intensive care unit admissions. The main causes were sepsis and pneumonia. Mortality was 13 of 21. Factors predicting death were a high admission paediatric risk of mortality (PRISM) score (30.38 v 18.75) and the presence of multiple organ dysfunction syndrome (92% v 25%). CONCLUSION: Both definitions identified similar groups of patients. The incidence in this population was higher than that reported elsewhere, but mortality and cause were similar to those in developed countries. Poor outcome was associated with sepsis, a high admission PRISM score, and simultaneous occurrence of other organ dysfunction. PMID- 9875024 TI - Metaphyseal growth arrest lines in psychosocial short stature. AB - Metaphyseal growth arrest lines are seen in children who experience significant physical stress such as infection or malnutrition over a sufficient period of time. These lines have not been reported previously in children with psychosocial short stature (PSS). Two boys and a girl with PSS with metaphyseal growth arrest lines on skeletal radiographs at the time of maximal stress in their homes are described. All three had reversible growth hormone insufficiency during admission, which is pathognomic for PSS. Multiple growth arrest lines in the distal end of the radius or vertebrae should alert clinicians to an alternative diagnosis in a child with growth hormone insufficiency. This may provide a clue to the diagnosis of occult PSS. PMID- 9875025 TI - Long-term outcome of brain manganese deposition in patients on home parenteral nutrition. AB - Manganese intoxication has been described in children on long term parenteral nutrition presenting with liver and nervous system disorders. Cases are reported of a brother and sister on long term parenteral nutrition with hypermanganesaemia and basal ganglia manganese deposition, detected by magnetic resonance imaging (MRI), without overt neurological signs. Following reduction of manganese intake, basal ganglia manganese was monitored by repeated MRI, and neurological and developmental examinations. An MRI intensity index of the globus pallidus declined over a three year period from 0.318 and 0.385 to 0.205 and 0.134 with concomitant falls in whole blood manganese from 323 and 516 to 226 and 209 nmol/l (normal range, 73-210 nmol/l). Unlike adult experience these children developed normally without neurological signs. In conclusion, deposited manganese is removed from neural tissue over time and the prognosis is good when neurological manifestations and liver disease are absent. PMID- 9875026 TI - Rectal biopsy in the investigation of constipation. AB - AIMS: To develop criteria to prevent unnecessary rectal biopsies in constipated children. METHODS: A retrospective review of 186 rectal biopsies from 141 children, comparing the age at onset of symptoms with the diagnosis of Hirschsprung's disease. RESULTS: All of the 17 children with Hirschsprung's disease had the onset of symptoms before the age of 4 weeks. Twenty seven children had delayed passage of meconium (more than 48 hours) of whom 10 had Hirschsprung's disease. Three children with Hirschsprung's disease were referred after the neonatal period (2 months, 11 months, and 3 years) but all had the onset of symptoms before 4 weeks of age. CONCLUSION: If the age at onset of constipation is after the neonatal period, a rectal biopsy is unnecessary. PMID- 9875027 TI - Seasonality of sudden infant death syndrome in mainland Britain and Ireland 1985 95. AB - By the end of 1995 four years had passed since the dramatic fall in the incidence of sudden infant death syndrome (SIDS), following the "back to sleep" campaign. This time lag permitted a more definitive epidemiological reassessment than had been possible before. The extent of seasonal variation from 1992-5 fell by about half, occurring mainly as a single step down in 1992. The fall was relatively greater in winter than in summer. Before 1992 the extent of seasonal variation (amplitude) was greater in those age > or = 5 months compared with those aged < or = 4 months. Since 1992 the falls in incidence and amplitude have been greater in the younger group, suggesting that SIDS deaths in younger and older babies may have different causes. PMID- 9875028 TI - Position paper on the use of botulinum toxin in cerebral palsy. UK Botulinum Toxin and Cerebral Palsy Working Party. PMID- 9875029 TI - Ethambutol in tuberculosis: time to reconsider? PMID- 9875031 TI - Skeletal dysplasias. PMID- 9875030 TI - Guidelines for managing acute gastroenteritis based on a systematic review of published research. PMID- 9875032 TI - The Nikolsky sign in staphylococcal scalded skin syndrome. PMID- 9875033 TI - Superantigen scalded skin syndromes. PMID- 9875034 TI - Raised cerebrospinal fluid glycerol concentration associated with use of dichloroacetate. PMID- 9875035 TI - Heavy caffeine consumption in pregnancy, smoking, and sudden infant death syndrome. PMID- 9875036 TI - The vitamin K debacle: cut the Gordian knot but first do no harm. PMID- 9875037 TI - Inferences for health provision from survival data in cystic fibrosis. PMID- 9875038 TI - A new mixed micellar preparation for oral vitamin K prophylaxis: randomised controlled comparison with an intramuscular formulation in breast fed infants. AB - OBJECTIVE: To compare a new oral preparation of vitamin K1 (Konakion MM) containing lecithin and glycocholic acid with a standard intramuscular (IM) preparation during the first 8 weeks of life in exclusively breast fed infants. METHODS: Infants were randomised at birth to the IM group (1 mg vitamin K) or the oral group (2 mg given at birth and repeated at 7 and 30 days of life). Prothrombin time (INR), plasma vitamin K1, and PIVKA II (undercarboxylated prothrombin) were monitored at 14, 30, and 56 days of age. RESULTS: Seventy nine infants were randomised to the oral group and 77 to the IM group. Sixty seven infants in each group completed eight weeks of the study. Prothrombin times did not differ between the two groups. Mean (SD) plasma vitamin K1 values (in ng/ml) decreased in both groups over time, but were higher in the oral group at 14 and 56 days: 2.0 (1.6) v 1.3 (1.1) at 14 days; 0.5 (0.3) v 0.5 (0.7) at 30 days; and 0.5 (0.8) v 0.2 (0.2) at 56 days of life. PIVKA II was raised (> or = 0.1 AU/ml) in cord blood in 47% of the infants. By 14 days, only one infant in each group had a raised PIVKA II value and both of these initially had high concentrations of PIVKA II in cord blood. At 30 days, there were no raised PIVKA II values. At 56 days, there were no raised PIVKA II values in the oral group, although three infants in the IM group had raised values. CONCLUSIONS: Plasma vitamin K concentrations were at least equal or significantly higher in babies given oral vitamin K supplements compared with IM treated babies at the time points measured. Through the first 8 weeks of life, multiple doses of the new oral preparation maintain haemostasis and vitamin K status in breast fed infants at least equal to that of the intramuscular preparation. PMID- 9875039 TI - Transition from school to adult life for physically disabled young people. AB - The transition process for young physically disabled people (n = 87) was studied in three districts to determine how it varied between schools, and how it was perceived by the disabled and their carers. The experiences of the young people were compared with guidelines for good practice based on relevant legislation. Results showed that young disabled people experienced a poor handover to adult services if they had no "statement of special educational need" or if they went to further education college. Young people with cerebral palsy or complex multiple problems fared less well than those with spina bifida or juvenile chronic arthritis. There are several deficiencies in the legislation. Physically disabled young people should receive a transition review regardless of whether they have a "statement". The range of severity and type of disorders among the young physically disabled argues for a range of services--the category is too broad to be useful for research, service planning, and provision. PMID- 9875040 TI - Mothers' reports of infant crying and soothing in a multicultural population. AB - OBJECTIVES: To investigate the prevalence of infant crying and maternal soothing techniques in relation to ethnic origin and other sociodemographic variables. DESIGN: A questionnaire survey among mothers of 2-3 month old infants registered at six child health clinics in Amsterdam, the Netherlands. SUBJECTS: A questionnaire on sociodemographic characteristics and crying behaviour was completed for 1826 of 2180 (84%) infants invited with their parents to visit the child health clinics. A questionnaire on soothing techniques was also filled out at home for 1142 (63%) of these infants. RESULTS: Overall prevalences of "crying for three or more hours/24 hour day" "crying a lot", and "difficult to comfort" were 7.6%, 14.0%, and 10.3%, respectively. Problematic infant crying was reported by 20.3% of the mothers. Of these infants, only 14% met all three inclusion criteria. Problematic crying occurred less frequently among girls, second and later born children, Surinamese infants, and breast fed infants. Many mothers used soothing techniques that could affect their infant's health negatively. Shaking, slapping, and putting the baby to sleep in a prone position were more common among non-Dutch (especially Turkish) mothers than among Dutch mothers. Poorly educated mothers slapped their baby more often than highly educated mothers. CONCLUSIONS: Mothers' reports of infant crying and soothing varied sociodemographically. Much harm may be prevented by counselling parents (especially immigrants) on how and how not to respond to infant crying. Health education should start before the child's birth, because certain soothing techniques could be fatal, even when practised for the first time. PMID- 9875041 TI - Growth monitoring: testing the new guidelines. AB - OBJECTIVE: To assess the impact of recent guidelines from the UK joint working party of child health surveillance recommending that all children be measured at age 5 and again between 7 and 9 years of age to determine how many normal school age children are likely to be referred for specialist assessment. METHODS: The longitudinal data of 486 children measured by school nurses in a community setting were examined and compared with measurements made in a research setting by a single, skilled observer. MAIN OUTCOME MEASURES: Number of children identified as having abnormal stature (< 0.4th or > 99.6th centile) and abnormal growth rate height standard deviation score (HSDS) change > 0.67). RESULTS: The community survey identified seven (1.4%) children as having abnormal stature (four short, three tall), 11 (2.3%) were identified as "slow growing", and nine (1.9%) increased their HSDS by more than 0.67. These results were comparable to data collected in ideal research conditions. CONCLUSIONS: Following the recommendations would not result in an excess number of inappropriate referrals. However, this study highlights several unresolved issues such as interobserver variability and time interval between measurements. A large scale prospective study should be considered to establish realistic and cost-effective criteria before implementation of a national screening programme. PMID- 9875042 TI - Efficacy of Zoladex LA (goserelin) in the treatment of girls with central precocious or early puberty. AB - OBJECTIVE: To assess the efficacy of a longer acting preparation of the gonadotrophin releasing hormone (GnRH) analogue goserelin (Zoladex LA, 10.8 mg) in 12 girls with central precocious or early puberty. METHODS: Two girls started treatment de novo; the remainder had been on suppressive treatment for a median duration of 1.5 (range, 0.2-5.6) years. Assessment comprising auxology, pubertal staging, and pelvic ultrasound examination was carried out at weeks 0, 4, 8, 10, and 12 (first cycle) and weeks 8, 10, and 12 (second cycle) to evaluate the required injection frequency. Thereafter, assessment was performed on the day of injection. Zoladex LA was given every 12 weeks unless pubertal progression occurred. RESULTS: Satisfactory control was achieved in eight patients using this regimen, and three patients required more frequent injections. One girl was removed from the study because of clinical progression and extreme mood swings. No serious adverse effects occurred. Mean height velocity during the study period was 4.5 cm/year (range, 3.1-6.6) compared with 6.5 cm/year (range, 3.8-9.6) before treatment in nine patients for whom data were available. CONCLUSIONS: Zoladex LA was effective in controlling precocious puberty in girls when given at intervals of 9-12 weeks and it is recommended that an initial assessment is made eight weeks after beginning treatment. PMID- 9875043 TI - Cross sectional study of the relation between sibling number and asthma, hay fever, and eczema. AB - OBJECTIVES: To document the relation between sibling number and atopic disease, and to assess the contribution of possible confounding factors to the protective effect of siblings in relation to asthma and hay fever. DESIGN AND SUBJECTS: Cross sectional survey by parental questionnaire in Tasmania, Australia, on 6378 children (92% of those eligible) who reached 7 years of age during 1995. METHODS: Exercise challenge lung function testing was conducted on 428 children. Analyses reported were conducted on singleton births only (n = 6158). RESULTS: The prevalences of a history of asthma ever, hay fever, and eczema were 27%, 19%, and 22%, respectively. Asthma and hay fever, but not eczema, were inversely related to sibling number, with evidence of a dose-response trend. The mean age at onset for asthma or wheezy breathing decreased as the number of siblings increased. The inverse association between sibling number and asthma or hay fever persisted after adjustment for several confounders, such as parental smoking or breast feeding, but did not persist after adjustment for household size in 1995. CONCLUSIONS: The protective effect of high sibling number could not be separated from household size at age 7, and it appears to be operating after birth and influences the age at onset of asthma symptoms. Further work to increase knowledge of how the protective effect of the presence of siblings works might have important implications for the understanding of the pathogenesis of asthma. PMID- 9875044 TI - Long-term follow up of 69 patients treated for optic pathway tumours before the chemotherapy era. AB - AIM: To analyse the long-term results of conservative management with radiotherapy in patients with optic pathway tumours. DESIGN: All 69 patients were symptomatic at diagnosis and most neoplasms involved the optic chiasm and hypothalamus. RESULTS: At 10 years, overall survival and progression free survival were 83% and 65.5%, respectively. After radiotherapy, vision improved in 18 patients and remained stable in 29 other patients. Cerebrovascular complications occurred in nine of 53 patients treated with radiotherapy after a median interval of two and a half years. These complications were five times more frequent in patients with neurofibromatosis type 1 (NF1). Severe intellectual disabilities were present in 18 children, most of whom underwent irradiation at a very young age (median age, 4 years). IMPLICATIONS: Radiotherapy is a valuable treatment in terms of tumour response, visual outcome, and progression free survival. However, in young children and in patients with NF1, major sequelae are encountered and new treatment strategies should be proposed for these patients. PMID- 9875045 TI - Colonic wall thickness, pancreatic enzyme dose and type of preparation in cystic fibrosis. AB - Increased colonic wall thickness has been reported in patients exposed to large doses of high strength pancreatic enzyme preparations who did not develop fibrosing colonopathy. This has been interpreted as evidence for a spectrum of subclinical disease. The relation between sonographically measured colonic wall thickness and pancreatic enzyme preparation and dose was studied in 86 children with cystic fibrosis (CF). Colonic wall thickness of a control group was also measured. The average thickness in all colonic regions was higher in the CF group (overall average range 0.7-2.5 mm v 0.6-1.4 mm in the control group). There was no significant relation between colonic wall thickness and age, sex, total dose of lipase, or copolymer. Apart from one patient with an early colonic stricture, none of those exposed to high doses of lipase, or the methacrylic acid copolymer Eudragit L30 D55, showed evidence of subclinical damage to the colon. The reproducibility of the sonographic measurements was poor. PMID- 9875046 TI - Lower urinary tract dysfunction in children with central nervous system tumours. AB - The findings in 10 children with neuropathic vesicourethral dysfunction after the onset of a central nervous system tumour are presented. Eight had a spinal tumour and two a brainstem tumour. Bladder dysfunction occurred late in most children except in those with neoplastic infiltration of the conus and cauda equina. Moreover, tumour recurrence was often heralded by loss of bladder control before other neurological signs became obvious. Videourodynamics (VUD) showed various combinations of "filling" and "voiding" dysfunction in tumours extending from the pons to the cauda equina, whereas an isolated "filling" dysfunction was evident in the patient with a suprapontine tumour. Urinary incontinence and recurrent urine infection can be immensely distressing to children and their families, particularly when they have had to cope with the stress of diagnosis and treatment of the underlying tumour. Different management strategies, based on VUD findings, are discussed highlighting the impact these have on the children's quality of life. PMID- 9875047 TI - A population study of chromosome 22q11 deletions in infancy. AB - AIMS: To determine the prevalence of submicroscopic deletions within chromosome band 22q11 in infants with significant heart disease and compare this with the prevalence of other chromosomal abnormalities causing significant heart disease. To determine a minimum prevalence of deletions within chromosome band 22q11 in infants in the general population. METHODS: Chromosome analysis was performed on samples from infants born in the former UK Northern Health Region in 1994 and 1995 who either had significant heart disease or who were suspected to have a chromosome band 22q11 deletion following referral to the Northern Genetics Service. Significant heart disease was defined as major structural malformation or cases where invasive investigation or intervention was required in infancy. RESULTS: Chromosome band 22q11 deletions were identified in nine infants in a population of 69,129 livebirths, giving a minimum prevalence of 13 per 100,000 (95% confidence interval 4.5 to 21.5). Six cases had significant heart disease, one of whom died before diagnosis. In the same population there were 53 cases of trisomy 21, 15 of whom had significant heart disease. CONCLUSION: The most common chromosomal cause of significant congenital heart disease remains trisomy 21, while the second most common chromosomal cause is deletion in chromosome band 22q11. PMID- 9875048 TI - Audiological abnormalities in the Klippel-Feil syndrome. AB - Klippel-Feil syndrome (KFS) is defined as a short neck with decreased movement and low posterior hairline. Radiologically, there is a failure of cervical segmentation. Deafness is a well known associated feature and may be of sensorineural, conductive, or mixed type. The audiological assessment of 44 patients with KFS is reported. Thirty five were found to have abnormalities on audiological testing. The presence of other features in these patients, reviews of data from other reports, and the need for further study are discussed. PMID- 9875050 TI - Intelligence and development in Aarskog syndrome. AB - AIM: To test the hypothesis that overall intelligence quotient (IQ) is decreased in patients with Aarskog syndrome. METHODS: 21 boys under 17 years of age with a confirmed clinical diagnosis of Aarskog syndrome were assessed using the Griffiths mental development scales and the British ability scales. RESULTS: IQ ranged from 68 to 128 and followed a normal distribution. CONCLUSION: This study does not support the hypothesis that Aarskog syndrome is associated with a lowering of mean IQ. PMID- 9875049 TI - Sexually transmitted organisms in sexually abused children. AB - OBJECTIVE: To establish the prevalence of sexually transmitted organisms and other genital organisms in potentially sexually abused children. DESIGN: Prospective study of children attending an inner London department of community paediatrics for evaluation of possible sexual abuse. SUBJECTS: Children under 16 referred for evaluation of possible sexual abuse. OUTCOME MEASURES: Prevalence of sexually transmitted organisms in relation to age, symptoms, and type of abuse. RESULTS: Swabs were taken from 159 of 242 girls evaluated. The overall prevalence of sexually transmitted organisms was 3.7%: three girls were infected with gonorrhoea, four with Trichomonas vaginalis, and two with Chlamydia trachomatis. One girl had all three infections plus mycoplasmas. Mycoplasmas were identified in 22% of girls swabbed. Of 30 boys swabbed, none yielded a sexually transmitted organism. CONCLUSIONS: There is a low prevalence of definitely sexually transmitted organisms in children who might have been abused. Other organisms possibly associated with sexual activity can be identified in this population. Screening for infection should be mandatory in presumed sexually abused girls with vaginal discharge and ideally should be undertaken in all children attending for evaluation of sexual abuse. PMID- 9875051 TI - Complications of modified Blalock-Taussig shunts mimicking pulmonary disease. AB - The modified Blalock-Taussig shunt is commonly performed as early palliation in cyanotic congenital heart disease. To highlight unusual diagnostic problems in such patients, two cases are reported in whom shunt complications were initially diagnosed as lobar pneumonia and tuberculosis, respectively. The children, an 8 month old boy and a 3 year old boy, had false aneurysm secondary to infection of the graft and dilatation of the left pulmonary artery caused by blood flow through the shunt, respectively. PMID- 9875052 TI - Hyperammonaemic encephalopathy after a subureteric injection for vesicoureteric reflux. AB - A 6 year old boy developed hyperammonaemic encephalopathy following a subureteric injection for treatment of vesicoureteric reflux. The hyperammonaemia may be explained by a postoperative urinary tract infection with a urea splitting organism, leading to raised urine ammonia that was absorbed easily across a dilated urinary tract. Agitation and alteration in consciousness level following a urological procedure, in a child with a dilated urinary tract, may be signs of a treatable hyperammonaemic encephalopathy. PMID- 9875053 TI - Maximising the effectiveness of undergraduate teaching in the clinical setting. PMID- 9875055 TI - How I manage chronic fatigue syndrome. PMID- 9875054 TI - Chronic fatigue syndrome. PMID- 9875056 TI - Preclinical diagnosis of abdominal tumours by ultrasound examination. PMID- 9875057 TI - Developing injury surveillance systems in accident and emergency departments. PMID- 9875058 TI - Serum bicarbonate and the severity of dehydration in gastroenteritis. PMID- 9875059 TI - Physical activity and metabolic cardiovascular syndrome. PMID- 9875060 TI - Energy requirements assessed using the doubly-labelled water method. PMID- 9875061 TI - The assembly of triacylglycerol-rich lipoproteins: an essential role for the microsomal triacylglycerol transfer protein. AB - Raised plasma triacylglycerol is an independent risk factor for cardiovascular disease, and an understanding of factors which regulate the synthesis and degradation of lipoproteins which carry triacylglycerol in the blood may lead to novel approaches to the treatment of hypertriacylglycerolaemia. An active microsomal triacylglycerol transfer protein (MTP) is essential for the assembly of particles which transport triacylglycerol through the circulation. After absorption in the intestine, dietary fat and fat-soluble vitamins are incorporated into chylomicrons in the intestinal epithelial cells, and these lipoproteins reach the bloodstream via the lymphatic system. Patients with the rare genetic disorder, abetalipoproteinaemia, in which MTP activity is absent, present clinically with fat-soluble vitamin and essential fatty acid deficiency, indicating a key role for MTP in the movement of fat into the body. The triacylglycerol-rich lipoprotein found in fasting blood, VLDL, is assembled in the liver by an MTP-dependent process similar to chylomicron assembly, and transports triacylglycerol to extra-hepatic tissues such as adipose tissue and heart. In the absence of MTP activity, VLDL are not synthesized and only extremely low levels of triacylglycerol are present in the blood. Dietary components, including fat, cholesterol and ethanol, can modify the expression of the MTP gene and, hence, MTP activity. The present review summarizes current knowledge of the role of MTP in the assembly and secretion of triacylglycerol rich lipoproteins, and the regulation of its activity in both animal and cell systems. PMID- 9875062 TI - Effects of the bacterial status of rats on the changes in some liver cytochrome P450 (EC 1.14.14.1) apoproteins consequent to a glucosinolate-rich diet. AB - The aim of the present work was to investigate the influence of the intestinal microflora on the changes in hepatic cytochrome P450 apoproteins induced by dietary glucosinolates. Ten rats harbouring a conventional digestive microflora were offered either a diet containing 390 g myrosinase-free rapeseed meal/kg (n 5) or a control diet devoid of glucosinolates (n 5). A similar trial was performed using germ-free rats. After 4 weeks of exposure to the dietary regimens, animals were slaughtered and their livers removed for preparation of microsomes and analysis of cytochrome P450 (EC 1.14.14.1). The glucosinolate-rich diet decreased the concentration of total cytochrome P450 in conventional rats only (-34%). The bacterial status did not modify the concentration of apoproteins CYP1A2 and CYP2B1/B2, but greatly decreased the concentration of the male constitutive isoform CYP2C11 (-53 and -45% respectively in conventional and germ free rats). Germ-free rats fed on the glucosinolate-rich diet had a greater concentration of CYP3A (+139%) and a lower concentration of CYP2E1 (-32%) than their counterparts fed on the control diet. However, these differences were absent in conventional animals. On the whole, the influence of the intestinal microflora on the changes in hepatic cytochrome P450 due to the consumption of cruciferous vegetables is very complex and obviously involves different mechanisms according to the apoprotein. PMID- 9875063 TI - A quantitative analysis of the relationship between habitual energy expenditure, fitness and the metabolic cardiovascular syndrome. AB - Previous epidemiological studies have suggested an association between low levels of physical activity, fitness and the metabolic cardiovascular syndrome. However, many studies have used subjective non-quantitative questionnaire-based methods for assessing physical activity which do not distinguish between the different dimensions of this complex exposure, and in which measurement error in the exposure has not been estimated. These deficiencies in the measurement of this exposure complicate the interpretation of the results of epidemiological studies, and consequently make it difficult to design appropriate interventions and to estimate the expected benefit which would result from intervention. In particular, it is unclear whether public health advice should be to increase total energy expenditure, or to attempt to raise fitness by recommending periods of vigorous activity. To separate the effects of fitness and total energy expenditure in the aetiology of the metabolic cardiovascular syndrome, we measured the physical activity level (PAL), defined as total energy expenditure: BMR, and fitness (maximum O2 consumption (VO2max per kg), measured in a sub maximal test) in a cross-sectional population-based study of 162 adults aged 30 40 years. Heart-rate monitoring with individual calibration was used to measure total energy expenditure using the HRFlex method (Ceesay et al. 1989) which has been validated previously against doubly-labelled water and whole-body calorimetry. The relationship between a single measure of PAL, VO2max per kg and the usual or habitual level for each exposure was measured in a sub-study of twenty-two subjects who undertook four repeated measures over the course of 1 year. This study design allows the reliability coefficient to be computed, which is used to adjust the observed associations for measurement error in the exposure. Twelve men (16.4%) and sixteen women (18.0%) were defined as having one or more features of the metabolic cardiovascular syndrome. The univariate odds ratio for each increasing quartile for PAL was 0.64 (95% CI 0.43-0.94) and was 0.49 (95% CI 0.32-0.74) for VO2max per kg, suggesting that the association with the metabolic cardiovascular syndrome was stronger for fitness than for PAL. However, after adjustment for obesity and sex, and correction for exposure measurement error, the odds ratio per quartile for PAL was 0.32 (95% CI 0.13 0.83) and 0.44 (95% CI 0.24-0.78) for VO2max per kg. Thus, although univariate analysis would suggest that fitness has a stronger association with the metabolic cardiovascular syndrome than PAL, this conclusion is reversed once confounding and the differences in measurement error are considered. We conclude from the present study that the metabolic cardiovascular syndrome is strongly associated with reduced habitual energy expenditure. The method employed to assess the exposure in the present study demonstrates the utility of assessing a known dimension of physical activity using a physiologically-based and objective measure with repeated estimation to adjust for measurement error. Such quantitative epidemiological data provide the basis for planning and evaluating the expected benefit of population-level interventions. PMID- 9875064 TI - Effect on plasma lipids and lipoproteins of replacing partially hydrogenated fish oil with vegetable fat in margarine. AB - We have compared the effects on lipoproteins and haemostatic variables of two hard margarines with similar functional properties, one traditional margarine containing partially hydrogenated fish oil (PHFO), and one experimental margarine based on vegetable oil (VO). Both were all-purpose cooking margarines with nearly identical functional properties. Trans fatty acids from PHFO in the traditional margarine were replaced mostly by saturated, monounsaturated and trans fatty acids of vegetable origin in the new formulation. Both test margarines contained approximately the same amount of cis polyunsaturated fatty acids. Sixteen female normolipidaemic students consumed each diet with the two test margarines for 14 d in random order (crossover design). The amount of fat was 31% energy in the PHFO diet and 32% energy in the VO diet. The test margarines provided approximately 26% energy in both diets. In the PHFO diet 7.8% of the energy was derived from trans fatty acids and 9.2% from saturated fatty acids (12:0, 14:0 and 16:0) while in the VO diet, 1.1% energy was derived from trans fatty acids and 13.3% from saturated fatty acids (12:0, 14:0 and 16:0). The natural content of cholesterol in PHFO was deliberately not balanced by addition of cholesterol to the VO diet, thus the PHFO diet contained 215 mg and the VO diet 86 mg cholesterol per 8.5 MJ. LDL-cholesterol concentration was 19% higher in subjects on the PHFO diet compared with the VO diet (P < 0.01). The ratio LDL-cholesterol:HDL-cholesterol was 12.6% higher in subjects on the PHFO diet compared with the VO diet (P < 0.01). The level of apolipoprotein (apo)A-I was 6% lower in subjects on the PHFO diet compared with the VO diet (P < 0.01). The ratio apoB:apoA-I was 10.4% higher in subjects on the PHFO diet than on the VO diet (P < 0.01). There were no significant differences in total cholesterol, HDL-cholesterol, triacylglycerols, apoB, lipoprotein(a) and haemostatic variables between the diets. Our results demonstrate that PHFO, with its unfavourable effects on plasma lipids, can be replaced by vegetable oils in margarine without appreciable loss of functional properties but with significant improvement in the effects on plasma lipoproteins. PMID- 9875065 TI - Fermentation of non-starch polysaccharides in mixed diets and single fibre sources: comparative studies in human subjects and in vitro. AB - The present study investigated whether the extent of fermentation of NSP in human subjects could be predicted by an in vitro batch system. Fibre sources studied were five mixed diets containing different amounts and types of fibre and three single fibre sources (citrus fibre concentrate, coarse and fine wholemeal rye bread). Fermentation in human subjects was determined in balance experiments in women who were also donors of the faecal inocula. In vitro fermentations were performed with fibre residues prepared from duplicates of the fibre-containing foods consumed during the balance trials. Fermentation of total NSP in vivo was between 65.8 and 88.6% for the mixed diets and 54.4, 58.0 and 96.9% for the coarse and fine wholemeal rye breads and the citrus fibre concentrate respectively. For the mixed diets and the citrus fibre concentrate, mean differences between the extent of NSP degradation after 24 h in vitro incubation and that in vivo were between -0.7 and 5.0%. Differences were significant for one diet (P < 0.05). For the wholemeal rye breads, the fermentation in vitro exceeded that in vivo significantly, but the magnitude of the difference in each case was small and without physiological importance. Particle size of breads had no influence on the extent of NSP degradation. These results indicate that the in vitro batch system used could provide quantitative data on the fermentation in vivo of NSP in mixed diets and some single fibre sources. An in vitro incubation time of 24 h was sufficient to mimic the NSP degradation in vivo. PMID- 9875066 TI - Estimation of energy expenditure in free-living red deer (Cervus elaphus) with the doubly-labelled water method. AB - Energy expenditure was estimated using the doubly-labelled water (DLW) method in summer in five free-living adult, non-pregnant, non-lactating, red deer (Cervus elaphus) hinds (weight 107.3 (SE 0.9) kg; age 6 (SE 1) years) on lowland pasture under typical farming conditions. Climatic conditions were monitored throughout the experiment. Errors due to 2H losses in CH4 and faeces were calculated from previous estimates of stoichiometries. CH4 production, fractionated water loss, urinary N and O2 consumption were estimated using an iterative approach. The water flux (rH2O) in these animals consuming only fresh grass was 12 (SE 0.5) kg/d, the CO2 production (rCO2) was 1271 (SE 40) litres/d and the mean energy expenditure was 25 (SE 0.8) MJ/d. There were no significant differences in the isotope distribution spaces and flux rates, rH2O, rCO2 or energy expenditure using the multi-point or two-point approaches to calculation. The DLW-derived energy expenditure of 25 MJ/d is approximately 20% higher than the recommended intake of 21 MJ/d for adult hinds kept outdoors (Adam, 1986) and, at 757 kJ/kg0.75 per d, one third higher than the value of 570 kJ/kg0.75 per d for stags penned indoors (Key et al. 1984). PMID- 9875067 TI - Effect of feeding tuna oil or soyabean oil as supplements to sows in late pregnancy on piglet tissue composition and viability. AB - To investigate whether long-chain n-3 polyunsaturated fatty acids could cross the porcine placenta in late pregnancy and alter neonatal piglet tissue composition, multiparous sows (seven per diet) were fed on diets containing a supplement (30 g/kg) of either soyabean oil or tuna oil for the last 21 d of pregnancy and the first 7 d of lactation. The proportions of all fatty acids, except 18:1n-7, differed between diets: in particular, the tuna-oil-containing diet supplied more 22:6n-3 and less 18:2n-6 fatty acids than the soyabean-oil-containing diet. The proportions of n-3 fatty acids, particularly 22:6n-3 (g/100 g total fatty acids) in sow plasma, colostrum and milk were increased and the proportion of 18:2n-6 was decreased by feeding tuna oil. Piglet tissue n-3 fatty acid proportions (particularly 22:6n-3), obtained shortly after birth, were increased in piglets born to tuna-oil-fed sows compared with progeny of soyabean-oil-fed sows. The increase in the proportion of n-3 fatty acids (g/100 g total fatty acids) in piglet tissues as a result of tuna-oil feeding, compared with soyabean-oil feeding, was in the order plasma > liver > erythrocytes > spleen > brain > retina. Piglets born to tuna-oil-fed sows had a lower viability score at birth than the progeny of soyabean-oil-fed sows. The proportions of long-chain n-3 fatty acid in tissues of new-born piglets were increased by feeding tuna oil to the sow in late pregnancy; however no improvements in piglet viability were observed. PMID- 9875068 TI - Dietary fish oil and digestible protein modify susceptibility to lipid peroxidation in the muscle of rainbow trout (Oncorhynchus mykiss) and sea bass (Dicentrarchus labrax). AB - The effects of dietary fish oil and digestible protein (DP) levels on muscle fatty acid composition and susceptibility to lipid peroxidation were studied in two representative fish species for human nutrition, from fresh and seawater, rainbow trout (Oncorhynchus mykiss) and European sea bass (Dicentrarchus labrax). In rainbow trout, higher concentrations of dietary fat and DP led to higher weight gain (g/d) (P = 0.001 and P = 0.043 respectively). Additionally, an interaction effect was observed in this species, since the effect of DP was only evident when the dietary fat concentration was low (P = 0.043). A similar tendency was also observed in European sea bass, although with less marked differences among nutritional treatments. Trout fed on diets with a higher concentration of dietary fat had higher concentrations of intramuscular total and neutral lipids in the dorsal muscle (P = 0.005). Increased levels of dietary DP led to significantly lower concentrations of polar lipids in the dorsal muscle of both rainbow trout (P = 0.005) and European sea bass (P = 0.006). In the neutral fraction of intramuscular lipids of dorsal muscle the concentration of n-3 fatty acids was positively affected by the dietary fat concentration in both rainbow trout (P = 0.04) and sea bass (P = 0.001). Muscle homogenates from trout and sea bass fed on diets rich in fish oil showed a significantly higher susceptibility to oxidation than muscle homogenates from fish fed on low-fat diets (P = 0.001). The higher DP concentration also increased susceptibility to oxidation. Moreover, in rainbow trout an interaction effect was observed where the pro-oxidant effect was of higher magnitude when the dietary concentration of both nutrients, fat and protein, was high (P = 0.004). PMID- 9875069 TI - Protein deficiency during pregnancy and lactation impairs glucose-induced insulin secretion but increases the sensitivity to insulin in weaned rats. AB - We studied glucose homeostasis in rat pups from dams fed on a normal-protein (170 g/kg) (NP) diet or a diet containing 60 g protein/kg (LP) during fetal life and the suckling period. At birth, total serum protein, serum albumin and serum insulin levels were similar in both groups. However, body weight and serum glucose levels in LP rats were lower than those in NP rats. At the end of the suckling period (28 d of age), total serum protein, serum albumin and serum insulin were significantly lower and the liver glycogen and serum free fatty acid levels were significantly higher in LP rats compared with NP rats. Although the fasting serum glucose level was similar in both groups, the area under the blood glucose concentration curve after a glucose load was higher for NP rats (859 (SEM 58) mmol/l per 120 min for NP rats v. 607 (SEM 52) mmol/l per 120 min for LP rats; P < 0.005). The mean post-glucose increase in insulin was higher for NP rats (30 (SEM 4.7) nmol/l per 120 min for NP rats v. 17 (SEM 3.9) nmol/l per 120 min for LP rats; P < 0.05). The glucose disappearance rate for NP rats (0.7 (SEM 0.1) %/min) was lower than that for LP rats (1.6 (SEM 0.2) %/min; P < 0.001). Insulin secretion from isolated islets (1 h incubation) in response to 16.7 mmol glucose/l was augmented 14-fold in NP rats but only 2.6-fold in LP rats compared with the respective basal secretion (2.8 mmol/l; P < 0.001). These results indicate that in vivo as well as in vitro insulin secretion in pups from dams maintained on a LP diet is reduced. This defect may be counteracted by an increase in the sensitivity of target tissues to insulin. PMID- 9875070 TI - Dietary lipids and evolution of the human brain. PMID- 9875071 TI - Dietary lipids and evolution of the human brain. PMID- 9875072 TI - Transfusion associated graft versus host disease and its prevention. PMID- 9875073 TI - Heart rate variability and cardiac failure. PMID- 9875074 TI - Of bombers, radiologists, and cardiologists: time to ROC. PMID- 9875075 TI - Congresses. PMID- 9875076 TI - Myocardial hibernation and stunning: from physiological principles to clinical practice. PMID- 9875077 TI - Troponin T measurement after myocardial infarction can identify left ventricular ejection of less than 40%. AB - AIM: To determine whether measurement of serum troponin T concentration after first acute myocardial infarction can be used to identify patients with a left ventricular ejection fraction of < 40%, who have an adverse prognosis. METHODS: Troponin T concentration was measured, and coronary and left ventriculography performed in 50 consecutive patients with acute myocardial infarction. Angiographic left ventricular ejection fraction was compared with serum troponin T concentration. Patients with previous myocardial infarction were excluded. RESULTS: There was a strong negative correlation between left ventricular ejection fraction and troponin T concentration. Spearman's rank correlation coefficient (corrected for ties) was -0.72 (95% confidence intervals (CI) -0.55 to -0.83; p < 0.0001). Analysis by receiver operator characteristic curve produced an area under the curve of 0.9773 (95% CI 0.9409 to 1.0136). A troponin T concentration of > 2.8 micrograms/l predicted a left ventricular ejection fraction of < 40% with a sensitivity of 100% (CI 84.6 to 100.0) and specificity of 92.9% (CI 76.5 to 99.1). Exclusion of patients who did not receive thrombolytic treatment did not significantly affect the results. CONCLUSION: Serum troponin T concentration measured 12-48 hours after admission for first myocardial infarction is a reliable, simple, quick, inexpensive, non-invasive method for identifying patients with a left ventricular ejection fraction of < 40% for whom there is a poor prognosis. PMID- 9875078 TI - Cardiac troponin T does not increase after electrical cardioversion for atrial fibrillation or atrial flutter. AB - OBJECTIVE: To determine whether cardiac troponin T increases after electrical cardioversion in patients with atrial fibrillation or atrial flutter. DESIGN: Serum creatine kinase (CK), creatine kinase-MB (CKMB), and cardiac troponin T were measured before, 24 hours, and 48 hours after cardioversion in 15 patients with atrial fibrillation or atrial flutter. RESULTS: 12 of the 15 patients (80%) were successfully cardioverted to sinus rhythm. The median number of shocks was three (range one to six), the median cumulative energy 710 J (50 to 1430 J), and the median peak energy 300 J (50 to 360 J). Total CK increased from a baseline median concentration of 92 (45 to 259) to 1324 (96 to 6660) U/l at 24 hours and 1529 (120 to 4774) U/l at 48 hours after cardioversion. There was a small increase in CKMB but the ratio of CKMB to CK did not increase. There was no increase in cardiac troponin T in any patient. CONCLUSIONS: Following electrical cardioversion of atrial fibrillation or atrial flutter, cardiac troponin T remains unchanged despite a large rise in total CK, indicating that the CK is derived from skeletal muscle and that myocardial injury does not occur. If cardiac troponin T is increased after cardioversion for atrial arrhythmias then other causes of myocardial damage should be sought. PMID- 9875079 TI - Direct current cardioversion does not cause cardiac damage: evidence from cardiac troponin T estimation. AB - AIM: To determine whether elective direct current (dc) cardioversion of atrial fibrillation/flutter causes myocardial damage. METHODS AND RESULTS: Cardiac troponin T and creatine kinase were estimated 20-28 hours after dc cardioversion in 51 patients who received dc shocks for elective cardioversion of chronic atrial fibrillation/flutter. Although creatine kinase was raised in 44 patients, cardiac troponin T was undetectable in all patients. CONCLUSION: Cardiac damage does not occur as a result of cardioversion. PMID- 9875080 TI - Audit of prehospital thrombolysis by general practitioners in peripheral practices in Grampian. AB - BACKGROUND: In the Grampian region early anistreplase trial (GREAT), domiciliary thrombolysis by general practitioners was associated with a halving of one year mortality compared with hospital administration. However, after completion of the trial and publication of the results, the use of this treatment by general practitioners declined sharply. OBJECTIVE: To increase the proportion of eligible patients receiving timely thrombolytic treatment from their general practitioners. SETTING: Practices in Grampian located > or = 30 minutes' travelling time from Aberdeen Royal Infirmary, where patients with suspected acute myocardial infarction were referred after being seen by general practitioners. AUDIT STANDARD: A call-to-needle time of 90 minutes, as proposed by the British Heart Foundation (BHF). METHODS: Findings of this audit of pre hospital management of acute myocardial infarction were periodically fed back to the participating doctors, when practice case reviews were also conducted. RESULTS: Of 414 administrations of thrombolytic treatment, 146 (35%) were given by general practitioners and 268 (65%) were deferred until after hospital admission. Median call-to-needle times were 45 (94% < or = 90) and 145 (7% < or = 90) minutes, respectively. Survival at one year was improved with prehospital compared with hospital thrombolysis (83% v 73%; p < 0.05). The proportion of patients receiving thrombolytic treatment from their general practitioners did not increase during the audit. CONCLUSIONS: In practices > or = 30 minutes from hospital, the BHF audit standard was readily achieved if general practitioners gave thrombolytic treatment, but not otherwise. Knowledge of the benefits of early thrombolysis, and feedback of audit results, did not lead to increased prehospital thrombolytic use. Additional incentives are required if general practitioners are to give thrombolytic treatment. PMID- 9875081 TI - Fibrinolytic potential is significantly increased by oestrogen treatment in postmenopausal women with mild dyslipidaemia. AB - OBJECTIVE: To study the effects of oestrogen replacement treatment on fibrinolytic potential in postmenopausal women. DESIGN: Randomised, double blind, placebo controlled trial of oral 17 beta-oestradiol. SETTING: Subjects were evaluated in the outpatient setting. PATIENTS: Nineteen postmenopausal women with mild dyslipidaemia, aged 44 to 69 years (mean (SD) 55.7 (6.7)). MAIN OUTCOME MEASURES: Fibrinolytic activity (fibrin plate assay) and tissue plasminogen activator (t-PA) antigen were measured at baseline and after three, six, and nine weeks of each treatment. RESULTS: After nine weeks of 2 mg oestradiol treatment, there was a significant increase in fibrinolytic potential compared with placebo, as indicated by an increase in fibrinolytic activity (mean (SEM), 80 (9) v 54 (5) mm2 of lysis in the fibrin plate, 2 mg v placebo, p = 0.002) and a decrease in t PA antigen (5.8 (0.9) v 8.4 (1.2) ng/ml, 2 mg v placebo, p < 0.001). There was a similar trend with the 1 mg dose but the changes were less noticeable. CONCLUSIONS: Hormone replacement treatment with 17 beta-oestradiol for nine weeks significantly increased fibrinolytic potential in postmenopausal women with mild dyslipidaemia. This suggests that the cardioprotective effect of oestrogen may be mediated, in part, by an increase in fibrinolytic potential. PMID- 9875082 TI - Late complications following permanent pacemaker implantation or elective unit replacement. AB - OBJECTIVE: To determine the rate of late complications following first implantation or elective unit replacement of a permanent pacemaker system. DESIGN: Analysis of pacemaker data and complications prospectively acquired on a computerised database. Complications were studied over an 11 year period from January 1984 to December 1994. SETTING: Tertiary referral cardiothoracic centre. PATIENTS: Records of 2621 patients were analysed retrospectively. MAIN OUTCOME MEASURES: Complications requiring repeat procedures occurring more than six weeks after pacemaker implantation or elective unit replacement. RESULTS: The overall rate of late complications was significantly lower after first implantation of a permanent pacemaker (34 cases, complication rate 1.4%, 95% confidence interval 0.9% to 1.9%) than after elective unit replacement (16 cases, complication rate 6.5% (3.3% to 9.7%). There were 20 cases of erosion, 18 infections, five electrode problems, and seven miscellaneous problems. Complications were more common with inexperienced operators (18.9% (6.0% to 31.8%)) than with experienced operators (0.9% (0.3% to 1.5%). CONCLUSIONS: The incidence of late complications following pacemaker implantation is low and compares favourably with early complication rates. The majority are caused by erosion and infection. Patients who have undergone elective unit replacement are at particular risk. PMID- 9875083 TI - Dispersion of regional wall motion abnormality in patients with long QT syndrome. AB - OBJECTIVE: To examine the left ventricular regional wall motion abnormality and to evaluate dispersion of this abnormality in patients with long QT syndrome. DESIGN: Left ventricular short axis images at basal and middle levels were recorded on videotape and digitised to reconstruct digitised M mode echocardiograms, from which left ventricular wall thickness curves were obtained. The wall thickening time (ThT) was defined as the period in which the instantaneous wall thickness exceeded 90% of the maximum wall thickness. ThT was measured at three segments in each of the septal and free wall sides of the left ventricle, a total of 12 segments. To examine the mechanical dispersion of the left ventricle, the difference between the maximum and minimum ThT of 12 segments in each subject was obtained. PATIENTS: Eight patients with congenital long QT syndrome (averaged QTc interval (SD) 509 (27) ms1/2) and 10 control subjects (QTc interval 397 (26) ms1/2) were examined. RESULTS: The averaged ThT values of the 12 segments pooled form all subjects were correlated with the QT intervals (r = 0.72, p < 0.005). Thus the averaged ThT in the long QT syndrome patients was longer than in the control subjects (p < 0.005). The segmental variation of ThT in the patients was greater than in the control subjects (p < 0.001). The dispersion of ThT in the patients was therefore larger than in control subjects (p < 0.005). However, the pattern of ThT variation in the patients varied according to the individual subject. CONCLUSIONS: There is not only electrical but also mechanical dispersion in the left ventricle of long QT syndrome patients. Regional assessment of ventricular wall motion may allow quantification of the spatial variation of wall motion abnormality. PMID- 9875084 TI - Prognostic significance of electrical alternans versus signal averaged electrocardiography in predicting the outcome of electrophysiological testing and arrhythmia-free survival. AB - OBJECTIVE: To investigate the accuracy of signal averaged electrocardiography (SAECG) and measurement of microvolt level T wave alternans as predictors of susceptibility to ventricular arrhythmias. DESIGN: Analysis of new data from a previously published prospective investigation. SETTING: Electrophysiology laboratory of a major referral hospital. PATIENTS AND INTERVENTIONS: 43 patients, not on class I or class III antiarrhythmic drug treatment, undergoing invasive electrophysiological testing had SAECG and T wave alternans measurements. The SAECG was considered positive in the presence of one (SAECG-I) or two (SAECG-II) of three standard criteria. T wave alternans was considered positive if the alternans ratio exceeded 3.0. MAIN OUTCOME MEASURES: Inducibility of sustained ventricular tachycardia or fibrillation during electrophysiological testing, and 20 month arrhythmia-free survival. RESULTS: The accuracy of T wave alternans in predicting the outcome of electrophysiological testing was 84% (p < 0.0001). Neither SAECG-I (accuracy 60%; p < 0.29) nor SAECG-II (accuracy 71%; p < 0.10) was a statistically significant predictor of electrophysiological testing. SAECG, T wave alternans, electrophysiological testing, and follow up data were available in 36 patients while not on class I or III antiarrhythmic agents. The accuracy of T wave alternans in predicting the outcome of arrhythmia-free survival was 86% (p < 0.030). Neither SAECG-I (accuracy 65%; p < 0.21) nor SAECG-II (accuracy 71%; p < 0.48) was a statistically significant predictor of arrhythmia-free survival. CONCLUSIONS: T wave alternans was a highly significant predictor of the outcome of electrophysiological testing and arrhythmia-free survival, while SAECG was not a statistically significant predictor. Although these results need to be confirmed in prospective clinical studies, they suggest that T wave alternans may serve as a non-invasive probe for screening high risk populations for malignant ventricular arrhythmias. PMID- 9875085 TI - Electrocardiographic changes after alcohol septal ablation in hypertrophic obstructive cardiomyopathy. AB - OBJECTIVE: To report acute and mid-term electrocardiographic changes in patients with hypertrophic obstructive cardiomyopathy (HOCM) after alcohol ablation of the first large septal branch of the left anterior descending coronary artery; and to relate electrocardiographic data with the left ventricular outflow tract pressure gradients. PATIENTS: Nine consecutive symptomatic patients with HOCM (mean (SD) age 45 (12) years). METHODS: Analysis of baseline and post-procedure ECGs and 24 hour ambulatory monitoring (up to six months). ECG data were related to left ventricular outflow tract pressure gradients. RESULTS: One patient developed complete atrioventricular block requiring permanent pacing. The PR interval was significantly prolonged up to third month after ablation. Immediately after the procedure all patients developed right bundle branch block. At the sixth month of follow up, right bundle branch block was present in four patients. New anterior ST elevation developed immediately after ablation in five of the nine patients, and new Q waves in four. The QRS duration was significantly prolonged immediately after ablation and during follow up. There was significant but transient prolongation of QT-mean and QTc-mean intervals. QT dispersion, QTc dispersion, and JTc-mean interval were not affected. JT and JTc dispersions were transiently prolonged. No serious ventricular arrhythmias were recorded during Holter monitoring, either before or after the procedure. There were no significant correlations between the left ventricular outflow tract pressure gradient and QTc, QT-d, QTc-d, JTc, JT-d, JTc-d, or QRS duration before and after ablation. CONCLUSIONS: Alcohol septal ablation for HOCM induces significant changes in the resting ECG in most patients, despite the occlusion of a relatively small artery. The changes include new Q waves, new bundle branch block, transient anterior ST segment elevation, atrioventricular block, and transient prolongation of QT interval. PMID- 9875086 TI - Sustained left ventricular diastolic dysfunction after exercise in patients with dilated cardiomyopathy. AB - OBJECTIVE: To investigate the recovery process of exercise induced diastolic dysfunction in heart failure, using Doppler echocardiographic techniques. DESIGN AND PATIENTS: Transmitral flow velocity profiles and standard noninvasive haemodynamic indices were obtained serially over seven days after symptom limited bicycle exercise tests in 18 patients with dilated cardiomyopathy and eight normal subjects. In three patients with cardiomyopathy we also measured the pulmonary capillary wedge pressure for 24 hours after exercise. RESULTS: The intensity of exercise, as assessed by respiratory gas analysis, was lower in patients with dilated cardiomyopathy than in normal subjects. Despite the higher exercise level, all haemodynamic variables returned to baseline within one hour after exercise in normal subjects. In contrast, patients with dilated cardiomyopathy showed a sustained decrease in the peak early diastolic filling velocity and a sustained increase in the deceleration time of early filling for 24 hours or more after exercise. Because other haemodynamic variables recovered within one hour after exercise even in patients with dilated cardiomyopathy, the postexercise changes in ventricular filling were not explained by changes in loading conditions. CONCLUSIONS: Exercise induced diastolic left ventricular dysfunction of the failing heart persists for 24 hours or more after exercise. The efficacy of exercise training on a daily basis in dilated cardiomyopathy requires further evaluation. PMID- 9875087 TI - Idiopathic dilated cardiomyopathy: lack of association with hepatitis C virus infection. AB - OBJECTIVE: To determine whether there is an association between hepatitis C virus (HCV) infection and dilated cardiomyopathy in a well defined area of north western Greece; such an association has been reported elsewhere. DESIGN: Evaluation of consecutive patients with chronic HCV infection for the presence of clinical or subclinical manifestations of dilated cardiomyopathy by history, physical examination, and non-invasive laboratory procedures (ECG, chest x ray, and echocardiography) before the initiation of interferon alpha treatment; investigation for HCV infection markers in patients with dilated cardiomyopathy by enzyme and immunoblot assays (antibodies to HCV) and the reverse transcriptase polymerase chain reaction (HCV RNA). SETTING: A tertiary referral centre for patients with chronic hepatitis and dilated cardiomyopathy. PATIENTS: 102 patients with well defined chronic HCV infection and 55 patients with well established dilated cardiomyopathy were evaluated. MAIN OUTCOME MEASURES: The need for HCV testing in patients with dilated cardiomyopathy, or follow up for heart disease in patients with chronic HCV infection. RESULTS: None of the patients with chronic HCV infection had clinical or subclinical evidence of dilated cardiomyopathy from history and laboratory findings. None of the patients with dilated cardiomyopathy was positive for antibodies to HCV or viraemic on HCV RNA testing. CONCLUSIONS: The study neither confirms the findings of other investigators, nor indicates a pathogenic link between HCV and dilated cardiomyopathy. For this reason, at least in Greece, testing for HCV in patients with dilated cardiomyopathy or follow up for heart disease in HCV patients appears unnecessary. Genetic or other factors could be the reason for this discrepancy if previously reported associations between HCV and dilated cardiomyopathy or hypertrophic cardiomyopathy were not coincidental. PMID- 9875088 TI - Clinical and morphological characteristics in Streptococcus bovis endocarditis: a comparison with other causative microorganisms in 177 cases. AB - AIM: To compare the clinical and morphological characteristics of patients with Streptococcus bovis endocarditis with those of patients with endocarditis caused by other microorganisms. METHODS: 177 consecutive patients (Streptococcus bovis, 22; other streptococci, 94; staphylococci, 44; other, 17) with definite infective endocarditis according to the Duke criteria were included. All patients underwent transthoracic and transoesophageal echocardiography. In 88 patients, findings from surgery/necropsy were obtained. RESULTS: S bovis endocarditis was associated with older patients, with a higher mortality (p = 0.04), and with a higher rate of cardiac surgery (p < 0.001) than other microorganisms, although embolic events were observed less often (p = 0.02). Pathological gastrointestinal lesions were detected in 45% of the patients. Multiple valves were affected in 68% of the patients with S bovis endocarditis and in 20% of those with other organisms (p < 0.001). Moderate or severe regurgitation occurred more often in S bovis endocarditis than with other microorganisms (p = 0.05). When surgery or necropsy was performed, infectious myocardial infiltration of the left ventricle was confirmed histopathologically in 36% of the patients with S bovis endocarditis and in 10% of those with other organisms (p = 0.002). CONCLUSIONS: S bovis endocarditis is a severe illness because of the more common involvement of multiple valves, and of the frequent occurrence of haemodynamically relevant valvar regurgitation and infectious myocardial infiltration. PMID- 9875089 TI - Reference ranges for cardiac dimensions and blood flow velocity in preterm infants. AB - AIM: To establish reference ranges for cardiac dimensions and Doppler measurements in preterm infants. METHODS: 79 infants of less than 34 weeks' gestation were examined by echocardiography on days 0, 7, and 28 after birth, to produce a set of reference ranges and to examine changes in these indices over the first month of life. The following dimensions were measured: interventricular septum, left ventricular posterior wall, left interventricular diameter at end systole and diastole, left atrium, and aortic root; Doppler measurements were made of maximum blood flow velocity (Vmax) through the pulmonary, aortic, mitral, and tricuspid valves. RESULTS: Reference ranges are given. Cardiac dimensions correlated well with gestation and birth weight but Vmax did not. There was a significant increase in measurements over time. The "normal" preterm infant also appeared to often have asymmetrical septal hypertrophy. Antenatal dexamethasone administration did not appear to affect the measurements. CONCLUSIONS: There is a close correlation with both gestation and birth weight for all physical measurements. Echocardiograms in preterm babies clearly differ from those in older children and adults. PMID- 9875090 TI - Severe brain injury after cardiac surgery in children: consequences for the family and the need for assistance. AB - OBJECTIVE: To identify the short and longer term needs of parents whose children sustain severe brain injury after cardiac surgery and to determine what further measures could be of use to the family after such a catastrophe. DESIGN: Qualitative analysis of data generated by semistructured interviews and a series of self report questionnaires. SETTING: Tertiary cardiothoracic referral centre. SUBJECTS: Group 1: four sets of parents (eight individuals) whose children had suffered severe brain injury after heart surgery; group 2: four sets of parents (seven individuals) caring for children with acute brain injury from other causes. RESULTS: The data provide evidence of social, emotional, physical, practical, and financial difficulties. After the children suffered brain injury following cardiac surgery their parents did not receive information, support, and practical assistance as early as they needed it. CONCLUSIONS: Although a small population was studied, it would seem that a structured, planned health care service response to this devastating event is not established; therefore, the needs of these parents are not well met. The parents and our inquiries suggest that a coordinator who is not attached to the hospital where brain injury occurred might optimally fulfil this role. PMID- 9875091 TI - A follow up study of myocardial involvement in patients with mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS). AB - OBJECTIVE: To investigate cardiac function in patients with mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) and clarify the clinical features of cardiomyopathy in MELAS. PATIENTS: 11 consecutive patients with MELAS (mean age at initial examination 11.3 years, range 4 to 16 years) were enrolled in the study. Six were followed for more than five years. RESULTS: On echocardiographic examination, three patients showed increased left ventricular end diastolic posterior wall thickness (LVPWTd), exceeding 140% of the normal value. Four patients, including these three, had an ejection fraction of less than 50%, and two also had increased left ventricular end diastolic volume (LVEDV) exceeding 140% of the normal value (%N). The LVPWTd%N was correlated positively with the LVEDV%N (R = 0.669, p < 0.05) and negatively with the ejection fraction (R = -0.6701, p < 0.05). One patient died of heart failure aged 22 years. CONCLUSIONS: The cardiomyopathy in MELAS is characterised by an abnormally thick left ventricular wall with progressive dilatation and poor left ventricular contraction developing over several years, indicating hypertrophic cardiomyopathy advancing to dilated cardiomyopathy. PMID- 9875092 TI - Nuclear cardiology in the UK 1994: activity relative to Europe, USA, and British Cardiac Society targets. AB - OBJECTIVE: To survey practice in nuclear cardiology in the UK in 1994. DESIGN: A questionnaire was sent to 219 centres performing nuclear imaging asking for details of current practice in nuclear cardiology. Replies were received from 192 centres (88%). MAIN OUTCOME MEASURES: Activity in performance of myocardial perfusion imaging (MPI) and radionuclide ventriculography (RNV), anticipated changes in activity, differences between regional and district hospitals, technical imaging parameters, and referral sources. RESULTS: Of the responding centres, 125 (65%) performed nuclear cardiology procedures. More regional than district hospitals performed nuclear cardiology procedures (85% v 55%, p < 0.0003) and regional centres performed a higher proportion (62% v 24%, p < 0.001) of nuclear cardiology activity. Nuclear cardiology activity was 0.82 scans per 1000 population per year (MPI 0.56, RNV 0.26). There has been a significant increase (24%) in nuclear cardiology since 1988. There has been a pronounced rise in MPI (350%) while RNV has fallen by 47%. Myocardial perfusion activity in the UK remains very low (25% and 5% in regional and district hospitals, respectively) compared with the 1994 figures of 2.2/1000/year for Europe or 10.8/1000/year for the USA. CONCLUSIONS: MPI has increased on average by 23%/annum (compound rate) since 1988, but in 1994 was still only 32% of the British Cardiac Society target of 2.6/1000/year. Proper resources for capital expenditure on new equipment and new staff will be important to maintain momentum in closing the gap. Also important is improved clinical understanding, as already implemented by including nuclear cardiology in guidelines for specialist cardiology training. PMID- 9875093 TI - Images in cardiology. A calcified occlusion of the left main coronary artery in a young man. PMID- 9875094 TI - Transfusion associated graft versus host disease in an immunocompetent individual following coronary artery bypass grafting. AB - Transfusion associated graft versus host disease (TA-GVHD) is a rare but commonly fatal complication of transfusion of cellular blood products, which usually occurs in immunosuppressed individuals following transfusion and subsequent engraftment of viable T lymphocytes. Very rarely it may arise in apparently immuno-competent individuals. A case is reported of TA-GVHD in a non immunocompromised 60 year old white man, resulting from red cell transfusion after coronary artery bypass grafting. HLA typing confirmed homozygosity of the donor for an HLA type shared by the recipient--the classic scenario for the development of TA-GVHD in immunocompetent individuals. The patient died 21 days after transfusion. There is a perceived increased risk of TA-GVHD following bypass grafting and other surgical procedures where cardiopulmonary bypass is required. TA-GVHD is probably underreported and the incidence in the UK is felt to be too low to warrant routine irradiation of cellular products for this group of patients. Clinicians, pathologists, and transfusion centres should be aware of this rare but devastating complication of blood transfusion after cardiac surgery. PMID- 9875096 TI - T wave alternans after sotalol: evidence for increased sensitivity to sotalol after conversion from atrial fibrillation to sinus rhythm. AB - A 64 year old woman with an 11 year history of paroxysmal atrial fibrillation presented to the emergency room because of palpitations that had started two weeks previously. She had used sotalol 80 mg once daily for three years without any episodes of proarrhythmia or other adverse effects. However, she developed pronounced T wave alternans with giant inverted T waves and excessive QT prolongation following sotalol administration one day after conversion from atrial fibrillation to sinus rhythm. This case demonstrates bizarre T wave changes, T wave alternans, and extreme QT prolongation following sotalol administration shortly after conversion from atrial fibrillation to sinus rhythm. In this situation, sotalol administration may be proarrhythmic, because it enhances repolarisation inhomogeneities based on a spatially inhomogeneous distribution of repolarisation controlling ion channels to induce repolarisation abnormalities that may lead to torsade de pointes. PMID- 9875095 TI - Successful radiofrequency catheter ablation of atrial trigeminy in a young patient. AB - A case is reported of a 35 year old man with atrial parasystolic trigeminy. The patient presented with a 10 year history of sustained supraventricular extrasystole causing symptoms leading to several hospitalisations and continuous unemployment. He had been treated ineffectively with several drug combinations. Radiofrequency catheter ablation of a right atrial focus completely suppressed the ectopic activity. This is the first report to demonstrate the efficacy and safety of radiofrequency catheter ablation in atrial ectopic trigeminy. PMID- 9875097 TI - Myocardial ischaemia in a case of a solitary coronary ostium in the right aortic sinus with retroaortic course of the left coronary artery: documentation of the underlying pathophysiological mechanisms of ischaemia by intracoronary Doppler and pressure measurements. AB - Only a few cases of a single coronary ostium and retroaortic course of the coronary artery have been described. Almost all cases reported so far had additional coronary artery or valvar disease. However, myocardial ischaemia may be caused by the coronary malformation alone. A 40 year old woman with severe myocardial ischaemia in the absence of clinically relevant coronary atherosclerosis is described. To clarify the origin and mechanisms of ischaemia, intracoronary Doppler, pressure and ultrasound studies were performed using microtransducers. In its outer portion along the course behind the ascending aorta, coronary blood flow velocities were increased, there was an external elliptical compression, and distal coronary flow reserve was reduced. Furthermore, an overshoot in diastolic pressure above aortic pressure was detectable within this portion. Dobutamine stimulation exaggerated the observed intracoronary haemodynamics and induced myocardial ischaemia. The intracoronary diagnostic procedures performed were helpful in clarifying the pathophysiological mechanisms of functional coronary obstruction and ischaemia in this malformation. Bypass surgery was successfully performed with symptomatic improvement. PMID- 9875098 TI - Cardiac valve invasion in chronic adult T cell leukaemia. PMID- 9875099 TI - Gene therapy made difficult. PMID- 9875100 TI - AAI pacing for sick sinus syndrome: first choice on all counts. PMID- 9875101 TI - In search of a reliable marker of tissue injury during heart surgery. PMID- 9875102 TI - Stethoscopes. PMID- 9875103 TI - Activated protein C resistance and myocardial infarction. PMID- 9875105 TI - Spring coil retraction in coil occlusion of persistent ductus arteriosus. AB - AIMS: To present the short and intermediate term results of coil occlusion of persistent ductus arteriosus and the results of radiographic measurements of spring coils implanted to treat patent ducts. PATIENTS: 22 children underwent coil occlusion. Their ages ranged from 2 years 9 months to 12 years 10 months (mean (SD) age, 6.5 (3.6) years). The duct diameter ranged from 1.0 to 3.5 mm at the narrowest point (mean 2.6 (0.7) mm). In 11 of the children regular coils were implanted using the non-attached system, while in the other 11 the detachable coil embolisation system was used. RESULTS: 12 children (55%) had no significant residual leaks immediately after procedures involving a single coil delivery. The remaining 10 (45%) had residual leaks immediately after the procedure, although no patient with a large duct showed residual leakage 18 months after the procedure. Radiographic measurement of the coils showed that all implanted coils retracted to 65-85% of their original size immediately after occlusion. This retraction was more evident in patients showing spontaneous closure of the residual shunt or having a coil 8 mm in diameter. CONCLUSIONS: Coil embolisation is an acceptable method for occluding persistent ductus arteriosus. Retraction of implanted coils is common in the follow up period. Such retraction may be related to spontaneous closure of residual shunt after embolisation. PMID- 9875104 TI - Myocardial perfusion defects and associated systemic ventricular dysfunction in congenitally corrected transposition of the great arteries. AB - BACKGROUND: Patients with systemic ventricles of right ventricular morphology are at high risk of contractile dysfunction, the cause of which has not been fully elucidated. OBJECTIVE: To assess whether ischaemia or infarction contributes to ventricular impairment in unoperated patients with uncomplicated congenitally corrected transposition of the great arteries (TGA) by studying myocardial perfusion and function. SETTING: Paediatric and adult congenital cardiac clinics of a tertiary referral centre. PATIENTS: Five patients with congenitally corrected TGA but without associated structural cardiac defects (aged 3.5 to 34 years). INTERVENTIONS: Maximal exercise stress testing using standard or modified Bruce protocols. Sestamibi (technetium-99m methoxy isobutyl isonitrile) scanning after isotope injection at maximal exercise and rest. MAIN OUTCOME MEASURES: Maximum exercise capacity; right ventricular myocardial perfusion, regional wall motion, and thickening; right ventricular ejection fraction. RESULTS: The two youngest patients (3.5 and 11 years) had normal exercise capacity for age, while the others had reduced exercise performance. Sestamibi scanning showed reversible myocardial ischaemia in four patients and fixed defects indicating infarction in five. Irreversible defects were mostly associated with impaired wall motion and thickening. The ejection fraction was normal (65%) in the youngest patient but < 55% in the others (mean (SD) 47(11)%). CONCLUSIONS: Patients with unoperated congenitally corrected TGA have a high prevalence of myocardial perfusion defects, with consequent abnormalities of regional wall motion and thickening, and impaired ventricular contractility. These data suggest that ischaemia and infarction are important in the pathogenesis of ventricular failure in this condition. PMID- 9875106 TI - Adenosine induced transient cardiac standstill in catheter interventional procedures for congenital heart disease. AB - OBJECTIVE: To describe the use of intravenous adenosine to create transient cardiac standstill during balloon dilatation procedures for congenital heart defects. SETTING: A tertiary paediatric cardiac centre. DESIGN AND PATIENTS: This was a prospective pilot study. Thirteen patients born with congenital heart disease and who had stenotic lesions requiring relief were considered for the technique. All were suitable for balloon dilatation. Their ages ranged from 2 months to 30 years, mean (SD) 9.9 (9.8) years. The dose of adenosine varied from 0.125 mg/kg to 0.555 mg/kg, mean 0.33 (0.127). RESULTS: Two patients only developed sinus bradycardia in response to adenosine, which may have been related to the technique of administration. The other 11 experienced a period of asystole, which ranged from 2.4 to 10.8 seconds, mean 4.99 (2.27), and a total atrioventricular block period of 5.0 to 21.2 seconds, mean 9.47 (4.64). The interval between adenosine injection and the onset of asystole varied from 2.4 to 15.8 seconds, mean 8.05 (3.6), depending on cannula size, site of administration, and cardiac output. The peak gradient across the stenotic lesions fell from 52.3 (23.7) to 17.8 (11.9) mm Hg (p < 0.001). Apart from one short episode of atrial fibrillation there were no complications. CONCLUSIONS: Intravenous adenosine is a safe and effective agent for creating transient cardiac standstill during balloon dilatation procedures for congenital heart disease. This achieves stability which is likely to improve results and reduce complications. It may have applications in other fields of cardiac intervention where an immobile heart is desirable during the critical phase of a procedure. PMID- 9875107 TI - von Willebrand factor, tissue plasminogen activator, and dehydroepiandrosterone sulphate predict cardiovascular death in a 10 year follow up of survivors of acute myocardial infarction. AB - BACKGROUND: Haemostasis plays a major part in the process initiating a myocardial infarction. The impact of haemostatic variables on long term prognosis is unknown. OBJECTIVE: To evaluate von Willebrand factor (vWF), tissue plasminogen activator antigen (t-PA) and its activity before and after venous occlusion, plasminogen activator inhibitor (PAI-1), dehydroepian-drosterone sulphate (DHEAS), and established clinical risk factors as long term predictors for reinfarction and mortality. PATIENTS: 123 consecutive survivors of myocardial infarction followed up for 10 years. DESIGN: Study entry took place between 1982 and 1983. Fifty seven patients died (54 of cardiovascular disease) during the mean observation time of 10 years. RESULTS: Cox's univariate regression analysis showed that cardiovascular mortality was significantly associated with age, hypertension, previous history of angina pectoris, DHEAS, mass concentration of t PA, and vWF. These associations were significant for vWF and mass concentration of t-PA after adjusting for age and hypertension. CONCLUSIONS: A low concentration of DHEAS and high levels of the endothelially derived haemostatic variables vWF and mass concentration of t-PA are predictors of cardiovascular mortality in survivors of myocardial infarction. This association is independent of established clinical risk factors for mass concentration of t-PA and vWF. PMID- 9875108 TI - Prevalence of factor V Leiden (APCR) and other inherited thrombophilias in young patients with myocardial infarction and normal coronary arteries. AB - OBJECTIVE: To investigate the role of activated protein C resistance (APCR, factor V Leiden) in coronary artery thrombosis. METHODS: The prevalence of APCR and of congenital deficiencies of antithrombin III, protein C, protein S, plasminogen, and factor XII was investigated in adult patients under 45 years of age with acute myocardial infarction. The results were compared with those of a group of 53 age and sex matched control subjects. RESULTS: Among 75 patients under the age of 45 years who were admitted from November 1994 to April 1996 for acute myocardial infarction, 22 (29.3%) had normal coronary arteriography (group I) and 53 (70.7%) had significant coronary artery disease (group II). Inherited thrombophilia was more often found in group I (4/22, 18.2%) than in group II (4/53, 7.5%) but the difference was not significant (F test: p = 0.22). The prevalence of APCR was 9.1% (2/22) in group I, 3.8% (2/53) in group 2 (p = 0.57), and 3.8% (2/53) in the normal control group (p = 0.57). CONCLUSIONS: The prevalence of congenital thrombophilias, including APCR, does not seem to be increased in young patients with myocardial infarction and normal coronary angiograms, compared with young patients with coronary atherosclerosis and with normal control subjects. However, the statistical power of the study is too low to detect a significant difference and these results are published to allow a meta-analysis of this problem in the future. PMID- 9875109 TI - Intraoperative release of troponin T in coronary venous and arterial blood and its relation to recovery of left ventricular function and oxidative metabolism following coronary artery surgery. AB - OBJECTIVE: To investigate the intraoperative release of troponin T during uncomplicated coronary artery surgery and to determine its relation to ischaemic time and to recovery of left ventricular function and oxidative metabolism. DESIGN: A prospective observational study. SETTING: Cardiac surgical unit in a tertiary referral centre. METHODS: Troponin T, creatine kinase, and lactate were analysed from arterial and coronary sinus samples taken before operation, and 1, 4, 6, 10, 20, 35, and 45 minutes after cross clamp release. Net myocardial troponin T release and lactate extraction were derived from their respective arteriovenous differences. Haemodynamic measurements were made using a thermodilution pulmonary artery catheter. PATIENTS: 45 patients, mean (SD) age 62 (9) years, with two or three vessel coronary artery disease and chronic stable angina undergoing routine coronary artery surgery. RESULTS: Before operation, troponin T concentrations were not raised, but within one minute of cross clamp release they increased progressively in both coronary sinus and arterial blood for the entire 45 minutes of reperfusion studied. Coronary sinus troponin T concentrations were consistently higher than arterial concentrations at all time points (p < 0.001), indicating net troponin T release by the myocardium. Peak net troponin T release and area under the curve of net troponin T release correlated closely with ischaemic time (r = 0.58 and r = 0.61, p < 0.0001 for both). Area under the curve of arterial troponin T concentration was also significantly correlated with ischaemic time (r = 0.44, p < 0.01). Patients with cross clamp times longer than 72 minutes (upper quartile for ischaemic time) had greater troponin T release, delayed reversion to lactate extraction, and lower left ventricular stroke work index three hours after surgery, compared with patients who had short (< 50 minutes, lower quartile) and intermediate (51-71 minutes, interquartile) cross clamp times. Peak net troponin T release and area under the curve of arterial troponin T concentration were inversely correlated with left ventricular stroke work index three hours after surgery (r = -0.57, r = -0.38, p < 0.01). CONCLUSIONS: Troponin T concentrations increased in every patient after cross clamp release, and were consistently higher in coronary sinus blood than in arterial blood, indicating net myocardial release of troponin T during the period of reperfusion. Intraoperative net troponin T release has functional significance, as it is closely related to ischaemic time and reflects delayed recovery of left ventricular function and oxidative metabolism; therefore, its measurement may contribute to the perioperative assessment of myocardial injury sustained during coronary artery surgery. PMID- 9875110 TI - Comparison of peripheral endothelial dysfunction and intimal media thickness in patients with suspected coronary artery disease. AB - OBJECTIVE: Flow associated dilatation (FAD%) and intimal media thickness are established markers of early atherosclerosis. This study aimed to compare the ability of the non-invasive measurements FAD% and intimal media thickness to predict coronary artery disease. METHODS: FAD% and intimal media thickness were determined using high resolution ultrasound in 122 patients with clinically suspected coronary artery disease before coronary angiography. Results are given as mean (SD). RESULTS: Patients with coronary artery disease had reduced FAD% compared with those with angiographically normal coronary vessels (3.7 (4.1) v 7.0 (3.5)%, p < 0.001), whereas intimal media thickness tended to be increased in patients with coronary artery disease (0.58 (0.35) v 0.47 (0.11)mm, p = 0.054). There was a negative correlation between FAD% and intimal media thickness (R = 0.317, p = 0.0004). Receiver operating characteristic analysis showed that FAD% < or = 4.5% predicted coronary artery disease with a sensitivity of 0.71 (95% confidence interval 0.61 to 0.80) and a specificity of 0.81 (0.58 to 0.95). In contrast, intimal media thickness showed a positive correlation with the extent of coronary artery disease (number of vessels with a lesion > or = 50%) (R = 0.324, p = 0.0003), without a clear cut off point. CONCLUSIONS: In patients with clinically suspected coronary artery disease, FAD% discriminates between the presence or absence of coronary artery disease, whereas intimal media thickness is associated more with the extent of coronary artery disease. PMID- 9875111 TI - Endoluminal vessel reconstruction by stent placement in patients with obstructive coronary artery disease who are poor surgical candidates. AB - OBJECTIVE: To investigate the safety and efficacy of endoluminal vessel reconstruction by intracoronary stenting in patients with single or multivessel disease who have been turned down for surgical revascularisation. DESIGN: An observational, single centre trial of consecutive patients being followed up clinically. SETTING: Tertiary referral centre. PATIENTS: 43 patients treated between 1995 and 1996 with a clinical follow up period of at least six months. MAIN OUTCOME MEASURES: Acute procedural success following intervention (defined as a residual stenosis of < 30% with normal antegrade flow and without acute clinical complications); major adverse clinical events (death, myocardial infarction, repeat intervention, clinical restenosis); and functional status during follow up. RESULTS: Acute procedural success was achieved in 39 patients. Failures resulted from a refractory no reflow phenomenon with Q wave myocardial infarction (two patients), and from subacute stent thrombosis (one patient) and saphenous vein graft rupture at stent site (one patient), both manifesting by non Q wave myocardial infarction. During a follow up period of 17 (8) months (mean (SD)), three of these patients died. Clinical restenosis was treated by repeat percutaneous intervention in 11 patients, while 28 patients remained asymptomatic. On a Kaplan-Meier estimate, the proportion of patients without adverse clinical events at two years was 65%. CONCLUSIONS: Percutaneous reconstruction of diseased coronary arteries in patients with single or multivessel disease turned down for surgical revascularisation may be considered as a valuable treatment option. On follow up, more events seem to be encountered than after elective de novo stenting of discrete coronary lesions. This is probably related to higher clinical restenosis rates. PMID- 9875113 TI - Nitrate induced coronary vasodilatation: differential effects of sublingual application by capsule or spray. AB - BACKGROUND: Sublingual nitroglycerin (glyceryltrinitrate, GTN) capsules or isosorbide dinitrate (ISDN) spray are routinely used to treat anginal attacks and to vasodilate maximally the epicardial coronary arteries during coronary angiography. OBJECTIVE: To compare the coronary vasodilatory effects of GTN capsules and ISDN spray with those induced by intracoronary GTN using quantitative coronary angiography. DESIGN: 96 patients (79 men and 17 women; median age 59 years) were randomised to four groups to receive either a sublingual capsule containing 0.8 mg GTN or two puffs of spray delivering 0.8 mg ISDN, followed or preceded by an intracoronary bolus of 0.2 mg GTN used as reference for maximal vasodilatation. RESULTS: There was a significant increase in the mean diameter of coronary arteries in angiographically normal segments in patients who received either intracoronary GTN (groups 1 and 2) or ISDN spray (group 4) as a first application (group 1, 0.46 mm, + 17%, (baseline vessel diameter 100%), p < 0.001; group 2, 0.45 mm, + 13%, p < 0.001; group 4, 0.47 mm, + 13%, p < 0.05). Patients who received a sublingual GTN capsule as the first application mode (group 3) had no significant change in epicardial vessel diameter (0.10 mm, + 5%, p = 0.3). CONCLUSIONS: Sublingual ISDN spray may be more efficacious than sublingual GTN capsules in certain patients with anginal attacks. ISDN spray should be preferred over capsules in coronary angiographic procedures. PMID- 9875112 TI - Predictors of event-free survival after percutaneous mitral commissurotomy. AB - OBJECTIVE: To assess the long term functional result after percutaneous mitral commissurotomy and identify the predictors of event-free survival following 10 years of experience. DESIGN: Analysis of clinical, echocardiographic, and haemodynamic variables at baseline and after the procedure by univariate and multivariate analyses (Cox model). SETTING: University hospital. PATIENTS: 532 consecutive patients receiving percutaneous mitral commissurotomy in the same institution. RESULTS: The mean (SD) follow up was 3.8 (4.0) years. Survival at 3, 5, and 7.5 years was 94%, 91%, and 83%, respectively; event-free survival was 84%, 74%, and 52%. Mitral valve anatomy was identified as the strongest independent predictor of event-free survival. Age, cardiothoracic ratio, mean pulmonary artery pressure, and mean echocardiographic mitral gradient after commissurotomy were also found to be independent predictors of long term functional results. Event-free survival was 92%, 84%, and 70% at 3, 5, and 7.5 years in patients with favourable anatomy (echo score = 1), 86%, 73%, and 34% in patients with intermediate anatomy (echo score = 2), and 45%, 25%, and 16% in patients with unfavourable anatomy (echo score = 3). In patients aged < or = 65 years, the event-free survival rate was 80%, 70%, and 45% at 3, 5, and 7.5 years v 52%, 38%, and 17% in patients aged > 65 years. CONCLUSIONS: The anatomical form of the mitral valve and the patient's age were the most powerful predictors of event-free survival. Patients with intermediate or unfavourable anatomy and those aged > 65 years have low 5 and 7.5 year event-free survival rates. This must be taken into account when discussing the indications for percutaneous mitral commissurotomy; immediate mitral valve replacement is a reasonable alternative to balloon mitral commissurotomy in patients with higher risk of functional deterioration after the procedure. PMID- 9875114 TI - Images in cardiology. Fibrosarcoma of the heart. PMID- 9875116 TI - Images in cardiology. Idiopathic chylopericardium. PMID- 9875115 TI - Selection of the optimal stress test for the diagnosis of coronary artery disease. AB - OBJECTIVE: To compare the value and limitations of exercise testing, dipyridamole echocardiography, dobutamine-atropine echocardiography, and MIBI-SPECT (technetium-99m methoxyisobutyl nitrile single photon emission computed tomography) during dobutamine infusion in the diagnosis of coronary artery disease. DESIGN: The performance of these four tests was assessed in random order on a consecutive cohort of patients. The presence or absence of coronary artery disease was confirmed by coronary angiography. SETTING: Two tertiary care and university centres. PATIENTS: 102 consecutive patients with chest pain and no previous history of coronary artery disease. Ten patients with left bundle branch block were excluded for further analysis of exercise testing and scintigraphy results. RESULTS: MIBI-SPECT was the most sensitive (87%) but the least specific test (70%). Exercise stress testing had a sensitivity of 66%, which increased to 80% when patients with inconclusive results were excluded. Dipyridamole and dobutamine echocardiography had similar sensitivity (81%, 78%) and specificity (94%, 88%). All four tests had similar accuracy and positive and negative predictive values. Agreement between the echocardiographic techniques was excellent (detection of coronary artery disease 87%, kappa = 0.72; regional analysis 93%, kappa = 0.72; diagnosis of the "culprit" vessel 95%, kappa = 0.92), and it was good between echocardiographic techniques and MIBI-SPECT (diagnosis of the culprit vessel 90%, kappa = 0.84 with dobutamine and 92%, kappa = 0.85 with dipyridamole). CONCLUSIONS: Exercise stress testing has a sensitivity comparable to other tests in patients capable of exercising and with no basal electrical abnormalities. The greatest sensitivity is offered by MIBI-SPECT and the greatest specificity is obtained with stress echocardiography. Redundant information is obtained with dipyridamole echocardiography, dobutamine echocardiography, and MIBI-SPECT. PMID- 9875117 TI - Reproducibility and responsiveness of quality of life assessment and six minute walk test in elderly heart failure patients. AB - OBJECTIVE: To examine the reproducibility and responsiveness to change of a six minute walk test and a quality of life measure in elderly patients with heart failure. DESIGN: Longitudinal within patient study. SUBJECTS: 60 patients with heart failure (mean age 82 years) attending a geriatric outpatient clinic, 45 of whom underwent a repeat assessment three to eight weeks later. MAIN OUTCOME MEASURES: Subjects underwent a standardised six minute walk test and completed the chronic heart failure questionnaire (CHQ), a heart failure specific quality of life questionnaire. Intraclass correlation coefficients (ICC) were calculated using a random effects one way analysis of variance as a measure of reproducibility. Guyatt's responsiveness coefficient and effect sizes were calculated as measures of responsiveness to change. RESULTS: 24 patients reported no major change in cardiac status, while seven had deteriorated and 14 had improved between the two clinic visits. Reproducibility was satisfactory (ICC > 0.75) for the six minute walk test, for the total CHQ score, and for the dyspnoea, fatigue, and emotion domains of the CHQ. Effect sizes for all measures were large (> 0.8), and responsiveness coefficients were very satisfactory (> 0.7). Effect sizes for detecting deterioration were greater than those for detecting improvement. CONCLUSIONS: Quality of life assessment and a six minute walk test are reproducible and responsive measures of cardiac status in frail, very elderly patients with heart failure. PMID- 9875118 TI - "Value" of improved treadmill exercise capacity: lessons from a study of rate responsive pacing. AB - OBJECTIVES: To compare the value of a series of cardiovascular measurements in patients with symptomatic disease receiving an effective treatment (rate responsive pacing). PATIENTS: 12 pacemaker dependent patients with VVIR units. INTERVENTIONS: Single blind crossover between VVI and VVIR. OUTCOME MEASURES: Exercise capacity was assessed by treadmill tests (modified Bruce protocol and a fixed workload protocol) with respiratory gas analysis. Self paced corridor walk tests were also undertaken. Quality of life (QOL) was assessed by questionnaire. Daily activity was measured in the patients' homes using shoe and belt pedometers. RESULTS: Treadmill tests and QOL questionnaires correctly identified the clinical benefit associated with VVIR. The modified Bruce protocol was superior to the fixed workload protocol as it was better tailored to the fairly well preserved exercise capacity of the patients. Symptom scores, but not walking times, were improved with VVIR during corridor walk tests. VVIR did not improve daily activity measured using either the belt or shoe pedometers. CONCLUSIONS: VVIR pacing improved some but not all measures of exercise capacity. This finding illustrates the difficulty of selecting an instrument to measure symptomatic improvement in clinical research; and raises the question, what is the best way of measuring exercise capacity? PMID- 9875119 TI - Single chamber atrial pacing: an underused and cost-effective pacing modality in sinus node disease. AB - OBJECTIVE: To determine the safety and cost effectiveness of single chamber atrial pacing in patients with sinus node disease. DESIGN: Retrospective follow up study. SETTING: Tertiary referral centre. PATIENTS: 81 patients with single chamber atrial pacemakers implanted between 1992 and 1996. MAIN OUTCOME MEASURES: The development of high grade atrioventricular block resulting in a further pacemaker procedure. The cost savings of changing our current pacing practice to conform with British Pacing and Electrophysiology Group guidelines. RESULTS: During the follow up period, four patients (5.8%) required a further procedure to upgrade their atrial pacemaker to a dual chamber system owing to the development of high grade atrioventricular block. In 1995 and 1996, 343 pacemakers were implanted in patients with sinus node disease; 19 (5.5%) received single chamber atrial pacemakers and 271 (79%) dual chamber pacemakers. If the current pacing practice was changed so that all patients received single chamber atrial pacemakers, with revision for symptomatic atrioventricular block, savings in excess of 206,000 Pounds would have been made in the two year period. CONCLUSIONS: Atrial pacing in patients with sinus node disease is underused. The need for patients to undergo further procedures owing to the development of atrioventricular block is small and significant cost savings could be made by changing pacemaker practice. PMID- 9875120 TI - Predictors of loss of atrioventricular synchrony in single lead VDD pacing. AB - OBJECTIVE: To evaluate maintenance of proper VDD function, defined as persistence of sinus rhythm with atrial synchronous ventricular pacing, and to define factors predicting failure of the VDD mode in patients with atrioventricular (AV) block and normal sinus function. DESIGN: Observational study in 86 consecutive patients (mean (SD) age 74 (12) years; 38 women, 48 men) with single lead VDD pacing systems (Intermedics Unity, n = 66, Medtronic Thera VDD, n = 20), implanted for high degree AV block with documented normal sinus node. Pacemaker function was assessed by event counters, telemetric measurements, and Holter recordings. Demographic, radiological, and pacing variables were correlated with loss of proper VDD function. RESULTS: During a mean (SD) follow up of 10 (10) months (range 1-37), sinus rhythm and atrial triggered ventricular pacing were maintained in 70 of 86 patients (81%). Atrial undersensing was observed in nine patients, lead migration in two, atrial fibrillation in three, and symptomatic sinus bradycardia in two. Univariate predictors of loss of proper VDD function were: low position of the atrial dipole relative to the carina (> or = 6 cm; p < 0.01) during fluoroscopy; and maximum programmable atrial sensitivity of the pacemaker (p = 0.03). In a multivariate analysis, only dipole position remained predictive of outcome (p < 0.02). Not predictive were sex, age, symptoms before pacemaker implantation, cardiothoracic ratio or dilatation of individual heart chambers on chest x ray, side of device implant, and P wave amplitude at implant. CONCLUSIONS: To maintain proper VDD function in the long term, a low anatomical dipole position relative to the carina should be avoided. Electrical guidance of dipole positioning does not seem to influence long term outcome. PMID- 9875121 TI - Antiplatelet treatment with cilostazol after stent implantation. AB - OBJECTIVES: To evaluate the efficacy of cilostazol, a new synthetic inhibitor of phosphodiesterase, in preventing stent thrombosis after successful implantation. DESIGN: Preliminary prospective study. SETTING: A single coronary care unit in Japan. PATIENTS: Elective, bailout, or primary stents were implanted in 85 consecutive patients with 93 lesions. Primary stent implantation was performed in 18 patients with acute myocardial infarction. Patients received 200 mg cilostazol and 243 mg aspirin after stenting. MAIN OUTCOME MEASURES: Stent thrombosis, major and minor complications, and side effects were assessed in the six months after stenting. RESULTS: Gianturco-Roubin stents were implanted in 37 lesions, Wiktor stents in 55, and Palmaz-Schatz stents in 27. Multiple stents were used in 26 lesions. There was no mortality, stent thrombosis related Q wave myocardial infarction, emergency bypass surgery, repeat intervention, or vascular complications in the six months of follow up. Acute or subacute closure did not occur after stenting. There were no serious side effects such as leucopenia and/or abnormal liver function for three months. Cilostazol was withdrawn in one patient because of skin rash. Patients who underwent primary stenting had no clinical events, such as acute or subacute thrombosis, or side effects. CONCLUSIONS: Cilostazol is an effective antiplatelet agent with minimum side effects after elective, bailout, or primary stent implantation. PMID- 9875122 TI - Approaches to modern management of cardiac arrest. PMID- 9875123 TI - Public access defibrillation: a review. PMID- 9875124 TI - Determinants of successful defibrillation. PMID- 9875125 TI - Antiarrhythmic drugs in resuscitation. PMID- 9875126 TI - Adrenaline dosage and buffers in cardiac arrest. PMID- 9875127 TI - Raised factor VIII is associated with coronary thrombotic events. AB - Coagulation is triggered during the onset of myocardial infarction, resulting in vascular occlusion. However, a causal role for individual haemostatic factors in the development of thrombotic occlusion is not established. Three cases (all relatively young women) are reported of raised factor VIII associated with myocardial infarction. Two patients presented acutely with myocardial infarction at a relatively young age with no preceding history of angina. The other patient had had venous thrombosis when young and activated protein C resistance (APCR), without the presence of factor V Leiden. A functional relation exists between APCR and factor VIII; therefore, raised factor VIII may contribute to APCR and the increased thrombotic risk in patients without factor V Leiden. Factor VIII is an important risk factor for atherothrombotic events, including sudden death, in patients with vascular disease. These cases support the association of raised factor VIII with acute thrombotic events, even in patients without significant underlying atheromatous disease. PMID- 9875128 TI - Intermittent complete atrioventricular block associated with typical atrioventricular nodal reentrant tachycardia. PMID- 9875129 TI - Images in cardiology. Coronary artery fistula. PMID- 9875130 TI - Subpectoral implantation of a cardioverter defibrillator under local anaesthesia. PMID- 9875131 TI - Ventricular pacemaker upgrade: experience, complications, and recommendations. PMID- 9875132 TI - Integrins, cadherins, and catenins: molecular cross-talk in cancer cells. AB - In the past decade, there have been major advances in the understanding of some of the mechanisms underlying tumour differentiation, invasion, and metastasis, in which cell-cell and cell-matrix adhesion molecules play a critical role. Cadherin/catenin complex and the integrins are the prime mediators of cell adhesion in normal and transformed cells, cadherin/catenin being largely responsible for intercellular adhesion and integrins for cell-extracellular matrix interactions. Intercellular and cell-matrix adhesion mediated by cadherin/catenin and integrins is likely to play a role in the control of both structural morphology and functional differentiation; hence, any loss of this control mechanism may well facilitate the neoplastic process. Indeed, in cancer cells, there is a co-ordinated down-regulation of both integrins and cadherins which correlates with tumour dedifferentiation. However, the expression and cellular localization of catenins do not always correlate with cadherin expression, since the catenins are rather promiscuous molecules which interact not only with E-cadherin, but also with growth regulatory and signalling molecules such as epidermal growth factor receptor and the adenomatous polyposis coli gene product. PMID- 9875133 TI - Clear cell sarcoma of tendons and aponeuroses (malignant melanoma of soft parts) and cutaneous melanoma: exploring the histogenetic relationship between these two clinicopathological entities. AB - Clear cell sarcoma of tendons and aponeuroses (CCS) has come to be recognized as a distinct histopathological entity in the last three to four decades. It shares a number of histological and ultrastructural features with cutaneous melanoma (MM), occasionally creating diagnostic difficulties with metastatic melanoma in the absence of a known primary cutaneous tumour. At a genetic level, a t(12;22) has been identified in 60-75 per cent of cases of CCS using karyotype analysis, while MM demonstrates a broad range of genetic alterations, most commonly appearing to involve chromosomes 1, 5, and 6. Although these two tumour types share many common microscopic, and thus histogenetic, similarities, the genotypic evidence supports two distinct histopathological entities. PMID- 9875134 TI - Comparison of integrin, cadherin, and catenin expression in squamous cell carcinomas of the oral cavity. AB - In addition to their role in maintenance of tissue integrity, cell adhesion molecules regulate the growth and differentiation of stratified squamous epithelia. Reduced expression of E-cadherin and the alpha 2 beta 1, alpha 3 beta 1 and alpha 6 beta 4 integrins is already reported to correlate with poor histological differentiation in oral squamous cell carcinomas. However, it is not clear how closely cadherin and integrin loss are related in any given tumour, nor whether cadherin loss is correlated with changes in expression of the cytoplasmic regulatory proteins known as catenins. Double-label immunofluorescence has been used to stain a panel of 22 oral squamous cell carcinomas with antibodies to ten proteins, including E- and P-cadherin, the major keratinocyte integrin subunits, and alpha-, beta- and gamma-catenin. Overall, E-cadherin expression and integrin expression correlated well with tumour grade, while P-cadherin staining was more variable. All tumours, regardless of differentiation status, showed reduced staining for at least two of the catenins, implying that the adhesive function of E- and P-cadherin could be impaired even when cadherin expression is normal. It is concluded that in all squamous cell carcinomas, regardless of degree of histological differentiation, there is some perturbed expression of cell adhesion molecules and that integrin and E-cadherin loss are closely related. PMID- 9875135 TI - Disorderly CD44 gene expression in human cancer cells can be modulated by growth conditions. AB - Disorderly CD44 gene expression is a well-documented characteristic feature of tumour cells from cancers arising in many different organs of the human body. Molecular pathological studies have established that the pattern of the abnormal expression can differ according to the origin and the stage of the tumour. In this investigation it has been demonstrated that in some but not all tumour cell lines, which are undeniably and irreversibly malignant when inoculated in vivo, CD44 gene expression can still be modulated. In two cell lines, the pattern of CD44v expression was found to be affected by cell-to-cell and cell-to-substrate attachment. Expression was up-regulated by cell-substrate interactions, but only until cell-to-cell contact caused subsequent down-regulation of CD44v transcription. This information provides new opportunities for detailed investigation of the mechanisms of abnormal CD44 gene regulation in cancer and for exploring stage-related changes in the expression of this complex gene. PMID- 9875136 TI - p53-dependent X-ray-induced modulation of cytokine mRNA levels in vivo. AB - In vitro studies have shown that ionizing radiation can cause increases in some cytokine mRNA levels and activation of the nuclear NF-kappa B and/or AP1 transcription factors which have been implicated in the transcriptional regulation of many cytokine genes. Thus, radiation-induced upregulation of cytokine mRNAs appeared to be in part a direct consequence of transcription factor activation. To test this in vitro model in vivo, the effects of whole-body X-irradiation (0-10 Gy) on cytokine and other gene mRNA levels have been examined in mice. Increases and decreases in cytokine mRNA levels were detected in tissues which underwent an early wave of apoptosis (bone marrow and/or spleen), but not in more radioresistant tissues (kidney, liver, brain, and heart). Some mouse strain-specific differences were observed, but none of the changes in mRNA level was detected in p53-/- mice. As activation of the NF-kappa B and AP1 transcription factors was not detected in early-(spleen) or late-(liver) responding tissues in 10 Gy X-irradiated p53+/+ mice in vivo, it is concluded that the modulation of cytokine gene expression in vivo is p53-dependent and indirectly associated with apoptosis. PMID- 9875137 TI - No difference in stem cell somatic mutation between the background mucosa of right- and left-sided sporadic colorectal carcinomas. AB - Epidemiological, morphological, and molecular differences exist between carcinomas of the right and left sides of the large bowel. To investigate whether this is reflected in differences in somatic mutation frequency in the background mucosa, mutation of the neutral O-acetyltransferase gene (oat) was quantified in histologically normal resection margins from 20 informative (heterozygous) patients with caecal or ascending colon cancer (11 males, median age 75 years) and 20 with sigmoid colon or rectal cancer (10 males, median age 70 years). Mutant discordant crypts lacking O-acetyltransferase activity were visualized by mPAS staining and classified as wholly or partially involved by the mutant phenotype; median frequencies (x10(-4) were compared (Mann-Whitney U-test) after assessing a sample of more than 10,000 crypts per case. No significant difference was found between the frequencies of wholly involved mPAS-positive crypts in background mucosa of left- and right-sided cancers (p = 0.4569), indicating that tumours on both sides of the colon are associated with similar levels of lifetime accumulated stem cell mutational load. However, partially involved mPAS-positive crypts were significantly more frequent in mucosa from left-sided cancers (p < 0.04), indicating increased mutational activity during the previous 12 months. Analysis of mucosa proximal and distal to left-sided cancers showed that this increase was due to a statistically higher frequency of partially involved crypts in proximal mucosa, which probably resulted from the obstructive effects of the tumour causing increased exposure of the proximal mucosa to luminal carcinogens and/or epithelial regeneration in response to low-grade inflammation or ischaemia. The findings indicate that although left-sided colonic cancer is commoner than right-sided cancer in the British population, carcinomas on both sides of the large bowel arise in a background of similar levels of stem cell mutational activity. PMID- 9875138 TI - No evidence for functional inactivation of wild-type p53 protein by MDM2 overexpression in gastric carcinogenesis. AB - Inactivation of wild-type p53 during gastric carcinogenesis is usually caused by mutations within exons 5-8 of the p53 gene leading to mutated, usually immunohistochemically detectable p53 proteins. However, functional inactivation of wild-type p53, mimicking mutational inactivation, may also result from binding to overexpressed MDM2 protein. While these two mechanisms of p53 inactivation are considered to be mutually exclusive, no data exist as to whether MDM2 overexpression occurs during gastric carcinogenesis. MDM2 protein overexpression was therefore studied in relation to p53 protein accumulation in gastric carcinogenesis. Forty-five paraffin-embedded gastrectomy specimens from early gastric carcinomas were examined for the presence of chronic active gastritis, chronic atrophic gastritis, subtypes of intestinal metaplasia, and dysplasia. The Lauren type was reassessed for all early carcinomas. p53 protein accumulation was examined using the monoclonal antibody DO-7. MDM2 protein overexpression was assessed with the monoclonal antibody SMP-14. Complete absence of nuclear p53 protein accumulation was observed in chronic active gastritis, chronic atrophic gastritis, and intestinal metaplasia, irrespective of the subtype. In gastric dysplasia (one mild, two moderate, one severe), only severe dysplasia was p53 positive. Intestinal-type (n = 20) and diffuse-type early gastric carcinoma (n = 25) were p53-positive in 70 and 52 per cent of the cases, respectively. MDM2 protein overexpression was not observed during gastric carcinogenesis, either in the p53-positive or in the p53-negative cases. In conclusion, it appears that functional inactivation of wild-type p53 by MDM2 protein overexpression plays no role in (early) gastric carcinogenesis. PMID- 9875139 TI - Pancreatic endocrine tumours: evidence for a tumour suppressor pathogenesis and for a tumour suppressor gene on chromosome 17p. AB - Two molecular pathways leading to cancer are known. Common-type cancers arise from the 'tumour suppressor' pathway, characterized by gross chromosomal changes and allelic losses (LOH) in an average of 25 per cent or more of randomly chosen chromosomal loci. The 'mutator pathway' has been recognized in a subset of cancers, characterized by widespread microsatellite DNA instability and rarity of chromosomal losses. The present study has investigated 20 pancreatic endocrine tumours (PETs) for loss of heterozygosity (LOH) at seven chromosomal loci (3p14, 7q31-32, 11q13, 13q14, 18q21, 17p13, and 17q21); microsatellite instability; and Ki-ras, N-ras, and p53 gene mutations. LOH was found in an average of 24 per cent of the chromosomal loci analysed. No tumour showed microsatellite instability. Ki ras and p53 mutations were each found in one case. The frequency of losses was higher in malignant (40 per cent) than in benign (17 per cent) tumours (p = 0.009), and the specific chromosome 17p13 LOH was associated with extrapancreatic extension of disease (p = 0.007), high proliferative activity (p = 0.001), and absence of progesterone receptors (p = 0.01). A common deleted region on chromosome 17p13 and the rarity of p53 gene mutations suggest the existence of a novel tumour suppressor gene involved in the pathogenesis of PETs in this chromosomal area. PMID- 9875140 TI - Prognostic value of MMP-2 immunoreactive protein (72 kD type IV collagenase) in primary skin melanoma. AB - The penetration of the subepithelial basement membrane is the first critical step in the dissemination of melanoma. In vitro studies have suggested that the 72 kD type IV collagenase (MMP-2) may be important in melanoma invasion. It has recently been demonstrated that the expression of MMP-2 immunoreactive protein increased with increasing atypia in melanocytic tumours and was associated with later haematogenous metastases in melanoma. This paper investigates the value of MMP-2 as a possible prognostic marker in melanoma. The expression of MMP-2 immunoreactive protein was studied with immunoperoxidase staining in paraffin embedded sections of 50 cases of primary skin melanoma by using specific, affinity purified antibodies. Positive immunostaining was quantified by counting the percentage of positive cancer cells and was compared with clinical patient characteristics and survival. Sixty-four per cent of the primary melanoma cases displayed positive cytoplasmic immunostaining for MMP-2 in tumour cells. Marked overexpression of MMP-2 protein (> or = 34 per cent of melanoma cells positive) correlated with the 5-year survival of the patients when compared with patients with lower MMP-2 positivity, 55 per cent vs. 85 per cent, respectively (P < 0.05). Male patients displayed positive staining more often than females (75 per cent vs. 54 per cent, respectively). There was no correlation between MMP-2 positivity and Clark level or Breslow classification. A distinct group with unfavourable prognosis was identified. The 10-year survival for MMP-2-positive male melanoma patients was 39 per cent as opposed to 79 per cent with the other melanoma patients (P < 0.05). In the hierarchic Cox regression model for survival, MMP-2 immunoreactive protein was found to be independent of Clark level and Breslow classification. Overexpression of MMP-2 protein indicated a 4.5-fold relative risk of dying from melanoma. It is concluded that MMP-2 immunoreactive protein in melanoma cells is an independent prognostic factor for survival. High MMP-2 expression in male melanoma patients indicates an unfavourable prognosis. PMID- 9875141 TI - Human mast cells express stem cell factor. AB - Stem cell factor (SCF) is a major cytokine regulator of mast cell growth and function. The present study demonstrates that human mast cells are able to produce SCF. Constitutive synthesis of SCF mRNA was seen in the mast cells isolated from human lung and skin by RT-PCR. This was confirmed by in situ hybridization in conjunctival mast cells of both tryptase-only (MCT) and tryptase/chymase (MCTC) subsets. SCF protein product was found in conjunctival MCT and MCTC mast cells by immunohistochemistry. Soluble SCF protein was detected in the culture supernatant of isolated lung mast cells by ELISA, and cross linkage of IgE receptor (Fc epsilon-RI) on the lung mast cells in culture did not alter SCF mRNA expression, or the secreted soluble SCF protein. This was consistent with the finding that levels of SCF mRNA expression in conjunctival mast cells were similar between normal subjects and patients with seasonal allergic conjunctivitis (SAC). This study shows that human mast cells themselves are a cellular source of SCF, as well as being target cells for this growth factor. SCF may regulate mast cell growth and function via both paracrine and autocrine mechanisms. The production of SCF by mast cells may be regulated via mechanisms other than IgE receptor-mediated pathways. PMID- 9875142 TI - Mast cell subpopulations in the synovial tissue of patients with osteoarthritis: selective increase in numbers of tryptase-positive, chymase-negative mast cells. AB - Although there is relatively little evidence of inflammation in osteoarthritis (OA), increases in mast cell numbers and mast cell activation are prominent features of the synovial tissue. As little is known of the types of mast cells which may be involved, the numbers and distribution of mast cell subpopulations have been investigated as defined according to their content of proteases. Tissue was obtained from patients with OA undergoing total knee replacement surgery (n = 14) and from control subjects either post-mortem (n = 11) or following leg amputation for peripheral vascular disease (n = 3); a double-labelling immunocytochemical procedure with monoclonal antibodies specific for tryptase and chymase was applied to identify those mast cells which contain both tryptase and chymase (MCTC) and those with tryptase but not chymase (MCT). There was considerable variation between individual tissues and between sites of tissue sampling, but cells of the MCTC subset were predominant in the synovial layer of both groups of subjects without joint disease, accounting for some 60 per cent of all mast cells present. In tissue from OA patients, however, there appeared to have been a striking shift in the relative proportions of mast cells from the MCTC to the MCT phenotype, with many more MCT cells present in the synovial tissues of OA patients (median 53 MCT/mm2) than in tissue from post-mortem (7.5 MCT/mm2, P < 0.0001) or amputation controls (12 MCT/mm2). In contrast, numbers of synovial MCTC cells in the synovium of OA patients (20 MCTC/mm2) differed little from those in either of the control groups (both 12 MCTC/mm2). In several other conditions, the MCT cells have been linked with inflammatory events, but it seems that in OA, other factors may be operating to induce a selective expansion of this subpopulation. PMID- 9875143 TI - Analysis of the cellular infiltrates and expression of cytokines in synovial tissue from patients with rheumatoid arthritis and reactive arthritis. AB - The cellular infiltrates and cytokine patterns in synovial tissue (ST) from patients with rheumatoid arthritis (RA) and reactive arthritis (ReA) were compared in order to determine the mechanisms responsible for the chronic and destructive course of RA. Since the results could be influenced by differences in disease duration, ST was studied from patients in both early and late stages of the disease. Ten patients had early RA (< 1 year), ten long-standing RA (> 1 year), six early ReA (< 1 year), and five long-standing ReA (> 1 year). Histological analysis demonstrated that the scores for infiltration by lymphocytes and plasma cells, and the scores for inflammation, were significantly higher in RA than in ReA. Immunolabelling studies showed that in particular, the scores for infiltration by CD38+ plasma cells, granzyme B+ cells, and interferon gamma (IFN gamma)+ cells were significantly higher in RA than in ReA. The results were independent of the disease duration. The increased number of lymphocytes, plasma cells, and granzyme B+ cells in rheumatoid synovial tissue supports the paradigm that RA is the result of specific immune recognition in the joint and that granzyme B+ cells play an important role in joint destruction. PMID- 9875144 TI - Distinct patterns of chemokine expression are associated with leukocyte recruitment in alcoholic hepatitis and alcoholic cirrhosis. AB - Alcoholic liver disease is associated with three histologically distinct processes: steatosis (parenchymal fat accumulation), alcoholic hepatitis (characterized by parenchymal infiltration by neutrophil polymorphs), and alcoholic cirrhosis (in which chronic inflammation and fibrosis dominate). Chemokines are cytokines that promote subset-specific leukoycte recruitment to tissues and could therefore play a crucial role in determining which leukocyte subsets are recruited to the liver in alcoholic liver disease. This paper reports that chemokine expression is increased in the liver of patients with alcoholic liver disease and, moreover, that distinct patterns of chemokine expression are associated with the different inflammatory responses to alcohol. Interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1 alpha (MIP-1 alpha), and MIP-1 beta were all detected in the parenchyma at sites of inflammation in alcoholic hepatitis, whereas in alcoholic cirrhosis, chemokines were restricted to inflammatory cells and endothelium in the fibrous septa and portal tracts. In alcoholic hepatitis, chemokine transcription was localized to sinusoidal cells, leukocytes, and fibroblasts in areas of parenchymal inflammation, but hepatocytes, despite staining strongly for chemokine protein, were negative. In alcoholic cirrhosis, chemokine mRNA was detected in portal tract endothelium, leukocytes, and fibroblasts. Thus, alcoholic hepatitis and alcoholic cirrhosis are associated with distinct patterns of chemokine expression that are likely to be important factors in determining whether a patient develops acute parenchymal inflammation and alcoholic hepatitis, or chronic septal inflammation and alcoholic cirrhosis. PMID- 9875145 TI - Double intratracheal instillation of keratinocyte growth factor prevents bleomycin-induced lung fibrosis in rats. AB - Alveolar re-epithelialization is necessary in the repair of damaged alveolar epithelium after lung injury. Keratinocyte growth factor (KGF) has been shown to be a potent proliferation and differentiation factor for rat alveolar type II cells. The present study examined whether KGF would prevent bleomycin-induced lung fibrosis. Adult rats were anaesthetized and recombinant human KGF (rhKGF) (150 micrograms/kg) or saline was injected intratracheally at 48 h before and 24 h after bleomycin (Bleo, 5 mg/kg) instillation. Seven and 14 days after the last administration, rat lungs were processed for lung physiology, immunohistochemistry, and in situ hybridization. Double instillation of KGF prevented the loss of body weight and reduction in total lung capacity (TLC) due to Bleo, and markedly attenuated the protein accumulation and mRNA expression of collagen types I and III and the decreased expression of surfactant protein mRNAs in the fibrotic lesions of Bleo-treated rats. KGF may play an important role in maintaining alveolar epithelium and repairing the damaged epithelium after lung injury. PMID- 9875146 TI - Localization of expression of human beta defensin-1 in the pancreas and kidney. AB - Defensins are antimicrobial peptides which play a key role in innate immunity. High levels of human beta defensin-1 (hBD-1) have previously been detected in the kidney and pancreas, but the cell-specific location of hBD-1 mRNA has not been determined. The expression of hBD-1 mRNA has been examined in fetal and adult pancreas and kidney by mRNA in situ hybridization. In fetal pancreas, hBD-1 expression was detected in the developing acini and in adult pancreas in the acini, but not in the pancreatic ducts. In both fetal and adult kidney, hBD-1 expression was detected in the collecting ducts and in the loops of Henle in adult kidney. Expression of hBD-1 mRNA in the pancreas and kidney from early development and in the acini of the adult pancreas, rather than in the pancreatic ducts, may indicate that in these tissues, hBD-1 fulfils physiological functions in addition to host defence. PMID- 9875147 TI - Proliferation of parenchymal epithelial cells enhanced in chronic pancreatitis. AB - This study was performed to determine whether pancreatic parenchymal epithelial cells in human chronic pancreatitis tissues retain a biologically significant capability to proliferate and, if so, within which epithelial compartment proliferation occurs. The techniques of immediate per-operative in vitro labelling with bromodeoxyuridine (BrdU) and conventional immunohistochemistry for Ki-67 antigen expression were used to identify proliferating cells. Concordance between the two techniques was confirmed in all tissues examined. In normal pancreas, proliferation was restricted to acinar epithelial cells, with no activity in the ductules. In chronic pancreatitis of both chronic obstructive and chronic calcifying types, the number of proliferating cells in the acini was significantly increased. A small population of proliferating cells was also apparent within ductules in chronic calcifying pancreatitis, but not in chronic obstructive pancreatitis. This investigation has shown that loss of parenchymal epithelium occurring in chronic pancreatitis is not caused by a primary failure of pancreatic 'stem-cell' proliferation, but is due to disproportionate attrition of differentiated parenchymal epithelial cells by a mechanism, possibly stromal in origin, which remains hitherto unidentified. The presence of proliferating ductular cells in chronic calcifying pancreatitis, but not chronic obstructive pancreatitis, suggests that distinct pathogenic processes may be operating in the former condition, which is classically regarded as secondary to ductal obstruction by stones, and in this single respect might be considered to be identical to chronic obstructive pancreatitis. Preservation of 'stem-cell' function supports the belief that regeneration of pancreatic parenchymal tissue could be a feasible proposition if biologically appropriate management strategies were developed to treat chronic calcifying pancreatitis. PMID- 9875148 TI - Studies on the innervation of human renal allografts. AB - Kidneys are innervated by a plexus of nerves around the renal artery, which is disrupted by transplantation. This is a report of a comparison of the nerves in human renal allografts and normal kidneys. There were many sympathetic ganglia around normal renal arteries but none around transplanted vessels, although equal numbers of ganglia were present in hilar tissues of normal and transplanted kidneys. An immunohistological study with an antibody to synaptophysin showed that the number of synapses in transplanted ganglia was severely reduced. Immunohistological studies on allograft kidneys using antibodies to various neurofilament proteins and the Schwann cell marker S100 showed a marked reduction in neurofilament proteins shortly after transplantation with subsequent partial recovery, and a lesser reduction in S100. Renal allografts have structurally abnormal innervation but are not completely denervated. PMID- 9875149 TI - Effect of prolonged exposure to oestradiol on subsequent LH secretion in ewes. AB - Abnormal follicles can produce oestradiol continuously for up to 20 days and this inhibits the hypothalamic-pituitary axis. The present experimental series was designed to determine the minimum exposure to high or low follicular phase concentrations of oestradiol that were required to exert inhibitory effects on LH surge secretion induced by additional oestradiol administered at the end of continuous exposure to oestradiol. Experiments were also included to establish whether the inhibitory effects of prolonged oestradiol were mediated at the pituitary, and whether the failure of response to the oestradiol challenge could be corrected by exposure to normal luteal phase patterns of progesterone. Treatment of ewes between 2 and 12 days with 1, 2 or 4 oestradiol implants (3 cm) totally blocked the subsequent normal LH surge in response to an oestradiol challenge in 45 of 52 ewes pretreated with oestradiol. In the seven ewes that did have an increase in LH, the response occurred at the expected time but was greatly reduced (14 versus 40 ng ml-1), and occurred only in ewes pretreated with oestradiol implants for 2 or 4 days. We were unable to establish a robust linear time-dose relationship, i.e., when ewes were treated with lower doses of oestradiol (one or two implants) for a reduced time (2, 4 or 8 days), there was random distribution of the 7 of 32 animals that had a reduced LH surge after oestradiol challenge (with four implants or 50 micrograms injection). The present study is the first to show that exposure for only 2-4 days to continuous oestradiol at late follicular phase concentrations can disrupt LH surge release. However, in oestradiol-treated ewes, LH secretion was provoked by high or low doses (0.5 mg or 0.5 microgram) of GnRH, although it was reduced by 50%, and a GnRH self-priming effect was still evident. All of these results suggest that inhibitory effects occur at the pituitary and hypothalamus. It remains to be confirmed whether the major effect is at the pituitary by reducing GnRH receptor or LH synthesis, or at the hypothalamus via inhibition of GnRH secretion. PMID- 9875150 TI - Effects of ageing and oestrogen on the localization of immunoglobulin-containing cells in the chicken ovary. AB - Immunoglobulins in the chicken ovary are important for transfer of immunity to chicks through the egg and for protection of the ovary from infection. The aim of this study was to examine the effects of ageing and oestrogen on the population of Ig-containing cells in the chicken ovary. The ovarian tissue of immature, young laying and old laying hens and that of immature birds treated with diethylstilboestrol (DES), progesterone or sesame oil (vehicle) was processed for paraffin wax sections. The sections were stained for IgG, IgM and IgA by an indirect immunostaining method and the population of cells positive for each Ig was analysed under a light microscope. The number of cells positive for IgG, IgM and IgA was significantly greater in the ovarian stromal tissue of young laying hens than in immature or old laying hens (P < 0.01). The number of IgG- and IgM positive cells in the thecal layer of primary follicles of young laying hens was significantly greater than that in immature and old laying hens (P < 0.01) and there were significantly more (P < 0.05) IgA-positive cells in young laying hens than in immature birds. The number of IgG-, IgM- and IgA-positive cells was significantly (P < 0.01) greater in both the stromal tissue and the thecal layer of DES-treated birds than in the vehicle-treated birds. Progesterone had no significant effect (P < 0.05) on the population of Ig-positive cells. These results indicate that the number of Ig-positive cells increases as chickens mature and decreases with ageing, and that oestrogen may be involved in this process. PMID- 9875151 TI - Regional influences of the fallopian tubes on the rate of boar sperm capacitation in surgically inseminated gilts. AB - Aliquots of ejaculated boar semen containing known numbers of spermatozoa were deposited into the caudal isthmus or rostral ampulla of the Fallopian tubes of gilts at, or immediately after, ovulation to assess regional influences on the rate of capacitation. Eggs were recovered during a second intervention 4, 5, 6 or 7 h after surgical insemination and were examined by phase-contrast microscopy. Results were obtained from ten animals in each of the 4-, 5- and 6-h groups and from eight animals in the 7-h group. With two exceptions, fertilized eggs were not recovered until 6 h after insemination into the isthmus, the proportion (45.6%) being significantly greater than the corresponding figure (1.4%) for ampullary insemination (P < 0.001). Similarly, the proportion of fertilized eggs recovered 7 h after insemination into the isthmus (58.7%) was significantly greater than after ampullary insemination (21.9%; P < 0.01). Numbers of spermatozoa associated with the zona pellucida remained low in all these instances, with mean figures per egg ranging from 0.3 to 3.8. Insemination into the isthmus gave a 1-2 h advantage in fertilization compared with insemination into the ampulla. Although relative rates of sperm cell progression to the site of fertilization may have contributed to this, there is strong evidence that rates of capacitation differ significantly in the respective portions of the Fallopian tube. Therefore, attention was focused on: (1) the viscous glycoprotein secretion in the caudal isthmus acting to remove seminal plasma from the sperm surface; and (2) the phase of sperm head binding to the isthmus epithelium. Gradients in local endocrine modulation by the adjacent ovary offer one explanation for the functional specialization of different regions of the Fallopian tubes. PMID- 9875152 TI - Spermadhesins of the AQN and AWN families, DQH sperm surface protein and HNK protein in the heparin-binding fraction of boar seminal plasma. AB - Heparin-binding proteins (designated BHB-2-BHB-9) were isolated from boar seminal plasma by affinity chromatography on heparin immobilized on polyacrylamide gel, followed by reverse phase HPLC. According to their N-terminal amino acid sequences, BHB-3-BHB-5 belong to the AQN family of spermadhesins and BHB-7-BHB-9 to the AWN family. BHB-6 is composed of two different proteins. The dominant protein (14 kDa) has the N-terminal amino acid sequence HNKQEGRDHD that is identical to the sequence of human semenogelin at positions 85-94. The minor proteins (16 and 17 kDa) belong to the AWN family of spermadhesins. The 14 kDa HNK protein does not crossreact with antibodies against AQN or AWN spermadhesins. BHB-2 also binds to the acrosome of boar epididymal spermatozoa but has the N terminal sequence DQH. Therefore, basic protein BHB-2 belongs to a new family of DQH sperm surface proteins that are homologous to the acidic proteins from bull and stallion seminal plasma, to the collagen binding domain II in fibronectin and to the leucocyte cell-cell adhesion regulator, but are not homologous to AQN or AWN spermadhesins. Nevertheless, anti-AQN-1 spermadhesin antibodies crossreact strongly with DQH protein. All boar heparin-binding proteins bind concanavalin A indicating their glycoprotein nature, which was proved by the detection of glucosamine and galactosamine residues in their molecules. Furthermore, spermadhesins interact with zona pellucida, protease inhibitors and a polyacrylamide derivative of heparin. Affinity chromatography experiments showed that the DQH protein bound to gelatin-agarose together with the AWN proteins and that the DQH protein and AQN-1 spermadhesin belong to the phosphoryl choline binding proteins. PMID- 9875153 TI - Expression of an inter-alpha-trypsin inhibitor heavy chain-like protein in the pig endometrium during the oestrous cycle and early pregnancy. AB - In pigs, changes in an unidentified endometrial glycoprotein, pGP30, are temporally associated with rapid trophoblast elongation and initial placental attachment on day 12 of gestation. Identification of endometrial pGP30 was undertaken through protein purification, NH2-terminal amino acid sequencing and cDNA sequencing of products generated through reverse transcription-polymerase chain reaction. Sequencing of 35 amino acids from the NH2-terminal end of pGP30 revealed that the 30 kDa glycoprotein is a cleavage product from the C-terminal region of inter-alpha-trypsin inhibitor heavy chain 4 (I alpha IH4), previously known as inter-alpha-trypsin inhibitor heavy chain-like protein. I alpha IH4 is unique compared with the three other inter-alpha-trypsin inhibitor heavy chains as it does not contain a binding site for bikunin that has serine protease inhibitory activity and is sensitive to cleavage by kallikrein. Endometrial gene expression of I alpha IH4 was detected during the oestrous cycle (days 0-18) and early pregnancy (days 10-18). Gene expression of I alpha IH4 appeared to be enhanced during the midluteal phase (days 12 and 15) of the oestrous cycle and the period of trophoblast attachment (days 12-18). Expression of I alpha IH4 was not detected in day 12 conceptus tissue mRNA. Endometrial expression of I alpha IH4 in pigs may function as an acute phase protein for protection of the uterus from the inflammatory response induced by conceptus attachment to the uterine epithelium. PMID- 9875155 TI - Sperm-oviduct epithelial cell monolayer co-culture: an in vitro model of sperm female tract interactions in a marsupial, the tammar wallaby (Macropus eugenii). AB - Oviduct epithelial cell (OEC) monolayers were prepared from the isthmic and ampullary parts of the oviducts of FSH-primed tammar wallabies. Co-culture experiments found that 50-60% of wallaby spermatozoa attached immediately to OEC monolayers, tracheal cell monolayer controls, and the surface of culture dishes with and without Matrigel coating. Spermatozoa were considered to be attached if they remained on the culture surface after rapidly pipetting the co-culture medium five times. The percentages of attached and unattached spermatozoa were calculated from the number of spermatozoa recovered in the agitated supernatant. After 2 h co-culture the percentage of attached spermatozoa rose to 60-80%. After 6 h co-culture the number of spermatozoa attached to OEC monolayers derived from the oviductal isthmus remained high and only a small percentage were recovered in the agitated supernatant (unattached spermatozoa 3.85 +/- 0.76%, P = 0.67). However, after 6 h co-culture of spermatozoa with OEC monolayers derived from the ampulla and with the controls the percentage of attached spermatozoa declined significantly (unattached spermatozoa: ampullary monolayer 23.08 +/- 4.80%, P < 0.01; tracheal monolayer 23.23 +/- 5.18%, P < 0.01; Matrigel 27.23 +/- 7.76%, P < 0.01; plastic surface 28.19 +/- 5.30%, P < 0.01). After 6 h co-culture with ampullary and isthmic OEC monolayers, the percentage motility of both attached and unattached spermatozoa was maintained at 64.00 +/- 1.90% and 56.66 +/- 3.18% and 62.00 +/- 3.11% and 52.00 +/- 2.43%, respectively, and was then maintained at > or = 35% after 24 h incubation. In the controls, that is, tracheal monolayer and Matrigel, the motility of attached spermatozoa declined rapidly to 48.66 +/- 2.15% and 33.63 +/- 8.66%, respectively, at 6 h, and all spermatozoa were immotile after 24 h incubation. However, the motility of unattached spermatozoa in the controls (tracheal monolayer and Matrigel) was maintained at 57.33 +/- 3.00% and 34.54 +/- 9.27%, respectively, until 6 h and then declined rapidly, and all spermatozoa were immotile after 24 h incubation. Co-culture of wallaby spermatozoa with OEC monolayers also induced acrosomal modifications that were followed by acrosomal loss. At 6 h incubation 38.92 +/- 3.98% of spermatozoa on ampullary OEC monolayers and 36.50 +/- 3.81% spermatozoa on isthmic OEC monolayers had shed their acrosome. Acrosomal loss during co-culture with both isthmic and ampullary OEC monolayers was significantly (P < 0.01) greater than that observed on tracheal epithelial monolayer (24.42 +/- 1.90%, P < 0.01), Matrigel (20.70 +/- 2.71%, P < 0.01) and plastic (15.54 +/- 2.49%, P < 0.01). Co culturing spermatozoa with OEC monolayers also induced a transformation from streamlined orientation of sperm head and tail to T-shaped (thumbtack) orientation in a small number (10-15%) of motile spermatozoa after 6 h incubation (data not shown). The significance of these results in relation to the role of the oviduct in sperm capacitation is discussed. PMID- 9875154 TI - Effects of age and gonadal steroids on the localization of antigen-presenting cells, and T and B cells in the chicken oviduct. AB - The effects of age and gonadal steroids on the localization of immunocompetent cells, including antigen-presenting cells that contain the major histocompatibility complex (MHC) class II antigen, and T and B cells in the chicken oviduct were studied. Oviductal tissues were collected from laying and immature hens treated with diethylstilboestrol (an analogue of oestrogen) or progesterone. Cryostat sections of the tissues were immunostained for MHC class II, CD3 (T-cell antigen) and Bu-1 (immature B-cell antigen), and examined under a light microscope and an image analysis system. MHC class II+, CD3+ and Bu-1+ cells were observed in the mucosal epithelium and stromal connective tissue of both the laying and immature hens. MHC class II+ cells in the oviductal stroma appeared in association with oviductal development during sexual maturation and increased with ageing thereafter. The infiltration of CD3+ and Bu-1+ cells into the oviductal tissues increased in young laying hens compared with immature hens and decreased in old laying hens compared with young laying hens. Diethylstilboestrol increased the population of MHC class II+ and CD3+ cells in the stroma of the infundibulum and vagina, but had no significant effect on the population of Bu-1+ cells in the oviduct of immature hens. Progesterone increased the population of CD3+ cells in the stromal tissue of oviductal segments from all hens, and of Bu-1+ cells in the mucosal epithelium of the infundibulum and magnum, but had little effect on the frequency of MHC class II+ cells in the oviduct of immature hens. There were typically more immunocompetent cells in the infundibulum and vagina than in the other oviductal segments in laying hens and immature hens treated with sex steroids. These results suggest that local immunity in the chicken oviduct is enhanced during sexual maturation and possibly decreases during ageing. Gonadal steroids may play a significant role in the regulation of local immunity in the oviduct. The effects of oestrogen and progesterone on the influx of these immunocompetent cells into the oviduct differs among cell types and oviductal segments. PMID- 9875156 TI - Ovarian androstenedione production is enhanced by insulin during the period of delayed ovulation in a vespertilionid bat, Scotophilus heathi. AB - The aim of the present study was to evaluate the role of insulin in the production of high concentrations of androstenedione in the ovary of Scotophilus heathi during the period of delayed ovulation. The concentrations of serum insulin were found to be high during November and December and declined significantly before ovulation. A significant correlation was observed between circulating insulin concentration and body weight as well as with serum androstenedione concentrations. Autoradiographic study demonstrated specific accumulation of 125I-labelled insulin in the ovarian thecal and interstitial cells. Insulin increased both basal and hCG-stimulated androstenedione production by the ovary in vitro during November. These results indicate that insulin may be involved in inducing high androstenedione synthesis by the ovary and thus may be indirectly responsible for inducing delayed ovulation in S. heathi. PMID- 9875157 TI - Regulation of prostaglandin F2 alpha and E receptor mRNA by prostaglandin F 2 alpha in ovine corpora lutea. AB - Prostaglandins regulate many physiological functions, including reproduction, by binding to specific plasma membrane receptors. In this study we evaluated the regulation of PGF2 alpha (FP) and PGE (EP3 subtype) receptors in ovine corpora lutea. In the first study, tissue distribution of FP and EP3 receptors was evaluated in 13 ovine tissues. FP receptor mRNA was present in 100-fold higher concentration in corpora lutea than in other tissues. Similarly, [3H]PGF2 alpha binding was much greater in luteal plasma membranes than in membranes from other tissues. In contrast, EP3 receptor mRNA was more uniformly distributed, with high concentrations in adrenal medulla, inner myometrium, kidney medulla and heart. The distribution of [3H]PGE1 binding was generally similar to EP3 mRNA, with the exception that ovarian stroma, endometrium and outer myometrium had high [3H]PGE1 binding but low concentrations of EP3 receptor mRNA. The second study evaluated the action of PGF2 alpha on luteal mRNA encoding FP and EP3 receptors. Ewes had PGF2 alpha or saline infused into the ovarian artery and corpora lutea were removed at 0, 1, 4, 12 and 24 h. FP receptor mRNA decreased by 50% at 12 and 24 h after infusion with PGF2 alpha, whereas EP3 mRNA was unchanged. Treatment of large luteal cells with PGF2 alpha, phorbol didecanoate (protein kinase C activator), or ionomycin (calcium ionophore) decreased FP receptor mRNA after 24 h (P < 0.05). Glyceraldehyde 3-phosphate dehydrogenase mRNA was not changed by any treatment. These results show that EP3 receptors are expressed in many tissues and expression is not regulated by PGF2 alpha. In contrast, FP receptors are primarily expressed in corpora lutea and expression is inhibited by PGF2 alpha. PMID- 9875158 TI - Immunostimulatory effects of pig seminal proteins on pig lymphocytes. AB - Pig seminal proteins PSP-I and PSP-II are major protein components of boars' ejaculate and are present as heterodimers (PSP-dimer) in seminal plasma. These proteins were examined for their ability to modulate pig lymphocyte activity in vitro in mitogen-induced lymphocyte proliferation assays and in one-way mixed lymphocyte reactions. Pig lymphocytes were cultured with phytohaemagglutinin, concanavalin A, or pokeweed mitogen (PWM) in the presence or absence of pig seminal proteins and the amount of cellular [3H]thymidine was used as an indication of proliferation. In the absence of mitogens, none of the three pig seminal proteins affected lymphocyte proliferation suggesting that these proteins are not antigenic or mitogenic. PSP-dimer enhanced lymphocyte proliferation induced by PWM (156-227%, P < 0.05) in a concentration-dependent manner, but had no effect on phytohaemagglutinin- or concanavalin A-induced proliferation. PSP-I enhanced (127-185%, P < 0.05) phytohaemagglutinin-induced proliferation. PSP-II augmented (130-240%, P < 0.05) lymphocyte proliferation induced by concanavalin A and PWM. Lymphocytes from gilts were significantly more responsive to concanavalin A- and PWM-induced lymphocyte proliferation in the presence of PSP-I compared with boars (concanavalin A: gilts 131%, boars 91%; PWM: gilts 188%, boars 134%; P < 0.05). In the mixed lymphocyte reaction, pretreating stimulating cells with increasing concentrations of PSP-I or PSP-II elicited a 400% concentration-dependent increase (P < 0.01) in lymphocyte proliferation. The abundance of pig seminal proteins in boar seminal plasma, their ability to enhance lymphocyte proliferation, and their previously reported ability to bind to lymphocytes suggest that these proteins are immunostimulatory and supports the hypothesis that they modulate uterine immune activity to ensure reproductive success. PMID- 9875159 TI - Testosterone, LH and FSH episodic secretory patterns in GnRH-immunized bulls. AB - The objective was to determine: (i) the secretory patterns of LH, FSH and testosterone in bulls, and the temporal relationships between the pulses of these hormones; and (ii) the effect of GnRH immunization on these parameters. Friesian bulls (n = 72) were given a primary immunization on day 0 (10-week-old) and a booster immunization on either day 28 (n = 36) or day 56 (n = 36) against either a GnRH-human serum albumin (HSA) conjugate (n = 48) or HSA (n = 24; control). On the basis of GnRH antibody titres at a plasma dilution of 1:160, 1 week after booster immunization, 12 GnRH-immunized and six control bulls from each booster immunized group were selected and allocated to one of three groups: control bulls and bulls with medium and high antibody titres (0.3, 32 and 51% binding, respectively; pooled SED 4.3%). Blood samples were taken from these animals (n = 36) every 15 min for 8 h on three occasions: (i) 2 weeks after booster immunization when bulls were 4-5 months of age (prepubertal); (ii) at 7 months of age (peripubertal); and (iii) at 11 months of age (post-pubertal). Data were analysed by PULSAR, ANOVA and chi-squared test. The mean antibody titre of the high antibody titre group was greater (P < 0.05) than that of the medium antibody titre group in prepubertal bulls only, but the mean antibody titres of both antibody titre groups were greater (P < 0.05) than that of the control bulls at all times. The frequency and amplitude of LH and FSH pulses in the control bulls were greater (P < 0.05) during prepuberty than after puberty. The frequency, amplitude and duration of LH pulses were greater (P < 0.05) in control bulls than those in medium and high antibody titre bulls at prepuberty. The mean and basal concentrations of FSH and the amplitude and duration of FSH pulses were greater (P < 0.05) in the control bulls than in the high antibody titre bulls at prepuberty. The frequency of testosterone pulses was greater (P < 0.05) in the control bulls than in the medium and high antibody titre bulls at peripuberty. The mean and basal concentrations and pulse amplitude of testosterone were greater (P < 0.05) in high antibody titre bulls than in control bulls after puberty. There was a close temporal relationship between LH and FSH (62.5% of LH pulses were followed by FSH pulses within 75 min) at prepuberty in the control bulls but there was no relationship after puberty. The opposite trend occurred in the high antibody titre bulls, that is, there was no relationship between LH and FSH at prepuberty but there was a close temporal relationship after puberty. The temporal relationship between LH and testosterone was closest at peripuberty (86.7%) in the control bulls, but increased in the high antibody titre bulls from 0% at prepuberty to 57.1% after puberty. In summary, there was an age-related decrease in LH and FSH pulse frequency and amplitude and also in the temporal relationships between these hormones in control bulls. Prepubertal GnRH immunization had a suppressing effect on the pulsatile release of LH, FSH and testosterone before but not after puberty. The presence of high amplitude testosterone pulses in the GnRH-immunized bulls after puberty indicates that the long-term tonic release of LH may be sufficient to initiate a late pubertal-type increase in testosterone concentrations. PMID- 9875160 TI - Semen parameters and electron microscope observations of spermatozoa of the red wolf, Canis rufus. AB - Semen parameters were evaluated on ejaculates of a captive population of red wolves (Canis rufus) sampled over two consecutive mating seasons. A total of 31 samples from 15 animals yielded mean sperm motility of 69.6 +/- 19.4%, mean sperm density of 131 +/- 124 x 10(6) ml-1, mean total number of spermatozoa of 470 +/- 465 x 10(6) and mean percentage morphologically abnormal spermatozoa of 35 +/- 11.8%. Restricting the data to animals sampled three times or more or limiting the samples to proven breeders resulted in statistically non-significant differences in these numbers (P < 0.05). When compared with data from other canines the seminal parameters of red wolves are at the lower extremes of the range. In particular the proportion of morphologically abnormal spermatozoa (35%) is approximately twice that seen in other canine species. Light microscopic analysis of abnormal forms revealed that almost half (45%) were bent defects, another 40% were secondary defects (coiled, detached and immature) and 15% were primary defects. Electron microscopy confirmed the presence of substantial numbers of morphologically abnormal forms including double-headed and double flagellar cells, bent or kinked forms especially in the neck region, acrosomal abnormalities and bizarre spermatids. Approximately one-third of the samples also showed the presence of white blood cells, in some cases demonstrating sperm phagocytosis (spermophagy). These results are consistent with the concept of declining sperm parameters associated with restricted gene pools in numerically limited populations. However, alternative explanations are also explored. PMID- 9875161 TI - Localization of glucose-6-phosphatase activity and carbohydrates in boar caput epididymal principal cells. AB - Unidentified tubulovesicular profiles have been reported in the apical cytoplasm of boar caput epididymal principal cells in addition to vesicles considered to be involved in endocytosis and secretion. The main aim of the present study was to clarify the character of these organelles and to differentiate them from the endocytic apparatus. Glucose-6-phosphatase (G6Pase) activity was determined as the reporter enzyme of the endoplasmic reticulum (ER) and phosphotungstic acid was used to visualize carbohydrate moieties in both the proximal and distal caput. Phosphotungstic acid revealed the glycocalyx of the endocytic apparatus, which was similar in both regions studied, and also stained specific granules of the proximal caput. Glucose-6-phosphatase showed the tubulovesicular profiles to be sparsely granulated ER that was poorly developed in the proximal caput and very abundant in the apical cytoplasm of the distal caput principal cells. The function of such large amounts of sparsely granulated ER with corresponding G6Pase activity in caput epididymal principal cells is unknown. PMID- 9875162 TI - Physiological responses of hens divergently selected on the number of chicks obtained from a single insemination. AB - A series of experiments was conducted in domestic fowl to investigate the consequences of five generations of divergent selection for increased (L+) or decreased (L-) numbers of hatched chicks. After artificial insemination with pooled ejaculates within the same line (L+ males x L+ hens or L- males x L- hens), significant differences were observed between L+ and L- hens for mean fertility rates (L+ 94.8%, L- 70.2%, P < 0.0001) and for effective and maximum duration of fertility (P < 0.00001). A comparison of the overall laying performance and shell quality between the two selected lines showed that L- hens laid fewer eggs than L+ hens (P < 0.00001) and L- eggs had poorer shell quality (shell breaking strength) than L+ eggs (P < 0.00001). These observations were associated with significantly higher percentages of early embryo death in eggs from L- hens compared with L+ hens. Another series of experiments revealed the presence of larger initial populations of spermatozoa in the sperm storage tubules as well as in the perivitelline layer of eggs from L+ hens. The populations of spermatozoa in the sperm storage tubules of commercial laying hens inseminated with pooled semen samples from L+ males was compared with those in hens inseminated with samples from L- males to determine whether the variations in oviductal sperm storage between the two lines were male dependent. No significant differences between the populations of spermatozoa present in the sperm storage tubules of either group of hens could be detected at any of the intervals examined after insemination (days 1, 3 and 10). Finally, an experiment conducted on hens originating from the two selected lines indicated that the utero-vaginal junction of L+ hens contained significantly more sperm storage tubules compared with L- hens (P < 0.01). It is concluded that selection based on overall reproductive performance modifies the number of eggs capable of developing viable embryos and also influences the efficacy of initial sperm storage by increasing or altering the population of sperm storage tubules located in the utero-vaginal junction. Such changes have major consequences on the duration of the fertile period, which in avian species is directly dependent on both the actual population of spermatozoa stored in the oviduct and on their rate of release from the storage sites. PMID- 9875163 TI - Regulation of urokinase- and tissue-type plasminogen activator by relaxin in the uterus and cervix of the prepubertal gilt. AB - Changes in plasminogen activator are associated with the reproductive tissue remodelling that occurs during growth. Given the trophic effects of relaxin on the pig uterus and cervix, the present study was designed to examine the impact of relaxin on urokinase and tissue-type plasminogen activator (uPA and tPA) protein and activity in the uterus and cervix of prepubertal pigs. After relaxin administration in vivo to induce growth of the immature uterus and cervix, plasminogen activator activity was measured in uterine flushes and uterine and cervical tissue using a chromogenic substrate assay. Immunoreactive uPA and tPA protein in uterine flushes and uterine and cervical tissue was detected by western blotting. Urokinase plasminogen activator activity was significantly higher (P < 0.05) in uterine flushes from relaxin-treated animals than in controls. However, there was no change in uterine flush tPA activity or protein in response to in vivo relaxin treatment. There was no evidence for acid-labile inhibitors of plasminogen activator in uterine flushes of any of the animals. Cell-associated uterine tissue uPA and tPA activity, as well as protein, were similar in relaxin-treated and control prepubertal pigs. In the cervix, cell associated tPA activity decreased significantly (P < 0.05) in relaxin-treated animals, while cervical uPA activity was unchanged. These results support the view that at least one means by which relaxin promotes pig uterine growth is by increasing uterine secretion of uPA. In addition, these studies suggest that relaxin administration in vivo to prepubertal gilts has tissue-specific effects with respect to plasminogen activator. PMID- 9875164 TI - Inhibition by oestradiol of oxidative stress-induced apoptosis in pig ovarian tissues. AB - Oxidative stress-induced apoptotic cell death has been implicated in the mechanisms of corpus luteum regression and follicular atresia. The objective of this investigation was to determine whether the antioxidant, oestradiol influenced apoptosis in pig luteal and follicular tissues exposed in vitro to hydrogen peroxide. Immunolocalization of fragmented DNA (a biochemical marker of apoptosis) in large steroidogenic luteal and granulosa cells was inhibited by oestradiol. Protection against apoptosis required relatively high concentrations of oestradiol (> or = 40 pg ml-1 incubation media) and was largely unmitigated by actinomycin D (suggesting a predominantly receptor-independent nongenomic mode of action). Ovarian cells were not protected by other (nonaromatizable) steroid hormones. It is suggested that oestradiol functions at the ovary as a reactive oxygen scavenger during pregnancy-mediated luteal rescue and folliculogenesis. PMID- 9875165 TI - Steroid contents of and steroidogenesis in vitro by the developing gonad and mesonephros around sexual differentiation in fetal sheep. AB - The aim of the present study was to establish whether the steroids, progesterone, androstenedione, testosterone and oestradiol, were present in the mesonephric gonadal complex of female and male sheep fetuses around sexual differentiation (that is, from day 28 to day 45 of gestation, with sexual differentiation occurring at approximately day 32). A second aim was to test whether the mesonephric-gonadal complex, mesonephros (days 35-45 only) and gonad (days 35-45 only) were capable of steroid synthesis in vitro. The steroid contents in the mesonephric-gonadal complex were not detectable before sexual differentiation. However, from day 35 of gestation onwards, the mesonephric-ovarian complex contained mainly oestradiol and the mesonephric-testicular complex contained mainly testosterone: from day 35 until day 45 the increase in content of these two steroids exceeded the increase in the mass of tissue by more than fivefold. From day 40 to day 45 of gestation, the contents of the other steroids in the pathways to oestradiol increased progressively in both sexes but more in parallel with the increase in tissue mass. In contrast to the steroid contents in the tissue at recovery, the mesonephric-gonadal tissue from both sexes in tissue culture was able to synthesize most steroids before and after sexual differentiation and also to metabolise supplementary androstenedione to oestradiol. These findings suggest that many, if not all, of the steroidogenic enzymes in the pathway from cholesterol to oestradiol are present before sexual differentiation. Most of the aforementioned steroids were present in detectable amounts in isolated mesonephros and gonad of both sexes after sexual differentiation. Moreover, for both the isolated mesonephros and gonad, there were increases in the mean contents of most steroids after culture relative to the contents in the tissues at recovery. These data suggest that the mesonephros, as well as the gonad, in both sexes is capable of synthesizing steroid. It is concluded that, in the sheep fetus, the female and male gonads are steroidogenically active after sexual differentiation, that the steroidogenic enzymes develop before sexual differentiation, and that the mesonephros is a site of steroid synthesis. PMID- 9875166 TI - Sperm transport in the reproductive tract of female zebra finches (Taeniopygia guttata). AB - Aspects of sperm transport in the oviducts of female zebra finches were examined by recording the decline in the number of spermatozoa on the outer perivitelline layer of successively laid eggs. Data from a single clutch of eggs from 32 females indicates that the mean per capita rate of loss of spermatozoa was estimated to be 0.0170 +/- 0.002 SEM spermatozoa h-1. However, individual females showed no consistency in the rate at which spermatozoa were lost from their oviduct over five successive clutches. Models of the mechanism of sperm competition in birds assume that the rate of loss of spermatozoa does not differ between inseminations made before or after the start of egg laying. This assumption was found to be valid: the instantaneous per capita rate of loss of spermatozoa did not differ significantly between females inseminated either before (0.01445 +/- 0.0028 SEM spermatozoa h-1) or after (0.01674 +/- 0.0023 SEM spermatozoa h-1) the onset of oviposition. The rate of sperm transport through the infundibulum was determined to be slower than that between the utero-vaginal sperm storage tubules and the infundibulum by comparing the number of spermatozoa associated with the perivitelline layers of eggs laid after a day on which no eggs were laid. Eggs with < 20 spermatozoa on the outer perivitelline layer were found to have a 50% probability of being infertile. The results are compared with data for domestic fowl and turkeys. PMID- 9875167 TI - Effect of immunization against the amino-terminal peptide (alpha N) of the alpha 43-subunit of inhibin on follicular atresia and expression of tissue inhibitor of matrix metalloproteinase (TIMP-1) in ovarian follicles of sheep. AB - Ewes actively immunized against alpha N, the N-terminal peptide of inhibin alpha 43 precursor, have lowered fertility associated with ovulation failure, restricted tissue remodelling and reduced matrix metalloproteinase-2 activity in the follicular fluid at the time of expected ovulation. This could be due to altered ratios of matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase (TIMP-1), or to the onset of atresia in antral follicles destined to ovulate. The objectives of the present study were to investigate the effects of immunization against alpha N on the localization of TIMP-1 in ovine follicles, and on follicular growth and atresia in the follicular phase. Ewes were either immunized against alpha N or remained as controls and the ovaries were removed before (0, n = 4) and at 12 h (n = 4) and 24 h (n = 4) after hCG administration in a synchronized follicular phase, 48 h after removal of intravaginal pessaries. Observations were made on a single section taken through the largest follicle present in the ovaries of each ewe. There were no healthy antral follicles > 1 mm in immunized ovaries (0/29) compared with controls (16/31) (P < 0.001), whereas the proportion of healthy antral follicles < 1 mm was the same in each group (9/19 versus 5/12). TIMP-1 immunoactivity was localized in large luteal cells, smooth muscle and endothelial cells, and in all antral follicles, including oocytes. At the time of hCG administration, no TIMP-1 immunoreactivity was detected in the apical region of the follicular wall of large follicles (> 6 mm) compared with the rest of the follicle wall, but staining appeared in the apical granulosa layer 24 h later. In newly formed corpora lutea, TIMP-1 expression was found along the invaginating vascular layer. There was no effect of immunization on the patterns of TIMP-1 immunoreactivity, suggesting that changes in TIMP-1 are not involved in the effects of alpha N. These data are consistent with a paracrine role for alpha N in the selection and atresia of antral follicles, and for TIMP-1 in tissue reorganization and steroidogenesis at the time of ovulation. PMID- 9875169 TI - Ubiquitin and apoptosis in the corpus luteum of the marmoset monkey (Callithrix jacchus). AB - The polypeptide ubiquitin covalently binds to cytoplasmic proteins and marks them for proteolytic degradation. Ubiquitin is upregulated during apoptosis in some systems. Apoptosis increases during luteolysis but it is not known whether ubiquitin is expressed in regressing corpora lutea. Marmoset ovaries were removed on day 10 of the luteal phase from animals that had received either no treatment, treatment with the PGF2 alpha analogue cloprostenol 24 h earlier, or treatment with the GnRH antagonist antarelix for either 24 or 48 h before ovary collection. Ubiquitin was localized on ovarian sections by immunocytochemistry, and oligonucleosome formation characteristic of apoptosis was examined in isolated corpora lutea by electrophoresis of extracted [32P]DNA. Oligonucleosome formation was low in midluteal corpora lutea on day 10 but increased after induced luteal regression with PGF2 alpha and GnRH antagonist. Nuclear ubiquitin immunoreactivity was found in 1.66 +/- 0.66 steroidogenic cells and cytoplasmic staining was found in 0.4 +/- 0.3 steroidogenic cells (per x 40 field of view) in midluteal phase corpora lutea on day 10. Luteolytic induction with PGF2 alpha significantly increased the number of cells exhibiting cytoplasmic immunoreactivity to 12.24 +/- 1.6 (P < 0.05). Ubiquitin immunoreactivity was not observed after GnRH-induced luteal regression. Apoptotic oligonucleosome formation was found after induced luteal regression with both PGF2 alpha and GnRH antagonist, but ubiquitin upregulation only occurred after PGF2 alpha-induced regression. These results indicate that ubiquitin expression is not specific for luteolysis and is not an indicator of luteal apoptosis, but that the polypeptide does play a role in luteal cellular responses to PGF2 alpha. PMID- 9875168 TI - The role of nitric oxide in the process of implantation in rats. AB - Nitric oxide (NO) plays an important role in cell signalling in many physiological systems, including reproduction. During pregnancy, oestrogen modulates uterine NO generation, and NO may play an intermediary role in the oestrogen-mediated effects on the uterus. Since oestrogen is actively involved in inducing endometrial receptivity to support the process of implantation, the role of NO in the process of implantation in rats was investigated. N omega-nitro-L arginine methyl ester (L-NAME), an inhibitor of NO synthase (NOS), was administered in utero with or without sodium nitroprusside (SNP), a generator of NO, on different days during the preimplantation phase of gestation. The status of gestation in respect of implantation failure, endometrial receptivity and embryo development were assessed. L-NAME was administered at various doses (2-5 mg per uterine horn) and on different days of pregnancy (days 2-6 of pregnancy) to optimize the pregnancy terminating dose (absence of implantation site on day 8 of pregnancy) and the effective day of treatment. L-NAME led to failure of implantation when administered at 2.5 mg per uterine horn on day 3 of pregnancy. The characteristic preimplantation permeability changes in the uterus were significantly attenuated and embryo growth was retarded. The L-NAME-mediated effects were significantly reversed when SNP (100 micrograms) was co-administered with L-NAME. These findings suggest a role for NO in the process of implantation. The possible mechanism by which inhibition of the NO-NOS system may interfere with implantation is discussed. PMID- 9875170 TI - Protein synthesis and secretion by the epididymis of the brushtail possum, Trichosurus vulpecula. AB - In this study of brushtail possums, proteins present in epididymal fluid and not present in blood plasma and those that become associated with spermatozoa as they pass along the tract were investigated. At least 19 proteins were present in epididymal fluid in the various regions of the tract that were not detected in serum. Some of these may be present on the sperm plasmalemma. Six proteins were extracted from caput spermatozoa (M(r) 117,000, 103,000, 87,500, 85,000, 62,000 and 33,000) that did not appear in the caudal sperm extracts. Eight proteins (M(r) 50,000, 49,000, 32,000, 27,000, 26,500, 25,000, 24,500 and 18,000) were localized to caudal sperm extracts. These findings suggest that some sperm proteins are lost or modified, whereas others are added to the sperm plasma membrane during epididymal transit. In vitro incorporation of [35S]methionine by the epididymal tissue showed that the distal caput and corpus are the most active regions in the synthesis and secretion of proteins. Four caudal epididymal proteins (M(r) 72,000, 31,000, 26,500 and 21,000) were partially sequenced. Those of M(r) 31,000 and 26,500 had the same N-terminal amino acid sequence suggesting post-translational modification of the same protein; they showed homology to a retinoic acid-binding protein. The protein of M(r) 72,000 was found to be homologous to alpha-fetoprotein, whereas the protein of M(r) 21,000 showed no significant homology to any protein in the database. These results show that the lumen of the epididymis has many proteins that are not present in the blood, some of which appear to become associated with spermatozoa during epididymal transit. PMID- 9875171 TI - The upside and downside of cultural diversity. PMID- 9875172 TI - Grapefruit and medication. PMID- 9875173 TI - Is laughter the best medicine? PMID- 9875174 TI - Technology scorecard. Wound care science at the crossroads. PMID- 9875175 TI - Spinal injuries. PMID- 9875177 TI - Hospice care. PMID- 9875178 TI - Rebels with a cause. PMID- 9875179 TI - The therapeutic use of technology. PMID- 9875180 TI - Emergency! Hyponatremia. PMID- 9875181 TI - Helping our hidden youth. PMID- 9875182 TI - Rheumatoid arthritis. PMID- 9875183 TI - The benefits of visualization. PMID- 9875184 TI - Nursing care and patient satisfaction. PMID- 9875185 TI - Magnetic nursing. PMID- 9875186 TI - The effectiveness of mycophenolate mofetil in refractory pyoderma gangrenosum. PMID- 9875187 TI - Snake oil for the 21st century. AB - Dermatology has been associated with quackery for at least a century. The dictionary defines a quack as "a pretender to medical knowledge or skill; ignorantly or falsely pretending to cure." The term quack is derived from quacksalver, or one who quacks like a duck in promoting his salves. Quacksalvers hacked many potions, including snake oil, with claims that it cured everything from dermatitis to rheumatism. With the current promulgation of skin "products" and their promotion and even sale by dermatologists, and the use of treatments of no proven efficacy, this association between dermatology and quackery is set to continue well into the 21st century. The list of offending treatments includes silicone gel sheets and onion extract cream (Mederma) for keloids, alpha-hydroxy acid creams and peels, topical ascorbic acid and phytonadione, "laser resurfacing," and cimetidine for warts, to name only a few. PMID- 9875188 TI - Bringing evidence to the clinic. PMID- 9875189 TI - Treatment of lichen planus. An evidence-based medicine analysis of efficacy. AB - OBJECTIVE: To critically appraise the body of literature concerning treatment of lichen planus (LP). DESIGN: Review of MEDLINE and BIOSIS databases to identify articles published with at least an English abstract before March 1998 that examined treatment of LP. MAIN OUTCOME MEASURES: Forming a primary database on which most recommendations are based. We thus selected 83 clinical trials or small series of patients in the medical literature that referenced clinical data on patients treated for LP. RESULTS: There are no large randomized trials with definitive results in the medical literature examining the efficacy of the various drugs or physical treatments of LP. There are only 3 level B trials (small randomized trials with uncertain results because of moderate to high alpha or beta error) that address efficacy of treatment in LP, i.e., 1 with acitretin in cutaneous LP and 2 with topical corticosteroids in mucosal LP. The remainder of the published trials are observational and are not always prospective. Many of the recommendations of the experts are based on their personal experience. CONCLUSIONS: Although LP may be associated with substantial morbidity and altered quality of life, especially the erosive mucosal LP, definitive clinical trials have not been performed. Acitretin is the first-line therapy in cutaneous LP. The efficacy of systemic corticosteroids and psoralen plus UV-A therapy has not been established with a high level of proof. Topical corticosteroids are the first line therapy in mucosal erosive LP. Other treatments, such as topical cyclosporine or extracorporeal photochemotherapy, remain to be evaluated. European-US cooperation is warranted to perform large randomized controlled trials in cutaneous and mucosal LP. PMID- 9875190 TI - Nonsurgical repigmentation therapies in vitiligo. Meta-analysis of the literature. AB - OBJECTIVE: To assess the effectiveness and safety of nonsurgical repigmentation therapies in localized and generalized vitiligo by means of a meta-analysis. DATA SOURCES: Computerized searches of bibliographic databases, a complementary manual literature search, and contacts with researchers and pharmaceutical firms. STUDY SELECTION: Predefined selection criteria were applied to both randomized and nonrandomized controlled trials. DATA EXTRACTION: Two investigators independently assessed the articles for inclusion. When there was a disagreement, a third investigator was consulted. DATA SYNTHESIS: Sixty-three studies were found on therapies for localized vitiligo. Of these, 10 of 11 randomized controlled trials and 29 of 110 patient series were included. One hundred seventeen studies on therapies for generalized vitiligo were found. Of these, 10 of 22 randomized controlled trials and 46 of 231 patient series were included. Among randomized controlled trials on localized vitiligo, the pooled odds ratio vs placebo was significant for topical class 3 corticosteroids (14.32; 95% confidence interval [CI], 2.45-83.72). In the patient series, topical class 3 and class 4 corticosteroids carried the highest mean success rates (56% [95% CI, 50%-62%] and 55% [95% CI, 49%-61%], respectively). Side effects were reported mostly with topical psoralen and intralesional and class 4 corticosteroids. In the randomized controlled trials on generalized vitiligo, the odds ratio vs placebo was significant for oral methoxsalen plus sunlight (23.37; 95% CI, 1.33-409.93), oral psoralen plus sunlight (19.87; 95% CI, 2.37-166.32), and oral trioxsalen plus sunlight (3.75; 95% CI, 1.24-11.29). In the series, the highest mean success rates were achieved with narrowband UV-B (63%; 95% CI, 50%-76%), broadband UV-B (57%; 95% CI, 29%-82%), and oral methoxsalen plus UV-A therapy (51%; 95% CI, 46% 56%). Oral methoxsalen plus UV-A was associated with the highest rates of side effects. No side effects were reported with UV-B therapy. CONCLUSIONS: Class 3 corticosteroids and UV-B therapy are the most effective and safest therapies for localized and for generalized vitiligo, respectively. PMID- 9875191 TI - A systematic review of autologous transplantation methods in vitiligo. AB - OBJECTIVE: A systematic review of the effectiveness, safety, and applicability of autologous transplantation methods in vitiligo. DATA SOURCES: Computerized searches of bibliographical databases, a complementary manual literature search, and contacts with researchers and pharmaceutical firms. STUDY SELECTION: Predefined selection criteria were applied to all studies found. DATA EXTRACTION: Two investigators independently assessed the articles for inclusion. When there was a disagreement, a third investigator was consulted. DATA SYNTHESIS: Sixty three studies were found, of which 16 reported on minigrafting, 13 on split thickness grafting, 15 on grafting of epidermal blisters, 17 on grafting of cultured melanocytes, and 2 on grafting of noncultured epidermal suspension. Of these, 39 patient series were included. The highest mean success rates (87%) were achieved with split-skin grafting (95% confidence interval, 82%-91%), and epidermal blister grafting (87%) (95% confidence interval, 83%-90%). The mean success rate of 5 culturing techniques varied from 13% to 53%. However, in 4 of the 5 culturing methods, fewer than 20 patients were studied. Minigrafting had the highest rates of adverse effects but was the easiest, fastest, and least expensive method. CONCLUSIONS: Because no controlled trials were included, treatment recommendations should be formulated with caution. Split-thickness and epidermal blister grafting can be recommended as the most effective and safest techniques. No definite conclusions can be drawn about the effectiveness of culturing techniques because only a small number of patients have been studied. The choice of method also depends on certain disease characteristics and the availability of specialized personnel and equipment. PMID- 9875192 TI - How often does oral treatment of toenail onychomycosis produce a disease-free nail? An analysis of published data. AB - OBJECTIVE: To analyze studies on oral treatment of toenail onychomycosis so as to aid clinicians and patients in making informed decisions. DATA SOURCES: Studies dealing with treatment of toenail onychomycosis were identified by means of 2 MEDLINE search strategies. One was a title search using the word "toenail"; the other search used the combined Medical Subject Headings "onychomycosis" and "therapy". STUDY SELECTION: Articles were read to ascertain that they (1) described results in toenails, (2) used both culture and microscopy, and (3) included a clinical evaluation. Not included were case reports, series of fewer than 15 subjects, reports that combined fingernail onychomycosis and toenail onychomycosis in their statistics, and articles reporting only the total number of toenails cured without providing data as to how many subjects were cured. DATA EXTRACTION: This elimination process left 26 articles, which were then analyzed by means of a checklist that included a morphologically normal nail, mycological findings, and methodological items including recurrence rate, intent-to-treat analysis, placebo group, and whether terms were defined. DATA SYNTHESIS: When there was sufficient data, the frequency with which the treatment achieved normal mycological results, a clinically normal nail, and a disease-free nail (normal appearing nail plus normal mycological results) was calculated. Confidence intervals were calculated for disease-free nail results. CONCLUSIONS: Standard courses of terbinafine achieved a disease-free nail in approximately 35% to 50% of patients. For itraconazole, the relevant disease-free nail rate was about 25% to 40%. Disease reappearance is an important issue; unfortunately data are lacking as to its frequency. PMID- 9875193 TI - Does oral zinc aid the healing of chronic leg ulcers? A systematic literature review. AB - OBJECTIVE: To determine whether oral zinc sulfate is an effective treatment for promoting healing of venous or arterial leg ulcers. DATA SOURCES: The search strategy of the Cochrane Wounds Group was used. This includes searches of electronic databases, conference proceedings, relevant bibliographies, and hand searching of journals. STUDY SELECTION: Studies were included if they were randomized controlled trials of oral zinc sulfate in the treatment of chronic venous or arterial ulcers with objective measures of healing. Six of the 10 studies initially identified were included in the review. DATA EXTRACTION: The trial method, participants, interventions, outcomes, baseline comparability, adequate reporting of withdrawals, and blinding of assessment were extracted by 2 reviewers independently. DATA SYNTHESIS: No trial showed a statistically significant benefit of zinc sulfate for healing leg ulcers. There is limited evidence to suggest that zinc might increase healing in individuals with a low serum zinc level, but more evidence is needed. CONCLUSIONS: There is no evidence of benefit from the general use of zinc sulfate in patients with chronic leg ulcers. There is a need for further research to see if oral zinc sulfate is beneficial in the treatment of patients with leg ulcers who have a low serum zinc level. If it is demonstrated to be beneficial, further trials are required to establish dose and duration of treatment. PMID- 9875194 TI - Epiluminescence microscopy for the diagnosis of doubtful melanocytic skin lesions. Comparison of the ABCD rule of dermatoscopy and a new 7-point checklist based on pattern analysis. AB - OBJECTIVE: To compare the reliability of a new 7-point checklist based on simplified epiluminescence microscopy (ELM) pattern analysis with the ABCD rule of dermatoscopy and standard pattern analysis for the diagnosis of clinically doubtful melanocytic skin lesions. DESIGN: In a blind study, ELM images of 342 histologically proven melanocytic skin lesions were evaluated for the presence of 7 standard criteria that we called the "ELM 7-point checklist." For each lesion, "overall" and "ABCD scored" diagnoses were recorded. From a training set of 57 melanomas and 139 atypical nonmelanomas, odds ratios were calculated to create a simple diagnostic model based on identification of major and minor criteria for the "7-point scored" diagnosis. A test set of 60 melanomas and 86 atypical nonmelanomas was used for model validation and was then presented to 2 less experienced ELM observers, who recorded the ABCD and 7-point scored diagnoses. SETTINGS: University medical centers. PATIENTS: A sample of patients with excised melanocytic lesions. MAIN OUTCOME MEASURES: Sensitivity, specificity, and accuracy of the models for diagnosing melanoma. RESULTS: From the total combined sets, the 7-point checklist gave a sensitivity of 95% and a sepcificity of 75% compared with 85% sensitivity and 66% specificity using the ABCD rule and 91% sensitivity and 90% specificity using standard pattern analysis (overall ELM diagnosis). Compared with the ABCD rule, the 7-point method allowed less experienced observers to obtain higher diagnostic accuracy values. CONCLUSIONS: The ELM 7-point checklist provides a simplification of standard pattern analysis because of the low number of features to identify and the scoring diagnostic system. As with the ABCD rule, it can be easily learned and easily applied and has proven to be reliable in diagnosing melanoma. PMID- 9875195 TI - Efficacy and prognostic value of simple wound measurements. AB - OBJECTIVES: To assess the association between planimetric wound area and simple wound measurements. To ascertain the applicability of planimetry-based research to clinical practice. DESIGN: A retrospective cohort study of 260 consecutive patients followed up for at least 24 weeks. SETTING: Outpatient dermatology department of a major medical center. INTERVENTION: Wounds were measured using computer-based planimetry and were treated according to a standard protocol for venous leg ulcers. PATIENTS: Two hundred sixty patients with venous leg ulcers. MAIN OUTCOME MEASURES: Complete ulcer healing at 24 weeks. RESULTS: Using the Pearson, Spearman, and Kendall correlation coefficients, we found a tight correlation between planimetric wound area and wound width, length, width x length, perimeter, and area based on the formula for an ellipse. The values for all correlation coefficients were greater than 0.80 (P < .001 for all associations). Width, length, length x width, and elliptical area measurements all correlated closely with failure to heal (P < .001). The correlation between simple values and planimetric area dropped considerably for wounds more than 40 cm2, which represented 6% of the wounds in this study. CONCLUSIONS: The equivalence between simple and planimetric wound measurements allows for the extrapolation of wound measurements from clinical trials to clinical practice, and suggests that simple wound measurements may confidently be used as predictors of healing in the clinical setting. PMID- 9875196 TI - The effectiveness of a history-based diagnostic approach in chronic urticaria and angioedema. AB - OBJECTIVE: To assess the value of extensive laboratory screening for the identification of causes in patients with chronic urticaria and/or angioedema. DESIGN: In a prospective study involving 220 patients, 2 diagnostic strategies were compared: the combination of detailed history taking and limited laboratory investigations vs detailed history taking and extensive laboratory screening. The results of the extensive screening program were initially kept secret from the patients and the physicians. Later, all results were disclosed, and an investigation was undertaken to find out whether this information changed the initial diagnosis. The patients were followed up for 1 year to evaluate the results of interventions and to detect latent causes. SETTING: The study was performed in the outpatient department of a secondary and tertiary care center with institutional practice. PATIENTS: A total of 238 consecutive new patients with chronic urticaria and/or angioedema edema were referred; 18 of them refused participation. One patient was unavailable for follow-up. MAIN OUTCOME MEASURE: The difference in the number of identified causes between both approaches and the nature of the causes that would have been missed by omitting extensive laboratory screening. RESULTS: With a questionnaire and the limited laboratory tests, a cause was found in 45.9% of the patients, compared with 52.7% with the questionnaire and the extended screening program. Except for one parasitic infection, missed diagnoses were mainly adverse reactions to drugs or food detected by standard elimination procedures, not by laboratory investigations. CONCLUSION: Routine laboratory screening did not contribute substantially to the diagnosis of chronic urticaria or to the detection of underlying disorders. PMID- 9875197 TI - Risk of squamous cell carcinoma and methoxsalen (psoralen) and UV-A radiation (PUVA). A meta-analysis. AB - OBJECTIVE: To assess the risk of squamous cell carcinoma (SCC) and the relation of dose to risk among groups of patients with psoriasis exposed to psoralen-UV-A (PUVA). DATA SOURCES: Four electronic databases were searched from 1984 to 1998. STUDY SELECTION: In addition to the PUVA Follow-up Study, we included all English language studies from the United States and Europe with at least 150 patients enrolled, who were followed up for at least 5 years as identified from our bibliographic search. DATA EXTRACTION: A custom-designed questionnaire was used to extract data from each of the articles. For each study, if possible, we determined the incidence of basal cell carcinomas and SCCs and the incidence rate ratio of SCC among patients exposed to low-dose (we defined as < 100 treatments or 1000 J/cm2) compared with high-dose PUVA (> 200 treatments or 2000 J/cm2). Exact methods were used to calculate the incidence rate ratios. DATA SYNTHESIS: In addition to our study, we identified and reviewed 8 other studies. Overall, the incidence among patients exposed to high-dose PUVA was 14-fold higher than among patients with low-dose exposure (95% confidence interval, 8.3-24.1); a greater dose-dependent increase in risk than that observed in the PUVA Follow-up Study. CONCLUSION: Although the incidence of SCC reported among groups of PUVA treated patients followed up for at least 5 years varies greatly, compared with the risk in low-dose patients, long-term high-dose exposure to PUVA was consistently observed to significantly increase the risk of SCC in all studies reviewed. PMID- 9875198 TI - Comparison of the use of standardized diagnostic criteria and intuitive clinical diagnosis in the diagnosis of common viral warts (verrucae vulgaris). AB - OBJECTIVE: To assess the diagnostic value of standardized diagnostic criteria compared with the clinical intuitive diagnosis for verrucae vulgaris. DESIGN: A blind comparison with cross-over of experienced dermatologists using either intuitive clinical diagnosis or else recording the presence of standardized diagnostic criteria in verrucae vulgaris. SETTING: Clinical outpatient department in a major teaching hospital. PATIENTS: Fifteen patients aged 6 to 60 years with 21 verrucae vulgaris and 24 control lesions. INTERVENTION: Dermatologists examined the lesions and recorded either the diagnosis by the usual clinical intuitive method or else whether standardized diagnostic criteria were present in the lesion of each patient. Each dermatologist did 1 only of the 2 methods for each lesion with the other doing the alternative method for the same lesion, neither of them having previously been aware of any of the diagnoses. MAIN OUTCOME MEASURE: The sensitivity, specificity, and positive predictive, negative predictive, and relative values for each diagnostic approach were compared. RESULTS: Use of intuitive clinical diagnosis resulted in 100% specificity, sensitivity, and positive predictive, negative predictive, and relative values for both the verrucae vulgaris and the control lesions. Recording the presence of the diagnostic criteria reduced to a varying extent all the parameters of diagnostic accuracy with site--fingers and hands, elbows, and knees--being the most efficacious of the criteria. CONCLUSIONS: These data indicate that care is required in relying too much on standardized criteria as the basis for clinical diagnosis, even for a lesion as simple as a wart. They also reinforce the value of clinical experience obtained from repeated exposure to multiple variants of a disease in the traditional bedside teaching model. PMID- 9875199 TI - The development of practice guidelines for the treatment of severe plaque form psoriasis. AB - OBJECTIVE: To develop and introduce evidence-based guidelines for the selection of 5 commonly used treatment modalities--UV-B, photochemotherapy, methotrexate, acitretin, and cyclosporine--for adult patients with severe plaque form psoriasis. PATIENTS AND SETTING: Patients, residents, and dermatologists from the Department of Dermatology of the Academic Medical Center of the University of Amsterdam, Amsterdam, the Netherlands, were involved in this process. DESIGN: The development process started with a questionnaire to evaluate how patients with severe psoriasis were treated. A systematic literature review was set up to provide evidence-based estimates of effectiveness, adverse effects, and dropout rates. In 2 meetings, the opinion leaders and intended users discussed the results of the questionnaire and systematic review as well as the clinical considerations in the treatment choices. Guidelines were then made regarding the sequence of selection of 5 modalities in the concept of rotational therapy. These guidelines were introduced. Their use was analyzed for 6 months. RESULTS: Before the guidelines, there was no uniform approach. In the systematic review, 665 studies concerning the treatments were found. Exclusion rates were high. No studies of methotrexate therapy could be included. Photochemotherapy showed the highest average proportion of patients with clearance (70% [6947/9925]) and good response (83% [8238/9925]), followed by UV-B (67.9% [620/913]) and cyclosporine (64% [1030/1609]) therapy. In the second internal meeting, the following sequence for the treatments was defined: UV-B, photochemotherapy, methotrexate, acitretin, and cyclosporine. In 78% (69/88) of patients treated after the introduction, the guidelines were followed to determine the treatment choice. CONCLUSIONS: Guidelines for treating severe plaque form psoriasis can be successfully developed, introduced, and implemented and were considered to improve clinical care. PMID- 9875200 TI - Validation of a melanoma prognostic model. AB - BACKGROUND: A "clinically accessible," 4-variable (patient age, patient sex, tumor location, and tumour thickness) prognostic model has been published previously. This model evaluated variables that were commonly available to the clinician. Because models are heuristic, validity of a prognostic model should be evaluated in a population different from the original population. OBJECTIVE: To evaluate the external validity of this 4-variable melanoma prognostic model. DESIGN: To estimate the external validity of this model, we used a population based cohort of individuals with melanoma. We also evaluated a 1-variable model (tumor thickness). Estimates of the external validity of these logistic regression models were made using the c statistic and the Brier score. SETTINGS AND PATIENTS: A total of 1261 patients with melanoma evaluated in a multispecialty, university-based practice and 650 patients with melanoma from throughout Connecticut. MAIN OUTCOME MEASURE: Death from melanoma within 5 years of diagnosis. RESULTS: The c statistics for the 4-variable model were 0.86 (95% confidence interval [CI], 0.83-0.89) for the university-based practice data set and 0.81 (95% CI, 0.75-0.86) for the Connecticut data set. For thickness alone, the c statistics were 0.83 (95% CI, 0.80-0.86) and 0.79 (95% CI, 0.74-0.85), respectively. Brier scores for the 4-variable model were 0.09 (95% CI, 0.08-0.10) and 0.08 (95% CI, 0.06-0.09) and for the 1-variable model were 0.09 (95% CI, 0.08 0.10) and 0.08 (95% CI, 0.07-0.10), respectively. No significant differences exist between the data sets for the 4- and 1-variable models. CONCLUSIONS: The 4- and 1-variable models are generalizable. The simpler 1-variable model--tumor thickness--can be used with a relatively small loss in accuracy. PMID- 9875201 TI - Cost-effectiveness and cost-benefit analysis of using methotrexate vs Goeckerman therapy for psoriasis. A pilot study. AB - OBJECTIVE: To analyze the net benefit and cost-effectiveness of methotrexate use and Goeckerman therapy for psoriasis. DESIGN: Net benefit and cost-effectiveness depend on the costs, efficacy, and utilities of therapy. Utilities are quantitative measures of patient preferences. We obtained costs by using resource based accounting techniques. Efficacy was estimated from literature reports. We surveyed patients with psoriasis, dermatologists, and healthy subjects using utility assessment methods. All assumptions were examined in a sensitivity analysis. MAIN OUTCOME MEASURES: For net benefit, if benefits out-weighed the costs, it was deemed worth providing. For the cost-effectiveness analysis, the ratio of costs-to-effectiveness of less than $35,000 was considered cost effective. RESULTS: Using utilities from healthy nonexperts, the costs of both therapies exceeded the benefits in mild and moderate psoriasis. In severe psoriasis, only methotrexate demonstrates a net benefit. Both therapies were cost effective compared with no therapy. Liquid methotrexate should be chosen over the tablet form since it was cheaper and had the same outcome. Goeckerman was cost effective against liquid methotrexate in severe, but not mild or moderate psoriasis. There was a trend for therapies to be more cost-effective when using patient utilities and less with dermatologist utilities. The results were highly sensitive to efficacy and utilities. CONCLUSIONS: The results of this study need to be confirmed in other settings, but they demonstrate that the tools of cost effectiveness and cost-benefit analysis have great potential value in dermatology. Once efficacy is better characterized and utilities better quantified, these types of analyses will be crucial for health care policy. PMID- 9875202 TI - Evidence-based medicine in a nutshell. A guide to finding and using the best evidence in caring for patients. AB - Evidence-based medicine is the use of the best current evidence in making decisions about the care of individual patients. Practicing EBM requires recognition that in most encounters with patients, questions arise that should be answered to provide the patient with the best available medical care. Asking well built clinical questions that contain 4 elements--a patient or problem, an intervention, a comparison intervention (if necessary), and an outcome--is an important step in practicing EMB. Once appropriate questions have been formulated, the best source for finding most types of best evidence is by searching the MEDLINE database by computer. MEDLINE searches have inherent software and operator limitations that make their reliability quite variable. One should be aware of these limitations and improve one's skills in searching. The Cochrane Collaboration Controlled Clinical Trials Registry contains more than 190,000 controlled clinical trials and is the best source of evidence about treatment. The quality (strength) of evidence is based on a hierarchy of evidence: results of systematic reviews of well-designed clinical studies, results of 1 or more well-designed clinical studies, results of large case series, expert opinion, and personal experience. Once the best evidence has been found, the EBM approach involves critically appraising the quality of the evidence, determining its magnitude and precision, and applying it to the specific patient. Guidelines to critically appraise and apply evidence are available. The clinical question, best evidence, and its critical appraisal should be saved in a format that can be easily retrieved for future use. PMID- 9875203 TI - The Cochrane Skin Group. Preparing, maintaining, and disseminating systematic reviews of clinical interventions in dermatology. AB - In 1979, Prof Archie Cochrane challenged the medical profession to produce a critical summary of randomized controlled clinical trials according to specialty, which should be updated periodically. The Cochrane Collaboration, an international voluntary group of reviewers and researchers from a range of professional backgrounds dedicated to producing systematic reviews, was established in 1992 in response to Cochrane's challenge. Systematic reviews produced by the Cochrane Collaboration start with individuals who formulate questions that are important to the care of patients. Every effort is then made to locate published and unpublished evidence to answer the question, and explicit criteria are used to select studies for inclusion in the review and to assess their quality. If appropriate, meta-analysis is used to combine results from several smaller studies to produce an overall result. Reviews are published in the Cochrane Library, an electronic publication (CD-ROM or diskette form), and the reviews are updated quarterly. In December 1997, a Cochrane Skin Group was registered with the Cochrane Collaboration to prepare, maintain, and disseminate reviews on the effects of health care for people with dermatological conditions. Currently, 25 titles and 9 review protocols have been registered with the Cochrane Skin Group, and the first set of dermatological reviews will be available before the end of 1999. The Cochrane Skin Group aims to become the best source of unbiased external evidence for summarizing the effects of dermatological care. PMID- 9875204 TI - Asymptomatic swelling of a man's ear. Auricular pseudocyst. PMID- 9875205 TI - Multiple translucent facial papules. Apocrine hidrocystoma. PMID- 9875206 TI - Enlarging facial nodule on an elderly patient. Lymphoepithelioma-like carcinoma of the skin (LELCS). PMID- 9875207 TI - Dome-shaped partly umbilicated tumor on the ear. Spitz nevus (SN). PMID- 9875208 TI - PURPLE (oops! Atrophie blanche) revisited. PMID- 9875209 TI - Lack of association between aphthous ulcers and Helicobacter pylori. PMID- 9875210 TI - PAAn-1b and PAAn-E: two phosphorothioate antisense oligodeoxynucleotides inhibit human aromatase gene expression. AB - Estrogen-dependent diseases, especially breast cancers, are frequently treated with aromatase inhibitors. Another more recent strategy is the antisense technology. In this study, after predicting aromatase mRNA secondary structure, we describe the design, the efficiency, and the toxicity of two antisense phosphorothioate oligodeoxynucleotides (PAAn-1b and PAAn-E) directed toward aromatase mRNA. Indeed, 2 microM PAAn-1b and PAAn-E encapsulated with 54 microM polyethylenimine inhibit aromatase activity by 71 and 79%, respectively, in transfected 293 cells, with IC50 values of 0.2 and 0.6 microM. The mechanism of inhibition appears to be specific after using sense and scramble oligodeoxynucleotides as controls and largely decreases aromatase mRNA and protein amounts. Moreover, PAAn-1b and PAAn-E are not cytotoxic for 293 cells. This study finally provides a new strategy for aromatase inhibition. It offers new tools for studying aromatase gene expression and its role in cancer for instance, and this could be of help for the therapy of estrogen-dependent diseases. PMID- 9875211 TI - Cloning and tissue distribution of novel splice variants of the rat GABAB receptor. AB - We have identified two novel splice variants of the metabotropic gamma aminobutyric acid receptor (GABABR1), designated GABABR1c and GABABR1d, when screening a rat cerebellum cDNA library. GABABR1c has an amino acid sequence identical to GABABR1b, a member of GABABR1 isoforms, and an additional 93-bp insertion that generates an additional 31-amino-acid sequence in the fifth transmembrane region of GABABR1b. Thus, GABABR1c may have a structural variation in the second extracellular loop and fifth transmembrane region. GABABR1d also has an amino acid sequence identical to GABABR1b and an additional insertion of 566 bp that generates a divergent amino acid sequence in the carboxylterminal end. Reverse-transcription polymerase chain reaction analysis showed that in various rat tissues GABABR1c mRNA was ubiquitously expressed and GABABR1d mRNA in forebrain, cerebellum, eye, kidney, and urinary bladder. GABABR1 isoforms may function not only in the central nervous system but also in various peripheral tissues. PMID- 9875212 TI - Polar amino acid-rich sequences bind to polyglutamine tracts. AB - Polyglutamine tracts are found in different proteins including transcription factors and cofactors as well as in triplet repeat disease gene products. To characterize the protein motif that binds to the polyglutamine tract, we screened a human embryonic brain cDNA library with the polyglutamine tract of Brn-2 as bait using the yeast two-hybrid method. All six isolated clones encoding polyglutamine tract binding proteins were rich in polar amino acids. Three of these clones could form polar helical structures. These observations suggest that polar amino acid-rich sequences are essential for binding to the polyglutamine tract. PMID- 9875213 TI - Rho-associated kinase phosphorylates desmin, the myogenic intermediate filament protein, at unique amino-terminal sites. AB - We obtained evidence that Rho-associated kinase (Rho-kinase) phosphorylates desmin, the myogenic intermediate filament protein, with approximately 2 mol phosphate per mole of desmin in vitro. Desmin phosphorylated by Rho-kinase lost the potential to form 10-nm filaments. Thr-16, Thr-75, and Thr-76 on desmin proved to be the major phosphorylation sites for Rho-kinase. All these sites are located within the head domain and are different from the reported phosphorylation sites of protein kinase. A, protein kinase C, and cdc2 kinase. We are entertaining the notion that Rho-kinase may regulate filament structures of desmin by site-specific phosphorylation. PMID- 9875214 TI - Erythropoietin prevents place navigation disability and cortical infarction in rats with permanent occlusion of the middle cerebral artery. AB - Erythropoietin (EPO) prevents the ischemia-induced delayed neuronal death in the hippocampal CA1 field in gerbils. EPO receptor (EPOR) is also expressed in the cerebral cortex but its function is not known. To examine whether EPO has a neuroprotective action in the cortex, EPO was infused into the cerebroventricles of stroke-prone spontaneously hypertensive rats with permanent occlusion of the left middle cerebral artery. Morris water maze test indicated that EPO infusion alleviated the ischemia-induced place navigation disability. The left (ischemic) to-right (contralateral nonischemic) (L/R) ratio of cerebrocortical area in the EPO-infused ischemic group was larger than that in the vehicle-infused ischemic group. The occlusion caused secondary thalamic degeneration but infusion of EPO prevented the decrease in the L/R ratio of thalamic area and supported neuron survival in the ventroposterior thalamic nucleus. In situ hybridization indicated that EPOR mRNA was upregulated in the periphery (ischemic penumbra) of a cerebrocortical infarct after occlusion of the middle cerebral artery, suggesting that an increased number of EPOR in neurons facilitates the EPO signal transmission, thereby preventing the damaged area from enlarging. PMID- 9875215 TI - MAPKKK6, a novel mitogen-activated protein kinase kinase kinase, that associates with MAPKKK5. AB - MAPKKK5/ASK1 activates c-Jun N-terminal kinase (JNK) and p38 kinase signaling pathways and induces apoptosis when expressed in stably transfected cells. Using MAPKKK5 as bait in yeast two-hybrid screening, a novel protein that interacts with MAPKKK5 was identified and cloned. This novel protein is predicted to contain all 11 kinase subdomains and shares 45% amino acid identity with MAPKKK5 and thus is designated MAPKKK6. The interaction of MAPKKK6 with MAPKKK5 in vivo was confirmed by coexpression of MAPKKK5 and MAPKKK6 in 293 cells followed by immunoprecipitation. In contrast to MAPKKK5, which activated both JNK and p38 kinase pathways, MAPKKK6 only weakly activated JNK but not ERK or p38 kinase pathways. PMID- 9875216 TI - Genistein-mediated attenuation of tamoxifen-induced antagonism from estrogen receptor-regulated genes. AB - In this study we demonstrate that physiologic concentrations of genistein are sufficient to mediate agonism and to reverse the repressive effects of 4 hydroxytamoxifen on estrogen receptor (ER alpha)-responsive reporter genes. We also show that overexpression of the steroid receptor coactivator (SRC-1) potentiates transactivation by genistein-activated ER alpha and that coexpression of CBP (the cAMP response element binding protein coactivator) synergistically increases this signal. Exogenous expression of a nuclear receptor corepressor (NCoR) was, however, unable to alter genistein-mediated transactivation. In in vitro binding assays, we show that genistein, but not 4-hydroxytamoxifen, induces a direct interaction between radiolabeled ER alpha and a GST-SRC-1 fusion protein. More importantly, coincubation with genistein and 4-hydroxytamoxifen or genistein treatment following preincubation of the ER with 4-hydroxytamoxifen also resulted in a strong physical interaction with SRC-1. These findings imply that genistein-induced shifts in the coregulator status of ER alpha may be involved in transcriptional regulation and suggest that tamoxifen-mediated antagonism at ER-dependent genes is sensitive to attenuation by low levels of genistein. PMID- 9875217 TI - Dioxin causes a sustained oxidative stress response in the mouse. AB - Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin; TCDD) is the prototype for environmental agonists of the aromatic hydrocarbon receptor (AHR) that are known to produce multiple adverse effects in laboratory animals as well as humans. Although not directly genotoxic, dioxin is known to increase transformation and mutations in mammalian cell culture and to cause an exaggerated oxidative stress response in the female rat. In humans and mice, however, dioxin-mediated oxidative stress appears to be more subtle, causing a response that has been poorly characterized. Using the female C57BL/6J inbred mouse, we show here that intraperitoneal treatment of 5 micrograms TCDD per kilogram on 3 consecutive days produces a striking, prolonged oxidative stress response: hepatic oxidized glutathione levels increase 2-fold within 1 week, and these effects persist for at least 8 weeks despite no further dioxin treatment. Urinary levels of 8 hydroxydeoxyguanosine--a product of DNA base oxidation and subsequent excision repair--remain elevated about 20-fold at 8 weeks after dioxin treatment, consistent with chronic and potentially promutagenic DNA base damage. These results demonstrate that dioxin exposure does produce a sustained oxidative stress response in the mouse. PMID- 9875218 TI - Presence and formation of heme and occurrence of certain heme proteins in the filarial parasite Setaria digitata. AB - There is currently renewed interest in the biological significance of heme proteins. The most common heme proteins include hemoglobin, myoglobin, cytochromes, and redox enzymes such as catalase and peroxidase. Setaria digitata is a cattle filarial parasite, which is devoid of typical cytochrome systems. However, studies showed activities of delta Aminolevulinate synthase (ALAS), delta Aminolevulinate dehydratase (ALAD), and heme oxygenase in appreciable amounts, suggesting the presence of necessary equipment for the biosynthesis of heme. This is further confirmed by the end product inhibition of ALAS by heme and the observation of the death of the parasite by succinyl acetone, an inhibitor of the biosynthesis of heme. Though typical cytochrome systems are absent, microsomal cytochrome P 450 and elevated levels of heme containing enzymes such as catalase and peroxidase are present in the parasite. A unique hemoglobin is also detected which shows a difference in biological functions from the host system and that of the much-studied nematode parasite Ascaris sum. PMID- 9875219 TI - Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi. AB - Cruzipain, the major proteinase of Trypanosoma cruzi, plays an important role in the biology of this parasite. This study reports the development of a recombinant Fab antibody, using RNA isolated from the anti-Ag163B6 hybridoma against cruzipain. This procedure involves the use of cDNAs obtained with the aid of a specific set of primers complementary to the complete light kappa chain (L kappa) and the first two domains of the IgG1 heavy chain (VH/CH1). These products were subsequently cloned in the pComb3 system, from which the gIII gene had been removed, and expressed in Escherichia coli cells. The recombinant Fab molecule recognized cruzipain by ELISA, in a fashion similar to the original mAb anti Ag163B6. Nucleotide sequence analysis of the recombinant molecule, together with its immunological recognition by specific anti-mouse IgG (Fab)2, indicated the immunoglobulin nature of the recombinant product. Moreover, both the mAb anti Ag163B6 and the soluble Fab fragment described here react similarly with the intact parasite surface, as observed in an indirect immunofluorescence assay. In conclusion, our recombinant Fab anti-Ag 163B6 allows the possible use of this molecule for diagnosis, antigen purification, and eventually treatment of Chagas afflicted individuals. PMID- 9875220 TI - Acute infection of Sindbis virus induces phosphorylation and intracellular translocation of small heat shock protein HSP27 and activation of p38 MAP kinase signaling pathway. AB - In general, viral infection is supposed to induce stress responses in the host cell. However, very few detailed observations about virus-induced stress responses have been reported. Here we investigated specific stress responses in Vero cells infected with Sindbis virus (SV), a single-stranded RNA virus, acute infection with which is known to cause apoptotic cell death in the host cells. Prior to the onset of apoptosis, p38 mitogen-activated protein kinase (MAPK) and c-Jun NH2-terminal kinases (JNKs) were activated. Subsequently, a 27-kDa heat shock protein (HSP27) became phosphorylated, and intracellular distribution of HSP27 was changed from the cytoplasm to the perinuclear region. These results indicate that the cellular signaling cascades activated by pro-inflammatory cytokines and environmental stresses are also activated as a result of lytic infection with SV. These responses may contribute to the delayed onset of apoptosis in the host cells and the facilitation of viral replication. PMID- 9875221 TI - A mouse model for beta cell-specific ablation of target gene(s) using the Cre loxP system. AB - The rat insulin promoter (RIP) has been used to drive the expression of Cre recombinase (Cre) specifically in beta cells. Transient transfection was performed in the mouse insulinoma cell line, NIT-1, and control cell lines. RT PCR was performed using total RNA from pancreas and other tissues of RIP-Cre transgenic mice. In addition, the efficiency and specificity of RIP were further analyzed by cross breeding the RIP-Cre transgenic mice with reporter mice bearing a beta-actin-loxP-CAT-loxP-lacZ transgene. In these mice, lacZ is expressed only after excision of the floxed-CAT gene by Cre-mediated recombination. Here, we present the data for beta cell-specific expression of lacZ in bigenic mice, as proof of concept in a mouse model for targeting beta cell-specific gene(s). The RIP-Cre transgenic mice will be used as a potential tool for targeting the excision of beta cell-specific gene(s) to study their role in islet cell physiology. PMID- 9875222 TI - Comparison of predicted and observed properties of proteins encoded in the genome of Mycobacterium tuberculosis H37Rv. AB - Proteome studies complement current molecular approaches through analysis of the actively translated portion of the genome (the "functional proteome"). Two dimensional gel electrophoresis (2-DGE) utilising immobilized pH gradients of pH 2.3-5.0 and pH 6.0-11.0, developed with predetermined regions of overlap compatible with commercially available pH 4.0-7.0 gradients, permitted the display of a significant portion of the proteome of Mycobacterium tuberculosis H37Rv. A significant portion of the M. tuberculosis proteome, in the molecular mass (M(r)) window 5 kDa to 200 kDa and with isoelectric point (pI) between pH 2.3 and 11.0, was visualised for the first time. A total of 493 protein spots were effectively resolved, including 126 spots that could not be seen using standard pH 4.0-7.0 gradients. These results were used to compare the physical properties of the observed proteins to the theoretical predictions of the recently completed M. tuberculosis H37Rv genome. Most proteins were found in the pI and mass window of pH 4.0-7.0 and 10-100 kDa. Analysis of the predicted proteome revealed a bimodal pI distribution, with substantial numbers of proteins in the pI regions 4.0-7.0 and 9.0-12.0 as has been seen for the majority of completed genomes. Such data may reveal current limitations in experimental extraction and separation of extremely basic, high M(r) and hydrophobic proteins via 2-DGE. Conversely, 13 acidic proteins were observed with pI less than the lowest value predicted by the genome. In addition, a subset of small protein (< 10 kDa) were observed within the pI region of pH 5.0-8.0 that were not predicted by the complete genomic sequence, reflecting the current inability to distinguish small genes from within DNA sequence. This work represents the foundation for comparing the protein expression patterns of different pathogenic and nonpathogenic M. tuberculosis strains. The characterization of M. tuberculosis protein expression, further facilitated by the recent completion of the genome sequence, could aid in developing more effective diagnostic or therapeutic reagents. PMID- 9875223 TI - Expression of spliceosome-associated protein 49 is required for early embryogenesis in Caenorhabditis elegans. AB - Spliceosome-associated protein 49 (SAP 49) is a subunit of the splicing factor SF3b, which is involved in 3' splice site recognition in pre-mRNA splicing. Here we show the Caenorhabditis elegans SAP 49 gene is located in a gene cluster that is transcribed polycistronically with three other upstream genes. Transgenic analysis of expression of the gene cluster under the control of its native promoter revealed that in spite of its predicted essential function for all cells, the C. elegans SAP 49 expression is limited to specific cells in both larval and adult stages. When the endogenous SAP 49 expression was inhibited by microinjection of an antisense RNA, embryos laid by the injected worms ceased to develop at a specific stage of early embryogenesis, indicating that SAP 49 plays an essential role in the development of C. elegans. These results raise the possibility that SAP 49 is a cell-specific, not constitutive, splicing factor. PMID- 9875224 TI - Differential regulation of mouse uncoupling proteins among brown adipose tissue, white adipose tissue, and skeletal muscle in chronic beta 3 adrenergic receptor agonist treatment. AB - Uncoupling proteins (UCPs) are inner mitochondrial membrane transporters that dissipate the proton gradient, releasing stored energy as heat, without coupling to other energy-consuming processes. Therefore, the UCPs are thought to be important determinants of the metabolic efficiency. To elucidate relationships between the UCPs expressions and insulin sensitivity improvement, we treated KK Ay mice with beta 3 adrenergic receptor agonist for 21 days and examined the changes of the UCPs mRNA expressions in various tissues. Chronic treatment of a specific beta 3 adrenergic receptor agonist, CL316,243 (0.2 mg/kg body weight/day s.c.) markedly increased the expressions of uncoupling protein 1 (UCP1), uncoupling protein 2 (UCP2), and uncoupling protein 3 (UCP3) by 14-fold, 6-fold, and 16-fold, respectively, in the brown adipose tissue (BAT). The UCP1 and UCP3 mRNA expressions in the white adipose tissue (WAT) were also increased by 12-fold and 9-fold, respectively, but the UCP2 mRNA expression was not changed in this tissue. Interestingly, the UCP2 and UCP3 mRNA expressions were strikingly decreased in the skeletal muscle and heart. Particularly, the UCP3 mRNA expression level in the skeletal muscle was dropped to 10% of that of the saline treated control mice, indicating that the UCPs mRNA expressions are regulated in tissue-specific ways. The concentrations of plasma insulin and circulating free fatty acid (FFA) were significantly decreased, suggesting that they correlate with the reductions of the UCP2 and UCP3 mRNA expressions in the skeletal muscle and heart. It has been thought that the UCP1 and UCP3 mRNA expressions in the BAT and WAT are mainly controlled by the hypothalamus via the sympathetic nervous system, while the levels of insulin, FFA or both may play important roles in the control of the UCP2 and UCP3 mRNA expressions in the skeletal muscle an heart. PMID- 9875226 TI - Spin trapping of nitric oxide in aqueous solutions of cigarette smoke. AB - Nitric oxide, a gaseous free NO radical (.NO) generated in particulate-free gas phase main-stream smoke of cigarettes, was observed with electrical spin resonance (ESR) using a spin trapping technique. N-Methyl-D-glucamine dithiocarbamate (MGD)2-Fe2+ complex was used for the NO radical spin trapper in aqueous solution. The intensity of the ESR signal of the spin adduct formed by bubbling smoke from one cigarette increased gradually with time over 2 hours at about 20 degrees C and was constant for 2 days or longer. The time course of the production of the NO radical followed the rate equation y = 1520(1-e-0.018t) for the first-order reaction up to around 25 min after mixing of Fe2+ solution and then slowly approached the maximum value determined by the concentration of the spin adduct. These findings suggest that NO radical is produced slowly from NO radical donors such as amine .NO complexes, peroxinitrite (ONOO-), and other reactants such as nitrogen oxides (NOx), which are produced from the smoke of tobacco leaves, and suggest that its generation could be involved in the decomposition or cleavage of such substances. PMID- 9875225 TI - Cleavage of human cytosolic phospholipase A2 by caspase-1 (ICE) and caspase-8 (FLICE). AB - The activation of caspases appears to play a key role in programmed cell death. An increasing number of substrates have been identified that are cleaved by caspases. In a previous study, we have reported that human cPLA2 is proteolytically inactivated during apoptosis through cleavage by a caspase-3-like activity. Here, we show that in cotransfection experiments the previously identified cleavage site at Asp522 can be used by a wide variety of caspases belonging to different subfamilies. The formation of additional fragments implied differences in cleavage site usage between the closely related caspases-3 and -7. A different cleavage pattern of cPLA2 was observed with caspase-1. Mutational analysis identified the caspase-1 cleavage site at Asp459 within the sequence YQSD/N. Most interestingly, we found that even caspase-8, an upstream component of the proposed caspase cascade, cleaves cPLA2 in vitro. The presence of multiple cleavage sites warrants proteolysis and inactivation of the proinflammatory cPLA2 during apoptosis. PMID- 9875227 TI - Glutathione downregulates the phosphorylation of I kappa B: autoloop regulation of the NF-kappa B-mediated expression of NF-kappa B subunits by TNF-alpha in mouse vascular endothelial cells. AB - Nuclear factor-kappa B (NF-kappa B) regulates gene expression upon immune and inflammatory responses. It has been demonstrated that redox regulation by thiols is involved in the signal-transduction cascade. In this study, we examined the effect of glutathione (GSH) on the NF-kappa B activity and the expression of NF kappa B subunits induced by tumor necrosis factor-alpha (TNF-alpha) using mouse vascular endothelial cells. GSH inhibited the serine phosphorylation of I kappa B alpha by TNF-alpha, leading to the downregulation of NF-kappa B-DNA binding activity followed by decreased expression of p65/p50 and I kappa B mRNAs. The regulation of the autoregulatory loop for the NF-kappa B activation and the expression of NF-kappa B subunits may be important in endothelial cells in response to cytokines. PMID- 9875228 TI - Basic transcription element binding protein (BTEB) transactivates the cholesterol 7 alpha-hydroxylase gene (CYP7A). AB - Cholesterol 7 alpha-hydroxylase catalyzes the first and rate-limiting step in the conversion of cholesterol to bile acids in the liver. Previously, we have identified two bile acid response elements located in nt -74 to -54 (BARE-I) and 148 to -118 (BARE-II) regions. The nucleotide sequences in these BAREs are highly conserved and shared a novel sequence, AGTTCAAG. To identify and isolate nuclear protein factors that bind to these BAREs, we have screened a human liver cDNA expression library with oligonucleotide probes containing the sequence from nt 149 to -127. Twenty positive clones were selected and purified. Partial nucleotide sequences of these clones were determined. Nucleotide homology search of DNA databases of the sequences of these clones revealed that sequence of one clone, G13, is identical to basic transcription element binding protein (BTEB), a GC box-binding protein of Sp1 family transcription factors known to regulate many cytochrome P450 genes. Electrophoretic mobility shift assays have identified a basic transcription element (BTE) in BARE-II and a Sp1 binding site located in the nt -100/-82 region of the CYP7A promoter. Transient transfection assays have confirmed that BTEB was able to transactivate the CYP7A promoter/luciferase chimergic gene. PMID- 9875229 TI - Vitamin E and lipoic acid, but not vitamin C improve blood oxygenation after high energy IMPULSE noise (BLAST) exposure. AB - Exposure to high energy impulse noise (BLAST) caused by explosions, result in structural and functional damage to the hollow organs, especially to the respiratory and auditory systems. Lung damage includes alveolar wall rupture, edema and hemorrhage, and may be fatal. Previous observations at the molecular level using the rat model, suggested that secondary free radical-mediated oxidative stress occurs post exposure resulting in antioxidant depletion and hemoglobin (Hb) oxidation. This study examined whether a short period of pre exposure supplementation with antioxidants would protect Hb from the effects of BLAST exposure. Six groups of male Sprague-Dawley rats (8/group) were gavaged with 800 IU vitamin E (VE) in 2 ml corn oil, 1000 mg vitamin C (VC) in 2 ml distilled water or 25 mg or (-lipoic acid (LA) in 2 ml corn oil for 3 days. Matched control groups were gavaged with the respective vehicles. On day 4, rats were deeply anesthetized and exposed to a simulated BLAST wave with an average peak pressure of 62 +/- 2 kPa. Rats were euthanized one hour post exposure and blood samples were obtained by cardiac puncture and analyzed using a hemoximeter. Post exposure oxygenation states (HbO2, O2 saturation, and O2 content) were markedly decreased, while reduced-Hb was increased. Supplementation with VE and LA reversed the trend and increased Hb oxygenation, but VC did not. This suggests that a brief dietary loading with pharmacological doses of VE or LA, but not VC shortly before BLAST exposure may be beneficial. Moreover, measurement of blood oxygenation may function as a simple semi-invasive biomarker of BLAST-induced injury applicable to humans. PMID- 9875230 TI - Critical aggregation concentrations of gram-negative bacterial lipopolysaccharides (LPS). AB - Bacterial endotoxins or lipopolysaccharides (LPS), cell wall components of gram negative bacteria, are involved in septic shock. While the carbohydrate structure of LPS have been studied in the past, little is known about the macromolecular structure and formation of LPS fragments in blood. It is believed that amphiphilic molecules such as LPS occur as monomers and aggregate into macromolecular structures above a critical micelle or critical aggregate concentration, CAC. The CAC of Lipid A, a LPS precursor, and several LPS serotypes of varying molecular weight and different polysaccharide chain lengths were established by static light scattering and by steady-state fluorescence spectroscopy by incorporation of the fluorescent probe, NPN, N-phenyl-1 naphthylamine. The CAC for short polysaccharide chain mutant LPS S.minnesota Re 595 (MW 2,500) and Lipid A from S.minnesota Re 595 (MW 2000) were 4 microM and 5 microM respectively. The CAC of LPS from heterogeneous long O-antigen polysaccharide chain bacterial serotypes: S.minnesota wildtype were 11 micrograms LPS/ml, S.typhimurium 14 micrograms LPS/ml and E.coli 0111:B4 22 micrograms LPS/ml, respectively. The results obtained suggests that critical aggregate concentration and solubility of LPS is a function of polysaccharide chain length. PMID- 9875231 TI - Characterization of IgE and IgG epitopes on ovomucoid using egg-white-allergic patients' sera. AB - To investigate the importance of linear and conformational structure and carbohydrate chains in hen ovomucoid epitopes, the IgG and IgE binding activities of three native and reduced carboxymethylated (RCM) domains (DI, DII, and DIII) were compared using human sera from egg-white-allergic patients. There was significantly more IgG and IgE binding activity to DIII than to DI and DII. The IgG binding activity to RCM domains was similar to the native form, while RCM DIII had significantly greater binding activity to IgE antibody (p < 0.05). It indicated that the main IgE and IgG epitopes on each domain were of linear structure. However, the total reactivity of the three domains was estimated to be about 50-60% (IgG binding) and 55-75% (IgE binding) compared with total reactivity in ovomucoid, resulting in some ovomucoid epitopes consisting of conformational epitopes on domain I-II or II-III. The carbohydrate moieties in DIII had a rather inhibiting effect on its IgG and IgE binding activities. PMID- 9875232 TI - Plasmin, substilisin-like endoproteases, tissue plasminogen activator, and urokinase plasminogen activator are involved in activation of latent TGF-beta 1 in human seminal plasma. AB - We reported previously that TGF-beta 1 is a major immunosuppressive agent in human seminal plasma. TGF-beta 1 in seminal plasma is so abundant that it may represent the highest physiologic concentration of TGF-beta 1 reported for a biological fluid. The in vitro activation of TGF-beta 1 is detected at acidic pH. The acidic environment of the vagina is suggested as an in vivo physiological condition for the activation of seminal plasma latent TGF-beta 1. The present study demonstrates that Pefabloc [4-(2-aminoethyl)-benzenesulfonyl fluoride AEBSF]-inhibitable serine proteases are involved in the activation of latent TGF beta 1. Pefabloc inhibits latent TGF-beta 1 activation in a dose- and time dependent manner. The use of other protease inhibitors and specific antibodies reveals that, in addition to plasmin, substilisin-like endoproteases and tissue- and urokinase-type plasminogen activators participate in the activation of latent TGF-beta 1 in human seminal plasma. PMID- 9875233 TI - Purification and characterization of novel ribosome inactivating proteins, alpha- and beta-pisavins, from seeds of the garden pea Pisum sativum. AB - Two ribosome inactivating proteins designated alpha- and beta-pisavins were isolated from seeds of the garden pea Pisum sativum var. arvense Poir with a procedure involving affinity chromatography on Affi-gel Blue gel, immobilized metal ion affinity chromatography on Iminodiacetic acid-agarose, cation exchange chromatography on Resource-S, and gel filtration on Superose 12. alpha- and beta pisavins are nonglycoproteins with a molecular weight of 20.5 kDa and 18.7 kDa respectively. The sequences of the first sixty N-terminal amino acids of alpha- and beta-pisavins were identical. In isoelectric focusing these two proteins merged into one band with a pI greater than 9.3. Inhibition of protein synthesis by a rabbit reticulocyte lysate system was achieved at an IC50 of approximately 0.5 nM. Activity of the proteins toward tRNA was observed. The proteins acted on ribosomal RNA through its RNA N-glycosidase activity to release an Endo's fragment, and converted the conformation of DNA from supercoiled and circular forms into a linear form. PMID- 9875234 TI - Ribosome inactivating protein and lectin from bitter melon (Momordica charantia) seeds: sequence comparison with related proteins. AB - One of the ribosome inactivating proteins (RIPs), beta-momorcharin, and a lectin were isolated from seeds of the bitter melon Momordica charantia (Family Cucurbitaceae) in accordance with published procedures. Both beta-momorcharin and M. charantia lectin were then subjected to amino acid sequencing. beta momorcharin exhibited considerable homology in sequence to other Cucurbitaceae RIPs, and also to the A chains of abrin and ricin which are type II (double chained) RIPs. The resemblance between alpha-momorcharin and other RIPs is closer than that between beta-momorcharins and other RIPs. M. charantia lectin manifested a certain extent of sequence similarity to beta-momorcharin and other RIPs, and some degree of homology in sequence to lectins from Cucurbita maxima, Cucurbita argyrosperma, Sambucus nigra and Ricinus Communis. PMID- 9875235 TI - Angiotensin II interferes with leukemia inhibitory factor-induced STAT3 activation in cardiac myocytes. AB - Recently, we reported that leukemia inhibitory factor (LIF), a member of the interleukin (IL)-6 cytokine family, transduced hypertrophic and cytoprotective signals via Januas Kinase-signal transducer and activator of transcription (JAK STAT) pathway in cardiac myocytes. Angiotensin II (AII) is also known to activate STATs and reported to induced apoptosis in adult rat ventricular myocytes. In the present study, we investigated potential interactions between gp130 dependent and AII signaling pathways, by examining AII regulation of LIF-induced anti-apoptotic effect and STAT3 activation in cardiac myocytes. Although LIF attenuated the DNA fragmentation induced by serum depletion, AII augmented the DNA fragmentation in cultured neonatal rat cardiac myocytes. Furthermore, LIF-mediated cytoprotective effect was inhibited by AII pretreatment. LIF rapidly and transiently tyrosine phosphorylated STAT3 in cardiac myocytes which was not observed by AII. AII pretreatment inhibited LIF-induced phosphorylation of STAT3 in a dose dependent manner. This inhibitory effect of AII on STAT3 activation was blocked by the AII type I (AT1) receptor antagonist CV11974. These results demonstrate that negative crosstalk between gp130 and AT1 receptor dependent signaling exists in cardiac myocytes. This crosstalk may contribute to the modulation of pathophysiological process in myocardial disease. PMID- 9875236 TI - Inhibition of apoptosis-associated DNA fragmentation activity in nonapoptotic cells: the role of DNA fragmentation factor-45 (DFF45/ICAD). AB - We have investigated the mechanism whereby nuclear DNA fragmentation activity emerging during early apoptosis is inhibited during normal cell life. In a cell free system, cytosol fractions from diverse nonapoptotic human cell lines (Jurkat T-cell leukemia, HeLa carcinoma, SK-N-MC neuroblastoma, and WI-38 embryonic lung fibroblast) potently neutralized the nuclear DNA fragmentation activity of cytosol from apoptotic anti-Fas treated Jurkat cells. Recombinant human DNA fragmentation factor 45 kDa subunit (DFF45/ICAD), an inhibitor of the caspase activated DNase DFF40/CAD, substituted for healthy cytosol in inhibiting DNA fragmentation. An antiserum against human DFF45 detected 44 and 34 kDa proteins (major and minor, respectively) in the cytosols but not in the nuclear or membrane fractions of various cultured human cells. Cytosols depleted of DFF45/ICAD by immunoadsorption had little or no inhibitor of nuclear DNA fragmentation activity and no caspase-activated DNA fragmentation activity. We conclude that immunoreactive DFF45/ICAD is the principal inhibitor of apoptotic DNase activity in the cytosol of healthy cells. PMID- 9875237 TI - Regulation of multifunctional Ca2+/calmodulin-dependent protein kinases by Ca2+/calmodulin-dependent protein kinase phosphatase. AB - We have recently reported a novel protein phosphatase which dephosphorylates and thereby deactivates Ca2+/calmodulin-dependent protein kinase II (CaMKII) activated through autophosphorylation (Ishida, A., Kameshita, I., and Fujisawa, H. (1998) J. Biol. Chem. 273, 1904-1910). In the present study, we show that this protein phosphatase also catalyzed dephosphorylation of Ca2+/calmodulin-dependent protein kinases I (CaMKI) and IV (CaMKIV) which had been phosphorylated and activated by Ca2+/calmodulin-dependent protein kinase kinase alpha, resulting in reversible deactivation of the enzymes. The fairly high degree of the substrate specificity of this protein phosphatase suggests that the physiological significance of the phosphatase may be the regulation of the three multifunctional Ca2+/calmodulin-dependent protein kinases, CaMKI, CaMKII, and CaMKIV, which are the key enzymes in a Ca(2+)-signaling system in the cell. PMID- 9875238 TI - A novel ecto-phosphatase activity of Herpetomonas muscarum muscarum inhibited by platelet-activating factor. AB - In the present work ecto-phosphatase activity in Herpetomonas muscarum muscarum has been characterized using live parasites. This enzyme hydrolyzed p nitrophenylphosphate at a rate of 4.27 nmol Pi/mg of protein.min. A pH curve was generated, in which these intact flagellates showed the highest phosphatase activity at pH 6.5. Classical inhibitors for acid phosphatase, such as sodium orthovanadate, sodium tartrate, and ammonium molybdate, were used in the experiments and showed different patterns of inhibition. Lithium fluoride, aluminum chloride, and fluoroaluminate complexes were also tested. Although lithium fluoride and fluoroaluminate complexes were capable of inhibiting the phosphatase activity, aluminum chloride stimulated this enzyme. Cytochemical analysis showed the localization of this enzyme on the parasite surface. This ecto-phosphatase activity was also significantly diminished when the parasites were treated with 10(-6) M platelet-activating factor (PAF), a potent phospholipid mediator that promoted cellular differentiation in this parasite. PMID- 9875239 TI - Molecular cloning and functional expression of a fifth-type alpha 2,3 sialyltransferase (mST3Gal V: GM3 synthase). AB - The cDNA encoding a new type of alpha 2,3-sialyltransferase (mST3Gal V) was cloned from mouse brain cDNA library by PCR-based cloning approach using a pair of degenerate primers deduced from the nucleotide sequence information of mouse ST3Gal III and IV. The predicted amino acid sequence of mST3Gal V showed 27.3% and 26.4% identity to mST3Gal III and IV, respectively. The recombinant soluble mST3Gal V fused with protein-A, which expressed in the culture media of COS-7 cells, showed activity toward lactosylceramide (LacCer), and synthesized GM3. The apparent Km value for LacCer was 9.3 microM. mST3Gal V did not exhibit any activity toward other substrates we tested in this study, including glycolipids, glycoproteins and disaccharides. The mST3Gal V cDNA transfected F28-7 cells, which express large amount of lactosylceramide and very small amount of GM3 at native stage, expressed a large amount of GM3. The ST3Gal V gene was strongly expressed in mouse brain and liver, which contained a large amount of ganglioside. The gene expression seemed to be coincident with ganglioside expression in mouse. Thus, we conclude that mST3Gal V is the fifth-type alpha 2,3 sialyltransferase carrying GM3 synthetic activity. PMID- 9875240 TI - Identification of a binding site on Hsc70 for the immunosuppressant 15 deoxyspergualin. AB - Hsc70, the constitutive form of the heat shock protein 70 family of proteins, is involved in a number of biological activities which include protein folding and molecular chaperoning. Previously, we had shown that the immunosuppressant 15 deoxyspergualin (DSG) specifically interacted with Hsc70, as well as the Hsp90 family of proteins. Although the exact binding site on Hsc70 for protein substrates is unknown, a recent study shows that the extreme C-terminal four amino acids 647EEVD650 play a role in regulating AT-Pase activity, substrate binding, and interaction with HDJ-1. These four amino acids are also found at the C-terminus of Hsp90 and may be involved in similar functions. In this study, we show that DSG binds specifically to this EEVD regulatory domain. Binding of DSG to Hsc70 did not affect its ability to bind peptides. These results suggest that in addition to the ATP binding domain, there are two additional substrate binding domains on Hsc70. DSG should provide a tool for understanding the role of the EEVD motif in biological processes. PMID- 9875241 TI - Silica-induced pulmonary inflammation in rats: activation of NF-kappa B and its suppression by dexamethasone. AB - The goal of this study was to examine the relationship of the transcriptional regulatory factor nuclear factor-kappaB (NF-kappa B) to the early inflammatory events involved with silica exposure. Male F-344 rats received an intratracheal (i.t.) instillation of silica (100 mg/kg in a volume of 1 ml/kg) of saline. At 1, 3, 6, and 18 h postinstillation, and the rats were sacrificed and underwent bronchoalveolar lavage (BAL) for functional analysis of inflammation. Beginning at 1 h postinstillation, the silica-instilled (Si) rats displayed significant increases in neutrophils in BAL fluid compared to the saline controls. BAL cells from the Si group displayed a significant increase in luminol-dependent chemiluminescence (LDCL) compared to the controls. NF-kappa B activation was measurable at 3 h postinstillation, and this activation continued throughout the 18-h time course. Treatment with dexamethasone (5 mg/kg) at -3 h prior to silica instillation, at the time of instillation (0 h), and +1.5 h postinstillation resulted in both a reduction in NF-kappa B expression (by 70%) at 3 h postinstillation and corresponding reductions in LDCL, BAL cell count, and BAL neutrophils. These results show that activation of NF-kappa B is associated with silica-induced pulmonary inflammation, and the inhibition of its activation correlates temporally with suppression of inflammation. PMID- 9875242 TI - Potentiation of apoptosis by mitochondria in a cell-free system. AB - Using a cell-free system, we show that rat liver mitochondria, but not mitochondrial extracts, potentiated apoptosis triggered by cytosols derived from apoptotic cells. Apoptosis potentiated by mitochondria appeared to be inhibited by caspase 3 but not by caspase 1 inhibitors. A cytosolic caspase-3-like activity was increased by the addition of mitochondria to apoptotic cytosols; the latter activation was inhibited by the addition of bcl-2. Chelation of calcium by EGTA significantly and specifically inhibited the apoptosis potentiated by mitochondria as well as the increase of caspase-3-like activity. The incubation of mitochondria with apoptotic cytosols led to the release of cytochrome c, this latter phenomenon being inhibited by EGTA. Calcium or cytochrome c and dATP, however, did not reproduce the mitochondrial potentiation in the absence of the organelle. Thus, mitochondria can initiate and potentiate apoptosis through similar but not identical mechanisms. PMID- 9875243 TI - The androgen receptor CAG repeat polymorphism and its relationship to prostate cancer. PMID- 9875244 TI - Review of the 1998 Keystone Symposium on Transcriptional Mechanisms. Taos, NM, USA, February 21-26, 1998. PMID- 9875245 TI - An insight into the life of p53: a protein coping with many functions! Review of the 9th p53 Workshop, Crete, May 9-13, 1998. PMID- 9875246 TI - 1st MRC Human Genetics Symposium: maintenance of genomic stability. AB - This meeting served to juxtapose the fundamental studies on the distinct pathways which maintain genomic stability with work that addresses the phenotypic consequences of loss of genomic stability in humans. This created an exciting environment where we were prompted to think about the links between fundamental and applied research. It was also a forum where new ideas could be formed that will hopefully fuel interesting research in human disease. As we place the genome projects into perspective, the ideas arising from meetings such as the 1st MRC Human Genetics Symposium might be expected to guide studies that will reveal the molecular defects which underlie some of the more impenetrable phenotypes of human diseases. PMID- 9875247 TI - Transcription factors and cell differentiation. 20th Annual Symposium, Burnham Institute, La Jolla, CA, USA, 22 May 1998. PMID- 9875248 TI - Genetic medicine comes out of the cold in Sweden. Report on Nobel Workshop. 'Gene Targeted Drugs: Function and Delivery' held at Stockholm, 3 April 1998, and Friiberghs Herrgard, 4-5 April, 1998. PMID- 9875249 TI - The impact of hissy fits in primary care. AB - BACKGROUND: Hissy fits are experienced by physicians and patients alike, yet their full impact has never been studied before. METHODS: Specially trained researchers observed hissy fits at a clinic over 12 months. They interviewed perpetrators, victims and witnesses and recorded their comments because they had to. RESULTS: Hissy fits were common at the clinic and sometimes escalated to riots. Seasonal variations were endured. INTERPRETATION: Nobody likes this behaviour. Efforts should be made to counsel hissy fitters in channeling their angst in other, more positive ways. PMID- 9875250 TI - The good, the bad and the ugly. Association between car colour and bicycle passing space. AB - OBJECTIVE: Because anecdotal evidence indicates that the behaviour of cars (and their drivers) with respect to bicycles is highly variable, this study was undertaken to determine whether car colour correlates with the space allowed by the driver for passing a bicycle. DESIGN: Randomized recollection. SETTING: The streets of Vancouver and Burnaby, BC. PARTICIPANTS: The author, her bike, lots of cars and a few transit buses. METHODS: For a 10-day period in the summer of 1998, the investigator attempted, while cycling, to remember car colours and associated behaviours until she reached her various destinations. Data were eventually recorded in a tattered spiral-bound notebook saved from university days. OUTCOME MEASURES: Numbers of cars in 2 categories: "good" (those that gave extra space to cyclists) and "bad" (those that didn't). RESULTS: Read the article to find out. CONCLUSION: Although there was a slightly greater chance that a passing car would give a cyclist extra space, riders should be especially cautious when they catch sight of white and maroon vehicles. PMID- 9875251 TI - Cost-effectiveness of beer versus red wine for the prevention of symptomatic coronary artery disease. AB - BACKGROUND: Until now, red wine has been the cardioprotective beverage of choice, but the convenience and lack of adverse effects make beer a more cost-effective, safe and widely available choice. A paucity of evidence in support of this led to our study. METHODS: During a 6-month period, 11,780 consecutive patrons at 18 randomly selected urban and community liquor distributors were recruited. Participants were randomly assigned to receive either beer or wine daily for 5 years. Of the 10,078 people who met the inclusion criteria, 5033 were assigned to receive red wine and 5039 to receive beer every day. RESULTS: Effectiveness and adverse effects were equal in the 2 groups. However, costs for beer were substantially lower. INTERPRETATION: In view of its effectiveness, low cost, acceptable adverse-event profile and the fact that it is available in convenient single-dose dispensers, beer should replace red wine as the cardioprotective beverage of choice. PMID- 9875252 TI - The genetic basis of administosis. PMID- 9875253 TI - Evidence for a new classification of anal-retentiveness: torpid posteriosus and retro-pudendal hesitancy. AB - The introduction of ICD-10 has led to considerable debate over the proper diagnosis and treatment of retro-pudendal hesitancy (RPH). Many feel that the diagnosis should apply to all those with anal-retentiveness (AR). However, the authors discovered new clinical and laboratory evidence showing that AR is but one type of RPH. As such, they propose a reclassification of AR as type I RPH, a condition involving the superior pudendal-geniculate nucleus (SPGN). In contrast, type II RPH--another lesion of the SPGN--produces the characteristic torpid posteriosus (TP). PMID- 9875254 TI - Interviews with the dead. Using meta-life qualitative analysis to validate Hippocrates' theory of humours. AB - BACKGROUND: Hippocrates devised his theory of the 4 humours (blood, phlegm, black bile and yellow bile) 24 centuries ago. Since then, medicine has evolved into a complex body of confusing and sometimes contradictory facts. The authors, seeing a need to determine the validity of his theory, hired a psychic. METHODS: The psychic interviewed 4 eminent ancient physicians, including Hippocrates. A randomized double-blind cross-over design was used for this meta-life qualitative analysis. RESULTS: All of the interviewees agreed that the theory of humours is an accurate model to explain disease and personality. INTERPRETATION: Hiring a psychic to conduct after-death interviews with key informants is a useful way to validate scientific theories. PMID- 9875255 TI - Hissy fits revividus. PMID- 9875256 TI - The judgement of urines. PMID- 9875257 TI - McGee, Samuel: consultation report from the marge of Lake Lebarge. PMID- 9875258 TI - How to read clinical journals: IX. PMID- 9875259 TI - The holiday literature: a critical review. PMID- 9875260 TI - Katz on the Net. PMID- 9875262 TI - VASE-containing N-CAM isoforms are increased in the hippocampus in bipolar disorder but not schizophrenia. AB - The neural cell adhesion molecule (N-CAM) is a cell recognition molecule that is involved in cellular migration, synaptic plasticity, and CNS development. In schizophrenia, a 105- to 115-kDa N-CAM protein is increased in CSF and in the hippocampus and prefrontal cortex. The variable alternatively spliced exon (VASE) of N-CAM is developmentally regulated and can be spliced into any of the major 120-, 140-, and 180-kDa N-CAM isoforms. We determined that the variable alternative spliced exon of N-CAM (VASE) also is increased in bipolar disorder by quantitative Western immunoblot. VASE immunoreactive proteins (triplet bands around 140 kDa and a single band around 145 kDa) were identified in soluble and membrane brain extracts and quantified in the hippocampus. Soluble VASE 140 kDa was increased in the hippocampus of patients with bipolar disorder as compared to controls, patients with schizophrenia, and suicide cases. Membrane-extracted VASE 140 and 145 kDa were unchanged in the same groups. Multiple 145-kDa VASE immunoreactive proteins that also reacted to an N-CAM antibody were separated by isoelectric focusing and electrophoresis followed by western immunoblotting; however, the VASE 140-kDa proteins were only weakly N-CAM immunoreactive. By immunohistochemistry, VASE colocalized with GFAP-positive astrocytes in the hippocampus. VASE immunostaining was also observed in the cytoplasm of CA4 pyramidal neurons that were positive for phosphorylated high molecular weight neurofilament and synaptophysin terminals. Thus no differences in VASE were found in patients with schizophrenia, but there was a marked increase of VASE immunoreactive proteins in bipolar disorder. It is possible that abnormal regulation of N-CAM proteins results in differing patterns of abnormal expression in neuropsychiatric disorders. PMID- 9875263 TI - Regeneration of brainstem-spinal axons after lesion and immunological disruption of myelin in adult rat. AB - We previously observed that the transient developmental suppression of myelination or disruption of mature myelin, by local intraspinal infusion of serum complement proteins along with a complement-fixing, myelin-specific antibody (e.g., anti-Galactocerebroside), facilitated avian brainstem-spinal axonal regeneration after spinal transection. We now report the effects of similar immunological protocols on axonal regeneration in the injured adult rat spinal cord. After a lateral hemisection injury of the T10 spinal cord, infusion of the above reagents, over 14 days at T11, facilitated the regeneration of some brainstem-spinal axons. The hemisection lesion enabled comparisons between the retrograde labeling within an injured brainstem-spinal nucleus and the uninjured contralateral homologue. The brainstem-spinal nucleus examined in detail was the red nucleus (RN), chosen for its relatively compact descending pathway within the dorsolateral cord. Comparing the number of labeled neurons within each RN, of an experimentally myelin suppressed animal, indicated that approximately 32% of injured rubrospinal projections had regenerated into the caudal lumbar cord. In contrast, control-treated animals (e.g., PBS vehicle alone, GalC antibody alone, or serum complement alone) showed little or no axonal regeneration. We also examined the ultrastructural appearance of the treated cords. We noted demyelination over 1-2 segments surrounding the infusion site (T11) and a further two segments of myelin disruption (delamination) on either side of the demyelinated zone. The demyelination is an active process (< 3 days) with microglia and/or macrophages engulfing myelin. Thus, the facilitation of axonal regeneration through the transient suppression of CNS myelin may be fundamental to all higher vertebrates. PMID- 9875264 TI - Modifications of oligodendroglial cells in spongiform encephalopathies. AB - Although gray matter lesions involving neurones and astrocytes are prominent in human transmissible spongiform encephalopathies (TSE), white matter lesions have also been occasionally observed. Secondary (Wallerian) degeneration and direct myelin damage have been invoked, but the physiopathology of white matter involvement is still debated. We performed an immunohistochemistry study with anti-PrP antibodies of autopsy material of four patients with Creutzfeldt-Jakob disease (CJD), together with transmission electron microscopy (TEM) studies of conventionally processed biopsy specimens of the same patients. Light microscopy immunolabeling was observed as arrays adjacent to myelinic fibers and as a clumps adjacent to oligodendroglial nuclei; both cerebrum and cerebellum were involved. At the ultrastructural level, two types of intracellular inclusions were seen in the white matter. They were associated with dense lysosomes in oligodendroglial perikarya and in their processes. The inclusions were made of finely fibrillar, paracrystalline, amorphous, or densely osmophilic material. Thus, our findings may suggest that white matter involvement in spongiform encephalopathy is due to direct modifications of oligodendroglial cell associated with abnormal metabolism of PrP. PMID- 9875265 TI - Striatal transplantation in a transgenic mouse model of Huntington's disease. AB - Striatal grafts have been proposed as a potential strategy for striatal repair in Huntington's disease, but it is unknown whether the diseased brain will compromise graft survival. A transgenic mouse line has recently been described in which hemizygotes with an expanded CAG repeat in exon 1 of the HD gene exhibit a progressive neurological phenotype similar to the motor symptoms of Huntington's disease. We have therefore evaluated the effects of the transgenic brain environment on the survival, differentiation, and function of intrastriatal striatal grafts and undertaken a preliminary analysis of the effects of the grafts on the development of neurological deficits in the host mice. Hemizygote transgenic and wild-type littermate female mice received striatal grafts at 10 weeks of age and were allowed to survive 6 weeks. Normal healthy grafts were seen to survive and differentiate within the striatum of transgenic mice in a manner comparable to that seen in control mice. The transgenic mice exhibited a progressive decline in body weight from 9 weeks of age and a progressive hypoactivity in an open field test of general locomotor behavior. Although striatal grafts exerted a statistically significant influence on several indices of this impairment, all behavioral effects were small and did not exert any clinically relevant effect on the profound neurological deficiency of the transgenic mice. PMID- 9875266 TI - Changes of NGF presence in nonneuronal cells in response to experimental allergic encephalomyelitis in Lewis rats. AB - We recently reported that the cerebrospinal fluid (CSF) of patients affected by multiple sclerosis (MS) and the brain tissues of rats with experimental allergic encephalomyelitis (EAE) contain elevated levels of nerve growth factor (NGF). In the present study, we demonstrate that astrocytes and oligodendrocytes particularly localized in the white matter, including corpus callosum, overexpress NGFmRNA and produce NGF protein in the CNS of EAE affected rats. These findings indicate that the increased NGF found in the brain of EAE rats and most probably also in the CSF of patients affected by MS is produced by activated glial cells. It is hypothesized that the enhanced production of NGF by glial cells is necessary to compensate for the effect of axonal and/or neuronal cell body injury occurring in EAE. The possible functional significance of these findings in demyelinating diseases is discussed. PMID- 9875267 TI - Depletion of intracellular zinc induces protein synthesis-dependent neuronal apoptosis in mouse cortical culture. AB - The central nervous system (CNS) contains a large amount of zinc; a substantial fraction of it is located inside synaptic vesicles of glutamatergic terminals in chelatable forms and released in a calcium-dependent manner with intense neuronal activity. Recently, it has been shown that excessive zinc influx can kill neurons in rats subjected to transient forebrain ischemia. On the other hand, severe depletion of zinc has been also reported to induced cell death in certain nonneuronal cells. Since decreases in tissue zinc have been associated with Alzheimer's disease (AD) and senile macular degeneration, we examined whether depletion of intracellular zinc with a zinc chelator can directly induce neuronal death in mouse cortical cultures. Exposure of cortical cultures to a cell permeant zinc-chelator, N,N,N',N'-tetrakis (2-pyridylmethyl) ethylenediamine (TPEN, 0.5-3.0 microM) induced gradually developing neuronal degeneration accompanied by various features of apoptosis: cell body shrinkage, nuclear condensation and fragmentation, and internucleosomal DNA breakage. At higher concentrations, TPEN induced additional glial cell death. TPEN-induced cell death was completely blocked by coaddition of zinc. Addition of a protein synthesis inhibitor cycloheximide as well as a caspase inhibitor carbobenzoxy-valyl-alanyl aspartyl-fluoromethyl ketone (zVAD-fmk) markedly attenuated TPEN-induced neuronal death. On the other hand, brain-derived neurotrophic factor (BDNF), insulin-like growth factor-1 (IGF-1), phorbol 12-myristate 13-acetate (PMA), high K+, or an antioxidant, trolox, did not show any protective effect. The present results demonstrated that depletion of intracellular zinc induces protein synthesis dependent neuronal apoptosis in cortical culture. Combined with the findings that extracellular zinc may promote extracellular beta-amyloid (A beta) aggregation and that total tissue zinc is reduced in AD, present results suggest a possibility that redistribution of zinc from intracellular to extracellular space may synergistically contribute to neuronal apoptosis in AD. PMID- 9875268 TI - VEGF mRNA and its receptor flt-1 are expressed in reactive astrocytes following neural grafting and tumor cell implantation in the adult CNS. AB - Significant angiogenesis occurs only after injury in the adult mammalian brain; capillaries proliferate and astrocytes are activated by presently unresolved cellular mechanisms. Because of the intimate relationship between astrocytes and brain capillaries we examined the expression of the specific endothelial mitogen vascular endothelial growth factor (VEGF) in reactive astrocytes following CNS trauma models: neural grafting, stab wounds, and glioma implantation. In situ hybridization was combined with GFAP immunohistochemistry to delineate VEGF mRNA expression in reactive astrocytes. In addition, VEGF and its receptor flt-1 protein expression were detected immunohistochemically. In all three models we found unexpectedly that only reactive astrocytes, not endothelium, expressed the VEGF receptor flt-1, VEGF mRNA, and VEGF protein in a spatiotemporal manner, suggesting that activated astroglia may have a direct role in the induction of angiogenesis or permeability in mature brain. In addition, secreted VEGF may play a part in astroglial signalling by the induction of its own receptor in reactive astroglia following injury. These findings may have significant implications with regard to growth and reparative mechanisms of the adult cerebrovasculature. PMID- 9875269 TI - NK1, NMDA, 5HT1a, and 5HT2 receptor binding sites in the rat lumbar spinal cord: modulation following sciatic nerve crush. AB - Quantitative receptor binding autoradiography was used to study the NK1, NMDA, 5HT1a, and 5HT2 receptor binding densities in the adult rat lumbar spinal cord from 3 days to 20 weeks following a unilateral crush lesion of the sciatic nerve. NK1 binding density increased unilaterally in the superficial dorsal horn on the side of the sciatic crush to reach levels 60% above controls by 4 weeks following the lesion and returned to control values by 12 weeks. NMDA binding density increased bilaterally and equally in both the dorsal and ventral horns to reach 300% of control values at 2 weeks following the crush and returned to near control values by 20 weeks following the lesion. Serotonergic receptor binding did not change. The changes in NK1 receptor binding density on postsynaptic dorsal horn cells are consistent with a response to the decrease and recovery in the synthesis and transport of tachykinins by the dorsal root ganglion cells following peripheral nerve injury. the bilateral changes in NMDA receptor binding are more likely mediated by polysynaptic pathways in the spinal cord that respond to the changes in metabolic events of the dorsal root ganglion cells evoked by axotomy and regeneration. PMID- 9875270 TI - Motor and somatosensory deficits following uni- and bilateral lesions of the cortex induced by aspiration or thermocoagulation in the adult rat. AB - We have previously shown that lesions of the sensorimotor cortex induced by either thermocoagulation or aspiration produce different effects on axonal plasticity. We have now investigated whether these methods of lesion also influence the behavioral outcome. The behavioral effects of unilateral and bilateral lesions of the sensorimotor cortex induced by either aspiration or by thermocoagulation of pial blood vessels were examined in adult Sprague-Dawley rats. Rats were tested to determine limb use asymmetry by analyzing (1) coordinated forelimb placement and (2) paw use preference when rearing. Their responsiveness to somatosensory stimulation was tested by analyzing (1) the latency to remove sticky tape on the ventral surface of the paw, and (2) vibrissae-stimulated forelimb placing. Behavioral tests were performed prior to surgery and on day 4, 8, 12, 16, and 20 after surgery. Both unilateral lesions resulted in an over-reliance on the nonimpaired forelimb as early as 4 days after the surgery; functional recovery occurred after 16 days. Animals with bilateral lesions did not use either forelimb for support in postural support behaviors. However, this effect was more apparent in the animals with a thermocoagulatory lesion and, in contrast to the animals with an aspiration lesion, these animals did not show functional recovery. Animals with a unilateral aspiration, but not a thermocoagulatory lesion, showed a slowed response to tactile stimulation applied to the contralateral forelimb. After bilateral lesions, animals showed a slowed response to tactile stimulation applied to either forelimb at early time points after the lesion and recovery of function at later time points. These data indicate that, for the most part, lesions of the sensorimotor cortex by aspiration or thermocoagulation produce very similar effects on the behaviors examined in this study. However, unexpectedly, thermocoagulatory lesions induced a more severe (unilateral lesion) or prolonged (bilateral lesion) deficit in forelimb use than aspiration lesions. Conversely, the effect on tactile stimulation is more prominent after unilateral aspiration than thermocoagulatory lesions. PMID- 9875271 TI - Inflammation and Alzheimer's disease: relationships between pathogenic mechanisms and clinical expression. AB - During the past 15 years a variety of inflammatory proteins has been identified in the brains of patients with Alzheimer's disease (AD) postmortem. There is now considerable evidence that in AD the deposition of amyloid-beta (A beta) protein precedes a cascade of events that ultimately leads to a local "brain inflammatory response." Here we reviewed the evidence (i) that inflammatory mechanisms can be a part of the relevant etiological factors for AD in patients with head trauma, ischemia, and Down's syndrome; (ii) that in cerebral A beta disorders the clinical symptoms are determined to a great extent by the site of inflammation; and (iii) that a brain inflammatory response can explain some poorly understood characteristics of the clinical picture, among others the susceptibility of AD patients to delirium. The present data indicate that inflammatory processes in the brain contribute to the etiology, the pathogenesis, and the clinical expression of AD. PMID- 9875272 TI - Long acellular nerve transplants for allogeneic grafting and the effects of basic fibroblast growth factor on the growth of regenerating axons in dogs: a preliminary report. AB - Sciatic nerves were excised from 3 beagle dogs about 5 h after their sacrifice, treated three times by freezing and thawing, and stored in physiological saline for 3 months at -20 degrees C until used. Nerve segments 5 cm in length prepared from these stored nerves were transplanted to the common peroneal nerve in the right hindlimb of beagle dogs. Sixteen beagle dogs in total were used, in four treatment groups of two pairs each studied at 1 and 3 months. Five-hundred microliters basic fibroblast growth factor (bFGF) of two different concentrations (10 micrograms/300 microliters and 100 micrograms/300 microliters) which were impregnated in 0.5 ml gelatin hydrogels was applied around the sutured allografts. Autografting was also done in 4 beagle dogs, with no bFGF application. One month after the grafting, no regenerating nerves extended beyond the middle of the transplant in any of the allografts, except in the autografts in which a number of regenerated (myelinated) axons were present. Three months after the grafting, an abundance of myelinated axons was found at the middle of the graft: the numbers of axons per 10(4) micron 2 were 22.6 in the autografts and 10.6, 10.4 and 19.2 in the allografts treated with no bFGF, low-dose bFGF, and high-dose bFGF, respectively. Regenerating axons extended into the host nerve: the numbers of myelinated axons at the level 1.5 cm distal to the distal suture were 35.7, 0.9, 3.8, and 12.1 per 10(4) micron 2 in the above respective order. Although it was inferior in quality to the autograft, peripheral nerve regeneration was extensive in the distal nerve using freeze-thawed and bFGF treated allografts at 3 months. Electromyography showed that the peroneus longus muscle responded to the electrical stimuli given at the site proximal to the transplant in all four groups. These data indicate that a 5-cm acellular nerve segment containing Schwann cell basal laminae can be used successfully as an allograft without any immunosuppressants and that exogenously applied bFGF can improve nerve regeneration by enhancing the growth of regenerating axons. PMID- 9875273 TI - The development of the synaptic organization of the serotonergic system differs in brain areas with different functions. AB - The serotonergic innervation of the developing superior colliculus and ventrolateral nucleus of the thalamus of the rat were studied with light and electron microscope immunocytochemistry. We compared the pattern of innervation and synaptic organization of the serotonin (5-HT) system in the superficial and deep layers of the superior colliculus. We also compared the developmental pattern of synaptic incidence of 5-HT varicosities in the superior colliculus with that in the ventrolateral nucleus. Serotonin fibers were present in the superior colliculus at birth, concentrated mainly in the deep layers, whereas the superficial layers were only sparsely innervated. By the end of the first postnatal week the overall density of 5-HT fibers increased, but was still higher in the deep than in the superficial layers. The distribution pattern, density, and morphology of serotonergic axons acquired mature features by the end of the third postnatal week. In the adult, these axons were thin, varicose, forming a complex network which was denser in the lower part of the superficial layers and the upper part of the deep layers. Electron microscopical analysis revealed that the vast majority of 5-HT varicosities established symmetrical synapses with dendritic shafts in all layers of the superior colliculus throughout development. In the superficial layers, known to be involved in visual functions, the proportion of varicosities forming synapses increased gradually from birth to reach a peak at the end of the first postnatal week, then declined markedly in the subsequent 2 weeks before rising again at later stages. In contrast, in the deep layers and in the ventrolateral nucleus of the thalamus, areas involved in motor functions, the proportion of 5-HT varicosities engaged in synaptic contacts showed a continuous increase from birth until adulthood. Considering these results together with data from our previous studies, we speculate that the regional heterogeneity in the synaptic organization of the serotonergic system may reflect a differential role of 5-HT in the development of brain areas with different functions. PMID- 9875274 TI - Progressive changes in cortical water and electrolyte content at three stages of rat infantile hydrocephalus and the effect of shunt treatment. AB - Infantile hydrocephalus causes injury to the developing brain and despite surgical treatment, neurological deficits persist. The H-Tx rat develops inherited hydrocephalus in late gestation. Rapid postnatal ventricular enlargement, results in severe hydrocephalus by 21 days after birth. This is accompanied by changes in cortical morphology and metabolite content that indicate possible changes in intracellular composition. This study has tested the hypothesis that tissue water and electrolyte content is altered in hydrocephalus. The objective was to gain further insight into the mechanisms leading to neuronal damage. Water and electrolyte content (Na+, Cl-, and K+) were measured in the cerebral cortex of control and hydrocephalic rats at 4, 11, and 21 days after birth, and at 21 days in rats that received alleviating shunt surgery at 4 or 11 days. At all ages, hydrocephalic tissue was significantly increased over control for cortical water, Na+, and Cl- content. Additionally, at the intermediate (11 day) and advanced (21-day) stages there were significant decreases in K+ content, consistent with previous observations of decreases in organic osmolytes and energy metabolites. This suggests that by 11 days there are intracellular changes, probably through impaired membrane homeostatic mechanisms. In shunt treated rats, the extracellular constituents were almost normal, although a small increase over control values persisted. The decrease in intracellular K+ was not corrected in either group of shunt-treated rats. It is concluded that early hydrocephalus is characterized by extracellular edema that largely reverses with shunt treatment. Subsequently, as the hydrocephalus progresses, there is a breakdown of cell homeostasis and an irreversible loss of intracellular constituents. PMID- 9875275 TI - Regional difference in epidermal thinning after skin denervation. AB - Denervation of skin has a profound influence on epidermis; epidermal thinning was a consistent finding in rats. However, it is not clear whether the degree of epidermal thinning was similar in the region receiving the same innervation. In mice, how early epidermal nerves were degenerated after nerve injury remained unknown. To address these issues, we transected the sciatic nerve in mice and compared the changes of epidermal thickness in different areas of the hind foot skin. Epidermal nerves degenerated within 48 h after nerve transection, similar to what was observed in rats. Seven days after nerve transection, there was differential thinning of epidermis. The interpad area, in the center of the sciatic nerve-innervated region, exhibited the most profound degree of epidermal thinning (34.6 +/- 3.1 vs 47.8 +/- 2.4 microns, P < 0.01). The heel area, in the periphery of the sciatic nerve-innervated zone, did not show significant thinning of epidermis after denervation (37.3 +/- 4.8 vs 41.5 +/- 5.1 microns, P > 0.05). The degree of epidermal thinning after denervation in the pad area was the intermediate one: with 98.8 +/- 4.8 vs 120.1 +/- 7.3 microns, P < 0.02, in the rete pegs, and 51.1 +/- 4.1 vs 62.1 +/- 6.0 microns, P < 0.02, in the dermal papilla. The differential thinning was obvious when the thickness of the denervated epidermis was normalized to that of the control epidermis with the ratios of 0.73 +/- 0.03 in the interpad area, 0.83 +/- 0.04 in the rete peg, 0.85 +/- 0.05 in the dermal papilla, and 0.92 +/- 0.05 in the heel. Epidermal thinning was reversed by reinnervation of the epidermis after sciatic nerve crush (41.5 +/ 1.5 vs 45.0 +/- 2.0 microns in the interpad area, P > 0.05). These findings suggest that sensory nerves exhibit trophic influences on the epidermis presumably through the effects of diffusible factors. PMID- 9875276 TI - [125I]EGF binding in basal ganglia of patients with Parkinson's disease and progressive supranuclear palsy and in MPTP-treated monkeys. AB - Since EGF is known to protect and stimulate the activity of dopaminergic neurons, an autoradiographic study of [125I]EGF binding sites was performed in the striatum and pallidal complex in parkinsonian syndromes. The analysis was performed on postmortem brain tissues of three control subjects, three patients with Parkinson's disease, and three patients with progressive supranuclear palsy, another parkinsonian syndrome in which dopaminergic neurons also degenerate. Since all six patients had been treated with L-Dopa, we also analyzed the effects of this drug in an animal model of Parkinson's disease. Quantitative analysis of [125I]EGF binding was performed on the brains of three control monkeys, nine monkeys rendered parkinsonian by MPTP intoxication, three of which were treated with L-Dopa. An increased density of [125I]EGF binding was observed at anterior levels in the dorsal striatum, but not in the pallidum, of patients with Parkinson's disease and progressive supranuclear palsy. [125I]EGF binding was unchanged in parkinsonian monkeys whether or not they had been treated with L Dopa. The data suggest an increased expression of EGFRs in the striatum in chronic parkinsonian syndromes but not in acute models of the disease. PMID- 9875277 TI - Cell death, axonal damage, and cell birth in the immature rat brain following induction of hydrocephalus. AB - We hypothesized that hydrocephalus can cause death of brain cells and that generation of new brain cells might compensate for the cell loss. Hydrocephalus was induced in 3-week-old rats by injection of kaolin into the cisterna magna. The brains were studied 1 to 4 weeks later by histochemical, immunochemical, and ultrastructural methods. The ventricles enlarged progressively. Some axons in the corpus callosum were injured as early as 1 week, but axonal damage was not prevalent until 4 weeks when ventriculomegaly became severe. Dying cells detected by DNA end labeling and often identified as oligodendrocytes by electron microscopy were evident in white matter. Late-stage hydrocephalus was associated with a significant increase in the quantity of dying cells. Hydrocephalus was associated with increased Ki67 labeling and bromodeoxyuridine incorporation in the subependymal zone. Reactive changes were identified among astrocytes, oligodendroglia, and microglia. We conclude that hydrocephalus causes, in addition to axonal injury, gradual cell death in the cerebrum, particularly the white matter. The brain response includes production of new glial cells, but whether the new cells play any beneficial role remains unknown. PMID- 9875278 TI - Brain-derived neurotrophic factor stimulates hindlimb stepping and sprouting of cholinergic fibers after spinal cord injury. AB - Neurotrophic factors have been proposed as a therapeutic treatment for traumatic brain and spinal cord injury. The present study determined whether exogenous administration of one such factor, brain-derived neurotrophic factor (BDNF), could effect behavioral recovery and/or histopathological changes after spinal cord injury. Adult rats received a mild or moderate contusion injury or complete transection of the mid-thoracic spinal cord. Immediately thereafter, they were infused intrathecally with vehicle or BDNF for 28 days. Behavioral recovery was evaluated for 6 weeks after injury, at which time the rats were sacrificed and the spinal cord tissue was examined histologically. The infusion of BDNF resulted in acute stimulation of hindlimb activity. These effects included activation of alternating airstepping in injured rats when the hindlimbs were unloaded as well as slight improvements in the rate of recovery in open field locomotion score. BDNF infusion was also associated with enhanced growth of cholinergic fibers at the injury epicenter, but did not affect white matter sparing or density of serotonergic axons at or below the injury site. Based on immunohistochemical detection of BDNF protein distribution, these described effects are likely to be mediated by the activation of cells and axons within the central injury region and the along the peripheral rim of the spinal cord. Together, these findings demonstrate that the exogenous infusion of BDNF after spinal trauma can influence postinjury outcome through mechanisms that include acute stimulation of hindlimb activity and neuritogenesis at the injury site. PMID- 9875279 TI - Quenching revisited: low level direct current inhibits amygdala-kindled seizures. AB - We have reported that low frequency stimulation (1 Hz for 15 min), applied after kindling stimulation of the amygdala, inhibited the development and expression of amygdala-kindled seizures, an effect we termed quenching. Subsequently, we discovered that this effect could only be achieved when certain stimulators were used that also emitted a low-level direct current (DC). The studies reported here indicate that DC, applied once daily for 15 min at intensities of 5-15 microA, produced an intensity-related attenuation of kindling development and an increase in the afterdischarge threshold. This effect persisted in some animals for at least 1 month after discontinuation of the DC. In fully kindled animals, a robust increase in seizure threshold and persistent seizure inhibition were also observed using 10 microA of DC administered for 14 days. These results clarify and extend our original findings of a quenching effect; however, the mechanisms by which low level DC induces quenching require further elucidation. PMID- 9875281 TI - An integrated analysis of the progression of cell responses induced by permanent focal middle cerebral artery occlusion in the rat. AB - Defining the chronology and severity of cell damage in an evolving lesion after ischemia is important for understanding the underlying mechanisms in the development of therapeutic intervention. In the present study, we used a combination of histological and immunocytochemical methods to evaluate cell responses from 30 min to 48 h after permanent occlusion of the middle cerebral artery (MCAO) in the rat. Specific immunocytochemical markers clearly revealed acute early responses in neurons (neurofilament protein 200), astrocytes (glial fibrillary acidic protein), and microglia/macrophages (OX-42 and ED-1) such as enlarged, convoluted neuronal processes, and disintegration of glia. Progressive topographic changes in the developing lesion, pinpointed by immunolabeling, indicated the severity and extension of the cell damage. Proliferation and hypertrophy of astrocytes and microglia around the infarct, and contralaterally, occurred 24-48 h after MCAO and coincided with mass necrosis and infiltration of neutrophils and macrophages into the core. These observations corroborate the suggestion that the inflammatory process is involved in the progression of the infarct. PMID- 9875280 TI - Overexpression of HA-Bax but not Bcl-2 or Bcl-XL attenuates 6-hydroxydopamine induced neuronal apoptosis. AB - Bax, a member of the Bcl-2 gene family, is known to promote apoptosis in many cases but to block cell death under certain conditions. To investigate the potential role of Bax in 6-hydroxydopamine (6-OHDA)-induced cell death, we first established and characterized a dopaminergic neuronal cell line (MN9D) stably overexpressing hemagglutinin epitope-tagged Bax (MN9D/HA-Bax) as well as control clones (MN9D/Neo). Treatment of MN9D/Neo cells with 6-OHDA induced typical apoptotic cell death accompanied by shrinkage of the cell, nuclear condensation, and DNA fragmentation as demonstrated by light microscopy and agarose gel analysis. Overexpression of HA-Bax in MN9D cells was shown to attenuate 6-OHDA induced cell death as determined by the MTT reduction assay and agarose gel analysis for DNA fragmentation. Western blot analysis revealed that cleavage of poly(ADP-ribose)polymerase induced by 6-OHDA was attenuated in MN9D/HA-Bax cells. In contrast, overexpression of a well-known cell death-inhibiting protein such as Bcl-2 or Bcl-XL did not attenuate 6-OHDA-induced cell death. Interestingly, cell death induced by hydrogen peroxide (0.25-2.0 mM) was significantly accelerated, whereas the rate of cell death induced by menadione (10-50 microM) was not affected in MN9D/HA-Bax cells. Thus, our present data suggest that the functionally diverse roles of Bax may be determined by the type of stress applied to the cell. PMID- 9875282 TI - Alterations in GABAA receptor alpha 1 and alpha 4 subunit mRNA levels in thalamic relay nuclei following absence-like seizures in rats. AB - Modification of GABAA receptor mRNA levels by seizure activity can regulate general neuronal excitability. The possibility of absence seizure-induced alteration in GABAA receptor alpha 1, alpha 4, beta 2, and gamma 2 subunit gene expression in thalamic relay nuclei was studied in a rat model of absence seizures induced by gamma-hydroxybutyric acid (GHB). We observed a marked increase in alpha 1 mRNA and a corresponding decrease in alpha 4 mRNA in thalamic relay nuclei 2-4 h after the onset of GHB-induced absence seizures (when the seizures were terminating). These changes were selective to these alpha isoforms as neither beta 2 nor gamma 2 mRNA changed following seizures and occurred only in thalamic relay nuclei but not in hippocampus, a structure from which absence seizures do not evolve. The alterations in alpha 1 and alpha 4 mRNA persisted until about 12 h, and by 24 h after the seizure-onset the mRNA levels normalized. Blocking GHB-seizures produced no change in the levels of alpha 1 and alpha 4 mRNA in thalamic relay nuclei, suggesting that seizures themselves were responsible for mRNA alterations. In order to determine if absence seizure induced changes in alpha 1 and alpha 4 mRNA had any physiological significance, GHB was readministered in rats 6 and 24 h after the onset of seizures. The total duration of GHB-seizures was found to be significantly decreased when GHB was readministered at 6 h but not 24 h after the seizure-onset. These results suggest that absence seizures regulate GABAA receptor alpha 1 and alpha 4 gene expression in thalamic relay nuclei as a compensatory mechanism by which absence seizures are terminated. PMID- 9875283 TI - Changes in brain-derived neurotrophic factor immunoreactivity in rat dorsal root ganglia, spinal cord, and gracile nuclei following cut or crush injuries. AB - In the present study, we evaluated changes in brain-derived neurotrophic factor (BDNF) immunoreactivity in the rat lumbar (L) 5 dorsal root ganglion (DRG) and areas where afferents from the DRG terminate, the L5 spinal cord and gracile nuclei, following unilateral sciatic nerve transection or crush. From 3 days to 4 weeks following cut or crush injury, the percentage of medium and large BDNF immunoreactive neurons in the ipsilateral DRG increased significantly compared with those on the contralateral side. Following cut injury, there was no significant change in the percentage of small BDNF-immunoreactive neurons in the ipsilateral DRG; however, the intensity of immunoreactivity of these cells decreased. Following crush injury, however, both the percentage and intensity of small BDNF-immunoreactive neurons in the ipsilateral DRG significantly increased. Following cut injury, the expression of BDNF-immunoreactive axonal fibers decreased markedly in the ipsilateral superficial laminae of the L5 spinal cord and increased significantly in the ipsilateral deeper laminae of the spinal cord and gracile nuclei. Crush injury induced a marked increase in the expression of BDNF-immunoreactive axonal fibers in the superficial laminae of the spinal cord and gracile nuclei. These differences in BDNF response in the DRG and spinal cord after cut or crush injuries may reflect differences in trophic support to the injured DRG neurons and altered neuronal activity in the spinal cord and gracile nuclei following different types of peripheral nerve injury. PMID- 9875284 TI - Differential blood-brain barrier breakdown and leucocyte recruitment following excitotoxic lesions in juvenile and adult rats. AB - Acute neuronal degeneration can be induced by intracerebral injections of the glutamate receptor agonists kainic acid (KA) and NMDA (N-methyl-D-aspartate). It is accompanied by an inflammatory response that has not yet been fully investigated. We have previously demonstrated that the juvenile rat brain is more susceptible to an inflammatory challenge when compared to adult rat brain. This study set out to investigate whether this also applied to the inflammatory response associated with acute neuronal degeneration. NMDA and kainic acid were injected into the rat striatum and lesion size, leucocyte recruitment, and blood brain barrier (BBB) breakdown were assessed after 4, 8, 12, 24, 72, and 168 h. Both NMDA and KA induced lesions of similar volume at either age and apoptotic and necrotic nuclei could be detected. NMDA induced cellular loss by 4 h, whereas KA-injected rats did not show signs of neuronal loss until 8-12 h. The inflammatory response was characterized by an infiltration of neutrophils followed by macrophages. Juvenile rats showed a greater susceptibility to leucocyte recruitment compared to adult rats. BBB breakdown in response to NMDA injection occurred in the absence of cellular recruitment at 4 h in juveniles and was significantly greater in juvenile compared to adult rats at 8 h. BBB breakdown was minimal in KA-injected animals while at 7 days an influx of serum IgG coincided with a loss of astrocytic GFAP staining within the lesion. PMID- 9875286 TI - Mutators in Escherichia coli. PMID- 9875285 TI - Distinct roles for sodium, chloride, and calcium in excitotoxic dendritic injury and recovery. AB - The postsynaptic neuronal dendrite is selectively vulnerable to hypoxic-ischemic brain injury and glutamate receptor overactivation. We explored the glutamate receptor pharmacology and ionic basis of rapid, reversible alterations in dendritic shape which occur in cultured neurons exposed to glutamate. Dendrite morphology was assessed with the fluorescent membrane tracer, DiI, or immunofluorescence labeling of the somatodendritic protein, MAP2. Cortical cultures derived from 15-day-old mouse embryos underwent segmental dendritic beading when exposed to NMDA, AMPA, or kainate, but not to metabotropic glutamate receptor agonists. Varicosity formation in response to NMDA or kainate application was substantially attenuated in reduced sodium buffer (substituted with N-methyl-D-glucamine). Furthermore, veratridine-induced sodium entry mimicked excitotoxic alterations in dendrites and additionally caused varicosity formation in axons. Solutions deficient in chloride (substituted with Na methylsulfate) and antagonists of chloride-permeable GABA/glycine receptors reduced NMDA- or kainate-induced varicosity formation. An increase in dendrite volume was observed as varicosities formed, and varicosity formation was attenuated in sucrose-supplemented hypertonic media. Despite marked structural changes affecting virtually all neurons, dendrite shape returned to normal within 2 h of terminating glutamate receptor agonist application. Neurons exposed to kainate recovered more rapidly than those exposed to NMDA, and neurons exposed to NMDA in calcium-free buffer recovered more rapidly than cells treated with NMDA in normal buffer. While sodium, chloride, and water entry contribute to excitotoxic dendritic injury acutely, calcium entry through NMDA receptors results in lasting structural changes in damaged dendrites. PMID- 9875287 TI - Eukaryotic mismatch repair: an update. AB - The discovery that mutations in mismatch repair genes segregate with hereditary nonpolyposis colon cancer has awakened a great deal of interest in the study of the process of postreplicative mismatch repair. The characterisation of the principal players involved in this important metabolic pathway has been greatly facilitated by the amino acid sequence conservation among functional homologues of bacteria, yeast and mammals. The phenotypes of mismatch repair deficient mutants are also similar in many ways. In humans, mismatch repair malfunction demonstrates itself in the form of a mutator phenotype of the affected cells, an instability of microsatellite sequences and increased levels of somatic recombination. Moreover, mismatch repair deficient cells display also varying levels of tolerance to DNA damaging agents and are thought to be involved in the cell killing mediated by these agents. This article discusses some recent developments in this fast-moving field. PMID- 9875288 TI - Expression of the human XPB/ERCC-3 excision repair gene-homolog in the sponge Geodia cydonium after exposure to ultraviolet radiation. AB - The marine demosponge Geodia cydonium encodes a gene, termed GCXPB, which displays 62% identity to the human XPB/ERCC-3 gene that specifically corrects the repair defect in xeroderma pigmentosum and in Cockayne's syndrome. The cDNA was isolated and characterized the deduced aa sequence, XPB_GEOCY, with the calculated size of 91,541 Da comprises the characteristic domains found in the related helicases. Phylogenetic tree analysis revealed that the sponge sequence is grouped to the metazoan related XPB/ERCC-3 polypeptides. Northern Blot analyses have been performed with sponge samples collected at different depths, thus exposed to different intensities of UV sunlight in the field. The intensity of the 2.6 kb band, corresponding to the transcripts of the sponge GCXPB gene was highest in those biotopes, which are closer to the surface of the sea, lower were the expressions in animals from a cave or from depths of 22 to 35 m. Controlled laboratory studies revealed that after irradiation of specimens with 300 or 1000 J/m2 UVB light a dose-dependent increase of the steady-state level of GCXPB occurs, values up to 29-fold with respect to the controls which were kept in the dark have been determined. In parallel, the DNA integrity in the sponge samples was measured using the sensitive Fast Micromethod assay. The data revealed that the degree of strand DNA breaks paralleled the increase of expression of the GCXPB gene. From these data it is concluded that the XPB/ERCC-3-like gene in the sponge G. cydonium is UV light-inducible and hence might be used as biomarker for UV light exposure in the field. PMID- 9875289 TI - Efficient PCNA complex formation is dependent upon both transcription coupled repair and genome overall repair. AB - The protein proliferating cell nuclear antigen (PCNA) is an auxiliary factor for DNA polymerase delta and is involved in the resynthesis step of nucleotide excision repair (NER). After UV irradiation of quiescent cells, PCNA forms an insoluble complex with nuclear substructures. We have investigated associations between NER and its subcomponent pathway, transcription coupled repair (TCR) on PCNA complex formation using genetically related hamster cell lines with different repair characteristics. In DNA repair proficient cells, the PCNA complex was readily detectable within 30 min after UV irradiation by both immunofluorescence and western blot analyses. This complex formation after UV occurs efficiently in quiescent cells. In UV5 (human XP-D homolog) and UV 24 (human XP-B homolog) cells, which are totally deficient in NER, the PCNA complex was not detectable at 30 min after UV. The PCNA complex formation is restored to normal levels in UV5 cells after transfection with the human XPD gene, encoding a subunit of the basal transcription factor, TFIIH. In UV61 (Human CS-B homolog) cells, that are defective only in transcription coupled repair (TCR) of cyclobutane pyrimidine dimers (CPDs), the rate of PCNA complex formation was 2 fold slower than in repair proficient cells. This defect was complemented by transfection of the CSB gene into the UV61 cells. We thus conclude that efficient PCNA complex formation after UV is dependent upon both the NER and TCR pathways in hamster cells. The association of several other DNA repair proteins including XPA, RPA, TFIIH and p53 with the insoluble PCNA complex in UV treated cells suggests a central role for PCNA in different steps of NER. PMID- 9875290 TI - The C-terminal domain of p53 catalyzes DNA-renaturation and strand exchange toward annealing between intact ssDNAs and toward eliminating damaged ssDNA from duplex formation through preferential recognition of damaged DNA by a duocarmycin. AB - The C-terminal domain of p53 may bind single-stranded (ss) DNA ends and catalyze renaturation of ss complementary DNA molecules, suggesting a possible direct role for p53 in DNA repair (Proc. Natl. Acad. Sci. USA, 92, 9455-9459, 1995). We found that DU-86, a duocarmycin derivative which alkylates DNA, bound ssDNA and enhanced the DNA binding activity of the p53 C-terminus. DU-86 weakened p53 mediated catalysis of complementary ssDNA renaturation. p53 C-terminus catalyzed DNA strand transfer toward annealing between intact ssDNAs and toward eliminating DU-86-damaged ssDNA from duplex formation. These results suggest that p53, via the C-terminal domain, may play a direct role in DNA repair by preferential recognization and elimination of damaged DNA. PMID- 9875291 TI - The nature and origin of DNA single-strand breaks determined with the comet assay. AB - After treatment with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), methyl methanesulfonate (MMS) and hydrogen peroxide, the level of alkali-labile sites and single-strand breaks (ssb) in DNA was investigated, using the comet assay. The ability of antioxidant pre-treatment to decrease DNA damage was assessed. Results showed the following. (a) All single-strand (ss) DNA breaks detected immediately after MNNG- and MMS-treatment in hamster V79 cells had the character of alkali-labile sites while true ssb of DNA were represented only as a minor statistically significant (p < 0.01) fraction at the highest MMS concentration. (b) Most ss DNA breaks detected immediately after H2O2-treatment had the character of true breaks in DNA and alkali-labile sites represented only a minor fraction. (c) Pre-treatment of hamster V79 and human CaCo2 cells with vitamin E significantly reduced the number of breaks induced by hydrogen peroxide, but has no effect on the level of breaks induced by MNNG or MMS. We suggest that MNNG and MMS do not induce significant oxidative damage of DNA. Most of breaks induced by hydrogen peroxide have the nature of oxidative lesions of DNA. (d) In contrast to the effect of vitamin E, stobadine (STB) decreased not only the breaks induced by hydrogen peroxide but also those induced by MNNG and MMS. The reduced level of DNA damage in STB pre-treated samples could be due to inactivation of these alkylating agents by STB. PMID- 9875292 TI - The sensitization of cells treated with O6-methylguanine to alkylation damage is affected by the number of O6-methylguanine-DNA methyltransferase molecules escaped from inactivation. AB - O6-Methylguanine (MeG) can bind to the active site of O6-methylguanine-DNA methyltransferase (MGMT) as a free base. The subsequent methyl transfer reaction inactivates the repair protein. Hence, MeG is used to deplete the active MGMT pools in Chinese hamster cell lines (CHO) transfected to express varying amounts of human MGMT. After treatment with the free base, a residual population of active protein molecules remains localized mostly in the cytoplasm. Depleted cells are then challenged with the alkylating drug mitozolomide. Genotoxicity of this agent varied among the cell lines, and the compound sensitivity seemed to be regulated by a steady state equilibrium of residual MGMT molecules between nucleus and cytoplasm. PMID- 9875293 TI - Defective Kin28, a subunit of yeast TFIIH, impairs transcription-coupled but not global genome nucleotide excision repair. AB - The essential Saccharomyces cerevisiae KIN28 gene encodes a subunit of general transcription factor TFIIH, a multiprotein complex required for RNA polymerase II transcription initiation and nucleotide excision repair (NER). Kin28 is implicated in the transition from transcription initiation to transcription elongation by phosphorylation of the carboxy-terminal domain (CTD) of the largest subunit of the RNA polymerase II complex. Here, we explore the possibility that Kin28 like the other subunits of TFIIH is involved in NER in vivo, using yeast cells carrying either a wildtype or a thermosensitive KIN28 allele. The removal of UV induced cyclobutane pyrimidine dimers (CPDs) was monitored at base resolution from both strands of the RNA polymerase II transcribed genes RPB2 and URA3. Cells carrying the thermosensitive KIN28 allele display a transcription coupled repair (TCR) defect at the non-permissive temperature, which was most pronounced directly downstream of transcription initiation, probably as an indirect result of a general decrease in the level of RNA polymerase II transcription. The fact that CPD removal in non-transcribed DNA is completely unaffected in these cells indicates that Kin28 is not essential for general NER in vivo, providing the first example of a TFIIH subunit that is required for TCR but not for NER in general. PMID- 9875294 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin induction of cytochrome P450-dependent arachidonic acid metabolism in mouse liver microsomes: evidence for species specific differences in responses. AB - Arachidonic acid is biotransformed to metabolites active in signal transduction by cytochrome P450 (CYP) as well as by cyclooxygenase and lipoxygenase enzymes. Inducers of CYP1 enzymes, including 2,3,7,8-tetrachlorodibenzo-p-dioxin and other Ah receptor ligands, markedly increase liver microsomal CYP-dependent arachidonic acid epoxygenation in chicks but depress epoxygenation in rat liver microsomes where they elicit about twofold increases in formation of other CYP products, omega-1 to omega-4-OH arachidonic acid. These studies examined the effect of TCDD on metabolism of [1-14C]-labeled arachidonic acid by mouse liver microsomes. Mouse liver microsomes metabolized arachidonic acid exclusively by a CYP dependent mechanism as evidenced by lack of metabolism in the absence of NADPH and by formation of specific CYP-dependent metabolites. The major constitutive products were epoxygenase products (EETs and EET-diols) and omega-OH arachidonic acid. Treatment with TCDD increased formation of omega-2- to omega-4-OH arachidonic acid products 23-fold, formation of omega-1-OH arachidonic acid about 5-fold, and formation of epoxygenase products and HETEs each about twofold. In contrast, TCDD treatment decreased formation of omega-OH arachidonic acid by over 70%. EET-diols comprised a greater fraction of total epoxygenase products in mouse liver microsomes than has been found for liver microsomes of other species. The high EET-diol formation was attributable to a non-TCDD-inducible, EET epoxide hydrolase activity in mouse liver microsomes. For comparison, the effect of TCDD on [1-14C]-labeled arachidonic acid was examined in homogenates of spleen, an immune system target of TCDD. While levels of total [1-14C]-arachidonic acid metabolism were comparable in both tissues, virtually all of the metabolism by spleen was CYP-independent, and it was unaffected by TCDD. Western blotting experiments showed that TCDD-induced mouse Cyp1a1 and 1a2 share immunologic epitopes with chick CYP1A4 and 1A5. However, in immunoinhibition studies, an antibody to CYP1A5, the chick arachidonate epoxygenase, was ineffective against TCDD-induced arachidonic acid metabolism in mouse liver microsomes, suggesting that there are differences in the catalytic sites or tertiary structures of CYP1A5 and the CYP-enzyme catalyzing the TCDD-induced arachidonic acid metabolism in mouse liver. This study shows that the effects of TCDD of the profile of CYP dependent arachidonic acid metabolities and the amounts produced in mouse liver microsomes differ from other species. The findings suggest that species differences in CYP1A catalytic activities including the metabolism of arachidonic acid may contribute to species differences in sensitivity to TCDD toxicity. PMID- 9875295 TI - Some alkyl hydroxy benzoate preservatives (parabens) are estrogenic. AB - The inadvertent estrogenicity of certain synthetic chemicals, and their subsequent effects on the endocrine system of humans and wildlife, is of concern. In this paper we report findings from in vitro and in vivo (uterotrophic) studies which confirm that a range of alkyl hydroxy benzoate preservatives (parabens) are weakly estrogenic. In a receptor-binding assay, butylparaben was able to compete with 3H-estradiol for binding to the rat estrogen receptor with an affinity approximately 5 orders of magnitude lower than that of diethylstilboestrol, and between 1 and 2 orders of magnitude less than nonylphenol. In an in vitro yeast based estrogen assay, the four most widely used parabens (namely methyl-, ethyl-, propyl-, and butylparaben) were all found to be weakly estrogenic with the most potent (butylparaben) being 10,000-fold less potent than 17 beta-estradiol. The estrogenic activity of parabens was inhibited by 4-hydroxy tamoxifen in vitro, illustrating the requirement of these chemicals to interact with the estrogen receptor in order to activate the yeast. When administered orally to immature rats, the parabens were inactive. However, subcutaneous administration of butylparaben produced a positive uterotrophic response in vivo, although it was approximately 100,000 times less potent than 17 beta-estradiol. Given their use in a wide range of commercially available topical preparations, it is suggested that the safety in use of these chemicals should be reassessed, with particular attention being paid to estimation of the actual levels of systemic exposure of humans exposed to these chemicals. The acquisition of such data is a prerequisite to the derivation of reliable estimates of the possible human risk of exposure to parabens. PMID- 9875297 TI - Alteration in lung particle translocation, macrophage function, and microfilament arrangement in monocrotaline-treated rats. AB - Individuals with preexisting cardiopulmonary disease are thought to be more susceptible to acute episodes of particulate pollution resulting in increased morbidity and mortality. Our study was designed to evaluate particle fate and macrophage function in an animal model of monocrotaline (MCT)-induced pulmonary hypertension. Two weeks following a single MCT injection, Sprague-Dawley rats were exposed sequentially to two different colored fluorescent microspheres 1.0 micron in diameter by aerosolization. Morphometric evaluation of lung sections was performed 0 and 24 h following the final particle exposure to determine the intrapulmonary location of inhaled microspheres. A decrease in the number of particles phagocytized by alveolar macrophages and an increase of free particles overlying the epithelium were found in MCT-treated animals compared with control. Pulmonary macrophages recovered by bronchoalveolar lavage were evaluated for chemotactic and phagocytic ability. Macrophage chemotaxis was significantly impaired following MCT treatment compared with controls, whereas phagocytic activity of macrophages lavaged from MCT and control treatment groups was similar. Macrophages were stained for filamentous (F) and globular (G) actin using Texas-Red-labeled phalloidin and Oregon-green-labeled DNase I, respectively. The area of microfilament staining for F and G actin increased, but the ratio of F/G actin was significantly decreased in animals with MCT treatment compared with control. While the responses observed with MCT treatment, such as pulmonary edema, polymorphonuclear leukocytes influx, and unique macrophage morphology may contribute to impaired macrophage function, the change in microfilament arrangement suggests that MCT may inhibit macrophage chemotaxis and impair particle clearance from the lungs. PMID- 9875296 TI - Biotransformation of perchloroethene: dose-dependent excretion of trichloroacetic acid, dichloroacetic acid, and N-acetyl-S-(trichlorovinyl)-L-cysteine in rats and humans after inhalation. AB - Chronic exposure of rodents to perchloroethene (PER) increased the incidence of liver tumors in male mice and resulted in a small but significant increase in the incidence of renal tumors in male rats. The tumorigenicity of PER is mediated by metabolic activation reactions. PER is metabolized by cytochrome P450 and by conjugation with glutathione. Cytochrome P450 oxidation of PER results in trichloroacetyl chloride which reacts with water to trichloroacetic acid (TCA) which is excreted. The formation of S-(trichlorovinyl)glutathione (TCVG) from PER results in nephrotoxic metabolites. TCVG is cleaved to S-(trichlorovinyl)-L cysteine (TCVC) and acetylated to N-acetyl-S-(trichlorovinyl)-L-cysteine (N-ac TCVC), which is excreted with urine. TCVC is also cleaved in the kidney by cysteine conjugate beta-lyase to dichlorothioketene which may react with water to dichloroacetic acid (DCA) or with cellular macromolecules. The object of this study was to comparatively quantify the dose-dependent excretion of PER metabolites in urine of humans and rats after inhalation exposure. Three female and three male human volunteers and three female and three male rats were exposed to 10, 20, and 40 ppm PER for 6 h, and three female and three male rats to 400 ppm. A dose-dependent increase in the excretion of TCA and N-ac-TCVC after exposure to PER was found both in humans and in rats. A total of 20.4 +/- 7.77 mumol of TCA and 0.21 +/- 0.05 mumol of N-ac-TCVC were excreted in urine of human over 78 h after the start of exposure to 40 ppm PER; only traces of DCA were present. After identical exposure conditions, rats excreted 1.64 +/- 0.42 mumol of TCA, 0.006 +/- 0.002 mumol of N-ac-TCVC and 0.18 +/- 0.04 mumol of DCA. Excretion of N-ac-TCVC in male rats exposed to 400 ppm PER (103.7 nmol) was significantly higher, compared to female rats (31.5 nmol) exposed under identical conditions. N-ac-TCVC was rapidly eliminated with urine both in humans (t1/2 = 14.1 h) and in rats (t1/2 = 7.5 h). When comparing the urinary excretion of N-ac TCVC, a potential marker for the formation of reactive intermediates in the kidney, humans received a significantly lower dose (3 nmol/kg at 40 ppm) compared to rats (23.0 nmol/kg) after identical exposure conditions. In addition, rats excreted large amounts of DCA which likely is a product of the beta-lyase dependent metabolism of TCVC in the kidney. The obtained data suggest that glutathione conjugate formation and beta-lyase-dependent bioactivation of TCVC in PER metabolism is significantly higher in rats than in humans. Thus, using rat tumorigenicity data for human risk assessment of PER exposure may overestimate human tumor risks. PMID- 9875298 TI - Ketoconazole-induced apoptosis through P53-dependent pathway in human colorectal and hepatocellular carcinoma cell lines. AB - In this study, we first demonstrated that the widely used oral antifungal drug, ketoconazole (KT), can induce apoptosis in various type of human cancer cells and in a primary culture of rat liver cells. We further investigated the molecular mechanisms of KT-induced apoptosis. It was found that KT induced nuclear accumulation of p53 protein in a dose- and time-dependent manner. The level of p53 protein was elevated approximately three times as much in treated cells 24 h after KT (5 microM) exposure as in cells receiving mock treatment. We found that cells containing wild-type p53 (COLO 205 and Hep G2) were more sensitive to KT exposure. The bax protein was induced and the bcl-2 protein was inhibited by KT in cells containing wild-type p53 (Hep G2, COLO 205) but not in cells without p53 (Hep 3B). The caspase-3 was activated 24 h after KT treatment. The Poly-(ADP ribose) polymerase (PARP) and the lamin A degradation was induced by KT, which promoted nuclear membrane disassembly and eventually caused apoptosis. Our results also indicated that none of the PKC gene family was involved in KT induced apoptosis. PMID- 9875299 TI - Acute CdMT injection is not a good model to study chronic Cd nephropathy: comparison of chronic CdCl2 and CdMT exposure with acute CdMT injection in rats. AB - Kidney is the main target organ of Cd toxicity in humans. Cd-induced nephrotoxicity is thought to be caused by the Cd-metallothionein complex (CdMT) that "leaks" out of the liver and is taken up by the kidney. A single injection of CdMT has therefore been used as a model to study Cd nephropathy for the last 20 years. However, our recent studies reveal discrepancies between renal Cd concentration and nephrotoxic potencies of CdCl2 and CdMT. This study was further designed to critically evaluate whether a single injection of CdMT is an appropriate model to study the mechanism of chronic CdCl2 nephropathy. Age matched rats were given multiple sc injections of either CdCl2 (0.8 and 1.2 mg Cd/kg) or CdMT (0.05 mg Cd/kg) daily, 6 days/week for 6 weeks, or a single injection of CdMT (0.2-0.6 mg Cd/kg i.p. for 24 h), and the nephrotoxicity was compared. Histologically, chronic CdCl2 or CdMT administration produced damage to the whole kidney, including tubular cell degeneration, apoptosis, and atrophy; interstitial inflammation; glomerular swelling; and sclerosis. In contrast, acute CdMT injection produced severe proximal tubule necrosis as the major feature of its toxicity. Biochemically, chronic exposure to Cd produced polyuria and calciuria, while proteinuria, glucosuria, and enzymuria were mild (2-5x). In contrast, acute CdMT nephrotoxicity was characterized by marked increases in urinary protein (13x), glucose (25x), N-acetyl-beta-d-glucosaminidase (28x), lactate dehydrogenase (100x), and gamma-glutamyltranspeptidase (160x). Serum levels of creatinine and blood urea nitrogen were unchanged following chronic Cd exposure but were markedly elevated (5x) after acute injection of CdMT. Chronic exposure to either CdCl2 or CdMT produced nephrotoxicity at renal Cd concentration of 85 to 110 micrograms/g kidney, while acute CdMT injection produced nephrotoxicity at only 5 to 7 micrograms/g kidney. In conclusion, the present study indicates that the features and mechanisms of renal injury from chronic Cd exposure are quite different from those produced by a single injection of CdMT. Therefore, it is proposed that acute CdMT injection is not an appropriate model for the study of chronic Cd-induced nephrotoxicity. PMID- 9875300 TI - Arachidonic acid metabolism in primary irritant dermatitis produced by patch testing of human skin with surfactants. AB - A clinical study was performed to determine the effects of patch testing human skin with four industrially used surfactants on erythema formation, transepidermal water loss, and the contents in suction blister fluids of primary proinflammatory mediators including arachidonic acid, eicosanoids, and IL-1 alpha, which were analyzed by quantitative gas chromatography/negative ion chemical ionization mass spectrometry and by an enzyme-immunoassay, respectively. Benzalkonium chloride (BKCI) and sodium lauryl sulfate (SLS) elicited erythema and caused increased transepidermal water loss, indicating a disturbance of the epidermal barrier. Triethanolamine (TEA) and Tween 80 did not evoke these gross symptoms of inflammation. Suction blister fluids collected after a 24-h application of BKCl, SLS, and Tween 80 contained significantly increased amounts of individual eicosanoids whereas TEA induced no response. The induced eicosanoid profile was characteristic for each compound, pointing to different cell types of skin to be involved in their production. The elevation of prostaglandin and LTB4 contents correlated with the induction of erythema and the impairment of the epidermal barrier as shown for BKCl and SLS and preceded the maximum of erythema formation. IL-1 alpha contents did not correlate with these gross symptoms of inflammation. The results of this in vivo study support those of a previous study using human keratinocytes in culture indicating the release of arachidonic acid and prostaglandins to be an early event involved in the interaction of keratinocytes with surfactants. Moreover, the in vivo data with human skin underscore the mechanistic relationship to the in vitro model and support the concept that arachidonic acid and eicosanoid release from keratinocytes can be used as a marker of primary skin irritation. PMID- 9875301 TI - The importance of fiber biopersistence and lung dose in determining the chronic inhalation effects of X607, RCF1, and chrysotile asbestos in rats. AB - The chronic inhalation effects in rats of X607 (a rapidly dissolving synthetic vitreous fiber) were compared with those previously reported for RCF1 (a refractory ceramic synthetic vitreous fiber) and chrysotile asbestos. Of primary concern was the importance of biopersistence as a mechanism of fiber toxicity. Fischer rats were exposed to fiber aerosol by nose-only inhalation for 6 h/day, 5 days/week for 2 years. X607 and RCF1 aerosols were similar in concentration (approximately 200 fibers/cc) and average dimensions (approximately 20 x 1 microns). Chrysotile aerosol was higher in concentration (10,600 fibers/cc) and an order of magnitude smaller in average dimensions. Lung fiber deposition after 6 h inhalation was greater for X607 than for RCF1. However, at later time points, fibers/lung (especially long fibers) were much lower for X607 than for RCF1, suggesting less biopersistence for X607. X607 was neither fibrogenic nor tumorigenic and induced only minimal lung cellularity that reversed after exposure was terminated. In contrast, RCF1 and chrysotile asbestos induced pulmonary fibrosis and thoracic neoplasms (chrysotile induced 32% more pulmonary neoplasms than RCF1). Lung deposition and fiber lengths did not explain the toxicologic differences between the three fibers. Fiber biodurability, including chemical and physical parameters, appears to be a major toxicologic determinant here. Chemical analysis of lung fibers revealed rapid leaching of X607 compared to RCF1. In in vitro dissolution tests, X607 underwent rapid dissolution (kdis = 990 ng/cm2/h) and transverse fragmentation, RCF1 dissolved slowly (kdis = 6 ng/cm2/h) and did not fragment, and chrysotile dissolution was negligible (< 0.1 ng/cm2/h). PMID- 9875302 TI - An integrated model for the differentiation of chemical-induced allergic and irritant skin reactions. AB - Contact and photocontact allergic as well as irritant and photoirritant skin reactions represent a major problem in clinical dermatology and during the development of new pharmaceuticals. Furthermore, there is a lack of in vitro and in vivo assays that provide a clear differentiation between allergic and irritant skin reactions. Here, we describe an integrated model to differentiate between chemical-induced allergic and irritant skin reactions by measuring objective and easy-to-determine parameters within both skin and skin-draining lymph nodes. Dose response studies with standard contact and photocontact allergens as well as irritants and photoirritants revealed that irritants predominantly induced skin inflammation, which in turn stimulated draining lymph node cell proliferation. In contrast, the induction phase of contact or photocontact allergy was characterized by marginal skin inflammation, but a marked activation and proliferation of skin-draining lymph node cells. Therefore, a differentiation index (DI) was defined describing the relation between skin-draining lymph node cell activation (lymph node cell count index) and skin inflammation (ear swelling). A DI > 1 indicates an allergic reaction pattern whereas DI < 1 demonstrates an irritant potential of a chemical. Experiments with the contact allergen oxazolone, the photocontact allergen TCSA + UVA, the irritant croton oil, and the photoirritant 8-methoxypsoralen + UVA confirmed the predictive value of DI. Furthermore, flow cytometric analysis of lymph node-derived T- and B-cell subpopulations revealed that contact sensitizer, but not irritant, induced the expression of CD69 on the surface of I-A+ cells. In conclusion, further studies with a broad range of irritants and allergens will be required to confirm general applicability. PMID- 9875303 TI - Mechanisms of Ca2+ antagonism in imipramine-induced toxicity of isolated adult rat cardiomyocytes. AB - Classic explanations of cyclic antidepressant toxicity often focus on Na+ channel blockade; however, cyclic antidepressant toxicity often causes decreased myocardial contractile function. The present experiments first examine inhibition of cytosolic Ca2+ signals by imipramine. Second, the experiments test if alkalinization prevents the inhibition of Ca2+ signals. Cardiomyocytes from adult rat hearts were loaded with fura-2 dye, and intracellular calcium, [Ca2+]i, was quantified using ratio fluorescence techniques. Changes in [Ca2+]I were induced by electrical pacing, depolarization with KCl (84 mM), or treatment with caffeine (10 mM). Imipramine (10-30 microM) inhibited [Ca2+]i transients in electrically paced cardiomyocytes. Imipramine (7.5-30 microM) also inhibited Ca2+ signals in KCl depolarized cells. These inhibitory effects were similar to those observed with nisoldipine (100-2000 nM), a selective L-channel blocker. The rise in [Ca2+]i that was triggered with caffeine (10 mM) was not significantly changed by imipramine (30 microM). Inhibition of KCl-induced Ca2+ signals by imipramine was prevented by alkalinization of the medium (tris(hydroxymethyl)aminomethane, pH 7.6), but not by elevation of extracellular sodium to 170 mM. Alkalinization was effective in the presence of HOE642, a selective Na+/H+ (NHE) subtype 1 inhibitor. These data show that imipramine causes Ca2+ antagonism in heart cells which is independent of sarcoplasmic reticulum Ca2+, and that alkaline treatment prevents this Ca2+ antagonism rather than stimulating an alternate source of Ca2+ via Na+/H+ and subsequent Na+/Ca2+ exchange. PMID- 9875304 TI - Role of CYP1A2 in the hepatotoxicity of acetaminophen: investigations using Cyp1a2 null mice. AB - Acetaminophen (APAP) is known to cause centrilobular hepatic necrosis under overdose conditions. This is thought to be mediated via the P450-generated reactive intermediate N-acetyl-p-benzoquinone imine (NAPQI). Initially, NAPQI is detoxified by conjugation with glutathione (GSH), but once GSH is depleted, NAPQI reacts more extensively with hepatic proteins leading to hepatocellular damage. The P450 isoforms thought to be responsible for APAP hepatotoxicity in humans are CYP2E1, CYP1A2, and CYP3A4, and thus, we have investigated the effect of murine Cyp1a2 on APAP hepatotoxicity using Cyp1a2 knockout mice (Liang et al., Proc. Natl. Acad. Sci. USA 93, 1671-1676, 1996). Doses of 250 mg/kg were markedly hepatotoxic in these mice, and surprisingly, deaths only occurred in the knock out and heterozygote mice over a 24-h period after dosing. Furthermore, there were no significant differences among survivors of any genotype in serum ALT concentrations, a well correlated indicator of APAP hepatotoxicity in mice. Finally, no differences were observed in the urinary metabolites excreted ove the 24-h period, including those derived from GSH conjugation of the major reactive metabolite NAPQI. Consistent with the effects on hepatotoxicity and metabolism, 2 h after hepatotoxic doses (500 mg/kg, i.p.) of APAP no significant differences were observed in total whole liver homogenate nonprotein thiol concentrations among the three genotypes even though hepatic thiols were decreased compared to control animals (> 90%). In addition, when the liver cytosol and microsome samples were examined by immunoblotting for the presence of APAP-protein adducts using a specific antiserum, there were no observable differences in either the intensity of staining or in the spectrum of adducts formed between APAP-dosed mice of any genotype. The cumulative data suggest that Cyp1a2 doses not play a significant role in APAP hepatotoxicity in these mice. PMID- 9875305 TI - Resistance to carbon tetrachloride-induced hepatotoxicity in mice which lack CYP2E1 expression. AB - CYP2E1 knockout mice (cyp2e1-/-) were used to investigate the involvement of CYP2E1 in the development of carbon tetrachloride (CCl4)-induced hepatotoxicity. Male cyp2e1-/- and wild-type (cyp2e1+/+) mice were given a single i.p. injection of 1 ml/kg (= 1.59 g/kg) CCl4 and 24 h later liver injury was assessed by elevations of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities and histopathology. No significant increases in serum ALT and AST activities were observed in cyp2e1-/- mice when compared to wild-type counterparts after CCl4 exposure. No detectable abnormality in liver histology was found in cyp2e1-/- mice after CCl4 exposure. In contrast, CCl4 treatment resulted in 442- and 125-fold increases in serum ALT and AST activities, respectively, in wild-type mice. Consistent with the results of serum ALT and AST activities, severe hepatic damage was noted in livers of wild-type mice, indicating the importance of CYP2E1 in mediating the hepatic damage following CCl4 exposure in these mice. In addition, a dramatic decrease in CYP2E1-catalyzed p-nitrophenol activity and complete loss of immunoreactive CYP2E1 were observed in wild-type mice after CCl4 treatment, suggesting that CYP2E1 was degraded during the process of CCl4-induced hepatotoxicity. These studies conclusively demonstrate that CYP2E1 is the major factor involved in the CCl4-induced hepatotoxicity in mice. PMID- 9875306 TI - Inhibition of protein phosphatase activity and changes in protein phosphorylation following acetaminophen exposure in cultured mouse hepatocytes. AB - Protein phosphorylation was determined in cultured mouse hepatocytes exposed to an hepatotoxic concentration of acetaminophen (APAP) for selected times up to 12 h. Cultures were radiolabled with 32P-orthophosphoric acid and the cell extracts were analyzed by 2D gel electrophoresis and autoradiography. APAP exposure selectively increased the phosphorylation state of proteins of molecular weight 22, 25, 28, and 59 kDa and decreased the phosphorylation of a 26-kDa protein. Evidence is presented that these changes (1) are dependent on cytochrome P-450 activation of APAP; (2) occur well before enzyme leakage in this in vitro model; (3) are not likely attributed to GSH depletion alone; (4) are in part mimicked by okadaic acid, calyculin A, and cantharidic acid, three structurally distinct inhibitors of protein phosphatases 1 and 2A; and (5) are paralleled by a decline in protein phosphatase activity. The physiological consequences of protein phosphatase inactivation could be significant in APAP overdose since these enzymes are involved in the dephosphorylation of regulatory proteins that control many cell functions. This study also provides the first evidence for disruption in signal transduction pathways as a response to or component of APAP-induced hepatic injury. PMID- 9875307 TI - U.S. Environmental Protection Agency's revised cancer guidelines for carcinogen risk assessment. PMID- 9875308 TI - Re: Organotin-induced apoptosis as observed in vitro is not relevant for induction of thymus atrophy at antiproliferative doses. (Article No. TO978265) PMID- 9875309 TI - The importance of anatomical realism for validation of physiological models of disposition in inhaled toxicants. PMID- 9875310 TI - Subgenomic RNA promoters dictate the mode of recognition by bromoviral RNA dependent RNA polymerases. AB - Both the brome mosaic virus (BMV) and cowpea chlorotic mottle virus (CCMV) RNA dependent RNA polymerases (RdRps) were found to recognize the BMV core subgenomic promoter in the same manner, requiring specific functional groups at positions 17, -14, -13, and -11 relative to the subgenomic initiation site (+1). For CCMV subgenomic RNA synthesis, both RdRps required the same nucleotides and four additional nucleotides at positions -20, -16, -15, and -10. The -20 nucleotide is partially responsible for the differential mode of recognition of the two promoters. These data provide evidence that the RNA can induce RdRps to alter the mode of promoter recognition. PMID- 9875311 TI - Importance of the intracytoplasmic domain of the simian immunodeficiency virus (SIV) envelope glycoprotein for pathogenesis. AB - SIVmac1A11 and SIVmac239 are nonpathogenic and pathogenic molecular clones in rhesus macaques, respectively. Although these viruses exhibit approximately 98% nucleotide and amino acid sequence homology, differences are found in the length of the translation frames for several genes. SIVmac239 has a premature stop codon in nef, whereas SIVmac1A11 has a premature stop codon in vpr and two premature stop codons in the intracytoplasmic domain of the env-transmembrane (TM) subunit. Recombinant viruses, constructed through reciprocal exchange of large DNA restriction enzyme fragments between SIVmac1A11 and SIVmac239, were evaluated in adult rhesus macaques. This in vivo analysis revealed that two or more regions of the SIVmac genome were essential for high virus load and disease progression (Marthas et al., 1993. J. Virol. 67, 6047-6055). An important gap in knowledge remaining from this study was whether the premature stop codons in env-TM of recombinant virus SIV1A11/239gag-env/1A11 (Full-length vpr and nef, two stop codons in env-TM) reverted to coding triplets in vivo. Here, we report that viral sequences in macaques, which succumbed to an AIDS-like disease after infection with SIV1A11/239gag-env/1A11, exhibited reversion of both env-TM stop codons. In addition, antibodies to the intracytoplasmic domain of env-TM were detected in macaques containing revertant virus and showing disease; this finding indicates that this domain of the env glycoprotein was expressed in vivo. Thus selection for viral variants with full-length env-TM demonstrated that the cytoplasmic domain of the SIVmac env glycoprotein plays a role in viral persistence and immunodeficiency in primates. PMID- 9875312 TI - Oncosis in MA104 cells is induced by rotavirus infection through an increase in intracellular Ca2+ concentration. AB - Rotavirus infection modifies the metabolism and ionic homeostasis of the host cell. First, there is an induction of viral synthesis with a parallel shutoff of cell protein production, followed by an increase of plasma membrane Ca2+ permeability, thereby inducing an increase of free cytoplasmic and sequestered Ca2+ concentrations. Cell death follows at a later stage. We studied the role of the increase in Ca2+ concentration in cell death. An elevation of extracellular Ca2+ concentration during infection induced an increase in [Ca2+]i and potentiated cell death. Buffering the increases in [Ca2+]i with BAPTA added at 6 h p.i. reduced the cytopathic effect without inhibiting viral protein synthesis and infectious particle production. Metoxyverapamil (D600), a Ca2+ channel inhibitor, added at 1 h p.i. reduced Ca2+ permeability, the increases in [Ca2+]i, and cell death produced by infection without modifying viral protein synthesis and infectious titer. Thapsigargin, the inhibitor of Ca(2+)-ATPase of endoplasmic reticulum, potentiated the increase of [Ca2+]i and accelerated the time course of cell death. Double staining with fluorescein diacetate and ethidium bromide or acridine orange and ethidium bromide showed that infected MA104 cells had lost plasma membrane integrity without DNA fragmentation or formation of apoptotic bodies. These results support the hypothesis that the increase in [Ca2+]i due to a product of viral protein synthesis triggers the chain of events that leads to cell death by oncosis. PMID- 9875313 TI - Human papillomavirus-induced carcinogenesis with p53 deficiency in mouse: novel lymphomagenesis in HPV16E6E7 transgenic mice mimicking p53 defect. AB - To investigate the transforming activity of human papillomavirus (HPV) E6 and E7 genes in vivo, we previously established transgenic mouse lines containing HPV16E6E7, in which male mice develop a Leydig cell tumors with a very high incidence. Because HPV-induced carcinogenesis is highly related to p53, we changed the dose of p53 gene in the transgenic lines by the mice crossing with p53-disrupted mice. The transgenic mice with homozygous wild-type p53 alleles developed only the testicular tumor, whereas novel T cell lymphomagenesis occurred in the heterozygous p53-disrupted E6E7 (p53+/-E6E7) transgenic mice. In this tumor and even in the normal spleen, the absence of p53 protein was observed, whereas the p53 mRNA was expressed with a normal size, suggesting the degradation of p53 protein in these tissues. These results suggest that HPV16E6 could stimulate p53 protein degradation in mouse cells and induced the lymphomagenesis in a manner indistinguishable from p53 deficiency. PMID- 9875314 TI - B cells malignantly transformed by human immunodeficiency virus are polyclonal. AB - The roles that human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV) play in the genesis of acquired immune deficiency syndrome (AIDS)-related lymphomas are not understood. A human B cell line (B-HIV), developed to study AIDS-related lymphomagenesis, contains EBV and HIV genomes and is malignantly transformed. This line was produced by exposing B cells from an EBV-seropositive donor to HIV. To investigate the number of independent transformation events that took place at the time of HIV infection, we sought to determine how many transformed lineages are present in this cell line. B-HIV was found to have multiple different sites of HIV integration (> or = 25) as determined by fluorescence in situ hybridization. As a control, we analyzed a clonal cell line of HIV-infected human T cells, 8E5, and found HIV sequences located exclusively at band q22 in chromosome 13. We conclude that B-HIV is polyclonal, and viral sequences are located at multiple variable chromosomal sites in different B-HIV cells. PMID- 9875315 TI - Mucosal immunisation with papillomavirus virus-like particles elicits systemic and mucosal immunity in mice. AB - It has been shown previously that recombinant virus-like particles (VLPs) of papillomavirus can induce VLP-specific humoral and cellular immune responses following parenteral administration. To test whether mucosal administration of bovine papillomavirus type 1 (BPV1) VLPs could produce mucosal as well as systemic immune responses to VLPs, 50 micrograms chimeric BPV1 VLPs containing an HPV16 E7 CTL epitope (BPVL1/E7 VLP) was administered intranasally to mice. After two immunisations, L1-specific serum IgG and IgA were observed. L1-specific IgG and IgA were also found in respiratory and vaginal secretions. Both serum and mucosal antibody inhibited papillomavirus VLP-induced agglutination of RBC, indicating that the antibody induced by mucosal immunisation may recognize conformational determinants associated with virus neutralisation. For comparison, VLPs were given intramuscularly, and systemic and mucosal immune responses were generally comparable following systemic or mucosal delivery. However, intranasal administration of VLP induced significantly higher local IgA response in lung, suggesting that mucosally delivered HPV VLP may be more effective for mediating local mucosal immune responses. Intranasal immunisation with HPV6b L1 VLP produced VLP-specific T proliferative responses in splenocytes, and immunisation with BPVL1 VLP containing an HPV16 E7 CTL epitope induced E7-specific CTL responses. We conclude that immunisation with papillomavirus VLPs via mucosal and intramuscular routes, without adjuvant, can elicit specific antibody at mucosal surfaces and also systemic VLP epitope specific T cell responses. These findings suggest that mucosally delivered VLPs may offer an alternative HPV VLP vaccine strategy for inducing protective humoral immunity to anogenital HPV infection, together with cell-mediated immune responses to eliminate any cells which become infected. PMID- 9875316 TI - Role of the transmembrane sequence of spleen focus-forming virus gp55 in erythroleukemogenesis. AB - The membrane glycoprotein encoded by the env gene of either the polycythemia- or anemia-inducing spleen focus-forming virus (SFFVp or SFFVa, respectively) is responsible for the induction of erythroleukemia in mice. It has been shown that the SFFVp glycoprotein, gp55, interacts with the erythropoietin receptor (EPO-R) and promotes EPO-independent proliferation of an EPO-R-expressing hematopoietic cell line, Ba/F3 (Li et al., Nature 343:762, 1990). We show here that when residues within the transmembrane (TM) sequence of an SFFVp gp55 are altered based on the sequences of the anemia-inducing gp55s by a methionine-to-isoleucine (M-I) substitution, a di-leucine deletion (dLL), or both, the resulting mutants display an attenuated phenotype that resembles an SFFVa: they induce milder erythroproliferative disease without polycythemia in vivo and are unable to promote EPO-independent cell proliferation in vitro. The dLL mutation directly interferes with EPO-R binding by decreasing the affinity of gp55 for the receptor. On the other hand, the M-I mutation hampers the full mitogenic activation of EPO-R while having no effect on receptor binding and asserts a dominant negative effect over the wild-type SFFVp gp55. Two other sequence changes within the TM sequence did not affect the biological activities of the SFFVp gp55. These results indicate that the TM sequence of the SFFV env glycoprotein plays a prominent role in SFFV-induced erythroleukemogenesis through its influence on the mitogenic activation of EPO-R. PMID- 9875317 TI - Phosphorylation of the M2 protein of influenza A virus is not essential for virus viability. AB - M2 is a minor component of the influenza A virus envelope. The cytoplasmic tail of the M2 protein is posttranslationally modified in the infected cell by palmitylation and phosphorylation. The primary site for phosphorylation of the M2 cytoplasmic tail is serine 64, which is highly conserved yet not required for the activity of the M2 ion channel. Using an exogenous incorporation assay, we have shown that incorporation of M2 into virus particles is type-specific and does not require phosphorylation of the cytoplasmic tail. In addition, phosphorylation of the cytoplasmic tail is not required for the directional transport of M2 in polarized MDCK cells. Using a reverse genetics and reassortment procedure, we generated a virus (Ra) specifically mutated in segment 7 such that the M2 cytoplasmic tail could no longer be phosphorylated. The virus was found to grow as well as wild-type virus in tissue culture and in eggs, was stable on passage in these systems, and possessed no second-site mutations in the engineered RNA segment. In vivo Ra replicated in Balb/c mice at least as well as the parent strain A/WSN/33. These studies indicate that phosphorylation of the M2 cytoplasmic tail is not required for in vitro or in vivo replication of influenza A virus. PMID- 9875318 TI - Characterization of the acidic domain of the IE1 regulatory protein from Orgyia pseudotsugata multicapsid nucleopolyhedrovirus. AB - This study presents a detailed analysis of the acidic N-terminal region of the Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV) transactivator IE1. The N-terminal region of IE1 is rich in acidic amino acids and has been hypothesized to be an acidic activation domain. Removal of the N-terminal 126 amino acids containing the acidic domain of IE1 resulted in complete loss of transactivation activity, indicating that this region is essential for transactivation. The OpMNPV acidic domain was replaced with the archetype acidic activation domain from VP16 and the acid-rich region of Autographa californica multicapsid NPV (AcMNPV) IE1. These chimeric constructs were fully capable of transactivation in transient assays. The chimeric OpMNPV IE1s containing the herpes simplex virus VP16 and AcMNPV IE1 acidic activation domains consistently transactivated a reporter gene to higher levels than the OpMNPV IE1 acidic activation domain. Transactivation by the chimeric constructs is enhanced synergistically when cotransfected with IE2 into Lymantria dispar and Spodoptera frugiperda cells. Both N- to C-terminal and C- to N-terminal deletions of the OpMNPV acidic activation domain were constructed to define functional domains within the OpMNPV IE1 acidic activation domain. At least two potential activation domains were identified. Within each of these domains, two core regions at amino acids 28-43 and amino acids 113-124 were identified that were similar to core regions of VP16 and GAL4, which contain predominately acidic and bulky hydrophobic amino acids. PMID- 9875319 TI - Nuclear factors that bind to the U3 region of two murine myeloid leukemia inducing retroviruses, Cas-Br-E and Graffi. AB - Cas-Br-E and Graffi are two myeloid leukemia-inducing murine viruses. Cas-Br-E induces, in NIH-Swiss mice, mostly non-T, non-B leukemia composed of very immature cells with no specific characteristics (Bergeron et al. (1993). Leukemia 7, 954-962). The Graffi murine leukemia virus causes exclusively myeloid leukemia, but the tumor cells are clearly of granulocytic nature (Ru et al. (1993). J. Virol. 67, 4722). We were interested to understand the role of the long terminal repeat (LTR) U3 region in the myeloid specificity of these two retroviruses. We used DNase I footprinting and gel mobility shift assays to identify a number of protein binding sites within Cas-Br-E and Graffi U3 regions. The pattern of protected regions is highly similar for the two viruses. Some factors identified in other murine leukemia viruses, like the core binding factor, also bind to Cas-Br-E and Graffi LTR; however, other binding sites seem specific for these two viruses. Only one difference between them was noted, at the 5' end of the U3 region. Transcriptional activity of both LTRs was also analyzed in various cell lines and compared with other murine leukemia viruses. The results show a slight myeloid specificity for the two LTRs, and indicate that the Graffi enhancer is quite strong in a broad range of cell types. PMID- 9875320 TI - The inhibitory effect of RANTES on the infection of primary macrophages by R5 human immunodeficiency virus type-1 depends on the macrophage activation state. AB - We investigated whether culture conditions could affect the RANTES antiviral effect on human immunodeficiency virus type 1 (HIV-1) infection of primary macrophages. Monocyte-derived macrophages (MDM) were obtained either as (1) the adherent cells of 5-day cultures of blood mononuclear cells (PBMC), followed by 2 days without nonadherent PBMC or added cytokines (MDM-5d), or (2) as the adherent cells recovered from 1-h incubation of PBMC, which were cultured for 7 days with macrophage colony-stimulating factor (M-CSF; MDM-MCSF). Infection of MDM-5d from different donors with HIV-1 R5 strains was reproducibly inhibited by RANTES (IC50 < or = 10 nM), but infection of MDM-MCSF was not inhibited by > or = 100 nM RANTES, even when added at initiation of cultures, although it was still inhibited by a CD4 antibody. RANTES had no antiviral effect when MDM-5d were treated with physiological concentrations of M-CSF or GM-CSF before infection. CCR5 and CXCR4 expression as well as that of other cell surface molecules, including adhesion molecules, was not affected by the cytokines. MDM-MCSF from delta 32CCR5 homozygous individuals did not render them permissive to HIV-1, suggesting that it is unlikely that the virus uses another coreceptor. RANTES binding to MDM was chondroitin sulfate, but not heparan sulfate, dependent, and RANTES bound more efficiently to MDM-5d than to MDM-MCSF. Chondroitin sulfate removal partially offset the RANTES antiviral effect for MDM-5d. Thus RANTES anti HIV-1 activity for primary macrophages depends on culture conditions and their consequent activation status, which may lead to differences in proteoglycan surface expression. These data may be relevant for the development of chemokine based therapy for HIV-1 infection. PMID- 9875321 TI - Localization and fine mapping of antigenic sites on the nucleocapsid protein N of porcine reproductive and respiratory syndrome virus with monoclonal antibodies. AB - The purpose of this study was to analyze the antigenic structure of the nucleocapsid protein N of the Lelystad virus isolate of porcine reproductive and respiratory syndrome virus (PRRSV) and to identify antigenic differences between this prototype European isolate and other North American isolates. To do this, we generated a panel of monoclonal antibodies (mAbs) directed against the N protein of Lelystad virus and tested them in competition assays with other N-specific mAbs described previously (Drew et al., 1995; Nelson et al., 1993; van Nieuwstadt et al., 1996). Four different competition groups of mAbs were identified. Pepscan analysis with solid-phase dodecapeptides was used to identify specific antigenic regions in the N protein that were bound by the mAbs. In this pepscan analysis, we found that the mAb of the first competition group reacted with linear peptides whose core sequences consisted of amino acids 2-12 (site A), the mAbs of the second group reacted with peptides whose core sequences consisted of amino acids 25-30 (site B), and the mAb of the third group reacted with peptides whose core sequences consisted of amino acids 40-46 (site C). However, the fourth group of mAbs binding to an antigenic region, provisionally designated as domain D, reacted very weakly or did not react at all with solid-phase dodecapeptides. To further characterize the structure of the epitopes in domain D, we produced chimeric constructs composed of the N protein sequences of Lelystad virus and another arterivirus lactate dehydrogenase-elevating virus, which was used because its N protein has similarity in amino acid sequence and hydropathicity profile but does not react with our mAbs. When the mAbs specific to domain D were tested for binding to the chimeric N proteins expressed by Semliki Forest virus, we found that the regions between amino acids 51-67 and amino acids 80-90 are involved in the formation or are part of the epitopes in domain D. Therefore, we conclude that the N protein contains four distinct antigenic regions. The epitopes mapped to sites A-C are linear, whereas the epitopes mapped to domain D are more conformation dependent or discontinuous. Sites A and C contain epitopes that are conserved in European but not in North American isolates; site B contains epitopes that are conserved in European and North American isolates; and site D contains epitopes that are either conserved or not conserved in European and North American isolates. The antigenic regions identified here might be important for the development of diagnostic test for PRRSV in particular tests that discriminate between different antigenic types of PRRSV. PMID- 9875322 TI - The herpes simplex virus 1 UL 17 gene is required for localization of capsids and major and minor capsid proteins to intranuclear sites where viral DNA is cleaved and packaged. AB - In nuclei of cells infected with herpes simplex virus (HSV), synthesized viral DNA accumulates as concatamers that are cleaved into genomic lengths and inserted into preformed capsids. Whereas newly replicated DNA and enzymes required for DNA synthesis accumulate in sites of infected cell nuclei termed replication compartments, the intranuclear site of DNA cleavage and packaging is currently controversial. DNA packaging requires the UL6, UL15, UL17, UL25, UL28, UL32, and UL33 genes in addition to the major capsid proteins. Using confocal immunofluorescence microscopy, it was observed that in > 95% of HEp-2 cells fixed at late times after infection with wild-type HSV-1, capsids, major capsid proteins ICP5 and ICP35, and the UL6-encoded minor capsid protein localized in DNA replication compartments. These data support the hypothesis that capsid assembly and DNA cleavage/packaging normally occur in HEp-2 cell replication compartments. In contrast, cells infected with a viral mutant lacking functional UL17 contained antigenically dense nuclear aggregates that stained with ICP35, ICP5, and capsid specific antibodies. Cells infected with the UL17 mutant virus also displayed UL6-specific fluorescence in a diffuse pattern at the nuclear periphery in regions not containing ICP35 and ICP5. Displacement of ICP35 from replication compartments was not observed in cells infected with cleavage/packaging mutants lacking UL28 and UL33. We conclude that the UL17 gene is required for correct targeting of capsids and major and minor capsid proteins to the DNA replication compartment of HEp-2 cells and deduce that this targeting reflects one functional role of UL17 in viral DNA cleavage and packaging. PMID- 9875323 TI - Intracellular trafficking and interactions of the HIV-1 Tat protein. AB - Fusions of the human immunodeficiency virus type 1 (HIV-1) transactivator protein Tat to the green fluorescent protein (GFP) were used to study the intracellular localization, trafficking, and interactions of Tat in human cells. Tagging Tat with GFP did not change its nuclear localization or ability to act as a transactivator. Tat-GFP expressed at low levels was found in the nucleus, whereas overexpression resulted in nucleolar accumulation. A Tat-GFP hybrid protein containing in addition the HIV-1 Rev nuclear export signal (NES) localized predominantly to the cytoplasm. This shuttle protein, Tat-GFP-NES, transactivated the HIV-1 long terminal repeat. Thus a Tat molecule being only transiently present in the nucleus is active and nucleolar accumulation of Tat is not prerequisite for function. A coexpression assay previously used to define protein interaction domains in the HIV-1 Rev protein [R. H. Stauber, E. Afonina, S. Gulnik, J. Erickson, and G. N. Pavlakis (1998a). Virology 251, 38-48.] indicated that Tat exists predominantly as a monomer and does not form stable multimers with B23 in living cells. Using a heterokaryon fusion assay, we found that Tat GFP was able to shuttle between the nucleus and the cytoplasm. Tat therefore has the potential to perform functions in the nucleus as well as in the cytoplasm. PMID- 9875324 TI - Role of cellular actin in the gene expression and morphogenesis of human respiratory syncytial virus. AB - Cytoskeletal protein actin and nonactin cellular proteins were essential for human respiratory syncytial virus (RSV) gene expression. In vitro, specific antibodies against actin inhibited RSV transcription, whereas antibodies against other cytoskeletal proteins had little or no effect. Affinity purified cellular actin or bacterially expressed recombinant actin activated RSV transcription. However, optimal transcription required additional cellular protein(s) that appeared to function as accessory factor(s) for actin. In the absence of actin, these proteins did not activate viral transcription. Purified viral nucleocapsids contained actin, but no cytokeratin, tubulin, or vimentin. Cytochalasin D or DNasel--agents that destabilize actin polymers--had little effect on RSV transcription. RSV infection itself seemed to alter the structure of the cellular actin filaments. Treatment of infected cells with cytochalasin D produced a more severe disruption of the filaments and drastically reduced the production of infectious virus particles but still had little effect on intracellular synthesis of viral macromolecules. Thus actin seems to serve a dual role in RSV life cycle: its monomeric form as well as polymeric form activate viral transcription, while only the microfilament form may take part in viral morphogenesis and/or budding. PMID- 9875325 TI - Characterisation of regulatory sequences at the Epstein-Barr virus BamHI W promoter. AB - Epstein-Barr virus, a human gammaherpesvirus, possesses a unique set of latent genes whose constitutive expression in B cells leads to cell growth transformation. The initiation of this growth transforming infection depends on a viral promoter in BamHI W (Wp) whose regulation is poorly understood. Using Wp reporter constructs in in vitro transfection assays, we found that Wp was 11- to 190-fold more active in B cell than in non-B cell lines and that three regions of the promoter (termed UAS1, UAS2, and UAS3) contributed to transcriptional activation. The upstream regions UAS3 (-1168 to -440) and UAS2 (-352 to -264) both functioned in a cell lineage-independent manner and were together responsible for the bulk of Wp activity in non-B cells; mutational analysis indicated the importance of a YY1 binding site in UAS2 in that context. By contrast, UAS1 (-140 to -87) was B cell specific and was the key determinant of the promoter's increased activity in B cell lines. Mutational analysis of UAS1 sequences combined with in vitro bandshift assays revealed the presence of three binding sites for cellular factors in this region. When mutations that abolished factor binding in bandshift assays were introduced into a Wp reporter construct, the loss of any one of the three UAS1 binding sites was sufficient to reduce promoter activity by 10- to 30-fold in B cells. From sequence analysis, two of these appear to be novel transcription factor binding sites, whereas the third was identified as a cyclic AMP response element (CRE). Our data indicate that this CRE interacts with CREB and ATF1 proteins present in B cell nuclear extracts and that this interaction is important for Wp activity. PMID- 9875326 TI - Comparison of the intranuclear distributions of herpes simplex virus proteins involved in various viral functions. AB - Herpesviral transcription, DNA synthesis, and capsid assembly occur within the infected cell nucleus. To further define the spatial relationship among these processes, we have examined the intranuclear distributions of viral DNA replication, gene regulatory, and capsid proteins using dual label immunofluorescence and confocal microscopy. We observed that several of the viral DNA replication proteins localize preferentially to punctate structures within replication compartments while the major transcriptional activator, ICP4, and the ICP27 regulatory protein show a more diffuse distribution within replication compartments. The viral proteins that show a punctate distribution in replication compartments redistribute from these compartments to prereplicative sites when viral DNA replication is inhibited, whereas viral proteins that show a diffuse distribution remain within replication compartments when viral DNA replication is inhibited. Thus the sites of viral DNA replication and late transcription appear to be distinct but codistribute within the boundaries of replication compartments. The major capsid protein, ICP5, also localizes initially to a diffuse distribution within replication compartments, but during the time of maximal progeny virus assembly, ICP5 becomes localized to punctate structures within replication compartments that are often near the punctate structures occupied by viral DNA replication proteins. Hence the processes of viral DNA replication, late transcription, and capsid assembly show a general overlapping distribution within replication compartments but appear to be located at distinct sites within these regions of the infected cell nucleus. PMID- 9875327 TI - Ebola virus inhibits induction of genes by double-stranded RNA in endothelial cells. AB - Fatal cases of filoviral infection are accompanied by a marked immunosuppression. Endothelial cells play a vital role in the host immune response through the expression of several immunomodulatory genes in addition to the expression of the antiviral genes, 2',5'-oligoadenylate synthetase [2'-5'(A)N], and the double stranded RNA (dsRNA)-activated protein kinase (PKR). dsRNA, an intermediate generated during viral replication and gene transcription of many viruses, leads to the induction of immunomodulatory genes in endothelial cells. In this report, we show that induction of the major histocompatibility complex class I family of genes, 2'-5'(A)N, interleukin-6 (IL-6), PKR, interferon (IFN)-regulatory factor 1, and intercellular adhesion molecule-1 (ICAM-1) by dsRNA in human umbilical vein endothelial cells is suppressed by infection with the filovirus Ebola-Zaire (EZ). In contrast, induction of IL-6 and ICAM-1 by IL-1 is intact in EZ-infected cells. Gel shift analysis demonstrates that dsRNA-induced protein binding to IFN responsive elements is strongly suppressed by EZ-IFN, whereas NF-kappa B activation by dsRNA remains intact. We previously reported that IFN signaling is suppressed by EZ infection, and these data strongly suggest that elements shared between IFN and dsRNA signaling are being inhibited by EZ. Inhibition of IFN and dsRNA responsiveness could play a role in the immunosuppression seen in EZ infections and would play a role in the pathogenesis of disease caused by EZ. PMID- 9875328 TI - Association between HTLV-1 Tax and I kappa B alpha is dependent on the I kappa B alpha phosphorylation state. AB - Biological, molecular, and epidemiological data have demonstrated that human T cell leukemia virus type 1 (HTLV-1) encoded Tax protein plays a central role in the initiation of T cell malignancy. The 40-kDa Tax oncoprotein serves as a potent transcriptional activator that induces viral gene expression driven by the HTLV-1 long terminal repeats and also stimulates multiple cellular genes involved in T cell activation, cell cycle regulation, and gene activation. Since Tax has been shown to interact directly and indirectly with the NF-kappa B/I kappa B regulatory proteins, we examined the significance of an in vivo association between Tax and the I kappa B alpha inhibitor. Using GST affinity chromatography, Tax was shown to interact with the I kappa B alpha ankyrin repeats which are essential for interaction with the NF-kappa B/Rel proteins. In vivo, using I kappa B alpha mutants and co-immunoprecipitation, a preferential interaction between HTLV-1 Tax and N-terminally hypophosphorylated I kappa B alpha was detected. Tax also enhanced binding of I kappa B alpha to the proteasome subunit HsN3, resulting in a Tax-enhanced, constitutive degradation of wild-type and mutated forms of I kappa B alpha in the absence of phosphorylation and ubiquitination. Binding of I kappa B alpha to proteasome subunit HC9 was also observed, but this interaction occurred independently of Tax. Taken together, these results suggest a role for Tax as a viral chaperone resulting in the enhanced constitutive turnover of I kappa B alpha. The association of Tax with hypophosphorylated I kappa B alpha may prevent I kappa B alpha from binding to NF kappa B and also target I kappa B alpha to the proteasome for degradation via a phosphorylation-independent pathway. PMID- 9875329 TI - Therapeutic periocular vaccination with a subunit vaccine induces higher levels of herpes simplex virus-specific tear secretory immunoglobulin A than systemic vaccination and provides protection against recurrent spontaneous ocular shedding of virus in latently infected rabbits. AB - Rabbits latently infected with herpes simplex virus type 1 (HSV-1) were vaccinated either periocularly or systemically with a subunit vaccine (gB2 + gD2) plus adjuvant or adjuvant alone. Tear films were collected daily to measure recurrent infectious HSV-1 shedding. After systemic vaccination, the latently infected rabbits were not protected against recurrent ocular viral shedding (HSV 1-positive tear film cultures/total cultures) compared with either the systemic or periocular adjuvant controls (systemic vaccination = 49 of 972, 5.0%; systemic control = 46 of 972, 4.7%; periocular control = 43 of 930, 4.6%; P > 0.8). In contrast, latently infected rabbits vaccinated periocularly with the same vaccine had significantly reduced recurrent shedding (20 of 1026, 2.0%) compared with controls (P < 0.001) or systemic vaccination (P = 0.0002). Thus, recurrent HSV-1 shedding was significantly reduced by therapeutic local periocular subunit vaccination but not by therapeutic systemic subunit vaccination. Neutralizing antibody titers in the serum of systemically and ocularly vaccinated rabbits was similar. In contrast, HSV-specific tear secretory immunoglobulin A was significantly higher in the ocularly vaccinated group (P < 0.01). These results strongly suggest that in the rabbit, and presumably in humans, the local ocular (mucosal) immune response is much more important than the systemic immune response for therapeutic protection against recurrent ocular HSV-1. Thus development of a therapeutic vaccine against recurrent ocular HSV-1 should be directed at enhancing the local ocular (mucosal) immune response. PMID- 9875330 TI - Early activation of mitogen-activated protein kinase kinase, extracellular signal regulated kinase, p38 mitogen-activated protein kinase, and c-Jun N-terminal kinase in response to binding of simian immunodeficiency virus to Jurkat T cells expressing CCR5 receptor. AB - We have shown that the binding of simian immunodeficiency virus (SIV) to Jurkat T cells expressing CD4 receptor strongly induces mitogen-activated protein (MAP) kinase kinase (MEK) and extracellular signal-regulated kinases 1 and 2 (ERK1/2) and only weakly induces p38 MAP kinase and c-Jun N-terminal kinase (JNK). Similarly, T-tropic NL4-3 virus, which uses both CD4 and CXCR4 receptors for entry, stimulated in these cells the MEK/ERK MAP kinase (MAPK) pathway in a CD4 receptor-dependent manner (Popik and Pitha, 1998). In contrast, both macrophage tropic SIVmac316 and T cell-tropic SIVmac239, which in addition to CD4 require CCR5 coreceptor for entry, significantly enhanced early MEK/ERK, p38 MAPK, and JNK signaling in Jurkat cells expressing constitutively or transiently the CCR5 receptor. Together, this study provides the evidence that viruses using CXCR4 or CCR5 receptors for entry may differentially use signaling properties of their specific coreceptors to stimulate MAP kinase cascades. In addition, although SIVmac239 and SIVmac316 use different structural domains of the CCR5 receptor for entry, both viruses stimulate early phosphorylation of MEK, ERK, p38, and JNK independently of their tropism and replication. PMID- 9875331 TI - Membrane permeability induced by hepatitis A virus proteins 2B and 2BC and proteolytic processing of HAV 2BC. AB - The ability to rearrange membranes is a unique feature of nonstructural proteins 2B, 2C, and 2BC of some picornaviruses. To analyze in detail membrane binding of the respective proteins of hepatitis A virus (HAV), they were transiently expressed in the vaccinia/T7 system, and their effect on membrane permeability was studied using beta-galactosidase as reporter. Although 2C had no effect, the significantly increased reporter activity observed in the extracellular space of 2B- and 2BC-expressing cells points to a specific effect of HAV proteins 2B and 2BC on membrane permeability. In biochemical fractionation studies, HAV 2C and 2BC showed properties of intregral membrane proteins, whereas 2B was associated with membranes as a peripheral protein. Proteinase 3C-mediated cleavage of precursor 2BC in vivo was most efficient when the enzyme was coexpressed in its precursor forms P3 or 3ABC, which both include the membrane-anchoring domain 3A. 3ABC showed the same solubility pattern as 2BC, suggesting that colocalization of 2BC and 3ABC might be required for the efficient liberation of 2B and 2C and occurs on membranes that have been proposed as the site of viral RNA replication. PMID- 9875332 TI - Gag protein from human immunodeficiency virus type 1 assembles in the absence of cyclophilin A. AB - Human immunodeficiency virus type 1 (HIV-1) replication requires coordinated activities of host and viral factors. We reported previously that interactions of the host factor cyclophilin A with HIV-1 Gag polyproteins affected Gag processing and maturation of virus particles (Streblow et al., 1998. Virology 245, 197-202). We now use in vitro translation and physical analysis of Gag structures to refine our understanding of how cyclophilin A affects HIV-1 replication. Gag assembled into oligomeric structures in vitro in the presence or absence of cyclophilin A, and proteins synthesized under the two conditions were equally susceptible to cleavage by exogenous HIV-1 protease. These and previous data show that Cyclophilin A is required at a step between Gag assembly and Gag processing/virion morphogenesis. Cyclophilin A may be required for Gag conformational changes subsequent to assembly, that are required for efficient dimerization and activation of the viral protease. PMID- 9875333 TI - Binding of the human cytomegalovirus 80-kDa immediate-early protein (IE2) to minor groove A/T-rich sequences bounded by CG dinucleotides is regulated by protein oligomerization and phosphorylation. AB - The 80-kDa immediate-early regulatory protein IE2 of human cytomegalovirus (HCMV) functions as an essential positive transactivator of downstream viral promoters, but it also specifically down-regulates transcription from the major immediate early promoter through a 14-bp DNA target motif known as the cis-repression signal (CRS) located at the transcription start site. The IE2 protein purified from bacteria as a fusion product of either staphylococcal Protein A/IE2(290-579) or glutathione-S-transferase (GST)/IE2(346-579) bound specifically to a [32P] labeled CRS oligonucleotide probe in an in vitro electrophoretic mobility shift assay (EMSA). In contrast, no direct interaction with the CRS probes could be detected with IE2 wild-type protein in extracts from infected or transfected mammalian cells or when synthesized by in vitro translation. However, in vitro phosphorylation of GST/IE2(346-579) by incubation with either the catalytic subunit of protein kinase A (PKA) or a HeLa cell nuclear extract strongly inhibited its DNA-binding activity. This process required ATP hydrolysis and could be reversed by subsequent incubation with bacterial alkaline phosphatase. Importantly, dephosphorylation of the constitutively expressed native IE2 protein present in a nuclear extract from the U373(A45) cell line unmasked a specific CRS DNA-binding activity that could be supershifted with anti-IE2 monoclonal antibody (mAb). A series of high-molecular-weight hetero-oligomeric DNA-bound structures of intermediate mobility were formed in EMSA assays when a mixture of staphylococcal Protein A/IE2 and GST/IE2 was coincubated with the CRS probe. Coincubation with a DNA-binding negative but dimerization-competent GST/IE2 deletion mutant competitively inhibited DNA-binding by staphylococcal Protein A/IE2, whereas coincubation with a GST/IE2 deletion mutant that lacked the ability to both dimerize and bind to DNA failed to influence the mobility of the DNA-bound staphylococcal Protein A/IE2 protein. Therefore, IE2 appears to bind to DNA as a higher-order oligomer in which the presence of subunits with mutant DNA binding domains interferes with the overall DNA-binding function. A series of point mutations introduced into each of nine conserved motifs throughout the DNA binding and dimerization domain, all of which abolish the ability of the transfected intact IE2 protein to autoregulate the MIE promoter, also all lacked the ability to bind to CRS sequences as GST/IE2(346-379) fusion proteins. Detailed analysis of point mutations in the 14-bp CRS target DNA binding motif revealed that IE2 binds in a relatively sequence-independent manner to 10-bp-long A/T-rich DNA elements bounded on each side by CG dinucleotides. Moreover, the A/T rich minor groove binding agent distamycin, but not the G/C-rich minor groove binding agent chromomycin-A3, actively competed with IE2 for binding to the CRS motif in a dose-dependent fashion. In conclusion, IE2 binds preferentially as multimerized dimers to A/T-rich sequences in the minor groove that are flanked on both sides by appropriately spaced CG dinucleotides, and inhibition of the DNA binding or oligomerization activity by PKA phosphorylation probably accounts for the inactivity of the mammalian and in vitro translated forms of the protein. PMID- 9875334 TI - Emergence of epidemic O'nyong-nyong fever in Uganda after a 35-year absence: genetic characterization of the virus. AB - O'nyong-nyong (ONN) virus is an alphavirus (family Togaviridae, genus Alphavirus) classified in the Semliki Forest virus (SFV) antigenic complex. ONN was initially isolated in northern Uganda in 1959 during the early stages of an explosive arbovirus epidemic in which > 2 million cases were reported. No additional epidemics or human isolations of ONN were reported until 1996, when it was isolated from an epidemic in southern Uganda. We report the complete nucleotide and deduced amino acid sequence of one of these 1996-1997 ONN isolates (SG650) and that of the related alphavirus Igbo Ora virus. The data indicate that the recent ONN virus isolate is closely related to the previously published ONN strain isolated in 1959. In addition, phylogenetic analysis of the sequence data reveals that Igbo Ora virus, previously thought to be a separate virus closely related to ONN and Chikungunya (CHIK), clearly is a strain of ONN. The sequence data also reveal that unlike the published ONN (1959) sequence, all ONN strains from the 1996-1997 epidemic possess a stop codon at the nsp3-nsp4 junction. PMID- 9875335 TI - Isolation and stability of histidine-tagged proteins produced in plants via potyvirus gene vectors. AB - A system for the expression and purification of histidine-tagged proteins from plants has been developed using a tobacco etch potyvirus (TEV)-derived gene vectors. The vectors offered a convenient polylinker and a choice of histidine tagging at the recombinant proteins' N or C termini. These vectors were utilized for expression of proteins encoded by beet yellows closterovirus (BYV). Approximately 4 micrograms/g of 20-kDa BYV protein was readily isolated from plants systemically infected by hybrid TEV. In contrast, only minute quantities of 22-kDa BYV capsid protein (CP) histidine-tagged at its N or C terminus could be purified. Rapid degradation of the recombinant CP has been implicated in its failure to accumulate in infected plants. Fusion with TEV HC-Pro stabilized the histidine-tagged BYV CP and facilitated purification of the fusion product from infected plants. This same fusion approach was successfully used with the 24-kDa minor BYV CP. The recombinant proteins were recognized by histidine-tag-specific monoclonal antibody in immunoblot analysis. These results demonstrate the utility of a designed series of TEV vectors for expression, detection, and purification of the recombinant proteins and suggest that intrinsic protein stability is a major factor in a recovery of recombinant proteins from plants. PMID- 9875336 TI - Beta 2-microglobulin knockout mice are highly susceptible to polyoma virus tumorigenesis. AB - Polyoma virus is highly oncogenic when inoculated into immunocompromised adult mice and neonatal mice of specific inbred strains. Although T lymphocytes are known to be essential in controlling polyoma virus tumorigenesis, the importance of class I MHC-restricted CD8+ T cells in mediating tumor resistance remains unclear. Here, we investigated the tumorigenicity of polyoma virus in adult mice rendered CD8+ T cell-deficient by homozygous (-/-) disruption of the beta 2 microglobulin (beta 2m) or CD8 alpha (CD8) genes. Nearly all (94%) of the virus infected adult C57BL/6 beta 2m-/- mice developed tumors, and 20% of the virus inoculated adult C57BL/6CD8-/- mice developed hindlimb paralysis, which is indicative of vertebral tumors. Only 2 of 20 virus-inoculated adult normal C57BL/6 mice developed tumors. Despite these different tumor susceptibilities, persistent viral DNA was detected in multiple organs of mice of all three strains. Multifocal lymphoplasmacytic interstitial infiltrates were present in the kidneys and lungs of virus-infected C57BL/6 beta 2m-/- and in the lungs of virus-inoculated C57BL/6CD8-/- mice. These infiltrates were composed primarily of B cells and colocalized with staining for the major viral capsid protein, VP1. No infiltrates or VP1 staining was detected in the kidneys of infected C57BL/6 mice. Using a highly sensitive RT-PCR bioluminescence immunoassay, we investigated and detected persistent polyoma T protein and VP1 messages in both C57BL/6 beta 2m-/- and C57BL/6 mice. C57BL/6 beta 2m-/- and C57BL/6 mice had equivalent serum virus neutralizing antibody titers. These results provide in vivo evidence that class I MHC-restricted CD8+ T cells are involved in mediating protection against polyoma virus tumor development. PMID- 9875337 TI - Morphological and functional study of dwarf neurons in the rat striatum. AB - Combination of morphological and electrophysiological techniques provided data, suggesting existence in the young rat striatum of a peculiar class of neurons, the neurogliaform or dwarf neurons. Striatal neurons (n = 92), intracellularly recorded from rat brain slices, were filled (one in each slice) with the intracellular marker biocytin, to compare physiological and morphological properties in the same cell. Moreover, some neurons (n = 7) were filled with biocytin plus the fluorescent calcium indicator fura-2, identifying cells during electrophysiological recording. Electrophysiological recording showed that striatal neurons had different firing patterns, suggestive in most cases (n = 80) of spiny neuron class and in others (n = 12) of interneuron class. Fura-2 injection clearly identified the body of six medium-sized cells and of one distinctive tiny cell. This small cell, however, showed a resting membrane potential and spontaneous and evoked firing pattern characteristic of striatal interneurons. Moreover, the fura-2 injected in such small neuron also completely filled the cell body of a near large neuron; the fura-2 fluorescence changed synchronously in the two paired neurons after electrical stimulation of the impaled small one. Accordingly, the biocytin staining identified the morphology of the small recorded neuron as a neurogliaform-like cell apposed to a dendrite of an aspiny neuron, suggesting that the dye injected in one neuron had diffused to the other of a different type. Furthermore, such heterologous dye coupling unexpectedly involved seven pairs of cells detected with biocytin staining (7.6% of the recorded neurons), invariably represented by a medium or large neuron on one side, and on the other side by a small (5.44 +/- 0.15 x 9.14 +/- 0.7 microns, mean +/- SD; n = 7) neurogliaform cell, roundish in shape with few slender and short processes, usually apposed to a dendrite of the companion neurons (six out of seven). In the other cases, the biocytin staining revealed in each slice either the morphology of single spiny or aspiny neurons (80.4% of recorded neurons), or of two-three medium-sized spiny neurons detected near to each other, suggesting that dye coupling had occurred typically between similar neurons (11.9% of the recorded neurons). These data suggest that some neurogliaform cells in the striatum of young rat can be identified as dwarf interneurons, that may be dye-coupled with neurons of different classes. PMID- 9875338 TI - Select types of supporting cell in the inner ear express aquaporin-4 water channel protein. AB - Aquaporins (AQPs) confer a high water permeability on cell membranes and play important parts in secretory and absorptive epithelia in kidney and other organs. Here we investigate whether AQPs are expressed in the sensory epithelia of the inner ear, where a precise volume regulation is crucial. By use of specific antibodies it was found that the inner ear contains AQP1 and 4 while being devoid of detectable levels of AQP2, 3 or 5. Immunofluorescence and postembedding immunogold labelling revealed a strictly non-epithelial distribution of AQP1, confirming previous data. In contrast, AQP4 protein and mRNA (visualized by in situ hybridization) were concentrated in select types of supporting cell, including Hensen's cells and inner sulcus cells. Immunogold particles signalling AQP4 were confined to the basolateral plasma membrane of Hensen's cells and to the basal plasma membrane of Claudius cells and inner sulcus cells. AQP4 was also found in supporting cells of the vestibular end organs, but was absent from transitional epithelial cells and dark cells. Strong labelling for AQP4 and AQP4 mRNA was associated with the central part of the cochlear and vestibular nerves. Hair cells were consistently unlabelled. Our findings indicate that AQP4 may facilitate osmotically driven water fluxes in the sensory epithelia of the inner ear and thus contribute to the volume and ion homeostasis at these sites. PMID- 9875339 TI - Structure and dynamics of receptive fields in the visual cortex of the cat (area 18) and the influence of GABAergic inhibition. AB - Receptive fields (RFs) in the visual cortex are characterized by spatiotemporal profiles that have been described in detail for area 17 simple cells. In this study, we analyse spatial and temporal RF properties of simple and complex cells in layer II/III of area 18 of the anaesthetized adult cat, using the reverse correlation method with brief 50 ms presentations of flashing bright and dark bars. Stimuli were presented with preferred orientation as previously determined by moving bars. Simple cell RFs were characterized by spatially and temporally separable ON and OFF subfields, while in complex cells ON and OFF subfields were superimposed. To discriminate possible contributions of GABAergic inhibition to RF structure and response dynamics in area 18, we have used three-barrelled micropipettes for single cell recordings and microiontophoresis, and have documented ON and OFF responses before, during and after application of bicuculline methiodide for blockade of GABAA receptors. During blockade of GABAergic inhibition, the stimulus-induced and resting discharge frequency increased, and in about 50% of the cells both ON and OFF subfields changed significantly in space and/or time in a reversible manner. In space, blockade of inhibition widened RF subfields, whereas in time, it shortened the duration of the excitatory cell response in simple and complex cells. ON and OFF subfields separated in space and time (simple cells), or time (complex cells) became less isolated or even superimposed. The results indicate substantial local inhibitory processing contributing to spatiotemporal RF properties in layers II/III of area 18 of the cat. PMID- 9875340 TI - Up- and down-expression of the dopamine transporter by plasmid DNA transfer in the rat brain. AB - The functional role of the dopamine transporter (DAT) in central dopaminergic neurotransmission was assessed further by investigating the consequences on dopamine (DA) turn-over of up- and down-regulation of this protein induced by a non-viral gene transfer approach. For this purpose, expression plasmids containing the sense or antisense coding sequence of DAT complexed with the cationic polymer, polyethylenimine (PEI), were injected into the rat substantia nigra, the brain region containing the majority of DA cell bodies. Before in vivo injection, the efficacies of the different DNA constructs were assessed by transfection studies into LLC-PK1 cells. Stereotaxic administration of the sense plasmid complexed to PEI induced, 3 days later, a significant increase in the immunoautoradiographic labelling by anti-DAT antibodies of the substantia nigra and various DA projection areas. These effects were associated with a significantly enhanced capacity of striatal synaptosomes to take up [3H]-DA and lasted up to 14 days postinjection. In contrast, 7 days after intranigral administration of the antisense plasmid complexed to PEI, we observed a significant decrease of DAT immunolabelling in the substantia nigra and [3H]-DA uptake by striatal synaptosomes. Whereas DA turnover in the striatum was unaltered 3 days after intranigral administration of the sense plasmid, it was increased 7 days after intranigral administration of the antisense construct. These data indicate that non-viral transfer of the sense or antisense coding sequence of DAT can be used as a novel approach to induce long-term changes in central DA neurotransmission. PMID- 9875341 TI - Properties of bipolar VIPergic interneurons and their excitation by pyramidal neurons in the rat neocortex. AB - In the rat neocortex, a subset of GABAergic interneurons express the neuropeptide vasoactive intestinal peptide (VIP). Previously, we demonstrated that a population of VIPergic interneurons could be accurately identified by their irregular spiking (IS) pattern and their bipolar morphology. IS interneurons were studied in neocortical slices from 16-22-day-old rats using whole-cell recordings, intracellular labelling and single-cell RT-PCR. In response to a depolarizing pulse, IS interneurons typically discharged a burst of action potentials followed by spikes emitted at an irregular frequency. Several seconds of depolarization, micromolar concentrations of 4-aminopyridine, and nanomolar concentrations of either dendrotoxin I or K converted this irregular pattern to a sustained discharge, suggesting the involvement of an ID-like K+ current. The main glutamate receptor subunits detected in IS cells were GluR1 flop and GluR2 flop, GluR5 and GluR6, and NR2B and NR2D for the alpha-amino-3-hydroxyl-5-methyl 4-isoxazolepropionic acid (AMPA), kainate and N-methyl-D-aspartic acid (NMDA) subtypes, respectively. Paired whole-cell patch-clamp recordings indicated that pyramidal neurons provide intracortical glutamatergic inputs onto IS interneurons. Most connections had high probabilities of response and exhibited frequency-dependent paired pulse depression. Comparison of the amplitude distribution of paired responses suggested that most of these connections consisted of multiple functional release sites. Finally, two discrete subpopulations of IS cells could be identified based on the duration of the initial burst of action potentials and the differential expression of calretinin and choline acetyltransferase. PMID- 9875342 TI - Cloning and characterization of alternative mRNA forms for the rat metabotropic glutamate receptors mGluR7 and mGluR8. AB - Novel mRNA isoforms for two members of the group III metabotropic glutamate receptors (mGluRs), called mGluR7b and mGluR8b, were identified from rat brain cerebral cortex and hippocampus. In both cases, the alternative splicing is generated by a similar out-of-frame insertion in the carboxyl-terminus that results in the replacement of the last 16 amino acids of mGluR7 and mGluR8 by 23 and 16 different amino acids, respectively. Distribution analysis for mGluR7 and mGluR8 isoforms revealed that the two splice variants are generally coexpressed in the same brain areas. The few exceptions were the olfactory bulb, in which only the mGluR7a form could be detected by reverse transcription-polymerase chain reaction, and the lateral reticular and ambiguous nuclei, which showed only mGluR8a labelling. Despite expression in the same regions, different mRNA abundance for the two variants of each receptor were observed. When transiently coexpressed in HEK 293 cells with the phospholipase C-activating chimeric G alpha qi9-G-protein, the a and b forms for both receptor subtypes showed a similar pharmacological profile. The rank order of potencies for both was: DL-amino-4 phosphonobutyrate > L-serine-O-phosphate > glutamate. However, the agonist potencies were significantly higher for mGluR8a, b compared with mGluR7a,b. In Xenopus oocytes, glutamate evoked currents only with mGluR8 when coexpressed with Kir 3.1 and 3.4. Glutamate-induced currents were antagonized by the group II/III antagonist (RS)-alpha-cyclopropyl-4-phosphonophenylglycine. In conclusion, the two isoforms of each receptor have identical pharmacological profiles when expressed in heterologous systems, despite structural differences in the carboxyl terminal domains. PMID- 9875343 TI - Nociceptive integration in the rat spinal cord: role of non-linear membrane properties of deep dorsal horn neurons. AB - Deep dorsal horn neurons (DHNs) involved in nociception can relay long-lasting inputs and generate prolonged afterdischarges believed to enhance the transfer of nociceptive responses to the brain. We addressed the role of neuronal membrane properties in shaping these responses, by recording lamina V DHNs in a slice preparation of the rat cervical spinal cord. Of 256 neurons, 102 produced accelerating discharges in response to depolarizing current pulses, whereas the other neurons showed spike frequency adaptation. Two mechanisms mediated the firing acceleration: a slow inactivation of a K+ current expressed upon activation of the neuron from hyperpolarized holding potentials, and the expression of a regenerative plateau potential activating around resting membrane potential. The increase in firing frequency was much stronger when sustained by the plateau potential (71 DHNs, 28%). A few neurons produced adaptation and both types of acceleration, in different membrane potential domains, showing that the firing pattern of a deep DHN is not a rigid characteristic. Plateau potentials could be elicited by stimulation of nociceptive primary afferent fibres. The bistability associated with plateau potentials permitted afterdischarges. Because plateau potentials had slow activation kinetics and were voltage-dependent, the neurons had non-linear input-output relationships in both the amplitude and time domains. Nociceptive primary afferent stimulation elicited intense and prolonged responses in plateau-generating DHNs, while brief bursts of spikes were evoked otherwise. These results indicate that in a population of deep DHNs, intense firing and prolonged afterdischarges in response to nociceptive stimulation depend on non-linear intrinsic membrane properties. PMID- 9875344 TI - The changing pattern of fibre bundles that pass through the optic chiasm of mice. AB - The organization of retinofugal fibres in the developing and adult mouse has been studied with transmission electron microscopy, autoradiography and the Bodian silver method. It has previously been shown that all retinal ganglion cell axons are in glial-wrapped bundles in the developing and adult optic nerve, but are not in similar bundles close to the chiasm. In the embryonic mouse this region shows a transition in glial morphology from an interfascicular to a radial type and here retinofugal fibres begin to form a new order related to their age. Growth cones become concentrated at the pial surface of the juxtachiasmatic nerve and older fibres are restricted to deeper regions. This same age-related order is also evident in the optic tract. However, the age-related order is lost within the chiasm, where growth cones, young and old fibres are again mingled in distinct bundles as they cross the mid-line. This study is particularly concerned with the structure of the mid-line bundles. These fibre bundles cross each other at right angles, and are recognizable in fetal and adult mice. In the adult, monocular injections of H3 proline followed by autoradiographic study show that the individual mid-line bundles are monocular and that they fuse again, losing the fascicular structure as they leave the chiasm and enter the tract. In the fetus and in the adult, the bundles generally lack a complete glial wrapping so that growth cones can lie in intimate contact with two crossing bundles, one coming from the left eye, the other from the right. The interesting question about the mechanisms that keep growth cones from entering the wrong bundles when they are in this position remains to be addressed. PMID- 9875345 TI - Behavioural characterization of interleukin-6 overexpressing or deficient mice during agonistic encounters. AB - Interleukin-6 (IL-6) is a cytokine released by activated immune cells which has been shown to affect brain function. In this experiment aggressive and affiliative behaviour exhibited during agonistic encounters by transgenic male mice either not expressing (IL-6 -/-) or overexpressing (NSE-hIL-6) IL-6 in the central nervous system was investigated. All subjects were isolated for 24 days before the aggressive encounter and were 52 days old at the time of testing. Subjects were placed for 5 consecutive days in a neutral cage for 15 min with an opponent of the Balb/c strain that had been previously isolated for the same amount of time. The first and the last test sessions were videotaped to evaluate the first approach and the establishment of the social role, respectively. A number of behavioural categories were later scored. When compared with wild-type controls, IL-6 -/- mice showed a higher degree of aggressive behaviour as indicated by a higher frequency of Offensive Upright Posture, an effect more pronounced on the fifth encounter. On the contrary, NSE-hIL-6 subjects showed a tendency to be more involved in affiliative-type social interactions, displaying a higher frequency and duration of behaviours such as Anogenital, Nose or Body Sniff. IL-6 -/- mice showed a clear tendency to exhibit less affiliative interactions compared with their controls while dopamine levels were found to be modified in a number of brain regions in these mice. Overall, these data suggest that IL-6 affects both aggressive and affiliative-type interactions, although the behaviour of the NSE-hIL-6 subjects was less affected than that of the IL-6 -/- group. The effects of the genetic background of the animal in screening the outcome of gene manipulations on behaviour are also discussed. PMID- 9875346 TI - Stimulation of benzodiazepine receptors in the dorsal hippocampus and median raphe reveals differential GABAergic control in two animal tests of anxiety. AB - The effects of pharmacological challenges to the benzodiazepine receptors in the dorsal hippocampus and median raphe nucleus were investigated in the social interaction and the elevated plus-maze tests of anxiety in rats. In the social interaction test, bilateral administration of midazolam (1 and 2 micrograms), into the dorsal hippocampus had anxiolytic effects; flumazenil (500 ng) was silent, but was able to antagonize the anxiolytic effects of midazolam (2 micrograms). In the social interaction test, midazolam was also anxiolytic when infused into the median raphe nucleus; flumazenil (100 and 500 ng) increased locomotor activity, but did not change anxiety measures. As an anatomical control, midazolam (1 and 2 micrograms) was infused into the adjacent pontine reticular nucleus, and was without effect. In contrast to the social interaction test, local infusion of midazolam (1 and 2 micrograms) and flumazenil (100 and 500 ng) into either the dorsal hippocampus or the median raphe nucleus failed to change anxiety measures in the elevated plus-maze (trials 1 and 2). These results show that stimulation of the benzodiazepine receptors in the hippocampus or the median raphe nucleus leads to anxiolytic effects in the social interaction test, but not in the elevated plus-maze. It would therefore appear that the two tests detect different types of anxiety that are differentially modulated by GABAA benzodiazepine receptors in the dorsal hippocampus and the median raphe nucleus. PMID- 9875347 TI - Growth/differentiation factor 5 and glial cell line-derived neurotrophic factor enhance survival and function of dopaminergic grafts in a rat model of Parkinson's disease. AB - Growth/differentiation factor 5 is a member of the transforming growth factor beta superfamily, which has neurotrophic and neuroprotective effects on dopaminergic neurons both in vitro and in vivo. Here we investigate the effects of growth/differentiation factor 5 on foetal mesencephalic grafts transplanted into a rat model of Parkinson's disease, and compare them with those of glial cell line-derived neurotrophic factor. Mesencephalic tissue was suspended in solutions containing either growth/differentiation factor 5 or glial cell line derived neurotrophic factor prior to transplantation into the left striatum of rats with 6-hydroxydopamine lesions of the left medial forebrain bundle. Both proteins enhanced graft-induced compensation of amphetamine-stimulated rotations. Positron emission tomography studies showed that both neurotrophins increased graft-induced recovery of striatal binding of [11C]RTI-121, a marker for dopaminergic nerve terminals. Post mortem analysis at 8 weeks after transplantation showed that both neurotrophins significantly increased the survival of grafted dopaminergic neurons. This study shows that growth/differentiation factor 5 is at least as effective as glial cell line derived neurotrophic factor in enhancing the survival and functional activity of mesencephalic grafts, and thus is an important candidate for use in the treatment of Parkinson's disease. PMID- 9875348 TI - Regions in the human brain activated by simultaneous orientation discrimination: a study with positron emission tomography. AB - In order to compare regional cerebral activity involved in simultaneous as opposed to successive orientation discrimination, we used positron emission tomography to measure regional cerebral blood flow, in two threefold sets of conditions, in a large number of subjects. The first such triad involved simultaneous orientation discrimination, orientation identification and detection, with all tasks using the same pair of gratings. The second triad consisted of successive orientation discrimination with its corresponding identification and detection tasks. Comparisons between tasks within each triad isolate attention to orientation and, respectively, spatial or temporal comparison. The subtraction of detection from simultaneous discrimination revealed activation of right fusiform, right lingual, left precentral, left cingulate and left temporal cortex, in addition to right insula, cerebellum and left thalamus. Only the fusiform, insular and precentral activations remained when the corresponding identification was subtracted from simultaneous discrimination. In contrast, most of the non-visual activation sites remained when simultaneous discrimination was compared with successive discrimination, which also revealed a left lingual activation. These experiments provide further evidence for task-dependent processing in the human visual system and suggest that the right fusiform cortex is involved in spatial as much as temporal comparisons. PMID- 9875349 TI - Immunocytochemical localization of somatostatin receptor sst2A in the rat spinal cord and dorsal root ganglia. AB - Intrathecal administration of octreotide, a stable somatostatin analogue, provides pain relief in patients, and locally applied somatostatin inhibits firing of nociceptive dorsal horn neurons. In the present study, we have raised polyclonal antibodies that specifically detect the somatostatin receptor sst2A and used these antisera for immunocytochemical localization of the receptor protein in the rat spinal cord and dorsal root ganglia. In the superficial layers of the dorsal horn, sst2A-like immunoreactivity (Li) formed a dense network consisting of neuronal perikarya and dendrites which were often closely apposed by, but not co-contained within, somatostatin-14-immunoreactive nerve fibres and terminals. sst2A-Li was resistant to dorsal rhizotomy and did not colocalize with either substance P or calcitonin gene-related peptide suggesting that sst2A-Li was not located to primary afferents, but rather confined to second-order spinal neurons. The position of sst2A-Li perikarya and dendrites in the dorsal horn appeared to be similar to those containing mu-opioid receptor-Li; however, double labelling experiments revealed no instances of coexistence of these two receptors. sst2A-Li was also observed in the dorsal root ganglia predominantly targeted to the somatic plasmalemma of medium size neurons distinct from those expressing somatostatin-14 or delta-opioid receptors. Thus, the present results not only provide a morphological substrate for spinal octreotide analgesia but also show that somatostatin and opioids are poised to modulate nociceptive transmission by distinct anatomical systems. PMID- 9875350 TI - Neurotrophic ACTH4-9 analogue therapy normalizes electroencephalographic alterations in chronic experimental allergic encephalomyelitis. AB - Chronic experimental allergic encephalomyelitis (CEAE) is an established experimental model for multiple sclerosis (MS). The demyelinating lesions in the white matter of the central nervous system observed in CEAE and in MS are accompanied by various neurophysiological alterations. Among the best defined electrophysiological abnormalities are the changes in event-related potentials, in particular evoked potentials involving the spinal cord, i.e. motor and sensory evoked potentials. Less familiar are the changes observed in the electroencephalogram of CEAE-affected animals, which are also encountered in the human equivalent, MS. In the present experiment we evaluated the therapeutic value of a neurotrophic peptide treatment [H-Met(O2)-Glu-His-Phe-D-Lys-Phe-OH, an ACTH4-9 analogue] and its effect on the delayed flash visual evoked potentials (VEP) and power spectra of the electroencephalogram, during a 17-week follow-up of CEAE. CEAE animals treated with the neurotrophic peptide were protected against the development of neurological symptoms during the course of the demyelinating syndrome. VEPs of animals suffering from CEAE showed a delay of the latencies of the late components which was significantly counteracted by peptide treatment. The peak-to-peak amplitude of the VEP afterdischarge recorded from CEAE animals was significantly increased during the course of CEAE and correlated closely with the progression of the myelinopathy. Furthermore, CEAE animals showed an increase of electroencephalogram (EEG) beta activity of up to 500% as compared with the age-matched control group. This increase in beta power mainly consisted of a prevailing 20-21 Hz peak, a frequency that normally is not dominant in control EEG recordings of the rat during passive wakefulness. All these electrophysiological phenomena were absent in ACTH4-9 analogue-treated animals. The present findings underscore the potential importance of a neurotrophic peptide treatment in the pharmacotherapy of central demyelinating syndromes, and possibly of MS. PMID- 9875351 TI - Subcellular and subsynaptic distribution of the NR1 subunit of the NMDA receptor in the neostriatum and globus pallidus of the rat: co-localization at synapses with the GluR2/3 subunit of the AMPA receptor. AB - Glutamatergic neurotransmission in the neostriatum and the globus pallidus is mediated through NMDA-type as well as other glutamate receptors and is critical in the expression of basal ganglia function. In order to characterize the cellular, subcellular and subsynaptic localization of NMDA receptors in the neostriatum and globus pallidus, multiple immunocytochemical techniques were applied using antibodies that recognize the NR1 subunit of the NMDA receptor. In order to determine the spatial relationship between NMDA receptors and AMPA receptors, double labelling was performed with the NR1 antibodies and an antibody that recognizes the GluR2 and 3 subunits of the AMPA receptor. In the neostriatum all neurons with characteristics of spiny projection neurons, some interneurons and many dendrites and spines were immunoreactive for NR1. In the globus pallidus most perikarya and many dendritic processes were immunopositive. Immunogold methods revealed that most NR1 labelling is associated with asymmetrical synapses and, like the labelling for GluR2/3, is evenly spread across the synapse. Double immunolabelling revealed that in neostriatum, over 80% of NR1-positive axospinous synapses are also positive for GluR2/3. In the globus pallidus most NR1-positive synapses are positive for GluR2/3. In both regions many synapses labelled only for GluR2/3 were also detected. These results, together with previous data, suggest that NMDA and AMPA receptor subunits are expressed by the same neurons in the neostriatum and globus pallidus and that NMDA and AMPA receptors are, at least in part, colocalized at individual asymmetrical synapses. The synaptic responses to glutamate in these regions are thus likely be mediated by both AMPA and NMDA receptors at the level of individual synapses. PMID- 9875352 TI - Direct glutamate-mediated presynaptic inhibition of sensory afferents by the postsynaptic motor neurons. AB - An in vitro preparation of the crayfish central nervous system was used to study a negative feedback control exerted by the glutamatergic motor neurons (MNs) on to their presynaptic cholinergic sensory afferents. This negative control consists in small amplitude, slowly developing depolarizations of the primary afferents (sdPADs) strictly timed with MN bursts. They were not blocked by picrotoxin, but were sensitive to glutamate non-N-methyl-D-aspartate (NMDA) antagonists. Intracellular recordings were performed within thin branches of sensory terminals while electrical antidromic stimulation were applied to the motor nerves, or while glutamate (the MN neurotransmitter) was pressure-applied close to the recording site. Electrical motor nerve stimulations and glutamate pressure application had similar effects on to sensory terminals issued from the coxo-basipodite chordotonal organ (CBTs): like sdPADs, both stimulation-induced depolarizations were picrotoxin-resistant and were dramatically reduced by non NMDA antagonist bath application. These results indicate that sdPADs are likely directly produced by MNs during locomotor activity. A functional scheme is proposed. PMID- 9875353 TI - Differential time- and dose-related effects of haemorrhage on tyrosine hydroxylase and neuropeptide Y mRNA expression in medullary catecholamine neurons. AB - Hypotensive haemorrhage induces nuclear Fos expression and upregulates tyrosine hydroxylase (TH) mRNA in catecholamine-containing cell groups of the rat medulla oblongata. To shed light on the significance of the coexistence of neuropeptide Y (NPY) in aminergic neurons, the impact of graded levels of haemorrhage on temporal changes in the expression of TH and NPY mRNAs was compared; concurrent staining for Fos permitted comparisons between cells that ostensibly were and were not targeted by the stimulus. A 15% haemorrhage provoked increased NPY expression in all medullary catecholamine cell groups except the A2; these changes were detected predominantly in Fos-immunoreactive neurons (Fos-ir) at later (2-4 h) time points. Upregulation of TH and NPY mRNAs in Fos-ir neurons followed distinct time courses, with NPY responses peaking more rapidly, particularly in the C1 and C2 cell groups. Adrenergic cell groups displayed greater maximal increases in NPY expression than the A1 noradrenergic cell group while the converse was true of TH mRNA response. Increasing the severity of haemorrhage resulted in more pronounced increases in both mRNA responses in each aminergic region. These findings indicate that haemorrhage differentially affects TH and NPY expression in medullary catecholamine cell groups that participate in the maintenance of cardiovascular homeostasis. The differential nature of these responses suggests them not to be a simple consequence of metabolic alterations pursuant to increased synaptic activity. The prompt and robust NPY mRNA responses in adrenergic neurons suggests a mechanism by which peptide content of these cell groups' terminal projections is defended. PMID- 9875354 TI - Continuous blockade of brain glucocorticoid receptors facilitates spatial learning and memory in rats. AB - Previously, a corticosterone surge associated with a learning task was shown to facilitate cognitive processes through brain glucocorticoid receptors (GR) while chronic overexposure to this stress hormone impaired cognition. In the present study we tested the hypothesis that opposing effects on learning and memory might also occur after either phasic or continuous blockade of brain GR by intracerebroventricular (i.c.v.) administration of the GR antagonist RU38486 (aGR). We used a Morris water maze procedure to assess spatial learning and memory abilities in male Wistar rats. The effect of phasic brain GR blockade was studied following daily pretraining administration of 10 and 100 ng/microL aGR i.c.v. on 3 consecutive days. This repetitive aGR treatment impaired spatial learning and memory dose-dependently in comparison with vehicle controls. For continuous brain GR blockade, animals received an i.c.v., infusion of aGR (10 and 100 ng/0.5 microL per h or vehicle) over 10 days. Infusion of 100 ng aGR per hour resulted in a long-lasting facilitation of spatial performance. The 10 ng aGR infusion also caused initially a facilitating effect, which was, however, transient and performance became impaired during retest. Possible anxiolytic properties of the drugs were excluded in view of the animals' behaviour in the elevated plus maze. Both doses of aGR infusion reduced the number of mineralocorticoid receptors in the hippocampus, but only the high dose of aGR resulted in a significant reduction of available GR sites. In conclusion, continuous administration of GR antagonist improves cognitive function, while phasic blockade of brain GR function causes a cognitive deficit. PMID- 9875355 TI - Direction discrimination of moving gratings and plaids and coherence in dot displays without primary visual cortex (V1). AB - We present new experimental observations of G.Y., a well-tested patient with unilateral loss of primary visual cortex. We stimulated G.Y.'s blind hemifield using first- and second-order motion stimuli at velocities around psychophysical threshold. Using a dual response paradigm (awareness level of visual motion, motion direction discrimination) psychophysical performance improved with increasing velocity and dot coherence. We were also able to influence directly G.Y.'s performance for the better and at will, by placing the emphasis solely on direction discrimination. In the absence of V1, graduated detection and discrimination of stimuli known to activate both V1 and extrastriate motion areas MT/V5 and MST is still possible. These results are in line with residual visual processing but did not show evidence of unconscious processing of motion stimuli characteristic of 'blindsight'. PMID- 9875356 TI - Transient features of the thalamic reticular nucleus in the human foetal brain. AB - The architectonic organization and neuronal types of the human foetal reticular nucleus (RN)--with special reference to transient characteristics--have been investigated using antisera against calretinin, parvalbumin and neurofilament epitopes of somata and dendrites (SMI 311). The RN consists of four subdivisions (clearly distinguishable in the 6/7th gestational month): The main portion appears as a prominent structure on account of its extension and high packing density of neurons which coexpress calretinin and parvalbumin. These two calcium binding proteins are also expressed by the perireticular nucleus forming a conspicuous grey within the internal capsule. Perireticular cells form clusters which are in continuity with the main portion, globus pallidus, ganglionic eminence and pregeniculate nucleus. In double-labellings, a medial subnucleus stands out distinctly as it only expresses calretinin. SMI 311-immunopreparations show neurons revealing a high degree of diversification and elaborated dendritic trees. Several transient characteristics become obvious: the perireticular nucleus, not visible in the adult, represents a distinct entity in the human foetal brain. The main portion and the pregeniculate nucleus appearing as prominent greys are dramatically reduced in size later on. The percentage of RN neurons expressing calretinin, the diversity of neuronal types and elaborated dendritic trees are reduced. The transient features can be correlated with the RN's putative functional roles in development: early RN-afferents to the dorsal thalamus may represent pioneer fibres providing guiding cues for outgrowing axons from or into the thalamus. Moreover, the RN may serve as an intermediate target for growing axons which are sorted and directed towards different final targets. PMID- 9875357 TI - beta-Actin is confined to structures having high capacity of remodelling in developing and adult rat cerebellum. AB - Neurons undergo complex morphological changes during differentiation and in cases of plasticity. A major determinant of cell morphology is the actin cytoskeleton, which in neurons is comprised of two actin isoforms, non-muscle gamma- and beta actin. To better understand their respective roles during differentiation and plasticity, their cellular and subcellular localization was examined in developing and adult cerebellar cortex. It was observed that gamma-actin is expressed at a constant level throughout development, while the level of beta actin expression rapidly decreases with age. At the light microscopic level, gamma-actin staining is ubiquitous and the only developmental change observed is a relative reduction of its concentration in cell bodies and white matter. In contrast, beta-actin staining almost completely disappears from the cytoplasm of cell bodies, primary dendrites and axons. In young cerebellar cultures, gamma actin is found in the cell body, neurites and growth cones, while beta-actin is mainly found in growth cones, as previously reported in other primary neuronal culture systems [Kaech et al. (1997), J. Neuroscience, 17, 9565-9572; Bassell et al., (1998), J. Neuroscience, 18, 251-265]. Electron microscopy of post-embedding immunogold-labelled tissue confirms the widespread distribution of gamma-actin, and also reveals an increased concentration of gamma-actin in dendritic spines in the adult. During development, beta-actin accumulation is observed in actively growing structures, e.g., growth cones, filopodia, cell bodies and axonal tracts. In the adult cerebellar cortex, beta-actin is preferentially found in dendritic spines, structures which are known to retain their capacity for morphological modifications in the adult brain. This differential subcellular localization and developmental regulation of the two actin isoforms point to their different roles in neurons. PMID- 9875358 TI - Development of the histaminergic neurons and expression of histidine decarboxylase mRNA in the zebrafish brain in the absence of all peripheral histaminergic systems. AB - The histamine-storing neural system in adult and developing zebrafish (Danio rerio) was studied with immunocytochemical and chromatographical methods. Furthermore, the gene for histidine decarboxylase was partially cloned and its expression mapped with in situ hybridization. The histamine-storing neurons were only seen in the caudal hypothalamus, around the posterior recess of the diencephalic ventricle. Almost all parts of the brain, except the cerebellum, contained at least some histamine-immunoreactive fibres. The ascending projections had the rostral part of the dorsal telencephalon as a major target. Descending projections terminated in the torus semicircularis, central grey and inferior olive. A prominent innervation of the optic tectum, which has not been reported in other fish, was seen. The in situ hybridization gave a strong signal in cells with the same anatomical position as the histamine-immunoreactive neurons. The first histamine-immunoreactive neurons appeared in the ventral hypothalamus at about 85 h post-fertilization, and at 90 h, immunoreactive fibres terminated in the dorsal telencephalon. The embryonic histamine production described in mammals was lacking in this species. Both immunocytochemical and chromatographical studies indicated that histamine is absent in all other parts of the zebrafish body, and no specific hybridization was seen in any other part of the fish than the hypothalamus. The zebrafish could therefore be a very useful model for pharmacological in vivo studies of the histaminergic system of the brain, since the powerful peripheral actions of histamine should be lacking in this species. PMID- 9875359 TI - Stratum radiatum giant cells: a type of principal cell in the rat hippocampus. AB - Neurons of a distinct type in CA1 area stratum radiatum of the rat hippocampus have been found to express a direct cellular form of long-term potentiation (LTP, Maccaferri & McBain, 1996, J. Neurosci. 16, 5334), but their functional identity, i.e. whether interneuron or principal cell, remained unknown. Whole cell recording from hippocampal slices in vitro was combined with light and electron microscopy to answer this question. LTP was robustly induced by a pairing protocol and physiological properties were measured in radiatum giant cells (RGCs) using biocytin containing pipettes. Reconstruction of the cells' dendritic and axonal arbor revealed morphological properties similar to CA1 pyramidal cells with some characteristic differences. They typically had two large diameter apical dendrites, or when only one dendrite arose, it soon bifurcated. Apical dendrites formed a dendritic tuft in stratum lacunosum-moleculare and the dendrites, but not the somata, were densely covered with conventional spines. The axon arose from the basal pole of the soma, descended to stratum oriens and emitted several axon terminals bearing collaterals that travelled horizontally, remaining in stratum oriens. The main, myelinated axon trunks turned towards the fimbria. In the electron microscope axon terminals were found to form asymmetrical synapses on postsynaptic dendritic shafts and dendritic spines in stratum oriens. The dendrites received asymmetrical synapses, mostly on their spines. The axon initial segments also received several synapses, a feature never observed on interneurons. All the above characteristics support the conclusion that RGCs are excitatory principal neurons. PMID- 9875360 TI - Synaptic inhibition in the isolated respiratory network of neonatal rats. AB - Gramicidin-perforated patch-clamp recording revealed phasic Cl(-)-mediated hyperpolarizations in respiratory neurons of the brainstem-spinal cord preparation from newborn rats. The in vitro respiratory rhythm persisted after block of gamma-aminobutyric acid (GABA), i.e. GABAA, receptor-mediated inhibitory postsynaptic potentials (IPSPs) with bicuculline and/or glycinergic IPSPs with strychnine. In one class of expiratory neurons, bicuculline unmasked inspiration related excitatory postsynaptic potentials (EPSPs), leading to spike discharge. Bicuculline also blocked hyperpolarizations and respiratory arrest due to bath applied muscimol, whereas strychnine antagonized similar responses to glycine. The reversal potential of respiration-related IPSPs and responses to GABA, muscimol or glycine was not affected by CO2/HCO3(-)-free solutions, but shifted from about -65 mV to values more positive than -20 mV upon dialysis of the cells with 144 instead of 4 mM Cl-. Impairment of GABA uptake with nipecotic acid or glycine uptake with sarcosine evoked a bicuculline- or strychnine-sensitive decrease of respiratory frequency which could lead to respiratory arrest. Also, the GABAB receptor agonist baclofen led to reversible suppression of respiratory rhythm. This in vitro apnoea was accompanied by a K+ channel-mediated hyperpolarization (reversal potential -88 mV) of tonic cells, whereas membrane potential of neighbouring respiratory neurons remained almost unaffected. Both baclofen-induced hyperpolarization and respiratory depression were antagonised by 2-OH-saclofen, which did not affect respiration-related IPSPs per se. The results show that synaptic inhibition is not essential for rhythmogenesis in the isolated neonatal respiratory network, although (endogenous) GABA and glycine have a strong modulatory action. Hyperpolarizing IPSPs mediated by GABAA and glycine receptors provide a characteristic pattern of membrane potential oscillations in respiratory neurons, whereas GABAB receptors rather appear to be a feature of non respiratory neurons, possibly providing excitatory drive to the network. PMID- 9875361 TI - Acute phencyclidine neurotoxicity in rat forebrain: induction of haem oxygenase-1 and attenuation by the antioxidant dimethylthiourea. AB - Phencyclidine and other N-methyl-D-aspartate receptor antagonists are toxic to pyramidal neurons in the posterior cingulate/retrosplenial cortex of rat brain. Previous studies have shown induction of heat shock protein 70 in affected neurons. In this study, expression of haem oxygenase-1, a heat shock protein induced by oxidative stress, was examined in rat forebrain after administration of a single intraperitoneal dose of phencyclidine (50 mg/kg). Northern and Western blot analyses of brain tissue extracts from phencyclidine-treated rats revealed a marked induction of haem oxygenase-1 mRNA and protein, respectively. Immunohistochemistry studies revealed that phencyclidine increased haem oxygenase 1 immunoreactivity primarily in posterior cingulate/retrosplenial, piriform and entorhinal cortices, striatum and hippocampus. Haem oxygenase-1 protein was induced in non-neuronal cells, mainly astrocytes. Some microglia expressing haem oxygenase-1 protein were also found in the posterior cingulate/retrosplenial cortex. Haem oxygenase-1 immunoreactive astrocytes and microglia were present in close proximity to the heat shock protein 70-positive neurons in the posterior cingulate/retrosplenial cortex following phencyclidine. Pretreatment of rats with 1,3-dimethylthiourea, an antioxidant, significantly reduced haem oxygenase-1 protein induction by phencyclidine. Thus, induction of haem oxygenase-1 in glia by phencyclidine appears to be mediated mostly by oxidative stress. Experiments with the amino cupric silver stain for neuronal degeneration revealed phencyclidine-induced neurotoxicity in the posterior cingulate/retrosplenial cortex. The number of affected neurons was significantly reduced after 1,3 dimethylthiourea pretreatment. This suggests that the neurotoxicity of N-methyl-D aspartate antagonists is due in part to the oxidative stress and may be amenable to therapeutic interventions. PMID- 9875362 TI - Proliferation, migration and differentiation of neuronal progenitor cells in the adult mouse subventricular zone surgically separated from its olfactory bulb. AB - The subventricular zone of the adult mammalian forebrain contains progenitor cells that, by migrating along a restricted pathway called the 'rostral migratory stream' (RMS), add new neurons to the olfactory bulb throughout life. To determine the influence of the olfactory bulb on the development of these progenitor cells, we performed lesions that interrupt this pathway and separate the olfactory bulb from the rest of the forebrain. By labelling cells born at several survival times after the lesions with the thymidine analogue bromodeoxyuridine (BrdU), we found that disconnection from the bulb influences the rate of BrdU incorporation by the progenitor cells. The number of labelled cells in lesioned mice was almost half that found in control mice. In the disconnected migratory pathway, the number of neurons expressing calretinin was increased indicating that neuronal differentiation was enhanced: newly born neurons occurred within and around the RMS, most of them expressed calretinin and left the pathway starting about 2 weeks after the lesion. Thereafter, these neurons preserving their phenotype, spread for long distances, and accumulated ectopically in dorsal regions of the anterior olfactory nucleus and the frontal cortex. Finally, transplantation of adult subventricular cells into the lesioned pathway showed that the lesion neither prevents neuronal migration nor alters its direction. Thus, although the olfactory bulb appears to regulate the pace of the developmental processes, its disconnection does not prevent the proliferation, migration and phenotypic acquisition of newly generated bulbar interneurons that, since they cannot reach their terminal domains, populate some precise regions of the lesioned adult forebrain. PMID- 9875363 TI - Functional relationship between age-related immunodeficiency and learning deterioration. AB - Disordered immune responses are supposed to alter the function of the central nervous system through the neuroendocrine immunomodulation network. In this paper, we studied the influence of the immune function on learning performances from the angle of pharmacology using aged garlic extract (AGE), an immunomodulator. Splenocyte proliferation, induced by concanavalin A or lipopolysaccharide, and the antibody production response were declined in senescence accelerated mouse-prone 8 (SAMP8) aged 12 months compared with age matched SAMR1 (SAM-resistant 1). Chronic oral administration of AGE-containing food (2%, w/w) significantly enhanced the immune responses of both SAMP8 and SAMR1. Male ddY mice were thymectomized 4 weeks after birth and fed AGE containing food after the operation until the experiments were finished. Learning performances, brain monoamine content and choline acetyltransferase (ChAT) activity, as well as the immune response were evaluated 10 months after the operation. Thymectomy resulted in not only immunodeficiency, but also deteriorated learning ability. AGE treatment prevented the reduction of the antibody production response induced by thymectomy and improved the thymectomy induced deterioration of learning behaviours in passive avoidance performance and in a spatial memory task. The contents of hypothalamic noradrenaline, 3,4 dihydroxyphenylacetic acid and homovanillic acid, and the hypothalamic ChAT activity were increased in thymectomized mice compared to those of sham-operated control, while AGE treatment restored them to the control levels. These results suggest that the improvement of immune function is closely related to the amelioration of age-associated deterioration of learning and memory. PMID- 9875364 TI - Different regions of the periaqueductal grey are involved differently in the expression and conditioned inhibition of fear-potentiated startle. AB - The fear-potentiated startle paradigm is a useful model for investigating the neural systems underlying the mediation and modulation of fear and anxiety. A series of previous studies showed that the periaqueductal grey is involved in the expression and inhibition of fear-potentiated startle, but it was uncertain whether the different columns of the periaqueductal grey play different roles. In the present study, picrotoxin, a GABAA (gamma-aminobutyric acid) chloride-channel blocker, was injected into the dorsal, lateral and ventrolateral columns of the periaqueductal grey in order to examine the effects of neural disinhibition on expression and conditioned inhibition of fear-potentiated startle. Picrotoxin injections into the lateral periaqueductal grey increase potentiation of the startle response by conditioned fear, injections into the ventrolateral periaqueductal grey block fear-potentiated startle. Picrotoxin injections into the dorsal periaqueductal grey decreased conditioned inhibition but did not affect expression of fear-potentiated startle. The baseline startle amplitude was not influenced by picrotoxin injections into the different columns of the periaqueductal grey. Picrotoxin injections into neighbouring brain areas did not have any effect on baseline startle amplitude nor did they have an effect on expression or conditioned inhibition of fear-potentiated startle. Based on the present results, the possible pathways underlying the expression and conditioned inhibition of fear-potentiated startle in rats are discussed. PMID- 9875365 TI - Shaping of the cat paw for food taking and object manipulation: an X-ray analysis. AB - The kinematics of the cat distal forelimb during food-taking were analysed to obtain information on the movement processes within the paw before and during object taking in a species without monosynaptic corticomotoneuronal projections. The behaviour was investigated with two tests: either the table test (TT, food offered on a table located at ground level in a reaching distance of 22 and 28 cm) or the horizontal test (HT, food offered in a small container located at shoulder level, height 18-25 cm, reaching distance 6-12 cm). In five animals, the changes in configuration and the conjoint actions of the wrist, the metacarpophalangeal (MCP) and proximal interphalangeal (PIP) joints were assessed with three-dimensional X-ray cinematography (time resolution 20 ms, spatial resolution 1 mm) and video analysis. While approaching the target, the digits were first extended and subsequently abducted. This 'preshaping' consisted of combined angular changes in the MCP and PIP joints, thereby attaining an adequate grip aperture. Each cat used a stable strategy, but different cats used different strategies. In the TT, preshaping involved an MCP extension and a PIP flexion. In the HT, predominant extension of the MCP, predominant extension of the PIP, or a combination of both were used, followed by MCP flexion and PIP flexion. The grip aperture started to decrease before object contact, earlier in the TT, later in the HT. Grasping was achieved by flexion of first the PIP and later the MCP. The X-ray analysis gave evidence for individuated digit movements. Correlation analysis of the angular position of the joints between the different phalanges showed that digits 3 and 4 acted in concert, as did digits 2 and 5, but with clear independence between the different pairs. Furthermore, the different phalanges served different purposes during the grasp. Digits 3, 4 established object contact, digits 2, 5 were mainly used to stabilize the paw onto the surface. The cat distal forelimb displays a refined movement repertoire during the preshaping and grasping phase of food-taking. During the preshaping phase, the kinematics resembled in some aspects the situation in humans. The results demonstrate the ability of the polysynaptic projections from the cortico motoneuronal system to organize differentiated distal limb movements, including individuated movements of the digits. PMID- 9875366 TI - A purinergic component of the excitatory postsynaptic current mediated by P2X receptors in the CA1 neurons of the rat hippocampus. AB - The pyramidal neurons in the CA1 area of hippocampal slices from 17- to 19-day old rats have been investigated by means of patch clamp. Excitatory postsynaptic currents (EPSCs) were elicited by stimulating the Schaffer collateral at a frequency below 0.2 Hz. It was found that inhibition of glutamatergic transmission by 20 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 100 microM 2-amino-5-phosphonovaleric acid (D-APV) left a small component of the EPSC uninhibited. The amplitude of this residual EPSC (rEPSC) comprised 25 +/- 11% of the total EPSC when measured at a holding potential of -50 mV. The rEPSC was blocked by selective P2 blocker pyridoxal phosphate-6-azophenyl-2'-4'-disulphonic acid (PPADS) 10 microM and bath incubation with non-hydrolysable ATP analogues, ATP-gamma-S and alpha, beta-methylene-ATP at 50 and 20 microM, respectively. The rEPSC was dramatically potentiated by external Zn2+ (10 microM). In another series of experiments exogenous ATP was applied to the CA1 neurons in situ. An inward current evoked by ATP was inhibited by PPADS to the same extent as the rEPSC. It is concluded that, depending on membrane voltage, about one-fifth to one-quarter of the EPSC generated by the excitatory synaptic input to the hippocampal CA1 neurons of rat is due to the activity of P2X receptors. PMID- 9875367 TI - Individual differences in stress-induced dopamine release in the nucleus accumbens are influenced by corticosterone. AB - Stressful experiences, glucocorticoids hormones and dopaminergic neurons seems to interact in determining a higher propensity to develop drug abuse. In this report, we studied the acute interaction between these three factors. For this purpose, we compared stress-induced dopamine release in intact rats and in rats in which stress-induced corticosterone secretion was experimentally blocked. Ten minute tail-pinch was used as a stressor and dopamine release estimated in the nucleus accumbens by using the microdialysis technique. Individual differences were also taken into account by comparing rats identified as either predisposed (HRs) or resistant (LRs) to develop self-administration of drugs of abuse, on the basis of their locomotor response to novelty. It was found that suppression of stress-induced corticosterone secretion significantly decreased stress-induced dopamine release. However, such an effect greatly differed between HR and LR rats. When corticosterone secretion was intact HR animals had a higher and longer dopamine release in response to stress than LRs. The blockade of stress-induced corticosterone secretion selectively reduced the dopaminergic response of HRs that did not differ from LRs anymore. These findings strength the idea that glucocorticoids could be involved in determining propensity to develop drug self administration. In particular, these hormones could play a role in determining the higher dopaminergic activity that characterizes drug proned individuals. PMID- 9875368 TI - Functional expression of GFP-tagged Kv1.3 and Kv1.4 channels in HEK 293 cells. AB - Various types of voltage gated potassium channels (Kv) are responsible for setting the resting potential and shaping the membrane potential waveform in the subcellular domains of neurons. In order to visualize the expression behaviour of recombinant Kv channels, we have fused green fluorescent protein (GFP) to the N terminal of the alpha subunits Kv1.3 and Kv1.4. In transiently transfected HEK 293 cells the GFP-Kv chimeras localize to the plasma membrane. Whole-cell voltage clamp recordings demonstrate that they form functional potassium channels. Kinetic analysis reveals that the gating kinetics of GFP-Kv1.3 are virtually indistinguishable from those displayed by its wild-type correlate. For GFP-Kv1.4 channels we find that their gating is modified in an expected manner. In response to short depolarizing voltage pulses they do not inactivate, indicating that the attached GFP interferes with the fast N-type inactivation mechanism present in wild type Kv1.4 channels. We suggest that GFP tagging of Kv channels might be a useful tool to monitor the spatiotemporal distribution of recombinant potassium channels expressed in living neurons. PMID- 9875369 TI - Hippocampal synaptic density and glutamate immunoreactivity following transient cerebral ischaemia in the chick. AB - A transient ischaemic episode of 10 min duration was induced in 1-day-old chicks. After a 1-week survival period, synapse density was assessed in the ventral hippocampus using the 'disector' technique. A significant decrease was observed in asymmetric synapses, markedly greater than that observed previously in the dorsal hippocampus. Because the effect occurred mainly on excitatory synapses, the distribution of glutamate in the ventral hippocampus was also assessed by a postembedding immunogold labelling technique. The density of gold particles was significantly greater in both boutons and neuropil in the ischaemic group compared to controls, lending support to the theory of excitotoxicity as an explanation for ischaemic neural degeneration. PMID- 9875370 TI - How does the human brain deal with a spinal cord injury? AB - The primary sensorimotor cortex of the adult brain is capable of significant reorganization of topographic maps after deafferentation and de-efferentation. Here we show that patients with spinal cord injury exhibit extensive changes in the activation of cortical and subcortical brain areas during hand movements, irrespective of normal (paraplegic) or impaired (tetraplegic patients) hand function. Positron emission tomography ([15O]-H2O-PET) revealed not only an expansion of the cortical 'hand area' towards the cortical 'leg area', but also an enhanced bilateral activation of the thalamus and cerebellum. The areas of the brain which were activated were qualitatively the same in both paraplegic and tetraplegic patients, but differed quantitatively as a function of the level of their spinal cord injury. We postulate that the changes in brain activation following spinal cord injury may reflect an adaptation of hand movement to a new body reference scheme secondary to a reduced and altered spino-thalamic and spino cerebellar input. PMID- 9875371 TI - The design, synthesis and properties of highly potent and selective inhibitors of herpes simplex virus types 1 and 2 thymidine kinase. AB - The rational design and synthesis of nucleotide analogues as inhibitors of herpes simplex virus (HSV) thymidine kinase is described. Starting from thymidine, product analogues which included phosphates, phosphonates, sulphonates, sulphonamides and carboxamides were prepared. The carboxamide series showed good structure-activity relationships and afforded a lead structure which inhibited the HSV-2 enzyme in the low micromolar range. Replacing the 5-methyl group in thymidine by ethyl enhanced the potency of the lead structure 10-fold. Further optimization of the carboxamide moiety afforded inhibitors active in the sub nanomolar range and finally the introduction of a 2'-beta-fluoro substituent improved the potency a further twofold. The low water solubility of the most potent inhibitor was overcome by conversion to the 3'-valyl ester, which had good oral bioavailability and showed activity by the oral route in murine models of infection. PMID- 9875372 TI - Inhibition of virus-encoded thymidine kinase suppresses herpes simplex virus replication in vitro and in vivo. AB - Both herpes simplex virus type 1 (HSV-1) and HSV-2 encode a thymidine kinase enzyme which differs from cellular thymidine kinase in substrate specificity. Viral thymidine kinase enables the virus to replicate in cells that lack cellular thymidine kinase, namely those of the sensory neurons where the virus establishes, and periodically reactivates from, a latent state. Thymidine kinase dependent HSV replication following viral reactivation at the site of latency is thought to precede the emergence of virus at mucosal surfaces. The ability to inhibit such an essential viral enzyme would potentially prevent HSV from replicating within neuronal tissue, and thus stop the recurrent disease cycle. Ro 32-2313 was designed as a selective and competitive inhibitor of HSV thymidine kinase and in vitro studies have confirmed this mechanism of action. In vivo evaluation of a soluble prodrug of Ro 32-2313, Ro 32-4397, was undertaken in murine models where pathogenesis was dependent upon viral replication in neuronal tissue. It was shown that in vivo administration of Ro 32-4397 (i) significantly reduced the viral titre detected in isolated dorsal root ganglia; (ii) prevented HSV-2-induced lethality in a systemic infection model; and (iii) reduced zosteriform lesion development in a model of dermal infection. Administration of Ro 32-4397 produced dose-related changes in viral pathogenicity towards those of the phenotype of a thymidine kinase-deficient virus. Overall, the study confirmed that thymidine kinase inhibitors can suppress the replication of HSV in vivo, and suggest that such inhibitors may reduce reactivation of the virus from latency if used prophylactically in recurrent HSV infection. PMID- 9875373 TI - Combinations of antiviral and anti-inflammatory preparations for the topical treatment of herpes simplex virus assessed using a murine zosteriform infection model. AB - Recently we have reported a zosteriform murine infection model which employs the adoptive transfer of immune cells (ATI) to recipient infected mice to produce a disease that mimics human recurrent herpes simplex virus (HSV) disease. Mice were infected with HSV-1 by scarification at the lateroventral line of the neck; 2 days later, the mice received immune cells from HSV-1-infected syngeneic mice. Although virus was cleared more quickly from the target tissues of virus replication in recipient mice, ATI resulted in a heightened inflammatory response and delayed healing. This model was used to test the effects of topical formulations containing foscarnet and/or the anti-inflammatory agent, hydrocortisone. Virus clearance and clinical signs, including ear thickness and zosteriform spread of lesions, were studied. Treatment with 3% foscarnet accelerated virus clearance but had little effect on clinical parameters. By contrast, 0.5% hydrocortisone increased the titre and extended the presence of infectious virus for at least 6 days, although the reduction in clinical signs was greater than that obtained with topical foscarnet. Foscarnet in combination with hydrocortisone produced a marked reduction in clinical signs while virus replication was reduced. These results are discussed in relation to the inflammation and discomfort experienced by patients and a possible role for anti inflammatory formulations in the treatment of HSV reactivation episodes in man. PMID- 9875374 TI - Enzymatic synthesis of 2'-O-acyl prodrugs of 1-(beta-D-arabinofuranosyl)-5(E)-(2 bromovinyl)uracil and of 2'-O-acyl-araU, -araC and -araA. AB - Pig liver esterase (EC 3.1.1.1) catalysed regioselective hydrolysis of 1-(2,3,5 tri-O-acyl-beta-D-arabinofuranosyl)uracil, -cytosine and -adenine to give the corresponding 2'-monoesters effectively and in high yield. This methodology enabled the preparation of 1-(2-O-acyl-beta-D-arabinofuranosyl)-5-[(E)-(2 bromovinyl)]uracil prodrugs which, although slightly less active than the parent 1-(beta-D-arabinofuranosyl)-5-(E)-(2 bromovinyl)uracil (sorivudine; BV-araU), were strongly active in vitro against varicella-zoster virus (ED50 2.4-45 ng/ml). The retarded rates of enzymatic hydrolysis of the 2'-esters imply that they might function as lipophilic prodrugs, leading to increased plasma and cellular concentrations. In view of the marked in vitro activity, they represent an interesting approach to arabinofuranosyl nucleoside prodrugs with improved pharmacokinetics and enzymatic stability. PMID- 9875375 TI - Alkylthioglycerol prodrugs of foscarnet: synthesis, oral bioavailability and structure-activity studies in human cytomegalovirus-, herpes simplex virus type 1 and human immunodeficiency virus type 1-infected cells. AB - In a previous study, we reported that 1-O-octadecyl-sn-glycero-3-foscarnet (ODG PFA) was 40 to 93 times more potent than free foscarnet (PFA) in human cytomegalovirus (HCMV)-, herpes simplex virus type 1 (HSV-1)- and human immunodeficiency virus type 1 (HIV-1)-infected cells. To evaluate the effect of substituting a 1-S-alkyl thioether for a 1-O-alkyl ether, we synthesized a series of PFA conjugates of 1-S-alkyl-sn-thioglycerols with varied 1-S-alkyl chain lengths. To establish structure-activity relationships we measured the in vitro antiviral activity of liposomal formulations of the drugs in cells infected with HCMV, HSV-1 or HIV-1. The optimum 1-S-alkyl chain length in the series was 16 to 18 carbon atoms. We compared the antiviral activity of 16- and 18-carbon alkyl thioglycerol versus alkylglycerol prodrugs and did not observe any significant differences in their antiviral activities. The series' most active member, 1-S octadecyl-sn-glycero-3-foscarnet (ODSG-PFA) was 56-, eight- and 45-fold more active than PFA in HCMV-, HSV-1- and HIV-1-infected cells in vitro. The oral absorption of PFA and 1-S-octadecyl-sn-thioglycero-3-PFA was compared in mice by measuring plasma levels of 14C after oral administration of radiolabelled compounds. The peak plasma level of 14C was sevenfold higher following administration of [14C]ODSG-PFA than following an equimolar dose of [14C]PFA. Area-under-the-curve was 23-fold greater for ODSG-PFA than for PFA. Like previously reported alkyloxyether-lipid PFA conjugates, alkylthioether conjugates provided enhanced antiviral activity and oral bioavailability. However, S-ether conjugates may be metabolized differently than O-ether conjugates. More detailed in vivo pharmacokinetic evaluation of the alkyl-thioether-PFA conjugates is required. PMID- 9875376 TI - Synthesis and antiviral evaluation of SATE-foscarnet prodrugs and new foscarnet AZT conjugates. AB - The synthesis of a range of di- and triester derivatives of phosphonoformate (PFA; foscarnet) as potential lipophilic, membrane-soluble prodrugs is described. In addition to normal alkyl esters in the carboxylate and phosphonate residues of PFA, the bioreversible S-(pivaloyl)thioethyl (t-butyl-SATE) group was introduced in an attempt to deliver PFA after bioactivation inside the cells. Furthermore, PFA-AZT conjugates were prepared in order to develop combinational drugs. The key synthetic step was in all cases the formation of the P-C bond to build up the different PFA esters. In contrast to the diester derivatives, the triesters of PFA showed high hydrolytic instability during chromatographic purification. The compounds were evaluated in vitro for their ability to inhibit viruses in several tissue culture systems. All PFA alkyl di- and triesters proved poorly active or inactive against human immunodeficiency virus (HIV) and inactive against hepatitis B virus. In contrast, the PFA-AZT conjugates exhibited significant anti HIV activity. However, this activity was nearly completely lost in thymidine kinase-deficient cells, suggesting a fast unselective chemical hydrolysis of the conjugates to yield the nucleoside analogue AZT in the cell culture medium. Furthermore, no synergistic effect of PFA and AZT was observed. PMID- 9875377 TI - Thiazolo[4,5-d]pyrimidines. Part II. Synthesis and anti-human cytomegalovirus activity in vitro of certain acyclonucleosides and acyclonucleotides derived from the guanine analogue 5-aminothiazolo[4,5-d]pyrimidine-2,7(3H,6H)-dione. AB - The synthesis and in vitro antiviral activity of certain hydroxyalkoxymethyl, hydroxyalkyl, hydroxyalkenyl and phosphonoalkenyl derivatives of the guanine congener 5-aminothiazolo[4,5-d]pyrimidine-2,7(3H,6H)-dione are reported. The compounds of this study were selected for their structural similarity to acyclonucleosides with known anti-herpesvirus activity. 5-Amino-3-[(Z)-4-hydroxy 2-buten-1-yl]thiazolo[4,5-d]pyrimidine-2, 7(3H,6H)- dione was the only member of the series to display significant in vitro activity against human cytomegalovirus (HCMV); however, this compound did not inhibit other herpesviruses, human immunodeficiency virus type 1 or murine cytomegalovirus. It was found to have a cytotoxicity profile similar to that of ganciclovir (DHPG). The antiviral effect was found to be sensitive to the initial viral input and the time of addition during the virus replication cycle. Significantly, the compound was found to have equal anti-HCMV activity, against standard virus strains, to DHPG, but also showed potent activity against DHPG-resistant virus strains, except for a strain mutated in the UL97 (phosphotransferase) gene. PMID- 9875378 TI - The hepatitis B virus M539V polymerase variation responsible for 3TC resistance also confers cross-resistance to other nucleoside analogues. AB - A variant of hepatitis B virus (HBV) containing a Met-to-Val substitution (M539V) in the YMDD motif of the polymerase nucleoside-binding domain exhibited resistance to the cytosine analogue lamivudine (3TC). To determine if the mutation responsible for the M539V polymerase variant affected the sensitivity of the virus to other nucleoside analogues, we constructed a tetracycline-responsive cell line, HepAD79. This cell line is stably transfected with a cDNA copy of the pregenomic RNA of an HBV genome containing an A-to-G mutation in the first position of the polymerase gene codon 539. This mutation results in a Met-to-Val substitution at amino acid 539 of the polymerase. When grown under the proper conditions, HepAD79 cells produced HBV RNA, contained HBV DNA associated with immature core particles and released core-associated HBV DNA into the medium. The M539V polymerase variant produced in this cell line was approximately 26-fold less sensitive to the antiviral effects of 3TC than wild-type virus. In addition, this variant demonstrated decreased sensitivity to the cytosine analogues FTC and ddC, as well as the thymidine analogue AZT. PMID- 9875379 TI - Design, synthesis and structure relationships of new N,N',N",N"'-tetrakis (omega amino alkyl) tetraazamacrocycles. AB - A number of N,N',N",N"'-tetrakis (omega-aminoalkyl) tetraazamacrocycles and related compounds were synthesized and evaluated for their inhibitory effects on human immunodeficiency virus type 1 (HIV-1) and duck hepatitis B virus (DHBV) replication. The activity of these compounds was found to be highly dependent upon structural features: (i) the length of the alkyl linker connecting the nitrogen atoms of the macrocyclic ring to the exocyclic nitrogen atoms of the terminal amino groups (five methylenes favoured antiviral activity); (ii) substitution of the terminal amino groups of the linker reduced antiviral activity; and (iii) the size of the tetraazamacrocyclic ring (14 or 15 atoms) did not markedly affect the antiviral activity. Some analogues were potent inhibitors of HIV-1 replication, with anti-HIV activity similar to that of biscyclam (JM 2763). In contrast, other analogues were found to be highly toxic in duck hepatocyte primary culture, the 2.2.15 cell line and to a lesser extent in MT-4 cells. Structural parameters, macrocyclic ring size and metal-chelating ability have been used to develop a structure-activity relationship model in order to aid the design of antiviral molecules derived from N,N',N",N"'-tetrakis (omega aminoalkyl) tetraazamacrocycles. PMID- 9875380 TI - Antiviral effects of 6-chloro-2',3'-dideoxyguanosine in rhesus monkeys acutely infected with simian immunodeficiency virus. AB - A lipophilic dideoxynucleoside analogue, 6-chloro-2',3'-dideoxyguanosine (6-Cl ddG), was expected to be effective against AIDS-related dementia. In this study, we tested the effect of 6-Cl-ddG on simian immunodeficiency virus (SIVmac239) replication in vitro and on acute infection of six rhesus monkeys (Macaca mulatta) with SIVmac239. This compound inhibited SIV-induced cytopathic effect in CEM x 174 cells and SIV replication in vitro with an ED50 value of 2.5 microM. A dose of 25 mg/kg 6-Cl-ddG was administered to three monkeys every 8 h for 10 days and an untreated group of three monkeys was injected with the solvent without drug. Although 6-Cl-ddG was not detected in the plasma, the metabolite ddG was maintained at a concentration of more than 3 microM for 8 h after administration. In the cerebrospinal fluid, the ddG concentration was 2 microM at 2 h after administration. SIV antigen (p27) and antibody appearance in the plasma were delayed for 5-8 days compared with the mock-treated group. The occurrence of lymphadenopathy in treated monkeys was delayed for 6 days compared with the mock treated group. Signs of 6-Cl-ddG toxicity were minimal after the treatment. The results of this study provide further evidence that 6-Cl-ddG may act as a potent anti-human immunodeficiency virus agent in vivo. PMID- 9875381 TI - Approaches to antiviral chemotherapy for acute respiratory infections. AB - The causative agents of acute respiratory infections (ARI) in infants and children are mostly thought to be viruses. Some ARI in adult patients may be caused by bacteria but most often the causes are virus infections. When ARI affect immunocompromised patients or the elderly the mortality rates are significantly higher than in immunocompetent individuals. Many types of viruses cause ARI. Among them, influenza viruses A and B and respiratory syncytial virus (RSV) are thought to be the most important because of the severity of illness after infection and their high communicability in the human population. Recently, several novel antiviral drugs against ARI have been developed and some are proceeding in clinical trials. This review covers current investigations into antiviral compounds targeted at several points in the virus life-cycle. This includes PM-523, which broadly inhibits ortho- and paramyxo-viruses, two neuraminidase inhibitors for influenza virus, neutralizing antibody to RSV and chimeric soluble ICAM-1-IgA molecules targeted against rhinoviruses. PMID- 9875382 TI - Synthesis and anti-HIV activity of some novel chain-extended phosphoramidate derivatives of d4T (stavudine): esterase hydrolysis as a rapid predictive test for antiviral potency. AB - Novel chain-extended nucleoside phosphoramidates of the anti-human immunodeficiency virus (HIV) drug d4T (stavudine) have been prepared as possible membrane-permeable prodrugs of the bio-active free 5'-monophosphates. Phosphorochloridate chemistry gave the target compounds in moderate to high yields, and all materials were fully characterized by spectroscopic and analytical methods. The compounds are related to the previously reported phenyl methoxyalaninyl derivative of d4T, which was shown to be a potent and selective inhibitor of HIV. In this study the amino acid nitrogen and ester moieties were separated by methylene spacers of between two and six carbon atoms. In vitro evaluation of these compounds indicated an almost complete lack of anti-HIV activity, the compounds being several orders of magnitude less potent than the corresponding alpha-amino acid derivatives. The reasons for the virtual lack of anti-HIV activity appear to involve poor enzyme-mediated hydrolysis. PMID- 9875383 TI - In vitro anti-human immunodeficiency virus and anti-hepatitis B virus activities and pharmacokinetic properties of heterodinucleoside phosphates containing AZT or ddC. AB - In vitro activities, against human immunodeficiency virus (HIV)- and hepatitis B virus (HBV)-infected cells, of four amphiphilic heterodinucleoside phosphates containing 3'-azido-2',3'-dideoxythymidine (AZT) or 2',3'-dideoxycytidine (ddC) as antiviral monomers were evaluated. The four compounds were N4-hexadecyl-2' deoxyribocytidylyl-(3'-->5')-3'-azido-2',3'-deoxyt hymidine (N4-hxddC-AZT), N4 palmitoyl-2'-deoxyribocytidylyl-(3'-->5')-3'-azido-2',3'-deoxyt hymidine (N4 pamdC-AZT), N4-hexadecyl-2'-deoxycytidylyl-(3'-->5')-2',3'-dideoxycytidine (N4 hxddC-ddC) and 2'-deoxythymidylyl-(3'-->5')-N4-palmitoyl-2',3'-dideoxycytidine (dT-N4-pamddC). All four dimers were active against HIV, dT-N4-pamddC being the most active and least toxic. dT-N4-pamddC also exhibited strong antiviral effects against a panel of eight AZT-resistant HIV strains. The ddC-containing heterodimers incorporated in liposomes additionally inhibited HBV replication by 50-80% in HepG2 2.2.15 cells. AZT and the AZT-containing dimers were ineffective. Differences in pharmacokinetic properties between the antiviral monomers and the heterodimers were evaluated using liposomal formulations of 3H-labelled AZT heterodimers as model compounds. The cellular distribution of AZT in H9 cells was predominantly cytoplasmic, whereas the amphiphilic dimers were distributed more evenly throughout the cytoplasm, nuclear membranes and microsomes. Blood levels of the heterodimers decreased at a rate two- to threefold slower than AZT and the areas-under-the-curves were five- to sevenfold higher for N4-pamdC-AZT and N4 hxddC-AZT, respectively. Compared to AZT, the peak levels of the dimers were three to four times higher in blood and five to six times higher in the liver. Analysis of blood samples showed that 34% of N4-pamdC-AZT was metabolized to AZT, whereas only 9% of N4-hxddC-AZT released AZT. Considering the antiviral potency and the favourable pharmacokinetic properties of the heterodimers, these compounds merit further exploration as antiviral drugs. PMID- 9875384 TI - 1,2,5-Benzothiadiazepine and pyrrolo[2,1-d]-[1,2,5]benzothiadiazepine derivatives with specific anti-human immunodeficiency virus type 1 activity. AB - We synthesized and tested as novel inhibitors of human immunodeficiency virus type 1 (HIV-1) bi- and tricyclic thiadiazine ring homologues of 7-chloro-2-ethyl 2H-1,2,4-benzothiadiazin-3-(4H)-one 1,1-dioxide, which is a compound endowed with anti-HIV-1 activity at low micromolar concentrations. Benzothiadiazepine derivatives were obtained by alkylation of 8-chloro-2,3-dihydro-3-methyl-1,2,5 benzothiadiazepin-4(5H)-one 1,1-dioxide, which was obtained by intramolecular cyclization of 2-(2-amino-5-chloro-benzenesulphonamido) propanoic acid. Pyrrolobenzothiadiazepines were synthesized from N-substituted 5-chloro-2-(1H pyrrol-1-yl)benzene-sulphonamides by treating with triphosgene. N6-substituted pyrrolo[2,1-d][1,2,5]benzothiazepin-7(6H)-one 5,5-dioxides were active, though not very potent. Both a chlorine atom and an unsaturated alkyl chain were found to be determinants of anti-HIV-1 activity. The highest potency was shown by a derivative with a TIBO-related 3,3-dimethylallyl chain. 2,3-Dihydro-1,2,5 benzothiadiazepin-4(5H)-one 1,1-dioxides were scarcely active in HIV-1-infected MT-4 cell assays; however, the introduction of the dimethylallyl chain into 7 chloro-1,2,5-benzothiadiazepine moiety led to a bicyclic derivative which was more potent and less cytotoxic than the tricyclic pyrrole-containing counterpart. PMID- 9875385 TI - Synthesis and biological evaluation of 5H-indolo [3,2-b][1,5]benzothiazepine derivatives, designed as conformationally constrained analogues of the human immunodeficiency virus type 1 reverse transcriptase inhibitor L-737,126. AB - In the presence of sodium hydride, reaction of aryl-disulphides with ethyl esters of indole-2-carboxylic acids furnished ethyl 3-arylthioindole-2-carboxylates, which were cyclized intramolecularly to afford 5H-indolo[3,2 b][1,5]benzothiazepin-6(7H)-ones or hydrolysed in alkaline medium to give 3 arylthioindole-2-carboxylic acids. These acids, also obtained by the action of aryldisulphides on indole-2-carboxylic acids, afforded tetracyclic 5H-indolo [3,2 b][1,5]benzothiazepin-6(7H)-ones upon treatment with EDCI-DMAP. Transformation of cyclic sulphides into the required sulphones was achieved by treatment with hydrogen peroxide or with m-chloroperbenzoic acid. The title derivatives are conformationally constrained analogues of the potent human immunodeficiency virus type 1 (HIV-1) reverse transcriptase inhibitor 3-benzene-sulphonyl-5-chloroindole 2-carboxamide (L-737, 126). Although the indolobenzothiazepine derivatives, as well as the indolyl aryl sulphones used for their synthesis, were endowed with anti-HIV-1 activities in the submicromolar and micromolar range, none of them proved more potent than L-737,126. PMID- 9875386 TI - Isolation and characterization of the fungal metabolite 3-O-methylviridicatin as an inhibitor of tumour necrosis factor alpha-induced human immunodeficiency virus replication. AB - The cytokine tumour necrosis factor alpha (TNF-alpha) has been shown to play a role in human immunodeficiency virus (HIV) replication by activating transcription of the provirus in both T cells and macrophages. Therefore, agents that block TNF-alpha-induced HIV expression could have therapeutic value in the treatment of AIDS. We have sought to identify antiviral agents that block TNF alpha induction of HIV LTR-directed transcription, using a cell-based, virus-free assay system in automated high-throughput screening. HeLa cells were transfected with an HIV LTR-luciferase reporter plasmid and a stable line was isolated in which TNF-alpha increased luciferase production by two- to threefold. This cell line was used to screen approximately 15,000 fungal extracts. An inhibitory activity specific for TNF-alpha-induced HIV LTR transcription was observed in culture OS-F67406. The active component was isolated and identified as a known metabolite, 3-O-methylviridicatin, by NMR and mass spectrometry. No biological activity has been associated with this compound previously. This compound blocks TNF-alpha activation of the HIV LTR in the HeLa-based system, with an IC50 of 5 microM, and inhibited virus production in the OM-10.1 cell line, a model of chronic infection responsive to induction by TNF-alpha, with an IC50 of 2.5 microM. PMID- 9875387 TI - In vitro evaluation and characterization of newly designed alkylamidophospholipid analogues as anti-human immunodeficiency virus type 1 agents. AB - Our laboratories first reported two novel classes of complex synthetic lipids, including alkylamidophosphocholines (PC lipid; CP-51) and alkylamidophosphate ester-linked lipid-AZT conjugates (lipid-AZT conjugates; CP-92), with selective and potent activity against human immunodeficiency virus type 1 (HIV-1). To extend these observations, we synthesized additional PC lipids and lipid-AZT conjugates (INK and INK-AZT conjugate) to evaluate their structure-activity relationships by testing for selectivity against infectious wild-type (wt) and drug-resistant HIV-1 replication, virus fusogenic activity and toxicity for mouse bone marrow cells. PC lipid compounds with medium chain lengths at positions 1 and 2 gave an improved selective index (SI). INK-3, with 12 and 8 carbons and INK 15, with 10 and 12 carbons were among the most selective when evaluated in CEM-SS cells. INK-14, a lipid-AZT conjugate where AZT replaced the choline in PC lipid INK-3, gave the highest SI of > 1250 against both infectious wt HIV-1 replication in CEM-SS cells and a clinical isolate in peripheral blood leukocytes. Notably, the PC lipid compounds INK-3 and INK-15, but not the lipid-AZT conjugate INK-14, were potent inhibitors of matched pairs of AZT-sensitive and AZT-resistant HIV-1 clinical isolates. INK-3 also inhibited replication of HIV-2 and TIBO-resistant HIV-1, and inhibited HIV-1-mediated fusogenic activity by 78, 41 and 9% in a dose dependent manner. The TC50 for mouse bone marrow cells was > 100 micrograms/ml for INK-3 compared to 9.15-14.17 micrograms/ml for CP-51 and 0.142-0.259 microgram/ml for AZT. These data suggest that optimum PC lipid compounds are significantly less toxic than AZT and have high potential as novel therapeutic agents for AIDS. PMID- 9875388 TI - Colorimetric assays for evaluation of the mode of action of human immunodeficiency virus type 1 non-nucleoside reverse transcriptase inhibitors. AB - Four non-nucleoside reverse transcriptase (RT) inhibitors, 9-CI-TIBO [(+)-S 4,5,6,7-tetrahydro-9- chloro-5-methyl-6-(3-methyl-2-butenyl)imidazo(4,5,1 jk)(1,4)- benzodiazepin-2(1H)-thione)], nevirapine (6,11-dihydro-11-cyclopropyl-4 methyl-dipyrido[2,3-b:2',3'-e]-[1,4]di azepin- 6-one), MSA-300 (N-[cis-2-(2 hydroxy-3-acetyl-6-methoxy-phenyl)-cyclopropyl]-N'- (5-chloropyrid-2-yl) thiourea) and delavirdine ?1-(5-methanesulphonamido-1H-indol-2-yl-carbonyl)-4-[3- (1-methylethylamino)pyridinyl]piperazine? were analysed for the mode of action of their inhibition of human immunodeficiency virus type 1 (HIV-1) RT in three different assays utilizing a 96-well microtitre plate format, with solid-phase conjugated poly(rA) as template. These were: (i) direct RT assay, for determination of IC50 values of RT inhibitors; (ii) RT template/primer binding inhibition (BIC) assay, for measuring the effect of various substances on the RT activity binding to template/primer; (iii) RT protein ELISA, for measuring RT protein binding to template/primer with a monoclonal antibody reactive against a peptide in the RNase H region. MSA-300 and delavirdine gave the lowest IC50 values, ranging from 0.17 microM to 0.24 microM for MSA-300 and from 0.12 microM to 0.38 microM for delavirdine, whereas higher IC50 values of approximately 20 microM were obtained for 9-CI-TIBO at all primer concentrations. None of the non nucleoside substances had inhibiting effects on the binding of template, primer, or template/primer to RT protein. Their inhibition of RT activity was not due to prevention of RT binding to template/primer. TIBO, nevirapine and delavirdine bound to RT reversibly, and they bound more tightly to RT template/primer ternary than to RT template binary complex. MSA-300 showed a comparatively high affinity for the enzyme. The utility of the three assays in relation to screening and analysis of RT inhibitory substances is discussed. PMID- 9875389 TI - 3-Hydroxyphthaloyl beta-lactoglobulin. III. Antiviral activity against herpesviruses. AB - The spread of sexually transmitted diseases, including human immunodeficiency virus type 1 (HIV-1) and herpesvirus infections, has continued unabated despite educational efforts spearheaded as a response to the HIV-1 epidemic. This suggests the need for prophylactic measures, including the application of topical antiviral agents. Chemical modification of bovine beta-lactoglobulin (beta-LG), the major protein of whey, by hydroxyphthalic anhydride (3HP) led to the generation of a potent HIV-1 inhibitor (designated 3HP-beta-LG) shown to also have activity against herpes simplex virus types 1 and 2 (HSV-1, HSV-2). This report provides more detailed results concerning the anti-herpesvirus activity of 3HP-beta-LG, indicating that this compound: (i) inhibited infection by human cytomegalovirus (HCMV), which is known to be sexually transmitted; (ii) inactivated the infectivity of both HSV-1 and HSV-2; (iii) inhibited cell-to-cell transmission of HSV-1 and HSV-2; and (iv) bound to HSV-1, HSV-2 and HCMV virus particles and partially inhibited the binding of anti-glycoprotein E (gE) and anti-gC monoclonal antibodies to HSV-1 and HSV-2. The binding of 3HP-beta-LG to the herpesviruses under study was inhibited by aggregated human IgG, suggesting that the respective viral Fc receptor is one of the target sites for 3HP-beta-LG. In agreement with results on inhibition of HIV-1 infection, 3HP-beta-LG appears to be the acid anhydride-modified protein of choice as an antiviral agent against herpesviruses. PMID- 9875390 TI - Molecular targets for human papillomaviruses: prospects for antiviral therapy. AB - A substantial medical need exists for the development of antiviral medicines for the treatment of diseases associated with infection by human papillomaviruses (HPVs). HPVs are associated with various benign and malignant lesions including benign genital condyloma, common skin warts, laryngeal papillomas and anogenital cancer. Since treatment options are limited and typically not very satisfactory, the development of safe and effective antiviral drugs for HPV could have substantial clinical impact. In the last few years, exciting advances have been made in our understanding of papillomavirus replication and the effects that the virus has on growth of the host cell. Although still somewhat rudimentary, techniques have been developed for limited virion production in vitro offering the promise of more rapid advances in the dissection and understanding of the virus life cycle. Of the 8-10 HPV gene products that are made during infection, only one encodes enzymatic activities, the E1 helicase. Successful antiviral therapies have traditionally targeted viral enzymes such as polymerases, kinases and proteases. In contrast, macromolecular interactions which mediate the functions of E6, E7 and E2 are thought to be more difficult targets for small molecule therapy. PMID- 9875391 TI - Potent beta-lactam inhibitors of human cytomegalovirus protease. AB - A series of novel monobactam inhibitors of human cytomegalovirus (HCMV) protease has been described that possess a heterocyclic thiomethyl side chain at C-4. Changes to the heterocycle did not significantly change the inhibitory activity of these compounds in an enzymatic assay, although improvements in solubility and cell culture activity were noted. A number of permutations between C-4 substitutions and N-1 derivatives led to the identification of several beta lactams with antiviral activity in a plaque reduction assay. N-methyl thiotetrazole-containing compounds were found to be the most potent inhibitors in the enzymatic assay. PMID- 9875392 TI - Chemistry and anti-herpes simplex virus type 1 evaluation of cycloSal-nucleotides of acyclic nucleoside analogues. AB - The synthesis of different cycloSal-phosphotriesters of the acyclic nucleoside analogues acyclovir (ACV), penciclovir (PCV) and T-penciclovir (T-PCV) as potential new lipophilic, membrane-soluble pronucleotides is described. The introduction of the cycloSal moiety was achieved by using reactive cyclic chlorophosphane reagents. In addition to the cycloSal-PCV monophosphate (MP) phosphotriesters, a second derivative bearing an acetyl group at the second primary alcohol function was prepared. In hydrolysis studies the cycloSal-ACVMPs showed the expected range of hydrolytic stability dependent on the substituent in the masking group (8-17 h). In contrast, the cycloSal-PCVMP derivatives exhibited a 11- to 15-fold increase in hydrolytic lability as compared to the corresponding cycloSal-ACVMP derivatives. We demonstrated that the free primary alcohol group is responsible for this rate acceleration because cycloSal-OAc-PCVMP, in which the hydroxyl group was blocked by acetylation, did not show the aforementioned acceleration. Unexpectedly, the hydrolysis product was not PCVMP but according to NMR and mass spectrometry it was cycloPCVMP (cPCVMP). The title compounds were evaluated in vitro for their ability to inhibit herpes simplex virus type 1 (HSV 1) and thymidine kinase-negative (TK-) HSV-1 replication in Vero cells. The cycloSal-ACVMP compounds exhibited high antiviral activity in HSV-1-infected cells. More importantly, one derivative retained all activity from the wild-type virus strain in HSV-1/TK(-)-infected Vero cells. The PCV derivatives were markedly less active. The reason for the failure of the cycloSal-PCVMPs seems to be due to the formation of cPCVMP instead of the desired PCVMP. PMID- 9875393 TI - Broad-spectrum antiviral activity and mechanism of antiviral action of the fluoroquinolone derivative K-12. AB - The fluoroquinolone derivatives have been shown to inhibit human immunodeficiency virus (HIV) replication at the transcriptional level. We confirmed the anti-HIV activity of the most potent congener, 8-difluoromethoxy-1-ethyl-6-fluoro-1,4 dihydro-7-[4-(2- methoxyphenyl)-1-piperazinyl]-4-quinolone-3-carboxylic acid (K 12), in both acutely and chronically infected cells. K-12 was active against different strains of HIV-1 (including AZT- and ritonavir-resistant HIV-1 strains), HIV-2 and simian immunodeficiency virus, in MT-4, CEM, C8166 and peripheral blood mononuclear cells. In all of these antiviral assay systems, K-12 showed a similar activity (EC50 0.2-0.6 microM). K-12 inhibited Moloney murine sarcoma virus-induced transformation of C3H/3T3 cells with an EC50 of 6.9 microM. Also, K-12 proved inhibitory to herpesvirus saimiri, human cytomegalovirus, varicella-zoster virus and herpes simplex virus types 1 and 2 (in order of decreasing sensitivity), but was not inhibitory (at subtoxic concentrations) to human herpesvirus type 8 (as evaluated in BCBL-1 cells), vaccinia virus, Sindbis virus, vesicular stomatitis virus, respiratory syncytial virus, Coxsackie virus, Punta Toro virus, parainfluenza virus or reovirus. Time-of-addition experiments and quantitative transactivation bioassays indicated that K-12 inhibits the Tat mediated transactivation process in HIV-infected cells. PMID- 9875394 TI - An approach towards the synthesis of potential metal-chelating TSAO-T derivatives as bidentate inhibitors of human immunodeficiency virus type 1 reverse transcriptase. AB - Novel derivatives of the potent human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) inhibitor TSAO-T have been designed, synthesized and tested for their in vitro antiretroviral activity against HIV. These TSAO-T derivatives have been designed as potential bidentate inhibitors of HIV-1 RT, which combine in their structure the functionality of a non-nucleoside RT inhibitor (TSAO-T) and a bivalent ion-chelating moiety (a beta-diketone moiety) linked through an appropriate spacer to the N-3 of thymine of TSAO-T. Some of the new compounds have an anti-HIV-1 activity comparable to that of the parent compound TSAO-T, but display a markedly increased antiviral selectivity. There was a clear relationship between antiviral activity and the length of the spacer group that links the TSAO molecule with the chelating moiety. A shorter spacer invariably resulted in increased antiviral potency. None of the TSAO-T derivatives were endowed with anti-HIV-2 activity. PMID- 9875395 TI - 3-(5-Dimethylamino-1-naphthalenesulphonyl)-2-(3-pyridyl)thiazolidine (YHI-1) selectively inhibits human immunodeficiency virus type 1. AB - 3-(5-Dimethylamino-1-naphthalenesulphonyl)-2-(3-pyridyl)thiazolidi ne (YHI-1), a synthetic analogue of D-cysteinolic acid isolated from sardines (Sardinops melanostictus), was found to be a specific inhibitor of human immunodeficiency virus type 1 (HIV-1) replication in various cell cultures. YHI-1 inhibited HIV 1IIIB replication with a 50% effective concentration (EC50) of 3.35, 10.23 and 4.61 microM in MT-4 cells, peripheral blood mononuclear cells and MAGI-CCR5 cells, respectively. However, no antiviral activity was observed with non nucleoside reverse transcriptase inhibitor (NNRTI)-resistant HIV-1 strains, such as nevirapine-resistant HIV-1HE/NEV and MKC-442-resistant HIV-1IIIB-R, or with HIV-2ROD or SIVMAC. YHI-1 failed to inhibit reverse transcriptase (RT) activity in vitro with different template-primer systems. Time-of-addition experiments, the failure to inhibit NNRTI-resistant strains and the failure to show in vitro activity against RT suggest that a metabolite of YHI-1 inside the cell acts like an NNRTI. Thus, YHI-1 seems to belong to a new class of HIV-1 inhibitor and is a good candidate for further development. PMID- 9875396 TI - Synthesis, structure and in vitro anti-human immunodeficiency virus activity of novel 3-methyl-1H,3H-thiazolo[3,4-a]benzimidazoles. AB - A series of novel 1-aryl-3-methyl-1H,3H-thiazolo[3,4-a]benzimidazoles, TBZ analogues, were synthesized and investigated as anti-human immunodeficiency virus (HIV) agents in order to study the effects of structural modifications on antiviral activity and cytotoxicity. They were proved to inhibit significantly HIV-1 replication in vitro without showing inhibitory activity on HIV-2 or simian immunodeficiency virus. Their potency was influenced by the presence of suitable substituents in the phenyl ring at C-1 as well as by their stereochemical characteristics. In fact, the most active compound of the series was the trans-1 (2,6-difluorophenyl)-3-methyl-1H,3H-thiazolo[3,4- a]benzimidazole, in which the butterfly-like conformation is stabilized by two intramolecular hydrogen bonds between the fluorine atoms and H-1 and H-3. This was made possible by the trans arrangement of C-1 and C-3 substituents, as shown by X-ray and NMR analysis. PMID- 9875397 TI - Aryl phosphate derivatives of 3'-deoxythymidine are not potent anti-human immunodeficiency virus agents. AB - Aryl phosphate derivatives of 3'-deoxythymidine (3dT), albeit more active than 3dT in thymidine kinase (TK)-deficient cells, are not potent anti-human immunodeficiency virus (HIV) agents and are capable of inhibiting HIV replication only at micromolar concentrations. PMID- 9875398 TI - D4T-5'-[p-bromophenyl methoxyalaninyl phosphate] as a potent and non-toxic anti human immunodeficiency virus agent. AB - Three aryl phosphate derivatives of 2',3'-didehydro-2',3'-dideoxythymidine (d4T) were tested for their anti-human immunodeficiency virus (HIV) activity in peripheral blood mononuclear cells (PBMC) and thymidine kinase (TK)-deficient CEM T cells. Compared to the parent compound d4T, the lead compound d4T-5'-[p bromophenyl methoxyalaninylphosphate] with a para-bromo substituent in the aryl moiety was 12.6-fold more potent in inhibiting p24 production (IC50 values: 44 nM versus 556 nM) and 41.3-fold more potent in inhibiting the reverse transcriptase (RT) activity (IC50 values: 57 nM versus 2355 nM) in HIV-infected TK-deficient CEM cells. None of the compounds exhibited any detectable cytotoxicity to PBMC or CEM cells at concentrations as high as 10,000 nM. To our knowledge, this is the first demonstration that the potency as well as selectivity index of the d4T aryl phosphate derivatives in TK-deficient cells can be substantially enhanced by introducing a single para-bromo substituent in the phenyl moiety. PMID- 9875399 TI - Anti-human immunodeficiency virus drug combination strategies. AB - It is now generally accepted that mono- and bitherapy for human immunodeficiency virus type 1 (HIV-1) infection are only transiently efficient mainly due to virus drug resistance. To obtain a sustained benefit from antiviral therapy, current guidelines recommend at least triple-drug combinations, or the so-called highly active antiretroviral therapy (HAART). In some patients, HAART can be problematic, either because it is difficult to remain compliant or because previous suboptimum therapies have limited the choice of drugs. For compliant drug-naive patients, HAART should be able to offer long-term virus suppression, when changing from first- to second- to third-line HAART at drug failure. Long term treatment might ultimately result in multi-drug resistant virus leaving few options for salvage therapy. HIV drug resistance testing to guide this salvage therapy and the development of new drugs to allow new options will therefore remain priorities in anti-HIV drug research. PMID- 9875400 TI - Imidazo[1,5-b]pyridazine-d4T conjugates: synthesis and anti-human immunodeficiency virus evaluation. AB - In an attempt to combine the human immunodeficiency virus type 1 (HIV-1) inhibitory capacity of 2',3'-dideoxy-2',3'-didehydronucleoside analogues [nucleoside reverse transcriptase (RT) inhibitors; NRTI] and non-nucleoside RT inhibitors (NNRTI), we have designed, synthesized and evaluated for their anti HIV activity several heterodimers of the general formula [d4T]-NH-(CH2)n-NH [imidazo[1,5-b]pyridazine]. The synthesis of these heterodimers was conducted in three parts. The first part focused on the synthesis of the NRTI. The second part was devoted to the NNRTI and the NNRTI linked to appropriate spacers: [NNRTI]-NH (CH2)n-NH2. In the third part, the condensation between the NRTI and the [NNRTI] NH-(CH2)n-NH2 was performed. The in vitro inhibitory activities against HIV-1 of the [d4T]-NH-(CH2)n-NH-[imidazo[1,5-b]pyridazine] heterodimers were found to be comparable to that of d4T (stavudine) in HIV-infected cells. Moreover, the heterodimers were endowed with anti-HIV-2 activity and with anti-nevirapine resistant HIV-1 activity. None of the heterodimers proved markedly cytotoxic to CEM-SS or MT-4 cells. There was not a clear trend toward antiviral potency on lengthening the methylene spacer in the [d4T]-NH-(CH2)n-NH-[imidazo[1,5 b]pyridazine] heterodimers. PMID- 9875401 TI - Targeting RNase L to human immunodeficiency virus RNA with 2-5A-antisense. AB - In an attempt to develop a lead for the application of 2-5A-antisense to the targeted destruction of human immunodeficiency virus (HIV) RNA, specific target sequences within the HIV mRNAs were identified by analysis of the theoretical secondary structure. 2-5A-antisense chimeras were chosen against a total of 11 different sequences: three in the gag mRNA, three in the rev mRNA and five in the tat mRNA. 2-5A-antisense chimera synthesis was accomplished using solid-phase phosphoramidite chemistry. These chimeras were evaluated for their activity in a cell-free assay system using purified recombinant human RNase L to effect cleavage of 32P-labelled RNA transcripts of plasmids derived from HIV NL4-3. This screening revealed that of the three 2-5A-antisense chimeras targeted against gag mRNA, only one had significant HIV RNA cleavage activity, approximately 10-fold reduced compared to the parent 2-5A tetramer and comparable to that reported for the prototypical 2-5A-anti-PKR chimera, targeted against PKR mRNA. The cleavage activity of this chimera was specific, since a scrambled antisense domain chimera and a chimera without the key 5'-monophosphate moiety were both inactive. The 10 other 2-5A-antisense chimeras against tat and rev had significantly less activity. These results imply that HIV gag RNA, like PKR RNA and a model HIV tat oligoA-vif RNA, can be cleaved using the 2-5A-antisense approach. The results further imply that not all regions of a potential RNA target are accessible to the 2-5A-antisense approach. PMID- 9875402 TI - Synthesis and evaluation of some masked phosphate esters of the anti-herpesvirus drug 882C (netivudine) as potential antiviral agents. AB - A number of symmetric and asymmetric 5'-phosphate esters of the potent anti varicella-zoster virus (VZV) agent 1-(beta-D-arabinofuranosyl)-5-prop-1 ynyluracil (882C; netivudine) were prepared as potential lipophilic, membrane soluble prodrugs of the bio-active phosphate forms. The compounds were prepared by the base-catalysed coupling of various phosphorochloridates with the free nucleoside analogue. Compounds were fully characterized by a range of spectroscopic and analytical methods and were studied for their inhibition of several viruses in tissue culture. All of the phosphate esters were inactive against human cytomegalovirus, herpes simplex virus type 2, VZV, human immunodeficiency virus type 1 and influenza A virus (EC50 > 100 microM) except the 5'-(4-nitrophenyl phenyl) phosphate, which inhibited influenza A virus. The relative rate of esterase-mediated hydrolysis of one of the lead target structures was measured in order to rationalize the poor antiviral action, and data were collected on possible metabolites in support of this analysis. Cell specific esterases are implicated as key determinants of the antiviral potency of prodrugs of this type. PMID- 9875403 TI - Treatment of visna virus infection in lambs with the acyclic nucleoside phosphonate analogue 9-(2-phosphonylmethoxyethyl)adenine (PMEA). AB - Nucleoside and nucleotide analogues, which are inhibitors of human immunodeficiency virus reverse transcriptase, are highly active inhibitors of visna virus replication in cell cultures. One such analogue, the acyclic nucleoside phosphonate PMEA, has also been found to have a prophylactic effect on visna virus infection in lambs. In the present study, lambs were injected subcutaneously with 10 mg/kg PMEA three times a week starting 4 weeks after inoculation with visna virus, when brain infection had been established. After 3 weeks of treatment there was a reduction in the amount of virus isolated from blood cells of PMEA-treated lambs compared to controls and during the remaining 7 months of drug treatment there was significantly less virus isolated from the blood cells of treated lambs than of controls. Antibody response against visna virus was also slower in the treated than in the untreated control group. On the other hand, there was no difference in the amount of virus isolated from various organs of the two groups and the severity of CNS lesions in sheep treated with PMEA for 8 months was comparable to that found in untreated controls, even though PMEA reached concentrations in the cerebrospinal fluid which were well in excess of the EC50 value of the drug for visna virus. PMID- 9875404 TI - Specific inhibition of influenza virus RNA polymerase and nucleoprotein gene expression by liposomally encapsulated antisense phosphorothioate oligonucleotides in MDCK cells. AB - We have demonstrated that antisense phosphorothioate oligonucleotides (S-ODNs) inhibit influenza A virus replication in MDCK cells. Liposomally encapsulated and free antisense S-ODNs with four target sites (PB1, PB2, PA and NP genes) were tested for their abilities to inhibit virus-induced cytopathogenic effects in a MTT assay using MDCK cells. The liposomally encapsulated S-ODN complementary to the site around the PB2 AUG initiation codon showed highly inhibitory effects. In contrast, the inhibitory effect of the liposomally encapsulated S-ODN targeted to PB1 was considerably decreased in comparison with that directed to the PB2 target site. The liposomally encapsulated antisense S-ODNs exhibited higher inhibitory activities than the free oligonucleotides, and showed sequence-specific inhibition, whereas free antisense S-ODNs were observed to inhibit viral adsorption to MDCK cells. Liposomal preparations of oligonucleotides facilitated their release from endocytic vesicles, and thus cytoplasmic and nuclear localization was observed. The activities of the antisense S-ODNs were effectively enhanced by using the liposomal carrier. Interestingly, the liposomally encapsulated FITC-S-ODN-PB2-as accumulated in the nuclear region of MDCK cells. However, weak fluorescence was observed within the endosomes and the cytoplasm of MDCK cells treated with the free antisense S-ODNs. The cationic lipid particles may thus be a potentially useful delivery vehicle for oligonucleotide-based therapeutics and transgenes, appropriate for use in vitro or in vivo. PMID- 9875405 TI - Inhibitory effects of podophyllotoxin derivatives on herpes simplex virus replication. AB - Podophyllotoxin and its derivatives were examined for inhibitory effects on the replication of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), including acyclovir-resistant virus and clinical isolates. Deoxypodophyllotoxin (RD4-6266) proved to be a highly potent and selective inhibitor of all HSV strains in MRC-5 cells. EC50 values of RD4-6283 (in which the methylenedioxy ring A is modified) for HSV-1 and -2 were inferior to those of deoxypodophyllotoxin. However, podorhizol (RD4-6277) and 5'-methoxy-podorhizol (RD4-6276), in which ring C is absent, did not inhibit HSV replication. Moreover, RD4-6266 also inhibited the production of infectious virus particles of HSV-1 KOS strain and HSV-2 G strain. In contrast, none of the podophyllotoxin derivatives were found to have an antiviral effect against influenza A virus, respiratory syncytial virus or human cytomegalovirus in doses not toxic to the cells. PMID- 9875406 TI - Inhibition of human cytomegalovirus proteinase by salcomine derivatives. AB - Salcomine, N,N'-bis(salicylidene)ethylene diamino-cobalt (II), and its derivatives were evaluated for their ability to inhibit selectively human cytomegalovirus (HCMV) proteinase activity. The 50% inhibitory concentration (IC50) of salcomine was 1.4 microM for HCMV proteinase, but > 200 microM for three other serine proteinases (trypsin, > 250 microM; chymotrypsin, 206 microM; and elastase, > 250 microM). Two salcomine derivatives also inhibited HCMV proteinase with IC50 values under 2 microM. Studies of the structure-activity relationship of salcomine-related compounds showed that the phenyl moiety and the spacer moiety (distance between the two amines) were instrumental in the inhibition of HCMV proteinase. Moreover, salcomine inhibited the growth of laboratory strain AD169 and three clinical isolates at a 50% effective concentration (EC50) range of 1.92-2.89 microM. These results show that salcomine derivatives are potent and selective inhibitors of HCMV proteinase and HCMV replication in cell culture. Salcomine derivatives appear to be worth pursuing as candidate drugs for the chemotherapy of HCMV infection. PMID- 9875407 TI - Inhibitory effects of novel nucleoside and nucleotide analogues on Epstein-Barr virus replication. AB - The anti-Epstein-Barr virus (EBV) activity of different classes of compounds was assessed by means of an EBV DNA hybridization assay using a digoxigenin-labelled probe specific for the BamHI W fragment of the EBV genome, as well as by measuring viral capsid antigen (VCA) expression after a 7 day incubation period of P3HR-1 producer cells with the test substances. Acyclovir, ganciclovir, cidofovir and zidovudine were included as reference compounds. Several compounds proved to be potent and selective inhibitors of EBV DNA synthesis and VCA expression. Of the new compounds that were evaluated for their anti-EBV activity, the highest efficacy (lowest EC50) and highest selectivity index (SI) were shown by the purine nucleoside analogue 2-amino-7-[(1,3-dihydroxy-2 propoxy)methyl]purine (S2242) (EC50 0.6 ng/ml; SI 600), the acyclic nucleoside phosphonate analogues 9-(2-phosphono -methoxyethyl)-6-dimethylaminopurine (EC50 1.1 micrograms/ml; SI 91), 9-(2-phosphonomethoxyethyl)-2- amino-6 benzhydrylaminopurine (EC50 1.3 micrograms/ml; SI 29), 7-(2 phosphonomethoxyethyl)-6-dimethyl-aminopurine (EC50 0.8 microgram/ml; SI 56), 9 (R)-(2-phosphonomethoxypropyl)-6-(2-dimethylaminoethyl)-aminopur ine (EC50 0.5 microgram/ml; SI 42), the 2',3'-dideoxythymidine derivative 3'-oximino-2',3' dideoxythymidine (EC50 1.5 micrograms/ml; SI 65), and 1-(2,3- dideoxy-3-N hydroxyamino-beta-D-threo-pentafuranyl)pentafuranos yl)thymine (EC50 4.1 micrograms/ml; SI > 24). PMID- 9875408 TI - Strategies for antiviral drug discovery. AB - The need for antiviral drugs is growing rapidly as more viral diseases are recognized. The methods used to discover antiviral drugs have evolved considerably over the past 40 years and the overall process of discovery can be broken down into subprocesses which include lead generation, lead optimization and lead development. Various methods are now employed to ensure these processes are carried out efficiently. For lead generation, screening methodologies have developed to the extent where hundreds of thousands of compounds can be screened against a particular target. An alternative approach is to use the structures of enzyme substrates as a starting point for drug discovery. Much use is now made of X-ray crystallographic data of target-inhibitor complexes for the optimization of lead structures, and methods for preparing libraries of compounds to assist both generation and optimization of leads are well-developed. The methods used to predict and improve the pharmacokinetic properties of compounds are also changing rapidly. Finally, novel approaches to antiviral therapy using oligonucleotide based compounds or by modulating the host immune response are also being explored. This review discusses these approaches, provides examples of where their application has been successful and sets them against a historical background. PMID- 9875409 TI - Potent and selective inhibitors of human immunodeficiency virus protease structurally related to L-694,746. AB - A series of human immunodeficiency virus (HIV) protease inhibitors, which are analogues of N-[2(R)-hydroxy-1(S)- indanyl]-5(S)-[(tert-butyloxycarbonyl)amino] 4(S)-hydroxy-6-phenyl-2-(R) - [[4-(carboxymethoxy)phenyl]methyl]hexanamide (L 694,746), a metabolite of the anti-HIV agent L-689,502, were synthesized. In these compounds, the acetic group linked to the para position of the P1' phenyl in the reference inhibitor was replaced either by the bioisosteric phosphonomethoxy group and its diisopropyl/dibenzyl derivatives, or the 1H tetrazol-5-yl-methoxy group and its 1-benzyl derivative. In enzyme assays, phosphonomethoxy and tetrazolmethoxy analogues proved to be potent inhibitors of the HIV-1 protease, with IC50 values as low as 0.04 nM. When tested for anti-HIV 1 activity in cell-based assays, most of the new derivatives proved active, with benzyl derivatives being more active than their highly polar, unsubstituted counterparts. The dibenzylphosphonomethoxy analogue was the most active compound, with an EC50 value of 10 nM and a selectivity index of 20,000. When compounds were examined for their capability to reduce p24 levels in both acutely and chronically infected MT-4 and H9/IIIB cells, all of them were found to be active at concentrations close to those capable of preventing HIV-1-induced cytopathic effect. PMID- 9875410 TI - Synthesis, in vitro anti-human immunodeficiency virus structure-activity relationships and biological stability of 5'-O-myristoyl analogue derivatives of 3'-azido-2',3'-dideoxythymidine (AZT) as potential prodrugs. AB - 5'-O-Myristoyl analogue derivatives of 3'-azido-2',3'-dideoxythymidine (AZT), designed as potential double-barrelled prodrugs to AZT and the myristic acid analogues, were synthesized. Their ability to protect CEM cells against human immunodeficiency virus (HIV)-induced cytopathogenicity was determined and structure-activity paradigms were developed. 3'-Azido-2',3'-dideoxy-5'-O-(4 oxatetradecanoyl)thymidine (EC50 = 1.4 nM) and 3'-azido-2',3'-deoxy-5'-O-(12 bromododecanoyl)thymidine (EC50 = 3.2 nM) were the most effective anti-HIV-1 agents, relative to AZT (EC50 = 10 nM). These myristoyl analogue derivatives were more lipophilic (calculated log P = 4.5-8.1 range) than the parent compound AZT (log P = 0.06), and a linear correlation between their log P and HPLC log retention times was observed. The ester cleavage half-lives (t1/2) for esters upon in vitro incubation with porcine liver esterase, rat plasma or rat brain homogenate was dependent on the steric bulk, and electronegative inductive effect of the alpha-substituent (H, Br, F), of the 5'-O-myristoyl analogue moiety. 3' Azido-2',3'-dideoxy-5'-O-(11-(4-iodophenoxy) undecanoyl)-thymidine exhibited t1/2 values of 80.4, 3.7 and 150.0 min upon incubation with porcine liver esterase, rat plasma and rat brain homogenate, respectively. PMID- 9875411 TI - Allosteric inhibitors against HIV-1 reverse transcriptase: design and synthesis of MKC-442 analogues having an omega-functionalized acyclic structure. AB - Based on X-ray crystallographic analysis of MKC-442/human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) complex, analogues in which the N1 substituent is replaced with omega-functionalized alkyl groups were designed to improve the affinity for the enzyme. Synthesis of these compounds was carried out starting from MKC-442 by a sequence of reactions (N3-protection, removal of N1 ethoxymethyl group, alkylation, and N3-deprotection). The compounds were evaluated for anti-HIV activity. Structure-activity relationships are discussed in terms of the possible interaction with the enzyme. PMID- 9875412 TI - Novel 3'-spiro nucleoside analogues of TSAO-T. Part II. A comparative study based on NMR conformational analysis in solution and theoretical calculations. AB - The structures of two novel 3'-spiro nucleosides analogues of the potent human immunodeficiency virus type 1 (HIV-1) reverse trancriptase (RT) inhibitor TSAO m3T, in solution, as derived from NMR spectroscopy are described. In these TSAO analogues the spiro amino oxathioledioxide moiety has been replaced by spiro amino oxazolone or spiro amino oxathiazoledioxide moieties. A comparative study based on theoretical calculations of the hydrophobicity, the solvation free energies and molecular electrostatic potentials (MEP) of the three compounds is also described. No significant conformational differences were detected in solution between TSAO-m3T and its analogues that might account for the differences observed in their inhibitory activity against HIV-1 RT. The calculated hydrophobicity (log P) values, dipole moments and the electrostatic contributions to the solvation free energies of the three spiro ring systems were also similar. However, the differences found in the calculated MEPs of the spiro systems between TSAO-m3T and its analogues suggest that the different electrostatic surroundings of the 4"-amino group of the spiro moiety in the analogues may be responsible for a detrimental electrostatic interaction of the spiro rings with the Glu-B138 of RT. PMID- 9875413 TI - (Z)- and (E)-2-(hydroxymethylcyclopropylidene)-methylpurines and pyrimidines as antiviral agents. AB - Several Z- and E-methylenecyclopropane nucleoside analogues were synthesized and tested for antiviral activity in vitro against human and murine cytomegalovirus (HCMV, MCMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV), hepatitis B virus (HBV), herpes simplex virus types 1 and 2 (HSV-1, HSV-2), human herpesvirus 6 (HHV-6) and human immunodeficiency virus type 1 (HIV-1). The Z-2-amino-6 cyclopropylaminopurine analogue was the most effective agent against HCMV (EC50 or EC90 0.4-2 microM) followed by syncytol and the Z-2,6-diaminopurine analogues (EC50 or EC90 3.4-29 and 11-24 microM, respectively). The latter compound was also a strong inhibitor of MCMV (EC50 0.6 microM). Syncytol was the most potent against EBV (EC50 < 0.41 and 2.5 microM) followed by the Z-2,6-diaminopurine (EC50 1.5 and 6.9 microM) and the Z-2-amino-6-cyclopropyl-aminopurine derivative (EC50 11.8 microM). Syncytol was also most effective against VZV (EC50 3.6 microM). Activity against HSV-1, HSV-2 and HHV-6 was generally lower; synthymol had an EC50 of 2 microM against HSV-1 (ELISA) and 1.3 microM against EBV in Daudi cells but was inactive in other assays. The 2-amino-6-cyclopropylamino analogue displayed EC50 values between 215 and > 74 microM in HSV-1 and HSV-2 assays. 2 Amino-6-cyclopropylaminopurine and 2,6-diaminopurine derivatives were effective against HBV (EC50 2 and 10 microM, respectively), whereas none of the analogues inhibited HIV-1 at a higher virus load. Syncytol and the E isomer were equipotent against EBV in Daudi cells but the E isomer was much less effective in DNA hybridization assays. The E-2,6-diaminopurine analogue and E isomer of synthymol were devoid of antiviral activity. PMID- 9875414 TI - 3-Hydroxyphthaloyl beta-lactoglobulin. IV Antiviral activity in the mouse model of genital herpesvirus infection. AB - The spread of sexually transmitted infections caused by herpes simplex virus type 2 (HSV-2) has continued unabated despite educational efforts generated in response to the human immunodeficiency virus (HIV) epidemic. Given the absence of effective vaccines, this indicates the need to develop prophylactic measures such as topical antiviral agents. Chemical modification of bovine beta-lactoglobulin (beta-LG), the major protein of whey, by hydroxyphthalic anhydride (3HP) led to the generation of a potent HIV-1 inhibitor designated 3HP-beta-LG. This agent was shown to also have antiviral activity against HSV-2 and HSV-1 in vitro. Recent studies indicate that 3HP-beta-LG binds to HSV-1 virions, which, at least in part, involves the viral glycoprotein gE. Here we show that 3HP-beta-LG inhibits HSV-2 infection in the mouse model of genital HSV-2 infection. Simultaneous exposure to HSV-2 and 3HP-beta-LG caused a significant decrease in the proportion of infected animals (27% virus shedding, 5% lesion development and 0% fatality for 3HP-beta-LG as compared to 80% shedding, 60% lesion development and 53% fatality in mice treated with PBS). The proportion of animals with HSV-2 infection after treatment with beta-LG was similar to that in the PBS-treated group. Pretreatment with 3HP-beta-LG formulated in a gel, which prolongs the presence of the agent in the vagina, also significantly reduced the proportion of HSV-2-infected mice (5% virus shedding, 5% lesion development and 0% fatality for 3HP-beta-LG as compared to 70% shedding, 60% lesion development and 40% fatality in vehicle-treated mice). These differences were significant (P < or = 0.0005, 0.002 and 0.008 for shedding, lesion development and fatality, respectively). Virus titres in the minority of mice that developed infection were similar to those in untreated mice. HSV-2 infection was not inhibited by treatment of an ongoing infection, indicating that 3HP-beta-LG interferes with the initial infection. These data suggest that 3HP-beta-LG may be an efficacious agent for preventing vaginal transmission of genital herpesvirus infections. PMID- 9875415 TI - Current status of anti-HBV chemotherapy. AB - In the past decade, significant progress has been achieved in the battle against hepatitis B virus. In addition to the immunomodulating agents such as interferon alpha and thymosin, many novel antiviral agents have been discovered, among which nucleoside analogues are the mainstay. New-generation compounds such as 3TC and famciclovir have shown promise in the treatment of patients chronically infected by this virus, and are on the line for approval. However, viral rebound after cessation of therapy still remains a major problem. Additionally, the reports on the drug resistance to these antiviral agents suggest that combination therapy will be the eventual strategy (Bartholomew et al., 1997; Tipples et al., 1996). Therefore, developments of safe and effective antiviral agents which do not cross resist with currently available antiviral drugs are still much needed. PMID- 9875416 TI - Structure-activity relationship of fluoroquinolone in Escherichia coli. AB - Structure-activity relationship of 20 fluoroquinolones was studied using the susceptible and 4 resistant Escherichia coli which were developed against 4 fluoroquinolones [ciprofloxacin (1), KR-10755 (6), norfloxacin (2), and ofloxacin (3)] in our laboratory. The C-7 and C-8 substituents of fluoroquinolone were important in various functions such as the inhibitory activity on DNA gyrase, permeability, and efflux. Among 20 fluoroquinolones, compounds with a 3-methyl 3,7-diazabicyclo[3.3.0]octan-1(5)-ene-7-yl substituent at the C-7 position or a chlorine substituent at the C-8 position showed a good inhibitory activity on DNA gyrase (especially a mutated DNA gyrase). Compounds with a 3,7-diazabicyclo [3.3.0]octan-1(5)-ene-7-yl substituent at the C-7 position showed good permeability in the susceptible and resistant strains, while compounds with a fluorine substituent at the C-8 position were less effluxed from cells. PMID- 9875417 TI - Effects of chronic lead exposure on glutamate release and uptake in cerebellar cells of rat pups. AB - Changes in the release and uptake of glutamate in cerebellar granule and glial cells of offspring of lead-exposed mothers were determined. In cultured cerebellar granule cells exposed to lead for 5 days, glutamate release was less influenced upon N-methyl-D-aspartate (NMDA) stimulation than that in the control. Although the NMDA-stimulated release of glutamate in cerebellar granule cells prepared from lead-exposed first generation pups was not different from that of the control group, the S-nitroso-N-acetylpenicillamine (SNAP)-stimulated release of glutamate in cerebellar granule cells obtained from lead-treated pups was less elevated than that in the control. Furthermore, in cerebellar granule cells obtained from lead-exposed second generations pups, glutamate release did not respond to both NMDA and SNAP stimulation. In cerebellar glial cells exposed to lead, the basal glutamate uptake was not changed. However, the L-trans pyrollidine-2,4-dicarboxylic acid (PDC)-blocking effects was significantly reduced. In glial cells obtained from lead-exposed pups, the glutamate uptake was also less blocked by PDC than that in the control. Further decreases in PDC blocking effects were observed in cerebellar glial cells obtained from lead treated second generation pups compared to those from the control group. These results indicate that lead exposure induces the changes in the sensitivities of the glutamate release and uptake transporter. In addition, these results suggest that lead exposure might affect the intracellular signalling pathway and transmission in glutamatergic nervous system. PMID- 9875418 TI - Effects of the peroxisome proliferator ciprofibrate and prostaglandin F2 alpha combination treatment on second messengers in cultured rat hepatocytes. AB - Peroxisome proliferators induce hepatic peroxisome proliferation and hepatic tumors in rodents. These chemicals increase the expression of the peroxisomal beta-oxidation pathway and the cytochrome P-450 4A family, which metabolizes lipids, including eicosanoids. Peroxisome proliferators transiently induce increased cell proliferation in vivo. However, peroxisome proliferators are weakly mitogenic and are not co-mitogenic with epidermal growth factor (EGF) in cultured hepatocytes. Earlier study found that the peroxisome proliferator ciprofibrate is comitogenic with eicosanoids. In order to study possible mechanisms of the comitogenicity of peroxisome proliferator ciprofibrate and eicosanoids, we hypothesized that the co-mitogenicity may result from synergistic or additive increases of second messengers in mitogenic signal pathways. We therefore examined the effect of the peroxisome proliferator ciprofibrate, prostaglandin F2 alpha (PGF2 alpha) and the combination of ciprofibrate and PGF2 alpha with or without growth factors on the protein kinase C (PKC) activity, and inositol-1, 4, 5-triphosphate (IP3) and intracellular calcium ([Ca2+]i) concentrations in cultured rat hepatocytes. The combination of ciprofibrate and PGF2 alpha significantly increased particulate PKC activity. The combination of ciprofibrate and PGF2 alpha also significantly increased EGF, transforming growth factor-alpha (TGF-alpha) and hepatic growth factor (HGF)-induced particulate PKC activity. The combination of ciprofibrate and PGF2 alpha greatly increased [Ca2+]i. However, the increases of PKC activity and [Ca2+]i by ciprofibrate and PGF2 alpha alone were much smaller. Neither ciprofibrate or PGF2 alpha alone nor the combination of ciprofibrate and PGF2 alpha significantly increased the formation of IP3. The combination of ciprofibrate and PGF2 alpha, however, blocked the inhibitory effect of TGF-beta on particulate PKC activity and formation of IP3 induced by EGF. These results show that co-mitogenicity of the peroxisome proliferator ciprofibrate and eicosanoids may result from the increase in particulate PKC activity and intracellular calcium concentration but not from the formation of IP3. PMID- 9875419 TI - Nitric oxide synthase from bovine pancreas: purification and characterization. AB - Nitric oxide synthase, NOS (EC.1.14.13.39), was purified from bovine pancreas over 5,500-fold with a 7.6% yield using 30% ammonium sulfate precipitation, and 2',5'-ADP-agarose and calmodulin-agarose affinity chromatography. The purified bovine pancreatic NOS (bpNOS) showed a single band on SDS-PAGE corresponding to an apparent molecular mass of 160 kDa, whereas it was 320 kDa on non-denaturating gel-filtration. This indicated a homodimeric nature of the enzyme. The specific activity of the purified bpNOS was 31.67 nmol L-citrulline fored/mtn/mg protein and apparent K(m) for L-arginine was 15.72 microM. The enzyme activity was dependent on Ca2+ and calmodulin, and to a lesser extent on NADPH, FAD and FMN. H4B was not required as a cofactor for the activity. In an inhibition experiment with L-arginine analogues, NG-nitro-L-arginine (NNA) had the most potent inhibitory effect on bpNOS, and NG, NG'-dimethyl-L-arginine (symmetric; sDMA) did not have any inhibitory effect. Immunohistochemical analysis of the bovine pancreas using brain type NOS antibody (anti-bNOS antibody) revealed that acinar cells showed strong immunoreactivity against the antibody. PMID- 9875420 TI - Production of antibody against saikosaponin a, an active component of bupleuri radix. AB - High titer rabbit polyclonal antibodies (pAbs) which show a specificity for saikosaponin a (SSA), have been generated. The immunogen used was a conjugate of SSA linked through its glucose moiety to bovine serum albumin by periodate oxidation method. The antibody titers obtained from two rabbits, inoculated with the immunogen, reached a plateau after the fourth and third booster injection, respectively. The specificity of the pAbs was determined by hapten inhibition assays using several SSA-like structures. SSA competitively inhibited the binding of the rabbit anti-SSA pAbs to SSA-ovalbumin on solid phase, a coated antigen on the well. The antibodies showed high specificity to SSA, exhibiting no significant cross-reactivity with any of SSA analogues tested. PMID- 9875421 TI - Brazilin inhibits activities of protein kinase C and insulin receptor serine kinase in rat liver. AB - Hypoglycemic action of brazilin was found to be based on the improvement of peripheral glucose utility, and this action might be correlated with the insulin action pathway. In the present study we investigated the effect of brazilin on the insulin receptor autophosphorylation, protein kinase C (PKC), protein phosphatase and insulin receptor serine kinase in order to confirm whether the hypoglycemic mechanism is concerned with insulin action pathway. Brazilin was found to inhibit PKC and insulin receptor serine kinase, which are involved in the regulation of insulin signal pathway. But any significant effect was not shown on insulin receptor tyrosine kinase activity, autophosphorylation and phosphatase activity. These findings suggest that brazilin might enhance insulin receptor function by decreasing serine phosphorylation, which might mediate hypoglycemic effect of brazilin. PMID- 9875422 TI - Synthesis and in vitro cytotoxicity of cinnamaldehydes to human solid tumor cells. AB - Cinnamaldehydes and related compounds were synthesized from various cinnamic acids based on the 2'-hydroxycinnamaldehyde isolated from the bark of Cinnamomum cassia Blume. The cytotoxicity to human solid tumor cells such as A549, SK-OV-3, SK-MEL-2, XF498 and HCT15 were measured. Cinnamic acid, cinnamates and cinnamyl alcohols did not show any cytotoxicity against the human tumor cells. Cinnamaldehydes and related compounds were resistant to A549 cell line up to 15 micrograms/ml. In contrast, HCT15 and SK-MEL-2 cells were much sensitive to these cinnamaldehyde analogues which showed ED50 values 0.63-8.1 micrograms/ml. Cytotoxicity of the saturated aldehydes was much weak compared to their unsaturated aldehydes. From these studies, it was found that the key functional group of the cinnamaldehyde-related compounds in the antitumor activity is the propenal group. PMID- 9875423 TI - Cytotoxic constituents of Saussurea lappa. AB - The crude extract of Saussurea lappa displayed significant lethality to brine shrimp larvae. Investigation of the causative components by bioactivity-directed fractionation resulted in the isolation of three C17-polyene alcohols. Based on various nmr spectral data, these compounds were identified as shikokiols which had been previously isolated from Cirsium nipponicum and/or Centaurea aegyptica. These C17-polyene alcohols exhibited moderate cytotoxicities against the human tumor cell lines, A549, SK-OV-3, SK-MEL-2, XF498, and HCT15. PMID- 9875424 TI - Synthesis of antineoplaston A10 analogs as potential antitumor agents. AB - Several aniline mustard analogues were obtained by introducing N,N-bis(2 chloroethyl)amino moiety to phenyl ring of A10 analogues in order to increase reactivity of A10 analogs and selectivity into DNA. The in vitro antitumor activity of synthesized compounds was evaluated using five different solid tumor cell lines by SRB method. Aniline mustard analogues exhibited more potent antitumor activity than A10 analogs. Especially, m-aniline mustard of benzoyl analogue displayed remarkable antitumor activity. PMID- 9875425 TI - Conformationally constrained analogues of diacylglycerol having a perhydrofuro[3,4-c]furan-1,4-dione bis-gamma-butyrolactone skeleton. AB - Bis-gamma-lactones (1, 2) having a perhydrofuro[3,4-c]furan-1,4-dione skeleton were designed as conformationally constrained diacylglycerol analogues. They were synthesized from D-apiose in 11 steps, and evaluated as PKC-alpha ligands by measuring their ability to displace bound [3H]-PDBU from the enzyme. The compounds showed moderate binding affinities with Ki values of 13.89 (+/- 5.67) microM and 11.47 (+/- 0.89) microM, respectively. Their similar binding affinities indicate that these two bicyclic compounds were not effectively discriminated by PKC-alpha in terms of the direction of the side chain as other ligands built on similar bis-gamma-lactones. PMID- 9875426 TI - Epoxyalkanoyls as novel ACE inhibitors. AB - The epoxyalkanoyl derivatives were designed and synthesized as ACE inhibitors. Coupling of unsaturated carboxylic acids with amino acids and following epoxidation with dimethyldioxirane gave the epoxyalkanoyls with high yield. The inhibitory activity of synthesized compounds on angiotensin converting enzyme was IC50 values of 0.06-5.5 microM. PMID- 9875427 TI - Synthesis and evaluation of 5-aminosalicyl-glycine as a potential colon-specific prodrug of 5-aminosalicylic acid. AB - As a new colon-specific prodrug of 5-aminosalicylic acid (5-ASA), 5-aminosalicyl glycine (5-ASA-Gly) was prepared by a simple synthetic route in good yield. Apparent partition coefficients of 5-ASA-Gly were lower than those of 5-ASA, which determined in CHCl3/pH 6.8 buffer or n-octanol/pH 6.8 buffer system. Stability of 5-ASA-Gly by peptidases was investigated by incubation of 5-ASA-Gly with the homogenates of tissue and contents of stomach, proximal small intestine or distal small intestine of rats at 37 degrees C. 5-ASA was not detected, indicating that the prodrug was stable in the upper intestine. The amount of 5 ASA liberated from incubation of the prodrug in cecal or colonic contents of rats was about 65% or 27% in 8 hrs, respectively, which indicated that the prodrug activation took place more readily in the rat cecum whose bacterial counts are high like human colon. Results from in vitro experiments suggested 5-ASA-Gly as a promising candidate of a colon-specific prodrug of 5-ASA. PMID- 9875428 TI - Synthesis and properties of dextran-5-aminosalicylic acid ester as a potential colon-specific prodrug of 5-aminosalicylic acid. AB - Dextran-5-aminosalicylic acid ester (dextran-5-ASA) was synthesized as a colon specific prodrug of 5-aminosalicylic acid (5-ASA) which is active against inflammatory bowel diseases. Chemical stability of dextran-5-ASA in the bath of pH 1.2 or 6.8 was investigated at 37 degrees C for 6 hrs, and 5-ASA was not released on such conditions. Depolymerization (%) of dextran-5-ASA by dextranase with the degree of substitution (DS) of 18, 23, or 30 was 92, 62 or 45 in 8 hrs respectively, but was not affected by the MW of dextran (9,000, 40,600, 80,200 or 580,000). Distribution of 5-ASA in dextran, determined by gel filtration chromatography, appeared to be relatively uniform. Incubation of dextran-5-ASA (DS 18) in cecal contents of rats released 20% (28 g) and 35% (49 g) of 5-ASA in 8 hrs and 24 hrs, respectively, but no 5-ASA was liberated from small intestinal contents. PMID- 9875429 TI - Synthesis of the 7,8-dihydro-7-deazapurine derivatives and their antibiotic activity. AB - The cis- and trans-diastereomers of the 7,8-dihydro-7-deazapurine derivatives were synthesized from the corresponding diastereomers of 4-trans-cyano-2-methyl-3 phenyl-5-oxopyrrolidine (5), which were reduced from the 2-cis- and 2-trans diastereomers of 4-trans-cyano-2-hydroxymethyl-3-phenyl-5-oxopyrrolidine (2) via tosylation, iodination and following elimination, respectively. The prepared cis- and trans-diastereomers of 6-amino-2-mercapto-8-methyl-7-phenyl-7,8-dihydro-7(9H) deazapurine (8) were transferred to the corresponding 2-methylthio-diastereomers 9 and following desulfurization with Raney-nickel leaded to the cis- and trans diastereomers of 6-amino-8-methyl-7-phenyl-7,8-dihydro-7(9H)-deazapurine (10), respectively. The synthesized 7-deazapurine derivatives were tested for their antibiotic activity by the serial two-fold dilution method. PMID- 9875431 TI - 2-(1-Oxyalkyl)-1,4-dioxy-9,10-anthraquinones: synthesis and evaluation of antitumor activity. AB - Fourty eight derivatives of 2-(1-oxyalkyl)-1,4-dioxy-9,10-anthraquinone were synthesized, and their antitumor activity was evaluated. On the whole, 2-(1 hydroxyalkyl)-1,4-dihydroxy-9,10-anthraquinones (DHAQ = 1,4-dihydroxy-9,10 anthraquinone) showed stronger cytotoxic activity against L1210 cells than 2-(1 hydroxyalkyl)-1,4-dimethoxy-9,10-anthraquinones(DMAQ = 1,4-dimethoxy-9,10 anthraquinone), implying that free hydroxy groups at C-1 and C-4 of the anthraquinone structure are necessary for the cytotoxic activity. The bioactivity of 2-(1-hydroxyalkyl)-DHAQ derivatives differed according to the size of alkyl group at C-1; while the elongation of alkyl group over 7 carbon atoms failed to enhance the bioactivity, the derivatives possessing alkyl moiety of 1-6 carbon atoms showed an increase in the cytotoxicity and the antitumor activity in Sarcoma-180; 2-hydroxymethyl-DHAQ (ED50, 15 micrograms/ml; T/C, 125%), 2-(1 hydroxyethyl)-DHAQ(1.9 micrograms/ml; 139.2%), 2-(1-hydroxypropyl)-DHAQ (7.2 micrograms/ml; 135.1%), 2-(1-hydroxybutyl)-DHAQ (10.2 micrograms/ml; 125.3%), 2 (1-hydroxypentyl)-DHAQ (23.7 micrograms/ml; 110.1%), and 2-(1-hydroxyhexyl)-DHAQ (58 micrograms/ml; 108%). Next, 2-(1-Hydroxyalkyl)-DHAQ derivatives were acetylated to produce 2-(1-acetoxyalkyl)-DHAQ analogues. Although the acetylation somewhat enhanced the cytotoxicity, but not the antitumor action. In addition, the presence of phenyl group at C-1' enhanced the cytotoxicity and the T/C value, compared to alkyl groups of same size; 2-(1-hydroxy-1-phenyl)-DHAQ (ED50, 5.6 micrograms/ml; T/C, 137%). PMID- 9875430 TI - Synthesis and structure-activity relationship studies of substituted isoquinoline analogs as antitumor agent. AB - A number of substituted isoquinolin-1-ones, possible bioisosteres of the 5-aryl substituted 2,3-dihydroimidazo[2,1-a]isoquinolines, were synthesized and tested for their antitumor activity against five different human tumor cell lines. O-(3 hydroxypropyl) substituted compound (15) exhibited the best antitumor activity which is 3-5 times better than 5-[4'-(piperidinomethyl) phenyl]-2,3 dihydroimidazo[2,1-a]isoquinoline (1). PMID- 9875432 TI - Prolyl endopeptidase inhibitors from caryophylli flos. AB - Three prolyl endopeptidase inhibitors were isolated and identified as luteolin, quercetin and beta-sitosterol-3-O-beta-D-glucopyranoside with IC50 of 0.17, 0.19 and 27.5 ppm, respectively. The inhibition of two flavonoids were non-competitive with substrate. Twenty authentic flavonoids were tested in order to investigate structure-activity relationship. No significant relationship was found in them, however, catechol moiety of B-ring and 7-OH group in flavonoid skeleton were seemed to be responsible for the stronger activity. PMID- 9875433 TI - Coupled column chromatography in chiral separation of salmeterol. AB - A coupled achiral-chiral high-performance liquid chromatographic system has been developed for the determination of the enantiomers of salmeterol, S-(+) salmeterol and R-(-)-salmeterol in urine. The salmeterol was separated from the interfering components in urine and quantified on the silica column, and the enantiomeric composition was determined on a Sumichiral OA-4700 chiral stationary phase. The two columns were connected by a switching valve equipped with a silica precolumn. The precolumn was used to concentrate the salmeterol in the eluent from the achiral column before backflushing onto the chiral phase. The coupled system was validated. PMID- 9875434 TI - Resolution of salbutamol enantiomers in human urine by reversed-phase high performance liquid chromatography after derivatization with 2,3,4,6-tetra-O acetyl-beta-D-glucopyranosyl isothiocyanate. AB - A stereospecific HPLC method has been developed for the resolution of the enantiomers of salbutamol in human urine. After solid-phase extraction and derivatization with 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate, the diastereomeric derivatives were resolved (Rs = 1. 83) on 5 microns octadecylsilan column using 35% acetonitrile in 0.05 M ammonium acetate buffer (pH = 6) as a mobile phase with electrochemical detection. The diastereomeric derivatives were formed within 30 min. The detection limit of each enantiomer was 20 ng/ml (S/N = 3). PMID- 9875435 TI - Antihepatotoxic potential of Trianthema portulacastrum in carbon tetrachloride induced chronic hepatocellular injury in mice: reflection in haematological, histological and biochemical characteristics. AB - The effect of an ethanolic extract of the plant Trianthema portulacastrum L. on the CCl4-induced chronic hepatocellular damage of Swiss albino mice has been investigated. The normal mice received olive oil (0.2 ml/mouse) for five weeks. The CCl4 control mice, on the other hand, received CCl4 (0.05 ml/mouse) in olive oil for five weeks. The extract was administered at the dose of 100 mg/kg or 150 mg/kg for five weeks by gastric intubation in addition to CCl4 treatment. The CCl4 administration alone caused hepatocellular necrosis, severe anemia, leucopaenia, lymphocytopaenia, neutrophilia, eosinophilia and haemoglobinaemia along with the alterations of plasma albumin and globulin. The administration of plant extract (at 100 or 150 mg/kg) restored the CCl4-induced alterations of the haematological parameters to the normal level. The extract of T. portulacastrum elicited a marked protection against CCl4-induced hepatotoxicity as indicated by the several haematological parameters, related indices of formed elements, and different fractions of plasma protein. We also observed the dose-dependent antihepatotoxic effect of the extraction on these mice. The 150 mg/kg of extract was found to be more effective in normalizing the toxic effects of CCl4 on the above parameters of mice. These results suggest that the hepatoprotective effect of T. portulacastrum could be caused by its critical involvement in modulating several factors associated with erythropoiesis, and the boosting of general immunity of the host. PMID- 9875436 TI - A biotin-avidin labeled enzyme immunoassay for the quantitation of serum TSH using protein-layered solid phase. AB - A sensitive enzyme immunoassay for serum TSH has been developed utilizing the tight binding between biotin and avidin, and three layered protein polystyrene beads as solid phase. To increase binding capacity of TSH and sensitivity of the assay, the polystyrene beads were coated sequentially with mouse immunoglobulin as first layer, rabbit antimouse immunoglobulin as second layer and monoclonal anti-TSH as third layer. A serum sample was incubated simultaneously with a monoclonal anti-TSH immobilized polystyrene beads and a second monoclonal anti TSH covalently attached to biotin. After washing, the antibody bound serum TSH anti-TSH-biotin complex is reacted with horseradish peroxidase (HRP)-labeled avidin. Following a second wash, the bound HRP activity was measured colorimetrically. Reproducible results were obtained within 4 hours for serum TSH in the range between 0 microIU/ml and 50 microIU/ml with detection limit of 0.1 microIU per test. PMID- 9875437 TI - Role of the hepatic xanthine oxidase in thyroid dysfunction: effect of thyroid hormones in oxidative stress in rat liver. AB - The effect of thyroid hormones on the hepatic xanthine oxidase activity was studied in rats after the intraperitoneal injections of comthyroid (triiodotyronine:thyroxine = 1:4) at 0.3 mg/kg for 3 consecutive days. The aim of this study was to understand the precise mechanism of hyperthyroidism induced by oxidative stress. The concentration of lipid peroxides determined indirectly by the measurement of thiobarbituric acid reactants was increased in comthyroid treated rats. The hepatic glutathione content was decreased in comthyroid injected rat compared to the euthyroid state. It was also observed that the increment of xanthine oxidase activity has a profound role in oxygen radicals generation system in comthyroid treated rat. These findings suggest that the enhanced xanthine oxidase activity and depleting glutathione content in comthyroid treated rats result in pathophysiological oxidative stress including an increment of hepatic lipid peroxidation. PMID- 9875439 TI - Prolonged systemic delivery of streptokinase using liposome. AB - To prolong the biological half-life of streptokinase, a thrombolytic agent, streptokinase-bearing liposome with and distearolyphosphatidyl ethanolamine-N poly (ethylene glycol) 2000 (DSPE-PEG 2000) was prepared and evaluated. Streptokinase-bearing liposomes composed of distearolyphosphatidylcholine (DSPC), cholesterol and cholesterol-3-sulfate with DSPE-PEG 2000 was prepared by the freeze-thawing method and administered via femoral vein to rats (15,000 IU/kg). The activity of streptokinase in plasma was determined by the method based on the amidolytic activity of streptokinase-plasminogen complex. Pharmacokinetic parameters of streptokinase incorporated in liposomes were compared with those of streptokinase alone. The T1/2 and AUCinfinity of streptokinase incorporated in DSPC-PEG liposome increased 16.3- and 6.1-fold, respectively, compared with those of streptokinase alone. Streptokinase-bearing long-circulating liposome could increase the circulation time of streptokinase in blood and expect longer thrombolytic activity compared with streptokinase alone. PMID- 9875438 TI - Biochemical and pharmacological properties of a new proton pump inhibitor, 2 amino-4,5-dihydropyrido[1,2-a]thiazolo [5,4-g] benzimidazole (YJA20379-5). AB - This study was designed to determine biochemical and pharmacological properties of a newly synthesized benzimidazole derivative, 2-amino-4,5-dihydropyrido [1,2 a] thiazolo [5,4-g] benzimidazole (YJA20379-5) in vitro and in vivo. In the leaky membrane vesicles of pig gastric mucosa, YJA20379-5 inhibited the K(+)-stimulated H+,K(+)-ATPase activity in a concentration- and time-dependent manner, with IC50 values being 43 microM and 31 microM at pH 6.4 and 7.4, respectively. YJA20379-5, given intraduodenally, had a potent inhibitory effect on the gastric acid secretion in pylorus-ligated rats. The ED50 value for acid secretion was 15.4 mg/kg. YJA20379-5, administered orally, also suppressed gastric damages induced by water-immersion stress, indomethacin and ethanol, and duodenal damage induced by mepirizole in rats; the ED50 values were 17.6, 4.7, 3.0 and 18.7 mg/kg, respectively. Furthermore, repeated oral administration of YJA20379-5 accelerated the spontaneous healing of acetic acid-induced gastric ulcers in rats. It is concluded that the antisecretory activity of YJA20379-5 appears to be associated with inhibition of H+,K(+)-ATPase, while its antigastric and antiduodenal lesion activities are primarily related to the antisecretory effect. PMID- 9875440 TI - Comparison between ELISA and gel-filtration assay for the quantitation of airway mucins. AB - In this study, we developed immunoassay methods for the more convenient and effective detection of rat tracheal mucin and the results were compared with those of [3H]glucosamine based gel-filtration method. A monoclonal anti-rat tracheal mucin antibody, mAbRT03, which specifically recognizes rat tracheal mucins, was used throughout in this study. To induce mucin secretion, varying concentrations of ATP (0-2 mM) were applied to the primary rat tracheal surface epithelial (RTSE) cell culture which had been metabolically radiolabeled with [3H]glucosamine and the secretion of mucin was analyzed both by the immunoassay and the gel-filtration chromatography methods. For the immunoassay, the following two procedures were employed. 1) Simple ELISA; the culture spent media were directly coated onto the assay plate and the immunoreactivity with mAbRT03 was assessed from the standard curve generated with the purified rat mucin. 2) Inhibition ELISA; A known amount of the purified rat mucin was coated onto the assay plate and then ATP-stimulated culture spent media were added to inhibit the immunoreactivity with mAbRT03. The contents of mucin in the sample were calculated from the standard inhibition curve which was generated with the purified rat mucin. The assay results obtained from the immunoassays were identical with those from the gel-filtration methods. The present result indicates that ELISA can be substituted for the laborious, time-consuming gel filtration assay in studying the regulation of airway mucin release from cultured airway epithelial cells. PMID- 9875441 TI - In vitro angiogenic activity of Aloe vera gel on calf pulmonary artery endothelial (CPAE) cells. AB - Angiogenic activity of Aloe vera gel was investigated by in vitro assay. We obtained the most active fraction from dichloromethane extract of Aloe vera gel by partitioning between hexane and 90% aqueous methanol. The most active fraction (F3) increased the proliferation of calf pulmonary artery endothelial (CPAE) cells. In addition, F3 fraction induced CPAE cells to invade type 1 collagen gel and form capillary-like tube through in vitro angiogenesis assay, and increased the invasion of CPAE cells into matrigel through in vitro invasion assay. Furthermore, the effect on the mRNA expression of proteolytic enzymes which are key participants in the regulation of extracellular matrix degradation was investigated by northern blot analysis. F3 fraction enhanced mRNA expression of urokinase-type plasminogen activator (u-PA), matrix metalloproteinase-2 (MMP-2), and membrane-type MMP (MT-MMP) in CPAE cells whereas the expression of plasminogen activator inhibitor-1 (PAI-1) mRNA was not changed. PMID- 9875442 TI - Antitumor activity of the aqueous-alcoholic extracts from unripe cotton ball of Gossypium indicum. AB - The present study investigated the antitumor activity of the aqueous-alcoholic extracts from unripe cotton balls of Gossypium indicum. An Exposure of murine B16 melanoma and L1210 lymphoma cells to the extracts resulted in their severe deaths in time- and concentration-dependent manners. Of the extracts, hydrophilic fractions were most efficacious for the antitumor activity and found to contain certain amounts of catechin and its derivatives. The hydrophilic extract fraction C36B2-8 had approximately 10 times more cytotoxic effects on B12 and L1210 cells than on isolated murine thymocytes. High concentrations (> 150 micrograms/ml) of C 36B3-8 mainly induced necrotic cell death. At low concentrations (< 100 micrograms/ml), however, C 36B3-8 induced not only necrosis but also apoptosis of the two tumor cell lines, which was proved by the TUNEL staining and DNA fragmentation techniques. The data indicate that certain ingredients of the cotton ball extract of G. indicum have an antitumor activity. PMID- 9875443 TI - Structure-activity relationships of gagaminine and its derivatives on the inhibition of hepatic aldehyde oxidase activity and lipid peroxidation. AB - In order to determine the structure-activity relationships for antioxidative effects of gagaminine, a steroidal alkaloid isolated from the roots of Cynanchum wilfordi (Asclepiadaceae), two derivatives identified as sarcostin and penupogenin were prepared from gagaminine by hydrolysis and reduction. These compounds were evaluated for the inhibitory effects on the aldehyde oxidase activity and on lipid peroxidation in vitro. Furthermore, their effects were compared with those of gagaminine and the related compounds, cinnamic acid and nicotinic acid. The results of this study prove that the cinnamoyl group in the structure of gagaminine is critical in inhibition of the aldehyde oxidase activity while the nicotinoyl group may be necessary for anti-lipid peroxidation of the compound. PMID- 9875444 TI - Screening of new bioactive materials from microbial extracts of soil microorganism (I). Antimicrobial activity from 200 samples using microdilution assay. AB - The microdilution assay recommended by NCCLS (National Committee for Clinical Laboratory Standards) is one of the standardized methods of antibiotic susceptibility test. This method has been widely used clinically to obtain MIC values of antibiotics on pathogenic microorganisms. It is more convenient, rapid and simple to test many samples than other test methods such as agar diffusion assay and broth macrodilution assay. The screening of antimicrobial agents from microbial extracts is too laborious in its process. Therefore, a number of screening methods having more simple procedure have been developed. In our laboratory, we applied microdilution assay for screening the antimicrobial agents. This assay showed dose-response results and was more sensitive than disc diffusion assay in our system. We tested 200 samples of microbial extracts originated from 100 microbial strains and selected several samples as potential candidates. In this report, we show that the microdilution assay is more convenient method in screening of antibiotic susceptibility than those previously reported. PMID- 9875445 TI - Demographic characteristics of zipeprol-associated deaths in Korea. AB - The abuse of zipeprol, an antitussive agent, was found to be most prevalent among young people in Korea. Because abusers take large doses of this drug for its hallucinogenic effects, fatalities from zipeprol overdose abuse have been on the rise since 1991. Since 1991, a total of 69 zipeprol-related deaths have occurred throughout the nation. A demographic study shows that in ninety six percent of cases involving ziperol alone, the victims were in their teens and twenties. The male/female ratio in zipeprol related death was 3.5:1. Most of these zipeprol associated deaths occurred in the larger cities of Seoul and Inchon. The blood concentration of zipeprol ranged from 0.8 to 38.3 micrograms/mL in single drug involved deaths, while zipeprol varied from 0.1 to 35.3 micrograms/mL in zipeprol and dextromethorphan victims. PMID- 9875446 TI - Purification and characterization of Cop, a protein involved in the copy number control of plasmid pE194. AB - Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCl buffer (pH 7.5) containing 100 mM NaCl. Cop protein was calculated to contain 39.1% alpha helix, 16.8% beta-sheet, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl aminopeptidase in E. coli. PMID- 9875447 TI - All-trans retinoic acid induced differentiation of rat mammary epithelial cells cultured in serum-free medium. AB - Retinoids are applied to not only cancer prevention but also cancer chemotherapy by stimulating differentiation of cells. We studied differentiation inducing effect of all-trans retinoic acid (ATRA) by studying proportion of high dense fractions of stem-like cells and the size of S phase fraction in cell cycle. From mammary organoids obtained from 7- to 8-week old F344 female rat mammary gland, we cultured rat mammary epithelial cells (RMEC) and treated physiological doses of 10(-6), 10(-7), and 10(-8) M ATRA from the first day and then cultured for 4, 7, and 14 days. After that, immunostaining was performed using peanut agglutinin (PNA) and anti-Thy-1.1 monoclonal antibody (Thy-1.1) that can be used as markers of differentiation. We separated four different cell subpopulations by flow cytometry: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy 1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). We observed continuous decreases of high dense fractions of stem-like cells (PNA+ subpopulations) for 14 days and as much decreases as high doses of ATRA, which were thought to be proportional to doses of ATRA. We labeled RMEC with bromodeoxyuridine and investigated cell cycle fractions that went through S phase. We observed a tendency of decrease of S phase fraction with time in culture, which is thought to be related to continuous decreases of PNA+ subpopulations and inhibitory role of ATRA on cell cycle. These results suggest that physiological doses of ATRA could stimulate differentiation of RMEC and convert stem-like RMEC to differentiated cells in SFM for a relatively long period of 14 days. PMID- 9875448 TI - Expression of recombinant human cytochrome P450 1A2 in Escherichia coli bacterial mutagenicity tester strain. AB - Human cytochrome P450 1A2 is one of the major cytochrome P450s in human liver. It is known to be capable of activating a number of carcinogens such as arylamines and heterocyclic amines. In order to develop the new bacterial mutagenicity test system with human P450, a full length of human P450 1A2 cDNA inserted into pCW bacterial expression vector was introduced to Escherichia coli WP2 uvrA strain which is a well-known E. coli strain for bacterial reverse mutagenicity assay. Expressed human P450 1A2 showed typical P450 hemoprotein spectra. Maximum expression was achieved at 48 hrs after incubating at 30 degrees C in terrific broth containing ampicillin, IPTG and other supplements. High level expression of P 450 1A2 in E. coli WP2 uvrA membranes was determined in SDS-PAGE. The well known mutagens 2-aminoanthracene and MelQ increased the revertant colonies of E. coli WP2 uvrA expressing human P450 1A2 without an exogenous rat hepatic post mitochondrial supernatant (S9 fraction) in a dose-dependent manner. The results show that the functional expression of human P450 in bacterial mutagenicity tester strain will provide a useful tool for studying the mechanism of the mutagenesis and carcinogenesis of new drugs and environmental chemicals. PMID- 9875449 TI - Resistance mechanism of Acinetobacter spp. strains resistant to DW-116, a new quinolone. AB - DW-116 is a new fluoroquinolone antimicrobial agent with a broad spectrum. In order to elucidate the resistance mechanism to DW-116 in Acinetobacter spp. bacteria, total chromosomal DNA was isolated from 10 strains of Acinetobacter spp. resistant to DW-116. Quinolone resistance determinant region (QRDR) of DNA gyrase gene was amplified by PCR. The 345 bp nucleotide fragment yielded was inserted into pKF 3 which was used as the vector. Comparisons of the DNA sequences of 8 strains with that of the wild type strain revealed a Ser-83 to Leu mutation in mutants and all ten strains contained one silent mutation(T-->G) in QRDR. From Acinetobacter MB4-8 strain, DNA gyrase was isolated and purified, through no-vobiocin-sepharose, heparin-sepharose affinity column chromatography. The enzyme was composed of two subunits and the molecular mass of subunits A and B were 75.6 and 51.9 kDa, respectively. The supercoiling activity of the reconstituted DNA gyrase composed of subunit A from Acinetobacter MB4-8 and subunit B from E. coli was not inhibited by 128 micrograms/ml of ciprofloxacin. It might be said that one of the resistance mechanisms to DW-116 in A cinetobacter MB4-8 was subunit A alteration of DNA gyrase. PMID- 9875450 TI - The involvement of protein kinase C and tyrosine kinase in vanadate-induced contraction. AB - Gastric smooth muscle of cats was used to investigate the involvement of protein kinase in vanadate-induced contraction. Vanadate caused a contraction of cat gastric smooth muscle in a dose-dependent manner. Vanadate-induced contraction was totally inhibited by 2 mM EGTA and 1.5 mM LaCl3 and significantly inhibited by 10 microM verapamil and 1 microM nifedipine, suggesting that vanadate-induced contraction is dependent on the extracellular Ca2+ concentration. and the influx of extracellular Ca2+ was mediated through voltage-dependent Ca2+ channel. Both protein kinase C inhibitor and tyrosine kinase inhibitor significantly inhibited the vanadate-induced contraction and the combined inhibitory effect of two protein kinase inhibitors was greater than that of each one. But calmodulin antagonists did not have any influence on the vanadate-induced contraction. On the other hand, both forskolin (1 microM) and sodium nitroprusside (1 microM) significantly inhibited vanadate-induced contraction. Therefore, these results suggest that both protein kinase C and tyrosine kinase are involved in the vanadate-induced contraction which required the influx of extracellular Ca2+ in cat gastric smooth muscle, and that the contractile mechanism of vanadate may be different from that of agonist binding to its specific receptor. PMID- 9875451 TI - Characterization of tTA and its functional domain in tetracycline repressor mediated gene repression system. AB - To elucidate of role(s) of tTA as a repressor in the tTA-mediated gene repression system, we introduced mutations into the acidic domain of VP16 and examined the effects of such various mutations. In the transient repression experiment, a region containing 34 amino acids of the activation domain of VP16 (412-456) which interacts with TFIIB was found to be necessary and sufficient for the tTA mediated repression of gene expression. However, in the experiment to investigate the fact that tTA-regulated repression is related to the activation function of VP 16, we found that the repression abilities of tTA derivatives did not correlate exactly with their activation abilities. Therefore, we conclude that increased mass of VP16 in tTA might be also important for efficient repression in addition to functional domain of VP16. PMID- 9875452 TI - Studies on the synthesis of naphthoquinoids. AB - Four derivatives of 6-oxo-3,4,4a,5-tetrahydro-3-hydroxy-2,2-dimethylnaphtho-1,2 pyran (1), known as bactericidal, bacteriostatic, fungicidal, fungistatic agents, were synthesized to investigate the effect of substituents on the aromatic ring. PMID- 9875453 TI - Reactivity and suitability of t-Boc-protected thiophosphotyrosine intermediate analogs for the solid or solution phase peptide synthesis. AB - N-(tert-Butoxycarbonyl)-O-(dimethylthiophosphono)-L-tyrosine (6) and N-(tert butoxycarbony)-O-(dicyanoethylthiophosphono)-L-tyros ine (15) were prepared as intermediates for the synthesis of thiophosphotyrosine-containing peptides. The reactivity and suitability of two compounds for the solid phase or solution phase peptide synthesis utilizing t-Boc chemistry were examined. PMID- 9875454 TI - Synthesis and antiviral activity of 2'-fluorohexopyranosyl nucleosides. AB - 2'-Fluorohexopyranosyl nucleosides 1a and 1b which contained a bioisosteric double bond and a fluorine were synthesized in 12 steps, starting from D galactose. During diethylaminosulfur trifluoride (DAST) fluorination, retention of stereochemistry was observed through the participation of methoxy or chloro group at the 6-position of the purine base. The final nucleosides 1a and 1b were found to be inactive against HIV-1 and HSV-1,2. PMID- 9875455 TI - Isolation of a multidrug resistance inhibitor from Aconitum pseudo-laeve var. erectum. AB - To overcome multidrug resistance (MDR) in cancer chemotherapy, we prepared various plant extracts and searched for a component which is effective for inhibition of MDR. MDR inhibition activity was determined by measuring cytotoxicity to MDR cells using multidrug resistant human fibrocarcinoma KB V20C, which is resistant to 20 nM vincristine and expresses high level of mdr1 gene. Of various plant extracts, the MeOH extract of the root of Aconitum pseudo-laeve var. erectum was found to have potent inhibitory activity on MDR. The bioassay guided fractionation of the MeOH extract of the plant led to the isolation of an alkaloid, lycaconitine, as an active principle. And the IC50 of lycaconitine for KB V20C cells was 74 micrograms/ml. PMID- 9875456 TI - Antibacterial activity of Camellia sinensis extracts against dental caries. AB - Different bacteria were separated from saliva and teeth of cariogenic patients and identified by a variety of morphological and biochemical tests. Extracts of green tea strongly inhibited Escherichia coli, Streptococcus salivarius and Streptococcus mutans. The antibacterial effect of green and black tea extracts were compared with those of amoxicillin, cephradine and eugenol. PMID- 9875458 TI - Isolation of 3-O-(4'-hydroxybenzyl)-beta-sitosterol and 4-[4'-(4" hydroxybenzyloxy)benzyloxy]benzyl methyl ether from fresh tubers of Gastrodia elata. AB - Two new 4-hydroxybenzyl alcohol derivatives (1 and 2) were isolated from the methanol extract obtained from fresh tubers of Gastrodia elata together with 4 hydroxybenzyl methyl ether, 4-hydroxybenzyl alcohol, bis(4-hydroxyphenyl)methane, 4-hydroxybenzaldehyde, beta-sitosterol and palmitic acid. 1 and 2 were identified as 3-O-(4'-hydroxybenzyl)-beta-sitosterol and 4-[4'-(4" hydroxybenzyloxy)benzyloxy]benzyl methyl ether, respectively, according to the spectroscopic data. PMID- 9875457 TI - Boron trifluoride etherate on silica-A modified Lewis acid reagent (VII). Antitumor activity of cannabigerol against human oral epitheloid carcinoma cells. AB - Geraniol (1), olivetol (2), cannabinoids (3 and 4) and 5-fluorouracil (5) were tested for their growth inhibitory effects against human oral epitheloid carcinoma cell lines (KB) and NIH 3T3 fibroblasts using two different 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. Cannabigerol (3) exhibited the highest growth-inhibitory activity against the cancer cell lines. PMID- 9875459 TI - Synthesis of new prostaglandin F2 alpha derivatives. PMID- 9875460 TI - Differential effects of nongenotoxic and genotoxic carcinogens on the preneoplastic lesions in the rat liver. AB - Glutathione S-transferase placental form (GST-P) positive foci development and its expression in liver exposed by nongenotoxic carcinogens phenobarbital (PB) and clofibrate (CF), and genotoxic carcinogen 2-amino-3-methylimidazo[4,5-f] quinoline (IQ) were investigated as a measure of carcinogenic potential of these chemicals. Male F344 rats were initially given a single intraperitioneal injection of diethylnitrosamine (200 mg/kg), and 2 weeks later, animals were fed diets containing 0.03% IQ or 0.5% CF or 0.05% PB or basal diet as a control for 6 weeks. All rats were subjected to two-thirds partial hepatectomy (PH) at week 3. Sequential sacrifice of rats was performed at 8 weeks or 52 weeks, and liver tissues were examined for immunohistochemical staining of GST-P positive foci. The numbers (No./cm2) and areas (mm2/cm2) of GST-P positive foci were increased by IQ or PB, but were decreased by CF compare to the control. Consistent with the development of GST-P positive foci, a time-related increase in the expression of GST-P mRNA was found in the rats treated with IQ, whereas CF decreased it. The incidence of hepatocellular carcinoma at 52 weeks was increased by all three chemicals. These results show that PB and IQ induced GST-P positive foci, but the peroxisome proliferator CF did not, which suggest that the prediction of carcinogenic potency based on the development of prenoplastic foci may cause false negative in a particular category compounds like peroxisome proliferators. PMID- 9875461 TI - Studies on the effects of piperidine derivatives on blood pressure and smooth muscles contractions. AB - Ten substituted phenacyl derivatives of 4-hydroxypiperidine were synthesized and studied for their effects on the mean arterial blood pressure (MABP) in normotensive anaesthetized rats and smooth muscles contractions of isolated rabbit jejunum. Two derivatives caused fall in blood pressure at the dose of 10 20 mg/kg and one rise in blood pressure at the dose of 20 mg/kg. Two compounds exhibited biphasic response (hypotensive followed by hypertensive) and one gave triphasic response at 10 mg/kg dose. Rest of four derivatives were found devoid of any effect on mean arterial blood pressure up to the dose of 30 mg/kg. All the derivatives except two caused relaxant effect on the spontaneous contraction of rabbit jejunum at the dose range of 0.1-2 mg/kg. PMID- 9875462 TI - Dose dependency of earthworm powder on antithrombotic and fibrinolytic effects. AB - The freeze-dried powder of Lumbricus rubellus earthworm was administered orally to rats and its fibrinolytic and antithrombotic effects were investigated. The fibrinolytic activity of plasma was determined by measuring the plasmin activity of the euglobulin fraction and was increased to two-folds of the control at a dose of 0.5 g/kg/day and five times with 1 g/kg/day after 4-day administration. The antithrombotic effect was studied in an arterio-venous shunt model of rats. The thrombus weight decreased significantly from 43.2 mg to 32.4 mg at a dose of 0.5 g/kg/day after 8-day treatment. The level of fibrinogen/fibrin degradation product (FDP) in serum was elevated in a dose-dependent manner during the treatment period. On the 8th day after administration, the FDP value was increased to 7.7 micrograms/ml compared with the control value of 3.3 micrograms/ml. These results support that earthworm powder is valuable for the prevention and/or treatment of thrombotic conditions. PMID- 9875463 TI - Farnesylcysteine methyltransferase activity and Ras protein expression in human stomach tumor tissue. AB - The processing pathway of G-proteins and Ras family proteins includes the isoprenylation of the cysteine residue, followed by proteolysis of three terminal residues and alpha-carboxyl methyl esterification of the cysteine residue. Farnesylcysteine methyltransferase (FCMT) activity is responsible for the methylation reaction which play a role in the membrane attachment of a variety of cellular proteins. Four kinds of Ras protein (c-Ha-ras, c-N-Ras, c-Ki-Ras, pan Ras) expression were detected in adenocarcinoma of human tissue by immunohistochemical method, and hematoxylin and eosin staining. The level of Ras protein in human stomach tumor tissues was much higher than in normal and peritumoral regions of the same biopsy samples. The FCMT activities of each cellular fractions were high in mitochondrial fraction followed by microsomal fraction, whole homogenate and cytosolic fraction. The inhibitory effect on FCMT activity on stomach tumor tissue was determined after treatment with 0.25 microM of S-adenosyl-L-homocysteine. S-adenosyl-L-homocysteine inhibited FCMT activity from 11.2% to 30.5%. These results suggested that FCMT might be involved in Ras proteins activity. PMID- 9875464 TI - Sequence selectivity of DNA alkylation by adozelesin and carzelesin. AB - Adozelesin and carzelesin are synthetic analogues of the extremely potent antitumor antibiotic CC-1065, which alkylates N3 of adenine in a consensus sequence 5'-(A/T)(A/T)A* (A* is the site of alkylation). We have investigated the DNA sequence selectivity of adozelesin and carzelesin by thermally induced DNA strand cleavage assay using radiolabeled restriction DNA fragments. An analysis of alkylation patterns shows that the consensus sequences for carzelesin and adozelesin have been found to be 5'-(A/T)(A/T)A* and 5'-(A/T)(G/C)(A/T)A*. A new consensus sequence, 5'-(A/T)(A/T)CA*, has been observed to display an additional alkylation site for adozelesin but not for carzelesin. These results indicate that the pattern of sequence selectivity induced by carzelesin is similar but not identical to those induced by adozelesin. PMID- 9875465 TI - Genotoxicity study of bojungchisup-tang, an oriental herbal decoction-in vitro chromosome aberration assay in Chinese hamster lung cells and in vivo supravital staining micronucleus assay with mouse peripheral reticulocytes. AB - The toxicity evaluation of oriental herbal drugs is of great concern at present. Bojungchisuptang (BCST, in Korean), a decocted medicine of oriental herbal mixture, is now well used in clinic at oriental hospitals for the treatment of edema of several diseases in practice. However, the toxicity of the oriental herbal decocted medicines such as genetic toxicity is not well defined until now. In this respect, to clarify the genetic toxicity of BCST, in vitro chromosome aberration assay with Chinese hamster lung (CHL) fibroblasts and in vivo supravital micronucleus assay with mouse peripheral reticulocytes were performed in this study. In the chromosome aberration assay, we used 5,000 micrograms/ml BCST as maximum concentration because no remarkable cytotoxicity in CHL cells was observed both in the presence and absence of S-9 metabolic activation system. No statistical significant differences of chromosome aberrations were observed in CHL cells treated with 5,000, 2,500 and 1,250 micrograms/ml BCST for 6 hour both in the presence and absence of S-9 metabolic activation. However, very weak positive result (6.5-8.0% aberration) of BCST was obtained in the absence of S-9 metabolic activation system at 5,000 micrograms/ml BCST when treated for 24 hour, i.e. 1.5 normal cell cycle time. And also, in vivo clastogenicity of BCST was studied by acridine orange-supravital staining micronucleus assay using mouse peripheral reticulocytes. We used 2,000 mg/kg as the highest oral dose in this micronucleus assay because no acute oral toxicity of BCST was observed in mice. The optimum induction time of micronucleated reticulocytes (MNRETs) was determined as 36 hours after oral administration of 2,000 mg/kg BCST. No significant differences of MNRETs between control and BCST treatment groups were observed in vivo micronucleus assay. From these results, BCST revealed very weak positive result in chromosome aberration assay in vitro with CHL cells and no clastogenicity in micronucleus assay in vivo. PMID- 9875466 TI - Induction of differentiation of the cultured rat mammary epithelial cells by triterpene acids. AB - We investigated the effects of triterpene acids (TAs), ursolic acid (UA) and oleanolic acid (OA), on the induction of proliferation and differentiation of normal rat mammary epithelial cells (RMEC) or organoids cultured in Matrigel or primary culture system. To elucidate the effects, we tested their differentiation inducing activities with intercellular communication ability, cell cycle patterns, induction of apoptosis, and morphological differentiation in the three dimensional extracellular culture system. To study the changes of RMEC subpopulation in culture, the cultured cells were isolated, immunostained with peanut lectin (PNA) and anti-Thy-1.1 antibody and then analyzed with flow cytometry. Four different subpopulations, such as PNA and Thy-1.1 negative cells (B-), PNA positive cells (PNA+), Thy-1.1 positive cells (Thy-1.1+), PNA and Thy 1.1 positive cells (B+), were obtained and the size of each subpopulation was changed in culture with time in the presence of TAs. Intercellular communication was observed in culture for 7 days in TAs-treated cells, but not in culture for 4 days with scrape-loading dye transfer technique. G2/M phase cells and the number of apoptotic population were increased in TAs-treated groups in cell cycle analyses. S phase fractions were reduced and the change of G1 phase cells was not observed. The colonies with distinct multicellular structures, such as stellate, ductal, webbed, squamous, lobulo-ductal colonies, were observed in Matrigel culture and the frequencies of each colony were changed in the presence of TAs. These results suggest that UA and OA have differentiation inducing effects on rat mammary epithelial cells in primary or in Matrigel culture. PMID- 9875468 TI - No effect of diltiazem on the hepatic clearance of indocyanine green in the rats. AB - In order to investigate the effect of the pretreatment with various doses of diltiazem (DTZ) on the pharmacokinetics of indocyanine green (ICG) at steady state, especially the hepatic blood clearance due to the change of hepatic blood flow, the following experiments were carried out with ICG, a hepatic function test marker, not metabolized in liver and only excreted in bile. The intravenous bolus injection (3,780 micrograms/kg) and the constant-rate infusion (10,100 micrograms/kg/hr) of ICG into the left femoral vein were made in order to check the steady-state plasma concentration (Css of 10 micrograms/ml) of ICG at 20, 25 and 30 min. Following a 90-min washout period, the intravenous bolus injection (108, 430, 860 and 1,720 micrograms/kg) and the constant-rate infusion (108, 433, 866 and 1,730 micrograms/kg/hr) of DTZ into the right femoral vein were made and the achievement of the steady-state plasma levels (Css of 50, 200, 400 and 800 ng/ml) of DTZ were conformed at 60, 70 and 80 min. During the steady state of DTZ, the intravenous bolus injection (3,780 micrograms/kg) and the constant-rate infusion (10,200 micrograms/kg/hr) of ICG into the left femoral vein were made and also the steady-state plasma concentration of ICG was checked at 20, 25 and 30 min. The plasma concentrations of DTZ and ICG were determined using a high performance liquid chromatographic technique. At the steady state, the hepatic blood clearance of ICG was obtained from the plasma concentration and blood-to plasma concentration ratio (RB) of ICG. The pretreatment with various doses of DTZ did not influence the plasma concentrations, RB and plasma free fraction (fp) of ICG. So the hepatic blood clearance of ICG was independent of concentration of DTZ. The hepatic blood clearance of ICG could be affected by both hepatic blood flow and hepatic intrinsic clearance. But there was no change of the hepatic blood clearance of ICG between the control and the DTZ-pretreated rats in this study. So it may be suggested that DTZ does not influence hepatic blood flow. PMID- 9875469 TI - Clonazepam release from poly(DL-lactide-co-glycolide) nanoparticles prepared by dialysis method. AB - Aim of this work is to prepare poly(DL-lactide-co-glycolide) (PLGA) nanoparticles by dialysis method without surfactant and to investigate drug loading capacity and drug release. The size of PLGA nanoparticles was 269.9 +/- 118.7 nm in intensity average and the morphology of PLGA nanoparticles was spherical shape from the observation of SEM and TEM. In the effect of drug loading contents on the particle size distribution, PLGA nanoparticles were monomodal pattern with narrow size distribution in the empty and lower drug loading nanoparticles whereas bi- or trimodal pattern was showed in the higher drug loading ones. Release of clonazepam from PLGA nanoparticles with higher drug loading contents was slower than that with lower loading contents. PMID- 9875467 TI - Amentoflavone, a plant biflavone: a new potential anti-inflammatory agent. AB - Biflavonoid is one of unique classes of naturally-occurring bioflavonoids. Certain biflavonoids including amentoflavone were previously reported to have inhibitory effect on the group II phospholipase A2 activity. Amentoflavone was also found to inhibit cyclooxygenase from guinea-pig epidermis without affecting lipoxygenase. In this study, anti-inflammatory and analgesic activities of amentoflavone were evaluated. When amentoflavone was administered intraperitoneally, it showed a potent anti-inflammatory activity as determined by amelioration of croton-oil induced mouse ear edema. It also showed a potent anti inflammatory activity in the rat carrageenan paw edema model (ED50 = 42 mg/kg) compared to the activity of prednisolone (35 mg/kg) and indomethacin (10 mg/kg). However, amentoflavone did not show a significant inhibitory activity against rat adjuvant-induced arthritis, a chronic inflammatory model. In addition, amentoflavone was found to possess a potent analgesic activity in the acetic acid writhing test (ED50 = 9.6 mg/kg) compared to the activity of indomethacin (3.8 mg/kg). These results suggest that amentoflavone may be a potential lead for a new type of anti-inflammatory agents having dual inhibitory activity of group II phospholipase A2 and cyclooxygenase. PMID- 9875470 TI - Differential coupling of G alpha q family of G-protein to muscarinic M1 receptor and neurokinin-2 receptor. AB - The ligand binding signals to a wide variety of seven transmembrane cell surface receptors are transduced into intracellular signals through heterotrimeric G proteins. Recently, there have been reports which show diverse coupling patterns of ligand-activated receptors to the members of Gq family alpha subunits. In order to shed some light on these complex signal processing networks, interactions between G alpha q family of G protein and neurokinin-2 receptor as well as muscarinic M1 receptor, which are considered to be new therapeutic targets in asthma, were studied. Using washed membranes from Cos-7 cells co transfected with different G alpha q and receptor cDNAs, the receptors were stimulated with various concentrations of carbachol and neurokinin A and the agonist-dependent release of [3H]inositol phosphates through phospholipase C beta 1 activation was measured. Differential coupling of G alpha q family of G-protein to muscarinic M1 receptor and neurokinin-2 receptor was observed. The neurokinin 2 receptor shows a ligand-mediated response in membranes co-transfected with G alpha q, G alpha 11 and G alpha 14 but not G alpha 16 and the ability of the muscarinic M1 receptor to activate phospholipase C through G alpha q/11 but not G alpha 14 and G alpha 16 was demonstrated. Clearly G alpha q/11 can couple M1 and neurokinin-2 receptor to activate phospholipase C. But, there are differences in the relative coupling of the G alpha 14 and G alpha 16 subunits to these receptors. PMID- 9875471 TI - Isolation and characterization of beta-galactoside specific lectin from Korean mistletoe (Viscum album var. coloratum) with lactose-BSA-sepharose 4B and changes of lectin conformation. AB - Lectins and its A- and B-chains from Korean mistletoe (Viscum album var. coloratum) were isolated by affinity chromatography on the Sepharose 4B modified by lactose-BSA conjugate synthesized by reductive amination of ligand (lactose) to epsilon-amino groups of lysine residues of spacer (BSA) after reduction by NaCNBH3. The lactose-BSA conjugate was coupled to Sepharose 4B activated by cyanogen bromide. The molecular weight determined by SDS-PAGE were a 31 kD of A chain and a 35 kD of B-chain. Amino acid analysis and N-terminal sequencing were performed. The effects of pH, temperature and guanidine chloride on the conformation of the lectin were investigated by measuring its intrinsic fluorescence and compared with its hemagglutinating activities. Blue shift was detected on the acidic pH and there was a close relationship between activities and conformation of the lectin. Under denaturing conditions, the tryptophan emission profile of lectin showed typical denaturational red shift which also correspond to the conformations and activity of lectin. PMID- 9875472 TI - PAF antagonistic activity of 2-hydroxy-3-methoxybenzoic acid glucose ester from Gentiana scabra. AB - In order to find out anti-platelet activating factor (PAF) from natural resources, Korean medicinal plants used for the treatments of peripheral circulation disorders were tested for their possible protective effects on PAF induced anaphylactic shock. From the above screening, the methanol extract of Gentiana scabra showed a potent antagonistic activity against PAF. Water suspension of the extract was partitioned with CH2Cl2 and EtOAc, successively. The EtOAc fraction which showed the highest activity was chromatographed on silica gel to yield 6 fractions. From the fraction which showed higher PAF antagonistic activity than the other fractions, compound 1 was isolated by recrystallization. On the basis of spectral data, compound 1 was identified as 2 hydroxy-3-methoxybenzoic acid glucose ester. The compound prevented the mice from the PAF-induced death at a dose of 300 micrograms/mouse. PMID- 9875473 TI - Synthesis and antifungal evaluation of 6-(N-arylamino)-7-methylthio-5,8 quinolinediones. AB - A series of 6-(N-arylamino)-7-methylthio-5,8-quinolinedione derivatives 4a-4l was newly synthesized for the evaluation of antifungal activity. 6-(N-Arylamino)-7 methylthio-5,8-quinolinediones were prepared by regioselective nucleophilic substitution of 6,7-dichloro-5,8-quinolinediones with arylamines in the presence of Ce3+, and Na2S/dimethylsulfate. The MIC values of 4a-4l were determined for antifungal susceptibility in vitro against Candida species by agar streak method. The derivatives 4a-4l had generally potent antifungal activities against all human pathogenic fungi. Especially they had the most potent activity against C. krusei at 12.5-0.8 micrograms/ml. Compounds 4d, 4g, 4h, 4j and 4k had more potent antifungal activities than fluconazole. Compounds 4g and 4h completely inhibited the fungal growth at 0.8-6.3 micrograms/ml against all Candida species, while fluconazole inhibited the growth at 25 micrograms/ml. The compounds such as 4g and 4h containing an N-(4-bromo-2-methylphenyl)- or N-(4-bromo-3 methylphenyl)amino substituent exhibited the most potent antifungal activities. PMID- 9875474 TI - Synthesis and biological activity of 5-hydroxy-4-quinolones and 5-methoxy-4 quinolones as truncated acridones. AB - A series of 5-hydroxy-4-quinolone (3) and 5-methoxy-4-quinolone (4) derivatives were synthesized as truncated acridone analogues and evaluated for antitumor, antiherpes and antituberculosis activities. Among them 5-hydroxy-8-methoxy quinolone showed potent antitumor activity (IC50 = 17.7 microM for HL60) which was greater than that of acronycine. However, these compounds didn't show any significant antiherpes or antituberculosis activity. PMID- 9875475 TI - Design and synthesis of bioisosteres of ultrapotent protein kinase C (PKC) ligand, 5-acetoxymethyl-5-hydroxymethyl-3-alkylidene tetrahydro-2-furanone. AB - Three compounds, 5-(acetoxymethyl)-5-(hydroxymethyl)-3-tetradecyl-2,5-dihydro-2 furanone (3), 5-(acetoxymethyl)-5-(hydroxymethyl)-3,3-dihexyltetrahydro-2-furano ne (4) and 5-(acetoxymethyl)-5-(hydroxymethyl)-3,3-dioctyltetrahydro-2-furano ne (5), were designed and synthesized as surrogates of the ultrapotent DAG analogue, 5-(acetoxymethyl)-5-(hydroxymethyl) 3-[(Z)-tetradecylidene]tetrahydro-2-furanone (1), a compound that showed high affinity for PKC-alpha (Ki = 35 nM) in a competition binding assay with [3H-20]phorbol-12,13-dibutyrate (PDBU). In an attempt to overcome the problem of generating geometrical E- and Z-isomers, as encountered with 1, the double bond was moved to an endocyclic location as in 3, or an additional alkyl chain was appended to C3 to give the corresponding 3,3 dialkyl saturated lactones (4 and 5). The lactone was constructed from glycidyl-4 methoxyphenyl ether in 5 steps. The target compounds showed reduced binding affinities for PKC-alpha with Ki values of 192 nM (3), 4,829 nM (4), and 2,812 nM (5), respectively. These results indicate that constrained DAG analogues having a tetrahydro-2-furanone template are effectively discriminated by PKC-alpha in terms of the direction of the long alkyl chain connected to the 3-position. PMID- 9875476 TI - Potential antitumor alpha-methylene-gamma-butyrolactone-bearing nucleic acid base. 3. Synthesis of 5'-methyl-5'-[(6-substituted-9H-purin-9-yl)methyl]-2'-oxo 3'- methylenetetrahydrofurans. AB - Search for a new alpha-methylene-gamma-butyrolactone-bearing 6-substituted purine as a potential antitumor agent has led to synthesize seven, hitherto unreported, 5'-Methyl-5'-[(6-substituted-9H-purin-9-yl)methyl]-2'-oxo-3'- methylenetetrahydrofurans (H, Cl, I, CH3, NH2, SH, > C=O) (6a-g). These include 5'-Methyl-5'-[(9H-purin-9-yl)methyl]-2'-oxo-3'-methylenetetrahydrofur ans (6a), 5'-Methyl-5'-[(chloro-9H-purin-9-yl)methyl]-2'-oxo-3'- methylenetetrahydrofurans (6b), 5'-Methyl-5'-[(6-iodo-9H-purin-9-yl) methyl]-2'-oxo-3' methylenetetrahydrofurans (6c), 5'-Methyl-5'-[(6-methyl-9H-purin-9-yl) methyl]-2' oxo-3'-methylenetetrahydrofurans (6d), 5'-Methyl-5'-[(9H-adenin-9-yl)methyl]-2' oxo-3-methylenetetrahy drofurans (6e), 5'-Methyl-5'-[(6-mercapto-9H-purin-9-yl) methyl]-2'-oxo-3'-methylenetetrahydrofurans (6f) and 5'-Methyl-5'-[(9H hypoxanthin-9-yl)methyl]-2'-oxo-3'- methylenetetrahydrofurans (6g) which were made by the Reformatsky-type reaction of ethyl alpha-(bromomethyl) acrylate with the corresponding (6-substituted-9H-purin-9-yl)-2-propanone intermediates (5a-g). These ketone intermediates 5a-g, 1-(9H-purin-9-yl)-2-propanone (5a), 1-(6-chloro 9H-purin-9-yl)-2-propanone (5b), 1-(6-iodo-9H-purin-9-yl)-2-propanone (5c), 1-(6 methyl-9H-purin-9-yl)-2-propanone (5d), 1-(9H-adenin-9-yl)-2-propanone (5e), 1-(6 mercapto-9H-purin-9-yl)-2-propanone (5f), and 1-(9H-hypoxanthin-9-yl)-2-propanone (5g) were directly obtained by the alkylation of the 6-substituted purine bases with the chloroacetone in the presence of K2CO3 (or NaH) under DMF (or DMSO). The preliminary in vitro cytotoxicity assay for the synthetic alpha-methylene-gamma butyro-lactone compounds (6a-g) were determined against three cell lines (PM-3A, P-388, and K-562) and showed the moderate antitumor activity (IC50 ranged from 1.4 to 4.3 micrograms/ml) with the compound 5'-methyl-5'-[(9H-hypoxanthin-9 yl)methyl]-2'-oxo-3'- methylenetetrahydrofuran (6g) showing the least antitumor activity. PMID- 9875477 TI - Synthesis and antitumor evaluation of cis-(1,2-diaminoethane) dichloroplatinum (II) complexes linked to 5- and 6-methyleneuracil and -uridine analogues. AB - The search for platinum (II)-based compounds with improved therapeutic properties was prompted to design and synthesize a new family of water-soluble, third generation cis-diaminedichloroplatinum (II) complexes linked to uracil and uridine. Six heretofore unreported uracil and uridine-platinum (II) complexes are; [N-(uracil-5-yl-methyl)ethane-1,2-di-amine]dichloroplatinum (II) (3a), [N (uracil-6-yl-methyl)ethane-1,2-diamine] dichloroplatinum (II) (3b), ?[N-(2', 3',5'-tri-O-acetyl)uridine-5-yl-methyl] ethane-1,2-diamine?dichloroplatinum (II) (6a), ?[N-(2',3', 5'-tri-O-acetyl) uridine-6-yl-methyl)ethane-1,2 diamine?dichloroplatinum (II) (6b), [N-(uridine-5-yl-methyl)ethane-1,2 diamine]dichloroplatinum (II) (7a), [N-(uridine-6-yl-methyl)ethane-1,2 diamine]dichloroplatinum (II) (7b). These analogues were prepared from the key starting materials, 5-chloromethyluracil (1a) and 6-chloromethyluracil (1b) which were reacted with ethylenediamine to afford the respective 5-[(2 aminoethyl)amino] methyluracil (2a) and 6-[(2-aminoethyl)amino]methyluracil (2b). The cis-platin complexes 3a and 3b were obtained through the reaction of the respective 2a and 2b with potassium tetrachloroplatinate (II). The heterocyclic nucleic acid bases 1a and 1b were efficiently introduced on the beta-D-ribose ring via a Vorbruggen-type nucleoside coupling procedure with hexamethyldisilazane, trimethylchlorosilane and stannic chloride under anhydrous acetonitrile to yield the stereospecific beta-anomeric 5-chloromethyl-2',3',5' tri-O-acetyluridine (4a) and 6-chloromethyl-2',3',5'-tri-O-acetyluridine (4b), respectively. The nucleosides 4a and 4b were coupled with ethylenediamine to provide the respective 5-[(amino-ethyl)amino]methyl-2',3',5'-tri-O-acetyluridine (5a) and 6-[(aminoethyl)amino] methyl-2',3',5'-tri-O-acetyluridine (5b). The diamino-uridines 5a and 5b were reacted with potassium tetrachloroplatinate (II) to give the novel nucleoside complexes, 6a and 6b, respectively which were deacetylated into the free nucleosides, 7a and 7b by the treatment with CH3ONa. The cytotoxic activities were evaluated against three cell lines (FM-3A, P-388 and J-82) and none of the synthesized compounds showed any significant activity. PMID- 9875478 TI - Kanamycin acetyltransferase gene from kanamycin-producing Streptomyces kanamyceticus IFO 13414. AB - A kanamycin producer, Streptomyces kanamyceticus IFO 13414 is highly resistant to kanamycin. Cloning of the kanamycin resistance genes in S. lividans 1326 with pIJ702 gave several kanamycin resistant transformants. Two transformants, S. lividans SNUS 90041 and S. lividans SNUS 91051 showed similar resistance patterns to various aminoglycoside antibiotics. Gene mapping experiments revealed that plasmids pSJ5030 and pSJ2131 isolated from the transformants have common resistant gene fragments. Subcloning of pSJ5030 gave a 1.8 Kb gene fragment which showed resistance to kanamycin. Cell free extracts of S. lividans SNUS 90041, S. lividans SNUS 91051 and subclone a S. lividans SNUS 91064 showed kanamycin acetyltransferase activity. The detailed gene map is included. PMID- 9875479 TI - Distribution of bacteria with the arylsulfate sulfotransferase activity. AB - This study is to predict the possible roles of the arylsulfate sulfotransferase (ASST) in the microorganism. At first we studied the spectrum of a distribution of the ASST enzyme through about 1,300 bacteria and the several selected strains were compared with Klebsiella K-36 previously reported in the level of DNA homology using the Southern blot method. From this study, we could predict that this enzyme would not exist in specific bacteria and it might not be a critical enzyme for the life of bacteria. PMID- 9875480 TI - Studies of structure activity relationship of flavonoids for the anti-allergic actions. AB - The structure activity relationship of flavonoids for anti-allergic actions was studied by determining the IC50 values for the degranulation. The hexosaminidase release from RBL-2H3 cells (degranulation marker) was employed as an estimate for the anti-allergic actions. Among 22 flavonoid compounds tested, luteolin, apigenin, diosmetin, fisetin, and quercetin were found to be most active with IC50 values less than 10 microM. PMID- 9875482 TI - A new epoxynaphthoquinol from Rumex japonicus. AB - A new epoxynaphthoquinol derivative, 3-acetyl-2-methyl-1, 5-dihydroxy-2,3 epoxynaphthoquinol (I), was isolated from the root of Rumex japonicus. The structure was elucidated by high field 1D and 2D NMR techniques. PMID- 9875481 TI - Yomogin, an inhibitor of nitric oxide production in LPS-activated macrophages. AB - In activated macrophages the inducible form of nitric oxide synthase (i-NOS) generates high amounts of toxic mediator, nitric oxide (NO) which contributes to the circulatory failure associated with septic shock. A sesquiterpene lactone compound (yomogin) isolated from medicinal plant Artemisia princeps Pampan inhibited the production of NO in LPS-activated RAW 264.7 cells by suppressing i NOS enzyme expression. Thus, yomogin may be a useful candidate for the development of new drugs to treat endotoxemia and inflammation accompanied by the overproduction of NO. PMID- 9875483 TI - Signal transduction in wound pharmacology. AB - Growth factors such as TGF-beta, PDGF and FGF are thought to play important roles in wound healing. However, their biological activity and signal transduction during wound repair remain poorly understood. Growth factors are often ligands for receptor tyrosine kinase and receptor serine/threonine kinases. With recent advances in signal transduction by receptor kinases, we are beginning to understand the underlying mechanism of how growth factors may regulate cutaneous wound repair. In this paper, we will describe the pharmacological effects of growth factors on wound healing, and discuss the potential underlying signaling mechanisms. Thus, we hope to provide the basis for designing more specific therapeutics for wound healing in the near future. PMID- 9875484 TI - Radiolabelled monoclonal antibodies (McAb): an alternate approach to the conventional methods for the assessment of cardiomyocyte damage in an experimental brain-death pig model. AB - The present study was carried out to determine the possible use of cTn-I in the cardiac myofibrillar architecture, as a potential target for in vivo radioimmunodetection of cardiac damage in a brain death pig model. Radioiodination of the anti-cTn-I 5F4 McAb was carried out by lactoperoxidase method. The percentage iodine incorporation achieved was 70-75%. The radioiodinated McAbs were purified on Sephadex G-25 column and characterised by Paper chromatography, Phast Gel electrophoresis and electroimmunoblotting. Radioiodinated anti-cTn-I 5F4 McAbs were employed alongside Pyrophosphate (Tc99m PPi) and Thallium201 chloride (Tl201) in 24 landrace pigs (brain-dead = 18 & sham operated = 6). The percentage cardiac uptake of the radiolabelled antibody injected dose was significantly higher in the brain dead animals (0.196%) as compared to that of sham-operated animals (0.11%). Specific in vivo localization of radiolabelled McAbs in the infarcted cardiac tissue was confirmed by computer aided reconstruction of 3-D images of the isolated heart. The preliminary results of the study revealed preferential uptake of radiolabelled antibody at the site of myocyte damage resulting from artificially induced brain death. PMID- 9875485 TI - Percutaneous absorption and model membrane variations of melatonin in aqueous based propylene glycol and 2-hydroxypropyl-beta-cyclodextrin vehicles. AB - Percutaneous absorption and model membrane variations of melatonin (MT) in aqueous-based propylene glycol and 2-hydroxypropyl-beta-cyclodextrin vehicles were investigated. The excised hairless mouse skin (HMS) and two synthetic ethylene vinyl acetate (EVA) and microporous polyethylene (MPE) were selected as a model membrane. The solubility of MT was determined by phase equilibrium study. The vertical Franz type cell was used for diffusion study. The concentration of MT was determined using reverse phase HPLC system. The MT solubility was the highest in a mixture of PG and 2-HP beta CD. The percutaneous absorption of MT through excised HMS increased as the solubility increased. However, the permeability coefficient decreased and then slightly increased in a mixture of PG and 2-HP beta CD. On the other hand, both flux and permeability coefficient through EVA membrane decreased as the solubility increased. No MT was detected over 12 h after starting diffusion through MPE membrane. The flux of MT was dependent on the type of membrane selected. Flux of MT was greatest in excised HMS followed by EVA and MPE membrane. Flux of MT through EVA membrane was 5-20 times lower when compared to excised HMS. Interestingly, volumes of donor phase when MPE membrane was used, significantly increased during the study period. The HMS might be applicable to expect plasma concentration of MT in human subjects based on flux and pharmacokinetic parameters as studied previously. The current studies may be applied to deliver MT transdermally using aqueous-based vehicles and to fabricate MT dosage forms. PMID- 9875486 TI - Studies on protective effect of DA-9601, Artemisia asiatica extract, on acetaminophen- and CCl4-induced liver damage in rats. AB - The hepatoprotective effect of DA-9601, a quality-controlled extract of Artemisia asiatica, on liver damage induced by acetaminophen (APAP) and carbon tetrachloride (CCl4) was investigated by means of serum-biochemical, hepatic biochemical, and histopathological examinations. Doses of DA-9601 (10, 30, or 100 mg/kg) were administered intragastrically to each rat on three consecutive days i.e. 48 h, 24 h and 2 h before a single administration of APAP (640 mg/kg, i.p.) or CCl4 (2 ml/kg, p.o.). Four h and 24 h after hepatotoxin treatment, the animals were sacrificed for evaluation of liver damage. Pretreatment of DA-9601 reduced the elevation of serum ALT, AST, LDH and histopathological changes such as centrilobular necrosis, vacuolar degeneration and inflammatory cell infiltration dose-dependently. DA-9601 also prevented APAP- and CCl4-induced hepatic glutathione (GSH) depletion and CCl4-induced increase of hepatic malondialdehyde (MDA), a parameter of lipid peroxidation, in a dose-dependent manner. These findings suggest that pretreatment with DA-9601 may reduce chemically induced liver injury by complex mechanisms which involve prevention of lipid peroxidation and preservation of hepatic GSH. PMID- 9875487 TI - The development of a new method to detect the adulteration of commercial aloe gel powders. AB - Simple and accurate methods to detect the adulteration of commercial aloe gel powder were developed. Crude polysaccharide in aloe gel powder was isolated by precipitating with excess ethyl alcohol and total hexose in isolated polysaccharide was determined by Dubois assay. After hydrolysis of non-dialysable polysaccharides, resultant free sugar was determined by gas chromatography for sugar recognition and ash contents was considered simultaneously. In some products, the content of ash was very low while the content of total hexose was very high. And polysaccharides of these products revealed typical dextran pattern, therefore, these products could be identified that adulterated with commercial maltodextrin. The content of maltodextrin in adulterated product was determined by HPLC and TLC analysis which could be adopted as a part of a certification process. PMID- 9875488 TI - Physicochemical properties of recombinant hepatitis B surface antigen expressed in mammalian cell (C127). AB - The physicochemical properties of recombinant hepatitis B surface antigen (r HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCl density gradient method was 1.19 g/ml and the isoelectric point by Mono P HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. Fluorophore-assisted-carbohydrate electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and its major component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derived r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B. PMID- 9875489 TI - Comparative activities of novel beta-lactamase inhibitors, 6-exomethylene penamsulfones (CH1240, CH2140) in experimental mouse infection model. AB - The antibacterial activity of novel beta-lactamase inhibitors, 6-exomethylene penamsulfones (CH1240, CH2140), has been compared in vivo with that of sulbactam and clavulanic acid against beta-lactamase producing strains. In vivo microbiological assessment was used as experimental mouse infection model by gram negative strains. Against Pseudomonas aeruginosa F0013, cefoperazone/CH1240 was slightly less active than sulbactam. Ampicillin/CH1240 was more active than sulbactam against Citrobacter diversus species. That of ampicillin/CH2140 was less effective than sulbactam against Escheriachia coli 3457. Especially against Citrobacter diversus 2046E, amoxicillin/CH2140 was the most potent and amoxicillin/CH1240 was slightly more active than clavulanic acid. Consequently the difference in efficacy between the drug combinations appears to be related to the degree of protection afforded the animals by the beta-lactamase inhibitors. CH1240 and CH2140 are promising new agents and should undergo further investigations. PMID- 9875490 TI - Protective effect of taurine on TNBS-induced inflammatory bowel disease in rats. AB - We had previously reported that the protective effect of taurine against indomethacin-induced gastric mucosal injury was due to its antioxidant effects, which inhibited lipid peroxidation and neutrophil activation. In this study, we examined the effect of taurine on reducing the inflammatory parameters of trinitrobenzene sulfonic acid (TNBS)-induced inflammatory bowel disease (IBD) in rats. In order to induce IBD, ethanolic TNBS was given to rats intracolonically. Then they received 500 mg/kg/day of taurine orally and were sacrificed one week after IBD induction. While ulceration and inflammation of distal colon with formation of granuloma in the vehicle-treated IBD rats two days after administration of TNBS were observed, treatment with taurine ameliorated colonic damage and decreased the incidence of diarrhea and adhesion. Also, colon weight as an index of tissue edema, which was markedly increased in the IBD rats, became significantly lower after taurine treatment. Myeloperoxidase (MPO) activity in the vehicle-treated IBD rats was substantially increased, compared with that of normal control. The taurine-treated animals significantly reduced MPO activity (35% lower) when compared with that of the vehicle-treated animals. Taurine treatment decreased both basal and formyl-methionyl leucyl phenylalanine stimulated reactive oxygen generation from colonic tissue in the IBD rats. These results suggest that the administration of taurine reduce the inflammatory parameters in this IBD rat model by increasing defending capacity against oxidative damage. PMID- 9875491 TI - Vaccination with an ovalbumin/interleukin-4 fusion DNA efficiently induces Th2 cell-mediated immune responses in an ovalbumin-specific manner. AB - To more effectively drive immune responses toward antigen-specific T helper type 2 (Th2) cell-mediated responses, we constructed a mammalian expression vector (pOVA/IL4) carrying a fused gene in which the ovalbumin (OVA) cDNA was covalently linked to murine interleukin-4 (IL-4) cDNA. A biologically active OVA/IL4 protein was expressed by the transfected COS cells with the pOVA/IL4 DNA, as demonstrated by Western blotting and cytokine bioassay. Intramuscular injection of BALB/c mice with the pOVA/IL4 DNA increased both the production of OVA-specific IL-4 by CD4+ T cells and the ratio of anti-OVA IgG1 to anti-OVA IgG2a isotypes, while the injection with the pOVA DNA alone, or with the mixture of the pOVA and pIL4 DNA did no or little increase. Furthermore, the OVA-specific, Th2 cell-mediated immune responses were significantly enhanced by multiple injections with the pOVA/IL4 DNA. These studies indicate that the direct linkage of an OVA gene to an IL-4 gene in the expression plasmid confines the effects of IL-4 to the OVA specific cells, efficiently driving the immune response toward OVA-specific, Th2 cell-mediated responses. PMID- 9875492 TI - Immunological properties of recombinant hepatitis B surface antigen expressed in mammalian cell (C127). AB - We examined the immunological properties of the recombinant hepatitis B surface antigen (r-HBsAg) which was expressed in mammalian cell (C127). The cross immunity of r-HBsAg and plasma-derived hepatitis B surface antigen (p-HBsAg) were tested using Western blotting and ELISA with guinea pig polyclonal antibody and naturally infected human-derived antibody and the both antigens show the same results in their response pattern and intensity, which indicate they have a good cross-immunity. From the measurement of ED50 after formalin- or heat inactivation, both r-HBsAg and p-HBsAg showed ED50 of 0.2-0.3 in formalin inactivation, while r-HBsAg was 0.05-0.09 and p-HBsAg was 0.03-0.07 in heat inactivation, which means heat-inactivation method is 3-4 times superior in immunogenicity. In the immunopersistency test performed in guinea pig for the period of 3 months with two different adjuvants, antibody titer was 34.2 with muramyl dipeptide adjuvant, which was 1.8 times greater than the antibody titer of 18.9 with AlPO4 adjuvant. The mutagenicity of r-HBsAg, examined by Ames test and micronucleus test, were all negative. In conclusion, r-HBsAg has the same cross-immunity with p-HBsAg, and heat-inactivation method and muramyl dipeptide adjuvant allow development of r-HBsAg vaccine with excellent immunogenicity. PMID- 9875493 TI - Changes of tissue factor activity on inflammatory stimulus and aging in rat. AB - Tissue factor (TF), a principal initiator of the vertebrate coagulation cascade, is expressed in organ tissues, cells and blood. TF is known to be induced in endothelial cells, monocytes and macrophages by inflammatory stimuli and in many pathologic conditions. By using the modified method for in vivo TF activity assay, we found that turpentine oil injection as an inflammatory stimulus also induced the TF activity in lung and brain tissues of rats. And the age-related increase in TF activity was observed in healthy rat brain tissue. PMID- 9875494 TI - Isolation and identification of chondroitin sulfates from the mud snail. AB - Chondroitin sulfates were isolated from the mud snail. For the quantitative analysis of enzymatic digestion products of isolated chondroitin sulfates, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitinase ABC, three unsaturated disaccharides 2-acetamide 2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-D-galactose (delta Di-OS), 2 acetamide-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D galactose (delta Di-6S) and 2-acetamide-2-deoxy-3-O-(beta-D-gluco-4 enepyranosyluronic acid)-4-O-sulfo-D-galactose (delta Di-4S) were produced from the mud snail chondroitin sulfates. The analysis showed that relative proportion of delta Di-OS/delta Di-6S/delta Di-4S was 58.7/3.1/38.2. The immunomodulating activity of chondroitin sulfate was examined by cell proliferation assay and these results suggest that it might be a immunosuppressant. PMID- 9875495 TI - Synthesis and pharmacokinetic profile of 3-methoxymethyl cephalosporin prodrugs. AB - Preparation and biological activity of prodrug-type 3-methoxymethyl cephalosporins were described. From the mixtures, R- and S-prodrugs were separated and their absolute configurations were determined, and also their bioavailability was investigated. PMID- 9875497 TI - Formation, properties and antimicrobial activities of cotton xanthate-Cu(II) homosulfamine complex. AB - To develop a cotton derivatives with prolonged antimicrobial activities, homosulfamine (Hs) was coupled to cotton xanthate (CX) via chelate bond in the presence of Cu(II) ion by one- and two-bath processes. In one-bath process, CX was treated with Cu(II)-Hs solution. In two-bath process, CX was treated with Cu(II) ion solution to produce CX-Cu(II) complex, which was isolated and treated in turn with Hs solution. Effects of concentration, Cu(II)/Hs ratio, and pH on the binding of Hs were investigated at 10 degrees C. In one-bath process, binding of Hs took place readily with optimum pH around 5-6. The amount of binding increased to give a maximum within 5 min and decreased slowly to establish an equilibrium within an hour. In two-bath process, binding of Hs was much lower than that of one-bath process. Release of Hs from CX-Cu(II)-Hs was investigated by batch and flow method. Antimicrobial activities of CX-Cu(II)-Hs were tested against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli and it showed prolonged activity compared to that of free Hs. PMID- 9875496 TI - Enantiomeric ratio of shikonin derivatives as a possible key for the determination of the origin of lithospermi radix. AB - An HPLC method was developed to resolve the enantiomers of shikonin derivatives of the Lithospermi Radix. The optimum mobile phase on a Chiracel AD column was 5% isopropanol in n-hexane with flow rate of 1 ml/min. Establishment of this method made possible to determine the ratios of shikonin/acetylshikonin or alkanin/acetylalkanin in the same root. The correlation of the ratios of these substance pairs appeared characteristic for the country where they were originated from. All of the Korean species showed significantly higher ratios of shikonin/acetylshikonin and alkanin/acetylalkanin than the Chinese ones. This method would be useful to determine the origin of Lithospermi Radix. PMID- 9875498 TI - Degradation of acharan sulfate and heparin by Bacteroides stercoris HJ-15, a human intestinal bacterium. AB - When glycosaminoglycan (GAG)-degrading enzymes were measured in normal human stool suspensions, all 5 tested different stools degraded titrable heparin and acharan sulfate. GAG-degrading bacteria were screened from the isolates of human stools. Among them, HJ-15 had the most potent activities of heparinases (GAGs degrading enzymes). However, HJ-15 produced the enzyme even if in the media without heparin. Acharan sulfate lyase was induced by acharan sulfate and heparin. Heparinase production was also induced by these GAGs. These enzymes, acharan sulfate lyase and heparinase, were produced in exponential and stationary phase of HJ-15 growth, respectively. Optimal pHs of the acharan sulfate lyase and heparinase activities were 7.2 and 7.5, respectively. The biochemical properties of HJ-15 was similar to those of B. stercoris. However, difference from B. stercoris was utilization of raffinose. This HJ-15 also degraded chondroitin sulfates A and C. PMID- 9875499 TI - Antitumor activity of 7-[2-(N-isopropylamino)ethyl]-(20S)-camptothecin, CKD602, as a potent DNA topoisomerase I inhibitor. AB - We developed a novel water-soluble camptothecin analogue, CKD602, and evaluated the inhibition of topoisomerase I and the antitumor activities against mammalian tumor cells and human tumor xenografts. CKD602 was a nanomolar inhibitor of the topoisomerase I enzyme in the cleavable complex assay. CKD602 was found to be 3 times and slightly more potent than topotecan and camptothecin as inhibitors of topoisomerase, respectively. In tumor cell cytotoxicity, CKD602 was more potent than topotecan in 14 out of 26 human cancer cell lines tested, while it was comparable to camptothecin. CKD602 was tested for the in vivo antitumor activity against the human tumor xenograft models. CKD602 was able to induce regression of established HT-29, WIDR and CX-1 colon tumors, LX-1 lung tumor, MX-1 breast tumor and SKOV-3 ovarian tumor as much as 80, 94, 76, 67, 87% and 88%, respectively, with comparable body weight changes to those of topotecan. Also the therapeutic margin (R/Emax: maximum tolerance dose/ED58) of CKD602 was significantly higher than that of topotecan by 4 times. Efficacy was determined at the maximal tolerated dose levels using schedule dependent i.p. administration in mice bearing L1210 leukemia. On a Q4dx4 (every 4 day for 4 doses) schedule, the maximum tolerated dose (MTD) was 25 mg/kg per administration, which caused great weight loss and lethality in < 5% tumor bearing mouse. This schedule brought significant increase in life span (ILS), 212%, with 33% of long-term survivals. The ex vivo antitumor activity of CKD602 was compared with that of topotecan and the mean antitumor index (ATI) values recorded for CKD602 were significantly higher than that noted for topotecan. From these results, CKD602 warrants further clinical investigations as a potent inhibitor of topoisomerase I. PMID- 9875500 TI - Structural analogues of cumambrin B from the flower of Chrysanthemum boreale. AB - The structural analogues of cumambrin B (1, 2, 3, 4) were isolated from the flower of Chrysanthemum boreale Makino. The structures of compounds were determined by two-dimensional 1H-1H COSY and 13C-1H COSY spectra with the aid of homonuclear and heteronuclear double resonance experiment. The stereochemistry of compounds has been verified from single crystal X-ray diffraction of cumambrin A (2). The antimicrobial activities of these guaianolides have been studied. PMID- 9875501 TI - Naphthazarin derivatives: synthesis, cytotoxic mechanism and evaluation of antitumor activity. AB - The rate of the GSH conjugate formation, the inhibition of DNA topoisomerase-I and the cytotoxic activity against L1210 cells of the naphthoquinones showed the same order; 5,8-dimethoxy-1,4-naphthoquinone (DMNQ) > 6-(1-hydroxyethyl)-DMNQ > 2 (1-hydroxyethyl)-DMNQ; the steric hindrance of the substituents, particularly 2 substutuent, in reacting with cellular nucleophiles must be the main cause for lowering the bioactivities. Acetylation of 2-(1-hydroxyethyl)-DMNQ producing 2 (acetyloxyethyl)-DMNQ potentiated the bioactivities; 2-(1-hydroxyethyl)-DMNQ did not react with GSH and the enzyme, and showed ED50 of 0.680 microgram/ml, whereas the values of 2-(1-acetyloxyethyl)-DMNQ were the conjugate formation of 0.14 microM, IC50 value of 81 microM for the enzyme inhibition and ED50 of 0.146 microgram/ml for the cytotoxcity. Furthermore, the acetylation 2-(1-hydroxyethyl) DMNQ (T/C, 119%) enhanced the T/C values for the mice bearing S-180 tumor [T/C of 2-(1-acetyloxyethyl)-DMNQ, 276%]. It was assumed that the difference in bioactivities ensued by acetylation was based on the mechanism of the so-called bioreductive alkylation. PMID- 9875502 TI - Synthesis of 6-aziridinylbenzimidazole derivatives and their in vitro antitumor activities. AB - In search for new antitumor agents, twelve 6-aziridinylbenzimidazole derivatives were synthesized and their cytotoxicities were tested against three cancer cell lines (mouse lymphocytic leukemia P388 and B16, and human gastric carcinoma SNU 16). From 4-amino-3-nitrotoluene as the starting material, 2 (acetoxymethyl)benzimidazoles (5a-d) were obtained by Phillips reaction. These benzimidazoles were then reacted with Fremy's salt to give a mixture of three 2 (acetoxymethyl) (8a-c) and four 2-(hydroxymethyl)benzimidazole-4,7-diones (9a-d). Addition of these quinones with aziridine afforded 6-aziridinyl-2-(acetoxymethyl) (10a-c) and 6-aziridinyl-2-(hydroxymethyl)benzimidazole-4,7-diones (11a-d). Utilizing 2-(hydroxymethyl)benzimidazole-4,7-diones (9b,d), esters 10d and 13e-h were prepared by the sequential reactions of esterification and addition. The synthesized compounds show potent cytotoxicity against all of three cell lines tested. The cytotoxicities of 10a-d or 11a-d against SNU-16 were superior to those of 13e-h, and were equal to or slightly higher than that of mitomycin C. Compounds 11a-d were slightly more cytotoxic than 10a-d in all cell lines tested. PMID- 9875505 TI - Thymol derivatives from Carpesium divaricatum. AB - Four thymol derivatives, 2,5-dimethoxythymol (1), 2-methoxythymol isobutyrate (2), 10-isobutyloxy-8,9-epoxythymolisobutyrate (3) and 10-(2-methylbutyloxy)-8,9 epoxythymolisobutyrate (4) were isolated from the aerial parts of Carpesium divaricatum. The structures were elucidated by high field 1D and 2D NMR techniques. PMID- 9875503 TI - Immunomodulating activity of DW-116, a new quinolone antibiotic. AB - DW-116, (1-(5-fluoro-2-pyridyl)-6-fluoro-7-(4-methyl-1-piperazinyl)-1, 4-dihydro 4-oxoquinoline-3-carboxylic acid hydrochloride), is a new quinolone antibiotic with a broad antibacterial spectrum against G(+) and G(-) bacteria. DW-116 was evaluated for the immunomodulating activities, which is one of the efforts to investigate the mechanism of action related to the good in vivo antibacterial efficacy. The results of in vitro studies revealed there was no statistically significant increase in B and T lymphocyte proliferation. But the results of in vivo studies showed that the number of plaque forming cells (PFC), the amount of polyclonal antibodies and delayed-type hypersensitivity (DTH) were significantly increased after the repeat administration with 12 and 60 mg/kg of DW-116. Taken together, these results proposed that immunostimulating effect of DW-116 could be one of the action mechanisms for demonstrating in vivo antibacterial activities under these experimental conditions. PMID- 9875504 TI - Ginsenoside Rf2, a new dammarane glycoside from Korean red ginseng (Panax ginseng). AB - A new dammarane glycoside named ginsenoside Rf2 has been isolated from Korean red ginseng (Panax ginseng) and its chemical structure has been elucidated as 6-O [alpha-L-rhamnopyranosyl (1-->2) beta-D-glucopyranosyl]dammarane-3 beta, 6 alpha, 12 beta, 20(R), 25-pentol by chemical and spectral methods. PMID- 9875506 TI - Therapeutic effect of rebamipide on ammonia-induced gastric mucosal hemorrhagic lesion in rats. AB - Rebamipide, 2-(4-chlorobenzoylamino)-3-[2(1H)-quinolinone-4-yl]-propionic acid, a novel antipeptic ulcer agent, has been reported to prevent various acute experimental gastric mucosal lesions and to accelerate the healing of chronic ulcers. Therapeutic effect of rebamipide was investigated with regard to the inhibitory effect on xanthine oxidase activity and type conversion of the enzyme which play a profound role in oxygen radicals generation system. Intraperitoneal administration of rebamipide at 60 mg/kg body weight reduced the xanthine oxidase activity, lipid peroxide content in ammonia induced hemorrhagic lesion. These results suggest that the therapeutic effect of rebamipide on gastric mucosal lesion may be in part due to the inhibitory activity of xanthine oxidase and type conversion rate of the enzyme. PMID- 9875507 TI - Inhibitory action of YJA20379, a new proton pump inhibitor on Helicobacter pylori growth and urease. AB - The activities of two types of antiulcer agents against 9 strains of Helicobacter pylori (H. pylori) were determined by the agar dilution method. The antiulcer agents were YJA20379, a newly synthesized proton pump inhibitor developed by Yung Jin Pharmaceutical company, and omeprazole. Both compounds were found to have significant activities against this organism. The MIC values of YJA20379 and omeprazole were 11.7 and 31.25 micrograms/ml, respectively. In addition, the inhibitory potency of both compounds was investigated on H. pylori urease which is believed to be an important colonization and virulence factor in the pathogenesis of gastritis and peptic ulcers. These compounds dose-dependently inhibited urease extracted with distilled water and their IC50 values were 16.4 x 10(-5) M and 14.3 x 10(-5) M, respectively. In addition, a pH-dependent study to determine whether inhibitory potency would be activated by acid condition was performed. It was found that unlike omeprazole, YJA20379 was not affected by acid condition. To determine the inhibition pattern and optimal concentration of substrate, kinetics were evaluated at various pH levels (pH 5.0, 7.0, and 8.5). The data show that YJA20379 noncompetitively inhibited H. pylori urease and KM/Ki values were 0.96 mM/60 microM (pH 5.0), 0.56 mM/141.5 microM (pH 7.0), and 1.94 mM/34 microM (pH 8.5), respectively. Based on data obtained, it is concluded that YJA20379 is a significant inhibitor of H. pylori growth and urease and therefore, taking these results into consideration, YJA20379 might be a beneficial therapy for gastritis and peptic ulcers induced by H. pylori. PMID- 9875508 TI - Inhibitory effect of lignans from the rhizomes of Coptis japonica var. dissecta on tumor necrosis factor-alpha production in lipopolysaccharide-stimulated RAW264.7 cells. AB - The inhibitory effect of 10 lignan constituents isolated from the rhizomes of Coptis japonica var. dissecta on tumor necrosis factor (TNF)-alpha production in lipopolysaccharide (LPS)-stimulated macrophage cell line (RAW264.7 cells) has been studied. Among them, pinoresinol, woorenoside-V and lariciresinol glycoside showed significant inhibitory activities in the range from 37% to 55% at the concentration of 25 micrograms/ml. The results are first report that the lignans isolated from Coptis japonica inhibit TNF-alpha production, and suggest that the lignan components may partly participate in antiinflammatory and antiallergic effect of Coptis japonica through the inhibition of TNF-alpha production. PMID- 9875509 TI - Intestinal bacterial metabolism of flavonoids and its relation to some biological activities. AB - Flavonoid glycosides were metabolized to phenolic acids via aglycones by human intestinal microflora producing alpha-rhamnosidase, exo-beta-glucosidase, endo beta-glucosidase and/or beta-glucuronidase. Rutin, hesperidin, naringin and poncirin were transformed to their aglycones by the bacteria producing alpha rhamnosidase and beta-glucosidase or endo-beta-glucosidase, and baicalin, puerarin and daidzin were transformed to their aglycones by the bacteria producing beta-glucuronidase, C-glycosidase and beta-glycosidase, respectively. Anti-platelet activity and cytotoxicity of the metabolites of flavonoid glycosides by human intestinal bacteria were more effective than those of the parental compounds. 3,4-Dihydroxyphenylacetic acid and 4-hydroxyl-phenylacetic acid were more effective than rutin and quercetin on anti-platelet aggregation activity. 2,4,6-Trihydroxybenzaldehyde, quercetin and ponciretin were more effective than rutin and ponciretin on the cytotoxicity for tumor cell lines. We insist that these flavonoid glycosides should be natural prodrugs. PMID- 9875510 TI - A potent anti-diabetic agent from Kalopanax pictus. AB - To search for the anti-diabetic principle from the stem bark of Kalopanax pictus, seven kinds of chemical constituents including hedearagenin glycosides and phenolic glycosides were isolated. The anti-diabetic evaluation of these isolates in the streptozotocin-induced diabetic rats exhibited that kalopanaxsaponin A has a potent anti-diabetic activity in contrast to a mild activity of hedearagenin. In addition, significant hypocholesterolemic and hypolipidemic activities of kalopanaxsaponin A and hedearagenin were observed. The structure-activity relationship of kalopanaxsaponin A was also investigated in the present work. PMID- 9875511 TI - Studies on the pharmacological action of cactus: identification of its anti inflammatory effect. AB - The ethanol extracts of Opuntia ficus-indica fructus (EEOF) and Opuntia ficus indica stem (EEOS) were prepared and used to evaluate the pharmacological effects of cactus. Both the extracts inhibited the writhing syndrome induced by acetic acid, indicating that they contains analgesic effect. The oral administrations of EEOF and EEOS suppressed carrageenan-induced rat paw edema and also showed potent inhibition in the leukocyte migration of CMC-pouch model in rats. Moreover, the extracts suppressed the release of beta-glucuronidase, a lysosomal enzyme in rat neutrophils. It was also noted that the extracts showed the protective effect on gastric mucosal layers. From the results it is suggested that the cactus extracts contain anti-inflammatory action having protective effect against gastric lesions. PMID- 9875512 TI - Isolation of human CYP4F2 genomic DNA and its 5' end regulatory region structure. AB - Human cytochrome P450 4F2 shows high regioselectivity in omega-hydroxylation of stearic acid and leukotriene B4. As a first step of its regulation study, human cytochrome P450 4F2 genomic DNA was isolated from liver of a person who was administered clofibrate for 10 years. From Southern hybridization, restriction enzyme digestion and sequencing experiments, isolated genomic DNA fragment was found to contain around 32 Kb DNA and more than 20 Kb of 5' end regulatory region. Sequences of the structural gene region revealed exon 1 and exon 2. Further regulation studies would elucidate the feedback mechanisms of the oxidative degradation of fatty acids, inflammatory response and the clearance of leukotriene B4 in the liver. Furthermore, regulation study of this gene could explain the species difference in response to peroxisome proliferator and help in the safety evaluation of peroxisome proliferating chemicals to human being. PMID- 9875513 TI - Induction of differentiation of the human histocytic lymphoma cell line U-937 by hypericin. AB - Hypericin, a photosensitizing plant pigment, was found to be a potent inducer of differentiation of human myeloid leukemia U-937 cells. At a concentration of 0.2 microM, hypericin exhibited 50% growth inhibition. An effect on cell differentiation by hypericin was assessed by its ability to induce phagocytosis of latex particles, and to reduce nitroblue tetrazolium (NBT). Approximately 51% of 0.2 microM hypericin-treated cells were stained with NBT and 63% showed phagocytic activity. In order to establish whether hypericin induces differentiation of U-937 cells to macrophage or granulocyte, esterase activities and cell sizes were measured. When U-937 cells were treated with 0.2 microM and 0.15 microM of hypericin, the alpha-naphthyl acetate esterase activity was increased by 38.4% and 48.1%, respectively, but naphthol AS-D chloroacetate esterase activity was not influenced. The size of hypericin-treated cells in terms of cell mass was larger than that observed in untreated cells as determined by flow cytometry. Protein kinase C (PKC) inhibitor, NA-382, decreased the NBT reducing activity of hypericin, whereas a cAMP-dependent protein kinase A (PKA) inhibitor, H-89, did not show any influence on the differentiation. These results indicate that hypericin triggers differentiation toward monocyte/macrophage lineage by PKC stimulation. PMID- 9875514 TI - Thermotolerance inhibits various stress-induced apoptosis in NIH3T3 cells. AB - When NIH3T3 cells were exposed to mild heat and recovered at 37 degrees C for various time intervals, they were thermotolerant and resistant to subsequent stresses including heat, oxidative stresses, and antitumor drug methotrexate which are apoptotic inducers. The induction kinetics of apoptosis by stresses were determined by DNA fragmentation and protein synthesis using [35S]methionine pulse labeling. We investigated the hypothesis that thermotolerant cells were resistant to apoptotic cell death compared to control cells when both cells were exposed to various stresses inducing apoptosis. The cellular changes in thermotolerant cells were examined to determine which components are involved in this resistance. At first, the degree of resistance correlates with the extent of heat shock protein synthesis which were varied depending on the heating times at 45 degrees C and recovery times at 37 degrees C after heat shock. Secondly, membrane permeability change was observed in thermotolerant cells. When cells prelabeled with [3H]thymidine were exposed to various amounts of heat and recovered at 37 degrees C for 1/2 to 24 h, the permeability of cytosolic [3H]thymidine in thermotolerant cells was 4 fold higher than that in control cells. Thirdly, the protein synthesis rates in thermotolerant and control cells were measured after exposing the cells to the same extent of stress. It turned out that thermotolerant cells were less damaged to same amount of stress than control cells, although the recovery rates are very similar to each other. These results demonstrate that an increase of heat shock proteins and membrane changes in thermotolerant cells may protect the cells from the stresses and increase the resistance to apoptotic cell death, even though the exact mechanism should be further studied. PMID- 9875515 TI - Inhibitory effects of Bifidobacterium spp. isolated from a healthy Korean on harmful enzymes of human intestinal microflora. AB - Five hundreds of bifidobacteria were isolated from a healthy Korean and the inhibitory effects of these isolated bacteria on harmful enzymes of human intestinal microflora were examined by cocultivation of the isolated bifidobacteria with E. coli or total human intestinal microflora. In comparison with the results of E. coli or intestinal microflora cultivation, Bifidobacterium breve K-110, B. breve K-111 and B. infantis K-525 effectively inhibited harmful enzymes (beta-glucuronidase and tryptophanase) of E. coli and lowered the pH of the culture media. Also they inhibited the harmful enzymes (beta-glucosidase, beta-glucuronidase, tryptophanase and urease) and ammonia production of intestinal microflora, and lowered pH of the culture media by increasing lactic acid bacteria of intestinal microflora. When these isolated bifidobacteria were administered on mice, fecal harmful enzymes were also inhibited. Among tested bifidobacteria, B. breve K-110 had the highest inhibitory effect of fecal harmful enzymes. PMID- 9875517 TI - Isolation and identification of inhibitory compounds on TNF-alpha production from Magnolia fargesii. AB - Three TNF alpha-inhibitory lignans were isolated from the flower buds of Magnolia fargesii through bioassay-guided isolation. They were identified as eudesmin, magnolin and lirioresinol-B dimethylether on the basis of their spectroscopic data. All three lignans showed inhibitory effects on TNF-alpha production in LPS stimulated murine macrophage cell line, RAW264.7 and eudesmin showed the strongest activity (IC50 = 51 microM). PMID- 9875516 TI - Screening and isolation of antibiotic resistance inhibitors from herb materials resistance inhibition of volatile components of Korean aromatic herbs. AB - The resistance inhibitory activities of 54 odorant mixtures (essential oil) from 41 Korean aromatic herbs were tested against multi-drug resistant Staphylococcus aureus SA2, which has resistances to 10 usual antibiotics including chloramphenicol. As results, combinations of 28 kinds of samples from 21 herbs and chloramphenicol have resistance inhibitory activities in dose dependent manner. PMID- 9875518 TI - Total synthesis of fentanyl. AB - Fentanyl of a potent anilidopiperidine analgesic has been synthesized from a simple phenylethylamine by four step sequence. The key part of this synthesis involves an efficient construction of phenylethylpiperidone skeleton via aminomethano desilyltion-cyclization followed by Swern oxidation. PMID- 9875519 TI - Synthesis and in vitro evaluation of 4-substituted-1-azaanthraquinones. PMID- 9875520 TI - Resistance to macrolide-lincosamide-streptogramin B antibiotics is induced by 16 membered-ring macrolide antibiotics in Enterococcus faecalis 373. PMID- 9875521 TI - Simultaneous determination of diphenhydramine, its N-oxide metabolite and their deuterium-labeled analogues in ovine plasma and urine using liquid chromatography/electrospray tandem mass spectrometry. AB - Our studies on drug disposition in chronically instrumented pregnant sheep involve simultaneous administration of the antihistamine diphenhydramine (DPHM), its deuterated analogue ([2H10]DPHM) and their metabolites to the mother or the fetus via various routes. Such studies require sensitive and selective mass spectrometric methods for quantitation of these labeled and unlabeled compounds in order to assess comparative maternal and fetal drug metabolism. The objective of this study was to develop and validate a liquid chromatographic/tandem mass spectrometric (LC/MS/MS) method for the simultaneous quantitation of DPHM, its N oxide metabolite and their deuterium-labeled analogues in ovine plasma and urine. Samples spiked with the analytes and the internal standard, orphenadrine, were processed using liquid-liquid extraction. The extract was chromatographed on a propylamino LC column and MS/MS detection was performed in the positive ion electrospray mode using multiple reaction monitoring. The linear concentration ranges of the calibration curves for the N-oxides and the parent amines were 0.4 100.0 and 0.2-250.0 ng ml-1, respectively. In validation tests, the assay exhibited acceptable variability (< or = 15% at analyte concentrations below 2.0 ng ml-1 and < 10% at all other concentrations) and bias (< 15% at all concentrations), and the analytes were stable under a variety of sample handling conditions. Using this method, the labeled and unlabeled N-oxide metabolite was identified in fetal plasma after DPHM and [2H10]DPHM administration. This method will be used further to examine the comparative metabolism of diphenhydramine to its N-oxide metabolite in the mother and the fetus. PMID- 9875522 TI - Structures of biologically active muramyl peptides from peptidoglycan of Streptococcus sanguis. AB - The structures of major muramyl peptides derived from peptidoglycan of the oral pathogen Streptococcus sanguis were determined and the biological activity of the peptides was tested in vitro on human monocytes. The muramyl peptides, produced by muramidase digestion of the purified peptidoglycan, were separated by reversed phase high-performance liquid chromatography, either in their native form or after reduction with sodium borohydride. Chemical structures of the peptides were elucidated by a combination of matrix-assisted laser desorption/ionization time of-flight mass spectrometry, amino acid analysis, post-source decay analysis and Edman sequencing. The study revealed two distinct monomers: N-acetylglucosaminyl N-acetylmuramyl-Ala-iGln-Lys(Ala-Ala) (1), where the Ala-Ala is connected to the epsilon-amino group of lysine, and N-acetylglucosaminyl-N-acetylmuramyl-Ala-iGln Lys(Ala-Ala)-Ala-Ala (2), where an additional dialanyl residue is attached to the lysine alpha-carboxyl group. Two sets of higher oligomers (di-, tri- and tetramers), related structurally to monomers 1 or 2 were also detected. In these oligomers, the monomeric subunits are linked together by Ala-Ala-Ala bridges. The native muramyl peptides primed human monocytes in vitro for the increased production of the microbicidal superoxide radical. PMID- 9875523 TI - Molecular species of sphingomyelin: determination by high-performance liquid chromatography/mass spectrometry with electrospray and high-performance liquid chromatography/tandem mass spectrometry with atmospheric pressure chemical ionization. AB - In a sphingomyelin-enriched sample of polar lipids from bovine milk, molecular species of intact sphingomyelin were separated by normal-phase high-performance liquid chromatography and detected by mass spectrometry (MS) for structural information. First, by using electrospray with positive ionization (ESI), protonated molecules ([M + H]+) were detected. Second, in atmospheric pressure chemical ionisation (APCI+), in-source fragmentation of sphingomyelin ions led to the formation of ceramide ions. With the ceramide ions as precursors, ions representative of both the long-chain base (LCB) parts and the fatty acid (FA) parts were detected in APCI-MS/MS via collision-induced decomposition (CID). Using this procedure, it was possible to determine the sphingomyelin molecular masses using ESI+ and then their respective LCB-FA combinations(s) using APCI+( )MS/MS. At least 36 protonated molecules of intact sphingomyelin were detected in the bovine milk sample. The combinations found covered a range of molecular masses from 673 to 815 Da. The 12 most common protonated molecules (constituting approximately 90% of the total ion current in ESI) were composed of at least 25 different LCB-FA combinations. Saturated and unsaturated LCBs and FAs were detected in addition to hydroxy fatty acids. The most common LCBs were 16:1, 17:1, 18:1 and 19:1, whereas the most common FAs were 16:0, 22:0, 23:0 and 24:0. LCB-FA combinations of sphingomyelin from bovine brian, bovine erythrocytes and chicken egg yolk are also presented. PMID- 9875524 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the analysis of RNase H cleavage products. AB - RNase H is an endonuclease which cleaves RNA at points of hybridization with DNA. However, certain ambiguities exist in terms of its specificity and location of cleavage along the RNA strand. The analysis of RNase H reaction products of an oligoribonucleotide hairpin by matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI/TOF-MS) is demonstrated. The oligoribonucleotide studied has a highly stable secondary structure which reduces the efficiency of hybridization with the chimeric oligonucleotide used to direct RNase H cleavage. By monitoring the reaction products under different conditions using MALDI/TOF-MS, the optimum variables for cleavage of this highly stable hairpin structure can be determined. PMID- 9875525 TI - Characterization of olanzapine (LY170053) in human liver slices by liquid chromatography/tandem mass spectrometry. AB - Olanzapine metabolism was investigated using incubation of olanzapine with human liver slices. The intent of the investigation was to identify olanzapine metabolites and determine if the human liver slice incubations could potentially produce quantities of the olanzapine glucuronides for future studies. Along with known Phase 1 olanzapine metabolites, N-desmethyl-, 2-hydroxymethyl-, and 4'-N oxide-, a new hydroxylated species was detected. Detection of Phase 2 metabolites included known N-10-glucuronides, a quaternary glucuronide and a novel glucuronide conjugate. This investigation showed the feasibility of using human liver slices to produce sufficient quantities of olanzapine glucuronides for further studies. PMID- 9875526 TI - Investigation of the covalent modification of the catalytic triad of human cytomegalovirus protease by pseudo-reversible beta-lactam inhibitors and a peptide chloromethylketone. AB - An investigation into the interaction between human cytomegalovirus (HCMV) protease and several beta-lactams, with characterization of the resulting acylenzymes using mass spectrometry, is reported. The time dependence of the inhibitors is highlighted by making comparisons of values obtained for inhibition and acylation. Analysis of inactivated HCMV protease revealed a beta-lactam: protease stoichiometry of 1. Subsequent enzymatic digestion with trypsin, peptide mapping using liquid chromatography coupled with electrospray ionization mass spectrometry and sequencing by nanoelectrospray tandem mass spectrometry (NanoES MS/MS) allowed the identification of the site of covalent modification and confirmed Ser 132 as the active site hydroxyl nucleophile. Further, treatment of the protease with a peptide chloromethylketone and sequence analysis using NanoES MS/MS of the alkylated enzyme confirmed His 63 as the active site imidazole nucleophile. PMID- 9875527 TI - Daytime melatonin infusions induce sleep in pigeons without altering subsequent amounts of nocturnal sleep. AB - Daily infusions of melatonin restore sleep suppressed by continuous bright light in pigeons. To test whether melatonin could also induce sleep in pigeons on a 12:12 h light-dark cycle (LD), pigeons received 12-h intravenous melatonin infusions during the day. Melatonin induced sleep during the day, increased EEG slow wave activity, and decreased body temperature and locomotor activity. None of these variables were altered during the night following infusions. The induction of extended daytime sleep by melatonin infusions indicates that melatonin is a principal factor in the regulation of sleep in pigeons. PMID- 9875528 TI - The -491 A/T polymorphism in the regulatory region of the apolipoprotein E gene and early-onset Alzheimer's disease. AB - The -491 polymorphism in the promoter region of the apolipoprotein E gene (APOE) has been suggested to be associated with increased risk for Alzheimer's disease (AD) independent of APOE status. We studied the association between the -491 polymorphism and risk for early-onset Alzheimer's disease in 99 Dutch and 78 Spanish patients. In patients with early-onset AD, we found no consistent relationship with a single allele of the -491 polymorphism. Linkage disequilibrium between the polymorphism and the APOE gene was found which most likely might explain the inconsistent findings. PMID- 9875529 TI - Uric acid inhibits L-DOPA-CU(II) mediated DNA cleavage. AB - It has been proposed that considerable DNA damage may be caused by endogenous metabolites produced during the body's normal metabolic processes. We have previously shown that L-DOPA, in the presence of Cu(II) leads to oxidative DNA breakage in vitro. Uric acid is considered to be a naturally occuring antioxidant and is present in plasma at a relatively high concentration. In this paper we report that uric acid inhibits L-DOPA-Cu(II) mediated DNA cleavage at concentrations similar to or lower than those found in plasma. Xanthine, which is the structural analogue of uric acid is a more potent inhibitor of the reaction. Uric acid was also shown to directly quench the generation of hydroxyl radicals by L-DOPA-Cu(II). The results have been discussed in relation to the putative protective role of uric acid against endogenous DNA damage by oxygen radicals. PMID- 9875530 TI - Remodeling dendritic spines in the rat pilocarpine model of temporal lobe epilepsy. AB - Dendritic degeneration is a common pathology in temporal lobe epilepsy and its animal models. However, little is known when and how the degeneration occurs. In the present study of the rat pilocarpine model, visualization of dendrites of the hippocampal dentate granule cells (DGCs) by biocytin revealed a generalized spine loss immediately after the acute seizure induced by pilocarpine. However, this generalized damage was followed by recovery and plastic changes in spine shape and density, which occurred 15-35 days after the initial acute seizure, i.e., during the period of establishing a chronic phase of this model with the induction of spontaneous seizures. The present finding suggests that initial acute seizures do not cause permanent damages in dendrites and spines of DGCs; instead, dendritic spines are dynamically maintained in the course of the establishment and maintenance of spontaneous seizures. Local dendritic spine degeneration, detected later in the chronic phase of epilepsy, is likely to have a separate cause from initial acute insults. PMID- 9875531 TI - Peripheral axons of Wlds mice, which regenerate after a delay of several weeks, do so readily when transcription is inhibited in the distal stump. AB - We have raised the hypothesis that differentiated Schwann cells repress regrowth of axons but become permissive upon dedifferentiation. WId(S) mouse is a strain in which severed peripheral nerves do not degenerate for several weeks, and axonal regeneration does not occur either [5,11]. In this strain, we studied the role of resident cells upon axonal regeneration by inhibiting transcription. Regeneration was assessed with the pinch test, electron microscopy and Dil (a fluorescent lipid soluble dye). After a crush, WId(S) axons did not regenerate but they did so when the crush was made through a nerve segment treated with actinomycin D (ActD), an inhibitor of transcription. In contrast, when the crush was made distal to the treated segment no regeneration ensued. Our results support the notion that normal resident cells of peripheral nerves repress axonal growth. PMID- 9875532 TI - Degenerative terminals of the perforant pathway are human alpha-synuclein immunoreactive in the hippocampus of patients with diffuse Lewy body disease. AB - We investigated the hippocampal pathology in diffuse Lewy body disease (DLBD) using alpha-synuclein immunohistochemistry. Ubiquitin-positive intrahippocampal structures caused by the degeneration of terminal axons of the perforant pathway were observed to be alpha-synuclein immunoreactive. These alpha-synuclein positive degenerative terminals contained granulo-filamentous or vesiculo-tubular components similar to those of Lewy bodies (LB) immunoelectron microscopically, suggesting that alpha-synuclein may abnormally aggregate into filamentous or membranous cytoskeletal components including neurofilaments and synaptic vesicles in DLBD. A 'dying back' degenerating process due to a blockage of axonal transport may explain why the degenerative terminals and LB share similar alpha synuclein-positive components, but the origin cells of the perforant pathway contain only a few LB. PMID- 9875533 TI - Antiserum against Cl- pump complex recognizes 51 kDa protein, a possible catalytic unit in the rat brain. AB - We purified Cl- pump in the rat brain and obtained 520 or 580 kDa protein complexes which consisted of 62, 60, 55 and 51 kDa proteins. An antiserum against 520 kDa protein complex recognized 51 kDa protein in both 520 and 580 kDa complexes, and reduced both Cl(-)-ATPase and Cl(-) pump activities. Such an immunoreactive 51 kDa protein was found in the brain, spinal cord and kidney. When incubated with [gamma-(32)P]ATP, the protein complex yielded phosphorylated 51 kDa protein, the label being hydroxylamine-sensitive and increased in the presence of Cl- and/or an inhibitor of Cl- pump, ethacrynic acid. Thus, the antibody appears to recognize a possible catalytic subunit of Cl- pump, 51 kDa protein, in the rat. PMID- 9875534 TI - Suppression of inspiratory fast rhythm, but not bilateral short-term synchronization, by morphine in anesthetized rabbit. AB - To better understand the involvement of opioid receptor systems in the respiratory control, effects of morphine on the high-frequency component of inspiratory nerve discharge were evaluated. The inspiratory fast rhythm in the bilateral phrenic nerve discharge of anesthetized and artificially ventilated rabbits was analyzed with power spectral and coherence functions. Morphine (0.625 10 mg/kg, i.v.) decreased the amplitude and frequency of the inspiratory fast rhythm in a dose-dependent manner via naloxone-sensitive mechanisms. In contrast, the bilateral short-time scale correlation of the phrenic fast rhythm was resistant to morphine even at a strong suppression of the respiratory activities. It is concluded that there is little influence of opioid receptor system to the neural connectivity underlying bilateral phrenic synchronization. PMID- 9875535 TI - Circadian rhythm and light responsiveness of BMAL1 expression, a partner of mammalian clock gene Clock, in the suprachiasmatic nucleus of rats. AB - To clarify whether BMAL1 is involved in the photic signal transduction in the mammalian circadian clock, we examined the effects of a single light pulse on the level of BMAL1 mRNA in the suprachiasmatic nucleus (SCN) of rats by in situ hybridization. Rats were exposed to 30 min light of ca. 300 lux at six different phases in constant darkness and decapitated 60 min later. BMAL1 transcripts in the SCN of the control animals showed a robust circadian oscillation with the highest expression at ZT (Zeitgeber time) 18 and the lowest at ZT2. The light pulse slightly increased the level of BMAL1 transcripts in the SCN. However, the increment did not depend on the phase of light pulse. There was no significant change in the BMAL1 mRNA level up to 120 min after a light pulse at ZT14 and ZT22. These results indicate that BMAL1 transcription is not involved in the photic signal transduction responsible for non-parametric entrainment of the circadian clock in rats. PMID- 9875536 TI - Postnatal changes in Bax-immunoreactivity and apoptosis of the rat trigeminal primary neurons. AB - The developmental changes of Bax protein-like immunoreactivity (Bax-ir) and naturally occurring cell death were investigated in the trigeminal primary neurons of rats, in the first 2 postnatal weeks. At 1 day postpartum, the trigeminal primary neurons exhibited intense cytoplasmic Bax-ir. A densitometric analysis indicated >95% of cells exhibited an ir-density (the staining intensity as compared to the background level) higher than 1.5. By 2 weeks the ir-density significantly decreased with >95% lower than 1.5, that was similar to the adult level. A nick-end labeling method revealed DNA fragmentation in apoptotic trigeminal primary neurons in newborn rats. During the first 11 days the percentage of apoptotic cells was highly correlated to the postnatal days with a regression line y = -0.083x + 0.877 (r = 0.976). A possible role of Bax in neonatal rat primary neurons is discussed. PMID- 9875537 TI - Transcription factor binding to the core promoter of the human monoamine oxidase B gene in the cerebral cortex and in blood cells. AB - Many studies show that monoamine oxidase B in blood cells is a biological marker for personality characteristics such as sensation seeking. The mechanism underlying this association is so far not explored. In the present study we have performed electrophoretic mobility-shift assays to investigate the pattern of protein binding to a 150 bp fragment of the proximal 5'-flanking region of the human monoamine oxidase B gene. We compared the pattern using nuclear extracts from human brain and lymphocytes. Interestingly, a correlation was observed between monoamine oxidase B enzyme activity in blood cells (platelets) and the binding pattern of two uncharacterized transcription factors. These data are well in line with the long-standing notion that interindividual differences in platelet monoamine oxidase may represent differences in expression of the enzyme rather than genotypic variation. PMID- 9875538 TI - Hemispheric differences in frequency dependent dipole orientation of the human auditory evoked field component N100m. AB - Auditory evoked fields (AEF) of 19 healthy male subjects were recorded bilaterally with a Philips 31 -channel biomagnetometer, using two conditions of stimulation (1000 vs. 5000 Hz tones). The N100m latency was characterized by a single moving dipole for each condition and hemisphere using a boundary element model (BEM) as volume conductor. While the right hemispheric dipole orientations and locations did not change with respect to condition, the left hemispheric dipoles differed significantly between the 1000 and 5000 Hz tones, especially in dipole orientation. The left hemispheric dipoles were orientated on average 10.8 degrees more vertically for the 5000 Hz condition. This result points to interhemispheric differences on the level of sensory processing. PMID- 9875539 TI - Evidence for a neurotransmitter role of glutamate in guinea pig myenteric plexus neurons. AB - Longitudinal muscle-myenteric plexus strips of the guinea pig ileum were used to investigate a possible transmitter role of glutamate (Glu) in the enteric nervous system. Glu was released from this nerve muscle preparation by high K(+) depolarization in a Ca2(+)-dependent manner, by electrical field stimulation and by the ganglionic stimulant dimethylphenylpiperazinium which indicates its neural origin. Contractions of the longitudinal muscle evoked by electrical stimulation of the myenteric nerves or by Glu, were significantly reduced by the N-methyl-D aspartate (NMDA)-receptor antagonist FR115427 (9 and 18 microM), whereas contractions induced by histamine were unaffected. The results show that the amino acid Glu is likely to play an excitatory neurotransmitter role via NMDA receptors in the myenteric plexus of the guinea pig. PMID- 9875540 TI - Alz-50/Gallyas-positive lysosome-like intraneuronal granules in Alzheimer's disease and control brains. AB - The Gallyas-Braak silver impregnation method revealed neurons containing well defined intraneuronal granules in both Alzheimer's disease and normal control brains. The granules were immunostained prominently with the Alz-50 antibody and, to a lesser degree, with the tau-2 antibody, but not with other anti-tau antibodies examined. The areas of distribution of granule-containing neurons detected by the Gallyas-Braak method appeared to overlap with the reported main sites of subcortical distribution of neurofibrillary tangles. They, however, were not observed in the cerebral cortex, including the hippocampal region. The Alz-50 immunoreactive granules showed ultrastructural features similar to those of lysosomes or lipofuscin. These findings suggest that denatured tau might be degraded in lysosomes. PMID- 9875541 TI - Activation of the ventral and mesial frontal cortex of the monkey by self initiated movement tasks as revealed by positron emission tomography. AB - In order to investigate the neural mechanisms of movement initiation, we measured the regional cerebral blood flow (rCBF) of the monkey during self-initiated and visually-initiated hand movement tasks using positron emission tomography (PET). The orbitofrontal, cingulate, and anteromedial part of the dorsal premotor areas were preferentially activated by the self-initiated hand movement task (SELF). The pre-supplementary motor area and the cingulate motor area were also included in the active foci during the task. In the visually-initiated task (VISUAL), the V1, V2, V3, V3A, and V4 were activated, whereas the activity of the dorsolateral premotor and primary motor areas was not significantly different between the two tasks. These findings suggest that the orbitofrontal and mesial frontal cortices play an important role in the neural processes involved in self-initiation of movement and self-regulation of inner drives. PMID- 9875542 TI - The N-terminal PDZ-containing region of postsynaptic density-95 mediates association with caveolar-like lipid domains. AB - Postsynaptic density-95 (PSD-95) is a palmitoylated peripheral membrane PDZ protein that organizes signaling molecules at synaptic sites. Here we find the amino terminal region of PSD-95, containing the three PDZ domains is necessary and sufficient to localize PSD-95 to caveolar-like domains. In transfected COS cells, a subpopulation of PSD-95 is buoyant in sucrose gradients, and co-migrates with caveolin, a marker for caveolar domains. Sucrose gradient separation of brain extracts showed that some neuronal PSD-95 protein is present in buoyant fractions as well. Analysis of truncated forms of the PSD-95 indicated that the N terminal PDZ-containing region localizes to caveolae, but the C-terminal region, containing the SH3 and the guanylate kinase domains does not. The mechanism by which the N-terminal region targets PSD-95 to buoyant lipid domains remains unknown. PSD-95 does not interact with caveolin and palmitoylation of PSD-95 is not required for caveolar fractionation. PMID- 9875543 TI - Calcium signals monitored from leopard frog optic tectum after the optic nerve has been selectively loaded with calcium sensitive dye. AB - We loaded adult leopard frog optic nerves with the calcium-sensitive dye Calcium Green-1 3000 mw dextran conjugate. The dye was transported to the optic tectum in approximately 6 days and selectively labeled optic nerve terminals as seen with confocal microscopy. Viewed with an intensified CCD system, electrical stimulation of the optic nerve in vitro increases Calcium Green-1 fluorescence significantly. With increasing number of pulses in pulse trains there was increased presynaptic facilitation as measured by increased fluorescence. Addition of nicotine to the bathing solution increased baseline fluorescence. These results suggest that Calcium Green-1 dextran conjugate can be actively transported in adult nerve fibers over a significant distance and is retained in presynaptic terminals in a form that allows monitoring of presynaptic calcium levels. PMID- 9875544 TI - Organization of the forepaw representation in ferret somatosensory cortex. AB - Little is known about the function and structure of ferret somatosensory cortex. We used anatomical methods and multi-unit recordings to characterize the cytoarchitecture, functional responses and topography of the forepaw representation in ferret somatosensory cortex. The representation of the cutaneous ferret forepaw encompasses approximately the caudal half of the posterior sigmoid gyrus. The posterior sigmoid gyrus and coronal sulcus contain unique cytoarchitectonic fields that conform in large part to earlier descriptions of somatosensory regions in the cat. The cytoarchitectonic regions form mediolateral bands that comprise areas 4, 3a, 3b, 1, and 2 (from rostral to caudal). We studied most extensively areas 3a and 3b for functional responses to somatic stimuli; our data indicate that ferret somatosensory cortex contains at least two representations of the forepaw in these two areas. Our data also suggest that within ferret somatosensory cortex, the morphological and submodality features gradually, rather than abruptly, distinguish themselves as unique cortical fields. PMID- 9875545 TI - "Warmth-insensitive fields": evidence of sparse and irregular innervation of human skin by the warmth sense. AB - Although more acute in some areas of the body than in others, temperature sensitivity is assumed to be present throughout the skin. Only when very small stimuli have been used (e.g., approximately 1 mm2) has sensitivity to warming or cooling appeared discontinuous. Here we report the discovery of patches of skin several square centimeters in area within which heating cannot be detected until skin temperature exceeds the thresholds of C heat-sensitive nociceptors (>41 degrees C). These warmth-insensitive fields (> or = 5 cm2), which appear to lack low-threshold warm fibers, were also found to have reduced responsiveness to non painful heating and significantly higher heat pain thresholds compared to surrounding areas of skin. The existence of such sites corroborates reports that warm fibers are rare in human cutaneous nerves and confirms the classical theory that cutaneous innervation by the warmth sense is punctate and sparse. The insensitive areas also provide unique opportunities for assessing the contribution of the low-threshold warmth system to perception of heat and heat pain, and their existence in healthy young adults contraindicates use of warmth sensitivity in neurological assessments of C-fiber function. PMID- 9875546 TI - Distribution of inhibitory synapses on the somata of pyramidal neurons in cat motor cortex. AB - The mechanisms by which cortical neurons perform spatial and temporal integration of synaptic inputs are dependent, in large part, on the numbers, types, and distributions of their synapses. To further our understanding of these integrative mechanisms, we examined the distribution of synapses on identified classes of cortical neurons. Pyramidal cells in the cat motor cortex projecting either to the ipsilateral somatosensory cortex or to the spinal cord were labeled by the retrograde transport of horseradish peroxidase. Entire soma of selected corticocortical and corticospinal cells were examined using serial-section electron microscopy. The profiles of these somata and the synapses formed with each of these profiles were reconstructed from each thin section with a computer aided morphometry system. All somatic synapses were of the symmetrical, presumably inhibitory type. For both cell types, these synapses were not homogeneously distributed over the somatic membrane, but were clustered at several discrete zones. The number and density of synapses on the somata of different corticocortical and corticospinal neurons were not significantly different. However, the density of these synapses was inversely correlated with the size of their postsynaptic somata. We discuss the significance of these findings to the integrative properties of cortical neurons. PMID- 9875547 TI - Neuronal activity in the primate supplementary motor area and the primary motor cortex in relation to spatio-temporal bimanual coordination. AB - Single neuronal activity was recorded from the supplementary motor area (SMA proper and pre-SMA) and primary motor cortex (M1) in two Macaca fascicularis trained to perform a delayed conditional sequence of coordinated bimanual pull and grasp movements. The behavioural paradigm was designed to distinguish neuronal activity associated with bimanual coordination from that related to a comparable motor sequence but executed unimanually (left or right arm only). The bimanual and unimanual trials were instructed in a random order by a visual cue. Following the cue, there was a waiting period until presentation of a "go signal", signalling the monkey to perform the instructed movement. A total of 143 task-related neurons were recorded from the SMA (SMA-proper, 62; pre-SMA, 81). Most SMA units (87%) were active in both unimanual contralateral and unimanual ipsilateral trials (bilateral neurons), whereas 9% of units were active only in unimanual contralateral trials and 3% were active only in unimanual ipsilateral trials. Forty-eight per cent of SMA task-related units were classified as bimanual, defined as neurons in which the activity observed in bimanual trials could not be predicted from that associated with unimanual trials when comparing the same events related to the same arm. For direct comparison, 527 neurons were recorded from M1 in the same monkeys performing the same tasks. The comparison showed that M1 contains significantly less bilateral neurons (75%) than the SMA, whereas the reverse was observed for contralateral neurons (22% in M1). The proportion of M1 bimanual cells (53%) was not statistically different from that observed in the SMA. The results suggest that both the SMA and M1 may contribute to the control of sequential bimanual coordinated movements. Interlimb coordination may then take place in a distributed network including at least the SMA and M1, but the contribution of other cortical and subcortical areas such as cingulate motor cortex and basal ganglia remains to be investigated. PMID- 9875548 TI - Cutaneous texture discrimination following transection of the dorsal spinal column in monkeys. AB - Transection of the dorsal spinal column in monkeys has been shown to impair discrimination of the frequency or duration of repetitive tactile stimulation, without recovery over extended periods of postoperative testing. These deficits would be likely to prevent discrimination between textures presented passively and in sequence, if repetitive temporal sequences were distinguishing features of the textures. However, previous investigations of texture discrimination after dorsal column section did not obtain a deficit on tests involving active palpation of sandpaper surfaces. In the present study, rows of raised dots were stroked across the glabrous skin of one foot so that temporal entrainment of neural activity would constitute a prominent cue. The rows were oriented mediolaterally, and the textures moved proximodistally across the skin surface (varying the spacing between the rows). Four monkeys were trained to release a lever when the rougher of two textures was in contact with the skin, and the rough texture was preceded by one to three passes of a smooth texture. Stable levels of preoperative performance ranged from 78.6 to 85.7% correct responses. After interruption of the ipsilateral dorsal column, each monkey was impaired over at least 2 months of testing. One animal did not show evidence of recovery; two recovered partially from the initial deficit; and one returned to preoperative levels of performance after extensive retraining. These results are interpreted in terms of aberrant inhibitory influences which result from repetitive stimulation after a dorsal column lesion. PMID- 9875549 TI - Vibrotactile stimulus order effects in somatosensory cortical areas of rhesus monkeys. AB - This study demonstrated effects of stimulus order on single-cell responses in the macaque primary and secondary somatosensory and 7b cortical areas. As part of a study of tactile attention, two monkeys (Macaca mulatta) received similar constant amplitude, sinusoidal tactile vibration patterns (125 Hz) at two glabrous skin, hand locations. The stimuli started asynchronously with offsets of 150 or 300 ms. In cells with bilateral receptive fields and increased firing to a stimulus, we observed an average lowering of 30% in the firing rates to the contralateral stimulus when preceded by stimulation of the ipsilateral hand. Some cells with only contralateral receptive fields showed similar depressed responses to a contralateral stimulus when preceded by an ipsilateral stimulus. Stimulus order effects were more prominent during dual stimulation of the receptive field on one hand. In six cells whose background activity was inhibited by the first stimulus, higher rates appeared at the onset of the second stimulus. These results suggest a possible substrate for psychophysical findings of stimulus masking in which a preceding stimulus depresses detection thresholds. The spatial and temporal characteristics of in-field inhibitory mechanisms best account for the observed stimulus order effects. PMID- 9875550 TI - Strychnine- and bicuculline-induced changes in the firing pattern of motoneurones during in vitro fictive locomotion reveal a possible N-methyl-D-aspartic acid (NMDA)-mediated suppression of motor discharge in wallaby (Macropus eugenii) pouch young. AB - Inhibitory pathways in the spinal cord play an important role in establishing the pattern of motor discharge. In the wallaby spinal cord preparation, disruption of glycinergic and gamma-amino butyric acid (GABA)ergic neurotransmission abolished the alternation between antagonistic motor pools during fictive locomotion. A new pattern of motor discharge also appeared when both glycine and GABA(A) receptors were blocked simultaneously. This discharge pattern was biphasic, characterized by a distinct pause between two bursts of motoneurone firing during each cycle of motor activity. Whole cell patch recordings showed that the second burst of motor discharge was not caused by a separate inward current at a delayed time course. Furthermore, local injection of an N-methyl-D-aspartic acid (NMDA) specific antagonist converted the biphasic discharge to a continuous burst pattern. The result suggests an NMDA-mediated mechanism, which causes a suppression of motoneurone firing when glutamate release from interneurones is enhanced in the absence of glycinergic and GABAergic inhibition. PMID- 9875551 TI - Role of the liver in interorgan homeostasis of glutathione and cyst(e)ine. AB - The most widely recognized function of reduced glutathione (GSH) is its defense against toxic compounds, whether exogenous, such as electophilic xenobiotics, or endogenous, such as reactive oxygen species, generated during normal oxidative metabolism and/or stress. However another no less significant role of GSH-namely its function as a reservoir and vehicle for packaging and transport of cyst(e)ine has been receiving increasing attention. Because GSH is relatively more auto oxidation resistant and stable than cyst(e)ine (CYSH), it serves as the preferred form for storage and transport of the latter especially in the extracellular and relatively much less reduced (than intracellular) milieu, where CYSH oxidizes to cystine (CYSS) rapidly. Over the past two decades, significant work has been going on to delineate the intra- and extrahepatic (interorgan) turnover, transport, and disposal of GSH and define the quantitative role of these processes in interorgan homeostasis of GSH, CYSH, and CYSS. These studies have identified the liver as the central organ of interorgan GSH homeostasis, with sinusoidal GSH efflux as the major determinant of plasma GSH, CYSH, CYSS, and thiol-disulfide status of plasma. This article focuses on the principal components and determinants of interorgan homeostasis of GSH and its breakdown products. It also presents the current state of knowledge under both normal and diseased states. PMID- 9875552 TI - Regulation of hepatic glutathione synthesis. AB - Glutathione (GSH) is one of the most important intracellular peptides, playing a multifunctional role ranging from antioxidant defense to modulation of immune function. GSH is synthesized by all mammalian cells, and the synthesis of GSH is a tightly regulated process. Two of the major determinants of GSH synthesis are the availability of cysteine, the sulfur amino acid precursor, and the activity of the rate-limiting enzyme, gamma-glutamylcysteine synthetase (GCS). In the liver, the major factors that determine the availability of cysteine are the activities of the membrane transport processes of the three sulfur amino acids- cysteine, cystine (under certain oxidative stress conditions) and methionine--and the conversion of methionine to cysteine through the trans-sulfuration pathway. Since the molecular cloning of GCS, there has been an explosion of knowledge regarding how this enzyme is regulated. Both transcriptional and posttranscriptional regulation play important roles in modulating the activity of this critical cellular enzyme. PMID- 9875553 TI - Human glutathione S-transferases. AB - Human glutathione S-transferases (GSTs) are a functionally diverse family of soluble enzymes of detoxification that use reduced glutathione (GSH) in conjugation and reduction reactions. Toxic electrophiles, including a variety of carcinogens, are substrates for the GSTs and after conjugation or reduction they are more easily excreted into bile or urine. Many of the GSTs have been cloned, and the three-dimensional structures of GSTs from several species, including humans, have been determined. These data have provided significant insight into how the GSTs function as enzymes. Many GST substrates are inducers of GST gene expression; nonsubstrate inducers include H2O2 and other reactive oxygen species. The regulatory elements of several human GST genes have been partially characterized, and the regulation of the GSTs in humans appears to be very different from that in rodents. Several polymorphisms of GST expression occur commonly in humans and have been associated with an increased susceptibility to certain cancers, particularly when combined with other genetic and environmental factors such as smoking. The role of GSTs in protecting cells from injury by toxic electrophiles continues to be developed. PMID- 9875554 TI - Excretion of GSSG and glutathione conjugates mediated by MRP1 and cMOAT/MRP2. AB - It has been shown that both multidrug resistance-associated protein (MRP1) and canalicular multispecific organic anion transporter (cMOAT/MRP2) have the ability to extrude glutathione conjugates (GS-X pump activity)from cells. Therefore, they play an important role in the detoxification of xenobiotics. Using mrp1-knockout mice, it has recently been shown that MRP1/mrp1 has an important role in the export of leukotriene C4 (LTC4), a mediator of inflammation, and in protecting the body from a number of toxins, including several antitumor drugs. A comparison of the transport properties across the bile canalicular membrane in normal and mutant rats, whose cMOAT function is hereditarily defective, has shown that the physiologic role of cMOAT is to excrete LTC4, bilirubin glucuronides, 171 estradiol-170-D-glucuronide, and reduced folates. In the present review article, we summarize the substrate specificity and mechanism for the transport of these GS-X pumps, focusing on the pharmacologic and physiologic aspects. The transport activity mediated by cMOAT is also discussed in terms of a comparison between membrane vesicles from hepatocytes and cMOAT-transfected cells, and we also briefly examine the possible role of MRPI and cMOAT in the extrusion of reduced glutathione. PMID- 9875555 TI - Roles of MRP2 and oatp1 in hepatocellular export of reduced glutathione. AB - Transport of reduced glutathione (GSH) into the extracellular space is the initial and perhaps limiting step in the turnover of the tripeptide in all mammalian cells; however, the transport system or systems that mediate GSH efflux remain obscure. In the liver, a major site of GSH synthesis, GSH is released at high rates into both blood plasma and bile. Nearly half of the GSH released by rat hepatocytes is transported across the canalicular membrane into bile, with biliary GSH concentrations reaching 8 to 10 mM. GSH transport into bile functions as a driving force for bile secretion and plays an important role in hepatic detoxification of drugs, metals, and other reactive compounds of both endogenous and exogenous origin. The remainder of the GSH is released across the sinusoidal membrane into blood plasma for delivery to other tissues. The molecular mechanisms of GSH efflux have not been identified for any cell type, although recent studies provide important insight into possible mechanisms. In particular, oatp1, the sinusoidal organic solute transporter, was recently shown to function as a GSH/organic solute exchanger. This finding identifies both the energy coupling mechanism for oatp1 and a pathway for GSH release into blood plasma. However, oatp probably only accounts for a fraction of the total GSH released into sinusoidal blood. A candidate canalicular GSH transport mechanism has also recently been described. Canalicular GSH efflux may be mediated by the adenosine 5'-triphosphate (ATP)-dependent organic solute transport protein MRP2 (also termed cMOAT or cMRP). MRP2 is a member of the multidrug resistance-associated family of proteins (MRP) whose preferred substrates include glutathione S conjugates. Recent studies suggest that MRP can also transport GSH itself. This report summarizes the evidence documenting a role for oatp1 and MRP2 in GSH efflux from hepatocytes, and their possible contribution to hepatic GSH homeostasis. PMID- 9875556 TI - Mitochondrial glutathione: importance and transport. AB - Accumulating evidence pointing to mitochondria as critical participants in the control of apoptotic and necrotic cell death and in the development of specific disease states has led to a renaissance on the study of these organelles. Because mitochondria are the major consumers of molecular oxygen within cells, they stand as one of the most important generators of reactive oxygen species and therefore constitute potential targets of therapeutic intervention in pathologic states in which oxidative stress originates from these organelles. In this regard, mitochondria are specific targets of ethanol intoxication, thereby leading to reported morphologic and functional alterations of mitochondria. Because mitochondria are also indispensable for the maintenance of cell functions, their dysfunction induced by ethanol may be a key event in the development of alcoholic liver disease. Indeed, chronic ethanol feeding in experimental animals has been reported to cause a selective deficiency in the availability of reduced glutathione (GSH) in mitochondria due to the impaired functioning of the specific mitochondrial carrier that translocates GSH from cytosol into the mitochondrial matrix. Such a selective depletion sensitizes hepatocytes from chronic ethanol fed animals to the oxidative effects of cytokines, e.g., tumor necrosis factor (TNF). Restoration of mitochondrial GSH by the in vivo administration of S adenosyl-L-methionine or the in vitro use of GSH ethyl ester prevents the susceptibility of hepatocytes to TNF. Although the nature of this specific carrier has not yet been uncovered, the elucidation of the mechanisms whereby ethanol leads to its impaired activity may provide important clues as to its function and mechanism of action, which in turn may be useful toward the definitive characterization and identification of this important carrier. PMID- 9875557 TI - Glutathione defense in non-parenchymal cells. AB - Toxicity to nonparenchymal cells can result in disruption of the hepatic microcirculation, altered production of cytokines, and hepatic fibrosis. Many of the relevant insults produce oxidative stress or toxic metabolites that require glutathione detoxification. This article reviews the role of sinusoidal endothelial cell glutathione (GSH) in reperfusion injury, cytomegalovirus infection, and hepatic venoocclusive disease. The effects of oxidative stress and antioxidants on Kupffer cell production of cytokines and, in particular the potential benefit of antioxidants in the setting of reperfusion injury, are discussed. Oxidative stress upregulates collagen gene expression by stellate cells, and this is modulated by antioxidants. Current thinking on intrahepatic GSH and cysteine homeostasis is discussed. Finally, I review the published data on nonparenchymal GSH levels, glutathione S-transferase activity and isoenzyme pattern, and glutathione peroxidase activity. PMID- 9875558 TI - Glutathione therapy: from prodrugs to genes. AB - Glutathione (GSH; L-gamma-glutamyl-L-cysteineglycine) is found in almost all mammalian cells, and liver has very high intracellular levels of GSH. It has many cellular functions, such as being a coenzyme, maintaining thiol/disulfide status, protection against toxic compounds and oxidative stress. GSH levels have been reported to be low in a number of pathological conditions; thus methods for increasing GSH levels are desirable. GSH may be increased by supplying its amino acid precursor cysteine, in the form of prodrugs, such as N-acetylcysteine (NAC) and 2-oxothiazolidine-4-carboxylate (OTC). It may also be increased by giving gamma-glutamylcysteine, a dipeptide precursor GSH monoester and GSH diester are effective GSH delivery drugs. Such compounds may be therapeutically useful. Gene therapy may be useful for longer term therapy of GSH deficiency. PMID- 9875559 TI - The treatment of erythropoietic protoporphyria. PMID- 9875560 TI - Kainic acid may enhance hippocampal NO generation of awake rats in a seizure stage-related fashion. AB - Temporal changes in kainate-induced seizure activity and hippocampal NO generation were evaluated simultaneously in conscious rats by using in vivo microdialysis and by determining the concentrations of nitrite and nitrate in perfusates. Intraperitoneal injection of kainic acid produced 'wet dog shake', focal seizure of the limbs and neck, hypersalivation, or generalized convulsion. These behavioral changes peaked at 120 min after the drug challenge and lasted for about 100 min. In contrast, the concentrations of NO metabolites, nitrite and nitrate, in the hippocampal perfusates increased rapidly and reached a plateau level at 40 min after the injection, and the level remained high for over 220 min. The increase was more marked in animals presenting severe seizures than those presenting mild ones. Pre-treatment with 25 mg/kg N(G)-nitro-L-arginine methyl ester promoted the severity of kainate-induced seizures, but it suppressed the increase in NO metabolites. These results suggest that kainic acid enhances hippocampal NO generation in a severity-related manner of the induced seizures. The enhanced NO generation upon kainate challenge appears mainly to be involved in seizure suppression. PMID- 9875561 TI - Chronic cyclosporine-A injection in rats with damaged blood-brain barrier does not impair retention of passive avoidance. AB - Recently, we demonstrated that chronic administration of immunosuppressant drug, cyclosporine-A (CsA), does not produce impairment in memory retention of a passive avoidance task in normal adult rats. Since CsA has been used as an adjunctive therapy to avoid xenograft rejection inherent in neural transplantation therapy for neurodegenerative disorders, we replicated our previous study in animals with damaged blood-brain barrier (BBB) simulating that of the neural transplantation protocol. Adult rats with damaged BBB that received either chronic CsA (5, 10, and 20 mg/kg) or vehicle injection did not differ significantly in their memory retention of the passive avoidance task that rewarded 'less mobile activity', in that animals avoided electric shock when they restrained their movements within the safe compartment. General spontaneous locomotor activity also was not altered by CsA, except in animals that received 20 mg/kg, which displayed significant hypoactivity at later post-injection periods of CsA. The absence of potentiation of retention of the passive avoidance task in all CsA-treated animals, including the hypoactive ones, suggests that locomotor activity did not interfere with cognitive behavior. The present results confirm our previous findings that the therapeutic dosage (10 mg/kg) of CsA used for neural transplantation does not produce visible deleterious effects on the performance of memory retention task in immunosuppressed rats with damaged BBB. PMID- 9875562 TI - Relationships between glutamate release, blood flow and spreading depression: real-time monitoring using an electroenzymatic dialysis electrode. AB - Spreading depression (SD) in a flow-restricted area of the brain may be prolonged and may become potentially harmful by releasing glutamate. We induced SD in an oligemia model and examined the subsequent glutamate release. In 18 anesthetized male Fischer rats, a laser Doppler flowmeter, an electroenzymatic electrode for continuous measurement of glutamate, and a calomel electrode for measuring DC potential were placed through a cranial window positioned 3 mm away from a second window where KCl-soaked cotton was placed to initiate SD. The left carotid artery or both the common carotid arteries were ligated to suppress reactive hyperemia of SD. SD produced an increase in glutamate from 24.8 +/- 13.8 to 33.5 +/- 25.3 microM (peak value) (P < 0.0001). After ligation of both carotid arteries, the duration of SD increased from 1.5 +/- 0.6 min (before ligation) to 6.4 +/- 5.1 min (P < 0.05). Glutamate reached a peak level of 63.9 +/- 72.3 microM, then quickly returned to the control value. However, there was no positive correlation between the duration of SD and glutamate concentration. It is concluded that prolonged SD is not accompanied by a progressive increase in glutamate. Therefore, glutamate release induced by SD may not exert harmful effects on neurons. PMID- 9875563 TI - Very slow oscillatory activities in lateral geniculate neurons of freely moving and anesthetized rats. AB - In urethane anesthetized rats many lateral geniculate neurons display a strong very slow oscillatory behavior in the range of 0.025-0.01 Hz. One of the aims of the present study was to determine whether very slow oscillatory activity in this range can also be obtained in barbiturate anesthetized and in awake animals, respectively. Although very slow oscillations were found in geniculate neurons both during awakeness and during anesthesia, significant differences in peak frequencies of oscillations under the three experimental conditions (barbiturate, urethane, awake) were demonstrated. In addition, we have tested the influence of glutamate antagonists and GABA agonists as well as antagonists on the very slow oscillatory activity in urethane anesthetized rats. Very slow oscillatory activity which could be blocked by the continuous illumination of the eyes was re induced by iontophoresis of NMDA and non-NMDA glutamate antagonists. GABA(A) as well as GABA(B) agonists also caused a significant re-induction of very slow oscillatory activity under light conditions. In the dark, muscimol, a GABA(A) agonist, significantly enhanced the very slow oscillatory activity, i.e. muscimol either induced it or reduced the frequency of very slow oscillations. For the whole sample, GABA antagonists did not have a significant influence on the very slow oscillatory activity. Autocorrelation analysis based on the spike interval histograms and determination of the spectrum of autocorrelograms revealed the significance of periodicity. PMID- 9875564 TI - The expression and localization of serine proteinase inhibitor PI-6 mRNA in developmental and ischemic mouse brain. AB - A serine proteinase inhibitor, PI-6, is a member of the serine proteinase inhibitor (serpin) superfamily. In the present study, we investigated the developmental expression of PI-6 in the mouse brain and the effect of experimental ischemia on the expression of PI-6 in the adult brain. Northern blot analysis showed a high level of expression of PI-6 mRNA in brain stem and diencephalon as compared with other regions in the adult brain. The expression of PI-6 mRNA in the whole brain was increased gradually until 11 days after birth and was decreased again in the adult brain. In situ hybridization analysis revealed that the mRNA was localized in pyramidal cell layer of the post-natal hippocampus, especially in CA3 region, and in layer V of the cerebral cortex. In the brain stem, two specific motor nuclei, the facial nucleus and the motor nucleus of trigeminal nerve, which are important to active feeding, were strongly positive for PI-6 mRNA. Brain ischemia induced by bilateral ligation of the common carotid artery led to an increase in PI-6 mRNA expression in the whole brain, accompanied by the degeneration of hippocampal pyramidal cells. These results indicate distinct temporal and spatial expression of PI-6 in the mouse brain and suggest the involvement of PI-6 in the maturation of neurons and degenerative and regenerative processes. PMID- 9875565 TI - Nerve growth factor increases sensitivity to bradykinin, mediated through B2 receptors, in capsaicin-sensitive small neurons cultured from rat dorsal root ganglia. AB - To examine the effects of nerve growth factor (NGF) on the response to bradykinin (BK) of primary afferent neurons, intracellular recordings were obtained from small (< 30 microm) and large (> or = 35 microm) neurons in rat dorsal root ganglia (DRG). The response to BK in the small neurons was tested in 23 freshly dissociated neurons (dissociated group), 37 neurons cultured in the absence of NGF (no-NGF group) and 117 neurons in the presence of NGF (NGF group). Application of BK (10(-7) or 10(-5) M) induced a depolarization in a small number of neurons in the freshly dissociated (13%) and the no-NGF (11%) groups. After cultivation with NGF, the percentage of neurons that were depolarized by BK significantly increased to 46% after 2 days of cultivation. In the NGF group, the percentage of neurons sensitive to BK was significantly greater among capsaicin (CAP)-sensitive than among CAP-insensitive neurons (48 vs 20%). This BK-induced depolarization was completely blocked by a B2 receptor antagonist, but not a B1 receptor antagonist. With large neurons, in contrast, NGF did not increase the percentage that were BK-sensitive (9% in the dissociated group vs 0% after being cultured 2 days with NGF). These results demonstrate that NGF increases sensitivity to BK, mediated through B2 receptors only, in capsaicin-sensitive small neurons cultured from rat DRGs. PMID- 9875566 TI - Direct projections from the entorhinal cortical layers to the dentate gyrus, hippocampus, and subicular complex in the cat. AB - Projections from each layer of the entorhinal cortex (EC) of the cat were traced to the dentate gyrus (DG), Ammon's horn (CA), prosubiculum (ProSb), subiculum (Sb), presubiculum (PreSb) and parasubiculum (ParaSb); the anterograde or retrograde labeling method was used after stereotaxic injection of wheat germ agglutinin-horseradish peroxidase, cholera toxin B subunit, or Phaseolus vulgaris leucoagglutinin. On the side ipsilateral to the tracer-injection, layer II of the EC projected most abundantly to the outer half of the molecular layer (ML) of the DG, less abundantly to the almost entire thickness of the stratum lacunosum moleculare (SLM) of CA2-3, moderately to the almost entire thickness of the SLM of CA1, and less to the outer part of the ML of the ProSb and Sb. Layer III projected abundantly to the almost entire thickness of the SLM of CA1 and outer part of the ML of the ProSb and Sb, and sparsely to the SLM of CA2-3. Layer IV projected sparsely to the pyramidal cell layer of the ProSb and Sb; Layer IV of the medial part (toward the ParaSb) of the EC projected further to the ML of the DG. Layer VI projected sparsely to the outer part of the ML of the DG, almost entire thickness of the SLM of CA1-3, and outer part of the ML of the ProSb and Sb. More temporal parts of the hippocampal region received the projections from progressively more medial and more rostral parts of layers II and III, and from progressively more rostral parts of layers IV and VI. The ML of the PreSb and ParaSb received projections from all layers of the medial part of the ipsilateral EC. The SLM of CA1 and ML of the ProSb, Sb and ParaSb received projections from layer II and/or III of the contralateral medial entorhinal area. PMID- 9875567 TI - Cerebellar control of the cardiovascular responses during postural changes in conscious rabbits. AB - In conscious control rabbits, tilting the head 30 degrees up from a position 30 degrees down induced initially an inhibition in the renal sympathetic nerve activity (RSNA), however this inhibition immediately released and became a transient increase. Following these responses in RSNA, blood pressure (BP) initially decreased but recovered to the control level within 3-5 s. After bilateral destruction of the lateral nodulus-uvula in the cerebellum, in contrast, the same tilting of the head caused an immediate large increase in RSNA without early inhibition, which was sustained at a high level. BP increased transiently, but then decreased and remained at a level lower than the control. These results indicate that the timing and duration of this transient increase in RSNA during tilting the head up are controlled by the lateral nodulus-uvula and may be important in the rapid adaptation of blood pressure. In addition, this suggests that the lateral nodulus-uvula may play an important role in the cardiovascular control under conditions of consciousness during changes in head position and body posture. PMID- 9875568 TI - Responses in the motor cortex time-locked to the sensory stimuli conditioning target-reaching in the cat. AB - Single unit activity in the forelimb area of the sensorimotor cortex (area 4gamma) was investigated in unrestrained cats during target-reaching with the contralateral limb to a morsel of food. An acoustic stimulus (tone) was used as a cue for movement initiation. A total of 159 cells which modulated their activity in relation to the task were investigated and the responses were analysed with respect to the onset of the stimulus and the movement, respectively. Eighteen per cent of the recorded cells showed responses to the conditional signal with a latency ranging from 20 to 100 ms. The response pattern of these neurones always had a second component coupled to the movement. Correlation between timing of neuronal responses and movement onset revealed that the short latency responses were strongly time-locked to the sensory cue but not to the movement. A Go/No-go discrimination task in which the animal was required to initiate or withhold the movement depending on the frequency of the tone showed that the short latency responses were selective to the cue relevant for initiation of the movement and were substantially decreased or absent when a No-go cue was presented. It was also shown that these neurones did not respond to an indifferent cue, but acquired short-latency responses when it was repeatedly rewarded. The results indicate that some motor cortical neurones are recruited in the early stages of sensory-to-motor transformation related to the processing of the conditional signal. It is suggested that these cells may be involved in identifying a cue as relevant for initiation of a goal-directed movement. PMID- 9875569 TI - Two-photon laser-scanning microscopy: tests of objective lenses and Ca2+ probes. AB - The characteristics of objective lenses and Ca2+-sensitive probes were examined for imaging with a two-photon laser-scanning microscope (TP-LSM). The brightness of the images of beads taken by different objectives greatly varied and depended predominantly on their numerical aperture (NA) and less on transmittance and chirping effects. Lateral and axial resolutions, dx and dz, defined as the half decay length of fluorescence intensity of the image of a spherical bead (0.3 m) were 0.12 and 0.42 microm (objective; 40x/0.75). They are far better than those of confocal microscopes (0.3 and 1.5 microm, respectively) measured similarly (Kuba et al., 1994). dx linearly increased with an increase in 1/NA, while dz linearly increased with an increase in n/(NA)2 (n, refractive index) except for an objective of large NA (1.3). The coverslip compensation of objective lenses greatly affected the shape of the X-Z scanned images of 5.0 microm beads as well as resolutions, indicating a large effect of spherical aberration. Two-photon excitation spectra of Ca2+-sensitive fluorescent probes, indo-1, fura-2 and Oregon Green BAPTA-1, lied in a wavelength range shorter than twice that activated by one-photon absorption, while emission spectra were unchanged. Three dimensional images of a cultured hippocampal neurone loaded with Oregon Green BAPTA-1 showed fine structures of spines, dendrites and axons, while imaging with FM1-43 localized presynaptic boutons and demonstrated synaptic vesicle turnover. Dyes bleached little during the recording of 100 sectioned images. These characteristics of TP-LSM as well as its ability to image deeper tissues provide excellent means to study dynamic, spatial changes in intracellular substances and structures. To achieve the good performance of a TP-LSM, however, the relevant usage of appropriate objectives and fluorescent probes are required. PMID- 9875570 TI - Diversity of the T-cell receptor BV repertoire in HIV-1-infected patients reflects the biphasic CD4+ T-cell repopulation kinetics during highly active antiretroviral therapy. AB - OBJECTIVES: Highly active antiretroviral therapy (HAART) induces a decline in viral load and a biphasic increase in peripheral blood CD4+ T-cell counts in HIV infected patients. To evaluate the effect of HAART on T-cell receptor (TCR) diversity of repopulating naive and memory CD4+ T cells, complementarity determining region 3 (CDR3) spectratyping was performed. DESIGN: For four patients treated with HAART, CD45RO+ (memory) and CD45RA+ (naive) CD4+ T cells were isolated from peripheral blood leukocyte samples obtained 1 week before, 1-2 months after, and 9-11 months after start of treatment. METHODS: CDR3 regions were amplified by TCR-BV-specific nested PCR from CD4+ T-cell subsets. CDR3 size distributions and single-strand conformation polymorphism profiles were compared as an indication for TCR diversity. RESULTS: Increasing blood CD4+ T-cell counts during the first 2 months of treatment coincided with increased perturbation of CDR3 patterns in CD4+ T-cell subsets, suggesting an early oligoclonal repopulation. At later timepoints, CDR3 size diversity increased when T-cell counts did not substantially decrease. Memory and naive CD4+ T cells generally showed comparable levels of perturbation. CONCLUSION: Diversity of the TCR repertoire reflected biphasic T-cell repopulation during HAART, compatible with initial redistribution and later CD4+ T-cell production. Sustained elevation of T cell counts will in principle result in restoration of TCR diversity. PMID- 9875571 TI - Antiretroviral therapies in pregnancy: maternal, fetal and neonatal effects. Swiss HIV Cohort Study, the Swiss Collaborative HIV and Pregnancy Study, and the Swiss Neonatal HIV Study. AB - BACKGROUND: Therapies containing two reverse transcriptase inhibitors (RTI) with or without protease inhibitors are used with increasing frequency in pregnant HIV infected women. OBJECTIVE: To assess the safety of antiretroviral therapy in pregnant women and their newborns. METHODS: All clinical events and laboratory abnormalities in pregnant women on RTI with or without protease inhibitors and in their newborns were collected through an observational study. RESULTS: A total of 37 HIV-infected pregnant women have given birth to 30 children (by 30 April 1998). All received RTI, which were combined with protease inhibitors in 16 cases. Twelve women became pregnant while on treatment. Drugs used were as follows: zidovudine (n = 33), lamivudine (n = 33), stavudine (n = 4), indinavir (n = 9), ritonavir (n = 4), nelfinavir (n = 2) and saquinavir (n = 2). Adverse events during pregnancy were anaemia (n = 15), elevation of transaminases (n = 4), nausea/vomiting (n = 4), glucose intolerance (n = 2), nephrolithiasis (n = 2), diarrhoea (n = 2), hypertension (n = 1), insulin-requiring diabetes (n = 1). Adverse events in neonates were prematurity (n = 10), anaemia (n = 8), cutaneous angioma (n = 2), cryptorchidism (n = 2), transient hepatitis (n = 1). Non-life threatening intracerebral haemorrhage occurred in a premature baby (33 weeks gestation) exposed during fetal life to zidovudine-lamivudine-indinavir, and in a term baby exposed to stavudine-lamivudine-indinavir. Extrahepatic biliary atresia occurred in one newborn exposed to zidovudine-lamivudine-indinavir. Maternal viral load was below 400 copies/ml in 18 out of 30 patients who delivered. One case of mother-to-child HIV transmission was identified. CONCLUSIONS: In HIV infected pregnant women treated with two RTI with or without protease inhibitors, one or more adverse events occurred in 29 out of 37 women and in 14 out of 30 babies. In newborns, frequent prematurity, one case of biliary malformation and one intracerebral haemorrhage in a term baby are of concern. These observations do not preclude combination therapies during pregnancy but emphasize the necessity to maintain updated registers on their safety. PMID- 9875572 TI - Changes in renal function associated with indinavir. AB - BACKGROUND: Indinavir use is associated with a spectrum of renal and urinary tract complications including nephrolithiasis, renal colic and pain without recognizable lithiasis, and a picture of crystalluria-dysuria. A frank nephropathy has not been recognized as part of the spectrum. METHODS: A retrospective analysis of 106 HIV-infected individuals receiving indinavir was performed with the purpose of identifying the frequency and risk factors for indinavir-associated nephropathy and urinary complications. Individuals receiving ritonavir or nelfinavir served as controls. RESULTS: A sustained elevation of creatinine (>20%, into abnormal range) was identified in 20 (18.6%) subjects treated with indinavir but not with other protease inhibitors. Creatinine elevation was associated with treatment duration of more than 54 weeks [odds ratio (OR), 7.1; 95% confidence interval (CI), 1.8-27.7], low baseline body mass index < or = 20 kg/m2 (OR, 4.0; 95% CI, 1.0-16.6), and use of trimethoprim sulphamethoxazole (TMP-SMX; OR, 4.6; 95% CI, 1.5-13.8). Lower urinary specific gravity (P = 0.015), and leukocyturia (P<0.001) were frequently associated features of indinavir nephropathy. No patient developed severe renal impairment and abnormalities were reversible upon discontinuation of the drug. Complications (renal colic, or pain and dysuria) occurred after a mean of 36 weeks (95% CI, 23 48) of indinavir treatment in 13 subjects (12.3%), eight of whom (62%) presented elevated creatinine during follow-up. Only long-term exposure to TMP-SMX (>160 weeks) was identified as a potential risk for the occurrence of a clinical event (OR, 4.7; 95% CI, 1.2-19.2). CONCLUSIONS: A crystal nephropathy, characterized by serum creatinine elevation, loss of concentrating ability of the kidney, leukocyturia, and renal parenchymal image abnormalities, is a frequent complication of indinavir therapy. Identification of individuals at risk, particularly those with low body mass index or receiving TMP-SMX prophylaxis, may help the decision to initiate indinavir or chose an alternative protease inhibitor in order to minimize renal and urinary tract adverse events. PMID- 9875573 TI - Lest we forget: neuropsychiatry and the new generation anti-HIV drugs. PMID- 9875574 TI - Variations in HIV-1 pol gene associated with reduced sensitivity to antiretroviral drugs in treatment-naive patients. AB - OBJECTIVE: Various drugs against the HIV-1 enzymes reverse transcriptase (RT) and protease have been introduced in the last few years: protease inhibitors, nucleoside RT inhibitors (NRTI) and non-NRTI (NNRTI). Several sequence variations associated with reduced drug sensitivity have been described in the HIV-1 pol gene. DESIGN: To analyse the occurrence of mutations associated with drug resistance in treatment-naive individuals. METHODS: RNA was extracted from sera of treatment-naive individuals, who were first diagnosed to be HIV-1 infected between August 1996 and February 1998. The pol region was amplified by RT-PCR and directly sequenced. Data on mutations associated with resistance to antiretroviral drugs were obtained from literature. RESULTS: Fifty protease genes and 53 RT genes from 57 individuals were sequenced. In the RT we analysed 20 amino-acid positions associated with resistance to NRTI and NNRTI. In total, 1054 amino acids at critical positions were analysed and three (0.3%) mutations known to contribute to RTI resistance were detected. In the protease, 16 amino-acid positions associated with resistance to protease inhibitors were analysed. By analysing a total of 768 amino acids at key positions in the protease, 50 (7%) mutations were detected that were associated with reduced drug sensitivity. Thirty-one (61%) patients showed between one and six mutations at the analysed protease amino-acid positions. In eight out of 16 analysed amino-acid positions, up to 44% of all patients carried mutations associated with resistance to protease inhibitors. CONCLUSIONS: Very few pre-existing mutations to RTI were found, suggesting that the transmission of RT-resistant strains is still uncommon. However, about two-thirds of the patients had one or more mutations associated with resistance to protease inhibitors. In addition, at some amino acid positions up to almost half of the patients carried variations claimed to contribute to protease inhibitor resistance. Most of these mutations are likely to reflect the natural polymorphism of the protease. Their impact on the long term effect of antiretroviral treatment should be evaluated in future studies. PMID- 9875575 TI - HIV-1 gp120 increases the permeability of rat brain endothelium cultures by a mechanism involving substance P. AB - OBJECTIVE: To analyse whether an HIV-1 envelope protein might play a role in damaging the blood-brain barrier as a fundamental step in the early invasion of the central nervous system by HIV-1. DESIGN: Analysis of permeability of rat brain endothelium cultures to albumin, to assess the functional integrity of the vascular component of the blood-brain barrier. METHODS: Rat brain endothelium cultures prepared by cerebral microvessels were exposed to recombinant gp120IIIB on microporous membranes and passage of biotin-labelled albumin was analysed. Scanning electron microscopy was used to analyse cell culture morphology. Some cultures were preincubated with N-nitro-L-arginine methyl ester (L-NAME), a selective inhibitor of nitric oxide synthase, or with spantide, a selective substance P antagonist. RESULTS: HIV-1 gp120 increased the permeability of rat brain endothelial cells to albumin in a dose-dependent manner. Scanning electron microscopy revealed profound gp120-induced alterations in cell morphology accounting for the increased permeability to macromolecules. These alterations were neutralized by anti-gp120 monoclonal antibody but not by isotype control antibody or L-NAME. By contrast, spantide and anti-substance P polyclonal antibody completely blocked the gp120-induced increase in albumin permeability. Control cultures exposed to measles virus nucleoprotein showed an increase in permeability that was not blocked by spantide. Brain endothelial cells, exposed to gp120, displayed cell surface immunoreactivity for substance P, suggesting that substance P is secreted by brain endothelium in response to gp120 stimulation and binds to brain endothelial cells through a receptor-mediated mechanism. CONCLUSIONS: These findings suggest a role for substance P in the gp120-induced increase in permeability of brain endothelium. PMID- 9875577 TI - Protease inhibitor and triple-drug therapy: cellular immune parameters are not restored in pediatric AIDS patients after 6 months of treatment. AB - OBJECTIVE: To assess whether treatment of HIV-positive children by antiretroviral drugs for a 6-month period would improve immune function significantly. DESIGN AND METHODS: Immunological assessment of 89 HIV-positive children who received protease inhibitor monotherapy for 12-16 weeks as part of phase I/II studies, followed by triple antiretroviral therapy for an additional 12 weeks, was conducted. Immunological parameters were assessed in vitro at four time points (at enrollment, at weeks 2-4, at weeks 12-16, and at weeks 24-28). Assessments included: cytokine production by monocytes, T-cell proliferation to mitogen or recall antigens (including an HIV antigen) and apoptotic cell death. Plasma levels of tumor necrosis factor (TNF)-alpha and soluble TNF receptor (sTNF-R) were also measured, in addition to CD4+ T-lymphocyte counts and viral load. In addition, limited analyses were performed on samples from 17 children after 120 weeks of therapy, including 104 weeks of triple therapy. RESULTS: At enrollment, the 89 children exhibited severe immune defects. Antiretroviral therapy raised CD4+ T-lymphocyte counts significantly and decreased viral loads. In contrast, the in vitro immune parameters studied were not improved, except for plasma levels of sTNF-RII which decreased in parallel with the decrease in viral load. In addition, there was a trend towards increased skin test reactivity for the ritonavir-treated children. No differences were seen in the immune parameters whether the patients were treated with mono- or triple therapy. Results obtained after 120 weeks of therapy demonstrated that defective interleukin-12 production was not restored by long-term therapy. CONCLUSIONS: After 6 months of therapy, with the exception of decreased sTNF-RII levels, and a trend towards increased skin test reactivity, restoration of several defective cellular immune responses did not occur despite significantly decreased viral loads and increased CD4+ T lymphocyte counts. PMID- 9875576 TI - Immune activation in HIV-infected African individuals. Italian-Ugandan AIDS cooperation program. AB - OBJECTIVE: Immune activation induced by chronic infections, dietary limitations, and poor hygienic conditions is suggested to be present in African HIV infection and is at the basis of the hypothesis that HIV infection in Africa could be prevalently associated with immunopathogenetic mechanisms. Very limited data are nevertheless available supporting this theory, and in particular no data are reported on functional and phenotypic analyses performed on fresh peripheral blood mononuclear cells (PBMC) of African HIV-infected patients living in Africa. DESIGN: Immunological and virological parameters were analysed in fresh PBMC of HIV-infected African and Italian patients with advanced HIV disease and comparable CD4 and CD8 counts, sex, and age. Both functional (antigen- and mitogen-stimulated cytokine production) and phenotypic (activation markers; markers preferentially expressed by T helper (Th) type 2 cells or by memory and naive cells) analyses were performed. Results were compared with those of HIV seronegative African and Italian controls. HIV plasma viraemia was analysed by competitive polymerase chain reaction (PCR) and branched DNA techniques. RESULTS: (1) The production of mitogen-stimulated IFN-gamma and TNF-alpha as well as the production of env peptide-stimulated IFN-gamma, TNF-alpha, and IL-10 are increased in African HIV infection; (2) the expression of activation and Th2 associated markers is augmented in African HIV infection as is the memory/naive ratio; (3) mitogen-stimulated IFN-gamma and IL-10 production, as well as the expression of activation and Th2-associated markers and the memory/naive ratio, are augmented in African compared with Italian controls; and (4) plasma viraemia is reduced in African compared with Italian HIV-infected individuals. CONCLUSIONS: These results, which are the first to be reported on fresh material from African HIV-infected patients living in Africa, indicate that HIV disease is associated with an abnormal immune hyperactivation and may be accompanied in these patients by lower loads of virus, and show that such activation is present even in HIV-seronegative controls. PMID- 9875579 TI - Evaluation of guidelines for initiation of highly active antiretroviral therapy in a longitudinal cohort of HIV-infected individuals. AB - OBJECTIVES: Expert panels have developed several guidelines for initiating highly active antiretroviral therapy (HAART) in patients with HIV infection. To evaluate these guidelines, we simulated their application in a cohort of HIV-infected patients established and followed before HAART was available, and determined how long such patients survived without disease progression in the absence of HAART. METHODS: Longitudinal data was used that had been collected from 1982 to 1995 on a prospective cohort of 133 homosexual men with known or closely approximated dates of HIV-1 seroconversion and negligible antiretroviral exposure. The main definition of disease progression was CD4 cell count < or = 300x10(6)/l or development of clinical AIDS diagnosis within 12 months. RESULTS: The mean number of years between the recommended initiation of therapy and when disease progression occurred in the absence of HAART were as follows: initiation of treatment at first visit, 4.81 years [median, 3.78 years; interquartile range (IQR), 1.85-6.59 years]; CD4 cell count <500x10(6)/l or serum RNA >5000 copies/ml (at least 10000 copies/ml fresh plasma), 4.35 years (median, 3.22 years; IQR, 1.56-6.19 years); CD4 cells <500x10(6)/l or serum RNA >20000 copies/ml (at least 40000 copies/ml fresh plasma), 3.61 years (median, 2.70 years; IQR, 1.40-5.11 years); and CD4 cells <500x10(6)/l, 2.72 years (median, 2.17 years; IQR, 0.81 4.25 years). The percentage of patients who had disease progression before HAART would have been recommended was 0.8, 1.6, 3.2 and 13.6% with each of these four approaches, respectively. CONCLUSIONS: Implementation of recommended treatment guidelines will result in a substantial proportion of patients being treated for long periods before immunologic or clinical disease progression would have occurred in the absence of HAART. These findings should be considered in the clinical care of HIV-infected patients and in future recommendations for the initiation of HAART. PMID- 9875578 TI - Induction of immune responses to HIV-1 by canarypox virus (ALVAC) HIV-1 and gp120 SF-2 recombinant vaccines in uninfected volunteers. NIAID AIDS Vaccine Evaluation Group. AB - OBJECTIVE: To determine the ability of live attenuated canarypox virus expressing HIV antigens to induce CD8+ cytotoxic T-cell responses and to prime for neutralizing antibody responses to boosting with purified recombinant gp120 subunit vaccine. DESIGN: A prospective, double-blind, randomized, immunogenicity and safety study was conducted in healthy adults at low risk for acquiring HIV infection and who were seronegative for HIV. METHODS: CD8+ cytotoxic T-cells directed against Env or Gag expressing target cells were measured after live recombinant canarypox-HIV-1 vaccine priming (vaccine given at days 0, 7, 14 and 21). Neutralizing antibodies were measured after subunit boosting (vaccine given at days 28 and 84). RESULTS: CD8+ CTL were induced in 64% of volunteers by the live recombinant canarypox-HIV-1 vaccine. All volunteers who received two doses of subunit vaccine after live recombinant canarypox priming developed neutralizing antibodies directed against laboratory strains of HIV-1 and seven out of eight volunteers tested developed neutralizing antibodies to the primary isolate, BZ167, but to none of eight other primary isolates. Unprimed controls had low or absent neutralizing antibodies after two doses of subunit vaccine. CONCLUSIONS: The live canarypox vector was safe, stimulated cytotoxic T-cells and primed for a vigorous neutralizing antibody response upon boosting with subunit gp120 vaccine. This vaccine combination should be evaluated further for inducing protection against HIV infection. PMID- 9875580 TI - Neuromuscular function in HIV infection: analysis of a placebo-controlled combination antiretroviral trial. AIDS Clinical Group 175/801 Study Team. AB - OBJECTIVE: To determine the frequency of peripheral neuropathy and myopathy in HIV-infected subjects enrolled in a combination antiretroviral treatment trial. DESIGN AND METHODS: AIDS Clinical Trial Group (ACTG) protocol 175 was a multicenter, double-blind, placebo-controlled, clinical trial. A total of 2467 subjects were randomized to one of four single or combination regimens, containing zidovudine (ZDV), didanosine (ddl), zalcitabine (ddC), and their respective placebos. Site investigators reported peripheral neuropathy, and the diagnosis of distal symmetrical neuropathy (DSP) was established by the study authors. Myalgia, muscle weakness and creatine phosphokinase (CPK) were prospectively assessed in a subset of the antiretroviral-naive cohort (n = 1067). RESULTS: Of 222 site diagnoses of neuropathy, 109 (49%) were DSP. There was a significant difference between treatment arms for rate of DSP and time to first grade 2 or higher DSP (ZDV-ddC, 6%; ZDV, 4%; ZDV-ddl, 4%; ddl, 3%; P = 0.029). Age and Karnofsky score were significant predictors of DSP. Fifty-six (54%) out of 104 patients with DSP remained on study medication at full (n = 29) or reduced (n = 27) dose within 6 months of developing neuropathy. There was no significant difference between treatment arms in the rate of myalgia or muscle weakness. The median CPK of subjects on ZDV-ddC was significantly higher than other study treatments, although CPK levels did not correlate with symptoms of myopathy. Only six subjects were diagnosed with myopathy during the study (one ZDV-ddl, one ZDV ddC, and four ddl). CONCLUSIONS: DSP and myopathy may occur with current dosing regimens of combination antiretroviral therapy, and should be diagnosed using stringent criteria. ZDV-ddC was associated with the highest rate of DSP, although features of myopathy were not significantly different between treatment regimens. PMID- 9875581 TI - Variation in incidence of indinavir-associated nephrolithiasis among HIV-positive patients. AB - BACKGROUND: Nephrolithiasis may be an important consequence of indinavir therapy; however little has been published on the variation in incidence between different populations of patients or the possible mechanisms of calculus formation. OBJECTIVE: To examine variation in the incidence of indinavir-associated nephrolithiasis (IAN) in HIV-positive patients in relation to hemophilia and hepatitis C virus (HCV) infection. METHODS: Clinical data were abstracted retrospectively from the medical records of all adult patients treated with indinavir from September 1995 to September 1997. Occurrence of first IAN, defined as flank pain and hematuria after initiation of therapy, was analyzed in relation to hemophilia status and HCV infection. RESULTS: There were 17 episodes of IAN (22%) among 79 patients treated with indinavir. Of 10 patients with hemophilia, 50% developed IAN as compared with 17% of patients without hemophilia (P = 0.03). Median days to first IAN was 22 (range 7-110 days) for hemophiliacs and 156 (range 5-611 days) for those without hemophilia. Data for HCV status were available for 74 out of 79 patients: 10 out of 27 (37%) patients with HCV developed IAN compared with six out of 42 (14%) without HCV (P = 0.02). CONCLUSION: Overall incidence of IAN was higher than that previously reported and was significantly greater in hemophiliacs than in non-hemophiliacs. HCV may be a contributing factor. PMID- 9875582 TI - A phase II/III trial of antimicrobial therapy with or without amikacin in the treatment of disseminated Mycobacterium avium infection in HIV-infected individuals. AIDS Clinical Trials Group Protocol 135 Study Team. AB - OBJECTIVE: To determine the clinical and microbiologic benefit of adding amikacin to a four-drug oral regimen for treatment of disseminated Mycobacterium avium infection in HIV-infected patients. DESIGN: A randomized, open-labeled, comparative trial. SETTING: Outpatient clinics. PATIENTS: Seventy-four patients with HIV and symptomatic bacteremic M. avium infection. INTERVENTIONS: Rifampin 10 mg/kg daily, ciprofloxacin 500 mg twice daily, clofazimine 100 mg every day, and ethambutol 15 mg/kg orally daily for 24 weeks, with or without amikacin 10 mg/kg intravenously or intramuscularly 5 days weekly for the first 4 weeks. MAIN OUTCOME MEASURE: Clinical and microbiologic response at 4 weeks; quantitative level of bacteremia with M. avium. RESULTS: No difference in clinical response was noted with the addition of amikacin to the four-drug oral regimen, and only 25% in either group had a complete or partial response at 4 weeks. A comparable quantitative decrease in bacteremia was noted in both treatment groups, with 16% of patients being culture-negative at 4 weeks and 38% at 12 weeks. Toxicities were mainly gastrointestinal. Amikacin was well tolerated. Median survival was 30 weeks in both groups. CONCLUSIONS: The addition of amikacin to a four-drug oral regimen of rifampin, ciprofloxacin, clofazimine, and ethambutol did not provide clinical or microbiologic benefit. PMID- 9875583 TI - Twice weekly tuberculosis preventive therapy in HIV infection in Zambia. AB - BACKGROUND: A randomized double-blind placebo-controlled trial was conducted to estimate the efficacy of preventive therapy for tuberculosis (TB) in HIV-infected adults in Lusaka, Zambia. The main outcome measures were the incidence of TB, mortality and adverse drug reactions. METHODS: During a 2 year period, 1053 HIV positive individuals without evidence of clinical TB were randomly assigned to receive 6 months of isoniazid twice a week (H), or 3 months of rifampicin twice a week (R) plus pyrazinamide (Z), or a placebo. Therapy was taken twice a week and was self administered. Subjects presenting with symptoms during the follow-up period were investigated for TB. RESULTS: The 1053 subjects in the study were followed up for a total of 1631 person-years (median = 1.8 years). Twenty-nine subjects were taken off treatment as a result of adverse drug reactions. A total of 96 cases of TB/probable TB (59 TB and 37 probable TB) were diagnosed during the study period and 185 deaths were reported. One hundred and fifteen subjects (11%) did not return to the study clinic at any time after enrolment. The incidence of TB was lower in those subjects on preventive therapy (H and RZ groups combined) compared with those on placebo (rate ratio = 0.60, 95% CI: 0.36 1.01, P = 0.057), as was the incidence of TB/probable TB (rate ratio = 0.60, 95% CI: 0.40-0.89, P = 0.013). The effect of preventive therapy was greater in those with a tuberculin skin test (TST) of 5 mm or greater, in those with a lymphocyte count of 2x10(9)/l or higher, and in those with haemoglobin of 10 g/dl or higher. There was no difference in mortality rates between the preventive therapy and placebo groups. The effect of preventive therapy declined after the first year of the study so that by 18 months the rates of TB in the treated groups were similar to that in the placebo group. CONCLUSION: This study has demonstrated that preventive therapy with either twice weekly isoniazid for 6 months or a combination of rifampicin and pyrazinamide for 3 months reduced the incidence of TB in HIV-infected persons in Zambia. No effect was observed on mortality. The effect was greatest in persons who had a positive TST or a lymphocyte count of 2x10(9)/l or greater, indicating that preventive therapy may be more effective in people with less advanced immunosuppression. The limited duration of the protective effect reported in this study raises the question of the need for lifelong preventive therapy or re-prophylaxis. PMID- 9875584 TI - A clinicopathologic analysis of AIDS-related cryptosporidiosis. AB - OBJECTIVE: To characterize the histology of AIDS-associated cryptosporidiosis and identify features that explain the clinical variability. DESIGN: A retrospective analysis of HIV-positive individuals with cryptosporidiosis who underwent endoscopy at the Johns Hopkins Hospital between 1985 and 1996. METHODS: The histologic features (intensity of Cryptosporidium infection, inflammation, mucosal damage, copathogens) of gastrointestinal biopsies from 37 HIV-positive individuals with cryptosporidiosis were systematically graded. These histologic features were correlated with the severity of the diarrheal illness obtained from a patient chart review. RESULTS: Histologic features associated with Cryptosporidium infection include a neutrophilic infiltrate in the stomach, villus blunting in the duodenum, cryptitis and epithelial apoptosis in the colon, and reactive epithelial changes in the stomach and duodenum. The nature and intensity of the inflammatory response varied widely; however, duodenal biopsies from a subset of patients (37%) revealed marked acute inflammation that was associated with concomitant cytomegalovirus infection. Although duodenal infection was common (93% of individuals), infection of other sites was variable (gastric cryptosporidiosis in 40% and colonic cryptosporidiosis in 74%). Widespread infection of the intestinal tract, which included both the large and small intestine, was associated with the most severe diarrheal illness. CONCLUSIONS: Cryptosporidium infection produces histologic evidence of gastrointestinal mucosal injury. The inflammatory response to the infection is variable, and may be modified by copathogens such as cytomegalovirus. The clinical manifestations are influenced, in part, by the anatomic distribution of the infection, with extensive infections involving both small and large intestines producing the most severe illness. PMID- 9875585 TI - Treatment of AIDS-associated progressive multifocal leukoencephalopathy with highly active antiretroviral therapy. AB - OBJECTIVES: To evaluate the efficacy of highly active antiretroviral therapy (HAART) in 12 patients with AIDS-associated progressive multifocal leukoencephalopathy (PML). PATIENTS AND METHODS: The diagnosis of PML was established by brain biopsy in six patients and by neuroimaging findings and PCR detection of JC virus in cerebrospinal fluid (CSF) in six patients. We also studied 13 consecutive AIDS patients with biopsy-proven PML cared for in the same institution before HAART was available. Eleven patients of the HAART group and eight patients of the control group received intravenous arabinoside cytosine cycles. RESULTS: With HAART, the median decrease in the HIV viral load was 3.58 log10 copies/ml and the median increase in the CD4 cell count was 74x10(6)/l. The median survival time after PML diagnosis was 545 days in the HAART group and 60 days in the control group (P<0.001, log-rank test). In the HAART group, the neurological deficits improved substantially in six patients and stabilized in six patients. Eleven patients underwent follow-up cranial computed tomography or magnetic resonance scan that showed improvement of PML lesions in 10 patients and stabilization in one patient. Follow-up CSF analysis showed clearance of JC virus in six out of seven patients who had an initial positive result. CONCLUSIONS: This study shows that HAART may increase the survival, clinical status and radiological features of AIDS patients with PML. Clearance of JC virus from CSF has been found, suggesting that immune reconstitution can interrupt the JC virus lytic cycle. PMID- 9875586 TI - Increased protected sex and abstinence among Namibian youth following a HIV risk reduction intervention: a randomized, longitudinal study. AB - OBJECTIVE: To evaluate an HIV risk-reduction intervention among Namibian adolescents. METHODS: A randomized trial of a 14-session face-to-face intervention emphasizing abstinence and safer sexual practices conducted among 515 youths (median age 17 years and median grade 11) attending 10 secondary schools located in two districts in Namibia. Youths were randomly assigned to the intervention or control condition at the level of the individual. HIV risk behaviours, intentions and perceptions were assessed at baseline, immediately post-intervention and at 6 and 12 months post-intervention. RESULTS: Among all 515 youths who enrolled in the programme, rates of either abstinence or sex with a condom were not different between control and intervention youths at baseline or in the follow-up period. However, analyses conducted among the subset of youths who were sexually inexperienced at baseline (n = 255) revealed that a higher percentage of intervention youths (17%) than control youths (9%, P<0.05) remained sexually inexperienced one year later. Moreover, in the immediate post intervention period, among baseline virgins who subsequently initiated sex, intervention youths were more likely than control youths to use a condom (18 versus 10%, P<0.05). Additional HIV-related risk behaviours (failure to discuss previous HIV risk exposure with one's sexual partner and alcohol use), intentions to use condoms, and perceptions of the ability to use condoms were positively affected by the intervention. CONCLUSIONS: There is evidence that the 'My Future is My Choice' (MFMC) intervention is reducing HIV risk behaviours among sexually inexperienced participants aged 15-18. Related risk behaviours and perceptions are also positively impacted by the intervention. PMID- 9875587 TI - Seroconversion for human herpesvirus 8 during HIV infection is highly predictive of Kaposi's sarcoma. AB - BACKGROUND: The finding of antibodies against human herpesvirus 8 (HHV-8) is associated with the occurrence of Kaposi's sarcoma in persons infected with HIV. However, the predictive value of HHV-8 antibodies for Kaposi's sarcoma in HIV infection is unknown. METHODS: The Amsterdam Cohort Studies on HIV infection and AIDS started in 1984 for homosexual men and in 1985 for injecting drug users. Serum samples from 1459 homosexual men and 1167 drug users were tested for antibodies to recombinant HHV-8 lytic-phase capsid (ORF65) antigen and latent phase nuclear (ORF73) antigen. Individuals were retrospectively identified as HHV 8-positive or HHV-8-negative at enrolment or HHV-8 seroconverter during the study. Kaposi's sarcoma-free survival time was compared between HIV-infected men who were positive for HHV-8 at enrolment and those who later seroconverted for HHV-8. Hazard ratios were estimated for Kaposi's sarcoma, lymphoma, and opportunistic infection according to the HHV-8 serostatus. RESULTS: The incidence of HHV-8 seroconversion among drugs users was 0.7 per 100 person-years based on 31 seroconversions, whereas an incidence of 3.6 was found among homosexual men based on 215 seroconversions. The hazard ratio for Kaposi's sarcoma was 3.15 (95% CI: 1.89-5.25) in HIV-infected individuals if HHV-8 antibodies were present either at enrolment or at HIV seroconversion. In HIV-infected persons who later seroconverted to HHV-8, Kaposi's sarcoma developed more rapidly: hazard ratio of 5.04 (95% CI: 2.94-8.64), an additional risk of 1.60 (95% CI: 1.01-2.53; P = 0.04). Time-dependent adjustment for CD4+ cell count and HIV RNA had no impact on the additional risk, although the CD4+ cell count was an independent risk factor for Kaposi's sarcoma. HHV-8 infection did not increase the risk of AIDS-related lymphoma or opportunistic infections. CONCLUSIONS: The incidence of HHV-8 infection is higher in homosexual men than in drug users. The presence of HHV-8 antibodies in HIV-infected persons increases the risk of Kaposi's sarcoma. Among HIV-infected persons, those who subsequently seroconvert for HHV-8 are at highest risk. These results strongly confirm the causal role of HHV-8 in Kaposi's sarcoma and emphasize the clinical relevance of HHV-8 seroconversion before and after the HIV infection. PMID- 9875589 TI - Combination antiretroviral therapy during pregnancy. PMID- 9875588 TI - Acceptability of voluntary HIV testing by pregnant women in developing countries: an international survey. Ghent International Working Group on Mother-to-Child Transmission of HIV. AB - OBJECTIVE: To evaluate acceptability of voluntary HIV counselling and testing (VCT) by pregnant women in the context of clinical trials assessing interventions to reduce mother-to-child transmission (MCT) of HIV in developing countries. METHODS: During September-October 1997, 13 studies located in West (Abidjan, Bobo Dioulasso), East (Nairobi, Mombasa, Dar Es Salaam, Blantyre, Lusaka, Harare) and South Africa (Soweto, Durban), and Thailand (Bangkok) were included in a cross sectional mailing survey about the acceptability of VCT in antenatal clinics. Acceptance rate, return rate, overall acceptability of VCT (acceptance of both pre- and post-VCT sessions) were obtained using a standardized questionnaire. RESULTS: The median overall acceptability of VCT was 69% (range, 33-95%). Overall acceptability of VCT most frequently depended on return rates because acceptance rates were generally high. Where several studies were conducted in parallel in the same city or the same country, overall acceptability rates of HIV testing were generally comparable even if the intervention programmes differed. Overall acceptability rates of VCT were high in antenatal clinics where a particular effort in implementing VCT programmes had been made. CONCLUSIONS: This international survey shows that despite many obstacles, VCT is feasible and acceptable for pregnant women aiming to reduce their risk of transmitting HIV to their children. PMID- 9875590 TI - Severe coronary artery disease in a young HIV-infected man with no cardiovascular risk factor who was treated with indinavir. PMID- 9875591 TI - Early increase in cell-associated HIV-1 DNA in patients on highly active antiretroviral therapy. PMID- 9875592 TI - Survival of HIV-infected patients with cytomegalovirus retinitis under highly active antiretroviral therapy. Groupe d'Epidemiologie Clinique du SIDA en Aquitaine. PMID- 9875593 TI - Diagnosis of HIV-1/2 dual infection using dilution analysis of type-specific antibody. PMID- 9875595 TI - HIV serostatus of sexual partners of HIV-positive and HIV-negative homosexual men in Sydney. PMID- 9875594 TI - Immunologic and virologic response of HIV-2 infection to antiretroviral therapy. PMID- 9875596 TI - Testing for HIV in California: volume and seropositivity in private versus public sites. PMID- 9875597 TI - Therapeutic drug monitoring of methadone in HIV-infected patients receiving protease inhibitors. PMID- 9875599 TI - Withdrawal of fluconazole suppressive therapy for thrush in patients responding to combination antiviral therapy including protease inhibitors. PMID- 9875598 TI - Effect of highly active antiretroviral therapy on recurrent oropharyngeal candidiasis in HIV-infected patients. PMID- 9875600 TI - Investigation of multiaxial molecular dynamics by 2H MAS NMR spectroscopy. AB - The technique of 2H MAS NMR spectroscopy is presented for the investigation of multiaxial molecular dynamics. To evaluate the effects of discrete random reorientation a Lie algebraic formalism based on the stochastic Liouville-von Neumann equation is developed. The solution to the stochastic Liouville-von Neumann equation is obtained both in the presence and absence of rf irradiation. This allows effects of molecular dynamics to be evaluated during rf pulses and extends the applicability of the formalism to arbitrary multiple pulse experiments. Theoretical methods are presented for the description of multiaxial dynamics with particular emphasis on the application of vector parameters to represent molecular rotations. Numerical time and powder integration algorithms are presented that are both efficient and easy to implement computationally. The applicability of 2H MAS NMR spectroscopy for investigating molecular dynamics is evaluated from theoretical spectra. To demonstrate the potential of the technique the dynamics of thiourea-2H4 is investigated experimentally. From a series of variable temperature MAS and quadrupole echo spectra it has been found that the dynamics can be described by composite rotation about the CS and CN bonds. Both experiments are sensitive to the fast CS rotation which is shown to be described by the Arrhenius parameters E(CS) = 46.4 +/- 2.3 kJ mol(-1) and ln(A(CS))= 32.6 +/- 0.9. The MAS experiment represents a significant improvement by simultaneously allowing the dynamics of the slow CN rotation to be fully characterized in terms of E(CN) = 56.3 +/- 3.4 kJ mol(-1) and ln(A(CN)) = 25.3 +/ 1.1. PMID- 9875601 TI - Molecular motion in solid all-trans retinoic acid (vitamin A acid) by proton NMR. AB - An investigation of the molecular dynamics of all-trans retinoic acid (vitamin A) has been carried out in the solid state by proton NMR. Measurements were made of T1 at 14 and 25 MHz and of second moment at 25 MHz in the temperature range 55 to 420 K for both monoclinic and triclinic forms. In the monoclinic state relaxation was attributed to methyl group reorientation below 250 K and to conformational motion of the cyclohexene ring above this temperature. Activation energies and time constants were derived for the motions. In the triclinic state the same motions were active, but the cyclohexene ring motion was less restricted and some of the methyl group motions were more restricted indicating that hindrances to the motions were at least partly determined by the intermolecular packing arrangements. PMID- 9875602 TI - Magnitudes and mutual orientations of dipolar and shielding interaction tensors determined from the orientation dependence of spinning sidebands of slowly rotating powder samples. AB - It is shown that the orientation dependence of spinning sidebands of slowly rotating solids can be exploited to determine with good precision the heteronuclear bond distances and mutual orientation of dipolar and shielding tensors from a single dipolar modulated chemical shift anisotropy spectrum. The method is based on the dynamics of polarization transfer in the initial period of tens of microseconds, and may be useful in solids with relatively isolated groups of spins. PMID- 9875603 TI - Calculation of the NMR second moment for materials with different types of internal rotation. AB - The measurements of the NMR second moment can reveal information about structural and dynamical details of the sample providing that analysis of the experimental results on the basis of the Van Vleck's formula can be performed. In this paper the method of calculating the Van Vleck's second moment for solids with complex internal motions is presented. The method is based on simulating any desired motion of atoms within a block of unit cells large enough to reflect the property of the macroscopic sample and calculating the second moment value averaged by this motion. The detailed description of this method is given for the case of rotation of molecules or groups of atoms. The algorithm of the computer program which performs calculation based on the described method is also presented. Examples of application of the described method are listed. PMID- 9875604 TI - 13C CP (cross-polarization) MAS (magic angle spinning) NMR and GIAO-CHF calculations of buspirone analogues. Part 1. 3a,4,7,7a-Tetrahydro-2-[4-[4-(2 quinolinyl)-1-piperazinyl]butyl ]-4,7-ethane-1H-isoindole-1,3(2H)-dione hydrochloride and hydrobromide. AB - 13C CP (cross-polarization) MAS (magic angle spinning) solid state NMR spectra of buspirone analogue 3a,4,7,7a-tetrahydro-2-[4-[4-(2-quinolinyl)-1 piperazinyl]butyl]-4,7-eth ane-1H-isoindole-1,3(2H)-dione were recorded. In the spectra of hydrochloride and hydrobromide, two sets of signals appeared, in agreement with single crystal X-ray diffraction data indicating that in each of the salts two independent cations were present in the crystal unit. The largest shielding differences of 3.2-4.6 ppm between two sets of signals were found for quinoline aromatic carbons C3 and C2. Ab initio calculations of the carbon and nitrogen shielding constants were performed with the use of the GIAO-CHF method for structural fragments: N-butylsuccinimide, quinoline-(N-methyl) piperazine hydrochloride and hydrobromide. Linear correlations between theoretical and solid state results were obtained, thus enabling a reasonable assignment of carbon resonances of the conformations present in the solid state. Due to the fast dynamics in solution, the carbon chemical shifts corresponded to the averaged values of the forms present in the solid state. PMID- 9875605 TI - Anisotropy of chemical shift and J coupling for P-31 and Se-77 in trimethyl and triphenyl phosphine selenides. AB - The 31P and 77Se magic angle spinning (MAS) nuclear magnetic resonance (NMR) experiments for selenium-77 enriched (70%) trimethylphosphine selenide 1 and triphenylphosphine selenide 2 were carried out in order to determine the nuclear magnetic shielding tensors of both nuclei and to establish values of the phosphorus-selenium indirect spin-spin coupling anisotropy delta J. The m = +1/2 and m = -1/2 subspectra were analysed by the dipolar-splitting-ratio method of Eichele and Wasylischen. For the C(S) molecule 1, delta J was obtained to be +640 +/- 260 Hz from the 31P spectrum and +550 +/- 140 Hz from the 77Se spectrum. Density functional theory (DFT) calculations give a delta J value of about +705 Hz. The value of delta J could not be determined unambiguously by analysis of the 31P spectra for the C1 molecules 2; nevertheless, an estimation of delta J was possible. The principal axis 3 of the phosphorus shielding tensor was determined to be nearly parallel to the PSe bond in 1 and 2. For the selenium shielding of 1, the same orientation was found, whereas in 2, the principal axis 2 of the selenium shielding was found to be oriented nearly along the PSe bond. The experimentally determined phosphorus nuclear magnetic shielding tensors agree well with those calculated by the IGLO method. For those two principal values of the selenium-shielding tensors corresponding to directions nearly perpendicular to the SeP bond, the agreement between calculated and experimental values is satisfactory. For the third one, corresponding to the principal axis close to the SeP bond, the calculated deshielding contributions are distinctly too small for both compounds investigated. Trends observed for the calculated molecular orbital (MO) contributions to the shielding as well as possible reasons for the underestimation of the deshielding contributions along the SeP bond are discussed. PMID- 9875606 TI - A comparison of double rotation and multi-quantum magic angle spinning spectra. AB - A field and method independent canonical representation of high-resolution spectra of quadrupole nuclei enables universal analyses of experimental data. The best accuracy from 1D measurements is achieved if DOR and MQ-MAS methods are combined. PMID- 9875607 TI - The influence of shrinkage-cracking on the drying behaviour of White Portland cement using Single-Point Imaging (SPI). AB - The removal of water from pores in hardened cement paste smaller than 50 nm results in cracking of the cement matrix due to the tensile stresses induced by drying shrinkage. Cracks in the matrix fundamentally alter the permeability of the material, and therefore directly affect the drying behaviour. Using Single Point Imaging (SPI), we obtain one-dimensional moisture profiles of hydrated White Portland cement cylinders as a function of drying time. The drying behaviour of White Portland cement, is distinctly different from the drying behaviour of related concrete materials containing aggregates. PMID- 9875608 TI - Analysis of the anisotropic dimension in the RIACT (II) multiple quantum MAS NMR experiment for I = 3/2 nuclei. AB - Numerical simulations of the anisotropic dimension of the Rotation-Induced Adiabatic Coherence Transfer (RIACT) MQ experiment have been performed as a function of the asymmetry parameter, eta, for different spacings between the triple quantum (3Q) excitation and 3Q to single quantum (1Q) reconversion pulses. Large distortions of the spectra are observed, in comparison to the spectra obtained with 1D MAS NMR methods. The method is very sensitive to the relative orientation of the quadrupolar tensor and rotor axis, and signal can only be obtained from a maximum of 60% of the powder. The intensity varies with the spacing between the two pulses, t1, reaching a minimum of 30% when t1 is either a multiple of a full rotor period (n tau(r)) or for (n + 1/2) rotor periods, for pulse lengths (tau1SL) of a quarter of a rotor period. The maximum of 60% is obtained when t1 = n tau(r) - tau1SL. Experimental spectra were acquired for anhydrous Na2HPO4. Good fits were obtained between the experimental and simulated spectra, even for the non-rotor synchronized experiment, by choosing fixed values of t1. The simulations allowed the quadrupole coupling constants and asymmetry parameters to be extracted from the experimental data. PMID- 9875609 TI - Distribution of principal values of the 31P NMR chemical shift tensor in phosphate glasses. AB - The static line shapes of the powder pattern of the 31P nuclear magnetic resonance (NMR) chemical shift (CS) tensor in phosphate glasses can only be explained by using individual distributions parameters for the principal axes. A model is discussed, which assumes a linear dependency of such distributions from the principal values themselves. Furthermore, the influence of such distributions on magic angle spinning (MAS) spectra is investigated. PMID- 9875610 TI - Structural study of a lead-barium-aluminum phosphate glass by MAS-NMR spectroscopy. AB - The 31P- and 27Al-MAS-NMR spectra of a lead-barium-aluminum phosphate glass were measured and analyzed, and the short range structure of the glass are discussed. The 31P-MAS-NMR spectrum of the glass studied has a single isotropic peak at 4.76 ppm, indicating that orthophosphate species are the dominant P-sites in the glass. The 27Al-MAS-NMR spectrum of the glass shows that the aluminum ions present in the glass are in 4-, 5-, and 6- fold coordination with oxygen, and that the Al(OP)4 is the dominant moiety for the glass, suggesting the formation of orthophosphate species in the glass. PMID- 9875611 TI - 1H-13C-27Al triple resonance transfer of populations in double resonance experiments for the detection of 13C-27Al dipolar interactions. AB - A 13C [1H] CPMAS [27Al] TRAPDOR NMR experiment is reported with the aim of detecting 13C-27Al proximities and distances in solids. The 13C and 27Al pulses are applied to the same probe channel, because their resonance frequencies lie extremely close to each other. The study of the heteronuclear dipolar interaction between these two nuclei, which are of fundamental importance in solid state science, is not possible with standard double resonance approaches. Results are presented for the model compound aluminum lactate. The 13C signals for the three lactate carbons show different magnitudes of the TRAPDOR effect, in excellent agreement with their mean Al-C distances, calculated from crystal structure data. The results illustrate the feasibility of this method for the study of systems where the interaction of organic and inorganic fractions is directing the structure (template/zeolite) or controlling the catalytic efficiency (organic reactant/catalytically active sites in zeolites or clays). PMID- 9875612 TI - Familial cavernous angioma: an unknown, known disease. PMID- 9875613 TI - Anticipation in familial cavernous angioma: ascertainment bias or genetic cause. AB - OBJECTIVES: Anticipation has been linked to unstable trinucleotide repeats in many neurological disorders. We examined the hypothesis of genetic anticipation in familial cavernous angioma (FCA) of the central nervous system. MATERIAL AND METHODS: The mean ASO of affected individuals was compared between successive generations in 55 families. Intergenerational pair-wise comparisons were employed to avoid several ascertainment biases. Regarding severity of disease both type of manifestation and number of cavernous angiomas were compared between generations. RESULTS: The mean ASO decreased significantly both from the first to the second generation (31.6 vs 17.8 years; P = 0.000) and from the second to the third generation (17.8 vs 6.7 years; P = 0.002). The pair-wise comparisons also showed significantly earlier ASO. No clear evidence for anticipation with regard to severity of disease was found. CONCLUSIONS: Molecular genetic studies will determine whether trinucleotide repeats are the underlying mechanism for our observation of anticipation in FCA. PMID- 9875614 TI - Focal, continuous spikes suggest cortical developmental abnormalities. Clinical, MRI and neuropathological correlates. AB - OBJECTIVES: Widespread use of MRI now gives us increased insights into the different expressions of malformations of cortical development (MCD). The heterogeneity of these disorders are reflected by their varied clinical and neuroimaging features. Characteristic and intense scalp EEG abnormalities have been described in some patients. MATERIAL AND METHODS: We report the MRI and clinical findings of 3 adult patients (age 32-36) with a peculiar EEG pattern of distinct, localized, fast, continuous spiking. These patients represent all patients with such EEG findings that have been recognized by the first author during 9 years. RESULTS: MRI showed MCDs in all, respectively hemimegalencephaly, a subcortical heterotopion, and a focal cortical dysgenesis. The EEG findings had been stable since childhood and were posteriorly located. Two patients had fairly well controlled epilepsy in adult age. The third patient was incapacitated by persistent seizures and was treated with surgery. Histologically cortical dysplasia with neuronal clusters was found in this patient. Variable degrees of cognitive dysfunction were present in all. CONCLUSION: Focal, continuous, fast spiking is an unusual scalp EEG pattern. It is not an inevitable sign of severe epilepsy. It may suggest an MCD. It is not yet clear to what extent such findings are predictive of a dysgenetic etiology of epilepsy. PMID- 9875615 TI - Quantitative EEG in acute mountain sickness. AB - OBJECTIVES: To investigate if the EEG response at moderate altitude may predict a person's tolerance to acute mountain sickness (AMS). MATERIALS AND METHODS: Frequency analysis (QEEG) of tape-recorded ambulatory EEG was performed in 6 climbers during a mountaineering expedition to 7546 m above sea level. The QEEG response in climbers, measured at sea level, at 4500 m, and at 1800 m 1-4 days after maximal altitude exposure, was compared to the change observed during consecutive sea level recordings in 10 control subjects. RESULTS: Three climbers experienced slight (grade 1) AMS symptoms both at 4500 m and at maximal altitude exposure (Group 1). Three other climbers (Group 2) had no symptoms at 4500 m, but they developed AMS (grades 1, 2, or 3) at maximal altitude. Alpha amplitudes were higher at 4500 m in group 1 climbers, while it was lower in group 2 climbers compared to the sea level recording. Significant time x group interactions in ANOVA were found for delta (P = 0.005), theta (P = 0.001) and alpha (P = 0.001) amplitude, indicating that QEEG amplitudes decreased significantly at high altitude in group 2 climbers. CONCLUSION: The QEEG response to moderate hypobaric hypoxia is not uniform, but the direction of QEEG amplitude change, particularly in the alpha band, may possibly predict the risk of developing AMS. PMID- 9875616 TI - An epidemiological study of multiple sclerosis in central Sardinia, Italy. AB - OBJECTIVES: To verify morbidity estimates in central Sardinia, Italy. METHODS: A prevalence study was performed in the province of Nuoro, Central Sardinia, which has a population of 273,768 inhabitants (135,383 men and 138,385 women). A complete enumeration approach was adopted by using all possible case-collection sources. RESULTS: On prevalence day, December 31, 1993, 394 subjects (124 men and 270 women) living in the study area were known to suffer from definite and probable MS, giving a crude prevalence rate of 143.9 cases per 100,000 people, 91.6 for males and 195.11 for females. The crude prevalence estimated on December 31, 1985, based on 282 MS cases alive in the study area, was 102.94 per 100,000. CONCLUSION: This study reinforced central Sardinia's position as a high and rising prevalence area for MS. PMID- 9875617 TI - A single subtype of Epstein-Barr virus in members of multiple sclerosis clusters. AB - OBJECTIVES: Epidemiological studies strongly indicate an infectious involvement in multiple sclerosis (MS). Epstein-Barr virus (EBV), to which all multiple sclerosis patients are seropositive, is also interesting from an epidemiological point of view. We have reported a cluster of MS patients with 8 members from a small Danish community called Fjelso. To further evaluate the role of EBV in MS we have investigated the distribution of EBV subtypes in cluster members and in control cohorts. MATERIALS AND METHODS: Blood mononuclear cells were isolated from cluster members, unrelated MS patients, healthy controls, including healthy schoolmates to the Fjelso cluster patients and finally from persons with autoimmune diseases in order to investigate the number of 39 bp repeats in the EBNA 6-coding region in the EBV seropositive individuals. RESULTS: We observed a preponderance of the subtype with 3 39 bp repeats in the EBNA 6-coding region both in the MS patients and the healthy controls. In the Fjels cluster all 8 cluster members were harbouring this subtype, which is significantly different from the finding in healthy controls (n = 16), which include 8 schoolmates to the cluster members and 8 randomly selected healthy persons (Fischer's exact test P = 0.0047), and also compared to all non-clustered individuals studied (P = 0.017). CONCLUSION: Infection with the same subtype of EBV links together the 8 persons from the Fjelso cluster who later developed MS. This finding adds to the possibility that development of MS is linked to infection with EBV. PMID- 9875618 TI - Sexual dysfunction in stroke patients. AB - OBJECTIVES: A marked decline in sexual activity has been reported in stroke patients. Little information, however, is available on clinical correlates of sexual dysfunction, changes in the sexual life of spouses of stroke patients, and abnormalities of nocturnal penile erections. MATERIAL AND METHODS: In this prospective 6-month follow-up study, we assessed the impact of stroke on libido, sexual arousal, coital frequency and satisfaction with sexual life in 50 stroke patients (38 men, 12 women, aged 32 to 65 years) and in their spouses. We also measured nocturnal penile erections of the male patients using a strain gauge attached around the penis. RESULTS: All the analyzed aspects of sexuality, i.e., libido, coital frequency, erection, ejaculation, vaginal lubrication, orgasm and satisfaction with sexual life, were commonly decreased as a consequence of stroke both in the patients and in their spouses. Fourteen (28%) patients at 2 months post-stroke and 6 (14%) patients at 6 months had ceased having sexual intercourse. Nocturnal erections at 2 months post-stroke were normal in 17 (45%) of the 38 male patients and impaired in 21 (55%) patients, but all of the patients had some nocturnal erections. Sexual dysfunction correlated significantly (P<0.05) with the presence of the sensory hemisyndrome. CONCLUSIONS: Sexual dysfunction and dissatisfaction with sexual life seems to be common both in male and female stroke patients and in their spouses. Decreased libido, sexual arousal and satisfaction are related particularly to the presence of the hemisensory syndrome. The etiology for sexual dysfunction after stroke is multifactorial including both organic and psychosocial factors. PMID- 9875619 TI - Blood-brain barrier damage in reperfusion following ischemia in the hippocampus of the Mongolian gerbil brain. AB - Vascular permeability to intravenously injected horseradish peroxidase (HRP) was qualitatively examined in the hippocampus of ischemic Mongolian gerbil brains by light and electron microscopy. After 30 min of right common carotid artery occlusion followed by 90 min of reperfusion, the animal was perfused with a fixative and killed. Before the perfusion of the fixative, HRP was injected into the femoral vein. HRP was visualized with tetramethyl benzidine (TMB) and diamino benzidine (DAB) for light and electron microscopy, respectively. Staining reaction with TMB for HRP appeared in medial or dorsal portions of the operated side of the hippocampus, especially around some vessels along the hippocampal fissure. Ultrastructural examination in the vessels along hippocampal fissure revealed that the endothelial cytoplasm contained HRP-filled vesicles or vacuoles in close proximity to the basal lamina, and seemed to be slightly electron-dense. Swollen pericytes, swollen astrocytic foot processes and perivascular cells with HRP-filled cytoplasm were also observed in that area. In this study, it was clearly demonstrated that intravascular macromolecules leaked transendothelially, through vessel walls in the hippocampal fissure, from the blood stream in the medial portions of the hippocampus during reperfusion following ischemia. These findings suggest that the blood-brain barrier in some vessels along the hippocampal fissure in the medial parts of the hippocampus is more vulnerable to ischemic insults than those in other brain areas. PMID- 9875620 TI - Conditioning effect on the long latency potentials in the lower limb to transcranial magnetic stimulation. AB - OBJECTIVES: We used an electrical conditioning stimulation followed by transcranial magnetic stimulation (TMS) to facilitate the occurrence of long latency potentials (LLPs) in order to study the relationship between primary motor evoked potentials (MEPs) and LLPs in the lower limbs. MATERIALS AND METHODS: The study group included 6 healthy subjects, 1 patient with right thalamic infarction, and 3 patients with spinal cord injuries. The subjects were subjected to electrical conditioning (C) stimulation delivered to the left big toe at 250 Hz in a train of pulses of 20 ms duration prior to TMS (T) from 0 to 150 ms at an increment of 10 ms. The surface electromyographic signals were recorded at the tibialis anterior and gastrocnemius medialis for 400 ms. RESULTS: The C-T test facilitated both primary MEPs and LLPs with a pattern similar to the primary MEPs of its antagonist. There was no facilitation of the primary MEPs or LLPs in the affected limb of patients with thalamic or spinal cord lesions. CONCLUSION: At appropriate C-T interval, LLPs could be consistently provoked by TMS. The LLPs were absent in the patients with thalamic infarction and spinal cord injuries. It suggests that LLPs might be provoked through a supraspinal control. PMID- 9875621 TI - The B-cell repertoire in myasthenia gravis includes all four acetylcholine receptor subunits. AB - By enumerating cells secreting IgG antibodies of particular specificities using an enzyme-linked immunospot (ELISPOT) assay, the B-cell responses to Torpedo acetylcholine receptor (AChR) and its alpha-, beta-, gamma- and delta-subunits in peripheral blood from patients with myasthenia gravis (MG), and controls with other neurological diseases (OND) as well as healthy subjects were determined. Compared to controls, the patients with MG had elevated numbers of B cells secreting antibodies against AChR and its alpha-, beta-, gamma- and delta subunits in peripheral blood in parallel. The mean numbers of anti-AChR antibody secreting cells were about 17 per 10(5) blood MNC, and for the subunits 10 to 15 in MG patients, compared to between 0.8 and 1.9 per 10(5) blood MNC in OND patients, and 0.1 to 0.3 in healthy controls. Such B cells detected in controls probably represent naturally occurring B cells responded to AChR and its subunits. The finding that most (60%) MG patients had B cells predominantly recognizing the alpha-subunit may be an indirect argument for the existence of a main immunogenic region (MIR). In the remaining 40% of patients with MG the predominant B-cell responses were directed to beta-, gamma- or delta-subunit. The data suggest that all four AChR subunits may function as strong immunogens in MG, though the alpha-subunit may be the major immune target in a substantial proportion of MG patients. PMID- 9875622 TI - Clinical and genetic studies of spinocerebellar ataxia type 2 in Japanese kindreds. AB - OBJECTIVES: We report the results of clinical and genetic studies from 2 related Japanese kindreds with spinocerebellar ataxia type 2 (SCA2). MATERIAL AND METHODS: Family A showed 19 patients through 4 generations, while family B showed 6 patients, including dizygotic twin brothers, through 3 generations. We performed clinical, radiological, neurophysiological, and genetic analyses in the family members. RESULTS: Neurologic analysis of 13 affected patients revealed a mean age at onset of 43.5 years. The most common neurologic finding was cerebellar ataxia with deep sensory disturbance. Slow saccades was found only in the younger patients below age 35 years. Nerve conduction studies revealed subclinical sensory neuropathy. Brain MRI showed the presence of pontocerebellar atrophy. Genetic study using PCR revealed that all affected patients had an expanded CAG allele in the ataxin-2 gene, which led to a final diagnosis of SCA2. CONCLUSION: SCA2 may be more clinically heterogeneous than previously thought. PCR is useful in differentiating SCA2 from other types of inherited ataxia. PMID- 9875623 TI - The effect of glucocorticoids in the normal cerebral hemisphere of brain tumour patients. AB - OBJECTIVES: To study the potential effects of glucocorticoids (GC) in normal white matter (WM) of the contralateral cerebral hemisphere in brain tumour patients in a prospective protocol. MATERIALS AND METHODS: Using MR relaxation time (RT) imaging (T1 and T2-maps), 190 different scanning sessions were performed on 42 brain tumour patients before and after initiation of GC treatment. RT imaging correlates closely with in vivo tissue water content (reported estimation accuracy error <4%). Repetitive studies were performed in order to study changes as a function of time. RTs were measured in the contralateral frontal and occipital white matter (WM). RESULTS: The mean change in T1 after initiation of GC treatment was 1.6+/-1.7% (P = 0.24), and 1.8+/-3.8% in T2. The mean coefficient of variation for the pooled data in all patients was 1.4% for both T1 and T2. There were no differences in effect between different histological types of tumours or sex. CONCLUSION: GC treatment does not influence in vivo cerebral water content expressed as relaxation times of apparently normal WM. The neurological effects in brain tumour patients are normally evident within 24 h, but do not seem to correlate with changes in brain water content. The normal biological variation in RTs (and thereby in vivo tissue water content) in normal WM of brain tumour patients is very small and approaches the expected fluctuations in the MRI measurement technique. PMID- 9875624 TI - Akinetic mutism: a report of three cases. AB - OBJECTIVES: To report 3 new cases of akinetic mutism, a clinical syndrome defined by silent immobility with preserved visual alertness not accountable by lesion of the areas and/or effector pathways of speech and voluntary movements. MATERIAL AND METHODS: Anatomopathological studies were performed in Cases 1 and 2; clinical follow-up, EEG, angiography and CT scans in Case 3. RESULTS: Case 1: Bipallidal necrosis; Case 2: Left pallidal necrosis with right frontoparietal cortico-subcortical infarction; Case 3: Striato-capsular infarction on the left side, involving the caudate nucleus and the anterior arm of the internal capsule, together with obstructive hydrocephalus. CONCLUSION: The roles of both globus pallidus and prefrontostriatal circuits in the onset of voluntary movements are discussed. PMID- 9875625 TI - Post-dural puncture related complications after diagnostic lumbar puncture, myelography and spinal anaesthesia. AB - OBJECTIVES: This study was conducted to investigate complications after dural puncture. MATERIAL AND METHODS: A 15 months' prospective observation study of routine clinical practice with dural puncture at a university hospital was conducted. Quincke spinal needles 0.90 to 1.0 mm O.D. (20-19 g) were used for diagnostic lumbar puncture, 0.70 mm O.D. (22 g) for myelography and 0.40 to 0.50 mm O.D. (27-25 g) for spinal anaesthesia. A questionnaire about post-puncture discomfort was given to the patients, to be returned after 1 week. RESULTS: Of 679 questionnaires 537 (79.1%) were returned. Discomfort was experienced by 53.8% of the patients, most often after diagnostic lumbar puncture and myelography. The difference in incidence of headache after diagnostic lumbar puncture and myelography compared with spinal anaesthesia were 27.9% (95% CI: 18.6 to 37.2) and 18.3% (95% CI: 9.1 to 27.5). CONCLUSION: Small diameter and atraumatic spinal needles will reduce patients' discomfort after dural puncture. PMID- 9875626 TI - A clinicopathological observation of Nyssen-van Bogaert syndrome with second motor neuron degeneration: two distinct clinical entities. AB - OBJECTIVES: To report a novel clinicopathological observation of Nyssen-van Bogaert syndrome. To compare this observation with those previously reported. To discuss the nosological entity of this syndrome. To define the exact etiopathogenic mechanism of the neurogenic amyotrophy occurring at the late stage of the disease. MATERIALS AND METHODS: The patient was a 16-year-old girl who developed loss of vision and deafness at the age of 8. Ataxia with slight cerebellar signs were present by the age of 14. Over the next 2 years, she developed distal weakness and wasting of the legs with depressed ankle reflexes. She died at the age of 16. Deparaffinized sections of the brain, the brain stem, the cerebellum and the spinal cord were stained with haematoxylin & eosin (H&E), Nissl, Woelcke, Bodian, periodic acid-schiff (PAS), Sudan Black and Kluver Barera. Antibodies anti-GFAP, anti-MPB and anti-neurofilaments were used for immunohistochemical stainings following the avidin-biotin-peroxydase complex (ABC) methods. RESULTS: The clinical pictures in our patient are similar to those previously reported in juvenile patients with optico-cochleo-dentate syndrome. Pathological study of the nervous system confirmed the diagnosis of Nyssen-van Bogaert syndrome and also showed a severe anterior horn, posterior horn and Clarke's column nerve cell degeneration with anterior root atrophy. CONCLUSION: From these clinical and pathological data, the authors suggest to include Nyssen van Bogaert syndrome among the group of multiple system atrophy, propose to divide this syndrome into 2 forms (an early infantile form and a juvenile form) and consider that the neurogenic amyotrophy occurring at the late stage of the disease in juvenile and adult patients is mainly caused by the second motor neuron involvement. PMID- 9875628 TI - Chronic inorganic mercury induced peripheral neuropathy. AB - We report the clinical features, electrophysiological studies, and morphometric analysis of sural nerve pathology in a patient with polyneuropathy due to inorganic mercury intoxication. He developed slowly progressive generalized paralysis of all limbs after 3 months ingestion of herb drugs which contained mercuric sulfate. Electrophysiologic studies revealed axonal polyneuropathy involving both motor and sensory fibers. Sural nerve biopsy demonstrated axonal degeneration with demyelination and a predominant loss of large myelinated fibers. His muscle strength showed only mild improvement after 2 years' follow up. We concluded that inorganic mercury exposure may induce severe axonal sensorimotor polyneuropathy in humans and that neurological deficits may persist in severe cases. PMID- 9875627 TI - A case of myokymia-cramp syndrome successfully treated with gabapentin. AB - We report a case of "myokymia-cramp syndrome", a rare and benign disease consisting of muscular cramps in the upper and lower limbs associated with generalized myokymias. In our patient, cramps had been present since adolescence and had increased in frequency and duration during the last 3 years, occurring about 8 or 9 times a day. Cramps were mainly nocturnal and also precipitated by walking; a previous treatment with carbamazepine did not improve the symptoms. Gabapentin therapy proved to be very satisfactory in relieving muscular cramps with a relatively low dosage (600 mg/day) and without any remarkable side effects. The possible interpretation of the mechanism involved in gabapentin induced relief of cramps is discussed. Gabapentin should be considered as a safe alternative treatment for muscular cramps. PMID- 9875629 TI - Standardized risk comparisons of neurological disorders in different populations. PMID- 9875630 TI - Progressive supranuclear palsy revisited. AB - Current understanding on the historic, epidemiologic, genetic, clinical, neuropathologic, neurochemical, diagnostic and therapeutic aspects of progressive supranuclear palsy (PSP) are revisited. In addition, new research directions are described. PMID- 9875631 TI - The distribution of HBV, HCV and HGV among livers with fulminant hepatic failure of different aetiology. AB - BACKGROUND/AIMS: The aim of the study was to assess the impact factor of HCV and HGV in fulminant hepatic failure. METHODS: The 5'-untranslated regions of HCV RNA and HGV RNA and a segment of the core antigen sequence of HBV were amplified after extracting the nucleic acids from snap-frozen tissue aliquots from explanted livers of 26 consecutive patients undergoing orthotopic liver transplantation for fulminant hepatic failure preoperatively diagnosed as either autoimmune (n=2), HAV/HBV (n=8), toxic (n=4) or aetiologically unknown (n=12). RESULTS: HCV RNA was detected in five of 26 (19.2%) livers with fulminant hepatic failure. All five HCV RNA-positive livers belonged to the group of non-toxic, non autoimmune liver failure (n=20), three of them were found in the group of liver failure with unknown aetiology (n=12) and two in the group of HBV-associated liver failure (n=7), making an HCV incidence of 25%, 25% and 28.6%, in the different groups, respectively. HGV RNA was detected in 10 of 17 (58.8%) explants and in all four groups of fulminant hepatic failure as defined preoperatively. HBV DNA was identified in six livers of 26 patients (23.1%) with fulminant hepatic failure. Neither HCV RNA nor HBV DNA was detected in the livers of patients with toxic or autoimmune fulminant hepatic failure. CONCLUSIONS: These results indicate that HBV and HCV, but not HGV, play an aetiologic role in fulminant hepatic failure. HCV-positive cases were concentrated either in the group of otherwise unexplained fulminant hepatic failure or in the group of HBV fulminant hepatic failure. HGV-positive cases, on the other hand, were found within all four preoperatively defined groups, indicating a role as cofactor rather than as single aetiologic agent. PMID- 9875632 TI - Transactivation of the human MDR1 gene by hepatitis B virus X gene product. AB - BACKGROUND/AIMS: Persistent hepatitis B virus (HBV) infection may cause hepatocellular carcinoma. Patients with hepatocellular carcinoma are characterized by nonresponsiveness to chemotherapeutic agents. While many studies have been devoted to understanding the hepatocarcinogenesis mechanism of HBV, the possible relationship between HBV and the drug sensitivity phenotype of cancer cells has rarely been addressed. The hepatitis B virus X gene encodes a transcription transactivator which has been suggested to be a potential factor in viral hepatocarcinogenesis. The role of HBV pX in mediating the drug resistance phenotype of hepatoma cell lines was examined in this study. METHODS: Standard transfection and chloramphenicol acetyltransferase assay were utilized to examine the effect of HBV pX transactivator on a reporter gene under the control of the human multidrug resistance (MDR) 1 upstream regulatory elements. Selected Hep G2 clones with or without HBV pX expression were tested for their sensitivity towards various anti-cancer agents by utilization of MTT assay. RESULTS: The expression of HBV pX in both Hep G2 (p53+) and Hep 3B (p53-) cells resulted in transactivation of the reporter gene under control of the human MDR1 upstream regulatory elements. Northern blot analysis indicated that expression of the endogenous MDR1 gene was also elevated in Hep G2 clones with HBV pX expression. Decreased drug sensitivity towards adriamycin, vinblastine, and VP-16 was observed in Hep G2 clones with HBV pX expression. CONCLUSIONS: HBV pX can transactivate the MDR1 gene. Drug sensitivity was altered in Hep G2 cells with HBV pX expression. PMID- 9875633 TI - Clonality and specificity of cryoglobulins associated with HCV: pathophysiological implications. AB - BACKGROUND/AIMS: Hepatitis C virus (HCV) infection plays a central role in the pathogenesis of mixed cryoglobulinemia through molecular mechanisms which remain to be elucidated. The aim of this study was to investigate the role of antibody responses to HCV in the pathogenesis of cryoglobulinemia through characterization of the anti-HCV specificity and immunochemical characteristics of the immunoglobulins involved in cryoprecipitation. METHODS: Sera from 50 consecutive patients with chronic HCV infection (RNA positive) were screened for the presence of cryoglobulins. The two major components of cryoprecipitates, IgM rheumatoid factors and IgG, were separated by high performance liquid chromatography and analyzed for immunochemical composition by immunoblotting and antibody specificity by ELISA and immunoblotting using recombinant HCV proteins and synthetic peptides as antigens. RESULTS: Cryoprecipitates were observed in 27 patients and characterized by immunofixation: 13 (48%) were classified as type II and 14 (52%) as type III. Monoclonal immunoglobulins were detected by immunoblotting in 20 cryoprecipitates: IgM in 14 samples and IgG in 14, with a clear preponderance of IgG3 (12/14). Specificity studies on sera and purified IgM and IgG fractions from cryoprecipitates revealed enrichment in cryoglobulins, predominantly polyclonal IgG1, reactive with the HCV structural proteins, whereas specificities for nonstructural viral proteins were relatively less represented compared to whole serum. No restricted pattern of fine specificity was observed. IgG3 subclass was apparently not involved in HCV nucleoprotein binding. CONCLUSIONS: Our findings do not support a direct link between monoclonal cryoglobulins and immune response to HCV According to the proposed pathogenetic model, HCV infection can induce the formation of cryoprecipitable rheumatoid factors, sustain their production, and eventually lead to monoclonal B-cell expansion through several cooperative mechanisms. PMID- 9875634 TI - Comparison of serum and liver hepatitis C virus quasispecies in HCV-related hepatocellular carcinoma. AB - BACKGROUND/AIMS: The hepatitis C virus (HCV) genome consists of quasispecies populations of heterogeneous variants, especially in the hypervariable region. To assess the profiles of viral quasispecies in HCV-related hepatocellular carcinoma, we studied the viral population patterns in serum and liver tissues of 13 HCV-positive patients with hepatocellular carcinoma developed on cirrhotic and non-cirrhotic livers (5 and 8 cases, respectively). METHODS: HCV genome heterogeneity was analyzed by polymerase chain reaction-mediated single-strand conformation polymorphism analysis, which showed multiple DNA bands representing different hypervariable region sequences. RESULTS: The HCV populations were different between tumorous and nontumorous tissues in 3/5 hepatocellular carcinomas with cirrhosis and in 6/8 without cirrhosis. At least one or more than one common band was detected in both compartments in all but one case. No significant differences in the complexity of HCV quasispecies were found in hepatocellular carcinoma with or without underlying cirrhosis. Comparison of the HCV quasispecies profiles in serum and liver tissues showed a different distribution of HCV variants between these two compartments in 6/7 patients. In four cases, both common and compartmentalized sequences were detected, whereas in two cases, both without cirrhosis, the HCV population in serum was completely different from that found in the liver. CONCLUSIONS: These results suggest that the complexity of HCV populations is influenced by the presence of hepatocellular carcinoma rather than by the severity of the underlying chronic liver disease. The different quasispecies patterns found in serum and liver may reflect different biological properties of circulating and intrahepatic HCV particles or the existence of extrahepatic sites of replication. PMID- 9875635 TI - Impact of hepatitis G virus co-infection on the course of hepatitis C virus infection before and after liver transplantation. AB - BACKGROUND/AIMS: Hepatitis G virus (HGV), a new RNA virus that is parenterally transmitted, has frequently been found in patients with chronic hepatitis C (HCV) infection but its role in chronic liver disease is unknown. The purpose of this study was to determine the prevalence of HGV infection in transplantation patients infected with hepatitis C and to assess the impact of HGV co-infection on the course of HCV infection after liver transplantation. METHODS: Eighty-nine liver transplantation recipients with persistent hepatitis C viremia detected by polymerase chain reaction (PCR) were evaluated. Serum samples were tested before and after liver transplantation for HGV RNA by two different PCR methods: LC assay (Abbott Laboratories) and an RT-PCR procedure which we developed using the silica gel technique for extraction of the HGV RNA. E2 antibodies were detected before orthotopic liver transplantation by an EIA-test. HCV RNA was quantified by branched DNA assay, and HCV genotype was determined. A mean of nine liver biopsy specimens were examined for each patient and the severity of the lesions was compared in HCV-positive patients with or without HGV co-infection. RESULTS: The concordance between the two HGV RNA detection methods was excellent and the reproducibility of our RT-PCR procedure was confirmed. The prevalence of pretransplantation and posttransplantation HGV infection was 11% and 19%, respectively. Pretransplantation HGV infection was positively correlated with posttransplantation HGV infection (p<0.001). Before transplantation the E2 antibodies seroprevalence was 34%. Seven patients became HGV RNA positive after transplantation, but all of them were negative for E2 antibodies. Among the patients who remained RNA negative after liver transplantation, 40% were positive for E2 antibodies (p=0.04). Pretransplantation clinical features (except AST mean value) were not different in patients with HCV and HGV co-infection and those with HCV only. After a mean follow-up of 34 months (range: 6 to 70), 67/89 (75%) patients developed histological features of recurrent hepatitis but the frequency of the occurrence of graft hepatitis was not different between HGV/HCV co infected patients and those with HCV alone (p=0.89). The mean interval from orthotopic liver transplantation to recurrence was 12.2 months (range: 3-63), which was not different for HVG/HVC-co-infected patients and HCV-infected patients. The histological severity of posttransplantation liver disease, and the graft and patient survival were not different for patients with and without HGV co-infection. CONCLUSIONS: Our results suggest the general persistence of HGV infection after liver transplantation, but HGV co-infection did not appear to influence the posttransplantation course of HCV infection. Before transplantation the prevalence of E2 antibodies was 34%, and our data clearly indicate that E2 antibodies were protective against HGV infection. PMID- 9875636 TI - Expression and regulation of intercellular adhesion molecule-1 (ICAM-1) in organotypic cultures of rat liver tissue. AB - BACKGROUND/AIMS: The objective of the present study was to analyze the expression and regulation of intercellular adhesion molecule-1 (ICAM-1) in organotypic cultures of rat liver slices, which preserve the normal microenvironment of liver cells. METHODS: Rat liver slices were maintained in culture for 15 min to 24 h and examined for ICAM-1 expression by immunohistochemistry and Western blotting in basal conditions and after stimulation with 1000 IU/ml interferon-gamma (IFNgamma), 1000 IU/ml tumor necrosis factor-alpha (TNF alpha) and 50 microg/ml endotoxin. Immunohistochemical results were evaluated using a semiquantitative scoring system. RESULTS: In uncultured slices, ICAM-1 was not detected on hepatocytes. In unstimulated liver slices maintained in organotypic culture, ICAM 1 was induced at the surface of scattered hepatocytes (score at 15 min, 0.33+/ 0.47 and at 24 h, 1.17+/-0.69). After 4 h of stimulation, a significant increase in ICAM-1 expression by hepatocytes and adjacent sinusoidal cells, but not by intra-hepatic biliary epithelial cells, was observed for IFNgamma (score: 2.35+/ 0.47) and endotoxin (score: 2.67+/-0.47), but not with TNF alpha (score: 0.66+/ 0.47). After 24 h of stimulation, a further increase in the extent of ICAM-1 expression by hepatocytes was observed for IFNgamma (score: 3.67+/-0.47) and endotoxin (score: 4.0+/-0.0), and a significant overexpression of ICAM-1 by hepatocytes was detectable after treatment with TNF alpha (score: 3.67+/-0.47). CONCLUSIONS: In rat liver organotypic cultures, TNF alpha, IFNgamma and endotoxin induce the expression of ICAM-1 in hepatocytes and adjacent sinusoidal endothelial cells, but not in portal tracts. PMID- 9875637 TI - HVJ-liposome mediated gene transfer into hepatocytes in vivo. AB - BACKGROUND/AIMS: The efficient transduction of appropriate target cells will be critical for gene therapy. We evaluated the suitability of hemagglutinating virus of Japan (HVJ)-liposome-mediated gene transfer for gene therapy of liver diseases. METHODS: The Escherichia coli beta-galactosidase (beta-gal) gene was introduced into rat liver by HVJ-liposome to examine gene transfer efficacy and persistence of expression with or without partial hepatectomy prior to transfection. RESULTS: About 30% of hepatocytes were transduced after portal vein injection. Gene expression was transient, with only 2% of hepatocytes expressing beta-gal after 4 weeks. However, partial hepatectomy performed 24 h prior to injection resulted in persistently high levels of beta-gal for 4 weeks after injection. A 247-bp beta-gal polymerase chain reaction fragment transcript was detected in livers of transfected rats, but not in livers of control rats. The rat livers following gene transfer were histologically normal, and serum glutamic pyruvic transaminase was not found to be elevated in rats. CONCLUSIONS: Our results demonstrate that HVJ-liposome-mediated gene transfer produced high gene transduction and persistent gene expression in the liver. PMID- 9875638 TI - Expression of different isoforms of TGF-beta and the latent TGF-beta binding protein (LTBP) by rat Kupffer cells. AB - BACKGROUND/AIMS: Kupffer cells (liver resident macrophages) make an important contribution to the perpetuation of liver diseases by synthesis and secretion of TGF-beta. In some cell types TGF-beta, is expressed as a large latent complex containing the latent TGF-beta binding protein (LTBP) in addition to the N terminal TGF-beta precursor (latency associated peptide). This study aimed to identify LTBP expression in rat Kupffer cells. METHODS: Cells were isolated from rat liver by collagenase-pronase reperfusion, purified and cultured under standard conditions. TGF-beta and LTBP expression were characterized using alkaline phosphatase-anti-alkaline phosphatase immunostainings, reverse transcription-polymerase chain reaction and immunoprecipitation of metabolically labeled proteins. RESULTS: Immunostainings of Kupffer cells with anti-sera against LTBP-1 (ab 39) and LTBP-2 indicated the expression of both LTBP isoforms in addition to the expression of latency associated peptide and TGF-beta. Transcripts of three LTBP isoforms (LTBP-1,-2,-3) and TGF-beta isoforms (TGF-beta 1,-2,-3) were detectable by reverse transcription-polymerase chain reaction. The LTBP-1D splice variant missing a part of the proteinase sensitive hinge region which has recently been described in hepatic stellate cells is expressed in Kupffer cells, too. Metabolic labeling of Kupffer cells with [35S]-Met/Cys followed by immunoprecipitation of the conditioned media using antisera against LTBP-1 and LTBP-2 indicated the secretion of high molecular mass TGF-beta complexes containing LTBP proteins of 230 and 170 kDa (LTBP-1) or 230 kDa (LTBP 2). CONCLUSION: The results show that Kupffer cells partly synthesize and release TGF-beta as large latent complexes. This requires the extracellular activation of TGF-beta as a prerequisite for receptor binding and cellular signaling. PMID- 9875639 TI - Characterization of cationic amino acid transporter and its gene expression in rat hepatic stellate cells in relation to nitric oxide production. AB - BACKGROUND/AIMS: Nitric oxide is a potent mediator of hepatic sinusoidal hemodynamics and affects hepatic stellate cells (Ito cells, fat-storing cells). Although nitric oxide production may depend on the induction of inducible nitric oxide synthase and on transport of extracellular L-arginine, the precise mechanisms controlling nitric oxide production in stellate cells have not been well characterized. METHODS: Using stellate cells prepared from the male Wistar rat, kinetic analysis of L-arginine transport and reverse transcription polymerase chain reaction for cationic amino acid transporter were carried out. The effect of tumor necrosis factor-alpha and interferon-gamma on L-arginine transport, mRNA expression of cationic amino acid transporter and inducible nitric oxide synthase, and nitric oxide production of stellate cells was assessed. RESULTS: The L-arginine transport system functioning in the transformed hepatic stellate cells was system y+, possibly mediated by cationic amino acid transporter-1 and cationic amino acid transporter-2B (Km approximately 50 microM). Tumor necrosis factor-alpha enhanced cationic amino acid transporter-2B mRNA expression and L-arginine transport, whereas cationic amino acid transporter 1 mRNA expression remained unchanged. Interferon-gamma induced the expression of inducible nitric oxide synthase mRNA without obvious changes in L-arginine transport. Interferon-gamma in combination with tumor necrosis factor-alpha induced nitric oxide production with an enhancement in cationic amino acid transporter-2B mRNA expression, inducible nitric oxide synthase mRNA expression, and L-arginine transport, while extracellular L-lysine competitively inhibited this nitric oxide production. CONCLUSIONS: In transformed hepatic stellate cells, tumor necrosis factor-alpha and interferon-gamma have a crucial role in nitric oxide production, and extracellular L-arginine transport and inducible nitric oxide synthase expression are regulated in a differential cytokine-specific manner. As the estimated Km of L-arginine transporter in transformed hepatic stellate cells is very similar to the physiological L-arginine concentration in portal vein, we assume that increased portal L-arginine concentration may easily affect sinusoidal blood flow through enhancement of autocrine nitric oxide production in transformed hepatic stellate cells of diseased liver. PMID- 9875640 TI - Retinyl palmitate reduces hepatic fibrosis in rats induced by dimethylnitrosamine or pig serum. AB - BACKGROUND/AIMS: Lipid peroxidation has been found to be associated with Ito cell activation. Ito cells are the principal collagen-producing cells and the main storage sites of retinoids. However, the relationship between retinoids and hepatic fibrosis is complex. The aim of this study was to elucidate the role of retinoids as a fibrosuppressant: the effects of retinoids on hepatic fibrosis induced in rats by dimethylnitrosamine or pig serum, as well as on rat Ito cells in primary culture, were examined in order to assess the antioxidant activity of retinoids. METHODS: Male Wistar rats were given a single injection of 40 mg/kg dimethylnitrosamine or 0.5 ml PS twice weekly for 10 weeks. In each model, rats were treated with retinyl palmitate for 2 weeks before hepatotoxin treatments or for the last 2 weeks of the treatments. The cumulative amount of retinyl palmitate administered in each experiment was 2, 10, or 20x10(4) IU/rat. RESULTS: Retinyl palmitate treatment before or after administration of dimethylnitrosamine or pig serum suppressed the induction of hepatic fibrosis, restored hepatic retinyl palmitate levels, prevented increases in hepatic levels of collagen and malondialdehyde, a product of lipid peroxidation, and prevented increases in deposition of type III collagen and the number of alpha-smooth muscle actin (alpha-SMA) positive-Ito cells in the liver. Retinyl palmitate supplementation resulted in a dose-dependent reduction of alpha-SMA expression and an oxidative burst in cultured Ito cells. In addition, retinyl palmitate inhibited Fe2+/adenosine 5'-diphosphate-induced lipid peroxidation in rat liver mitochondria and showed radical scavenging activity. CONCLUSIONS: These findings suggest that retinyl palmitate may suppress the induction of hepatic fibrosis, at least in part, by the inhibition of Ito cell activation through its antioxidant activity. PMID- 9875641 TI - Characteristics of hepatocellular carcinoma in Italy. AB - BACKGROUND/AIMS: This study aimed to assess the main features of hepatocellular carcinoma at the time of diagnosis in Italy, particularly in relation to the presence or absence of underlying cirrhosis, hepatitis virus marker patterns, age of the subjects and alpha-foetoprotein values. METHODS: A total of 1148 patients with hepatocellular carcinoma seen at 14 Italian hospitals in the 1-year period from May 1996 to May 1997 were the subjects of this prevalence study. Both newly diagnosed cases (incident cases) and cases diagnosed before May 1996 but still attending the hospitals during the study period (prevalent cases) were included. RESULTS: We found that 71.1% of cases were positive for hepatitis C virus antibodies but negative for HBsAg; in contrast, 11.5% were negative for anti-HCV but positive for HBsAg; 5.3% were positive for both markers; and 12.1% were negative for both viruses. The mean age of detection was over 60 years, with a younger mean age in HBsAg-positive compared to anti-HCV-positive patients (59.3 years vs. 65.6 years, p<0.01). The male-to-female ratio among HBsAg-positive patients was 10.4:1, in contrast to 2.8:1 among anti-HCV-positive patients (p<0.01). The majority of cases (93.1%) had underlying cirrhosis. Cirrhotic patients were more likely to be anti-HCV positive than non-cirrhotic cases (73.2% vs 43.9%; p<0.01); conversely, absence of hepatitis virus markers was more frequently observed in the non-cirrhotic than in the cirrhotic population (40.9% vs. 10.0%; p<0.01). Overall, the alpha-foetoprotein level was altered (>20 ng/ml) in 57.9% of patients; only 18% of cases presented diagnostic (>400 ng/ml) values. Anti-HCV positivity (O.R. 2.0; CI 95%=1.3-3.1) but not HBsAg positivity (O.R. 1.0; CI 95%=0.6-1.8) was shown to be an independent predictor of the likelihood of altered alpha-foetoprotein values by multivariate analysis. CONCLUSIONS: These findings point to differences in the characteristics of the populations infected by hepatitis B and hepatitis C. Factors other than the hepatitis viruses are important in non-cirrhotic patients. A change in the relative prevalence of hepatitis virus markers among hepatocellular carcinoma cases was demonstrated, reflecting a significant change in the rate of HBV endemicity in the Italian population. Finally, the increased trend in the mortality rate from liver cancer in Italy from 4.8 per 100,000 in 1969 to 10.9 in 1994 may reflect the large cohort of subjects infected with HCV via the iatrogenic route during 1950s and 1960s when glass syringes were commonly used for medical treatment. PMID- 9875642 TI - Arterial chemoembolization before liver transplantation in patients with hepatocellular carcinoma: marked tumor necrosis, but no survival benefit? AB - BACKGROUND/AIMS: Hepatic artery chemoembolization was introduced in the treatment of patients with unresectable hepatocellular carcinoma waiting for liver transplantation. The rationale for this preoperative treatment was to control tumor growth during the waiting period and to improve long-term survival. This study aimed to investigate whether preoperative chemoembolization not only induces marked tumor necrosis but also has a survival benefit. METHODS: In this study 21 patients with hepatocellular carcinoma who underwent pretransplant chemoembolization (group I) were compared with 21 historical control patients (group II) without preoperative chemoembolization in a case-control study. The number of pretransplant chemoembolizations in each patient in group I varied between 1 and 5 with a mean of 2.44+/-1.15. In addition, six patients of this group received preoperative systemic chemotherapy. RESULTS: Overall, there were no differences in survival between the groups with and without pretransplant chemoembolization at 1 year (60.8% vs 61.5%) and at 3 years (48.4% vs 53.9%). In group I, three patients developed unexplained severe pneumonia, leading to death very early after liver transplantation. Marked tumor necrosis (>50%) was found in 14 cases in group I. In 6 out of these 14 patients, total tumor necrosis was observed. CONCLUSION: Although preoperative chemoembolization or chemotherapy induced marked tumor necrosis, these patients showed no benefit in survival compared to historical controls, and appeared to be at higher risk of developing immediate postoperative infective complications. PMID- 9875643 TI - Encapsulated xenogeneic hepatocytes remain functional after peritoneal implantation despite immunization of the host. AB - BACKGROUND/AIMS: Xenogeneic hepatocytes encapsulated in semipermeable membranes could be used in the future for the treatment of acute liver failure and congenital liver defects. However, host immune response could affect the viability and function of transplanted cells. The purpose of this study was to investigate the immunological consequences of intraperitoneal implantation of encapsulated xenogeneic hepatocytes and their effects. METHODS: Recipient Lewis rats received 2 x 10(7) human hepatocytes encapsulated in semipermeable hydrogel based hollow fibers, 2 x 10(7) free human hepatocytes or 2 x 10(7) encapsulated Lewis rat hepatocytes. The presence of human albumin in rat sera was assessed by Western blot and the presence of anti-human hepatocytes and anti-human albumin antibodies by ELISA. RESULTS: Anti-hepatocyte antibodies were detected on the 7th day, and their level increased progressively on days 21 and 28 in rats grafted with encapsulated or free human hepatocytes. Anti-albumin antibodies were detected on day 7 and increased progressively in rats grafted with encapsulated human hepatocytes, but were not detected in the other groups. No immune complexes or complement components of donor origin were detected by immunofluorescence in the recipients' tissues. Despite immunization of the host, encapsulated xenogeneic hepatocytes survived and produced albumin, whereas free hepatocytes had been lysed. CONCLUSION: Transplantation of encapsulated xenogeneic hepatocytes resulted in immunization of the host with production of anti hepatocyte and anti-albumin antibodies. However, hepatocytes could be efficiently protected by the membrane and remained viable and functional during the study. PMID- 9875644 TI - Protective effect of prostaglandin E1 against ischemia/reperfusion-induced liver injury: results of a prospective, randomized study in cirrhotic patients undergoing subsegmentectomy. AB - BACKGROUND/AIMS: The cytoprotective effects of prostaglandin E1 on livers suffering from ischemia/reperfusion injury in the clinical setting are unproved. These effects were examined, focusing on inflammatory cytokine and nitric oxide metabolism. METHODS: Twenty-four cirrhotic patients with hepatocellular carcinoma undergoing subsegmentectomy under ischemia induced only by Pringle's maneuver were divided into two groups (patients given prostaglandin E1 by injection and untreated controls) and postoperative results were compared. Peripheral blood was taken perioperatively and the plasma aminotransferase, cytokines and nitrate/nitrite levels of the two groups were compared. Two liver specimens were taken from each patient, one before ischemia and the other after hepatectomy, and the levels of inducible nitric oxide synthase and cytokine mRNAs and proteins were analyzed. RESULTS: Although no apparent differences were recognized in postoperative complications or duration of postoperative hospital stay between the groups, the perioperative plasma aminotransferase level was significantly lower in the prostaglandin E1 group. Significant differences were also seen in interleukin-6 and nitrate plasma levels during the observation period and the interleukin-6 protein levels in the liver supernatants after hepatectomy in the two groups. In contrast, no significant differences were apparent between the interleukin-1 beta and tumor necrosis factor-alpha plasma levels of the two groups. The corrected fluorescence activities of interleukin-6 and inducible nitric oxide synthase mRNAs in the liver after hepatectomy correlated significantly. No interleukin-1 beta or tumor necrosis factor-alpha mRNAs or proteins were detected. CONCLUSIONS: Prostaglandin E1 exerted hepatoprotective effects on livers suffering from ischemia/reperfusion injury, and interleukin-6 might play an important role in these effects. PMID- 9875645 TI - Immediate onset of DNA synthesis in remnant rat liver after 90% hepatectomy by an administration of follistatin. AB - BACKGROUND/AIM: Follistatin is an antagonist of activins and is effective in promoting liver regeneration after 70% hepatectomy. To examine its efficacy under more critical conditions, we studied the effect of follistatin on liver regeneration in 90% hepatectomized rat. METHODS: Human recombinant follistatin was infused into the portal vein immediately after 90%, hepatectomy in 24-h starved rats, and changes in the liver regeneration rate and nuclear bromodeoxyuridine labeling were measured. RESULTS: In control rats, nuclear labeling was first detected at 11 h of hepatectomy. In follistatin-treated rats, nuclear labeling was markedly increased at 3 h, and was significantly higher than that in control rats at 24, 72, 96, 120 and 144 h. The liver regeneration rate was significantly higher in follistatin-treated rats at 48, 72, 96, 120, 144 and 168 h. To determine the reason for the accelerated growth in starved rats, we compared the expression of mRNA for c-myc, p53, p21CIP1, p15INK4B, p27KIP1, and subunits of activins in fed and starved rats. mRNA for p21CIP1 and p15INK4B, but not p27KIP1 were decreased in 24 h-starved rats compared to the fed rats. mRNA for betaA subunit of activin was not detected in either fed or 24-h-starved rats, whereas that for betaC subunit was increased in starved rats. CONCLUSION: These results indicate that follistatin induces immediate onset of DNA synthesis in 90% hepatectomized rats and is quite effective in promoting liver regeneration. PMID- 9875646 TI - Lamivudine treatment for acute hepatitis B after liver transplantation. AB - BACKGROUND/AIMS: Acute hepatitis caused by recurrent or de novo hepatitis B virus (HBV) infection after liver transplantation frequently induces aggressive disease leading to liver failure. The aim of this study was to determine the efficacy and safety of lamivudine treatment in post-transplant acute hepatitis B. METHOD: Twelve patients with acute hepatitis B were started on lamivudine 100 mg p.o. daily within 8 weeks of the appearance of HBsAg. One patient was excluded after 1 month because of hepatocellular carcinoma recurrence. Patients were followed for an average of 68.6 weeks (range 32-108), and were clinically and biochemically evaluated on a monthly basis. They had a histological assessment at baseline, after at least 6 months, and whenever clinically indicated. RESULTS: Basal HBV DNA ranged between 13 and 1288 pg/ml and serum alanine aminotransferase between 97 and 1036 U/l. HBV-DNA became undetectable within 8 weeks and transaminases normalized within 24 weeks in all cases. At the last visit, eight patients (73%) remained HBV-DNA negative by liquid hybridization and had normal or close to normal alanine aminotransferase. Five patients (45%) were also HBsAg negative and HBV-DNA negative by polymerase chain reaction. HBV-DNA and transaminase breakthrough occurred in three patients (27%). Histology after 6-9 months showed chronic hepatitis in seven patients. Lamivudine was well tolerated without serious adverse reactions. CONCLUSIONS: These results indicate that lamivudine treatment induces sustained inhibition of viral replication and normalization of transaminases in the majority of post-transplant patients with acute hepatitis B. HBsAg loss may be achieved in a considerable number of cases. Although viral resistance is relatively frequent, early initiation of lamivudine appears to be effective and safe. PMID- 9875647 TI - Successful treatment of refractory type 1 autoimmune hepatitis with methotrexate. AB - BACKGROUND/AIMS: Autoimmune hepatitis is a heterogeneous disorder that typically responds to glucocorticoids with or without azathioprine. Treatment options for patients not responding to standard therapy are limited. METHODS: We describe a 52-year-old female who presented with jaundice, marked elevation in liver enzymes, positive antinuclear antibody and a liver biopsy consistent with autoimmune hepatitis. Liver enzymes did not normalize with prednisone alone. When azathioprine was added, the disease flared. The patient refused cyclosporine. Methotrexate 7.5 mg po per week resulted in normalization of liver enzymes, improved liver histology, and has maintained remission with a steroid-sparing effect. RESULTS/CONCLUSION: In this patient methotrexate was used successfully to treat type 1 autoimmune hepatitis. This suggests that methotrexate may have a role in treatment of autoimmune hepatitis refractory to standard therapy. PMID- 9875648 TI - Long-term mebendazole therapy may be parasitocidal in alveolar echinococcosis. AB - Long-term chemotherapy of human alveolar echinococcosis with benzimidazole compounds (mebendazole, albendazole) has been shown to be primarily parasitostatic, but its curative (parasitocidal) efficacy is debated. This article reports on a 67-year-old male patient with non-resectable alveolar echinococcosis of the liver who had been continuously treated for 13 years with mebendazole (approximately 45-48 mg/kg body weight per day) and who was closely monitored according to a specific protocol. At the age of 80 years the patient died of oesophageal variceal bleeding. During treatment the hepatic lesion had decreased markedly in size in association with progression of perifocal calcification. At autopsy, a well-demarcated, necrotic, partially calcified, parasite-induced lesion of the right liver lobe and secondary biliary cirrhosis were found. Remnants of parasite tissue obtained from the periphery of the lesion showed a small-cystic structure, but it was not viable, as evidenced by transplantation of tissue blocks to rodents. The case is suggestive for a parasitocidal efficacy of mebendazole treatment carried out for 13 years, and is discussed in context with conflicting literature data. PMID- 9875649 TI - Graft loss and the antiphospholipid syndrome following liver transplantation. AB - BACKGROUND/AIMS: The antiphospholipid syndrome is characterised by arterial and venous thromboses affecting small and large vessels, together with the presence of lupus anticoagulant and anticardiolipin antibodies. METHODS: We report two cases of the antiphospholipid syndrome following liver transplantation which resulted in hepatic vessel thrombosis and subsequent graft loss, and we discuss the value of anticoagulation in prevention of further thrombotic episodes. Identification of these cases prompted a retrospective analysis for the presence of IgG anticardiolipin antibody, the most useful laboratory marker of vascular thrombosis in the antiphospholipid syndrome. Serum taken at 2 to 4 weeks following transplantation (when thrombotic events might be anticipated) was analysed further in 132 patients undergoing liver transplantation. RESULTS: Hepatic vessel thrombosis occurred in 21/132 (16%) post transplantation. Anticardiolipin IgG was present in 2/21 (8.8%) recipients who developed a hepatic vessel thrombosis following liver transplantation, compared to 7/111 (6.3%) (p=0.59) in whom transplantation was not complicated by hepatic vessel thrombosis. CONCLUSIONS: The presence of IgG anticardiolipin antibody was uncommon in recipients following liver transplantation and was not associated with an increased risk of hepatic vessel thrombosis. In contrast, the antiphospholipid syndrome which is rare may result in hepatic artery occlusion and graft loss, and so prompt recognition of the clinical syndrome is critical. Although screening for the antibody is not warranted in liver transplant recipients, it is important to be aware of the syndrome as optimal management of such cases may prevent graft loss. PMID- 9875650 TI - Images in hepatology. Mediastinal mass in a patient with cirrhosis. PMID- 9875651 TI - Focal nodular hyperplasia in a young female. AB - A 41-year-old woman, who was taking oral contraceptive drugs, presented with a symptomless hepatic mass lesion shown by abdominal ultrasound. Doppler ultrasound, computed tomography (unenhanced and enhanced with contrast) and magnetic resonance imaging after intravenous gadolinium suggested this was focal nodular hyperplasia. Ultrasound-guided liver biopsy had been performed elsewhere and was also diagnostic. The differential diagnosis of hepatic mass lesion is illustrated and particularly the distinction of focal nodular hyperplasia from adenoma. The importance of radiological imaging techniques is stressed. Clinical management is discussed. PMID- 9875652 TI - Combination therapy of chronic hepatitis C: an important step but not the final goal! PMID- 9875653 TI - Histological grading and staging in chronic hepatitis: clinical applications and problems. PMID- 9875654 TI - Hepatitis C in the patient with human immunodeficiency virus infection. PMID- 9875655 TI - Protease inhibitors suppress TGF-beta generation by hepatic stellate cells. PMID- 9875656 TI - Coupling to lactosaminated poly-L-lysine reduces the toxic effects of ribavirin on red blood cells. PMID- 9875657 TI - Severe anemia following combined alpha-interferon/ribavirin therapy of chronic hepatitis C. PMID- 9875658 TI - Hepatitis C virus (HCV)-infection and oral lichen planus. PMID- 9875659 TI - Danazol therapy: an unusual aetiology of hepatocellular carcinoma. PMID- 9875660 TI - Phenotype variability of the dominant beta-thalassemia induced in four Dutch families by the rare cd121 (G-->T) mutation. AB - Eight patients who were carriers of beta-thalassemia induced by the cd121 (G-->T) mutation are described in four nonrelated Dutch families. This mutant, which is considered rare and inherited in a dominant manner, is expressed in a different way among each of the four families and even among carriers of the same family. The symptoms vary from an hemolytic anemia of intermediate gravity with hepatosplenomegaly, inclusion bodies and erythroblastosis, to a mild anemia with minor hematological abnormalities. We report the analytical procedures used for the detection of the mutant, the hematological and clinical data of the four families and discuss the variable physiopathology of this molecular defect. We also compare the variation in fetal hemoglobin expression in relation to the haplotypes of the beta-gene cluster and to the different hematological conditions. The presence of this rare mutant in four nonrelated Dutch families could derive from a single mutation or from multiple events. The existence of the four mutations in three different haplotypes suggests the occurrence of at least two independent events. The presence of five abnormal hemoglobins and the beta thalassemia defect on different haplotypes at cd121 also suggests a relatively increased rate of mutations at this particular site. PMID- 9875661 TI - Prevalence and molecular characterization of beta-thalassemia in Filipinos. AB - Beta-thalassemia (thal) is a common single-gene disease worldwide. However, the prevalence of beta-thal and the spectrum of beta-globin gene mutations in Filipinos remain unclear. This study sought to answer these two questions. A total of 2954 apparently healthy Filipinos in Taiwan were recruited for a prevalence study. A complete blood count was done in every subject. Those with microcytosis were studied with hemoglobin (Hb) high-performance liquid chromatography to determine the levels of Hb A2 and Hb F. Twenty-seven subjects had elevated levels of Hb A2 (>4.0%). These 27 suspected beta-thal carriers and another 16 beta-thal major patients who were being treated in the Philippines were studied to determine the spectrum of beta-globin gene mutations. Gap-PCR was used to detect the Filipino deletion of beta-thal, and direct sequencing was used to detect point or small mutations in the beta-globin gene. All of the 27 suspected beta-thal carriers had one mutation in the beta-globin gene, resulting in an overall prevalence of 0.9%. The spectrum of beta-thal mutations was similar in the carrier and patient groups. Analysis of the pooled identified seven different mutations in the study population. The Filipino deletion was the most common mutation, accounting for 45.8% (27/59) of the alleles, followed by codon 67 (-TG) (16 alleles), and Hb E (11 alleles). These three mutations accounted for 92% of the Filipino beta-thal alleles. Elucidation of the beta-thal mutations in Filipinos is useful for the genetic counseling and prenatal diagnosis of this disease. PMID- 9875662 TI - Significance of p53 overexpression in bone marrow biopsies from patients with bone marrow failure: aplastic anemia, hypocellular refractory anemia, and hypercellular refractory anemia. AB - Among patients with bone marrow failure, differentiating acquired aplastic anemia (AA) from hypocellular refractory anemia (hypo RA) can be a difficult and challenging task. Morphological, cytochemical, immunocytochemical, and cytogenetic studies may provide tools for discriminating between both entities. In addition, differences in the pattern of proliferation and apoptosis of bone marrow cells in AA and in the myelodysplastic syndrome have been reported. Because of the correlation between p53 and apoptosis, we examined the overexpression of p53 on bone marrow biopsies in RA and AA. Our study included 14 patients with hypo RA, 14 patients with hypercellular (hyper) RA, ten patients with classic acquired AA, and 37 hematologically normal individuals. p53 was overexpressed in eight (57%) hypo RA patients and 11 (79%) hyper RA patients. All normal individuals and patients with AA showed no overexpression of p53 in their marrow. These results were statistically significant:p < 0.01 (AA vs hypo RA), p<0.001 (AA vs hyper RA), while the difference between hypo RA and hyper RA was not statistically significant. We conclude that p53 overexpression in bone marrow biopsies is a valuable tool for studying bone marrow failure and may provide additional information to help differentiate hypo RA from acquired AA. PMID- 9875663 TI - Different binding mechanisms of myeloid leukemic cells to adhesion molecules on bone marrow stromal fibroblasts induced by TNF-alpha and IL-4. AB - To study the mechanisms of adhesion of myeloid leukemic cells to bone marrow stroma, we analyzed the interaction of bone marrow stromal fibroblasts with myeloid leukemic cell lines and the modulation of adhesion molecule expression on stromal fibroblasts by TNF-alpha and IL-4. Like others, we found up-regulation of VCAM-1 and ICAM-1 on fibroblasts with TNF-alpha treatment, whereby IL-4 acted synergistically with TNF-alpha. VCAM-1 expression on the cell surface was maximal after 10 h, while ICAM-1 expression increased up to 48 h. All myeloid leukemic cell lines tested (HL-60, K562, TMM, U937, ML-2, PLB-985, THP-1, KG1a) revealed weak adhesion to untreated bone marrow fibroblasts (< or =10% bound cells). TNF alpha and IL-4 significantly enhanced adhesiveness of fibroblasts to the cell lines PLB-985, THP-1, and ML-2, with a peak between 6 and 10 h of treatment. Adhesiveness to the cell line TMM was increased up to eightfold in a time dependent manner for up to 48 h. The enhanced binding of ML-2-, THP-1-, and PLB 985 cells to stimulated fibroblasts was due at least partially to the interaction of VLA-4 with VCAM-1. Increased adhesion of TMM cells was impaired neither by antibodies to VLA-4, LFA-1, or Mac-1 nor by antibodies to their counter-receptors VCAM-1 or ICAM-1, suggesting that adhesion molecules distinct from VCAM-1 or ICAM 1 are involved in enhanced adhesiveness of the fibroblasts to myeloid leukemic cells. PMID- 9875665 TI - Hypertrophic cranial pachymeningitis in a patient with aplastic anemia. AB - We report on a 13-year old girl with severe aplastic anemia and hypertrophic cranial pachymeningitis. She was admitted to our hospital with severe headache and vomiting. A computerized tomographic (CT) scan of the brain on the third day of symptoms showed a hyperdense area in the tentorial region. Magnetic resonance imaging (MRI) showed iso-intensity in the same tentorial region in T1- and T2 weighted images, and gadolinium enhancement of this region suggested a thickened dura mater. Initially, a diagnosis of subdural or subarachnoid hemorrhage was made. Since her platelet count was low (3000/microl) making the patient a poor risk candidate for surgery, and the area was limited to the dura mater, conservative therapy, including glycerol administration and platelet transfusion, was carried out. Despite clinical improvement 10 days after admission without specific therapy, the iso-intense region on the left side of the tentorial region remained unchanged on MRI. On the other hand, the iso-intense area on the right side of the tentorial region became hyperdense on T1-weighted MRI images and was also enhanced by gadolinium. Cerebrospinal fluid findings were normal except for slightly elevated protein at 62 mg/dl. A diagnosis of hypertrophic cranial pachymeningitis of the tentorial dura mater with hemorrhage on the right side was made. Although hypertrophic cranial pachymeningitis is a rare disease, it must be considered in the differential diagnosis of severe headache in a case of aplastic anemia. PMID- 9875664 TI - Absence of a directly causative role for human herpesvirus 7 in human lymphoma and a review of human herpesvirus 6 in human malignancy. AB - In search of a (new) viral etiological agent, we screened 64 lymph node samples from Hodgkin's disease (HD) and 43 samples (32 lymph node and 11 skin biopsies) from non-Hodgkin's lymphoma (NHL) for human herpesvirus 7 (HHV-7). Twenty-nine control samples were tested as well, including 17 with benign lymphadenopathy. None of the samples tested positive by Southern blot hybridization using HHV-7 specific probes. We conclude that there is no major HHV-7 load in human lymphoma and that HHV-7 is not likely to be directly involved in its etiology. This is in contrast to a small minority of human lymphoproliferative diseases in which HHV-6 can be found at high copy number, but in which an etiological role is still uncertain. PMID- 9875666 TI - Graft-versus-host disease following second syngeneic stem cell transplantation for relapsed chronic myeloid leukemia. AB - Allogeneic bone marrow transplantation is the only curative treatment for patients with Philadelphia chromosome-positive chronic myeloid leukemia (CML); however, recurrence of disease remains a major cause of treatment failure. A 26 year-old man with chronic myeloid leukemia who had a cytogenetic relapse 49 months after his first syngeneic bone marrow transplant (BMT) and hematologic relapse 23 months thereafter progressed to blast crisis despite treatment with IFN-alpha for 15 months. He underwent a second transplantation in early second blast crisis, 92 months after the first BMT with PBPC from his previous donor. Successful hematological reconstitution occurred. On day 50 after the second transplantation the patient developed a generalized rash, hepatomegaly, and cholestatic signs. Skin and liver biopsy revealed changes compatible with acute graft-versus-host disease (GVHD). Treatment with cyclosporin A (CSA) and prednisone was started, and the GVHD resolved. Fifteen months after PBPC transplantation he had a molecular relapse. Despite discontinuation of CSA, the patient progressed into blast crisis 7 months later. The occurrence of GVHD and disappearance of the BCR-ABL-positive clone suggest that a graft-versus-leukemia (GVL) effect may have been operative for 15 months in a patient given a second syngeneic BMT in blast crisis. PMID- 9875667 TI - Anaplastic large-cell lymphoma of null-cell type with multiple bone involvement. AB - A 21-year-old man who had anaplastic large cell lymphoma (ALCL) of the null-cell type with multiple bone involvement is reported. On admission, he had symptoms of incomplete paraplegia and urinary and rectal incontinence. Workup studies for staging revealed para-aortic lymph node swellings and multiple bone involvement including skull, ribs, left iliac bone, and thoracic/lumbar spine. Because paraplegia was rapidly progressive, a decompression operation was performed. The biopsy specimen obtained from the lumbar spine revealed sheetlike proliferation of anaplastic large cells. These cells were positive for CD30 (Ki-1), EMA, vimentin, and p80NPM/ALK, and negative for CD3, CD20 (L26), and CD45 (LCA). Epstein-Barr virus-encoded small RNAs were not detectable in these cells. Thus, the patient was diagnosed as having ALCL of the null-cell type. He was treated with several courses of combination chemotherapy, and finally with total body irradiation plus high-dose chemotherapy supported by peripheral blood stem cell transplantation. However, soon after the treatment, the lymphoma cells massively infiltrated his bone marrow. He died of lymphoma 8 months after admission. PMID- 9875668 TI - Treatment of essential thrombocythemia during pregnancy: antiabortive effect of interferon-alpha? PMID- 9875669 TI - Pharmacokinetic study of busulfan in an AML patient treated with regular hemodialysis. PMID- 9875670 TI - Genetic immunization against neu/erbB2 transgenic breast cancer. AB - erbB2/neu, an overexpressed oncogene product, has been proposed as a human cancer vaccine target. In the present study, transgenic (rat neuNT oncogene) FVB/neu mice, developing metastasizable mammary carcinoma, were immunized with a plasmid DNA encoding are not tolerant to the self antigen and sequences. We report that transgenic tumour-bearing mice, like some breast cancer patients erbB2 + X, develop anti-neu autoimmune responses, which can be boosted and skewed to a Th1 phenotype by DNA immunization. Intramuscular injections of neuNT plasmid drastically reduced (or even prevented in a small number of treated mice) the outgrowth of mammary neoplasms as well as their metastatic penetrance. Furthermore, DNA immunization caused haemorrhagic necrosis of established cancer nests, leaving a greatly reduced portion of the tumour burden for the host to cope with. The antitumour activities we obtained, in this very challenging model for cancer immunotherapy, lay the foundation for DNA-based immunization to control erbB2/neu-overexpressing neoplasms. PMID- 9875671 TI - Spontaneous T cell responses to melanoma differentiation antigens from melanoma patients and healthy subjects. AB - The spontaneous cytotoxic T cell responses to melanoma differentiation antigens and influenza matrix peptide were compared in 20 HLA-A2+ melanoma patients and 17 healthy A2+ individuals. Cytotoxic T lymphocyte (CTL) responses were determined by mixed lymphocyte peptide culture (MLPC) involving two stimulations of unfractionated peripheral blood lymphocytes (PBLs) with peptide in vitro. CTL responses to Melan-A 9-mer (amino acids 27-35, AAGIGILTV) peptide were detected in 4 out of 16 normal individuals, but in none of the melanoma patients. CTL specific for influenza matrix peptide were frequently found in both normal individuals and melanoma patients, suggesting that generalized immuno-suppression was not responsible for this difference. No significant responses were observed in either normal individuals or melanoma patients to Melan-A 10-mer (26-35, EAAGIGILTV), two gp1OO epitopes (280-288, YLEPGPVTA; 457466, LLDGTATLRL) and two tyrosinase epitopes (1-9, MLLAVLYCL; 368-376, YMDGMSQV). Melan-A (27-35)-specific CTL cells generated by normal individuals and melanoma patients recognized both synthetic peptide-pulsed T2 cells and two HLA-A2+, Melan-A+ melanoma cell lines (ME272, LAR1) in an antigen-specific, MHC class I restricted manner. T cells generated against Melan-A 9-mer were also able to recognize Melan-A 10-mer-pulsed target cells. Spontaneous CTL responses to Melan-A 9-mer from three known responder normal individuals were further evaluated over a prolonged time course (6-11 months). All 3 subjects demonstrated specific Melan-A 9-mer responses throughout the study period, although lytic activity fluctuated over time for a given individual. We found the MLPC assay to be reliable and easy to perform for monitoring T cell responses, although it may still not be sufficiently sensitive to detect low numbers of precursor T cells. PMID- 9875672 TI - Characterization of B16 melanoma-specific cytotoxic T lymphocytes. AB - A B16 melanoma-specific CD8+ T cell line (AB1) was established from the spleen cells of C57BL/6 mice cured of B16 melanoma with interleukin (IL)-12 treatment. The AB1 line exclusively used T cell receptor Vbeta11. The AB1 cells exhibited a cytolytic activity against both syngeneic B16 melanoma and allogeneic P815 mastocytoma, whereas a cold inhibition assay revealed specificity of the AB1 cells against B16 melanoma. Their lostability to kill a class I loss variant of B16 melanoma was restored by the transfection of H-2Kb gene. In addition, their interferon (IFN)-gamma production was significantly suppressed by the addition of anti-H-2Kb monoclonal antibody, and RT-PCR analysis showed that the AB1 line expressed the mRNA encoding IFN-gamma, but not IL-4 or IL-10. The experiment using synthetic peptides of tyrosinase-related protein-2 (TRP-2) revealed that the AB1 cells could recognize TRP-2(181-188) peptide. Moreover, the AB1 cells showed an in vivo antitumor effect against established pulmonary metastases of B16 melanoma. Overall, these results indicate that the Tc1-type Vbeta11+ AB1 cells exert an antitumor activity against syngeneic B16 melanoma through recognition of TRP-2(181-188) peptide in an H-2Kb-restricted manner. PMID- 9875673 TI - Milky spots in the greater omentum are predominant sites of local tumour cell proliferation and accumulation in the peritoneal cavity. AB - The role that milky spots in the greater omentum play in tumour cell spread in the peritoneal cavity is presently not fully understood. To study whether intraperitoneally injected tumour cells appear preferentially in milky spots of the greater omentum and to study the changes in the greater omentum, and especially in the cell population of milky spots after tumour cell infiltration, the following study was performed. A detailed temporal sequences of changes in morphology and cellular composition in milky spots of the greater omentum of Wag/Rij rats 5, 15, 30, 60 min, 2, 4, 8, 16, 24 h, 2, 4, 8 days and 2 and 4 weeks after intraperitoneal administration of 2.0 x 10(6) CC 531 tumour cells was investigated by light microscopy and electron microscopy (pre-embedding labelling). Our data showed that the milky spots in the greater omentum were the sites to which tumour cells migrated preferentially from the peritoneal cavity. The tumour cells infiltrated the milky spots and formed clusters within. The cellular population in milky spots reacted by a very rapid influx of young macrophages during the first hour and an increase of the total number of cells (P < 0.01). After 4 h tumour cells were also located on the greater omentum outside the area of the milky spots. Around these tumour cell deposits, new milky spots are formed, which increased the total number of milky spots. The cells present in milky spots are not capable of reversing the growth of tumours and finally a solid omental cake of tumour cells is formed. PMID- 9875674 TI - Antitumor response of regional lymph node lymphocytes in human lung cancer. AB - We recently reported that regional lymph node lymphocytes (RLNL) from patients with primary lung cancer were in a more highly activated state than peripheral blood lymphocytes (PBL) when the activation-related molecules were studied by FACS analysis. To identify whether or not RLNL had the ability to respond to autologous tumor cells (AT), in the present study a mixed lymphocyte tumor cell reaction (MLTR) either with or without recombinant interleukin 2 (rlL-2) was performed in 41 cases with primary lung cancer. Significant proliferative responses to AT were found in RLNL from 20 of 41 cases (48.8%) without IL-2, and in 23 of 41 cases (56.1%) with IL-2. On the other hand, such responses were observed in the PBL from 8 of 30 cases (26.7%) without IL-2, and from 11 of 30 cases (36.7%) with IL-2. No significant correlation between MLTR and such clinical factors as tumor size, metastasis to lymph node, histology and stage of the disease was found. To further analyze the anti-AT response of RLNL, the cytokine production of RLNL was investigated after stimulation by AT. An increase in interferon-gamma (IFN-gamma) production was observed in RLNL with a positive reaction of MLTR, while no such increase was found in PBL. Finally, to elucidate whether the expression of MHC class II molecules was a key point in MLTR, tumor cells in primary lesions were examined for the expression of MHC class II by immunohistochemical staining, and the blocking assay of MLTR was performed with anti-MHC class II monoclonal antibody. Data suggested that there was a positive correlation between MHC class II expression of the tumor and MLTR and that MLTR were partially blocked by anti-MHC class II monoclonal antibody. These results demonstrated that RLNL were in a more highly activated state against AT than were PBL, and this finding is considered to be helpful in enhancing our understanding of the role of RLNL in lung cancer patients. PMID- 9875675 TI - Generation of cytokine-induced killer cells using exogenous interleukin-2, -7 or 12. AB - Immunologic effector cells termed cytokine-induced killer (CIK) cells are generated in vitro from peripheral blood lymphocytes by addition of interferon gamma, interleukin (IL)-2, IL-1 and an antibody against CD3. CIK cells have been shown to eradicate established tumors in a SCID mouse/human lymphoma model. CIK cells are dependent on exogenous cytokines such as IL-2, IL-7, or IL-12. We studied the effect of these cytokines in detail. Cellular proliferation was analyzed using an MTT proliferation assay, surface antigen expression via flow cytometry, cytotoxic activity using an LDH release assay, and apoptosis via flow cytometric analysis. IL-2, IL-7 and IL-12 led to significant growth of lymphocytes. Cells grown in IL-2 and IL-7 showed higher proliferation rates than cells grown in IL-12 according to the MTT assay. Concerning surface antigen expression, exogenous IL-7 led to a decrease in IL-7 receptor expression (4.8% from 60.4%) and exogenous IL-2 to a decrease in IL-2 receptor expression (61.2% from 73.2%). CD28 expression was higher in cells grown in IL-7 (77.3%) than in cells grown in IL-2 (62.5%). IL-12 led to a decrease in ICAM-1 adhesion molecule expression (57.7% from 76.7%) and an increase in CD56 expression compared with exogenous IL-7. IL-7 led to higher number of CD4-positive cells than IL-2 (53.0% vs 49.5%). No significant difference was found between IL-2, IL-7 and IL-12 in cytotoxic activity measured in an LDH release assay. Small amounts of apoptotic cells were found with all cytokines. However, the percentage of necrotic cells was higher with exogenous IL-12 than with IL-2 or IL-7. In summary, CIK cells can be generated using exogenous IL-2, IL-7 or IL-12. No difference in cytotoxic activity was found. However, significant differences were found in cell proliferation rates, antigen expression and percentage of necrotic cells. PMID- 9875676 TI - Effect of human interferon beta gene transfer upon human glioma, transplanted into nude mouse brain, involves induced natural killer cells. AB - We investigated the immunological responses induced by human interferon beta (IFNbeta) gene transfer in human gliomas produced in the brains of nude mice. A suspension of human glioma U251-SP cells was injected into the brains of nude mice. The IFNbeta gene was transferred by intratumoral injection with cationic liposomes or cationic liposomes associated with anti-glioma monoclonal antibody (immunoliposomes). When intratumoral injection of liposomes or immunoliposomes containing the human IFNbeta gene was performed every second day for a total of six injections, starting 7 days after tumor transplantation, complete disappearance of the tumor was observed in six of seven mice that had received liposomes and in all seven mice receiving immunoliposomes. In addition, experimental gliomas injected with immunoliposomes were much smaller than those injected with ordinary liposomes following delayed injections beginning 14 days after transplantation. An immunohistochemical study of the treated nude mouse brains revealed a remarkable induction of natural killer (NK) cells expressing asialoGM1 antigen. To investigate the significance of NK cells in the antitumor effect, we injected liposomes or immunoliposomes containing the human IFNbeta gene into tumors in nude mice depleted of NK cells by irradiation and anti asialoGM1 antibody administration. The antitumor effect of the liposomes or immunoliposomes was abolished. Subsequent subcutaneous glioma challenge of the nude mice after intracerebral tumor implantation and gene transfer resulted in no subcutaneous tumor growth. These results suggest that the induction of NK cells is important in the cytocidal effect of liposomes or immunoliposomes containing the human IFNbeta gene upon experimental gliomas. PMID- 9875677 TI - cis-Diamminedichloroplatinum and 5-fluorouracil are potent inducers of the cytokines and natural killer cell activity in vivo and in vitro. AB - It has been reported that certain chemotherapeutic agents exhibit effects that enhance the antitumor host responses in the patients with malignant diseases. In the present study, we investigated whether cis-diamminedichloroplatinum (cisplatin) and 5-fluorouracil (5-FU) may induce cytokines and effector cells with antitumor efficacy in vivo and in vitro. The cultivation of human peripheral blood mononuclear cells (PBMC) in the presence of cisplatin (0-1.0 microg/ml) or 5-FU (0-5.0 microg/ml) resulted in the significant augmentation of natural killer (NK) and lymphokine-activated killer (LAK) cell activities as well as generation of interferon (IFN) gamma, tumor necrosis factor (TNF) alpha, beta interleukin(IL)-1beta, IL-6 and IL-12 in vitro. In addition, all of these activities were almost completely neutralized by addition of anti-asialoGM1 antibody and complement (P < 0.05). In an in vivo model, the administration of anti-asialoGM1 antibody significantly shortened the survival time extended by the treatment with cisplatin or 5-FU (P < 0.05), both on nude mice bearing salivary gland tumors and on syngeneic MethA-tumor-bearing BALB/c mice. Furthermore, high levels of NK and LAK activities and significant increases of the numbers of cells positive for asialoGM1, IFNgamma, TNFalpha, or IL-1beta were detected in the spleen cells derived from animals given cisplatin or 5-FU as compared with those given saline (P < 0.001-0.05). These findings clearly indicate that cisplatin and 5-FU are potent inducers of several types of cytokines and effector cells carrying antitumor activity mediated by asialoGM1-positive cells (mainly NK cells) for the most part, and that these abilities are closely associated with the in vivo antitumor effect of these agents. PMID- 9875678 TI - Antioxidant therapy--a new therapeutic option for reducing mortality from coronary artery disease. PMID- 9875679 TI - Reflections on pain. PMID- 9875680 TI - Pharmacokinetic interactions between acute alcohol ingestion and single doses of benzodiazepines, and tricyclic and tetracyclic antidepressants -- an update. AB - Recent reports of interactions between alcohol and benzodiazepines, tricyclic and tetracyclic antidepressants during their acute concomitant use are reviewed. Acute ingestion of alcohol (ethanol) with tranquilizers or hypnotics is responsible for several pharmacokinetic interactions that can have significant clinical implications. In general, metabolism of these drugs is delayed when combined with alcohol but some reports have suggested otherwise. The amount of alcohol consumed, the presence or absence of liver disease, and differences in the dosage and administration of these drugs may account for the observed discrepancies. In recent years, the cytochrome P450 (P450 or CYP) isoenzyme that catalyses the metabolism of these drugs has also been identified. However, since changes in the pharmacogenetic metabolism of benzodiazepines and tricyclic and tetracyclic antidepressants are mainly governed by CYP2C19 and CYP2D6, caution is needed when used together with alcohol. PMID- 9875681 TI - Does cholesterol lowering prevent stroke? AB - The importance of lowering plasma cholesterol to reduce the incidence of coronary events is well established. However, in the prevention of stroke disease, control of hypertension has been the main aim of treatment and lipid lowering therapy has not hitherto been considered to be desirable or necessary. In this review, the evidence from large multicentre trials, imaging studies and meta-analyses is presented. It shows convincingly that HMG-CoA reductase inhibitors (Statins) reduce stroke risk. PMID- 9875682 TI - Meta-analysis about efficacy of anti-resorptive drugs in post-menopausal osteoporosis. AB - OBJECTIVE: The purpose of this study was to compare the effect of three groups of anti-resorptive drugs in post-menopausal osteoporosis. DATA SOURCES: We collected data covering the period between 1983 and 1995, by first using MEDLINE. References retrieved were scanned further to identify additional papers. STUDY SELECTION: Only randomized studies evaluating bone mass by means of dual-photon or dual energy densitometry over a period of 1 year were accepted. DATA EXTRACTION: Studies were arranged into three drug groups. We used densitometry results after 1 year in all treatment or control groups. Factors which might interfere with the results were recorded for subsequent separate analysis. DATA SYNTHESIS: The MEDLINE search identified almost 25,000 studies. On reading the abstracts, 275 trials appeared to be controlled trials and original copies were retrieved for detailed analysis. A total of 31 articles which satisfied the inclusion criteria were identified. The first meta-analysis included studies which compared oestrogens and placebo, and the global effect-size was 0.54 (95% CI 0.34, 0.73). The second meta-analysis compared calcitonins with placebo and produced an effect-size of 0.41 (95% CI 0.21, 0.61) The third analysis compared bisphosphonates and placebo and showed an effect-size of 0.87 (95% CI 0.68, 1.07). Only oestrogen dose affected the results found. CONCLUSIONS: Bisphosphonates had the greatest effect on bone mass in post-menopausal osteoporosis. PMID- 9875683 TI - Use of in vitro models of haemofiltration and haemodiafiltration to estimate dosage regimens for critically ill patients prescribed cefpirome. AB - BACKGROUND: The physico-chemical properties of cefpirome (low protein binding, high water solubility and low molecular weight) suggest that it may be lost readily from the extracorporeal circulation of intensive care unit patients during continuous renal replacement therapy. METHOD: In order to make informed dosage recommendations for patients receiving artificial renal support, cefpirome loss from human blood has been quantified using in vitro models of continuous haemofiltration and haemodiafiltration. Cefpirome clearance was measured using three membrane types at varying ultrafiltrate (UFR) and dialysis flow rates (Qd). RESULTS: During haemofiltration cefpirome was found to cross hollow fibre polyamide (PA) and polyacrylonitrile (PAN) membranes with equal efficiency. The mean sieving coefficients (S) of both PA and PAN membranes were consistently high (> 0.7) when two different ultrafiltration rates were used. Changing the ultrafiltration rate or membrane type had no significant effect on the sieving coefficient of cefpirome but did result in an increase in cefpirome filter clearance (Fcl). Using the haemodiafiltration model, cefpirome penetrated PAN membranes (flat plate AN69S) more efficiently than hollow fibre PA membranes (FH66D). In each case, increasing the dialysis flow rate reduced the S-value. However, although increasing Qd was associated with a greater Fcl of cefpirome when PAN membranes were employed, no such relationship was found for the PA hollow fibre membrane. CONCLUSION: The information generated can be used to estimate a dosing regimen for intensive care patients prescribed cefpirome and receiving continuous renal replacement therapy. PMID- 9875684 TI - Serum protein binding kinetics of phenytoin in monotherapy patients. AB - OBJECTIVES: To determine the binding characteristics of phenytoin to serum proteins in the Japanese population and to compare these with those reported by other investigators. METHOD: Serum samples examined in the study were obtained from 72 patients (35 males, 37 females) receiving phenytoin monotherapy. The patients' ages ranged from 1 to 73 years (1-15 years, 36 subjects; 16-44 years, 20 subjects; 45-64 years, 13 subjects; > or = 65 years, 3 subjects). RESULTS: The in vivo population binding parameters of phenytoin to serum proteins and theoretical minimal unbound serum phenytoin fraction (fu) were determined using the Scatchard equation. The association constant (K) was 0.020 1/micromol, while the total concentration of binding sites (n(Pt) was 556 micromol/l. The number of binding sites per albumin molecule (n) was 0.85, while binding ability (n.K) was 0.017 l/micromol. The fu was 0.083. The n.K is approximately 1.1 times higher in patients of Pospisil et al. (26) (i.e. 0.0191 l/micromol) than in all our patients. The association constant is approximately 1.1 times higher in our study than in the in vitro study of Monks et al. (23) (i.e. 0-0186 l/micromol), while n is similar between the two studies. The fu in our patients is similar to the unbound serum phenytoin fraction in adult patients receiving phenytoin therapy reported by Richens (2) (i.e. 0.1). CONCLUSION: Our results suggest that there may be small differences in the binding characteristics of phenytoin to serum proteins between Japanese and non-Japanese subjects. The unbound serum fraction of phenytoin in our patients with epilepsy can be assumed to be relatively constant in the therapeutic concentration range of phenytoin. PMID- 9875685 TI - Therapeutic drug monitoring of tobramycin: once-daily versus twice-daily dosage schedules. AB - OBJECTIVE: To evaluate the effect of dosage regimen (once-daily vs. twice-daily) of tobramicyn on steady-state serum concentrations and toxicity. MATERIALS AND METHODS: Patients undergoing treatment with i.v. tobramycin (4 mg/kg/day) were randomised to two groups. Group OD (n = 22) received a once-daily dose of tobramycin and group TD (n = 21) received the same dose divided into two doses daily. Tobramycin serum concentrations (peak and trough) were measured by enzyme multiplied immunoassay. The renal and auditory functions of the patients were monitored before, during and immediately after treatment. RESULTS: The two groups were comparable with respect to sex, age, body weight and renal function. No statistically significant differences were found in mean daily dose, duration of treatment, or cumulative dose. Trough concentrations were < 2 g/ml in the two groups (100%). Peak concentrations were > 6 microg/ml in 100% of the OD group and in 67% of the TD group (P< 0.01). Mean peak concentrations were markedly different: 11.00+/-2.89 microg/ml in OD vs. 6.53+/-1.45 microg/ml in TD (P< 0.01). The pharmacokinetics parameters were: Ke, (0.15+/-0.03/h in OD vs. 0.24+/ 0.06/h in TD), t1/2, (4.95+/-1.41 h in OD vs. 3.07+/-0.71 h in TD), Vd (0.35+/ 0.11 l/kg in OD vs. 0.33+/-0.09 l/kg in TD), Cl (0.86+/-0.29 ml/min/kg in OD vs. 1.28+/-0.33 ml/min/kg in TD). Increased serum creatinine was observed in 73% of patients in OD versus 57% of patients in TD, without evidence of nephrotoxicity. In TD group, three patients developed decreased auditory function, of which one presented with an auditory loss of -30 dB, whereas in the OD group only one patient presented decreased auditory function. CONCLUSION: This small study suggests that a once-daily dosing regimen of tobramycin is at least as effective as and is no more and possibly less toxic than the twice-daily regimen. Using a single-dose therapy, peak concentration determination is not necessary, only trough samples should be monitored to ensure levels below 2 microg/ml. PMID- 9875686 TI - Study of the pharmacokinetics of cefpirome sulphate in the elderly. AB - OBJECTIVE: To determine the appropriate method of administration of the cephem antibiotic cefpirome sulphate in elderly patients. METHOD: We studied cefpirome's pharmacokinetics in patients with urinary tract infections. Patients received cefpirome sulphate 0.5 g by intravenous drip infusion over 30 mins. RESULTS: Patients with a creatinine clearance rate (Ccr) of 80 ml/min had an AUC of 96.7 microg.h/ml and a T1/2 of 2.36 h, whereas those with Ccr of 40-80 ml/min had an AUC of 172.0 microg.h/ml and a T1/2 of 3.45 h and those with Ccr of < 40 ml/min had an AUC of 152 microg.h/ ml and a T1/2 of 4.86 h. CONCLUSION: These results indicate that decreased kidney function can cause increases in the AUC and T1/2 of cefpirome. Thus in elderly patients and perhaps also in other patients with decreased kidney function, cefpirome should be administered at an initial dose of 0.5 g. PMID- 9875687 TI - Determination of heparin in aqueous solutions. AB - OBJECTIVE: To develop a volumetric method for assaying heparin in aqueous media. METHOD: Heparin is precipitated out with an aqueous solution of an organic amine by titration and the end-point is based on the measurement of the medium dielectric permittivity. We studied the titration of a 500 IU/ml heparin solution with a 0.08 M cetylpyridinium chloride solution at pH 6.8. Then, we assayed sulphate groups selectively at pH 2. The results were compared against a classical method of SO2 determination. The sensitivity and reproducibility of the volumetric method were evaluated at pH 6.8 and compared with the characteristics of a chromogenic method, usually used for the assay of heparin in biological fluids. RESULTS: A linear relationship between anticoagulant activity and sulphate and carboxyl group concentration was observed. The method was less sensitive but more reproducible than the chromogenic method. CONCLUSION: The proposed method can be used for aqueous solutions and is easy to carry out. It can be fully automated and applied to formulation studies. PMID- 9875689 TI - Quality of life measures in psoriasis: a critical appraisal of their quality. AB - In recent years, the importance of patient-generated evaluations in assessing the impact of healthcare has been recognized. A plethora of quality of life (QoL) instruments have been developed to measure the burden of psoriasis and its treatment on the patient. We review the QoL measures and provide a critical appraisal of their quality. There is considerable variation in how the instruments have been constructed, the categories and items included, and the methods by which their reliability and validity have been examined. In order to guide investigators in their choice of instrument, further head-to-head comparisons of measures should be undertaken. When QoL measures are compared, a Pearson or rank-order correlation coefficient is not sufficient to indicate agreement. Methods that examine agreement, rather than association, are available and should be used. In the future, we will need to demonstrate whether these measurement tools are ready for widespread implementation in clinical trials or routine patient follow-up. PMID- 9875688 TI - An evaluation of the antioxidant activity of a standardized grape seed extract, Leucoselect. AB - Coronary Artery Disease (CAD) remains the major cause of mortality and morbidity in the Western World. The oxidation of low-density lipoproteins (LDLs) by free radicals is associated with initiation of atherosclerosis and therefore, development of CAD. LDLs are protected from oxidation by antioxidants and in times of antioxidant deficiency are more likely to be oxidized. Hypercholesterolaemic patients are at a higher cardiovascular risk and may, therefore, require more antioxidant protection. Increased consumption of red wine containing antioxidants is thought to account for the lower incidence of CAD in Mediterranean countries. Red wine, although rich in antioxidants, is not suitable as routine therapy for prevention of CAD. OBJECTIVE: To evaluate the effects of a capsule formulation of an antioxidant polyphenolic extract of grapes on serum total antioxidant activity and vitamin C and E levels. METHOD: A single-blinded randomised, placebo-controlled cross-over study was undertaken in 20 young volunteers. Subjects were given two capsules containing 300 mg of grape procyanidin extracts (Leucoselect-phytosome) or placebo daily for 5 days. Blood samples were taken at the start of the study and end of the study and assayed for antioxidant activity and vitamins C and E levels. After a washout period of at least 2 weeks, the study was repeated with the second treatment. RESULTS: The extract had no effect on serum vitamins C and E levels but increased serum total antioxidant activity (TAC). On day 5, TAC increased from 408.1+/-22.9 to 453.3+/ 453.3 micromol/l trolox equivalents 1 hour postdose. CONCLUSION: The capsules increased serum antioxidant activity but the longer-term clinical implications need to be assessed in further randomised clinical trials. PMID- 9875690 TI - Influence of calcium antagonist drugs in myasthenia gravis in the elderly. AB - A number of drugs have been reported to cause neuromuscular blockade and/or to increase weakness in myasthenia gravis. We report on two patients, treated with felodipine and nifedipine for arterial hypertension, who presented with an exacerbation of their myasthenia gravis and a myasthenic syndrome or exacerbation of myasthenia gravis, respectively. The mechanism of action of calcium antagonist drugs at the neuromuscular junction is not yet well established, but it could be located at both presynaptic and postsynaptic levels. PMID- 9875691 TI - Neonatal exposure to technical methoxychlor alters pregnancy outcome in female mice. AB - This study was designed to determine the ability of female mice who were exposed neonatally to the pesticide methoxychlor (MXC) to mate, ovulate, and become pregnant upon reaching sexual maturity. One-day-old female mice (5 to 8/group) were exposed daily by intraperitoneal (ip) injection for 14 d to either sesame oil or 10 microg estradiol-17beta or 0.1, 0.5 or 1.0 mg MXC suspended in sesame oil. The MXC exposures corresponded to 14 to 71, 68 to 357, or 135 to 714 mg/kg body weight, respectively. Three months later, female mice were placed with proven breeder males and checked daily for vaginal plugs. Mated female mice were sacrificed 18 d after the appearance of a vaginal plug to evaluate pregnancy. Uteri were examined for the presence of living fetuses and/or resorption sites. Ovaries were removed and prepared for histologic evaluation and tabulation of corpora lutea. All mice from all three MXC-treated groups did in fact mate, in comparison with only one of those exposed neonatally to estradiol. Increasing the dose of MXC produced a decreased number of pregnant animals at 18 d following mating. The mean number of live fetuses/litter was reduced in the 0.5 and 1.0 mg MXC-treated groups. Corpora lutea were significantly reduced in ovaries from only the 1.0 mg MXC group and the estradiol group. No effects of treatment were seen at 0.1 mg MXC. It is concluded that neonatal exposure to MXC does not interfere with mating. Instead, significant alterations are seen in initiating and/or maintaining pregnancy. The deleterious effects on pregnancy may be due to the influence of neonatal MXC treatments on the hypothalamic-pituitary-ovarian axis as well as on possible alteration of the uterine environment. PMID- 9875692 TI - In vitro reproductive toxicity of polychlorinated biphenyl congeners 153 and 126. AB - This study was conducted to investigate the applicability of an in vitro technique for maturation, fertilization, cleavage, and growth to blastocysts of bovine oocytes to investigate reproductive toxicologic effects. During maturation, the oocytes were exposed to the di-ortho-substituted PCB congener 2,2',4,4',5,5'-CB (PCB 153) in the three concentrations 0.84 ng/mL, 8.4 ng/mL, and 84 ng/mL or to the non-ortho-substituted PCB congener 3,3'4,4',5-CB (PCB 126) in the three concentrations 1.006 pg/mL, 10.06 pg/mL, and 100.6 pg/mL and compared with control groups. PCB 153 had no effect on maturation but resulted in a reduced proportion of oocytes that cleaved at the highest concentration. There were no differences in blastocyst development among groups. PCB 126 resulted in a reduction in maturation percentage at the highest concentration and in blastocyst development at all concentrations. These results demonstrated adverse effects of PCB congeners on bovine oocytes and showed that this system can be used to evaluate toxic effects on oocytes and preimplantation-stage embryos. PMID- 9875693 TI - A longitudinal study of semen quality in pesticide spraying Danish farmers. The ASCLEPIOS Study Group. AB - It was hypothesized that occupational exposure to pesticides during a spraying season causes changes in semen quality that might be detected in a longitudinal study. We analyzed the within-person changes in semen quality and reproductive hormones across a spraying season in groups of farmers using and not using pesticides. A total of 248 men collected two semen samples (participation rate: 32%). The median sperm concentration declined significantly from the first to the second sample in both groups, but there was no statistical difference in the decline between the two groups, unadjusted or adjusted. Only minor changes were found in sperm morphology, vitality, motility, sperm chromatin denaturation (SCSA), and reproductive hormones, and the differences in changes between the two groups were nonsignificant, or, in the opposite direction to the expected. There was no relation between the changes in sperm parameters in relation to pesticide exposure variables. In conclusion, use of pesticides by Danish farmers is not a likely cause of short-term effects on semen quality and reproductive hormones. PMID- 9875694 TI - The clinical and biologic significance of serum inhibins in subfertile men. AB - Inhibin B is a marker of spermatogenesis and Sertoli cell function. The objective of this study was to evaluate the biologic significance of inhibins in subfertile men and the usefulness of inhibin B for the detection of male reproductive dysfunction. Forty-seven subfertile men were evaluated by semen analysis and clinical examination. In addition to semen analysis and hormone determinations, inhibins A and B (Serotec) in all 47 and inhibin A in 25 of these samples using another kit (Biosource) were measured. Higher inhibin B (median, range: 160.3, 81.8-328.5 pg/mL vs. 94.9, 15.6-389.7 pg/mL, P = 0.024) and lower FSH (P = 0.001) were detected in men with sperm concentrations > or =20 million/mL (n = 9), compared to oligozoospermia (sperm concentration <20 million/mL, n = 38). Inhibin B correlated significantly negatively with FSH, LH, and E2, and patient's age and positively with sperm concentration, testicular volume, and TSH. Multiple regression analysis indicated FSH, LH, E2, TSH, and age as the independent variables for inhibin B with a coefficient of determination (R) of 0.53. Simultaneous measurement of both FSH and inhibin B identified more cases with oligozoospermia than either hormone alone. Taking into account the body mass index, the age of the patient, and the indirect mixed antiglobin reaction (MAR) test result in addition to FSH and inhibin B led to the correct semen classification in 45 out of 47 cases. The simultaneous measurement of FSH and inhibin B, taking into account age, body mass index, and the indirect MAR test result appears accurate in identifying subfertility. Inhibin A is detectable in some subfertile men but its significance is not clear. PMID- 9875695 TI - Influence of teniposide (VM-26) on radiation-induced damage to mouse spermatogenesis: a flow cytometric evaluation. AB - The effect of teniposide (VM-26) 0.05 mg/kg body weight treatment on spermatogenesis of mice exposed to 0, 0.5, 1, 2, and 3 Gy gamma-radiation was evaluated flow cytometrically. Whole body irradiation with 1 to 3 Gy resulted in a significant decline in testis weight from Day 14 to 35 post-irradiation depending on the exposure dose. Treatment of mice with teniposide before irradiation advanced the decline in testicular weight by several days, especially at 3 Gy, where a significant decline in testicular weight was observed at Day 7 post-irradiation when compared with the double distilled water (DDW)+irradiation group. The relative percentage of the 2C population declined significantly in the VM-26+irradiation group in comparison with the DDW+irradiation group at various post-irradiation time periods depending on the exposure dose. A significant depletion in the relative percentage of S-phase cells was observed as early as Day 1 post-irradiation in the VM-26+irradiation group when compared with the DDW+irradiation group after exposure to 1 to 3 Gy. This decline continued up to Day 21 post-irradiation after exposure to 2 Gy. The relative percentage of primary spermatocytes showed a consistent decline after exposure to various doses of gamma-radiation in the VM-26+irradiation group when compared with the DDW+irradiation group at different time periods, with a few exceptions, especially at higher doses. The pattern of decline in the relative percentage of round spermatids was similar to that of primary spermatocytes, where a significant decline was observed at various post-irradiation time periods in the VM-26+irradiation group in comparison with the DDW+irradiation group. These changes in the relative germ cell percentages are manifested as alterations in the ratios of various germ cell populations. The 4C:2C ratio declined consistently from Day 1 to Day 70 post-irradiation in the VM-26+irradiation group at all exposure doses. Similarly, the 4C:S-phase ratio in the VM-26+irradiation group also showed a significant decline at different post-irradiation time periods when compared with the DDW+irradiation group depending on the exposure dose. The reduction observed in the relative percentages of various cell populations was higher in the combination group when compared with the DDW+irradiation controls, indicating potentiation of damage to male germ cells by teniposide treatment before irradiation. PMID- 9875696 TI - Effects of litter size on behavioral development in mice. AB - The effect of litter size on behavioral development was investigated in 1384 offspring from 114 litters of CD-1 control mice. Litters were classified in four groups of size: 5 to 7 (I), 8 to 10 (II), 11 to 13 (III), and 14 to 17 (IV). Group III was regarded as the control group. The offspring were examined for behavioral development including surface righting at Postnatal Days (PND) 4 and 7, negative geotaxis at PNDs 4 and 7, cliff avoidance at PND 7, swimming behavior at PNDs 4 and 14, and olfactory orientation at PND 14. In behavioral development, surface righting at PND 7 was significantly affected in group I. Swimming direction at PND 4 was significantly affected in group IV, and those effects showed significant tendencies to be retarded as litter size increased. Other measured parameters showed no significant effect of litter size. PMID- 9875697 TI - Comparative organophosphate-induced effects observed in adult and neonatal Sprague-Dawley rats during the conduct of multigeneration toxicity studies. AB - Five organophosphates: tribufos, oxydemeton-methyl, fenamiphos, coumaphos, and trichlorfon were evaluated for their potential to produce reproductive and neonatal toxicity following continuous dietary exposure during multigenerational reproduction toxicity studies in the Sprague-Dawley rat. Dietary concentrations were selected to demonstrate parental effects in the high dose and provide for a no-adverse effect level at the low dose. There were no clinical signs observed in the adults or neonates during either generation. Significant effects on body weight and food consumption, when observed, were typically observed only with the highest dietary concentration and were greater in the second generation. Reproductive effects, including decreased fertility and mating indices, were only observed with test compounds and at dietary concentrations demonstrating effects on body weight and/or food consumption. Similarly, pup body weight was also affected by those test compounds that produced significant maternal effects during lactation. Significant inhibition of parental cholinesterase activities (plasma, erythrocyte, and brain) was similarly observed in both generations with all test compounds, with at least the highest concentrations. In general, females demonstrated greater enzyme inhibition than the males. For example, mean PChe inhibition considering both generations and all test compounds was 74% for the females, whereas inhibition was 51% in the males. Effects on cholinesterase activities in the neonates (Lactation Day 4) were, for most test compounds, below 10% at the highest dietary concentration. However, by Lactation Day 21, inhibition of enzyme activity (considering all test compounds at the highest concentration and all enzymes) was approximately 30%. The increase in inhibition is attributed to the consumption of the treated feed during the latter stages of lactation. Considering the relative maternal (termination) and neonatal (Lactation Day 4) cholinesterase effects at the highest dietary concentration, it was observed that the effects in the neonate were, for all organophosphates tested, significantly less than those observed in the dam. PMID- 9875698 TI - Developmental toxicity assessment of arsenic acid in mice and rabbits. AB - To evaluate potential effects of exposure to inorganic arsenic throughout major organogenesis, CD-1 mice and New Zealand White rabbits were gavaged with arsenic acid dosages of 0, 7.5, 24, or 48 mg/kg/d on gestation days (GD) 6 through 15 (mice) or 0, 0.19, 0.75, or 3.0 mg/kg/d on GD 6 through 18 (rabbits) and examined at sacrifice (GD 18, mice; GD 29, rabbits) for evidence of toxicity. Two high dose mice died, and survivors at the high and intermediate doses had decreased weight gains. High-dose-group fetal weights were decreased; no significant decreases in fetal weight or increases in prenatal mortality were seen at other dosages. Similar incidences of malformations occurred in all groups of mice, including controls. At the high dose in rabbits, seven does died or became moribund, and prenatal mortality was increased; surviving does had signs of toxicity, including decreased body weight. Does given lower doses appeared unaffected. Fetal weights were unaffected by treatment, and there were no effects at other doses. These data revealed an absence of dose-related effects in both species at arsenic exposures that were not maternally toxic. In mice, 7.5 mg/kg/d was the maternal No-Observed-Adverse-Effect-Level (NOAEL); the developmental toxicity NOAEL, while less well defined, was judged to be 7.5 mg/kg/d. In rabbits, 0.75 mg/kg/d was the NOAEL for both maternal and developmental toxicity. PMID- 9875699 TI - Fetal development in alloxan-treated rats. AB - The effect of alloxan on embryo and fetal development in rats was evaluated. Alloxan was injected intraperitoneally (ip) in pregnant rats at doses of 80 to 150 mg/kg at Day 0 (day of fertilization), and 110 mg/kg at Day 4 of pregnancy. Hyperglycemia was rarely produced at alloxan doses from 80 to 100 mg/kg, and the frequency of malformations observed was low. Higher doses (110 to 150 mg/kg) caused severe hyperglycemia, and maternal or embryonic death. When 110 mg/kg was administered on Day 4 of gestation (the day before embryo implantation), all rats had resorption nodules and litters with embryos with delayed growth. We recommend the induction of diabetes mellitus on Day 4 of pregnancy for studies of diabetes gestation interaction. PMID- 9875701 TI - The sex ratio of offspring of people exposed to boron. PMID- 9875700 TI - An optimized approach for the assessment of sexual behavior in male rats. AB - The improvement and optimization of methods used to detect reproductive disorders in experimental animals are among the main challenges facing researchers in this field. The conventional method for testing male sexual behavior uses ovariectomized females rendered sexually receptive by injection of estradiol benzoate and progesterone prior to testing. The receptive females are then mated with exposed male rats during the dark phase of the cycle under dim red light followed by direct visual and momentary observation of the mating activity. The ovariectomized females may respond differently to hormonal injection (individual differences), leading to variations in the intensity of lordosis. Additionally, the data obtained by the direct visual and momentary evaluation of copulatory activity are very subjective and may produce inaccurate results. In the optimized method, the sexual cycles of female rats are determined, and only those in estrus are selected and mated with sexually experienced male rats. The mating activity is videotaped, enabling the correct observation and evaluation of the different components of mating behavior. This method fosters animal welfare by avoiding surgical intervention and enables the videotape to be kept as permanent documentation. PMID- 9875702 TI - Quantitation of 5HT2A receptor mRNA in human postmortem brain using competitive RT-PCR. AB - We describe quantitation of the absolute amounts of 5HT2A receptor mRNA in various areas of human postmortem brain using a very sensitive and specific technique: competitive reverse transcription polymerase chain reaction (RT-PCR). With this procedure, we were able to show that the highest levels of 5HT2A receptor mRNA are in cerebral cortical areas, i.e., Brodmann's areas 8, 9, 10, 18, 19, 22, 24, and 46, and that intermediate levels occur in the hippocampus, the amygdala, the nucleus accumbens, and the hypothalamus. Very low levels of 5HT2A receptor mRNA were found in the cerebellum, the thalamus, the caudate, and the putamen. Our results confirm the heterogeneous distribution of 5HT2A receptor mRNA in human postmortem brain and suggest that competitive RT-PCR is a highly sensitive and specific procedure for the quantitative measurement of 5HT2A receptor mRNA expression. The study of 5HT2A receptor mRNA in different brain areas will help clarify the regulation of 5HT2A receptors at the transcriptional level in various neuropsychiatric diseases. PMID- 9875703 TI - Identification of mammalian GFRalpha-2 splice isoforms. AB - Neurturin (NTN) belongs to a structurally related family of bioactive molecules which include glial cell-line derived neutrotrophic factor (GDNF) and perserphin (PSP). NTN exerts its effects through a multicomponent receptor system which include a receptor (GFRalpha-2) and the proto-oncogene c-RET. We report here the identification of three splice isoforms of the GFRalpha-2 receptors (GFRalpha-2a, GFRalpha-2b and GFRalpha-2c) by reverse transcription-PCR (RT-PCR). GFRalpha-2b is a novel splice variant. All three isoforms were found to be expressed in various adult murine tissues as well as in the brain of the newborn human. The identity of these isoforms were further confirmed by the isolation of the gene and the characterisation of the splice junctions. PMID- 9875704 TI - Time-dependent reversal of dentate LTP by 5 Hz stimulation. AB - Field potential recordings were made from the dentate gyrus of urethane anaesthetized rats in order to investigate the ability of 5 Hz stimulation to reverse long-term potentiation (LTP) induced by a high frequency tetanus. A 10 min train of 5 Hz was found to reverse LTP in a time-dependent fashion: as the interval between tetanus and 5 Hz was increased, LTP became progressively less susceptible to reversal. If 10 min or 30 min intervened between tetanization and 5 Hz stimulation, LTP was unaffected. These results indicate that dentate LTP in vivo exhibits a similar limited time window of vulnerability to reversal by low frequency stimulation to that previously reported in area CA1 in vitro. PMID- 9875705 TI - Muscarinic acetylcholine receptor-mediated increase of angiotensin type 2 receptor mRNA in PC12 cells. AB - The present study examined changes in the angiotensin type 2 (AT2) receptor mRNA level after carbachol exposure in PC12 cells. The AT2 receptor mRNA level increased 2- to 3-fold after 12-18 h of carbachol exposure (100 microM). The up regulation of AT2 receptor mRNA was antagonized by atropine, which is a muscarinic receptor antagonist, thus suggesting that the increase in AT2 receptor mRNA is mediated via muscarinic acetylcholine receptors. This increase is blocked by NG-nitro-L-arginine-methylester, nitric oxide (NO) synthase inhibitor, and hemoglobin NO trap. Protein kinase C (PKC) inhibitors, H-7 and calphostin C, inhibited the carbachol-induced upregulation. In addition, a G kinase inhibitor, ODQ (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one), inhibited this increase. These findings suggest that the carbachol-induced increases in AT2 receptor mRNA is regulated by the activation of NO-cGMP and/or the PKC pathway. PMID- 9875706 TI - Enhancement of slow-wave sleep by tumor necrosis factor-alpha is mediated by cyclooxygenase-2 in rats. AB - Tumor necrosis factor-alpha (TNFalpha) was infused into the subarachnoid space of the rat rostral basal forebrain, which was previously defined as a prostaglandin (PG) D2-sensitive, sleep-promoting zone. TNFalpha increased the amount of slow wave sleep (SWS), decreased that of paradoxical sleep (PS), and caused fever and anorexia. The TNFalpha-induced SWS enhancement, fever and anorexia were all blocked by co-infusion of diclofenac, a non-selective cyclooxygenase (COX) inhibitor, and by pretreatment with NS-398, a COX-2-specific inhibitor. In striking contrast, the TNFalpha-induced suppression of PS was not affected by the inhibitors. These results indicate that COX-2-mediated hyperproduction of PGs is critically involved in the enhancement of SWS, fever, and anorexia but not in the suppression of PS, caused by TNFalpha infused into the PGD2-sensitive zone. PMID- 9875707 TI - Lipopolysaccharide, central in vivo biogenic amine variations, and anhedonia. AB - Systemic administration of lipopolysaccharide (LPS), a non-specific activator of proinflammatory cytokine release from macrophages, provokes sickness characterized by anorexia, soporific effects, and disturbances of locomotor activity and exploration. In addition, endotoxin treatment may provoke an anhedonic response. Assessment of anhedonia in appetitive paradigms, however, is compromised by the anorexia provoked by the treatment. The present investigation assessed the anhedonic effects of LPS on rewarding lateral hypothalamic brain stimulation. Using a simultaneous discrimination, current titration procedure in the assessment of intracranial self-stimulation (ICSS), it was found that acute systemic administration of LPS (50 microg, 100 microg or 200 microg) reduced ICSS during the ascending sequence of current presentations, but had little effect on responding to a series of descending currents. In a parallel experiment, peripheral administration of LPS (100 microg) increased in vivo dopamine (DA) efflux from the nucleus accumbens, a region thought to be involved in goal directed responding to positively reinforcing stimuli. It is suggested that LPS alters ICSS in a manner different than that observed following stressor exposure or peripheral IL-2 treatment. Furthermore, LPS may engender an anhedonic effect (possibly secondary to sickness), and the decline of responding reflects the relation between the cost of responding given in the face of sickness and the reward received for responding. PMID- 9875708 TI - The wheat proteins puroindoline-a and alpha1-purothionin induce nodal swelling in myelinated axons. AB - The effects of two basic cysteine-rich lipid-binding proteins isolated from wheat seedlings, puroindoline-a and alpha1-purothionin, were studied on single frog myelinated axons stained with the fluorescent dye FM1-43 using confocal laser scanning microscopy. During exposure to either puroindoline-a or alpha1 purothionin (10 and 100 microM) a marked swelling of nodes of Ranvier was observed, provided NaCl was present in the external solution. It is suggested that these proteins increase the internal osmolality by forming pores in the axonal membrane and induce water influx to compensate for such an increase. Moreover, in the presence of alpha1-purothionin (100 microM), the intensity of the axonal staining with FM1-43 was increased. It is the first time, to our knowledge, that basic proteins containing domains of a cysteine-rich repeated motif are reported to produce swelling and water movements across neuronal cell membranes. PMID- 9875709 TI - Mismatch negativity: deviance detection or the maintenance of the 'standard'. AB - Electric brain responses were measured to infrequent tones that broke the frequency alternation of two tones, deviated in duration or violated both regularities (alternation and constant duration). Mismatch negativity (MMN) was elicited by both simple deviants with the duration-related MMN peaking approximately 130 ms later than the alternation-related MMN. The double deviant elicited two successive MMNs. Thus violation of each regularity elicited a separate MMN, whereas previous studies showed that multiple temporally separate deviations from a single repetitive standard elicit one MMN only. These results suggest that the primary function of the MMN-generating process is more closely related to maintaining the representation of auditory regularities than to deviance detection per se. PMID- 9875710 TI - Regeneration of the auditory pathway in adult rats by transplants of fetal brain tissue. AB - The ventral cochlear tract in the brain stem to pons was transected on one side in two groups of adult rats. In one group simple transection was performed, while in the other group tissue from embryos (E14-E16) was used to cover the lesion site. While rats receiving simple transection without transplantation showed no evidence of regeneration, in 30% of the rats receiving transplants of embryonic tissue the axons regrew beyond the transected site and regenerated into the denervated side and terminated at the normal targets. The present findings contradict the widely held view that the adult mammalian central auditory system cannot be restored following damage. PMID- 9875711 TI - Failure of dominant left-hemispheric activation to right-ear stimulation in schizophrenia. AB - Schizophrenia is associated with an absence of the lateralizations that typify the human brain. Previous evidence emphasized structural changes, particularly reduced asymmetry in extension and surface of the planum temporale, although gross structural deviations occur only in a minority of patients. The present study describes an absence of lateralization on a robust functional measure that characterized schizophrenia patients: healthy subjects but not schizophrenics displayed a contralateral left-hemispheric dominance of the auditory evoked magnetic field to right-ear auditory stimulation. Absence of contralateral dominance in response to auditory stimuli among schizophrenia patients may indicate a failure to establish unequivocal left-hemispheric dominance of the phonological loop as hypothesized by Crow. PMID- 9875712 TI - Retroviral-mediated transfer of the galactocerebrosidase gene in neural progenitor cells. AB - Globoid cell leukodystrophy (GCL or Krabbe disease) is a recessive disease caused by mutations of the lysosomal enzyme galactocerebrosidase (GALC) and twitcher is the murine model of GCL. We have prepared retroviral packaging cell lines to transduce the GALC gene. Retroviral transduction restored GALC activity in GCL fibroblasts and increased such activity to very high levels in immortalized neural progenitor cells (ST14A cells). GALC activity was also normalized in twitcher fibroblasts co-cultured with ST14A cells over-expressing GALC, demonstrating that this enzyme is secreted and can be imported efficiently by GALC-deficient cells. These results give the necessary background to evaluate the therapeutic effect in twitcher of brain grafting of neural progenitor cells engineered to release high levels of GALC. PMID- 9875713 TI - Striatal dopamine-glutamate interactions reflected in substantia nigra reticulata firing. AB - To gain insight into the role of striatal dopamine in basal ganglia functioning, dopaminergic drugs alone and in combination with the glutamate receptor agonist kainic acid were infused in the lateral striatum via a microdialysis probe, while single-unit recordings of substantia nigra reticulata neurons were made in chloral hydrate-anaesthetized rats. Striatal infusion of dopaminergic drugs did not significantly affect the firing rate of substantia nigra reticulata neurons, which was related to the low activity of striatal cells under basal conditions, illustrated by the lack of effect of striatal infusion of TTX on substantia nigra reticulata activity. Under glutamate-stimulated conditions, striatal infusion of d-amphetamine potentiated the inhibition of substantia nigra reticulata neurons induced by striatal kainic acid. Thus, under stimulated but not basal conditions, the modulatory role of dopamine in the striatum could be demonstrated. Dopamine potentiated the inhibitory effect of striatal kainic acid on the firing rate of the basal ganglia output neurons. PMID- 9875714 TI - The effect of extracellular matrix on the growth of mouse olfactory tissue in vitro. AB - The aim of this study was to compare the capacity of extracellular matrix (ECM) components to support the growth and differentiation of olfactory tissue in vitro. Reconstituted basement membrane matrix (RBM) supported profuse neurite outgrowth from explants of olfactory bulb (OB), with no non-neuronal cell migration. In contrast, nasal tissue (NT) explants cultured on RBM produced profuse fibroblastic outgrowth, but no neurite outgrowth from olfactory sensory neurones (OSN). Three-dimensional collagen gels, on the other hand, supported extensive outgrowth of both non-neuronal and neurite outgrowth from OB and NT explants, mature OSNs in the latter being identified using antibodies against olfactory marker protein (OMP). These culture systems represent useful tools for investigating potential tropic influences of target tissue. PMID- 9875715 TI - Three-dimensional modeling of static vestibulo-ocular brain stem syndromes. AB - Static vestibulo-ocular brain stem syndromes characterized by skew deviation, a vertical disconjugacy of the eyes, and ocular torsion are the result of a vestibular tone imbalance in the frontal (roll) plane. Similar physiological changes in static eye position, ocular counter-roll and conjugated deviations of vertical eye position, are caused by the influence of gravity mediated by the utricles. These observations prompted our approach with the model described here: based on the known deviations of static eye position, we devised a three dimensional mathematical model of otolith-ocular function including detailed brain stem anatomy. This model is able to explain and predict the differential effects of unilateral and bilateral peripheral or central vestibular lesions on static eye position in roll, pitch, and yaw planes. PMID- 9875716 TI - Calretinin and calbindin D28K immunoreactivity in the superficial layers of the rabbit superior colliculus. AB - Calretinin and calbindin D28K were localized in the superficial layers of rabbit superior colliculus (SC). Calretinin and calbindin D28K-immunoreactive (-IR) neurons were concentrated in the upper superficial gray layer. Calretinin-IR fibers were found in the optic layer. The majority of calretinin-IR cells were small- to medium-sized vertical fusiform neurons and neurons with round or stellate-shaped somas with small varicose dendrites. The morphology of calbindin D28 K-IR neurons was different from that of calretinin neurons. Anti-calbindin D28K-IR neurons usually had fusiform cell bodies and a thick primary dendrite with small branches forming a dendritic bouquet. Two-color immunofluorescence revealed that no cells expressed both proteins. Following unilateral enucleation a marked reduction of calretinin-IR fibers in the contralateral side to the enucleation was found. Enucleation appeared to have no effect on the cell bodies labeled with either protein. The results suggest the anti-calretinin immunoreactivity in the superficial layer of rabbit SC contrasts starkly with that of other animals. PMID- 9875717 TI - 'Musical brain' revealed by high-field (3 Tesla) functional MRI. AB - The cortical areas subserving music literacy were investigated using high-field (3 Tesla) functional magnetic resonance imaging (fMRI). The activation pattern associated specifically with music score reading was compared with that associated with reading text in a subject's primary and secondary language. While the areas of activation were predominantly identical for all three reading modalities, there were areas within the occipital cortex activated exclusively by music score reading. Grand analysis of the activation patterns of eight pianists unequivocally identified that the principal cortical area needed for music literacy is the cortex flanking the right transverse occipital sulcus (musical brain). PMID- 9875718 TI - Mitotic index and Alzheimer's disease. AB - Alzheimer's disease (AD), a progressive neurodegenerative disorder, is diagnosed definitively by increased numbers of beta-amyloid plaques and neurofibrillary tangles in brain biopsy or autopsy specimens. There are no simple straightforward laboratory tests currently available for clinical diagnosis. We have found consistent reduction in mitotic index levels in skin fibroblast cultures from AD individuals compared with age- and sex-matched controls. These differences were enhanced by overnight exposure to colcemid (p = 0.04). Results suggest that mitotic index in skin fibroblasts cultures should be further investigated as a potential diagnostic indicator for AD. PMID- 9875719 TI - Minimal effect of brain temperature changes on glutamate release in rat following severe global brain ischemia: a dialysis electrode study. AB - Using a dialysis electrode, we recently developed an oxygen-independent system for real-time measurement of the glutamate concentration in the extracellular space ([Glu]e) during ischemia. This system allows separate evaluation of intra ischemic biphase [Glu]e elevation, i.e. release from synaptic vesicles (1st phase), reversed uptake of glutamate from metabolic pools in neuronal cells (2nd phase), and post-ischemic glutamate re-uptake in ischemia-reperfusion models. Using the system, we attempted to clarify the relationship between biphase glutamate release and brain temperature in a model of acute global ischemia produced by transecting both carotid arteries. Our results showed that, in contrast to mild hyperthermia, hypothermia did not inhibit the 1st phase of [Glu]e release, and changes in intra-ischemic brain temperature had a minimal effect on the 2nd phase of [Glu]e elevation during severe acute ischemia. These findings, together with our previous data, indicate that brain temperature change in the intra-ischemic period plays an important role in disturbance of the glutamate re-uptake system during ischemia. PMID- 9875720 TI - COX-2 inhibitor prevents the development of hyperalgesia induced by intrathecal NMDA or AMPA. AB - Cyclooxygenase (COX)-2 is constitutively expressed in the superficial dorsal horn of the spinal cord, but its role in the spinal cord is still unclear. We examined the effect of intrathecally administered NS398, a selective COX-2 inhibitor and indomethacin, an non-selective COX-1 and COX-2 inhibitor, on the development of thermal hyperalgesia induced by the activation of NMDA or AMPA receptors. Intrathecal injection of either NS398 or indomethacin equally blocked the development of thermal hyperalgesia induced by intrathecal injection of either NMDA or AMPA in a dose-dependent manner. These data suggest that COX-2 plays an important role in spinal thermal nociceptive transmission when neurons in the spinal cord are facilitated by NMDA or AMPA. PMID- 9875721 TI - C-terminal fragment of amyloid precursor protein inhibits calcium uptake into rat brain microsomes by Mg2+-Ca2+ ATPase. AB - Numerous lines of evidence suggest that some of the neurotoxicity associated with Alzheimer's disease (AD) is due to proteolytic fragments of the amyloid precursor protein (APP). Most research has focused on the amyloid beta peptide (A beta). However, the possible role of other cleaved products of APP is less clear. In this study, the effects of a recombinant carboxy terminal 105 amino acid (CT105) fragment of APP on the calcium uptake by endoplasmic reticulum Mg2+-Ca2+ ATPase, the major mechanism for sequestering calcium in this organelle, were investigated. We found that CT 105 is a potent inhibitor of Mg2+-Ca2+ ATPase of endoplasmic reticulum, whereas A beta shows no effect. These results demonstrate that CT 105 inhibits the ability of brain microsomes to sequester calcium and suggest that this inhibitory effect of CT 105 may contribute to disruption of intracellular calcium concentration, possibly being involved in inducing the neural toxicity characteristic of AD. PMID- 9875722 TI - Centrally applied NPY mimics immunoactivation induced by non-analgesic doses of met-enkephalin. AB - Neuropeptide Y (NPY) and endogenous opioids (EOPs) such as methionine-enkephalin (Met-enk) regulate similar physiological responses, but it is not known whether nociceptive and immune responses also show analogy after intracerebroventricular (i.c.v.) application. Dose-response studies show that Met-enk stimulates the blood granulocyte and splenic natural killer (NK) cell function of Lewis rats at a low dose (10(2) ng/kg, i.c.v.), whereas a high dose (10(5) ng/kg) causes suppression of innate immune functions associated with analgesia in the hot-plate test. At 15 min, 1 h and 24 h after i.c.v. application, both Met-enk (10(2) ng/kg) and NPY (1 ng/kg) produced similar effects: An initial suppression of innate immune function was followed by a long lasting stimulatory action on cell functions and serum interleukin-6 (sIL-6) levels. Thus, central NPY application resembles Met-enk-induced immunostimulation at doses not affecting nociception, suggesting an involvement of both peptides in shaping stress-induced immunomodulation of the non-analgetic form, possibly via activation of a common immunomodulatory effector mechanism. PMID- 9875723 TI - A potassium current in guinea-pig outer hair cells activated by ion channel blocker DCDPC. AB - Fenamate compounds have been reported to inhibit ion channels in a number of tissues, including a non-selective cation channel in the mammalian outer hair cell (OHC). We have further investigated the effects of 3'-5 dichlorodiphenylamine-2-carboxylic acid (DCDPC) on OHC currents using the whole cell configuration of the patch clamp technique. Extracellular application of 10 microM DCDPC rapidly and reversibly activated an inward current at hyperpolarized potentials. The DCDPC-activated current appeared in the shorter OHCs from the basal turns of the cochlea. The reversal potential of the inward current was dependent on the external K+ ion concentration. An outwardly rectifying K+ current, found predominantly in OHCs from apical turns, was reversibly inhibited by DCDPC. These results suggest that DCDPC has a significant effect on OHC physiology at all tonotopic locations along the basilar membrane and so may have implications for cochlear function during fenamate intake. PMID- 9875724 TI - Are aminoglycoside antibiotics excitotoxic? AB - Guinea pigs received gentamicin to induce a profound hearing loss (61 dB auditory threshold shift at 18 kHz). Concomitant administration of maleic or tartaric acid dissolved in dimethyl sulfoxide (DMSO) significantly reduced the threshold shift to < 40 dB. The results have several important implications. First, they support the hypothesis of a free-radical mechanism of gentamicin toxicity since the protective compounds are metal chelators and scavengers. Second, they caution against these and similar chemicals, commonly found in drug preparations, as vehicles in tests of aminoglycoside toxicity. For example, a recent study by others describing attenuation of aminoglycoside ototoxicity by NMDA antagonists may have been influenced by the presence of maleate, tartrate and DMSO. Third, they suggest simple antioxidants as a potentially efficient and inexpensive clinical prophylaxis of aminoglycoside-induced hearing loss. PMID- 9875725 TI - Psilocybin induces schizophrenia-like psychosis in humans via a serotonin-2 agonist action. AB - Psilocybin, an indoleamine hallucinogen, produces a psychosis-like syndrome in humans that resembles first episodes of schizophrenia. In healthy human volunteers, the psychotomimetic effects of psilocybin were blocked dose dependently by the serotonin-2A antagonist ketanserin or the atypical antipsychotic risperidone, but were increased by the dopamine antagonist and typical antipsychotic haloperidol. These data are consistent with animal studies and provide the first evidence in humans that psilocybin-induced psychosis is due to serotonin-2A receptor activation, independently of dopamine stimulation. Thus, serotonin-2A overactivity may be involved in the pathophysiology of schizophrenia and serotonin-2A antagonism may contribute to therapeutic effects of antipsychotics. PMID- 9875726 TI - Endothelin in perivascular nerves. An electron-immunocytochemical study of rat basilar artery. AB - The endothelium is recognized as a major source of endothelin in the vasculature. In this report we show for the first time that endothelin can be demonstrated with immunoelectronmicroscopy in perivascular nerves. About 36% of the axon profiles (varicosity/intervaricosity) examined in the rat basilar artery showed immunolabelling with a polyclonal antibody to endothelin-1. Endothelin-labelled axon varicosities were characterized by the presence of small spherical agranular vesicles (42 +/- 1 nm); some varicosities also contained large granular vesicles (92 +/- 5 nm) with labelled cores. Immunolabelling was mostly distributed in the axoplasm and in association with the membrane of synaptic vesicles (the lumen of the small agranular vesicles was immuno-negative). The presence of endothelin in perivascular nerves of the basilar artery suggests a neuronal as well as endothelial role in the physiological control of the vessel wall. PMID- 9875727 TI - Cerebellar dysfunctions of temporal processing in the seconds range in humans. AB - The roles of the basal ganglia and cerebellum in timing remain subject to debate. It has been suggested that temporal range may dissociate them, since cerebellar research has focused on intervals of < 1 s, compared with many seconds used in much basal ganglia research. Here we show increased but scalar variability of time estimates in patients with focal lesions of the lateral cerebellar cortex and nuclei when trained to remember durations in the seconds range, compared with patients with lesions of the mesial cerebellum and vermis. This distortion is discussed on the basis of previously reported cerebellar deficits in different time ranges and contrasted to distortions occurring in patients with basal ganglia lesions performing similar tasks over similar time ranges. PMID- 9875728 TI - Enhanced AP-1 binding in brain induced by D1 and D2 agonists in methamphetamine sensitized rats. AB - The activator protein-1 (AP-1) binding activities induced by a separate challenge with SKF38393 and quinpirole after 1 weeks' abstinence from chronic methamphetamine (4 mg/kg/day, 14 days) were increased significantly in the striatum, nucleus accumbens and cingulate cortex compared with the saline-treated controls. Quinpirole-, but not SKF38393-induced AP-1 binding activities were still significantly higher after a 4-week abstinence period in the chronic methamphetamine group than in the chronic saline control group. Downward sniffing, which occurred following a quinpirole-challenge, was significantly intensified after both a 1 and 4 weeks' abstinence from chronic methamphetamine. These results indicate that chronic administration of methamphetamine induces alterations of the interactions of dopamine D1 and D2 receptors which are reflected as enhanced AP-1 binding activities. PMID- 9875729 TI - MK-801 and male odours induce c-fos expression in the AOB of juvenile female mice. AB - The odours of adult males, which accelerate the timing of puberty of female mice, activate c-fos in the accessory olfactory bulb (AOB). To test the hypothesis that NMDA receptors are involved in the male odour-induced increase in c-fos expression, we studied the effects of the non-competitive NMDA receptor agonist MK-801 on male odour-induced c-fos expression in the AOB of juvenile female mice. Surprisingly, MK-801 increased FOS-like immunoreactivity (FLI) within the AOB in the absence of male odour and had no effect on male odour-induced c-fos expression. We suggest that MK-801 increases AOB mitral cell activity by disinhibiting GABAergic granule cells, resulting in increased c-fos expression throughout the AOB. PMID- 9875730 TI - Mutation screening in exons 3 and 4 of alpha-synuclein in sporadic Parkinson's and sporadic and familial dementia with Lewy bodies cases. AB - Recently it has been reported that a missense G(88)C mutation within exon 3 and a missense G(209)A mutation within exon 4 of the alpha-synuclein gene were linked to familial Parkinson's Disease (PD). We decided to investigate if these and any other mutations in exons 3 and 4 of the alpha-synuclein gene could be detected in sixty two sporadic PD and dementia with Lewy bodies (DLB) patients. Four cases of familial DLB were also studied, two of which were from the same family. Single stranded conformational polymorphism, DNA sequencing analyses and PCR-RFLP of exons 3 and 4 failed to reveal any nucleotide changes. However, three nucleotide differences occurred in the intron 4 sequence compared to the published sequence. This study adds further support to the idea that these particular mutation in the alpha-synuclein gene are a rare case of PD and now, as we have shown here, also of DLB. PMID- 9875731 TI - Parieto-occipital approximately 10 Hz activity reflects anticipatory state of visual attention mechanisms. AB - High-density eeg recordings revealed sensory specific modulation of anticipatory parieto-occipital approximately 10 Hz oscillatory activity when visually presented word cues instructed subjects in an intermodal selective attention paradigm. Cueing attention to the auditory features of imminent compound audio visual stimuli resulted in significantly higher approximately 10 Hz amplitude in the period preceding onset of this stimulus than when attention was cued to the visual features. We propose that this parieto-occipital approximately 10 Hz activity reflects a disengaged visual attentional system in preparation for anticipated auditory input that is attentionally more relevant. Conversely, lower approximately 10 Hz activity during the attend-visual condition may reflect active engagement of parieto-occipital areas in the anticipatory period. These results support models implicating parieto-occipital areas in the directing and maintenance of visual attention. PMID- 9875732 TI - Identification of two novel diurnal genes by screening of a rat brain cDNA library. AB - While the hypothalamus is fundamental for sleep and circadian regulation, the molecular mechanisms involved are poorly understood. We have used a differential gene expression technique to identify hypothalamic genes which have altered expression in rat sleep periods. Complex cDNA probes from rat hypothalami removed at Zeitgeber times 4 and 15 were hybridised to rat brain cDNA library girds. From 30 differentially expressed clones, six were further analysed and two were confirmed to exhibit increased expression at Zeitgeber time 4. A Northern blot hybridization of brain, heart, kidney, lung, testis and skin mRNA showed that both clones were brain specific. Therefore, we have identified two novel brain specific diurnally expressed hypothalamic genes. Both genes may have roles in sleep or circadian regulation. PMID- 9875733 TI - Reward-quality dependent anticipation in rat nucleus accumbens. AB - Single unit recording in rat nucleus accumbens (NAcc) was used to ascertain NAcc neuronal activity in mediating of reward including its anticipation. Of the 103 neurons investigated, 63% showed some response in connection with the task activity. Of these, 20 units responded during delivery of the primary reward (food and/or water) and five responded during the time period preceding reward if the reward was delayed (four to food, one to water). These result suggest that NAcc neurons responded not only to the delivery of primary reward and task inducing anticipation of reward, but also represent the difference of reward quality between food and water specifically. PMID- 9875734 TI - Serum deprivation-induced apoptosis in cultured hippocampi is prevented by kainate. AB - Serum deprivation of hippocampal organotypic cultures induced cell death within 6 h in dentate gyrus granule cells and hilar interneurons whereas neurons from other hippocampal regions were spared. Dying neurons exhibited condensed chromatin in the nuclei, as revealed by cresyl violet, Hoescht staining, and electron microscopy. Cell death was abolished by cycloheximide. KA, an agonist of AMPA/KA receptors that induces depolarization, also prevented neuronal death. This effect was antagonized by the AMPA/KA receptor antagonist DNQX, but not by APV, an antagonist of NMDA receptors. PTX, a GABA(A) receptor antagonist, reduced neuronal death by 50% after serum withdrawal. These data indicate that protein synthesis-dependent programmed cell death (PCD) occurs in the dentate gyrus upon trophic support withdrawal and suggest that neuronal activity contributes to cellular homeostasis. PMID- 9875735 TI - Blockade of adenosine A2A receptors by SCH 58261 results in neuroprotective effects in cerebral ischaemia in rats. AB - Blockade of adenosine receptors can reduce cerebral infarct size in the model of global ischaemia. Using the potent and selective A2A adenosine receptor antagonist, SCH 58261, we assessed whether A2A receptors are involved in the neuronal damage following focal cerebral ischaemia as induced by occluding the left middle cerebral artery. SCH 58261 (0.01 mg/kg either i.p. or i.v.) administered to normotensive rats 10 min after ischaemia markedly reduced cortical infarct volume as measured 24 h later (30% vs controls, p < 0.05). Similar effects were observed when SCH 58261 (0.01 mg/kg, i.p.) was administered to hypertensive rats (28% infarct volume reduction vs controls, p < 0.05). Neuroprotective properties of SCH 58261 administered after ischaemia indicate that blockade of A2A adenosine receptors is a potentially useful biological target for the reduction of brain injury. PMID- 9875736 TI - Random dots blinking: a new approach to elucidate the activities of the extrastriate cortex in humans. AB - To investigate cortical activities related to the visual recognition of characters, we recorded the magnetoencephalography (MEG) in six normal subjects who were encouraged to discriminate capital English letters displayed for a brief period. To reduce the primary responses evoked by the luminance change in the striate cortex (V1), we used a novel stimulus method, random dots blinking (RDB), by means of the temporal changes of patterns using a large number of small random dots. Along with the MEG recording, we also measured the discrimination accuracy rate (%) to know how well the subjects recognized the letters. One clear component, about 300 ms in peak latency, was identified in all six subjects. Its peak amplitude and the discrimination accuracy rate increased similarly as the character display duration became longer. Its signal source was estimated in the extrastriate cortex, around the fusiform gyrus, in the right hemisphere. We suspect that the activity in these cortical areas has strong relation to the conscious perception of characters. PMID- 9875737 TI - Novel insertion in the KSP region of the neurofilament heavy gene in amyotrophic lateral sclerosis (ALS). AB - The abnormal assembly and accumulation of neurofilaments (NF) in the perikarya and proximal axons of motor neurones is a characteristic of ALS. Deletions in the KSP repeat region of the NF-H gene have previously been reported in seven patients with sporadic ALS. Here we report the identification of a novel 84 bp insertion in the NF-H gene. This leads to an extra four KSP repeat elements in a highly conserved repetitive region of the gene. Although neurofilament mutations are only associated with a very small proportion of ALS cases, this insertion provides further support of a role for neurofilaments in the pathogenesis of ALS. PMID- 9875738 TI - Lithium treatment causes gliosis and modifies the morphology of hippocampal astrocytes in rats. AB - The biological basis of the clinical efficacy of lithium in the treatment of mental illness has been extensively studied in neurones, but little is known about the effects of the drug on glia. Recently we showed that treatment of rats with clinically relevant doses of lithium chloride results in a 35% increase in the immunocontent of the astrocyte marker GFAP in the hippocampus. Here we studied the cytology of this phenomenon. Rats were treated for 4 weeks with a lithium diet which resulted in serum Li+ concentrations of 0.6-1.2 mmol/l. GFAP immunocytochemistry of the hippocampus revealed a mild gliosis in the CA1 area and the dentate gyrus which was associated with a change in the orientation of astrocytic processes. In control animals astrocyte processes were mainly orientated perpendicular to the stratum pyramidale, whereas in treated animals the cells were predominantly stellar in appearance. PMID- 9875739 TI - Cloning and expression pattern of a murine semaphorin homologous to H-sema IV. AB - The semaphorin/collapsin family of proteins comprises molecules thought to be important for the guidance of growing axons. All members of this family, which includes both secreted and cell-surface molecules, share a conserved domain of approximately 500 amino acids. Here, we report the cloning of a novel murine semaphorin, termed M-sema IV. It displays 96.3% identity to the human semaphorin H-sema IV and is therefore likely to be the respective murine homologue. In addition, an isoform was identified, which contains an additional 31 amino acids in the semaphorin domain. M-sema IV appears to be expressed ubiquitously in adulthood. During embryogenesis, in situ hybridization revealed M-sema IV expression in subregions of the central nervous system and various other tissues like skin, kidney, lung and intestine. PMID- 9875740 TI - Short-latency ocular following of motion by monkeys during a fixation task. AB - Short-latency ocular following responses in monkeys, induced by first- and second order motion during a fixation task, were sensitive to stimulus size, retinal eccentricity and motion duration. Latency of eye movements was no different for first- or second-order motion and simply marked the occurrence of movement in the visual field. Second-order motion produced higher velocity eye movements than did first-order motion. The velocity of eye movements was influenced by the orientation of second-order pattern components as well as their conspicuous energy. Even when partially suppressed by a fixation task, involuntary eye movements during the initial pursuit remain sensitive to several properties of visual motion. PMID- 9875741 TI - Central nervous system action of melatonin on gastric acid and pepsin secretion in pylorus-ligated rats. AB - We recently demonstrated that centrally administered melatonin at low doses inhibits the induction of gastric lesions by water-immersion restraint stress. To investigate the mechanism of the potent anti-ulcer action of melatonin, the central nervous system (CNS) effects of melatonin on gastric acid and pepsin secretion were studied in conscious pylorus-ligated rats. Intracisternal (i.c.) melatonin (1-100 ng) dose-dependently decreased acid and pepsin output, while a higher i.p. dose (1 microg) had no inhibitory effect. The i.c. melatonin did not change serum gastrin concentrations. Serum melatonin concentrations at 1 and 4 h after i.c. administration of 10-100 ng melatonin did not differ from those in rats receiving i.c. vehicle. The present results suggest that melatonin administered centrally modulates the secretion of gastric acid and pepsin which may explain, at least in part, the protective, anti-stress role of melatonin in the gastric mucosa observed in our previous study. PMID- 9875742 TI - Circulatory support with pneumatic paracorporeal ventricular assist device in infants and children. AB - BACKGROUND: Mechanical circulatory support in intractable heart failure in children has been limited to centrifugal pumps and extracorporeal membrane oxygenation: Since 1990 small adult-size pulsatile air-driven ventricular assist devices "Berlin Heart" (VAD) and, since 1992 miniaturized pediatric VAD (12, 15, 25, 30 mL pumps), have been used in our institution. Since 1994 the blood contacting surfaces of the device system have been heparin-coated. In this report the experiences with VAD support in 28 children are presented. METHODS: In 28 children-ages between 6 days and 16 years-the Berlin Heart VAD has been applied for periods of between 12 hours and 98 days (mean, 16.9 days) aiming at keeping the patient alive and allowing for recovery from shock sequelae until later transplantation or myocardial recovery. There were three groups. Group I: with primary intention of "bridge-to-transplantation" in various forms of cardiomyopathy (n = 13) or chronic stages of congenital heart disease (n = 5). Group II: "Rescue" in intractable heart failure early after corrective surgery for congenital heart disease (n = 4) or in early graft failure after a heart transplantation (n = 1). Group III: "Acute myocarditis" (n = 5) aiming at either myocardial recovery or transplantation. Twelve were brought to the operating room under cardiac massage and 25 had been on the respirator for more than 24 hours. RESULTS: Twelve patients died on the system from sequelae of profound shock multiorgan failure, sepsis, loss of peripheral circulatory resistance-or from hemorrhagic complications (n = 4) or brain death (n = 1). Thirteen patients (groups I and III) were transplanted after support periods of between 3 and 98 days with 7 long-term survivors living now up to 7.5 years (mean, 4.4 years). Three patients (groups II and III) were weaned from the system with two long-term survivors (both in group III). There were no patients in group II who survived and the "rescue" indication has been discarded for VAD since 1992. Such patients are since treated by extracorporeal membrane oxygenation (ECMO) in our institution. Out of the 8 patients placed on VAD during 1996 and 1997, 7 were successfully supported until transplantation or weaning. Thirteen patients were extubated and mobilized on the system. Whereas with the earlier systems thrombi in the blood pumps were seen in 15 instances and 2 patients suffered from thromboembolic complications, no thrombotic events occurred with the heparin coated systems. CONCLUSIONS: After accumulating clinical experience and several technical improvements since 1990 the use of the pediatric Berlin Heart VAD has matured into a reliable and safe system to keep patients with otherwise intractable heart failure alive until complete myocardial recovery is reached or transplantation becomes feasible. Whereas heart failure early after cardiac operation is now primarily treated by ECMO, acute myocarditis appears to be a promising precondition for complete cardiac recovery during VAD support. PMID- 9875743 TI - Strategies of myocardial protection for operation in chronic model of cyanotic heart disease. AB - BACKGROUND: Cyanotic congenital hearts have an increased susceptibility to ischemia and subsequent reperfusion. The role of platelet-activating factor antagonism and mechanical neutrophil depletion with leukocyte-depleting filters for control of ischemia-reperfusion injury was assessed in corrective surgical procedures for cyanotic heart disease. METHODS: A swine model of cyanotic heart disease was evaluated with three study groups: a control group; a group given a platelet-activating factor antagonist (PAFA group); and a group with leukocyte depleting filtration (LDF group). The cyanotic model was created with a left atrial appendage-pulmonary artery fistula with peripheral banding through a left anterior thoracotomy in weanling swine. The experimental procedure was performed 5 to 7 weeks later when body weight was greater than 20 kg and oxygen saturation was 85% or less. The corrective procedure was performed through a median sternotomy on cardiopulmonary bypass with repair of the shunt. Myocardial protection was accomplished with hypothermic blood-crystalloid (4:1) cardioplegia; the period of ischemic arrest was 90 minutes. In the PAFA group, the platelet-activating factor antagonist CV-6209 was delivered intravenously 15 to 20 minutes before aortic cross-clamping. In the LDF group, Pall leukocyte depleting filters were used in the CPB arterial line. Hemodynamic data were taken before operation and 10 and 30 minutes after CPB with impedance ventriculography. RESULTS: There were four deaths in the control group within 30 minutes after CPB; all animals in the treated groups survived longer than 60 minutes (p < 0.05). The ventricular assessment of end-systolic elastance revealed superior performance in the LDF group 30 minutes after CPB compared with the control group (p < 0.05) (controls, 4.0+/-9; PAFA group, 6.5+/-3.7; and LDF group, 12.0+/-4.6). CONCLUSIONS: Both leukocyte-depleting filters and platelet-activating factor antagonism provided myocardial protection, and the filters afforded superior postoperative myocardial contractility. PMID- 9875744 TI - Relentless pulmonary vein stenosis after repair of total anomalous pulmonary venous drainage. AB - BACKGROUND: Progressive stenosis of the pulmonary veins after repair of total anomalous pulmonary venous drainage is frequently refractory to surgical therapy. METHODS: Retrospective review of 170 consecutive patients treated for total anomalous pulmonary venous drainage identified 13 patients with postrepair pulmonary vein stenosis. Preoperative and operative data were analyzed to define the patterns of progression and efficacy of surgical techniques. RESULTS: Seventeen reoperations were performed in 13 patients. Postrepair pulmonary vein stenosis was most common in the infracardiac and mixed subtypes (p < 0.02). All 4 patients with unilateral stenosis, 2 of whom had progression of stenosis resulting in nearly complete unilateral pulmonary vein occlusion, survived. Six of 9 patients with bilateral disease died (p < 0.05 versus unilateral); 2 of the 3 survivors were repaired with a novel technique creating a sutureless neoatrium without evidence of restenosis at 1.8 years' follow-up. Stenting was uniformly unsuccessful. CONCLUSIONS: In unilateral stenosis, progression of disease may be survivable with loss of single-lung perfusion. Although bilateral disease is lethal in most cases, creation of a sutureless neoatrium has demonstrated short term freedom from disease progression. PMID- 9875745 TI - Surgical management of total anomalous pulmonary venous drainage: impact of coexisting cardiac anomalies. AB - BACKGROUND: Recent reports have cited improving results for surgical management of isolated total anomalous pulmonary venous drainage. Complex cases (with other cardiac anomalies) are less frequently reported and are associated with higher mortality. METHODS: Retrospective review identified 170 consecutive patients treated for total anomalous pulmonary venous drainage from 1982 to 1996: 44 cases were "complex" (with significant associated cardiac lesions) and 126 cases were "simple." RESULTS: Operative mortality for simple cases decreased from 26% to 8%, and mortality for complex cases remained constant at 52%. Age, size, and the presence of atrial isomerism were univariate predictors of mortality. Multivariable analysis identified only univentricular hearts and associated cardiac lesions as predictors of operative mortality. Pulmonary artery (n = 16) and arteriopulmonary (n = 7) shunting strategies for complex cases resulted in less than 30% long-term survival. CONCLUSIONS: Despite improvement in survival for simple cases, management of total anomalous pulmonary venous drainage with single-ventricle hearts or other associated cardiac lesions remains problematic. PMID- 9875746 TI - Neonatal piglet model of intraaortic balloon pumping: improved efficacy using echocardiographic timing. AB - BACKGROUND: Pediatric intraaortic balloon pumping (IABP) has met with little success because of technical difficulty in tracking rapid heart rates. This study was designed to evaluate the efficacy of M-mode echocardiography for IABP timing in a neonatal piglet model. METHODS: Two groups of piglets underwent mitral valve avulsion to create a model of shock. Group 1 (n = 8; mean weight, 7.7+/-1.8 kg) underwent IABP timed with both the ascending aortic pressure and M-mode echocardiogram. Group 2 (n = 6; mean weight, 7.5+/-1.4 kg) underwent two separate periods of IABP: one with echocardiographic timing and the second using standard timing points from the femoral arterial pressure tracing and electrocardiogram. Measurements included ascending aortic flow, left anterior descending arterial flow, ascending aortic pressure, left atrial pressure, and heart rate. RESULTS: Mitral valve avulsion produced a shock model with a significant decrease in mean aortic pressure and aortic flow and a significant increase in left atrial pressure and heart rate. Compared with the shock state, IABP in group 1 animals resulted in a significant increase in aortic flow (353+/-152 versus 454+/-109 mL/min; p < 0.05) and a significant decrease in left atrial pressure (23+/-6 versus 17+/-7 mm Hg; p < 0.05). Group 2 animals with echocardiogram-timed IABP had significantly increased aortic flow (365+/-106 versus 458+/-107 mL/min; p < 0.05) and mean aortic pressure (43+/-11 versus 52+/-8 mm Hg; p < 0.05). However, standard-timed IABP failed to show any improvement. CONCLUSIONS: In piglets with rapid heart rates, echocardiogram-timed IABP results in increased aortic flow and pressure and decreased left atrial pressure compared with standard-timed IABP. PMID- 9875747 TI - Double patch closure of ventricular septal defect with increased pulmonary vascular resistance. AB - BACKGROUND: Closure of a large ventricular septal defect (VSD) in children with elevated pulmonary vascular resistance is associated with significant morbidity and mortality. Pulmonary hypertensive episodes continue to be a major cause of postoperative morbidity and mortality. We designed a fenestrated flap valve double VSD patch in an effort to decrease the morbidity and mortality associated with the closure of a large VSD with elevated pulmonary vascular resistance. METHODS: Eighteen children (mean age, 5.7 years) with a large VSD and elevated pulmonary vascular resistance (mean, 11.4 Wood units) underwent double patch VSD closure using moderately hypothermic cardiopulmonary bypass and cardioplegic arrest. The routine VSD patch was fenestrated (4 to 6 mm) and on the left ventricular side of the patch, a second, smaller patch was attached to the fenestration along its superior margin before closure of the VSD. RESULTS: All children survived operation and were weaned from inotropic and ventilator support within 48 hours postoperatively. Postoperative pulmonary artery pressures were significantly lower than preoperative values. One child died 9 months postoperatively. CONCLUSIONS: Closure of a large VSD in children with elevated pulmonary vascular resistance can be performed with low morbidity and mortality when a flap valve double VSD patch is used. PMID- 9875748 TI - Late results of bioprosthetic tricuspid valve replacement in Ebstein's anomaly. AB - BACKGROUND: Historically, porcine bioprosthetic valves have poor durability in pediatric patients; nearly half will require replacement within 5 years. However, our early experience with patients having Ebstein's anomaly suggests that tricuspid bioprostheses in this anomaly might have better durability. METHODS: One hundred fifty-eight patients who received a primary tricuspid bioprosthesis because of tricuspid valve anatomy unsuitable for repair between April 1972 and January 1997 were reviewed. Results were analyzed and Kaplan-Meier curves were constructed to estimate patient survival and probability of remaining free of reoperation. RESULTS: Follow-up of 149 patients (94.3%) who survived 30 days ranged up to 17.8 years (mean, 4.5 years). Ten-year survival was 92.5%+/-2.5% (SE), 129 late survivors (92.1%) were in New York Heart Association class I or II, and 93.6% were free of anticoagulation. Freedom from bioprosthesis replacement was 97.5%+/-1.9% at 5 years and 80.6%+/-7.6% at 10 and 15 years. CONCLUSIONS: Bioprosthesis durability in the tricuspid position in patients with Ebstein's anomaly compares very favorably with bioprosthesis durability in other cardiac valve positions, especially for pediatric patients, and also compares favorably with tricuspid bioprosthesis durability in patients with other diagnoses. PMID- 9875749 TI - Alternative approach to the repair of Ebstein's malformation: intracardiac repair with ventricular unloading. AB - BACKGROUND: Moderate to severe Ebstein's malformation remains a surgical challenge. Although the various approaches that have been used are appropriate and successful in many patients, there are many for which these approaches are suboptimal. To improve the prognosis for patients across the full spectrum of Ebstein's malformation, alternative surgical approaches are necessary. METHODS: From December 1995 to October 1997, 10 patients (median age, 9 years) with moderate or severe Ebstein's malformation and mild to severe tricuspid regurgitation had partial biventricular repair with reduction of right ventricular volume load. All patients were symptomatic in New York Heart Association functional class II (n = 9) or III (n = 1). In addition to bidirectional cavopulmonary anastomosis and closure of intracardiac defects in all patients, 6 underwent tricuspid valve repair using a variety of procedures, most often simple horizontal annuloplasty. RESULTS: There were no deaths. Early reoperation was required in 1 patient (atrial septostomy on the day after operation for right ventricular failure) and another required revision of the tricuspid valve repair 10 months postoperatively for recurrent regurgitation. At follow-up ranging from 2 to 24 months, all patients are in New York Heart Association class I and have trivial tricuspid regurgitation, including the 4 who had no tricuspid valvuloplasty performed. CONCLUSIONS: We have presented an alternative approach to the management of severe Ebstein's malformation that focuses on both the tricuspid valve and the right ventricle. Just as tricuspid valve repair and reduction of regurgitation will likely improve right ventricular performance, reducing the volume load on the ventricle may improve both ventricular (right and left) and tricuspid valve function. All patients have demonstrated improved exercise tolerance and right heart function at follow-up ranging to 24 months. Additional experience will be necessary to evaluate this strategy more completely. PMID- 9875750 TI - Surgical repair of the congenitally malformed mitral valve in infants and children. AB - BACKGROUND: Mitral valve remodeling techniques were applied to 26 infants and children (mean age, 6.0 years, range, 0.4 to 15.9 years) with various forms of congenital mitral valve disease over a 7-year period. Patients with atrioventricular canal, L-transposition and single ventricle were excluded. Intraoperative transesophageal echocardiography (TEE) was utilized to assess the repair and guide the need for immediate intervention. METHODS: Twenty-one patients had mitral regurgitation: 10 with cleft anterior mitral leaflet, 7 with annular dilatation, 1 with normal leaflets with an obstructing cord, 2 with prolapsed leaflets and elongated cords, and 1 with restricted leaflet motion, normal papillary muscles, and shortened cords. Of the 5 mitral stenosis patients, 3 had supravalvular mitral ring, 1 had midvalvular mitral ring, and 1 had a parachute valve. Three of the mitral stenosis patients had additional stenotic lesions. Concurrent repair of associated lesions was performed in 21 patients (78%). RESULTS: Operative mortality was 3.8% (n = 1). There were no late deaths. Immediate rerepair in 4 patients resulted in improved function. All mitral stenosis patients improved. A total of 20 mitral regurgitation patients (95%) improved; 1 required mitral valve replacement. Mean follow-up is 31 months (range, 2 to 81 months). All patients are in New York Heart Association functional class I or II. CONCLUSIONS: Mitral valve repair can be successfully performed in infants and children with excellent short- and midterm results. Assessment using transesophageal echocardiography can guide the necessity for immediate rerepair to achieve improved function. PMID- 9875751 TI - Surgical indication for congenital heart disease with extremely thickened media of small pulmonary arteries. AB - BACKGROUND: Nineteen patients (mean age, 7.6 months) with a percent wall thickness of more than 33% in the small pulmonary arteries were found to have extremely thickened media. Based on our findings, a criterion of operative indication is proposed. METHODS: The percentage of extremely thickened media of small pulmonary arteries for all pulmonary arteries was determined on microscopic lung sections and was introduced as an index for operative indication. RESULTS: Operative repair was performed in 16 patients: 9 died intraoperatively and 7 survived more than 12 months. In 4 of 5 patients that had pulmonary artery banding, medial hypertrophy remained despite pulmonary artery banding. Operative repair also had no positive effect. In operative and late deaths and in survivors without a decrease of pulmonary arterial pressure, the percentage of extremely thickened media of small pulmonary arteries was shown to be more than 10%, whereas in 5 survivors and 1 operative death with a significant postoperative decrease of pulmonary arterial pressure, the value was less than 7%. CONCLUSIONS: If a patient has less than 7% of small pulmonary arteries with extremely thickened media, operative repair is likely to be effective. When the value is higher than 10%, not only operative repair but also pulmonary artery banding cannot be recommended because of ineffectiveness and hazard. PMID- 9875752 TI - Inoperable pulmonary vascular disease in infants with congenital heart disease. AB - BACKGROUND: Among 120 infants less than 12 months of age who had lung biopsy and autopsy, 20 were inoperable because of severe irreversible pulmonary vascular disease. METHODS: The infants were classified into three groups. Group 1 comprised 6 patients who showed complete obstruction of the small pulmonary arterial lumen and atrophy of the peripheral arterial media and who were considered to have absolute operative contraindications. Group 2 comprised 6 patients who had no pathologic findings of absolute operative contraindication and had an index of pulmonary vascular disease of more than 2.2. They were isolated as having advanced plexogenic pulmonary arteriopathy. Group 3 comprised 8 patients who had extremely thickened media of small pulmonary arteries, with abnormally thickened media extending into the small peripheral arteries characterized by extremely narrow lumina and medial thickness exceeding luminal diameter. RESULTS: Six of the 9 patients in whom operative repair was abandoned on the basis of preoperative or intraoperative lung biopsy are still alive. Of the 11 patients who underwent operation without biopsy, none survived. CONCLUSIONS: Preoperative or intraoperative lung biopsy and assessment of arteriopathy based on the above criteria are recommended in all patients in whom fatal pulmonary vascular disease is suspected. PMID- 9875753 TI - Surgical management of isolated congenital tricuspid regurgitation. AB - BACKGROUND: Isolated congenital tricuspid regurgitation without downward displacement of the leaflet is a rare clinical entity. Degenerative cusps and lack of chordae may preclude valvuloplasty and require valve replacement. METHODS: Three consecutive patients with isolated congenital tricuspid regurgitation underwent surgical repair between May 1995 and April 1997. Their ages were 23, 15, and 8 years old. Tricuspid valvuloplasty was feasible in all of them, with use of a gathering suture of the anterior leaflet, artificial chordae implantation, and ring annuloplasty. RESULTS: All 3 patients survived and recovered well after the operation. The cardiothoracic ratios on their chest roentgenograms decreased from 0.64 to 0.52 in patient 1, from 0.58 to 0.48 in patient 2, and from 0.60 to 0.44 in patient 3. Postoperative echocardiograms showed competent tricuspid valves and the disappearance of regurgitation in all cases. CONCLUSIONS: Although malformation of the valve is extensive in isolated congenital tricuspid regurgitation, application of artificial chordae with conventional valvuloplasty technique can avoid the use of prosthetic valves by establishing the competence of the tricuspid valve. PMID- 9875754 TI - Risks of repeat sternotomy in pediatric cardiac operations. AB - BACKGROUND: Repeat sternotomy has been associated with increased perioperative risks. METHODS: We reviewed 165 patients undergoing 192 repeat sternotomies between January 1985 and January 1997 (group 1) and a control group of 184 patients (group 2). The operations in group 1 were valve procedures in 94 patients, Fontan procedure in 46, ventricular septal defect closure in 10, pulmonary arterioplasty in 17, and others in 25; in group 2 ventricular or atrial septal defect closure in 120 patients, tetralogy of Fallot repair in 26, valve procedures in 16, bidirectional Glenn anastomosis in 7, repair of transposition of the great arteries in 7, pulmonary arterioplasty in 4, and others in 4. RESULTS: The hospital mortality was 2.6% in group 1 and 3.8% in group 2. Cardiac laceration occurred in 10 of 192 patients (5.2%), requiring emergent femorofemoral bypass in 6 patients. Two patients sustained an air embolism that was successfully treated with a hyperbaric chamber. Median total blood loss and requirements were not significantly different between the two groups. The length of stay in the intensive care unit and in the hospital were 4 days (range, 1 to 80 days) and 11 days (range, 1 to 135 days) in group 1, and 2 days (range, 1 to 87 days) and 7 days (range, 1 to 131 days) in group 2 (p < 0.02 and p < 0.002, respectively). The rate of complications was not significantly different in group 1 versus group 2. Overall survival was 97% (group 1) and 95% (group 2) at 120 months' follow-up (not significant). CONCLUSIONS: With careful surgical technique and judicious use of femorofemoral bypass, the risk of repeat sternotomy is minimized. PMID- 9875755 TI - Degenerative mitral regurgitation: when should we operate? AB - BACKGROUND: Left untreated, severe mitral regurgitation in asymptomatic patients can lead to irreversible cardiac damage, which can develop with little warning. Over the period of this study, we have tended to operate earlier in the disease process and on less symptomatic patients. We report here our experience. METHODS: Between January 1985 and June 1996, 710 patients with mitral regurgitation underwent operations. Three hundred twenty-nine (213 male and 116 female with a mean age of 65.5 years) had degenerative mitral valve disease and of this group, 169 patients underwent repair and 160, replacement. RESULTS: The overall operative mortality was 4 patients (1.2%). There were no operative deaths among patients having isolated mitral valve repair. Survival at 1 year, 5 years, and 10 years was 94%+/-1.4% (+/- the standard error of the mean), 77%+/-2.9%, and 41%+/ 5.8%, respectively. Survival was significantly better in the group having repair (p < 0.05). Ten patients (6%) in the repair group and 13 (8%) in the replacement group required reoperation. Increased age, worse left ventricular function, type of operation (replacement worse than repair), and increased left ventricular size were significantly associated with poorer survival. CONCLUSIONS: These data confirm the superior results achieved with mitral valve repair and support early mitral valve repair before functional deterioration. PMID- 9875756 TI - Determinants of midterm outcome of partial left ventriculectomy in dilated cardiomyopathy. AB - BACKGROUND: Partial left ventriculectomy has been proposed for treatment of severe cardiomyopathies. This study reports midterm results of this procedure in 37 patients with dilated cardiomyopathy. METHODS: All patients were in New York Heart Association class III (16) or IV (21). Partial ventriculectomy was associated with mitral annuloplasty in 27 patients and with mitral replacement in 2. RESULTS: There were seven operative deaths (18.9%). During a mean follow-up of 18.2+/-9.3 months, 9 more patients died. Actuarial survival was 56.7%+/-8.1% at 6 and 24 months. Analysis of factors influencing outcome showed that midterm survival was significantly affected only by myocardial cell diameter. Otherwise, functional class improved from 3.5+/-0.5 to 1.8+/-0.9 in the survivors (p < 0.001). Furthermore, left ventricular diastolic volume decreased from 523+/-207 to 380+/-148 mL (p < 0.001), and left ventricular ejection fraction increased from 17.1%+/-4.6% to 23%+/-8% (p < 0.001), whereas significant changes in cardiac index, stroke index, and pulmonary pressures were found at 1 month of follow-up. Although left ventricular diastolic volume tended to increase in the late postoperative period, left ventricular ejection fraction and hemodynamic variables did not change significantly. CONCLUSIONS: Partial ventriculectomy improves left ventricular function and congestive heart failure in patients with dilated cardiomyopathy for up to 24 months of follow-up. Nevertheless, this procedure's clinical application is limited by the high mortality observed in the first postoperative months. Otherwise, new perspectives may be advised by the identification that partial ventriculectomy results seem to be influenced by compromised myocardial cells. PMID- 9875757 TI - Disadvantages of local repair in acute type A aortic dissection. AB - BACKGROUND: In acute type A dissection of the aorta, local repair with glue aortoplasty was compared with aortic replacement. METHODS: Between 1992 and 1996, 106 consecutive patients (mean age, 59 years; 84 men) were operated on average 14.5 hours after onset of dissection. A local repair (gelatin-resorcine formaldehyde/glutaraldehyde glue, Trigon AG, Monchengladbach, Germany) without graft replacement was performed in 21 patients. Graft replacement and reinforcement of aortic stumps with gelatin-resorcine-formaldehyde/glutaraldehyde glue was performed in 85 patients (supracoronary graft, 68; aortic root replacement, 17). RESULTS: Survival was 79% after 30 days and 69% after 2 years. There was no difference in early mortality (p = 0.2240) and survival (p = 0.07649). Risk factors for early mortality were preoperative shock, neurologic disorder, duration of crossclamp, and extracorporeal circulation. The rate of reoperation on the proximal aorta was 31.6% (6 of 19) after local repair and 9% (6 of 64) after aortic replacement (p = 0.0157). Local repair was a significant predictor for reoperation (p = 0.0087), with decreased reoperation-free survival (p = 0.01164). In all reinterventions (four supracoronary grafts, including two valve replacements; two composite grafts; two arch replacements) breakdown of the aortoplasty was confirmed. CONCLUSION: Local repair has satisfactory early results but an increased incidence of reoperations due to a breakdown of the glue aortoplasty. Indications for local repair should be restricted to high-risk patients requiring a minimal emergency surgical procedure. PMID- 9875758 TI - Carnitine supplementation improves myocardial function in hearts from ischemic diabetic and euglycemic rats. AB - BACKGROUND: Nonischemic myocardial dysfunction in patients with diabetes mellitus appears to be attenuated with long-term L-carnitine therapy. The effect of acute L-carnitine supplementation on rat hearts from euglycemic and diabetic animals subjected to ischemia and reperfusion is investigated in this study. METHODS: Study rats had diabetes mellitus induced by streptozocin (65 mg/kg intraperitoneally), and control rats had injection of saline solution (n = 12 per group). About 1 month later, the hearts were suspended on a Langendorff apparatus and perfused with either standard buffered Krebs-Henseleit solution or this standard solution supplemented with L-carnitine (5 mmol/L). After stabilization, normothermic, zero-flow ischemia was instituted for 20 minutes followed by 60 minutes of reperfusion. There were four study groups (n = 6 per group): hearts that were from euglycemic rats and that were perfused with standard buffered Krebs-Henseleit solution (E-STD); hearts that were from diabetic animals and that were perfused with the same standard buffered solution (DM-STD); hearts taken from diabetic animals and perfused with L-carnitine-enriched solution (DM-CAR); and hearts taken from euglycemic rats and perfused with the enriched solution (E CAR). RESULTS: At 60 minutes of reperfusion, left ventricular developed pressure was significantly better in hearts from both groups (diabetic and euglycemic) with carnitine supplementation (DM-CAR versus DM-STD and E-CAR versus E-STD, p < 0.01 for both, by analysis of variance). Left ventricular end-diastolic pressure was significantly lower in the DM-CAR group compared with all other groups (p < 0.01 by analysis of variance). CONCLUSIONS: These findings suggest that acute L carnitine supplementation significantly improves the recovery of the ischemic myocardium in diabetic and euglycemic rats. PMID- 9875759 TI - Repair of ruptured sinus of valsalva aneurysm: determinants of long-term survival. AB - BACKGROUND: Ruptured sinus of Valsalva aneurysm is a rare cardiac anomaly and long-term survival after surgical treatment is not well established. This study was designed to investigate the determinants of long-term survival after repair of ruptured sinus of Valsalva aneurysm. METHODS: From April 1978 to April 1996, 53 patients underwent operation for ruptured sinus of Valsalva aneurysm. The incidence among our cardiac surgical population was 0.56%. Long-term survival was investigated in 46 patients (13 to 65 years) who survived the operation, with 96.2% follow-up completeness (mean+/-standard deviation, 6.5+/-4.9 years; maximum, 17.2 years), by univariate and multivariate analyses. RESULTS: There was no early operative death and no recurrence after the initial repair. Actuarial survival was 83.8%+/-8.4% at 15 years. Reoperation, aneurysm draining into the left ventricle, aortic prosthetic dehiscence, bacterial endocarditis, and aortic cross-clamp time (<70 minutes) were significant factors in long-term survival (p < 0.05). Multivariate analysis revealed that only aortic prosthesis dehiscence was the significant factor influencing late survival (p = 0.0001). CONCLUSIONS: Surgical treatment for ruptured sinus of Valsalva aneurysm is safe and has satisfactory results. Aortic prosthesis dehiscence is the independent determinant for long-term survival. Other factors including bacterial endocarditis, concomitant ventricular septal defect repair, and aortic valve replacement did not independently influence long-term survival. PMID- 9875760 TI - Neuropsychological dysfunction after minimally invasive direct coronary artery bypass grafting. AB - BACKGROUND: We compared postoperative neuropsychological dysfunction after minimally invasive direct coronary artery bypass grafting (MIDCAB) operation with coronary artery bypass graft operations using cardiopulmonary bypass. METHODS: Neuropsychological assessment was performed preoperatively and before discharge on 7 patients undergoing MIDCAB procedures, 9 patients undergoing single-graft cardiopulmonary bypass operation, and 27 patients undergoing multiple-graft cardiopulmonary bypass operation. From a matched control group of 40 normal subjects reliable change indices were derived for each measure and used to determine the incidence of postoperative decline. RESULTS: There was little difference between the MIDCAB and single-graft cardiopulmonary bypass groups on the incidence of neuropsychologic decline. However, the multiple-graft cardiopulmonary bypass group had a significantly higher incidence of decline than the MIDCAB and single-graft cardiopulmonary bypass groups on specific neuropsychologic measures, coupled with a significantly greater number of postoperative deteriorations per patient. CONCLUSIONS: The elimination of cardiopulmonary bypass does not prevent neuropsychological dysfunction after cardiac operation as patients undergoing MIDCAB and single-graft cardiopulmonary bypass experience similar deteriorations in performance. However, the deterioration is markedly worsened when the number of surgical grafts is increased. PMID- 9875761 TI - Triiodothyronine reverses depressed contractile performance after excessive catecholamine stimulation. AB - BACKGROUND: Conflicting results have been reported regarding the acute effects of triiodothyronine (T3) on myocardial contractile performance. The present study analyzes the role of T3 in reversing the depressant effect of excessive catecholamine stimulation in isolated porcine left ventricular myocardium. METHODS: Thirty-six left ventricular trabeculae (0.4 x 6.0 mm) obtained from 6 pigs were used for measurements of isometric force development, isotonic shortening, and intracellular calcium in three experimental series (measurement conditions: 37 degrees C; optimal length; supramaximal electrical stimulation, 1 Hz; calcium measurement, fura-2 ratio method; frequency, 225 Hz). In series 1, isometric force development was measured before and after a 60-minute incubation with 10(-7) mol/L epinephrine in preparations with (n = 6) and without (n = 6) preceding fura-2 loading for calcium measurements. In series 2, the acute effects of a 30-minute administration of T3 (10(-9) mol/L) on isometric force and intracellular calcium were analyzed (n = 6). In series 3, after simultaneous fura 2 loading and a 6-hour 10(-7) mol/L epinephrine exposure the effects of T3 (10( 9) mol/L, 30 minutes) on force development, shortening, and intracellular calcium transient were analyzed. RESULTS: Long-term and high-dose epinephrine exposure induced a severe contractile depression with a significant reduction of isometric force development (p < 0.05) and increased diastolic (p < 0.001) and systolic calcium (p < 0.001). In normal porcine myocardium T3 had no effect on the extent of isometric force generation but accelerated the time course of force development (p < 0.05) and increased the calcium transient (p < 0.001). After induction of myocardial depression by epinephrine exposure T3 accelerated the intracellular calcium transients and reduced diastolic calcium. Triiodothyronine increased the shortening amplitude and the force amplitude (p < 0.01). CONCLUSIONS: Triiodothyronine reverses depressed contractile performance after preceding high-dose epinephrine exposure in isolated porcine myocardium. Increased force amplitudes and unaltered or even reduced intracellular calcium transients argue in favor of a resensitization of the contractile apparatus for calcium by T3. The study supports a potential role for T3 treatment in depressed myocardium after previous excessive catecholamine exposure (eg, brain death, catecholamine treatment, ischemia). PMID- 9875762 TI - Clinical experience with the video-assisted saphenectomy procedure for coronary bypass operations. AB - BACKGROUND: Leg wound complications after saphenectomy are frequent after coronary bypass operations and have a detrimental effect on postoperative quality of life and treatment cost. To reduce morbidity, we evaluated a new technique of video-assisted vein harvest. METHODS: Between March 1996 and October 1996, 50 patients had video-assisted saphenectomy (VAS) and 40 patients had the standard open technique (control group). An additional 13 patients had both procedures (hybrid group). Level of pain, edema, and wound complications were evaluated at discharge and at 2, 4, and 6 weeks postoperatively. RESULTS: The mean operating time for VAS patients was slightly higher than for control (60.6+/-24.7 minutes versus 53.2+/-21.1 minutes; p > 0.05). The average incision length in VAS patients was 13.8+/-8.8 cm for an average of 3.3 grafts per patient. Three VAS procedures were aborted, two because of time constraints, and one because of bleeding, and a segment of vein was lost to injury. The VAS group had considerably less early postoperative pain than the control group (1.7+/-1.2 versus 4.1+/-1.4 [1 = mild, 10 = severe]; p < 0.005) and edema was similar for both groups. Patients in the hybrid group reported less pain in the VAS-operated leg. Serious wound infection occurred in 4 patients, with 2 patients in the control group requiring reoperation for drainage and flap reconstruction. CONCLUSIONS: Based on this initial experience, VAS harvesting, although initially more time consuming, is a rapidly mastered technique, results in shorter overall incision length, and is associated with considerably less postoperative pain than the standard open technique. PMID- 9875763 TI - Coronary artery bypass grafting in patients with advanced left ventricular dysfunction. AB - OBJECTIVE: The aim of this study was to determine the long-term survival and control of angina in patients with coronary artery disease and sequentially decreased ejection fractions (EF) after first-time coronary artery bypass grafting. METHODS: Between 1981 and 1995, 156 (1.3%) patients with an EF less than 0.25 (group 1), 588 (5%) patients with an EF of 0.25 to 0.34 (group 2), 2,438 (20.6%) patients with an EF of 0.35 to 0.49 (group 3), and 8,648 (73.1%) patients with an EF equal to or greater than 0.50 (group 4) underwent coronary artery bypass grafting. The EFs were determined by uniplanar or biplanar left ventriculography. For each group, the clinical and angiographic characteristics and the operative and outcome data were compared. Survival curves were derived and compared for each group. Correlates of angina, and of early (30-day) and long term mortality, for all groups were analyzed. RESULTS: For all groups the mean age was approximately 60+/-10 years. Group 1 had the highest percentage of patients who were men (88%), had congestive heart failure (34%), had hypertension (53%), and had left main coronary artery disease (24%). Groups 1 through 3, compared with group 4, had a lower percentage of complete revascularization (p < 0.0001), a lower percentage of internal mammary artery grafts (p < 0.0001), and a greater use of intraaortic balloon pump (p < 0.0001), but had similar cross-clamp and cardiopulmonary bypass times, number of grafts, incidences of myocardial infarction, and stroke. Hospital mortality for groups 1, 2, 3, and 4 was 3.8% (n = 6), 3.4% (n = 20), 3% (n = 72), and 1.6% (n = 134), respectively. Groups 1 through 3, compared with group 4, had similar incidences of angina during follow up (31% to 40% versus 33%, respectively; p < 0.06). Survival was greatest for group 4 compared with groups 1 through 3 at 1, 5, and 10 years (p < 0.0001). Patients in group 1 had 1-, 5-, and 7-year survivals of 90%, 64%, and 49%. Multivariate correlates of early mortality were advanced age, female sex, decreased EF, hypertension, diabetes, and emergency operation. Multivariate correlates of long-term mortality included severity of preoperative angina class, congestive heart failure, number of diseased vessels, and incomplete revascularization. The strongest correlates of angina at follow-up were younger age, female sex, previous myocardial infarction, lower ejection fraction, and incomplete revascularization. The absence of an internal mammary artery graft did not predict the occurrence of angina or influence long-term survival. CONCLUSIONS: In the long term there is a higher mortality in patients with sequentially decreased left ventricular function undergoing coronary artery bypass grafting, although more than 60% of patients with an EF less than 0.25 were alive and had good control of angina at 5 years despite having a higher percentage of risk factors, poorer functional status, and more complex coronary disease. Failure of symptom control and survival beyond 5 years appeared to be influenced by preexisting medical conditions and factors that affect the ability to completely revascularize the myocardium. These results suggest that in selected patients with ischemia and poor left ventricular function, coronary artery bypass grafting may preserve remaining viable myocardium, provide relief of symptoms, and offer survival greater than 60% at more than 5 years. PMID- 9875764 TI - "Bow-tie" mitral valve repair: an adjuvant technique for ischemic mitral regurgitation. AB - BACKGROUND: Current techniques of mitral valve repair rely on decreasing valve area to increase leaflet apposition, but fail to address subvalvular dysfunction. A novel repair has been introduced with partial left ventriculectomy, which apposes the anterior leaflet to a corresponding point on the posterior leaflet creating a double-orifice valve, with reported adequate control of mitral regurgitation. METHODS: We started to use the "bow-tie" repair as an adjunct to posterior ring annuloplasty in cases in which mitral regurgitation was not adequately controlled by decreasing mitral valve area (n = 6), or when placement of an annuloplasty ring was impractical (n = 4). Mean follow-up was 336 days (range, 82 to 551 days) with no postoperative deaths. RESULTS: Mitral regurgitation decreased from 3.6+/-0.5 to 0.8+/-0.4 (p < 0.0001), with a concomitant increase in ejection fraction from 33%+/-13% to 45%+/-11% (p = 0.0156) before hospital discharge. Mitral valve area, measured by pressure half time, decreased from a mean of 2.5+/-0.3 to 2.1+/-0.3 cm2, with a mean transvalvular gradient of 4.5+/-2.0 mm Hg. In patients whose mitral valve was repaired using the bow-tie alone, mitral regurgitation was reduced from 4+, to a trace to 1+. Postoperatively, mitral valve area increased from 1.9 to 2.5 cm2 during exercise, further supporting the concept that this technique preserves mitral valve annular function. CONCLUSIONS: These observations suggest that the bow-tie repair may offer advantages over conventional techniques of mitral valve repair and should be considered as an adjunct, especially in patients with impaired left ventricular function. PMID- 9875765 TI - Lazaroid U74500A is superior to U74006F in preserving rabbit heart for 24 hours. AB - BACKGROUND: Lazaroid, a series of 21-aminosteroids, has been shown to reduce free radical-mediated injury after ischemia and reperfusion. Recent in vitro studies have demonstrated that, among the various compounds studied, the most efficient agent was U74500A. The question is whether these findings apply to the whole heart experiencing ischemia-reperfusion injury. In this study we compared the myocardial protective effects of U74006F, the only clinical candidate, and U74500A. METHODS: An isolated rabbit heart preparation perfused with the blood from a support rabbit was used. All hearts were divided into three groups according to the administration of U74500A (4 mg/kg, group A; n = 7), U74006F (4 mg/kg, group F; n = 7), or solvent (group S; n = 7) to the donor rabbit before preservation. After 24 hours of preservation with University of Wisconsin solution at 0 degrees C, all hearts were perfused with cross-circulated blood for 60 minutes with the Langendorff mode followed by 40 minutes in the working mode. RESULTS: After 10 minutes of reperfusion the serum lipid peroxide levels were significantly (p < 0.05) lower in group A (0.62+/-0.31 nmol/mL) than those in group S (2.1+/-1.3 nmol/mL) and group F (1.0+/-0.6 nmol/mL). The aortic flow rate at 10 mm Hg of left atrial pressure was significantly higher in group A (164+/-37 mL/min) than that of other groups (71+/-28 mL/min in group S and 97+/-28 mL/min in group F). There were no significant differences in high-energy phosphate levels after reperfusion among the three groups. CONCLUSION: These data imply that U74500A inhibits lipid peroxidation and prevents ischemia-reperfusion injury more efficiently than U74006F. PMID- 9875766 TI - Fluorescence activated cell sorting: a reliable method in tissue engineering of a bioprosthetic heart valve. AB - BACKGROUND: Techniques of tissue engineering are used to seed human autologous cells in vitro on degradable mesh to create new functional tissue like a bioprosthetic heart valve. A precondition is subsequent seeding of native-valve analogous pure endothelial and myofibroblast cell lines. The aim of this study is to find a safe method of isolating viable cell lines out of tissues from the operating room. METHODS: Mixed cells from ascending aorta obtained from the operating room were incubated with an endothelial-specific fluorescent marker. The labeled cells were activated and sorted by flow cytometry. Isolated cell lines were cultured and thereafter square sheets of polymeric scaffold were seeded with myofibroblasts, followed by endothelial cells. The created tissue was stained with hematoxylin and eosin, van Gieson stain, and stains for factor VIII and CD34. RESULTS: Control culture samples (n = 25) revealed vital uncontaminated endothelial and myofibroblast cell lines. Microscopy of the seeded meshes (n = 16) demonstrated a tissue-like structure. Van Gieson stain showed production of collagen. Endothelial cells formed a superficial monolayer, demonstrated by factor VIII and CD34; no invasive formation of capillaries was detectable. CONCLUSIONS: These results demonstrate that fluorescence activated cell sorting is a reliable and safe method to gain pure vital autologous cell lines out of human mixed cells for subsequent seeding on degradable mesh and that those cells are active to form new tissue. PMID- 9875767 TI - Protective action of 17beta-estradiol in cardiac cells: implications for hyperkalemic cardioplegia. AB - BACKGROUND: Hyperkalemic cardioplegic solutions effectively arrest the heart, but may also induce intracellular Ca2+ loading and cellular hypercontracture, which could contribute to ventricular dysfunction associated with global surgical ischemia. Recently, it has been proposed that 17beta-estradiol may possess protective properties in the ischemic myocardium. The purpose of the present study was to examine the action of 17beta-estradiol on cardiac cells exposed to hyperkalemic stress. METHODS: Single ventricular cardiomyocytes, a preparation devoid of vascular and neuronal elements, were isolated from guinea pig hearts, loaded with a Ca2+-sensitive fluorescent probe, and imaged by digital epifluorescent microscopy. The emitted fluorescence of the probe, a measure of intracellular Ca2+ concentration, and cell length were simultaneously recorded during hyperkalemic challenge, in the absence or presence of 17beta-estradiol. RESULTS: In control cardiomyocytes, the cytosolic concentration of Ca2+ was 138+/ 11 nmol/L and cell length 93+/-11 microm. Exposure to high K+ (+16 mmol/L KCl) significantly increased cytosolic Ca2+ to 2,191+/-87 nmol/L (p < 0.001), and produced cell shortening (length at 39+/-5 microm; p < 0.001). 17beta-Estradiol (10 micromol/L) acutely prevented high K+ to induce either intracellular Ca2+ loading (144+/-13 nmol/L, p < 0.001) or hypercontracture (91+/-10 microm, p < 0.001). Tamoxifen (10 micromol/L), an antiestrogen, abolished the protective effect of 17beta-estradiol. CONCLUSIONS: We conclude that 17beta-estradiol prevents hyperkalemia-induced Ca2+ loading and hypercontracture through a direct and tamoxifen-sensitive action in cardiomyocytes. This study raises the possibility that 17beta-estradiol should be considered as a cardioprotective adjunct toward a safer hyperkalemic cardioplegia. PMID- 9875768 TI - The CardioWest total artificial heart bridge to transplantation: 1993 to 1996 national trial. AB - BACKGROUND: We performed a controlled study of a total artificial heart in bridge to transplantation. We hypothesized that the CardioWest total artificial heart used in a selected population of decompensating cardiac transplantation candidates would result in improved survival compared with matched controls. METHODS: The CardioWest trial started in 1993 in six United States institutions under an investigational device exemption from the Food and Drug Administration. Four centers contributed 27 implant and 18 matched retrospective control patients. RESULTS: Of the implant patients, 25 (93%) received a transplant, 24 (89% of the total, 96% of those transplanted) were discharged and are currently surviving. In the control group, 10 patients died awaiting transplantation, 8 received a transplant, and 7 were discharged with 6 surviving (p = 0.00001). All adverse events were documented with respect to time. Thirteen serious adverse events occurred, 11 of which occurred in the 2 patients that died during implant. CONCLUSIONS: In a selected group of patients with endstage heart disease, use of the CardioWest total artificial heart is lifesaving. When compared with the series of matched retrospective controls, a significant improvement in survival was found in the CardioWest implant group. PMID- 9875769 TI - Cardiac operations in the elderly: who is at risk? AB - BACKGROUND: With the growing number of elderly patients presenting for cardiac operations we analyzed their early survival data to determine whether any preoperative variables might be indicative of increased risk. METHODS: From 1990 to 1995, 436 consecutive patients who were 75 years old or older had either coronary artery bypass, valve replacement(s), or a combination of these. A total of 34 preoperative variables were assessed for their effect on hospital survival by using univariate and multivariable analysis. RESULTS: There were 266 men and 170 women, with 292 patients being 75 to 80 years old and 144 patients being older than 80 years. Coronary artery bypass was performed in 242 patients, valve replacement was performed in 93 patients, and a combination of these in 101 patients. The operation was considered elective in 202 patients, urgent in 209, and emergent in 25 patients of whom 21 were in cardiogenic shock. Overall there were 61 hospital deaths (13.9%). The most common cause of death, low cardiac output syndrome, occurred in 34 patients of whom 26 suffered a perioperative myocardial infarction. Stroke was the cause of death in eight and multiple organ failure accounted for nine deaths. In the univariate analysis, variables that influenced survival included heart failure (p = 0.004), pulmonary edema (p = 0.004), cardiomegaly (p = 0.02), elevated serum creatinine (p = 0.009), surgical priority (p = 0.002), and cardiogenic shock (p = 0.002). In the multivariable analysis there were three independent determinants of hospital survival: cardiomegaly (odds ratio, 1.8:1) serum creatinine level higher than 150 micromol/L (odds ratio, 5.5:1) and emergency procedure (odds ratio, 2.5:1). CONCLUSIONS: Although cardiac operations can be performed safely in many elderly patients, we identified several factors that might help both in case selection and in perioperative decisions. PMID- 9875770 TI - Pattern and significance of cerebral microemboli during coronary artery bypass grafting. AB - BACKGROUND: Strokes that occur during coronary artery bypass grafting are often caused by embolism. Intraoperative transcranial Doppler monitoring can detect cerebral microemboli. The aims of this study were to identify the pattern of microembolic phenomena during various stages of coronary artery bypass grafting, to verify whether numbers of high-intensity transient signals correlated with early neuropsychologic deficits, and to identify, using magnetic resonance imaging scans, whether radiologic evidence of cerebral infarction correlated with microembolic numbers during the bypass period. METHODS: Forty-one consecutive patients undergoing coronary bypass grafting with transcranial Doppler monitoring were enrolled in this study. All had preoperative and postoperative magnetic resonance imaging brain scans. A subgroup of 32 patients were studied by comparing microembolic load and early neuropsychological outcomes. RESULTS: Transcranial Doppler monitoring confirmed that most microemboli occurred during cardiopulmonary bypass. A significant early neuropsychological deficit after coronary artery bypass grafting did correspond to the total microembolic load during bypass (p = 0.008). However, patients with cerebral infarction on magnetic resonance imaging had significantly more microembolic signal during the preincision phases and not during the bypass period. CONCLUSIONS: Microembolic load during bypass is associated with early neuropsychologic deficits. In contrast, patients who show evidence of strokes during coronary artery bypass grafting have a higher microembolic load during the preincision phase than those without cerebral infarction. Differing mechanisms may be responsible for these different outcomes. PMID- 9875771 TI - Experimental confirmation of effectiveness of fenestration in acute aortic dissection. AB - BACKGROUND: Aortic fenestration is used clinically to treat organ ischemia in acute descending aortic dissection. However, fenestration has not been studied experimentally. This study does so using an animal model. METHODS: Descending aortic dissection was created in six dogs, with subsequent fenestration of the infrarenal aorta. Blood flow (femoral, cephalic, and renal), blood pressure (femoral and carotid), and aortic distensibility were measured at baseline, after dissection, and after fenestration. Values were compared using paired t tests. RESULTS: Baseline femoral, cephalic, and renal arterial flows were 53+/-37, 78+/ 65, and 83+/-52 mL/min, respectively. Baseline femoral and carotid pressures were 82+/-13 and 81+/-11 mm Hg, respectively. After dissection, femoral, cephalic, and renal arterial flow decreased to 20+/-21 (p < 0.05), 38+/-26, and 56+/-36 mL/min, respectively. Femoral blood pressure decreased to 28+/-17 mm Hg (p < 0.05). With fenestration, femoral, cephalic, and renal flows increased to 60+/-37 (p < 0.05), 78+/-51, and 80+/-48 mL/min, respectively. Femoral blood pressure increased to 85+/-28 mm Hg (p < 0.05). Carotid pressure remained unchanged with dissection and fenestration (77+/-17 mm Hg, 82+/-17 mm Hg, respectively). Baseline aortic distensibility (21%) decreased significantly after dissection (to 1.4%, p < 0.05) and increased after fenestration (to 12%, p < 0.05). CONCLUSIONS: Experimental aortic fenestration restored blood pressure and flow to hypoperfused organs in acute descending aortic dissection. The continued clinical application of fenestration is supported. PMID- 9875772 TI - Outcome analysis of 245 CarboMedics and St. Jude valves implanted at the same institution. AB - BACKGROUND: Thromboembolism and valve-related death are major complications associated with prosthetic valve implants, but it is difficult to evaluate the relative incidence of these complications based on studies in which the implantation of only one valve is reported from any given institution. We therefore report the outcome of patients implanted at our institution during the same time period with either the recently released CarboMedics (CM) or the St. Jude Medical (SJ) valve prostheses. METHODS: Between October 1994 and January 1996, 245 consecutive patients received either SJ (116 patients) or CM (129 patients) valves at our institution. Follow up of these patients was 99.6% complete, for a total of 318.5 cumulative patient-years (median follow-up, 1.4 years). RESULTS: The 30-day mortality rates for SJ and CM implants were 3.4% and 3.1%, respectively. Actuarial survival and freedom from valve related mortality rates at 1.5 years for SJ and CM valves were 94%+/-2% versus 86%+/-3% (p = 0.03) and 100% versus 94%+/-2% (p = 0.005), respectively. There was no structural valve failure for either implant, but there were five thrombosed valves in the CM group and none in the SJ group (p = 0.04). All thrombosed valves were mitral (four mitral valve replacement, one aortic and mitral valve replacement). Two of the thrombosed valves were successfully explanted, whereas the three remaining patients died. Freedom from a thromboembolic event in the mitral position at 1.5 years, including thrombosed valves was 97%+/-3% and 83%+/-5% for SJ and CM valves, respectively (p = 0.04). CONCLUSIONS: The results of this study suggest that further evaluation of thromboembolic outcomes after CM compared with SJ valve implantation is warranted. PMID- 9875773 TI - Flexible posterior mitral annuloplasty: five-year clinical and Doppler echocardiographic results. AB - BACKGROUND: Stabilization of the posterior annulus seems to be a critical factor to achieve a stable mitral valve repair. To assess the benefit of softer mural annuloplasty, we analyzed results obtained with the flexible linear reducer. METHODS: From 1985 to 1993, 120 patients, with pure mitral regurgitation, mainly degenerative, had a mitral reconstruction. Mean age was 64+/-11 years and 74% of the patients were in New York Heart Association functional class III or IV. RESULTS: Hospital mortality was 3.3%. Mean follow-up was 56+/-24 months. There were 23 late deaths; 10 valve-related including 7 sudden deaths. Two patients (1.7%) required a reoperation. Doppler echocardiographic studies revealed excellent valve function; 5-year freedom from significant regurgitation was 85.8%+/-5.4%. Mean mitral valve area was 2.76+/-0.77 cm2. Although 105 patients were in class I or II, 23 patients were not functionally improved. Previous myocardial infarction and shorter deceleration time of early filling were risk factors for worsening functional disability. CONCLUSIONS: This support provides stable repair with excellent clinical and echographic results. Previous myocardial infarction and noncompliant left ventricle negatively influence outcome. PMID- 9875774 TI - Permanent cardiac pacing after a cardiac operation: predicting the use of permanent pacemakers. AB - BACKGROUND: The need for permanent cardiac pacing after cardiac operations is infrequent but associated with increased morbidity and resource utilization. We identified patient risk factors for pacemaker insertion to enable development of a predictive model. METHODS: Data were collected prospectively for 10,421 consecutive patients who had cardiac operations between January 1990 and December 1995. Two hundred fifty-five patients (2.4%) were identified as having received a permanent pacemaker during the same hospitalization. Logistic regression analysis was performed to determine the independent, multivariate predictors of permanent pacing. The predictive accuracy and precision of the logistic regression model was evaluated in the 1996 database of 2,236 consecutive patients by the calculation of Brier scores. RESULTS: Eight independent predictors of permanent pacemaker requirement were identified. The factor-adjusted odds ratios (OR) with 95% confidence interval (CI) associated with each predictor are as follows: (1) valve replacement surgery (aortic: OR 5.8, CI 3.9-8.7; mitral: OR 4.9, CI 3.1 7.8; tricuspid: OR 8.0, CI 5.5-11.9; double: OR 8.9, CI 5.5-14.6; and triple: OR 7.5, CI 2.9-19.3); (2) repeat operation: OR 2.4, CI 1.8-3.3; (3) age 75 years or older: OR 3.0, CI 2.0-4.4; (4) ablative arrhythmia operation: OR 4.2, CI 1.9-9.5; (5) mitral valve annular reconstruction: OR 2.4, CI 1.4-4.2; (6) use of cold blood cardioplegia: OR 2.0, CI 1.2-3.6; (7) preoperative renal failure: OR 1.6, CI 1.0-2.6; and (8) active endocarditis: OR 1.7, CI 0.9-3.0. A model for postoperative permanent pacemaker requirement using the eight predictors was formulated and tested (Brier score = 0.017+/-0.003; Z = 0.18). CONCLUSIONS: The proposed predictive model correlated highly with actual pacemaker use, which suggests that the requirement for pacing results from either operative trauma or increased ischemic burden. Preoperative identification of patients at increased risk of conduction disturbances may allow for earlier detection and improved treatment. Patients requiring postoperative pacing had increased morbidity and length of stay. PMID- 9875775 TI - Nonpenetrating stapling: a valuable alternative for coronary anastomoses? AB - BACKGROUND: The safe development of minimally invasive coronary artery bypass operations might require alternatives to conventional suture-based anastomotic techniques. In this setting, nonpenetrating stapling is an attractive option because of its simplicity of use and potential for improved endothelial preservation. METHODS AND RESULTS: In the experimental part of this study, porcine internal mammary arteries were anastomosed to left anterior descending coronary arteries using either an 8-0 polypropylene running suture or nonpenetrating microclips (7 anastomoses in each group). The endothelium dependent relaxations to bradykinin of the arterial rings bearing the anastomosis and of noninstrumented rings were compared in organ chamber experiments. There were no significant differences in maximal relaxations (mean +/- SEM) between the microclipped and sutured anastomoses (81%+/-7% versus 74%+/-10%), which were both significantly lower than those of control coronary rings (98%+/-2%, p = 0.001 versus the two anastomosed groups). Histologic examination showed a comparable preservation of the coronary and graft endothelium with both techniques. The clinical part of the study comprised 7 patients in whom the left internal mammary artery was conventionally sutured to the left anterior descending whereas 13 saphenous vein grafts were anastomosed to their target vessels by nonpenetrating staples. There were no clip-related complications. An angiographic assessment of the venous grafts was performed within 10 days postoperatively in all patients. One graft is presumably occluded. The 12 remaining conduits were patent with stapled anastomoses featuring a widely open "shark-mouth" configuration. CONCLUSIONS: These preliminary data suggest that non-penetrating stapling is an easy-to-use technique that competes well with conventional suturing, at least in terms of immediate results. Further studies are warranted to better define its potential place within the armamentarium of minimally invasive coronary artery bypass techniques. PMID- 9875776 TI - Pulmonary metastasectomy for testicular germ cell tumors: a 28-year experience. AB - BACKGROUND: The role of surgery in patients with pulmonary metastatic germ cell tumors has been evolving since the 1970s. To evaluate the results of pulmonary resection, we reviewed our 28-year experience. METHODS: Between July 1967 and May 1995, 157 patients with testicular germ cell tumors underwent pulmonary resections for suspected metastases. Their clinical and pathological data were reviewed. Kaplan-Meier and Cox regression models were used to analyze prognostic factors for survival after resection of metastatic disease. RESULTS: All patients were male with median age of 27 years (range 15-65). Complete resection was accomplished in 155 (99%) patients. Viable carcinoma was present in 44% (70) of the patients. Forty-one (26%) patients had metastases to other sites after pulmonary metastasectomy. The overall actuarial survival 5 years after pulmonary resection was 68% for the entire group and 82% for patients diagnosed after 1985. On multivariate analysis, the adverse prognostic factors were metastases to nonpulmonary visceral sites (p = 0.0069) and the presence of viable carcinoma in the resected specimen (p < 0.0001). CONCLUSIONS: With current chemotherapy regimens, almost 85% of the patients with testicular germ cell tumors undergoing complete resection of their pulmonary metastases can be expected to achieve long term survival. PMID- 9875777 TI - Detection of micrometastases in histologically negative lymph nodes in esophageal cancer. AB - BACKGROUND: New molecular techniques may identify micrometastases in histologically negative lymph nodes and have an impact on the staging of esophageal cancer. We investigated the role of the reverse transcriptase polymerase chain reaction (RT-PCR) assay to identify micrometastases in esophageal cancer. METHODS: The RT-PCR assay to detect carcinoembryonic antigen (CEA) messenger ribonucleic acid (mRNA) was performed on lymph nodes from patients with esophageal cancer and benign esophageal disorders. The presence of CEA mRNA in lymph nodes was considered evidence of metastases. RESULTS: Histopathologic study revealed metastases in 50 (41%) of 123 lymph nodes from 30 patients with esophageal cancer. All histologically positive lymph nodes contained CEA mRNA by RT-PCR. Of 73 histologically negative lymph nodes, 36 (49%) contained CEA mRNA, a significant increase compared with the histopathologic diagnosis (p < 0.001). Lymph nodes in patients with benign disease contained no CEA mRNA. In 10 patients, histologic stage was NO. Five of them were also negative by RT-PCR, and all are alive with only one recurrence. In the remaining 5 patients, RT-PCR was positive for occult lymph node metastases; 2 have died of disease, and 1 is alive with recurrent disease. CONCLUSIONS: In patients with esophageal cancer, RT-PCR detects more lymph node metastases than does histopathology. Initial follow-up suggests a positive RT-PCR with negative histologic findings may have poor prognostic implications. Further studies will be needed to confirm any clinical implications. PMID- 9875778 TI - Isolated lung perfusion with melphalan and tumor necrosis factor for metastatic pulmonary adenocarcinoma. AB - BACKGROUND: Isolated left lung perfusion with melphalan and human tumor necrosis factor-alpha for pulmonary metastatic adenocarcinoma in the WAG/Rij rat was studied. METHODS: Survival was determined for melphalan, human tumor necrosis alpha. Lung, pulmonary effluent, and serum melphalan were analyzed by chromatography after isolated lung perfusion or intravenous injection. On day 0, rats were injected with 2.0 x 10(6) CC531S cells intravenously. On day 7, rats underwent sham thoracotomy, received melphalan intravenously, or underwent isolated left lung perfusion with saline, melphalan, tumor necrosis factor, and a combination of the latter two. On day 14, tumor nodules were counted. RESULTS: For the doses of 400 microg tumor necrosis factor, 1,000 microg tumor necrosis factor, or both melphalan and tumor necrosis factor (2 mg + 200 microg), survival rates after contralateral pneumonectomy were 33%, 17%, and 80%, respectively. Survival in all other groups was 100%. Left lung melphalan level was significantly higher after isolated lung perfusion compared to intravenous administration. Significantly fewer left lung nodules were found for 0.5 mg isolated lung perfusion with melphalan (28+/-17) compared to isolated administration (200+/-0) (p = 0.001), and for 1.0 mg intravenous lung perfusion with melphalan (16+/-10) compared to controls (171+/-65) (p = 0.00047). Tumor necrosis factor showed no significant effect. CONCLUSIONS: Isolated lung perfusion with melphalan is an effective treatment for pulmonary metastases from adenocarcinoma in the rat. PMID- 9875779 TI - A prospective algorithm for the management of air leaks after pulmonary resection. AB - BACKGROUND: Air leaks (ALs) are a common complication after pulmonary resection, yet there is no consensus on their management. METHODS: An algorithm for the management of chest tubes (CT) and ALs was applied prospectively to 101 consecutive patients who underwent elective pulmonary resection. Air leaks were graded daily as forced expiratory only, expiratory only, inspiratory only, or continuous. All CTs were kept on 20 cm of suction until postoperative day 2 and were then converted to water seal. On postoperative day 3, if both a pneumothorax and AL were present, the CT was placed to 10 cm H2O of suction. If a pneumothorax was present without an AL, the CT was returned to 20 cm H2O of suction. Air leaks that persisted after postoperative day 7 were treated with talc slurry. RESULTS: There were 101 patients (67 men); on postoperative day 1, 26 had ALs and all were expiratory only. Univariable analysis showed a low ratio of forced expiratory volume in 1 second to forced vital capacity (FEV1/FVC) (p = 0.005), increased age (p = 0.007), increased ratio of residual volume to total lung capacity (RV/TLC) (p = 0.04), increased RV (p = 0.02), and an increased functional residual capacity (FRC) (p = 0.02) to predict the presence of an AL on postoperative day 1. By postoperative day 2, 22 patients had expiratory ALs. After 12 hours of water seal, 13 of the 22 patients' ALs had stopped, and 3 more sealed by the morning of postoperative day 3. However, 2 of the 6 patients whose ALs continued experienced a pneumothorax. Five of the 6 patients with ALs on postoperative day 4 still had ALs on postoperative day 7, and all were treated by talc slurry through the CT. All ALs resolved within 24 hours after talc slurry. CONCLUSIONS: Most ALs after pulmonary resection are expiratory only. A low FEV1/FVC ratio, increased age, increased RV/TLC ratio, increased RV, and an increased FRC were predictors of having an ALs on postoperative day 1. Conversion from suction to water seal is an effective way of sealing expiratory AL, and pneumothorax is rare. If an expiratory AL does not stop by postoperative day 4 it will probably persist until postoperative day 7, and talc slurry may be an effective treatment. PMID- 9875781 TI - Uncut Collis-Nissen fundoplication: learning curve and long-term results. AB - BACKGROUND: Between September 1985 and July 1990, the first 100 consecutive patients (50 female and 50 male) undergoing primary uncut Collis-Nissen fundoplication performed by one surgeon were reviewed. METHODS: Median age was 62 years and ranged from 19 to 89 years. Indications for repair included gastroesophageal reflux in 56 patients, obstructive symptoms in 34, and a combination of both in 10. An upper gastrointestinal endoscopy was performed in 99 patients; all were abnormal. Esophagitis was documented in 53 patients, large diaphragmatic hernia in 36, stenosis in 18, "Cameron's erosions" in 17, Barrett's disease in 13, and other findings in 9 patients. An abnormal upper gastrointestinal series was demonstrated in 96 of 97 patients evaluated. Motility studies were performed in 95 patients, and 11 had abnormal peristalsis. All procedures were performed through a left thoracotomy. RESULTS: Complications occurred in 23 patients and included respiratory failure in 6, atrial fibrillation in 3, atelectasis in 3, pneumonia in 2, myocardial infarction in 2, and chylothorax, severe dysphagia, early breakdown of repair, cardiac tamponade, hematuria, spinal headache, and intraoperative perforation by dilator in 1 each. There were 2 postoperative deaths, both cardiac in origin. Median hospitalization was 8 days (range, 6 to 76 days). The first 25 patients had 10 complications (40%) and 2 deaths (8%). The remaining 75 patients had 13 complications (17%) and no deaths (mortality, p = 0.06; morbidity, p = 0.03). Follow-up was complete in all patients for a median of 100 months (range, 3 to 138 months). Eighty-six of the 98 operative survivors are currently alive. At last follow-up, excellent functional results were observed in 58 patients (59%), good in 24 (25%), fair in 8 (8%), poor in 7 (7%), and unknown in 1 (1%). CONCLUSION: We conclude that the uncut Collis-Nissen fundoplication provides good to excellent long-term results in 84% of patients. Operative mortality and morbidity is acceptable but is associated with a learning curve. PMID- 9875780 TI - Preservation with 8-bromo-cyclic GMP improves pulmonary function after prolonged ischemia. AB - BACKGROUND: Cyclic guanosine monophosphate (cGMP) is a potent second messenger for the nitric oxide pathway in the pulmonary vasculature. Increased cytosolic cGMP levels elicit pulmonary vasodilatation resulting in decreased pulmonary vascular resistance and maximized pulmonary function after ischemia-reperfusion injury. We hypothesized that the addition of a membrane-permeable cGMP analogue (8-bromo-cGMP) to a Euro-Collins (EC) preservation solution would ameliorate pulmonary reperfusion injury better than prostaglandin E1 injection alone after prolonged hypothermic ischemia. METHODS: All lungs from New Zealand White rabbits (weight, 3 to 3.5 kg) were harvested en bloc, flushed with EC solution, and reperfused with whole blood for 30 minutes. Group 1 lungs (immediate control) were immediately reperfused. Group 2 lungs (control) were stored inflated at 4 degrees C for 18 hours before reperfusion. Groups 3 and 4 lungs were flushed with EC solution containing 200 micromol/L 8-bromo-cGMP and stored at 4 degrees C for 18 and 30 hours, respectively. Fresh, nonrecirculated venous blood was used to determine single-pass pulmonary venous-arterial oxygen gradients at 10-minute intervals. Assays for cGMP, cyclic adenosine monophosphate, nitric oxide synthase activity, and myeloperoxidase were performed on all lung tissue samples. Wet to dry weight ratios were determined after 2 weeks of passive desiccation. RESULTS: Oxygenation (venous-arterial oxygen gradient), pulmonary artery pressure, pulmonary vascular resistance, and edema formation were significantly improved in groups 3 and 4 (addition of 8-bromo-cGMP to EC plus 18 or 30 hours of hypothermic ischemia). Hypothermic storage (groups 2, 3, and 4) decreased both nitric oxide synthase activity and myeloperoxidase levels compared with immediate reperfusion (group 1). CONCLUSIONS: These results suggest that the addition of a membrane permeable cGMP analogue to an EC pulmonary flush solution improves pulmonary function after prolonged storage compared with EC and prostaglandin (E1) preservation alone. The finding of myeloperoxidase reduced levels after hypothermic storage and subsequent reperfusion may suggest a more important role for pulmonary hemodynamic control in mitigating pulmonary reperfusion injury. PMID- 9875782 TI - Lewis Y antigen expression and postoperative survival in non-small cell lung cancer. AB - BACKGROUND: In contrast to other Lewis blood group-related antigens, Lewis Y antigen (LeY) has not been fully investigated in non-small cell lung cancer. METHODS: To assess the significance of LeY expression, 236 patients with completely resected pathologic stage 1-3a were reviewed with immunohistochemical analysis. RESULTS: LeY expression was positive in 179 patients (75.8%). In poorly differentiated cancer, percentage of LeY-positive patients was lower than in moderately to well-differentiated cancer (67.2% versus 81.2%, p = 0.028). Five year survival rate of LeY-positive patients was 78.2%, significantly higher than that of LeY-negative patients (59.7%, p = 0.001). Combined with p53 status, differences in survival proved to be marked; 5-year survival rate of patients with positive LeY expression and without aberrant p53 expression, was as high as 83.3%, whereas that of patients with negative LeY expression and with aberrant p53 expression was only 38.4% (p < 0.001). Multivariate analysis confirmed that LeY expression was a significant independent factor to predict better survival. CONCLUSIONS: LeY expression is a significant prognostic factor related to grade of cancer differentiation. PMID- 9875783 TI - Early chest tube removal after video-assisted thoracoscopic wedge resection of the lung. AB - BACKGROUND: Traditional management of chest tubes after a wedge resection of peripheral pulmonary tissue often lasts several days. We evaluated the safety and efficacy of early chest tube removal in the recovery room after uncomplicated video-assisted thoracoscopic surgical wedge resections of the lung. METHODS: From December 1995 to July 1997, 59 patients underwent video-assisted thoracoscopic surgical wedge resection for indeterminate pulmonary nodules (n = 33) or interstitial lung disease (n = 26). We prospectively evaluated early chest tube removal in the last 33 patients; 18 patients with nodules and 15 with interstitial lung disease. Patients who were in the early removal group had chest tubes removed within 90 minutes of the surgical procedure. Criteria for early removal were established and met before chest tube removal. There was no difference between groups with respect to age, sex, comorbidities, or pathologic evaluation of resection specimens. RESULTS: Ninety-four percent (31 of 33) of patients considered for early chest tube removal met criteria for immediate tube removal. Air leak and excessive drainage prohibited early removal in 2 patients. Patients who were managed traditionally averaged 3.3 days with chest tubes--1.8 days on suction, 1.3 days on water seal. Patients who had early removal of their chest tubes had a shorter postoperative stay (2.0+/-1.0 versus 3.9+/-2.1 days, p = 0.001) and fewer chest roentgenograms (2.8+/-2.1 versus 5.1+/-2.0, p = 0.001). There were no differences in complications including small pneumothoraces (5 in the early removal group, 7 in the traditional management group), which were managed with observation alone. Total narcotic requirements were greater in the traditional management group (54+/-44.8 versus 24.6+/-22.9 morphine milligram equivalents, p = 0.005). CONCLUSIONS: Early chest tube removal after video assisted thoracoscopic surgical wedge resection of peripheral pulmonary tissue appears to be a safe and cost-effective practice if strict criteria for removal are met. PMID- 9875784 TI - Acute and chronic effects of bilateral lung transplantation without cardiopulmonary bypass on the first transplanted lung. AB - BACKGROUND: Bilateral lung transplantation (BLT) without cardiopulmonary bypass (CPB) may exacerbate reperfusion injury to the initially engrafted lung because of increases in pulmonary flow during implantation of the second graft. METHODS: In a retrospective review of 23 BLT patients, we hypothesized that BLT without CPB injures the first transplanted lung measured by acute and late graft dysfunction compared to the second transplanted lung. Of the 23 BLT, 19 underwent transplantation without CPB while 4 patients were placed on CPB secondary to hemodynamic instability. RESULTS: Acute graft function was assessed by radiographic scoring of lung quadrants (blinded radiologist; 0 = no infiltrate; 1 = infiltrate; maximum = 2 per lung) and by arterial/alveolar oxygen tension ratios (PaO2/ FiO2) ratios. Late graft function was evaluated by quantitative perfusion scan. Lung perfusion was graded as abnormal if less than 50% on the right or less than 45% on the left (Fisher's exact). Radiographic scores were not different between first and second implanted lungs at 1 and 24 hours, PaO2/FiO2 ratios at 1 and 24 hours were 273+/-26 and 312+/-23, respectively, and perfusion scans at 3 and 12 months revealed normal differential blood flow. CONCLUSIONS: These findings suggest no acute or chronic differences occur between the first or second transplanted lung completed without CPB. PMID- 9875785 TI - Comparison of open and thoracoscopic bilateral volume reduction surgery: complications analysis. AB - BACKGROUND: The effectiveness of lung volume reduction for the treatment of patients with emphysema is well established, but data about the surgical approach, the postoperative management, and complications are limited. We report a comparison of patients undergoing bilateral lung volume reduction (BLVRS) via median sternotomy and thoracoscopic techniques with emphasis on hospital course and complications. METHODS: All patients undergoing BLVRS at Hospital of University of Pennsylvania were analyzed for mortality and morbidity, using a combination of prospective data analysis and retrospective chart review. RESULTS: Patients undergoing BLVRS via median sternotomy were older than those undergoing video-assisted thoracoscopic surgery (VATS) procedures (63.9+/-6.89 vs 59.3+/-9.4 years, p = 0.005). Operating time was longer for the VATS procedure (147 versus 129 minutes, p = 0.006) while estimated blood less was greater for median sternotomy (209 versus 82 L, p = 0.0000017). Significant differences were found in intensive care unit stay, days intubated, life-threatening complications, respiratory complications, requirement for tracheostomy, and death that favored VATS BLVRS. When only later cohorts of patients were compared, more life threatening complications and deaths were found in patients undergoing BLVRS by median sternotomy. There were no differences between early and late median sternotomy BLVRS patients. Twenty-six percent of the lethal complications in median sternotomy BLVRS patients were bowel perforations, equally divided between duodenal ulcers and colons. CONCLUSIONS: Managing patients after BLVRS remains complex. Bilateral video-assisted volume reduction offers equivalent functional outcome with potentially decreased morbidity and mortality. Gastrointestinal perforations can complicate the management of these patients. PMID- 9875786 TI - Incidence and predictors of supraventricular dysrhythmias after pulmonary resection. AB - BACKGROUND: Patients undergoing pulmonary resection were evaluated prospectively in an effort to determine the incidence of and predictors for the development of postoperative supraventricular dysrhythmias. Specifically, we wished to test the hypothesis that the incidence of postoperative supraventricular dysrhythmias is dependent on the magnitude of pulmonary resection. METHODS: One hundred sixteen patients undergoing pulmonary resection had continuous Holter monitoring preoperatively, the day of operation, and the second postoperative day, as well as continuous cardiac monitoring throughout hospitalization. Holter interpretation was blinded to extent of resection. RESULTS: Twenty-six patients underwent pneumonectomy, 7 bilobectomy, 47 lobectomy, and 36 wedge resection. Twenty-six patients (22.4%) had supraventricular dysrhythmias, all atrial fibrillation +/- flutter. The incidence of atrial fibrillation with pneumonectomy, bilobectomy, single lobectomy, and wedge resection was 46.1%, 14.3%, 17.0%, and 13.8%, respectively (p < 0.005 pneumonectomy versus others). Overall, 31% of patients having pneumonectomy required pharmacologic therapy for dysrhythmia compared with 16% of patients having lesser resections, (p = 0.03). The peak incidence of onset of atrial fibrillation occurred on postoperative days 2 and 3 and lasted for less than 1 to 7 days, average 2.5 days. The average age of patients with dysrhythmias (64 years) was greater than those without (58 years) (p = 0.039). Thirty pre- and postoperative variables tested were not found to be significant predictors for development of postoperative atrial fibrillation. CONCLUSIONS: Atrial fibrillation occurs commonly after pulmonary resections but is not predictable. Development of atrial fibrillation is not dependent on the magnitude of pulmonary resection but is associated with the procedure pneumonectomy for reasons not elucidated. PMID- 9875787 TI - Comparison of tissue reactions in the tracheal mucosa surrounding a bioabsorbable and silicone airway stents. AB - BACKGROUND: Treatment of tracheobronchial stenosis is problematic. Conservative methods include stenting the stenotic area, but an ideal stent has not yet been developed. Bioabsorbable airway stents offer benefits; the extraction of the device is unnecessary, and the airway preserves its normal function after stent resorption. The aim of this study was to examine the suitability of self reinforced poly-L-lactide as a material for an airway stent. METHODS: A spiral airway stent made of 0.7-mm wire of self-reinforced poly-L-lactide was implanted operatively in 9 rabbits intratracheally; silicone stents served as controls. RESULTS: Silicone stents had a tendency to become stenosed with encrustation material and to develop a hyperplastic polyp at both ends. Self-reinforced poly-L lactide stents were well tolerated and caused no foreign body reaction, and they had a tendency to penetrate into the tracheal wall. They had disappeared at the end of the follow-up of 10 months. CONCLUSIONS: This experimental study showed that bioabsorbable self-reinforced poly-L-lactide is a promising material for an airway stent for treatment of airway stenosis. PMID- 9875788 TI - Experimental study of tracheal patch reconstruction with a covered expandable metallic stent. AB - BACKGROUND: We evaluated the efficacy of tracheal patch reconstruction with a covered expandable metallic stent (EMS) with omentoplasty. METHODS: After resecting the right half of the circumferential wall of two tracheal rings in adult beagle dogs, we inserted a covered EMS to reconstruct the defect interiorly. Then, through laparotomy, we made an omental pedicle flap and wrapped it around the EMS-interposed area. For comparison with the group without omentoplasty, we periodically examined the healing process macroscopically and histologically. RESULTS: Bronchofiberscopic observations revealed that incorporation of the covered EMS progressed with the passage of time and tracheal luminal patency was maintained well in both groups. However, polyplike granulation developed gradually at both ends of the EMS. Histologically, epithelium was regenerated in the patched area 4 weeks postoperatively and the area was covered with pseudostratified ciliated epithelium at 12 weeks postoperatively. Quantitative analysis of the macroscopic and histologic findings showed that the inflammatory polyps were reduced and epithelialization was promoted in the group with omentoplasty. CONCLUSIONS: Tracheal patch reconstruction with a covered EMS, when combined with omentoplasty, promoted early epithelial regeneration and suppressed the development of inflammatory polyps. PMID- 9875789 TI - Outcome of primary empyema thoracis: therapeutic and microbiologic aspects. AB - BACKGROUND: This study was undertaken to determine whether all adult patients with primary empyema thoracis need decortication. METHODS: A management algorithm was developed and analyzed in a prospective, longitudinal, nonblinded study of 179 consecutive adult patients. The treatment options included thoracentesis, closed (tube) thoracostomy, image-guided catheter drainage, and decortication. We reviewed the outcomes of these procedures as they related to the pleural fluid cultures isolated and the antibiotic regimens used. RESULTS: Of the 179 patients, 20 had thoracentesis as the primary procedure, and 18 (90%) were cured. Ninety patients underwent closed thoracostomy as the primary procedure with a cure rate of 62% (56 patients) and a mortality rate of 11% (10 patients), and 24 patients required a secondary procedure. Seventy-six patients underwent decortication as either the primary or the secondary procedure with a cure rate of 88% (67 patients) and a mortality rate of 1.3% (1 patient); 8 patients required conversion to open thoracostomy. Hospital stay for decortication was 14+/-1 days and for closed thoracostomy, 17+/-1 days (p < 0.05). Decortication was necessary in 55% of patients with anaerobic infections and in 50% with aerobic infections. Clindamycin in combination with gentamicin sulfate was the most efficacious regimen with a success rate of 82% (51 of 62 patients); only 33% (17 of 52) were cured with penicillin. The overall mortality rate in this study was 6.7% (12 of 179 patients). CONCLUSIONS: Forty-two percent of patients with primary empyema thoracis ultimately require decortication. Decortication is more frequently necessary for anaerobic, tuberculous, staphylococcal, and pneumococcal infections. Although the overall mortality in this study was low, mortality remains high in elderly patients and patients with comorbid disease. PMID- 9875790 TI - Ongoing prospective study of segmentectomy for small lung tumors. Study Group of Extended Segmentectomy for Small Lung Tumor. AB - BACKGROUND: Lesser resection for small lung tumors remains an unresolved problem. This study was conducted to see whether this type of operation is acceptable or not. METHODS: From 1992 to 1994, 55 patients were enrolled in a multicenter trial of limited surgical resection for peripheral tumors of less than 2 cm diameter. The procedure consisted of segmentectomy with exploration of lymph nodes by examining frozen sections. The operation was modified if the report was positive. The intersegmental plane was identified by keeping the resected segments inflated and the preserved segments collapsed. To divide the plane, stapling or electrocauterization on the edge of the collapsed area was used. In this way the resection line was delivered beyond the burdened segment; this was called extended segmentectomy. RESULTS: There were no perioperative deaths, but there were eight postoperative deaths. In 1 patient who died because of local recurrence, it had been known that the margin to the lesion had been narrow (15 mm); 1 had bilateral intrapulmonary nodules, 1 had nodules in the side that was not operated on, and another succumbed to a second neoplasm of small cell lung cancer 4 years after the first operation. The remaining 4 died of nonpulmonary diseases. Almost all other patients are alive and free from recurrence, except for 1 in whom N2 disease was not detected intraoperatively but was confirmed after the operation. CONCLUSIONS: The interim results suggest that extended segmentectomy is applicable in patients with a small peripheral lung cancer. However, a wide margin and aggressive intraoperative pathologic examinations are mandatory. PMID- 9875791 TI - Long-term remission after resection, chemotherapy, and irradiation of a metastatic myxoma. AB - Malignant local or metastatic myxomatous tumors of the heart are rare and sometimes present with an unpredictable outcome. The present report demonstrates an unusual case of left atrial myxoma with a rather strange distribution of metastases that had long-term remission after combined surgical resection, chemotherapy, and irradiation of cerebral and pulmonary metastases. PMID- 9875792 TI - Aortico-right ventricular tunnel. AB - A successful operation on an infant with a tunnel through which the aorta communicated with the right ventricle is reported. The diagnosis was suspected preoperatively on the basis of two-dimensional Doppler color echocardiography and confirmed by cardiac catheterization. The aortico-right ventricular tunnel originated independently from the left coronary ostium and above the sinus of Valsalva. Patch closure from inside the tunnel under deep hypothermia was successfully performed. Follow-up is satisfactory 5 years later. PMID- 9875793 TI - Murphy's law in cardiac surgery. AB - We report the case of a 41-year-old man who underwent heart transplantation after suffering from aortic dissection during the implantation of a biventricular assist device for heart failure after coronary artery bypass grafting. The biventricular assist device had to be operated by hand for 10 hours because of a technical defect. In the end perseverance won out and the patient is now doing well at 2.5 years of follow-up. PMID- 9875794 TI - Anomalous right coronary artery from the pulmonary artery in Taussig-Bing anomaly. AB - The presence of associated anomalies in patients with double-outlet right ventricle can significantly alter surgical intervention. Preoperative delineation of these anomalies can facilitate surgical planning and improve outcome. We describe a case in which the right coronary artery and anterior descending coronary artery arose from the pulmonary artery in a patient with double-outlet right ventricle with subpulmonary ventricular septal defect (Taussig-Bing anomaly). Recognition of this important anomaly prevented significant intraoperative myocardial damage by altering techniques of cardioplegia administration for myocardial preservation. PMID- 9875795 TI - Anatomy of inferior pulmonary vein should be clarified in lower lobectomy. AB - We successfully performed surgery for carcinoma of the lung on 2 patients with the anatomic variation of the middle lobe vein draining to the right inferior pulmonary vein. This variation is surgically important because division of the right inferior pulmonary vein may result in blockage of middle lobe vein drainage in right lower lobectomy. Surgeons must always pay attention to this variation when performing the right lower lobectomy. PMID- 9875796 TI - Successful palliation of acute superior vena caval obstruction after the Senning operation. AB - Superior vena caval obstruction after an intraatrial baffle procedure can lead to acute cerebral complications and hence requires immediate management. We present a case of successful palliation of acute superior vena caval obstruction after a Senning procedure by establishing a venous shunt between the innominate vein and pulmonary artery. This technique resulted in immediate hemodynamic and functional improvement that subsequently allowed for the enlargement of the superior vena cava-right atrial junction with a pericardial patch. PMID- 9875797 TI - Hepatoatrial anastomosis and tricuspid valve replacement in Budd-Chiari syndrome 8. AB - The rare complication of tricuspid valve stenosis due to displacement of a peritoneovenous shunt catheter developed in a patient with Budd-Chiari syndrome. Surgical management included shunt removal and tricuspid valve replacement combined with hepatoatrial anastomosis. The tricuspid valve was replaced with a bioprosthesis, and liver resection opened the stenotic confluence of the hepatic veins. The right atrium was sutured to the liver capsule, thereby restoring physiologic hepatic venous blood flow. PMID- 9875798 TI - Chest wall repair with a titanium instrument. AB - We performed chest wall repair with titanium alloy instruments as artificial ribs for prevention of paradoxical respiration and protection of the lung and liver after chest wall resection including the nearly entire length of the right seventh to the eleventh ribs and the costal arch for metastasis of osteosarcoma. The technique of this operation is presented diagrammatically. PMID- 9875799 TI - Snaring of a coronary artery causing distal atheroma embolization. AB - We report a case of total occlusion of the left anterior descending coronary artery with atheromatous debris after "off-pump" grafting. Atheroma break-off and distal embolization probably occurred from proximal snare occlusion of the target coronary artery, and this was recognized by intraoperative angiography. Alternatives to suture snaring should be considered whenever an atherosclerotic artery is to be grafted without cardiopulmonary bypass. PMID- 9875800 TI - Mitral valve repair after excision of a fibrolipoma. AB - A 2-year-old patient with severe mitral regurgitation associated with a mass originating from the anterior leaflet of the mitral valve is reported. Excision of the tumor and mitral valve repair was performed. Four years later the child remains asymptomatic with no recurrence of the tumor or regurgitation. PMID- 9875801 TI - Six-year survival after excision of cardiac malignant lymphoma. AB - We report a patient with a postoperative survival period of 6 years after the surgical excision of a cardiac malignant lymphoma. A 35-year-old woman underwent total excision of the tumor arising from the left ventricular outflow tract. After the operation, she was treated with chemotherapy for 6 months. She has been doing well thereafter without any medication. To date there is no evidence of recurrence. PMID- 9875802 TI - Traumatic tricuspid valve regurgitation and cerebral emboli. AB - We report a case of traumatic tricuspid valve regurgitation and late presentation with transient ischemic attacks caused by the presence of a right-to-left shunt through a small atrial septal defect. PMID- 9875803 TI - Blunt injury rupture of tricuspid valve and right coronary artery. AB - Successful surgical repair of an unusual case of blunt trauma to the chest and abdomen is presented. The injury resulted in rupture of the pericardium, avulsion and rupture of the right coronary artery into the right atrium, complete disruption of the tricuspid valve, and acute right heart failure with complete heart block. PMID- 9875804 TI - ABIOMED BVS 5000 system: repair of venous cannulation site for excessive bleeding. AB - The most common morbidity of the ABIOMED (Danvers, MA) left ventricular assist device is severe hemorrhage. Severe bleeding around the left atrial cannulation site can be repaired easily with the proper technique. PMID- 9875805 TI - Homograft failure in mycotic aortic aneurysm caused by Streptococcus pneumoniae. AB - Treatment of a rare Streptococcus pneumoniae mycotic aneurysm by homograft replacement failed in a 59-year-old patient because of persistent lobar pneumonia. Despite reoperation with replacement of the infected homograft by a fresh one, he finally died of septicemia. This case illustrates that homograft tissue may be infected per continuum and that extensive debridement of periaortic tissue-including major lung resection-and the use of muscle flaps may be necessary in certain circumstances. PMID- 9875806 TI - Delayed diagnosis of anomalous origin of the left coronary artery 16 years after mitral valve replacement. AB - Mitral insufficiency caused by ischemia is frequently found in anomalous origin of the left coronary artery from the pulmonary artery. We report a case of a 25 year-old woman who was diagnosed to have anomalous origin of the left coronary artery from the pulmonary artery and had successful left internal mammary artery bypass grafting 16 years after mitral valve replacement for mitral insufficiency of an unknown cause in her childhood. PMID- 9875807 TI - Improved exposure of the pulmonary arteries for thromboendarterectomy. AB - Pulmonary thromboendarterectomy is a surgical technique for treating pulmonary hypertension caused by unresolved pulmonary embolism. It has been recommended to perform this procedure under deep hypothermic circulatory arrest. Here we describe two technical modifications: (1) improved exposure to the right pulmonary artery by division of the superior caval vein and (2) thromboendarterectomy in normothermic cardiopulmonary bypass, with beating heart or electrically induced ventricular fibrillation. These modifications allow complete endarterectomy of both pulmonary arteries under normothermic conditions, thus avoiding hypothermic circulatory arrest, which results in short cardiopulmonary bypass times and reduces the morbidity and mortality of this procedure. PMID- 9875808 TI - Elective drainage of the apical chest by posterior approach. AB - Pneumothorax due to incomplete reexpansion of remaining lung after a thoracic operation is difficult to drain via standard access routes. Apical chest drainage can be placed through a upper posterior point of the back, located in the laterovertebral area at the level of the first or second intercostal space. This technique, which is very safe and well tolerated, avoids the necessity to use thoracoplasty or muscle flap plombage of those residual cavities. PMID- 9875809 TI - Extracardiac conduit Fontan procedure without cardiopulmonary bypass. AB - There are a number of potential advantages of extracardiac conduit cavopulmonary anastomosis for palliation of functional single ventricle heart disease, including the ability to perform the operation with no aortic cross-clamping and with minimal duration of extracorporeal circulation. In many patients, it may be possible to perform the procedure without cardiopulmonary bypass altogether. In this report, we present our technique for performing the extracardiac conduit Fontan operation without cardiopulmonary bypass. PMID- 9875810 TI - Alternative technique of right-sided outflow cannula insertion for right ventricular support. AB - Right ventricular assist devices are an important part of the armamentarium of cardiac surgeons for the treatment of right-sided circulatory failure after cardiac transplantation or insertion of a left ventricular assist device. However, right ventricular assist device insertion can be technically challenging in the setting of pulmonary hypertension because of a number of concomitant anatomic and physiologic phenomena. We present a technique for the insertion of the right ventricular assist device outflow cannula that is easier and faster to insert, and safer to explant, especially if cardiopulmonary bypass is to be avoided. PMID- 9875811 TI - Pericardial patch augmentation of anterior tricuspid leaflet in Ebstein's anomaly. AB - Although refinement of tricuspid valvuloplasty and plication techniques has opened the way to a satisfactory outlook for the majority of patients with Ebstein's anomaly, paucity of tissue in the anterior tricuspid leaflet may preclude successful valve repair. In this particular circumstance, a modified Carpentier repair of the tricuspid valve with additional pericardial patch augmentation of the anterior leaflet may result in a well-functioning monocusp tricuspid valve. PMID- 9875812 TI - Thoracoscopic pleural tent. AB - Lung volume reduction has been performed in patients with advanced emphysema to relieve dyspnea and improve exercise tolerance. Median sternotomy and video assisted thoracoscopy have been proposed as equally adequate approaches; however, prolonged postoperative air leakage is the most prevalent complication in all series. For this reason, on the basis of the experience achieved with the median sternotomy approach, buttressing of the suture line with different materials and techniques for space reduction have been proposed. We describe a technique to create a pleural tent after thoracoscopic volume reduction. The thoracoscopic creation of a pleural tent is feasible and results in a duration of postoperative air leaks and hospital stays similar to that achieved with stapler line buttressing. PMID- 9875813 TI - Speed-controlled venovenous modified ultrafiltration for pediatric open heart operations. AB - Although modified ultrafiltration after pediatric open heart operations is used by several clinical centers, the risk of complications is a matter of concern. This report describes a simple, reliable, and reproducible technique of speed controlled venovenous modified ultrafiltration. PMID- 9875814 TI - Measures to control blood activation during assisted circulation. AB - Major improvements in heart assist devices have allowed prolonged mechanical circulatory support with successful subsequent weaning or heart transplantation. The contact of blood with biomaterials used in life-sustaining devices and numerous biomaterial-independent factors elicit a systemic inflammatory response, which involves activation of various plasma protein systems and blood cells. Prolonged mechanical circulatory support elicits a systemic inflammatory response and hemostatic perturbations similar to that reported during cardiopulmonary bypass. However, in the setting of prolonged assistance, time has a complex and ill-known influence on blood activation. Methods to reduce blood activation during prolonged assisted circulation are derived from cardiopulmonary bypass investigations. Improving the biocompatibility of artificial devices can be achieved either by biomaterial surface modifications, by inhibition of biologic cascades leading to blood activation, or by controlling end points of biologic cascades. However, the necessity to respect the integrity of the organism during prolonged assistance precludes most systemic interventions and limits the control of blood activation to the area of the device. PMID- 9875815 TI - Chylothorax. PMID- 9875816 TI - As originally published in 1991: Use of the inferior epigastric artery as a free graft for myocardial revascularization. 1998 update. PMID- 9875817 TI - Policy statement: minimally invasive coronary artery bypass surgery. The Society of Thoracic Surgeons/American Association for Thoracic Surgery Committee on New Technology. PMID- 9875818 TI - Course guidelines for minimally invasive cardiac surgery. The Society of Thoracic Surgeons/American Association for Thoracic Surgery Committee on New Technology. PMID- 9875819 TI - Long-term mechanical circulatory support system reliability recommendation: American Society for Artificial Internal Organs and The Society of Thoracic Surgeons: long-term mechanical circulatory support system reliability recommendation. AB - Jointly developed by members of the American Society for Artificial Internal Organs and the Society of Thoracic Surgeons along with staff from the Food and Drug Administration, the National Heart, Lung and Blood Institute and other experts, this recommendation describes the reliability considerations and goals for Investigational Device Exemption and Premarket Approval submissions for long term, mechanical circulatory support systems. The recommendation includes a definition of system failure, a discussion of an appropriate reliability model, a suggested in vitro reliability test plan, reliability considerations for animal implantation tests, in vitro and animal in vivo performance goals, the qualification of design changes during the Investigational Device Exemption clinical trial, the development of a Failure Modes Effects and Criticality Analysis, and the reliability information for surgeons and patient candidates. The document will be periodically reviewed to assess its timeliness and appropriateness within five years. PMID- 9875820 TI - Interlocking sternal closure. PMID- 9875821 TI - Fiftieth anniversary of successful operation for mitral stenosis. PMID- 9875822 TI - Cerebral effects of superior vena caval cannula obstruction. PMID- 9875823 TI - Removing the causes of pulmonary hypertension. PMID- 9875824 TI - Tetrazolium reductase activity in human heart valves. PMID- 9875825 TI - Cardiopulmonary bypass for tracheal tumor resection: revival of an old technique? PMID- 9875826 TI - Prevention of potential complications after minimal access aortic valve replacement. PMID- 9875827 TI - Innovative uses of saphenous vein grafts in minimally invasive coronary surgery. PMID- 9875828 TI - Reducing inflammatory reactions by heparin-coated circuit. PMID- 9875829 TI - Atrial flap anastomosis: an alternative technique for orthotopic heart transplantation. PMID- 9875830 TI - Is resection the only effective treatment for bronchial carcinoids? PMID- 9875831 TI - Refined alpha aminooleic acid and experimental calcification in bioprostheses. PMID- 9875832 TI - Angiogenesis induced by transmyocardial laser revascularization. PMID- 9875833 TI - Clinical trials and treatment effects monitoring. AB - Treatment effects monitoring is the act of reviewing accumulated data by treatment group to determine whether a trial should continue unaltered. That monitoring is required for any trial where there is a risk of an aggregate form of harm for subjects because of continued use of an inferior treatment or because of failure to use a superior treatment. Institutional review boards (IRBs), in deciding whether to approve such research, require that (when appropriate) there is adequate provision for monitoring the data collected to ensure the safety of subjects. The focus here is on the conditions necessary to satisfy this requirement. The conditions discussed are timeliness of the monitoring, completeness of data for monitoring, competency of monitors, and freedom of monitors to act and recommend as they deem necessary, regardless of the wishes, desires, or dictates of sponsors. PMID- 9875834 TI - Successful techniques for retention of study participants in an inner-city population. AB - The purpose of this work was to describe methods of retaining participants in studies of inner-city populations, including the timing and intensity of contacts; and to describe the characteristics of participants who did not complete all follow-up interviews and/or return all peak flow diaries in the National Cooperative Inner-City Asthma Study. A cohort study design was used involving hospital emergency rooms and community clinics in seven major urban areas. Participants included 1337 4- to 9-year-old asthmatic children and their caretakers. Nearly 89% of participants completed 3-, 6-, and 9-month follow-up interviews. The 15% of participants who completed a baseline interview on the weekends were significantly more likely to complete follow-up interviews on a weekend. The percent of follow-up interviews conducted in person increased over time from 5% to 8%. The percent of participants with complete follow-up increased as the number of contact names increased (86% with zero contacts, 91% with two contracts; p = 0.03, test for trend). Participants who required at least four phone calls to complete the 3- and 6-month assessment were significantly more likely to be black, have higher participant stress, and have a smoker in the household (p < 0.05). Multiple logistic regression suggests that higher social support and lower parental stress were both predictors of completed interviews. Within our study sample of inner-city minority participants with asthmatic children, only a small proportion of participants missed any follow-up interviews. Increased caretaker stress, decreased social support, and inability to provide several alternate contacts were all predictive of retention problems. Having a flexible staff, computer tracking, and face-to-face recruitment appear essential to achieving nearly complete follow-up within a population historically difficult to follow. PMID- 9875835 TI - An evaluation of a measure of the proportion of the treatment effect explained by a surrogate marker. AB - Time-dependent markers, such as CD4 and viral load, are potential surrogate markers in AIDS clinical trials. A critical issue with surrogate markers is whether changes in these markers explain the beneficial effect of treatment on the real end point of the clinical trial. A statistic to measure the proportion of the treatment effect explained by the surrogate is p(FGS) = 1 - gamma/alpha, where alpha is the treatment effect coefficient in a Cox model and gamma is the treatment effect coefficient from a time-dependent Cox model adjusted for the marker. In this article we evaluate the statistical properties of p(FGS). Using a Monte Carlo study we show that the statistic is not well calibrated, because it can fall outside the range zero to one, even in very large samples. In the simulation study we consider situations where the time-dependent marker is measured with error at a fixed number of times. We show that a method of fitting a time-dependent Cox model involving smoothing the marker reduces the bias in the estimate of p(FGS) compared with the standard method of using the current or last observed marker value. We also show that the estimate of p(FGS) has considerable variability and can have wide confidence intervals. We conclude that p(FGS) is only likely to be useful in large trials with a strong treatment effect. The methods are illustrated using CD4 counts from an AIDS clinical trial of zidovidine versus placebo. PMID- 9875836 TI - Sensitivity, specificity, and vaccine efficacy. AB - We study the effects of imperfect sensitivity and specificity on estimation of vaccine efficacy (VE). We show that a specificity of <1.0 (i.e., some noncases are classified as cases) can reduce the value of the estimate of VE. One may attempt to adjust the estimate by accounting for the values of sensitivity and specificity. This increases the variance of the estimate of VE and leads to larger sample size requirements. Alternatively, one may adopt the diagnosis method as the disease definition. PMID- 9875837 TI - Early stopping of a clinical trial when there is evidence of no treatment benefit: protocol B-14 of the National Surgical Adjuvant Breast and Bowel Project. AB - Although several randomized clinical trials in the 1980s indicated a benefit from the use of tamoxifen in the treatment of early-stage breast cancer, questions have remained regarding the optimal duration of drug administration. In 1982, the National Surgical Adjuvant Breast and Bowel Project (NSABP) initiated a randomized trial to compare 5 years of tamoxifen to placebo among breast cancer patients with estrogen receptor-positive tumors and no evidence of axillary node involvement. By 1987, evidence of a substantial benefit for tamoxifen led the NSABP to extend this trial to determine whether longer duration tamoxifen therapy would be additionally beneficial. This study randomized patients who had completed 5 years of tamoxifen free of breast cancer recurrence or other events to either tamoxifen or placebo for an additional 5 years. By 1994, 1172 women had entered the study and accrual was closed. In late 1995, the trial was terminated on the basis of interim findings indicating that a benefit for continuing tamoxifen would not be realized. The closure has prompted controversy among cancer researchers, because there are currently at least three tamoxifen duration trials in progress, whereas results from two other studies evaluating 5-year duration therapy versus longer therapy were recently published. Here, we provide details of the statistical rationale contributing to our decision to recommend early closure of the study. We then consider other possible approaches to assessing the appropriateness of early termination in the face of evidence against a benefit, including Bayesian methods, which can be used to incorporate a range of prior beliefs regarding the efficacy of a treatment with accruing information from the trial. We also briefly discuss results of the other published studies. PMID- 9875838 TI - Power and sample size calculations for studies involving linear regression. AB - This article presents methods for sample size and power calculations for studies involving linear regression. These approaches are applicable to clinical trials designed to detect a regression slope of a given magnitude or to studies that test whether the slopes or intercepts of two independent regression lines differ by a given amount. The investigator may either specify the values of the independent (x) variable(s) of the regression line(s) or determine them observationally when the study is performed. In the latter case, the investigator must estimate the standard deviation(s) of the independent variable(s). This study gives examples using this method for both experimental and observational study designs. Cohen's method of power calculations for multiple linear regression models is also discussed and contrasted with the methods of this study. We have posted a computer program to perform these and other sample size calculations on the Internet (see http://www.mc.vanderbilt.edu/prevmed/psintro+ ++.htm). This program can determine the sample size needed to detect a specified alternative hypothesis with the required power, the power with which a specific alternative hypothesis can be detected with a given sample size, or the specific alternative hypotheses that can be detected with a given power and sample size. Context-specific help messages available on request make the use of this software largely self-explanatory. PMID- 9875840 TI - Coiled coils in both intracellular vesicle and viral membrane fusion. PMID- 9875841 TI - Immune receptor editing: revise and select. AB - Secondary antigen receptor gene recombination occurs in developing lymphocytes, and in more mature T and B cells. In the developing lymphocyte, "editing" occurs in response to receptor ligation by autoantigens. As a result of receptor editing, anti-self-reactive cells are converted to non-self-reactive cells, and self-reactive clones are thereby salvaged before negative selection. This antigen induced change in the receptor specificity was not foreseen in the clonal selection theory. However, editing could be incorporated into the clonal selection theory by limiting receptor selection to a specific stage in lymphocyte development that precedes cellular selection as suggested by Jerne (1971). We know much less about the origin, regulation, or function of V(D)J recombination in mature lymphocytes. Nevertheless, antigen-induced receptor selection is likely to play an important role in shaping immune responses. PMID- 9875839 TI - The Women's Health Initiative Memory Study (WHIMS): a trial of the effect of estrogen therapy in preventing and slowing the progression of dementia. AB - Evidence from animal, human cross-sectional, case-control, and prospective studies indicate that hormone replacement therapy (HRT) is a promising treatment to delay the onset of symptoms of dementia. The Women's Health Initiative Memory Study (WHIMS) is the first double-masked, randomized, placebo-controlled, long term clinical trial designed to test the hypothesis that HRT reduces the incidence of all-cause dementia in women aged 65 and older. WHIMS, an ancillary study to the Women's Health Initiative (WHI) funded by the National Institutes of Health, will recruit a subgroup of women aged 65 and older from among those enrolling in the HRT trial of the WHI. The WHI clinical centers and 10 affiliated satellites plan to enroll approximately 8300 women into WHIMS over a 2-year period. Participants will be followed annually for 6 years, receiving cognitive assessments via the Modified Mini-Mental State (3MS) Examination. Women who screen positively for cognitive impairment on the basis of an educational and age adjusted 3MS cutpoint proceed to more extensive neuropsychological testing and neurologic evaluation. Each woman suspected to have dementia then undergoes a series of laboratory tests that confirm the clinical diagnosis and classify the type of dementia. WHIMS is designed to provide more than 80% statistical power to detect a 40% reduction in the rate of all-cause dementia, an effect that could have profound public health implications for older women's health and functioning. PMID- 9875842 TI - Using genetic mutations to study the neural basis of behavior. PMID- 9875843 TI - The curious status of the Golgi apparatus. PMID- 9875844 TI - A critical role for DNA end-joining proteins in both lymphogenesis and neurogenesis. AB - XRCC4 was identified via a complementation cloning method that employed an ionizing radiation (IR)-sensitive hamster cell line. By gene-targeted mutation, we show that XRCC4 deficiency in primary murine cells causes growth defects, premature senescence, IR sensitivity, and inability to support V(D)J recombination. In mice, XRCC4 deficiency causes late embryonic lethality accompanied by defective lymphogenesis and defective neurogenesis manifested by extensive apoptotic death of newly generated postmitotic neuronal cells. We find similar neuronal developmental defects in embryos that lack DNA ligase IV, an XRCC4-associated protein. Our findings demonstrate that differentiating lymphocytes and neurons strictly require the XRCC4 and DNA ligase IV end-joining proteins and point to the general stage of neuronal development in which these proteins are necessary. PMID- 9875845 TI - Plexin A is a neuronal semaphorin receptor that controls axon guidance. AB - The Semaphorins comprise a large family of secreted and transmembrane proteins, some of which function as repellents during axon guidance. Semaphorins fall into seven subclasses. Neuropilins are neuronal receptors for class III Semaphorins. In the immune system, VESPR, a member of the Plexin family, is a receptor for a viral-encoded Semaphorin. Here, we identify two Drosophila Plexins, both of which are expressed in the developing nervous system. We present evidence that Plexin A is a neuronal receptor for class I Semaphorins (Sema 1a and Sema 1b) and show that Plexin A controls motor and CNS axon guidance. Plexins, which themselves contain complete Semaphorin domains, may be both the ancestors of classical Semaphorins and binding partners for Semaphorins. PMID- 9875846 TI - Identification of ligands for olfactory receptors by functional expression of a receptor library. AB - The recognition of odorants by olfactory receptors represents the first stage in odor discrimination. Here, we report the generation of an expression library containing a large and diverse repertoire of mouse olfactory receptor sequences in the transmembrane II-VII region. From this library, 80 chimeric receptors were tested against 26 odorants after transfection into HEK-293 cells. Three receptors were identified to respond to micromolecular concentrations of carvone, (-) citronellal, and limonene, respectively. We also found that the mouse I7 receptor, unlike the rat I7 receptor, prefers heptanal instead of octanal, as a result of a single valine-to-isoleucine substitution. This finding represents the beginning of a molecular understanding of odorant recognition. The identification, on a large scale, of cognate receptor-odorant interactions should provide insight into olfactory coding mechanisms. PMID- 9875847 TI - The structural basis of estrogen receptor/coactivator recognition and the antagonism of this interaction by tamoxifen. AB - Ligand-dependent activation of transcription by nuclear receptors (NRs) is mediated by interactions with coactivators. Receptor agonists promote coactivator binding, and antagonists block coactivator binding. Here we report the crystal structure of the human estrogen receptor alpha (hER alpha) ligand-binding domain (LBD) bound to both the agonist diethylstilbestrol (DES) and a peptide derived from the NR box II region of the coactivator GRIP1 and the crystal structure of the hER alpha LBD bound to the selective antagonist 4-hydroxytamoxifen (OHT). In the DES-LBD-peptide complex, the peptide binds as a short alpha helix to a hydrophobic groove on the surface of the LBD. In the OHT-LBD complex, helix 12 occludes the coactivator recognition groove by mimicking the interactions of the NR box peptide with the LBD. These structures reveal the two distinct mechanisms by which structural features of OHT promote this "autoinhibitory" helix 12 conformation. PMID- 9875848 TI - Crystal structure of constitutive endothelial nitric oxide synthase: a paradigm for pterin function involving a novel metal center. AB - Nitric oxide, a key signaling molecule, is produced by a family of enzymes collectively called nitric oxide synthases (NOS). Here, we report the crystal structure of the heme domain of endothelial NOS in tetrahydrobiopterin (H4B)-free and -bound forms at 1.95 A and 1.9 A resolution, respectively. In both structures a zinc ion is tetrahedrally coordinated to pairs of symmetry-related cysteine residues at the dimer interface. The phylogenetically conserved Cys-(X)4-Cys motif and its strategic location establish a structural role for the metal center in maintaining the integrity of the H4B-binding site. The unexpected recognition of the substrate, L-arginine, at the H4B site indicates that this site is poised to stabilize a positively charged pterin ring and suggests a model involving a cationic pterin radical in the catalytic cycle. PMID- 9875849 TI - Crystal structure of the human high-affinity IgE receptor. AB - Allergic responses result from the activation of mast cells by the human high affinity IgE receptor. IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. The structure reveals a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. A prominent loop that confers specificity for IgE molecules extends from the receptor surface near an unusual arrangement of four exposed tryptophans. The crystal structure of the IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. PMID- 9875850 TI - Crystal structure of the Oxytricha nova telomere end binding protein complexed with single strand DNA. AB - Telomeres are specialized protein-DNA complexes that compose the ends of eukaryotic chromosomes. Telomeres protect chromosome termini from degradation and recombination and act together with telomerase to ensure complete genome replication. We have determined the crystal structure of the two-subunit Oxytricha nova telomere end binding protein (OnTEBP) complexed with single strand telomeric DNA at 2.8 A resolution. The structure reveals four oligonucleotide/oligosaccharide-binding folds, three of which form a deep cleft that binds the ssDNA, and a fourth that forms an unusual protein-protein interaction between the alpha and beta subunits. This structure provides a molecular description of how the two subunits of OnTEBP recognize and bind ssDNA to form a sequence-specific, telomeric nucleoprotein complex that caps the very 3' ends of chromosomes. PMID- 9875851 TI - Reconstitution of an SOS response pathway: derepression of transcription in response to DNA breaks. AB - E. coli responds to DNA damage by derepressing the transcription of about 20 genes that make up the SOS pathway. Genetic analyses have shown that SOS induction in response to double-stranded DNA (dsDNA) breaks requires LexA repressor, and the RecA and RecBCD enzymes--proteins best known for their role as initiators of dsDNA break repair and homologous recombination. Here we demonstrate that purified RecA protein, RecBCD enzyme, single-stranded DNA binding (SSB) protein, and LexA repressor respond to dsDNA breaks in vitro by derepressing transcription from an SOS promoter. Interestingly, derepression is more rapid if the DNA containing the dsDNA break has a chi recombination hot spot (5'-GCTGGTGG-3'), suggesting a novel regulatory role for one of the most overrepresented octamers in the E. coli genome. PMID- 9875852 TI - lin-35 and lin-53, two genes that antagonize a C. elegans Ras pathway, encode proteins similar to Rb and its binding protein RbAp48. AB - The Ras signaling pathway for vulval induction in Caenorhabditis elegans is antagonized by the activity of the synthetic multivulva (synMuv) genes, which define two functionally redundant pathways. We have characterized two genes in one of these pathways. lin-35 encodes a protein similar to the tumor suppressor Rb and the closely related proteins p107 and p130. lin-53 encodes a protein similar to RbAp48, a mammalian protein that binds Rb. In mammals, Rb and related proteins act as regulators of E2F transcription factors, and RbAp48 may act with such proteins as a transcriptional corepressor. We propose that LIN-35 and LIN-53 antagonize the Ras signaling pathway in C. elegans by repressing transcription in the vulval precursor cells of genes required for the expression of vulval cell fates. PMID- 9875853 TI - Procollagen traverses the Golgi stack without leaving the lumen of cisternae: evidence for cisternal maturation. AB - Newly synthesized procollagen type I (PC) assembles into 300 nm rigid, rod-like triple helices in the lumen of the endoplasmic reticulum. This oligomeric complex moves to the Golgi and forms large electron-dense aggregates. We have monitored the transport of PC along the secretory pathway. We show that PC moves across the Golgi stacks without ever leaving the lumen of the Golgi cisternae. During transport from the endoplasmic reticulum to the Golgi, PC is found within tubular saccular structures greater than 300 nm in length. Thus, supermolecular cargoes such as PC do not utilize the conventional vesicle-mediated transport to traverse the Golgi stacks. Our results imply that PC moves in the anterograde direction across the Golgi complex by a process involving progressive maturation of Golgi cisternae. PMID- 9875854 TI - Evidence for periciliary liquid layer depletion, not abnormal ion composition, in the pathogenesis of cystic fibrosis airways disease. AB - The pathogenesis of cystic fibrosis (CF) airways infection is unknown. Two hypotheses, "hypotonic [low salt]/defensin" and "isotonic volume transport/mucus clearance," attempt to link defects in cystic fibrosis transmembrane conductance regulator-mediated ion transport to CF airways disease. We tested these hypotheses with planar and cylindrical culture models and found no evidence that the liquids lining airway surfaces were hypotonic or that salt concentrations differed between CF and normal cultures. In contrast, CF airway epithelia exhibited abnormally high rates of airway surface liquid absorption, which depleted the periciliary liquid layer and abolished mucus transport. The failure to clear thickened mucus from airway surfaces likely initiates CF airways infection. These data indicate that therapy for CF lung disease should not be directed at modulation of ionic composition, but rather at restoring volume (salt and water) on airway surfaces. PMID- 9875855 TI - Use of dsRNA-mediated genetic interference to demonstrate that frizzled and frizzled 2 act in the wingless pathway. AB - We investigated the potential of double-stranded RNA to interfere with the function of genes in Drosophila. Injection of dsRNA into embryos resulted in potent and specific interference of several genes that were tested. In contrast, single-stranded RNA weakly interfered with gene activity. The method was used to determine the reception mechanism of the morphogen Wingless. Interference of the frizzled and Drosophila frizzled 2 genes together produced defects in embryonic patterning that mimic loss of wingless function. Interference of either gene alone had no effect on patterning. Epistasis analysis indicates that frizzled and Drosophila frizzled 2 act downstream of wingless and upstream of zeste-white3 in the Wingless pathway. Our results demonstrate that dsRNA interference can be used to analyze many aspects of gene function. PMID- 9875856 TI - frizzled and frizzled 2 play a partially redundant role in wingless signaling and have similar requirements to wingless in neurogenesis. AB - The Drosophila Frizzled (Fz) and Frizzled2 (DFz2) proteins function as receptors for Wingless (Wg) in tissue culture cells. While previous results indicate that loss of function for fz has tissue polarity defects, loss-of-function effects of Dfz2 are not known. Here, we have examined the requirements of fz and Dfz2 during neurogenesis. Our results indicate that both Fz and DFz2 function in Wg signaling, and loss of either of the two affects the same subset of neuroblasts as those affected by loss of wg. While these defects are partially penetrant in embryos lacking either fz or Dfz2, the penetrance is significantly enhanced in embryos lacking both. Since the penetrance of the CNS phenotypes is not complete in double mutants, additional components that allow some degree of Wg signaling must exist in vivo. PMID- 9875857 TI - A quarter century of health maintenance. PMID- 9875858 TI - Erecting an ethical framework for managed care. PMID- 9875860 TI - Extending Brady background checks opposed. PMID- 9875859 TI - NHLBI at 50: reflections on a half-century of research on the heart, lungs, and blood. National Heart, Lung, and Blood Institute. Interview by Charles Marwick. PMID- 9875861 TI - Air Force school celebrates "supporting the flier". PMID- 9875862 TI - Antileukotriene drugs in the management of asthma. PMID- 9875863 TI - A piece of my mind. Four-wheel-drive medicine. PMID- 9875864 TI - Improving the accuracy of abstracts in scientific articles. PMID- 9875865 TI - Benefits and limitations of prenatal care. PMID- 9875866 TI - Benefits and limitations of prenatal care. PMID- 9875867 TI - Benefits and limitations of prenatal care. PMID- 9875868 TI - Contempo 1998: new treatments for epilepsy. PMID- 9875869 TI - Methylprednisolone for unresolving ARDS. PMID- 9875870 TI - Racial and ethnic differences in serum cotinine levels. PMID- 9875871 TI - Racial and ethnic differences in serum cotinine levels. PMID- 9875872 TI - Racial and ethnic differences in serum cotinine levels. PMID- 9875873 TI - Bad vibrations after bypass surgery. PMID- 9875874 TI - Effect of alendronate on risk of fracture in women with low bone density but without vertebral fractures: results from the Fracture Intervention Trial. AB - CONTEXT: Alendronate sodium reduces fracture risk in postmenopausal women who have vertebral fractures, but its effects on fracture risk have not been studied for women without vertebral fractures. OBJECTIVE: To test the hypothesis that 4 years of alendronate would decrease the risk of clinical and vertebral fractures in women who have low bone mineral density (BMD) but no vertebral fractures. DESIGN: Randomized, blinded, placebo-controlled trial. SETTING: Eleven community based clinical research centers. SUBJECTS: Women aged 54 to 81 years with a femoral neck BMD of 0.68 g/cm2 or less (Hologic Inc, Waltham, Mass) but no vertebral fracture; 4432 were randomized to alendronate or placebo and 4272 (96%) completed outcome measurements at the final visit (an average of 4.2 years later). INTERVENTION: All participants reporting calcium intakes of 1000 mg/d or less received a supplement containing 500 mg of calcium and 250 IU of cholecalciferol. Subjects were randomly assigned to either placebo or 5 mg/d of alendronate sodium for 2 years followed by 10 mg/d for the remainder of the trial. MAIN OUTCOME MEASURES: Clinical fractures confirmed by x-ray reports, new vertebral deformities detected by morphometric measurements on radiographs, and BMD measured by dual x-ray absorptiometry. RESULTS: Alendronate increased BMD at all sites studied (P<.001) and reduced clinical fractures from 312 in the placebo group to 272 in the intervention group, but not significantly so (14% reduction; relative hazard [RH], 0.86; 95% confidence interval [CI], 0.73-1.01). Alendronate reduced clinical fractures by 36% in women with baseline osteoporosis at the femoral neck (>2.5 SDs below the normal young adult mean; RH, 0.64; 95% CI, 0.50 0.82; treatment-control difference, 6.5%; number needed to treat [NNT], 15), but there was no significant reduction among those with higher BMD (RH, 1.08; 95% CI, 0.87-1.35). Alendronate decreased the risk of radiographic vertebral fractures by 44% overall (relative risk, 0.56; 95% CI, 0.39-0.80; treatment-control difference, 1.7%; NNT, 60). Alendronate did not increase the risk of gastrointestinal or other adverse effects. CONCLUSIONS: In women with low BMD but without vertebral fractures, 4 years of alendronate safely increased BMD and decreased the risk of first vertebral deformity. Alendronate significantly reduced the risk of clinical fractures among women with osteoporosis but not among women with higher BMD. PMID- 9875875 TI - Prior misdemeanor convictions as a risk factor for later violent and firearm related criminal activity among authorized purchasers of handguns. AB - CONTEXT: Under current federal law, many persons with prior convictions for misdemeanor offenses pass criminal records background checks and legally purchase handguns. OBJECTIVE: To determine whether authorized handgun purchasers with prior misdemeanor convictions are more likely than those with no criminal history to be charged with new crimes, particularly offenses involving firearms and violence. DESIGN: Retrospective cohort study. SETTING AND PARTICIPANTS: A total of 5923 authorized purchasers of handguns in California in 1977 who were younger than 50 years, identified by random sample. MAIN OUTCOME MEASURES: Incidence and relative risk (RR) of first charges for new criminal offenses after handgun purchase. RESULTS: Of the 5923 authorized purchasers, 3128 had at least 1 conviction for a misdemeanor offense prior to handgun purchase, and 2795 had no prior criminal history. Follow-up to the end of the 15-year observation period or to death was available for 77.8% of study subjects and for a median 8.9 years for another 9.6%. Handgun purchasers with at least 1 prior misdemeanor conviction were more than 7 times as likely as those with no prior criminal history to be charged with a new offense after handgun purchase (RR, 7.5; 95% confidence interval [CI], 6.6-8.7). Among men, those with 2 or more prior convictions for misdemeanor violence were at greatest risk for nonviolent firearm-related offenses such as weapon carrying (RR, 11.7; 95% CI, 6.8-20.0), violent offenses generally (RR, 10.4; 95% CI, 6.9-15.8), and Violent Crime Index offenses (murder or non-negligent manslaughter, forcible rape, robbery, or aggravated assault) (RR, 15.1; 95% CI, 9.4-24.3). However, even handgun purchasers with only 1 prior misdemeanor conviction and no convictions for offenses involving firearms or violence were nearly 5 times as likely as those with no prior criminal history to be charged with new offenses involving firearms or violence. CONCLUSIONS: Handgun purchasers with prior misdemeanor convictions are at increased risk for future criminal activity, including violent and firearm-related crimes. PMID- 9875876 TI - Pretreatment evaluation of chronic hepatitis C: risks, benefits, and costs. AB - CONTEXT: Chronic hepatitis C (CHC) infection affects nearly 4 million people in the United States. Treatment with interferon alfa-2b has been limited by its cost and low likelihood of long-term response. OBJECTIVE: To examine the cost effectiveness of alternative pretreatment management strategies for patients with CHC. DESIGN: Decision and cost-effectiveness analysis using a Markov model to examine prevalence of genotypes, viral load, and histological characteristics in relation to the sustained response rate with treatment. Data were based on a previously published decision model and a MEDLINE literature search for hepatitis C, biopsy, and liver from 1966 to 1996. PATIENTS: A hypothetical population of patients with CHC infection and elevated serum alanine aminotransferase level. INTERVENTIONS: Combinations of liver biopsy, genotyping, and quantitative viral load determination prior to a single 6-month course of interferon alfa-2b; empirical interferon treatment; and conservative management. MAIN OUTCOME MEASURES: Proportion of sustained responders, lifetime costs, life expectancy, and quality-adjusted life expectancy. RESULTS: Strategies involving hepatitis C virus (HCV) RNA testing had marginal cost-effectiveness ratios up to $4400 per discounted quality-adjusted life-year gained but would miss up to 36% of sustained responders. Empirical interferon treatment had a marginal cost effectiveness ratio of $12400 per discounted quality-adjusted life-year gained and reached all potential sustained responders. Strategies involving liver biopsy were more expensive and would miss 6% of sustained responders and yield slightly lower life expectancies. CONCLUSIONS: Routine liver biopsy before treatment with interferon increases the cost of managing patients with CHC without improving health outcomes. Using quantitative HCV RNA testing to guide therapy misses some potential sustained responders. Empirical interferon treatment has a marginal cost-effectiveness ratio within the bounds of other commonly accepted therapies and misses none of the sustained responders. PMID- 9875877 TI - Serogroup Y meningococcal disease in Chicago, 1991-1997. AB - CONTEXT: In 1994, surveillance by the Chicago Department of Public Health detected a growing trend in the proportion of invasive meningococcal infections caused by serogroup Y. OBJECTIVE: To examine the emergence of serogroup Y meningococcal disease and compare its clinical characteristics with those of other meningococcal serogroups. DESIGN: Population-based retrospective review of surveillance records; medical record review and cohort analysis of serogroup Y vs non-serogroup Y case patients. SETTING: Chicago, III. PARTICIPANTS: City residents with Neisseria meningitidis isolated from a normally sterile site from January 1, 1991, through December 31, 1997; cohort analysis included those identified through March 31, 1996. MAIN OUTCOME MEASURES: Serogroup-specific incidence, demographics, and clinical outcomes. RESULTS: We identified 214 case patients; 53 (25%) had serogroup Y. The attack rate of serogroup Y meningococcal disease increased from 0.04 cases per 100000 in 1991 to a peak of 0.82 cases per 100000 in 1995 and subsequently decreased to 0.26 cases per 100000 and 0.34 cases per 100000 in 1996 and 1997, respectively. Compared with patients infected by other serogroups, patients with serogroup Y were older (median age, 16 years vs 1 year; P = .001) and more likely to have a chronic underlying illness (prevalence ratio, 2.3; 95% confidence interval, 1.2-4.4). Outcome did not differ significantly between the 2 groups. Multilocus enzyme electrophoresis typing of isolates from 19 case patients identified 5 different types. We found no clustering among the enzyme types by age, race/ethnicity, community area, or time. CONCLUSIONS: Serogroup Y emerged as the most frequent cause of meningococcal disease in Chicago in 1995 and accounted for a substantial proportion of cases in 1996 and 1997. Current data suggest that the magnitude of serogroup Y meningococcal disease is sufficient for vaccine developers to incorporate serogroup Y into new vaccines. PMID- 9875878 TI - The National Cholesterol Education Program: progress and prospects. AB - The National Cholesterol Education Program (NCEP) is a prime example of the role the National Heart, Lung, and Blood Institute has played, in its 50 years of existence, as a catalyst for translating research advances into improved clinical and public health practices. Since its inception in 1985, the NCEP has adhered to 2 principles in mounting educational campaigns for professionals and the public: building on a strong science base and working in partnership with other organizations. In slightly more than a decade, the NCEP has made significant progress toward its goal of reducing the prevalence of high blood cholesterol. The impact of cholesterol education is clearly visible in 4 major trends: increasing professional and public cholesterol awareness; declining dietary intakes of saturated fat, total fat, and cholesterol; falling serum cholesterol levels; and a continuing decline in coronary heart disease (CHD) mortality rates. Nevertheless, cholesterol levels are still being undertreated, especially in patients with CHD, and substantial scientific and educational challenges remain. As it looks forward to the 21st century, the NCEP plans to make continued progress by using emerging scientific developments and pursuing the powerful combination of cholesterol lowering in CHD patients and in primary prevention. PMID- 9875879 TI - Effect of epidural vs parenteral opioid analgesia on the progress of labor: a meta-analysis. AB - CONTEXT: Epidural labor analgesia, if selected by the patient, is associated with high cesarean delivery rates. Results of randomized trials comparing rates of cesarean delivery using epidural anesthesia vs parenteral opioids are inconsistent. OBJECTIVE: To review the effects of epidural vs parenteral opioid analgesia on cesarean delivery rates. DATA SOURCES: Studies were identified by searching MEDLINE from January 1966 through January 1998, the Cochrane Database of Perinatal Trials, and relevant nonindexed journals and abstracts. STUDY SELECTION: We included all studies that randomized patients to epidural vs parenteral opioid labor analgesia. DATA EXTRACTION: Two authors independently extracted data from 10 trials enrolling 2369 patients. Odds ratios (ORs) for categorical data, weighted mean differences (WMDs) for continuous data, and 95% confidence intervals (CIs) were calculated using a random-effects model. DATA SYNTHESIS: The risk of cesarean delivery did not differ between patients receiving epidural (8.2%) vs parenteral opioid (5.6%) analgesia (OR, 1.5; 95% CI, 0.81-2.76). Epidural patients had longer first (WMD, 42 minutes; 95% CI, 17-68 minutes) and second (WMD, 14 minutes; 95% CI, 5-23 minutes) labor stages. While epidural patients were more likely to have instrumented delivery (OR, 2.19; 95% CI, 1.32-7.78), they were no more likely to have instrumented delivery for dystocia (OR, 0.68; 95% CI, 0.31-1.49). After epidural analgesia, neonates were less likely to have low 5-minute Apgar scores (OR, 0.38; 95% CI, 0.18-0.81) or to need naloxone (OR, 0.24; 95% CI, 0.07-0.77). Women receiving epidural analgesia had lower pain scores during the first (WMD, -40 mm on a 100-mm scale; 95% CI, 42 to -38 mm) and second (WMD, -29 mm; 95% CI, -38 to -21 mm) stages of labor. The odds of dissatisfaction were lower with epidural analgesia (OR, 0.25; 95% CI, 0.20-0.32). CONCLUSIONS: Epidural labor analgesia is not associated with increased rates of instrumented vaginal delivery for dystocia or cesarean delivery. Patients receiving epidural analgesia have longer labors. Patient satisfaction and neonatal outcome are better after epidural than parenteral opioid analgesia. PMID- 9875880 TI - A 69-year-old man with chronic dizziness. PMID- 9875881 TI - A 66-year-old woman with ulcerative colitis, 1 year later. PMID- 9875882 TI - Bone mass, bone fragility, and the decision to treat. PMID- 9875883 TI - Handgun purchasers with misdemeanor convictions. PMID- 9875884 TI - Violence, a neglected epidemic: call for papers. PMID- 9875886 TI - JAMA patient page. Hepatitis C. PMID- 9875885 TI - Americans' views on children's health. PMID- 9875887 TI - The sirens' songs of confirmatory sepsis trials: selection bias and sampling error. PMID- 9875888 TI - Hyperbaric oxygen in systemic inflammation ... HBO is not just a movie channel anymore. PMID- 9875889 TI - Prone position in acute respiratory distress syndrome: turning over an old idea. PMID- 9875890 TI - Cardiac troponin I and non-Q-wave myocardial infarction: how useful is it after coronary artery bypass surgery? PMID- 9875891 TI - Gastric tonometry in the fed or fasting state? PMID- 9875892 TI - Hemofiltration for SIRS: bloodletting, twentieth century style? PMID- 9875893 TI - Hospital wards spreading vancomycin-resistant enterococci to intensive care units: returning coals to Newcastle. PMID- 9875894 TI - Hypothermia and extracorporeal rewarming: the journey toward a less invasive, more accessible methodology. PMID- 9875895 TI - Nitric oxide synthase inhibition in septic shock--a question of the right time, the right drug, and the right model? PMID- 9875896 TI - Is transient acidosis detrimental to the nervous system? PMID- 9875897 TI - Should we measure ionized magnesium? PMID- 9875898 TI - To monitor or not to monitor. PMID- 9875899 TI - Noninvasive management of pediatric neuromuscular ventilatory failure: a viable alternative. PMID- 9875900 TI - In search of the Holy Grail: noninvasive or "low-invasive" cardiac output monitoring. PMID- 9875901 TI - High-frequency ventilation. PMID- 9875902 TI - Reduced mortality from septic shock--lessons for the future. PMID- 9875903 TI - Propofol toxicity in critically ill pediatric patients: show us the proof. PMID- 9875904 TI - In memoriam: Gunther Schlag, MD. PMID- 9875905 TI - Confirmatory platelet-activating factor receptor antagonist trial in patients with severe gram-negative bacterial sepsis: a phase III, randomized, double blind, placebo-controlled, multicenter trial. BN 52021 Sepsis Investigator Group. AB - OBJECTIVE: To determine the efficacy and safety of using natural platelet activating factor receptor antagonist (PAFra), BN 52021, to treat patients with severe Gram-negative bacterial sepsis. DESIGN: A prospective, randomized, double blind, placebo-controlled, multicenter clinical trial. SETTING: Fifty-nine academic medical center intensive care units in Europe. PATIENTS: Six hundred nine patients with severe sepsis, suspected to be related to Gram-negative bacterial infection, who received PAFra or placebo. INTERVENTIONS: Patients were randomized to receive either a dose of PAFra (120 mg iv) every 12 hrs over a 4 day period or placebo over a 4-day period. MEASUREMENTS AND MAIN RESULTS: The patients were well matched at study entry for severity of illness and for risk factors known to influence the outcome of sepsis. Among all randomized patients, the 28-day, all-cause mortality rate was 49% (152/308) in the placebo group, and 47% (140/300) in the PAFra group (p=.50). When analyzed on the basis of the previously defined target population, the 28-day, all-cause mortality rate was 50% (115/232) in the placebo group and 44% (94/212) in the PAFra group, yielding a 12% reduction in mortality rate (p=.29). In patients with documented infection involving other organisms, there was no difference between treated and placebo groups. When the outcomes of organ dysfunctions were examined in the overall population and in the documented Gram-negative bacterial infection population, the number of patients who resolved hepatic dysfunction tended to be higher in the treated group than in the placebo group (p=.06). The number of adverse events reported were not different between the two groups. CONCLUSIONS: A 4-day administration of the studied PAFra (BN 52021) failed to demonstrate a statistically significant reduction in the mortality rate of patients with severe sepsis suspected to be related to Gram-negative bacterial infection. If PAFra treatment has any therapeutic activity in severe Gram-negative bacterial sepsis, the incremental benefits are small and will be difficult to demonstrate in a patient population as defined by this clinical trial. PMID- 9875906 TI - Effects of hyperbaric oxygen exposure on a zymosan-induced shock model. AB - OBJECTIVE: To evaluate the effects of hyperbaric oxygen (HBO) therapy on zymosan induced shock in rats. Zymosan, a cell wall component of the yeast Saccharomyces cerevisiae, induces inflammation by causing the production of various cytokines and pro-inflammatory mediators. The administration of zymosan to rats represents a new experimental shock model by inducing acute peritonitis, severe hypotension, and signs of systemic illness. However, it has been recently proposed that the zymosan-induced shock, like septic shock, may be mediated by overproduction of nitric oxide. DESIGN: Experimental study. SETTING: Institute of Pharmacology and Toxicology, 2nd University of Naples, Naples, Italy. SUBJECTS: Male rats were treated with zymosan (500 mg/kg) by intraperitoneal route, with HBO (2 Absolute Atmosphere) or with zymosan and HBO (2 Absolute Atmosphere). MEASUREMENTS AND MAIN RESULTS: Peritoneal exudate, plasma, and peritoneal nitric oxide metabolites (NOx) and zymosan determined a time-dependent increase in peritoneal and plasma NOx concentrations, and peritoneal leukocytes were determined. Moreover, symptomatology was observed. The administration of zymosan caused the appearance of a severe illness in the rats characterized by ruffled fur, lethargy, conjunctivitis, diarrhea, and a significant loss of body weight. All zymosan treated rats developed an acute peritonitis, producing turbid exudate. Zymosan determined a time-dependent increase in peritoneal, plasma NOx, and tumor necrosis factor (TNF)-alpha concentrations. Morbidity of zymosan shocked rats has been attenuated and no mortality was observed by treatment with HBO. These findings were associated with a significant reduction either of peritoneal leukocytes and exudate, or plasma and peritoneal NOx concentrations. Moreover, TNF-alpha levels were significantly reduced in animals shocked by zymosan and treated with HBO. PMID- 9875907 TI - Effects of the prone position on gas exchange and hemodynamics in severe acute respiratory distress syndrome. AB - OBJECTIVES: To address the following issues regarding the use of prone position ventilation in patients with severe acute respiratory distress syndrome (ARDS): a) response rate; b) magnitude and duration of improved oxygenation in responders during a 12-hr trial and the consequences of returning to the supine position; c) effects of the prone position on gas exchange and hemodynamics; d) consequences of oxygenation in nonresponders; and e) effects of repeated prone position trials. DESIGN: Prospective, nonrandomized interventional study. SETTING: Medical intensive care unit, university tertiary care center. PATIENTS: Nineteen consecutive, mechanically ventilated patients (age 45+/-20 yrs, mean+/-SD) with ARDS and severe hypoxemia, defined as PaO2/FiO2 of < or = 150 with FiO2 of > or = 0.6 persisting for < or =24 hrs, and a pulmonary artery occlusion pressure of <18 mm Hg. INTERVENTIONS: Patients were turned prone for 2 hrs. Nonresponders were returned supine, but responders were maintained prone for 12 hrs before being returned to the supine position. The procedure was repeated on a daily basis in all patients, until inclusion criteria were no longer met or the patients died. MEASUREMENTS AND MAIN RESULTS: Hemodynamic, blood gas, and gas exchange measurements were performed at the following time points: a) baseline supine; b) after 30 mins prone; and c) after 120 mins prone. Additional measurements for nonresponders were taken after 30 mins supine. For responders, additional measurements were taken after 12 hrs prone and 30 mins supine. Patients were considered responders if an increase in PaO2 of > or = 10 torr (> or =1.3 kPa), or increase in the PaO2/FiO2 ratio of >20 occurred within 120 mins. Eleven (57%) patients responded to the prone position. There was no difference in initial baseline parameters between responders and nonresponders. After 30 mins, the prone position in responders increased PaO2 and decreased calculated venous admixture (Qva/Qt). This improvement was the maximal obtained, and was maintained throughout the 12-hr prone period. After 12 hrs prone, mean FiO2 had been lowered from 0.85+/-0.16 to 0.66+/-0.18 (p < .05). Thirty minutes after the patients were returned supine, PaO2, PaO2/FiO2, and Qva/Qt were not different from 12-hr prone values, and were improved in comparison with baseline supine values. There was no worsening of gas exchange or hemodynamics in nonresponders. After the initial trial, a total of 28 additional episodes of prone position ventilation were performed in nine of the 19 patients. Of the 24 additional episodes in the responders, there was a response in 17 (71%) of 24 episodes. In the four additional episodes in nonresponders, there was a response in only one (25%) of four episodes. Response was accompanied by the same beneficial effects on gas exchange and Qva/Qt and absence of effect on hemodynamics as in the initial trial. There was no worsening in gas exchange or hemodynamics in nonresponder trials. CONCLUSIONS: Based on the data from this study, the prone position can improve oxygenation in severely hypoxemic ARDS patients without deleterious effects on hemodynamics. This beneficial effect does not immediately disappear on return to the supine position. In our patients, an absence of response to this technique was not accompanied by worsening hypoxemia or hemodynamic instability. Repeated daily trials in the prone position should be considered in the management of ARDS patients with severe hypoxemia. PMID- 9875908 TI - Cardiac troponin I and Q-wave perioperative myocardial infarction after coronary artery bypass surgery. AB - OBJECTIVE: To monitor cardiac troponin I (cTnI), a newly developed biochemical index for cardiac damage, in patients during and after coronary artery bypass surgery (CABS) to determine whether the measurement of the serum levels of this marker could be of value in formulating an early diagnosis of Q-wave perioperative myocardial infarction (PMI). DESIGN: Prospective study with sequential measurements of biological markers in a selected surgical patient group. SETTING: University research laboratory and general university hospital (Cardiac Surgery Unit and Anesthesiology and Reanimation Unit). PATIENTS: Forty two patients undergoing elective CABS without concomitant valvular replacement. INTERVENTIONS: There were no interventions required for this study. However, patients entered into the study had CABS, sequential arterial blood samples, ECG recordings, and echocardiograms performed. MEASUREMENTS AND MAIN RESULTS: Pre-, intra-, and postoperative (up to 48 hrs) measurements of cardiac troponin I, MB CK, and total creatine kinase, as well as serial electrocardiograms and echocardiograms. Perioperative infarction was assessed as the development of new persistent regional wall motion abnormalities in echocardiography together with electrocardiographic alterations and MB-CK increases. Eight patients had Q-wave PMI. All PMI patients had elevated peak cTnI values (all >9.2 ng/mL), whereas the 34 nonPMI patients had peak values <9.0 ng/mL; therefore, sensitivity and specificity (with a 9.0 ng/mL cut-off value) are 100%. MB-CK measurement peak values did not demonstrate such a high specificity and sensitivity. CONCLUSIONS: Because of its high specificity and sensitivity, serial measurements of cTnI provide a rapid and accurate method for confirming or excluding the diagnosis of perioperative myocardial injury. cTnI evaluation can therefore be used both as an independent prognostic marker for patients undergoing cardiac surgery and as a powerful tool for detecting smaller PMIs often missed with standard PMI diagnostic criteria. PMID- 9875909 TI - Gastric versus duodenal feeding and gastric tonometric measurements. AB - OBJECTIVE: To compare the influence of gastric and postpyloric enteral feeding on the gastric tonometric PCO2 gap (tonometric PCO2 - PaCO2). DESIGN: A prospective, clinical trial. SETTING: Two intensive care units in a university hospital. PATIENTS: Twenty patients undergoing mechanical ventilation and enteral feeding without catecholamines, sepsis, or sign of hypoxia. INTERVENTIONS: Patients were randomized to receive feeding through the tonometer (gastric group), or through a postpyloric tube (postpyloric group). MEASUREMENTS AND MAIN RESULTS: The patients received tube feeding at a rate of 50 mL/hr during 4 hrs. Baseline measurements included: mean arterial pressure, heart rate, tonometric parameters, arterial gases, and arterial lactate concentration. Except for lactate concentration, these measurements were repeated after 1 and 4 hrs of enteral feeding and 2 hrs after stopping enteral feeding. During the study, arterial pH and PaCO2 did not change. During enteral feeding, the PCO2 gap increased in the gastric group from a mean of 7+/-5 to 17+/-14 (SD) torr (0.9 0.7 to 2.3+/-1.9 kPa) (p< .O01) and did not change in the postpyloric group (5+/-5 to 3+/-1 torr [0.7+/-0.7 to 0.4+/-0.1 kPa]). Two hours after stopping enteral feeding, the PCO2 gap was still increased in the gastric group (15+/-9 vs. 7+/-5 torr [2.0+/-1.2 vs. 0.9+/-0.7 kPa]) (p < .01). CONCLUSION: The results indicate that gastric enteral feeding increased the PCO2 gap. However, postpyloric enteral feeding does not interact with gastric tonometric measurements and should be used when using gastric tonometry in enterally fed patients. PMID- 9875910 TI - Diffusive vs. convective therapy: effects on mediators of inflammation in patient with severe systemic inflammatory response syndrome. AB - OBJECTIVE: To compare two forms of continuous renal replacement therapy, continuous venovenous hemofiltration (CVVH) vs. continuous venovenous hemodialysis (CVVHD), in terms of the removal of inflammatory mediators from the blood of patients with systemic inflammatory response syndrome and acute renal failure. DESIGN: Randomized crossover, clinical study. SETTING: University teaching hospital. PATIENTS: Thirteen patients with systemic inflammatory response syndrome and acute renal failure receiving continuous renal replacement therapy. INTERVENTION: Patients were randomized to receive either convective clearance using CVVH or diffusive clearance using CVVHD for the first 24 hrs, followed by the other modality for 24 hrs. All treatments utilized AN69 hemofilters. CVVH was performed with an ultrafiltration rate of 2 L/hr and CVVHD with a dialysis outflow rate of 2 L/hr. MEASUREMENTS AND MAIN RESULTS: Plasma and ultrafiltrate concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-10, and sL-selectin were measured at 0, 1, 3, 6, 12, and 24 hrs by radioimmunoassay. Plasma endotoxin concentrations were also measured at 0, 12, and 24 hrs by chromogenic assay. CVVH was associated with a 13% decrease in plasma TNF-alpha concentrations compared with a 23% increase while on CVVHD (p < .05). Mean plasma concentrations of IL-6, IL-10, and sL-selectin were unchanged over time and between therapies. Only minimal amounts of mediators were recovered in the effluents with either therapy except for IL-6. The clearances for IL-6 were different between therapies, 1.9+/-0.8 (SD) mL/min for CVVHD and 3.3+/-1.5 mL/min for CVVH, (p< .01). Plasma endotoxin concentrations were not different between therapies. CONCLUSION: CVVH resulted in a decrease in plasma TNF-alpha concentrations as compared with CVVHD, while the type of transport mechanism used did not influence plasma concentrations of IL-6, IL-10, soluble L-selectin, or endotoxin. Differences in clearance for IL-6 between CVVH and CVVHD did not translate into significant changes in circulating IL-6 concentrations. PMID- 9875911 TI - External sources of vancomycin-resistant enterococci for intensive care units. AB - OBJECTIVE: The incidence of colonization and infection with vancomycin-resistant enterococci (VRE) has increased dramatically in the last 5 yrs, especially in intensive care units (ICUs). We studied VRE-colonization in patients on admission to a medical ICU (MICU) where VRE colonization is endemic. DESIGN: Prospective, descriptive analysis. SETTING: An MICU of a public hospital. PATIENTS: Three hundred and one consecutively admitted patients. MEASUREMENTS AND MAIN RESULTS: Rectal swabs were obtained on admission from all patients. VRE isolates from all colonized patients were genetically fingerprinted by pulsed-field gel electrophoresis (PFGE). Forty-three (14%) of 301 patients were colonized with VRE on MICU admission. Three (7%) of these 43 patients were admitted directly from the community without prior hospital contact. Risk of colonization on admission was related to the length of stay in the hospital before MICU-admission (odds ratio 4.65 for patients with a stay of at least 3 days) and previous in-hospital use of antibiotics. Of 22 VRE PFGE strain types recognized in the MICU during the study period, four (18%) were introduced by patients admitted directly from the community and ten (45%) were introduced by patients admitted from other hospital wards. CONCLUSIONS: These results show that although ICUs are considered epicenters for antibiotic resistance, sources extraneous to our MICU (e.g., other wards) contributed the majority of VRE strain types in the unit. PMID- 9875912 TI - Activation of the extrinsic coagulation pathway in patients with severe sepsis and septic shock. AB - OBJECTIVES: To obtain systematic information on the extrinsic coagulation pathway, as well as to investigate the time course of the coagulation abnormalities in sepsis. DESIGN: Prospective observational study. SETTING: General intensive care unit. PATIENTS: Nineteen patients with the diagnosis of severe sepsis or septic shock and nine control patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Tissue factor antigen concentration (tissue factor antigen), prothrombin fragment F1+2, thrombin antithrombin III complex, fibrinopeptide A, D-dimer, and antithrombin III concentrations were measured on the day of diagnosis of severe sepsis and septic shock, and on days 1, 2, 3, and 4 after diagnosis. The concentrations of tissue factor antigen, prothrombin fragment F1+2, fibrinopeptide A, and D-dimer were significantly increased in patients with severe sepsis and septic shock compared with control subjects. However, the concentrations of thrombin antithrombin III complex showed no statistical differences between the septic patients and the control subjects. Significantly, low antithrombin III concentrations were observed in the septic patient groups compared with control subjects. With the exception of D-dimer, the concentrations of the hemostatic markers were similar between severe sepsis and septic shock patients. Significant correlations were noted between tissue factor antigen and the disseminated intravascular coagulation score (r2=.236, p< .0001) and the number of dysfunctioning organs (r2=.229, p=.035). CONCLUSIONS: We systematically elucidated coagulation disorders in newly defined sepsis. The extrinsic coagulation pathway is activated in patients with severe sepsis and septic shock. In these patients, enhanced thrombin generation and activation, and fibrin formation were demonstrated when compared with the control subjects. Furthermore, the thrombin generated appears not to be fully neutralized by antithrombin III. PMID- 9875913 TI - Intra-abdominal Candida infection during acute necrotizing pancreatitis has a high prevalence and is associated with increased mortality. AB - OBJECTIVES: To analyze the frequency of intra-abdominal Candida infection during acute necrotizing pancreatitis and to compare the outcome to patients without Candida infections. DESIGN: Retrospective analysis of data from 37 patients with and without Candida infection. SETTING: Surgical intensive care unit (ICU) in a university hospital. PATIENTS: Thirty-seven patients with acute necrotizing pancreatitis. INTERVENTIONS: Patients were subject to necrosectomy and programmed lavage. MEASUREMENTS AND MAIN RESULTS: Clinical parameters contained in the Acute Physiology and Chronic Health Evaluation (APACHE) II score were monitored, as were microbiological results from the intra-abdominal primary focus and from sites of secondary infections. Body mass index, distribution of underlying diseases, length of ICU stay, number of operations, and outcome were recorded. Multivariate logistic regression analysis for mortality as the dependent variable was performed. Intra-abdominal Candida infection was observed in 13 of 37 cases and was associated with a four-fold greater mortality rate compared with intra abdominal bacterial infection alone. Multivariate logistic regression analysis demonstrated that Candida infection contributed to mortality significantly (p < .025) and independently from APACHE II (p < .006; mortality odds ratio for the two parameters=12.5). Lack of antimycotic treatment was associated with an increase to 29.2 (p<.001) in the odds ratio. CONCLUSIONS: Given the impact of Candida infection on mortality to acute necrotizing pancreatitis and the apparent benefit from antimycotic chemotherapy, the data argue for an early fungicide chemotherapeutic intervention. PMID- 9875914 TI - Continuous venovenous rewarming: results from a juvenile animal model. AB - OBJECTIVE: To compare a standard and an experimental method of rewarming in 5-wk old goats with induced moderate hypothermia. DESIGN: Hypothermia was induced in ten juvenile Nubian goats. Five goats were randomly assigned to be rewarmed using standard techniques, and five were assigned to the experimental rewarming technique of a modified continuous venovenous hemofiltration circuit. SETTING: Animal research facility, Greenville Hospital System/Clemson University Biomedical Cooperative, Clemson, S.C. SUBJECTS: Ten 5-wk-old goats. INTERVENTIONS: Hypothermia to a body temperature of 29.4 degrees C was induced in the goats. Each of the control group of five goats was rewarmed using standard methods. Each of the experimental group of five goats was rewarmed using a modified continuous venovenous hemofiltration circuit. MEASUREMENTS AND MAIN RESULTS: At 2 hrs, the median temperature increase in the experimental group was 6.5 degrees C, compared with an increase of only 1.5 degrees C in the control group (p=.02). The mean increase in core body temperature over time (from baseline to 150 mins) was also significantly greater in the experimental group (p=.006). CONCLUSIONS: The use of a modified continuous venovenous hemofiltration circuit for rewarming in a juvenile goat model after induction of moderate hypothermia is more effective than are standard methods. PMID- 9875915 TI - Treatment of septic shock in rats with nitric oxide synthase inhibitors and inhaled nitric oxide. AB - OBJECTIVE: To evaluate the effect of treatment with a combination of nitric oxide synthase inhibitors and inhaled nitric oxide on systemic hypotension during sepsis. DESIGN: Prospective, randomized, controlled study on anesthetized animals. SETTING: A cardiopulmonary research laboratory. SUBJECTS: Forty-seven male adult Sprague-Dawley rats. INTERVENTIONS: Animals were anesthetized, mechanically ventilated with room air, and randomized into six groups: a) the control group (C, n=6) received normal saline infusion; b) the endotoxin-treated group received 100 mg/kg i.v. of Escherichia coli lipopolysaccharide (LPS, n=9); c) the third group received LPS, and 1 hr later the animals were treated with 100 mg/kg i.v. Nw-nitro-L-arginine (LNA, n=9); d) the fourth group received LPS, and after 1 hr, the animals were treated with 100 mg/kg i.v. aminoguanidine (AG, n=9); e) the fifth group received LPS and 1 hr later was treated with LNA plus 1 ppm inhaled nitric oxide (LNA+NO, n=7); f) the sixth group received LPS and 1 hr later was treated with aminoguanidine plus inhaled NO (AG+NO, n=7). Inhaled NO was administered continuously until the end of the experiment. MEASUREMENTS AND MAIN RESULTS: Systemic mean blood pressure (MAP) was monitored through a catheter in the carotid artery. Mean exhaled NO (ENO) was measured before LPS (T0) and every 30 mins thereafter for 5 hrs. Arterial blood gases and pH were measured every 30 mins for the first 2 hrs and then every hour. No attempt was made to regulate the animal body temperature. All the rats became equally hypothermic (28.9+/-1.2 degrees C [SEM]) at the end of the experiment. In the control group, blood pressure and pH remained stable for the duration of the experiment, however, ENO increased gradually from 1.3+/-0.7 to 17.6+/-3.1 ppb after 5 hrs (p< .05). In the LPS treated rats, MAP decreased in the first 30 mins and then remained stable for 5 hrs. The decrease in MAP was associated with a gradual increase in ENO, which was significant after 180 mins (58.9+/-16.6 ppb) and reached 95.3+/-27.5 ppb after 5 hrs (p< .05). LNA and AG prevented the increase in ENO after LPS to the level in the control group. AG caused a partial reversal of systemic hypotension, which lasted for the duration of the experiment. LNA reversed systemic hypotension almost completely but only transiently for 1 hr, and caused severe metabolic acidosis in all animals. The co-administration of NO with AG had no added benefits on MAP and pH. In contrast, NO inhalation increased the duration of the reversal in MAP after LNA, alleviated the degree of acidosis, and decreased the mortality rate (from 55% to 29%). CONCLUSIONS: In this animal model, LPS-induced hypotension was alleviated slightly and durably after AG, but only transiently after LNA. Furthermore, co-administration of NO with AG had no added benefits but alleviated the severity of metabolic acidosis and mortality after LNA. We conclude that nitric oxide synthase (NOS) inhibitors, given as a single large bolus in the early phase of sepsis, can exhibit some beneficial effects. Administration of inhaled NO with NOS inhibitors provided more benefits in some conditions and therefore may be a useful therapeutic combination in sepsis. NO production in sepsis does not seem to be a primary cause of systemic hypotension. Other factors are likely to have a major role. PMID- 9875916 TI - Basic fibroblast growth factor reduces lactic acid-induced neuronal injury in rat hippocampal neurons. AB - OBJECTIVE: To evaluate the long-term effects of lactic acidosis and to examine a potential neuroprotective role of basic fibroblast growth factor (bFGF) on hippocampal neurons. DESIGN: Long-term observation in a cell-culture study. SETTING: University research laboratory. SUBJECTS: Adult, differentiated, primary rat hippocampal neurons. INTERVENTIONS: Neurons were exposed to medium acidified with 20 mM lactic acid, pH 6.2, for a 10-min period, and maintained untreated or in the presence of bFGF (500 pg/mL, 1 ng/mL, 10 ng/mL, 20 ng/mL) applied after exposure. MEASUREMENTS AND MAIN RESULTS: Viability was analyzed by a dye inclusion/enzyme activity test and morphology by phase contrast and immunofluorescence microscopy. [3H]Arachidonic acid (AA) release was measured by liquid scintillation spectrometry. All cultures appeared to be unchanged during the first days after exposure to lactic acidosis. Neurodegeneration became apparent within 3 days. Seven days after exposure, cell survival decreased to 60% in lactic acidosis-injured, untreated cultures. Morphologic damage appeared as a 50% reduction in axonal and 25% reduction in dendritic arborizations. AA release increased to four-fold enhanced levels relative to control values. bFGF (1, 20, and 10 ng/ mL) enhanced neuronal viability (p < .05), and 10 ng/mL bFGF induced a maximal increase in live cells to 80% of controls. Axonal arborizations increased to 50% and dendritic arborizations to 75% of controls after 10 ng/mL bFGF (p< .05). bFGF in a dose of 20 ng/ mL enhanced axonal branching to 40% and dendrites in number and branching to 50% of controls (p< .05). bFGF (500 pg/mL, and 1 and 10 ng/mL) decreased enhanced AA (p < .05), and 10 ng/mL bFGF maximally reduced increased AA to two-fold enhanced values relative to controls. CONCLUSIONS: In vulnerable neurons, exposure to moderate lactic acidosis induces a process of cell injury with long latency. bFGF applied postinjury reduces the delayed neurodegeneration and may have neuroprotective efficacy in new therapeutic strategies to ischemia-induced cerebral injury. PMID- 9875917 TI - Effects of partial liquid ventilation with perfluorocarbons on pressure-flow relationships, vascular compliance, and filtration coefficients of isolated blood perfused rabbit lungs. AB - OBJECTIVES: The density of perfluorocarbons is almost twice that of blood. Therefore, we hypothesized that partial liquid ventilation with these fluids markedly affects pulmonary hemodynamics and filtration coefficients. To test these hypotheses we studied pressure-flow relationships, vascular compliances, capillary pressures, and filtration coefficients in normal and perfluorocarbon ventilated rabbit lungs. DESIGN: Controlled animal study with an ex-vivo isolated lung preparation. SETTING: Research laboratory for experimental anesthesiology at the Heinrich-Heine-University of Dusseldorf. SUBJECTS: Fourteen New Zealand White rabbits. INTERVENTIONS: The lungs were perfused under zone 3 flow conditions with autologous blood at various flow rates (50 to 250 mL/min, closed circuit, roller pump, 37 degrees C) and ventilated with 5% CO2 in air (positive end-expiratory pressure: 2 cm H2O, tidal volume: 10 mL/kg, respiratory rate: 30 breaths/min) without (control group, n=7) and with (n=7) perfluorocarbon administered intratracheally (15 mL/kg). MEASUREMENTS AND MAIN RESULTS: Pulmonary arterial, left atrial, and airway pressures, as well as blood reservoir volume (reflecting changes in pulmonary blood volume) and lung weight, were measured continuously. Inconsistent with our hypothesis, we found no significant differences between both groups in the slopes and intercepts of the pressure-flow relationships. There were no significant differences in capillary pressures determined by double occlusion (6.7+/-1.2 vs. 6.3+/-1.3 cm H2O for control group, p=.53), vascular compliances (0.51+/-0.10 vs. 0.47+/-0.09 mL/cm H2O for control group, p=.38), and filtration coefficients (0.33+/-0.06 vs. 0.37+/-0.07 mL/min/mm Hg/100 g wet weight for control group, p=.80, Mann-Whitney). CONCLUSIONS: Partial liquid ventilation with perfluorocarbons has no relevant effects on pulmonary filtration coefficients and global hemodynamic variables of isolated zone 3 lungs. These findings suggest that right ventricular afterload is not changed with partial liquid ventilation. It is likely, however, that intrapulmonary blood flow is redistributed toward less-dependent regions, although relevant global hemodynamic changes are absent during partial liquid ventilation. PMID- 9875918 TI - Nitric oxide production after acute, unilateral hydrochloric acid-induced lung injury in a canine model. AB - OBJECTIVE: To determine if acute unilateral lung injury induces only local or systemic inflammatory effects by measuring the production of nitric oxide (NO) and its metabolites (nitrites and nitrates) in the injured and the contralateral lung and the blood initially and 4 hrs after injury. DESIGN: Unilateral hydrochloric acid instillation in split lung intubated subject studied over time. SETTING: Animal physiology laboratory. SUBJECTS: Five mongrel dogs. INTERVENTIONS: Instillation of 10 mL of 0.1 N hydrochloric acid into one lung via a plastic catheter. Bronchoalveolar lavage (BAL) was done at 4 hrs. MEASUREMENTS AND MAIN RESULTS: Unilateral acid instillation did not alter systemic blood pressure or cardiac output, nor did it induce arterial hypoxemia. The BAL nitrite and nitrate level on the side of injury was higher than the control side (3.6+/ 1.36 vs. 1.5+/-1.58 mM, p < .05), and serum nitrites and nitrates levels also decreased from the levels before acid instillation levels (p < .05). Exhaled NO levels were measured only in three animals. The levels increased acutely on hydrochloric acid instillation from only the injured lung and returned to baseline over several minutes. However, the level of exhaled NO from the injured lung failed to increase 4 hrs after injury, despite the increase in BAL nitrites and nitrates. CONCLUSIONS: Acute unilateral lung injury in the dog results in increased NO production that is compartmentalized to the injured lung. The increase in exhaled NO after injury is transient and does not allow one to monitor the progress of lung injury. PMID- 9875919 TI - Ionized magnesium concentrations in critically ill children. AB - OBJECTIVE: To test the hypothesis that many critically ill children exhibit ionized hypomagnesemia despite having normal total magnesium (TMg) concentrations. DESIGN: A prospective, observational study with convenience sampling. SETTING: Pediatric and cardiovascular intensive care units of a large children's hospital. PATIENTS: Patients aged 1 day to 21 yrs admitted from January 1 to October 31, 1996. Patients with chronic renal failure or weight <3 kg were excluded. A group of healthy children involved in a school-based nutritional assessment study were also studied. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Sixty-seven patients (5.4+/-5.7 [SD] yrs) and 24 healthy children (10.84+/-0.93 yrs, p< .001) were studied. Plasma was assayed for ionized magnesium (IMg) using a blood analyzer. Forty (59%)/67 critically ill subjects had IMg concentrations <0.40 mmol/L, the lowest published normal value and the lowest value observed in our group of healthy children. Of these, 24 (60%)/40 had normal TMg concentrations. IMg was significantly (p=.00) lower in critically ill subjects than in the group of healthy children (0.37+/-0.10 mmol/L vs. 0.46+/ 0.03 mmol/L). IMg did not correlate strongly with ionized calcium (r2=0.49), albumin (r2=0.09), or pH (r2=0.18). CONCLUSION: Many critically ill children exhibit ionized hypomagnesemia with normal TMg concentrations. These children would not be recognized as magnesium-deficient based on routine TMg testing. Critically ill children exhibited significantly lower concentrations of IMg than a group of healthy children. PMID- 9875920 TI - A randomized, controlled trial of computerized physiologic trend monitoring in an intensive care unit. AB - OBJECTIVE: To assess whether the provision of computerized physiologic trend data could improve outcome in newborn infants requiring intensive care. DESIGN: Randomized, controlled trial, with subsidiary questionnaire studies. SETTING: Tertiary neonatal intensive care unit with 12 intensive care cots. PATIENTS: All infants admitted between January 1991 and September 1993 who were < or =32 wks gestation or >32 wks gestation, and ventilated for >4 hrs or asphyxiated. INTERVENTIONS: Randomization to one of four groups for first 7 days of life: A) no display of trend data; B) continuous display of trend data; C1) alternating 24 hr display of trend data, starting with display in first 24 hrs; and C2) alternating 24-hr display of trend data, starting with no display in first 24 hrs. MEASUREMENTS AND MAIN RESULTS: The short-term effects of monitoring on patient outcome was judged by volume of colloid given, number of blood gases taken, and by measurement taken from cranial Doppler ultrasound. Medium-term measures included time ventilated, time given supplemental oxygen, death, time to death or discharge, and cranial ultrasound at discharge. Long-term outcome was assessed by neurodevelopmental status at age 1 to 4 yrs of age. Staff and parent questionnaires assessed their respective attitudes to the introduction of this technology. None of the patient outcome measures, short-, medium-, or long-term, demonstrated any significant benefit from the provision of computerized physiologic trend monitoring. Staff questionnaires demonstrated an acceptance of the system and an improved understanding of neonatal physiology as a result of computerized physiologic trends. Parent questionnaires demonstrated increased anxiety caused by the system in 11% of parents, although only 1% of parents continued to have concerns if the system were able to help their child. CONCLUSIONS: A randomized, controlled trial was unable to demonstrate any benefit to patients resulting from the introduction of a computerized physiologic trend monitoring system. Benefits of the system have been recognized, however, in subsidiary studies, staff education, and research studies. PMID- 9875921 TI - Noninvasive management of pediatric neuromuscular ventilatory failure. AB - OBJECTIVE: To evaluate the use of mouth piece/nasal intermittent positive pressure ventilation (IPPV) as an alternative to intubation or to permit extubation for patients with primarily neuromuscular ventilatory impairment and no ventilator-free breathing ability. DESIGN: A case control study. INTERVENTIONS: Using a protocol in which oxyhemoglobin desaturation was prevented or reversed by the continuous use of noninvasive IPPV and manually and mechanically assisted coughing as needed, patients with neuromuscular ventilatory failure and no ventilator-free breathing ability were managed noninvasively or extubated to continuous use of noninvasive IPPV for ventilatory support on room air. MEASUREMENTS AND MAIN RESULTS: Four of ten patients who presented in acute ventilatory failure were managed without intubation, despite becoming dependent on continuous ventilator use. The six intubated patients were extubated successfully to continuous noninvasive IPPV once normal arterial oxygen saturation levels could be maintained on room air, despite their having no ventilator-free breathing ability. CONCLUSIONS: The use of inspiratory and expiratory aids can decrease the need for intubation for patients with neuromuscular ventilatory failure in the absence of significant lung disease. It can also permit extubation, despite the need for continuous ventilatory support and, thereby, decrease the need to resort to tracheostomy. PMID- 9875922 TI - Noninvasive cardiac output monitoring by aortic blood flow determination: evaluation of the Sometec Dynemo-3000 system. AB - OBJECTIVE: The Sometec Dynemo-3000 system allows the permanent measurement of descending aorta diameter by an echographic (A-scan) device and the blood flow velocity by a pulse Doppler velocimeter. The Dynemo-3000 then furnishes a new hemodynamic parameter, i.e., descending aortic blood flow (ABF), which is a fraction of the cardiac output (CO). We evaluate the ability of this system to measure the aortic diameter and to accurately detect ABF changes. DESIGN: A case study prospective trial. SETTING: A 24-bed medical intensive care unit of a 1,100 bed university hospital. PATIENTS: Twenty critically ill patients fully sedated, mechanically ventilated, and monitored by a pulmonary artery catheter. INTERVENTIONS: CO values determined by conventional thermodilution method (TD-CO) and ABF were recorded during the study, which included two initial baseline periods, a dobutamine infusion (5 microg/kg/min) interval of 30 mins, and a third baseline period. To assess the accuracy of A-scan, aortic diameter was measured by transesophageal echocardiography. The difference between echocardiography and A-scan was used to determine bias and precision for aortic diameter measurements. TD-CO and ABF variations were analyzed using Kruskal-Wallis and Wilcoxon tests. Association between TD-CO and ABF values was determined by calculating the linear correlation coefficient. The ability of ABF to detect a TD-CO >6.0 L/min and its variations >13% was analyzed by determination of sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values. MEASUREMENTS AND MAIN RESULTS: Aortic diameter measurements by A-scan and bidimensional methods were 23.0+/-2.8 mm (SD) and 24.2+/-2.7 mm, respectively. Bias and precision were 1.1 mm and 1.4 mm (95% confidence interval: -1.9 to 3.7), respectively. During the course of dobutamine infusion, we observed a significant increase of TD-CO mean value from 6.65+/-1.53 L/min to 9.30+/-2.5 L/min (p=.0008), and a parallel and significant increase in ABF mean value from 4.34+/-1.18 L/min to 5.70+/-1.63 L/min (p= .0029). Absolute TD-CO and ABF values had a correlation coefficient of 0.80. For detection of an increased TD-CO, PPV and NPV were 87% and 86%, respectively. For detection of TD-CO changes >13%, PPV and NPV were 80% and 94%, respectively. CONCLUSIONS: The Dynemo-3000 system is able to display the real aortic diameter, which is one of the most important components of this noninvasive ultrasonic technique. When compared with TD-CO, the ABF determination provided by this ultrasonic device constitutes a reliable noninvasive tool for estimating CO and tracking its changes. PMID- 9875923 TI - Has high-frequency ventilation been inappropriately discarded in adult acute respiratory distress syndrome? AB - OBJECTIVES: To review the basic physiologic principles that support the role for high-frequency ventilation (HFV) in acutely lung-injured patients, to critically assess clinical trial data in this area, and discuss why a metasummary is not feasible and a large-scale clinical trial is needed. DATA SOURCES: We searched a computerized database (MEDLINE) from 1976 to January 1997 using the text words "high-frequency ventilation" and "acute respiratory distress syndrome" to retrieve all relevant candidate articles. STUDY SELECTION: We retrieved all English language clinical studies conducted in tertiary care centers that employed HFV in adult acute respiratory distress syndrome (ARDS) patients. DATA EXTRACTION: Only prospective, randomized trials, cohort/case-control studies, and case series evaluating HFV vs. conventional mechanical ventilation in adult ARDS patients were included. DATA SYNTHESIS: We independently screened 3,166 articles on ARDS and 494 papers on HFV in our computer search. We checked reference lists and contacted experts in the field of mechanical ventilation in ARDS to ensure that no relevant studies had been missed. Only four articles met our inclusion criteria and were evaluated in detail. CONCLUSIONS: Current clinical studies are statistically under-powered and a metasummary is not feasible because of study quality, as well as lack of similar clinical end points and measures of magnitude of benefit. A large, multicenter trial should be initiated to define the role of HFV in the treatment of adult ARDS. PMID- 9875924 TI - Has the mortality of septic shock changed with time. AB - OBJECTIVES: To determine whether a systematic review of the literature could identify changes in the mortality of septic shock over time. DATA SOURCES: A review of all relevant papers from 1958 to August 1997, identified through a MEDLINE search and from the bibliographies of articles identified. DATA SYNTHESIS: The search identified 131 studies (99 prospective and 32 retrospective) involving a total of 10,694 patients. The patients' mean age was 57 yrs with no change over time. The overall mortality rate in the 131 studies was 49.7%. There was an overall significant trend of decreased mortality over the period studied (r=.49, p < .05). The mortality rate in those patients with bacteremia as an entry criterion was greater than that rate in patients whose entry criterion was sepsis without definite bacteremia (52.1% vs. 49.1%; chi2=6.1 and p< .05). The site of infection altered noticeably over the years. Chest related infections increased over time, with Gram-negative infections becoming proportionately less common. If all other organisms and mixed infections are included with the Gram-positives, the result is more dramatic, with these organisms being causative in just 10% of infections between 1958 and 1979 but in 31% of infections between 1980 and 1997. CONCLUSIONS: The present review showed a slight reduction in mortality from septic shock over the years, although this result should be approached with caution. The heterogeneity of the articles and absence of a severity score for most of the studies limited our analysis. Furthermore, there was an increasing prevalence of Gram-positive causative organisms, and a change of the predominant origin of sepsis from the abdomen to the chest. PMID- 9875925 TI - Lactic acidemia and bradyarrhythmia in a child sedated with propofol. AB - OBJECTIVES: To describe a severe adverse reaction in a child who received an infusion of propofol for sedation in the intensive care unit (ICU). To describe the management and further investigation of this patient and review similar published reports. DESIGN: Case report and literature review. SETTING: Community hospital ICU and tertiary pediatric ICU. PATIENT: Infant with upper respiratory obstruction secondary to an esophageal foreign body who required tracheal intubation and mechanical ventilation. INTERVENTIONS: Conventional cardiovascular and respiratory support. Continuous veno-venous hemofiltration (CVVH) and plasmapheresis. MEASUREMENTS AND MAIN RESULTS: The patient received a propofol infusion at a mean rate of 10 mg/kg/hr for 50.5 hrs. He developed lipemia and green urine and subsequently, a progressive severe lactic acidemia and bradyarrhythmias unresponsive to conventional treatment. These abnormalities resolved with CVVH. He was encephalopathic and developed liver and muscle necrosis histologically compatible with a toxic insult. Examination of homogenized muscle tissue demonstrated a reduction in cytochrome C oxidase activity. There was no evidence of systemic infection or underlying metabolic disease. He eventually recovered completely. CONCLUSION: Propofol has been associated with severe adverse reactions in children receiving intensive care. The biochemical and histologic abnormalities described in this patient may guide further investigation. We advise against prolonged use of propofol for sedation in children. PMID- 9875926 TI - Financial disclosure is needed at the lectern, as well as in print. PMID- 9875927 TI - Lipid emulsions with different phospholipid/triglyceride ratio in critically ill septic and traumatic patients. PMID- 9875928 TI - Noninvasive ventilation in children with respiratory failure. PMID- 9875929 TI - The prognostic importance of the ossific nucleus in the treatment of congenital dysplasia of the hip. AB - Ischemic necrosis of the femoral head occurring after the treatment of congenital dysplasia of the hip can negatively affect the long-term prognosis of the involved hip. Some investigators have suggested that the presence of the ossific nucleus of the femoral head at the time of closed or open reduction is associated with a lower rate of ischemic necrosis. This finding, if verified, could lead to a delay in the treatment of a dislocated hip until ossification of the femoral head has begun, which may be well after the age when the patient has started to walk. We conducted a computerized search of the medical records at our two tertiary-care children's hospitals to identify all patients with congenital dysplasia of the hip who had had a closed or open reduction between January 1, 1979, and December 31, 1993. One hundred and twenty-four patients (153 hips) who satisfied the criteria for inclusion were identified. The ossific nucleus was present in ninety hips and absent in sixty-three. Closed reduction was used in 112 hips and open reduction, in forty-one. Ischemic necrosis was identified in five hips (3 percent): four (6 percent) of the sixty-three hips that did not have an ossific nucleus and one (1 percent) of the ninety hips that had an ossific nucleus at the time of the reduction. With the numbers available for study, we could not detect a difference between these two groups. The age at reduction (p > 0.99), the method of reduction (p = 0.611), previous treatment with a Pavlik harness (p = 0.592), the use of preliminary traction (p = 0.602), concomitant procedures (p > 0.99), and a failure of the primary closed reduction (p = 0.579) were not associated with the development of ischemic necrosis after reduction. In our analysis of patients who were managed over a fifteen-year period, the data did not support the hypothesis that the presence of an ossific nucleus at the time of reduction of a congenitally dislocated hip is associated with a lower prevalence of ischemic necrosis of the femoral head. Sound operative principles dictate that operative reduction of a congenitally displaced hip should be performed when the child can be safely placed under anesthesia and without regard to the presence or absence of the ossific nucleus. PMID- 9875930 TI - Dural tears secondary to operations on the lumbar spine. Management and results after a two-year-minimum follow-up of eighty-eight patients. AB - We reviewed the results of acute management of patients who had sustained a dural tear during an operation on the lumbar spine, and we attempted to determine the long-term sequelae of this complication. In the five years from July 1989 to July 1994, 641 consecutive patients had a decompression of the lumbar spine, performed by the senior one of us; of these patients, eighty-eight (14 percent) sustained a dural tear, which was repaired during the operation. The duration of follow-up ranged from two to eight years (average, 4.3 years). Postoperative management consisted of closed suction wound drainage for an average of 2.1 days and bed rest for an average of 2.9 days. Of the eighty-eight procedures that resulted in a dural tear, forty-five were revisions; these revisions were performed after an average of 2.2 previous operations on the lumbar spine, all of which resulted in a scar adherent to the dura. Only eight patients had headaches related to the spinal procedure and photophobia in the postoperative period; these symptoms resolved in all but two patients, both of whom had had a revision operation. Each of the two patients had symptoms of a persistent leak of spinal fluid and needed a reoperation for repair. Overall, seventy-six patients had a good or excellent result and twelve had a poor or satisfactory result with some residual back pain. One patient had arachnoiditis, and another had symptoms of viral meningitis one month postoperatively. A dural tear that occurs during an operation on the lumbar spine can be treated successfully with primary repair followed by bed rest. Such a tear does not appear to have any long-term deleterious effects or to increase the risk of postoperative infection, neural damage, or arachnoiditis. Closed suction wound drainage does not seem to aggravate the leak and can be used safely in the presence of a dural repair. PMID- 9875931 TI - Monteggia fractures in adults. AB - The records concerning ten consecutive years of experience with Monteggia fractures in adult patients at a level-one trauma center were retrospectively reviewed. Forty-eight patients who had been followed for a minimum of two years (average, 6.5 years; range, two to fourteen years) were identified. There were twenty-five women and twenty-three men, and the average age was fifty-two years (range, eighteen to eighty-eight years). According to the classification of Bado, there were seven type-I, thirty-eight type-II, one type-III, and two type-IV injuries. Twenty-six patients (68 percent) who had a Bado type-II fracture had an associated fracture of the radial head; ten of these patients also had a fracture of the coronoid process as a single large fragment. The ulna was fixed with a tension band-wire construct supplemented with screws in three patients (all of whom had a Bado type-II fracture). An ulnar diaphyseal fracture was fixed with an intramedullary Steinmann pin in one patient. The remaining patients had fixation with a plate and screws. The fracture of the radial head was treated with either complete or partial excision of the fragments in twelve patients (with replacement with a silicone prosthesis in two), open reduction and internal fixation in ten patients, and no intervention in four patients. Nine patients, all of whom had a Bado type-II fracture, needed a reoperation within three months after the initial operation; five had revision of a loose ulnar fixation device, three had resection of the radial head, and one had removal of a wire that had migrated from the radial head into the elbow articulation. Other important complications included proximal radioulnar synostosis in three patients, ulnar malunion in three, posterolateral rotatory instability of the ulnohumeral joint in one, and instability of the distal radioulnar joint in one. At the most recent follow-up examination, which was performed after all of the reoperations and reconstructive procedures had been done, the average score according to the system of Broberg and Morrey was 86 points (range, 15 to 100 points). The result was excellent for eighteen patients, good for twenty-two, fair for two, and poor for six. Six of the eight patients who had an unsatisfactory (fair or poor) result had had a Bado type-II fracture with a concomitant fracture of the radial head. These unsatisfactory results were related to a malunited fracture of the coronoid process in two patients, a proximal radioulnar synostosis in one, a malunited fracture of the coronoid process and a proximal radioulnar synostosis in one, a malunion of the ulna in one, and painfully restricted rotation of the forearm after operative fixation of a comminuted fracture of the radial head in one. The other two unsatisfactory results were in a patient who had had a Bado type-I fracture and in one who had had a Bado type-IV fracture. The results of the present series are much better than those reported in most earlier studies, suggesting that stable anatomical fixation of the ulnar fracture (including associated fracture fragments of the coronoid process) with a plate and screws inserted with use of current techniques of fixation leads to a satisfactory result in most adults who have a Monteggia fracture. The posterior (Bado type-II) fracture is the most common type of Monteggia fracture in adults. Problems with the elbow related to fractures of the coronoid process and the radial head, which are common with Bado type-II Monteggia fractures, remain the most challenging elements in the treatment of these injuries. PMID- 9875932 TI - The chondrogenic potential of human bone-marrow-derived mesenchymal progenitor cells. AB - Mesenchymal progenitor cells provide a source of cells for the repair of musculoskeletal tissue. However, in vitro models are needed to study the mechanisms of differentiation of progenitor cells. This study demonstrated the successful induction of in vitro chondrogenesis with human bone-marrow-derived osteochondral progenitor cells in a reliable and reproducible culture system. Human bone marrow was removed and fractionated, and adherent cell cultures were established. The cells were then passaged into an aggregate culture system in a serum-free medium. Initially, the cell aggregates contained type-I collagen and neither type-II nor type-X collagen was detected. Type-II collagen was typically detected in the matrix by the fifth day, with the immunoreactivity localized in the region of metachromatic staining. By the fourteenth day, type-II and type-X collagen were detected throughout the cell aggregates, except for an outer region of flattened, perichondrial-like cells in a matrix rich in type-I collagen. Aggrecan and link protein were detected in extracts of the cell aggregates, providing evidence that large aggregating proteoglycans of the type found in cartilaginous tissues had been synthesized by the newly differentiating chondrocytic cells; the small proteoglycans, biglycan and decorin, were also detected in extracts. Immunohistochemical staining with antibodies specific for chondroitin 4-sulfate and keratan sulfate demonstrated a uniform distribution of proteoglycans throughout the extracellular matrix of the cell aggregates. When the bone-marrow-derived cell preparations were passaged in monolayer culture as many as twenty times, with cells allowed to grow to confluence at each passage, the chondrogenic potential of the cells was maintained after each passage. PMID- 9875933 TI - Results of the modified Sauve-Kapandji procedure in the treatment of chronic posttraumatic derangement of the distal radioulnar joint. AB - We reviewed the results of a modified Sauve-Kapandji procedure with tenodesis of the flexor carpi ulnaris to the carpus in eighteen patients who had chronic derangement of the distal radioulnar joint. There were fourteen men and four women. The mean supination of the forearm had improved from 16 degrees (range, 0 to 75 degrees) preoperatively to 76 degrees (range, 40 to 90 degrees) at the time of the latest follow-up, and the mean pronation had improved from 42 degrees (range, 0 to 80 degrees) preoperatively to 81 degrees (range, 60 to 90 degrees) at the time of follow-up. Pain relief was satisfactory, and the mean grip strength had improved from 36 percent of that on the unaffected side preoperatively to 73 percent at the time of follow-up. One patient had moderate pain over the ulnar stump associated with residual volar instability of the proximal ulnar segment, and he had a tenodesis of the extensor carpi ulnaris as a second procedure. Another patient had mild instability of the stump only after he had a second operation, which was an excision of a bone mass (ossification) in the resected area. The ulnar stump was stable in sixteen patients. Eight of the eleven patients who had performed heavy manual labor before the injury were able to return to work full-time without restrictions. According to a modification of the wrist-scoring system of the Mayo Clinic, at a mean of four years and two months (range, two years to eight years and four months), six patients had an excellent result; seven, a good result; four, a fair result; and one, a poor result. On the basis of our findings, we believe that the index operation is an excellent salvage procedure for the treatment of chronic posttraumatic derangement of the distal radioulnar joint, especially when nonoperative treatment has been unsuccessful and rotation of the forearm is severely limited. PMID- 9875934 TI - Total knee arthroplasty in morbidly obese patients. AB - We reviewed the clinical outcomes of fifty primary total knee arthroplasties that had been performed with cement in forty patients who were considered morbidly obese (a Quetelet index of more than forty). These results were compared with those of 1768 similar procedures, performed during the same time-period by the same surgeon, in 1539 patients who were not morbidly obese (controls). At a mean of approximately five years postoperatively, there was a significant difference between the morbidly obese patients and the control group with regard to the knee and functional scores (84 and 53 points compared with 92 and 67 points; p < 0.00005 for both scores). No significant difference was detected, with the numbers available, with regard to the range of motion or the radiographic score (p = 0.77). The rate of perioperative complications was significantly higher in the morbidly obese patients (p < 0.00005). Of the fifty knees in these patients, eleven (22 percent) had a wound complication, five (10 percent) had an infection, and four (8 percent) had an avulsion of the medial collateral ligament. The five infections developed within twenty weeks after the operation, and three were associated with a wound complication. In comparison, thirty-five (2 percent) of the 1768 knees in the control group had a wound complication, eleven (0.6 percent) had an infection, and none had an avulsion of the medial collateral ligament. We concluded that total knee arthroplasty in morbidly obese patients can be successful but is associated with an increased rate of perioperative complications, including problems with wound-healing, infection, and avulsion of the medial collateral ligament. Alterations in the operative technique for soft tissue closure and protection of the medial collateral ligament have decreased the rates of complications related to wound-healing and the medial collateral ligament. PMID- 9875935 TI - The Harris Design-2 total hip replacement fixed with so-called second-generation cementing techniques. A ten to fifteen-year follow-up. AB - We analyzed the clinical results of 195 Harris Design-2 total hip replacements performed with so-called second-generation cementing techniques in 166 consecutive patients who had osteoarthrosis. The mean age of the patients at the time of the replacement was sixty-seven years and nine months (range, thirty-one to eighty-nine years). Forty-eight patients (fifty-four hips) died before the time of the latest follow-up, but the implants were apparently functioning well at the time of death. Three patients (four hips) were lost to follow-up. Five patients (five hips; 3 percent) had a revision because of aseptic loosening of the acetabular or femoral component, or both, that was related to wear-induced osteolysis. The mean Harris hip score for the 131 hips that were available at the latest follow-up examination at a mean of twelve years (range, ten to fifteen years) after the operation was 89 +/- 10 points. On the basis of the Harris hip score, seventy-six hips had an excellent result, thirty-four had a good result, fifteen had a fair result, and six had a poor result at the latest follow-up examination. Radiographically, twelve (9 percent) of the 131 acetabular components and three (2 percent) of the 131 femoral components were probably or definitely loose. At a mean of twelve years, 186 (97 percent) of 191 Harris Design-2 implants were in situ or had been in situ at the time of the patient's death. PMID- 9875936 TI - The role of the acetabular labrum and the transverse acetabular ligament in load transmission in the hip. AB - We performed a biomechanical study of seventeen hip joints in the pelves of nine cadavera in order to assess the role that the acetabular labrum and the transverse acetabular ligament play in load transmission. The distribution of contact area and pressure between the acetabulum and the femoral head was measured with the hip in four different conditions: intact (seventeen hips), after removal of the transverse acetabular ligament (eight hips), after removal of the entire labrum (nine hips), and after removal of both the transverse acetabular ligament and the labrum (seventeen hips). The hip joint was loaded in simulated single-limb stance, and the measurements were made with use of pressure sensitive film. A peripheral distribution of load was seen in the intact acetabula. This pattern was altered only minimally after removal of the transverse acetabular ligament or the labrum, or both. When both of these structures were removed, the only significant change was a decrease in the maximum pressure in the posterior aspect of the acetabulum (p = 0.02). No significant changes were detected with regard to the contact area, load, mean pressure, or maximum pressure in the anterior or superior aspect of the acetabulum under any testing condition. PMID- 9875937 TI - Delayed onset of anterior tibial compartment syndrome in a patient receiving low molecular-weight heparin. A case report. PMID- 9875938 TI - Late foreign-body reaction to an intraosseous bioabsorbable polylactic acid screw. A case report. PMID- 9875939 TI - The healing and regeneration of articular cartilage. PMID- 9875940 TI - Evolution of the present status of orthopaedic surgery fellowships. PMID- 9875941 TI - Impact of fellowships on orthopaedic surgery residencies, patient care, and orthopaedic practice. Part I. PMID- 9875942 TI - Impact of fellowships on orthopaedic surgery residencies, patient care, and orthopaedic practice. Part II. PMID- 9875943 TI - Issues with accreditation and certification of orthopaedic surgery fellowships. PMID- 9875944 TI - Orthopaedic surgery fellowships: a ten-year assessment. Subspecialty training during residency and the ideal academic fellowship. PMID- 9875945 TI - Orthopaedic surgery fellowships: a ten-year assessment. Sponsoring and funding fellowships: a private-practice model. PMID- 9875946 TI - Orthopaedic surgery fellowships: a ten-year assessment. Fellowship accreditation and certification methodologies of other specialties. PMID- 9875947 TI - Current concepts review. Subacromial impingement syndrome (79-A: 1854-1868, Dec. 1997) PMID- 9875948 TI - Current concepts review. Subacromial impingement syndrome (79-A: 1854-1868, Dec. 1997) PMID- 9875949 TI - Current concepts review. Subacromial impingement syndrome (79-A: 1854-1868, Dec. 1997) PMID- 9875950 TI - Elevated peak plantar pressures in patients who have Charcot arthropathy (80-A: 365-369, March 1998) PMID- 9875951 TI - Evolutionary psychopathology: why isn't the mind designed better than it is? AB - This paper outlines some of the basic issues in evolutionary thinking when applied to psychopathology. It aims to give some background and context to the papers that follow. Particular attention is given to the way evolution is constrained by previous design features, the importance of trade offs between advantages and disadvantages of different traits, the way social dilemmas and conflicts have shaped the evolution of human motivational systems, and the psychology that supports them. The paper concludes with a plea to avoid thinking of evolutionary approaches as a reductionist science, when the focus needs to be on the way evolution theory informs the study of the social-biological interface. PMID- 9875952 TI - Evolutionary epidemiology and manic depression. AB - The reformulation of epidemiological prevalence rates as evolutionary frequency rates puts medical genetics within an explicit framework of Darwinian theory. Yet an enduring and still current assumption of genomic medicine is that genes associated with disease are necessarily maladapted. Indeed, it seems it could hardly be otherwise. However, evolutionary epidemiology has begun to uncover important and surprising counter-exemplary case-studies. Thus, the present aim is to first outline this emerging sub-discipline of 'evolutionary epidemiology'. Then, a major psychopathological syndrome--manic-depression--is examined in some detail within the purview of evolutionary epidemiology. Its medical genetics are those of an adaptive polymorphism in the human genome. Hence, genes associated with what is now a major public health problem accrued as they conferred selective advantage in phylogeny. Why should manic-depressive etiogenes have been selected? A preliminary anatomic-functional model, assembled from facts of human paleoneuropsychiatry, more adequately contextualises manic-depressive genomics and phenotypy. In this model, manic-depression finds its heuristic origins in a hierarchy of behavioural strategies stabilised in phylogeny and embedded at serial levels in the brain (Hawk-Dove ESS). A proportion of the population has variant genotypy which appears to have been favoured in social competition phylogenetically but express more pathogenic phenotypy in the current environment. The paper closes with a brief consideration of clinical practices and ethical issues as alternative considerations emerge with the syndrome recast in a more positive Darwinian light. PMID- 9875953 TI - Emotional disorders in evolutionary perspective. AB - Understanding emotional disorders requires understanding the evolutionary origins and functions of normal emotions. They are special states, shaped by natural selection to adjust various aspects of the organism in ways that have tended to give a selective advantage in the face of the adaptive challenges characteristic of a particular kind of situation. They are designed to maximize reproductive success, not happiness. Negative emotions such as anxiety and low mood are not disorders, but, like the capacity for pain, evolved defences. Excessive anxiety or low mood is abnormal, but we will not have confidence about what is excessive until we understand their functions better than we do. Emotional disorders arise often from social emotions because of the conflicts inherent in social life, and because of the strategic advantages of demonstrating commitments to follow through on threats and promises. An evolutionary understanding of individuals in terms of their relationship strategies and the social emotions offers great promise for psychotherapists. PMID- 9875954 TI - Ethological strategies for defence in animals and humans: their role in some psychiatric disorders. AB - Ethological strategies for defence in animals and humans are expressed as either aggression or flight behaviour. Aggression is employed by animals during intraspecific competition for resources, mate, territory and acquiring and maintaining social status. It also disperses individuals throughout the biotope. Flight behaviour is used to avoid a source of danger or harm, has both dynamic arid static forms, is phylogenetically very old and takes precedence over all other activities including social behaviour. Animals exposed to inescapable threats or attacks exhibit a characteristic defensive strategy, arrested flight, which consists of gaze-avoidance or cut-offs, cryptic postures such as immobility and covert surveillance of their surroundings. Arrested flight also occurs in social encounters when submission fails to reduce attacks, and in prey animals when escape from a predator is hampered. Ethological studies show that during interviews, depressed patients exhibit a pattern of non-verbal behaviour having all the hallmarks of arrested flight. Cut-off behaviour, which seeks to reduce the input of flight-evoking stimuli is especially evident in these patients but takes an extreme form, i.e. eye closure, in the gaze-profiles of paranoid patients. It is proposed that cut-offs always denote the presence of incipient flight and that arrested flight is a 'last measure' defensive strategy in response to inescapable proximal threat. It can arise in humans whenever their escape routes are hampered and characterizes the behaviour of patients suffering from depression. As in animals, different pathways may lead to arrested flight in humans. In humans, defensive mechanisms also operate at the mental level through putative ego defences, the psychological function of which is to preserve self esteem by hindering the access of disturbing emotional material into awareness. It is suggested that they function ethologically as mental cut-offs analogous to the behavioural cut-offs in animals. PMID- 9875955 TI - The evolved basis and adaptive functions of cognitive distortions. AB - This paper explores common cognitive distortions from the perspective of evolutionary psychology. It is suggested that cognitive distortions are natural consequences of using fast track defensive algorithms that are sensitive to threat. In various contexts, especially those of threat, humans evolved to think adaptively rather than logically. Hence cognitive distortions are not strictly errors in brain functioning and it can be useful to inform patients that 'negative thinking' may be dysfunctional but is a reflection of basic brain design and not personal irrationality. The evolved nature of cognitive distortions has been implicit in cognitive therapy from its early days (Beck, 1963; Ellis, 1962) but has not been fully articulated in what is now known about evolved mental processes. Many forms of cognitive distortion can be seen to use the (previously) adaptive heuristic of better safe than sorry. PMID- 9875956 TI - The adaptive function of mood change. AB - It is useful to imagine an 'agonistic strategy set', containing the two alternative and mutually incompatible strategies of escalation and de-escalation. The strategy set is accessed by loss, threat or some other form of 'ranking stress'. It is further suggested that ranking stress is dealt with relatively independently at three levels of the brain/mind, so that an agonistic strategy set is deployed at each of the three levels. Normally, all three levels escalate or de-escalate in unison, but sometimes lower level de-escalation is associated with middle or higher level escalation, and then the resolution of agonistic situations is delayed and psychopathology may be recognized. PMID- 9875957 TI - Prevalence differences in depression among males and females: are there evolutionary explanations? AB - Differences in male-female prevalence rates of depression are interpreted in evolutionary perspective. Three evolutionary hypotheses are evaluated: (a) depression represents an evolved strategy to deal with adverse social interactions, particularly among partners, (b) depression has self-preservative and manipulative features, and (c) depression is a consequence of failing to achieve biological goals. While these hypotheses are not necessarily mutually exclusive, evolutionary explanations often suffer from not specifying which hypothesis is primary and which are secondary. We argue that failing to achieve biological goals is primary and that evolved strategies and self-preservative and manipulative behaviour are secondary. Findings pointing to depression contributing features of male-female differences, asymmetries in male-female relationships and male-female differences associated with reproduction are discussed to support our view that evolutionary explanations can adequately account for a significant percentage of the reported prevalence differences. PMID- 9875958 TI - The brain and communication are basic for clinical human sciences. AB - This article focuses on basic concepts modelled on medical science for the human sciences. This reformulates problems experienced by people who consult counsellors, psychologists, psychiatrists, social workers and other human science clinicians as located in the brain. These troubles can be described as difficulties in social life and human communication, on the one hand, and as varied brain physiology, on the other. These problems and their solutions are not traditionally biological but restrictive views of biology need modification as the brain is obviously the central organ for not only the medical specialities of neurology and psychiatry but for all professionals concerned with social interactions. The human genome determines the brain of each person: each such brain constitutes the latest iteration of ancestral genomes that include species precursor to humans and primates, extending back to unicellular life forms. The genome that determines the human brain confers remarkable flexibility or learning potential. Yet many factors influence what is learned and experienced. Understanding this entails comparing and contrasting humans and non-humans. The genomes of chimpanzees and gorillas differ little from that of humans so most basic plans determining behaviour must be shared. Yet contrasted to these animals, the human brain is three times greater in mass and the human cerebral cortex has four times more area. This increased brain correlated with more social interaction; humans are the story-telling animal, producing, consuming and otherwise using small and large tales intensely and incessantly. But do communicational features resemble each other across species? Commnunicational propensity states in humans compare to those of non-humans. So do those of normal people and psychiatric patients. Psychiatry's efforts at systematic description and nosology provided guidelines to the ethology of ancient but still active communicational propensity states called PSALICs (Gardner, 1988). This double acronym refers to their normal function and prehuman origins; they are defined from a three-legged base in that each exists in psychiatric patients, normal people and non-human animals. This article describes the following psalics: alpha, audience, in-group omega, mating, nurturant, nurturance-eliciting, out group omega and spacing-avoidant. The article describes psalics' varied expression in people. This basic science formulation and an across-species comparisons approach has implications for treating patients and clients. PMID- 9875959 TI - Evolutionary kinship therapy: basic principles and treatment implications. AB - The idiom of kinship is a powerful one that stretches across many levels of social behaviour (van den Berghe, 1979), and Daly, Salmon & Wilson (1997) recently outlined the basic principles of a comprehensive, evolution-based kinship psychology. Their approach merges traditional kinship theory, Hamilton's inclusive fitness model and the broader realm of evolutionary psychology into an exciting and provocative call to arms. They address biological, psychological (viz. fictive kinship) and kin-like levels of analysis, and they argue for a relationship-specific kinship psychology that deals with the particular demands of being a mother, father, mate, offspring or grandparent. Our particular approach to kinship psychology has focused primarily on the distinction between biological kinship and psychological kinship (Bailey, 1988; Bailey & Wood, 1993; Nava & Bailey, 1991), and more recently on the role of kin-like relations in psychotherapy and other social contexts (Bailey, 1997a; Wood, 1997). The kin-like category is especially noteworthy in modern industrialized countries where acquaintanceships and stranger-to-stranger interactions often predominate over biological and psychological kinships (Ahern & Bailey, 1997). PMID- 9875960 TI - The sexual competition hypothesis for eating disorders. AB - A hypothesis is presented for eating disorders, based on Darwinian theory, that contends that these syndromes together with the phenomenon of the pursuit of thinness are manifestations of female intra-sexual competition. It is suggested that eating disorders originate in the human female's psychological adaptation of concern about physical attractiveness which is an important component of female 'mate attraction' and 'mate retention' strategies. It is argued that present-day environment of Western countries presents a range of conditions which have led to the overactivation or the disruption of the archaic female sexual strategy of maximizing 'mate value'. The present hypothesis deals with the ultimate level of causation and is therefore compatible with a range of theories of proximate causation. Although the present hypothesis is not directly testable, it makes predictions that are testable and refutable. Finally it is suggested that the sexual competition hypothesis has more explanatory power than existing evolutionary theories of eating disorders. PMID- 9875961 TI - Lymphatic mapping in the management of melanoma in children. AB - Lymphatic mapping allows the surgeon to identify and remove the first draining (sentinel) lymph node from a primary melanoma with minimal morbidity. The procedure facilitates accurate staging and identification of patients in need of additional therapy. We used lymphatic mapping and sentinel lymph node biopsy in two children with melanoma. Both patients had evidence of metastatic melanoma in their sentinel lymph nodes and underwent regional lymphadenectomy. Malignant melanoma and atypical pigmented lesions in children remain diagnostically challenging for the pathologist and clinician. Misdiagnoses occur, and the correct interpretation of a melanocytic tumor is too often made only after recurrence or metastasis has occurred. The use of lymphatic mapping facilitates accurate staging and identifies children in whom additional therapy may be indicated. In addition, it can assist in the assessment of the biologic potential of a difficult lesion. PMID- 9875962 TI - Cutaneous manifestations of Henoch-Schonlein purpura in young children. AB - We evaluated the anatomic distribution and pattern of skin involvement in 155 pediatric patients with Henoch-Schonlein purpura treated in our center over the last 20 years. Of these, 120 (77.4%) presented with cutaneous signs; only 10% of them had no leg involvement. Seven patients (4.5%) had edema of the hands, feet, or face. The mean age of this subgroup was 9.2+/-2.2 months, compared to 6.8+/ 2.3 years for the entire group (p < 0.05). Skin biopsies were performed in 18 of these patients, including the 7 with edema, to confirm the clinical diagnosis. All biopsy specimens showed leukocytoclastic vasculitis, with IgA deposits in two and C3 deposits in one. It is important that clinicians keep in mind that an atypical presentation of Henoch-Schonlein purpura is not unusual. PMID- 9875963 TI - Mucocutaneous manifestations in children with HIV infection and AIDS. AB - Mucocutaneous diseases are more frequent in HIV/AIDS-infected children than in the normal population. We analyze mucocutaneous disorders with atypical presentations in a large population of HIV-infected children, with or without full-blown AIDS, compared to a population of HIV seroreverted children. The majority of these cutaneous disorders have an infectious etiology and their frequency is related to the degree of deterioration of the immune system. Some diseases commonly observed in adults are rare in children; neoplasms are an exception. PMID- 9875964 TI - Chronic recurrent multifocal osteomyelitis preceding pyoderma gangrenosum and occult ulcerative colitis in a pediatric patient. AB - Chronic recurrent multifocal osteomyelitis (CRMO) is a clinicopathologic entity of unknown origin mainly affecting children and young adults. It shares many clinical features with pyoderma gangrenosum (PG), an uncommon ulcerating inflammatory disorder of the dermis and underlying subcutaneous fat. Both conditions are chronic and relapsing, and have been associated with inflammatory bowel disease (IBD). The diagnoses are made by exclusion. Histology is nonspecific and cultures are negative. There has been only one previous report of both conditions occurring in the same patient. Here we describe the second case, a 12-year-old girl who developed pyoderma gangrenosum in direct continuity to an underlying CRMO lesion of the right anterior tibia. Occult ulcerative colitis (UC) was subsequently discovered. PMID- 9875965 TI - Childhood discoid lupus erythematosus: a report of two cases. AB - Discoid lupus erythematosus (DLE) is an uncommon disease in childhood. We present two patients initially diagnosed as impetigo and photosensitive eczema with impetigo, respectively, who failed to respond to topical and systemic antistaphylococcal agents and in whom a diagnosis of discoid lupus erythematosus subsequently became apparent. PMID- 9875966 TI - Congenital lower lip pits (Van der Woude syndrome): presentation of 10 cases. AB - Congenital lower lip pits are a rare developmental malformation of the lower lip. Clinically they present as bilateral depressions in the vermilion zone of the lower lip. It is important to be aware of this disorder because lower lip pits have also been reported in a variety of other congenital disorders and are associated with other malformations. Ten cases of this entity are presented. Nine of the 10 patients are members of two related families. The clinical and pathologic picture as well as some of the genetic and therapeutic aspects of this peculiar condition are discussed. PMID- 9875967 TI - Childhood vulval lichen sclerosus in a patient with ectodermal dysplasia and uncombable hair. AB - An 11-year-old girl was diagnosed as having ectodermal dysplasia, with abnormalities of her nails, teeth, and hair noted from infancy. Her hair was unruly and investigations confirmed the changes seen in "cheveux incoiffables" the uncombable hair syndrome. She also had atopy requiring treatment to control asthma and eczema. When she was 8 years old she developed vulval lichen sclerosus, which was controlled easily with early treatment. The coexistence of these conditions is discussed. PMID- 9875968 TI - Papulonecrotic tuberculid in a 2-year-old girl: with emphasis on extent of disease and presence of leucocytoclastic vasculitis. AB - The clinicopathologic features of childhood papular necrotic tuberculid (PNT) have been detailed in 10 patients. PNT is characterized by symmetric, acral papular lesions that undergo necrosis. Despite widespread cutaneous involvement, the number of lesions is usually sparse. The extensor aspects of the limbs are usually involved and trunk involvement is uncommon. Vasculitis, an integral component of PNT, has been documented only once in childhood PNT. We describe PNT in a 2-year-old girl in whom a multitude of lesions were present all over the body, including unusual involvement of flexor surfaces of the limbs, trunk, perineum, and vulva. Biopsied tissue revealed the characteristic features of PNT, including leucocytoclastic vasculitis. There was prompt response to antituberculous therapy. Based on the findings in this patient, we believe that the clinical spectrum of this eminently treatable disease in children must be expanded in terms of distribution and number of lesions to include extensive limb and trunk, perineal, and vulval involvement; the histopathologic spectrum of childhood PNT must include leucocytoclastic vasculitis; and adult and childhood PNT share common histopathologic features, including a common cellular immunohistochemical profile, thereby suggesting a common pathogenesis. PMID- 9875969 TI - Measles virus was present in the inner cell of the acrosyringium in the skin rash. AB - A case of measles in a 26-year-old Japanese man is reported. A skin specimen taken on the third eruptive day from a maculopapular eruption on his chest was immunohistopathologically and electron microscopically examined using a rabbit polyclonal antibody against the nucleocapsid protein of the measles virus. The measles virus antigen was found in the inner cells of the acrosyringium and hair follicles. The measles virus nucleocapsid was electron microscopically identified in the nuclei of the inner cells of the acrosyringium. The findings suggest that the sweat from skin lesions might contain the measles virus. PMID- 9875970 TI - Van der Woude syndrome. A case report. AB - We describe several members of a family with Van der Woude syndrome, a genetic and congenital malformation syndrome with autosomal dominant inheritance and 70% to 80% penetrance with variable expressivity. It is characterized by clinical signs localized to the face, such as bilateral or unilateral pits on conical elevations in babies or extensive depressions in adults, both in the vermilion border of the lower lip, with cleft lip, with or without cleft palate and uvula. Small accessory or heterotopic salivary glands empty into sinuses or fistulas in the lips. This eight member family had various clinical signs of the condition. All had cleft lip and palate. We studied the major characteristics of the eight patients and describe histopathologic and immunohistochemical features. PMID- 9875971 TI - Onychomycosis in children: prevalence and management. AB - Onychomycosis in children is often accompanied by tinea pedis and a family history of onychomycosis. The prevalence of onychomycosis in children is substantially lower than that of adults; therefore it is important to confirm the clinical diagnosis. The most common presentation of onychomycosis is the distal and lateral subungual type. The organism most commonly isolated in North America is Trichophyton rubrum. Oral antifungal therapy is required, especially when the onychomycosis is of moderate to severe intensity, with nail matrix involvement. The new oral antifungal agents itraconazole, terbinafine, and fluconazole are being increasingly used for the treatment of onychomycosis. Review of the literature suggests that these agents are effective and safe in managing onychomycosis in children. The short duration of therapy required with these drugs should help improve compliance. The data suggest that the new oral antifungal agents have a role in the treatment of onychomycosis in children. Further experience will help us better position these drugs when evaluating the management of onychomycosis in children. PMID- 9875973 TI - What syndrome is this? Uncombable hair syndrome (Pili trianguli et canaliculi). PMID- 9875972 TI - Itraconazole oral solution for the treatment of onychomycosis. AB - The newer antifungal agents are increasingly being used for the treatment of onychomycosis. Tinea unguium is uncommon in children. In young children who are unable or unwilling to swallow capsules, the alternative to itraconazole capsules may be the oral solution. The pharmacokinetics of the itraconazole capsule when it is broken open and the contents sprinkled onto food has not been reported. Two sisters, ages 8 and 11 years, presented with fingernail dystrophy that was confirmed to be onychomycosis due to Trichophyton rubrum and T. soudanense, respectively. The patients were reluctant to swallow capsules and were treated with itraconazole pulse therapy using the oral solution, 3 mg/kg/day. The treatment regimen consisted of two pulses, each 1 week long, with a 3-week period between pulses. Both patients were clinically and mycologically cured with no adverse effects. Itraconazole oral solution (3 mg/kg/day) given as pulse therapy may be a consideration in the treatment of onychomycosis in some children. PMID- 9875974 TI - Thickened palms and soles in a toddler. PMID- 9875975 TI - The subcuticular "spider" stitch: a simple solution to suture breakage and patient discomfort in long incisions. PMID- 9875976 TI - Treatment of tufted angioma with interferon alfa: role of bFGF. PMID- 9875978 TI - Urticaria pigmentosa mistaken for child abuse. PMID- 9875977 TI - Extreme rises in serum alkaline phosphatase in children with atopic dermatitis after intervention treatment with cyclosporin A. PMID- 9875979 TI - Gianotti-Crosti syndrome related to rotavirus infection. PMID- 9875980 TI - Unilateral hyperhidrosis, callosities, and nail dystrophy in a boy with tethered spinal cord syndrome. PMID- 9875981 TI - Progeroid syndrome: association with connective tissue disease? PMID- 9875982 TI - Jacquet diaper dermatitis: a diagnosis of etiology. PMID- 9875983 TI - Education and child safety: a children's crusade. PMID- 9875984 TI - Medical mummies: the history of the Burns Collection. PMID- 9875985 TI - Survey of gross anatomy courses in the United States and Canada. PMID- 9875986 TI - The child as a projectile. AB - Unintentional injury is the leading cause of death in children under the age of fourteen. The majority of these injuries/deaths occur when the child becomes airborne during an accident. The most common mechanisms by which children become airborne are motor vehicle collisions, bicycling accidents, and falls. A head injury is seen in a significant number of children in this setting. This includes injury to the scalp, skull, coverings of the brain, or the brain itself. These injuries are the most common cause of death in children resulting from unintentional injury. Other typical injuries include external bruises and abrasions, extremity fractures, and bruising or bleeding of internal organs. We propose to name this constellation of injuries the projectile child syndrome. This refers to those injuries occurring in infants and children as a result of becoming airborne during the events of an accident. The pattern of injuries seen as related to the anatomy of the child is stressed. A review of the impact to society and guidelines for prevention are presented. PMID- 9875987 TI - Exploring brain circuitry with neurotropic viruses: new horizons in neuroanatomy. AB - There have been substantial advances in methods for defining connections among neurons over the past quarter century. However, most tracers have been limited in their ability to define populations of functionally related neurons that contribute to a multisynaptic circuit because they are not transported across synapses. As a result, the large body of literature that has employed these tracers has established regional associations between regions that must be further explored with electron microscopy and electrophysiological methods to define the synaptic relations among constituent neurons. Recently, neurotropic alpha herpesviruses have been used to visualize ensembles of neurons that contribute to polysynaptic networks. These pathogens invade permissive cells, replicate, and pass transynaptically to infect other neurons. In effect, the viruses become self-amplifying tracers whose natural tropism and invasiveness define populations of functionally related neurons. The recent increase in the use of this experimental approach has emerged from advances in our understanding of the life cycle of these viruses and the resulting evidence in support of specific transynaptic passage of progeny virus rather than infection by lytic release into the extracellular space. This article reviews the advances that have made this a viable experimental approach and considers ways in which this method has been creatively used to illuminate aspects of nervous system circuit organization that could not be defined with conventional tracers. PMID- 9875988 TI - Optometric educators: contributing to the future of optometry. PMID- 9875989 TI - Bilateral macular dysplasia in fragile X syndrome. AB - PURPOSE: Few studies have investigated the eye and vision dysfunctions of children with the fragile X syndrome. CASE REPORT: We report on a preschool boy with bilateral macular dysplasia and fragile X syndrome. His ocular features and phenotypic and genetic expressions are described. His mentally normal mother was identified as an expansion mutation carrier, and his older sister has learning disabilities, astigmatic refractive error, squint, and mild ptosis. Intrauterine infection has been excluded. CONCLUSION: To our knowledge, the association of macular dysplasia with fragile X syndrome has not been reported. The finding of macular dysplasia might be a coincidental developmental disorder and not a part of the syndrome. It could be considered a condition causing visual deficit with nystagmus in fragile X syndrome. PMID- 9875990 TI - The role of Pseudomonas aeruginosa biofilm in the attachment of Acanthamoeba to four types of hydrogel contact lens materials. AB - PURPOSE: The effect of the previous coating of a contact lens surface with Pseudomonas biofilm on adsorption of Acanthamoeba onto four types of hydrogel materials was investigated. METHODS: Hydrogel contact lens quarters from each of the four FDA groups of hydrogel materials were incubated for at least 12 h in a suspension of 10(7)/ml of Pseudomonas aeruginosa (ATCC 27853) to coat their surfaces with biofilm. After rinsing, the lenses were incubated for 90 min in 5 x 10(5)/ml of Acanthamoeba castellanii trophozoites. New, uncoated lens quarters were incubated in the Acanthamoeba suspension as controls. After rinsing, all adsorbed trophozoites on one surface of each lens quarter were counted by direct light microscopy. Adsorption was expressed as numbers of amoebae per square centimeter of lens surface, and nonparametric data analysis was performed. RESULTS: Acanthamoeba adsorption to new, uncoated lenses was greater for ionic materials (groups 3 and 4) than for the nonionic materials (groups 1 and 2). Pseudomonas biofilm increased adsorption on all four lens types. CONCLUSION: Pseudomonas biofilm enhanced adsorption of Acanthamoeba on all lens types studied, but the adsorption to nonionic materials was significantly less. This suggests that all lens wearers may be at increased risk for Acanthamoeba infection if lenses are previously contaminated with bacterial biofilm, but this risk may be reduced by the use of certain lens types (low water content, nonionic materials). PMID- 9875992 TI - Location and stability of a newly established eccentric retinal locus suitable for reading, achieved through training of patients with a dense central scotoma. AB - Six patients, median age 71 years, with a dense central scotoma in one eye and a median visual acuity of 0.06 (20/330) in the same eye, were all (100%) shown by means of fundus photography including a fixation target to preferably use an unfavorable retinal locus for fixation, i.e., within the lesion (scotoma). None of the patients was able to read novel text with the affected eye. A computer and video display system were used to determine the most suitable area above or below the visual field scotoma (below or above the retinal lesion) for reading and the magnification needed at this eccentricity. The same setup was also used for an introductory training in reading single words as well as scrolled text with the aim of establishing a preferred retinal locus (PRL) at a favorable, eccentric position, the trained retinal locus (TRL). Thereafter, the patients were provided with strong positive lenses (median power, 40 D) for reading printed text at a very short reading distance (median, 2.5 cm), first single words, above and below which help lines were printed to facilitate eccentric fixation, and finally, novel text. The total training time was 4 to 5 h. Thereafter, fundus photography showed that five of the patients (83%) used their TRL as their PRL. Reading speed was 71 words per minute (median). Our results seem to indicate that an eccentric PRL favorable for effective reading can be established through training and that a fairly low number of training sessions is required. PMID- 9875991 TI - Scotopic sensitivity: relation to age, dietary patterns, and smoking status. AB - PURPOSE: Although previous data suggest that rod-mediated sensitivity decreases with age, this decrease may be insignificant when only healthy individuals younger than 65 years are considered. In this study, we assess the relationship between age and scotopic sensitivity loss in subjects younger than 65 years to determine whether scotopic sensitivity losses can be detected when confounding factors are considered (including iris color, smoking status, and dietary patterns) and a large sample size is used. METHODS: A total of 121 subjects (aged 20 to 63 years) were tested under dark-adapted (scotopic) conditions. Scotopic sensitivity was measured as absolute thresholds to a 2.8 degree, 550-nm test presented at 6 degrees in the temporal hemiretina. Stimuli were presented in Maxwellian view. RESULTS: When all the subjects were considered together, there was a slight nonsignificant trend for scotopic sensitivity to decline with age (p < 0.11). This tendency was largely driven by the older (45 to 63 years) past and never smokers and was statistically significant (p < 0.024 and p < 0.05, respectively) when those two groups were analyzed separately. Scotopic sensitivity for the younger (20 to 44 years) past, current, and never smokers did not decline with age. When all the variables were considered in a general model, dietary intake of vitamin E explained a significant amount of the variation in scotopic sensitivity (p < 0.03). No relationships were found between scotopic sensitivity and iris color. CONCLUSIONS: Age-related losses in scotopic sensitivity before age 65 are slow. Moreover, individual variations in scotopic sensitivity for younger subjects is minimal, even in the presence of dramatic stressors such as long-term, heavy exposure to cigarette smoke. These data suggest that measurements of scotopic sensitivity may not be good indicators of the retinal health of individuals younger than 65 years. PMID- 9875993 TI - Mental effort required for walking: effects of retinitis pigmentosa. AB - PURPOSE: To determine whether retinitis pigmentosa (RP) increases the mental effort required for walking. METHODS: A dual-task methodology was used; walking speed and reaction time (RT) to randomly emitted tones were measured in 13 persons with RP and 29 persons with normal vision. Measures of RT were used to estimate the mental effort required for walking. In a second experiment, 15 persons with RP and 17 persons with normal vision navigated a "simple" and a "complex" route. RESULTS: The RP subjects had longer RT compared with the normal vision subjects when walking the complex but not the simple route. Normalized to each person's baseline, RT while walking the complex route was significantly correlated with log contrast sensitivity (r = -0.63) and log retinal area (r = 0.64) in the RP group. The amount of mental effort required for walking was shown to also depend on familiarity with the route. CONCLUSIONS: Route complexity is critical in determining whether walking requires more mental effort for persons with RP than for persons with normal vision. The magnitude of mental effort required for mobility covaries with the visual impairment measures which correlate with mobility performance in RP. PMID- 9875994 TI - Repeatability of refraction and corrected visual acuity in keratoconus. The CLEK Study Group. Collaborative Longitudinal Evaluation of Keratoconus. AB - PURPOSE: The purpose of the test-retest phase of the Collaborative Longitudinal Evaluation of Keratoconus (CLEK) Study was to determine the repeatability of the various parts of the CLEK Study protocol. This paper presents the test-retest parameters of the refraction protocol. METHODS: We examined 138 CLEK Study eligible patients on two occasions (median, 90 days; range, 22 to 268 days). All patients underwent subjective refraction on two occasions, and contact lens over refractions were performed either over the patient's habitual rigid contact lenses or over a trial rigid contact lens equal in base curve to the steep keratometric reading in nonrigid contact lens wearers. RESULTS: Mean interoccasion differences +/- SD were -0.32 +/- 2.91 D and -0.17 +/- 1.39 D for subjective refraction sphere and cylinder power, respectively, and the mean absolute difference for subjective refraction cylinder axis was 18.1 +/- 20.2 degrees. The mean interoccasion difference +/- SD for high-contrast visual acuity with subjective refraction was 0.38 +/- 10.9 letters correct. Mean interoccasion differences +/- SD were -0.11 +/- 0.81 D and 0.02 +/- 0.67 D for contact lens over-refraction sphere and cylinder power, respectively, and the mean absolute difference for contact lens over-refraction cylinder axis was 11.6 +/- 9.9 degrees. The mean interoccasion difference +/- SD for visual acuity with contact lens over-refraction was 0.50 +/- 5.2 letters correct and 0.71 +/- 6.9 letters correct for high- and low-contrast visual acuity, respectively. CONCLUSIONS: The repeatability of subjective refraction in keratoconus patients is good but somewhat lower than that found in nondiseased eyes. Only 36% of our repeat measures of sphere power from subjective refraction fell within 0.50 D of each other, compared with more than 90% in studies of normal eyes. PMID- 9875995 TI - The magnitude and distribution of open-loop accommodation using three different methods of opening the loop. AB - PURPOSE: The present study examines the magnitude, distribution, and relationship of open-loop accommodation obtained using the three most common methods of opening the accommodation loop. METHODS: Open-loop accommodation was measured in 93 young, emmetropic subjects using a Canon R1 objective infrared optometer, and the accommodation loop was opened using the following methods: (1) dark empty field (DA), (2) bright empty field (BA), and (3) viewing a target through a small artificial pupil (PA). RESULTS: PA was found to elicit significantly (p = 0.0001) higher values of open-loop accommodation than either DA or BA and demonstrated a much wider distribution of values than DA or BA. A further experiment demonstrated that the higher PA was attributable to the proximal effect of placing a small artificial pupil close to the eye. CONCLUSIONS: Our results suggest that using a small artificial pupil to open the accommodation loop may not produce a veridical measure of open-loop accommodation. PMID- 9875996 TI - Dark focus values measured by retinoscopy: a clinical chart review. AB - The present study investigated the utility of dark retinoscopy as a technique to measure the dark focus of accommodation during routine clinical examinations. Using data from clinical examinations collected during a period of 6 months, a clinical chart review quantified dark focus and mean spherical equivalent refractive error for 480 patients ranging in age from 6 to 55 years. Results showed significant variations in mean dark focus values over the age groups. All groups showed wide interpatient differences, with mean dark focus values that were significantly nearer than the farpoint. Individual dark focus values ranged from 2.25 to -1.00 D (hyperopic), with an overall mean of 0.53 D. Potential clinical applications of dark focus-based correction are discussed. PMID- 9875997 TI - Virtual reality flexible sigmoidoscopy simulator training: impact on resident performance. AB - BACKGROUND: Flexible sigmoidoscopy, a core skill for the primary care physician, requires learned hand-eye skills that can be difficult to master during residency training. With recent advances in virtual reality simulation technology, simulated flexible sigmoidoscopes are available to family medicine residents for training before their initial and subsequent live patient examinations. The purpose of the study was to determine whether a virtual reality flexible sigmoidoscope simulator would improve the hand-eye skills and various performance parameters in a live patient. METHODS: Residents were assigned to a control (n = 5) or experimental group (n = 5) in which the experimental group trained on a virtual reality sigmoidoscopy simulator before their first sigmoidoscopies on live patient volunteers. After the initial live patient sigmoidoscopies, both control and experimental groups trained on the simulator so that it was possible to evaluate presimulator and postsimulator training effects on live patient performance and to compare speed and skill between the groups at different levels of training. RESULTS: Training on the virtual reality simulator produced substantial improvements in examination times and hand-eye skill measures. After 6 to 10 hours of training on the simulator, the experimental group achieved significantly faster insertion times to 30 cm (119 versus 357 sec, P = 0.03), 40 cm (211 versus 518 sec, P = 0.03), and a shorter mean length of examination (323 versus 654 sec, P = 0.01). There was also significant improvement of hand-eye skill measures of the experimental group in directional errors (1.6 versus. 8.6, P < 0.01), percentage of colon visualized (79 versus 45 percent, P = 0.02), and viewing quality of examination when compared with the control group's initial performance on live patients. Resident survey findings after the study confirmed the trainee's perception of the benefit of the simulator training. CONCLUSIONS: This study shows the value of virtual reality simulator training for accelerating the development of the hand-eye skills to perform adequate sigmoidoscopy. PMID- 9875998 TI - Oral health care issues in HIV disease: developing a core curriculum for primary care physicians. AB - BACKGROUND: Given the high occurrence of oral manifestations in patients infected with human immunodeficiency virus (HIV), the relative ease in recognizing these manifestations on physical examination, and their potential impact on the health care and quality of life in these patients, it is critical to provide adequate training for primary care physicians in this area. METHODS: Based on a review of the published literature and the consensus of a national panel of primary care physicians and dentists with clinical and research expertise in this area, a core curriculum was developed for primary care physicians regarding oral health care issues in HIV disease. RESULTS AND CONCLUSIONS: We describe the process of developing the core curriculum of knowledge, skills, and attitudes regarding oral health care issues in HIV disease. The final curriculum is in a format that allows for easy accessibility and is organized in a manner that is clinically relevant for primary care physicians. PMID- 9875999 TI - Clinical feasibility of a free-weight strength-training program for older adults. AB - BACKGROUND: An emerging trend in the field of gerontology is the recognition that older adults, especially the frail elderly population, can increase their levels of strength and thus improve functional capability. Social acceptance of physical frailty and provision of care and assistance to dependent persons has now turned to helping the frail elderly adults maintain or improve functional independence. METHODS: The purpose of our study was to show the feasibility and effectiveness of a low-cost strength-training program using free weights for increasing strength and functional fitness among older adult volunteers. Participants aged 73 to 94 years were residents of a multilevel care retirement community in Columbia, SC. The strength-training program, led by an instructor, used dumbbells and ankle weights and was conducted in a multipurpose recreation room at the retirement facility. RESULTS: Functional performance measures (timed chair stand, 6-meter walk, stair climb, balance) handgrip strength, and self-assessment of activity level were outcome measures. All 25 participants completed the strength training program. The average program adherence rate of all participants was 87 percent. No participant injuries or other adverse effects were observed. Functional performance measures improved significantly among program participants, with the greatest improvement in the timed chair stand (33.5 percent improvement) and the stair climb (17.6 percent improvement). CONCLUSION: Free-weight strength-training programs are appropriate for older adults, can be implemented in community settings, and are associated with significant improvement in functional performance. PMID- 9876000 TI - Use of the health history as a psychiatric screening tool. AB - BACKGROUND: Numerous studies indicate that most mental health services are provided in the primary care medical setting and that problems exist with misdiagnoses and lack of recognition of mental health disorders. The purpose of our investigation was to determine whether patient responses on a typical medical health history form could be used to predict depression or anxiety. METHODS: New adult patients at a university community family practice clinic were surveyed during a 6-month period. Study patients completed a health history form and standardized inventories of anxiety and depression. RESULTS: The study sample included 187 patients. Positive responses to mood-related symptoms reported on the health history best predicted anxiety and depression. Nonmood symptoms were also significant, although not as powerful, predictors of these disorders. A significant relation existed between total number of positive symptoms and psychiatric diagnoses, which continued when mood symptoms were removed from the analyses. Results were used to develop physical, nonmood primary care symptom profiles that could be used to screen for anxiety and depression. CONCLUSION: Providing physicians the means to improve mental health diagnostics can help advance patient care and health care system outcomes. PMID- 9876002 TI - Using outcomes to improve quality of research and quality of care. PMID- 9876001 TI - Tuberculous cerebellar abscess. AB - BACKGROUND: Tuberculosis is an important illness with an increasing occurrence. Although considered primarily a pulmonary disease, tuberculosis can affect any organ system. Central nervous system involvement is potentially devastating and occurs with escalating frequency in both immunocompetent and immunologically incompetent populations. METHODS: This case report involved a patient admitted to the authors' inpatient service. Data were obtained from the patient's medical record. MEDLINE and Index Medicus literature searches were conducted for the years 1977 to the present, with cross-references for earlier articles. RESULTS AND CONCLUSIONS: A 36-year-old previously healthy, immunocompetent woman with a diagnosis of extrapulmonary tuberculosis and no determined primary focus was found to have a cerebellar abscess. Treatment included surgical excision of the abscess combined with 6 months of antituberculosis therapy. Diagnostic tools included the tuberculosis skin test, smears and culture of specimens, computed tomographic scans, and rapid assays based on nucleic acid amplification, ie, polymerase chain reaction. The polymerase chain reaction has great potential for rapid diagnosis of Mycobacterium tuberculosis, particularly when there might be few bacilli, as in pleural, peritoneal, or cerebrospinal fluid. Appropriate therapy for tuberculous cerebellar abscess includes standard antituberculosis medications for 6 to 9 months and surgical excision of the abscess. PMID- 9876003 TI - Recognition and management of oral health problems in older adults by physicians: a pilot study. PMID- 9876004 TI - Transient hyperthyroidism in a patient taking dietary supplements containing kelp. PMID- 9876005 TI - Unintentional carbon monoxide poisoning from an unlikely source. PMID- 9876006 TI - Legacy of the secret war: medical needs in the UXO-contaminated areas in Laos. PMID- 9876007 TI - Border crossings: on the boundary of the physician-patient relationship. PMID- 9876008 TI - Will virtual reality simulators end the credentialing arms race in gastrointestinal endoscopy or the need for family physician faculty with endoscopic skills? PMID- 9876009 TI - Irritant-induced anosmia. PMID- 9876011 TI - Modulation of ras p21 oncoprotein levels and DNA strand breakage in human cells with chemotherapeutic agents and/or deferoxamine. AB - Oncogenes are involved with the regulation of cellular proliferation. Ras oncogenes can be activated by chemical treatment and any increased activity could be modulated by further chemical treatment. In the present study, therefore, ras p21 protein expression was examined in in vitro cultures of human lymphocytes treated with mitomycin C and in the human colon adenocarcinoma Caco-2 cell line treated with doxorubicin with and without deferoxamine. Both chemotherapeutic agents act partially through oxygen radical mechanisms. Increases in p21 protein levels were seen with mitomycin C but no clear response was seen with doxorubicin. However, deferoxamine, with and without doxorubicin, altered p21 expression. Deferoxamine is an iron chelator so these results support the hypothesis that oxygen radicals were responsible for the altered p21 protein levels. Modulating responses were confirmed by measuring DNA strand-breakage in the Comet assay after treatment with doxorubicin and deferoxamine. Alterations of ras p21 protein expression in vitro might prove a suitable system for examining modulating effects on chemical carcinogens. PMID- 9876010 TI - Lack of UDS activity in the livers of rats exposed to allylisothiocyanate. AB - Allylisothiocyanate (AITC) has been evaluated for its ability to initiate unscheduled DNA synthesis (UDS) in the livers of male rats in vivo. Specific Pathogen Free outbred albino Hsd/Ola Sprague-Dawley rats were exposed by oral gavage to 37.5 or 125 mg/kg AITC in corn oil and hepatocytes assessed for UDS by autoradiography 2 and 14 h later. AITC did not induce UDS at either dose level at either time point. These data are consistent with all other evidence indicating that AITC does not act as a genotoxin in vivo, despite positive response in some in vitro screening assays. The reported occurrence of benign bladder papillomas in male rats but not female rats or mice of either sex is consistent with non genotoxic action and may be attributed to chronic irritation of the bladder epithelium by AITC and its cysteine conjugate metabolite excreted by male rats in unusually concentrated form. It is concluded that the weight of evidence is insufficient to regard AITC as a genotoxin capable of human carcinogenicity. PMID- 9876012 TI - Developmental toxicity of indium chloride by intravenous or oral administration in rats. AB - Pregnant rats were treated with a single intravenous or oral administration of indium chloride (InCl3) on day 9 of pregnancy and their fetuses were examined for growth and malformation on day 20 of pregnancy. By intravenous administration, fetal weight was significantly decreased and the incidences of fetal mortality and malformation were significantly increased at 0.4 mg In/kg. Fetal malformations of the tail and digits, e.g., kinked tail, brachyury, and oligodactyly, were observed at high incidences. By oral administration, similar tendencies in the fetal effects were observed, but there were no significant differences compared to the control even at 300 mg In/kg. Indium concentrations in the serum of pregnant rats showed low bioavailability of indium by oral administration. It was concluded from these results that indium showed teratogenicity in rats. Oral treatment with indium may be developmentally toxic at 300 mg In/kg, but this is difficult to state with certainty given the limited number of animals that were used in this study. PMID- 9876013 TI - Genotoxic activity of 4,4',5'-trimethylazapsoralen on plasmid DNA. AB - The genotoxic activities of 8-methoxypsoralen (8-MOP) and 4,4',5' trimethylazapsoralen (4,4',5'-TMAP) on plasmid DNA have been compared. In a previous work, 4,4',5'-TMAP, a methyl derivative of a psoralen isoster, had shown potential photochemotherapeutic activity. The mutagenic activity of mono- and bifunctional lesions caused by these compounds was evaluated both after UVA irradiation, which causes the formation of both kinds of lesions, and after a two step irradiation procedure of the psoralen-plasmid DNA complex, which allowed monoadducts and interstrand crosslinks to be studied separately. Furthermore, we used a procedure that allowed us to evaluate both the mutagenic and recombinogenic activity of the two compounds. Results indicate that the most important difference between 8-MOP and 4,4',5'-TMAP consists in their mode of photoreaction with DNA rather than in their mutagenic potential. In fact, in all of the experimental procedures, 4,4',5'-TMAP shows a lower ability than 8-MOP to generate interstrand crosslinks. However, when comparable toxicity levels are reached, the two compounds show the same mutagenic potentiality. PMID- 9876014 TI - Aflatoxin B1-induced micronuclei and cell cycle alterations in lung and bone marrow cells and their modulation by Piper argyrophyllum extract. AB - Aflatoxin B, (AFB1) is a clastogen that causes cellular damage by covalent modification of nucleic acids. In this investigation, male rats were injected i.p. with AFB1 (8 mg/kg b.w.) in DMSO and the same dose of AFB1 was also administered intratracheally (i.t.) to the animals separately. The animals were killed after 26 h of the carcinogen treatment, femur bone was removed, and bone marrow cells were isolated and stained with Mayer's hematoxylin and eosin. Micronuclei (Mn) were scored by using light microscopy. Bronchoalveolar lavage (BAL) was prepared from rats administered AFB1 i.t. A part of BAL was fixed with 70% ethanol, stained with the fluorochrome DAPI, and analysed for cell cycle variations; the other part of the lavage was used for making slides to record Mn with a fluorescent microscope. A significantly greater proportion of lung cells were found to enter cell cycle with extended S-phase due to AFB1 treatment. Mn were induced in polychromatic erythrocytes (PCE) as compared to normochromatic erythrocytes (NCE) in the bone marrow of AFB1-treated rats, where there was nearly a three-fold increase in the number of Mn of bone marrow cells. The administration of AFB1 resulted in a two-fold rise in the Mn in the lung cells. The effect of BSO, DEM, and PB, the modulators of AFB1 metabolism, was studied on AFB1-induced Mn formation. A significant increase in the Mn score in PCEs of BSO- and DEM-treated rats was noted, while a slight reduction in the Mn score was noted in the case of PB-treated rats. The administration of the methanol extract of the leaves of Piper argyrophyllum (taken up in DMSO) to rats for a week exhibited normalising effect on AFB1-induced Mn in bone marrow cells. These observations record the induction of Mn in lung cells due to AFB1 for the first time. We propose the utility of AFB1-induced Mn as a model for screening plant extracts as inhibitors of genotoxicity. Prevention of genotoxic changes described above by phytochemicals is being pursued in our Laboratories. PMID- 9876015 TI - The role of transcriptional control during spermatogenesis. PMID- 9876016 TI - The use of two fluorescent dyes to identify sperm in a competitive binding assay to oocytes. AB - The relationship of most sperm laboratory assays to male fertility is inconsistent. Assays that measure traits required to fertilize oocytes are expected to have the most predictive value. A new assay that measures the competitive ability of two sperm samples to bind to oocytes was developed. Two populations of sperm were labeled using a pair of lipophilic dyes. A concentration of 75 microM of the two dyes, DiQ (4-[4-(dihexadecylamino)styryl]-N methylquinolinium iodide; an orange-red dye) and DiOC16 (3,3' dihexadecyloxacarbocyanine perchlorate; a yellow-green dye), intensely stained 66 and 73% of sperm, respectively, without affecting sperm motility or oocyte binding ability. Because sperm could be stained with fluorescent dyes, sperm from two semen samples were mixed together in a droplet, and oocytes were added to allow sperm to bind oocytes competitively. Oocyte-bound sperm from each sample were counted. Binding was specific; nonspecific sperm binding was estimated by sperm bound to two-cell mouse embryos and averaged one to three sperm per embryo. Staining with DiQ or DiOC16 did not affect oocyte-binding ability since more than 80% of the sperm bound were stained with either dye. Furthermore, if different ratios of DiQ- or DiOC16-stained sperm from the same ejaculate were prepared in droplets and oocytes were added, the percentage of sperm bound to the oocytes reflected the percentage of sperm in the droplet; there was no differential effect of either dye. This assay used fixed oocytes because sperm bound equally to fixed and fresh bovine oocytes. This competitive oocyte-binding assay allows one to make a series of pairwise comparisons between a group of males or to include an internal control sample in sperm-oocyte binding assays. This assay may allow more accurate prediction of the oocyte-binding ability of sperm. PMID- 9876017 TI - Expression of cyclin-dependent kinase 5 and associated cyclins in Leydig and Sertoli cells of the testis. AB - In this study, we examined the expression and subcellular localization of cyclin dependent kinase 5 (Cdk5), cyclin D1, and cyclin E in Leydig and Sertoli cell lines that were cultured with 7.5, 1.0, 0.5, or 0% serum (mixture of a 2:1 ratio of horse serum and fetal bovine serum) and in the developing rat testis to verify the possible functions of Cdk5, cyclin D1, and cyclin E in the testis. The abundance of Cdk5 and cyclin E in the Leydig cell line, TM3, was significantly reduced at low serum concentrations. In contrast, serum concentration had no effect on Cdk5 and cyclin E levels in the Sertoli cell line, TM4. Cyclin D1 was detected by western blot analysis in TM4 cells only, and its abundance was serum dose dependent. The kinase activity of Cdk5 in TM3 and TM4 cells that were cultured at various serum concentrations coincided with the levels of Cdk5 expression. Immunohistochemical staining for Cdk5 and cyclin E revealed nuclear and cytoplasmic distribution, both in TM3 and TM4 cells. Moreover, cyclin D1 immunoreactivity was only detected in TM4 cells. In the developing rat testis, Cdk5 expression was most prominent at 2 and 3 weeks after birth. Cyclin D1 was strongly expressed at 1 and 2 weeks in premature rat testes. On the other hand, cyclin E was highly expressed in the adult testis. Immunohistochemical localization of Cdk5, cyclin D1, and cyclin E in 1-week-old and adult rat testes revealed expression in both Leydig and Sertoli cells. Our results suggest that Cdk5 in TM3 and Leydig cells of the testis might play a role in cell cycle regulation, whereas Cdk5 in TM4 and Sertoli cells of the adult testis might have some additional functions besides control of proliferation. PMID- 9876018 TI - Sperm motility enhancement by nitric oxide produced by the oocytes of fathead minnows, Pimephelas promelas. AB - The effects of nitric oxide (NO) on sperm motility were examined in the fathead minnow, Pimephelas promelas, using computer-assisted sperm analysis (CASA). The observed effects underscore the dual nature of NO as both a low-concentration regulatory agent and, at higher doses, a cytotoxic agent. At 1 x 10(-6) M concentration, NO donor sodium nitroprusside (SNP) enhanced sperm motility percentages and increased CASA velocity parameters curvilinear velocity, straight line velocity, and average path velocity, whereas 1 x 10(-2) M concentration inhibited percent motility and decreased velocities. Fathead minnow ova-produced NO was subsequently trapped as a paramagnetic ferrous iron complex and detected by electron spin resonance spectroscopy. The distinctive triplet spectrum, with a(N) = 12.5G and g(iso) = 2.04, was recorded during a critical 5-minute period following laying. Nitric oxide synthase (NOS) was histochemically localized at the micropyle of mature unfertilized fathead eggs, and an inhibitor of NOS blocked histochemical staining. CASA analysis of sperm motility in the presence of ovaproduced NO over an 8-minute time course reveals an optimum motility enhancement at 4 minutes that is similar to the effect of 1 x 10(-6) M SNP. This transient NO production by freshly laid ova and the localization of NOS near the site of sperm entry, together with the motility-enhancing effect of 1 x 10(-6) M SNP on sperm, indicates an active role for low-concentration NO in fertilization. PMID- 9876019 TI - Regulation of sperm function by protein tyrosine phosphorylation in diverse wild felid species. AB - Protein tyrosine phosphorylation is associated with sperm capacitation and the acrosome reaction in several mammalian species. Changes in phosphorylation of a 95-kDa protein in human, mouse, and domestic cat spermatozoa are known to be influenced by capacitation and exposure to zona pellucida (ZP) proteins. We previously reported diminished phosphorylation of 95- and 160-kDa proteins in spermatozoa from teratospermic cats, compared with normospermic domestic cats. To determine if these proteins and mechanisms are present in other species in the phenotypically diverse Felidae family, we examined the relationship between tyrosine-phosphorylated sperm proteins and sperm morphology in the leopard cat (approximately 65% normal sperm/ejaculate), tiger (approximately 65%), clouded leopard (approximately 15%), and cheetah (approximately 30%). Furthermore, we investigated the involvement of cyclic adenosine monophosphate (cAMP) in the regulation of sperm protein tyrosine phosphorylation. Specifically, we assessed the following: 1) presence of tyrosine-phosphorylated proteins in sperm extracts; 2) changes in protein tyrosine phosphorylation after sperm capacitation and swim up separation; 3) impact of tyrosine kinase inhibition on leopard cat sperm protein phosphorylation and ZP penetration; and 4) involvement of a cAMP dependent pathway in the regulation of protein tyrosine phosphorylation. Immunoblotting analysis with anti-phosphotyrosine antibody (PY20) indicated that a 95-kDa protein was present in all four species. Additional phosphorylated proteins were detected in the leopard cat (145- and 175-kDa proteins), tiger (185 kDa protein), clouded leopard (160- and 190-kDa proteins), and cheetah (115- and 155-kDa proteins). Sperm capacitation in vitro increased phosphorylation of one or more proteins in the leopard cat, tiger and clouded leopard, but not in the cheetah. Although swim-up separation increased the proportion of morphologically normal spermatozoa in the clouded leopard and cheetah, no changes were observed in phosphorylation of the 95-kDa sperm protein. Thus, phosphorylation of the 95 kDa protein appeared to be related to the condition of teratospermia. Exposing leopard cat spermatozoa to the tyrosine kinase inhibitor, tyrphostin, reduced (P < 0.05) phosphorylation of the 95- and 145-kDa proteins, as well as ZP penetration, without affecting sperm motility. Similarly, when spermatozoa were incubated in the presence of cAMP analogs or active and inactive stereoisomers of cAMP, phosphorylation of sperm proteins was either stimulated or inhibited. Together, these data suggest that protein tyrosine kinase mechanisms appear conserved within the family Felidae and are regulated by a cAMP/protein kinase A pathway. PMID- 9876020 TI - Evidence for cross-talk between Sertoli and germ cells using selected cathepsins as markers. AB - To examine whether proteases are possibly involved in cellular migration and/or spermiation when developing germ cells translocate across the seminiferous epithelium during spermatogenesis, in situ hybridization was used to localize messenger RNA (mRNA) transcripts of cathepsin L, D, and S in the epithelium at different stages of the spermatogenic cycle in the rat. Cathepsin L mRNA was found to localize almost exclusively near the basal lamina of the epithelium. At stages VI and VII of the cycle before spermiation, the signal of cathepsin L mRNA was so intense that it formed a complete dark precipitate near the basal lamina encircling the entire tubule. At stage VIII, the expression of cathepsin L was completely abolished, and no staining of cathepsin L mRNA was seen in the epithelium. The mRNA of cathepsin D and S was found near the basal lamina, a finding consistent with their localization in Sertoli cells and possibly primary spermatocytes in almost all stages, but peaked at stages VII-IX and VII-VIII of the cycle, respectively, at the time before and during spermiation. These results illustrate the possible involvement of these proteases in facilitating germ cell movement and spermiation. To examine whether germ cells express any of these cathepsin genes, spermatocytes with a purity of greater than 95% were isolated from 15-day-old rat testes by Percoll gradient centrifugation for reverse transcriptase-polymerase chain reaction. It was found that primary spermatocytes expressed multiple cathepsin genes, including cathepsin B, C, D, H, L, and S. Furthermore, the expression of cathepsin L by germ cells isolated from 15-day-old rats (largely spermatocytes and spermatogonia) can be stimulated by Sertoli cell enriched culture medium in a dose-dependent manner, but not by germ cell conditioned medium. These results reveal that germ cell function can be regulated by Sertoli cells. Moreover, these results suggest that germ cells may play an active role in the overall testicular protease expression. Also, we present evidence suggesting there is cross-talk between Sertoli and germ cells, since the expression of cathepsin L in each cell type is regulated by one another via either soluble factors or cell-cell contact. PMID- 9876021 TI - Effects of cryopreservation on bull sperm head morphometry. AB - Artificial insemination using cryopreserved semen is a common management tool of the contemporary livestock producer. However, cryopreservation is detrimental to sperm function and fertility, killing some 50% of the spermatozoa during the process. Prediction of cryopreservation damage from prefreeze samples remains elusive. Computer-automated sperm head morphometry was used in this study to determine the effects of cryopreservation on bovine sperm head morphometry. Semen was collected from 18 bulls and was divided. One portion was extended to 200 x 10(6) sperm/ml and a microscope slide was prepared, while the remaining portion was cryopreserved in a Triscitrate-yolk extender. After thawing, the cryopreserved samples were prepared on microscope slides. All slides were air dried and were stained with hematoxylin and rose bengal. The morphometric dimensions for length, width, width/length, area, and perimeter for a minimum of 200 sperm heads were analyzed from each slide by computer-aided sperm head morphometry analysis, and the mean measurements were recorded. Bull sperm heads were significantly (P < 0.01) smaller in cryopreserved spermatozoa than in the companion extended samples for length (8.56+/-0.07 vs. 8.63+/-0.08 microm), width (4.39+/-0.05 vs. 4.48+/-0.05 microm), area (28.42+/-0.07 vs. 29.14+/-0.08 microm), and perimeter (23.33+/-0.21 vs. 23.70+/-0.23 microm) for all bulls. Width/length was also different (0.513 vs. 0.519). In addition, differences (P < 0.05) were found within 14 of 18 bulls for at least four of the morphometric parameters. The percent change in measures after cryopreservation were correlated (P < or = 0.05) to the variability of the extended sample. Variations in sperm head measurements were lower (P < or = 0.05) in extended samples of the four bulls in which no changes occurred than in extended samples of the remaining 14 bulls. These data suggest that the variability in sperm head measurements of individual bulls, or ejaculates, may be an indicator of sperm cryosurvivability. PMID- 9876022 TI - Quantification of apoptotic testicular germ cells in normal and methoxyacetic acid-treated mice as determined by flow cytometry. AB - Several studies have reported the occurrence and significance of programmed cell death (apoptosis) of testicular germ cells in mammals. In those studies, apoptotic germ cells were identified by morphological criteria or by in situ end labeling (TUNEL) and were enumerated from histological sections by semi quantitative and time-consuming techniques. In the present study, we have established a flow cytometric technique for quantification of TUNEL-positive cells in the mouse testis. Groups of five adult mice each received 0, 650, or 1300 mg/kg (IP) of methoxyacetic acid (MAA), and testes were collected 24 hours later. MAA is known to induce germ cell apoptosis in rodent testes. MAA induced a significant (P < 0.01) dose-dependent decline in the percentage of pachytene spermatocytes (4C cells). DNA strand breaks generated by the activation of endogenous endonuclease in the apoptotic germ cells were detected by the in situ labeling of the 3'-OH termini with biotinylated dUTP in the presence of terminal deoxynucleotidyl transferase (TUNEL technique). Histologically, TUNEL-positive germ cells were observed in control testes, and the number of these cells was visibly increased following MAA exposure. As determined by flow cytometry, four cell populations contained TUNEL-positive cells: 1C cells (round spermatids), 2C cells (mainly spermatogonia), S-ph cells (spermatogonial cells and preleptotene spermatocytes synthesizing DNA [the S-phase]), and 4C cells (primary spermatocytes). Analysis of the percentages of TUNEL-positive cells within each population yielded values of 1.57+/-0.23% for 1C cells, 1.65+/-0.27% for 2C cells, 6.26+/-1.03% for S-ph cells, and 3.24+/-0.39% for 4C cells. Hence, a substantial proportion of proliferating cells are undergoing apoptosis during normal spermatogenesis. The overall incidence of apoptotic cells among all testicular cells was around 2%. At 650 mg per kilogram of body weight, MAA induced a fourfold to eightfold increase (P < 0.001) in the percentage of TUNEL positive cells, compared with saline-treated controls, and, overall, 17% of testicular cells were apoptotic. This effect of MAA was most pronounced for S-ph and 4C cells, with 25-30% of cells being affected in each of those populations. At 1300 mg per kilogram of body weight, MAA had no further effect. These quantitative data demonstrate that 1) in the normal testis, it is mainly proliferating cells that undergo apoptosis, and 2) MAA induces primary spermatocyte loss by germ cell apoptosis. PMID- 9876023 TI - The primary culture of rat prostate basal cells. AB - The objective of this study was to identify the cells from rat prostate epithelium that attach and proliferate in primary culture. Minced ventral prostate was dissociated by DNAse/collagenase digestion, suspended in RPMI-1640 containing 10% fetal bovine serum, and subjected to Percoll centrifugation to separate the epithelial cells from stromal cells. With the use of lectins, it became possible to identify and monitor the fate of the dissociated epithelial cells held in suspension at 37 degrees C for several hours. In tissue sections of the rat prostate, Griffonia simplicifolia I-isolectin B4 (GSI-B4) specifically bound to basal cells, while Glycine maximus (soybean agglutinin [SBA]) was specific for secretory cells. Double staining with lectins and propidium iodide of dissociated cells revealed a preponderance of GSI-B4-positive live cells. The cells were plated in WAJC-404 medium supplemented with various factors, including insulin (5 ng/ml), transferrin (5 ng/ml), EGF (10 ng/ml), and bovine pituitary extract (30 microg/ml). Epithelial colonies that formed and proliferated from these cells also stained positively for GSI-B4 marker and for cytokeratins specific for basal cells as assessed by immunocytochemical staining. Proliferation was greater in cells grown on a collagen Type I matrix. These findings suggest that the epithelial cells that survived in suspension and proliferated in culture originated from basal cells of the rat prostate epithelium. PMID- 9876024 TI - Follicle-stimulating hormone is required for the initial phase of spermatogenic restoration in adult rats following gonadotropin suppression. AB - The role of follicle-stimulating hormone (FSH) in adult rat spermatogenesis is unclear. Although exogenous testosterone (T) restores spermatogenesis following gonadotropin-releasing hormone (GnRH) immunization or T plus estradiol (TE) treatments, an assessment of the independent action of T and FSH was not possible, as exogenous T treatment maintains serum FSH levels. We have used passive immunization against FSH to determine whether T alone is capable of reinitiating spermatogenesis after chronic and acute FSH withdrawal. Adult rats received T-filled Silastic implants 6 cm (T6) or 8 cm (T24) in length for 7 days in combination with either a polyclonal sheep antisera raised against rat FSH (FSHAb, 2 mg/kg SC daily) or control sheep immunoglobulin (ConAb) after either GnRH immunization (12 weeks) or TE treatment (9 weeks). The neutralizing capacity of the FSHAb was determined using a FSH in vitro bioassay; this analysis demonstrated that administration of FSHAb in vivo reduced FSH levels by >90%. Testes were fixed and germ cell number per testis quantified using the optical dissector. GnRH immunization reduced spermatogonia, pachytene spermatocytes, and round spermatids to 50, 13, and <1% of normal, respectively. T6 and T24 Silastic implants with the inclusion of the FSHAb did not increase the number of spermatogonia, pachytene spermatocytes, and round spermatids (50, 15, and 1% of normal, respectively). T6+ConAb treatment increased spermatogonial, pachytene spermatocyte, and round spermatid numbers to 74, 30, and 3% of normal, respectively (P < 0.05). No further increases were seen with T24 implants. TE treatment suppressed pachytene spermatocytes and round spermatids to 33 and 1% of normal, respectively (P < 0.05). T6+FSHAb treatment did not increase the number of pachytene spermatocytes and round spermatids (36 and 8%, respectively), whereas T6+ConAb treatment increased pachytene spermatocyte and round spermatid number to 50 and 28% of normal, respectively (P < 0.05). T24+FSHAb treatment increased the number of pachytene spermatocyte and round spermatids (56 and 22% of normal, respectively; P < 0.05), whereas T24+ConAb treatment increased these cells forms to 79 and 31% of normal, respectively. In conclusion, T alone is unable to restore spermatogenic cell populations in the setting of chronic FSH withdrawal. Although acute FSH withdrawal markedly impairs the restoration process, higher doses of T can partially compensate for the lack of FSH. These data suggest that FSH is important for the initial phase of spermatogenic restoration. PMID- 9876025 TI - Extender components and surfactants affect boar sperm function and membrane behavior during cryopreservation. AB - To determine how the individual components of extenders affected boar sperm function and membrane structure and to test a new surfactant's cryoprotective ability, boar sperm were cryopreserved in straws in BF5 extender plus or minus egg yolk plus or minus glycerol plus or minus a surfactant (Orvus ES Paste [OEP] or various concentrations of Pluronic F-127). After thawing, sperm function and fluidity of the isolated head plasma membrane (HPM) were determined. Total motility and adenosine triphosphate content (a measure of viability) were superior postthaw in sperm extended in egg yolk plus glycerol (P < 0.05); neither surfactant improved function. Egg yolk plus any other ingredients improved normal acrosome morphology, whereas a combined measure of motility and normal acrosome morphology was better in the presence of 0.33% OEP or 0.1% Pluronic F-127 (P < 0.05 vs. controls). Head plasma membrane was isolated from freshly collected spermatozoa and spermatozoa cryopreserved in the various extenders. Membrane fluidity was monitored with the probes cis-parinaric acid (cPNA), transparinaric acid (tPNA), and 1,6-diphenyl-1 ,3,5-hexatriene (DPH). The cPNA and the DPH monitor the fluidity of gel and liquid-crystalline areas of the membrane, whereas the tPNA preferentially monitors the gel-phase domains of the membrane. Additionally, DPH monitors the hydrophobic core of the bilayer. In the HPM from fresh sperm, the fluidity of each domain changed over time in a manner unique to that domain, and the behavior of the DPH domain varied among boars. The fluidity dynamics of each domain responded uniquely to cryopreservation. The cPNA domain was unaffected, the tPNA domain was altered by four of the eight extenders, and all extenders affected the fluidity of the DPH domain. Membrane structure was significantly correlated with cell function for sperm cryopreserved in extenders that preserved viability and motility. Sperm cryopreserved in egg yolk plus glycerol plus either OEP or 0.1% Pluronic F-127 functioned best when the bulk domains were less fluid initially and the gel domain solidified more slowly. Therefore, the behavior of domains in the HPM of boar spermatozoa is affected by cryopreservation and is related to the postthaw function of boar sperm cryopreserved in different extenders. PMID- 9876026 TI - [Tyr1]-nociceptin and nociceptin have similar naloxone-insensitive erectile activity in the cat. AB - The heptadecapeptide nociceptin, also known as Orphanin FQ, is a newly discovered endogenous ligand for the opioid-like G-protein-coupled receptor ORL1. The present study was undertaken to investigate responses to intracavernosal injections of the nociceptin analog [Tyr1]-nociceptin and to investigate the effects of naloxone on erectile responses in anesthetized cats to [Tyr1] nociceptin and to nociceptin. Intracavernosal injections of [Tyr1]-nociceptin and of nociceptin in doses of 0.3-30 nmol elicited dose-related increases in cavernosal pressure, which, at the highest dose studied, were comparable to increases induced by the triple-drug standard (papaverine, phentolamine, and prostaglandin E1), a preparation used in the treatment of erectile dysfunction. Responses to [Tyr1]-nociceptin were rapid in onset and had a time course similar to responses to nociceptin. Metenkephalin increased cavernosal pressure, whereas injections of nociceptin-(2-17), dynorphin A, and beta-endorphin did not alter cavernosal pressure. Erectile responses to nociceptin and to [Tyr1]-nociceptin were not altered after administration of the opioid receptor antagonist naloxone at a time when erectile responses to metenkephalin were attenuated. These data show that [Tyr1]-nociceptin and nociceptin have similar naloxone-insensitive erectile activity in the cat. PMID- 9876027 TI - Protein kinase CK2 activities in human prostatic and seminal-vesicle secretions and seminal plasma. AB - Human prostatic secretion and seminal plasma contain certain protein kinase activities. Protein kinases play important roles in regulating a vast variety of cellular functions. The objective of this study was to determine whether one of these protein kinase activities in human prostatic secretion and seminal plasma is due to CK2, a messenger-independent, serine/threonine protein kinase that has considerable potential as a regulatory enzyme. By employing an anti-CK2 antibody and a CK2-specific peptide substrate, we have established that CK2 is present in these secretions. Approximately 70% of the CK2 activity present in seminal plasma of normozoospermic men (n = 49) is correlated to the number of sperm originally present in the semen. Further, both the prostate gland and the seminal vesicles are sources of CK2 activity in the seminal plasma of vasectomized men (n = 38). Although there was considerable variation between individuals in CK2 activity, the variation in repeat semen samples of the same vasectomized men (n = 6) was within 21%. There was no correlation of CK2 activity in seminal plasma with age for vasectomized (27-48 years, n = 38), oligozoospermic (28-43 years, n = 24), or normozoospermic men (26-48 years, n = 49). These data suggest that the majority of CK2 activity in the seminal plasma of normozoospermic men originates from sperm but that the prostate and seminal vesicles are accessory sex-gland sources of this enzyme. PMID- 9876028 TI - A pharmacokinetic study of injectable testosterone undecanoate in hypogonadal men. AB - Testosterone undecanoate (TU) provides testosterone (T) replacement for hypogonadal men when administered orally but requires multiple doses per day and produces widely variable serum T levels. We investigated the pharmacokinetics of a newly available TU preparation administered by intramuscular injection to hypogonadal men. Eight patients with Klinefelter's syndrome received either 500 mg or 1,000 mg of TU by intramuscular injection; 3 months later, the other dose was given to each man (except to one, who did not receive the 1,000-mg dose). Serum levels of reproductive hormones were measured at regular intervals before and after the injections. Mean serum T levels increased significantly at the end of the first week, from less than 10 nmol/L to 47.8+/-10.1 and 54.2+/-4.8 nmol/ L for the lower and higher doses, respectively. Thereafter, serum T levels decreased progressively and reached the lower-normal limit for adult men by day 50 to 60. Pharmacokinetic analysis showed a terminal elimination half-life of 18.3+/-2.3 and 23.7+/-2.7 days and showed a mean residence time of 21.7+/-1.1 and 23.0+/-0.8 days for the lower and higher doses, respectively. The area under the serum T concentration-time curve and the T-distribution value related to serum T concentration were significantly higher following the 1,000-mg dose than following the 500-mg dose. The 500-mg dose, when given as the second injection, yielded optimal pharmacokinetics (defined as mean peak T values not exceeding the normal range and persistence of normal levels for at least 7 weeks), suggesting that repeated injections of 500 mg at 6-8-week intervals may provide optimal T replacement. The mean serum levels of estradiol were normalized following the injections, and prolactin levels were normal throughout the study. Significant decrease of serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels was observed, with the decrease in LH levels being more pronounced. There were no significant differences in serum LH and FSH levels between the two doses. Sex hormone-binding globulin (SHBG) levels before any T therapy were near the upper limit of normal for adult men and were reduced by approximately 50% just prior to the second dose of TU. The decreased SHBG levels produced by the first TU injection could have led to lower peak total T levels and to a more rapid clearance of T following the second TU injection. We conclude that single-dose injections of TU to hypogonadal men can maintain serum T concentration within the normal range for at least 7 weeks without immediately apparent side effects. It is likely that this form of T would require injections only at 6-8-week or longer intervals, not at the 2-week intervals necessary with currently used T esters (enanthate and cypionate). This injectable TU preparation may provide improved substitution therapy for male hypogonadism and, in addition, may be developed as an androgen component of male contraceptives. PMID- 9876029 TI - The epidemiology of vascular trauma. AB - The study of blood vessel injury has largely focused on diagnosis, treatment, and the outcome of treatment. Although the epidemiology of trauma in general has been extensively studied and reported, the specific epidemiology of vascular injury has not. This report includes a general discussion of the incidence of and risk factors associated with injury to blood vessels. Vascular injury epidemiology has been studied in three different settings: military conflicts, large urban centers, and to a lesser extent, in rural areas. There has been an increase in the incidence of vascular injury in the United States over the past four decades that has paralleled the increase in assault with firearms, motor vehicle crashes, and invasive medical procedures. Mortality and the utilization of medical resources is higher among patients with vascular injury than among trauma patients without blood vessel injuries. There are well-documented differences in the demographics of blood vessel injury in urban versus rural settings. The mechanisms of injury (penetrating, blunt, iatrogenic) are associated with characteristic anatomic patterns of vascular and other organ system injury. PMID- 9876030 TI - Cervicothoracic vascular injuries. AB - Arterial and venous trauma of the cervicothoracic region continues to present challenging problems for the surgeon, despite advances in vascular diagnostics and surgical technique. Whether due to penetrating or blunt mechanisms, overall incidence of these injuries is low, whereas morbidity and mortality remain high. Despite collective experience from busy trauma centers, there still remain controversies regarding diagnostic evaluation, operative approach, and surgical treatment of these potentially devastating injuries. Therefore, this article compares and contrasts recent literature and controversies surrounding the treatment of cervicothoracic trauma. Pros and cons of duplex ultrasonography and angiography in the diagnosis of carotid and vertebral artery injury are highlighted, and selective versus mandatory neck exploration for zone II penetrating injuries are discussed. Increasing awareness of blunt carotid artery injury is emphasized, including management dilemmas that frequently accompany this type of injury. In addition, we review interventional radiological techniques for the management of vertebral artery injury and surgical approaches for aortic arch branch vessel or major cervicothoracic vein injury. PMID- 9876031 TI - Management of abdominal vascular injuries. AB - Major abdominal vascular injuries are associated with significant morbidity and mortality. They present a formidable challenge to the most experienced surgeon. Most commonly the result of penetrating trauma, hemorrhagic shock is the usual presentation, and diagnostic maneuvers are unnecessary. Conversely, a high index of suspicion is critical in diagnosing occult injuries. This article reviews the cause, diagnosis, and treatment of major abdominal vascular injuries, with an emphasis on exposures and techniques of repair. In addition, the concepts of repair versus ligation, staged laparotomy, and the abdominal compartment syndrome are discussed. PMID- 9876032 TI - Changing perspectives in the diagnosis of peripheral vascular trauma. AB - The diagnosis of peripheral vascular trauma has undergone significant evolution in the last two decades. A minority of patients with arterial trauma present with classic findings that make the diagnosis obvious. However, the diagnosis of occult arterial injuries is more challenging, and it is for this group that the diagnostic algorithm has changed most significantly. Because of the low yield of routine operative exploration and routine arteriography to evaluate potential injury to vessels in proximity to penetrating wounds, many authors now recommend the selective use of arteriography or other diagnostic modalities based on the results of clinical examination and noninvasive pressure determinations. This article reviews the evidence in support of such a selective approach to the diagnosis of arterial injuries. PMID- 9876033 TI - Vascular trauma associated with fractures and dislocations. AB - Vascular trauma occurs relatively infrequently in association with general orthopedic trauma but may be seen more often in injuries involving joint dislocations and areas in which vascular structures are tethered at the fracture site. The mechanisms of vascular trauma are identified, and the general principles in managing these injuries, including the operative approach to the injury itself and the options in repairing both arterial and venous injuries, are discussed. The role of fasciotomy and primary amputation are also reviewed. Several specific injuries, including pelvic fracture, knee dislocation, shoulder and upper limb injuries, complex tibial fractures, and iatrogenic injuries, are examined from diagnostic and management perspectives. Despite an ongoing evolution in the diagnosis and management of these often challenging injuries, the essential requirements for a good clinical outcome remain early recognition of the dysvascular limb and rapid institution of therapy. PMID- 9876034 TI - Deep venous thrombosis in the trauma patient. AB - Despite improvements in trauma care and thromboembolism prophylaxis over the past 50 years, deep venous thrombosis (DVT) remains a source of morbidity and mortality in the injured patient. Pulmonary embolism may complicate the course of up to 6% of such patients, although the morbidity associated with late development of the postthrombotic syndrome remains poorly defined and perhaps underemphasized. Prolonged immobilization and venous injury contribute to the risk of venous thrombosis, and accumulating evidence suggests underlying imbalances between activated coagulation and fibrinolysis. Much investigative work has focused on identifying subpopulations, including older patients (30 to 40 years) and those with prolonged immobilization, pelvic and lower extremity fractures, spinal cord injury, major venous injury, central venous lines, significant blood requirements, and admission coagulopathy, who are at particular risk of developing DVT. Given the diverse underlying pathophysiologic factors, it is perhaps not surprising that the benefits of prophylactic measures have been imperfect and remain controversial. Both intermittent pneumatic compression and low-dose unfractionated heparin have limitations, although the low-molecular weight heparins have promise for prophylaxis in this population. It has also been suggested that other measures, such as serial duplex ultrasound screening and prophylactic vena cava filters, may have a role in very high-risk patients and those who are not candidates for other means of prophylaxis. However, the precise role of these measures remains poorly defined and awaits prospective validation. An improved understanding of the operative risk factors may permit prophylactic therapy to be better tailored to the individual patient. PMID- 9876035 TI - Iatrogenic vascular trauma. AB - With the increasing performance of percutaneous transluminal angioplasty and insertion of an increasing number of intravascular devices, the size of arterial punctures has been increasing. A consistent minority of these procedures will result in vascular injuries requiring treatment. At the same time, the regionalized nature of trauma care in the United States has resulted in a large number of vascular surgeons who are exposed to vascular trauma only when iatrogenic. The most common injuries observed are caused by percutaneous vascular instrumentation and include hemorrhage and pseudoaneurysm that may compress adjacent structures, fistula, acute occlusion, and embolization. Injuries unique to balloon angioplasty/stenting include arterial rupture and dissection. Indwelling intravascular devices are another common source of iatrogenic vascular injury ranging from arterial rupture to thrombosis and embolization. Much less common injuries are observed in orthopedic and abdominal/laparoscopic operations but show reproducible causes/patterns. Finally, pediatric iatrogenic vascular trauma is relatively common because of the small size of the vasculature, but the natural history and management is markedly different from that in adults. PMID- 9876036 TI - Compartment syndrome: new insights. AB - Long recognized as a potential problem in trauma patients or victims of acute extremity ischemia, compartment syndrome is seen in other patient populations as well. In such patients, the absence of timely diagnosis and effective treatment inevitably results in loss of extremity neuromuscular function. New insights into the pathophysiology of compartment syndrome provide opportunities for earlier and more accurate recognition as well as the possibility that the morbid consequences of the syndrome might be mitigated or in certain settings even prevented. PMID- 9876037 TI - Neuronal nitric oxide synthase is permanently decreased in the cerebellum of rats subjected to chronic neonatal blockade of N-methyl-D-aspartate receptors. AB - Pharmacological blockade of the (NMDA) receptor at critical stages of brain development may have long-lasting effects on brain chemistry and on animal behavior. We report here experiments in which the competitive NMDA receptor antagonist CGP 39551 was administered to rat pups from postnatal day 7 (P7) to P18. The stage of treatment was selected to primarily target the cerebellum, whose granule cells undergo post-mitotic migration and establishment of synaptic connections during this period. We focused our study on the long-term consequences of CGP 39551 treatment on the neuronal isoform of nitric oxide synthase (nNOS) since nNOS is highly expressed in the cerebellum and it is functionally linked to the NMDA receptor. Treated rats exhibited a long-lasting (up to P70) decrease in the intensity of nNOS immunocytochemical staining in the cerebellar cortex accompanied by a decrement of calcium-dependent NOS catalytic activity. A comparable decrease of enzyme activity was measured in the cerebral cortex, but not in the hippocampus, of adult rats. Other neurochemical markers (glutamatergic, gabaergic, purinergic) and glutamine synthetase were unchanged, while a cholinergic marker was slightly increased in the cerebellum of CGP 39551 treated animals. Taken together these data show that blockade of NMDA receptor during the critical period of formation and stabilization of neuronal circuits preferentially affects long-term nNOS expression and catalytic activity in the cerebellum. PMID- 9876038 TI - Effect of oral administration of Pinellia ternata, Zingiberis rhizoma and their mixture on the efferent activity of the gastric branch of the vagus nerve in the rat. AB - The effect of taste stimulation of Pinellia temata, Zingiberis rhizoma and their mixture on the efferent activity of the gastric branch of the vagus nerve was observed in the anesthetized rat. Taste stimulation by Pinellia ternata (50 mg/ml, 10 min) resulted in a suppression in vagal gastric nerve activity. On the contrary, stimulation by Zingiberis rhizoma (50 mg/ml, 10 min) caused a facilitation in efferent activity. The mixture of Pinellia ternata and Zingiberis rhizoma (5:1, 50 mg/ml, 10 min stimulation) demonstrated no suppressive effect on gastric nerve activity. These observations indicate that it is reasonable to prescribe Pinellia ternata with Zingiberis rhizoma in traditional Japanese medicine to prevent suppressive effect of the taste of Pinellia ternata on gastric function. PMID- 9876039 TI - Hemispheric lateralization in preattentive processing of speech sounds. AB - Event-related magnetoencephalographic (MEG) responses to infrequently presented spoken deviant syllables [di] and [ba] among repetitive standard [da)]syllables were recorded in subjects who either attended to these stimuli in order to discriminate the [ba] syllables or ignored them while attending a silent movie. In both conditions, the deviant syllables elicited a mismatch response (MMNm, the magnetic counterpart of mismatch negativity), which was stronger in the left than in the right auditory cortex, indicating left-hemispheric dominance in speech processing already at a preattentive processing level. PMID- 9876040 TI - Reduction of cholecystokinin-8S-neurons in the paraventricular hypothalamic nucleus of neonatally overfed weanling rats. AB - Cholecystokinin (CCK) is suggested to be involved, e.g. in the central nervous modulation of food intake, possibly by acting within specific hypothalamic nuclei. Perinatal overnutrition predisposes to permanent obesity and hyperphagia, while underlying mechanisms are unclear. By reducing the litter size from the 3rd to 21st day of life, early overnutrition was induced in newborn rats. At weaning, clear overweight (P < 0.001), hyperglycaemia (P < 0.05), hyperinsulinaemia (P < 0.001), and insulin resistance (P < 0.001) occured. These early signs of obesity were associated with a significantly decreased number of CCK-positive neurons in the paraventricular hypothalamic nucleus (P < 0.002). In conclusion, due to neonatal overfeeding malformation of CCKergic neurons at the end of the critical hypothalamic differentiation period occurs. Long-term consequences on CCK-related neuroendocrine regulations could be suggested, including those affecting food intake and body weight gain. PMID- 9876041 TI - Dizocilpine and reduced body temperature do not prevent methamphetamine-induced neurotoxicity in the vervet monkey: [11C]WIN 35,428 - positron emission tomography studies. AB - [11C]WIN 35,428 (WIN), a cocaine analog that binds to the dopamine transporter (DAT), and positron emission tomography (PET) were used to evaluate the potential neuroprotective effects of dizocilpine (MK-801) on methamphetamine (MeAmp) induced neurotoxicity in the striatal dopamine system of the vervet monkey. MK 801 (1 mg/kg, i.m.) was administered 30 min prior to a neurotoxic MeAmp dosage for this species (2 x 2 mg/kg, 4 h apart); control subjects received MeAmp. MK 801 treated subjects were anesthetized by the drug for 6-8 h; throughout that period, a 2-3 degrees C decrease in body temperature was measured. At 1-2 weeks post-MeAmp, decreases of approximately 75% in striatal WIN binding were observed for both MK-801/MeAmp and MeAmp subjects. Thus, in this non-human primate species, the combination of MK-801 pretreatment and reduced body temperature did not provide protection from the MeAmp-induced loss of DAT. Further, the absence of an elevated body temperature during the acute MeAmp exposure period indicated that hyperthermia, per se, was not a necessary concomitant of the MeAmp neurotoxicity profile as has been previously demonstrated in rodents. These results provide evidence that different regulatory factors maintain the integrity of the rodent and primate striatal dopamine systems. PMID- 9876042 TI - Differentiation-specific mRNA expression of a mouse bipotential glial cell line. AB - We have previously reported that a bipotential glial cell line from mouse cerebrum, designated OS3, phenotypically differentiates into oligodendrocytes and astrocytes both in vitro and in vivo. To study the potential mechanisms of differentiation, in this study we investigated mRNA expression of cytokines and developmentally regulated proteins in OS3 during differentiation into oligodendrocytes by semi-quantitative reverse transcription and polymerase chain reaction. In the presence of 10% calf serum OS3 cells expressed IL-1alpha and IL 1beta mRNA. However, when the cells were cultured in chemically defined medium or low serum-containing medium the expression of IL-1alpha and IL-1beta mRNA was down-regulated. Under stimulation of phorbol ester, expression of IL-6 and nerve growth factor mRNA was up-regulated. The capacity for differentiation of OS3 cells into oligodendrocytes in vitro was limited and most OS3 cells ceased their differentiation at the proligodendroblast stage. However, expression of proteolipid protein (PLP) and DM20 mRNA was detectable and was up-regulated in accordance with the differentiation into oligodendrocytes. As a control, primary astrocytes expressed DM20 mRNA but not PLP mRNA and the expression of DM20 mRNA was independent of culture condition. Therefore, OS3 cells will be of use for the study of differentiation of progenitor cells into type-2 astrocytes or oligodendrocytes at the molecular level. PMID- 9876043 TI - Inhibition of brain choline uptake by isoarecolone and lobeline derivatives: implications for potential vector-mediated brain drug delivery. AB - Delivery of certain compounds to brain is restricted by the nature of the blood brain barrier (BBB). Many valuable pharmaceuticals are excluded from the CNS due to hydrophilicity or charge. These limitations have been overcome by numerous methods. One method we use is to take advantage of saturable nutrient transporters located at the barrier. These systems transport hydrophilic and charged nutrients into brain such as choline, a quaternized neurotransmitter precursor. Using knowledge of their substrate specificity, it is possible to deliver agents into brain using these nutrient carriers. In this report, derivatives of lobeline and isoarecolone were evaluated to determine if they may gain access to brain by the blood-brain barrier basic amine transporter using the in situ brain perfusion technique. These compounds do bind the blood-brain barrier basic amine transporter and may enter brain by this transport system. PMID- 9876044 TI - Effect of systemic ethanol on basal and stimulated glutamate releases in the nucleus accumbens of freely moving Sprague-Dawley rats: a microdialysis study. AB - This study was conducted to assess the impact of systemic ethanol (EOH) on the glutamatergic transmission in the nucleus accumbens (NACC). Extracellular concentrations of glutamate (GLU) in the NACC of freely moving Sprague-Dawley rats were monitored by intracerebral microdialysis. Intraperitoneal injection of EOH at a dose of 2 g/kg significantly decreased basal extracellular GLU by 21%. In addition, administration of the same dose of EOH significantly attenuated 150 mM K+-stimulated GLU release from the NACC by more than 50%. Since K+-stimulated GLU release has been demonstrated to derive largely from nerve terminal depolarization, reductions of K+-evoked GLU release may reflect in part the effect of EOH on the neurotransmitter pool. The present results suggest that EOH may suppress glutamatergic transmission in the NACC by lowering presynaptic GLU release. PMID- 9876045 TI - Dopamine and acetylcholine elevation on lever-press acquisition in rat prefrontal cortex. AB - To determine whether the rat medial prefrontal cortex (PFC) is involved in acquiring operant learning, we observed changes in extracellular concentration of dopamine (DA) and acetylcholine (ACh) in the rat medial PFC during lever-press acquisition (acquisition group) or retrieval (retention group) using in vivo microdialysis. We found that DA or ACh elevation related to acquisition occurred. DA elevation was observed in the acquisition group only. These results indicate that the medical PFC is related to acquisition, and suggest that interaction between DA and ACh may be involved in learning acquisition. PMID- 9876046 TI - Rat embryonic hippocampal neurons express a new class A calcium channel variant. AB - The aim of this study was to investigate, using a RT-PCR strategy, rat voltage gated class A calcium channel (alpha1A) splice variants during rat hippocampus development. Results demonstrate the presence of multiple alpha1A mRNAs with the hippocampus formation and revealed a new variant of the rat alpha1A subunit (alpha1A-EFe) that diverges from alpha1A-a in the EF-hand domain. alpha1A-EFe expression in hippocampal neurons is restricted to the embryonic period. This in vivo developmental program is recapitulated in dissociated cultures of E17 embryonic hippocampal neurons. These data demonstrate that rat hippocampus neurons express a unique alpha1A splice variant during the embryonic period and suggest that alternative RNA splicing may modulate neuronal calcium channel properties during development. PMID- 9876047 TI - Expression of calcitonin receptor-like receptor and receptor-activity-modifying proteins in human cranial arteries. AB - Recently a new type of proteins modulating the pharmacological profile of the calcitonin receptor-like receptor (CRLR) were identified. The receptor-activity modifying proteins (RAMPs) were shown to be essential for the expression of a functional CRLR and furthermore the RAMPs seemed to modify ligand selectivity of CRLR: coexpression of CRLR and RAMP1 resulted in a CGRP1 type of receptor while an adrenomedullin receptor resulted when CRLR and RAMP2 were coexpressed. In the present study significant molecular expression of CRLR concomitant with RAMP1, 2 and 3 were demonstrated in human meningeal, cerebral and temporal arteries by use of reverse transcriptase polymerase chain reactions (RT-PCR). These findings support previous studies demonstrating functional CGRP1 receptors in human cranial arteries. Furthermore the present study suggests the potential for functional adrenomedullin receptors in human cranial arteries. PMID- 9876048 TI - Gap junction uncoupling of neonatal rat astroglial cultures does not affect their expression of fibroblast growth factor 2. AB - Fibroblast growth factor (FGF)-2 is expressed by astroglial cells and regulates astroglial proliferation, differentiation, and gap junction communication. We show now that uncoupling of astroglial cells cultured from neonatal rat cortex, striatum and mesencephalon by 18alpha-glycirrhetinic acid (AGA) or arachidonic acid (AA) (demonstrated by intracellular injections of Lucifer Yellow) does affect neither levels of FGF-2 mRNA (investigated by a competitive RT-PCR method), nor protein (as revealed by Western blot analysis). This suggests that expression of FGF-2, which reduces gap junction communication of cortical and striatal astroglial cells [15], is not affected when astroglial cells are uncoupled. Regulation of FGF-2 expression and coupling in astroglial cells are thus not reciprocal suggesting more complex interrelationships between these two important parameters of astroglial cell differentiation. PMID- 9876049 TI - Inspiratory inhibition of pulmonary rapidly adapting receptor relay neurons in the rat. AB - Second-order relay neurons (referred to as rapidly adapting pulmonary stretch receptors (RARs)-cells) activated by vagal afferents originating from RARs were identified electrophysiologically in Nembutal-anesthetized, paralyzed and artificially ventilated rats. Their location in the commissural subnucleus of the nucleus tractus solitarii (NTS) and firing properties in response to lung inflations and deflations were studied in detail for the first time in the rat. Importantly, in all the RAR-cells examined, their firing was suppressed to various extent during a period from inspiration to early expiration. This means that the information from the RARs to the central nervous system is gated by the central respiratory system at the level of RAR-cells in the NTS. PMID- 9876050 TI - Non-N-methyl-D-aspartate receptors may mediate the transmission of emetic signals between visceral vagal afferents and the solitary nucleus in dogs. AB - The effects of the N-methyl-D-aspartate (NMDA) and non-NMDA receptor antagonists MK-801 and NBQX, respectively, on salivary secretion and retching induced by vagal stimulation were studied in decerebrate dogs. Vagal stimulation induced an increase in salivary secretion and fictive retching. Intra-4th ventricular application of vehicle or MK-801 did not change either response, while NBQX completely abolished both responses. These results suggest that non-NMDA receptors mediate both responses in the solitary nucleus. PMID- 9876051 TI - The effect of calcium channels blockers in the K+-evoked release of [3H]adenine nucleotides from rat brain cortical synaptosomes. AB - The role of L-,N-, P- and Q-type voltage-dependent calcium channels in K+-induced release of [3H]adenine nucleotides from rat brain cortical synaptosomes was investigated. Cd2+, a non-specific blocker of calcium channels, inhibited by 69% the release of the nucleotides induced by 33 mM K+. Nifedipine, omega-Conotoxin GVIA and omega-Agatoxin IVA had no effect whereas omega-Conotoxin MVIIC inhibited by 62% the K+ induced release of adenine nucleotides in rat brain cortical synaptosomes. It is concluded that Q-type calcium channels are directly involved in the release of adenine nucleotides in rat brain cortical synaptosomes. PMID- 9876053 TI - Extended-spectrum beta-lactamases in clinical isolates of Klebsiella pneumoniae from Zagreb, Croatia. AB - Forty clinical isolates of Klebsiella pneumoniae, from various clinical specimens, with reduced susceptibility to ceftazidime, were tested for extended spectrum beta-lactamase (ESBL) production. ESBL production was demonstrated by an 8-fold reduction in the minimum inhibitory concentration (MIC) of ceftazidime combined with clavulanate (2 mg/L) compared to ceftazidime alone in all strains. The aim of this investigation was the biochemical and molecular characterization of the ESBL produced by K. pneumoniae strains and their Escherichia coli transconjugants. Transfer of ceftazidime resistance was demonstrated in 23 of 40 strains. Thirteen strains produced an ESBL with the isoelectric point of 8.2 which was encoded by a self-transferable multiresistance plasmid of 150 kb. The substrate profile was similar to that of the SHV-5 isolated initially in Chile. Seven of these 12 strains had an additional TEM beta-lactamase. Six isolates and their transconjugants produced a plasmid-encoded ESBL with an isoelectric point close to 5.4. The remaining 21 strains produced an ESBL with an isoelectric point of 7.6 (thus probably SHV-2) which was encoded on a plasmid transferable to E. coli in 4 strains only. Four of those strains possessed an additional plasmid encoded TEM beta-lactamase with an isoelectric point close to 5.4. The transconjugants harbored a multiresistance plasmid of 150 kb. Thus SHV-2 and SHV 5 enzymes appear to have been the most common ESBLs in K. pneumoniae from Zagreb during 1994-1995. PMID- 9876052 TI - Antibiotic-induced release of bacterial cell wall components in the pathogenesis of sepsis and septic shock: a review. AB - This article reviews the new criteria for selecting the proper antimicrobial agent and dosage regimen for standard treatment of severe sepsis, with the intention of preventing septic shock. After introducing new concepts on the pathogenesis of sepsis and septic shock, the authors analyze the parameters of betalactam antibacterial activity, the antibiotic-induced release of bacterial endotoxin and the interrelationships between pharmacokinetics and pharmacodynamics of antibiotics in the search for an optimum dosage regimen of antimicrobial mono- or polytherapy for severely ill septic patients admitted to the intensive care unit. PMID- 9876054 TI - A randomized, observer-blind trial of amoxycillin/clavulanate versus cefaclor in the treatment of children with acute otitis media. Augmentin 415 Study Group. AB - In this randomized, multicenter, observer-blind study, the efficacy, safety and tolerability of amoxycillin/clavulanate and cefaclor were compared in children with a clinical diagnosis of acute otitis media. Patients aged between 1 and 12 years received either amoxycillin/clavulanate (250 mg/62 mg t.i.d., or 125 mg/31 mg t.i.d. if aged under 6 years) or cefaclor (250 mg t.i.d., or 125 mg t.i.d. if aged under 6 years) for 7 days. The amoxycillin/clavulanate regimen was based on a dose of 20/5 mg/kg/day (representing 20 mg amoxycillin plus 5 mg clavulanic acid) in three divided doses. Patients were followed-up at the end of therapy and on days 10-12 and 38-40. At the end of the study (days 38-40), clinical success rates were 91.4% for amoxycillin/clavulanate and 78.6% for cefaclor. The difference was statistically significant (p = 0.008). After the 7 days of treatment, 3 patients (2.9%) in the amoxycillin/clavulanate group had clinical failure, compared with 18 patients (16.1%) in the cefaclor group (p < 0.001). Both treatments were well tolerated and there were no statistically significant differences between the groups in adverse event profiles. The incidence of diarrhea was low (7.0% amoxycillin/clavulanate, 8.4% cefaclor) and was generally of mild or moderate intensity. The study demonstrated that amoxycillin/clavulanate was significantly more effective clinically than cefaclor in the treatment of acute otitis media in children. PMID- 9876055 TI - Azithromycin compared with minocycline in the treatment of acne comedonica and papulo-pustulosa. AB - This open study was conducted in 72 outpatients with acne vulgaris, to compare the clinical efficacy and tolerability of azithromycin and minocycline. Azithromycin was administered as a single oral dose (500 mg/day) for 4 days in four cycles every 10 days and minocycline was administered 100 mg daily for 6 weeks. Improvement was assessed 6 weeks after initiation of treatment with a four graded scale. A satisfactory clinical response was observed in 75.8% of the patients treated with azithromycin and in 70.5% of those treated with minocycline. There were no significant differences between these two acne treatments in terms of reduction of the number of lesions (p> 0.05). Both agents were well tolerated and mild side effects were reported in 10.3% of azithromycin and 11.7% of minocycline treated patients. We conclude that azithromycin is at least as clinically effective and well tolerated as minocycline as treatment of facial comedonic and papulopustular acne. PMID- 9876056 TI - Long term treatment with clarithromycin for cryptosporidiosis and emergence of drug resistant disseminated infection due to Mycobacterium avium: case report. AB - Macrolide resistance in disseminated Mycobacterium avium infection is of major concern in AIDS patients as these drugs represent the main component of combination therapy. Clarithromycin and azithromycin should not be used alone for the treatment and prophylaxis of the disease because of the risk of selecting resistant strains. We report the case of a clarithromycin resistant disseminated M. avium infection in an AIDS patient, acquired after long term monotherapy with clarithromycin for the treatment of cryptosporidiosis. PMID- 9876057 TI - Multimodal treatment strategy for locally advanced breast cancer. AB - Between 1990 and 1995, 87 patients with locally advanced breast cancer were treated with initial chemotherapy consisting of 3-4 cycles of epirubicin and then with surgery, radiotherapy and chemotherapy. All patients with positive or unknown estrogen receptor status were administered tamoxifen. A complete response was observed in 9 patients and a partial response in 64. At a mean follow-up of 29.22 months, 25 patients had died of metastatic disease, and 48 patients are disease-free. 54% patients are alive at 5 years. Statistical analysis confirmed that neither age, menopausal status, size of the primary tumor nor histology seemed to influence the Disease Free and Overall Survival. Improved survival was observed in patients with negative lymph nodes and positive receptor status. The presence of a positive receptor status may be correlated with conserved integrity of hormone sensitivity and with a good response to the hormone therapy. PMID- 9876058 TI - Ifosfamide and actinomycin-D, added in the induction phase to vincristine, cyclophosphamide and doxorubicin, improve histologic response and prognosis in patients with non metastatic Ewing's sarcoma of the extremity. AB - The role of ifosfamide as first-line chemotherapy treatment of non metastatic Ewing's sarcoma of the extremity is still under discussion. The purpose of this paper is to report the results achieved in a neoadjuvant protocol (REN-3) in which ifosfamide, added to the conventional VACA regimen, was employed since the induction phase. Induction chemotherapy consisted of vincristine, cyclophosphamide, doxorubicin, actinomycin-D and ifosfamide. After local treatment, patients received the drugs used in the induction phase and etoposide. Between November 1991 and November 1994, 61 patients with non metastatic Ewing's sarcoma of the extremity were treated. Forty-nine patients underwent surgery and 73.5% of them had a good histologic response. At a median follow-up of 60 months (range 32-76), 48 patients (79%) remained continuously disease-free. The 5-year event-free and overall survival were 77% and 87%, respectively. These results were significantly better both in terms of histologic response or event-free and overall survival than those obtained in 58 patients with non metastatic Ewing's sarcoma of the extremity treated in a previous protocol (REN-2) in which the same drugs were used, but ifosfamide was employed only in the maintenance phase. The present study suggests the importance of early use of ifosfamide in the treatment of patients with non metastatic Ewing's sarcoma of the extremity. PMID- 9876059 TI - Phase II study with ifosfamide, carboplatin, etoposide (ICE regimen) at intermediate dosage for advanced non small cell lung cancer (NSCLC). AB - We have evaluated the combination of ifosfamide, carboplatin and etoposide (ICE regimen) along with mesna in 26 previously untreated patients with non small cell lung cancer (NSCLC). Thirteen stage III B and 13 stage IV patients received intermediate doses of ifosfamide (1000 mg/m2), carboplatin (120 mg/m2) and etoposide (120 mg/m2) given intravenously on day 1 to 3 every 4 weeks. Except for one patient who experienced grade 3 transient thrombocytopenia no major events of hematological or systemic toxicity were observed. Response rate (27%, 95% C.I., 10 to 44%), median duration of response (9 months, range 6-15), and survival (9.5 months, range 2-44+) were comparable to those achieved with conventional cisplatin-containing regimens. Our ICE combination, as compared to standard or high dose schedules appears effective, safe, well tolerated, and devoid of severe hematological toxicity. PMID- 9876060 TI - Vascular gene therapy. PMID- 9876061 TI - Radionuclide scanning in parathyroid diseases. AB - BACKGROUND: New radionuclide agents and scanning procedures have markedly improved preoperative parathyroid gland localization. The aim of this review was to evaluate the results of parathyroid scanning and to clarify its current place in the clinical management of parathyroid diseases. METHODS: The literature describing the different radioisotopes and available scanning techniques was analysed and their clinical outcome in various parathyroid diseases was evaluated. RESULTS: Using 99mTc-radiolabelled sestamibi or other 9mTc-labelled cationic complexes, parathyroid scintigraphy detects 87 per cent of solitary adenomas (n=894), 55 per cent of abnormal glands in patients with multiglandular disease (n=303) and 75 per cent of persistent or recurrent lesions in patients with previous neck surgery (n=240). If necessary, three-dimensional imaging techniques can clarify the spatial localization of an ectopic uptake focus. The positive predictive value of the available scanning procedures is over 95 per cent, but false-positive uptake may be encountered. CONCLUSION: Radionuclide parathyroid gland scanning with 99mTc-labelled cationic complexes is the initial non-invasive method of choice for preoperative parathyroid gland localization in patients at special risk and/or with previous neck exploration. While scanning has also been proposed to facilitate parathyroidectomy, there is no objective evidence to support its routine use before a first-time surgical procedure. PMID- 9876062 TI - Solitary rectal ulcer syndrome. AB - BACKGROUND: Solitary rectal ulcer syndrome is a rare disorder characterized by erythema or ulceration of the rectal wall, associated with typical histological features, and disturbed defaecatory behaviour with the passage of blood and mucus. METHODS: This is a review based on a literature search using a computer database (Medline) and manual cross-referencing. RESULTS: The pathogenesis is likely to vary in different patients; it includes trauma from straining, direct digital trauma and possibly primary neuromuscular pathology. The histological findings of extension of the muscularis mucosa between crypts and muscularis propria disorganization on full-thickness specimens are characteristic. Biofeedback defaecation retraining, including habit training, can lead to symptom improvement and return to work in a majority of patients. Abdominal rectopexy offers long-term symptom improvement in approximately 50 per cent of patients. Rectal ulceration may persist after any treatment, even if symptoms improve. CONCLUSION: Behavioural therapy and carefully considered operations offer the best treatment results. Further work on psychological factors and neuromuscular and vascular pathology is required. PMID- 9876063 TI - Influence of study design on reported mortality and morbidity rates after abdominal aortic aneurysm repair. AB - BACKGROUND: The mortality and morbidity rates of elective abdominal aortic aneurysm (AAA) surgery, as reported over the past 12 years, were graded and analysed by levels of evidence. METHODS: Articles on elective AAA surgery published between 1985 and 1996 were retrieved and classified into five levels of evidence. Level 1 contains prospective studies and is subdivided into population based (level 1a) and hospital-based (level 1b) studies. Level 2 includes retrospective studies, subdivided into population-based studies (level 2a), hospital-based studies (level 2b) and hospital-based studies concerning a specified group of selected patients (level 2c). Operative mortality and systemic and local/vascular complication rates with 95 per cent confidence intervals were calculated for each level of evidence. RESULTS: Seventy-two articles describing a total of 37 654 patients could be included: two level 1a studies (692 patients), nine level 1b studies (1677 patients), 13 level 2a studies (21 409 patients), 32 level 2b studies (12019 patients) and 16 level 2c studies (1857 patients). The mean 30-day mortality rates of the two population-based levels were similar: 8.2 (95 per cent confidence interval 6.4-10.6) per cent for the prospective (la) and 7.4 (7.0-7.7) per cent for the retrospective (2a) series. These figures were significantly higher than the remarkably similar hospital-based mortality rates: 3.8 (3.0-4.8) per cent for the prospective (1b), 3.8 (3.5-4.2) per cent for the retrospective (2b) and 3.5 (2.8-4.4) per cent for selected patient group (2c) studies. The most frequent complication was of cardiac origin. In the population based series the cardiac complication rates were 10.6 (8.5-13.2) and 11.1 (9.1 13.6) per cent for levels 1a and 2a respectively. This compared well with 12.0 (10.5-13.9) per cent for the prospective hospital-based series (level 1b). The cardiac complication rates in the retrospective hospital-based studies were significantly lower: 8.9 (8.4-9.5) and 6.1 (4.9-7.6) per cent for levels 2b and 2c respectively. CONCLUSION: There is a clear and consistent disagreement in reported mortality rates between hospital-based and population-based studies of elective surgery for AAA. Prospective studies give the best documentation of postoperative morbidity. PMID- 9876064 TI - Plasma concentration of cytokine antagonists in patients with infection following liver resection. AB - BACKGROUND: The interactions between endogenous cytokine antagonists, surgical injury and perioperative infection are poorly understood. METHODS: Perioperative plasma concentrations of soluble tumour necrosis factor receptor p55 (STNFR p55), interleukin (IL) 1 receptor antagonist (IL-1ra) and IL-6 were measured by immunoassay in 29 consecutive patients undergoing liver resection. RESULTS: Plasma concentrations of STNFR p55, IL-1ra and IL-6 showed a striking increase immediately after operation (day 0) and on postoperative day 1. STNFR p55 concentrations correlated significantly with operating time (P< 0.05); IL-1ra levels with operating time, blood loss and weight of resected liver (P < 0.05); and IL-6 concentrations with operating time and blood loss (P< 0.05). Eight of the 29 subjects developed postoperative infections, of whom four had positive cultures of enteric organisms. Plasma STNFR p55 and IL-6 concentrations were significantly higher in the infected group than in the uninfected group (P < 0.05). STNFR p55 and IL-6 concentrations on day 0 differentiated between the two groups with a diagnostic accuracy of 79 and 83 per cent respectively. CONCLUSION: These results suggest that STNFR p55, IL-1ra and IL-6 are produced during operation in response to surgical stress. This production of STNFR p55 and IL-6 may be enhanced further by intraoperative bacterial translocation. Measurement of plasma STNFR and IL-6 concentrations may predict the development of postoperative infectious complications. PMID- 9876065 TI - Amelioration of liver injury by ischaemic preconditioning. AB - BACKGROUND: Ischaemic preconditioning, i.e. preparatory brief ischaemia before subsequent long ischaemia, can effectively protect the heart from ischaemia reperfusion injury in animals. The purpose of this study was to demonstrate the same phenomenon in the liver. METHODS: Using warm ischaemia-reperfusion of 70 per cent of the liver followed by resection of the non-ischaemic portion in rats, livers with 10 min of ischaemic preconditioning, i.e. 10 min of warm ischaemia and reperfusion, were compared with those that had not been subjected to such a manoeuvre. RESULTS: At 120 min after reperfusion following 40 min of warm ischaemia, the livers with 10 min of ischaemic preconditioning had a significantly lower mean(s.d.) serum alanine aminotransferase level (492(217) versus 1236(695) units/l; P < 0005) and lactic dehydrogenase level (7905(4002) versus 15066(9201) units/l; P< 0.05), as well as a higher bile output (0.12(0.03) versus 0.09(0.04) ml per g liver; P < 0.05) and liver tissue adenosine 5' triphosphate level (78(13) versus 61(11) per cent; P< 0.05) than the control livers. The necrosis rate, histologically defined as the percentage of necrotic area in given liver sections, was reduced significantly by this manoeuvre (mean(s.d.) 1.3(1.3) versus 5.3(1.7) per cent; P< 0.05). CONCLUSION: Ischaemic preconditioning exerts a protective effect on hepatic warm ischaemia-reperfusion injury. Such a manoeuvre may be useful for hepatic resection in the clinical setting. PMID- 9876066 TI - Survival following pancreatic carcinoma: a follow-up study of all cases recorded in Malmo, Sweden, 1977-1991. AB - BACKGROUND: The aim of this follow-up study was to assess whether there has been any increase in the percentage of patients offered attempted curative surgery for pancreatic carcinoma and whether the overall survival rate has improved, during the time period 1977-1991. METHODS: Details of new cases of pancreatic carcinoma arising in the population of Malmo during the study period were retrieved from the Local Tumour Register in Lund, Sweden. In all, 740 patients were found; 575 of these were diagnosed before death. Kaplan-Meier analysis was used to calculate overall survival rates, and Cox regression analysis was used to assess survival in relation to year of diagnosis after adjustment for sex, age at diagnosis and stage of disease. RESULTS: Cytological or histopathological evidence of the disease was given in 95 per cent of cases. The overall 5-year survival rate was 0.5 per cent (three of 575). Curative surgery was attempted in 24 patients (4.2 per cent); the proportion undergoing curative surgery increased in the last part of the study. Two of the 24 patients in this group survived for 5 years. CONCLUSION: The prognosis in pancreatic carcinoma remains dismal. Attempted curative surgery still is the only hope for cure, but the group of patients that can be offered this possibility is very limited. In this study, an increase was found in the proportion of patients who were offered attempted curative surgery, but there was no statistically significant increase in the 5-year survival rate following surgery. PMID- 9876067 TI - Splenic complications in chronic pancreatitis: prevalence and risk factors in a medical-surgical series of 500 patients. AB - BACKGROUND: The prevalence, course, treatment, outcome and risk factors of splenic complications in chronic pancreatitis are poorly documented. METHODS: Patients with splenic complications in a medical-surgical series of 500 consecutive patients with proven chronic pancreatitis prospectively followed up for a mean of 7.0 years were compared with patients without splenic complications. RESULTS: Eleven men (2.2 per cent) with alcoholic chronic pancreatitis (median duration 2 (range 0-5) years) had a splenic complication: intrasplenic pseudocyst (n=5), subcapsular haematoma (n=2) or splenic rupture (n=4). All patients except one underwent splenectomy, five of whom also underwent distal pancreatectomy. There were no deaths. Patients with splenic complications had pancreatic tail necrosis (six of 11 versus 17.4 per cent; P=0.007), distal pseudocyst (six of 11 versus 11.7 per cent; P=0.0009) or splenic vein occlusion (seven of 11 versus 10.8 per cent; P< 0.0001) more frequently than those without. In the 22 patients with distal pseudocyst and splenic vein occlusion, the prevalence of splenic complications was 18 per cent (odds ratio 15.0 (95 per cent confidence interval 4.0-55.7). CONCLUSION: Splenic complications occur early in the course of chronic pancreatitis, are rare and are favoured by splenic vein occlusion and pseudocyst or necrosis of the pancreatic tail. Surgical treatment is usually required. PMID- 9876068 TI - Long-term outcome of necrotizing pancreatitis treated by necrosectomy. AB - BACKGROUND: Long-term functional outcome after operative treatment of necrotizing pancreatitis (NP) has not been studied extensively. METHODS: Pancreatic function, performance status, recurrence of symptoms and other related problems were analysed in 44 consecutive patients successfully discharged from hospital after operative necrosectomy (1983-1995) and followed up completely for a mean of 5 years. RESULTS: Clinical pancreatic insufficiency developed in half the patients. Diabetes mellitus (11 patients), steatorrhoea (six) or both (five) were associated with a mean estimate of 52, 66 and 67 per cent parenchymal necrosis respectively. Normal pancreatic function was associated with 27 per cent parenchymal necrosis (P < 0.05). Diabetes worsened while steatorrhoea tended to improve over time. Abdominal pain and pancreatitis recurred in six and two patients respectively. Performance status worsened in four patients because of recurrent pancreatitis and severe steatorrhoea. Poor long-term performance was associated with a higher Acute Physiology And Chronic Health Evaluation II score on admission (mean 14 versus 9). CONCLUSION: NP has prominent effects on long term pancreatic exocrine and endocrine function in half the patients, but most preserve a good overall functional status. The development of pancreatic insufficiency varies with the extent of pancreatic parenchymal necrosis. PMID- 9876069 TI - Surgical treatment and outcome in patients with a hepatocellular carcinoma greater than 10 cm in diameter. AB - BACKGROUND: Hepatocellular carcinoma (HCC) over 10 cm in diameter at the time of diagnosis continues to account for a number of patients undergoing hepatic resection. This study evaluated the clinicopathological features and outcome following surgery for large HCC. METHODS: Forty patients with a large HCC (greater than 10 cm) (group 1) resected between 1991 and 1996 were studied retrospectively. They were compared with 245 patients who had smaller HCCs (10 cm or less) (group 2). RESULTS: No patient in group 1 had hepatitis C infection compared with 22.9 per cent in group 2 (P=0.001). Patients in group 1 were significantly younger, had higher alpha-fetoprotein levels (16750 versus 1864 ng/ml; P < 0.001), better liver function, a higher incidence of multiple tumours (27 of 40 versus 42.0 per cent; P=0.003) and venous invasion (35 of 40 versus 52.2 per cent; P < 0.001), and underwent more major resections (37 of 40 versus 26.5 per cent; P < 0.001) than those in group 2. Morbidity and mortality rates and hospital stay were comparable in the two groups. For group 1, the 1-, 3- and 5-year disease-free survival rates were 42, 30 and 28 per cent respectively. Multiple tumours, venous invasion and impaired liver function were factors associated with recurrence. CONCLUSION: Large HCC had specific clinicopathological features. In selected patients, resection is safe and offers the chance of long-term disease-free survival. PMID- 9876070 TI - Circulating interleukin 6 concentrations and insulin resistance in patients with cancer. AB - BACKGROUND: The mechanism of insulin resistance in patients with cancer is not clear. This study was conducted to evaluate the possible role of circulating cytokines in inducing insulin resistance in patients with cancer. METHODS: Twenty three patients with a variety of cancers were studied, including one patient with oesophageal cancer, 12 with gastric cancer, four with colon cancer and six with lung cancer. Six normal volunteers served as controls. Insulin resistance was evaluated by euglycaemic hyperinsulinaemic glucose clamp with a high physiological insulin concentration of 100 microunits/ml. Metabolized glucose, the M value, was compared between patients with cancer and controls. Serum concentrations of interleukin (IL) 6 and other cytokines (tumour necrosis factor (TNF) alpha, IL-8 and IL-10) were measured by enzyme-linked immunosorbent assay. RESULTS: The mean(s.d.) M value for patients with cancer (5.47(1.59) mg per kg per min) was significantly lower than that for controls (8.23(0.79) mg per kg per min) (P< 0.001). There was no relationship between the M value and degree of body weight loss. Serum IL-6 concentration was measurable in eight of 23 patients: four with lung cancer, two with gastric cancer, and one each with oesophageal and colon cancer. None of the controls had a measurable serum IL-6 concentration. There was no significant relationship between serum IL-6 concentration and body weight loss. TNF-alpha was undetectable in the serum of both patients with cancer and controls. Serum IL-8 and IL-10 were detected in seven and one of 23 patients respectively. These cytokines were not detected in the serum of controls. The M value was significantly smaller in those with measurable serum IL-6 (4.01(1.22) mg per kg per min) than in those with no measurable IL-6 (6.26(1.16) mg per kg per min) (P< 0.001). IL-6 and IL-8 levels were raised more frequently in the same patient but there was no significant relationship between IL-8 and M values. CONCLUSION: These results may suggest that IL-6 is related to insulin resistance in patients with cancer. PMID- 9876071 TI - Nitric oxide generation is increased in experimental renal warm ischaemia reperfusion injury. AB - BACKGROUND: Nitric oxide has a clearly defined place in normal renal homoeostasis while there is a continuing debate as to its role under pathophysiological conditions. This study investigated the role of nitric oxide in a model of renal warm ischaemia-reperfusion injury. METHODS: Groups of rats underwent bilateral renal warm ischaemia (for 15-60 min) followed by reperfusion (20 or 80 min) before unilateral nephrectomy for measurement of renal nitric oxide (as nitroxides) and oxidative damage. Renal function was measured on days 2 and 7 before killing and nephrectomy. A further group received the nitric oxide synthase inhibitor N(G)-nitro L-arginine methyl ester (L-NAME; 50 mg per kg body weight) before induction of warm ischaemia. RESULTS: In early reperfusion there was a correlation between the duration of warm ischaemia (15-45 min) and renal nitrate (r2=0.97) which increased from a mean(s.e.m.) baseline value of 95(5.9) to 208(17.3) nmol per mg protein following 45 min of warm ischaemia. Levels were further raised at 80 min and maintained through to day 7 (241(12.5) nmol per mg protein in 45-min group). This rise was attenuated by L-NAME (P< 0.01) as was the early rise in oxidative damage seen otherwise. By day 7, however, oxidative damage was increased (all P< or = 0.01). CONCLUSION: Renal nitric oxide increased early in recoverable warm ischaemia-reperfusion injury and remained raised to day 7. Nitric oxide synthase inhibition ameliorated early but exacerbated late damage suggesting that the early burst of nitric oxide is cytotoxic but that overall nitric oxide may exert a cytoprotective effect. PMID- 9876072 TI - New model of renal warm ischaemia-reperfusion injury for comparative functional, morphological and pathophysiological studies. AB - BACKGROUND: Renal warm ischaemia-reperfusion injury is pertinent to vascular and transplant surgery. While established models provide functional and morphological data the authors wanted to be able to correlate this with the underlying pathophysiology at any chosen time point, thus allowing future interventional effects on reperfusion injury to be evaluated. METHODS: In a rodent model bilateral renal warm ischaemia (15-60 min) and then reperfusion (20 or 80 min) before nephrectomy allowed for analysis of early reperfusion pathophysiology. The remaining kidney provided functional data (glomerular filtration rate (GFR)) at days 2 and 7 before nephrectomy for late analysis and morphology using a new grading system. RESULTS: Acceptable survival rate (ten of 12 animals) was seen with up to 45 min of warm ischaemia. Renal function was impaired at day 2 following 30-60 min of warm ischaemia (P< 0.01) and day 7 in the 45- and 60-min groups (P < 0.05 and P < 0.01 respectively). Strong correlation existed between duration of ischaemia and GFR at day 2 (r2=0.88) and day 7 (r2=0.95). Histological damage in the cortical tubules was evident in the 45- and 60-min groups (P< 0.01). CONCLUSION: This new model allowed comparative functional, morphological and pathophysiological studies while minimizing the number of animals required. Overall 45 min of warm ischaemia gave significant, recoverable injury and is recommended for investigating renal reperfusion injury. PMID- 9876073 TI - Growth rates and risk of rupture of abdominal aortic aneurysms. AB - BACKGROUND: Early identification of abdominal aortic aneurysms (AAAs) may reduce the risk of death from rupture by providing the opportunity for elective repair. Before a screening policy for AAA is implemented, the growth rates of AAAs and the accompanying risk of rupture without intervention should be established. METHODS: The growth rates of AAAs were calculated using longitudinal aneurysmal growth data from screening studies in Chichester and Huntingdon. Estimates of the growth rates of AAAs and the risks of rupture over time were made taking measurement error and individual variability into account. RESULTS: Growth rate estimates were found to vary by initial aortic diameter, with a more rapid growth seen in large aneurysms (50 mm or more). The rate of aneurysm growth did not differ with age or sex. The estimated risk of rupture of an AAA with an initial diameter of 45 mm did not exceed 20-5 per cent over 5 years. An AAA with an initial diameter of 30 mm has a 4.0 per cent or less chance of rupture over 5 years. CONCLUSION: The study provides a more accurate assessment of the risk of aneurysm rupture without surgery and helps to define rescreening intervals for those with an enlarged aortic diameter. PMID- 9876074 TI - No correlation between phaeochromocytoma catecholamine secretion and granule ultrastructure. AB - BACKGROUND: It is uncertain whether biochemical markers of catecholamine secretion in patients with phaeochromocytoma correlate with tumour ultrastructure granule morphology. METHODS: Fifteen patients with an adrenal phaeochromocytoma (n=13) or paraganglioma (n=2) (three men, 12 women; age 17-79 years) were studied. Catecholamine secretion was estimated by measuring urinary levels of free noradrenaline, adrenaline and dopamine. The number and type of secretory granules were evaluated by two independent observers on electron micrographs (area analysed approximately 70 microm2). Large round or elongated medium-density granules were adrenaline-type granules, whereas electron-dense granules lying in a vacuole were of noradrenaline type. RESULTS: No correlation was found between noradrenaline output and the number or percentage of noradrenaline-type granules, although tumours with normal noradrenaline output had only a minority of this type of granule (less than 25 per cent). Adrenaline-type granules were predominant (77 per cent of 163 granules) in a tumour secreting only adrenaline, but the proportion of adrenaline-type granules in six tumours with normal adrenaline output varied significantly (range 7-89 per cent). It was not possible to evaluate the granule type associated with dopamine secretion because one tumour secreting 14900 nmol dopamine and 1570 nmol adrenaline daily had a predominance of noradrenaline-type granules (63 per cent of 132 granules) and two dopamine-secreting tumours (5500 and 4250 nmol per day respectively) had 93 and 13 per cent noradrenaline-type granules. CONCLUSION: The lack of correlation between hormone output and granularity suggests that other factors determine secretory patterns in these tumours. PMID- 9876075 TI - Methods of repair and risk for reoperation in Swedish hernia surgery from 1992 to 1996. AB - BACKGROUND: Difficulties in obtaining and analysing outcome measures in hernia surgery may be an obstacle to necessary progress in non-specialized hospitals. Against this background a voluntary register was initiated in 1992 with the aim of describing and evaluating hernia surgery in participating units. METHODS: Prospective registration of all hernia operations carried out in participating hospitals was undertaken using identification codes specific for each individual. Repair technique, complications, day surgery, type of anaesthesia, and reoperation for recurrence were recorded. Actuarial analysis was used to determine the cumulative incidence of reoperation. Relative risk for reoperation was estimated by the Cox proportional hazards model. RESULTS: The number of participating hospitals and registered operations increased from eight and 1689 respectively in 1992 to 21 and 4056 in 1996. The use of mesh increased from 7 per cent of all operations in 1992 to 51 per cent in 1996. The proportion of operations done for recurrent hernia remained constant at 16-17 per cent throughout the 5-year study period. For all 12542 herniorrhaphies registered, the cumulative incidence of reoperation at 2 years was 3 (95 per cent confidence interval 3-4) per cent. Postoperative complications, recurrent hernia, direct hernia and absorbable suture were associated with increased risk of reoperation for recurrence. An increased incidence of reoperation, although not statistically significant, was noted for conventional open repairs (Bassini, McVay, Marcy and others) versus the Shouldice technique. CONCLUSION: In this prospective audit an increasing use of mesh was observed for open and laparoscopic surgery, especially for bilateral and recurrent hernia operations. Reoperation rates decreased significantly between 1992 and 1995. PMID- 9876076 TI - Water-soluble contrast study predicts the need for early surgery in adhesive small bowel obstruction. AB - BACKGROUND: The optimal period of conservative treatment for adhesive small bowel obstruction remains controversial. This study sought to determine whether a 24-h abdominal radiograph after oral Urografin is a reliable indicator for operation in patients with adhesive small bowel obstruction. METHODS: One hundred and sixty one patients who suffered from adhesive intestinal obstruction without clinical evidence of strangulation or gangrene underwent a Urografin study. Some 40 ml Urografin mixed with 40 ml distilled water was administered either orally or via a nasogastric tube to each patient. Serial plain abdominal radiographs were taken 4, 8, 16 and 24 h later. If an earlier plain radiograph showed that contrast medium had reached the ascending colon, subsequent radiographs were not taken. RESULTS: Contrast medium reached the colon within 24 h in 112 patients (70 per cent). These patients were all treated successfully with non-operative methods. Contrast medium was not observed in the colon within the first 24 h in 49 patients (30 per cent). Operation was performed in 47 of these patients and non operative treatment was given in two. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of Urografin reaching the colon within 24 h as an indicator for non-operative treatment were 98, 100, 99, 100 and 96 per cent respectively. CONCLUSION: All patients with evidence of Urografin reaching the colon within 24 h were treated successfully with non operative methods. The results of this prospective study suggest that patients with adhesive intestinal obstruction in whom contrast medium fails to reach the colon within 24 h should receive prompt surgical intervention. PMID- 9876077 TI - Advancement sleeve flaps for treatment of severe perianal Crohn's disease. AB - BACKGROUND: Traditionally proctectomy has been the treatment for severe, complex fistula in ano from Crohn's disease. However, based on the success of rectal advancement flaps in Crohn's disease, circumferential transanal sleeve advancement flaps (TSAFs) were proposed for this subgroup of patients with severe fistula. METHODS: From 1991 to 1995, 13 patients (12 women) with severe perianal Crohn's disease and multiple fistula tracts underwent a TSAF procedure. Data were collected retrospectively using a standard data sheet. RESULTS: There were no postoperative deaths or major morbidity. One year after surgery, the fistula had healed in eight of 13 patients (with three requiring additional surgery before healing). Of patients in whom the procedure failed, three underwent proctectomy for progression of disease and the other two had recurrence of a rectovaginal fistula 6 and 8 months after surgery. Of six variables evaluated (previous procedure, steroid use, steroid dosage, associated Crohn's disease, associated procedures and diverting stoma), only associated procedures were significantly related to a successful outcome (P=0.008). CONCLUSION: Some patients with severe perianal Crohn's fistula and a relatively normal rectum can be offered TSAFs. Even with successful outcome in eight of 13 patients, this may still be a viable option if the only alternative would be total proctocolectomy and a permanent stoma. PMID- 9876078 TI - Pathophysiology and treatment of anterior rectal mucosal prolapse syndrome. AB - BACKGROUND: The study was designed to investigate the clinical presentation and laboratory findings of anterior rectal mucosal prolapse (ARMP) and to assess the results of two therapeutic modalities. METHODS: Some 162 women with ARMP were assessed clinically and by defaecography and rectoanal manometry before and 1 year after one or two sessions of submucosal sclerotherapy or, in the case of recurrence, after transanal excision of the prolapsing mucosa. RESULTS: Almost all patients reported a combination of symptoms suggesting obstructive defaecation. At defaecography anterior rectocele and excessive perineal descent at straining were present in 78 and 72 per cent respectively. The size of coexisting anterior rectocele and the extent of perineal descent were significantly related to the duration of the disease (P< 0.001). One or, in the event of recurrence, two sessions of sclerotherapy led to an overall success rate of 51 per cent. Improvement after sclerotherapy was associated with partial recovery of anal tone and improvement of anal sampling and rectal sensation. Failure of sclerotherapy was related to rectocele of larger size (P< 0.001) and a longer perineal descent at straining (P< 0.001) than in patients with a successful outcome. Excision of the prolapsing mucosa resulted in symptomatic improvement in 42 of 47 patients and was associated with significant improvement of the defaecographic and manometric findings. CONCLUSION: ARMP is usually associated with anterior rectocele and excessive perineal descent. Submucosal sclerotherapy is successful in half of the cases, but only in the presence of a rather small anterior rectocele and short perineal descent. Failures after sclerotherapy can be treated by transanal excision of the prolapsing mucosa. PMID- 9876079 TI - Effect of a novel prosthetic anal neosphincter on human colonic blood flow. AB - BACKGROUND: An artificial anal sphincter has been developed which aimed to simulate the normal physiology of the anorectum. As a prelude to human implantation the present study reports the effect of inflation of this device on colonic perfusion in patients undergoing colectomy, which was assessed by using a laser Doppler scanner. METHODS: Eleven patients (median age 49.8 (range 24.3 78.7) years) were studied. Five patients had inflammatory bowel disease (IBD). The neosphincter was placed around the bowel and progressively inflated. The model was designed so that blood flow changes to the colon under the neosphincter would be reflected in the distal segment of the bowel, which could be scanned by the laser Doppler scanner. RESULTS: The blood flow in the colon distal to the device was significantly higher in patients with IBD (mean(s.e.m.) 288.6(71.9) versus 211.1(57.6) perfusion units; P < 0.001). The mean(s.e.m.) 'biological zero' value was 46(14) perfusion units. Blood flow distal to the neosphincter decreased progressively with increased sphincteric compression by 0.66 per cent per mmHg applied pressure in controls and 0.35 per cent per mmHg in patients with IBD (P < 0.05). CONCLUSION: These results suggest that at the planned operating occlusion pressure (less than 45 mmHg) this neosphincter should not put the vascularity of the human colon at risk. PMID- 9876080 TI - Prospective assessment of the hepatic perfusion index in patients with colorectal cancer. AB - BACKGROUND: This prospective study was designed to test the hypothesis that abnormal liver blood flow is related to poor prognosis in patients with colorectal cancer. METHODS: The hepatic perfusion index (HPI), measured by dynamic hepatic scintigraphy, was assessed in 202 patients with colorectal cancer. Assessment for overt hepatic metastasis included liver palpation at laparotomy and perioperative computed tomography (CT). Follow-up at a dedicated clinic included regular abdominal ultrasonography and CT. RESULTS: The HPI was abnormal (greater than 0.37) in 92 (88 per cent) of 105 patients with overt liver metastases. Of 89 patients with no evidence of overt metastases or residual tumour after primary resection, 52 had an abnormal and 37 a normal HPI. At a median follow-up of 39 (range 13-76) months, 25 of 38 patients with recurrence had an abnormal HPI. Some 31 of 45 patients who died had an abnormal HPI. The HPI predicted overall recurrence (P=0.04, log rank test). Multivariate analysis showed the HPI was independent of Dukes stage for predicting disease-free survival (P=0.04, relative risk 1.94 (95 per cent confidence interval (c.i.) 1.03 3.67)) but this just failed to attain significance for overall survival (P=0.055, relative hazard 1.88 (95 per cent c.i. 1.00-3.58)). CONCLUSION: The HPI predicts a poor outcome in patients with colorectal cancer and may be useful in patient selection for adjuvant chemotherapy. PMID- 9876081 TI - Ultrasonographically-guided fine-needle aspiration cytology in the diagnosis of colonic lesions. AB - BACKGROUND: The use of fine-needle aspiration cytology (FNAC) in the diagnosis of colonic lesions was investigated. METHODS: Some 22 patients (median age 71 years) with a colonic lesion identified on abdominal ultrasonography underwent ultrasonographically-guided FNAC using a 21-G needle. The sample was checked immediately by a cytopathologist for adequacy. RESULTS: Eighteen patients had colonic carcinoma; aspiration cytology detected malignant epithelial cells consistent with colonic carcinoma in 17 patients and severely dysplastic cells in one patient. The sensitivity and specificity of ultrasonographically-guided FNAC in the diagnosis of colonic carcinoma was 94 and 100 per cent respectively. The remaining four patients had a diagnosis of ileocaecal tuberculosis, ileocaecal Crohn's disease, and metastatic adenocarcinoma in the liver with no identifiable primary (two patients). One demonstrated granulomata, grew acid-fast bacilli and the patient was treated for tuberculosis. One had inflammatory cells and the patient was found to have Crohn's disease on histology. The remaining two patients had confirmed metastatic adenocarcinoma in the liver on aspiration cytology but suspected colonic lesions were found to be benign on cytological examination and no primary lesion was subsequently demonstrated. There were no complications of FNAC and patients complained of minimal discomfort. There has been no evidence of tumour recurrence with a median follow-up of 12 (range 1-25) months. CONCLUSION: Ultrasonographically-guided FNAC is a valid method for the diagnosis of colonic tumours. PMID- 9876082 TI - Quality of life in patients with benign anorectal disorders. AB - BACKGROUND: Benign proctological conditions are very common in Western civilization. However, to date quality of life in these patients has not been evaluated comprehensively. The aim of this study was to investigate whether the Gastrointestinal Quality of Life Index (GIQLI) is a useful instrument for measuring quality of life in these patients, and subsequently to establish baseline values for different anorectal disorders. METHODS: The questionnaire was completed by 325 consecutive patients (182 men; mean age 49 years) seen at the proctology outpatient clinic. For further analysis patients were classified into nine subgroups according to the primary diagnosis: group 1, haemorrhoids (n=96); group 2, anal fissure (n=38); group 3, fistula in ano (n=22); group 4, severe constipation (n=14); group 5, faecal incontinence (n=35); group 6, symptomatic anterior rectocele (n=12); group 7, perianal abscess (n=7); group 8, perianal thrombosis (n=7); and group 9, miscellaneous conditions (e.g. skin tags, anal papillomas, mild constipation, rectal polyps) (n=94). The GIQLI scores were compared between the subgroups. In addition the GIQLI scores of age-matched healthy controls, derived from data in the literature, were compared with the patients' scores. RESULTS: The mean GIQLI score for all patients was 113 (78.5 per cent of the maximum score of 144). Mean scores for the nine diagnostic subgroups were: group 1, 120; group 2, 104; group 3, 119; group 4, 94; group 5, 93; group 6, 112; group 7, 115; group 8, 129; and group 9, 117. Age-matched controls from a series published previously had a significantly higher GIQLI score compared with all patients (P < 0.0001). However, only the subgroups of patients with miscellaneous conditions, fissures, severe constipation and faecal incontinence had a significantly poorer quality of life than age-matched healthy individuals. CONCLUSION: The GIQLI is a valuable instrument for measuring quality of life in patients with benign anorectal disorders. Although certain diseases do not seem to affect quality of life profoundly, certain subgroups of patients, most notably those with incontinence and severe constipation, are extremely compromised. Severely constipated individuals exhibit the same poor quality of life as patients with faecal incontinence. PMID- 9876083 TI - Role of magnetic resonance angiography for assessment of abdominal aortic aneurysm before endoluminal repair. PMID- 9876084 TI - Cytogenetics and the surgeon: an invaluable tool in diagnosis, prognosis and counselling of patients with solid tumours. PMID- 9876085 TI - Venous insufficiency and perforating veins. PMID- 9876086 TI - Arteriovenous fistula using transposed basilic vein. PMID- 9876087 TI - Assessment of a scoring system for breast imaging. PMID- 9876088 TI - Assessment of a scoring system for breast imaging. PMID- 9876089 TI - Blood contacting surface of titanium: is it the metal or the water? PMID- 9876090 TI - Is ionic dialysance a valid parameter for quantification of dialysis efficiency? AB - The on-line measurement during hemodialysis of ionic dialysance provides an estimation of urea clearance with a good and already proven correlation. Some discrepancies remain controversial, and the influence of the dialyzer membrane is still being debated. Eighty-eight measurements of ionic dialysance (ID) were performed with a Diascan module (Hospal R&D, Int., Lyon, France), 51 with cellulosic membranes, and 37 with synthetic membranes, chosen according to their surface charges. The ID was compared to the urea clearance (UK) measured from the blood (n=16) and dialysate (n=88) sides. The ID is closely correlated (r=0.91) but significantly (p < 0.01) lower than the UK by 5% (ID/UK=0.95+/-0.06). The correlation is improved by a semilogarithmic regression analysis (r=0.93). Regarding the influence of the membrane charge, a slight difference is only evidenced for UK < 180 ml/min whereby ID is closer to the urea clearance for the charged membranes (ID/UK=0.98+/-0.05 for charged membranes versus 0.95+/-0.05 for noncharged membranes, p < 0.05). The discrepancy between ID and UK could be related with the difference in the blood distribution volume of urea and that of electrolytes. The good correlation provides the major argument for ID being used as a monitoring parameter of the delivered dialysis dose. Having integrated the discrepancy between ID and UK, prescription can be guided by ID for delivering the adequate normalized dialysis dose as defined by Kt/V. PMID- 9876091 TI - A newly developed on-line monitoring system for the determination of serum electrolytes and urea during hemodialysis. AB - Continuous monitoring of serum solute concentrations throughout a dialysis session could be beneficial for the prevention of intradialytic morbidity such as hypotension and for the confirmation of dialysis adequacy. In this paper, we introduce a new on-line monitoring system for use during hemodialysis sessions, a system developed for the continuous observation of serum biochemical data. Using this system, we also show the possibility of clinical applications. Intradialytic hypotension could be prevented by the observation of filtrate sodium concentration and the control of dialysate sodium concentration. The termination of a dialysis session could be decided by the observation of the filtrate urea nitrogen, on the basis of the Kt/V urea ratio. Because monitoring for other solutes such as calcium and creatinine is theoretically available, clinical applications will continue to increase in the future. PMID- 9876092 TI - Adsorption of complement, cytokines, and proteins by different dialysis membrane materials: evaluation by confocal laser scanning fluorescence microscopy. AB - The membranes tested in the present study were cellulose triacetate (CTA), polymethylmethacrylate (PMMA), and polyacrylonitrile (PAN). The adsorption by each membrane of albumin, IgG, C3a, interleukin-1beta (IL-1beta), interleukin-6 (IL-6), human neutrophil elastase (HNE), and tumor necrosis factor alpha (TNFalpha) was examined and semiquantitatively graded by confocal laser scanning fluorescence microscopy (CLSFM). After clinical use the dialyzers were treated with antibodies for these proteins and cytokines. Then the samples were incubated with fluorescein isothiocyanate-labeled anti-IgG antibody and observed by CLSFM. The changes in the blood levels of C3a and cytokines were also studied. In the CTA membrane, the adsorption of these substances, except for albumin and HNE, was less than in the synthetic membranes. The PAN membrane revealed the most abundant adsorption, especially for IL-1beta, IL-6, and TNFalpha. Although a marked elevation of C3a in the blood was observed in the CTA membrane, considerable adsorption was evident in the PMMA and the PAN membranes. Because the changes in the blood levels could be affected by membrane adsorption, both the blood levels and the adsorption of the biocompatibility parameters should be evaluated when membrane biocompatibility is discussed. PMID- 9876093 TI - Aprotinin reduces the expression of p-selectin on the surface of platelet and leukocyte-platelet conjugates. AB - P-Selectin, an adhesion molecule expressed on the surfaces of activated platelets and the vascular endothelium, mediates platelet binding to monocytes and neutrophils. Monocytes and neutrophils produce superoxide anion by activated platelets through p-selectin. Aprotinin, a serine protease inhibitor, inhibits plasmin to activate platelets during cardiopulmonary bypass (CPB). A total of 25 patients were studied to clarify the effects of aprotinin on p-selectin expression during CPB. Nine patients were not given aprotinin (control group), and 16 were given aprotinin of 2 million U in the priming solution (aprotinin group). The platelet count and soluble p-selectin in the plasma, p-selectin on the surface membranes of platelets, and leukocyte-platelet conjugate levels were measured during and after CPB. The platelet count was maintained well in the aprotinin group. The increases of soluble p-selectin in the plasma, platelet surface p-selectin, and leukocyte-platelet conjugates were less in the aprotinin group than in the control group (p < 0.05). In conclusion, aprotinin in patients undergoing CPB may reduce the early inflammatory reactions induced by p-selectin. PMID- 9876094 TI - The effects of serum from patients with acute liver failure on the growth and metabolism of Hep G2 cells. AB - In many bioartificial liver systems currently being designed and evaluated for use in fulminant hepatic failure, direct contact is required between the patient's blood and the liver cells in the device. The efficacy of such devices will be influenced by the interaction of fulminant hepatic failure (FHF) patient serum with the cells. We have found that FHF serum inhibits the growth rate and the synthesis of DNA, RNA, and protein; disturbs glutathione homeostasis; and induces morphological changes in cultured human Hep G2 cells. These interactions should influence the design of bioartificial liver devices based on proliferating cell lines and indicate the requirement to pretreat FHF patient plasma to reduce the toxin load. PMID- 9876095 TI - Efficacy of a bioreactor filled with porcine hepatocytes immobilized on nonwoven fabric for ex vivo direct hemoperfusion treatment of liver failure in pigs. AB - We developed a new bioreactor for a bioartificial liver filled with porcine hepatocytes immobilized on polyester nonwoven fabric (NWF) and in our previous study showed that this NWF bioreactor has promising in vitro efficiency. In the present study, we investigated the efficacy of the NWF bioreactor in a direct hemoperfusion experiment conducted to treat pigs with liver failure. Porcine hepatocytes were isolated from the whole liver of a Sangen strain pig. They were immobilized in a 200 ml column containing NWF via perfusion in a closed circuit for 24 h to prepare a NWF bioreactor. The following day an operative liver failure model was produced by creating a portocaval shunt and ligating the entire hepatoduodenal ligament in the porta hepatis. Perfusion treatment was initiated 4 h after operative induction of liver failure and continued for about 1 h. The pigs which underwent perfusion treatment showed significant improvements in survival and blood data, including ammonia, total bile acid, glucose, and prothrombin time, attributed to significant improvements in the post- as compared to the prebioreactor levels in the perfused blood of the treated pigs. These beneficial effects of the NWF bioreactor were based on its excellent composition which allows the accommodation of adequate numbers of hepatocytes and direct contact between hepatocytes and perfused blood. PMID- 9876096 TI - Therapeutic apheresis for septic patients with organ dysfunction: hemoperfusion using a polymyxin B immobilized column. AB - A prospective clinical study was performed to evaluate a new method of treatment of endotoxin shock, a column containing polystyrene fibers with covalently bound immobile polymyxin B. Direct hemoperfusion using the column removes circulating endotoxin by adsorption. All of the patients studied, 37 in the treatment group and 33 in the control group, had endotoxemia and failure of 1 or more organs. The perfusion was performed 1-7 times per patient, 2 h/session. The survival rate was significantly higher in the treatment group (54%) than in the controls (36.4%). The mean plasma endotoxin concentration was significantly lowered by the treatment from 83.7 pg/ml before perfusion to 56.4 pg/ml immediately after and 28.5 pg/ml the day after the treatment, and the posttreatment level was much lower in those who survived (mean, 18.8 pg/ml) compared to those who died (mean, 88 pg/ml). Various parameters of cardiac function also improved after the treatment. PMID- 9876097 TI - Induced cell trauma during in vitro perfusion: a comparison between two different perfusion systems. AB - The purpose of this study was to compare blood cell activation during in vitro long-term perfusion using 2 parallel in vitro extracorporeal membrane oxygenation (ECMO) systems. We compared two substantially different perfusion systems, an assistance respiratoire extra corporelle (AREC) system on one hand, containing an AREC pump, silicon tubing, and a hollow-fiber oxygenator, and a centrifugal pump system, on the other hand, containing a Biomedicus centrifugal pump, PVC tubing, and a membrane oxygenator. We measured the platelet count using an automated blood cell counter. Platelet activation was evaluated using a flow cytometric technique for the platelet membrane expression of glycoproteins and ELISA for the plasma concentration of beta-thromboglobulin (beta-TG), a platelet specific protein released into the blood upon platelet activation. The neutrophil count was assayed using an automated blood cell counter and the plasma concentration of cytokines using an ELISA kit. A significant difference between the two systems was observed in terms of the platelet membrane expression of glycoprotein (GP)Ib (p=0.0001) and GPIIb/IIIa (p=0.0037), indicating a lower degree of platelet activation in the AREC system. The concentration of neutrophils was significantly lower in the centrifugal system (p=0.002) compared to the AREC system. The neutrophil membrane expression of CD11b was significantly lower (p=0.0067) in the AREC system, indicating a lower degree of neutrophil activation compared to the centrifugal pump system. A significantly lower degree of hemolysis, as expressed by plasma hemoglobin, was observed in the AREC pump system (p=0.0491). In conclusion, lower degrees of the platelet membrane expression of GPIb and GPIIb/IIIa and of the neutrophil membrane expression of CD11b were observed in the AREC system, indicating a lower degree of platelet and neutrophil activation in this system. No significant difference between the two systems as to the plasma concentration of interleukin (IL)-1beta, IL-6, or IL-8 could be recorded. Further studies are warranted to specify the role of each individual component of the two systems. PMID- 9876098 TI - Effect of ultrafiltration during cardiopulmonary bypass for pediatric cardiac surgery. AB - The effect of ultrafiltration during cardiopulmonary bypass (CPB) was evaluated for correcting ventricular septal defects with associated pulmonary hypertension in patients less than 18 months old. Interleukin (IL)-6 and IL-8 concentrations in the blood, ultrafiltrate, and urine were measured. The blood IL-6 concentration increased to 128.4+/-20.2 pg/ml by the end of surgery, which is lower than the concentration seen in adult patients (273.1+/-48.2 pg/ml, p < 0.02). The blood IL-8 concentration was not significantly different than that of adults. The total amounts of excreted IL-6 in the ultrafiltrate and urine during CPB were 11.5+/-0.32 pg/kg and 0.32+/-0.07 pg/kg, respectively (p < 0.05). The total amounts of excreted IL-8 in the ultrafiltrate and urine were 4.64+/-0.69 pg/kg and 1.92+/-0.56 pg/kg, respectively (p < 0.05). No differences were seen in these values for excretion between children and adults. We conclude that ultrafiltration during CPB in pediatric patients is more effective in removing proinflammatory cytokines than in adults and more effective than renal filtration alone. PMID- 9876099 TI - Arterial baroreceptor afferent activity in nonpulsatile systemic circulation. AB - We directly measured aortic depressor nerve activity (ADNA) in rabbits to investigate changes in arterial baroreceptor afferent activity (ABAA) in response to nonpulsatile systemic circulation (NC) during total left heart bypass (TLHB). ADNA was measured before and 5, 20, 30, 40, 50, and 60 min after the start of TLHB while maintaining the mean aortic pressure (MAP) at preestablished values (n=10). At 5 min, the ADNA was significantly increased compared to its baseline value. At 30 min, it was significantly decreased compared to its value at 5 min and thereafter remained the same. The ADNA was then measured before and 5 and 30 min after the start of TLHB under increased MAP (n=10). The results showed that the percentage changes in ADNA during TLHB were significantly less than that observed when the MAP was increased before TLHB. We conclude that ABAA acutely adapts to NC 30 min after its initiation, but changes in ABAA are depressed under NC combined with increased MAP. PMID- 9876100 TI - Lixelle adsorbent to remove inflammatory cytokines. AB - A beta2-microglobulin (beta2M) selective adsorbent (Lixelle) for direct hemoperfusion has been used for the treatment of hemodialysis patients with the long-term complication of dialysis related amyloidosis (DRA), but there is no significant correlation between the serum level of beta2M and the occurrence of DRA. Inflammatory cytokines such as interleukin (IL)-1, IL-6, and tumor necrosis factor alpha (TNFalpha) are related to the development of DRA. We studied the adsorptive rates of cytokines in vitro using the Lixelle adsorbent. The adsorptive rates were 98.5% for IL-1beta 98.0% for interleukin-1 receptor antagonist (IL-1RA), 82.9% for IL-6, 99.9% for IL-8, 31.2% for TNFalpha, and 46.1% for soluble TNF receptor (sTNFr), respectively. As the molecular weights of cytokines increase, the adsorptive rates decrease. The Lixelle column adsorbed beta2M and various inflammatory cytokines as well. Therefore, the removal of both beta2M and inflammatory cytokines may play an important role in the treatment of DRA. PMID- 9876101 TI - Complement activation as a cause of transient hypotension during plasmapheresis. AB - Hypotension is one of the most common adverse effect of plasmapheresis (PP) and often is attributed to hypovolemia due to extracorporeal circulation and the vasovagal reflex. Complements are activated during PP, and the activated complements are strong anaphylatoxins and potent vasodilators. Therefore, we studied the relationship between the transient hypotension and the plasma levels of activated complements during and after PP in 8 sessions of 7 patients using the Plasmafro OP-08 as a plasma separator. Five of the patients underwent immunoadsorption PP using the IM-TR 350 or IM-PH 350 as the adsorption column. The other underwent double filtration PP using the Evaflux 4A as a second filter. In 4 of 8 sessions, patients experienced transient hypotension with significantly elevated plasma levels of activated complements C3a and C5a. In contrast, patients without hypotension showed no increases in C3a and C5a values during PP. In this report, we emphasize the critical role of activated complements for hypotension during PP. PMID- 9876103 TI - Estimation of the minimum pump speed to prevent regurgitation in the continuous flow left ventricular assist device: left ventricular drainage versus left atrial drainage. PMID- 9876102 TI - Views on the artificial liver. PMID- 9876104 TI - Leptin, a pleiotropic hormone: physiology, pharmacology, and strategies for discovery of leptin modulators. PMID- 9876105 TI - Novel analogues of arachidonylethanolamide (anandamide): affinities for the CB1 and CB2 cannabinoid receptors and metabolic stability. AB - Several analogues of the endogenous cannabinoid receptor ligand arachidonylethanolamide (anandamide) were synthesized and evaluated in order to study (a) the structural requirements for high-affinity binding to the CB1 and CB2 cannabinoid receptors and (b) their hydrolytic stability toward anandamide amidase. The series reported here was aimed at exploring structure-activity relationships (SAR) primarily with regard to stereoelectronic requirements of ethanolamido headgroup for interaction with the cannabinoid receptor active site. Receptor affinities, reported as Ki values, were obtained by a standard receptor binding assay using [3H]CP-55,940 as the radioligand, while stability toward the amidase was evaluated by comparing the Ki of each analogue in the presence and absence of phenylmethanesulfonyl fluoride (PMSF), a serine protease blocker and inhibitor of anandamide amidase. Introduction of a methyl group in the 1'- and 2' positions or substitution of the ethanolamido headgroup with a butylamido group gave analogues with vastly improved biochemical stability. This is accomplished in some cases with increased receptor affinity. Conversely, oxazolyl and methyloxazolyl headgroups led to low-affinity analogues. Substitution of the hydroxyl group with electronegative substituents such as fluoro, chloro, allyl, and propargyl groups significantly increased receptor affinity but did not influence the biochemical stability. The 2'-chloro analogue of anandamide was found to have the highest affinity for CB1. Additionally, reversing the positions of the carbonyl and NH in the amido group produces retro-anandamides possessing considerably higher metabolic stability. Replacement of the arachidonyl tail with oleyl or linoleyl results in analogues with low affinities for both receptors. All of the analogues in this study showed high selectivity for the CB1 receptor over the peripheral CB2 receptor. The most potent analogues were tested for their ability to stimulate the binding of [35S]GTPgammaS to G-proteins and were shown to be potent cannabimimetic agonists. The results are discussed in terms of pharmacophoric features affecting receptor affinity and enzymatic stability. PMID- 9876106 TI - Development of dual-acting agents for thromboxane receptor antagonism and thromboxane synthase inhibition. 3. Synthesis and biological activities of oxazolecarboxamide-substituted omega-phenyl-omega-(3-pyridyl)alkenoic acid derivatives and related compounds. AB - A novel series of oxazolecarboxamide-substituted omega-phenyl-omega-(3 pyridyl)alkenoic acid derivatives was discovered as potent dual-acting agents to block the TXA2 receptor and to inhibit the thromboxane synthase (TRA/TSI). Synthesis, structure-activity relationship (SAR), and in vitro and in vivo pharmacology of this series of compounds are described. Modification of the series revolved around the oxazole moiety to increase the hydrophilicity of the compounds and to correlate the biological activity with lipophilicity of the compounds. The most potent in the series was (E)-7-[4-[4-[[(4 cyclohexylbutyl)amino]carbonyl]-2-oxazolyl] phenyl]-7 -(3-pyridyl)hept-6-enoic acid (14) with Kd = 9.9 +/- 0.4 nM for the thromboxane receptor antagonism and IC50 = 55.0 +/- 17.9 nM for thromboxane synthase inhibition. The compound 14 was a selective TRA/TSI which exhibited desirable characteristics for oral activity, "shunt" effect to elevate PGI2 level, and absence of agonist activity. PMID- 9876107 TI - The 16,17-double bond is needed for irreversible inhibition of human cytochrome p45017alpha by abiraterone (17-(3-pyridyl)androsta-5, 16-dien-3beta-ol) and related steroidal inhibitors. AB - Abiraterone (17-(3-pyridyl)androsta-5,16-dien-3beta-ol, 1) is a potent inhibitor (IC50 4 nM for hydroxylase) of human cytochrome P45017alpha. To assist in studies of the role of the 16,17-double bond in its mechanism of action, the novel 17alpha-(4-pyridyl)androst-5-en-3beta-ol (5) and 17beta-(3-pyridyl)-16,17alpha epoxy-5alpha-androst-3beta-ol (6) were synthesized. 3beta-Acetoxyetienic acid was converted in three steps into 5 via photolysis of the thiohydroxamic ester 8. Oxidation of an appropriate 16,17-unsaturated precursor (21) with CrO3-pyridine afforded the acetate (23) of 6. Inhibition of the enzyme by 1, the similarly potent 5,6-reduced analogue 19 (IC50 5 nM), and the 4, 16-dien-3-one 26 (IC50 3 nM) and by the less potent (IC50 13 nM) 3,5, 16-triene 25 is slow to occur but is enhanced by preincubation of the inhibitor with the enzyme. Inhibition following preincubation with these compounds is not lessened by dialysis for 24 h, implying irreversible binding to the enzyme. In contrast under these conditions the still potent (IC50 27 nM) 17alpha-(4-pyridyl)androst-5-en-3beta-ol (5) showed partial reversal after 5 h of dialysis and complete reversal of inhibition after 24 h. This behavior was also shown by the less potent 16,17-reduced 3-pyridyl compounds 3 and 24. Further, in contrast to the compounds (1, 19, 25, 26) with the 16,17 double bond, the inhibition of the enzymic reaction was not enhanced by preincubation either with 5 or with the 17beta-pyridyl analogues 3, 4, and 24 which also lack this structural feature. The results show that the 16,17-double bond is necessary for irreversible binding of these pyridyl steroids to cytochrome P45017alpha. However oxidation to an epoxide is probably not involved since epoxide 6 was only a moderately potent inhibitor (IC50 260 nM). PMID- 9876108 TI - Evaluation of dynamic polar molecular surface area as predictor of drug absorption: comparison with other computational and experimental predictors. AB - The relationship between various molecular descriptors and transport of drugs across the intestinal epithelium was evaluated. The monolayer permeability (Pc) of human intestinal Caco-2 cells to a series of nine beta-receptor-blocking agents was investigated in vitro. The dynamic polar molecular surface area (PSAd) of the compounds was calculated from all low-energy conformations identified in molecular mechanics calculations in vacuum and in simulated chloroform and water environments. For most of the investigated drugs, the effects of the different environments on PSAd were small. The exception was H 216/44, which is a large flexible compound containing several functional groups capable of hydrogen bonding (PSAd,chloroform = 70.8 A2 and PSAd,water = 116.6 A2). The relationship between Pc and PSAd was stronger than those between Pc and the calculated octanol/water distribution coefficients (log Dcalc) or the experimentally determined immobilized liposome chromatography (ILC) retention. Pc values for two new practolol analogues and H 216/44 were predicted from the structure permeability relationships of a subset of the nine compounds and compared with experimental values. The Pc values of the two practolol analogues were predicted well from both PSAd calculations and ILC retention studies. The Pc value of H 216/44 was reasonably well-predicted only from the PSAd of conformations preferred in vacuum and in water. The other descriptors overestimated the Pc of H 216/44 100-500-fold. PMID- 9876110 TI - 7-[3-(1-piperidinyl)propoxy]chromenones as potential atypical antipsychotics. 2. Pharmacological profile of 7-[3-[4-(6-fluoro-1, 2-benzisoxazol-3-yl)-piperidin-1 yl]propoxy]-3-(hydroxymeth yl)chromen -4-one (abaperidone, FI-8602). AB - A series of novel 7-[3-(1-piperidinyl)propoxy]chromenones was synthesized and tested as potential antipsychotics in several in vitro and in vivo assays. The compounds possessed good affinity for D2 receptors, together with a greater affinity for 5-HT2 receptors, a profile which has been proposed as a model for atypical antipsychotics. Several agents also displayed a high potency in the climbing mice assay on oral administration, suggesting a potent antipsychotic effect as compared to reference standards. Compound 23 was selected for further pharmacological evaluation. Induction of catalepsy and inhibition of stereotypies weaker than standards, along with a lower increase in serum prolactin levels, were indicative of a potential atypical profile for this compound. From these results, 7-[3-[4-(6-fluoro-1, 2-benzisoxazol-3-yl)piperidin-1-yl]propoxy]-3 (hydroxymethyl )chromen- 4-one (23, abaperidone) has been proposed for clinical evaluation in humans as a potential atypical antipsychotic. PMID- 9876109 TI - New 1,4-dihydropyridines conjugated to furoxanyl moieties, endowed with both nitric oxide-like and calcium channel antagonist vasodilator activities. AB - A series of 4-phenyl-1,4-dihydropyridines substituted at the ortho and meta positions of the phenyl ring with NO-donating furoxan moieties and their non-NO releasing furazan analogues were synthesized and pharmacologically characterized. The vasodilator activities of these compounds were evaluated on rat aorta and expressed as EC50 values or as EC50iGC values when obtained in the presence of inhibitors of guanylate cyclase methylene blue (MB) and 1H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one (ODQ). Affinities to 1, 4-DHP receptors on Ca2+ channels, expressed as IC50 values, were determined through displacement experiments of [3H]nitrendipine on rat cortex homogenates. A linear correlation between IC50 and EC50 values was found for compounds unable to release NO. EC50calcd values for derivatives containing NO-donor moieties, expression of the Ca2+-blocking component of their vasodilator activity, were interpolated on this linear regression. They showed a good correspondence with EC50iGC values determined in the presence of soluble guanylate cyclase inhibitors. Analysis of EC50iGC/EC50 ratios provided a useful tool to distinguish well-balanced hybrids from derivatives biased toward Ca2+-blocking or NO-dependent vasodilator activity. A detrimental effect on affinity to the 1, 4-DHP receptor, due to substitution at the ortho and meta positions of the 4-phenyl ring, was observed. SAR to explain this effect is proposed. PMID- 9876111 TI - Homocamptothecins: synthesis and antitumor activity of novel E-ring-modified camptothecin analogues. AB - Homocamptothecin (hCPT), a camptothecin (CPT) analogue with a seven membered beta hydroxylactone which combines enhanced plasma stability and potent topoisomerase I (Topo I)-mediated activity, is an attractive template for the elaboration of new anticancer agents. Like CPT, hCPT carries an asymmetric tertiary alcohol and displays stereoselective inhibition of Topo I. The preparation and biological screening of racemic hCPT analogues are described. The 10 hCPTs tested were better Topo I inhibitors than CPT. Fluorinated hCPTs 23c, d,f,g were found to have potent cytotoxic activity on A427 and PC-3 tumor cell lines. Their cytotoxicity remained high on the K562adr and MCF7mdr cell lines, which overexpress a functionally active P-glycoprotein. Fluorinated hCPTs were more efficacious in vivo than CPT on HT-29 xenografts. In this model, a tumor growth delay of 25 days was reached with hCPT 23g at a daily dose of 0.32 mg/kg, compared to 4 days with CPT at 0.625 mg/kg. Thus difluorinated hCPT 23g warrants further investigation as a novel Topo I inhibitor with high cytotoxicity toward tumor cells and promising in vivo efficacy. PMID- 9876112 TI - 4-(trans-4-Methylcyclohexyl)-4-oxobutyric acid (JTT-608). A new class of antidiabetic agent. AB - During an investigation of drugs for improving the beta-cell response to glucose, we found that 4-cyclohexyl-4-oxobutyric acid selectively improved glucose stimulated insulin release and glucose tolerance in both normal and diabetic rats. A series of 4-cycloalkyl-4-oxobutyric acids and related compounds were synthesized and evaluated for their effects on the glucose tolerance test and fasting euglycemia. This study elucidated the structural requirements for drug activity and determined that the optimum compound was 4-(trans-4 methylcyclohexyl)-4-oxobutyric acid 7 (JTT-608). This compound improved glucose tolerance from an oral dose of 3 mg/kg and did not change fasting euglycemia even at an oral dose of 30 mg/kg. Selective improvement of glucose-induced insulin secretion was observed in studies using neonatal streptozotocin rats (nSTZ rats) and perfused pancreases isolated from nSTZ rats. PMID- 9876113 TI - Synthesis and antitumor evaluation of 2,5-disubstituted-indazolo[4, 3 gh]isoquinolin-6(2H)-ones (9-aza-anthrapyrazoles). AB - The synthesis and antitumor evaluation of 2, 5-disubstituted-indazolo[4,3 gh]isoquinolin-6(2H)-ones (9-aza-APs) are described. The key intermediates in the synthesis are benz[g]isoquinoline-5,10-diones which are substituted at positions 6 and 9 with groups of different nucleofugacity for SNAr displacements. The initial displacement of fluoride by a substituted hydrazine leads to the pyrazole analogues. Substitution of the remaining leaving group by an amine or BOC protected amines leads to the 9-aza-APs 12. These analogues were converted into their maleate or hydrochloride salts 13. In two cases, namely, 13x and 13z, sidearm buildup was also employed in the synthetic pathway. In vitro evaluation of 9-aza-APs against the human colon tumor cell line LoVo uncovered for most of the compounds a cytotoxic potency lower than that of DuP-941 or mitoxantrone and comparable to that of doxorubicin. Only analogues 13c, 13n, and 13ff were as cytotoxic as DuP-941. Interestingly, while DuP-941 was highly cross-resistant in the LoVo cell line resistant to doxorubicin (LoVo/Dx), the 9-aza-APs carrying a distal lipophilic tertiary amine moiety in both chains were capable of overcoming the MDR resistance induced in this cell line. The 9-aza-APs show outstanding in vivo antitumor activity against both systemic P388 murine leukemia and MX-1 human mammary carcinoma transplanted in nude mice. At their optimal dosages, congeners 13a-c, 13f, 13n, 13q, 13x, and 13dd were highly effective against P388 leukemia with T/C% of 200-381, while the T/C% value of DuP-941 was 147. In the MX-1 tumor model, 24 compounds elicited percentages of tumor weight inhibitions (TWI) ranging from 50% to 99%. Congeners 13d, 13k, 13l, 13x, 13z, and 13ee emerged as the most effective ones, with TWI% 96, simliar to that of DuP-941 (TWI% = 95). On the basis of their efficacy profile in additional experimental tumors and lack of cardiotoxicity in preclinical models, two congeners have surfaced as potential clinical candidates. PMID- 9876114 TI - Structural and functional analyses of benzamidine-based inhibitors in complex with trypsin: implications for the inhibition of factor Xa, tPA, and urokinase. AB - The trypsin-like serine proteinase superfamily contains a number of potential therapeutic targets, many of which are unsuitable for routine X-ray crystallographic studies. We have cocrystallized a selection of benzamidine-based inhibitors with bovine trypsin and solved their structures to a resolution of up to 1.7 A. Despite similar chemical formulas, the inhibitors exhibit a range of diverse binding modes that reflect their inhibitory spectra against the serine proteinases trypsin, thrombin, factor Xa, tissue-type plasminogen activator (tPA) and urokinase (uPA). In contrast to the compact folded conformations of thrombin inhibitors which allow optimal binding in the well-defined hydrophobic S2/S4 pocket of thrombin, those effective against factor Xa exhibit an extended conformation that allows occupation of the S3/S4 region, where hydrophobic and electrostatic interactions can stabilize the conformation. One group of inhibitors containing an N-terminal 2,4, 6-triisopropylphenylsulfonyl (TIPPS) moiety show little or no penetration into the S3/S4 subsites of trypsin. These latter sites are occluded in uPA, explaining why this class of compounds is effective against uPA. Despite presenting an extensive hydrophobic surface toward the solvent, the Ki values for TIPPS-containing compounds against trypsin is in the range 10(-7) to 10(-8) M. Comparison of the binding of a bis-benzamidine inhibitor in trypsin and tPA indicate that a shift in potency can be induced by relatively minor changes in binding mode. Implications for the inhibition of these proteinases are discussed. PMID- 9876115 TI - Structure-activity relationships for 1-phenylbenzimidazoles as selective ATP site inhibitors of the platelet-derived growth factor receptor. AB - 1-Phenylbenzimidazoles are shown to be a new class of ATP-site inhibitors of the platelet-derived growth factor receptor (PDGFR). Structure-activity relationships (SARs) are narrow, with closely related heterocycles being inactive. A systematic study of substituted 1-phenylbenzimidazoles showed clear SARs. Substituents at the 4'- and 3'-positions of the phenyl ring are tolerated but do not significantly improve activity, while substituents at the 2'-position abolish it. Substituents in the 2-, 4-, and 7-positions of the benzimidazole ring (with the exception of 4-OH) also abolish activity. Most substituents at the 5- and 6 positions maintain or increase activity, with the 5-OH, 5-OMe, 5-COMe, and 5 CO2Me analogues being >10-fold more potent than the parent 1-phenylbenzimidazole. The 5-OMe analogue was both the most potent inhibitor, and showed the highest selectivity (50-fold) between PDGFR and FGFR isolated enzymes, and also a moderately effective inhibitor (IC50 = 1.9 microM) of PDGF-stimulated PDGFR autophosphorylation in rat aorta smooth muscle cells. PMID- 9876116 TI - NIEHS workshop: unique marine/freshwater models for environmental health research. PMID- 9876117 TI - Spheres of influence: no POPS. Persistent organic pollutants. PMID- 9876118 TI - Biomolecular electrostatics with the linearized Poisson-Boltzmann equation. AB - Electrostatics plays a key role in many biological processes. The Poisson Boltzmann equation (PBE) and its linearized form (LPBE) allow prediction of electrostatic effects for biomolecular systems. The discrepancies between the solutions of the PBE and those of the LPBE are well known for systems with a simple geometry, but much less for biomolecular systems. Results for high charge density systems show that there are limitations to the applicability of the LPBE at low ionic strength and, to a lesser extent, at higher ionic strength. For systems with a simple geometry, the onset of nonlinear effects has been shown to be governed by the ratio of the electric field over the Debye screening constant. This ratio is used in the present work to correct the LPBE results to reproduce fairly accurately those obtained from the PBE for systems with a simple geometry. Since the correction does not involve any geometrical parameter, it can be easily applied to real biomolecular systems. The error on the potential for the LPBE (compared to the PBE) spans few kT/q for the systems studied here and is greatly reduced by the correction. This allows for a more accurate evaluation of the electrostatic free energy of the systems. PMID- 9876119 TI - Brownian dynamics simulations of interactions between aldolase and G- or F-actin. AB - Compartmentation of proteins in cells is important to proper cell function. Interactions of F-actin and glycolytic enzymes is one mechanism by which glycolytic enzymes can compartment. Brownian dynamics (BD) simulations of the binding of the muscle form of the glycolytic enzyme fructose-1,6-bisphosphate aldolase (aldolase) to F- or G-actin provide first-encounter snapshots of these interactions. Using x-ray structures of aldolase, G-actin, and three-dimensional models of F-actin, the electrostatic potential about each protein was predicted by solving the linearized Poisson-Boltzmann equation for use in BD simulations. The BD simulations provided solution complexes of aldolase with F- or G-actin. All complexes demonstrate the close contacts between oppositely charged regions of the protein surfaces. Positively charged surface regions of aldolase (residues Lys 13, 27, 288, 293, and 341 and Arg 257) are attracted to the negatively charged amino terminus (Asp 1 and Glu 2 and 4) and other patches (Asp 24, 25, and 363 and Glu 361, 364, 99, and 100) of actin subunits. According to BD results, the most important factor for aldolase binding to actin is the quaternary structure of aldolase and actin. Two pairs of adjacent aldolase subunits greatly add to the positive electrostatic potential of each other creating a region of attraction for the negatively charged subdomain 1 of the actin subunit that is exposed to solvent in the quaternary F-actin structure. PMID- 9876120 TI - Free energy determinants of binding the rRNA substrate and small ligands to ricin A-chain. AB - A continuum model is provided of the free energy terms that contribute to the molecular association of ricin A-chain (RTA) with the rRNA substrate and several small ligands. The model for RTA interactions with the RNA was taken from a previously proposed complex containing a 29-mer oligonucleotide hairpin (. Proteins 27:80-95), and models for the ligands were constructed from x-ray crystallographic structures. The calculated absolute free energies of complex formation for the RTA-RNA assembly and several single-residue substitutions are in good agreement with experimental data, given the approximations of evaluating the strain energy and conformational entropy. The free energy terms were found to resemble those of protein-protein complexes, with the net unfavorable electrostatic contribution offset by the favorable nonspecific hydrophobic effect. Decomposition of the RTA-RNA binding free energy into individual contributions revealed the electrostatic "hot" spots arising from charge-charge complementarity of the interfacial arginines with the RNA phosphate backbone. Base interactions of the GAGA loop structure dominate the hydrophobic complementarity. A linear-scaling model was parametrized for evaluating the binding of small ligands against the rRNA substrate and illustrates the free energy determinant required for designing specific RTA inhibitors. PMID- 9876121 TI - Alamethicin helices in a bilayer and in solution: molecular dynamics simulations. AB - Alamethicin is an alpha-helical channel-forming peptide, which inserts into lipid bilayers in a voltage-dependent, asymmetrical fashion. Nanosecond molecular dynamics simulations have been used to compare alamethicin conformation and dynamics in three different environments: 1) in water; 2) in methanol; and 3) inserted into a lipid (palmitoyl-oleoyl-phosphatidylcholine) bilayer to form a transmembrane helix. In the bilayer and in methanol, there was little change (Calpha RMSD approximately 0.2 nm over 2 ns and 1 ns) from the initial helical conformation of the peptide. In water there were substantial changes (Calpha RMSD approximately 0.4 nm over 1 ns), especially in the C-terminal segment of the peptide, which lost its alpha-helical conformation. In the bilayer and in methanol, the alamethicin molecule underwent hinge-bending motion about its central Gly-X-X-Pro sequence motif. Analysis of H-bonding interactions revealed that the polar C-terminal side chains of alamethicin provided an "anchor" to the bilayer/water interface via formation of multiple H-bonds that persisted throughout the simulation. This explains why the preferred mode of helix insertion into the bilayer is N-terminal, which is believed to underlie the asymmetry of voltage activation of alamethicin channels. PMID- 9876123 TI - Comparison of the crystal and solution structures of two RNA oligonucleotides. AB - Until recently, there were no examples of RNAs whose structures had been determined by both NMR and x-ray crystallography, and thus there was no experimental basis for assessing the accuracy of RNA solution structures. A comparison of the solution and the crystal structures of two RNAs is presented, which demonstrates that NMR can produce solution structures that resemble crystal structures and thus validates the application to RNA of a methodology developed initially for the determination of protein conformations. Models for RNA solution structures are appreciably affected by the parameters used for their refinement that describe intramolecular interactions. For the RNAs of interest here, the more realistic those parameters, the greater the similarity between solution structures and crystal structures. PMID- 9876122 TI - Molecular dynamics simulations of solvated yeast tRNA(Asp). AB - Transfer RNA molecules are involved in a variety of biological processes, implying complex recognition events with proteins and other RNAs. From a structural point of view, tRNAs constitute a reference system for studying RNA folding and architecture. A deeper understanding of their structural and functional properties will derive from our ability to model accurately their dynamical behavior. We present the first dynamical model of a fully neutralized and solvated tRNA molecule over a 500-ps time scale. Starting from the crystallographic structure of yeast tRNA(Asp), the 75-nucleotide molecule was modeled with 8055 water molecules and 74 NH4+ counterions, using the AMBER4.1 program and the particle mesh Ewald (PME) method for the treatment of long-range electrostatic interactions. The calculations led to a dynamically stable model of the tRNA molecule. During the simulation, all secondary and tertiary base pairs are maintained while a certain lability of base triples in the tRNA core is observed. This lability was interpreted as resulting from intrinsic factors associated with the "weaker" hydrogen bonding patterns seen in these base triples and from an altered ionic environment of the tRNA molecule. Calculated thermal factors are used to compare the dynamics of the tRNA in solution and in the crystal. The present molecular dynamics simulation of a complex and highly charged nucleic acid molecule attests to the fact that simulation methods are now able to investigate not only the dynamics of proteins, but also that of large RNA molecules. Thus they also provide a basis for further investigations on the structural and functional effects of chemical and posttranscriptionally modified nucleotides as well as on ionic environmental effects. PMID- 9876124 TI - Direct effects of phosphorylation on the preferred backbone conformation of peptides: a nuclear magnetic resonance study. AB - Control of protein activity by phosphorylation appears to work principally by inducing conformational change, but the mechanisms so far reported are dependent on the structural context in which phosphorylation occurs. As the activity of many small peptides is also regulated by phosphorylation, we decided to investigate possible direct consequences of this on the preferred backbone conformation. We have performed 1H nuclear magnetic resonance (NMR) experiments with short model peptides of the pattern Gly-Ser-Xaa-Ser, where Xaa represents Ser, Thr, or Tyr in either phosphorylated or unphosphorylated form and with either free or blocked amino and carboxy termini. The chemical shifts of amide protons and the 3JNH-Halpha coupling constants were estimated from one dimensional and two-dimensional scalar correlated spectroscopy (COSY) spectra at different pH values. The results clearly indicate a direct structural effect of serine and threonine phosphorylation on the preferred backbone dihedrals independent of the presence of charged groups in the surrounding sequence. Tyrosine phosphorylation does not induce such a charge-independent effect. Additionally, experiments with p-fluoro- and p-nitro-phenylalanine-containing peptides showed that the mere presence of an electronegative group on the aromatic ring of tyrosine does not produce direct structural effects. In the case of serine and threonine phosphorylation a strong dependence of the conformational shift on the protonation level of the phosphoryl group could be observed, showing that phosphorylation induces the strongest effect in its dianionic, i.e., physiological, form. The data reveal a hitherto unknown mechanism that may be added to the repertoire of conformational control of peptides and proteins by phosphorylation. PMID- 9876125 TI - Cysteines beta93 and beta112 as probes of conformational and functional events at the human hemoglobin subunit interfaces. AB - Three variants of tetrameric human hemoglobin, with changes at the alpha1beta2/alpha2beta1-interface, at the alpha1beta1/alpha2beta2-interface, and at both interfaces, have been constructed. At alpha1beta2/alpha2beta1-interface the beta93 cysteine was replaced by alanine (betaC93A), and at the alpha1beta1/alpha2beta2-interface the beta112 cysteine was replaced by glycine (betaC112G). The alpha1beta2 interface variant, betaC93A, and the alpha1beta1/alpha1beta2 double mutant, beta(C93A+C112G), were crystallized in the T-state, and the structures determined at 2. 0 and 1.8 A resolution, respectively. A comparison of the structures with that of natural hemoglobin A shows the absence of detectable changes in the tertiary folding of the protein or in the T-state quaternary assembly. At the beta112 site, the void left by the removal of the cysteine side chain is filled by a water molecule, and the functional characteristics of betaC112G are essentially those of human hemoglobin A. At the beta93 site, water molecules do not replace the cysteine side chain, and the alanine substitution increases the conformational freedom of beta146His, weakening the important interaction of this residue with beta94Asp. As a result, when Cl- is present in the solution, at a concentration 100 mM, the Bohr effect of the two mutants carrying the beta93Cys-->Ala substitution, betaC93A and beta(C93A+C112G), is significantly modified being practically absent below pH 7.4. Based on the crystallographic data, we attribute these effects to the competition between beta94Asp and Cl- in the salt link with beta146His in T-state hemoglobin. These results point to an interplay between the betaHis146-betaAsp94 salt bridge and the Cl- in solution regulated by the Cys present at position beta93, indicating yet another role of beta93 Cys in the regulation of hemoglobin function. PMID- 9876126 TI - Three-dimensional imaging of nerve tissue by x-ray phase-contrast microtomography. AB - We show that promising information about the three-dimensional (3D) structure of a peripheral nerve can be obtained by x-ray phase-contrast microtomography (p microCT; Beckmann, F., U. Bonse, F. Busch, and O. Gunnewig, 1997. J. Comp. Assist. Tomogr. 21:539-553). P-microCT measures electronic charge density, which for most substances is proportional to mass density in fairly good approximation. The true point-by-point variation of density is thus determined in 3D at presently 1 mg/cm3 standard error (SE). The intracranial part of the rat trigeminal nerve analyzed for the presence of early schwannoma "microtumors" displayed a detailed density structure on p-microCT density maps. The average density of brain and nerve tissue was measured to range from 0.990 to 0.994 g/cm3 and from 1.020 to 1.035 g/cm3, respectively. The brain-nerve interface was well delineated. Within the nerve tissue, a pattern of nerve fibers could be seen that followed the nerve axis and contrasted against the bulk by 7 to 10 mg/cm3 density modulation. Based on the fact that regions of tumor growth have an increased number density of cell nuclei, and hence of the higher z element phosphorus, it may become possible to detect very early neural "microtumors" through increases of average density on the order of 10 to 15 mg/cm3 by using this method. PMID- 9876127 TI - Detection of single mammalian cells by high-resolution magnetic resonance imaging. AB - This study reports the detection of single mammalian cells, specifically T cells (T lymphocytes) labeled with dextran-coated superparamagnetic iron oxide particles, using magnetic resonance microscopy. Size amplification due to sequestration of the superparamagnetic particles in vacuoles enhances contrast in localized areas in high-resolution magnetic resonance imaging. Magnetic resonance images of samples containing differing concentrations of T cells embedded in 3% gelatin show a number of dark regions due to the superparamagnetic iron oxide particles, consistent with the number predicted by transmission electron microscopy. Colabeling of T cell samples with a fluorescent dye leads to strong correlations between magnetic resonance and fluorescence microscopic images, showing the presence of the superparamagnetic iron oxide particles at the cell site. This result lays the foundation for our approach to tracking the movement of a specific cell type in live animals and humans. PMID- 9876128 TI - Hydrodynamics of micropipette aspiration. AB - The dynamics of human neutrophils during micropipette aspiration are frequently analyzed by approximating these cells as simple slippery droplets of viscous fluid. Here, we present computations that reveal the detailed predictions of the simplest and most idealized case of such a scheme; namely, the case where the fluid of the droplet is homogeneous and Newtonian, and the surface tension of the droplet is constant. We have investigated the behavior of this model as a function of surface tension, droplet radius, viscosity, aspiration pressure, and pipette radius. In addition, we have tabulated a dimensionless factor, M, which can be utilized to calculate the apparent viscosity of the slippery droplet. Computations were carried out using a low Reynolds number hydrodynamics transport code based on the finite-element method. Although idealized and simplistic, we find that the slippery droplet model predicts many observed features of neutrophil aspiration. However, there are certain features that are not observed in neutrophils. In particular, the model predicts dilation of the membrane past the point of being continuous, as well as a reentrant jet at high aspiration pressures. PMID- 9876130 TI - Long-time dynamics of Met-enkephalin: comparison of theory with Brownian dynamics simulations. AB - A recent theory for the long time dynamics of flexible chain molecules is applied for the first time to a peptide of biological importance, the neurotransmitter met-enkephalin. The dynamics of met-enkephalin is considerably more complicated than that of the previously studied glycine oligomers; met-enkephalin contains the interesting motions of phenyl groups and of side chains relative to the backbone, motions that are present in general flexible peptides. The theory extends the generalized Rouse (GR) model used to study the dynamics of polymers by providing a systematic procedure for including the contributions from the memory function matrices neglected in the GR theory. The new method describes the dynamics by time correlation functions instead of individual trajectories. These correlation functions are analytically expressed in terms of a set of equilibrium averages and the eigenvalues and eigenfunctions of the diffusion operator. The predictions of the theory are compared with Brownian dynamics (BD) simulations, so that both theory and simulation use identical potential functions and solvent models. The theory thus contains no adjustable parameters. Inclusion of the memory function contributions profoundly affects the dynamics. The theory produces very good agreement with the BD simulations for the global motions of met-enkephalin. It also correctly predicts the long-time relaxation rate for local motions. PMID- 9876129 TI - Physical origin of selectivity in ionic channels of biological membranes. AB - This paper shows that the selectivity properties of monovalent cation channels found in biological membranes can originate simply from geometrical properties of the inner core of the channel without any critical contribution from electrostatic interactions between the permeating ions and charged or polar groups. By using well-known techniques of statistical mechanics, such as the Langevin equations and Kramer theory of reaction rates, a theoretical equation is provided relating the permeability ratio PB/PA between ions A and B to simple physical properties, such as channel geometry, thermodynamics of ion hydration, and electrostatic interactions between the ion and charged (or polar) groups. Diffusive corrections and recrossing rates are also considered and evaluated. It is shown that the selectivity found in usual K+, gramicidin, Na+, cyclic nucleotide gated, and end plate channels can be explained also in the absence of any charged or polar group. If these groups are present, they significantly change the permeability ratio only if the ion at the selectivity filter is in van der Waals contact with them, otherwise these groups simply affect the channel conductance, lowering the free energy barrier of the same amount for the two ions, thus explaining why single channel conductance, as it is experimentally observed, can be very different in channels sharing the same selectivity sequence. The proposed theory also provides an estimate of channel minimum radius for K+, gramicidin, Na+, and cyclic nucleotide gated channels. PMID- 9876131 TI - Monte Carlo simulations of beta-hairpin folding at constant temperature. AB - Monte Carlo simulations were applied to beta-hairpin folding of a valine-based peptide. Two valine residues in the middle of the peptide were substituted with glycine, to serve as turn residues. Unlike lattice model simulations, structure prediction methods, and unfolding simulations, our simulations used an atom-based model, constant temperature (274 K), and non-beta-hairpin initial conformations. Based on the concept of solvent reference, the effective energy function simplified the solvent calculation and overcame the multiple minima problem. Driven by the hydrophobic interaction, the peptide first folded into a compact U shaped conformation with a central turn, in analogy to the initial collapse with simultaneous nucleation in protein folding. The peptide units in the U-shaped conformation then reoriented, gradually forming hydrogen bonds in the beta hairpin pattern from the beta-turn to the ends of the strands. With the same energy function, an alanine-based peptide folded into helix-dominated structures. The basic structure types (alpha-helix or beta-hairpin) that formed during the simulations depended upon the amino acid sequence. Compared with helix, beta hairpin folding is driven mainly by the hydrophobic interaction. Hydrogen bonding is necessary to maintain the ordered secondary structure. PMID- 9876132 TI - Adsorption of globular proteins on locally planar surfaces. II. Models for the effect of multiple adsorbate conformations on adsorption equilibria and kinetics. AB - Equilibrium and kinetic models for nonspecific adsorption of proteins to planar surfaces are presented. These models allow for the possibility of multiple interconvertible surface conformations of adsorbed protein. Steric repulsion resulting in area exclusion by adsorbed molecules is taken into account by treating the adsorbate as a thermodynamically nonideal two-dimensional fluid. In the equilibrium model, the possibility of attractive interactions between adsorbed molecules is taken into account in a limited fashion by permitting one of the adsorbed species to self-associate. Calculated equilibrium adsorption isotherms exhibit apparent high-affinity and low-affinity binding regions, corresponding respectively to adsorption of ligand at low fractional area occupancy in an energetically favorable side-on conformation and conversion at higher fractional area occupancy of the side-on conformation to an entropically favored end-on conformation. Adsorbate self-association may lead to considerable steepening of the adsorption isotherm, compensating to a variable extent for the broadening effect of steric repulsion. Kinetic calculations suggest that in the absence of attractive interactions between adsorbate molecules, the process of adsorption may be highly "stretched" along the time axis, rendering the attainment of adsorption equilibrium in the context of conventional experiments problematic. PMID- 9876133 TI - Unbinding of retinoic acid from its receptor studied by steered molecular dynamics. AB - Retinoic acid receptor (RAR) is a ligand-dependent transcription factor that regulates the expression of genes involved in cell growth, differentiation, and development. Binding of the retinoic acid hormone to RAR is accompanied by conformational changes in the protein which induce transactivation or transrepression of the target genes. In this paper we present a study of the hormone binding/unbinding process in order to clarify the role of some of the amino acid contacts and identify possible pathways of the all-trans retinoic acid binding/unbinding to/from human retinoic acid receptor (hRAR)-gamma. Three possible pathways were explored using steered molecular dynamics simulations. Unbinding was induced on a time scale of 1 ns by applying external forces to the hormone. The simulations suggest that the hormone may employ one pathway for binding and an alternative "back door" pathway for unbinding. PMID- 9876135 TI - A mutational analysis of the acetylcholine receptor channel transmitter binding site. AB - Mutagenesis and single-channel kinetic analysis were used to investigate the roles of four acetylcholine receptor channel (AChR) residues that are candidates for interacting directly with the agonist. The EC50 of the ACh dose-response curve was increased following alpha-subunit mutations Y93F and Y198F and epsilon subunit mutations D175N and E184Q. Single-channel kinetic modeling indicates that the increase was caused mainly by a reduced gating equilibrium constant (Theta) in alphaY198F and epsilonD175N, by an increase in the equilibrium dissociation constant for ACh (KD) and a reduction in Theta in alphaY93F, and only by a reduction in KD in epsilonE184Q. This mutation altered the affinity of only one of the two binding sites and was the only mutation that reduced competition by extracellular K+. Additional mutations of epsilonE184 showed that K+ competition was unaltered in epsilonE184D and was virtually eliminated in epsilonE184K, but that neither of these mutations altered the intrinsic affinity for ACh. Thus there is an apparent electrostatic interaction between the epsilonE184 side chain and K+ ( approximately 1.7kBT), but not ACh+. The results are discussed in terms of multisite and induced-fit models of ligand binding to the AChR. PMID- 9876136 TI - Frequency-dependent capacitance of the apical membrane of frog skin: dielectric relaxation processes. AB - Impedance analysis of the isolated epithelium of frog skin (northern Rana pipiens) was carried out in the frequency range between 0.1 Hz and 5.5 kHz while Na+ transport was abolished. Under these conditions, the impedance is determined almost completely by the dielectric properties of the apical membranes of the cells and the parallel shunt resistance. The modeling of the apical membrane impedance function required the inclusion of dielectric relaxation processes as originally described by. J. Chem. Phys. 9:341-351), where each process is characterized by a dielectric increment, relaxation frequency, and power law dependence. We found that the apical plasma membrane exhibited several populations of audio frequency dielectric relaxation processes centered at 30, 103, 2364, and 6604 Hz, with mean capacitive increments of 0.72, 1.00, 0.88, and 0.29 microF/cm2, respectively, that gave rise to dc capacitances of 1.95 +/- 0.06 microF/cm2 in 49 tissues. Capacitance was uncorrelated with large ranges of parallel shunt resistance and was not changed appreciably within minutes by K+ depolarization and hence a decrease in basolateral membrane resistance. A significant linear correlation existed between the dc capacitance and Na+ transport rates measured as short-circuit currents (Cadc = 0.028 Isc + 1.48; Isc between 4 and 35 microA/cm2) before inhibition of transport by amiloride and substitution of all Na+ with NMDG (N-methyl-D-glucamine) in the apical solution. The existence of dominant audio frequency capacitive relaxation processes complicates and precludes unequivocal interpretation of changes of capacitance in terms of membrane area alone when capacitance is measured at audio frequencies. PMID- 9876134 TI - Co-expression of lens fiber connexins modifies hemi-gap-junctional channel behavior. AB - Lens fiber cells contain two gap junction proteins (Cx56 and Cx45.6 in the chicken). Biochemical studies have suggested that these two proteins can form heteromeric connexons. To investigate the biophysical properties of heteromeric lens connexons, Cx56 was co-expressed with Cx45.6 (or its mouse counterpart, Cx50) in Xenopus oocytes. Whole-cell and single-channel currents were measured in single oocytes by conventional two-microelectrode voltage-clamp and patch clamp techniques, respectively. Injection of Cx56 cRNA induced a slowly activating, nonselective cation current that activated on depolarization to potentials higher than -10 mV. In contrast, little or no hemichannel current was induced by injection of Cx50 or Cx45.6 cRNA. Co-expression of Cx56 with Cx45.6 or Cx50 led to a shift in the threshold for activation to -40 or -70 mV, respectively. It also slowed the rate of deactivation of the hemichannel currents. Moreover, an increase in the unitary conductance, steady state probability of hemichannel opening and mean open times at negative potentials, was observed in (Cx56 + Cx45.6) cRNA-injected oocytes compared with Cx56 cRNA-injected oocytes. These results indicate that co-expression of lens fiber connexins gives rise to novel channels that may be explained by the formation of heteromeric hemichannels that contain both connexins. PMID- 9876137 TI - State-dependent cocaine block of sodium channel isoforms, chimeras, and channels coexpressed with the beta1 subunit. AB - Cocaine block of human cardiac (hH1) and rat skeletal (mu1) muscle sodium channels was examined under whole-cell voltage clamp in transiently transfected HEK293t cells. Low affinity block of resting mu1 and hH1 channels at -180 mV was the same, and high affinity block of inactivated channels at -70 mV was the same. Cocaine block of hH1 channels was greater than block of mu1 channels at voltages between -120 mV and -90 mV, suggesting that greater steady-state inactivation of hH1 channels in this voltage range makes them more susceptible to cocaine block. We induced shifts in the voltage dependence of steady-state inactivation at mu1 and hH1 channels by constructing mu1/hH1 channel chimeras or by coexpressing the wild-type channels with the rat brain beta1 subunit. In contrast to several previous reports, coexpression of the rat brain beta1 subunit with mu1 or hH1 produced large positive shifts in steady-state inactivation. Shifts in the voltage dependence of steady-state inactivation elicited linear shifts in steady state cocaine block, yet these manipulations did not affect the cocaine affinity of resting or inactivated channels. These data, as well as simulations used to predict block, indicate that state-dependent cocaine block depends on both steady state inactivation and channel activation, although inactivation appears to have the predominant role. PMID- 9876138 TI - Gbetagamma binding increases the open time of IKACh: kinetic evidence for multiple Gbetagamma binding sites. AB - IKACh is an inwardly rectifying potassium channel that plays an important role in the regulation of mammalian heart rate. IKACh is activated by direct interaction with Gbetagamma subunits of pertussis toxin-sensitive heterotrimeric G-proteins. The stoichiometry of the Gbetagamma/channel complex is currently unknown, and kinetic analysis of the channel behavior has led to conflicting conclusions. Here, we analyze the kinetics of the native IKACh channel in inside-out cardiomyocyte patches activated directly by Gbetagamma. We conclude that the channel has at least two open states and that binding of Gbetagamma prolongs its mean open time duration. These findings imply the existence of at least two binding sites on the channel complex for Gbetagamma. We also show that the duration of the channel opening is negatively correlated with the duration of subsequent channel closing, which further constrains the possible kinetic models. A simple qualitative model describing the kinetic behavior of IKACh is presented. PMID- 9876139 TI - Contribution of the selectivity filter to inactivation in potassium channels. AB - Voltage-gated K+ channels exhibit a slow inactivation process, which becomes an important influence on the rate of action potential repolarization during prolonged or repetitive depolarization. During slow inactivation, the outer mouth of the permeation pathway undergoes a conformational change. We report here that during the slow inactivation process, the channel progresses through at least three permeation states; from the initial open state that is highly selective for K+, the channel enters a state that is less permeable to K+ and more permeable to Na+, and then proceeds to a state that is non-conducting. Similar results were obtained in three different voltage-gated K+ channels: Kv2.1, a channel derived from Shaker (Shaker Delta A463C), and a chimeric channel derived from Kv2.1 and Kv1.3 that displays classical C-type inactivation. The change in selectivity displayed both voltage- and time-dependent properties of slow inactivation and was observed with K+ on either side of the channel. Elevation of internal [K+] inhibited Na+ conduction through the inactivating channel in a concentration dependent manner. These results indicate that the change in selectivity filter function is an integral part of the slow inactivation mechanism, and argue against the hypothesis that the inactivation gate is independent from the selectivity filter. Thus, these data suggest that the selectivity filter is itself the inactivation gate. PMID- 9876140 TI - Electron-spin resonance study of aggregation of gramicidin in dipalmitoylphosphatidylcholine bilayers and hydrophobic mismatch. AB - The effect of aggregation of gramicidin A' (GA) on the phase structure of dipalmitoylphosphatidylcholine (DPPC) multilamellar vesicles was studied by cw ESR using a chain-labeled lipid (16PC) at temperatures between 30 degrees and 45 degreesC that span the main phase transition of DPPC. Boundary lipids were observed only in dispersions with GA/DPPC molar ratios >1:15, where GA aggregates. Detailed fits by nonlinear least squares (NLLS) methods are consistent with the boundary lipid being characterized by a large negative order parameter ( approximately -0.4), indicative of a dynamic bending of the end of the acyl chain, and a substantially reduced motion, about an order of magnitude slower than that of the bulk lipid. The NLLS analysis compares favorably with a recent two-dimensional Fourier transform ESR study on DPPC/GA vesicles, which accurately discerned the bulk lipid. The detailed ESR observables are discussed in terms of the ordering effect of GA at low concentration of GA, the dissociation of the GA channel and the dynamic bending of the end chain segment of boundary lipid at high concentration of GA, and of HII phase formation induced by GA. It is suggested that these phenomena can be interpreted in terms of the combined effects of partial dehydration of the lipid headgroup by the GA and of the hydrophobic mismatch between GA and DPPC molecules. Substantial hysteresis is observed for heating versus cooling cycles, but only for a GA/DPPC molar ratio >1:15. This is consistent with the aggregation of GA molecules at high concentrations. PMID- 9876141 TI - The membrane-permeabilizing effect of avenacin A-1 involves the reorganization of bilayer cholesterol. AB - Avenacin A-1 is a member of a group of naturally occurring compounds called saponins. It is found in oat plants, where it protects against fungal pathogens. A combined electrical and optical chamber was used to determine the interaction of avenacin A-1 with Montal-Mueller planar lipid bilayers. This system allowed simultaneous measurement of the effect of avenacin A-1 on the fluorescence and lateral diffusion of a fluorescent lipid probe and permeability of the planar lipid bilayer. As expected, cholesterol was required for avenacin A-1-induced bilayer permeabilization. The planar lipid bilayers were also challenged with monodeglucosyl, bis-deglucosyl, and aglycone derivatives of avenacin A-1. The results show that the permeabilizing activity of the native avenacin A-1 was completely abolished after one, two, or all three sugar residues are hydrolyzed (monodeglucosyl, bis-deglucosyl, and aglycone derivatives, respectively). Fluorescence recovery after photobleaching (FRAP) measurements on cholesterol containing planar lipid bilayers revealed that avenacin A-1 caused a small but significant reduction in the lateral diffusion of the phospholipid probe N-(7 nitrobenzoyl-2-oxa-1,3-diazol-4-yl)-1, 2-dihexadecanoyl-sn-glycero-3 phosphoethanolamine (NBD-PE). Similarly, with the sterol probe (22-(N-(7 nitrobenz-2-oxa-1, 3-diazol-4-yl)amino)-23,24-bisnor-5-cholen-3beta-ol (NBD Chol), avenacin A-1, but not its derivatives, caused a more pronounced reduction in the lateral diffusion than that observed with the phospholipid probe. The data indicate that an intact sugar moiety of avenacin A-1 is required to reorganize membrane cholesterol into pores. PMID- 9876143 TI - Weak dependence of mobility of membrane protein aggregates on aggregate size supports a viscous model of retardation of diffusion. AB - Proteins in plasma membranes diffuse more slowly than proteins inserted into artificial lipid bilayers. On a long-range scale (>250 nm), submembrane barriers, or skeleton fences that hinder long-range diffusion and create confinement zones, have been described. Even within such confinement zones, however, diffusion of proteins is much slower than predicted by the viscosity of the lipid. The cause of this slowing of diffusion on the micro scale has not been determined and is the focus of this paper. One way to approach this question is to determine the dependence of particle motion on particle size. Some current models predict that the diffusion coefficient of a membrane protein aggregate will depend strongly on its size, while others do not. We have measured the diffusion coefficients of membrane glycoprotein aggregates linked together by concanavalin A molecules bound to beads of various sizes, and also the diffusion coefficients of individual concanavalin A binding proteins. The measurements demonstrate at most a weak dependence of diffusion coefficient on aggregate size. This finding supports retardation by viscous effects, and is not consistent with models involving direct interaction of diffusing proteins with cytoskeletal elements. PMID- 9876144 TI - Order in phospholipid Langmuir-Blodgett layers and the effect of the electrical potential of the substrate. AB - The ordering in dipalmitoylphosphatidylcholine (DPPC) Langmuir-Blodgett monolayers and bilayers on a semiconducting indium tin oxide (ITO) surface has been investigated at the equilibrium potential of the interface and at various externally applied potentials. Second- and fourth-rank order parameters of a diphenylhexatriene (DPH) containing phospholipid probe were derived from total internal reflection fluorescence measurements, and orientation distributions were calculated using the maximum-entropy method. Generally, bimodal orientation distributions were obtained, suggesting that only part of the probes is aligned with the DPPC molecules. The effect of applied potentials is small for DPPC layers on unmodified (hydrophilic) ITO; with decreasing potential the ordering changes slightly to more random distributions, possibly because of the onset of hydrogen evolution at the substrate surface. For monolayers on hydrophobized ITO, where the phospholipids are initially with their tails directed toward the surface, the changes are more significant. At the highest positive potential applied, the derived order parameters indicate that nearly all probes are flat on the surface. This can be understood as a result of enhanced competition between headgroups and tails for access to the surface as it becomes more polarized. On unmodified ITO the electrochemistry of Fe(CN)6(3-/4-) and Ru(bipyridyl)3(2+/3+) is hardly hindered by the presence of DPPC monolayers or bilayers. On hydrophobized ITO a DPPC monolayer enhances the redox reactions. PMID- 9876142 TI - The modified stalk mechanism of lamellar/inverted phase transitions and its implications for membrane fusion. AB - A model of the energetics of lipid assemblies (Siegel. 1993. Biophys. J. 65:2124 2140) is used to predict the relative free energy of intermediates in the transitions between lamellar (Lalpha) inverted hexagonal (HII), and inverted cubic (QII) phases. The model was previously used to generate the modified stalk theory of membrane fusion. The modified stalk theory proposes that the lowest energy structures to form between apposed membranes are the stalk and the transmonolayer contact (TMC), respectively. The first steps in the Lalpha/HII and Lalpha/QII phase transitions are also intermembrane events: bilayers of the Lalpha phase must interact to form new topologies during these transitions. Hence the intermediates in these phase transitions should be similar to the intermediates in the modified stalk mechanism of fusion. The calculations here show that stalks and TMCs can mediate transitions between the Lalpha, QII, and HII phases. These predictions are supported by studies of the mechanism of these transitions via time-resolved cryoelectron microscopy (. Biophys. J. 66:402-414; Siegel and Epand. 1997. Biophys. J. 73:3089-3111), whereas the predictions of previously proposed transition mechanisms are not. The model also predicts that QII phases should be thermodynamically stable in all thermotropic lipid systems. The profound hysteresis in Lalpha/QII transitions in some phospholipid systems may be due to lipid composition-dependent effects other than differences in lipid spontaneous curvature. The relevant composition-dependent properties are the Gaussian curvature modulus and the membrane rupture tension, which could change the stability of TMCs. TMC stability also influences the rate of membrane fusion of apposed bilayers, so these two properties may also affect the fusion rate in model membrane and biomembrane systems. One way proteins catalyze membrane fusion may be by making local changes in these lipid properties. Finally, although the model identifies stalks and TMCs as the lowest energy intermembrane intermediates in fusion and lamellar/inverted phase transitions, the stalk and TMC energies calculated by the present model are still large. This suggests that there are deficiencies in the current model for intermediates or intermediate energies. The possible nature of these deficiencies is discussed. PMID- 9876145 TI - Calcium-dependent binding of rabbit C-reactive protein to supported lipid monolayers containing exposed phosphorylcholine group. AB - The interaction of rabbit C-reactive protein (rCRP) with a supported monolayer containing a phosphorylcholine moiety was studied. Three types of phospholipids were synthesized, each containing a insertion spacer of eight, six, or three atoms between the phosphorylcholine group and hydrophobic tail. By varying the length of the insertion spacer, we can vary the extension of the phosphorylcholine group from the membrane surface. By varying the monolayer composition, we can control the lateral distance between the exposed phosphorylcholine groups. Using the surface plasmon resonance technique (SPR), we demonstrated that the calcium-dependent binding of rCRP to the model membrane is governed not only by the ability of the ligand to access the binding pocket fully (spacer length), but also by lateral hindrance within the two-dimensional plane of the membrane. The value of the apparent binding constant was estimated by theoretical analysis, which is obviously dependent on the composition of the lipid mixture, and a maximum of (9.9 +/- 1.5) x 10(6) M-1 was obtained. PMID- 9876146 TI - Ceramides in phospholipid membranes: effects on bilayer stability and transition to nonlamellar phases. AB - The effects of ceramides of natural origin on the gel-fluid and lamellar-inverted hexagonal phase transitions of phospholipids (mainly dielaidoylphosphatidylethanolamine) have been studied by differential scanning calorimetry, with additional support from infrared and 31P nuclear magnetic resonance (NMR) spectroscopy. In the lamellar phase, ceramides do not mix ideally with phospholipids, giving rise to the coexistence of domains that undergo the gel-fluid transition at different temperatures. The combination of differential scanning calorimetry and infrared spectroscopy, together with the use of deuterated lipids, allows the demonstration of independent melting temperatures for phospholipid and ceramide in the mixtures. In the lamellar-hexagonal phase transitions, ceramides (up to 15 mol %) decrease the transition temperature, without significantly modifying the transition enthalpy, thus facilitating the inverted hexagonal phase formation. 31P-NMR indicates the coexistence, within a certain range of temperatures, of lamellar and hexagonal phases, or hexagonal phase precursors. Ceramides from egg or from bovine brain are very similar in their effects on the lamellar-hexagonal transition. They are also comparable to diacylglycerides in this respect, although ceramides are less potent. These results are relevant in the interpretation of certain forms of interfacial enzyme activation and in the regulation and dynamics of the bilayer structure of cell membranes. PMID- 9876147 TI - Cholesterol orientation and dynamics in dimyristoylphosphatidylcholine bilayers: a solid state deuterium NMR analysis. AB - Proton decoupled deuterium NMR spectra of oriented bilayers made of DMPC and 30 mol % deuterated cholesterol acquired at 76.8 MHz (30 degreesC) have provided a set of very accurate quadrupolar splitting for eight C-D bonds of cholesterol. Due to the new precision of the experimental data, the original analysis by. Biochemistry. 23:6062-6071) had to be reconsidered. We performed a systematic study of the influence on the precision and uniqueness of the data-fitting procedure of: (i) the coordinates derived from x-ray, neutron scattering, or force field-minimized structures, (ii) internal mobility, (iii) the axial symmetry hypothesis, and (iv) the knowledge of some quadrupolar splitting assignments. Good agreement between experiment and theory could be obtained only with the neutron scattering structure, for which both axial symmetry hypothesis and full order parameter matrix analysis gave satisfactory results. Finally, this work revealed an average orientation of cholesterol slightly different from those previously published and, most importantly, a molecular order parameter equal to 0.95 +/- 0.01, instead of 0.79 +/- 0.03 previously found for the same system at 30 degreesC. Temperature dependence in the 20-50 degreesC range shows a constant average orientation and a monotonous decrease of cholesterol Smol, with a slope of -0.0016 K-1. A molecular order parameter of 0.89 +/- 0.01 at 30 degreesC was determined for a DMPC/16 mol % of cholesterol. PMID- 9876148 TI - Electrophysiological study with oxonol VI of passive NO3- transport by isolated plant root plasma membrane. AB - In contrast to animal cells, plant cells contain approximately 5-50 mM nitrate in cytosol and vacuole. The lack of specific spectroscopic probes, or suitable isotopes, impedes in vitro studies of NO3- transport. Reconstitution of root cell plasma membrane (PM) proteins in mixed soybean lipid:egg phosphatidylcholine allowed for the generation of large K+-valinomycin diffusion potentials (Em), monitored with the oxonol VI dye. Nevertheless, Em was restricted to approximately 130 mV by capacitor properties of biological membranes. This caused an increasing discrepancy at higher K+-Nernst potentials used for calibration. Therefore, Em was determined directly from the fluorescence of the dye free in buffer, bound at zero Em, and bound upon Em generation. Then, an electrophysiological analysis of the NO3--dependent dissipation rate of Em gave the net passive flux (JN) and the permeability coefficient to NO3- (PN). The plant root cell PM exhibited a strikingly large PN (higher than 10(-9) m s-1) at high Em (90-100 mV) and pH 6.5. At low Em (50-60 mV) and pH 7.4, PN decreased by 70-fold and became similar to that of the lipid bilayer. This agreed with the previous observation that 15 mM NO3- short-circuits the plant root PM H+-ATPase at its optimal pH of 6.5. PMID- 9876149 TI - Membrane fusion in vesicles of oligomerizable lipids. AB - Membrane fusion has been examined in a model system of small unilamellar vesicles of synthetic lipids that can be oligomerized through the lipid headgroups. The oligomerization can be induced either in both bilayer leaflets or in the inner leaflet exclusively. Oligomerization leads to denser lipid headgroup packing, with concomitant reduction of lipid lateral diffusion and membrane permeability. As evidenced by lipid mixing assays, electron microscopy, and light scattering, calcium-induced fusion of the bilayer vesicles is strongly retarded and inhibited by oligomerization. Remarkably, oligomerization of only the inner leaflet of the bilayer is already sufficient to affect fusion. The efficiency of inhibition and retardation of fusion critically depend on the relative amount of oligomeric lipid present, on the concentration of calcium ions, and on temperature. Implications for the mechanism of bilayer membrane fusion are discussed in terms of lipid lateral diffusion and membrane curvature effects. PMID- 9876150 TI - Studies of phospholipid hydration by high-resolution magic-angle spinning nuclear magnetic resonance. AB - A sample preparation method using spherical glass ampoules has been used to achieve 1.5-Hz resolution in 1H magic-angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of aqueous multilamellar dispersions of 1,2-dioleoyl-sn glycero-3-phosphocholine (DOPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC), serving to differentiate between slowly exchanging interlamellar and bulk water and to reveal new molecular-level information about hydration phenomena in these model biological membranes. The average numbers of interlamellar water molecules in multilamellar vesicles (MLVs) of DOPC and POPC were found to be 37.5 +/- 1 and 37.2 +/- 1, respectively, at a spinning speed of 3 kHz. Even at speeds as high as 9 kHz, the number of interlamellar waters remained as high as 31, arguing against dehydration effects for DOPC and POPC. Both homonuclear and heteronuclear nuclear Overhauser enhancement spectroscopy (NOESY and HOESY) were used to establish the location of water near the headgroup of a PC bilayer. 1H NMR comparisons of DOPC with a lipid that can hydrogen bond (monomethyldioleoylphosphatidylethanolamine, MeDOPE) showed the following trends: 1) the interlamellar water resonance was shifted to lower frequency for DOPC but to higher frequency for MeDOPE, 2) the chemical shift variation with temperature for interlamellar water was less than that of bulk water for MeDOPE MLVs, 3) water exchange between the two lipids was rapid on the NMR time scale if they were mixed in the same bilayer, 4) water exchange was slow if they were present in separate MLVs, and 5) exchange between bulk and interlamellar water was found by two-dimensional exchange experiments to be slow, and the exchange rate should be less than 157 Hz. These results illustrate the utility of ultra-high resolution 1H MAS NMR for determining the nature and extent of lipid hydration as well as the arrangement of nuclei at the membrane/water interface. PMID- 9876152 TI - Estimating the persistence length of a worm-like chain molecule from force extension measurements. AB - We describe a simple computation of the worm-like chain model and obtain the corresponding force-versus-extension curve. We propose an improvement to the Marko and Siggia interpolation formula of Bustamante et al (Science 1994, 265:1599-1600) that is useful for fitting experimental data. We apply it to the experimental elasticity curve of single DNA molecules. Finally, we present a tool to study the agreement between the worm-like chain model and experiments. PMID- 9876153 TI - Heme-solvent coupling: a Mossbauer study of myoglobin in sucrose. AB - The Mossbauer effect of 57Fe-enriched samples was used to investigate the coupling of 80% sucrose/water, a protein-stabilizing solvent, to vibrational and diffusive modes of the heme iron of CO-myoglobin. For comparison we also determined the Mossbauer spectra of K4 57Fe (CN)6 (potassium ferrocyanide, PFC), where the iron is fully exposed in the same solvent. The temperature dependence of the Mossbauer parameters derived for the two samples proved to be remarkably similar, indicative of a strong coupling of the main heme displacements to the viscoelastic relaxation of the solvent. We show that CO escape out of the heme pocket couples to the same type of fluctuations, whereas intramolecular bond formation involves solvent-decoupled heme deformation modes that are less prominent in the Mossbauer spectrum. With respect to other solvents, however, sucrose shows a reduced viscosity effect on heme displacements and the kinetics of ligand binding due to preferential hydration of the protein. This result confirms thermodynamic predictions of the stabilizing action of sucrose by a dynamic method. PMID- 9876151 TI - Mutations in the N- and D-helices of the N-domain of troponin C affect the C domain and regulatory function. AB - Troponin C contains a 14-residue alpha-helix at the amino terminus, the N-helix, that calmodulin lacks. Deletion of the first 11-14 residues of troponin C alters function. In the present investigation a mutant lacking residues 1-7 of the N helix has normal conformation, Ca2+ binding, and regulatory function. Thus, residues 8-14 of the N-helix are generally sufficient for troponin C function. In the x-ray structures of troponin C there is a salt bridge between Arg 11 in the N helix and Glu 76 in the D-helix. Destroying the salt bridge by individually mutating the residues to Cys has no effect on function. However, mutation of both residues to Cys reduces troponin C's affinity for the troponin complex on the thin filament, reduces the stability of the N-domain in the absence of divalent cations, increases the Ca2+ affinity and reduces the cooperativity of the Ca2+Mg2+ sites in the C-domain, and alters the conformational change that takes place upon Ca2+ binding (but not Mg2+ binding) to the C-domain. Cross-linking with bis-(maleimidomethylether) partially restores function. The Ca2+-specific sites in the N-domain, those closest to the sites of the mutations, are unaffected in the assays employed. These results show that the N-helix is a critical structural element for interaction with and activation of the thin filament. Moreover, mutations in the N-helix affect the C-terminal domain, consistent with recent structural studies showing that the N-helix and C-terminal domain are physically close. PMID- 9876154 TI - Tetrameric assembly of full-sequence protein zero myelin glycoprotein by synchrotron x-ray scattering. AB - Highly purified myelin P0 glycoprotein was solubilized to 1-8 mg/ml in 0.1% sodium dodecyl sulfate (SDS), and the solution structure of the P0 assembly was studied using synchrotron x-ray scattering. The full-length P0, which was isolated from bovine intradural roots, included both the extracellular and cytoplasmic domains of the molecule. At the higher concentrations (4, 6, and 8 mg/ml, respectively), an x-ray intensity maximum was observed at 316 A, 245 A, and 240 A Bragg spacing. Because the position of this intensity depended on P0 concentration, it is most likely due to interparticle interference. By contrast, the position of a second intensity maximum, which was at approximately 40 A Bragg spacing, was invariant with P0 concentration. This latter intensity was accounted for by monodispersed, 80 A-diameter particles that are composed of eight, approximately 30 A-diameter spheres. Chemical parameters suggest that the 80 A particles correspond to the size of a tetramer of P0 molecules. Therefore, the approximately 30 A spheres would correspond to the sizes of the extracellular and cytoplasmic domains for each of the P0 monomers. The invariance of the second intensity maximum with P0 concentration indicates that the structure of the 80 A diameter, tetrameric particles is unaltered. According to the liquid model for interparticle interference from charged spheres, the 80 A-diameter particle has 10 negative surface charges which likely arise from negatively charged SDS molecules bound to the transmembrane domain of P0. This binding, however, apparently does not alter the tetrameric assembly of P0, suggesting that intermolecular interactions involving extracellular domains and cytoplasmic domains likely stabilize this assembly. Some of our results have been published in abstract form (Inouye, H., H. Tsuruta, D. A. Kirschner, J. Sedzik, and K. Uyemura. Abstracts of the 4th International School and Symposium on Synchrotron Radiation in Natural Science, June 15-20, 1998. Ustron-Jaszowiec, Poland. p. 31). PMID- 9876155 TI - Electron transfer kinetics of caa3 oxidase from Bacillus stearothermophilus: a hypothesis for thermophilicity. AB - The O2 reaction and the reverse electron transfer of the thermophilic caa3 terminal oxidase of Bacillus stearothermophilus have been studied by laser flash photolysis. The results show that both reactions, although studied at a temperature of 20 degreesC, far from the optimal temperature of > 60 degreesC for caa3, follow a kinetic behavior essentially identical to that observed with the electrostatic complex between mammalian cyt c and cyt c oxidase. In the O2 reaction cyt a and cyt a3 are very quickly oxidized; cyt a is then re-reduced via CuA, whereas cyt c oxidation is apparently rate-limited by the oxidation of CuA. Upon photodissociation of the mixed valence-CO caa3, reverse electron transfer from the binuclear center to cyt a3+ (tau1 = 3 micros) and CuA2+ (tau2 = 64 micros) is observed, while cyt c is not reduced by any detectable level. These results seem to rule out accounting for enzymatic thermophilicity by altered kinetics of intramolecular electron transfer involving the cyt center in the reduced configuration, which is very fast. On the basis of these results and previous data, we propose that thermophilicity involves an increased activation barrier for the reduction of cyt a3-CuB in the configuration typical of the oxidized site. PMID- 9876156 TI - Apohorseradish peroxidase unfolding and refolding: intrinsic tryptophan fluorescence studies. AB - The unfolding and refolding of apohorseradish peroxidase, as a function of guanidinium chloride concentration, were monitored by the intrinsic fluorescence intensity, polarization, and lifetime of the single tryptophan residue. The unfolding was reversible and characterized by at least three distinct stages-the intensity and lifetime data, for example, were both characterized by an initial increase followed by a decrease and then a plateau region. The lifetime data, in the absence and presence of guanidinium chloride, were heterogeneous and fit best to a model consisting of a major Gaussian distribution component and a minor, short discrete component. The observed increase in intensity in the initial stage of the unfolding process is attributed to the conversion of this short component into the longer, distributed component as the guanidinium chloride concentration increases. Our results clarify and amplify previous studies on the unfolding of apohorseradish peroxidase by guanidinium chloride. PMID- 9876157 TI - Two-phase induction of the nonnative alpha-helical form of beta-lactoglobulin in the presence of trifluoroethanol. AB - The trifluoroethanol-dependent induction of the nonnative alpha-helical form of beta-lactoglubulin has been studied by circular dichroism spectroscopy. Data analysis is performed by factor analysis and multivariate curve resolution. An intermediate form in the induction of the alpha-helical form of the beta lactoglobulin has been identified at low TFE concentration. By application of an alternating least-squares algorithm, the CD spectrum corresponding to the intermediate form has been resolved. The deconvolution of this CD spectrum shows a secondary structure content more in agreement with the one predicted from the amino acid sequence than the secondary structure of the helical form obtained at higher TFE concentrations. The additional alpha-helical content of the form present at higher TFE concentrations could be due to nonspecific interaction of TFE with the polypeptide chain. PMID- 9876158 TI - Bacterial S-layer protein coupling to lipids: x-ray reflectivity and grazing incidence diffraction studies. AB - The coupling of bacterial surface (S)-layer proteins to lipid membranes is studied in molecular detail for proteins from Bacillus sphaericus CCM2177 and B. coagulans E38-66 recrystallized at dipalmitoylphosphatidylethanolamine (DPPE) monolayers on aqueous buffer. A comparison of the monolayer structure before and after protein recrystallization shows minimal reorganization of the lipid chains. By contrast, the lipid headgroups show major rearrangements. For the B. sphaericus CCM2177 protein underneath DPPE monolayers, x-ray reflectivity data suggest that amino acid side chains intercalate the lipid headgroups at least to the phosphate moieties, and probably further beyond. The number of electrons in the headgroup region increases by more than four per lipid. Analysis of the changes of the deduced electron density profiles in terms of a molecular interpretation shows that the phosphatidylethanolamine headgroups must reorient toward the surface normal to accommodate such changes. In terms of the protein structure (which is as yet unknown in three dimensions), the electron density profile reveals a thickness lz approximately 90 A of the recrystallized S-layer and shows water-filled cavities near its center. The protein volume fraction reaches maxima of >60% in two horizontal sections of the S-layer, close to the lipid monolayer and close to the free subphase. In between it drops to approximately 20%. Four S-layer protein monomers are located within the unit cell of a square lattice with a spacing of approximately 131 A. PMID- 9876159 TI - Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives. AB - We investigated the use of rhodamine 123 (R123), tetramethylrhodamine methyl ester (TMRM), and tetramethylrhodamine ethyl ester (TMRE) as fluorescent probes to monitor the membrane potential of mitochondria. These indicator dyes are lipophilic cations accumulated by mitochondria in proportion to DeltaPsi. Upon accumulation, all three dyes exhibit a red shift in both their absorption and fluorescence emission spectra. The fluorescence intensity is quenched when the dyes are accumulated by mitochondria. These properties have been used to develop a method to dynamically monitor DeltaPsi of isolated rat heart mitochondria using a ratio fluorescence approach. All three dyes bound to the inner and outer aspects of the inner mitochondrial membrane and, as a result, were accumulated by mitochondria in a greater quantity than predicted by the Nernst equation. Binding to mitochondria was temperature-dependent and the degree of binding was in the order of TMRE > R123 > TMRM. The internal and external partition coefficients for binding were determined to correct for binding in the calculation of DeltaPsi. All three dyes suppressed mitochondrial respiratory control to some extent. Inhibition of respiration was greatest with TMRE, followed by R123 and TMRM. When used at low concentrations, TMRM did not suppress respiration. The use of these dyes and ratio fluorescence techniques affords a simple method for measurement of DeltaPsi of isolated mitochondria. We also applied this approach to the isolated perfused heart to determine whether DeltaPsi could be monitored in an intact tissue. Wavelength scanning of the surface fluorescence of the heart under various conditions after accumulation of TMRM indicated that the mitochondrial matrix-induced wavelength shift of TMRM also occurs in the heart cytosol, eliminating the use of this approach in the intact heart. PMID- 9876160 TI - Does pressure antagonize anesthesia? High-pressure stopped-flow study of firefly luciferase and anatomy of initial flash. AB - The antagonizing effect of high pressure against anesthesia is well known. With purified firefly luciferase, however,. Biophys. J. 60:1309-1314) reported that high pressure did not affect the initial flash intensity. Firefly luciferase emits a burst of light when the substrates luciferin and ATP are added in the presence of O2. The light intensity decays rapidly and the weak light lasts for hours. The initial flash is a transient event and is not in a steady state. The steady state is represented by the slope of the linear part of the integral of the light output. The present study used a high-pressure stopped-flow system to compare the pressure effects on the initial flash intensity and the steady-state light intensity. The flash intensity did not change by the application of hydrostatic pressure in the presence or absence of chloroform or 1-octanol. In contrast, high pressure increased the steady-state light intensity. The application of 12 MPa pressure increased the steady-state light intensity of firefly luciferase inhibited by 5 mM chloroform or 0.7 mM 1-octanol by 19.7% and 18.8%, respectively. When analyzed by the rapid reaction kinetics of the transition state theory, the initial peak intensity represents the total amount of active enzyme and is unrelated to the reaction rate. Anesthetics inhibited the initial flash by unfolding the protein, thereby decreasing the concentration of the active enzyme. Pressure affected the steady-state light intensity by changing the reaction rates. PMID- 9876161 TI - Does pressure antagonize anesthesia? Opposite effects on specific and nonspecific inhibitors of firefly luciferase. AB - Ueda and Suzuki (1998. Biochim. Biophys. Acta. 1380:313-319; 1998. Biophys. J. 75:1052-1057) reported that myristic acid inhibited firefly luciferase in microM range in competition with luciferin, whereas anesthetics inhibited it in millimeter ranges noncompetitively with luciferin. Myristate increased, whereas anesthetics decreased, the thermal denaturation temperature. The present study showed that high pressure increased the steady-state light intensity of the halothane-doped firefly luciferase but decreased that of the myristate-doped firefly luciferase. The steady-state light intensity showed a maximum at 19.1 degrees C. At 19.1 degrees C, high pressure did not affect the light intensity in the absence of the inhibitors. In the presence of 0.5 mM halothane, however, 25 MPa pressure (maximum effect) increased the light intensity to 106.0% of the control without the inhibitor. In the presence of 2.5 microM myristate, 40 MPa pressure decreased the light intensity to 90.9% of the control. When the temperature was 25 degrees C in the absence of inhibitors, 40 MPa pressure increased the light intensity 119.2% of the ambient value. At 0.5 mM halothane, 40 MPa pressure further increased the light intensity to 106.1% above the control 40 MPa value. At 2.5 microM myristate, 40 MPa pressure decreased the light intensity to 90.1% of the control 40 MPa value. From the pressure dependence of the light intensity, the volume change DeltaV of the enzyme was estimated at 25 degrees C: 0.5 mM halothane increased DeltaV = +3.93 cm3 mol-1, whereas 2.5 microM myristate decreased DeltaV = -7.66 cm3 mol-1. Present results show that there are distinct differences between the specific and nonspecific ligands in their response to high pressure. Myristate, which competes with luciferin, decreased the protein volume and stabilized the conformation against thermal perturbation. Halothane, which does not compete with the substrate, increased the protein volume and destabilized the conformation. PMID- 9876162 TI - Photolysis of caged calcium in femtoliter volumes using two-photon excitation. AB - A new technique for the determination of the two-photon uncaging action cross section (deltau) of photolyzable calcium cages is described. This technique is potentially applicable to other caged species that can be chelated by a fluorescent indicator dye, as well as caged fluorescent compounds. The two-photon action cross sections of three calcium cages, DM-nitrophen, NP-EGTA, and azid-1, are studied in the range of excitation wavelengths between 700 and 800 nm. Azid-1 has a maximum deltau of approximately 1.4 GM at 700 nm, DM-nitrophen has a maximum deltau of approximately 0.013 GM at 730 nm, and NP-EGTA has no measurable uncaging yield. The equations necessary to predict the amount of cage photolyzed and the temporal behavior of the liberated calcium distribution under a variety of conditions are derived. These equations predict that by using 700-nm light from a Ti:sapphire laser focused with a 1.3-NA objective, essentially all of the azid-1 within the two-photon focal volume would be photolyzed with a 10-micros pulse train of approximately 7 mW average power. The initially localized distributions of free calcium will dissipate rapidly because of diffusion of free calcium and uptake by buffers. In buffer-free cytoplasm, the elevation of the calcium concentration at the center of the focal volume is expected to last for approximately 165 micros. PMID- 9876163 TI - Reflection interference contrast microscopy combined with scanning force microscopy verifies the nature of protein-ligand interaction force measurements. AB - The integration of a stand-alone scanning force microscope (SFM) scanner with a reflection interference contrast microscope (RICM) makes it possible to measure directly the separation distance between the SFM probe and the sample surface. The SFM-RICM combination, when applied to the force measurements between ligand derivatized SFM probe and a protein receptor-derivatized surface, showed that the anomalous force discontinuities often observed for such interacting pairs were indeed a real behavior characteristic of a particular experimental configuration. Apart from small discrepancies due to transient damping, commercially available cantilevers did behave in an ideal mechanical fashion, thus indicating that protein-ligand unbinding events were occurring at distances much larger than their maximum extended length. This external verification of separation distance requires a closer examination of the physical events occurring upon detachment of the surfaces. An alternative interpretation of such force measurements is proposed here in which the protein and/or ligand immobilization chemistry is called into question. PMID- 9876164 TI - Imaging of cell/substrate contacts of living cells with surface plasmon resonance microscopy. AB - We have developed a new method for observing cell/substrate contacts of living cells in culture based on the optical excitation of surface plasmons. Surface plasmons are quanta of an electromagnetic wave that travel along the interface between a metal and a dielectric layer. The evanescent field associated with this excitation decays exponentially perpendicular to the interface, on the order of some hundreds of nanometers. Cells were cultured on an aluminum-coated glass prism and illuminated from below with a laser beam. Because the cells interfere with the evanescent field, the intensity of the reflected light, which is projected onto a camera chip, correlates with the cell/substrate distance. Contacts between the cell membrane and the substrate can thus be visualized at high contrast with a vertical resolution in the nanometer range. The lateral resolution along the propagation direction of surface plasmons is given by their lateral momentum, whereas perpendicular to it, the resolution is determined by the optical diffraction limit. For quantitative analysis of cell/substrate distances, cells were imaged at various angles of incidence to obtain locally resolved resonance curves. By comparing our experimental data with theoretical surface plasmon curves we obtained a cell/substrate distance of 160 +/- 10 nm for most parts of the cells. Peripheral lamellipodia, in contrast, formed contacts with a cell substrate/distance of 25 +/- 10 nm. PMID- 9876165 TI - Analysis of vertical fluorescence resonance energy transfer from the surface of a small-diameter sphere. AB - Fluorescence resonance energy transfer (FRET) measurements have been used to analyze fluorophore separations in a number of varying geometries, including small particles and extended surfaces. This study focuses on the geometry created by a donor extended above the surface of a small sphere (radius < R0), where the acceptors are integrated into the sphere surface. The model of this geometry was based on an amphipathic molecule with its lipophilic region integrated into a detergent micelle and its hydrophilic region extending outward from the micelle surface, where the donor fluorophore is attached to the hydrophilic region of the molecule. Based on random acceptor incorporation into the micelle, a Poisson distribution was used to calculate the distribution of acceptor probes across the micelle population. The model converges to RET on a flat surface when the radius of the micelle exceeds 0.8 R0. The model was also used to simulate FRET data showing that the positions of donors above the micelle surface could be uniquely resolved. Experimental verification of the model was achieved in a sulfobetaine palmitate micelle with fluorescein isothiocyanate donors attached to detergent solubilized lipopolysaccharide (LPS) and lipophilic Fast-DiI acceptors. The use of steady-state analysis allowed resolution of cases in which donors were located at different distances from the surface. Combining steady-state with excited state lifetime analysis allowed resolution of cases where there was a combination of distances. Given the large number of biomolecules that interact with lipids, this approach may prove generally useful for defining molecular conformation. PMID- 9876166 TI - Relative surface charge density mapping with the atomic force microscope. AB - An experimental approach for producing relative charge density maps of biological surfaces using the atomic force microscope is presented. This approach, called D minus D (D-D) mapping, uses isoforce surfaces collected at different salt concentrations to remove topography and isolate electrostatic contributions to the tip-sample interaction force. This approach is quantitative for surface potentials below 25 mV, and does not require prior knowledge of the cantilever spring constant, tip radius, or tip charge. In addition, D-D mapping does not require tip-sample contact. The performance of D-D mapping is demonstrated on surfaces of constant charge and varying topography (mechanically roughened mica and stacked bilayers of dipalmitolphosphatidylserine), a surface of varying charge and varying topography (patches of dipalmitolphosphatidylcholine on mica), and bacteriorhopsin membranes adsorbed to mica. PMID- 9876167 TI - Quantitative orientation measurements in thin lipid films by attenuated total reflection infrared spectroscopy. AB - Quantitative orientation measurements by attenuated total reflectance (ATR) infrared spectroscopy require the accurate knowledge of the dichroic ratio and of the mean-square electric fields along the three axes of the ATR crystal. In this paper, polarized ATR spectra of single supported bilayers of the phospholipid dimyristoylphosphatidic acid covered by either air or water have been recorded and the dichroic ratio of the bands due to the methylene stretching vibrations has been calculated. The mean-square electric field amplitudes were calculated using three formalisms, namely the Harrick thin film approximation, the two-phase approximation, and the thickness- and absorption-dependent one. The results show that for dry bilayers, the acyl chain tilt angle varies with the formalism used, while no significant variations are observed for the hydrated bilayers. To test the validity of the different formalisms, s- and p-polarized ATR spectra of a 40 A lipid layer were simulated for different acyl chain tilt angles. The results show that the thickness- and absorption-dependent formalism using the mean values of the electric fields over the film thickness gives the most accurate values of acyl chain tilt angle in dry lipid films. However, for lipid monolayers or bilayers, the tilt angle can be determined with an acceptable accuracy using the Harrick thin film approximation. Finally, this study shows clearly that the uncertainty on the determination of the tilt angle comes mostly from the experimental error on the dichroic ratio and from the knowledge of the refractive index. PMID- 9876168 TI - Membrane helix orientation from linear dichroism of infrared attenuated total reflection spectra. AB - Oriented multilamellar systems containing phospholipids and peptides have been formed on a germanium internal reflection element. Attenuated total reflection infrared spectra have been recorded and the linear dichroism of peptide amide I and amide II bands measured. Using peptides for which the orientation had been previously studied under similar experimental conditions by 15N solid-state nuclear magnetic resonance spectroscopy, important conclusions were drawn on the approach to be used to derive secondary structure orientation in a membrane from dichroic ratios. In particular, it is shown that the influence of the film thickness and refractive index on the orientation determination can be evaluated from the value of RATRiso, i.e., the dichroic ratio of a dipole oriented at the magic angle or with isotropic mobility. A series of peptides was used to test the validity of our suggestions on various helix orientations in the membrane. These include magainin 2 and hydrophobic (hPhi20) model peptides, the transmembrane segment of glycophorin (GLY), and LAH4, a designed peptide antibiotic that changes between a transmembrane and an in-plane orientation in a pH-dependent manner. PMID- 9876169 TI - Inclusions on fluid membranes anchored to elastic media. AB - We model theoretically the effect of localized forces on a fluid membrane anchored to a uniform elastic medium. We use this as a simple model for the plasma membrane of a cell. The atomic force microscope (AFM) has been used to apply such forces, but large membrane perturbations occurring in vivo are also treated within the same framework. Inclusions of this nature may include cell junctions, filipodia, caveolae, and similar membrane invaginations. The breakdown of linear elastic response, as observed by AFM, is predicted to occur for forces as small as 10 pN. We estimate the position of this crossover and the subsequent nonlinear behavior and make encouraging quantitative comparison with experiments. Intrinsic membrane inclusions interact through their overlapping strain fields. For similar, point force-like inclusions at large separations, this yields an attractive potential that scales like the inverse of their separation. For membranes that are intrinsically stiff or under tension, the binding force between inclusions can depend on the properties of the membrane and may be large enough to induce aggregation of inclusions, as observed experimentally. For inclusions that fix the magnitude of the membrane deformation, rather than the applied force, we demonstrate the possibility of metastable states, corresponding to finite separations. Finally, we discuss briefly the case in which inclusions couple to the membrane in more complex ways, such as via a torque (twist). In such cases, the interaction scales like a higher power of the separation, depends on the orientation of the inclusions, and can have either sign. PMID- 9876170 TI - Measurement of local viscoelasticity and forces in living cells by magnetic tweezers. AB - We measured the viscoelastic properties of the cytoplasm of J774 macrophages with a recently developed microrheometer. Ferromagnetic beads (1.3 microm in diameter) were used to determine the local viscoelastic moduli. Step-force pulses were applied to the magnetic beads and the displacement was observed by single particle tracking. By analyzing the creep response curves in terms of a triphasic mechanical equivalent circuit, we measured the shear elastic modulus, the effective viscosities, and the strain relaxation time. The values of the shear modulus vary by more than an order of magnitude within the cell population (range, 20-735 Pa; average, 343 Pa) and by a factor of 2 within single cells. The effective viscosity of the cytoplasm exhibits a relatively sharp distribution about an average of eta = 210 Pa s (+/- 143 Pa s). We measured the displacement field generated by the local forces by observing the induced motion of nonmagnetic beads. Even at distances of the order of 1 microm, no induced motion was seen, suggesting that the cytoplasm is composed of clusters of densely packed and cross-linked filaments separated by soft regions. In another series of experiments we analyzed the magnetophoretic motion of the ferromagnetic beads at a constant magnetic force. Measuring the bead velocity parallel and perpendicular to the applied force showed that local active forces on the beads varied from 50 to 900 pN. PMID- 9876171 TI - Torque generated by the flagellar motor of Escherichia coli while driven backward. AB - The technique of electrorotation was used to apply torque to cells of the bacterium Escherichia coli tethered to glass coverslips by single flagella. Cells were made to rotate backward, that is, in the direction opposite to the rotation driven by the flagellar motor itself. The torque generated by the motor under these conditions was estimated using an analysis that explicitly considers the angular dependence of both the viscous drag coefficient of the cell and the torque produced by electrorotation. Motor torque varied approximately linearly with speed up to over 100 Hz in either direction, placing constraints on mechanisms for torque generation in which rates of proton transfer for backward rotation are limiting. These results, interpreted in the context of a simple three-state kinetic model, suggest that the rate-limiting step in the torque generating cycle is a powerstroke in which motor rotation and dissipation of the energy available from proton transit occur synchronously. PMID- 9876172 TI - The origins of primitive blood in Xenopus: implications for axial patterning. AB - The marginal zone in Xenopus laevis is proposed to be patterned with dorsal mesoderm situated near the upper blastoporal lip and ventral mesoderm near the lower blastoporal lip. We determined the origins of the ventralmost mesoderm, primitive blood, and show it arises from all vegetal blastomeres at the 32-cell stage, including blastomere C1, a progenitor of Spemann's organizer. This demonstrates that cells located at the upper blastoporal lip become ventral mesoderm, not solely dorsal mesoderm as previously believed. Reassessment of extant fate maps shows dorsal mesoderm and dorsal endoderm descend from the animal region of the marginal zone, whereas ventral mesoderm descends from the vegetal region of the marginal zone, and ventral endoderm descends from cells located vegetal of the bottle cells. Thus, the orientation of the dorsal-ventral axis of the mesoderm and endoderm is rotated 90( degrees) from its current portrayal in fate maps. This reassessment leads us to propose revisions in the nomenclature of the marginal zone and the orientation of the axes in pre-gastrula Xenopus embryos. PMID- 9876173 TI - Cell death and cell protection genes determine the fate of pistils in maize. AB - The formation of unisexual flowers in maize requires the selective elimination and sexual maturation of floral organs in an initially bisexual floral meristem. Elimination of pistil primordia occurs in the primary and secondary florets of the tassel spikelets, and in the secondary florets of ear spikelets. Ill-fated pistil cells undergo a cell death process associated with nuclear degeneration in a specific spatial-temporal pattern that begins in the subepidermis, eventually aborting the entire organ. The sex determination genes tasselseed1 and tasselseed2 are required for death of pistil cells. tasselseed1 is required for the accumulation of TASSELSEED2 mRNA in pistil cells. All pistil primordia express TASSELSEED2 RNA but functional pistils found in ear spikelets are protected from cell death by the action of the silkless1 gene. silkless1 blocks tasselseed-induced cell death in the pistil primordia of primary ear florets. A model is proposed for the control of pistil fate by the action of the ts1-ts2-sk1 pathway. PMID- 9876174 TI - Transcription factors Mash-1 and Prox-1 delineate early steps in differentiation of neural stem cells in the developing central nervous system. AB - Like other tissues and organs in vertebrates, multipotential stem cells serve as the origin of diverse cell types during genesis of the mammalian central nervous system (CNS). During early development, stem cells self-renew and increase their total cell numbers without overt differentiation. At later stages, the cells withdraw from this self-renewal mode, and are fated to differentiate into neurons and glia in a spatially and temporally regulated manner. However, the molecular mechanisms underlying this important step in cell differentiation remain poorly understood. In this study, we present evidence that the expression and function of the neural-specific transcription factors Mash-1 and Prox-1 are involved in this process. In vivo, Mash-1- and Prox-1-expressing cells were defined as a transient proliferating population that was molecularly distinct from self renewing stem cells. By taking advantage of in vitro culture systems, we showed that induction of Mash-1 and Prox-1 coincided with an initial step of differentiation of stem cells. Furthermore, forced expression of Mash-1 led to the down-regulation of nestin, a marker for undifferentiated neuroepithelial cells, and up-regulation of Prox-1, suggesting that Mash-1 positively regulates cell differentiation. In support of these observations in vitro, we found specific defects in cellular differentiation and loss of expression of Prox-1 in the developing brain of Mash-1 mutant mice in vivo. Thus, we propose that induction of Mash-1 and Prox-1 is one of the critical molecular events that control early development of the CNS. PMID- 9876175 TI - Defective oligodendrocyte development and severe hypomyelination in PDGF-A knockout mice. AB - There is a class of oligodendrocyte progenitors, called O-2A progenitors, that is characterized by expression of platelet-derived growth factor &agr;-receptors (PDGFR(&agr;)). It is not known whether all oligodendrocytes are derived from these PDGFRalpha-progenitors or whether a subset(s) of oligodendrocytes develops from a different, PDGFR alpha-negative lineage(s). We investigated the relationship between PDGF and oligodendrogenesis by examining mice that lack either PDGF-A or PDGF-B. PDGF-A null mice had many fewer PDGFR alpha-progenitors than either wild-type or PDGF-B null mice, demonstrating that proliferation of these cells relies heavily (though not exclusively) on PDGF-AA homodimers. PDGF-A deficient mice also had reduced numbers of oligodendrocytes and a dysmyelinating phenotype (tremor). Not all parts of the central nervous system (CNS) were equally affected in the knockout. For example, there were profound reductions in the numbers of PDGFR alpha-progenitors and oligodendrocytes in the spinal cord and cerebellum, but less severe reductions of both cell types in the medulla. This correlation suggests a close link between PDGFRalpha-progenitors and oligodendrogenesis in most or all parts of the CNS. We also provide evidence that myelin proteolipid protein (PLP/DM-20)-positive cells in the late embryonic brainstem are non-dividing cells, presumably immature oligodendrocytes, and not proliferating precursors. PMID- 9876176 TI - The PINHEAD/ZWILLE gene acts pleiotropically in Arabidopsis development and has overlapping functions with the ARGONAUTE1 gene. AB - Several lines of evidence indicate that the adaxial leaf domain possesses a unique competence to form shoot apical meristems. Factors required for this competence are expected to cause a defect in shoot apical meristem formation when inactivated and to be expressed or active preferentially in the adaxial leaf domain. PINHEAD, a member of a family of proteins that includes the translation factor eIF2C, is required for reliable formation of primary and axillary shoot apical meristems. In addition to high-level expression in the vasculature, we find that low-level PINHEAD expression defines a novel domain of positional identity in the plant. This domain consists of adaxial leaf primordia and the meristem. These findings suggest that the PINHEAD gene product may be a component of a hypothetical meristem forming competence factor. We also describe defects in floral organ number and shape, as well as aberrant embryo and ovule development associated with pinhead mutants, thus elaborating on the role of PINHEAD in Arabidopsis development. In addition, we find that embryos doubly mutant for PINHEAD and ARGONAUTE1, a related, ubiquitously expressed family member, fail to progress to bilateral symmetry and do not accumulate the SHOOT MERISTEMLESS protein. Therefore PINHEAD and ARGONAUTE1 together act to allow wild-type growth and gene expression patterns during embryogenesis. PMID- 9876177 TI - Abrogation of the Cripto gene in mouse leads to failure of postgastrulation morphogenesis and lack of differentiation of cardiomyocytes. AB - Cripto-1(Cr1) protein encoded by the tdgf1 gene, is a secreted growth factor that is expressed early in embryonic development and is re-expressed in some tumors of the breast and colon. During embryonic development, Cr1 is expressed in inner cell mass cells and the primitive streak, and later is restricted to the developing heart. To investigate the role of Cr1 during mouse development, mice were generated that contain a null mutation of both Cr1 genes, derived from homologous recombination in embryonic stem cells. No homozygous Cr1-/- mice were born, indicating that Cr1 is necessary for embryonic development. Embryos initiated gastrulation and some embryos produced mesoderm up to day E7.5. Increasingly aberrant morphogenesis gave rise to disordered neuroepithelium that failed to produce a recognizable neural tube, or head-fold. Although some biochemical markers of differentiating ectoderm, mesoderm and endoderm were expressed, all the cardiac-specific markers were absent from day E8.7 embryos: (&agr;)MHC, betaMHC, MLC2A, MLC2V and ANF, whereas they were expressed in wild type embryos. The yolk sac and placental tissues continued development in the absence of the embryo until day E9.5 but lacked large yolk sac blood vessels. Chimeric mice were constructed by microinjection of double targeted Cr1(-/- )embryonic stem cells into normal C57BL/6 blastocysts. The Cr1 produced by the normal C57BL/6 cells fully rescued the phenotype of Cr1(-/-) cells, indicating that Cr1 protein acted in a paracrine manner. Cells derived from the embryo proliferated and migrated poorly and had different adhesion properties compared to wild type. Therefore, lethality in the absence of Cr1, likely resulted largely from defective precardiac mesoderm that was unable to differentiate into functional cardiomyocytes. PMID- 9876178 TI - prx-1 functions cooperatively with another paired-related homeobox gene, prx-2, to maintain cell fates within the craniofacial mesenchyme. AB - The paired-related homeobox gene, prx-1, is expressed in the postmigratory cranial mesenchyme of all facial prominences and is required for the formation of proximal first arch derivatives. We introduced lacZ into the prx-1 locus to study the developmental fate of cells destined to express prx-1 in the prx-1 mutant background. lacZ was normally expressed in prx-1(neo); prx-1(lacZ )mutant craniofacial mesenchyme up until 11.5 d.p.c. At later time points, lacZ expression was lost from structures that are defective in the prx-1(neo) mutant mice. A related gene, prx-2, demonstrated overlapping expression with prx-1. To test the idea that prx-1 and prx-2 perform redundant functions, we generated prx 1(neo;)prx-2 compound mutant mice. Double mutant mice had novel phenotypes in which the rostral aspect of the mandible was defective, the mandibular incisor arrested as a single, bud-stage tooth germ and Meckel's cartilage was absent. Expression of two markers for tooth development, pax9 and patched, were downregulated. Using a transgene that marks a subset of prx-1-expressing cells in the craniofacial mesenchyme, we showed that cells within the hyoid arch take on the properties of the first branchial arch. These data suggest that prx-1 and prx 2 coordinately regulate gene expression in cells that contribute to the distal aspects of the mandibular arch mesenchyme and that prx-1 and prx-2 play a role in the maintenance of cell fate within the craniofacial mesenchyme. PMID- 9876179 TI - Defective liver formation and liver cell apoptosis in mice lacking the stress signaling kinase SEK1/MKK4. AB - The stress signaling kinase SEK1/MKK4 is a direct activator of stress-activated protein kinases (SAPKs; also called Jun-N-terminal kinases, JNKs) in response to a variety of cellular stresses, such as changes in osmolarity, metabolic poisons, DNA damage, heat shock or inflammatory cytokines. We have disrupted the sek1 gene in mice using homologous recombination. Sek1(-/- )embryos display severe anemia and die between embryonic day 10.5 (E10.5) and E12.5. Haematopoiesis from yolk sac precursors and vasculogenesis are normal in sek1(-/- )embryos. However, hepatogenesis and liver formation were severely impaired in the mutant embryos and E11.5 and E12.5 sek1(-/- )embryos had greatly reduced numbers of parenchymal hepatocytes. Whereas formation of the primordial liver from the visceral endoderm appeared normal, sek1(-/-) liver cells underwent massive apoptosis. These results provide the first genetic link between stress-responsive kinases and organogenesis in mammals and indicate that SEK1 provides a crucial and specific survival signal for hepatocytes. PMID- 9876180 TI - The novel signal peptides, pedibin and Hym-346, lower positional value thereby enhancing foot formation in hydra. AB - Signaling molecules affecting patterning processes are usually proteins and rarely peptides. Two novel peptides, pedibin and Hym-346, that are closely related to one another have been isolated from Hydra vulgaris and Hydra magnipapillata. Several experiments indicate that both cause a reduction in the positional value gradient, the principle patterning process governing the maintenance of form in the adult hydra. The peptides cause an increase in the rate of foot regeneration following bisection of the body column. Treatment of animals with either peptide for an extended period of time resulted in an apical extension of the range of expression of CnNk-2 along the body column. Such an extension is correlated with a decrease in positional value. Transplantation of tissue treated with Hym-346 results in an increase in the fraction forming feet, and aggregates derived from Hym-346 tissue form more feet and fewer heads. The latter two experiments provide a direct measure of the lowering of positional value in the treated tissue. These results suggest that peptides play signaling roles in patterning processes in cnidaria and, plausibly, in more complex metazoans as well. PMID- 9876181 TI - Mash1 regulates neurogenesis in the ventral telencephalon. AB - Previous studies have shown that mice mutant for the gene Mash1 display severe neuronal losses in the olfactory epithelium and ganglia of the autonomic nervous system, demonstrating a role for Mash1 in development of neuronal lineages in the peripheral nervous system. Here, we have begun to analyse Mash1 function in the central nervous system, focusing our studies on the ventral telencephalon where it is expressed at high levels during neurogenesis. Mash1 mutant mice present a severe loss of progenitors, particularly of neuronal precursors in the subventricular zone of the medial ganglionic eminence. Discrete neuronal populations of the basal ganglia and cerebral cortex are subsequently missing. An analysis of candidate effectors of Mash1 function revealed that the Notch ligands Dll1 and Dll3, and the target of Notch signaling Hes5, fail to be expressed in Mash1 mutant ventral telencephalon. In the lateral ganglionic eminence, loss of Notch signaling activity correlates with premature expression of a number of subventricular zone markers by ventricular zone cells. Therefore, Mash1 is an important regulator of neurogenesis in the ventral telencephalon, where it is required both to specify neuronal precursors and to control the timing of their production. PMID- 9876182 TI - BMP signaling plays a role in visceral endoderm differentiation and cavitation in the early mouse embryo. AB - At E4.0 the inner cell mass of the mouse blastocyst consists of a core of embryonic ectoderm cells surrounded by an outer layer of primitive (extraembryonic) endoderm, which subsequently gives rise to both visceral endoderm and parietal endoderm. Shortly after blastocyst implantation, the solid mass of ectoderm cells is converted by a process known as cavitation into a pseudostratified columnar epithelium surrounding a central cavity. We have previously used two cell lines, which form embryoid bodies that do (PSA1) or do not (S2) cavitate, as an in vitro model system for studying the mechanism of cavitation in the early embryo. We provided evidence that cavitation is the result of both programmed cell death and selective cell survival, and that the process depends on signals from visceral endoderm (Coucouvanis, E. and Martin, G. R. (1995) Cell 83, 279-287). Here we show that Bmp2 and Bmp4 are expressed in PSA1 embryoid bodies and embryos at the stages when visceral endoderm differentiation and cavitation are occurring, and that blocking BMP signaling via expression of a transgene encoding a dominant negative mutant form of BMP receptor IB inhibits expression of the visceral endoderm marker, Hnf4, and prevents cavitation in PSA1 embryoid bodies. Furthermore, we show that addition of BMP protein to cultures of S2 embryoid bodies induces expression of Hnf4 and other visceral endoderm markers and also cavitation. Taken together, these data indicate that BMP signaling is both capable of promoting, and required for differentiation of, visceral endoderm and cavitation of embryoid bodies. Based on these and other data, we propose a model for the role of BMP signaling during peri-implantation stages of mouse embryo development. PMID- 9876183 TI - FGF-7 modulates ureteric bud growth and nephron number in the developing kidney. AB - The importance of proportioning kidney size to body volume was established by clinical studies which demonstrated that in-born defecits of nephron number predispose the kidney to disease. As the kidney develops, the expanding ureteric bud or renal collecting system induces surrounding metanephric mesenchyme to proliferate and differentiate into nephrons. Thus, it is likely that nephron number is related to ureteric bud growth. The expression patterns of mRNAs encoding Fibroblast Growth Factor-7 (FGF-7) and its high affinity receptor suggested that FGF-7 signaling may play a role in regulating ureteric bud growth. To test this hypothesis we examined kidneys from FGF-7-null and wild-type mice. Results of these studies demonstrate that the developing ureteric bud and mature collecting system of FGF-7-null kidneys is markedly smaller than wild type. Furthermore, morphometric analyses indicate that mature FGF-7-null kidneys have 30+/-6% fewer nephrons than wild-type kidneys. In vitro experiments demonstrate that elevated levels of FGF-7 augment ureteric bud growth and increase the number of nephrons that form in rodent metanephric kidney organ cultures. Collectively, these results demonstrate that FGF-7 levels modulate the extent of ureteric bud growth during development and the number of nephrons that eventually form in the kidney. PMID- 9876184 TI - Extrinsic and intrinsic factors control the genesis of amacrine and cone cells in the rat retina. AB - The seven major classes of cells of the vertebrate neural retina are generated from a pool of multipotent progenitor cells. Recent studies suggest a model of retinal development in which both the progenitor cells and the environment change over time (Cepko, C. L., Austin, C. P., Yang, X., Alexiades, M. and Ezzeddine, D. (1996). Proc. Natl. Acad. Sci. USA 93, 589-595). We have utilized a reaggregate culture system to test this model. A labeled population of progenitors from the embryonic rat retina were cultured with an excess of postnatal retinal cells and then assayed for their cell fate choices. We found that the postnatal environment had at least two signals that affected the embryonic cells' choice of fate; one signal inhibited the production of amacrine cells and a second affected the production of cone cells. No increase in cell types generated postnatally was observed. The source of the inhibitor of the amacrine cell fate appeared to be previously generated amacrine cells, suggesting that amacrine cell number is controlled by feedback inhibition. The progenitor cell lost its ability to be inhibited for production of an amacrine cell as it entered M phase of the cell cycle. We suggest that postmitotic cells influence progenitor cell fate decisions, but that they do so in a manner restricted by the intrinsic biases of progenitor cells. PMID- 9876185 TI - A novel role for glycogen synthase kinase-3 in Xenopus development: maintenance of oocyte cell cycle arrest by a beta-catenin-independent mechanism. AB - We have examined the expression of glycogen synthase kinase-3beta in oocytes and early embryos of Xenopus and found that the protein is developmentally regulated. In resting oocytes, GSK-3beta is active and it is inactivated on maturation in response to progesterone. GSK-3beta inactivation is necessary and rate limiting for the cell cycle response to this hormone and the subsequent accumulation of beta-catenin. Overexpression of a dominant negative form of the kinase accelerates maturation, as does inactivation by expression of Xenopus Dishevelled or microinjection of an inactivating antibody. Cell cycle inhibition by GSK-3beta is not mediated by the level of beta-catenin or by a direct effect on either the MAP kinase pathway or translation of mos and cyclin B1. These data indicate a novel role for GSK-3beta in Xenopus development: in addition to controlling specification of the dorsoventral axis in embryos, it mediates cell cycle arrest in oocytes. PMID- 9876186 TI - Wingless signaling in the Drosophila embryo: zygotic requirements and the role of the frizzled genes. AB - Wingless signaling plays a central role during epidermal patterning in Drosophila. We have analyzed zygotic requirements for Wingless signaling in the embryonic ectoderm by generating synthetic deficiencies that uncover more than 99% of the genome. We found no genes required for initial wingless expression, other than previously identified segmentation genes. In contrast, maintenance of wingless expression shows a high degree of zygotic transcriptional requirements. Besides known genes, we have identified at least two additional genomic regions containing new genes involved in Wingless maintenance. We also assayed for the zygotic requirements for Wingless response and found that no single genomic region was required for the cytoplasmic accumulation of Armadillo in the receiving cells. Surprisingly, embryos homozygously deleted for the candidate Wingless receptor, Dfrizzled2, showed a normal Wingless response. However, the Armadillo response to Wingless was strongly reduced in double mutants of both known members of the frizzled family in Drosophila, frizzled and Dfrizzled2. Based on their expression pattern during embryogenesis, different Frizzled receptors may play unique but overlapping roles in development. In particular, we suggest that Frizzled and Dfrizzled2 are both required for Wingless autoregulation, but might be dispensable for late Engrailed maintenance. While Wingless signaling in embryos mutant for frizzled and Dfrizzled2 is affected, Wingless protein is still internalized into cells adjacent to wingless-expressing cells. Incorporation of Wingless protein may therefore involve cell surface molecules in addition to the genetically defined signaling receptors of the frizzled family. PMID- 9876187 TI - Activation of Pax6 depends on somitogenesis in the chick embryo cervical spinal cord. AB - Pax6 is a paired-type homeobox gene expressed in discrete regions of the central nervous system. In the spinal cord of 7- to 10-somite-stage chicken embryos, Pax6 is not detected within the caudal neural plate, but is progressively upregulated in the neuroepithelium neighbouring each newly formed somite. In the present study, we accumulate data suggesting that this initial activation of Pax6 is controlled via the paraxial mesoderm in correlation with somitogenesis. First, we observed that high levels of Pax6 expression occur independently of the presence of SHH-expressing cells when neural plates are maintained in culture in the presence of paraxial mesoderm. Second, grafting a somite caudally under a neural plate that has not yet expressed the gene induces a premature activation of Pax6. Furthermore, after the graft of a somite, a period of incubation corresponding to the individualization of a new somite in the host embryo produces an appreciable activation of Pax6. Conversely, Pax6 expression is delayed under conditions where somitogenesis is retarded, i.e., when the rostral part of the presomitic mesoderm is replaced by the same tissue isolated more caudally. Finally, Pax6 transcripts disappear from the neural tube when a somite is replaced by presomitic mesoderm, suggesting that the somite is also involved in the maintenance of Pax6 expression in the developing spinal cord. All together these observations lead to the proposal that Pax6 activation is triggered by the paraxial mesoderm in phase with somitogenesis in the cervical spinal cord. PMID- 9876188 TI - A gp330/megalin-related protein is required in the major epidermis of Caenorhabditis elegans for completion of molting. AB - A genetic analysis of a gp330/megalin-related protein, LRP-1, has been undertaken in Caenorhabditis elegans. Consistent with megalin's being essential for development of mice, likely null mutations reveal that this large member of the low density lipoprotein receptor family is also essential for growth and development of this nematode. The mutations confer a striking defect, an inability to shed and degrade all of the old cuticle at each of the larval molts. The mutations also cause an arrest of growth usually at the molt from the third to the fourth larval stage. Genetic mosaic analysis suggests that the lrp-1 gene functions in the major epidermal syncytium hyp7, a polarized epithelium that secretes cuticle from its apical surface. Staining of whole mounts with specific monoclonal antibodies reveals that the protein is expressed on the apical surface of hyp7. Sterol starvation can phenocopy the lrp-1 mutations, suggesting that LRP 1 is a receptor for sterols that must be endocytosed by hyp7. These observations indicate that LRP-1 is related to megalin not only structurally but also functionally. PMID- 9876189 TI - Comparison of two immunization schedules with an inactivated hepatitis A vaccine (AvaximTM). AB - BACKGROUND: Inactivated hepatitis A vaccines are licensed with a vaccination schedule based on two injections of vaccine given at least 6 months apart. METHODS: Two vaccination schedules for the inactivated hepatitis A vaccine, AvaximTM (Pasteur Merieux Connaught, Lyon, France), were compared in a monocentric, randomized, open trial. Two doses of the vaccine were given at intervals of either 6 months (0-6 month group) or 12 months (0-12 month group) to 96 adult volunteers. Anti-hepatitis A virus (HAV) antibody titers were determined in a blind fashion using the modified RIA (mRIA) HAVABtrade mark assay. After excluding subjects with positive preimmunization anti-HAV titers and those with protocol deviations, both groups were still comparable by sex ratio and mean age. RESULTS: Four weeks (28 6 4 days) after the first dose, the seroconversion (SC) rate of initially HAV-seronegative subjects (antibody titer < 20 mIU/mL) was 100% in the 0-6 month group and 96. 9% in the 0-12 month group, with corresponding geometric mean titer (GMT) values (95% CI) of 369 mIU/mL (274-497 mIU/mL) and 445 mIU/mL (292-679 mIU/mL), respectively. After 6 months, SC was obtained in all subjects, and the corresponding GMT values were 349 mIU/mL and 359 mIU/mL in the 0-6 month group and the 0-12 month group, respectively. Four weeks after the booster dose given at 6 months, a 14.5-fold rise in GMT was observed. In the 0-12 month group, anti-HAV GMT values decreased by only 20% from 6 months to 12 months with a pre-booster GMT value of 286 mIU/mL at the 12-month evaluation. Four weeks after the booster given at 12 months, a 22. 5-fold rise in GMT was observed. Statistical analysis showed that the two vaccination schedules were comparable in their ability to boost antibody titers. Unsolicited reactions to vaccination were not different to those reported during earlier trials. Less than 12% of the vaccinees reported reactions after the first dose (11/93), or after the booster dose (11/92). CONCLUSIONS: This trial demonstrated antibody persistence is excellent for at least 12 months after one dose of this vaccine, and that a booster may be given at any time between 6 and 12 months after primary immunization. PMID- 9876190 TI - Risk of malaria in visitors to the Kruger National Park, South Africa. AB - BACKGROUND: The risk of malaria to visitors to South Africa's nature reserves is unknown. Current recommendations for travelers are based on malaria incidence rates in local communities. These rates may well overestimate travelers' malaria risk and lead to unnecessary use of antimalarial prophylaxis with associated drug related adverse events. This study investigated the incidence of malaria in visitors to South Africa's Kruger National Park (KNP). METHODS: Postal questionnaires were sent to the cohort of visitors staying in the KNP during April 1996, 6 to 8 weeks after they returned to their homes. Nonrespondents received duplicate questionnaires 8 weeks later. Responses from 3267 groups, representing 11,107 visitors (56. 8%) allowed determination of the malaria attack rate in this group of travelers and assessment of associations between malaria risk and a number of factors, including type of accommodation used during the visit; duration of stay; use of chemoprophylaxis; and use of personal protection measures. All travelers indicating that they had suffered an episode of malaria following their visit were telephonically contacted and their health providers traced to determine the basis of diagnosis and clinical outcome. RESULTS: One clinically suspected and four confirmed cases of Plasmodium falciparum malaria among the visitors suggest a low attack rate of 4. 5 cases per 10,000 visitors during April, the highest risk month for malaria in South Africa. No association was found between malaria risk and accommodation type, duration of stay, use and type of chemoprophylaxis, and use and number of personal protection measures. CONCLUSIONS: These findings confirm the importance of careful assessment of local malaria risk before individualized advice is provided to travelers. PMID- 9876191 TI - Health hazards in international tourists visiting Paris in August: a five-year retrospective epidemiologic survey. AB - BACKGROUND: Travel-related illnesses have been studied in visitors to developing countries, but no studies have examined the incidence of health problems in visitors to developed countries. METHODS: 4, 093 foreign tourists visiting Paris in August and attending to emergency medical care for acute health problems were included in an epidemiological survey conducted over 5 consecutive years. The objective was to determine what types of acute health problems occur in a foreign tourist population and to estimate the incidence of the main health hazards. RESULTS: Gastroenteritis represented the main cause of medical care in that population (from 14.5-21.9%) followed by traumatology, ENT problem, viral syndrome and dermatology which represented altogether 60-64% of all medical problems. Two factors were related to the distribution of diseases observed: age and nationality. The monthly incidence of gastroenteritis was estimated to be between 1.33 to 2.92 per 10,000 visitors, and the overall incidence of health problems between 8 to 10 per 10,000. CONCLUSIONS: Even if the incidence rate of gastroenteritis is low compared with developing countries, further studies are needed to support the hypothesis that gastroenteritis could be attributed to sanitary conditions in some restaurants of the French capital. PMID- 9876192 TI - Health problems of corporate travelers: risk factors and management. AB - BACKGROUND: Numerous studies have been done regarding health problems experienced by tourists in developing countries; however, little data exist about these health risks and illnesses experienced by corporate travelers. METHODS: The authors examined by electronic survey the health risks encountered, compliance with pretravel health recommendations, and illnesses and injuries experienced by employees of the Coca-Cola Company who travel internationally. RESULTS: Two hundred twenty-six travelers responded. Although most travelers ate meals at their hotels and chose foods that were cooked and still hot, over half also ate foods that remained at room temperature for prolonged periods and/or ate from cold salad bars. Almost half drank untreated tap water. Thirty-five percent of travelers developed diarrhea and 29% reported respiratory illnesses, with 12% seeking medical attention for their problems. Forty-three percent of those traveling to malarious regions admitted to noncompliance with antimalarial recommendations. Health kits provided were used by only 51% of travelers. CONCLUSIONS: Although many corporate travelers followed pretravel health recommendations, some did not. Injuries, fever, and illnesses such as diarrhea and respiratory infections occurred. Strategies to improve access to the travel clinic and the acquisition of health information and travel health kits are being implemented. The health risks and behaviors of corporate travelers, including the potential impact of psychosocial stressors, need greater attention. PMID- 9876193 TI - Effectiveness of personal protection measures against mosquito bites for malaria prophylaxis in travelers. AB - BACKGROUND: To determine the effectiveness of personal protection measures (PPM) against mosquito bites in travelers as tools of malaria prevention. METHODS: Two step questionnaire survey conducted in European tourists who visited East Africa: cross-sectional evaluation during the return flight with follow-up 12 weeks after return home. Among 100,336 tourists participating in a malaria survey, 89,617 had answered all questions relating to PPM and to malaria occurrence. The main outcome measure was the occurrence of malaria. RESULTS: Only 2% of tourists visiting East Africa regularly take all four of the most important PPM. Regular use of PPM resulted in a small, but significant reduction of malaria incidence when travelers were interviewed 12 weeks after returning home. Air-conditioned rooms and clothing which covered arms and legs were effective, whereas repellants, insecticides, coils, etc. showed no significant effect. CONCLUSIONS: In general, advice relating to all possible PPM in travelers should be concentrated on those who are at greatest risk of bites, and thus of malaria. For those at low and intermediate risk, preference should be given to convenient measures with proven effectiveness, such as using air-conditioned rooms and appropriate clothing. PMID- 9876194 TI - Retrospective study of malaria cases treated in Newcastle General Hospital between 1990 and 1996. AB - BACKGROUND: Malaria, in particular Falciparum malaria, continues to pose a substantial risk to travelers to endemic areas. METHODS: In this study we examined 93 case notes of patients with malaria treated in our department between 1990 and 1996. RESULTS: Forty-seven (50.5%) patients had infection with Plasmodium falciparum, 41 (44.1%) had Plasmodium vivax and 5 (5.4%) had Plasmodium ovale. One of these patients had a dual infection with P. falciparum and P. vivax. None of our patients had Plasmodium malariae. Forty-four of the P. falciparum cases (93.6%) were imported from sub-Saharan Africa, 33 of the P. vivax cases (78.5%) were imported from the Indian subcontinent. All the P. ovale cases were imported from sub-Saharan Africa. Fifty-four of our patients (58.1%) did not take any form of chemoprophylaxis. Forty-two out of 93 (45.2%) of the "travelers" were settled immigrants in the UK. Seventy-eight percent of travelers of British caucasian origin took prophylaxis whereas only 13.5% of travelers of ethnic minorities origin took prophylaxis. CONCLUSIONS: Greater awareness of the risk of malaria by travelers and medical practitioners in UK must be encouraged and in particular appropriate chemoprophylaxis instituted for travelers to chloroquine-resistant areas. PMID- 9876195 TI - Anxiety and health problems related to air travel. AB - BACKGROUND: A significant proportion of air travelers experience situational anxiety and physical health problems. Take-off and landing are assumed to be stressful, but anxiety related to other aspects of the air travel process, anxiety coping strategies, and in-flight health problems have not previously been investigated. METHODS: We aimed to investigate frequency of perceived anxiety at procedural stages of air travel, individual strategies used to reduce such anxiety, and frequency of health problems on short-haul and long-haul flights. A questionnaire measuring the occurrence and frequency of the above was administered to two samples of intending travelers during a 3 month period to: (a) 138 travel agency clients, and (b) 100 individuals attending a hospital travel clinic. RESULTS: Of the 238 respondents, two thirds were women. Take-off and landing were a perceived source of anxiety for about 40% of respondents, flight delays for over 50%, and customs and baggage reclaim for a third of individuals. Most frequent anxiety-reduction methods included alcohol and cigarette use, and distraction or relaxation techniques. Physical health problems related to air travel were common, and there was a strong relationship between such problems and frequency of anxiety. Travel agency clients reported more anxiety but not more physical health symptoms overall than travel clinic clients. Women reported greater air-travel anxiety, and more somatic symptoms than men. CONCLUSIONS: Significant numbers of air travelers report perceived anxiety related to aspects of travel, and this is associated with health problems during flights. Airlines and travel companies could institute specific measures, including improved information and communication, to reassure clients and thereby diminish anxiety during stages of air-travel. Medical practitioners and travel agencies should also be aware of the potential stresses of air travel and the need for additional information and advice. PMID- 9876196 TI - Are preventive HIV interventions at airports effective? AB - BACKGROUND: Few empirical data exist on the impact of preventive human immunodeficiency virus (HIV) interventions on intended and actual sexual behavior of international tourists. The present cross-sectional study is based on a 2 3 2 design. METHODS: The sample consisted of departing and arriving passengers (n = 3100) at Zurich Airport with destinations in countries where heterosexual HIV transmission is dominant. While 41% of the tourists obtained information about safer sex, the remaining 59% without such intervention served as control group. Departing passengers completed a short questionnaire focusing on their planned sexual behavior. Arriving passengers were asked about their actual behavior during the journey. Subjects of the intervention group also evaluated the impact of the consultation. RESULTS: Most travelers appreciated the intervention and reported that they received important information. Members of the intervention group were better informed than those of the control group about the risk of heterosexually transmitted HIV infection (p <.01). They also indicated more often that they could imagine having casual sex abroad (23% vs 16%, p <.01). However, the two groups did not differ with regard to planned condom use or actual sexual behavior. Whereas most of departing passengers indicated that they would use condoms consistently, only half of the passengers who reported casual sex actually did so. Subjects who refused to participate in the intervention tended to consider it as irrelevant and reported less consistent condom use. CONCLUSIONS: Although travel health interventions focusing on casual sex are appreciated and increase the knowledge, they failed to result in significant behavior modification. Future projects should attempt to approach possible risk groups more specifically and to have more impact. PMID- 9876197 TI - Health aspects of Antarctic tourism. AB - Increasing numbers of seaborne tourists are visiting Antarctica, with most coming from the United States (3503 in 1996-97), Germany (777), and Australia (680; cf. 356 in 1994-95 and 410 in 1995-96). The impression among travel medicine clinicians is that, each year, more prospective travelers seek advice about the health demands of this type of adventure, mostly relating to fitness for travel, exposure to extreme cold, hazards in ice and snow, and other potential health risks. This is a recent phenomenon. While a regular shipping service had been established between the Falklands and the subantarctic islands of South Georgia and the South Shetlands by 1924, the first documented tourists accompanied an Argentine expedition to the South Orkneys in 1933.1 Commercial airline flights over these islands and the Antarctic Peninsula began in 1956, from Chile, and recreational cruises to the Peninsula began in 1958. Tourist numbers subsequently grew slowly, for what was clearly an exclusive and very expensive undertaking, with few ships available for these hazardous voyages. From 1957 to 1993, 37,000 tourists visited by sea, most seeing only the Peninsula.2 The dramatic recent growth in numbers is a consequence of the collapse of the Soviet Union. The small fleet of ice-strengthened research vessels and working icebreakers, which was made redundant by withdrawal of central government support from isolated communities and military activities along the northern coast of Siberia (and from Antarctic research bases), now accounts for the bulk of charter-cruise tourism to Antarctica, at competitive prices. According to the International Association of Antarctica Tour Operators,3 7322 people traveled to Antarctica on commercially organized voyages in the 1996-97 season, and a record 10,000 shipborne visitors were expected for the 1997-98 season (November-March), traveling mainly from South America to the Peninsula on 15 ice-reinforced vessels, each carrying between 36 and 180 passengers. Most tours embark from Ushuaia in southern Argentina, or Punta Arenas in Chile, although occasional cruises leave from Australia, New Zealand, or even South Africa. PMID- 9876198 TI - Dangers in the ocean: the traveler and marine envenomation. II. Marine vertebrates. AB - Envenomation both from jellyfish and other marine animals causes human deaths and severe morbidity in many countries in the world having tropical, or subtropical waters. In part II the world distribution of venomous marine vertebrates is discussed, together with simple first aid and effective medical treatment. Suggestions are made for the awareness and prevention of marine envenomation. With travel becoming more popular than ever, general practitioners and travel medicine consultants must routinely advise their patients on the worldwide hazards of marine envenomation. PMID- 9876199 TI - Deployed US Army soldiers' knowledge and use of personal protection measures to prevent arthropod-related casualties. AB - The recommendation to use personal protection measures (PPMs) to prevent arthropod-related diseases and nuisance bites is a common element of travel medicine consultation.1-3 Prevention of arthropod-related casualties is especially important to the military, given the often intense exposure of service members to biting arthropods and the threat of personnel losses to mission success. In the 1980s, 75% deet (N,N-diethyl-m-toluamide) was the US military issue insect repellent for use on skin and clothing. Collaboration between military and civilian researchers led to the implementation in 1991 of the current US military system of PPMs which has three components: topical application of 33% extended-duration deet, treatment of field uniforms with permethrin, and proper wearing of field uniforms.4-6 Compared to military-issue 75% topical deet, 33% extended-duration deet prevents bites up to three times longer (as long as 12 hours), is less greasy, and has lower plasticizing properties. Field uniforms treated with the contact toxicant, permethrin, are also necessary to minimize bites from crawling arthropods such as ticks and chiggers. Implementation of all three components of this system is a safe and effective means of reducing the threat posed by biting arthropods.7 We conducted a questionnaire survey to assess the degree of deployed soldiers' knowledge of the US military's system of PPMs and use of PPMs in general. Survey results may promote the development of better ways to advise and teach military and civilian travelers about the proper use of PPMs given the multitude of available products and practices. PMID- 9876200 TI - Fatal grand mal seizure in a Dutch trekker. AB - A 35-year-old healthy Dutch woman went on a trek (Lang Tang) in Nepal up to an approximate altitude of about 3800 meters. She had no prior history of any medical problems except attacks of generalized epilepsy when she was 19 years old, which had been controlled with antiepileptic medications. She had had no attacks after the age of 20. A CT scan done around that time had apparently been normal. On this trek she had developed diarrhea which had been cured with norfloxacin 400 mg two times per day for 3 days. Two days later, while descending, she developed a grand mal seizure at an altitude of 3300 meters, after which she developed a classic postictal phase but gradually recovered. She developed grand mal seizures again the next day, but when she went to a travel clinic in Kathmandu, she had been seizure free for 72 hours. She also revealed that she had not suffered from acute mountain sickness on the trek. She also had a prior history of gastroenteritis at high altitude which improved significantly with norfloxacin, a quinolone antibiotic. This was corroborated by her party. Upon examination she was fully conscious and oriented to person, place and time. Her pulse was 70 beats per minute and her BP was 110/80 mm of Hg. Her fundi and cranial nerves exam were completely normal. Her abstract thinking, gait, power, tone, reflexes and other facets of her neurologic exam revealed absolutely no abnormalities. Her cardiovascular exam revealed a normal rhythm with no murmurs or bruits. The rest of her exam was also normal. She revealed that she played tennis on a regular basis and was an outdoor person. She had not trekked before in the Himalayas and it was uncertain if she had been to high altitude before. She was on no medications at the time. There was no history of drug abuse. She did not smoke and consumed few alcoholic beverages. She had not consumed any alcohol on the trek. She was advised to get a CT scan (she declined as she was going home to Holland in 2 days) and she was prescribed a loading dose of phenytoin 1 g orally spread over several hours and it was recommended she take 300 mg of phenytoin per day. She was going to see her neurologist in Holland on arrival there. She went to her hotel in Kathmandu while her friends went to fill the prescription of phenytoin. When her friends returned to the hotel she was having another grand mal seizure. Medical help was sought, but she died before the doctor arrived to control her seizures. When the doctor did arrive and carried out CPR for half an hour it was to no avail as she continued to have no pulse or blood pressure. PMID- 9876202 TI - Critical incident stress debriefing in international aid workers. AB - Missionaries are well known to suffer the effects of stress. Patricia Miersma relates missionary stress to combat related stress. 1 Development workers too are known to be at increased risk of death whilst overseas-mostly due to traumatic incidents. Relief workers voluntarily enter high stress situations. These overseas workers are at real risk of Post Traumatic Stress Disorder (PTSD). The issue of Critical Incident Stress Debriefing (CISD, or Psychological Debriefing) has been critically examined in an editorial in the British Medical Journal.3 The first randomized, controlled study of CISD that we are aware of (for motor vehicle accident survivors) was published in 1996.4 With 54 intervention subjects, it did not demonstrate effectiveness. PMID- 9876201 TI - Cutaneous larva migrans, sacroileitis, and optic neuritis caused by an unidentified organism acquired in Thailand. AB - We report the case of a 32-year-old pregnant woman with an unidentified intraocular parasite. The parasite, which had been acquired in Thailand, caused cutaneous larva migrans, sacroileitis, and 2 years later optic neuritis and panuveitis. The patient was successfully treated with ivermectin and albendazole. The diagnostic possibilities of this peculiar presentation are discussed. Parasitic infections are a leading cause of medical problems in travelers to tropical countries.1 While most parasites cause gastrointestinal problems, some may migrate throughout the body and lodge in critical organs. Ocular parasitic infections may occur by direct inoculation onto the eye,2 or incidentally during systemic migration. Subconjunctival parasites are easily diagnosed by removal and careful microscopic examination.3 Parasites, which lodge within the eye, are more difficult to diagnose, especially if not removed. In this report we describe a patient who presented with an intraocular parasite causing optic neuritis and panuveitis, 2 years after travel to Thailand. PMID- 9876204 TI - Prevalence of antimicrobial resistance in eighty clinical isolates of Helicobacter pylori. AB - We studied the in vitro susceptibility of 80 Helicobacter pylori clinical isolates to several antimicrobials commonly used to treat the infection, using the Epsilometer test (E-test). We also compared E-test and disk diffusion test in determining H. pylori susceptibility to the same antibiotics. We found a high prevalence of H. pylori strains resistant to metronidazole (23.7%), whereas the prevalence of H. pylori strains resistant to clarithromycin was 10%. Also, a significant correlation was found between MICs obtained with the disk diffusion test and E-test for metronidazole and clarithromycin. In conclusion, our study confirms a high prevalence of metronidazole- or clarithromycin-resistant H. pylori strains. Also, our data suggest that the E-test is a single, reliable, and cost-effective method to assess in vitro susceptibility of H. pylori to antimicrobial agents commonly used to eradicate the infection. PMID- 9876203 TI - Pharmacokinetics of cefodizime in patients with various degrees of renal failure. AB - The pharmacokinetics of cefodizime, a new expanded-spectrum cephalosporin for parenteral use, was studied in 45 subjects with various degrees of renal failure. Patients were divided into five groups according to the following creatinine clearances: group I >80 ml/min; group II <80-30 ml/min; group III <30-15 ml/min; group IV <15-5 ml/min and group V <5 ml/min. Cefodizime was administered as a 1 g i.v. bolus. Plasma and urinary concentrations of cefodizime were determined by high-performance liquid chromatography, using for detection UV absorbance. The following pharmacokinetic parameters were calculated: maximum plasma concentration (C5 min), area under the plasma concentration-time curve (AUC), terminal half-life (T1/2), terminal rate constant (lambda-z), total clearance (Clt), volume of distribution (Vd), mean residence time (MRT), urine data-derived terminal half-life (T1/2 r), renal clearance (Clr). The results of this study showed that renal failure induced changes in cefodizime pharmacokinetics. Our data demonstrated a close correlation between degree of renal impairment and pharmacokinetic changes. The maximum plasma concentration (C5 min) was higher in patients with renal failure; T1/2 was increased; AUC also increased from 470.40 +/- 17.80 mg.h/l in the control group to 1,562.30 +/- 170.8 mg.h/l in group V. Moreover, no side effect was observed after treatment with 1 g i.v. of cefodizime. Although renal failure induces significant changes of cefodizime pharmacokinetics, the drug was well tolerated and only in patients with severe renal insufficiency we advise to monitor the interval dose of cefodizime or adjust doses to renal function. PMID- 9876206 TI - In vitro activity of trovafloxacin (CP 99,219), a new fluoroquinolone against hospital isolates. AB - The susceptibility of 492 Enterobacteriaceae, 227 other gram-negative bacteria, 448 gram-positive bacteria and 108 anaerobic organisms was determined by the agar dilution method against trovafloxacin and other antibiotics. Trovafloxacin was highly active against most of the Enterobacteriaceae including Enterobacter spp. and Citrobacter spp. [minimum inhibitory concentration (MIC)90 <1 mg/l], Acinetobacter spp. and Pseudomonas aeruginosa (MIC90 = 0.25 and 2 mg/l, respectively). The antimicrobial activity was extended to the gram-positive bacteria including streptococci, Streptococcus pneumoniae, coagulase-negative staphylococci and methicillin-sensitive Staphylococcus aureus with MIC90 <1 mg/l. Enterococci and methicillin-resistant S. aureus were inhibited (MIC90 = 2 mg/l; sparfloxacin and ciprofloxacin were 16 and 64 mg/l, respectively). Almost all anaerobic organisms were inhibited by trovafloxacin (MIC90 = 1 mg/l). PMID- 9876205 TI - Increasing prevalence of ampicillin- resistant, non-beta-lactamase-producing strains of Haemophilus influenzae in children in Japan. AB - Among Haemophilus influenzae isolated from children with respiratory tract infections, the evolution of ampicillin resistance was investigated during 1996 and 1997 in Japan. beta-Lactamase production was assessed and minimum inhibitory concentrations (MICs) of eight antimicrobial agents were determined using a broth microdilution method in Mueller-Hinton-lysed horse blood medium. Of 74 H. influenzae, 11 strains (14.9%) produce beta-lactamase and were thus highly resistant to ampicillin (MIC of >4.0 microgram/ ml). In addition, moderate resistance to ampicillin, defined as an MIC of >==1.0 microgram/ml, was noted in 44.4% of all beta-lactamase-negative isolates. These beta-lactamase-negative ampicillin-resistant (BLNAR) organisms were resistant to other cephalosporins such as cefpodoxime and cefdinir, while beta-lactamase-producing strains were susceptible to them. Cefditoren, cefteram, and minocycline were active against all strains studied, whereas cefaclor and clarithromycin were inactive against all H. influenzae isolates in this study. Results indicate that BLNAR strains have emerged among children with respiratory tract infections in Japan. PMID- 9876207 TI - Effect of the growth rate of Pseudomonas aeruginosa biofilms on the susceptibility to antimicrobial agents: beta-lactams and fluoroquinolones. AB - The growth rate of biofilm bacteria of a leucine-requiring mutant Pseudomonas aeruginosa HU1 was regulated by the leucine concentration in a chemically-defined medium. The semiquantitative measurement of glycocalyx and scanning electron microscopy revealed that the kinetics of HU1-biofilm formation were dependent on the incubation time and the leucine concentration in the medium. The effect of the growth rate of biofilm cells on their susceptibility to antimicrobial agents, three beta-lactams and four fluoroquinolones, was evaluated. beta-Lactams showed weak bactericidal activity to biofilm cells; the activity was greater in younger biofilm cells growing in high concentrations of leucine. Fluoroquinolones revealed strong bactericidal activity to biofilm bacteria regardless of the growth rate. The following is suggested: the bactericidal action of beta-lactams against biofilm cells is affected by the cell growth rate, while that of fluoroquinolones is considerably greater and independent on the growth rate. PMID- 9876208 TI - Role of inhibition of penicillin binding proteins and cell wall cross-linking by beta-lactam antibiotics in low- and high-level methicillin resistance of Staphylococcus aureus. AB - We have investigated the relationships between the minimum inhibitory concentrations (MICs) of eight beta-lactams and their binding affinities of penicillin binding proteins in low- and high-level methicillin-resistant Staphylococcus aureus (MRSA). For high-level MRSA (H-MRSA), a significant correlation was found between IC50 for PBP2A and MIC, whilst no clear relationship was apparent for low-level MRSA (L-MRSA). However, the product of the IC50s for PBP2A and PBP4 significantly correlated with the MIC in L-MRSA. Since PBP4 is thought to mediate secondary cross-linking of the staphylococcal cell wall, we analyzed the effect on cell wall cross-linking of L- and H-MRSA of two representative drugs (cefoselis and flomoxef). Interestingly, the decrease in cell wall cross-linking, which was clearly observed in L-MRSA, was diminished with H-MRSA. It is concluded that for L-MRSA, a reduction in cell wall cross linking caused by inhibiton of PBP4 may contribute to the antimicrobial activity of beta-lactam antibiotics, while for H-MRSA, inhibition of PBP2A mainly determines the antimicrobial activity. Since neither alteration of expression nor binding affinity of PBP4 were observed in these studies, unknown factors must operate to diminish the effect of PBP4 inhibition and contribute to the mechanism of high level resistance of MRSA. PMID- 9876209 TI - A new fluoroquinolone derivative exhibits inhibitory activity against human immunodeficiency virus type 1 replication. AB - The inhibitory activity of several fluoroquinolone antibiotics against human immunodeficiency virus type 1 (HIV-1) replication was investigated. R-71762, (+/ ) 9-fluoro-3-fluoromethyl-2, 3-dihydro-10-[4-(2-pyridyl)-1-piperazinyl]-7-oxo-7H pyrido[1,2, 3-de][1,4]benzoxazine-6-carboxylic acid, protected MT-4 cells from HIV-1-induced cytopathic effects. Furthermore, the compound inhibited virus replication both in acutely and in chronically HIV-1-infected cells. On the other hand, ofloxacin, levofloxacin, ciprofloxacin, norfloxacin and enoxacin, that were previously reported to be protective against HIV-1-induced cytopathic effects, did not show any protective activity in our assay system. These results indicate that R-71762 is a novel inhibitor of HIV-1 replication and is effective even in HIV-1 chronically infected cells. PMID- 9876210 TI - Reduction of mucoid Staphylococcus epidermidis adherence to intraocular lenses by selected antimicrobial agents. AB - The effect of low concentrations of various antimicrobial agents on the adherence of mucoid Staphylococcus epidermidis to intraocular lenses was investigated. Adherent growth of S. epidermidis on to polymethylmethacrylate lenses was greatly diminished by ciprofloxacin and clindamycin (>75%), followed by ceftazidime ( approximately 50%) and less affected by gentamicin and vancomycin (<25%) compared to the adherent growth of control untreated cells. The reduction in adherence growth was concentration-dependent and found to be due to the inhibition of slime production since no growth inhibition was observed at the concentrations used. Cell surface hydrophobicity was markedly reduced in parallel to the inhibition of adherence as compared to the control cells. The differential effect of antibiotics on adherence, regardless of their antimicrobial activity, may have a clinical significance in reducing the incidence of postoperative endophthalmitis and intraocular inflammation due to the reduction of cell adherence. PMID- 9876211 TI - Enhanced cytogenetic and antineoplastic effects by the combined action of two esteric steroidal derivatives of nitrogen mustards. AB - The authors studied the effect of two modified steroids containing different proportions (%) of alkylating agents alone or in combination on sister chromatid exchange (SCE) rates and on human lymphocyte proliferation kinetics. The antitumor activity of these compounds was tested on leukemia P388- and leukemia L1210-bearing mice. The two chemicals in mixtures enhance SCE induction and antitumor activity in a synergistic manner. The homo-aza-steroidal ester of p bis(2-chloroethyl)aminophenyl acetic acid was found to be more effective than the homo-aza-steroidal ester of o-bis(2-chloroethyl)aminobenzoic acid in causing cytogenetic damage and antineoplastic activity. A correlation was observed between the magnitude of the SCE response and the depression of the cell proliferation index. The order of the antitumor effectiveness of the five different treatments tested coincided with the order of the cytogenetic effects they induced. PMID- 9876212 TI - Treatment of malaria tropica with a fixed combination of rifampicin, co trimoxazole and isoniazid: a clinical study. AB - The treatment of malaria tropica is becoming difficult because of the increasing drug resistance rates of Plasmodium falciparum against several of the currently available antimalarias. A fixed combination of rifampicin, co-trimoxazole and isoniazid (CotrifazidTM, CF) was found to be highly effective for the treatment of malaria tropica. The aim of the present study in Kenya was to scrutinize this finding in a 14-day trial. Patients with malaria tropica were given in an open, double-arm randomized study CF for 5 days, or chloroquine or pyrimethamine/sulphadoxine as the control. Because of an apparently better activity and tolerance of CF, the randomization had to be stopped after the enrollment of 50 patients. A total of 61 patients in both groups (35 of them between 2 months and 6 years of age) were available for final analysis. All 41 patients treated with CF, originally positive in their blood smears, turned negative; in 2 cases blood smear positivity reappeared on day 14. There were 7 failures in the control group, 4 of them a primary one. Four of those failures were turned negative with CF, 2 failed with CF also, and 1 disappeared. The tolerance of CF was excellent even in infants. In our experience, CF is very well suited for the treatment of malaria tropica, also in cases of apparent drug resistance of P. falciparum against other antimalarials, and even in severe cases of the disease. PMID- 9876213 TI - Strategies that support declining cholinergic neurotransmission in Alzheimer's disease patients. AB - With an increased public awareness of Alzheimer's disease (AD), the last two decades have witnessed an immense research effort directed towards discovering the cause of AD with the ultimate hope of developing safe and effective pharmacological treatments. Biochemical and histopathological changes of neurotransmitter markers in the brains of AD patients both at postmortem and neurosurgical cerebral biopsy have demonstrated an association between a decline in learning and memory, and a deficit in cholinergic and associated excitatory amino acid neurotransmission. These observations illustrate the selective neurotransmitter pathology of AD. Accordingly, although there is presently no 'cure' for AD, a large number of potential therapeutic strategies have emerged that are designed to correct the loss of cholinergic neurotransmission in AD. Such strategies include increasing acetylcholine synthesis, enhancing its presynaptic synthesis and/or release, potentiating its pre- or postsynaptic action, obstructing its metabolism, or by influencing the function of cholinergic neurones themselves by administration of nerve growth factor or by trans-synaptic modulation. Indeed, the cholinesterase (ChE) inhibitors have gained the most attention. Adverse events limited the usage of the first ChE inhibitors, but more recently introduced, well-tolerated compounds, for example donepezil, have confirmed efficacy in delaying the deterioration of symptoms of AD, a valuable treatment target considering the progressive nature of the disease. Other agents which affect the cholinergic system, directly or indirectly, that are in the early stages of development for the treatment of AD are also discussed. PMID- 9876215 TI - Clinical profile of donepezil in the treatment of Alzheimer's disease. AB - Although the underlying pathogenesis of Alzheimer's disease (AD) is not fully understood, one of its key features is the widespread loss of central cholinergic innervation, known to be fundamental for cognitive processes. This finding led to the hypothesis that pharmacological enhancement of acetylcholine (ACh) neurotransmission may alleviate the symptoms of AD. Currently, cholinergic therapy, particularly cholinesterase (ChE) inhibition, represents the most realistic approach to the symptomatic treatment of AD. Donepezil HCl, for example, is a piperidine-based, reversible acetylcholinesterase (AChE) inhibitor, chemically distinct from other ChE inhibitors and rationally designed for the symptomatic treatment of AD. It is highly selective for centrally acting AChE, with little or no affinity for butyrylcholinesterase, present predominantly in the periphery. Phase I and II clinical trials demonstrated donepezil's favourable pharmacokinetic, pharmacodynamic and safety profile with no requirement for dose modification in the elderly or in patients with renal or hepatic impairment. Furthermore, its long half-life supports a simple and convenient once-daily dosing regimen. Subsequent to encouraging phase II clinical trial results, two pivotal, randomized, double-blind phase III trials (of 15 and 30 weeks' duration) demonstrated highly significant improvements in cognition and global function in mild to moderately severe AD patients treated with either 5 or 10 mg/day donepezil compared with placebo. Adverse events in the phase II and III trials, primarily cholinergic in nature, were transient and generally mild in severity and resolved during continued donepezil administration. Thus, the donepezil clinical trials programme has shown that this drug is a clinically effective and well-tolerated, once-daily treatment for the symptoms of mild to moderately severe AD. PMID- 9876214 TI - Neuropsychiatric symptoms and cholinergic therapy for Alzheimer's disease. AB - Neuropsychiatric abnormalities, as well as the commonly associated neuropsychological symptoms, are clinical characteristics of Alzheimer's disease (AD), the most common form of dementia. Thus, in addition to a general cognitive and functional decline, neuropsychiatric manifestations, such as agitation, apathy, anxiety, psychoses and disinhibition, are frequently evident in AD patients. Such neuropsychiatric symptoms of AD are the source of considerable patient and caregiver distress, resulting in the prescription of neuroleptics, benzodiazepines or other psychotropic agents, and are a major factor in the decision to transfer the care of patients into nursing homes. Recent evidence suggests that some neuropsychiatric changes associated with AD are related to the cholinergic deficits in the brains of AD patients and that such abnormalities may be responsive to cholinergic therapy. Cholinergic drug therapies indicated for the symptomatic treatment of AD, for example tacrine and the newer cholinesterase (ChE) inhibitors such as donepezil, have been demonstrated to improve memory, language and praxis. Furthermore, although less is known about the effect of ChE inhibitors on the neuropsychiatric symptoms of AD, preliminary evidence suggests that they reduce apathy, anxiety, hallucinations, disinhibition and aberrant motor behaviour. Thus, the newer-generation ChE inhibitors that are well tolerated, easy to administer and show promise in reducing the cognitive, as well as neuropsychiatric disturbances of AD, may emerge as important treatments for some neuropsychiatric symptoms in patients with central cholinergic deficits, including AD. PMID- 9876216 TI - Managing expectations in the long-term treatment of Alzheimer's disease. AB - Alzheimer's disease (AD) is rare before the sixth decade of life, but becomes more prevalent with age. As the elderly proportion of the population is increasing, the prevalence of AD is predicted to expand. Although there is no 'cure' for AD, focused therapeutic interventions may be initiated at a treatable stage of the disease if a diagnosis of AD is established early. For example, cholinesterase inhibitor therapy that is targeted towards the mild to moderate disease stages might be initiated. Results from an open-label study suggest that the benefits of donepezil are maintained for at least 2 years. The disease has major implications for the patients' caregivers, who experience a burden that increases over time and which may result in a significant loss of income. Both patients and caregivers can benefit from pharmacological and nonpharmacological therapeutic strategies. For example, caregiver support programmes may ease the burden of AD and result in a greater potential for the successful treatment of patients with AD, and may delay or even prevent the early institutionalization of the patient. Currently, therefore, pharmacological treatments, supported by nonpharmacological strategies, are the mainstay of therapy for AD patients and their caregivers. In the future, studies are required to clarify whether other compounds, including oestrogen replacement therapy, antioxidants and anti inflammatory, may slow or delay disease onset, or halt disease progression. PMID- 9876217 TI - Emerging roles for proteinases in cancer. AB - Metalloproteinases and serine proteinases have been associated with tumor invasion and formation of metastasis which represent the major obstacles to cancer cure. The contribution of proteinases in these processes was initially thought to be the destruction of extracellular matrices. However, recent evidence suggests that they mainly affect tumor growth rather than invasion. Proteinases can indeed generate active matrix protein fragments, influence the release, the activation and the bioavailability of growth factors, and consequently modulate tumor cell growth, apoptosis and angiogenesis. Additionally, proteinases, their receptors and/or inhibitors can be directly involved in cell migration and in the processing or shedding of cell surface proteins. Further elucidation of the functions of proteinases is essential for the development of novel anticancer strategies. PMID- 9876219 TI - Oral squamous cell carcinoma invasion is associated with a laminin-5 matrix re organization but independent of basement membrane and hemidesmosome formation. clues from an in vitro invasion model. AB - The basement membrane (BM) molecule laminin-5 represents the major component of the anchoring filaments, the BM counterpart of the hemidesmosomes. Because laminin-5 is abundantly deposited adjacent to budding invasive carcinoma cells in vivo, a special invasion-promoting role is suspected. We present a 3D collagen type I gel invasion model for oral squamous cell carcinoma (OSCC) using the newly established OSCC cell lines PE/CA-PJ15, PE/CA-PJ34, and PE/CA-PJ41. The carcinoma cells on the surface of the collagen type I gel show an invasive growth exclusively in the presence of gel-inoculated (myo)fibroblasts yielded from nodular palmar fibromatosis or tumour stroma. The model is characterized by an immunohistochemical and ultrastrcutural failure of structural BM and hemidesmosome formation. In the model an invasion-associated modulation of the laminin-5 deposition (alpha3, clone BM165; gamma2, clone GB3) was demonstrated: in invasive areas a strong ribbon-like immunostaining in the tumor-gel interface. The results obtained from the invasion model suggest that the assumed invasion promoting ability of the BM molecule laminin-5 is realized independent of a structural BM and hemidesmosome formation. PMID- 9876218 TI - Spontaneous metastasis of rat liver epithelial cells transformed with v-raf and v raf/v-myc: association with different phenotypic properties. AB - Cloned v-raf, v-raf/v-myc, and spontaneously transformed rat liver epithelial (RLE) cell lines were examined for meastatic capability in nude mice, using the LacZ gene as a marker for quantitation of micrometastases. Six cloned lines (R3611-T lines) derived from nude mouse xenografts of the v-raf transformed R3611 3 cells displayed variable metastatic capabilities. Three of six subcutaneously inoculated R3611-TlacZ lines produced spontaneous lung metastasis in nude mice. One of the lines, R3611-T2lacZ was highly efficient at metastatic conversion and produced more lung colonies than a faster growing v-raf/v-myc-transformed RJ2 14lacZ line. The spontaneously transformed RLElacZ line (C4T) was nonmetastatic, although it produced larger subcutaneous tumors than the metastatic R3611-T2lacZ line. Metastatic conversion correlated with upregulation of urokinase-type plasminogen activator receptor RNA expression and downregulation of plasminogen activator inhibitor-1, collagen alpha1 (I), and cytokeratin 14 (K14) RNA expression. These findings indicate that proteolytic activities associated with plasminogen activation play a role in the metastatic development in this model. Decreased production of extracellular proteins and cytoskeletal changes associated with lack of K14 expression are also likely to have contributed to the metastatic conversion of the RLE transformants. PMID- 9876220 TI - Tumor growth and metastasis of human colorectal cancer cell lines in SCID mice resemble clinical metastatic behaviors. AB - Ten human colorectal cancer (CRC) cell lines were implanted orthotopically into the ceca and also into the livers, muscles and peritoneal cavities of SCID mice in order to analyze the characteristics regulating metastatic behaviors of CRCs. All the CRC cell lines formed tumors in the muscle and cecum, but they could be classified into two groups: (1) six cell lines with high tumorigenicity in the liver (HTLs) forming differentiated tumors, and (2) four with no tumorigenicity in the liver (NTLs) forming poorly differentiated tumors in SCID mice. After orthotopic implantation, NTLs never metastasized to the liver, whereas HTLs did. Therefore, intrahepatic tumorigenicity and differential status were closely associated with liver metastasis whereas differentiation was not associated with lung metastasis. The 6 HTLs demonstrated an inverse correlation between liver metastases and peritoneal dissemination, and immunohistochemistry indicated expression of sLeX, CA19-9 and carcinoembryonic antigen in tumors which correlated well with the liver metastatic rate. We found a strong correlation between liver metastasis and intrahepatic tumorigenicity and could reproduce the clinical correlations between the pattern of the metastatic spread and the differentiation phenotype of CRC in vivo. We consider further examination using this model will be useful for analyzing the complex mechanisms involved in clinically metastasizing CRCs. PMID- 9876221 TI - Epidermal growth factor and laminin receptors contribute to migratory and invasive properties of gliomas. AB - Gliomas are characterized by their extensive invasion into the brain parenchyma. Recently it has been shown that normal brain cells can produce laminin, fibronectin and collagen type IV when confronted by invading glioma cells. Laminin stimulates cell migration of several human glioma cell lines in vitro. This migration can be inhibited by adding blocking monoclonal antibodies (MAbs) against the most expressed integrin subunits, alpha3 and beta1. Previous studies have shown that glioma cell migration, invasion and growth are stimulated by epidermal growth factor (EGF). However, MAb directed against the EGF receptor (EGFR) did only partly inhibit the invasive process in vitro. Since laminin has regional peptide homology with EGF (EGF-like repeats), the present work was aimed at studying how two human glioma cell lines exposed to antibodies to the EGFR, reacted to laminin stimulated migration. Furthermore, we wanted to study which role the EGFR and the laminin receptor integrin subunits alpha3 and beta1 play during glioma cell invasion. EGFR expression of two glioma cell lines, AN1/lacZ and U-251/lacZ was studied by flow cytometry and immunofluorescence microscopy. A cell migration assay was used to study effects of MAbs against EGFR on migration from laminin-stimulated tumor spheroids. Tumor cell invasion was evaluated by using an in vitro co-culture model, where normal fetal brain cell aggregates were confronted with multicellular tumor spheroids. The results show that both cell lines expressed EGFR, AN1/lacZ 4-fold more than U-251/lacZ. MAb against EGFR inhibited the laminin-stimulated migration only from AN1/lacZ spheroids. MAbs against alpha3 and beta1 integrin subunits inhibited glioma cell invasion in vitro. The present work indicates possible connections between laminin-stimulated cell migration and the EGFR expression on glioma cells. These elements contribute to the characteristic features of glioma cells and may be an important part of the complex relationships between growth factors, integrins and extracellular matrix during glioma cell invasion. PMID- 9876222 TI - The biological function of allergens: relevant for the induction of allergic diseases? AB - During the last 10 years an increasing number of allergens were identified and biochemically as well as immunologically characterized. More and more information on the primary structure of these molecules, revealed by recombinant technologies, became available. Furthermore for several allergens the biological functions could be elucidated. Most recently it was shown that beside IgE-binding capacity functional properties such as enzyme activity may contribute to the allergenicity and the induction of allergic inflammation. These findings could stimulate novel concepts in allergy prevention and therapy. This review summarizes recent data in this area. PMID- 9876223 TI - A species-specific monoclonal antibody to Cynodon dactylon. AB - BACKGROUND: It is usually difficult to differentiate between the pollens of different grass species on the basis of their appearance under a microscope, as they often appear similar. Such distinctions are important when interpreting the clinical relevance of pollens in air samples as individuals can differ in their allergic responses to different grass species. As this allergenic distinction occurs at the level of presence and differences of epitopes on the allergens associated with different species, it could be anticipated that species-specific monoclonal antibodies could provide such distinctions between pollens. METHOD: Monoclonal antibodies raised against Cynodon dactylon were screened and characterised in ELISA assays and blotting, using a range of grass pollen extracts, to identify clones which were species specific. RESULTS: The most specific monoclonal raised to C. dactylon did not react at a level of greater than 1.2% to extracts of 10 other grass pollens in a direct ELISA assay and showed no detectable cross-reactivity in a particle blotting assay. CONCLUSION: It has been possible to produce a monoclonal antibody that is functionally species specific to C. dactylon. PMID- 9876224 TI - Antigen interaction and heat inactivation expose new epitopes on human IgE. AB - It is well established that heat-denatured IgE is no longer capable of binding to FcepsilonRI. We have found an antibody that interacts with heat-denatured IgE. Interestingly, this antibody can also be used to detect some serum IgE, but not IgE synthesized de novo in vitro. However, native IgE can be transformed into an IgE that is recognized by this antibody, if antigen is added. Our data indicate that physiological mechanisms exist that biologically inactivate IgE which might still be mistaken for 'functional' IgE by assays based on polyclonal antibodies. PMID- 9876225 TI - Two common polymorphisms in the coding part of the CD43 gene are not associated with atopy. AB - BACKGROUND: Recently linkage and association of the chromosomal region 16p12-11 with enhanced IgE responsiveness have been shown. The gene coding for CD43 (sialophorin) has been localized to this region. Sialophorin represents a major sialoglycoprotein on the surface of human lymphocytes, monocytes and granulocytes. It is supposed to play an important role in human mast cell, T- and B-cell regulation and activation and has been described in connection with immunodeficiency diseases such as the Wiskott-Aldrich syndrome. Therefore, it can be designated as a candidate gene for atopy. METHODS: Using SSCP analysis and direct genomic sequencing, polymorphisms in the CD43 gene have been looked for and their association with atopy has been tested in a population of 260 largely atopic children and young adults. RESULTS: Three common polymorphisms in the coding part of the CD43 gene were found. Two of them are leading to amino acid exchanges, one from argine to cysteine at amino acid position 337 of the mature gene product and one from leucine to phenylalanine at amino acid position 341. Subsequent association studies revealed no obvious influence of R337C or L341F on IgE regulation (p = 0.47 and 0.43), neither in a cognate nor in an uncognate fashion. CONCLUSION: We conclude that CD43 polymorphisms are unlikely to account for the observed linkage effect at 16p12-11. Whether the polymorphisms R337C and L341F adjacent to phosphorylation sites in the intracellular region of the protein alter the normal functioning of CD43 remains to be elucidated. PMID- 9876227 TI - Effect of IL-4, IFN-gamma and IL-12 on cytokine production from human CD45RA and CD45RO CD4 T cell precursors. AB - The aim of this study was to compare the effects of IL-4, IL-12 and IFN-gamma on the production of T helper-1 (Th1) and T helper-2 (Th2)-type cytokines from human peripheral blood 'naive' CD45RA and 'memory' CD45RO CD4 T cells. CD45RA or CD45RO CD4 T cells were cultured for 4 days with phorbol myristate acetate (PMA) and ionomycin and either IL-4, IFN-gamma or IL-12 and their ability to proliferate and secrete IFN-gamma and IL-4 determined. Purified CD45RO CD4 T cells stimulated with PMA and ionomycin secreted higher levels of IL-4 and IFN-gamma, as measured by ELISA, than CD45RA CD4 T cells which secreted little IL-4 or IFN-gamma. However, CD45RA and CD45RO CD4 T cells proliferated to the same extent and IL-4, IFN-gamma and IL-12 had no effect on this. IL-12 and IFN-gamma had no effect on the amount of IL-4 secreted by PMA and ionomycin-stimulated CD45RO CD4 T cells, but culture with IL-4 enhanced IL-4 production 7-fold. IL-12 increased the amount of IFN-gamma produced by CD45RO CD4 T cells 2- to 3-fold. Small amounts of IFN gamma production were induced in CD4 CD45RA T cells by IL-12 and IFN-gamma. These results indicate: (1) that CD45RA cells cannot make significant amounts of IL-4 under the conditions used, (2) that CD45RO cells can produce both Th1 and Th2 cytokines immediately upon restimulation, (3) that IL-12 favours Th1 cytokine production in both CD45RA and CD45RO CD4 T cells, and (4) that IFN-gamma favours IFN-gamma production in CD45RA but not CD45RO cells. PMID- 9876226 TI - Granulocyte/macrophage-colony stimulating factor in allergen-induced rhinitis: cellular localization, relation to tissue eosinophilia and influence of topical corticosteroid. AB - BACKGROUND: Allergen-induced late nasal responses are associated with recruitment of T lymphocytes and eosinophils, and preferential messenger RNA (mRNA) expression of 'TH2-type' cytokines. We previously showed that topical steroid inhibited the late response and associated tissue eosinophilia. In this study we tested the hypothesis that granulocyte/macrophage-colony stimulating factor (GM CSF) may contribute to late-responses and tissue eosinophilia and is inhibitable by topical corticosteroid. METHODS: Nasal biopsies were taken before and 24 h after nasal allergen provocation following 6 weeks of treatment with either a nasal corticosteroid spray (fluticasone propionate) or a matched placebo nasal spray twice daily. Cryostat sections were processed by immunohistochemistry and in situ hybridization to assess cytokine mRNA expression for GM-CSF. RESULTS: Increases in T lymphocytes and eosinophils were seen in the nasal mucosa after allergen challenge (p = 0.01) which were accompanied by a 5-fold increase in cells expressing mRNA for GM-CSF (p = 0.01). Double immunohistochemistry/in situ hybridization demonstrated that the majority of GM-CSF mRNA+ cells were co localized to CD68+ (40%), or T cells (40%) with a lesser contribution from eosinophils (<20%). Topical steroid treatment was accompanied by a decrease in both the CD3+ and major basic protein (MBP+) cells expressing GM-CSF mRNA (p = 0.01) with a corresponding proportionate increase in the % of macrophages expressing GM-CSF. CONCLUSIONS: The results indicate that after allergen provocation, eosinophils are recruited to the nasal mucosa and that, at least in part, this may be due to GM-CSF. Topical nasal corticosteroid inhibits late responses and the associated eosinophilia, possibly indirectly by decreasing GM CSF from T lymphocytes or reducing autocrine production of GM-CSF from eosinophils. PMID- 9876228 TI - Serum levels of soluble interleukin-2 receptor at acute asthma exacerbation: relationship with severity of exacerbation and bronchodilator response. AB - BACKGROUND: T lymphocytes play a central role in the regulation of airway inflammation in asthma, and T cell activation appears to be a characteristic feature of acute asthma. It is not clear, however, whether this is proportional to the severity of acute asthma and is directly related to airway inflammation relevant to airflow obstruction during acute asthma. It is presumed that the extent to which bronchoconstriction or inflammation contributes to airflow obstruction in acute asthma may determine responsiveness to bronchodilator therapy. METHODS: Fifty asthmatic children who visited the emergency room due to acute exacerbation were studied. Serum levels of soluble interleukin-2 receptor (sIL-2R), a marker of T cell activation, were measured at the time of acute exacerbation and of clinical remission. At acute exacerbation, FEV1 was assessed before and after the administration of aerosolized salbutamol. RESULTS: The mean (+/-SD) serum level of sIL-2R at acute exacerbation (854+/-248 U/ml) was significantly higher (p<0.01) than at clinical remission (676+/-211 U/ml). It correlated positively with the severity of exacerbation (r = 0.47, p<0.01), but showed no significant relationship with bronchodilator response (r = -0.17, p = 0.20). CONCLUSION: A higher serum level of sIL-2R at acute exacerbation was associated with more severe exacerbation but not with lower bronchodilator response. These findings suggest that in the context of acute asthma, T cell activation is proportional to disease activity, but its relationship to airway inflammation relevant to the genesis of airflow obstruction remains obscure. PMID- 9876229 TI - Flow-cytometric evaluation of oxidative burst in phagocytic cells of children with cystic fibrosis. AB - OBJECTIVES: The objective of this study was to assess the dye 2', 7' dichlorofluorescein (DCF) assay in screening for alterations in polymorphonuclear cell (PMN) and monocyte (MC) oxidative burst of cystic fibrosis (CF) patients. STUDY DESIGN: 56 CF patients aged between 2 and 20 years were investigated. Purified cells were stimulated with phorbolmyristate acetate (PMA) and zymosan (ZX). A range for DCF fluorescence for PMA- and ZX-stimulated and non-stimulated cells was established based on data from 60 healthy controls. RESULTS: PMNs showed both enhancement and impairment. A deficient oxidative burst was detected in a total of 14 CF patients caused by abnormally high mean fluorescence intensity (MFI) of resting cells. Enhanced oxidative burst was seen in 6 CF patients. CF patients responded differently to PMA or ZX stimulation. Pseudomonas aeruginosa colonization significantly enhanced (p<0.005) the MFI of resting PMNs. MCs of CF patients showed a significantly (p<0.05) enhanced oxidative burst after stimulation with PMA compared to healthy controls, but no differences could be observed after stimulation with ZX. Serum concentrations of interleukin-6 were elevated in all CF patients, in particular in those with activation of both PMNs and MCs. CONCLUSION: The DCF assay shows for the first time the heterogeneity of the oxidative burst reaction in CF patients. In our opinion, the DCF assay is a reliable method for detecting pathological oxidative burst in CF patients. PMID- 9876230 TI - Use of cultured thymic tissues for the regeneration of the thymus. AB - An account of research conducted on the transplantation of thymic cells and tissues in order to restore the functional activity of the thymus is reviewed, and discussed in the context of current concepts. Although most rodent work has been conducted on the transplantation of cultured fragments under the kidney capsule, human transplantation studies and models have used other sites or other species such as the severe combined immunodeficient mouse as hosts. The factors affecting the growth of thymic cells in culture is considered in detail since the methodology can strongly influence the cells favoured under culture conditions. An extension of this work to characterize both thymic fragments implanted under the kidney capsule of rats and cultured thymic cells has recently led to the appreciation that the adult thymus must contain a small number of neural crest like cells. These cells have a high level of proliferation in the implanted fragments, expand in culture, and belong to a family of cytokeratin-positive cells exhibiting immunoreactivity for a wide range of neuropeptides and transmitters. Thus primary cultures of thymus can contain a wide range of glia like cells. This can be explained by the fact that the thymus, in addition to having a mesenchymal neural crest component, is probably derived from cardiac neural crest. Closely associated neural crest also has glia-like properties (the supporting cells of the enteric nervous system). These finding can account for the large number of reports of epithelial cells of the adult thymus being immunoreactive to antibodies raised to neuroendocrine and neurotransmitters. Neuroimmune interactions in the thymus are more fundamental than previous work had suggested. PMID- 9876231 TI - Accelerated maturation of the thymic stroma in the progeny of adrenalectomized pregnant rats. AB - The possible role played by glucocorticoids (GCs) in the development of thymic stromal cell components has been studied in the progeny of adrenalectomized pregnant rats (FAdx), an experimental model which ensures the absence of GCs until the establishment of the fetal hypothalamus-pituitary gland-adrenal gland axis. As previously demonstrated for thymocytes, the lack of GCs early in ontogeny results in an accelerated maturation of the thymic stromal elements. Early expression of specific cell markers for thymic epithelial cell subsets and appearance of a well-established cytokeratin-positive epithelial cytoreticulum confirmed the ultrastructural evidence of a faster maturation of the thymic epithelium in FAdx than in FSham. A similar faster and stronger pattern of both class I and class II molecule expression on the epithelial cells occurred in the former fetuses than in control ones. Changes in the pattern of expression of laminin, but not that of fibronectin, throughout thymic maturation also reflected accelerated maturation. Immunohistochemically identified thymic macrophages appeared late in both FSham and FAdx but in higher numbers in these latter indirectly demonstrating their faster development. Finally, the maturation and turnover of thymic dendritic cells showed a remarkable acceleration in the FAdx. In 15- to 16-day-old FAdx thymuses there was a high number of dendritic cells which sharply decreased in the following days suggesting a massive migration to the periphery and/or in situ cell death. In parallel a new wave of dendritic cell progenitors began to differentiate in the FAdx thymuses but not in the FSham ones. The results are discussed from the view of close relationships known to occur between thymocytes and the stromal components, although a direct effect of GCs cannot be discarded. PMID- 9876232 TI - Rapid progesterone actions on thymulin-secreting epithelial cells cultured from rat thymus. AB - Many soluble factors of neural, endocrine, paracrine and autocrine origin are present in the thymus and modulate its function. Long-term effects of sex steroids have been documented for thymocytes and cells of the thymic microenvironment. In this report we examine rapid actions of progesterone upon aspects of epithelial cell physiology. Progesterone (0.1-10 microM) was applied to cultured thymulin-secreting thymic epithelial cells (TS-TEC) and changes in transmembrane potential, transmembrane current, intracellular calcium levels and thymulin secretion were assessed. Rapid changes in electrophysiology and intracellular calcium provide evidence for a membrane-bound progesterone receptor in these cells, in addition to classical cytoplasmic receptors. Application of progesterone to TS-TEC caused electrophysiological changes in 56% of cells (n = 40), activating an inward current (-24 +/- 9 pA at 1 microM, n = 7, p < 0.02) and dose-dependent depolarization (7.1 +/- 1.8 mV at 1 microM, n = 19, p < 0.01). Intracellular calcium levels, monitored by the ratiometric fluorescent calcium indicator fura-2, increased within seconds of progesterone (1 microM) application. Progesterone (1 microM) increased thymulin levels in supernatant, as measured by ELISA, above the levels in the preapplication period (142 +/- 16% of the preapplication period, n = 3, p < 0.02). This effect was reduced in the presence of cobalt chloride which blocks voltage-dependent calcium channels. In addition, TS-TEC in culture were immunoreactive to antibody AG7. This antibody was raised to a membrane-bound antigen involved in calcium influx subsequent to progesterone binding in sperm. Thus we suggest that progesterone acts upon many aspects of TS-TEC physiology through both cytoplasmic and membrane-bound receptors. PMID- 9876233 TI - Cytokine-mediated or direct effects of thymulin on the nervous system as assessed by pain-related behavior. AB - Thymulin is a thymic hormone with known immunomodulatory activities. Recent evidence has indicated a signaling role for this peptide in the interaction between the immune, endocrine and the nervous system. In this report, we review recent experimental findings on the analgesic actions of thymulin (high doses) in rats with endotoxin-induced localized inflammation and the hyperalgesic actions (low doses) of this peptide in intact animals. These actions involve both proinflammatory cytokines and PGE2. The possibility of a dual role played by thymulin as a hormone that might also involve a direct effect on the nervous system is discussed. PMID- 9876234 TI - Effect of transmitters and co-transmitters of the sympathetic nervous system on interleukin-6 synthesis in thymic epithelial cells. AB - In the thymus, sympathetic nerves run in septa in close connection to subcapsular/perivascular thymic epithelial cells (TEC). Since TEC are supposed to create a microenvironment of cytokines necessary for the development of thymocytes to T cells, we investigated the influence of sympathetic transmitters and co-transmitters on interleukin-6 (IL-6) synthesis in cultivated rat TEC that express markers of perivascular/subcapsular TEC. Noradrenaline and ATP stimulated IL-6 production in the culture supernatants 14- and 23-fold over basal values after 24 h. Co-stimulation with noradrenaline and ATP yielded an additive effect. Synthesis of IL-6 was concentration-dependent upon ATP and appeared to be mediated by P2 purinoceptors. During 24 h stimulation with 1 mM ATP, two thirds of the ligand was degraded mainly to ADP, production of AMP and adenosine was minor or negligible. Thus, in TEC, transmitters and co-transmitters of the sympathetic nervous system have a co-stimulatory effect on synthesis of IL-6 that is an important factor for thymocyte differentiation and proliferation. PMID- 9876235 TI - A possible role for acetylcholine in the dialogue between thymocytes and thymic stroma. AB - In this article we will review data suggesting that acetylcholine takes part in the mutual interplay between developing T cells and thymic epithelium, and thereby may influence the generation of the T-cell repertoire. In the first part we will recapitulate our findings according to which cholinergic agonists affect thymocyte apoptosis via a nicotinergic effect on thymic epithelial cells. In the second part we will present evidence that acetylcholine within the thymus is mainly derived from the thymocytes themselves, and that the production and release of this neurotransmitter is dependent on activation of thymic lymphocytes. PMID- 9876236 TI - The immune-endocrine loop during aging: role of growth hormone and insulin-like growth factor-I. AB - Why a primary lymphoid organ such as the thymus involutes during aging remains a fundamental question in immunology. Aging is associated with a decrease in plasma growth hormone (somatotropin) and IGF-I, and this somatopause of aging suggests a connection between the neuroendocrine and immune systems. Several investigators have demonstrated that treatment with either growth hormone or IGF-I restores architecture of the involuted thymus gland by reversing the loss of immature cortical thymocytes and preventing the decline in thymulin synthesis that occurs in old or GH-deficient animals and humans. The proliferation, differentiation and functions of other components of the immune system, including T and B cells, macrophages and neutrophils, also demonstrate age-associated decrements that can be restored by IGF-I. Knowledge of the mechanism by which cytokines and hormones influence hematopoietic cells is critical to improving the health of aged individuals. Our laboratory has recently demonstrated that IGF-I prevents apoptosis in promyeloid cells, which subsequently permits these cells to differentiate into neutrophils. We also demonstrated that IL-4 acts much like IGF I to promote survival of promyeloid cells and to activate the enzyme phosphatidylinositol 3'-kinase (PI 3-kinase). However, the receptors for IGF-I and IL-4 are completely different, with the intracellular beta chains of the IGF receptor possessing intrinsic tyrosine kinase activity and the alpha and gammac subunit of the heterodimeric IL-4 receptor utilizing the Janus kinase family of nonreceptor protein kinases to tyrosine phosphorylate downstream targets. Both receptors share many of the components of the PI 3-kinase signal transduction pathway, converging at the level of insulin receptor substrate-1 or insulin receptor subtrate-2 (formally known as 4PS, or IL-4 Phosphorylated Substrate). Our investigations with IGF-I and IL-4 suggest that PI 3-kinase inhibits apoptosis by maintaining high levels of the anti-apoptotic protein Bcl-2. The sharing of common activation molecules, despite vastly different protein structures of their receptors, forms a molecular explanation for the possibility of cross talk between IL-4 and IGF-I in regulating many of the events associated with hematopoietic differentiation, proliferation and survival. PMID- 9876237 TI - Use of neuroendocrine hormones to promote reconstitution after bone marrow transplantation. AB - A survey of the previous literature and the data shown here indicate that neuroendocrine hormones such as growth hormone and prolactin may be of potential clinical use after bone marrow transplantation (BMT) to promote hematopoietic and immune recovery. The amounts of hormones used in our model do not promote weight gain suggesting that their lymphohematopoietic actions were independent of their anabolic effects. While the hormones may not produce the same extent of immune/hematopoietic effects when compared to conventional hematopoietic and immune stimulating cytokines (i.e. IL-2 or G-CSF), their pleiotropic effects and limited toxicity after systemic administration makes them attractive to test in the post-BMT setting. However, more work needs to be performed to understand the mechanism(s) of their action, particularly with regard to T-cell function and development. PMID- 9876238 TI - Extracellular ATP: a further modulator in neuroendocrine control of the thymus. AB - It is well established that the process of thymocyte differentiation and maturation occurs in the thymus, where cell-to-cell communication is essential for providing the messages to T-cell precursors. At least two pathways are important for such communication: one via membrane surface molecules and the other via soluble mediators such as cytokines and some hormones. Recently, the presence of receptors for extracellular ATP has been demonstrated on thymocytes and microenvironment cells, and putative functions for this molecule have been proposed. Herein we focus on the recent evidence which supports the view of extracellular ATP and some related nucleotides as novel intrathymic signal molecules. In addition, we discuss the possible physiological implications of such purinergic receptors for the physiology of the thymus. PMID- 9876239 TI - Thymus-derived glucocorticoids and the regulation of antigen-specific T-cell development. AB - Bidirectional interactions of both a stimulatory and inhibitory nature occur between the neuroendocrine and the immune systems, and these interactions play an important modulatory role during T-cell ontogeny. Specifically, glucocorticoids potently induce apoptosis in thymocytes and activated T cells, but can also rescue these cells from activation-induced cell death. The objective of this review is to discuss current data on the interactions of the immune system with steroid hormones in the thymus and to describe a model that includes glucocorticoids in the shaping of the peripheral T-cell antigen-specific repertoire and deals with their potential role in the generation of autoimmune disease. PMID- 9876240 TI - Vasoactive intestinal peptide in thymus: synthesis, receptors and biological actions. AB - Evidence summarized in this report indicates that thymocytes produce and secrete VIP. Moreover, different stimuli such as Con A, LPS and anti-TCR antibody induce a significant increase in VIP production by thymocytes. In addition, proinflammatory cytokines such as IL-1, IL-6 and TNF-alpha, but not IL-2, stimulate in a similar time-dependent manner VIP production by lymphocytes. We also describe the expression of VIP1 receptor and VIP2 receptor mRNA in murine thymocytes. Thus, VIP released in thymus microenvironment may modulate immune functions through direct binding to VIP receptors on thymocytes. Our functional data support that VIP through the interaction with their specific receptors affect three important aspects of thymocytes function: cytokine production, mobility and apoptosis. PMID- 9876241 TI - Thymic expression of the pancreatic endocrine hormones. AB - The thymus plays a central role in the selection of T lymphocytes that are tolerant to 'self' antigens and responsive to foreign pathogens. We and others have reported the expression of the pancreatic endocrine hormones, preproinsulin, proglucagon, prosomatostatin and propancreatic polypeptide in the human and mouse thymus. While mRNA expression is very low there is evidence for the presence of the translated product. In addition, we have investigated the cell types responsible for expression. In the thymus, hormone expression is enriched in the antigen-presenting cell population. Interestingly, while proglucagon, prosomatostatin and propancreatic polypeptide appear to be expressed in a macrophage population, preproinsulin expression was restricted to dendritic cells which are more potent antigen-presenting cells. The functional significance of the endogenous expression of insulin in the thymus has been indirectly investigated using transgenic models in which the transgene is introduced by the rat insulin promoter. The data suggest that thymic expression of the transgene is critical in the induction of T-cell tolerance to the transgene in the periphery. Taken together, the evidence suggests that the low-level pancreatic hormone expression in the thymus may be involved in central tolerance to proteins of restricted expression. PMID- 9876242 TI - The thymic repertoire of neuroendocrine-related self antigens: biological role in T-cell selection and pharmacological implications. AB - Thymic epithelium, including nurse cells (TEC/TNC), as well as other thymic stromal cells (macrophages and dentritic cells), express a repertoire of polypeptide belonging to various neuroendocrine protein families (such as the neurophypophysial, tachykinin, neurotensin and insulin families). A hierarchy of dominance exists in the organization of the thymic repertoire of neuroendocrine precursors. Oxytocin (OT) is more expressed in the TEC/TNC than vasopressin (VP); insulin-like growth factor 2 (IGF-2) thymic expression predominates over IGF-1, and much more over (pro)insulin. Thus, OT was proposed to be the self antigen of the neurohypophysial family, and IGF-2 the self antigen precursor of the insulin family. The dual role of the thymus in T-cell life and death is recapitulated at the level of the thymic neuroendocrine protein repertoire. Indeed, thymic polypeptides behave as accessory signals involved in T-cell development and positive selection according to the cryptocrine model of signaling. Moreover, thymic neuroendocrine polypeptides are the source of self antigens presented by thymic MHC molecules to developing pre-T cells. This presentation might induce the negative selection of T cells bearing a randomly rearranged antigen receptor (TCR) oriented against neuroendocrine families. Using an animal model of autoimmune type 1 diabetes (BB rat), we have shown a defect in intrathymic expression of the self antigen of the insulin family (IGF-2) and in IGF-2 mediated T-cell education to recognize and tolerate the insulin family. Altogether these studies have enlightened the crucial role played by the thymus in the induction of the central self tolerance of neuroendocrine families. The tolerogenic properties of thymic self peptides could be used in a novel type of vaccination for the prevention of autoimmune diseases. PMID- 9876243 TI - Immunoneuroendocrine connectivity: the paradigm of the thymus hypothalamus/pituitary axis. AB - It is now largely established that the immune and neuroendocrine systems cross talk by using similar ligands and receptors. In this context, the thymus hypothalamus/pituitary axis can be regarded as a paradigm of connectivity in both normal and pathological conditions. For example, cytokines and thymic hormones modulate hypothalamic-pituitary functions: (a) interleukin (IL)-1 seems to upregulate the production of corticotropin-releasing factor and by adrenocorticotropin by hypothalamic neurons and pituitary cells, respectively; (b) thymulin enhances LH secretion. Conversely, a great deal of data strongly indicate that the hypothalamic-pituitary axis plays a role in the control of thymus physiology. Growth hormone (GH) for example, enhances thymulin secretion by thymic epithelial cells (TEC), both in vivo and in vitro, also increasing extracellular matrix-mediated TEC/thymocyte interactions. Additionally, gap junction-mediated cell coupling among TEC is upregulated by ACTH. In a second vein, it was shown that GH injections in aging mice increased total thymocyte numbers and the percentage of CD3-bearing cells, as well concanavalin-A mitogenic response and IL-6 production. In addition to mutual effects, thymus-pituitary similarities for cytokine and hormone production have been demonstrated. Cytokines such as IL-1, IL-2, IL-6, interferon-gamma, transforming growth factor beta and others can be produced by hypothalamic and/or pituitary cells. Conversely, hormones including GH, PRL, LH, oxytocin, vasopressin and somatostatin can be produced intrathymically. Moreover, receptors for various cytokines and hormones are expressed in both the thymus and the hypothalamus/pituitary axis. Lastly, it is noteworthy that a thymus-pituitary connectivity can also be seen under pathological situations. In this regard, an altered HPA axis has been reported in AIDS, human falciparum malaria and murine rabies, that also show a severe thymic atrophy. PMID- 9876244 TI - The thymus-pituitary axis and its changes during aging. AB - The pituitary-thymic axis constitutes a bidirectional circuit where the ascending feedback loop is effected by thymic factors of epithelial origin. The aim of the present article is to review the evidence demonstrating that aging brings about a progressive disruption in the integration of this network. In doing so, we briefly review the experimental evidence supporting the view that immune and neuroendocrine aging are interdependent processes. The advantages and limits of the nude mouse as a model of thymus-dependent accelerated aging is also discussed. Next, we review a number of studies which show that the endocrine thymus produces several bioactive molecules, generally called thymic hormones, which in addition to possessing immunoregulatory properties are also active on nervous and endocrine circuits. In particular, the reported activities of thymosin fraction 5 (TF5), thymosin alpha-1 and thymosin beta-4 on beta endorphin, ACTH, glucocorticoids, LHRH and LH secretion in different animal and cell models are reviewed. The known hypophysiotropic actions of other thymic hormones like thymulin, homeostatic thymus hormone (HTH) and thymus factor are summarized. Aging has a significant impact on pituitary responsiveness to thymic hormones. Thus, it has been reported that TF5 and HTH have thyrotropin-inhibiting activity in young but not in old rats. Furthermore, intravenous administration of HTH was also able to reduce plasma GH and increase corticosterone levels in both young and old rats, although these responses were much weaker in the old animals. Further evidence on this topic is discussed. It is proposed that in addition to its central role in the regulation of the immune function, the thymus gland may extend its influence to nonimmunologic components of the body, including the neuroendocrine system. The early onset of thymus involution might therefore act as a triggering event which would initiate the gradual decline in homeostatic potential that characterizes the aging process. PMID- 9876245 TI - Primitive cerebral neuroectodermal tumors excluding medulloblastomas: a retrospective study of 30 cases. AB - We present a retrospective study of 30 cases of primitive cerebral neuroectodermal tumors (PNET), excluding medulloblastomas, referred to us postoperatively for additional therapy to evaluate prognostic factors and treatment efficiency. The histologic types were: pinealoblastomas (n = 7); ependymoblastomas (n = 2); medulloepitheliomas (n = 4), and other PNET (n = 17). The tumor was located in the supratentorial area in 24 patients and in the posterior fossa in 6 patients. Among the supratentorial tumors, 8 were metastatic. Maximal surgical resection was performed. Sixteen of 30 patients had no measurable disease after surgery and were considered as standard-risk (SR) cases, and 14 with a local residue or metastasis as high-risk (HR) cases. The objective of postsurgical treatment was to avoid radiotherapy in children below 4 years of age. It consisted of radiotherapy alone in 6 patients, chemotherapy alone in 17, and radiotherapy with chemotherapy in 7. Furthermore, high-dose chemotherapy (busulfan, thiotepa) and autologous bone marrow transplantation, performed in 6 patients, yielded a response rate of 3/6. Event-free survival (EFS) of SR patients was 37% at 3 years (95% confidence interval (CI) 14-60%) and overall survival 44% (95% CI 26-62%). Only 1 of the HR patients achieved a complete remission and all of them died early. The critical prognostic factors appear to be the completeness of initial surgical resection and absence of metastasis. These tumors have a poor prognosis. Novel strategies (high-dose chemotherapy) are needed to improve their outcome because the children concerned are very young and the effects of radiotherapy are particularly deleterious when tumors are situated in the supratentorial area. PMID- 9876246 TI - Intramedullary spinal cord ependymomas in children: treatment, results and follow up. AB - A retrospective review of 20 pediatric patients with intramedullary spinal cord ependymomas, all of whom underwent operative resection between 1985 and 1996, was undertaken to determine surgical results, long-term follow-up and tumor recurrence. Twelve children operated on in the same period with filum or cauda equina ependymomas were not included in this study. Nine children had had previous treatment before referral. Gross total resection was achieved in 14 patients and subtotal in 6. None of these had a post-operative radiation therapy. The median follow-up period was 67 months (range 25-177 months). All children were clinically evaluated before and after operation and at the last follow-up. The clinical grade at the last follow-up showed improvement in 8 patients (40%), was unchanged in 10 (50%) and deteriorated in 2 (10%). Three patients had a recurrence, 2 at the primary site (2 and 3 years after our surgery) and 1 at a distant site (3 years after). The actuarial 5- and 10-year survival rates were both 90%; 5- and 10-year progression-free survival rates were 93 and 70%, respectively. We conclude that a complete removal can be achieved in almost all cases of intramedullary spinal cord ependymomas in children, and that the long survival rates justify avoiding post-operative radiation therapy. PMID- 9876247 TI - Recurrent tethering: a common long-term problem after lipomyelomeningocele repair. AB - The authors reviewed the records of 94 patients who underwent initial repair of a lipomyelomeningocele between 1982 and July 1996 at the Children's Hospital of Pittsburgh to determine the incidence and time course of symptomatic retethering. In each of these patients, the initial operative goals were to microsurgically debulk as much of the lipoma as possible to allow the conus to move freely within the spinal canal, to divide any tethering arachnoidal adhesions, to close the pia if possible and to reconstitute a capacious thecal sac, using a dural graft if necessary. With a median follow-up of 58 months, 19 of these patients (20.2%) required 28 subsequent operations for symptomatic retethering. Median time between the initial procedure and reoperation for retethering was 52 months. The primary complaint of 12 patients was intractable low-back or leg pain. Other common symptoms were progressive bowel and/or bladder dysfunction, deterioration of motor function and foot deformities. The decision to reoperate was based predominantly on the clinical situation of the patient; magnetic resonance imaging was used to confirm the location and extent of tethering. Patients with transitional lipomas had a significantly higher frequency of symptomatic retethering than those with caudal or dorsal lesions (p < 0.05). No other clinical or technical feature correlated with an increased frequency of retethering. In particular, none of a variety of types of dural graft materials appeared to entirely prevent symptomatic retethering. Following reoperation, pain complaints resolved and many of the other symptoms improved partially or resolved completely. Although the long-term results were also favorable in the majority of patients, a small subgroup (n = 6) exhibited repetitive symptomatic tethering that proved increasingly difficult to treat. We concluded that symptomatic retethering is a common problem in children with lipomyelomeningoceles, even after an adequate initial operation. To date, no type of graft material has been shown to entirely prevent this problem. Close long-term surveillance of such patients is required to allow detection and treatment of symptomatic retethering. PMID- 9876248 TI - Upper extremity performance and self-care skill changes in children with spastic cerebral palsy following selective posterior rhizotomy. AB - Changes in upper extremity and self-care performance following selective posterior rhizotomy (SPR) are reported frequently, but rarely quantified. In this study, 36 children with spastic cerebral palsy were assessed preoperatively and 1 year following SPR using the Quality of Upper Extremity Skills Test (QUEST). Twenty-six children were assessed at similar intervals using the Functional Independence Measure for Children (WeeFIM) as a measure of self-care performance. Wilcoxon matched-pair signed-rank tests were used to compare the QUEST total scores and the WeeFIM motor, cognitive, and total scores before and after SPR. One year after SPR, the total QUEST scores were significantly better (median improvement = 3.2%, p < 0.0001), as were the WeeFIM motor (median improvement = 9.5, p < 0. 0001), cognitive (median improvement = 1.0, p < 0.008), and total (median improvement = 11.0, p < 0.0001) scores. The results indicate quantifiable improvements in upper extremity function, and motor and cognitive self-care skills in children 1 year after SPR. PMID- 9876249 TI - Intracranial germinoma associated with Down's syndrome. Report of 2 cases. AB - It is well known that Down's syndrome is sometimes associated with leukemia. However, there have been only a few case reports of a relationship between Down's syndrome and brain tumors. We report 2 cases with histological diagnoses of germinoma. The 1st case was a 10-year-old boy with Down's syndrome complaining of seizure and left hemiparesis. Computed tomographic (CT) scan and magnetic resonance imaging (MRI) showed a mass lesion in the right basal ganglia and thalamus. Histological examination indicated two cell pattern germinomas. The 2nd case was a 20-year-old man with Down's syndrome complaining of headache and vomiting. CT scan and MRI showed a pineal region tumor with marked hydrocephalus. Surgical specimens showed typical germinoma. Only 13 cases of brain tumors associated with Down's syndrome have been reported. A higher incidence of germ cell tumors seems to be related to chromosomal abnormalities. PMID- 9876250 TI - Supratentorial dorsal cistern epidermoid cyst in childhood. AB - Epidermoid cysts (ECs) are rare developmental lesions occurring anywhere along the central nervous system. Usually affecting adult patients, the occurrence of supratentorial dorsal cistern ECs has been reported in only 1 certain case of pediatric age. An additional pediatric case is presented. An 11-year-old boy had an 8-year history of petit mal seizures. Neurologic examination on admission was negative. T1-weighted magnetic resonance images demonstrated homogenous, hypointense, intradural, extracerebral, right frontal, precentral, parasagittal mass with relatively high signal intensity on T2-weighted images. The lesion was completely removed using a microneurosurgical technique with special care taken with regard to the adjacent nervous and vascular structures. Histologically, an EC was diagnosed. The patient had no recurrence for 2 years and seizures were significantly reduced. This case suggests the need to include ECs in the differential diagnosis of intradural supratentorial extracerebral lesions in childhood. PMID- 9876252 TI - Images in pediatric neurosurgery. Neuroimaging quiz. PMID- 9876251 TI - Familial intracranial arteriovenous malformations. Case report and review of the literature. AB - Familial intracranial arteriovenous malformations (AVMs) are rare. We describe a father and son both presenting in childhood with cerebral AVMs. Both patients underwent successful surgical extirpation of the AVM. Familial AVMs present at a young age and tend to be multiple, as was the case in one of our patients. Imaging of asymptomatic family members may detect previously undiagnosed AVMs, and we recommend screening of the relatives of patients with a strong family history of such lesions. PMID- 9876253 TI - Fusiform P1 segment artery aneurysm in a pediatric patient: technical case report. AB - We present an unusual aneurysm in a pediatric patient. Due to the fusiform nature of the aneurysm and the small size of the patient, a unique surgical solution was applied. One year of clinical follow-up is also provided. PMID- 9876254 TI - Concerning the article by Andar UB et al. Pediatr Neurosurg 1997; 26: 17-24: Upper and lower motor neuron dysfunction in closed spinal dysraphism. PMID- 9876255 TI - Maintenance of hippocampal cell numbers in young and aged rats submitted to chronic unpredictable stress. Comparison with the effects of corticosterone treatment. AB - Exposure of rats to sustained stress has been associated with behavioural impairments, the degree of impairment being greater with increasing age of the subject. Although the behavioural deficits have been frequently attributed to stress-induced neuronal loss in the hippocampus, the validity of that view may be disputed since it is based on data collected using conventional morphometric methods which are subject to bias. The question of whether stress per se does indeed induce hippocampal cell losses was therefore re-examined using unbiased stereological tools in the present work. Specifically, we used the optical fractionator and the Cavalieri principle, to respectively estimate the total number of neurons and volumes of the main divisions of the hippocampal formation of young and old rats which had been exposed for 1 month to an unpredictable stress paradigm. The efficacy of the treatment was confirmed by elevated serum corticosterone levels measured at various intervals during the experimental period. In order to evaluate whether any deleterious effects might have occurred merely due to the stress-induced elevations in corticosterone secretion, we conducted a parallel study on animals that were injected with corticosterone over a similar duration. Neither stress nor treatment with corticosterone was found to result in significant cell losses in any division of the hippocampal formation; likewise, neither treatment produced significant volumetric differences. Further, these results were not influenced by age of the experimental subjects. The present findings therefore call for a reappraisal of the hypothesis that hippocampal cell loss accounts for the behavioural impairments observed by others following prolonged stress and/or chronic elevation of serum corticosterone levels. PMID- 9876256 TI - The effect of repeated exposure to forced swimming on extracellular levels of 5 hydroxytryptamine in the rat. AB - The effects of repeated exposure to forced swimming was examined on extracellular concentrations of 5-hydroxytryptamine (5-HT), behavioral, and endocrine responses in rats. Animals were exposed to a 15-min swimming session on two consecutive days. On the first day, the swimming session increased extracellular 5-HT by 80 % over baseline in the striatum and reduced 5-HT to 40 % below baseline in the lateral septum. On the second day, however, the swimming session produced no effect on 5-HT in either brain region. Ratings of behavior showed that rats demonstrated climbing and swimming behaviors and developed immobility during the initial swimming session and that ratings of immobility increased and swimming decreased during the second swimming session. Immobility was positively correlated and swimming was negatively correlated with changes in extracellular 5 HT in the lateral septum but not in the striatum. Plasma corticosterone was equally elevated by 950 % after either 1 or 2 days of swimming exposure. These results show that there is rapid adaptation to the effects of repeated forced swimming on the regionally-specific, bi-directional response of extracellular 5 HT. In addition, changes in extracellular 5-HT in the lateral septum may be related to the behaviors produced during the forced swimming test that underlie its utility as an animal model of depression. PMID- 9876257 TI - Increased heat shock protein expression after stress in Japanese quail. AB - Heat shock proteins (HSPs) have been shown to provide information on the biological impact of environmental stress to organisms, yet none have investigated the HSP response to stress in birds. Japanese quail were exposed to seven different stressors (mild restraint, loud noise, inescapable irritation, cold temperature, isolation in darkness, and two stressful social situations) and expression of HSP30, 60, 70, and 90 in heart, liver, lung, kidney and gonads was examined. Tonic Immobility (TI) tests were also conducted to assess whether the stressors increased fear response. Increased expression of HSP70 was found in the myocardial tissue of birds exposed to loud noise, inescapable irritation, cold temperature, and isolation in darkness. Increased expression of other HSPs was not apparent in the heart or any of the other all tissues examined. Longer TI was observed only in birds exposed to the noise stress. Evidence is presented that a fairly wide range of stressors caused increased expression of HSP70 in the Japanese quail myocardial tissue and that HSPs may provide useful biomarkers for the study of environmental stress in birds. PMID- 9876258 TI - Chronic prenatal stress affects development and behavioral depression in rats. AB - We have used the approach of Willner et al (1987), which consists of transitory and variable changes in the rat"s living conditions, to investigate the influence of chronic prenatal stress on pup development and their susceptibility to behavioral depression at adult age, as assessed by the learned helplessness model. Pregnant female Wistar rats were divided into either stressed (S; N = 35) or non-stressed (NS; N = 35) groups during the last two weeks of pregnancy. The male and female pups of both groups were either handled to test for physical development up to weaning (H; N = 25 litters) or left undisturbed (NH; N = 10 litters) until adult age, at which time the males from all four experimental groups were divided into two subgroups (N = 10 each) and were submitted to the learned helplessness model of depression. Prenatal stress reduced the number of male pups per litter, decreased the anogenital distance, and produced earlier earflap and eye opening dates, as well as a faster righting. Behavioral depression was induced in all cases, except in the NS-H animals. The prenatally stressed, non-handled pups showed greater escape latency than the NS subgroups. We conclude that the stress schedule used in this study was stressful to dams and sufficient to affect the pups" development and to increase the intensity of induced behavioral depression at adult age. PMID- 9876259 TI - Effects of major depression, aging and gender upon calculated diurnal free plasma cortisol concentrations: a re-evaluation study. AB - Depression, aging and female gender are associated with increased diurnal concentrations of total plasma cortisol. For the physical effects of hypercortisolemia, however, it is generally assumed that free rather than total plasma cortisol concentrations are of importance. Herein, we report a mathematical approach to determine free plasma cortisol concentrations on the basis of total cortisol, corticosteroid binding-globulin (CBG) and albumin plasma concentrations. This approach was used to re-evaluate two sets of data in order to estimate the effect of depression as well as the effect of aging and gender upon free plasma cortisol concentrations. Comparing male depressed patients with healthy controls, we found 24-hour free cortisol minima (MIN: 4.1 +/- 1.8 vs. 1.6 +/- 1.1 nmol/l, p < 0.0001), mean (MEAN: 25.5 +/- 6.7 vs. 10.4 +/- 2.7 nmol/l, p < 0.0001) and maximal (MAX: 85.3 +/- 23.3 vs. 45.2 +/- 15. 8 nmol/l, p < 0.0001) concentrations to be significantly increased in depressed patients. In general, the impact of depression upon total plasma cortisol were not only maintained, but stronger regarding free plasma cortisol. Also, age was associated with free plasma cortisol MIN (F1,30= 10.8, p < 0.003) and free plasma cortisol MEAN (F1,30 = 8.9, p < 0.006). All effects of age upon total plasma cortisol were generally also found in free plasma cortisol, though with less impact. No effect of gender upon any of the given free plasma cortisol outcome variables was found. Taken together, our re-evaluation clearly shows not only depression but also aging to be associated with increases in free plasma cortisol concentrations. This finding is in line with the observation that in both conditions medical problems triggered and/or maintained by glucocorticoids (e.g. osteoporosis) are frequently seen. PMID- 9876260 TI - Selective in vivo stimulation of stress-activated protein kinase in different rat tissues by immobilization stress. AB - Stress activated protein kinases (SAPK) are key enzymes mediating the cellular response to stressful stimuli. While they are intensively studied in cultured cells, little is known about their physiological role in vivo, or relevance to pathological conditions. Therefore we examined the effect of various times of immobilization on c-Jun N-terminal protein kinase (JNK) activity in several rat stress responsive tissues and in a number of other locations. The abundance and relative distribution of JNK isoforms, the basal levels, time course and relative magnitude of stress induced JNK activity differed among tissues and regions of the brain of the same animal. JNK immunoreactive proteins were most abundant in the brain, especially in the hippocampus, hypothalamus and frontal cortex. Marked activation in response to immobilization stress was observed in adrenal medulla, adrenal cortex, aorta and hippocampus, less pronounced in locus coeruleus. JNK was not affected in superior cervical ganglia, pituitary, hypothalamus, frontal cortex and cerebellum. In adrenal medulla, the activation of JNK by single immobilization stress is correlated with increased transcription of stress responsive genes, tyrosine hydroxylase and dopamine beta-hydroxylase. These data suggest a potential role of JNK signal transduction pathway in mediating the long term adaptation to stressful stimuli in vivo. PMID- 9876261 TI - Tropical paediatrics and international child health in Liverpool, 1968-1998. PMID- 9876262 TI - Experiences of work among refugees. PMID- 9876263 TI - HIV infection in children: the widening gap between developing and industrialized countries. PMID- 9876264 TI - AIDS hospices and orphanages. PMID- 9876265 TI - Community-based paediatric curriculum: the Malawi experience. PMID- 9876266 TI - Prevalence of childhood asthma in the tropics. AB - There is evidence that until quite recently asthma was infrequent in tropical developing countries. Estimates of the current and lifetime prevalence of wheeze and asthma in children in tropical countries in the 1990s suggest that the problem is increasing. There is also evidence that urbanization in tropical areas is associated with a higher prevalence of exercise-induced bronchospasm. Possible contributors to this changing pattern of asthma are discussed. These probably relate to differential exposure to house dust mite and infectious agents in infancy. There is little research on this increasing problem but it is needed in order to find ways to arrest increases in asthma. PMID- 9876267 TI - Investigations into the presence of aflatoxins in human body fluids and tissues in relation to child health in the tropics. PMID- 9876268 TI - Fumonisins and human health. AB - Fumonisins are mycotoxins produced by Fusarium moniliforme that are prevalent in corn, sorghum, millet and other agricultural products. It is possible that fumonisins are aetiological agents in human oesophageal cancers. The International Agency for Research on Cancer have designated toxins derived from F. moniliforme as group 2B (possibly carcinogenic to humans). Fumonisins are hepatotoxic, nephrotoxic, atherogenic, immunosuppressive and embryotoxic in experimental animal systems. Methods of detoxifying fumonisin-contaminated foods are required. Fumonisins have potent, apparently specific, inhibitory effects on sphingolipid biosynthesis and as such are valuable in studies of the complex biochemical events involved in sphingolipid metabolism and function. Fumonisins may serve as templates for therapeutic agents for treating diseases related to sphingolipid turnover (lysosomal storage disease), such as Farber's disease. PMID- 9876269 TI - Australian Aboriginal child health. AB - There have been substantial improvements in the health of Australian Aboriginal children over the past 2 decades. These include lower infant and toddler mortality rates and a significant decline in rates of hospitalization for conditions such as gastro-enteritis and lower respiratory tract infection. In addition, the degree of illness among these children on presentation is now generally much less severe than previously. There is evidence also of some improvement in birthweight, growth and nutritional status over the past 20 years. Incidence rates of infections among Aboriginal children, however, are still much higher than among their non-Aboriginal counterparts and much of this is due to unsatisfactory standards of living and community and personal hygiene. This is aggravated by widespread inadequate infrastructures for providing better housing, water supplies, solid and liquid waste disposal and the provision of regular, clean and nutritious food supplies in Aboriginal communities. These issues and more effective and culturally acceptable methods of disease prevention and health promotion are now being accorded high priority. But serious concerns remain about early Aboriginal "lifestyles" that may have important implications for health and mortality patterns among Aborigines during young to middle-age adult life. These include proneness to non-insulin-dependent diabetes mellitus, hypertension, cardiovascular disease, particularly ischaemic heart disease, and stroke which are likely to have their origins in childhood. The recent increase in rates of motor vehicle accidents, sometimes fatal, homicide and suicide, and the increasing rate of tobacco smoking and the use of addictive drugs, including the sniffing of petrol, glue and other volatile substances, is cause for serious concern for the future health and well-being of Aboriginal youth and their families. PMID- 9876271 TI - Japanese encephalitis. PMID- 9876270 TI - Ethnic health matters. PMID- 9876272 TI - Neonatology in developing countries: problems, practices and prospects. PMID- 9876273 TI - Iron and infection in the tropics: paediatric clinical correlates. AB - Iron deficiency is prevalent in children worldwide. Programmes of presumptive therapy, mass supplementation and food fortification have been introduced in many countries. The continuing unresolved debate over the interaction of iron and infection in the clinical setting indicates the need for firm guidelines for these practices. Iron overload is associated with increased susceptibility to certain infections, although the exact mechanisms may vary with the main pathology. Iron treatment has been associated with acute exacerbations of infection, in particular malaria. In Papua New Guinea parenteral iron was associated with increased rates of malaria and increased morbidity due to respiratory disease in infants but not in school children. Several subsequent studies in Africa using oral iron showed deleterious effects. In most instances cited, immunity was compromised, and therapeutic doses of oral iron were used. Knowledge of malarial endemicity, immunity with respect to age and the prevalence of haemoglobinopathies is important in planning interventions. A fine balance needs to be struck in the timing and dose of oral iron if informed recommendations are to be made. In parallel with supplementation studies, the effects of iron chelation on infection are being reported increasingly. Such therapy is clearly protective against malaria and some other infections but may predispose to fungal and Yersinia infections. PMID- 9876274 TI - Vitamin D deficiency rickets in developing countries. PMID- 9876275 TI - Flavianate, an amino acid precipitant, is a competitive inhibitor of trypsin at pH 3.0. AB - Textile dyes bind to proteins leading to selective co-precipitation of a complex involving one protein molecule and more than one dye molecule of opposite charge in acid solutions, in a process of reversible denaturation that can be utilized for protein fractionation. In order to understand what occurs before the co precipitation, a kinetic study using bovine beta-trypsin and sodium flavianate was carried out based on reaction progress curve techniques. The experiments were carried out using alpha-CBZ-L-Lys-p-nitrophenyl ester as substrate which was added to 50 mM sodium citrate buffer, pH 3.0, containing varying concentrations of beta-trypsin and dye. The reaction was recorded spectrophotometrically at 340 nm for 30 min, and the families of curves obtained were analyzed simultaneously by fitting integrated Michaelis-Menten equations. The dye used behaved as a competitive inhibitor of trypsin at pH 3.0, with Ki = 99 microM; kinetic parameters for the substrate hydrolysis were: Km = 32 microM, and kcat = 0.38/min. The competitive character of the inhibition suggests a specific binding of the first dye molecule to His-57, the only positively charged residue at the active site of the enzyme. PMID- 9876276 TI - Gender differences in the activities of aspirin-esterases in rat tissues. AB - The activities of aspirin (acetylsalicylic acid)-esterases were measured in several tissues (liver, kidney, adrenal glands, brain and serum) from adult male and female Wistar rats. In males, both aspirin-esterase I (assayed at pH 5.5) and II (assayed at pH 7.4) activities were higher in liver homogenates when compared to females (aspirin-esterase I: males 48.9 +/- 4.8 (N = 8) and females 29.3 +/- 4.2 (N = 8) nmol of salicylic acid formed min-1 mg protein-1; aspirin-esterase II: males 41.4 +/- 4.1 (N = 8) and females 26.1 +/- 4.5 (N = 8) nmol of salicylic acid formed min-1 mg protein-1, P < 0.001). In serum, enzyme activity was higher in females than in males (aspirin-esterase I: males 0.85 +/- 0.06 (N = 6) and females 1.18 +/- 0.11 (N = 6) nmol of salicylic acid formed min-1 mg protein-1, aspirin-esterase II: males 1.03 +/- 0.13 (N = 6) and females 1.34 +/- 0.11 (N = 6) nmol of salicylic acid formed min-1 mg protein-1, P < 0.001). In the other tissues assayed, no statistically significant difference between males and females was found. There were no statistically significant differences when the enzymes were assayed in different phases of the estrous cycle in liver and serum. These results show that the differences in aspirin-esterase activity observed between males and females are not due to the estrous cycle. The gender difference obtained in our study may indicate an involvement of gonadal hormones in the control of the hydrolysis of aspirin. This possibility is currently under investigation. PMID- 9876277 TI - Inhibition of Mayaro virus replication by prostaglandin A1 and B2 in Vero cells. AB - The effect of prostaglandins (PGA1 and PGB2) on the replication of Mayaro virus was studied in Vero cells. PGA1 and PGB2 antiviral activity was found to be dose dependent. However, while 10 micrograms/ml PGB2 inhibited virus yield by 60%, at the same dose PGA1 suppressed virus replication by more than 90%. SDS-PAGE analysis of [35S]-methionine-labelled proteins showed that PGA1 did not alter cellular protein synthesis. In infected cells, PGA1 slightly inhibited the synthesis of protein C, while drastically inhibiting the synthesis of glycoproteins E1 and E2. PMID- 9876278 TI - A single-step purification of bothropstoxin-1. AB - Bothrops venoms are complex mixtures of components with a wide range of biological activities. Among these substances, myotoxins have been investigated by several groups. Bothropstoxin-1 (Bthtx-1) is a phospholipase A2-like basic myotoxin from Bothrops jararacussu. The purification of this component involves two chromatographic steps. Although providing a pure material, the association of these two steps is time consuming and a single-step method using high performance chromatography media would be useful. In the present study, we describe a single step purification method for Bthtx-1. Bothrops jararacussu venom was dissolved in 1 ml buffer. After centrifugation, the supernatant was injected into a Resource-S cation exchange column connected to an FPLC system and eluted with a linear salt gradient. The complete procedure took 20 min, representing a considerable time gain when compared to a previously described method (Homsi-Brandenburgo MI et al. (1988) Toxicon, 26: 615-627). Bthtx-1 purity and identity, assessed by SDS-PAGE and N-terminal sequencing, resulted in a single band with a molecular mass of about 14 kDa and the expected sequence of the first 5 residues, S-L-F-E-L. Although the amount of protein purified after each run is lower than in the previously described method, we believe that this method may be useful for small scale purifications. PMID- 9876279 TI - Use of a cysteine proteinase from Carica candamarcensis as a protective agent during DNA extraction. AB - We describe the use of a plant cysteine proteinase isolated from latex of Carica candamarcensis as a protective agent during isolation of bacterial DNA following growth in culture of these cells. Between 100 to 720 units of proteinase (1 microgram = 6 units) afforded good DNA protection when incubated with various kinds of microorganisms. Agarose gel electrophoresis showed that the resulting DNA was similar in size to DNA preparations obtained by treatment with proteinase K. The viability of the resulting material was checked by PCR amplification using species-specific primers. After standing at room temperature (25 degrees C) for 35 days, the enzyme lost 10% of its initial activity. The enzyme stability and good yield of DNA suggest the use of this proteinase as an alternative to proteinase K. PMID- 9876280 TI - Gastric emptying in rats with acetaminophen-induced hepatitis. AB - The objective of this work was to study the gastric emptying (GE) of liquids in fasted and sucrose-fed rats with toxic hepatitis induced by acetaminophen. The GE of three test meals (saline, glucose and mayonnaise) was evaluated in Wistar rats. For each meal, the animals were divided into two groups (N = 24 each). Group I was fed a sucrose diet throughout the experiment (66 h) while group II was fasted. Forty-two hours after the start of the experiment, each group was divided into two subgroups (N = 12 each). Subgroup A received a placebo and subgroup B was given acetaminophen (1 g/kg). Twenty-four hours later, the GE of the three test meals was assessed and blood samples were collected to measure the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and acetaminophen. In group IB, the mean AST and ALT values were 515 and 263 IU/l, respectively, while for group IIB they were 4014 and 2472 IU/l, respectively. The mean serum acetaminophen levels were higher in group IIB (120 micrograms/ml) than in group IB (87 micrograms/ml). The gastric retention values were significantly higher in group IIB than in group IIA for all three test meals: saline, 51 vs 35%; glucose, 52 vs 38% and mayonnaise, 51 vs 29% (median values). The correlation between gastric retention and AST levels was significant (P < 0.05) for group IIB for the three test meals: r = 0.73, 0.67 and 0.68 for saline, glucose and mayonnaise, respectively. We conclude that GE is altered in rats with hepatic lesions induced by acetaminophen, and that these alterations may be related to the liver cell necrosis caused by the drug. PMID- 9876281 TI - Detection of adrenocortical autoantibodies in Addison's disease with a peroxidase labelled protein A technique. AB - Adrenocortical autoantibodies (ACA), present in 60-80% of patients with idiopathic Addison's disease, are conventionally detected by indirect immunofluorescence (IIF) on frozen sections of adrenal glands. The large-scale use of IIF is limited in part by the need for a fluorescence microscope and the fact that histological sections cannot be stored for long periods of time. To circumvent these restrictions we developed a novel peroxidase-labelled protein A (PLPA) technique for the detection of ACA in patients with Addison's disease and compared the results with those obtained with the classical IIF assay. We studied serum samples from 90 healthy control subjects and 22 patients with Addison's disease, who had been clinically classified into two groups: idiopathic (N = 13) and granulomatous (N = 9). ACA-PLPA were detected in 10/22 (45%) patients: 9/13 (69%) with the idiopathic form and 1/9 (11%) with the granulomatous form, whereas ACA-IIF were detected in 11/22 patients (50%): 10/13 (77%) with the idiopathic form and 1/9 (11%) with the granulomatous form. Twelve of the 13 idiopathic addisonians (92%) were positive for either ACA-PLPA or ACA-IIF, but only 7 were positive by both methods. In contrast, none of 90 healthy subjects was found to be positive for ACA. Thus, our study shows that the PLPA-based technique is useful, has technical advantages over the IIF method (by not requiring the use of a fluorescence microscope and by permitting section storage for long periods of time). However, since it is only 60% concordant with the ACA-IIF method, it should be considered complementary instead of an alternative method to IIF for the detection of ACA in human sera. PMID- 9876283 TI - Methylmercury intoxication and histochemical demonstration of NADPH-diaphorase activity in the striate cortex of adult cats. AB - The effects of methylmercury (MeHg) on histochemical demonstration of the NADPH diaphorase (NADPH-d) activity in the striate cortex were studied in 4 adult cats. Two animals were used as control. The contaminated animals received 50 ml milk containing 0.42 microgram MeHg and 100 g fish containing 0.03 microgram MeHg daily for 2 months. The level of MeHg in area 17 of intoxicated animals was 3.2 micrograms/g wet weight brain tissue. Two cats were perfused 24 h after the last dose (group 1) and the other animals were perfused 6 months later (group 2). After microtomy, sections were processed for NADPHd histochemistry procedures using the malic enzyme method. Dendritic branch counts were performed from camera lucida drawings for control and intoxicated animals (N = 80). Average, standard deviation and Student t-test were calculated for each data group. The concentrations of mercury (Hg) in milk, fish and brain tissue were measured by acid digestion of samples, followed by reduction of total Hg in the digested sample to metallic Hg using stannous chloride followed by atomic fluorescence analysis. Only group 2 revealed a reduction of the neuropil enzyme activity and morphometric analysis showed a reduction in dendritic field area and in the number of distal dendrite branches of the NADPHd neurons in the white matter (P < 0.05). These results suggest that NADPHd neurons in the white matter are more vulnerable to the long-term effects of MeHg than NADPHd neurons in the gray matter. PMID- 9876282 TI - Characterization of Plasmodium falciparum glutamate dehydrogenase-soluble antigen. AB - The major aim of this study was to characterize a soluble Plasmodium falciparum antigen from the plasma of malaria-infected humans and Plasmodium falciparum culture supernatants, using immunoabsorbent techniques and Western blotting. An Mr 60-kDa protein was isolated from the plasma of patients with Plasmodium falciparum malaria by affinity chromatography using rabbit anti-Proteus spp GDH(NADP+) serum as ligand. This protein, present in plasma of patients with acute Plasmodium falciparum infection, in Plasmodium falciparum culture supernatants, and in immune complexes, was tested with Plasmodium falciparum malaria hyperimmune serum from patients living in hyperendemic areas and rabbit anti-Proteus spp GDH(NADP+) serum prepared in the laboratory. In this report, we describe the results of a study showing that parasite GDH(NADP+) can be used to detect the presence of Plasmodium falciparum. It appears that this technique permits the chromatographic detection of a Plasmodium falciparum excretion antigen that may be used in the production of monoclonal antibodies to improve immunodiagnostic assays for the detection of antigenemia, and opens the possibility of its use as a non-microscopic screening method. PMID- 9876284 TI - Antioxidant and free radical scavenging effects in extracts of the medicinal herb Achyrocline satureioides (Lam.) DC. ("marcela"). AB - Achyrocline satureioides (Lam.) DC. (Compositae) is a medicinal herb used in Argentina, Uruguay, Brazil and Paraguay for its choleretic, antispasmodic and hepatoprotective properties. The presence of the flavonoid quercetin and its derivatives, and of different phenolic acids such as caffeic, chlorogenic and isochlorogenic acids in the aerial parts of this plant has led us to study the antioxidant activity of its extracts using different bioassays. The inhibition of luminol-enhanced chemiluminescence by the aqueous and methanolic extracts was used to show that their total reactive antioxidant potential index (TRAP; in microM Trolox equivalents) was 91.0 +/- 15.4 and 128.1 +/- 20.1 microM, respectively, while the total antioxidant reactivity index (TAR) was calculated to be 1537 +/- 148 and 1910 +/- 171 microM. Only the methanolic extract was capable of reducing iron (II)-dependent DNA damage. Lipid peroxidation was assessed by two different methods. The aqueous extract reduced hydroperoxide initiated chemiluminescence in rat liver homogenates at all concentrations in a dose-dependent manner, with a calculated IC50 = 225 micrograms/ml, while the methanolic extract was only effective at higher concentrations (100 and 1000 micrograms/ml). Both aqueous and methanolic extracts were capable of reducing the production of thiobarbituric acid reactive substances (TBARS) in rat liver homogenates, with an IC50 > 1000 micrograms/ml. The results obtained suggest that the extracts of A. satureioides possess significant free radical scavenging and antioxidant activity in vitro, a fact that should encourage future in vivo studies. PMID- 9876285 TI - Transgenic animal models for the functional analysis of vasoactive peptides. AB - The interplay of vasoactive peptide systems is an essential determinant of blood pressure regulation in mammals. While the endothelin and the renin-angiotensin systems raise blood pressure by inducing vasoconstriction and sodium retention, the kallikrein-kinin and the natriuretic-peptide systems reduce arterial pressure by eliciting vasodilatation and natriuresis. Transgenic technology has proven to be very useful for the functional analysis of vasoactive peptide systems. As an outstanding example, transgenic rats overexpressing the mouse Ren-2 renin gene in several tissues become extremely hypertensive. Several other transgenic rat and mouse strains with genetic modifications of components of the renin-angiotensin system have been developed in the past decade. Moreover, in recent years gene targeting technology was employed to produce mouse strains lacking these proteins. The established animal models as well as the main insights gained by their analysis are summarized in this review. PMID- 9876286 TI - Endogenous vasopressin and the central control of heart rate during dynamic exercise. AB - The present article contains a brief review on the role of vasopressinergic projections to the nucleus tractus solitarii in the genesis of reflex bradycardia and in the modulation of heart rate control during exercise. The effects of vasopressin on exercise tachycardia are discussed on the basis of both the endogenous peptide content changes and the heart rate response changes observed during running in sedentary and trained rats. Dynamic exercise caused a specific vasopressin content increase in dorsal and ventral brainstem areas. In accordance, rats pretreated with the peptide or the V1 blocker into the nucleus tractus solitarii showed a significant potentiation or a marked blunting of the exercise tachycardia, respectively, without any change in the pressure response to exercise. It is proposed that the long-descending vasopressinergic pathway to the nucleus tractus solitarii serves as one link between the two main neural controllers of circulation, i.e., the central command and feedback control mechanisms driven by the peripheral receptors. Therefore, vasopressinergic input could contribute to the adjustment of heart rate response (and cardiac output) to the circulatory demand during exercise. PMID- 9876287 TI - Molecular biology of the kallikrein-kinin system: from structure to function. AB - The participation of the kallikrein-kinin system, comprising the serine proteases kallikreins, the protein substrates kininogens and the effective peptides kinins, in some pathological processes like hypertension and cardiovascular diseases is still a matter of controversy. The use of different experimental set-ups in concert with the development of potent and specific inhibitors and antagonists for the system has highlighted its importance but the results still lack conclusivity. Over the last few years, transgenic and gene-targeting technologies associated with molecular biology tools have provided specific information about the elusive role of the kallikrein-kinin system in the control of blood pressure and electrolyte homeostasis. cDNA and genomic sequences for kinin receptors B2 and B1 from different species were isolated and shown to encode G-protein-coupled receptors and the structure and pharmacology of the receptors were characterized. Transgenic animals expressing an overactive kallikrein-kinin system were established to study the cardiovascular effects of these alterations and the results of these investigations further corroborate the importance of this system in the maintenance of normal blood pressure. Knockout animals for B2 and B1 receptors are available and their analysis also points to the role of these receptors in cardiovascular regulation and inflammatory processes. In this paper the most recent and relevant genetic animal models developed for the study of the kallikrein-kinin system are reviewed, and the advances they brought to the understanding of the biological role of this system are discussed. PMID- 9876288 TI - Antihypertensive effects of angiotensin-(1-7). AB - Accumulating evidence suggests that angiotensin-(1-7)(Ang-(1-7)) is an important component of the renin-angiotensin system and that the actions of the peptide may either contribute to or oppose those of Ang II. Ang-(1-7) can be converted directly from Ang I bypassing prerequisite formation of Ang II. Formation of Ang (1-7) is under the control of at least three endopeptidases depending on the tissue compartment and include neprilysin, thimet oligopeptidase and prolyl oligopeptidase. Both neprilysin and thimet oligopeptidase are also involved in the metabolism of bradykinin and the atrial natriuretic peptide. Moreover, recent studies suggest that in addition to Ang I and bradykinin, Ang-(1-7) is an endogenous substrate for angiotensin converting enzyme. These enzymatic pathways may contribute to a complex relationship between the hypertensive actions of Ang II and various vasodepressor peptides from either the renin-angiotensin system or other peptide systems. Ang-(1-7) is devoid of the vasoconstrictor, central pressor, or thirst-stimulating actions associated with Ang II. In fact, new findings reveal depressor, vasodilator, and antihypertensive actions that may be more apparent in hypertensive animals or humans. Thus, Ang-(1-7) may oppose the actions of Ang II directly or as a result of increasing prostaglandins or nitric oxide. In this review, we examine the mechanisms by which Ang-(1-7) may contribute to cardiovascular regulation. PMID- 9876289 TI - Baroreflex control of sympathetic activity in experimental hypertension. AB - The arterial baroreceptor reflex system is one of the most powerful and rapidly acting mechanisms for controlling arterial pressure. The purpose of the present review is to discuss data relating sympathetic activity to the baroreflex control of arterial pressure in two different experimental models: neurogenic hypertension by sinoaortic denervation (SAD) and high-renin hypertension by total aortic ligation between the renal arteries in the rat. SAD depresses baroreflex regulation of renal sympathetic activity in both the acute and chronic phases. However, increased sympathetic activity (100%) was found only in the acute phase of sinoaortic denervation. In the chronic phase of SAD average discharge normalized but the pattern of discharges was different from that found in controls. High-renin hypertensive rats showed overactivity of the renin angiotensin system and a great depression of the baroreflexes, comparable to the depression observed in chronic sinoaortic denervated rats. However, there were no differences in the average tonic sympathetic activity or changes in the pattern of discharges in high-renin rats. We suggest that the difference in the pattern of discharges may contribute to the increase in arterial pressure lability observed in chronic sinoaortic denervated rats. PMID- 9876290 TI - Effect of selective angiotensin antagonists on the antidiuresis produced by angiotensin-(1-7) in water-loaded rats. AB - In the present study we evaluated the nature of angiotensin receptors involved in the antidiuretic effect of angiotensin-(1-7) (Ang-(1-7)) in water-loaded rats. Water diuresis was induced in male Wistar rats weighing 280 to 320 g by water load (5 ml/100 g body weight by gavage). Immediately after water load the rats were treated subcutaneously with (doses are per 100 g body weight): 1) vehicle (0.05 ml 0.9% NaCl); 2) graded doses of 20, 40 or 80 pmol Ang-(1-7); 3) 200 nmol Losartan; 4) 200 nmol Losartan combined with 40 pmol Ang-(1-7); 5) 1.1 or 4.4 nmol A-779; 6) 1.1 nmol A-779 combined with graded doses of 20, 40 or 80 pmol Ang (1-7); 7) 4.4 nmol A-779 combined with graded doses of 20, 40 or 80 pmol Ang-(1 7); 8) 95 nmol CGP 42112A, or 9) 95 nmol CGP 42112A combined with 40 pmol Ang-(1 7). The antidiuretic effect of Ang-(1-7) was associated with an increase in urinary Na+ concentration, an increase in urinary osmolality and a reduction in creatinine clearance (CCr: 0.65 +/- 0.04 ml/min vs 1.45 +/- 0.18 ml/min in vehicle-treated rats, P < 0.05). A-779 and Losartan completely blocked the effect of Ang-(1-7) on water diuresis (2.93 +/- 0.34 ml/60 min and 3.39 +/- 0.58 ml/60 min, respectively). CGP 42112A, at the dose used, did not modify the antidiuretic effect of Ang-(1-7). The blockade produced by Losartan was associated with an increase in CCr and with an increase in sodium and water excretion as compared with Ang-(1-7)-treated rats. When Ang-(1-7) was combined with A-779 there was an increase in CCr and natriuresis and a reduction in urine osmolality compared with rats treated with Ang-(1-7) alone. The observation that both A-779, which does not bind to AT1 receptors, and Losartan blocked the effect of Ang-(1-7) suggests that the kidney effects of Ang-(1-7) are mediated by a non-AT1 angiotensin receptor that is recognized by Losartan. PMID- 9876291 TI - Role of non-nitric oxide non-prostaglandin endothelium-derived relaxing factor(s) in bradykinin vasodilation. AB - The most conspicuous effect of bradykinin following its administration into the systemic circulation is a transient hypotension due to vasodilation. In the present study most of the available evidence regarding the mechanisms involved in bradykinin-induced arterial vasodilation is reviewed. It has become firmly established that in most species vasodilation in response to bradykinin is mediated by the release of endothelial relaxing factors following the activation of B2-receptors. Although in some cases the action of bradykinin is entirely mediated by the endothelial release of nitric oxide (NO) and/or prostacyclin (PGI2), a large amount of evidence has been accumulated during the last 10 years indicating that a non-NO/PGI2 factor accounts for bradykinin-induced vasodilation in a wide variety of perfused vascular beds and isolated small arteries from several species including humans. Since the effect of the non-NO/PGI2 endothelium derived relaxing factor is practically abolished by disrupting the K+ electrochemical gradient together with the fact that bradykinin causes endothelium-dependent hyperpolarization of vascular smooth muscle cells, the action of such factor has been attributed to the opening of K+ channels in these cells. The pharmacological characteristics of these channels are not uniform among the different blood vessels in which they have been examined. Although there is some evidence indicating a role for KCa or KV channels, our findings in the mesenteric bed together with other reports indicate that the K+ channels involved do not correspond exactly to any of those already described. In addition, the chemical identity of such hyperpolarizing factor is still a matter of controversy. The postulated main contenders are epoxyeicosatrienoic acids or endocannabinoid agonists for the CB1-receptors. Based on the available reports and on data from our laboratory in the rat mesenteric bed, we conclude that the NO/PGI2-independent endothelium-dependent vasodilation induced by BK is unlikely to involve a cytochrome P450 arachidonic acid metabolite or an endocannabinoid agonist. PMID- 9876292 TI - Accuracy of replication in the polymerase chain reaction. Comparison between Thermotoga maritima DNA polymerase and Thermus aquaticus DNA polymerase. AB - For certain applications of the polymerase chain reaction (PCR), it may be necessary to consider the accuracy of replication. The breakthrough that made PCR user friendly was the commercialization of Thermus aquaticus (Taq) DNA polymerase, an enzyme that would survive the high temperatures needed for DNA denaturation. The development of enzymes with an inherent 3' to 5' exonuclease proofreading activity, lacking in Taq polymerase, would be an improvement when higher fidelity is needed. We used the forward mutation assay to compare the fidelity of Taq polymerase and Thermotoga maritima (ULTMA) DNA polymerase, an enzyme that does have proofreading activity. We did not find significant differences in the fidelity of either enzyme, even when using optimal buffer conditions, thermal cycling parameters, and number of cycles (0.2% and 0.13% error rates for ULTMA and Taq, respectively, after reading about 3,000 bases each). We conclude that for sequencing purposes there is no difference in using a DNA polymerase that contains an inherent 3' to 5' exonuclease activity for DNA amplification. Perhaps the specificity and fidelity of PCR are complex issues influenced by the nature of the target sequence, as well as by each PCR component. PMID- 9876293 TI - Two variants of HIV-1 B serotype are transmitted heterosexually in Sao Paulo, Brazil. AB - HIV-1 variability may have an important impact on transmission and pathogenicity. Better characterization of the HIV epidemic in Brazil is necessary for the development of vaccine trials in this country. We analyzed sera from 108 HIV-1 infected volunteers from Sao Paulo City to determine serotype and reactivity for V3 motifs of HIV in this population, and the relationship to transmission mode. We concluded that the HIV-1 B serotype is frequent among heterosexually infected women, even in the absence of anal sex, and that two major V3 motifs, GPGR and GWGR, had similar prevalence among women (48% and 52%, respectively) and men (56% and 44%, respectively). We also observed an equal distribution of these strains regardless of their CD4+ T cell counts, clinical status, and mode of transmission. Even though V3 serology for HIV-1 subtyping is an inexpensive tool for use in developing countries, additional methods, such as heteroduplex mobility assay and direct DNA sequencing, should be included to determine HIV-1 genetic diversity. PMID- 9876294 TI - Post-exercise changes in blood pressure, heart rate and rate pressure product at different exercise intensities in normotensive humans. AB - To evaluate the effect of exercise intensity on post-exercise cardiovascular responses, 12 young normotensive subjects performed in a randomized order three cycle ergometer exercise bouts of 45 min at 30, 50 and 80% of VO2peak, and 12 subjects rested for 45 min in a non-exercise control trial. Blood pressure (BP) and heart rate (HR) were measured for 20 min prior to exercise (baseline) and at intervals of 5 to 30 (R5-30), 35 to 60 (R35-60) and 65 to 90 (R65-90) min after exercise. Systolic, mean, and diastolic BP after exercise were significantly lower than baseline, and there was no difference between the three exercise intensities. After exercise at 30% of VO2peak, HR was significantly decreased at R35-60 and R65-90. In contrast, after exercise at 50 and 80% of VO2peak, HR was significantly increased at R5-30 and R35-60, respectively. Exercise at 30% of VO2peak significantly decreased rate pressure (RP) product (RP = HR x systolic BP) during the entire recovery period (baseline = 7930 +/- 314 vs R5-30 = 7150 +/ 326, R35-60 = 6794 +/- 349, and R65-90 = 6628 +/- 311, P < 0.05), while exercise at 50% of VO2peak caused no change, and exercise at 80% of VO2peak produced a significant increase at R5-30 (7468 +/- 267 vs 9818 +/- 366, P < 0.05) and no change at R35-60 or R65-90. Cardiovascular responses were not altered during the control trial. In conclusion, varying exercise intensity from 30 to 80% of VO2peak in young normotensive humans did not influence the magnitude of post exercise hypotension. However, in contrast to exercise at 50 and 80% of VO2peak, exercise at 30% of VO2peak decreased post-exercise HR and RP. PMID- 9876295 TI - Renal dysfunction in patients with sickle cell anemia or sickle cell trait. AB - Patients with sickle cell anemia (Hb SS) or sickle cell trait (Hb AS) may present several types of renal dysfunction; however, comparison of the prevalence of these abnormalities between these two groups and correlation with the duration of disease in a large number of patients have not been thoroughly investigated. In a cross-sectional study using immunoenzymometric assays to measure tubular proteinuria, microalbuminuria, measurement of creatinine clearance, urinary osmolality and analysis of urine sediment, we evaluated glomerular and tubular renal function in 106 adults and children with Hb SS (N = 66) or Hb AS (N = 40) with no renal failure (glomerular filtration rate (GFR) > 85 ml/min). The percentage of individuals with microalbuminuria was higher among Hb SS than among Hb AS patients (30 vs 8%, P < 0.0001). The prevalence of microhematuria was similar in both groups (26 vs 30%, respectively). Increased urinary levels of retinol-binding protein or beta 2-microglobulin were detected in only 3 Hb SS and 2 Hb AS patients. Urinary osmolality was reduced in patients with Hb SS or with Hb AS; however, it was particularly evident in Hb SS patients older than 15 years (median = 393 mOsm/kg, range = 366-469) compared with Hb AS patients (median = 541 mOsm/kg, range = 406-722). Thus, in addition to the frequently reported early reduction of urinary osmolality and increased GFR, nondysmorphic hematuria was found in 26 and 30% of patients with Hb SS or Hb AS, respectively. Microalbuminuria is an important marker of glomerular injury in patients with Hb SS and may also be demonstrated in some Hb AS individuals. Significant proximal tubular dysfunction is not a common feature in Hb SS and Hb AS population at this stage of the disease (i.e., GFR > 85 ml/min). PMID- 9876296 TI - Serodiagnosis of Helicobacter pylori infection by Cobas Core ELISA in adults from Minas Gerais, Brazil. AB - We evaluated the accuracy of a 2nd generation ELISA to detect Helicobacter pylori infection in adults from a developing country in view of variations in sensitivity and specificity reported for different populations. We studied 97 non consecutive patients who underwent endoscopy for evaluation of dispeptic symptoms. The presence of H. pylori was determined in antral biopsy specimens by culture, by the preformed urease test and in carbolfuchsin-stained smears. Patients were considered to be H. pylori positive if at least two of the three tests presented a positive result or if the culture was positive, and negative if the three tests were negative. Sixty-five adults (31 with peptic ulcer) were H. pylori positive and 32 adults were H. pylori negative. Antibodies were detected by Cobas Core anti-H. pylori EIA in 62 of 65 H. pylori-positive adults and in none of the negative adults. The sensitivity, specificity and positive and negative predictive values of the test were 95.4, 100, 100 and 91.4%, respectively. The Cobas Core anti-H. pylori EIA presented high sensitivity and specificity when employed for a population in Brazil, permitting the use of the test both to confirm the clinical diagnosis and to perform epidemiologic surveys. PMID- 9876297 TI - Effect of metabolic acidosis on renal tubular sodium handling in rats as determined by lithium clearance. AB - Systemic metabolic acidosis is known to cause a decrease in salt and water reabsorption by the kidney. We have used renal lithium clearance to investigate the effect of chronic, NH4Cl-induced metabolic acidosis on the renal handling of Na+ in male Wistar-Hannover rats (200-250 g). Chronic acidosis (pH 7.16 +/- 0.13) caused a sustained increase in renal fractional Na+ excretion (267.9 +/- 36.4%), accompanied by an increase in fractional proximal (113.3 +/- 3.6%) and post proximal (179.7 +/- 20.2%) Na+ and urinary K+ (163.4 +/- 5.6%) excretion when compared to control and pair-fed rats. These differences occurred in spite of an unchanged creatinine clearance and Na+ filtered load. A lower final body weight was observed in the acidotic (232 +/- 4.6 g) and pair-fed (225 +/- 3.6 g) rats compared to the controls (258 +/- 3.7 g). In contrast, there was a significant increase in the kidney weights of acidotic rats (1.73 +/- 0.05 g) compared to the other experimental groups (control, 1.46 +/- 0.05 g; pair-fed, 1.4 +/- 0.05 g). We suggest that altered renal Na+ and K+ handling in acidotic rats may result from a reciprocal relationship between the level of metabolism in renal tubules and ion transport. PMID- 9876298 TI - Short duration of neutralizing antibody titers after pre-exposure rabies vaccination with suckling mouse brain vaccine. AB - The human anti-rabies pre-exposure treatment currently used in Brazil, employing a 1-ml dose of suckling mouse brain vaccine (SMBV) administered on days 0, 2, 4 and 28, was compared to an alternative treatment with two 1 ml-doses on day 0, and one 1 ml-dose injected on days 7 and 21. The latter induced higher virus neutralizing antibody (VNA) titers on day 21. Both Brazilian rabies vaccines produced with PV or CVS rabies virus strains were tested. Two additional volunteer vaccine groups, receiving the pre-exposure and the abbreviated post exposure schedules recommended by the WHO using cell-culture vaccine (CCV) produced with PM rabies virus strain, were included as reference. The VNA were measured against both PV and CVS strains on days 21, 42 and 180 by the cell culture neutralization microtest. The PV-SMBV elicited higher seroconversion rates and VNA by day 21 than the CVS-SMBV. Both, however, failed to induce a long term immunity, since VNA titers were < 0.5 IU/ml on day 180, regardless of the schedule used. Cell-culture vaccine always elicited very high VNA on all days of collection. When serum samples from people receiving mouse brain tissue were titrated against the PV and CVS strains, the VNA obtained were similar, regardless of the vaccinal strain and the virus used in the neutralization test. These results contrast with those obtained with sera from people receiving PM CCV, whose VNA were significantly higher when tested against the CVS strain. PMID- 9876299 TI - Thyroid hormone regulates protein expression in C6 glioma cells. AB - Thyroid hormone (T3) is essential to normal brain development. Previously, we have shown that T3 induces cerebellar astrocyte proliferation. This effect is accompanied by alteration in glial fibrillary acidic protein (GFAP) and fibronectin organization. In the present study, we report that the C6 glioma cell line, which expresses GFAP and is classified as an undifferentiated astrocytic cell type, is a target for T3 action. The C6 monolayers were treated with 50 nM T3 for 3 days, after which the cells were maintained for 2 days without medium changes. In C6 cells, T3 induced the expression of proteins of 107, 73 and 62 kDa. The hormone also up-regulated protein bands of 100 (+50%), 37 (+50%) and 25.5 kDa (+50%) and down-regulated proteins of 94 (-100%), 86.5 (-100%), 68 ( 100%), 60 (-100%), 54 (-33%), 51 (-33%) and 43.5 kDa (-33%). We suggest, on the basis of molecular mass, that the 54-, 51- and 43.5-kDa proteins could be the cytoskeletal proteins vimentin, GFAP and actin, respectively. The down-regulation of these proteins may be involved in the effects of thyroid hormone on C6 differentiation. PMID- 9876300 TI - Identification of the main generator source of longitudinal muscle contraction in the earthworm ventral nerve cord. AB - The main generator source of a longitudinal muscle contraction was identified as an M (mechanical-stimulus-sensitive) circuit composed of a presynaptic M-1 neuron and a postsynaptic M-2 neuron in the ventral nerve cord of the earthworm, Amynthas hawayanus, by simultaneous intracellular response recording and Lucifer Yellow-CH injection with two microelectrodes. Five-peaked responses were evoked in both neurons by a mechanical, but not by an electrical, stimulus to the mechanoreceptor in the shaft of a seta at the opposite side of an epidermis muscle-nerve-cord preparation. This response was correlated to 84% of the amplitude, 73% of the rising rate and 81% of the duration of a longitudinal muscle contraction recorded by a mechanoelectrical transducer after eliminating the other possible generator sources by partitioning the epidermis-muscle piece of this preparation. The pre- and postsynaptic relationship between these two neurons was determined by alternately stimulating and recording with two microelectrodes. Images of the Lucifer Yellow-CH-filled M-1 and M-2 neurons showed that both of them are composed of bundles of longitudinal processes situated on the side of the nerve cord opposite to stimulation. The M-1 neuron has an afferent process (A1) in the first nerve at the stimulated side of this preparation and the M-2 neuron has two efferent processes (E1 and E3) in the first and third nerves at the recording side where their effector muscle cell was identified by a third microelectrode. PMID- 9876301 TI - Facilitation of the main generator source of earthworm muscle contraction by a peripheral neuron. AB - A constant facilitation of responses evoked in the earthworm muscle contraction generator neurons by responses evoked in the neurons of its peripheral nervous system was demonstrated. It is based on the proposal that these two responses are bifurcations of an afferent response evoked by the same peripheral mechanical stimulus but converging again on this central neuron. A single-peaked generator response without facilitation was demonstrated by sectioning the afferent route of the peripheral facilitatory modulatory response, or conditioning response (CR). The multipeaked response could be restored by restimulating the sectioned modulatory neuron with an intracellular substitutive conditioning stimulus (SCS). These multipeaked responses were proposed to be the result of reverberating the original single peaked unconditioned response (UR) through a parallel (P) neuronal circuit which receives the facilitation of the peripheral modulatory neuron. This peripheral modulatory neuron was named "Peri-Kastchen" (PK) neuron because it has about 20 peripheral processes distributed on the surface of a Kastchen of longitudinal muscle cells on the body wall of this preparation as revealed by the Lucifer Yellow-CH-filling method. PMID- 9876302 TI - Persistent attenuation and enhancement of the earthworm main muscle contraction generator response induced by repeated stimulation of a peripheral neuron. AB - Responses evoked in the earthworm, Amynthas hawayanus, main muscle contraction generator M-2 (postsynaptic mechanical-stimulus-sensitive) neuron by threshold mechanical stimuli in 2-s intertrial intervals (ITI) were used as the control or unconditioned responses (UR). Their attenuation induced by decreasing these intervals in non-associative conditioning and their enhancement induced by associating the unconditioned stimuli (US) to a train of short (0.1 s) hyperpolarizing electrical substitutive conditioning stimuli (SCS) in the Peri Kastchen (PK) neuron were measured in four parameters, i.e., peak numbers (N) and amplitude (AMP) averaged from 120 responses, sum of these amplitudes (sigma AMP) and the highest peak amplitude (V) over a period of 4 min. Persistent attenuation similar to habituation was induced by decreasing the control ITI to 0.5 s and 2.0 s in non-associative conditioning within less than 4 min. Dishabituation was induced by randomly pairing one of these habituated US to an electrical stimulus in the PK neuron. All four parameters of the UR were enhanced by forward (SCS US), but not backward (US-SCS), association of the US with 25, 100 and 250-Hz trains of SCS with 40-ms interstimulus intervals (ISI) for 4 min and persisted for another 4 min after turning off the SCS. The enhancement of these parameters was proportional to the SCS frequencies in the train. No UR was evoked by the SCS when the US was turned off after 4 min of classical conditioning. PMID- 9876303 TI - Gap effect and reaction time distribution: simple vs choice manual responses. AB - It is well known that saccadic reaction times (SRT) are reduced when the target is preceded by the offset of the fixation point (FP)--the gap effect. Some authors have proposed that the FP offset also allows the saccadic system to generate a separate population of SRT, the express saccades. Nevertheless, there is no agreement as to whether the gap effect and express responses are also present for manual reaction times (MRT). We tested the gap effect and the MRT distribution in two different conditions, i.e., simple and choice MRT. In the choice MRT condition, subjects need to identify the side of the stimulus and to select the appropriate response, while in the simple MRT these stages are not necessary. We report that the gap effect was present in both conditions (22 ms for choice MRT condition; 15 ms for simple MRT condition), but, when analyzing the MRT distributions, we did not find any clear evidence for express manual responses. The main difference in MRT distribution between simple and choice conditions was a shift towards shorter values for simple MRT. PMID- 9876305 TI - Endothelium-dependent vasodilation in response to Pseudomonas aeruginosa lipopolysaccharide: an in vitro study on canine arteries. AB - Early systemic arterial hypotension is a common clinical feature of Pseudomonas septicemia. To determine if Pseudomonas aeruginosa endotoxin induces the release of endothelium-derived nitric oxide (EDNO), an endogenous nitrovasodilator, segments of canine femoral, renal, hepatic, superior mesenteric, and left circumflex coronary arteries were suspended in organ chambers (physiological salt solution, 95% O2/5% CO2, pH 7.4, 37 degrees C) to measure isometric force. In arterial segments contracted with 2 microM prostaglandin F2 alpha, Pseudomonas endotoxin (lipopolysaccharide (LPS) serotype 10(Habs) from Pseudomonas aeruginosa (0.05 to 0.50 mg/ml) induced concentration-dependent relaxation of segments with endothelium (P < 0.05) but no significant change in tension of arteries without endothelium. Endothelium-dependent relaxation in response to Pseudomonas LPS occurred in the presence of 1 microM indomethacin, but could be blocked in the coronary artery with 10 microM NG-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of nitric oxide synthesis from L-arginine. The inhibitory effect of L NMMA on LPS-mediated vasorelaxation of the coronary artery could be reversed by exogenous 100 microM L-arginine but not by 100 microM D-arginine. These experiments indicate that Pseudomonas endotoxin induces synthesis of nitric oxide from L-arginine by the vascular endothelium. LPS-mediated production of EDNO by the endothelium, possibly through the action of constitutive nitric oxide synthase (NOSc), may decrease systemic vascular resistance and may be the mechanism of early hypotension characteristic of Pseudomonas septicemia. PMID- 9876304 TI - Histopathological analysis of rat mesentery as a method for evaluating neutrophil migration: differential effects of dexamethasone and pertussis toxin. AB - In the present study, histopathological analysis of rat mesentery was used to quantify the effect of two anti-inflammatory agents, dexamethasone (Dex) and pertussis toxin (Ptx), on leukocyte migration. The intravenous injection of Dex (1 mg/kg) and Ptx (1,200 ng) 1 h prior to the intraperitoneal injection of the inflammatory stimuli lipopolysaccharide (LPS) or formyl-methionyl-leucyl phenylalanine (fMLP) significantly reduced the neutrophil diapedesis (LPS: Ptx = 0.86 +/- 0.19 and Dex = 0.35 +/- 0.13 vs saline (S) = 2.85 +/- 0.59; fMLP: Ptx = 0.43 +/- 0.09 and Dex 0.01 +/- 0.01 vs S = 1.08 +/- 0.15 neutrophil diapedesis/field) and infiltration (LPS: Ptx = 6.29 +/- 1.4 and Dex = 3.06 +/- 0.76 vs S = 15.94 +/- 3.97; fMLP: Ptx = 3.85 +/- 0.56 and Dex = 0.40 +/- 0.16 vs S = 7.15 +/- 1.17 neutrophils/field) induced by the two agonists in the rat mesentery. The inhibitory effect of Dex and Ptx was clearly visible in the fields nearest the venule (up to 200 microns), demonstrating that these anti inflammatory agents act preferentially in the transmigration of neutrophils from the vascular lumen into the interstitial space, but not in cell movement in response to a haptotactic gradient. The mesentery of rats pretreated with Dex showed a decreased number of neutrophils within the venules (LPS: Dex = 1.50 +/- 0.38 vs S = 4.20 +/- 1.01; fMLP: Dex = 0.25 +/- 0.11 vs S = 2.20 +/- 0.34 neutrophils in the lumen/field), suggesting that this inhibitor may be acting at a step that precedes neutrophil arrival in the inflamed tissue. In contrast to that observed with Dex treatment, the number of neutrophils found in mesenteric venules was significantly elevated in animals pretreated with Ptx (LPS: Ptx = 9.85 +/- 2.25 vs S = 4.20 +/- 1.01; fMLP: Ptx = 4.66 +/- 1.24 vs S = 2.20 +/- 0.34 neutrophils in the lumen/field). This discrepancy shows that Ptx and Dex act via different mechanisms and suggests that Ptx prevents locomotion of neutrophils from the vascular lumen to the interstitial space. In conclusion, the method described here is useful for quantifying the inflammatory and anti-inflammatory effect of different substances. The advantage of this histopathological approach is that it provides additional information about the steps involved in leucocyte migration. PMID- 9876306 TI - Partial sequence and toxic effects of granulitoxin, a neurotoxic peptide from the sea anemone Bundosoma granulifera. AB - A neurotoxic peptide, granulitoxin (GRX), was isolated from the sea anemone Bunodosoma granulifera. The N-terminal amino acid sequence of GRX is AKTGILDSDGPTVAGNSLSGT and its molecular mass is 4958 Da by electrospray mass spectrometry. This sequence presents a partial degree of homology with other toxins from sea anemones such as Bunodosoma caissarum, Anthopleura fuscoviridis and Anemonia sulcata. However, important differences were found: the first six amino acids of the sequence are different, Arg-14 was replaced by Ala and no cysteine residues were present in the partial sequence, while two cysteine residues were present in the first 21 amino acids of the other toxins described above. Purified GRX injected i.p. (800 micrograms/kg) into mice produced severe neurotoxic effects such as circular movements, aggressive behavior, dyspnea, tonic-clonic convulsion and death. The 2-h LD50 of GRX was 400 +/- 83 micrograms/kg. PMID- 9876307 TI - Respiratory effects of kynurenic acid microinjected into the ventromedullary surface of the rat. AB - Several studies demonstrate that, within the ventral medullary surface (VMS), excitatory amino acids are necessary components of the neural circuits involved in the tonic and reflex control of respiration and circulation. In the present study we investigated the cardiorespiratory effects of unilateral microinjections of the broad spectrum glutamate antagonist kynurenic acid (2 nmol/200 nl) along the VMS of urethane-anesthetized rats. Within the VMS only one region was responsive to this drug. This area includes most of the intermediate respiratory area, partially overlapping the rostral ventrolateral medulla (IA/RVL). When microinjected into the IA/RVL, kynurenic acid produced a respiratory depression, without changes in mean arterial pressure or heart rate. The respiratory depression observed was characterized by a decrease in ventilation, tidal volume and mean inspiratory flow and an increase in respiratory frequency. Therefore, the observed respiratory depression was entirely due to a reduction in the inspiratory drive. Microinjections of vehicle (200 nl of saline) into this area produced no significant changes in breathing pattern, blood pressure or heart rate. Respiratory depression in response to the blockade of glutamatergic receptors inside the rostral VMS suggests that neurons at this site have an endogenous glutamatergic input controlling the respiratory cycle duration and the inspiratory drive transmission. PMID- 9876308 TI - Effect of Walker 256 tumor growth on intestinal absorption of leucine, methionine and glucose in newly weaned and mature rats. AB - In tumor-bearing rats, most of the serum amino acids are used for synthesis and oxidation processes by the neoplastic tissue. In the present study, the effect of Walker 256 carcinoma growth on the intestinal absorption of leucine, methionine and glucose was investigated in newly weaned and mature rats. Food intake and carcass weight were decreased in newly weaned (NT) and mature (MT) rats bearing Walker 256 tumor in comparison with control animals (NC and MC). The tumor/carcass weight ratio was higher in NT than in MT rats, whereas nitrogen balance was significantly decreased in both as compared to control animals. Glucose absorption was significantly reduced in MT rats (MT = 47.3 +/- 4.9 vs MC = 99.8 +/- 5.3 nmol min-1 cm-1, Kruskal-Wallis test, P < 0.05) but this fact did not hamper the evolution of cancer. There was a significant increase in methionine absorption in both groups (NT = 4.2 +/- 0.3 and MT = 2.0 +/- 0.1 vs NC = 3.7 +/- 0.1 and MC = 1.2 +/- 0.2 nmol min-1 cm-1, Kruskal-Wallis test, P < 0.05), whereas leucine absorption was increased only in young tumor-bearing rats (NT = 8.6 +/- 0.2 vs NC = 7.7 +/- 0.4 nmol min-1 cm-1, Kruskal-Wallis test, P < 0.05), suggesting that these metabolites are being used for synthesis and oxidation processes by the neoplastic cells, which might ensure their rapid proliferation especially in NT rats. PMID- 9876309 TI - The hyperglycemia induced by angiotensin II in rats is mediated by AT1 receptors. AB - We have shown that the renin-angiotensin system (RAS) is involved in glucose homeostasis during acute hemorrhage. Since almost all of the physiological actions described for angiotensin II were mediated by AT1 receptors, the present experiments were designed to determine the participation of AT1 receptors in the hyperglycemic action of angiotensin II in freely moving rats. The animals were divided into two experimental groups: 1) animals submitted to intravenous administration of angiotensin II (0.96 nmol/100 g body weight) which caused a rapid increase in plasma glucose reaching the highest values at 5 min after the injection (33% of the initial values, P < 0.01), and 2) animals submitted to intravenous administration of DuP-753 (losartan), a non-peptide antagonist of angiotensin II with AT1-receptor type specificity (1.63 mumol/100 g body weight as a bolus, i.v., plus a 30-min infusion of 0.018 mumol 100 g body weight-1 min-1 before the injection of angiotensin II), which completely blocked the hyperglycemic response to angiotensin II (P < 0.01). This inhibitory effect on glycemia was already demonstrable 5 min (8.9 +/- 0.28 mM, angiotensin II, N = 9 vs 6.4 +/- 0.22 mM, losartan plus angiotensin II, N = 11) after angiotensin II injection and persisted throughout the 30-min experiment. Controls were treated with the same volume of saline solution (0.15 M NaCl). These data demonstrate that the angiotensin II receptors involved in the direct and indirect hyperglycemic actions of angiotensin II are mainly of the AT1-type. PMID- 9876310 TI - The left ventricular contractility of the rat heart is modulated by changes in flow and alpha 1-adrenoceptor stimulation. AB - Myocardial contractility depends on several mechanisms such as coronary perfusion pressure (CPP) and flow as well as on alpha 1-adrenoceptor stimulation. Both effects occur during the sympathetic stimulation mediated by norepinephrine. Norepinephrine increases force development in the heart and produces vasoconstriction increasing arterial pressure and, in turn, CPP. The contribution of each of these factors to the increase in myocardial performance needs to be clarified. Thus, in the present study we used two protocols: in the first we measured mean arterial pressure, left ventricular pressure and rate of rise of left ventricular pressure development in anesthetized rats (N = 10) submitted to phenylephrine (PE) stimulation before and after propranolol plus atropine treatment. These observations showed that in vivo alpha 1-adrenergic stimulation increases left ventricular developed pressure (P < 0.05) together with arterial blood pressure (P < 0.05). In the second protocol, we measured left ventricular isovolumic systolic pressure (ISP) and CPP in Langendorff constant flow-perfused hearts. The hearts (N = 7) were perfused with increasing flow rates under control conditions and PE or PE + nitroprusside (NP). Both CPP and ISP increased (P < 0.01) as a function of flow. CPP changes were not affected by drug treatment but ISP increased (P < 0.01). The largest ISP increase was obtained with PE + NP treatment (P < 0.01). The results suggest that both mechanisms, i.e., direct stimulation of myocardial alpha 1-adrenoceptors and increased flow, increased cardiac performance acting simultaneously and synergistically. PMID- 9876311 TI - What a lousy outlook. PMID- 9876312 TI - Squeezing tubes: a case of remodeling and regulation: coronary reserve in hypertensive heart disease. PMID- 9876313 TI - Role of protein kinase C as a cellular mediator of ischemic preconditioning: a critical review. AB - Protein kinase C (PKC) has been proposed as a primary cellular mediator of ischemic preconditioning. However, the role of PKC in eliciting cardioprotection remains controversial. In this review, we summarize the evidence for and against the 'PKC hypothesis' of preconditioning, discuss the important technical limitations currently hampering PKC research, and suggest new approaches (i.e. PKC isoform-specific biochemical assays combined with immunoblotting techniques) that might aid in the definite resolution of this issue. PMID- 9876314 TI - Role of cardiac chloride currents in changes in action potential characteristics and arrhythmias. AB - Various types of Cl- currents have been recorded in cardiac myocytes from different regions of the heart and in different species. With few exceptions, most of these currents are not active under basal conditions, but are activated under the influence of various agonists and by physical stress. These channels are distributed nonuniformly, depending on the cell type, tissue and region of the heart. Therefore, Cl- current activation may influence membrane potential and impulse formation differently in different cells, and may play a role in arrhythmogenesis. Among these Cl- currents, the protein kinase A-activated Cl- current (I Cl.PKA), the stretch- or swelling-activated Cl- current (I Cl.SWELL) and the Ca(2+)-activated Cl current (I Cl.Ca) comprise the major anion currents that modify cardiac electrical activity. These currents exhibit outward-going rectification, or are predominantly activated at depolarized voltages and, thus, contribute significantly to shortening of the action potential duration but little to diastolic depolarization. The action potential shortening by Cl- current activation may not only perpetuate reentry by shortening the refractory period in a reentry pathway, but may also prevent the development of early afterdepolarization and triggered activity caused by the prolongation of action potentials. I Cl.Ca contributes to delayed afterdepolarization at diastolic potentials in Ca(2+)-overloaded cells. Another factor limiting the influence of Cl- currents on diastolic potentials is the presence of a predominantly opposing background K+ current, except at the nodal regions that lack these K+ channels, or under conditions of decreased K+ conductance. Therefore, the contribution of Cl- currents to the genesis of arrhythmias may depend on their association with the conductance of other ions, especially that of K+. PMID- 9876315 TI - Cardiovascular steroid actions: swift swallows or sluggish snails? AB - Steroid actions on the vascular wall have been thought to depend on direct, genomic mechanisms being characterized by a considerable delay and on secondary events, including changes of coagulation, plasma lipids, and renal electrolyte and volume regulation. Recently, rapid effects of steroids on the vascular wall have been reported being clearly incompatible with the classical theory of genomic steroid action. As these effects occur in classical target tissues for genomic steroid action, and modulation of intracellular signaling has been shown to influence genomic steroid action, a two-step model of steroid action was developed integrating both genomic and nongenomic aspects and their possible interaction. This review summarizes recent studies on both types of direct, vascular steroid actions, the swift and the sluggish ones, and discusses the role of these actions in regulation of circulatory homeostasis and their potential therapeutic implications. PMID- 9876316 TI - Vagal nerve stimulation releases vasoactive intestinal peptide which significantly increases coronary artery blood flow. AB - OBJECTIVE: To determine the effects of vasoactive intestinal peptide (VIP), released endogenously from cardiac vagal nerves, on coronary artery blood flow (CBF). METHODS: We determined the effects of vagal nerve stimulation (VNS) at frequencies of 10, 15, 20, and 30 Hz on left circumflex coronary artery (LCx) blood flow. The increases in CBF during VNS were compared with the increases in CBF produced by exogenous VIP and also nitroglycerin (NTG). In 18 anesthetized open chest mongrel dogs, we blocked the muscarinic and beta-adrenergic receptors with atropine and propranolol. We controlled heart rate and aortic pressure by right atrial pacing and an arterial reservoir. CBF was measured in the LCx with a Doppler flow probe. A 25 gauge catheter was placed in the proximal LCx to inject the VIP receptor antagonist [4Cl-D-Phe6Leu17]VIP, VIP, NTG, or vehicle. CBF, aortic and ventricular pressures, ventricular contractility (+dp/dt(max)) and relaxation (-dp/dt(min)) and the EKG were measured. RESULTS: VNS (0.5 ms, 20 V, 5 min.) at 20 Hz maximally increased CBF by 62 +/- 14% at 5 min from 71 +/- 10 to 115 +/- 19 ml/min (p < 0.01). VNS at 10, 15, and 30 Hz increased CBF by 6 +/- 1%, 24 +/- 5%, and 24 +/- 7%, respectively (all p < 0.05 vs control). Following 20 Hz VNS, CBF returned toward the baseline over 30 min. Aortic and left ventricular (LV) pressures, LV +dp/dt(max) and LV-dp/dt(min) did not significantly change. After the direct administration of [4Cl-D-Phe6Leu17]VIP into the LCx, VNS increased CBF by only 10 +/- 4% (p = NS). Exogenous VIP, in doses of 9.0 x 10( 11) to 2.1 x 10(-9) mol, increased CBF by 106 +/- 17% to 169 +/- 17% (all p < 0.01 vs control). NTG, in doses of 2.2 x 10(-8) to 1.7 x 10(-7) mol, increased CBF by 101 +/- 15% to 169 +/- 20% (all p < 0.01 vs control). These increases in CBF persisted during the 1 to 2 min injection period and returned to the baseline within 5 min. Neither VIP nor NTG significantly changed the heart rate, aortic or LV pressures, LV +dp/dt(max) or LV -dp/dt(min). VNS at 20 Hz, exogenous VIP, 9.0 x 10(-11) mol, and exogenous NTG, 2.2 x 10(-8) to 4.4 x 10(-8) mol, produced equivalent increases in CBF by analysis of variance determination. CONCLUSION: The present experiments suggest that VNS releases VIP which directly dilates coronary arteries and significantly increases coronary artery blood flow. PMID- 9876317 TI - Antifibrillatory efficacy of ersentilide, a novel beta-adrenergic and Ikr blocker, in conscious dogs with a healed myocardial infarction. AB - OBJECTIVES: IKr blockade is ineffective in preventing ventricular fibrillation elicited by the interaction between acute myocardial ischemia and elevated sympathetic activity. This depends in part on the fact that adrenergic activation offsets more than 50% of the action potential prolonging effect of IKr blockade, and thus impairs its primary mechanism of action. This study examined the antifibrillatory effect of ersentilide (CK-3579), a novel antiarrhythmic agent which combines blockade of the rapid component of the delayed rectifier potassium channel (IKr) with relatively weak beta-adrenergic blockade, in a conscious canine model of lethal arrhythmias. METHODS: Ersentilide was tested in 19 dogs with a healed myocardial infarction (MI) undergoing two minutes of circumflex artery occlusion (CAO) during sub-maximal treadmill exercise. Epicardial monophasic action potential duration was measured before and after ersentilide in 8 anesthetized open chest dogs at baseline and during stimulation of the left stellate ganglion at constant paced heart rate. RESULTS: In the control tests 13 of the 19 dogs had ventricular fibrillation (VF) during the exercise and ischemia test, 6 did not. During a subsequent exercise test, ersentilide prevented VF in 85% (11 of 13) of the high risk animals and showed no proarrhythmic effects in the 6 dogs without arrhythmias in the initial test. Ersentilide lowered heart rate at all levels of exercise and during acute myocardial ischemia. The antifibrillatory effect was maintained in 3 of 4 dogs in which heart rate was kept at control levels by atrial pacing. Ersentilide prolonged left ventricular monophasic action potential duration by 30% (from 179 +/- 6 ms to 233 +/- 5 ms, p < 0.001) at a 360 ms cycle length and completely prevented its shortening during sympathetic stimulation. CONCLUSIONS: The combination of IKr and weak beta adrenergic blockade, using ersentilide, represents a very effective and safe antiarrhythmic intervention able to overcome the limitations present in drugs devoid of any antiadrenergic effect. Such a combination may be very useful in the management of post-myocardial infarction patients at high arrhythmic risk. PMID- 9876318 TI - Growth hormone preserves cardiac sarcoplasmic reticulum Ca2+ release channels (ryanodine receptors) and enhances cardiac function in cardiomyopathic hamsters. AB - OBJECTIVE: Growth hormone (GH) improves cardiac function in experimental models of heart failure and human dilated cardiomyopathy. However, the mechanism by which GH increases myocardial contractility is not entirely clear. Our aim was to examine the effects of GH on cardiac function and cardiac sarcoplasmic reticulum Ca2+ release channels (ryanodine receptors, RyR) in the hearts of UM-X7.1 cardiomyopathic hamsters during the development of heart failure. METHODS: Experimental and healthy control hamsters were examined at the age of 20 weeks. Recombinant human GH (2 mg/kg/day, s.c.) or vehicle was then administered for 3 weeks. We examined (i) the in vivo left ventricular (LV) size and LV systolic function using transthoracic echocardiography, (ii) the density (Bmax) and affinity (Kd) of high-affinity [3H] ryanodine binding sites in crude homogenates from normal and cardiomyopathic hamster hearts. RESULTS: Vehicle-treated UM-X7.1 hamsters exhibited significant increases in left ventricular end-diastolic diameter and end-systolic diameter (LVESd), and a significant decrease in LV fractional shortening (FS). GH-treatment attenuated the increase in LVESd and reduced the LV chamber size, and also significantly increased LVFS. Vehicle treated UM-X7.1 hamsters exhibited a significantly lower Bmax than control hamsters (0.34 +/- 0.04 vs 0.44 +/- 0.06 pmol/mg, p < 0.05), and the treatment with GH in UM-X7.1 hamsters significantly attenuated the reduction of Bmax (0.42 +/- 0.03 pmol/mg vs vehicle-treated group (0.34 +/- 0.04 pmol/mg), p < 0.05). Kd did not differ significantly between the experimental groups. In normal control hamsters, GH treatment with this dose did not significantly enhance LV systolic function or the density of RyRs. There was no significant difference in terms of the connective-tissue volume-fraction, myocyte size and capillary density between the GH- and vehicle-treated groups of UM-X7.1 hamsters. CONCLUSIONS: GH treatment may improve cardiac function by preserving the density of RyRs and enhancing cellular function in cardiomyopathic hamster hearts. PMID- 9876319 TI - Extracellular adenosine levels and cellular energy metabolism in ischemically preconditioned rat heart. AB - OBJECTIVE: Microdialysis and 31P-NMR spectroscopy were used to test opposing hypotheses that ischemic preconditioning inhibits adenine nucleotide degradation and purine efflux, or that preconditioning activates cardiovascular adenosine formation to provide enhanced cardioprotection. METHODS: 31P-NMR spectra and matching interstitial fluid (ISF) or venous effluent samples were obtained from Langendorff perfused rat hearts. Control hearts (n = 9) underwent 30 min of global normothermic ischemia and 30 min reperfusion. Preconditioned hearts (n = 6) were subjected to a 5 min ischemic episode and 10 min reflow prior to 30 min ischemia and 30 min reperfusion. Effects of repetitive ischemia-reperfusion (3 x 5 min ischemic episodes) on adenosine levels and energy metabolism were also assessed (n = 8). RESULTS: Preconditioning improved post-ischemic recovery of heart rate x left ventricular developed pressure (71 +/- 5 vs 43 +/- 8%, P < 0.05) and end-diastolic pressure (14 +/- 3 vs 29 +/- 4 mmHg, P < 0.05) compared with control hearts, respectively. Preconditioning did not alter intracellular ATP, phosphocreatine (PCr), inorganic phosphate (Pi), H+ or free Mg2+ during global ischemia, but improved recoveries of PCr, Pi, and delta GATP on reperfusion. ISF adenosine increased more than 20-fold during 30 min ischemia. The 5 min preconditioning episode increased ISF adenosine 3-fold, and reduced ISF adenosine and inosine during subsequent prolonged ischemia by up to 75%. Venous purine levels during reperfusion were also reduced by preconditioning. Accumulation of adenosine in ISF and venous effluent during repetitive ischemia was progressively reduced despite comparable changes in substrate for adenosine formation via 5'-nucleotidase, (5'-AMP), and in allosteric modulators of this enzyme (Mg2+, H+, Pi, ADP, ATP). CONCLUSIONS: (i) Ischemic preconditioning reduces interstitial and vascular adenosine levels during ischemia-reperfusion, (ii) reduced ISF adenosine during ischemia is not due to reduced ischemic depletion of adenine nucleotides in preconditioned rat hearts, (iii) preconditioning may inhibit adenosine formation via 5'-nucleotidase in ischemic rat hearts, and (iv) improved functional recovery with preconditioning is unrelated to metabolic/bioenergetic changes during the ischemic insult, but may be related to improved post-ischemic recovery of [Pi] and delta GATP in this model. PMID- 9876320 TI - Left coronary pressure-flow relations of the beating and arrested rabbit heart at different ventricular volumes. AB - OBJECTIVE: To study the effect of cardiac contraction on left coronary artery pressure-flow relations at different vascular volumes and to compare these relations in the beating heart with those in the heart arrested in systole and diastole. METHODS: Maximally vasodilated, Tyrode perfused, rabbit hearts (n = 6) with an intra-ventricular balloon were used. The left coronary artery was separately perfused via a cannula in the left main coronary artery. The slopes and the intercepts of left coronary pressure-flow relations were determined in the beating and arrested heart at different chamber volumes. A 3-factor design with repeated measures was used to compare the effect of three factors: phase of contraction (systole and diastole), chamber volume (V0 and V1, left ventricular end-diastolic pressure 1.4 and 20 mm Hg, respectively) and the type of contraction (beating and arrested; a measure of capacitive effects). RESULTS: The phase of contraction has a significant effect on the intercepts (> 40 mmHg, p = 0.00032) but not on the slopes of the pressure-flow relations. Chamber volume had a small effect on the intercepts (< 5 mm Hg, p = 0.037), but not on the slopes of the pressure-flow relations. The type of contraction has a significant effect on the slopes (approximately 10%, p = 0.00021) but not on the intercepts of the pressure-flow relations. CONCLUSIONS: In the isolated Tyrode perfused rabbit heart left coronary pressure-flow relations are mainly determined by contraction, while left ventricular chamber volume and capacitive effects contribute little. PMID- 9876321 TI - Insulin signalling in heart involves insulin receptor substrates-1 and -2, activation of phosphatidylinositol 3-kinase and the JAK 2-growth related pathway. AB - OBJECTIVE: Hyperinsulinemia is a common feature of obesity and hypertension and may be associated with abnormal metabolism and growth of heart muscle and vascular wall. Most of the known actions of insulin were characterised in muscle, adipose tissue and liver. In this study we investigate the initial steps of insulin signalling in rat heart. METHODS: After insulin infusion in the cava vein of male Wistar rats, the insulin receptor, insulin receptor substrates-1 and -2, phosphatidylinositol 3-kinase activity and Janus kinase (JAK) 2 engagement were studied by immunoprecipitation and immunoblot of heart extracts. RESULTS: An insulin load induces rapid autophosphorylation of the insulin receptor which is followed by the phosphorylation of insulin receptor substrates-1 and -2. The phosphorylation of these early intracellular substrates leads to the association of the p85 subunit of phosphatidylinositol 3-kinase and subsequent activation of its catalytic p110 subunit. Besides activation of the lipid metabolising enzyme phosphatidylinositol 3-kinase, the phosphorylation of insulin receptor substrates 1 and -2 engages the intracellular kinase JAK 2 and induces JAK 2-STAT 1 complex formation. CONCLUSIONS: We demonstrate that the early steps of insulin signalling in heart include the phosphorylation-activation of the insulin receptor, engagement of insulin receptor substrates-1 and -2 with the consequent activation of phosphatidylinositol 3-kinase and the involvement of the recently discovered growth related pathway--JAK 2-STAT 1. PMID- 9876322 TI - Passive transmission of ischemic ST segment changes in low electrical resistance myocardial infarct scar in the pig. AB - OBJECTIVES: To analyze the passive electrical properties of a healed infarction and assess their role on transmission of contiguous ischemic ST segment potential changes. METHODS: We measured tissue resistivity (omega cm) at 1 kHz and the epicardial ST segment during 1 h of proximal reocclusion of the left anterior descending (LAD) coronary artery in 12 anesthetized pigs with one-month-old transmural infarction elicited by LAD ligature below the first branch. The impedance spectrum (1 to 1000 kHz) of normal and infarcted myocardium was measured in seven other pigs with similar infarctions. Electrical transmission of current pulses (30 microA) in infarcted tissue and in test solutions was also investigated. RESULTS: The infarct scar has a lower than normal resistivity (110 +/- 30 omega cm vs. 235 +/- 60 omega cm, p < 0.0001) and, unlike the normal myocardium, resistivity and phase angle of the scar did not change at increasing current frequencies, reflecting no capacitative response. LAD reocclusion induced a resistivity rise (510 +/- 135 omega cm, p < 0.01) and a ST segment elevation (0.6 +/- 0.7 to 9.5 +/- 5.1 mV, p = 0.002) in the ischemic peri-infarction zone, whereas the infarcted area showed ST segment elevation (0.5 +/- 0.5 to 3.8 +/- 2.6 mV, p = 0.03) with no resistivity changes. Potential decay of both ST segment and current pulses in the scar and in 0.9% NaCl solution was less than 1 mV/mm. Transmural deposition of connective tissue was seen in the center of the infarction. CONCLUSIONS: A one-month-old transmural infarction is a low resistance, noncapacitative medium that allows a good transmission of current pulses and of ST segment potential changes generated by contiguous peri infarction ischemia. PMID- 9876323 TI - Intra-coronary administration of L-arginine aggravates myocardial stunning through production of peroxynitrite in dogs. AB - OBJECTIVE: The aim of this study was to investigate how the enhanced nitric oxide (NO) production by intra-coronary infusion of L-arginine acts in myocardial stunning in dogs by focusing on the involvement of peroxynitrite, a reaction product of NO and superoxide anion. METHODS AND RESULTS: Dogs were divided into six groups; a control non-treated group (CON, n = 9), and NG-nitro L-arginine methyl ester (L-NAME, n = 6), 1 mM L-arginine (L-ARG, n = 8), D-arginine (D-ARG, n = 6), L-arginine plus superoxide dismutase (L-ARG + SOD, n = 6), and SOD alone (SOD, n = 6) treated groups. L-NAME, or L- or D-arginine was continuously infused into the left anterior descending coronary artery (LAD) starting just prior to reperfusion, whereas SOD was intravenously injected before occlusion. During 120 min of reperfusion after 15 min occlusion of LAD, myocardial contractile function in the ischemic region gradually recovered and reached approximately 70% of the preischemic level in CON, D-ARG and SOD, but it remained dyskinetic (-46%) in L ARG. On the other hand, it was improved in L-NAME (90%). Tissue malondialdehyde was elevated (p < 0.005) after reperfusion, and myocardial NO metabolites measured by an intratissue-microdialyzer increased (approximately 150%, p < 0.05) in the ischemic region during reperfusion in L-ARG but not in the CON, L-NAME, D ARG or SOD groups. In the L-ARG + SOD group, L-arginine-induced contractile dysfunction and elevation of malondialdehyde were prevented, but the increase in NO metabolites remained. These results suggest that L-arginine aggravated myocardial stunning through oxidative stress and the cytotoxicity was caused by NO derivatives but not by NO itself. The formation of nitrotyrosine, a footprint of peroxynitrite, was immunohistochemically confirmed in the ischemic region of L ARG. CONCLUSIONS: Our results demonstrate for the first time in vivo that NO has a detrimental role in myocardial stunning through the production of peroxynitrite. PMID- 9876324 TI - Heat stress-induced resistance to myocardial infarction in the isolated heart from transgenic [(mREN-2)27] hypertensive rats. AB - OBJECTIVE: Heat stress (HS) is known to confer protection against ischaemia reperfusion injury, including mechanical dysfunction and myocardial necrosis. However, the effects of disease states on this HS-induced cytoprotective response are less known. Therefore, we investigated the effects of prior heat stress on the infarct size in the isolated rat heart and on the myocardial heat stress protein (HSP) 72 synthesis, in transgenic [(mREN-2)27] hypertensive (TGH) rats or normotensive (NT) controls. METHODS: TGH or NT rats were either heat stressed (42 degrees C for 15 min) or sham anaesthetised. After 24 h, their hearts were isolated, perfused using the Langengorff technique, and subjected to a 35-min occlusion of the left coronary artery followed by 120 min of reperfusion. Myocardial HSP72 content was measured 24 h after HS or sham treatment using electrophoresis coupled with Western blot analysis. RESULTS: Infarct-to-risk (I/R) ratio was significantly reduced in HS (15.5 +/- 1.2%) compared to sham (42.2 +/- 2.1%) hearts of NT rats. This reduction in infarct size was maintained in TGH hearts (I/R: 20.0 +/- 1.0 vs. 48.0 +/- 3.8%). Risk zones were similar between all experimental groups. The incidence of ventricular arrhythmias during ischaemia and reperfusion periods was not different between the four experimental groups. Western blot analysis of the myocardial HSP72 content showed a heat stress-induced increase of this protein, in both TGH and NT animals. CONCLUSION: These results demonstrate that the myocardial protective effect induced by heat stress could extend to a pathological animal model like the transgenic [(mREN 2)27] hypertensive rat and is correlated with a myocardial HSP72 induction. PMID- 9876325 TI - Observation and basis of improved blood flow to the distal latissimus dorsi muscle: a case for electrical stimulation prior to grafting. AB - OBJECTIVE: Dynamic cardiomyoplasty, using a functional graft of the latissimus dorsi muscle, has shown promise as a treatment for selected patients with advanced heart failure. The success of this approach depends on maintaining the viability of the muscle, whose distal portion is susceptible to ischaemic damage. We investigated the effects of surgical mobilization on regional muscle blood flow and the influence of electrical stimulation of the muscle. METHODS: Ten sheep were randomly assigned to two equal groups. In one group, the latissimus dorsi muscle was stimulated continuously in situ at 2 Hz for two weeks; in the other group, the muscle was not stimulated. Regional blood flows in the muscles were determined by a fluorescent microsphere technique. Serial measurements were made (a) under baseline conditions before intervention, (b) with the thoracodorsal artery occluded and (c) after interruption of the perforating collateral arteries. RESULTS: Surgical mobilization of the unstimulated latissimus dorsi muscles had little effect on blood flow in the proximal region, which remained at 93.1 +/- 16.9% of baseline (mean +/- SEM). The distal region was rendered significantly more ischaemic (55.8 +/- 13.5% of baseline, p < 0.002 compared to the proximal region). Electrical prestimulation abolished any significant proximodistal gradient in blood flow and improved distal muscle perfusion following mobilization (proximal vs. distal: 75.0 +/- 8.8 vs. 63.0 +/- 10.9%; p > 0.4). CONCLUSIONS: Distal muscle ischaemia occurred when the entire latissimus dorsi muscle was acutely elevated on the thoracodorsal pedicle alone. Electrical prestimulation of the muscle in situ improved the thoracodorsal perfusion of the distal muscle by abolishing the proximal-to-distal gradient in flow, with a substantial benefit to distal flow after mobilization. Although electrical stimulation is known to induce vascular proliferation, we argue that this effect of stimulation is brought about mainly by enhancement of the flow through anastomotic connections between proximal and distal arterial territories. PMID- 9876326 TI - Selective activation of adenosine A3 receptors with N6-(3-chlorobenzyl)-5'-N methylcarboxamidoadenosine (CB-MECA) provides cardioprotection via KATP channel activation. AB - OBJECTIVE: The aim of this study was to characterize the adenosine A3 receptor agonist, N6-(3-chlorobenzyl)-5'-N-methylcarboxamidoadenosine (CB-MECA), evaluate its ability to reduce myocardial ischemia/reperfusion injury and determine the role of KATP-channel activation in A3 receptor-mediated cardioprotection. METHODS: Binding affinities and adenylate cyclase inhibition were examined in CHO cells expressing rabbit recombinant adenosine A1 or A3 receptors. Infarct size (normalized for area-at-risk; % IA/AAR) was measured in buffer-perfused rabbit hearts exposed to 30-min regional ischemia and 120 min of reperfusion. RESULTS: CB-MECA was 100-fold selective for A3 vs. A1 receptors (A3 Ki: 1 nM; A1 Ki: 105 nM). Five-min perfusion with CB-MECA before ischemia/reperfusion elicited a concentration-dependent reduction in infarct size (EC50: 0.3 nM). The CB-MECA dependent cardioprotection (control: 58 +/- 2; CB-MECA: 21 +/- 3% IA/AAR) was unchanged by an A1-selective concentration of the antagonist, BWA1433, but was completely prevented (P < 0.05) by a nonselective (A1/A3) concentration (55 +/- 6% IA/AAR). The KATP channel inhibitors, glibenclamide and 5-HD, had no effect on control infarct size, yet significantly (P < 0.05) blunted the CB-MECA-dependent cardioprotection (glibenclamide: 49 +/- 6; 5-HD: 58 +/- 4% IA/AAR). CONCLUSIONS: CB-MECA is a novel 100-fold A3 receptor-selective agonist which should prove useful for elucidating A3-dependent mechanisms in the rabbit heart. Selective stimulation of adenosine A3 receptors with CB-MECA reduces myocardial ischemia/reperfusion injury via a mechanism which involves activation of KATP channels. PMID- 9876327 TI - Beta-adrenergic signal transduction following carvedilol treatment in hypertensive cardiac hypertrophy. AB - OBJECTIVE: Treatment with the beta-blocker carvedilol leads to an improvement of outcome and ejection fraction in heart failure. These effects occur without affecting the number of beta-adrenergic receptors, as determined in right ventricular biopsies from patients with heart failure. This study was aimed at investigating the effects of carvedilol on beta-adrenergic signal transduction alterations in a model of left ventricular pressure overload, which is characterized by sympathetic activation and a desensitized beta-adrenergic signal transduction. METHODS: Transgenic rats with overexpression of renin [TG(mREN2)27] were treated with carvedilol (30 micrograms/kg) or held under control conditions and were compared with Sprague-Dawley rats. Myocardial beta-adrenoceptors (125I labeled iodocyanopindolol binding), Gi alpha (pertussis toxin labeling), Gs alpha activity (reconstitution into cyc--S49 membranes) and adenylyl cyclase activity were measured. Blood pressure and heart rate, increase in heart rate during sacrifice and pressure rate products were determined. RESULTS: beta-Adrenoceptors were downregulated and Gi alpha-protein levels were significantly increased, producing a desensitization of basal, isoprenaline- and guanine nucleotide stimulated adenylyl cyclase activity compared to controls. Carvedilol reduced heart rate, blood pressure and pressure rate product in TG(mREN2)27. Carvedilol did not restore biochemical alterations, but even further reduced beta adrenoceptor numbers and adenylyl cyclase. It exhibited a two affinity state, guanine nucleotide-sensitive binding to cardiac beta-adrenergic receptors similar to isoprenaline but different from metoprolol. CONCLUSIONS: Carvedilol did not restore beta-adrenergic signal transduction at concentrations producing antiadrenergic effects in vivo. This effect might be due to an atypical guanine nucleotide-dependent interaction with beta-adrenergic receptors. Thus, ancillary properties could explain the recently reported beneficial effects in patients with heart failure independent from an upregulation of beta-adrenergic receptors. PMID- 9876328 TI - Early captopril treatment inhibits DNA synthesis in endothelial cells and normalization of maximal coronary flow in infarcted rat hearts. AB - OBJECTIVES: Cardiac remodeling due to myocardial infarction (MI) includes myocyte hypertrophy, collagen deposition, a rise in DNA synthesis, and normalization of initially diminished maximal coronary bloodflow. Previously, we demonstrated that early captopril treatment can prevent the rise in total DNA synthesis, collagen deposition and hypertrophy. In the present experiments, we investigated the effects of captopril or perindoprilat treatment on cardiac endothelial cell proliferation and maximal coronary flow. METHODS: MI was induced by ligation of the left coronary artery in Wistar rats. Sham-operated and infarcted rats were treated with captopril (12 mg/kg.d s.c.) from either day 0-21 (early) or day 21 35 (late) after surgery. In isolated retrogradely perfused rat hearts, maximal coronary flow was determined following maximal dilatation with nitroprusside and adenosine (1 mM each). In separate groups, sections of hearts of sham-operated and MI rats treated with BrdU (day 7-14) and either captopril or perindoprilat (1 mg/kg.d s.c.; day 0-14) were double stained with a monoclonal anti-BrdU antibody and the lectin GSI. The total fraction of DNA synthesizing cells and its proportion of endothelial cells was determined. RESULTS: Maximal coronary flow was completely normalized in MI hearts within three weeks after surgery. Early captopril, but not late captopril, inhibited the normalization of maximal coronary flow in MI hearts (Early: sham, 27.4 +/- 1.0; MI, 21.2 +/- 1.4 ml/min; P < 0.05; mean +/- SEM) without affecting the hypertrophic response. The total fraction of DNA synthesizing cells was significantly increased in MI hearts (sham: 7.6 +/- 1.9; MI: 14.9 +/- 2.2%). The proportion of endothelial cells, however, was comparable in sham-operated and infarcted hearts (sham: 30 +/- 3; MI: 33 +/- 3%). Both early captopril and perindoprilat treatment inhibited total DNA synthesis in MI hearts. Only in captopril pre-treated hearts, this inhibition was associated with a disproportionate inhibition of the endothelial cell proliferation (10.3 +/- 2.0%). CONCLUSION: Early captopril treatment inhibits endothelial cell proliferation and coronary vessel growth following MI, which seems to be partly due to inhibition of the renin angiotensin system. PMID- 9876329 TI - Coenzyme Q10 treatment improves the tolerance of the senescent myocardium to pacing stress in the rat. AB - OBJECTIVE: In elderly patients the results of cardiac interventions are inferior to those in the young. A possible contributing factor is an age-related reduction in cellular energy transduction during the intervention which may induce aerobic or ischemic stress. To investigate whether coenzyme Q10 (CoQ10) improves the response to aerobic stress, functional recoveries of senescent and young rat hearts after rapid pacing were compared with or without CoQ10. METHODS: Young (4.8 +/- 0.1 months) and senescent (35.3 +/- 0.2 months) rats were given daily intraperitoneal injections of CoQ10 (4 mg/kg) or vehicle for 6 weeks. Their isolated hearts were rapidly paced at 510 beats per minute for 120 min to induce aerobic stress without ischemia. RESULTS: In senescent hearts pre-pacing cardiac work was 74% and oxygen consumption (MVO2) 66% of that in young hearts. CoQ10 treatment abolished these differences. After pacing, the untreated senescent hearts, compared to young, showed reduced recovery of pre-pacing work, (16.8 +/- 4.3 vs. 44.5 +/- 7.4%; P < 0.01). CoQ10 treatment in senescent hearts improved recovery of work, (48.1 +/- 4.1 vs. 16.8 +/- 4.3%; P < 0.0001) and MVO2 (82.1 +/- 2.8 vs. 61.3 +/- 4.0%; P < 0.01) in treated versus untreated hearts respectively. Post-pacing levels of these parameters in CoQ10 treated senescent hearts were as high as in young hearts. CONCLUSIONS: (1) Senescent rat hearts have reduced baseline function and reduced tolerance to aerobic stress compared to young hearts. (2) Pre-treatment with CoQ10 improves baseline function of the senescent myocardium and its tolerance to aerobic stress. PMID- 9876332 TI - The cell adhesion molecule E-cadherin is widely expressed in human atherosclerotic lesions. AB - OBJECTIVE: Various cell adhesion molecules are expressed in atherogenesis and the significance of their involvement in atherosclerotic lesion formation is well appreciated. In the present work, we examined whether the Ca(2+)-dependent cell adhesion molecule E-cadherin is also involved in atherogenesis. METHODS: Specimens of carotid artery and aorta were obtained at operation. Expression of E cadherin was studied by an immunohistochemical method. The nature of E-cadherin expressing cells was examined by comparative analysis of consecutive sections and by a double immunostaining procedure. An immunohistochemical approach was also applied to examine how the accumulation of oxidised low density lipoproteins (LDL) by intimal cells is associated with E-cadherin expression. RESULTS: No E cadherin+ cells were found in normal non-atherosclerotic intima but E-cadherin+ cells were present in 96% of the atherosclerotic lesions. In atherosclerotic intima, E-cadherin was expressed by intimal cells showing varying degrees of transformation into foam cells. These E-cadherin+ cells also contained oxidised LDL in their cytoplasm. Differing numbers of CD68+ foam cells (15% to 60%) expressed E-cadherin but all the CD68+ macrophages without signs of transformation into foam cells were negative for E-cadherin. Neither smooth muscle cells nor foam cells of smooth muscle cell origin (smooth muscle alpha actin+) were found to be positive for E-cadherin. T-cells (CD3+) and endothelial cells (von Willebrand factor+) were also negative for E-cadherin. Only a few vascular dendritic cells (S-100+) expressed E-cadherin and their expression was weak. We also found that a large proportion (40% to 85%) of E-cadherin+ cells did not stain with any cell-type specific markers. CONCLUSIONS: The finding that E cadherin is expressed in atherosclerotic lesions expands our knowledge of cell adhesion molecules involved in atherogenesis. That E-cadherin is expressed in intimal cells transforming into foam cells suggests that lipid accumulation might be associated with the alteration and reorganisation of cell-to-cell interactions in atherogenesis. The present observations might assist in understanding the mechanisms associated with intracellular lipid accumulation. PMID- 9876331 TI - Phospholipid source and molecular species composition of 1,2-diacylglycerol in agonist-stimulated rat cardiomyocytes. AB - OBJECTIVE: The aim was to investigate the consequences of simultaneous stimulation of phospholipase C and D by agonists for the molecular species composition of 1,2-diacylglycerol and phospholipids in cardiomyocytes. METHODS: Serum-free cultured neonatal rat cardiomyocytes were stimulated by endothelin-1, phenylephrine or phorbolester. The molecular species of 1,2-diacylglycerol (in mol%) and those derived from phosphatidylcholine and phosphatidylinositol were analyzed by high-performance liquid chromatography and their absolute total concentration (nmol per dish) by gas-liquid chromatography. Phospholipids were labelled with [14C]glycerol or double-labelled with [14C]16:0 and [3H]20:4n6 for measurements of respectively, the amount of or relative rate of label incorporation into 1,2-diacylglycerol. RESULTS: The major molecular species of 1,2-diacylglycerol in unstimulated cells was found to be 18:0/20:4 (57 mol%). The same species was observed predominantly in phosphatidylinositol (73 mol% compared to 11 mol% in phosphatidylcholine). A significant decrease (about 10 mol%) was found for the 18:0/20:4 species of 1,2-diacylglycerol during stimulation (10-40 min) with endothelin-1 or phorbolester, but not phenylephrine. The results of the double-labelling experiments were consistent with the latter finding: the ratio [3H]20:4 over [14C]16:0 in 1,2-diacylglycerol decreased from 1.70 in the control to 1.40 during 10-min endothelin-1 or phorbolester stimulation, but not during phenylephrine stimulation. The [14C]glycerol incorporation into 1,2 diacylglycerol remained relatively constant under agonist-stimulated conditions as did the total concentration of 1,2-diacylglycerol. CONCLUSIONS: 1,2 Diacylglycerol present in unstimulated cardiomyocytes is likely derived from phosphatidylinositol. During stimulation with endothelin-1 and phorbolester, but not phenylephrine, phosphatidylcholine becomes an increasingly important source for 1,2-diacylglycerol due to sustained activation of phospholipase D. The 1,2 diacylglycerol level remains relatively constant during agonist stimulation which strongly indicates that particular molecular species of 1,2-diacylglycerol more than its total concentration determine the activation of protein kinase C isoenzymes. PMID- 9876330 TI - Role of phosphatidylinositol 3-kinase activation in the hypertrophic growth of adult ventricular cardiomyocytes. AB - OBJECTIVE: The present study investigated whether activation of phosphatidylinositol 3-kinase (PI3-kinase) is involved in the stimulation of hypertrophic growth of adult ventricular cardiomyocytes under alpha- or beta adrenoceptor stimulation. METHODS: Adult ventricular rat cardiomyocytes were used either directly after isolation (day 1 culture) or after cultivation for 6 days in presence of 20% fetal calf serum (day 7 culture). PI3-kinase activity was determined in extracts of cardiomyocytes after immunoprecipitation with an antibody against the p85 subunit of PI3-kinase. The influence of PI3-kinase inhibition on myocardial growth was determined using the specific PI3-kinase inhibitors wortmannin and LY294002. RESULTS: In day 1 cultures alpha-adrenoceptor stimulation, but not beta-adrenoceptor stimulation caused activation of PI3 kinase. In response to alpha-adrenoceptor stimulation but not beta-adrenoceptor stimulation an acceleration of protein synthesis (incorporation of 14C phenylalanine) and an increase in the total masses of cellular protein and RNA was observed. In these cultures inhibition of PI3-kinase attenuated the acceleration of protein synthesis and the increase in cellular masses of protein or RNA in response to alpha-adrenoceptor stimulation. In day 7 cultures alpha- and beta-adrenoceptor stimulation caused activation of PI3-kinase and increased protein synthesis. In these cultures inhibition of PI3-kinase attenuated the growth response to alpha- and beta-adrenoceptor stimulation. CONCLUSIONS: PI3 kinase activation via protein kinase C-dependent or cAMP-dependent pathways is required for hypertrophic growth of adult cardiomyocytes. PMID- 9876333 TI - Upregulation of angiotensin converting enzyme by atrial natriuretic peptide and cyclic GMP in human endothelial cells. AB - OBJECTIVE: To examine the role of atrial natriuretic peptide (ANP) and cyclic GMP in the regulation of angiotensin converting enzyme (ACE) in cultured human endothelial cells. METHODS: Cultured endothelial cells from human umbilical veins (HUVEC) were treated with ANP (0.3-30 nM), 8-Br-cGMP (1-100 microM), Rp-8-Br-PET cGMPS (1 microM), or the phosphodiesterase inhibitors, zaprinast (10-100 microM), dipyridamole (1-10 microM), or isobutyl methyl xanthine (IBMX, 0.1-0.5 mM). ACE amounts were measured by inhibitor binding assay and cellular cGMP levels by radioimmunoassay. RESULTS: ANP caused a dose dependent increase in ACE measured in intact endothelial cell culture. The stimulatory effect of ANP was blocked by Rp-8-Br-PET-cGMPS, a protein kinase G inhibitor. The cyclic GMP analog, 8-Br-cGMP and the cyclic GMP specific phosphodiesterase inhibitor, zaprinast, both increased ACE. Increase of ACE was also caused by nonspecific phosphodiesterase inhibitors, dipyridamole and IBMX. Intracellular cGMP levels were shown to increase by ANP, and phosphodiesterase inhibitors. CONCLUSIONS: These data suggest that cGMP is an intracellular mediator regulating ACE and that ANP induced increase of ACE is mediated via a cGMP dependent mechanism. PMID- 9876334 TI - Terbutaline-induced desensitization of human cardiac beta 2-adrenoceptor-mediated positive inotropic effects: attenuation by ketotifen. AB - BACKGROUND: In patients with chronic heart failure cardiac beta 1-adrenoceptors are desensitized whereas beta 2-adrenoceptors are only marginally affected. The mechanism underlying this differential regulation is not known. OBJECTIVES: To find out whether or not human cardiac beta 2-adrenoceptors might be 'resistant' to agonist-induced desensitization and whether or not the antiallergic drug ketotifen might attenuate possible desensitization. METHODS: We investigated, in a single blinded, randomised, placebo-controlled, cross-over study of ten healthy male volunteers (mean age, 25.3 +/- 0.7 years), the effects of two weeks treatment with the beta 2-adrenoceptor agonist terbutaline (3x5 mg/day p.o.) with and without simultaneous treatment with ketotifen (2x1 mg/day p.o. for three weeks) or placebo on beta-adrenoceptor-mediated cardiovascular effects. Cardiovascular effects were assessed as isoprenaline (3.5-35 ng/kg/min)- and terbutaline (25-150 ng/kg/min)-infusion-induced increases in heart rate and systolic blood pressure, decreases in diastolic blood pressure and shortening of the systolic time intervals (STIs), heart rate corrected duration of electromechanical systole (QS2c) and pre-ejection period (PEP; as a measure of inotropism). RESULTS: Ketotifen did not significantly affect basal haemodynamics in the volunteers. Isoprenaline- and terbutaline-infusion caused dose-dependent increases in systolic blood pressure and heart rate, decreases in diastolic blood pressure and shortening of QS2c and PEP, whereby isoprenaline effects were more pronounced. After two weeks of treatment with terbutaline p.o., isoprenaline- and terbutaline-infusion-induced increases in heart rate, shortening of QS2c and PEP were significantly reduced whereby terbutaline-infusion effects were markedly more attenuated than isoprenaline-infusion effects. Ketotifen significantly reduced terbutaline p.o. treatment-induced attenuation of all terbutaline infusion effects (largely beta 2-adrenoceptor-mediated) and the isoprenaline infusion-induced increase in heart rate (beta 1- and beta 2-adrenoceptor mediated), but did not (or only marginally) affect reduction in isoprenaline induced shortening of QS2c and PEP (largely beta 1-adrenoceptor-mediated). CONCLUSION: Human cardiac beta 2-adrenoceptors are not 'resistant' to agonist induced desensitization: Ketotifen might prevent such beta 2-adrenoceptor-agonist evoked desensitization. PMID- 9876335 TI - Platelet aggregation in acute coronary syndromes: use of a new aggregometer with laser light scattering to assess platelet aggregability. AB - OBJECTIVE: Platelet aggregation has been implicated in the pathogenesis of acute coronary syndromes. Small aggregates consisting of < or = 100 platelets cannot be quantified with a conventional aggregometer employing optical density. Using a recently developed aggregometer based on laser light scattering, we studied platelet aggregability in patients with acute coronary syndromes. METHODS: Peripheral blood samples were obtained from 39 patients with acute myocardial infarction or unstable angina who had received no prior antiplatelet or anticoagulant therapy, to be assayed immediately using a PA-100 platelet aggregometer. Blood samples from 14 healthy volunteers were used as controls. RESULTS: Spontaneous formation of platelet aggregates was observed only in patients with acute coronary syndromes. The size of these aggregates was small, consisting of < or = 100 platelets (primary aggregation). Agonist-induced aggregation consisted of two phases. In the first few minutes, the number of small aggregates increased markedly (primary aggregation), followed by an increase in larger aggregates (secondary aggregation). The EC50 of epinephrine for primary aggregation was nearly 50 times lower in acute coronary patients than in controls (P < 0.001), while the EC50 for secondary aggregation was only 2 times lower (P < 0.001). CONCLUSIONS: Aggregometry using light scattering suggests that platelet hyperaggregability and hypersensitivity in acute coronary syndromes may occur in primary but not secondary aggregation. PMID- 9876336 TI - Congenital adrenal hyperplasia due to steroid 21-hydroxylase deficiency. PMID- 9876337 TI - Octreotide and ectopic Cushing's syndrome--a commentary. PMID- 9876338 TI - The effect of six months treatment with a 100 mg daily dose of dehydroepiandrosterone (DHEA) on circulating sex steroids, body composition and muscle strength in age-advanced men and women. AB - OBJECTIVE: The biological role of the adrenal sex steroid precursors--DHEA and DHEA sulphate (DS) and their decline with ageing remains undefined. We observed previously that administration of a 50 daily dose of DHEA for 3 months to age advanced men and women resulted in an elevation (10%) of serum levels of insulin like growth factor-I (IGF-I) accompanied by improvement of self-reported physical and psychological well-being. These findings led us to assess the effect of a larger dose (100 mg) of DHEA for a longer duration (6 months) on circulating sex steroids, body composition (DEXA) and muscle strength (MedX). SUBJECTS AND DESIGN: Healthy non-obese age-advanced (50-65 yrs of age) men (n = 9) and women (n = 10) were randomized into a double-blind placebo-controlled cross-over trial. Sixteen subjects completed the one-year study of six months of placebo and six months of 100 mg oral DHEA daily. MEASUREMENTS: Fasting early morning blood samples were obtained. Serum DHEA, DS, sex steroids, IGF-I, IGFBP-1, IGFBP-3, growth hormone binding protein (GHBP) levels and lipid profiles as well as body composition (by DEXA) and muscle strength (by MedX testing) were measured at baseline and after each treatment. RESULTS: Basal serum levels of DHEA, DS, androsternedione (A), testosterone (T) and dihydrotestosterone (DHT) were at or below the lower range of young adult levels. In both sexes, a 100 mg daily dose of DHEA restored serum DHEA levels to those of young adults and serum DS to levels at or slightly above the young adult range. Serum cortisol levels were unaltered, consequently the DS/cortisol ratio was increased to pubertal (10:1) levels. In women, but not in men, serum A, T and DHT were increased to levels above gender-specific young adult ranges. Basal SHBG levels were in the normal range for men and elevated in women, of whom 7 of 8 were on oestrogen replacement therapy. While on DHEA, serum SHBG levels declined with a greater (P < 0.02) response in women (-40 +/- 8%; P = 0.002) than in men (-5 +/- 4%; P = 0.02). Relative to baseline, DHEA administration resulted in an elevation of serum IGF-I levels in men (16 +/- 6%, P = 0.04) and in women (31 +/- 12%, P = 0.02). Serum levels of IGFBP-1 and IGFBP-3 were unaltered but GHBP levels declined in women (28 +/- 6%; P = 0.02) not in men. In men, but not in women, fat body mass decreased 1.0 +/- 0.4 kg (6.1 +/- 2.6%, P = 0.02) and knee muscle strength 15.0 +/- 3.3% (P = 0.02) as well as lumbar back strength 13.9 +/- 5.4% (P = 0.01) increased. In women, but not in men, an increase in total body mass of 1.4 +/- 0.4 kg (2.1 +/- 0.7%; P = 0.02) was noted. Neither gender had changes in basal metabolic rate, bone mineral density, urinary pyridinoline cross-links, fasting insulin, glucose, cortisol levels or lipid profiles. No significant adverse effects were observed. CONCLUSIONS: A daily oral 100 mg dose of DHEA for 6 months resulted in elevation of circulating DHEA and DS concentrations and the DS/cortisol ratio. Biotransformation to potent androgens near and slightly above the range of their younger counterparts occurred in women with no detectable change in men. Given this hormonal milieu, an increase in serum IGF-I levels was observed in both genders but dimorphic responses were evident in fat body mass and muscle strength in favour of men. These differences in response to DHEA administration may reflect a gender specific response to DHEA and/or the presence of confounding factor(s) in women such as oestrogen replacement therapy. PMID- 9876339 TI - Growth hormone and prolactin responses to corticotrophin-releasing-hormone in patients with Cushing's disease: a paracrine action of the adenomatous corticotrophic cells? AB - OBJECTIVE: In patients with Cushing's disease multihormonal responses to ovine corticotrophin releasing hormone (oCRH) have been detected in blood from inferior petrosal sinuses. This finding has been explained by co-secretion of other hormones, in addition to ACTH, by the pituitary adenoma itself or by paracrine effects exerted by the adenoma on normal periadenomatous pituitary cells. To assess these hypotheses we compared the presence of a CRH induced GH and/or PRL response during inferior petrosal sinus sampling to the immunohistochemical detection of PRL and GH in adenomatous tissue removed from patients with Cushing's disease. PATIENTS AND MEASUREMENTS: Twenty-two patients with Cushing's disease and two patients with ectopic ACTH syndrome due to a bronchial carcinoid were studied; each patient had undergone preoperative inferior petrosal sinus sampling for diagnostic purposes with determination of GH and PRL in addition to ACTH, before and after administration of oCRH. Immunohistochemical studies for ACTH, GH and PRL detection were carried out on adenomatous tissue removed at surgery in the patients with pituitary dependent Cushing's disease and on the carcinoid tumours from the two patients with ectopic ACTH syndrome. RESULTS: All pituitary adenomas immunostained for ACTH, and four adenomas immunostained for GH or PRL in addition to ACTH. A PRL increase in the inferior petrosal sinus after oCRH administration was found in 11 of 22 patients, but none of their tumours immunostained for PRL. Immunostaining for PRL was found in the pituitary tumours from two patients but in neither patient was there a PRL response after oCRH. A GH response was found in 13 of 20 patients in whom it was sought; no patient showed immunostaining in their tumour. GH immunostaining was found in two tumours but in neither patient was there a GH response after oCRH. The oCRH-induced increase of GH and PRL was always recorded in the dominant inferior petrosal sinus. The ACTH response to oCRH was significantly higher in patients who showed oCRH induced increases in GH and/or PRL than in patients who did not, both in terms of area under the response-curve (22,032 +/- 9876 vs. 4371 +/- 2870 ng/l/10 min; P < 0.05) or mean percentage increase above baseline (754 +/- 229% vs. 147.2 +/- 67%, P < 0.02). A significant correlation was observed between ACTH and GH responses to oCRH. The two patients with ectopic Cushing's syndrome did not show ACTH, GH or PRL increases after oCRH administration and did not show immunostaining for GH or PRL in their tumours. CONCLUSIONS: The present data do not support the hypothesis of co-secretion of hormones by the pituitary adenoma as the cause of the GH and PRL responses to ovine corticotrophin releasing hormone observed in patients with Cushing's disease; it is suggested that a different mechanism, possibly involving an interaction between the ACTH secreting adenoma and the normal periadenomatous GH and PRL secreting cells, may be responsible. PMID- 9876340 TI - Increased incidence of neoplasia in patients with pituitary adenomas. The Pituitary Study Group. AB - OBJECTIVE: The goal of our study was to determine the rate of neoplasms in patients with other pituitary adenomas (non-functioning and prolactinomas) in comparison with acromegaly which is known to favour the development of neoplasia. DESIGN AND PATIENTS: We reviewed clinical records for 220 patients with acromegaly, 151 patients with non-functioning pituitary adenoma (NF) and 98 patients with prolactinomas. Incidence rates of cancer for patients with pituitary tumours were calculated per person-years of follow-up study. These rates were then compared with sex and age adjusted incidence rates reported by National Tumour Registry. An internal control group of 163 subjects with a non neoplastic condition, i.e. Graves' disease followed chronically in the same clinic was also studied. The ratios observed to expected were expressed as standardized incidence rates (SIR). The only significant difference between the acromegalic and other pituitary tumours patients was in hypopituitarism, present in 18.2% (acromegaly) 47% (NF) and 18.6% (prolactinomas). RESULTS: Twenty-three malignant tumours were registered in 19 acromegalics (1 Hodgkin disease, 1 myelogenous leukaemia, 1 lymphocytic leukaemia, 3 papillary thyroid carcinomas, 1 ovarian carcinoma, 2 colorectal carcinoma, 1 renal cell carcinoma, 4 cervical carcinoma, 2 skin cancers, 2 pancreatic carcinoma, 4 breast carcinoma, 1 bladder carcinoma). Three acromegalics harboured two malignancies. Patients with acromegaly had a 3.39-fold increased rate of malignant tumours compared with the general population and a 3.21-fold increased rate compared with our internal control group. Eleven malignant tumours were found in patients with NF-pituitary adenomas and 2 in prolactinoma patients (1 lymphoma, 1 multiple myeloma, 1 colonic cancer, 1 renal cell cancer, 1 stomach cancer, 2 lung cancers, 1 cervix carcinoma, 1 breast cancer, 1 testicular carcinoma and 3 melanoma). Patients with NF pituitary adenomas had a 3.91-fold increased rate of malignant tumours compared with the general population and 4.07-fold increase compared with the internal control group. Patients harbouring prolactinomas did not have an increased incidence rate of malignancy compared with the general population or our internal controls. Female patients with acromegaly and male patients with NF pituitary adenoma had higher incidences of neoplasia. CONCLUSION: We have demonstrated that the overall incidence of malignant tumours in patients with non functioning pituitary adenomas and acromegaly is significantly higher than expected for general population and for our internal control group. PMID- 9876341 TI - The impact of obesity on cardiovascular risk factors in Turner's syndrome. AB - OBJECTIVE: To evaluate risk factors for coronary artery disease in women with Turner's syndrome which may contribute to the increased incidence of premature cardiovascular death noted in this disorder. DESIGN: Comparison of clinical and biochemical parameters in women with Turner's syndrome with those in women with normal karyotype. PATIENTS: Ninety-one women with Turner's syndrome attending a dedicated adult Turner's syndrome clinic and 22 control subjects were studied. MEASUREMENTS: Recumbent blood pressure, body mass index (BMI), fasting total cholesterol, triglycerides and high density lipoproteins (HDL) were measured in both study groups. RESULTS: Women with Turner's syndrome were more obese compared with women with a normal karyotype. They were more likely to be hypertensive and the obese patients had higher serum triglyceride concentrations. Hypertension was independent of obesity and may be under-recognized because of failure to compare with age-matched normal ranges. Lipoprotein changes were accounted for by the obesity. CONCLUSIONS: Women with Turner's syndrome may be at increased risk of developing coronary artery disease as a result of the higher frequency of hypertension and obesity. Routine screening of this population for risk factors for ischaemic heart disease is recommended. PMID- 9876342 TI - Is there a methimazole dose effect on remission rate in Graves' disease? Results from a long-term prospective study. The European Multicentre Trial Group of the Treatment of Hyperthyroidism with Antithyroid Drugs. AB - OBJECTIVE: The optimal antithyroid drug regimen for Graves' disease remains a matter of controversy. The European Multicentre Trial Group has investigated the effects of methimazole drug dose on the long-term outcome of Graves' disease. DESIGN: Extended follow-up of patients from a prospective multicentre trial, designed to study methimazole dose effects on the outcome of Graves' disease. We have reported previously that the relapse rates did not differ after a medication free observation period of 12 months; the relapse rates were 37% and 38%, respectively. In this paper, we describe the outcome in these patients after a mean observation period of 4.3 +/- 1.3 years and have looked for potential predictors of this outcome. PATIENTS: Three hundred and thirteen patients with Graves' disease were randomized to treatment with a constant dose of 10 or 40 mg of methimazole for 1 year, with levothyroxine supplementation as required. MEASUREMENTS: At the time of inclusion into the trial: thyroid size, T4, T3, TSH binding inhibiting immunoglobulins, urinary iodide excretion, thyroid uptake, Crook's therapeutic index of hyperthyroidism (a measure of clinical disease severity). At the time of follow-up examination: TSH, T4, T3, thyroid size, thyroid ultrasound, THS-binding inhibiting immunoglobulins. RESULTS: The overall relapse rate was 58%. There was no difference in relapse rates between patients treated with either 10 or 40 mg of methimazole (58.3 vs. 57.8%). Five patients had become spontaneously hypothyroid, without obvious relationship to antithyroid drug dose. Patients who relapsed and patients who remained in remission did not differ with respect to: age, goitre size, ophthalmopathy, median iodine excretion, serum T4 or serum T3, Crook's therapeutic index and thyroid uptake at the time of study entry. Thus, none of these variables was potentially suitable for predicting outcome. This finding was confirmed by Cox's proportional hazard regression. Thyroid volume, measured by ultrasound, did not differ between patients in remission and patients with relapse. There was no difference in the course of endocrine eye signs, in the requirement for steroid and radiotherapy for eye signs, or in thyroid echostructure between patients in the 10 and in the 40 mg group, nor was serum TSH different in patients who had remained in remission (0.8 +/- 0.6 mU/l in the 10 mg group, 1.0 +/- 0.8 mU/l in the 40 mg group). CONCLUSIONS: The dose of methimazole in Graves' disease therapy can safely be kept to the minimal required dose. This will provide the same chance of remission as higher doses, and provide the best balance of risk and benefit. PMID- 9876343 TI - Growth hormone induced increase in serum IGFBP-3 level is reversed by anabolic steroids in substance abusing power athletes. AB - OBJECTIVE: Body builders may use growth hormone (GH) and anabolic steroids to increase fitness and muscle mass. GH increases serum concentrations of IGF-I and IGFBP-3. The combined effects of GH and anabolic androgenic steroids on IGFBP-3 and IGF-I levels are not known under authentic substance-abusing conditions. The aim of this study was to investigate this in substance-abusing power athletes. DESIGN AND PATIENTS: Nine healthy, non-obese and non-competing male power athletes, all aggressive substance abusers, used GH and/or anabolic steroids independently of this study. Blood samples were taken both during and between the drug intake. Sixteen substance non-abusing wrestlers served as controls. MEASUREMENTS: Serum IGF-I concentration was measured by radioimmunoassay and the IGFBP-3 concentration was measured by two immunofluorometric assays, one detecting proteolytic fragments of IGFBP-3. The capacity of serum to proteolytically cleave IGFBP-3 was studied by the proteolysis assay. RESULTS: While growth hormone increased the IGFBP-3 and IGF-I concentrations, anabolic steroids decreased the same. Concomitant use of growth hormone and anabolic steroids decreased the IGFBP-3 concentration in five out of six abuse periods in spite of the fact that the IGF-I concentration remained elevated in four of them. However, in two men who were on low calorie diet both the IGF-I and IGFBP-3 concentrations decreased during combined GH/anabolic steroid abuse. No proteolytic fragmentation of IGFBP-3 was observed. CONCLUSION: Massive abuse of anabolic steroids decreases both the basal and GH-stimulated IGFBP-3 concentrations, whereas its effects on serum IGF-I concentration are variable and affected by low calorie diet. This study demonstrates that detection of GH doping by measuring the IGF-I and/or IGFBP-3 levels has notable confounding factors. PMID- 9876344 TI - The IGF-I/IGFBP system in congenital partial lipodystrophy. AB - BACKGROUND AND OBJECTIVES: Insulin and IGF-I interact at many levels. Little is known about the insulin-like growth factor-I/insulin-like growth factor binding proteins (IGF-I/IGFBP) system in congenital partial lipodystrophy, a syndrome characterized by insulin resistance, hyperinsulinaemia and absence of truncal and limb fat. Some cases have acromegaloid features with thick skin and large hands and feet in association with normal levels of circulating growth hormone. METHODS: In four females known with congenital partial lipodystrophy, hyperinsulinaemia with acromegaloid features, the number and affinity of the IGF I receptors on peripheral blood mononuclear cells (PBMCs), and the concentration of circulating insulin, total and free IGF-I, IGFBP-1 and IGFBP-3 levels were measured in the fasting and the fed state. Cultures of PBMCs of the patients with lipodystrophy were also used to study the effect of IGF-I stimulation on thymidine uptake in vitro. MEASUREMENTS: In the subjects with lipodystrophy the affinity and the number of the IGF-I receptors on peripheral mononuclear cells (PBMCs) and erythrocytes did not differ significantly from controls in the fasting state. Insulin levels were significantly higher in subjects with lipodystrophy both in the fasting as well in the fed state. Total IGF-I, free IGF I and IGFBP-3 levels did not differ but serum IGFBP-1 levels were lower in lipodystrophy subjects than in healthy controls. The free IGF-I/IGFBP-1 ratio was increased in lipodystrophy subjects both in the fasting and the fed states. The effects of IGF-I stimulation on thymidine uptake by PBMCs of lipodystrophy subjects in the absence of IGFBP-1 were not different from healthy controls cultures in vitro. When a combination of IGFBP-1 (in a concentration comparable to the fasting serum IGFBP-1 levels in lipodystrophy patients found in our study) and IGF-I was added to PBMC cultures from lipodystrophy patients no decrease in thymidine uptake by PBMCs was found. CONCLUSIONS: In the four subjects with lipodystrophy hyperinsulinaemia, lowered free IGF-I and IGFBP-1 levels, but increased free IGF-I/IGBP-1 ratios were observed. Low IGFBP-1 concentrations in culture media did not reduce the stimulating IGF-I effect on thymidine uptake by PBMCs from lipodystrophy patients. Our data suggest that the observed increased IGF-I/IGFBP-1 ratio in lipodystrophy patients contributes to an unopposed biological effect of IGF-I on IGF-I receptors, thereby inducing the development of acromegaloid features, acanthosis nigricans and polycystic ovaries in some patients with congenital partial lipodystrophy. PMID- 9876345 TI - Overexpression of vasopressin (V3) and corticotrophin-releasing hormone receptor genes in corticotroph tumours. AB - OBJECTIVE: The molecular mechanisms underlying ACTH-secreting tumour formation remain unknown. Transmembrane signalling pathways play an important role in several endocrine disorders including pituitary tumours. To investigate the role of the pituitary vasopressin (V3) receptor (R) in ACTH-secreting tumours we have qualitatively and quantitatively analysed its mRNA. DESIGN: RT-PCR, denaturing gradient gel electrophoresis and S1 nuclease protection experiments were used to analyse V3 mRNA structure in ACTH-secreting tumours. We also developed a competitive RT-PCR system to compare the levels of expression of POMC, V3 and CRH R genes. This system used as competitor a single mutant template (termed multi mutant) containing primers for the three genes flanking an unrelated core sequence allowing multiple quantifications from the same cDNA preparations. We analysed 12 normal pituitaries, 15 corticotroph pituitary adenomas and 6 ACTH secreting bronchial carcinoids. RESULTS: The V3 mRNA structure and sequence were found to be identical in normal and tumoural pituitary indicating that the tumoural Vs mRNA codes for a normal receptor. POMC RT-PCR signals in the pituitary tumour group were approximately 7-fold higher than in the normal pituitary group. Similarly, V3 and CRH-R signal were increased in pituitary tumors (mean +/- SEM: 5.87 x 10(-6) +/- 1.73 x 10(-6), and 2.33 x 10(-4) +/- 1.4 x 10(-4), respectively), when compared to normal pituitaries (1.19 x 10(-7) +/- 2.39 x 10(-8), and 1.7 x 10(-6) +/- 4.65 x 10(-7), respectively) suggesting that these two genes are expressed at very high levels in corticotroph tumours. When expressed relative to the corresponding POMC signals, increases in V3 and CRH-R signals reached 49-fold and 137-fold, respectively, in pituitary tumours. In ACTH secreting bronchial carcinoids V3 gene expression level was also higher than in normal pituitary, whereas CRH-R signals were detected in only 4 of the 6 tumours with wide variations. CONCLUSION: Our results show that both vasopressin and CRH receptor genes are overexpressed in ACTH-secreting pituitary tumours. They suggest that overexpression of G protein-coupled receptors may be an additional mechanism through which membrane receptors may play a role in human tumours. PMID- 9876346 TI - Responsiveness of IGF-I and IGFBP-3 to therapeutic intervention in children and adolescents with Crohn's disease. AB - OBJECTIVE: Abnormal linear growth is common in childhood and adolescent Crohn's disease. We have studied the concentrations of the inflammatory marker CRP and of serum IGF-I and IGFBP-3 in patients with active Crohn's disease and have assessed the changes in these parameters during therapeutic intervention with enteral nutrition or intestinal resection. DESIGN: Children and adolescents attending the inflammatory bowel disease clinic at our hospital underwent treatment either with enteral nutrition (Study A) or intestinal resection (Study B). These are two separate studies and the results cannot be compared. Serum concentrations of CRP, IGF-I and IGFBP-3 were determined at 0, 2, 8 and 16 weeks after start of enteral nutrition and in addition to height velocity, at 0 and 6 months after intestinal resection. SUBJECTS: Study A: 14 patients, 9 male, 5 female, median age 12.5 years (range 7.0-17.2), puberty stage 1 (n = 13), stage 3 (n = 1). All had active Crohn's disease. Study B: 9 patients, 7 male, 2 female, median age 13.5 years (range 7.8-16.5), puberty stage 1 (n = 5), stages 2-4 (n = 4). All had Crohn's disease resistant to medical therapy. METHODS: Crohn's disease was confirmed radiologically, endoscopically and histologically. Disease activity was scored using the Lloyd Still index (LSI). Study A: nutritional support was with a polymeric, casein-based formula feed AL 110. Study B: surgical procedures were small bowel resection (n = 2), right hemicolectomy (n = 5), subtotal colectomy (n = 2). MEASUREMENTS: Study A: weight SDS, CRP, IGF-1 and IGFBP-3 were measured at 0, 2, 8, 16 weeks after start of enteral feeding. Study B: height velocity, CRP, IGF-I and IGFBP-3 were measured 0, 6 months after intestinal resection. STATISTICAL ANALYSIS: Medians and ranges were used. Significance of changes was calculated using the Wilcoxon rank test for the analysis of paired data. RESULTS: Study A: median LSI before treatment was 39 and increased after 8 weeks of enteral nutrition to 60 (P < 0.05). Weight SDS increased at 8 and 16 weeks (P < 0.05) compared to pretreatment. CRP was elevated at 0 weeks, falling during treatment. Median (range) values (normal < 5 mg/l) at 0 at 2, 8, 16 weeks were 53 mg/l (15-150), 8 mg/l (5-25), 7 mg/l (5-83) and 14 mg/l (5-39), all P < 0.001 compared with pretreatment. Median IGF-I-values increased during treatment. Median (range) values at 0, 2, 8, 16 weeks (all P < 0.005) compared to pretreatment, median (range) values at 0, 2, 8, 16 weeks were 78 micrograms/l (50 204), 131 micrograms/l (73-251), 119 micrograms/l (77-291) and 133 micrograms/l (67-497), all P < 0.005 compared to pre-treatment. IGFBP-3 levels increased during treatment. Median (range) values at 0, 2, 8, 16 weeks were 2.4 mg/l (1.4 3.1), 2.9 mg/l (1.8-4.6), 3.0 mg/l, 3.2 mg/l (1.8-4.5), all P < 0.01 compared to pretreatment. Study B: height velocity increased during 6 months after surgery. Median (range) values; 3.3 cm/year (0-8.3) before surgery, 8.4 cm/year (2-12.6) 6 months post-surgery, P < 0.01. Median (range) CRP values fell from 45 mg/l (5 150) to 8 mg/l (5-31) and IGF-I-values increased from 163 micrograms/l (64-286) to 226 micrograms/l (71-391). These changes were not statistically significant. IGFBP-3 values did not change. CONCLUSION: The IGF system, as shown by serum IGF I and IGFBP-3, is responsive to therapeutic intervention in active Crohn's disease. It is likely that a combination of decreased inflammatory activity and improved nutrition contributes to these changes. PMID- 9876347 TI - Glucocorticoid therapy suppresses abnormal secretion of big IGF-II by non-islet cell tumours inducing hypoglycaemia (NICTH) AB - OBJECTIVE: To assess the relative efficacy of hGH and glucocorticoids in the treatment of non-islet cell tumour hypoglycaemia (NICTH) by examination of their influence on the composition of the various molecular species involving tumour and mature forms of IGF-II in association with IGFBP-3. DESIGN: Two groups each of 4 patients, all diagnosed as cases of NICTH, were treated with either hGH or glucocorticoids. Through the use of acidic size exclusion chromatography serum levels of tumour (big) and mature IGF-II were evaluated. Neutral size exclusion chromatography was used in the separation of molecular species before assay for immunoreactive IGF-II and IGFBP-3 content. RESULTS: High-dose hGH treatment produced increases in serum levels of big and mature IGF-II and IGFBP-3 but without generation of high molecular weight complexes. Glucocorticoid treatment suppressed big IGF-II permitting re-establishment of normal IGF/IGFBP association patterns. CONCLUSION: Glucocorticoid therapy has been demonstrated to consistently reverse the biochemical abnormalities caused by tumour-derived big IGF-II compared with the potentially adverse stimulatory effects of hGH treatment in causing increases in serum levels of big IGF-II. PMID- 9876348 TI - Is opioidergic activity responsible for the circadian variation observed in the gonadotrophin responsiveness of early follicular phase women? AB - OBJECTIVES: In women, the gonadotrophin response to gonadotrophin-releasing hormone (GnRH) displays a circadian rhythm during the early follicular phase (EFP), with GnRH-stimulated luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release found to be markedly decreased at night. Since the opioidergic inhibition of gonadotrophin secretion is selectively enhanced at night, we reasoned that the circadian changes in the gonadotrophin responsiveness to GnRH might be related to a nocturnal increase of opioidergic activity. STUDY DESIGN: Eleven women with normal menstrual cycles were studied in the EFP on four different occasions in random order. Studies were conducted either during the day (0900-1300 h) or at night (2100-0100 h). During these times, GnRH (25 micrograms i.v.) was administered in conjunction with either saline (as control) or naloxone (4 mg i.v.). MEASUREMENTS: Frequent blood samples were obtained before and after GnRH stimulation for determination of basal sex steroid and gonadotrophin concentrations by immunoradiometric assays. RESULTS: While oestradiol levels were comparable (P > 0.3) at all times, progesterone concentrations were significantly (P < 0.01) higher during day than during night hours, with no difference between control and naloxone conditions. Gonadotropin responses to GnRH stimulation were not significantly different between day and night times, nor did they vary between control and naloxone conditions. CONCLUSIONS: Opioidergic blockade imposed by naloxone did not noticeably change GnRH-stimulated gonadotrophin release at any of the study times. We therefore infer that mechanisms other than a nocturnal increase of opioidergic inhibition may account for eventual circadian changes in the gonadotrophin responsiveness of early follicular phase women. PMID- 9876349 TI - Leptin is involved in gender-related differences in insulin sensitivity. AB - BACKGROUND: Although the effects of insulin on leptin levels are relatively well characterized, the possible actions of leptin on insulin sensitivity are not so well studied. This study was undertaken to examine whether gender-related differences in insulin sensitivity could be explained partially by leptin levels. SUBJECTS: The study involved 22 women (13 obese) and 20 (11 obese) fat mass- and age-matched men. All participants were healthy. MEASUREMENTS: Several anthropometric measures of body fatness were quantified and the percentage of body fat was determined through bioelectric impedance. Oral glucose tolerance test and a frequently sampled intravenous glucose tolerance test was performed in all subjects. Serum leptin was measured by radioimmunoassay. RESULTS: Nine lean women (BMI 20.4 +/- 2 kg m2 mean +/- SD) showed increased leptin levels (7.8 +/- 2.7 vs. 4.3 +/- 1.3 micrograms/l, P = 0.003), increased insulin sensitivity (5.2 +/- 1.3 vs. 2.9 +/- 0.9 min-1/mU/l, P = 0.001) and similar fat mass (11.1 +/- 3.7 vs. 13.2 +/- 7.8 kg, P = NS) in comparison with 9 age-matched lean men (33.6 +/- 6 vs. 34.5 +/- 6.3 years, P = NS). Thirteen obese women (BMI 32.5 +/- 2.7) kg m2 also showed increased leptin levels (29.6 +/- 8.4 vs. 11.7 +/- 4.8 micrograms/l, P < 0.0001), increased insulin sensitivity (1.7 +/- 0.7 vs. 0.95 +/- 0.9 min-1 mU/l, P = 0.04) and similar fat mass (34.4 +/- 8.0 vs. 30.9 +/- 9.6 kg, P = NS) in comparison with 11 age-matched obese men (34.5 +/- 7.8 vs. 38.7 +/- 8.2 years, P = NS). A strong linear association between leptin levels and insulin sensitivity (Si) was found (r = -0.67, P = 0.001, in men; r = -0.82, P < 0.0001, in women). After controlling for percentage of body fat, this association remained significant only in men (r = -0.56, P = 0.01, in men; r = -0.30, P = NS in women). In stepwise regression analysis models, both gender (P = 0.00001) and leptin (P = 0.00001) contributed to 67% of the variance in Si independently of body fat. CONCLUSIONS: Leptin levels and gender contribute to the variance of insulin sensitivity, independently of body fat. These results suggest that leptin could affect insulin sensitivity. PMID- 9876350 TI - Serum prolactin concentration in patients taking neuroleptic drugs. AB - OBJECTIVE: Non-tumour causes of hyperprolactinaemia, including prolactin elevating drugs, must be excluded. There is a general view that such drugs are unlikely to raise serum PRL above 3000 mU/I, but the literature is confusing. We report 8 patients receiving treatment with neuroleptic drugs, whose serum PRL concentrations were grossly elevated. METHODS: Prolactin was measured using a 2 site immunofluorometric assay (Abbott Laboratories; reference range < 500 mU/l). Seven of the eight women (age range 24-49 years) were symptomatic (galactorrhoea, oligo- or amenorrhoea). RESULTS: Prolactin concentrations ranged from 3600 mU/l to 7300 mU/l. All patients had a normal pituitary CT scan. Five patients were treated with bromocriptine without detriment to their mental state. CONCLUSION: Prolactin can rise to concentrations associated with prolactinomas in patients on neuroleptic drugs. As it is rarely possible to stop the drugs to see if the PRL concentration will decline to normal, neuroradiology is required in these patients to exclude a vision-threatening macroprolactinoma before deciding on medical treatment. PMID- 9876351 TI - Regulation and tissue distribution of the human sodium iodide symporter gene. AB - OBJECTIVE: Iodide uptake by the thyroid gland is mediated by the sodium iodide symporter (NIS). In the present report, we have analysed the factors that modulate human NIS mRNA expression and iodide uptake in primary thyroid follicular cell (TFC) cultures. In addition, NIS mRNA tissue distribution was investigated. METHODS: Primary thyroid follicular cell cultures were treated with human recombinant TSH with or without cytokines for 72 h. Subsequently, NIS gene expression and iodide uptake were analysed using reverse transcription-polymerase chain reaction (RT-PCR) and 125I uptake, respectively. Human tissue samples were investigated for NIS gene expression using both RT-PCR and Northern blotting. RESULTS: Human TSH increased both NIS gene expression and iodide uptake in TFC cultures in a dose-dependent manner. Using concentrations of 0.1 U/l of hTSH, a minor increase in NIS gene expression was detected without a detectable increase in iodide uptake. IL-1 alpha, TNF alpha and IFN gamma at concentrations of 10(5) U/l all inhibited TSH-induced NIS gene expression and iodide uptake. In these experiments, there was a good correlation between NIS mRNA expression and iodide uptake. Using RT-PCR higher levels of NIS mRNA were detected in Graves' disease (GD) compared to multi-nodular goitre tissue samples. Stomach and salivary gland tissue also expressed NIS mRNA, whereas low levels were found in the mammary gland and extraocular muscle tissue. No expression was detected in the ovary, oesophagus, colon, extraocular fat or skin. In contrast, Northern blot analysis failed to detect NIS in stomach, salivary gland, intestinal fat or non-toxic multi-nodular goitre tissue samples, although this was present in GD thyroid tissue. CONCLUSION: TSH upregulates sodium iodide symporter gene expression and iodide uptake in primary thyroid follicular cell cultures, and this induction is modulated by cytokines. Variable levels of sodium iodide symporter mRNA are present in different tissue samples, with high expression evident in Graves' disease thyroid tissue. PMID- 9876352 TI - GNAS1 mutational analysis in pseudohypoparathyroidism. AB - OBJECTIVE: Mutations of the GNAS1 gene, which is located on chromosome 20q13.11 and encodes the alpha-subunit of the stimulatory GTP-binding protein, have been identified in patients with pseudohypoparathyroidism type Ia (PHPIa) and pseudopseudohypoparathyroidism (PPHP). We have undertaken studies to determine the prevalence of GNAS1 mutations and to explore methods for their more rapid detection. METHODS: Thirteen unrelated families (8 with PHPIa and PPHP patients, and 5 with PPHP patients only) were investigated for GNAS1 mutations in the 1050 base-pair (bp) region spanning exons 2-13 by single-stranded conformational polymorphism (SSCP) and DNA sequence analysis. RESULTS: GNAS1 mutations were detected in 4 of the 8 families with PHPIa patients. These consisted of: two novel de novo missense mutations (Pro115Ser and Glu259Val) in two families and an identical 4 bp deletion of codons 189 and 190 resulting in a frame-shift in two unrelated families. These results expand the spectrum of GNAS1 mutations associated with this disorder and confirm the presence of a mutational hot-spot involving codons 189 and 190. SSCP analysis was found to be a specific and sensitive method that detected all 4 mutations. GNAS1 mutations were not detected in any of the PPHP only families. CONCLUSIONS: The pseudohypoparathyroid disorders appear to represent a heterogeneous group with GNAS1 mutations forming the molecular aetiology in approximately 50% of pseudohypoparathyroidism type Ia families. Such mutations can be reliably identified by single-stranded conformational polymorphism and this will help to supplement the clinical evaluation of some patients and their families, particularly as the disease may not be fully penetrant. PMID- 9876353 TI - A study of patients with Nelson's syndrome. AB - OBJECTIVE: The prevalence of Nelson's syndrome has varied greatly, at least in part because of the variability of the diagnostic criteria employed by different authors. We define Nelson's syndrome as the presence of an enlarging pituitary tumour associated with elevated fasting plasma ACTH levels and hyperpigmentation in patients with Cushing's disease after bilateral adrenalectomy. We have compared patients with Cushing's disease who developed Nelson's syndrome after bilateral adrenalectomy with those who did not. Our objective was to find differences between the two groups which might predict the development of Nelson's syndrome. PATIENTS AND METHODS: We have reviewed the records of 30 patients with Cushing's disease after adrenalectomy, and divided them into two groups; I: 14 who developed Nelson's syndrome and II, 16 who did not. The two groups of patients were compared in their clinical, laboratory and imaging data as well as in the therapeutic procedures that preceded the adrenalectomy. RESULTS: The comparison between the two groups of patients demonstrated a highly significant difference in relation to the development of cutaneous hyperpigmentation (100% in group I and 19% in group II) and neuro ophthalmological symptoms (21% in group I and 0% in group II) after adrenalectomy. There were no significant differences in laboratory data before adrenalectomy. After adrenalectomy, plasma ACTH levels increased significantly in the patients of both groups, but to much higher levels in those who developed Nelson's syndrome. Plasma ACTH concentrations above 154 pmol/l occurred only in the subjects with Nelson's syndrome. Before adrenalectomy, a pituitary tumour was more frequent in the patients who developed Nelson's syndrome (55% vs. 33% at transsphenoidal pituitary exploration). Pituitary surgery and irradiation were undertaken before adrenalectomy in approximately equal numbers of patients in each group. DISCUSSION: The prevalence of Nelson's syndrome was 47% in our series of 30 patients with Cushing's disease after bilateral adrenalectomy. No clinical or laboratory data before adrenalectomy predicted the development of the syndrome. The value of prophylactic pituitary irradiation could not be evaluated from our clinical material. However, after adrenalectomy, the presence of hyperpigmentation and ACTH levels above 154 pmol/l had positive predictive value for the development of Nelson's syndrome. In this situation magnetic resonance imaging (MRI) of the pituitary is mandatory and, if no tumour is detected, MRI should be repeated at intervals. PMID- 9876354 TI - Occult ectopic ACTH secretion due to recurrent lung carcinoid: long-term control of hypercortisolism by continuous subcutaneous infusion of octreotide. AB - We report on a woman with clinical Cushing's syndrome confirmed by biochemical data. The Cushing's syndrome was shown to be ACTH dependent and inferior petrosal sinus sampling pointed to an ectopic source. After resection of a lung carcinoid a well documented remission of Cushing's syndrome was obtained. At recurrence of Cushing's syndrome 18 months later the ACTH source could not be located despite extensive diagnostic procedures. Clinical and biochemical control of hypercortisolism was achieved by continuous subcutaneous infusion of octreotide. During a brief interruption of treatment recurrence of clinical and biochemical signs and symptoms of Cushing's syndrome were demonstrated. We conclude that in this case of occult ectopic ACTH secretion, presumably due to recurrent lung carcinoid, continuous subcutaneous infusion therapy with octreotide resulted in clinical and metabolic control of Cushing's syndrome for 8 years. In addition excellent tumour growth control was achieved as repeated searches for tumour recurrence or metastasis remained negative. PMID- 9876355 TI - Three cases of a hitherto unrecognized pituitary tumour: it is caused by maternal exposure to unidentified factor(s) during pregnancy? PMID- 9876356 TI - Our healthier nation: alpha for presentation, beta minus for deliverability. PMID- 9876357 TI - Psychosocial risk factors and mortality: a prospective study with special focus on social support, social participation, and locus of control in Norway. AB - STUDY OBJECTIVE: The objective is to investigate the effect on mortality of psychosocial variables, with special focus on social support, social participation, and locus of control. DESIGN: The study is designed as a prospective study with a 17 year follow up period, using univariate and multivariate proportional hazards regression analysis to estimate the predictive power of psychosocial variables, when controlling for sociodemographic and biological factors. SETTING: The study is based on a population sample randomly drawn from different neighbourhoods of Oslo in 1975/76, for the purpose of surveying health, in particular mental health, in relation to various social and psychosocial variables. The initial data were gathered by structured interviewing, whereas the data about mortality and cause of death, was gathered from the Central Bureau of Statistics. PARTICIPANTS: The initial sample included 1010 persons above the age of 18 years, with no upper age limit. The follow up with respect to mortality covered the whole sample, with the exception of a very few who had left the country. MAIN RESULTS: When controlling for socio demographic and biological factors, low social participation, and to a lesser extent, few close relationships and external locus of control, were associated with increased mortality. CONCLUSION: The effect of social participation and locus of control may indicate that life style, and individual psychological resources, are at least as important for survival as support from others in stressful life situations. PMID- 9876358 TI - Excess mortality in England and Wales, and in Greater London, during the 1995 heatwave. AB - STUDY OBJECTIVE: To assess the impact on mortality of the heatwave in England and Wales during July and August 1995 and to describe any difference in mortality impact between the Greater London urban population and the national population. DESIGN: Analysis of variation in daily mortality in England and Wales and in Greater London during a five day heatwave in July and August 1995, by age, sex, and cause. SETTING: England and Wales, and Greater London. MAIN RESULTS: An estimated 619 extra deaths (8.9% increase, approximate 95% confidence interval 6.4, 11.3%) were observed during this heatwave in England and Wales, relative to the expected number of deaths based on the 31-day moving average for that period. Excess deaths were apparent in all age groups, most noticeably in women and for deaths from respiratory and cerebrovascular disease. Using published daily mortality risk coefficients for air pollutants in London, it was estimated that up to 62% of the excess mortality in England and Wales during the heatwave may be attributable to concurrent increases in air pollution. In Greater London itself, where daytime temperatures were higher (and with lesser falls at night), mortality increased by 16.1% during the heatwave. Using the same risk coefficients to estimate the excess mortality apparently attributable to air pollution, more than 60% of the total excess in London was apparently attributable to the effects of heat. CONCLUSION: Analysis of this episode shows that exceptionally high temperatures in England and Wales, though rare, do cause increases in daily mortality. PMID- 9876359 TI - Urban-rural variations in health in The Netherlands: does selective migration play a part? AB - STUDY OBJECTIVE: Urban-rural health differences are observed in many countries, even when socioeconomic and demographic characteristics are controlled for. People living in urban areas are often found to be less healthy. One of the possible causes for these differences is selective migration with respect to health or health risk factors. This hypothesis is hardly ever empirically tested. This paper tries to assess the existence of selective urban-rural migration. DESIGN: Health indicators and health risk factors were measured in a 1991 population sample. Moves were registered between 1991 and 1995. Using logistic regression analyses, comparisons were made between, firstly, urban to rural movers and rural to urban movers and secondly, between movers and stayers. SETTING: Region surrounding the city of Eindhoven in south eastern part of the Netherlands. SUBJECTS: Data were used of 15,895 respondents aged 20-74 in 1991. By 1995 613 subjects had moved from urban to rural and 191 subjects from rural to urban. MAIN RESULTS: Bivariate nor multivariate analyses show hardly and differences between movers into urban and movers into rural areas. Bivariate analyses on movers and stayers show that movers are healthier than stayers. However, when socioeconomic and demographic variables are controlled for, movers appear to be less healthy, with the exception of the younger age groups. CONCLUSIONS: Areas that attract many migrants from and lose few migrants to other degrees of urbanicity will in the long run obtain healthier populations, because of demographic and socioeconomic characteristics. However, if these characteristics are accounted for, the opposite is true, with the exception of younger age groups. In extreme cases this may cause spurious findings in cross sectional research into the relation between urbanicity and health. Absolute numbers of migrants need to be very high, however, to make this noticeable at the aggregate level. PMID- 9876360 TI - UK study of intrapartum care for low risk primigravidas: a survey of interventions. AB - STUDY OBJECTIVE: To determine the extent of intrapartum intervention received by primigravidas. DESIGN: Cross sectional survey of NHS hospitals in the UK. SETTING: One hundred and one randomly selected hospital maternity units. PARTICIPANTS: Forty consecutive primigravid women, judged to be at low risk at the start of labour, in each hospital. MAIN OUTCOME MEASURES: Seven groups of interventions or monitoring procedures were identified from the first, second, and third stages of labour: fetal monitoring, vaginal examinations, artificial rupture of membranes, augmentation of labour, pain relief, type of delivery, and episiotomy. Data were collected during 1993. MAIN RESULTS: Ninety eight hospitals took part in the study and data were collected on 3160 low risk primigravidas. Seventy four per cent of these women had continuous cardiotocography. The proportion of women having restrictive or invasive fetal monitoring showed appreciable geographical variation for both the first and second stages of labour. Using the criterion of a vaginal examination every four hours and allowing for the length of each woman's labour, 72% had more vaginal examinations than expected; there was a significant geographical variation in the number of women receiving more than five examinations. Fifty three per cent had artificial rupture of membranes; the procedure was performed over a wide range of cervical dilatations (0 cm-10 cm). Thirty eight per cent of labours were augmented, most commonly by intravenous syntocinon; the procedure showed significant geographical variation. Twenty eight per cent had a spinal block or epidural analgesia for the relief of pain; this intervention varied by geographical region only for the second stage of labour. Over one quarter of the women required instrumental delivery. Forty six per cent had an episiotomy; the frequency of this intervention varied substantially by region. There were no infant deaths. Twelve babies were recorded at birth as having a congenital anomaly. CONCLUSIONS: The rates of several interventions seem high for this low risk group and there was substantial geographical variation in the use of six interventions. Clinical trials are needed to evaluate the optimum criteria for using these interventions from which guidelines should be drawn up by local groups and the Royal College. PMID- 9876362 TI - Super Profile analysis of socioeconomic variations in coronary investigation and revascularisation rates. AB - OBJECTIVES: To investigate socioeconomic variations in the utilisation of tertiary cardiology services. DESIGN: Cross sectional ecological study, using the Super Profile classification of enumeration districts, and ischaemic heart disease standardised mortality ratios as a proxy for needs. SETTING: The former Yorkshire Regional Health Authority area in England and its seven constituent district health authority areas. SUBJECTS: Patients with a primary diagnosis of ischaemic heart disease aged > or = 25 years who underwent investigation by angiography, or treatment by coronary artery bypass grafting (CABG) or percutaneous transluminal coronary angioplasty (PTCA), as a primary procedure between April 1992 and March 1994 in an NHS hospital. MAIN RESULTS: There is an overall increasing trend in investigation and revascularisation rates from the affluent to the deprived in the region suggesting equity. However, the gradient is slight compared with the corresponding mortality gradient. Age specific analysis shows a more appropriate trend in rates for the under 65s, but a downward trend from affluence to deprivation for the elderly. Much of the regional trend is caused by very high rates in one geographically small but densely populated district that has two tertiary cardiology centres. In other districts, with higher heart disease mortality but much lower procedure rates, there is a decreasing trend from the affluent to the deprived suggesting considerable inequity. CONCLUSIONS: This study confirms wide socioeconomic variations in coronary investigation and revascularisation within the former Yorkshire Region, suggesting that in some districts need is not being met and that service utilisation is inequitable. Such inequities are over and above those that result from proximity to tertiary cardiology centres. PMID- 9876361 TI - Poor antenatal care in 20 French districts: risk factors and pregnancy outcome. AB - STUDY OBJECTIVE: Poor attendance to antenatal visits was studied to identify risk factors and to analyse the association with adverse pregnancy outcome. DESIGN: All poor attenders and a sample of good attenders were compared within three groups of women: women < 20 years, French women > or = 20 years, and foreigners > or = 20 years. SETTING: 20 French districts including 85,000 births from January to June 1993. SUBJECTS: 848 poor attenders and 759 good attenders. Poor attenders made fewer than four antenatal visits or began care during or after the sixth month. Good attenders made at least four visits and began care before the sixth month. MAIN RESULTS: 1.1% of the women were poor attenders. Risk factors for poor attendance were single status and lack of health insurance in the group under 20; young age, high parity, and single status in the French group aged over 20; and single status and lack of health insurance in the foreign group aged over 20. For poor attenders, the odds ratios for preterm delivery were 5.8 (95% CI: 3.2, 10.5) among French women and 3.3 (95% CI: 1.5, 7.4) among foreign women with health insurance. Poor attendance was not associated with poor pregnancy outcome in the group under 20, and among foreign women over 20 without health insurance, but both groups had high rates of preterm delivery and low birth weight. CONCLUSION: Lack of health insurance is an important barrier to health care during pregnancy. Poor antenatal care is an important risk factor for adverse pregnancy outcome among women who have easy access to health care services. PMID- 9876363 TI - Misclassification of coronary heart disease in mortality statistics. Evidence from the WHO-MONICA Ghent-Charleroi Study in Belgium. AB - STUDY OBJECTIVE: To validate the Belgian vital statistics for coronary heart disease (CHD) on the basis of an independent acute myocardial infarction (AMI) register, carried out as part of the WHO-MONICA project. DESIGN: Records of fatal cases of AMI in the WHO-MONICA register were individually linked to the corresponding death certificates. SETTING: Since 1983, the WHO-MONICA Collaborating Centre Ghent/Charleroi registers all fatal and non-fatal AMI in the age group 25-69 years in two geographical areas, Ghent in the northern Dutch speaking part and Charleroi in the southern French speaking part of Belgium. Registration is done according to the MONICA protocol. The official vital statistics in Belgium are published on a yearly basis. They are essentially a reflection of the "underlying" causes of death, coded according to the 9th revision of the International Classification of Diseases (ICD). The study was undertaken in the period 1983-1991. MAIN RESULTS: Out of a total of 741 (Ghent) and 934 (Charleroi) well documented MONICA fatal cases of AMI, 492 (66.4%) and 641 (68.6%), respectively, were officially labelled as CHD (ICD code 410-414); 438 (59.1%) and 385 (41.2%), respectively, were officially labelled as AMI (ICD code 410). A substantial fraction of the MONICA AMI cases--27.1% in Ghent and 38.2% in Charleroi--was coded as "other forms of CHD" (ICD 411-414) or as "other forms of heart disease" (ICD 420-429). The remaining MONICA AMI cases--13.8% in Ghent and 20.6% in Charleroi--were classified in either very aspecific (for example, atherosclerosis, ICD 440) or totally unrelated ICD codes (for example, neoplasm, ICD 140-239). CONCLUSIONS: It is concluded from the results in this paper that a substantial part of all deaths caused by CHD in Belgium are labelled with incorrect ICD codes and are therefore misclassified in the official mortality statistics for Belgium. This is partly caused by a "drainage" of cases towards less specific CHD related ICD categories. A considerable fraction, however, seems to be absolutely misclassified. PMID- 9876365 TI - The Worcester water incident, UK: bias in self reported symptoms to an emergency helpline. PMID- 9876364 TI - A systematic review of the use of computers in the management of hypertension. AB - STUDY OBJECTIVE: To assess the effect of computers and computer-based clinical decision support systems on the management of hypertension. DESIGN: Systematic review of randomised controlled trials. SETTING: Ambulatory hypertension clinics, community-based health centres, and general practices. PARTICIPANTS: 11,962 patients enrolled in seven trials retrieved from a systematic search (electronic databases, contact with authors, reference lists; no restriction on language). MAIN RESULTS: Individual trials report on a diverse population of patients (newly diagnosed or established hypertensive patients), interventions (computers used for case finding, recall and registration, feedback on quality of blood pressure control and prescribing information), and outcomes (administration, physician performance and blood pressure control). Four of five trials reported an improvement in patient administration using a computer. Two of three trials reported an improvement in physician performance using a computer. Two of six trials. reported an improvement in blood pressure control in patients using a computer. However, positive findings in two trials should be regarded cautiously because of the potential effects of cluster randomisation. CONCLUSIONS: It seems that computers have a favourable effect on the uptake and follow up of patients in hypertension management. The effect of computers on physician knowledge, recording of information, and blood pressure control in patients is less conclusive and further studies are required. PMID- 9876366 TI - Blood pressure reactions to the cold pressor test and the prediction of ischaemic heart disease: data from the Caerphilly Study. PMID- 9876367 TI - General practice deprivation payments: are rural practices disadvantaged? PMID- 9876368 TI - Who dies from hernia? PMID- 9876369 TI - Person, time, and place predictors of seat belt use in Athens, Greece. Hellenic Road Traffic Police Department. PMID- 9876370 TI - Apoptosis, matters of life and death. PMID- 9876371 TI - A review of neuroendocrine neoplasms of the larynx: update on diagnosis and treatment. AB - Neuroendocrine neoplasms of the larynx have been divided into those of epithelial or neural origin. The latter consist of paragangliomas while the epithelial origin group can be divided into the typical and atypical carcinoids and small cell neuroendocrine carcinomata, the latter consisting of the oat cell type, the intermediate cell type and the combined cell type. There are now over 500 cases of neuroendocrine neoplasms of the larynx in the literature. The diagnosis is primarily based on light microscopy, and, in some instances, it may be supported by special histochemical studies. It should be confirmed by immunocytochemical and/or ultrastructural investigation. The different biological behaviour of neuroendocrine neoplasms of the larynx makes a specific diagnosis of paramount importance, since treatment depends on diagnostic accuracy. Typical carcinoid is an extremely rare lesion. It is treated preferably by conservative surgery; elective neck dissection is not necessary because of the lack of lymph node metastases at diagnosis. Chemotherapy and/or radiotherapy have not been effective in the limited number of patients treated thus far. Prognosis is excellent with cure following surgery. Atypical carcinoid is the most frequent non-squamous carcinoma of the larynx. The mainstay of treatment is surgery. Elective neck dissection should be performed because of the high likelihood of cervical lymph node metastases. Primary radiation therapy with adjuvant chemotherapy is not indicated. The survival rate is 48 per cent at five years and 30 per cent at 10 years. Although the larynx is one of its most common extrapulmonary sites, small cell neuroendocrine carcinoma is still a rare tumour. Surgical results for this tumour have been disappointing and is reserved for cases of local relapse with no evidence of metastasis. Chemotherapy and radiotherapy currently appear to offer the least disabling and most effective forms of therapy. The two- and five-year survival rates are 16 per cent and five per cent, respectively. Paraneoplastic syndromes have occasionally been reported in association with carcinoid tumours (typical and atypical) and small cell neuroendocrine carcinoma. There have been also rare reports of an elevated neuropeptide serum level. Paraganglioma is the only laryngeal neuroendocrine neoplasm with a female preponderance (3:1). Confusion with atypical carcinoid has led to incorrect diagnosis and inappropriate classification schemes, erroneously suggesting that laryngeal paraganglioma has the potential for aggressive behaviour. Conservative surgery represents the treatment of choice; elective neck dissection is not necessary, and the prognosis is excellent. PMID- 9876372 TI - Diabetes and hearing impairment in Mexican American adults: a population-based study. AB - Reports on the relationship between diabetes and hearing loss have been controversial. The present study examined this relationship in 1,740 Mexican American adults using data from the Hispanic Health and Nutrition Examination Survey. Hearing threshold levels were obtained for each participant at the following frequencies: 500, 1000, 2000, and 4000 Hertz (Hz). The thresholds from the worse hearing ear were used in the analyses. Diabetes and insulin use were assessed by self-report. The mean crude hearing thresholds in diabetics were significantly higher than in non-diabetics at each of the four frequencies. However, after adjustment for age, gender, and socioeconomic status, diabetics had a significantly higher mean threshold than non-diabetics only at 500 Hz (mean difference +/- SE: 2.8 +/- 1.2, p = 0.04). Diabetics who were not using insulin had significantly higher thresholds than those who were using insulin at 2000 (mean difference +/- SE: 5.6 +/- 2.6, p = 0.03) and 4000 Hz (7.7 +/- 3.3, p = 0.02). Also, at 4000 Hz, insulin users had a significantly lower mean threshold than non-diabetics (mean difference +/- SE: -4.9 +/- 1.6, p = 0.02). Our data suggest that associations between diabetes and decreased hearing acuity in the higher frequencies are present only amongst diabetic Mexican-Americans who do not use insulin. PMID- 9876373 TI - Mucoceles of the paranasal sinuses: uncommon location. AB - Mucoceles are the most common lesions causing expansion of the paranasal sinuses. The sinuses most commonly involved are, in decreasing order of frequency, frontal sinus, ethmoid sinuses, maxillary sinus and sphenoid sinus. We reviewed 46 cases of surgically proven mucoceles and the purpose of this study was to report five cases of mucoceles in an uncommon location. PMID- 9876375 TI - Primary study in experimental antiangiogenic therapy of nasopharyngeal carcinoma with AGM-1470 (TNP-470). AB - OBJECTIVE: To evaluate the efficacy of the angiogenesis inhibitor AGM-1470 for the experimental treatment of nasopharyngeal carcinoma (NPC). METHODS: A NPC human tumour model was built by tumour-bearing nude mice using the NPC cell line CNE-2. Twenty-one BALB/c nude mice bearing CNE-2 xenografts were randomized into a treatment group and a control group. In the treatment group, AGM-1470 was injected 30 mg/kg subcutaneously every other day; while the vehicle (three per cent ethanol solution in 0.9 per cent saline) was given to the mice in control group. Tumour volumes and animal weights were measured every third day. Autopsy was performed after 18 days of treatment. The tumour tissue as well as the murine tissues of heart, kidney, and liver in each mouse were removed for formalin fixation and routine HE staining. Pathological evaluation was performed in these tissues. RESULTS: There was a significant difference in tumour volume between the two groups at day 9 of treatment and this increased thereafter. At day 15 of treatment, the tumour volume was 4251 +/- 559 mm3 (n = 10) in the control group versus 3122 +/- 967 mm3 (n = 11) in the AGM-1470 treated group (p = 0.004); and T:C ratio (mean tumour volume of treated/mean tumour volume of control) was 0.73, resulting in a 27 per cent decrease in tumour growth. Central necrosis and consequential shrinkage of tumours occurred in both groups at the end of experiment. Physical toxicity and histological toxicity of heart, liver, and kidney did not result from AGM-1470 therapy. CONCLUSIONS: AGM-1470 suppresses the growth of the human NPC cell line CNE-2. Treatment by AGM-1470 has no physical nor histological toxicity. Angiogenesis inhibitors may be effective in the treatment of the local lesion of NPC. PMID- 9876376 TI - On-line resources for otolaryngologists. AB - Advances in telecommunications technology in the last decade have fostered the development of computer networks that allow access to vast amounts of information and services. The most prominent is the Internet (Glowniak, 1995). Medical information is increasingly available on such computer networks. The purpose of the present article is to provide an update to previously published otolaryngology sites (Johns, 1996; Burton and Johns, 1996) available on the World Wide Web, and to provide the otolaryngologist with details of resources that are accessible via the Internet. However, the reader should also be aware that the uniform resource locator (URL) addresses of Web sites can change without warning. PMID- 9876374 TI - Bacteriology of the maxillary and ethmoid sinuses in chronic sinusitis. AB - The bacteriology of chronic sinusitis was studied by using swab and mucosal specimens from both the maxillary and ethmoid sinuses. The specimens of the maxillary sinus were taken through translabial antroscopy. The specimens of the ethmoid sinus were taken after removing the ethmoid bulla during functional endoscopic sinus surgery (FESS). Eighty-six samples of each type of specimen were collected. Among the maxillary sinus samples, the culture rate was 60.5 per cent from the swab specimens and 36 per cent from the mucosal specimens. Among the ethmoid sinus samples, the culture rate was 58.1 per cent from the swab specimens and 75.6 per cent from the mucosal. The p-value by the Chi-Square test is higher than 0.01 (p = 0.015). As there were more isolates of Staphylococcus epidermidis from the mucosal specimens, they are not a better choice of specimen for sampling the ethmoid sinus than a swab specimen. PMID- 9876378 TI - Needle myringotomy: a useful technique with the increased prevalence of the human immunodeficiency virus. AB - Myringotomy is one of the most common procedures performed by ENT surgeons. Use of the tip of a hypodermic needle not only provides a neat and precise incision into the tympanic membrane, but is also a cheap and disposable means of doing so. Sterility is virtually guaranteed avoiding any possibility of cross contamination between consecutive patients. PMID- 9876377 TI - Audit of regional screening guidelines for vestibular schwannoma. AB - An audit of 334 patients who underwent magnetic resonance imaging (MRI) as investigation for a possible diagnosis of vestibular schwannoma was carried out to assess adherence to previously agreed screening guidelines. This represents one year's activity. A posterior fossa tumour was identified in 12 patients. Scan requests were judged to be inappropriate for 28 cases. The issues surrounding the screening for acoustic neuroma are discussed. PMID- 9876380 TI - Pyoderma gangrenosum producing saddle nose deformity. AB - Pyoderma gangrenosum affecting the nose is rare and this may lead to diagnostic confusion because of the large differential diagnosis. As diagnosis is made, largely, on the basis of exclusion the treatment of pyoderma gangrenosum may be unduly delayed. The condition is often disfiguring, particularly following inappropriate surgical intervention, and early diagnosis is therefore important. We present a case of pyoderma gangrenosum managed initially in the community with minor surgery and resulting in the rare complication of saddle nose deformity. PMID- 9876379 TI - Simultaneous leukaemic infiltrate and cholesteatoma in the same temporal bone. AB - Leukaemic relapse in the ear is very rare. We report a case of relapse of acute lymphoblastic leukaemia in the left ear. The patient presented with an aural polyp and ipsilateral facial palsy. The temporal bone showed simultaneous leukaemic infiltrate and cholesteatoma. To our knowledge, this is the first case reported with such pathology. PMID- 9876381 TI - Cavernous haemangioma in the infraorbital nerve canal--an unusual expansion in the maxillary sinus. AB - A patient with a four-week history of hypaesthesia and recurrent swelling of the cheek in the left infraorbital region presented with a computed tomography (CT) scan demonstrating an expansion in the infraorbital nerve canal, thought to be a neuroma. The tumour was surgically removed without complications and no bleeding. Histopathological investigation showed a partially thrombosed cavernous haemangioma. To our knowledge, after reviewing the literature, this is the first report of a cavernous haemangioma in the infraorbital nerve canal. PMID- 9876382 TI - Hydralazine-induced lupus and vocal fold paralysis. AB - Systemic lupus erythematosus (SLE) is a multi-system autoimmune disorder that can affect the upper airway. Hydralazine has been known to cause a lupus-like syndrome that can produce the clinical manifestations of SLE. We discuss a case of hydralazine-induced lupus, presenting with acute laryngeal oedema and right vocal fold paralysis. Cessation of hydralazine therapy resulted in reversal of paralysis. PMID- 9876383 TI - A precricoid swelling in a patient treated with Teflon injection in the vocal fold after idiopathic left vocal fold palsy. AB - Vocal fold palsy is a cause of dysphonia. Due to incomplete glottic closure during phonation, patients with a unilateral vocal fold palsy present with a weak and breathy voice and recurrent aspiration. To lessen the clinical manifestations of unilateral vocal fold palsy, polytetrafluoroethylene (Teflon) paste is one agent which has been injected into the paraglottic region, thus causing the vocal fold to move more medially. One of the complications associated with Teflon paste injection is migration of the paste into the surrounding tissues. We present a patient with idiopathic left vocal fold palsy who underwent Teflon injection to the vocal fold and subsequently developed a precricoid nodule, mimicking a cartilaginous swelling. PMID- 9876384 TI - Well-differentiated liposarcoma of the hypopharynx. AB - We present a case of well-differentiated liposarcoma of the hypopharynx in a 51 year-old male, with a review of the literature of this very rare condition. PMID- 9876386 TI - Primary presentation of spontaneous jugular vein thrombosis to the otolaryngologist--in three different pathologies. AB - An otolaryngologist is conversant with the potential of middle-ear disease leading to lateral sinus thrombosis and subsequent jugular vein thrombosis, although this is becoming an increasingly rare event. Spontaneous jugular vein thrombosis from non-otological causes, however, is not well described in the otolaryngology literature. We present three cases of jugular vein thrombosis who presented primarily to the Otolaryngology department each attributable to a different pathology including discoid lupus erythematosus, antiphospholipid syndrome and metastatic malignancy. Awareness needs to be increased of this condition in the differential diagnosis of acute neck swelling and neck pain. PMID- 9876385 TI - Malignant schwannoma of the parapharyngeal space. AB - We present a case of malignant schwannoma (MS) of the parapharyngeal space which completely encircled the internal carotid artery in close proximity to the skull base and required resection of the internal carotid artery along with the excision of the tumour. There have been three previous case reports in the literature of malignant schwannoma of the parapharyngeal space of which one case was associated with neurofibromatosis. Our patient did not exhibit any feature of neurofibromatosis. PMID- 9876387 TI - Neck abscess: an unusual presentation of a thoracic malignancy. AB - In general, malignancies presenting as abscesses are uncommon. We report on a patient that presented with a neck abscess originating in a cluster of metastatic lymph nodes from a primary squamous cell carcinoma arising in the thoracic oesophagus. PMID- 9876388 TI - Bronchogenic cyst: an unusual cause of lump in the neck. AB - Bronchogenic cysts are rare congenital benign lesions that are usually detected in the paediatric patient with symptoms of infection or compression on vital structures. They are rarely diagnosed in the adult population. We present a case of bronchogenic cyst presenting as a lump in the neck in an adult patient. Radiological imaging helped to diagnose this lesion accurately before any form of intervention. Complete surgical excision is the treatment of choice. PMID- 9876390 TI - Kikuchi's disease of the neck (histiocytic necrotizing lymphadenitis). AB - Histocytic necrotizing lymphadenitis (HNL), also known as Kikuchi's disease is a benign self limiting disease of unknown origin. It effects cervical lymph nodes, predominantly in young females. It is well described in the pathology literature, but little is written about this in the ENT literature, as few clinicians are probably aware of this condition. Five cases of cervical Kikuchi's disease are presented with a brief review of the literature. PMID- 9876389 TI - Lipomatosis of the minor salivary glands. AB - Lipomatosis has not previously been reported in minor salivary glands. Its occurrence in the parotid gland is well recognized. We present the first reported case of lipomatosis of the minor salivary glands in the nasal cavity. We also review the tumours of the minor salivary glands, lipomas and lipomatosis of the parotid, and the few reported cases of lipomas of the sinonasal tract. PMID- 9876391 TI - A visit to the Head and Neck surgery, Cancer Hospital, Beijing. PMID- 9876392 TI - Cartilage damaging activities of fibronectin fragments derived from cartilage and synovial fluid. AB - OBJECTIVE: To investigate whether fibronectin fragments (Fn-fs), shown to damage cultured cartilage, can be found in cartilage from patients with osteoarthritis (OA) or rheumatoid arthritis, or can be generated from fibronectin (Fn) within synovial fluids or from Fn in the matrix of cultured cartilage. To also determine whether cartilage or synovial fluid Fn-fs are active and, thus, could contribute to cartilage damage in vivo. METHODS: Fn-fs were immunochemically identified in cartilage extracts from patients with OA or rheumatoid arthritis or in bovine cartilage cultured with IL-1 alpha or in bovine synovial fluids treated with stromelysin-1 (MMP-3). The effect of removal of Fn-fs from OA synovial fluids was tested by passing fluids over an anti-Fn column and adding the resultant fluids to bovine cartilage cultures to measure proteoglycan (PG) degradation. Gelatin Sepharose purified Fns from bovine plasma, synovial fluid or cartilage were digested with MMP-3 and the Fn-fs tested for degradation of PG in cultured cartilage. RESULTS: Extracts of cartilage from patients with rheumatoid arthritis or with OA contained a range of Fn-fs. Removal of Fn-fs from OA synovial fluids significantly reduced the resultant damage when the fluids were added to cultured cartilage. Addition of IL-1 alpha to cultured cartilage or of MMP-3 to synovial fluids enhanced generation of Fn-fs. Fn-fs, whether derived from bovine plasma or synovial fluid or cartilage Fns, damaged cartilage. CONCLUSIONS: These data demonstrate that although Fn-fs could be generated in vivo within synovial fluids and Fn-fs found in OA synovial fluid may contribute to cartilage damage in vivo, Fn-fs could also be generated within cartilage and amplify cartilage damage. Thus, Fn-fs may be both autocrine and paracrine regulators of cartilage metabolism. PMID- 9876393 TI - An association between an aggrecan polymorphic allele and bilateral hand osteoarthritis in elderly white men: data from the Baltimore Longitudinal Study of Aging (BLSA). AB - OBJECTIVE: The aggrecan proteoglycan is a major component of articular cartilage and supports the biomechanical function of this tissue. A variable number tandem repeat (VNTR) polymorphism has been discovered recently in a region of the human aggrecan gene that codes for the chondroitin sulfate attachment sites. We examined whether alleles of this polymorphism displayed a non-random association with bilateral hand or knee osteoarthritis (OA) in men from the Baltimore Longitudinal Study of Aging (BLSA). DESIGN: DNA was obtained from 93 Caucasian men, aged 60 and above, who had bilateral hand and standing knee radiographs read for changes of OA. The DNA was analyzed by polymerase chain reaction (PCR) and/or Southern blotting for the presence of the VNTR alleles. RESULTS: Bilateral hand OA and knee OA were present in 46 and 30% of the men respectively. The following distribution of alleles was observed: allele 33 (0.5%), 29 (2.2%), 28 (31.7%), 27 (43.0%), 26 (16.7%), 25 (3.2%), 22 (2.2%) and 19 (0.5%). This distribution was similar to that detected in a random population of individuals from a separate study. In multiple logistic regression analysis, adjusting for age and body mass index, the presence of allele 27 was associated with bilateral hand OA with an odds ratio (OR) = 3.23 (95% confidence intervals (CI): 1.24-8.41). No other alleles showed an association with bilateral hand OA and the association between allele 27 and bilateral knee OA was not statistically significant (OR = 1.14; 95% CI: 0.45-2.88). CONCLUSIONS: These data demonstrate the first association between a human aggrecan gene polymorphic allele and hand OA. This finding supports the concept that genetic factors may play a role in the development and/or progression of some forms of age-onset OA. PMID- 9876394 TI - Quantitative evaluation of joint space width in femorotibial osteoarthritis: comparison of three radiographic views. AB - OBJECTIVE: Quantitative evaluation of radiographic methods proposed to improve the detection of joint space narrowing (JSN) in femorotibial osteoarthritis (OA). METHODS: Thirty-two consecutive patients with knee OA and five normal controls had three different weight-bearing radiographs of the knee: (1) anteroposterior film of both knees in full extension (extended knees), (2) anteroposterior film of one knee in extension while the patient was standing on the homolateral foot (standing on homolateral foot), (3) posteroanterior film of both knees flexed at 30 degrees (schuss view). Joint space was analyzed blind using both an evaluation of JSN with a six-grade scale (JSN score) and an image analyser computer measurement of the mean joint space width (mean JSW). The medial compartment of medial femorotibial OA knees, the lateral compartment of lateral femorotibial OA knees, as well as both compartments of control knees, were measured. Extended knee and schuss views were made 1 year later in 10 patients for the evaluation of sensitivity to change. RESULTS: The JSN scores +/- S.D. in schuss, standing on the homolateral foot and extended knee views were 2.75 +/- 1.31, 1.95 +/- 1.3 and 1.66 +/- 1.27, respectively. The mean JSW +/- S.D. in schuss, standing on the homolateral foot, and extended knee views were 2.9 +/- 1.9 mm, 3.5 +/- 1.6 mm and 3.8 +/- 1.5 mm, respectively. Changes in JSN scores and mean JSW with schuss view increased with OA severity. In controls, JSW of the medial compartment did not vary in the three views. JSW of the lateral compartment of controls was significantly larger in the schuss view. The change in JSW after 1 year was -0.41 mm (P = 0.02) in the schuss view and -0.17 mm (P > 0.05) in the extended knee view. CONCLUSION: The schuss view is suggested as the most accurate method for the evaluation of JSW in femorotibial OA. PMID- 9876395 TI - Continuous cyclic load reduces proteoglycan release from articular cartilage. AB - OBJECTIVE: To study the effect of a continuous cyclic mechanical load on the release of newly synthesized proteoglycans (PGs) from mature bovine articular cartilage. METHODS: Viable cartilage explants were continuously loaded with 1 MPa cyclic stress at 1 Hz frequency for 24 h, and the release of labeled (35SO4) PGs measured before, during and after application of the compressive load. To separate the effect of active chondrocyte catabolism from that of passive PG release, PG release in live explants, with and without protease inhibitors to inhibit PG breakdown, was compared to PG release in explants whose chondrocytes were killed prior to loading. RESULTS: In live explants, a continuous cyclic load significantly reduced PG release by as much as 50% compared to unloaded explants. In killed explants which were unloaded, the PG release increased five to 10 times, while a cyclic load reduced PG release to that found in viable, loaded explants. Twenty-four hours after load removal PG release in all loaded explants returned (increased) to that of the unloaded explants. CONCLUSIONS: These results indicate that PG release from the cartilage matrix is inhibited by continuous cyclic mechanical loading, independent of cellular metabolism, and suggest that a primary mechanism for reducing PG release is by decreasing the interstitial porosity through which the PGs can escape. PMID- 9876396 TI - Regulation of nitric oxide production by salicylates and tenidap in human OA affected cartilage, rat chondrosarcomas and bovine chondrocytes. AB - OBJECTIVE: To examine the effects of non-steroidal anti-inflammatory drugs (NSAIDS) on nitric oxide (NO) and prostaglandin E2 (PGE2) production in chondrocytes from three different species. METHODS: We have estimated NO production by Griess method, and PGE2 by RIA from the supernatants of articular cartilage obtained from osteoarthritis joints (OA-affected cartilage), rat chondrosarcomas (in ex vivo conditions) and bovine chondrocytes (stimulated with cytokines + endotoxin in vitro conditions) in the presence or absence of aspirin, indomethacin, sodium salicylate, tenidap and glucocorticoids. RESULTS: NO, which was spontaneously released in ex vivo conditions by OA-affected cartilage and rat chondrosarcomas (maintained in vivo), was susceptible to inhibition by pharmacologically relevant concentrations of aspirin, sodium salicylate and tenidap, but not to concentrations of indomethacin or glucocorticoids that significantly inhibited PGE2 production under the same conditions. Similarly, the production of NO by bovine chondrocytes grown in monolayer cultures that had been stimulated with cytokines + endotoxins (in vitro) to release both NO and PGE2 (at 48-72 h post stimulation), were inhibited by aspirin, sodium salicylate and tenidap, but not by indomethacin or glucocorticoids at concentrations sufficient to PGE2 production. Inhibition of NO in the cytokines + endotoxin stimulated bovine chondrocytes (like the human OA-affected cartilage) augmented PGE2 production. CONCLUSION: These experiments demonstrate that NO production by chondrocytes across species show a similar profile of susceptibility to inhibition by selected anti-inflammatory drugs. The insensitivity of NO production to glucocorticoids is an important characteristics of these cells that merits further investigation. PMID- 9876397 TI - Light and electron microscopic in-situ hybridization of collagen type I and type II mRNA in the fibrocartilaginous tissue of late-stage osteoarthritis. AB - OBJECTIVE: Biochemical analysis indicates the presence of collagen type I in fibrocartilaginous tissue of osteoarthritic cartilage, whereas normal hyaline cartilage contains only collagen type II produced by normal chondrocytes. Fibrocartilaginous tissue of late-stage osteoarthritis also exhibits irregularly shaped type 2b secretory chondrocytes as described in the literature. We have attempted to elucidate the type of cell which produces each type of collagen in late-stage osteoarthritis. DESIGN: We carried out in-situ hybridization at the light and electron microscopic level on the same tissue embedded in LR-Gold applying silver enhancement for gold-coupled anti-DIG antibodies. We correlated the types of cells with the expression of transcripts for type I and type II collagen. RESULTS: We found that cells resembling type 2b secretory chondrocytes of deep zones of fibrocartilaginous tissue expressed collagen type I mRNA and almost no collagen type II mRNA. The amount of collagen type I mRNA was as high as the amount produced in normal human skin fibroblasts. CONCLUSION: Some of the collagen type I in osteoarthritic human cartilage of late-stage disease is produced by cells resembling type 2b secretory chondrocytes of the deep zone. PMID- 9876398 TI - Increased secretion and activity of matrix metalloproteinase-3 in synovial tissues and chondrocytes from experimental osteoarthritis. AB - OBJECTIVE: The aim of this study was to define the relative regulation of matrix metalloproteinase-3 (MMP-3), and tissue inhibitor of metalloproteinases-1 (TIMP 1), in chondrocytes and synovium in experimental osteoarthritis (EOA). METHODS: Partial-meniscectomized (PM) rabbits, surgical sham controls (SH), and normal non surgical controls (N) were killed at times corresponding to early degenerative lesions (4 weeks) and increasingly progressive stages of EOA at 8 and 12 weeks post-PM. MMP-3 activity was measured in conditioned media from chondrocytes and synovium using a peptide cleavage assay with substance P (SP) as the substrate. TIMP-1 was quantitated using an enzyme-linked immunosorbent assay (ELISA). RESULTS: Early degenerative lesions (4 weeks post-PM) were characterized by inflammatory responses in the synovium accompanied by a significant rise of MMP-3 activity in synovial cultures (P < 0.05). At 8 weeks there was no discernible inflammation, and MMP-3 activity in EOA synovial cultures was comparable to that in the controls; this was followed by a second increase in MMP-3 activity in EOA samples at 12 weeks. MMP-3 activity was significantly elevated in EOA chondrocyte cultures at 8 weeks post-PM relative to N controls, corresponding to the most destructive phase of EOA, but not in the early phase (4 weeks) or 'late' degenerative phase (12 weeks). Medium derived from chondrocytes contained little or no TIMP-1. Synovia secreted relatively higher amounts of TIMP-1, and this was elevated at 8 weeks post-PM relative to the SH controls. The majority (approximately 90%) of MMP-3 activity could be inhibited using recombinant TIMP-1 or a hydroxamate MMP inhibitor. Complete inhibition was achieved with EDTA or 1,10 phenanthroline. CONCLUSION: Together, these data indicate that in EOA, MMP-3 is initially upregulated in the synovium which may play a pivotal role in the pathogenesis of cartilage lesions. In contrast, chondrocyte-derived MMP-3 is upregulated in the later phases of EOA, contributing further to progression of cartilage lesions. PMID- 9876399 TI - Do sunscreens prevent skin cancer? PMID- 9876400 TI - Management soon after a stroke. PMID- 9876401 TI - Glyceryl trinitrate for anal fissure? PMID- 9876402 TI - Helicobacter pylori and gastric cancer. PMID- 9876403 TI - Donepezil update. PMID- 9876404 TI - Meloxicam--a safer NSAID? PMID- 9876405 TI - Medical illustrators are required to be multi-skilled. PMID- 9876406 TI - The medical film 1897-1997: Part II. The second half-century. AB - Following the reluctance of teachers to use film for undergraduate and postgraduate medical education up until the Second World War, the active promotion of medical films after the war was encouraged by the production of catalogues of films available and the publication of medical film reviews both in journals and in books. This resulted in an increased use of films in the three decades following the war until videorecordings began to take their place and film projection is now a rarity in centres of medical education. PMID- 9876408 TI - Photography at the Department of Conservation, National Museum of Denmark. AB - The department's three photographers undertake photography to document the work of the department, both in the studio and on location. Applied photographic techniques are employed (ultra-violet, infra-red, macrophotography) and photographic processing for archival permanence is adapted to high volume monochrome machine printing. PMID- 9876407 TI - 40 years of the British Medical Association film competition. AB - Since its inception in 1957 the BMA's annual film competition has fulfilled three roles--promoting the effective use of film and video in medical education, encouraging the production of high quality audiovisual material, and helping to increase the holdings of its film library. The BMA awards recognise programmes for their clinical accuracy, educational value and creative use of the medium. PMID- 9876409 TI - John Hull Grundy, MBE, FRES, FRSA 1907-1984. PMID- 9876410 TI - Love's labour lost. PMID- 9876411 TI - Absence attributed to incapacity and occupational disease/accidents among female and male workers in the fish-processing industry. AB - Sick-leave between 1984 and 1989 was higher among both female (n = 515) and male (n = 304) fish-processing workers [observed/expected (O/E) 2.24 and 1.69, respectively] than among non-exposed groups (0.62 and 0.89). Diagnoses in the musculoskeletal system dominated (i.e., neck/upper limbs; females, exposed vs. non-exposed workers: 30 vs. 12%; males: 11 vs. 5.8%). In subjects who left employment, the O/E-ratio decreased (females: 3.02 vs. 1.55; males: 2.40 vs. 1.55). Among those women hired before the start of the observation period, exposed subjects had higher frequencies of sick-leave than non-exposed, for both total illness and musculoskeletal diagnoses. In the men, there were corresponding differences, though not fully statistically significant. Reported occupational diseases [O/E: females: 4.5; (95% confidence interval) CI = 3.2-6.1; males: 2.3; CI = 1.3-3.9] and accidents (females: 4.3; CI = 3.0-5.9; males: 1.8; CI = 1.2 2.7) were also higher in female than in male fish-processing workers, and much higher than in non-exposed workers. In conclusion, work in the fish-processing industry was associated with increased frequencies of sick-leave, especially because of diagnoses of the musculoskeletal system, and occupational disorders and accidents, in particular among female workers. PMID- 9876412 TI - Self-reported health problems among Swedish miners one year after unemployment. AB - Unemployment is considered to be a public health concern since deterioration in the health of the unemployed is often anticipated. However, for some groups, such as miners, unemployment might improve health due to a cessation of potentially harmful occupational exposures. This study evaluates the health of 79 miners in one Swedish iron-ore mine, and 226 age-matched controls from the general population, during one year after the closure of the mine. The participants received a questionnaire regarding medical history and subjective symptoms at the beginning of the study period, and after one year. Statistically significant negative effects on self-reported health attributable to unemployment were not found, although neuropsychiatric symptoms were more common among the unemployed miners. The miners reported a statistically significant improvement in grip force (p = 0.031). They had a significantly higher prevalence of symptoms associated with mining related exposures when compared with the population controls; pain in the upper extremities [relative risk (RR) = 2.27, 95% confidence interval (CI) = 1.44-3.59), back pain (RR = 1.84; CI = 1.23-2.75), vasospastic disease of the fingers (RR = 2.05; CI = 1.18-3.57) and obstructive respiratory symptoms (attacks of dyspnea and wheezing: RR = 3.67; CI = 1.16-11.6). PMID- 9876413 TI - The intention to utilize occupational health services. AB - Workers' intention to utilize the Occupational Health Service (OHS), conceived as a cost-benefit assessment of an action, is described for a series of conceivable situations. Data were acquired during interviews with a sample of 313 employees with an over-representation of workers with work-related health problems in three different companies. Only for problems that are perceived as medical, individual and work-related, do a substantial number of workers intend to utilize the occupational physician. For health and work-related problems of a collective character, the line of supervision is mostly preferred for use as an adviser. Workers' intention to utilize OHS is positively correlated with their attitude towards the occupational physician. No associations were found with self-reported health status, working environment or actual utilization of the OHS. It is concluded that the intention to utilize the OHS is an independent factor affecting the actual utilization and it should be seriously taken into consideration when evaluating or implementing the coverage by the OHS. PMID- 9876414 TI - A cross-sectional study of self-reported work conditions and psychiatric health in native Swedes and immigrants. AB - Several studies have demonstrated that immigrants in Scandinavian countries are more affected by psychosocial disabilities than the native-born population. The aim of the study was to evaluate the possible impact of work-related stressors on psychiatric health in immigrants compared to native Swedes. The study included a cluster selected cohort of 1,040 men born in 1944 (participation ratio = 79.9%), living in Gothenburg, Sweden. Of these, 182 (18.0%) were immigrants, defined as being born outside Sweden. Information on work conditions and psychiatric health were obtained by self-administered questionnaires. Employment in native Swedes showed inverse associations to frequent use of anxiolytics [relative risk (RR) = 0.2; 95% confidence interval (CI) = 0.06-0.4], frequent use of hypnotics (RR = 0.1; CI = 0.02-0.2) and use of antidepressants (RR = 0.3; CI = 0.2-0.5). None of the employed immigrants used anxiolytics or hypnotics frequently. Swedes seemed to display a number of psychiatric ill-health factors related to working conditions. These factors included frequent use of hypnotics, frequent insomnia, use of antidepressants, a high degree of melancholy, and were related to shift work, dissatisfaction with current work and management and a low degree of influence on work situation, often related to a high degree of stress at work and a frequent desire to change type of work. These associations were not seen in immigrants, apart from the risk of frequent insomnia (RR = 4.7; CI = 1.2-18.3) and dissatisfaction with colleagues (RR = 10.4; CI = 2.2-48.8) when working in shift. With a few exceptions, non-optimal working environment was associated with a low degree of life satisfaction in both groups. It was hypothesized that optimal working conditions are important for maintaining psychiatric health, and that immigrants, when employed, seem less affected by impaired working conditions than native Swedes. PMID- 9876415 TI - Smoking as a predictor of long-term work disability in physically active and inactive people. AB - Data from a Norwegian 4-year prospective questionnaire study were used to test our hypothesis that smoking is a weaker predictor of long-term work disability in physically active than inactive persons. Of 1,788 respondents who were working and not older than 62 years in 1990, 1,426 (80%) also returned a questionnaire in 1994. For respondents who were taking physical exercise less than once a week, smoking predicted long-term work disability four years later [odds ratio (OR) = 2.24; 95% confidence interval (CI) = 1.30-3.87; p < 0.01], when adjusting for age, sex, work-hours per week, heavy lifting at work, emotional symptoms and musculoskeletal pain at time 0. In respondents who were exercising at least once a week, there was no association between smoking and long-term work disability. PMID- 9876417 TI - Life events, mood, mental strain and cardiovascular risk factors in Swedish middle-aged men. Data from the Swedish part of the Renault/Volvo Coeur Study. AB - The associations between life events, mood, mental strain and cardiovascular risk factors were investigated in the Renault/Volvo Coeur Study. About 1,000 men, blue collar and white-collar workers, were asked by means of interview-administered questionnaires about life events experienced during the year preceding the screening, about mood and mental strain and about smoking, alcohol consumption and exercise habits. Blood pressure, concentration of serum lipids and blood glucose, and anthropometric measures were determined in a screening procedure. Negative life events, especially work-related, were associated with depressed mood and mental strain but not with elevation of biological risk factors such as elevated blood pressure and serum lipids. Depressed mood and mental strain were related to increased tobacco consumption in blue-collar workers and increased alcohol consumption in white-collar workers. PMID- 9876416 TI - Respiratory symptoms and sensitization in bread and cake bakers. AB - This purpose of this study was to examine the relationship between exposure to wheat flour, soya flour and fungal amylase and the development of work-related symptoms and sensitization in bread and cake bakery employees who have regular exposure to these substances. The study populations consisted of 394 bread bakery workers and 77 cake bakery workers whose normal jobs involved the sieving, weighing and mixing of ingredients. The groups were interviewed with the aim of identifying the prevalence, nature and pattern of any work-related respiratory symptoms. They were also skin-prick tested against the common bakery sensitizing agents, i.e., wheat flour, soya flour, rice flour and fungal amylase. The results of personal sampling for sieving, weighing and mixing operations at the bakeries from which the study groups were taken were collated in order to determine typical exposures to total inhalable dust from the ingredients, expressed as 8 hour time-weighted average exposures. Data from the health surveillance and collated dust measurements were compared with the aim of establishing an exposure response relationship for sensitization. The prevalence of work-related symptoms in bread bakery and cake bakery ingredient handlers was 20.4% and 10.4% respectively. However, in a large proportion of those reporting symptoms in connection with work, the symptoms were intermittent and of short duration. It is considered that the aetiology of such symptoms is likely to be due to a non specific irritant effect of high total dust levels, rather than allergy. None of the cake bakers and only 3.1% of the bread bakers had symptoms which were thought to be due to allergy to baking ingredients. Using skin-prick testing as a marker of sensitization, the prevalence of positive tests to wheat flour was 6% for the bread bakers and 3% for the cake bakers. Comparable prevalences for soya flour were 7% and 1% respectively. However, the prevalence of positive skin-prick tests to fungal amylase was 16% amongst the bread baking group with only a single employee (1%) in the cake baking group having a positive test. Furthermore, this employee had previously worked in a bread bakery. The difference in rates of sensitization to wheat flour between the bread and cake bakers is not statistically significant, whereas the difference for soya flour is at the borderline of statistical significance (p = 0.045). In contrast, the difference in fungal amylase sensitization is significant at the 0.1% level. For both bread and cake bakers, the 8 hour time-weighted average exposures for each of the activities showed a wide variation with mixing having the lowest average exposure and sieving the highest. Out of the allergens studied in this investigation, fungal amylase is the principal sensitizer in large scale bread bakeries, with the main source of exposure being the handling of bread improvers. In contrast, the risk of sensitization to wheat flour is low in both bread and cake bakeries. The absence of positive skin-prick tests in the subgroup of cake bakery employees who regularly handle fungal-amylase-containing flour suggests that their levels of exposure are below the threshold for sensitization to amylase. PMID- 9876418 TI - Development and evaluation of the use of the Internet as an educational tool in occupational and environmental health and medicine. AB - The Internet, and specifically the World Wide Web (WWW), has an important role as a method of learning in occupational and environmental health and medicine. This paper provides a systematic overview of the demands and merits of this approach to learning in a range of higher education courses in these disciplines. Drawing on a relevant theoretical framework for understanding how students learn, it describes the design and evaluation of specific resources developed for students to learn using the WWW. The occupational and environmental health or medicine components of two undergraduate degree courses and of two postgraduate courses were reviewed to determine what learning objectives would be achievable by adapting extant conventional material, or by developing new teaching and learning resources for the WWW. Depending on the objectives, various learning resource formats were developed including descriptive, interactive (such as case study or data-based), reference and self-assessment. One WWW based tutorial consisting of an interactive resource with defined objectives, linked to constantly updated, in house information and external links, was chosen as a representative for detailed evaluation. Process evaluation was based on student feedback, and outcome evaluation on group reports submitted on completion of the tutorial. Twelve of the 13 students who completed the tutorial returned the feedback questionnaire. All but one student rated it as 'good' or 'very good', with the majority of students reporting that it was easy to follow. Open-ended comments suggested that students valued the flexibility, timeliness, efficiency and breadth of access to relevant information offered by the WWW. The outcome evaluation showed that all the main learning objectives had been achieved. This work indicates that the WWW can be a valuable learning resource for occupational and environmental health and medicine. PMID- 9876419 TI - Effects of exposure to isopropyl nitrite. PMID- 9876420 TI - [Collective health: a "new public health" or field open to new paradigms?]. AB - The present essay is an exploratory study of the historical and institutional background of the so-called "crisis in public health", aimed at identifying the new trends and perspectives for the paradigmatic transformation of the health field in the context of the current international panorama of economic and cultural globalization. First, the rhetoric of health is analysed in historical perspective, briefly considering the main elements of the discourse of the ideological movements that historically built the social field of health. Medical Police, Social Medicine and Public Health are included as representative of such movements in 19th century Western Europe. After the Flexnerian turn, these movements were followed by Preventive Medicine, Community Health, Primary Health Care and Health Promotion, which dominated the scene particularly in the second half of the 20th century. The authors also summarise recent concerted PAHO efforts to debate the theory and practice of Public Health in the Americas, vis a vis the emerging demands of the economic, political and social context of Latin American countries. In this regard, the need for a common political agenda is emphasized, with the convergence of three topics-sectorial reform, "Renovation of Health for All" and the "new public health", covering the conceptual, methodological and operative domains. Secondly, a brief systematic account of the conceptual landmarks of the Collective Health movement, as carried through in the two last decades in Latin America, is presented, focusing more particularly on its potential for building up both a domain of transdisciplinary knowledge and a universe of practices. As a field of knowledge, it contributes to the study of health-disease phenomena in populations as a social process, investigating the production and distribution of disease in society as an aspect of social reproduction, and analysing health practices as a labor process integrated into the other social practices. As a universe of practices, Collective Health focuses on its models or action guidelines four objects of intervention: policies (forms of power distribution); practices (behavior modification; culture; institutions; knowledge production; institutional, professional and relational practices); technologies (organization and regulation of productive resources and processes; bodies/environments), and instruments (means of production of interventions). Finally, it is concluded that, although not being in itself a paradigm, Collective Health, as a movement committed to the social transformation of health, presents some possibilities of articulation with new scientific paradigms capable of approaching the health-disease-care object with due regard to its historicity and complexity. PMID- 9876421 TI - [Assessment of risk due to the intake of uranium isotopes in mineral spring waters]. AB - OBJECTIVE: To complement the data of a previous research concerning the evaluation of the lifetime risk of radiation-induced cancer due to the ingestion of 226Ra, 228Ra and 222Rn in mineral spring waters from a natural highly radioactive region of Brazil. The study was performed to evaluate the lifetime risk of radiation-induced cancer due to the ingestion of 238U and 234U in the same spring waters. METHOD: It is assumed that the risk coefficient for natural U isotopes is the same as for the 226Ra-induced bone sarcomas and that the equilibrium for skeletal content is 25 times the daily ingestion of 226Ra, but 11 times the daily ingestion of long-lived uranium isotopes. Waters samples were collected seasonally over a period of one year at all the spring sites used by the local population of Aguas da Prata, S. Paulo State (Brazil). RESULTS: Concentrations ranging from 2.0 to 28.4 mBq/L and from 4.7 to 143 mBq/L were observed for 238U and 234U, respectively. Based upon the measured concentrations the lifetime risk due to the ingestion of uranium isotopes was estimated. A total of 0.3 uranium-induced cancers per 10(6) exposed persons was predicted, suggesting that chronic ingestion of uranium at the levels observed at these springs will result in an incremental increase of fatal cancers of 0.1%. CONCLUSIONS: By taking into account the uncertainties in evaluating the carcinogenic effects, it can be concluded that virtually no cancer would be expected from the ingestion of uranium in the mineral spring waters analyzed. PMID- 9876422 TI - [Estimating the prevalence of height for age deficits based on the prevalence of weight for age deficits among Brazilian children]. AB - INTRODUCTION: Anthropometry is frequently used for evaluating nutritional status of individuals and populations. In recent years, community surveys have been conducted by health professionals in various regions of Brazil with the objective of complementing the data obtained through nutritional surveillance programs. One important difficulty in conducting these assessments has been measuring height during visits to the homes of survey participants. METHODS: Thirty-eight anthropometric surveys of Brazilian children aged up to 5 years using the National Center for Health Statistics (NCHS) reference were identified. The percentage of children with a Z-score below standard deviations was used to define deficits of weight for age and height for age. RESULTS: Correlation between prevalences of height for age and weight for age deficits were examined. Due to the low prevalence of deficits in weight for height in all surveys, there was a strong correlation between weight for age and height for age at the population level. Approximately 90% of the height for age (H/A) variation was accounted for by that of weight for age (W/A). CONCLUSIONS: Using the equation, (Prevalence H/A) = 0.74 + 2.34 (Prevalence W/A) -0.03 (Prevalence W/A)2 it is possible to estimate the prevalence of height deficits on the basis of prevalence of weight deficits. These results suggest that anthropometric surveys as conducted in Brazil, in the context of health services, can be simplified by measuring weight only, instead of both weight and height. PMID- 9876423 TI - [Epidemiologic study of birth weight from birth certificates]. AB - INTRODUCTION: Based on the fact that there is evidence of an association between low birth weight and neonatal and infant morbi-mortality, the World Health Organization considered birth weight as the main isolated factor associated with infant's survival. A study was undertaken to identify risk factors associated with low birth weight, using epidemiological and demographic variables present on the Birth Certificates, the source of data for the System of Information on Newborn Infants of the Brazilian Ministry of Health, for the purpose of motivating the use and the analysis of data currently generated in hospitals. METHOD: The distribution, by birth weight, of 14,784 single hospital newborn infants from five cities of the State of S. Paulo, Brazil, in an observation period of six months (1992), was analysed. Statistical analysis was based on point estimates (average, median and standard deviation) measures of association and confidence intervals (95%) of the prevalence ratios. RESULTS AND CONCLUSIONS: The highest proportion (10.4%) of low birth weight infants (weight less than 2,500 g) occurred in Itarare, the city with the highest infant mortality rate in the State, the overall proportion being 7.5%. Significant statistical associations between low birth weight and sex (female), gestational age (less than 37 weeks), mother's age (adolescent and 35 or more years of age) and parity were detected. The use of Birth Certificates in epidemiological and public health studies is highly recommended because of their validity, great importance and high coverage. PMID- 9876424 TI - [Epidemiology of drug use during the first three months of life in a urban area of southern Brazil]. AB - INTRODUCTION: The few studies on the use of medicines in children point to excessive use, and in these studies, physicians are those mainly responsible for the prescription of medicines. In order to get to know the patterns of consumption medicines better, a study was made in children in their first three months of life, according to social biological, food pattern and use of health services of variables. METHOD: The patterns of the use of medicines during the first three months of life in 655 urban children born in Pelotas, Brazil, in 1993, were described. Information on the use of medicines was collected during a two-week period in the first and third month of life. RESULTS: The use of medicines was reported by 65% of mothers at the 1st and 69% at the 3rd month of life. Seventeen per cent of children consumed three or more different medicines during those periods. Fixed combinations of three or more components-which was taken as an indicator of the poor of medicines quality-were consumed by 14% of the children at the 1st and 19% at the 3rd months of life. At the latter age, 20% of children had used a given medicine for one month or more. At the first month follow-up, Benzalkonium Chloride + Normal Saline Solution (nasal drops), Nystatin Mixture and Dimethicone + Homotropine were the most frequently used medicines. At the third month they were, Aspirin, Benzalkonium Chloride + Normal Saline Solution and Dimethicone + Homatropine. The main reasons for taking medicines were cramps at the first month and colds at the third. At the first month follow up, children with three or more siblings used 64% less medicines than the older ones. Children who were not breast-fed at the end of the first month showed a 75% greater risk of use of medicines. Similar results were observed at the third month follow-up. Some of the medicines used were not recommendable for children. CONCLUSION: Since early age children are submitted to an intense use of medicines for almost every conceivable reason, with the risk of potential side-effects and the possible lead to medicine or other drugs addiction. PMID- 9876425 TI - [Occupational accidents in Barcelona (Spain), from 1992 to 1993]. AB - INTRODUCTION: The statistics related to labor accidents as with any other notification system ought to be the basis for programs and policies with a view to the adoption of preventive measures. In order to establish preventive norms, however, the health system needs data from researchers focussing on the dynamics of and the pitfalls revealed by specific events. Within this context the main objective of this study is to proceed with an in-depth analysis of the labor accidents verified in Barcelona (Spain) using for this purpose a descriptive statistics model to test variables such as type of accident, economic sector, economic enterprise and type of labor contract. METHOD: The data source utilized was the notification system for labor accidents with grave consequences such as death of the victim registered in Barcelona during the period 1992-1993. Labor accidents registered for male workers numbered 848. A log-linear model was applied to this data base. RESULTS AND CONCLUSIONS: The results show a positive association between traumatic accidents with the construction, traffic and services sectors. A positive association was also found between traumatic accidents and the size of the company concerved the small ones being the worse type in terms of worker's injuries. Regarding the nontraumatic accidents, the study showed a positive correlation between large-sized enterprises and type of temporary worker and the civil construction sector as compared to workers with long term work contracts within industry and services. There was some evidence, also, of a positive association between small and medium sized companies and temporary work and the occurrence of work accidents. PMID- 9876426 TI - [Access to the pediatric ambulatory service at a university hospital]. AB - INTRODUCTION: In Brazil, one can verify an imbalance between the increase in the need for health care and its supply. The consolidation of the National Health System which recommends universality and equity in care, makes this issue important in the field of health service evaluation. Two pediatric services in a university hospital, one general and the other specialized are studied and compared in terms of their clients' access. METHOD: Questionnaires were applied to 221 users of the general pediatrics outpatient departments of one of the specialties with a view to studying and comparing socioeconomic and several other variables related to the access to these and other health services. RESULTS: A high level of difficulty in the users' locomotion from local health services to the hospital was noted. Of the patients attended, 40% did not receive any kind of care before their arrival and were dependent exclusively on State-run health services. The clients of the specialty were different as regards several variables when compared to the users of the general outpatients' department. The fact that they are at a better socioeconomic level and are less dependent on State-run services brings out the social inequalities involved. CONCLUSION: Socioeconomic conditions, as well as organizational aspects of the service, are seen to be both causes and consequences of social inequality verified. PMID- 9876427 TI - [Text analysis of critical areas narrated by adolescents at an advisory service]. AB - OBJECTIVE: The characterization in the discursive space of 27 users of the "Consultorio Juvenil" (young peoples' clinic), their critical areas with regard to themes, actors, actions, values and situational perception schemes. METHOD: Qualitative study with the use of semiotic analysis. Analysis was made in three steps: description of manifest text structure; codification in the quest for isotopes and internal organization analysis. RESULTS: There is clear differentiation between themes and means by gender; girls are more fully integrated into intrafamily relationships and boys into extrafamily ones. Formal engagement is a central theme for young women, while sexuality is for young men. In women a moral situation exercises influence over ethical criteria in the making of decisions; men show an individualist ethic. Both perceive problems as external, foreign to them, as part of their fate. CONCLUSIONS: Social gender formation influences the critical areas, creating myths and behavior stereotypes. The tendency to see the problem as imposed can signify a health risk for adolescents. PMID- 9876428 TI - [Gastrointestinal human myiasis caused by Eristalis tenax]. AB - The myiasis observed in Bariloche are characterized and the probable conditions under which the infestations took place established. The larvae obtained from faeces of 2 patients were identified as Eristalis tenax (Diptera: Syrphidae) according to Hartley (1961) and Organizacion Panamericana de la Salud keys (1962). These 2 cases of human gastrointestinal myiasis were the first to be registered in Bariloche (Patagonia, Argentina) and their characteristics were similar to those described for this species in other parts of the world. The lack of specific control measures in the domestic water supply system was the most probable cause of the infestation. This event extends the distribution of E. tenax and human gastrointestinal myiasis in South America to 41 degrees 03' S. PMID- 9876429 TI - [Natural features of oviposition of Culex (Melanoconion) Group Pilosus (Diptera: Culicidae)]. AB - Eggs of Culex (Melanoconion) Grupo Pilosus were found in three oviposition traps set in the toilet of a Bus Station in Joinville city, State of Santa Catarina, Brazil, during a surveillance study of Aedes aegypti. The intrinsic characteristic of group oviposition not directly on the water was confirmed. PMID- 9876430 TI - [Delayed effects of organochlorine pesticides in man]. AB - Available information on organochlorines and the chronic effects of exposure to them are set out. Organochlorinated compounds are the most persistent pesticides and can be found in all ecosystems. Although they are generally efficient in pest control, they are also a potent environment pollutant and can provoke health problems in man. The evidences of the carcinogenic potential of organochlorines are controversial and insufficient, but they have been related to an increase in the incidence of some kinds of tumors, such as leukemia and solid tumors. Reproductive effects, due to anti-androgenic and estrogenic action, on embryonic virilization, the incidence of abortion and the frequency of prematurity, have also been observed. The accumulation of the organochlorines in the adipous tissue is positively correlated to the increase in aging and could be implicated in the development of aging diseases, such as Parkinson's disease. The effects of pesticides on human health have not yet been completely elucidated. Genotoxicity is one of the most serious of the possible harmful effects caused by these compounds and calls for special attention in view of the irreversible nature of the process and to the long latency associated with its manifestation. PMID- 9876431 TI - [Evaluation of the structures of agreement and disagreement in reliability studies]. AB - The kappa coefficient has been the measurement preferred by epidemiologists for reliability studies. Various articles have demonstrated that the use of the kappa coefficient may have some undesirable features in certain contexts. Recently, methodologies using an ordinal scale for the modelling of interobserver agreement have been developed as an alternative to kappa. To show that there is a class of log-linear statistical models that when analyzed sequentially can be used to rate the patterns of agreement and disagreement. Using data on the comparability of primary and proxy respondent reports with respect to the frequency of alcoholic consumption and its correlation to coronary diseases a nested set of log-linear models was adjusted to find the "best" model. Computed odds ratios to determine the measure of agreement were also computed. The weight kappa was equal 0.685 with 95% CI (0.638-0.732) showing a good agreement. But it does not give any information about the structure of the agreement and disagreement. Among the sequence of models analyzed, the one with the best adjustment showed an agreement estimated at 0.4454 with 95% CI (0.1300-0.7608) and an association estimated at 1.3309 with 95% CI (0.9649-1.6978). The measure tau for adjacent categories was 9.2 with 95% CI (6.0-14.2). Thus, evidence shows that the observers tended to rate many phenomena similarly. Furthermore, high (or low) ratings made by primary respondents tended to be associated with high (or low) ratings made by the proxy respondents. Log-linear models can give us a more informative and more complete analysis with respect to the rating of matched pairs of observers than that given by kappa. In conclusion, the indiscriminate use of kappa as the only agreement index must be questioned. The appendix demonstrates how to use PROC GENMOD in SAS to fit these models. PMID- 9876432 TI - Research of antigen and antibodies from retroviruses, CMV and HBV among prisoners of the penitentiary complex of the region of Campinas, SP, Brazil. AB - Some viruses of the families Retroviridae, such as Human T Lymphotropic Virus (HTLV); Herpesviridae as the Cytomegalovirus (CMV) and Hepadnaviridae such as the Hepatitis B Virus (HBV) are liable to be co-transmitted with the Human Immunodeficiency Virus (HIV). Since prisoners are exposed to several and important risk factors involved in the transmission of HIV and the above mentioned viruses, male inmates from the penitentiary complex of Campinas, SP, Brazil, including HIV+ and HIV- ones, were examined for the presence of HTLV-I and/or II antibodies; IgG and IgM anti-CMV antibodies, and the research of the superficial hepatitis B antigen (HBsAg). The presence of anti-HTLV-I and/or II was determined by the Western Blot (WB) technique, whereas IgG and IgM anti-CMV and the search of HBsAg were carried out by the Microparticle Enzyme Immunoassay (MEIA-Abbott Lab). With regard to anti-HTLV-I and/or II, 58.3% (14/24-Number of positive reactions/number of sera examined) were reactive among the anti-HIV positive sera. Conversely, only 12.5% (3/24) among the HIV- negative sera showed positive reactions to HTLV-I and/or II antibodies. When looking for IgG anti-CMV percentages of 97.7% (43/44) and 95% (38/40) were obtained for anti-HIV positive and negative sera, respectively. As to IgM anti-CMV antibodies 11.36% (5/44) and 2.5% (1/40) of reactive sera were found for anti-HIV positive and negative, respectively. The HBsAg was found in 12.8% (5/39) of the sera which were anti-HIV positive. PMID- 9876433 TI - A 3-year follow-up of a Brazilian AIDS patient with protracted diarrhea caused by Enterocytozoon bieneusi. AB - Enterocytozoon bieneusi is the most prevalent microsporidian parasite that causes gastrointestinal infection in persons with AIDS. Microsporidia are increasingly recognized as important opportunistic pathogens all over the world but in Brazil only few cases have been reported due either to the non awareness of the clinical presentation of the disease or to difficulties in the laboratory diagnosis. We report a 3-year follow-up of a Brazilian HIV-positive patient in whom microsporidial spores were detected in stools and were identified as E. bieneusi using electron microscopy and PCR. The patient presented with chronic diarrhea, CD4 T-lymphocytes count below 100/mm3 and microsporidial spores were consistently detected in stools. Albendazole was given to the patient in several occasions with transient relief of the diarrhea, which reappeared as soon as the drug was discontinued. Nevertheless, a diarrhea-free period with weight gain up to 18 Kg occurred when a combination of nucleoside and protease inhibitors was initiated as part of the antiviral treatment. PMID- 9876434 TI - Elevated alanine aminotransferase (ALT) in blood donors: an assessment of the main associated conditions and its relationship to the development of hepatitis C. AB - The determination of aminotranferases levels is very useful in the diagnosis of hepatopathies. In recent years, an elevated serum ALT level in blood donors has been associated with an increased risk of post-transfusion hepatitis (PTH). The purpose of the study was to research the factors associated with elevated ALT levels in a cohort of voluntary blood donors and to evaluate the relationship between increased ALT levels and the development of hepatitis C (HCV) infection. 166 volunteer blood donors with elevated ALT at the time of their first donation were studied. All of the donors were questioned about previous hepatopathies, exposure to hepatitis, exposure to chemicals, use of medication or drugs, sexual behaviour, contact with blood or secretions and their intake of alcohol. Every three months, the serum levels of AST, ALT, alkaline phosphatase, gamma glutamyl transpeptidase, cholesterol, triglyceride and glycemia are assessed over a two year follow-up. The serum thyroid hormone levels as well as the presence of auto antibodies were also measured. Abdominal ultrasound was performed in all patients with persistently elevated ALT or AST levels. A needle biopsy of liver was performed in 9 donors without definite diagnostic after medical investigation. The presence of anti-HCV antibodies in 116 donors were assayed again the first clinical evaluation. At the end of follow-up period (2 years later) 71 donors were tested again for the presence of anti-HCV antibodies. None of donors resulted positive for hepatitis B or hepatitis C markers during the follow-up. Of the 116 donors, 101 (87%) had persistently elevated ALT serum levels during the follow-up. Obesity and alcoholism were the principal conditions related to elevated ALT serum levels in 91/101 (90.1%) donors. Hypertriglyceridemia, hypercholesterolemia, hypothyroidism and diabetes mellitus also were associated with increased ALT levels. Only 1/101 (0.9%) had mild chronic active non A-G viral hepatitis and 3/101 (2.9%) had liver biopsy with non-specific reactive hepatitis. The determination of ALT levels was not useful to detect donors infected with HCV at donation in Brazil, including the initial seronegative anti HCV phase. PMID- 9876435 TI - Detection of IgM antibodies to Schistosoma mansoni gut-associated antigens for the study of the dynamics of schistosomiasis transmission in an endemic area with low worm burden. AB - For a period of 2 years, five follow-up measures of prevalence and incidence rates were estimated in a prospective study of S. mansoni infection in a group of schoolchildren who were living in a rural area of the Municipality of Itariri (Sao Paulo, Brazil), where schistosomiasis is transmitted by Biomphalaria tenagophila. Infection was determined by the examination of three Kato-Katz stool slides, and the parasitological findings were analyzed in comparison to serological data. In the five surveys, carried out at 6-month intervals (March April and September-October), the prevalences were, respectively, 8.6, 6.8, 9.9, 5.8 and 17.2% by the Kato-Katz, and 56.5, 52.6, 60.8, 53.5 and 70.1% by the immunofluorescence test (IFT). Geometric mean egg counts were low: 57.8, 33.0, 35.6, 47.3 and 40.9 eggs per gram of feces, respectively. Of the total of 299 schoolchildren, who submitted five blood samples at 6-month intervals, one for each survey, 40% were IFT-positive throughout the study, and 22% were IFT negative in all five surveys. Seroconversion from IFT negative to positive, indicating newly acquired S. mansoni infection, was observed more frequently in surveys carried out during March-April (after Summer holidays), than during September-October. Seasonal trends were not statistically significant for detection of S. mansoni eggs in stool. The results indicate that the use of IgM IFT is superior to parasitological methods for detection of incidence of S. mansoni infection in areas with low worm burden. PMID- 9876436 TI - Salmonella-S. mansoni association in patients with acquired immunodeficiency syndrome. AB - Two young men with Salmonella bacteraemia, active schistosomiasis and the acquired immunodeficiency syndrome are reported. The clinical presentation comprised nonspecific signs and symptoms, such as fatigue, malaise, weight loss, diarrhoea, prolonged fever, and hepatosplenomegaly. In one patient, liver biopsy showed poorly formed granulomata around Schistosoma mansoni eggs and hepatitis. Treatment of schistosomiasis alone induced consistent clinical improvement with eventual cure of both Salmonella and S. mansoni infections. Recognition of the Salmonella-S. mansoni association in patients with AIDS is important because treatment of schistosomiasis makes a difference, improving the prognosis of this otherwise, recurrent, potentially fatal bacteraemia. PMID- 9876437 TI - Intracellular Leishmania amazonensis killing induced by the guanine nucleoside 8 bromoguanosine. AB - In this study we investigated the effect of 8-Bromoguanosine, an immunostimulatory compound, on the cytotoxicity of macrophages against Leishmania amazonensis in an in vitro system. The results showed that macrophages treated with 8-Bromoguanosine before or after infection are capable to reduce parasite load, as monitored by the number of amastigotes per macrophage and the percentage of infected cells (i.e. phagocytic index). Since 8-Bromoguanosine was not directly toxic to the promastigotes, it was concluded that the ribonucleoside induced macrophage activation. Presumably, 8-Bromoguanosine primed macrophages by inducing interferon alpha and beta which ultimately led to L. amazonensis amastigote killing. The results suggest that guanine ribonucleosides may be useful to treat infections with intracellular pathogens. PMID- 9876438 TI - Pulmonary scedosporiosis. AB - A case of a solitary pulmonary nodule due to Scedosporium apiospermum (Pseudallescheria boydii) is related. A review of the pertinent literature was done and, in addition, similar lesions caused by other opportunistic fungi are commented. PMID- 9876440 TI - Ectopic cutaneous schistosomiasis: report of two cases and a review of the literature. AB - Two cases of ectopic cutaneous schistosomiasis are described. Both patients presented with abdominal papular skin lesions, which on biopsy were found to contain granulomas with Schistosoma mansoni eggs. Twenty-five other cases were retrieved from the literature. Most patients were female, mean age 24.9 year, with a predominance of the white race. The most common localization was anterior thorax and abdomen. Usually, the lesions were asymptomatic. In few cases, however, severe clinical syndromes due to the parasite coexisted, such as transverse myelitis or the acute-toxemic form of the disease. Intestinal infection was not frequently demonstrated in these patients. The importance of the recognition of these cutaneous lesions may rest on the opportunity to provide an etiological diagnosis in these difficult cases. PMID- 9876439 TI - Vertical transmission of HTLV-I/II: a review. AB - The vertical transmission of the human T-cell lymphotropic virus type I (HTLV-I) occurs predominantly through breast-feeding. Since some bottle-fed children born to carrier mothers still remain seropositive with a frequency that varies from 3.3% to 12.8%, an alternative pathway of vertical transmission must be considered. The prevalence rate of vertical transmission observed in Japan varied from 15% to 25% in different surveys. In Brazil there is no evaluation of this form of transmission until now. However, it is known that in Salvador, Bahia, 0.7% to 0.88% of pregnant women of low socio-economic class are HTLV-I carriers. Furthermore the occurrence of many cases of adult T-cell leukemia/lymphoma and of four cases of infective dermatitis in Salvador, diseases directly linked to the vertical transmission of HTLV-I, indicates the importance of this route of infection among us. Through prenatal screening for HTLV-I and the refraining from breast-feeding a reduction of approximately 80% of vertical transmission has been observed in Japan. We suggest that in Brazil serologic screening for HTLV-I infection must be done for selected groups in the prenatal care: pregnant women from endemic areas, Japanese immigrants or Japanese descendents, intravenous drug users (IDU) or women whose partners are IDU, Human immunodeficiency virus carriers, pregnant women with promiscuous sexual behavior and pregnant women that have received blood transfusions in areas where blood donors screening is not performed. There are in the literature few reports demonstrating the vertical transmission of HTLV-II. PMID- 9876441 TI - Prevalence of anti-Toxocara antibodies in a random sample of inpatients at a children's hospital in Vitoria, Espirito Santo, Brazil. AB - In the streets of Vitoria, in the State of Espirito Santo, Brazil, are large number of stray dogs, many of which are infected with Toxocara canis, suggesting a high risk for human infection. In order to investigate the prevalence of Toxocara infection in children in Espirito Santo we studied the prevalence of anti-Toxocara antibodies in 100 random inpatients over one year of age, at the Children's Hospital N.S. da Gloria, the reference children's hospital for the State. All the sera were collected during the period between October 1996 and January 1997. The mean age was 6.6 +/- 4.1 yrs. (1 to 14 yrs., median 6 yrs.) and there were patients from all of the different wards of the hospital. Sixty-eight patients came from the metropolitan area of Vitoria and the other 32 from 17 other municipalities. The anti-Toxocara antibodies were investigated by ELISA-IgG using a secretory-excretory antigen obtained from second stage larvae. All sera were adsorbed with Ascaris suum antigen before the test. Thirty-nine sera (39%) were positive, predominantly from boys, but the gender difference was not statistically significant (boys: 25/56 or 44.6%; girls: 14/44 or 31.8%; p = 0.311). The prevalence of positive sera was higher, but not statistically significant, in children from the urban periphery of metropolitan Vitoria (formed by the cities of Vitoria, Cariacica, Vila Velha, Serra and Viana) than in children from 17 other municipalities (44.1% and 28.1% respectively, p = 0.190). Although the samples studied do not represent all children living in the State of Espirito Santo, since the Children's Hospital N.S. da Gloria admits only patients from the state health system, it is probable that these results indicate a high frequency of Toxocara infection in children living in Espirito Santo. Further studies of population samples are necessary to ascertain the prevalence of Toxocara infection in our country. PMID- 9876442 TI - Thermotolerant Campylobacter species isolated from psittaciformes in the Peruvian Amazon region. PMID- 9876443 TI - Laboratory diagnosis of acute human parvovirus B19 infection by specific IgM detection. PMID- 9876444 TI - A tribute from the Tropical Medicine Institute of Sao Paulo in memory of Leonidas de Mello Deane and his wife Maria Jose von Paumgartten Deane. PMID- 9876445 TI - The Cochrane collaboration in the Third World. PMID- 9876447 TI - Preoperative irradiation therapy and radical hysterectomy: prognostic value of tumor regression after initial irradiation of squamous cell carcinoma of the cervix. AB - OBJECTIVE: To investigate the role of tumor persistence in patients submitted to irradiation therapy and radical hysterectomy. DESIGN: A retrospective analysis of prognostic factors. LOCATION: Hospital A.C. Camargo, Sao Paulo, Brazil, a private non-profitmaking foundation and tertiary referral centre. PATIENTS: A total of 629 cases of invasive squamous cell carcinoma of the cervix were studied. Criteria for inclusion in the study were: confirmed histological diagnosis of squamous cell carcinoma and no previous treatment (except for preoperative radiotherapy carried out at the Hospital A.C. Camargo itself). At the end of the follow-up period, 410 patients (65%) had no evidence of disease and 219 (34.8%) had died because of the tumor. INTERVENTION: The patients were submitted to radical surgery and radiation therapy, separately or in combination between 1953 and 1982. MAIN OUTCOMES MEASURES: Multivariate analysis of the different variables was performed according to the Cox regression method. RESULTS: The variables of prognostic value were, in decreasing order of importance: the decade of patient admission (p = 0.0001), the modality of therapy employed (p = 0.0005), the presence of residual tumor in the surgical specimens (p = 0.0055) and the clinical stage of the disease (p = 0.0575). CONCLUSION: Radiation therapy controlled a considerable number of local tumors and pelvic lymph nodes but not all of them in every patient. There is a specific group of patients for whom radical surgery is necessary to achieve control of the disease. PMID- 9876446 TI - Epidemiology of acute hepatitis B in a university hospital in Sao Paulo, Brazil: retrospective study of two five-year periods. AB - CONTEXT: HBV infection is endemic in Brazil and acute HBV infection is still a common disease. OBJECTIVE: To analyze incidence, risk factors and evolution of acute HBV infection. SETTING: University Hospital. PATIENTS: 357 patients with acute HBV infection, comparing two periods: 1985-1989 vs. 1990-1994. RESULTS: The overall incidence declined from 50 new cases/year in 1985-89 (30% of all cases) to 25 new cases/year in 1990-94 (8% of all cases). Transmission among male homosexuals (3.9% of cases in 85-89) declined to 1.3% in 90-94 (p > 0.05). Amongst health care workers (HCW) it declined from 8.2% to 2.0% (p = 0.02). Conversely, heterosexual transmission increased from 4.8% to 10.1% (p = 0.06). Chronification of HBV infection following the acute episode was observed in 1.7%. Fulminant hepatic failure was seen in 3.4%. However, 27.2% of patients were lost to follow-up before normalization of the biochemical tests. CONCLUSIONS: The different patterns of risk factors observed is probably related to measures for preventing AIDS and to HCW vaccination programs. Chronification following acute episodes was not a common event. PMID- 9876448 TI - Endolymphatic irradiation in preparation for renal transplantation: a 26-year's follow-up. AB - OBJECTIVE: The aim of the present study was to analyze the long-term evolution of patients submitted to endolymphatic irradiation as a pre-transplant preparation. SETTING: Referral center of university hospital. DESIGN: Case-control study. MAIN OUTCOMES MEASURES: The study was designed to evaluate the incidence of rejection, kidney loss, leukopenia, infection, and graft survival in the group treated (group 1) prior to surgery, compared to a control group (group 2) composed of patients under identical clinical conditions (sex, age, type of donor, immunosuppressive therapy and time of transplant) that did not undergo treatment preparation. PATIENTS: Patients were selected from amongst transplantation candidates on a long-term waiting list, some with a high level of antibodies against panel. The control group was chosen from amongst recently transplanted patients. Patients in the treated group received lipoiodine containing 131I with specific activity ranging between 4 and 6 mCu/ml. RESULTS: A significant difference between the two groups was found with regard to the incidence of rejection crises (21.0% in group 1 and 73.6% in group 2; P = 0.003), and the maintenance dose of azathioprine (smaller in group 1; P < 0.01). As to kidney graft loss due to rejection, a tendency to significance could be identified (10.5% in group 1 and 42.1% in group 2; P = 0.063); however, the difference was not significant between the two groups in terms of reversibility of rejection episodes during the first 60 post-transplant days. CONCLUSIONS: The authors concluded that this method, besides being relatively innocuous (there was no compromising of either the thyroid gland or of gonad function and there was no increase in tumor incidence), has an extended immunosuppressive effect, and can be indicated for cadaveric renal allograft recipients, especially those showing high panel reactivity. PMID- 9876449 TI - Neurologic manifestations of AIDS: a review of fifty cases in Santos, Sao Paulo, Brazil. AB - OBJECTIVE: To review the neurologic manifestations of AIDS in patients who were admitted to Hospital Guilherme Alvaro (HGA) due to any clinical manifestation of the disease. DESIGN: Case series. PATIENTS: All HIV+ patients admitted to the Faculty Hospital (HGA) between July 96 and April 97 were included in this review. RESULTS: From the 117 HIV+ patients admitted to hospitalization due to AIDS related symptoms, 50 (42.7%) presented neurologic manifestations. The most prevalent of these was neurotoxoplasmosis (68%), but a variety of other neurologic diseases were observed. Only 36% of these 50 patients had neurological signs and symptoms as the main complaint for admission, 12% of the patients had at least complained of some neurologic dysfunction at the time of admission and 10% had no neurologic complaints at all. The remaining 42% (21 patients) only complained of neurologic manifestations of AIDS when specifically asked. CONCLUSIONS: The prevalence of neurologic manifestations of AIDS is very high in patients admitted to hospital. Even in the absence of neurologic-related complaints, these patients have to be carefully questioned and examined in the search for an underlying neurologic complication which may present high morbidity and mortality. PMID- 9876450 TI - Variations in white blood count, thromboxane B2 levels and hematocrit in chronic venous hypertension. AB - OBJECTIVE: To analyze variations in leukocyte count and thromboxane B2 production in the femoral vein of patients with chronic venous hypertension (CVH). DESIGN: Prospective clinical study, controlled, non randomized and open. LOCATION: Hospital das Clinicas, Faculdade de Medicina da Universidade de Sao Paulo, referral center, university hospital. PARTICIPANTS: 15 patients with recurring stasis ulcer were analyzed, selected randomly from the venous diseases outpatient center, and 4 without lower limb venous alterations were also analyzed. INTERVENTION: Blood samples from the femoral and brachial veins were drawn following supine and 45 degrees reverse Trendelenburg. MAIN OUTCOMES MEASURES: Direct leukocyte count and analysis of the thromboxane B2 with enzyme linked immunosorbent assay test. RESULTS: After 30 minutes in reverse Trendelenburg, patients with CVH showed a leukocyte count reduced by +/- 27% (p = 0.02) and thromboxane B2 levels increased by +/- 158% (p = 0.02). CONCLUSIONS: We suggest that future studies of medications for stasis ulcers include their effects on leukocyte entrapment and thromboxane B2 production in the lower limb venous system. PMID- 9876451 TI - Oropharynx microbiota among alcoholics and non-alcoholics. AB - CONTEXT: The oropharynx microbiota plays an important role in the origin of infections, especially among alcoholics whose airway defenses are impaired. OBJECTIVE: To compare the normal oropharingeal flora in heavy alcohol drinker and non-alcoholics. PATIENTS: 117 persons, 58 heavy alcohol drinkers and 59 non alcoholics. SETTING: Santa Casa de Sao Paulo Emergency Service. DESIGN: A blind prospective study. MAIN OUTCOMES MEASURES: Prevalence of aerobic and anaerobic bacteria, and fungi. RESULTS: The study of the oropharynx microbiota among heavy alcohol drinkers demonstrated the presence of anaerobic microorganisms in 84.5% of them, including: Bacteroides sp, Prevotella melaninogenica, Fusobacterium sp, Veilonella sp, Peptostreptococcus sp, Propionibacterium sp, Bifidobacterium sp and Clostridium sp, versus 30.5% (p < 0.005) of non-alcoholics. Candida sp was present in 34.5% of heavy alcohol drinkers and 5.1% of non-alcoholics (p < 0.005). Enterobacteria predominated among heavy alcohol drinkers (25%) compared with non-alcoholics (5.5%) only in the age group 14 to 34 years (p < 0.05). CONCLUSION: Based upon these results, it was possible to conclude that the knowledge of the oropharynx microbiota among heavy drinkers and non-alcoholics has an important predictive value concerning probable etiologic agents of lower airway infections. Infections caused by anaerobic microorganisms and fungi should be taken into consideration during the choice of empirical therapy for heavy alcohol drinkers. PMID- 9876453 TI - Websites for critical appraisal in diagnosis and treatment. PMID- 9876452 TI - Correlation between endometrial dating of luteal phase days 6 and 10 of the same menstrual cycle. AB - CONTEXT: Endometrial maturation, important in the diagnosis of infertile couples, has been evaluated since 1950 using the Noyes criteria. Nevertheless, there is no consensus regarding the most suitable period of the luteal phase for performing the biopsy. OBJECTIVE: This study evaluated the correlation between the histological dating of two endometrial biopsies performed in the same menstrual cycle, on luteal phase days six and ten. DESIGN: Prospective study. SETTING: Human Reproduction Division of the Federal University of Sao Paulo, referral center. PATIENTS: Twenty-five women complaining of infertility had their menstrual cycles monitored by ultrasound and LH plasma levels, to obtain evidence of ovulation. PROCEDURES: Endometrial biopsies were performed on luteal phase days LH + 6 and LH + 10 (luteal phase day 1 = LH + 1 = the day that follows LH peak). Dating was done according to morphometric criteria, in which an endometrium sample is considered out of phase if the minimum maturation delay is one day. On day LH + 6, blood was drawn for plasma progesterone level determination. RESULTS: All patients had an ovulatory cycle (mean LH peak: 47.4 U/L; mean follicular diameter on LH peak day: 18.9 mm; mean endometrial thickness on LH peak day: 10.3 mm; mean plasma progesterone level on day LH + 6: 14.4 ng/ml). 14 patients had both biopsies in phase; 5 patients had out of phase biopsies only on day LH + 6; 3 had out of phase biopsies only on day LH + 10 and 3 patients had out of phase biopsies on both days. McNemar's test showed no statistical difference between these data (p > 33.36%). CONCLUSIONS: The correlation found between the endometrial datings suggests that biopsies performed on either of these two days are suitable for evaluation of endometrial maturation. PMID- 9876454 TI - Carcinoid tumor of the gallbladder. AB - The carcinoid tumor is a relatively rare type of endocrine tumor, which arises mainly in the gastrointestinal tract. Primary gallbladder and biliary duct system carcinoids comprise less than 1% of all carcinoid tumors arising from any tissue or organ in the body. We describe a case of carcinoid tumor of the gallbladder in a 39-year-old man. There have been only 32 cases described in the literature. PMID- 9876455 TI - [In vivo and in vitro study of the marginal sealing in ceramic-covered gold inlays]. AB - In the view of the patients, Galvano-Inlays fulfil esthetic requirements in contrast to cast restaurations. However, it was decided to evaluate the fine gold margins. The marginal seal of Galvano-Inlays in different stages of production was evaluated by means of light microscopy, and scanning electron microscopy. In addition Galvano-Inlays which had been in the mouth for two years, were evaluated in the same way. Besides good contact of the fine gold to the tooth, marginal discrepancies in the range of 60 to 160 microns were found which can hardly be improved by means of burnishing. After all, the insufficiency results from the inevitable gap of cementation or from breaking failure of the polished and thus hardened margin of the fine gold. PMID- 9876456 TI - [Relationship between dental status and salivary microbiology in adolescents]. AB - The authors studied caries prevalence in connection with salivary caries related findings, in 349, 14-16 years old Hungarian adolescents, living in two different cities. DMF.T, DMF-S means, stimulated salivary flow, buffer capacity, Streptococcus mutans, Lactobacillus and Candida albicans counts in saliva were determined. The ratio of cariesfrre adolescents was 4.6% in the total population sample, DMF-T mean values were 7.24 +/- 4.86, DMF-S means 10.50 +/- 8.35. Mean secretion rate of stimulated saliva was 0.84 +/- 0.50, a low buffer capacity was found in 6.3% of the examined children. Ratio of carriers of Streptococcus mutans, Lactobacilli and yeasts in saliva was 89.7%, 73.9% and 47.7% respectively. DMF-T and DMF-S values, as well as S. mutans counts were lower in the capital than in the other city. Statistically significant correlations were found between DMF-S, DMF mean values and salivary microbiological counts. PMID- 9876457 TI - [Evaluation of the dental status from he viewpoint of denture requirements in the adult population of Hungary]. AB - Authors found a rather high number of missing teeth of the Hungarian population, whereas the claim of the patients to dentures is less than the capacity of the dental professionals. The percentage of the patients wearing fixed and/or removable dentures is only around 50% (mean value), in the aesthetically not disadvantageous cases (i.e.: classes 1/a; 2/a; Fabian-Fejerdy) only 30-40%, but in the aesthetically disadvantageous cases 70-80% (i.e.: classes 1/b; 2/a/1; 3; Fabian-Fejerdy). The percentage of the men wearing dentures is higher than the percentage of the men (61.9% and 44.9% respectively). Authors concluded that, the most important expectations of the patients to dentures are aesthetic. The injurious effects of not reconstructed missing teeth have minor effects on the patients motivation. PMID- 9876458 TI - [Current therapy of chronic renal failure]. AB - The course of chronic renal failure is generally progressive and mediated by several factors that operate in combination. Several extrarenal events which may cause transient or permanent deterioration of renal function, are important, because their correction may slow the progression of renal disease e.g. volume disorders, infection, nephrotoxic agents. In progression of chronic renal disease leading factors are hypertension, proteinuria and high protein/phosphorus intake. Number of evidence suggests that ameliorating hypertension, reducing proteinuria slow the progression of chronic renal failure. Clinical studies in diabetic nephropathy demonstrated that the renoprotective effect of ACE inhibitors was independent of their effect of systemic blood pressure. In ESRD patients access for renal replacement therapy should be obtained as early as possible. An A-V fistula may take several weeks to mature especially in diabetic or elderly patients. Early dialysis has been advocated in diabetic patients. In general, patients can start ESRD therapy when residual kidney function drops to 5-10% of normal value. High quality of dialysis should be provided to the uremic patient with respect of successful renal transplantation. PMID- 9876459 TI - [Human herpesvirus-8 in Hungarian blood donors and in patients with Kaposi sarcoma]. AB - The pathogenic role of Kaposi's sarcoma associated herpesvirus or human herpesvirus-8 in the development of Kaposi's sarcoma is widely accepted today. Here the authors report an indirect immunofluorescence assay based seroepidemiologic study. Their results indicate that 1.56% (17/1089) of the Hungarian adult population and 100% (12/12) of Kaposi's sarcoma patients have serum antibodies to HHV-8 latent nuclear antigens. PMID- 9876460 TI - [Magnetic resonance cholangiography: our first experience]. AB - Magnetic resonance cholangiography is a new field of clinical application within MR imaging, enabling high-quality imaging on disorders of the pancreaticobiliary system. This cross-sectional imaging technique is, by means of postprocessing, suitable for the production of reconstruction images appropriate for comparison with endoscopic retrograde cholangiography. The quality of the images thus obtained is dependent upon the level of development of the technology and software available. Observations arising from authors initial examinations using 2D fast spin echo sequence are summarised. PMID- 9876461 TI - [Fetal ethology]. AB - The author summarises the main aspects of fetal activity and behavior. Fetal activity provides information about early stages of the neurodevelopmental processes. Fetal behavior offers an opportunity for the study of motor, sensory, and cognitive functions in the fetus and, thus, of fetal wellbeing. In high risk pregnancies, studies of fetal behavior may help to establish the optimal time for the delivery of the fetus. PMID- 9876462 TI - [Fibroepithelial polyp causing ureteropelvic obstruction in childhood]. AB - The fibroepithelial polyp is a benign tumour that occurs as a rare intraluminal mass within the urinary tract. Most commonly it is located within the ureter or ureteropelvic junction. The authors present 3 cases of fibroepithelial polyp causing obstruction of the ureteropelvic junction. Dismembering pyelonplasty resulted in disappearance of the hydronephrosis, and the postoperative course was uneventful. PMID- 9876463 TI - [Phrenology as an unsuccessful adventure in cerebral localization (Franz Joseph Gall: 1758-1828)]. PMID- 9876464 TI - [Immunoprophylaxis of hepatitis in the Hungarian literature]. PMID- 9876465 TI - [Results of BCG vaccination in Hungary since 1929: evaluation of its preventive and immunotherapeutic uses]. PMID- 9876466 TI - [Botulism in the domestic literature]. PMID- 9876467 TI - [Significant increase of pectus excavatum cases in Tata between 1990 and 1992]. PMID- 9876468 TI - Sensitive measures of nutritional status in children in hospital and in the field. AB - Protein-energy malnutrition and obesity are the most common nutritional disorders that complicate the clinical course of children with neoplastic diseases. Sensitive measures of nutritional status should be used to detect these problems in children with cancer. Height and weight measurements are the mainstay of the nutritional assessment of the child. These measurements can be converted to growth velocities or to height-for-age and weight-for-height Z-scores or percent of expected values to provide a measure of the degree of under- or over-nutrition in the child. Skinfold thickness and circumference measurements of the arms, legs and/or trunk may be useful to characterize the changes in peripheral fat depots and muscle mass, respectively. However, the assessments of body composition using these measurements are subject to methodological error because selected skinfold sites are excluded. Whole-body potassium, measured by 40K counting, and total body water, measured by deuterium or 18O dilution, serve as "gold standards" to determine the lean body mass and body fat status of the child, but these techniques may not be practical in all settings. The assessment of the nutritional status of the child serves as a guide to early nutritional intervention. Indicators for early nutritional intervention include: (1) height for-age and weight-for-height or -age Z-scores more than 2 SD below the mean for age, (2) height-for-age measurements less than 95% of expected, (3) weight-for height measurements less than 90% or greater than 120% of expected and (4) height velocities less than 5 cm/year after 2 years of age. Early nutritional intervention is essential to restore normal body composition, reverse linear growth arrest, promote tolerance to chemotherapeutic and radiation regimens and improve the quality of life in children with cancer. PMID- 9876469 TI - Sensitive measures of the nutritional status of children with cancer in hospital and in the field. AB - Sensitive measures of nutritional status exist. Initial assessment must include some measure that is independent of tumour mass, particularly in children with large solid tumours. Arm anthropometry, including triceps and biceps skinfold thickness (SFT), and mid-upper-arm circumference (MUAC) are ideal in this situation, but MUAC is probably the simplest measure to use. In the clinical setting, a direct measure of fat-free body mass (FFBM) does not exist, but bio electrical impedance (BIA) measures FFBM indirectly, and has many advantages, in particular its ease of use and immediate results. The BIA analyzer is portable and hence can be used in the field as well as by the bedside. Serum proteins and insulin-like growth factors are insufficiently sensitive as nutritional indices and have only a minor role in nutritional assessment. PMID- 9876470 TI - Identification of risk factors for malnutrition: is there some evidence for predisposition? AB - Reviewed are reports on factors, identified by risk analysis, involved in the genesis of primary malnutrition in children. Data are compared with the sequence of factors in a flow diagram, based on the natural history of malnutrition, proposed 3 decades ago. Susceptibility to malnutrition is analyzed in light of observations related to inheritance, the ob gene and leptin. PMID- 9876471 TI - Prenatal vitamin supplementation and risk of childhood brain tumors. AB - An international case-control study of primary pediatric brain tumors included interviews with mothers of cases diagnosed from 1976 to 1994 and mothers of population controls. Data are available on maternal vitamin use during pregnancy for 1,051 cases and for 1,919 controls from 8 geographic areas in North America, Europe and Israel. While risk estimates varied by study center, combined results suggest that maternal supplementation for 2 trimesters decreased risk of brain tumor [odds ratio (OR) = 0.7; 95% confidence interval (CI) = 0.5, 0.9], with a trend of less risk with longer duration of use (p trend = 0.0007). The greatest risk reduction was among children diagnosed under 5 years of age whose mothers used supplements during all 3 trimesters (OR = 0.5; CI = 0.3, 0.8). This effect did not vary by histology and was seen for supplementation during pregnancy rather than during the month before pregnancy or while breastfeeding. Our findings are largely driven by data from the United States, where most mothers took vitamins. The proportion of control mothers who took vitamins during pregnancy varied markedly from 3% in Israel and in France, 21% in Italy, 33% in Canada and 52% in Spain to 86-92% at the 3 U.S. centers. The composition of the various multivitamin compounds taken also varied: daily dose of vitamin C ranged from 0 to 600 mg; vitamin E from 0 to 70 mg; vitamin A from 0 to 30,000 IU; and folate from 0 to 2,000 micrograms. Mothers also took individual micronutrient supplements (e.g., vitamin C tablets), but most mothers who took these also took multivitamins, making it impossible to determine the potential independent effects of these micronutrients. PMID- 9876472 TI - Maternal diet during pregnancy and risk of brain tumors in children. AB - Nine studies of childhood brain tumors and maternal diet during pregnancy have focused on foods related to the N-nitroso-compound(NOC) hypothesis. An association between frequent consumption of cured meat by pregnant women and increased risk is a consistent finding in most of the studies. The data on fruit and vegetable consumption are less consistent, but suggest decreased risk. Studies that assess all aspects of maternal diet during pregnancy are needed to determine whether the observed associations remain after adjustment for other aspects of diet. Such comprehensive studies also may elucidate other dietary factors that affect the risk of brain tumors in children. PMID- 9876473 TI - Maternal diet and infant leukemia: a role for DNA topoisomerase II inhibitors? AB - Leukemia in the first year of life is extremely rare world-wide. However, unlike leukemias in older children, nearly 75% of infant leukemias demonstrate a specific abnormality involving a gene, MLL, on chromosome band 11q23. Molecular studies suggest strongly that these leukemias occur in utero. Treatment-related acute myeloid leukemias (AML), associated with specific chemotherapeutic agents that inhibit DNA topoisomerase II (topo 2), also manifest identical abnormalities involving the MLL gene. This led us to speculate that maternal exposure during pregnancy to environmental agents that inhibit DNA topo 2 may be associated with the development of leukemia in infants. DNA topo 2 inhibitors have been found in specific fruits and vegetables, and in soy, coffee, wine, tea and cocoa, as well as in certain pesticides, solvents and medications. In a preliminary study, we reinterviewed mothers of infant cases and their matched controls who had participated previously in 1 of 3 epidemiologic studies of childhood leukemia conducted by the Children's Cancer Group over a 10-year period. We evaluated potential DNA topo 2 inhibitor exposure through maternal diet and medications. Of the 84 original matched sets who were reinterviewed, there was no positive association with increasing maternal consumption of DNA topo 2 inhibitor containing foods either for the overall group or for infants in the acute lymphoblastic leukemia stratum. However, there was an approximately 10-fold higher risk of infant AML with increasing maternal consumption of DNA topo 2 inhibitor-containing foods. The assay to screen environmental agents that inhibit DNA topo 2 has been established and new inhibitors are being identified routinely. PMID- 9876474 TI - Review of the evidence for an association between infant feeding and childhood cancer. AB - To assess the association between infant feeding and childhood cancer, a qualitative review of 9 published case-control studies was undertaken. The results of this synthesis suggest that children who are never breast-fed or are breast-fed short-term have a higher risk than those breast-fed for > or = 6 months of developing Hodgkin's disease (HD), but not non-Hodgkin's lymphoma or acute lymphoblastic leukemia. HD has features of a complex cellular immune disorder and of chronic infection. Human milk contains an extensive array of anti microbial activity and appears to stimulate early development of the infant immune system. Artificially fed infants negotiate exposure to infectious agents without the benefits of this immunologic armament and do not do as well as breast fed infants in resisting infection. Thus, human milk may make the breast-fed infant better able to negotiate future carcinogenic insults by modulating the interaction between infectious agents and the developing infant immune system or by directly affecting the long-term development of the infant immune system. Further research should attempt to confirm the association between infant feeding and HD in large, population-based, case-control studies. Improved measurement of infant feeding must be addressed if future studies are to advance our understanding of this association. In addition, studies of specific measures of immunity, particularly of cellular immune responses, should be conducted in populations of breast-fed and non-breast-fed young children. PMID- 9876475 TI - Bone and mineral abnormalities in childhood acute lymphoblastic leukemia: influence of disease, drugs and nutrition. AB - In children with acute lymphoblastic leukemia (ALL), abnormalities in mineral homeostasis and bone mass were first reported by our group in the late 1980s. Prospective longitudinal cohort studies in 40 consecutive patients receiving treatment according to the Dana-Farber Cancer Institute (DFCI) protocol 87-001 and 16 children receiving DFCI protocol 91-001 afforded us the opportunity to explore various etiologies of the observed abnormalities in mineral and bone metabolism, specifically the leukemic disease process and chemotherapeutic drugs such as steroids and aminoglycoside antibiotics. At diagnosis of ALL, > 70% of children had abnormally low plasma 1,25-dihydroxyvitamin D, 73% had low osteocalcin and 64% had hypercalciuria, indicating an effect of the leukemic process on vitamin D metabolism and bone turnover. During remission induction, treatment with high-dose steroid (prednisone or dexamethasone) resulted in further reduction in plasma osteocalcin and elevated parathyroid hormone levels. During 24 months of chemotherapy-maintained remission, reduction in bone mineral content (BMC), as measured by Z-scores, occurred in 64% of children, most severely affecting those > 11 years of age. A reduction in BMC during the first 6 months had a positive predictive value of 64% for subsequent fracture. By the end of 2 years of therapy, fractures occurred in 39% of children and radiographic evidence of osteopenia was found in 83% of the entire study group. Investigations of the biochemical basis of the bone abnormalities revealed that by 6 months hypomagnesemia developed in 84% of children (of whom 52% were hypermagnesuric) and plasma 1,25-dihydroxyvitamin D remained abnormally low in 70%. Altered magnesium status was attributed to renal wastage of magnesium following cyclical prednisone therapy and treatment with aminoglycoside antibiotics such as amikacin for fever accompanying neutropenia. Dietary intake and absorption of magnesium were normal. In 10 children treated for hypomagnesemia with supplemental magnesium for up to 16-20 weeks, plasma magnesium normalized in only 50% of subjects. PMID- 9876476 TI - Bone mass and body composition after cessation of therapy for childhood cancer. AB - Our aim was to review current information on body composition and bone mass after cessation of therapy for childhood cancer and to present preliminary data on body composition and bone mass in a group of Danish survivors of childhood leukaemia or lymphoma. Elevated body-mass index (weight/height2; BMI) is frequent after treatment for childhood acute lymphoblastic leukaemia. BMI increases during therapy or within the first year after therapy and remains abnormal thereafter. Treatment with corticosteroids, abnormal growth-hormone secretion after treatment with cranial irradiation (CI) or corticosteroids, younger age at diagnosis, or female gender were risk factors for elevated BMI in earlier studies. We evaluated 185 survivors of childhood leukaemia or lymphoma by dual-energy X-ray absorptiometry scanning. We found elevated whole-body relative fat mass, which was associated with CI. Other studies found reduced bone mass in the radius, the lumbar spine and the whole body after treatment for childhood cancer. Growth hormone deficiency that is not adequately corrected, CI, reduced height or reduced weight were risk factors for reduced bone mass. In our 185 participants, the whole-body bone mass was also reduced significantly compared with reference values. CI and older age at follow-up were risk factors for reduced bone mass. We conclude that the elevated relative fat mass and reduced bone mass seen after treatment for childhood leukaemia or lymphoma is associated mainly with CI. PMID- 9876477 TI - Bone mineral density in long-term survivors of childhood cancer. AB - Bone mineral density (BMD) of the lumbar spine was measured in 97 long-term survivors of childhood cancer 5-23 years after diagnosis using dual-energy X-ray absorptiometry (DXA). They had been treated for acute leukemia (n = 22), brain tumors (n = 16), lymphomas (n = 16), Wilms' tumor (n = 10), neuroblastoma (n = 7) and other cancers (n = 26). The correlations between BMD and the Z-scores for weight for height, height for age and weight for age at diagnosis and follow-up were evaluated with stepwise multiple regression. Correlations with cumulative corticosteroid and radiation dose were examined with Spearman's correlation coefficient. The number of nature of fractures were noted. A BMD Z-score of below -2 was present in 13 and a BMD Z-score of -1 to -2 in 31 children. In total, a low BMD was observed in 45% of children. Height for age at follow-up correlated significantly with BMD Z-score. Increasing doses of cranial irradiation (18-54 Gy) were associated with lower BMD (p = 0.001, Spearman). This was true also for 22 children with acute lymphoblastic leukemia (ALL) who had received 18-24 Gy cranial irradiation (p = 0.04, Spearman). Fractures occurred in 14 children following trauma. The difference in BMD Z-scores of children with and without fractures did not achieve statistical significance although the majority of the children with fractures had low BMD Z-scores. The significant inverse correlation between height for age at follow-up and BMD must be interpreted with the realization that DXA is not a volumetric measurement of BMD and that short stature is associated with a smaller skeletal mass. PMID- 9876478 TI - Impact of nutrition on pharmacokinetics of anti-neoplastic agents. AB - It has been estimated that approximately 80% of the world's pediatric population lives in countries with limited resources, and that 43% of these children are malnourished. In children with cancer, malnutrition may antedate the diagnosis or be a result of aggressive chemotherapeutic regimens. Studies have shown that children with cancer and malnutrition have a less favorable prognosis, a higher risk of early relapse, and tolerate chemotherapy poorly when compared with children with normal nutritional status. Improvements in nutritional status may improve tolerance to chemotherapy. An understanding of the mechanisms responsible for the effects of malnutrition on drug disposition and pharmacodynamic response is important, especially for anti-neoplastic agents, which have a narrow therapeutic index and may be associated with potentially severe or life threatening side-effects. Several factors related to malnutrition have been suggested to alter drug disposition. Diminished protein "status" in malnourished children results in lower amounts of plasma proteins, increasing the concentration of free drug available to exert its cytotoxic effect. Severely malnourished individuals also exhibit decreased oxidative metabolism and reduced glomerular filtration rate (GFR), potentially increasing concentrations of parent drug or active metabolites. Malnourished children receiving chemotherapy for the treatment of an underlying malignancy may need specifically "tailored" protocols to achieve therapeutic response while minimizing adverse acute and long-term side effects. The role of specific interventions, such as correction of nutritional status or pharmacokinetic drug monitoring, should be evaluated in this context. PMID- 9876479 TI - Nutritional status and socio-economic conditions as prognostic factors in the outcome of therapy in childhood acute lymphoblastic leukemia. AB - The majority of children on earth are to be found in the developing world, many of them malnourished members of impoverished families. Thus, the effects of socio economic status (SES) on the therapeutic response of children with cancer are obviously relevant. The outcome of treatment in patients with the commonest form of cancer in childhood (acute lymphoblastic leukemia, ALL) is clearly related to their SES. Studies conducted mainly in developing countries have shown malnutrition to be an important prognostic factor in such children. However, other socio-economic conditions could affect the outcome of therapy in patients with ALL: access to communications, transportation, laboratory studies and therapy. Even in children with an "adequate" SES, malnutrition is still an adverse prognostic factor. Nutritional supplementation appears to be a valuable addition to chemotherapy in undernourished children with ALL. The choice of treatment for these children should accommodate the cultural, economic and nutritional status of the patients and their families. Protocols must be created for testing methods of nutritional intervention and their influence on pharmacology, drug tolerance and survival in ALL. The influences of poverty and illiteracy on compliance with treatment, especially oral medication, need to be evaluated. Such investigations are essential to improve results of treatment of socio-economically disadvantaged children suffering from ALL and other forms of cancer. PMID- 9876480 TI - Low socioeconomic status is a strong independent predictor of relapse in childhood acute lymphoblastic leukemia. AB - The results of the treatment of acute lymphoblastic leukemia (ALL) in children depend not only on the biologic diversity of the leukemia cell, the multi-drug treatment schedule and the individual variability of drug metabolism, but also on the socioeconomic and cultural background of the leukemic child. Social and cultural disparity is very marked in underdeveloped countries and has been increasing in industrialized nations. The prognostic influences of these factors are poorly documented and sometimes mistakenly attributed to differences in ethnic origin. We have investigated in Brazil the relative impact of malnutrition and socioeconomic status on the outcome of ALL, adjusting for the known influence of biologic factors. Children with ALL (n = 167) treated with a Berlin-Frankfurt Munster-based protocol were studied prospectively. At a median follow-up of 1623 days, the estimated probability of disease-free survival was 43 +/- 4%. The main cause for interruption of remission was bone-marrow relapse. Socioeconomic indicators of poverty (poor housing conditions, low per capita income and energy consumption) were significantly associated with a greater risk of relapse in univariate analysis. They were consolidated in a single index, socioeconomic status (SES), defined by the product of monthly per capita income times mean familial daily energy consumption. Other unfavorable findings included age, z score for the height for age at diagnosis (HAZ) below-1.28 and the z score for weight for age below-1.28. After adjustment in Cox's multivariate model, only HAZ and poor SES remained as predictive factors for relapse. Poor prognosis for leukemic children of low SES is just another indicator of social inequality. PMID- 9876481 TI - Aspects of altered metabolism in children with cancer. AB - Severe weight loss associated with cancer continues to be a major cause of morbidity in cases of childhood malignancy. The etiology is not completely understood but is probably multifactorial, including reduced ingestion and altered metabolism of nutrients. Changes in the host metabolism of protein, fat and carbohydrate in the cancer-bearing host have been demonstrated both in animal models and in patients. Changes include increased protein turnover and loss of the normal compensatory mechanisms seen in starvation. Additionally, increased lipid breakdown results in depletion of lipid stores and changes in carbohydrate metabolism result in an energy-losing cycle. The increase in protein turnover seen in children with leukemia may be related to the tumor, the chemotherapy administered or to related conditions such as febrile neutropenia. The role of endogenous mediators of cancer cachexia has not yet been clearly elucidated, although tumor necrosis factor, interleukin I and interleukin 6 appear to be involved. Studies of energy expenditure in children with cancer have indicated that certain patients with a raised metabolic rate are at particular risk of severe weight loss. The challenge is to identify these vulnerable patients and to provide adequate nutritional support early in treatment and therefore avoid the deleterious effects of cachexia. PMID- 9876482 TI - Benefits of nutritional intervention on nutritional status, quality of life and survival. AB - Most cancers in children are acute diseases. Therefore, the incidence of malnutrition, in general, is not different from the incidence in the referral population. Some specific tumors, such as neuroblastoma and those resulting in the diencephalic syndrome, can be exceptions. By contrast, malnutrition is a frequent problem during modern intensive cancer treatment as the result of the associated anorexia, altered taste sensations and catabolic effects of drugs. In addition, there are psychogenic factors and metabolic consequences associated with the tumor itself. Nutritional support does improve the feeling of well-being and performance status, while maintaining or improving the immune competence, thereby potentially affecting survival by limiting infectious episodes. There is no convincing evidence to date that nutritional support has an antineoplastic effect per se, but deficiency of a specific nutrient might be beneficial because of a differential requirement between tumor and normal cells. Theoretically, nutritional support might enhance tumor growth but also susceptibility to chemotherapy. In either case, nutrition is a support modality that must be given with appropriate tumor-directed therapy if curative intent is the goal of treatment. Nutrition remains a consideration after therapy is completed. This generates different challenges. If further tumor-directed therapy is futile, the decision to continue nutritional support is difficult, but if the child is well, nutritional rehabilitation must be pursued. Finally, the cured child continues to benefit from dietary advice. Nutrition should be viewed for what it is: supplying the most basic need of children. PMID- 9876483 TI - Alternative nutritional cancer therapies. AB - Increasing attention is being paid to the role of nutrition in cancer. Dietary measures, such as decreased consumption of calories, fat, alcohol and smoked or pickled foods have been shown to reduce the incidence of specific "adult" cancers, while increased dietary fiber appears to have a protective role. However, no clear scientific evidence exists that dietary manipulation is a successful primary therapy for established cancer. A significant percentage of adult and child cancer patients take unproven therapies during their illness. Alternative nutritional therapies, of which there is a wide variety, are the commonest of these reflecting current public interest in "natural" remedies. The efficacy and potential toxicity of commonly utilized dietary therapies are here reviewed, in particular the macrobiotic philosophy, the Gerson diet, the Livingstone diet, and the use of vitamin and mineral therapy. While details may differ, most alternative approaches involve fresh whole foods, with strong emphasis on low-fat vegetarian diet. Most are nutritionally adequate, at least for adults. No anti-cancer diet has been shown to cure established cancers, even those whose incidence is decreased by dietary changes. Careful dietary manipulation may at least improve quality of life for adult cancer patients, and, together with conventional therapy, may prolong survival in selected cancer patients. Assessment by carefully controlled prospective clinical trials is essential; those in pediatric patients must be controlled very strictly, since tumors in children have not been shown to be influenced by diet, and the diets described may be inadequate for children with malignant disease. PMID- 9876484 TI - Aggressive oral, enteral or parenteral nutrition: prescriptive decisions in children with cancer. AB - Over the past 18 years, our laboratory has been interested in the pathogenesis of energy imbalance caused by a variety of diseases. Our view is that a clear understanding of the various factors causing negative energy balance, which in turn results in malnutrition, is the most effective way of designing preventive and therapeutic nutritional strategies. Thus, in cancer, one of the common factors is anorexia, due either to the primary tumor or to the effects of cancer therapy. Currently there is little evidence of increased resting energy expenditure in children with cancer, except in cases with very high tumor burden. Conversely, there are suggestions of a failure to down-regulate resting energy expenditure in the presence of reduced food intake in patients with cancer. Damage to the gastrointestinal tract, due to the effects either of the tumor or of tumor therapy, may result in maldigestion and/or malabsorption. Thus, as a result of a combination of reduced intake, reduced absorption and increased needs, the child with cancer may become malnourished. Prevention and treatment are dependent on the type of cancer and the pathogenesis of the negative energy balance. In broad terms, we try as far as possible to use external routes. With the advent of percutaneously placed gastrostomies and gastrojejunal tubes, we use these methods increasingly to provide nutritional support. Only in patients whose gastrointestinal tract cannot be used do we turn to i.v. feeding. In these patients, the placement of a central venous line is required, but great care must be taken to avoid infection. Whatever form of nutritional support is used, whether enteral or parenteral, we measure the body composition and energy expenditure in the patient, so that the nutritional therapy can be tailored to the child's specific needs. Using these approaches, we are having significant success in preventing and reversing malnutrition in children with cancer and those undergoing bone-marrow transplantation. PMID- 9876485 TI - Algorithm for nutritional support: experience of the Metabolic and Infusion Support Service of St. Jude Children's Research Hospital. AB - The Metabolic and Infusion Support Service (MISS) at St. Jude Children's Research Hospital was established in 1988 to improve the quality of nutritional support given to children undergoing therapy for cancer. This multidisciplinary group, representing each of the clinical services within the hospital, provides a range of services to all patients requiring full enteral or parenteral nutritional support. In 1991, the MISS developed an algorithm for nutritional support which emphasized a demand for a compelling rationale for choosing parenteral over enteral support in patients with functional gastrointestinal tracts. Compliance with the algorithm was monitored annually for 3 years, with full compliance defined as meeting all criteria for initiating support and selection of an appropriate type of support. Compliance rates were 93% in 1992, 95% in 1993 and 100% in 1994. The algorithm was revised in 1994 to include criteria for offering oral supplementation to patients whose body weight was at least 90% of their ideal weight and whose protein stores were considered adequate. Full support was begun if no weight gain occurred. Patients likely to tolerate and absorb food from the gastrointestinal tract were classified into groups defined by the absence of intractable vomiting, severe diarrhea, graft-vs.-host disease affecting the gut, radiation enteritis, strictures, ileus, mucositis and treatment with allogeneic bone marrow transplant. Overall, the adoption of the algorithm has increased the frequency of enteral nutritional support, particularly via gastrostomies, by at least 3-fold. Our current emphasis is to define the time points in therapy at which nutritional intervention is most warranted. PMID- 9876486 TI - Growth and body composition in response to chemotherapy in children with acute lymphoblastic leukemia. AB - Severely malnourished children afflicted by acute lymphoblastic leukemia (ALL), particularly in developing countries, have reduced tolerance to chemotherapy and a compromised prospect for survival. We investigated the prevalence and severity of alterations in growth and nutritional status in children with ALL from population-based referral areas in Canada. All children were treated with Dana Farber Cancer Institute ALL Consortium protocols. First, the relative impact of cranial irradiation (CI) and chemotherapy on growth was studied in 116 children at diagnosis and at 6-month intervals during treatment. We observed a decline in height standard deviation (SD) score in the first year in all children, and a further decline in height SD score during the second year only in the children who received CI. Weight reduction occurred in the first year, but during the second year there was a disproportionate increase in weight compared with height, suggesting that children treated with ALL have a tendency toward obesity. Both chemotherapy and CI contribute to the altered growth observed in children treated for ALL. Second, intestinal functional integrity was assessed in 16 children during post-induction chemotherapy. Nutrient intake was adequate and there was minimal evidence of malabsorption: fat malabsorption occurred in only 1 child (after treatment-related pancreatitis), abnormal D-xylose absorption occurred in 2 children at 6 months of therapy (returning to normal 6 months later) and abnormal lactose absorption occurred in 4 children. Third, weight, height, whole body lean and fat mass measured by dual-energy X-ray absorptiometry and serum albumin were determined at diagnosis and at 6-month intervals throughout therapy in 19 children with ALL. Height SD scores decreased significantly during treatment. Serum albumin was abnormally low in 6/19 at diagnosis and 14/18 during intensive consolidation therapy. The mean change in the ratio of lean mass to total body weight showed a 5% reduction by 6 months of therapy. Body fat increased from a mean of 22% at diagnosis to 28% at completion of therapy. The majority of children treated for ALL thus have significant changes in nutritional status manifested by reductions in growth, alterations in lean and fat body mass and abnormally low serum proteins during intensive therapy. PMID- 9876487 TI - Diet that prevents cancer: recommendations from the American Institute for Cancer Research. AB - The current epidemiological transition in less developed countries is resulting in an epidemic of chronic diseases, with cancer being the second most common cause of mortality. The evidence linking diet to the development of cancer is based largely on epidemiological analysis of the relationships of the frequency of different cancers to data on food consumption. Cohort results have made clearer the link between diet and cancer, as have data on a number of biological mechanisms. Based on the available data, recommendations on dietary practices that may prevent cancer have been published recently by the American Institute for Cancer Research and the World Cancer Research Fund. Key recommendations are: diet should be based on plant products; 400 g of vegetables and fruits, to provide more than 10% of energy, should be consumed daily; cereals, legumes and tubers should provide at least 50% of energy, and sugars less than 10%; no more than 80 g of meat should be consumed, preferably fish or poultry, and limited amounts that are cured or smoked; fat intake should be limited to no more than 30% of energy, with a predominance of monounsaturated and polyunsaturated forms; total salt consumption should be less than 6 g; perishable foods should be kept frozen or refrigerated and consumed promptly; foods should be cooked at low temperatures, better to be boiled or steamed rather than fried or grilled; alcohol should not exceed 2 drinks a day. In addition to these dietary guidelines, cancer prevention may be achieved by not smoking, by avoiding excess weight, and by increasing physical activity, including half an hour of exercise and 4 hr not resting in a chair or bed. PMID- 9876488 TI - [Effect of x-irradiation on the properties of cyclic AMP-dependent protein kinases from rat spleen lymphocytes]. AB - It was shown, that in conditions of acute radiation affection in doze 0.5 and 1 Gy, there was a decrease of 32 and 55% in activity of cyclic AMP-dependent protein kinases, isolated from cytosol of lymphocytes. The analysis of cAMP dependent protein kinases properties has shown, that disturbance in the interaction of the enzyme with protein substrate of phosphotransferase reaction and main modulater enzymes activity--cAMP proceeds of the ionising radiation effect. PMID- 9876489 TI - [Effect of tonicity of the medium on the sensitivity of Escherichia coli bacteria to gamma-quantum 60Co, ultrasound and hyperthermia]. AB - The cell lethality and permeability induced in Escherichia coli B/r and Escherichia coli BS-1 by 60Co gamma-ray irradiation, ultrasound and hyperthermia in media containing different concentrations of NaCl have been investigated. It was shown that independently from the nature of damaging factors hypotonic media increase while hypertonic media in certain range of osmolyte concentration decrease sensitivity of cells to action of this factors. It was proposed that discovered phenomenology was caused by salt modification of status of the cell osmotic homeostasis destabilizing by ionizing radiation, ultrasound or hyperthermia and was not related with the system of dark repair of DNA. PMID- 9876490 TI - [Characterization of the adaptive response to the action of gamma-rays, induced by low doses of 14C in Chinese hamster fibroblasts]. AB - It has been studied the correlation of the mitotic activity of the chromosome aberrations and apoptosis, in the V-79 cells pre-exposure to an adapting dose of ionizing radiation from 14C-thymidine prior to an acute challenge dose of gamma rays. In spite of that the incubation of the cells with isotope increased of the yield of the chromosome aberrations, but the cells became more resistant to following gamma-irradiation. Increasing the adaptive dose of the 14C on degree didn't influence on the present of the adaptive response. However, using concentrations of the 14C damaged metaphase/anaphase transition and cells blocked in this check-point by apoptotic death. The results suggest, that the cellular selection has been involved in 14C-induced adaptive response, estimated by level of asymmetric chromosome aberrations in V-79 cells. PMID- 9876491 TI - [Features of radiation carcinogenesis in man at low doses and low dose rates]. AB - The accepted linear no-threshold (LNT) dose response for stochastic effects is not valid even for the cellular level. Published data on the human radiation carcinogenesis demonstrate now the radiation hormesis or the absence of effects or its reduction in the whole low dose interval when a dose rate is decreased. This is demonstrated for leukemia and for lung, breast, thyroid, bone, skin and liver solid cancers, for such organs, which are responsible in the more, than 1/2 of cancer detriment postulated by the ICRP. It is possible valid for the whole of solid cancers and for nontumorogenic death. PMID- 9876492 TI - [Protective action of leukinferin during radiotherapy of oncologic patients]. AB - In clinical observation on 248 patients with breast cancer IIb-III-b stages given radiotherapy on breast gland, parasternal and axillary-subclavian regions (total doses 40 Gr for each) before mastectomy was shown, that use LF im 2-3 times a week through the course of therapy decreases statistically significant pancytopenia, induced by radiation. Quantity leukocytes was even increase in a process of therapy. Frequency of anemia and lyphopenia was decreased almost 3 times. E-RFC were depressed also more slight, then in the group without immunocorrection and after mastectomy practically, restored. IFN-gamma production by T-lymphocytes was not changed and essentially more high then in the comparative group. The presented data are shown, that LF provides a defensive action on hematological indices and immune effectors function in oncological patients during radiotherapy. PMID- 9876493 TI - [Production of tumor necrosis factor (TNF) by macrophages and T-lymphocytes of rodents as affected by acute gamma-irradiation]. AB - TNF production in peritoneal macrophages and in splenic T cell from mice and ground-squirrels were performed after acute in vivo and in vitro doses of ionizing radiation ranging from 0.1-6 Gy. It was observed that doses in the range 0.5-6 Gy induced the increase of TNF production along with the decrease of mitogen-induced proliferation in T lymphocytes. PMID- 9876494 TI - [Separate and combined effects of ionizing radiation and cyclophosphamide on thymus-dependent humoral immune response]. AB - The total exposure of gamma-irradiation, (1-2 Gy) or intraperitoneally injection of cyclophosphamide (10-50 mg/kg) resulted in the dose-dependent inhibition of humoral immune response. The immunosuppression was expressed to a greater degree in lymphatic nodules, than in spleen. The combined action of these factors showed the effect of synergism. The injection of cyclophosphamide in mice neutralized the effect of antibody genesis induced by low dose irradiation (0.1 Gy). PMID- 9876495 TI - [Dynamics of the changes in various integral indices of the functional state of the body of white rats during the prolonged action of x-radiation and external heat]. AB - Effects of low doses of X-radiation (0.02 Gr. a session, for 25 days), external heat (daily 4-h exposure in a thermochamber at 37.0 degrees C) and combined (consecutive) action of these factors on mature Wistar male rats were studied. Experimental animals were somewhat behind in body mass increment during exposure to the physical factors as compared to control rats. Radiation promoted a rise in rectal temperature, external heating to its small decrease, while combined exposure to the factors levelled the effects. There were certain changes in the CNS, as indicated by a small increase in the summation-threshold index. The heart rate did not significantly change. The changes normalized within 1-5 weeks after cessation of the exposures. PMID- 9876496 TI - [Analysis of the strategy for the use of protective measures in agriculture after the accident at the Chernobyl power plant]. AB - Justification of the methodological approach to evaluating effectiveness of the countermeasure strategies in agricultural production on contaminated territories is provided. The results from a comparative analysis of strategies for protective measures in agriculture after the Chernobyl NPP accident are presented. The time between radioactive fallout and application of countermeasures is shown to be one of the main factors responsible for effectiveness of countermeasures applied. PMID- 9876497 TI - [Biological effects in natural populations of small rodents in radiation-polluted territories. Description of sites. Dynamics of concentration of gamma-emitting radionuclides in populations of two species of small mammals]. AB - It has been revealed that dynamics of gamma-emitting radionuclide concentration in consecutive generations of two species of wild rodents (European bank vole and yellow-necked mouse) is characterised by the phases of an increase, a maximum content (peaks) and a decrease over 10 years after the Chernobyl accident. The peaks of specific activity of gamma-emitting radionuclides in populations in the areas with different densities of radio-contamination falls not on the first year but on the next ones (1987-1989) after the catastrophe, i.e. are observed in the subsequent (3-8) generations of animals. The revealed shift of maximum of radionuclide concentration in comparison with the maximum of their fallout is likely caused by an increase in radionuclide biological availability. PMID- 9876498 TI - [Biological effects in natural populations of small rodents in radiation-polluted territories. Dynamics of chromosome aberration frequency in a number of generations of European bank vole (Clethrionomys glareolus, Schreber)]. AB - The dynamics of chromosome aberration frequency in bone marrow cells of many generations (14) of bank vole living in the radioactive trace of the Chernobyl catastrophe (1986-1992) has been analysed. The study revealed that the chromosome aberration frequency in voles in the areas with radio-contamination density 220 and 1526 kBq/m2 (for 137Cs) significantly exceeds the control level 3-7 times over the whole period under investigation. The dynamics of the frequency of structural chromosome injuries from 1986 to 1991-1992 is characterised by the tendency to increase in all populations inhabiting the areas with various radio contamination density (8-1526 kBq/m2). PMID- 9876499 TI - [The ecological cause of the higher accumulation of 90Sr and 137Cs in the food of northern inhabitants]. AB - The higher accumulation of 90Sr and 137Cs in the trophic chains of North inhabitants was analysed. The specific of the higher accumulation is caused by one-sided trend of the typical for North the biotical and abiotical factors. PMID- 9876500 TI - [Analysis of pulsed bioelectric activity of rabbit cerebral cortex in response to low-intensity microwave radiation]. AB - In experiments on 22 rabbits the influence of a pulse microwave irradiation on extracellular activity of separate nervous cells of sensorimotori and occipital areas of a cortex brain is shown. The reaction could consist in activation or in braking frequency of the discharges, that was connected to frequency impulsation in an initial background. The researched mode of a microwave irradiation (1.5 GHz, duration of a pulsed-0.4 microsecond, frequency of their recurrence 1000 Hz, DFEpulsed-300 microW/sm2) had a corrigizing action. PMID- 9876501 TI - [Changes in brain memory functions as affected by a constant magnetic field]. AB - Effects of CMF on memory processes in adult volunteers (non-sleep us hypnotic condition) has been studied at laboratory and clinical conditions. The results point out that CMF changes the interhemispheric relations and result in memory malfunction. The malfunction of memory activity in volunteers after action of CMF was influenced by two factors: magnetic field position under different brain areas and functional status of volunteers: awake or hypnotic. PMID- 9876502 TI - Recent approaches in insulin delivery. AB - Insulin remains indispensable in the management of diabetes mellitus since its discovery in 1921. The foreignness of early available porcine and bovine insulin led to the development of human insulin by transpeptidation and biosynthesis in microorganisms. Needle phobia and stress of multiple daily injections led to the investigation and exploitation of all promising routes, ranging from nasal to rectal, by a wide variety of devices and delivery systems. This article describes the development of human insulin, various routes for delivery of insulin (including oral, nasal, buccal, rectal, and pulmonary), and various devices for regulated, safe, and convenient insulin delivery. The article reviews some recent advances in insulin delivery such as the bioresponsive and self-regulated insulin delivery system. PMID- 9876503 TI - Evaluation of dosage forms. IV. Studies on commercial phenylbutazone tablet dosage forms. AB - In order to determine the feasibility of dissolution-dialysis as a suitable technique for in vitro evaluation, studies on commercial phenylbutazone tablets were carried out. Although disintegration time and dissolution parameters did not give a true indication of bioavailability, an excellent correlation was obtained between the dialysis rate constant (K) and various pharmacokinetic parameters obtained from bioavailability studies on human volunteers. PMID- 9876504 TI - Rheology and filling characteristics of particulate dispersions in polymer melt formulations for liquid fill hard gelatin capsules. AB - The rheology and capsule filling properties of molten excipients, Dynafill, Dynasan-114. Lutrol-F68, and polyethylene glycols (PEG) 6000, 8000, 10,000, and 20,000 have been investigated. Lactose (alpha-monohydrate) was selected as a model particulate solid with low solubility in PEG in order to investigate the effects of disperse phase particle size, concentration, and PEG molecular weight on rheology and capsule filling properties of these systems. All excipients behaved as Newtonian fluids between 65 and 90 degrees C, which was chosen as a possible temperature range for liquid filling of hard gelatin capsules. The excipients, apart from Dynasan-114 and PEG 20,000, showed satisfactory capsule filling properties at 70 degrees C using a semi-automatic filling machine. Dynasan-114 (viscosity = 0.012 Pa.s at 70 degrees C) leaked from the seals between the hopper and pump of the filling machine, whereas PEG 20,000 (viscosity = 24 Pa.s at 70 degrees C) showed bridging of the molten polymer between successive capsule bodies during the filling process. The effect of disperse phase (lactose) particle size and concentration, and continuous phase (PEG) molecular weight on the apparent viscosity and filling properties of the non Newtonian dispersions were investigated at 70 degrees C. Satisfactory filling of the dispersions was achieved at 70 degrees C up to a limiting concentration of disperse phase which was dependent upon disperse phase particle size and continuous phase molecular weight, and corresponded to a pronounced increase in apparent viscosity of the dispersion. PMID- 9876505 TI - Effect of compression speed and pressure on the physical characteristics of maltodextrin tablets. AB - The present paper studies the effect of applied pressure (0-300 MPa) and compression speed (8 and 40 cycles/min) on the physical characteristics of four varieties of maltodextrins for direct compression. The materials were tableted by using a single-punch tablet machine. On the basis of the mechanical properties it seems to be reasonable to propose a limit in the plastic deformation and consequently bonding between particles. This limit could be approximately 90 MPa, and it was almost independent of the variety compressed. Disintegration behavior and, most probably, release properties are related with this limit; i.e., mechanical parameters and disintegration time increased as applied pressure was increased up to this limit. Above this limit, no differences of note were found. The different movement of particle layers in the single-sided eccentric profile led to the computation of differences between upper and lower tablet hardness surfaces, which were indicative of a consolidation mechanism. The differences obtained were indicative of the plastic deforming nature of maltodextrins. PMID- 9876506 TI - Evaluation of a novel phase separation technique for the encapsulation of water soluble drugs in biodegradable polymer. AB - Biodegradable microcapsules of four water-soluble drugs (pentamidine, captopril, diltiazem, and metoprolol) were prepared using a novel phase separation technique. The microcapsules prepared by this method were irregular in shape. Particle size of the microcapsules was between 60 and 500 microns. The efficiency of encapsulation for all four drugs was more than 40% at 20% drug loading. The encapsulation increased up to 75% or higher when the drug loading was reduced to 5%. The in vitro dissolution was drug dependent. The microcapsules prepared with 5% drug loading showed the minimum dissolution at the end of 24 hr. The initial drug release increased significantly when the drug loading was increased up to 20%. Although the formulations containing 5% drug maintained a sustained-release profile up to 45 days, the formulations containing 10 and 20% drug released more than 50% drug within the first 2 days. PMID- 9876507 TI - Effect of various lipid-bile salt mixed micelles on the intestinal absorption of amphotericin-B in rat. AB - Amphotericin-B (AmB), a lipophilic polyene antibiotic, is the drug of choice in the treatment of many serious mycotic diseases. The solubility and gastrointestinal membrane permeability (Papp) of AmB in mixed micellar systems were examined. Membrane permeability was determined using a rat gut perfusion method. The mixed micellar systems studied contained the bile salt in association with fatty acid. All mixed micellar systems enhanced the absorption of AmB relative to the simple micelle. These results have shown that mixed micelles can enhance the absorption of AmB to a greater extent relative to nonmicellar and simple micellar systems. Maximum enhancement (> 20-fold) in the rate of AmB absorption was obtained with the sodium desoxycholate/soya lecithin 40:40 mM system. These results offer a possible explanation for the reported enhancement in gastrointestinal absorption of AmB when coadministered with lipid-bile salt mixed micelles, and these systems can be used as a vehicle for designing novel drug delivery systems for poorly absorbable drug(s). PMID- 9876508 TI - Particle size distributions from multiparticulate dissolution. AB - It has been demonstrated theoretically that the particle size distribution of particles in a given sieve fraction of a powder may be assessed by means of short term dissolution data. Theoretical considerations in this article show that by accounting for polydispersity in a powder sample, the cubic expression in time for amount undissolved and fraction undissolved gives rise to integrals that are essentially moments of the distribution function of one of the defining dimensions of the particle. The first and the second moments can be used to calculate the distribution parameters, (mean and standard deviation) of such a dimension of a crystalline powder. The theory is based on a model geometry, a parallelepiped, for the description of particles such as needles, plates, and prisms. The theory is substantiated by experimental data. A method for obtaining the particle size distribution parameters from the results of dissolution of three sieve fractions of oxalic acid dihydrate, and the general application of this to particle size determination is discussed. To validate the method, the distribution of lengths and breadths of oxalic acid dihydrate particles was obtained from microscopy. From actual powder dissolution data, an estimate of the mean height-to-breadth ratio of these particles belonging to a certain sieve fraction was obtained. With the knowledge of the dissolution rate constant, K, for oxalic acid dihydrate under specified hydrodynamic conditions, it was possible to evaluate the moments of the distribution function. The distribution parameters so obtained were in good agreement with the results obtained from microscopy. PMID- 9876509 TI - Effects of drug solubility, drug loading, and polymer molecular weight on drug release from Polyox tablets. AB - This study investigated the effects of polymer molecular weight, drug solubility, addition of a water-soluble excipient, and drug loading on zero-order release kinetics and elucidated the release mechanism of a drug from directly compressed tablets. Directly compressed tablets consisting of polyethylene oxides (PEO) (MW = 0.9, 2.0 and 4.0 x 10(6)) and drugs (solubility ranging from 290 to 25,000 mg/l) were formulated with or without a water-soluble excipient (lactose). For PEO tablets (MW = 0.9 x 10(6)), drug release is primarily swelling/erosion controlled for drugs for which solubility is below 1%, resulting in zero-order release kinetics. For PEO tablets (MW = 4.0 x 10(6)), drug release is controlled at a zero-order rate by the dissolution rate of the drug at high loading (39%). At low loading (20%), drug diffusion through the swollen gel layer becomes the governing release mechanism. For a highly water-soluble drug (e.g., diclofenac Na), drug diffusion is the controlling mechanism regardless of the molecular weight of the PEOs. Zero-order release kinetics can be achieved with PEO tablets (MW = 0.9 x 10(6)) for drugs for which solubility is below 1%. PEO tablets (MW = 2.0 x 10(6)) provided zero-order release for poorly water-soluble drugs (below 0.2%) at 39% drug loading. It is possible to attain zero-order release kinetics with PEO tablets (MW = 4.0 x 10(6)) using a drug which has a solubility of less than 0.1%. PMID- 9876510 TI - Interactions between lonidamine and beta- or hydroxypropyl-beta-cyclodextrin. AB - The possibility of obtaining inclusion complexes between lonidamine and beta- or hydroxypropyl-beta-cyclodextrin have been evaluated by phase solubility diagram, differential scanning calorimetry (DSC), and x-ray diffractometry. The applied complexation methods were spray-drying, kneading, and solid dispersion. DSC and x ray analyses of the powders revealed an external interaction between lonidamine and cyclodextrins. Dissolution profiles of the obtained powders were also studied to define the most appropriate preparation method and molar ratio to use in attempts to increase lonidamine water solubility. PMID- 9876511 TI - Formulation and in vitro and in vivo availability of diclofenac sodium enteric coated beads. AB - Diclofenac sodium enteric-coated beads were prepared using the conventional pan coating technique. Eudragit L100 was used as a pH-dependent release-controlling polymer. The beads were evaluated for their particle size distribution, drug loading efficiency, flowability, in vitro release in 0.1 N HCl (pH 1.2) and phosphate buffer (pH 6.8), and bioavailability in beagle dogs relative to the commercial enteric-coated tablets Voltaren. The beads showed a narrow particle size distribution in which 83% of the beads were in the range of 1-2 mm. The actual yield of the beads was 90.5% and their drug loading was 92%. The beads released about 8% of the drug during 2 hr of dissolution in 0.1 N HCl, and the commercial tablets released no drug. In phosphate buffer (pH 6.8) both formulations released their drug content in 1 hr. Both formulations are, therefore, in compliance with the USP requirements for release from enteric coated dosage forms. The in vivo availability study in six beagle dogs revealed that the formulated enteric-coated beads filled in hard gelatin capsules had a 197.54% bioavailability relative to that of the commercial Voltaren tablets. The tablets showed a significantly lower (p < 0.05) area under curve for 0-8 hr (AUC0 8 hr) of 13.44 +/- 15.02 micrograms hr/ml compared to 26.55 +/- 5.19 micrograms hr/ml for the capsules. The capsules showed a nonsignificantly (p > 0.05) higher peak plasma concentration (Cmax) of 6.77 +/- 0.67 micrograms/ml compared to 5.88 +/- 7.38 micrograms/ml for the tablets. The time to reach peak (Tmax) values were 2 +/- 1.48 and 2.25 +/- 1.08 hr for the capsules and tablets, respectively. The capsules showed less interdog variability with respect to Cmax (CV% 34.6) and AUC (CV% 19.55) compared to CV% 79.9 and 111.76, respectively, for the commercial tablets. PMID- 9876512 TI - The effect of structural changes on swelling kinetics of polybasic/hydrophobic pH sensitive hydrogels. AB - The effect of pendent side-chain length and crosslinking agent concentration in methyl methacrylate/dimethylaminoethyl methacrylate as a polybasic/hydrophobic pH sensitive hydrogel was studied. Increasing both side-chain length and crosslinking agent concentration decreased the sharpness of response to pH and water-uptake capacity of the polymer. A case-II water transport mechanism and a nonlinear swelling kinetic was observed for the homologues of this hydrogel. PMID- 9876514 TI - Effects of drug concentration in inner aqueous phase and additives in oleaginous phase on release and bioavailability of isoniazid from multiple emulsion. AB - The effects of drug concentration in internal aqueous phase of stabilized w/o/w emulsion and additives in oleaginous phase on release characteristics of isoniazid were investigated. The release was significantly effected by both of the formulation variables. The release was enhanced initially with increasing concentration of drug in internal aqueous phase followed by a steady release at high concentration of isoniazid. The release declined substantially in the presence of aluminum tristearate, cetostearyl alcohol, and cholesterol, and it increased with egg lecithin and oleic acid in oily phase. The bioavailability was increased with a multiple-emulsion formulation. PMID- 9876513 TI - Brain drug delivery system bearing dopamine hydrochloride for effective management of parkinsonism. AB - Dopamine hydrochloride bearing positively charged small liposomes was prepared by sonicating the multilamellar vesicles. These vesicles were characterized for their physical attributes (shape, size, charge, drug entrapment efficiency, and drug leakage). The drug release kinetics from the liposomes were also studied and found to be Fickian-type diffusion. In vivo performance of the drug-entrapped liposomes was assessed by periodic measurement of drug- (chlorpromazine) induced catatonia in Sprague-Dowley rats. These results were compared with the plain dopamine HCl and levodopa preparations as well with the marketed formulation of levodopa containing carbidopa (Syndopa). These studies revealed that the dopamine can be effectively delivered to the brain by incorporating it into liposomes, and its degradation in circulation can also be protected. The results of liposomal formulation were found to be superior compared to plain levodopa as well as Syndopa. PMID- 9876515 TI - The role of mixed micelles in drug delivery. I. Solubilization. AB - The present study revealed that the solubilization of a poorly soluble drug, Amphotericin-B (AmB) can be improved by lipid-bile salt mixed micelles and it was also noted that mixed micellar systems are thermodynamically stable relative to micellar systems. It was observed that the degradation or precipitation of AmB was significantly reduced when stored at 8 degrees C. The mixed micellar systems composed of sodium desoxycholate with monoolein increased the solubilization of AmB by more than seven-fold as compared to nonmicellar systems. PMID- 9876516 TI - The effect of keratolytic agents on the permeability of three imidazole antimycotic drugs through the human nail. AB - The permeability of three imidazole antimycotics (miconazole nitrate, ketoconazole, and itraconazole) through the free edge of healthy human nail was evaluated in vitro using side-by-side diffusion cells. The influence of keratolytic substances (papain, urea, and salicylic acid) on the permeability of the antimycotics was also studied. The results suggested that the nail constituted an impermeable barrier for these antimycotics; it could be considered that the nail behaved as a hydrophilic gel membrane, through which drugs of low solubility could not permeate. The use of ethanol did not promote the passage of any of the antimycotic drugs. Although scanning electron microscopy indicated that the keratolytic substances had a significant effect on the nail surface (papain > salicylic acid > urea), the passage of the three antimycotics was not improved by pretreatment with salicylic acid alone (20% for 10 days), or by the application of the drug in a 40% urea solution. It was found that only the combined effects of papain (15% for 1 day) and salicylic acid (20% for 10 days) were capable of enhancing the permeability of the antimycotic. PMID- 9876517 TI - Simultaneous determination of ascorbic acid, pyridoxine hydrochloride, and tyrosine by derivative UV spectrophotometry. AB - A new UV spectrophotometric method for resolving a three-component drug mixture of ascorbic acid (I), pyridoxine hydrochloride (II), and tyrosine (III), on the basis of the first-derivative UV spectra and zero-crossing technique, is described. Beer's law was obeyed in the concentration range of 7.0-15.0, 0.25 1.0, and 8.0-40.0 micrograms/ml for I, II, and III, respectively. Lower limits of detection at the 95% confidence level were 2.88, 0.69, and 5.43 micrograms/ml, respectively. The advantages of the proposed method (speed, easy sample preparation, and lower cost per analysis) also include its application to the content uniformity and dissolution test of a three-component mixture of drugs. PMID- 9876518 TI - Enhancement of bioavailability of ketoprofen using dry elixir as a novel dosage form. AB - To enhance the dissolution rate and bioavailability of poorly water-soluble ketoprofen, a novel oral dosage form of ketoprofen, termed ketoprofen dry elixir, was developed by the spray-drying technique. Ketoprofen, dextrin, and sodium lauryl sulfate were dissolved in an ethanol-water mixture (20:25 w/w) and thereafter spray-dried to form the ketoprofen dry elixir. Comparative studies on the in vitro dissolution and in vivo adsorption of ketoprofen in the form of dry elixir and powder were carried out. Ketoprofen in the dry elixir completely dissolved within 5 min. On the other hand, only about 50.1% of ketoprofen powder alone dissolved during 60 min. The initial dissolution rate of ketoprofen in the dry elixir markedly increased in distilled water at 37 degrees C, becoming fourfold higher than that of ketoprofen powder alone. The maximal plasma concentration of ketoprofen (Cmax) and the area under the concentration-time curve from zero to 8 hr (AUC0-8 hr) after the oral administration of dry elixir increased about 3.2- (24.6 versus 7.6 micrograms/ml) and 2.2-(38.4 versus 17.3 micrograms hr/ml) fold compared with powder alone. It was obvious that ketoprofen dry elixir might be a useful solid dosage form to improve the dissolution rate and bioavailability of poorly water-soluble ketoprofen. PMID- 9876519 TI - Controlled drug delivery by biodegradable poly(ester) devices: different preparative approaches. AB - There has been extensive research on drug delivery by biodegradable polymeric devices since bioresorbable surgical sutures entered the market two decades ago. Among the different classes of biodegradable polymers, the thermoplastic aliphatic poly(esters) such as poly(lactide) (PLA), poly(glycolide) (PGA), and especially the copolymer of lactide and glycolide referred to as poly(lactide-co glycolide) (PLGA) have generated tremendous interest because of their excellent biocompatibility, biodegradability, and mechanical strength. They are easy to formulate into various devices for carrying a variety of drug classes such as vaccines, peptides, proteins, and micromolecules. Most importantly, they have been approved by the United States Food and Drug Administration (FDA) for drug delivery. This review presents different preparation techniques of various drug loaded PLGA devices, with special emphasis on preparing microparticles. Certain issues about other related biodegradable polyesters are discussed. PMID- 9876520 TI - Multiple-layer compression-coated tablets: formulation and humidity studies of novel chewable amoxicillin/clavulanate tablet formulations. AB - The purpose of this study was to produce novel multiple-layer, compression coated, chewable tablet formulations containing amoxicillin trihydrate, and clavulanic acid as potassium clavulanate, and to test in vitro dissolution characteristics and the effect of humidity stability compared to Augmentin chewable tablets as a reference. Double- and triple-layer tablets were manufactured on a laboratory scale by multiple-layer dry compression, and dissolution profiles of both active ingredients were determined. Tablets were subjected to stability evaluation in laboratory-scale humidity tanks maintained at constant humidity. Assay of content was determined by HPLC or UV spectroscopy. Physical characteristics of the powder mixture, such as angle of repose, and of tablets for hardness and friability, were also determined. Chewable tablets showed similar dissolution profiles in vitro for both active ingredients, compared to the marketed reference, Augmentin. The stability of clavulanic acid, but not amoxicillin, was increased in the novel triple or bilayer formulation. The tablets showed suitable friability, hardness, and angle of repose for starting materials to suggest that industrial scale-up is feasible. This approach to formulation of drugs containing multiple or moisture-sensitive ingredients has been shown to increase the stability of the central core drug without changing the dissolution pattern of the active ingredients. This formulation is expected to be bioequivalent in vivo based on these in vitro results. PMID- 9876521 TI - Bead compacts. I. Effect of compression on maintenance of polymer coat integrity in multilayered bead formulations. AB - Little information is available on the compactability of beads for oral sustained release dosage forms. It is known that polymer-coated beads may fuse together to produce a non-disintegrating controlled-release matrix tablet when compressed. This study evaluates the effect of compression on beads with multiple layers of polymer and drug coat, and the effect of cushioning excipients and compaction pressure on drug release from compressed bead formulations. The multilayered beads consist of several alternating layers of acetaminophen (APAP) and polymer coats (Aquacoat) with an outer layer of mannitol as a cushioning excipient. Percent drug release versus time profiles showed that the release of drug decreases from noncompacted beads as the amount and number of coatings increases, with only 43% of drug released in 24 hr for coated beads with 10 layers. It was shown that the compacted multilayered beads will disintegrate in gastrointestinal fluids, providing a useful drug release pattern. It was shown that beads of drug prepared by any method can be spray-layered with excipients such as Avicel and mannitol. Spray-layering of the cushioning excipient onto beads can provide an effective way to circumvent segregation issues associated with mixing of the polymer-coated beads and powdered or spherical/nonspherical cushioning excipients. Spray layering of the cushioning excipient can also provide excellent flow properties of the final formulation as visually observed in our experiments. Triple-layered caplets (TLC) were also prepared with outer layers of Avicel PH 101 or polyethylene oxide (PEO), and a center layer of polymer-coated beads. For TLC, the polymer coating on the beads fractured, and nondisintegrating matrix formulations were obtained with both caplet formulations. PMID- 9876522 TI - Thermal analysis as a screening technique in preformulation studies of picotamide solid dosage forms. AB - The potential compatibilities of several commonly used pharmaceutical excipients with picotamide were evaluated using differential scanning calorimetry (DSC). The effects of aging and of mechanical treatment (blending, grinding, or kneading) of samples were also evaluated. Hot-stage microscopy (HSM) and scanning electron microscopy (SEM) were used as complementary techniques to implement and assist in interpretation of the DSC results. DSC analysis evidenced a noticeable modification of drug thermal features in the mixtures with palmitic acid, stearic acid, stearyl alcohol, polyethylene glycol (PEG) 20,000, and sorbitol, but HSM analysis showed that the DSC behavior was mainly because of the drug dissolution in the melted excipient, which allowed the presence of important solid-solid interactions to be excluded. Compatibility with Mg stearate was also found, even if sample manipulation induced the partial conversion of Mg stearate in a pseudo polymorphic modification. Mechanical stress displayed an increased hygroscopicity of mixtures with glucose and lactose, as well as some solid-solid interactions with lactose and mannitol. PMID- 9876523 TI - Effect of cod-liver oil extract on the buccal permeation of ergotamine tartrate. AB - Ergotamine tartrate (ET) is used clinically in the treatment of migraines. However, the bioavailability of ET is rather poor following oral administration. Therefore, we tried to improve ET delivery using buccal administration. The purpose of this study was to investigate the characteristics of the permeation of ET through the hamster cheek pouch in vitro using a two-chamber diffusion cell, and to evaluate the effect of permeation enhancers on the transbuccal delivery of ET. Cod-liver oil extract (CLOE), polyoxyethylene hydrogenated castor oil (HCO 60), sodium glycocholate (GC), and sodium caprate (CA) were selected as premeation enhancers considering their low irritancy of the mucosa. When the enhancers were added to the donor cell at a 5% concentration each, the ET permeation rate markedly increased compared with that in a control not containing enhancer. Among these enhancers, CLOE exhibited the greatest effect. Because CLOE is composed of 16 kinds of fatty acids, the enhancement action of each of the major components was separately determined. As major fatty acids, palmitic acid, oleic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) were selected and their enhancing effects were studied. The enhancing effect of each fatty acid was significantly lower than that of CLOE. PMID- 9876524 TI - Influence of propellant composition on drug delivery from a pressurized metered dose inhaler. AB - Hydrofluoroalkanes (HFAs) are used to replace chlorofluorocarbons (CFCs) as non ozone-depleting propellants for pressurized metered-dose inhalers (pMDIs). HFA 134a and HFA 227 are used in combination to precisely manipulate the density and vapor pressure of pMDI formulations. The influence of propellant composition on the dose delivery characteristics of a suspension-based pMDI formulation was investigated. The results showed that as the density of the propellant blends approached the density of the suspended drug particles, the formulation became more physically stable and exhibited the most consistent dose delivery and greatest respirable fraction. The mass median aerodynamic diameter of the aerosolized particles contained in the emitted dose also was decreased by using propellant blends with higher vapor pressures. The performance of a suspension based pMDI formulation was optimized by varying the propellant composition using HFA 134a and HFA 227. PMID- 9876525 TI - Evaluation of two dextrose-based directly compressible excipients. AB - The objectives of this research were to evaluate the physical properties and compaction behavior of two dextrose-based directly compressed excipients. Anhydrous theophylline (10% w/w) was used as a drug model, Emdex and or Maltrin M510 (89.5% w/w) were used as diluent, and magnesium stearate (0.5% w/w) was used as lubricant. Direct compression and wet granulation methods were used for preparing the compacts. In general, the wet granulation method reduced the density of the mixture and consequently its flow rate compared to the mixture prepared only by solid-solid mixing. All formulations were compressed at four different compressional forces and at a target weight of 450 mg +/- 5%. Tablets obtained were different in physical properties and mechanical strength based on type of excipient used and methods of tablet preparation (direct compression versus wet granulation). Compacts prepared from Maltrin M510 had a longer disintegration time and slower drug release than compacts of the same composition but prepared with Emdex. Disintegration time and drug dissolution from tablets containing Maltrin M510 as diluent and prepared by wet granulation appeared to be controlled by a "gel" layer formation around the tablets and not by the tablets porosity. This study demonstrates that full characterization of excipients is needed because a different manufacturing process for the same excipients may produce differences in the pharmaceutical products. PMID- 9876526 TI - Different molecular weight chitosan microspheres: influence on drug loading and drug release. AB - Influence of chitosan molecular weight on drug loading and drug release of drug loaded chitosan microspheres was studied. Chitosans of 70,000 (LC), 750,000 (MC), and 2,000,000 (HC) molecular weight were employed alone or as mixtures (HC/LC 1:1 1:2 w/w). Ketoprofen (ket) was chosen as the model drug to be encapsulated. Microspheres characterized by different theoretical polymer/drug ratios were prepared (2:1, 1:1, 1:2 w/w). Satisfactory ket contents were obtained for all batches of chitosan microspheres with the theoretical polymer/drug ratio 1:2 w/w; microspheres made of HC/LC (1:2 w/w) were characterized by good drug content and encapsulation efficiency independent by polymer/drug ratio. Prepared chitosan microparticulate delivery systems can modulate ket release within 48 hr. Microspheres consisting of HC/LC (1:2 w/w) were the most suitable formulation in controlling drug release. PMID- 9876527 TI - Effect of Explotab on the tabletability of a poorly soluble drug. AB - The efficiency of a superdisintegrant (Explotab) in a direct-compression formulation containing a poorly water soluble drug (albumin tanate) at high dosage was investigated. An experimental design with two variables, applied pressure and concentration of Explotab, enabled its effects on the tableting and the mechanical properties of the final tablets to be determined. Differential scanning calorimetry was performed to study the interactions between drug and excipients. No incompatibility was found between drug-excipient mixtures prepared in the proportion 1:1 and in the corresponding formulation at room temperature and after 3 weeks at 50 degrees C. The concentration of Explotab has a positive effect on flow properties. Also, the effect of applied pressure and disintegrant content was found to be significant on all compressional parameters. At low applied pressures, the breaking strength was independent on Explotab concentration. However, at higher applied pressures, the maximum densification obtained with 10% Explotab produced a limited breaking strength lower than that at 0% concentration. The response surface shows a certain level of Explotab, around 7%, at which the disintegration time was the shortest. At this level, the surface response was independent of the applied pressure. In our study, the experimental design was a valuable tool used to establish the optimum manufacturing conditions. PMID- 9876528 TI - Fabrication of multiunit controlled-release phenylpropanolamine hydrochloride tablets. AB - Phenylpropanolamine hydrochloride (PPA) pellets were prepared in a fluidized-bed rotary granulator. Microcrystalline cellulose and distilled water were used as pelletization enhancer and binder, respectively. The pellets were coated with methacrylate ester copolymer (Eudragit RS 100) solution containing a 1:1 ratio mixture of triethyl citrate and castor oil as plasticizers. The addition of approximately 30% microcrystalline cellulose and 2% croscarmellose sodium to the 50% coated pellets produced fast disintegrating tablets. Dissolution profiles of both pellets and their respective matrix tablets were comparable and conformed to the USP dissolution requirement for PPA extended-release capsules. PMID- 9876529 TI - Lisinopril-lactose incompatibility. AB - A chemical interaction between lisinopril and lactose was demonstrated and quantified in dilute mixtures in the solid state. PMID- 9876530 TI - A mixture experiment approach for controlling the dissolution rate of a sustained release tablet. AB - Several sustained-release tablet formulations with acceptable pharmacokinetic properties were found to be unstable because of the effects of lactose. Because the pharmacokinetic properties were acceptable, an attempt was made at developing stable formulations that reproduced the in vitro drug release characteristics of the unstable formulations. Through the use of a statistically designed mixture experiment, alternative formulations were generated and tested for dissolution. The dissolution data collected in the mixture experiment were used to develop a statistical regression model for identifying formulations with dissolution rates equal to those of the unstable formulations. The form of the regression model was based on the Higuchi equation. The data analysis indicated that it is possible to generate dissolution profiles that reproduce those of the original formulations by adjusting the ratios of Methocel K4MCR Premium and Methocel K100MCR Premium and by replacing the detrimental lactose with calcium phosphate dibasic anhydrous. PMID- 9876531 TI - Studies comparing Kollicoat MAE 30 D with commercial cellulose derivatives for enteric coating on caffeine cores. AB - The products that are processed in aqueous form, such as Aqoat MF (suspension), Aquateric (pseudolatex), HP 55 (ammonia-based solution), and Kollicoat MAE 30 D (latex), were compared (in the form of spray dispersions, isolated films prepared from the dispersions, and caffeine-film-coated tablets with 5.5, 8.0, and 11.0 mg film/cm2) with one another and with ethanolic HP 55 S solution. The addition of pigments to all of the liquid preparations, with the exception of the ammoniacal solution of HP 55, led to a slight increase in pH. In each case, the viscosity of both solutions was well above that of the other formulations. The minimum film forming temperature was decidedly reduced by the addition of pigment. Kollicoat MAE was the undissolved film-former that had the smallest particle size and particle size distribution. The next smallest were those of Aqoat MF. The latex and the suspension were the only products that were sensitive to shear and heat. The isolated films did not display any tack. The strongest films and the films most impermeable to water vapor were obtained from solutions, and this can be ascribed to the fine distribution of the film-former. None of the isolated films showed signs of dissolving at pH 4.5. At pH 5.5, only the HP 55 was dissolved. This was because HP 55 was processed in ammonia-based solution; as a result of which, films that were not very resistant to gastric juice were obtained. The other formulations did not dissolve until the pH reached 6.0. As the pH rose, the rate of dissolution increased for all of the films. The permeability to protons was similar to that of caffeine-film-coated tablets to gastric juice. The resistance increased in the following sequence: HP 55 (ammonia-based) < Aquateric < Aqoat MF < HP 55 S (organic) and Kollicoat MAE. As a result of the temperature treatment and the rate of spraying, the production time on a 5-kg scale was twice as long for 5.5 mg Aqoat MF/cm2 as it was for Kollicoat MAE. This amount of film sufficed for Kollicoat MAE and HP 55 S solution to achieve adequate resistance to gastric juice. Aqoat MF did not attain the same resistance until a thickness of 11 mg film/cm2 was reached. Film tablets with Aquateric and ammonia-based HP 55 solution absorbed more than 20% of gastric juice at this film thickness. PMID- 9876532 TI - In vivo characteristics of injectable poly(DL-lactic acid) microspheres for long acting drug delivery. AB - Poly(DL-lactic acid) (PLA) microspheres containing testosterone (T) were prepared by the solvent evaporation process to evaluate their physical properties such as size distribution, shape, drug content, in vivo controlled drug release, pharmacological influences on the prostate gland in castrated rats, and histopathological findings of tissues surrounding the implants. The in vivo release of T from PLA microspheres containing 30 mg of drug obtained with chloroform was continued over a 6-week period. This effect is attributed to high dispersibility of T in the device when obtained with chloroform. Both serum drug levels and prostate gland weight recovery suggested the effects of a long-acting drug delivery system. The histopathological findings showed that the devices used were completely degraded 10 weeks after injection. PMID- 9876533 TI - Thermal and mechanical characterization of cellulose acetate phthalate films for pharmaceutical tablet coating: effect of humidity during measurements. AB - Films from a polymer used in pharmaceutical coating (cellulose acetate phthalate) were analyzed by thermomechanical techniques including dynamic mechanical thermal analysis (DMTA) and tensile tests. Emphasis was placed on relative humidity (RH) at the measurement site (as opposed to storage or conditioning RH). The films were plasticized with either triethyl citrate or diethyl phthalate. The results show that the films respond rapidly to changes in the environmental humidity. This in turn influences the data obtained from DMTA and tensile testing; thus, good control of the humidity is essential. Absorption isotherms have been obtained for the two types of films, and the results were interpreted in terms of the equilibrium moisture content which is determined by the polar nature of the plasticizer. This factor must be considered when formulating a film composition because moisture can, apart from its influence on mechanical properties, also speed the chemical degradation processes. When the stability of a film composition during storage is studied, the actual measurements on the films should preferably be done at 0% RH, irrespective of the storage RH. At 0% RH, the mechanical testing results will reflect the chemical stability correctly, without interference from plasticizing effect of absorbed water. A functionality test for films has been suggested: DMTA under isothermal conditions using either step or continuous humidity scans. Isothermal dynamic humidity scans on the DMTA were performed for the first time, and the utility of these measurements is discussed. PMID- 9876534 TI - Development of tablets for controlled joint release of nifedipine and atenolol. AB - Oral combinations of nifedipine and atenolol are widely used in the treatment of hypertension, proving particularly effective when the atenolol is released immediately and the nifedipine is released in a sustained manner. This work examined the potential of combining nifedipine and atenolol in a tablet, which would be easier to manufacture than currently available combined formulations. The results indicated that a 40:60 (w/w) nifedipine-atenolol mixture forms a eutectic melting at 140 degrees C. Nevertheless, both drugs were stable when incorporated in tablets elaborated using cellulose ethers as base excipients. Tablets prepared from atenolol-lactose granules and solid dispersions of nifedipine-hydroxypropylmethylcellulose (100 cP) had more adequate dissolution profiles than a more complex reference formulation in hard capsules. PMID- 9876535 TI - Preparation and testing of cyclosporine microsphere and solution formulations in the treatment of polyarthritis in rats. AB - We prepared a microencapsulated sustained-release formulation of cyclosporine A (CsA) and compared its efficacy to the solution formulation of cyclosporine A (Sandimmune, Sandoz) in an attempt to improve the treatment of rheumatoid arthritis. Microspheres containing cyclosporine were prepared with poly(lactic co glycolic acid) (PLGA), a polymer in the submicron particle range of 0.22-0.8 micron. Studies were carried out to determine uptake rates and mechanisms of lymphocyte inhibition mediated by macrophages containing CsA microspheres in vitro. The results of these studies were used to establish whether lower doses of the microencapsulated cyclosporine could be used in in vivo studies in the polyarthritic rat model for rheumatoid arthritis. In vitro dissolution testing revealed that CsA was released extremely slowly from microspheres for up to 48 hr (0.002%). Radiolabeled 3H CsA was incorporated into some PLGA microspheres or the microspheres were labeled using a 99mTc radioligand when needed, and radiolabeling efficiency was consistently above 50%. Uptake studies at various microsphere-to-macrophage ratios (1:1, 1:5, 1:10) were carried out using 99mTc radiolabeled microspheres and macrophages obtained from normal and polyarthritic rats. Normal macrophages behaved significantly differently from arthritic macrophages throughout the study. Arthritic macrophages cause increased amounts of CsA to be released (68% of the dose) into the culture medium past 24 hr compared to normal macrophages (48% of the dose). This factor may account for the significantly increased inhibition (68.2%) of mixed lymphocyte culture proliferation in the presence of arthritic macrophages containing CsA-loaded PLGA microspheres over normal macrophages (48.2%) that were pre-exposed to the same microsphere dose. The equivalent quantity of CsA as that contained in the microspheres when placed in solution or the same quantity of blank PLGA microspheres caused decreased levels of lymphocyte inhibition when compared to the effects of CsA microspheres in macrophages of normal cells, but significantly decreased levels of inhibition in arthritic cells. From the in vivo studies, it is evident that CsA microspheres, even at low dose levels, were highly effective in inhibiting polyarthritis in rats. PMID- 9876536 TI - Comparative evaluation of surfactant and hydrophobizing agent concentration in relation to the optimal particle size of metered-dose inhalers. AB - The aim of the present study was to formulate stabilized suspension-type metered dose inhalation aerosols, and to examine the connection between the stabilizing additives and the optimal particle size. For the stabilization of the suspended particles, hydrophilic- and hydrophobic-type additives were applied. Oleil oleate was selected as a hydrophilic anionic surfactant, and the hydrophobizing agent was dimethyl siloxane polymer. The effect of the amount of the applied hydrophilic and hydrophobic additives on the optimal particle size was modeled by a second-order polynomial equation fitted to the data gathered by a face-centered central composite statistical design. We found that if the proper type and amount of additives are selected, it is possible to acquire the therapeutically best composition. PMID- 9876537 TI - Influence of powder characteristics of bulk substance with pharmaceutical processing. AB - The physicochemical properties of crystals can vary with the crystallization procedure employed in their isolation and purification. Moreover, the success of any direct-tableting procedure is directly effected by the quality of the crystals used in this process. We examined the conventional crystallization method employed in the isolation and purification of octotiamine crystals, the active component of the pharmaceutical compound Neuvita. Our objective was to determine under what crystallization conditions (i.e., supersaturation ratio [pH], temperature, impeller speed) octotiamine crystals with excellent direct tableting potential could be obtained. Our results indicated that modifications in pH level (from 4.3 to 4.0), i.e., a reduction in the supersaturation ratio, and in impeller speed (from 100 to 78 rpm) are necessary to obtain octotiamine crystals with superior flowability and compressibility compared to the use of the conventional crystallization method. Thus, with these modifications in the conventional crystallization method, octotiamine crystals can be made that show dissolution rates similar to those of the conventionally made crystals, yet which can be manufactured into tablets using a simpler method (i.e., direct tableting). Also, the tableting powder made from the new crystal type proved to be less adhesive than the conventionally made crystal powder. This property attributed to the new crystal type will allow for more stable automated manufacturing than the conventional crystal type would allow. PMID- 9876539 TI - The use of beta-cyclodextrin as a pelletization agent in the extrusion/spheronization process. AB - The use of beta-cyclodextrin for the preparation of pellets by the extrusion/spheronization process is described for different formulations and processing conditions. Sieve analysis and friability tests were performed to assess the physical and technological characteristics of pellets. Satisfactory products were obtained with beta-cyclodextrin contents up to 90% by weight. PMID- 9876540 TI - Effect of menthol on permeability of an optically active and racemic propranolol across guinea pig skin. AB - The effect of menthol on the percutaneous penetration of S(-)-propranolol (SPL) and racemic form of propranolol (RSPL) was investigated in vitro using excised abdominal skin of guinea pig. In the presence of menthol, the permeability coefficient of SPL was high compared with that of RSPL. The enhancement factors for SPL and RSPL were 2.12 and 0.85, respectively. The lag times for SPL and RSPL were reduced considerably in the presence of menthol compared to those for control (without enhancer). The present findings suggest the enantio-selective permeation of SPL across the guinea pig skin in the presence of menthol. PMID- 9876538 TI - Evaluation of a sulfobutyl ether beta-cyclodextrin as a solubilizing/stabilizing agent for several drugs. AB - To evaluate the potential use of beta-cyclodextrin sulfobutyl ether, 7 sodium salt (SBE7-beta-CD) as a drug solubilizing and stabilizing agent, the solubilizing effects of SBE7-beta-CD on 22 different poorly water-soluble drugs were compared with those of intact beta-CD and heptakis-(2,6-di-O-methyl)-beta-CD (DMCD). SBE7-beta-CD was generally a more effective solubilizer for poorly water soluble drugs than was intact beta-CD, but SBE7-beta-CD was not as effective as DMCD. The effects of SBE7-beta-CD on the acid hydrolysis rate of prostaglandin I2, the alkaline hydrolysis rate of indomethacin, the dehydration of prostaglandin E1, and the isomerization of prostaglandin A1 were also investigated and compared to those for intact beta-CD, DMCD, and 2,3,6 partially methylated-beta-CD (PMCD). The stabilizing effects of SBE7-beta-CD on chemically unstable drugs were generally higher than those of other CDs. PMID- 9876541 TI - Preparation and evaluation of erythromycin fumarate--a new derivative of erythromycin. AB - Erythromycin fumarate, a new water-soluble derivative of erythromycin, was prepared and its physicochemical and biological properties were evaluated. The derivative also has considerable solubility in organic solvents. Its partition coefficient data in different organic solvent-water systems may indicate that it is well-distributed in various tissues in vivo. Antimicrobial potency in vitro of the derivative, 725 micrograms/mg, is much higher than that of the existing derivatives and its antimicrobial spectrum is comparable to that of the parent antibiotic. The LD50 value of the new derivative in mice intraperitoneally is 402.7 mg/kg. Results of this and the previous investigation on pharmacokinetics and protein binding indicate that erythromycin fumarate has high potential for possible clinical application and further investigation may be undertaken. PMID- 9876542 TI - Entrapment efficiency and initial release of phenylbutazone from nanocapsules prepared from different polyesters. AB - Several formulations of poly(epsilon-caprolactone) (PCL), poly(lactic acid) (PLA), and poly(lactic-co-glycolic acid) (PLGA) nanocapsules containing phenylbutazone were prepared according to the interfacial deposition technique. These formulations differed in the type of polymer used to form the shell of the nanocapsules. Analysis of particle size distribution and encapsulation efficiency of the nanocapsules revealed that the type and molecular weight of polyester used were the main factors influencing these properties. PLA had the highest encapsulation efficiency with the best reproducibility. From in vitro release studies, a small amount of drug release was observed at pH 7.4. However, in the gastric medium, an important burst effect occurred and was highest with the PLGAs and lowest with PCL, suggesting that drug release from these systems is affected by the type of polymer and the environmental conditions. The two formulations of phenylbutazone-loaded nanocapsules should be evaluated based on PCL and PLA in vivo in order to determine to what extent they are able to reduce the local side effects of this drug. PMID- 9876543 TI - Prolonged release of tegafur from S/O/W multiple emulsion. AB - To develop a prolonged and sustained release preparation, we prepared an albumin microsphere-in-oil-in-water emulsion (S/O/W) and examined sustained release from it in comparison with other control preparations such as water-in-oil (W/O) emulsions and microspheres in vitro and in vivo, respectively. Tegafur was used as a model drug. A microsphere-in-oil emulsion was prepared by adding albumin microspheres to soybean oil containing 20% Span 80. To prepare an S/O/W emulsion, the microsphere-in-oil emulsion was added into an aqueous solution of hydroxypropyl methylcellulose containing Pluronic F68. The mean particle size of the albumin microspheres was 3 microns, and the ratio of entrapment of tegafur into albumin microspheres was about 25%. In an in vitro release test, the t75 of the S/O/W emulsion was fourfold greater and in an in vivo release test the mean residence time of tegafur from the S/O/W emulsion was more than twofold that from a W/O emulsion or microsphere system. The mean residence time of 5-fluorouracil (5-FU) from an S/O/W emulsion was also greater than with other dosage forms. These results suggest the possible usefulness of an S/O/W emulsion for the sustained and prolonged release of tegafur. PMID- 9876544 TI - Physicochemical stability of crystalline sugars and their spray-dried forms: dependence upon relative humidity and suitability for use in powder inhalers. AB - Lactose, trehalose, sucrose, and mannitol were purchased in crystalline form and fractionated by sieving. Coarse (125-212 microns) and fine (44-74 microns) free flowing fractions were selected as typical of drug carriers in dry-powder inhalers. In addition, one batch of each sugar was spray-dried to form a respirable powder (> 50% [w/w], < 5 microns). Both fractions and the spray-dried powders were characterized before and after storage for 30 days at < 23%, 23%, 52%, 75% and 93% relative humidity (RH) at 25 degrees C. Moisture uptake was determined by thermogravimetric analysis (TGA) validated by Karl Fischer titration. Sieve fractions (before storage at different RHs) and spray-dried materials (before and after storage) were further characterized by differential scanning calorimetry (DSC) and x-ray powder diffraction (XRPD). All crystalline sieve fractions (except sucrose at 93% RH) were stable at 25 degrees C and showed insignificant moisture uptake when exposed to each relative humidity for 30 days. Sucrose dissolved in sorbed moisture at 93% RH. Spray-dried lactose, sucrose, and trehalose, which were collected in the amorphous form, showed moisture uptake, without recrystallization, when held for 30 days at 23% RH. These sugars recrystallized as sintered masses and became undispersible at > or = 52% RH. Spray-dried mannitol was apparently 100% crystalline when collected directly from the spray-dryer; it did not show humidity-induced changes. The physicochemical behavior of each sugar form is discussed as it relates to the sugar's suitability as a powder-inhaler excipient, with both conventional and protein drugs. PMID- 9876545 TI - Modeling the effects of hydroxypropylcellulose in acetaminophen tablet formulation. AB - The objective of the research described here was to develop a set of predictive models that would be used to show the performance of hydroxypropylcellulose as a pharmaceutical tablet binder. A statistically designed set of experiments was used to relate tablet formulation to functionality. It was found that the binder level affected both hardness and dissolution time. Useful predictive models were generated for tablet hardness and dissolution time as a function of the binder or binder-drug ratio. The optimal formulation can be predicted from this study, and will depend upon the combination of desired hardness and the dissolution time for a particular drug. PMID- 9876546 TI - Evaluation of a mucoadhesive buccal patch for delivery of peptides: in vitro screening of bioadhesion. AB - We have assessed the bioadhesive properties of several different mucoadhesive buccal patches. The patches consisted of custom coformulations of silicone polymers and Carbopol 974P. The contact angle of water was measured for each of the test formulations, using an ophthalmic shadow scope. The corresponding work of adhesion between the water and the patches (W1), and between the patches and freshly-excised rabbit buccal mucosa (W2) was then calculated, using a modification of Dupre's equation. The bioadhesive strength between the patches and excised rabbit buccal mucosa was also assessed. The results of the contact angle measurements indicated that the contact angle decreased with an increase in the amount of Carbopol in the formulation. Additionally, the calculated values of both W1 and W2 increased with an increase in the amount of Carbopol in the buccal patch formulations. A correlation (r not equal to 0.9808) was found between the measured contact angle and the calculated values for W2. The direct measurement of the force required to separate a buccal patch from excised rabbit buccal mucosa with the INSTRON demonstrated that the adhesive strength increased with an increase in the amount of Carbopol. This preliminary study has shown that the measurement of contact angles alone may provide a useful technique for estimating the work of adhesion, and may serve as a convenient and rapid screening procedure to identify potential mucoadhesive buccal-patch formulations. PMID- 9876547 TI - Preparation and characterization of oil-in-water type poly (D,L-lactic acid) microspheres containing testosterone enanthate. AB - Poly (D,L-lactic acid) (PLA) microspheres containing testosterone enanthate (ET) were prepared by using an oil-in-water (O/W) emulsion technique. The size distribution of the microspheres obtained could be explained by a log-normal distribution, and as a result, it was found that ET fully incorporates into microspheres even when the drug is loaded at up to 50%. On the other hand, the dissolution behavior of ET from microspheres was strongly dependent on particle size, suggesting that dissolution of the drug from microspheres can be easily controlled by controlling the preparative conditions. PMID- 9876548 TI - Surfactant dissolution and water solubilization in chlorine-free liquified gas propellants. AB - The initial water content of a group of 15 pharmaceutically and toxicologically acceptable surfactants showed a tendency to increase with the surfactant hydrophilic-lipophilic balance (HLB) value. Surfactant solubility was determined in chlorine-free "alternative propellants" (n-butane, propane, dimethyl ether [DME], 1,1,1,2-tetrafluoroethane (HFA-134a), and 1,1,1,2,3,3,3-heptafluoropropane [HFA-227ea], and trichloromonofluoromethane [CFC-11] in the absence of cosolvents such as ethanol. Water-soluble surfactants such as Carbowax, Sentry, PEG 300, Tween 20, and Brij 30, with high HLB values showed appreciable solubility in HFA 134a and HFA-227ea. In systems containing > or = 80% propellant by weight, each single-phase propellant-surfactant blend was screened for its ability to solubilize iodine and dissolve or solubilize water with increasing surfactant concentration. This screening was performed to investigate the possibility of formulating high-volatility, single-phase systems with increased polarity and solvency from these conventional excipients and vehicles. Ternary-phase diagrams show the regions of apparent single and multiple phase behavior in each system. Despite the increased polarity of the hydrofluoroalkanes (HFAs), appreciable water solubility was seen only with these surfactants in DME and in the hydrocarbons (HCs) n-butane and propane. PMID- 9876549 TI - Development of diclofenac sodium controlled release solid dispersion powders and capsules by freeze drying technique using ethylcellulose and chitosan as carriers. AB - Controlled-release, solid dispersions of diclofenac sodium (DS) were prepared by freeze-drying technique, using ethylcellulose (EC) and chitosan (CS) as single and combined carriers. Factorial design was applied as an experimental design to study the main and interactive effects of EC and CS on drug dissolution from the controlled release solid dispersion. All DS solid dispersions showed slower drug dissolution than did DS powder. The equations of dissolution parameters as functions of EC and CS contents were established through multiple regression. The contour plots of the established equations were constructed. The 10:(2.4 + 0.05) DS:(EC + CS) solid dispersion was prepared and developed into a capsule dosage from, using lactose as diluent. The effect on capsule dissolution of a disintegrant, sodium starch glycolate (Explotab), in concentrations of 2%, 5%, and 8% was studied. The solid-dispersion capsule containing 5% Explotab was found to provide the most similar dissolution profile to the one obtained with the 10:(2.4 + 0.05) DS:(EC + CS) solid-dispersion powder. The dissolutions of the 10:(2.4 + 0.05) solid-dispersion powder and capsules were closer to a first-order model than to a zero-order or diffusion control model. PMID- 9876551 TI - The use of particle characteristics to elucidate mix homogeneity in binary powder blends. AB - A series of binary powder blends comprising microcrystalline cellulose (Avicel PH101), alpha-lactose monohydrate, or anhydrous theophylline were prepared in order to investigate the ability of particle-characteristics measurements to express the homogeneity of the resulting mixture. It is postulated that fundamental physical characteristics, such as particle-specific surface area, true density, and size distribution, can be used to quantitatively ascertain mix homogeneity in routine pharmaceutical blending operations. PMID- 9876550 TI - Pharmacokinetic and bioequivalent study of a generic Metoprolol tablet preparation. AB - A study was conducted to compare the in vivo bioavailability of a generic metoprolol tablet preparation (Metoprolol) with that of the innovator product, Betaloc. Both preparations have a labeled dose of 100 mg metoprolol tartrate. Twelve healthy adult male volunteers participated in the study, which was conducted according to a standard two-way crossover design with a washout period of 1 week. The bioavailability was compared using the total area under the plasma level versus time curve (AUC0-infinity), peak plasma concentration (Cmax), and time to reach peak plasma concentration (Tmax). No statistically significant difference was observed between the logarithmically transformed AUC0-infinity values or the logarithmically transformed Cmax values of the two preparations. However, a statistically significant difference was observed between the Tmax values, but may not be therapeutically significant or important. Moreover, the 90% confidence interval (CI) for the ratio of the logarithmically transformed AUC0-infinity values of Metoprolol over those of Betaloc was calculated to be between 0.94 and 1.02, while that of Cmax was between 0.98 and 1.01, both of which are within the acceptable limit of 0.80-1.25. From the data obtained, it was also observed that a high proportion of our volunteers of Asian origin appeared to be poor metabolizers of metoprolol, which was consistent with what had been observed in our previous study of another preparation of metoprolol. PMID- 9876552 TI - An investigation of the influence of the core material properties on the compression and properties of dry-coated tablets. AB - The effect of core material properties on the process of compression and physical properties of compression-coated tablets were investigated using microcrystalline cellulose as the coating material (mantle). Three model core materials: borosilicate glass, silicone rubber, and steel, each of different Young's modulus, were selected to give a range of core mechanical properties. Coated tablets were prepared using a single-punch press, with facilities for monitoring the compression cycle and analysis of data using the Heckel (1) equation. This analysis showed a considerable effect of different core materials on the compression process, (i) as an unanchored core, and (ii) due to core material type with differing Young's modulus. PMID- 9876553 TI - Chitosan: a unique polysaccharide for drug delivery. AB - The aim of this review is to give an insight into the many potential applications of chitosan as a pharmaceutical drug carrier. The first part of this review concerns the principal uses of chitosan as an excipient in oral formulations (particularly as a direct tableting agent) and as a vehicle for parenteral drug delivery devices. The use of chitosan to manufacture sustained-release systems deliverable by other routes (nasal, ophthalmic, transdermal, and implantable devices) is discussed in the second part. PMID- 9876554 TI - Assessment of pharmaceutical quality of furosemide tablets from multinational markets. AB - This report describes results of a collaborative study in which samples of the 40 mg strength of furosemide tablets were evaluated following a common protocol based on British (BP), European (Ph. Eur.), and US Pharmacopoeial (USP) specifications. Several tests, including identification, uniformity of mass, and dissolution, were performed. In total, excluding Lasix lots, results for 162 lots obtained from 115 manufacturers or suppliers were submitted. Also, 23 laboratories identified and submitted data for 34 lots of Lasix products available in their countries. There were no reported abnormalities in the physical test requirements of the products analyzed. The summaries (n, mean, and 95% CI) of the assay results for the "standard sample" (a common sample), Lasix lots from participating countries, and for all other furosemide products, respectively, are as follows: 30, 99.8%, 96-104; 33, 100.0%, 94-106; and 162, 99.6, 94-105. About half (approximately 62%) of the reported uniformity of mass results based on tablet weights were in the range 150-175 mg/tablet. However, there appears to be notable variability in tablet weights that would result in significant differences in the ratios (0.14 to 0.40) of active ingredient to excipient. The reported disintegration times ranged from 0 (instantaneous) to 18 min, with most less than 1 min. The drug dissolution testing was conducted with phosphate buffer at pH 5.8 (USP recommended). Another test was conducted with acetate buffer at pH 4.6 (noncompendial). There appears to be remarkable similarity in overall percentage of drug release from the three types of products (standard sample, Lasix lots, and other products). Although apparently there is a very wide spread in dissolution characteristics of the products tested, the analyses of variance did not detect differences among the products tested and, to this extent, would not indicate differences in bioavailability characteristics for most of these products. It is observed that about 20-38% of the variability in dissolution testing is not product related (i.e., it is from the dissolution testing itself), while the remaining 62-80% variability is product related (manufacturing, formulation, etc). The results of this multinational collaborative study showed that most of the furosemide products available in different countries met the required pharmaceutical quality standards, including drug-release characteristics. Based on an extensive statistical analysis, the main concern from the study was that the high variability in drug dissolution testing would require wide tolerance standards (e.g., pharmacopoeial standards). This may result in lack of needed discriminating ability of the test in revealing the impacts of formulation and manufacturing changes on in vitro, and perhaps in vivo, drug-release characteristics. PMID- 9876555 TI - Hydrophilic-lipophilic drug carrier systems of bead cellulose and isopropyl myristate. AB - Solid carrier systems of bead cellulose (BC) and isopropyl myristate (IPM) as a lipophilic excipient showed high release rates for low prednisolone and high IPM content and decreasing rates for increasing drug and decreasing IPM content. The release of griseofulvin was decreased by increasing IPM content. The release was controlled by the solubility of the drugs in IPM and water, the crystalline state of the drugs, and the weight ratio of the components and was varied in a wide range by change of the prescriptions. The products were prepared according to a dispersion-coevaporation process and were received as flowable powders consisting of spherical porous particles. IPM with dispersed drug was incorporated into the pores of the beads and also precipitated on the bead surface. Depending on the ratio of IPM to drug, more or less crystalline drug particles were suspended in the liquid IPM film or the drug was amorphous dissolved. Investigations of wettability and water uptake gave hints to more lipophilic properties. The advantage of the coprecipitates was the combination of a hydrophilic carrier and a lipophilic excipient as a flowable system for controlled release. PMID- 9876556 TI - Advantages of impact testing over hardness testing in determining physical integrity of tablets. AB - An investigation of four different tablet strength tests was carried out on four different placebo formulations (differing in Avicel: Pharmatose ratios). The results analysis compared fatigue failure, work of failure, and impact failure to diametrical compression measurements (hardness). The impact results clearly show how different formulations can have the same hardness, yet their impact resistance can vary by as much as 200%. The impact test used in this work and other tests described are useful in tablet development to understand, compare, and mitigate tablet breakage during subsequent unit operations. PMID- 9876557 TI - Characterization of cellulose acetate phthalate (CAP). AB - Cellulose acetate phthalate (CAP) is a commonly used enteric coating polymer. CAP powder has been studied by various methods to determine characteristics that have an influence on its functionality. While some of the parameters are well known, such as free-acid content and substituent composition, new methods have been developed to examine them. Other characteristics, such as the molecular mass distribution, have not been reported earlier. Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), and thermal analysis have also been performed on fresh samples, as well as samples stored under various temperature and humidity conditions. Humidity is by far a more critical storage parameter than temperature, although the two act in conjunction; high humidity is more deleterious to the functionality of the polymer than high temperature. Functionality in this case is taken to be determined by the substituents and by the molecular mass distribution. Mass-average molecular mass of a number of batches of the polymer has been measured and ranges around 48 kg/mol with a degree of polydispersity of 1.6. A method to perform a rough estimation of the molecular mass of CAP has also been suggested based on knowledge of the substituent content. It may be possible to use the values of and obtained here for any other batch of the same viscosity grade of CAP. NMR has been employed to determine the fraction substituents in the polymer. However, an attempt to obtain the pattern of substitution of the CAP molecule by NMR was unsuccessful. Glass transition temperatures of CAP samples were measured. However, this characteristic of the polymer is judged not as sensitive to the loss of substituents as the molecular mass. Thermal treatment of the polymer in oxygen and inert atmospheres gave slightly different degradation products. PMID- 9876558 TI - Influence of temperature on the emitted dose of an oral metered dose inhaler. AB - The performance of metered dose inhalers is critical for the efficient delivery of drugs to the intended site of deposition in the respiratory tract. The temperature at which metered dose inhaler products are used by patients may influence the physicochemical characteristics of the emitted dose. Product performance characteristics of a metered dose inhaler containing beclomethasone dipropionate and oleic acid in a blend of chlorofluorocarbon propellants, Freon 11 and Freon-12, were determined by cascade impaction analysis and dose delivery through the valve after the metering chamber was loaded and actuated at 4 degrees C, 23 degrees C, and 40 degrees C. The dose delivered from the valve was not affected by the temperature at which the metering chamber was loaded and actuated. The mass median aerodynamic particle size of the emitted aerosol decreased and the percentage respirable fraction increased as the temperature was increased. The geometric standard deviation of the particle size distribution was not significantly affected by the temperature at which the metering chamber was loaded and actuated. The temperature at which a metered dose inhaler is used by a patient may influence the amount of drug that is potentially respirable; therefore, the dose expected to be delivered and the corresponding therapeutic effect may also be affected. PMID- 9876559 TI - The composition of NF-defined emulsifiers: sorbitan monolaurate, monopalmitate, monostearate, monooleate, polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80. AB - Using the analytical constants for sorbitan monolaurate, monopalmitate, monostearate, and monooleate given in the National Formulary (NF), calculations were carried out that indicated that these emulsifiers are esters of sorbitol mono- and dianhydrides. Contrary to the NF description, no significant amount of sorbitol ester was calculated to be present, in agreement with recent experimental findings. Further calculations were made using the NF-defined analytical constants of polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80, which indicate that these emulsifiers are esters primarily of polyoxyethylated sorbitol monoanhydride (i.e., sorbitan), with lesser quantities of polyoxyethylated esters of sorbitol dianhydride. Since all hydroxyl groups of the polysorbates are primary, random distribution of acyl groups on the available hydroxyls can be assumed, and the likely composition of these emulsifiers can be calculated. The most abundant compounds appear to be polyoxyethylated sorbitan mono-, di-, and triesters. Although the polysorbates are stated to contain 20 moles of ethylene oxide per mole of ester, the oxyethylene contents stated in the Food Chemicals Codex reveal that somewhat more than 20 moles of ethylene oxide are combined. PMID- 9876560 TI - Simultaneous optimization of wet granulation process involving factor of drug content dependency on granule size. AB - Computer optimization technique was applied to the simultaneous optimization of wet granulation process by a high-speed mixer granulator. Four pharmaceutical properties, including yield, drug content uniformity, geometrical mean diameter of granules, and uniformity of granule size, were selected to evaluate the quality of the granules. In particular, dependence of drug content uniformity on granule size was investigated using two model drugs, ascorbic acid and ethenzamide. An appreciable dependence of ascorbic acid content on granule size was not observed in model formulations. On the other hand, ethenzamide was contained more in small-size granules, and its content was decreased with an increase in amounts of hydroxypropyl cellulose (HPC-L; used as a binder) and binder solution. These observations suggested that drug content uniformity is influenced not only by drug solubility in the binder solution, but also by the use of HPC-L. A simultaneous optimal point incorporating four pharmaceutical properties was obtained using the generalized distance function. The experimental values of the four response variables obtained in newly prepared granules were found to correspond well with the predicted values of both granules containing ascorbic acid and ethenzamide. These results suggested that computer optimization would benefit the wet granulation process even if drug content segregation was involved in the process. Further, data obtained from computer optimization, in particular the contour diagram, will be valuable in the process validation. PMID- 9876561 TI - In vitro performance of floating sustained-release capsule of verapamil. AB - Capsules filled with mixtures of verapamil, hydroxypropoxyl cellulose (HPC), and effervescent are proposed to provide floating sustained release over 10 hr. The effects of weight filled in the capsule, amount of HPC, and the addition of effervescent on the dissolution kinetics are studied. The conventional capsules were filled with different amounts and weights of the mixtures of verapamil, HPC, and effervescent. The release of verapamil from the capsules followed the Higuchi release model. However, when effervescent was added, a zero-order drug release was observed after the burst phase. The conventional capsule, when filled with active ingredients, polymers, and effervescent, can achieve a zero-order release system. Entrapped air was considered as a barrier to diffusion and matrix relaxation in drug release. PMID- 9876562 TI - Effect of the formulation parameters on the characteristics of pellets. AB - Pelletization is increasingly applied currently for the preparation of solid oral controlled-release dosage forms. The production of the particles, which are regular in shape and size, can be achieved with the application of the proper polymer auxiliary materials and new pharmaceutical technological methods (extrusion, spheronization). Regularity in shape and size, attained by the optimization of several production parameters, can promote the coating procedure. Under optimal conditions, particles were prepared for coating in a high-shear mixer, which is used to produce uniform particles. The effect of the rotating speed of the applied chopper and the amount of microcrystalline cellulose in the composition on the physical characteristics of the pellets was modeled by a second-order polynomial equation fitted to the data gathered by a face-centered central composite statistical design. PMID- 9876563 TI - Dissolution profiles of flurbiprofen in phospholipid solid dispersions. AB - This study is concerned with the development of a solid dispersion formulation of flurbiprofen (FLP) and phospholipid (PL) with improved dissolution characteristics. The FLP powders were blended with PL to produce FLP-PL physical mixtures or made into solid dispersions with PL by the solvent method. The FLP exhibited significantly improved dissolution rates in PL coprecipitate (coppt) compared to the physical mixtures or FLP alone. The dissolution studies suggested that less than a 20:1 ratio of FLP to PL was required to disperse FLP completely in the carrier. The coppt yielded a ninefold greater initial dissolution rate. Also, the total amount dissolved after 60 min was twofold greater at a 10:1 ratio of FLP to L-(-dimyristoyl phosphatidylglycerol (DMPG). Similar results were observed with a ratio as low as 20:1 (FLP:DMPG). Increasing the DMPG content did not increase the rate to any significant extent. Thus, a small PL:FLP ratio improved the dissolution to a significant level. Thus, an FLP:PL dispersion may have the clinical advantages of quick release and excellent bioavailability. PMID- 9876564 TI - Measurement of electrostatic charge decay in pharmaceutical powders and polymer materials used in dry powder inhaler devices. AB - The electrostatic charge generated on drug/excipient particles during the formulation, manufacture, and use of pharmaceutical dry powder inhaler (DPI) devices may significantly affect the performance of such devices. An experimental investigation has been undertaken of charge accumulation and decay on compacts of selected powders (lactose and salbutamol sulfate) and a device material (polyvinyl chloride, PVC) used during the formulation, manufacture, and use of DPIs. Significant differences in charge acquisition and decay for the three materials have been demonstrated after charging using a corona electrode. PVC acquired the highest charge, which decayed rapidly in 30 min toward the value prior to exposure to corona. Lactose and salbutamol acquired similar charge values, which decayed to zero after 30 min for lactose, whereas salbutamol retained a significant charge after 120 min. The significant differences in charging propensity among drug, excipient, and device materials may have relevance in DPI formulation, manufacture, and use. PMID- 9876565 TI - Particle size determination of a flocculated suspension using a light-scattering particle size analyzer. AB - Microscopy is a useful and direct method for measuring the particle size of a suspension because, in addition to the particle size and size distribution, it provides visual detection of the shape and state of aggregation of the particles in the suspension. However, this method suffers from the shortcomings of being tedious and time consuming. In this study, a light-scattering particle size analyzer was used to determine the particle size and size distribution of a flocculated suspension. The sonication of the sample prior to and during measurement was found to be critical in ensuring that data are representative of the size distribution of the primary particles of the suspension. The light scattering results were further confirmed by data generated using a polarized light microscope equipped with an image analyzer. PMID- 9876566 TI - Evaluation of guar gum in the preparation of sustained-release matrix tablets. AB - Polymeric hydrophilic matrices are widely used for controlled-release preparations. The process of drug release is controlled by matrix swelling or polymer dissolution. It has been shown that the swelling of guar gum is affected by concentration of drug and viscosity grade of the polymer. This study examines the mechanism of behavior of guar gum in a polymer-drug matrix. The swelling action of guar gum, in turn, is controlled by the rate of water uptake into the matrices. An inverse relationship exists between the drug concentration in the gel and matrix swelling. This implies that guar gum swelling is one of the factors affecting drug release. The swelling behavior of guar gum is therefore useful in predicting drug release. PMID- 9876567 TI - In vitro evaluation of dissolution properties and degradation products of omeprazole in enteric-coated pellets. AB - This report describes results of an in vitro study in which capsules containing omeprazole in enteric-coated pellets from different Brazilian manufacturers were evaluated. The original product was the reference in comparison to three similar products (A, B, and C). Samples were submitted to severe conditions (40 degrees C and 75% relative humidity during 120 days), and the tests performed were the omeprazole content, the percentage of omeprazole dissolved from the pellets, and the amount of H 238/85, its main degradation product. The data obtained suggest that these products could not be considered interchangeable. Differences in physical and physicochemical properties of products A, B, and C indicated that they did not maintain the required stability and that bioavailability might be affected by the poor dissolution of omeprazole from the pellets. PMID- 9876568 TI - Preliminary evaluation of shilajit as a suspending agent in antacid suspensions. AB - The efficacy of shilajit, a gummy exudate of the plant Styrax officinalis Linn (Family: Styraceae), was evaluated as a suspending agent for the formulation of antacid preparations. Shilajit produced effects on sedimentation volume similar to those produced by sodium carboxymethyl cellulose (CMC), but at lower concentrations. It induced better flocculation with a moderate increase in viscosity compared to CMC. It did not interfere with the acid-consuming capacity of the suspensions. PMID- 9876569 TI - Preparation techniques and mechanisms of formation of biodegradable nanoparticles from preformed polymers. AB - The techniques available to prepare biodegradable nanoparticles (nanospheres and nanocapsules) from preformed polymers are reviewed. Although there is abundant literature on this topic, only a few focus on the thorough analysis of preparative procedures. In particular, four techniques are discussed in terms of their technological advantages and drawbacks: emulsification evaporation, solvent displacement, salting-out, and emulsification diffusion. The proposed mechanism of nanoparticle formation for each technique is described from a physicochemical perspective. The effects of preparative variables on nanoparticle size and drug entrapment efficiency are also discussed. PMID- 9876570 TI - Bioabsorbable polymers for implantable therapeutic systems. AB - For a long time, subcutaneous implantable drug pellets using nondegradable polymers have been used for long-term, continuous drug administration. The procedure requires surgical implantation and removal of the drug-containing devices or polymeric matrices, which has a significant negative impact on the acceptability of the product candidate. In addition, the release profile from such devices is neither constant nor readily controlled in terms of precision of rate of release and duration of action. These facts have led to the research and development of novel, controllable, nonirritating, noncarcinogenic, biocompatible, and bioabsorbable drug delivery systems for overcoming the drawbacks of nondegradable implantable pellets for prolonged continuous release. Biodegradable implantable systems release the drug over a long period of time with simultaneous or subsequent degradation in the tissue of the polymer to harmless constituents, thus avoiding removal once the therapy is complete. This approach has considerably improved patient acceptability and patient compliance. Various bioabsorbable polymers have been evaluated for controlled implantable drug delivery, including hydrogels, copolymers of polylactic and polyglycolic acids, polylactic acid, poly(orthoesters), polyanhydrides, poly(E-caprolactone), and polyurethanes. Their characteristics have been studied using a variety of drugs, like anticancer agents, hormone agonists and antagonists, nonsteroidal anti-inflammatory agents, neuroleptics, contraceptives, and others. The present paper describes the current research on implantable therapeutic systems, the bioabsorbable polymers, and the biologically active agents being used in this approach. PMID- 9876571 TI - Coatings for controlled-release drug delivery systems. PMID- 9876572 TI - The application of population pharmacokinetics to the drug development process. AB - Population pharmacokinetics is playing an increasing role in clinical drug development. An overview of the population approach, including software and the advantages and limitations of the approach compared to the traditional approach to pharmacokinetic studies, is given. This paper also documents how the area has evolved over the past 15 years and addresses some of the issues that have arisen over the design and conduct of population studies. Finally, some alternative applications of the population approach are given for areas other than clinical drug development. PMID- 9876573 TI - Applications of fiber-optic evanescent wave spectroscopy. AB - The evanescent wave (EW) component of light propagated via fiber-optic wave guides can be used to both sense and transmit information regarding the immediate environment of the fiber's surface. In this article, an outline of the theoretical and practical aspects of this emerging methodology is given, as well as a discussion of the advantages, disadvantages, and limitations of the technique. Examples are given of how EW spectroscopy may be used in the analysis of pharmaceutical systems. Evaluation of attributes of components of EW spectroscopy allows prediction of the future for this rapidly evolving area of photonics. PMID- 9876574 TI - Scientific and regulatory aspects of nutraceutical products in the United States. AB - Within the past decade, the so-called health food industry has experienced a tremendous growth such that these products are commonplace in most community pharmacies within the United States. These products have since been defined, by those in the industry, as nutraceuticals. Passage of the Nutrition Labeling and Education Act of 1990 and the Dietary Supplement Health and Education Act of 1994 has had a significant impact on the information available to the consumers to enable them to make educated decisions when "self-medicating" with this class of products. This paper is an attempt to summarize the current state of scientific and regulatory issues that are relevant to nutraceutical products. PMID- 9876576 TI - Critical dissolution tests of oral systems based on statistically designed experiments. II. In vitro optimization of screened variables on ER-coated spheres for the establishment of an in vitro/in vivo correlation. AB - The study was designed to optimize the effects of the screened in vitro dissolution variables agitation, temperature, osmolality, and polarity on the release of the neuroleptic compound remoxipride from extended release coated spheres. The variables were varied independently by means of a fractional factorial design. The in vitro tests were performed with the Basket method (USP). The polarity and the osmolality of the medium had significant effects on the dissolution rate of remoxipride. A statistical model was calculated based on the obtained dissolution in vitro. The model was then used to predict the in vitro conditions that most closely correlated with the dissolution rate of remoxipride in vivo, after administration of the formulation to 16 volunteers. The predicted in vitro conditions were experimentally verified, and an excellent association with the in vivo behavior of the formulation was found. Validation of the optimal in vitro conditions was performed on another batch of the formulation. The dissolution profile obtained showed a significant association with the corresponding dissolution profile in vivo. The use of statistically designed experiments in the development of critical dissolution tests for the establishment of in vitro/in vivo correlations seems to be a useful working approach, and supports further application to other oral solid systems. PMID- 9876577 TI - Therapeutic effect and pharmacokinetics of ketotifen transdermal delivery system. AB - Ketotifen transdermal delivery systems were prepared using polyisobutylene, liquid paraffin, and fatty acid. In vitro skin penetration studies were conducted in Franz diffusion cells using excised porcine skin to determine the skin permeation rates of ketotifen patches. A trend of increased skin penetration of ketotifen was observed as the amount of liquid paraffin in the patch was increased. In addition, we found that lauric acid was a suitable enhancer for percutaneous absorption of ketotifen. Challenge tests were performed in guinea pigs to determine the therapeutic effect of the delivery systems for the inhibition of anaphylactic shock using varied concentrations of chicken ovum albumin as sensitizer. Our results showed that compared with the treatment of intramuscular administration, the skin patch was more effective and produced higher survival rates. The pharmacokinetics of the ketotifen patch were determined by applying the skin patch to the dorsal skin of rabbits. The plasma levels were maintained constant (42.5-36.4 ng/ml) from 9 to 30 hr. From our study, the prepared ketotifen patch may further be developed for the treatment or prevention of allergic asthma. PMID- 9876578 TI - Simultaneous determination of pseudoephidrine HCl (PSE) and terfanidine (TER) from formulations by reversed-phase ion pair high-performance liquid chromatography (RP-HPLC). AB - A new simple, precise, rapid, and selective reversed-phase ion pair high performance liquid chromatography (RP-HPLC) method has been developed for the simultaneous determination of pseudoephidrine (PSE) and terfanidine (TER) from tablets using 60:15:25 acetontrile:methanol:water (v/v) containing 2.9 g sodium lauryl sulfate/liter, pH adjusted to 3.1 using phosphoric acid as a mobile phase and C18 Spherisorb ODS 2 (3 microns, 5 cm x 4.6 mm i.d.) as stationary phase. Detection was carried out using a UV detector at 254 nm. A constant flow of 1.0 ml/min was maintained throughout the analysis. Retention times for PSE and TER were 1.90 and 7.35 min, respectively. Linearity range and percentage recoveries for PSE and TER were 24-1200 and 12-600 micrograms/ml, and 100.01 and 100.4%, respectively. PMID- 9876579 TI - Study of standard tablet formulation based on fluidized-bed granulation. AB - In this study, acetaminophen, ascorbic acid, and ethenzamide were selected as model drugs for tableting granules. Agitation and fluidized-bed granulation were carried out at three drug contents of 30, 50, and 70%. Compared with agitation granulation, granules made by fluidized-bed granulation showed superior compressibility with wide formulation allowance for drug type and amount. Fluidized-bed granulation resulted in less granule hardness and greater plastic deformability. The granules had considerable compactness and for tablets containing 70% ethenzamide, prolonged disintegration and dissolution times were noted. These are typical features of granules produced by fluidized-bed granulation. PMID- 9876580 TI - Development of ointment formulations prepared with Achyrocline satureioides spray dried extracts. AB - Achyrocline satureioides spray-dried extracts, prepared with colloidal silicon dioxide, microcrystalline cellulose + colloidal silicon dioxide (1:1), and beta cyclodextrin + colloidal silicon dioxide (1:1), were incorporated in a glyceryl monostearate base. The influence of the spray-drying adjuvants on the formulations' physical characteristics, such as spreading properties, oil indexes, viscosities, and the pH determination, were evaluated. The results indicated that the adjuvants influenced the ointments' physical parameters at different levels, although all of them maintained their plastic flow and presented antithixotropic behavior. The presence of colloidal silicon dioxide alone, in the dried extract, imparted the lowest oil index value and an intermediary spreading area to the ointment. The colloidal silicon dioxide content reduction and the substitution of part of it by beta-cyclodextrin or microcrystalline cellulose enhanced the ointments' oil index values, while the best spreading area was reached by the ointment prepared with the spray-dried extract containing colloidal silicon dioxide and microcrystalline cellulose. PMID- 9876581 TI - Development of a new silicon-based transdermal system. I. Study of silicone elastomers and effect of liquid ingredients. AB - The polarities of four elastomers made of silicon oligomers of different viscosities were investigated by measuring the uptake of swelled solvent in different polarity solvents after 24, 48, and 72 hr of treatment. The solvent uptake provides a good characterization for the polarity of the inside of the matrix. On the basis of the results the oligomers were divided into two groups. The same method has been used for the investigation of the change of the inner polarity of samples containing various amounts of polar and apolar liquid ingredients. It was shown that the polarity of the inside of the matrix is related to the swelling properties. The ingredients used changed the matrix framework, which was also influenced by the type of oligomers. The solvent uptake shows large dependence on the timing of the measurement. Measurements carried out after 48 hr of polymer production showed constant solvent uptakes, indicating that the final structure of the polymer was formed. PMID- 9876582 TI - Skin healing preparations: compared in vitro diffusion of the active ingredients. AB - We have developed skin healing preparations based on propolis extract:ointments, emulsions, and transparent oil-water (TOW) gels. The formulations were optimized in terms of macroscopic characteristics including spreadability and penetrability, and limpidity and isotropy for the TOW gels. We describe here the results of a study of in vitro diffusion of the active ingredients, flavonoids, and phenolic acids, from some of the preparations we developed. These results show the influence of the dosage form and the choice of excipients on the release of the active product. The least satisfactory results were obtained for hydrophobic ointments and o/w emulsions. The strongest diffusion was obtained with a TOW gel associating isopropyl palmitate, Eumulgin B3, and Cetiol HE. PMID- 9876583 TI - Influence of plasticizers on tableting properties of polymers. AB - This study relates tablet formation with relaxation properties of two polymers on the basis of the stress-deformation curve. The mechanical properties of the polymers were varied by changing tableting temperature, adding varying amounts of plasticizer, and incorporating a monomer with plasticizer effect on the polymer chain. The crucial parameter appeared to be the difference between the glass transition temperature and the tableting temperature. This temperature difference was found to determine the amount of energy stored during densification. The energy is manifested as the stress relaxation propensity of the material. Large stress relaxation yields porous and consequently weak tablets. At a low temperature difference (i.e., tableting temperature is much lower than the glass transition temperature), the amount of stored energy is large. An increase in tableting temperature, or a decrease in glass transition temperature, yields a decrease in stored energy as a result of a decrease in yield strength. Consequently, production of less porous and stronger tablet is possible. However, if the tableting temperature is higher than the glass transition temperature, the stress relaxation propensity of the deformed polymers is extremely high because the elastic modulus of the materials is low under these circumstances. This results is porous and even capped tablets. From the data it is concluded that, independent of the type of polymer and the method of plasticizing, compaction at a temperature of about 20 K under the glass transition temperature yields circumstances for which the amount of stored energy has a minimum. Consequently, tablet porosity has a minimum and tablet strength has a maximum. These circumstances are created by changing both the tableting temperature and the glass transition temperature of the powder. PMID- 9876584 TI - Rotary tangential spray technique for aqueous film coating of indobufen pellets. AB - Indobufen pellets, previously prepared by direct pelletization employing the rotary tangential spray fluidized bed (Rotoprocessor), were coated using the same equipment setup as for the film coating process. Different amounts of the aqueous based ethylcellulose dispersion (Aquacoat) were easily applied on the active cores of different compositions with no spray blocking or pellet sticking. The coating process was not influenced by changing the ethylcellulose (Aquacoat ECD 30) formulation with soft acrylic resin (Eudragit NE 30D) or by adding water soluble polymers (Pharmacoat 606 and PEG 6000) to the coating dispersions. Simple setup, flexibility of operation, and short manufacturing times were the advantageous features of the tangential spray equipment. These benefits were verified during the pellet coating process. The composition of the cores and membranes were confirmed as the determining parameters with respect to the performances in terms of the drug release rate. The morphological and functional results clearly demonstrated the tangential spray rotary system as a promising one-step technique for the preparation of indobufen prolonged-release multiple unit dosage forms. PMID- 9876585 TI - Light-scattering method in particle size analysis of parenteral emulsions. AB - The use of a light-scattering particle size distribution analyzer has been shown to be a convenient method for characterizing the particle size distribution of parenteral emulsions. However, the concentrations of the samples used were found to have a major impact on the particle size distribution results, particularly for samples with a mean particle size smaller than 0.2 micron. An increase in sample concentration caused a shift to smaller particle sizes as a result of multiple scattering. The blue-light (tungsten lamp) intensity, instead of the He Ne laser integrity, should be used to control sample concentration within the optimal range for measurement. PMID- 9876586 TI - Development and in vitro evaluation of a buccoadhesive pindolol tablet formulation. AB - Controlled-release buccoadhesive tablets containing pindolol were prepared and evaluated in order to achieve constant plasma concentrations during the treatment of chronic hypertension and to improve the bioavailability of pindolol by the avoidance of hepatic first-pass metabolism. The formulations were tested for weight, hardness, friability, content uniformity, swelling rate, bioadhesive force, and drug release rate values. Carbopol 934 and NaCMC were used as bioadhesive polymers and Methocel K4M, Methocel K15M, and HPC were added as matrix-forming polymers. PMID- 9876587 TI - A stable multiple emulsion system bearing isoniazid: preparation and characterization. AB - Multiple emulsions with an oily liquid membrane (w/o/w) bearing isoniazid were prepared by an improved 2 x 2 step emulsification technique. Both of the interfaces of the liquid membrane were stabilized by using microcrystalline cellulose (MCC) in external as well as internal aqueous phases. The emulsions were characterized for droplet size, percent formation of multiple emulsion, release rate effect of Tween-80 in external phase, phase volume ratio on release, and stability during aging at various storage conditions. The droplet size was small and yield of multiple emulsion was fairly good. The increasing concentration of MCC in either internal or external phase increased the droplet size. The system holds promise in tuberculosis therapy. PMID- 9876588 TI - Solubilization and interaction of sulindac with polyvinylpyrrolidone K30 in the solid state and in aqueous solution. AB - In this report the interactions of sulindac with polyvinylpyrrolidone K30 (PVP K30), both in the solid state and in aqueous solution, have been investigated. Solid dispersions of sulindac with PVP K30 were prepared by the solvent method in ethanol from various drug-to-polymer weight ratios. X-ray powder diffraction and differential scanning calorimetry have shown that PVP inhibits the crystallization of sulindac. The stabilization of the noncrystalline state of sulindac was shown by x-ray diffractometry after a 1-year storage. There was a considerable increase in the release rate of the drug when the polymer content was increased and the intrinsic dissolution rate values of these systems were calculated. From the UV spectra a bathochromic shift and a well-defined isosbestic point were observed at pH 2 and 6, which confirmed an interaction between the drug and the polymer in solution. Moreover, the apparent solubility of sulindac has been modified as a function of the polymer concentrations. The binding process between the drug and PVP was exothermic from the stability constant values at 25, 30, and 37 degrees C at pH 2. PMID- 9876589 TI - Solubilization and interaction of sulindac with beta-cyclodextrin in the solid state and in aqueous solution. AB - A sulindac-beta-cyclodextrin complex was obtained by the coprecipitation method. Kneaded solids and physical mixtures were also prepared. The complex was shown by x-ray powder diffraction to be noncrystalline whereas pure drug and any of the other sulindac-beta-CD system were crystalline. the endothermic peak of sulindac due to the fusion of drug disappeared in DSC thermograms for the coprecipitate product, which confirmed the interaction between sulindac and beta-CD in the solid state. After a 1-year storage drug crystals could not be observed by x-ray diffractometry, which indicated that the complex formed was stable. The complex showed the fastest dissolution rate which might be attributed to the high-energy noncrystalline state and the inclusion complex formation in solution. UV spectra were modified and the apparent solubility of the drug increased with the addition of beta-CD, which confirmed the interaction between sulindac and the ligand in solution. The apparent stability constant, K1:1, for the complex at pH 2 and 25, 30, and 37 degrees C was 340, 220, and 160 M-1, respectively, which confirmed the influence of temperature on the complex stability. The value of K1:1 at pH 6 and 25 degrees C was 139 M-1, which indicated that the complex is formed easier with the non-ionized sulindac. The enthalpy change, delta H degree, showed that the binding process is exothermic. PMID- 9876590 TI - Hypoglycemic effect of oral insulin in diabetic rabbits using pH-dependent coated capsules containing sodium salicylate without and with sodium cholate. AB - The hypoglycemic effect of oral insulin (20 U and 40 U) capsules coated with a pH dependent soluble polymer (Eudragit S100) and containing sodium salicylate (50 mg) without and with sodium cholate (50 mg) was studied in alloxan-hyperglycemic rabbits and compared with that of s.c. insulin injection (20 U). The capsules containing 20 U insulin + sodium salicylate (50 mg) produced a significant reduction in plasma glucose level to 82 and 73% of initial values at 2 and 3 hr after administration, respectively. The blood glucose level slowly returned to normal values at 5 hr. The AUC0-5 hr was 73.7 +/- 43.5 mg.hr/dl compared to 242 +/- 70.5 mg.hr/dl for insulin (20 U, s.c.) with a relative hypoglycemia of 30.4%. A higher dose of oral insulin (40 U) + sodium salicylate (50 mg) was more effective in reducing plasma glucose level which steadily decreased and reached 56% of the initial value by 5 hr (AUC0-5 hr = 132 +/- 41.5 mg.hr/dl and relative hypoglycemia = 27.3%). Sodium cholate (50 mg), however, slightly improved sodium salicylate effect producing an AUC0-5 hr of 139 +/- 37.3 mg.hr/dl with relative hypoglycemia of 28.7%. The relative hypoglycemia of pH-dependent coated capsules reached in the present experiment is the highest found so far. PMID- 9876591 TI - Extension of the International Conference on Harmonization Tripartite Guideline for Stability Testing of New Drug Substances and Products to countries of climatic zones III and IV. AB - The International Conference on Harmonization (ICH) Tripartite Guideline "Stability Testing of New Drug Substances and Products" sets out the stability testing requirements for a registration application within the three areas of the European Union, Japan, and the United States. These areas are assigned to climatic zone II with the storage condition of 25 degrees C/60% RH. This paper describes the adjustments that are necessary to extend the guideline to countries of climatic zones III and IV. At first storage conditions were derived with 30 degrees C/35% RH for climatic zone III and 30 degrees C/70% RH for climatic zone IV. Both conditions contain a margin of safety compared to calculated and measured data in warehouses. Furthermore, they cover the extreme temperatures above 30 and 40 degrees C which may arise in these climatic zones. Six months at 40 degrees C/75% RH was fixed as storage condition for accelerated testing to assess organoleptic and physicochemical test criteria and to make predictions for chemical stability. The predictive factor is 3.3 for 30 degrees C (6 months at 40 degrees C corresponds to 20 months at 30 degrees C). Extreme temperatures which may arise during shipment are covered by the results of stress investigations (e.g., 3 months at 50 degrees C). The next adjustment is necessary for the selection of the packaging containers. They must reflect the requirements for solid, semisolid, and liquid dosage forms caused by the two storage conditions. In the evaluation the temperature difference of 10 degrees C (40-30 degrees C) instead of 15 degrees C has to be considered, which limits the preliminary shelf lives in critical cases to 18 months instead of 24 months. Finally, statement/labeling must reflect the storage conditions. All of the other basic principles for the drug substances and drug products, such as selection and number of batches, test criteria, test procedures, specifications, testing frequency, and storage period can be applied without any change. PMID- 9876592 TI - Formulation and in vitro evaluation of polymeric films of diltiazem hydrochloride and indomethacin for transdermal administration. AB - Ethylcellulose-polyvinyl pyrrolidone films containing diltiazem hydrochloride and indomethacin were evaluated for their potential drug delivery at a controlled rate, using rat skin, to select a suitable formulation for the development of transdermal drug delivery systems. The influence of film composition, initial drug concentration, and film thickness on the in vitro drug release rate as well as drug permeation through rat abdominal skin were studied. Drug release studies were carried out employing the paddle over disk method and drug permeation through full thickness of the rat abdominal skin was tested using a modified Franz diffusion cell fastened with O-ring. The drug content of the film decreased at an apparent first-order rate, whereas the quantity of drug released was proportional to the square root of time. The release rates of both drugs increased linearly with increasing drug concentration and polyvinyl pyrrolidone fraction in the film, but was found to be independent of film thickness. The increase in release rate may be due to leaching of hydrophilic fraction of the film former which resulted in the formation of pores. It was also observed that the release of drugs from the films followed a diffusion-controlled model at low drug concentrations. A burst effect was observed initially, however, at high drug loading levels. This may be due to rapid dissolution of the surface drug followed by the diffusion of drug through the polymer network in the film. The in vitro skin permeation profiles showed increased flux values with increase of initial drug concentration in the film and also with the concentration of polyvinyl pyrrolidone. From this study, it is concluded that the films composed of ethylcellulose:polyvinyl pyrrolidone:diltiazem hydrochloride (8:2:2) and ethylcellulose:polyvinyl pyrrolidone:indomethacin (8:2:3) should be selected for the development of transdermal drug delivery systems, using a suitable adhesive layer and backing membrane, for potential therapeutic use. PMID- 9876593 TI - Possibilities of conveying a cationic drug in Carbomer hydrogels. AB - A drug with cationic characteristics such as procaine can be conveyed in a Carbomer hydrogel in two different ways: (i) in the form of salt in solution in the aqueous phase, and (ii) in the base form salified with the same polymer. Introduction of the drug into the hydrogel with different concentrations of polymer produced, in both cases, a reduction in viscosity in relation to drug concentration. The gels with procaine salified with the polymer showed greater viscosity. The drug release rate, in general, diminished with the increase in polymer concentration. Nevertheless, when this concentration was maintained, there was no variation in release rate when the viscosity produced as a consequence of drug concentration was changed. Gels with procaine salified with the carboxyvinylic polymer had a faster release rate than those with procaine in the hydrochloride form dissolved in the aqueous phase. These results have also been confirmed by a simulated absorption test. PMID- 9876594 TI - Compatibility evaluation of metered-dose inhaler valve elastomers with tetrafluoroethane (P134a), a non-CFC propellant. AB - Compatibility of propellants, excipients, and solvents with the components of the valve greatly influences performance of metered-dose inhalers (MDIs). Ozone friendly hydrofluoroalkane propellant 134a has potential for use as a chlorofluorocarbon (CFC) replacement. No suitable replacement for propellant 11 and 114 has yet been found and the problems arising from this may be overcome by use of ethanol as a solvent. In this study, compatibility of MDI valve elastomers Dowty Nitrile 0117, White Buna, and Type 674 (B) with P134a placebo formulations having different concentrations of ethanol was investigated. The results indicate that formulations containing no ethanol adversely affected the functioning of the valves. Higher concentrations of ethanol improved valve performance, but showed increased leakage. Physical characteristics of the valve elastomers evaluated by determining swelling caused after exposure to the P134a placebo formulations exhibited increased swelling with increasing concentrations of ethanol in the formulation. PMID- 9876595 TI - Optimization of the processing of matrix pellets based on the combination of waxes and starch using experimental design. AB - An experimental design was used in order to optimize the one-step production process of matrix pellets based on the combination of waxes and starch. The parameters tested were the impeller speed (x1) and the mixing time (x2). Ibuprofen and theophylline were used as model drugs at a concentration of 60 and 70% (w/w), respectively. The 0.8-1.25 mm yield fraction of the matrix pellets was evaluated as the response factor Y. A quadratic equation was fitted to the experimental data and used to predict the response factor Y of the theophylline and the ibuprofen. The contour plots of both formulations revealed a flat and therefore rugged region from the upper left to the lower right of the domain investigated. The energy input into the system during the production process controlled the pellet growth, the impeller speed having a greater impact on the energy input compared to the mixing time. PMID- 9876596 TI - Alternative solvent-free preparation methods for felodipine surface solid dispersions. AB - Surface solid dispersions were prepared via physical mixture and were either heated in a vacuum dryer or in a microwave oven for different periods of time. The physical state of felodipine in solid dispersions was studied using differential scanning calorimetry and x-ray powder diffractometry. USP paddle method was used for felodipine dissolution studies. The use of vacuum or microwave energy led to a significant improvement of felodipine dissolution which was caused partly by the amorphous state of felodipine and a large surface area of amorphous silicon dioxide. PMID- 9876597 TI - Cyclodextrin solubilization of ETH-615, a zwitterionic drug. AB - Therapeutic usefulness of many zwitterionic drugs is hampered by their very low aqueous solubility. The purpose of this work was to investigate the effects of cyclodextrins on the solubility of the zwitterionic drug ETH-615, the role that charge might play in the cyclodextrin complexation, and the influence of polymers and ion-pairing agents on the cyclodextrin solubilization. The effects of five different beta-cyclodextrin derivatives were evaluated, i.e., the anionic beta cyclodextrin sulfobutyl ether sodium salt and carboxymethyl-beta-cyclodextrin sodium salt, the uncharged 2-hydroxypropyl-beta-cyclodextrin and randomly methylated beta-cyclodextrin, and the cationic 2-hydroxy-3-trimethyl ammoniopropyl-beta-cyclodextrin. The uncharged cyclodextrins had much larger solubilizing effect on ETH-615 than the charged ones. However, due to the highly polar zwitterionic structure of ETH-615 the stability constants of its cyclodextrin complexes were several orders of magnitude smaller than those commonly observed for uncharged lipophilic compounds. Cyclodextrin solubilization of ETH-615 was enhanced by water-soluble polymers and ion-pairing agents. PMID- 9876598 TI - Critical dissolution tests of oral systems based on statistically designed experiments. III. In vitro/in vivo correlation for multiple-unit capsules of paracetamol based on PLS modeling. AB - The main aims of the present study were to establish an in vitro/in vivo correlation for multiple-unit capsules of paracetamol by means of statistical prediction models and to investigate the effect of a number of in vitro variables on the discussion rate of paracetamol from the formulation. A fractional factorial screening design was used to investigate the effects of the variables agitation, pH, osmolality, viscosity, and the presence of bile salt on the dissolution rate of paracetamol. The effects were evaluated in two separate partial least-squares models, in which the responses were expressed as the cumulative percentage of paracetamol dissolved at specified time-points (model I) and as the shape (beta) and scale (eta) parameters according to the Weibull function (model II). It was concluded that agitation and viscosity had significant effects on the dissolution rate of paracetamol. Statistical models based on the responses from models I and II were then used to predict the in vitro conditions most closely correlated with the in vitro dissolution of paracetamol after administration of the formulation to 10 healthy volunteers. The predicted optimal in vitro conditions were similar for the two models and not too far from what is expected from the gastrointestinal tract. The experimental verification of the in vitro conditions showed that both models were equally good, and contributed to high degrees of correlation with the in vivo dissolution behavior of the formulation during 9 hr. The relationships obtained when plotting the percentage dissolved in vitro versus in vivo were y = 1.1x (r2 = 0.98) and y = 1.1x (r2 = 0.94) for models I and II, respectively. Based on these results, it is difficult to state a preference for one of the models. Finally, the use of statistical prediction models to develop critical in vitro tests is a successful approach in the establishment of associations between dissolution behavior in vitro and in vivo for oral extended-release systems. PMID- 9876599 TI - Effects of inorganic salts on solubilization of estriol in an aqueous solution of poly(ethylene oxide)/poly(propylene oxide)/poly(ethylene oxide) triblock copolymer. AB - The effects of inorganic salts on the solubilization of estriol was studied in an aqueous solution of a poly(ethylene oxide)/poly(propylene oxide)/poly(ethylene oxide) triblock copolymer (Pluronic L-64). The solubility of estriol increased with increasing Pluronic L-64 and salt concentrations. Furthermore, two clear transition points corresponding to the polymolecular micelle formation and a pronounced change in micellar structure of Pluronic L-64 were found on the solubility curves. The effectiveness of inorganic salts for increasing solubility of estriol followed approximately the lyotropic or Hofmeister series, except Li+. PMID- 9876600 TI - Improved dissolution rate of indomethacin by adsorbents. AB - Samples of indomethacin and kaolin or microcrystalline cellulose (Avicel) were prepared by solvent deposition or simple blending methods. Dissolution rates of these samples were studied. The surface adsorption of indomethacin on the studied adsorbents was shown to improve the dissolution rate of the drug in water. The solvent-deposited samples of indomethacin on kaolin or Avicel in the ratio 1:4 released 25% of the drug at 34 or 60 min, respectively (t25%), while 25% of the pure drug was released at 140 min. Meanwhile, the t25% of the corresponding drug adsorbent simple blends were 108 and 110 min, respectively. The effect of addition of polyvinyl pyrrolidone (PVP) as a third component to indomethacin adsorbent was studied and showed further improvement in in vitro availability of the drug-kaolin adsorbents. PMID- 9876601 TI - In vitro release of amoxycillin from lipophilic suppositories. AB - The in vitro release characteristics of amoxycillin from different lipophilic suppository bases were investigated using the USP rotating basket method. Suppositories containing 250 mg amoxycillin were prepared in theobroma oil and in the semi-synthetic bases Witepsol W35, Suppocire A32, Novata BD, and Novata 299. Both freshly prepared and 1-month-old suppositories were tested. Analysis of amoxycillin was performed using a validated high-performance liquid chromatographic (HPLC) technique. Release profiles differed significantly between bases, with the greatest amount of amoxycillin being released from both newly made and 1-month-old Novata BD bases (87.57 +/- 8.18 and 99.66 +/- 6.63%, respectively), and the lowest amount released from the newly manufactured theobroma suppositories (8.82 +/- 0.75%) and the 1-month-old Suppocire A32 suppositories (7.78 +/- 0.27%). PMID- 9876602 TI - Investigations on the influence of the type of extruder for pelletization by extrusion-spheronization. I. Extrusion behavior of formulations. AB - Three different extruders (the Alexanderwerk gravity feed roll extruder, the Gabler axial, single-screw extruder, and the NICA radial-screw extruder) were compared for their suitability for different placebo formulations and for fenoldopam pellets. A fourth extruder, the experimental ram extruder, was also included in some of the comparisons. Evaluation of the extrusion behavior of the three extruders showed differences as well as similarities among them, depending on the composition of the formulation. Although the NICA and Alexanderwerk units extruded all formulations successfully, the Gabler extruder failed to do so at a content of > 60% of soluble ingredients, such as lactose or mannitol. The extrudate surface improved for all extruders with an increase in water content of formulations, but was generally smoother for the Gabler than for the NICA or the Alexanderwerk units. A formulation with colloidal Avicel as spheronization aid showed an identical extrusion behavior for all of the investigated extruders. Of the three extruders, the Gabler unit showed the highest heat generation during extrusion, especially when extruding formulations with a low water content or high contents of soluble excipients. However, when the loss of water during extrusion or spheronization for various formulations was compared, only a two-way ANOVA test on the differences between the water content after extrusion and after spheronization showed a statistically significant difference between the Alexanderwerk or NICA and the Gabler extruder. The two-way ANOVA also proved that this difference is significant only for some formulations, e.g., lactose + Avicel PH 101 formulations, but not for Avicel PH 101 formulations. PMID- 9876603 TI - Investigations on the influence of the type of extruder for pelletization by extrusion-spheronization. II. Sphere characteristics. AB - Three different extruders, the Alexanderwerk gravity-feed roll extruder, the Gabler axial, single-screw extruder, and the NICA radial-screw extruder, were compared for their suitability for different placebo formulations and for fenoldopam pellets. A fourth extruder, the experimental ram extruder, was also included in some of the comparisons. The successful spheronization of extrudates from each of these extruders requires the correct water content. This water content, however, is different for each of the formulations and for each extruder. Generally, the Gabler unit required the highest amounts of water for a successful spheronization, yielding > or = 90% between 710 and 1250 microns. The NICA unit needed much less water for the same formulation, and the Alexanderwerk unit required even less water than the NICA unit. Pellet sphericity was also strongly dependent on the correct water content of formulations, but was generally better for pellets produced with the Alexanderwerk or NICA units. A two way ANOVA test for the individual formulations showed a significant difference in the mean particle size of batches produced with the NICA or the Alexanderwerk and the Gabler extruder. No significant differences could be found between any of the Alexanderwerk or NICA batches. Both extruders showed a linear dependence of the mean particle size on the water content of formulations, but the Gabler extruder showed an almost unchanged particle size over a wide range of water contents, provided that the formulation could be extruded successfully. Batches that were extruded on the NICA unit showed a significantly lower bulk density than comparable Alexanderwerk or Gabler batches. Comparing the true density of pellets, we found that significant differences could only be stated for Avicel PH 101 + water batches and only for the NICA/Gabler interaction. True density increased for all three extruders with increasing amounts of soluble components and with increasing water content. The NICA batches also exhibited a significant difference of the Hausner factor from the other two extruders, but no differences could be found in the friability of pellets. PMID- 9876604 TI - Small-scale characterization of wet powder masses suitable for extrusion spheronization. AB - The method of compresso-rheology with an Instron 5567 was used for flow assessment of wet powder mass in order to improve its formulation. In our experiments, the method was efficient for selection of the excipient (Avicel CL611) able to improve the extrusion behavior of the high-dose wet powder mass. The method also allowed the determination of the minima and maxima of the wetting agent volume necessary to identify the correct moisture content for extrusion (20%). The results were not discriminative for the choice of the Avicel CL611 amount in the formulae even if an average amount necessary to improve extrudability of the active ingredient could have been estimated at about 10%. Nevertheless, this method appeared to be a rapid and easy small-scale method for studying wet powder mass, cause only a few grams of solids are required, this rheometer should prove useful in formulation research. PMID- 9876605 TI - Effects of pH, electric current, and enzyme inhibitors on iontophoresis of delta sleep-inducing peptide. AB - Delta sleep-inducing peptide (DSIP), a peptide of nine amino acid residues, was used as a model drug to investigate the effects of pH, electric current, and enzyme inhibitors on the transdermal iontophoretic delivery of peptide drugs. DSIP was fairly stable in pH 4-9 buffer solutions but was cleaved by the skin enzymes during iontophoretic delivery. Enzyme inhibitors, such as o phenanthroline, ethylene-diaminetetraacetic acid (EDTA), dilucine, and sodium deoxycholate, could inhibit the degradation of DSIP to a certain extent in the skin homogenate. Our results showed that metalloproteases were probably more important enzymes for DSIP hydrolysis. By using 0.2 mM o-phenanthroline in the iontophoretic delivery of DSIP at pH 4, we were able to significantly enhance the penetration of DSIP. The flux was about eight times as much as control (without o phenanthroline) at pH 7.4. PMID- 9876607 TI - A study of the changes during heating of paracetamol. AB - The orthorhombic form of paracetamol has been shown to exhibit greater compressibility and faster dissolution than the monoclinic form. The orthorhombic form is produced by melting of monoclinic crystals of paracetamol followed by cooling at specific rates. Cooling rate, although a very important factor, is not the only factor influencing the formation of either of the two morphs. To study the cooling rate required for production of form II, paracetamol samples were melted in a differential scanning calorimeter, cooled at three specific rates, and melted again. In all of the samples, cooling resulted in the glassy form followed by recrystallization and the melting of form II. On the hot-stage microscope both forms were produced in one sample. Standardizing conditions for prediction of the resulting form remains a problem. There seems to be a great deal of overlap of the two forms' transition phases, which would make it difficult to force the crystallization of one form by keeping the solution or melt at a specific temperature. The thermal behavior of paracetamol during the heating and cooling phases must be understood in order to manipulate the process. A video camera mounted on a hot-stage microscope was used to follow the changes during heating and cooling of both forms. Nucleation, crystal growth, habit transformation, sublimation, and the final melt are shown on snap shots taken from the video. PMID- 9876606 TI - Effect of chemical structure on the release of certain propionic acid derivatives from their dosage forms. AB - The aim of this study was to investigate the relationship between the chemical structure and release properties of certain drug products. Propionic acid derivatives were used as a model. These include ibuprofen (I), ketoprofen (K), tiaprofenic acid (T), flurbiprofen (F), and naproxen (N). They are all aryl derivatives of propionic acid and differ only in the aryl group. Such an aryl group may be either isobutylphenyl, benzoylphenyl, benzoylthienyl, fluorobiphenyl, or methoxynaphthyl group in I, K, T, F, and N, respectively. Three dosage forms were selected for this study: capsules, suppositories, and creams. The release of propionic acid derivatives from the capsules and suppositories decreased in the order ibuprofen > tiaprofenic acid > ketoprofen > flurbiprofen > naproxen, and for the creams the release decreased in the order ibuprofen > tiaprofenic acid > flurbiprofen > ketoprofen > naproxen. The difference in drug release in the first case was attributed to the difference in the chain length, and in the creams which are composed of two phases, the partition coefficient was found to affect the drug release. The molecular weight of the drug had no effect on the release. The drug release from different dosage forms was not affected after 1 month storage. PMID- 9876608 TI - Preparation, characterization, and dissolution studies of ibuprofen solid dispersions using polyethylene glycol (PEG), talc, and PEG-talc as dispersion carriers. AB - Solid dispersions of ibuprofen (IBF) were prepared by solvent evaporation method using polyethylene glycol 10000 (PEG), talc, and PEG-talc as dispersion carriers. The drug-carrier(s) interactions in the solid state were investigated using scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and x-ray diffraction analysis. Interactions in the solution were studied by performing dissolution experiments. No important and well-defined chemical interaction was found between the ingredients. The increase in the IBF dissolution rate from the solid dispersions with the carriers used in this study could be attributed to several factors such as improved wettability, local solubilization, and drug particle size reduction. PMID- 9876609 TI - Preparation, characterization, and evaluation of physicochemical properties of different crystalline forms of ibuprofen. AB - Different crystal forms of ibuprofen (IBF) were prepared using methods such as cooling hot solution of the drug and precipitation of crystals from the drug solution. Scanning electron microscopy (SEM), differential scanning calorimetry (DSC), melting point, x-ray powder diffractometry, infrared absorption spectroscopy (IR), and in vitro dissolution rate and stability studies were conducted to investigate various characteristics of different crystalline forms of the drug. Methods of preparation and nature of the solvents used in this study were found to have greater roles in changing the physicochemical properties of IBF. PMID- 9876610 TI - Product development studies on the tablet formulation of ibuprofen to improve bioavailability. AB - To improve the bioavailability of ibuprofen, a thorough preformulation trial was undertaken. As a part of these studies, particle size of the drug, solubility-pH profile, and pH-partition coefficient profile were studied. The probability of improving solubility by solid dispersion technique was also investigated. The effects of different binding agents and incorporation of various proportions of sodium lauryl sulfate on the release rate of ibuprofen were also studied. The in vitro release profiles of the developed tablets showed superiority over the popular marketed tablets. PMID- 9876611 TI - Evaluation of hydrogel-based controlled-release niacin tablets. AB - Matrix-based controlled-release niacin tablets were formulated using guar gum. The effect on the in vitro dissolution profile was examined using variable guar gum content in the formulation. It was observed that the dissolution profile declined with the increase in the guar gum content in the tablet. The in vitro dissolution profile was also observed under different pH conditions and there was no marked change. The moisture content of granules also did not cause any considerable change in dissolution profile. Three different strength Niacin controlled-release tablets, including 500 mg, were prepared and it was found that applying the same variables of different guar gum content and moisture content of granules resulted in a very insignificant change in the in vitro dissolution profile. The experimental formulation compared well with commercial products and met the proposed standards for controlled-release products. PMID- 9876612 TI - Identification of the mebendazole polymorphic form present in raw materials and tablets available in South Africa. AB - A preformulation study of four different raw materials of mebendazole showed that three samples were polymorph C and the other polymorph A, or a mixture of form A and B. X-ray powder diffractometry and infrared spectroscopy indicated that this powder could be form B, but powder dissolution, for which a much slower dissolution was obtained, suggests polymorph A. Literature prescribes the use of polymorph C pharmaceutically, but generic manufacturers should be aware that forms other than C are still available on the market. The four mebendazole tablets currently available in South Africa were also tested and it was found that all of them contained polymorph C. PMID- 9876613 TI - Transdermal delivery of isosorbide 5-mononitrate from a new membrane reservoir and matrix-type patches. AB - Isosorbide 5-mononitrate (5-ISMN) has a direct relaxing effect on vascular smooth muscle. In the present study we developed matrix and reservoir-type transdermal patches of 5-ISMN. We investigated the usefulness of a new film-forming material isolated from the roots of Salacia macrosperma to serve as rate-controlling membrane for the reservoir-type patches. Matrix-type patches were formulated using polyvinyl chloride. Permeation studies through rat skin were conducted on both types of patches using Teflon cells. The mean +/- SD flux values from the matrix- and reservoir-type patches were 99.55 +/- 22.89 and 31.82 +/- 8.31 micrograms/cm2.hr, respectively. PMID- 9876614 TI - Effect of glycine/citric acid on the dissolution stability of hard gelatin capsules. AB - Gelatin capsule crosslinking is a well-known phenomenon that results in reduced dissolution of capsule products with the passage of time and/or under accelerated stability conditions. These studies describe one means of preventing capsule crosslinking by incorporating glycine and citric acid into a triamterene/hydrochlorothiazide 37.5/25 mg capsule formulation (triam/HCTZ). Triam/HCTZ without glycine and citric acid showed extensive capsule crosslinking and then failed the USP dissolution specification after a 4-week accelerated (40 degrees C/85% relative humidity [RH]) stability study. Triam/HCTZ containing glycine alone showed some improvement in the dissolution stability but did not prevent gelatin crosslinking. This formulation also failed dissolution specifications after a 4-week accelerated stability study. The same results were obtained when only citric acid was incorporated into the triam/HCTZ. However when glycine and citric acid were incorporated together into the triam/HCTZ, crosslinking was completely prevented. Dissolution profiles remained the same throughout 12-week accelerated stability studies, with little or no drop in the dissolution values throughout the test period. The above results were confirmed with follow-up studies using gemfibrozil and piroxicam as model drugs. Disintegration times for gemfibrozil and piroxicam capsule formulations without glycine and citric acid increased dramatically with observed pellicle formation, but there was little or no change in the disintegration time of the model drugs formulated with glycine and citric acid. The results of these studies demonstrated that when glycine and citric acid are present in some gelatin capsule formulations, pellicle formation or crosslinking of the capsule gelatin is prevented. PMID- 9876615 TI - Evaluation of fast disintegrants in terfenadine tablets containing a gas-evolving disintegrant. AB - Effects of four fast disintegrants on the dissolution of terfenadine tablets containing the gas-evolving disintegrant, CaCO3, were evaluated. In addition, effects of presence of starch along with the fast disintegrants on the dissolution of the tablets were examined. Dissolution data were treated to give dissolution parameters which reflected efficiency of the disintegrant combinations. The four fast disintegrants improved disintegration/dissolution of the original formulation. The relative efficiency of improvement was in the order crospovidone > Ac-Di-Sol > Primojel > low substituted hydroxypropylcellulose. The presence of starch advertently affected the role of the fast disintegrants. Scanning electron microscope studies revealed that starch covered the drug containing granules and other particles of the tablet. pH changes during dissolution of representative tablets in 0.1 N HCl solutions were determined at specific time intervals. The progressive decrease in rates of acid consumption as a function of the amount of starch, along with the SEM studies, suggested that a barrier existed around the tablet particles. The barrier was generated by the swelled starch grains and was responsible for the loss of the dissolution improving capacity of the fast disintegrants. Furthermore, the barrier interfered with the diffusion of the hydronium ions and therefore, impaired the function of the disintegrant combination. PMID- 9876616 TI - Peroral sustained-release film-coated pellets as a means to overcome physicochemical and biological drug-related problems. I. In vitro development and evaluation. AB - In vitro preformulation testing has shown that the solubility and dissolution rate of the model drug compound ucb 11056 are highly pH dependent. Considering this, different sustained-release (SR) oral dosage forms of ucb 11056 were developed aiming to obtain the most constant and complete release of the drug during transit in the gastrointestinal (GI) tract. Classical approaches based on the use of SR formulations such as hydrophilic matrix tablets or pellets coated with one film-forming polymer (Eudragit NE30D or L30D-55) did not fulfill all expectations on the basis of their in vitro evaluation, i.e., the drug release and pattern remained highly dependent on the pH of the dissolution medium. Therefore, taking advantage of the flexibility of release adjustment obtainable from coating of pellets with different kinds of pH-sensitive film layers, a quite satisfactory pH independence of the release characteristics was obtained using formulation blends of neutral and anionic acrylic polymers. For the selected SR pellets batch 15 coated with NE30D/L30D-55 (7:3), the tridimensional topographic representation of the drug release versus time and pH showed that, notwithstanding the pH-dependent aqueous solubility of the drug, the release profiles were relatively homogeneous for any pH value ranging between 1 and 7. PMID- 9876617 TI - Peroral sustained-release film-coated pellets as a means to overcome physicochemical and biological drug-related problems. II. Bioavailability and tolerance assessment in dogs. AB - Sustained-release (SR) dosage forms consisting of pellets coated with different pH-sensitive film layers, previously optimized in vitro with regard to pH independence of their drug release characteristics, were evaluated in vivo after single administration to Beagle dogs. In vivo performances were compared to those of a nonoptimized SR matrix tablet and a reference instant release (IR) capsule, in terms of the observed plasma pharmacokinetic profiles for the parent drug (ucb 11056) and its primary metabolite (ucb 26201), the bioavailability results, and the drug tolerance data. All SR dosage forms were seen to be effective in prolonging the relatively short biological half-life of the compound and in reducing the incidence of concentration-related side-effects, e.g., emesis, and of behavioral symptoms, e.g., restlessness, discomfort, and indisposition. The film-coated SR pellets offer a number of advantages over the monolithic SR matrix system in terms of a drug delivery pattern less dependent on pH changes in the gastrointestinal (GI) tract, a higher flexibility for adjusting and controlling the pharmacokinetic profiles, and a consequently more efficient approach for keeping all concentration-related side-effects under control. PMID- 9876618 TI - In vitro release of theophylline from poly(lactic acid) sustained-release pellets prepared by direct compression. AB - Poly(L-lactic acid), (L-PLA) pellets containing theophylline as a model drug were prepared with increasing bovine serum albumin (BSA) load of 10, 20, 30, 40, or 50% by direct compression. The drug release from pellets was studied in phosphate buffered saline (PBS, pH 7.4) at 37 degrees C. The annealing effect on theophylline release from pellets was also studied at 20, 30, 60, and 80 degrees C. In all cases, release kinetics followed the Higuchian mechanism with an initial burst effect followed by sustained release of theophylline during the experimental period. Increasing BSA load resulted in a linear increase in Higuchian release rates presumably because of the hydrophilic nature of BSA. Furthermore, BSA did not interact chemically with the polymer matrix and was held physically by the dense polymer matrix. However, drug release decreased with an increase in annealing temperature. Release of theophylline was higher from PLA BSA combination pellets compared to PLA pellets at temperatures below the glass transition temperature (Tg) of the polymer and lower for temperatures above Tg. The temperature effect on drug release may be attributed to both the reduction of core solubility in the bulk phase and the lowering of diffusibility of the polymeric membrane. No drug-polymer interactions or polymer degradation was observed within the experimental setup when studied by differential scanning calorimetry (DSC), infrared (FTIR) spectroscopy, and gravimetric methods. DSC studies of pellets showed no hints of microstructural changes (crystallinity) of the polymers. In our experiments, theophylline was released primarily by leaching through channels and not by polymer degradation. The release rate was dependent on BSA loading and annealing. It may be concluded that PLA pellets can be fabricated suitably using BSA and annealing to design sustained-release preparations of water-soluble drugs. PMID- 9876619 TI - Effect of physicochemical factors on the release kinetics of hydrophilic drugs from poly(L-lactic acid) (L-PLA) pellets. AB - Poly(L-lactic acid) (L-PLA) pellets intended for either parenteral or oral use were successfully prepared by a direct compression technique without the use of heat or organic solvents. Salicylic acid and theophylline were chosen as drug candidates. The drug release from pellets was affected by the compression pressure. The Higuchi plots of the drugs showed a t1/2 dependent drug release pattern. The release rates of these drugs from PLA pellets were directly correlated to their solubilities in the dissolution media. At lower pH (< 7), the release of salicylic acid was found to be slower than theophylline; however, at higher pH (> 7), the release of salicylic acid was faster than that of the theophylline. The release rate of salicylic acid was higher at higher pHs, which was related to the increase in solubilities. Pellets were annealed at 20, 40, and 80 degrees C. A lower release rate was observed with increasing temperatures. Above the glass transition temperature (Tg) of the polymer, the release of drugs was significantly decreased. The drug release was independent of the ionic strength of the media for both salicylic acid and theophylline. We showed earlier that no drug-polymer interactions or polymer degradation were observed when studied by differentials scanning calorimetry (DSC) and infrared spectroscopy (IR) (1). The release mechanism was primarily physical diffusion and leaching during the experimental period. We conclude that the release of low molecular weight (MW) drugs from the high MW L-PLA was independent of the pH and the ionic strength of the dissolution media, but was dependent on the polarity of the drug and formulation factors, such as compression pressure and annealing temperature. PMID- 9876621 TI - Statistical evaluation of in vitro dissolution of different brands of ciprofloxacin hydrochloride tablets and capsules. AB - The in vitro dissolution of ciprofloxacin from commercially available tablets and capsules in China was studied using the USP apparatus I to compare the product performance from nine different manufacturers. Cumulative release greater than 75% was obtained from all of the products tested within 45 min. However, statistically significant differences were found between some of the products when in vitro data were analyzed using the Weibull function, similarity factor (f2), and multivariate analysis of variances. PMID- 9876620 TI - Physical characteristics and release behavior of salbutamol sulfate beads prepared with different ionic polysaccharides. AB - Salbutamol sulfate beads were prepared using anionic and cationic polysaccharides, Gelrite and chitosan, respectively. Alginate beads were also prepared for comparison. The mean diameter, porosity, and drug content of the beads were determined. The beads were examined by scanning electron microscopy (SEM), DSC, and x-ray diffraction. The drug release from the beads was studied in 0.1 N HCl (pH 1.2), distilled water, and phosphate buffer (pH 7.4). The physical examination of the beads indicated the presence of drug crystals with no interaction between the drug and polymers. The drug release was dependent on the ionic properties of the polymers and the pH of the release media. In acidic pH, chitosan beads showed a rapid drug release, whereas a sustained drug release was obtained from Gelrite beads. In contrast, the drug release in phosphate buffer was rapid from Gelrite, and chitosan showed a sustained drug release. The results of drug release from Gelrite were comparable to that from alginate beads. Gelrite is recommended as an anionic polysaccharide for sustained-release preparations. PMID- 9876622 TI - Determination of betamethasone dipropionate and salicylic acid in pharmaceutical preparations by high-performance liquid chromatography. AB - The simultaneous determination of betamethasone dipropionate (BD) and salicylic acid (SA) in both ointment and topical solution was developed using high performance liquid chromatography (HPLC). The method was standardized using a LiChrospher 100 RP-18 (125 x 4 mm, 5 microns) column, acetonitrile tetrahydrofuran-acetic acid 1% (25:20:55 v/v), apparent pH 3.3, as mobile phase, and UV detection at 254 nm. The peak area response versus concentration was linear in a concentration range from 5.0 to 50.0 micrograms/ml of BD and from 20.0 to 200.0 micrograms/ml of SA. The correlation coefficients were 0.9997 for BD and 0.9987 for SA, and the relative standard errors of estimates were 1.38% for BD and 3.27% for SA. The coefficient of variation and the recovery average were, respectively, 0.41-1.15% and 100.09% for BD, and 0.57-0.95% and 99.79% for SA. PMID- 9876623 TI - Factors affecting zero-order release kinetics of porous gelatin capsules. AB - Porous gelatin capsules were prepared by entrapping gas bubbles or by drilling in the capsule wall with syringe needles. After hardening by formaldehyde, the capsules were supposed to provide a floating sustained release over 10 hr. Different pore sizes, pore numbers, pore orientational symmetry on the capsule wall, the amount of the effervescent material added to prepare the capsule, the exposure time to formaldehyde, and excipient selected were studied to evaluate their influences on the dissolution kinetics. The gelatin capsules were prepared by adding various amounts of effervescent material or by drilling in the capsule wall with a syringe and then hardening by formaldehyde. Verapamil was used as the model drug and starch or lactose was the only excipient. Verapamil and excipient were mixed and used to fill the prepared capsule. The release of verapamil from the capsules follows the zero-order drug release and was observed after the burst phase. The porous capsule, when filled with the mixture of active ingredients and excipient, can achieve a zero-order release system. PMID- 9876624 TI - pH-sensitive release of indomethacin using lactan-acetate microspheres. AB - Hydrophobic lactan acetate was prepared from hydrophilic lactan gum by chemical modification and it was used for a pH-sensitive drug delivery system. Lactan acetate microspheres were prepared by the solvent evaporation method. The size of the microspheres prepared from lactan acetate was below 5 microns. The drug loading efficiencies of microspheres were approximately 70 and 80% at the initial amount of drug 40 and 80 mg, respectively. The microsphere showed pH-sensitive swelling behavior. The swelling capacity of microspheres at pH 7.4 was much greater than that at pH 1.2. The pH of the medium significantly influenced the in vitro release rate; the rate at pH 7.2 was approximately 10 times faster than that at pH 1.2. A pH-sensitive drug release pattern was due to the escape of drug from the microsphere that disintegrated after swelling. PMID- 9876626 TI - Effect of polysorbates on drug release from wax matrices. AB - The purpose of this study was to investigate the effect of various types and amounts of polysorbates on potassium chloride release. Potassium chloride, which is a highly water-soluble model drug, was embedded into wax (containing surfactants) to produce a sustained-release dosage form. Various kinds of polysorbates were chosen as surfactants to control the dissolution profile. The release of the model drug was tested by rotating paddle method of USP 23 and the dissolution process was characterized by the Weibull distribution. The surface tension of the aqueous solutions of polysorbates was determined by a computer controlled Sigma 70 tensiometer. The application of polysorbates in more than 2% concentration did not alter either the release rate of the embedded potassium chloride, or the surface tension values of the aqueous solutions. The results of this study allow the determination of the optimal concentration of polysorbates in the case of the potassium chloride release. PMID- 9876627 TI - Preliminary evaluation of Leucaena leucocephala seed gum as a tablet binder. AB - The seed galactomannan of Leucaena leucocephala Lam. de Wit var.K-8 (family Leguminosae), a natural polysaccharide, with properties comparable to guar gum, was evaluated as a pharmaceutical binder. Characterization was done using studies of compressibility, micromeritic, and mechanical properties of granules prepared by wet granulation and subsequent studies on compacts, both containing 5% w/w of binder. The seed gum was subsequently used as a binder with a badly compressible material, paracetamol, and studied likewise. The seed gum compared will with standard pharmaceutical binders (starch and polyvinyl pyrrolidone [PVP] K30), at least for properties studied herein. PMID- 9876625 TI - Functional roles of phenylalanine(7) of thymic humoral factor-gamma 2 in the impaired blastogenic response of uremic T-lymphocytes. AB - The peptide analogs of thymic humoral factor-gamma 2 (THF-gamma 2) in which phenylalanine residue at the 7th position are replaced by phenylglycine (Phg), homophenylalanine (Hph), and 1-naphthylalanine (1-Nal) were synthesized by a solid-phase method and the immunological significance of the aromatic amino acid of this position was comparatively investigated. The in vitro restoring effect of the synthetic peptides on the impaired phytohemagglutinin (PHA) response of T lymphocytes from uremic patients was tested. The observed activities of these peptides were in order (1-Nal7) thymic humoral factor [THF]-gamma 2 > 4-Fluoro (Phe7) THF-gamma 2 > THF-gamma 2. However, the other two analogs, [Phg7] THF gamma 2 and [Hph7] THF-gamma 2, had no restoring effect even at a higher concentration. PMID- 9876628 TI - Enhancement of bioavailability of griseofulvin by its complexation with beta cyclodextrin. AB - Griseofulvin is a poorly soluble antifungal antibiotic drug, the solubility of which can be enhanced by complexation with beta-cyclodextrin. The inclusion complex was prepared by coprecipitation method in various molar ratios of 1:1, 2:1, 3:1, and 1:2 of the drug and beta-cyclodextrin, respectively. The inclusion complex was characterized and evaluated by UV-VIS spectral studies and FTIR. The in vitro drug release studies indicated that the 1:2 molar ratio complex form of the drug significantly increased the dissolution rate when compared to the free form. The acute toxicity studies clearly indicated that the beta-cyclodextrin complex was nontoxic and the safety range was close to other Griseofulvin formulations. The in vivo study of the beta-cyclodextrin was carried out in both animals and human beings by administering in four different rabbits and volunteers, respectively. Pellets made with Griseofulvin-beta-cyclodextrin complex also showed a significant increase in the dissolution of the drug, revealing that beta-cyclodextrin plays an important role in the solubilization of Griseofulvin. PMID- 9876629 TI - The economic and social consequences of work-related musculoskeletal disorders: the Connecticut Upper-Extremity Surveillance Project (CUSP). AB - A population-based telephone survey was conducted in Connecticut to determine the social and economic impact of work-related musculoskeletal disorders (WRMSDs). Only 10.6% of cases had filed for workers' compensation. Respondents had spent an average of $489 annually out-of-pocket. Only 21% of individuals who had had medical visits or procedures reported having them paid for by workers' compensation. The WRMSD cases reported much higher levels of difficulty in daily tasks rated by the activities of daily living (ADL) scale, with odds ratios (ORs) ranging from 8.2 (child care) to 35.2 (bathing). The cases were significantly more likely to have moved for financial reasons (OR = 2.41), including having lost a home (OR = 3.44). The cases were also significantly more likely to have lost a car due to finances (OR = 2.45), more likely to have been divorced (OR = 1.91), and less likely to have been promoted (OR = 0.45). The study supports significant externalization of costs for WRMSD out of the workers' compensation system and a substantial social and economic impact on workers. PMID- 9876630 TI - Human exposures to arsenic from consumption of well water in West Bengal, India. AB - The authors visited the State of West Bengal, India, in August 1996, as consultants to the World Health Organization (WHO). The general mandate of the mission was to formulate recommendations to the Government of India regarding its efforts to assist the Government of West Bengal in remedying health problems arising from the presence of arsenic in groundwater in excess of the WHO guideline limit of 0.05 mg/L in eight districts of the State. The authors held discussions with Government of India and Government of West Bengal officials, as well as scientists, engineers, and physicians studying the analytic, medical, engineering, and hydrogeologic facets of the problem. They conducted field visits to arsenic-affected villages; inspected health centers, including the laboratories conducting the analytic and clinical studies; and interviewed and examined local lay people, including many arsenic-poisoned patients. This overview of the arsenic contamination problem in West Bengal is based upon a review of the scientific literature and government reports and the authors direct, firsthand assessment. The authors hope that their recommendations will assist in the development of a comprehensive infrastructure and plan of action, which are necessary to control the epidemic of chronic arsenic poisoning now afflicting West Bengal. PMID- 9876631 TI - Health and safety in the Lesotho Highlands Dam and Tunnel Construction Program. AB - To review health and safety practices and to determine the occupational health status of dam workers in the Lesotho Highlands, a health and safety audit and a health study of 258 workers was undertaken. The workers were administered respiratory health questionnaires and underwent chest x-rays and lung function and audiometric testing. The audit revealed a low level of awareness of health and safety issues at all levels of management and a lack of occupational medicine and industrial hygiene systems. Noise-induced hearing loss occurred among 92% of the study subjects. Among the 158 dam construction workers, 5.4% had pneumoconiosis. With poorly developed laws and regulatory bodies, unsophisticated trade unions, and little client supervision, health and safety had a low priority in this construction project. Recommendations are made that are pertinent to health and safety management where international companies are operating in developing countries. PMID- 9876632 TI - A standard tool for the analysis of occupational lung cancer in epidemiologic studies. AB - A standard tool for the analysis of known and suspected causes of occupational lung cancer in population-based studies is proposed in order to allow comparable definitions of exposure or of categorizations of occupations. It is based on a list of occupations and industries known (list A) or suspected (list B) to be associated with lung cancer. The lists were translated into codes of the International Standard Classification of Occupations (1968) and the International Standard Industrial Classification (1971). The specificity of the categorization is compromised for some groups that are defined by highly specific production processes or exposures. Nevertheless, the grouping is based on a highly valid source of information in population-based studies. It is proposed for use in the assessment of the public health impact of occupational lung cancer. PMID- 9876633 TI - Occupational health in southern Africa--challenges to occupational epidemiology. PMID- 9876634 TI - Occupational epidemiology in agriculture: a case study in the southern African context. AB - Some challenges facing occupational epidemiology in developing countries are outlined in this case study of agriculture drawing on Southern African research. These include the characterization of exposures in resource- and data-poor environments typical of developing countries, the assessment of outcomes where cross-cultural and socio-environmental confounders may be substantial obstacles, and the impact of environmental exposures on workplace health. Traditional assignment of low priority to the chronic effects of low-dose exposures relative to acute morbidity in developing countries must be critically examined, as must the gender bias of much occupational epidemiology in agriculture. Advocacy issues involving child labor and the ethics of research among vulnerable groups deserve rigorous attention. It is argued that, if occupational epidemiology is to have meaningful impact on the health of the most marginalized groups of workers in developing countries, it must redefine itself in terms of a public health approach. The boundaries of epidemiologic inquiry need to be broad, and amenable to interfacing with policy research, using qualitative methods and participatory approaches. More so than in order industrial settings, epidemiologists must move from research to practice, seeking to take action where interventions are needed, and to evaluate such actions. PMID- 9876635 TI - Creating alliances for disease management in industrial settings: a case study of HIV/AIDS in workers in South African gold mines. AB - The epidemic of HIV/AIDS is at an advanced stage in many African countries, but little attention has been given to the impact that this will have in industrial settings. Using the Southern African mining industry as a case study, the authors consider the state of the HIV epidemic and discuss programs that have been undertaken to manage HIV. They critically analyze the reasons current interventions have had little impact on HIV among mine workers, tracing the lack of success to neglect of the social and community contexts within which HIV transmission takes place, as well as the lack of attention to the psychosocial processes and mechanisms underlying disease transmission. Finally, they present an intervention that aims to address the limitations of existing industrial programs and improve the management of sexually transmitted diseases (STDs), including HIV, in a particular occupational setting through creating alliances between a wide range of community stakeholders. The intervention aims not only to reduce STDs, promote awareness of HIV risks, and distribute condoms, as existing programs have done, but also to address the broader social, cultural, and community contexts that facilitate HIV transmission. PMID- 9876636 TI - Health impact of occupational risks in the informal sector in Zimbabwe. AB - Information about occupational health in the informal sector is lacking, despite its growing contribution to employment. The author describes a survey of occupational health in urban and rural informal-sector workers in Zimbabwe. Common hazards included poor work organization, poor hygiene, ergonomic hazards, hazardous hand tools, and chemical exposures, particularly to pesticides and solvents. An annual occupational mortality rate of 12.49/100,000 was half the formal-sector rate. Reported rates of 131 injuries/1,000 workers and 116 illnesses/1,000 workers exceeded formal-sector rates tenfold and a hundredfold, respectively, although the distribution of injuries by economic sector correlated significantly with formal-sector rates. The survey found high levels of musculoskeletal and respiratory illness, thought to be underdetected in formal systems. A fifth of the injuries had resulted in permanent disability, with little consequent job loss, but no compensation granted. The author recommends improvements to occupational health in the informal sector, and suggests a broader survey of occupational morbidity in all sectors of employment. PMID- 9876637 TI - Occupational health and safety policy in the UK: who is driving the agenda with the "greylisting" of health and safety activists? PMID- 9876638 TI - Globally asbestos-free automobiles. PMID- 9876639 TI - Tooth discoloration due to isocyanate exposure? PMID- 9876640 TI - The compartmentalization of Surgery. PMID- 9876641 TI - One thousand and six consecutive laparoscopic intraoperative cholangiograms. AB - Intraoperative cholangiography was successfully performed in 1,000 out of 1,006 attempts in 1019 consecutive cholecystectomies. There were 783 chronic, 95 acute, 61 fibrotic, 27 gangrenous and 40 cases of hydrops of the gallbladder in those laparoscopic cholecystectomies performed. unsuspected common duct stones were identified in 5% of the patients. There were no injuries resulting from intraoperative cholangiography performed via the cystic duct. In this large series, routine cholangiography was thought to be helpful in the prevention of common bile duct injuries and the establishment of abnormal anatomy. In non-acute cholecystitis, intraoperative cholangiography is necessary due to the importance of abnormal anatomy verification. The technique of laparoscopic cholecystectomy differs greatly from that of open technique, and, therefore, routine intraoperative cholangiography is strongly advised. PMID- 9876643 TI - Functional MR imaging of the porcine kidney: physiologic changes of prolonged pneumoperitoneum. AB - BACKGROUND: Increased intraabdominal pressure (IPA) during laparoscopy has been associated with decreased urine output. The purpose of this study was to use a noninvasive MRI technique to measure renal vessel flow velocity and change in differential renal medulla and cortex perfusion during pneumoperitoneum. STUDY DESIGN: Six female farm pigs underwent general endotracheal anesthesia and dynamic imaging following left ventricular (LV) injection of Gd-DTPA, utilizing a dual echo gradient echo sequence. MRI was repeated after three hours of continuous 15 mm Hg pneumoperitoneum in three study pigs and after three hours of monitored general anesthesia without pneumoperitoneum in three control pigs. Renal artery and renal vein flow velocities were calculated using cine phase contrast technique. Renal perfusion was independently measured by LV injection of radiolabelled microspheres. RESULTS: There was a decrease in mean renal vein flow velocity in the pneumoperitoneum group as compared to the control group. During pneumoperitoneum there was a similar percentage reduction in the perfusion of the cortex (-28%) and medulla (-31%); this corresponded with a decreased urine output. In addition, radiolabelled microspheres corroborated the similar decrease in both cortical and medullary perfusion rates during pneumoperitoneum. CONCLUSIONS: Prolonged IAP is associated with a decrease in renal vein flow velocity and urine output. There is a similar decrease in the renal medulla and cortex perfusion rates during pneumoperitoneum of 15 mm Hg. PMID- 9876644 TI - Laparoscopic appendectomy in children. AB - BACKGROUND AND OBJECTIVES: The advantages of laparoscopic appendectomy over open appendectomy have not yet been clearly demonstrated. The present study evaluated our early experience with laparoscopic appendectomy in children, in terms of its safety, effectiveness, technical difficulties, and economics. METHODS: We reviewed the records of 50 cases involving laparoscopic appendectomy performed at our affiliated institutions between September, 1994, and September, 1996. Patient age ranged from 6 to 18 years (mean, 14 years). Thirty-two patients had acute nonperforated appendicitis, six had perforated appendicitis, two had fibrosis of the appendix, and ten had a histologically normal appendix. RESULTS: In five patients the laparoscopic procedures were converted to open appendectomies because of technical difficulties. There were postoperative complications in four patients: one incomplete appendectomy which subsequently required an open appendectomy for completion, one pelvic abscess, one bowel obstruction, and one minor wound infection. CONCLUSIONS: Laparoscopic appendectomy is a safe and effective procedure. It takes longer operative time than open appendectomy. Length of hospitalization and incidence of postoperative complications are equivalent to those of open appendectomy. Economic benefits are difficult to assess at present. In sum, we believe that with better training, surgical techniques and equipment, laparoscopic appendectomy will eventually become the surgical procedure of choice in appendicitis. PMID- 9876645 TI - Video-assisted thoracic surgery: applications and outcome. AB - Thoracoscopy has been revived and expanded by recent improvements in endoscopic technology. The enhanced application and outcome of VATS (video assisted thoracic surgery) was retrospectively studied. Between 1992 and 1995, 82 patients underwent diagnostic thoracoscopy or interventional VATS. Indications included: shortness of breath with nonspecific x-ray abnormality (45%), pulmonary nodule (25%), pleural effusion/empyema (21%), pneumothorax (14%), and hemoptysis, chronic cough or lung consolidation (5%). Sixty-six (83%) of the procedures were completed thoracoscopically. Eight procedures (10%) required addition of a utility mini-thoracotomy and 6 procedures (7%) were converted to formal thoracotomy. Specific diagnostic and/or therapeutic applications of VATS included: inspection; lysis of adhesions; stapling of blebs; biopsy of lung, pleura, or mediastinal structures; drainage and decortication of empyema; mechanical and chemical pleurodesis; wedge resection; and segmental resection. Diagnosis was established and/or treatment completed in 95% of cases. Pathologic diagnoses included: interstitial pneumonitis (22%), cancer (19%), bullous disease (15%), cocci nodule (9%), and other (18%). There were twenty-two complications (28.9%) and four deaths (4.8%). All four deaths were from causes unrelated to the surgery. The most common complications were: residual pneumothorax or hydrothorax (7), failed pleurodesis (3), and prolonged incisional pain (2). The advantage of reduced chest wall and muscle trauma utilizing VATS as opposed to traditional thoracotomy translates to less patient discomfort. The excellent magnified visualization afforded by VATS offers the opportunity to successfully conduct diagnostic and therapeutic interventions in the chest with equal or better visibility. Our findings suggest that the applicability and success of VATS is greatly expanding and its complication rate is less or, at worst, comparable to traditional thoracotomy. PMID- 9876642 TI - Laparoscopy during pregnancy: a literature review. AB - OBJECTIVE: To review the literature regarding the role of laparoscopy during pregnancy, particularly adnexal mass and non-obstetric surgery, incorporating the results of a series of 9 cases of laparoscopy during pregnancy at our centers. MATERIALS AND METHODS: A Medline search was performed to review the literature, and the reference lists provided by those articles were further explored for citations regarding laparoscopic adnexal surgery, appendectomy, and cholecystectomy. Our series of 9 patients consisted of pregnant patients with adnexal mass or acute abdomen who would otherwise have undergone exploratory laparotomy. Follow-up data for these 9 cases were collected by office visits, inquiry to the primary referring physicians, and telephone calls to the patient. RESULTS: The literature search yielded 42 additional cases of operative pelvic laparoscopy and 51 cases of abdominal operative laparoscopy (cholecystectomy and appendectomy). The publications, particularly regarding cholecystectomy, were supportive of the laparoscopic approach during pregnancy. All of the patients in our series had favorable outcomes. CONCLUSIONS: Advanced operative laparoscopy has been successfully performed for certain indications during pregnancy. PMID- 9876648 TI - Office single puncture laparoscopy sterilization with local anesthesia. AB - BACKGROUND AND OBJECTIVES: Recently there has been a renewed interest in office based laparoscopy sterilization utilizing local anesthesia with conscious intravenous sedation. The safety of performing the procedure outside of a hospital environment has been questioned. The author attempts to determine the incidence of major complications with a review of the literature and a retrospective chart review of his personal cases. METHODS: During the period from 1971 to 1995, the author performed 1,753 laparoscopic procedures, utilizing local anesthesia in 1,562 cases and general anesthesia in the remainder. The focus of this chart study is the 1,190 laparoscopy sterilization cases performed in an office exam room setting, utilizing local anesthesia and conscious intravenous analgesia. Conventional 10 mm single puncture laparoscopy equipment was used. Major complications were defined as requiring laparotomy, blood transfusion, cardiopulmonary resuscitation, or emergency transfer to the hospital. RESULTS: The author's series contained one major complication, which was easily handled in the office environment. A review of the literature, including the author's series, revealed a major complication incidence of 5 out of 20,568 cases reviewed. None of these five complications would have required immediate laparotomy or blood transfusion to prevent a fatality. CONCLUSION: The author proposes that there is no documented evidence that office laparoscopy places the patient at increased risk for a life-threatening complication and should be considered by experienced gynecologic laparoscopists who are interested in cost containment. PMID- 9876647 TI - Comparison of outpatient laparoscopic cholecystectomy in a private nonteaching hospital versus a private teaching community hospital. AB - The development of laparoscopic cholecystectomy has allowed the introduction of outpatient surgery for biliary tract disease. However, there appears to be a wide variation of the interpretation of "outpatient surgery," ranging from discharge the same day to keeping patients for overnight observation. We prospectively reviewed the last 50 chole-cystectomies performed at Spartanburg Regional Medical Center, a private teaching institution, and Upstate Carolina Medical Center, a private nonteaching hospital. All cholecystectomies were performed by board certified surgeons or surgical residents under the supervision of board certified surgeons. Spartanburg Regional Medical Center's standard was 23-hour observation with 9 patients (18%) being discharged home the day of surgery. Upstate Carolina Medical Center's standard was discharge home (usually 4-8 hours after completion of the procedure) with 39 patients (78%) discharged the same day. No patient discharged the same day presented back with any significant complication. Comorbid disease, biliary pancreatitis, ascending cholangitis, gangrenous gallbladder, extreme age and living conditions and conversion to open were factors considered for admission. Intra-operative difficulty such as oozing, excessive adhesiolysis, postoperative nausea, vomiting or pain control were also indications for overnight admissions. The extra 15 to 19 hours for routine observation did not change any treatment for any of the 41 patients and resulted in additional cost to the hospital of approximately $15,000. We conclude that same day, outpatient laparoscopic cholecystectomy can be done safely with discharge home 4 to 8 hours postoperative without significant morbidity in selective patients. PMID- 9876646 TI - Laparoscopic staging of endometrial cancer: the learning experience. AB - OBJECTIVE: To evaluate our learning-curve experience with laparoscopic management of endometrial carcinoma. METHODS: Retrospective review of our first 125 patients with endometrial cancer who were managed laparoscopically. The patient population was reviewed in a chronological manner, noting patient demographics, operative procedure and times, estimated blood loss, hospital stay, complications, and pathology. RESULTS: Overall, the mean age was 68.6 years (range 29-89), the mean weight was 160 pounds (range 97-328), and the mean Quetelet index was 27.8 (range 17.8-56.4). Metastatic disease was discovered in 28.8% (17/59) of patients with grade 2 or 3 lesions. There was no statistically significant variation in any of these parameters throughout the study. Operative times for staging without lymphadenectomy decreased significantly from a mean of 163 minutes to 99 minutes (p < .001). Operative times for staging with lymphadenectomy decreased from a mean of 196 minutes to 128 minutes (p < 0.02). Hospital stay decreased from a mean of 3.2 days in the first quarter of our study to 1.8 days (p < .0001). The overall average complication rate of 4% (two enterotomies, two cystotomies, and a transected ureter) did not vary. However, the rate of conversion to laparotomy dropped significantly from 8% (2/25) to 0% (0/100). CONCLUSIONS: We found that operative times and hospital stays for laparoscopic staging of endometrial cancer continued to drop after 125 cases. While the ability to detect metastatic disease and the rate of major complications appear unrelated to length of the operator experience, the conversion rate to laparotomy decreases with operator experience. Learning-curve parameters must be recognized by physicians, patients, and researchers for a host of reasons. PMID- 9876649 TI - Gallbladder dyskinesia in children. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to describe clinico pathological characteristics in a group of children with motility disorders of the gallbladder and correlate the findings with cases receiving surgical treatment for gallstone during the same period. METHODS: Retrospective chart analysis of all cholecystectomies from January, 1990 to June, 1995. Analysis of demographics, symptoms and duration, associated illnesses, diagnostic studies, pathological stratification, length of stay, complications, follow-up and patient satisfaction. Statistical comparison of clinical variables between gallstone and dyskinesia patients was analyzed using chi-square, and analysis of variance (ANOVA). RESULTS: Twelve children (14%) of 85 underwent laparoscopic cholecystectomy during a 66-month period for gallbladder dyskinesia. Their mean age was 14 +/- 3 years (range 7 to 18). Ten patients were female and two were males for a 5:1 ratio. Classic biliary symptoms (RUQ abdominal pain and FFI) predominated for a mean of 48 weeks. A predisposing factor was previous family history of gallstones in five cases (42%). The diagnosis was obtained after gallbladder non-visualization in one child and low ejection fractions after CCK stimulated hepatobiliary scan studies in the remainder. Mean ejection fraction was 16.8%. Ten cases (83%) had mild to moderate chronic cholecystitis, and two children had unremarkable pathologic changes. These changes correlated with the mean duration of symptoms, not with ejection fraction volumes. After a mean follow-up of 17 months, 11 children are free of symptoms, and one continues with intermittent diarrhea. Comparison between calculous and dyskinesia patients showed that biliary dyskinesia children suffer more dyspepsia, undergo more diagnostic studies and have a significant family history of gallstones. CONCLUSIONS: Gallbladder dyskinesia (GD) is a motility disorder causing symptoms similar to those of gallstones, although the clinical picture is more protracted. Diagnosis is confirmed using CCK stimulated hepatobiliary scan ejection fractions after thorough diagnostic work-up for other gastrointestinal causes. Laparoscopic cholecystectomy is the treatment of choice. Most children present with mild to moderate changes of chronic cholecystitis depending on duration of symptoms. Clinical improvement is seen in most cases after surgery. PMID- 9876650 TI - Laparoscopic hysterectomy with automatic stapling devices. AB - PURPOSE: To evaluate outcomes including operating time, blood loss, length of stay (LOS), return to work and complications of laparoscopic hysterectomy performed with automatic stapling devices. METHODS: Between 6/11/91 and 11/23/95, 127 laparoscopic hysterectomies were performed with automatic stapling devices. On an average, 6 firings with the stapler were done per case. Postoperative telephone survey and retrospective review of records were done. RESULTS: Data averages for operating time, blood loss, LOS and return to work, respectively, were 90 minutes, 190 cc's, 1.1 day and 2 weeks. Significant complications included delayed postoperative bleeding in 4 patients, all of which occurred within the first 35 cases. One was controlled laparoscopically and 3 others required exploratory laparotomies. Since certain precautionary measures as described were taken, hemorrhagic complications were eliminated. CONCLUSIONS: Laparoscopic hysterectomy can be performed safely and effectively with automatic stapling devices in properly selected patients. A potential hazard inherent with this technique includes intraoperative and postoperative bleeding from the staple lines, the incidence of which can be minimized by taking certain precautionary measures such as the use of white cartridges only and bipolar desiccation of staple lines when indicated. PMID- 9876652 TI - Thoracoscopic assisted pneumonectomy. AB - A lung carcinoma with tumor involving more than one lobe or in close proximity to the mainstem bronchus often requires pneumonectomy for surgical cure. Inflammation, bulky tumors, and dense adhesions limit the operative field of vision and may result in the abandonment of procedures with potential for complete extirpation. This case illustrates the utility of thoracoscopy in visualization of the hilum and other neurovascular structures in a patient with a proximal tumor and dense intrathoracic adhesions. Successful resection was made possible by use of combined open and thoracoscopic modalities. PMID- 9876651 TI - Inferolateral retraction reduces the risk of thermal injury to biliary structures. AB - OBJECTIVES: Do various traction techniques significantly change the anatomic position during laparoscopic cholecystectomy? METHODS: 16 cadaveric liver specimens were dissected and measurements were taken between structures in the triangle of Calot. Measurements were taken while traction was placed on the infidubilum in the inferolateral, cephalad and anatomic positions. Thermal necrosis data was measured one week post-injury in a rat model for Non-contact Yag laser, Quartz, Sapphire tip, and electrocautery. RESULTS: Inferolateral traction provided statistically significant increases in distance (P < 0.01) between the critical biliary structures. The increase in length by cephalad traction on the gallbladder was not statistically significant. Depth of necrosis for the devices were: cautery 1.03 mm, sapphire tip 0.63 mm, non-contact Yag laser 2.13 mm, and bare quartz tip laser 1.05 mm. CONCLUSION: Inferolateral traction produced a statistically significant increase in distance between the critical biliary structures. This was not demonstrated with cephalad traction. We recommend avoiding thermal dissection in the Triangle of Calot due to the thermal devices necrosis depth in relation to the proximity of the biliary structures. PMID- 9876654 TI - Laparoscopy--the early attempts: spotlighting Georg Kelling and Hans Christian Jacobaeus. PMID- 9876655 TI - The case for ultrasound. PMID- 9876653 TI - Late complication of laparoscopic salpingoophorectomy: retained foreign body presenting as an acute abdomen. AB - BACKGROUND: Laparoscopy is widely used as a tool in many clinical situations allowing for diagnosis and/or surgical management in a minimally invasive fashion. Most laparoscopic cases are ambulatory and allow patients to recover quickly. Nonetheless, attention to surgical technique is paramount to avoid both short and long term complications. CASE: A 32-year-old woman had a laparoscopy and a reported left salpingoophorectomy for benign disease of the ovary in September, 1994. Shortly thereafter, in January, 1995, she was diagnosed with an intrauterine pregnancy and delivered in October of 1995 by spontaneous vaginal delivery. The pregnancy and delivery were both uncomplicated. The patient presented four weeks postpartum with clinical suspicion of appendicitis. However, at the time of laparotomy, the patient was found to have a retained foreign body from her prior laparoscopy in the right lower quadrant with a pelvic abscess and evidence of prior right salpingoophorectomy. The appendix appeared grossly normal. CONCLUSION: Laparoscopy is a safe, effective modality for various surgical and gynecologic conditions. Although laparoscopy is usually done on an outpatient basis, complications can manifest several weeks or months later. This case illustrates and reminds us of the importance of adherence to surgical laparoscopic principles. These include direct visualization when removing equipment and a complete count of surgical instrumentation to confirm the integrity of such at the end of each procedure. PMID- 9876656 TI - Endoscopic management of adnexal masses. AB - BACKGROUND: The laparoscopic management of suspicious adnexal masses and early ovarian malignancies is discussed with the aim of maintaining accepted oncologic treatment principles. Comparative survival data of patients with gynecological malignancies managed by laparoscopy or laparotomy are still very scarce and the survival of cancer patients must not be compromised by new techniques. It is time to closely analyze laparoscopy and determine if it has a positive impact on the diagnosis and treatment of ovarian malignancies. In this paper we will address the following points: 1) Which ovarian cysts can be surgically treated by laparoscopy (pelviscopy)? 2) Is staging laparoscopy an accepted technique? 3) Is laparoscopy, as a second-look procedure, of benefit? 4) Is laparoscopic staging, together with histologic tissue sampling, adequate surgical technique in inoperable ovarian cancer with ascites and peritoneal carcinomatosis? 5) Does endoscopic biopsy of ovarian cancer stage Ia change the destiny of a patient into ovarian cancer Ic? DATA BASE: The above questions are analyzed based on our experience with the laparoscopic treatment of 1,225 patients with ovarian cysts and 165 ovarian cancer patients stage I to IV treated immediately by laparotomy during the years 1992-1995. CONCLUSIONS: Ovarian cystic tumors with no signs of malignancy can be dealt with by laparoscopic means with the option of immediate conversion to laparotomy or within one week if an ovarian malignancy is diagnosed. Today sampling laparoscopic lymphadenectomy of both pelvic and para aortic is feasible and adequate. On a curative level, the number of lymph nodes to be resected has yet to be determined. The adnexa can be extracted from the abdominal cavity with bag extraction without the danger of spillage. The uterus can be removed transvaginally with laparoscopic assisted vaginal hysterectomy (LAVH). We must be cautious to advocate laparoscopy for ovarian cancer. However, it is an excellent tool when used as a staging procedure. A careful preoperative screening of the patient and an exact definition of existing cysts with imaging techniques allows us to frequently apply laparoscopic surgery for ovarian cysts, leaving only readily detectable cancer cases for laparotomy. Many gynecological oncologists employing staging and second-look procedures for ovarian cancer agree that initiating a case with laparoscopy may preclude laparotomy for many patients. Tumor propagation by performing a biopsy in FIGO stage Ia ovarian cancer patients does not occur if the patient receives adequate radical surgical treatment within one week. According to the reports of Sevelda et al. and Dembo et al., the degree of differentiation and the existence of ascites are more relevant to decreasing the five-year survival rate of patients with ovarian cancer stage I than the rupture of capsule or penetration of the tumor. A dependency on the first two parameters was found in these two large statistical studies. As the question of endoscopic operations for adnexal mass is predominantly put for the sanitation of small ovarian tumors (ovarian tumors with solid particles in the cysts can be put into the section of primary laparotomies) there remains a wide field of indications for the laparoscopic treatment of adnexal mass and ovarian cysts with benign indications. For many young patients with non-malignant ovarian lesions such as endometriosis, benign cysts, benign cystic proliferations and fibromas, a laparotomy can be avoided and these lesions treated by laparoscopy. PMID- 9876657 TI - Laparoscopic pyeloplasty in the animal model. AB - PURPOSE: Laparoscopic pyeloplasty has been associated with long operative times. This study proposed to evaluate the feasibility of two different laparoscopic techniques for the performance of pyeloplasty repair of secondary ureteropelvic junction (UPJ) obstruction. MATERIALS AND METHODS: Sixteen female Yucatan minipigs underwent general anesthesia for cystoscopy, retrograde pyelography, urine culture and a baseline renal scan. Unilateral UPJ obstruction was created by ligating the UPJ over a 5F catheter. Six weeks later a laparoscopic pyeloplasty was performed utilizing an intracorporeal suturing technique and the Lapra-Ty suture clip or the Endostitch device with intracorporeal knot typing. Four control animals underwent only cystoscopy and in/out ureteral catheterization. In the study animals the ureteral stent was maintained for six weeks and at six weeks, three months and six months post-pyeloplasty the animals underwent the previously mentioned studies. At six months post-pyeloplasty the animals were euthanized and the UPJ was calibrated. Histopathology was obtained on the ureter below the anastomosis, at the anastomosis, above the anastomosis and on a renal biopsy. RESULTS: All planned laparoscopic pyeloplasties were completed. However, the stricture model was too severe in that most animals developed 40-45% decrease in renal function in the kidney following ipsilateral UPJ ligation. There was no significant difference between the two pyeloplasty techniques with respect to operative time to perform the pyeloplasty (mean of 40 minutes), post-pyeloplasty ureteral caliber (7.5-8.0 F), serum creatinine or healing scores at, above or below the anastomosis. CONCLUSION: Laparoscopic pyeloplasty can be performed equally successfully with the Endostitch device and intracorporeal knot tying or with the intracorporeal suturing technique and Lapra Ty clips. The resultant pyeloplasty is also equivalent for the two techniques. PMID- 9876658 TI - Acute cholecystitis is an indication for laparoscopic cholecystectomy: a prospective study. AB - OBJECTIVE: Acute cholecystitis has been considered as a relative or absolute contraindication to laparoscopic cholecystectomy. The purpose of this study is to present our experience of laparoscopic cholecystectomy as a safe and effective treatment of acute cholecystitis. METHODS: Laparoscopic cholecystectomy was offered to 34 consecutive patients with acute calculous cholecystitis, diagnosed according to strict clinical and ultrasonographic criteria. We used only three trocars. The gallbladder was routinely aspirated and sharp graspers were used. We adopted the fundus-first method of dissection when safe identification of the Calot's triangle was difficult. The cystic duct was ligated whenever necessary. RESULTS: The procedure was completed in 31 patients. The mean length of the laparoscopic procedure was 43 minutes, their mean hospital stay was 2.8 days. For the open group the mean length of the operative procedure was 66 minutes, while the mean hospital stay was 5.3 days. The overall morbidity rate was low. CONCLUSIONS: The benefits of laparoscopic cholecystectomy can be safely extended to patients with acute cholecystitis. The operation must be done early in the course of the disease. The surgeon should have adequate laparoscopic experience and maintain a low threshold for conversion to open exploration. Modifications in technique should be adopted to achieve a successful outcome. PMID- 9876660 TI - Video-assisted thoracoscopy for the diagnosis of mediastinal masses in children. AB - BACKGROUND AND OBJECTIVES: Video-assisted thoracoscopy has been successfully used for several different thoracic procedures in adults. However, its use in children has been limited. The present study evaluated our experience with video-assisted thoracoscopy in the diagnosis of mediastinal masses in children. METHODS: Nine children (age range, 3 to 18 years) with undiagnosed mediastinal masses underwent video-assisted thoracoscopy. The operation was performed using general anesthesia, with the patient intubated with a single lumen endotracheal tube. RESULTS: In all cases adequate tissue for diagnosis was obtained. Three patients had tuberculosis, two had Hodgkin's disease, two had granuloma, one had metastatic Wilms' tumor, and one had thymic hyperplasia. There were no complications related to the operative procedure. CONCLUSIONS: Video-assisted thoracoscopy for the diagnosis of mediastinal masses in children is a safe procedure. It provides good visualization, access to the mediastinum and adequate tissue for diagnosis. Further, postoperative discomfort is tolerable and cosmetic results are excellent. PMID- 9876659 TI - Comparison of office hysteroscopy, transvaginal ultrasonography and endometrial biopsy in evaluation of abnormal uterine bleeding. AB - OBJECTIVE: A comparison between office hysteroscopy, transvaginal ultrasonography and endometrial biopsy was performed, in terms of detection of intrauterine lesions. A secondary objective was assessment of evaluatory approach in the management of abnormal uterine bleeding in an outpatient setting. DESIGN: Prospective observational study. MATERIAL AND METHODS: A total of 54 women were evaluated for abnormal uterine bleeding. Assessment included performance of an endometrial biopsy, a transvaginal ultrasound scan followed by office hysteroscopy. Results of hysteroscopy were taken as the gold standard. Sensitivity and specificity of the investigations were assessed. The bleeding pattern was classified as heavy regular, irregular, postmenopausal and heavy or unscheduled bleeding on hormone replacement therapy. RESULTS: The incidence of focal intrauterine lesions in patients presenting with abnormal bleeding was 52% for all ages and 31% for the postmenopausal group. Seventy-five percent of the patients with Hb < 11 gm% and 67% with an enlarged uterus harbored a focal pathology. The incidence of lesions in patients with heavy regular bleeding was 74%. The sensitivity and specificity of transvaginal ultrasound when compared with results of hysteroscopy was 0.60 and 0.88 respectively. A normal endometrial biopsy had a negative predictive value of 51%. The sensitivity and specificity of endometrial biopsy were 0.04 and 0.83, respectively. CONCLUSION: Both transvaginal ultrasound and endometrial biopsy exhibited poor sensitivity for detection of focal intrauterine lesions. Considering the significantly high incidence of intrauterine lesions in patients presenting with abnormal bleeding, the most cost-effective approach appears to be proceeding with hysteroscopy early in assessment. PMID- 9876661 TI - Subfascial endoscopic perforator surgery: new life for an old procedure? AB - BACKGROUND: Division of incompetent perforating veins has long been regarded as an appropriate approach for treatment of venous stasis ulcers. The development of endoscopic techniques using standard laparoscopic instrumentation has permitted the application of this therapy without the need for long open incisions, fraught with complications. METHODS: We report our experience with 20 cases of subfascial endoscopic perforator surgery (SEPS) in 19 patients. Seventeen limbs had active ulceration at the time of operation. A gas insufflation technique with two 10 mm ports was used in most cases. RESULTS: An average of four perforating veins were divided in each case. Mean operating time was 1.5 hours. At a mean follow-up of eight months, initial complete healing occurred in 14 of 17 ulcers, three ulcers improved, and three healed ulcers at the time of SEPS have remained healed. One patient developed a small area of recurrent ulceration after initial healing. There were no thromboembolic complications. One procedure was technically unsuccessful because of morbid obesity. One patient developed a wound infection, and one patient required re-exploration for a subfascial hematoma. CONCLUSION: SEPS is a safe, minimally invasive procedure which should become an important part of the surgical armamentarium in treating patients with venous ulcers. PMID- 9876662 TI - A comparison of epidural versus general anesthesia for outpatient endoscopic preperitoneal herniorrhaphy. AB - OBJECTIVES: To study the efficacy of epidural versus general anesthesia on length of stay, patient recovery and anesthetic-related complications in patients undergoing endoscopic preperitoneal herniorrhaphy. METHODS: One hundred sixty seven consecutive patients undergoing endoscopic preperitoneal herniorrhaphy from July, 1994, to August, 1995, were retrospectively studied. A total of 243 herniorrhaphies were performed. Four patients required conversion of epidural anesthesia to general anesthesia because of inadequate sensory blockade (67/71; 94% success rate). One-hundred-forty-eight patients were available for review. Sixty-seven patients underwent successful epidural anesthesia during the case, while 81 patients were managed with general anesthesia. RESULTS: Thirty patients (37%) receiving general anesthesia required interventions for nausea compared to only six patients (9.0%) in the epidural anesthesia group (p < 0.001). Thirty patients (37%) in the general anesthesia group required intervention because of complaints of pain, compared to 13 (19.4%) in the epidural group (p < 0.05). There were no differences between the two groups for length of stay in OR, PACU, or total hospital times. CONCLUSIONS: The use of epidural anesthesia during the performance of endoscopic preperitoneal herniorrhaphy was associated with a decrease in the incidence of postoperative pain and nausea. The technique was successful in 94% of the cases in which it was used. Epidural anesthesia is recommended as an effective alternative to general anesthesia for the performance of outpatient endoscopic preperitoneal herniorrhaphy. PMID- 9876663 TI - Abscess formation following spilled gallstones during laparoscopic cholecystectomy. AB - OBJECTIVE: Our purpose was to report the occurrence of abscess following spilled gallstones during laparoscopic cholecystectomy as experienced at Good Samaritan Hospital, and to compare it to the experience of other institutions in order to identify the incidence, characteristics and possible risk factors for the development of this complication. METHODS: Four case reports of abscess following spilled gallstones during laparoscopic cholecystectomy are presented. The English literature was reviewed, and characteristics of the case reports found in the literature were compared. RESULTS: In four years at this institution, four reports of abscess formation following laparoscopic cholecystectomy have been identified. Two occurred in elderly females and were located in the right flank in both. Two were in middle-aged men, both diabetic. One abscess was in the right flank and one in the right pleural space. Twenty-three cases were found in the literature. The average age was 65 years; there were 12 men and 11 women. The locations of abscess formation were trocar sites (most common), right subphrenic space, right flank or retroperitoneum, and pelvis. The average time to presentation was 4-5 months (range 4 days-12 months). CONCLUSIONS: Abscess formation following spilled gallstones during laparoscopic cholecystectomy occurs infrequently, but can be debilitating and require more than one procedure. The older population appears to be at greater risk. Future prospective studies of patients who undergo laparoscopic cholecystectomy are needed to more accurately determine the incidence of this complication and to identify the population at risk. PMID- 9876664 TI - Spontaneous erosion of a lost intra-abdominal gallstone through the back eight months following laparoscopic cholecystectomy. AB - BACKGROUND AND OBJECTIVES: Gallbladder perforation during laparoscopic cholecystectomy with spillage of bile and gallstones occurs in up to 40% of patients. Several reports have recently been published describing complications related to these lost gallstones. The purpose of this study was to determine the incidence of this complication in our patients. METHODS: A prospectively maintained database of 856 laparoscopic cholecystectomies performed between 1989 and 1996 by a single surgeon was analyzed. RESULTS: The number of perforations resulting in loss of stones in the abdominal cavity was 16% (165 patients). Of these 165 patients, only a single patient could be identified as having a long term complication. CONCLUSIONS: Intra-abdominal lost gallstones can produce complications secondary to migration and erosion. It is prudent to make a concerted effort to remove spilled gallstones by every possible means but conversion to laparotomy is not justifiable. PMID- 9876665 TI - Vascular complications following laparoscopy: two unusual cases. AB - BACKGROUND: Vascular complications following laparoscopic techniques may often be attributed to the incomplete control of bleeding sites at laparoscopy. When confronted with post-laparoscopy symptoms of hemodynamic insufficiency, the surgeon may infer the existence of hemorrhagic complications neglected at the laparoscopic session. METHODS: The author reviewed two otherwise normal cases of laparoscopic procedures that were complicated by bleeding disorders of unknown origin. RESULTS: Diagnosis and treatment of the hematologic complications revealed causes other than operator-inflicted injury. CONCLUSIONS: While laparoscopists should remain vigilant concerning the very real threat of overlooked vascular injury following laparoscopy, some patients may exhibit hemorrhagic symptoms unrelated to the laparoscopic procedure. PMID- 9876666 TI - Laparoscopic pyelolithotomy in a pelvic kidney: a case report and review of the literature. AB - BACKGROUND AND OBJECTIVES: Laparoscopic pyelolithotomy was performed in a pelvic kidney with a large renal pelvis calculus. METHODS AND RESULTS: Laparoscopic pyelolithotomy was successfully performed in a pelvic kidney with an operative time of 310 minutes. The use of intraoperative fluoroscopy and a semi-automatic suturing device greatly facilitated the procedure. The patient's operative pain was managed with 3 doses of ketorolac; she resumed a regular diet the day after surgery, and was discharged on the first postoperative day. CONCLUSIONS: For patients with a large stone in the renal pelvis of an ectopic kidney, laparoscopic pyelolithotomy provides an effective approach. PMID- 9876667 TI - Gallbladder volvulus with gangrene. Case report and review of the literature. AB - BACKGROUND: Volvulus of the gallbladder is an unusual event. Over 300 cases have appeared in the literature since first reported by Wendel in 1898, ranging in age from 2 to 100 years old. Unusual mobility of the gallbladder due to congenital anomalies is a constant factor in all occurrences. CASE REPORT: This is a report of a laparoscopic cholecystectomy in an 82-year-old white female with volvulus of the gallbladder presenting with a chief complaint of chest pain. CONCLUSION: The patient had an uneventful recovery and was discharged within 72 hours after surgery, indicating that with the proper technique laparoscopic cholecystectomy is both feasible and safe in gallbladder volvulus. PMID- 9876668 TI - Suture? Staple? Electrosurgery? How to decide what is best for you. PMID- 9876669 TI - Using delta/DRG diagrams and decision tree analysis to select a cost-effective surgery for cholecystitis. AB - BACKGROUND AND OBJECTIVES: Many studies have attempted cost analysis of laparoscopic cholecystectomy as compared to open cholecystectomy. However, these analyses have included costs, charges, expenses, etc., and at times they have been used interchangeably. This paper demonstrates how DRG diagrams containing charges and length-of-stay, preoperative prediction of conversion rates, decision tree construction and sensitivity analysis can be used to select the most cost efficient operation for a given patient with cholecystitis. METHODS: A Delta DRG analysis for complicated cholecystectomy (DRG 195) showed the hospital to be an extreme outlier in both charges and length of stay. Record review indicated that 55% of the cases were converted laparoscopic cholecystectomies and the remainder were aged or younger patients with advanced disease. Chart and literature review determined the causes and the probability of conversion. Data were then placed into decision-tree and sensitivity analyses. The most cost-effective operation for a given probability of conversion was demonstrated. RESULTS: Three preoperative findings and combinations of each predicted conversion rates and analysis showed that the charge of laparoscopic cholecystectomy must be held below the range of $5,361-$13,084 to make routine laparoscopic cholecystectomy cost-effective. CONCLUSIONS: This method demonstrated that using Delta/DRG, decision-tree and sensitivity analysis offers physicians, hospitals and other health-care providers a method of evaluating the treatment of DRG categories to determine the most cost-effective management. PMID- 9876670 TI - "AEIOU: the ABC's" of conversion from laparoscopic to open cholecystectomy. AB - BACKGROUND AND OBJECTIVES: To examine and classify the reasons for conversion and the points at which laparoscopic cholecystectomies are converted to open procedures and whether these change over time. METHODS: This is a retrospective study of all patients undergoing cholecystectomy from June 1, 1990 to June 30, 1995. Reasons for conversion were classified using the "AEIOU:ABC" system developed for this study and conversion points were assigned chronologically. RESULTS: The "AEIOU:ABC" classification system was utilized. The most common reasons for conversion were: acute inflammation N = 61 (26.1%); adhesions N = 51 (21.8%); and organ system pathology N = 39 (16.7%). The most common conversion points were; after visualization of the peritoneal cavity but prior to dissection of the cystic structures N = 103 (44.0%); dissection of the cystic structures N = 58 (24.8%); initial laparoscopy N = 36 (15.4%). When the reasons for conversion were evaluated for changes over time there was no statistically significant change for the total group or any individual surgeon. Conversion points did not change with increasing operative experience. CONCLUSION: The "AEIOU:ABC" classification system is a simple, effective and easy to use system for classifying the myriad of reasons for conversion. The system needs to be validated prospectively not only for laparoscopic cholecystectomy but for possible application to other laparoscopic procedures. PMID- 9876671 TI - Laparoscopy between the world wars: the barriers to trans-atlantic exchange. Spotlighting Heinz Kalk and John C. Ruddock. PMID- 9876672 TI - Lasers in surgery: a return to reason. PMID- 9876673 TI - Operative laparoscopy: redefining the limits. AB - The continually changing definition of operative laparoscopy as well as the ever widening boundaries of its use are discussed in this report. It is important to prepare residents to adequately undertake advanced laparoscopic surgery as laparotomy is gradually replaced by laparoscopy for many routine procedures. Since degree of training and experience strongly correlate with complication rates, more focus on laparoscopy during graduate education would be beneficial to residents in order to keep them up to date with the rapid development of this field. PMID- 9876675 TI - Intraoperative laparoscopy by a flexible scope: is it reliable in contralateral childhood hernias? AB - BACKGROUND AND OBJECTIVES: Negative contralateral groin exploration for childhood hernias raises the question of whether contralateral groin exploration is necessary or not. To find out whether a contralateral processus vaginalis was patent, we performed laparoscopy with a flexible scope. METHODS: After carbon dioxide insufflation, a flexible laparoscope was inserted through the opened hernia sac and the contralateral processus vaginalis orifice was examined. We considered a patent processus vaginalis as a potential hernia. The study involved 20 children: 16 boys and 4 girls. The symptomatic side was explored in a conventional manner and laparoscopy was performed through the opened hernia sac. RESULTS: A contralateral processus vaginalis was found in 6 children: 4 boys and 2 girls. These results were confirmed by exploring the opposite groin. We did not explore if the laparoscopic examination was within normal limits. There was one false-positive result in a female patient. CONCLUSIONS: Intraoperative non puncture laparoscopy utilizing a flexible laparoscope through the hernia opening is an uncomplicated, reliable and precise method for identifying a patent contralateral processus vaginalis. It may represent a satisfactory alternative to routine bilateral inguinal exploration. Also, use of the flexible laparoscope may be more beneficial than use of a rigid laparoscope passed through the umbilicus or hernia sac. PMID- 9876674 TI - Laparoscopic colectomy: the absolute need for a standard operative technique. AB - BACKGROUND: The aim of this study is to review our experience performing laparoscopic colon surgery and to present the operative technique as used and standardized by us. METHODS: From April 1992 to December 1996, 158 consecutive patients underwent laparoscopic colon surgery. There were 92 females and 66 males, whose average age was 66.7 years (range 31-92); 134 patients (84.9%) were operated on for carcinoma, and the remaining 24 (14.1%) or benign disease. RESULTS: There were 117 procedures completed laparoscopically out of 158 patients (74%); 103 colon resections (18 for benign disease and 95 for malignant disease), 7 Hartmann procedures, 3 for reversal of Hartmann's procedures, 1 rectopexy, and 3 ileotrasversostomies. Conversions were required in 41 out of 158 cases (25.9%); 19 of these cases, however, were converted to a laparoscopic-facilitated procedure. The most common causes for conversion were the presence of bulky tumors and/or tumors that contaminated adjacent structures (16/158), adhesions due to previous operations (8/158) or patient obesity (5/158). There were 31 complications (19.6%), 9 of which required re-operation. There was only one recurrence (0.9%) that manifested 15 months after the procedure, at both trocar and drainage sites, and with peritoneal carcinomatosis. This occurred in a patient with rectal neoplasia who suffered a perforation of the rectum during dissection, with bowel spillage. The average number of lymph nodes harvested in resected specimens was 12.8 (range 1-41), whereas the mean distance of the tumor from the proximal margin of resection was 11.5 cm (range 5-35), and from the distal margin 7.5 cm (range 1-25). The average operative time was 165 minutes (range 40-360), and the mean hospital stay was 9.2 days (range 6-40). There were three mortalities out of 158 patients (1.9%). CONCLUSIONS: Laparoscopic colon resection for malignant lesions, performed with the highest respect for oncologic principles, has demonstrated that it is difficult to develop a barrier to wall and intraluminal recurrence. Recurrence, in our opinion, is caused by improper surgical technique. Therefore, neoplastic colon laparoscopic surgery must be the prerogative of selected and specialized centers. PMID- 9876676 TI - No scalpel, no iv, no stitch, microendoscopic office laparoscopy under local anesthesia. AB - BACKGROUND: The medical literature is replete with articles verifying the usefulness of laparoscopic procedures under local anesthesia. Recent research has examined the efficacy of microendoscopy with local anesthesia. In this series of patients, we focused on new technology to determine if microendoscopy could be utilized in an office setting. METHODS: Between July 1994 and April 1995, we performed 51 microendoscopic office laparoscopy under local anesthesia (MICRO OLULA) using the 1.5 mm Pixie laparoscope by Origin, a 1.7 mm laparoscope by Optimed and 5 mm laparoscope by Jarit. All cases were performed in an office operating room at the Women's Medical Plaza in Montgomery, Alabama. Only one patient was unable to have the procedure completed due to intolerance under local anesthesia. RESULTS: Fifty-one micro-olulas were performed on these patients who had an average age of 31 years and an average weight of 157 pounds. Intraoperative abdominal time averaged 3 minutes. One case was done with a 5 mm laparoscope and five cases with the 1.7 mm Optimed laparoscope. The 1.5 mm Pixie laparoscope was used in 45 patients. CONCLUSIONS: Our patients seemed to like the idea of a small device to view their pelvic cavities. The small laparoscopes provide excellent cosmesis, and laparoscopes deserve further development and clinical trial to determine their most advantageous use in the office setting. PMID- 9876677 TI - A comparison of patient recovery following unilateral and bilateral endoscopic preperitoneal herniorrhaphy. AB - INTRODUCTION: The advantage of using minimally invasive techniques over open techniques in the repair of inguinal hernias remains unclear. One of the more established indications for the performance of minimally invasive (e.g. endoscopic preperitoneal) herniorrhaphy is the presence of bilateral hernias. However, no prior study has compared the recovery following unilateral and bilateral endoscopic preperitoneal hernia repairs. PATIENTS AND METHODS: From July 15, 1994 through August 16, 1996 one primary surgeon performed 373 hernia repairs on 250 patients. Unilateral herniorrhaphy (UH) was performed on 114 males and 13 females with an average age of 58 (range 18-89). Bilateral herniorrhaphy (BH) was performed on 121 males and 2 females with an average age of 53 (range 18 86) (p > 0.05). Within the UH group there were 105 virgin hernias and 22 recurrent hernias. The BH group included 212 virgin hernias and 34 recurrent (p > 0.05). Bilateral repairs took longer to perform than unilateral repairs (65 minutes vs. 45 minutes) (p < 0.05). At the time of discharge, all patients were given a postoperative survey and asked to record their level of pain, narcotic use and level of activity on the day of surgery and postoperative days 1, 2, 3, 7, 14, and 28. RESULTS: No differences were found in pain perception, narcotic use or level of activity on any of the days measured between the two groups (p > 0.05). In addition, both groups returned to work at a similar time (UH: 6.32 +/- 3.29 days, BH: 6.68 +/- 4.13 days) (p > 0.05). CONCLUSION: Bilateral endoscopic preperitoneal herniorrhaphy can be performed with the same expected patient recovery as unilateral repairs. PMID- 9876678 TI - Mirizzi syndrome type II: is laparoscopic cholecystectomy justified? AB - Mirizzi syndrome type II is an uncommon cause of obstructive jaundice caused by an inflammatory response to an impacted gallstone in Hartmann's pouch or the cystic duct with a resultant cholecystocholedochal fistula. Two cases of Mirizzi syndrome type II are presented. Clinically only one patient had jaundice and endoscopic retrograde cholangiopancreatogram (ERCP) established a preoperative diagnosis of Mirizzi syndrome. The other patient's diagnosis of Mirizzi syndrome was made intraoperatively. It is important to properly identify the anatomy at the time of surgery to avoid compromising the common bile duct. Operative treatment of Mirizzi syndrome type II includes laparoscopic or open subtotal cholecystectomy; placement of a T-tube with either laparoscopic or open cholecystectomy; or creation of a hepaticojejunostomy with cholecystectomy. Although there is a report of laparoscopic treatment of this syndrome without long term follow-up, we believe that once there is any question of injury to the common bile duct, safety demands that the laparoscopic procedure be converted to an open one with implementation of appropriate therapy. PMID- 9876680 TI - Polypoid lesions of the gallbladder in children. AB - Polypoid lesions of the gallbladder in children are rare. We report a case of a gallbladder polyp in a 14-year-old boy who presented with recurrent right upper quadrant abdominal pain. Ultrasound examination of the abdomen revealed a polypoid lesion of the gallbladder. His symptoms resolved after laparoscopic cholecystectomy. Histological examination of the gallbladder demonstrated a benign adenomatous polyp. Although the experience with polypoid lesions of the gallbladder in children is limited, we currently recommend cholecystectomy because these lesions are associated with acalculous cholecystitis, and because their long-term effects are unknown. PMID- 9876679 TI - Laparoscopic management of gallbladder disease in children and adolescents. AB - BACKGROUND: Between July 1991 and April 1996, 40 children and adolescents age 17 or less underwent laparoscopic management of their gallbladder disease. Twenty eight patients were females and 12 were males. Their average age was 12.7 years (range 2-17 years), and average weight was 50 kilograms, range 12.2-95.9 kilograms. Nine patients had gallstone pancreatitis and seven patients had sickle cell disease. Laparoscopic cholecystectomy was attempted in all patients with or without modifications of the standard technique dictated by the size of the patient. METHODS: The practice of intraoperative cholangiogram varied with the practicing surgeon. Those with clinical or biochemical evidence of common bile duct obstruction underwent preoperative endoscopic retrograde cholangiopancreatography to rule out other causes of hyperbilirubinemia and/or therapy for choledocholithiasis if present. Patients with unsuccessful intraoperative cholangiogram were followed clinically and were subjected to postoperative endoscopic retrograde cholangiopancreatography should they develop clinical or biochemical evidence of common bile duct obstruction. Thirty-six patients were completed laparoscopically (90%). Four patients were converted to open cholecystectomy (10%). Four patients required preoperative endoscopic retrograde cholangiopancreatography and were successfully treated. Postoperative endoscopic retrograde cholangiopancreatography was unsuccessful in one patient who required the procedure because of retained common bile duct stone. Four patients suffered complications (10%). Three patients continued to have abdominal pain that was not helped with surgery. CONCLUSIONS: Based on our experience, laparoscopic cholecystectomy with preoperative endoscopic retrograde cholangiopancreatography if required, is safe and effective in management of gallbladder disease in children and adolescents. However, careful preoperative evaluation is required to avoid persistent postoperative abdominal pain. PMID- 9876681 TI - Video-assisted thoracoscopic surgery in the management of pediatric empyema. AB - BACKGROUND: Management of empyema in children has progressed from open thoracotomy to video-assisted thoracoscopic surgery (VATS). The purpose of the present study was to evaluate the efficacy and safety of VATS in children with multiloculated empyemas. METHODS: Nine children (mean age, 4 years; range, 21 months to 13 years) with empyema, in whom multiple loculations were found on computed tomography, were treated with VATS from January, 1994, to November, 1996. All patients underwent VATS under general anesthesia, with drainage of the empyemas, decortication, and placement of chest tubes under direct vision. RESULTS: In all nine patients, VATS was successful. Average operating time was 120 minutes. Blood loss was insignificant, except in one patient who needed an intraoperative blood transfusion. This child required extensive decortication, with blood oozing from raw areas. All patients recovered well, with no recurrences to date. An algorithm for the use of VATS in the treatment plan for children with empyema was established. CONCLUSION: VATS provides safe and effective treatment in the management of pediatric empyema. Moreover, it avoids lengthy hospitalization, prolonged intravenous antibiotic therapy, and unnecessary pain and stress secondary to placement of chest tubes without anesthesia. PMID- 9876682 TI - Video-assisted thoracoscopic management of post-pneumonectomy empyema. AB - BACKGROUND: Post-pneumonectomy empyema is a major therapeutic challenge in thoracic surgery. The presence or absence of a concomitant bronchopleural fistula directs treatment of this condition. When there is no bronchopleural fistula the condition is classically treated with thoracostomy drainage, irrigation and antibiotic instillation with closure. This approach is, however, associated with a significant rate of primary failure. Alternative modified techniques involve opening the thoracic cavity widely with serial debridement followed by interval closure. Multiple surgical procedures often require a protracted hospital stay. METHODS: We describe a technique in three patients utilizing video-assisted thoracoscopic surgery for debridement and closure of the pneumonectomy cavity. CONCLUSION: Advantages of this technique include debridement under direct visualization, low morbidity, and potential for a shorter hospital stay. PMID- 9876683 TI - Alternatives to total abdominal hysterectomy. PMID- 9876684 TI - The cystic vein: the significance of a forgotten anatomic landmark. AB - The cystic vein, a portion of biliary anatomy whose insignificance in open gallbladder surgery led to its being relegated to mythology, has been rediscovered by the magnified view of laparoscopic surgery. Its presence is an important anatomic feature that helps distinguish between cystic duct and common hepatic duct, thus diminishing the risk of inadvertent bile duct injury during laparoscopic cholecystectomy. PMID- 9876685 TI - Pelvic gliomatosis within foci of endometriosis. AB - The third reported case of pelvic gliomatosis found within foci of endometriosis is documented 16 years after the removal of a benign cystic teratoma. Grossly at laparoscopy the lesions appear as typical deep fibrotic endometriotic implants. PMID- 9876687 TI - Trocar site closure: a simple, inexpensive technique. AB - Trocar site complications continue to plague laparoscopic surgeons. Described in this report is an inexpensive simple technique to close all layers of the trocar site defect. PMID- 9876686 TI - The suprapubic cruciate incision for laparoscopic-assisted microceliotomy. AB - OBJECTIVES: To introduce cruciate microceliotomy for performing conventional open surgery maneuvers at laparoscopy without conversion to standard laparotomy. METHODS: Retrospective review of all pelvic operations performed by this method by a single surgical team at one institution from 1993 through 1996. RESULTS: We found the cruciate incision to provide excellent exposure for the performance of extensive uterine suture-reconstruction, morcellation, tubal microsurgery, and bowel surgery with excellent cosmetic and convalescent results in a series of 94 patients. No wound-related complications were noted. Postoperative analgesic requirements averaged 4.1 oxycodone/acetaminophen tablets per day, and hospital stay averaged 36 hours. CONCLUSION: In our experience, cruciate microceliotomy represents a useful, minimally-invasive adjunct for the performance of open surgical maneuvers at laparoscopy. PMID- 9876688 TI - Laparoscopic repair of abdominal hernias using an ePTFE patch--a modification of a previously described technique. AB - BACKGROUND: A variety of laparoscopic procedures for the repair of abdominal wall hernias have been described. The repair described in this paper represents a modification and improvement of one approach. METHODS: Our technique employs an Origin Tacker (Origin Medsystems, Inc, Menlo Park, CA) and a Gore suture passer (W. L. Gore, Flagstaff, AZ) to secure an expanded polytetrafluoroethylene (ePTFE) patch, (DualMesh, W. L. Gore, Flagstaff, AZ) to the anterior abdominal wall. RESULTS: This approach simplifies the repair by minimizing the number of steps required to secure the ePTFE graft to the anterior abdominal. CONCLUSIONS: We have found this to be a safe and reliable technique that may be used in the laparoscopic repair of ventral, umbilical, or inguinal hernias. PMID- 9876689 TI - A new modified layout for laparoscopic cholecystectomy. AB - OBJECTIVE: The aim of this study is to present an economic and convenient modification of the layout for laparoscopic cholecystectomy, utilizing a three port technique. METHODS: The surgeon stands on the left side of the patient, while the assistant stands between the patient's legs. The scrub nurse stands on the right side of the patient facing the surgeon. The assistant also operates the camera. Only three ports are used. This technique was used in 119 consecutive patients over a 24-month period. Endoscopic retrograde chlolangiopancreotography (ERCP) was done preoperatively in patients suspected to have choledocholithiasis. RESULTS: Sixteen patients had ERCP done preoperatively and in 12 of them sphincterotomy and stone removal was carried out. Laparoscopic cholecystectomy was successfully completed in 115 patients. The mean operative time was 35 minutes. Four cases were converted (3.6%), one due to bile duct injury, two others due to extensive adhesions, and a fourth due to cholecystoduodenal fistula. The total morbidity rate was 4.2%. The mean hospital stay was 1.8 days. CONCLUSIONS: Laparoscopic cholecystectomy can be safely and conveniently done using only three ports in the modified position described. You need only one assistant, only one monitor and one less trocar. There is no prolongation of the operative time and the results are comparable to the classic four-trocar technique. PMID- 9876690 TI - How managed care may choose hospitals for contracts for laparoscopic cholecystectomy. AB - BACKGROUND AND OBJECTIVES: Maricopa Medical Center (MMC) was found to have higher charges and length-of-stays than 16 other regional hospitals in an analysis of DRG categories for gallbladder disease. These comparative figures identified MMC as being inefficient and demanded review to determine the reasons for the inefficiencies. METHODS: In an attempt to determine the reason for inefficiency of charges and length-of-stay for the laparoscopic portion of laparoscopic cholecystectomy, matched pairs of open cholecystectomy and converted laparoscopic cholecystectomy from a data base of 633 patients with cholecystectomies were reviewed. Thirty-five matches for age, sex and similar diagnosis were successful. RESULTS: Matched pair evaluation disclosed a $6,880 difference in charges, which was attributed solely to the charge for laparoscopy. Subsequent chart analysis showed a high charge for instrumentation, prolonged anesthesia and operative times and longer preoperative delays before surgery. Moreover, no matter what the conversion rate is, open cholecystectomy was more cost effective. However, if there is a conversion rate of 5%, total hospital charges for laparoscopic cholecystectomy would have to be reduced to $12,679 (a reduction of $3,332 from $16,011) to make laparoscopic cholecystectomy cost-effective. CONCLUSIONS: Cost effective decision tree analysis of matched pair comparisons and sensitivity analysis proves to be an effective technique in evaluating the cost-effectiveness of laparoscopic cholecystectomy in a hospital population. PMID- 9876691 TI - Raoul Palmer, World War II, and transabdominal coelioscopy. Laparoscopy extends into gynecology. PMID- 9876692 TI - Best options for cutting and hemostasis during laparoscopic surgery. PMID- 9876693 TI - When is informed consent "informed"? PMID- 9876694 TI - Clinical experience with an optical access trocar in gynecological laparoscopy pelviscopy. AB - BACKGROUND AND OBJECTIVES: Development in surgical technology must demand not only improved efficacy and risk reduction but also a reduction in costs and efficient use of human resources. For 25 years we have discussed the development of optical access trocars and their probable benefits. They are now available in the form of the OPTIVIEW by Ethicon and the SURGIVIEW by US Surgical. METHODS: Between December 1996 and March 1997, we utilized the optic obturator trocar, OPTIVIEW, in 104 cases of gynecological operative laparoscopy. The instrument was equipped with an axial grip to facilitate ergonomic handling. RESULTS: The optical trocar was used with a Z-incision techniques in 16 cases; a vertical incision was used in 58 cases. In all of the gynecological procedures, the optical trocar was more advantageous than classic trocars placed without direct vision. Our estimation was that separation of tissue layers was very good in 71 cases, good in 26 cases and problematic in 5 cases. No complications occurred with the use of this trocar. The Z-incision was preferable to the vertical incision although it required a longer time of insertion of up to 5 seconds. Altogether, the OPTIVIEW presented an easy way of avoiding intestinal and vascular injury during initial trocar entry. CONCLUSIONS: The application of this new tool is practical, safe and handy. However, it requires training in its appropriate use. Vertical incisions should be sutured after removal of the instrument. Additional trocars need not be optical trocars as they can be placed under direct vision and laparoscopic control. It is our opinion that a combination of sophisticated new technologies such as the OPTIVIEW trocar, robotic arm, harmonic scalpel and 3-D vision would provide safe and efficient means to accomplish gynecologic laparoscopic surgical procedures. PMID- 9876695 TI - Effect on negative laparoscopy rate in chronic pelvic pain patients using patient assisted laparoscopy. AB - OBJECTIVE: To determine the value of Patient Assisted Laparoscopy (PAL) in the diagnosis of pelvic pain. METHODS: One hundred patients with pelvic pain were assessed by the procedure of Patient Assisted Laparoscopy to determine the cause of chronic pelvic pain. RESULTS: Of the 100 patients with pelvic pain, 12 patients were not assessed due to technique failure, which included reaction to the carbon dioxide gas, inadequate visualization due to abdominal adhesions or failure to enter peritoneum. Of the remaining 88 patients, 61 had endometriosis; 16 had adhesions not associated with endometriosis; five had hernias; one had occult bowel cancer; one pseudo-stone from previous cholecystectomy; one had pain as a result of staples used at hysterectomy and one patient had chronic Crohn's disease. Two patients had no demonstrated interabdominal cause for their symptoms. CONCLUSION: In contrast to the well published rate of 35% negative laparoscopy in those patients with pelvic pain when examined under general anesthetic, Patient Assisted Laparoscopy decreased the negative laparoscopy rate to less than 3%. This methodology was also of benefit in giving the patient a better understanding of the cause of her pain and the need for therapy. PMID- 9876696 TI - Laparoscopic appendectomy versus open appendectomy: a single institution study. AB - PURPOSE: The purpose of this study was to compare the safety and efficacy of laparoscopic appendectomy versus open appendectomy at Baptist Hospital in Miami, Florida. METHODS: A retrospective review was performed on all appendectomies performed at Baptist Hospital from October 1, 1994 to September 30, 1995. There were a total of 244 cases; 137 open appendectomies and 107 laparoscopic appendectomies. The cases were reviewed with regard to pathology, operating time, length of hospital stay and complications. RESULTS: The pathologic findings at surgery were similar for the two groups. Concomitant pathology was more likely to be found laparoscopically than in open surgery. There was a greater percentage of ruptured appendices in surgery done via the open method. Operative time was slightly longer, but complications were less in the laparoscopic group. Length of stay was lower in the laparoscopic appendectomy group. CONCLUSIONS: Although very similar, our method of appendectomy favors the laparoscopic technique. PMID- 9876697 TI - Microbial colonization of laparoscopic gas delivery systems: a qualitative analysis. AB - OBJECTIVE: Laparoscopic procedures utilize a pneumoperitoneum to distend and separate the abdominal wall from the intra-abdominal structures. Carbon dioxide is commonly used for this purpose, although this study is inclusive of any gas used for abdominal distention. The gas is delivered from cylinders through a gas insufflation delivery system. The purpose of this study is to determine if laparoscopic gas delivery systems composed of gas cylinders and insufflators used for laparoscopy have microbes present. METHODS: Gas delivery systems were evaluated for the presence of microbial growth using standard techniques. External connection sites, gas cylinders and the internal conduit tubing of insufflators were cultured. Fifty two (52) insufflators and sixty (60) gas cylinders were evaluated. RESULTS: Twelve (12) of the sixty cylinders (20%) and fifty four (54) of the sixty insufflators (92.3%) were culture positive. The organisms identified are significant and a varied spectrum. CONCLUSIONS: Recognition that gas cylinders, insufflation attachments and internal components of insufflators quantitatively contain microbes is demonstrated. Reduction of microbial exposure from insufflation apparatus is achieved by cleansing external ports and use of a 0.3 micron filter for abdominal pneumoperitoneum. PMID- 9876698 TI - Image rotation and reversal--major obstacles in learning intracorporeal suturing and knot-tying. AB - BACKGROUND AND OBJECTIVES: A major stumbling block to teaching and learning the finer skills of laparoscopy is related to the "optical illusions" the video camera plays on the surgeon's eyes. Until now, the belief was that lack of coordination was the result of depth perception deficiencies resulting from the two dimensional plane of the video monitor. In reality, this is a minor problem that is easily surmounted with practice. A closer analysis of how organ orientation at the operative site compares to the video camera's fields of focus reveals the real problem: the major optical difference between laparotomy and laparoscopy involves rotation of the images received by the brain. CONCLUSIONS: There are four major operating positions in laparoscopy: camera position, right camera position, left camera position and opposite camera position. The object in front of the camera has two components; the first, a reality image, which results from light reflected off the object as it exists in time and space. The second, a visual image, which represents the actual light entering our eyes. At right camera position the visual image is a 90 degrees counter-clockwise rotation of the reality image. At the left camera position the visual image is a 90 degrees clockwise rotation of the reality image. At opposite camera position, a 180 degrees rotation and complete reversal of the reality image occurs. It is only at camera position that the visual image is equal to the reality image, and we approach a scenario similar to that found in laparotomy. Every other position will be unlike what we were accustomed to in open surgery. PMID- 9876699 TI - Virgin and recurrent groin hernia: a comparison of patient recovery following endoscopic preperitoneal herniorrhaphy. AB - INTRODUCTION: The advantage of minimally invasive hernia repair techniques remains controversial. One of the more established indications for this technique's use is the presence of a recurrent hernia. No prior study has compared the recovery following endoscopic repair of virgin and recurrent hernias. PATIENTS AND METHODS: Between July 15, 1994 through August 16, 1996, one primary surgeon supervised the performance of 373 hernia repairs on 250 patients. Twenty-two patients underwent endoscopic preperitoneal herniorrhaphy for unilateral recurrent groin hernia (RH), while 105 patients underwent repair of a virgin unilateral hernia (VH) in the absence of prior contralateral open hernia repair. No significant differences were seen for age (VH: 54, RH: 64), male:female ratio (VH: 92:13, RH: 22:0), operative time (VH: 58 min, RH: 59 min), anesthetic used, i.v. fluid requirements or blood loss (p > 0.05 for all comparisons). At the time of discharge, all patients were given a postoperative survey and asked to record their level of pain, narcotic use, and level of activity on the day of surgery and postoperative days 1, 2, 3, 7, 14, and 28. RESULTS: Patients undergoing repair of virgin hernias had statistically significant increased levels of pain and/or narcotic use on the day of surgery and postoperative days 1, 2 and 3. Despite these differences, level of activity and return to work/normal activity (VH: 6.35 +/- 3.44 days, RH: 6.40 +/- 2.67 days) were the same for the two groups. CONCLUSION: Despite the differences in pain perception and narcotic use in the early postoperative period, overall patient recovery appears similar for the two groups. Differences seen are likely due to a lack of any prior surgical pain to serve as a benchmark for comparison. PMID- 9876700 TI - Laparoscopically assisted spinal surgery. AB - BACKGROUND: Spinal surgery is one of the newest frontiers of videolaparoscopic surgery, but requires the cooperative efforts of both the spinal surgeon and the laparoscopic general surgeon. DATA BASE: We report our experience with 76 cases of laparoscopic spinal surgery, using both a transperitoneal and a retroperitoneal approach. Technical details and complications are described in detail. CONCLUSIONS: Fifty-one patients had a transperitoneal approach with an average operating time of 117 minutes. Uncomplicated cases stayed 4.4 days. Five patients required conversion. All but one patient had L5-S1 level surgery. Twenty five patients had a retroperitoneal approach with 150 minutes operating time and a 5.7 day stay. Conversions were minimized with a two-balloon technique. The retroperitoneal approach allows for multiple level surgery with virtually unlimited fusion devices. Laparoscopically assisted spine surgery affords all the benefits of minimally invasive surgery, without limitations for the spinal surgeon. PMID- 9876701 TI - Therapeutic endoscopy of the hepatobiliary and pancreatic system: a Vietnamese experience. AB - INTRODUCTION: Therapeutic endoscopic retrograde cholangiopancreatography (ERCP) was initially utilized at Binh Dan Hospital, Viet Nam, in August 1993. From August 1993 through March 1997, 318 ERCP procedures were performed on 271 patients. It was not possible to obtain cholangiography in 32 cases of the 318 procedures of ERCP, for a success rate of diagnostic ERCP approaching 89%. MATERIALS AND METHODS: Cases treated by ERCP included: 14 cases of Ascaris lumbricoides in the common bile duct (CBD). 69 cases of bile duct stones. 12 cases managed by nasobiliary catheter drainage. 3 cases treated by bile duct stent. Sphincterotomy was attempted on 108 cases. Complications included: 5 cases of acute pancreatitis. 7 cases of purulent cholangitis, which resulted in 1 death. 2 cases of retroperitoneal duodenal perforation. 9 cases of postsphincterotomy bleeding. CONCLUSIONS: We conclude that ERCP is a useful therapeutic modality for bile duct stones and parasitic worms in the bile ducts. PMID- 9876702 TI - Laparoscopy in pediatric abdominal trauma. AB - BACKGROUND: The use and indications for laparoscopy have been increasing. As part of this trend, a new algorithm may emerge for pediatric trauma in which laparoscopic techniques are used in hemodynamically stable patients with suspected hollow viscus perforation. CASE REPORT: We present a case in which laparoscopy was successfully used in a pediatric trauma patient as a diagnostic and therapeutic modality. A 4-year-old boy was a back-seat passenger in a head-on collision motor vehicle accident. He was restrained by a lap seat belt. He sustained a concussion, a large forehead laceration and a seat belt abdominal injury. On admission, he complained of abdominal pain. Physical examination revealed a soft, non-distended abdomen with moderate diffuse tenderness. He was hemodynamically stable. Computerized tomography of the abdomen revealed free fluid in the pelvis. No abnormalities were detected in the liver or spleen. Because of clinical deterioration and suspected intestinal perforation, diagnostic laparoscopy was utilized instead of proceeding directly to celiotomy. At laparoscopy a jejunal perforation was found and successfully repaired laparoscopically. Large hematomas were seen in the mesentery, as well as an unsuspected splenic laceration. No active bleeding was found. The patient recovered uneventfully and was discharged 5 days following the surgical procedure. CONCLUSION: This case illustrates the efficacy of using early laparoscopy in children with abdominal trauma when diagnosis is difficult and hollow viscus injury is suspected. PMID- 9876703 TI - Abdominal wall biloma: an unusual complication of laparoscopic cholecystectomy. PMID- 9876704 TI - Hans Frangenheim--culdoscopy vs. laparoscopy, the first book on gynecological endoscopy, and "cold light". AB - In the United States, culdoscopy (a vaginal approach to view the abdomen) replaced laparoscopy for about 20 years, circa 1950-1970. In contrast to many of his colleagues, Hans Frangenheim of Wuppertal, Germany, was not satisfied with culdoscopy and turned to an abdominal approach. Frangenheim began publishing his experiences with gynecological laparoscopy in 1958 and stressed technical improvements. He constructed a CO2 insufflator, wrote the first book on gynecological endoscopy, and introduced "cold light" into laparoscopy. Frangenheim strongly stimulated the rise of gynecological laparoscopy in Europe in the 1960s and later. PMID- 9876705 TI - Laparoscopic wedge resection and partial nephrectomy--the Washington University experience and review of the literature. AB - Open partial nephrectomy is an accepted form of treatment for a variety of benign conditions and for localized renal cell carcinoma. To date, there is limited experience with the clinical application of laparoscopic partial nephrectomy and wedge resection for benign and malignant disease of the kidney. Herein, we report our clinical experience with laparoscopic partial nephrectomy and a review of the current literature. Twelve patients (27-81 years) have undergone laparoscopic wedge resection (3) or attempted polar partial nephrectomy (9) since 1993. In the group of 12 patients, 5 had a mass suspicious for a malignancy, 4 patients had symptomatic polar calyceal dilation with or without stone disease, and 3 patients had an atrophic or hydronephrotic upper pole moiety. Among the patients in the polar nephrectomy group, a third were converted to an open procedure. The remaining 6 patients had a mean operative time of 6.5 hours (5.7-8.3 hours). These patients resumed their oral intake on average 0.8 days postoperatively. In the 2 patients with a mass, the final pathology was oncocytoma (1), and xanthogranulomatous reaction in a renal cyst (1). Postoperative complications included a nephrocutaneous fistula which was endoscopically fulgurated, a retroperitoneal urinoma which was percutaneously drained, and a two-day bout of ileus. The mean hospital stay was 5.3 days (2-9). Their full convalescence was completed in a mean of 4.2 weeks (2-8). Three patients underwent a wedge resection for a superficial < 2 cm mass. The average operative time in this group was 3.5 hours (2-5.4). The mean time to resuming oral intake was 0.7 days (0.3 0.7). The final pathology was oncocytoma (1), oncocytic renal cell cancer (1), and old infarction (1); none of the patients had any complications. The mean hospital stay was 2.7 days (2-4). Convalescence was completed in 4 weeks (range 1 8). Laparoscopic wedge resection and polar partial nephrectomy are feasible, albeit currently tedious techniques. While wedge excision of a < 2 cm superficial lesion is relatively straightforward and efficient, laparoscopic polar partial nephrectomy remains a difficult technique and at present remains in evolution. Further development of instrumentation to provide for a reliable, expeditious, and hemostatic partial nephrectomy is needed. PMID- 9876707 TI - Safe esophageal bougie placement for laparoscopic hiatal hernia repair. AB - The increased use of laparoscopy for treatment of reflux esophagitis has been associated with a 1-8% complication rate. Perforation of the esophagus from bougie placement, wrap breakdown or too tight a wrap are some of the complications seen from this surgery. An esophageal dilator system was developed to overcome these problems. Thirty patients had an esophageal dilator system used whereby a 48 F or 58 F dilator was placed over a 18 F orogastric tube. Intraoperative gastroscopy documented a properly created wrap. There were no esophageal perforations or morbidity associated with the dilator. PMID- 9876706 TI - Analgesia following major gynecological laparoscopic surgery--PCA versus intermittent intramuscular injection. AB - BACKGROUND AND OBJECTIVES: To compare the use of patient-controlled analgesia to intermittent intramuscular injections of morphine following major gynecological laparoscopic procedures in order to assess differences in level of pain, sedation, episodes of nausea and/or vomiting, hospitalization time and patient satisfaction with their postoperative analgesia. METHODS: Seventy-two patients undergoing major gynecological laparoscopic surgery were randomized to receive either postoperative analgesia via intermittent intramuscular injection of morphine (Group 1) or patient controlled analgesia (PCA-Group 2). All patients received anesthesia via a standardized protocol. Postoperative pain levels were recorded via a 10 cm visual analogue scale, and sedation scores were recorded on a standard PCA form. Episodes of nausea and vomiting were also recorded on the same form. RESULTS: There were no statistically significant differences between intramuscular analgesia and PCA for any of the factors studied. Most significantly it was found that most patients ceased to require either form of parenteral analgesia within 24 hours of their procedure, regardless of the operating time. CONCLUSION: It is important for the surgeon to be aware of the effects of postoperative analgesia on his or her patients' level of satisfaction. We do not recommend the use of PCA analgesia following major laparoscopic gynecological surgery. PMID- 9876708 TI - Laparoscopic hernia repair--complications. AB - Laparoscopic hernioplasty is a technique which can present a number of specific complications. This paper reviews the complications that can occur during laparoscopic hernia repair and ways to avoid them; it also describes the surgical technique used successfully in over 1000 cases. Initial experience suggests that complications can be avoided with adequate knowledge, attention to surgical anatomy and the proper technique of laparoscopic hernioplasty. Early recurrences are rare and invariably result from inadequate surgical technique. Inadequate fixation of the mesh, inadequate mesh size, and failure to cover unidentified wall defects (hernias which have never been repaired), are the main causes of early recurrence of hernia. Experience, knowledge of complications and how to avoid them, adequate training and attention to the anatomy of the inguinal region are the most important factors in correcting inguinal hernia successfully by laparoscopy. PMID- 9876709 TI - Laparoscopic appendectomy and cholecystectomy during pregnancy: six case reports. AB - BACKGROUND: Pregnancy was once considered a contraindication to laparoscopic cholecystectomy and appendectomy. The progression of laparoscopic techniques has resulted in a continued reassessment of laparoscopic procedures during pregnancy. There still exists some controversy as to the safety of laparoscopic procedures during pregnancy. This paper reviews our series of six pregnant patients treated laparoscopically for appendicitis and cholecystitis. METHODS: Charts were reviewed of all pregnant patients who underwent laparoscopic cholecystectomy or appendectomy at St Clare's Hospital Schenectady, New York between 1992 and 1996. Six patients were identified. Patients and obstetricians were contacted to investigate the results of the pregnancy. RESULTS: All patients and fetuses survived the procedure. Two patients delivered prematurely but remote from the operative procedure. All infants were healthy postpartum. One patient underwent an elective abortion as she had planned. The abortion was remote from the surgical procedure. CONCLUSION: Our series adds to the growing evidence that laparoscopic cholecystectomy and laparoscopic appendectomy can be performed safely during pregnancy. PMID- 9876710 TI - Facilitating intracorporeal colorectal anastomoses. AB - BACKGROUND AND OBJECTIVES: An experimental study was undertaken to evaluate whether a previously described technique for laparoscopic sigmoid resection with intracorporeal resection-anastomosis and specimen removal via a suprapubic incision could be facilitated and applied to the rectum. METHODS: Ten domestic pigs (median weight 41 kg) underwent low anterior resection of the rectum, which was transected with an articulating endoscopic stapler. Pursestring sutures were fashioned intracorporeally with a laparoscopic pursestring clamp. The anvil of a circular stapler was inserted through a 33 mm port into the colon and pursestring tied intracorporeally. A circular gun with a spike fixed to its shaft was introduced per anum and a double-stapled anastomosis performed. RESULTS: Complete doughnuts were obtained in all cases and anastomoses were all methylene blue tight. All porcine subjects had an uneventful 5-week postoperative course. The median anastomotic level from the anal verge was 5.2 cm. Histology of colorectal anastomoses revealed healing mucosa. CONCLUSIONS: The use of articulating endoscopic stapler, laparoscopic pursestring clamp, and circular stapler with a spike fixed to its shaft seems to facilitate a previously described intracorporeal approach to sigmoid resection which was safely applied to the rectum in a porcine model. PMID- 9876711 TI - Safe and effective fluid management by automated gravitation during hysteroscopy. AB - OBJECTIVE: The automated gravitational Vario Flow system with weighing-based electronic fluid deficit indicator was used in order to reduce the risk of fluid intravasation during continuous flow hysteroscopic procedures. Early experiences are reported. METHODS: Between August 1996 and July 1997, the Vario Flow with fluid deficit indicator and alarm system was used in 203 hysteroscopic operations. Between January 1994 and August 1996 the Vario Flow without fluid deficit indicator was used in 240 hysteroscopic operations. In all, there were 443 hysteroscopic operations: 301 metroplasties, 20 endometrial ablations, 10 cases of lysis of synechiae, 58 myomectomies and 54 polypectomies. The data on fluid deficit before and after the introduction of the electronic fluid deficit indicator were similar. RESULTS: Fluid deficit indicator was proved highly efficient in 203 operations. It provided the information on fluid deficit at any moment during hysteroscopic operations. Besides intrauterine pressure, the actual fluid deficit has become one of the leading parameters during our continuous flow hysteroscopic procedures. CONCLUSIONS: We therefore conclude that by using an automated gravitational system with fluid deficit indicator and alarm system, the safety for patients during hysteroscopic procedures has been increased. PMID- 9876712 TI - Laparoscopic adjustable gastric banding is a safe and effective treatment for morbid obesity. AB - OBJECTIVE: Surgery has been recognized as an effective long-term treatment for morbid obesity. The purpose of this study is to present our experience of laparoscopic adjustable gastric banding (LAGB) as a safe and effective treatment for morbid obesity. METHODS AND PROCEDURES: Over eight months, 39 morbidly obese patients, having a Body Mass Index (BMI) > 40 kg/m2, were included in this study. Conservative measures failed to maintain weight loss in all patients. The procedure is performed through a 5-trocar technique. The procedure involves gastric partitioning and stoma formation by an inflatable band. The stoma can be adjusted by injection of saline in the band reservoir. RESULTS: The mean age of the patients was 31.3 years. The mean BMI was 44.2 Kg/m2. All procedures were completed laparoscopically. The mean hospital stay was 2.7 days. The morbidity rate was 15.32%. Patients were followed up for a mean period of 6.7 months. The mean BMI after six months (in 28 patients) was 36.6 Kg/m2. CONCLUSION: Laparoscopic insertion of the adjustable gastric banding is a safe and effective method for the treatment of morbid obesity and should be the standard way of band insertion. PMID- 9876713 TI - Laparoscopic transverse rectus abdominus flap delay for autogenous breast reconstruction. AB - Laparoscopic ligation of the deep and superficial inferior epigastric vessels was done for ten mastectomized patients who elected to have autogenous reconstruction of their breast. All these patients had at least one indication for the delay which included obesity, smoking, or requirement of a large volume of tissue for their reconstruction. The procedure did not add any morbidity or mortality to our patients and was found to be comparable to the "open" delay in preventing partial tissue loss in all but two patients. We describe the use of a minimally invasive procedure to augment the deep superior epigastric pedicled blood supply for the future transverse rectus abdominus flap. We have found in laparoscopic delay a safe, short procedure that is useful in high risk patients who choose the option of autologous breast reconstruction. PMID- 9876714 TI - A simple stereoscopic endoscope. AB - A very simple method is described for producing and viewing stereoscopic endoscopic images. The addition of two simple prisms to the end of a conventional television-monitored endoscope with a simple viewing device produces a stereoscopic endoscope which appears to be suitable for surgical use. PMID- 9876715 TI - The "floppy diaphragm" sign with laparoscopic-associated pneumothorax. AB - Pneumothoraces in association with laparoscopy are uncommon and potentially disastrous complications that may also occur without adverse sequelae. The "floppy diaphragm sign" is a readily discernible and useful sign of a laparoscopic-induced pneumothorax. Tube thoracostomy is generally not indicated in stable patients as the pneumothorax typically resolves quickly upon desufflation of the pneumoperitoneum. PMID- 9876716 TI - Laparoscopic transhiatal esophagectomy for Barrett's esophagus with high grade dysplasia. AB - BACKGROUND: A number of case reports have described the application of minimally invasive surgical techniques to accomplish esophagectomy. However, most reports have employed thoracoscopic or laparoscopic techniques to perform esophagectomy in addition to an "access" incision which often approaches a standard laparotomy or thoracotomy. CASE REPORT: This report describes a total laparoscopic transhiatal esophagectomy in a 55 year old female with Barrett's esophagus and high grade dysplasia. CONCLUSIONS: The patient had an uneventful recovery and was discharged home on the fourth day after a total laparoscopic esophagectomy. This report demonstrates the technical feasibility of this complex procedure by a minimally invasive approach. PMID- 9876718 TI - Posterior wall gastric leiomyoma: endoscopic tattooing facilitates laparoscopic resection. AB - OBJECTIVE: To demonstrate the application of tattooing for the intraoperative localization of posterior wall gastric leiomyoma during laparoscopic resection. The preoperative injection of Indian ink in the tumor-bearing area of the posterior gastric wall eliminates the need to perform anterior wall gastrostomy or intraoperative upper endoscopic tumor localization. METHODS: A patient with posterior wall gastric leiomyoma was marked with Indian ink during preoperative upper endoscopy. The dye was visualized intraoperatively facilitating wedge resection of the tumor-bearing area with the Endo GIA. RESULTS: The patient had an uneventful surgery and recovery. Complete excision of the tumor was accomplished. CONCLUSION: The preoperative endoscopic marking of gastric lesions, facilitates the intraoperative localization and resection of these lesions. PMID- 9876717 TI - Minilaparoscopy without general anesthesia for the diagnosis of acute appendicitis. AB - The timely diagnosis of intra-abdominal pathology continues to be an elusive problem. Delays in diagnosis and therapeutic decision making are continuing dilemmas in patients who are females of childbearing age, elderly, obese or immunosuppressed. Minilaparoscopy without general anesthesia potentially can provide an accurate, cost-effective method to assist in the evaluation of patients with acute abdominal pain. Laparoscopy without general anesthesia is not a new technique, but with the combination of two emerging factors--1) the introduction of new technology with the development of improved, smaller laparoscopes and instruments, and 2) the shifting of emphasis on healthcare to a more cost-effective managed care environment--its value and widespread utilization is being reconsidered. We report the case of a 22 year old female with an acute onset of increasing abdominal and pelvic pain. Despite evaluation by general surgery, gynecology, emergency room staff, as well as, non-invasive testing, a clear diagnosis could not be made. In view of this, minilaparoscopy without general anesthesia was performed and revealed an acute, retrocecal appendicitis. The diagnosis was made with the assistance from the conscious patient. The utilization of this technique greatly expedited the treatment of this patient. Full-sized laparoscopic equipment was then used to minimally invasively remove the diseased appendix under general anesthesia. Both procedures were well tolerated by the patient. PMID- 9876719 TI - The laparoscopic approach for repair of Morgagni hernias. AB - BACKGROUND AND OBJECTIVES: Morgagni hernias are unusual congenital diaphragmatic hernias that are generally asymptomatic and discovered incidentally. Surgical treatment is indicated once the diagnosis is made. These hernias have traditionally been repaired by the open abdominal or thoracic approaches. We report a case of Morgagni hernia repaired successfully via the laparoscopic approach. METHODS AND RESULTS: The patient was noted to have a large anteromedial diaphragmatic hernia by chest radiograph and CT imaging. He underwent laparoscopy, during which the hernia was reduced and the defect repaired with mesh placement. We used intracorporeal suture placement to anchor the mesh. The patient recovered uneventfully after a short hospitalization. CONCLUSIONS: The laparoscopic approach for repair of Morgagni hernias offers diagnostic advantages as well as the potential for reduced morbidity when compared to laparotomy. We report intracorporeal knot-tying for fixation of the mesh to be a secure and satisfactory means to achieve the laparoscopic repair. PMID- 9876720 TI - Laparoscopic cholecystectomy in a cardiac transplant candidate with an ejection fraction of less than 15%. AB - BACKGROUND AND OBJECTIVES: Laparoscopic procedures are becoming increasingly popular, even in the severely ill patient. We present a case of a cardiac transplant candidate with an ejection fraction of less than 15% who underwent a laparoscopic cholecystectomy. This is the first case in which intraoperative hemodynamic measurements were recorded in a patient with cardiopulmonary disease this severe. METHODS: The patient underwent the procedure while measurements were made at crucial intervals (baseline, with incremental insufflation, reverse Trendelenberg, at desufflation) using a pulmonary artery catheter, transesophageal echocardiography with fractional area measurements, radial arterial line, as well as standard monitoring. RESULTS: This patient showed hemodynamic changes consistent with a person without severe heart disease. There was a rise in mean arterial and systemic vascular resistance with insufflation to 10 mm Hg, which was further exaggerated by reverse Trendelenberg. Heart rate and pulmonary artery wedge pressure increased slightly. The cardiac output and fractional area change declined minimally. CONCLUSIONS: We conclude that if a patient with congestive heart failure is medically optimized, and intra-abdominal pressures and surgical times are minimized, laparoscopic cholecystectomy may be performed with minimal risk to the patient. PMID- 9876721 TI - Percutaneous endoscopic treatment of bleeding gastric ulcers. AB - Bleeding gastric ulcers have a propensity to occur in the critically ill and elderly. Various treatments include endoscopic or surgical intervention. The endoscopic route may have as high as a 25 percent rebleeding rate and a 50 to 90 percent rebleeding rate if there is a visible vessel. Surgical intervention has a mortality rate of as high as 25 percent. Described is a procedure which combines endoscopic and surgical techniques for the treatment of bleeding gastric ulcers- the percutaneous endoscopic method. To evaluate the technique and efficacy of this procedure, a bleeding ulcer was created in a porcine stomach with multiple applications of standard biopsy forceps. After brisk hemorrhage developed, the percutaneous endoscopic method was employed and cessation of bleeding obtained. The entire procedure was completed within 20 minutes. There were no immediate complications. Overall costs were much less than those for routine general surgery and about the same as for placement of percutaneous gastrostomy tube. PMID- 9876722 TI - Laparoscopic access to the preperitoneal space. PMID- 9876723 TI - Patrick C. Steptoe: laparoscopy, sterilization, the test-tube baby, and mass media. AB - In the late 1950s, Patrick C. Steptoe, a British gynecologist, established contact with Palmer of Paris and Frangenheim of Wuppertal, Germany, and studied laparoscopic technique under the tutelage of these pioneers. Despite the negative attitude among his colleagues, Steptoe soon became one of the most innovative researchers in the field of abdominal endoscopy, particularly laparoscopic sterilization. In the late 1960s, Steptoe began working with Robert Edwards, an embryologist, and launched an in-vitro fertilization project obtaining eggs by means of laparoscopy. Both researchers experienced years of frustration, disappointment, ethical and scientific criticism as well as a difficult relationship with the mass media. Finally, in July 1978, Louise Brown, the first test-tube baby, was born in England. Like many of his colleagues in the 1950s and 1960s, Patrick Christopher Steptoe (1913-1988), a gynecologist in Oldham, Great Britain, was concerned about the number of unnecessary laparotomies. Unfortunately, the Oldham group of hospitals was not a university clinic and Steptoe had scanty opportunity to develop his own research. In the late 1950s, he searched the medical literature for an alternative form of examination and came across publications about Decker's culdoscopy, the vaginal approach to view of the abdomen. Since this method was not widespread in England, Steptoe, in 1958, went to Montreal, Boston, and New York in order to observe and learn the practical use of culdoscopy. However, Steptoe left America disappointed. PMID- 9876724 TI - Guidelines for an office-based surgical facility: quality not bureaucracy. PMID- 9876725 TI - Laparoscopy for penetrating thoracoabdominal trauma: pitfalls and promises. AB - BACKGROUND: How should the stable patient with penetrating abdominal or lower chest trauma be evaluated? Evolving trends have recently included the use of diagnostic laparoscopy. In September 1995 we instituted a protocol of diagnostic laparoscopy to identify those patients who could safely avoid surgical intervention. DESIGN: Prospective case series. MATERIALS AND METHODS: Hemodynamically stable patients with penetrating injuries to the anterior abdomen and lower chest were prospectively evaluated by diagnostic laparoscopy, performed in the operating room under general anesthesia, and considered negative if no peritoneal violation or an isolated nonbleeding liver injury had occurred. If peritoneal violation, major organ injury or hematoma was noted, conversion to open celiotomy was undertaken. RESULTS: Seventy consecutive patients were evaluated over a two-year period. The average length of stay (LOS) following negative laparoscopy was 1.5 days, and for negative celiotomy 5.2 days. There were no missed intra-abdominal injuries following 30 negative laparoscopies, and 26 of 40 laparotomies were therapeutic. The technique also proved useful in evaluation of selected blunt and HIV+ trauma victims with unclear clinical presentations. However, while laparoscopy was accurate in assessing the abdomen following penetrating lower chest injuries, significant thoracic injuries were missed in 2 out of 11 patients who required subsequent return to OR for thoracotomy. CONCLUSIONS: Laparoscopy has become a useful and accurate diagnostic tool in the evaluation of abdominal trauma. Nevertheless, laparoscopy still carries a 20% nontherapeutic laparotomy rate. Additionally, significant intrathoracic injuries may be missed when laparoscopy is used as the primary technique to evaluate penetrating lower thoracic trauma. PMID- 9876726 TI - Surgical treatment for chronic pelvic pain. AB - The source of chronic pelvic pain may be reproductive organ, urological, musculoskeletal-neurological, gastrointestinal, or myofascial. A psychological component almost always is a factor, whether as an antecedent event or presenting as depression as result of the pain. Surgical interventions for chronic pelvic pain include: 1) resection or vaporization of vulvar/vestibular tissue for human papillion virus (HPV) induced or chronic vulvodynia/vestibulitis; 2) cervical dilation for cervix stenosis; 3) hysteroscopic resection for intracavitary or submucous myomas or intracavitary polyps; 4) myomectomy or myolysis for symptomatic intramural, subserosal or pedunculated myomas; 5) adhesiolysis for peritubular and periovarian adhesions, and enterolysis for bowel adhesions, adhesiolysis for all thick adhesions in areas of pain as well as thin ahesions affecting critical structures such as ovaries and tubes; 6) salpingectomy or neosalpingostomy for symptomatic hydrosalpinx; 7) ovarian treatment for symptomatic ovarian pain; 8) uterosacral nerve vaporization for dysmenorrhea; 9) presacral neurectomy for disabling central pain primarily of uterine but also of bladder origin; 10) resection of endometriosis from all surfaces including removal from bladder and bowel as well as from the rectovaginal septal space. Complete resection of all disease in a debulking operation is essential; 11) appendectomy for symptoms of chronic appendicitis, and chronic right lower quadrant pain; 12) uterine suspension for symptoms of collision dyspareunia, pelvic congestion, severe dysmenorrhea, cul-desac endometriosis; 13) repair of all hernia defects whether sciatic, inguinal, femoral, Spigelian, ventral or incisional; 14) hysterectomy if relief has not been achieved by organ-preserving surgery such as resection of all endometriosis and presacral neurectomy, or the central pain continues to be disabling. Before such a radical step is taken, MRI of the uterus to confirm presence of adenomyosis may be helpful; 15) trigger point injection therapy for myofascial pain and dysfunction in pelvic and abdominal muscles. With application of all currently available laparoscopic modalities, 80% of women with chronic pelvic pain will report a decrease of pain to tolerable levels, a significant average reduction which is maintained in 3 year follow-up. Individual factors contributing to pain cannot be determined, although the frequency of endometriosis dictates that its complete treatment be attempted. The beneficial effect of uterosacral nerve ablation may be as much due to treatment of occult endometriosis in the uterosacral ligaments as to transection of the nerve fibers themselves. The benefit of the presacral neurectomy appears to be definite but strictly limited to midline pain. Appendectomy, herniorraphy, and even hysterectomy are all appropriate therapies for patients with chronic pelvic pain. Even with all laparoscopic procedures employed, fully 20% of patients experience unsatisfactory results. In addition, these patients are often depressed. Whether the pain contributes to the depression or the depression to the pain is irrelevant to them. Selected referrals to an integrated pain center with psychologic assistance together with judicious prescription of antidepressant drugs will likely benefit both women who respond to surgical intervention and those who do not. A maximum surgical effort must be expended to resect all endometriosis, restore normal pelvic anatomy, resect nerve fibers, and treat surgically accessible disease. In addition, it is important to provide patients with chronic pelvic pain sufficient psychologic support to overcome the effects of the condition, and to assist them with underlying psychologic disorders. PMID- 9876727 TI - Videolaparocholecystectomy: casuistry of 1000 cases. AB - Despite the ongoing evolution in the medical treatment of biliary pathology, the standard of treatment for gallstones remains cholecystectomy. There are no alternative treatments that have shown the same efficacy as surgery. Current alternative treatments have shown high recurrence and failure rates. Cholecystectomy remains the gold standard for management of gallstones. The surgical access of laparoscopic cholecystectomy accomplished by Mouret in 1987 allows for a reduction in operative trauma, hospital stay, postoperative pain and convalescence. These factors permit a faster return to normal activities. Today, laparoscopic cholecystectomy is performed in almost all medical centers around the world; however, the procedure is not free of complications. The objective of this study was to analyze our first 1000 cases of laparoscopic cholecystectomy, giving emphasis to the morbidity and mortality of the procedure. PMID- 9876728 TI - A multidisciplinary approach to major bile duct injury following laparoscopic cholecystectomy. AB - BACKGROUND AND OBJECTIVES: Many series describing the management of major bile duct injuries after laparoscopic cholecystectomy have been reported with satisfactory short-term results. However, the information of their prognosis with sufficient time-period follow-up is sparse. METHODS: Sixteen consecutive patients with major bile duct injury following laparoscopic cholecystectomy were retrospectively reviewed, including six common bile duct transections, four bile duct perforations, and six hilar strictures but without perforation. With respect to the level of bile duct injuries, there were the following based on Bismuth's classification: type 1 in six patients, type 2 in five patients, type 3 in three patients, type 4 in one patient, and type 5 in one patient. All patients received surgical management, interventional radiology and endoscopic treatment. The time periods of follow-up ranged from 37 to 72 months (mean, 52 months). The final results were rated as being excellent, good, fair, or poor, based on the criteria of symptoms, biochemical data, and radiology. RESULTS: There was no procedure related mortality. Ten of the 16 patients had either excellent or good results, two had fair results, and four had poor results. Of the latter four, the patients had been classified as Bismuth type 1, 3, 4, and 5, respectively, and all sustained a failed initial surgical repair. CONCLUSIONS: Using a multidisciplinary approach, 12 (75%) of the 16 patients attained a promising result through a long-term follow-up, while those with the higher biliary stricture and with an unsuccessful initial surgical repair had a disappointing outcome. PMID- 9876729 TI - A comparison of laparoscopic and open appendectomy. AB - BACKGROUND AND OBJECTIVES: To compare laparoscopic appendectomy with traditional open appendectomy. METHODS: Seventy-one patients requiring operative intervention for suspected acute appendicitis were prospectively compared. Thirty-seven patients underwent laparoscopic appendectomy, and 34 had open appendectomy through a right lower quadrant incision. Length of surgery, postoperative morbidity and length of postoperative stay (LOS) were recorded. Both groups were similar with regard to age, gender, height, weight, fever, leukocytosis, and incidence of normal vs. gangrenous or perforated appendix. RESULTS: Mean LOS was significantly shorter for patients with acute suppurative appendicitis who underwent laparoscopic appendectomy (2.5 days vs. 4.0 days, p < 0.01). Mean LOS was no different when patients classified as having gangrenous or perforated appendicitis were included in the analysis (3.7 days vs. 4.1 days, P = 0.11). The laparoscopy group had significantly longer surgery times (72 min vs. 58 min, p < 0.001). There was no significant difference in the incidence of postoperative morbidity. CONCLUSIONS: Laparoscopic appendectomy reduces LOS as compared with the traditional open technique in patients with acute suppurative appendicitis. The longer operative time for the laparoscopic approach in our study is likely related to the learning curve associated with the procedure and did not increase morbidity. PMID- 9876730 TI - The laparoscopic approach in the treatment of diverticular colon disease. AB - BACKGROUND AND OBJECTIVES: The experience with treatment of diverticular colon disease (DCD) by the laparoscopic method is analyzed. METHODS: Between January 1994 and July 1997, a group of 22 patients with criteria for symptomatic diverticular disease in the descending and sigmoid colon underwent laparoscopy with average resections of 40 cm. Intra-abdominal mechanical anastomosis completed the procedure. RESULTS: The operative morbidity was 28%. Two cases, in acute diverticulitis phase, were reconverted to open surgery, and three cases presented postoperative rectorrhagia which ceased spontaneously. No long-term complications have been found. Postoperative hospitalization was 4-8 days (mean 5.5) and mean operative time was 165 minutes (range 120-240). CONCLUSIONS: Nevertheless, the learning curve precise to practice this type of surgery, the acceptable morbity-mortality rates which the laparoscopic method presents, especially with these high-risk groups of patients (age > 65, high blood pressure, etc), encouraged us to modified the criteria indicating surgery for the disease, offering first choice operative treatment with efficiency and safety. However, we feel that those patients with acute complications of diverticular colon disease must be excluded initially for laparoscopic approach. PMID- 9876731 TI - Correlation of the end-tidal PCO2 during laparoscopic surgery with the pH of the gastric juice. AB - BACKGROUND AND OBJECTIVES: During laparoscopy, the increase of the carbon dioxide tension may increase the synthesis of hydrochloric acid in the parietal cells of the stomach; the source of the secreted hydrogen ions is carbonic acid derived from the hydration of carbon dioxide. The present report tests this hypothesis by correlating the changes of end-tidal PCO2 (ETCO2) with the pH of the gastric juice in patients undergoing laparoscopic cholecystectomy. METHODS: 40 adult patients were investigated: 20 controls, and 20 patients receiving 100 mg nizatidine intravenously, prior to surgery. In both groups, the ETCO2 was measured by capnography and the pH of the gastric juice was monitored before carbon dioxide insufflation and at the end of laparoscopy prior to carbon dioxide deflation. RESULTS: In the control group, the ETCO2 increased following carbon dioxide insufflation from a mean basal value of 30.2 (standard deviation [SD] 4.6) mm Hg to 41.1 (SD 9.5) mm Hg, while the mean pH of the gastric juice decreased significantly from 1.9 (SD 0.4) to 1.27 (SD 0.43). There was a significant negative correlation between the ETCO2 and pH of the gastric juice (r = -0.4). In the Nizatidine group, the ETCO2 also increased following carbon dioxide insufflation from a mean basal value of 30.9 (SD 3.0) mm Hg to 39.4 (SD 5.3) mm Hg. However, in contrast with the control group, the mean pH of the gastric juice did not decrease, but paradoxically increased from 1.68 (SD 0.36) to 3.6 (SD 1.02). CONCLUSIONS: During laparoscopy, the pH of the gastric juice is significantly decreased. This decrease is inversely related to the increase of ETCO2. The preoperative administration of the selective H2-blocker nizatidine can prevent the increase in gastric acidity and can result in a paradoxical increase of pH of the gastric juice. PMID- 9876732 TI - The role of laparoscopy in the management of groin hernia. AB - INTRODUCTION: The advantage of using minimally invasive techniques over open techniques in the repair of groin hernias is still debated. Despite its more widespread use, an apparent dichotomy exists. While some surgeons continue to believe that no advantage is gained using the laparoscopic technique, others argue laparoscopic hernia repair (LHR) offers a quicker recovery with the use of a tension-free repair. METHODS: A mailing to the general surgeon members of the Society of Laparoendoscopic Surgeons, an international multidisciplinary laparoendoscopic society, was performed (mailing size = 1680). RESULTS: Nine hundred and ninety-three surgeons responded (60%). Across all demographic variables, 60% of respondents performed approximately 27% of their hernia repairs laparoscopically (40% of respondents did not perform LHR). Surgeon age less than 45 was the only demographic characteristic that predicted the likelihood to perform LHR (p < 0.0001) and the percentage of hernias repaired laparoscopically (p < 0.005). Most respondents felt that the presence of bilateral hernias (73%) or a recurrent hernia (74%) were indications for LHR. Eighty-nine percent of respondents felt that LHR would still be performed 20 years from now. Surgeons expressed concerns regarding increased cost, the need for more anesthesia, and a lack of long-term follow-up for LHR. CONCLUSIONS: Only surgeon age predicted the likelihood of a surgeon performing LHR or the percentage of hernias that would be repaired laparoscopically. PMID- 9876733 TI - Laparoscopic re-do repairs of recurrent inguinal hernias using double-mesh technique. AB - Described is a "double mesh" technique for performing laparoscopic re-do repairs of inguinal hernias. When doing this procedure, it is virtually impossible to take down the peritoneum due to incorporation of the old mesh. This technique is therefore done by using a simple onlay of polypropylene mesh, covering the hernia defect and ensuring that sufficient staples are placed into the iliopubic tract. Proper recognition of neuroanatomy is essential. In order to prevent intestinal adhesions, a second patch of gortex is secured to the polypropylene. PMID- 9876734 TI - Ventriculoperitoneal shunt failure as a complication of laparoscopic surgery. AB - OBJECTIVE: The authors report the first documented case of laparoscopically induced ventriculoperitoneal (VP) shunt failure. SUMMARY BACKGROUND DATA: Laparoscopic surgery has become a preferred method of accessing and treating a variety of intraperitoneal pathology. Surgeons can expect to encounter patients who have previously undergone placement of cerebrospinal fluid (CSF) shunts who present as candidates for laparoscopic procedures. Currently, the presence of a CSF shunt is not considered to be a contraindication to laparoscopy. We report the first documented case of laparoscopically induced VP shunt failure. CLINICAL HISTORY: A patient with shunt-dependent hydrocephalus underwent laparoscopic placement of a feeding jejunostomy. Postoperatively, clinical and radiographic evidence of shunt failure was noted. The patient underwent emergent shunt revision. Intraoperatively, an isolated distal shunt obstruction was encountered. Gentle irrigation cleared the occlusion. We believe that this shunt dysfunction was secondary to impaction of either soft tissue or air within the distal catheter as a consequence of peritoneal insufflation. CONCLUSIONS: It is concluded that laparoscopic surgery may represent a potential danger in patients with pre-existing CSF shunts. The risk of neurological injury faced by this patient population during laparoscopy is derived from peritoneal insufflation and relates to two primary concerns. The first is impaired CSF drainage due to a sustained elevated distal pressure gradient or, as in our case, an acute distal catheter obstruction. The second concern relates to the potential for retrograde insufflation of the CSF spaces through an incompetent shunt valve mechanism. Distal shunt catheter externalization performed in conjunction with a neurosurgeon during the laparoscopic procedure would prevent these complications. Internalization of the distal shunt catheter would then be performed at the completion of the laparoscopic procedure. PMID- 9876735 TI - Laparoendoscopic diagnosis of eosinophilic enteritis. AB - As the popularity of laparoscopic cholecystectomy continues to grow, evaluation of patients with documented cholelithiasis and concomitant vague abdominal complaints becomes less rigorous. We present the case of a patient with chronic cholecystitis documented by history and ultrasonography, incidentally noted on laboratory examination to have peripheral blood eosinophilia. At the time of laparoscopy, an inflamed segment of jejunum was discovered. Limited laparotomy and wedge biopsy revealed active eosinophilic enteritis. PMID- 9876736 TI - Gastric outlet obstruction secondary to post cholecystectomy biloma: case report and review of the literature. AB - BACKGROUND: Postcholecystectomy bilomas are relatively uncommon with a reported incidence of about 2.5%, and most often present with right upper quadrant pain and fever within seven days of the operation. There are a number of approaches to the treatment of this uncommon lesion. CASE REPORT: The authors report a case of a cholecystectomy performed in a 35-year-old female which resulted in a biloma, presenting 11 days postoperatively with gastric outlet obstruction. The literature is reviewed to suggest the optimal management and treatment plans for this complication. RESULTS: This patient was successfully managed with radiologic intervention for the treatment of a biloma and resulting gastric outlet obstruction after open cholecystectomy. CONCLUSION: Although uncommon, bilomas may present in an unusual manner such as with gastric outlet obstruction. As with early bilomas, they may be treated successfully with interventional radiologic techniques. PMID- 9876737 TI - Laparoscopically assisted surgery for colonic perforation with peritonitis--a case report. AB - Elective laparoscopic colonic surgery is increasingly recognized as feasible and perhaps preferential. A case of laparoscopically assisted surgery for trauma to the rectum with bacterial peritonitis is presented. It presents an example of the application of this modality to the treatment of iatrogenic colon perforations and perhaps selected diverticulitis. PMID- 9876738 TI - Obturator hernia: laparoscopic diagnosis and repair. AB - OBJECTIVE: Review of international literature reveals eight reported cases of laparoscopic obturator hernia repair. Non-specific signs and symptoms make the diagnosis of an obturator hernia difficult. Laparoscopic intervention provides a minimally invasive method to simultaneously diagnose and repair these hernias. METHODS AND PROCEDURES: A 35 year old woman presented with lower abdominal pain, vaginal bleeding, and dyspareunia. During gynecological diagnostic laparoscopy, a pelvic floor hernia was suspected, and a general surgical evaluation was sought. At a subsequent laparoscopy, the diagnosis of a left direct inguinal and a right obturator hernia was made. Both were repaired laparoscopically with polypropylene mesh. RESULTS: At follow-up at one and six weeks postoperatively, the patient's complaints of pain had completely resolved. CONCLUSION: The diagnosis of obturator hernia is problematic. The usual presenting signs and symptoms are non specific. Without conclusive historical or physical findings, laparoscopy is an excellent method for diagnosing obturator hernia. This entity, once diagnosed laparoscopically, can be repaired simultaneously via laparoscopic mesh technique. PMID- 9876739 TI - Conservative treatment of recognized laparoscopic colonic injury. AB - Injuries associated with insufflation needles and trocar insertion have been reported extensively in the literature. Two millimeter laparoscopy is a more recent technique that has been used for laparoscopic cholecystectomy. This case illustrates a 2 mm trocar colonic injury, recognized during a routine laparoscopic cholecystectomy; management was non-operative and ambulatory, with a successful outcome. PMID- 9876740 TI - Kurt Semm and an automatic insufflator. AB - Work on tubal insufflation marked the beginning of Kurt Semm's (b. 1927) scientific career. In the early 1960s, he directed his attention to the fact that, from a technical standpoint, tubal insufflation was similar to creating pneumoperitoneum. In the mid-1960s, Semm--himself a gynecologist--investiged his time and financial resources and risked his university career to develop an automatic abdominal insufflation device. Later he tried it out in the Clinic for Internal Medicine. Since, at that time, the term "laparoscopy" had negative connotations associated with it, Semm formulated a new term "pelviscopy." In 1967, Semm presented his invention to Melvin Cohen, an American pioneer of gynecological laparoscopy, at the meeting of the American Fertility Society, held in Washington. PMID- 9876741 TI - A new modified layout for laparoscopic cholecystectomy. PMID- 9876742 TI - The World Congress. PMID- 9876743 TI - Patient safety during laparoscopic monopolar electrosurgery--principles and guidelines. Consortium on Electrosurgical Safety During Laparoscopy. PMID- 9876744 TI - Minimally traumatic techniques for in utero access and fetal surgery. AB - BACKGROUND AND OBJECTIVES: Prenatal interventions may prevent some sequelae of congenital anomalies, yet open fetal surgery is limited by pre-term labor. We are developing amnioscopic strategies to reduce risks for in utero surgery. METHODS: Seven fetal sheep were accessed percutaneously under ultrasound guidance, via maternal laparoscopy/transuterine trocars, or via laparoscopically assisted partial uterine exteriorization (mini-hysterotomy). Three fetal immobilization methods were investigated: 2 mm grasping forceps, 2-0 absorbable Roeder loop lasso, and detachable metal T-fasteners. RESULTS: Percutaneous access with 2 mm trocars required accessible amniotic fluid pockets. The laparoscopically assisted method enabled 5 mm trocar placement under direct fetal visualization. Mini hysterotomy avoided trocars and was leak-proof. 2 mm graspers enabled fetal manipulation with repeatable applications. Roeder loop sutures were not readily repositioned, and required two 3 mm or larger trocars. T-fasteners were easily deployed into fetal skin via the self-contained needle applicator with minimal hemorrhage. Cutaneous marks were present immediately from the grasper and T fastener, but not at postoperative day 10. The Roeder loop produced no observable effects. All devices demonstrated adequate intrauterine performance. CONCLUSIONS: Minimally invasive fetal surgery promises to lower maternal-fetal risks. All strategies permitted prolonged amnioscopy and fetal manipulation. The 2 mm grasper was easiest to use, producing no observable lasting trauma. PMID- 9876745 TI - Non-gynecologic laparoscopy in second and third trimester pregnancy: obstetric implications. AB - PURPOSE: To review the effect of non-gynecologic laparoscopic procedures performed during the second and third trimesters of pregnancy on pregnancy outcome. MATERIALS AND METHODS: A review of the patient log for the antenatal obstetrical unit was used to identify the patients in this series from January 1, 1997 to December 31, 1997. Medical records were then analyzed to identify estimated gestational age at surgery and delivery, type of delivery, use of tocolysis, and complications from surgery. RESULTS: Nine patients were identified as having non-gynecologic laparoscopic surgery (without conversion to laparotomy) during the second or third trimester of pregnancy. The median estimated gestational age at surgery was 25 weeks (mean 24 weeks). The most common procedure performed was laparoscopic cholecystectomy (6 patients). Five patients received tocolysis after the initial procedure. All patients delivered at greater than or equal to 37 weeks estimated gestational age (median 38 weeks). No infants were admitted to the neonatal intensive care unit. CONCLUSIONS: Laparoscopic procedures appear safe in second and third trimester pregnancy. In this study, laparoscopic cholecystectomies were performed as late as 34 weeks estimated gestational age without any adverse effects on pregnancy outcome. PMID- 9876746 TI - Laparoscopic surgery for anorectal malignancies other than carcinoma. AB - OBJECTIVE: Numerous concerns have been raised relative to the appropriateness of laparoscopic surgery for cure of rectal adenocarcinomas. However, because of their rarity, little information exists about the role of laparoscopy for other anorectal malignancies. We report the outcome of five patients who underwent laparoscopic surgery for other anorectal malignancies. METHODS: All patients who underwent laparoscopic surgery for treatment of non-carcinomatous anorectal malignancy were assessed by means of endoscopic, radiological and histopathologic diagnostic tests. RESULTS: Two patients with anorectal melanoma and one with anal leiomyosarcoma underwent laparoscopic abdominoperineal resection. A laparoscopic loop ileostomy was performed for an HIV-positive patient with rectal Kaposi's sarcoma. Another patient with anorectal melanoma had intraoperative identification of distant liver metastasis and therefore underwent diagnostic laparoscopy instead of an intended abdominoperineal resection. There were no intraoperative laparoscopic complications. During the follow-up period three patients who underwent abdominoperineal resection were alive, one of whom had rectal melanoma and developed liver metastasis without local recurrence. The two patients with distant liver metastasis and rectal Kaposi's sarcoma died 46 days and five months after surgery, respectively. There were no port-site or local recurrences. CONCLUSION: Laparoscopic abdominoperineal resection for non carcinomatous anorectal malignancies is technically feasible and avoids many of the concerns associated with attempted curative laparoscopic resection of carcinoma. PMID- 9876747 TI - Minimally invasive approach to esophagectomy. AB - BACKGROUND: Recent advances in laparoscopic and thoracoscopic surgery have made it possible to perform esophagectomy using minimally invasive techniques. The aim of this report was to present our preliminary experience with minimally invasive esophagectomy. METHODS: We reviewed our experience on eight patients who underwent minimally invasive esophagectomy using either laparoscopic and/or thoracoscopic techniques from June 1996 to May 1997. Indications for esophagectomy included stage I carcinoma (5), palliative resection (1), Barrett's with high grade dysplasia (1) and end stage achalasia (1). RESULTS: The average age was 68 years (54-82). The surgical approach to esophagectomy included laparoscopic transhiatal esophagectomy with cervical anastomosis (n = 4), thoracoscopic and laparoscopic esophagectomy with cervical anastomosis (n = 1), and laparoscopic mobilization with right mini-thoracotomy and intra-thoracic anastomosis (n = 3). Conversion to mini-laparotomy was required in two patients (25%) to complete esophageal dissection and facilitate gastric pull-up. The mean operative time was 460 minutes. The mean intensive care stay was 1.9 days (range of 0-7 days) with a mean hospital stay of 13.8 days. Minor complications included atrial fibrillation (n = 1), pleural effusion (n = 2) and persistent air leak (n = 1). Major complications included cervical anastomotic leak (n = 1), and delayed gastric emptying requiring pyloroplasty (n = 1). There was no perioperative mortality. CONCLUSIONS: This preliminary experience suggests that minimally invasive esophagectomy is safe and feasible in centers with experience in advanced minimally invasive surgical procedures. Further studies are necessary to determine advantages over open esophagectomy. PMID- 9876748 TI - Thoracoscopic sympathectomy for vasospastic diseases. AB - BACKGROUND: Vasospastic disorders (acrocyanosis, Raynaud's syndrome, causalgia) can arise from different etiologic factors, but the pathogenesis is always represented by an altered mechanism of vasal motility. Upper dorsal sympathectomy has been demonstrated to be an effective treatment for these disorders by decreasing peripheral resistances. Surgical technique has shown long-lasting results, and it can now be performed by endoscopic approach. METHODS: Our experience with six cases of sympathectomy is illustrated. The indications, thoracoscopic technique, complications and long-term results are evaluated. Four women with Raynaud's syndrome and two men with causalgia were treated in this series. After an accurate preoperative evaluation, the second, third, fourth and fifth thoracic ganglia of the sympathetic chain were identified and excised. RESULTS: All patients experienced relief of symptoms with very limited pain and discomfort. They did not require further medical therapy and are relapse-free at follow-up. CONCLUSIONS: We conclude that thoracoscopic sympathectomy can be considered an effective, safe and simple treatment for selected cases of vasospastic phenomenon. PMID- 9876749 TI - Minimally invasive anterior spinal exposure and release in children with scoliosis. AB - BACKGROUND: The early experience with thoracoscopy in children has involved the diagnosis and treatment of pleural and pulmonary diseases. Recent advances have allowed surgeons to perform more complex procedures through video-assisted thoracoscopic surgery (VATS), potentially decreasing the pain and pulmonary impairment associated with an open thoracotomy. The authors report their initial experience with thoracoscopic assisted anterior spinal exposure and release as part of the treatment for children with spinal deformities. METHODS: A retrospective chart review of five children who underwent VATS for anterior spinal surgery between June 1995 and January 1997 was performed. RESULTS: The ages of the patients ranged from 11 to 16 years with a mean of 13.4 years. All patients had an anterior spinal release with or without fusion and same-day posterior spinal fusion with instrumentation. VATS was successfully completed in all patients without major morbidity and no mortality. The average operative time for the anterior portion of the procedure was 305 minutes, and a mean of 7 disc levels were released. Mean length of chest tube drainage and hospitalization were 6.8 and 8.6 days, respectively. CONCLUSIONS: The objectives of anterior exposure for spinal surgery in children can safely and effectively be accomplished using minimally invasive surgery. PMID- 9876750 TI - Laparoscopic orchidopexy: the treatment of choice for the impalpable undescended testis. AB - OBJECTIVE: Management of the impalpable testis often represents a significant diagnostic and therapeutic challenge for the radiologist and surgeon. The aim of this work is to present the superior value of laparoscopy as a single tool for the diagnosis and treatment of the impalpable testis. METHODS AND PROCEDURES: Thirty-three patients with 43 impalpable testes are included in this study. For each patient, magnetic resonance imaging (MRI) then laparoscopy were performed. Either the testis or blind-ending cord structures are searched for. The testis was either brought down to the scrotum or removed depending on the condition of each patient. RESULTS: MRI detected 16 out of 43 impalpable testes (37.2%). Laparoscopy, on the other hand, detected 18 intra-abdominal testes, 7 inguinal and 16 blind-ending cord structures either above or below the internal inguinal ring. Blind-ending cord structures suggested a vanished testis. Orchidopexy was done for 23 cases, orchidectomy in 18 cases and laparoscopy only in 2 cases. CONCLUSION: Laparoscopy seems to offer a safe and reliable diagnostic and therapeutic option to patients with impalpable testes. Intra-abdominal dissection allows more testes to be brought down to the scrotum. The procedure is best viewed as laparoscopy-assisted, as orchidopexy has to be done in a conventional manner. PMID- 9876751 TI - The fate of retained gallstones following laparoscopic cholecystectomy in a prairie dog model. AB - BACKGROUND AND OBJECTIVES: Reported complications of retained gallstones following laparoscopic cholecystectomy (LC) are increasing. This study was undertaken to evaluate the effects of retained gallstones following LC in a prairie dog model. METHODS: Twenty-seven prairie dogs with diet-induced gallstones were divided into three groups of nine. Group I (control) had LC with removal of stones. Group II had LC followed by return of native stones intra abdominally. Group III had LC followed by return of infected stones (stones dipped in Escherichia coli) intra-abdominally. Animals were euthanized at two months and the character and extent of intra-abdominal adhesions were scored. RESULTS: Adhesions were present in 56% of animals in Group I, 89% in Group II, and 100% in Group III. The character and extent of adhesions in groups II & III were significantly greater than the control group (p < 0.03). Group III exhibited the highest degree of adhesions when compared to control (p < 0.007). Histopathology revealed evidence of micro-abscess formation, foreign body giant cell reaction, and fat necrosis adjacent to retained stones. CONCLUSION: Retained intra-abdominal gallstones, especially if infected, are associated with increased adhesions and inflammatory response in this LC model. Further investigation into the long-term consequences of this entity is warranted. PMID- 9876752 TI - Laparoscopic paraesophageal hernia repair. AB - BACKGROUND AND OBJECTIVE: Paraesophageal hernias are uncommon yet potentially lethal conditions. Their repair has now been facilitated by laparoscopic technology. We present a series of 20 patients with paraesophageal hernias repaired laparoscopically. METHODS: Twenty patients with paraesophageal hernias had laparoscopic repairs. Eighteen patients had primary repair of their hiatal defect. Two required mesh reinforcement. Fifteen patients had a fundoplication procedure performed concomitantly. RESULTS: Long-term follow-up is available on 17 patients. There was no in-hospital morbidity or mortality. Average length of stay was 2.3 days. One patient recurred in the immediate postoperative period. There were no other recurrences. The only death in the series occurred in the oldest patient 18 days postoperatively. He had been discharged from the hospital and died of cardiac failure. No patients have had complications from a paraesophageal hernia postoperatively. CONCLUSION: Laparoscopic repair of paraesophageal hernias is possible. Preoperative work-up should include motility evaluation to assess esophageal peristalsis as the majority of these will need a concomitant anti-reflux procedure. This data helps the surgeon to determine whether or not a complete or partial wrap should be done. Repair of the diaphragmatic defect can be accomplished in the majority of patients without the use of prosthetic material with excellent results. PMID- 9876754 TI - Bowel obstruction in the postoperative period of laparoscopic inguinal hernia repair (TAPP): review of the literature. AB - A case is presented of a male patient that presented with intestinal obstruction in the early postoperative period of a transabdominal preperitoneal inguinal repair (TAPP) that was diagnosed and repaired successfully using laparoscopy. Whenever a total extraperitoneal procedure (TEP) cannot be performed, the peritoneal closure of the TAPP should be done water-tight using interrupted stitches of absorbable monofilament sutures. PMID- 9876753 TI - Post-laparoscopic vomiting in females versus males: comparison of prophylactic antiemetic action of ondansetron versus metoclopramide. AB - BACKGROUND AND OBJECTIVES: The incidence of postoperative vomiting in patients undergoing laparoscopic cholecystectomy is compared in females versus males. The report also compares the prophylactic action of ondansetron versus metoclopramide. METHODS: A total of 85 American Society of Anesthesiologists (ASA) I and II patients were enrolled in the study. Patients were divided into two groups according to sex: Group I 53 females, and Group II 32 males. After anaesthetic induction, subjects received intravenously either 4 mg ondansetron or 10 mg metoclopramide. RESULTS: The incidence of vomiting as well as the frequency of emetic episodes over 24 hours were analyzed in each group using X2 analysis. Data analysis revealed a significantly higher incidence (P < 0.05) of postoperative emesis in females 10:53 (18.9%) as compared to males 0:32 (0%). In the male group, no patient vomited postoperatively, whether prophylactic ondansetron or metoclopramide was used. While the incidence of emesis in the female group was lower (P < 0.05) in the ondansetron group (17.6%) than the metoclopramide group (29.6%). CONCLUSION: These results may indicate prophylactic antiemetic therapy in female patients undergoing laparoscopic cholecystectomy; ondansetron appears to be superior to metoclopramide. PMID- 9876755 TI - Laparoscopic duodenojejunostomy for treatment of superior mesenteric artery syndrome. AB - BACKGROUND AND OBJECTIVES: Superior mesenteric artery (SMA) syndrome is a rare disorder, recognized as weight loss, nausea, vomiting, and post-prandial pain due to compression and partial obstruction of the third portion of the duodenum by the SMA. If conservative treatment fails, then laparotomy with duodenojejunostomy or lysis of the ligament of Treitz is indicated. Recently, laparoscopic division of the retroperitoneal attachments of the duodenum has been described. We report the first case of laparoscopic duodenojejunostomy as the definitive treatment of vascular compression of the duodenum. METHODS: A very thin woman with a diagnosis of SMA syndrome was prepared for surgery after having failed medical therapy. The patient was placed in a supine position, and four laparoscopic ports were required to perform a 5 cm duodenojejunostomy. RESULTS: The patient did well postoperatively. A gastrograffin study revealed no leak with patency of the duodenojejunal anastomosis. She was subsequently discharged home on a regular diet on postoperative day four. CONCLUSION: Laparoscopic duodenojejunostomy is a viable option to treat vascular compression of the duodenum. It provides definitive treatment while preserving the benefits of minimally invasive surgical techniques in the debilitated patient. PMID- 9876756 TI - Tandem subdiaphragmatic and pleural sequelae due to lost gallstones following cholecystectomy. AB - We report two similar thoracoabdominal complications we encountered due to retained gallstones after cholecystectomy. These patients had had an open cholecystectomy after a failed laparoscopic attempt, with spillage of gallbladder debris intraoperatively. They were admitted more than 12 months later with subdiaphragmatic abscesses. Attempted computerized axial tomography (CT) guided drainage of these abscesses resulted in these patients developing pleural fluid collections, which required surgical drainage. The patients underwent exploratory laparotomies, and drainage of the subdiaphragmatic abscesses had revealed gallstones within the abscess cavity. A detailed presentation of these cases, with review of current literature and clinicopathologic issues for discussion are described. PMID- 9876757 TI - Small trocar perforation of the small bowel: a case report. AB - Although laparoscopy is one of the most common surgical procedures done today, bowel perforations can and do occur during the initial insertion of the Veress needle and/or trocar. Recent advances in microlaparoscopy have reduced the morbidity of this complication when encountered. We report a case of small bowel perforation following insertion of a Veress needle with its 2 mm trocar and our minimally invasive intra-operative and postoperative management of the patient. PMID- 9876758 TI - Laparoscopic resection of solitary adrenal metastasis from lung carcinoma: a case report. AB - BACKGROUND AND OBJECTIVES: Patients with adrenal metastases from bronchogenic carcinoma are considered incurable and any surgical treatment is usually excluded. A review of the few cases of adrenalectomy for metastases from lung cancer that have been reported in the literature shows that good results can be achieved in selected patients. We propose a laparoscopic approach to perform the adrenalectomy in these patients. METHODS: A right laparoscopic adrenalectomy for metastasis from lung adenocarcinoma was performed. The right adrenal was resected using the anterior transperitoneal laparoscopic approach. RESULTS: The tumor was resected in total. The operating time was two hours. One year after surgery the patient remains well. CONCLUSIONS: The current indications for laparoscopic adrenalectomy can include the removal of small metastatic adrenal lesions in selected cases. PMID- 9876759 TI - Hand-assisted laparoscopic transhiatal esophagectomy using the dexterity pneumo sleeve. AB - BACKGROUND: As the use of laparoscopic techniques have expanded to more complicated procedures, limitations of laparoscopy have begun to be realized. To help regain the ability to palpate and bluntly dissect tissues, and handle larger organs, surgeons have begun to utilize devices which allow the surgeon's hand to be inserted into the abdomen during laparoscopic procedures. One such devices is the Dexterity Pneumo Sleeve, which was used here to perform transhiatal esophagectomy in two patients. METHODS: Two patients with adenocarcinoma of the esophagus underwent hand-assisted laparoscopic transhiatal esophagectomy. The stomach mobilization was carried out using laparoscopic technique facilitated by the use of the surgeon's hand. Blunt dissection of the esophagus through the hiatus was then carried out. Dissection of the proximal esophagus and creation of a cervical esophagogastric anastomosis was then performed in the neck through a cervical incision. RESULTS: The Pneumo Sleeve proved useful for handling the stomach, as well as, blunt dissection of the esophagus while still maintaining the benefits of laparoscopy, including small incisions, and no postoperative ileus. CONCLUSION: Hand-assisted laparoscopic esophagectomy can be carried out with good, early, postoperative recovery. PMID- 9876760 TI - Splenic retrieval following laparoscopic splenectomy. PMID- 9876761 TI - The making of an instrument: from concept to market. AB - This is an account of the steps one goes through in the development of a new device or instrument. It starts with the conditions that generate the need and then the concept of a new instrument and goes through the process of designing it and protecting it with a patent; it then proceeds through the development of a working prototype and a final refined product. It provides an outline of the steps needed to get the device into the national or international market by selling or licensing it to a company willing to develop it. To be able to demonstrate this process of invention and give real life to the steps involved in the making of an instrument as mentioned above, I describe the circumstances that generated the idea and the development of the Christoudias Tissue Approximator Grasper. The patent is published as issued to demonstrate its different components. PMID- 9876762 TI - Kurt Semm and the fight against skepticism: endoscopic hemostasis, laparoscopic appendectomy, and Semm's impact on the "laparoscopic revolution". AB - In the 1970s, Semm developed thermocoagulation, adapted the Roeder Loop, and further invented extra- and intracoporeal endoscopic knotting to achieve endoscopic hemostasis. His numerous technical inventions, especially the electronic insufflator, allowed more complex operations to be performed laparoscopically. His technique, however, was not quickly adopted by the surgical community. When the first fully laparoscopic appendectomy was carried out by Semm in 1980, a veritable storm broke loose. In the opinion of many prominent surgeons, Semm exaggerated the problem of adhesions, and laparoscopic technique itself was regarded as very dangerous. Misunderstood by medical scientists, Semm displayed an ability to force his ideas through despite skepticism and suspicion. He realized that endoscopic surgery had tremendous potential, and promoted laparoscopic technique not only in his field of gynecology but among general surgeons as well. In 1985, Muhe, of Boblingen, Germany, used Semm's technique to remove the first gallbladder in the world laparoscopically. Three years later when Semm presented a videotape of his laparoscopic appendectomy in Baltimore, he gave impetus to McKernan and Save of Marietta. Georgia, to carry out the first laparoscopic cholecystectomy in the United States. PMID- 9876763 TI - Clinical experience with an optical access trocar in gynecological laparoscopy pelviscopy. PMID- 9876764 TI - Continuous calcium channel blocker infusion in experimentally induced acute pancreatitis: effects on pancreas and liver function. AB - In this study we examined the effects of continuous calcium channel blocker (CCB) infusion on pancreatic duct-ligated acute pancreatitis (AP) in rabbits. Thirty rabbits were used for this study. Animals in group 1 (n = 10), which served as a control group, underwent dummy operations and received 0.5 microliter/h normal saline via the internal jugular vein. Animals in group 2 (n = 10) with artificially-induced pancreatitis received the same dosage of saline in the same manner. Animals in group 3 (n = 10) with artificially-induced pancreatitis received 180 micrograms/kg/h CCB (Verapamil) via the jugular vein starting from just before pancreatic duct ligation. AP histology score, plasma amylase levels and liver function tests were measured after 48 h. Verapamil infusion did not prevent the rise in plasma amylase levels, nor did it prevent pancreatic inflammation and damage. Serum levels of serum glutamate pyruvate transaminase, serum glutamate oxalacetate transaminase and alkaline phosphatase were significantly elevated in group 2 and significant reductions were seen in the Verapamil treated animals (group 3). The findings in this study imply that a continuous 180 micrograms/kg/h dose Verapamil infusion does not ameliorate the pathogenesis of pancreatitis induced by ligation of pancreatic duct but do not rule out a dose-dependent protective effect. Meanwhile, the lowering of liver function test scores should be considered the beneficial effect of CCBs, and this should be investigated in further studies. PMID- 9876765 TI - Improvement of sensitivity in HLA-DRB1 typing by semi-nested PCR-RFLP. AB - A sensitive method of HLA-DRB1 typing was devised using a semi-nested polymerase chain reaction (PCR) followed by a restriction fragment length polymorphism (RFLP) analysis (semi-nested PCR-RFLP method). The first-round amplification (30 cycles) of the semi-nested PCR was performed using DRB generic primer pairs and the second round of PCRs (20 cycles) were performed using DRB1 group-specific primers. The products of the second round PCRs were digested with restriction endonucleases for the typing of HLA-DRB1 alleles. By this method, HLA-DRB1 typing was possible from 10 pg of genomic DNA extracted from lymphocytes and from 0.5 microliter of 1,000 times diluted blood without DNA extraction. HLA-DRB1 alleles could be typed from a 2-mm long bloodstained cotton thread prepared from 10 times diluted blood and from a 2-mm thread of whole blood bloodstains stored at room temperature for 2 years. From the mixture of blood of two individuals with different genotypes, DRB1 alleles of the minor component were detected down to 1/1,000 of the major component. This semi-nested PCR-RFLP method is useful for HLA-DRB1 typing from extremely small amounts of DNA and from mixed samples. PMID- 9876766 TI - Embedding of laboratory wastes in clay or concrete blocks, with special reference to baking osmic acid and cacodylic acid wastes with clay. AB - Liquid laboratory waste containing osmic acid and cacodylic acid was mixed with potter's clay or hydraulic cement. The clay-waste product was kneaded into blocks and baked in a klin (1,200-1,400 degrees C). The cement-waste product was allowed to harden into concrete blocks. Some of the baked clay blocks and concrete blocks were ground, and immersed in 1 N NaOH or 10% HCI solutions for 3-6 months. X-ray microanalysis of the dried samples of these solutions showed that no leakage of osmium and arsenic occurred in the baked clay embedding, and that some leakage of these agents occurred in the concrete embedding. The present study indicates that the baked clay embedding method is useful for safe storage of dangerous laboratory wastes. Additional experiments suggested that glass embedding is also useful for safe storage of laboratory wastes or harmful metals. PMID- 9876767 TI - Overexpression of c-Met/hepatocyte growth factor receptors in human prostatic adenocarcinoma. AB - Hepatocyte growth factor (HGF) and c-met proto-oncogene product (c-Met) have varied biological functions in different tissues and have been implicated in mitogenic, motogenic and morphogenic responses in both organ regeneration and carcinogenesis. Some studies have suggested that the overexpression of c-Met and epidermal growth factor receptor (EGFR) are associated with growth advantage, while transforming growth factor-beta receptor II (TGF beta R II) is associated with growth disadvantage of human prostatic adenocarcinoma. However, it is unclear if the expression of c-Met correlates with the expression of EGFR and TGF beta R II, and with the proliferative status of human prostatic adenocarcinoma. Using immunohistochemical staining with anti-c-Met (C-12), anti-EGFR (NCL-EGFR) and anti-TGF beta R II (L-21) antibodies, we determined the frequency of expression of c-MET, EGFR, and TGF beta R II respectively in a series of 134 radical prostatectomy specimens. We evaluated the relationship between the expression of these receptors and clinicopathological characteristics. Overall, c Met immunostaining was detected in 54 of 134 (40.3%) cases, EGFR in 45 (33.6%) and TGF beta R II in 64 (48.4%). The overexpression of c-Met was significantly more common in poorly differentiated (P < 0.0001) and in the diffusely infiltrated specimens (P < 0.0005). In contrast, TGF beta R II was significantly overexpressed in the well differentiated specimens (P < 0.0001) and associated negatively with c-Met (P < 0.0001). Overall, these data suggest that c-Met/HGF receptor and TGF beta R II overexpression may be involved in the differentiation of human prostatic adenocarcinoma, c-Met with de-differentiation and TGF beta R II with differentiation. PMID- 9876768 TI - Binding specificities of lectins to immobilized glycoproteins and oligosaccharides differ from those of immobilized lectins to oligosaccharides. AB - The carbohydrate-binding specificities of lectins in solution to glycoproteins and neoglycolipids immobilized on a solid phase were analyzed in order to establish a simple, rapid method for structural analysis of the carbohydrate moieties of small amounts of individual glycoproteins blotted on membrane. Eight glycoproteins containing typical O-linked tetrasaccharides or a series of typical N-linked oligosaccharides of the high-man-nose type, hybrid type, and complex type and 6 neoglycoproteins containing mono- or di-saccharides were dot blotted on membranes and the membranes were then reacted with 8 kinds of horseradish peroxidase-conjugated lectins before and after heat treatment. Neoglycolipids containing the glycoprotein-derived oligosaccharides immobilized on a thin layer chromatography plate were also reacted with lectins. The heat treatment of the membrane increased lectin reactivity toward the glycoproteins. The carbohydrate binding behavior of lectins, Phaseolus vulgaris erythroagglutinin, wheat germ agglutinin, and concanavalin A in solution toward glycoproteins and neoglycolipids immobilized on a solid phase differed from that of immobilized lectins toward oligosaccharides in solution. This difference should be noted in lectin detection of specific carbohydrates of individual glycoproteins on membrane. PMID- 9876769 TI - Results of surgical treatment for ossification of the posterior longitudinal ligament of the thoracic spine. AB - Conservative treatment is ineffective for ossification of the posterior longitudinal ligament (OPLL) in the thoracic spine, and surgical treatment is indicated for most cases, while such cases are not often experienced. In the present study, the results of surgical management involving mainly posterior decompression for this disease were evaluated clinically. The study included 9 patients (1 man and 8 women) who underwent surgical treatment for OPLL of the thoracic spine between 1984 and 1993. Laminectomy was performed in 5 patients, and laminectomy plus anterior decompression of the OPLL via the posterior approach based on Otsuka's method was performed in 2 patients. In 1 patient, laminoplasty for OPLL of the cervical spine was combined with laminectomy of the symptomatic lesion in the thoracic spine. One patient underwent anterior decompression and fusion. The results were evaluated using the Japanese Orthopaedic Association score (JOA score) and recovery rate. The postoperative follow-up period ranged from 1 year to 10 years and 3 months (mean, 4 years and 6 months). The mean JOA score was 4.8 before surgery and improved to 7.6 at the final examination. This was a mean recovery rate of 50.1%. Symptoms caused by OPLL in the thoracic spine can be alleviated by posterior decompression where OPLL extends from the upper to the middle thoracic spine or extends from the middle to the lower thoracic spine. It seems, however, that OPLL localized to the middle thoracic spine requires anterior decompression. PMID- 9876770 TI - Can POSSUM, a scoring system for perioperative surgical risk, predict postoperative clinical course? AB - POSSUM, a Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity, is a scoring system which assesses perioperative surgical risks (Copeland GP et al.: Br J Surg, 1991, Vol 78, 356-360). The POSSUM scoring system consists of two categories of assessment to assess the risk of surgery. A 12-factor (age, cardiac status, pulse rate, systolic blood pressure, respiratory status, Glasgow Coma Score, serum concentration of urea, potassium and sodium, hemoglobin concentration, white cell count and findings on electrocardiography) and 4-grade physiological score (PS) were developed. This was combined with a 6-factor (type of surgical procedure, number of procedures, blood loss, peritoneal soiling, presence of malignancy and mode of surgery) and 4 grade operative severity score (OSS). The present paper attempts to validate it retrospectively. Postoperative hospitalization period and duration of antibiotics administration were both significantly correlated with OSS, but not with PS. These results suggest that the POSSUM scoring system is useful for predicting the postoperative clinical course. PMID- 9876772 TI - Mechanisms of rhinitis. AB - Mast cell degranulation leads to the release of histamine and a host of other mediators. However, histamine remains preeminent in proffering the vexatious symptoms of allergic rhinitis including itching, sneezing, dripping, and congestion. The mechanisms of symptom production in rhinitis will be reviewed with an eye toward effects on mucosal resident cells such as epithelium, endothelium, and glands, the role of nociceptive and parasympathetic nerves, and the recruitment and activation of leukocytes in the mucosa. The benefits and mechanisms of action of antihistamines, anticholinergic agents, and glucocorticoids will be discussed. PMID- 9876771 TI - Cytokines and allergic diseases: clinical aspects. AB - In recent years there has been an explosive expansion of knowledge relating to a family of proteins involved in the intercellular communication network of the immune system. These substances, referred to as cytokines, are importantly involved in the highly regulated complex sequence of events of cellular interaction that comprise immune responses. Atopic diseases, which afflict 20-30% of the general population, are now considered to be associated with a set of abnormal genetically regulated immune responses to foreign antigens, i.e., allergens. The atopic individuals is characterized by the excessive production of IgE antibody to allergens after inhalation, ingestion, and surface contact. There are now recognized over 19 major classes of cytokines, which have been organized into the following categories according to their major functional activities: 1) Acute phase reactants, promoting and mediating natural immunity (e.g., IL-1, IL 6, TNF, interferons alpha and beta, and IL-8); 2) Cytokines that mediate cellular growth and differentiation (e.g., IL-7, IL-4, IL-2, IL-5, IL-10, IL-12, IL-13); 3) Cytokines that act as hematopoietic growth factors (IL-3, GMCSF, IL-9, IL-11, stem cell factor); 4) Chemokines (alpha and beta major groups, DTG, RANTES); and 5) Cytokines that exert lymphocyte regulatory activity (EG, IFN-gamma, TGF). Of particular importance to allergic disease is the recent recognition of the regulation of helper immune function by two lineages of T helper cells, i.e., Th1 and Th2, by these cytokines. The Th2 hypothesis of allergy (4) considers atopy as a Th2-driven hypersensitivity reaction to allergens of complex genetic and environmental origins, in which the Th1 lineage, normally driven by IL-2, TNF, and IFN-gamma is deficient, and in which a predominant Th2 response is seen that is driven by IL-4, IL-13, IL-5, and IL-10. This knowledge is finding application in both the diagnosis and therapy of allergic diseases, through the measurement or use of cytokines, which may replace deficient quantities, or the use of anticytokines, which may neutralize elevated quantities of cytokines, events that collectively contribute to the immunologic imbalance characteristic of the allergic state. In the future, the application of cytokines will continue to find clinical application in allergic disease, and it behooves the clinical allergist immunologist to keep abreast of the exciting new developments that are occurring in this field. PMID- 9876773 TI - Nitric oxide in allergic rhinitis and asthma. AB - Nitric oxide is a free radical molecule that can have both homeostatic and proinflammatory roles in the nasal mucosa, sinuses and airway. We have demonstrated elevated levels of nitric oxide in either nasal fluid, serum and spontaneous, or antigen-stimulated mononuclear cells from patients with allergic rhinitis, sinusitis, and asthma. We also showed that various therapeutic agents such as glucocorticoids, theophylline and macrolides can decrease nitric oxide levels in these patients. Thus determination of nasal, serum, mononuclear cell or airway production of nitric oxide may be an important biomarker of inflammation for monitoring newer anti-inflammatory therapy. PMID- 9876774 TI - Airway remodeling: potential contributions of subepithelial fibrosis and airway smooth muscle hypertrophy/hyperplasia to airway narrowing in asthma. AB - Recently, much attention has been focused on the airway structural changes accompanying chronic, severe asthma, and the potential ramifications of these changes for airway function and medical management. Airway remodeling may exaggerate airway narrowing by: (i) thickening of the airway wall internal to the smooth muscle, thereby increasing the luminal obstruction generated by a given degree of smooth muscle shortening; (ii) increasing the amount of smooth muscle, thereby increasing shortening; and/or (iii) reducing the load on the smooth muscle, either by increasing the compliance of the airway wall or by reducing airway-parenchymal interdependence. The possibility also exists that airway remodeling represents a protective mechanism against excessive airway narrowing. The major airway structural changes occurring in asthma are subepithelial protein deposition and increased airway smooth muscle mass (hypertrophy, hyperplasia, or both). Several investigators have found correlations between the magnitudes of subepithelial thickening and smooth muscle hypertrophy/hyperplasia and the severity of airways disease, though interpretation has been made difficult by study differences in patient population, treatment, indices of disease severity, and morphometric technique. Taken together, these data suggest that increases in airway remodeling may contribute significantly to the airflow obstruction observed in patients with asthma. However, data proving a causal relationship between airway remodeling and asthma severity remain elusive. PMID- 9876775 TI - beta-Hexosaminidases: potent mitogens of airway smooth muscle. AB - beta-Hexosaminidases A & B, inflammatory marker enzymes, are mannosyl-rich glycoproteins that are implicated in acute asthma. At physiologically and pathologically relevant concentrations (nM), beta-Hexosaminidases act as potent mitogens of bovine airway smooth muscle cells. This mitogenic action is mediated via 175 kD mannose recognizing receptors that have been isolated from bovine airway smooth muscle cells and human bronchial smooth muscle. There seems to be an involvement of multiple signal transduction pathways in this process and this mitogenic effect is cell density dependent. Moreover, beta-Hexosaminidases are protease resistant. Thus, beta-Hexosaminidases may serve as key inflammatory mediators critical to airway remodeling process. PMID- 9876776 TI - Essential role of T lymphocytes in the development of allergen-driven airway hyperresponsiveness. AB - Asthma now affects more than 15 million Americans and results in significant expenditure of resources. Despite intensive investigation into the pathogenesis of asthma, debate continues over which cells or which mediators are the primary contributors to the disease. Increasingly, asthma is recognized as a chronic, inflammatory disease. T lymphocytes, T-cell derived cytokines, and eosinophils play major roles in the initiation and perpetuation of the inflammatory response. Animal models have enabled us to link directly T cells with eosinophilic inflammation of the airways, providing new insights into pathogenesis and novel opportunities for therapeutic interventions. PMID- 9876777 TI - The presence of atopy does not determine the type of cellular infiltrate in nasal polyps. AB - BACKGROUND AND OBJECTIVES: Any relationship between atopy and nasal polyposis remains to be further studied to determine the contribution of atopy to the pathogenesis of nasal polyps. MATERIALS AND METHOD: We have compared the inflammatory cellular infiltrate in nasal polyp tissue taken during resection from 10 atopic and 11 non-atopic subjects. We have used immunohistochemistry to enumerate the individual inflammatory cell types using monoclonal antibodies against tryptase (AA1) to identify mast cells, the secreted forms of eosinophil cationic protein (EG2) to identify activated eosinophils, neutrophil elastase (NE+) to demonstrate neutrophils, and T cell surface markers (CD3) to identify pan T cells. RESULT: The number of AA1+ and NE+ cells tended to be higher in atopics, but no statistical significance was found (p = 0.06, p = 0.12). Eosinophil numbers (EG2) were abundant in both groups and found to be not different between them (p = 0.65). Some subjects had CD3+ cells with no significant difference between atopic and non-atopic subjects (p = 0.21). Significant correlation was found between NE+ and AA1+ or EG2 cells (r = 0.59, r = 0.63, p < 0.05, respectively). CONCLUSION: These results suggest that the presence of atopy does not determine either the type of extent of cellular infiltration of nasal polyps. PMID- 9876778 TI - Azelastine reduces mediators of inflammation in patients with nasal polyps. AB - Nasal polyps affect approximately 4% of the population in the western world. The etiology of this disease is unknown, although inflammatory mechanisms may play an important role. In preceding studies we and others have shown that besides H1 antagonism, azelastine influences the immigration and activation of inflammatory cells. In this open label study in 16 patients with nasal polyps and perennial mite-allergic rhinitis, the effect of azelastine nasal spray twice daily 0.14 mg to each nostril on recurrence of nasal polyposis after endonasal surgery was evaluated. One patient dropped out after 3 months, unwilling to take further medication. Clinical and laboratory data of 15 patients were recorded over 25 weeks in a total of seven visits. Of these one patient needed nasal budesonide during the 4 weeks between visits 3 and 4. All other patients did not take any steroids before inclusion into the trial or during the 6-month observation period. Concentrations of eosinophil cationic protein (ECP) for eosinophils, myeloperoxidase (MPO) for neutrophils and tryptase for mast cells were determined in nasal secretions before and after eight and 25 weeks of treatment using double antibody radioimmunoassays, because these have been demonstrated to be good inflammatory markers in nasal diseases. Mean concentrations of MPO decreased from 2724 ng/mL to 1610 ng/mL (p = 0.0015) over the entire treatment period. ECP decreased from 458 ng/mL to 264 ng/mL (p = 0.0342). Tryptase decreased from 37.9 ng/mL to 22.4 ng/mL (p = 0.0574). These data were consistent with a significant decrease in clinical symptoms. Thus, azelastine seems to have an inhibitory effect on eosinophil and neutrophil activation in patients with nasal polyps and mite allergy. PMID- 9876779 TI - Paul de Gara, M.D.: President John F. Kennedy's allergist. PMID- 9876780 TI - Asthma among the famous. Supplement. John F. Kennedy (1912-1963), 35th President of the United States. PMID- 9876781 TI - Asthma among the famous. Robert Joffrey (1928-1988), American dancer and choreographer. PMID- 9876782 TI - Asthma among the famous. Ernest N. Morial (1929-1989), American political leader and city mayor. PMID- 9876784 TI - The Omnibus Reconciliation Act of 1990: redefining pharmacists' legal responsibilities. PMID- 9876783 TI - Protecting genetic privacy by permitting employer access only to job-related employee medical information: analysis of a unique Minnesota law. PMID- 9876785 TI - An analysis of genetic discrimination legislation proposed by the 105th congress. PMID- 9876786 TI - Rejecting organs: the organ allocation process and the Americans with Disabilities Act. PMID- 9876787 TI - AIDS & HIV: evidence of actual exposure--Majca v. Beekil. PMID- 9876788 TI - Propofol maintenance to reduce postoperative emesis in thyroidectomy patients: a group sequential comparison with isoflurane/nitrous oxide. AB - The clinical benefit of propofol anaesthesia in the prevention of postoperative nausea and vomiting (PONV) is still being elucidated despite many studies to date. In this study 64 adult female patients scheduled for thyroidectomy received, in a randomized double-blind fashion, propofol with air or isoflurane with nitrous oxide for maintenance of anaesthesia. The primary response variable was the presence or absence of vomiting in the first six hours. A group sequential design was used to allow interim analysis. After 64 patients, the fourth analysis showed that fewer patients receiving propofol vomited or required an anti-emetic during the first six hours (P < 0.05). There was no significant difference detected in the 6 to 24 hour interval. In this group of female patients, total intravenous anaesthesia (TIVA) with propofol is associated with an early reduction in early postoperative vomiting compared with standard inhalational techniques. This reduction in vomiting does not appear to persist beyond the first six hours. PMID- 9876789 TI - Patient-controlled epidural versus intravenous pethidine to supplement epidural bupivacaine after abdominal aortic surgery. AB - In a double-blind, randomized, crossover study of 25 patients after abdominal aortic surgery, we compared patient-controlled analgesia (PCA) with epidural versus intravenous pethidine. All patients received continuous epidural infusions of 0.125% bupivacaine adjusted to maintain appropriate sensory levels. The 48 hour study period commenced 36 to 48 hours after surgery and covered postoperative days 2 and 3. There was a crossover in PCA mode (epidural or intravenous) after 24 hours. Plasma pethidine concentration at the end of each 24 hour period and the total 24 hour pethidine dose did not change significantly between postoperative days 2 and 3. Pethidine plasma concentration was lower after 24 hours epidural than after intravenous PCA [125 (SD 108) ng/ml versus 171 (SD 107) ng/ml, P = 0.03], although pethidine dose did not differ significantly [mean 147 (SD 124) mg/24 h]. Visual analog pain scores (VAS) did not differ significantly between postoperative days 2 and 3, or at rest between epidural and i.v. groups. However, VAS with coughing and with abdominal palpation were lower in the epidural PCA group (P = 0.05, 0.008). With a background epidural infusion of 0.125% bupivacaine, PCA with epidural pethidine provided better pain control than PCA intravenous pethidine and this was achieved at lower plasma pethidine concentrations. PMID- 9876790 TI - The role of preoperative cervical spine X-rays in rheumatoid arthritis. AB - The cervical spine is frequently involved in rheumatoid arthritis and yet there exists no consensus on the need to screen for cervical spine subluxations preoperatively. We reviewed retrospectively 77 patients who underwent 132 operations under general or regional anaesthesia over a 44-month period. We found that while the majority of patients had received preoperative X-ray screening for cervical spine instability, a third of the X-ray examinations done had been inadequate. Many anaesthetists did not repeat cervical spine X-rays if there were previously performed views available. We showed that a complete X-ray examination of the cervical spine should include flexion and extension stress views in addition to frontal views of the odontoid and entire cervical spine. Anterior atlantoaxial subluxation was the most common subluxation encountered in our study population. The detection of cervical spine instability was found to significantly affect anaesthetic management, favouring techniques that avoided unprotected manipulations of the neck under anaesthesia. PMID- 9876791 TI - Management and outcomes of patients with brain trauma in a tertiary referral trauma hospital without neurosurgeons on site. AB - Waikato Hospital is a tertiary hospital of over 700 beds receiving large numbers of trauma patients, but has no neurosurgeon closer than 130 kilometres. Over the 10 years ending July 1997, 831 cases of brain trauma were admitted to the Intensive Care Unit. Of these, 191 died before leaving hospital (overall mortality 23%). Of the 547 who had a Glasgow Coma Score (GCS) < or = 8, 173 died (mortality 32%). Of the children who were < 15 years of age and had a GCS < or = 8, there was a 23% mortality. These mortality rates are acceptable when compared with other reports (average 37%, over 12 adult series). Using brain AIS scores, our mortality figures also compared favorably with those in the literature, and suggest that the quality of brain trauma care is adequate in this non neurosurgical centre with intensive care, backed by CT scanning and general surgeons able to do urgent burr holes. Six percent of the brain trauma patients (approximately five per year), required interhospital transfer for definitive neurosurgical care. PMID- 9876792 TI - The use of antimicrobials in ten Australian and New Zealand intensive care units. The Australian and New Zealand Intensive Care Multicentre Studies Group Investigators. AB - A prospective standardized collection of clinical, microbiological and pharmaceutical information on antibiotic use was conducted in Australia and New Zealand intensive care units (ICUs) involving 481 consecutive critically ill patients who were receiving antibiotics for any reason while in ICU. Patients had a mean SAPS II score of 34.1 +/- 17.8 with an expected mortality of 15.6% (actual mortality 12%). Of these, 292 (60.8%) were admitted to the ICU within 72 hours of surgery. Among such surgical patients, 233 (79.9%) received antibiotics for "surgical prophylaxis" while in ICU (48% of sample population). The second largest group of patients treated with antibiotics in ICU included those with systemic inflammatory response syndrome and clinical suspicion of infection (38%). Antibiotics were prescribed for the treatment of clinically diagnosed infection in 268 patients. Clinical response was apparent in 62.6% and in most (71%) was achieved in the first 72 hours of treatment. The incidence of antimicrobial-related side-effects was 4%, mostly in the form of diarrhoea or rash (75% of all side-effects). The most commonly prescribed antimicrobials were gentamicin (n = 146), ceftriaxone (n = 98), vancomycin (n = 94) and metronidazole (n = 111). Three times daily prescription of aminoglycosides was uncommon (< 1%). Forty-one patients had a documented infection (positive culture) with a gram negative organism. Of these, 17 received therapy with a single antibiotic and 24 received therapy with two antibiotics. Despite similar illness severity, there were six deaths in the former group and only two in the latter. PMID- 9876793 TI - Anaesthesia for transmyocardial laser revascularization--initial experience with seven patients. AB - The introduction of transmyocardial laser revascularization for ischaemic heart disease has brought with it new challenges for anaesthetists. These include acute deterioration of cardiac function, the need for emergency cardiopulmonary bypass and difficulty weaning from haemodynamic support. Recurrent arrhythmias can occur despite amiodarone prophylaxis. We describe our initial experience and the problems we encountered. PMID- 9876794 TI - Improving the evidence base for anaesthesia. MASTER Anaesthesia Trial Study Group. AB - This paper is a brief report of the symposium, "Improving the Evidence Base for Anaesthesia and Intensive Care", organized by the MASTER Anaesthesia Trial Study Group at the Annual Scientific Meeting of the Australian and New Zealand College of Anaesthetists, Newcastle, N.S.W., on Tuesday, May 5, 1998. PMID- 9876795 TI - Multiple cranial nerve palsies complicating retrobulbar eye block. AB - This report details a case of accidental brainstem anaesthesia complicating retrobulbar block for cataract surgery in an 83-year-old man. Prompt diagnosis and treatment resulted in a successful outcome. The many possible presentations of brainstem anaesthesia are described and causative mechanisms are discussed. PMID- 9876796 TI - Disconnect alarm failure in detection of common gas outlet disconnection. AB - This laboratory study was prompted by two paediatric cases where low pressure alarms and capnography failed to detect common gas outlet disconnection when using a T-piece pump ventilator. A carbon dioxide producing model lung was ventilated using the Clare ventilator (a T-piece pump type ventilator) via an Ayres T-piece. The T-piece used has 3 mm diameter fresh gas tubing. Common gas outlet disconnects were carried out using varying fresh gas flows and ventilatory patterns. At fresh gas flows of less than 8 l/min, the low pressure alarm of the Clare ventilator failed to sound following common gas outlet disconnection. Oxygraphy showed a rapid rise in inspired oxygen. End-tidal CO2 rose, whilst inspired CO2 remained at zero. This experiment demonstrates that the Clare ventilator's low pressure alarm detects common gas outlet disconnection poorly when used with an Ayres T-piece with narrow fresh gas tubing. Graphical representation of airway oxygen content has merit as an alerting monitor for common gas outlet disconnection. PMID- 9876797 TI - Difficult pulmonary artery catheterization in a patient with persistent left superior vena cava. AB - A persistent left superior vena cava is an uncommon congenital abnormality. It arises when the left anterior cardinal vein fails to regress during the embryonic period. Although such patients are usually asymptomatic, they may have associated cardiovascular abnormalities. The anaesthetist may encounter difficulty in the insertion of pulmonary artery catheters. Other implications in the management of these patients in the operating theatre or intensive care unit are discussed. PMID- 9876798 TI - Carbon dioxide embolism following diagnostic hysteroscopy. AB - A 50-year-old woman ASA 2 underwent carbon dioxide hysteroscopy under general anaesthesia. Monitoring showed a sudden and rapid fall in end-tidal carbon dioxide followed by oxygen desaturation. She became pulseless and cyanosed. Resuscitation with oxygen, intravenous adrenaline and head-down tilt restored her to haemodynamic stability. Hyperbaric therapy was also administered as air embolism could not be excluded. PMID- 9876799 TI - Fulminant desquamative interstitial pneumonitis. AB - A previously well, 48-year-old female presented with a short history of respiratory distress and fever for which no cause could be found. Open lung biopsy was not contributory. She died within nine days of presentation of hypoxia and multi-organ failure. Post mortem examination revealed the pattern of desquamative interstitial pneumonitis. PMID- 9876800 TI - "Near drowning" due to hydatid disease. AB - Spontaneous rupture of a hydatid cyst is reported in a 19-year-old cattleman. Mechanical ventilation for respiratory failure and a right lower lobectomy were required for flooding of the airway with hydatid cyst fluid. The postoperative course was complicated by the development of acute respiratory distress syndrome. Complete recovery occurred with no recurrence following treatment with albendazole, praziquantel and partial hepatectomy. PMID- 9876801 TI - The anaesthetic management of autistic children. AB - Autistic children are difficult to manage and there are no anaesthesia studies to suggest management strategies. We present five case reports which describe an integrated management program taking into account the special needs of autistic children and their families. We describe a method of early warning and recognition of these patients and the establishment of a database to allow review of our program. We also present a process to minimize the stress and problems inherent in the conventional admission process. Oral ketamine (6 to 7 mg/kg) has proven to be the most reliable preoperative sedative for these patients. PMID- 9876802 TI - Dosage of intrathecal pethidine and body weight. PMID- 9876803 TI - Inhalational induction with sevoflurane in an adult with asthma and tracheal stenosis. PMID- 9876804 TI - Cervical spine instability and the intubating laryngeal mask--a caution. PMID- 9876805 TI - Brodifacoum. PMID- 9876806 TI - Predictive performance of ten equations for estimating creatinine clearance in cardiac patients. Iohexol Cooperative Study Group. AB - OBJECTIVE: The predictive performance of 10 equations used to estimate creatinine clearance (Clcr) was assessed retrospectively from data collected on 420 patients. DESIGN: This study is a retrospective data analysis of information collected on hemodynamically stable patients awaiting coronary angiography during the Iohexol Cooperative Study. SETTING: The Iohexol Cooperative Study was a multicenter study that compared nephrotoxicity of high- and low-osmolar contrast media in patients undergoing coronary angiography. Data used for this analysis were preangiography 24-hour urine collections that were primarily collected in hospitalized patients. PATIENTS: Patients selected from the Iohexol Cooperative Study database for analysis were participants categorized into one or more of six subgroups: elderly (n = 222), hypoalbuminemic (n = 25), chronic renal insufficiency (n = 128), low serum creatinine (n = 115), obese (n = 208), and diabetic (n = 191) who had baseline urine collections of at least 24 hours. OUTCOME MEASURES: Predictive performance was assessed using bias, precision, slopes, and y-intercepts. RESULTS: The Salazar-Corcoran equation was unbiased in the entire group as well as in five of the subgroups. The Cockcroft-Gault equation was unbiased in three of the subgroups. All other equations were biased in predicting Clcr in the entire group as well as in at least four of the subgroups. Precision was generally poor. All slopes were significantly different than one and all y-intercepts were significantly different than zero (p < 0.01). Correlation coefficients were between 0.63 and 0.79 with the exceptions of the low serum creatinine subgroup (r values 0.35-0.64) and the Davis-Chandler equation (r values 0.35-0.71 across groups). CONCLUSIONS: Of the equations studied, Salazar-Corcoran and Cockcroft-Gault appear to be the best for predicting Clcr. PMID- 9876807 TI - Pharmacokinetics of intraperitoneal fluconazole during continuous cycling peritoneal dialysis. AB - OBJECTIVE: To investigate the pharmacokinetic characteristics of intraperitoneal fluconazole in patients undergoing continuous cycling peritoneal dialysis (CCPD). DESIGN: Prospective, nonrandomized, single-dose, open-label study. PARTICIPANTS: Five noninfected volunteer CCPD patients. INTERVENTIONS: Patients received a single dose of intraperitoneal fluconazole 200 mg during their long daytime dwell. Blood samples were collected before and 1, 3, 6, 12 (end of first dwell), 24 (after overnight cycling), 48, 72, 96, and 120 hours after dosing. Used dialysate was collected throughout the study. Unless the patient was anuric, urine was collected for the first 48 hours. MAIN OUTCOME MEASURE: Fluconazole concentrations were assayed by gas-liquid chromatography. Pharmacokinetic parameters were calculated using standard noncompartmental techniques. RESULTS: The bioavailability of intraperitoneal fluconazole was 96% +/- 2% over a 12-hour dwell, absorption half-life was 2.5 +/- 1.2 hours, serum elimination half-life was 71.65 +/- 12.76 hours, and volume of distribution was 0.66 +/- 0.13 L/kg. Peritoneal clearance was 5.96 +/- 0.93 mL/min and proportional to total dialysate volume. Renal clearance was proportional to renal creatinine clearance. CONCLUSIONS: Current treatment guidelines for fungal peritonitis suggest fluconazole 200 mg intraperitoneally every 24 hours. Our data suggest that this dose, administered every 48 hours, is more than sufficient to maintain serum and peritoneal concentrations above the minimum inhibitory concentration for most Candida spp. Other factors, such as residual renal function and dialysis prescription, may also need to be considered. PMID- 9876808 TI - Treatment of Medicaid patients with asthma: comparison with treatment guidelines using disease-based drug utilization review methodology. AB - OBJECTIVE: To compare asthma drug therapy in the Iowa Medicaid population with international treatment guideline recommendations and relate differences to patient outcomes. METHODS: Data on asthma drug therapy and respiratory-related medical services (clinic visits, emergency visits, hospital admissions) were abstracted from prescription claims and diagnostic codes of adult Iowa Medicaid patients with asthma (n = 1029). RESULTS: About two-thirds of the population received a prescription for a short-acting beta 2-agonist during the study period. Patients with greater daily use of short-acting beta 2-agonists had more clinic visits (p = 0.004), likely related to illness severity. Almost one-third (29.3%) of 58 patients receiving a prescription of salmeterol did not receive the recommended prescription of a short-acting beta 2-agonist inhaler in the 6-month period. About one-half of patients who should have been prescribed inhaled corticosteroids according to the international guidelines did not receive such a prescription in the 6-month study period. Increased use of inhaled corticosteroids was associated with fewer (p = 0.04) emergency visits. CONCLUSIONS: Suboptimal asthma treatment in the Iowa Medicaid population, especially in regard to the use of inhaled corticosteroids as preventive therapy, was evident after comparison with treatment guidelines. Increased daily use of inhaled corticosteroids was associated with positive patient outcomes in accordance with guideline recommendations. Educational efforts to improve closer adherence to international guidelines both in prescribing practices and patient compliance with medication for asthma management should be undertaken. PMID- 9876809 TI - Ticlopidine-induced phenytoin toxicity. AB - OBJECTIVE: To report a probable case of ticlopidine-induced phenytoin toxicity. CASE SUMMARY: A 72-year-old white man suddenly developed combative behavior, refused to leave his room, stopped eating, and began falling to the floor 6 weeks after being given ticlopidine. The total phenytoin concentration was measured at 43.6 micrograms/mL; the dosage of phenytoin was decreased and the symptoms later resolved. After ticlopidine was discontinued, the patient was rechallenged with the same dose of phenytoin without symptoms of toxicity. DISCUSSION: Possible mechanisms of the drug interaction are discussed with emphasis on cytochrome P450 metabolism. CONCLUSIONS: Clinicians should be aware of this potentially serious drug interaction and either avoid the phenytoin-ticlopidine combination, or monitor closely for phenytoin toxicity. PMID- 9876810 TI - Potential interaction involving warfarin and ritonavir. AB - OBJECTIVE: To report an unexpected decrease in warfarin effect following the addition of ritonavir to the medication regimen. CASE SUMMARY: A 27-year-old patient with advanced HIV taking warfarin for an inferior vena cava thrombus was started on ritonavir, clarithromycin, and zidovudine. The international normalized ratio (INR) decreased over a period of weeks after the addition of ritonavir, clarithromycin, and zidovudine to the drug therapy regimen. The warfarin dosage was almost doubled in order to maintain a therapeutic INR. Months later, when ritonavir alone was discontinued, the INR rose rapidly and the warfarin dose requirements decreased significantly. DISCUSSION: Potential interactions between warfarin and the protease inhibitors are described in the literature. Ritonavir has been shown to be a potent inhibitor of CYP3A4, an enzyme responsible for warfarin metabolism. Potentiation of warfarin effect and subsequent decrease in the warfarin dosage requirement was anticipated following ritonavir administration; however, the opposite occurred. The mechanism of the potential interaction between warfarin and ritonavir is not known, and may represent a complex, multidrug interaction. The paradoxical decrease in the INR is particularly intriguing. CONCLUSIONS: Frequent, careful monitoring of warfarin is recommended when ritonavir therapy is initiated or discontinued in a patient taking warfarin. The potential for either an increase or decrease in the INR should be anticipated. PMID- 9876811 TI - Successful desensitization to dapsone for Pneumocystis carinii prophylaxis in an HIV-positive patient. AB - OBJECTIVE: To report a case of successful desensitization to dapsone for Pneumocystis carinii pneumonia (PCP) prophylaxis in a patient unable to tolerate trimethoprim/sulfamethoxazole (TMP/SMX) desensitization or dapsone at standard doses. CASE SUMMARY: A 37-year-old HIV-positive African-American man was treated for pneumonia with TMP/SMX and then continued on the drug for PCP prophylaxis. After experiencing a pruritic maculopapular rash with TMP/SMX, both at standard doses and after attempting a desensitization regimen to the drug, he was started on dapsone for PCP prophylaxis. He experienced a rash and fever after taking dapsone at standard PCP prophylactic doses. At this time, an 18-day oral dapsone rechallenge by dose escalation was attempted, and it was well tolerated. CONCLUSIONS: This case suggests that utilization of a dapsone desensitization regimen may permit a viable treatment option in patients previously thought to be intolerant to the agent. More regimens of this type should be attempted and the results published, using both dapsone and TMP/SMX, so that standard desensitization treatment guidelines may eventually be adopted. PMID- 9876812 TI - Syndrome of inappropriate antidiuretic hormone associated with vinorelbine therapy. AB - OBJECTIVE: To describe onset of syndrome of inappropriate antidiuretic hormone (SIADH) associated with vinorelbine therapy for advanced breast cancer. CASE SUMMARY: A 50-year-old white woman with a history of advanced breast cancer refractory to other treatment modalities was receiving vinorelbine. Blood chemistries revealed severely depleted sodium and potassium concentrations from a normal baseline within a 7-day period. A recheck of blood chemistries confirmed hyponatremia. The patient was admitted to the hospital and treated for SIADH. After successful treatment, she was given demeclocycline prophylactically and rechallenged with vinorelbine without recurrence of the syndrome. DISCUSSION: SIADH has been reported as a complication of treatment with vinca alkaloids. To our knowledge, this is the first report of this syndrome related to vinorelbine therapy. CONCLUSIONS: Because of its structural similarity to the other vinca alkaloids, vinorelbine is believed to be responsible for SIADH in our patient. Clinicians should be aware of the possibility that vinorelbine may cause SIADH and possibly hypokalemia. PMID- 9876813 TI - Infection and atherosclerosis--focus on cytomegalovirus and Chlamydia pneumoniae. AB - BACKGROUND AND OBJECTIVE: Numerous studies have reported an association of coronary atherosclerosis and restenosis with certain bacterial and viral infections. This article reviews the pathophysiology of atherosclerosis, the role of infectious agents (i.e, cytomegalovirus and Chlamydia pneumoniae) in atherogenesis, and studies supporting the potential beneficial effects of antibiotics or antiviral agents in the management of atherosclerotic disease. DATA SOURCES: English-language clinical studies, abstracts, and review articles pertaining to infectious agents and coronary atherosclerosis. STUDY SELECTION AND DATA EXTRACTION: Relevant seroepidemiologic and pathologic studies and animal models evaluating the role of cytomegalovirus or C. pneumoniae in coronary atherosclerosis. DATA SYNTHESIS: Studies evaluating the possible role of cytomegalovirus and C. pneumoniae in the pathogenesis of atherosclerosis, as well as studies examining the use of antimicrobial and antiviral agents for reduction of cardiovascular events, are reviewed and critiqued. CONCLUSIONS: Current data do not allow us to determine whether infection is a cause or a cofactor of atherosclerosis. These uncertainties can be resolved by larger scale seroepidemiologic, pathologic, and interventional studies. Such efforts will contribute to identifying populations that are appropriate for particular surveillance or specific interventions, such as antibiotics or antiviral therapy. PMID- 9876814 TI - Ambulatory management of chemotherapy-induced fever and neutropenia in adult cancer patients. AB - OBJECTIVE: To review the literature on the management of low-risk adults with chemotherapy-induced fever and neutropenia (CIFN). Included in the review are methods to identify these patients, management options, and economic impact associated with nontraditional treatment options. DATA SOURCES: A MEDLINE and bibliographic search (January 1966-December 1997) for all English-language studies evaluating the identification and treatment of adult, low-risk CIFN patients was completed. Reference lists from identified articles also served as literature sources. STUDY SELECTION AND DATA EXTRACTION: All human studies identified from the data sources were evaluated. Pertinent information, excluding pediatric studies, was selected and critically evaluated for discussion. DATA SYNTHESIS: Alterations in prominent bacterial isolates in CIFN, newer antibiotic choices, enhanced focus on patient comfort, and cost-containment directives have promoted recent research identifying adult cancer patients with low-risk CIFN. Using this information to select low-risk CIFN patients, several investigators have completed trials using antibiotic therapy applicable to the ambulatory setting. Additionally, some investigators have included the use of an oral outpatient antibiotic regimen. Limited data indicate that this approach is a reasonable treatment option for selected patients. CONCLUSIONS: A subset of adult patients with CIFN are at low risk for serious morbidity and mortality when treated with broad-spectrum antibiotics in the ambulatory setting. Managing these patients with this approach requires close patient selection, intense follow-up, data collection, and ongoing evaluation to determine efficacy and patient safety. Currently, ambulatory treatment with oral antibiotics for CIFN is not considered standard of care. Further studies of larger size designed to confirm low-risk patient characteristics and optimal antibiotic selection are required. PMID- 9876815 TI - Irinotecan: a new antineoplastic agent for the management of colorectal cancer. AB - OBJECTIVE: To review the pharmacologic, pharmacokinetic, therapeutic, and safety aspects of irinotecan, a new antineoplastic agent, and to assess its role in the treatment of colorectal and lung cancer. DATA SOURCES: English-language articles from the MEDLINE database, January 1990-March 1998; Pharmacia & Upjohn Company; published articles and meeting abstracts. STUDY SELECTION: Studies in humans with cancer, clinical case reports, and open clinical studies were reviewed. Efficacy studies were limited to trials with at least 20 evaluable patients. DATA EXTRACTION: Relevant data were extracted from published reports and abstracts. DATA SYNTHESIS: Irinotecan is an effective agent for the treatment of advanced colorectal cancer. It demonstrates significant activity as a first-line agent and in patients with disease that is refractory to fluorouracil-containing regimens. Activity against lung cancer has also been demonstrated. Limited data indicate activity against cancers of the ovary, cervix, stomach, and in non-Hodgkin's lymphomas. Major toxicity consists of myelosuppression and diarrhea. CONCLUSIONS: Irinotecan is a useful addition to the antineoplastic drug family and offers significant efficacy for treatment of patients with fluorouracil-refractory colorectal cancer. PMID- 9876816 TI - Topotecan: a new topoisomerase I inhibiting antineoplastic agent. AB - OBJECTIVE: To review the pharmacologic, pharmacokinetic, therapeutic, and safety aspects of topotecan, a new antineoplastic agent, and to assess its role in the treatment of cancer. DATA SOURCES: MEDLINE database English language only, January 1990-March 1998; SmithKline Beecham Pharmaceuticals; published articles, books, and abstracts. STUDY SELECTION: Studies in humans with cancer, clinical case reports, open clinical trials, and controlled clinical studies. Efficacy studies were limited primarily to trials with at least 20 evaluable patients: DATA EXTRACTION: Relevant data were extracted only from published reports. Data were obtained from studies in both articles and abstracts. Only articles written in English were reviewed. DATA SYNTHESIS: Topotecan is an effective second- or third-line therapy for patients with advanced ovarian cancer and is comparable to ifosfamide, liposomal doxorubicin, and paclitaxel. Activity in combination with other agents and as a first-line agent is yet to be determined. Limited data indicate activity in small-cell lung cancer, cancers of the breast and uterus, and in nonlymphocytic leukemia. The dose-limiting toxicity is myelosuppression. CONCLUSIONS: Topotecan is an effective second-line agent for patients with unresponsive or relapsed cancer of the ovary. It appears to be similar to other active agents in patients with this disease status. Its ultimate role in ovarian cancer and other neoplasms awaits additional evaluation in combination with other agents and as first-line therapy. PMID- 9876817 TI - Toxicity management in patients receiving low-dose aldesleukin therapy. AB - OBJECTIVE: To review the pathophysiology and subsequent treatment options for low dose aldesleukin-induced toxicity when administered via intravenous bolus infusion, continuous intravenous infusion, or subcutaneous injection. BACKGROUND: The adverse events associated with high-dose aldesleukin therapy (600,000 IU per kg i.v. every 8 h for a maximum of 14 doses) are well documented in the literature; however, the adverse event profile of lower doses and alternative administration routes are less well described. An understanding of the adverse event profile associated with these alternative regimens can enhance management of toxicity. DATA SOURCES: English-language clinical studies, abstracts, and review articles pertaining to low-dose intravenous, continuous intravenous infusion, or subcutaneous injection of aldesleukin, as well as aldesleukin induced adverse events. STUDY SELECTION AND DATA EXTRACTION: Relevant studies were selected that assist with understanding the pathophysiology, clinical management, diagnosis, and management of aldesleukin-induced adverse events. CONCLUSIONS: Aldesleukin therapy initiates a cytokine-mediated proinflammatory process resulting in a toxicity profile that is different from traditional nonbiologic chemotherapeutic agents. The frequency and severity of adverse events associated with aldesleukin administration are dependent upon dose, route, and administration schedule. In addition, most adverse reactions are self-limiting. Alleviation of aldesleukin-induced adverse effects can usually be achieved on an outpatient basis with agents such as antiemetics, antipyretics, and topical creams or lotions, as well as nonmedication interventions. Aggressive and proactive management of aldesleukin associated toxicities can help facilitate completion of therapy. PMID- 9876818 TI - Therapy of Candida infections: susceptibility testing, resistance, and therapeutic options. AB - OBJECTIVE: Review the epidemiology of fungal infections, approved susceptibility testing methods, the scope of antifungal resistance, and advances in the treatment of fungal infections. DATA SOURCES: MEDLINE databases (from 1966 to March 1998) were searched for literature pertaining to the epidemiology and management of fungal infections. STUDY SELECTION AND DATA EXTRACTION: Articles were selected to assist in providing the reader an understanding of the epidemiology and management of fungal infections. DATA SYNTHESIS: Fungi have emerged as an important class of pathogens. Even though fungi rank as the fourth most commonly encountered nosocomial bloodstream pathogen, and are associated with the highest mortality of commonly encountered pathogens, only within the past year have methods for conducting and guidelines for interpreting in vitro susceptibility tests been approved. Under the guidance of these standards, we have begun to understand important issues regarding fungi such as the scope and mechanisms of antifungal resistance. Although there has not been a significant addition to our antifungal armamentarium since 1992, advances in antifungal therapy have been realized with the reformulation of available agents and the delineation of the pharmacodynamic characteristics of several antifungals. Additionally, several new agents, including a new class of antifungals, probably will enter into clinical use within the next 5 years. CONCLUSIONS: We have entered an era in which our understanding of fungi is increasing tremendously. Clinicians need to familiarize themselves with the current concepts surrounding the management of fungal infections in order to provide optimal care for their patients. PMID- 9876819 TI - Buspirone use for smoking cessation. AB - The results of buspirone efficacy have been inconsistent and contradictory. The rate of smoking abstinence has been reported to range from 36% to 88% and 16% to 89% in buspirone and placebo treatment groups, respectively. Only one controlled study reported buspirone efficacy in reducing nicotine withdrawal symptoms, although it was based on a small sample population and only 4 weeks of follow-up. The most recent studies have been unable to demonstrate the efficacy of buspirone in smoking cessation or in the relief of withdrawal symptoms. A placebo controlled, randomized trial with a large number of patients, relatively high doses of buspirone (30-60 mg/d), strict abstinence criteria, long-term follow-up, and the inclusion of smokers with general anxiety or anxiety reported in previous quit attempts is needed to further evaluate buspirone efficacy in smoking cessation and the reduction of nicotine withdrawal symptoms. The treatment effects of buspirone could then be specifically tested as a function of alleviating the anxiety component of the smoking withdrawal syndrome. Finally, buspirone may prove to be an alternative in patients unsuccessful with or unable to tolerate transdermal nicotine therapy. How buspirone compares with bupropion therapy for smoking cessation is also unknown. PMID- 9876820 TI - Botulinum toxin type A injection for the treatment of frown lines. AB - Patients who have exaggerated frown lines frequently ask for treatment because others mistake them to be constantly angry or annoyed. Current treatment options (surgery or implants) do not address the underlying cause of these lines, namely the excessive nerve stimulation. The mechanism of action of BTX makes it an ideal agent to target the major cause of these lines. BTX inhibits calcium metabolism in the presynaptic neuron, thereby inhibiting neuromuscular transmission and producing muscle paralysis. The current medical literature indicates that BTX can be used safely and effectively for the cosmetic treatment of frown lines. The procedure can be performed in an ambulatory setting and the use of an EMG instrument may provide better direction for the placement of the drug. However, the benefits are transient and repeated injections are necessary. The adverse effects associated with BTX injections were mild and transient. Currently, there are no safety data on the long-term effects of continuous BTX injections for the treatment of frown lines. However, studies on the long-term use of BTX at doses to treat blepharospasm and Meige's disease have shown no serious adverse effects or production of antibodies to BTX. Muscle biopsies taken from patients who have received numerous doses of BTX for more than 7 years have not shown any signs of atrophy or permanent muscle degeneration. The medical literature supports BTX therapy as an option for the treatment of cosmetic facial frown lines. However, there is not enough information on what patient characteristics are ideal in a candidate to achieve optimal response with BTX. Also, no data are available on the safety and efficacy of continuous injections in the long-term use of BTX for facial frown lines. Therefore, the use of BTX to treat frown lines should be examined carefully against other therapeutic options. PMID- 9876821 TI - Penicillamine in the etiology of bullous pemphigoid. PMID- 9876822 TI - Increase in serum carbamazepine concentrations after acute viral hepatitis. PMID- 9876823 TI - Liver injury with the use of ticlopidine. PMID- 9876824 TI - Hepatic injury associated with losartan. PMID- 9876825 TI - Comment: glucosamine: selecting appropriate study exclusion criteria. PMID- 9876826 TI - Comment: mibefradil, a new class of calcium-channel antagonists. PMID- 9876827 TI - Comment: effect on dissolution from halving methylphenidate extended-release tablets. PMID- 9876828 TI - Correction: the role of the ketogenic diet in children with intractable seizures. PMID- 9876829 TI - Control of avian encephalomyelitis: a historical account. AB - Avian encephalomyelitis control methods were not developed until the 1950s although the disease had been discovered and described over 20 yr earlier. Inability to transmit the infection by other than intracerebral inoculation, lack of suitable immunologic methods, the unknowing use of immune chickens or embryos for experimental studies, and reliance on a highly adapted strain of virus rather than fresh field isolates were the main reasons for a general lack of progress. In the absence of supportive experimental data, at least two commercial breeding organizations turned to the use of a crude chicken brain-propagated virus for vaccination of breeder replacement flocks in the 1950s. This control procedure turned out to be practical and efficacious. Development of suitable embryo infection methods and immunologic tests and the chance finding that antibody-free flocks were essential for experimental studies led to the development of embryo susceptibility tests to identify immune breeder flocks and formed the basis for another commercially applied control program, the testing and selection of only immune flocks for hatching purposes. The application of the new testing methods coupled with a switch from the adapted Van Roekel strain of virus to fresh field isolates for experimentation resulted in a rapid unraveling of the epizootiology and pathogenesis of the disease and also to the development of a safe and effective vaccine that was licensed for administration to breeder replacements in 1962. PMID- 9876830 TI - The early history of infectious bronchitis. PMID- 9876831 TI - Effect of a commercial competitive exclusion culture (Preempt) on mortality and horizontal transmission of Salmonella gallinarum in broiler chickens. AB - A commercial competitive exclusion (CE) culture (PREEMPT) was evaluated for its ability to decrease mortality during the first 10-12 days posthatch resulting from the causative agent of fowl typhoid, Salmonella gallinarum. In the first experiment, chicks provided the CE culture on day of hatch and challenged with 10(5) S. gallinarum on day 3 had a significant decrease in mortality compared with non-CE-treated S. gallinarum-challenged chicks in all four replicates. Mortality for control chicks when averaged across all four replicates was 74% compared with 7.5% for the CE-treated chicks. A second experiment was performed that was designed to measure the efficacy of the CE culture in decreasing the horizontal transmission of this pathogen from infected to uninfected chicks when commingled. Day-of-hatch chicks that were directly infected (seeders) with 10(5) S. gallinarum and provided no CE culture averaged 86% S. gallinarum organ positive and 80% mortality during the first 12 days posthatch across four replicates. Untreated contact (not directly infected) chicks that were commingled with the seeder chicks averaged 84% S. gallinarum organ positive and 54% mortality, and the commingled CE-treated contact chicks (provided CE culture on day of hatch) average 35% S. gallinarum organ positive and 9% mortality during the same time period. These results are of importance to the poultry industry in geographic areas where poultry production is adversely affected by fowl typhoid and indicate that treating newly hatched chicks with this commercial CE culture may be a novel way of decreasing economic losses associated with this highly pathogenic organism. PMID- 9876832 TI - The in vitro propagation of stunting syndrome agent. AB - Stunting syndrome (SS) is a viral enteric disease of turkey poults. The etiologic agent (stunting syndrome agent [SSA]) of this disease has been reported recently. The objective of this study was to develop a method for in vitro propagation of SSA. Primary cells, various continuous cell lines, and embryonated eggs were evaluated. Turkey embryos that were inoculated via the amniotic cavity at 24-25 days of incubation were susceptible to SSA infection. The jejunal maltase activity of SSA-inoculated turkey embryos was significantly (P < or = 0.001) lower than that of control embryos. D-xylose absorption was also altered in SSA infected turkey embryos. The extent of reduction of D-xylose absorption and maltase activity in the infected embryos was nearly identical to that observed when day-old poults were infected with SSA. The intestines from the infected turkey embryos were pale, thin walled, and distended with fluid. Electron microscopic examination of the intestinal fluid and epithelial cell lysate of infected embryos revealed pleomorphic membraned SSA viral particles. SSA that had been serially passaged in turkey embryos retained its ability to induce SS in day old poults. All the primary and continuous cells that were evaluated did not support the replication of SSA on the basis of cytopathic effects, electron microscopy, and turkey embryo inoculation. Inoculation of chicken embryos by various routes failed to support SSA. All routes of inoculation, other than the amniotic route at 24-25 days, failed to support SSA in turkey embryos. The results of the this study indicate that the SSA was successfully propagated in turkey embryos that exhibited alterations in embryo intestinal absorption and digestive enzyme activity similar to poults with SS. Successful propagation of SSA in turkey embryos should prove beneficial for future studies including characterization of SSA, prevention and control strategies, and enteric disease modeling. PMID- 9876833 TI - Production of temperature-sensitive clones of Mycoplasma synoviae for evaluation as live vaccines. AB - An Australian field isolate of Mycoplasma synoviae (MS), 89079/7NS, was exposed to the mutagen N-nitro-N'-methyl-N-nitrosoguanidine. Fifteen clones from the exposed culture were characterized for temperature sensitivity. Four clones labelled B, D, G, and H were temperature sensitive and were further characterized for their ability to colonize chickens and elicit an immune response. Serum antibody responses to MS were detected 3 wk after infection, by eyedrop, in 10 of 10 birds inoculated with 86079/7NS and clones B and G and in 9 of 10 birds inoculated with clone H. No MS antibody response was observed in any bird inoculated with clone D. MS was recovered from the upper trachea of 10 of 10 birds inoculated with clones B, G, and H at 2 wk after infection. No MS was isolated from birds inoculated with clone D. Clone H, designated MS-H, was selected as a potential vaccine candidate. PMID- 9876834 TI - Efficacy of a temperature-sensitive Mycoplasma synoviae live vaccine. AB - A temperature-sensitive clone of Mycoplasma synoviae (MS), MS-H, derived from the chemical mutagenesis of the Australian field isolate 86079/7NS, was investigated for efficacy as a live vaccine. Titers of MS-H vaccine ranged between 1.16 x 10(8) and 8.4 x 10(8) color changing units/ml when incubated at 33 C and were consistently > or = 10(2) lower when incubated at 39.5 C. Laboratory-produced MS H vaccine protected 8 out of 10 specific-pathogen-free Webster white leghorn chickens against a combined experimental challenge of aerosols of the virulent wild MS strain 88064/FP and T-strain infectious bronchitis vaccine administered intratracheally. Laboratory and commercially produced vaccines were compared for efficacy against thoracic air sac challenge, with both achieving similar levels of air sac protection. Air sac lesion incidences of five air sacs with lesions out of 32 and four out of 32 were seen in groups vaccinated with laboratory and commercial vaccines, respectively, compared with 13 out of 20 in nonvaccinated specific-pathogen-free hybrid white leghorn chickens. An attempt to determine the dose response of the commercial vaccine was conducted with 0.5, 2, and 4 times the standard dose. Air sac lesion incidences of 16 air sacs with lesions out of 32 in the 0.5 times, 11 out of 32 in the 2 times, and 1 out of 32 in the 4 times dose groups were observed. PMID- 9876835 TI - Safety of a temperature-sensitive clone of Mycoplasma synoviae as a live vaccine. AB - A temperature-sensitive (ts+) clone derived from the Australian Mycoplasma synoviae (MS) field isolate 86079/7NS was produced by chemical mutagenesis with N methyl-N'-nitro-N-nitrosoguanidine and assessed for safety as a live vaccine. This clone, designated MS-H, was assessed for pathogenicity in three different models with air sac lesions as the criterion. No air sac lesions were observed when MS-H was administered to specific-pathogen-free hybrid white leghorn (HWL) chickens by eyedrop at 10 times the normal dose or directly into the thoracic air sacs or as an aerosol administered to specific-pathogen-free Webster white leghorn chickens with concurrent intratracheal T-strain infectious bronchitis virus (IBV). MS-H did not revert to virulence or lose the ts+ phenotype when passaged through five in vivo and 10 in vitro passages. No adverse effects were seen when HWL chickens were vaccinated concurrently with MS-H and combinations of Mycoplasma gallisepticum ts-11 vaccine, IBV vaccine, and infectious laryngotracheitis virus vaccine. Lateral transmission of MS-H was found to occur when vaccinated HWL chickens were mixed with unvaccinated chickens 2 wk after vaccination. At 1 wk after mixing, one out of two unvaccinated chickens had seroconverted to MS and was culture positive for MS. At 2 wk after mixing, both contact chickens were positive for MS by culture and serology. PMID- 9876836 TI - Field evaluation of the safety and efficacy of a temperature-sensitive Mycoplasma synoviae live vaccine. AB - Mycoplasma synoviae (MS) strain MS-H was used in three separate commercial flocks for large-scale evaluation of the safety and efficacy of the vaccine under commercial conditions. MS-H successfully colonized meat and layer-breeders vaccinated by eyedrop and persisted for up to 55 wk after vaccination. Restriction fragment length polymorphism analysis showed that MS-H was the only strain isolated from two vaccinated flocks. In a third flock, challenge with a wild-type MS occurred, and this strain was isolated from both vaccinated and unvaccinated birds. Vertical transmission of MS-H was investigated by culturing pipped embryos and testing broiler progeny for MS antibody at processing (56 days old). No evidence of vertical transmission was detected. Lateral transmission of MS-H strain from vaccinated to unvaccinated birds occurred in one of the commercial flocks. Forty-one of 50 isolates of MS-H obtained from vaccinated flocks maintained their temperature-sensitive phenotype, but nine isolates showed a nontemperature-sensitive phenotype. PMID- 9876837 TI - Antimicrobial resistance and minimum inhibitory concentrations of bacteria isolated from bustards in the United Arab Emirates. AB - The resistance to 11 antimicrobial agents of bacteria isolated from clinical and postmortem cases of five species of bustards maintained in captivity in the United Arab Emirates (UAE) from 1995 to 1997 was determined by disc sensitivity. Of 226 isolates, 169 (74.8%) were gram-negative bacilli, with the majority identified as Escherichia coli (28.8%) and Pseudomonas aeruginosa (19%). The isolates of E. coli showed a high prevalence of resistance to amoxycillin (69%), tetracycline (66.7%), and sulphamethoxazole (43.3%). Moderate resistance to enrofloxacin was observed in E. coli (30%) and P. aeruginosa (19%) isolates. The minimum inhibitory concentrations (MICs) to seven antimicrobial agents were determined for 133 bacterial isolates from bustards during 1996 to 1997. Enrofloxacin (concentration required to inhibit 50% of the isolates [MIC50] < 0.5 microgram/ml) and gentamicin (MIC50 < or = 1.0 microgram/ml) exhibited good activity against both gram-negative bacilli and staphylococci isolated from bustards. Enrofloxacin (MIC50 < or = 1.5 micrograms/ml), gentamicin (MIC50 < or = 2.0 micrograms/ml), and piperacillin (MIC50 < or = 4.0 micrograms/ml) exhibited good activity against P. aeruginosa isolates. Amoxycillin/clavulanic acid showed good activity (concentration required to inhibit 90% of the isolates [MIC90] < or = 2.0 micrograms/ml) against gram-positive cocci and moderate activity (MIC50 < or = 3.0 micrograms/ml) against E. coli, Salmonella spp., and Klebsiella spp. Amoxycillin exhibited good activity (MIC50 < or = 0.5 microgram/ml) against gram positive organisms and moderate activity (MIC50 < or = 1.0 microgram/ml) against Salmonella spp. These results provide information on the prevalence of resistant bacterial strains and the MICs of organisms derived from bustards in the UAE. PMID- 9876838 TI - Construction and characterization of avian Escherichia coli cya crp mutants. AB - We constructed delta cya delta crp mutants of two avian septicemic Escherichia coli strains and evaluated their attenuation in virulence. The P1 phage was used to transfer cya::Tn10 from an E. coli K-12 strain into virulent avian O78 and O2 E. coli isolates. Tetracycline-resistant transductants were plated on Bochner Maloy Medium, and tetracycline-sensitive colonies were selected, then tested by polymerase chain reaction to confirm that they had deletions of the cya gene. Deletions of crp were created by the same technique in isolates with deletions in cya. The delta cya and delta cya delta crp derivatives had slower growth rates, smaller colonies, and impaired fermentation of carbohydrates compared with their wild parents, and they did not revert. Attenuation of the mutant strains was evaluated by subcutaneous (s.c.) inoculation of day-old chicks and by intratracheal (i.t.) inoculation of 9-day-old chicks previously inoculated intranasally with infectious bronchitis virus. For the wild O78 strain and its delta cya and delta cya delta crp derivatives, the percentages of chicks that died within 6 days of s.c. injection of approximately 5 x 10(7) organisms were 100, 60, and 0, respectively. The corresponding percentages for wild-type O2 and its delta cya and delta cya delta crp mutants were 100, 70, and 20 at a dose of approximately 2 x 10(5) organisms. Following i.t. inoculation, group scores based on pathologic and bacteriologic findings were 51%, 15%, and 9% for wild, delta cya, and delta crp O78 strains (inoculum approximately 2 x 10(7) organisms) and 98%, 31%, and 11%, respectively, for the corresponding O2 strains (inoculum approximately 4 x 10(6) organisms). This study demonstrated reduced virulence and stability of the double mutant, which may useful as a live attenuated vaccine against poultry colibacillosis. PMID- 9876839 TI - Alterations in the lymphocytic and mononuclear phagocytic systems of turkey poults associated with exposure to poult enteritis and mortality syndrome. AB - In vivo and in vitro mononuclear phagocytic system functions, expression of lymphocyte subset cell surface markers in the thymus and bursa of Fabricius, and lymphocyte subset dynamics during the course of poult enteritis and mortality syndrome (PEMS) were examined. PEMS is an acute, transmissible, infectious intestinal disease accompanied by high mortality and morbidity. The etiology of this multifactorial disease remains to be elucidated; however, turkey coronavirus was initially assumed to be one of the primary agents involved. Further investigation demonstrated that turkey coronavirus was not always detectable in poults exhibiting PEMS symptoms, and, thus, PEMS poults began to be identified as positive or negative for turkey coronavirus. In each trial, uninfected hatchmate controls were compared with turkey poults that were contact exposed to PEMS poults at 7 days of age. Following intravenous inoculation, control poults cleared Escherichia coli from their circulation by 60 min, whereas viable E. coli were still present in the circulation of PEMS poults at 60 min postinoculation. Inflammatory response measured by Sephadex-elicited abdominal exudate cell recruitment and the adherence potential of abdominal exudate cells was not significantly different between uninfected and PEMS poults. The percentage of glass-adherent abdominal exudate macrophages was higher in PEMS poults. However, the ability of these macrophages to phagocytize sheep red blood cells and the average number of sheep red blood cells per phagocytic macrophage were both lower compared with uninfected controls. CD4+ expression in thymic tissue of PEMS poults at 9 days postinfection was significantly lower. The CD4+:CD8+ lymphocyte ratio in peripheral blood leukocytes from coronavirus-negative PEMS poults was lower than that from both uninfected and coronavirus-positive PEMS poults at 14 days postinfection. In the spleen, the CD4+:CD8+ lymphocyte ratio was higher in coronavirus-positive PEMS poults as compared with the other treatments. In conclusion, immune system dysfunction in PEMS is associated with impaired mononuclear phagocytic system function and alterations in lymphocyte populations. PMID- 9876840 TI - Adhesion inhibition of Mycoplasma iowae to chicken lymphoma DT40 cells by monoclonal antibodies reacting with a 65-kD polypeptide. AB - Tissue- and cell-specific attachment of mycoplasmas is a key aspect of the host parasite relationship. In this study, monoclonal antibodies (MAbs) recognizing surface membrane polypeptides with molecular masses of 46 kD (p46) and 65 kD (p65), respectively, were examined in a microtiter cell attachment (agglutination) inhibition assay. MAbs MI3, MI6, and MI12 reacting with p65 polypeptide of Mycoplasma iowae inhibited attachment of the organisms to chicken lymphoma (DT 40) cells. One MAb (MI2) that reacted with p65 in immunoblots did not inhibit cell attachment, possibly because of the intrinsic native conformation of the epitope(s) in intact mycoplasmas as opposed to the linear state (sodium dodecyl sulfate denatured) in immunoblots. More pronounced M. iowae adherence inhibition was demonstrated by polyclonal turkey and mouse anti-M. iowae antisera compared with MAbs. Immunogold labelling followed by electron microscopy allowed us to localize the MAb-recognized epitopes on the membrane surface of M. iowae. On the basis of the cell attachment inhibition of M. iowae by specific MAbs (MI3, MI6, and MI12), we propose that the p65 polypeptide plays a role in cytadherence. The ability of polyclonal antisera to inhibit attachment of M. iowae more efficiently than the MAbs suggests that additional epitopes within p65 and/or other proteins are involved in cell attachment. PMID- 9876841 TI - Shedding and colonization of Campylobacter jejuni in broilers from day-of-hatch to slaughter age. AB - Poultry are considered to be the primary reservoirs of Campylobacter jejuni for humans. Campylobacter jejuni can colonize the poultry intestinal tract and its subsequent shedding can result in environmental contamination, resulting in an increased risk of infection for the rest of the flock. At present, there is no information on the daily shedding pattern of C. jejuni in broiler chickens. Thus, the purpose of this study was to determine the daily shedding pattern of C. jejuni in broiler chickens and to correlate intestinal colonization with fecal shedding, which would aid in the development of intervention strategies such as the use of competitive exclusion products at 1 day of age. Twenty-four broiler chicks were orally inoculated with 1.6 x 10(7) colony-forming units of C. jejuni, and the reisolation rate of the organism was determined daily from day 1 to day 43. Fifty percent and 70% of the chicks were shedding C. jejuni within 24 and 48 hr postinoculation, respectively. The group collectively reached a peak excretion on days 13-19 postinoculation. There was a steady decline in fecal shedding after the third week. By market age, on day 43, only 37.5% (9/24) of the birds were shedding C. jejuni in their feces. Throughout the sampling period from days 1 to 43, a cyclic pattern of shedding was observed in individual birds. Individual birds excreted C. jejuni on an average of 25 out of 43 days. The C. jejuni isolate failed to colonize 16.6% (4/24) of the birds. A small percentage of the birds, 12.5% (3/24), were observed to be chronic shedders. Enumeration of C. jejuni in the crop, jejunum, and cecum on day 43 revealed that the cecum was the major colonization site, and 15 out of the 24 birds carried C. jejuni in their intestinal tract. PMID- 9876842 TI - In vitro and in vivo comparisons of valnemulin, tiamulin, tylosin, enrofloxacin, and lincomycin/spectinomycin against Mycoplasma gallisepticum. AB - The minimum inhibitory concentrations (MICs) for valnemulin, tiamulin, enrofloxacin, tylosin, and lincomycin/spectinomycin were determined for a virulent strain of Mycoplasma gallispeticum (MG). At the initial reading, the lowest MICs were seen with valnemulin and tiamulin, followed by tylosin, enrofloxacin, and a relatively high MIC for lincomycin/spectinomycin. At the final reading, at 14 days, a similar pattern was obtained, with valnemulin giving the lowest MIC (< 0.008 mg/ml). The same strain of MG was used to infect groups of 20 2-day-old chicks in two separate experiments. In both, several concentrations of valnemulin and tiamulin and one each of tylosin and enrofloxacin were administered to separate groups in the drinking water. In the second experiment, one group of chicks was given lincomycin/spectinomycin. Each experiment had one infected unmedicated group and an uninfected unmedicated group. Mortality, clinical signs, and gross lesions, in both experiments, were significantly less (P < 0.001) in the uninfected and infected medicated groups (except for the two lowest dosages of valnemulin, lincomycin, and spectinomycin) than in the infected unmedicated groups. Also, the mean body weight gain was greater in the uninfected and infected medicated groups. Among the infected birds, MG was recovered from fewer chicks in the infected medicated groups except for the lowest two dosages of valnemulin. Serologic results were negative for the uninfected groups, and there were fewer positive reactors for the infected medicated groups except for the group treated with lincomycin/spectinomycin. Valnemulin should prove to be a useful addition to the antimicrobials in the control of MG infection in chickens. PMID- 9876843 TI - Molecular characterization of an unassigned Israeli Newcastle disease virus isolate. AB - Detection of Newcastle disease virus (NDV) and avian pathotyping of NDV isolates are extremely important because the appearance of virulent virus has significant economic consequences in terms of vaccination, eradication, and the ability to export poultry products. By using nucleotide and amino acid (aa) homology analysis, we could demonstrate that a NDV broiler isolate is a velogenic virus. This analysis was done after mean death time and intracerebral pathogenicity index tests gave inconsistent results. By establishing a nucleotide sequence dendrogram, we found that the disputed Ber-Tuvia was clearly in the same group as the known Herev-Laet, a velogenic isolate. The difference between Ber-Tuvia 92 and the Herev-Laet velogenic isolate was 6% as opposed to > 16% of the meso- and lentogenic isolates. The Ber-Tuvia isolate contains the Arg/Arg and Lys/Arg aa at positions 112, 113 and 115, 116, respectively, in the fusion protein cleavage aa sequence, which is typical for virulent NDV isolates. PMID- 9876844 TI - Bacterin-induced protection of turkeys against fowl cholera following infection with Bordetella avium. AB - Groups of Beltsville small white turkeys, passively immunized and not passively immunized against Bordetella avium, were challenged with live B. avium at 2 days of age. Birds not passively immunized developed severe bordetellosis with early onset, whereas passively immunized birds developed mild bordetellosis with late onset. Following convalescence, birds with and without exposure to B. avium were vaccinated against fowl cholera with a water-in-oil bacterin. The birds were given a homologous challenge with serotype A: 3 Pasteurella multocida. Although no difference in protection against fowl cholera was seen between vaccinated birds that were previously infected with B. avium and those that were not, survivability was better in birds given two doses rather than 1 dose of bacterin. PMID- 9876845 TI - Evaluation of antibody responses of emus (Dromaius novaehollandiae) to avian influenza virus infection. AB - Emu antibody responses to avian influenza virus (AIV) infection were evaluated by the competitive enzyme-linked immunosorbent assay (C-ELISA), agar gel immunodiffusion (AGID) and hemagglutination inhibition (HI) tests. All birds infected with AIV H5N1, H5N3, or H7N7 developed antinucleoprotein (NP) antibodies as early as 7 days postinfection as detected by the C-ELISA. The responses lasted 49 days for the emus receiving H5N3 and at least 56 days for emus receiving the other two viruses. By evaluating 50 emu field serum samples, the C-ELISA was found more sensitive than the AGID test for the detection of anti-NP antibodies. This study indicates that emus experimentally infected with AIV developed antibody responses that can be detected by C-ELISA, AGID, and HI tests. The results from this and our previous studies demonstrate the use of the C-ELISA as a substitute for the AGID test in a routine serodiagnostic screening for detection of antibodies to AIV infection in multiple avian species. PMID- 9876846 TI - Sequence analysis of the VP2 hypervariable region of nine infectious bursal disease virus isolates from mainland China. AB - The VP2 hypervariable region of infectious bursal disease virus (IBDV) from nine Mainland Chinese strains was amplified by reverse transcriptase/nested polymerase chain reaction and cloned into pGEM-T vector. The nine isolates, which were from the center (HN3), the north (Bj-1, B2/28, HD96), the east (JS-18 and AH-2), the northeast (D11-2, C4-2), and the west (Ts) of China, were sequenced and compared with each other and with six reference IBDV sequences. Clustering analysis separated the nine isolate into two groups. The six virulent isolates, propagated in bursae, formed the first group. They revealed only one to three amino acid changes from the very virulent (vv) European and Japanese isolates, suggesting that they might have the same origin as European and Japanese vvIBDV strains. On the basis of their distinct geographic origins, extensive dissemination of vvIBDV in China was indicated. (The other three chicken embryo fibroblast cell cultured isolates with mild pathogenicity were placed in the second group.) Their sequences correlated closely with those of the culture-adapted strains (Cu-1 (4) and Cj-801). None of the nine isolates showed very close sequence relationship with the antigenic variant strains from the USA. Although antigenic variants have been reported in China, the reverse transcriptase/polymerase chain reaction restriction endonuclease analyses of the nine viruses tested herein were not similar to any U.S.A. variant strains on the basis of computer software analysis. Our results and conclusions agree with a previous molecular study of IBDV isolates from the south of China. PMID- 9876847 TI - Early pulmonary lesions in turkeys produced by nonviable Aspergillus fumigatus and/or Pasteurella multocida lipopolysaccharide. AB - This study assessed the potential of lipopolysaccharide (LPS) purified from Pasteurella multocida to cause pulmonary pathology or exacerbate lesions produced by gamma-irradiated nonviable Aspergillus fumigatus conidia when administered via the intra-air sac route in turkeys. LPS provoked suppurative airsacculitis, pleuritis, and pneumonia. Nonviable conidia produced airsacculitis and transient pneumonitis but did not elicit multinucleate giant cells, which are a feature of the inflammatory process in A. fumigatus infection. LPS in combination with A. fumigatus conidia resulted in accelerated pulmonary inflammation and apparently delayed clearance of conidia from pulmonary tissues. This study presents a model of aseptic airsacculitis and pneumonia with clinical relevance. PMID- 9876848 TI - Response of chickens carrying germline insert ALVA11 to challenge with a field strain of subgroup A avian leukosis virus. AB - The ALVA11 germline insert in chickens is a defective subgroup A avian leukosis virus (ALV) proviral insert that expresses a low-to-moderate level of subgroup A ALV envelope glycoprotein. Chicks carrying or lacking ALVA11 were evaluated for response to challenge by RPL-42, a pathogenic field strain of subgroup A ALV, by either exposure to chicks shedding RPL-42 or direct injection with various doses of RPL-42. Chicks carrying ALVA11 were significantly more resistant, as measured by infectious virus and viral antibody status, to horizontal infection and direct injection of RPL-42 than chicks lacking ALVA11. PMID- 9876849 TI - Susceptibility of convalescent turkeys to pulmonary aspergillosis. AB - Pulmonary lesions resulting from Aspergillus fumigatus inoculation were assessed in convalescent turkeys and compared with those in previously noninoculated (control) turkeys. In addition, lesions observed in small Beltsville white (SBW) turkeys were compared with those in broad-breasted white (BBW) turkeys challenged with the same inoculum. Turkeys were challenged by unilateral posterior thoracic air sac (PTAS) inoculation, rechallenged via the contralateral air sac after 5 wk, and then necropsied 1 wk later. Pulmonary lesions induced by the initial challenge had resolved in 6 of 10 SBW and 9 of 10 BBW turkeys. However, convalescence did not protect against pulmonary aspergillosis subsequent to rechallenge; 10 of 10 SBW and 9 of 10 BBW developed granulomatous pulmonary lesions on the side of reexposure. A greater proportion of control SBW turkeys developed pneumonia and airsacculitis following challenge as compared with the BBW breed. Lesions were limited to the lower respiratory tract in all turkeys and were confined to the ipsilateral lung and PTAS in the singly inoculated control turkeys. This study demonstrates that convalescence from pulmonary aspergillosis does not confer protection against rechallenge but may, instead, decrease resistance to subsequent infection. PMID- 9876850 TI - Comparison of an antigen-capture enzyme immunoassay with virus isolation for avian influenza from field samples. AB - The standard tests used to detect avian influenza (AI) viral infection include virus isolation from tissues of the infected birds and the detection of AI antibody in blood or egg yolk. A new application of an existing human test to rapidly detect the presence of any influenza A virus is now possible. A commercially available antigen-capture enzyme immunoassay (AC-EIA), developed for the detection of influenza A in humans was tested for relative sensitivity and specificity and for speed of use in diagnosing nonpathogenic H7N2 AI in naturally infected poultry. During the recent nonpathogenic H7N2 AI epornitic, the AC-EIA was used for rapid diagnosis and quarantine decisions. Between February and August 1997, 1524 samples from 295 commercial layer, pullet, and broiler flocks were submitted to the Laboratory of Avian Medicine and Pathology, New Bolton Center, for AI virus isolation and testing by AC-EIA. The relative specificity of the AC-EIA was 100% and the relative sensitivity was 79%. We believe that the AC EIA will be a useful adjunct to standard AI diagnostic tests. PMID- 9876851 TI - Scanning electron microscopic view of auditory tonsil in turkey. AB - The auditory tonsil of the turkey was studied by scanning electron microscopy. It is a sizable mass of lymphatic tissue dorsal to the infundibular opening in the auditory tube. The infundibular opening is in the roof of the turkey's mouth, caudal to the nasal cleft, and is the external orifice of the auditory tube that connects with the air spaces of the pneumonic bones in the head. Folds of the auditory tonsil that project toward or into the infundibular opening contained numerous lymphatic nodules in their lamina propria. The epithelial surface of the folds was covered with ciliated and nonciliated columnar cells, goblet cells with microvilli, and ductal openings of mucous glands located in the area. The lymphoid nodules were either oval or round and consisted of a thick reticular network that contained lymphocytes, fibroblasts, and erythrocytes. From some of the lymphatic nodules, there were sinusoids that contained lymphocytes and extended to the epithelial surface, whereas from others there was a lymphocytic infiltration of the surrounding lamina propria. PMID- 9876852 TI - Effect of feed withdrawal on Campylobacter in the crops of market-age broiler chickens. AB - The presence of Campylobacter and Salmonella on poultry meat products remains a significant public health concern. Previous research has indicated that feed withdrawal may significantly increase Salmonella contamination of broiler crops and that crop contents may serve as an important source of Salmonella carcass contamination at commercial processing. The present study evaluated the effect of preslaughter feed withdrawal on the incidence of Campylobacter isolation in crops of market-age commercial broiler chickens prior to capture and transport to the processing plant. The incidence of Campylobacter isolation from the crop was determined immediately before and after feed withdrawal in 40 7-wk-old broiler chickens obtained from each of nine separate broiler houses. Ceca were collected from broilers in six of the same flocks for comparison with the crop samples. Feed withdrawal caused a significant (P < 0.025) increase in Campylobacter positive crop samples in seven of the nine houses sampled. Furthermore, the total number of Campylobacter-positive crops increased significantly (P < 0.001) from 90/360 (25%) before feed removal to 224/359 (62.4%) after the feed withdrawal period. Alternatively, feed withdrawal did not significantly alter the Campylobacter isolation frequency from ceca. Similar to our previous studies with Salmonella, the present results suggest that preharvest feed withdrawal increases the frequency of Campylobacter crop contamination and, thus, provides a source of Campylobacter contamination of carcasses at commercial processing. PMID- 9876853 TI - Application of flagella-based immunoassays for serologic detection of Salmonella pullorum infection in chickens. AB - Occasional Salmonella pullorum outbreaks still occur in commercial poultry flocks despite widespread serologic testing, raising concerns about the efficacy of standard agglutination-based serologic assays for detecting infections with current field strains. The recent observation that many S. pullorum strains are recognized by antibodies to certain Salmonella flagellar proteins suggests that similar antibodies might be elicited in infected birds. In the present study, flagella-based enzyme immunoassays were developed and applied to detect antibodies in hens experimentally infected with recent S. pullorum isolates. Antibodies were detected in 29%-44% of serum samples from orally inoculated hens by two flagella-based immunoassays. Similar frequencies of positive results (33% 41%) were obtained by applying two standard agglutination-based tests for antibodies to S. pullorum. Antibodies to flagellar antigens, if also present in naturally infected chickens, could provide an alternative method for detecting S. pullorum infection in poultry flocks. PMID- 9876854 TI - A simple and fast way to count heterophils in chemotaxis. AB - Heterophils were labeled with fluorescein isothiocyanate and then placed in a modified Boyden chamber to determine their response to chemoattractants. After incubation, the cells that had migrated into the membrane were examined under an epifluorescent microscope, and the image was captured by a charge-coupled digital camera and a frame grabber and saved. The saved images were then counted by NIH Image (a public domain software). These modifications greatly speed up the counting of cells and relieve the eyestrain suffered among workers in the chemotaxis field. PMID- 9876855 TI - Serologic evidence of pneumovirus in Chile. AB - Chicken and turkey serum samples were obtained and tested in an enzyme-linked immunosorbent assay to establish the presence or absence of antibodies to avian pneumovirus in the commercial poultry population of Chile. A total of 600 serum samples representing 15 different poultry farms of the central region of Chile were considered. These samples were obtained from 52 different flocks and included 220 turkey and 380 chicken sera. The results showed three flocks of market turkeys belonging to the same farm and three flocks of laying chickens of three different farms to be positive to antibody against pneumovirus. These results provide serologic evidence for the presence of pneumovirus in Chilean poultry. PMID- 9876856 TI - Simultaneous occurrence of Salmonella arizonae in a sulfur crested cockatoo (Cacatua galerita galerita) and iguanas. AB - A case of fatal hepatitis in a captive sulfur crested cockatoo (Cacatua galerita galerita) in which Salmonella arizonae was microbiologically and immunohistochemically detected is described. The death of the cockatoo was closely related to the arrival of a group of 10 green iguanas (Iguana iguana) at a pet shop, and no previous clinical signs were observed in the cockatoo. The most important lesion observed at necropsy of the cockatoo was a multifocal necrotic hepatitis. Salmonella arizonae was isolated from the liver of the cockatoo and was detected immunohistochemically mainly around the edges of necrotic foci. Four iguanas died 3 days later showing a severe enteritis, and Salmonella arizonae was isolated from these lesions. The importance of quarantine and, because of pathogens such as Salmonella, the need to house reptiles at a distance from avian species, mainly psittacids, are reinforced. This is the first report of Salmonella arizonae infection in a cockatoo. PMID- 9876857 TI - Pulmonary sarcomas in a young chicken. AB - Both lungs of a 5-wk-old broiler chicken contained miliary, round, white tumors. Microscopically, these masses were composed of compactly arranged cells with round to oval nuclei and indistinct cytoplasmic borders. Immunohistochemically, tumor cells were positive for vimentin but negative for cytokeratin, S-100 protein, neuron-specific enolase, smooth muscle actin, and muscle-specific actin. Ultrastructurally, tumor cells were separated by collagen bands and some contained collagen fibrils. These results suggest that the primitive mesenchymal cells composing these sarcomas may have originated from precursors of interstitial cells or fibroblasts of the interparabronchial or interatrial septa. DNA was extracted from paraffin-embedded tissues and probed for sequences specific for Marek's disease virus, reticuloendotheliosis viruses, exogenous avian leukosis viruses, and subgroup J avian leukosis viruses. Only sequences specific for subgroup J avian leukosis viruses were identified in the DNA extracted from tumors and surrounding lung. PMID- 9876858 TI - Isolation of avian serotype 3 paramyxoviruses from imported caged birds in Israel. AB - Ten avian serotype 3 paramyxoviruses were isolated for the first time in Israel from passerine and psittacine imported caged birds. The birds were submitted for investigation of an illness characterized by nonspecific signs of weakness, anorexia, vomiting, and sneezing. In addition, only the parakeets developed specific neurologic signs. In bacteriologic and pathologic investigation, cachexia and diarrhea were observed in both groups of birds. In psittacines, considerable alterations were observed in lungs, liver, and spleen. Some nonviral pathogens were occasionally isolated. The isolates appeared to belong to serotype 3b avian paramyxovirus (APMV), the prototype strain of which is APMV 3b/parakeet/Netherlands/449/75. The isolation of APMV-3 viruses from imported caged birds may represent a way of introduction of these viruses into the country. PMID- 9876859 TI - Microsporidiosis in a young ostrich (Struthio camelus). AB - Microsporidia are obligate, intracellular, protozoan parasites of a wide variety of vertebrates and invertebrates. Confirmed reports of microsporidial infection of avian species are few (lovebirds, a parrot, and a group of budgerigar chicks). At slaughter, a 14-mo-old ostrich was found to have small intestinal serosal hemorrhages during postmortem inspection. Histologic examination of the small intestine revealed a chronic lymphoplasmacytic to purulent enteritis with mucosal hyperplasia, muscular hypertrophy, and numerous microsporidia that were located within the superficial enterocytes and the lamina propria. Microsporidia have a ubiquitous distribution in nature and are suspected as possible zoonotic agents. PMID- 9876860 TI - Educating families and caretakers of traumatically brain injured patients in the new health care environment: a three phase model and bibliography. AB - Shorter inpatient stays for traumatically brain injured patients frequently allow less time for comprehensive education of families and caretakers. What is needed to compensate for the shortened duration of brain injury rehabilitation in many settings is a family educational programme that (1) satisfies the family's acute information needs as they arise, and (2) standardizes the delivery of information so that certain knowledge criteria are met. The following article proposes a three phase model of family and caretaker education with an accompanying bibliographical index. PMID- 9876861 TI - A randomized trial of two treatments for mild traumatic brain injury. AB - The purpose of this study was to compare an education-oriented single session treatment (SS) for mild traumatic brain injury (MTBI) to a more extensive assessment, education, and treatment-as-needed intervention (TAN). Participants were 111 adults with MTBI who were recruited from consecutive admissions to two hospital emergency wards. They were randomly assigned to either the SS or TAN modality. The groups did not differ on any demographic, injury-related, or questionnaire variable when first seen within 3 weeks of injury. The groups generally improved a similar amount and did not differ from each other on any symptom-related, functional, or vocational variables 3-4 months after their baseline session. Patient satisfaction ratings with services provided were also similar for the two groups. Brief educational intervention given soon after MTBI appears to be adequate for most MTBI survivors. PMID- 9876862 TI - Neuropsychological testing and functional outcome for individuals with traumatic brain injury. AB - The relationship between performance on neuropsychological measures and the vocational and independent living functioning of individuals with traumatic brain injury was examined. The Wechsler Adult Intelligence Scale-Revised (WAIS-R) IQ and Stroop Color and Word Test scores differentiated individuals who required no assistance with activities of daily living from those requiring some level of assistance. Only the Stroop Color and Word Test scores differentiated individuals who were competitively employed or engaged in degree-oriented education from those who were unemployed or in sheltered or supported employment. Wechsler Memory Scale-Revised (WMS-R) scores did not differentiate these groups. PMID- 9876863 TI - Validity of a cognitive computerized assessment system in brain-injured patients. AB - The primary objective of the study was to investigate the criterion validity of the Cognitive Drug Research (CDR) computerized system in assessing cognitive functioning among persons with brain injury or other organic illness. Understanding the cognitive effects of pharmacotherapy is important in improving long-term rehabilitation outcomes in persons with organic disorders. The CDR system evaluates cognitive skills of attention, short-term or working memory, long-term memory, and visuomotor and motor functioning. For validity testing, correlation coefficients were obtained from an analysis of CDR scores to IQ and Folstein Mini-Mental State Exam (MMSE) scores. Subtests of the CDR system that correlated with the MMSE were choice reaction time (R = -0.542, p = 0.04), spatial working memory (R = 0.938, p = 0.01), and word recognition (R = -0.949, p = 0.01). These tests primarily measured attention and working memory. Correlations between the CDR and IQ scores were not evident. In summary, the data suggest the CDR assessment system has high patient acceptability by persons with traumatic brain injury, and is a useful tool in assessing certain domains of cognition, specifically short-term memory, discrimination, and identification skills. PMID- 9876864 TI - Caregiver burden at 1 year following severe traumatic brain injury. AB - Sixty-nine primary caregivers of adults with a severe traumatic brain injury (TBI) were assessed at 1-year post-injury. Caregivers completed questionnaires on the physical, cognitive, emotional, behavioural, and social functioning of the person with TBI. Caregiver objective burden, psychosocial functioning, and subjective burden were also assessed. Clinically significant levels of anxiety and depression were evident in over a third of the caregivers. Similarly, a quarter of the caregivers reported poor social adjustment. There was no consistent relationship between the prevalence of various types of objective burden and the level of subjective distress that resulted from these changes. The person with TBI's emotional difficulties, in particular their anger, apathy, and dependency, caused the greatest distress for caregivers. With regard to the impact that caregiving had on their own lives, caregivers were most distressed by the loss of personal free time. Results from a regression analysis indicated that the person with TBI's physical impairment, number of behavioural problems, and social isolation were the strongest predictors of caregiver burden. The impact that caring for a person with severe TBI can have on the extended family unit is discussed. PMID- 9876865 TI - A new protocol for training severely impaired patients in the usage of memory journals. AB - Although memory disorders are frequently seen in survivors of brain injuries, remediation of patients with severe memory disorders is still relatively neglected in clinical practice due to pessimism by clinicians regarding the efficacy of such remediation. With respect to memory journal training, a further reason for this neglect is likely the lack of readily accessible protocols that therapists can use to work with these patients. The purpose of the present paper is to describe a new memory journal training protocol that was developed to teach brain injury survivors how to compensate for severe memory impairments by using three strengths of most brain injury survivors: immediate attention, procedural memory, and old learning. A training protocol is described that involves baseline testing and a needs assessment, five levels of training exercises, and discharge probe testing. It is concluded that patients with severely impaired memory abilities can learn the present system when care is taken to individualize journals, conduct proper needs assessments, and provide structured training exercises geared to the strengths of memory impaired persons. PMID- 9876866 TI - Detection of active oxygen species in biological systems. AB - 1. Cypridina luciferin analogues, 2-methyl-6-(p-methoxyphenyl)-3,7- dihydroimidazo[1,2-a]pyrazin-3-one (MCLD) and 2-methyl-6-phenyl-3,7 dihydroimidazo[1,2-a]pyrazin-3-one(CLA ), react with O2- or 1O2 to emit light in visible region. Such chemiluminescences were used for the detection of O2- or 1O2 in activated leukocyte systems and myeloperoxidase (granulocyte-extract) + Br- + H2O2 systems in vitro. 2. The mechanisms of MCLA (CLA)-dependent luminescence is described in detail. Superoxide generated from sinusoidal cells in acute ethanol intoxication of rats was detected by MCLA-dependent luminescence from the surface of perfused rat liver (organ luminescence). 3. Furthermore, with alive animals, O2- generated in the lung of rats with necrotized pancreatitis and that in the stomach of rats after ischemia/reperfusion were detected by their organ luminescences. PMID- 9876867 TI - Usual and unusual methods for detection of lipid peroxides as indicators of tissue injury in cerebral ischemia: what is appropriate and useful? AB - 1. Free radical-dependent lipid peroxidation processes have long been thought to contribute to brain damage following stroke or cerebral ischemia/reperfusion. 2. The preponderance of evidence for this belief has been derived indirectly, through diminution of tissue injury indices (e.g., brain infarct volume) facilitated by application of free radical scavenger substances. 3. Direct, unequivocal evidence for lipid peroxidation in terms of classical assays (detection of conjugated diene absorbance or thiobarbituric acid-reactive substances) is considerably less common, and its validity can be questioned. 4. Correlations of treatment-induced diminishment of brain injury indices with reductions in lipid peroxidation level are rarer still. 5. Reasons underlying the disparity between the belief that lipid peroxidation contributes to ischemic brain injury and direct evidence for this contribution (at least acutely) are proposed, along with evidence that new methods are being developed which should provide the basis for obtaining a definitive answer. PMID- 9876868 TI - Lipid peroxides in the free radical pathophysiology of brain diseases. AB - 1. Polyunsaturated fatty acids are essential for normal cell membrane functioning because many membrane properties, such as fluidity and permeability, are closely related to the presence of unsaturated and polyunsaturated side chains. Lipid peroxidation results in loss of membrane polyunsaturated fatty acids and oxidized phospholipids as polar species contributing to increased membrane rigidity. 2. Polyunsaturated fatty acids are released from membrane phospholipids by a number of enzymic mechanisms involving the receptor-mediated stimulation of phospholipase A2 and phospholipase C/diacylglycerol lipase pathways. 3. The overstimulation of excitatory amino acid (EAA) receptors stimulates the activities of lipases and phospholipases, and this stimulation produces changes in membrane phospholipid composition, permeability, and fluidity, thus decreasing the integrity of plasma membranes. 4. Alterations in properties of plasma membranes may be responsible for the degeneration of neurons seen in neurodegenerative diseases. Two major processes may be involved in neuronal injury caused by the overstimulation of EAA receptors. One is a large Ca2+ influx and the other is an accumulation of free radicals and lipid peroxides as a result of neural membrane phospholipid degradation. It is suggested that calcium and free radicals act in concert to induce neuronal injury in acute trauma (ischemia and spinal cord injury) and in neurodegenerative diseases. PMID- 9876869 TI - Role of superoxide dismutase in ischemic brain injury: a study using SOD-1 transgenic mice. AB - 1. Nitric oxide radicals (NO) play an important role in the pathophysiology of focal cerebral ischemia. 2. Vascular NO can reduce ischemic brain injury by increasing CBF, whereas neuronal NO may mediate neurotoxicity following brain ischemia, mainly by its reaction with superoxide to generate peroxynitrite. 3. These findings could contribute to a strategy for the treatment of cerebral ischemia. PMID- 9876871 TI - Molecular analyses of the Cu/Zn superoxide dismutase gene in patients with familial amyotrophic lateral sclerosis (ALS) in Japan. AB - 1. Amyotrophic lateral sclerosis (ALS) is a degenerative disorder characterized by selective damage to the neural system that mediates voluntary movement. Although the pathophysiologic process of ALS remains unknown, about 5 to 10% of cases are familial. According to genetic linkage studies, the familial ALS (FALS) gene has been mapped on chromosome 21 in some families and recent work identified some different missense mutations in the Cu/Zn superoxide dismutase gene in FALS families. 2. We recently identified five mutations in six FALS families. The mutations identified in our FALS families are H46R, L84V, I104F, S134N, and V148I. The H46R mutation that locates in the active site of Cu/Zn SOD gene is associated with two Japanese families with very slow progression of ALS. On the other hand, the L84V mutation associated with a rapidly progressive loss of motor function with predominant lower motor neuron manifestations. 3. In the family with the V148I, the phenotype of the patient varied very much among the affected members. One case had weakness of the lower extremities at first and died without bulbar paresis. The second case first noticed wasting of the upper limbs with bulbar symptoms, but the third had weakness of upper extremities without developing dysarthria nor dysphagia until death. These mutations account for 50% of all FALS families screened, although Cu/Zn SOD gene mutations are responsible for less than about 13-21% in the Western population. 4. Our results indicate that the progression of disease with mutations of Cu/Zn SOD is well correlated with each mutation. The exact mechanism by which the abnormal Cu/Zn SOD molecules selectively affect the function of motor neurons is still unknown. PMID- 9876870 TI - Pathophysiology of the ischemic penumbra--revision of a concept. AB - 1. The original concept of the ischemic penumbra surrounding a focus of dense cerebral ischemia is based on electrophysiological observations. In the cortex of baboons following middle cerebral artery occlusion, complete failure of the cortical evoked potential was observed at a cerebral blood flow (CBF) threshold level of approx. 0.15 ml/g/min--a level at which extracellular potassium ion activity was only mildly elevated. With a greater CBF decrement to the range of 0.06-0.10 ml/g/min, massive increases in extracellular potassium occurred and were associated with complete tissue infarction. Thus, the ischemic penumbra has been conceptualized as a region in which CBF reduction has exceeded the threshold for failure of electrical function but not that for membrane failure. 2. Recent studies demonstrate that the penumbra as defined classically by the flow thresholds does not survive prolonged periods of ischemia. The correlation of CBF autoradiograms with diffusion-weighted MR images and the regional distribution of cerebral metabolites reveals that the ischemic core region enlarges when adjacent, formerly penumbral, areas undergo irreversible deterioration during the initial hours of vascular occlusion. At the same time, the residual penumbra becomes restricted to the periphery of the ischemic territory, and its fate may depend critically upon early therapeutic intervention. 3. In the border zone of brain infarcts, marked uncoupling of local CBF and glucose utilization is consistently observed. The correlation with electrophysiological measurements shows that metabolism-flow uncoupling is associated with sustained deflections of the direct current (DC) potential resembling transient depolarizations. Such penumbral cell depolarizations, which are associated with an increased metabolic workload, induce episodes of tissue hypoxia due to the constrained collateral flow, stimulate anaerobic glycolysis leading to lactacidosis, suppress protein synthesis, and, finally, compromise energy metabolism. The frequency of their occurrence correlates with the final volume of ischemic injury. Therefore, penumbral depolarizations are regarded as a key event in the pathogenesis of ischemic brain injury. Periinfarct DC deflections can be suppressed by NMDA and non-NMDA antagonists, resulting in a significant reduction of infarct size. 4. The histopathological sequelae within the penumbra consist of various degrees of scattered neuronal injury, also termed "incomplete infarction." The reduction of neuronal density at the infarct border is a flow- and time-dependent event which is accompanied by an early response of glial cells. As early as 3 hr after vascular occlusion a generalized microglial activation can be detected throughout the ipsilateral cortex. Astrocytic activation is observed in the intact parts of the ischemic hemisphere from 6 hr postocclusion onward. Thus, the penumbra is a spatially dynamic brain region of limited viability which is characterized by complex pathophysiological changes involving neuronal function as well as well as glial activation in response to local ischemic injury. PMID- 9876872 TI - Oxygen toxicity induces apoptosis in neuronal cells. AB - 1. A high oxygen atmosphere induced apoptosis in cultured neuronal cells including PC12 cells and rat embryonic cortical, hippocampal, and basal forebrain neurons associated with DNA fragmentation and nuclear condensation. 2. The sensitivity of CNS neurons to a high-oxygen atmosphere was the following order; cortex > basal forebrain > hippocampus. 3. Cycloheximide and actinomycin-D inhibited the apoptosis, indicating that it depends on new macromolecular synthesis. In contrast, cultured postnatal CNS neurons were resistant to oxidative stress. 4. Neurotrophic factors such as nerve growth factor (NGF), fibroblast growth factor (FGF), and epidermal growth factor (EGF) blocked the apoptosis induced by a high-oxygen atmosphere. PMID- 9876875 TI - Cytokines in brain ischemia--the role of TNF alpha. AB - 1. The role of cytokines and other inflammatory mediators in the progression of ischemic brain injury is a new and exciting era of research. Evidence in support for a role for TNF alpha in this respect is emerging as evidence on de novo upregulation of TNF alpha following ischemia is now well established. 2. TNF alpha administered directly to the brain parenchyma elicits local microvascular injury in the form of pericapillary edema and leukocyte adhesion to cerebral capillaries. 3. TNF alpha administered into the cerebroventricular space prior to ischemia augment the extent of tissue damage and neurological deficits. 4. Specific and potent inhibitors of TNF alpha synthesis or TNF alpha receptors must be developed and tried to prove firmly a role for TNF alpha in ischemic brain injury. PMID- 9876874 TI - Transglutaminase-catalyzed protein cross-linking in the molecular program of apoptosis and its relationship to neuronal processes. AB - 1. One type of transglutaminase is usually accumulated in various forms of naturally occurring cell death and apoptosis. The accumulated enzyme is activated during the death process, leading to the formation of cross-linked protein structures. Degradation of the cross-linked apoptotic bodies results in the elevation of the epsilon (gamma-glutamyl)lysine isodipeptide concentration in body fluids, which may provide a diagnostic tool to monitor the apoptosis rate in various tissues under normal and pathologic conditions. 2. Extensive protein cross-linking may be directly related to the act of killing in some cells. In others, the effect of protein cross-linking is palliative, preventing leakage of macromolecules and enhancing phagocytosis of the dead cells. 3. Tissue transglutaminase has been implicated in some physiologic functions of the nervous system. 4. The molecular machinery of apoptosis is present and easily evoked in neuronal cells. 5. Effector elements of the apoptosis process have been associated with the pathogenesis of neurologic disorders. Tissue transglutaminase, representing one of the effector elements of apoptosis, may be induced and activated in cells following ischemia. It may also participate in the formation of abnormal cell inclusions and A beta deposits in amyloid plaques. PMID- 9876876 TI - Cytokine gene expression within the central nervous system. AB - 1. The identification of cytokine genes expressed in the central nervous system is critical to understanding the immune network in various diseases of brain, such as infection, degeneration, and malignancy. 2. Expression of cytokine genes in human astrocytoma cell lines and in fresh brain specimens was studied by the reverse-transcribed/polymerase chain reaction method. 3. The correlation between clinical malignancy and cytokine gene expression within malignant glioma was examined, especially regarding the relevancy of inhibitory cytokines, such as transforming growth factor-beta and interleukin-10. PMID- 9876873 TI - Free radicals as mediators of neuronal injury. AB - 1. Free radicals may play an important role in several pathological conditions of the central nervous system (CNS) where they directly injure tissue and where their formation may also be a consequence of tissue injury. 2. Free radicals produce tissue damage through multiple mechanisms, including excito-toxicity, metabolic dysfunction, and disturbance of intracellular homeostasis of calcium. 3. Oxidative stress can significantly worsen acute insults, such as ischemia, as well as chronic neurodegenerative disorders including amyotrophic lateral sclerosis (ALS) and Parkinson's disease. 4. For instance, recent findings suggest a causal role for chronic oxidative stress in familial ALS, as this disease is linked to missence mutations of the copper/zinc superoxide dismutase (SOD). 5. Thus, therapeutic approaches which limit oxidative stress may be potentially beneficial in several neurological diseases. PMID- 9876877 TI - Stress protein inductions after brain ischemia. AB - 1. Hippocampal CA1 neurons are the most vulnerable to transient cerebral ischemia. However, the mechanism has not been fully understood. 2. The mRNAs for 72-kd (HSP72) and 73-kd (HSC73) heat shock proteins (HSPs), which are located mainly in the cytoplasm, were greatly induced together in CA1 cells, with a peak at 1-2 days in gerbils. However, immunoreactive HSP72 protein was only minimally expressed in CA1 neurons. 3. The mRNA for mitochondrial HSP60 began to increase at 3 hr in CA1 cells and was sustained until 1 day. 4. The level of mRNA for cytochrome c oxidase subunit I (COX-I) progressively decreased in CA1 neurons after a transient ischemia and completely disappeared at 7 days. The activity of cytochrome c oxidase (COX) protein also showed an early decrease in CA1 cells and was followed by a reduction in the level of COX-I DNA after 2 days. 5. These results suggest that HSP gene inductions were inhibited at the translational level but that mitochondrial DNA expression was disturbed at the transcriptional level. A disturbance of mitochondrial DNA expression could cause progressive failure of energy production of CA1 cells that eventually results in neuronal cell death. PMID- 9876878 TI - Abnormalities in stress proteins in prion diseases. AB - 1. Prion diseases include kuru, Creutzfeldt-Jakob disease (CJD), Gerstmann Straussler-Scheinker disease (GSS), and fatal familia insomnia (FFI) of humans, as well as scrapie and bovine spongiform encephalopathy (BSE) of animals. 2. All these disorders involve conversion of the normal, cellular prion protein (PrPC) into the corresponding scrapie isoform (PrPSc). PrPC adopts a structure rich in alpha-helices and devoid of beta-sheet, in contrast to PrPSc, which has a high beta-sheet content and is resistant to limited digestion by proteases. That a conformational transition features in the conversion of PrPC into PrPSc implies that prion diseases are disorders of protein conformation. 3. This concept has been extended by our studies with heat shock proteins (Hsp), many of which are thought to function as molecular chaperones. We found that the induction of some Hsps but not others was profoundly altered in scrapie-infected cells and that the distribution of Hsp73 is unusual in these cells. 4. Whether the conversion of PrPC into PrPSc is assisted by molecular chaperones, or if the accumulation of the abnormally folded PrPSc is complexed with Hsps remains to be established. PMID- 9876880 TI - Indomethacin and nitrate-induced headache. PMID- 9876881 TI - The SF-36 and the assessment of HRQoL. PMID- 9876882 TI - Pathophysiology and pharmacology of migraine. Is there a place for antiemetics in future treatment strategies? AB - This article reviews the pathophysiology and pharmacology of emesis in relation to migraine pathogenesis. Also, the place of antiemetic and gastrointestinal prokinetic agents in current and future acute migraine treatment strategies is reviewed. The mechanisms of action of current and novel acute migraine therapies are considered with respect to the neurogenic and vascular hypothesis. Control of migraine-associated nausea and vomiting is often achieved with the benzamide dopamine D2 receptor antagonist metoclopramide. This drug also has 5HT3 receptor antagonist activity and reproducibly stimulates gastric motility to increase the availability of orally administered drugs. Other antiemetic and gastroprokinetic agents with potential value for the treatment of migraine-associated nausea and vomiting could speed absorption of oral antimigraine therapies without central nervous system side effects. Domperidone, a dopamine D2 receptor antagonist that does not cross the blood brain barrier is relatively free of the central side effect liability of metoclopramide. Cisapride, a benzamide 5HT4 receptor agonist gastrointestinal prokinetic drug, lacks dopamine antagonist activity. A controlled comparison of these agents as migraine co-therapies could provide information on the importance of peripheral and central mechanisms in migraine associated nausea and vomiting and improve antimigraine treatment options. PMID- 9876879 TI - Impaired motor coordination in mice lacking prion protein. AB - 1. Prion protein (PrPC) is a host-encoded glycoprotein constitutively expressed on the neuronal cell surface. Accumulation of its protease-resistant isoform is closely related to pathologic changes and prion propagation in the brain tissue of a series of prion diseases. However, the physiological role of PrPC remains to be elucidated. 2. After long-term observation, we noted impaired motor coordination and loss of cerebellar Purkinje cells in the aged mice homozygous for a disrupted PrP gene, a finding which strongly suggests that PrPC plays a role in the long-term survival of Purkinje cells. 3. We also describe the resistance of the PrP null mice to the prion, indicating the requirement of PrPC for both the development of prion diseases and the prion propagation. PMID- 9876883 TI - Migraine preventive medications: a reappraisal. AB - Newer acute care migraine medications demonstrate improved rapidity of action, consistent effectiveness, excellent safety profiles, and rarely cause rebound headaches. Their use could decrease the need for migraine-preventive medication. The present analysis derives a formula that can be used by practitioners to determine the cost-effectiveness of various migraine-preventive medications relative to selected acute-care medications. We propose a measure called the cost equivalent number (CEN), the number of headaches per month at which the cost of the preventive medication equals the cost savings in acute-care treatment realized by using the preventive medication. The use of the CEN individualizes the decision of whether to use a migraine-preventive medication, weighing both the efficacy and cost of the preventive medication against the cost of the acute care medication. A CEN lower than the migraine frequency suggests that use of a preventive medication will be cost-effective. PMID- 9876884 TI - Glycerol gangliotomy of the second dorsal cervical root in rats: an experimental study to evaluate a minimal invasive approach for the treatment of the chronic cervicogenic headache. AB - Glycerol is a known agent in the therapy of chronic tic douloureux. It has been used for about 20 years in percutaneous, retrogasserian minimal-invasive rhizotomy, although the pharmacological mechanism of the pain relief involved remains unclear. To investigate glycerol treatment as a possible replacement for invasive approaches in the therapy of chronic cervicogenic headaches, we performed an experimental study on the pathomorphologic action of anhydrous glycerol injection into the second upper cervical dorsal root ganglion (DRG) of rats. Glycerol injections into the second cervical ganglion were investigated light- and electron-microscopically in a series of 40 rats for survival times of up to 30 days. We detected an unspecific overall effect on sensory neurons and satellite cells, as well as on myelinated and unmyelinated axons and Schwann cells. This could be detected after 5 days and sometimes led to degeneration of most of the neurons. Contralateral saline injections as a control showed no morphological effects. The loss of afferent fiber connections to the posterior horn of the myelon could be detected by immunohistochemical labeling of reactive astrocytes. Our results show a glycerol-induced deterioration of the cytoarchitecture of the neurons and their glial satellite cells. The effects on the ganglion cells appear to have been mediated by membrane disturbances and loss of glial integrity. These observations are contrary to previously reported results indicating the specific effect of glycerol on thin myelinated sensory axons. PMID- 9876885 TI - Genetic abnormalities of the protein C system: shared risk factors in young adults with migraine with aura and with ischemic stroke? AB - Migraine, particularly migraine with aura (MA), may be a risk factor for ischemic stroke (IS). The reasons for this association are unknown. We investigated the presence of genetic abnormalities of the protein C system in 83 MA patients, 31 IS patients, and 124 healthy controls, all aged under 45 years. We found an increased frequency of activated protein C resistance due to Arg506Gln factor V mutation, and of protein S deficiency in both disorders, with figures higher than those reported in the general population and significantly different from those found in controls. These prothrombotic genetic abnormalities may be shared risk factors in IS and MA, and may play a role in increasing the risk of cerebrovascular disease in migraineurs. PMID- 9876886 TI - Indomethacin increases the effect of isosorbide dinitrate on cerebral hemodynamic in migraine patients: pathogenetic and therapeutic implications. AB - Intracerebral vascular reactivity induced by the nitric oxide (NO) donor isosorbide dinitrate (IDN, 5 mg sublingually) is more major and longer-lasting in migraine patients who develop delayed headache in response to the drug. The headache is purportedly due to neuronally-mediated vascular mechanisms. Indomethacin inhibits prostaglandin synthesis, which is involved in NO generation. Indomethacin also decreases cerebral blood flow by constricting precapillary resistance vessels. In the present study, the hemodynamic effects of indomethacin were evaluated in migraine patients and healthy controls by means of transcranial Doppler monitoring. Indomethacin caused a significant decrease in mean flow velocity in the middle cerebral artery. This was an additional effect to the mean velocity decrease induced by IDN. The interactions between the two drugs suggest that their effects on cerebral hemodynamics (and pain) may be of relevance both in understanding the role of NO in migraine pathogenesis and in evaluating symptomatic treatments for migraine attacks. PMID- 9876887 TI - Level of nitric oxide-dependent cGMP in patients with migraine. AB - It is believed that nitric oxide (NO) plays a significant role in migraine attacks. This molecule is formed due to the conversion of L-arginine into L citrulline. The target receptor for NO is ferrum in the heme group of cytoplasmic guanyl cyclase, the enzyme catalyzing cyclic guanosine monophosphate (cGMP) formation. To confirm this hypothesis, cGMP and nitrite level in the blood serum were measured in patients with migraine. The group under study included 37 subjects suffering from migraine with and without aura and 40 normal control subjects. The cGMP was measured during a migraine attack and 60 min following the administration of sumatriptan 6 mg subcutaneously. A statistically significant increase in cGMP level was observed in patients during a migraine attack compared to the controls. This level decreased after the administration of sumatriptan, but it was still higher than in the controls. No correlation was found between the increased cGMP level and pain intensification with clinical symptoms of migraine. The results suggest the participation of biochemical changes in migraine pathogenesis in the L-arginine-NO-cGMP pathway. PMID- 9876889 TI - Quality of life in migraine and chronic daily headache patients. AB - Primary chronic headache can affect a patient's health-related quality of life (HQL). The Medical Outcomes Study Short Form (SF-36) questionnaire has been used to address this issue. We compare the impact of headache on the HQL of patients with migraine and chronic daily headache (CDH) using the SF-36 instrument. We analyzed a group of 115 consecutive patients; 62 migraine patients and 53 CDH patients completed the questionnaire. Patterns of disability were similar between the two groups, but CDH was marked by a lower level of health scales. Patients with CDH had a significantly worse pain score in physical functioning, role functioning (physical), bodily pain, general health perceptions, and mental health than patients with migraine headache. Our results in the migraine group were similar to findings in other publications, with the lowest scores in role functioning (physical) and bodily pain. There is no previous experience in CDH patients, but the present data suggest that the SF-36 questionnaire is valuable in determining the differences in functional status among headache types. These data suggest that the SF-36 is a reliable and valid measure of the HQL of patients with CDH, and may indeed prove to be valuable in studying the efficacy of therapeutic agents for this type of headache. PMID- 9876888 TI - Significant reduction in post-lumbar puncture headache using an atraumatic needle. A double-blind, controlled clinical trial. AB - A characteristic headache occurs in about 36-55% of patients after lumbar puncture, and many of these patients need bed rest for one or more days to get relief. In a double-blind randomized trial we compared a new 22-gauge atraumatic puncture needle with the most widely used 20-gauge "conventional" needle. In 49 patients randomized to the atraumatic needle, post-lumbar puncture headache occurred in 6%, whereas in the 50 patients randomized to the conventional needle this occurred in 32% (p = 0.001). On the basis of these results we recommend use of the atraumatic needle in order to diminish the frequency of post-lumbar puncture headache. PMID- 9876890 TI - Migraine with aura shows gadolinium enhancement which is reversed following prophylactic treatment. AB - A 26-year-old patient complained of a series of migraine attacks with aura accompanied by slight pleocytosis and gadolinium (Gd-DTPA) enhancement next to the left middle cerebral artery. The migraine attacks and Gd-DTPA enhancement were reversible during prophylactic treatment with flunarizine. PMID- 9876891 TI - A review of intestinal fatty acid binding protein gene variation and the plasma lipoprotein response to dietary components. AB - OBJECTIVES: Because there is interindividual variation in the metabolic response to diet, it important to find and evaluate candidate genes, which have common functional variants in the general population, and which may be determinants of interindividual differences in the response to components of the diet, such as dietary fat and fiber. One such candidate gene is FABP2, which encodes the intestinal fatty acid binding protein. This gene has a common variation in the coding sequence, which results in either alanine or threonine to be translated at residue 54. This amino acid variation has an in vitro functional impact. In addition, there have been several published associations with metabolic phenotypes, such as impaired glucose tolerance, obesity, altered plasma lipids and lipoproteins. RESULTS: It is the FABP2 T54 variant that is almost always associated with the more deleterious phenotypic expression. Recent experiments have found that most of the inter-individual variation in the plasma lipoprotein response to dietary fiber could be attributed to the FABP2 A54T polymorphism. However, it is likely that differences in environment, such as the type of fatty acid or fiber consumed, might interact with functional differences in the gene product to produce phenotypic differences. CONCLUSIONS: This means that more experimental work, using a variety of study samples and complementary approaches, is necessary before advocating routine testing of FABP2 genotype in people in order to determine their potential responsiveness to various dietary interventions, such as increased dietary soluble fiber. PMID- 9876892 TI - Tacrolimus metabolite cross-reactivity in different tacrolimus assays. AB - OBJECTIVES: Tacrolimus (FK506) is an immunosuppressive drug with great clinical promise. There is a controversy regarding the role of tacrolimus metabolites in immunosuppression and toxicity, and immunoassays and immunophilin binding assays have not been adequately tested for metabolite cross-reactivity. Methods are limited to HPLC and HPLC-MS for quantifying the parent drug. Mixed lymphocyte culture assay (MLC) is the preferred functional bioassay for the measurement of parent drug and active metabolites but it is not practical for routine laboratory use. Due to differences in assay methods and reagent specificity, the concentration of tacrolimus in a given specimen may vary among different assay kit manufacturers. The objective of this study was to evaluate the degree of cross-reactivity or interference of the three first-generation tacrolimus metabolites [13-O-demethyl (M-I), 31-O-demethyl (M-II) and 15-O-demethyl (M-III)] among two different tacrolimus immunoassays (Immunoassay: PRO-Trac II FK506, Abbott IMx tacrolimus-II); and the radioreceptor assays (RRA) using minor immunophilins (14, 37, and 52 kDa immunophilins) and tacrolimus binding protein (FKBP12). METHODS: First-generation tacrolimus metabolites (M-I, M-II, and M-III) spiked in drug-free whole blood were assayed with RRA using three minor immunophilins (14, 37, and 52 kDa) and two commercial immunoassay procedures (Incstar PRO-Trac II tacrolimus, Abbott IMx tacrolimus II). The results were compared to previously published FKBP-12 RRA data and their immunosuppressive potency. RESULTS AND CONCLUSION: The first generation tacrolimus metabolites (M I, M-II, and M-III) were tested using concentrations of 10 and 20 ng/mL. The significance of the metabolite interference (% of the total interference) was calculated based on the relative concentration of each metabolite present at steady-state trough concentrations in renal transplant recipients (22). Metabolite I, which has no functional immunosuppressive activity showed minimal interference compared to M-II and M-III in all assays except the 14 kDa RRA. The Incstar PRO-Trac II tacrolimus assay showed the least M-I interference. Metabolite-II, which has a pharmacologic potency similar to the parent drug, showed a significant interference in the immunoassays and significant interference in radioreceptor assays. Metabolite III, which is pharmacologically inactive, produces 3-10% interference in the different assays if its presence in the blood is 6% of the parent drug. The total interference from these three metabolites was greater in the immunoassays than in the receptor assays. Receptor assays for tacrolimus provide results closer to the target value than do immunoassays. PMID- 9876893 TI - Comparison of hemoglobin A1C results by two different methods on patients with structural hemoglobin variants. AB - OBJECTIVES: The objective was to compare hemoglobin A1C (HbA1C) results obtained by two methods based on different analytical principles for individuals with a structural hemoglobin variant. DESIGN AND METHODS: Hemoglobin A1C results were obtained using the Bio-Rad Variant (based on cation exchange chromatography) and the Bayer DCA 2000 (based on an immunological reaction) on individuals with a structural hemoglobin variant. The identity of the hemoglobin variant was confirmed by high pressure liquid chromatography (HPLC) and electrophoresis. RESULTS: Hemoglobin A1C results obtained by the two methods on individuals with S, C, D, and E trait were in close agreement. CONCLUSION: The Bio-Rad Variant and Bayer DCA 2000 produce equivalent hemoglobin A1C results on patients with S, C, and E trait. With appropriate correction, correlation of hemoglobin A1C results from the Bio-Rad Variant for individuals with D trait was good (r = 0.927). Glycohemoglobin results obtained by the two methods for some unusual structural hemoglobin variants were in close agreement. PMID- 9876894 TI - A protocol for detection of mitochondrial DNA deletions: characterization of a novel deletion. AB - OBJECTIVES: To develop a protocol capable of identifying deletions in mitochondrial DNA and use it to identify the breakpoints of a mtDNA deletion in a patient with chronic progressive external ophthalmoplegia (CPEO). DESIGN AND METHODS: Deletions in mtDNA were identified by a combination of long range PCR and Southern blotting. The precise breakpoints were determined by automated DNA sequencing. RESULTS: A series of DNA samples from patients with suspected mitochondrial disease was subjected to a protocol, which combines long range PCR and Southern blotting. We found a unique deletion in a patient with CPEO and we identified the precise location of this deletion through DNA sequencing. CONCLUSIONS: Long range PCR has the advantages of speed, minimal samples requirements, and sensitivity. Southern blotting is better able to evaluate heteroplasmy and detect duplications. We suggest a protocol that enables us to identify precisely the breakpoints in a unique mutation of mtDNA in a patient with CPEO. PMID- 9876895 TI - Analytical issues on prostate-specific antigen in relation to prostate cancer screening. AB - OBJECTIVE: Evaluation of the analytical performance of the Hybritech Tandem-E PSA assay as applied to the Rotterdam part of the European Randomized Study of Screening for Prostate Cancer (ERSPC) during 1994-1997. DESIGN AND METHODS: For assessment of test performance imprecision and accuracy contributing to clinical decision making was examined. Pre-analytical variables (specimen handling), long term reagents stability and calibration check with the Stanford 90:10 PSA Calibrator were studied. RESULTS: Total prostate-specific antigen proved to be a stable marker. Provided correct storage at 4 degrees C, we were not able to find differences between samples analyzed directly after blood withdrawal and those analyzed the following day, with or without centrifugation. Day-to-day precision, we found during 1997 a coefficient of variation of 4.9% for concentration 1.8 micrograms/L and 4.2% for 4.8 micrograms/L (n = 182). The variation among the 21 Tandem-E reagent batches used during 1994-1997 proved to be small. Application of the Stanford 90:10 PSA Calibrator revealed 3% higher values for Tandem-E. CONCLUSION: The overall, long-term picture of Tandem-E PSA is reliable in our hands. Possible differences from the true PSA values, not specific for Tandem-E, need to be elaborated. PMID- 9876896 TI - Evaluation of seven PCR-based assays for the analysis of microchimerism. AB - OBJECTIVE: The presence of small numbers of cells of donor origin in the circulation of recipients of organ transplants (microchimerism) may correlate with immunologic tolerance. As part of our ongoing studies on microchimerism, we evaluated the utility of seven PCR-based assays for the detection of the less abundant DNA in paired mixtures (100 ng total DNA). DESIGN AND METHODS: DNA samples were screened to identify pairs informative for one or more PCR assays. DNA mixtures from the informative pairs were then analyzed using at least one assay. The assays were based on the X-Y homologous region; a Y chromosome microsatellite locus; three autosomal microsatellite loci; the D1S80 minisatellite locus; and sequence specific oligonucleotide probe (SSOP) analysis of the HLA DRB1 locus. RESULTS: About 0.1% of male DNA against a background of female DNA was detectable using primers for the X-Y homologous region, but the sensitivity was increased to 0.0001% using nested primers for the Y chromosome microsatellite marker. Analysis of the minor DNA component was difficult with the three autosomal microsatellite assays because of the presence of shadow bands. Similar problems with the D1S80 assay were resolved using more stringent PCR conditions, and the sensitivity was 0.1%. Using the DRB1 locus, we were able to detect 1% DNA in the mixed samples. CONCLUSIONS: These studies show that: (a) nested PCR for the Y chromosome is the most sensitive assay for the detection of microchimerism; (b) D1S80 is a useful marker for microchimerism; (c) additional optimization of analytical conditions is required if autosomal microsatellite markers and the SSOP assay are to be used for microchimerism analysis. PMID- 9876897 TI - Platelet glycoprotein IIb/IIIa genetic polymorphism is associated with plasma fibrinogen levels in myocardial infarction patients. The REGICOR Investigators. AB - Fibrinogen is the major ligand of platelet glycoprotein IIb/IIIa platelet receptor. Genes coding for platelet fibrinogen receptor glycoprotein IIb/IIIa are polymorphic. The PLA alloantigen has two antigenic determinants, PLA1 and PLA2, located in a 17-23 kD fragment of glycoprotein IIIa. We analyzed whether PLA genotype has any effect on plasma fibrinogen concentration and investigated if the effect has different magnitude in myocardial infarction patients compared with subjects free of angina or myocardial infarction. One hundred sixteen consecutive patients who suffered a myocardial infarction and 136 subjects recruited by random sampling from the local census were included in the study. PLA genotype distribution and allele frequencies in patients did not significantly differ from those in the control group. Mean fibrinogen concentration tended to be higher in controls with genotype PLA1PLA1 than in those with genotype PLA1PLA2 or PLA2PLA2, and in patients this difference reached statistical significance (p < 0.001). We conclude that the PLA polymorphism may be in linkage disequilibrium with another functional mutation in or near the promoter area of the fibrinogen gene or even in another gene, which controls the production or the clearance of fibrinogen. PMID- 9876898 TI - Endothelial cell compatibility of clarithromycin for intravenous use. AB - OBJECTIVES: Tolerance of intravenously applied clarithromycin has been tested on marginal ear veins of rabbits. Use of human umbilical venous endothelial cells (HUVEC) for testing antibiotic solutions for intravenous compatibility provides a valuable alternate model. DESIGN AND METHODS: In order to evaluate the effect of clarithromycin on intracellular purines, reflecting cell viability, energy production, signal transduction and DNA/RNA synthesis, intracellular adenosine 5' triphosphate (ATP), adenosine 5' diphosphate (ADP), guanosine 5' triphosphate (GTP), and guanosine 5' diphosphate (GDP) levels were measured by means of high performance liquid chromatography (HPLC). RESULTS: Incubation of cells with 2 mg/mL clarithromycin resulted in a rapid decrease of the intracellular ATP from 12.6 +/- 1.1 to 8.87 +/- 0.82 nmol/million cells or 1.5 +/- 0.6 nmol/million cells, after 20 or 60 min, respectively. In addition, ADP was extensively depleted. Purine nucleotide profiles were markedly different following exposure to 1 mg/mL clarithromycin. There was no significant decline of intracellular high energy phosphate levels after 20 min. CONCLUSION: These results show that clarithromycin has a better endothelial compatibility if diluted to a final concentration of 1 mg/mL. These data are in line with our clinical observations that the occurrence of phlebitis could be minimized by diluting the manufacturers' preparation of clarithromycin to 1 mg/mL. PMID- 9876899 TI - Interactions between essential and toxic elements in lead exposed children in Katowice, Poland. AB - OBJECTIVES: To determine the influence of the essential element status on blood concentrations of lead and other toxic metals. DESIGN AND METHODS: A group of 157 children from Katowice, an industrial area in Poland, was investigated for concentrations of lead and cadmium in whole blood, and mercury, selenium, zinc, copper, and magnesium in whole blood and serum. Relations between these elements, serum ferritin, hematological parameters, as well as serum selenoprotein P and glutathione peroxidase (GSH-px) were examined. Conversion factors for element concentrations (mumol to microgram): lead 207.19, cadmium 112.41, mercury 200.59, selenium 78.96, magnesium 24.31, copper 63.55, and zinc 65. RESULTS: Blood lead was negatively associated with concentrations of selenium in whole blood and serum as well as selenoprotein P and glutathione peroxidase in serum. The association was mainly apparent at low blood lead concentrations, which may indicate an influence of selenium on the kinetics of lead, rather than an effect of lead on the selenium status. Children with low serum ferritin levels had statistically higher blood cadmium levels and a tendency for higher blood lead levels, indicating increased gastrointestinal absorption of these metals at reduced iron stores. Blood lead was negatively correlated with mean corpuscular hemoglobin concentration, which may reflect the effect of lead on hemoglobin synthesis. There was an association between blood mercury and selenium, indicating a common source of intake through fish consumption. CONCLUSIONS: The results indicate that selenium and iron status may influence the kinetics of lead. PMID- 9876900 TI - Susceptibility of erythrocyte lipids to oxidation and erythrocyte antioxidant status in myocardial infarction. AB - OBJECTIVE: We evaluated the erythrocyte lipid susceptibility to oxidation and erythrocyte antioxidant status in patients with myocardial infarction (MI). DESIGN AND METHODS: Patients with MI were divided into two group according to the severity of the disease as severe (n = 30) or milder (n = 25). Malondialdehyde as a marker of lipid peroxidation was measured to show the lipid susceptibility to oxidation. Erythrocytes were stressed in vitro by hydrogen peroxide acting as the oxidative agents for 120 min. After designated time, erythrocyte MDA production was significantly higher in patients with severe MI than in controls and in patients with milder MI (p < 0.001, p < 0.001, respectively). Antioxidant status was determined by measuring the reduced glutathione levels and glutathione peroxidase activity of erythrocyte. RESULTS: In patients with MI, antioxidant status was significantly lower than in controls, and there was no significant difference between the patient groups. CONCLUSION: Determination of erythrocyte lipid susceptibility to oxidation may be a useful in vitro test to evaluate the degree of oxidative stress in myocardial infarction. PMID- 9876901 TI - Comparative study of serum and synovial fluid interleukin-11 levels in patients with various arthritides. AB - OBJECTIVE: To determine the levels of serum and synovial fluid (SF) interleukin (IL)-11 in patients with various arthritides and estimate the contribution of IL 11 to acute phase response (APR). DESIGN AND METHODS: Serum and SF IL-11 were measured by ELISA in patients with rheumatoid arthritis (RA, n = 31), seronegative spondyloarthritis (SSA, n = 23), gout (GT, n = 14) and osteoarthritis (OA, n = 20) and were correlated with ESR and acute phase proteins as well as with cytokines IL-1 alpha, IL-1 beta, IL-6, and TNF alpha. RESULTS: IL 11 was detected in both serum and SF in each group, with IL-11 being statistically higher in SF than serum in all groups, suggesting reduced catabolism or increased synthesis of IL-11 intra-articularly. Median SF IL-11 levels were higher in OA patients than in other groups and in the treated than in the untreated RA subgroup. Moreover, serum and SF IL-11 were correlated significantly with each other, and moderately with the other cytokines examined in RA, SSA, and GT, but not in OA patients, while a significant negative correlation was found with a few of the inflammatory markers examined in each group. CONCLUSIONS: Our findings provide evidence of extensive intra-articular expression of IL-11 in arthritides, especially in OA and treated RA patients, suggesting a protective role for IL-11 in joints, probably through the induction of tissue inhibitor of metalloproteinases. PMID- 9876902 TI - Parent cyclosporine in whole blood by monoclonal fluorescence polarization immunoassay for axsym and monoclonal enzyme-multiplied immunoassay for cobas fara. PMID- 9876903 TI - A candidate new gene on human chromosome 5q12 contains a motif that is found in transcriptional co-activators. PMID- 9876904 TI - Plasma corticotropin levels in patients with familial amyloidotic polyneuropathy during liver transplantation. PMID- 9876905 TI - Gadoteridol in multiple sclerosis patients. A comparison of single and triple dose with immediate vs. delayed imaging. AB - Enhancement of lesions in multiple sclerosis (MS) has been investigated using standard and high doses of gadolinium. The purposes of this study are to compare the relative merits of single and triple dose as well as examine the merits of delayed triple-dose images in a large group of patients. Thirty-seven patients with multiple sclerosis underwent contrast enhanced brain magnetic resonance imaging (MRI). After noncontrast images, a single dose (0.1 mmol/kg) of gadoteridol was administered. Subsequently, axial T1-weighted images were obtained immediately after administration, and again after a delay of approximately 20 minutes. After an additional 0.2-mmol/kg dose was administered, to provide a total cumulative dose of 0.3 mmol/kg of gadoteridol, immediate and delayed axial T1-weighted image sequences were repeated. The contrast-noise ratio (C/N) was calculated for each identified, enhancing lesion in each series. Furthermore, blinded readings were performed to determine the lesion detection rate. Of the forty definite lesions that underwent all four sequences, triple dose delayed images exhibited the highest contrast-noise ratio in a significantly larger number of lesions (p < 0.0001). Triple-dose immediate and delayed scans resulted in significantly higher contrast-noise ratios (6.47 and 9.99, respectively) when compared with those of the single dose scans (3.4 for immediate scans and 5.24 for delayed) (p < 0.01). The lesion detection rate was highest for triple dose delayed (95%), followed by triple-dose immediate (83%), single-dose delayed (68%) and finally, single-dose immediate scans (43%). Triple dose immediate was noted to have a significantly increased (p < 0.0002) lesion detection rate with respect to the standard-dose immediate scans and standard dose delayed scans (p < 0.02). In four lesions (10% of the total number of lesions), detection occurred only with the triple-dose delayed image sequence. Triple-dose 0.3 mmol/kg gadolinium with delayed imaging resulted in the highest lesion conspicuity and the highest lesion identification rate. There was a trend of progressively increasing detection rates from single-dose immediate scans to triple-dose delayed scans. Triple-dose delayed scans resulted in significantly higher (p < 2 x 10(-8) contrast noise ratios than all other sequences of this study. PMID- 9876906 TI - Congenital nasal pyriform aperture stenosis and single central maxillary incisor: CT and MRI findings. AB - The authors report five cases of congenital nasal pyriform aperture stenosis (CNPAS). Four cases are presented as neonatal nasal obstruction and also have single central maxillary incisor (SCMI). Computed tomography examination indicates three of the SCMI are unerupted and one is erupted. The fifth case has an erupted SCMI and short stature. The associated SCMI in CNPAS is believed to be more than an isolated anomaly. Awareness of the associated features of CNPAS among the radiologist and otolaryngologist may help the diagnosis and treatment of the patient. PMID- 9876907 TI - Optimization of contrast delivery for pulmonary CT angiography. AB - Pulmonary CT angiography is becoming an increasingly important tool for diagnosing Pulmonary Emboli. An important aspect in optimizing the quality of these studies is achieving maximum levels of enhancement of the pulmonary arteries. In this study we reviewed the current literature to see if there was any standardization of protocols to achieve this purpose, and found that there was no consensus. Using well described techniques for determining blood flow we were able to demonstrate the proper time delays to achieve the highest levels of enhancement. As contrast flows into the pulmonary arteries, the degree of enhancement rapidly reaches a plateau phase. This is then followed by a recirculation phase where enhancement starts to increase again. The proper time delays between scan time and start of contrast infusion should allow for enhancement to be within this recirculation phase. An experimental model was developed to demonstrate the principles. PMID- 9876908 TI - Congenital absence of the pericardium confirmed by spontaneous pneumothorax. AB - This article describes a case of complete absence of the left pericardium which was diagnosed on the chest radiograph and subsequently confirmed on computed tomography and thoracoscopy. Diagnostic confidence was bolstered by the presence of pneumopericardium which communicated via the defect with a spontaneous left pneumothorax. The clinical and imaging findings of pericardial absence are reviewed. PMID- 9876909 TI - CT features of calcifications in abdominal malignant fibrous histiocytoma. AB - The purpose of this study was to describe the computed tomographic (CT) features of intralesional calcifications in abdominal malignant fibrous histiocytomas (MFH). Forty-three pathologically proven abdominal MFH with preoperative CT were retrospectively reviewed, of which seven tumors with intralesional calcifications were studied with pathohistologic correlation. All seven calcified abdominal MFH belonged to the storiform-pleomorphic subtype with peripherally located calcifications that appeared as either lumpy (three cases) or ringlike (four cases), which were due to the presence of variable amounts of osseous (six cases) and chondroid metaplasia (two cases). About 16% of abdominal MFH, especially the storiform-pleomorphic subtype, exhibited metaplastic calcifications which were characteristically located at the periphery of the tumor and appeared as either lumpy or ringlike on CT. PMID- 9876910 TI - Aneurysms of the portal venous system. Gray-scale and color Doppler ultrasonographic findings with CT and MRI correlation. AB - Two cases of incidentally detected aneurysms involving the portal venous system are described with emphasis on gray-scale and color Doppler ultrasonographic (US) findings. Appearing on US as anechoic masses showing direct luminal continuity with the right portal vein and superior mesenteric vein, the lesions displayed spectral findings characteristic of portal venous system on color Doppler US. Dynamic helical computed tomography (CT) demonstrated simultaneous enhancement with the portal system, while the aneurysms were hypointense owing to flow void on T1-weighted spin-echo magnetic resonance (MR) images. PMID- 9876911 TI - Hepatic multi-nodular focal fatty metamorphosis in acquired porphyria cutanea tarda. Sonographic, CT, and MRI features. AB - The authors report the sonographic, computed tomography (CT), and magnetic resonance imaging (MRI) findings in a patient with acquired porphyria cutanea tarda and hepatic multi-nodular focal fatty metamorphosis. There has only been one previous description of an association between these two conditions. The diagnosis was confirmed by histological analysis of a liver biopsy specimen. Symptomatology related to the hepatic abnormality may be due to an inflammatory reaction induced by the presence of uroporphyrin crystals in the liver. Each of the radiological techniques demonstrated unusual hepatic abnormalities and, in particular, MRI showed poorly defined areas in the liver which, on T2-weighted sequences, exhibited a hypersignal with fat saturation. Treatment of porphyria cutanea tarda led to clinical remission and resolution of radiological abnormalities. PMID- 9876913 TI - Early CT findings of Fournier's gangrene in a healthy male. AB - Early clinical identification of Fournier's gangrene is imperative to avoid delay in the aggressive surgical debridement, antibiotic therapy, and sometimes hyperbaric oxygen treatments. We report on the early computed tomography findings of a non-gas-forming Fournier's gangrene in a healthy male to aid urologists, surgeons, and radiologists in the recognition of this rapidly progressive and often fatal infection. PMID- 9876912 TI - Retroaortic left renal vein with renal vein hypertension causing hematuria. AB - We describe a case of retroaortic left renal vein with hematuria. In this case, pullback pressure from the retroaortic left renal vein to the inferior vena cava revealed left renal vein hypertension according to criteria of the "nutcracker phenomenon." We stress that left renal vein anomaly including retroaortic left renal vein can cause clinical symptoms such as hematuria. PMID- 9876914 TI - Giant-cell tumor of the tendon sheath arising from the posterior cruciate ligament of the knee: a case report and review of the literature. AB - The localized form of a giant-cell tumor of the tendon sheath is uncommon and rarely intraarticular. While the sonographic findings are nonspecific, the magnetic resonance (MR) findings can be diagnostic. Relatively homogeneous iso- or marginal hyperintensity on T2-weighted sequences, lack of susceptibility effects due to hemosiderin, the demonstration of a fibrous capsule and the absence of adjacent joint involvement may help to differentiate localized and diffuse forms of giant-cell tumors of tendon sheaths and pigmented villonodular synovitis. PMID- 9876915 TI - NMR analysis of cbEGF domains gives new insights into the structural consequences of a P1148A substitution in fibrillin-1. AB - Fibrillin-1 is a modular glycoprotein and a major component of the 10-12 nm microfibrils of the extracellular matrix. Mutations in the fibrillin-1 (FBN 1) gene result in the connective tissue disease the Marfan syndrome (MFS) and related disorders. The calcium binding EGF-like (cbEGF) domain is the predominant structural motif of the protein and >70% of mutations leading to MFS disrupt this domain. A missense mutation which changes a proline to alanine (P1148A) in cbEGF domain 13 has been associated with a number of fibrillin disorders including MFS and Shprintzen-Goldberg syndrome. However, it has also been described as a polymorphism. In this study comparative NMR analyses on wild-type and mutant forms of covalently-linked fibrillin cbEGF domain pairs have been performed to investigate the structural consequences of this substitution. A comparison of the two-dimensional NOESY spectra of the wild-type and mutant forms of cbEGF domains 12 & 13 and cbEGF domains 13 & 14 indicated that the proline to alanine amino acid change does not introduce a significant structural defect into cbEGF domain 13 or the adjacent domains and most likely represents a polymorphism. These results demonstrate how, in the case of a protein with a well defined domain organisation such as fibrillin-1, comparative NMR analyses can be used to substantiate genetic evidence for the polymorphic status of an amino acid. PMID- 9876916 TI - Prediction of membrane proteins based on classification of transmembrane segments. AB - The number of transmembrane segments often corresponds to a structural or functional class of membrane proteins such as to seven-transmembrane receptors and six-transmembrane ion channels. We have developed a new prediction method to detect the membrane protein class that is defined by the number of transmembrane segments, as well as to locate the transmembrane segments in the amino acid sequence. Each membrane protein class is represented by a model of ordering different types of transmembrane segments. Specifically, we have classified the transmembrane segments in known membrane proteins into five groups (types) using the Mahalanobis distance with the average hydrophobicity and the periodicity of hydrophobicity as a measure of similarity. The discriminant functions derived for these groups were then used to detect transmembrane segments and to match with the models for one- to fourteen-spanning membrane proteins and for globular proteins. Using the test data set of 89 membrane proteins whose transmembrane positions are known by experimental evidence, 61.8% of the proteins and 85.1% of the transmembrane segments were correctly predicted. Because of the new feature to predict membrane protein classes, the method should be useful in the functional assignment of genomic sequences. PMID- 9876917 TI - Prediction of the helix/strand content of globular proteins based on their primary sequences. AB - An improved multiple linear regression method has been proposed to predict the content of alpha-helix and beta-strand of a globular protein based on its primary sequence. The amino acid composition and the auto-correlation functions based on the hydrophobicity profile of the primary sequence have been taken into account in the algorithm. The resubstitution test shows that the average absolute errors are 0.077 and 0.073 with the standard deviations 0.059 and 0.057 for the prediction of the content of alpha-helix and beta-strand, respectively. A stringent cross-validation test, i.e., the jackknife test, shows that the average absolute errors are 0.087 and 0.081 with the standard deviations 0.067 and 0.065 for the prediction of the content of alpha-helix and beta-strand, respectively. Both tests indicate the self-consistency and the extrapolating effectiveness of the new algorithm. This greatly improves on previous results (Eisenhaber,F., Imperiale,F., Argos,P. and Frommel,C., 1996, Proteins, 25, 157-168). Compared with other methods currently available, our method has the merits of simplicity and ease-of-use as well as a higher prediction accuracy. The only input of the method is the primary sequence of the query protein to be predicted. The program is available on request via e-mail: ctzhang@tju.edu.cn. PMID- 9876918 TI - Novel method to detect a motif of local structures in different protein conformations. AB - In order to detect a motif of local structures in different protein conformations, the Delaunay tessellation is applied to protein structures represented by C(alpha) atoms only. By the Delaunay tessellation the interior space of the protein is uniquely divided up into Delaunay tetrahedra whose vertices are the C(alpha) atom positions. Some edges of the tetrahedra are virtual bonds connecting adjacent residues' C(alpha) atoms along the polypeptide chain and others indicate interactions between residues nearest neighbouring in space. The rules are proposed to assign a code, i.e., a string of digits, to each tetrahedron to characterize the local structure constructed by the vertex residues of one relevant tetrahedron and four surrounding it. Many sets comprised of the local structures with the same code are obtained from 293 proteins, each of which has relatively low sequence similarity with the others. The local structures in each set are similar enough to each other to represent a motif. Some of them are parts of secondary or supersecondary structures, and others are irregular, but definite structures. The method proposed here can find motifs of local structures in the Protein Data Bank much more easily and rapidly than other conventional methods, because they are represented by codes. The motifs detected in this method can provide more detailed information about specific interactions between residues in the local structures, because the edges of the Delaunay tetrahedra are regarded to express interactions between residues nearest neighbouring in space. PMID- 9876919 TI - Determinants of side chain conformational preferences in protein structures. AB - A discriminatory function based on a statistical analysis of atomic contacts in protein structures is used for selecting side chain rotamers given a peptide main chain. The function allows us to rank different possible side chain conformations on the basis of contacts between side chain atoms and atoms in the environment. We compare the differences in constructing side chain conformations using contacts with only the local main chain, using the entire main chain, and by building pairs of side chains simultaneously with local main chain information. Using only the local main chain allows us to construct side chains with approximately 75% of the chi1 angles within 30 degrees of the experimental value, and an average side chain atom r.m.s.d. of 1.72 A in a set of 10 proteins. The results of constructing side chains for the 10 proteins are compared with the results of other side chain building methods previously published. The comparison shows similar accuracies. An advantage of the present method is that it can be used to select a small number of likely side chain conformations for each residue, thus permitting limited combinatorial searches for building multiple protein side chains simultaneously. PMID- 9876920 TI - A cooperative folding unit in HIV-1 protease. Implications for protein stability and occurrence of drug-induced mutations. AB - We investigated the HIV-1 protease molecule for the occurrence of cooperative folding units, i.e. structural units that exhibit a relatively stronger protection against unfolding than do other parts of the molecule. Calculated unfolding penalties are used to delineate folding units. This procedure identifies a folding core in HIV-1 protease, based on an ensemble of denatured states derived from native structures, comprising a spatially close unit of residues 84-91, 74-78 and 22-32, the last of which contains the active site residues D25, T26 and G27. Observed enzyme mutations of HIV-1 protease, either naturally occurring or induced by drug therapy, are found in regions that are not structurally designed to withstand unfolding. These mutations are especially likely to occur in the flap region, a part of the protein which is not essential for the stability of the protein, but does contribute significantly to the stability of protease-drug complexes. A similar avoidance of structurally protected regions in the reverse transcriptase enzyme is also observed. PMID- 9876921 TI - Three-dimensional structures of the cysteine proteases cathepsins K and S deduced by knowledge-based modelling and active site characteristics. AB - Human cathepsins K and S are recently identified proteins with high primary sequence homology to members of papain superfamily, including cathepsins B, L, H and papain. Models of the tertiary structures of cathepsins K and S and their complexes with a specific substrate and inhibitor were constructed and compared with the recently determined X-ray structure of cathepsin K. A major problem in the determination of the three-dimensional structure of proteins concerns the quality of the structural models obtained from the interpretation of experimental data. The framework of the tertiary structures of cathepsins K and S consisted of structurally conserved regions from the tertiary structure of the papain superfamily and the variable regions were constructed with fragments of other proteins from the protein data base. Based on docking studies the non-bonded interaction energies of ligands with the cathepsins were estimated. These energies correlate with experimentally determined substrate and inhibitory potency. PMID- 9876922 TI - Geometric invariant core for the V(L) and V(H) domains of immunoglobulin molecules. AB - A new algorithmic method for identifying a geometric invariant of protein structures, termed geometrical core, is developed. The method used the matrix of C(alpha)-C(alpha) distances and does not require the usual superposition of structures. The result of applying the algorithm to 53 immunoglobulin structures led to the identification of two geometrical core sets of C(alpha) atoms positions for the V(L) and V(H) domains. Based on these geometric invariants a preferred coordinate system for the immunoglobulin family is constructed which serves as a basis for structural prediction. The X-ray atom coordinates for all available immunoglobulin structures are transformed to the preferred coordinate system. An affine symmetry between the V(L) and V(H) domains is defined and computed for each of the 53 immunoglobulin structures. PMID- 9876923 TI - Molecular dynamics simulations of HLA-DR4 (DRB1*0405) complexed with analogue peptide: conformational changes in the putative T-cell receptor binding regions. AB - The specific recognition of foreign peptide bound to the major histocompatibility complex (MHC) molecule by T-cell receptor (TCR) leads to T-cell activation. We found that analogue peptides containing single amino acid substitutions at the third amino acid position (p3), p5, p7 and p8 of the index peptide (YWALEAAAD) induced different response patterns of T cell clones specific for the index peptide in the context of the human MHC class II molecule HLA-DR4. Analogue peptides were classified into three types, agonists, antagonists or null peptides (non-agonistic and non-antagonistic peptides). A molecular basis for how these slight changes lead to such different consequences for T cells has not been described. To explore the mechanistic basis of these observations, molecular dynamics simulations at 300 K of 300 ps duration were carried out for the DR4 index peptide, DR4-agonist, and DR4-antagonist complexes. The simulations showed that the DR4-antagonist complexes were distinguished from the DR4-index peptide and DR4-agonist complexes by relatively higher deviations of C(alpha) atoms in proposed TCR-binding regions, suggesting that subtle changes of the exposed framework of the peptide binding groove by the antagonist peptides could induce the TCR antagonistic activities. PMID- 9876924 TI - Molecular modelling investigation of wild-type and the R528H mutated segment IIS4 of human L-type voltage-gated calcium channels. AB - A molecular modelling study was performed in order to investigate the pathologically modified properties of L-type voltage-gated calcium channels caused by the arginine-to-histidine mutation at position 528 (R528H) in segment IIS4. For an appropriate consideration of the ionization state the finite difference Poisson-Boltzmann method was applied to compute the apparent pKa values of all titratable residues using standard conditions and an explicit lipophilic environment, respectively. Restrained molecular dynamics simulations were carried out for the alpha-helical transmembrane segments of the wild-type and the R528H mutant to explore their conformational behaviour. While both structures showed almost the same side-chain flexibility around the conserved residues, only the mutant partially formed a hydrogen bond from H528 to R531 during dynamics simulations. This local interaction not only causes a lower mobility of the directly involved residues but also leads to a global distortion of all positively charged amino acids of the mutant. Mostly affected is the side chain of R534 that is shifted about 61 degrees closer to R531 and about 5 A in the direction of the cytoplasm. Subsequent examination of the molecular characteristics of this putative voltage sensor of the channel revealed considerable variations with regard to hydrogen bonding and electronic properties. Most obvious are the dramatic loss of a strong positive molecular electrostatic potential and the reduced hydrogen donor activities around position 528 of the mutant. How these results may be interpreted in relation to an enhanced inactivation rate is discussed, considering earlier findings at homologous voltage-gated potassium and sodium channels. PMID- 9876926 TI - An isoleucine zipper peptide forms a native-like triple stranded coiled coil in solution. AB - Recent studies in the field of de novo protein design have focused on the construction of native-like structures. Here we describe the design and characterization of an isoleucine zipper peptide intended to form a parallel triple-stranded coiled coil. To obtain the native-like structural uniqueness, the hydrophobic interface of the peptide consists of beta-branched Ile residues for complementary side chain packing. The peptide forms a stable triple-stranded coiled coil, as determined by circular dichroism and sedimentation equilibrium analyses. A fluorescence quenching assay after the incorporation of acridine revealed a parallel orientation of the peptides. The structural uniqueness of the coiled coil was confirmed by proton-deuterium amide hydrogen exchange and hydrophobic dye binding. The peptide contains amide protons with hydrogen exchange rates that are approximately an order of magnitude slower than those expected if the exchange occurred via global unfolding. A hydrophobic dye does not bind to the peptide. These results strongly suggest that the peptide folds into a well-packed structure that is very similar to the native state of a natural protein. Thus, Ile residues in the hydrophobic interface can improve the side chain packing, which can impart native-like structural uniqueness to the designed coiled coil. PMID- 9876925 TI - Structure-function analysis of a conserved aromatic cluster in the N-terminal domain of human epidermal growth factor. AB - The importance of a cluster of conserved aromatic residues of human epidermal growth factor (hEGF) to the receptor binding epitope is suggested by the interaction of His10 and Tyr13 of the A-loop with Tyr22 and Tyr29 of the N terminal beta-sheet to form a hydrophobic surface on the hEGF protein. Indeed, Tyr13 has previously been shown to contribute a hydrophobic determinant to receptor binding. The roles of His10, Tyr22 and Tyr29 were investigated by structure-function analysis of hEGF mutant analogues containing individual replacements of each residue. Substitutions with aromatic residues or a leucine at position 10 retained receptor affinities and agonist activities similar to wild-type indicating that an aromatic residue is not essential. Variants with polar, charged or aliphatic substitutions altered in size and/or hydrophobicity exhibited reduced binding and agonist activities. 1-Dimensional 1H NMR spectra of high, moderate and low-affinity analogues at position 10 suggested only minor alterations in hEGF native structure. In contrast, a variety of replacements were tolerated at position 22 or 29 indicating that neither aromaticity nor hydrophobicity of Tyr22 and Tyr29 is required for receptor binding. CD spectra of mutant analogues at position 22 or 29 indicated a correlation between loss of receptor affinity and alterations in hEGF structure. The results indicate that similar to Tyr13, His10 of hEGF contributes hydrophobicity to the receptor binding epitope, whereas Tyr22 and Tyr29 do not appear to be directly involved in receptor interactions. The latter conclusion, together with previous studies, suggests that hydrophobic residues on only one face of the N-terminal beta-sheet of hEGF are important in receptor recognition. PMID- 9876928 TI - Role of free Cys121 in stabilization of bovine beta-lactoglobulin B. AB - Mixed disulfide derivatives of bovine beta-lactoglobulin (BLG) were studied by circular dichroism (CD), gel-permeation HPLC and high-sensitivity differential scanning calorimetry (HS-DSC). It was shown that modification of Cys121 with mercaptopropionic acid and mercaptoethanol does not affect the secondary structure of BLG, but results instead in tertiary and quaternary structure changes. At neutral pH, the equilibrium dimer<==>monomer of modified beta lactoglobulin is shifted towards monomeric form. In contrast to native BLG, thermal denaturation of modified beta-lactoglobulin is fully reversible in neutral and acidic pH as demonstrated by CD and HS-DSC measurements. Modification of Cys121 results in a significant decrease of transition temperature (-6 degrees C) and enthalpy (-106 kJ/mol) at pH 2.05 while unfolding heat capacity increment remains unchanged. Thermal unfolding transitions of native and modified beta lactoglobulin at pH 2.05 are well approximated by a two-state model suggesting that no intermediate states appear after modification. The difference in Gibbs energy of denaturation between native and modified beta-lactoglobulin, 8.5 kJ/mol at 37 degrees C and pH 2.05, does not depend on the nature of the introduced group (charged or neutral). Computer analysis of possible interactions involving Cys121 in a three-dimensional structure of beta-lactoglobulin revealed that the thiol group is too far away from neighboring residues to form side-chain hydrogen bonds. This suggests that the sulfhydryl group of Cys121 may contribute to the maintenance of BLG tertiary structure via water mediated H-bonding. PMID- 9876927 TI - Effect of replacing a conserved proline residue on the function and stability of bovine adrenodoxin. AB - A proline residue in the C-terminal part of the polypeptide chain is highly conserved among many [2Fe-2S] ferredoxins. To investigate the requirement for proline at this position, we constructed steric (4-108W), charged (4-108K), polar (4-108S) and non-polar (4-108A) truncated mutants of adrenodoxin and studied them for biological function and stability. Although the variants were expressed in Escherichia coli with a significantly lower yield compared with wild-type adrenodoxin, successful incorporation of the iron-sulfur cluster suggested their proper folding. Similar absorption, CD and EPR spectra indicated that the cluster environment was not affected by the mutations. No evidence for an essential role of Pro108 in determining the redox potential of adrenodoxin or its interactions with the redox partners was found. However, replacement of this residue results in a dramatic decrease in the overall protein stability. The differences in the Gibbs energy of unfolding at 37 degrees C, delta[delta(d)G(37 degrees C)], are 5.0, -7.8, -10.1 and -10.7 kJ/mol for 4-108A, 4-108S, 4-108W and 4-108K mutants, respectively, compared with 4-108P as a control. We conclude that the principle function of Pro108 is to stabilize adrenodoxin threefold: (i) through limitation of the conformation of the polypeptide chain in this region, (ii) through a hydrogen bond to Arg14 and (iii) favorable hydrophobic contacts. PMID- 9876929 TI - Separation of the two reactions, oxidation and isomerization, catalyzed by Streptomyces cholesterol oxidase. AB - Site-directed mutagenesis was used to identify key amino acid residues of the cholesterol oxidase from Streptomyces sp., which catalyzes the oxidation of cholesterol and the isomerization of 5-cholesten-3-one. Eight mutant enzymes were constructed and the following amino acid substitutions were identified: N318A, N318H, E356A, E356D, H441A, H441N, N480A and N480Q. Mutants N318A and N318H retained both oxidation and isomerization activities. The mutant E356D retained oxidation but not isomerization activity. On the other hand, mutants N480A and N480Q showed no oxidation activity but retained their isomerization activities. The two catalytic reactions, oxidation and isomerization, in cholesterol oxidase were thus successfully separated. When the H441A or H441N mutation was introduced, both the oxidase and isomerase activities were completely lost. The H441, E356 and N480 residues thus appear to participate in the catalysis of cholesterol oxidase, whereas N318 does not. An analysis of the products of these mutant enzymes suggested that the previously proposed 6-hydroxylation reaction by cholesterol oxidase is actually autooxidation from 5-cholesten-3-one. Kinetic studies of the purified wild-type and mutant enzymes showed that the k(cat)/Km values for oxidation in E356D and for isomerization in N480A increased six- and threefold, respectively, over those in the wild-type. These mutational effects and the reaction mechanisms are discussed in terms of the three-dimensional structure of the enzyme constructed on the basis of homology modeling. PMID- 9876930 TI - Use of fortuitous in vitro mutations in a synthetic Cu-Zn superoxide dismutase gene to illuminate protein structure-function relationships. AB - A completely synthetic bovine copper-zinc superoxide dismutase gene (Cu-ZnSOD), designed using the most favoured codons for expression in yeast, was constructed. Fortuitous mutations introduced while cloning the synthetic gene permitted the additional construction of four altered-polypeptide products representing two single (Pro121-->Leu and Gly128-->Asp), one double (Pro100-->Leu, Arg113-->Lys) and one triple (Pro100-->Leu, Arg113-->Lys, Pro121-->Leu) mutant. All five versions of the gene were expressed in a SOD-deficient Escherichia coli strain. The 'wild-type' version of the gene and the two single-mutants were expressed to equal extents (approximately 8% of total soluble protein). However, compared with the 'wild-type' enzyme, one single-mutant (Gly128-->Asp) showed almost twice as much dismutase activity whilst the other (Pro121-->Leu) exhibited only 70% of the 'wild-type' level. In contrast, the double and triple mutants showed diminished expression of the gene (approximately 1 and 3% of total soluble protein, respectively) and almost no detectable SOD activity. Polyclonal antibovine SOD antibody bound all the recombinant proteins, although some of the products showed decreased size and probably altered conformations. The 'wild-type' superoxide dismutase recombinant was correctly dimerized and possessed dismutase activity, as did the Gly128-->Asp mutant despite the change in charge. Mutations in the other three versions affected enzyme folding and activity. The effect of the different mutations appeared to be additive, with the Pro121-->Leu substitution leading to the apparent proteolytic degradation of the enzyme in vivo. PMID- 9876931 TI - The role of surface lysines in pyrimidine (6-4) pyrimidone photoproduct binding by a high-affinity antibody. AB - The roles of four VH surface lysine side-chains in antibody recognition of DNA containing a central pyrimidine (6-4) pyrimidone photoproduct was investigated. Three of the four lysines (at positions H62, H64 and H66) are strongly conserved; however, the presence of lysine at position H68 was highly unusual. Each lysine was replaced individually by alanine; in addition, a quadruple mutant was also created. Studies of the binding kinetics revealed that these lysine residues mainly influenced the association phase. Our results suggest that these side chains help to guide the DNA polyanion to the antigen binding pocket by electrostatic effects. PMID- 9876932 TI - The interactions of the cytokine-binding homology region and immunoglobulin-like domains of gp130 with oncostatin M: implications for receptor complex formation. AB - The receptor gp130 is utilized by cytokines including interleukin 6, leukemia inhibitory factor, oncostatin M, cilary neurotrophic factor and cardiotrophin. It is essential for myocardial development and haematopoiesis during embryogenesis, and its role as a shared signal transducer among different cytokines explains their overlapping biological functions. Although gp130 contains a cytokine binding homology region (CHR) analogous to the extracellular growth hormone receptor, the complexes that utilize gp130 are not simple dimerizations of receptors around a single cytokine but involve receptor interactions with additional sites on the ligand resulting in higher order complexes. Analysis by surface plasmon resonance of the binding of the immunoglobulin-like and CHR domains of the extracellular portion of gp130 to mutants of the cytokine oncostatin M reveal that the CHR forms the main binding site for oncostatin M by a classical site II interaction, but in addition a second interaction occurs involving the receptor's immunoglobulin-like domain and the cytokine's site III at the N-terminus of the D helix. The implications for complex formation are discussed. PMID- 9876933 TI - Stepwise transplantation of an active site loop between heat-labile enterotoxins LT-II and LT-I and characterization of the obtained hybrid toxins. AB - Members of the cholera toxin family, including Escherichia coli heat-labile enterotoxins LT-I and LT-II, catalyze the covalent modification of intracellular proteins by transfer of ADP-ribose from NAD to a specific arginine of the target protein. The ADP-ribosylating activity of these toxins is located in the A subunit, for which LT-I and LT-II share a 63% sequence identity. The flexible loop in LT-I, ranging from residue 47 to 56, closes over the active site cleft. Previous studies have shown that point mutations in this loop have dramatic effects on the activity of LT-I. Yet, in LT-II the sequence of the equivalent loop differs at four positions from LT-I. Therefore five mutants of the active site loop were created by a stepwise replacement of the loop sequence in LT-I with virtually all the corresponding residues in LT-II. Since we discovered that LT-II had no activity versus the artificial substrate diethylamino-benzylidine aminoguanidine (DEABAG) while LT-I does, our active site mutants most likely probe the NAD binding, not the arginine binding region of the active site. The five hybrid toxins obtained (Q49A, F52N, V53T, Q49V/F52N and Q49V/F52N/V53T) show (i) great differences in holotoxin assembly efficiency; (ii) decreased cytotoxicity in Chinese hamster ovary cells; and (iii) increased in vitro enzymatic activity compared with wild type LT-I. Specifically, the three mutants containing the F52N substitution display a greater Vmax for NAD than wild type LT I. The enzymatic activity of the V53T mutant is significantly higher than that of wild type LT-I. Apparently this subtle variation at position 53 is beneficial, in contrast to several other substitutions at position 53 which previously had been shown to be deleterious for activity. The most striking result of this study is that the active site loop of LT-I, despite great sensitivity for point mutations, can essentially be replaced by the active site loop of LT-II, yielding an active 'hybrid enzyme' as well as 'hybrid toxin'. PMID- 9876935 TI - Promise and peril: new genetic discoveries and Alzheimer disease. PMID- 9876934 TI - The diphtheria toxin transmembrane domain as a pH sensitive membrane anchor for human interleukin-2 and murine interleukin-3. AB - We have constructed two fusion proteins T-hIL-2 and T-mIL-3 in which human interleukin-2 (hIL-2) or murine interleukin-3 (mIL-3) are fused to the C-terminus of the diphtheria toxin transmembrane domain (T domain). Two additional fusion proteins, T-(Gly4-Ser)2-hIL-2 and T-(Gly4-Ser)2-mIL-3, were derived by introduction of the (Gly4-Ser)2 spacer between the T domain and cytokine components. Recognition of the hIL-2 receptor or the mIL-3 receptor by the corresponding recombinant proteins was demonstrated by their capacity to stimulate cytokine-dependent cell lines. All proteins retained the capacity of the T domain to insert into phospholipid membranes at acidic pH. Finally, anchoring of both cytokines to the membrane of lipid vesicles or living cells was assessed by specific antibody recognition. Our results show that the T domain fused to the N-terminus of a given protein can function as a pH sensitive membrane anchor for that protein. PMID- 9876936 TI - Genetic testing and Alzheimer disease: the promise. PMID- 9876937 TI - Gene-environment interaction in late-onset Alzheimer disease: the role of apolipoprotein-epsilon4. PMID- 9876938 TI - Genetic testing and Alzheimer disease: ethical issues for providers and families. PMID- 9876939 TI - Alzheimer dementia: the Judaeo-Biblical perspective on patient care and genetic predestination or neurocalvinism. PMID- 9876940 TI - Alzheimer disease: where are we now? Where are we going? PMID- 9876941 TI - Genetic testing for Alzheimer disease: a view from Washington. PMID- 9876942 TI - Utilization of healthcare resources for asthma in Singapore: demographic features and trends. AB - This study examined the healthcare utilization patterns for asthma in Singapore, a tropical island city-state, between 1986 and 1993. Asthma was ranked fifth among principal conditions with the highest number of discharges in Singapore (2.4 discharges per 1,000 population per annum). Among the 0-14 year-olds, asthma was ranked second, only after accidents and injuries, as the condition with the highest number of discharges (5.2 per 1,000). It was estimated that there were approximately 20,000 accident and emergency (A & E) room visits for asthma per annum (7.0 visits per 1,000 population per annum). There were, however, no significant changes in the number of hospitalized or emergency room cases for asthma over the period of analysis. In the primary healthcare setting, asthma ranked 6th among the leading conditions seen by the general practitioners and government polyclinics (15 visits per 100 population per annum). Among under 15 year olds, asthma (37 visits per 100 age-adjusted population per annum) ranked as the second leading condition behind upper respiratory tract infections. This study has shown that asthma causes a significant amount of morbidity in the Singapore community. PMID- 9876943 TI - The value of a single skin prick testing for specific IgE Dermatophagoides pteronyssinus to distinguish atopy from non-atopic asthmatic children in the tropics. AB - In a tropical setting, where the prevalence of house dust mites (Dermatophagoides pteronyssinus) is high, we examined the advantage of a single battery of skin prick testing (SPT) for mite as a diagnostic tool by comparing the results of radio-allergo-sorbent-test (RAST) to distinguish allergic from non-allergic asthma in children. Fifty asthmatic children were enrolled in this study. After questioning the parents, SPT were carried out using house dust mite (D. pteronyssinus) and other 9 common aero-allergens and blood were taken for measuring the total IgE (PRIST) and specific IgE for mite (RAST). Dust was obtained from 14 asthmatic children's houses and mite counting was done under a high power microscope. With a daily temperature of 27.0 +/- 0.5 degrees C and a relative humidity of 80 +/- 1%, house dust mites were found in all samples; and 81% of the allergic asthmatic children had positive SPT for D. pteronyssinus. SPT for D. pteronyssinus had a sensitivity of 95% and specificity of 52% using RAST as gold standard and there was a moderate positive correlation between the size of SPT wheals and RAST scores for D. pteronyssinus (r = 0.67 and p = 0.001). The findings of this study suggest that SPT for mites should be used as a screening test and positive SPT should be confirmed by RAST. PMID- 9876944 TI - Decreased CD4+/CD8+ ratio in major type of recurrent aphthous ulcers: comparing major to minor types of ulcers. AB - The etiology of recurrent aphthous ulcers (RAU) has not been clearly defined. However, the results of several studies indicated the evidence of the role of immunological factors. The association between the regulator and effector component of the immune system in RAU needs clarifying by comparing major and minor type of RAU patients. The proportion of peripheral blood lymphocyte subsets were enumerated during active ulcer phase and analyzed in relation to ulcer types. Nineteen patients with RAU (12 minor type and 7 major type) and 8 healthy volunteers, of both sexes, aged 24-54 years old were tested. CD3+ (T cell), CD4+ (helper T cell), CD8+ (suppressor/cytotoxic T cell), CD19+ (B cell), and CD16+/CD56+ (NK cell) were determined by using appropriate monoclonal antibodies in double colored flow cytometry. The results showed that CD4+ was lower in RAU than control (P < 0.01). Comparing both types of RAU, it appeared that CD8+ was higher in the major type than the minor type (p < 0.01); CD4+/CD8+ ratio in the major type was lower than the minor type (P < 0.01). There was no difference in CD19+ and CD16+/CD56+ between any groups compared. The finding indicated that RAU was associated with abnormal proportions of CD4+ and CD8+ cells which was dependent on the severity of the lesion. PMID- 9876945 TI - Decreased nailfold capillary density in limited scleroderma with pulmonary hypertension. AB - Approximately 20% of patients with the limited form of scleroderma will develop pulmonary hypertension which is generally a late stage fatal complication. Why pulmonary hypertension occurs in this subset of patients is unknown and it has not been possible to predict which patients are at risk. Nailfold capillary dilatation, distortion and drop occurs universally in patients with scleroderma and is generally an early finding. The present study was conducted to investigate whether quantitative nailfold capillaroscopy could distinguish those limited scleroderma patients who have established pulmonary hypertension. Quantitative nailfold capillaroscopy was performed by Visual Image Analysis in 10 healthy subjects and 20 patients with limited scleroderma (18 centromere +ve), of whom 8 had established pulmonary hypertension. It was found that scleroderma patients with pulmonary hypertension had a significant reduction in capillary density compared with patients lacking this complication (p < 0.01). Patients with scleroderma have significantly more dilated capillaries than controls although no significant differences were observe between the two patient subgroups. The finding of reduced nailfold capillary density in scleroderma patients with established pulmonary hypertension has possible pathogenic significance and may allow detection of this subgroup at an early stage in their disease progression. PMID- 9876946 TI - Liposome entrapped allergen reduces plasma histamine in sensitized mice. AB - Immunotherapy of allergic diseases is associated with problems of adverse systemic reactions. We have shown earlier that liposome entrapped allergen (LEA) is effective in inducing IgG response and restricting IgE response in immunized mice. This mode of treatment may be more effective and safer if it can prevent anaphylaxis. To determine this feature, mice were administered allergen preparations repeatedly and later challenged with the same allergen. Mice given liposomal preparation showed lower specific IgE response as compared to the mice given free allergen or alum adsorbed allergen of Artemisia scoparia. Specific IgG response was higher in mice immunized with LEA. The mice immunized with liposomal preparation survived whereas others injected with free allergen or alum adsorbed allergen died probably due to anaphylaxis. High levels of histamine were observed in mice injected with free allergen as compared to the mice injected LEA. The increase in plasma histamine level may be the cause of anaphylaxis during allergen challenge. In conclusion, LEA could be used as a safe and effective mode of immunotherapy for allergy diseases, since it reduces plasma histamine levels considerably thereby reducing the chances of anaphylaxis. PMID- 9876947 TI - Hepatocellular carcinoma: significance of HBV vertical transmission. AB - In two cases of childhood hepatocellular carcinoma in Thailand, we established vertical transmission of hepatitis B virus infection as the underlying cause. With the first patient, the family history of HBV carriage became evident and a pedigree could be devised which demonstrated the high prevalence among the family members and hence evidence of vertical transmission. In the case of the second patient, we performed PCR and subsequent direct sequencing of HBV DNA isolated from his HBsAg-positive mother's, as well as from his serum, comparing the nucleotide sequences with those of a pregnant woman diagnosed as an asymptomatic HBV carrier, of another asymptomatic HBV carrier and of a reference strain, respectively, all belonging to the same genotype and subtype as the samples tested. Our results clearly indicate the necessity for nation-wide hepatitis B vaccination starting at birth, at least in hyperendemic areas like the Far East, in order to forestall HBV carriage and ensuing cirrhosis and/or HCC by preventing vertical transmission. PMID- 9876948 TI - Circulating interleukin (IL)-1 beta, IL-6 and tumor necrosis factor-alpha in children with febrile infection--a comparison with C-reactive protein. AB - Circulating interleukin (IL)-1 beta, IL-6, and tumor necrosis factor (TNF)-alpha were examined in 42 febrile children with fever lasting more than 4 days. Their diagnosis were probable viral syndrome in 22, urinary tract infection (UTI) in 10, and probable bacterial pneumonia in 10. None of our study patients had detectable serum IL-1 beta. TNF-alpha levels were significantly higher in children with pneumonia than in those with viral syndrome (p < 0.01). Children with UTI and pneumonia had significantly higher IL-6 and CRP, compared to those with probable viral syndrome (p < 0.01 for both IL-6 and CRP). When appropriate cutoff values are chosen, IL-6 had greatly improved specificity (86.4%, > 20 pg/ml) to demonstrate UTI and pneumonia, as compared to that using CRP (48%, > 40 mg/l). After three days' antibiotic treatment, IL-6 fell to control levels in children with UTI and pneumonia, while CRP remained elevated. There was no difference in TNF-alpha values before and after treatment. Thus, IL-6, rather than IL-1 beta and TNF-alpha, may be a helpful diagnostic tool for evaluation of pediatric febrile infection. Sequential studies involving more patients are needed to determine whether IL-6 is better than CRP in this clinical setting. PMID- 9876949 TI - Comparison of the reactogenicity and immunogenicity of two different dose levels of hepatitis A vaccine in healthy children and adolescents. AB - An open study was performed to compare the reactogenicity and immunogenicity of an inactivated hepatitis A vaccine administered in two different doses and schedules to 460 healthy volunteers aged 3-18 years. Participants were randomized to two groups to receive either two doses of 720 ELISA Units (EL.U) inactivated hepatitis A per 0.5 ml dose according to a 0, 6-month schedule, or three doses of 360 EL.U according to a 0, 1, 6-month schedule. Transient local injection soreness was the most commonly reported symptom in almost half of both groups with no serious adverse events. One month after the primary course (one dose of 720 EL.U and two doses of 360 EL.U), 99% of 720 EL.U vaccinees had seroconverted, compared with 100% seroconversion in the 360 EL.U group. All vaccinees were seropositive after the booster dose of both vaccines with geometric mean anti-HAV titers of 2,359 and 2,967 mIU/ml in the 720 EL.U and 360 EL.U groups, respectively. The vaccine containing 720 EL.U of antigen per dose offers the advantage of convenience and acceptance of immunization afforded by a two-dose course of vaccination accompanied by a comparable antibody response with that achieved after three doses of vaccine containing 360 EL.U of antigen per dose. PMID- 9876950 TI - Seroprevalence of scrub typhus infection in patients with pyrexia at some malaria clinics in three western provinces of Thailand. AB - In Thailand, the epidemiological data on scrub typhus infection represents only "the tip of an iceberg" especially in malaria clinics where patients come to seek attention because of other febrile illnesses that may have initial clinical signs that are indistinguishable from malaria. The objectives of this study were to determine the prevalence of antibody titers to Orientia tsutsugamushi, and its various strains, among patients at some malaria clinics in three western provinces of Thailand. The sample was represented by 200 patients from 6 malaria clinics in Ratchaburi, Petchaburi and Kanchanaburi provinces between June and November, 1994. Blood specimens were collected with their consent. Immunofluorescent antibody assays (IFA) were used for measuring IgM and IgG antibody titers for scrub typhus infection. The results showed that the prevalence rate for scrub typhus infection (IgM and/or IgG titer > or = 50) was 59.50% (119 cases). The immunofluorescent antibody response to various strains of O. tsutsugamushi showed that co-infections with the Karp, the Gilliam and the Kato strains were the most common (found in 68.10% of cases). Geometric mean antibody titers (GMT) were highest for the Karp strain, followed by the Gilliam then Kato strains. In conclusion, this study indicates that the prevalence rate of scrub typhus is not rare in these areas. PMID- 9876952 TI - Trials in developing countries. AB - Good clinical trials can be carried out in the developing countries but ethical issues concerning the trials are frequently brought up for international debate. The concern has its own merit but can be amended if investigators (local and international) pay serious attention to these criticisms and work out the way that will most benefit the trial participants. Although scientific progress is important, it must come after the rights, the safety and the benefit of the patients. Frequently, the ethical standard is delicately balanced, depending on who looks at it and from what angles they look. Critics, investigators, volunteers, sponsors and regulatory agencies have their own mandates, expectations and limitations, and thus they need to keep an open line of communication in order to benefit all parties involved. PMID- 9876951 TI - Characterization of monoclonal antibodies recognizing alpha and beta subunits of human chorionic gonadotropin hormone. AB - Human chorionic gonadotropin (hCG) hormone is required for maintenance of early pregnancy and is a potential marker in the diagnosis and prognosis of both pregnancy and trophoblastic diseases. Murine hybridomas were generated against purified hCG. Seven hybrid clones secreting antibodies against hCG molecule with IgG1/kappa subclass were established. The indirect ELISA result demonstrated that six MAbs (BEL-1 to BEL-6) recognized hCG in both holo and free beta subunit form with negligible cross-reactivity against a closely related hormone, human luteinizing hormone (hLH). In this fusion, only one MAb (ALC-1) showed a cross reaction with hLH, which designated an alpha subunit specific. The outcome of Western blot ascertained that ALC-1 recognized the conformational epitope on alpha subunit of hCG at Mr 23 kDa band in nonreducing condition (NR). In contrast, epitopes belonging to all MAbs recognized beta subunit of hCG were in linear peptide structure at Mr 34 kDa band (NR) and Mr 26 kDa band (R). These six MAbs were further characterized by using two beta subunit carboxy-terminal synthetic peptides (beta109-119 and beta109-145). Three of them (BEL-1, BEL-3, and BEL-4) recognized only epitope harboring in beta109-145 fragment, the others recognized both types of the synthetic peptide. In order to select the most suitable MAbs specific to beta subunit of hCG for exploiting with ALC-1 in the sandwich-type immunometric assay, competitive ELISA was employed. Six individual MAbs specific to beta subunit of hCG were used to compete with biotinylated ALC-1 to evaluate the proximity of their epitopes on the holo form of hCG molecule. Of all six MAbs, BEL-5 depicted the lowest percent inhibition result, which indicated the bottom-most steric hindrance effect. Consequently, MAb BEL-5 will be the most appropriate antibody to utilize in concert with ALC-1 in place of devising a sandwich-type immunometric assay for measuring holo-hCG level. PMID- 9876953 TI - Report of the European Working Group on Dementia Drug Guidelines Meeting- Brussels, November 1997. AB - A conference was held in Brussels in November 1997 under the auspices of the International Working Group for Harmonization of Dementia Drug Guidelines. The meeting reviewed development of European Dementia Guidelines, with a focus on the roles of academics, industry, the European Commission, the European Parliament, and the European Medicines Evaluation Agency. The objectives of the meeting were to review progress in development of dementia drug guidelines across Europe, with particular reference to harmonization of these efforts with developments in other countries, particularly the United States and Japan. Specific topics covered included issues in developing trials to address disease progression, very mild disease, severe disease, and the non-Alzheimer dementias. Clinical trial designs and the types of measures chosen in these new areas are unlikely to be the same as those used in previous trials for symptomatic relief of mild-to-moderate Alzheimer disease. The assessment tools need to be specific for the purpose of the trial and ongoing research is still needed in this area. Little is known about the natural progression of disease based on the types of measures used in clinical trials. Evidence suggests that rates of decline are variable and nonlinear. Improved collaboration between academics, industry personnel, and regulators, particularly through the sharing of placebo data sets, could allow considerable advances in our understanding of natural disease progression. PMID- 9876954 TI - Alzheimer disease-related activities in China: a report from the International Working Group on the Harmonization of Dementia Drug Guidelines. PMID- 9876955 TI - First International Pharmacoeconomic Conference on Alzheimer's Disease: report and summary. AB - The First International Pharmacoeconomic Conference on Alzheimer's Disease (AD) was held in Amsterdam in July 1998. The meeting was held under the auspices of the International Working Group for Harmonization of Dementia Drug Guidelines (http://dementia.ion.ucl.ac.uk/harmon), bringing together academics, clinicians, purchasers, and representatives from industry. Presentations were given on the methodology of pharmacoeconomic studies in AD, particularly focusing on caregiver burden, quality of life (QOL), and resource utilization. Three economic models of AD were presented based on data from the United States, Canada, and the United Kingdom. In two studies, these data were then used to model the cost effectiveness and effect on cost of treatment with donepezil. Both studies suggested a possible cost advantage for the use of donepezil, when compared with no placebo or treatment, particularly when donepezil is used appropriately in mild-to-moderate AD. These data need to be interpreted with care, as none of the cost or utility information were collected during the clinical trials. Additional data from a 2-year clinical trial of selegiline and vitamin E suggest that cognitive measures may be poor predictors of economic outcome, which is better measured directly. Both economic models of donepezil rely on short-term cognitive data to predict long-term outcome, a methodf that may not be useful in predicting economic savings. The issues facing pharmacoeconomists, researchers, clinicians, and families in the future were addressed in a series of workshops using a method of strategic futuring. The workshops attempted to see 7 years into the future for a range of areas, including consumer and caregiver use of pharmacoeconomic data; early detection and prevention; Japanese perspectives; activities of daily life and what will be daily life activities; caregiver burden; QOL at the end of life; new uses for new information and communication technology in clinical research; and physicians' use of pharmacoeconomic data. A range of exciting futures were predicted, although common themes that arose when considering barriers to achieving these futures included cost, education, political will, confidentiality, privacy, and ethics. The first conference was deemed to have been a success, having attracted more than 160 delegates and many distinguished speaker. A second conference is planned for the year 2000. Over the next 2 years, research needs to be broadened particularly in the methodological areas of resource utilization, QOL, and caregiver burden. Data from clinical trials with relevant economic and QOL outcomes will be needed by purchasers if drug treatments for dementia are to gain widespread use. It is also hoped that the models described at the meeting may become more freely available to politicians, purchasers, clinicians, and caregivers to help them make better decisions about treatment. PMID- 9876956 TI - Clinical trial designs for demonstrating disease-course-altering effects in dementia. AB - Advances in our understanding of the pathogenesis of Alzheimer disease (AD) and vascular dementia (VaD) now permit responsible discussion of therapies that may go beyond relief of cognitive and behavioral symptoms and actually slow progression of disease. The mechanisms of neuronal death and the pathologic role of glia are being elucidated, and epidemiologic studies have suggested potential protective value for anti-inflammatory drugs, estrogen, and free-radical scavengers. However, demonstrating disease-modifying drug effects for progressive conditions such as dementia can be a daunting task, fraught with clinical, statistical, and ethical dilemmas. To evaluate trial designs for demonstrating such effects, the International Working Group on Harmonization of Dementia Drug Guidelines (IWG) conducted a symposium at the Sixth International Congress on Alzheimer's Disease and Related Disorders, held July 1998 in Amsterdam. The presentations at the IWG symposium covered the two basic designs currently being used in clinical trials, survival analysis and staggered-start/withdrawal, in addition to clinical data generated from the National Institute on Aging Alzheimer's Disease Cooperative Study vitamin E/selegiline trial in patients with AD and the phase III clinical studies of propentofylline in patients with AD and VaD. It is hoped that this article will open a dialogue among investigators and regulatory authorities regarding appropriate trial designs to support a regulatory claim for disease-modifying effects. PMID- 9876957 TI - Harmony and consensus: cultural aspects of organization in international science. AB - This article is the beginning of an analysis of cultural processes involved in scientific social organization and change which focuses on the International Working Group for the Harmonization of Dementia Drug Testing Guidelines (IWG). We provide some background for the IWG and for the study of its processes and focus on the construction of consensus and consensus statements of the constituent working groups of the IWG. We delineate a cultural ethos of cooperation that pushes scientists to agree even when they do not and to conceal disagreements. Our observations suggest that the consensus process may serve to inhibit or block the development of new ideas and solutions to problems by keeping conflicts hidden and different viewpoints masked. The masked ideas, although minority ideas, may contain much of importance for subsequent paradigm shifts in approaches to Alzheimer disease and other dementias. In subsequent reports, we will chronicle other aspects of the work of the IWG, including the nature of cross-cultural differences in the drug research structures and processes, governmental regulatory bodies, and the relationship of the pharmaceutical industry to clinical trials research. The present article adds some understanding of the IWG and its mission and contributes to the literature on the cultural construction and social organization of science. PMID- 9876959 TI - A 25-week placebo-controlled study of eptastigmine in patients with Alzheimer disease. AB - The efficacy and safety of eptastigmine in patients with probable Alzheimer disease was evaluated in a double-blind, placebo-controlled, parallel group study. Patients with mild to moderate dementia were randomly assigned to placebo, eptastigmine 10 mg three times a day (t.i.d.), or eptastigmine 15 mg t.i.d. over 25 weeks. The Alzheimer Disease Assessment Cognitive Subscale (ADAS-Cog) and the Clinician's Interview-Based Impression of Change Plus (CIBIC-Plus) were the primary outcome measures for efficacy. Twenty-six centers recruited 320 patients: 106 on placebo, 105 on eptastigmine 10 mg t.i.d., and 109 on eptastigmine 15 mg t.i.d. Six patients on placebo (6%), 18 patients on eptastigmine 10 mg t.i.d. (17%), and 10 patients on eptastigmine 15 mg t.i.d. (9%) discontinued study treatment. The intent-to-treat analysis on 315 patients showed a statistically significant (p=0.047) difference of 2.0 points on ADAS-Cog between the placebo and the eptastigmine 15 mg t.i.d. group at the end of treatment. Patients in the 10 mg t.i.d. group performed better than did placebo-treated patients on the Spontaneous Behavior Interview (SBI) total scores (p=0.015) and on the Activities of Daily Living (ADL, p=0.043) and Behavioral Problems (BP, p=0.028) subscales. The differences in favor of the eptastigmine groups on the CIBIC-Plus did not reach statistical significance. In a post hoc subgroup analysis by staging, the effect size of eptastigmine was found to be greater in the most severely impaired patients (Global Deterioration Scale rating of 4 and 5 at screening) reaching statistical significance in both ADAS-Cog (p=0.007) and CIBIC-Plus (p=0.038). In this patient subgroup (n=222), there was also a significant effect of eptastigmine on SBI (p=0.019). The drug was generally well tolerated, with 8% of patients withdrawing due to adverse events versus 5% on placebo. Adverse events were recorded in 35 patients (33%) on placebo compared with 41 (39%) on eptastigmine 10 mg t.i.d. and 38 (35%) on eptastigmine 15 mg t.i.d. This study shows that eptastigmine doses up to 15 mg t.i.d. for 25 weeks are well tolerated. The drug positively affects cognitive performance of Alzheimer patients. This effect appears greater in more severely impaired patients and also impacts on their behavioral performance. PMID- 9876960 TI - Presentation and stability of noncognitive symptom patterns in patients with Alzheimer disease. AB - The purpose of this study was to investigate noncognitive symptoms in Alzheimer disease to identify symptom patterns and to study stability of such patterns prospectively. Furthermore, variables were examined that could be associated with certain types of symptom patterns or could be predictors of change of these patterns. Forty-eight patients with the clinical diagnosis of probable Alzheimer disease were included in this study and were assessed weekly over a 3-week period. Noncognitive symptoms were rated according to the Behavioral Abnormalities in Alzheimer's Disease Rating Scale and the Dementia Mood Assessment Scale and to a set of items that specifically assess misidentifications. By means of principal component factor analysis different noncognitive symptom patterns were obtained, yielding a four-factor solution. They mapped onto rational domains with respect to clinical experience: depression, apathy, psychotic symptoms/aggression, and misidentifications/agitation. Demographic and clinical variables were not associated with the factor solutions and did not predict change of the factor values. The results demonstrate that in Alzheimer disease there are distinct noncognitive symptom patterns that hold at least short-term prospective stability. None of the examined clinical variables, such as age at entry, the status of the patients (outpatient or inpatient), or dementia severity, exerted substantial influence on the noncognitive symptom patterns. Further investigations should concentrate on the pathological and prognostic correlates of noncognitive symptom patterns in Alzheimer disease. PMID- 9876958 TI - Efficacy of xanomeline in Alzheimer disease: cognitive improvement measured using the Computerized Neuropsychological Test Battery (CNTB). AB - The cognitive efficacy of the M1-selective muscarinic agonist xanomeline in mild to-moderate Alzheimer disease (AD) was measured using the Computerized Neuropsychological Test Battery (CNTB) and the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-cog) in this 17-center, double-blind, placebo controlled study. Three hundred forty-three patients were randomly assigned to receive 25, 50, or 75 mg xanomeline tartrate or placebo three times daily (t.i.d.) for 24 weeks, followed by placebo for 4 weeks in a single-blind washout phase. Cognitive function was assessed at randomization and after 4, 8, 12, 24, and 28 weeks. Three hundred nineteen patients were included in an intent-to-treat (ITT) analysis; 209 completers had evaluable data at week 24. ITT analysis showed a significant (p < or = 0.05) dose-response trend and a significant (p < or = 0.05) between-group comparison favoring 75 mg t.i.d. over placebo for the CNTB summary score but not for the ADAS-cog. In the completer analysis, however, the ADAS-cog showed a significant (p < or = 0.05) dose-response trend and between group comparison, whereas the CNTB Summary Score did not. The ADAS-cog was less sensitive to treatment effects in mildly impaired patients (ADAS-cog < 21) than in moderately impaired patients (ADAS-cog > or = 21), whereas the CNTB was sensitive in the entire study population (mean ADAS-cog = 22.5+/-9.6). Significant (p < or = 0.05) beneficial treatment effects were seen in measures of simple reaction time and delayed verbal recall, which are included in the CNTB but not in the ADAS-cog. During the single-blind placebo washout period, the ADAS cog score of the placebo group worsened dramatically (change of 2.63 points; p < or = 0.001), whereas the CNTB score remained stable (change of 1.04 points; p=0.694). Thus, the CNTB appears to be more objective than the ADAS-cog. PMID- 9876961 TI - Behavioral and psychiatric manifestations in dementia patients in a community: caregiver burden and outcome. AB - The aim of the present study was to document behavioral disturbances in dementia patients in a sample not specifically referred to a clinic. Ninety patients with dementia in a community were studied in relation to the behavioral and psychiatric manifestations as perceived by their caregivers. They were categorized into two subgroups based on severity of the illness, namely mild and moderate-severe, for the purpose of comparison. There were 68 patients with Alzheimer disease, 10 with vascular dementia, and the remaining 12 formed a miscellaneous group.The frequency of the following behaviors in relation to the severity of the dementia were assessed: aggression, physical violence, wandering, incontinence, disinhibition, binge-eating, hallucinations, delusions, and depression. The most common behavioral change was aggression (59%), followed by wandering (27%), delusions (22%), and incontinence (18%). Aggression caused the most distress to the caregiver. There was a higher incidence of wandering, incontinence (p= 0.009), and persecutory delusions (p=0.02) in the moderate severe group. A significantly higher proportion of the moderate-severe group required further care and intervention (p=0.04). This study is probably one of the rare nonclinical surveys on this subject. PMID- 9876962 TI - Frontotemporal behavioral scale. AB - At autopsy, frontotemporal dementia (FTD) account for up to 20% of degenerative dementia cases, although FTDs are underrecognized in memory clinics. FTDs are confused with Alzheimer disease (AD) or vascular dementia (VaD). These misdiagnosis may affect the results of AD pharmacological trials. The first manifestations of FTD are behavioral abnormalities. The aim of this study was to assess a behavioral scale of frontal lobe dysfunction and to determine a behavioral cutoff to diagnose early FTD and distinguish it from AD and VaD. The score of the behavioral frontotemporal lobe dysfunction assessment scale was higher in FTD than in other dementias (p < 0.0001). With a cutoff of 3 points on the scale, FTD patients were diagnosed with a specificity of 95% and sensitivity of 91%. Noncognitive symptoms known to be institutionalization factors could contribute to differences between etiologies of mild dementia. PMID- 9876963 TI - Depressive symptomatology in first-degree family caregivers of Alzheimer disease patients: a cross-ethnic comparison. AB - This study investigated the prevalence of depressive symptoms among White Hispanic (WH) and White non-Hispanic (WNH) first-degree family caregivers. We screened 653 primary caregivers of family members with possible or probable Alzheimer disease who presented at our outpatient memory disorders clinic. Caregiver depression was assessed utilizing the Center for Epidemiologic Studies Depression (CES-D) Scale. Overall, depression (CES-D scores > or = 16) was more common among WH (45%) than among WNH (36%) caregivers (p < 0.05). Elevated CES-D scores among the entire caregiving sample were also linked with being a female spouse (p=0.002), increased level of patient cognitive impairment (p=0.002), and patient psychosis (p=0.002). Risk factors for caregiver depression were identified and compared when the sample was stratified by ethnicity (WH and WNH) and generation (spouses and children). Patient cognitive impairment was a predictor of caregiver depression only among WH spouses and children, whereas patient psychosis was a predictor only among WNH spouses. Female caregiver gender was the most robust risk factor for caregiver depression, being a predictor in all groups except WH children. Implications of this study include the need for increased clinical sensitivity to depression in ethnic minority caregivers, treatment of psychiatric morbidity in dementia caregivers, and respite care for caregivers with high risk for depression. PMID- 9876964 TI - Rate of progression of mental decline in Alzheimer disease: summary of European studies. AB - The aim of this study was to provide an update on the rate of progression in Alzheimer disease. In particular, we conducted a survey on European longitudinal studies that focused on the course of the disease and were performed since 1992. By means of Medline search and review papers and by personal contact with principal investigators, a census of existing studies on natural history of the disease in Europe has been carried out. We present data arising from our search and discuss methodological aspects by comparing data available from U.S. studies on disease progression and implications for clinical trials. PMID- 9876965 TI - Cardiac autonomic dysfunction in patients with Alzheimer disease: possible pathogenetic mechanisms. AB - We studied a possible correlation between autonomic cardiac activity and the level of the red blood cell acetylcholinesterase (AChE) in patients with probable Alzheimer disease (AD). The influence of cholinesterase inhibitor treatment on this autonomic activity was evaluated. Twelve patients satisfying the NINCDS ADRDA criteria of probable AD and 10 healthy controls were studied. Autonomic cardiac activity was evaluated by means of power spectral analysis (PSA) of heart rate variability (HRV) using an autoregressive algorithm on 250 consecutive electrocardiographic R-R intervals. All patients received oral eptastigmine, a new cholinesterase inhibitor, for 1 month. Before treatment, a simultaneous recording of the electrocardiographic and respiratory activities was performed at rest and subsequently during head-up tilt test at 700. Recording was repeated on the last day of treatment. The level of AChE activity during each recording was also evaluated. Spectrum power was calculated in three main frequency bands: high frequency (HF), 0.15-0.4 Hz; low frequency (LF), 0.04-0.15 Hz; very low frequency (VLF), <0.04 Hz. In addition, we calculated the total spectrum power (TSP) and the LF/HF ratio. The TSP and the absolute value of each spectral component were significantly lower in AD patients than in controls. In contrast with controls, AD patients did not show any significant change before treatment in either the LF and HF components or in the LF/HF ratio during the tilt test. However, the modification in the LF component, induced by tilting, showed a significant correlation with the level of AChE activity (p < 0.03). During the tilt test, the treatment caused changes in LF and HF components and in the LF/HF ratio similar to those observed in controls. These results suggest that the presence of autonomic cardiac dysfunction in AD patients might be due to a cholinergic deficit in the peripheral autonomic nervous system. Some aspects of this autonomic dysfunction can be normalized by cholinesterase inhibitor treatment. PMID- 9876966 TI - Lack of association of apolipoprotein E epsilon4 allele dose with cerebral glucose metabolism in Alzheimer disease. AB - Parietal cerebral glucose metabolism is reduced before substantial impairments appeared in subjects carrying the apolipoprotein E (APOE) epsilon4 allele, but the effect of the APOE epsilon4 allele on cerebral metabolism in Alzheimer disease (AD) is still undetermined. To investigate the effect of the APOE epsilon4 allele on cerebral metabolism in AD, we examined regional cerebral glucose metabolism in 83 patients with AD by using 18F-fluorodeoxyglucose and positron emission tomography. Cerebral glucose metabolism in the fronto-parieto temporal association and limbic cortices was significantly decreased in the AD patients compared with 26 age- and sex-matched normal controls. Regional cerebral glucose metabolic rate was not correlated significantly with the number of APOE epsilon4 alleles in any region, which was consistent even after controlling the effects of age, sex, and severity of dementia, and in a subgroup analysis of those aged between 60 and 75. These results supported the view that the APOE epsilon4 allele is not associated with specific deficits in brain metabolism in AD despite evidence of preclinical alterations. PMID- 9876967 TI - The very low density lipoprotein (VLDL) receptor is a genetic susceptibility factor for Alzheimer disease in a European Caucasian population. AB - The epsilon4 allele of the apolipoprotein E gene (APOE) constitutes a major genetic susceptibility factor for Alzheimer disease (AD). Recently, a polymorphic triplet (CGG) in the very low density lipoprotein receptor (VLDL-R) gene, coding for a receptor binding only apoE-containing lipoproteins, was associated with AD in a Japanese population but not in Caucasian American populations. We explored this association and the potential interaction with the APOE polymorphism in a Caucasian sample of sporadic AD and control subjects of similar ages of European origin. The allelic distribution of the VLDL-R polymorphism differed significantly between Japanese and Caucasian populations (p < 0.0001). However, in our population, the presence of at least one VLDL-R 5 repeat allele increased significantly (p < 0.0003) the probability to develop AD after 65 years of age in APOE epsilon4 allele bearers. PMID- 9876968 TI - Tau protein in cerebrospinal fluid is significantly increased at the earliest clinical stage of Alzheimer disease. AB - The concentration of tau protein in cerebrospinal fluid (CSF) was determined in 40 patients with clinically diagnosed probable Alzheimer disease (AD) and in 36 cognitively healthy controls. A significant increase of CSF tau was found in the AD patients, even in 19 subjects with very mild dementia as defined by a Mini Mental State Examination score of 25 and above. Using a cutoff value of 260 pg/mL the sensitivity of elevated tau was 0.89, the specificity was 0.97, and the proportion of correctly allocated cases was 95%. In the AD groups there were no significant associations between CSF tau level and age, age at onset, duration of illness, apolipoprotein E genotype, severity of cognitive impairment, or deficit in regional cerebral blood flow as measured using 99Tm-ethyl cystein dimer single photon emission computed tomography. The findings demonstrate that CSF tau is significantly increased at the earliest clinical stage of AD and shows only minimal overlap with age-matched cognitively healthy controls. This finding suggests that CSF tau could be a biological marker of AD even before dementia has developed. PMID- 9876969 TI - Spatial distribution of diffuse, primitive, and classic amyloid-beta deposits and blood vessels in the upper laminae of the frontal cortex in Alzheimer disease. AB - The spatial distribution of the diffuse, primitive, and classic amyloid-beta deposits was studied in the upper laminae of the superior frontal gyrus in cases of sporadic Alzheimer disease (AD). Amyloid-beta-stained tissue was counterstained with collagen IV to determine whether the spatial distribution of the amyloid-beta deposits along the cortex was related to blood vessels. In all patients, amyloid-beta deposits and blood vessels were aggregated into distinct clusters and in many patients, the clusters were distributed with a regular periodicity along the cortex. The clusters of diffuse and primitive deposits did not coincide with the clusters of blood vessels in most patients. However, the clusters of classic amyloid-beta deposits coincided with those of the large diameter (>10 microm) blood vessels in all patients and with clusters of small diameter (< 10 microm) blood vessels in four patients. The data suggest that, of the amyloid-beta subtypes, the clusters of classic amyloid-beta deposits appear to be the most closely related to blood vessels and especially to the larger diameter, vertically penetrating arterioles in the upper cortical laminae. PMID- 9876970 TI - Intracellular receptors for activated C-kinase in the postmortem human brain: no alteration in Alzheimer disease. AB - Previous studies have identified a 36-kDa protein that has the properties of an intracellular receptor for the activated C-kinase (RACK1). In the present investigation we quantified levels of RACK1 protein in brains from patients with Alzheimer disease (AD) and age-matched controls. Western analysis using a specific antibody showed that the protein levels for RACK1 were not significantly different in AD brains than in controls, indicating that RACK1 is preserved in AD. PMID- 9876971 TI - Suppression of gastric acid secretion: preventing excess gas. PMID- 9876972 TI - Intensive care in developed and developing countries: are comparisons of ICU performance meaningful? PMID- 9876973 TI - Nuclear factor kappaB--a new therapeutic approach? PMID- 9876974 TI - Nuclear factor kappa B: a pivotal role in the systemic inflammatory response syndrome and new target for therapy. AB - NF-kappaB is an important transcription factor complex that appears to play a fundamental role in regulating acute inflammation through activation of the cytokine cascade and production of other pro-inflammatory mediators. There is increasing evidence that NF-kappaB is important in the pathobiology of disease states such as SIRS, MODS and ARDS; therefore, therapeutic interventions aimed at limiting NF-kappaB activation and down-regulating production of inflammatory mediators could prove to be beneficial in decreasing host-derived tissue injury and organ dysfunction. Specific interventions that hold promise for suppressing NF-kappaB activation include the use of antioxidants, inhibition of NIK and the IKK signalsome, treatment with proteasome inhibitors, induction of endotoxin tolerance and, possibly the use of corticosteroids in selected patients. PMID- 9876975 TI - Reliable gastric tonometry after coronary artery surgery: need for acid secretion suppression despite transient failure of acid secretion. AB - OBJECTIVE: To study the need for suppression of gastric acid secretion for reliable intragastric partial pressure of carbon dioxide (PCO2) tonometry by evaluating the effect of an oral dose of sodium bicarbonate before and after administration of the H2-blocker ranitidine to mimic CO2 generation following the buffering of acid by bicarbonate in patients after cardiac surgery. DESIGN: Prospective, open, non-randomized clinical study. SETTING: Cardiothoracic intensive care unit at a university hospital. PATIENTS: 10 patients after elective coronary artery bypass surgery. INTERVENTIONS: An oral dose of 500 mg sodium bicarbonate before and after acid secretion suppression by 100 mg ranitidine as an intravenous bolus given at approximately 3 h after surgery (day 0) and on the first postoperative day (day 1). MEASUREMENTS AND RESULTS: Intragastric PCO2 (iPCO2; tonometry), gastric juice pH (aspirate) and arterial blood gas values were measured. On day 0, the iPCO2 was 25 +/- 5 mmHg before and 31 +/- 5 mmHg after the bicarbonate dose, 29 +/- 5 mmHg after ranitidine infusion, and 31 +/- 5 mmHg after the bicarbonate dose following the ranitidine infusion (NS). On day 1, the basal iPCO2 was 32 +/- 4 mmHg and it increased to 56 +/- 25 mmHg following bicarbonate (p < 0.01). After ranitidine, the iPCO2 was 33 +/- 4 mmHg before and 40 +/- 14 mmHg after bicarbonate (NS). Basal gastric juice pH was > 4 in nine of ten patients on day 0 and > 4 in seven of ten patients on day 1. CONCLUSIONS: Pharmacological suppression of gastric acid secretion is mandatory for reliable iPCO2 tonometry after cardiopulmonary bypass surgery, even when gastric acid secretion is transiently inhibited. In fact, gastric acid secretion was inhibited immediately after surgery, but returned on the first postoperative day in most patients, as judged from the bicarbonate back titration of gastric acid, even when gastric juice pH was relatively high. PMID- 9876976 TI - Intensive care use in a developing country: a comparison between a Tunisian and a French unit. AB - OBJECTIVES: To compare the variations in intensive care (ICU) outcome in relation to variations in resources utilization and costs between a developed and a developing country with different medical and economical conditions. DESIGN AND SETTING: Prospective comparison between a 26-bed French ICU and an 8-bed Tunisian ICU, both in university hospitals. PATIENTS: Four hundred thirty and 534 consecutive admissions, respectively, in the French and Tunisian ICUs. MEASUREMENTS: We prospectively recorded demographic, physiologic, and treatment information for all patients, and collected data on the two ICU structures and facilities. Costs and ICU outcome were compared in the overall population, in three groups of severity indexes and among selected diagnostic groups. RESULTS: Tunisian patients were significantly younger, were in better health previously and were less severely ill at ICU admission (p < 0.01). French patients had a lower overall mortality rate (17.2 vs 22.5%; p < 0.01) and received more treatment (p < 0.01). In the low severity range, the outcome and costs were similar in the two countries. In the highest severity range, Tunisian and French patients had similar mortality rates, while the former received less therapy throughout their ICU stays (p < 0.05). Conversely, in the mid-range of severity, mortality was higher among Tunisian patients, and a difference in management was identified in COPD patients. CONCLUSION: Although the Tunisian ICU might appear more cost-effective than the French one in the highest severity group of patients, most of this difference appeared in relation to shorter lengths of ICU stay, and a poorer efficiency and cost-effectiveness was suggested in the mid range severity group. Differences in economical constraints may partly explain differences in ICU performances. These results indicate where resource allocation could be directed to improve the efficiency of ICU care. PMID- 9876978 TI - Predictors of respiratory function deterioration after transfer of critically ill patients. AB - OBJECTIVES: Critically ill patients are often transferred due to the growing number of diagnostic procedures required to be performed outside the intensive care unit. These transfers have proved to be very critical. The aim of this study was to evaluate predictors for the deterioration of respiratory function in critically ill patients after transfer. DESIGN: Prospective, clinical, observational study. SETTING: 1800-bed university teaching hospital. SUBJECTS: 98 mechanically ventilated patients were investigated during transfer. MEASUREMENT AND MAIN RESULTS: Before transfer, all patients were classified according to the Acute Physiology and Chronic Health Evaluation (APACHE) II score and the Therapeutic Intervention Scoring System (TISS). Haemodynamics and arterial blood gases were measured at 11 different times. Arterial oxygen tension (PaO2), fractional inspired oxygen (FIO2), PaO2/FIO2 ratio, lowest PaO2/FIO2 ratio, minimal PaO2 and maximal FIO2, APACHE II score, TISS before transfer, age and duration of transfer were analysed as potential predictors for deterioration of respiratory function after transfer. Variables were analysed using Classification and Regression Trees and Clustering by Response. In 54 transports (55%) there was a decrease in the PaO2/FIO2 ratio, and a decrease of more than 20% from baseline was noted in 23 of the transferred patients (24%). Age > 43 years and FIO2 > 0.5 were identified as predictors for respiratory deterioration. CONCLUSIONS: Our predictors were able to indicate deterioration after transfer correctly in 20 of 22 patients (91%), combined with a false-positive rate in 17 of 49 (35%). PMID- 9876977 TI - Morphological changes in chest radiographs of patients with acute respiratory distress syndrome (ARDS). AB - OBJECTIVE: To determine whether the quality of infiltrations in chest radiographs can accurately predict the histological extent of fibrotic change in patients with acute respiratory distress syndrome (ARDS). DESIGN: Retrospective clinical investigation. SETTING: Intensive care unit (ICU) of a university teaching hospital. PATIENTS AND METHODS: Of 47 patients treated with extracorporeal membrane oxygenation (ECMO) for severe ARDS over a 5-year period, 23 patients underwent open lung biopsy at thoracotomy for treatment, mostly of pneumothorax. Chest films obtained by portable chest roentgenography preceding the operation were reviewed retrospectively and compared to the histomorphological results of the lung specimen. RESULTS: Chest radiographs displayed mixed alveolar-reticular opacification in 60.2%, alveolar patterns in 22.9% and reticular opacities in 10.5%. In 0.4% there were no infiltrates, 6% could not be evaluated because of insufficient quality. There was no relevant difference between the right and left lungs. Subdividing patients into two groups according to the histological results of either absent or mild (1) or severe (2) lung fibrosis, we found an alveolar haziness in 12.3% in group 1 compared with 28.2% in group 2, while reticular characteristics were identified in 13% and 11%, respectively. CONCLUSIONS: The most common opacity in chest radiographs of patients with severe ARDS treated with ECMO is mixed alveolar-reticular opacification. Severe lung fibrosis is not positively correlated with a reticular radiographic pattern. ECMO does not lead to specific radiological changes in conventional radiograms, contrary to clinical findings that treatment with ECMO might induce pleural or pulmonic haemorrhage, especially in the earlier days when systemic heparinization had to be used instead of the heparin-coated tube-surfacing. PMID- 9876979 TI - Detection of endotracheal tube obstruction by analysis of the expiratory flow signal. AB - OBJECTIVE: Acute obstruction of endotracheal tubes (ETT) increases airway pressure, decreases tidal volume, increases the risk of dynamic hyperinflation by prolonging the duration of passive expiration, and prevents reliable calculation of tracheal pressure. We propose a computer-assisted method for detecting ETT obstruction during controlled mechanical ventilation. The method only requires measurement of the expiratory flow. DESIGN: Computer simulation; prospective study in two cases; retrospective study in one case and in seven patients with the adult respiratory distress syndrome (ARDS). SETTING: Laboratory of the Section of Experimental Anaesthesiology (University of Freiburg); surgical adult intensive care units in a university hospital (University of Basel) and in a university affiliated hospital (Zentralklinikum Augsburg). PATIENTS: 3 patients with partial ETT or bronchial obstructions and 7 ARDS patients. MEASUREMENTS AND RESULTS: Expiratory flow was measured using a pneumotachograph and integrated to obtain expiratory volume. The time-constant of passive expiration (tauE) as a function of expired volume [tauE(V(E)) function] was calculated from the expiratory volume/flow curve. We investigated the tauE(V(E)) function of data obtained from: (1) computer simulation of mechanically ventilated homogeneous and inhomogeneous lungs intubated with ETTs of different sizes; (2) one patient with an artificial ETT obstruction of 7.5 and 25% of the cross-sectional area of the ETT (case 1); (3) one patient with ETT obstruction due to secretions (case 2); (4) one patient with acute bronchial constriction (case 3); (5) seven ARDS patients who showed an increase in airway resistance of more than 2 cm H2O x s/l. It was found that an ETT obstruction caused an increase in tauE in early expiration (at high flow), whereas tauE in late expiration was virtually unchanged. The reason for this is the flow dependency of the increase in ETT resistance produced by ETT obstruction. Unlike ETT obstruction, an increase in pure airway resistance produced an increase in tauE throughout expiration. CONCLUSIONS: An ETT obstruction can be reliably distinguished from an increase in pure airway resistance by a characteristic pattern change in the tauE(V(E)) function, which can be detected easily even by an automated pattern recognition system. PMID- 9876981 TI - Effect of plasma and LPS on respiratory burst of neutrophils in septic patients. AB - OBJECTIVE: To compare the respiratory burst of neutrophils in sepsis and control patients using lipopolysaccharide (LPS), autologous plasma, and a combination of the two. DESIGN: Prospective, consecutive case study. SETTING: A 16-bed intensive care unit (ICU) in a university teaching hospital. INTERVENTIONS: None. PATIENTS: Plasma was obtained from 23 healthy patients scheduled for minor surgery immediately prior to induction of anesthesia (controls) and from 23 ICU patients within 24 h of diagnosis of sepsis or septic shock. MEASUREMENTS AND MAIN RESULTS: Respiratory burst was determined by lucigenin chemiluminescence expressed as mean +/- SEM of peak values of relative light units per neutrophil. There were no significant differences between neutrophils of septic patients and controls for the stimuli saline, phorbol myristate acetate, formyl-methionyl leucyl-phenylalanine, and LPS alone. Septic patients showed a lower respiratory burst than controls (p < 0.05) under the following stimuli: plasma alone (5911 +/ 803 vs 15,397 +/- 3038) and LPS and plasma combined (13,857 +/- 1537 vs 23,026 +/- 2640). However, when stimulated with plasma after priming with LPS, septic patients elicited a higher value than control subjects (11,373 +/- 1758 vs 5987 +/- 1234, p < 0.05). CONCLUSIONS: (1) Some components of the plasma of septic patients may have a profound effect on neutrophil response; (2) plasma as a respiratory burst stimulus differentiates between sepsis and non-sepsis samples better than other common stimuli; (3) precautions must be taken when using plasma together with LPS because of the different response depending on whether LPS priming precedes the plasma stimulus or both are introduced simultaneously and whether septic or nonseptic plasma is used. PMID- 9876982 TI - Point-of-care (POC) measurement of coagulation after cardiac surgery. AB - OBJECTIVE: Two different point-of-care (POC) systems for the monitoring of coagulation variables at the bedside were evaluated with regard to practicability, accuracy and costs. DESIGN: Prospective, descriptive study. SETTING: Single-institutional, clinical investigation on an intensive care unit (ICU) of an urban, university-affiliated hospital. PATIENTS: Eighty cardiac surgery patients were studied postoperatively. INTERVENTIONS: Arterial blood samples were drawn postoperatively on the ICU at different data points. MEASUREMENTS AND RESULTS: Activated partial thromboplastin time (aPTT) and prothrombin time (PT) were measured using two POC systems (Thrombolytic Assessment System [TAS] and CoaguCheck Plus). At the same time coagulation parameters were measured by the central laboratory of the hospital. Measurements were carried out at different data points after cardiac surgery on the ICU. The direct and indirect costs of measuring aPTT/PT were also assessed. Bias analyses revealed good agreement of the POC-based monitoring of aPTT/PT with laboratory based monitoring of coagulation (e. g. aPTT CoaguCheck: bias of -2.8 s with +/- 2 SD [limits of agreement] of +13.7 and -19.1 s). Mean turn-around time (TAT; time from blood sampling until availability of data for the ICU physicians) was significantly longer for the central laboratory-based coagulation monitoring (130 +/- 38 min) than for the two POC systems (aPTT-TAS: 9.6 +/- 2.7 min; aPTT CoaguCheck: 6.5 +/- 1.9 min). Blood sampling at unfavorable times increased the TAT for laboratory-based measurements considerably. The direct costs for measuring aPPT and PT were significantly higher using both POC systems (aPTT-TAS: $4.84; aPTT-CoaguCheck: $4.34) than for the central laboratory ($1.59). Costs for transportation increased the laboratory-based monitoring considerably ($3.77). CONCLUSIONS: Both POC analyzers may reduce the potential for preanalytical errors associated with coagulation measurements at the central laboratory, hasten TAT significantly and may improve patient therapy by reducing inappropriate administration of blood products. PMID- 9876980 TI - Inhaled nitric oxide (NO) for the treatment of early allograft failure after lung transplantation. Munich Lung Transplant Group. AB - OBJECTIVE: Inhalation of high concentrations of nitric oxide (NO) has been shown to improve gas exchange and to reduce pulmonary vascular resistance in individuals with ischemia-reperfusion injury following orthotopic lung transplantation. We assessed the cardiopulmonary effects of low doses of NO in early allograft dysfunction following lung transplantation. DESIGN: Prospective clinical dose-response study. SETTING: Anesthesiological intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: 8 patients following a single or double lung transplantation who had a mean pulmonary arterial pressure (PAP) in excess of 4.7 kPa (35 mmHg) or an arterial oxygen tension/fractional inspired oxygen ratio (PaO2/FIO2) of less than 13.3 kPa (100 mmHg). INTERVENTIONS: Gaseous NO was inhaled in increasing concentrations (1, 4 and 8 parts per million, each for 15 min) via a Siemens Servo 300 ventilator. MEASUREMENTS AND RESULTS: Cardiorespiratory parameters were assessed at baseline, after each concentration of NO, and 15 min after withdrawal of the agent [statistics: median (25th/75th percentiles: Q1/Q3), rANOVA, Dunnett's test, p < 0.05]. Inhaled NO resulted in a significant, reversible, dose-dependent, selective reduction in PAP from 5.5(5.2/6.0) kPa at control to 5.1(4.7/5.6) kPa at 1 ppm, 4.9(4.3/5.3) kPa at 4 ppm, and to 4.7(4.1/5.1) kPa at 8 ppm. PaO2 increased from 12.7(10.4/17.1) to 19.2(12.4/26.0) kPa at 1 ppm NO, to 23.9(4.67/26.7) kPa at 4 ppm NO and to 24.5(11.9/28.7) kPa at 8 ppm NO. All patients responded to NO inhalation (either with PAP or PaO2), all were subject to long-term inhalation (1-19 days). All were successfully weaned from NO and were discharged from the intensive care unit. CONCLUSION: The present study demonstrates that low-dose inhaled NO may be an effective drug for symptomatic treatment of hypoxemia and/or pulmonary hypertension due to allograft dysfunction subsequent to lung transplantation. PMID- 9876983 TI - Do we know the costs of what we prescribe? A study on awareness of the cost of drugs and devices among ICU staff. AB - OBJECTIVE: To evaluate the level of cost awareness of drugs and devices among intensive care unit (ICU) doctors with variable levels of experience (senior intensivists, junior intensivists, residents). DESIGN: Interview-questionnaire. SETTING: ICU of the University of Rome "La Sapienza". PARTICIPANTS: 60 ICU doctors (40 specialists in anaesthesia and intensive care, 20 residents). MEASUREMENTS AND RESULTS: The estimated prices of drugs and devices were compared with the correct prices; responses within a range +/- 20% of the true price were arbitrarily considered correct; all the subgroups of doctors made inaccurate estimates of the prices, showing an absence of any impact of professional experience of cost awareness. CONCLUSION: The doctors in the study showed a high level of inaccurate cost awareness of drugs and devices. PMID- 9876984 TI - Nitric oxide inhibits tissue factor synthesis, expression and activity in human monocytes by prior formation of peroxynitrite. AB - OBJECTIVE: Nitric oxide (NO) has antithrombotic properties by regulating platelet function, whereas direct effects on plasmatic coagulation are rarely described. In sepsis and inflammation, when synthesis of NO, oxygen radicals and toxic metabolites is crucial, the expression of tissue factor (TF) on monocytes stimulated by lipopolysaccharides (LPS) induces intravascular coagulation. This study was performed to examine the influence of NO and the NO-dependent metabolite peroxynitrite on LPS-induced TF expression and activity in human monocytes. DESIGN: Experimental study. SETTING: Laboratory for cell biology. METHODS: Human peripheral blood mononuclear cells were isolated from buffy coats by gradient centrifugation. The NO-releasing compounds SIN1 and NOC18 were used under different conditions. TF antigen was assayed by flow cytometry, and its activity by a clotting assay. TF-mRNA was measured by reverse transcriptase polymerase chain reaction (RT-PCR-ELISA). MEASUREMENTS AND RESULTS: Whereas NOC18, a pure NO donor, had no effect, SIN1, releasing both NO and superoxide (O2 ), reduced TF expression and activity in a dose- and time-dependent manner; superoxide dismutase (SOD) reversed the SIN1-mediated effect. Adding the O2(-) deliberating system hypoxanthin/xanthin oxidase (which had no significant effect per se) to NOC18, or using the NO and O2- reaction product peroxynitrite resulted in a reduction of TF expression. RT-PCR-ELISA indicated upregulation of TF-mRNA by SIN1 with a peak at 500 microM; higher doses had less effect. CONCLUSION: These data demonstrate an influence of NO on LPS-induced TF expression in monocytes by prior formation of peroxynitrite; furthermore, the balance between NO and O2- seems to play a crucial role. PMID- 9876985 TI - Cardiorespiratory, tissue oxygen and hepatic NADH responses to graded hypoxia. AB - OBJECTIVE: To assess cardiorespiratory, tissue oxygen and hepatic nicotine adenine dinucleotide hydride (NADH) responses to graded hypoxia. DESIGN: Prospective, controlled, randomized study. SETTING: University laboratory. ANIMALS AND INTERVENTIONS: 18 anaesthetised Sprague-Dawley rats spontaneously breathing either 21% (controls), 12.5% or 10% inspired oxygen concentrations (6 rats per group). MEASUREMENTS AND RESULTS: All animals in the 21 and 12.5% O2 groups survived the 3-h study period, compared to only 1 in the 10% O2 group. In this latter group, mean arterial pressure and renal blood flow fell immediately with hypoxaemia, whereas aortic blood flow was maintained until the preterminal stages. Critical cellular hypoxia was suggested by an increasingly severe base deficit, an initial rise then a preterminal fall in hepatic NADH intensity and premature death in all but 1 animal. Hepatic NADH fluorescence intensity was unchanged in control animals but showed a progressive rise in the 12.5% O2 group, accompanied by a small though static increase in arterial base deficit. No significant differences were seen in arterial and tissue partial pressure of oxygen between the 12.5 and 10% O2 groups. CONCLUSIONS: This study demonstrates major differences in cardiorespiratory, hepatic NADH and outcome responses to small variations in the degree of hypoxic hypoxia. The fall in NADH fluorescence intensity presages impending death and is likely to reflect failure of cellular metabolic processes. PMID- 9876986 TI - Dopamine clearance in critically ill patients. AB - OBJECTIVE: To examine the validity of the low-dose "renal" dopamine regimen in critically ill patients by investigating the steady-state clearance of dopamine. DESIGN: A prospective clinical study. SETTING: Teaching hospital intensive care unit. PATIENTS: 48 haemodynamically stable patients receiving a dopamine infusion. INTERVENTIONS: Sampling of arterial blood and dopamine infusates. MEASUREMENT AND RESULTS: Plasma and infusate dopamine levels were measured by liquid chromatography with electrochemical detection. Steady-state clearance was determined by dividing the actual infusion rate by the steady-state plasma concentration. Dopamine clearance for the whole group was 46.4 +/- 35.9 ml/kg per min (mean +/- SD), which is significantly lower than 70 +/- 15.2 ml/kg per min reported for elective surgical patients (p = 0.01). Twelve patients with renal dysfunction had significantly lower dopamine clearances (36 +/- 16.6 ml/kg per min) than the remaining 36 patients (61 +/- 38.5 ml/kg per min, p = 0.022). There was a very poor correlation between plasma dopamine level and infusion rate for the group as a whole (r = 0.47), and this worsened (r = 0.31) when only those patients on a "renal" dose of 2-5 microg/kg per min were considered (n = 30). CONCLUSION: Plasma dopamine clearance is lower in critically ill patients and there is a large interindividual variation. It is therefore impossible to predict the plasma level from the infusion rate. Consequently, the concept of a selective renovascular low-dose dopamine infusion is invalid in critically ill patients. PMID- 9876987 TI - Guidelines for the pre-hospital care of patients with severe head injuries. Working Group for Neurosurgical Intensive Care of the European Society of Intensive Care Medicine. PMID- 9876988 TI - Nitric oxide in acute lung injury. PMID- 9876989 TI - Inhaled prostacyclin: from pulmonary hypertension to splanchnic hypoperfusion. PMID- 9876990 TI - A case of crush syndrome occurring during alcohol withdrawal: extended disturbance of consciousness with development of persistent Korsakoff's psychosis. PMID- 9876991 TI - Split lung ventilation in a patient with differential pulmonary hemodynamics. PMID- 9876992 TI - About refusal of care in the ICU. PMID- 9876993 TI - Spine injuries associated with falls from hunting tree stands. AB - Spinal injuries resulting from falls out of tree stands are often associated with concomitant neurologic deficit, prolonged hospitalization, and long-term disability. The purpose of this study was to review the types of spinal injuries that resulted from falls from hunting tree stands. We retrospectively reviewed 27 patients who came to our institution for treatment of spine injuries related to tree-stand accidents between 1981 and 1997. Eleven percent of the falls were alcohol related. Mean height of the fall was 19.6 feet (range, 10 to 35 feet). There were 17 burst fractures, 8 wedge compression fractures, 4 fractures involving the posterior elements, and 1 coronal fracture of the sacral body. Significant neurologic injury occurred in 12 patients (44%). Sixteen patients (59%) had associated injuries. Nine patients (33%) had open reduction, internal fixation, and fusion of their spine fractures. One patient was treated with a halo jacket. The remaining patients were treated in rigid, molded, polypropylene thoracolumbar orthoses or lumbosacral corsets. Accidental falls from tree stands may result in significant spinal fractures often associated with concomitant neurologic injury, extended hospitalization, and permanent disability. Many of these injuries may be prevented through aggressive hunter safety education. PMID- 9876994 TI - Cementless revision of total hip arthroplasty using proximal porous-coated femoral implants. AB - Thirty patients had 32 cementless total hip arthroplasty revisions and were evaluated postoperatively for clinical function (Harris Hip Score) and radiographic evidence of implant stability. Of the 26 femoral components revised, 16 were revised with anatomic long-stem femoral prostheses, and 10 were revised with straight mid-stem-length components. All components were collared and had circumferential proximal fiber-mesh porous coating. Seven of 16 patients had radiographic subsidence after revision with long-stem components (2 to 30 mm); 6 of 10 patients had subsidence after revision with mid-stem femoral components (2 to 25 mm). Of the 13 patients with femoral subsidence, 8 had calcar reconstruction with allograft bone; of the 13 patients without radiographic subsidence, 8 did not require calcar reconstruction. One of 27 fiber-mesh, porous coated acetabular components migrated (30 mm). No components have been removed or revised. Even with circumferential proximal porous coating, femoral implant stability remains unpredictable in total hip arthroplasty revision. PMID- 9876995 TI - Femur fractures in patients with hip arthroplasty: indications for revision arthroplasty. AB - The number of periprosthetic femur fractures has increased due to the increase in the number of patients having total hip arthroplasty. In this study, we define indications for operative treatment in patients with femur fractures after hip arthroplasty. Fifty-three patients with 56 periprosthetic fractures were available for retrospective review of charts, radiographs, and physical examination; 42 fractures were treated with open reduction and internal fixation, 8 had replacement of hip prosthesis, 4 were treated with a retrograde genucephalic nail, and 2 patients were treated conservatively. The choice of treatment depended on the stability of the prosthesis and on the type and location of the fracture. Fifty-two fractures healed primarily. Three patients sustained a refracture, one an additional fracture, and two a deep infection. We recommend treatment with plate fixation for fractures without signs of prosthetic loosening. In fractures with loose implants, revision arthroplasty is required. Distal femoral fractures should be stabilized with a plate or with genucephalic nailing. PMID- 9876996 TI - Salter-Harris type III epiphyseal fractures of the proximal phalanx. AB - Of 823 pediatric hand fractures treated between 1981 through 1996, only 11 (1.3%) were Salter-Harris type III fractures of the proximal phalanx. We review the anatomic basis, etiology, age prevalence, treatment, and results of these epiphyseal injuries. The average age at injury was 14.9 years, with a narrow range from 14 years 2 months to 15 years 11 months. Nine of 11 fractures were displaced and treated with open reduction and internal fixation. At an average follow-up of 21 months, 9 of 11 had full joint motion. The other two had minimal loss of motion, and all had normal function and collateral ligament stability. The displacement and size of the epiphyseal fragment, avulsed and rotated by the collateral ligament, are often underestimated by radiographs. We found that anatomic reduction and fixation yielded good results in all displaced fractures. This restores both the stability of the collateral ligament and a smooth articular surface. PMID- 9876997 TI - Physical abuse of children: a review for orthopedic surgeons. AB - Statistics show that more than half the victims of child abuse have fractures. The orthopedic surgeon will often be the first person to identify a potentially abused child. In a series at Brenner Children's Hospital, 17% of abused children had a fracture as the initial manifestation of child abuse. For protection of the child, it is essential that the orthopedic surgeon recognize the signs of child abuse and make a referral to the appropriate child protection agency. In this review article, we discuss recognition of common types of child abuse and outline appropriate management of child abuse cases. PMID- 9876998 TI - Investigations on the pharmacokinetics of alpha-lipoic acid in healthy volunteers. AB - AIM: The aim of the present study was to examine the pharmacokinetics of racemic alpha-lipoic acid in twelve healthy volunteers following single oral administration of 200 and 600 mg (Thioctacid 200 film tablets). SUBJECTS, MATERIAL AND METHODS: Each film tablet contained 200 mg of alpha-lipoic acid. In addition, an injection solution containing 200 mg of alpha-lipoic acid was administered. Plasma concentrations of alpha-lipoic acid were determined using a validated reversed-phase HPLC method with electrochemical detection having a lower limit of quantitation of 1 ng/ml. The areas under curve (AUC) were 46.82 +/ 21.46 and 157.97 +/- 35.05 microg x min/ml for the oral and intravenous administration of 200 mg, respectively. RESULTS: The AUC following oral administration of 600 mg was 157.83 +/- 35.82 microg x min/ml. The difference in mean t(1/2) for the two oral doses and the i.v. dose (in the range of 25.3-32.7 min) was not statistically significant. CONCLUSION: The lack of a significant difference between values for apparent total plasma clearance for the 200 and 600 mg doses indicates non-saturable kinetics of alpha-lipoic acid in healthy volunteers in this dose range. The absolute bioavailability after the 200 mg dose was 29.1 +/- 10.3%. PMID- 9876999 TI - Integrated pharmacokinetics and pharmacodynamics of the novel calcium sensitizer levosimendan as assessed by systolic time intervals. AB - BACKGROUND: Levosimendan is a new calcium sensitizer, acting calcium-dependently on cardiac troponin C. In the present study pharmacokinetic-pharmacodynamic interrelations of levosimendan were assessed. SUBJECTS AND METHODS: Ten healthy subjects (22-27 years) were given single doses of 2 mg of levosimendan in 4 different formulations: intravenous (i.v.), conventional tablet (CT), conventional capsule (CC), and slow-release tablet (SR) on different days. Systolic time intervals and impedance cardiography were recorded up to 4 hours post drug. Plasma concentrations of levosimendan and its metabolite OR-1855 were analyzed using HPLC. Hysteresis loops were constructed by connecting the effect concentration points in time order. In addition, pharmacokinetic-pharmacodynamic modelling was performed with the i.v. data. RESULTS: The i.v. administration, giving a maximal levosimendan concentration of 180 ng x ml(-1), increased heart rate by 8 beats min(-1) and cardiac output by 18%. It shortened heart rate corrected electromechanical systole QS2i by 23 ms, indicating a fairly strong positive inotropic effect. The conventional oral formulations (giving maximal drug concentrations of about 70-80 ng x ml(-1)) increased heart rate by 4-5 beats min(-1) and cardiac output by 5-8%, while QS2i shortened by 9-13 ms. The SR formulation resulted in low drug concentrations and generally weaker effects than the other formulations. The bioavailability of CT and CC was 83 and 87%, while that of SR was only 31%. QS2i showed counter-clockwise hysteresis after all formulations (p < 0.01). The mean equilibration half-time (ln(2)/k(e0)) after i.v. administration was 9.6 min. Only after SR, OR-1855 was detected in appreciable amounts in plasma, the highest value being 2.2 ng x ml(-1) which occurred 24 hours after drug intake. CONCLUSION: In conclusion, the pharmacokinetic-dynamic behavior of the inotropy index QS2i indicates an equilibration delay of levosimendan, which most probably reflects the time the drug requires to distribute from plasma to its cardiac site of action. The deviant kinetic-dynamic profile of the oral slow-release formulation suggests a different absorption pattern of levosimendan from this formulation. PMID- 9877001 TI - Autoactivation and activation of the cytochrome P450s. AB - OBJECTIVE: In order to reliably predict in vivo pharmacokinetic parameters from in vitro data, we must thoroughly understand the systems we currently use to determine enzyme kinetic parameters. There have been a number of reports of atypical Michaelis-Menten kinetics for cytochrome- (CYP) P4503A mediated metabolism in vitro but little discussion of its clinical relevance. In this manuscript, we examined the scope of CYP autoactivation and confirmed that CYP1A2 demonstrates atypical Michaelis-Menten kinetics in vitro. MATERIALS: Human liver microsomes, baculovirus-expressed CYP1A2, CYP1A2 in the RECO format, and E. coli expressed CYP1A2 were utilized. METHODS: Enzyme kinetics were performed using the various human CYP1A2 sources and ethoxyresorufin O-deethylation as a prototypical biotransformation. The data were fit to various models of enzyme kinetics. In some cases the data best fit the Hill equation, which was used to empirically model allosteric-type autoactivation kinetics. RESULTS: RECO CYP1A2 and E. coli expressed CYPIA2 both demonstrated autoactivation kinetics for ethoxyresorufin O deethylation. When the data were fit to the Hill equation, n (the slope factor) was found to be 1.4 and 1.8 for RECO and E. coli expressed CYP1A2, respectively. Human liver microsomal and insect expressed sources of CYP1A2 illustrated classical Michaelis-Menten kinetics for the O-deethylation of ethoxyresorufin. CONCLUSION: Data generated in the current study and previous work suggest many CYPs, not only CYP3A, appear to behave as allosteric enzymes. We would argue that this is not necessarily a classical allosteric mechanism because n is frequently a non-integer. This autoactivation appears to be a function of several factors including substrate physicochemical characteristics, specific interactions of the substrates (activators) with the enzyme active site, and presence of other enzyme modulators. These factors interact to increase the catalytic activity of CYP and thus the complexity of predicting enzyme kinetic parameters or drug interactions. PMID- 9877000 TI - Bioavailability of repaglinide, a novel antidiabetic agent, administered orally in tablet or solution form or intravenously in healthy male volunteers. AB - OBJECTIVE: Repaglinide is a novel prandial glucose regulator (PGR) for the treatment of type 2 diabetes. In order to investigate subject variability following oral administration of repaglinide, and to determine the relative and absolute bioavailabilities of repaglinide following oral or intravenous administration, two single-centre, open-label, randomized, crossover clinical studies were conducted. SUBJECTS AND METHODS: Study 1 was conducted in 24 healthy male subjects (aged 18 to 49 years), who received repaglinide 2 mg, as either tablet or oral solution, twice each on 4 separate occasions at least 7 days apart. Study 2 was conducted in 12 healthy male subjects (aged 18 to 45 years), who received repaglinide 2 mg, either as a tablet or as an intravenous infusion over 15 minutes, once each on 2 separate occasions, with a washout period of 7-10 days. RESULTS: In study 1 there was no significant difference between administration of repaglinide 2 mg, in either tablet or oral solution form with regard to intrasubject variation in AUC and Cmax. However, the intrasubject variation in t(max) and mean residence time (MRT) was significantly (p = 0.001) larger for the tablets than for the oral solution. Intersubject variation (CV) in AUC ranged from 44.7% to 62.1% after oral administration. The relative bioavailability of repaglinide (AUC(tablet)/AUC(oral solution)) was 110% (95% CI, 103%-117%). In study 2 the absolute bioavailability of repaglinide administered as a tablet was 62.5% (95% CI, 49.2%-79.5%) relative to an intravenous infusion of the same dose. CONCLUSION: There was no evidence from either study that the tablet formulation led to greater variation in serum profiles of repaglinide. It was concluded that repaglinide is rapidly absorbed and eliminated in healthy subjects when administered orally or intravenously under fasting conditions, and that the total availability of repaglinide is similar in the tablet and oral solution formulations, though that the rate of absorption is slower for the tablet formulation. PMID- 9877002 TI - Pharmacokinetic and pharmacodynamic aspects of salmeterol therapy. AB - BACKGROUND: Salmeterol xinafoate is a highly selective beta2-adrenoceptor for the maintenance treatment of asthma in adults and children. OBJECTIVE: To review the pharmacokinetics, clinical pharmacology, and therapeutic properties of a recently introduced, long acting antiasthmatic drug. METHODS: Recent English-language publications were selected using Medline as database. RESULTS: Salmeterol's pharmacokinetics, clinical pharmacology, and therapeutic properties are reviewed and aspects related to salmeterol's unusual duration of action, its high potency, beta2-selectivity, possible antiinflammatory actions, its interaction with other drugs, low systemic adverse effects, dosage, and administration are also discussed. CONCLUSION: Salmeterol is a safe long-acting beta2-agonist very useful for maintenance treatment of asthma. PMID- 9877003 TI - Comparative bioavailability of two sertraline tablet formulations in healthy human volunteers after a single dose administration. AB - OBJECTIVE: To compare the bioavailability of 2 sertraline tablets formulations (Tolrest from Laboratorios Biosintetica, and Zoloft from Laboratorios Pfizer, Brazil) in 24 healthy volunteers of both sexes (12 male and 12 female) who received a single 50 mg dose of each sertraline formulation. MATERIAL AND METHODS: The study was conducted open with randomized two-period crossover design and a 14-day washout period. Plasma samples were obtained over a 96-hour interval and sertraline concentrations were analyzed by combined reversed phase liquid chromatography and tandem mass spectrometry (LC-MS-MS) with positive ion electrospray ionization using selected ion monitoring method. From the plasma sertraline concentration vs. time curves the following pharmacokinetic parameters were obtained: AUC(0-96h), AUC(0-infinity), Cmax, Cmax/AUC(0-96h), Tmax, ke, and t(1/2). RESULTS: Pharmacokinetic parameters presented normal distribution according to Probit' s plot and Kolmogorov Smirnov's test, and the variance of AUC(0-96h), AUC(0-infinity) or Cmax were homoscedastic. Geometric mean Tolrest/Zoloft individual percent ratio was 95.22% for AUC(0-96h), 99.87% for Cmax, 100.4% for AUC(0-infinity), 103.6% for Ke, 96.0% for t(1/2) and 93.7% for Tmax. CONCLUSION: Since the 90% CI for both Cmax and AUC(0-96h) mean ratio were within the 80-125% interval proposed by the Food and Drug Administration, it was concluded that Tolrest was bioequivalent to Zolof for both extent and rate of absorption in a single dose administration. PMID- 9877004 TI - Discriminative stimulus effects of S(-)-methcathinone (CAT): a potent stimulant drug of abuse. AB - Methcathinone ("CAT") is a CNS stimulant that is a very significant drug of abuse in the former Soviet Union. It has also appeared on the clandestine market in the United States and has been recently classified as a Schedule I substance. In the present study, S(-)-methcathinone [S(-)-CAT, 0.50 mg/kg, IP] was employed as the training drug in a two-lever drug discrimination task in rats. Once established, the S(-)-CAT stimulus was shown to have a rapid onset to action (within 5 min) and a duration of effect of approximately 60-90 min. In tests of stimulus generalization (substitution), the S(-)-CAT (ED50 = 0.11 mg/kg) stimulus generalized to S(+)-methamphetamine (ED5 = 0.17 mg/kg), S(-)-cathinone (ED50 = 0.19 mg/kg), S(+)-amphetamine (ED50 = 0.23 mg/kg), aminorex (ED50 = 0.27 mg/kg), (+/-)-CAT (ED50 = 0.25 mg/kg), (+/-)-cathinone (ED50 = 0.41 mg/kg), R(+)-CAT (ED50 = 0.43 mg/kg), cis-4-methylaminorex (ED50 = 0.49 mg/kg), methylphenidate (ED50 = 0.83 mg/kg), and cocaine (ED50= 1.47 mg/kg). S(-)-CAT-stimulus generalization did not occur to fenfluramine, a structurally related nonstimulant anorectic. Lastly, haloperidol (AD50 = 0.18 mg/kg), a dopamine receptor antagonist, potently antagonized the S(-)-CAT stimulus. It is concluded that S(-) methcathinone is a very potent CNS stimulant, which appears to produce its stimulus effect, at least in part, via a dopaminergic mechanism. PMID- 9877005 TI - Behavioral effects of nicotine, amphetamine and cocaine under a fixed-interval schedule of food reinforcement in rats chronically exposed to caffeine. AB - Epidemiological surveys demonstrate that caffeine, the main psychoactive ingredient of coffee, is a positive correlate in drug abuse. To characterize the behavioral nature of caffeine interactions with other psychomotor stimulants, we examined the effects of chronic caffeine exposure on the behavioral responses to nicotine, amphetamine, cocaine, the selective D1 agonist SKF-82958 and the selective D2 receptor agonist NPA, in rats responding under a fixed interval (FI) schedule of food reinforcement. Following stabilization of rates and temporal patterns of responding (mathematically expressed as quarter-life values, QL), twenty-one Sprague-Dawley rats responding under a 5-min FI schedule of food reinforcement were divided into two groups; one (twelve rats) maintained on tap water (control) and the other (nine rats) on caffeine (3 mg/ml added to the drinking water). Following the substitution of caffeine solution for tap water, behavior was temporarily disrupted as evidenced by decreases in responding and QL values which reached a maximum after 72 h (rate 60% and QL 30% below baseline levels). Rats developed complete tolerance to these effects of caffeine over 5 days of caffeine exposure. After response rate and QL values stabilized, effects of drugs were evaluated. Nicotine (0.01-1.0 mg/kg; SC), amphetamine (0.1-5.6; IP), and cocaine (1.0-17; IP) each produced biphasic dose-dependent changes in response rate with maximum increases in response rate following intermediate doses and decreases in response rates following higher doses. The increase in rates of responding produced by amphetamine or cocaine (but not nicotine) were greater (P<0.05) in caffeine-drinking than in water-drinking rats. Both SKF-82958 (0.001-0.3 mg/kg; IP) and NPA (0.0001-0.1; IP) produced only dose-dependent decreases in rates of responding. Caffeine-drinking rats were less sensitive to the rate-depressant effects of SKF-82958 (P<0.05) than water-drinking rats. However, similar changes (P>0.05) were produced by NPA in both groups. Except for amphetamine, the remaining drugs produced similar (P>0.05) dose-dependent decreases in QL values in water- and caffeine-drinking rats. Amphetamine produced smaller decreases in QL values in caffeine-drinking rats than in water-drinking rats (P<0.05). Chronic exposure to caffeine produced complete insurmountable tolerance to the response-rate increasing (stimulant) effects of acute caffeine (3.0-17 mg/kg; IP) in caffeine-drinking rats. In conclusion, our study revealed that chronic caffeine exposure potentiates the behavioral response to amphetamine and cocaine but not to that of nicotine in rats responding under a FI schedule of food reinforcement. Thus, it is likely that these effects are mediated through different pharmacological mechanisms. PMID- 9877006 TI - Facilitation of stimulus detection performance of rats with d-amphetamine: a function of dose and level of training. AB - Conditions under which amphetamine may facilitate stimulus detection task choice performance in rats were investigated. Rats (n=15) were trained in a two-choice, light-detection task to three successively more stringent criterion levels of task training (minimal, intermediate, and extended) and then tested after administration of saline, 0.25, 0.50, and 0.75 mg/kg d-amphetamine (AMP). For each training level, baseline levels of choice accuracy were maintained at approximately 82% by manipulating the animals' cue duration. No aspect of performance was enhanced by any dose of AMP after minimal criteria training, and there was a dose-dependent decrease in the number of trials completed. After the intermediate level of training, the 0.25 mg/kg dose of AMP reliably increased choice accuracy, there was no reliable change in choice reaction time, and there was a dose-dependent decrease in the number of trials completed. After the extended training, the 0.25 mg/kg dose of AMP reliably increased choice response accuracy, the 0.25 and 0.50 mg/kg doses of AMP reliably decreased choice reaction time, and there was no reliable change in the number of trials completed at any dose of AMP. These results support the contention that psychostimulants can facilitate the choice performance of rats in stimulus detection tasks if an appropriately low dose is used and the animal's behavior is strongly controlled by the stimulus-reinforcement contingencies of the task. PMID- 9877007 TI - Effect of histamine H2 and H3 receptor modulation in the septum on post-training memory processing. AB - We compared the effects of modulating the postsynaptic histamine receptor subtype 2 (H2) and inhibitory presynaptic autoreceptor subtype 3 (H3) on memory processing in the septum. Mice were partially trained on footshock avoidance in a T-maze. Immediately after training, saline or a drug solution was infused into the septum. One week later, retention was tested by continuing training until the mice made five avoidance responses in six consecutive trials. The results indicate that dimaprit, an H2 agonist, facilitated retention (25 and 50 pg) with a U-shaped dose-response curve typical of drugs acting at postsynaptic receptors. Cimetidine, an H2 antagonist, impaired retention (15-50 ng). The H3 agonist. imetit, impaired retention (25-200 ng), while the H3 antagonist, thioperamide, facilitated retention (10-400 ng). An unusual feature of the dose-response curve for thioperamide was that it did not appeal to yield a U-shaped curve as occurs with drugs acting postsynaptically, but facilitated retention to approximately the same degree from 50 to 400 ng. As histamine neurons project to various limbic system structures involved in memory processing, it may play an important role in regulating the activity of structures such as the septum, hippocampus and amygdala. PMID- 9877008 TI - Oxotremorine suppresses thalamocortical oscillations via thalamic muscarinic acetylcholine receptors. AB - We investigated whether the local intrathalamic infusion of a muscarinic acetylcholine receptor agonist (oxotremorine) at either the reticular nucleus of thalamus (NRT) or the ventroposteromedial nucleus of thalamus (VPM) suppresses thalamocortically generated neocortical high-voltage spindles (HVSs). In addition, we studied whether the intracerebroventricular (ICV) infusion of a selective muscarinic M2 acetylcholine receptor antagonist (methoctramine) could block the suppression of HVSs induced by either systemic (IP) administration of an anticholinesterase drug [tetrahydroaminoacridine (THA)] or ICV infusion of oxotremorine in rats. Intrathalamic administration of oxotremorine at 3 and 15 microg in the NRT, and at 15 microg in the VPM suppressed HVSs. ICV oxotremorine at 30 and 100 microg and IP THA at 3 mg/kg decreased HVSs. ICV methoctramine at 100 microg increased HVSs and completely blocked the decrease in HVSs produced by oxotremorine 100 microg and THA 3 mg/kg. The results suggest that activation of muscarinic M2 acetylcholine receptors in thalamic nuclei (NRT and VPM) can suppress thalamocortical oscillations and that ICV or systemically administered drugs that activate either directly (oxotremorine and methoctramine) or indirectly (THA) the muscarinic M2 acetylcholine receptors may modulate neocortical HVSs via the thalamus. PMID- 9877009 TI - Gepirone and 1-(2-pyrimidinyl)-piperazine in vitro: human cytochromes mediating transformation and cytochrome inhibitory effects. AB - Biotransformation of gepirone to its principal metabolite, 1-(2-pyrimidinyl) piperazine (1-PP), was studied in human liver microsomes and in microsomes from cDNA-transfected human lymphoblastoid cells. Formation of 1-PP from gepirone in liver microsomes proceeded with a mean apparent Km ranging from 335 to 677 microM. Coincubation with 1 microM ketoconazole reduced reaction velocity to less than 5% of control values at a gepirone concentration of 250 microM. Three other metabolites, presumed to be hydroxylated products, were also formed from gepirone. Formation of all three products was reduced to approximately 20% of control values by 1 microM ketoconazole; quinidine at 1 microM produced a small reduction in formation (91-94% of control) of two of the metabolites. 1-PP was formed from gepirone exclusively by pure P450-3A4 with a Km of 849 microM; Km values for the other metabolites were 245, 240, and 415 microM. Two of the products were also formed by P450-2D6. The results indicate that 3A4 is the principal cytochrome mediating 1-PP formation, as well as formation of the other metabolites. The properties of gepirone and 1-PP themselves as cytochrome inhibitors were tested in human liver microsomes using index reactions representing activity of P450-1A2, -2C9, -2C19, -2D6, -2E1 and -3A. Gepirone and 1-PP produced negligible inhibition of all these reactions. Thus gepirone at therapeutic doses in humans has a low likelihood of inhibiting P450-mediated drug metabolism involving these cytochromes. PMID- 9877010 TI - Effects of intra-amygdala R(+) 7-OH-DPAT on intra-accumbens d-amphetamine associated learning. I. Pavlovian conditioning. AB - We have previously obtained evidence that the mesoamygdaloid dopamine projection modulates the acquisition of a conditioned response (CR) elicited by presentation of a conditioned stimulus (CS) predicting the availability of a natural (sucrose) reward. This property was found to be dependent upon D3, but not D1 or D2, dopamine receptor activation. The aim of the present study was to determine whether the mesoamygdaloid dopamine projection is similarly involved in the acquisition of a drug-associated CR. Thus, two groups of rats with guide cannulae aimed at the nucleus accumbens and amygdala were trained using a Pavlovian conditioning procedure in which an initially neutral CS was paired with a computer-controlled, bilateral intraaccumbens infusion of d-amphetamine (the unconditioned stimulus: US). Conditioning sessions were conducted in standard operant chambers, with each session consisting of a single CS-US trial. For one group of rats, CS presentation was positively correlated with the drug US (Paired group), while for the second group CS and US presentations were negatively correlated (Unpaired group). During training, locomotor activity was recorded and was utilised as the measure both of the unconditioned (UR) and conditioned response (CR). A within-subjects design was utilised to investigate the effect of post-session bilateral intraamygdala administration of R(+) 7-OH-DPAT on the development of the drug-associated CR. Hence, both Paired and Unpaired groups were exposed to two different CSs which were presented on alternate sessions. Post-session bilateral intra-amygdala administration of R(+) 7-OH-DPAT (10 nmol) followed sessions in which one CS was presented, while intra-amygdala vehicle followed sessions in which the alternate CS was presented. The development of a CR occurred only in the presence of a CS that had been positively correlated with presentation of the drug US. Post-session, intra-amygdala administration of R(+) 7-OH-DPAT enhanced the acquisition of this CR. However, R(+) 7-OH-DPAT was without effect upon the unconditioned response to intra-accumbens d-amphetamine. Our previous data indicate a comparable effect of R(+) 7-OH-DPAT on conditioning to a CS associated with a non-drug, natural reward. Therefore, taken together, these findings suggest that D3 dopamine receptors within the amygdala modulate specifically the acquisition of Pavlovian conditioned responses, regardless of whether drug or natural rewards constitute the US. PMID- 9877011 TI - Effects of intra-amygdala R(+) 7-OH-DPAT on intra-accumbens d-amphetamine associated learning. II. Instrumental conditioning. AB - Rats were trained to associate an initially neutral conditioned stimulus (CS) with a response-independent, intra-accumbens infusion of d-amphetamine (the unconditioned stimulus; US). Elsewhere, we have reported that as a result of this training, presentations of the CS alone elicited a conditioned response consisting of increased locomotor activity and that acquisition of this conditioned response was enhanced by post-session, intra-amygdala infusion of the dopamine D3 receptor preferring agonist, R(+) 7-OH-DPAT. Here, in this same group of animals, we have examined the conditioned rewarding properties of the drug associated CS by determining its ability to support the acquisition of a novel instrumental response in the absence of drug reward. Thus, rats were presented with two novel levers. Presentation of the drug-associated CS was made contingent upon depression of one of the levers (CR lever), while responding upon the other lever (NCR lever) had no programmed consequences. Preferential responding upon the lever delivering the drug-associated CS was observed despite a 6-week interval between CS-US training and the conditioned reward test. Intra-accumbens administration of d-amphetamine (0-20 microg) increased the control over behaviour exerted by the CS, increasing CR. but not NCR lever responding. In contrast, rats that received three pairings of an intra-accumbens infusion of d amphetamine in combination with intra-amygdala infusion of R(+) 7-OH-DPAT, 3 weeks prior to testing, displayed similar rates of response upon both levers and were insensitive to the potentiation of responding for conditioned reward following intra-accumbens d-amphetamine. However, intra-accumbens d-amphetamine stimulated locomotor activity in a similar, dose-related manner in both groups. In this way, rats that had received intra-accumbens infusion of d-amphetamine in combination with intra-amygdala infusion of R(+) 7-OH-DPAT appeared exactly like control group rats, for which the CS had been paired with intra-accumbens d amphetamine on a negative basis only. A locomotor activity test indicated that one behavioural consequence of intra-amygdala administration of R(+) 7-OH-DPAT was the reduction of the unconditioned locomotor response resulting from intra accumbens administration of d-amphetamine. Hence, the present data demonstrate that the conditioned rewarding properties of a drug-associated CS are specific to the CS-US association and are relatively insensitive to decay over time. However, the rewarding properties of a drug-associated CS were selectively abolished following activation of amygdala D3 receptors during presentation of the drug reward. Potential explanations for this effect are discussed, including the possibility that intra-amygdala R(+) 7-OH-DPAT reduced the incentive value of the US. PMID- 9877012 TI - The pharmacology of impulsive behaviour in rats IV: the effects of selective serotonergic agents on a paced fixed consecutive number schedule. AB - An impulsive cognitive style may affect behaviour in several different ways, including rapid decision making, intolerance of the delay of reward and a tendency to terminate chains of responses prematurely. It has been proposed to measure the last of these in rats using fixed consecutive number (FCN) schedules. The present study uses a modified version of the FCN procedure in which responding was paced by retracting the response lever for short periods between presses. In this way, the experimenter can control the maximum rate of responding. The procedure was made up of two components. In both, the schedule requirement was FCN 8, but in the Fast component lever presses were spaced by a minimum of 2.5 s and in the Slow component by a minimum of 5 s. Alterations in impulsivity were inferred from changes in the mean chain length and the distribution of chain lengths. The 5-HT1A agonist, 8-OH-DPAT (0.03-0.3 mg/kg), increased chain lengths within a narrow dose range, whereas the 5-HT1A antagonist, WAY 100 635 (0.03-0.3 mg/kg), reduced chain lengths. The 5-HT2 agonist, DOI (0.1-1.0 mg/kg), markedly reduced chain lengths, whereas the 5-HT2 antagonist, ritanserin (0.03-0.3 mg/kg), had no effect. The 5-HT1A/1b agonist, RU 24969 (0.03-0.3 mg/kg), reduced chain lengths. The 5-HT releaser, p chloramphetamine (0.1-1.0 mg/kg), had a weak, biphasic effect, slightly reducing the number of short chains at the lowest dose tested and slightly increasing this number at the highest dose. Other drugs tested, citalopram (1.0-10.0 mg/kg), metergoline (0.3-3.0 mg/kg) and MDL-72222 (0.1-3.0 mg/kg), had no significant effects. These results suggest that stimulation of 5-HT1A receptors reduces impulsivity, whereas stimulation of 5-HT2 receptors increases it. These data are in agreement with previous results using the DRL-72 schedule, and indicate that there is no simple role for serotonin in the control of impulsivity. PMID- 9877013 TI - Acquisition, maintenance and reinstatement of intravenous cocaine self administration under a second-order schedule of reinforcement in rats: effects of conditioned cues and continuous access to cocaine. AB - Second order schedules of IV cocaine reinforcement in rats provide a reliable method for evaluating the effects of conditioned stimuli on cocaine-seeking behaviour, and for measuring the motivational aspects of cocaine reinforcement. In the procedure established here, each infusion of cocaine (0.25 mg/infusion) was initially made contingent on a lever press and was paired with a 20-s light conditioned stimulus (CS). When rats acquired stable rates of cocaine self administration, the response requirement for cocaine was increased progressively to a second-order schedule of the type FI15 min(FR10:S), whereby the IV cocaine infusion was self-administered following the completion of the first FR10 responses (and CS presentation) after a 15-min fixed interval (FI) had elapsed. Evaluation of the animals' responding during the first, drug-free interval of each daily session provided a measure of cocaine-seeking behaviour, independent of other pharmacological effects of the self-administered drug. Thus, a dose response study (dose range: 0.083, 0.25 and 0.50 mg/infusion) revealed that responding under this schedule during the initial, drug-free interval changed monotonically with dose, whereas an inverse relationship between cocaine dose and response level tended to appear during the rest of the session, after rats had self-administered the drug. Responding under this schedule was also shown to occur under the control of the CS, which had acquired conditioned reinforcing properties. Thus, a decrease in responding and an increase in the latency to initiate responding followed the omission of the CS for 3 consecutive days. In addition, extinction of cocaine-seeking behaviour was slower when contingent CS presentations occurred compared to extinction when the CS was not present. Furthermore, the reinstatement of responding for cocaine, which followed a brief period of non-contingent CS presentations, was retarded when this conditioned reinforcer had been extinguished together with cocaine. Finally, cocaine-seeking behaviour decreased markedly for the first 6 h that followed a 12-h period of continuous access to cocaine, when compared to responding 6 h after a 90-min session of limited access to the drug. Responding subsequently increased to baseline levels within 72 h. These results emphasise the utility of second-order schedules for studying drug-seeking behaviour and the importance of drug associated cues in maintaining such responding for cocaine. PMID- 9877014 TI - An investigation of the effects of benzodiazepine receptor ligands and of scopolamine on conceptual priming. AB - Scopolamine and lorazepam both produce anterograde impairments of explicit memory but only lorazepam impairs implicit memory as assessed by perceptual priming tasks. The main aim of the two experiments reported in this article was to determine the effects of these drugs on conceptual priming. Experiment 1 compared the effects of lorazepam (1,2 mg PO) with scopolamine (0.3,0.6 mg SC) and placebo in a study with 60 healthy volunteers. Experiment 2 compared the separate and combined effects of lorazepam (2 mg PO) and flumazenil (2 mg IV) with placebo in a study with 48 healthy volunteers. We found that conceptual priming in category generation tasks was intact following lorazepam in both studies. This preservation of conceptual priming contrasted with lorazepam-induced impairments on explicit memory tasks. In conjunction with previous findings, these results are interpreted as providing further support for the notion that conceptual and perceptual priming are subserved by distinct memory systems, one based on the operations of semantic memory, the other possibly based on a perceptual representation system. That lorazepam impairs perceptual but not conceptual priming suggests that the neurochemical substrates of the two kinds of priming are distinct. PMID- 9877015 TI - Post-session sulpiride infusions within the perifornical region of the lateral hypothalamus enhance consolidation of associative learning. AB - Whilst neurons within the lateral hypothalamus are well known to be responsive to the presentation of previously learned associative stimuli, the consolidation of a Pavlovian association is thought to depend in large part upon other brain regions, including the amygdala. The present study addressed this assumption directly, by examining the effect of post-session infusions of sulpiride within the lateral hypothalamus upon the acquisition of a conditioned approach response in an appetitive differential conditioning task. Subjects were exposed to an initially neutral stimulus (CS+), which immediately preceded the availability of a 10% sucrose reward (US). A second, control stimulus (CS ) was also presented. but never in close temporal proximity to the US. The number and duration of alcove approaches were recorded. Immediately following each training session, subjects were infused bilaterally with sulpiride (0, 0.5, 5 microg) in the vicinity of the perifornical region of the lateral hypothalamus. Sulpiride dose dependently enhanced the rate of acquisition of a conditioned approach response to presentation of the CS+, but was without affect upon approach behaviour during CS(-) or US presentations. Thus, 0.5 microg sulpiride facilitated at an early stage (session 2 onwards) the number of alcove approaches to the CS+, while 5 microg sulpiride enhanced to a greater extent the duration of conditioned approach, particularly during later sessions. A subsequent locomotor test using 0.5 mg/kg d-amphetamine indicated that repeated infusions of the higher dose sulpiride (5 microg), but not the lower dose (0.5 microg), resulted in behavioural sensitisation to administration of the psychomotor stimulant. Acquisition of a novel conditioned instrumental response was not affected by previous exposure to sulpiride. These data suggest that dopamine-sensitive neurons within the lateral hypothalamus may play a significant role in the acquisition of appetitive Pavlovian associations. PMID- 9877016 TI - Sumatriptan (5-HT1D receptor agonist) does not exacerbate symptoms in obsessive compulsive disorder. AB - The non-selective serotonin (5-HT) receptor agonist meta-chlorophenylpiperazine (mCPP) has been reported to elicit symptoms in patients with obsessive compulsive disorder (OCD). MK-212, another non-selective 5-HT receptor agonist, does not seem to induce obsessive compulsive symptoms in OCD patients. The major pharmacological difference between mCPP and MK-212 is their affinity for the 5 HT(ID) receptor. The aim of this study was to explore the role of the 5-HT(ID) receptor in the pathophysiology of OCD, by using a challenge paradigm with the selective 5-HT(ID) receptor agonist sumatriptan (Imigran). A randomized, double blind, placebo-controlled crossover challenge with sumatriptan (100 mg PO) was performed in 15 OCD patients. Neither the obsessive compulsive symptoms nor mood or anxiety symptoms changed significantly following sumatriptan administration as compared to placebo. Sumatriptan did induce a significant increase in plasma growth hormone (GH) levels. In the present study, no indication were found for the role of the 5-HT(ID) receptor in the pathophysiology of OCD. It should be noted, however, that sumatriptan does not readily pass the blood-brain barrier. Selective 5-HT(ID) receptors with better brain penetrating properties may shed more light on the role of this 5-HT receptor subtype in OCD. PMID- 9877017 TI - The nitric oxide synthesis inhibitor nitro-L-arginine (L-NNA) attenuates nicotine abstinence syndrome in the rat. AB - Nitric oxide synthesis contributes to opiate tolerance and dependence. Nicotine dependence and abstinence syndrome in the rat appear to involve opiate mechanisms. Therefore, it was postulated that nitric oxide synthase (NOS) activity might be essential for the expression of nicotine abstinence syndrome. Twenty-one rats were rendered dependent by SC infusion of 9 mg/kg per day nicotine tartrate via Alzet osmotic minipump. Rats were pretreated SC with vehicle alone, or with 18 or 30 mg/kg of the NOS inhibitor L-NNA (nitro-L arginine). Thirty minutes later, rats were challenged by 1 mg/kg of the nicotinic antagonist mecamylamine SC and observed for 30 additional minutes. Rats pretreated with vehicle displayed a total of 68.7+/-8.0 mecamylamine-precipitated abstinence signs (mean+/-SEM), while those receiving 18 or 30 mg/kg L-NNA had 12.7+/-2.0 and 5.1+/-1.7 signs, respectively. All three groups differed significantly from one another according to Dunn's post-hoc procedure. Rats pretreated with L-NNA combined with an excess of the NOS substrate L-arginine had significantly more mecamylamine-precipitated abstinence signs than rats receiving L-NNA combined with D-arginine. Also, D-NNA, which does not selectively bind to NOS, was significantly less effective than L-NNA in preventing mecamylamine precipitated abstinence syndrome. Additional studies determined the effect of L NNA on spontaneous nicotine abstinence syndrome. Rats were assessed for abstinence signs at 17 and 20 h after termination of nicotine infusion. They received injections of 9, 18, or 30 mg/kg L-NNA SC or vehicle alone immediately before the 20-h observation; all rats were observed for 30 min. Signs at 20 h (post-injection) as a percentage of signs at 17 h (pre-injection) declined significantly as a function of L-NNA dose. Once again, this effect was attenuated significantly more by co-administration of L-arginine than by D-arginine. The overall pattern of results suggests that nitric oxide synthesis is critical to the expression of nicotine abstinence syndrome. PMID- 9877018 TI - Resistance of neuronal nitric oxide synthase-deficient mice to cocaine-induced locomotor sensitization. AB - In brain, nitric oxide (NO) is considered as a retrograde messenger involved in synaptic plasticity. The present study was undertaken to investigate whether mice lacking the neuronal nitric oxide synthase (nNOS) gene are protected from cocaine induced behavioral sensitization. Mice were administered, IP. either saline or cocaine (15 mg/kg) for 5 days. Sensitization was determined as an increase in cocaine-induced locomotor activity on day 5 compared with day 1 and an amplified response of cocaine-experienced mice to a challenge cocaine injection given after a 10-day drug free period (e.g., on day 15). To investigate the development of a context-dependent locomotion (conditioning), the responses of cocaine- and saline experienced mice to a saline injection were determined on day 17. Male homozygote nNOS(-/-) mice were sensitive to the acute effect of cocaine (15 mg/kg) on day 1; however, they developed neither a sensitized response to cocaine (on day 5 and 15) nor a conditioned locomotion. Female homozygote nNOS(-/-) mice neither were responsive to 15 mg/kg cocaine on day 1,5 and 15, nor did they develop a conditioned locomotion. In contrast, the same cocaine regimen delivered to male and female heterozygote nNOS(+/-) mice, and wild type mice (B6 J/sv129, C57BL/6 and sv129) resulted in sensitization to cocaine-induced locomotor activity and context-dependent locomotion. Investigation of [3H]cocaine disposition in the striatum and frontal cortex of the mice revealed neither gender nor strain differences in the drug disposition. Also, no major difference in striatal dopaminergic markers between homozygote nNOS(-/-) and wild type mice was observed. The most significant distinction, however, was the finding that nNOS(-/ ) mice are completely deficient in striatal nNOS binding sites. Taken together, our results suggest that the resistance of homozygote nNOS(-/-) mice to cocaine induced behavioral sensitization is primarily due to the deletion of the nNOS gene. Considering the role of NO in synaptic plasticity, it is conceivable that reduced brain NOS activity blunts the processes that underlie the development of sensitization to cocaine. PMID- 9877019 TI - Adapting a feedforward heteroassociative network to Hodgkin-Huxley dynamics. AB - Using the original McCulloch-Pitts notion of simple on and off spike coding in lieu of rate coding, an Anderson-Kohonen artificial neural network (ANN) associative memory model was ported to a neuronal network with Hodgkin-Huxley dynamics. In the ANN, the use of 0/1 (no-spike/spike) units introduced a cross talk term that had to be compensated by introducing balanced feedforward inhibition. The resulting ANN showed good capacity and fair selectivity (rejection of unknown input vectors). Translation to the Hodgkin-Huxley model resulted in a network that was functional but not at all robust. Evaluation of the weaknesses of this network revealed that it functioned far better using spike timing, rather than spike occurrence, as the code. The algorithm requires a novel learning algorithm for feedforward inhibition that could be sought physiologically. PMID- 9877020 TI - Representation of visual space in area 7a neurons using the center of mass equation. AB - The firing rate of neurons in parietal area 7a of the behaving Rhesus monkey with its head fixed incorporates both visual and eye position information. This neural tuning is not in an ego-centered coordinate space. This physiological result was unexpected as behavioral deficits following parietal damage in human and monkey subjects suggested the existence of egocentric representations. A formulation to extract a world-centered system from area 7a neurons is presented that depends on the linearity of the eye position signal and the similarity of the equation describing the tuning of these neurons to the center of mass equation. This formulation permits the computation of the location of objects in world coordinates using either serial analysis of a single neuron's activity or parallel processing of a collection of neurons. Experimental predictions are made for the relationship between different parameters of angle of gaze neurons. PMID- 9877021 TI - A model of visuospatial working memory in prefrontal cortex: recurrent network and cellular bistability. AB - We report a computer simulation of the visuospatial delayed-response experiments of Funahashi et al. (1989), using a firing-rate model that combines intrinsic cellular bistability with the recurrent local network architecture of the neocortex. In our model, the visuospatial working memory is stored in the form of a continuum of network activity profiles that coexist with a spontaneous activity state. These neuronal firing patterns provide a population code for the cue position in a graded manner. We show that neuronal persistent activity and tuning curves of delay-period activity (memory fields) can be generated by an excitatory feedback circuit and recurrent synaptic inhibition. However, if the memory fields are constructed solely by network mechanisms, noise may induce a random drift over time in the encoded cue position, so that the working memory storage becomes unreliable. Furthermore, a "distraction" stimulus presented during the delay period produces a systematic shift in the encoded cue position. We found that the working memory performance can be rendered robust against noise and distraction stimuli if single neurons are endowed with cellular bistability (presumably due to intrinsic ion channel mechanisms) that is conditional and realized only with sustained synaptic inputs from the recurrent network. We discuss how cellular bistability at the single cell level may be detected by analysis of spike trains recorded during delay-period activity and how local modulation of intrinsic cell properties and/or synaptic transmission can alter the memory fields of individual neurons in the prefrontal cortex. PMID- 9877022 TI - Frequency control in synchronized networks of inhibitory neurons. AB - We analyze the control of frequency for a synchronized inhibitory neuronal network. The analysis is done for a reduced membrane model with a biophysically based synaptic influence. We argue that such a reduced model can quantitatively capture the frequency behavior of a larger class of neuronal models. We show that in different parameter regimes, the network frequency depends in different ways on the intrinsic and synaptic time constants. Only in one portion of the parameter space, called phasic, is the network period proportional to the synaptic decay time. These results are discussed in connection with previous work of the authors, which showed that for mildly heterogeneous networks, the synchrony breaks down, but coherence is preserved much more for systems in the phasic regime than in the other regimes. These results imply that for mildly heterogeneous networks, the existence of a coherent rhythm implies a linear dependence of the network period on synaptic decay time and a much weaker dependence on the drive to the cells. We give experimental evidence for this conclusion. PMID- 9877024 TI - Large neural network simulations on multiple hardware platforms. AB - To efficiently simulate very large networks of interconnected neurons, particular consideration has to be given to the computer architecture being used. This article presents techniques for implementing simulators for large neural networks on a number of different computer architectures. The neuronal simulation task and the computer architectures of interest are first characterized, and the potential bottlenecks are highlighted. Then we describe the experience gained from adapting an existing simulator, SWIM, to two very different architectures-vector computers and multiprocessor workstations. This work lead to the implementation of a new simulation library, SPLIT, designed to allow efficient simulation of large networks on several architectures. Different computer architectures put different demands on the organization of both data structures and computations. Strict separation of such architecture considerations from the neuronal models and other simulation aspects makes it possible to construct both portable and extendible code. PMID- 9877023 TI - Estimation of spatiotemporal neural activity using radial basis function networks. AB - We report a method using radial basis function (RBF) networks to estimate the time evolution of population activity in topologically organized neural structures from single-neuron recordings. This is an important problem in neuroscience research, as such estimates may provide insights into systems-level function of these structures. Since single-unit neural data tends to be unevenly sampled and highly variable under similar behavioral conditions, obtaining such estimates is a difficult task. In particular, a class of cells in the superior colliculus called buildup neurons can have very narrow regions of saccade vectors for which they discharge at high rates but very large surround regions over which they discharge at low, but not zero, levels. Estimating the dynamic movement fields for these cells for two spatial dimensions at closely spaced timed intervals is a difficult problem, and no general method has been described that can be applied to all buildup cells. Estimation of individual collicular cells' spatiotemporal movement fields is a prerequisite for obtaining reliable two dimensional estimates of the population activity on the collicular motor map during saccades. Therefore, we have developed several computational-geometry based algorithms that regularize the data before computing a surface estimation using RBF networks. The method is then expanded to the problem of estimating simultaneous spatiotemporal activity occurring across the superior colliculus during a single movement (the inverse problem). In principle, this methodology could be applied to any neural structure with a regular, two-dimensional organization, provided a sufficient spatial distribution of sampled neurons is available. PMID- 9877025 TI - Cytogenetics of benign breast lesions. AB - This review summarizes the cytogenetic information on benign breast lesions of various histologies, i.e., fibrocystic lesions from women with and without a known hereditary predisposition to breast cancer, fibroadenomas, phyllodes tumors, and papillomas, and relate the chromosomal features with those in breast carcinoma. In general, the frequency of chromosome abnormalities is lower in benign lesions than in breast cancer, and seems to correlate with the histologic features of the tissue, and the corresponding risk of developing invasive mammary carcinoma; aberrations are more common in proliferative than in nonproliferative lesions. The karyotypes are generally less complex than those detected in invasive carcinoma, and more often involve balanced rearrangements. No lesion specific aberration has so far been detected; on the contrary, changes repeatedly encountered in breast cancer samples can be found in benign lesions as well, e.g., gain of 1q, interstitial deletion of 3p, and trisomies 7, 18, and 20. Especially intriguing is the prevalence of rearrangements of the short arm of chromosome 3, with the minimally deleted bands 3p13-14, in proliferative lesions from prophylactic mastectomies in breast cancer families. The potential tumor suppressor gene(s) in this region remains, however, to be identified. PMID- 9877026 TI - The effect of dietary exposures on recurrence and mortality in early stage breast cancer. AB - We studied the effect of diet and body weight on recurrence and death in 472 women diagnosed with early-stage breast cancer in 1982-1984. From Cox proportional hazards regression models we found that the strongest effects were observed in premenopausal women. For example, after accounting for disease stage and age, reported baseline consumption (times/day) of butter, margarine, and lard (risk ratio (RR)=1.67; 95% confidence interval (CI)=1.17-2.39) and beer (drinks/day) (RR=1.58; 95% CI=1.15-2.17) increased the risk of recurrence. There also appeared to be an increased risk associated with consumption of red meat, liver, and bacon, corresponding to about a doubling of risk for each time per day that foods in this category were consumed (RR=1.93; 95 % CI=0.89-4.15). Relative body weight increased risk at the rate of 9% (RR=1.09; 95% CI=1.02-1.17) for each kg/m2 (equivalent to about 5.8 pounds for a woman 5'4" tall). For death, the results were similar, but relative weight was more strongly associated, increasing risk by 12% per kg/m2 (RR=1.12; 95% CI=1.03-1.22). PMID- 9877027 TI - Effects of a novel antitumor depsipeptide, FR901228, on human breast cancer cells. AB - Human breast cancer MCF7 and MDA-MB231 cells were used to investigate the biological and molecular activities of a novel naturally occurring agent, FR901228 (FR), that possesses a potent antitumor activity against human and murine tumor cells. Investigation of the cytotoxicity of FR and induction of internucleosomal DNA degradation in FR-treated cultures revealed that FR induced apoptotic-like cell death of MCF7 and MDA-MB231 cells. In FR-treated apoptotic cultures, flow cytometry revealed that there was a significant decrease of cells in S phase of the cell cycle. In FR-treated cells there was an increased expression of p21Cip1 and phosphorylation of Bcl-2 as determined by Western immunoblotting, and a novel cytoplasmic kinase of 33 kDa, p33 kinase, as determined by the in-gel kinase assay using myelin basic protein (MBP) as a substrate. Increased expression of p21CiP1, phosphorylation of Bcl-2, and activation of p33 MBP kinase may play part of the key mechanism for FR-induced apoptosis. PMID- 9877028 TI - Growth inhibition of human in vitro and mouse in vitro and in vivo mammary tumor models by retinoids in comparison with tamoxifen and the RU-486 anti-progestagen. AB - Retinoids constitute a very promising class of agents for the chemoprevention or treatment of breast cancer. These retinoids exert their biological activity through two distinct classes of retinoic acid (RA) receptors (R), the RAR isotypes (alpha, beta, and gamma) and the three RXR isotypes (alpha, beta, and gamma) and their numerous isoforms which bind as RXR/RAR heterodimers to the polymorphic cis-acting response elements of RA target genes. With respect to these numerous receptor sub-types, the retinoid-induced effects at the biological level include marked modifications with respect to both cell proliferation and cell death (apoptosis), and also in the induction of differentiation processes. The present study aims to characterize the effect which four retinoids (TTNPB, 9 cis-RA, LGD 1069, 4-HPR) with distinct RAR/RXR binding properties induced on various in vitro and in vivo mouse and human breast cancer models. The experiments with the retinoids were carried out in comparison with the anti estrogen tamoxifen and the anti-progestagen RU-486 compounds. The results show that the 6 compounds under study were markedly more efficient in terms of growth inhibition in the human T-47D cell line when maintained under anchorage independent culture conditions than when maintained under anchorage-dependent ones. While RU-486 exhibited a weak statistically significant (p < 0.05) influence on the growth of the T-47D stem cells, tamoxifen had a marked inhibitory influence on the growth of these cells. Of the four retinoids, 4-HPR was the least effective since the lowest doses tested (1 and 0.1 nM) exhibited no statistically (p > 0.05) significant influence on the growth of the stem cells. The most efficient retinoid was TTNPB. It was only at the highest dose (10 microM) that tamoxifen and RU-486 showed a weak inhibitory influence on the growth of the T-47D non-stem cells while all 4 retinoids exerted a significant inhibitory influence on the growth of these non-stem cells, with 4-HPR being the most efficient (P < 0.001) at the highest dose, but ineffective (P > 0.05) at the lowest. Tamoxifen and TTNPB were tested in vivo on hormone-sensitive (HS) and hormone-insensitive (HI) strains of the MXT murine mammary carcinoma. While TTNPB appeared to be equally efficient in terms of growth inhibition in both MXT-HS and MXT-HI models, tamoxifen had only a marginal inhibitory influence on the growth of the MXT-HI strain but did inhibit growth in the case of the MXT-HS one. TTNPB was markedly more efficient than tamoxifen in terms of both inhibiting the cell proliferation level (measured by means of computer-assisted microscopy applied to Feulgen-stained nuclei, a method which enables the percentage of cells in the S phase of the cell cycle to be determined) and triggering cell death (measured by means of the determination of the transglutaminase activity) in both the MXT-HI and MXT-HS models. The very significant TTNPB-induced inhibition of the macroscopic MXT-HS growth rate relates to the triggering of cell death (apoptosis) rather than to an inhibition of cell proliferation. All these results clearly indicate that retinoids are very efficient agents against breast cancer, at least as efficient as tamoxifen. PMID- 9877029 TI - Migration of breast epithelial cells on Laminin-5: differential role of integrins in normal and transformed cell types. AB - We examined the role of Laminin-5 (Ln-5) an extracellular matrix component of breast gland basement membrane, in supporting migration of normal (HUMEC), immortalized (MCF-10A), and malignant breast epithelial cells that exhibit different degrees of metastatic potential (MDA-MB-435>MDA-MB-231>MCF-7). HUMEC, MCF-10A, and MCF-7 cells all adhered to purified Ln-5 through the alpha3beta1 integrin receptor in adhesion assays. However, HUMEC and MCF-10A cells remained statically adherent, while MCF-7 cells migrated on Ln-5 in Transwell and colloidal gold displacement assays. Anti-alpha3 integrin antibodies blocked migration of MCF-7 cells on Ln-5. MDA-MB-231 and MDA-MB-435 cells bound and migrated on Ln-5 through a beta1 integrin receptor that is insensitive to antibodies that block the function of alpha1, alpha2, alpha3, alpha4, alpha5, alpha6, and alphaV integrin subunits. Migration of all cell types tested was blocked by CM6, a monoclonal antibody directed to a cell adhesion site on the alpha3 chain of Ln-5. Thus, Ln-5 may play an important role in regulating adhesion and migration in normal and transformed breast epithelium. Our results indicate that the type of integrin utilized by breast cells to interact with Ln 5, as well as its functional state, may determine whether cells will be statically adherent or migratory on Ln-5. PMID- 9877030 TI - Inhibition of cell proliferation by 17beta-estradiol and heregulin beta1 in estrogen receptor negative human breast carcinoma cell lines. AB - Heregulin (HRG) and 17beta-estradiol (E2) interactions that modulate growth of breast cancer cell lines have recently been demonstrated. We examined the ability of heregulin beta1 (HRGbeta1) and 17beta-estradiol to modulate the biological behavior of estrogen receptor (ER) negative human breast cancer cell lines (AU 565). The proliferation of AU-565, MBA-MB231, and SKBR3 cells was additively inhibited by treatment with 17beta-estradiol (10(-6) M) and HRGbeta1 (10 ng/ml). 17-beta estradiol did not support the transcriptional activation of a reporter gene construct containing an estrogen response element transfected into AU-565 cells. This finding suggested functional endogenous ER was not present in AU-565 cells. However, the cells contained a high number of low affinity estrogen binding sites. 17beta-estradiol only slightly decreased basal tyrosine phosphorylation of ErbB-2 and ErbB-3. Estrogen and HRGbeta1 treatment resulted in an increase of c-myc mRNA. We conclude that 17beta-estradiol and HRGbeta1, in combination, potently inhibit cell proliferation of three ER negative breast carcinoma cell lines. PMID- 9877031 TI - The cell death response to gamma-radiation in MCF-7 cells is enhanced by a neuroleptic drug, pimozide. AB - Neuroleptic drugs that bind sigma sites were tested for their ability to inhibit growth and radiosensitize MCF-7 human breast cancer cells. Inhibition of growth by approximately 50% occurred in cells exposed to pimozide (0.6 microM), haloperidol (10 microM), and the sigma ligand DTG (1,3-di(2-tolyl)guanidine, 20 microM), but no growth inhibition occurred in cells exposed to clozapine, a neuroleptic drug lacking sigma binding activity, or dextromethorphan, a selective sigma 1 binding ligand. Pimozide (2.5 microM), but not haloperidol (3.6 microM), enhanced the sensitivity of MCF-7 cells to gamma radiation in clonogenic survival assays. Pimozide significantly decreased MCF-7 clonogenic survival following a 5 or 8 Gy dose of gamma radiation, and the dose of radiation required for 1% survival (survival enhancement ratio, SER) was decreased by a factor of 2. Exposure of normal WI-38 human embryonic lung cells to pimozide did not increase their sensitivity to gamma radiation. Pimozide (2.5 microM) activated early apoptotic changes in MCF-7 cells that were detected by the uptake of Hoechst 33342 dye, and 10 microM pimozide activated a complete apoptotic pathway resulting in the death of > 90% of the cells within 24 hours. MCF-7 cells exposed to gamma radiation alone (8 Gy) showed giant cell formation, mitotic arrest, and a limited degree of apoptosis and necrosis. Within 50 hours of treatment with a combination of radiation and pimozide, cell numbers were sharply reduced compared with cultures exposed to either radiation or pimozide alone. We conclude that pimozide augmented the sensitivity of MCF-7 cells to radiation-induced cell killing through a mechanism not shared by haloperidol, but suggest that concentration of pimozide in MCF-7 cells as a result of an enrichment of sigma 2 sites might target the radiosensitization. PMID- 9877032 TI - Recommendations on the use of colony-stimulating factors in children: conclusions of a European panel. AB - During 1996 and 1997 a panel of European haematologists, oncologists, and neonatologists developed specific paediatric guidelines for the use of colony stimulating factors based on published literature and the clinical experience of these specialists within each of 13 countries. Well established indications for use comprise intervention in patients with life-threatening infection, adjunctive therapy post autologous bone marrow transplantation (BMT), mobilization of peripheral blood progenitor cells for autologous BMT, patients with acquired aplastic anaemia on anti-lymphocyte globulin and cyclosporin regimen, and severe congenital neutropenia. Less clear indications include primary prophylaxis to support dose intensification in children with high risk/advanced malignancies, secondary prophylaxis to prevent neutropenia in patients with a history of severe neutropenia, support therapy in cases of poor marrow function following BMT and for deteriorating marrow function following successful BMT, in neonatal sepsis and non infectious neonatal neutropenia, in drug induced neutropenia and in HIV positive patients. Treatment is generally well tolerated and granulocyte colony stimulating factor appears better tolerated than granulocyte and macrophage colony stimulating factor. Economically colony stimulating factors have not been shown to induce excessive costs for a given patient. CONCLUSION: In general the adult guidelines are applicable to children but additional considerations (aggressive or very progressive childhood neoplasms, specific indications, neonatal use, congenital disorders) must be taken into account. PMID- 9877033 TI - Clinical and haemodynamic correlates of heart rate variability in children with congenital heart disease. AB - Heart rate variability (HRV) represents a noninvasive parameter for studying the autonomic control of the heart. Cardiac patients have a complex autonomic disturbance. The relation of HRV to this abnormality in children with congenital heart disease (CHD) has not yet been examined. The present study examined HRV indices from 24 h Holter recordings in 258 children with an operated or non operated CHD, to determine their differences as an indicator of the severity of heart disease. The latter was defined clinically as New York Heart Association (NYHA) functional classes I to IV and haemodynamically by invasive parameters. Five time-domain measures (SDNN, SDNNi, SDANNi, rMSSD and pNN50) and three frequency-domain measures (LF, HF and balance LF/HF) were compared with normal ranges. HRV was reduced in children with CHD, except in patients of NYHA class I. The level of reduction depended on the NYHA functional class. None of the measures was significantly related to haemodynamic data. CONCLUSION: Heart rate variability is reduced in children with Congenital heart disease depending on the functional limitation but not on haemodynamic disturbances. Heart rate variability indices are sensitive markers of the clinical state. PMID- 9877034 TI - Age at onset and long-term metabolic control affect height in type-1 diabetes mellitus. AB - Reduced height as a consequence of type-I-diabetes mellitus in childhood has been reported in many studies. However, it is still debated whether good metabolic control can normalize the growth rate. A total of 436 children (204 boys, 232 girls, mean age at diagnosis of diabetes 8.2+/-0.2 years) were followed at our outpatient diabetes centre. Z-scores for height were evaluated in relation to duration of diabetes, age at onset and long-term metabolic control. At diagnosis, height in children with diabetes was significantly above the reference population (+0.43+/-0.09). Standardized height decreased during the subsequent course of diabetes. This likely represents a delay of growth, as the final height (chronological age > 18 years, n = 144) was +0.27+/-0.09. Growth reduction was more pronounced in patients diagnosed before the onset of puberty and final height in patients with a prepubertal onset of diabetes was significantly lower (+0.10+/-0.13) compared to patients with a pubertal/postpubertal onset (+0.52+/ 0.14). Among patients with a prepubertal onset, the subgroup with "poor" metabolic control (long-term median HbA(Ic) >7%) lost significantly more height compared to patients with "good" metabolic control. CONCLUSION: Despite modern treatment regimens, reduced longitudinal growth can still be demonstrated in type I diabetes. This parameter therefore provides a valuable endpoint for quality control in paediatric diabetology. PMID- 9877035 TI - Menarchal age and the stress of war: an example from Bosnia. AB - The aim of this study was to examine the influence of very low socioeconomic status, physical injury and psychological trauma on the menarchal age of deported girls who lived in besieged Srebrenica, a town that had a highly disrupted environment from the end of 1992 to mid 1995. The Srebrenica girls had a significantly higher mean menarchal age compared with a control group who lived mainly in peaceful communities in the unoccupied territory of Bosnia and Herzegovina. CONCLUSION: Our results show that psychological trauma, physical injury and low socioeconomic status, which provoked by the events of war, delay the age of menarche. PMID- 9877036 TI - Twenty-five types of T-cell receptor Vbeta family repertoire in patients with Kawasaki syndrome. AB - Recently, a possible relationship between Kawasaki syndrome (KS) and superantigen has been discussed since the report of selective expansion of specific Vbeta family in the acute phase of KS. To further investigate the relationship between KS and superantigens, we examined 25 types of T-cell receptor Vbeta family repertoire in patients with KS using the reverse transcription polymerase chain reaction. This is the first attempt to examine all of 25 Vbeta families in KS. A non radioisotope method was used to quantify mRNA so that the experiment was safer, simpler, and faster. An expression index (EI) for each Vbeta was defined as: (the amount of each Vbeta mRNA)/(the sum of all Vbeta mRNA) x 100. Ten patients with KS and ten normal children were studied. The Vbeta9 and Vbeta15 of acute phase of KS showed both significantly lower mean EI and significantly higher frequency of a decreased EI value as compared with control children. Selective expansion of the Vbeta family in the patients with KS was not observed. Although highly increased EIs were observed in various Vbetas, their frequency was not statistically significant. The pattern of increased Vbetas did not show the specific pattern that indicates a particular superantigen. CONCLUSION: In Kawasaki syndrome non-radioisotope method for analysing Vbeta mRNA is useful in cases where many samples have to be handled. The depletion of Vbeta9 and Vbeta15 or highly increased expression index in the acute phase of Kawasaki syndrome might suggest a relationship to superantigens. PMID- 9877037 TI - Antineutrophil cytoplasmic antibodies in children. AB - To define the diagnostic meaning of antineutrophil cytoplasmic antibodies (ANCA) positivity in children, we analysed 1485 consecutive sera sent for routine immunological investigation to our department from January to August 1996. Using this large screening, we identified the most typical clinical disorders associated with ANCA in childhood. Out of 1485 indirect immunofluorescence (IIF) tests for ANCA, 143 were ANCA positive, 70 had a cytoplasmic IIF pattern (c ANCA), and 73 a perinuclear IIF pattern (p-ANCA). The ANCA associated diseases in childhood were cystic fibrosis (CF) (31 c-ANCA, 7 p-ANCA positive out of 71 CF children investigated), juvenile chronic arthritis (JCA) (21 p-ANCA positive out of 78), auto-immune hepatitis (AIH) (4 c-ANCA and 12 p-ANCA positive out of 19), and ulcerative colitis (UC) (2 c-ANCA, 5 p-ANCA positive out of 15). In cases of c-ANCA positivity we determined the antigenic specificity of ANCA for proteinase 3 and/or bactericidal/permeability increasing protein. Borderline anti-proteinase 3 levels were found in CF, and in high levels in one boy with Wegener granulomatosis. Bactericidal/permeability increasing protein was characteristic target antigen in children with CF. p-ANCA positive children were further tested for the specificity for myeloperoxidase, which was detected mostly in children with JCA. CONCLUSION: The spectrum of diseases associated with ANCA in children includes, besides the diagnostic associations typical for adults, several typical pediatric entities, mainly juvenile chronic arthritis and cystic fibrosis. PMID- 9877038 TI - Acute, severe cardiomyopathy as main symptom of late-onset very long-chain acyl coenzyme A dehydrogenase deficiency. AB - A 5-year-old boy with late-onset very long-chain acyl-CoA-dehydrogenase (VLCAD) deficiency presented with acute cardiomyopathy, myopathy, gross myoglobinuria and normoglycaemia. The clinical course after diagnosis was favourable. CONCLUSION: late-onset VLCAD deficiency may present as acute cardiomyopathy. PMID- 9877039 TI - Partial N-acetylglutamate synthetase deficiency in a 13-year-old girl: diagnosis and response to treatment with N-carbamylglutamate. AB - We report on a now 13-year-old girl, who presented with recurrent episodes of vomiting, psychotic behaviour and confusion during puberty until the diagnosis of partial N-acetylglutamate synthetase deficiency was established. She had suffered one prior unclear episode of acute vomiting, lethargy and somnolence at the age of 13 months, and from childhood onward had aversion to high protein food. Treatment with a protein-restricted diet and administration of phenylbutyrate as well as L-arginine were sufficient to normalize ammonia levels but glutamine concentrations remained high. Supplementation with N-carbamylglutamate rapidly improved her protein tolerance and reduced the need for co-medication. To our knowledge, so far only seven patients with N-acetylglutamate synthetase deficiency have been reported. CONCLUSION: Partial N-carbamylglutamate deficiency has to be considered in the differential diagnosis of hyperammonaemia. If proven by enzyme determination in liver tissue, the disorder should be cautiously treated with N-carbamylglutamate. PMID- 9877040 TI - Click-evoked oto-acoustic emission measurement in preterm infants. AB - Click-evoked oto-acoustic emissions (CEOAE) are acoustic responses produced by the inner ear, reflecting functional auditory integrity. We studied both the success rate of the CEOAE method and the CEOAE presence in preterm infants during their stay at the Neonatal Intensive Care Unit (NICU), by analysis of the results of longitudinal CEOAE measurements in 19 preterm infants. The CEOAE method proved to be feasible in the majority of the population under study; the CEOAE measurement failed in 8% of the total of 226 attempts. Within the range of 30-40 weeks post-conceptional age (PCA) the CEOAE presence increased from about 50% to 80%. These results show the possible clinical application of CEOAE in a NICU for the monitoring of cochlear function during treatment of preterm infants. CONCLUSION: Bedside CEOAE measurement in preterm infants in a NICU is feasible at a PCA as low as 30 weeks. PMID- 9877041 TI - Ureaplasma urealyticum colonization and bronchopulmonary dysplasia: a comparative prospective multicentre study. AB - To determine the role of tracheal colonization at birth with Ureaplasma urealyticum and other pathogenic bacteria with regard to the development of bronchopulmonary dysplasia (BPD), 97 premature infants with very low birth weight (< 1500 g) were followed prospectively over 30 days in a multicentre study. Of those infants, 35 were colonized with Ureaplasma urealyticum (group Ia), 22 with other pathogenic bacteria (group Ib) and 40 infants with sterile tracheal aspirates served as controls (group II). Colonization with Ureaplasma urealyticum or with pathogenic bacteria independently increased the risk of developing BPD as compared to the controls (OR 2.55; 95% CI [1.11, 5.87]). Among Ureaplasma urealyticum and bacterial colonized infants, duration of mechanical ventilation and oxygen requirement were significantly longer than among controls (P < 0.05); during the interval of 11 to 35 days of life, every additional day of ventilation significantly increased the risk of BPD (OR 1.22; CI [1.12, 1.32]). The rate of oxygen supplementation, which was similar in both groups during the first 2 weeks of life, was significantly higher among the colonized infants at day 21 (0.38+/ 0.18 and 0.39+/-0.16 vs 0.31+/-0.13, P < 0.05) and at day 28 (0.38+/-0.21 and 0.34+/-0.15 vs 0.28+/-0.12, P < 0.05). For infants still ventilated at age of 28 days, Ureaplasma urealytricum and bacterial colonization were associated with a significant higher risk for BPD than for uncolonized controls (OR 5.53; [1.27, 24.02]. Association of Ureaplasma urealyticum and of bacterial colonization and BPD was not weakened after adjustments were made in a multivariate analysis for other significant risk factors. CONCLUSION: Ureaplasma urealyticum colonization is as an important risk factor in the development of bronchopulmonary dysplasia as bacterial colonization even after treatment with surfactant. PMID- 9877042 TI - Respiratory muscle rigidity in a preterm infant after use of fentanyl during Caesarean section. AB - Fentanyl is in many neonatal intensive care units the sedative of choice. One side-effect is, however, the possibility of muscle and/or chest wall rigidity. A pregnant woman with a critical pulmonary valve stenosis had a balloon dilatation at 26 weeks of gestation. She was put on propranolol, but went into a severe cardiac failure. In week 31, she developed pregnancy induced hypertension. Periodically absent diastolic flow in the umbilical cord was demonstrated. A Caesarean section was performed using fentanyl as analgesia. A boy weighing 1440 g, had a 1 min Apgar score of 3 without respiratory movements. Mask ventilation was tried, but chest wall expansion was not achieved despite using high pressures. He was intubated and positive pressure ventilation attempted, with the same result. Despite the use of high pressures up to 60-70 cm H2O, no chest movement could be achieved. An intravenous line was established in order to give naloxone and pancuronium. Just before the drugs were given, chest wall movements were achieved and the heart rate normalized. CONCLUSION: This is the first report on chest wall rigidity in a neonate after administration of fentanyl to the mother during Caesarean section. PMID- 9877043 TI - Aseptic sialadenitis in preterm infants associated with long-term oro-gastric tube feeding. AB - Sialadenitis is a rare disease in the newborn and the pathogenesis in this age group is not fully understood. We report five cases of neonatal sialadenitis in stable preterm, gavage-fed infants at 2 to 6 weeks postnatal age. The occurrence of sialadenitis was observed in temporal relation to changes in enteral feeding routines in the unit which were initiated with the objective of promoting full breast feeding at discharge. Clinical presentation consisted of a tender palpable mass over one of the salivary glands, low-grade (37.5-38.3 degrees C) fever in conjunction with clinical malaise. C-reactive protein and white blood cell count were only moderately elevated. No suppuration from salivary ducts was present. Bacterial cultures were invariably negative. Viral investigations were not carried out. Aseptic sialadenitis was suggested as the cause of the symptoms. The clinical course was benign and the local tender mass over the salivary gland vanished shortly after intravenous fluids and antibiotics were started. CONCLUSION: Long-term exclusive oro-gastric feeding may result in reduced reflex salivary gland stimulation, saliva production and hence ductal clearance of mucoid saliva, leading to functional ductal obstruction and local inflammation. PMID- 9877044 TI - Lung deposition of budesonide from turbuhaler in asthmatic children. AB - The aim of our study was to evaluate the effect of age on lung deposition of radiolabelled budesonide, delivered as a dry powder via Turbuhaler to asthmatic children. A group of 23 asthmatic children, aged 6 to 16 years, with relatively stable asthma inhaled 99mTc-labelled budesonide from a dry powder inhaler (Turbuhaler). Body and lung deposition was assessed using a gamma camera. The mean (range) median peak inspiratory flow during inhalation was 65 1 x min(-1) (45 to 76 1 x min(-1)). Mean (range) total lung deposition of 99mTc-labelled budesonide, expressed as a percentage of the metered dose, was 29.1% (15.6-47.2%) and was positively and significantly correlated with age, height and peak inspiratory flow. CONCLUSION: Total lung deposition of radiolabelled budesonide from a dry powder inhaler (Turbuhaler), is age dependent in children with moderate asthma. However, lung deposition is still satisfactory, even in younger children with lower peak inspiratory flows. PMID- 9877045 TI - Congenital tracheobronchial stenosis in monozygotic twins. AB - A pair of monozygotic twins suffering from noisy respirations and recurrent wheezes since early infancy were referred for evaluation. Using direct coronal CT and 3-dimensional reconstruction of the airway, congenital tracheobronchial stenosis was found in both twin babies, later confirmed by fibre optic bronchoscopy. The cause of congenital tracheobronchial stenosis remains obscure, but even in monozygotic twins an unidentified environmental factor cannot be excluded and may be responsible for discordant bronchopulmonary abnormalities. CONCLUSION: This case report shows that tracheobronchial stenosis may present in monozygotic twins. The pattern of malformation in twins differs from cases described previously. PMID- 9877046 TI - A newborn with cyanosis. PMID- 9877047 TI - Severe factor V deficiency presenting as subdural haematoma in the newborn. PMID- 9877048 TI - Inhaled nitric oxide does not increase lipid peroxidation in preterm infants. PMID- 9877050 TI - Validation of new blood pressure monitors for children: defects by default. PMID- 9877049 TI - Mycoplasma pneumoniae antigen detection in Guillain-Barre syndrome. PMID- 9877051 TI - Severe hypoxaemia induced by an intrapulmonary arterio-venous fistula: indication for nitric oxide. PMID- 9877052 TI - Pulmonary haemorrhage in infants with gestational age of less than 30 weeks. PMID- 9877053 TI - Sanitary status of oocytes and embryos collected from heifers experimentally exposed to Leptospira borgpetersenii serovar hardjobovis. AB - In a preliminary trial and three experiments, a total of 30 Holstein heifers were experimentally infected with a culture of Leptospira borgpetersenii serovar hardjobovis via one or more routes (uterine, cervical supraconjunctival, intranasal) and oviductal and uterine fluids recovered post-mortem or in vivo following superovulation with FSH. All routes of administration were effective in establishing Leptospira infection in the reproductive tract and Leptospira were identified in the oviductal and uterine fluids of all 30 heifers by microscopy. The incidence of infection was confirmed by positive identification of serum antibodies by the microscopic agglutination test (MAT). Twenty-one samples of the embryos (n = 59) recovered were cultured using bacteriological procedures and all tested negative for the infectious microorganism. Using polymerase chain reaction (PCR) assay, however, showed that 29% (7/24) of morula and blastocyst stage embryos, and one out of 29 oocytes tested positive for the presence of leptospiral DNA. A single oocyte or embryo collected from the infected heifers was inoculated intravenously to 26 test heifers. None of the test heifers developed antibody titers to Leptospira. It was concluded that, despite the presence of leptospires in the reproductive tract of donor animals and the association of leptospiral DNA with uterine stage embryos, the transmission of this disease is unlikely to occur by transfer of in vivo produced embryos in the bovine. PMID- 9877055 TI - Ovulation and luteal characteristics following removal of the ovine corpus luteum (lutectomy) at four times during the oestrous cycle. AB - The life span of the corpus luteum (CL) may depend on follicular development. To provide evidence relating to this hypothesis, each of 32 ewes was randomly assigned to have its CL removed on day 2, 3, 4 or 10 after oestrus. Twenty ewes were treated with 1000 IU of human chorionic gonadotrophin (hCG) 36 h after CL removal to induce ovulation; the other 12 ewes were not treated with hCG. Blood samples were collected daily to monitor the ovulatory response and the characteristics of the next cycle at the first sign of oestrus and up to day 21 after surgery or hCG administration. Every animal ovulated within 7 days of hCG administration, regardless of when its CL had been removed. It was concluded that the follicles found in the ovary as early as the second day after oestrus respond to endogenous or exogenous ovulatory stimuli affecting the life span of resulting CL. PMID- 9877054 TI - Priming effect of exogenous oestradiol on luteinizing hormone secretion in prepubertal lambs. AB - The effect of repeated administration of oestradiol-17beta on luteinizing hormone (LH) secretion was studied in 3-month old female lambs (n = 20). Animals were randomly divided in five groups and treated or not treated (group I) with 1, 2 or 3 i.m. injections of oestradiol in corn oil vehicle (1 microg/kg). Animals were slaughtered 12 (group II) and 24 h (group III) after the first injection, 24 h after the second injection (group IV) and 24 h after the third injection (group V). Animals in groups IV and V were catheterized and blood samples were collected every 4 h starting before treatment until time of sacrifice. In the rest of the groups blood samples were taken at the time of slaughter. The number of follicles > 1 mm in diameter on the ovarian surface were recorded and classified according to size. Maximum levels of oestradiol ranged from 103 to 250 pmol/l and returned to baseline within 12 to 16 h after each injection. LH secretion showed a consistent pattern in all lambs, with increases between 8 and 16 h after each oestradiol injection. The highest amplitude and longest duration (8-12 h) of LH secretion was recorded after the second oestradiol injection. Preliminary data indicated that FSH secretion resembled that of LH. There was an increase in the number of follicles with a diameter of more than 2 mm. Plasma concentrations of progesterone and cortisol were low and did not differ within groups or between treatments. The findings confirm that the pituitary LH release system in ewe lambs is sensitive to the stimulatory effects of oestradiol long before puberty. Results indicate that priming with oestradiol increases pituitary LH release to subsequent challenges of oestradiol, but long time exposure to oestradiol may have a negative effect on LH secretion. Although none of the oestradiol-treated lambs ovulated, the increase in the number of large follicles with repeated injections of oestradiol suggests that small follicles were gonadotrophin responsive and stimulated by the gonadotrophin release. PMID- 9877056 TI - Prediction of porcine semen fertility by homologous in vitro penetration (hIVP) assay. AB - A field trial was conducted to compare the fertility predicting capacity of different sperm assays applying classical semen analysis, sperm function and the homologous in vitro penetration test (hIVP) to 60 ejaculates from four boars collected over a period of 15 weeks. No differences were found between the groups of fertility (Low Fertility: < 20%; Intermediate: 40-60% and High: > 80%) for sperm-rich fraction volume collection, sperm concentration, total sperm number, cationic contents in seminal plasma and ATP concentration. Partial differences were found in the parameters of motility, normal morphology, normal apical ridge (NAR), viability with eosin-nigrosin stain, hypo-osmotic swelling test (HOS), osmotic resistance test (ORT) and functional membrane integrity (with carboxyfluorescein diacetate, DCF). These parameters would be useful for detecting sperm with poor fertility, but they are not precise enough to discriminate an ejaculate with higher fertility than the herd median. Only the penetration percentage (10.24 +/- 1.45 vs. 55.13 +/- 3.35 vs. 84.72 +/- 1.73) and sperm number per oocyte (1.29 +/- 0.07 vs. 11.29 +/- 1.79 vs. 25.86 +/- 1.43) in a hIVP system were parameters with a predictive capacity to discriminate between the three fertility groups. Consequently, hIVP was found to be the best seminal assay and it may improve the in vitro assessment of sperm fertilizing ability. PMID- 9877057 TI - Semen backflow after insemination and its effect on fertilisation results in sows. AB - The aim of the present study was to investigate the volume of and number of spermatozoa in semen backflow during and after insemination, and the effect of backflow on fertilisation results assessed at day 5 of pregnancy. Multiparous sows (n = 140) were artificially inseminated with either (1, 3 or 6) x 10(9) mixed spermatozoa from three boars in a constant volume of 80 ml. Backflow of semen was measured three times: during insemination (M1); during the first half hour after insemination (M2); and from 0.5 h until about 2.5 h after insemination (M3). Transrectal ultrasonography was performed at intervals of 4 h to determine the time of ovulation. Sows were sacrificed at 120 +/- 0.4 h after ovulation to assess the results of fertilisation. Every sow had some backflow and the variation in volume, and number of spermatozoa within the backflow was high. The average semen backflow within 2.5 h after insemination was 70 +/- 3.4% of the volume and 25 +/- 1.4% of the spermatozoa of the inseminated dosage. The concentration of the backflow (% of the inseminated dosage) decreased with time after insemination from 65% at M1 to 40% and 26% at M2 and M3, respectively. The correlations between volume and number of spermatozoa were high: r = 0.97, r = 0.73 and r = 0.81 in M1, M2 and M3, respectively. More than 5% of the inseminated spermatozoa in backflow during insemination affected fertilisation negatively in those sows inseminated with 1 x 10(9) spermatozoa (P < 0.05). Backflow after insemination had no effect on fertilisation results (P > 0.05). Timing of insemination relative to ovulation and oestrus were not related to backflow during or after insemination (P > 0.05). Of the sows which had backflow, those of parity 1 tended to have the highest proportion of sows with more than 5 ml backflow (47%; n = 8 of 17) compared with sows from parity 2 and higher (24%; n = 14 of 59) (P = 0.075). It was concluded that excessive backflow of semen during insemination had a negative effect on fertilisation results when sows where inseminated with only 1 x 10(9) spermatozoa. Causes of variation in backflow between sows were not clearly identifiable. PMID- 9877058 TI - Photoperiodic effects on pubertal maturation of spermatogenesis, pituitary responsiveness to exogenous GnRH, and expression of boar taint in crossbred boars. AB - Forty-eight weaned, winter-born crossbred males (average age of 42 days) were exposed to either a natural photoperiod (January-June at 60 degrees N, Control) or one of two artificial photoperiods (1400 lx) in light-sealed rooms. The Spring/Summer group was exposed to an artificial photoperiod simulating conditions from the vernal equinox (mid-March) to August at 60 degrees N and the Autumn/Winter group to a photoperiod, simulating conditions from the autumnal equinox (mid-September) to February at 60 degrees N. Plasma samples were collected biweekly until the pigs were slaughtered, after reaching 115 kg, and analysed for testosterone, estrone sulfate, thyroxine and prolactin. Additionally, three animals per treatment (n = 9) were injected with gonadotropin releasing hormone (GnRH) and plasma samples were collected every 15 min and analysed for luteinizing hormone and testosterone. Boar taint, carcass composition and reproductive traits were measured at slaughter. Live-weight gain from start to slaughter was lower among the Control animals compared with the Autumn/Winter and Spring/Summer animals. There was a peak in plasma testosterone in both the Spring/Summer and Autumn/Winter groups at 71 days of age, whereas plasma testosterone in the Control group remained at prepubertal levels. At 113 and 127 days of age, the Control group had somewhat higher testosterone levels than the Spring/Summer group, but at 141 days of age and on the day before slaughter, the Autumn/Winter group had a higher mean plasma testosterone concentration. There were no differences between treatments in the endocrine response to the GnRH challenge. Bulbourethral gland weight at slaughter was lower in the Spring/Summer group than in the Autumn/Winter group. The percentage of proximal cytoplasmic droplets was higher in the Spring/Summer group than in both the Control and Autumn/Winter groups. Spermatogenesis at the time of slaughter was clearly more mature in animals in the Autumn/Winter group than in those in the Spring/Summer and Control groups. Fat androstenone was lower in the Spring/Summer group than in the Control group. In the sensory evaluation, the Spring/Summer group had less boar taint than the Autumn/Winter group. Artificial short days with moderate initial changes in photoperiod, stimulated spermatogenesis compared with long days, in accordance with the pattern seen in European Wild Boars (Sus scrofa). Boar taint was also affected with higher scores in the Autumn/Winter group than in the Spring/Summer group, although this was not clearly indicated by the traditional measurements of boar taint-fat contents of androstenone and skatole. PMID- 9877059 TI - Successful embryo transfer in the silver fox (Vulpes vulpes). AB - Surgical embryo transfer in the silver fox was investigated as part of a larger project concerning the conservation of endangered canine species using modern artificial reproduction techniques with the farmed fox as a model. The animals were chosen on the basis of synchrony in natural oestrus. The timing of ovulation and artificial insemination was determined by measuring electrical resistance in the vagina. Twenty-nine embryos were flushed from eight humanely killed donor females and transferred surgically into the uteri of eight recipients. One recipient female gave birth to two male pups 47 days after the transfer of four expanded blastocysts and one embryo at the 16-cell stage derived from a donor female flushed 10 days after artificial insemination. PMID- 9877060 TI - Identification of ampullary and isthmic oviductal fluid proteins that associate with the bovine sperm membrane. AB - The objective of this study was to define the complement of ampullary and isthmic oviductal fluid proteins that associate with the sperm membrane during the non luteal and luteal stages of the oestrous cycle. Oviductal fluid was obtained from three dairy cows via indwelling cannulae in the ampulla and isthmus of the same oviduct. Daily samples of oviductal fluid were combined by region and stage of the cycle to create pools consisting of non-luteal isthmic, luteal isthmic, non luteal ampullary, and luteal ampullary oviductal fluid. An aliquot from each oviductal fluid pool was biotinylated by incubation in biotin reagent. Spermatozoa from three fertile bulls were pooled, washed and incubated for 4 h in each of the four pools of biotinylated oviductal fluid. After incubation, sperm membrane proteins were solubilized, and proteins subjected to one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. The biotinylated oviductal fluid proteins that associated with the sperm membranes were detected by avidin-horseradish peroxidase and diaminobenzidine substrate. Eight oviductal fluid proteins with apparent molecular weights of 97, 75, 66, 55, 48, 34, 28 and 24 kDa were consistently detected to associate with the sperm membrane. Differences in ampullar and isthmic fluid proteins that associated with sperm during the luteal and non-luteal stage of the oestrous cycle were not detected. PMID- 9877061 TI - Treatment of gilts with leukocytes from the sire does not improve reproductive performance. AB - The effectiveness of pretreatment of gilts with leukocyte antigens on reproductive performance was studied. Two experiments were carried out that involved the treatment of gilts with leukocytes prior to artificial or natural insemination. Gilts were randomly assigned to one of three treatment groups. Group A gilts received two doses of 2 x 10(8) sire's leukocytes; one injected intra-peritoneal at first estrus and the other infused into the uterus immediately prior to insemination at the second estrus cycle. Group B gilts were similarly treated with the 'gilt's own' (autologous) leukocytes and Group C animals received phosphate buffered saline as controls. The same boars were used as sires across all three treatment groups. Gilts were slaughtered during the fifth week of pregnancy and their reproductive performance assessed. When data from the three groups were compared there were significant differences in embryo survival and placental weights between the sire's leukocyte treatment and PBS control groups. Embryo survival rate (No viable embryos/No corpora lutea x 100) at the fifth week of gestation was lower (p < 0.05) and placental weights were higher in the sire's leukocyte treatment group (p < 0.05). However, the latter may likely be due to fewer developing embryos. Embryo survival and placental weights did not vary significantly between the sire's and autologous leukocytes treatment groups. The treatment of gilts with leukocytes from the boars used as sires showed no improvement on subsequent reproductive outcome over that observed for the autologous leukocytes or PBS control treatments. Consequently, in a healthy well managed herd leukocytes treatment offers no advantage to pig reproductive function. PMID- 9877062 TI - Seminal plasma does not advance ovulation in hCG-treated sows. AB - In gilts, seminal plasma treatment before or during the LH-surge has been found to advance ovulation in all animals by as much as 8 to 14 h. Two experiments were performed to assess whether such an advancement occurs in multiparous sows in which ovulation is induced by 750 i.u. hCG at 68 h after weaning. In both experiments, seminal plasma was inseminated at 4, 5 and 6 h after hCG (7 and 6 sows, respectively) and control sows (6 and 6 sows, respectively) were not inseminated. In Experiment 1, using Meishan semen, all sows ovulated between 38 and 44 h after hCG; no advancement of ovulation was seen due to treatment. In Experiment 2, using GY seminal plasma, 3 and 4 sows, respectively had started ovulation at 44 h after hCG. Again, no advancement of ovulation was seen due to treatment. Therefore, in both experiments, seminal plasma treatment within 4-6 h after hCG failed to advance ovulation to a similar extent as found in spontaneously ovulating gilts. It is unclear what causes this lack of effect. Maybe seminal plasma treatment does not advance hCG-induced ovulation or batches of seminal plasma differ in their ovulation-advancing properties. PMID- 9877063 TI - Concentrations of gonadotropins, estradiol and progesterone in sows selected on an index of ovulation rate and embryo survival. AB - The objective of this study was to determine concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P4) and 17beta estradiol (E2) in sows from a line selected on an index which emphasized ovulation rate (Select) and from a control line. A further classification of the sows in each line was made according to the estimated number of ovulations during an estrous cycle. Sows in the Select line were ranked into a high (HI) or low group (LI) when their estimated number of ovulations were 25 or more and 14 to 15, respectively. Sows of the control line were classified into groups as high (HC) or low (LC) when the estimated values for ovulation rate were 14-15 and 8-9 ovulations, respectively. Blood samples were collected every 12 h during a complete estrous cycle and samples were analyzed for concentrations of FSH and LH. Samples collected every 24 h were assayed for P4 and E2. Mean concentrations of FSH, LH, P4 and E2 did not differ (P > 0.10) between lines or between HI and LI or HC and LC groups. Selection of pigs for ovulation rate and embryonal survival did not affect concentrations of FSH, LH, P4 and E2 in sows during the estrous cycle. PMID- 9877064 TI - The effect of pH on the motility of spermatozoa from chicken, turkey and quail. AB - At 40 degrees C in a NaCl-based buffer the motility of spermatoza from chicken, turkey and quail was inhibited at pH values below 7.8, 7.2 and 7.2, respectively. At these pH values the percentage motile and velocity of spermatoza were relatively low, but the motility became vigorous when the pH was raised by 0.2 units and increased even more following further alkalinization. Spermatozoa from all three species stored motionless at pH 6.0 for 3 h could be reactivated by dilution in an alkaline solution (pH 9.0). These findings support the hypothesis that a change in the environmental pH could be implicated in the suppression and stimulation of sperm motility during oviducal sperm storage and transport. PMID- 9877066 TI - Experimental transmission of Hepatozoon americanum Vincent-Johnson et al., 1997 to dogs by the Gulf Coast tick, Amblyomma maculatum Koch. AB - Experimental transmission of Hepatozoon americanum to dogs was attempted with four ixodid ticks, viz., Rhipicephalus sanguineus, Amblyomma americanum, Amblyomma maculatum and Dermacentor variabilis. Ticks that dogs ingested included some that were laboratory-reared and experimentally fed as nymphs on a dog with naturally occurring hepatozoonosis; other ticks were collected as replete or partially engorged larvae, nymphs and adults from dogs that had hepatozoonosis and natural infestations of ticks. Whole ticks used to expose susceptible dogs orally were partially dissected to help release oocysts. Among eight dogs exposed, only the three that were fed A. maculatum adults experimentally acquisition fed as nymphs became infected. Dogs developed elevated body temperature and other evidence of clinical disease starting 4 weeks after exposure. 'Cysts' typical of H. americanum were found in skeletal muscle when samples were first examined 5 weeks after dogs ingested ticks, and parasites were also observed in peripheral blood smears at approximately the same time. Our study demonstrates that A. maculatum nymphs can acquire H. americanum by feeding on a parasitemic dog and that transstadial transmission of the protozoan occurs, with dogs acquiring infection when they ingest newly molted adult ticks. PMID- 9877065 TI - Localisation of low molecular weight sperm antigens during capacitation and acrosome reaction. AB - FITC-labelled sperm-specific antibodies against hamster spermatozoa were utilized as probes in acrosome reaction assays. An indirect immunofluorescence test demonstrated the localisation of two sperm proteins of 19 kDa and 23 kDa on the anterior acrosomal cap region of washed cauda epididymal sperm. These proteins were not detected in reacted acrosome or on immature or immotile sperm. Antisperm agglutinating antibodies specific to these two low molecular weight sperm antigens could be useful probes for evaluating the acrosomal status of mammalian spermatozoa. PMID- 9877067 TI - Production, faecal egg counts and worm burdens of ewe lambs which grazed six contrasting forages. AB - Lambs were grazed on monospecific swards of one of six forages, some containing condensed tannins (CT); sulla (Hedysarum coronarium), Lotus corniculatus and L. pedunculatus and some without CT (lucerne (Medicago sativa), plantain (Plantago lanceolata) and a ryegrass/white clover pasture (Lolium perenne/Trifolium repens)) to evaluate effects of forage type on gastrointestinal nematode burdens, faecal egg count as well as liveweight gain (LWG), wool growth and dag formation over a 42-day period. A new allotment of feed was provided at seven-day intervals, and feed was available ad libitum. Total worm burdens in a pre treatment slaughter group were ca. 22000 of which ca. 5000 were in the abomasum. Within each forage type, 10 of the 25 lambs (NP) were given an anthelmintic drench at 14-day intervals and the remaining 15 lambs (P) were not drenched. Daily gains of the NP lambs were double that of P lambs which grazed either L. corniculatus, lucerne or pasture, but parasitism had a lesser effect on performance of lambs which grazed sulla and L. pedunculatus. Plantain was not palatable and all lambs performed poorly. Highest daily gains in P lambs were for those which grazed sulla (175 g/day) and L. pedunculatus (160 g/day) with total worm burdens of 13100 and 23000 for the respective treatment groups. The other forages resulted in lower daily LWG in P lambs, and performance was not related to either worm burdens or worm species. This experiment has shown that, when sulla is fed, there is a reduction in worm burdens and faecal egg count (FEC), but with Maku lotus which also contains CT, the good level of performance was achieved despite in high worm burdens and FEC. The mechanisms by which these forages enable high levels of productivity in the face of a parasitism appear to differ, but both could be incorporated into forage feeding systems to reduce dependence on anthelmintic drenches. PMID- 9877069 TI - A hemolytic factor from Haemonchus contortus alters erythrocyte morphology. AB - A hemolytic factor from adult Haemonchus contortus caused distinct morphological changes in the surface of sheep red blood cells (RBCs). After a 15 min exposure to the hemolytic factor, hemolysis was not detected in incubation media, but RBCs were spherical in shape with numerous surface projections compared to control cells that were smooth-surfaced biconcave disks. After 30 min, a time at which significant hemolysis occurred, echinocytes were formed, and after 90 min, cells were severely disrupted with many visible holes in membranes. No RBC ghosts were observed. RBCs from four other mammalian species were lysed by the H. contortus hemolytic factor. However, the rate of hemolysis varied with a relative order of sheep approximately rabbit>goat>pig>calf. The morphology of RBCs from all four species was significantly altered after 30 min incubation with the degree of morphological changes related to the degree of hemolysis. These results support the hypothesis that the hemolytic factor acts as a pore-forming agent, although a phospholipase or other enzyme might play a role in solubilization of cell membranes. PMID- 9877068 TI - Comparative efficacy evaluation of moxidectin gel and ivermectin paste against internal parasites of equines in Brazil. AB - A total of 24 male and female equines of mixed breed, 10-20 months of age and naturally infected with internal parasites was utilized in a controlled test to evaluate the efficacy of a moxidectin 2% gel formulation at the dosage of 0.4 mg moxidectin per kg of live weight and an ivermectin 1.87% commercial paste formulation at the dosage 0.2 mg ivermectin per kg applied orally. Animals were allocated into three groups of eight horses each based on pre-treatment eggs per gram (EPG) counts and treatments were randomized among the groups. One group was kept as untreated controls. One animal in the moxidectin-treated group died before the end of the trial from a cause unrelated to treatment leaving a total of seven animals in this group. Fecal egg counts were performed three times post treatment and the number of parasites remaining in each animal was determined. Statistical analyses using geometric means were performed at the 1% level of significance. Both moxidectin and ivermectin preparations reduced initial EPG from a mean of 1600 to 0 on Days 5, 7 and at the end of the trial on Day 14. Efficacy percentages of moxidectin and ivermectin against immature and adult nematodes were as follows: Trichostrongylus axei, Parascaris equorum, Strongylus edentatus, S. vulgaris, Triodontophorus spp. and Gyalocephalus capitatus, 100% for both products; Habronema muscae 99.5 and 99.6%, respectively, Strongyloides westeri, 100 and 99.2%, respectively; Oxyuris equi, 99.6 and 100%, respectively; small strongyles, 99.7% for both products. Of the latter, the most numerous were: Cylicocyclus insigne, Cylicostephanus longibursatus and Cyathostomum catinatum. No Gasterophilus nasalis were found in horses from either treated group, while two of eight control horses had infections with thisparasite. Moxidectin showed greater efficacy (84.9%) than ivermectin (67.8%) against Strongylus vulgaris larvae found in the mesenteric artery aneurisms, but the difference was not statistically significant. Total parasite counts for both treated groups were significantly lower (p<0.01) than in the non-treated group. No significant differences were noted between moxidectin and ivermectin. Efficacy against the 30 nematode species found in this study was very evident for both products. As expected, neither moxidectin nor ivermectin was effective in controlling the tapeworm Anoplocephala perfoliata. No adverse reactions were observed during the experimental period. PMID- 9877070 TI - Natural evolution of gastrointestinal nematodes in goats (Capra hircus) in the semi-arid ecosystem of the Paraiba backwoods, northeastern Brazil. AB - This study was conducted in Patos county, a semi-arid area with caatinga vegetation in Paraiba State, northeastern Brazil. Twenty-four male goats of mixed breeds were used. The animals were separated in age groups from 1-12 months, i. e., 1-2, 3-4, 5-6, 7-8, 9-10, 11-12 months, with four animals in each group. The animals were slaughtered and necropsied to recover and identify adult worms. The goats were exposed to nematode infection from the first month of life. The most prevalent nematodes in decreasing order were: Trichostrongylus colubriformis, Haemonchus contortus and Oesophagostomum columbianum. Age of the goats did not appear to be a factor in resistance to gastrointestinal nematode infection. Worm burdens of 11-12 month old animals were significantly (P<0.05) higher than in other age groups. PMID- 9877072 TI - Localization and migration route of Cephalopina titillator (Diptera: Oestridae) larvae in the head of infested camels (Camelus dromedarius). AB - The localization and migration routes of Cephalopina titillator larvae were studied in the heads of 45 naturally infested camels. Larvae were located in the pharyngeal cavity (95.6%), the labyrinth of the ethmoid bone (71.1%), the turbinates (28.9%) and the lower nasal meatus (6.7%). Out of 261 first-instar larvae, 73.7% were found in the labyrinth of the ethmoid bone and 26.3% in the turbinates, while 88.9% of second-instar larvae were observed in the pharyngeal cavity and 11.1% in the labyrinth of the ethmoid bone. Most third-instar larvae (91.9%) were found in the pharyngeal cavity, with a few larvae in the labyrinth of the ethmoid bone (6.5%) and lower nasal meatus (1.6%). The first moult occurred in the labyrinth of the ethmoid bone. The second moult was observed in both the labyrinth of ethmoid bone and the pharyngeal cavity. The migration route of these larvae in the infested head was discussed. PMID- 9877073 TI - Seroprevalence of Toxoplasma gondii in Egyptian donkeys using ELISA. AB - A serological survey of Toxoplasma gondii was conducted using ELISA on 121 sera from adult donkeys from Monofia Province, Egypt. Antibodies to T. gondii were found in 65.6% of 121 donkeys. PMID- 9877071 TI - Characterization of protective immune responses in local lymphoid tissues after drug-attenuated infections with Ostertagia ostertagi in calves. AB - In the present study, cell-surface markers and cytokine gene expression of lymphocytes from the local lymph nodes were studied 9 days after primary infection with Ostertagia ostertagi in previously naive calves or in calves previously immunized with multiple, chemically attenuated infections. Changes in lymphocyte populations were assessed by flow cytometry utilizing monoclonal antibodies specific for bovine cell-surface markers. Changes observed in the percentages of lymphoid populations after challenge were similar in animals immunized by either three or five drug-attenuated infections. In both immunized groups, the CD4+/CD8+ ratio was significantly higher than in naive animals after challenge infections. In addition, both immunized groups showed significantly lower levels of Ig-bearing cells upon experimental challenge when compared to animals with a primary experimental infection. No differences were observed in the number of gammadelta or interleukin 2 receptor (IL-2R) positive cells. The levels of mRNA for IL-4, IL-10, IL-15, IFN-gamma and TGF-beta1 were examined by competitive RT-PCR. After challenge, the levels of these cytokines were lower in animals immunized by five drug-attenuated infections, and in the case of IL-4 and TGF-beta1, these differences were statistically significant. These results indicate that animals exhibiting protection from reinfection with O. ostertagi do not show a shift to higher percentages of Ig+ cells characteristic of a primary infection. In addition, protected animals appear to show a decreased IL4 and TGF beta1 response upon challenge when compared to non-immune animals. PMID- 9877074 TI - Habitat related differences in helminth parasites of red foxes in the Ebro valley. AB - The prevalences of helminth parasites found on red foxes collected in two different habitats of the Ebro Valley, in north-eastern Spain were compared. Twenty helminth species, including one trematode, six cestodes, 12 nematodes and one acanthocephalan were found. Metorchis albidus, Uncinaria stenocephala and Dirofilaria immitis were more prevalent in the irrigated lands close to the main rivers of the study area, whereas Trichuris vulpis and Pterygodermatites affinis showed higher prevalences in the semiarid stepparian habitat. Habitat characteristics need to be considered when surveying helminths over large geographical areas. The study shows that in the Ebro Valley the fox constitutes a reservoir host for several helminth species that are parasites of the domestic dog or the wolf, or may have importance as zoonoses. PMID- 9877076 TI - Ca2+ channel antagonists and neuroprotection from cerebral ischemia. AB - Stroke is the third leading cause of death and the main disabling neurologic disease. The finding in experimental studies that neuronal death does not occur immediately after ischemic injury has encouraged the development of neuroprotective agents. Various Ca2+ channel antagonists, that is, L-type selective or non-selective derivatives from classical Ca2+ channel antagonists, have been examined for their ability of neuroprotection through improvement of cerebral blood circulation or inhibition of Ca2+ overload induced by excessive glutamate release. Although some of the antagonists showed efficient neuroprotection in animal models, systemic hypotension limited the utility of these drugs, and none of the compounds showed beneficial effects in treatments for acute ischemic stroke in clinical trials. Drugs other than Ca2+ channel antagonists developed on the basis of the glutamate-Ca2+ overload hypothesis were shown also to lack clinical benefit. Recently, some mechanisms have been proposed to interpret neuronal death in relation to hyperexcitability or apoptosis after ischemic insult. In these hypotheses, activation of the Ca2+ channel types selectively expressed in neuronal tissues is proposed as a critical step of the pathways toward neurodegeneration. Thus, it is increasingly recognized that developing highly selective compounds for neuronal Ca2+ channels is not only important for treatment of stroke but also for elucidation of mechanisms that underlie neurodegeneration. PMID- 9877075 TI - Severe cat flea infestation of dairy calves in Brazil. AB - An investigation was conducted into a severe flea infestation on dairy calves. Inspection of the dairy revealed a high infestation of Ctenocephalides felis felis on four calves and thousands of fleas in the stable where cows and calves were brought in for milking. A 3-month-old female calf was highly infested with fleas. The animal showed an evident lethargy, weight loss, as well as pale mucous membranes and dehydration. Hematological analysis revealed anemia, with 10% packed cell volume and a 1.72 x 10(6) microl(-1) erythrocyte count. Blood transfusions were performed and the flea infestation was controlled with 1% fipronil pour-on (1 ml/10 kg). The farmer was advised to treat the other calves with fipronil, remove the manure from the stable and spread 1% propoxur powder (100 g/m2) onto the floor of the stable. PMID- 9877077 TI - Effect of subchronic metrifonate treatment on cerebral glucose metabolism in young and aged rats. AB - The effects of subchronic administration of metrifonate, a long-lasting cholinesterase inhibitor, on local cerebral glucose utilization were assessed in 3- and 27-month old Sprague-Dawley rats, using the autoradiographic [14C]2 deoxyglucose technique. Rats were treated twice daily with metrifonate (80 or 120 mg/kg) for 3 weeks. The [14C]2-deoxyglucose experiment was performed 18 h after the last metrifonate administration. In 3-month old rats, metrifonate 80 mg/kg increased the average hemispheric cerebral glucose utilization by 12% (P > 0.001). Significant effects were observed in 19 of the 54 regions studied, including cortical and limbic regions. The higher dose induced a larger effect (average increase 17%, 24 of the 54 regions affected). In 27-month old rats, very similar effects were obtained. These results show that repeated administration of metrifonate leads to a sustained metabolic activation in rat brain, at a level comparable to the activation observed previously after a single administration of the drug. PMID- 9877078 TI - The dopamine D2 receptor antagonist sulpiride causes long-lasting serotonin release. AB - The effects of the dopamine D2 receptor antagonist sulpiride on extracellular levels of serotonin (5-hydroxytryptamine, 5-HT) and the 5-HT metabolite 5 hydroxyindoleacetic acid (5-HIAA) were examined by using in vivo voltammetry. Sulpiride (1 or 3 mM, 2 microl over 24 min) was administered to freely moving rats via a cannula implanted in the striatum and 5-hydroxyindole levels were measured by using a carbon fiber voltammetry electrode implanted in the ipsilateral striatum. Six to 8 h after injection, 5-hydroxyindole levels increased 3-fold, peaked 1 to 2 days post-injection, and returned to normal levels within 2 to 4 days. These effects were suppressed by pretreatment with p chlorophenylalanine. Two days after sulpiride injection, high-performance liquid chromatography of striatal homogenates revealed that although the 5-HT concentration was unchanged, the 5-HIAA concentration was increased significantly. These results suggest that the long-lasting elevation of 5 hydroxyindole concentrations was primarily due to increased 5-HT release. PMID- 9877079 TI - Sympathetic vasoconstriction in the rat anterior choroid is mediated by alpha1 adrenoceptors. AB - These experiments were undertaken in an attempt to use laser-Doppler flowmetry to measure anterior choroidal blood flow in the anesthetized rat and to study the mechanism by which sympathetic nerve stimulation might produce vasoconstriction in this vascular bed. Electrical stimulation of the preganglionic cervical sympathetic nerve produced reproducible, frequency-related ocular vasoconstrictor responses with maximal vasoconstriction seen at about 32 Hz. Ocular vasoconstrictor responses were blocked by intravenous treatment with the nonselective alpha-adrenoceptor antagonists, phentolamine (5 mg kg(-1)) and phenoxybenzamine (2 mg kg(-1)), as well as with the selective alpha1-adrenoceptor antagonist, prazosin (0.3 mg kg(-1)). In contrast, the selective alpha2 adrenoceptor blocker, rauwolscine (0.5 mg kg(-1)), only potentiated the vasoconstriction. Neither intravenous atropine (1 mg kg(-1)) nor propranolol (1 mg kg(-1)) altered the magnitude of neurally evoked vasoconstriction. These results demonstrate the usefulness of laser-Doppler flowmetry in studies of the rat anterior choroidal circulation and suggest that adrenergic neurogenic vasoconstriction in the anterior segment of the rat eye is mediated almost exclusively by alpha1-adrenoceptor mechanisms. PMID- 9877080 TI - The choleretic effects of N-acetylglucosaminides, major urinary metabolites of ursodeoxycholic acid, in bile fistula rats. AB - We investigated the effects of three bile acids conjugated with N acetylglucosamine, ursodeoxycholate N-acetylglucosaminide, tauroursodeoxycholate N-acetylglucosaminide and glycoursodeoxycholate N-acetylglucosaminide, on bile flow and biliary excretion of various markers in comparison with ursodeoxycholic acid, tauroursodeoxycholic acid and glycoursodeoxycholic acid in bile fistula rats. These bile acids were infused intravenously at a constant rate of 0.3 or 0.6 micromol/min/100 g b.w. for 2 h. All bile acids examined increased bile flow in a dose-dependent manner. In particular, ursodeoxycholate N-acetylglucosaminide has a longer-lasting effect after its infusion on bile flow than the other bile acids. Furthermore, these bile acids markedly increased biliary total bile acid excretion. At a higher dose level, the coefficient of determination (r2) between the biliary total bile acid excretion and bile flow for ursodeoxycholate N acetylglucosaminide (r2 = 0.39) was lower than that for the other bile acids (r2 = 0.75-0.92). The ursodeoxycholate N-acetylglucosaminide, as well as tauroursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholate N acetylglucosaminide and glycoursodeoxycholate N-acetylglucosaminide, was mostly excreted in an unchanged form in bile, whereas ursodeoxycholic acid was excreted as a conjugate with taurine. The three N-acetylglucosaminides as well as ursodeoxycholic acid, tauroursodeoxycholic acid and glycoursodeoxycholic acid significantly increased the biliary excretion of cholesterol, phospholipid, bilirubin and total Ca2+. In contrast, the N-acetylglucosaminides significantly decreased in biliary bicarbonate concentration, whereas ursodeoxycholic acid significantly increased biliary bicarbonate concentration. However, tauroursodeoxycholic acid and glycoursodeoxycholic acid did not significantly change the biliary bicarbonate concentration. The results indicate that N acetylglucosaminides have a choleretic effect in bile fistula rats. Our present study also demonstrates that N-acetylglucosaminides, but not ursodeoxycholic acid, tauroursodeoxycholic acid or glycoursodeoxycholic acid, can significantly reduce the biliary bicarbonate concentration. Furthermore, our findings suggest that ursodeoxycholate N-acetylglucosaminide may partly exert a choleretic effect via mechanisms different from those of the other bile acids. PMID- 9877081 TI - Antiemetic effects of sendide, a peptide tachykinin NK1 receptor antagonist, in the ferret. AB - The antiemetic activity of sendide, a new peptide tachykinin NK1 receptor antagonist, against cisplatin-induced emesis was investigated using ferrets. The frequency of cisplatin (10 mg/kg, i.p.)-induced retching (104.6 +/- 14.3/6 h) and vomiting (19.0 +/- 3.0/6 h) was significantly reduced by pretreatment with sendide (3.0 mg/kg, s.c.) (14.0 +/- 8.1/6 h and 1.8 +/- 1.2/6 h, respectively). Intravenous bolus injection of substance P (1-10 microg/kg) or 5 hydroxytryptamine (5-HT) (10-50 microg/kg) produced a dose-dependent increase in the abdominal afferent vagus nerve activity. The change from pre-injection level in the afferent nerve activity induced by substance P (1 microg/kg, i.v.) (453.7 +/- 51.5%) was significantly reduced by pretreatment with either sendide (100 microg/kg, i.v.) (276.1 +/- 50.1%, P < 0.05) or granisetron, a 5-HT3 receptor antagonist (1 mg/kg, i.v.) (146.3 +/- 14.0%, P < 0.01). The amount of 5-HT released into the solution during a 1-h exposure to 2-methyl-5-HT (10(-6) M), a 5 HT3 receptor agonist, was significantly increased (317.9 +/- 46.7%, P < 0.05) compared with that of the control tissues (160.4 +/- 8.1%). The 2-methyl-5-HT induced 5-HT release was significantly inhibited by administration of sendide (10(-6) M) (174.0 +/- 21.6%, P < 0.05) or granisetron (10(-6) M) (186.6 +/- 27.3%, P < 0.05). Since sendide does not penetrate the central nervous system, these results suggest that the antiemetic effects of sendide are due to the inhibition of NK1 and 5-HT3 receptors on the emetic peripheral detector sites. PMID- 9877082 TI - Protective effects of melatonin in zymosan-activated plasma-induced paw inflammation. AB - The aim of the present study was to investigate the protective effect of the pineal hormone melatonin in a model of acute local inflammation (zymosan activated plasma-induced paw oedema), in which oxyradicals, nitric oxide (NO) and peroxynitrite are known to play a crucial role in the inflammatory process. The intraplantar injection of zymosan-activated plasma elicited an inflammatory response that was characterized by a time-dependent increase in paw oedema, neutrophil infiltration and increased levels of nitrite/nitrate in the paw exudate. The maximal increase in paw volume was observed at 3 h after administration (maximal in paw volume: 1.34 +/- 0.09 ml). At this time point, myeloperoxidase activity and lipid peroxidation were markedly increased in the zymosan-activated plasma-treated paw (226 +/- 10.2 mU/100 mg wet tissue, 31 +/- 2.1 mM/mg wet tissue, respectively). However, zymosan-activated plasma-induced paw oedema was significantly reduced in a dose-dependent manner by treatment with melatonin (given at 62.5 and 125 microg/paw) at 1, 2, 3, 4 h after injection of zymosan-activated plasma. Melatonin treatment also caused a significant reduction of the myeloperoxidase activity and lipid peroxidation and inhibited nitrite/nitrate levels in the paw exudate. The paw tissues were also examined immunohistochemically for the presence of nitrotyrosine (a marker of peroxynitrite formation). At 3 h following injection of zymosan-activated plasma, staining for nitrotyrosine was also found to be localised in the inflamed paw tissue. Treatment with melatonin (125 microg/paw) reduced the appearance of nitrotyrosine in the tissues. Our findings support the view that melatonin exerts anti-inflammatory effects. PMID- 9877083 TI - Alpha2B-adrenoceptors couple to Ca2+ increase in both endogenous and recombinant expression systems. AB - The ability of cloned human alpha2B-adrenoceptors heterologously expressed in Sf9 cells and endogenous alpha2B-adrenoceptors in NG 108-15 neuroblastoma x glioma cells to couple to increase of intracellular Ca2+ was studied. Ca2+ increases in NG 108-15 cells were detectable but slight, whereas those in alpha2B-adrenoceptor expressing Sf9 cells were greater. In the latter, the maximum Ca2+ increase correlated positively, and the EC50-value of noradrenaline negatively, with the receptor expression density. The order of potency of the agonists was D medetomidine ([D]-4-[5]-[1-(2,3-dimethylphenyl)ethyl]-1H-imidazole) > noradrenaline approximately = clonidine > oxymetazoline, with clonidine and UK14,304 (5-bromo-N-[4,5-dihydro-1H-imidazole-2-yl]-6-quinoxalinamine) being weak partial agonists. In Sf9 cells Ca2+ increases consisted of concomitant mobilization from an intracellular store and influx of extracellular Ca2+. In these cells alpha2B-adrenoceptor stimulation also increased the inositol 1,4,5 trisphosphate mass. We conclude that alpha2B-adrenoceptors can couple to intracellular Ca2+ increases which may involve prior activation of phospholipase C. PMID- 9877084 TI - Lobeline inhibits Ca2+ current in cultured neurones from rat sympathetic ganglia. AB - The effect of lobeline was studied on the voltage-activated Ca2+ current in sympathetic neurones from the rat superior cervical ganglia using the whole-cell variant of the patch-clamp technique. Lobeline (10-300 microM) inhibited the Ca2+ current evoked by voltage steps from -80 mV (holding potential) to 0 mV (test potential) in a dose dependent manner. The inhibitory effects of noradrenaline (10 microM) and lobeline (100 microM) were compared using a prepulse protocol with high (+80 mV) depolarization. Within the same cell depolarizing prepulses decreased the inhibitory effect of noradrenaline but did not change the extent of lobeline inhibition. Addition of GTPgammaS (300 microM) to the pipette solution did not prevent the inhibitory effect of lobeline (100 microM) but greatly reduced that of noradrenaline (100 microM). Our experiments suggest, that the weak nicotinic agonist lobeline exerts a direct blocking effect on Ca2+ channels at concentrations commonly used to release transmitters. PMID- 9877086 TI - Expert systems for the interpretation of serum proteins. AB - The ready availability of inexpensive desk-top computers with enormous disk storage has made the practicality of computer assisted medical interpretive software a reality. There seems little question that these programs could be of enormous help to physicians. However, there are daunting problems to their creation, including the lack of standards for clinical diagnostic precision or accuracy and paucity of helpful literature. As a result, the final products may be quite different. Little effort has been devoted in the laboratory to produce programs which could have great benefit in bridging the gap between laboratorians and clinicians. In a few circumscribed areas where the interpretation of laboratory measurements have been well studied in relation to patient demographics and to the final outcome, the impact has been enormous. The prime example is prenatal diagnosis of neural tube defects, and certain chromosomal and developmental abnormalities. Viewed as an obstacle by most people able and willing to attempt to create such programs is the omnipresence of necessary regulation. A brief overview of the general structure of a program to assist with the interpretation of serum proteins is given as a model in the perspective of current knowledge and state of the relevant literature. PMID- 9877085 TI - Inhibition by local anesthetics of Ca2+ channels in rat anterior pituitary cells. AB - The characteristics of local anesthetic inhibition of voltage-dependent Ca2+ channels in a rat pituitary clonal cell line were investigated by whole-cell voltage clamp and compared with inhibition by the dihydropyridine Ca2+ channel antagonist, nicardipine. With extracellular Ba2+ (10 mM) as the current carrier, depolarization above -40 mV evoked a slowly inactivating I(Ba). Extracellularly applied lidocaine inhibited I(Ba) without changing the activation threshold, the voltage of peak current, or the reversal potential. Inhibition was greater at a holding potential of -60 mV (IC50 = 1.2 mM) than at -80 mV (IC50 = 2.6 mM). This depolarization-induced potentiation in I(Ba) inhibition developed over 0.1-10 s after membrane depolarization began. Nicardipine also dose-dependently inhibited I(Ba) with an IC50 = 90 nM (at a holding potential = -80 mV). Both lidocaine and nicardipine shifted the I(Ba) steady-state inactivation (availability) curves to the left. Double-pulse protocols revealed that lidocaine (1 mM) accelerated the depolarization-induced inhibition (inactivation) of I(Ba) over the rate in drug free solutions, but had no effect on the hyperpolarization-induced removal of channel inactivation. Nicardipine also accelerated the depolarization-induced inactivation of I(Ba) but, in addition, it slowed the hyperpolarization-induced inactivation removal. The relative inhibitory action of lidocaine in suppressing I(Ba) was unchanged in the presence of nicardipine. These results suggest that lidocaine has a direct action on membrane Ca2+ channels, similar to the voltage dependent action of dihydropyridine, but acting at a separate and independent site. PMID- 9877087 TI - Proteome in perspective. AB - This review describes briefly proteome science. It explains why proteome science or proteomics emerged only recently and why a shift from genomics to proteomics is occurring. This review further illustrates that proteomics can unravel new domains in nature's complexity. Finally, it demonstrates that proteomics is offering new tools for the study of complex biological or medical problems. PMID- 9877088 TI - Usefulness of chromogranin A as a marker for detection of relapses of carcinoid tumours. AB - It is well known that peptide-producing endocrine tumours cosecrete immunoreactive chromogranin A with their characteristic hormones. Into this study 187 patients with the diagnosis malignant carcinoids or other malignancies were entered. Using chromogranin A at a cut-off level of 30.3 U/ml it was possible to discriminate between patients in remission and patients suffering a relapse with a sensitivity of 91.7% and a specificity of 96.4%, which may be of important diagnostic value. In our study that lasted over one year we could clearly show that the measurement of chromogranin A is impressively superior to 5 hydroxyindoleacetic-acid for detecting a relapse of carcinoids. PMID- 9877089 TI - Comparison of tissue polypeptide antigen (TPA) with cancer antigen 15-3 (CA 15-3) and carcinoembryonic antigen (CEA) in follow-up of breast cancer. AB - Cancer antigen 15-3 (CA 15-3), carcinoembryonic antigen (CEA) and tissue polypeptide antigen (TPA) were measured in 679 sera of breast cancer patients and in 94 sera of women without breast cancer. The tumour markers were determined using immunoluminometric assays (ILMA). The assays are characterised by an inter assay-imprecision and intra-assay-imprecision <4 %. The breast cancer patients were staged according to the TNM classification stage 0-IV (by UICC) in patient groups with a compatible prognosis. Median and range of each stage were investigated. The cut-off values (95th and 97.5th percentile of control group) of CA 15-3, CEA and TPA were determined; specificity, sensitivity, positive and negative predictive value (PV) and efficiency were investigated for these cut off's and the receiver operating characteristic (ROC) curves were calculated. The differences between control group and stage 0-3 were shown as non-significant for CA 15-3 and CEA but significant for TPA. Significant differences were found in stage 4 for all three tumour markers. The three tumour markers did not have differences in specificity, positive and negative PV and efficiency. TPA and CA 15-3 demonstrated comparable results in sensitivity and ROC curve analyses. These results were better than those from CEA. PMID- 9877090 TI - Continuous analysis in extracorporeally circulating blood--a rat model applying flow-through ion-selective electrodes for the measurement of Ca2+, K+, Na+ and pH. AB - A rat model is introduced which enables investigations in anticoagulated blood with continuous measurements by a flow-through electrode system. In the present study, a potentiometric ion-selective electrode (ISE)-system was used for measuring Ca2+, K+, Na+ and pH in rats. The setup was adjusted to an extracorporeal blood-volume of 0.750 ml. This permits indirect measurements of the analytes via a dialysis membrane, with electrical separation of the ISE's and the animal. The flow-rates of blood and dialysis-solution were adjusted in such a way that water diffusing from the aqueous dialysis solution into the blood, across the dialysis membrane, does not alter the haematocrit. Polyethyleneglycol hirudin was used for anticoagulation, since it was superior to heparin. The assembly enables continuous measurements in the living anaesthetized rat over a time period of at least 3 hours. PMID- 9877091 TI - Improved specificity of a new direct assay for urinary cortisol: application in corticoid treated patients. AB - Twenty four hours urinary free cortisol (UFC) excretion has been determined in 35 eucortisolic control patients, in seven of them before and after tetracosactide (Synacthen) stimulation and in 18 patients treated by anti-inflammatory steroids. Results of the new direct Immunotech RIA (DIm) were compared to those of the INCSTAR RIA kit with (ECA) or without methylene chloride extraction (DCA). In controls DIm UFC (106.2+/-45.8 nmol/24h) was significantly lower than DCA UFC (397+/-119 nmol/24h) and than ECA UFC (127+/-49 nmol/24h). After tetracosactide stimulation, median of DCA/DIm ratio decreased from 3.61 to 1.88 whereas ECA/DIm ratio did not change significantly (1.31 to 1.06). In treated patients most DCA and ECA results were over the upper limit of controls but only 5 DIm results were increased. DIm assay showed good specificity and practicability and may be used with benefit in the evaluation of the adrenal gland function. PMID- 9877092 TI - Quantitative automated particle-enhanced immunonephelometric assay for the routinary measurement of human cystatin C. AB - Human cystatin C is a low molecular mass protein of 13359 Dalton recently proposed as a new very sensitive marker of changes in glomerular filtration rate. Serum cystatin C concentration correlates negatively with glomerular filtration rate as well as or better than creatinine. We evaluated a recently introduced automated nephelometric immunoassay for cystatin C in serum or EDTA-plasma samples on the Behring Nephelometer System. The assay consists of incubating the 100-fold diluted sample for 6 minutes with latex particles covalently coated with anti-human cystatin C antibodies, and then quantifying the change of light scatter produced. Method reproducibility is satisfactory, the intra- and inter assay coefficients of variation ranging from 1.58% to 3.77% and from 5.6% to 11.47% respectively. Rheumatoid factor (< or = 1116 IU/ml), bilirubin (< or = 418 micrommol/l), triglycerides (10.47 mmol/), and haemoglobin (12 g/l) do not significantly interfere in the assay. No significant difference was found in cystatin C concentration between serum and EDTA-plasma samples. Cystatin C is stable in serum samples stored under different conditions up to one month. This method correlates well (mean difference=-0.536+/-0.307 mg/l) with another commercially available particle-enhanced turbidimetric immunoassay. Cystatin C offers better clinical sensitivity than creatinine for discriminating patients with normal renal function and those with mild-to-moderate reduction in renal function. This method is suitable for routine cystatin C measurement, including emergencies. PMID- 9877093 TI - External Quality Assessment of stat test intralaboratory turnaround times. Pilot study from the Members of the Working Group for the Standardization and Promotion of Turnaround Time Control under the Auspices of the Comitato Italiano per la Standardizzazione dei Metodi Ematologici e di Laboratorio. AB - We describe procedures, results and prospects of a pilot program in External Quality Assessment (EQA) of the stat test intralaboratory turnaround times. Our goals are to promote quality by systematic monitoring and comparison of performances by laboratories, continuous investigation into the state of the art of the processes from receipt of sample to transmission of results and creation of a data base for standardization of measures and definition of consensus values for turnaround time. Of 30 laboratories invited to participate, 25 took part, agreeing to record times of arrival and transmission for all determinations of three analytes (blood hemoglobin, serum/plasma potassium and plasma prothrombin time) for seven consecutive days and to continue for one or more further periods of seven days as necessary if there were less than 300 determinations for each analyte. Within a preset time limit, data were sent by e-mail on an Excel file and we sent back two reports per analyte, showing: i) the graph for time vs. percentage of tests completed and several measures of turnaround time; ii) results of all laboratories in graph form, allowing each laboratory to identify only its own data. The high proportion of participating laboratories among those invited (83%) encourages us to implement the EQA program systematically, on a half-yearly basis, extending it to all laboratories wishing to participate in Italy or elsewhere in Europe. PMID- 9877094 TI - Biological effects of eleven combined oral contraceptives on serum triglycerides, gamma-glutamyltransferase, alkaline phosphatase, bilirubin and other biochemical variables. AB - The objectives of this paper are to update and quantify the biological effects of the most commonly used oral contraceptives (OC) on 15 biochemical tests currently determined in clinical laboratories and to compare these effects between the different types of OC. The sample population was constituted by 1604 women using combined OCs and the control group comprised 3466 women in the same age range not taking medication. Women taking OC were divided into 11 groups according to the estrogen/progestogen combination. The effects of OCs were studied after adjustment for age, weight, height, body mass index and alcohol and tobacco consumption. The changes observed with the new progestogens were less important than in the past. In comparison with the controls, the mean serum triglyceride concentration was significantly increased by +8.5% to +36.0% (p<0.05 to p<0.001) in each group while those of total cholesterol and gamma-glutamyltransferase were increased only in 3 and 4 estrogen/progestogen combinations respectively. Conversely, the mean concentrations of alkaline phosphatase, total bilirubin, phosphate and albumin were significantly decreased. Using a discriminant analysis, three main groups according to the type of progestogen were defined: cyproterone acetate, DL-norgestrel and levonorgestrel, and all other progestogens. The changes in serum triglyceride concentration induced by OC intake must be considered by the clinician and are useful for taking a clinical and risk decision in an individual woman. PMID- 9877095 TI - Diagnostic and prognostic value of biochemical markers in malignant bone disease: a prospective study on the effect of bisphosphonate on pain intensity and progression of malignant bone disease. AB - Seventy cancer patients with malignant osteolytic bone disease received pamidronate every three weeks for a maximum of six cycles. Bone resorption parameters, urinary calcium excretion, and pain parameters were assessed at baseline and throughout the study. At baseline, 80-95% of patients showed elevated urinary pyridinoline, deoxypyridinoline, Osteomark NTx and serum ICTP levels, whereas only 35% of patients had elevated urinary CrossLaps excretion rates. During bisphosphonate therapy, significant decreases in Osteomark NTx, CrossLaps and calcium excretion were observed, which were not related to the clinical outcome. The baseline levels of bone resorption markers were used to predict the probability of non-progressive bone disease or reduction in pain intensity during bisphosphonate therapy. Significant predictors of non progressive bone disease were urinary pyridinoline and serum ICTP levels; significant predictors of reduction in pain intensity were urinary free deoxypyridinoline and serum ICTP levels. Our data indicate that serum ICTP levels predict significantly the response to bisphosphonate therapy in patients with advanced malignant osteolytic bone disease. CrossLaps did not predict the clinical outcome, but decreased significantly during bisphosphonate therapy. Our data demonstrate that the different bone resorption markers are reflecting different aspects of bone metabolism, and therefore differ in their diagnostic and prognostic properties. PMID- 9877096 TI - IFCC Committee on Standardization of Markers of Cardiac Damage: premises and project presentation. International Federation of Clinical Chemistry and Laboratory Medicine. AB - The field of biochemical markers of cardiac damage is in a dynamic state, with new applications continually appearing and new assays and markers being developed. These significant and sudden advancements in the development of new biochemical assays have led however to several analytical and interpretative problems. In this situation, it is essential that a uniform and rigorous outlook be maintained to ensure optimal test utilization. For these reasons, the IFCC Scientific Division recently agreed to establish a Committee on "Standardization of Markers of Cardiac Damage" (C-SMCD), inviting members from the established American and European groups to become members of this Committee. In this presentation, the premises, the issues, and the proposed plan of action of C-SMCD are presented and discussed. PMID- 9877097 TI - Does alpha-MSH have a role in regulating skin pigmentation in humans? AB - Over the years there has been much debate as to whether alpha-MSH has a role as a pigmentary hormone in humans. There are two main reasons for this. First, despite the observations in the 1960s that alpha-MSH increased skin darkening in humans, there are reports that the peptide has no effect on melanogenesis in cultured human melanocytes. Second, the human pituitary, unlike that of most mammals, secretes very little alpha-MSH and circulatory levels of the peptide in humans are extremely low. However, there is now evidence from several groups that alpha MSH is capable of stimulating melanogenesis in cultured human melanocytes. Rather than producing an overall increase in melanin production, it appears that the peptide acts specifically to increase the synthesis of eumelanin. Such an action could well explain the previously observed skin darkening effects of alpha-MSH. It is also now known that alpha-MSH is not produced exclusively in the pituitary but has been found at numerous sites, including the skin where it is produced by several cell types. Related Proopiomelanocortin (POMC) peptides such as ACTH are also produced in human skin. The ACTH peptides act at the same receptor (MC-1) as alpha-MSH and certain of these would appear to be more potent than alpha-MSH in stimulating melanogenesis. The ACTH peptides are also present in greater amounts than alpha-MSH in human epidermis and it is likely that they play an important role in regulating pigmentary responses. These POMC peptides are released from keratinocytes in response to ultraviolet radiation (UVR) and it has been proposed that they serve as paracrine factors in mediating UV induced pigmentation. Their production by keratinocytes could therefore be critical in determining pigmentary responses and any changes in the availability of these POMC peptides might explain the variations in tanning ability seen in different individuals. However, the possibility that tanning ability is also dependent upon differences at the level of the MC-1 receptor cannot be ruled out and it has been suggested that an inability to tan may depend upon the presence of non-functional changes at the MC 1 receptor. alpha-MSH does, of course, affect human melanocytes in several ways and its stimulation of melanogenesis could be the consequence of some other fundamental action in the melanocyte. The peptide also has many other target sites in the skin and while it may have a role in regulating skin pigmentation in humans, it should not be viewed solely as a pigmentary peptide. alpha-MSH clearly has many different actions and its primary role in the skin may be to maintain homeostasis. PMID- 9877098 TI - Enhanced melanogenesis induced by tyrosinase gene-transfer increases boron-uptake and killing effect of boron neutron capture therapy for amelanotic melanoma. AB - Specific and powerful cancer killing effect for melanoma by boron neutron capture therapy (BNCT) using DOPA analogue, 10B-p-boronophenylalanine (10B-BPA), has been established, but amelanotic melanoma is insufficiently responsive to 10B-BPA BNCT in comparison with actively melanin-producing melanoma. Although the accumulation mechanism of 10B-BPA within melanoma was not established, we have recently obtained findings suggesting that melanin monomers, key intermediates for melanin polymer formation, play a critical role in 10B-BPA accumulation. In addition, there are some kinds of human amelanotic melanomas, such as MEL2A, in which expression of tyrosinase is repressed or lacking though tyrosinase-related protein (TRP)-1 and TRP-2 are well expressed. Thus, by using a similarly tyrosinase-lacking mouse amelanotic melanoma cell line, A1059, we constructed TA1059 cells by transfecting human tyrosinase-cDNA into these cells. TA1059 cells acquired higher DOPA-oxidase and DOPAchrome tautomerase activity as well as eumelanin content at even higher levels than those of B16F10 cells. TA1059 cells showed about 2.5 times higher P-boronophenylalanine (BPA) uptake than A1059 cells in culture. In animal experiments, by using these cell lines, tumor growth of TA1059 was significantly suppressed by 10B-BPA BNCT as compared with A1059. These findings indicate that the induction of active melanin biosynthesis by melanogenic gene-transfer effectively improves the treatment of amelanotic melanoma by BNCT. PMID- 9877099 TI - The tyrosinase gene from medakafish: transgenic expression rescues albino mutation. AB - We have determined the 9.8 kb genomic nucleotide sequence of the tyrosinase gene and its 5 upstream region from a teleost, medakafish (Oryzias latipes), and shown that the coding region is composed of five exons and four introns, spanning 4.7 kb. While the number and sizes of the exons were found to be similar to those of mammalian tyrosinase genes, however, the total size of the coding region (4.7 kb) was demonstrated to be less than one tenth those of mouse (ca. 70 kb) and human (> 70 kb) genes. Primer extension analysis revealed that the transcription initiation site starts with a long untranslated leader sequence (340 nucleotide long) from the AUG start codon. A characteristic CATGTG sequence known as a putative regulatory motif in melanocyte-specific genes was present in the 131st base upstream from the initiation site, while other typical regulatory elements such as the TATA-box or M-box common to terrestrial vertebrates were lacking. Transgenic experiments were carried out by microinjecting two kinds of plasmid clones into fertilized eggs of the albino i(l) mutant: one consisting of the genomic tyrosinase gene with the 10 kb 5 upstream region and the other the tyrosinase cDNA with the 3 kb 5 upstream region. The results showed that 53 and 45 of 114 and 118 transgenic eggs, respectively, developed normally beyond hatching and 15 and 10 exhibited a mosaic pattern of pigmentation. Despite the absence of typical regulatory elements like a TATA-box in both cases correct melanin pigmentation was obtained without ectopic expression. Thus, transgenic expression rescued from the albino-i(l) mutation, and the i locus of the medaka genome can be concluded to encode the tyrosinase gene. PMID- 9877100 TI - Development of melanocyte progenitors in murine Steel mutant neural crest explants cultured with stem cell factor, endothelin-3, or TPA. AB - Stem cell factor (SCF) has been suggested to be indispensable for the development of neural crest cells into melanocytes because Steel mutant mice (i.e., Sl/Sl(d)) have no pigmented hairs. On the other hand, it has been demonstrated that the addition of endothelin 3 (ET-3) or TPA to neural crest cell cultures can induce melanocyte differentiation without addition of extrinsic SCF. In this study, we excluded the influence of intrinsic SCF by using Sl/Sl mouse embryos to study more precisely the effects of natural cytokines, such as extrinsic soluble SCF or ET-3, or chemical reagents, such as TPA or cholera toxin. We found that SCF is supplied within the wild-type neural crest explants and that ET-3 cannot induce melanocyte differentiation or proliferation without SCF. These results indicate that SCF plays a critical role in survival or G1/S entry of melanocyte progenitors and that SCF initially stimulates their proliferation and then ET-3 accelerates their proliferation and differentiation. TPA has the ability to elicit neural crest cell differentiation into melanocytes without exogenously added SCF but it is not as effective as SCF because many more melanocytes developed in the wild-type neural crest explants cultured with TPA. PMID- 9877101 TI - Melanoma x macrophage fusion hybrids acquire increased melanogenesis and metastatic potential: altered N-glycosylation as an underlying mechanism. AB - We recently reported that a majority of hybrids generated in vitro between weakly metastatic mouse Cloudman S91 melanoma cells and human or mouse macrophages showed enhanced metastatic potential. With few exceptions, hybrids with enhanced metastatic potential also had elevated basal melanin content and increased responsiveness to MSH compared to parental cells. Here we investigated the hybrid melanotic phenotype in more detail, comparing the pigmentary systems of hybrids and parental Cloudman S91 cells by several techniques. Cells were studied by electron microscopy, cell lysates were analyzed for tyrosinase (E.C.1.14.18.1) activity, and melanosomal proteins were analyzed by gel electrophoresis and immunoblotting. Melanosomes in parental Cloudman melanoma cells were few in number and relatively amorphous, whereas those in the hybrids were numerous and heavily pigmented, containing highly organized lattice structures. Both basal and MSH-inducible tyrosinase activities were elevated several fold in hybrids compared to parental cells. Tyrosinase, TRP-2, and LAMP-1 from hybrids migrated more slowly on gels compared to the same proteins from parental melanoma cells, consistent with increased glycosylation. Migration of LAMP-1 from hybrids was similar to that from peritoneal macrophages, which also appeared to be more heavily glycosylated than LAMP-1 from Cloudman cells. By using 3H-glucosamine as a marker of N-glycosylation, its incorporation into tyrosinase and LAMP-1 was found to be elevated in hybrids, suppressed by N-glycosylation inhibitors, and stimulated by MSH to a greater degree in hybrids compared to parental cells. These results indicate N-glycosylation as an important regulatory pathway for MSH induced melanogenesis and further suggest that altered N-linked glycosylation may be an underlying mechanism for regulation of both melanogenesis and metastasis in macrophage x melanoma hybrids. PMID- 9877102 TI - Agouti-related maturation and tissue distribution of alpha-Melanocyte Stimulating Hormone in wild-type (AwJ/AwJ) and mutant (Ay/a,a/a) mice. AB - Because of ectopic overproduction of agouti protein, yellow alleles (A(y) and A(vy)) of the murine agouti gene may secondarily modulate the synthesis, maturation (i.e., acetylation), and/or tissue deployment of alpha-Melanocyte Stimulating Hormone (MSH). We used HPLC to test the hypothesis that A(y)/a mice exhibit altered concentrations of desacetyl-, monoacetyl-, and diacetyl-alpha-MSH in pituitaries, sera, and telogen hair bulbs when compared to black (a/a) mice. We also used RIA to measure total MSH in those same tissues of A(y)a,a/a, and white-bellied agouti (A(wJ)/A(wJ)) mice (Strain C57BL/6J). We found no evidence that A(y)/a mice possessed an imbalance of des-, mono-, and diacetylated alpha MSH species. However, radioimmunoassay (RIA) analyses of total MSH suggest that wild-type agouti mice (A(wJ)/A(wJ)) exhibited significantly decreased (P < 0.05) tissue levels of total alpha-MSH in pituitaries, sera, and regenerating hair bulbs when compared to those of mutant A(y)/a and a/a mice. PMID- 9877103 TI - Cytogenetic follow-up in a case with a primary cutaneous melanoma and five metastatic lesions. AB - Cytogenetic analyses conducted on several cases of melanoma have contributed to the identification of the chromosomal regions where the sequences responsible for malignant transformation and the evolution of this tumor are probably located. With regard to these problems, it is very important to have the possibility to analyze, through the use of cytogenetics, both the primary melanoma and the metastatic lesions from the same patient. We present a case in which the primary melanoma and five different metastases were studied by using cytogenetics. The primary tumor showed an inversion of chromosome 1 where the p36 region, often proposed in literature as the location of a melanoma susceptibility gene, was involved. Three cutaneous and one lymphonodal metastases presented the same nine clonal chromosomal aberrations. In particular, one is a further rearrangement of the marker present in the primary tumor; another is a deletion of the 9p21pter region in which the p16 gene is located. Our results can provide a contribution to the hypothesis of the location of a candidate gene for melanoma in the 1p36 region and can also underscore the role of the 9p21 region in the progression of melanoma. PMID- 9877104 TI - Changes in lipid peroxide level in retinal pigment epithelial cells in vitro upon addition of linoleic acids or linoleic acid hydroperoxides under varying concentrations of oxygen. AB - We previously observed the presence of autofluorescent lipofuscin or its like in retinal pigment epithelial (RPE) cells, which were incubated with linoleic acid hydroperoxides (LHP). We studied the effect of oxygen on the level of lipid peroxides in RPE cells in the presence of linoleic acids (LA) or LHP. The level of lipid peroxides in these cells was determined by use of the thiobarbituric acid-reactive substance (TBARS), which responded to oxygen concentrations qualitatively, and a linear regression analysis. Multiple linear regression analysis disclosed that treatment with LA for 24 hr resulted in detectable increase in the level of TBARS in the cells, whereas treatment with LA or LHP for 48 hr caused detectable decrease. Stepwise linear regression analysis showed that the level of TBARS decreased in an oxygen-tension dependent manner in the cells incubated with LA for 48 hr. Thus, it was shown that short-term incubation with LA increased the level of TBARS in the cells and that LA decreased its level in an oxygen-tension dependent manner. For these results, the postulation was made that the prolonged auto-oxidation of LA caused production of lipofuscin-like materials, a complex of lipid peroxides and proteins that were insoluble in SDS and acetic acid solution. PMID- 9877105 TI - Plasma thrombomodulin in renal disease: effects of renal function and proteinuria. AB - Plasma thrombomodulin (PTM) may be a marker of vascular endothelial damage and is increased in active vasculitis. However, since PTM is a mixture of glycoproteins (MW between 28 to 105 kD), some may be variably excreted by the kidney. PTM level thus may be affected both renal impairment and proteinuria. This study examines patients with varied renal pathology and relates PTM levels to renal function and proteinuria. PTM levels were measured in eighty nine renal patients with varied renal pathology: 23 patients on hemodialysis (HD), 66 with variable renal function (from normal to severe renal impairment), and proteinuria from insignificant to nephrotic range. PTM levels rose as renal function declined and were highest in HD patients. PTM levels also seemed to rise with increasing proteinuria. Indeed, this relationship appeared to be exaggerated in patients with hypertensive and diabetic nephropathy compared to those with primary glomerulonephritis. Measurements of PTM are clearly affected both by renal function and proteinuria. Thus the confidence limits above which an abnormal level is recognised should be increased as renal function declines. PMID- 9877106 TI - Glomerular filtration rate measurement in cirrhotic patients with renal failure. AB - Assessment of renal function, namely glomerular filtration rate (GFR), by the renal creatinine clearance, may be problematic and less accurate in cirrhotics; however, it is an essential element in the global evaluation of these patients. In order to better characterize misinterpretation of GFR by the renal creatinine clearance, we compared a group of cirrhotic patients with renal failure (n - 30) to a group of chronic renal failure patients (CRF) without liver disease (n - 5). Inulin and PAH clearances were measured during a 4-hour infusion of inulin and PAH; renal creatinine clearance was measured during a 24-hour urine collection and this, simultaneously with inulin and PAH clearance. We observed that in moderate to severe renal failure (renal inulin clearance 30 +/- 10 ml/min), GFR (i. e. renal inulin clearance) in cirrhotic patients was overestimated by the renal creatinine clearance, similarly to CRF patients (ratio 1.8 +/- 0.7 and 1.6 +/- 0.9 respectively); however cirrhotic patients have a lower serum creatinine (186 +/- 97 vs 133 +/- 62 micromol/l respectively). On the other hand, cirrhotic patients with mild renal dysfunction (renal inulin clearance 74 +/- 15 ml/min) had a renal creatinine clearance of 77 +/- 25 ml/min. Systemic inulin clearance overestimated renal inulin clearance, proportionally to the severity of renal dysfunction. We conclude that it is only the degree of renal failure, irrespective of etiology, that explains the overestimation of the glomerular filtration rate by the renal creatinine clearance in cirrhotic patients; hence, there is no specific alteration in the renal excretion of creatinine in cirrhotic patients. PMID- 9877107 TI - Patients with analgetic nephropathy on chronic hemodialysis have a high incidence of severe secondary hyperparathyroidism. AB - Between 1996 and 1997, 86 patients were treated for terminal renal failure by hemodialysis at Maribor Teaching Hospital. Among them were 12 with iPTH over 900 pg/ml and symptoms of bone disease. In these patients bone biopsy was carried out with the aim of determining the type of renal osteodystrophy (RO) and establishing a possible correlation with the clinical picture, with densitometry and laboratory results. Histomorphologically, 6 patients fulfilled the criteria for secondary hyperparathyroidism (HT) - 3 with analgetic nephropathy (AN), one with chronic pyelonephritis (CPN), one with vascular nephropathy (VN), one with chronic glomerulonephritis (CGN). Six patients fulfilled the criteria for mixed osteodystrophy (MO) - 3 AN, 2 CGN, one VN. According to laboratory findings and bone mineral density (BMD), a statistically significant difference between HT and MO was present only in AP (Table 1). The most frequent diagnosis in patients with iPTH >900 pg/ml was analgetic nephropathy. PMID- 9877108 TI - Acute hyponatremia in the perioperative period: insights into its pathophysiology and recommendations for management. AB - Our purpose is to review the topic of acute postoperative hyponatremia by focusing on pertinent aspects of the physiology of water and solute excretion. Four areas will be highlighted: an examination of the source of addition of electrolyte-free water, an exploration of the basis for the very large natriuresis that occurs during cerebral salt wasting following neurosurgery, possible reasons to explain why acute postoperative hyponatremia may pose a greater risk for young women [Ayus and Arieff 1996, Ayus et al. 1992, Arieff 1986, Wijdick et al. 1991], and issues related to treatment of acute hyponatremia. PMID- 9877109 TI - The effect on blood pressure of an acute fall in ionized calcium during hemodialysis. A randomized study in two patients. AB - AIM: Elevating serum ionized calcium levels is known to stabilize blood pressure during and after a hemodialysis session. The data on lowering calcium levels is limited. SUBJECTS AND METHODS: This study examined the responses to an acute drop in ionized calcium during a four hour hemodialysis session in two subjects who differed in the presence (patient 1) or absence (patient 2) of anti-hypertensive medication and average weight loss of 4 kg (patient 1) or 2 kgs (patient 2) with each dialysis. Parathyroid hormone (PTH) levels were normal (patient 1) or very high (patient 2). A calcium free dialysate was used and the calcium infusion rate distal to the dialyzer was varied to produce either no change in ionized calcium for 10 dialyses, or a fall of at least 0.2 mmol/l for 10 dialyses. The sequence was randomized and was blinded to the observers and patients. RESULTS: The fall in ionized calcium was similar in each individual, 0.37 +/- 0.11 and 0.34 +/- 0.05 mmol/l. PTH rose from 8.6 +/- 1.6 to 24.2 +/- 6.6 pmol/l for patient 1 and 144.6 +/- 59.9 to 234.8 +/- 32.3 pmol/l for patient 2: patient 1 showed a fall in blood pressure after dialysis associated with the fall in calcium. This was most pronounced in the upright position whereas there was no change for Patient 2. For Patient 1 standing mean blood pressure post-dialysis was 104 +/- 6 mmHg when the calcium was stable and 94 +/- 10 mmHg with hypocalcemia p <0.01. Mean blood pressure during dialysis was also lower with hypocalcemia 100 +/- 7 versus 92 +/- 9 p <0.05. There were no significant changes in blood pressure for patient 2. Neither of the patients had any symptoms attributable to hypotension or hypocalcemia. CONCLUSION: It was concluded that modest falls in ionized calcium were associated with a drop in standing blood pressure only when combined with other factors predisposing to vascular instability. Even so these decreases were small enough to be asymptomatic. PMID- 9877110 TI - Maintenance hemodialysis in patients with HIV-associated nephropathy. AB - OBJECTIVE: End-stage renal disease (ESRD) patients infected with human immunodeficiency virus (HIV) have poor survival on maintenance hemodialysis. Only a few studies have evaluated survival time on the basis of demographic and clinical factors. The clinical category of the HIV infection and total CD4 counts are commonly considered determining factors of survival in these HIV-infected dialysis patients. PATIENTS AND METHODS: A retrospective case review of all ESRD patients with HIV infection on maintenance hemodialysis, from January 1987 through December 1996, was performed to determine the impact of different clinical categories of HIV infection and CD4 counts on survival and to see if there are other factors that can predict survival among these patients. From a total of 75 ESRD patients with HIV infection, 58 patients with ESRD due to HIV associated nephropathy (HIVAN) on maintenance hemodialysis are reported here. RESULTS: During the 10 year study period, 52 of 58 ESRD patients with HIVAN expired. Infection (60%), cardiogenic conditions (13%), cerebro-vascular accidents (6%), HIV wasting (8%) and noncompliance with dialysis (11%) were common causes of death. Fifty patients who were on long term hemodialysis (Group I), had a median survival time of 11 months (4-69). Among 44 diseased patients in Group I, various demographic, clinical and laboratory markers, including age, sex, race, acquired immunodeficiency syndrome (AIDS)-associated conditions, HIV clinical categories, hemodialysis access and initial serum albumin level were not significantly associated with mean or median survival time. Those with initial CD4 counts of more than 50 had a significantly longer median survival (11.3 months) than those whose counts were below 50 (5.3 months). Patients with < or = 2.5 g/100 ml initial serum albumin level and < or = 50 initial CD4 counts had a median survival time of 5.3 months compared to 13.6 months in the group of patients with initial serum albumin level of > 2.5 g/100 ml and initial CD4 counts > 50. Both of these findings were statistically significant. CONCLUSIONS: Our 10 year experience of maintenance hemodialysis in ESRD patients with HIVAN shows that long term survival is possible. Initial CD4+ T cells of < or = 50 in these patients is a poor prognostic marker. HIV clinical categories, as reported by others, failed to predict survival in our long term experience. Initial serum albumin of < or = 2.5 g/100 ml was associated with poor survival, though statistically not significant. When initial serum albumin of < or = 2.5 g/100 ml was combined with CD4+ T cells of < or = 50, it became another marker of poor survival. PMID- 9877111 TI - The effects of increasing exchange volume and frequency on peritoneal dialysis adequacy. AB - AIM: Peritoneal dialysis adequacy guidelines are generally not met with the standard prescription of continuous ambulatory peritoneal dialysis (CAPD), four 2 liter (1) exchanges daily. The aim of this study is to determine the effects of increasing exchange volume singularly or in combination with frequency on peritoneal dialysis adequacy. PATIENTS AND METHODS: Fourteen stable ESRD patients receiving peritoneal dialysis were evaluated for adequacy and nutritional status between the fourth and sixth months during a six-month baseline period in which the dialysis prescription was four two-liter exchanges daily and during a six month intervention period in which patients (n = 7) from group 1 were prescribed CAPD four 2.5-liter exchanges daily while patients (n = 7) from group 2 were prescribed continuous cycling peritoneal dialysis (CCPD - 12 l) using four 2.5 liters during the night and a 2-liter wet day. RESULTS: Mean total weekly urea Kt/V (TWKt/V) increased significantly from 1.6 +/- 0.2 to 2.1 +/- 0.2, p <0.01 in group 1, and from 1.6 +/- 0.4 to 2.1 +/- 0.5, p <0.001 in group 2. Mean normal total weekly creatinine clearance increased significantly from 51 +/- 11 to 60 +/ 8 l/1.73 m2, p <0.05 in group 1, and from 45 +/- 6 to 58 +/- 9 l/1.73 m2, p <0.01 in group 2. Serum albumin of almost all patients in the intervention groups were higher than in the baseline groups. Mean serum albumin increased from 3.6 +/ 0.4 to 4.0 +/- 0.4 g/dl, p <0.01 in group 1, and from 3.8 +/- 0.2 to 4.0 +/- 0.4, p <0.05 in group 2. The magnitude of the decrement in BUN and serum creatinine were greater in group 2 than group 1 (p <0.001 and p <0.05, respectively). When the two intervention groups were compared to each other, no significant differences in the delivered dialysis dose or nutritional status were noted. CONCLUSION: In conclusion, it is possible to achieve currently proposed adequacy target by increasing the exchange volume singularly or in combination with frequency in most peritoneal dialysis patients. PMID- 9877112 TI - Simultaneous lowering of serum phosphate and LDL-cholesterol by sevelamer hydrochloride (RenaGel) in dialysis patients. AB - The aim of the current investigation was to study the effects of sevelamer hydrochloride (RenaGel) on serum phosphate, intact parathyroid hormone levels (iPTH), and lipid profiles in stable hemodialysis patients. Hemodialysis patients maintained on calcium containing phosphate binders were enrolled in this study. Following two weeks of washout of the phosphate binders, serum phosphate rose from 6.4 +/- 0.6 to 10.5 +/- 0.7 mg/dl (p <0.001). After 8 weeks of titration with sevelamer hydrochloride, serum phosphate fell by 4.5 +/- 0.3 to 6.3 +/- 0.7 mg/dl (p <0.0001). Serum calcium levels fell during washout (9.8 +/- 0.4 to 8.9 +/- 0.3 mg/dl, p <0.004) and were unaffected by sevelamer hydrochloride. Sevelamer hydrochloride administration was associated with a 23.0 +/- 3.1% fall in total cholesterol, a 35.9 +/- 3.0% fall in LDL cholesterol, and a 35.2 +/- 5.3% fall in the LDL:HDL cholesterol ratio (p <0.001). There was no change in HDL cholesterol, triglycerides or the concentration of fat soluble vitamins. Sevelamer hydrochloride is a well tolerated alternative to calcium or aluminum containing phosphate binders and may offer an advantage to patients who become hypercalcemic on calcium-containing antacids and vitamin D supplementation. Furthermore, sevelamer hydrochloride lowers LDL cholesterol without affecting HDL cholesterol. The potential usefulness of the lipid lowering effects of sevelamer hydrochloride needs to be determined in additional prospective studies. PMID- 9877113 TI - Splenic lymphoma presenting as necrotizing glomerulonephritis. AB - The authors report a case of necrotizing glomerulonephritis, with granular immune deposits, leading to the etiologic diagnosis of splenic non-Hodgkin's Lymphoma (NHL). The serum creatinine, initially markedly elevated, decreased under steroids and chlorambucil. Unfortunately, the patient died from septic complications. The importance of a careful search for a (localized) NHL in case of necrotizing glomerulonephritis with granular immune deposits is emphasized. PMID- 9877114 TI - Disorders of water, electrolyte and acid-base balance in alcoholic patients. PMID- 9877115 TI - Slowing down chronic renal failure progression with various antihypertensive agents. PMID- 9877117 TI - Pathologic distinctions among the thrombotic microangiopathies. PMID- 9877116 TI - Long-term treatment of chronic hepatitis B with oral ganciclovir in two kidney transplant recipients. PMID- 9877118 TI - Urinary glycosaminoglycans in renal cell carcinoma. PMID- 9877119 TI - Emphysematous cystitis: possible side-effect of cyclophosphamide therapy. PMID- 9877121 TI - Lysosomal enzymes in preterm infants with bronchopulmonary dysplasia: a potential diagnostic marker. AB - Some lysosomal glycohydrolases (N-acetyl-beta-D-glucosaminidase and their major isoenzymes, beta-D-glucuronidase, alpha-D-galactosidase, beta-D-galactosidase and alpha-D-glucosidase) were investigated in the plasma of 36 preterm infants with respiratory distress, 11 of whom developed bronchopulmonary dysplasia (BPD), in order to evaluate the role of the lysosomal apparatus in the disease. Enzyme activity was assayed fluorimetrically; the major N-acetyl-beta-D-glucosaminidase (NAG) isoenzymes were separated using a routine chromatofocusing procedure; the diagnostic efficiency was evaluated by Bayes theorem. The mean levels of almost all glycohydrolases considered were significantly higher in BPD than in non-BPD infants. Among NAG major isoenzymes, an increase was found only in form A. No variation was evident in the plasma levels of glycohydrolases during dexamethasone therapy. Data from a retrospective analysis performed in all preterms considered, show that alpha-D-galactosidase and beta-D-galactosidase differentiate a posteriori BPD and non-BPD subjects. These enzymes, after a priori verification of their diagnostic potential in preterm infants at risk of BPD development, could acquire an important predictive value. PMID- 9877120 TI - A sensitive HPLC method for the quantification of free and total p-cresol in patients with chronic renal failure. AB - Para-cresol (4-methylphenol) is a volatile phenolic compound which is retained in chronic renal failure. Several recent studies suggest that p-cresol interferes with various biochemical and physiological functions at concentrations currently observed in uremia. Only a few methods are available for the determination of p cresol concentration in serum. In addition, these methods have only been used for the determination of total p-cresol. In particular, the evolution of free (non protein bound) p-cresol is of concern, because conceivably this is the biologically active fraction. The concentration of free p-cresol, is, however, markedly lower than that of total p-cresol, in view of its important protein binding. We report a method enabling the measurement of total and free p-cresol concentration in serum of healthy controls and uremic patients. Deproteinization, extraction and HPLC procedure are efficient, without interference of other protein bound ligands and/or precursors of p-cresol or phenol. By means of spiking experiments, the measurement of the UV absorbance over the 200-400 nm wavelength range, and capillary gas chromatography-mass spectrometry, the considered compound is identified as p-cresol. With a fluorescence detection at 284/310 nm as extinction/emission wavelengths the detection limit of p-cresol is 1.3 micromol/l (0.14 microg/ml). Recovery of added p-cresol to normal serum is 95.4+/-4.1%. For free p-cresol and total p-cresol determinations, intra-assay and day-to-day variation co-efficients are 3.2%, 4.2%, 6.9% and 7.3%, respectively. Compared to healthy controls, the serum p-cresol levels are 7-10 times higher in continuous ambulatory peritoneal dialysis patients (CAPD), uremic outpatients, and hemodialysis patients: 8.6+/-3.0 vs. 62.0+/-19.5, 87.8+/-31.7 and 88.7+/-49.3 micromol/l (0.93+/-0.32 vs. 6.70+/-2.11, 9.49+/-3.43, and 9.60+/-5.30 microg/ml) (p<0.05), respectively. The difference is even more important if free p-cresol is considered. This corresponds to a decreased protein binding in uremic patients. We conclude that the present method allows an accurate measurement of both total and free p-cresol, and that the measured concentrations in uremia are in the range which may cause biochemical alterations. PMID- 9877122 TI - Stabilities of N-acetyl-beta-D-glucosaminidase (NAG) isoenzymes in urine: advantage of NAG isoenzyme B measurement in clinical applications. AB - N-Acetyl-beta-D-glucosaminidase (NAG) is a widely used urinary enzyme for the assessment of renal diseases. We studied the stabilities of NAG isoenzymes in urine at 37 degrees C by enzyme assay and ELISA using a model simulating in vivo conditions. The stabilities were found to be affected by the pH. Under mild acidic condition (about pH 6), there was no significant loss of enzymatic activity of NAG isoenzyme A, enzymatic activity of NAG isoenzyme B and immunological activity of NAG isoenzyme B even after 8 h incubation. In contrast, under alkaline condition (about pH 8), the enzymatic activity of NAG isoenzyme A was rapidly lost, whereas both enzymatic and immunological activities of NAG isoenzyme B were maintained at more than 80% of their initial values. Also, we found that the ratios of endogenous NAG isoenzyme B to total NAG were elevated in alkaline urine samples. These results indicate that NAG isoenzyme A, which is a major isoenzyme in normal urine (pH 5-7), seems to be inactivated in alkaline urine. Our results suggest that for alkaline urine, NAG isoenzyme B should be measured to avoid misinterpretation of total NAG enzymatic activities. PMID- 9877123 TI - Effect of aminoguanidine on the generation of superoxide anion and nitric oxide by peripheral blood granulocytes of rats with streptozotocin-induced diabetes. AB - The effect of aminoguanidine (AG) on the production of superoxide anion O2*- and nitric oxide (NO) by peripheral blood granulocytes of rats with streptozotocin induced diabetes was studied. Induction of diabetes resulted in an increase of O2*-. Generation by both unstimulated and opsonized zymosan-stimulated granulocytes was maximal after 6 weeks and lower after 12 weeks. Treatment with aminoguanidine (1 g/l drinking water) decreased O2*- generation after 6 weeks but not after 12 weeks. NO production by both unstimulated and opsonized zymosan stimulated granulocytes was elevated in diabetic rats to a comparable extent after 6 and 12 weeks. AG attenuated this increased NO production. These results point to the beneficial effect of AG on oxidative stress in experimental diabetes and suggest an antioxidant effect of AG. PMID- 9877124 TI - Antioxidant vitamins and coronary artery disease risk in South African males. AB - Decreased antioxidant-vitamin nutritional status may increase lipid peroxidation and susceptibility of low-density lipoprotein (LDL) to oxidative modification. The aim of this study was to evaluate the vitamin nutritional status of coronary artery disease (CAD) patients and to assess the risk of CAD related to each individual antioxidant vitamin. The study was performed as a case-control study with 41 patients with angiographically demonstrated CAD and 41 apparently healthy age- and smoking status-matched controls. Plasma vitamin E, C and A concentrations were significantly decreased in CAD patients compared with controls (p < 0.001) after correcting for significant covariates. Per quartile decrease in vitamin A and E concentrations was associated with increased risk of CAD, even after adjusting for CAD risk factors, while per quartile decrease in vitamin C concentrations was not associated with significant CAD risk after adjusting for CAD risk factors. Decreased vitamin A and E concentrations are independently associated with increased risk of CAD independent from other CAD risk factors in white male South Africans and dietary intervention strategies are advocated. PMID- 9877125 TI - Beta 2-microglobulin does not behave as a biological marker of bone remodeling in patients with primary hyperparathyroidism. AB - Parathormone has been shown to increase the secretion and production of lysosomal enzymes including tartrate-resistant acid phosphatase. All our 68 patients with proven primary hyperparathyroidism had signs of hyperparathyroid bone disease. Tartrate-resistant acid phosphatase and bone alkaline phosphatase activities are raised as a result of enhanced bone remodelling. Serum beta 2-microglobulin concentration in patients with primary hyperparathyroidism was normal 1.6 (1.4 1.8) mg/l versus 1.8 (1.7-2.2) mg/l in 51 control subjects. In hyperparathyroid patients, serum beta 2-microglobulin concentration does not correlate with plasma tartrate-resistant acid phosphatase activity which is known to be a sensitive biological marker of bone remodelling (r = 0.088). Our findings indicate that serum beta 2-microglobulin does not behave as a biological marker of remodelling in patients with enhanced remodelling in primary hyperparathyroidism. PMID- 9877126 TI - Change in nitric oxide in humans due to application of a pneumatic tourniquet. AB - The nitric oxide profile produced by application of a pneumatic tourniquet was investigated in the plasma of 15 patients. Nitrite (NO2-) and nitrate (NO3-) plasma concentrations were measured simultaneously by high-performance liquid chromatography (HPLC). UV detection using the Griess reaction was done after the reduction of nitrate to nitrite. The plasma nitrate concentration 5 min after reperfusion was significantly higher than the concentrations before ischemia, immediately before reperfusion, and the next day. In contrast, there was no significant difference in the plasma nitrite concentrations before ischemia, immediately before reperfusion, 5 min after reperfusion, and the next day. These findings suggest that the generation of NO is important in ischemic reperfusion injury. PMID- 9877127 TI - Influence of feeding on metabolite excretion evidenced by urine 1H NMR spectral profiles: a comparison between subjects living in Rome and subjects living at arctic latitudes (Svaldbard). AB - Urines from 25 normal subjects living in Rome and 25 normal subjects living in Ny Alesund (Svaldbard) were analysed by 1HNMR spectroscopy. The observed differences in the concentration of the major metabolites were correlated to the composition of the diet. It was found that a diet rich of carbohydrates, such as the Italian diet, is responsible for an increased excretion of citrate, lactate, alanine, and glycine. Thus, a correct diagnostical interpretation of urinary metabolites needs to consider feeding habits. PMID- 9877128 TI - MRI of optic nerve and postchiasmal visual pathways and visual evoked potentials in secondary progressive multiple sclerosis. AB - We studied the relationship between abnormalities shown by MRI and functional disturbances in the visual pathway as assessed by the visual evoked potential (VEP) in 25 patients with established multiple sclerosis (MS); only 4 of whom had a history of acute optic neuritis. Optic nerve MRI was abnormal in 19 (76%) and is thus useful in detecting subclinical disease. Optic nerve total lesion length and area on the STIR sequence was found to correlate significantly with prolongation of the VEP latency. This may reflect a predominantly demyelinating rather than inflammatory origin for the signal change in the optic nerve. PMID- 9877129 TI - Serial MR observation of cortical laminar necrosis caused by brain infarction. AB - To examine the chronological changes characteristic of cortical laminar necrosis caused by brain infarction, 16 patients were repeatedly examined using T1-, T2 weighted spin-echo, T2*-weighted gradient echo, fluid attenuated inversion recovery (FLAIR) images, and contrast enhanced T1-weighted images at 1.0 or 1.5 T. High intensity cortical lesions were visible on the T1-weighted images from 2 weeks after ictus and became prominent at 1 to 3 months, then became less apparent, but occasionally remained at high intensity for 2 years. High intensity cortical lesions on FLAIR images became prominent from 1 month, and then became less prominent from 1 year, but occasionally remained at high intensity for 2 years. Subcortical lesions did not display high intensity on T1-weighted images at any stage. On FLAIR images, subcortical lesions initially showed slightly high intensity and then low intensity from 6 months due to encephalomalacia. Cortical lesions showed prominent contrast enhancement from 2 weeks to 3 months, but subcortical lesions were prominent from 2 weeks only up to 1 month. T2*-weighted images disclosed haemosiderin in 3 of 7 patients, but there was no correlation with cortical short T1 lesions. Cortical laminar necrosis showed characteristic chronological signal changes on T1-weighted images and FLAIR images. Cortical short T1 lesions were found not to be caused by haemorrhagic infarction. PMID- 9877130 TI - Differentiation of recent and old cerebral infarcts by diffusion-weighted MRI. AB - We performed MRI, including diffusion-weighted imaging, in 15 patients with recurrent strokes with acute ischaemia and at least one old lesion according to the clinical history and/or CT. Routine MRI showed similar signal intensity changes in both situations. Diffusion-weighted images, however, were positive in all acute or subacute infarcts. The high signal of acutely disturbed diffusion due to intracellular oedema could also be identified in small brain stem lesions. Spatial resolution was increased by applying separate gradients in each axis instead of creating anisotropy-independent trace images. PMID- 9877131 TI - Contrast-enhanced magnetisation transfer MRI in metastatic lesions of the brain. AB - Our purpose was to compare prospectively the sensitivity of contrast-enhanced magnetisation transfer (MT) MRI and gradient-echo (GE) T1-weighted images in metastatic disease of the brain. We studied 52 patients with brain metastases, using conventional T1-weighted GE and MT spin-echo (SE) images after the same standard dose of gadolinium. Axial 5-mm reconstructions of GE data were compared with 5-mm MT images in the same plane. Metastases were counted independently by two neuroradiologists. In 12 patients (23%) MT imaging showed more metastases than GE images (P = 0.03). We detected 68 more metastases with the former technique. PMID- 9877133 TI - High signal in the adenohypophysis on T1-weighted images presumably due to manganese deposits in patients on long-term parenteral nutrition. AB - Hypermanganesaemia is reported in patients on long-term parenteral nutrition. Deposition of manganese, giving high signal on T1-weighted images, may involve the basal ganglia. MRI in nine patients (mean age 51 years, range 31-75 years) on long-term parenteral nutrition (mean duration 30 months, range 6-126 months), demonstrated high signal in the anterior pituitary gland on T1-weighted sagittal and coronal images. The gland appeared normal on T2-weighted images. Signal intensity in the basal ganglia on T1-weighted images was increased in all patients. Endocrine assessment showed no significant abnormality. Neurological examination showed a mild parkinsonian movement disorder in one patient. Hypermanganaesemia was present in all nine (1.3-2.8 micromol/l, mean 1.87 micromol/l). The high signal in the anterior pituitary gland was probably related to deposition of paramagnetic substances, especially manganese. PMID- 9877132 TI - Sex differences in morphology of the brain stem and cerebellum with normal ageing. AB - The cerebral hemispheres become atrophic with age. The sex of the individual may affect this process. There are few studies of the effects of age and sex on the brain stem and cerebellum. We used MRI morphometry to study changes in these structures in 152 normal subjects over 40 years of age. In the linear measurements, men showed significant age-associated atrophy in the tegmentum and pretectum of the midbrain and the base of the pons. In women, only the pretectum of the midbrain showed significant ageing effects after the age of 50 years, and thereafter remained rather constant. Only men had significant age-associated reduction in area of the cerebellar vermis area after the age of 70 years. Both men and women showed supratentorial brain atrophy that progressed by decades. There were significant correlations between supratentorial brain atrophy and the diameter of the ventral midbrain, pretectum, and base of the pons in men, and between brain atrophy and the diameter of the fourth ventricle in women. PMID- 9877134 TI - Detection of an occult transclival cerebrospinal fluid fistula by CT and MRI. AB - We describe an unusual occult transclival cerebrospinal fluid (CSF) fistula to the sphenoid sinus demonstrated by MRI. CT was performed because of a posterior cerebral infarct caused by cardiac arrhythmia. Axial sections showed fluid in the sphenoid sinus. High-resolution scans revealed a bony defect 3 mm in diameter of the posterior wall of the sphenoid sinus, and MRI showed a transclival CSF fistula. This occult lesion was confirmed by surgery and duraplasty was successfully performed via an endonasal approach. PMID- 9877135 TI - The clinical significance of gadolinium enhancement of lumbar disc herniations and nerve roots on preoperative MRI. AB - The clinical significance of preoperative gadolinium DPTA enhancement around disc herniations and in the epidural space on MRI is not clear. The relation of nerve root enhancement to dysfunction also remains controversial. To investigate the clinical significance of contrast enhancement we looked at the symptoms and signs and gadolinium DPTA-enhanced images of 71 consecutive surgical candidates in a standardised fashion. Annular rupture and disc enhancement were found to be closely related, and correlated with sensory loss and paresis, respectively. Nerve root enhancement was found to be related to neurological deficits in general and sensory impairment in particular. PMID- 9877137 TI - An atretic parietal cephalocele associated with multiple intracranial and eye anomalies. AB - We present the cranial MRI findings in a 4-month-old girl with an atretic parietal cephalocele associated with multiple cerebral and ocular anomalies including lobar holoprosencephaly, a Dandy-Walker malformation, agenesis of the corpus callosum, grey-matter heterotopia, extra-axial cysts in various locations, bilateral microphthalmia and a retroocular cyst. PMID- 9877136 TI - Congenital muscular dystrophy with merosin deficiency: MRI findings in five patients. AB - We present the MRI findings in five patients with congenital muscular dystrophy (CMD) and merosin (laminin alpha2) deficiency, which was total in one and partial in four. In one patient with partial merosin deficiency, MRI was normal. The other four patients had supratentorial white matter abnormalities. In three, T2 weighted images revealed subcortical, deep lobar and periventricular high signal in white matter, while in the other there were only small peritrigonal areas of increased signal. On T1-weighted images, there was slightly low signal. Cortical abnormalities were absent. None of these changes were accompanied by symptoms or signs of central nervous system involvement. White matter abnormalities in a patient with CMD should prompt investigation of merosin. PMID- 9877138 TI - Aneurysmal malformation of the vein of Galen in three patients: clinical and radiological follow-up. AB - We studied three patients with aneurysmal malformations of the vein of Galen: all underwent embolisation followed by MRI and conventional angiography; two also underwent postembolisation magnetic resonance angiography (MRA). MRI was performed before treatment in two patients, MRA in only one, diagnosed prenatally. Two patients had normal appearances on follow-up MR studies and were completely cured by embolisation. However, the last patient, after two embolisations, despite the stabilisation of the clinical condition, underwent surgery and died. We think MRI is mandatory before endoarterial treatment, to assess the conditions of the brain. Angiography is mandatory only at the time of endovascular treatment, while MRA and MRI have a role in follow-up. Endoarterial embolisation remains the treatment of choice, and surgery is not advisable. PMID- 9877139 TI - Is local intra-arterial fibrinolysis contraindicated in elderly patients with cerebral artery occlusion? AB - Local intra-arterial fibrinolysis (LIF) is an effective treatment for selected patients in acute thromboembolic occlusion of the middle cerebral artery, vertebrobasilar system or ophthalmic artery. However, the risk/benefit ratio of thrombolysis in patient subgroups requires classification. Advanced age has been regarded as a prognostic factor for poor clinical outcome. We report our experience with LIF in seven patients with a mean age of 79 years (range 76-83 years) who represented thromboembolic occlusion of the basilar, middle cerebral or pericallosal arteries. We discuss why advanced age need not contraindicate LIF. PMID- 9877140 TI - Award of the Fourth Lucien Appel Prize for Neuroradiology. PMID- 9877142 TI - Cycle rate variations in roller ski skating: effects on oxygen uptake and poling forces. AB - The purpose of this study was to examine the effect of cycle rate (CR) variations on the metabolic cost and upper body forces during roller skiing with the V2 alternate technique on flat terrain. Nine highly skilled cross-country skiers roller skied at a paced speed of 18.0+/-0.1 km x h(-1) using their chosen CR, and CRs that were 10% slower and 20% faster. Oxygen uptake (VO2) was determined through collection of expired gases into a meterological balloon and poling forces were measured with piezoelectric transducers during the last 30 s of each four minute trial of roller skiing. One-way repeated measures ANOVA revealed that VO2 varied significantly with CR (p=0.02) with the chosen CR being significantly lower than the higher CR (p < 0.05). Poling forces and poling time were not significantly different among the CR conditions. The present results demonstrate that 1) an alteration in cycle rate affects metabolic cost of roller ski skating, 2) skiers tend to naturally select the most economical cycle rate, and 3) moderate variations in cycle rate do not appear to affect propulsive force generation through the poles in roller skiing. PMID- 9877141 TI - Stretch-shortening cycle in roller ski skating: effects of technique. AB - The purposes of this study were 1) to examine the occurrence of the stretch shortening cycle (SSC) in the three principal techniques of ski skating and 2) to test the hypothesis that differences in SSC behaviour may explain the choice of a technique when the grade is altered. Ten highly skilled male cross-country skiers roller skied at 5.3+/-0.1 m x s(-1) using the V1 skate (V1), V2-alternate (V2A), and V2 skate (V2) techniques. Knee and ankle joint kinematics, EMG of the vastus lateralis (VL) and gastrocnemius lateralis (GL) muscles, and roller ski velocity were recorded during the last 40 s of each bout of roller skiing. The existence of an EMG signal during the eccentric phase prior to the thrust, as well as the lack of significant delay between the two phases, showed that SSC occurs in the three techniques of roller ski skating with the exception of 50% of the subjects at the weak side knee with V1. The VL stretching velocity over the eccentric phase was higher (p < 0.005) with V2 and V2A than V1, indicating that less elastic energy was stored in the lower limbs extensor muscles with V1. This could partly explain why this technique is not used on flat terrains during races. However, no differences of average EMG activity or IEMG existed between techniques for either GL or VL during the concentric phase, suggesting that the theoretical advantage due to the SSC behaviour is minor in roller ski skating. PMID- 9877144 TI - Exhaustive endurance training for 6-9 weeks did not induce changes in intrinsic heart rate and cardiac autonomic modulation in female athletes. AB - We investigated the effects of progressively increased training load and overtraining on resting and intrinsic heart rate (IHR) and cardiac autonomic modulation (CAM), and their relationships to performance variables. Nine athletes (ETG) increased training volume at 70-90% of maximal oxygen uptake (VO2max) by 130% (p<0.01) and training volume at <70% VO2max by 100% (p < 0.01) during 6-9 weeks. The corresponding increases in six female control athletes (CG) were 5 and 10%. Pharmacological blocking through atropine and propranolol and the Rosenblueth and Simeone model were used to calculate the sympathovagal balance index (Abal) and to measure IHR. The results were analysed using two-way analysis of variance. VO2max, IHR and Abal did not change. Resting heart rate had a tendency to decrease in the ETG and increase in the CG during the training period (interaction p < 0.01). Five ETG athletes demonstrated overtraining state (OA subgroup). Their VO2max (mean+/-SEM) decreased from 53.0+/-2.2 ml x kg(-1) x min( 1) to 50.2+/-2.3 ml x kg(-1) x min(-1) (p < 0.01), but no changes in resting HR, IHR and Abal were found. A significant correlation between the baseline values of VO2max and the parasympathetic activity index was found (r=-0.59, p < 0.05). In conclusion, progressively increased training load and overtraining did not induce significant changes in intrinsic heart rate or cardiac autonomic modulation in female endurance athletes. Resting heart rate rather decreased with heavy endurance training and overtraining. High maximal oxygen uptake was correlated with high cardiac parasympathetic modulation. PMID- 9877143 TI - Muscle strength and hormonal levels in adolescents: gender related differences. AB - The purpose of the present investigation was to study muscle strength in adolescents and its relationship to serum levels of testosterone and growth hormone in both genders. Thirty active adolescents (15 boys; age range 11 -12 y/o) participated in the first study. Isokinetic muscle strength of the dominant knee extensors (KE) was determined at 0, 12, 20, 30, 120, 180 and 240 deg/sec using a Cybex 340 dynamometer. The assessment of pubertal status was accomplished using the criteria of Tanner. Serum levels of total testosterone (T) and growth hormone (GH) were determined using radioimmunoassay techniques. Boys had higher (p< 0.001) T levels but no differences in muscle strength were detected between genders. Fifty-seven additional subjects representing three age groups (11-12 y/o, n=18; 13-14, n=21; 17-18, n=18) participated in the second study. A significant increase in peak torque (absolute and corrected for body weight) with age was observed in both genders. There were no significant gender differences in strength for the two youngest age groups, but boys were stronger than girls in the oldest age group (group 3). Testosterone and GH levels increased with age in boys but not in girls. Gender related differences in T were found in groups 2 and 3. A positive correlation (r=0,64 boys; r=0.46 girls) between testosterone levels and absolute muscle strength was seen in both genders. Our results suggest that increases in anabolic hormones precede muscle strength gains in adolescent males. In addition, gender related differences in muscle strength during adolescents cannot be explained solely on the basis of difference in body size or T levels. PMID- 9877145 TI - Relationship between muscle fiber pennation and force generation capability in Olympic athletes. AB - The thickness (TBmt) and fiber pennation angle (TBpen) of triceps brachii as well as isokinetic force developed during elbow extension were measured in Olympic athletes to investigate the relationship between muscle fiber pennation and force generation capability. The subjects were male members of the 1996 Japanese Olympic team who competed in seven different events; 9 wrestlers, 16 soccer players, 11 sprinters, 5 judo athletes, 7 gymnasts, 9 rowers and 18 baseball players. The TBmt and TBpen, measured by a B-mode ultrasound, ranged between 29 mm and 50 mm and between 11 degrees and 30 degrees, respectively, and on average were larger in the judo athletes, wrestlers and gymnasts compared to the other groups. A significant correlation (r=0.580, p < 0.05) was found between TPpen and TBmt per unit of the upper arm length, and so the observed event-related differences in TBpen tended to reflect the differences in TBmt. The isokinetic forces relative to the cross-sectional area (CSA) estimated from TBmt, measured at two constant velocities of 1.05 rad/s (F1.05/CSA) and 3.14 rad/s (F3.14/ CSA), were negatively correlated to the CSA; r=-0.617 (p < 0.05) for F1.05/CSA and r= 0.635 (p < 0.05) for F3.14/CSA. In addition, low but significant negative correlations existed between TBpen and both F1.05/CSA (r=-0.365, p < 0.05) and F3.14/ CSA (r=-0.336, p <0.05). Even when the effect of TBpen was statistically normalized, the F1.05/CSA and F3.14/CSA were still negatively correlated to the CSA, r=-0.530 (p < 0.05) for F1.05/ CSA and r=-0.561 (p < 0.05) for F3.14/CSA. Therefore, at least in the Olympic athletes tested in this study, the magnitude of the pennation angles reflects muscle size, but it does not seem to be a factor that explains extensively the lower F/CSA in athletes with large muscle size. PMID- 9877146 TI - The effect of exercising to exhaustion at different intensities on saliva immunoglobulin A, protein and electrolyte secretion. AB - The quality and quantity of saliva may be important in defending against pathogens transmitted via the buccal cavity. The aim of the present study was to examine the effect of cycling to exhaustion at moderate and high intensity on various salivary parameters and the time course of recovery. Eighteen male subjects of mixed physical fitness took part in the study. Subjects performed two bouts of exercise on separate occasions at least one week apart. Following an overnight fast, subjects cycled on an electrically braked cycle ergometer at a work rate equivalent to 80% VO2max until exhaustion. On another occasion they cycled on the same ergometer at 55% VO2max for 3 h or to fatigue (whichever was sooner). The order of the rides was randomised. Timed, unstimulated saliva samples were collected pre-exercise, during exercise, at cessation of exercise and at 1, 2.5, 5 and 24 h post-exercise. Saliva samples were analysed for IgA, total protein and osmolality. Saliva flow rate was significantly reduced by exercise (P < 0.01). Saliva IgA concentration, secretion rate and ratio to osmolality increased during exercise (P < 0.01). IgA to protein ratio did not change significantly during exercise. Since saliva protein secretion rate increased during exercise (P < 0.01) it appears that correcting for loss of saliva water by expressing IgA relative to protein is misleading. IgA secretion rate and IgA to osmolality ratio are more appropriate measures and neither parameter was lowered by exercise. The results of this study indicate that exercise may detrimentally affect the quantity of saliva produced, but not the quality of saliva. Furthermore, when exercise is to exhaustion, the intensity of the bout does not appear to influence the saliva response. Neither exercise protocol had any long term effect on saliva as all variables recovered within 1 h post-exercise. PMID- 9877147 TI - The Conconi test: a controversial concept for the determination of the anaerobic threshold in young rowers. AB - In order to examine the validity of Conconi's heart rate threshold (ATHR) during rowing ergometry, in accordance with the new recommendations of Conconi, ten young rowers (age 17.5 [SD 1.7] years; height 182.5 [SD 5.9]cm; body mass 77.0 [SD 10.6] kg) were submitted - at random and within 48-72 h - to two progressive incremental exercise tests on a rowing ergometer. In the first test, the heart rate threshold (ATHR) was determined during a ramp-like incremental test. In a second test, the lactate method (150W + 50W x 3 min(-1)) was used to determine the 4 mmol x l(-1) threshold (AT4) and the individual anaerobic threshold (IAT). Subsequently, each subject performed within 48-72 h after the second progressive incremental exercise test a 30 min prolonged exercise test (PET) at the power output corresponding to ATHR to demonstrate whether or not a steady state in blood lactate concentration ([La-]bSS) could be maintained. Friedmann's analysis of variance, Wilcoxon signed ranks test and Spearman rank correlation coefficient (rs) were used as statistical tools. Significant lower values for power output (p < 0.01 ) and heart rate (HR) (p < 0.01) were found at IAT as compared to AT4 and ATHR. No relationships were found between ATHR on the one hand and AT4 and IATon the other hand, neither for power output (rs=0.27 and -0.09), nor for HR (rs=0.08 and -0.08). Coefficients of correlation between AT4 and IAT were respectively 0.68 (p < 0.05) for power output and 0.79 (p < 0.01) for HR. During PET only four out of 10 rowers reached the target time of 30 min at ATHR power output and only one rower showed a lactate steady state. From our results we can conclude that power output at ATHR is overestimated and cannot be used for the prescription of endurance training in rowing. The ATHR does not reflect the anaerobic threshold and is therefore not relevant for monitoring continuous endurance training in rowing. PMID- 9877148 TI - Reliability and concurrent validity of Futrex and bioelectrical impedance. AB - Thirty Caucasian males (aged 19-32yr) participated in this study designed to investigate the reliability of multiple bioelectrical impedance analysis (BIA) and near-infrared spectroscopy (Futrex, FTX) measurements and the validity of BIA and FTX estimations of hydrostatically (UW) determined percent body fat (%BF). Two BIA and two FTX instruments were used to make 6 measurements each of resistance (R) and optical density (OD) respectively over a 30 min period on two consecutive days. Repeated measures ANOVA indicated that FTX and BIA, using manufacturer's equations, significantly (p<0.01) under predicted UW by 2.4 and 3.8%BF respectively. Standard error of estimate (SEE) and total error (TE) terms provided by regression analysis for FTX (4.6 and 5.31%BF respectively) and BIA (5.65 and 6.95%BF, respectively) were high. Dependent t-tests revealed no significant differences in either FTX or BIA predictions of %BF using two machines. Intraclass reliabilities for BIA and FTX estimates of UW %BF across trials, days, and machines all exceeded 0.97. A significant random error term associated with FTX and a significant subject-by-day interaction associated with BIA was revealed using the generalizability model. Although FTX and BIA estimates of UW %BF were reliable, due to the significant underestimation of UW %BF and high SEE and TE, neither FTX nor BIA were considered valid estimates of hydrostatically determined %BF. PMID- 9877149 TI - Inertial effects on the assessment of performance in isokinetic dynamometry. AB - Moments due to acceleration and deceleration periods influence the joint moment (torque) in isokinetic dynamometry. The purpose of this study was to evaluate the differences in the peak moment and the moment development during the initial acceleration period of isokinetic concentric knee extension before and after correction for inertial effects, and evaluate their relationship. The moment development was expressed by the work produced during the initial 0.125 s of the movement. Five male and five female healthy young adults participated in the study. Seven concentric knee extensions at 1.04, 3.14 and 5.24 rad/s were performed on a Biodex dynamometer. Inertial moments were calculated using acceleration data derived from the Biodex software angular displacement data. After inertial correction, the maximum errors in the original peak moment and the moment development at the inital 0.125 s were 7 and 16%, respectively, and were recorded at 5.24rad/s. At slower velocities (1.04 and 3.14 rad/s) the effects of inertial moments on the peak moment and the work at the initial 0.125 s were negligible. Inertial moments did not influence the relationship between the examined parameters at the three velocities. In conclusion, inertial moments affect the peak moment and moment development during the initial acceleration period at high velocities of isokinetic testing and therefore these effects should be considered in order to obtain valid results in isokinetic dynamometry. PMID- 9877150 TI - Carbohydrate supplementation and the lymphocyte proliferative response to long endurance running. AB - This randomized, double-blind, placebo-controlled study examined the influence of 6% carbohydrate ingestion on hormonal and lymphocyte proliferative responses (5 total samples over 9 hours) to 2.5 h of high-intensity running by 30 experienced marathon runners. The T-cell response differed between groups, with the placebo group exhibiting a greater increase immediately post-run and greater decrease at 3 h of recovery. No group differences were observed for Con A-, PHA-, or PWM induced lymphocyte proliferation. However, when PHA was adjusted per T-cell, group differences were observed, highlighted by a decrease in the placebo group immediately post-run. Glucose and cortisol responses differed between groups, with glucose lower and cortisol higher in the placebo group immediately post-run. Post-run glucose correlated negatively with postrun cortisol (r=-0.670, P< 0.001) and epinephrine (r=-0.540, P=0.002). Post-run cortisol also correlated negatively with total lymphocytes and T-cells at 1.5 hours (r=-0.429, P=0.018 and r=-0.424, P=0.019, respectively) and 3 hours (r=-0.566, P=0.001 and r=-0.523, P=0.003, respectively) of recovery. The pre- to post-run change in glucose correlated to the same changes in PHA/T-cell (r=0.456, P=0.011). The data support an interactive effect of carbohydrate ingestion on plasma glucose and cortisol. The data support an interactive effect of carbohydrate ingestion on plasma glucose and cortisol, T-cell trafficking, and cell-adjusted PHA-induced lymphocyte proliferation following long endurance running. PMID- 9877151 TI - Influence of one bout of intensive running on lymphocyte micronucleus frequencies in endurance-trained and untrained men. AB - Exercise induced chromosomal damage was evaluated in trained and untrained subjects, who performed treadmill running at 85% of maximal oxygen uptake for 30 min. The subjects had their peripheral blood taken before, immediately after and 30 min after the running test for the analysis of lymphocyte chromosomal damage that was evaluated by micronucleus assay. The blood samples were also subjected to X-ray irradiation in vitro to examine the modification of exercise induced chromosomal damage by a secondarily induced oxidative stress. Spontaneous chromosomal damage in lymphocytes did not significantly increase at least until 30 min after the running both in the trained and untrained subjects. However, the X-ray-induced chromosomal damage was significantly enhanced at 30 min after the running in the untrained group, but not in the trained group. The ratio of X-ray induced/spontaneous chromosomal damage also tended to increase after the running only in the untrained group. These preliminary results suggest that intensive exercise induced very slight chromosomal damage only in the untrained group, which could be intensified by the secondarily induced oxidative stress. PMID- 9877152 TI - Physical fitness and sports skills in relation to sports injuries. A four-year prospective investigation of sports injuries among physical education students. AB - In order to study the relationship between physical fitness/sport-specific skills and sports injuries 136 physical education students were studied during their 4 years of training in a prospective investigation. Physical fitness was measured every year using a battery of fitness tests, and the performance marks of a number of sports scored at the exams of the academy were used as parameters for the sport-specific skills. Sports injuries were recorded every 3 weeks on standard forms. Relative risk ratios were calculated between the tertile groups good, average and poor for all variables of physical fitness and sport-specific skills. Injury-proneness was defined for all and for acute and chronic injuries separately near the median number of injuries sustained. In only 6 out of 126 computed relative risks was a significant difference found. Discriminant analysis revealed an explanation of 16%, 14% and 11% of the variance for respectively all, acute and chronic injuries, at which 5 or 6 variables in varying combination were included. From our findings it may be concluded that physical fitness and sport specific skills have little impact on sports injuries for the following two main reasons. Firstly, subjects at risk for sports injuries participate per definition in sports activities and have consequently developed their fitness and skills compared to the sedentary population. Thus, the range in physical fitness or sports skills in the population at risk is relatively small (physical education students belong to the 7th-10th decile in fitness test scores within a general college student population) and therefore an effect is hard to show. Secondly, the total number of sports injuries is very small and moreover, it should be distributed over several categories for analysis. The favourable advantages of using physical education students to study intrinsic risk factors (comparable and varied sports program, excellent compliance) appeared to be insufficient to compensate for drawbacks of selection. PMID- 9877153 TI - Unregulated exposure of the ribosomal M-site caused by NAC depletion results in delivery of non-secretory polypeptides to the Sec61 complex. AB - Nascent polypeptide associated complex (NAC) interacts with nascent polypeptides emerging from ribosomes. Both signal recognition particle (SRP) and NAC work together to ensure specificity in co-translational targeting by competing for binding to the ribosomal membrane attachment site. While SRP selects signal containing ribosomes for targeting, NAC prevents targeting of signal peptide-less nascent chains to the endoplasmic reticulum membrane. Here we show that the ribosome binding that occurs in NAC's absence delivers signalless nascent chains to the Sec61 complex, underscoring the danger of unregulated exposure of the ribosomal M-site. Recently, the idea that NAC prevents ribosome binding has been challenged. By carefully examining the physiologic NAC concentration in a variety of tissues from different species we here demonstrate that the discrepancy resulted from subphysiologic NAC concentrations. PMID- 9877155 TI - Sulfur K-edge X-ray absorption spectroscopy for determining the chemical speciation of sulfur in biological systems. AB - Sulfur is an essential biological element, yet its biochemistry is only partially understood because there are so few tools for studying this element in biological systems. X-ray absorption spectroscopy provides a unique approach to determining the chemical speciation of sulfur in intact biological samples. Different biologically relevant sulfur compounds show distinctly different sulfur K-edge X ray absorption spectra, and we show here, as an example, that this allows the deconvolution of the sulfur species in equine blood. PMID- 9877154 TI - The CGRP receptor can couple via pertussis toxin sensitive and insensitive G proteins. AB - Swiss 3T3 cell lines were constructed co-expressing receptor activity modifying protein (RAMP) 1 with the calcitonin receptor-like receptor (CRLR), and showed 125I-calcitonin-gene-related peptide (CGRP) 1 binding indicative of a type I CGRP receptor. Application of CGRP1 led to an increase in cAMP, which in 2/5 cell lines was augmented following pertussis toxin (PTX) pre-treatment. In Xenopus oocytes, expression of RAMP1, which potentiates an endogenous CGRP receptor, led to constitutive activation of co-expressed GIRK potassium channels. This potassium current was increased following CGRP application or co-expression of CRLR, but decreased by PTX or co-expression of transducin. We conclude that the CGRP receptor can signal to both PTX sensitive and insensitive G proteins. PMID- 9877156 TI - Regulation of System A amino acid transport in L6 rat skeletal muscle cells by insulin, chemical and hyperthermic stress. AB - In this study we have investigated the effects of insulin, chemical and hyperthermic stresses upon the activity of the System A amino acid transporter in L6 rat muscle cells. Uptake of alpha-methyl-aminoisobutyric acid (Me-AIB), a non metabolisable System A substrate, was increased by between 50% and 80% when muscle cells were exposed to a maximally effective concentration of insulin (100 nM), sodium arsenite (ARS, 0.5 mM) or a 42 degrees C heat shock (HS). The observed activation in System A in response to all three stimuli was maximal within 20 min and in the case of insulin and ARS primarily involved an increase in the Vmax of System A transport. In contrast, HS induced significant increases in both Vmax and Km of System A transport suggesting that hyperthermic stress may activate System A by a mechanism distinct from that mediating the effects of insulin and ARS. The hormonal stimulation of System A was blocked by the phosphoinositide 3-kinase (PI3k) inhibitor, wortmannin, but not by rapamycin or PD 98059 which respectively inhibit the mTOR and classical MAP kinase pathways. Exposure of L6 cells to ARS, but not HS, caused a 4.7-fold stimulation in MAPKAP K2 activity that was blocked by SB 203580, a specific inhibitor of the stress activated protein kinase SAPK2/p38. However, neither SB 203580, rapamycin nor wortmannin were able to suppress the ARS- or HS-induced stimulation in System A transport. In summary, our results demonstrate that activity of the System A transporter can be rapidly upregulated in response to hormonal and stress stimuli through changes in the transport kinetics of the System A carrier. Our data show that whilst the hormonal response is PI3k dependent, the signalling mechanisms which instigate changes in System A activity in response to chemical or hyperthermic stress do not appear to involve PI3k or components of the mTOR, p42/p44 MAP kinase or SAPK2/p38 signalling pathways. PMID- 9877157 TI - GD1alpha-replica peptides functionally mimic GD1alpha, an adhesion molecule of metastatic tumor cells, and suppress the tumor metastasis. AB - A novel peptide technology to produce mimicking peptides of carbohydrate moiety (which we propose to name glyco-replica peptides) is a useful tool to elucidate the functions of glycoconjugate. Carbohydrate moiety of ganglioside GD1alpha functions as a molecule involved in the adhesion between murine highly metastatic lymphoma RAW117-H10 cells and hepatic sinusoidal endothelial (HSE) cells. To prepare peptides which mimic the carbohydrate structure of GD1alpha, phage clones expressing peptides which bound to a monoclonal antibody against GD1alpha (KA17) were isolated from a phage-displayed random peptide library. Four phage clones having affinity to the monoclonal antibody KA17 were isolated, and these clones showed inhibitory effect on the binding of KA17 to GD1alpha. The amino acid sequences of the displayed pentadecamers were determined, and one of the phages displaying sequence WHWRHRIPLQLAAGR bound to HSE cells directly and showed the highest inhibitory effect on the adhesion between RAW117-H10 cells and HSE cells. The synthesized peptides having the same sequences to the displayed 15mers in the four isolated phage clones also showed the inhibitory effect on the adhesion of RAW117-H10 cells to HSE cells, and, again, the WHWRHRIPLQLAAGR peptide showed the highest inhibitory effect. Furthermore, intravenous injection of the peptide brought almost complete inhibition of the metastasis of RAW117-H10 cells to lung and spleen, and about 50% inhibition of the liver metastasis. These results indicate that GD1alpha plays an important role for metastasis of RAW117-H10 cells, and the peptides obtained by the present procedure are able to mimic the functional role of the glycoconjugate. PMID- 9877159 TI - Biochemical evidence that the N-terminal segments of the alpha subunit and the beta subunit play interchangeable roles in the activation of protein kinase CK2. AB - The concept that the amino-terminal segment plays a role in conferring high basal activity to protein kinase CK2 alpha subunit has been validated by generating two mutants (Y26F and delta2-6) which are defective both in catalytic activity and in thermal stability. The additional finding that the activity of the two mutants is fully restored upon association with the regulatory beta subunit, in conjunction with the observation that synthetic peptides reproducing the N-terminal segment (1-30) and the activation loop (175-201) of CK2alpha counteract the functional effects of the C-terminal domain of the beta subunit, is consistent with a mechanism of activation of CK2 where the N-terminal domain of alpha and the C terminal domain of beta play interchangeable roles. PMID- 9877158 TI - Selective inhibition of human type 1 11beta-hydroxysteroid dehydrogenase by synthetic steroids and xenobiotics. AB - Functional analyses were performed with microsomal human 11beta-hydroxysteroid dehydrogenase type 1 overexpressed in the yeast Pichia pastoris. Cell extracts or microsomes from transformed strains displayed dehydrogenase and reductase activities, which were up to 10 times higher than in human liver microsomes, while for whole cells cortisone reduction but no dehydrogenase activity was observed. The synthetic glucocorticoids prednisolone and prednisone were efficiently metabolized by subcellular fractions, whereas no activity was observed with dexamethasone, budesonide and deflazacort. Inhibitors found to be effective towards the recombinant 11beta-hydroxysteroid dehydrogenase include synthetic steroids and xenobiotic compounds, revealing selective inhibition of the reaction direction, useful for development of specific inhibitors. PMID- 9877160 TI - Isolated endosomes from quiescent rat liver contain the signal transduction machinery. Differential distribution of activated Raf-1 and Mek in the endocytic compartment. AB - In this study we identify the molecules involved in the MAPK signal transduction pathway (Ras, Raf-1, Mek, Mek-P and MAPK) in highly purified endosomal fractions isolated from rat liver. Biochemical analysis shows that only the early-sorting endocytic compartment contains activated Raf-1 and Mek. Finally, the exogenous administration of EGF led to redistribution of Raf-1 from the caveolin-enriched plasma membrane into the endosomes. PMID- 9877161 TI - Successful transfer of ADA gene in vitro into human peripheral blood CD34+ cells by transfecting EBV-based episomal vectors. AB - We report a novel non-viral system for transfecting human immature hematopoietic cells in vitro. Epstein-Barr virus (EBV)-based episomal vectors carrying human adenosine deaminase (ADA) gene cDNA were transfected by electroporation into human peripheral blood (PB) CD34+ cells. The transgene-specific mRNA were detected from 37 to 100% of CFU-c (colony forming unit in culture) colonies derived from the transfected cells. A two-fold increase in enzyme activity was also found. These results indicate the successful transfer and expression of genes in human immature hematopoietic cells using the EBV-based episomal vector system. PMID- 9877163 TI - Lipid composition of Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn) insect cells used for baculovirus infection. AB - The lipid composition of two different insect cell lines from Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn) which are established cell lines for infection with recombinant baculovirus was analyzed by high-performance liquid chromatography and gas-liquid chromatography. The major phospholipids found were phosphatidylcholine and phosphatidylethanolamine, the major mono-unsaturated fatty acids were oleic acid and palmitoleic acid, the major saturated fatty acid was stearic acid. The cholesterol to phospholipid ratio was demonstrated to be lower than in mammalian cell lines. Infection with a recombinant baculovirus Autographa californica resulted in increased levels of phosphatidylcholine in the insect cells. The baculovirus/insect cell system has become a popular system for heterologous protein production. Functional changes of membrane proteins produced in these two cell lines might be correlated to a different lipid profile of their cellular membranes. PMID- 9877162 TI - Napsins: new human aspartic proteinases. Distinction between two closely related genes. AB - cDNA sequences were elucidated for two closely related human genes which encode the precursors of two hitherto unknown aspartic proteinases. The (pro)napsin A gene is expressed predominantly in lung and kidney and its translation product is predicted to be a fully functional, glycosylated aspartic proteinase (precursor) containing an RGD motif and an additional 18 residues at its C-terminus. The (pro)napsin B gene is transcribed exclusively in cells related to the immune system but lacks an in-frame stop codon and contains a number of polymorphisms, one of which replaces a catalytically crucial Gly residue with an Arg. Consideration is given to whether (pro)napsin B may be a transcribed pseudogene or whether its putative protein product undergoes rapid intracellular degradation. PMID- 9877164 TI - Production and characterization of monoclonal antibodies directed against native epitopes of NhaA, the Na+/H+ antiporter of Escherichia coli. AB - Monoclonal antibodies (mAbs) recognizing native epitopes are an important tool for functional and structural studies of proteins, yet they have rarely been used with transport proteins. In an attempt to raise monoclonal antibodies against the NhaA Na+/H+ antiporter of Escherichia coli we encountered difficulties in the screening procedure, which is based on the standard enzyme-linked immunosorbent assay (ELISA). Here we report a rapid and efficient method of screening for anti NhaA mAbs which recognize native epitopes of the antiporter. The method is based on the use of His-tagged protein, Ni2+-nitrilotriacetic acid coated plates and non-denaturing conditions in the assay. With this procedure four mAbs were obtained, three of which recognize the NhaA in its native conformation and one preferentially recognizes the denatured form. The latter mAb is Western blot positive, the others are Western blot negative and bind the detergent solubilized NhaA as assayed by gel filtration chromatography. Competition experiments show that the native epitopes are common to both the His-tagged and the wild-type protein. We suggest that in the standard ELISA the NhaA protein is not presented to the antibody in the native conformation whereas the His tag based protocol favors this presentation. Indeed, we could remarkably improve the low reactivity of the standard ELISA by coating the plates with anti-NhaA mAb and use it to present NhaA ('sandwich' ELISA or two antibodies assay). Remarkably, two of the mAbs (5H4, 2C5) which bind native NhaA inhibit drastically the deltapH driven 22Na uptake mediated by His-tagged NhaA reconstituted in proteoliposomes. Hence, these mAbs afford a new tool to study the structure/function relationship of the antiporter. PMID- 9877165 TI - A protein import receptor in pea chloroplasts, Toc86, is only a proteolytic fragment of a larger polypeptide. AB - The protein import complex of the chloroplastic outer envelope (Toc-complex) contains a prominent subunit of 86 kDa molecular weight (Toc86). Toc86 was identified as a putative precursor receptor. The Arabidopsis genome sequencing project indicates that Toc86 represents only a proteolytic fragment of a larger polypeptide of 160 kDa. The 160-kDa protein, which we name Toc160, is only present in significant amounts in pea chloroplasts isolated under stringent conditions. The capacity of chloroplasts to import an in vitro translated precursor protein correlates well with the integrity of Toc160. We conclude that Toc160 is still a bonafide subunit of the protein import machinery of chloroplasts. PMID- 9877166 TI - Lysophosphatidylcholine generates superoxide anions through activation of phosphatidylinositol 3-kinase in human neutrophils. AB - Lysophosphatidylcholine (LPC) accumulates in inflammatory tissues, where neutrophils are recruited to generate superoxide anions (O2.-). Here, we show that LPC stimulates O2.- generation in human neutrophils and that the activity is inhibited with phosphatidylinositol 3-kinase (PI3 kinase) inhibitors, but not with protein kinase C (PKC) inhibitors. Furthermore, we demonstrate that LPC activates PI3 kinase in neutrophils. Thus, LPC might contribute to host defense by generating O2.- in neutrophils through PI3 kinase activation, but not through PKC activation. PMID- 9877167 TI - Chimeric Galpha(q) mutants harboring the last five carboxy-terminal residues of Galpha(i2) or Galpha(o) are resistant to pertussis toxin-catalyzed ADP ribosylation. AB - Three widely-used Galpha(q) chimeras harboring the last five residues of Galpha(i), Galpha(o) and Galpha(z) (qi5, qo5 and qz5) were examined for their ability to serve as substrates for pertussis toxin (PTX)-catalyzed ADP ribosylation. In COS-7 cells coexpressing one of the three opioid receptors (mu, delta, and kappa) and a Galpha(q) chimera, agonist-induced stimulation of phosphoinositide-specific phospholipase C (PI-PLC) was largely insensitive to PTX treatment. Only the qi5-mediated stimulation of PI-PLC by kappa-opioids was partially inhibited by PTX. In betagamma-release assays, PTX treatment did not affect the ability of opioid receptors to activate these chimeras. [32P]ADP ribosylation labeled Galpha(i/o) but not qi5 or qo5, although the expression of these chimeras was confirmed by immunodetection. Thus, Galpha(q) chimeras with a Galpha(i/o)-like tail are insensitive to PTX treatment. PMID- 9877168 TI - Induction of a spectroscopically defined transition by guanidinium hydrochloride on a recombinant calcium binding protein from Entamoeba histolytica. AB - Sequence analysis and metal ion binding studies reported earlier have established that the calcium binding protein (CaBP) from the parasitic ameboid Entamoeba histolytica protein has four canonical EF hand motifs which bind calcium. Equilibrium denaturation studies on both the apo and the holo forms of this protein indicate the presence of stable transition intermediates at low denaturant concentrations as revealed by the binding of the non-specific hydrophobic dye ANS. Fast reaction kinetics shows that the binding of the Gdn+ ions at or near the Ca2+ sites in the N-terminal domain influences metal ion binding to the sites in the C-terminal domain. Isothermal calorimetric titrations performed using low GdnHCl concentrations reveal the presence of two binding sites of low affinity, both being endothermic in nature. Thus the stabilization of CaBP observed at low GdnHCl concentration represents a native-like intermediate, with the Gdn+ ions mimicking Ca2+ binding at the N-terminal domain of this protein. PMID- 9877169 TI - Accessibility of selenomethionine proteins by total chemical synthesis: structural studies of human herpesvirus-8 MIP-II. AB - The determination of high resolution three-dimensional structures by X-ray crystallography or nuclear magnetic resonance (NMR) is a time-consuming process. Here we describe an approach to circumvent the cloning and expression of a recombinant protein as well as screening for heavy atom derivatives. The selenomethionine-modified chemokine macrophage inflammatory protein-II (MIP-II) from human herpesvirus-8 has been produced by total chemical synthesis, crystallized, and characterized by NMR. The protein has a secondary structure typical of other chemokines and forms a monomer in solution. These results indicate that total chemical synthesis can be used to accelerate the determination of three-dimensional structures of new proteins identified in genome programs. PMID- 9877170 TI - Photochemical localization of the semotiadil binding region within the cardiac Ca2+ channel alpha1 subunit. Comparison with the skeletal muscle counterpart. AB - We have previously identified the binding region of a new Ca2+ antagonist semotiadil in the skeletal muscle Ca2+ channel. To the same semotiadil derivatives, the cardiac counterpart showed distinct and different binding characteristics: semotiadil and its photoaffinity analog D51-4700 inhibited [3H]PN200-110 binding to cardiac membrane preparations with IC50 values of 13-20 microM, which are 10 times higher than those in skeletal muscle. Hill slopes of the binding inhibition were 0.94-1.0 for the cardiac channels compared to 0.63 0.67 for the skeletal muscle channels. A possible explanation for the difference is that the semotiadil binding site is differently conferred in cardiac and skeletal muscle Ca2+ channels. To reveal this within the primary structure, photoaffinity labeling of cardiac membranes was employed. [3H]D51-4700 was photo incorporated in several polypeptides but only the alpha1 subunit of the Ca2+ channel was photolabeled in a specific manner. Antibody mapping of the [3H]D51 4700-labeled alpha1 subunit with several anti-peptide antibodies revealed that the labeled site was located solely in a peptide fragment between Cys1461 and Lys1529. This region encompasses the labeled site of skeletal muscle, but contains several non-identical amino acid residues, which may participate in expressing different binding characteristics between the two muscle type Ca2+ channels. PMID- 9877172 TI - The artificial alpha1beta1-contact mutant hemoglobin, Hb Phe-35beta, shows only small functional abnormalities. AB - It was previously reported that Hb Philly with a mutation of Phe for Tyr at 35(C1)beta showed non-cooperative oxygen binding with a very high affinity and instability leading to hemolysis. Further, it lacked the 1H-NMR signal at 13.1 ppm from 2,2-dimethyl-2-silapentane-5-sulfonate in normal hemoglobin (Hb A), so that this signal was assigned to a hydrogen bond formed by Tyr-35(C1)beta. Surprisingly, our artificial mutant hemoglobin with the same mutation as Hb Philly showed slightly lowered oxygen affinity, almost normal cooperativity, the 1H-NMR signal at 13.1 ppm and no sign of instability. Our results indicate that the mutation reported for Hb Philly and the assignment of the 13.1 ppm signal need reexamination. PMID- 9877171 TI - Tumor-derived EMMPRIN (extracellular matrix metalloproteinase inducer) stimulates collagenase transcription through MAPK p38. AB - EMMPRIN (extracellular matrix metalloproteinase inducer) stimulates fibroblast metalloproteinases (MMP) 1, 2 and 3 (Kataoka et al. (1993) Cancer Res. 53, 3154 3158). Here we focus on MMP-1, showing that in lung tumors, MMP-1's cognate mRNA is strongly expressed in stromal fibroblasts adjacent to EMMPRIN-expressing tumor cells. In vitro, EMMPRIN upregulates MMP-1 mRNA expression in a concentration dependent manner, with a peak accumulation at 24 h. The response is genistein sensitive, suggesting it is dependent on tyrosine kinase activity. Analysis of tyrosine phosphorylation-dependent MAP kinases ERK 1/2, SAPK/JNK, and p38 showed that the activity of p38 but not that of the other 2 kinases was elevated in response to EMMPRIN. That p38 activity was required for EMMPRIN stimulation of MMP-1 was evident from results showing that the p38 inhibitor SB203580 blocked this response. This is the first available information regarding the mechanism by which tumor-associated molecules upregulate MMP synthesis in stromal fibroblasts. PMID- 9877173 TI - Changes in cytoplasmic ATP concentration parallels changes in ATP-regulated K+ channel activity in insulin-secreting cells. AB - Changes in cytoplasmic ATP concentration were monitored in intact insulin producing cells and correlated to changes in the activity of ATP-sensitive K+ channels (KATP channels). Luciferase was introduced into HIT M2.2 cells and whole pancreatic islets by transient expression of firefly (Photinus pyralis) luciferase cDNA. In transfected cells, extracellular addition of luciferin increased the luminescence signal to a maximum within 50-120 s. Addition of 1 microM of the mitochondrial uncoupler FCCP decreased the luminescence, an effect partly reversed upon withdrawal of the compound. High concentrations of glucose increased cytoplasmic free ATP concentration. Changes in the luminescence signal were accompanied by changes in activity of the ATP-sensitive K+-channel. Transfection per se did not deteriorate cell function, as verified by experiments showing similar changes in cytoplasmic free Ca2+-concentration, [Ca2+li, in both transfected and non-transfected cells. By measuring the cytoplasmic ATP concentration and KATP channel activity under similar experimental conditions, it was possible to establish, for the first time, a direct relationship between these two parameters. This indeed suggests that the cytoplasmic ATP concentration has a crucial role in the regulation of KATP channel activity under physiological conditions. PMID- 9877174 TI - Erythrosin 5'-isothiocyanate labels Cys549 as part of the low-affinity ATP binding site of Na+/K+-ATPase. AB - The high-affinity E1ATP site of Na+/K+-ATPase labeled with fluorescein 5' isothiocyanate and its E2ATP site labeled with erythrosin 5'-isothiocyanate (ErITC), as was shown recently [Linnertz et al. (1998) J. Biol. Chem. 273, 28813 28821], reside on separate and adjacent catalytic alpha subunits. This paper provides evidence that specific labeling of the E2ATP binding site with ErITC resulted in a modification of the Cys549 residue in the tryptic fragment with the sequence Val545-Leu-Gly-Phe-Cys549-His550. Hence, Cys549 is part of or close to the low-affinity E2ATP binding site of Na+/K+-ATPase. PMID- 9877176 TI - Uni-axial cyclic stretch induces c-src activation and translocation in human endothelial cells via SA channel activation. AB - The kinase activity of c-src increased and peaked at 15 min after an application of uni-axial cyclic stretch in HUVECs followed by a translocation of c-src to Triton-insoluble fraction. Suppression of c-src by an antisense S oligodeoxynucleotide inhibited the stretch-induced tyrosine phosphorylation and morphological changes. The stretch-induced increase in c-src activity was inhibited by FK506, a specific inhibitor for calcineurin, by Gd3+, a blocker for stretch activated channels, and by the extracellular Ca2+ depletion suggesting the involvement of SA channels. These results strongly suggest c-src plays an important role in the downstream of SA channel activation followed by the morphological changes. PMID- 9877175 TI - Positive feedback regulation of type I IFN genes by the IFN-inducible transcription factor IRF-7. AB - The interferon regulatory factor (IRF) family of transcription factors regulate the interferon (IFN) system, among which IRF-3 is involved in the virus-induced IFN-beta gene expression. Here we show that another member IRF-7 is critical for the IFN-alpha gene induction. Unlike the IRF-3 gene, the IRF-7 gene is induced by IFNs through activation of the ISGF3 transcription factor, and IRF-7 undergoes virus-induced nuclear translocation. In cells lacking p48, an essential component of IFN stimulated gene factor 3 (ISGF3), ectopic expression of IRF-7 but not IRF 3 can rescue the deficiency to induce IFN-alpha genes. These results indicate that IRF-7 is a key factor in the positive feedback regulation of IFN-alpha/beta production. PMID- 9877177 TI - Specific detections of the early process of the glycation reaction by fructose and glucose in diabetic rat lens. AB - The glycation reaction by fructose, as well as that by glucose, in control and diabetic rat lens was analyzed by using antibodies which specifically recognize adducts of lysine with fructose and with glucose. Levels of fructose adducts in diabetic rat lens were 2.5 times that of the control, and correlated with sorbitol levels. This was mainly due to enhanced glycation of beta- and gamma crystallins by fructose under diabetic conditions. These data suggest that glycation by fructose may also play a role in cataract formation under conditions of diabetes and aging. PMID- 9877178 TI - Localization of Rho GTPase in sea urchin eggs. AB - We isolated the urho1 (urchin rho in English or uni rho in Japanese) gene from the sea urchin cDNA library which encodes a Rho GTPase. Anti-URho1 antibodies specifically recognized a 22 kDa protein in the extracts of echinoderm eggs. URho1 was concentrated in the cortices from both unfertilized and fertilized eggs as judged by immunoblot analysis. URho1 may bind directly to the cell membrane but not be a component of the cortical layer. Immunofluorescence microscopy revealed that URho1 is localized to the cleavage furrow and the midbody during cytokinesis. PMID- 9877179 TI - A novel human RasGAP-like gene that maps within the prostate cancer susceptibility locus at chromosome 1q25. AB - We report the molecular cloning of a human cDNA that encodes a molecule having striking homology with Ras-specific GTPase-activating proteins (RasGAPs). Among previously described RasGAPs, the cDNA product is most closely related to Caenorhabditis elegans GAP-2, including a predicted coiled-coil structure near the carboxyl terminus. Expression of the cDNA in Saccharomyces cerevisiae defective in one of two RasGAPs, Ira2, complemented loss of the Ira2 function, indicating that the cDNA product functions as a RasGAP. The RasGAP-like gene is located on the human chromosome 1q25, the locus that appears to contain a hereditary prostate cancer susceptible gene, HPC1. PMID- 9877180 TI - Calmodulin dependence of NFkappaB activation. AB - The NFkappaB family of transcription factors is regulated by inhibitory IkappaB proteins. A diversity of stimuli leads to the phosphorylation and subsequent degradation of IkappaB, releasing NFkappaB to act on its target genes. Calmodulin (CaM) is a key regulator of numerous cellular processes and is the predominant intracellular receptor for Ca2+ signals. Here we report that several CaM antagonists inhibit the activation of NFkappaB, and that this is due to the prevention of inducible IkappaB phosphorylation. Our results suggest that CaM is involved in the phosphorylation of IkappaB, a finding that may help in elucidating the mechanism of this critical step of NFkappaB activation. PMID- 9877181 TI - A matrix metalloproteinase inhibitor which prevents fibroblast-mediated collagen lattice contraction. AB - Matrix metalloproteinases (MMPs) and the specific tissue inhibitors of metalloproteinases (TIMPs) are involved in tissue turnover in normal and pathological processes including wound healing. Marimastat, a potent inhibitor of MMPs, was used to investigate the role of MMPs in an in vitro wound contraction model, the dermal equivalent, in which fibroblasts are grown in a collagen matrix. Marimastat inhibited fibroblast-mediated lattice contraction and this inhibition was reversible upon removal of the inhibitor, indicating that MMPs play an important role in fibroblast-mediated collagen lattice contraction, modelling what may happen when granulation tissue contracts in a healing wound. PMID- 9877182 TI - Expression of a tyrosine phosphorylated, DNA binding Stat3beta dimer in bacteria. AB - The signal transducer and activator of transcription (STAT) proteins deliver signals from the cell membrane to the nucleus. An N-terminally truncated fragment of murine Stat3beta, Stat3betatc (127-722), was produced in bacteria. STAT proteins must be specifically phosphorylated at a single tyrosine residue for dimerization and DNA binding. Therefore, Stat3betatc was coexpressed with the catalytic domain of the Elk receptor tyrosine kinase. Stat3betatc was quantitatively phosphorylated by this kinase domain. Gel filtration chromatography revealed a Stat3betatc dimer. Y705 was identified as the major phosphorylated residue of Stat3betatc. This corresponds to the tyrosine residue which is phosphorylated by the Janus kinase in vivo. The phosphorylated Stat3betatc specifically bound to DNA binding sites. The described protocol allows the production of large amounts of activated protein for biochemical and pharmaceutical studies. PMID- 9877183 TI - Palmitoylation of the rat mu opioid receptor. AB - We examined whether the mu opioid receptor was palmitoylated and attempted to determine sites of palmitoylation. Following metabolic labeling with [3H]palmitic acid and immunoaffinity purification of the mu opioid receptor, SDS-PAGE and fluorography revealed a broad labeled band with Mr of approximately 80 kDa in CHO cells stably expressing the rat mu receptor, but not in CHO cells transfected with the vector alone, indicating that the mu receptor is palmitoylated. Activation of the receptor with morphine did not affect the extent of palmitoylation. Hydroxylamine or dithiothreitol treatment removed most of the radioactivity, demonstrating that [3H]palmitic acid is incorporated into Cys residue(s) via thioester bond(s). Surprisingly, mutations of the only two Cys residues in the C-terminal domain did not reduce [3H]palmitic acid incorporation significantly. Thus, unlike many G-protein coupled receptors, the palmitoylation site(s) of the rat mu opioid receptor do(es) not reside in the C-terminal domain. PMID- 9877184 TI - GSTP1-1 stereospecifically catalyzes glutathione conjugation of ethacrynic acid. AB - Using 1H NMR two diastereoisomers of the ethacrynic acid glutathione conjugate (EASG) as well as ethacrynic acid (EA) could be distinguished and quantified individually. Chemically prepared EASG consists of equal amounts of both diastereoisomers. GSTP1-1 stereospecifically catalyzes formation of one of the diastereoisomers (A). The GSTP1-1 mutant C47S and GSTA1-1 preferentially form the same diastereoisomer of EASG as GSTP1-1. Glutathione conjugation of EA by GSTA1-2 and GSTA2-2 is not stereoselective. When human melanoma cells, expressing GSTP1 1, were exposed to ethacrynic acid, diastereoisomer A was the principal conjugate formed, indicating that even at physiological pH the enzyme catalyzed reaction dominates over the chemical conjugation. PMID- 9877185 TI - The changing face of rheumatic fever in the 20th century. PMID- 9877186 TI - Antigenic and genomic homogeneity of successive Mycoplasma hominis isolates. AB - Sixty Mycoplasma hominis isolates were obtained from the cervices of pregnant women and from the ears or pharynges of their newborn babies. The isolates were examined by SDS-PAGE and pulsed-field gel electrophoresis. Antigenic and genomic profiles were obtained for 16 series with two or more successive isolates. Both analyses led to the conclusion that isolates from the same woman were identical or nearly identical, while isolates from different women exhibited a high degree of variation with respect to both genomic and antigenic profiles. PMID- 9877187 TI - An evaluation of intergenic rRNA gene sequence length polymorphism analysis for the identification of Legionella species. AB - There are currently more than 40 species of Legionella and the identification of most of these by standard methods is technically difficult. The aim of this study was to assess the suitability of a previously published PCR-based method of identifying Legionella spp. Intergenic 16S-23S rDNA spacer regions were amplified with primers complementary to conserved regions of the rRNA genes. Following electrophoretic separation of the products, data analyses were performed with the Taxotron software package. Computer-assisted analysis (with an empirically derived error tolerance of 3%) could differentiate only 26 of the 43 strains (representing 43 species), with the remaining 17 species clustering into four groups (group I, comprising 10 species; group II, three species; group III, two species and group IV, two species). Analysis of well-characterised 'non-type' strains of some Legionella spp. (e.g., from type culture collections) resulted in patterns distinct from the corresponding type strain in most cases. Furthermore, recent isolates (identified by conventional methods) were identified by this PCR method to the presumed correct species (or species group) in only a minority of cases. Well characterised strains and recent isolates of Legionella showed heterogeneity within many species. This intra-species variation severely limits the usefulness of the method for the identification of isolates. However, this property may be useful for epidemiological typing within such species. PMID- 9877188 TI - Demonstration that Australian Pasteurella multocida isolates from sporadic outbreaks of porcine pneumonia are non-toxigenic (toxA-) and display heterogeneous DNA restriction endonuclease profiles compared with toxigenic isolates from herds with progressive atrophic rhinitis. AB - Capsular types A and D of Pasteurella multocida cause economic losses in swine because of their association with progressive atrophic rhinitis (PAR) and enzootic pneumonia. There have been no studies comparing whole-cell DNA profiles of isolates associated with these two porcine respiratory diseases. Twenty-two isolates of P. multocida from diseased pigs in different geographic localities within Australia were characterised genotypically by restriction endonuclease analysis (REA) with the enzyme CfoI. Seven of 12 P. multocida isolates from nasal swabs from pigs in herds where PAR was either present or suspected displayed a capsular type D phenotype. These were shown to possess the toxA gene by polymerase chain reaction (PCR) and Southern hybridisation, and further substantiated by production of cytotoxin in vitro. The CfoI profile of one of these seven isolates, which was from the initial outbreak of PAR in Australia (in Western Australia, WA), was identical with profiles of all six other toxigenic isolates from sporadic episodes in New South Wales (NSW). The evidence suggests that the strain involved in the initial outbreak was responsible for the spread of PAR to the eastern states of Australia. Another 10 isolates, representing both capsular types A and D, were isolated exclusively from porcine lung lesions after sporadic outbreaks of enzootic pneumonia in NSW and WA. CfoI restriction endonuclease profiles of these isolates revealed considerable genomic heterogeneity. Furthermore, none of these possessed the toxA gene. This suggests that P. multocida strains with the toxA gene do not have a competitive survival advantage in the lower respiratory tract or that toxin production does not play a role in the pathology of pneumonic lesions, or both. REA with polyacrylamide gel electrophoresis and silver staining was found to be a practical and discriminatory tool for epidemiological tracing of P. multocida outbreaks associated with PAR or pneumonia in pigs. PMID- 9877189 TI - Flagellin gene variation between clinical and environmental isolates of Burkholderia pseudomallei contrasts with the invariance among clinical isolates. AB - The flagellin gene sequence from a clinical isolate of Burkholderia pseudomallei was used to design oligonucleotide primers for PCR/RFLP analysis of flagellin gene variation among clinical and environmental isolates of B. pseudomallei. Genes from four clinical and six environmental isolates were amplified and compared by RFLP. The clinical isolates were indistinguishable, but variation was detected among some of the environmental isolates. Sequence analysis of flagellin gene amplified products demonstrated high levels of conservation amongst the flagellin genes of clinical isolates (>99% similarity), compared to the variation observed between the clinical isolates and one of the environmental isolates (<90% similarity). Genomic comparisons with pulsed-field gel electrophoresis (PFGE) revealed differences between the relationships inferred by flagellin genotyping and PFGE, suggesting that a combination of molecular methods may be useful for the subtyping of B. pseudomallei strains. PMID- 9877190 TI - Identification of Helicobacter in gastric biopsies by PCR based on 16S rDNA sequences: a matter of little significance for the prediction of H. pylori associated gastritis? AB - The aim of the present study was to correlate molecular evidence of the presence of Helicobacter pylori in gastric biopsy samples, based on analysis of 16S rDNA, vacuolating toxin (vacA), urease A (ureA) and cagA genes, with the clinical, histological and serological findings in patients with H. pylori-associated gastritis. Fresh biopsy samples were collected from the gastric antrum and corpus of 22 asymptomatic volunteers with or without H. pylori-associated gastritis. Total DNA was extracted from the biopsy material and subjected to 16S rDNA PCR amplification, Southern blotting and 16S rDNA sequence analysis of the PCR products. The vacA, ureA and cagA genes were characterised by PCR amplification and Southern blot analysis. Based on partial 16S rDNA sequence analysis, DNA belonging to the genus Helicobacter was detected in gastric biopsy samples from 20 of 22 subjects, including seven of nine histologically and serologically normal controls. Six of 20 partial 16S rDNA sequences revealed variations within variable regions V3 and V4 that deviated from those of the H. pylori type strain ATCC 4350T and, therefore, possibly represented other species of Helicobacter. VacA genes identical with those of the type strain were found predominantly in the subjects with H. pylori gastritis, and all the patients except one were found to be cagA-positive. There was no evidence of false positive PCR reactions. In conclusion, the PCR-based molecular typing methods used here were apparently too sensitive when applied to the detection of H. pylori in human gastric tissues. The lack of quantitative analysis makes them inappropriate as clinical tools for the diagnosis of H. pylori-associated gastritis, despite the fact that they provide a qualitative and sensitive tool for the detection and characterisation of H. pylori in the gastrointestinal tract. PMID- 9877191 TI - The effect of culture conditions on the in-vitro adherence of methicillin resistant Staphylococcus aureus. AB - Methicillin-resistant isolates of Staphylococcus aureus (MRSA) were divided on the basis of their epidemiologic behaviour into two subgroups, epidemic MRSA (EMRSA) and sporadic MRSA (SMRSA) strains. An existing adherence assay was modified to determine differences in adherence properties between these two groups of MRSA, and the influence of culture conditions on the adherence of SMRSA and EMRSA strains to plastic, human collagen I (HuCol I) and pharyngeal carcinoma Detroit 562 cells (D562) was determined. In-vitro parameters, such as culture medium, growth temperature and growth phase of the bacterium, influenced the adherence of MRSA strains to plastic significantly. Even more pronounced differences in adherence due to changes in growth conditions and growth phase of the bacteria were found for the adherence of MRSA strains to HuCol I. Growth phase had a significant effect on the adherence of MRSA strains to the pharyngeal carcinoma cells D562. However, the study did not find conditions which made it possible to distinguish EMRSA from SMRSA strains. These data show that extrapolation of in-vitro data concerning adherence of MRSA strains to in-vivo conditions should be treated with caution. PMID- 9877192 TI - Expression of heat-shock proteins in Streptococcus pyogenes and their immunoreactivity with sera from patients with streptococcal diseases. AB - The heat-shock response of Streptococcus pyogenes following exposure to elevated growth temperatures, and the immunological reactivity of heat-shock proteins (HSPs) in streptococcal infections were studied. Two major proteins of 65 and 75 kDa were expressed when a S. pyogenes strain was shifted from 37 degrees C to heat-shock temperatures of 40, 42 and 45 degrees C. Such proteins are members of the GroEL and DnaK families recognised in a Western blot assay with polyclonal antibodies against Escherichia coli GroEL and E. coli DnaK, respectively. Two dimensional autoradiograms of polypeptides labelled at 37 or 42 degrees C showed an increased intensity of three spots at 42 degrees C. A monoclonal antibody (MAb) against HSP 63 of Bordetella pertussis also recognised the 65-kDa inducible protein, although MAbs against Mycobacterium tuberculosis HSP 65 failed to recognise this protein. Immunoblot analysis of sera from individuals with rheumatic fever or uncomplicated streptococcal diseases revealed seven major immunogenic protein bands, two of which also reacted with anti-E. coli GroEL and DnaK polyclonal antibodies. Furthermore, antibodies to the GroEL and DnaK proteins were also detected in sera from patients with either rheumatoid arthritis or systemic lupus erythematosus. These results demonstrated a heat shock response of S. pyogenes, and indicated the presence of an immune response against HSPs in streptococcal diseases. PMID- 9877193 TI - Role of group B streptococcal capsular polysaccharides in the induction of septic arthritis. AB - The ability of different serotypes of group B streptococci (GBS) to induce septic arthritis in mice was compared. Types II, III, IV, V, VI and VII GBS were investigated. A highly capsulate strain of type III GBS, COH1, and its mutants, COH1-11 (lacking capsular sialic acid) and COH1-13 (non-capsulate), obtained by transposon insertional mutagenesis, were used to assess the role of type-specific polysaccharide on the induction of arthritis. At an intravenous dose of 10(7) cfu/mouse, reference strains of types II, III, IV, VI and VII and type III strain COH1 induced arthritis with an incidence ranging from 70 to 90%. For type V and strain COH1-11, 10(8) cfu/mouse was required to obtain a 50% incidence of arthritis; lesions were not evident with strain COH1-13. The presence of the capsule played a major role in the induction of GBS septic arthritis. The presence and amount of sialic acid in capsular polysaccharide influenced the incidence of articular lesions. The bacterial dose affected the manifestations of arthritis; the less virulent strains of GBS also induced articular lesions when an adequate number of micro-organisms reached the joints. PMID- 9877194 TI - Mycobacterium avium infection of gut mucosa in mice associated with late inflammatory response and intestinal cell necrosis. AB - Mycobacterium avium is an intracellular pathogen that is associated with disseminated infection in acquired immunodeficiency syndrome (AIDS). Patients with AIDS appear to acquire M. avium mainly through the gastrointestinal tract. Previous studies have shown that healthy mice given M. avium orally develop disseminated infection after 2-4 weeks. The chief site of M. avium invasion of the intestinal mucosa is the terminal ileum. To learn more about the pathophysiology of M. avium infection of the intestinal mucosa, C57BL/6 bg+ bg+ mice were infected orally with M. avium strain 101 and groups of six mice were killed each week for 8 weeks. The terminal ileum was then prepared for histopathological studies and electron microscopy. A delayed inflammatory response was observed and influx of neutrophils in the Peyer's patches was the only abnormality seen at 1 week. A severe inflammatory response was seen from week 2 to week 5 and necrosis of intestinal villi was observed 6 weeks after infection. These results indicate that invasion and infection of the normal intestine by M. avium results in a severe inflammatory response with segmental necrosis of the intestinal mucosa. PMID- 9877195 TI - Detection of JC virus in two African cases of progressive multifocal leukoencephalopathy including identification of JCV type 3 in a Gambian AIDS patient. AB - Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating central nervous system (CNS) infection, affecting mainly oligodendrocytes, but also occasional astrocytes. In the USA, Europe and Asia, PML is caused by the human polyomavirus JC virus (JCV) and in autopsy series occurs in about 4-7% of AIDS patients. In Africa, the prevalence of PML in AIDS patients is uncertain and the causative agent is unknown. This study reports immunocytochemical and PCR confirmation of PML in the CNS of an AIDS patient dying in Uganda, East Africa (case 1). In a Gambian patient infected with HIV-2 who died 3 months after onset of AIDS/PML in Germany (case 2), it was possible to confirm the identity of the virus by DNA sequencing of the PCR amplified JCV product. This African genotype of the virus (type 3) showed an unusual re-arrangement of the regulatory region, and could be distinguished at several sites from East African and African American JCV strains described previously. This study has confirmed that PML is a complication of African AIDS as it is in Europe and the USA, and that JCV type 3 is pathogenic in African AIDS patients. Furthermore, the finding of an African genotype of JCV in a patient dying in Germany suggests that in this individual JCV represented a latent infection acquired in Africa. PMID- 9877196 TI - Laboratory and clinical comparison of preservation media and transport conditions for survival of Actinobacillus actinomycetemcomitans. AB - The capacity of clinical isolates and type strains of Actinobacillus actinomycetemcomitans to survive in a new transport medium (AaTM), phosphate buffered saline (PBS) and Ringer's solution (RS) was evaluated. The effects of exposure to air, transportation time and temperature on viability were also studied. In addition, the culture of A. actinomycetemcomitans from subgingival plaque of patients with different forms of periodontitis was quantified. The results following storage in AaTM, PBS and RS showed that A. actinomycetemcomitans survived better in AaTM than in PBS or RS when transportation times exceeded 20-22 h, and that survival was enhanced by storage at below 12 degrees C. Serotype b strains of A. actinomycetemcomitans were able to survive better than either serotype a or c. In the clinical study the optimal transportation conditions for subgingival plaque containing A. actinomycetemcomitans were AaTM at a temperature of 8 degrees C for 24 h under anaerobic conditions. These conditions resulted in a high survival and isolation rate for A. actinomycetemcomitans without inhibition of the other periodontopathic bacteria isolated from deep periodontal pockets. These findings have practical implications for future multicentre clinical trials in which the transportation of oral specimens over relatively long distances and at different ambient temperatures during various periods of the year are required. PMID- 9877197 TI - Incubation of superoxide dismutase with malondialdehyde and 4-hydroxy-2-nonenal forms new active isoforms and adducts. An evaluation of xenobiotics in fish. AB - The effects in fish (Sparus aurata) of dieldrin, previously reported to be an inducer of peroxisomal enzymes (Pedrajas et al., Comp. Biochem. Physiol. 115C (1996) 125-131), were compared with those of clofibrate. Although dieldrin provoked the more severe peroxisomal changes, both compounds induced oxidative stress as detected by the increased levels of microsomal thiobarbituric acid reactive substances; however the malondialdehyde (MDA) content, determined after HPLC separation of the MDA-TBA complex, was not significantly altered. These results suggest that, besides MDA, other aldehydes were formed in xenobiotic injected fish, leading us to assess the oxidative effects of such xenobiotics by following changes in superoxide dismutase (SOD) pattern. New active SOD isoforms were detected by isoelectrofocusing in the light mitochondrial (LMF) and cytosolic (CF) fractions. Most of the new SOD bands could be reproduced in vitro by incubation of fish liver cell-free extracts with MDA. To clarify the effects of aldehydes, Cu,Zn- and Mn-SOD isoforms were purified and amino acid analysis was carried out. The new bands found in LMF and CF fractions were reproduced in vitro after incubation of pure SODs with MDA and 4-hydroxy-2-nonenal (HNE), the new SOD bands formed being coincident with the loss of Lys or His residues. Lysine residues were preferentially derivatized after treatment of Cu,Zn-SOD with MDA, but in Mn-SOD the lysine residues were modified only after treatment with MDA, while the histidine residues were modified only by HNE. No change of SOD activity was detected after MDA or HNE exposure, although at the higher aldehyde concentrations used protein aggregates were formed. Therefore, the appearance of new active SOD bands, after isoelectrofocusing separation, can be proposed as a biomarker of oxidative stress. PMID- 9877198 TI - Comparison of the mode of action of a dinuclear platinum complex containing a pyridine derivative with its monomeric analog. AB - The DNA binding and interstrand cross-linking properties of the dinuclear platinum complex [?cis-Pt(NH3)2Cl?2bpsu](NO3)2 (bpsu is 4,4'-dipyridyl sulfide) (II) and the mononuclear complex [cis-Pt(NH3)2Cl(4-methylpyridine)]NO3 (I) were compared with those of [?cis-Pt(NH3)2Cl?2H2N(CH2)4NH2](NO3)2 (III) in order to understand the mode of action of complexes I and II. Both compound I and compound II caused significantly different changes of conformation in poly(dG-dC) x poly(dG-dC) than compound III did. Studies of DNA binding, interstrand cross linking and fluorescence assay suggest that compound I monofunctionally binds to DNA and compound II bifunctionally binds to DNA, that the dinuclear platinum complex II more efficiently interacts with DNA compared to its monomeric analog, and that platinum I and II complexes both interact with DNA in a non intercalative mode. All the results indicate that the mode of action of the dinuclear complex II is different from that of the mononuclear complex I. PMID- 9877200 TI - Mechanism of mitochondrial dysfunction and cytotoxicity induced by tropolones in isolated rat hepatocytes. AB - The mechanism of mitochondrial dysfunction and toxicity induced by the tropolones, beta-thujaplicin (4-isopropyl tropolone), tropolone and tropone, has been studied in freshly isolated rat hepatocytes. Incubation of hepatocytes with beta-thujaplicin (1-4 mM) elicited a concentration and time-dependent cell killing. The toxicity was accompanied by losses of cellular ATP, total adenine nucleotides and glutathione, independently of lipid peroxidation and protein thiol oxidation. The beta-thujaplicin-induced cytotoxicity was enhanced by the pretreatment of hepatocyte suspensions with EDTA (4 mM), a hydrophilic chelator, or by incubation in Ca2+ and Mg2+-deficient Krebs-Henseleit buffer. The partition coefficient of beta-thujaplicin, which formed complex with the divalent cations in Krebs-Henseleit buffer, in n-octanol/buffer was increased either in the presence of EDTA or absence of divalent cations. Comparison of toxic effects based on cell viability and adenine nucleotide levels showed that beta thujaplicin was more toxic than tropolone or tropone in Krebs-Henseleit buffer containing EDTA (4 mM). The addition of beta-thujaplicin to isolated hepatic mitochondria reduced state 3 respiration with NAD+-linked substrate (pyruvate plus malate) and/or with an FAD-linked substrate (succinate plus rotenone), whereas state 3 respiration of ascorbate plus tetramethyl-p-phenylenediamine (cytochrome oxidase-linked respiration) was not significantly affected by beta thujaplicin. Further, the addition of these tropolones caused a concentration dependent increase in the rate of state 4 oxygen consumption, indicating an uncoupling effect. These results indicate that beta-thujaplicin- and tropolone induced cytotoxicity are associated with an acute ATP depletion via mitochondrial dysfunction related to oxidative phosphorylation and that the induction of cytotoxicity is affected by EDTA or divalent cations. PMID- 9877199 TI - Glutathione conjugation of perchloroethene in subcellular fractions from rodent and human liver and kidney. AB - Perchloroethene (Per) is a widely used industrial solvent and common environmental contaminant. In rats, long-term inhalation of Per is known to cause a small increase in the incidence of renal tubule cell tumors in males only; renal toxicity is seen in female rats and in both sexes of mice after prolonged Per exposure. The renal toxicity of Per is likely mediated by a glutathione dependent bioactivation reaction. Glutathione S-transferase mediated formation of S-(1,2,2-trichlorovinyl)glutathione is the first step in a sequence of reactions finally resulting in the formation of reactive intermediates in the kidney. In this study, we compared the enzymatic rates of formation of S-(1,2,2 trichlorovinyl)glutathione in liver and kidney subcellular fractions from rats, mice, and from both sexes of humans (n = 11). In microsomal fractions from the liver and kidney of all three species, enzymatic formation of S-(1,2,2 trichlorovinyl)glutathione from Per could not be observed. S-(1,2,2 Trichlorovinyl)glutathione formation (the structure was confirmed by electrospray mass spectrometry) was observed in liver cytosol from both male and female rats and mice. However, the rates of S-(1,2,2-trichlorovinyl)glutathione formation in liver cytosol from male rats (84.5+/-12 pmol/mg per min) were approximately four times higher than from female rats (19.5+/-8 pmol/mg per min) and from both sexes of mice (27.9+/-6 and 26.0+/-4 pmol/mg per min). Low rates of S-(1,2,2 trichlorovinyl)glutathione formation were also seen in kidney cytosol from mice (12+/-6 pmol/mg per min), but not from rats. In human liver subcellular fractions, enzymatic formation of S-(1,2,2-trichlorovinyl)glutathione could not be detected. The human liver cytosolic fractions, however, exhibited glutathione S-transferase activity (as determined using 1-chloro-2,4-dinitrobenzene and hexachlorobutadiene as marker substrates) in the same order of magnitude as rat and mouse liver cytosol. In contrast to other marker activities for glutathione S transferases, the ability of all human liver cytosol samples to catalyze the glutathione conjugation of 1,2-dichloro-4-nitrobenzene was three orders of magnitude lower compared to rat and mouse liver cytosol. 1,2-Dichloro-4 nitrobenzene conjugation was also four times higher in liver cytosol from male rats compared to female rats. The results suggest that the ability of the human liver to catalyze the formation of S-(1,2,2-trichlorovinyl)glutathione from Per is at least two orders of magnitude lower than that of rat liver, and that sex specific differences in the extent of hepatic conjugation of Per with glutathione, which may be relevant for nephrotoxicity, occur in rats. PMID- 9877201 TI - Cell proliferation induced by 8-oxoguanosine and 8-methylguanosine, two adducts produced by free radical attack on ribonucleosides and RNA. AB - The ability of C8-substituted guanine (Gua) ribonucleosides to induce B cell proliferation has been well documented in the literature. These compounds are analogues of adducts formed from free radical attack on ribonucleosides and RNA. Here we examined the proliferative properties of two of these radical adducts, 8 methylguanosine (8-MeG) and 8-oxo-7,8-dihydroguanosine (8-OxoG) and compared them with those of the well studied B cell mitogen, 8-bromoguanosine (8-BrG). 8-MeG and 8-OxoG were synthesized in the considerable yields of 28, and 55%, respectively, and were characterized by UV, NMR and CG-MS. Their effects upon [3H]thymidine uptake into DNA by Swiss mouse splenocytes, mouse embryo 3T3 fibroblasts (A31) and mouse B16F10 melanoma were examined. Both guanosine (G) radical adducts were shown to increase [3H]thymidine uptake by mouse splenocytes but displayed selectivity in respect to continuous cell lines. 8-MeG acted upon 3T3 fibroblasts whereas 8-OxoG acted upon B16F10 melanoma. The non-physiological analogue 8-BrG acted upon all tested cells. Parallel experiments of cell counting, cytotoxicity,and cell sorting indicated that DNA synthesis induced by the C8-substituted G's reflected cell growth. It is proposed that the compounds act intracellularly because their proliferative effects were blocked in the presence of a nucleoside transport inhibitor but were not inhibited by an antagonist of the A2 purine receptor. The present results, taken together with data from the literature, suggest that in the case of 3T3 fibroblasts and mouse splenocytes the proliferative effects of the compounds are not dependent on metabolism through purine salvage pathways. In the case of melanoma, however, the compounds are likely to become part of the purine nucleoside pool. The demonstration that adducts produced by free radical attack on ribonucleosides and RNA are able to induce cell proliferation opens new perspectives for the understanding of free radical mediated carcinogenesis. PMID- 9877202 TI - Elevation of ubiquinone content by peroxisomal inducers in rat liver during aging. AB - The effect of di(2-ethylhexyl)phthalate (DEHP) on the induction of peroxisomes and the content of ubiquinone in the liver was studied in rats between 25 and 496 days of age. During this period, peroxisomal beta-oxidation of fatty acids was greatly decreased but it could be induced many-fold in all ages. The ubiquinone (UQ) content was increased upon induction 6-fold in the first weeks of life, but the extent of this elevation continuously narrowed and no induction could be observed in the oldest animals, even after prolonged treatment with the plasticizer. In contrast, the treatment decreased the amount of liver cholesterol in all age groups. Treatment with this peroxisomal inducer increased the biosynthesis of UQ while the breakdown rate was found to be unaffected, as the half-life of this lipid was 103 and 106 h in control and treated rats, respectively. These results indicate that treatment with peroxisomal inducers increases the liver UQ content by increasing the rate of biosynthesis and that this effect is not apparent in aged rats. PMID- 9877204 TI - Metabolic effects and distribution space of flufenamic acid in the isolated perfused rat liver. AB - The following aspects were investigated in the present work: (a) the action of flufenamic acid on hepatic metabolism (oxygen uptake, glycolysis, gluconeogenesis, uricogenesis and glycogenolysis), (b) the action of flufenamic acid on the cellular adenine nucleotide levels, and (c) the transport and distribution space of flufenamic acid in the liver parenchyma. The experimental system was the isolated perfused rat liver. Perfusion was accomplished in an open, non-recirculating system. The perfusion fluid was Krebs/Henseleit bicarbonate buffer (pH 7.4), saturated with a mixture of oxygen and carbon dioxide (95:5) by means of a membrane oxygenator and heated to 37 degrees C. The distribution space of flufenamic acid was measured by means of the multiple indicator dilution technique with constant infusion (step input) of [3H]water plus flufenamic acid. The results of the present work indicate that the metabolic effects of flufenamic acid are the consequence of an uncoupling of oxidative phosphorylation, a conclusion based on the following observations: (a) flufenamic acid increased oxygen uptake, a common property of all uncouplers; (b) the drug also increased glycolysis and glycogenolysis in livers from fed rats (these are expected compensatory phenomena for the decreased mitochondrial ATP formation); (c) flufenamic acid inhibited glucose production from fructose, an energy dependent process; (d) the cellular ATP levels were decreased by flufenamic acid whereas the AMP levels were increased; and (e) the total adenine nucleotide content was decreased by flufenamic acid and uric acid production was stimulated. Indicator-dilution experiments with flufenamic acid revealed that this substance undergoes flow-limited distribution in the liver and that its apparent distribution space greatly exceeds the aqueous space of the liver. Flufenamic acid changed its behaviour when the portal concentration was increased from 25 to 50 microM. At 25 microM the initial upslope of the outflow profile clearly preceded that of all other concentrations. From the trend of the curves obtained with 50, 100 and 250 microM, one would expect an initial upslope situated at the right of the 50-microM curve. Furthermore, the time of appearance of flufenamic acid in the outflowing perfusate was practically the same irrespective of the portal concentration. For theoretical reasons one would expect progressively longer appearance times when the portal concentration was decreased. It is possible that the amount of flufenamic acid bound to the cell membranes during the early stages of the infusion produced changes that enabled these structures to bind a larger quantity of the drug than originally possible. PMID- 9877203 TI - Involvement of capsaicin-sensitive nerves in paraquat-induced mortality. AB - Paraquat (PQ), a broad spectrum herbicide, produces severe lung inflammation and necrosis resulting in pulmonary fibrosis and respiratory failure. Tachykinins are peptides released by sensory C fibers and have the ability of influencing respiratory functions and cellular proliferation. To examine whether the damage caused by PQ involves tachykinins, rats were depleted in their content of tachykinins by systemic treatment with capsaicin prior to PQ exposure. The animal subjected to this treatment showed a 3-fold higher viability compared to those treated with PQ alone (75 vs 27%). Depletion of reduced glutathione (GSH) is associated with oxidative stress produced by reactive oxygen intermediates during PQ metabolism. This is considered to be critical in the pathogenesis of lung damage by PQ. PQ treatment induced a significant depletion of GSH during the first days and a similar effect was also observed in the group of capsaicin pretreated rats. Four weeks after PQ treatment the levels of GSH were similar to controls in rat pretreated or not with capsaicin plus PQ. This may indicate that the reduced levels of GSH may be associated to the toxicity observed in the acute phase, but not of importance in the final PQ-induced mortality. Neutral endopeptidase (NEP) is an enzyme considered to be critical in controlling the levels of tachykinins. Exposure of crude membrane preparations of rat lung to PQ resulted in a dose-dependent inhibition of NEP activity. Since NEP inactivation may occur in lung following a PQ exposure in vivo, the results indicate that during PQ intoxication a more sustained activity of tachykinins may be present, producing effects such as cell proliferation, fluid extravasation and bronchoconstriction. In conclusion, this finding supports the hypothesis that neuropeptides released from capsaicin-sensitive nerves could be involved in the modulation of PQ-induced lung damage. PMID- 9877205 TI - Interindividual variability in P450-dependent generation of neoantigens in halothane hepatitis. AB - Halothane hepatitis occurs because susceptible patients mount immune responses to trifluoroacetylated protein antigens, formed following cytochrome P450-mediated bioactivation of halothane to trifluoroacetyl chloride. In the present study, an in vitro approach has been used to investigate the cytochrome P450 isozyme(s) which catalyze neoantigen formation and to explore the protective role of non protein thiols (cysteine and reduced glutathione). Significant levels of trifluoroacetyl protein antigens were generated when human liver microsomes, and also microsomes from livers of rats pre-treated with isoniazid, phenobarbital or beta-naphtoflavone, were incubated with halothane plus a nicotinamide adenine dinucleotidephosphate (NADPH) generating system. Immunoblotting studies revealed that the major trifluoroacetyl antigens expressed in vitro exhibited molecular masses of 50-55 kDa and included 60 and 80 kDa neoantigens recognized by antibodies from patients with halothane hepatitis. Much lower concentrations of halothane were required to produce maximal antigen generation in isoniazid induced rat microsomes, as compared with phenobarbital or isosafrole-induced microsomes (0.5 vs 12.5 microl/ml). In isoniazid-induced microsomes, antigen generation was inhibited > 90% by the nucleophiles cysteine and glutathione and by the CYP2E1-selective inhibitors diallylsulfide and p-nitrophenol, but was unaffected by inhibitors of other P450 isozymes (furafylline, sulfaphenazole or triacetyloleandomycin). Neoantigen formation in six human liver microsomal preparations was inhibited in the presence of diallylsulfide, but not by furafylline, sulfaphenazole or triacetyloleandomycin, and exhibited marked variability which correlated with CYP2E1 levels. These results suggest that the balance between metabolic bioactivation by CYP2E1 and detoxication of reactive metabolites by cellular nucleophiles could be an important metabolic risk factor in halothane hepatitis. PMID- 9877206 TI - Alterations of small-molecular-weight antioxidants in the blood of smokers. AB - Plasma alpha-tocopherol, ascorbate, retinol, uric acid, and lipid peroxides were investigated in 39 male smokers and 64 male non-smokers. The average level of plasma alpha-tocopherol of 35-45-year-old smokers (1.74+/-0.49 microg/mg total lipid) was significantly lower than that of age-matched non-smokers (2.55+/-0.88 microg/mg total lipid, P = 0.032). Similarly, the plasma alpha-tocopherol of smokers aged above 45 (1.66+/-0.29 microg/mg total lipid) was lower than that of the age-matched non-smokers (2.38+/-1.26 microg/mg total lipid, P = 0.014). However, no difference in plasma level of alpha-tocopherol was found between smokers and non-smokers below the age of 35. The average concentration of ascorbate in plasma was significantly decreased only in those smokers older than 45 (0.33+/-0.16 mg/dl vs non-smokers 0.53+/-0.19 mg/dl, P = 0.003). The average level of lipid peroxides (measured as malondialdehyde,MDA) in the plasma of smokers (2.77+/-0.51 nmol/ml) was higher than that of non-smokers (2.35+/-0.21 nmol/ml) aged above 45 (P = 0.034). No differences in the plasma levels of uric acid and retinol were noted between smokers and non-smokers in all age groups. Using partial correlation analysis under age control, we found that the plasma level of alpha-tocopherol was negatively correlated with the plasma level of MDA (r = -0.523, P = 0.038). In contrast, the plasma level of ascorbate was only weakly correlated with the plasma level of MDA (r = -0.341, P = 0.094). Moreover, we found a negative correlation between the plasma level of alpha-tocopherol and smoking index (r = -0.414, P = 0.006) under age control, but there was no correlation between plasma level of ascorbate and smoking index (r=0.221, P = 0.193). These results indicate that adequate levels of alpha-tocopherol and ascorbate may protect the plasma from oxidative damage elicited by smoking mediated reactive oxygen species (ROS) and free radicals in young smokers. However, the antioxidant activities of alpha-tocopherol and ascorbate may be overwhelmed by the long-standing oxidative stress elicited by cigarette smoking in elderly subjects. PMID- 9877207 TI - The oestrogen receptor regulates NFkappaB and AP-1 activity in a cell-specific manner. AB - Oestrogens regulate the expression of genes both positively and negatively in a range of cell types. These effects are mediated via the oestrogen receptor (ER) and involve direct interactions between the ER and DNA response elements, as well as interactions between the ER and other nuclear proteins. We have examined the potential of the ERalpha to regulate the expression of reporter genes under the control of oestrogen response elements (EREs), NFkappaB response elements (NREs) or AP-1/TPA response elements (TREs) in HeLa cells and in human embryonic kidney (HEK-293) cells. Transiently transfected ERalpha was able to activate expression of beta-galactosidase under the control of EREs in an oestradiol (E2)-dependent manner in both HeLa and HEK-293 cells. The ERalpha was able to repress by 80% the TNF-mediated expression of beta-galactosidase under the control of NREs in an E2 dependent manner in HeLa cells but not in HEK-293 cells. ERalpha/E2 also induced a two-fold potentiation of TPA-mediated expression of beta-galactosidase under the control of TREs in HeLa cells but not in HEK-293 cells. These results suggest that the ERalpha is capable of regulating gene expression in a cell-specific manner. We further investigated the mechanisms by which the ERalpha regulates gene expression in these systems by co-expressing the ERalpha and the reporter gene constructs with known cofactors of the ERalpha. We have shown that expression of steroid receptor coactivator-1 alpha (SRC-1alpha) and receptor interacting protein-140 (RIP-140) have no effect on the capacity of the ERalpha to modulate NFkappaB reporter gene activity in HeLa cells. Furthermore, the expression of SRC-1alpha or RIP-140 does not enable the ERalpha to repress NFkappaB or to potentiate an AP-1 response in HEK-293 cells. This suggests that factors other than SRC-1alpha or RIP-140 are responsible for the cell-specific effects seen with ERalpha. PMID- 9877208 TI - cAMP activates transcription of the human glucocorticoid receptor gene promoter. AB - Glucocorticoids and cAMP regulate, either in a synergistic or additive fashion, the transcription of multiple genes, although some antagonistic effects of dexamethasone on cAMP-activated transcription have been described. The increased glucocorticoid receptor (GR) mediated response of some cell types, as a result of augmented cAMP, has been considered to be mainly due to an increased stability of GR mRNA, although other plausible explanations should not be ruled out. We studied the possibility that GR transcription itself could be affected by cAMP levels. HeLa cells were transfected with human GR (hGR) promoter constructs and their transcriptional activity determined after inducing a cAMP increase with forskolin. We found that forskolin almost doubled the transcriptional activity of the promoter construct spanning -2995 to +38 of the hGR, whereas no significant variations were observed with shorter chimeras containing sequences downstream 979. Shift mobility showed binding of CREB in vitro to a putative cAMP responsive element located at -1000, suggesting that hGR may be upregulated by cAMP at the transcriptional level, thus adding a new mechanism ascribable to this second messenger, which in conjunction with the cAMP-induced GR mRNA increased stability, would lead to a more precise control of the amount of GR protein within the cell. PMID- 9877210 TI - Involvement of glucocorticoid receptor in the induction of differentiation by ginsenosides in F9 teratocarcinoma cells. AB - We have previously reported that ginsenosides Rh1 and Rh2 induced the differentiation of F9 teratocarcinoma stem cells [Lee, Y. N., Lee, H. Y., Chung, H. Y., Kim, S. I., Lee, S. K., Park, B. C. and Kim, K. W., In vitro induction of differentiation by ginsenosides in F9 teratocarcinoma cells. Eur. J. Cancer 1996, 32, 1420-1428.]. Since the chemical structure of Rh1 and Rh2 is very similar to that of dexamethasone, a synthetic glucocorticoid, we investigated whether Rh1 and Rh2 act through the glucocorticoid receptor (GR). Immunocytochemistry showed that Rh1 or Rh2 increased the nuclear translocation of GR in the same manner of dexamethasone. In the gel shift assay, glucocorticoid response element (GRE) binding protein in F9 cells was increased by Rh1 or Rh2. To confirm whether the increased binding protein is GR, we performed the competition assay with unlabeled GRE as a specific competitor. Moreover, supershift assay with the GR antibody showed that the binding proteins are GR. In addition, to confirm the Rh1 or Rh2-induced transactivation of GRE promoter, we cotransfected GR expression vector and GRE-luciferase vector. In the luciferase assay, Rh1 or Rh2 potently induced luciferase activity and this induction was blocked by RU486, a potent GR antagonist. Taken together, we suggest that ginsenosides Rh1 and Rh2 may induce the differentiation of F9 cells by stimulating the nuclear translocation of GR. PMID- 9877209 TI - 4-iodotamoxifen aziridine, a new affinity labeling agent for the rapid detection of estrogen receptor isoforms. AB - We describe the simple and fast preparation of a new radioiodinated probe for the detection of the estrogen receptor (ER) and its isoforms. Iodotamoxifen aziridine was labeled with iodine 125 ([125I]TAZ) in position 4 of the alpha aromatic ring. The yield was high (>75%), the label was stable and the specific activity was near optimal (1900-2170 Ci/mmol). The apparent relative binding affinity of the probe to a recombinant human ER (hER) was high (RBA = 35 vs estradiol = 100). Electrophoretic studies (SDS-PAGE) with this hER indicated the high potency of [125I]TAZ at very low concentration (<1 nM) to reveal ER bands after a short exposure time (1-4 days). Competition between this probe and various compounds as well as chemical treatments of the ER with SH-reactive chemicals, demonstrated the labeling specificity. Analysis of cytosols from a panel of cell lines and various rat reproductive organs displayed characteristic ER bands (67, 50 and 37 kDa) suppressed by unlabeled E2. Detection in nonreproductive organs of 43 kDa E2 nondisplaceable peptide raised the question upon the presence of altered and/or variant ERs in many tissues. Data concerning human breast cancer cytosols were in complete accordance with those established with [3H]TAZ: high ER polymorphism in most ER-positive samples and peculiar forms (mainly 43 kDa) in ER-negative samples. Hence, [125I]TAZ appears especially useful for the detection of altered ER or related peptides in breast cancers. PMID- 9877211 TI - Later onset of apoptosis in the bulbourethral glands after castration compared to that in the seminal vesicles. AB - Androgens affect many different target organs within the male reproductive tract to stimulate their development and secretory cytodifferentiation, and to maintain structure and function in adulthood. Castration causes regression of these organs via apoptosis. However, not all organs of the reproductive tract are equally sensitive to androgen withdrawal. The effects of castration on the mouse seminal vesicles (SVs) and bulbourethral glands (BUGs) were compared in terms of protein and DNA contents, epithelial apoptosis, and proliferative response of epithelial cells to androgen. Castration induced similar, marked decreases in protein contents in the SV and BUG by 2 days after castration which reached a minimum at 16 days post castration. Both organs underwent a decrease in DNA content, but the kinetics of this decline differed. In the SV, DNA content was significantly decreased by 4 days whereas in the BUG this did not occur until 16 days post castration. By day 16 both organs had regressed to roughly the same degree. The apoptotic index in the epithelium reflected this difference in timing as well. Apoptotic index of the SV epithelium was highest on day 3 after castration and declined thereafter. On the other hand, the apoptotic index in the BUG didn't begin to increase until 7 days after castration and became maximal on day 12. Daily injections of testosterone propionate (TP) from day 8, 16, or 30 after castration all increased epithelial labelling index in the SVs to a similar degree. However, the TP-induced increase in the epithelial labelling index in the BUG beginning on day 8 after castration was considerably less than that in BUGs receiving TP treatment from day 16 or 30 after castration. Thus, the proliferative response of the epithelium depended upon prior apoptosis in the gland, with the timing being delayed in the BUG as compared with the SV. The present results indicate that castration induces epithelial apoptosis and reduction in glandular DNA content considerably later in the BUG than in the SV though reduction in protein content in the BUG fell simultaneously with that in the SV. PMID- 9877212 TI - Pharmacokinetics and pharmacodynamics of a novel estrogen delta8-estrone in postmenopausal women and men. AB - Recently a tenth equine estrogen, identified as the sulfate ester of delta8 estrone has been reported to be present in Premarin (a conjugated equine estrogen preparation), and because of its unique ring B unsaturated structure (conjugated double bond in the B ring), we have, in the present study, determined its pharmacokinetics in postmenopausal women and men, its interaction with uterine estrogen receptors and its uterotropic activity. After the administration of [14C]delta8-estrone, blood was drawn at various time intervals, and the plasma fractionated into the unconjugated sulfate and glucuronide fractions. The disappearance of radioactivity as delta8-estrone from plasma can be described as a function of two exponentials. The half-lives of the first and second components were 5+/-0.2 and 40.4 min, respectively. The mean metabolic clearance rate calculated (MCR), was 1711+/-252 l/d m2. From the unconjugated fraction, delta8 17beta-estradiol was also isolated and identified. From the sulfate conjugated fraction, delta8-estrone sulfate and delta8-17beta-estradiol sulfate were isolated in almost equal amounts. No other metabolites of delta8-estrone was detectable in the plasma. Both delta8-estrone and delta8-17beta-estradiol bind with human endometrial and rat uterine estrogen receptors with high affinity. The binding affinities of delta8-17beta-estradiol for human endometrial and rat uterine cytoplasmic receptors were 4 and 25 times higher than those of the parent estrogen delta8-estrone, respectively. Administration of delta8-estrone and delta8-17beta-estradiol (2 microg/100 g body weight) to immature rats significantly (P< 0.05) increased the uterine weight compared to the controls. These data demonstrate that delta8-estrone has estrogenic activity, and that it is further metabolized in man to a single more potent estrogen, delta8-17beta estradiol. The extent of this activation by 17beta-reduction appears to be greater than that observed with other estrogens. Both estrogens circulate as sulfate conjugates and are very slowly eliminated from the circulation. These data further suggest that delta8-estrone and its major metabolite delta8-17beta estradiol can contribute to the overall in vivo biological effects of Premarin. PMID- 9877213 TI - Androgen-induced vitellogenin gene expression in primary cultures of rainbow trout hepatocytes. AB - The hormone induction of vitellogenin (Vg) gene expression was investigated in primary cultures of immature male rainbow trout hepatocytes. Vg cDNA was cloned from the liver of a female rainbow trout injected with estradiol-17beta (E2). A Vg mRNA of about 6.6 kb was detected in the hepatocytes after stimulation with 2 x 10(-9) M E2 for 24 h. The intensity of this signal increased with E2 concentration. Several other hormones including progesterone and androgens (testosterone, androsterone, methyltestosterone) also induced Vg mRNA not only at high (2 x 10(-5) M) but also at low (2 x 10(-9) M), physiologically relevant steroid concentrations. Only cortisol failed to do so at any tested concentration but, surprisingly, like progesterone, it induced an additive effect in the presence of E2. On the other hand, tamoxifen drastically reduced the expression of Vg gene whether stimulated by E2 or any other tested steroid. In conclusion, our studies have highlighted the unusual specificity of Vg mRNA induction in fish. PMID- 9877214 TI - Determination by modified RT-PCR of transcript amounts from genes involved in sex steroid synthesis in chicken organs including brain. AB - Sex steroid hormones in mammals have shown to be synthesized not only in gonads but also in non-steroidogenic organs such as the brain. Steroid hormones in the brain were indicated to be involved in sex behavior and brain differentiation. In avian species, an experimental injection of androgen into the brain suggested the existence of a steroidogenic pathway. However, no studies have demonstrated the expression of genes involved in such a steroidogenic pathway in the brain, or in other non-steroidogenic organs of birds such as the liver and kidney. In this study, we have modified the RT-PCR procedure to analyze the expression of the steroidogenic genes, P-450scc, 3beta-HSD, P-450c17, 17beta-HSD and P-450arom in non-steroidogenic organs of chicken including the brain. The RT-PCR has demonstrated the presence of mRNAs from genes in non-steroidogenic as well as steroidogenic organs of chicken. The amounts of mRNAs from these genes (except for P-450c17) among the non-steroidogenic organs were found to be greatest in the brain. PMID- 9877216 TI - A time-resolved fluoroimmunoassay for cortisol in unextracted bovine plasma or serum with optimized procedures to eliminate steroid binding protein interference and to minimize non-specific streptavidin-europium binding. AB - A time-resolved fluoroimmunoassay (TR-FIA) for human salivary cortisol was adapted for the measurement of cortisol in unextracted bovine blood plasma and serum. It has been demonstrated that the binding of cortisol binding plasma proteins (CBPP) to the cortisol-biotin primary probe cannot be eliminated by means of cortisol releasing agents. Complete inactivation of CBPP was achieved by heating water diluted samples for 30 min at 80 degrees C. The high non-specific binding (NSB) of the streptavidin-europium secondary probe, encountered during preliminary experiments, was shown to be caused by an interaction with bovine serum albumin (BSA) and could be reduced partly by the addition of heparin. It was also shown that the ability to bind streptavidin-europium nonspecifically is not a general property of proteins since bovine gamma-globulin and gelatin lack this behaviour. The advantage of a highly reduced NSB, resulting from the use of a BSA free assay buffer, is not limited to this particular assay but is also beneficial for other procedures based on specific measurement of streptavidin europium fluorescence. The detection limit for a 20 microl sample was 0.5 ng/ml. The intra-assay coefficients of variation for control samples with cortisol concentrations of 71.1, 39.2 and 10.3 ng/ml were 8.2, 7.9 and 11.3% (n = 16). The corresponding inter-assay coefficients of variation were 7.3, 9.0 and 11.2% (n = 73). Correlation with a commercially available radioimmunoassay, preceded by diethylether extraction of the sample, was 0.97 (n = 88). PMID- 9877215 TI - Corticotropin-inhibiting peptide enhances aldosterone secretion by dispersed rat zona glomerulosa cells. AB - Corticotropin-inhibiting peptide (CIP), the 7-38 fragment of human ACTH(1-39), is known to act as an antagonist of ACTH receptors. Accordingly, CIP has been found to inhibit ACTH-stimulated glucocorticoid secretion of dispersed rat adrenocortical cells, without per se affecting the basal production. We confirmed these findings, but unexpectedly observed that CIP concentration-dependently raised basal aldosterone secretion from fresh suspensions of rat zona glomerulosa (ZG) cells, maximal effective concentration being 10(-6) M. CIP (10(-6) M) partially reversed the ZG-cell response to ACTH, but not to the Ca2+-dependent agonists angiotensin-II (ANG-II) and K+. The aspecific ANG-II-receptor antagonist saralasin (10(-6) M) blocked the aldosterone response of ZG cells to 10(-6) M CIP, and in the presence of the Ca2+-channel blocker verapamil CIP was ineffective. Collectively, these findings suggest that CIP enhances aldosterone secretion of rat ZG through a mechanism involving the activation of ANG-II receptors and the consequent rise in the cytosolic Ca2+ concentration. They also stress that this side-effect of CIP must be taken into account in interpreting the results of investigations on the adrenal cortex, where CIP has been employed as an ACTH-receptor antagonist. PMID- 9877217 TI - Progesterone 6-hydroxylation is catalysed by cytochrome P-450 in the moderate thermophile Bacillus thermoglucosidasius strain 12060. AB - The moderate thermophile, Bacillus thermoglucosidasius, transforms progesterone into four metabolites. These are 6alpha- and 6beta-hydroxyprogesterone, androstenedione and testosterone. This is the first report of bacterial 6alpha hydroxylation of steroids. The identity of the progesterone metabolites shows that there are three major types of transforming activity in this organism; C-17 C-20 lyase that cleaves the pregnane side chain of the substrate, C-17 oxidoreductase that interconverts the metabolites androstenedione and testosterone, and 6-hydroxylation. 6-hydroxylation activity was purified virtually to homogeneity and was shown to be catalysed by a cytochrome P-450 monooxygenase enzyme. This is the first report of a thermostable cytochrome P 450. PMID- 9877218 TI - Differential induction of glutathione S-transferase subunits by spironolactone in rat liver, jejunum and colon. AB - The effect of spironolactone pretreatment on glutathione S-transferase activity and on the relative content of the principal subunits (Ya, Yc, Yb1, Yb2 and Yp or 1, 2, 3, 4 and 7 respectively) was studied in rat liver, jejunum and colon. Male Wistar rats were injected with spironolactone i.p. at daily doses of 50, 100 and 200 micromol/kg body wt for 3 consecutive days. Glutathione S-transferase activities were assayed using 1-chloro-2,4-dinitrobenzene as substrate. Changes in subunit composition were evaluated by Western blot analysis in rats treated with the highest dose of spironolactone. The results demonstrated a dose dependent increase in enzyme activity in liver, while in jejunum the three tested doses exhibited the same magnitude of induction. No significant difference in glutathione S-transferase activity was observed between control and treated rats for the colon. Immunoblot analysis revealed more Ya and Yp protein in liver (140 and 118% increase respectively) and jejunum (45 and 145% increase respectively) from treated rats. While Ya and Yp relative contents were similar in jejunum, the latter subunit slightly contributed to total GST in liver, even in SL-treated animals. The inducer produced no change in subunit composition in colon. In conclusion, spironolactone was able to increase glutathione S-transferase activity mainly by induction of Ya subunit in liver and Yp subunit in jejunal mucosa, without affecting colonic enzyme. PMID- 9877220 TI - NMDA receptor antagonists enhance 5-HT2 receptor-mediated behavior, head-twitch response, in mice. AB - The purpose of this study was to characterize behavioral interactions between glutamatergic and serotonergic 5-HT2 receptors. Both competitive (AP-5 [D-2-amino 5-phosphovalerate] and D-CPP [3-(2carboxypiperazine-4yl)-propylphosphonate]) and noncompetitive (MK-801 [dizocilpine], ketamine, dextrorphan and dextromethorphan) N-methyl-D-aspartate (NMDA) receptor antagonists markedly enhanced a selective serotonergic behavior, the head twitch response (HTR), in mice. In contrast, NMDA itself inhibited 5-hydroxytryptamine (5-HT)-induced HTR in mice. These results suggest that glutamatergic neurotransmission may modulate serotonergic function at the 5-HT2 receptor. PMID- 9877219 TI - Tryptase mediates hyperresponsiveness in isolated guinea pig bronchi. AB - Hyperresponsiveness of airway smooth muscle to allergens and environmental factors has long been associated with the pathophysiology of asthma. Tryptase, a serine protease of lung mast cells, has been implicated as one of the mediators involved in the induction of hyperresponsiveness. As a consequence, tryptase inhibitors have become the subject of study as potential novel therapeutic agents for asthma. Secretory leukocyte protease inhibitor (SLPI) is a naturally occurring protein of human airways which exhibits anti-tryptase activity. To assess the potential therapeutic utility of SLPI in asthma, its effects were evaluated using in vitro and ex vivo models of airway hyperresponsiveness and compared with the effects of the small molecule tryptase inhibitor APC-366. Our results demonstrate that SLPI inhibits tryptase-mediated hyperresponsiveness in vitro and attenuates the hyperresponsiveness observed in airway smooth muscle from antigen-sensitized animals subjected to antigen exposure. The small molecule tryptase inhibitor APC-366 has a similar inhibitory effect. Thus, tryptase appears to be a significant contributor to the development of hyperresponsiveness in these models. To the extent that tryptase contributes to the development and progression of asthma, SLPI may possess therapeutic potential in this disease setting. PMID- 9877221 TI - Inhibitory effect of adrenomedullin (ADM) on the aldosterone response of human adrenocortical cells to angiotensin-II: role of ADM(22-52)-sensitive receptors. AB - Human adrenomedullin (ADM) is a 52-amino acid hypotensive peptide, which possesses a disulfide bridge-formed six-membered ring in 16-21 position. The ring structure, and both the N- and C-terminal amino-acid sequences seem to play a key role in the vascular effects of ADM(1-52), and we have investigated whether the same is true for the inhibitory effect of this peptide on the aldosterone response of zona glomerulosa (ZG) cells to angiotensin-II (ANG-II). Autoradiography showed the presence of abundant [125I]ADM(1-52) binding sites in the ZG of human adrenals, which were displaced not only by cold ADM(1-52), but also by both ADM(13-52) and ADM(22-52); ADM fragments 1-12, 15-22 and 16-31 were ineffective. ADM(1-52) and ADM(13-52), but not other fragments, concentration dependently inhibited ANG-II-stimulated aldosterone secretion of dispersed human adrenocortical cells. The aldosterone antisecretagogue actions of ADM(1-52) and ADM(13-52) were counteracted by ADM(22-52) in a concentration-dependent manner, while other ADM fragments were ineffective. In light of these findings the following conclusions could be drawn: (i) human ZG cells are provided with ADM(22 52)-sensitive receptors; (ii) the six-membered ring structure and the C-terminal, but not N-terminal, amino-acid sequence are both essential for ADM(1-52) to exert its antimineralocorticoid action; and probably (iii) the C-terminal sequence is needed for ADM(1-52) to bind its ZG receptors, while the ring structure is required for the receptor activation. PMID- 9877222 TI - Transepithelial transport and accumulation of flavone in human intestinal Caco-2 cells. AB - Flavonoids are found in many food items of plant origin. Intake of flavonoids has been linked to the prevention of human diseases including cancer. However, little is known about the intestinal absorption of flavonoids in the cellular level. This study was designed to study the absorption of dietary flavonoids using cultured human intestinal epithelial cell monolayer as a model system and 14C flavone as a model compound. 14C-flavone at 10 microM was found to move across the cell monolayer rapidly both from the luminal to basolateral direction and from the basolateral to luminal direction. The rate of transport from the luminal to basolateral direction was 5 times of the rate for phenylalanine, an aromatic amino acid. Flavone also accumulated substantially in the cells. Replacing sodium in the transport buffer with potassium did not affect the transport but reducing the incubation temperature significantly decreased the initial rate of transport. The presence of protein in the transport buffer reduced the initial rate of transport to half. Other flavonoids and hydrophobic chemicals at 100 microM had no effects on the transport. Together with the evidence from microscopic observation (Cancer Letts. 110: 41-48, 1996), this study supports that rapid diffusional transport may be the main route for flavonoid absorption. The ability of intestinal cells to accumulate flavone is consistent with the role of flavonoids in colon cancer prevention. PMID- 9877223 TI - Transplacental transfer and biotransformation studies of nicotine in the human placental cotyledon perfused in vitro. AB - Our objective was to study the characteristics of transfer and biotransformation of nicotine in the human term placenta. Nicotine transfer was studied by dually perfusing an isolated cotyledon of the human placenta in vitro. Nicotine metabolism to cotinine was investigated in intact tissue during perfusion and in placental microsomal fractions. Following the addition of nicotine (40 ng/ml) to the maternal side of the placenta, distribution into placental tissue (0.43 +/- 0.13 ng/ml/min) was three times higher than transfer to the fetal side of the placenta (0.15 +/- 0.01 ng/ml/min). The steady-state maternal-to-fetal transfer of nicotine was approximately 90% that of antipyrine (a marker of flow-dependent transfer). There was no evidence of nicotine metabolism to cotinine by intact placental tissue or in microsomal fractions. The observation that nicotine readily crosses the human placenta with no evidence of metabolism suggests that nicotine has the potential to cause adverse affects on the developing fetus. PMID- 9877224 TI - Evidence of a strong interaction of 2,4-dichlorophenoxyacetic acid herbicide with human serum albumin. AB - The interaction of 2,4-dichlorophenoxyacetic acid herbicide (2,4-D) with human serum albumin (HSA) was studied using fluorescence and differential scanning calorimetry (DSC). Fluorescence displacement of 1-anilino-8-naphtalenesulfonate (ANS) bound to HSA was used to evaluate the binding affinity of 2,4-D to HSA. The binding is associated to a high affinity site of HSA located in the IIIA subdomain. The association constant (Kass) of the herbicide was about 5 microM( 1), several times higher than the affinity found for pharmaceutical compounds. This relatively strong interaction with HSA was evidenced by the increase in HSA protein thermostability induced as consequence of herbicide interaction. 2,4-D induces an increase in the midpoint of thermal denaturation temperature from 60.1 degrees C in herbicide free solution to 75.6 degrees C in full ligand saturating condition. The calorimetric enthalpy and the excess heat capacity also increased upon 2,4-D binding. To investigate the possibility of other/s system/s of 2,4-D transport in blood, besides of HSA, the interaction of the herbicide with lipid monolayers was explored. No interaction was detected with any of the lipids tested. The overall results provided evidence that high affinity 2,4-D-HSA complex exhibits enhanced thermal stability and that HSA is the unique transport system for 2,4-D in blood. PMID- 9877225 TI - Streptozotocin-induced diabetes enhances protective effects of enalapril on nitric oxide-deficient stroke in stroke-prone rats. AB - Recently, we have shown that chronic administration of N-Nitro-L-Arginine Methyl Ester (L-NAME, an inhibitor of nitric oxide synthase) precipitates stroke in stroke-prone spontaneously hypertensive rats (SHRSP). Enalapril maleate, an angiotensin converting enzyme inhibitor was shown to delay the onset of such stroke. In the present study, five groups of 4-week-old SHRSP were used. Three groups of SHRSP were made diabetic using streptozotocin (100 mg/kg i.p.). One week later, the SHRSP from groups I (non-diabetic) and III (diabetic) chronically received L-NAME (0.5 g/L) in saline as drinking water. Two SHRSP groups, II (non diabetic) and IV (diabetic) received L-NAME (0.5 g/L) and enalapril maleate (20 mg/L) in saline as drinking water. Control SHRSP (group C; diabetic) received only saline to drink. SHRSP groups I and III developed stroke in 10+/-2 and 11+/ 2 days, respectively. The average stroke-free period in groups II and IV was 19+/ 2 and 28+/-2 days, respectively. Protective effect of streptozotocin-induced diabetes disappeared when SHRSP drinking L-NAME and enalapril, concurrently received subcutaneous injections of insulin (2 units daily per 100 g rat). Present data suggest that experimental diabetes delays the onset of L-NAME induced stroke in SHRSP only in the absence of angiotensin converting enzyme activity. In addition, diabetes-induced enhancement of stroke-protective effect of enalapril appears to be independent of reduction in mean and systolic blood pressures. PMID- 9877226 TI - Interspecies scaling of renally secreted drugs. AB - The objective of this study is to predict pharmacokinetic parameters (clearance, volume of distribution at steady state, and elimination half-life) in humans from animal data for drugs which are renally secreted in humans. Pharmacokinetic parameters of ten drugs were scaled-up from animal data obtained from the literature. Using simple allometry (pharmacokinetic parameter of interest vs body weight), total, renal and nonrenal clearances, volume of distribution and half life were predicted in humans. The predicted parameters were compared with the observed parameters. The results of the study indicated that it is likely that the predicted total and renal clearances from animal data will be underestimated in humans for renally secreted drugs. The prediction of renal clearance was improved by normalizing the renal clearance by a 'correction factor' for animals who exhibited renal secretion. The predicted volume and half-life were comparable with the observed values in man. Overall, the results of this study indicate that caution should be employed in interpreting the total and renal clearance of renally secreted drugs predicted by the allometric approach. PMID- 9877227 TI - Excessive activation of tyrosine kinases leads to inhibition of proliferation in a thyroid carcinoma cell line. AB - Autocrine stimulation of growth is a hallmark of many tumor cell lines. In this work we investigated the synthesis and secretion of growth factors and the expression of their corresponding receptors in HTC-TSHr thyroid carcinoma cells. These cells synthesize epidermal growth factor (EGF) receptors and platelet derived growth factor beta (PDGF beta) receptors and in addition transforming growth factor alpha (TGF alpha), PDGF-A and PDGF-B chains, respectively. Addition of EGF or PDGF-BB to the culture medium resulted in growth inhibition of HTC-TSHr cells. In contrast, treatment of the cells with low concentrations of neutralizing anti-TGF alpha antibodies or tyrosine kinase inhibitors led to stimulation of cell proliferation. Low concentrations of neutralizing anti-PDGF-B antibodies did not affect growth of the cells. As expected, cell proliferation was inhibited when high concentrations of either neutralizing anti-TGF alpha antibodies or anti-PDGF-B antibodies were applied. PDGF-AA did not influence growth of HTC-TSHr cells. We conclude that growth of HTC-TSHr thyroid carcinoma cells is influenced by two autocrine loops between TGF alpha and EGF receptors and between PDGF-B and PDGF beta receptors. However, our data suggest that excessive activation of tyrosine kinase receptors in these cells results in a relative inhibition rather than stimulation of growth. PMID- 9877228 TI - Nitric oxide enhances thyroid peroxidase activity in primary human thyrocytes. AB - Recent studies have demonstrated the production of the multi-functional messenger molecule nitric oxide (NO) by the thyroid gland. To examine a possible role for NO in thyroid function, we studied the acute and chronic effect of NO donors on thyroid peroxidase (TPO) activity in monolayer cultures of primary human thyrocytes, using a colorimetric assay technique. The presence of either S nitrosoglutathione (GS-NO) or sodium nitroprusside (SNP) (10(-6)-10(-4) M) at the time of the assay caused a significant increase in TPO activity. Pre-incubation of thyrocytes with 10(-5) M GS-NO for 3 days had no effect on the level of TPO activity when the assay was performed in the absence of NO donors. However, GS-NO pre-incubation significantly enhanced the acute stimulatory effect of GS-NO and SNP on TPO activity. These results suggest a possible role for NO in the regulation of TPO activity and thus thyroid hormone synthesis. PMID- 9877229 TI - Hydroxypropyl-beta-cyclodextrin and its combination with hydroxypropyl methylcellulose increases aqueous solubility of delta9-tetrahydrocannabinol. AB - Delta9-tetrahydrocannabinol (THC) is the main psychoactive constituent of Cannabis sativa L. and its therapeutic effects are currently under intensive study. However, THC has a very low aqueous solubility (1-2 microg/mL), which restricts its use as a pharmaceutical. The present study demonstrates that THC forms a drug-cyclodextrin complex in an aqueous solution of hydroxypropyl-beta cyclodextrin (HP-beta-CD), resulting in a thousand-fold increase in THC solubility. This improvement in solubility can be further increased by adding 0.1% hydroxypropylmethylcellulose to the HP-beta-CD solution. The present results suggest that the use of cyclodextrins might be a simple and useful method to overcome the poor water solubility of THC. PMID- 9877230 TI - Nobel Lecture. The identification of the sodium pump. AB - The identification of the sodium potassium pump as a Na+, K+ -ATPase is described. PMID- 9877231 TI - Purine nucleotide- and sugar phosphate-induced inhibition of the carboxyl methylation and catalysis of protein phosphatase-2A in insulin-secreting cells: protection by divalent cations. AB - Recently, we demonstrated that the 36 kDa catalytic subunit of protein phosphatase 2A (PP2Ac) undergoes methylation at its C-terminal leucine in normal rat islets, human islets and isolated beta cells; this modification increases the catalytic activity of PP2A [Kowluru et al. Endocrinology. 137:2315-2323, 1996]. Previous studies have suggested that adenine and guanine nucleotides or glycolytic intermediates [which are critical mediators in beta cell function] also modulate phosphatase activity in the pancreatic beta cell. Therefore, we examined whether these phosphorylated molecules specifically regulate the carboxyl methylation and the catalytic activity of PP2A in beta cells. Micromolar concentrations of ATP, ADP, GTP or GDP each inhibited the carboxyl methylation of PP2Ac and, to a lesser degree, the catalytic activity of PP2A. Likewise, the carboxyl methylation of PP2Ac and its catalytic activity were inhibited by [mono- or di-] phosphates of glucose or fructose. Additionally, however, the carboxyl methylation of PP2Ac was significantly stimulated by divalent metal ions (Mn2+ > Mg2+ > Ca2+ > control). The nucleotide or sugar phosphate-mediated inhibition of carboxyl methylation of PP2Ac and the catalytic activity of PP2A were completely prevented by Mn2+ or Mg2+. These data indicate that divalent metal ions protect against the inhibition by purine nucleotides or sugar phosphates of the carboxyl methylation of PP2Ac perhaps permitting PP2A to function under physiologic conditions. Therefore, these data warrant caution in interpretation of extant data on the regulation of phosphatase function by purine nucleotides. PMID- 9877232 TI - Subcellular localization and characterization of nucleoside diphosphate kinase in rat retina: effect of diabetes. AB - Nucleoside diphosphate kinase (NDP kinase) catalyzes the transfer of terminal phosphate from nucleotide triphosphates (e.g. ATP) to nucleotide diphosphates (e.g. GDP) to yield nucleotide triphosphates (e.g. GTP). Since guanine nucleotides play critical role(s) in GTP-binding protein (G-protein)-mediated signal transduction mechanisms in retina, we quantitated NDP kinase activity in subcellular fraction-derived from normal rat retina. A greater than 85% of the total specific activity was present in the soluble fraction, which was stimulated (up to 7 fold) by 2 mM magnesium. NDP kinase exhibited saturation kinetics towards di- and tri-phosphate substrates, and was inhibited by known inhibitors of NDP kinase, uridine diphosphate (UDP) or cromoglycate (CRG). We have previously reported significant abnormalities in the activation of G-proteins in streptozotocin (STZ)-diabetic rat retina (Kowluru et al. Diabetologia 35:624 631, 1992). Since NDP kinase has been implicated in direct interaction with and/or activation of various G-proteins, we quantitated both basal and magnesium stimulated NDP kinase activity in soluble and particulate fractions of retina derived from STZ-diabetic rats to examine whether abnormalities in G-protein function in diabetes are attributable to alterations in retinal NDP kinase. There was no effect of diabetes either on the basal or the magnesium-activated retinal NDP kinase activity. This study represents the first characterization of NDP kinase activity in rat retina, and suggests that in diabetes, this enzyme may not be rate-limiting and/or causal for the observed alterations in retinal G-protein functions. PMID- 9877233 TI - Evidence for nitric oxide mediated effects on islet hormone secretory phospholipase C signal transduction mechanisms. AB - We have investigated the putative role of nitric oxide (NO) as a modular of islet hormone release, when stimulated by the muscarinic receptor agonist phospholipase C activator, carbachol, with special regard to whether the IP3-Ca2+ or the diacylglycerol-protein kinase C messenger systems might be involved. It was observed that the NO synthase (NOS) inhibitor N(G)-nitro-L-arginine methylester (L-NAME) markedly potentiated insulin release and modestly inhibited glucagon release induced by carbachol. Similarly, insulin release induced by the phorbol ester TPA (protein kinase C activator) was markedly potentiated. Glucagon release, however, was unaffected. Dynamic perifusion experiments with 45C2+ loaded islets revealed that the inhibitory action of L-NAME on carbachol stimulated NO-production was reflected in a rapid and sustained increase in insulin secretion above carbachol controls, whereas the 45Ca2+ -efflux pattern was similar in both groups with the exception of a slight elevation of 45C2+ in the L-NAME-carbachol group during the latter part of the perifusion. No difference in either insulin release or 45Ca2+ -efflux pattern between the carbachol group and L-NAME-carbachol group was seen in another series of experiments with identical design but performed in the absence of extracellular Ca2+. However, it should be noted that in the absence of extracellular Ca2+ both 45Ca2+ -efflux and, especially, insulin release were greatly reduced in comparison with experiments in normal Ca2+. Further, in the presence of diazoxide, a potent K+ ATP-channel opener, plus a depolarizing concentration of K+ the NOS-inhibitor L-NAME still markedly potentiated carbachol-induced insulin release and inhibited glucagon release. The enantiomer D-NAME, which is devoid of NOS-inhibitory properties, did not affect carbachol-induced hormone release. TPA induced hormone release in depolarized islets was not affected by either L-NAME or D-NAME. The pharmacological intracellular NO donor hydroxylamine dose dependently inhibited insulin release stimulated by TPA. Furthermore, a series of perifusion experiments revealed that hydroxylamine greatly inhibited carbachol induced insulin release without affecting the 45Ca2+ -efflux pattern. In summary, our results suggest that the inhibitory effect of NO on carbachol-induced insulin release is not to any significant extent exerted on the IP3-Ca2+ messenger system but rather through S-nitrosylation of critical thiol-residues in protein kinase C and/or other secretion-regulatory thiol groups. In contrast, the stimulating action of NO on carbachol-induced glucagon release was, at least partially, connected to the IP3-Ca2+ messenger system. The main effects of NO on both insulin and glucagon release induced by carbachol were apparently exerted independently of membrane depolarization events. PMID- 9877234 TI - Blue light inhibits mitosis in tissue culture cells. AB - Irradiation of the mitotic (prophase and prometaphase) tissue culture PK (pig kidney embryo) cells using mercury arc lamp and band-pass filters postponed or inhibited anaphase onset. The biological responses observed after irradiation were: (i) normal cell division, (ii) delay in metaphase and then normal anaphase and incomplete cytokinesis, (iii) exit into interphase without separation of chromosomes, (i.v.) complete mitotic blockage. Cell sensitivity to the light at wavelengths from 423 and 488 nm was nearly the same; to the near UV light (wavelength 360 nm) it was 5-10 times more; to the green light (wavelength >500 nm) it was at least 10 times less. To elucidate the possible mechanism of the action of blue light we measured cell adsorption and examined cell autofluorescence. Autofluorescence of cytoplasmic granules was exited at wavelengths of 450-490 nm, but not at >500 nm. In mitotic cells fluorescent granules accumulated around the spindle. We suppose blue light irradiation induces formation of the free radicals and/or peroxide, and thus perturb the checkpoint system responsible for anaphase onset. PMID- 9877235 TI - UFT and leucovorin: a review of its clinical development and therapeutic potential in the oral treatment of cancer. AB - UFT is an oral antineoplastic drug combining uracil and tegafur in a 4:1 molar ratio. Tegafur acts as a prodrug of 5-fluorouracil (5-FU), being slowly metabolized by cytochrome P450 to 5-FU. Uracil competitively inhibits the metabolism of 5-FU, resulting in increased plasma and tumor 5-FU concentrations. At equimolar doses, higher peak plasma 5-FU concentrations are achieved with UFT plus oral leucovorin with similar systemic 5-FU exposure compared with low-dose continuous 5-FU infusions. The elimination half-life of 5-FU following UFT administration is approximately 7 h compared with 0.2 h with i.v. 5-FU. In phase II studies of UFT plus oral leucovorin for the treatment of advanced colorectal cancer, response rates ranged from 25 to 42%. UFT plus oral leucovorin is well tolerated, with manageable diarrhea being the only dose-limiting toxicity; the regimen is not associated with significant myelosuppression, mucositis, hand-foot syndrome or alopecia. UFT, with or without leucovorin, has also been evaluated alone or in combination with other cytotoxic agents for the treatment of advanced lung, breast and gastric cancers. UFT has also been evaluated as adjuvant therapy for colorectal, breast, gastric, head and neck, and superficial bladder cancers. UFT plus leucovorin offers patients an entirely oral cancer treatment, and appears to provide potential advantages over bolus 5-FU regimens with regard to toxicity and convenience of administration. These benefits should be advantageous in the adjuvant setting, as well as in advanced disease settings in which palliation is an important consideration. Ongoing clinical trials will further define the role of this promising oral treatment regimen. PMID- 9877236 TI - Treatments for newly diagnosed advanced ovarian cancer: analysis of survival data and cost-effectiveness evaluation. AB - The main therapeutic options currently available for patients with newly diagnosed advanced ovarian cancer include: (i) cisplatin-based chemotherapy at conventional doses without paclitaxel, (ii) paclitaxel+cisplatin at conventional doses and (iii) high-dose chemotherapy with autologous hematopoietic rescue. After conducting a literature search to identify large-scale clinical trials based on these three therapeutic modalities, we carried out an analysis of the survival data and evaluated the cost-effectiveness ratio where appropriate. Cost data were obtained from published information. Effectiveness was estimated by determining the values of mean lifetime survival (MLS). Our analysis included a total of 15 clinical trials. The values of MLS were 3.05 years per patient for cisplatin-based chemotherapy at conventional doses without paclitaxel (1931 patients), 2.95 years per patient for chemotherapy with paclitaxel+cisplatin at conventional doses (184 patients) and 5.76 years per patient for high-dose chemotherapy with autologous hematopoietic rescue (53 patients). As compared with cisplatin-based chemotherapy without paclitaxel, high-dose treatments with hematopoietic rescue yielded a significantly better survival. Using cisplatin based chemotherapy as a reference term, the incremental cost-effectiveness ratio for high-dose treatments was $25641 per life year gained (discounted dollars per discounted life year gained). Sensitivity testing suggested that the ratio remained below $50000 under most circumstances. We conclude that in the treatment of patients with advanced ovarian cancer, high-dose chemotherapy with hematopoietic rescue seems to be more effective and more cost-effective than standard treatments with cisplatin-based regimens at conventional doses. PMID- 9877237 TI - Pharmacokinetic-pharmacodynamic relationships between pirarubicin exposure and hematotoxicity: clinical application using only one blood sample. AB - The objective of the present study was to evaluate the relationship between the pharmacokinetic parameters of pirarubicin and of its metabolite doxorubicin measured in plasma and whole blood, and the hematologic toxicity of this drug, in order to evaluate the predictability of changes in white blood cells (WBC) by single measurement of drug concentrations. This pharmacokinetic-pharmacodynamic relationship was studied in a total of 45 patients with different tumor types treated by combined chemotherapy containing pirarubicin, administered as short infusion (10+/-2 min) at doses ranging from 50 to 90 mg. In 45 courses performed in 24 patients, we established the relationship between the half-product of pirarubicin level in whole blood at the end of the infusion and the duration of this infusion, which represents an estimate of the area under the time x concentration curve (AUC(PIRA,wb,ei) = C(PIRA,wb,ei) x duration of infusion/2), the age of the patients and the relative fall in WBC counts. These results allowed us to establish a predictive formula in order to anticipate the number of WBC that the patient will obtain about 12 days after treatment, at the nadir of the counting. WBCnadir = 0.032404 x Age + 2.005 + WBCinitial x e(-0.009316 x AUC(PIRA,wb,ei) + 4.202265), WBC being expressed as x 10(3) cells/microl and AUC(PIRA,wb,ei) in ng/ml x h. In a second step, the validation of the prediction was carried out in 43 courses from 21 patients treated in the same conditions, for which WBC(predicted nadir) was compared by linear regression to WBCcounted. We obtained a highly significant correlation: r = 0.656; p<0.0001). Therefore, we show in this paper that the hematological toxicity, especially the WBC nadir count, can be predicted from single-sample blood HPLC analysis. This rapid and easy prediction of leukopenia can help the clinician in anticipating important hematological toxicities and in deciding to start early prophylactic treatment with hematopoietic growth factors. PMID- 9877238 TI - Gemcitabine plus cisplatin combination given with amifostine (GAP) to heavily pretreated patients with gynecologic and peritoneal cancers: tolerance and activity in ovarian cancer. AB - Nine patients with cancers of gynecologic or peritoneal origin were treated with a combination of gemcitabine, amifostine and cisplatin (GAP). The rationale of including amifostine was primarily related to the amount of prior cisplatin the patients had received and the need to protect against additional neurotoxicity. After encouraging activity and tolerance had been noted, entry of three patients with severely compromised bone marrow was also allowed. These three patients required dose reductions and did not tolerate treatment more often than every other week, but nevertheless, one of them experienced a partial response lasting 9 months. Another two of the nine patients had CA125 decreases fulfilling Rustin's definition of response and one had elimination of ascites. Future studies of this combination are warranted. PMID- 9877240 TI - Intratumoral cisplatin administration in electrochemotherapy: antitumor effectiveness, sequence dependence and platinum content. AB - Electrochemotherapy using intratumoral cisplatin administration was tested on EAT tumors in mice. Mice were treated with eight electric pulses (100 micros, 1 Hz, 1040 V) and/or cisplatin (1, 2, 4 and 8 mg/kg). Cisplatin treatment resulted in up to 20 days of tumor growth delay. Electrochemotherapy resulted in tumor cures; local tumor control reached a plateau at 4 mg/kg in 67% of tumor cures. The maximal effect of electrochemotherapy was achieved when cisplatin was injected 5 min before or simultaneously with electric pulses application. Approximately two times more platinum was bound to DNA in electrochemotherapy than in cisplatin treated tumors at all time points tested. Our study shows that electrochemotherapy with intratumoral cisplatin administration is a very effective local treatment of EAT tumors with high curability rate. PMID- 9877241 TI - Microphysiometry: new technology for evaluation of anticancer drug activity in human tumor cells in vitro. AB - Microphysiometry is a non-invasive, physiological method where measurement of metabolic activity can be made on living human tumor cells. Indirect measurement of the extracellular acidification is measured over a pH-sensitive silicon membrane. In this study microphysiometry was employed for the study of cytotoxic agents used in therapy of cancer. Standard cytotoxic drugs with different postulated mechanisms of action were investigated using cell lines as well as primary cultures of patient tumor cells. Each investigated cytotoxic drug induced a characteristic pattern of metabolic activity. From these patterns, key features, like stimulation and inhibition of acidification, the time point when the response curves of the drugs fall below the control curve, and the maximum inhibition of acidification at 20 h, could be quantified. Most of the investigated drugs showed some initial stimulation of acidification rate during the experiments. For drugs producing a reduced metabolic rate at 20 h a concentration-response relationship was observed. The drug effects measured at 20 h were irreversible and correlated reasonably well with parallel measurements of membrane integrity using a standard cytotoxicity test. The results demonstrate the feasibility of 'on-line' measurements of metabolic activity using this approach and also revealed an unexpected variety of drug response profiles. PMID- 9877239 TI - Evaluation of ocular safety: tirapazamine plus cisplatin in patients with metastatic melanomas. AB - Ninety-six patients with metastatic melanoma treated with two consecutive tirapazamine-cisplatin combination chemotherapy regimens were followed for signs of therapy-related ocular toxicity. Baseline and follow-up data were obtained such that each patient acted as his own control. A battery of vision-related tests was performed. These included: best corrected visual acuity, color vision, retinal fundus examination and electro-oculograms (EOG). A brief health-related quality of vision test was administered at each follow-up visit to detect and evaluate self-perceived changes in visual status. In the first study, 48 patients received i.v. tirapazamine over 2 h at 260 mg/m2 (group 1) while in the second study 48 patients (group 2) received i.v. tirapazamine at 390 mg/m2. Visual system assessment was conducted at three timepoints: first at baseline, then at 6 weeks post-baseline, i.e. after two courses of chemotherapy and visit two upon discontinuation of therapy. There was no difference in visual acuity between group 1 and group 2 at baseline, follow-up 1 or at follow-up 2. Grouped data indicate that visual acuity was not affected by either dosage of chemotherapy. Group 1 at baseline found 15% below the normal EOG cutoff point, increasing to 23% at follow-up 1 and increasing at follow-up visit 2 to 33%. Group 2 demonstrated the same EOG findings, but the results were more magnified: baseline, 24%; follow-up 1, 44%; and follow-up 2, 44%. After eliminating those with abnormal color vision baselines, 21% (nine of 42) group 1 patients demonstrated abnormal color vision total error scores at follow-up 1 and 16.7% (four of 24) at follow-up 2. Few individuals showed changes in the higher dosage group. With the exception of one person in each dosage group, all changes were along the blue-yellow (tritan) axis, which is associated with acquired color defects. Of 96 patients examined, proven fundus changes were found in only four subjects. These fundus findings included retinal hemorrhages, retinal nerve fiber layer infarcts (cotton wool spots) and small retinal pigment epithelium detachments. There was no systematic statistical significant difference among the various measures of visual system outcome between groups or test times. Data from all tests for individual patients in both groups reveals a sporadic distribution of changes in visual system tests. If toxicity were pronounced, one would expect consistency in the findings and all or most of the assessment tests would be abnormal for a particular patient. However, patients who were abnormal on one measure of acuity were not necessarily abnormal on the other measures. PMID- 9877242 TI - Therapeutic effect of interstitial photodynamic therapy using ATX-S10(Na) and a diode laser on radio-resistant SCCVII tumors of C3H/He mice. AB - We examined the effect of interstitial photodynamic therapy (PDT) with a new photosensitizer ATX-S10(Na). This photosensitizer showed the strongest therapeutic effect 2-4 h after administration and was rapidly excreted from individual organs except tumor and liver 24 h after i.v. injection. Microscopic histofluorescent imaging showed fluorescence of ATX-S10(Na) in the cytoplasm of the tumor cells, but not in nuclei and in the vascular wall. Irradiation of Liniac 30 Gly+20 Gly slightly reduced the tumor size, but all mice died of relapse within 60 days after irradiation. In the PDT group, all tumors became non palpable and healing was achieved in 50% of mice 120 days after PDT. PMID- 9877243 TI - Antimitotic activity of diaryl compounds with structural features resembling combretastatin A-4. AB - Series of diaryl ethers, amines and amides have been synthesized and tested for antitumor activity. These diaryl compounds possess some of the structural features of combretastatin A-4 (a potent antimitotic agent). They were designed to discover whether transferring these structural motifs from stilbenes to heterosubstituted diaryl compounds would enhance their biochemical activities. Molecular modeling studies suggested that these diaryl compounds could adopt conformations similar to combretastatin A-4. However, although some agents (5-7) were cytotoxic and others (10 and 12) could interact with tubulin, none were as potent as combretastatin A-4. PMID- 9877244 TI - Antimetastatic effects of electrochemotherapy and of histoincompatible interleukin-2-secreting cells in the murine Lewis lung tumor. AB - Murine Lewis lung (3LL) tumors are characterized by the appearance of lung metastases after a regular period following their s.c. transplantation. We tested the respective efficiencies of various antitumor treatments: (i) electrochemotherapy (ECT), i.e. the systemic injection of bleomycin associated with electric pulses, locally delivered, that permeabilizes the tumor cells; (ii) intratumoral injection of histoincompatible cells that have been engineered in vitro to secrete high amounts of interleukin-2; and (iii) the combination of these two treatments. The growth of the s.c. transplanted tumors was followed up and the number of lung metastases was counted 14 days after the treatment. ECT alone resulted in the reduction of both the size of the tumor and the number of lung metastases. This latter effect can be partially explained by the effects of ECT on the s.c. tumor mass from which 3LL cells escape to colonize the lungs. The injection of IL-2-secreting cells alone had no effect on the s.c. mass and only a limited effect on the number of lung metastases. However, the combined treatment ECT plus IL-2-secreting cells resulted in antimetastatic effects potentiation that could result from stimulation of a non-specific immune response through an increase of NK activity. PMID- 9877245 TI - Human ovarian cancer xenografts in nude mice: chemotherapy trials with paclitaxel, cisplatin, vinorelbine and titanocene dichloride. AB - The new cytostatics titanocene dichloride and vinorelbine were compared to cisplatin and paclitaxel using a human ovarian cancer xenografts model. Biopsy material from a native human ovarian carcinoma was expanded and transplanted into 96 nude mice. The animals were divided into six treatment groups: cisplatin 3 x 4 mg/kg, paclitaxel 5 x 26 mg/kg, vinorelbine 1 x 20 mg/kg, titanocene dichloride 3 x 30 mg/kg, titanocene dichloride 3 x 40 mg/kg and a control group treated with 0.9% saline. Each experiment was repeated with eight mice in each treatment group. Treatment groups were evaluated in terms of average daily increase in tumor volume and average daily body weight increase of nude mice based on slopes of least-square regressions performed on individual animals. The slope factors alpha and beta of the body weight (alpha) and tumor volume changes (beta) within each group during the course of an experiment were calculated. Both a statistically significant decrease (p<0.05) in the body weight of the experimental animals (cisplatin: alpha = -0.5163, vinorelbine: alpha = -0.6598, paclitaxel: alpha = -0.6746, titanocene dichloride 3 x 30 mg/kg: alpha = -0.6259, titanocene dichloride 3 x 40 mg/kg: alpha = -0.7758) and a significant reduction (p<0.05) of the increase in tumor volume (cisplatin: beta = 12.049, vinorelbine: beta = 0.504, paclitaxel: beta = -1.636, titanocene dichloride 3 x 30 mg/kg: beta = 6.212, titanocene dichloride 3 x 40 mg/kg: beta= -0.685) was shown in all treated groups compared to the control group (alpha = -0.1398; beta = 23.056). No significant weight changes were observed between the individually treated groups. A statistically significant reduction of the tumor growth occured under paclitaxel (beta = -1.636), vinorelbine (beta = 0.504) and titanocene dichloride medication 3 x 40 mg/kg (beta = -0.685), as compared to the group treated with cisplatin (beta = 12.049). We found titanocene dichloride to be as effective as paclitaxel and more effective than cisplatin. Vinorelbine seems to be a very effective antineoplastic agent exhibiting a significant higher cytostatic effect than cisplatin. Both titanocene dichloride and vinorelbine provide new therapeutic options in women with ovarian carcinoma not responding to standard chemotherapy. PMID- 9877246 TI - Antitumor activity of tricyclic pyrone analogs, a new synthetic class of microtubule de-stabilizing agents, in the murine EMT-6 mammary tumor cell line in vitro. AB - Novel tricyclic pyrone (TP) analogs synthesized in Hua's laboratory (code names H10, H14 and H16) were tested against a spectrum of known antimitotic drugs for their ability to disrupt microtubule (MT) dynamics, alter the mitotic index, and prevent murine EMT-6 mammary sarcoma cells from synthesizing DNA and proliferating in vitro. At 2-10 microM, H10 inhibits DNA synthesis, tubulin polymerization and tumor cell growth to a greater degree than H14, whereas H16 has no effect. A linear skeleton with a pyridyl ring at C-3 of the A-ring, a pyran B-ring and no alkylation at C-7 of the C-ring is required for the antitumor activity of these TPs. Since H10 mimics the effect of vincristine (VCR), but not that of paclitaxel, on tubulin polymerization, TPs may represent a novel synthetic class of MT de-stabilizing anticancer drugs. H10 is less potent than VCR against tubulin polymerization (IC50: 1.5 microM versus 0.15 microM) and tumor cell proliferation (IC50: 1.5 microM versus 5 nM) but inhibits DNA synthesis (IC50: 10 microM) more effectively than all other MT-disrupting agents tested, except tubulozole-C. Although TPs disrupt DNA synthesis and might affect several phases of the cell cycle, the ability of H10 to increase the percentage of mitotic cells indicates that these novel compounds may be cell cycle-specific anticancer drugs useful for arresting mammalian cells in M-phase. PMID- 9877247 TI - Docetaxel and cyclophosphamide induced remission in platinum and paclitaxel refractory ovarian cancer. AB - Platinum-based chemotherapy is the standard treatment for advanced ovarian cancer, with response rates of 40-60%. In patients who fail platinum treatment, paclitaxel has resulted in response rates of 10-48%. Docetaxel has partial non cross-resistance with and is twice as potent in vitro as paclitaxel in inhibiting microtubule disaggregation. The combination of docetaxel and cyclophosphamide is synergistic in pre-clinical studies and clinically active in breast cancer. We present the case of a patient with platinum and paclitaxel refractory ovarian cancer who achieved a remission with docetaxel and cyclophosphamide. PMID- 9877248 TI - Stigmatization of obesity in medieval times: Asia and Europe. AB - OBJECTIVE: To describe the stigmatization of obesity in medieval times in Asia and in Europe. DESIGN: Literature review. RESULTS: Obesity was stigmatized in medieval Japan in part, at least, because it was viewed as the karmic consequence of a moral failing in a Buddhist context. The stigma in Europe was based on the Christian deadly sin of gluttony. CONCLUSION: Stigmatization of obesity is not new. It was foreshadowed centuries ago by attitudes deriving from then extant theories of the origins of obesity. PMID- 9877249 TI - Adipose tissue as an endocrine and paracrine organ. AB - The discovery of leptin has imparted great impetus to adipose tissue research by demonstrating a more active role for the adipocyte in energy regulation. Besides leptin, however, the adipose tissue also secretes a large number other signals. Cytokine signals, TNFalpha and IL-6, and components of the alternative pathway of complement influence peripheral fuel storage, mobilization and combustion, as well as energy homeostasis. In addition to the acute regulation of fuel metabolism, adipose tissue also influences steroid conversion and sexual maturation. In this way, adipose tissue is an active endocrine organ, influencing many aspects of fuel metabolism through a network of local and systemic signals, which interact with the established neuroendocrine regulators of adipose tissue. Thus, insulin, catecholamines and anterior pituitary endocrine axes interact at multiple levels with both cytokines and leptin. It may be proposed that the existence of this network of adipose tissue signalling pathways, arranged in an hierarchical fashion, constitutes a metabolic repertoire which enables the organism to adapt to a range of different metabolic challenges, including starvation, reproduction, times of physical activity, stress and infection, as well as short periods of gross energy excess. However, the occurrence of more prolonged periods of energy surplus, leading to obesity, is an unusual state in evolutionary terms, and the adipose tissue signalling repertoire, although sophisticated, adapts poorly to these conditions. Rather, the responses of the adipose tissue endocrine network to obesity are maladaptive, and lay the foundations of metabolic disease. PMID- 9877250 TI - Leptin resistance in obese humans: does it exist and what does it mean? PMID- 9877251 TI - Body mass index and percent body fat: a meta analysis among different ethnic groups. AB - OBJECTIVE: To study the relationship between percent body fat and body mass index (BMI) in different ethnic groups and to evaluate the validity of the BMI cut-off points for obesity. DESIGN: Meta analysis of literature data. SUBJECTS: Populations of American Blacks, Caucasians, Chinese, Ethiopians, Indonesians, Polynesians and Thais. MEASUREMENTS: Mean values of BMI, percent body fat, gender and age were adapted from original papers. RESULTS: The relationship between percent body fat and BMI differs in the ethnic groups studied. For the same level of body fat, age and gender, American Blacks have a 1.3 kg/m2 and Polynesians a 4.5 kg/m2 lower BMI compared to Caucasians. By contrast, in Chinese, Ethiopians, Indonesians and Thais BMIs are 1.9, 4.6, 3.2 and 2.9 kg/m2 lower compared to Caucasians, respectively. Slight differences in the relationship between percent body fat and BMI of American Caucasians and European Caucasians were also found. The differences found in the body fat/BMI relationship in different ethnic groups could be due to differences in energy balance as well as to differences in body build. CONCLUSIONS: The results show that the relationship between percent body fat and BMI is different among different ethnic groups. This should have public health implications for the definitions of BMI cut-off points for obesity, which would need to be population-specific. PMID- 9877252 TI - Hypothalamic expression of neuropeptide-Y in the New Zealand obese mouse. AB - OBJECTIVE: Increased levels of hypothalamic neuropeptide-Y (NPY) are thought to contribute to the manifestation of the obese phenotype. The aim of this study was to characterize the interactions between leptin, insulin and NPY in the pathogenesis of polygenic obesity. DESIGN AND METHODS: A polygenic obese animal model, the New Zealand obese mouse (NZO) and its age-matched control, was used to assess the hypothalamic mRNA expression of NPY, the insulin receptor (IR) and the leptin receptor (Ob-Rb), by semiquantitative polymerase chain reaction. Experiments were performed early (at eight weeks old) and late (at 40 weeks old) in the life of these animals. RESULTS: Serum glucose was significantly elevated in obese mice. Serum insulin levels were not different between obese and lean mice, whereas serum leptin levels were significantly elevated in obese mice and increased continuously during life [reaching extremely high values at 40 weeks (41.5+/-4.1 vs 3.4+/-0.25 ng/ml for obese and lean, respectively). The hypothalamic expression of NPY mRNA was significantly higher in NZO mice compared to controls at both eight weeks (2.3-fold) and 40 weeks (1.9-fold), respectively, whereas expression of IR and Ob-Rb remained unaffected. CONCLUSION: Increased hypothalamic expression of NPY due to leptin resistance, may be involved in the development of polygenic obesity. Unchanged Ob-Rb expression suggests that either a defective hypothalamic uptake or defects in Ob-Rb signalling underly this process. PMID- 9877253 TI - Influence of alcohol consumption and various beverages on waist girth and waist to-hip ratio in a sample of French men and women. AB - OBJECTIVE: The goal of the present study was to assess the association between alcohol consumption and abdominal fat deposition in France, a country where wine is the most commonly consumed alcoholic beverage. METHODS: We analyzed the association between alcohol consumption and various markers of body weight, in a population sample recruited in three distinct geographical areas of France (MONICA centers). This sample included men (n = 1778) and women (n = 1730) aged 35- 64 y, randomly selected from electoral rolls. Alcohol consumption was assessed with a quantitative frequency questionnaire. RESULTS: Alcohol intake ranged from 0-1655 ml of alcohol per week. Wine was the main source of alcohol, representing 67% of total alcohol intake in both genders. In men, there was no association between alcohol intake and body mass index (BMI) or body weight, whereas in women, alcohol consumption was inversely correlated with BMI (P < 0.0001) and body weight (P < 0.0002). In men, total alcohol consumption was positively associated with waist-to-hip ratio (WHR, P < 0.0002) and waist girth (P < 0.004), independently of BMI. Similarly, in women, positive associations were found between alcohol intake and WHR (P < 0.0001) as well as waist girth (P < 0.0001), independently of BMI. In a linear regression model including types of beverages and usual confounders, reporting consumption of either wine, beer or spirit was poorly associated with WHR in men, whereas wine (P < 0.0008) and beer (P < 0.0001) consumptions were both positively associated with WHR in women. However, there was no evidence of a statistically significant heterogeneous effect of wine, beer and spirits on WHR in both genders. CONCLUSION: In a sample of representative French people, in whom wine is the most common alcoholic beverage, alcohol consumption is associated with greater WHR independently of BMI in both men and women. PMID- 9877254 TI - The interactions between hypothalamic-pituitary-adrenal axis activity, testosterone, insulin-like growth factor I and abdominal obesity with metabolism and blood pressure in men. AB - OBJECTIVE: To examine potential interactions between abdominal obesity, endocrine, metabolic and hemodynamic perturbations. SUBJECTS: A subgroup of 284 men from a population sample of 1040 at the age of 51 y. MEASUREMENTS: Anthropometric measurements included body mass index (BMI, kg/m2), waist/hip circumference ratio (WHR) and abdominal sagittal diameter (D). Endocrine measurements were a modified, low dose (0.5 mg) dexamethasone suppression test (Dex), testosterone (T) and insulin-like growth factor I (IGF-I). Overnight fasting values of blood glucose, serum insulin, triglycerides, total, low and high density lipoprotein cholesterol, as well as resting heart rate and blood pressure were also determined. RESULTS: Arbitrary subdivisions of the men were performed to obtain subgroups of low T and IGF-I values (lowest decile, borderlines < or =13.13 nmol/I and < or =128.80 microg/l, respectively) and normal or blunted Dex. Significant relationships with BMI, WHR or D, and abnormal metabolic and hemodynamic factors, usually with the exception of total and low density lipoprotein cholesterol, were then found in subgroups with different endocrine profiles. These included men with a blunted Dex test with low T or IGF I values, as well as men with a normal Dex test and low or normal T or IGF-I values. In addition, a group with isolated low Dex suppression, as well as another group without endocrine abnormalities, showed such relationships. These findings suggest that, in men, obesity factors are associated with metabolic and hemodynamic complications with or without the presence of perturbations of hypothalamic-pituitary-adrenal axis (HPA) regulation or low T or growth hormone secretion. In order to generate hypotheses concerning the nature of the impact of the endocrine perturbations in abdominal obesity and its metabolic complications, path analyses were performed, testing different models. These models included the endocrine measurements (Dex test, T and IGF-I), the WHR and D (representing abdominal distribution of fat), BMI (representing obesity), as well as insulin and triglyceride values (representing metabolic perturbations). The results showed a satisfactory fit (goodness-of-fit index: 0.945 - 1.0) for the path diagrams: Dex --> T/IGF-I --> WHR or D --> insulin --> triglycerides with additional direct input of blunted Dex on insulin values (see Figure 1). With BMI as determinant, essentially the same results were found with the addition of a direct pathway between Dex and BMI as well as between IGF-I-T and insulin (Figure 2). There was no evidence for pathways where WHR or BMI determined endocrine variables. CONCLUSIONS: The results suggest that abdominal obesity with or without endocrine abnormalities exerts a major impact on abnormalities in metabolic and hemodynamic variables. Abdominal obesity seems to be dependent on endocrine abnormalities, which in turn show direct or indirect relationships to the metabolic and circulatory variables, including a direct pathway between HPA axis perturbations and accumulation of total body fat as indicated by the BMI. It is therefore suggested that endocrine perturbations are followed by obesity and by storage of an elevated proportion of fat in visceral depots, followed by metabolic and hemodynamic abnormalities. This is statistical evidence which is supported by evidence of mechanistic links in previous studies, suggesting the possibility of causal relationships. The results also indicate subgroups of abdominal obesity and its associated metabolic and hemodynamic abnormalities, which might be due to the input of different pathogenetic factors. PMID- 9877255 TI - Fasting serum leptin levels in the analysis of body mass index cut-off values: are they useful for overweight screening in children and adolescents? A school population-based survey in three provinces of central Italy. AB - OBJECTIVE: Body mass index (BMI) was determined in a population of school students from three provinces of central Italy. Fasting serum leptin concentrations were assayed in a large number of subjects from the same area, to determine their distribution as plotted against the standard deviation score (z score) of BMI. DESIGN, SUBJECTS AND MEASUREMENTS: Height and weight were recorded from 31170 subjects (16175 male and 14995 female), aged 3-18 y, to construct BMI charts of children and adolescents from central Italy. Percentiles and z-score were calculated using the LMS method of Cole. Serum leptin concentrations were assayed in 1929 subjects (996 male and 933 female) after overnight fasting. RESULTS: BMI percentiles of central Italy were higher than those from standards of other European and USA populations. When plotted against the z-score of BMI, serum leptin values were distributed according to an exponential curve, showing a steep pattern and a wide distribution, as BMI values increased. The hypothesis of the existence of two subgroups, based on a different relation between leptin and BMI, was verified and a separation point between the two subgroups was identified using cluster analysis, discriminant analysis and a novel method developed by our group, hereafter referred to as 'regression clustering'. This method allows identification of the value of the independent variable (z-score of BMI) which can be taken as a separation point. This analysis provided the best results and indicated the following separation points: central Italy standard, z-score = 0.72 (76.4th percentile) for males and z-score = 0.69 (75.5th percentile) for females; French standard (the one suggested for a European population by the European Childhood Obesity Group, ECOG), z-score = 1.46 (92.8th percentile) for males and z-score = 1.96 (97.5th percentile) for females. Similar but variable results were obtained when the same analysis was performed on serum leptin concentration, subdivided according to pubertal development (stage I, stage II-III, stage IV-V). CONCLUSIONS: Children and adolescents from central Italy had greater BMI percentiles when compared to other European populations. Fasting serum leptin concentrations showed a distribution pattern related to z-score, thus allowing to identification of two different subgroups. The z-scores of BMI, identified as separation points, indicated a trend to leptin production by adipocytes that could be taken as indicators of significant increases of fat mass. This study proposes criteria and a statistical approach that could be useful in the identification of BMI cut-off values when screening children and adolescents for overweight. PMID- 9877256 TI - Relationships between an index of body fat distribution (based on waist and hip circumferences) and stature, and biochemical complications in obese children. AB - OBJECTIVE: To determine the clinical utility of a new age-adjusted measure of body fat distribution (based on waist and hip circumferences) and stature, in relation to biochemical complications in obese children. DESIGN: Cross-sectional, clinical study. The formula to calculate the common standard deviation score (SDS) of waist-to-hip ratio/height (WHR/Ht) was obtained from the data of control children. The relationship between WHR/Ht SDS, as the age-adjusted measure, in obese children and their clinical laboratory data was evaluated. SUBJECTS: Outpatient obese Japanese children (102 boys and 75 girls) and control children (508 boys and 549 girls), ranging in age from 6-15 y. MEASUREMENTS: Height, body weight, waist girth, hip girth, triceps and subscapular skinfold thicknesses, as anthropometric measures. Percent overweight, percent body fat, waist girth, WHR and WHR/Ht SDS as criteria for obesity. Clinical laboratory analyses for fasting blood samples of obese children. RESULTS: The WHR/Ht SDS closely correlated with age in obese children, thus reflecting the progress of abdominal obesity during growth. The obese boys were more hyperlipidaemic than the girls were, although the percent overweight was similar in both genders. The percent overweight, percent body fat, waist girth and WHR/Ht SDS all correlated well with triglyceride (TG), alanine aminotransferase (ALT) and insulin in boys, whereas only waist girth and WHR/Ht SDS showed a close correlation with TG and insulin in girls. The obese subjects were subdivided according to the number of abnormal values observed in TG, ALT and insulin. For obese boys, all five indices of obesity were higher in the groups with complications than in the group without. In the girls, only the WHR/Ht SDS constantly differed between subgroups. WHR/Ht SDS most obviously distinguished the groups with complications from the other group with a wide margin of difference (2-fold in boys and > 2-fold in girls) in the mean values. CONCLUSION: The WHR/Ht SDS can serve as an index predicting the occurrence of biochemical complications in obese children ranging from the age of 6-15 y. PMID- 9877257 TI - Role of behaviour modification in the treatment of childhood obesity with the parents as the exclusive agents of change. AB - OBJECTIVE: This study examined the reduction in overweight and changes in eating related behaviours in obese children treated with a family-based approach, in which the parents were the exclusive agents of change. Results were compared to the conventional approach in which children are responsible for their own weight loss. DESIGN: A one-year longitudinal prospective design was used. Participants were randomly allocated for the experimental (parents as the agents of change) or the conventional (children as the agents of change) intervention. METHODS: Sixty obese children (20% over ideal weight for age, height and gender) aged 6-11 y (mean+/-s.d. 9.2+/-1.0 y) were included in the study. Anthropometric measurements and biochemical tests were performed on the children and their parents before and after the programme, and both parents completed a sociodemographic and a family eating and activity habits questionnaire. Hour-long support/educational sessions were conducted by a clinical dietitian-14 sessions for the parents in the experimental intervention and 30 for the children in the conventional intervention. Individual sessions were held for members of both groups, when necessary. RESULTS: Significant differences were found between the two groups in the reduction of exposure to food stimuli and changes in eating habits (eating while standing, watching TV, reading or doing homework, eating following stress and eating between meals). Mean weight reduction (by percentile) was significantly greater in the experimental intervention group (parent-only treatment) in comparison to the conventional intervention group (child-only treatment). CONCLUSIONS: Treatment of childhood obesity with the parents as the exclusive agents of change, induces more behavioural changes as well as greater weight loss, than the conventional approach. PMID- 9877259 TI - What is "obesity"--an analysis of referral letters to an obesity unit. AB - OBJECTIVES: To evaluate the quality of referral letters concerning obese patients from general practice and other specialities to an obesity unit. DESIGN: Retrospective analysis of referral letters in 500 consecutive patient records. SETTING: Academic specialist obesity unit. RESULTS: Most letters came from general practice (70%), followed by internal medicine/endocrinology (12%). Information on body weight was missing in 13% of all referrals and on height in 24%. Waist circumference was mentioned in 1%. Relevant data substantiating possible comorbidities, in particular the metabolic syndrome, was missing in 92 97%. Relevant medication was indicated in 22% of the referrals. On a 3-point, graded global evaluation scale of the referral quality, only 7% of all letters were found to be 'ideal'. CONCLUSION: The low professional quality of these referrals may reflect the fact that physicians find the term 'obesity' enough to warrant the referral without further specifications. An alternative explanation for the inadequate referrals is the well documented negative attitude of physicians, who consider obesity as sloth and as a self-inflicted condition, not necessitating further medical details. PMID- 9877258 TI - Effect of adrenalectomy on the slimming activity of liposome-carried oleoyl estrone in the rat. AB - OBJECTIVE: To determine the extent of glucocorticoid counter-regulatory control in the slimming action of oleoylestrone. DESIGN: Control and adrenalectomized rats were subjected to a seven-day treatment with 3.5 micromol/kg/d oleoylestrone in liposomes injected i.v. continuously by implanted osmotic minipumps. SUBJECTS: Sham-operated control and adrenalectomized lean Zucker rats. MEASUREMENTS: Body weight and food intake; plasma glucose, urea, insulin, leptin and corticosterone; liver glycogen. RESULTS: Treatment with oleoyl-estrone resulted in decreases in body weight and in food intake, as well as in circulating glucose, insulin and leptin. Combined adrenalectomy and oleoyl-estrone treatment resulted in a loss of almost 15% body weight in only seven days, with a severe drop in circulating glucose and insulin, almost total disappearance of plasma leptin and liver glycogen and a 3-fold rise in circulating urea. Food intake decreased sharply, which resulted in the exhaustion of energy reserves. CONCLUSION: The results presented here, strongly support the hypothesis that glucocorticoids play an important role in the modulation of oleoyl-estrone-induced imbalance of energy intake and expenditure. The large effect of oleoyl-estrone on glucose, glycogen- and protein-derived (urea levels) energy in adrenalectomized rats, provides more evidence for the assumed protective role of glucocorticoids against the oleoyl estrone-induced net loss of energy reserves. The results also show the powerful destabilizing effects of unchecked oleoyl-estrone on energy balance. PMID- 9877260 TI - An increase of circulating leptin in patients with liver cirrhosis. AB - OBJECTIVE: Leptin, the ob gene product, is an anorexigenic peptide secreted from adipose tissue. However, the mechanism of leptin clearance/degradation has not been well determined in humans. The present study was undertaken to examine a possible involvement of liver in determining circulating leptin concentrations in humans. SUBJECTS: In the present study 58 healthy control subjects and 68 patients with liver cirrhosis (LC) without any renal dysfunction were randomly included. METHOD: The serum immunoreactive leptin (IRL) concentrations relative to the body mass index (BMI) were determined. Serum IRL and estradiol (E2) concentrations were assayed by radioimmunoassay (RIA). RESULTS: The correlations between the BMI and circulating IRL concentrations were all significant in male healthy controls (M-C), male patients with LC (M-LC), female healthy controls (F C) and female patients with LC (F-LC). Circulating IRL concentrations were significantly higher than control in F-LC but not M-LC groups. The ratio of circulating IRL concentrations to the BMI was significantly higher in the M-LC group than in the M-C group and also significantly higher in the F-LC group than in the F-C group. The correlation between the IRL/BMI ratio and serum total bilirubin concentrations was significant (r = 0.417, P < 0.05) in the M-LC group, but not in the F-LC group. There was no significant correlation of the IRL/BMI ratio to serum E2 or albumin concentrations in either M-LC or F-LC groups. CONCLUSION: The present data demonstrated that the rate of increase in circulating IRL concentrations with the BMI was higher in LC patients of both genders. Liver may play a role in determining circulating leptin levels. PMID- 9877261 TI - Plasma concentrations of soluble TNF-alpha receptors in obese subjects. AB - BACKGROUND: Recent studies show an increased adipose production of tumor necrosis factor-alpha (TNF-alpha) in human obesity. It was hypothesized from this finding and other data, that TNF-alpha may be a mediator of obesity-linked insulin resistance. OBJECTIVE: The aim of this study was to measure plasma concentrations of the two soluble TNF-alpha receptors, together with those of TNF-alpha in subjects with severe obesity with and without type 2 diabetes mellitus, in comparison to a lean control group, to examine whether plasma concentrations reflect an up-regulation of the TNF system in adipose tissue. PATIENTS AND METHODS: Plasma concentrations of the two soluble TNF-alpha receptors were measured in 49 obese subjects (mean body mass index (BMI): 44.9 kg/m2, 95% confidence intervals (CI) 42.3-47.5 kg/m2, including 19 type 2 diabetic individuals) and 28 lean controls, by using a highly sensitive enzyme-linked immunoassay (ELISA) technique. TNF-alpha concentrations were determined in 28 obese (10 with diabetes) and 23 lean subjects. RESULTS: The obese subjects showed significantly higher plasma concentrations of the soluble p60 and p80 TNF receptor, respectively, compared to the lean control group, independent of the presence of diabetes. Multiple regression analysis, with the p80 TNF receptor as dependent variable, revealed that BMI and log insulin significantly affected the plasma concentration of this soluble receptor subtype, explaining 46% of the variance, whereas for the p60 TNF receptor, only BMI turned out to influence plasma concentrations. TNF-alpha plasma concentrations were not different between the three groups (Kruskal-Wallis test: P=0.34), but due to the low power of the test, an effect of obesity on TNF-alpha is not excluded. CONCLUSION: These data indicate that plasma concentrations of both soluble TNF receptors are elevated in obesity and insulin resistance, possibly as a function of excess body fat. The reported adipose overexpression of TNF-alpha does not seem to be reflected by elevated plasma concentrations, suggesting a primarily local role of the cytokine. PMID- 9877262 TI - Influence of the A-->G (-3826) uncoupling protein-1 gene (UCP1) variant on the dynamics of body weight before and after gastroplasty in morbidly obese subjects. PMID- 9877263 TI - Intensified conditioning regimen in bone marrow transplantation for Philadelphia chromosome-positive acute lymphoblastic leukemia. AB - We investigated an intensified conditioning regimen including fractionated total body irradiation (12 Gy), etoposide (30-45 mg/kg) and cyclophosphamide (120 mg/kg), followed by autologous (n = 5), allo-related (n = 13) or allo-unrelated (n = 6) bone marrow (n = 22) or peripheral stem cell (n = 2) transplantation in patients with Philadelphia chromosome-positive acute lymphoblastic leukemia. One patient received busulfan (16 mg/kg) instead of TBI. Nineteen patients were transplanted in 1CR, two in 2CR, one in 1PR and two in relapse. Major toxicity was mucositis grade II according to the Bearman scale in all patients. The treatment-related mortality was 25%, mainly due to infection or GVHD after allogeneic transplantation. After a median follow-up of 45 months (range 2-93), nine patients (37.5%) remain alive in CR. Nine patients (37.5%) relapsed and eight (33.3%) of these subsequently died. After autologous transplantation, four of five patients (80%) relapsed and died. Late relapse was seen after allogeneic, as well as autologous transplantation, at 33 and 59 months, respectively. The Kaplan-Meier estimate of leukemia-free survival for all patients is 38% at 3 years (95% CI: 18-58%) and 35% at 5 years (95% CI: 15-55%). For allogeneic transplants in first CR (n = 15) the estimate of disease-free survival was 46% at 3 years (95% CI: 19-73%) and 34% at 5 years (95% CI: 17-51%). Patients aged below 30 years had a better estimated overall survival at 3 years (61% vs 11%, P < 0.001). The bcr-abl fusion transcript (p210 vs p190 vs p210/190) did not affect disease-free or overall survival. In our experience, an intensified conditioning regimen seems to improve the results of bone marrow transplantation in patients with Ph+ acute lymphoblastic leukemia. However, the high relapse rate warrants novel approaches to enhance anti-leukemic efficacy. PMID- 9877264 TI - A randomised, blinded, placebo-controlled, dose escalation study of the tolerability and efficacy of filgrastim for haemopoietic stem cell mobilisation in patients with severe active rheumatoid arthritis. AB - Autologous haemopoietic stem cell transplantation (HSCT) represents a potential therapy for severe rheumatoid arthritis (RA). As a prelude to clinical trails, the safety and efficacy of haemopoietic stem cell (HSC) mobilisation required investigation as colony-stimulating factors (CSFs) have been reported to flare RA. A double-blind, randomised placebo-controlled dose escalation study was performed. Two cohorts of eight patients fulfilling strict eligibility criteria for severe active RA (age median 40 years, range 24-60 years; median disease duration 10.5 years, range 2-18 years) received filgrastim (r-Hu-methionyl granulocyte(G)-CSF) at 5 and 10 microg/kg/day, randomised in a 5:3 ratio with placebo. Patients were unblinded on the fifth day of treatment and those randomised to filgrastim underwent cell harvesting (leukapheresis) daily until 2 x 10(6)/kg CD34+ cells (haemopoietic stem and progenitor cells) were obtained. Patients were assessed by clinical and laboratory parameters before, during and after filgrastim administration. RA flare was defined as an increase of 30% or more in two of the following parameters: tender joint count, swollen joint count or pain score. Efficacy was assessed by quantitation of CD34+ cells and CFU-GM. One patient in the 5 microg/kg/day group and two patients in the 10 microg/kg/day group fulfilled criteria for RA flare, although this did not preclude successful stem cell collection. Median changes in swollen and tender joint counts were not supportive of filgrastim consistently causing exacerbation of disease, but administration of filgrastim at 10 microg/kg/day was associated with rises in median C-reactive protein and median rheumatoid factor compared with placebo. Other adverse events were well recognised for filgrastim and included bone pain (80%) and increases in alkaline phosphatase (four-fold) and lactate dehydrogenase (two-fold). With respect to efficacy, filgrastim at 10 microg/kg/day was more efficient with all patients (n = 5) achieving target CD34+ cell counts with a single leukapheresis (median = 2.8, range = 2.3-4.8 x 10(6)/kg, median CFU-GM = 22.1, range = 4.2-102.9 x 10(4)/kg), whereas 1-3 leukaphereses were necessary to achieve the target yield using 5 microg/kg/day. We conclude that filgrastim may be administered to patients with severe active RA for effective stem cell mobilisation. Flare of RA occurs in a minority of patients and is more likely with 10 than 5 microg/kg/day. However, on balance, 10 microg/kg/day remains the dose of choice in view of more efficient CD34+ cell mobilisation. PMID- 9877265 TI - Autologous bone marrow transplantation with monoclonal antibody purged marrow for children with acute lymphoblastic leukemia in second remission. Spanish Working Party for BMT in Children. AB - The purpose of this study was to evaluate the outcome of children with acute lymphoid leukemia (ALL) in second remission who have undergone high-dose chemotherapy and radiotherapy and autologous bone marrow transplantation (ABMT) with monoclonal antibody purged marrow, and to determine the main prognostic factors. From 1987 to 1992, 55 children with ALL in second remission underwent ABMT. The conditioning regimen consisted of total body irradiation (TBI) plus cyclophosphamide in 21 patients and TBI plus cyclophosphamide plus cytarabine or VP-16 in 28 patients; the remaining six patients were treated with chemotherapy alone (cyclophosphamide and busulfan, and/or VP-16). The marrow was purged using monoclonal antibodies and complement or magnetic microspheres in all cases. All patients engrafted. Three patients (5%) died early post transplant from infections. Twenty-six patients (47%) relapsed (median 150 days); 26 patients (47%) are alive and in complete remission (CR) at a median of 36 months. The Kaplan-Meier estimation showed a probability of event-free survival (EFS) of 46 +/- 0.007%. In the univariate analysis, first CR length and conditioning with TBI plus two or more cytotoxic drugs were found to be the most significant predictors of EFS. ABMT with purged marrow is a treatment modality which offers a chance of cure in children with ALL after relapse, including children who relapse early. PMID- 9877266 TI - Helper and cytotoxic T cell precursor frequencies are not predictive for development of acute graft-versus-host disease after partially T cell-depleted HLA-identical sibling BMT. AB - Despite the use of partially T cell-depleted grafts, 20% of the recipients of an HLA-identical sibling marrow graft develop aGVHD > or = II. This indicates that the current method for selecting a sibling donor, ie serological typing for HLA A, B and DR, and a mixed lymphocyte culture (MLC) or molecular typing for HLA DRB/DQB, is not predictive for aGVHD. In order to optimise our selection procedure, we retrospectively analysed patients who developed aGVHD > or = II by means of sequencing based typing for HLA-DPB and frequency analysis of alloreactive helper and cytotoxic T lymphocyte precursors (HTLp-f and CTLp-f). Patients who did not develop aGVHD or developed aGVHD grade I served as controls. Retrospective typing for HLA-DPB revealed only a single disparity in the group with aGVHD > or = II, indicating that mismatches for antigens other than HLA are the major cause of aGVHD in these patients. Furthermore, in our patient group, neither HTLp-f nor CTLp-f were predictive for development of aGVHD indicating that these assays in their current set-up are insufficiently sensitive to predict aGVHD in BMT with a partially T cell-depleted graft. We conclude, that HLA identical siblings can be identified by means of serological typing for HLA-A and B and intermediate resolution molecular typing for DRB and DQB, but that for the prediction of aGVHD cellular tests with higher sensitivity and specificity as compared to the currently used HTLp-f and CTLp-f assays need to be developed. PMID- 9877267 TI - Donor leukocyte infusions for recurrent hematologic malignancies after allogeneic bone marrow transplantation: impact of infused and residual donor T cells. AB - We evaluated the efficacy and toxicity of different doses of donor T cells given with donor leukocyte infusions (DLI) as treatment for relapse of various hematologic malignancies after allogeneic bone marrow transplantation (BMT). We also studied whether DLI treatment was more effective if circulating T cells were exclusively of donor origin (complete donor T cell chimeras) as compared with T cells originating from both donor and recipient (mixed T cell chimeras). Twenty eight patients were studied of whom 24 had a complete donor T cell chimerism. The malignancies were as follows: chronic myeloid leukemia (CML) in chronic phase (CP) (n = 9); more advanced CML (n = 5); multiple myeloma (MM) (n = 5); acute leukemia (AL) (n = 9). T cell doses varied from 0.1 x 10(7) to 33 x 10(7) T cells/kg. Eight patients received two to four DLI courses because they failed to respond to one course. Thirteen of 14 patients with CML, including four patients with more advanced CML, achieved complete remission (CR). All five patients with MM responded, including three CRs. Six patients (three with CML, three with MM) responded only after two to four DLI courses. Patients with CML-CP were likely to respond to as few as 1 x 10(7) T cells/kg whereas patients with MM generally responded when they received > or = 10 x 10(7) T cells/kg. However, despite the infusion of high T cell doses (up to 32 x 10(7) T cells/kg), practically all patients with AL failed to respond. The likelihood of response was strongly related to the occurrence of graft-versus-host disease (GVHD) in patients with CML and MM (P = 0.0002), although GVHD was not helpful for patients with AL. Higher T cell doses (> or = 10 x 10(7)/kg) induced serious GVHD (n = 17) and marrow aplasia (n = 5), and GVHD was directly or indirectly the cause of death for six patients. Finally, there were no obvious differences in responses between complete donor T cell chimeras and mixed T cell chimeras. PMID- 9877268 TI - Autologous peripheral blood stem cell transplantation in acute myeloblastic leukaemia and myelodysplastic syndrome patients: evaluation of tumour cell contamination of leukaphereses by cytogenetic and molecular methods. AB - We evaluated 18 acute myeloblastic leukaemia (AML) and myelodysplastic syndrome (MDS) patients with abnormal karyotype at diagnosis who underwent peripheral blood stem cell (PBSC) transplantation. To evaluate the presence of residual tumour cells, bone marrow (BM) samples and PBSC collections were analysed by cytogenetics and in selected cases also by fluorescence in situ hybridisation (FISH) and molecular studies. All patients were considered to be in morphologic and cytogenetic complete remission (CR) at the time of mobilisation. Seven patients showed neoplastic cells in PBSC harvest and/or BM specimen before reinfusion. Cytogenetic studies revealed contamination in apheretic collections in one patient only, while three patients had BM but not PBSC contamination. Three more patients had leukaemic cells both in the BM and PBSC. All but one (with only BM contamination) of these patients relapsed within 9 months. However, five more patients relapsed after transplantation: in four cases there was no cytogenetic sign of contamination either in PBSC or BM cells and in one case no molecular evidence was revealed either. This study suggests that, whereas the presence of leukaemic cells in autologous grafts correlates with a poor prognosis, the lack of detection of tumour cells is not always predictive of long term disease-free survival. More importantly, PBSC collections from AML patients are not contaminated by leukaemic cells if the BM is disease-free. PMID- 9877269 TI - Prolonged isolated thrombocytopenia after hematopoietic stem cell transplantation: morphologic correlation. AB - Prolonged isolated thrombocytopenia, defined as recovery of other cell counts with continuous dependence on platelet transfusions for greater than 90 days after hematopoietic stem cell transplantation (HSCT), develops in approximately 5% of patients who undergo HSCT. Although the clinical conditions associated with prolonged isolated thrombocytopenia have been studied, a systematic review of bone marrow biopsies has not been performed and the pathophysiologic basis has not been defined. We reviewed all HSCT at one center from 1990 to 1995 (n = 454) and found 12 cases that met criteria for prolonged isolated thrombocytopenia (incidence = 12/454 or 3%). Bone marrow core biopsies from 12 patients with prolonged isolated thrombocytopenia were reviewed to determine cellularity, numbers of megakaryocytes, the presence of atypical forms, and clusters of megakaryocytes. These marrow megakaryocyte counts were compared to age and disease matched controls, and 11 normal donors. Patients (aged 1-56 years, mean 32 years) who underwent HSCT (four sibling HLA-identical, five autologous bone marrow, three autologous peripheral stem cell) with prolonged isolated thrombocytopenia had a statistically significant lower absolute megakaryocyte count in bone marrow biopsies performed before transplantation and more than 30 days after transplantation compared to control patients (aged 4 months to 50 years, mean 31 years) who underwent HSCT (four sibling HLA-identical, four autologous bone marrow, four autologous peripheral stem cell) for similar conditions. No apparent differences were seen in size of megakaryocytes, nuclear cytoplasmic ratios, or clustering of megakaryocytes. Overall marrow cellularities were similar in the three groups. These findings suggest that decreased differentiation of megakaryocytes from stem cells, rather than ineffective platelet production or peripheral destruction of platelets, causes prolonged isolated thrombocytopenia in HSCT patients. Low megakaryocyte counts prior to HSCT may be a useful prognostic indicator, as this feature was associated with the development of prolonged isolated thrombocytopenia. PMID- 9877270 TI - Adhesion molecules on peripheral blood-derived CD34+ cells: effects of cryopreservation and short-term ex vivo incubation with serum and cytokines. AB - The homing of hematopoietic precursor cells (HPC) within the bone marrow is most likely to be mediated by specific adhesion via surface receptors to cellular and extracellular matrix (ECM) components and to be regulated by cytokines. We investigated the effects of serum and cytokines on the expression of adhesion molecules on cryopreserved and fresh peripheral blood-derived progenitor cells (PBPC) and on the adhesion of PBPC to various ECM proteins. PBPC were collected from patients by leukapheresis during G-CSF-supported recovery from conventional cancer chemotherapy. Freezing markedly reduced the fraction of CD34+ cells with L selectin (CD62L) expression from 62 to 11% and also diminished the fluorescence intensity for the integrin subunits CD29 and CD49d on CD34+ cells. A 14 h incubation of thawed PBPC with serum induced re-expression of adhesion molecules. The addition of the cytokine cocktails (G-CSF + SCF + IL-3 + IL-11 or IL-4 + IL 1beta + IFN-gamma) or MGDF, however, exerted no effects in addition to serum alone. Furthermore, when compared to serum alone, the addition of cytokine cocktails or MGDF did not alter the fraction of fresh PB-CD34+ cells adhering to collagen I, collagen IV, fibronectin, laminin or vitronectin. HPC adhesion to ECM components might be refractory to short-term alterations of the cytokine environment. Alternatively, longer incubation times or other cytokines may be necessary to modulate the expression of adhesion molecules on hematopoietic progenitor cells or adhesion itself under ex vivo conditions. PMID- 9877271 TI - Analysis of rhG-CSF-effects on platelets by in vitro bleeding test and transcranial Doppler ultrasound examination. AB - Experimental evidence suggests a stimulatory effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on both platelets and coagulation. RhG-CSF is increasingly used to stimulate healthy volunteer donors for blood stem cell mobilization. We therefore assessed 25 healthy donors receiving rhG-CSF for changes in in vitro bleeding test (IVBT), coagulation parameters and cerebral microembolism by transcranial Doppler (TCD) ultrasound. A significant shortening of IVBT was found on day 4 of rhG-CSF administration together with increased levels of fibrinogen and factor VIII and reduced activities of protein C and protein S. Although these changes are quite small it is possible that they may lead to a hypercoagulable state especially in donors with other risk factors for thromboembolism. However, TCD examination failed to detect any signs of microembolism. We therefore conclude that rhG-CSF leads to significant changes in coagulation parameters, but has no effect on TCD detectable microembolism as a stroke risk factor. However donors receiving rhG CSF should be examined carefully to detect pre-existing changes in the coagulation system and we would like to suggest a routine thrombophilia screen. PMID- 9877272 TI - Successful cryopreservation of purified autologous CD34+ cells: influence of freezing parameters on cell recovery and engraftment. AB - Conventional hematopoietic stem cell cryopreservation methods use a DMSO concentration of 10%. However, cells manipulated ex vivo may require more refined freezing protocols adapted to the specific cell suspension. In this retrospective study, we evaluated the results obtained with CD34+ cells purified from peripheral blood of 39 patients on the CEPRATE SC System and frozen in 7.5% DMSO with a view to transplantation. The post-freezing recovery of progenitor cells was 89.4 +/- 27.87% for CD34+ cells, 59.13 +/- 36.93% for CFU-GM, and 53.49 +/- 40.71 for BFU-E. Neither the purity of the suspension nor the nucleated cell density during freezing was predictive of cell recovery. No difference was observed between cells stored in vials and bags. Thirty-seven patients transplanted with the concentrated CD34+ fraction received 4.46 x 10(6) CD34+ cells/kg and 33.04 x 10(4) CFU-GM/kg. The median time to granulocyte (>0.5 x 10(9)/l) and platelet (>50 x 10(9)/l) engraftment was 11 and 13 days, respectively. Only cell density and the infused number of CD34+ cells and CFU-GM were significantly related to hematological recovery. Our data suggest that purified CD34+ cells can be successfully cryopreserved in 7.5% DMSO and may represent a first step in establishing freezing parameters for selected CD34+ cells. PMID- 9877273 TI - Differential effects of cyclosporin A on the alloreactivity of fresh and ex vivo expanded T lymphocytes. AB - GVHD remains a major source of morbidity and mortality after non-T cell-depleted BMT. The use of donor T lymphocytes expressing a suicide gene could lead to specific immunomodulation after BMT. We are currently evaluating such an approach in a phase I clinical study. A 12-day ex vivo expansion is required to generate gene-modified donor T lymphocytes. CsA is commonly used for GVHD prophylaxis. We analyzed, in a murine GVHD model, the effects of CsA administration on the alloreactivity of fresh or ex vivo-expanded T cells. Variable amounts of fresh or ex vivo-expanded T cells were administered in conjunction with a marrow graft to lethally irradiated allogeneic mice. As expected, a protective effect of CsA with a delayed GVHD-related mortality (P < 0.01 vs saline treatment) was observed in mice receiving fresh splenocytes. However, CsA treatment had no effect (P = NS) in mice experiencing lethal GVHD induced by ex vivo-expanded T cells whether or not the T cells had been 'rested' in low-dose IL-2 prior to in vivo administration. In agreement with the in vivo findings, CsA also inhibited the in vitro proliferation of alloreactive fresh T cells while having no significant inhibitory effect on the alloreactivity of ex vivo-expanded T lymphocytes. Overall, we demonstrate that the alloreactivity of ex vivo-expanded T lymphocytes is not sensitive to CsA and that this differential effect of CsA is not related to the alloreactive potential of the infused T cells. These findings could be highly relevant when considering allogeneic T cell therapy approaches. PMID- 9877274 TI - Changes in L-selectin expression on CD34-positive cells upon cryopreservation of peripheral blood stem cell transplants. AB - Several studies have pointed out that L-selectin on CD34-positive cells plays a role in haematopoietic reconstitution after peripheral blood stem cell (PBSC) transplantation. Since it is known that a decrease in L-selectin expression in lymphocytes and granulocytes can be induced by a variety of stress situations, we have investigated in this study whether the freeze-thawing procedure, used in PBSC transplantation, would affect L-selectin expression on CD34+ stem cells. Flow cytometry was performed by labelling the cells with anti-CD34 (HPCA2 PE) and anti-CD62L (FMC46 FITC). The leucapheresis procedure itself caused a slight decrease of L-selectin expression on CD34 cells in 11 out of 12 cases (mean decrease of the percentage of positive cells 11.9; range 6-23). A much larger decrease was found upon freeze-thawing: a mean of 39% (range 4-78% in 27 cases) compared to fresh material. To determine if L-selectin expression might be up regulated after cryopreservation, thawed transplant samples (n = 11) were incubated at 37 degrees C in RPMI with 10% FCS at 5% CO2. Already early in the course of incubation two CD34-positive populations appeared in the blast region, characterized by either a low or high forward scatter. Simultaneous viability staining with the DNA dye 7-Amino Actinomycin D and the DNA/RNA dye Syto16 revealed that the population with low forward scatter was apoptotic while the population with the high forward scatter was non-apoptotic. The latter population is considered to be most relevant for transplantation. In this population an increase of L-selectin expression after overnight incubation was observed in 8/11 samples up to values of 46-120% of the values of the fresh nonfrozen samples. In addition, the mean fluorescence intensity was significantly increased in 10/11 cases. Kinetic experiments with shorter incubation times revealed that only part of the leucapheresis samples (two out of 8) showed an increase of L-selectin expression within 4 h. In addition, a decrease of L-selectin expression was found upon CD34 purification from fresh leucapheresis material by magnetic isolation (decrease ranging from 59 to 92%, n = 5). In contrast to frozen samples, L selectin reappearance was seen already within 4 h of incubation in all samples. Both the loss of L-selectin expression on CD34 cells occurring upon freeze thawing, the emergence of apoptosis, as well as the recovery of L-selectin expression on non-apoptotic cells varies largely between individual leucapheresis samples, and therefore it is concluded that such processes should be considered when correlations with clinical outcome after transplantation are made. PMID- 9877275 TI - Bone marrow transplantation as treatment for X-linked immunodeficiency with hyper IgM. AB - We report a 10-year-old boy with a severe form of immunodeficiency with hyper-IgM who underwent successful bone marrow transplantation with his HLA-matched sister as donor. Busulfan (20 mg/kg) and cyclophosphamide (200 mg/kg) were used as conditioning. The post-transplant course was uneventful. He is alive 25 months later with full hematological and immunological reconstitution. PMID- 9877276 TI - Acute GVHD involving the gastrointestinal tract and infestation with Blastocystis hominis in a patient with chronic myeloid leukaemia following allogeneic bone marrow transplantation. AB - A 21-year-old female underwent allogeneic bone marrow transplantation (ABMT) from her HLA matched brother for chronic myeloid leukaemia in the chronic phase. Four weeks post transplant she developed tenesmus, mucoid stool mixed with blood and lower abdominal pain. This rapidly progressed to greenish watery diarrhoea with flakes of mucous membrane floating in it, conforming to the classical clinical description of acute GVHD of the bowel. Stool microscopy showed profuse numbers of Blastocystis hominis, a parasite with doubtful pathogenicity in an immunocompetent host. In the present case the parasite played a pathogenic role as evidenced by the profuse number in the stool sample, focal neutrophil infiltration of the rectal mucosa, initial presentation of the patient with dysentery, and requirement for prolonged metronidazole therapy to eradicate the parasite and cure the diarrhoea. She also had grade I GVHD of the liver and skin. In developing tropical countries, hitherto unreported parasitic infestations may complicate the picture of acute GVHD. PMID- 9877277 TI - Improved peripheral nerve conduction, EEG and verbal IQ after bone marrow transplantation for adult metachromatic leukodystrophy. AB - A 28-year-old woman with a 4 year history of slowly progressing 'frontal dementia' was diagnosed as having adult metachromatic leukodystrophy and was followed for 4 years after bone marrow transplantation (BMT). MRI, neurophysiological tests (EEG, ENeG, VEP, SEP and BAEP) and neuropsychological assessment were performed before, and repeatedly after BMT. MRI showed symmetrical white matter lesions in the frontal and parieto-occipital lobes and in the corpus callosum. EEG showed frontal and temporal slow wave abnormalities and nerve conduction was slow. Neuropsychological tests showed cognitive impairment in executive functions, decline in visuospatial-constructive and spatial memory tasks and disorganized thinking. IQ was low (52), with slightly better values for verbal IQ than for performance IQ. After BMT, the patient was followed for 4 years. Clear improvements were seen in EEG, in peripheral nerve conduction and in neuropsychological tests (especially in verbal IQ). MRI findings were unchanged. We believe that the improvement in our patient resulted from the bone marrow transplantation. PMID- 9877278 TI - Noninvasive mechanical ventilation in a patient with respiratory failure after hematopoietic progenitor transplantation. AB - Respiratory failure requiring orotracheal intubation (OTI) and mechanical ventilation (MV) is almost always a fatal complication in patients who undergo hematopoietic progenitor transplantation (HPT). We present the case of a woman who suffered respiratory failure with bilateral infiltrates on a chest X-ray taken on day +14 following autologous bone marrow transplantation. We managed the patient satisfactorily with noninvasive ventilation, avoiding OTI. We believe that patients with non-progressive pulmonary lesions and without multiple system organ failure, may be correctly managed with noninvasive positive-pressure ventilation (NPPV). PMID- 9877279 TI - Bone marrow necrosis in bone marrow transplantation: the role of MR imaging. AB - We describe an ALL patient who developed extensive bone marrow necrosis at the time of relapse 2 months after allogeneic bone marrow transplantation from an HLA identical sibling. The excruciating and diffuse bone pain, fever and precipitous drop in peripheral blood counts were characteristic. This case illustrates the importance of repeat bone marrow biopsies for the diagnosis of disease relapse and the potential application of MR imaging in the assessment of patients with bone marrow necrosis. PMID- 9877280 TI - Inhibition of tumor necrosis factor and subsequent endotoxin shock by pirfenidone. AB - Tumor necrosis factor-alpha (TNF) is an extremely potent cytokine which is involved in the pathogenesis of a number of diseases. Interruption of its synthesis can result in a reduction of inflammation and subsequent pathology. A new experimental drug pirfenidone (5-methyl-L-phenyl-2-(1H)-pyridone, trade name: Deskar) has been reported to have beneficial effects for the treatment of certain fibrotic diseases. The present study describes the inhibition of TNF in vitro as well as the inhibition of circulating TNF in vivo by pirfenidone. Isolated, thioglycollate-induced peritoneal macrophages (Mphi) from C57BL/6 mice were exposed to either lipopolysaccharide (LPS) or mannosylated bovine serum albumin then incubated with 0.1-0.9 mg/ml of pirfenidone. This substance inhibited the production of TNF in a dose-dependent manner as measured by ELISA. One i.p. injection of either 100 or 200 mg/kg pirfenidone inhibited the induction of circulating TNF following a single i.v. injection of LPS. Endotoxin shock was induced in mice using an i.p. injection of galactosamine and LPS. The higher dose of pirfenidone (200 mg/kg) completely inhibited shock and subsequent mortality. Lower doses of pirfenidone or administration either prior to or post challenge only partially inhibited symptoms. These results indicate that pirfenidone is able to inhibit both TNF induction and subsequent endotoxin shock. Additional studies are warranted to establish this drug as a potential treatment for diseases where TNF plays a major role. PMID- 9877281 TI - Low levels of ionic mercury modulate protein tyrosine phosphorylation in lymphocytes. AB - The ability of ionic mercury to induce protein tyrosine phosphorylation in mouse spleen cells and in the mouse WEHI-231 B-cell lymphoma was investigated. We have confirmed previous studies which showed that exposure to high levels (several hundred microM) of mercury lead to very large increases in the level of protein tyrosine phosphorylation in these cell systems. However we have also demonstrated that low levels (in the order of 0.1 to 1.0 microM) of mercury also significantly upregulate protein tyrosine phosphorylation. Mercury induced protein tyrosine phosphorylation is inhibited by the mercury chelator penicillamine and by pretreating treating target cells with the sulfhydryl blocking reagent N hydroxymaleimide. These results suggest that exposure to low levels of mercury could potentially interfere with lymphocyte signal transduction and so offer a possible explanation as to how mercury exposure could lead to immune cellular dysfunction. On a molecular level, the results suggest that the site(s) of action with respect to mercury dependent induction of protein tyrosine phosphorylation is likely a free disulfide group or groups located on the outer leaflet of the plasma membrane. PMID- 9877282 TI - Hydroxyl radical scavengers inhibit TNF-alpha production in mononuclear cells but not in polymorphonuclear leukocytes. AB - The hydroxyl radical (HO*) scavengers dimethylthiourea (DMTU), tetramethylthiourea (TMTU), dimethylsulfoxide (DMSO) and deferoxamine (DFX), the latter being an iron chelator which prevents HO* formation by blocking the Fenton reaction, were found to inhibit TNF-alpha production in LPS-stimulated human PBMC but not in PMN. Furthermore, this effect was not LPS-specific, as TNF-alpha production was reduced by HO* radical scavengers to a similar extent upon stimulation of PBMC with immune complexes (IC), concanavalin A (Con A) and phorbol myristate acetate (PMA). Other scavengers such as glutathione (GSH), N acetylcysteine (NAC), ascorbic acid (ASC) and mannitol (MAN) do not have effect on the production of TNF-alpha either in PBMC or PMN. These results provide evidence that the participation of ROI in the regulation of TNF-alpha production differ in different cell types. Particularly, the data presented in this work indicate that HO* radicals have a central role in the production of this inflammatory cytokine by human PBMC. PMID- 9877283 TI - Comparative study on effects of pentoxifylline, prednisolone and colchicine in experimental alveolitis. AB - Neutrophil alveolitis is a hallmark of cryptogenic fibrosing alveolitis (CFA), known for its poor prognosis. Corticosteroids, as the remedy of choice, are ineffective in a majority of patients. More and more evidence indicates that pentoxifylline (POF) could be an effective therapeutic alternative. Furthermore, colchicine has been proposed for therapy of CFA for many years now. We conducted an experimental study comparing the efficacy of these drugs in preventing neutrophil alveolitis in vivo. Alveolitis was induced in male rats by intratracheal instillation of bleomycin. Treatment consisted of daily injections of POF i.p., colchicine i.p., or prednisolone i.m. After 8 days the animals were sacrificed and body weights, cell differentials in BAL, amount of proliferating interstitial cells as determined by KI-67 staining of lung tissue, and collagen concentrations in lungs were determined. Bleomycin instillation was followed by a significant weight loss in the animals, a neutrophil alveolitis in BAL and an increased amount of proliferating cells in lung interstitium. POF significantly inhibited any of the parameters named, whereas prednisolone and colchicine had little effect. Data cannot be applied directly in human disease. There are however many similarities between CFA and bleomycin-induced lung injury and alveolitis. We conclude that POF is an effective inhibitor of neutrophil alveolitis, whereas neither colchicine nor prednisolone exerted significant influence in our model. We suggest POF effects should be further investigated regarding anti-inflammatory and anti-fibrotic properties. PMID- 9877284 TI - Interactive effects of cocaine and gender on thymocytes: a study of in vivo repeated cocaine exposure. AB - This study used a mouse model including both sexes to assess the impact of repeat cocaine exposure on the differentiation and function of T cell in thymus. Cocaine hydrochloride in 0.9% saline, 5 mg or 40 mg/kg, was administrated by i.p. injection to C57BL/6 mice for 10 days. Thymocytes were obtained 24 h after the 10th injection. Repeat in vivo cocaine exposure inhibited the proliferation of T lymphocytes in response to Con-A and Con-A plus anti-CD28. The proliferation induced by IL-2 in the Con-A stimulated T blasts was attenuated in cocaine treated mice. These effects were seen at a lower cocaine dose in female mice. The total number of thymocytes was reduced. Although the percentage of mature thymocytes (CD4+ CD8- and CD4- CD8+ cells) was not altered, the absolute cell numbers were attenuated. Both percentage and absolute cell number of immature thymocytes (CD4+ CD8+) decreased and the pre-mature (CD4- CD8-) cells increased. CD28 and CD25 expression were attenuated in Con-A stimulated thymocytes of mice treated with cocaine at 40 mg/kg. Interleukin 2 production was not significantly altered, however, gamma-IFN production was decreased by cocaine exposure at 40 mg/kg. In conclusion, cocaine exerts inhibitory effects on the function of mature thymocytes, and on the differentiation of thymocytes. A gender difference in response to cocaine was noted in that female mice were more sensitive to lower dose of cocaine exposure. PMID- 9877285 TI - Effect of d-fenfluramine on the lymphocyte response of HIV+ humans. AB - The objective of this study was to analyse the effect of d-dexfenfluramine (d FEN) on the human lymphocyte response, in vitro. Experiments were designed to determine whether d-FEN augments specific human immune parameters associated with protection from opportunistic microbial pathogens and particularly focuses on d FEN as a means by which to augment the function of CD8+ and CD4+ lymphocytes. Lymphocytes were examined for three reasons: (1) for their ability to inhibit the growth of Candida albicans; (2) for their ability to proliferate in response to a mitogen; and, (3) their cytokine profile (vis., production of IL-2, IFN-gamma and TNF-alpha). Peripheral blood mononuclear cells (PBMC) were obtained from 20 HIV+ patients. The patients were diagnosed as HIV+ within the past 0.5-9 years. d-FEN was found to augment the capacity of CD8+ lymphocytes to inhibit the growth of the opportunistic microbial pathogen, C. albicans. d-FEN enhanced the capacity of CD4+ lymphocytes to proliferate in response to the mitogen, Concanavalin A, and to increase the amount of IL-2 produced by CD4+ and CD8+ lymphocytes from AIDS patients. d-FEN increased the number of CD4+ and CD8+ lymphocytes that produced IFN-gamma from either non-AIDS or AIDS patients and increased the number of AIDS patient's CD8+ lymphocytes that produce TNF-alpha. These in vitro data suggest that d-FEN may be effective in enhancing immune function in immunocompromised individuals. PMID- 9877286 TI - Fosfomycin, an antibiotic, possessed TGF-beta-like immunoregulatory activities. AB - The regulatory effects of fosfomycin (FOM) were correlated closely with the multifunction of TGF-beta in the modulation of immune responses in vivo and in vitro. LPS-induced polyclonal IgM and IgG antibody responses were depressed at 3 days after the initial culture and subsequently enhanced at day 10 by FOM or TGF beta. Neither FOM nor TGF-beta inhibited LPS-induced IgA antibody responses, whereas dexamethasone (DX) reduced polyclonal IgM, IgG and IgA antibody responses wholly. The suppression of antibody responses and Mv1Lu cell proliferation induced by FOM or TGF-beta was partly overcome with soluble TFG-beta receptors (sRIII). Oral, i.v. and i.p. administration of FOM exhibited similar enhanced SRBC-specific antibody responses to that seen after oral administration of TGF beta. The addition of FOM and latent TGF-beta inhibited the proliferation of Mv1Lu cells, but FOM did not lead to an increase in plasmin activities, which convert latent to active TGF-beta, and further the expression of TGF-beta receptors on the cell surface. In addition, FOM failed to enhance TGF-beta secretion. These findings suggest that immunomodulation of FOM results in increased sensitivity of cells to TGF-beta. PMID- 9877287 TI - Effect of mild chronic stress, as a model of depression, on the immunoreactivity of C57BL/6 mice. AB - Numerous studies correlate the state of depression with some abnormalities in the immune response, such as increased numbers of white blood-cells, alterations in sub-populations of leucocytes, suppression of cytotoxic activity of natural killer cells, increased levels of some autoantibodies and acute-phase proteins. Some of these changes have been attributed to autoimmunological reactions. While the possibilities to evaluate some reactions in depressed patients are limited, an animal model of depression could well simulate this clinical situation, and the chronic mild state of stress is a well accepted one. After undergoing stress for three-weeks, C57BL/6 mice demonstrate in the present study a decrease in thymus weight, as well as increased interleukin-1 and decreased interleukin-2 production. Splenocytes of the depressed mice exert a decrease in natural-killer cell activity, in the proliferative response to Concanavalin-A, interleukin-1 and anti-CD3 monoclonal antibodies and an increase in the proliferative response to lipopolysaccharides and pokeweed mitogens. Our results also suggest that chronic stress-induced activation of suppressor cells in the spleen, due to elimination of CD8+ cells, increase the proliferation of splenocytes in response to mitogens of T cells. PMID- 9877288 TI - Computer-generated three-dimensional reconstructions of serially sectioned mouse embryos. AB - We have been involved with a group of computer scientists and anatomists in the development of computer-based methodologies that not only combine the advantages of scanning electron microscopy and conventional histology, but provide the additional dimension of tissue recognition. The latter is achieved by the appropriate labelling of tissues and structures by delineation or 'painting'. Individually segmented anatomically defined tissues can be highlighted in a particular colour and viewed either in isolation or in combination with other appropriately labelled tissues and organs. Tissues can be shown in any orientation either as a transparent overlay on computer-generated histological sections or as 3-D images without the histological background. An additional feature of the system is that computer graphics technology combined with 3-D glasses now also allows the viewer to see the object under analysis in stereo. This facility has been found to be particularly helpful in drawing attention to topological relationships that had not previously been readily noted. As the mouse is now the mammalian model of choice in many areas of developmental research, it is of critical importance that a basic level of skill is available in the research community in the interpretation of serially sectioned material, for example, for the rapidly expanding field in which gene expression studies play a significant role. It is equally important that there is an understanding of the dynamic changes that occur in relation to the differentiation of the various organ systems seen in these early stages of development. What we emphasise here is the additional information that it is possible to gain from the use of this tool which, in our view, could not readily have been gained from the analysis of scanning electron micrographs or by studying conventional serial histological sections of similar stages of mouse embryonic development. The methodology has been developed as part of a large project to prepare a database of mouse developmental anatomy covering all stages from fertilisation to birth in order to allow the accurate spatial mapping of gene expression and cell lineage data onto the digital Atlas of normal mouse development. In this paper we show how this digital anatomical Atlas also represents a valuable teaching aid and research tool in anatomy. PMID- 9877289 TI - Computer-aided interactive three-dimensional reconstruction of the embryonic human heart. AB - Despite the fact that development of the human embryo heart is of considerable clinical importance, there is still disagreement over the process and the timing of events. It is likely that some of the conflicting accounts may have arisen from difficulties in describing and visualising 3-dimensional structures from 2 dimensional sections. To help overcome this problem and to improve our understanding of the development of the heart, we have devised techniques for the production of interactive 3D models reconstructed from serial histological sections of human embryos. Our method uses commercial software designed for the creation of 3D models and virtual reality environments. The ability to construct interactive visual images which both illustrate and communicate complex 3D information contributes to our understanding of the complex developmental changes occurring in embryogenesis. PMID- 9877290 TI - Algorithms for radiological image registration and their clinical application. AB - This paper reviews recent work in radiological image registration and provides a classification of image registration by type of transformation and by methods employed to compute the transformation. The former includes transformation of 2D images to 2D images of the same individual, transformation of 3D images to 3D images of the same individual, transformation of images to an atlas or model, transformation of images acquired from a number of individuals, transformations for image guided interventions including 2D to 3D registration and finally tissue deformation in image guided interventions. Recent work on computing transformations for registration using corresponding landmark based registration, surface based registration and voxel similarity measures, including entropy based measures, are reviewed and compared. Recently fully automated algorithms based on voxel similarity measures and, in particular, mutual information have been shown to be accurate and robust at registering images of the head when the rigid body assumption is valid. Two approaches to modelling soft tissue deformation for applications in image guided interventions are described. Validation of complex processing tasks such as image registration is vital if these algorithms are to be used in clinical practice. Three alternative validation strategies are presented. These methods are finding application outside the original domain of radiological imaging. PMID- 9877291 TI - Advances in three-dimensional diagnostic radiology. AB - The maturity of current 3D rendering software in combination with recent developments in computer vision techniques enable an exciting range of applications for the visualisation, measurement and interactive manipulation of volumetric data, relevant both for diagnostic imaging and for anatomy. This paper reviews recent work in this area from the Image Sciences Institute at Utrecht University. The processes that yield a useful visual presentation are sequential. After acquisition and before any visualisation, an essential step is to prepare the data properly: this field is known as 'image processing' or 'computer vision' in analogy with the processing in human vision. Examples will be discussed of modern image enhancement and denoising techniques, and the complex process of automatically finding the objects or regions of interest, i.e. segmentation. One of the newer and promising methodologies for image analysis is based on a mathematical analysis of the human (cortical) visual processing: multiscale image analysis. After preprocessing the 3D rendering can be acquired by simulating the 'ray casting' in the computer. New possibilities are presented, such as the integrated visualisation in one image of (accurately registered) datasets of the same patient acquired in different modality scanners. Other examples include colour coding of functional data such as SPECT brain perfusion or functional magnetic resonance (MR) data and even metric data such as skull thickness on the rendered 3D anatomy from MR or computed tomography (CT). Optimal use and perception of 3D visualisation in radiology requires fast display and truly interactive manipulation facilities. Modern and increasingly cheaper workstations ( < $10000) allow this to be a reality. It is now possible to manipulate 3D images of 256 at 15 frames per second interactively, placing virtual reality within reach. The possibilities of modern workstations become increasingly more sophisticated and versatile. Examples presented include the automatic detection of the optimal viewing angle of the neck of aneurysms and the simulation of the design and placement procedure of intra-abdominal aortic stents. Such developments, together with the availability of high-resolution datasets of modern scanners and data such as from the NIH Visible Human project, have a dramatic impact on interactive 3D anatomical atlases. PMID- 9877292 TI - Correlated histological and physiological observations on a case of common sensory output and motor input of the bag1 fibre and a chain fibre in a cat tenuissimus spindle. AB - In muscle spindles of the cat, independent control of dynamic and static components of the response of the primary sensory ending to stretch is provided by separate motor inputs to the various kinds of intrafusal muscle fibre: dynamic axons (gamma or beta) to the bag1 fibres and static axons to the bag2 (typically gamma only) and chain (gamma or beta) fibres. Nonlinear summation of separately evoked effects during combined stimulation of dynamic and static motor axons appears to be due to mutual resetting by antidromic invasion of separate encoding sites, leading to partial occlusion of the momentarily lesser response by the greater. The encoding sites are thought to be located within the primary ending's preterminal branches which from first-order level are normally segregated to the bag1 fibre and to the bag2 and chain fibres. Here we describe the analysis of a special case that arose in a histophysiological study which had shown that the degree of occlusion was related to the minimum number of nodes between the putative encoding sites. Three-dimensional reconstruction of the primary ending revealed that the terminals of one chain fibre were derived entirely from the first-order branch that supplied the bag1 fibre, including one terminal that was shared directly with the bag1 (sensory cross-terminal). The other first-order branch supplied the bag2 and remaining chain fibres as normal. The degree of occlusion seen during simultaneous stimulation of a dynamic beta axon and a static gamma axon indicated that the encoding sites were separated by both first order branches. Schematic reconstruction of the motor innervation revealed that the static gamma axon was most unlikely to have supplied the chain fibre which shared sensory terminals with the bag1, but that these fibres also shared a motor input with histological characteristics of beta type. Ramp-frequency stimulation of the dynamic beta axon at constant length evoked a driving effect which persisted after fatiguing the extrafusal component and was therefore explicable on the basis of the observed pattern of motor innervation, though the identity of the axon could not be conclusively proved. Individually, instances of shared sensory terminals and motor input of bag1 and chain fibres are rare in the cat; their combination in a single spindle with correlated physiology is described here for the first time. The observation is considered in relation to the importance of dynamic and static segregation in motor control, since it may imply that there is a lower limit to the degree of segregation that the developmental programme can provide. PMID- 9877293 TI - The effect of temporary reductions in incubation temperature on growth characteristics and lipid utilisation in the chick embryo. AB - The effect(s) of finite exposure(s) to 22.0 degrees C on embryo weight gain, 3rd tarsus length, transfer of lipid from the yolk and uptake of specific lipid components by the liver, were measured in the chick embryo. Embryo growth was completely suspended over the period of temperature reduction, but resumed unimpaired when the optimum temperature of 37.5 degrees C was restored. Two successive periods of embryonic exposure to 22.0 degrees C (36 h at d 3 followed by 24 at d 10) were additive in their effect on growth retardation. Embryos subjected to temperature reductions on d 3 of incubation displayed significant changes in lipid concentrations and polyunsaturated fatty acid profiles within the liver when measured at d 18 of incubation. Yolk lipid uptake was significantly reduced but not suspended by embryonic exposure to 22.0 degrees C. It is suggested that the observed suspension of embryonic development was the result of a temperature dependent reduction in metabolism according to a Q10 approximately 2, which is typical of an ectothermic organism. PMID- 9877294 TI - In situ detection of apoptotic cells by TUNEL in the gill epithelium of the developing brown trout (Salmo trutta). AB - Apoptosis is a form of naturally occurring cell death during development and it is characterised by extensive DNA fragmentation. Apoptosis is easily detected in the gill epithelium of brown trout embryos in ultrathin sections (Rojo et al. 1997). Here we provide the first biochemical evidence for apoptosis in the gill epithelium of brown trout embryos, using in situ end-labelling of DNA breaks (Gavrieli et al. 1992). Embryos at d 57 of development as well as those at hatching, were processed to analyse the distribution of apoptotic cells in the gills. The extent of apoptosis revealed by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labelling method technique is considerably greater than apoptosis detected by nuclear morphology. This method revealed that apoptosis was frequent at hatching, although it was also present during embryonic development. The presence and distribution of stained nuclei were different depending on the developmental stage. In embryos of 57 d, apoptotic flattened nuclei were dispersed in the gill epithelium, whereas at hatching, they were mainly grouped in the tips of the filaments and in the gill arches. TUNEL also revealed a distinct pattern of nuclear staining: at hatching, the intense staining covered the entire cell, but in embryos it was restricted to the nucleus. These results show the functional relevance of apoptosis at hatching, when apoptosis seems to be the unique process by which cell numbers in the gill epithelium are adjusted, in order to prepare for the new extrinsic conditions affecting the free-living life of alevins. PMID- 9877295 TI - Thyroid transcription factor-1, hepatocyte nuclear factor-3beta and surfactant protein A and B in the developing chick lung. AB - Expression of surfactant proteins SP-A, SP-B and the transcription factors TTF-1 and HNF-3beta was identified by immunohistochemistry in the developing chicken. SP-B, a small hydrophobic peptide critical for lung function and surfactant homeostasis in mammals, was detected in the epithelial cells of parabronchi in embryonic chicken lung from the 15th day of incubation, prior to the onset of the breathing movements and was expressed at high levels in the posthatching chicken lung. SP-A, an abundant surfactant protein involved in innate defence of the mammalian lung, was detected in the chick embryo in subsets of epithelial cells in the mesobronchus, starting from d 15 and was detected in the posthatching chicken lung. The transcription factors hepatocyte nuclear factor 3beta (HNF 3beta) and thyroid transcription factor-1 (TTF-1), both regulators epithelial cell differentiation and gene expression in mammalian species, were detected at the onset of lung bud formation (d 4 of incubation) and throughout lung development. Abundant nuclear expression was detected in nuclei of respiratory epithelial cells of developing bronchial tubules for both transcription factors. In contrast to the surfactant proteins, expression of both TTF-1 and HNF-3beta decreased markedly in posthatching chicken lung. The expression of SP-A and SP-B in chick lung demonstrates the conservation of surfactant proteins in vertebrates. The temporospatial pattern of TTF-1 and HNF-3beta overlaps with that of SP-A and SP-B, supporting their potential roles in chick lung development and demonstrating the conservation of regulatory mechanisms contributing to gene expression in respiratory epithelial cells in vertebrates. PMID- 9877296 TI - Angioarchitecture of the coeliac sympathetic ganglion complex in the common tree shrew (Tupaia glis). AB - The angioarchitecture of the coeliac sympathetic ganglion complex (CGC) of the common tree shrew (Tupaia glis) was studied by the vascular corrosion cast technique in conjunction with scanning electron microscopy. The CGC of the tree shrew was found to be a highly vascularised organ. It normally received arterial blood supply from branches of the inferior phrenic, superior suprarenal and inferior suprarenal arteries and of the abdominal aorta. In some animals, its blood supply was also derived from branches of the middle suprarenal arteries, coeliac artery, superior mesenteric artery and lumbar arteries. These arteries penetrated the ganglion at variable points and in slightly different patterns. They gave off peripheral branches to form a subcapsular capillary plexus while their main trunks traversed deeply into the inner part before branching into the densely packed intraganglionic capillary networks. The capillaries merged to form venules before draining into collecting veins at the peripheral region of the ganglion complex. Finally, the veins coursed to the dorsal aspect of the ganglion to drain into the renal and inferior phrenic veins and the inferior vena cava. The capillaries on the coeliac ganglion complex do not possess fenestrations. PMID- 9877297 TI - Supraspinal connections and termination patterns of the parabrachial complex determined by the biocytin anterograde tract-tracing technique in the rat. AB - We have re-evaluated, using the anterograde tracer biocytin, supraspinal efferent projections from the parabrachial complex (PBN) to gain new information about the nature of its connections and nerve terminal patterns. We selectively injected biocytin into the 3 main regions of the nucleus (lateral PBN, medial PBN and Kolliker-Fuse nucleus). We observed distinct groups of ascending and descending fibres of different calibre from the PBN running throughout the brain and reaching many brain areas involved in the regulation of autonomic function. Here we detected labelled bouton-like terminals and fibres with en-passage varicosities. The ascending efferents from the lateral PBN mainly reached the reticular, raphe and thalamic nuclei, the zona incerta (ZI), central nucleus of the amygdala (CeA) and lateral area of the periaqueductal grey (PAG). Thin descending efferents reached the ventral region of the solitary tract nucleus (STN). The ascending efferents from the medial PBN were seen in the raphe nuclei, reticular nuclei, ventral and lateral areas of the PAG, thalamic nuclei, and in the medial and lateral nuclei of the amygdala. Descending efferents were seen in the STN and in some reticular nuclei. The ascending projections from the Kolliker Fuse targeted the ventral area of PAG, CeA, ZI, lateral hypothalamic area, ventromedial thalamic nucleus and, with only a few terminals, the ipsi and contralateral reticular area. A large number of descending efferents reached STN, caudal and paragigantocellular reticular nuclei. The higher sensitivity of biocytin compared with other types of markers allowed us to determine more effectively the distribution, nature and extent of the supraspinal PBN connections. This suggested that in several nerve circuits the PBN probably plays a more important role than previously thought. PMID- 9877298 TI - Pathological bone changes in the mandibles of wild red deer (Cervus elaphus L.) exposed to high environmental levels of fluoride. AB - A macroscopic, microscopic and scanning electron microscope study was performed on the pathological bone changes of the mandibles of wild red deer (n = 61) exhibiting severe dental fluorosis. The animals originated from a highly fluoride polluted area in Central Europe (Ore mountains and their southern foreland, Czech German border region) and constituted 11.2 % of the studied red deer sample (n = 545) from this area. Pathologically increased wear and fracture of fluorosed teeth caused a variety of mandibular bone alterations, including periodontal breakdown, periostitis, osteitis and chronic osteomyelitis. As a further consequence of severe dental attrition, opening of the pulp chamber and formation of periapical abscesses were occasionally observed. In case of severe periodontal breakdown, loss of teeth from the mandibles was found. In addition to the inflammatory bone changes, the occurrence of osteofluorotic alterations was also diagnosed in the specimens with the highest bone fluoride concentrations (> 4000 mg F-/kg dry wt). These changes comprised extended apposition of periosteal bone onto the mandibular cortex as well as deformation of the mandibular body, which was attributed to a fluoride-induced osteomalacia. The present study provided circumstantial evidence that, in addition to fluoride induced dental lesions, the occurrence of marked periodontal disease and tooth loss is an important factor responsible for a reduction of life expectancy in severely fluorotic wild red deer. PMID- 9877299 TI - The proliferative status of haematopoietic progenitor cells in the developing murine liver and adult bone marrow. AB - A class of primitive progenitor cells with high proliferative potential in vitro (HPP-CFC), has been identified in fetal liver and adult bone marrow both in murine and human systems. The kinetic properties of HPP-CFC2 and the more mature granulocyte-macrophage colony forming units (CFU-GM) derived from murine fetal liver on d13, d15 and d19 of gestation, newborn liver and neonatal liver on d3 and d8 postpartum have been evaluated and compared with the kinetic properties of these progenitor cell populations derived from adult bone marrow. The frequency of HPP-CFC2 in fetal liver was found to be greatest on d15 of gestation then subsequently declined in newborn and neonatal liver. Similarly, the highest proportion of HPP-CFC2 engaged in DNA synthesis (53+/-3%) was detected in d15 fetal liver. This proportion decreased to 13+/-2% in the liver 1 wk after birth, which is comparable to the number of HPP-CFC2 derived from adult BM which were in S-phase (10+/-1%). Production of CFU-GM was found to be greater in adult bone marrow than in either fetal or newborn liver. While the proportion of CFU-GM in S phase was high all 3 tissue samples, the greatest proportion of cycling CFU-GM (50+/-2%) was detected in d15 fetal liver. These results suggest that HPP-CFC2 derived from fetal liver are actively cycling while HPP-CFC2 derived from adult bone marrow are relatively quiescent. In contrast, a high proportion of CFU-GM derived from fetal, newborn liver and adult bone marrow are engaged in DNA synthesis. PMID- 9877300 TI - Determination of a standard site for the measurement of bone mineral density of the human calcaneus. AB - Ultrasound of the calcaneus may be used as a cheap, ionising radiation-free and easy to use indicator of skeletal status, and hence of osteoporotic fracture risk. At present ultrasound is not widely used as it suffers from high precision errors. As ultrasound parameters are determined in part by bone mineral density (BMD), an increase in the accuracy and precision of BMD measurements should reduce the precision error associated with ultrasound measurements. The aim of this study was to define an anatomical site on the calcaneus at which accurate and precise measurements of BMD can be made. Ten dry calcanei and 10 cadaveric feet were scanned using a DXA scanner; 9 anatomically defined regions (1 cm2) were selected in the posterior part of the calcaneus for analysis. The centre of region 1 was positioned halfway along the line joining the anterior border of the calcaneal tubercle and the peak of the posterior superior tubercle, and the remaining 8 regions were placed around this central area. The BMD in these 9 regions was compared with the whole bone BMD and the variability of BMD within each of the 9 regions was measured. The reproducibility of the technique was assessed by taking 10 repeated measurements of 2 bone and 2 cadaveric specimens, each specimen being removed and repositioned between measurements. Region 1 was found to be the most representative of total BMD in cadaveric feet. This region also showed the least variability of BMD and consistently gave the lowest coefficients of variation in the reproducibility study both in the bone and the cadaveric specimens. This region is hence the most suitable site on the calcaneus for measuring absolute values of and changes in BMD. The surface position of region 1 was found to be consistently 5/9 along the line at 45 degrees to the vertical, from the lateral malleolus to the heel. The identification of the surface location of region 1 relative to anatomical landmarks of the foot has enabled the same anatomical site to be measured in all subjects. This allows meaningful intersubject comparisons to be made. Preliminary data suggest that precision errors using ultrasound are also reduced when measurements are taken at this region of the calcaneus. The reduction in the precision error of ultrasound assessment of skeletal status may provide a cheap and safe way to identify individuals at risk from osteoporotic fracture. PMID- 9877301 TI - Epithelial-connective tissue boundary in the oral part of the human soft palate. AB - The papillary layer of the oral part of the human soft palate was studied in 31 subjects of different age by means of histological, immunohistochemical and scanning electron microscopical methods. For scanning electron microscopy a new maceration method was introduced. Results determine epithelial thickness, height and density of connective tissue papillae and their 3-dimensional architecture inside the lining epithelium as well as the collagenous arrangement of the openings of the glandular ducts. The individual connective tissue papillae of the soft palate are compared with the connective tissue boundary on the other side of the oral cavity. The connective tissue plateaux carrying a variable number of connective tissue papillae were found to be the basic structural units of the papillary body. The function of the epithelial-connective tissue interface and the extracellular matrix arrangement in the lamina propria are discussed in order to promote the comparability of normal with pathologically altered human soft palates. PMID- 9877303 TI - The bulbocerebellar circumolivary bundle. PMID- 9877305 TI - Characterization of amorphous ursodeoxycholic acid prepared by spray-drying. AB - The objectives of this study were to characterize the amorphous state of ursodeoxycholic acid (UDCA) samples prepared by spray-drying, and to demonstrate the applicability of thermal and water-vapour-adsorption techniques for studying the material. Amorphous UDCA was prepared by spray-drying a solution of the compound in a mixture of ethanol and dichloromethane. The amorphous material was characterized by powder X-ray diffraction, infrared (IR) spectroscopy, isothermal microcalorimetry, differential scanning calorimetry (DSC) and water-vapour adsorption. When the inlet-air temperature of the spray drier was increased beyond 140 degrees C, the intensity of X-ray diffraction peaks from crystalline UDCA decreased and the IR bands in the hydroxyl-stretching and carboxyl stretching regions changed. Dissolution of intact and spray-dried samples of UDCA prepared at 60 and 100 degrees C was an endothermic process but the dissolution became exothermic with increasing inlet-air temperature. UDCA samples differing in crystallinity were obtained, depending on the inlet-air temperature. A good correlation was obtained between the heat of solution and the heat of crystallization determined from DSC peak area. A good correlation was also obtained between the heat of solution and the crystallinity determined by Ruland's method from X-ray diffraction patterns. The amount of water vapour adsorbed on UDCA samples increased with increasing inlet-air temperature, indicating hydrogen bonding between water molecules and the hydroxyl groups or the carboxyl groups of amorphous UDCA. These results indicate that measurement of adsorption of water vapour and thermal analysis can both be used to evaluate the crystallinity of solid substances. PMID- 9877304 TI - Comparative measurement of the molecular weight of an antineoplastic glucan from BCG vaccine. AB - Bacillus Calmette-Guerin (BCG) vaccine, developed originally for the prophylaxis of tuberculosis, is a potent immunostimulant used to treat superficial bladder carcinoma in man. The aim of this study was to compare the molecular weight and self-association properties of an antineoplastic glucan (PS1A1) extracted from BCG vaccine as determined by different techniques including diffusion, light scattering and chromatographic methods. In the diffusion experiments, a semi empirical relationship was derived between the effective diffusion coefficients, Dp, and the weight-average molecular weights, Mw, of several dextrans used as standards, according to the equation Dp = 2.233 x 10(-6) x Mw(-0.66). On the basis of this relationship, the molecular weight of PS1A1 was found to be 57.4 kDa, although, unexpectedly, membrane association was high, most probably because of molecular branching. In the light-scattering experiment it was observed that, unlike dextran, PS1A1 undergoes concentration-dependent multimerization in water. However, the molecular weight of PS1A1 in 0.1 M sodium chloride ranged from 60 to 68 kDa, with a mean of 65 kDa, over the same concentration range. This value was in agreement with the molecular weight determined for PS1A1 by gel-filtration chromatography in previous studies, suggesting that 65 kDa represents the approximate monomeric size of the unassociated molecule. Thus, it was evident that the aggregation was suppressed by electrolyte. Elemental analysis by X-ray fluorescence showed that PS1A1 contained carbon, oxygen, hydrogen and phosphorus, indicating that hitherto unobserved ionized phosphate groups might promote electrostatic interactions. PMID- 9877302 TI - Alterations in rat vaginal histology by exogenous gonadotrophins. AB - Changes in vaginal epithelium are known to occur during the normal oestrous or menstrual cycle and in ovariectomised animals in response to various hormones. However, vaginal changes due to exogenous gonadotrophins superimposed on the normal hormonal milieu as occurs in in vitro fertilisation programmes have not previously been demonstrated. Female rats were hyperstimulated with follicle stimulating hormone and human chorionic gonadotrophin prior to mating. Control mated animals were not injected. Vaginal tissue was collected at 4.5, 5.5 and 6.5 d after mating. Tissue was processed for light microscopy. Hyperstimulation by the exogenous gonadotrophins caused mucification, a decrease in the number of epithelial layers and an increase in the thickness of the epithelium on d 4.5 of pregnancy. The mitotic index of the epithelial cells was depressed in the hyperstimulated rats with respect to control animals. Exposure to high levels of oestradiol and an altered progesterone:oestradiol ratio appear to have caused the changes in the vaginal epithelium. PMID- 9877306 TI - Submicron oil-in-water emulsion formulations for mefloquine and halofantrine: effect of electric-charge inducers on antimalarial activity in mice. AB - Stearylamine, oleic acid, phosphatidylserine and dicetylphosphate have been studied to determine their capacity to induce electric charge on non-ionic submicron emulsions containing halofantrine and mefloquine. The in-vivo antimalarial activity of drug-loaded emulsions, evaluated in mice, was affected by the nature of the additives used. In particular, the electric-charge inducers markedly affected the pharmacological activity of mefloquine, but not of halofantrine. After subcutaneous administration ED50 values (the doses affording 50% protection) were 3 and 15 mg kg(-1), respectively, for halofantrine and mefloquine emulsions without charge inducers. The mefloquine-loaded emulsions with charge inducers were active at 10 mg kg(-1) for dicetylphosphate, 17 mg kg( 1) for phosphatidylserine, 23 mg kg(-1) for oleic acid and 27 mg kg(-1) for stearylamine, again after subcutaneous administration. This work has enabled the formulation of stable emulsions, incorporating drugs with high antimalarial activity, which are proposed for parenteral delivery of these fairly soluble drugs. PMID- 9877307 TI - Enhanced absorption of calcium after oral administration of maltitol in the rat intestine. AB - The enhancing effects of maltitol (alpha-D-glucopyranosyl-1,4-sorbitol) on absorption of calcium by the rat intestine have been studied by use of [45Ca]CaCl2 in-vivo. After intragastric administration of [45Ca]CaCl2 solution with maltitol, plasma 45Ca concentration remained at the maximum level for more than 80 min, whereas for animals given [45Ca]CaCl2 solution without maltitol, plasma 45Ca concentration declined sharply after the peak. Determination of 45Ca radioactivity remaining in the various segments of the gastrointestinal tract revealed that administration of maltitol elicited slower gastric emptying and slower intestinal transit, resulting in extensive 45Ca distribution along the small intestine throughout the experimental period. The luminal contents of the small intestine were significantly higher in rats given maltitol than in the control group. These results suggest that the enhancing action of maltitol on intestinal calcium absorption could be attributed to reduced gastrointestinal calcium transit and increased luminal fluid content, presumably because of the osmotic activity of maltitol; this would not only accelerate the dissolution of calcium into the increased luminal contents, but also enable a larger area of the small intestine to absorb calcium for a longer period of time. PMID- 9877309 TI - Pharmacokinetic differences between lansoprazole enantiomers in rats. AB - Because limited information is available about potential differences between the pharmacokinetics and pharmacodynamics of the enantiomers of lansoprazole, the enantioselective pharmacokinetics of the compound have been investigated in rats. There was a noticeable difference between the serum levels of the enantiomers of lansoprazole and of their metabolites, 5-hydroxylansoprazole enantiomers, after oral administration of the racemate (50 mg kg(-1)) to rats. Cmax (maximum serum concentration) and AUC (area under the serum concentration-time curve) for (+) lansoprazole were 5-6 times greater than those for (-)-lansoprazole, whereas for (+)-5-hydroxylansoprazole both values were significantly smaller than those for the (-) enantiomer. CLtot/F values (where CLtot is total clearance and F is the fraction of the dose absorbed) for (+)-lansoprazole were significantly smaller than those for the (-) enantiomer. There was no significant difference between the absorption rate constants of the lansoprazole enantiomers in the in-situ absorption study. The in-vitro protein-binding study showed that binding of (+) lansoprazole to rat serum proteins was significantly greater than for the (-) enantiomer. The in-vitro metabolic study showed that the mean metabolic ratio (45.9%) for (-)-lansoprazole was significantly greater than that (19.8%) for the (+) enantiomer in rat liver microsomes at 5.6 microM lansoprazole. These results show that the enantioselective disposition of lansoprazole could be a consequence of the enantioselectivity of plasma-protein binding and the hepatic metabolism of the enantiomers. PMID- 9877308 TI - The effect of the mode of administration on the hypolipidaemic activity of niacin: continuous gastrointestinal administration of low-dose niacin improves lipid-lowering efficacy in experimentally-induced hyperlipidaemic rats. AB - The effect of different routes and modes of administration of niacin (nicotinic acid) on its hypolipidaemic activity has been evaluated. Our working hypothesis was that the major sites of niacin action are located presystemically (i.e. in the gut wall or the liver, or both) which would make niacin a gastrointestinal drug. For such drugs continuous administration to the gastrointestinal tract is expected to augment their efficacy compared with bolus oral administration or parenteral administration. The hypothesis was examined in two rat models of experimentally induced hyperlipidaemia-Model A, based on a cholesterol-enriched diet, and Model B, in which acute hyperlipidaemia is induced by intraperitoneal administration of triton (225 mg kg(-1)). Continuous administration of niacin into the duodenum at 1.66 mg h(-1) (total dose 40 mg kg(-1) day(-1)) for up to 7 days (Model A) or at 2.22 mg h(-1) over 18 h (Model B) had significantly greater lipid-reducing effects both on total cholesterol and on triglyceride levels (15 25%) and elevation of high-density lipoprotein (HDL) cholesterol levels than did bolus oral administration of the same dose. Continuous duodenal infusion of niacin also had an even greater lipid-reducing effect than continuous intravenous infusion of the drug at the same rate and dose. The results indicate that the site(s) of action are located presystemically and that continuous duodenal administration of a low dose of niacin (40 mg kg(-1)) has a greater lipid lowering effect than a higher dose (200 mg kg(-1)) administered by peroral bolus administration. These conclusions were validated by administration of a specially designed niacin sustained-release matrix tablet formulation that was non invasively administered to hyperlipidaemic rats. The hypolipidaemic activity of the sustained-release tablet was of similar magnitude to that resulting from continuous duodenal administration, thus providing a pharmacodynamic rationale for this mode of administration. PMID- 9877310 TI - Pharmacokinetics and hypotensive effect of diltiazem in rabbits: comparison of diltiazem with its major metabolites. AB - To assess the contribution of its metabolites to the antihypertensive effects of diltiazem, a previously established rabbit model has been used to compare the pharmacokinetics and haemodynamic effects of the drug with those of its major metabolites deacetyldiltiazem (M1) and deacetyl-N-monodemethyldiltiazem (M2). Diltiazem, M1 and M2 were administered separately to each animal (n = 5 or 6 per study group) as a single 5 mg kg(-1) intravenous dose. Blood samples, systolic and diastolic blood pressure (SBP and DBP) and heart rate were recorded for each rabbit up to 8 h, and urine samples were collected for 48 h post-dose. Plasma concentrations of diltiazem and its major metabolites were determined by HPLC. The results showed that systemic clearance (CL) and volume of distribution at steady state (Vdss) were smaller for diltiazem than for the metabolites. Diltiazem and the metabolites reduced both SBP and DBP, the effects of diltiazem being most potent. Their effects on heart rate were highly variable and not statistically different between treatment groups (P > 0.05). These results indicate that diltiazem is a more potent hypotensive agent than M1 or M2, possibly because of the higher plasma concentrations secondary to the smaller CL and Vdss of diltiazem compared with the metabolites. The effects of the metabolites might, however, be more sustained. PMID- 9877311 TI - The kinetic profile of vancomycin in neonates. AB - The pharmacokinetic parameters of vancomycin in a neonatal population have been characterized to enable development of optimum dosage guidelines for neonatal intensive-care units and to examine the relationship between these pharmacokinetic parameters and various demographic, developmental and clinical factors which might be associated with changes in the kinetic profile of vancomycin. Forty-four infants (twenty-five males and nineteen females) with suspected or proven Gram-positive infection and who received intravenous vancomycin between October 1993 and December 1996 were included in this retrospective analysis. Gestational age ranged from 25 to 40 weeks and postconceptional age at the time of the study ranged from 28 to 45 weeks. Sixty case-studies were obtained from the forty-four patients, with one period of study corresponding to one week or one cycle of therapy. Vancomycin pharmacokinetic parameters were determined by use of a one-compartment model. By regression analysis the current weight (g) was shown to be the stronger covariate, and both vancomycin clearance (L h(-1)) and volume of distribution (L) had to be normalized. The vancomycin volume of distribution depended on the postconceptional age with a cut-off at 32 weeks, whereas vancomycin clearance depended on the presence or absence of concomitant treatment with indomethacin or of mechanical ventilation, or both. On the basis of the pharmacokinetic parameters obtained we suggest initial dosage guidelines for vancomycin ranging from 10 mg kg(-1) every 8 h to 10 mg kg(-1) every 12 h, depending on the demographic and clinical characteristics of the patients. The results obtained enabled application of better a priori and a posteriori dosage schedules to infants in neonatal intensive-care units by use of the Bayesian approach, although further prospective study is recommended before direct extrapolation to patients in other settings. PMID- 9877312 TI - 6-Mercaptopurine pharmacokinetics after use of azathioprine in renal transplant recipients with intermediate or high thiopurine methyl transferase activity phenotype. AB - Prevention of allograft transplant rejection by the immunosuppressive 6 thiopurine drug azathioprine is limited by haematological toxicity (leucopenia or agranulocytosis). This toxicity is particularly apparent in subjects with low thiopurine methyltransferase activity (TPMTase) phenotype (1% in the Caucasian population). The thiopurine derivative 6-mercaptopurine is the active metabolite of azathioprine, and it would be of interest to measure, after validation of plasma measurements, the mean values of the pharmacokinetic parameters in transplant patients with high or intermediate TPMTase phenotypes (85 and 14% of the Caucasian population, respectively). We measured erythrocyte TPMTase activity in 103 kidney transplant recipients of high or intermediate phenotype and calculated, after a test dose of azathioprine, the mean values of the pharmacokinetic parameters for 6-mercaptopurine. We also compared these values with the same parameters from one subject with low TPMTase activity phenotype. The mean observed area under the plasma concentration-time curve (AUC) was 190+/ 140 ng mL(-1) h and the elimination rate constant (Kel) was 1.92+/-1. The pharmacokinetic parameters (AUC, Kel, t1/2el (the elimination half-life)) of 6 mercaptopurine in transplant patients are normally distributed and suitable for acceptance as a gold standard value for this population of Caucasian transplant patients. It seems useful to calculate these parameters, representative of the systemic exposure of individual patients to the drug, before prescribing these subjects azathioprine immunosuppressive treatment. In subjects with low TPMTase phenotype these pharmacokinetic measurements could also be an index of dose reduction. PMID- 9877313 TI - Alpha-adrenoceptor interaction of tetrandrine and isotetrandrine in the rat: functional and binding assays. AB - The action of 1S,1'S-tetrandrine, a bisbenzyltetrahydroisoquinoline alkaloid, on alpha1-adrenoceptors has been compared with that of its isomer 1R,1'S isotetrandrine. The work includes binding assays to analyse the affinity of these products for the [3H]prazosin binding site of rat cerebral cortical membranes and functional studies on rat isolated aorta to examine the effects of both alkaloids on intracellular calcium processes related or not to alpha-adrenoceptor activation. A radioligand receptor-binding study showed that both compounds interacted with the alpha1-adrenoceptors displacing [3H]prazosin from the specific binding site. The Ki values (inhibition constants) were 0.69+/-0.12 and 1.6+/-0.4 microM for tetrandrine and isotetrandrine, respectively. The functional studies showed that both alkaloids concentration-dependently inhibited noradrenaline-induced contraction in Ca2+-free solution (IC50 values, i.e. the concentrations needed to induce 50% inhibition, were 252.8 and 174.9 microM for tetrandrine and isotetrandrine, respectively), the spontaneous contractile response elicited by extracellular calcium after depletion of noradrenaline sensitive intracellular stores (increase in resting tone; IC50 values 11.6 and 19.6 microM for tetrandrine and isotetrandrine, respectively) and the refilling of intracellular Ca2+ stores sensitive to noradrenaline (IC50 values 7.4 and 14.9 microM for tetrandrine and isotetrandrine, respectively). The results show that tetrandrine and isotetrandrine interact with alpha1-adrenoceptors by displacing the [3H]prazosin binding site and that both compounds inhibit mainly the Ca2+ dependent process and have less action on alpha1-adrenoceptors. Tetrandrine is more potent than isotetrandrine. PMID- 9877314 TI - Effects of copper-aspirin complex on platelet aggregation and thrombosis in rabbits and mice. AB - The effects of intragastric and intraduodenal copper-aspirin complex on rabbit platelet aggregation were observed by Born's method. Myers's method was used to evaluate the antithrombotic effect of copper-aspirin complex in mice. In-vitro copper-aspirin complex selectively inhibited arachidonic acid-induced platelet aggregation with an IC50 value (concentration resulting in 50% inhibition) of 13.2 microM (95% confidence limits 9.1-16.8 microM). Copper-aspirin complex (10 mg kg(-1) given intragastrically or intraduodenally) was more potent than aspirin in inhibiting arachidonic acid-induced platelet aggregation. Copper-aspirin complex (10 mg kg(-1)) had a stronger inhibitory effect and a longer duration of action when given intragastrically than when given intraduodenally. It was shown by radioimmunoassay that copper-aspirin complex significantly reduced the level of thromboxane B2 in plasma while markedly increasing that of 6-ketoprostaglandin F1alpha (6keto-PGF1alpha). Copper-aspirin complex (10 mg kg(-1) given intragastrically for 7 days) significantly reduced mouse mortality caused by intravenous injection of arachidonic acid. The results suggest that both in-vitro and in-vivo copper-aspirin complex is more potent in selectively inhibiting arachidonic acid-induced platelet aggregation than aspirin. When given intragastrically the complex has a more potent antiplatelet effect and a longer duration of action than when given intraduodenally. The antithrombotic effect of the complex was more potent than that of aspirin. PMID- 9877315 TI - Protein kinase inhibitors attenuate cardiac swelling-induced amino acid release in the rat. AB - Rat Langendorff heart preparations have been used to study the efflux of cardiac amino acids into coronary artery perfusates during brief (5-min) periods of exposure to hyposmotic stress (70 mM NaCl). Coronary flow rates, heart rates and intra-aortic pressures were recorded. Amino acid levels were measured by high performance liquid chromatography. Hyposmotic stress caused marked percentage increases in taurine, glutamate and aspartate levels in the coronary perfusate, with smaller increases in phosphoethanolamine, glycine and alanine and non significant increases in serine and glutamine. Amino acid levels declined during reperfusion with isosmotic Krebs-Henseleit bicarbonate buffer. Inhibition of protein kinase C with chelerythrine chloride (5 microM) depressed the osmotically induced release of aspartate, glutamate, taurine and glycine. The protein tyrosine kinase inhibitor, genistein, reduced the anisosmotic efflux of aspartate, glutamate, taurine and phosphoethanolamine. Lavendustin A, another inhibitor of tyrosine kinase, depressed the osmotically evoked release of aspartate, glutamate and taurine. These studies demonstrate the involvement of protein kinase C and tyrosine kinases in the efflux of amino acids from the osmotically challenged rat heart and imply that these enzymes are involved in the mechanisms responsible for volume regulation by cardiac cells. PMID- 9877317 TI - Ouabain-induced increase in dopamine release from mouse striatal slices is antagonized by riluzole. AB - We have examined the effects of riluzole, a neuroprotective drug which stabilizes voltage-dependent sodium channels in their inactivated state and inhibits the release of glutamate in-vivo and in-vitro, on the release of newly taken up [3H]dopamine induced by ouabain, a potent and selective inhibitor of Na+/K+ ATPase in mouse striatal slices in-vitro. Riluzole potently (IC50 (concentration resulting in 50% inhibition) = 0.9+/-0.3 microM) and dose-dependently antagonized ouabain-stimulated [3H]dopamine release, the effect being observed at low concentrations. Tetrodotoxin (1 microM) and nomifensine (10 microM) also abolished ouabain-induced [3H]dopamine release. Blockade of glutamate receptors with dizocilpine (1 microM) and 6-(1H-imidazol-1-yl)-7-nitro-2,3(1H,4H) quinoxalinedione (YM-90K; 10 microM), alone or in combination, was without effect. Incubation of striatal slices with 50 microM La3+, which blocks voltage dependent calcium channels, did not inhibit [3H]dopamine release induced by ouabain. The potent effects of riluzole observed in this model are probably related to its ability to block voltage-dependent sodium channels. The consequences of this activity are critically discussed in relation to the protective action of riluzole previously reported in various models of Parkinson's disease and other neurodegenerative disorders. PMID- 9877316 TI - Pharmacological antagonism of Anchietia salutaris extracts on the contraction induced by prostaglandin D2 and U46619 in guinea-pig lung parenchymal strips. AB - Anchietia salutaris tea is traditionally used in Brazil to treat allergies, suggesting it contains compounds with antagonistic activity on the allergic mediators. We have evaluated extracts and semi-purified fractions of Anchietia salutaris as a source of compounds having this type of antagonism on the contraction induced in guinea-pig lung parenchymal strips and on platelet aggregation and shape change. After 10 min pre-incubation dichloromethane extracts containing 30 or 100 microg mL(-1) inhibited the contraction induced by prostaglandin D2 (PGD2) in guinea-pig lung parenchymal strips with dose ratios (DR) of 0.76+/-0.14 and 0.93+/-0.19, respectively; the amount of inhibition depended both on the concentration and on the time of pre-incubation (DR after 30 min pre-incubation was 1.21+/-0.51). The dichloromethane extract and its semi purified fractions also inhibited the contractions induced by U46619, a more potent, stable, synthetic agonist of thromboxane A2 (TxA2) prostanoid (TP) receptors, the receptors acted upon by PGD2 to produce lung contractions. The dichloromethane extract did not inhibit the lung parenchymal contractions induced by histamine, leukotriene D4 (LTD4) or platelet-activating factor (PAF). Platelet aggregation induced by U46619, adenosine 5'-diphosphate (ADP) or PAF was not inhibited by the dichloromethane extract. Indeed, the extract potentiated platelet aggregation induced by low concentrations of these agonists and also potentiated the shape change induced by U46619. These results imply that the dichloromethane extract of Anchietia salutaris and its semi-purified fractions contain an active principle that competitively inhibits TxA2 TP receptors, the stimulation of which causes lung parenchymal contraction. The inhibition seems to be selective for this receptor subtype, because the extract fails to inhibit platelet aggregation or shape change. This provides additional support of earlier reports suggesting the occurrence of TP receptor subtypes. PMID- 9877319 TI - The role of the iminium bond in the inhibition of reverse transcriptase by quaternary benzophenanthridines. AB - The quaternary benzo[c]phenanthridines fagaronine, nitidine and O methylfagaronine have been reviewed as potential antitumour and antiviral agents. Their mode of action has not been established, but their ability to bind with DNA by intercalation is believed to be involved. Of the three synthetic analogues of O-methylfagaronine which we have synthesized, methoxidine and ethoxidine are active against HIV-1 reverse transcriptase (IC50 values 2.8 microM and 2.4 microM respectively) whereas hydroxidine is inactive. One of the prerequisites for the enzyme inhibitory activity of this class of molecule is the presence of an iminium group--it is well known that a positive charge on a polyaromatic nucleus facilitates intercalative binding with DNA. Through UV spectrophotometric and modelling studies, we have shown that the iminium bond plays a more fundamental role in enzyme inhibition through its susceptibility to nucleophilic attack--the inactive analogue hydroxidine has a non-electrophilic iminium bond. Consequently, we have demonstrated that iminium bond electrophilicity is a parameter which needs to be considered in ternary complex formation with reverse transcriptase. PMID- 9877318 TI - Anti-nicotinic properties of anticholinergic antiparkinson drugs. AB - The nature of the antagonism by anticholinergic compounds of nicotine-induced convulsion in mice has not been defined clearly. Although, because they do not compete effectively for agonist binding to brain tissue in-vitro, these compounds are thought to be non-competitive antagonists in the brain, pharmacological evidence is lacking. This study describes the anti-nicotinic properties of the clinically used anticholinergic antiparkinson drugs, benztropine, biperiden, caramiphen, ethopropazine, procyclidine and trihexyphenidyl. Nicotine-induced convulsion and arecoline-induced tremor in mice were effectively prevented by these drugs. The concentrations of benztropine, biperiden, caramiphen, ethopropazine, procyclidine and trihexyphenidyl affording 50% prevention of nicotine-induced convulsion (ED50 values) were 7.4, 4.6, 7.8, 4.9, 3.1 and 3.3 mg kg(-1), respectively. The classical muscarinic receptor antagonist atropine had potent anti-muscarinic effects but very weak anti-nicotinic activity. The classical nicotinic receptor antagonist mecamylamine had potent anti-nicotinic activity but no anti-muscarinic effects. The pattern of shift of the dose response curve for nicotine-induced convulsion in mice was determined in the presence of increasing concentrations of the anticholinergic antiparkinson drugs. These drugs were found to increase the ED50 (0.49 mg kg(-1)) of nicotine-induced convulsion in a dose-related manner. The maximum effect of nicotine and the slope of nicotine dose-response curve were not significantly influenced by either low or high doses of benztropine, procyclidine or trihexylphenidyl, which suggests competitive action. Biperiden, caramiphen and ethopropazine, at low doses which significantly increased the ED50 of nicotine, did not affect the maximum effect of nicotine or the slope of the nicotine dose-response curve; at higher doses, however, they reduced the maximum effect and the slope, which suggests that these drugs have both competitive and non-competitive properties in antagonizing nicotine-induced convulsion in mice. The experiments demonstrate that the anticholinergic antiparkinson drugs and mecamylamine effectively antagonize nicotine-induced convulsion, but atropine does not; some of these drugs have competitive properties whereas others seem to have both competitive and non competitive properties in antagonizing nicotine-induced convulsion in mice. PMID- 9877320 TI - Rapid killing of Mycobacterium terrae by N-chlorotaurine in the presence of ammonium is caused by the reaction product monochloramine. AB - We have studied the activity of the weak endogenous oxidant N-chlorotaurine against Mycobacterium terrae. The study revealed slow killing of more than 2h duration by 1% (55 mM) N-chlorotaurine. In the presence of ammonium chloride, however, killing times decreased to a few minutes, even by 0.1% N-chlorotaurine. This phenomenon is explained by formation of the lipophilic and therefore more bactericidal monochloramine as a result of transhalogenation of ammonia by N chlorotaurine. PMID- 9877321 TI - Enantioselectivity of some 1-(benzofuran-2-yl)-1-(1-H-imidazol-1-yl) alkanes as inhibitors of P450Arom. AB - The low stereospecificity of the enantiomers of 1-[(benzofuran-2-yl)-4 chlorophenylmethyl]imidazole (6, R=H, R'=4'-Cl) and the corresponding 4-fluoro compound as inhibitors of aromatase (P450Arom) has been explored using 1-(5,7 dichlorobenzofuran-2-yl)-1-(1H-imidaz-1-yl)ethane (7, R1=R2=Cl, R=CH3), -propane (7, R1=R2=Cl, R=C2H5), and the corresponding 5,7-dibromo compounds resolved as their dibenzoyl-D (or -L) tartrates. Low enantioselectivity ratios of 4.8 (5,7 diCl) and 12.6 (5,7-diBr) were shown for the ethanes. The values for the corresponding propanes were 8.3 and 5.2, respectively, and for these compounds the stereoselectivity was reversed. PMID- 9877323 TI - Transcranial magnetic-stimulation mapping of the cortical topography of the human masseter muscle. AB - The cortical topography of several limb and trunk muscles has been disclosed using transcranial magnetic stimulation, but the corticomotor representation of the human jaw muscles has not yet been described. An experimental paradigm incorporating transcranial magnetic stimulation of verified sites on the scalp was used (a) to determine the cortical topography of the human masseter muscle and (b) to assess the reproducibility of the motor map, in seven healthy individuals. The results showed that the masseter was discretely represently on the motor and premotor cortex with interindividual variation in map area, volume and height (p < 0.05). Coefficients of variation and reliability were low (15 18%) and high (89-96%), respectively, for motor maps obtained at experimental sessions 2 weeks apart, indicating high reproducibility. The findings have clinical relevance in the management of patients with cerebral injury involving corticobulbar projections to the masseter. PMID- 9877322 TI - The effects of fluoride concentration and the level of cariogenic challenge on caries development in desalivated rats. AB - Dental caries is an infectious and transmissible disease that continues to affect the majority of people. The presence of carbohydrate, mainly sucrose in the diet, is an important factor in its occurrence. The amount of fluoride required for optimal protective effect where there is a high caries challenge is unclear. Differences in the intensity of cariogenic challenge, for whatever reason, may play a part in determining fluctuations in the effectiveness of fluoride. The purpose of this study was to evaluate the effect of different concentrations of fluoride on the development of caries and explore the cariostatic effect of fluoride under various levels of cariogenic challenge. The study comprises two experiments. In experiment I, 60 desalivated Sprague Dawley rats infected with Streptococcus sobrinus were offered the following to drink for 21 days: group (1), sterile distilled water (SDW); (2) 10 parts/10(6) F SDW; (3) 20 parts/10(6) F SDW; (4) 30 parts/10(6) F SDW; (5) 40 parts/10(6) F SDW. In experiment II, eight groups of 9 rats were placed in a Konig Hofer programmed feeder and were exposed to different levels of cariogenic challenge through varying frequency of eating and offered water containing 10 parts/10(6) F. In experiment I, exposure to 20, 30 and 40 parts/10(6) F reduced caries development significantly: fluoride, at 10 parts/10(6), reduced the severity of the carious lesions. In this model of severe cariogenic challenge, the results suggest that elevated concentrations of fluoride might be effective in patients at high caries risk. In experiment II, fluoride reduced the incidence and severity of smooth-surface caries in all groups. The protective effect of fluoride decreased as the number of exposures to sugar increased. It is concluded that the effectiveness of fluoride is influenced by the level of cariogenic challenge and that consideration should be given to adjusting the level of fluoride exposure based on perceived caries risk, and that there is a maximum therapeutic effect of fluoride beyond which no additional protection can be expected. PMID- 9877325 TI - Calculus deposits and bone loss on the teeth of Romano-British and eighteenth century Londoners. AB - The relation between dental calculus and periodontal disease is not clear but it is generally recognized that calculus is a significant pathogenetic factor. Skeletal material has previously been used to study some aspects of chronic adult periodontitis but few studies have quantified the extent of calculus in ancient populations and its relation to changes in alveolar bone height. This study records the presence and extent of calculus and its relation to alveolar bone loss in a Romano-British and eighteenth-century London population. There were significant differences in calculus deposition in the two populations but this appeared to have little effect on changes in alveolar bone contour. It is suggested that the amount of calculus may be related to diet but that changes in alveolar bone height seem to be independently controlled. PMID- 9877324 TI - Modification of the beta- and alpha2-adrenergic sensitivity of rat submandibular glands by environmental stimuli and stress. AB - In man, the rate of resting salivary secretion can be influenced by environmental stimuli related to light dark cycles or by noxious stimuli (stressors) of psychological origin. The sympathetic branch of the autonomic nervous system and the adrenal medulla play an important part in homeostatic responses. Previous observations have shown that chronic exposure of rats to constant light promotes degranulation of parotid acini and desensitization of submandibular beta adrenergic receptors. Now the submandibular secretory response elicited by beta- and alpha2-adrenergic agonists was studied in rats chronically exposed to environmental conditions that modified the activities of sympathetic efferents to the pineal, salivary and adrenal glands. Adult male rats were exposed to constant light (LL) or constant darkness (DD) for 20 days, or to stress (2 h daily immobilization) for 14 days. Control animals were kept under the usual lighting conditions and without immobilization. Dose response curves to isoproterenol (i.v), before and after administration (i.v.) of a dose (20 microg/kg) of clonidine were obtained. Beta-adrenergic desensitization was observed in all the experimental groups, while alpha2-adrenergic desensitization was only observed in the stress and LL groups. The results suggest that circulating catecholamines could mediate light and stress effects on submandibular beta-adrenergic secretory responses. Extrasynaptic alpha2-adrenoceptors might modulate the submandibular secretory response when predictable environmental stimuli (daily light phase) or unpredictable stressors raise the concentrations of circulating catecholamines. PMID- 9877327 TI - Complete amino acid sequences and phosphorylation sites, determined by Edman degradation and mass spectrometry, of rat parotid destrin- and cofilin-like proteins. AB - Beta-adrenergic or cholinergic stimulation of the rat parotid gland was earlier shown to induce dephosphorylation of endogenous destrin- and cofilin-like proteins, which are phosphorylated in resting cells at Ser residues probably present near the N-terminals. The primary structures and phosphorylation sites were determined here. The rat destrin-like protein had a sequence 95% identical to the cDNA-derived sequence of porcine destrin. The rat cofilin-like protein was 98% identical to that of porcine cofilin. Each protein lacked the initiator Met and began with an acetylalanine residue followed by a Ser residue. The N-terminal peptides generated with endoproteinase Asp-N were isolated; they were each phosphorylated at Ser-2. Earlier work had shown that partial cleavage of the phosphorylated destrin- and cofilin-like proteins with cyanogen bromide provides unphosphorylated 16.7- and 18.3-kDa fragments, respectively. It was here confirmed that they contained all the Ser residues other than those present in the N-terminal peptides. From these observations, it was now concluded that the destrin- and cofilin-like proteins are rat parotid destrin and cofilin (non muscle type), respectively, and that each protein is phosphorylated exclusively at Ser-2 in resting cells and dephosphorylated at this site in response to beta adrenergic or cholinergic stimulation. PMID- 9877326 TI - Modulation by presynaptic beta-adrenoceptors of noradrenaline release from sympathetic nerves in human dental pulp. AB - This study was undertaken to test for the presence of presynaptic beta adrenoceptors on sympathetic nerves in human dental pulp and, if present, to investigate the subtype. Pulp was excised from freshly extracted teeth, incubated with [3H]-noradrenaline (0.6 micromol/l) and subsequently superfused with Krebs solution. Sympathetic nerves were stimulated at 5 Hz for 100 sec. The non specific beta-adrenoceptor agonist isoprenaline (1.0 micromol/l), and the selective beta2-agonist salbutamol (10 micromol/l) facilitated the release of [3H]-noradrenaline; isoprenaline, but not salbutamol, also facilitated this release in the presence of desipramine (DMI, 0.3 micromol/l), corticosterone (10 micromol/l) and rauwolscine (0.1 micromol/l). BRL 37344 (1.0 micromol/l), a beta3 agonist, had no effect on [3H]-noradrenaline release. The facilitatory effects of isoprenaline and salbutamol were inhibited by the non-specific beta-antagonist propranolol (1.0 micromol/l), while that of salbutamol was inhibited in the presence of ICI-188,551 (1.0 micromol/l), a selective beta2-antagonist, as well. The beta1-antagonist atenolol (1.0 micromol/l) potentiated the facilitatory effects of isoprenaline in the presence of DMI and corticosterone. Neither propranolol nor ICI-188,551 alone affected the release of [3H]-noradrenaline. These results establish the presence of presynaptic beta-adrenoceptors on sympathetic nerves in human dental pulp. It is suggested that they are of the beta2-subtype, although a greater range of agonists and antagonists needs to be used to clarify the nature of the the beta-adrenoceptors. PMID- 9877328 TI - Histogenesis of the chequered pattern of ivory of the African elephant (Loxodonta africana). AB - This study aimed to propose a hypothesis on the events which lead to the development of the characteristic chequered pattern of elephant ivory. Twenty fragments of ivory and six elephant tusks were obtained through the National Parks Board of South Africa. Polished surfaces were prepared in sagittal and longitudinal planes and the characteristics of the distinctive chequered pattern described. Light- and electron-microscopical techniques and image analyses were employed to determine the morphological basis of the pattern and to describe the spatial distribution, density and morphology of the dentinal tubules. These investigations showed that the distinctive pattern was the result of the sinusoidal, centripetal course followed by dentinal tubules. The apical, slanted part of the sinusoidal curve is the result of the centripetally moving odontoblast, which, during formation of ivory, progresses towards the centre of the tusk on a decreasing circumference. It is suggested that this leads to cell crowding, increased pressure between odontoblasts and subsequent apical movement of their cell bodies, cell degeneration and fusion. Odontoblastic degeneration and fusion probably relieve the pressure between the crowded odontoblasts by reducing their numbers and the remaining odontoblasts now orientate their centripetal course towards the tip of the tusk, thereby forming the anterior directed part of the sinusoidal path of the tubule. As odontoblasts progress centripetally the diameter of the pulpal cavity decreases further and the processes of apical movement, fusion and degeneration of odontoblasts are repeated. This occurs until the pulpal cavity is obliterated. PMID- 9877329 TI - Inhibition of the synthesis of eicosanoid-like substances in a human oral cancer cell line by interferon-gamma and eicosapentaenoic acid. AB - The objectives were to examine the production of eicosanoids in a Chinese human oral cancer cell line (OEC-M1) and to test the effects of interferon-gamma (IFN gamma), eicosapentaenoic acid (EPA) and enzyme inhibitors on this biosynthesis. The eicosanoids were identified by reverse phase-high performance liquid chromatography. Two predominant peaks appeared in the chromatograms. One compound (P-1) was identified by ultraviolet absorption at a lambda(max) of 278nm with shoulders at 272 and 284nm. The other compound (P-2) was identified by ultraviolet absorption at a lambda(max) of 284 nm with shoulders at 278 and 290 nm. The production of P- was significantly inhibited by the addition of IFN-gamma (200 and 400 U/ml), and EPA (10 to 40 microM). It was only partially inhibited (p < 0.05) by indomethacin (INDO) (0.5 and 1 microM), nordihydroguaiaretic acid (NDGA) (30 and 60 microM/ml), and eicosa-5,8,11,14-tetraynoic acid (ETYA) (20-60 microM). It was almost completely inhibited by indomethacin (2 and 3 microM), and dexamethasone (0.6 and 6 microM). The production of P-2 was almost completely inhibited by IFN-gamma (200 and 400 U/ml), and partially inhibited (p < 0.05) by EPA (10 and 20 microM), NDGA (30 and 60 microM), ETYA (20 and 40 microM), dexamethasone (0.6 and 6 microM). The production of both peaks was significantly reduced by excluding arachidonic acid (AA), and almost completely inhibited by heating at 100 degrees C for 10 min during incubation. These results demonstrate that two eicosanoid-like compounds are synthesized by the OEC-M cell line and that their production can be modulated by IFN-gamma, EPA, indomethacin, NDGA, ETYA, and dexamethasone. PMID- 9877331 TI - Adhesion of oral Candida albicans isolates to denture acrylic following limited exposure to antifungal agents. AB - Candidal adherence to denture acrylic surfaces is implicated as the first step in the pathogenesis of Candida-associated denture stomatitis, the most prevalent form of oral candidosis in the West. This condition is treated by topically administered antifungal agents, mainly belonging to the polyenes and azoles. As the intraoral concentrations of antifungals fluctuate considerably due to the dynamics of the oral environment, the effect of short exposure to sublethal concentrations of antifungals on the adhesion of Candida albicans to denture acrylic surfaces was investigated. Seven oral C. albicans isolates were exposed to four-eight times minimum inhibitory concentrations (MIC) of five antifungal drugs, nystatin, amphotericin B, 5-fluorocytosine, ketoconazole and fluconazole, for 1 h. After removing the drug (by repeated washing) the adhesion of these isolates to acrylic strips was assessed by an in vitro adhesion assay. Exposure to antifungal agents significantly reduced the adherence of all seven C. albicans isolates to denture acrylic. The mean percentage reductions of adhesion after limited exposure to nystatin, amphotericin B, 5-fluorocytosine, ketoconazole and fluconazole were 86.48, 90.85, 66.72, 65.88 and 47.42%, respectively. These findings indicate that subtherapeutic doses of antifungals may modulate oral candidal colonization. Further, these results may have an important bearing on dosage regimens currently employed in treating oral candidosis. PMID- 9877330 TI - Role of epidermal growth factor and its receptor in mechanical stress-induced differentiation of human periodontal ligament cells in vitro. AB - The periodontal ligament (PDL) contains precursor cells for osteoblasts and cementoblasts. It has been shown that epidermal growth factor (EGF) inhibits dexamethasone-induced differentiation and up-regulates EGF-receptor (EGF-R) expression, whereas EGF-R is down-regulated in the course of differentiation. Thus it was suggested that EGF and its receptors act as a negative regulator of osteoblastic differentiation in PDL cells. In order to investigate further this hypothesis, human PDL cells were now used to elucidate the role of EGF and EGF-R in their proliferation and differentiation under mechanical stress-loaded conditions in vitro, as the PDL regularly receives mechanical stress from occlusal forces. As a model of mechanical stress, a cyclic stretch of 9 or 18% elongation was applied to the cells with a Flexercell cell-strain unit system. Alkaline phosphatase activity and osteocalcin mRNA expression were significantly induced by loading cyclic stretch for more than 4 days, whereas stretch slightly inhibited cell proliferation. Visualization of the actin stress fibres of the cells by rhodamine phalloidin revealed that approx. 10% of the total number of cells had become aligned perpendicularly to the direction of the stretch. The effects of stretch on alkaline phosphatase activity and cell proliferation were totally abolished by the presence of 10 ng/ml EGF. Western blotting of EGF-R protein demonstrated that stretch-induced differentiation accompanied the decreased expression of EGF-R protein in the cells. However, the amount of tyrosine-phosphorylated EGF-R upon EGF stimulation was restored to the control level in stretched cells. These results suggest that the EGF/EGF-R system acts as a negative regulator of differentiation of PDL cells regardless of the type of differentiation stimuli. Also, interaction between mechanical stress and the EGF/EGF-R system may participate in the osteoblastic differentiation of PDL cells and thereby regulate the source of cementoblasts and osteoblasts. PMID- 9877332 TI - Comparative observations on the spacing of short-period (von Ebner's) lines in dentine. AB - The spacing of short-period incremental markings in dentine was measured in longitudinal ground sections and in longitudinal demineralized silver-stained sections of permanent human canines and premolars. Measurements were made (i) within 50 microm from the granular layer of Tomes (GLT), (ii) between 100 and 200 microm from the GLT, and (iii) in the axial plane of the tallest cusps. Median values for the spacing of calcospheritic lines closest to the GLT in the ground sections increased from 1.8 to 2.8 microm as the lines gradually coalesced into a laminar pattern beyond the GLT pulpally. Median values for the spacing of short period lines in the cuspal dentine, where dentine formation is known to be fastest, were 4.1 microm. Markings in the demineralized sections were between 25 and 39% closer together, presumably due to contraction and shrinkage during specimen preparation. The spacings of short-period incremental lines measured on ground sections of non-human primate dentine (gibbon, siamang, orang) and on pig dentine, all between 100 and 200 microm from the GLT, clustered between 2.5 and 3.5 microm. Apart from gibbon dentine (in which spacings were closer together in this position than in the others), the distribution of measurements was not significantly different in pig, orang or human dentine. However, none of the data for the comparative samples presented here revealed spacings of short-period lines anywhere close to the 16 microm previously reported for circumpulpal dentine in animals. These data suggest that there may be many other animals where the mode and, to some extent, rate of dentine mineralization close to the root surface follows a common pattern. While data for the spacing of incremental markings in dentine provide no evidence for their periodicity, it is clear that the measurements made in the ground sections match the reported daily rates of mineralization at these locations, whereas those in demineralized silver-stained sections do not. Tissue shrinkage is probably a better explanation for this than the generally accepted view that they represent 12 h increments of dentine mineralization. This study provides a better basis for identifying and describing these lines, and for distinguishing them from other kinds of incremental markings in dentine. PMID- 9877333 TI - Variation of salivary calcium, phosphate and buffering capacity in adolescents. AB - Calcium, phosphate and buffering capacity were measured in children's (n=43) unstimulated saliva between September 1990 and June 1991 inclusive. Saliva samples were collected in the afternoon, once a month, on 10 consecutive occasions. Buffering capacity was assessed immediately after sample collection using the Dentobuff strip method. The remainder of the saliva sample was then stored at -18 degrees C and, after thawing, total calcium and inorganic phosphate were measured. The intra-individual variation of buffering capacity over time was statistically significant (Friedman two-way ANOVA, p < 0.001). The intra individual variability of salivary calcium was also statistically significant (repeated-measures ANOVA, p < 0.001). However, no evidence of a statistically significant variation in phosphate was observed. It is concluded that a combination of flow-rate fluctuation over time and the methods used to assess salivary constituents might have contributed to this variability. PMID- 9877335 TI - Revealed identity: a study of the process of genetic counselling. AB - This paper describes a qualitative study of the processes employed within a genetic counselling consultation. These processes, it is argued, can be seen to provide the patient with a genetic identity. But unlike the new identity conferred by the diagnosis of many chronic medical conditions a genetic identity is presented as an old one that is now revealed. This represents a reversal of the stigmatising process: instead of a spoiled identity replacing an actual one, the genetic consultation involves revealing an actual identity in the place of a previously presumed one. In this way, genetic counselling reconstructs identity in the past as well as in the future. PMID- 9877334 TI - Screening and counselling for sickle cell disorders and thalassaemia: the experience of parents and health professionals. AB - Shortfalls in haemoglobinopathy provision result in patients and their carers receiving inadequate support. This paper, by drawing on material from a project evaluating service provision to families caring for a child with a sickle cell disorder or thalassaemia, discusses screening and counselling services. It explores the perspectives of parents, front-line practitioners, managers and health commissioners. Poor quality care, inadequate information and professionals' insensitivity were salient themes in parental accounts. The parents' experience also confirms the problems faced by minority ethnic people in having their welfare needs recognised, more generally. Although our focus in on genetic conditions affecting minority communities in the UK, the issues we address are at the heart of the 'new genetics'. PMID- 9877337 TI - Educational differences in leisure-time physical inactivity: a descriptive and explanatory study. AB - In this study we aim to explain educational differences in leisure-time physical inactivity in terms of psychosocial and material factors. Cross-sectional data were obtained from the baseline of the Dutch GLOBE study in 1991, including 2598 men and women, aged 15-74 years. Physical inactivity during leisure time was defined as not participating in any activity, such as sports, gardening, walking or cycling. Psychosocial factors included in the study were coping resources, personality, and stressors. Material factors were financial situation, employment status, and living conditions. Logistic regression models were used to calculate educational differences in physical inactivity. Physical inactivity was more prevalent in lower educational groups. Psychosocial factors related to physical inactivity were locus of control, parochialism, neuroticism, emotional social support, active problem focussing, optimistic and palliative coping styles. Material factors associated with physical inactivity were income, employment status and financial problems. All correlates of physical inactivity were unequally distributed over educational groups, except optimistic and palliative coping. Personality and coping style were the main contributors to the observed educational differences in physical inactivity. That is to say, parochialism, locus of control, neuroticism and active problem focussing explained about half of elevated odds ratios of physical inactivity in the lower educational groups. The material factors, equivalent income and employment status explained about 40% of the elevated odds ratios. Psychosocial and material correlates together reduced the odds ratios of lower educational groups by on average 75%. These results have practical consequences for the design of more effective interventions to promote physical activity. In particular, personality and coping style of risk groups, such as lower educational groups, should be taken into consideration at the future development of these interventions, as well as inequalities in material restrictions related to engaging in physical activity. Supplementary interventions focussing on childhood conditions which, partly, influence both personality and physical inactivity may also contribute to a reduction of socio-economic differences in physical inactivity. PMID- 9877338 TI - Suicide in urban New South Wales, Australia 1985-1994: socio-economic and migrant interactions. AB - Variation of suicide with socio-economic status (SES) in urban NSW (Australia) during 1985-1994, by sex and country or region of birth, was examined using Poisson regression analysis of vital statistics and population data (age >? approximately 15 yr). Quintiles of SES were defined by municipality of residence and comparisons of suicide by SES were adjusted for age and country (or region) of birth (COB), and examined by COB. Risk of suicide in females was 28% that of males for all adults and 21% for youth (age 15-24 yr). Suicide risk was lower in males from southern Europe, Middle East and Asia, and higher in northern and eastern European males, compared to the Australian-born. Risks for suicide increased significantly with decreasing SES in males, but not in females. The relationship of male suicide and SES was stronger when controlled for COB. For males, the relative risk of suicide, adjusted for age and COB, was 66% higher in the lowest SES quintile compared to the highest quintile. and 39% higher for youth (age 15-24 yr). For male suicide, the population attributable fraction for SES (less than the highest quintile) was 27%. Analysis of SES differentials in male suicide according to COB indicated a significant inverse suicide gradient in relation to SES for the Australian-born and those born in New Zealand and the United Kingdom or Eire, but not in non-English speaking COB groups, except for Asia. For Australian-born males, suicide risk was 71% higher in the lowest SES group (compared to the highest), adjusted for age. These findings indicate that SES plays an important role in male suicide rates among the Australian-born and migrants from English-speaking countries and Asia, and among youth; but not in female suicide, nor suicide in most non-English speaking migrant groups. Reduction in SES differentials through economic and social policies may reduce male suicide in lower SES groups and should be seen to be at least as important as individual level interventions. PMID- 9877339 TI - Hearts and minds: suicide among United States combat troops in Vietnam, 1957 1973. AB - The literature on suicides among military personnel in a combat zone remains anecdotal. Although one finds literary and journalistic accounts of the problem, it has not been systematically analyzed. This paper will examine suicides among American combat troops during their tour of duty in Vietnam. Utilizing statistical data from the Southeast Asia Combat Area Casualties Database (SACACD), the authors will survey its epidemiology among U.S. ground troops in Vietnam from 1957 to 1973. The results suggest that certain types of combat troops were significantly more likely than others to commit suicide. These findings not only provide important predictors for military health professionals in the assessment of suicide risk, they also raise disturbing questions about the nature of organized armed conflict. PMID- 9877340 TI - Increasing homicide rate in Finland accompanied by decreasing seasonality over the period 1957-95. AB - Homicide is recognized as a global public health problem. Finland has the highest overall homicide rate of the Nordic countries in comparisons over short time periods. Using the 39-year time series of homicide incidents in Finland from 1957 95, we analyzed the trend in homicide rate. In addition, we explored for the first time over the time trend in the seasonal variation of homicide and compared seasonal statistics over eight successive time periods. The present study revealed that there has been steadily increasing trend in homicides in Finland since 1950s. At the same time, the seasonality of homicide has decreased markedly. The peaks in homicide rates occurred commonly during summer and the troughs during winter. PMID- 9877336 TI - Effect of social class on tumour size at diagnosis and surgical treatment in Danish women with breast cancer. AB - This study examines the effect of socio-economic inequalities on the tumour size at diagnosis and the choice of surgical treatment in Danish women with breast cancer. The Danish Breast Cancer Cooperative Group (DBCG) registers all women with breast cancer in Denmark and provides clinical register data on tumour size and surgical treatment for 1594 women <75 yr diagnosed with breast cancer between 1991 and 1996. A questionnaire including questions on social class was sent to the women. The tumour size at diagnosis (an indicator of access to treatment) and the surgical procedure (lumpectomy or mastectomy) for patients who were potential candidates for lumpectomy were the main outcome measures examined. Social class was not associated with tumour size at diagnosis. The tumour size was associated with age and the existence of a mammographic screening program in the county. However, treatment was strongly related to social class. Among women considered candidates for lumpectomy 77% underwent lumpectomy in the highest social class, compared to 50% in the lowest social class. Treatment was associated with age, tumour size and hospital status. It was concluded that the study showed no socio economic inequalities related to access to health services as measured by tumour size, though significant social differences were found regarding treatment. PMID- 9877342 TI - Using metaphor to read the organisation of the NHS. National Health Service. AB - The paper is concerned with the complexity of the British National Health Service (NHS) as an organisation and with different ways of seeing this. Morgan proposes that explanations of organisational life are based on metaphors which highlight particular interpretations (Morgan, G., 1986, Images of Organisation. London, Sage). The ability to "read" a complex phenomenon depends on being able to see how these different aspects co-exist. The study applies metaphoric thinking to the organisation of the NHS. Utilising documentary data sources, a diagnostic reading is made examining different metaphors to highlight key aspects of the situation. The metaphors of machine and organism are drawn from Morgan's conceptual scheme, those of religion and marketplace are proposed as of particular relevance to the NHS. In the religious metaphor the focus is on the mission of the NHS in terms of its founding principles of universality, comprehensiveness, equality and collectivism. Perceived as a machine the NHS is characterised as an organisation originally based on technocratic rationality and its subsequent history interpreted as moving towards increasingly centralised control. An alternative perspective on the same events is considered in terms of the organic metaphor. In this view the NHS is examined as an open system, which is devolved, decentralised, participative and responsive to its environment. The image of the marketplace focuses on the impact on the organisation of the introduction of competition and incentives in the post-reform period. Other images are sketched briefly. In the critical evaluation the insights generated by the different images are assessed and the different interpretations linked together. It is concluded that metaphoric thinking enables us to appreciate and interpret the ambiguities and paradoxes in NHS organisational life. PMID- 9877343 TI - Achieving clinical behaviour change: a case of becoming indeterminate. AB - This paper is based on an empirical study of attempts to achieve change in clinical behaviour across a United Kingdom National Health Service (NHS) Health Authority (HA). We suggest that the evidence based medicine (EBM) movement underpinning such attempts is premised upon a highly rationalistic conception of change. Here the generation and implementation of research findings into clinical practice is understood as movement between discrete entities. Drawing upon poststructural philosophy, social studies of science and technology, social anthropology, and gender studies, we challenge such linear perspectives through a more immanent alternative. We conceive of change as movement within indeterminate or ambiguous relationships. We then proceed to discuss the implications of this modality for the management of clinical behaviour change. PMID- 9877341 TI - Political repression and its psychological effects on Honduran children. AB - This paper, based on an exploratory study in Honduras, examines the psychological effects of political disappearance and assassination on surviving child family members. There are few, if any, comparative field studies of non-immigrant, non refugee or non-clinical populations of children with forcibly disappeared and assassinated parents. In Latin America, violent state power has been exercised in a number of ways, including physical intimidation, jailing, murder and "disappearance". This research compares 16 Honduran children who have experienced the loss of a parent through forced disappearance with 11 Honduran children who suffered a similar loss from a political assassination. Surviving parents or caretakers of these children were also interviewed. Differences between children of the disappeared and assassinated were analyzed using a Rorschach object relations scale, post-traumatic stress and symptom and behavior scales and interviews with the children and the surviving parents. Both groups of children have symptoms of traumatic stress, depression, anxiety and aggressive feelings. Children of both groups remain symptomatic for many years after their loss and children with disappeared parents present more unconscious emotional disturbance in the Rorschach object relations scale than children with assassinated parents. The lack of physical, emotional and legal resolution in forced disappearance appears to impede mourning and a return to age-appropriate concerns. PMID- 9877344 TI - The evolution of professional identity: the case of osteopathic medicine. AB - The osteopathic medical profession was founded in the late 19th century and has become an accepted part of the medical establishment in the United States. Throughout its history, the osteopathic medical profession has attempted to define itself in a way that differentiates osteopathy from other alternative therapies and situates the profession as responsive to the changing health care needs of the American public. This article examines identity within the osteopathic profession by examining the ways in which the profession has created, maintained, and changed its identity in its over century-long existence. The case analysis presented here involves the examination of identity statements culled from several osteopathic data sources. The identity statements represent four specific time periods within the osteopathic profession: the founding statements of A. T. Still, statements from 1915 through 1935 when the scope of osteopathic identity was expanding, statements from 1954 through 1974 in which the osteopathic profession dealt with internal and external threats in developing a "separate but equal" identity, and recent statements from a osteopathic student web site that illustrate current and future views of osteopathic identity. The results of this case analysis highlight the role of the social environment in establishing and changing professional identity, the importance of an occupational founder in shaping the articulation of identity, and the tension between identity and practice within the osteopathic medical profession. PMID- 9877345 TI - Promoting health, promoting women: the construction of female and professional identities in the discourse of community health workers. AB - Community health worker (CHW) programs are implemented in the third world and among racial minorities in the U.S. by public health professionals with the goal of improving people's access to basic health services. There is a shared view that women's roles as mothers make them effective CHWs because most health practices are located within the realm of the family. The objective of this paper is to inquire how and what concepts of woman are constructed and promoted in CHW programs. Viewing CHW as a discourse, I examine literature on CHWs using a critical feminist perspective and insights from narrative and rhetorical analyses. I argue that CHW positions women living in the third world and non white Hispanic women in the U.S. as the "other" woman. The natural attributes of this other woman include mother, care giver, oppressed, child-like, and victim of patriarchy, religion, poverty, and diseases. These attributes are used to define categories of the female such as "the third world woman" and "Hispanic woman". These categories, in turn, define two unnamed opposite categories: "the first world woman" and "the public health professional". I conclude that CHW is a colonizing discourse and that public health professionals and feminists need to practice reflexivity. PMID- 9877346 TI - Psychosocial stressors and well-being in health care workers. The impact of an intervention program. AB - There have been few prospective studies of the impact of workplace interventions on employee and organizational well-being within health care settings. This study was conducted at a large regional hospital in Sweden in 1994 with a follow-up in 1995. Effects of a structured organizational and staff intervention program on perceived psychosocial work quality, supporting resources and self-reported health and well-being were evaluated. Based on department-specific results from the baseline assessment in 1994, each department formulated their own improvement goals. They also made their own decisions on relevant improvement activities. Since there was no formal reference group in this study, departments with high and low rating levels, respectively, with regard to intervention activities were compared. Despite an overall worsening in most of the measures most likely due to a notice of 20% staff reduction prior to the follow-up assessment, the intervention appeared to have attenuated negative changes in the high as compared with the low activity group. Manager-rated impact of the program as well as positive staff attitudes and staff involvement in the enhancement process were identified as important determinants for more favourable changes. The study points out the relevance of structured workplace interventions for organizational and employee well-being especially in times of cut-backs and organizational turmoil. Department-specific factors will determine the impact of such programs. The study indicates that the psychosocial impact of personnel cut-backs in health care may be attenuated through management initiatives. PMID- 9877347 TI - Gender differences in accessing cardiac surgery across England: a cross-sectional analysis of the health survey for England. AB - OBJECTIVE: To examine gender differences in access to cardiac surgery in a nationally representative sample. DESIGN: Nationwide cross sectional household based survey (Health Survey for England). SETTING: Private households in England around 1993 and 1994. SUBJECTS: 1708 subjects reporting a history of either doctor diagnosed angina or heart attack from a stratified random sample of 32 378 people aged 16 and above. OUTCOME MEASURE: The proportion reporting having had cardiac surgery or on a waiting list. RESULTS: 13.5% reported previous (n = 206) or pending (n = 25) cardiac surgery. Men were more likely than women to have had or to be waiting for cardiac surgery (19.1% of men versus 6.8% of women, chi2 54.7, P<0.001). This finding was consistent regardless of age group and across three regional areas. The unadjusted odds ratio for cardiac surgery for men versus women was 3.3 (95% Cl 2.3, 4.5, P<0.001) and was only slightly attenuated to 2.8 (95% CI 1.9, 4.0. P<0.001), after adjustment for other factors. The gender difference remained even when analysis was restricted to subjects reporting a previous heart attack, and after statistical adjustment for disease severity. CONCLUSION: Women are less likely than men to receive cardiac surgery across all age groups and regional areas. These results include private operations and adjust for individual behavioural data. Neither disease severity or co-morbidity explains these discrepancies. Further studies are required to determine why this inequality occurs and how it can be addressed. PMID- 9877348 TI - Why do nurses abuse patients? Reflections from South African obstetric services. AB - Nurse-patient relationships are a substantially neglected area of empirical research, the more so in developing than developed countries. Although nursing discourse usually emphasises "caring", nursing practice is often quite different and may be more strongly characterised by humiliation of patients and physical abuse. This paper explores the question: why do nurses abuse patients, through presentation and discussion of findings of research on health seeking practices in one part of the South African maternity services. The research was qualitative and based on 103 minimally structured in-depth individual interviews and four group discussions held with patients and staff in the services. Many of the patients reported clinical neglect, verbal and physical abuse from nursing staff which was at times reactive, and at others, ritualised, in nature. Although they explained nurses' treatment of them in terms of a few 'rotten apples in the barrel', analysis of the data revealed a complex interplay of concerns including organisational issues. professional insecurities, perceived need to assert "control" over the environment and sanctioning of the use of coercive and punitive measures to do so, and an underpinning ideology of patient inferiority. The findings suggest that the nurses were engaged in a continuous struggle to assert their professional and middle class identity and in the process deployed violence against patients as a means of creating social distance and maintaining fantasies of identity and power. The deployment of violence became commonplace because of the lack of local accountability of services and lack of action taken by managers and higher levels of the profession against nurses who abuse patients. It also became established as "normal" in nursing practice because of a lack of powerful competing ideologies of patient care and nursing ethics. The paper concludes by discussing avenues for intervention to improve staff-patient relationships. PMID- 9877349 TI - Describing the "white" ethnic group and its composition in medical research. AB - The routine use in medical research of an ostensibly homogeneous "white" category in ethnic group classifications has meant that white minorities, such as the Irish, Turks and Cypriots, have remained hidden, even though such groups are subject to discrimination and disadvantage common to other minority groups. The terms "white" and "Caucasian" are frequently and increasingly employed in the scientific literature in spite of widespread concern about the medicalization of race. Moreover, in Great Britain ethnic monitoring of hospital inpatients has revealed negligible interest in utilising codes that subdivide the white group. Yet recent research has shown, for example, substantially elevated age standardised limiting long-term illness rates in the first generation Irish and excess mortality in the second generation living in Britain. The health needs of these white minorities can only properly be identified through the availability of census denominator data of the kind now collected in the U.S. and Canadian decennial census questions on ethnic origin. The opportunity for government to make such provision in the forthcoming Great Britain 2001 Census should be seized whilst it is still available and recommendations for subdividing the "white" group are made.